PMID- 10665265 TI - Neuropsychiatry of Huntington's disease and other basal ganglia disorders. AB - Degenerative diseases of the basal ganglia, such as Huntington's disease (HD), Parkinson's disease, and Wilson's disease, are characterized by motor, cognitive, and psychiatric manifestations. HD, in particular, can be considered a paradigmatic neuropsychiatric disorder that has all three components of the "Triadic Syndromes": dyskinesia, dementia, and depression. The authors examine the phenomenology, prevalence, and management of psychiatric disturbances occurring in diseases of the basal ganglia. They address psychiatric conditions such as depression, mania, psychosis, obsessive-compulsive disorders, aggression, irritability, apathy, sexual disorders, and delirium, discussing subtleties of diagnosis, and making reference to more unusual disorders of the basal ganglia, such as postencephalitic parkinsonism and Fahr's disease. PMID- 10665266 TI - The psychiatric perspectives of epilepsy. AB - Psychiatric conditions occur frequently in epilepsy, and their manifestations are diverse. Evaluation and management require knowledge of disease processes relevant to epilepsy and to psychiatry, as well as the role of other factors that affect the expression of psychiatric illnesses: behaviors, temperament, cognition, and life events. This article describes a comprehensive approach for addressing psychiatric issues in epilepsy patients. PMID- 10665267 TI - Interventions to improve provider diagnosis and treatment of mental disorders in primary care. A critical review of the literature. AB - The authors conducted a critical review of the literature on interventions to improve provider recognition and management of mental disorders in primary care, searching the MEDLINE database for relevant articles published from 1966 through May 1998 and finding 48 usable controlled studies (27 randomized controlled trials and 21 quasi-experimental studies). Improved diagnosis of mental disorders was reported in 18 of 23 (78%) of the studies examining this outcome and improved treatment in 14 of 20 studies (70%); clinical improvement in psychiatric symptoms or functional status was documented in 4 of 11 and 4 of 8 (36% and 50%, respectively). Considerable study heterogeneity precluded subjecting the literature synthesis to a formal meta-analysis of pooled results; the authors were therefore unable to demonstrate an association between efficacy of an intervention and any specific variables. A variety of interventions and further research may be effective in improving the recognition and management of mental disorders in primary care. PMID- 10665268 TI - Sharing mental health care. Training psychiatry residents to work with primary care physicians. AB - Overcoming problems in communication between psychiatry and primary care requires new models of collaboration. Their success will depend upon the ability of participants to work productively with each other, which will require psychiatry residency programs to offer appropriate preparation for future graduates in working with primary care physicians. This article, based on the training at McMaster University in Hamilton, Ontario, describes a brief curriculum for training psychiatry residents to work effectively with primary care physicians that can be easily integrated with current training rotations and looks at adjustments academic departments need to make to support such programs. PMID- 10665269 TI - Psychiatric complications of Ma-huang. PMID- 10665270 TI - QT interval prolongation associated with quetiapine (Seroquel) overdose. PMID- 10665271 TI - Psychiatric effects of anabolic steroids after burn injuries. PMID- 10665272 TI - Low-dose risperidone for the irritable medically ill patient. PMID- 10665273 TI - A psychotic manic state induced by an herbal preparation. PMID- 10665274 TI - Panic disorder with agoraphobia in reaction to gastroenteritis. PMID- 10665275 TI - [Clinical variants of glaucoma in patients with acquired high myopia]. AB - Examinations of 88 glaucoma patients with high myopia showed three variants of glaucoma: 1) mixed form with combination of nuclear cataract and lenticular myopia, 2) open-angle form with ischemic background cerebral circulation (ocular circulation deficiency combined with a decrease in linear velocity of the bloodflow in the intracranial collateral cerebral circulation: to 21.8% in the anterior cerebral artery, to 26.1% in the middle cerebral artery, and to cerebral artery (by 8.3%) and in the orbital artery (by 4.2%). Clinical values indicate impeded venous outflow. Differential diagnostic signs of glaucoma in high myopia are defined. PMID- 10665276 TI - [Endoscopic laser coagulation of ciliary processes in patients with severe uncompensated glaucoma]. AB - Endoscopic laser coagulation was used in 85 patients with severe uncompensated glaucoma. A manifest hypotensive effect (decrease of intraocular pressure from 44.21 +/- 3.17 to 23.12 +/- 4.58 mm Hg 1 year after the operation) and analgesic effect were attained. As a result of treatment, intraocular pressure normalized in 88.2% patients and the eyeball was preserved in 98.8% patients subjected to surgery. PMID- 10665277 TI - [Clinical and pathogenetic features of neurotrophic corneal disorders in diabetes]. AB - The corneal neurotrophic status was evaluated in 110 patients (220 eyes) with diabetes mellitus and in 20 normal controls (40 eyes). Corneal abnormalities (gerontoxon, limb vascularization, punctate keratopathy, endothelial dystrophy, relapsing erosion, corneal ulcers) were detected in 73.6% diabetics. Corneal changes depended on the type, duration, and compensation of diabetes mellitus. Based on the diagnostic markers (threshold sensitivity of the cornea, microcirculatory status, bulbar conjunctiva status, level of lacrimal production, cellular composition of the limb smears impressions, lacrimal levels of catecholamines, hydroperoxides, and lipids), three types of diabetic neuropathy of the cornea were distinguished: vegetative, sensory, and vegetosensory. Corneal changes were associated with signs of diabetic neuropathy in 55.6% cases and with autonomic nervous disorders of the cardiovascular system in 46.9% cases, which evidences a generalized involvement. The detected neurotrophic features of the cornea should be borne in mind when choosing pathogenetic therapy for diabetics with corneal complications and preparing the patients to ocular surgery. PMID- 10665278 TI - [Objective method for epithelialization and corneal transparency monitoring after excimer laser keratectomy]. AB - The area of corneal epithelialization was evaluated and densitometry of corneal surface carried out by computer analysis of images in order to detect the relationship between epithelialization and formation of corneal fleur in patients with delayed healing after photorefraction keratectomy for hypermetropia. An arbitrary parameter: corneal epithelialization rate, a mathematical difference between the area of corneal defect every next day of observation in comparison with the day before, expressed in mm2/day, is proposed for evaluating the efficacy of various drugs used to stimulate corneal defect epithelialization. Computer analysis of images helps objectively evaluate epithelialization and corneal transparency after eximer laser keratectomy. The method is diagnostically and prognostically valuable, particularly in cases with delayed epithelialization and stubborn epithelial defects of the cornea. PMID- 10665279 TI - [New stimulants of corneal reparative regeneration]. AB - The efficacy of corneregel, a drug containing pantothenic acid, a component of coenzyme A, in healing of corneal wounds has been evaluated. The study was carried out on 19 rabbits (38 eyes) with standard corneal defect made with a 5-mm trephine for lamellar transplantation of the cornea, divided into 2 groups: 1) instillations of corneregel (10 eyes) and 0.25% levomycetin solution (10 eyes) and 2) 20% solcoseryl gel (9 eyes) and 0.25% levomycetin (9 eyes). Time course of changes were evaluated by biomicroscopy (fluorescent test), histologically (hematoxylin-eosin staining), and immunohistochemically after 1, 2, 4, 7, 30, and 90 days. Proliferative activity was studied by expression of the proliferating cell nuclear antigen and the migration capacity of cells by expression of alpha smooth muscle actin. The terms of epithelialization were as follows: corneregel 10 +/- 7 h, 20% solcoseryl gel 108 +/- 10 h, levomycetin 124 +/- 6.93 h. Earlier epithelialization in the corneregel group was apparently due to increased expression of alpha-smooth muscle actin and increase in the cell migration capacity. Hence, corneregel is recommended for practical use as a stimulant of reparative regeneration of the cornea. PMID- 10665280 TI - [Combined revascularizing method for treating central atherosclerotic chorioretinopathy]. AB - The authors present a new surgical method for treatment of central atherosclerotic chorioretinopathies, consisting of choroid revascularization followed by crossing of the surface temporal artery. Combined revascularizing method improved local ocular hemodynamics and the visual function in 64% patients with pseudotumorous condition and in 81% patients with the "dry" form, stabilizing the process for up to 12 months (period of observation). Morphological analysis showed that after choroid revascularization, a full-value functioning vascular network forms at the interface between the vascular membrane and implanted tissues, and therefore crossing of the surface temporal artery should be performed no earlier than 3 months after choroid revascularization. PMID- 10665281 TI - [Peripheral chorioretinal dystrophy and astigmatism of myopic eyes: statistical evaluation, relationship and clinical significance]. AB - A clinically significant relationship between the myopic eye astigmatism and manifestation of peripheral chorioretinal dystrophies in them in young patients is described. A simple and effective method for predicting peripheral vascular chorioretinal dystrophies in myopic patients is proposed, based on the presence and severity of astigmatism. The incidence of various dystrophies in spherical and aspherical myopia is 32 and 78%, respectively. Dystrophies fraught with retinal detachment occur three times more often in astigmatic eyes, which are characterized by a more flat lens. PMID- 10665282 TI - [Morphology of conjunctival progressive nevus]. AB - Structural features of 59 progressive nevuses of the conjunctiva were studied. The proliferation of the epithelium and melanocytes is partially compensated by spontaneous regression of the nevus structures. The growth of a nevus is structurally similar to tumor growth, but the nevuses lack the melanocyte dysplasia, the main sign of malignant degeneration. The immune reactions are involved in the tissue restructuring of the growing nevus. Permanent foci of photoelastosis reflect the significance of ultraviolet exposure as a factor of risk of the nevuses progress. PMID- 10665283 TI - [Radionuclide imaging of lacrimal duct in health and disease]. AB - Dacryoscintigraphy is a perspective method for examination of the lacrimal system function. It has many advantages over other methods (functional tests, roentgenography, and computer-aided tomography) and notably improves the diagnostic potentialities of examination of the lacrimal function. The technique and results of the method are discussed. PMID- 10665284 TI - [Prediction of infectious complications of eye penetrating wounds by R protein concentration in lacrimal fluid]. AB - Analysis of the lacrimal fluid of normal subjects and patients after cataract extraction and with penetrating wounds of the eyeball showed fluctuations of the R protein concentration, which depended on the type of the injury and its complications. The authors propose using measurements of R protein in the lacrimal fluid as a prognostic criterion predicting the development of infectious complications in penetrating wounds of the eyeball. PMID- 10665285 TI - Electroretinographic characteristics of proliferative vitreoretinopathy in rhegmatogenous detachment of retina. AB - Thirty-five patients with types A, B, C2, C3, and D1 proliferative vitreoretinopathy (PVR) were followed up for 8-14 months after surgical treatment of rhegmatogenic amotio retinae. Hanz-Feld electroretinogram (ERG), rhythmic ERG to low and high-frequency stimulation, and macular ERG were recorded. Before surgery all patients presented with specific signs of changes in the retinal electrogenesis, characteristic of rhegmatogenic amotio retinae with PVR of different types. Comparative analysis of ERG and rhythmic ERG alteration showed a more expressed reaction of Mueller's glial cells in patients with types A-C2 PVR and of second order neurons in the internal nuclear layer of the retina in those with types C3 and D1 PVR. Characteristic changes in the retinal functional activity were detected, possessing a prognostic significance. According to electrophysiological studies, the decisive factor for predicting the course of recovery after surgical treatment of retinal detachment is the preoperative function of retinal neurons. A drastic depression of low-frequency rhythmic ERG is the most unfavorable sign in prognosis of the postoperative time course of visual functions. The least favorable course of recovery after the operation is typical of amotio retinae with type C3 PVR. Recovery of amplitudes of different biopotentials and functional horizontal bonds at different levels of the retinal structure is different. PMID- 10665286 TI - [Parameters of blood proteinase inhibitor balance and hyperlipoproteinemia in patients with proliferative diabetic retinopathy with vitreous hemorrhage]. AB - The parameters of proteinase-inhibitor balance and lipid metabolism were studied in the blood of patients with proliferative diabetic retinopathy. Metabolic disorders in patients with proliferative diabetic retinopathy with hemophthalmia are characterized by a notable increase in the activities of trypsin-like enzymes and alpha 2-macroglobulin level and normal activity of alpha 1-antitrypsin and shifted lipid metabolism parameters characteristic of types IIb and IV hyperlipoproteinemia. These data indicate a necessity of monitoring lipid metabolism and proteinase-inhibitor balance in patients with diabetic retinopathy in order to detect subjects at a high risk of hemophthalmia and timely start drug correction for preventing this complication. PMID- 10665287 TI - [Relationship between perinatal pathology and refractogenesis, incidence and type of ocular diseases in children]. AB - Three-year-old children with a history of perinatal diseases differed from healthy age-matched children by a higher incidence of ocular diseases (78.9% vs. 21.6% in the control, p < 0.001). These children often presented with severe visual disorders: partial atrophy and hypoplasia of ocular nerves (7.2%), congenital abnormalities in the eyeball membranes (5.2%), retinopathy neonatorum (5.2%), cortical blindness (3.1%), oculomotor disorders (20.8%), and congenital deformations of the eyelids (19.7%). Disorders of refractogenesis in these children presented as a higher incidence of myopia (19.8% vs. 3.8% in the control, p < 0.001) and a shift of the percentage of refraction abnormalities towards myopia. Therefore, all children with a history of perinatal disease should be referred to a group at a high risk of ocular disease. PMID- 10665288 TI - [Interferon status of patients with uveal melanoma]. AB - Interferon status of patients with uveal melanoma was studied at different stages of the disease. All parameters of alpha- and gamma-interferon production were decreased both in vitro and in vivo. The T2-3 stages were associated with an imbalance of the production of two interferon types: augmenting deficiency of alpha-interferon with a tendency to a higher production of gamma-interferon by leukocytes in vitro. Generalization of T3-4N0Mhep melanoma was characterized by hyperproduction of gamma-interferon and total serum interferon. These data indicate a pathogenetic significance of disorders in interferon production in uveal melanoma and may be used for the disease prognosis. PMID- 10665289 TI - [Therapy of fundus oculi vascular pathology by solcoseryl]. AB - Long (for more than 17 years) therapy of 2331 patients (3122 eyes) with vascular conditions of the fundus oculi by a retinotropic drug solcoseryl showed its high efficacy as a monotherapy and in complex with other traditional and symptomatic treatments. Solcoseryl improved the visual function and hemodynamics of retinal vessels, promoted a more stable and longer stabilization of the treatment results, and accelerated the rehabilitation of patients. PMID- 10665290 TI - [Effects some drugs of re-epithelialization in the early postoperative period after photorefraction keratectomy]. AB - Terms of re-epithelialization, severity of the painful syndrome, intensity of corneal "crepe" (opacity) are assessed in myopic patients treated by maxitrol, eubetal, colbiocin ointments and maxitrol eyedrops in the early postoperative period after photorefraction keratectomy. The crepe intensity was assessed routinely according to a clinical score: 0) transparent cornea, 1) trace crepe; 2) moderate crepe; and 3) intensive crepe. Biomicroscopy on day 4 after photorefraction keratectomy showed complete epithelialization in 91.7% patients after colbiocin ointment, in 91.% after maxitrol eyedrops, 87% after eubetal ointment, and 82.6% after maxitrol ointment. The least corneal opacity (0 and 0 1) was observed after eubetal ointment and maxitrol eyedrops. The mean score for pain was virtually the same in all groups; in the colbiocin ointment group more patients complained of pain for more than 24 h in comparison with other groups. PMID- 10665291 TI - [Optic nerve hypoplasia]. PMID- 10665292 TI - [Endocrine ophthalmopathy]. PMID- 10665293 TI - Detailed diagnoses and procedures, National Hospital Discharge Survey, 1997. AB - OBJECTIVES: This report presents national estimates of the use of non-Federal short-stay hospitals in the United States during 1997. Estimates of first-listed diagnoses, all-listed diagnoses, days of care for first-listed diagnoses, and all listed procedures are shown by sex and age of patient and geographic region of hospital. METHODS: The estimates are based on data collected through the National Hospital Discharge Survey for 1997. The survey has been conducted annually by the National Center for Health Statistics since 1965. In 1997 data were collected for approximately 300,000 discharges from 474 non-Federal short-stay hospitals. Diagnoses and procedures are presented according to their code number in the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM). PMID- 10665294 TI - [The power of numbers]. AB - The round figure for the current year has stirred people's minds in anticipation. Numbers have acquired great significance also in today's medical science. The Paris physician Pierre Charles Alexandre Louis (1787-1872) is considered the founding father of the numerical method in medicine. At first the principle of aggregating data from different individuals aroused much resistance and even disgust: Claude Bernard was a leading figure among those who warned that one will never find a mean in nature, and that grouping findings together obscures the true relationship between biological phenomena. True enough, statistical significance is not a characteristic of nature itself. Significant differences or risk reductions do not necessarily imply clinical relevance, and results obtained in a group of patients are rarely applicable to an individual patient in the consultation room. Likewise, the health of a human being cannot be captured in biochemical, radiological or other technical measures, nor in disease-specific scales that reduce well-being to one or two digits. The editors of this journal will remain keen on publishing numerical studies that contribute to evidence based medicine, but at the same time they will continue to foster the art of reporting illness from the point of view of the sick person. PMID- 10665295 TI - [1999, the year of fired editors-in-chief]. AB - In the past year three editors of medical scientific journals stepped back, a remarkable fact for a relatively quiet profession. G. Lundberg was dismissed. J. Kassirer was forced out and F. Meijman resigned. There are differences and similarities in these events. Lundberg of The Journal of the American Medical Association (JAMA) was dismissed because of his interference with regular points of view of the board of the American Medical Association, whereas Kassirer of The New England Journal of Medicine was forced out mainly because of his refusal to allow use of the name and reputation of his journal for other medical and lay publications by the Massachusetts Medical Association; Meijman resigned because he disagreed with the changes in the scientific contents of Huisarts en Wetenschap (General Practitioner and Medical Science) proposed by the board of the general practitioners' association that owns the journal. The similarity is that editorial freedom appears to be a relative concept. Owners, whether societies or publishers, can fire displeasing editors or force them to leave. The consequences could be harmful, to the owners, but especially to the journals, because of loss of impact, credibility and trust by authors, reviewers and readers. The only basis for a constructive co-operation between owners and editors is mutual trust. Immediate firing of editors is not possible at the Nederlands Tijdschrift voor Geneeskunde (Dutch Journal of Medicine). The association that owns the journal consists of (former) members of the editorial board. A committee of the association assesses every year the editorial decisions and policies retrospectively. Subsequently the editors may be reappointed for another year by a vote of the general assembly of the association. If they are not reappointed, an administrative procedure will be initiated to terminate their contract. PMID- 10665296 TI - [Epilepsy, disturbances of behavior and consciousness in presence of normal thyroxine levels: still, consider the thyroid gland]. AB - Three patients, one man aged 51 years, and two women aged 49 and 52 years, had severe fluctuating and progressive neurological and psychiatric symptoms. All three had normal thyroxine levels but elevated thyroid stimulating hormone levels and positive thyroid antibodies. Based on clinical, laboratory, MRI and EEG findings they were eventually diagnosed with Hashimoto's encephalopathy, associated with Hashimoto thyroiditis. Treatment with prednisone in addition to thyroxine suppletion resulted in a remarkable remission of their neuropsychiatric symptoms. The disease is probably under-recognized. PMID- 10665297 TI - [Laparoscopic inguinal hernia operation]. AB - After the introduction of laparoscopic operations for repair of inguinal hernias, circa 1990, randomized comparative studies showed that this method causes less postoperative pain and requires a shorter recovery period than conventional operations. The duration of follow-up is still too short for comparison of proportions of recurrences. The costs of laparoscopic interventions are higher for the hospital (among other things because of the use of disposable instruments) and lower for society (because of shorter absenteeism). Implementation of the intervention by allotment of a higher fee to hospitals has not been done, because meanwhile a third method--conventional surgery with implantation of a prosthetic mesh in ambulatory surgery--has become possible. A meta-analysis based on European registration of all treated patients will once more have to demonstrate what method of treatment is to be preferred. PMID- 10665298 TI - [Phenylketonuria: a children's disease in adulthood]. AB - The prognosis for patients with phenylketonuria (PKU) has improved greatly with early institution of treatment after birth. It was assumed that the diet could be terminated after adolescence but there are strong indications that hyperphenylalaninaemia can have detrimental effects in adult patients. Hyperphenylalaninaemia can cause reversible white matter abnormalities, and is also associated with psychiatric illness, which improves with lowering of the plasma phenylalanine level. Even optimally treated patients generally have a decreased performance with neuropsychological tests compared with subjects without PKU. Elevation of the plasma phenylalanine level leads to worsening of neuropsychological performance, lowering of the level leads to improved performance. Strict metabolic control is necessary during pregnancy in women with PKU in view of the increasing frequency of congenital abnormalities with increasing phenylalanine level. The complexity and demanding nature of the diet treatment make specialised facilities for optimal medical and paramedical care mandatory. PMID- 10665299 TI - [Respiratory syncytial virus infections and preventive options]. AB - Respiratory syncytial virus (RSV) is the most prominent pathogen found in respiratory tract infections in children and the most important cause of bronchiolitis in the first two years of life. In the Netherlands approximately 2000 children are admitted each winter season. A serious course is mostly seen in children younger than 3 months, (ex-)prematures, children with bronchopulmonary dysplasia or congenital cardiac anomalies, children with cystic fibrosis younger then 2 years and children with impaired T cell immunity; such cases not rarely require intensive care. Treatment (fluid, nutrition, bronchodilator agents, corticosteroids, oxygen and ventilation) is usually symptomatic. Antiviral therapy is only indicated in immunodeficient patients. For prevention by passive immunization palivizumab was recently registered in the Netherlands, a monoclonal antibody against RSV that has to be administered intramuscularly from the start of the RSV season (15 mg per kg bodyweight once a month during five months). In a number of large-scale American multicenter studies both the number of hospital admissions related to RSV infection and the mean duration of hospital stay showed a statistically significant reduction in high-risk children who had been treated with palivizumab. Palivizumab appears to be indicated in children from the categories with an increased risk for serious RSV disease. PMID- 10665300 TI - [Low incidence of nosocomial respiratory syncytial virus infections among children younger than 12 months in the Department of Pediatrics, Sophia Children's Hospital at Rotterdam]. AB - OBJECTIVE: To investigate the occurrence of nosocomial respiratory syncytial virus (RSV) infections and to compare their clinical features with those of community-acquired RSV infections. DESIGN: Retrospective. METHOD: Data were collected from the medical records of children younger than 12 months with RSV infection in the Department of Pediatrics of Sophia's Children's Hospital, Rotterdam, the Netherlands, in October-March 1992/'95. The diagnosis of 'RSV infection' was confirmed by a direct immunofluorescent assay and/or a viral culture on materials obtained from nasopharyngeal washes. A nosocomial RSV infection was defined as an infection which occurred more than 5 days after hospital admission for any underlying disease. RESULTS: During the 3 RSV seasons 1260 children were admitted. Of these 34 (2.7%) developed a nosocomial RSV infection. The number of nosocomial RSV infections decreased over the study period. At the department including the outpatient clinic 232 children were seen with a community-acquired RSV infection. Children with a nosocomial infection differed from children with a community-acquired infection only with regard to birth weight (2.5 kg versus 3.0 kg), cough (65% versus 92%) and feeding problems (100% versus 69%). Four children had bronchopulmonary dysplasia and nosocomial RSV infection; these required mechanical ventilation. CONCLUSION: The number of nosocomial RSV infections decreased over 3 years. The severity of nosocomial RSV infections was comparable with that of community-acquired RSV infections. PMID- 10665301 TI - [Man with cherubism]. AB - Cherubism was diagnosed in a male when he was 6 years old. Cherubism is a benign, bilateral, painless lesion. It is commonly located in the mandible but in a minority of patients also in the maxilla. Cherubism is a familial condition with an autosomal dominant pattern of inheritance. At least one member of the family of the patient described had cherubism. The disease becomes manifest during early childhood and progresses until puberty when it spontaneously regresses. In the majority of patients no treatment is required. However, in the patient presented surgical procedures and odontological corrections were necessary. Due to a flare up of the disease at the age of 22 years, the patient was treated with calcitonin for 1 year followed by bisphosphonates. During these treatments the disease symptoms diminished. PMID- 10665302 TI - [Value of a Dutch medical license in Egypt]. AB - Shortly after moving from the Netherlands to Egypt, the author, a physician, started procedures to get his medical certificate recognized. These procedures began at the Egyptian Medical Association, housed in the Dar al-Hikmah ('House of Wisdom'). The Association first required a comparison between the Netherlands and the Egyptian medical education, to be made by the Supreme Council of Universities. There, the differences between the two systems appeared to be fertile soil for confusion and discussions. After nine months, however, and the translation of all relevant papers into Arabic, the Supreme Council declared the Dutch medical education equivalent to the Egyptian. Now only the permission of the Ministry of Health and Population was required. This Ministry inquired at the Ministry of Foreign Affairs whether an Egyptian education would be recognized in the Netherlands, in order for the 'principle of reciprocity' to be applied. The reply of the Dutch Embassy was positive but progress has stopped since. A registration as a physician in the United Kingdom was obtained without problems and in a short time; this registration might be helpful in efforts to resume the process in Egypt. PMID- 10665303 TI - [New diagnostic technology often offers no advantage in the differential diagnosis of acute abdomen]. PMID- 10665304 TI - [Bilateral blood pressure measurement before and after coronary bypass surgery : an absolute necessity]. PMID- 10665305 TI - [Hypertensive crisis: definition, pathophysiology and treatment]. PMID- 10665306 TI - [Autopsies as an important indicator for quality control]. PMID- 10665307 TI - [The "Year 2000 problem"]. PMID- 10665308 TI - [Mechanisms of tachyphylaxis in regional anesthesia of long duration]. AB - Tachyphylaxis to local anesthetics is defined as a decrease in duration, segmental spread or intensity of a regional block despite repeated constant dosages. However, there is disagreement about the incidence of tachyphylaxis. In contrast to tachyphylaxis, pseudotachyphylaxis denotes time dependent variations in pain or circadian changes in the duration of local anesthetic action. Tachyphylaxis appears neither to be linked to structural or pharmacological properties of the local anesthetics nor to the technique or mode of their administration. The mechanisms underlying tachyphylaxis are open to debate and include changes in pharmacokinetics or pharmacodynamics. Considering pharmacokinetics, local edema, an increased epidural protein concentration, changes in local anesthetic distribution in the epidural space or a decrease of perineural pH could result in decreased diffusion of the local anesthetics from the epidural space to their binding sites at the sodium channel. Increased clearance of local anesthetics from the epidural space may be caused both by increased epidural blood flow or increased local metabolism. Considering pharmacodynamics, antagonistic effects of nucleotides or increased sodium concentration, increased afferent input from nociceptors or receptor down regulation of the sodium channels have been implicated. However, none of these theoretical considerations is supported strongly enough by data to explain tachyphylaxis. A new possibility to maintain for a longer time neural blockade is the design of new ultralong-acting local anesthetics. Liposomal formulations of local anesthetics also appear suitable to provide longer lasting regional anesthesia. The recent observation that NMDA-antagonists as well as NO-synthase inhibitors prevent the development of tachyphylaxis suggests involvement of the nitric oxide pathway in the development of tachyphylaxis. Accordingly, NMDA antagonists or NO-synthase-inhibitors may prevent tachyphylaxis. PMID- 10665309 TI - [Anesthesia monitoring: degree of compliance with guidelines in Austria]. AB - PURPOSE: For the first time an evaluation of standard anesthetic monitoring was performed according to the guidelines of the Austrian Society for Anesthesiology, Resuscitation und Intensive Care Medicine (OGARI). METHODS: A questionnaire was delivered to all medical institutions performing anesthesia in Austria. A descriptive statistical evaluation was performed on all returned and completed questionnaires. RESULTS: Generally, there is a high standard in compulsory monitoring and in PACU (actual compliance > 99%/85.8%). Supplemental equipment is required for disconnection alarm and measurement of inspired oxygen concentration (actual compliance: 98.3%/98.9%). Furthermore, measurement for inspired concentration of volatile anesthetics and relaxometry (actual compliance 68.7%/47.3%) has yet to be completed. University departments and regional hospitals have comparable standards (82.2% vs. 79.6%). CONCLUSIONS: For the first time an Austrian-wide evaluation of anesthetic monitoring investigated the compliance with the 1992 recommendations of the Austrian Society of Anesthesiology, Resuscitation and Intensive Care Medicine. The data demonstrate that these recommendations including the anesthetic monitoring equipment have already been implemented to a high degree. PMID- 10665310 TI - [Respiratory and circulatory parameters as indicators of the postoperative analgesic demand in newborns and infants]. AB - AIM OF THE STUDY: Due to immature cognitive functions, infants are unable to communicate their pain perception verbally. To assess postoperative analgesic demand, the anaesthetist has to rely on observational techniques. Generally, pain expression is considered to be a multidimensional phenomenon consisting of physiological, motor-reflex and behavioural patterns. The majority of observational approaches to pain assessment in infants use the behavioural dimension only, regardless of the fact that pain perception might contribute substantially to the stress response. The aim of this study is to evaluate, whether sensitivity and specificity of a behavioural pain scale (CHIPPS [1]) can be improved by adding physiological measures, especially those representing the stress response. PATIENTS AND METHODS: 30 infants aged 0-12 months and scheduled for unilateral herniorrhaphy were studied prospectively. In addition to 9 behavioural items (crying, facial expression, wrinkling of the forehead, motoric restlessness; posture of fingers, arms, legs, toes and torso) the ratio of actual physiological measurements (heart rate, respiratory rate, blood pressure) and their respective preoperative baseline values were recorded by a single observer in 5 minutes intervals during the first hour after recovery from anaesthesia. Maximal efforts were made to achieve valid measures. Factor analysis was performed to determine the dimensionality of the complete item pool. For additional validity testing, receiver operating characteristic curves (ROC) were calculated using the independent opinion of an experienced clinician as an external criterion. Discriminant analysis was performed to assess the accuracy of a combined behavioural and physiological scale. RESULTS: The factor analysis resulted in two independent dimensions: behaviour and cardiocirculatory measurements. The strong intercorrelations of all behavioural items and the fact, that the affective pain experience is expressed by a specific mimic behaviour, suggest the behavioural dimension to be regarded as pain expression. Because of the strictly orthogonal structure of the factor system, the circulatory and the respiratory dimension lack any relationship to pain experience. In addition to these statistical reasons, considerations on practicability disprove blood pressure and respiratory rate as useful pain indicators: Whereas the observer never failed to obtain a behavioural score, only 60% of the blood pressure measures and 80% of the respiratory rates were valid. In contrast, heart rate counts were obtained in over 99% and thus have to be considered as the only practicable physiological measurement in the early postoperative period. Corresponding to the results of the factor analysis, ROC curves suggest that the ability of the heart rate alone to assess pain is not substantially better than a random process, whereas the behavioural scale performs well. In addition the heart rate failed to improve the accuracy of the behavioural scale as shown by the results of a discriminant analysis. CONCLUSIONS: Despite the multidimensional approach and the corresponding multivariate analyses, a unidimensional scale consisting of behavioural items was found to be a valid indicator of an postoperative analgesic demand. Due to the lack of diagnostic properties and difficulties to obtain sound values even under research conditions, physiological measurements like blood pressure, respiratory rate and heart rate are not suitable for the assessment of a postoperative analgesic demand in infants, neither for clinical nor for research purposes. PMID- 10665311 TI - [Temporary adjustable defibrillator electrodes: an alternative method for the treatment of postoperative arrhythmias after heart surgery]. AB - INTRODUCTION: Supraventricular and ventricular tachycardias are common and serious postoperative complications early after cardiac surgery. We introduce a new temporary adjustable defibrillation electrode (TADE) for internal low-energy cardioversion/defibrillation of postoperative atrial and ventricular tachyarrhythmias. METHODS: Evaluation of the new electrode was performed in ten open-chest beagles with a mean weight of 25.5 kg. The electrodes were first positioned on the left and right atrium. Atrial fibrillation (AU) was induced via a bipolar temporary heart wire. Atrial defibrillation thresholds (DFT's) were measured according to a step-down shock protocol (5 J-0.4 J). Thereafter, the electrodes were adjusted and positioned on the right and left ventricle. Ventricular fibrillation (VF) was induced and DFT's were recorded the same way. RESULTS: For termination of AF, mean DFT's were 0.4 +/- 0 J (lowest possible shock level) with a mean shock impedance of 70 +/- 7.6 omega. VF was terminated with a mean DFT of 3 +/- 1.1 J with a mean impedance 56.1 +/- 7.9 omega. Complete transcutaneous removal of the electrodes was possible in all animals without any complications. CONCLUSION: Successful low-energy termination of AF and VF is possible with the tested temporary adjustable electrode. A clinical study is planned for further evaluation. PMID- 10665312 TI - [Blood loss: physiologic adaptation mechanisms and therapeutic replacement]. PMID- 10665313 TI - [Significance of cellular hydration for cellular function]. PMID- 10665314 TI - Production of human albumin by plasma fractionation. PMID- 10665315 TI - Yeast-derived recombinant human albumin (Recombumin). PMID- 10665316 TI - Large scale production of RH-albumin expressed in the milk of transgenic cattle- an economic and technical challenge. PMID- 10665317 TI - [Human albumin--safety and tolerance]. PMID- 10665318 TI - [Natural and artificial colloids in children]. PMID- 10665319 TI - Use and abuse of albumin in clinical practice. PMID- 10665320 TI - [Splenic rupture as a complication of ventilation in the prone position and pneumococcal sepsis as a late complication]. AB - We are reporting the case of a female patient who had to undergo splenectomy after she suffered splenic rupture as a result from "kinetic therapy" during the treatment for pulmonary failure secondary to sepsis. Four years later the patient was again admitted with a clinical picture consistent with sepsis. Two blood cultures were positive for pneumococci confirming the diagnosis of pneumococcal sepsis. This paper discusses the potential risks of kinetic therapy in patients with ARDS. After splenectomy there is increased risk of infection with certain bacteria, funghi, viruses and protozoa. The most common bacterial pathogen is pneumococcus. A polyvalent vaccine is available for prophylaxis. Although penicillin G is still commonly used as an antibiotic therapy for pneumococcal infection, increased resistance of pathogens to penicillin must be anticipated. Alternative antibiotic regimens are demonstrated. PMID- 10665321 TI - [Quality of research in in university anesthesiology departments in Germany]. PMID- 10665322 TI - [MAPA (Ambulatory Measure of Blood Pressure), non-dippers, or which hypertensive patients to treat?]. PMID- 10665323 TI - [Postoperative infections after heart surgery under extracorporeal circulation]. AB - Postoperative infection is still an important cause of mortality and morbidity after cardiac surgery. The aim of this study was to assess its incidence and causes in order to optimise treatment. Between January 1996 and December 1997, 1,000 consecutive patients (253 women and 747 men) were operated for cardiac aortic pathology under cardiopulmonary bypass. The mean age was 66 +/- 11 years. The initial pathology was coronary artery disease (N = 663), valvular heart disease (N = 193), an association of the two (N = 94), thoracic aortic pathology (N = 38) or other pathologies (N = 12). The global postoperative infection rate was 4.9% (N = 49). The incidence of sternal and/or mediastinal infections was 0.7%, of bronchopneumonia 0.9%, urinary infection 2.1%, and septicaemia 1.7%. Nine patients died of the consequences of an infection. The hospital stay was significantly longer in infected patients, irrespective of the site of infection. Statistical analysis of the whole population did not show any predictive factor related to the preoperative clinical status of the patients. The only predictive factor demonstrated was the day on which surgery was performed: the infection rate in patients operated during the first 4 days of the week was 2.2% compared with 7.3% for the patients operated during the last 3 days (p = 0.004, odds ratio (OR) = 3.57). In those patients who had an urinary infection, the two identified risk factors were the female gender (p = 0.006, OR = 3.34) and an operation performed at the end of the week (p = 0.017, OR = 3.77). In patients with sternal and medistinal infections, the only identified predictive factor was combined coronary artery and valvular surgery (p = 0.009, OR = 7.43). With respect to pulmonary infections, the only predictive factor was recent preoperative myocardial infarction (< 1 month) (p = 0.004, OR = 7.5). Finally, no predictive risk factors were identified in those patients who developed septicaemia. In conclusion, this study showed that postoperative infection remains a serious complication of cardiac surgery. The prevention of these complications should be a priority for quality health care. PMID- 10665324 TI - [AS heterozygote hemoglobinopathy and coronary risk]. AB - There have been several reports of vaso-occlusive events and sudden death in subjects with sickle cell trait. However, the precise mechanism underlying these episodes remains unclear. The clinical observations have been supported by in vitro studies in which haemoglobin AS (Hb AS) red cells showed abnormalities of their filterability, probably related to gelling or polymerisation of the Hb AS. These in vitro studies and reports in the literature of sickle-cell hearts led the authors to investigate the possible association between AS subject and coronary risk. The results of coronary angiography in 9 patients with Hb AS, paired with respect to the usual cardiovascular risk factors, were compared with those of 18 AA subjects. The number of patients who underwent coronary bypass surgery for three-vessel disease was much greater in the AS subjects. However, the difference was not statistically significant. This tendency of AS subjects to develop thrombosis and coronary artery disease requires further study with larger numbers of patients. PMID- 10665325 TI - [Implantable ventricular defibrillator. Systematic induction of ventricular arrhythmia is not useful except for implantation]. AB - Checking the efficacy of a defibrillator after its implantation is current practice. This control usually entails induction of ventricular fibrillation (VF). The aim of this study was to assess the utility of this practice in patients with an endovascular system of electrodes. Implantation was acquired when a margin of security of 10 days or more had been obtained. During the postoperative test, the choice of energy was that which reduced VF at implantation. Of the 84 patients included in this study, 59 were implanted with an endovascular electrode system alone and the other 25 patients had endovascular electrodes associated with a subcutaneous patch electrode. The average time from implantation to the postoperative test was 31 +/- 12 days. Arrhythmia sensing was normal and reduction by the defibrillator was observed in all patients. The average energy of the electric shocks was not significantly different to that observed at implantation (18.6 +/- 3.6 J and 19.3 +/- 4.7 J). Fifty patients had antiarrhythmic drugs (amiodarone = 43) at implantation, and 32 at the time of the postoperative test (amiodarone = 17). This study showed that the postoperative test with induction of VF was normal in all patients. Therefore, the authors propose that follow-up should consist of consultation including interrogation of the defibrillator and a chest X-ray to check the position of the electrodes. If one of these tests is abnormal or if the perioperative threshold of defibrillation does not provide an adequate margin of safety, induction of a ventricular arrhythmia is necessary to check the function of the system. PMID- 10665326 TI - [Hypertrophic obstructive cardiomyopathy and double-chamber pacing. Long-term results in a consecutive series of 22 patients]. AB - The authors report their experience with dual-chamber pacing in hypertrophy obstructive cardiomyopathy. 22 patients (14 women and 8 men) mean age 60 +/- 13 years were implanted between 1992 and 1998. The criteria for pace-maker implantation were the presence of severe symptoms related with hypertrophy obstructive cardiomyopathy (dyspnea, angina, syncope) and left ventricular outflow tract gradient at mean 30 mmHg. Before pacing, all patients received a medical therapy which included beta-blockers or calcium inhibitors. This treatment was considered as ineffective or responsible of side effects. Patients were followed-up at mean 35.1 +/- 20.3 months. During this period, symptoms improved (mean NYHA class 2.7 +/- 0.5 before pacing vs 1.4 +/- 0.5 after pacing) and left ventricular outflow tract lowered from 95.4 +/- 40.8 to 39.3 +/- 20.5 at 6 months. 34.3 +/- 23.4 at one year and 26.5 +/- 21 at the end of follow-up. Seven patients had RF ablation of atrio-ventricular junction for paroxysmal atrial fibrillation or for lack of hemodynamic improvement with pacing. This procedure permits a significative lowering of gradient and a better ventricular filling. In conclusion, dual-chamber pacing is effective for treatment of hypertrophy obstructive cardiomyopathy when medical therapy is ineffective or bad tolerated at condition of: perfect pacing with permanent ventricular capture and optimal AV delay; RF ablation of AV junction in one third of cases; medical therapy systematically associated in all patients. PMID- 10665327 TI - [Abnormalities in the circadian rhythm of arterial blood pressure. Physiopathological mechanism and clinical implications]. AB - A wide range of definitions is used to distinguish the hypertensives with a blunted circadian pattern of blood pressure, labeled as "non dippers", from those presenting with a normal night blood pressure fall, designated as "dippers". Consequently the prevalence of non dipping phenomenon is quite uncertain: from 6 to 40% of the hypertensive subjects. The mechanisms of night blood pressure fall remain unclear, but the involvement of autonomic nervous system turns out to be partly demonstrated. From a clinical point of view, non dippers and dippers characteristics are not obviously different, but the prognosis of non dipper patients appears to be rather poor, with a more frequent target organ damage and a higher rate of cardiovascular events, as compared to dippers, even in a general population. In order to improve the identification of non dippers, an ambulatory blood pressure monitoring should be performed in patients with autonomic nervous system impairment and in subjects with target organ damage more severe than expected from office or home blood pressure measurements. PMID- 10665328 TI - Cardiac metabolism in ischemic heart disease. AB - Myocardial ischemia is the metabolic consequence of an inadequate blood supply to the myocardium. How does the myocardium survive a severe insult? Ischemia, perhaps acting through hypoxia, is able to induce a series of cellular signals that lead to protective genetic reprogramming. Metabolic self-protection includes the new ischemic syndromes: stunning, hibernation and preconditioning. In every case it should be considered that ischemia is basically a metabolic problem, usually caused by coronary artery disease, stemming from lack of oxygen and blood flow. In principle, besides revascularization, metabolic therapy should be considered. In the past, metabolic therapies for effort angina have often been ignored. A current hypothesis is that the ischemic myocardium benefits from a switch from fatty acid to glucose metabolism. Two examples are (1) the hemodynamically neutral antianginal agent, trimetazidine, and (2) intravenous glucose-insulin-potassium (GIK). In acute myocardial infarction, GIK is undergoing a resurgence of interest due to the promising results of the large recent Argentinian trial. GIK acts in several ways, including the beneficial effects of insulin itself upon reperfusion, promotion of glycolysis, and inhibition of circulating fatty acids and hence of fatty acid oxidation. Metabolic therapy acting to protect the ischemic cell deserves more attention. PMID- 10665329 TI - [Congestive cardiomyopathies originating from arrhythmia]. AB - Arrhythmia-induced cardiomyopathy is partially or totally reversible left ventricular dysfunction after normalisation of the tachycardia or arrhythmia. On the one hand, there are pure forms in which the arrhythmia occurs in apparently normal hearts and, on the other hand, the more common form in which there is minimal underlying cardiac disease associated with the arrhythmia. Total or partial recovery after reduction of the arrhythmia or "ablation" of its substrate remains a key feature of the diagnosis. Many experimental studies of the functional and structural myocardial and neurohormonal effects of prolonged tachycardias or tachyarrhythmias have provided insight into the modes of occurrence and the characteristics of this type of "reversible" left ventricular dysfunction. But, in fact, there is a lack of anatomical, clinical and follow-up data of this syndrome, the diagnosis of which is always difficult and essentially retrospective after recovery of left ventricular function. PMID- 10665330 TI - [Compression of the renal artery by a musculo-tendinous band: an unrecognised cause of renovascular hypertension]. AB - Stenosis of a renal artery by extrinsic compression is an uncommon cause of renovascular hypertension. In rare cases, this compression is due to the presence of fibres from the diaphragm or the psoas muscle. This aetiology should be considered when renal artery stenosis is observed in a young hypertensive patient without cardiovascular risk factors. Spiral CT scan is particularly useful for studying the relationship between the diaphragm and arterial structures. Once the diagnosis has been made, the treatment is surgical section of the fibrous tissues responsible for the compression. PMID- 10665331 TI - [Troponins: new biochemical markers of myocardial damage. Structure, immunoassay and clinical application]. AB - The development of immunoassay techniques has made it possible to use new biochemical markers with a high myocardial specificity. Immunoassays have been developed for two of the structural myocardial proteins: troponin I and troponin T. The authors describe the biochemical properties of the troponins and the available methods of immunoassay. The main clinical applications of troponin measurement in cardiological practice are also reported. PMID- 10665332 TI - [Pre-capillary pulmonary hypertension complicating CREST syndrome. Apropos of a case]. AB - Pre-capillary pulmonary hypertension was the presenting sign of a CREST syndrome in a 65 year old woman. The diagnosis of this form of scleroderma is based on the association of a number of features (calcinosis, Raynaud's phenomenon, oesophageal dyskinesia, sclerodactylia and telangectasia). Scleroderma is the systemic disease which is usually complicated by pre-capillary pulmonary hypertension. This complication is observed in about 13% of CREST syndromes, but very rarely as severe pre-capillary pulmonary hypertension. The diagnosis of pre capillary pulmonary hypertension carries a poor prognosis with a 2 year survival rate of about 40%. Treatment is usually with calcium inhibitors but with no effect on prognosis. The use of prostacycline and its analogue, iloprost, is an interesting therapeutic strategy, currently under evaluation. Cardiopulmonary transplantation is the only treatment of very severe forms, despite the progressive character of the condition. All cases of pre-capillary pulmonary hypertension require complete aetiological investigation to exclude a systemic disease, especially a scleroderma and, above all, a CREST syndrome. PMID- 10665333 TI - [Immediate collateral coronary circulation after a methylergometrin test]. AB - The development of a collateral coronary circulation has been well studied by angiography in two main clinical situations: myocardial infarction (by durable coronary occlusion) and angina (due to significant coronary artery stenosis), but only rarely in spastic angina. The authors report the case of severe spasm at the site of non-significant stenosis after a methylergometrine test, with immediate contro-lateral collateral circulation in a patient with a short history of spastic angina without myocardial infarction. This observation demonstrates that collateral circulation may develop very rapidly in spastic angina (without basal ischaemia in the absence of significant coronary stenosis), because this patient only had seven ten-minute episodes of clinical ischaemia. As collateral circulation may mask clinical and electrical signs in spastic angina, this case suggests that angiographic control should be systematic during the methylergometrine test. PMID- 10665334 TI - [Pheochromocytoma simulating myocardial infarct. Apropos of a case]. AB - The authors report the case of a pheochromocytoma in a 67 year old man in whom the initial clinical presentation suggested myocardial infarction. Pheochromocytoma is usually an adrenal tumour with a very variable clinical symptomatology. It is very rare for cardiac disease to be a presenting symptom. The diagnosis was suggested by major blood pressure abnormalities occurring after starting medical treatment for infarction. Pheochromocytomas may cause serious cardiovascular disorders. The diagnosis must be suspected in the presence of atypical signs, an essential requirement to reduce the mortality of the disease. PMID- 10665335 TI - [Simultaneous utilization of an implantable automatic defibrillator in a patient with previously implanted bi-ventricular pacemaker for end-stage heart failure]. AB - We report a case of implantable cardioverter defibrillator (ICD) use after previous biventricular pacemaker insertion for end-stage heart failure. A 75-year old man with inoperable three-vessel coronary artery disease, permanent atrial fibrillation and end-stage heart failure underwent bi-ventricular pacemaker insertion and His bundle ablation for symptomatic control. NYHA class decreased from class III to II after this procedure. Four months after implant the patient developed paroxysmal sustained, symptomatic ventricular tachycardia. ICD implantation was undertaken. No potentially serious ICD-pacemaker interaction was noted during subsequent follow-up. We conclude that ICD implantation is feasible after previous bi-ventricular pacemaker insertion, without the need to explant the bi-ventricular pacing device. PMID- 10665336 TI - Human obesity and thinness, hyperlipidemia, hyperglycemia, and insulin resistance associated with HIV1 protease inhibitors. Prevention by alternating several antiproteases in short sequences. AB - In 1997, and mainly in 1998 and 1999, a lipodystrophic syndrome with central obesity, peripheral fat loss, hyperlipidemia, hyperglycemia and insulin-resistant diabetes II, was described as the most frequent manifestation of toxicity of HIV1 virostatic therapy, associated with protease inhibitors (PI) in 83% of the patients who used them for 10 months. Almost similar syndromes had been published before the latter, due, for example, to graft vs host reaction, or autoimmunity against insulin receptors, or to caloric excess in the presence of androgens (the mediator being hyperinsulinemia). Carr and Cooper have presented an original pathophysiological mechanism for the PI-associated syndrome, residing in 63% homology between HIV1-protease and the 3-low-density-lipoprotein-receptor-related protein (LRP), and in 53% homology between this virus enzyme and retinoid-binding protein type 1 (CRAB1). The treatment should be more subtle than those of common obesity and/or type II diabetes. This HIV1-protease inhibitor toxicity can be prevented by alternating several antiproteases in short sequences of the different ones. PMID- 10665337 TI - Prader-Willi syndrome, diabetes mellitus and hypogonadism. AB - Diabetes mellitus is not a diagnostic criterion for Prader-Willi syndrome (PWS), but it is often found in PWS patients. The etiology for diabetes mellitus in PWS may be related to the morbid obesity and consequent insulin resistance, because a decrease of oxytocin neurons and leptin resistance in PWS may cause hyperphagia, inducing obesity. However, treatment with growth hormone (GH) is beneficial for the majority of GH-deficient PWS children, because relative decreased fat mass and increased fat-free mass could prevent obesity and concomitant insulin resistance. Hypogonadism is thought to be due to hypogonadotrophic hypogonadism in a majority of PWS patients. Hypergonadotrophic hypogonadism secondary to cryptorchidism and its treatment is shown in other cases. Low luteinizing hormone and high follicle-stimulating hormone levels in PWS cases in young men with idiopathic oligospermia or in the early stages of puberty is less frequently reported. PMID- 10665338 TI - Plasma plasminogen activator inhibitor 1, insulin resistance and android obesity. AB - Plasma plasminogen activator inhibitor 1 (PAI-1) levels are elevated in insulin resistant subjects and are associated with increased cardiovascular risk of atherothrombosis. Strong association between PAI-1 and the metabolic components of the insulin resistance syndrome is found in clinical studies, suggesting that insulin resistance may regulate circulating PAI-1. However, the mechanisms underlying increased PAI-1 levels in such conditions are still poorly understood. Several studies have been carried out specifically in patients with central or android obesity, a major characteristic of the insulin resistance syndrome, and have suggested that visceral adipose tissue may be the major component of the relationship between android obesity and PAI-1. Accordingly, adipose tissue PAI-1 production was found to be elevated in obese human subjects, particularly in visceral adipose tissue. The genetic background for having high PAI-1 levels in several populations have been looked for and its role appeared to be weaker than that of the metabolic condition. High plasma PAI-1 levels are then clearly related to android obesity and insulin resistance, but the mechanisms whereby PAI 1 increases in plasma in these diseases remain to be determined. PMID- 10665339 TI - Lipoprotein (a) in android obesity and NIDDM: a new member in 'the metabolic syndrome'. AB - The 'metabolic syndrome' is a special clinical entity characterized by upper body segment obesity (android obesity), together with one or more of a constellation of metabolic disorders that includes glucose intolerance, which may amount to frank diabetes mellitus, hypertension, cardiovascular lesions, hyperuricemia, and dyslipidemias (hypercholesterolemia, hypertriglyceridemia and reduced serum HDL). Recently, lipoprotein (Lp) (a) proved to be a new member in this syndrome. Lp(a) has the distinctive feature of containing apolipoprotein (a), which is a glycoprotein linked to apo B100, and has a similarity to plasminogen; it is also structurally related to LDL. Lp(a) is a macromolecular complex which is genetically determined, and has been identified as an independent risk factor for premature coronary artery disease (CAD). It is elevated in diabetic and non diabetic android obese subjects, and aggravates the atherogenic effect of diabetes mellitus. Lp(a) is poorly influenced either by dietary measures or by hypolipidemic drugs. Unfortunately, few pharmacologic agents, such as niacin, nicotinic acid, sex hormones (estrogen and testosterone), alcohol and neomycin, affect Lp(a). PMID- 10665340 TI - Role of protein tyrosine phosphatase-1B in diabetes and obesity. AB - Type 2 or non-insulin-dependent diabetes mellitus (NIDDM) is reaching epidemic proportions in industrialized countries. Obesity is a major factor in this disease, since about 75% of obese individuals will develop type 2 diabetes. There is an urgent need to develop new therapies for these diseases. Recently, the protein tyrosine phosphatase PTP-1B has been shown to be a negative regulator of the insulin signaling pathway, suggesting that inhibitors of this enzyme may be beneficial in the treatment of type 2 diabetes. Mice lacking PTP-1B are resistant to both diabetes and obesity. PMID- 10665341 TI - In-vivo delivery of therapeutic proteins by genetically-modified cells: comparison of organoids and human serum albumin alginate-coated beads. AB - We have designed a self-assembling multimeric soluble CD4 molecule by inserting the C-terminal fragment of the alpha chain of human C4-binding protein (C4bp alpha) at the C-terminal end of human soluble CD4 genes. This CD4-C4bp alpha fusion protein (sMulti-CD4) and two other reference molecules, a fusion protein of human serum albumin (HSA) and the first two domains of CD4 (HSA-CD4) and monomeric soluble CD4 (sMono-CD4), were delivered in vivo by genetically modified 293 cells. These cells were implanted in mice as organoids and also encapsulated in HSA alginate-coated beads. sMulti-CD4 showed an apparent molecular weight of about 300-350 kDa, in accordance with a possible heptamer formula. sMulti-CD4 produced either in cell culture or in vivo in mice appeared to be a better invitro inhibitor of HIV infection than sMono-CD4. Plasma levels of sMulti-CD4, HSA-CD4, and sMono-CD4 reached approximately 2,300, 2,700, and 170 ng/mL, respectively, 13 weeks after in-vivo organoid implantation, which had formed tumours at that time. This suggests that the plasma half-life of sMulti-CD4 is much longer than that of sMono-CD4. The 293 xenogeneic cells encapsulated in HSA alginate-coated beads remained alive and kept secreting sMono-CD4 or HSA-CD4 continuously at significant levels for 18 weeks in nude mice, without tumour formation. When implanted in immunocompetent Balb/c mice, they were rejected two to three weeks after implantation. In contrast, encapsulated BL4 hybridoma cells remained alive and kept secreting BL4 anti-CD4 mAb for at least four weeks in Balb/c mice. These results suggest the clinical potential of the C4bp multimerizing system, which could improve both the biological activity and the poor in-vivo pharmacokinetic performance of a monomeric functional protein like soluble CD4. These data also show that a systemic delivery of therapeutic proteins, including immunoglobulins, can be obtained by the in-vivo implantation of engineered allogeneic cells encapsulated in HSA alginate-coated beads. PMID- 10665342 TI - Why have ten or so nontoxic, retrovirus integrase inhibitors not been made available for AIDS treatment? A ten-year experience [correction of experiment] must liberate them. AB - We detected in 1989, with the inhibitor test of proviral insertion into c-erb B erythroblastosis, two retrovirus integrase inhibitors: hydroxy-methyl-ellipticine and acriflavine. They have been used for ten years in AIDS patients with high efficacy and no toxicity. Since vitamin B12 and cobalt, which it contains, have been detected as HIV1-integrase inhibitors by an in-vitro test, we have also used vitamin B12 (combined with folic acid), whose clinical action has been remarkable. Ten or so other compounds have been detected by such in-vitro tests, among which there are many compounds (such as flavones) which are used in many conditions and are not toxic. PMID- 10665343 TI - Pediatric extracranial applications of MR angiography. AB - Extracranial magnetic resonance angiography (MRA) was performed in 20 children to evaluate for various arterial and venous conditions. Time-of-flight and phase contrast angiograms were constructed using a maximal-intensity-projection algorithm. The accuracy of MRA was comparable to Doppler ultrasound (n = 12) and conventional angiography (n = 3). MRA could provide an excellent mapping of patent (including collaterals) versus thrombosed vessels at sites not evaluated or inaccessible by sonography. Limitations included assessment of small and/or tortuous vessels, severely stenotic lesions, and very slow flow. PMID- 10665344 TI - Significance of resistive index in color Doppler ultrasonogram: differentiation between benign and malignant breast masses. AB - The objective of this article is to evaluate the significance of resistive index in differentiation between benign and malignant breast lesions on duplex ultrasonographic examination. Resistive indices obtained in 106 breast lesions of 104 patients were included. Sixty-four were benign (mean age: 32.4 +/- 11.1 years), and 42 were malignant lesions (mean age: 47.8 +/- 11.4 years). The resistive index was classified as follows: below 0.49, from 0.5 to 0.59, 0.6 to 0.69, 0.7 to 0.79, and above 0.8. We analyzed and defined the optimal threshold value of RI between benign and malignant lesions. The mean values of the RI of benign and malignant lesions were 0.62 +/- 0.095 (range 0.44-0.86) and 0.74 +/- 0.097 (range, 0.50-0.92), respectively. The resistive index exceeded 0.7 in 80% of malignant lesions. The difference of the RI between malignant and benign lesions was statistically significant when the threshold value was 0.7 (P < 0.001). A resistive index over 0.7 may suggest malignant lesions. Due to the considerable overlap of the range of the RI, it may not be diagnostic in any single patient; however, it may be helpful in conjunct with gray-scale image. PMID- 10665345 TI - Atypical presentation of dissection of the ascending aorta in young men with cystic medial necrosis: MR findings. AB - Dissection of the ascending aorta is usually associated with severe chest and/or back pain. We describe three young men, with pathologically proven cystic medial necrosis, who presented with atypical clinical symptoms and ascending aortic dissection diagnosed by MR imaging and surgery. Patients with cystic medial necrosis and aortic dissection may not present with a classic acute chest pain syndrome. PMID- 10665346 TI - Gastrocolic fistula due to adenocarcinoma of the colon: simulation of primary gastric leiomyosarcoma on CT. AB - This article describes the CT findings in two patients with adenocarcinoma of the colon and gastrocolic fistula which simulated the classic appearance of gastric leiomyosarcoma on CT. The role of CT in the diagnosis of gastrocolic fistula is also discussed. PMID- 10665347 TI - Rapidly and infiltratively growing Crohn's carcinoma of the small bowel: serial radiologic findings and a review of the literature. AB - We carried out a retrospective evaluation of serial changes in the small bowel radiographs of a patient with small bowel cancer accompanied by long-standing Crohn's disease. During the 8 months before diagnosis, marked morphological changes were noted. A solitary and irregular protrusion, and rapidly growing stricture under careful medical management of the underlying disease may indicate the development of cancer. PMID- 10665348 TI - Contrast-enhanced hepatic MRI: comparison of half-dose and standard-dose gadolinium DTPA administration in lesion characterization with T1-weighted gradient echo sequences. AB - The objective of this article was to compare half-dose (0.05 mm/kg) gadolinium enhanced dynamic hepatic MR imaging to standard doses (0.10 mm/kg). Eighteen patients for follow-up hepatic MR received 0.05 mm/kg of gadolinium DTPA dynamically with gradient-echo imaging. Imaging parameters were identical to a 0.10-mm/kg study; patients were imaged during multiple phases of contrast enhancement. Two readers assessed for enhancement patterns and characterization. Quantitative signal-to-noise ratios (S/N) were obtained for abdominal viscera and contrast-to-noise ratios (C/N) were obtained on up to three lesions. No significant difference for the arterial dominant phase (P > 0.05) was found. Significant differences were found in all categories during the portal venous phase (except pancreas) and equilibrium phase (except liver). Lesion C/N ratios were not significant at any point (P > 0.05). Sixty-two out of 64 lesions (97%) were identically characterized. Therefore, half-dose dynamic gadolinium-enhanced MR may have diagnostic value. PMID- 10665349 TI - Association of posterior rib fractures with exaggerated kyphosis and sternal collapse. AB - The ribs, sternum, and vertebrae all play an important role in stabilizing the thorax. Failure of one of these components places additional stress on the other supporting structures. We present a case of a 62-year-old man with multiple myeloma and osteopenia who sustained fractures to all three components. PMID- 10665350 TI - Avascular necrosis of bone in human immunodeficiency virus infected patients. AB - The objective of this article was to delineate the causes of avascular necrosis (AVN) in patients with human immunodeficiency virus (HIV). HIV-infected patients with pain in large joints were prospectively screened. Patients had radiographs and magnetic resonance imaging of their affected joints. Serum lipids, anticardiolipin antibody levels (IgG, IgM), and hemoglobin electrophoresis were performed on all patients who had radiographic studies. Medical records were screened for factors known to predispose for AVN. Eight patients completed the protocol, and five patients had AVN in seven joints. No common laboratory abnormality was identified in the patients with AVN. All of the patients with AVN had a history of steroid use; four of five patients having taken steroids for HIV related diseases. The cause of AVN does not appear to be directly related to the disease, but to steroid treatment for manifestations of the disease. PMID- 10665351 TI - Water-suppression MRI: role in the evaluation of osseous lesions. AB - The efficacy of chemical shift based water-suppression MRI in the evaluation of bone marrow lesions has not been previously reported. T1-weighted images without and with water suppression were compared in five patients with 16 lesions. There was a significant improvement in the contrast-to-noise ratio (from 4.32 to 5.95, P < 0.01) and contrast ratio (from 1.71 to 5.69, P < 0.004) with water suppression. Water suppression may be useful clinically by increasing the conspicuity of bone marrow lesions. PMID- 10665352 TI - Current status of burn resuscitation. AB - Rapid assessment and management of airway and breathing problems are required in the patient with severe burns complicated by significant facial burns and inhalation injury. A policy that results in intubation of all patients at potential risk for airway compromise can be both foolish and dangerous. At the same time, it is recognized that intubation of patients who are likely to develop unstable airways is necessary if transport times to burn centers are long and if i.v. resuscitation is initiated during transport. The ideal burn resuscitation formula does not exist. Whichever formula is used, patients must be monitored closely and the fluid resuscitation individualized according to their responses. Patients with delay in resuscitation, associated trauma, inhalation injury, or alcohol abuse may require fluid resuscitations greater than those predicted. The goal is to maintain urine outputs in the range of 0.5 to 1 mL/kg/hr for adults and 1 to 1.5 mL/kg/hr in children. In patients with fluid requirements greater than 150% of that predicted by formula, the addition of colloid at 12 hours can reduce total fluid requirements and burn edema. Early placement of pulmonary artery catheters can be useful in patients with known myocardial dysfunction, age greater than 65 years, severe inhalation injury, or fluid requirements greater than 150% of that predicted by formula. PMID- 10665353 TI - Current status of burn wound pathophysiology. AB - Healing is a continuum that can be unpredictable. Despite many advances and understanding of the multiple cellular processes and molecules involved in burn wound healing, physicians and patients have yet to reap the full benefit of this knowledge. The advances have occurred in a very short period, and with the exponential growth of molecular biology techniques and transgenic animal models, our understanding and treatment of burn wound healing could change exponentially over the next 10 years. The goal must be to continue to improve functional outcomes for burn survivors just as we have conquered critical care management for acutely injured burn patients. PMID- 10665354 TI - Primary excision of the burn wound. AB - Early excision of burn eschar and wound closure significantly improves survival following major burn injury. Immediate primary excision performed by burn experienced surgeons in dedicated burn care facilities can reduce further morbidity and mortality, length of hospital stay and medical costs. Burn care at the millennium is evolving rapidly into a subcategory of trauma surgery, with burn patients increasingly being viewed as victims of major trauma who benefit most from immediate and definitive surgical correction of their injuries. PMID- 10665355 TI - The acute and subacute management of the burned hand. AB - Management of the severely burned upper extremity remains a significant challenge to the most experienced clinician. An understanding of the underlying mechanism that uncorrected could culminate in a negative outcome is the key to formulation of a successful treatment plan. Initial proper splinting, avoidance of edema, the appropriate sequencing and integration of physical therapy, and judicious surgical intervention, all considered within the framework of the individual patient, are the components of the treatment plan that yields the most consistently good results. PMID- 10665356 TI - Reconstruction of the burned hand. AB - There is no "cookbook" for reconstructing the burned hand. Multiple issues can color the chances for a successful outcome. What is the endpoint of surgical effort? Is it when the patient tires, becomes discouraged, or ceases to return? These questions are not rhetorical. Whereas an appendectomy cures appendicitis, no single surgical procedure or series of procedures cures burns. Many patients spend their lives searching to be as they were preinjury. Although physicians as healers do not want to destroy hope, ethics command that we attempt to keep these patients focused on reality. Although there is always something that could be done, judgment dictates what should be done. The major goals are early independence and resumption of preburn lifestyle for the patient. A thoughtful surgical plan set up in conjunction with the burn team and with timed goals gives the patient the best chance for success. PMID- 10665358 TI - Acute and reconstructive management of the burned eyelid. AB - Introduced in 1875, the use of full-thickness skin graft for release of a lower eyelid ectropion secondary to burn contractures remains a mainstay of current burn treatment. The authors address issues such as acute care, the exposed cornea, scar management, and surgical management of eyelid deformities. PMID- 10665357 TI - Splints and scar management for acute and reconstructive burn care. AB - Because patients assume a position of comfort during acute burn management, affected joints or regions must be splinted in positions of function to avoid contractures. Particularly with the increasing trend towards ambulatory burn care, close monitoring by the burn team and patient education is required to achieve the best functional result. Despite adequate initial care, contractures sometimes occur, requiring a shift in splinting tactics to correct the deformity. If secondary reconstruction is required, the affected region is once again maintained in the position of function. Although the physiologic mechanism is incompletely defined, pressure therapy to prevent and treat hypertrophic burn scars is an integral component of burn care. Multiple materials and methods are available with treatment, starting soon after burn wound closure, and modified as needed until scar maturation has occurred. PMID- 10665359 TI - Reconstruction of the burned nose and ear. AB - Patients who have survived thermal injuries to the face suffer from severe disfigurement. When the nose and ear are involved, the resulting deformities are immediately obvious to all who see the patient. This level of injury results in a self-imposed confinement; the patients never leave their homes. It is therefore important that we plastic surgeons know, understand, and use all options available to improve our patients' appearance and ultimately their mental and physical well being. PMID- 10665360 TI - Reconstruction of the burned breast. AB - Fundamental principles of management of breast burns begin with recognition and preservation of any viable breast bud tissue. Reconstruction begins when the burned breast envelope is insufficient to allow unrestricted breast development. Complete contracture release is obtained by incision or excision of the restricting burn scar and thick split-thickness grafting. Occasionally, breast mound reconstruction with regional musculocutaneous flaps or tissue expanders is necessary. Balancing procedures, such as reduction or mastopexy of an opposite unburned breast, are often helpful. After a period of 6 to 12 months of compression garments, scar management, and settling, nipple-areola reconstruction is undertaken and consists of a combination of local flaps, full-thickness grafting, or composite grafts tailored to each patient's needs. Long-term follow up is necessary to ensure that breast development continues satisfactorily and that contractures do not recur. PMID- 10665361 TI - Tissue expansion in head and neck burn reconstruction. AB - The advent of tissue expansion has provided a useful tool for the reconstructive burn surgeon. As with many new techniques, there was an initial wave of enthusiasm surrounding the introduction of tissue expansion to burn reconstruction in the 1980s. High complication rates and many dissatisfying results followed. After early widespread use of tissue expansion, the authors have settled on a more refined approach to the reconstruction of head, neck, and facial burns. Today, head and neck burn reconstruction is accomplished best with a combination of skin grafting, local flaps, and occasional free flaps in addition to tissue expansion. In carefully selected head and neck burn patients and in many burn alopecia patients, tissue expansion can provide excellent functional and aesthetic results, with minimal donor site morbidity. PMID- 10665362 TI - Electrical burns. AB - Electrical burns can be divided into flash or typical thermal injury and high tension injury. The latter is usually caused by greater than 1000 volts and produces a clinically characteristic entry and exit wound. The optimal management of patients with high-tension electrical injury has evolved into a plan of urgent exploration and debridement, aggressive redebridement, and early wound closure. PMID- 10665363 TI - Reconstruction of the burned foot. AB - Burns of the feet pose unique and difficult problems in initial management, reconstruction, and the attainment of long-term functional results. The primary reconstructive goals for this region are unimpeded ambulation and weightbearing on a pain-free limb. These objectives can be achieved by adherence to established principles of wound management, a clear delineation of the reconstructive requirements of the foot, and a team approach toward attaining these goals. The goal of any method of reconstruction should be the restoration of function within a reasonable aesthetic appearance. With the foot, in addition to adequate healing, the goal should be the ability to walk again, wear normal footwear, and, if possible, return to work. PMID- 10665364 TI - A simple procedure for polymerase chain reaction of the PSBA gene in algae: application to the screening of mutations conferring atrazine resistance and discrimination of natural populations of Porphyra linearis. AB - A simplified procedure is described for polymerase chain reaction (PCR) of a partial sequence (bp 601-893) of the plastid gene psbA in the rhodophyte Porphyra linearis and the diatoms Haslea ostreria and Skeletonema costatum. This procedure involves the use of all tissues of P. linearis and live cell suspensions of H. ostreria or S. costatum, as DNA templates, without any further purification of DNA. As in the case of PCR with DNA extracts, a single major band of the expected size (292 bp) was obtained after PCR for the three species. Sequences of the amplified fragments were aligned, confirming that the amplified products were part of the psbA gene. The method was then used to screen mutations in partial psbA genes of 23 samples of P. linearis collected at four different stations along the mid-Atlantic coast of France. An alignment was obtained indicating the existence of mutations, though not in codons known for herbicide resistance. All mutations found were silent. However, genetic polymorphism discriminated between samples collected from two stations. The method employed allows rapid amplification of the herbicide target gene and simplifies the procedure for screening mutations or populations in algae. Its application to other genes and species is considered. PMID- 10665365 TI - Effect of turkey (Meleagridis gallopavo) breeder hen age and egg size on poult development. 1. Intestinal growth and glucose tolerance of the turkey poult. AB - Three experiments were conducted to determine if turkey (Meleagridis gallopavo) hen age and egg weight affect poult intestinal development and glucose tolerance during the first week after hatching. Differences in glucose tolerance were not consistent across the experiments. In experiment 1, 4-day-old poults from the younger hens and lightest egg weight class had significantly greater fasting plasma glucose concentrations (P < 0.04) and were also higher at 30 and 60 min post-injection of 250 mg glucose. In experiment 2, egg weight class had no significant effects on plasma glucose concentrations of 4-day-old poults (after injection of 2.5 mg glucose/g body weight). In experiment 3, plasma glucose concentrations were not different between groups at 0 or 30 min post-injection (3.75 mg glucose/g body weight), however, poults from the younger hens had 79-90 mg/dl higher plasma glucose concentrations 60 min post-injection versus poults from the older hens. Neither egg weight class or hen age consistently affected small intestinal weight, length, or density (g/cm) measures across experiments. PMID- 10665366 TI - Effect of turkey (Meleagridis gallopavo) breeder hen age and egg size on poult development. 2. Intestinal villus growth, enterocyte migration and proliferation of the turkey poult. AB - Villus growth, enterocyte migration and proliferation were measured in the small intestine of poults (Meleagridis gallopavo) to determine if hen age and/or egg size influences these characteristics during the first week after hatching. At hatching, distal jejunal villi were 22.8 microns longer in poults from the older (48 weeks) versus the younger (34 weeks) hens (P < 0.05). Similarly, labeled enterocytes in distal jejunal sections from poults from the older hens had migrated 28 microns (10%) farther along the crypt-villus axis at hatching, as compared to poults from the younger hens (P < 0.05). Villus growth differences and enterocyte migration were not consistently affected by hen age or egg weight class in poults from 1 to 7 days old. These results suggest that even though intestinal villi may be more advanced developmentally at hatch in poults from the older hens, however post-hatch growth of the intestine or the poult is not affected by hen age or egg weight class. PMID- 10665367 TI - Serum alpha- and gamma-tocopherols, retinol, retinyl palmitate, and carotenoid concentrations in captive and free-ranging bottlenose dolphins (Tursiops truncatus). AB - Concentrations of retinol, retinyl palmitate, beta-carotene, alpha-carotene, cryptoxanthin, lutein, lycopene, alpha-tocopherol, and gamma-tocopherol were measured in blood samples collected from 15 captive and 55 free-ranging bottlenose dolphins (Tursiops truncatus). From June 1991 to June 1994, blood samples were collected from captive animals residing at two locations; at Seven Seas (Brookfield Zoo, Brookfield, IL) and Hawk's Cay (Marathon Key, FL). Blood samples were collected from free-ranging animals from June 1991 to June 1996. Retinol levels were not significantly different between captive dolphin groups. However, Seven Seas animals had higher (P < 0.01) serum retinol concentrations compared to free-ranging animals (0.061 vs 0.041 microgram/ml). Retinyl palmitate was not detected in the serum of captive or free-ranging dolphins. Alpha tocopherol levels were significantly (P < 0.05) higher for Seven Seas dolphins (16.4 micrograms/ml) than for Hawk's Cay (13.0 micrograms/ml) and free-ranging dolphins (12.5 micrograms/ml). Gamma-tocopherol concentrations were similar among captive and free-ranging dolphins. Free-ranging dolphins showed levels of circulating carotenoids (lutein and beta-carotene) while the captive animals did not. Additional carotenoids (lycopene, alpha-carotene and cryptoxanthin) were analyzed but not detected in any samples. Serum vitamin differences between captive and free-ranging dolphins may reflect the natural diet or indicate some potential biological or nutritional status significance. PMID- 10665368 TI - Comparison of phosphodiesterase isozymes in rodent parotid glands. AB - We investigated phosphodiesterase (PDE) isozymes, which hydrolyze cAMP, in rodent parotid glands (mouse, hamster and guinea pig) in order to clarify the effects of cGMP and Ca/calmodulin on the regulation of cellular cAMP and compared them with those of the rat. More than 80% of the activities were in the supernatant fractions except for the hamster. The isozymes were fractionated using Mono Q ion exchange column. The mouse parotid PDEs consisted of PDE1 (Ca/calmodulin dependent), PDE2 (cGMP-stimulated), PDE3 (cGMP-inhibited) and PDE4 (cAMP specific) similar to those of the rat. PDE3 was not detected in the hamster, and PDE4 was not detected in the guinea pig. PDE activities in the supernatant of the mouse and the hamster were stimulated by cGMP, and that of the guinea pig was stimulated by Ca/calmodulin. These results suggest that various PDE isozymes are present in the parotid gland of several species of order Rodentia. There seems to be differences among the species with regard to the PDE isozymes. PMID- 10665369 TI - Influence of dietary protein on insulin-like growth factor binding proteins in the chicken. AB - We determined the effect of dietary protein on the distribution of insulin-like growth factor (IGF) binding proteins in chicken plasma. Three groups of male broilers (n = 6 per group) were fed (ad libitum) isocaloric diets containing 12, 21 or 30% dietary protein. Birds were fed respective diets beginning at 7 days of age and killed at 28 days. No differences were observed between adequate (21%) and high (30%) protein intakes for any of the parameters investigated (growth criteria, plasma levels of IGF-I, growth hormone or IGF-binding proteins). Feeding protein deficient diets (12%) resulted in a 34% decrease in body weight, 17% decrease in feed intake and a 39% increase in feed/gain ratio. IGF-binding proteins in plasma samples were separated by SDS-PAGE and transferred to nitrocellulose sheets. Nitrocellulose blots were probed with [125I]chicken IGF II. Four regions of binding activity corresponding to 70, 43, 30 and 24 kDa were observed in all samples. Birds consuming 12% dietary group protein had less than 50% of the 43-kDa binding activity of birds consuming 21 or 30% dietary protein. The 30-kDa binding activity was 42% lower in the 12% dietary protein group compared to birds consuming adequate protein. In contrast, 70- and 24-kDa binding activities were not influenced by dietary protein. Chickens consuming 12% dietary protein had higher levels of growth hormone and lower levels of IGF-I than those consuming 21 or 30% dietary protein. These data indicate that in chickens, the circulating levels of at least two independent IGF-binding proteins are influenced by dietary protein. PMID- 10665370 TI - The effect of urea exposure on isoaspartyl content and protein L-isoaspartate methyltransferase activity in Drosophila melanogaster. AB - Urea is a protein unfolding agent that can accumulate to locally high concentrations in tissues of many organisms. We used Drosophila melanogaster to test the hypothesis that urea loading would promote formation of isoaspartate (beta-carboxyl-linked aspartate), a common form of protein damage that occurs most readily in unstructured polypeptides and flexible regions of folded proteins. Ten populations of flies were tested; five control populations of urea sensitive flies and five previously selected urea-tolerant populations. We measured the effects of urea consumption on levels of both isoaspartate and protein L-isoaspartate methyltransferase (PIMT), an enzyme believed to function in the repair or removal of isoaspartyl proteins. For both sets of populations, urea feeding for 6 days increased isoaspartyl levels by approximately 60%, supporting the idea that disruption of protein secondary and tertiary structures can accelerate the formation of isoaspartate in vivo. Urea feeding tended to increase PIMT activity in both control and urea-tolerant populations. There were no significant differences in PIMT activities or isoaspartyl levels between the control and urea-tolerant flies raised on normal or urea food. The latter findings indicate that urea tolerance evolved in the selected populations without any significant change in PIMT expression or activity. PMID- 10665371 TI - Molecular cloning, sequence analysis and expression distribution of an aminopeptidase in Aplysia california. AB - We are investigating the role of membrane-bound peptidases in the inactivation of neuropeptides in Aplysia californica. Recently, we reported the biochemical characterization of a membrane-bound neuropeptide-degrading enzyme which has enzymatic characteristics similar to those of the mammalian aminopeptidase N (Bawab W, Querido E, Crine P, DesGroseillers L. Identification and characterization of aminopeptidases from Aplysia californica, Biochem J 1992;286:967-975). We now report the cloning and sequencing of a cDNA encoding an aminopeptidase enzyme (apAP) and the localization of the apAP transcript in Aplysia. The apAP cDNA encodes a putative protein of 1007 amino acids, which shows around 34% sequence identity to mammalian aminopeptidases A and N sequences. The deduced amino acid sequence suggests that apAP is a type II membrane-bound protein, with a long extracellular domain in which the consensus sequence of zinc-binding metallopeptidases (His-Glu-Xxx-Xxx-His) is found. RT-PCR and Northern blot experiments showed that the apAP gene is expressed as a single 6.8-kb transcript in the central nervous system, gill, heart, kidney and ovotestis. PMID- 10665372 TI - Comparative study on the fatty acid composition of two marine vertebrates: striped dolphins and loggerhead turtles. AB - The fatty acid composition of total lipids extracted from seven different tissues (fat, liver, cerebrum, cerebellum, lung, kidney and muscle) of 10 striped dolphins (Stenella coeruleoalba) (weight 75.7 +/- 16.2 kg) found dead or moribund during a morbillivirus epizootic, and from two tissues (fat and liver) of 54 loggerhead turtles (Caretta caretta) (18.5 +/- 10.5 kg) seized dead after illegal capture, both from the Mediterranean Sea, have been determined by high resolution chromatography techniques. When comparing both species, fatty acid composition is surprisingly similar, with a predominance of the monoenoic followed by the saturated group of fatty acids, and a very close ratio of polyunsaturates n-3/n 6. The relatively high arachidonic acid content in the liver of the two marine species is remarkable. The similar diet can play an important role in these findings, but it is suggested that probably metabolic pathways and essential fatty acid requirements between both marine vertebrates are similar, more than is expected from their earlier filogenetic evolutionary divergence. PMID- 10665373 TI - Thyroid hormones regulate lipid metabolism in a teleost Anabas testudineus (Bloch). AB - We compared the long-term action of 3,5,3'-triiodo-L-thyronine (T3) and 3,5 diiodo-L-thyronine (T2) on lipid metabolism in a teleost Anabas testudineus. Among the six groups of animals used in this experiment, except for the control group, all received 6-propylthiouracil (6-PTU) to create a hypothyroid state in order to analyse the action of iodothyronines on lipid metabolism. Injections of 6-PTU reduced T3 concentration in the circulation by 79.6% and injections of iodothyronines enhanced the level of T3 in the plasma, and a maximum increase was observed in T3 (500 ng)-treated specimens. Analysis of lipogenic enzymes in liver and heart showed that a tissue-specific variation exists in the action of thyroid hormones and, in many cases, activity is higher in T2-treated groups. Analysis of various lipid classes showed that long-term administration of T2 is also effective in producing a comparable effect with that of T3 on lipid metabolism. PMID- 10665374 TI - Modulation of reduction of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone by vitamin C-palmitate. AB - An in vitro study of effects of vitamin C-palmitate on the metabolism of 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in rat microsomes was performed. A sensitive assay method has been developed for the detection of metabolites of NNK in microsomes. Only the reduced metabolite of NNK, 4 (methylnitrosamino)-1-(3-pyridyl)-butanol (NNAL), was detected and measured in a time-course study. Vitamin C-palmitate enhanced the reduction of NNK in a concentration-dependent manner. The results indicate a significant increase in Vmax and K(m) in the presence of vitamin C. However, the rate of formation of NNAL at low substrate concentration varied. The ratio of Vmax to K(m) decreases. The results suggest that the kinetics are accounted for best by an uncompetitive activator binding model at low concentration of vitamin C. The uncompetitive binding model becomes sketchy at higher concentration of vitamin C. These observations infer that vitamin C loosely binds to the substrate-enzyme complex. Furthermore, the nature of the binding would facilitate the modulation of NNK biotransformation leading to the formation of NNAL. The results also show that vitamin C-palmitate is a potent activator of NNK reduction in rat liver microsomes. Thus, vitamin C-palmitate would mediate the metabolism of NNK through reduction. The resulting NNAL-glucuronide is more readily eliminated in urine. PMID- 10665375 TI - Purification and characterization of chitin-binding proteins from the hemolymph of sweet potato hornworm, Agrius convolvuli. AB - Three chitin-binding proteins (CBPs: CBP9, CBP15, CBP66) were identified from the larval hemolymph of sweet potato hornworm, Agrius convolvuli. Two (CBP9 and CBP15) of them have been isolated and purified by gel filtration (Superdex HR 75), cation-exchange chromatography (Mono S), and reverse-phase chromatography (muRPC PC 2.1/3). In experiments to detect CBPs in hemolymph, we examined whether ionic strength and existence of bovine serum albumin in the incubation solution influenced binding affinity of CBPs to chitin. The N-terminal sequences of three CBPs were determined by the automated Edman degradation and showed the sequence homology in basic local alignment search tool search CBP15 and CBP66 were quite similar to lysozymes and bovine serum albumins, respectively. In contrast, CBP9 is not similar to any other known protein, as judged from databank comparisons. Therefore, we concluded that CBP9 is a novel protein with binding capacity to chitin that is a component of the fungal cell wall. CBP9 has no antibacterial activity against Escherichia coli and Micrococcus luteus, and also showed negative response in hemagglutination assay. CBP9 is confirmed as a monomer with a molecular mass of 9.14 kDa by electron spray ionization and matrix-assisted laser desorption ionization mass spectrometry. PMID- 10665376 TI - D-aspartate oxidase is present in ovaries, eggs and embryos but not in testis of Xenopus laevis. AB - D-aspartate oxidase (DASPO) is an FAD dependent flavoprotein which catalyzes the oxidative deamination of D-aspartate using oxygen as electron acceptor. D aspartate and DASPO are supposed to be involved in the regulation of the central nervous system and in the animal development. This manuscript describes for the first time the presence of DASPO in Xenopus laevis fertilized eggs and embryos and suggests a different tissue distribution of this enzyme in adult male and female animals. In particular, by means of 2D-electrophoresis and affinity purified specific anti-DASPO antibodies, the enzyme was localized in fertilized eggs of X. laevis and in ovaries of adult animals but it was shown to be absent in the testis suggesting a gender specific expression. The protein from Xenopus ovaries has been purified by means of immunoprecipitation and it has M(r) of 30 kDa and pI of 8.1. PMID- 10665377 TI - Nitric oxide as a modulator of central respiratory rhythm in the isolated brainstem of the bullfrog (Rana catesbeiana). AB - Nitric oxide (NO) is a unique interneuronal neurotransmitter and/or neuromodulator that is involved in a variety of physiological functions within the central nervous system (CNS). In neural tissue, NO is generated from an oxygen-dependent, constitutive NO synthase (NOS) by glutamatergic stimulation of N-methyl-D-aspartate (NMDA) receptors. Recent studies indicate that NO has excitatory effects on breathing within the CNS and mediates a central component of the hypoxic ventilatory reflex in mammals. Because NMDA receptors are important in central respiratory rhythmogenesis, we hypothesized that NO would have significant effects on the central pattern generator (CPG) for breathing in the brainstem. To test this hypothesis, the effects of NO on respiratory-related neural activity were investigated using an in vitro brainstem preparation from North American bullfrogs (Rana catesbeiana). Extracellular recordings of respiratory-related burst activity were made from cranial nerves V, X and XII before and during superfusion of the brainstem with NO-generating compounds, or inhibitors of NO synthesis. Addition of the NO donor, sodium nitroprusside (SNP; 0.1-1.0 mM), or the amino acid precursor for NO synthesis, L-arginine (L-Arg; 0.01-1.0 mM), caused significant increases in respiratory-related burst frequency. Inhibition of NOS with N omega-nitro-L-arginine (L-NA; 5-10 mM), a non selective NOS inhibitor, caused a significant reduction in burst frequency or reversibly abolished neural activity. Brainstem perfusion with the specific neuronal NOS (nNOS) inhibitor, 7-nitro indazole (7-NI), produced significant, dose-dependent reversible reductions in burst frequency at concentrations of 0.1, 0.5 and 1.0 mM. These results suggest that production of NO, probably via nNOS, provides an excitatory input to the respiratory CPG in the amphibian brainstem. Our results suggest that NO may be a necessary inter- or intracellular messenger for neurotransmission and/or neuromodulation of central respiratory drive to motor effectors in the bullfrog. PMID- 10665378 TI - Central respiratory pattern generation in the bullfrog, Rana catesbeiana. AB - There are two components to breathing pattern generation the production of the pattern of neural discharge associated with individual breaths, and the pattern in which breaths are produced to effect ventilation. Bullfrogs typically breathe with randomly distributed breaths. When respiratory drive is elevated, breathing becomes more regular and often episodic. Studies on in vitro brainstem-spinal cord preparations of the adult bullfrog and in situ preparations of decerebrate, paralyzed, unidirectionally ventilated animals suggest that output from the central rhythm generator in frogs is conditional on receiving some input and that a host of central inputs remain even in the most reduced preparations. There appear to be descending inputs from sites in the dorsal brainstem just caudal to the optic chiasma that cluster breaths into episodes, a strong excitatory input caudal to this site but rostral to the origin of the Vth cranial nerve and, possibly, segmental rhythm generators throughout the medulla that are normally entrained to produce the normal breathing pattern. The data also suggest that the shape of the discharge pattern (augmenting, decrementing) and timing of outputs (alternating vs synchronous) associated with motor outflow during each breath are also dependent on the interconnections between these various sites. PMID- 10665379 TI - Learning, memory and a respiratory central pattern generator. AB - In an attempt to elucidate the causal mechanisms underlying learning and memory we have developed a model system, aerial respiration in the pond snail Lymnaea stagnalis. A three-neuron central pattern generator (CPG) whose sufficiency and necessity have been demonstrated mediates this behaviour. Aerial respiration, while an important homeostatic behaviour, is inhibited by the activation of the whole body withdrawal response that the animal uses to protect itself. We found that it was possible to operantly condition snails not to perform aerial respiration in a situation, a hypoxic environment, where aerial respiration should predominate. Operant conditioning was achieved by eliciting the pneumostome withdrawal response, part of the whole body withdrawal response, each time the animal attempted to open its pneumostome to breathe. Yoked control animals did not demonstrate an alteration in breathing behaviour. Subsequently we determined neural correlates of this associative behaviour and found that neuronal changes are distributed throughout the CPG. This preparation may afford us the opportunity to determine the casual neuronal changes that underlie learning and memory of associative conditioning. PMID- 10665380 TI - The fictively breathing tadpole brainstem preparation as a model for the development of respiratory pattern generation and central chemoreception. AB - Spontaneous high-frequency, low-amplitude and low-frequency, high-amplitude efferent bursting patterns of cranial and spinal motor nerve activity in the in vitro brainstem preparation of the bullfrog tadpole Rana catesbeiana have been characterized as fictive gill and lung ventilation, respectively (Gdovin MJ, Torgerson CS, Remmers JE). Characterization of gill and lung ventilatory activity in cranial nerves in the spontaneously breathing tadpole Rana catesbeiana, FASEB J 1996;10(3):A642; Gdovin MJ, Torgerson CS, Remmers JE. Neurorespiratory pattern of gill and lung ventilation in the decerebrate spontaneously breathing tadpole, Respir Physiol 1998;113:135 146; Pack AI, Galante RJ, Walker RE, Kubin LK, Fishman AP. Comparative approach to neural control of respiration, In: Speck DF, Dekin MS, Revelette WR, Frazier DT, editors. Respiratory Control Central and Peripheral Mechanisms. Lexington: University of Kentucky Press, 1993:52-57). In addition, the ontogenetic dependence of central respiratory chemoreceptor stimulation on fictive gill and lung ventilation has been previously described (Torgerson CS, Gdovin MJ, Remmers JE. Fictive gill and lung ventilation in the pre- and post-metamorphic tadpole brainstem, J Neurophysiol 1998, in press). To investigate the neural substrates responsible for central respiratory rhythm generation of gill and lung ventilation in the developing tadpole, we recorded efferent activities of cranial nerve (CN) V, VII, and X and spinal nerve (SN) II during changes in superfusate PCO2 before and after multiple transection of the in vitro brainstem. The brainstem was transected between CN VIII and IX and the response to changes in PCO2 was recorded. A second transection was then made between the caudal margin of CN X and rostral to SN II. Preliminary data reveal that robust gill ventilation was recorded consistently only if the segment of brainstem included CN X, whereas the loci capable of eliciting fictive lung bursting patterns appeared to differ depending on developmental stage. These data demonstrate that the neural substrate required for fictive gill and lung ventilation exists in anatomically separate regions such that the gill central pattern generator (CPG) is located in the caudal medulla at the level of CN X throughout development, whereas the location of the lung CPG is located more rostrally at the level of CN VII in the post-metamorphic larva. Both in vivo and in vitro studies revealed two distinct neural bursting patterns associated with gill and lung ventilation. Sequential activation of CN V, VII, X were observed during gill ventilation of in vivo and fictive gill ventilation in vitro, whereas these nerve activities, along with SN II displayed more synchronous bursting patterns of activation during lung ventilation and fictive lung breaths. PMID- 10665381 TI - Characteristic transport of lactoferrin from the intestinal lumen into the bile via the blood in piglets. AB - Lactoferrin is a major iron-binding protein in milk from several species, such as humans, monkeys, mice and sows. Using neonatal and weaner piglets, the characteristic transfer of lactoferrin from intestinal lumen into bile via the circulation was investigated. Bovine lactoferrin (1 or 3 g/kg body weight) was infused into the stomach through a polyethylene tube or into the duodenum through a duodenal catheter over 5 min. Peripheral blood and bile samples were collected after the infusion. Lactoferrin absorbed into plasma and bile were assayed quantitatively by double-antibody enzyme-linked immunosorbent assay, and homogeneity of bovine lactoferrin in plasma and bile was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting methods. Morphological investigation was carried out according to the peroxidase anti peroxidase method. Following oral administration in neonatal pigs, bovine lactoferrin appeared in the blood circulation and reached a peak level after 2 h. It was confirmed immunohistochemically that lactoferrin was transported by endocytosis via the epithelial cells. Lactoferrin absorbed into the blood was also detected in the bile and reached a peak value 12 h after oral administration. Transportation of lactoferrin from the intestinal lumen into the bile via the bloodstream was also observed in weaner piglets. Lactoferrin transported into plasma and bile was confirmed to be the same substance as administrated lactoferrin by electrophoresis and immunoblotting methods. Lactoferrin transported into bile was re-absorbed into the blood in neonatal pigs. These results demonstrate that lactoferrin contained in milk is transported into the circulation from the intestinal lumen and excreted into the bile, suggesting the possibility of entero-hepatic circulation of lactoferrin in neonatal pigs. PMID- 10665382 TI - Modulation of stress hormones in rainbow trout by means of anesthesia, sensory deprivation and receptor blockade. AB - Sympathetic activation leading to increased levels of blood catecholamines, and stimulation of the hypothalamic-pituitary inter-renal axis leading to increased cortisol, are difficult to avoid when handling animals. Yet, in research on effects of acute stress, elicitation of such responses must be minimized in the control groups. The work examines means to achieve a minimally disturbed state in rainbow trout (Oncorhynchus mykiss). Level of arousal was determined by adrenaline and cortisol concentrations in plasma, and by the spleen:somatic index. Fish were prepared for bleeding by rapid capture and concussion, by infusion of anesthetic into the undisturbed home tank, by confinement in black boxes, or by being fed alpha- and beta-receptor antagonists. Even when done quickly, netting and concussion yielded fish with ca. 200-pmol adrenaline/ml plasma. Cortisol was elevated (to > 10 ng/ml) within 30 s of stress initiation. Surreptitious infusion of anesthetic (2-phenoxyethanol, PE) into tanks yielded fish with lower adrenaline levels (means 19.34 and 19.58 pmols/ml in home tank and black boxes, respectively). Among fish given phentolamine and propranolol, spleen:somatic indices and plasma adrenaline were higher than in diet controls, whether undisturbed or stressed, indicative of successful receptor blockade. Since careful infusion of 2-PE yielded the lowest adrenaline levels, and requires no special apparatus, it is the method of choice for obtaining minimally stressed fish. PMID- 10665383 TI - Effects of thermal acclimation on nervous conduction and muscle contraction in the frog Rana temporaria. AB - The effects of season and acclimation temperature on the latency of the leg withdrawal reflex and three of its components have been studied: conduction velocity in the sciatic nerve, spinal conduction time, and contraction time of gastrocnemius muscle. The latency of the leg withdrawal reflex was markedly shortened by cold acclimation: the reaction times were at 6 degrees C 1.54 s in 4 degrees C acclimated and 3.97 s in 24 degrees C acclimated winter frogs. Also, the temperature dependence of the reflex latency was reduced by cold acclimation. Thus, frogs acclimated to cold responded to external stimuli in cold more rapidly than warm-acclimated ones. This cold adaptation of the reflex could not be explained by changes in its studied components. These made up only one-tenth of the reflex response time, and either did not show significant cold acclimation (muscle contraction and spinal conduction times in summer) or showed inverse acclimation, especially when measured at high temperatures (i.e. conduction velocities were reduced by acclimation to cold). Thus, the cold acclimation of the reflex response probably resides in the sensory component of the response. The inverse temperature adaptation response of conduction velocities may reflect a reduced ion permeability across cellular membranes in cold which decreases metabolic energy expenditure during inactive periods. PMID- 10665384 TI - Pre-existing bacterial infections, not stress fever, influenced previous studies which labeled Gerrhosaurus major an afebrile lizard species. AB - Early studies indicated that the Sudan plated lizard, Gerrhosaurus major, did not exhibit a febrile response when challenged with bacterial pyrogen. More recent results indicated that a 14-day antibiotic treatment regime produced a significant decrease (0.5 +/- 0.1 degree C) in the mean selected body temperature (MSBT) for this species (31.3 +/- 0.2-30.8 +/- 0.2 degrees C). The antibiotic treatment results suggested that G. major already had a fever caused by a pre existing bacterial infection. The current study was designed first to determine if a sub-population of G. major, with a higher mean pre-antibiotic treatment MSBT would exhibit a greater decrease in MSBT after antibiotic treatment. A 14-day antibiotic treatment regime for G. major (N = 7) with MSBTs > or = 31.9 degrees C (mean 32.4 +/- 0.2 degrees C) produced a significant decrease of 1.7 +/- 0.4 degrees C in MSBT to 30.7 +/- 0.3 degrees C. Analysis of the combined antibiotic treatment MSBT data from [13] and the current study demonstrated that the magnitude of the change in MSBT after antibiotic treatment was dependent upon the pre-antibiotic treatment MSBT. These data imply that animals with a greater pre treatment MSBT and greater MSBT change had a greater magnitude fever. In the second portion of this study the MSBT for individual lizards was measured during separate experiments using both indwelling cloacal thermocouples taped to the tail of the lizard and telemeters implanted into the peritoneal cavity of the lizard. This second study was designed to determine if measurement of Tb using thermocouples induced a stress fever which may have masked a portion of the pyrogen-induced fever. The MSBT measured using indwelling cloacal thermocouples (30.5 +/- 0.3 degrees C) was not significantly greater (T > 0.05) than the MSBT increased using telemeters (31.0 +/- 0.2 degrees C). The results of the experiments from this study demonstrate that the previously reported afebrile state for G. major was the result of animals having pre-existing bacterial infections. G. major does exhibit a febrile response similar to other lizard species. PMID- 10665385 TI - Respiratory consequences of feeding in the snake Python molorus. AB - Snakes can ingest large meals and exhibit marked increases in metabolic rate during digestion. Because postprandial oxygen consumption in some snakes may surpass that attained during exercise, studies of digestion offers an alternative avenue to understand the cardio-respiratory responses to elevated metabolic rate in reptiles. The effects of feeding on metabolic rate, arterial oxygen levels, and arterial acid-base status in the snake Python molorus are described. Four snakes (180-250 g) were cannulated in the dorsal aorta and blood samples were obtained during 72 h following ingestion of a meal (rat pups) exceeding 20% of body weight. Oxygen consumption increased from a fasting value of 1.71 +/- 0.08 to 5.54 +/- 0.42 ml kg-1 min-1 at 48 h following feeding, and the respiratory gas exchange ratio increased from 0.67 +/- 0.02 to a maximum of 0.92 +/- 0.03 at 32 h. Plasma lactate was always less than 0.5 mM, so the postprandial increase in metabolic rate was met by aerobic respiration. In fasting animals, arterial PO2 was 66 +/- 4 mmHg and haemoglobin-O2 saturation was 92 +/- 3%; similar values were recorded during digestion, but haematocrit decreased from 15.8 +/- 1.0 to 9.8 +/- 0.8 due to repeated blood sampling. Plasma [HCO3-] increased from a fasting level of 19.3 +/- 0.8 to 25.8 +/- 1.0 mmol l-1 at 24 h after feeding. However, because arterial PCO2 increased from 21.1 +/- 0.5 to 27.9 +/- 1.4 mmHg, there was no significant change in arterial pH from the fasting value of 7.52 +/- 0.01. Acid-base status returned to pre-feeding levels at 72 h following feeding. The increased arterial PCO2 is most likely explained by a reduction in ventilation relative to metabolism, but we predict that lung PO2 does not decrease below 115 mmHg. Although ingestion of large meals is associated with large metabolic changes in pythons, the attendant changes in blood gases are relatively small. In particular, the small changes in plasma [HCO3-] and stable pH show that pythons respond very differently to digestion than alligators where very large alkaline tides have been observed. It is unclear why pythons and alligators differ in the magnitude of their responses, but given these interspecific differences it seems worthwhile to describe arterial blood gases during digestion in other species of ectothermic vertebrates. PMID- 10665386 TI - Fate and distribution of 14C-atrazine in a tropical oxisol. AB - The herbicide atrazine is the most commonly detected pesticide in groundwater world-wide. A new microcosm test-system was used to determine the fate of 14C atrazine in a Brazilian oxisol. 14C Ring-labelled atrazine was applied in a mixture with the commercial product Gesaprim 500 (Novartis) at a rate of 3 kg ha 1. During two months, about 1% of the initially applied amount was lost by volatilization. The mineralization of the pesticide, measured directly using 14CO2 evolved from the applied pesticide, was between 0.09% and 0.16%, whereas less than 0.2% was leached. The distribution of radioactivity in the soil profile showed that most of the radioactivity remained in the top soil down to a 3 cm depth. The radioactivity in the upper 3 cm of the column was adsorbed perferably in fulvic acid (FA) and human fractions. PMID- 10665387 TI - The n-octanol and n-hexane/water partition coefficient of environmentally relevant chemicals predicted from the mobile order and disorder (MOD) thermodynamics. AB - The quantitative thermodynamic development of the mobile order and disorder theory in H-bonded liquids is extended in order to predict the partition coefficient. With respect to the classical predictive methods, the great advantage of the present approach resides in the possibility of predicting partition coefficient not only in the reference n-octanol/water partitioning system, but also in any mutually saturated two-phase system made up of two largely immiscible solvents. Constructed from the various free energy contributions encoded in the distribution process, the model furthermore provides a useful tool to understand both the origin and the factors, like the solute molar volume, that determine the partitioning of non-electrolytes between two immiscible liquid phases. From the comparison of the relative magnitude of the terms which contribute to the overall log P value, much information can also be gained concerning the variation of the partition coefficients of the same substances in different distribution systems. For example, the model has successfully been applied to the log P prediction of a number of environmentally important chemicals of varying structure, size and chemical nature in the n octanol/water and n-hexane/water systems. Whatever the complexing or non complexing substances studied, the hydrophobic effect always represent the driving force that rules distribution processes in the aqueous environments. As the dominant contribution to the partition coefficient in any organic/aqueous binary system, it is evidenced why hydrophobicity is usually considered to be a good measure of lipophilicity. PMID- 10665388 TI - Fuzzy clustering analysis of the first 10 MEIC chemicals. AB - In this paper, we discuss the classification results of the toxicological responses of 32 in vivo and in vitro test systems to the first 10 MEIC chemicals. In this order we have used different fuzzy clustering algorithms, namely hierarchical fuzzy clustering, hierarchical and horizontal fuzzy characteristics clustering and a new clustering technique, namely fuzzy hierarchical cross classification. The characteristics clustering technique produces fuzzy partitions of the characteristics (chemicals) involved and thus it is a useful tool for studying the (dis)similarities between different chemicals and for essential chemicals selection. The cross-classification algorithm produces not only a fuzzy partition of the test systems analyzed, but also a fuzzy partition of the considered 10 MEIC (multicentre evaluation of in vitro cytotoxicity) chemicals. In this way it is possible to identify which chemicals are responsible for the similarities or differences observed between different groups of test systems. In another way, there is a specific sensitivity of a chemical for one or more toxicological tests. PMID- 10665389 TI - Bisphenol A concentrations in receiving waters near US manufacturing and processing facilities. AB - Bisphenol A (BPA) (CAS 80-05-7) was analyzed in receiving waters upstream and downstream of US manufacturers (1996 and 1997) and processors (1997) during seasonal low flow periods. BPA was not detected (< 1 microgram/l) in any surface water sample in 1996 or at six of seven sites in 1997. Concentrations near the seventh site ranged from 2 to 8 micrograms/l; however, its receiving stream had no measurable flow and concentrations represent undiluted effluent. All surface water concentrations from this and other studies were less than the freshwater predicted no effect concentration (PNEC) of 64 micrograms/l, suggesting that BPA discharges from manufacturing and processing facilities to surface water do not pose an environmental concern. PMID- 10665390 TI - Volatile organic compounds in the surface waters of northern Greece. AB - An investigation into the occurrence of volatile organic compounds (VOCs) was conducted for a period of two years in the surface waters of Northern Greece. Samples from four rivers and five lakes were taken seasonally and analyzed for VOCs. The analysis has been performed by purge-and-trap (PAT) gas chromatographic mass spectrometric (GC-MS) technique. The most commonly encountered VOCs in surface waters were chloroform, carbon tetrachloride, trichloroethylene, dichlorobromomethane, tetrachloroethylene, and chlorodibromomethane. PMID- 10665391 TI - Nitro-PAH in ambient particulate matter in the atmosphere of Athens. AB - Nitrated polynuclear aromatic hydrocarbons (NPAH) with a molecular mass of 247 Daltons were found in soot collected in downtown Athens during a campaign performed in 1996. In particular, 2-nitrofluoranthene (2-NFa) and 2-nitropyrene (2-NPy), which are mainly related to photo-induced chemical processes occurring in the atmosphere, were more abundant than 1-nitropyrene (1-NPy) usually associated to motor vehicle exhaust. PMID- 10665392 TI - Comparison of an enzyme-linked immunosorbent assay (ELISA) to gas chromatography (GC)--measurement of polychlorinated biphenyls (PCBs) in selected US fish extracts. AB - The analysis of PCBs in fish tissues by immunoassay methods was evaluated using fish collected from a US monitoring program, the National Contaminant Biomonitoring Program of the US Department of Interior, Fish and Wildlife Service. Selected composite whole fish samples, which represented widely varying concentrations and sources of PCBs, were extracted and subjected to congener PCB analysis by gas chromatography (GC) and total PCB analysis using an ELISA (ePCBs) calibrated against technical Aroclor 1248. PCB congener patterns in these fishes were different from the patterns found in commercial Aroclors or their combinations as demonstrated by principal component analysis of normalized GC congener data. The sum of the PCB congeners measured by GC (total-PCBs) ranged from 37 to 4600 ng/g (wet weight). Concentrations of PCBs as determined by the ELISA method were positively correlated with total-PCBs and the ePCBs/total-PCBs ratios for individual samples ranged from 1 to 6. Ratios of ePCBs/total-PCBs for dilutions of Aroclors 1242, 1254, and 1260 and for matrix spikes range from 0.6 for 1242 to 2.5 for 1254 and 1260. These results suggest that higher chlorinated PCB congeners have higher affinity for the anti-PCB antibodies. Partial least squares with latent variable analysis of GC and ELISA data of selected Aroclors and fish samples also support the conclusion that ELISA derived PCB concentrations are dependent on the degree of chlorination. PMID- 10665393 TI - Biological denitrification in a closed seawater system. AB - Build-up of high nitrate concentrations in closed seawater systems where primary productivity is undesirable and water changes are impractical presents unique problems. Nitrate concentration in Ocean Tank at the New Jersey State Aquarium reached 9500 microM after 6 years of operation. A biological denitrification system was installed in 1998 and nitrate concentration in the aquarium decreased to 7000 microM within the first 100 days of operation. The system offers additional benefits by increasing the pH and alkalinity of seawater and providing a reducing environment to balance the oxidizing disinfection environment in the aquarium. The initial performance of the denitrification system was monitored and two semi-empirical models were developed: one based on the actual methanol additions, and another based on the daily amounts of nitrogen gas removed. The first model predicts a net nitrate decrease of 39 microM/day in the aquarium. The second model predicts a net decrease of 25 microM/day, in good agreement with the empirical value of 23 microM/day. This indicates that nitrogen gas removal is the controlling factor during denitrification in this facility, and the second model can be used to predict and optimize the operation of the system. PMID- 10665394 TI - Biodegradation of pyrene by sediment fungi. AB - Micromycetes were isolated from PAHS-contaminated sediment and identified. They were investigated for pyrene degradation (10 mg l-1) in liquid synthetic medium for two days. Among the 41 strains isolated, 10 highly degraded pyrene (> 2.4 mg g-1 dry weight): two Zygomycetes (Mucor racemosus, M. racemosus var. sphaerosporus), 6 Deuteromycetes (Gliocladium virens, Penicillium simplicissimum, P. janthinellum, Phialophora alba, P. hoffmannii, Trichoderma harzianum), a Dematiaceae (Scopulariopsis brumptii) and a Sphaeropsidale (Coniothyrium fuckelii). Zygomycetes appeared as one of the most efficient taxonomic groups, especially with Mucor racemosus. Penicillium crustosum was the only strain that did not degrade pyrene. Among the 10 fungi which were performant for pyrene degradation, nine were not yet reported in the literature and showed a real value for PAH remediation. PMID- 10665395 TI - Nitro musks, nitro musk amino metabolites and polycyclic musks in sewage sludges. Quantitative determination by HRGC-ion-trap-MS/MS and mass spectral characterization of the amino metabolites. AB - Nitro- and polycyclic musks were quantified in sewage sludge samples from different catchment areas using high-resolution gas chromatography (HRGC) and ion trap MS/MS. Collision induced dissociation (CID) turned out to be a useful tool for quantification of the analytes. Negative chemical ionization (NCI) quadrupole MS in the selected ion mode (SIM) showed similar sensitivities compared to ion trap MS/MS. Among the nitro musks, musk ketone (MK) and musk xylene (MX) were the main compounds in predominantly domestic sewage sludges, found at low microgram/kg dry matter (d.m.) whereas polycyclic musks were present in domestic as well as in industrial sludges up to 12 mg/kg d.m. Galaxolide (HHCB) and Tonalide (AHTN) were the major polycyclic musks found in the sludges. Amino metabolites of the nitro musks, amino musk xylene (AMA), amino musk moskene (AMM) and amino musk ketone (AMK) were detected for the first time in sewage sludges, and reached partly higher concentrations compared to the parent compounds. PMID- 10665396 TI - Degradation of 2,4,6-trinitrotoluene in water and soil slurry utilizing a calcium peroxide compound. AB - The degradation of 2,4,6-trinitrotoluene was examined in pure water and contaminated soil slurry using calcium peroxide as a source of solid hydrogen peroxide and oxygen. The extent of TNT oxidation was compared with that obtained by using hydrated lime, which is normally generated by slurrying CaO2 in water and contained in CaO2 technical formulation (approximately 50%, w/w). Complete TNT degradation occurred between 280 min, 0.1% CaO2/Ca(OH)2 and 20 min, 1% CaO2/Ca(OH)2. A large part of the generated oxidation products, 80-90%, were absorbed on the solid calcium hydroxide, whereas the remaining 10-20% was detected in solution until 48 h. Removal of nitro groups was extremely effective in CaO2 slurry, where all the nitrogen (3 mol per mol of TNT) was removed from TNT within 240 min. Respect to calcium hydroxide, the peroxy compound liberated H2O2 in solution, 370 mg l-1 at 0.2% CaO2, w/v, which then decomposed within 480 min. Most of the 14C-TNT was retained more strongly on the calcium hydroxide generated by slurrying CaO2. This pool remained adsorbed on the solid until pH dropped below 5.8. The treatment of a contaminated soil slurry, 700 mg TNT kg-1, reduced CH3CN extractable TNT below 20 mg kg-1 at very low concentration of CaO2/Ca(OH)2, approximately 0.2%, w/w. Both oxidants do not lead to soil sterilization as the phosphorus added to neutralize the pH serves as a source of nutrient for the soil biomass. PMID- 10665397 TI - Monitoring of bioremediation by soil biological activities. AB - An evaluation of soil biological activities as a monitoring instrument for the decontamination process of a mineral-oil-contaminated soil was made using measurements of microbial counts, soil respiration, soil biomass and several enzyme activities. The correlations between these parameters and with the levels of hydrocarbon residues were investigated; the effects of different N- and P sources on hydrocarbon decontamination and soil biological activities were determined. Inorganic nutrients stimulated hydrocarbon biodegradation but not all biological activities to a significant extent. Biodegradation could be monitored well by soil biological parameters: the residual hydrocarbon content correlated positively with soil respiration, biomass-C (substrate-induced respiration), and with activities of soil dehydrogenase, urease and catalase. Soil lipase activity and the number of hydrocarbon utilizers correlated negatively (P < 0.0001) with the remaining hydrocarbon content. PMID- 10665398 TI - Minimization of cobalt nuclide emissions in supercritical water oxidation of spent resin. AB - A rapid and complete destruction of organics in the supercritical water oxidation (SCWO) of the Co-exchanged resin was found experimentally. Due to an extremely low solubility of CoSO4 salt formed and separated effectively in the SCWO process, a minimal release of the nuclide Co would be warranted. In addition, recycling of Co nuclides is also possible by decomposition of the CoSO4 species at elevated temperatures (> 1040 K). PMID- 10665399 TI - Non-enzymatic reduction of azo dyes by NADH. AB - Nicotinamide adenine dinucleotide (NADH) reduces a variety of azo dyes by four electrons to generate the corresponding aromatic amines. This reduction is pH dependent and increases with decreasing pH. Reduction of 4-(4'-sulfophenylazo) phenol and 2-(4'-sulfophenylazo)-phenol, specifically substituted with methyl, methoxy, halo, and nitro groups, was examined to determine the susceptibility of azo dyes to reduction by NADH. Except for the nitro-substituted azo dyes, all other azo dyes were reduced. Possible mechanisms of reduction are proposed. The implications of our findings to microbial degradation and mammalian metabolism of azo dyes are discussed. PMID- 10665400 TI - Effects of heterocyclic PAHs (N, S, O) on the biodegradation of typical tar oil PAHs in a soil/compost mixture. AB - The interaction phenomena during the biodegradation of typical coal tar polycyclic aromatic hydrocarbons and their heterocyclic analogues (N, S, O) were investigated in an artificially contaminated AhA1-horizon/compost mixture. All compounds were partly or completely biodegraded. Degradation of two- to five-ring PAHs was inhibited by the presence of hetero-PAHs, whereas degradation of just some hetero-PAHs was inhibited by the presence of PAHs. Among the hetero-PAHs the sulphur-containing compounds were less susceptible to degradation than the corresponding oxygen- or nitrogen-containing analogues. The basic azaarene acridine showed an extreme persistence and strong sorption to the soil matrix proved by an increase of recovery after saponification of the soil matrix. PMID- 10665401 TI - Degradation and toxicity reduction of textile effluent by combined photocatalytic and ozonation processes. AB - To minimize the environmental impact of textile effluents, mainly related to their high coloration and the presence of toxic or carcinogenic reactive dyes, the efficiency of photochemical and ozonation processes, applied in the form of isolated and combined procedures, were evaluated. The investigation was focused on the reduction of total organic carbon content (TOC), color and acute toxicity (monitoring by inhibition of Escherichia coli respiration). For a reaction time of 60 min, the anatase TiO2-assisted photocatalytic process produces color and TOC reduction of about 90% and 50%, respectively. Meanwhile, the ozonation process gives a decolorization of about 60% but negligible TOC reduction. When the processes were applied in a simultaneous form, the decolorization was almost complete and the TOC reduction was higher than 60%. The three treatments studied yield an acute toxicity reduction of around 50%. PMID- 10665402 TI - The behaviour of N-(phenylsulfonyl)-glycine and phenacetin in a municipal sewage treatment plant--a case study. AB - The behaviour of N-(phenylsulfonyl)-glycine (PSG) and phenacetin (PHE) in a municipal sewage treatment plant near Heidelberg, Germany, was investigated in the summer of 1997. For that purpose, 10 g of each substance was dissolved and poured simultaneously into the influent. In addition to the spiked compounds, the samples of the influent, the biological stage and the effluent were analyzed for N-(phenylsulfonyl)-sarcosine (PSS), N-methyl-N-(phenylsulfonyl)-amide (MPS), N methyl-phenacetin, N-methyl-N-(phenylsulfonyl)-epsilon-aminocaproic (PSC) acid and its degradation product N-methyl-N-(phenylsulfonyl)-gamma-aminobutyric (PSB) acid. Within 24 h PHE could be detected almost quantitatively in the effluent. Since N-methyl-phenacetin could not be found in any of the samples, apparently no methylation of the amino-group of PHE took place. The amount of PSG in the effluent was within 24 h 26.0 g, which is more than two fold higher than added. The decrease of PSG between biological stage and effluent and the increase of PSS within the same time correlate well. Therefore, the formation of PSS by microbial methylation of PSG in the sewage treatment plant must be assumed. PMID- 10665403 TI - Fate of bromine in pyrolysis of printed circuit board wastes. AB - Behavior of Br in pyrolysis of the printed circuit board waste with valuable copper and oil recycling has been studied in the present work. Experimentally, pyrolysis of the printed circuit board waste generated approximately 40.6% of oils, 24.9% of noncondensible gases and 34.5% of solid residues that enriched in copper (90-95%). The cuts of the oils produced from pyrolysis of the printed circuit board waste into weighted boiling fraction were primarily light naphtha and heavy gas oil. Approximately 72.3% of total Br in the printed circuit board waste were found in product gas mainly as HBr and bromobenzene. However, by extended X-ray absorption fine structural (EXAFS) spectroscopy, Cu-O and Cu-(O) Cu species with bond distance of 1.87 and 2.95 A, respectively, were observed in the solid residues. Essentially, no Cu-Br species was found. PMID- 10665404 TI - The pH and anion effects on the heterogeneous photocatalytic degradation of o methylbenzoic acid in TiO2 aqueous suspension. AB - This investigation used UV light of 365 nm and titanium dioxide in aqueous suspension to study the photocatalytic reaction of o-methylbenzoic acid under the influence of pH values, anion additives and the varieties of titanium dioxide. From experimental results, under the condition of 5 g/l TiO2, pH 3 and light intensity of 2.45 mW/cm2, 0.1 mM of o-methylbenzoic acid could be completely decomposed in 2 h. The reaction was faster with lowering pH, and was found to be apparent first-order following Langmuir-Hinshelwood model. In the presence of anion additives, the inhibitive effect of chloride ions was larger than that of sulfate ions under acidic condition for Degussa brand titanium dioxide, but without influence using Janssen brand. Both brands, however, promoted the mineralization of o-methylbenzoic acid (o-MBA). PMID- 10665405 TI - The interaction "light, Fe(III)" as a tool for pollutant removal in aqueous solution: degradation of alcohol ethoxylates. AB - The photoinduced degradation of an alcohol ethoxylate (AE) (Brij 30) by Fe(III) in aqueous solution has been investigated. The study was carried out with the major fraction of ethoxymers having an alkyl chain length of 12 carbon atoms and n ethoxy units E (C12En). The Fe(III) sensitised degradation of this fraction occurs efficiently at 365 nm. The rate of degradation depends on the concentration of Fe(OH)2+, the most photoreactive species in terms of .OH radical formation. Formate ethoxylates were identified as photoproducts and shortening of the ethoxylated chain all along the degradation process was observed. The mechanism of Brij 30 degradation implies a major .OH radicals attack on the polyethoxylated chain. For prolonged irradiations, the total degradation of Brij 30 and of the photoproducts is obtained. Consequently, the degradation photoinduced by iron (III) could be an efficient method of AEs removal in water. PMID- 10665406 TI - Solar photocatalytic mineralization of commercial pesticides: acrinathrin. AB - A comparative study of the degradation of commercial acrinathrin spiked in water using TiO2 photocatalysis and photolysis under sunlight was performed. Samples were analysed by liquid chromatography-diode array detector (HPLC-DAD) and gas chromatography-ion trap-mass spectrometric detector (GC-ITMS). Additional total organic carbon (TOC) analyses were carried out to evaluate the mineralisation rates. One photoproduct, 2-phenoxy benzaldehyde, was unequivocally identified and evaluated by GC-ITMS during the processes. Although acrinathrin is almost destroyed when exposed to irradiation for more than 400 h, photocatalysis with TiO2 noticeably reduced degradation to a few hours. In this case, with the additional presence of peroxydisulphate, in less than 2 h acrinathrin is completely destroyed. Mineralisation of acrinathrin, without catalyst, was only around 50% after 400 h of irradiation. PMID- 10665407 TI - Light-induced disappearance of nitrite in the presence of iron (III). AB - Understanding of rapid disappearance of nitrite in natural waters and its impact on nitrogen natural cycling has remained limited. We found that NO2- disappeared rapidly in pH 3.2 aqueous Fe(III) solutions both in sunlight and in 356 nm light. Quantum yields of the NO2- loss at 356 nm were 0.049-0.14 for initial levels of 10-80 microns NO2- and 200 microns Fe(III). The NO2- loss (at 356 nm) followed apparent first-order kinetics. The rate constants were 1.3 x 10(-3) (40 microns NO2-) and 4.1 x 10(-4) s-1 (80 microns NO2-) for 100 microns Fe(III), and 2.3 x 10(-3) (40 microns NO2-) and 7.5 x 10(-4) s-1 (80 microns NO2(-1)) for 200 microns Fe(III) (t1/2 = 8.7, 27.9, 5.1, and 15.3 min, respectively). The rate constants were directly proportional to [Fe(III)]0 and inversely proportional to [NO2-]0. Agreement between the rate constants obtained experimentally and those calculated mechanistically supports the hypothesis that NO2- was oxidized to NO2 by .OH radicals from photolysis of FeOH2+ complexes, and at high [NO2-]0 (e.g., 80 microns) relative to [Fe(III)]0, hydrolysis of NO2 or N2O4 to form NO3- and NO2- could be significant. This study showed that light and Fe(III)-induced oxidation of NO2- (rate = approximately 10(-1)-10(-2) microns s-1) was more rapid than its direct photolysis (rate = approximately 10(-4) microns s-1), and the photolysis could be a significant source of .OH radicals only in cases where the Fe(III) level is much lower than the NO2- level ([Fe(III)]/[NO2-] < 1/80). This study suggests that the light and Fe(III)-induced oxidation of NO2- would be one potential important pathway responsible for the rapid transformation of NO2- in acidic surface waters, especially those affected by acid-mine drainage or volcanic activities. This study also may be of interest for modeling certain acidic atmospheric water environments. PMID- 10665408 TI - Temporal ecological assessment of oil contaminated soils before and after bioremediation. AB - Ecotoxicity methods were used to assess different soil and oil combinations before, during and after laboratory bioremediation with associated hydrocarbon analysis. Heavy, medium and light crude oil (API gravity 14, 30, and 55) was spiked (ca. 5% w/w) into two sandy soils in the laboratory having organic carbon concentrations of 0.3 (Norwood) and 4.7% (Norwood/Baccto). The earthworm (Eisenia fetida) 14-d lethality assay, the modified Microbics Microtox Solid-Phase assay, and the 14-d plant seed germination and growth assays using corn, wheat and oats, were spiked and tested during a 360-d laboratory remediation. Eisenia was the most sensitive of the three methods utilized with survival increasing throughout bioremediation with fastest toxicity reduction in the high carbon Norwood/Baccto soils where LC50's were 100% or greater at the end of 90-d whereas, > 150-d were required to achieve a similar result in the low carbon soil. Analysis of the undiluted treatments with oily soil alone showed that earthworm survival was high after 90-d in all high organic carbon soils, and after eight months in the low carbon soils, except for the Norwood soil-light oil treatment, which required 360 d to achieve 100% survival. The Microtox assay was less sensitive with EC50's 100% or greater observed after 90-d in high carbon soils and after 240-d for all low carbon soils. After bioremediation, no effects on seed germination were observed, although some plant growth inhibition effects remained. There was no direct correlation between total petroleum hydrocarbon concentrations and toxicity. PMID- 10665409 TI - Semiconductor-assisted photodegradation of lignin, dye, and kraft effluent by Ag doped ZnO. AB - This work reports a preliminary study of semiconductor-assisted photochemical degradation of lignin, Remazol Brilliant Blue R and Kraft E1 paper effluent by using ZnO and Ag-doped ZnO photocatalysts. The doped semiconductor was prepared in the reaction media by photoreduction of silver nitrate. With the use of 100 mg of ZnO and 15 mg of Ag-ZnO, almost total decolorization of the dye and lignin samples in reaction times lower than 60 min were observed. Extending the photochemical reaction up to 120 min, the total organic carbon content (TOC) was reduced in 90%. For the paper effluent, a fast decolorization was obtained for relatively short reaction times. However, de TOC reduction was negligible (near of 10%) up to high reaction times (300 min). By using the Ag-ZnO photocatalyst, the toxicity of lignin and Kraft E1 effluent toward E. Coli was completely removed. For the dye, the formation of transient toxic species was observed. PMID- 10665410 TI - Semiconductor-assisted photocatalytic degradation of reactive dyes in aqueous solution. AB - This work reports the semiconductor-assisted photochemical degradation of reactive dyes. In an oxygenated-UV-ZnO system almost total decolorization of Remazol Brilliant Blue R, Remazol Black B, Reactive Blue 221 and Reactive Blue 222 was observed in reaction times of about 60 min. Extending the photochemical treatment up to 120 min, mineralization higher than 80% for all the dyes was observed. During the same period, the residual acute toxicity was significantly reduced only for Remazol Black B. A systematic optimization study carried out by factorial design showed that for the reactive dyes tested, the ZnO semiconductor exhibits a better efficiency than that observed with anatase TiO2. A synergistic effect in the coupled TiO2-ZnO system was not observed. PMID- 10665411 TI - Use of abiotic oxidative-reductive technologies for remediation of munition contaminated soil in a bioslurry reactor. AB - The possibility to clean-up TNT contaminated soil, 400 mg TNT kg-1, surrounding Nebraska Ordnance Plant's (US), below the USEPA goal of 17.2 mg TNT kg-1 using Fenton oxidation (Fe2+ + H2O2), Fe0 reduction, combined Fe0/H2O2 and CaO2 was explored at pilot scale. Treatments were performed in a 60 l airlift reactor, which was a prototype of larger commercial unit. All the treatments reduced TNT soil concentration below the required clean-up goal and in shorter time with respect to bench scale. Using 2% (w/w) Fe0, TNT soil concentration reduced below the required standard just within 4 h. No significant TNT destruction improvement was observed when 2% Fe0 (w/w soil) was combined with four sequential additions of 0.25% H2O2. Laboratory experiments with 14C-TNT indicated that most of the 14C, approximately 80%, was unextractable residue. A time greater than 24 h was required either with Fenton reagent, 8 x (80 mg Fe2+ L-1 + 0.125% H2O2) or 0.2% (w/w) CaO2. The optimal performance of Fenton reagent was obtained when the reagent was added in eight increments rather than in a single or double dose and less cumulative amount of H2O2 (0.75%) was required with respect to bench scale (1%). PMID- 10665412 TI - Enzymatic and osmoregulative alterations in white shrimp Litopenaeus vannamei exposed to pesticides. AB - Pesticide pollution in coastal ecosystems of Sinaloa, Mexico is considered to be a cause for slow growth, increase of diseases and sometimes massive mortality of shrimp. So it was necessary to develop fast techniques to detect pesticide pollution in shrimp habitats. Enzymatic and osmoregulation tests in shrimp exposed to DDT, Lindane, Chlordane, Lorsban, Gusathion, Folidol, Diazinon and Tamaron were carried out. Activity reductions from 11 to 2 units/ml in acetylcholinesterase and from 1 to 0 units/l in transaminases (GOT and GPT) were detected. Also increases in osmoregulation were observed in shrimp exposed to Folidol, Diazinon and Gusation, whereas decreases with DDT, Lindane and Lorsban at salinity 50/1000. We conclude that pesticides are causing alterations in these biochemical functions and this kind of tests represent a rapid and inexpensive method for pesticide pollution detection. PMID- 10665413 TI - Effect of certain chemicals on the reproduction of medaka (Oryzias latipes). AB - In order to understand the effects of estrogenic chemicals on fish reproduction, we exposed male medaka (Oryzias latipes) to a natural estrogen [17 beta-estradiol (17 beta-E2)] and three estrogenic chemicals [bisphenol-A, nonylphenol (NP) and di(2-ethylhexyl)phthalate (DEHP)]. After two weeks' exposure, one male medaka was kept together with two female medaka for spawning, and the number of eggs and hatchings were compared to those of a negative control group. The results indicated that exposure to 17 beta-E2 caused a significant decrease in the number of eggs and hatchings as compared to the negative control group at and above 3 nmol/l. Also, the highest concentrations of bisphenol-A and NP caused a decrease in the number of hatchings, but no decrease in hatchings was observed in DEHP treatments. In the treatment using these chemicals the decrease in egg numbers was not so much as in hatching numbers. When compared to other in vitro studies, concentrations observed to have adverse effects on reproduction in this study are generally lower. In addition, it was suggested that physical alterations, such as an induction of plasma vitellogenin, were caused at much lower concentrations than those at which a decline in reproductivity was actually induced. PMID- 10665414 TI - Accumulation pattern and biotransformation enzyme induction in rainbow trout embryos exposed to sublethal aqueous concentrations of 3,3',4,4' tetrachlorobiphenyl. AB - Accumulation pattern of 3,3',4,4'-tetrachlorobiphenyl (PCB 77) and the exposure time needed to activate the monooxygenase (EROD) and conjugation (GST) enzyme systems of fish at the advanced embryonic stage were studied. Eyed stage embryos of rainbow trout (Oncorhynchus mykiss) were exposed to sublethal doses (0, 1, 10, and 100 micrograms/l) of PCB 77. Results indicated direct accumulation of the chemical into the eggs, but the exposure time was not long enough for PCB 77 to reach constant steady state. However, at the two lowest test concentrations (1 microgram/l and 10 micrograms/l) a temporary plateau at chemical accumulation was reached at the third exposure day (185 and 1221 ng PCB/g egg w.w). At the highest concentration (100 micrograms/l) the decrease in the accumulation rate was already evident after the first day (2182 ng PCB/g egg w.w). The chemical uptake increased again at day 7 in all the exposure groups. That event could have been caused by the increased metabolic rate of the embryos in preparation for the upcoming hatching event. The microsomal CYP1A monooxygenase system (EROD) was shown to be a sensitive indicator of embryonic exposure, being induced at the low (1 microgram/l, 182 ng/g egg) PCB concentration and after a short (3 day) exposure time. The conjugation enzyme system (GST) was shown to be functioning already at the advanced embryonic stage, although no response to the studied chemical stress was detected. PMID- 10665415 TI - New PEC definitions for river basins applicable to GIS-based environmental exposure assessment. AB - By means of GREAT-ER (Geo-Referenced Regional Exposure Assessment Tool for European Rivers) aquatic chemical fate simulations can be performed for river basins. To apply the resulting digital maps with local (river stretch specific) predicted concentrations in regional aquatic exposure and risk assessment, the output has to be aggregated to a (single) value representative of exposure in the catchment. Two spatially aggregated PEC definitions are proposed for this purpose: PECinitial (unweighted aggregation of concentrations just downstream of wastewater emissions) and PECcatchment (weighted aggregation of all average stretch concentrations). These PECs were tested using simulations for two pilot study catchments (Calder and Went, UK). This confirmed the theoretical considerations which led to the definitions, and it illustrated the need for weighting to resolve scale-dependencies. PMID- 10665416 TI - Evaluation of PNEC values: extrapolation from Microtox, algae, daphnid, and fish data to HC5. AB - In order to evaluate the risk to the environment from long term exposure of any discharged substance, toxicity thresholds are estimated, and particularly the Predicted No Effect Concentration (PNEC). This concentration can be estimated by the classic assessment factor approach or by statistical methods. These are more scientifically sound but they require several (at least 5-6) chronic ecotoxicity data, implying greater cost and time. New extrapolation methods derived from the statistical concept but requiring less data have been studied. Results show that methods based on chronic data are more reliable than methods based on acute data but the improvement is quite small. Considering the costs of chronic tests compared to acute tests, approaches based on acute data are an attractive alternative. A simple regression on the mean of the acute data gives the best results. PMID- 10665417 TI - Growing snails used as sentinels to evaluate terrestrial environment contamination by trace elements. AB - Young garden snails (Helix aspersa) reared in standard conditions (aged two months, mean weight 4.6 +/- 0.5 g) set as sentinels in cages laid on the soil for four weeks, give data for biomonitoring the environmental impact of chemicals on soil ecosystems in the field. The survival and the growth of the snails are influenced by the nature of the biotope and the level of the pollutants. Assay of cadmium, copper, lead and zinc bioaccumulated in the tissues of the sentinel snails provides information on the bioavailability of metals in the environment. The encagement model, little used for terrestrial species, can be useful in monitoring (specific and global endpoints) metal pollution of the environment in reference to the trophic level of the primary consumers. Active biomonitoring is positively compared with the passive biomonitoring. PMID- 10665418 TI - Atrazine induction of cytochrome P450 in Chironomus tentans larvae. AB - Cytochrome P450-dependent aldrin epoxidation was characterized in third instar larvae of the aquatic midge, Chironomus tentans. Optimal in vitro assay conditions for the epoxidase were pH 7.6 and 31 degrees C. Activity was linear up to 40 min of incubation time and 0.5 mg microsomal protein per incubation. The activity was concentrated in the microsomal fraction of whole body homogenates and was NADPH-dependent. The effect of atrazine exposure on aldrin epoxidase was measured to determine if this herbicide induces cytochrome P450-dependent activity. Comparisons of control and atrazine-exposed midges indicated increased epoxidase activity as a result of atrazine exposure, and a 45 kDa protein of increased intensity was observed after SDS-PAGE of microsomal protein. The molecular weight of this protein was similar in size to cytochrome P450 enzymes reported for other insects. Heme staining of SDS-PAGE gels and immunochemical studies using a Drosophila melanogaster anti-P450 polyclonal antiserum, further support the cytochrome P450 nature of this inducible 45 kDa protein. PMID- 10665419 TI - PAHs analysis of fish whole gall bladders and livers from the Natural Reserve of Camargue by GC/MS. AB - Traces of parent polycyclic aromatic hydrocarbons (PAH) in fish whole gall bladders and livers from the Natural Reserve of Camargue were determined from three different species: cels, goldfishes and catfishes by capillary gas chromatography-mass spectrometry (GC/MS) after Soxhlet extraction and florisil column cleanup. Results have been successfully correlated with biological fish parameters in order to identify adequate biomarkers of PAHs contamination. PMID- 10665420 TI - Piperonyl butoxide potentiates the synaptosome ATPase inhibiting effect of pyrethrin. AB - Pyrethrins are widely used insecticides in both agriculture and households. In many commercial formulations piperonyl butoxide (PBO) is used with pyrethrins. PBO is a well-known synergist of pyrethrins, used to intensify their effects. One of the cellular targets of pyrethrins is the sodium channel in the membrane. In the present study, the activity of the membrane-bound integral protein ATPase was studied as a biomarker for the membrane effects of pyrethrin and PBO. Cerebral synaptosomes of rat brain were used in the study. The isolation of synaptosomes was performed with the Percoll gradient method. Both total ATPase and Mg2+ activated ATPase were studied by determining inorganic phosphate. Exposure to 0.1 1000 microM of pyrethrin and to 0.4-4000 microM of PBO decreased ATPase activity dose-dependently. The most efficient mixture was the one consisting of one part of pyrethrin and four parts of PBO. The activity of total ATPase decreased 15% in concentrations of 0.1-10 microM pyrethrin, and a 50% decrease was found at 100 microM pyrethrin. The mixture of pyrethrin and PBO caused a 15-60% decrease in the total ATPase activity at 0.1-10 microM pyrethrin and 0.4-40 microM PBO. A 85% decrease was found after exposure to the mixture of 100 microM pyrethrin and 400 microM PBO. PBO alone had no effect at 0.4-40 microM concentrations, but a marked effect was seen at over 40 microM concentrations. The results indicate that PBO is an effective synergist of pyrethrin and that it is very toxic in high concentrations. The results also confirm that neuronal sodium homeostasis is one target of the neurotoxic effect of pyrethroid compounds. PMID- 10665421 TI - Cytogenetic changes in subjects occupationally exposed to benzene. AB - Humans are exposed to benzene from various occupational and environmental sources. The genotoxic effects of benzene were assessed in peripheral blood lymphocytes of 36 workers employed in the shoe industry for a period extending from seven months to over 30 years. Chromosomal aberrations and sister chromatide exchanges were used as indicators of genotoxic effects. The incidence of dicentric chromosomes in the exposed group was significantly higher than in the control group (P < 0.05). No significant increase was detected between the working period in the exposed group and chromosomal aberrations. Sister chromatide exchange (SCE) frequency was not significantly increased in the exposed group. PMID- 10665423 TI - The effect of EDTA and fulvic acid on Cd, Zn, and Cu toxicity to a bioluminescent construct (pUCD607) of Escherichia coli. AB - The hypothesis, that metal toxicity is dominated by free ion activity, was tested by comparing calculated metal activities with measured toxic responses to a genetically modified, luminescent bacterium, Escherichia coli. The toxicity of Cd, Cu, and Zn sulphate salts in the presence of EDTA and fulvic acid in well defined solutions was measured. Good agreement between free metal activity and toxicity was found for Cu but not for Zn and Cd. The toxicity relationships were altered by glucose addition to the organism. Stable chloride complexes may have contributed to the toxicity of Cd under the test conditions. The results suggest that there is not always a simple relationship between toxicity and free-ion metal concentration and that further account should be taken of competitive interactions between living cells and ligands and the physiological status of the organism. PMID- 10665422 TI - Biochemical responses of the mycorrhizae in Pinus massoniana to combined effects of Al, Ca and low pH. AB - Biochemical responses of Pinus massoniana, with and without the inoculation mycorrhizal fungus Pisolithus tinctorius at the root, to artificial acid rain (pH 2.0) and various Ca/Al ratios were investigated. Some enzymes associated with the nutritive metabolism, such as acid phosphatase, alkaline phosphatase, nitrate reductase, mannitol dehydrogenase and trehalase, in the roots, stems and leaves of plant were obviously inhibited by the artificial acid rain and Al. After treatment with pH 2.0 + Ca/Al (0/1 or 1/10) artificial acid rain, the protein content in the organs was decreased. However, the activities of superoxide dismutase (SOD) and peroxidase (POD) and glutathione (GSH) concentrations were induced. It demonstrated that acid rain and Al could induce oxygen radicals in plant. Compared with the treatments with lower pH or Al, respectively, the combination of lower pH and Al concentration was more toxic to P. massoniana. Al toxicity could be ameliorated by the addition of Ca and the amelioration was the most when the ratio was 1/1 among the various Ca/Al ratio. Infection with mycorrhizal fungus P. tinctorius at the root of P. massoniana increased the ability of the plant to resist the toxicity of artificial acid rain and Al stress. PMID- 10665424 TI - Contribution of membrane surface charge in the interaction of lead and tin derivatives with model lipid membrane. AB - The objective of this study was to determine the interaction of modified bilayer lipid membranes (BLMs) with lead and tin organoderivatives. The relative depolarization, of the lipid membrane, caused by organo metals, was used to estimate their activity. Dodecyltrimethylammonium bromide (TMDA) and sodium dodecylsulfonate (AS-12) were used to modify BLMs. The trialkylderivatives of tin (IV) were found to be the most active towards the lipid membrane, whereas dialkyltin (IV) and trialkyllead (IV) derivatives were less active. Also, a correlation exists between depolarization activity and the lipophilicity of organo metal hydrolysis products. The surface charge of modified membranes had a secondary influence on depolarization efficiency of the tin and lead derivatives. PMID- 10665425 TI - Oxidation of chlorophenols with hydrogen peroxide in the presence of goethite. AB - The use of goethite (alpha-FeOOH) and hydrogen peroxide was recently found that they could effectively oxidize organic compounds. The study was to investigate the effect of goethite particle size, goethite concentration, Fe2+ and Fe3+ on the 2-chlorophenol oxidation. Results indicated that 2-chlorophenol can be decomposed with hydrogen peroxide catalyzed by goethite and the oxidation rate increased with decreasing goethite particle size. 2-Chlorophenol degradation was almost retarded with 0.8 g/l of goethite because ferrous ions could not be produced at this condition. Addition of Fe2+ and Fe3+ can enhance the catalytic oxidation rate of 2-chlorophenol very efficiently. In conclusion, the main mechanism of goethite catalyzing hydrogen peroxide to oxidize 2-chlorophenol may be due to the catalysis of ferrous ions and goethite surface. PMID- 10665426 TI - Prediction for thermodynamic function of dioxins for gas phase using semi empirical molecular orbital method with PM3 Hamiltonian. AB - In this investigation, respective thermodynamic parameters of heats of formation, standard entropy and specific heat capacity at constant pressure for PCDDs, PCDFs, Co-PCB and PCBs as well as polychlorinated-benzenes and polychlorinated phenols have been evaluated by quantum chemical calculation using a semi empirical molecular orbital method with the PM3 Hamiltonian and statistical thermodynamic correlation. PMID- 10665427 TI - Uptake rates of semipermeable membrane devices (SPMDs) for PCDDs, PCDFs and PCBs in water and sediment. AB - Uptake rates of several PCDDs, PCDFs and PCBs were measured for semipermeable membrane devices (SPMDs) under controlled conditions in bulk water and sediment. The study was performed at 19 degrees C and 11 degrees C, and water and sediment concentrations were measured during the exposure. Linear uptake rates for specific PCDD/Fs and PCBs in 19 degrees C water varied from 34 to 111 l/m2 day and in 11 degrees C water from 8.8 to 96 l/m2 day for the whole SPMD. Uptake rates at 19 degrees C sediment ranged from 9.0 to 80 mgOC/m2 day and in 11 degrees C sediment, from 3.0 to 31, mgOC/m2 day. Partitioning of the compounds between membrane and lipid was also measured during the linear uptake phase. The membrane-lipid concentration ratios ranged from 0.02 to 1.11 depending on the compound, temperature, and bulk medium. PMID- 10665428 TI - The effects of temperature and oxygen content on the PCDD/PCDFs formation in MSW fly ash. AB - In this study, the effects of the temperature, oxygen content in the gas stream and carbon content in ash particles on PCDD/Fs formation on the fly ash surface were investigated. The optimum temperatures for dioxin formation were found at 350 degrees C for boiler ash, 300 degrees C for cyclone ash and 250 degrees C for ESP ash, respectively. Preliminary results indicate that the optimum temperature will decrease as the particle size decreases. When the O2 concentration is varied between 0% and 100%, the optimum oxygen content for PCDD/Fs formation is found to be at 7.5% for cyclone ash, and the PCDD/PCDF ratio increases with the increase of oxygen content. Dioxin formation is observed even for the gas containing no oxygen passed through the fly ash. Hence, other reacted routes that do not need O2 for dioxin formation take place on fly ash. The carbon content in fly ash is varied between 0% and 20% in this study, and the results have indicated that the maximum dioxin formation is to be found at 5%. The precursors are not injected into the fly ash or gas stream in all formation experiments, however, dioxin is still formed in fly ash. Consequently, other chlorinated routes besides Deacon reactions may take place on the fly ash surface. PMID- 10665429 TI - Estimation of dioxin emission from fires in chemicals. AB - The formation of the 17 toxic 2,3,7,8-substituted PCDDs and PCDFs during combustion of selected chemicals were measured by high-resolution GC/MS. The 16 chemicals studied were commonly used chlorinated pesticides, industrial chemicals, and PVC. In a series of experiments carried out in a DIN 53,436 furnace, 2.5 g of these compounds were burned at 500 degrees C and 900 degrees C, respectively. The resultant yields ranged from 740 ng ITEQ/g for pentachlorophenol, to below 0.01 ng ITEQ/g for PVC and dichlobenil. The results show that some chemicals generate PCDD/F in very high--possibly dangerous- amounts during burning, whereas others generate insignificant amounts. The influence of scale were studied for chlorobenzene and 4-chloro-3-nitro-benzoic acid in additional experiments, carried out in a cone calorimeter burning 20 g substance, and in ISO 9705 room test burning about 50 kg. A good agreement between the results for large and small scale indicated that formation of PCCD/F during a fire may be estimated from laboratory experiments. This suggest laboratory test may be used to screen for chemicals posing a hazard for release of PCDD/F during fires. PMID- 10665430 TI - Assessment of human health risk of dioxins in Japan. AB - The human health risk of dioxins was evaluated for four Japanese receptor groups: the general population, local residents living near a municipal solid waste incinerator, heavy fish consumers, and their infants and fetuses. In describing the risk for these groups, four endpoints, namely, cancer, reproductive dysfunction, endometriosis and neurobehavioral effect, were considered, and the incremental cancer risk and margin of exposure (MOE) corresponding to these endpoints were calculated, based on three measures of dosimetry; average daily intake, area under the curve, and body burden. The uncertainties of these risk descriptors were also evaluated by probabilistic analysis. Although the estimated risk of cancer and reproductive dysfunction were not exceptionally high in the three adult receptor groups, the MOE values for endometriosis were not sufficiently high to guarantee safety against this endpoint. Furthermore, the MOE values for neurobehavioral effects on infants and fetuses suggest that dioxins may cause a considerable risk to those of local residents and heavy fish consumers. PMID- 10665431 TI - Extraction of polychlorinated dibenzo-p-dioxins and dibenzofurans from solid samples using the Randall technique. AB - This study deals with a new sample extraction technique which minimizes pollution in analytical laboratories whilst reducing sample preparation time and cost. The device uses the Randall technique for solid sample extraction, performed by immersion of the sample in boiling solvent. The fast solubilization operated by the hot solvent leads to a sharp reduction in extraction time. This method was tested for the extraction of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) from different solid matrices using toluene as extractant, and compared with the conventional Soxhlet. The reduction in the extraction times (from 48 to 2 h) with an efficiency similar to or higher than that afforded by the conventional Soxhlet technique indicates the suitability of this method. PMID- 10665432 TI - On the possible role of acetylene in gas-phase dioxin formation. AB - The slow combustion of benzene/phenol gives rise to dibenzofuran (DBF) as major product of incomplete combustion, with negligible proportions of dibenzo-p-dioxin (DBD), or benzofuran (BF). Contrary to a recent proposal that acetylene growth reactions, e.g. BF-->DBF, are important in dioxin formation, co-combustion of benzene/phenol with acetylene--around 550 degrees C--did not alter this product pattern. Also, BF was identified as a product from degradation of DBF. PMID- 10665433 TI - Studies of toxaphene in technical standard and extracts of background air samples (Point Petre, Ontario) using multidimensional gas chromatography-electron capture detection (MDGC-ECD). AB - MDGC-ECD procedures have been used to provide insight into the compositional complexity of some of the specific peaks or clusters observed in the gas chromatographic analysis of a technical toxaphene standard, with reference to individual toxaphene congeners (Parlar # components) that are flow commercially available. These investigations have focussed initially upon those peaks and clusters recently identified (Shoeib. M., Brice, K.A., Hoff, R., 1999. Chemosphere 39, 849-871) as dominant constituents of background ambient air. Multiple electron-capturing components have been found to be present in all the species studied: the available individual toxaphene congeners have been matched against these components where possible. In similar fashion, the responses obtained in equivalent gas chromatographic elution windows from the analysis of typical processed air sample extracts have been investigated, with the results showing clear differences relative to the patterns found in the technical toxaphene standard. In most cases, the air sample shows reduced complexity with fewer components present in the cluster. Also, the presence of interfering responses (due to PCBs and other organochlorines) is quite apparent and significant, showing that major problems and errors could arise when using single column GC-ECD procedures for quantitation of toxaphene in environmental samples. The presence of certain of the Parlar species in the air samples has been confirmed and in most cases these represent the dominant toxaphene component found in the targeted cluster. Furthermore, the persistence of certain congeners in the atmospheric samples appears to be strongly dependent upon chemical structure, since the congeners in question possess an alternating exo-endo chlorine substitution pattern around the six-membered ring in the bornane skeleton. Such persistence is probably the result of lower metabolization of toxaphene residues in soils, water and sediments leading to a similar pattern in the atmosphere following volatilization. PMID- 10665434 TI - Application of GC-MS/MS for the analysis of PCDD/Fs in sewage effluents. AB - The application of high resolution gas chromatography in combination with tandem mass spectrometry in an ion trap was tested to substitute the expensive high resolution mass spectrometry in analysing polychlorinated dibenzo-p-dioxins and furans in sewage effluents. In tandem mass spectrometry, a set of parameters has to be optimised in order to attain the required sensitivity. To the best of our knowledge, this is the first time a method development for analysing PCDD/Fs with GC-MS/MS in an ion trap is described in this detail. Nine parameters are varied, including isolation window, collision induced dissociation (CID) amplitude, CID time, acquisition mass range, broadband amplitude, CID bandwidth, modulation range, filament current and ion trap temperature. This technique can be adapted to other analytes. By this optimisation, limits of detection of 0.01-0.05 ng/l are obtained. With its selectivity and sensitivity, tandem mass spectrometry is a powerful tool for the determination of PCDD/Fs in water samples. 55 sewage effluent samples from Germany were analysed. PMID- 10665435 TI - The concentration and distribution of 2,3,7,8-dibenzo-p-dioxins/-furans in chickens. AB - The concentrations of the 2,3,7,8-Cl substituted dibenzo-p-dioxins/-furans (PCDDs/PCDFs) were determined in the edible tissues of whole chicken fryers and compared with the values found in their abdominal fat. The values are presented both on a whole weight basis and on a lipid adjusted basis for each tissue. While there is a marked difference in the concentration of the 2,3,7,8-dibenzo-p dioxins in the edible tissues expressed on a whole weight basis, the lipid adjusted concentrations of the individual dioxins were not statistically different in the various tissues. This validates the use of lipid adjusted concentrations of 2,3,7,8-PCDDs/PCDFs in abdominal fat for the determination of the presence of these compounds in different tissues. PMID- 10665436 TI - PCDD/PCDF emissions from small firing systems in households. AB - This report presents results of emission measurements of polychlorinated dibenzo p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) in the flue gas of seven oil, nine gas, and two wood firing systems under laboratory conditions. The burn rate of the combustion was in the range of the rated useful heat output. Two additional test series varied the amount of combustion air and thus the heat output. The PCDD/PCDF emissions for oil- and gas-fired boilers are in the range of 0.0020-0.0142 ng I-TEQ/m3 (referring to 3% O2 in the dry flue gas). No correlation between the combustion technique and the PCDD/PCDF emissions could be established. In the tests with the wood-fired furnaces PCDD/PCDF concentrations in the flue gas ranging from 0.014 to 0.076 ng I-TEQ/m3 (referring to 13% O2 in the dry flue gas) were found. A significant correlation between the firing rate of the heating insert and the measured PCDD/PCDF concentrations was found. On examination of three typical 2,3,7,8-CDD/CDF congener profiles, a comparable pattern could be observed with natural gas and light fuel oil. The congener distribution for wood combustion is considerably different. PMID- 10665437 TI - Sorption-desorption behaviour of 2,4-dichlorophenol by marine sediments. AB - Batch kinetic and isotherm experiments were conducted to determine the sorption desorption behavior of 2,4-dichlorophenol from seawater solutions by marine sediments containing various amounts of organic carbon (from 1.02% to 12.72% dry weight). The results indicated linear type isotherms for sorption and desorption in all marine sediments studied. The observed difference in linear sorption coefficients between sorption and desorption was indicative of sorption hysteresis. The kinetic experiments showed that equilibrium was established in less than 20 h. The study is significant with respect to sediment remediation in contaminated harbors and coastal areas. PMID- 10665438 TI - Plant-promoted pyrene degradation in soil. AB - A study was conducted to investigate the capability of nine plant species to promote the degradation of pyrene in soil. The test method allowed for analysis of the entire sample of soil. More pyrene was degraded in the presence of roots of all nine species than in unplanted soil. Within approximately 8 weeks, as much as 74% of the pyrene disappeared from vegetated soil compared to 40% or less from unplanted soil. The data suggest that some of the test species may be especially useful for phytoremediation of soils contaminated with PAHs. PMID- 10665439 TI - Accumulation of phenanthrene and pyrene in rhizosphere soil. AB - A study was conducted to determine PAH concentrations in the rhizosphere of plants grown in soil containing phenanthrene or pyrene. The rhizosphere of tall fescue and wheat grown in sterile soil contained 4-5-fold higher pyrene concentrations than unplanted soil. The rhizosphere of several plant species grown in non-sterile soil temporarily contained appreciably more phenanthrene or pyrene than unplanted soil, but those PAHs were degraded with time. The data suggest that plants accumulate such hydrophobic compounds in the rhizosphere after facilitating their transport toward the roots. PMID- 10665440 TI - Mineralization of organic micropollutants (homologous alcohols and phenols) in water by vacuum-UV-oxidation (H2O-VUV) with an incoherent xenon-excimer lamp at 172 nm. AB - Twenty different organic micropollutants (for example C1-C8 homologous alcohols and some phenols) in aqueous solution were subjected to vacuum-UV-oxidation (H2O VUV) within a xenon-excimer flow-through photoreactor. The incoherent xenon excimer lamp used exhibited an emission maximum at 172 nm. At this wavelength water is efficiently photolyzed with formation of hydrogen atoms and of highly oxidative hydroxyl radicals. The short lived hydroxyl radicals initiate a series of complex interrelated radical reactions that lead finally to the mineralization of the organic material dissolved in water. The efficiency of the total organic carbon (TOC) diminution is dependent on the nature of the organic substrate treated. For example, in the series of homologous alcohols the substrate methanol is mineralized faster than 1-octanol by a factor of 2.7 under identical conditions of VUV treatment and with similar initial TOC content of the water ranging between 40 and 50 ppm. PMID- 10665441 TI - Assessment of the bioavailability of rare earth elements in soils by chemical fractionation and multiple regression analysis. AB - The bioavailability of rare earth elements (REEs) in soils was evaluated, based on the combination of chemical fractionation and multiple regression analysis. REEs in soils were partitioned by a sequential extraction procedure into water soluble (F(ws)), exchangeable (F(ec)), bound to carbonates (F(cb)), bound to organic matter (F(om)), bound to Fe-Mn oxides (F(fm)) and residual (F(rd)) fractions. Alfalfa (Medicago Staiva Linn.) had been grown on the soils in a pot culture experiment under greenhouse conditions for 35 days. The concentrations of REEs in fractions and plant were determined by inductively coupled plasma-mass spectrometry (ICP-MS). Chemical fractionation showed that (F(ws)) fraction of REEs was less than 0.1% and residual (F(rd)) was the dominant form, more than 60% in soils. Bioaccumulation of REEs was observed in Alfalfa. REE availability to the plant was evaluated by multiple regression analysis. F(ws), F(ec), F(cb) and F(om) fractions were significantly correlated with REE uptake by alfalfa. But the exchangeable Pr(F(ec)) was significantly correlated with Pr concentration in alfalfa. F(ec), F(cb) and F(om) greatly contributed to La and Nd bioavailability; F(ec) and F(om) to Ce, Gd and Dy; F(ec) and F(cb) to Yb; and F(ws), F(ec) and F(om) to total REEs. This meant that the bioavailability of different species of REEs varied with individual REE. The results of this study indicated that the sequential extraction procedure, in conjunction with multiple regression analysis, may be useful for the prediction of plant uptake of REEs from soils. PMID- 10665442 TI - Determination of selected fate and aquatic toxicity characteristics of acrylic acid and a series of acrylic esters. AB - Acrylic acid, methyl acrylate, ethyl acrylate, and butyl acrylate are commercially important and widely used materials. This paper reports the results of a series of fate and aquatic toxicity studies. The mobility in soil of acrylic acid and its esters ranged from 'medium' to 'very high'. Calculated bioconcentration factors ranged from 1 to 37, suggesting a low bioconcentration potential. Acrylic acid and methyl acrylate showed limited biodegradability in the five day biochemical oxygen demand (BOD5) test, while ethyl acrylate and butyl acrylate were degraded easily (77% and 56%, respectively). Using the OECD method 301D 28-d closed bottle test, degradability for acrylic acid was 81% at 28 days, while the acrylic esters ranged from 57% to 60%. Acrylic acid degraded rapidly to carbon dioxide in soil (t1/2 < 1 day). Toxicity tests were conducted using freshwater and marine fish, invertebrates, and algae. Acrylic acid effect concentrations for fish and invertebrates ranged from 27 to 236 mg/l. Effect concentrations (LC50 or EC50) for fish and invertebrates using methyl acrylate, ethyl acrylate, and butyl acrylate ranged from 1.1 to 8.2 mg/l. The chronic MATC for acrylic acid with Daphnia magna was 27 mg/l based on length and young produced per adult reproduction day and for ethyl acrylate was 0.29 mg/l based on both the reproductive and growth endpoints. Overall these studies show that acrylic acid and the acrylic esters studied can rapidly biodegrade, have a low potential for persistence or bioaccumulation in the environment, and have low to moderate toxicity. PMID- 10665443 TI - Analysis of volatile organic compounds, using the purge and trap injector coupled to a gas chromatograph/ion-trap mass spectrometer: review of the results in Dutch surface water of the Rhine, Meuse, Northern Delta Area and Westerscheldt, over the period 1992-1997. AB - Volatile organic compounds are widely introduced into the Dutch aquatic environment. Liquid-liquid extraction and isolation by means of resins give poor recoveries for volatiles. In this study, a method has been developed to analyse these compounds with a purge and trap injection (PTI) coupled to a chromatograph/ion trap mass spectrometer. Volatile compounds are "purged" from the sample by carrier gas flow and transported through a condenser to a cooled trap. The bulk of liquid matrix is condensed in the condenser, while volatile compounds are left unaffected. The compounds of interest are trapped at a low temperature, by liquid nitrogen in the cooled trap of fused silica. Injection takes place by flash heating of the trap. The detection limits of the volatile compounds are in the range of 0.001-0.04 microgram l-1, in full spectrum mode. In this paper a review of the results in Dutch surface water of the Rhine, Meuse, Northern Delta Area and Westerscheldt, over the period 1992-1997, is presented. For calamities causing high levels of volatiles, the method is very useful. The compounds can be monitored over a certain period. In the Meuse, high levels of volatile organic compounds are observed. Diisopropyl ether at a maximum of 592 micrograms l-1. PMID- 10665444 TI - Adsorption/thermal desorption-GC/MS for the analysis of pesticides in the atmosphere. AB - An analytical methodology using Automatic Thermal Desorption (ATD) and GC/MS was developed for the determination of the pesticides alachlor, atrazine, captan, formothion, lindane and phosalone in atmospheric samples. This methodology was developed to evaluate the atmospheric contamination by pesticides during treatments and by post-application. Atmospheric samples were collected by using (4 i.d. x 89 mm) stainless steel sampling tubes containing 125 mg of adsorbents at a flow rate of 80 ml min-1. Different types of adsorbents were tested for their ability to efficiently trap pesticides under study: Tenax TA, Carbopack Y, Carbopack B, Carbotrap, Carboxen, Chromosorb 106 and XAD-4. Results of experiment show that Tenax gives the better results for all the pesticides used but the use of the thermal-desorption method, especially for pesticides with low volatility and/or poor thermal stability presents some difficulties. This method was validated by the analysis of the contamination of atmosphere, through volatilization by post-application processes, of atrazine in a parcel of 1 ha. PMID- 10665445 TI - The concentration, apparent molecular weight and chemical reactivity of silica from groundwater in southern Nevada. AB - Sorption of radionuclides, metals and organic compounds to colloidal particles has been suggested to increase the mobility of these pollutants in groundwater. Because silicates and alumino-silicates can be important components of groundwater colloids, we have conducted a study to characterize the nature of silica in various springs and wells in Southern Nevada and to determine the extent that silica may be associated with colloidal particles that can participate in pollutant transport. The total silica content was measured using inductively coupled plasma emission spectroscopy (ICP). In addition, reactive silica was measured using the silica molybdate colorimetric technique. The apparent molecular weight of the silica was investigated using split-flow-lateral transport-thin-cell-fractionation (SPLITT) which can readily distinguish between colloidal and low molecular weight associations. This study indicates that silica does not tend to form stable inorganic colloids in Southern Nevada groundwaters but exists as low molecular weight species. However, water from one of the test facilities on the Nevada Test Site (NTS) did contain stable siliceous colloids that could have important implications for the modeling the transport of radionuclides at this site. PMID- 10665446 TI - Metal contents of fish from cultureponds near scrap metal reclamation facilities. AB - Milkfish (Chanos chanos) from four fish-culture ponds adjacent to different metal recovery facilities along the Er-Jen River area, Tainan, Taiwan, were sampled to determine their metal contents. Chemical analysis showed that fish tissue contained different concentrations of Cu: 0.71-6.37 micrograms/g, Pb: ND (not detectable) approximately 41.04 micrograms/g, Cd: ND approximately 0.41 microgram/g, Al: 6.75-64.11 micrograms/g, Ni: 0.062-0.504 microgram/g and Zn: 16.11-41.86 micrograms/g. The average concentrations of Cu, Al, Zn, Cd and Pb in fish samples from some of the ponds were significantly higher than those from the reference pond. In addition, there were variations in metal concentrations of fish collected from different ponds. Pond D had the highest mean values of Cu, Cd and Zn, and Pond B of Al and Pb. Further investigations are needed to determine the source of metal contamination in the fish. PMID- 10665447 TI - Determination of nitrate and other water quality parameters in groundwater from UV/Vis spectra employing partial least squares regression. AB - The use of UV/Vis spectroscopy in combination with partial least squares (PLS) regression for the simultaneous prediction of nitrate and non-purgeable organic carbon (NPOC) in groundwaters was evaluated. A model of high quality was obtained using first order derivative spectra in the range 200-300 nm. Inclusion of non-UV absorbing constituents in the modeling procedure, i.e., chloride, sulfate, fluoride, total carbon (TC), inorganic carbon (IC), alkalinity, pH and conductivity was also evaluated. This model seemed to be useful for prediction of chloride, TC, IC, alkalinity and conductivity, while its ability to predict sulfate, fluoride and pH was poor. In conclusion, application of PLS regression, which requires neither filtration of samples nor addition of chemicals, is a promising alternative for fast interpretation of geochemical patterns of groundwater quality. PMID- 10665448 TI - Prediction of pesticide concentrations in the atmosphere using an atmospheric diffusion model (linear source plume model). AB - The foundational model to predict concentration of pesticides in the atmosphere outside of the sprayed area was developed using the results of measured concentration in the atmosphere, in reference to the atmospheric diffusion model utilized for the air pollution prediction model. The atmospheric diffusion model assumes that the applied area was a topographically flat farmland, that wind direction and wind speeds were constant, and the pesticide was constantly discharged from the emission line sources. Therefore the linear source plume model (LSPLM) was developed. The concentration in the atmosphere was predicted by assigning the property of the pesticides and various conditions of measurements of the model, and compared with the measured them, then the adaptability of the model was examined. As a result, the correlation between the measured value and the predicted value in paddy and forested areas was significant (P < 0.01) although deviations in the order of tens were observed, the measured value and the predicted value were generally in agreement. PMID- 10665449 TI - Fungal degradation of fluorene. AB - A selection of 30 strains of micromycetes known as good degraders of polychlorinated aromatic compounds, mostly isolated from soil and belonging to various taxonomic groups, have been investigated to degrade fluorene. Toxicity assays, first evaluated on solid media, have shown high growth inhibition at concentrations above 0.001 g l-1 only towards 23% of strains. Degradation of fluorene (0.005 g l-1) was then investigated in liquid synthetic medium for 2 days and evaluated by HPLC. Among the 30 strains tested, 12 could be considered as best degraders because of a rate of degradation at 60% or over. 3 strains of Cunninghamella genus were very efficient (mean of degradation: 96%) but different strains from Ascomycetes. Basidiomycetes and Deuteromycetes were also efficient 11 strains are not yet reported in the literature: Aspergillus terreus, Bjerkandera adusta, Ceriporiopsis subvermispora, Colletotrichum dematium, Cryphonectria parasitica, Cunninghamella blakesleeana, C. echinulata, Drechslera spicifera, Embellisia annulata, Rhizoctonia solani and Sporormiella australis. A metabolic approach with standard compounds (9-fluorenol and 9-fluorenone) indicated the presence of these monooxygenated derivatives for most of the strains. PMID- 10665450 TI - An analysis of the 'Modified Sturm Test' data. AB - The 'Modified Sturm Test' uses carbon dioxide production as the primary end point in assessing the biodegradation potential of organic chemicals. This test was conducted by a commercial laboratory to assess the potential biodegradability of an oil stabilizer sample from an oil company in Canada. There was a high percentage conversion of total organic carbon present in the sample but carbon dioxide measured was low. Many possibilities were analyzed in this paper in order to understand the situation. The analysis showed that the test was subject to criticism from the point of view of CO2 measurement, 10-day window period, and aeration/mixing conditions. PMID- 10665451 TI - Glyphosate adsorption on soils of different characteristics. Influence of copper addition. AB - Results of glyphosate (GPS) adsorption on three soils of different characteristics show that the interaction of this pesticide with the soils was not related to their CEC and clay minerals content, but to the content of iron and aluminum amorphous oxides and organic matter. The presence of Cu in treatment solutions enhanced GPS adsorption, due to several reasons: GPS coordinates strongly to Cu, and Cu GPS complexes formed seem to have higher ability to be adsorbed on the soil than free GPS; GPS adsorption can take place on sites where Cu was previously adsorbed, acting as a bridge between the soil and GPS; when Cu was present the solution pH decreased, and GPS adsorption increased, since lower pHs lead to the formation of GPS species with lower negative charge, which are adsorbed more easily on the negatively charged soil surfaces. PMID- 10665452 TI - Benzene accumulation in horticultural crops. AB - In this study apple, blackberry and cucumber crops were exposed to elevated levels of benzene under controlled conditions. Benzene was retained in fruits of all crops, but only accumulated in leaves of blackberries and apples. The retention by cucumber fruits is suggested to result from the longer pathway for the desorption of benzene as a consequence of their increased tissue depth compared to leaves. The process of accumulation in blackberry and apple leaves is unknown. The ingestion of benzene via the food-chain pathway on the basis of this study is concluded not to be significant. PMID- 10665453 TI - Photocatalytic degradation of lignin and lignin models, using titanium dioxide: the role of the hydroxyl radical. AB - The role of hydroxyl radicals on the degradation of lignins during a cellulosic pulp bleaching process including a photocatalytic stage, was assessed using peroxyformic acid lignins EL1 and REL1 and two phenolic niphenyl lignin models 1 and 2. The irradiations were performed in the absence of photocatalyst TiO2 and H2O2 (condition a), in the presence of TiO2 (condition b) and in the presence of H2O2 (condition c). The experiments were conducted in alkaline (pH approximately 11) aqueous ethanol solutions with oxygen bubbling. The relative phenolic content of the irradiated solutions, which is indicative of the involvement of hydroxyl radicals, was determined by ionization absorption spectroscopy. The results obtained show that the catalyzed reaction involves both degradation of the phenolate groups by electron transfer and hydroxylation of the lignin aromatic structure. Benzyl alcohol structural elements in sodium borohydride reduced lignin REL1 and compound 2 were also found as good trapping agents for the hydroxyl radicals. The degradation of EL1 was studied by measuring its fluorescence emission by comparison to the fluorescence of compound 2. The emission spectra indicate that some biphenyl phenolate anions in EL1 are reacting under UV/visible irradiation and some others, probably polyphenolic chromophores emitting less fluorescence, are formed. PMID- 10665454 TI - Outcomes in a referral cohort of patients with anxiety disorders. AB - This study describes 6- and 12-month outcomes in a referral cohort with anxiety disorders and identifies treatment and prognostic factors related to these outcomes. Patients were recruited at three general hospital clinics, two psychologist-run clinics, and one psychiatric hospital clinic. Outcomes included severity of symptoms, physical and mental health status, and subjective global change in problem severity. Of 254 patients eligible for follow-up, 165 (65.0%) completed a follow-up questionnaire. Methods of treatment included consultation with return to the primary care physician (38.2%); or continued treatment at the clinic, with medications (16.4%), psychotherapy (22.4%), or both (23.0%). Both severity of symptoms and mental health status improved but remained abnormal at follow-up. In multiple logistic regression, subjective global improvement was related to a diagnosis of panic disorder only, treatment with psychotherapy, and type of referral. Change over time in symptom severity was related to clinic type, and change over time in mental health was related to clinic type and duration of previous treatment. PMID- 10665455 TI - Affect regulation in alexithymia: an ethological study of displacement behavior during psychiatric interviews. AB - Displacement activities (i.e., self-directed behaviors such as self-touching, scratching, and self-grooming) are a reliable ethological indicator of increased emotional and physiological arousal throughout the phylogenetic scale. We hypothesized that, in alexithymic individuals, the failure to regulate cognitively distressing emotions might result in increased displacement behavior. The nonverbal behavior of 30 patients with depressive or anxiety disorders was video-recorded during psychiatric interviews and analyzed using an ethological scoring system. Before being interviewed, each patient completed the Twenty-Item Toronto Alexithymia Scale (TAS-20), the Beck Depression Inventory (BDI), and the state form of the State-Trait Anxiety Index (STAI-S). Ethological data confirmed the hypothesis of the study. The patients with more pronounced alexithymic features showed a significantly higher frequency of displacement activities during interviews. At the same time, these patients reported levels of self-rated anxiety and depression equivalent to those reported by nonalexithymic patients. Such a dissociation between cognitive appraisal of emotion and nonverbal behavior reflecting increased emotional arousal supports the view that alexithymia implies a failure to elevate emotions from a preconceptual level of organization to the conceptual level of mental representations. PMID- 10665456 TI - Parental style and vulnerability to depression: the role of core beliefs. AB - This study considers the potential role of core beliefs (unconditional schema level representations) in the relationship between recalled parenting in childhood and major depression in adulthood, comparing a group of depressed outpatients (N = 60) with a healthy community sample (N = 67). The depressed group were differentiated by poorer perceived parenting (low care and high overprotection) and by three unhealthy core beliefs (defectiveness/shame, self sacrifice, and insufficient self-control). Among nonclinical participants, it appears that vulnerability to harm beliefs act as a partial mediator of the relationship between poor paternal care and the development of depressive features. In contrast, a broader set of core beliefs appears to mediate the relationship of maternal bonding and paternal overprotection with depressive symptoms among the depressed group. The findings suggest that clinical work with adults with major depression might need to take account of parental style. Where parents are reported to be uncaring or overprotective, cognitive-behavioral therapy might need to include a schema-focused component. PMID- 10665457 TI - Risk factors associated with the dissociative experiences of borderline patients. AB - The objective of this study was to identify the risk factors associated with the dissociative symptomatology of borderline patients. The Dissociative Experiences Scale--a 28-item self-report measure that has well documented reliability and validity--was administered to 290 criteria-defined borderline patients and 72 axis II comparison subjects. Semistructured interviews pertaining to difficult childhood experiences and adult experiences of being a victim of violence were administered to these patients blind to diagnostic status. In the sample of borderline patients alone, multiple regression analyses revealed that four risk factors were found to be significantly associated with the level of dissociation reported by these 290 patients: inconsistent treatment by a caretaker, sexual abuse by a caretaker, witnessing sexual violence as a child, and adult rape history. In the combined sample of axis II patients, the borderline diagnosis joined these four "traumatic" factors as a significant predictor of the overall level of dissociation reported by these 362 personality-disordered inpatients. The results of this study suggest that both sexual trauma and something intrinsic to the borderline diagnosis itself are risk factors for dissociative phenomena among borderline patients. PMID- 10665458 TI - Personality traits in schizophrenia: comparison with a community sample. AB - The objective of this study was to compare personality trait profiles in patients with schizophrenia and healthy controls. Male outpatients with schizophrenia (N = 24) and a male nonpsychiatric community sample (N = 46) completed the NEO-FFI personality questionnaire. Multivariate analyses were used to compare mean scale scores and scale profiles for each group. The overall personality profile of clinically stable patients with schizophrenia differed significantly from that of a community sample. On individual scales, patients scored significantly higher on neuroticism and significantly lower on conscientiousness. These results confirm and extend those of previous studies that used normative data for comparison and a much longer version of the same personality questionnaire. Prospective studies of populations at risk are needed to determine whether group differences reflect a premorbid diathesis for schizophrenia or a secondary effect of serious mental illness. PMID- 10665459 TI - Research on religious variables in five major adolescent research journals: 1992 to 1996. AB - A review of quantitative research studies published between 1992 and 1996 in five major adolescent journals revealed that 11.8% (109 of 922) included a measure of religion. This percentage (11.8%) is 3 to 10 times higher than that found in previous reviews of empirical research in psychological and psychiatric journals, suggesting that adolescent research journals are more sensitive to the role of religious factors on mental health than research in related disciplines. The results are discussed in the context and philosophy of the adolescent research and in comparison with related disciplines. PMID- 10665460 TI - Medically self-harming behavior and its relationship to borderline personality symptoms and somatic preoccupation among internal medicine patients. PMID- 10665461 TI - Caregiver burden in Shy-Drager syndrome. PMID- 10665462 TI - Fluoxetine as an adjunct to conventional antipsychotic treatment of schizophrenia patients with residual symptoms. PMID- 10665463 TI - Sleep hygiene and sleep-onset insomnia. PMID- 10665464 TI - Neuropsychiatric effects of insular stroke. AB - The neuropsychiatric effects of insular damage in humans have not previously been examined. We therefore examined the neuropsychiatric impairment in seven patients with left insular stroke, six patients with right insular stroke, six patients with left hemisphere noninsular stroke, and six patients with right hemisphere noninsular stroke. Between 4 and 8 weeks after acute stroke, patients were administered a neuropsychiatric battery. Patients with right insular lesions had a greater frequency of subjective anergia and underactivity (Fisher's exact p = .002) as well as tiredness (Fisher's exact p < .002) compared with patients with non-insular lesions or left insular lesions. Subjective feelings of impaired energy or drive after right insular damage may result from disconnection between the insula and the frontal lobe or the anterior cingulate cortex, structures that have been associated with willed action and motor behavior. PMID- 10665465 TI - Dissociation in temporal lobe epilepsy and pseudo-epileptic seizure patients. AB - Patients with epileptic seizures (ES) and especially those with temporal lobe epilepsy (TLE) share many symptoms with patients with pseudo-epileptic seizures (PES), and the differentiation between them is often difficult There is growing evidence that a subgroup of PES patients suffer from a dissociative disorder. It is recognized that dissociative symptoms pertain to both psychological and somatoform components of experience. Questionnaires assessing dissociation might provide positive criteria for the diagnosis of PES. In this study, the Dissociation Questionnaire (DIS-Q) and the Somatoform Dissociation Questionnaire (SDQ-20) were administered to patients with ES (TLE, non-TLE) and PES. To control for the influence of general psychoneurotic complaints, the SCL-90 was administered. Apart from this, answers on a trauma questionnaire were related to the diagnosis. Results showed that PES patients scored significantly higher on the SDQ-20, also after correction with the SCL-90, and no difference was found on the DIS-Q. Also, PES patients significantly more often reported sexual traumatic experiences. A logistic regression revealed that results on the SDQ-20 have no independent value in addition to the contribution of gender, age, age at seizure onset, and the presence of sexual abuse in the prediction of the diagnosis. In conclusion, somatoform and not psychological dissociative symptoms are characteristic for PES patients in comparison to ES patients. Other measures are needed within the framework of the differential diagnosis between PES and ES. PMID- 10665466 TI - Assessing clinical predictions of early rehospitalization in schizophrenia. AB - This study determines patient characteristics that predict early hospital readmission in schizophrenia and evaluates the extent to which inpatient staff accurately predict these readmissions. Adult inpatients with schizophrenia or schizoaffective disorder (N = 262) were evaluated at hospital discharge and 3 months later to assess hospital readmission. At hospital discharge, inpatient staff were asked to identify which patients were likely to be readmitted during this period. Comparisons were made between patients who were or were not readmitted and between readmitted patients who were or were not identified by staff as likely to be readmitted; 24.4% of the sample were readmitted within 3 months of hospital discharge. Early readmission was associated with four or more previous hospitalizations (85.7% vs. 57.7%, p = .004), comorbid substance use disorder (60.3% vs. 35.5%, p = .0006), major depression (40.6% vs. 26.8%, p = .04), absence of a family meeting with inpatient staff (58.2% vs. 41.8%, p = .02), and prescription of a conventional rather than an atypical antipsychotic medication (93.7% vs. 83.8%, p = .045). Twelve of the 63 readmitted patients were correctly predicted by staff to re-hospitalize. Staff tended to overestimate the risk of rehospitalization in patients with a poor therapeutic alliance, low global function, or initial involuntary admission and to underestimate the risk in patients with alcohol use disorders or four or more previous psychiatric hospitalizations. Early rehospitalization is common in schizophrenia and difficult to predict. Greater emphasis on comorbid alcohol use disorders and a history of multiple previous admissions may help clinicians identify patients at greatest risk for early rehospitalization. PMID- 10665467 TI - A specific attentional bias in suicide attempters. AB - Selective attention in patients after an attempted suicide was investigated to find out whether a specific attentional bias for suicide-related materials exists and to clarify the possible role of emotions in the bias. Thirty-one patients who had previously attempted to commit suicide and 31 control participants took part in a modified Stroop task. The suicidal patients took significantly longer to name the colors of suicide-related words compared with other words, whereas color naming times of the control participants did not differ for suicide-related, neutral, positive, or negative words. Therefore, the attentional bias exhibited by suicidal patients was highly specific. There was no relation between the bias and measures of anxiety, depression, or hopelessness, whereas suicidal ideation correlated significantly with the attentional bias. PMID- 10665468 TI - Core beliefs in anorexic and bulimic women. AB - There is research evidence to suggest the presence of dysfunctional cognitions in anorexia nervosa and bulimia nervosa that are not related to food, weight, or shape. These maladaptive cognitions have not been addressed by the conventional cognitive behavioral models of etiology or therapy. This study aimed to assess the impact of unhealthy core beliefs on eating disorders and their symptoms. Twenty restricting anorexics, 10 bulimic anorexics, 27 bulimics, and 23 normal controls completed Young's Schema Questionnaire. Eating behaviors and attitudes were also measured. The results indicate that both anorexic and bulimic women had significantly higher levels of unhealthy core beliefs than comparison women, but the clinical groups only differed on one individual core belief (entitlement). However, there were different patterns of association between core beliefs and eating psychopathology in anorexic and bulimic women. It is suggested that future clinical practice should incorporate core beliefs as a potential element in the assessment and treatment of eating disorders. PMID- 10665469 TI - Lifetime severity index for cocaine use disorder (LSI-Cocaine): a predictor of treatment outcomes. AB - We developed a lifetime severity index for cocaine use disorder and examined its predictive validity of posttreatment outcome using data from the national Drug Abuse Treatment Outcome Study. The index, based on 28 items, considered frequency of use, recency, dependency, and attempt to quit. A higher value of the index, indicating greater severity, predicted a greater likelihood of relapse (the odds ratios were 5.7 for high severity and 4.4 for medium severity, relative to low severity) and shorter time to relapse. Similarly, the polytomous logistic analysis indicated that the index predicted levels of posttreatment cocaine use (odds ratios of daily use were 47.8 for the high severity and 18.8 for medium severity; the corresponding odds ratios of weekly use were 6.75 and 5.10 and for less-than-weekly use were 3.35 and 3.57). The index can be a useful measure for both clinical and research purposes. PMID- 10665470 TI - Unrecognized dissociation in psychotic outpatients and implications of ethnicity. PMID- 10665471 TI - Traumatic stress and depression in a group of plane crash survivors. PMID- 10665472 TI - Clinician response to treatment refractory panic disorder: a survey of psychiatrists. PMID- 10665473 TI - Does undergraduate education have an effect on Edinburgh medical students' attitudes to psychiatry and psychiatric patients? PMID- 10665474 TI - Inhibition of tumor growth and metastasis by targeting tumor-associated angiogenesis with antagonists to the receptors of vascular endothelial growth factor. AB - Angiogenesis, the formation of new blood vessels, is essential for both tumor growth and metastasis. Recent advances in our understanding of the molecular mechanisms underlying the angiogenesis process and its regulation have led to the discovery of a variety of pharmaceutical agents with anti-angiogenic activity. The potential application of these angiogenesis inhibitors is currently under intense clinical and pre-clinical investigation. Compelling evidence suggests that vascular endothelial growth factor (VEGF) and its receptors play critical roles in tumor-associated angiogenesis, and that they represent good targets for therapeutic intervention. This has been demonstrated in a variety of animal tumor models in which disabling the function of VEGF and its receptors was shown to inhibit both tumor growth and metastasis. We have produced a panel of antibodies directed against the VEGF receptor 2, KDR/F1k-1. These antibodies potently block VEGF/KDR/F1k-1 interaction, and inhibit VEGF-stimulated activation of the receptor and proliferation of human endothelial cells. Further, the antibodies significantly inhibited tumor-associated angiogenesis in several animal models. Antagonists of VEGF and/or its receptors may offer higher specificity towards tumors with reduced side effects, and may be less likely to elicit drug resistance compared to conventional therapy. Anti-angiogenesis therapy represents a novel strategy for the treatment of cancer and other human disorders where pathological angiogenesis is involved. PMID- 10665475 TI - Tyrosine kinase inhibitors in preclinical development. AB - Due to the limited efficacy of cytotoxic chemotherapy in the treatment of advanced malignancy and its excessive toxicity precluding its use in chemoprevention, new therapeutic and preventive strategies have been sought. One of the most interesting of these new approaches is the manipulation of signal transduction pathways. Among the approaches being considered to eventuate such a strategy is the inhibition of autophosphorylation, a critical first step in the signal transduction pathways of many cell surface receptor tyrosine kinases, as well as of non-receptor tyrosine kinases. This article is intended to review those tyrosine kinase inhibitors that are currently in preclinical development, for which there are data to support consideration for their use in chemoprevention or cancer treatment. We will focus upon those agents that have received attention in the past several years. PMID- 10665477 TI - Farnesyl protein transferase inhibitors and other therapies targeting the Ras signal transduction pathway. AB - The year 2000 will be a significant date for the field of Ras-related therapies since numerous agents will have Phase II clinical efficacy data maturing to provide proof of principle for this cancer treatment strategy. These data will also provide an important milestone for the cancer research community since these molecules represent a small vanguard of oncology drug discovery projects predicated on molecular targets. We can only hope that these agents are a successful harbinger for the formidable number of targeted therapies that will be entering development pipelines in the coming years. PMID- 10665478 TI - Epidermal growth factor receptor inhibition in cancer therapy: biology, rationale and preliminary clinical results. AB - The epidermal growth factor receptor (EGFR), a growth factor receptor involved in the regulation of cellular differentiation and proliferation, is highly expressed by many tumor cells. In light of a relationship between overexpression of EGFR and clinically aggressive malignant disease, EGFR has emerged as a promising target for cancer therapy. In recent years, several molecular strategies have been explored to modulate either the EGFR itself, or the downstream signal beyond the cell surface receptor. One of the most promising current strategies involves the use of anti-EGFR monoclonal antibodies (mAbs), either alone or in combination with conventional cytotoxic modalities such as chemotherapy or radiotherapy. This review focuses primarily on recent progress in the development of anti-EGFR mAbs, and examines their potential in the treatment of cancer. PMID- 10665476 TI - Protein kinase C targeting in antineoplastic treatment strategies. AB - Neoplastic cell survival is governed by a balance between pro-apoptotic and anti apoptotic signals. Noteworthy among several anti-apoptotic signaling elements is the protein kinase C (PKC) isoenzyme family, which mediates a central cytoprotective effect in the regulation of cell survival. Activation of PKC, and subsequent recruitment of numerous downstream elements such as the mitogen activated protein kinase (MAPK) cascade, opposes initiation of the apoptotic cell death program by diverse cytotoxic stimuli. The understanding that the lethal actions of numerous antineoplastic agents are, in many instances, antagonized by cytoprotective signaling systems has been an important stimulus for the development of novel antineoplastic strategies. In this regard, inhibition of PKC, which has been shown to initiate apoptosis in a variety of malignant cell types, has recently been the focus of intense interest. Furthermore, there is accumulating evidence that selective targeting of PKC may prove useful in improving the therapeutic efficacy of established antineoplastic agents. Such chemosensitizing strategies can involve either (a) direct inhibition of PKC (e.g., following acute treatment with relatively specific inhibitors such as the synthetic sphingoid base analog safingol, or the novel staurosporine derivatives UCN-01 and CGP-41251) or (b) down-regulation (e.g., following chronic treatment with the non-tumor-promoting PKC activator bryostatin 1). In preclinical model systems, suppression of the cytoprotective function(s) of PKC potentiates the activity of cytotoxic agents (e.g., cytarabine) as well as ionizing radiation, and efforts to translate these findings into the clinical arena in humans are currently underway. Although the PKC-driven cytoprotective signaling systems affected by these treatments have not been definitively characterized, interference with PKC activity has been associated with loss of the mitogen activated protein kinase (MAPK) response. Accordingly, recent pre-clinical studies have demonstrated that pharmacological disruption of the primary MEK-ERK module can mimic the chemopotentiating and radiopotentiating actions of PKC inhibition and/or down-regulation. PMID- 10665479 TI - Antiandrogens in prostate cancer. AB - Antiandrogens competitively inhibit ligand binding to the androgen receptor (AR), and are used therapeutically in prostate cancer patients. The AR functions as a ligand dependent transcription factor that transduces androgen binding into increased transcription of androgen dependent genes. AR blockade induces programmed cell death in the vast majority of malignant and benign prostate cancer cells that have not previously been exposed to androgen ablation. The antiandrogens are divided structurally into the steroidal and non steroidal agents. The biological effects of the steroidal versus nonsteroidal agents are distinguished by differences in their effects on serum testosterone levels, and by their activity at receptors other than the androgen receptor. There is extensive clinical experience in the palliative and curative therapy of prostate cancer using antiandrogens as monotherapy or antiandrogens in combination with luteinizing hormone agonists or surgical castration. Prolonged therapy with antiandrogens selects for mutations in the AR that change the AR ligand specificity and permits stimulation by ligands that are usually inhibitory. These mutations give insight into one of the means by which prostate cancer progresses despite antiandrogen therapy, and also helps to explain the antiandrogen withdrawal syndrome. Areas of active research that may affect the future use of antiandrogens include the ongoing evaluation of antiandrogens in combination with 5 alpha reductase inhibitors to achieve AR blockade without inducing castrate testosterone levels. There is also interest in developing selective androgen receptor modulators (SARM) that can achieve AR blockade without causing the increased testosterone levels produced by the nonsteroidal antiandrogens currently in use. PMID- 10665481 TI - Flavopiridol: the first cyclin-dependent kinase inhibitor in human clinical trials. AB - The discovery and cloning of the cyclin-dependent kinases (cdks), main regulators of cell cycle progression, allowed several investigators to design novel modulators of cdk activity. Flavopiridol (HMR 1275, L86-8275), a flavonoid derived from an indigenous plant from India, demonstrated potent and specific in vitro inhibition of all cdks tested (cdks 1, 2, 4 and 7) with clear block in cell cycle progression at the G1/S and G2/M boundaries. Moreover, preclinical studies demonstrated the capacity of flavopiridol to induce programmed cell death, promote differentiation, inhibit angiogenic processes and modulate transcriptional events. The relationship between the latter effects and cdk inhibition is still unclear. Initial testing in early clinical human trials with infusional flavopiridol showed activity in some patients with non-Hodgkin's lymphoma, renal, prostate, colon and gastric carcinomas. Main side effects were secretory diarrhea and a pro-inflammatory syndrome associated with hypotension. Biologically active plasma concentrations of flavopiridol (approximately 300-500 nM) are easily achievable in patients receiving infusional flavopiridol. Phase 2 trials with infusional flavopiridol in several tumor types, other schedules and combination with standard chemotherapies are being assessed. In conclusion, flavopiridol is the first cdk inhibitor to be tested in clinical trials. Although important questions remain to be answered, this positive experience will stimulate the development of novel cdk modulators for cancer therapy. PMID- 10665482 TI - Age-related white matter changes and cognitive impairment. PMID- 10665483 TI - Congenital muscular dystrophy: an expanding clinical syndrome. PMID- 10665480 TI - Antiestrogens--tamoxifen, SERMs and beyond. AB - Estrogens play a central role in reproductive physiology. The cellular effects of estrogens are mediated by binding to nuclear receptors (ER) which activate transcription of genes involved in cellular growth control. At least two such receptors, designated ERalpha and ERbeta, mediate these effects in conjunction with a number of coactivators. These receptors can directly interact with other members of the steroid receptor superfamily. A complex cross-talk exists between the estrogen-signaling pathways and the downstream signaling events initiated by growth factors, such as epidermal growth factor and insulin-like growth factors. Estrogens are also a causative factor in the pathogenesis of a variety of neoplastic and non-neoplastic diseases, including breast cancer, endometrial cancer, endometriosis, and uterine fibroids, among others. Antiestrogens, such as tamoxifen, are widely used for the treatment of breast cancer. Tamoxifen produces objective tumor shrinkage in advanced breast cancer, reduces the risk of relapse in women treated for invasive breast cancer, and prevents breast cancer in high risk women. Although, initially developed as an antiestrogen, tamoxifen can also prevent postmenopausal osteoporosis as well as reduce cholesterol, due to its estrogen-agonist effects. Its estrogen-agonist activity, however, can lead to significant side-effects such as endometrial cancer and thromboembolic phenomena. This has led to the concept of "ideal" selective estrogen receptor modulators (SERMs), drugs that would have the desired, tissue selective, estrogen-agonist or -antagonist effects. Raloxifene is a SERM which has the desirable mixed agonist/antagonist effects of tamoxifen but does not cause uterine stimulation. "Pure" antiestrogens may provide very potent estrogen-antagonist drugs, but are likely to be devoid of beneficial effects on bone and lipids. Future drug development efforts should focus on developing superior SERMs that have a greater efficacy against ER-positive tumors and do not cause hot flashes or thromboembolism, and explore combination strategies to simultaneously target hormone-dependent as well as hormone-independent breast cancer. PMID- 10665484 TI - Cerebral white matter lesions and cognitive function: the Rotterdam Scan Study. AB - Cerebral white matter lesions (WMLs) have been associated with cognitive dysfunction. Whether periventricular or subcortical WMLs relate differently to cognitive function is still uncertain. In addition, it is unclear whether WMLs are related to specific cognitive domains such as memory or psychomotor speed. We examined the relationship between periventricular and subcortical WMLs and cognitive functioning in 1,077 elderly subjects randomly sampled from the general population. Quantification of WMLs was assessed by means of an extensive rating scale on 1.5-T magnetic resonance imaging scans. Cognitive function was assessed by using multiple neuropsychological tests from which we constructed compound scores for psychomotor speed, memory performance, and global cognitive function. When analyzed separately, both periventricular and subcortical WMLs were related to all neuropsychological measures. When periventricular WMLs were analyzed conditional on subcortical WMLs and vice versa, the relationship between periventricular WMLs and global cognitive function remained unaltered whereas the relationship with subcortical WMLs disappeared. Subjects with most severe periventricular WMLs performed nearly 1 SD below average on tasks involving psychomotor speed, and more than 0.5 SD below average for global cognitive function. Tasks that involve speed of cognitive processes appear to be more affected by WMLs than memory tasks. PMID- 10665485 TI - Congenital muscular dystrophy with rigid spine syndrome: a clinical, pathological, radiological, and genetic study. AB - Rigid spine syndrome is a term first proposed by Dubowitz to describe a subset of patients affected by myopathy with early spinal contractures as a prominent feature. While spinal rigidity is a nonspecific feature, found in Emery-Dreifuss muscular dystrophy and in some congenital myopathies, it is also a prominent feature in a group of patients with merosin-positive congenital muscular dystrophy, where it is generally associated with stable or only slowly progressive weakness and early respiratory insufficiency. Recently, the first locus for congenital muscular dystrophy in association with rigid spine syndrome was mapped to chromosome 1p35-p36 in consanguineous Moroccan, Turkish, and Iranian families. We present here a detailed phenotypic description of the familial syndrome linked to this locus, describing 4 siblings (3 boys and 1 girl) of Northern European-American heritage who are the offspring of a nonconsanguineous marriage. All 4 siblings were affected by hypotonia and prominent neck weakness in infancy, early spinal rigidity, and early scoliosis. After initial improvement, muscle strength stabilizes or slowly declines, and skeletal deformities and respiratory insufficiency supervene. Muscle biopsy in an affected child at age 9 months revealed minimal, nonspecific myopathic changes, leading to a diagnosis of "minimal change myopathy." Muscle biopsy in his sibling, at the age of 14 years, revealed chronic and severe myopathic (dystrophic) changes, with normal staining for laminin-2 and for proteins of the dystrophin-glycoprotein complex. A possible explanation for these biopsy findings is that magnetic resonance imaging of the thighs reveals stereotyped selective muscle involvement, with the selectivity more pronounced early in the disease course followed by widespread muscular signal abnormalities in the late stages of the disease. In this family, linkage to the chromosome 1p rigid spine syndrome locus (RSMD1) is supported by maximum LOD scores for several markers of 1.81 at theta = 0, representing the maximum statistical power possible for this family. In combination with the previous report, this syndrome is linked to the RSMD1 locus with a summated maximum LOD score of 6.29, and analysis of recombination events in our family narrows the previously reported RSMD1 locus to 3 centiMorgans. PMID- 10665486 TI - The spectrum of mutations causing end-plate acetylcholinesterase deficiency. AB - The end-plate species of acetylcholinesterase (AChE) is an asymmetric enzyme consisting of a collagenic tail subunit composed of three collagenic strands (ColQ), each attached to a tetramer of the T isoform of the catalytic subunit (AChE(T)) via a proline-rich attachment domain. The principal function of the tail subunit is to anchor asymmetric AChE in the synaptic basal lamina. Human end plate AChE deficiency was recently shown to be caused by mutations in COLQ. We here report nine novel COLQ mutations in 7 patients with end-plate AChE deficiency. We examine the effects of the mutations on the assembly of asymmetric AChE by coexpressing each genetically engineered COLQ mutant with ACHE(T) in COS cells. We classify the newly recognized and previously reported COLQ mutations into four classes according to their position in ColQ and their effect on AChE expression. We find that missense mutations in the proline-rich attachment domain abrogate attachment of catalytic subunits, that truncation mutations in the ColQ collagen domain prevent the assembly of asymmetric AChE, that hydrophobic missense residues in the C-terminal domain prevent triple helical assembly of the ColQ collagen domain, and that other mutations in the C-terminal region produce asymmetric species of AChE that are likely insertion incompetent. PMID- 10665487 TI - Impaired reading in patients with right hemianopia. AB - A left occipital stroke may result in alexia for two reasons, which may coexist depending on the distribution of the lesion. A lesion of the left lateroventral prestriate cortex or its afferents impairs word recognition ("pure" alexia). If the left primary visual cortex or its afferents are destroyed, resulting in a complete right homonymous hemianopia, rightward saccades during text reading are disrupted ("hemianopic" alexia). By using functional imaging, we showed two separate but interdependent systems involved in reading. The first, subserving word recognition, involved the representation of foveal vision in the left and right primary visual cortex and the ventral prestriate cortex. The second system, responsible for the planning and execution of reading saccades, consisted of the representation of right parafoveal vision in the left visual cortex, the bilateral posterior parietal cortex (left > right), and the frontal eye fields (right > left). Disruption of this distributed neural system was demonstrated in patients with severe right homonymous hemianopia, commensurate with their inability to perform normal reading eye movements. Text reading, before processes involved in comprehension, requires the integration of perceptual and motor processes. We have demonstrated these distributed neural systems in normal readers and have shown how a right homonymous hemianopia disrupts the motor preparation of reading saccades during text reading. PMID- 10665488 TI - Oxidative phosphorylation defect in the brains of carriers of the tRNAleu(UUR) A3243G mutation in a MELAS pedigree. AB - MELAS is a mitochondrial encephalomyopathy characterized clinically by recurrent stroke-like episodes, seizures, sensorineural deafness, dementia, hypertrophic cardiomyopathy, and short stature. The majority of patients are heteroplasmic for a mutation (A3243G) in the tRNAleu(UUR) gene in mitochondrial DNA (mtDNA). In cells cultured in vitro, the mutation produces a severe mitochondrial translation defect only when the proportion of mutant mtDNAs exceeds 95% of total mtDNAs. However, most patients are symptomatic well below this threshold, a paradox that remains unexplained. We studied the relationship between the level of heteroplasmy for the mutant mtDNA and the clinical and biochemical abnormalities in a large pedigree that included 8 individuals carrying the A3243G mutation, 4 of whom were asymptomatic. Unexpectedly, we found that brain lactate, a sensitive indicator of oxidative phosphorylation dysfunction, was linearly related to the proportion of mutant mtDNAs in all individuals carrying the mutation, whether they were symptomatic or not. There was no evidence for threshold expression of the metabolic defect. These results suggest that marked tissue-specific differences may exist in the pathogenic expression of the A3243G mutation and explain why a neurological phenotype can be observed at relatively low levels of heteroplasmy. PMID- 10665489 TI - Synergistic neurotoxicity by human immunodeficiency virus proteins Tat and gp120: protection by memantine. AB - Human immunodeficiency virus type 1 (HIV-1) proteins Tat and gp120 have been implicated in the pathogenesis of dementia associated with HIV infection. Recently, we showed the presence of Tat protein in brains of patients with HIV-1 encephalitis as well as macaques with encephalitis caused by a chimeric strain of HIV and simian immunodeficiency virus, and that even transient exposure of cells to Tat leads to release of cytopathic cytokines. Now, we report the first demonstration of gp120 protein in brain of patients with HIV encephalitis. We tested the hypothesis that Tat and gp120 would act synergistically to potentiate each protein's neurotoxic effects and determined the extent to which pharmacological antagonists against processes implicated in HIV-1 neuropathogenesis could block HIV-1 protein-induced neurotoxicity. Subtoxic concentrations of Tat and gp120, when incubated together, caused neuronal cell death and prolonged increases in levels of intracellular calcium. A transient exposure of neurons to Tat and gp120 for seconds initiated neuronal cell death, but maximal levels of neuronal cell death were observed with exposures lasting 30 minutes. The neurotoxicity caused by Tat and gp120 applied in combination was blocked completely by memantine, partially by amiloride, and not at all by dipyridamole or vigabatrin. PMID- 10665490 TI - Prognostic value of proton magnetic resonance spectroscopic imaging for surgical outcome in patients with intractable temporal lobe epilepsy and bilateral hippocampal atrophy. AB - The objective of this study was to assess which features of temporal lobe proton magnetic resonance spectroscopic imaging (1H-MRSI) are associated with satisfactory surgical outcome in patients with intractable temporal lobe epilepsy and bilateral hippocampal atrophy. We studied 21 patients with intractable temporal lobe epilepsy and bilateral hippocampal atrophy defined by magnetic resonance imaging volumetric measurements who underwent surgical treatment. 1H MRSI was used to determine the relative resonance intensity ratio of the neuronal marker N-acetylaspartate to creatine + phosphocreatine (NAA/Cr) for mid and posterior temporal lobe regions of the left and right hemisphere, as well as an asymmetry index. Values lower than 2 SDs below the normal mean were considered abnormal. We used Engel's classification to assess surgical outcome with respect to seizure control. Eleven patients (52%) were in class I-II and 10 (48%) were in class III-IV. All 21 were operated on the side of maximal electroencephalographic (EEG) lateralization. Concordant lateralization of decreases in NAA/Cr to the side of surgery and normal NAA/Cr values in the contralateral posterior-temporal region were significantly associated with good surgical outcome: 11 (69%) of 16 patients with 1H-MRSI abnormalities concordant with EEG lateralization and none of the 5 patients with nonconcordant 1H-MRSI had a good outcome (class I-II); 10 (77%) of 13 patients with normal NAA/Cr contralateral to the EEG lateralization versus 1 (12.5%) of 8 of those with NAA/Cr reduction contralateral to EEG lateralization were in class I-II. Regression correlation analysis showed significant linear correlation between the midtemporal NAA/Cr relative asymmetry ratio and surgical outcome; the greater the asymmetry, the better the outcome. We conclude that discriminant 1H-MRSI features associated with favorable surgical outcome in patients with temporal lobe epilepsy and bilateral hippocampal atrophy were (1) concordant 1H-MRSI lateralization, (2) a greater side-to-side asymmetry of NAA/Cr, and (3) an absence of contralateral posterior NAA/Cr reduction. PMID- 10665491 TI - Expression of alpha-synuclein, parkin, and ubiquitin carboxy-terminal hydrolase L1 mRNA in human brain: genes associated with familial Parkinson's disease. AB - Mutations in the alpha-synuclein, parkin, and ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) genes have been linked to some cases of familial Parkinson's disease. To provide insight into how these genes may relate to each other and contribute to the pathology of Parkinson's disease, their expression was examined in normal human brain. Tissue sections from multiple regions of 11 normal human brains were hybridized with radiolabeled and digoxygenin-labeled cRNA probes for alpha-synuclein, parkin, and UCH-L1 mRNA. Expression of each of these three genes was predominantly neuronal. Alpha-synuclein and parkin mRNAs were expressed in a restricted number of brain regions, whereas UCH-L1 mRNA was more uniformly expressed throughout brain. The melanin-containing dopamine neurons of the substantia nigra had particularly robust expression. The expression patterns of alpha-synuclein and parkin mRNAs were similar, suggesting that these two proteins may be involved in common pathways contributing to the pathophysiology of Parkinson's disease. PMID- 10665492 TI - Molecular analysis of the CDR3 encoding region of the immunoglobulin heavy chain locus in cerebrospinal fluid cells as a diagnostic tool in lymphomatous meningitis. AB - The diagnosis of leptomeningeal B-cell lymphoma is based on the identification of malignant B cells in the cerebrospinal fluid (CSF). Frequently, cytology does not allow clear distinction between neoplastic lymphoid cells and reactively transformed mononuclear cells. Individual B-cell clones can be identified on the basis of DNA sequences that encode the highly diverse complementarity-determining region (CDR3) of the immunoglobulin heavy chain locus (IgH). We studied CSF samples from 5 patients with B-cell malignancies and cytological evidence of leptomeningeal involvement, using polymerase chain reaction (PCR)-based high resolution capillary electrophoresis and automated fluorescence analysis to detect PCR fragments. As controls, we assessed CSF specimens from 7 patients with inflammatory neurological diseases and three samples without pathological findings. In all patients with B-cell malignancies, a single PCR product was generated, indicating that CDR3-specific fragments were derived from monoclonal cell populations. CSF samples from patients with inflammatory diseases yielded multiple CDR3 amplicons, suggesting the presence of a polyclonal B-cell activation. No PCR product could be amplified in normal CSF samples. Automated fluorescence detection of CDR3 fragments is a highly sensitive and rapid method to distinguish neoplastic monoclonal and reactive polyclonal B-cell populations in the CSF. PMID- 10665493 TI - Joint position sense is impaired by Parkinson's disease. AB - The abilities of Parkinson's disease (PD) patients, taking routine medication, and of control subjects, to discriminate bilateral differences in the static angular positions of the two elbow joints were studied during passive (subject relaxed) and active (subject contracting to hold position) conditions. On each trial, one of the subject's elbows served as the reference joint (angle 60 degrees) and the other as the test joint (angular range, 54 degrees to 69 degrees, at 3 degree intervals). Subjects, with eyes closed, were required to discriminate the relative angles of the two elbows. In Experiments 1 (passive condition) and 2 (active condition), parkinsonians (n = 12) gave significantly fewer correct responses, pooled across sides, than did controls (n = 12), both in total scores across all angles and at individual test angles of 57 degrees and 63 degrees. In Experiment 3 (passive condition), derivation of conventional psychophysical variables indicated that both the difference limen (DL; threshold) and Weber ratio (WR; discriminatory sensitivity, independent of absolute stimulus values; same as DL/PSE) values of patients (n = 6) were significantly larger than those of controls (n = 6), in the absence of a significant difference between groups in the point of subjective equality (PSE). Our results provide clear evidence of a quantitative impairment of joint position sense in PD patients. PMID- 10665494 TI - Profile of endothelial and leukocyte activation in Fabry patients. AB - Fabry disease is an X-linked recessive disorder resulting in the deposition of globotriaosylceramide in numerous cell types including vascular endothelial cells. Because this disease is associated with vascular injury and a high recurrence rate of thrombotic events, measurements of factors regulating endothelium and leukocyte interaction may provide insight into the mechanisms leading to a prothrombotic state. Twenty-five patients with Fabry disease and 25 control subjects participated in the study. Plasma from all 25 Fabry patients and 15 of the 25 controls were studied for multiple endothelial factors. Leukocyte integrins were measured by flow cytometry in 21 Fabry patients and 10 controls. The concentrations of soluble intercellular adhesion molecule-1, vascular cell adhesion molecule-1, P-selectin, and plasminogen activator inhibitor were significantly higher and thrombomodulin was significantly lower in Fabry patients. Expression of the integrin CD11b on monocytes was also significantly higher in the Fabry patients. This study reveals measurable evidence for endothelium and leukocyte activation that is consistent with a prothrombotic state in Fabry patients compared with controls. Further investigations of these findings may help to understand the mechanism of stroke in Fabry disease and provide indicators (or markers) of efficacy of future therapeutic intervention. PMID- 10665495 TI - Heterogeneity of T-lymphocyte function in primary progressive multiple sclerosis: relation to magnetic resonance imaging lesion volume. AB - We found that in vitro migration and interferon-gamma production by lymphocytes derived from primary progressive multiple sclerosis patients preselected on the basis of a high T2-weighted lesion volume (>10 cm3) on magnetic resonance imaging, were increased compared with that in primary progressive multiple sclerosis patients with a low T2-weighted lesion volume (<3 cm3) and controls. Whether the heterogeneity of immune function within the primary progressive population will correlate with response to therapy remains to be established. PMID- 10665496 TI - No acute antimigraine efficacy of CP-122,288, a highly potent inhibitor of neurogenic inflammation: results of two randomized, double-blind, placebo controlled clinical trials. AB - CP-122,288 is a highly potent inhibitor of neurogenic plasma extravasation in animal models at doses without vasoconstrictor effect. We evaluated the acute antimigraine efficacy of intravenous and oral CP-122,288 in two double-blind studies. In a crossover design, patients randomly received 31.25 microg of CP 122,288 intravenously, placebo, or both. In the oral study, patients received placebo or one of four doses of CP-122,288 between 3.125 and 312.5 microg, using a novel "up and down" design for randomization. Both studies were stopped prematurely when target efficacy could not be achieved. Responder rates were 29% for CP-122,288 versus 30% for placebo (difference, -1%; 95% CI, -24-22%; intravenous study) and an overall rate of 25% for CP-122,288 versus 0% for placebo (difference, 25%; 95% CI; 10-40%; oral study). CP-122,288 was not clinically effective at doses and plasma concentrations in excess of those required to inhibit neurogenic plasma extravasation in animals. Neurogenic plasma extravasation is unlikely to play a crucial role in the pathophysiology of migraine headache. PMID- 10665497 TI - Significant association between the tau gene A0/A0 genotype and Parkinson's disease. AB - A significant association between the tau gene A0/A0 genotype and progressive supranuclear palsy has been reported recently. To determine if the presence of a tau polymorphism could constitute a risk factor for the development of sporadic and familial Parkinson's disease, a dinucleotide repeat marker at intron 11 was genotyped in 152 patients with PD, 52 patients with Alzheimer's disease, and 150 healthy controls. We detected a significant difference in A0 allelic frequency in the Parkinson's disease group (79.27%) compared with the control group (71%) and the Alzheimer's disease group (73.07%). Individuals homozygous for the A0 allele were also detected significantly more frequently in the Parkinson's disease group (63.8%) compared with the control group (52.66%) and the Alzheimer's disease group (48.07%). These results suggest a possible involvement of the tau gene in the pathogenesis of some cases of Parkinson's disease. PMID- 10665498 TI - Major depression is a risk factor for seizures in older adults. AB - We tested the hypothesis that major depression meeting DSM-III-R criteria or medical therapies for depression increase the risk for unprovoked seizures. Major depression was associated with a sixfold increased risk for unprovoked seizures (95% CI, 1.56-22). The risk remained increased even when controlling for age, sex, length of medical follow-up, and medical therapies for depression. In the absence of known prior neurological insult, major depression is associated with an increased risk for unprovoked seizures. PMID- 10665499 TI - Novel Leu723Pro amyloid precursor protein mutation increases amyloid beta42(43) peptide levels and induces apoptosis. AB - A novel missense mutation, Leu723Pro, in the amyloid precursor protein (APP) gene was discovered in an early-onset Alzheimer's disease family. Expression of L723P mutant APP complementary DNA in CHO cells resulted in a 1.4- to 1.9-fold increased production of the 42(43)-amino acid length amyloid beta peptide compared with the wild-type sequence and was capable of causing apoptosis. The mutation is predicted to alter the luminal transmembrane length and helical arrangement of the APP molecule and thus affect the gamma-secretase cleavage site. PMID- 10665500 TI - Prenatal testing for late infantile neuronal ceroid lipofuscinosis. AB - Classic late infantile neuronal ceroid lipofuscinosis (LINCL) is a neurodegenerative disease in which autofluorescent "curvilinear" storage bodies accumulate in tissues from affected patients. Recently, the LINCL gene (CLN2) has been found to code for a pepstatin-insensitive lysosomal protease whose activity is deficient in LINCL specimens. We report the first 2 cases of successful prenatal testing for LINCL by using DNA and enzyme-based methods on amniocytes, and describe a new private mutation in one of the families analyzed. These approaches allow definitive prenatal diagnosis and represent a significant improvement over previous pathological methods. PMID- 10665501 TI - Creutzfeldt-Jakob disease profile in patients homozygous for the PRNP E200K mutation. AB - We identified 70 Creutzfeldt-Jakob disease patients with the previously described E200K mutation in the prion protein gene. The purpose of this study was to define the clinical features of E200K homozygous patients (n = 5), compared with heterozygotes. We found a statistically significant younger age at disease onset for the homozygous patients, although the average age at onset in this group was still in midlife. Disease features were not statistically different in the two groups. Possible explanations are discussed. PMID- 10665502 TI - Vitamin E deficiency due to chylomicron retention disease in Marinesco-Sjogren syndrome. AB - We report on 2 brothers (aged 19 and 12 years) with Marinesco-Sjogren syndrome who also had very low serum vitamin E concentrations with an absence of postprandial chylomicrons. The molecular study ruled out ataxia with isolated vitamin E deficiency, abetalipoproteinemia, and hypobetalipoproteinemia. The electron microscopy of the intestinal mucosa was consistent with a chylomicron retention disease. We speculate that both chylomicron retention disease and Marinesco-Sjogren syndrome are related to defects in a gene crucial for the assembly or secretion of the chylomicron particles, leading to very low serum levels of vitamin E. PMID- 10665503 TI - Genetic and neuroradiological heterogeneity of double cortex syndrome. AB - Mutations in the X-linked doublecortin gene appear in many sporadic cases of double cortex (DC; also known as subcortical band heterotopia), a neuronal migration disorder causing epilepsy and mental retardation. The purpose of this study was to examine why a significant percentage of sporadic DC patients had been found not to harbor doublecortin mutations and to determine whether clinical features or magnetic resonance imaging scan appearance could distinguish between patients with and without doublecortin mutations. Magnetic resonance imaging scan analysis differentiated patients into the following four groups: anterior biased/global DC with doublecortin mutation (16 of 30; 53%), anterior biased/global DC without mutation (8 of 30; 27%), posterior biased DC without mutation (3 of 30; 10%), and limited/unilateral DC without mutation (3 of 30; 10%). The presence of these atypical phenotypes suggests that other genetic loci or mosaicism at the doublecortin locus may be responsible for this diversity of DC cases. PMID- 10665504 TI - Increased numbers of CCR5+ interferon-gamma- and tumor necrosis factor-alpha secreting T lymphocytes in multiple sclerosis patients. AB - To determine the frequency of in vivo activated T(H)1 lymphocytes, T-cell subsets of 9 multiple sclerosis patients with active disease and 17 healthy controls were analyzed by immunostaining for CCR5, CD26, and their expression of interleukin-2, interferon-gamma, and tumor necrosis factor-alpha. The numbers of CCR5+ interferon-gamma- and tumor necrosis factor-alpha-producing T cells were significantly increased in the peripheral blood of multiple sclerosis patients. CCR5 expression may be a useful marker to identify effector cells in multiple sclerosis and could be used as a tool for monitoring disease activity. PMID- 10665505 TI - No cytomegalovirus DNA in sera from patients with anti-MAG/SGPG antibody associated neuropathy. PMID- 10665506 TI - The Pocket Monsters episode. PMID- 10665507 TI - Seasonal fluctuations of gadolinium-enhancing magnetic resonance imaging lesions in multiple sclerosis. PMID- 10665508 TI - Intracellular adhesion molecule-1 polymorphisms and genetic susceptibility to multiple sclerosis: additional data and meta-analysis. PMID- 10665509 TI - Subtle brain abnormalities in children with sickle cell disease: relationship to blood hematocrit. PMID- 10665510 TI - Effect of vowel environment on consonant duration: an extension of normative data to adult contextual speech. AB - In this study, we investigated the effect of vowel environment on consonant duration in contextual speech produced by adults. Previous studies, such as Schwartz's published in 1969 and DiSimoni's in 1974, of vowel influence on consonant duration have supported the notion of anticipatory scanning in which final vowel targets influence the duration of preceding fricative consonants. These studies were based on repetitions of nonsense syllables by children and adults, but no research has been reported that extends these data to contextual speech or examines speaker gender differences. Forty adult normal speakers (20 women and 20 men) recorded palatal and alveolar fricatives produced in four vowel environments in words embedded in contextual sentences. Results indicated significant effects of vowel context on consonant duration in contextual speech and revealed anticipatory scanning effects that are similar to those seen with nonsense syllables in previous studies. These normative data can form the basis for comparison of the effects of temporal alterations produced by speaking conditions such as simultaneous communication. PMID- 10665511 TI - Academic outcomes in children with histories of speech sound disorders. AB - Tests of phonology, semantics, and syntax were administered to 52 preschool children (19 girls and 33 boys, age 4-6 years) with moderate to severe speech sound disorders. The children's performance on these tests was used to predict language, reading, and spelling abilities at school age (age 8-11 years). Language impairment at school age was related to poor performance on preschool tests of syntax and nonsense word repetition, while reading impairment was predicted by poor performance in all preschool test domains (phonology, semantics, and syntax). In contrast, spelling impairment was predicted by deficits in preschool tests of phonological processing as measured by the Word Discrimination subtest of the Test of Language Development - Primary 2. Family history for speech and language disorders did not predict language, reading, or spelling impairment at school age. However, family history for reading disorders was a good predictor of school-age spelling difficulties. PMID- 10665512 TI - Cultural analysis of communication behaviors among juveniles in a correctional facility. AB - This study addressed communication behaviors of female juvenile delinquents in a correctional facility. Qualitative methodology was used to study 78 participants ranging in age from 13.1 to 18.9 (years; months), over a five-month period. Data collection consisted of observations, participant observation, interviews, and a review of documents. Additionally, participants were tested on the Clinical Evaluation of Language Fundamentals-3. Listening and following rules, utterance types, topics of conversion, politeness, and conversational management emerged as themes. Findings indicated that as many as 22% of participants were potential candidates for language services. Implications for speech-language pathologists (SLPs) providing communication services will be provided. PMID- 10665513 TI - Effects of speech therapy and pharmacologic and surgical treatments on voice and speech in Parkinson's disease: a review of the literature. AB - The purpose of this review was to examine the different treatment approaches for persons with Parkinson's Disease (PD) and to examine the effects of these treatments on speech. Treatment methods reviewed include speech therapy, pharmacological, and surgical. Research from the 1950s through the 1970s had not demonstrated significant improvements following speech therapy. Recent research has shown that speech therapy (when persons with PD are optimally medicated) has proven to be the most efficacious therapeutic method for improving voice and speech function. Pharmacological methods of treatment in isolation do not appear to significantly improve voice and speech function in PD across research studies. Surgical treatment methods including pallidotomy and deep brain stimulation may be significant treatment options which improve voice and speech function in some persons with PD. Possible explanations for the differential responses to treatment are discussed. Future studies should investigate the effects of combined treatment approaches. Perhaps the combination of pharmacological, surgical and speech treatment will prove superior to treatments combining pharmacological and surgical or pharmacological and speech therapy in improving the communication abilities of persons with PD. PMID- 10665514 TI - Preclinical overview of brinzolamide. AB - The development of topically active carbonic anhydrase inhibitors (CAIs) is a significant recent achievement in glaucoma medical treatment. Brinzolamide, the newest topical CAI, exhibits selectivity, high affinity, and potent inhibitory activity for the carbonic anhydrase type II isozyme (CA-II), which is involved in aqueous humor secretion. These characteristics, along with good ocular bioavailability, make brinzolamide maximally effective in lowering intraocular pressure (IOP) by locally inhibiting CA-II in the ciliary processes and suppressing aqueous humor secretion. Notable among its attributes as a safe and efficacious glaucoma drug is brinzolamide's superior ocular comfort profile because of its optimized suspension formulation at physiologic pH. The degree of tolerability in the eye is considered an important determinant of a patient's willingness to comply with the dosing regimen for a long-term glaucoma medication. Results from the preclinical pharmacologic evaluation of brinzolamide indicated that it acts specifically to inhibit CA without significant other pharmacologic actions that could introduce undesired side effects. Moreover, the typical side effects associated with systemically administered CAIs are expected to occur at a lower incidence or not occur at all with brinzolamide, as its therapeutic dose and low systemic absorption do not produce a problematic level of systemic CA inhibition. Brinzolamide's long tissue half-life in the eye, particularly in the iris-ciliary body, favors a prolonged duration of IOP lowering. This was substantiated in clinical trials, which showed that twice daily brinzolamide provides as significant an IOP reduction as three-times-daily brinzolamide or dorzolamide in a relatively high percentage of patients. Brinzolamide has been shown by the laser Doppler flowmetry technique to improve blood flow to the optic nerve head in pigmented rabbits after topical administration, without producing an increase of blood pCO2, indicating a potential for a local vasodilatory effect involving the optic nerve head circulation. The mean concentration of brinzolamide found in the retina of pigmented rabbits (0.338 microg equivalents/g) after a single dose of 14C brinzolamide is sufficient to inhibit CA-II. These data suggest that topical brinzolamide could improve the blood flow in the optic nerve head in humans should it inhibit carbonic anhydrase in that vascular bed. Brinzolamide is a new topically active CAI that is safe and efficacious for reducing intraocular pressure. It offers the convenience of topical dose administration and greater freedom from side effects related to the inhibition of CA seen with the systemic administration of CAIs. Its formulation has been optimized to provide greater comfort upon instillation, and this can result in a higher compliance rate by the patient. Results of studies in animals show that brinzolamide has promise for increasing blood flow to the optic nerve head; however, this requires further assessment in the clinic. Brinzolamide represents a significant technical achievement and an important addition to the medical treatment of glaucoma as both a primary and an adjunctive drug. PMID- 10665515 TI - Increased optic nerve head blood flow after 1 week of twice daily topical brinzolamide treatment in Dutch-belted rabbits. AB - OBJECTIVES: Using a three-way crossover study design, we compared the effects of brinzolamide 2%, dorzolamide 2%, and placebo (vehicle) on microvascular optic nerve head (ONH) blood flow, intraocular pressure (IOP), blood pressure, heart rate, and acid-base balance in nine acepromazine-tranquilized Dutch-belted rabbits. METHODS: Baseline measurements were taken before treatment and after drug-free washout periods of 7-14 days. Microvascular ONH blood flow was measured with a fundus camera-based laser Doppler flowmeter (LDF). Intraocular pressure was measured with a Tono-Pen XL. One drop of brinzolamide, dorzolamide, or vehicle was administered twice daily (9 A.M. and 5 P.M.) in right eyes only for 7 days. Experimental measurements were made 90 minutes after the 9 A.M. topical dose was administered on day 8. RESULTS: ONH blood flow was significantly increased (P< or =0.05) in carbonic anhydrase inhibitor (CAI)-treated rabbits, as compared with vehicle-treated controls. The percent increase from baseline was 11.2+/-1.8% in brinzolamide-treated animals and 8.4+/-4.3% in dorzolamide-treated animals. Compared with controls, IOP in the brinzolamide- and dorzolamide-treated groups was significantly decreased (P< or =0.05). The changes in ONH blood flow and IOP were not significantly different between the CAI treatment groups. Small but significant changes in systemic blood gas tensions and pH were present in both CAI treatment groups, as compared with the vehicle group. Systemic blood pressure and heart rate were not significantly changed. CONCLUSIONS: Topical ocular CAI treatment for 1 week with either brinzolamide or dorzolamide significantly reduced IOP and significantly increased ONH blood flow in tranquilized Dutch-belted rabbits, while eliciting minimal systemic acid-base balance disturbances. PMID- 10665516 TI - Ocular comfort of brinzolamide 1.0% ophthalmic suspension compared with dorzolamide 2.0% ophthalmic solution: results from two multicenter comfort studies. Brinzolamide Comfort Study Group. AB - Two independent, prospective, multicenter, double-masked, parallel group trials were conducted to compare the ocular comfort of brinzolamide 1.0% administered three times daily (t.i.d.) with t.i.d.-dosed dorzolamide 2.0% in patients with primary open-angle glaucoma or ocular hypertension. Patients were randomized to one of two treatment groups, receiving either brinzolamide 1.0% t.i.d. or dorzolamide 2.0% t.i.d. for 1 week. On the last day of dosing, patients received one drop of masked medication in both eyes, and ocular discomfort (burning or stinging) was evaluated by means of a 4-unit ocular discomfort scale. The incidence and extent of ocular discomfort across both treatment groups were analyzed. The results from both studies were confirmatory and demonstrated that the ocular discomfort score for brinzolamide 1.0% was 1.3 units lower than the score for dorzolamide 2.0%, which was both statistically significant and clinically relevant. In addition, a statistically significantly greater percentage of patients reported no ocular discomfort with brinzolamide 1.0% compared with dorzolamide. A greater percentage of patients receiving dorzolamide 2.0% also reported mild, moderate, severe, and very severe ocular discomfort compared with those treated with brinzolamide 1.0%. The most frequent ocular adverse event reported in the brinzolamide group was transient blurred vision, which ranged from 20% to 25%. Overall, adverse events associated with brinzolamide 1.0% and dorzolamide 2.0% were nonserious, were usually mild, and resolved without treatment. The findings of each study independently demonstrated that brinzolamide 1.0% was significantly more comfortable than dorzolamide 2.0% when instilled in the eye. PMID- 10665517 TI - Dose-response evaluation of the ocular hypotensive effect of brinzolamide ophthalmic suspension (Azopt). Brinzolamide Dose-Response Study Group. AB - Brinzolamide is a novel carbonic anhydrase inhibitor that elicits an ocular hypotensive effect when instilled topically. A multicenter, double-masked, placebo-controlled, parallel trial was conducted to evaluate the optimal intraocular pressure (IOP)-lowering concentration and ocular tolerability of topically administered brinzolamide (0.3%, 1%, 2%, and 3%) in patients with primary, open-angle glaucoma or ocular hypertension. After a washout phase, patients were administered brinzolamide or placebo twice daily for 2 weeks. The IOP was measured on days 8 and 15 at 8:00 A.M., and then 2, 4, 8, and 12 hours after dosing, and these measurements were compared with IOP values obtained at the corresponding times during an off-therapy diurnal baseline. All concentrations of brinzolamide produced significantly greater (P<0.005) mean percent IOP reductions and mean IOP reductions compared with placebo. Mean percent IOP changes (mean IOP changes) from baseline for brinzolamide 0.3%, 1%, 2%, and 3% were -11.3% (-3.0 mm Hg), -16.1% (-4.3 mm Hg), -16.1% (-4.4 mm Hg), and -15.4% (-4.2 mm Hg), respectively, when pooled over visit and visit time. Comparisons between concentrations demonstrated that the mean percent IOP reduction for brinzolamide 1.0% was significantly greater than that for the 0.3% concentration (P<0.03), with no difference in efficacy between the 1%, 2%, and 3% concentrations. The incidence of adverse events was dose-dependent, and those related to therapy were usually mild and resolved without treatment. Blurred vision, ocular discomfort, and abnormal taste were the most frequently reported adverse events. Based on these findings, the optimal IOP-lowering concentration of brinzolamide was 1%. When administered twice daily, brinzolamide 1% was well tolerated by patients with primary open-angle glaucoma or ocular hypertension. PMID- 10665518 TI - The efficacy and safety of brinzolamide 1% ophthalmic suspension (Azopt) as a primary therapy in patients with open-angle glaucoma or ocular hypertension. Brinzolamide Primary Therapy Study Group. AB - A randomized, multicenter, double-masked, prospective, parallel study was designed to establish the intraocular pressure (IOP)-lowering efficacy, safety, and tolerability of brinzolamide 1.0% (Azopt) as a primary therapy compared with dorzolamide 2.0% (Trusopt) and placebo in patients diagnosed with open-angle glaucoma (with or without a pseudoexfoliative or a pigmentary dispersion component) or ocular hypertension. Brinzolamide 1.0%, dosed two times (b.i.d.) and three times (t.i.d.) a day, dorzolamide 2.0% (t.i.d.), and placebo (t.i.d) were administered to patients during a 3-month treatment period. Diurnally corrected IOP reduction from baseline, including peak and trough times, was the primary end point. Sample sizes were chosen to establish statistical equivalence between treatments. Mean IOP changes observed on treatment were as follows: -3.4 mm Hg (-13.2%) to -4.1 mm Hg (-16.7%) with brinzolamide 1.0% b.i.d.; -4.1 mm Hg ( 16.6%) to -4.8 mm Hg (-19.1%) with brinzolamide 1% t.i.d.; and -4.3 mm Hg ( 16.9%) to -4.9 mm Hg (-20.1%) with dorzolamide 2.0%. IOP reductions after administration of brinzolamide 1.0% b.i.d. and t.i.d. were equivalent to each other and also clinically and statistically equivalent to those with dorzolamide 2.0% t.i.d. The incidence of ocular discomfort (burning and stinging) upon instillation was significantly higher for dorzolamide (10.7%) than brinzolamide (b.i.d. or t.i.d., 3.0% each). The most frequent non-ocular event reported was taste perversion, which was less (3.7%) with brinzolamide 1.0% b.i.d., but brinzolamide t.i.d. was similar to dorzolamide t.i.d. (6.8% vs. 5.3%). Brinzolamide 1.0% b.i.d., brinzolamide 1.0% t.i.d., and dorzolamide 2.0% t.i.d. equaled each other in IOP-lowering efficacy, and brinzolamide was significantly more comfortable than dorzolamide upon instillation. PMID- 10665519 TI - Adjunctive therapy with brinzolamide 1% ophthalmic suspension (Azopt) in patients with open-angle glaucoma or ocular hypertension maintained on timolol therapy. AB - This prospective, multicenter, double-masked, placebo-controlled study evaluated the safety and efficacy of brinzolamide 1% ophthalmic suspension (Azopt) when used adjunctively with open-label timolol maleate 0.5% (Timoptic). One-hundred thirty-two patients requiring an adjunctive therapy to timolol 0.5% for the treatment of open-angle glaucoma or ocular hypertension were randomized to receive brinzolamide or placebo three times daily (t.i.d.) in addition to timolol 0.5% twice daily (b.i.d.) for 3 months. Qualifying intraocular pressure (IOP) on timolol 0.5% b.i.d. was 24-36 mm Hg in at least one eye at 8:00 A.M. and 21-36 mm Hg at 10:00 A.M., with no greater than a 5-mm Hg difference between eyes, during two eligibility visits separated by at least 7 days. Treatments were compared using a repeated-measures analysis of variance. Adjunctive therapy with brinzolamide resulted in clinically and statistically significant reductions in IOP from the timolol baseline at all visits. IOP changes from a diurnal baseline ranged from -3.3 mm Hg to -4.1 mm Hg for brinzolamide (N = 53) compared with -0.9 mm Hg to -2.5 mm Hg for placebo (N = 55). Abnormal taste (7.7%) and transient blurred vision (6.2%) were the most frequently reported adverse events. No clinically significant differences in the incidence or severity of ocular signs, visual acuity, cup/disk ratio, or parameters studied on dilated fundus examination were observed between treatment groups. Brinzolamide 1% t.i.d., used adjunctively with timolol 0.5% b.i.d., is safe and well tolerated, and produces clinically and statistically significant additional IOP reductions. PMID- 10665520 TI - Mitochondrial events in the life and death of animal cells: a brief overview. AB - Traditionally, mitochondria have been viewed as the "powerhouse" of the cell, i.e., the site of the oxidative phosphorylation machinery involved in ATP production. Consequently, much of the research conducted on mitochondria over the past 4 decades has focused on elucidating both those molecular events involved in ATP synthesis by oxidative phosphorylation and those involved in the biogenesis of the oxidative phosphorylation machinery. While monumental achievements have been made, and continue to be made, in the study of these remarkable but extremely complex processes essential for the life of most animal cells, it has been only in recent years that a large body of biological and biomedical scientists have come to recognize that mitochondria participate in other important processes. Two of these are cell death and aging which, not surprisingly, are related processes both involving, in part, the oxidative phosphorylation machinery. This new awareness has sparked a new and growing area of mitochondrial research, that has become of great interest to a wide variety of scientists ranging from those involved in elucidating the role of mitochondria in cell death and aging to those interested in either suppressing or facilitating these processes as it relates to identifying new therapies or drugs for human disease. It is the purpose of this brief introductory review to provide an overview of those mitochondrial events involved in the life and death of animal cells and to indicate how these events might relate to the human aging process. Much more is known, much remains controversial, and even more remains to be learned as indicated in the excellent set of minireviews that follow. PMID- 10665521 TI - Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death. AB - Mitochondria are frequently the target of injury after stresses leading to necrotic and apoptotic cell death. Inhibition of oxidative phosphorylation progresses to uncoupling when opening of a high conductance permeability transition (PT) pore in the mitochondrial inner membrane abruptly increases the permeability of the mitochondrial inner membrane to solutes of molecular mass up to 1500 Da. Cyclosporin A (CsA) blocks this mitochondrial permeability transition (MPT) and prevents necrotic cell death from oxidative stress, Ca2+ ionophore toxicity, Reye-related drug toxicity, pH-dependent ischemia/reperfusion injury, and other models of cell injury. Confocal fluorescence microscopy directly visualizes onset of the MPT from the movement of green-fluorescing calcein into mitochondria and the simultaneous release from mitochondria of red-fluorescing tetramethylrhodamine methylester, a membrane potential-indicating fluorophore. In oxidative stress to hepatocytes induced by tert-butylhydroperoxide, NAD(P)H oxidation, increased mitochondrial Ca2+, and mitochondrial generation of reactive oxygen species precede and contribute to onset of the MPT. Confocal microscopy also shows directly that the MPT is a critical event in apoptosis of hepatocytes induced by tumor necrosis factor-alpha. Progression to necrotic and apoptotic cell killing depends, at least in part, on the effect the MPT has on cellular ATP levels. If ATP levels fall profoundly, necrotic killing ensues. If ATP levels are at least partially maintained, apoptosis follows the MPT. Cellular features of both apoptosis and necrosis frequently occur together after death signals and toxic stresses. A new term, necrapoptosis, describes such death processes that begin with a common stress or death signal, progress by shared pathways, but culminate in either cell lysis (necrosis) or programmed cellular resorption (apoptosis) depending on modifying factors such as ATP. PMID- 10665522 TI - Mitochondria at the crossroad of apoptotic cell death. AB - In the past few years, it has become widely appreciated that apoptotic cell death generally involves activation of a family of proteases, the caspases, which undermine the integrity of the cell by cleavage of critical intracellular substrates. Caspases, which are synthesized as inactive zymogens, are themselves caspase substrates and this cleavage leads to their activation. Hence, the potential exists for cascades of caspases leading to cell death. However, it has been recently recognized that another, perhaps more prominent route to caspase activation, involves the mitochondria. Upon receipt of apoptotic stimuli, either externally or internally generated, cells initiate signaling pathways which converge upon the mitochondria to promote release of cytochrome C to the cytoplasm; cytochrome c, thus released, acts as a potent cofactor in caspase activation. Even cell surface "death receptors" such as Fas, which can trigger direct caspase activation (and potentially a caspase cascade), appear to utilize mitochondria as part of an amplification mechanism; it has been recently demonstrated that activated caspases can cleave key substrates to trigger mitochondrial release of cytochrome c, thereby inducing further caspase activation and amplifying the apoptotic signal. Therefore, mitochondria play a central role in apoptotic cell death, serving as a repository for cytochrome c. PMID- 10665523 TI - Mitochondrial redox signaling during apoptosis. AB - The regulatory role of cellular redox state during apoptosis is still controversial. Early redox signaling can transduce divergent upstream signals to mitochondria and initiate apoptosis. On the other hand, release of mitochondrial cytochrome c triggers generation of reactive oxygen species (ROS) and renders apoptotic cells much more oxidized. Although the sequential caspase activation does not have apparent redox-sensitive components, redox signaling provides a separate pathway that is parallel with the caspase cascade. The function of the apoptosis-associated redox change is uncertain. It could provide positive feedback mechanisms, such as activating mitochondrial permeability transition and apoptosis signaling kinase (ASK-1). Since apoptotic cells are designated to be quickly eliminated, the dramatic cellular oxidation could be involved in the final degradation of apoptotic bodies and even the termination of the proteolytic activity after phagocytosis. PMID- 10665524 TI - Progress on the mitochondrial permeability transition pore: regulation by complex I and ubiquinone analogs. AB - This review summarizes recent progress on the regulation of the mitochondrial permeability transition pore, an inner membrane channel that may play a role in cell death. We briefly cover its key control points as emerged over the last few years from studies on isolated mitochondria; and describe in some detail our recent results indicating that the pore is modulated by the respiratory chain complex I and can be specifically blocked by selected ubiquinone analogs. We discuss the potential relevance of these findings for the structural definition of the permeability transition pore and illustrate the pharmacological perspectives they offer in diseases where mitochondrial dysfunction is suspected to play a key role. PMID- 10665525 TI - Mitochondrial oxygen radical generation and leak: sites of production in states 4 and 3, organ specificity, and relation to aging and longevity. AB - Studies in heart and nonsynaptic brain mitochondria from two mammals and three birds show that complex I generates oxygen radicals in heart and nonsynaptic brain mitochondria in States 4 and 3, whereas complex III does it only in heart mitochondria and only in State 4. The increase in oxygen consumption during the State 4 to 3 transition is not accompanied by a proportional increase in oxygen radical generation. This will protect mitochondria and tissues during bursts of activity. Comparisons between young and old rodents do not show a consistent pattern of variation in mitochondrial oxygen radical production during aging. However, all the interspecies comparisons performed to date between different mammals, and between mammals and birds, agree that animals with high maximum longevities have low rates of mitochondrial oxygen radical production, irrespective of the value of their basal specific metabolic rate. The sites and mechanisms allowing this, the recently described low degree of membrane fatty acid unsaturation of longevous animals, and their relation to longevity and aging are discussed. PMID- 10665526 TI - Cooperation of a "reactive oxygen cycle" with the Q cycle and the proton cycle in the respiratory chain--superoxide generating and cycling mechanisms in mitochondria. AB - Based on our recent findings concerning the generating, partitioning, targeting, and functioning of superoxide in mitochondria, a hypothetical model involving a "reactive oxygen cycle" in the respiratory chain has been proposed (Liu and Huang, 1991, 1996; Liu et al., 1996; Liu, 1997, 1998) This model emphasizes that during State 4 respiration, an interaction between an electron leak (a branch of electron transfer directly from the respiratory chain to form O2*-, but not H2O) and a proton leak (a branch pathway which utilizes delta muH+ to produce heat, but not ATP) may take place in cooperation with the Q and proton cycles in mitochondria through the consumption of H+ by O2*- anions to form a protonated perhydroxyl radical, HO2, which is directly permeable across the inner mitochondrial membrane and induces proton leakage and a decrease of delta muH+. O2*- generation in the mitochondrial respiratory chain and its cycling across the inner membrane may have the role of an endogenous protonophore in regulating and partitioning energy transduction and heat production, as well as in pathogenesis of mitochondrial diseases, aging, and apoptosis. The present article summarizes the supporting experimental evidence obtained in this laboratory and presents a brief description of the theoretical basis of this model. PMID- 10665527 TI - Mitochondrial genome mutation in cell death and aging. AB - This article reviews the concept, molecular genetics, and pathology of cell death and aging in relation to mitochondrial genome mutation. Accumulating evidence emphasizes the role of genetic factors in the development of naturally occurring cell death and aging. The ATP required for a cell's biological activity is almost exclusively produced by mitochondria. Each mitochondrion possesses its own DNA (mtDNA) that codes essential subunits of the mitochondrial energy-transducing system. Recent studies confirm that mtDNA is unexpectedly fragile to hydroxyl radical damage, hence to the oxygen stress. Cellular mtDNA easily fragments into over a hundred-types of deleted mtDNA during the life of an individual. Cumulative accumulation of these oxygen damages and deletions in mtDNA results in a defective energy transducing system and in bioenergetic crisis. The crisis leads cells to the collapse of mitochondrial trans-membrane potential, to the release of the apoptotic protease activating factors into cytosol, to uncontrolled cell death, to tissue degeneration and atrophy, and to aging. The total base sequencing of mtDNA among individuals revealed that germ-line point mutations transmitted from ancestors accelerate the somatic oxygen damages and mutations in mtDNA leading to phenotypic expression of premature aging and degenerative diseases. A practical survey of point mutations will be useful for genetic diagnosis in predicting the life-span of an individual. PMID- 10665528 TI - Mitochondrial DNA repair pathways. AB - It has long been held that there is no DNA repair in mitochondria. Early observations suggested that the reason for the observed accumulation of DNA damage in mitochondrial DNA is that DNA lesions are not removed. This is in contrast to the very efficient repair that is seen in the nuclear DNA. Mitochondrial DNA does not code for any DNA repair proteins, but it has been observed that a number of repair factors can be found in mitochondrial extracts. Most of these participate in the base excision DNA repair pathway which is responsible for the removal of simple lesions in DNA. Recent work has shown that there is efficient base excision repair in mammalian mitochondria and there are also indications of the presence of more complex repair processes. Thus, an active field of mitochondrial DNA repair is emerging. An understanding of the DNA repair processes in mammalian mitochondria is an important current challenge and it is likely to lead to clarification of the etiology of the common mutations and deletions that are found in mitochondria, and which are thought to cause various human disorders and to play a role in the aging phenotype. PMID- 10665529 TI - The stability of porcine rotavirus in feces. AB - Rotaviruses are known as major causal agents of diarrhea in humans and animals. They affect young animals in intensive rearing and cause great economic losses. This study evaluated the infectivity of porcine rotavirus maintained for 32 months at approximately 10 degrees C in the original stool specimens. Thirty stool specimens of 1-4-week-old piglets from breeding farms located in the southwest of the State of Parana were selected for this study. They were randomly chosen from stool samples positive for rotavirus RNA by polyacrylamide gel electrophoresis (PAGE) at the time of collection. The thirty stool samples maintained for 32 months were re-tested by PAGE and 11 out of 30 were still positive showing physical integrity of the eleven segments of viral RNA. In order to demonstrate the maintenance of viral infectivity processed fecal homogenates were inoculated in MA-104 cell cultures. After an average of three blind passages 5 out of 11 samples demonstrated cytopathic effect similar to that of a simian rotavirus (SA-11) used as positive control. To confirm these findings an immunofluorescence test was performed and typical cytoplasmatic granular fluorescence was observed. Electron microscopy of stool samples showed that most of the virus particles were single-shelled and some were found to be in advanced state of degradation. The viral nucleic acid extracted from six fecal specimens out of those that showed physical integrity of rotavirus RNA by PAGE were also amplified when submitted to RT-PCR demonstrating stability of viral RNA. We therefore concluded that porcine rotavirus infectivity is maintained for a long period of time in stool specimens at low temperature. PMID- 10665530 TI - Effects of porcine reproductive and respiratory syndrome virus (isolate tw91) on porcine alveolar macrophages in vitro. AB - To verify the role of porcine reproductive and respiratory syndrome virus (PRRSV) infection on pulmonary defense mechanisms, alterations in the viability, morphology, and various functions of porcine alveolar macrophages (AMs) were evaluated in vitro for 2-72 h after exposure to a Taiwan isolate, tw91, at a multiplicity of infection (MOI) of 0.1. A low but constant rate of infection, around 5%, was seen in AMs from the PRRSV-infected group throughout the study. When compared with a mock-infected group, AMs from the PRRSV-infected group had a significantly lower viability at 18-72 h post-infection (HPI) as determined by trypan blue dye exclusion. Also during this time period, the cells showed morphological changes, including rounding, bleb formation, and rupture. The phagocytic and microbicidal capacity of AMs against Candida albicans was significantly inhibited after 6 HPI. Although the total amount of superoxide anion (O2-) and hydrogen peroxide (H2O2) produced by the AMs was reduced after 18 and 12 HPI, respectively, the amount of production was enhanced in both reactive oxygen species on a per viable cell basis after 12 HPI. In contrast, the level of bioactive tumor necrosis factor alpha (TNF-alpha) secretion, either total or on a per viable cell basis, was markedly reduced soon after PRRSV infection, up to 36 HPI, followed by a rebound thereafter. Prostaglandin E2 (PGE2) production was enhanced, both in total and on a per viable cell basis, in the first 6 h of infection, especially at 2 HPI. However, it became lower than that of the control after 36 HPI. The results indicated that PRRSV infection could cause, directly and/or indirectly, not only death of AMs but also adverse alterations in their morphology and function, although some of the effects seemed to be reversible. Because AMs are crucial to the host against airborne pathogens, PRRSV infection may potentially predispose pigs to secondary pulmonary infections. PMID- 10665531 TI - Transmission of ovine herpesvirus 2 among adult sheep. AB - Previous studies from this laboratory have defined the pattern of acquisition of ovine herpesvirus 2 (OHV-2) in lambs under natural flock conditions. This study examined the question of whether OHV-2 could be transmitted between adult sheep. Two potential routes of transmission were examined: (1) direct inoculation of either viable leukocytes or whole blood from OHV-2 positive sheep, and (2) horizontal transmission through natural contact with OHV-2 positive sheep. Two groups of OHV-2 negative adult sheep were inoculated with material from infected sheep, one with 5x10(8) viable peripheral blood leukocytes (PBL), and the other with 100 ml of whole peripheral blood. No PCR signals were detected in any of the three sheep inoculated with the PBL during the 20 weeks following inoculation. In the group of five sheep inoculated with whole blood, two became PCR-positive at 7 and 8 weeks post-inoculation, respectively, and the remaining three sheep maintained their negative status until termination of the experiment at 20 weeks post-inoculation. In two experiments conducted in different flocks, a total of 20 adult sheep were used to examine horizontal transmission by contact; all animals became PCR-positive within 12 months of mixing the uninfected and infected animals. The results of these experiments support two conclusions. First, the susceptibility to OHV-2 is not limited to young lambs; adult sheep remain fully susceptible. Second, the fact that whole blood, but not PBL, from infected sheep was able to transmit the infection to only two of five inoculated sheep suggests that the infection in peripheral blood cells may be largely non-productive. PMID- 10665532 TI - Development and validation of a monoclonal antibody blocking ELISA for the detection of antibodies against both equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4). AB - A monoclonal antibody blocking ELISA was developed for the detection of antibodies directed against either EHV1 or EHV4. For this purpose, we selected a monoclonal antibody directed against a cross-reactive, conservative and immunodominant epitope of both EHV1 and EHV4. High antibody titres were found in rabbit antisera and SPF-foal antisera infected with either EHV1 or EHV4. After experimental challenge of conventional horses with EHV1 or EHV4 significant increases in CF and ELISA titres were found, whereas VN antibodies did not always increase significantly. In 344 paired serum samples submitted for diagnostic purposes a good agreement (kappa = 0.75, confidence limits = 0.63-0.88) was found between VN test and ELISA regarding a significant increase in titres. Also, a good correlation was found between VN and ELISA titres (r = 0.76, p<<0.0005). The relative sensitivity and specificity of the Mab blocking ELISA as compared with the VN test were 99.9 and 71%, respectively. The rather low relative specificity of the ELISA may be explained by a relatively low sensitivity of the VN test. The ELISA also detected increases in titre after vaccination with an EHV1 subunit vaccine, and after primary field infections in weaned foals. We concluded that the Mab blocking ELISA is more sensitive, easier to perform, more rapid and more reproducible than the VN test. We consider this test as a valuable tool for serological diagnosis of both EHV1 and EHV4 infections. PMID- 10665533 TI - Phylogenetic relationships of Staphylococcus aureus from bovine mastitis based on coagulase gene polymorphism. AB - Coagulase gene restriction fragment length polymorphism (RFLP) patterns were analyzed to determine the phylogenetic relationship among isolates of Staphylococcus aureus from the Czech Republic (n = 27), France (n = 48), Korea (n = 115) and the United States (n = 278). A total of 468 isolates of S. aureus were subtyped into 41 coagulase genotypes. Cluster analysis placed the 41 types into nine clusters. Eighteen API Staph profiles were determined for 102 S. aureus isolates representing 1 to 4 isolates of each coagulase type. The results of the study suggest that based on coagulase gene RFLP analysis, several genetic variants of S. aureus are prevalent. Comparison of coagulase and API Staph profiles indicated that the two identification system were independent of each other. PMID- 10665534 TI - Isolation of Campylobacter species from zoo animals and polymerase chain reaction based random amplified polymorphism DNA analysis. AB - A total of 104 fecal specimens from 30 mammals, 12 birds, and 3 reptiles at the Phoenix Zoological Gardens, Miyazaki City, Japan, were examined for the presence of Campylobacter species. All the animals examined were healthy with no fecal abnormality. Twenty-three (22.1%) thermophilic campylobacters, (9 C. jejuni, 11 C. hyointestinalis, 2 C. coli, and 1 C. lari), were isolated from 11 animals (7 mammals and 4 birds). C. jejuni and C. hyointestinalis were the predominant species isolated from these zoo animals and C. hyointestinalis was isolated frequently from simians. As selective media influence the numbers and species of campylobacters isolated, the agar medium was not supplemented with cephalothin. Campylobacters were isolated most frequently when a combination of enrichment culture and selective agar plating was performed at 42 degrees C. For the epidemiological study, a polymerase chain reaction (PCR)-based randomly amplified polymorphic DNA (RAPD) method was used as a tool to detect the heterogeneity of amplified DNAs of Campylobacter spp. isolated from zoo animals. The two arbitrary primers used in this study enabled even closely related strains of the same Campylobacter spp. to be differentiated. RAPD analysis revealed considerable diversity among the strains, suggesting that the transmission of Campylobacter spp. among animals in a defined area occurred through different mechanisms. Examination of the genotypic diversity among the multiple clones from the same host also revealed differences between clones. These results demonstrate that campylobacter populations in zoo animals are highly divergent. PMID- 10665535 TI - The association of two recombinant proteinases of a feline strain of Porphyromonas gingivalis with periodontal disease in cats. AB - Serum from 40 domestic cats with various grades of periodontal disease was used to probe two recombinant functional proteinases from feline strain VPB 3457 of Porphyromonas gingivalis expressed in E. coli. One recombinant proteinase (VPB 2856) was constructed using polymerase chain reaction and had 91% DNA identity with the prtC collagenase gene of the human type strain of P. gingivalis, while the other proteinase (VPB 2814) was isolated from a size selected genomic library and had an amino-terminal sequence with no significant identity with deposited sequences. Thirteen of 40 cats showed a serum antibody response to VPB 2856 using Western immunoblot detection. All the 13 cats had an overall periodontal grade of 3 or greater and greater than 1.68x10(5) cfu P. gingivalis at the canine and premolar periodontium sample sites. Fourteen of 40 cats showed a serum antibody response to VPB 2814. Thirteen of these 14 cats had an overall periodontal grade of 3 or greater. Regression analysis of overall periodontal grade against the serum antibody response showed significant positive relationships for both VPB 2856 (r2 = 0.351; p<0.001) and VPB 2814 (r2 = 0.247; p<0.001). Regression analysis of the total colony forming units of feline strain P. gingivalis against the grade of serum antibody response showed a positive relationship for both VPB 2856 (r2 = 0.662; p<0.001) and VPB 2814 (r2 = 0.531; p<0.001). These data provide strong evidence that the recombinant proteinases of feline P. gingivalis expressed in E. coli clones VPB 2856 and VPB 2814 are associated with periodontal disease in cats. PMID- 10665536 TI - Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae. AB - The reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae express one or two of three different RTX exotoxins designated Apx I, Apx II and Apx III. The toxins are important virulence factors. In the present study, ELISAs with purified Apx I, Apx II and Apx III, respectively, as antigen were evaluated as candidates for serological diagnosis of Actinobacillus pleuropneumoniae infection in pigs. The pigs were inoculated with biotype 1, serotypes 1-12, and biotype 2, serotype 14, respectively. A strong humoral antibody response was seen to all the three antigens in most pigs irrespective of the serotype used for inoculation. However, titers to the exotoxins secreted by the serotype used for inoculation were generally highest. The results show that toxin proteins of Actinobacillus pleuropneumoniae are antigenically related and that a correlation between serotype and secretion of exotoxin is not revealed serologically in the ELISA test. PMID- 10665537 TI - The dynamics of Staphylococcus aureus intramammary infection in nine Danish dairy herds. AB - The aim of the present study was to examine the diversity of Staphylococcus aureus isolates from bovine intramammary infections (IMI) in nine dairy herds, and compare these with isolates from other sites on the cows by phage- and ribotyping. Whether colonisation of milkers with S. aureus could be a source of infection for bovine IMI was investigated. In addition, 100 epidemiologically unrelated S. aureus isolates from asymptomatic human carriers were also phage- and ribotyped to compare the human and bovine reservoir of S. aureus in Denmark. A total of 625 S. aureus isolates from bovine IMI, bovine skin lesions, milking personnel, and non-farm-related human carriers were included in the study. Certain types predominated in one or several herds during the study period of one and-a-half to two years, whereas the presence of other types was of a more sporadic nature. Within the individual herds, there was a close correspondence between ribo- and phage types of S. aureus isolated from bovine intramammary infections and skin lesions. Isolates from milking personnel, however, were not identical to any of the predominant intramammary strains. Furthermore, several of the isolates from milking personnel showed ribo- and phage patterns identical to S. aureus isolates from human carriers. The findings of the present study underline the importance of strict milking hygiene and improvement of current mastitis therapy. The results support the hypothesis that some S. aureus mastitis strains are more contagious, virulent or persistent than others. The human reservoir of S. aureus does not play a major role as a source of bovine intramammary infections. PMID- 10665538 TI - Genomic fingerprinting of pigeon Streptococcus gallolyticus strains of different virulence by randomly amplified polymorphic DNA (RAPD) analysis. AB - Randomly amplified polymorphic DNA (RAPD) analysis was performed on 95 pigeon S. gallolyticus strains of different virulence and belonging to different biotypes and different culture supernatant phenotypes as determined by SDS-PAGE. Four distinct RAPD patterns, designated A, B, C and D, were distinguished using primer OPM6 (5'CTGGGCAACT). All 76 strains generating RAPD pattern A or B were designated highly virulent on the basis of their SDS-PAGE pattern. Five of seven strains generating RAPD pattern C and 11 of 12 strains generating RAPD pattern D belonged to the moderately virulent and low virulent culture supernatant phenotype groups, respectively. Only one RAPD group C strain belonged to a highly virulent culture supernatant phenotype group. There was a correlation between biotype and RAPD patterns. These findings indicate that there is a high correlation between RAPD pattern and virulence for pigeons. Therefore, RAPD typing seems a rapid, reliable method to distinguish pigeon S. gallolyticus strains of high, moderate and low virulence. PMID- 10665539 TI - Extensive diversity in New Zealand Dichelobacter nodosus strains from infected sheep and goats. AB - Footrot is a contagious bacterial disease of ruminants spread by the Gram negative, anaerobic organism, Dichelobacter nodosus. It is endemic in New Zealand and throughout sheep and goat farming regions of the world. Using the polymerase chain reaction (PCR) to amplify fragments of the fimbrial gene (fimA), D. nodosus was detected in 14 hoof scrapings, sampled from six farming regions within New Zealand. DNA sequencing revealed 15 strains covering eight serogroups on the New Zealand farms. The predominant serogroup was B which contained six strains, followed by serogroups F, H and G. No strains from serogroups D and I were detected in this investigation. Eleven out of the 15 D. nodosus strains had fimbriae sequences different to those previously reported and the presence of multiple strains on a single hoof was common (86% samples). Individual sheep from the same farm, or the same paddock, were often infected by a different range of strains, which suggests a host role in mediating footrot infection. PMID- 10665540 TI - A longitudinal study of Shiga-toxigenic Escherichia coli (STEC) prevalence in three Australian diary herds. AB - Over a 12 month period, 588 cattle faecal samples and 147 farm environmental samples from three dairy farms in southeast Queensland were examined for the presence of Shiga-toxigenic Escherichia coli (STEC). Samples were screened for Shiga toxin gene (stx) using PCR. Samples positive for stx were filtered onto hydrophobic grid membrane filters and STEC identified and isolated using colony hybridisation with a stx-specific DNA probe. Serotyping was performed to identify serogroups commonly associated with human infection or enterohaemorrhagic Escherichia coli (EHEC). Shiga-toxigenic Escherichia coli were isolated from 16.7% of cattle faecal samples and 4.1% of environmental samples. Of cattle STEC isolates, 10.2% serotyped as E. coli O26:H11 and 11.2% serotyped as E. coli O157:H7, and the E. coli O26:H11 and E. coli O157:H7 prevalences in the cattle samples were 1.7 and 1.9%, respectively. Prevalences for STEC and EHEC in dairy cattle faeces were similar to those derived in surveys within the northern and southern hemispheres. Calves at weaning were identified as the cattle group most likely to be shedding STEC, E. coli O26 or E. coli O157. In concurrence with previous studies, it appears that cattle, and in particular 1-14-week-old weanling calves, are the primary reservoir for STEC and EHEC on the dairy farm. PMID- 10665541 TI - Evaluation of a 23S rDNA polymerase chain reaction assay for identification of Serpulina intermedia, and strain typing using pulsed-field gel electrophoresis. AB - A polymerase chain reaction assay, amplifying a 1027 base pair portion of the 23S rDNA gene, was evaluated for identification of the intestinal spirochaete Serpulina intermedia. A total of 34 strains of S. intermedia isolated from pigs and chickens and 195 strains of other related species were tested. The optimised assay correctly identified all the S. intermedia strains, but generated 11 false positive reactions, giving a test sensitivity of 100% and a test specificity of 94.3%. The false positive reactions were generated from strains of four different species of intestinal spirochaetes, and the product was of the original predicted size. This suggests that the primer sites selected on the 23S rRNA gene were not completely specific for S. intermedia. Pulsed-field gel electrophoresis was then developed to investigate diversity amongst the S. intermedia strains. All strains tested had distinct DNA banding patterns using Mlu1, although three isolates from chickens on the same farm appeared closely related. The collection exhibited considerable genetic diversity, and strains from pigs and chickens were distributed in clusters throughout the dendrogram produced. The most closely related porcine and avian strains shared only 62% similarity. PMID- 10665542 TI - Monoclonal antibodies suitable for incorporation into a competitive enzyme-linked immunosorbent assay (ELISA) for detection of specific antibodies to Leptospira interrogans serovar pomona. AB - Monoclonal antibodies (mAb) were produced by fusing Sp2/0-Ag14 myeloma cells with spleen cells from BALB/c and ND4 mice that were immunized with killed Leptospira interrogans serovar pomona whole cells. Thirty hybridomas which produced antibodies (of the IgG1, IgG2a, IgG2b, or IgG3 isotype) that bound to epitopes on the serovar pomona whole cell antigen were identified by an indirect enzyme linked immunosorbent assay (ELISA). Twenty-eight of these 30 mAbs cross-reacted in the indirect ELISA with at least one whole cell antigen prepared from 12 other pathogenic Leptospira serovars, and/or with whole cell antigen from the non pathogenic Leptospira biflexa serovar patoc. The two serovar pomona-specific mAbs, which were designated M897 and M898, were obtained from the ND4 mouse and were both of the IgG1 isotype. In competitive ELISAs, M897 and M898 were inhibited from binding to the pomona antigen by bovine sera with anti-serovar pomona microscopic agglutination test (MAT) titres ranging from 100 to 6400. No significant inhibition was observed with pomona MAT-negative sera or with sera from animals experimentally infected with serovars canicola, copenhageni, grippotyphosa, hardjo type hardjobovis or sejroe. The epitopes recognized by M897 and M898 were both highly susceptible to sodium meta-periodate oxidation, indicating a carbohydrate composition. Neither of these mAbs reacted in immunoblots with the separated components of the serovar pomona whole cell antigen. PMID- 10665543 TI - Evaluation of three Brucella soluble antigens used in an indirect Elisa to discriminate S19 vaccinated from naturally infected cattle. AB - An O-polysaccharide (O-chain) and a hot-water extracted polysaccharide (PS), both obtained from Brucella abortus 1119-3, and a B. melitensis 16M native hapten (NH) were evaluated by indirect enzyme linked immunosorbent assay (ELISA) on three groups of cattle sera. The sera tested were: (a) 75 sera from cows naturally infected with B. abortus; (b) 130 sera from non-infected and non-vaccinated cattle; and (c) 61 sera from non-infected heifers recently vaccinated with B. abortus Strain 19 (S19). Sensitivity (Se), specificity (Sp) and the capability to discriminate vaccinated cattle (ADV) were determined. Using PS antigen, Se was 100% and the Sp was 97.7%, while the highest Sp was obtained by using the O-chain (99.2% ). For the NH antigen, Se was 94.7% and the Sp was 90.0%. The ADV of the three antigens was approximately 85%. Statistical analysis showed significant differences between O-chain/PS and O-chain/NH antigens. The agreement among antigens determined by kappa coefficient was 0.899 for O-chain/PS, 0.845 for O chain/NH and 0.795 for PS/NH. PMID- 10665544 TI - A metalloprotease is common to swine, avian and bovine isolates of Staphylococcus hyicus. AB - Staphylococcus hyicus is considered to be an etiological agent of exudative epidermitis in young pigs, but is frequently isolated from chickens and cows. In the present study, the proteases of 58 S. hyicus isolates from pigs, chickens and cows were examined by skim milk agar plate culture, gelatinolytic zymogram and polymerase chain reaction (PCR). These isolates showed proteolytic activity on skim milk agar plate, but activity differed amongst the isolates. In the gelatinolytic zymogram, one main band was observed in all the porcine, avian and bovine isolates, while one to two other bands were recognized in some isolates. The formation of the main band was inhibited by EDTA, suggesting that this protease is a metalloprotease. When the Shp1 gene, which codes for one the metalloproteases of S. hyicus as reported previously, was examined by PCR, one band arising from an open reading frame (ORF) was detected in all of 58 isolates tested. In addition, upstream nucleotides containing the promoter region of Shp1 gene were amplified and sequenced. From these results, it seems likely that the metalloprotease is common to porcine, avian and bovine isolates of S. hyicus. PMID- 10665545 TI - Gene therapy under cloud. PMID- 10665546 TI - Difficulties with oral platelet glycoprotein IIb/IIIa receptor antagonists. PMID- 10665547 TI - Prevention of transfusion-transmitted hepatitis. PMID- 10665548 TI - Transmission of Helicobacter pylori: is it all child's play? PMID- 10665549 TI - Antithrombotic drug therapy after infrainguinal vascular surgery. PMID- 10665550 TI - Seizures and epilepsy after traumatic brain injury. PMID- 10665551 TI - UK and US health-care systems: divided by more than a common language. PMID- 10665552 TI - Comparison of sibrafiban with aspirin for prevention of cardiovascular events after acute coronary syndromes: a randomised trial. The SYMPHONY Investigators. Sibrafiban versus Aspirin to Yield Maximum Protection from Ischemic Heart Events Post-acute Coronary Syndromes. AB - BACKGROUND: Aspirin lowers risks of death and myocardial infarction in patients with acute coronary syndromes. Intravenous glycoprotein IIb/IIIa receptor antagonists further reduce the rates of ischaemic events in these patients, but the efficacy of long-term oral glycoprotein IIb/IIIa receptor blockade has not been established. We tested whether the oral glycoprotein IIb/IIIa receptor antagonist sibrafiban would prevent more cardiovascular events than aspirin, when given within 7 days of, and sustained for 90 days after, an acute coronary syndrome event. METHODS: 9233 patients who had stabilised after an acute coronary syndrome event were randomly assigned aspirin (80 mg orally twice daily) or low dose or high-dose sibrafiban. Sibrafiban doses (3.0 mg, 4.5 mg, or 6.0 mg) were based on a model accounting for weight and serum creatinine and designed to achieve at least 25% steady-state inhibition of platelet aggregation (low dose) or at least 50% inhibition (high dose). The primary endpoint was the composite of death, non-fatal infarction or reinfarction, or severe recurrent ischaemia at 90 days. Analysis was by intention to treat. FINDINGS: The 90-day rate of the primary endpoint did not differ significantly between the groups assigned aspirin (302 [9.8%]), low-dose sibrafiban (310 [10.1%]; odds ratio 1.03 [95% CI 0.87 1.21]), and high-dose sibrafiban (303 [10.1%]; 1.03 [0.87-1.21]). The groups did not differ significantly in the rates of the component events or secondary efficacy endpoints. Major bleeding was more common with high-dose sibrafiban (171 [5.7%]) than with aspirin (120 [3.9%]) or low-dose sibrafiban (159 [5.2%]). INTERPRETATION: Sibrafiban showed no additional benefit over aspirin for secondary prevention of major ischaemic events after an acute coronary syndrome, and was associated with more dose-related bleeding. PMID- 10665553 TI - Efficacy of oral anticoagulants compared with aspirin after infrainguinal bypass surgery (The Dutch Bypass Oral Anticoagulants or Aspirin Study): a randomised trial. AB - BACKGROUND: Oral anticoagulants and aspirin are antithrombotic drugs that are commonly used in patients with vascular disease. We investigated whether either of these treatments prevented more effectively than the other bypass complications after infrainguinal bypass surgery. METHODS: We did a multicentre, randomised, open trial. 2690 patients who had undergone infrainguinal grafting were randomly assigned oral anticoagulants (target international normalised ratio 3.0-4.5, n=1339) or aspirin (80 mg daily, n=1351). We followed up patients for a mean of 21 months. The primary outcome was graft occlusion. FINDINGS: 308 graft occlusions occurred in the oral-anticoagulants group compared with 322 in the aspirin group (hazard ratio 0.95 [95% CI 0.82-1.11]), which suggested no overall advantage for either treatment. Oral anticoagulants were beneficial in patients with vein grafts (0.69 [0.54-0.88]), whereas aspirin had better results for nonvenous grafts (1.26 [1.03-1.55]). The composite outcome of vascular death, myocardial infarction, stroke, or amputation occurred 248 times in the oral anticoagulants group and 275 times in the aspirin group (0.89 [0.75-1.06]). Patients treated with oral anticoagulants had more major bleeding episodes than those treated with aspirin (108 vs 56; 1.96 [1.42-2.71]). INTERPRETATION: Oral anticoagulation was better for the prevention of infrainguinal-vein-graft occlusion and for lowering the rate of ischaemic events. Aspirin was better for the prevention of non-venous graft occlusion, and was associated with fewer bleeding episodes. PMID- 10665554 TI - Efficacy of artesunate plus pyrimethamine-sulphadoxine for uncomplicated malaria in Gambian children: a double-blind, randomised, controlled trial. AB - BACKGROUND: Resistance to cheap effective antimalarial drugs, especially to pyrimethaminesulphadoxine (Fansidar), is likely to have a striking impact on childhood mortality in sub-Sharan Africa. The use of artesunate (artesunic acid) [corrected] in combination with pyrimethamine-sulphadoxine may delay or prevent resistance. We investigated the efficacy, safety, and tolerability of this combined treatment. METHODS: We did a double-blind, randomised, placebo controlled trial in The Gambia. 600 children with acute uncomplicated Plasmodium falciparum malaria, aged 6 months to 10 years, at five health centres were randomly assigned pyrimethaminesulphadoxine (25 mg/500 mg) with placebo; pyrimethamine-sulphadoxine plus one dose of artesunate (4mg/kg bodyweight); or pyrimethamine-sulphadoxine plus one dose 4 mg/kg bodyweight artesunate daily for 3 days. Children were visited at home each day after the start of treatment until parasitaemia had cleared. FINDINGS: The combined treatment was well tolerated. No adverse reactions attributable to treatment were recorded. By day 1, only 178 (47%) of 381 children treated with artesunate were still parasitaemic, compared with 157 (81%) of 195 children in the pyrimethamine-sulphadoxine alone group (relative risk 1.7 [95% CI 1.5-2.0], p<0.001). Treatment-failure rates at day 14 were 3.1% in the pyrimethamine sulphadoxine alone group, and 3.7% in the one-dose artesunate group (risk difference -0.6% [-4.2 to 3.0]) and 1.6% in the three-dose group (1.5 [1.5-4.5], p=0.048). Symptoms resolved faster in children who received artesunate, but there was no additional benefit for three doses of artesunate over one dose. Children given artesunate were less likely to be gametocytaemic after treatment. INTERPRETATION: The combined treatment was safe, well tolerated, and effective. The addition of artesunate to malaria treatment regimens in Africa results in lower gametocyte rates and may lower transmission rates. PMID- 10665555 TI - Transmission of Helicobacter pylori among siblings. AB - BACKGROUND: Helicobacter pylori infection causes chronic digestive diseases with a major public-health impact, yet the design of prevention measures is hampered by limited knowledge of transmission pathways. We studied the effect of family composition on H. pylori prevalence among rural Colombian children aged 2-9 years. METHODS: 684 children were screened for H. pylori with the 13C-urea breath test. For each child, birth order, birth spacing, number of 2-9-year-old siblings, and number of H-pylori-positive 2-9-year-old siblings was recorded. Odds ratios were estimated by logistic regression, controlling for hygiene related exposures, socioeconomic indicators, and the number of children in the household. FINDINGS: The odds of infection increased with the number of 2-9-year old siblings in the household (odds ratios 1.4, 2.3, 2.6, and 4.3 for one, two, three, and four to five siblings, respectively). Compared with first-born children, odds ratios for children born second and third to ninth were 1.8 (95% CI 1.0-3.3) and 2.2 (1.0-4.3), respectively. Compared with children born 10 or more years after the next older household member, those born within 4 years were 4.1 times (CI 2.0-8.6) more likely to be infected; the age gap to the next younger household member displayed a weaker effect. The number of H-pylori positive 2-9-year-old siblings had a particularly strong effect gradient (1.5, 3.2, 5.6, and 7.1, for one, two, three, and four positive siblings, respectively). INTERPRETATION: Among rural Andean children younger than 10 years, H. pylori infection seems to be transmitted most readily among siblings who are close in age, and most frequently from older siblings to younger ones. PMID- 10665556 TI - Results at 1 year of outpatient multidisciplinary pulmonary rehabilitation: a randomised controlled trial. AB - BACKGROUND: Pulmonary rehabilitation seems to be an effective intervention in patients with chronic obstructive pulmonary disease. We undertook a randomised controlled trial to assess the effect of outpatient pulmonary rehabilitation on use of health care and patients' wellbeing over 1 year. METHODS: 200 patients with disabling chronic lung disease (the majority with chronic obstructive pulmonary disease) were randomly assigned a 6-week multidisciplinary rehabilitation programme (18 visits) or standard medical management. Use of health services was assessed from hospital and general-practice records. Analysis was by intention to treat. FINDINGS: There was no difference between the rehabilitation (n=99) and control (n=101) groups in the number of patients admitted to hospital (40 vs 41) but the number of days these patients spent in hospital differed significantly (mean 10.4 [SD 9.7] vs 21.0 [20.7], p=0.022). The rehabilitation group had more primary-care consultations at the general practitioner's premises than did the control group (8.6 [6.8] vs 7.3 [8.3], p=0.033) but fewer primary-care home visits (1.5 [2.8] vs 2.8 [4.6], p=0.037). Compared with control, the rehabilitation group also showed greater improvements in walking ability and in general and disease-specific health status. INTERPRETATION: For patients chronically disabled by obstructive pulmonary disease, an intensive, multidisciplinary, outpatient programme of rehabilitation is an effective intervention, in the short term and the long term, that decreases use of health services. PMID- 10665557 TI - Clinical presentation and outcome of Pneumocystis carinii pneumonia in Malawian children. AB - BACKGROUND: Necropsy studies from Africa have shown that Pneumocystis carinii pneumonia (PCP) is common in infants with HIV infection. We aimed to describe the rate, clinical presentation, and outcome of PCP in young Malawian children with acute severe pneumonia. METHODS: Children aged between 2 months and 5 years who were in hospital with a diagnosis of severe pneumonia were admitted to a study ward for clinical monitoring. We carried out blood culture, immunofluorescence on nasopharyngeal aspirate samples to test for PCP, polymerase chain reaction to detect HIV, and chest radiography. FINDINGS: 16 cases of PCP were identified among 150 children with radiologically confirmed severe pneumonia. All were HIV positive and younger than 6 months. 21 children had bacterial pneumonia (including one who was also PCP positive) and 114 were not confirmed. The most common bacterial pathogens among children without PCP were Streptococcus pneumoniae (eight) and non-typhoidal salmonellae (seven). On admission, children with confirmed PCP had a lower mean age, body temperature, and oxygen saturation than children with bacterial pneumonia and were less likely to have a focal abnormality on auscultation. Oxygen requirements were much greater in children with PCP than those with bacterial pneumonias (96 of 105 hospital days vs 15 of 94, p<0.0001). Ten of 16 children with PCP and six of 21 with bacterial pneumonia died (relative risk 2.19 [95% CI 1.0-4.7]). The overall case-fatality rate of severe pneumonia was 22%. In addition to a strong association with PCP, a fatal outcome was significantly and independently associated with HIV infection (2.98 [1.1-7.9]) and with age under 6 months (2.76 [1.0-5.2]). INTERPRETATION: PCP is common and contributes to the high mortality from pneumonia in Malawian infants. Clinical features are helpful in diagnosis. The study highlights the impact of HIV infection and difficult issues of management in countries with few resources. PMID- 10665558 TI - Confused by arteritis. PMID- 10665559 TI - Resuscitation from accidental hypothermia of 13.7 degrees C with circulatory arrest. AB - In a victim of very deep accidental hypothermia, 9 h of resuscitation and stabilisation led to good physical and mental recovery. This potential outcome should be borne in mind for all such victims. PMID- 10665560 TI - Sirolimus-tacrolimus combination immunosuppression. AB - A series of 32 recipients of liver, kidney, or pancreas transplants who were treated with sirolimus and low-dose tacrolimus experienced a low rate of rejection and excellent graft function without drug-related toxic effects. PMID- 10665561 TI - Helicobacter pylori and epidemic Vibrio cholerae O1 infection in Peru. AB - In a cross-sectional study of the 1991 Peruvian cholera epidemic, Vibrio cholerae O1 infection was associated with Helicobacter pylori infection, particularly in young children. These data support the hypothesis that hypochlorhydria induced by H. pylori is important in the pathogenesis of diarrhoeal disease. PMID- 10665562 TI - Inflammatory bowel disease in non-Ashkenazi Jews with familial Mediterranean fever. AB - Familial Mediterranean fever and inflammatory bowel disease are two inflammatory conditions. We showed that inflammatory bowel disease was particularly frequent and severe in non-Ashkenazi Jewish patients with familial Mediterranean fever. PMID- 10665563 TI - Dopamine-transporter density in nucleus accumbens of type-1 alcoholics. AB - We have shown that alcoholic patients have a lower number of dopamine transporters in the nucleus accumbens, which mediates the rewarding effects of addictive drugs. Thus, certain dopaminergic agents may be beneficial in the treatment of alcohol withdrawal and in the long-term treatment of alcoholism with selective use. PMID- 10665564 TI - Earliest stages of tumour-induced angiogenesis dissected. PMID- 10665565 TI - The balance of risk and benefit in gene-therapy trials. PMID- 10665567 TI - WHO assembles leading economists to study poverty reduction and health. PMID- 10665566 TI - At last, the fight against lymphatic filiariasis begins. PMID- 10665568 TI - UK allows frozen eggs for fertility treatment. PMID- 10665569 TI - Mitochondrial respiratory chain disorders II: neurodegenerative disorders and nuclear gene defects. AB - The first part of this review (Lancet 2000; 355: 299) covered primary disorders of mitochondrial DNA (mtDNA). This section will cover nuclear-encoded defects of the oxidative phosphorylation (OXPHOS) system, including mtDNA mutations that are secondary to nuclear gene mutations and nuclear gene defects responsible for secondary OXPHOS deficiency (panel). The latter group of diseases are predominantly neurodegenerative. The mitochondrion's role in apoptosis and its contribution to the pathogenesis of neurodegenerative diseases are also covered. PMID- 10665570 TI - Surveillance after colorectal cancer resection. PMID- 10665571 TI - Condoms and seat belts: the parallels and the lessons. PMID- 10665572 TI - Psychological response and survival in breast cancer. PMID- 10665573 TI - Psychological response and survival in breast cancer. PMID- 10665574 TI - Psychological response and survival in breast cancer. PMID- 10665575 TI - Psychological response and survival in breast cancer. PMID- 10665576 TI - Psychological response and survival in breast cancer. PMID- 10665577 TI - Prophylactic antidepressant treatment before patients are admitted. PMID- 10665578 TI - Hyperemesis gravidarum and sex of child. PMID- 10665579 TI - Hyperemesis gravidarum and sex of child. PMID- 10665580 TI - Xenotransplantation debate. PMID- 10665581 TI - Xenotransplantation debate. PMID- 10665582 TI - Autism and measles, mumps, and rubella vaccine. PMID- 10665583 TI - Cervical screening. PMID- 10665584 TI - Cervical screening. PMID- 10665585 TI - Cervical screening. PMID- 10665586 TI - Cervical screening. PMID- 10665587 TI - Polycystic ovarian disease: a misleading label? PMID- 10665588 TI - Aortic pulse-wave velocity versus pulse pressure and pulse-wave analysis. PMID- 10665589 TI - General anaesthesia in dental treatment. PMID- 10665590 TI - Bias against publication of surgical papers. PMID- 10665591 TI - Probiotics strain for credibility. PMID- 10665592 TI - Do genetically modified foods affect human health? PMID- 10665593 TI - Trap-door medicine. PMID- 10665594 TI - Linus Pauling: an inspirational and humane genius. PMID- 10665595 TI - The Nobel chronicles. 1985: Joseph Leonard Goldstein (b 1940), Michael Stuart Brown (b 1941). PMID- 10665596 TI - Testing adolescents for a hereditary breast cancer gene (BRCA1): respecting their autonomy is in their best interest. AB - The testing of individuals before the age of 18 years for hereditary late-onset diseases has been judged ethically not acceptable in guidelines and directives published by medical professional organizations. However, there are not enough best interest arguments to deny genetic testing to an adolescent at risk of carrying a BRCA1 mutation, even if the competence of adolescents for medical decisions is considered to be lower than the competence of adults. The adolescent's decision is not irrational or of very high risk. Respecting adolescents' autonomous choices concerning genetic testing has positive consequences for their self-esteem and psychological health. Geneticists and other professionals should clearly differentiate between children and adolescents in regard to BRCA1 gene testing and recommend giving decision autonomy about the test to all psychologically "normal" adolescents. PMID- 10665597 TI - Subspecialty referrals for pauciarticular juvenile rheumatoid arthritis. AB - OBJECTIVES: To examine referral patterns from primary care physicians for children with pauciarticular juvenile rheumatoid arthritis (JRA) and to determine whether children with pauciarticular JRA referred to pediatric rheumatologists differ in clinical presentation from children referred to other specialists. DESIGN: A retrospective records review of 49 patients with pauciarticular jRA was performed. Records were reviewed to determine the specialty of the referring physician and whether the children referred had symptoms and signs compatible with a synovitis at the time primary care was sought. SETTING: Inner-city tertiary pediatric rheumatology referral center. PARTICIPANTS: Children with pauciarticular JRA. MAIN OUTCOME MEASURES: Identification of referral patterns of primary care physicians. Associated morbidity owing to JRA was ascertained at the time of referral. RESULTS: Most children with pauciarticular JRA (62%) were referred to orthopedic surgeons prior to referral for pediatric rheumatology care. No differences in clinical symptoms were seen between children referred to pediatric rheumatologists and those referred to orthopedic surgeons. Children referred initially to orthopedic surgeons were younger than those referred to pediatric rheumatologists. CONCLUSION: A notable number of children with pauciarticular JRA are referred to orthopedic surgeons prior to the establishment of that diagnosis, even when such children present with unequivocal signs of synovitis. This may be owing to the misconception that arthritis is rare in preschool-aged children or to the difficulty of ascertaining the presence of synovitis in younger children. PMID- 10665598 TI - Prevalence of and risks for cervical human papillomavirus infection and squamous intraepithelial lesions in adolescent girls: impact of infection with human immunodeficiency virus. AB - CONTEXT: Data suggest that in adults, human papillomavirus (HPV) infections and their sequalae, squamous intraepithelial lesions (SILs), occur more commonly among human immunodeficiency (HIV)-infected women because of the HIV-associated CD4+ T-cell immunosuppression. Since adolescents are more likely to be early in the course of HIV and HPV infections, the study of both infections in this age group may help elucidate their initial relationship. OBJECTIVE: To examine the prevalence of and risks for cervical HPV infection and SILs by HIV status in a population of adolescent girls. PARTICIPANTS: Subjects recruited at each of the 16 different US sites participating in a national study of HIV infection in adolescents. MAIN OUTCOME MEASURES: Cervical HPV DNA findings using polymerase chain reaction detection techniques and Papanicolaou smear from baseline visits. Infection with HPV was categorized into low- (rarely associated with cancer) and high- (commonly associated with cancers) risk types. RESULTS: Of 133 HIV-infected girls, 103 (77.4%) compared with 30 (54.5%) of 55 noninfected girls were positive for HPV (relative risk [RR], 1.4; 95% confidence interval [CI], 1.1-1.8). The risk was for high-risk (RR, 1.8; 95% CI, 1.2-2.7) but not low-risk (RR, 1.2; 95% Cl, 0.4-3.9) HPV types. Among the girls with HPV infection, 21 (70.0%) of the non HIV-infected girls had normal cytologic findings compared with only 29 (29.9%) of the HIV-infected girls (P<.001). Multivariate analysis showed that HIV status was a significant risk for HPV infection (odds ratio [OR], 3.3; 95% CI, 1.6-6.7) and SIL (OR, 4.7; 95% CI, 1.8-14.8), but CD4 cell count and viral load were not associated with infection or squamous intraepithelial lesions. Only 9 girls had a CD4+ T-cell count of less than 0.2 cell X 10(9)/L. CONCLUSIONS: High prevalence of HPV infection in both groups underscores the risky sexual behavior in this adolescent cohort. Rates of HPV infection and SILs were higher among HIV-infected girls, despite similar sexual risk behaviors and the relatively healthy state of our HIV-infected group. Infection with HIV may enhance HPV proliferation through mechanisms other than CD4 immunosuppression, particularly early in the course of HIV infection. PMID- 10665599 TI - Fathers and child neglect. AB - OBJECTIVE: To examine the association between father involvement and child neglect. DESIGN: Cohort study. SETTING: Participants were recruited from an inner city pediatric primary care clinic and a clinic for children at risk for human immunodeficiency virus infection in a teaching hospital. PARTICIPANTS: Mothers and fathers or father figures, and 244 five-year olds participating in a longitudinal study. MAIN OUTCOME MEASURES: Child neglect measured via home observation, a videotaped mother-child interaction, and child protective services reports. RESULTS: A father or father figure was identified for 72% of the children. Rates of neglect ranged between 11% and 30%. Father absence alone was not associated with neglect. However, in families with an identified and interviewed father, a longer duration of involvement (P<.01), a greater sense of parenting efficacy (P<.01), more involvement with household tasks (P<.05), and less involvement with child care (P<.05) were associated with less neglect. The overall model explained 26.5% of the variance in neglect. CONCLUSIONS: There is substantial involvement of fathers in a subset of this high-risk sample, although more than a quarter of the children lacked a father or father figure. The mere presence of a father did not significantly influence the neglect of the children; rather, the nature of his involvement did. Fathers who felt more effective as parents were less likely to have neglected their children. A greater sense of efficacy may reflect parenting skills and be important in enhancing the contribution of fathers to their children's well-being. Pediatric health care providers can play a valuable role in enhancing the involvement and skills of fathers. PMID- 10665600 TI - Asthma severity among children hospitalized in 1990 and 1995. AB - BACKGROUND: During the past decade, the number of children with asthma increased; however, the number of asthma hospitalizations for children decreased. OBJECTIVE: To assess the proportion of high-severity cases among children hospitalized with asthma and the association of high-severity asthma with patient and hospital characteristics. DESIGN: The data set used was the Healthcare Cost and Utilization Project Nationwide Inpatient Sample. Records were selected of patients 18 years and younger who had the principal diagnosis of asthma. Records were analyzed of 29077 patients at 746 hospitals in 1990 and 33 443 patients at 811 hospitals in 1995. Asthma severity was classified by All Patient Refined Diagnosis-Related Groups. Cross-sectional logistic regression analysis was performed using survey data analysis software. RESULTS: The most common diagnoses associated with high-severity asthma were respiratory distress and respiratory failure. The proportion of high-severity asthma cases did not change significantly between 1990 (4.2%) and 1995 (4.6%) (P = .08). Adolescents and boys were more likely to have high-severity asthma than children aged 5 to 12 years and girls. Western, southern, and north-central hospitals and urban teaching hospitals had a higher proportion of high-severity asthma cases than northeastern hospitals and nonteaching hospitals. CONCLUSIONS: Between 1990 and 1995, the proportion of high-severity cases among children hospitalized with asthma did not change significantly. However, patient age, sex, region of the country, and hospital teaching status were associated with variations in the proportion of high-severity asthma cases. PMID- 10665601 TI - Prevalence of symptoms of gastroesophageal reflux during childhood: a pediatric practice-based survey. Pediatric Practice Research Group. AB - OBJECTIVES: To determine the prevalence of symptoms associated with gastroesophageal reflux (GER) in 3- to 17-year-old children, to describe the prevalence of factors associated with GER in these children, and to determine the percentage of symptomatic children who have been treated. DESIGN: A cross sectional survey. SETTING: Sixteen pediatric practice research group practices in the Chicago, Ill, area (urban, suburban, and semirural). PARTICIPANTS: A total of 566 parents of 3- to 9-year-old children, 584 parents of 10- to 17-year-old children, and 615 children aged 10 to 17 years. INTERVENTION: None. MAIN OUTCOME MEASURE: Reported frequency of symptoms associated with GER. RESULTS: Parents of 3- to 9-year-old children reported that their children experienced a sensation of heartburn ("burning/painful feeling in middle of chest"), epigastric pain ("stomachache above belly button"), and regurgitation ("sour taste or taste of throw up") 1.8%, 7.2%, and 2.3% of the time, respectively. Parents of 10- to 17 year-old children reported that their children experienced the same symptoms 3.5%, 3.0%, and 1.4% of the time, while children aged 10 to 17 years reported the symptoms 5.2%, 5.0%, and 8.2% of the time, respectively. Complaints of abdominal pain ("stomachache") were most common, reported by 23.9% and 14.7% of parents of 3- to 9-year-old and 10- to 17-year-old children and by 27.9% of children aged 10 to 17 years. In those aged 10 to 17 years, heartburn reported by the children was associated with reported cigarette use (odds ratio, 6.5; 95% confidence interval, 2-21); no other complaint was associated with cigarette, alcohol, or caffeine consumption or passive smoking exposure. In 3- to 9-year-old children, no complaint was associated with caffeine consumption or passive smoking exposure. Reported treatment in the past week with antacids was 0.5% according to parents of children aged 3 to 9 years and 1.9% and 2.3% according to parents of children aged 10 to 17 years and children aged 10 to 17 years, respectively. Treatment with over-the-counter histamine receptor blockers was 0% for children aged 3 to 9 years and 10 to 17 years, as reported by their parents, and 1.3% for those aged 10 to 17 years, as reported by themselves. CONCLUSIONS: Symptoms suggestive of GER are not rare in childhood, yet only a fraction of children with symptoms are treated with over-the-counter antacids or histamine2 antagonists. Prospective longitudinal data are needed to determine which children with symptoms of GER actually have GER disease and are at risk of developing complications. PMID- 10665602 TI - Secular trends in height among children during 2 decades: The Bogalusa Heart Study. AB - OBJECTIVE: To examine trends in height among 5- to 17-year-old children between 1973 and 1992. DESIGN: A panel design consisting of 7 cross-sectional surveys. PARTICIPANTS: All schoolchildren residing in Bogalusa, La, were eligible. A total of 24 070 examinations were performed. RESULTS: During the study period, the mean height of schoolchildren increased by 0.70 cm per decade independently of race, sex, and age. Trends were most pronounced among preadolescents, blacks, and boys, with 9- to 12-year-old black boys showing a height increase of 1.8 cm per decade. We observed a decrease in the number of relatively short children (<10th percentile of height) and an increase in the number of tall children (>90th percentile of height). Because a secular trend was not seen among the 15- to 17 year-old children, our findings likely reflect an acceleration of maturation. CONCLUSIONS: It has generally been assumed that secular increases in height among schoolchildren in the United States ceased by the mid-1900s. Our findings, which may be due to various environmental factors, demonstrate that care must be taken when using nonconcurrent reference data to assess the growth of children. Additional study is needed to determine if these secular trends are continuing and to examine possible explanations and consequences of these trends. PMID- 10665603 TI - Adolescent patients--healthy or hurting? Missed opportunities to screen for suicide risk in the primary care setting. AB - CONTEXT: Adolescent suicide rates have increased dramatically in recent decades. Suicide is the third leading cause of mortality among persons aged 10 to 19 years. Several official guidelines recommend screening for suicidal behavior in the primary care setting. OBJECTIVES: To determine the prevalence of adolescent suicidal behavior known to primary care providers and to determine the knowledge, attitudes, and practice of primary care physicians in Maryland regarding screening for risk factors for adolescent suicide. DESIGN: Cross-sectional study using mailed survey. SETTING: Maryland from May to July 1995. PARTICIPANTS: All pediatrician (n = 816) and family physician (n = 592) members of the state chapter of the American Academy of Pediatrics and the American Academy of Family Physicians, respectively, who were actively providing ambulatory care. MAIN OUTCOME MEASURES: Adolescent suicidal behavior known to primary care providers and predictors of routine screening for risk factors for adolescent suicide. RESULTS: The response rate was 66%. Three hundred twenty-eight physicians (47%) reported that 1 or more adolescent patients attempted suicide in the previous year, but only 158 (23%) either frequently or always screened adolescent patients for suicide risk factors. Significant factors correlating with routine screening for suicide risk factors included frequently or always counseling about the safer storage of firearms in the home (odds ratio [OR], 5.3; 95% confidence interval [CI], 2.8-10.2); agreeing or strongly agreeing that they were sufficiently trained and knew how to screen for risk factors (OR, 3.2; 95%/CI, 1.7-6.3); agreeing or strongly agreeing that they had enough time during the well visit to screen for mental health problems (OR, 2.9: 95% CI, 1.6-5.3); frequently or always counseling about child passenger safety (OR, 2.7; 95% CI, 1.6-4.7); spending more than 5 minutes in anticipatory guidance during the well visit (OR, 2.7: 95% CI, 1.5-4.6); practicing in an urban setting (OR, 2.3; 95)% CI, 1.2 4.7); agreeing or strongly agreeing that physicians can be effective in preventing adolescent suicide and that what they do during an office visit may help prevent adolescent suicide (OR, 2.0; 95% CI, 1.2-3.4); and female sex (OR. 1.9; 95% CI, 1.1-3.2). CONCLUSION: Despite the substantial proportion of primary care providers who encountered suicidal adolescent patients, most providers still do not routinely screen their patients for suicidality or associated risk factors. More training is needed and desired by the survey respondents. Patient confidentiality issues must be addressed. Development and widespread use of a short, easily administered, reliable, and valid screening tool are recommended to help busy clinicians obtain more complete information during all visits. PMID- 10665604 TI - Acute primary Chlamydia trachomatis infection in male adolescents after their first sexual contact. AB - BACKGROUND: Chlamydia trachomatis infection occurs primarily among youth sexually active persons. Few studies have evaluated the kinetics of markers of infection in male adolescents after their first sexual contact. DESIGN: Primary C trachomitis infection in 4 young male adolescents after their first sexual contact was diagnosed by polymerase chain reaction and antigen detection in sequential first voiding urine and urethral specimens, respectively. Serial serum samples were assessed for the presence of specific IgA and IgG antibodies. RESULTS: Both polymerase chain reaction and antigen detection correctly identified all cases of primary C trachomatis infection. The polymerase chain reaction method was, however, an earlier marker of infection. Three patients were seronegative at presentation. Two of these subsequently seroconverted to either IgA or IgG, while the third remains seronegative. The time interval from onset of symptoms to seroconversion ranged from 10 to 25 days. CONCLUSIONS: Although polymerase chain reaction and antigen and serologic detection have previously been described in primary C trachomatis infection, this report documents the variability of these markers during the first phase of infection in non-sexually active young male adolescents. C trachomatis can be acquired by male adolescents after their first sexual contact; however, there is a prolonged period when the patient is seronegative, yet infections can occur. PMID- 10665605 TI - Preventive services in a health maintenance organization: how well do pediatricians screen and educate adolescent patients? AB - OBJECTIVE: To determine whether pediatricians in managed care settings adhere to national guidelines concerning the provision of clinical preventive services. DESIGN: Surveys were mailed between September 1996 and April 1997 to all pediatricians practicing in a California group-model health maintenance organization. The survey asked pediatricians about their screening and education practices on 34 recommended services and the actions taken with adolescent patients who have engaged in risk behavior. RESULTS: The response rate was 66.2% (N = 366). Pediatricians, on average, screened 92% of their adolescent patients for immunization status and blood pressure; 85% for school performance; 60% to 80% for obesity, sexual intercourse, cigarette use, alcohol use, drug use, and seat belt and helmet use; 30% to 47% for access to handguns, suicide, eating disorders, depression, and driving after drinking alcohol; fewer than 20% for use of smokeless tobacco, sexual orientation, sexual and physical abuse, and riding a bike or swimming after drinking alcohol; and 26% to 41% for close friends' engagement in risk behavior. Pediatricians' assessment and education with adolescent patients who screened positive for risk behavior was particularly low. Female physicians, physicians who saw a greater proportion of older adolescents, and recent medical school graduates were more likely to provide preventive services. CONCLUSIONS: Pediatricians in this health maintenance organization provide preventive services to adolescent patients at rates below recommendations but at rates greater than physicians in other practice settings. Improvement is especially needed in the areas that contribute most to adolescent mortality and for patients who screen positive for a risk behavior. PMID- 10665606 TI - Child care center staff contribute to physician visits and pressure for antibiotic prescription. AB - OBJECTIVE: To determine whether child care center (CCC) providers contribute to unnecessary physician referrals and antibiotic prescriptions in young children with upper respiratory tract infections. DESIGN: A survey using a structured telephone questionnaire between May 3, 1998, and July 27, 1998. PARTICIPANTS: Child care center providers from randomly selected licensed Ontario CCCs accepting diapered children. MAIN OUTCOME MEASURES: Knowledge, attitudes, and practices concerning physician referral; exclusion; and antibiotic use for children with upper respiratory tract infections. Indications for exclusion were compared with published Canadian guidelines. RESULTS: Contact was made with 42 eligible CCCs to obtain the requisite number of 36 participants (participation rate, 86%). Of the 36 centers, staff reported advising that children visit a physician for colored nasal discharge in 28 (78%), for productive cough in 23 (64%), and for unusual behavior in 9 (25%). Also of the 36 centers, staff reported excluding children for colored nasal discharge in 20 (56%), for productive cough in 16 (44%), and for unusual behavior in 15 (42%). Antibiotics were thought useful for nonspecific upper respiratory tract infections to prevent the spread of infection in 9 (26%), to speed up recovery in 7 (21%), and to prevent bacterial infection in 13 (38%) of 34 centers. In the previous 6 months, 25 (69%) of 36 staff members reported making an exception to exclusion because a child had an antibiotic prescription. CONCLUSIONS: Many children are referred by CCC staff to physicians contrary to established guidelines. As staff must act on behalf of parents, a low threshold for referral is not unreasonable. However, this survey confirms that CCC staff recommend children to receive antibiotics and exclude children inappropriately. These practices are based on incomplete knowledge. Research on appropriate management of upper respiratory tract infections by CCC staff is needed. Education to correct specific knowledge deficits should be initiated. PMID- 10665607 TI - Vaccination practices, policies, and management factors associated with high vaccination coverage levels in Georgia public clinics. Georgia Immunization Program Evaluation Team. AB - BACKGROUND: Controlling vaccine-preventable diseases by achieving high childhood vaccination coverage levels is a national priority. However, there are few, if any, comprehensive evaluations of state immunization programs in the United States, and little attention has been given to the importance of vaccination clinic management style and staff motivation. OBJECTIVE: To evaluate the factors associated with the increase in childhood vaccination coverage levels from 53% in 1988 to 89% in 1994 in Georgia's public health clinics. DESIGN: A 1994 mail survey obtaining information on clinic vaccination policies and practices and management practices. SETTING: All 227 public health clinics in Georgia. PARTICIPANTS: Clinic nurses responsible for vaccination services. OUTCOME MEASURE: The 1994 clinic-specific coverage level for 21- to 23-month-old children for 4 doses of diphtheria and tetanus toxoids and pertussis vaccine, 3 doses of polio vaccine, and 1 dose of a measles-containing vaccine as determined by an independent state assessment of clinic coverage levels. RESULTS: Univariate analysis showed that higher coverage levels were significantly (P<.05) associated with smaller clinic size, higher proportions of clientele enrolled in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), being a nonurban clinic, and numerous vaccination practices and policies. Multivariable analysis showed that only 8 of greater than 150 factors remained associated with higher coverage levels, including having no waiting time to be seen, having telephone reminder systems, conducting home visits for defaulters, and restricting WIC vouchers when a child was undervaccinated. Motivational factors related to higher coverage included clinic lead nurses receiving an incentive to raise coverage and lead nurses participating in assessments of clinic coverage levels by state immunization staff. CONCLUSIONS: No single factor is responsible for raising vaccination coverage levels. Efforts to improve coverage should include local assessment to provide feedback on performance and identify appropriate local solutions. Coordinating with WIC, conducting recall and reminder activities, motivating clinic staff, and having staff participate in decisions are important in raising vaccination levels. PMID- 10665608 TI - Underlying causes of recurrent pneumonia in children. AB - OBJECTIVES: To determine the relative frequency of underlying factors for recurrent pneumonia and the proportion of patients in whom the underlying illness diagnosis was known prior to pneumonia recurrence. METHODS: Retrospective medical record review for a 10-year period from January 1987 through December 1997 at The Hospital for Sick Children in Toronto, Ontario, a tertiary care pediatric hospital. Recurrent pneumonia was defined as at least 2 pneumonia episodes in a 1 year period or at least 3 during a lifetime. RESULTS: Of 2952 children hospitalized with pneumonia, 238 (8%) met criteria for recurrent pneumonia. An underlying illness diagnosis was identified in 220 (92%). Of these, the underlying illness was diagnosed prior to pneumonia in 178 (81%), with the first episode in 25 (11%), and during recurrence in 17 (8%). Underlying illnesses included oropharyngeal incoordination with aspiration syndrome (114 cases [48%]), immune disorder (24 [10%]), congenital cardiac defects (22 [9%]), asthma (19 [8%]), pulmonary anomalies (18 [8%]), gastroesophageal reflux (13[5%]), and sickle cell anemia (10 [4%]). Clinical clues to diagnosis were recurrent infections at other locations and failure to thrive in the cases of an immune disorder, recurrences involving the same location in those with underlying pulmonary pathology, the association of respiratory symptoms with feeding in those with gastroesophageal reflux, or recurrent wheezing in asthmatic children. CONCLUSIONS: Recurrent pneumonia occurs in fewer than one tenth of all children hospitalized with pneumonia. Most of them have a known predisposing factor. The most common cause was oropharyngeal incoordination. PMID- 10665609 TI - Compliance with prescription filling in the pediatric emergency department. AB - OBJECTIVES: To determine the rate of compliance with filling of prescriptions written in a pediatric emergency department and to examine the reasons for not filling the prescriptions. DESIGN: Compliance with filling prescriptions was determined using a follow-up standardized telephone questionnaire, designed so that it was not obvious that assessing prescription filling was the major reason for the study. Compliance herein was defined as having the prescription filled on the same or next day of the pediatric emergency department visit. SETTING: Pediatric emergency department of a tertiary care hospital. SUBJECTS: Pediatric patients discharged home with a drug prescription. MAIN OUTCOME MEASURE: The proportion of prescriptions written in the pediatric emergency department that were filled on either the same or next day as determined by telephone follow-up. This outcome is expressed as a proportion with 95% confidence interval. RESULTS: Follow-up was completed in 1014 (83%) of the 1222 children, aged 4.5 +/- 4.2 (mean +/- SD) years. Compliance with prescription filling was 92.7% (940/1014). Parental reasons for not filling the prescription included medication unnecessary (27%), financial (6.8%), and not enough time (6.8%). Dissatisfaction with the explanation of the medical problem, instructions for treatment, and instructions for follow-up treatment were significantly associated with noncompliance by univariable logistic regression (P<.05). CONCLUSION: The rate of prescription nonfilling in children seen in a pediatric emergency department is at least 7%, although lower than that in adults in a similar setting. PMID- 10665610 TI - Radiological case of the month. Focal nodular hyperplasia of the liver. PMID- 10665611 TI - Picture of the month. Phytophotodermatitis. PMID- 10665612 TI - Pathological case of the month. Solitary intestinal fibromatosis. PMID- 10665613 TI - Choice of antibiotics in febrile neonates. PMID- 10665614 TI - Reduced prefrontal gray matter volume and reduced autonomic activity in antisocial personality disorder. AB - BACKGROUND: Major damage to gray and white matter in the prefrontal cortex and autonomic deficits have been found to result in pseudopsychopathic personality in patients with neurological disorders, but it is not known whether people with antisocial personality disorder (APD) in the community who do not have discernable brain trauma also have subtle prefrontal deficits. METHODS: Prefrontal gray and white matter volumes were assessed using structural magnetic resonance imaging in 21 community volunteers with APD (APD group) and in 2 control groups, comprising 34 healthy subjects (control group), 26 subjects with substance dependence (substance-dependent group), and 21 psychiatric controls. Autonomic activity (skin conductance and heart rate) was also assessed during a social stressor in which participants gave a videotaped speech on their faults. RESULTS: The APD group showed an 11.0% reduction in prefrontal gray matter volume in the absence of ostensible brain lesions and reduced autonomic activity during the stressor. These deficits predicted group membership independent of psychosocial risk factors. CONCLUSIONS: To our knowledge, these findings provide the first evidence for a structural brain deficit in APD. This prefrontal structural deficit may underlie the low arousal, poor fear conditioning, lack of conscience, and decision-making deficits that have been found to characterize antisocial, psychopathic behavior. PMID- 10665615 TI - Effects of supraphysiologic doses of testosterone on mood and aggression in normal men: a randomized controlled trial. AB - BACKGROUND: Field studies of illicit anabolic-androgenic steroid users suggest that some develop manic or aggressive reactions to these drugs-a potential public health problem. However, controlled laboratory evaluations of these effects remain limited. METHODS: In a randomized, placebo-controlled, crossover trial, we administered testosterone cypionate for 6 weeks in doses rising to 600 mg/wk and placebo for 6 weeks, separated by 6 weeks of no treatment, to 56 men aged 20 to 50 years. Psychiatric outcome measures included the Young Mania Rating Scale (YMRS), the Point Subtraction Aggression Paradigm (a computerized provocation test of aggression), the Aggression Questionnaire of Buss and Perry, the Symptom Checklist-90-R, daily diaries of manic and depressive symptoms, and similar weekly diaries completed by a "significant other" who knew the participant well. RESULTS: Testosterone treatment significantly increased manic scores on the YMRS (P = .002), manic scores on daily diaries (P = .003), visual analog ratings of liking the drug effect (P = .008), and aggressive responses on the Point Subtraction Aggression Paradigm (P = .03). Drug response was highly variable: of 50 participants who received 600 mg/wk of testosterone cypionate, 42 (84%) exhibited minimal psychiatric effects (maximum YMRS score, <10), 6 (12%) became mildly hypomanic (YMRS score, 10-19), and 2 (4%) became markedly hypomanic (YMRS score, > or =20). The 8 "responders" and 42 "nonresponders" did not differ significantly on baseline demographic, psychological, laboratory, or physiological measures. CONCLUSIONS: Testosterone administration, 600 mg/wk increased ratings of manic symptoms in normal men. This effect, however, was not uniform across individuals; most showed little psychological change, whereas a few developed prominent effects. The mechanism of these variable reactions remains unclear. PMID- 10665616 TI - A double-blind, placebo-controlled trial of testosterone therapy for HIV-positive men with hypogonadal symptoms. AB - BACKGROUND: The goal was to evaluate the efficacy of testosterone in alleviation of hypogonadal symptoms (diminished libido, depressed mood, low energy, and depleted muscle mass) in men with symptomatic human immunodeficiency virus illness. METHODS: Seventy-four patients were enrolled in a double-blind, placebo controlled 6-week trial with bi-weekly testosterone injections, followed by 12 weeks of open-label maintenance treatment. Major outcome measures were Clinical Global Impressions Scale ratings for libido, mood, energy, and erectile function; Hamilton Depression Rating Scale scores, and Chalder Fatigue Scale scores. Body composition changes were assessed with bioelectric impedance analysis. RESULTS: Seventy men completed the 6-week trial. Response rates, defined as much or very much improved libido, were 74% (28/38) for patients randomized to testosterone, and 19% (6/32) for placebo-treated patients (P<.001). Of the 62 completers with fatigue at baseline, 59% (20/34) receiving testosterone and 25% (7/28) receiving placebo reported improved energy (P<.01). Among the 26 completers with an Axis I depressive disorder at baseline, 58% of the testosterone-treated patients reported improved mood compared with 14% of placebo-treated patients (Fisher exact test = .08). With testosterone treatment, average increase in muscle mass over 12 weeks was 1.6 kg for the whole group, and 2.2 kg for the 14 men with wasting at baseline. Improvement on all parameters was maintained during subsequent open-label treatment for up to 18 weeks. CONCLUSION: Testosterone is well tolerated and effective in the short-term treatment of symptoms of clinical hypogonadism in men with symptomatic human immunodeficiency virus illness, restoring libido and energy, alleviating depressed mood, and increasing muscle mass. PMID- 10665617 TI - Time course of effects of testosterone administration on sexual arousal in women. AB - BACKGROUND: The assumption that testosterone is involved in human female sexual functioning is mainly based on results of studies of women with hypogonadotropic hypogonadism. This study sought to determine the effect of testosterone administration on physiological and subjective sexual arousal in sexually functional women. METHODS: In a double-masked, randomly assigned, placebo controlled crossover design, we examined whether administration of a single dose of testosterone to sexually functional women increases vaginal and subjective sexual arousal when they are exposed to erotic visual stimuli. To search for a time lag in the effect of testosterone therapy, we exposed 8 healthy women to 6 erotic film excerpts depicting intercourse. The first and second excerpts were shown immediately before and 15 minutes after, respectively, intake of placebo or testosterone; the last 4 excerpts were then shown at 1(1/2)-hour intervals. RESULTS: Sublingual intake of testosterone caused a sharp increase in plasma testosterone levels within 15 minutes; these levels declined to baseline values within 90 minutes. Three to 4(1/2) hours after reaching peak testosterone level, we found a statistically significantly increase in genital responsiveness (P = .04). Furthermore, on the day of testosterone treatment, there also was a strong and statistically significant association between the increase in genital arousal and subjective reports of "genital sensations" (P = .02) and "sexual lust" (P = .01) after 4(1/2) hours. CONCLUSIONS: There is a time lag in the effect of sublingually administered testosterone on genital arousal in women. In addition, a consecutive increase in vaginal arousal might cause higher genital sensations and sexual lust. PMID- 10665618 TI - Extended-release physostigmine in Alzheimer disease: a multicenter, double-blind, 12-week study with dose enrichment. Physostigmine Study Group. AB - BACKGROUND: The efficacy of extended-release physostigmine salicylate, an acetylcholinesterase inhibitor, was evaluated in 850 subjects with mild-to moderate Alzheimer disease (AD) in a multicenter trial. METHODS: Subjects initially entered a dose-enrichment phase in which they received 1 week each of physostigmine salicylate, 24 mg/d and 30 mg/d, and daily placebo. Among the subjects who completed this phase, 35.9% responded to physostigmine treatment, whereas 62.4% were considered nonresponders, and 1.6% could not be evaluated because of missing data. After a 4-week placebo-washout phase, 176 responder subjects were randomized to receive their best dose of physostigmine or placebo in a 12-week double-blind phase. Primary efficacy measures included the cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-Cog), the Clinician's Interview-Based Impression of Change With Caregiver Input (CIBIC+), and the Clinical Global Impression of Change (CGIC). RESULTS: In the intent-to-treat analysis of the double-blind phase, physostigmine-treated subjects scored -2.02 points better than placebo-treated subjects on the ADAS-Cog (F1,167 = 6.42 [P = .01]) and 0.33 points higher on the CIBIC+ (F1,150 = 5.68 [P = .02]). No significant improvement was observed on the CGIC or the secondary outcome measures. Nausea and vomiting were experienced by 47.0% of all physostigmine treated subjects during the double-blind phase. CONCLUSIONS: Physostigmine demonstrated a statistically significant benefit compared with placebo on a clinical global rating of change and an objective test of cognitive function. Given the frequency of gastrointestinal side effects, the role of this agent in clinical use remains to be determined. PMID- 10665619 TI - A randomized controlled trial of cognitive-behavioral therapy for persistent symptoms in schizophrenia resistant to medication. AB - BACKGROUND: Research evidence supports the efficacy of cognitive-behavioral therapy in the treatment of drug-refractory positive symptoms of schizophrenia. Although the cumulative evidence is strong, early controlled trials showed methodological limitations. METHODS: A randomized controlled design was used to compare the efficacy of manualized cognitive-behavioral therapy developed particularly for schizophrenia with that of a nonspecific befriending control intervention. Both interventions were delivered by 2 experienced nurses who received regular supervision. Patients were assessed by blind raters at baseline, after treatment (lasting up to 9 months), and at a 9-month follow-up evaluation. Patients continued to receive routine care throughout the study. An assessor blind to the patients' treatment groups rated the technical quality of audiotaped sessions chosen at random. Analysis was by intention to treat. RESULTS: Ninety patients received a mean of 19 individual treatment sessions over 9 months, with no significant between-group differences in treatment duration. Both interventions resulted in significant reductions in positive and negative symptoms and depression. At the 9-month follow-up evaluation, patients who had received cognitive therapy continued to improve, while those in the befriending group did not. These results were not attributable to changes in prescribed medication. CONCLUSION: Cognitive-behavioral therapy is effective in treating negative as well as positive symptoms in schizophrenia resistant to standard antipsychotic drugs, with its efficacy sustained over 9 months of follow-up. PMID- 10665620 TI - Brain serotonin1A receptor binding measured by positron emission tomography with [11C]WAY-100635: effects of depression and antidepressant treatment. AB - BACKGROUND: Pharmacological and postmortem investigations suggest that patients with major depressive disorder have alterations in function or density of brain serotonin1A (5-HT1A) receptors. The aim of the present study was to use positron emission tomography with the selective 5-HT1A receptor antagonist [11C]WAY-100635 to measure 5-HT1A receptor binding in depressed patients before and during treatment with selective serotonin reuptake inhibitors. METHODS: Positron emission tomographic scans with [11C]WAY-100635 were performed on 25 patients with major depressive disorder. These included 15 unmedicated depressed patients. Ten of these unmedicated patients were scanned again during selective serotonin reuptake inhibitor treatment. A further 10 patients with major depressive disorder were scanned on one occasion only while taking selective serotonin reuptake inhibitors. Comparisons were made with [11C]WAY-100635 positron emission tomographic scans in 18 healthy volunteer subjects. Region of interest analysis and statistical parametric mapping were performed on binding potential images generated using a reference tissue model. RESULTS: Binding potential values were reduced across many of the regions examined, including frontal, temporal, and limbic cortex in both unmedicated and medicated depressed patients compared with healthy volunteers. Binding potential values in medicated patients were similar to those in unmedicated patients. CONCLUSIONS: Major depressive disorder is associated with a widespread reduction in 5-HT1A receptor binding. This reduced 5 HT1A receptor binding was not changed by selective serotonin reuptake inhibitor treatment. PMID- 10665621 TI - Neurologic soft signs in chronic posttraumatic stress disorder. AB - BACKGROUND: Subtle neurologic impairment has been reported in several mental disorders. The goals of the present study were to evaluate neurologic status in patients of both sexes with chronic posttraumatic stress disorder (PTSD) from different traumatic experiences. METHODS: Twenty-one adult women who were sexually abused as children (12 with PTSD, 9 without) and 38 male Vietnam War combat veterans (23 with PTSD, 15 without) underwent examination for 41 neurologic soft signs, which were scored by the examiner as well as a blind rater observing videotapes. Subject history was obtained with special attention to neurodevelopmental problems. Psychometrics included the Wender Utah Rating Scale for symptoms of childhood attention-deficit/hyperactivity disorder and the Michigan Alcoholism Screening Test. Veterans also completed the Combat Exposure Scale and subtests of the Wechsler Adult Intelligence Scale-Revised. RESULTS: Average neurologic soft sign scores (interrater reliability = 0.74) of women with PTSD owing to sexual abuse in childhood (mean [SD], 0.77 [0.32]) and veteran men (0.72 [0.20]) with combat-related PTSD were comparable and significantly (P<.001) higher than those of women sexually abused as children (0.42 [0.10]) and combat veteran men (0.43 [0.17]) without PTSD. This effect could not be explained by a history of alcoholism or head injury. Subjects with PTSD reported more neurodevelopmental problems and more childhood attention-deficit/hyperactivity disorder symptoms and had lower IQs, all of which were significantly correlated with neurologic soft signs. CONCLUSION: Neurologic compromise is evident from subject history and findings from physical examination in both women and men with chronic PTSD who had experienced different kinds of traumatic events in childhood and adulthood. PMID- 10665622 TI - Does lithium treatment still work? Evidence of stable responses over three decades. AB - To evaluate whether lithium treatment has been overvalued and may be no longer as effective as formerly, we reviewed published reports on long-term lithium treatment (1970-1996) as well as analyzing its clinical effects on 360 patients with DSM-IV bipolar disorder who entered into lithium maintenance monotherapy after 1970. Neither reported recurrence rates nor average proportions of time ill nor patient improvement of 50% or more during lithium maintenance therapy in a stable clinic setting has changed significantly since the 1970s. Unfavorable results in some settings may reflect accumulation over time of patients with complex, less treatment-responsive illnesses. Lithium is unmatched in research support for long-term clinical effectiveness against morbidity and mortality associated with depression or mania in bipolar I and II disorders. Data evaluated herein did not support suggestions that benefits of lithium have been exaggerated in the past or have been lost recently. PMID- 10665623 TI - Discrepancies in the efficacy of lithium. PMID- 10665624 TI - Social phobia or social anxiety disorder: what's in a name? PMID- 10665625 TI - Molecular neurobiology for practicing psychiatrists, part 4: transferring the message of chemical neurotransmission from presynaptic neurotransmitter to postsynaptic gene expression. AB - Neurotransmitters activate genes in their target neurons by precipitating a molecular cascade, which may be the ultimate consequence of chemical neurotransmission. When this transfer is aberrant, a mental disorder may be manifest. When drugs act upon neurons to change gene expression, this could lead to therapeutic actions, side effects, and the long-term consequences of drug abuse. PMID- 10665626 TI - Safety and tolerability of oral loading divalproex sodium in acutely manic bipolar patients. AB - BACKGROUND: Achieving therapeutic blood levels of a mood stabilizer as quickly as possible is desirable in patients with acute mania. We examined the feasibility and safety of an accelerated oral loading strategy (divalproex, 30 mg/kg/day, on days 1 and 2, followed by 20 mg/kg/day on days 3-10) designed to bring serum valproate concentrations to therapeutic levels (i.e., above 50 microg/mL). METHOD: Fifty-nine patients who met DSM-IV diagnostic criteria for current manic episode and who had a Mania Rating Scale score > or = 14 were randomly assigned on a double-blind basis to receive divalproex oral loading (N = 20); divalproex nonloading (N = 20) at a starting dose of 250 mg t.i.d. on days 1 and 2, followed by standard dose titration for days 3 to 10; or lithium carbonate (N = 19) at a starting dose of 300 mg t.i.d., followed by standard dose titration for days 3 to 10. RESULTS: Eighty-four percent of the divalproex-loading patients, but only 30% of the divalproex-nonloading patients, had valproate serum levels above 50 microg/mL at day 3 of the study. None of the lithium-treated patients had serum lithium levels above 0.8 mEq/L at study day 3. No patient was removed from the study because of an adverse event. There were no significant differences between the groups in the frequencies or types of adverse events. CONCLUSION: Accelerated oral loading with divalproex sodium is a feasible and safe method to bring serum valproate concentrations to effective levels rapidly. PMID- 10665627 TI - The emergence of social phobia during clozapine treatment and its response to fluoxetine augmentation. AB - BACKGROUND: The underlying neurochemical basis of social phobia has yet to be fully explained, but there are suggestions of serotonergic and dopaminergic dysfunction. The atypical neuroleptic clozapine has been reported to induce anxiety symptoms, probably owing to its effect on serotonergic pathways. We report 12 cases of schizophrenic patients who developed social phobia during clozapine treatment. METHOD: Patients were assessed using the Structured Clinical Interview for DSM-III-R, Patient Version, Scale for the Assessment of Negative Symptoms, Scale for the Assessment of Positive Symptoms, the Liebowitz Social Phobia Scale, and the Brief Psychiatric Rating Scale. They were reevaluated after 12 weeks of cotreatment with clozapine and fluoxetine. RESULTS: In 8 of the 12 cases, symptoms responded (> or = 35% reduction in Liebowitz Social Phobia Scale score) with an adjunctive regimen of fluoxetine. CONCLUSION: Data are discussed in light of neurochemical mechanisms and cognitive adaptations that could explain the onset of anxiety spectrum disorders (such as social phobia) in clozapine treated schizophrenic subjects during remission of psychotic symptoms. PMID- 10665628 TI - Multicenter, placebo-controlled, fixed-dose study of citalopram in moderate-to severe depression. AB - BACKGROUND: Citalopram, the most selective serotonin reuptake inhibitor (SSRI), is a bicyclic phthalane derivative with a chemical structure that is unrelated to that of other SSRIs and available antidepressants. The drug is approved for use in 69 countries. This 6-week, fixed-dose, placebo-controlled, parallel-arm, multicenter trial was performed to confirm its efficacy and safety in treatment of outpatients with major depression in the United States. METHOD: Six hundred and fifty adult outpatients with moderate-to-severe major depression (DSM-III-R) were randomly assigned to receive citalopram at doses of 10 mg (N = 131), 20 mg (N = 130), 40 mg (N = 131), or 60 mg (N = 129) or placebo (N = 129) once daily. Outcome assessments were the 21-item Hamilton Rating Scale for Depression (HAM D), the Montgomery-Asberg Depression Rating Scale (MADRS), and the Clinical Global Impressions scale. RESULTS: Between-group comparisons of the change from baseline to endpoint revealed significantly greater improvement in the citalopram patients relative to the placebo patients on all 3 efficacy measures. Patients randomly assigned to 40 mg/day and 60 mg/day of citalopram showed significantly greater improvement than placebo on all efficacy measures, as well as on the HAM D symptom clusters measuring depressed mood, melancholia, cognitive disturbance, and psychomotor retardation. Patients who received 10 mg/day and 20 mg/day of citalopram also showed consistent improvement relative to placebo on all efficacy ratings, with statistical significance demonstrated in the MADRS response rate, the HAM-D depressed mood item, and the HAM-D melancholia subscale. Citalopram was well tolerated, with only 15% of patients discontinuing for adverse events. The side effects most commonly associated with citalopram treatment were nausea, dry mouth, somnolence, insomnia, and increased sweating. CONCLUSION: Citalopram was significantly more effective than placebo in the treatment of moderate-to-severe major depression, especially symptoms of depressed mood and melancholia, with particularly robust effects shown at doses of 40 and 60 mg/day. Citalopram was well tolerated in spite of forced upward titration to fixed-dose levels, with a low incidence of anxiety, agitation, and nervousness. PMID- 10665629 TI - Paroxetine, clomipramine, and cognitive therapy in the treatment of panic disorder. AB - BACKGROUND: This 12-week, placebo-controlled study was carried out to compare the relative efficacy of paroxetine, clomipramine, and cognitive therapy in the treatment of DSM-III-R-defined panic disorder with or without agoraphobia. METHOD: After a 3-week single-blind, placebo run-in period, 131 patients were randomly assigned to receive double-blind medication or 12 sessions of cognitive therapy based on the model of Clark. Efficacy assessments included the daily panic attack diary, the Clinical Global Impression scale, the Patient Global Evaluation, the Hamilton Rating Scale for Anxiety, the Marks-Sheehan Phobia Scale, the Montgomery-Asberg Depression Rating Scale, and the Sheehan Disability Scale. RESULTS: Comparisons with placebo revealed significant superiority of paroxetine (20-60 mg/day) and clomipramine (50-150 mg/day) on nearly all outcome measures. On most measures, paroxetine also showed higher efficacy than cognitive therapy. With few exceptions, cognitive therapy did not differ significantly from placebo. The number of subjects becoming panic-free (66%) was higher and the onset of action was faster in the paroxetine-treated group. Treatment with cognitive therapy yielded the highest drop-out rate (26%). CONCLUSION: In this short-term study assessing treatment of panic disorder and agoraphobia, paroxetine and clomipramine were consistently superior to pill placebo, whereas cognitive therapy was superior on only a few measures. PMID- 10665630 TI - Clinical features, psychiatric comorbidity, and health-related quality of life in persons reporting compulsive computer use behavior. AB - BACKGROUND: We sought to examine the demographic and clinical features and psychiatric comorbidity in persons reporting compulsive computer use. METHOD: Sixteen men and 5 women were recruited by advertisement and word-of-mouth. All reported excessive computer use that interfered with social or occupational functioning or caused personal distress. The subjects completed structured and semistructured assessments, including a computer version of the Diagnostic Interview Schedule (DIS), the Minnesota Impulsive Disorders Interview, the Personality Diagnostic Questionnaire-Revised (PDQ-R), and a brief version of the Medical Outcome Study Short Form-36 (SF-36). RESULTS: The typical subject was a 32-year-old single white man with a mean yearly income of $27,000; problem computer use began at age 29 and consumed 27 hours each week. Eleven subjects (52%) reported school or academic problems resulting from their computer use, and 12 (57%) reported that family members had confronted them about it. Thirteen subjects (62%) had tried to cut back on their computer usage. Nine subjects (43%) reported missing work or school owing to their computer use. According to DIS results, 7 subjects (33%) had a lifetime mood disorder, 8 subjects (38%) had a substance use disorder, and 4 subjects (19%) had a lifetime anxiety disorder. According to the PDQ-R results, 11 subjects (52%) met criteria for at least one personality disorder, the most frequent being the borderline, antisocial, and narcissistic types. Impulse-control disorders were also common, particularly compulsive buying. On the SF-36, subjects showed impaired mental health functioning compared with a normative sample. CONCLUSION: The results show that persons reporting compulsive computer use suffer substantial psychiatric comorbidity and show evidence of emotional distress. While the disorder appears to be increasing in prevalence, more work is needed to determine its relationship with other disorders and to determine its risk factors, family history, psychosocial complications, and natural history. PMID- 10665631 TI - Venlafaxine in the treatment of dysthymia: an open-label study. AB - BACKGROUND: Numerous studies have demonstrated the effectiveness of antidepressant medications in the treatment of dysthymia, or chronic mild depression. Venlafaxine blocks reuptake of both serotonin and norepinephrine and may produce a more complete antidepressant response than do single-mechanism selective serotonin reuptake inhibitors. The purpose of this open-label study was to provide preliminary data on the tolerability and effectiveness of venlafaxine for patients with dysthymia. METHOD: Twenty-two dysthymic subjects (DSM-III-R criteria) were enrolled in this 10-week, open-label trial, and 5 dropped out prior to their second visit. Seventeen subjects (77.3%) received more than 1 week of medication. RESULTS: Of these 17 subjects, 13 (76.5%) were treatment responders. Results of paired sample t tests were highly significant, indicating that, on average, there was significant improvement on all measures of symptomatology and functioning, with mean +/- SD scores on the Hamilton Rating Scale for Depression decreasing from 20.95 +/- 6.50 at baseline to 6.06 +/- 5.49 at week 10. The mean +/- SD final dose was 178.68 +/- 70.80 mg/day. Side effects were reported by 17 (85%) of the 20 subjects for whom tolerability was assessed (the most common were fatigue, dry mouth, and nausea); 5 (22.7%) of 22 patients discontinued treatment because of side effects, primarily nausea (N = 3). CONCLUSION: These findings suggest the benefit of venlafaxine in the treatment of chronic depression and the need for more rigorous studies. PMID- 10665632 TI - Effectiveness of antipsychotic therapy in a naturalistic setting: a comparison between risperidone, perphenazine, and haloperidol. AB - BACKGROUND: Therapeutic ineffectiveness and noncompliance with antipsychotic agents are major contributors to rehospitalization in patients with psychotic disorders. It is unknown whether risperidone's favorable side effect profile compared with that of the conventional antipsychotics results in improved compliance and reduced hospitalizations in a naturalistic setting. The purpose of this study was to test the hypothesis that treatment with risperidone reduces readmission rates and associated costs when compared with treatment with perphenazine or haloperidol. METHOD: Inpatients prescribed either risperidone, perphenazine, or haloperidol between January 1, 1995, and December 31, 1995, as a single oral antipsychotic at discharge were retrospectively identified. Data were collected for that index hospitalization and for a 1-year follow-up period. Primary outcome measures included re-admission rates, changes in antipsychotic therapy, anticholinergic drug use, and costs. RESULTS: There were 202 evaluable patients (81 treated with risperidone, 78 with perphenazine, and 43 with haloperidol). Baseline demographics were similar between groups except that more patients in the risperidone group had a primary diagnosis of psychotic disorder or had been hospitalized in the year prior to study. The percentage of patients readmitted during the 1-year follow-up period was similar among drug groups (41% risperidone, 26% perphenazine, and 35% haloperidol) when controlled for baseline differences in diagnosis and hospitalization history (p = .32). Anticholinergic drug use was more common in the haloperidol group (p = .004). Mean yearly cost (drug + hospitalization) in the risperidone group was $20,317, nearly double that in the other treatment groups (p < .001). CONCLUSION: The results from this naturalistic study indicate that the high cost of risperidone is not offset by a reduction in readmission rates when compared with conventional antipsychotics. PMID- 10665633 TI - Buspirone as an antidote to SSRI-induced bruxism in 4 cases. AB - BACKGROUND: One hypothesis to explain selective serotonin reuptake inhibitor (SSRI)-induced bruxism states that SSRIs increase extrapyramidal serotonin levels, thereby inhibiting dopaminergic pathways controlling movement. Previous reports have emphasized buspirone's postsynaptic dopaminergic effect as a partial antidote to the suppressed dopamine levels. CASE REPORTS: Four patients, recently started on treatment with the SSRI sertraline, presented with new-onset complaints attributable to SSRI-induced bruxism. All 4 responded to adjunctive buspirone, a serotonin-1A (5-HT1A) receptor agonist, with relief of bruxism and associated symptoms. DISCUSSION: We expand the hypothesis put forth in previous reports by proposing that buspirone is not only acting postsynaptically in the extrapyramidal system, but also presynaptically on serotonergic neurons that influence masticatory modulation in the mesocortical tract. Our 4 cases support the concept of buspirone acting as a full agonist at the presynaptic 5-HT1A somatodendritic receptors located on the cell bodies of raphe serotonergic neurons that project to the ventral tegmental area (VTA) of the midbrain. These serotonergic neurons modulate the firing of the mesocortical tract, which itself projects from the VTA to the prefrontal cortex and acts on masticatory muscle activity through inhibiting spontaneous movements such as bruxism. While the literature is confusing and contradictory on definitions of bruxism and etiologies of incompletely understood movement disorders, we believe SSRI-induced bruxism is best conceptualized as a form of akathisia. PMID- 10665634 TI - Two-year maintenance treatment with citalopram, 20 mg, in unipolar subjects with high recurrence rate. AB - BACKGROUND: The efficacy of citalopram, 20 to 60 mg/day, in relapse prevention in major depression was demonstrated in 6-month placebo-controlled studies. The authors tested the efficacy of citalopram, 40 mg/day, in relapse prevention over a 4-month period and citalopram, 20 mg/day, in recurrence prevention over a 24 month period. METHOD: Fifty inpatients with recurrent major depressive disorder (DSM-IV criteria) who had had at least one depressive episode during the 18 months preceding the index episode were openly treated with citalopram, 40 mg/day. Thirty-six subjects had a stable response to citalopram and remained in the continuation treatment with citalopram, 40 mg/day, for 4 months as outpatients. At the time of recovery, 32 patients gave their written informed consent before entering the 24-month maintenance period with citalopram, 20 mg/day. They were evaluated monthly by trained psychiatrists on the basis of the 21-item Hamilton Rating Scale for Depression. Every 3 months, patients were given the Sheehan Disability Scale, a self-rating instrument, to assess their psychosocial adjustment. RESULTS: No relapse was observed in the 4-month continuation period. Sixteen (50%) of 32 patients who entered the 24-month maintenance period had a new recurrence. Patients with recurrence showed a persistent moderate disability on Sheehan Disability Scale score, while no further differences were highlighted in clinical and demographic characteristics between patients with and without recurrence. CONCLUSION: In agreement with previous findings, these data suggest that a full dose of antidepressant is strongly recommended in prophylactic therapy of patients with recurrent major depression. Moreover, it appears that psychosocial impairment may increase the risk of recurrence, thus conditioning a poor outcome. PMID- 10665635 TI - Sildenafil treatment of antidepressant-induced sexual dysfunction. PMID- 10665636 TI - Improved outcome in fluvoxamine-treated patients with SSRI-induced sexual dysfunction. PMID- 10665637 TI - Choreiform dyskinesia with acute onset and protracted course following fluoxetine treatment. PMID- 10665638 TI - Deliberate self-poisoning following fluvoxamine-neuroleptics combination. PMID- 10665639 TI - Penile anesthesia associated with sertraline use. PMID- 10665640 TI - Tardive dyskinesia associated with olanzapine monotherapy. PMID- 10665641 TI - Schizoaffective disorder: a form of schizophrenia or affective disorder? AB - BACKGROUND: The diagnostic status of schizoaffective disorder continues to be controversial. Researchers have proposed that schizoaffective disorder represents a variant of schizophrenia or affective disorder, a combination of the 2, or an intermediate condition along a continuum between schizophrenia and affective disorder. METHOD: We compared outpatients aged 45 to 77 years with DSM-III-R diagnosis of schizoaffective disorder (N = 29), schizophrenia (N = 154), or nonpsychotic mood disorder (N = 27) on standardized rating scales of psychopathology and a comprehensive neuropsychological test battery. A discriminant function analysis was used to classify the schizoaffective patients based on their neuropsychological profiles as being similar either to schizophrenia patients or to those with nonpsychotic mood disorder. RESULTS: The schizoaffective and schizophrenia patients had more severe dyskinesia, had a weaker family history of mood disorder, had been hospitalized for psychiatric reasons more frequently, were more likely to be prescribed neuroleptic and anticholinergic medication, and had somewhat less severe depressive symptoms than the mood disorder patients. The schizophrenia patients had more severe positive symptoms than the schizoaffective and mood disorder patients. The neuropsychological performances of the 2 psychosis groups were more impaired than those of the nonpsychotic mood disorder patients. Finally, on the basis of a discriminant function analysis, the schizoaffective patients were more likely to be classified as having schizophrenia than a mood disorder. CONCLUSION: These findings suggest that schizoaffective disorder may represent a variant of schizophrenia in clinical symptom profiles and cognitive impairment. PMID- 10665642 TI - Beta-catenin accumulation and mutation of exon 3 of the beta-catenin gene in hepatocellular carcinoma. PMID- 10665643 TI - Reduced T cell response in carcinogen-sensitive Donryu rats compared with carcinogen-resistant DRH rats. AB - Carcinogen-resistant DRH rats were developed from carcinogen-sensitive Donryu rats, which showed a high incidence of hepatic tumors when they were exposed to 3'-methyl-4-dimethyl-amino-azobenzene (3'-MeDAB4) or other aminoazo hepatocarcinogens. In order to study the mechanism of the difference of carcinogenesis, we studied the immunological competence of Donryu rats compared with that of DRH rats. Anti-keyhole limpet hemocyanin (KLH) antibody and KLH specific delayed hypersensitivity (DTH) responses after immunization with KLH were reduced in Donryu rats compared with DRH rats. Proliferative responses of spleen cells to KLH and nonspecific mitogens such as conconavalin A (Con A) and phytohemagglutinin (PHA) were significantly lower in Donryu rats than in DRH rats. Upon the cross-linking of T cell receptor (TCR) complex using anti-CD3 monoclonal antibody (Mab), spleen cells from Donryu rats proliferated poorly. Two other strains of rats, SD and Wistar, exhibited high responsiveness, comparable to that of DRH rats, indicating that the responsiveness of Donryu rats was impaired. The production of interleukin-2 (IL-2) upon stimulation with Con A and the responsiveness of Con A blasts to exogenous IL-2 were also attenuated in Donryu rats. In contrast to T cell responsiveness, natural killer (NK) cell activity of spleen was increased in Donryu rats. Flow cytometric analysis revealed that the expression of CD4 and CD8 on T cells was decreased in Donryu rats, though the expression of other T cell markers such as CD2, CD3 and CD5 was not different. These results indicate that Donryu rats, which have been used in many years for cancer research in Japan, have impaired immunological surveillance mechanisms. This is likely to be one of the factors accounting for the high sensitivity to chemical carcinogens and the high susceptibility to transplanted tumor cells of Donryu rats. PMID- 10665644 TI - Epstein-Barr virus-encoded latent membrane protein 1 co-expresses with epidermal growth factor receptor in nasopharyngeal carcinoma. AB - Latent membrane protein 1 (LMP-1) is the only Epstein-Barr virus (EBV)-encoded oncogenic protein that has been detected in nasopharyngeal carcinoma (NPC), a cancer that is closely associated with EBV. Previous in-vitro studies have demonstrated that LMP-1 can upregulate epidermal growth factor receptor (EGFR) in epithelial cells. It was not established whether this cellular effect exists in NPC. To assess the association between LMP-1 and EGFR in NPC tissues, 60 NPC specimens were examined by immunohistochemistry using anti-LMP-1 antibody (CS 1 4) and anti-EGFR antibodies (EGFR 1, EGFR 1005). The results revealed that 41 (68.3%) specimens were immunopositive for LMP-1 and 44 (73.3%) specimens over expressed EGFR. Morphologically, the expressions of LMP-1 and EGFR were homogeneously distributed in the tumor nests. In addition, the correlation between LMP-1 and EGFR was statistically significant (P<0.001, chi2 test, d.f. = 1). To elucidate further the correlation between LMP-1 and EGFR in vivo and in situ, an indirect dual immunofluorescence assay was conducted, using secondary antibodies conjugated with fluorescein isothiocyanate (FITC) or indocarbocyanine (Cy3). The results disclosed an intimate co-expression of LMP-1 and EGFR. In summary, the data indicate that over-expression of EGFR is a common phenomenon in NPC, and that EGFR is co-expressed with LMP-1 in NPC. Thus, EBV may play a role in the tumorigenesis of NPC through the effects of LMP-1 and EGFR. PMID- 10665645 TI - Correlation of clinicopathologic features of resected hepatocellular carcinoma with hepatitis C virus genotype. AB - Clinicopathologic findings in patients with hepatocellular carcinoma complicating hepatitis C virus and outcomes after liver resection were compared between different viral genotypes. One hundred and forty-seven patients with both anti hepatitis C virus antibody and hepatitis C virus RNA in their sera underwent curative resection for hepatocellular carcinoma in our department between 1991 and 1997. Of these patients, 115 were infected with hepatitis C virus genotype 1b (group 1), and 32 were infected with 2a or 2b (group 2). Clinicopathologic findings and outcomes after operation were compared between the two groups. Alanine aminotransferase activity was significantly higher in group 2 than in group 1. Genotypes did not differ concomitantly with histopathologic features of the carcinoma or adjacent hepatic tissue. Although the tumor-free survival rate did not differ significantly between the two groups, recurrence was not detected during the period beyond 3 years following operation in group 2, while recurrences arose during that period in 16 group 1 patients, most of whom continued to manifest active hepatitis. In 7 of these 16 patients, the recurrent tumors were histologically multicentric in origin. The cumulative survival rate was significantly lower in group 1 than 2. Multivariate analysis indicated that genotype 1b was an independent risk factor for short survival. Patients infected with genotype 1b may have a relatively high risk of ongoing hepatocarcinogenesis and more aggressive progression of associated liver dysfunction, resulting in a poorer outcome than with other genotypes. PMID- 10665646 TI - Beta-catenin accumulation and mutation of exon 3 of the beta-catenin gene in hepatocellular carcinoma. AB - A study was conducted to clarify the contribution of beta-catenin accumulation and mutation of the beta-catenin gene to hepatocarcinogenesis. Beta-catenin accumulation was examined immunohistochemically in 38 paired samples of hepatocellular carcinoma (HCC) and corresponding non-cancerous liver tissue. Gene mutation was analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and direct sequencing using intronic primers encompassing exon 3. Neither accumulation nor mutation was detected in non-cancerous liver tissues that showed no remarkable histological features, chronic hepatitis or liver cirrhosis. Accumulation of beta-catenin was seen in the nucleus, cytoplasm or cell membrane in 15 of 38 (39%) HCC samples, and gene mutation was seen in 9 of 38 (24%) HCC samples. Although there was a significant correlation between accumulation and mutation (P<0.01), six HCCs without mutation also showed accumulation. Samples of early HCC showed neither accumulation nor mutation, and accumulation and mutation were each correlated significantly with portal vein tumor involvement (P<0.05). The present results indicate that (1) mutation of exon 3 of the beta-catenin gene can lead to beta-catenin accumulation, although other mechanisms of accumulation may also operate in HCC, and (2) beta-catenin accumulation and mutation of the beta-catenin gene are not early events in hepatocarcinogenesis, and may be associated with the malignant progression of HCC. PMID- 10665647 TI - Frameshift mutations and a length polymorphism in the hMSH3 gene and the spectrum of microsatellite instability in sporadic colon cancer. AB - Mutations in the hMVSH3 gene in sporadic colon cancer with microsatellite instability (MSI) were investigated, since several mismatch repair genes were known to be mutated in cancers with MSI, but only deletions in the (A)8 region in the hMSH3 gene have been reported. We also analyzed the relationships between hMSH3 mutations and the spectrum of MSI. We screened MSI in 79 sporadic colon cancer samples using mono- and dinucleotide repeat markers and the samples with MSI were further analyzed for tri- and tetranucleotide repeat instability and mutations in the hMSH3 gene by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. Five (6%) out of 79 tumors were MSI-H and 15 (19%) were MSI-L. Two MSI-H tumors showed insertion in the (C)8 region in the hMSH6 gene and one tumor showed insertion and deletion in the (A)8 region in the hMSH3 gene, and two of the three above tumors showed MSI in tri-and tetranucleotide repeats. One MSI-L tumor showed somatic alteration in a 9-bp repeat sequence in hMSH3. No frameshift mutations were found in the (A)7 and (A)6 regions in hMSH3. Thus, we confirmed that the (A)8 region in hMSH3 is a hot spot and mutations in the (A)7 and (A)6 regions in hMSH3 are not common. The hMSH3 mutation may enhance genomic instability in some colorectal cancers. PMID- 10665648 TI - Infrequent BCL10 mutations in B-cell non-Hodgkin's lymphomas. AB - The BCL10 gene was recently isolated from the breakpoint region of t(1;14)(p22;q32) in mucosa-associated lymphoid tissue (MALT) lymphomas. Somatic mutations of BCL10 were found in not only t(1;14)-bearing MALT lymphomas, but also a wide range of other tumors. To clarify the actual frequency and spectrum of BCL10 mutations in primary B-cell non-Hodgkin's lymphomas (NHL), we examined a total of 139 NHL cases comprising 25 with MALT lymphomas, 54 with follicular B cell lymphomas (FCL), and 60 with diffuse large B-cell lymphomas (DLBL). Polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing analyses led to the identification of four nucleotide changes in FCL and one in DLBL. In contrast, no BCL10 mutations were found in our series of MALT lymphomas. While screening for mutations, we also found three polymorphic sequence variants at codons 5 and 213 and in intron 1 of the BCL10 gene. Our results strongly suggest that somatic mutations of BCL10, if they occur at all, are rare in B-cell NHLs and do not commonly contribute to their molecular pathogenesis. PMID- 10665649 TI - C-kit gene abnormalities in gastrointestinal stromal tumors (tumors of interstitial cells of Cajal. AB - Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the GI tract, and expresses KIT and CD34 in most cases. Gain-of-function mutation of the c-kit proto-oncogene has been described, but its significance in GIST has not yet been fully evaluated. Mutation in exon 11 of the c-kit gene was determined by both polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis and direct sequencing in primary and metastatic GISTs and esophageal leiomyomas in Japanese subjects. C-kit gene mutation was identified in 15 of 48 primary GISTs (31%), four of seven metastatic GISTs, but none of the leiomyomas. Three mutations were mis-sense point mutations, and 16 were in-frame deletions of 3-48 bp. C-kit gene mutation was observed equally in low- and high-risk groups, and was not related to any clinical and pathologic factors, phenotypes or Ki-67 labeling index (LI) of tumor cells. In five of 15 deletion mutations (four in primary tumors and one in a metastatic tumor), the mutations were present at the distal location of exon 11 of the c-kit gene, which was a minor mutation in previous reports from Finland and the USA. C-kit gene mutations in GIST are not always related to a poor prognosis, but further comparative studies are necessary in Western and Japanese populations. PMID- 10665650 TI - Frequent allelic imbalance on chromosome 18q21 in early superficial colorectal cancers. AB - Genetic alterations in early superficial colorectal cancers have rarely been reported. In the present study, we searched for alterations in the APC and p53 genes in 27 superficial (20 depressed and 7 elevated) and 21 protruding colorectal cancers with submucosal invasion by means of PCR-single strand conformation polymorphism. Allelic imbalance (AI) on five loci, i.e., 1p34-36, 8p21-22, 14q32, 18q21 and 22q12-13, was also analyzed. Since a high incidence of 18q21 AI was detected in the superficial depressed cases, we further screened for alterations in Smad2, Smad4 and DCC. APC alterations were observed in three superficial depressed, one superficial elevated, and 11 protruding colorectal cancers, indicating that the frequency of APC alterations in superficial depressed cases was significantly lower than that in the protruding ones. There was no significant association between p53 alterations and macroscopic types. AI on 18q21 (13/20, 65%) was much higher than those on the other four loci in the superficial depressed cases. Moreover, the frequency of 18q21 AI in the superficial depressed cases was significantly higher than that in the protruding ones. Smad4 alterations were only detected in 1 of the 13 superficial depressed and 3 of the 17 protruding cases, while Smad2 and DCC alterations were not detected in any case examined. These data suggest that the carcinogenetic pathways of protruding and superficial depressed colorectal cancers are different, and that alterations of tumor suppressor gene(s) located on 18q21 other than Smad2, Smad4 and DCC might be associated with most superficial depressed colorectal cancers. PMID- 10665651 TI - Cyclooxygenase-2 (COX-2) mRNA expression levels in normal lung tissues and non small cell lung cancers. AB - One of the cyclooxygenase (COX) isoforms, COX-2, is overexpressed in various human cancers. In this study, we examined the gene expression levels of COX-2 in primary non-small cell lung cancers (NSCLC), metastatic lymph nodes, and normal lung tissues. The expression levels of the COX-2 gene were assessed by means of the reverse transcription polymerase chain reaction in 76 autopsy samples (29 primary NSCLC, 29 corresponding normal lung tissues, and 9 metastatic lymph nodes). The expression levels in NSCLC (both adenocarcinomas and squamous cell carcinomas) were significantly higher than in normal lung tissues and were significantly higher in adenocarcinomas than in squamous cell carcinomas. Differences between the levels of expression of COX-2 in primary tumors and their corresponding metastatic lymph nodes in 9 adenocarcinoma patients were not significant. Our results indicate that COX-2 may be associated with carcinogenesis of NSCLC, and that it may be a target for the treatment of NSCLC. PMID- 10665652 TI - Expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase in human bladder cancer. AB - We investigated the expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) in primary bladder cancer, its association with clinicopathologic findings, and their prognostic value. mRNA was extracted from 20 bladder cancer specimens and 6 normal bladder mucosal tissues. Relative amounts of PD-ECGF/TP mRNA were evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) and compared with the level of glyceraldehyde 3-phosphate dehydrogenase mRNA (used as an internal standard). PD-ECGF/TP expression was examined by immunohistochemistry in 85 patients who underwent cystectomy for bladder cancer. Serum PD-ECGF/TP levels were measured in 23 patients using a sandwich-type enzyme-linked immunosorbent assay. By RT-PCR analysis, expression of PD-ECGF/TP was found to be 7-fold higher in invasive tumors than in superficial tumors (P<0.01) and 9-fold higher than in normal bladder (P<0.01). Out of 85 transitional cell carcinoma tissue samples, 69 (81%) were evaluated as PD-ECGF/TP-positive by immunohistochemical staining. PD-ECGF/TP expression correlated significantly with tumor grade (P = 0.001), depth of invasion (P = 0.012), and lymphatic invasion (P = 0.01). No correlation was found between expression of PD-ECGF/TP and the number of tumors, tumor configuration, lymph node involvement, venous invasion, c-erbB-2 expression, or overall survival. We could not detect a significant serum level of PD-ECGF/TP in any patient. The results suggest that PD-ECGF/TP might give valuable information for bladder cancer management, though it may not be a good new tumor marker for bladder cancer. PMID- 10665653 TI - Gamma-irradiation deregulates cell cycle control and apoptosis in nevoid basal cell carcinoma syndrome-derived cells. AB - The nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder characterized by nevi, palmar and plantar pits, falx calcification, vertebrate anomalies and basal cell carcinomas. It is well known in NBCCS that gamma-irradiation to the skin induces basal cell carcinomas or causes an enlargement of the tumor size, although the details of the mechanism remain unknown. We have established lymphoblastoid cell lines from three NBCCS patients, and we present here the first evidence of abnormal cell cycle and apoptosis regulations. A novel mutation (single nucleotide deletion) in the coding region of the human patched gene, PTCH, was identified in two sibling patients, but no apparent abnormalities were detected in the gene of the remaining patient. Nevertheless, the three established cell lines showed similar features in the following analyses. Flow cytometric analyses revealed that the NBCCS-derived cells were accumulated in the G2M phase after gamma-irradiation, whereas normal cells showed cell cycle arrest both in the G0G1 and G2M phases. The fraction of apoptotic cells after gamma-irradiation was smaller in the NBCCS cells. The level of p27 expression markedly decreased after gamma-irradiation in the NBCCS cells, although the effects of the irradiation on the expression profiles for p53, p21 and Rb did not differ in normal and NBCCS cells. These findings may provide a clue to the molecular mechanisms of tumorigenesis in NBCCS. PMID- 10665654 TI - Significance of heterogeneous nuclear ribonucleoprotein B1 as a new early detection marker for oral squamous cell carcinoma. AB - The development of an early tumor detection marker for oral cancer is an obvious need due to the high recurrence rate and poor survival rate. Based on our previous report that overexpression of heterogeneous nuclear ribonucleoprotein (hnRNP) B1 protein was found in 100% of squamous cell carcinomas of human lung, we applied the same immunohistochemical method, using anti-hnRNP B1 antibody, to human oral squamous cell carcinoma (OSCC). Seven human tissue sections of OSCC showed strong staining with anti-hnRNP B1 antibody, and hnRNP B1 protein of 37 kDa was identified in protein fractions isolated from six of the cancerous tissue sections, while it was not found in adjacent noncancerous tissue. Moreover, three non-homogeneous (nodular) leukoplakia sections showed significant anti-hnRNP B1 staining. The results suggest that this antibody detects precancerous lesions as well as advanced lesions (stages I to IV) of OSCC. We also present positive results of cytodiagnosis for two smear specimens. All of the above results indicate that hnRNP B1 is a new and useful marker for early detection of OSCC. PMID- 10665655 TI - G1-checkpoint function including a cyclin-dependent kinase 2 regulatory pathway as potential determinant of 7-hydroxystaurosporine (UCN-01)-induced apoptosis and G1-phase accumulation. AB - 7-Hydroxystaurosporine (UCN-01), which was originally identified as a protein kinase C selective inhibitor, is currently in clinical trials as an anti-cancer drug. We previously showed that UCN-01 induced preferential G1-phase accumulation in tumor cells and this effect was associated with the retinoblastoma (Rb) protein and its regulatory factors, such as cyclin-dependent kinase 2 (CDK2) and CDK inhibitors p21Cip1/WAF1 and p27Kip1. We demonstrate here that G1-phase accumulation was induced by UCN-01 in Rb-proficient cell lines (WiDr and HCT116 human colon carcinomas and WI-38 human lung fibroblast), and it was accompanied by dephosphorylation of Rb. In addition, UCN-01-induced G1-phase accumulation was also demonstrated in a Rb-defective cell line (Saos-2 human osteosarcoma), but not in a simian virus 40 (SV40)-transformed cell line (WI-38 VA13). Apoptosis was induced by UCN-01 in the two Rb-deficient cell lines, but not in the other Rb proficient cell lines. These observations suggest that G1-checkpoint function might be important for cell survival during UCN-01 treatment. In addition, there may be a UCN-01-responsive factor in the G1-checkpoint machinery other than Rb which is targeted by SV40. Further studies revealed a correlation between UCN-01 induced G1-phase accumulation and reduction of cellular CDK2 kinase activity. This reduction was strictly dependent on down-regulation of the Thr160 phosphorylated form of CDK2 protein, and coincided in part with up-regulation of p27Kip1, but it was independent of the level of the p21Cip1/WAF1 protein. These results suggest that G1-checkpoint function, including a CDK2-regulatory pathway, may be a significant determinant of the sensitivity of tumor cells to UCN-01. PMID- 10665656 TI - Cisplatin-resistant HeLa cells are resistant to apoptosis via p53-dependent and independent pathways. AB - Since HeLa cells possess very little functional p53 activity, they could be originally resistant to genotoxic stress-induced apoptosis. Therefore, it is likely that the drug-resistant cells derived from HeLa cells are more resistant to apoptosis. The aim of this study was to determine whether cisplatin-resistant cells derived from HeLa cells have an apoptosis-resistant phenotype. A cisplatin resistant cell subline, HeLa/CDDP cells, showed a 19-fold resistance to cisplatin compared with the parent cells. The subline showed a collateral sensitivity to paclitaxel. An equitoxic dose (IC50) of cisplatin produced DNA fragmentation in HeLa cells but not in HeLa/CDDP cells. Transfection of wild-type p53 gene enhanced the cytotoxicity of cisplatin and cisplatin-induced apoptosis in HeLa cells but not in HeLa/CDDP cells, although it caused p53 overexpression in both cell lines. The expression of caspase 1 (interleukin-1beta-converting enzyme, ICE) mRNA and the overexpression of bax protein were observed only in HeLa cells. Paclitaxel-induced DNA fragmentation appeared less in HeLa/CDDP cells than in HeLa cells. p53 gene transfection did not affect the extent of DNA fragmentation in either cell line, suggesting that paclitaxel may induce p53-independent apoptosis. These findings suggest that HeLa/CDDP cells may have an acquired phenotype that is resistant to p53-dependent and -independent apoptosis. PMID- 10665657 TI - Interaction of docetaxel ("Taxotere") with human P-glycoprotein. AB - The interaction of docetaxel ("Taxotere") with P-glycoprotein (P-gp) was examined using porcine kidney epithelial LLC-PK1 and LLC-GA5-COL150 cells, overexpressing human P-gp selectively on the apical plasma membrane by transfection of human MDR1 cDNA into the LLC-PK1 cells. The basal-to-apical transport of [14C]docetaxel in LLC-GA5-COL150 cells significantly exceeded that in LLC-PK1 cells, but the apical-to-basal transport was decreased in LLC-GA5-COL150 cells. The intracellular accumulation after its basal or apical application to LLC-GA5 COL150 cells was 4- to 20-fold lower than that of LLC-PK1 cells. Multidrug resistance (MDR) modulators, i.e., cyclosporin A and SDZ PSC 833, inhibited the basal-to-apical transport and increased the apical-to-basal transport of [14C]docetaxel in LLC-GA5-COL150 cells, but verapamil affected only apical-to basal transport. The intracellular accumulation after basal or apical application to LLC-GA5-COL150 cells was also increased by these three MDR modulators. These observations demonstrated that docetaxel is a substrate for human P-gp, suggesting that docetaxel-drug interactions occur via P-gp. The inhibition of [14C]docetaxel transport by the MDR modulators, as well as daunorubicin and vinblastine, was also found in LLC-PK1 cells, which endogenously express P-gp at lower levels, and concentrations showing similar levels of inhibition were lower than those in the case of LLC-GA5-COL150 cells. These observations indicate that it is necessary to consider the pharmacokinetic and pharmacodynamic interactions of docetaxel via P-gp. PMID- 10665658 TI - Evaluation of antivascular and antimitotic effects of tubulin binding agents in solid tumor therapy. AB - Tubulin binding agents (TBAs) reduce tumor perfusion and inhibit mitosis of tumor cells in solid tumors, but it is not clear which effects contribute to the suppression of solid tumor growth. We evaluated the antivascular and antimitotic effects of several TBAs, combretastatin A-4 (CS A-4) phosphate, AC-7700, a novel CS A-4 derivative, colchicine, E7010, and vinblastine, on subcutaneous (s.c.) murine colon26 adenocarcinoma (c26). Tolerable doses of vinblastine and E7010) strongly inhibited tumor growth and induced mitotic arrest of tumor cells without affecting tumor perfusion. Colchicine had no effect on tumor growth and perfusion. When the injected dose was increased to the lethal range, however, these drugs markedly reduced tumor perfusion and caused necrosis of tumor tissue. Within the tolerable dose range, AC-7700 both strongly suppressed tumor growth and reduced tumor perfusion, and CS A-4 phosphate also exhibited a moderate antivascular effect. To evaluate the contribution of antivascular activity of TBAs to tumor growth suppression, excluding their direct cytotoxic effect on tumor cells, we established c26/acr, which is resistant to TBAs in vitro. Although E7010 showed a reduced suppressive effect on s.c. c26/acr tumor growth as compared with its effect on wild-type c26, AC-7700 remained potent against both cell lines. These results indicate that TBAs exert antivascular and antimitotic effects on solid tumors with marked differently effective dose ranges from agent to agent, and that the antivascular effect of TBAs inhibits solid tumor growth independently of the direct cytotoxic effect on tumor cells. PMID- 10665659 TI - Clinical teratology: identifying teratogenic risks in humans. PMID- 10665660 TI - Genetic landmarks through philately--Henry Louis 'Lou' Gehrig and amyotrophic lateral sclerosis. PMID- 10665661 TI - Human renin gene BglI dimorphism associated with hypertension in two independent populations. AB - The renin (REN) gene is a good candidate that could underlie an individual's genetic susceptibility to human essential hypertension (EHT). We describe here a polymerase chain reaction-based assay for detection of a BglI dimorphic site located in the first intron of the REN gene. In this retrospective, case-control, association study, we investigated BglI allele and genotype distributions in 554 subjects (280 hypertensives and 274 normotensives) from the United Arab Emirates (UAE) - a genetically homogeneous ethnic population with no history of smoking or alcohol consumption - and in 485 hypercholesterolemic, US Caucasian subjects (250 hypertensives and 235 normotensives). A statistically significant association was found between alleles on which the BglI site is present [BglI(+)] and clinical diagnosis of EHT in the UAE sample group (odds ratio = 2.69, p = 0.0006), and a similar trend was observed in the US group (odds ratio = 1.97, p = 0.01). BglI(+) homozygous status was also investigated in the US group and found to be associated with elevated systolic and diastolic blood pressure values (respectively, 144.8+/-26.1 vs. 134.1+/-23.0 mm Hg, p = 0.04; and 91.0+/-12.5 vs. 82.2+/-12.7 mm Hg, p = 0.009). In conclusion, variations of the REN (or of a nearby) gene that may be in linkage disequilibrium with the REN BglI(+) marker could play a role in contributing to an increased individual's genetic susceptibility to EHT in the UAE population and amongst US hypercholesterolemic Caucasians. Such a genetic influence, which seems to show a recessive mode of inheritance, could also be implicated in raising both systolic and diastolic blood pressures. PMID- 10665662 TI - Perceptions of the outcome of orthopedic surgery in patients with chondrodysplasias. AB - As part of a larger survey of patients with chondrodysplasias, 197 patients or their parents were asked whether they had undergone orthopedic surgery related to their chondrodysplasia and, if so, to rate their impression of the outcome. Seventy-four patients (37.6%) had undergone a total of 152 procedures (221 if concurrent bilateral operations are counted separately). The percentage of patients treated surgically ranged from a low of 8.3% for hypochondroplasia to a high of 87.5% for diastrophic dysplasia. Of the patients who had surgery, the mean number of procedures per patient ranged from 1.0 for hypochondroplasia to 2.69 for pseudoachondroplasia. Of 180 individual procedures related to the limbs, the outcome in 88.8% was judged 'a bit better' or higher and in 68.8% 'much better' or higher. The responses ranged from a low of 70.4 and 66.7%, respectively for proximal femoral osteotomies to a high of 100 and 85.9% for hip replacement. The comparable figures for spine related surgery were 81.8 and 48.5% with a low of 58.3 and 50.0% for foramen magnum-cervical surgery and a high of 93.8 and 43.8% for thoracolumbar procedures. The expressed perception of lack of satisfaction varied not only by procedure but by diagnosis. Overall, patients perceived a high level of post-surgical improvement, although a number experienced subsequent deterioration and the need for further intervention. PMID- 10665663 TI - Molecular genetics of Turner syndrome: correlation with clinical phenotype and response to growth hormone therapy. AB - To correlate the origin of the retained X in Turner syndrome with phenotype, pre treatment height and response to recombinant human growth hormone (rhGH) therapy, systematic clinical assessment and molecular studies were carried out in 33 Greek children with Turner syndrome and their parents including 18 children with 45,X and 15 with X-mosaicism. Microsatellite markers on X chromosomes (DXS101 and DXS337) revealed that the intact X was paternal (Xp) in 15/30 and maternal (Xm) in 15/30 children, while 3/33 families were non-informative. No significant relationship was found between parental origin of the retained X and birth weight/length/gestational age, blepharoptosis, pterygium colli, webbed neck, low hairline, abnormal ears, lymphoedema, short 4th metacarpal, shield chest, widely spaced nipples, cubitus valgus, pigmented naevi, streak gonads, and cardiovascular/renal anomalies. With regard to the children's pre-treatment height, there was a significant correlation with maternal height and target height in both Xm and Xp groups. No differences were found between Xm and Xp groups and the improvement of growth velocity (GV) during the first and second year of rhGH administration, while for both groups GV significantly improved with rhGH by the end of the first and the second year. To our knowledge, this is the first attempt to correlate the parental origin of Turner syndrome with the response to rhGH therapy. PMID- 10665664 TI - Relation of mathematical ability to psychosis in Iceland. AB - A study of mathematically gifted Icelanders demonstrates an increased risk of mental illness in their ranks. Psychotic disorders are also frequent among their relatives. Linkage of academic records with previously published data on the family distribution of psychosis reveals a pattern compatible with the hypothesis that high arousal plays a role in reasoning ability. PMID- 10665665 TI - Prevalence of human leukocyte antigen DQA1 and DQB1 alleles in Kuwaiti Arab children with type 1 diabetes mellitus. AB - The prevalence of human leukocyte antigen (HLA) DQB1 and DQA1 alleles has been determined in 78 Kuwaiti Arab children with insulin-dependent diabetes mellitus (IDDM) and in 57 normal healthy controls with similar ethnic background. The typing of HLA-DQ alleles was carried out using an allele-specific DNA-based polymerase chain reaction (PCR) SSP method. DR typing was also performed in 212 control subjects using PCR-SSP (sequence specific primer) method. A significantly higher frequency of DQB1*0201 allele was found in IDDM cases compared to the controls (p<0.001). There was no significant difference in the prevalence of DQB1 alleles *0302, *0501, and *0602 between IDDM cases and the controls. In contrast, DQB1 alleles *0301, *0402, *0502, *0602, and *0603 were represented at a somewhat higher frequency in controls compared to the IDDM cohort. The frequency of DQA1 allele *0301, which encode for an Arg at codon 52, was significantly higher in the IDDM patients compared to the controls (p<0.001). The frequency of DQA1 allele *0302 was also higher in IDDM cases than controls (p = 0.034) but the difference was less pronounced than DQA1*0301. Amongst the Arg52 alleles, no significant difference was detected in the frequency of *0401 between IDDM cases and the controls and the allele *0501 was detected only in controls. For non Arg52 alleles *0103, *0104, and *0201, the differences in the two groups were not significant, with the exception of allele *0104 (p = 0.024). DR3 was the most common type in the Kuwaiti general population (28%) and DRB1*0301 was detected in 41% of the individuals with DR3 specificity. Analysis of HLA-DQBI/DQA1 haplotypes from IDDM cases and controls revealed a significantly high frequency of haplotype DQA1*0301/DQB1*0201 between Kuwaiti IDDM cases (49/78, 63%) and the controls (8/57, 14%). PMID- 10665666 TI - Proximal myotonic myopathy: clinical and molecular investigation of a Norwegian family with PROMM. AB - Proximal myotonic myopathy (PROMM) was first described in 1994 as a multisystem disorder with similarity to myotonic dystrophy (DM), but without the abnormal (CTG)n expansion in the DM protein kinase (DMPK) gene. The inheritance is autosomal dominant and the clinical features include myotonia, proximal muscle weakness and cataract. Linkage analysis in nine German PROMM families has indicated the possibility of linkage to DM2 locus on chromosome 3. We report a Norwegian PROMM family in which the proband was clinically diagnosed as DM but without the (CTG)n expansion. Using an intragenic marker we showed that the DMPK gene did not segregate with the disease in this family. All family members are heterozygous for the R894X mutation in CLCN1 gene. Linkage analysis could not be performed, but haplotyping probably excludes the DM2 locus as the disease locus in this family. The present family emphasises that myalgia is a prominent symptom in PROMM and the clinical differences may be explained by genetic heterogeneity. This family will be reinvestigated along with the identification of candidate genes or regions in larger PROMM families. PMID- 10665667 TI - Postaxial polydactyly and Dandy-Walker malformation. Further nosological comments. PMID- 10665668 TI - Search for uniparental disomy 14 in balanced Robertsonian translocation carriers. PMID- 10665669 TI - Prenatal diagnosis of peroxisome biogenesis disorders by means of immunofluorescence staining of cultured chorionic villous cells. PMID- 10665670 TI - A de novo heterozygous deletion of 42 base-pairs in the noggin gene of a fibrodysplasia ossificans progressiva patient. PMID- 10665671 TI - Sensory feedback in artificial control of human mobility. AB - Artificial motor control systems may reduce the handicap of motor impaired individuals. Sensors are essential components in feedback control of these systems and in the information exchange with the user. The objective of this paper is to give an overview of the applications of sensors in the artificial control of human mobility. These applications may either require an accurate estimate of the measured physical quantities or can be based on learning the relation between sensory information and control actions by example. Actual use of sensors in artificial motor control systems requires that the user experienced complexity of the system is not increased, while improving the repeatable and flexible functioning of the system. Therefore, the sensors need to be integrated with the mechanical part of the artificial support system or implanted, the information exchange between sensor and controller should be wireless and automatic in-use calibration is a desired feature. PMID- 10665672 TI - Interfacing the body's own sensing receptors into neural prosthesis devices. AB - Functional Electric Stimulation (FES) is today available as a tool in muscle activation used in picking up objects, in standing and walking, in controlling bladder emptying, and for breathing. Despite substantial progress over nearly three decades of development, many challenges remain to provide a more efficient functionality of FES systems. The most important of these is an improved control of the activated muscles. Instead of artificial sensors for feedback, new developments in electrodes to do long-term and reliable recordings from peripheral nerves emphasize the use of the body's own sensors. These are already installed and optimised through millions of years of natural evolution. This paper presents recent results on a system using electrical stimulation of motor nerves to produce movement and using the natural sensors as feedback signals to control the stimulation that can replicate some of the functions of the spinal cord and its communication with the brain. We have used the nerve signal recorded from cutaneous nerves in two different human applications: (1) to replace the external heel switch of a system for correction of spastic drop foot by peroneal stimulation, and (2) to provide an FES system for restoration of hand grasp with sensory feedback from the fingertip. For the bladder function, the sacral root stimulator is a useful control tool in emptying the bladder. To decide when to stimulate, we are at present carrying out experiments on pigs and cats using cuff electrodes on the pelvic nerve and sacral roots to record the neural information from bladder afferents. This information can potentially be used to inhibit unwanted bladder contractions and to trigger the FES system and thereby bladder emptying. Future research will show whether cuffs and other types of electrodes can be used to reliably extract signals from the large number of other receptors in the body to improve and expand on the use of natural sensors in clinical FES systems. PMID- 10665673 TI - Strategies for providing upper extremity amputees with tactile and hand position feedback--moving closer to the bionic arm. AB - A continuing challenge for prostheses developers is to replace the sensory function of the hand. This includes tactile sensitivity such as finger contact, grip force, object slippage, surface texture and temperature, as well as proprioceptive sense. One approach is sensory substitution whereby an intact sensory system such as vision, hearing or cutaneous sensation elsewhere on the body is used as an input channel for information related to the prosthesis. A second technique involves using electrical stimulation to deliver sensor derived information directly to the peripheral afferent nerves within the residual limb. Stimulation of the relevant afferent nerves can ultimately come closest to restoring the original sensory perceptions of the hand, and to this end, researchers have already demonstrated some degree of functionality of the transected sensory nerves in studies with amputee subjects. This paper provides an overview of different types of nerve interface components and the advantages and disadvantages of employing each of them in sensory feedback systems. Issues of sensory perception, neurophysiology and anatomy relevant to hand sensation and function are discussed with respect to the selection of the different types of nerve interfaces. The goal of this paper is to outline what can be accomplished for implementing sensation into artificial arms in the near term by applying what is present or presently attainable technology. PMID- 10665674 TI - Robots with a gentle touch: advances in assistive robotics and prosthetics. AB - As healthcare costs rise and an aging population makes an increased demand on services, so new techniques must be introduced to promote an individuals independence and provide these services. Robots can now be designed so they can alter their dynamic properties changing from stiff to flaccid, or from giving no resistance to movement, to damping any large and sudden movements. This has some strong implications in health care in particular for rehabilitation where a robot must work in conjunction with an individual, and might guiding or assist a persons arm movements, or might be commanded to perform some set of autonomous actions. This paper presents the state-of-the-art of rehabilitation robots with examples from prosthetics, aids for daily living and physiotherapy. In all these situations there is the potential for the interaction to be non-passive with a resulting potential for the human/machine/environment combination to become unstable. To understand this instability we must develop better models of the human motor system and fit these models with realistic parameters. This paper concludes with a discussion of this problem and overviews some human models that can be used to facilitate the design of the human/machine interfaces. PMID- 10665675 TI - Overview of clinical trials with MIT-MANUS: a robot-aided neuro-rehabilitation facility. AB - We are applying robotics and information technology to assist, enhance, and quantify neuro-rehabilitation. Our goal is a new class of interactive, user affectionate clinical devices designed not only for evaluating patients, but also for delivering meaningful therapy via engaging "video games". Notably, the novel robot MIT-MANUS has been designed and programmed for clinical neurological applications, and has undergone extensive clinical trials for more than four years at Burke Rehabilitation Hospital - White Plains, NY. This paper will review results of the first clinical trial of 20 patients, which showed that: - Stroke patients treated daily with additional robot-aided therapy during acute rehabilitation had improved outcome in motor activity at hospital discharge, when compared to a control group that received only standard acute rehabilitation treatment. - This improved outcome was sustained after three years. - The neuro recovery process continued far beyond the commonly accepted 3 months post-stroke interval. PMID- 10665676 TI - Human--machine load sharing in rehabilitation robotics. AB - A force-assist mechanism has been developed to mount on the Chameleon - a wheelchair mounted rehabilitation robot. The device will amplify the forces applied by the user, making it possible to lift a large weight with a small force. This paper describes the test-bed development; instrumentation of the Chameleon with power assistance; and preliminary results on system efficacy. PMID- 10665677 TI - Mechatronic assessment of arm impairment after chronic brain injury. AB - Significant potential exists for mechatronic devices to improve assessment and treatment of individuals with a movement disability following stroke, traumatic brain injury, or cerebral palsy. We report the use of a mechatronic device for evaluation of the arm after chronic brain injury. We performed a series of experiments with the device in order to identify the relative contribution of three different motor impairments to decreased active range of motion of reaching in five brain-injured subjects. Our findings were that passive tissue restraint and agonist weakness, rather than antagonist restraint, were the most common contributors to decreased active range of motion. These results demonstrate the feasibility of objective assessment of functional movement using a mechatronic device, and could provide the basis for improved, individualized treatment planning and monitoring following brain injury. PMID- 10665678 TI - Postural activity of constrained subject in response to disturbance in sagittal plane. AB - The study examines postural activity of a constrained subject in response to various disturbances in the sagittal plane. Three different initial standing postures were examined. Each response to a perturbation was divided into three consecutive phases according to the intensity of the muscular activity. The relation of the ankle joint torque versus the ankle joint angle was studied. A linear relationship, resulting in constant ankle stiffness, was observed in each phase of the response. Only negligible differences in the stiffness values were observed among different phases. The results indicate an ankle stiffness value of 10 N m/o for the majority of initial stance postures and perturbation intensities. PMID- 10665679 TI - Feedback control of unsupported standing. AB - This paper presents the results of continuing work on feedback control of unsupported standing in paraplegia. Our experimental setup considers a situation in which all joints above the ankle are braced, and stabilising torque at the ankle is generated by stimulation of the plantarflexors. A previous study showed that short periods of unsupported standing with paraplegic subjects could be achieved. In order to improve consistency and reliability of unsupported standing we are currently investigating several modifications to the control strategy. The paper reports progress towards this goal. PMID- 10665680 TI - Towards applicable ballistic walking. AB - Ballistic walking has been a topic of research in biomechanics and robotics several times. The main advantages of this approach are: self-organizing properties, minimal energetic expenditure, and natural movement. This would make ballistic walking a perfect physical principle for application in rehabilitation, and robotics. However a typical shortcoming of this approach is that it does not explicitly involve actuation (e.g., muscles or motors). In this work ballistic walking is extended with variable intrinsic system parameters, which can be used as an energy input and for disturbance control. With a limit cycle approach walking cycles can be synthesized and cycle controllers can be designed. Experiments with a bipedal robot and simulations illustrate the route towards more robust, and applicable ballistic walking. PMID- 10665681 TI - Estimating orientation with gyroscopes and accelerometers. PMID- 10665682 TI - Design of an intramedullary leg lengthening device with a shape memory actuator. AB - The procedure and the external fixator for lengthening long bones was developed by G.A. Ilizarov in the late 1960's. This technique has, despite its proven abilities for leg lengthening and correction of angular deformities, some considerable disadvantages for the patients. Discomfort, infections and restricted weight bearing are some reasons for the development of a completely intramedullary device for leg lengthening. The device developed at the Laboratory of Biomechanical Engineering, University of Twente, is a telescopic intramedullary nail with a maximum diameter of 13 mm, which can be lengthened with 0.5 mm steps induced by a shape memory alloy actuator. The electrical energy for the actuator is supplied from outside the body by inductive coupling of two solenoid coils. Internally, the electrical energy is transformed to thermal energy by Thermofoils and Peltier-elements. PMID- 10665684 TI - Gene therapy trials show clinical efficacy. PMID- 10665683 TI - Good Fridays. PMID- 10665686 TI - An ancient nation braces to fight AIDS. PMID- 10665685 TI - Some men who take Viagra die--why? PMID- 10665687 TI - From the Food and Drug Administration. PMID- 10665688 TI - From the Centers for Disease Control and Prevention. Staphylococcus aureus with reduced susceptibility to vancomycin--Illinois, 1999. PMID- 10665689 TI - From the Centers for Disease Control and Prevention. Alcohol involvement in fatal motor-vehicle crashes--United States, 1997-1998. PMID- 10665690 TI - From the Centers for Disease Control and Prevention. Laboratory capacity to detect antimicrobial resistance, 1998. PMID- 10665691 TI - Primary angioplasty vs thrombolysis in elderly patients. PMID- 10665692 TI - Quality of health care and the HMO marketplace. PMID- 10665693 TI - Quality of health care and the HMO marketplace. PMID- 10665694 TI - Quality of health care and the HMO marketplace. PMID- 10665695 TI - Quality of health care and the HMO marketplace. PMID- 10665696 TI - Quality of health care and the HMO marketplace. PMID- 10665697 TI - Glycosylated hemoglobin as a diagnostic test for type 2 diabetes mellitus. PMID- 10665698 TI - Glycosylated hemoglobin as a diagnostic test for type 2 diabetes mellitus. PMID- 10665699 TI - Glycosylated hemoglobin as a diagnostic test for type 2 diabetes mellitus. PMID- 10665700 TI - Long-term outcomes of persons with Lyme disease. AB - CONTEXT: Few data exist about the long-term outcomes of patients with Lyme disease. OBJECTIVE: To assess the long-term outcomes of patients with Lyme disease. DESIGN: Two-part project including a community-based longitudinal cohort study and a matched cohort study. SETTING AND PARTICIPANTS: Six hundred seventy eight patients identified from a random sample of all reports of Connecticut residents with suspected Lyme disease submitted to the Connecticut Department of Public Health from 1984-1991 were evaluated in the longitudinal study; for a random subsample of 212 patients from the larger study, 212 age-matched controls without Lyme disease also were enrolled. MAIN OUTCOME MEASURES: Self-reports or parents' reports of symptoms and ability to perform certain daily activities since diagnosis of Lyme disease; scores on the 36-Item Short-Form Health Survey and the Center for Epidemiologic Studies-Depression scale, for adults, by case definition status and between patients and controls. RESULTS: Of the 678 patients, 51.6% were female, 34.4% were children, and 64.3% met the national surveillance case definition for Lyme disease. Most patients (85.6%) were treated with antimicrobial agents. Interviews were conducted a median of 51 months after diagnosis (range, 15-135 months). An increased frequency of symptoms (eg, pain, fatigue) or of difficulty with daily activities (eg, performing housework, exercising) was reported by 69% of the patients, although few (19%) of these problems were attributed to Lyme disease. Whenever there was a statistically significant difference in the frequencies of either increased symptoms or increased difficulties with typical activities between those who did or did not meet the surveillance case definition, in all instances the greater frequency of problems was in the group that did not meet the case definition. The frequencies of reports of both increased symptoms and increased difficulties with typical activities among patients who had been diagnosed as having Lyme disease were similar to those among age-matched controls without Lyme disease. CONCLUSIONS: In this cohort, although many patients reported increases in symptoms and/or increased difficulties with typical daily activities between 1 and 11 years after diagnosis of Lyme disease, the frequencies of these reports were similar to the frequencies of such reports among age-matched controls without Lyme disease. PMID- 10665701 TI - Life expectancy gains from cancer prevention strategies for women with breast cancer and BRCA1 or BRCA2 mutations. AB - CONTEXT: Women with BRCA1- or BRCA2-associated breast cancer are at increased risk for contralateral breast cancer and ovarian cancer and therefore may consider secondary cancer prevention strategies, such as prophylactic surgery and tamoxifen therapy. It is not proven to what extent these strategies reduce risk of second cancers in such patients. OBJECTIVE: To examine the effect of tamoxifen therapy, bilateral prophylactic oophorectomy (PO), prophylactic contralateral mastectomy (PCM), and combinations of these strategies on life expectancy for women with unilateral breast cancer and a BRCA1 or BRCA2 gene mutation. DESIGN AND SETTING: Decision analysis using a Markov model. Probabilities for developing contralateral breast cancer and ovarian cancer, dying from these cancers, dying from primary breast cancer, and the reduction in cancer incidence and mortality due to prophylactic surgeries and/or tamoxifen were estimated from published studies. PARTICIPANTS: Hypothetical breast cancer patients with BRCA1 or BRCA2 mutations facing decisions about secondary cancer prevention strategies. INTERVENTIONS: Seven strategies, including 5 years of tamoxifen use, PO, PCM, and combinations of these strategies, compared with careful surveillance. MAIN OUTCOME MEASURES: Total and incremental life expectancy (LE) with each intervention strategy. RESULTS: Depending on the assumed penetrance of the BRCA mutation, compared with surveillance alone, 30-year-old early-stage breast cancer patients with BRCA mutations gain in LE 0.4 to 1.3 years from tamoxifen therapy, 0.2 to 1.8 years from PO, and 0.6 to 2.1 years from PCM. The magnitude of these gains is least for women with low-penetrance mutations (assumed contralateral breast cancer risk of 24% and ovarian cancer risk of 6%) and greatest for those with high-penetrance mutations (assumed contralateral breast cancer risk of 65% and ovarian cancer risk of 40%.) Older age and poorer prognosis from primary breast cancer further attenuate these gains. CONCLUSIONS: Interventions to prevent second cancers, particularly PCM, may offer substantial LE gain for young women with BRCA-associated early-stage breast cancer. Estimates of LE gain may help women and their physicians consider the uncertainties, risks, and advantages of these interventions and lead to more informed choices about cancer prevention strategies. PMID- 10665702 TI - Adult functional outcome of those born small for gestational age: twenty-six-year follow-up of the 1970 British Birth Cohort. AB - CONTEXT: Although studies have documented cognitive impairment in children who were born small for gestational age (SGA), other studies have not demonstrated differences in IQ or other cognitive scores. The need exists for long-term studies of such children to assess functional outcomes not measurable with standardized testing. OBJECTIVE: To determine the long-term functional outcome of SGA infants. DESIGN: Prospective cohort study. SETTING AND PARTICIPANTS: A total of 14,189 full-term infants born in the United Kingdom on April 5 through 11, 1970, were studied as part of the 1970 British Birth Cohort; 1064 were SGA (birth weight less than the fifth percentile for age at term). Follow-up at 5, 10, 16, and 26 years was 93%, 80%, 72%, and 53%, respectively. MAIN OUTCOME MEASURES: School performance and achievement, assessed at 5, 10, and 16 years; and years of education, occupational status, income, marital status, life satisfaction, disability, and height, assessed at 26 years, comparing persons born SGA with those who were not. RESULTS: At 5, 10, and 16 years of age, those born SGA demonstrated small but significant deficits in academic achievement. In addition, teachers were less likely to rate those born SGA in the top 15th percentile of the class at 16 years (13% vs 20%; P<.01) and more likely to recommend special education (4.9% vs 2.3%; P<.01) compared with those born at normal birth weight (NBW). At age 26 years, adults who were SGA did not demonstrate any differences in years of education, employment, hours of work per week, marital status, or satisfaction with life. However, adults who were SGA were less likely to have professional or managerial jobs (8.7% vs 16.4%; P<.01) and reported significantly lower levels of weekly income (mean [SD], 185 [91] vs 206 [102] pound sterling; P<.01) than adults who were NBW. Adults who were SGA also reported significant height deficits compared with those who were NBW (mean [SD] z score, -0.55 [0.98] vs 0.08 [1.02]; P<.001). Similar results were also obtained after adjusting for social class, sex, region of birth, and the presence of fetal or neonatal distress. CONCLUSIONS: In this cohort, adults who were born SGA had significant differences in academic achievement and professional attainment compared with adults who were NBW. However, there were no long-term social or emotional consequences of being SGA: these adults were as likely to be employed, married, and satisfied with life. PMID- 10665703 TI - Arterial hypertension and renal allograft survival. AB - CONTEXT: Several observational studies have investigated the significance of hypertension in renal allograft failure; however, these studies have been complicated by the lack of adjustment for baseline renal function, leaving the role of elevated blood pressure in allograft failure unclear. OBJECTIVE: To examine the relationship between blood pressure adjusted for renal function and survival after cadaveric allograft transplantation. DESIGN: Nonconcurrent historical cohort study conducted from 1985 through 1997. SETTING: University teaching hospital. PARTICIPANTS: A total of 277 patients aged 18 years or older who underwent cadaveric renal transplantation without another simultaneous organ transplantation and whose allograft was functioning for a minimum of 1 year. Follow-up continued through 1997 (mean follow-up, 5.7 years). MAIN OUTCOME MEASURE: Time to allograft failure (defined as death, return to dialysis, or retransplantation) by systolic, diastolic, and mean arterial blood pressure measurements at 1 year after transplantation. RESULTS: Multivariate Cox proportional hazards modeling demonstrated that nonwhite ethnicity, history of acute rejection, and nondiabetic kidney disease were significant predictors of failure (P = .01 for all). In addition, the calculated creatinine clearance at 1 year had an adjusted rate ratio (RR) for allograft failure per 10 mL/min (0.17 mL/s) of 0.74 (95% confidence interval [CI], 0.62-0.88). The RR per 10-mm Hg increase in blood pressure measured at 1 year after transplantation, after adjustment for creatinine clearance, was 1.15 (95% CI, 1.02-1.30) for systolic pressure, 1.27 (95% CI, 1.01-1.60) for diastolic pressure, and 1.30 (95% CI, 1.05 1.61) for mean arterial pressure. Supplemental analyses that did not include death as a failure event or reduce the minimum allograft survival time for study subjects to 6 months yielded results consistent with the primary analysis. There was no evidence of modification of the blood pressure-allograft failure relationship by ethnicity or diabetes mellitus. CONCLUSIONS: Systolic, diastolic, and mean arterial blood pressures at 1 year posttransplantation strongly predict allograft survival adjusted for baseline renal function. More aggressive control of blood pressure may prolong cadaveric allograft survival. PMID- 10665704 TI - The role of clinical suspicion in evaluating a new diagnostic test for active tuberculosis: results of a multicenter prospective trial. AB - CONTEXT: In laboratory trials, nucleic acid amplification tests for the diagnosis of tuberculosis (TB) are more accurate than acid-fast bacilli (AFB) smear microscopy and are faster than culture. The impact of these tests on clinical diagnosis is not known. OBJECTIVE: To assess the performance of a nucleic acid amplification test, the enhanced Mycobacterium tuberculosis Direct (E-MTD) test, against a uniform clinical standard stratified by level of clinical suspicion. DESIGN: Prospective multicenter trial conducted between February and December 1996, documenting the clinical suspicion of TB at enrollment and using final comprehensive diagnosis as the criterion standard. SETTING: Six urban medical centers and 1 public health TB clinic. PATIENTS: A total of 338 patients with symptoms and signs consistent with active pulmonary TB and complete clinical diagnosis were stratified by the clinical investigators to be at low (< or =25%), intermediate (26%-75%), or high (>75%) relative risk of having TB. MAIN OUTCOME MEASURES: Sensitivity, specificity, and positive and negative predictive values of the E-MTD test in clinical suspicion of groups with low (n = 224); intermediate (n = 68); and high (n = 46) clinical suspicion of TB. RESULTS: Based on comprehensive clinical diagnosis, sensitivity of the E-MTD test was 83%, 75%, and 87% for low, intermediate, and high clinical suspicion of TB, respectively, and corresponding specificity was 97%, 100%, and 100% (P = .25). Positive predictive value of the E-MTD test was 59% (low), 100% (intermediate), and 100% (high) compared with 36% (low), 30% (intermediate), and 94% (high) for AFB smear. Corresponding negative predictive values were 99%, 91%, and 55% [corrected] (E MTD test) vs 96%, 71%, and 37% (AFB smear). CONCLUSIONS: For complex diagnostic problems like TB, clinical risk assessments can provide important information regarding predictive values more likely to be experienced in clinical practice. For this series, a clinical suspicion of TB was helpful in targeting areas of the clinical spectrum in which nucleic acid amplification tests can make an important contribution. PMID- 10665705 TI - Sex differences in evaluation and outcome of unstable angina. AB - CONTEXT: The existence of sex bias in the delivery of cardiac care is controversial, and little is known about the association between sex and delivery of care and outcomes at an early point in the diagnostic sequence, such as when patients present for the evaluation of chest pain. OBJECTIVE: To test the hypothesis that female sex is negatively associated with care delivered to and outcomes of persons diagnosed as having unstable angina. DESIGN: Inception population-based cohort study with an average of 6 years of follow-up. SETTING: Emergency departments (EDs) in Olmsted County, Minnesota. PATIENTS: A total of 2271 Olmsted County residents (1306 men and 965 women) who presented to the ED for the first time with symptoms meeting criteria for unstable angina between 1985 and 1992. MAIN OUTCOME MEASURES: Use of cardiac procedures within 90 days of ED visit, overall mortality, and cardiac events (cardiac death, nonfatal myocardial infarction, nonfatal cardiac arrest, and congestive heart failure), compared by sex and Agency for Health Care Policy and Research cardiovascular risk category (low, intermediate, or high). RESULTS: Women were older (P<.001), more likely to have a history of hypertension (P = .001), and less likely to present with typical angina (P = .004) than men. Men were more likely than women to undergo noninvasive cardiac tests (relative risk [RR], 1.27; 95% confidence interval [CI], 1.14-1.40) as well as invasive cardiac procedures (RR, 1.72; 95% CI, 1.51-1.97). After adjustment, male sex was associated with a 24% increase in the use of cardiac procedures. Survival of both men and women in the high and intermediate risk categories was significantly lower than expected per the general population (P<.001). Women had a worse outcome than men, but after multivariate adjustment, male sex was associated with a trend toward an increase in the risk of death (RR, 1.23; 95% CI, 0.99-1.54) and significantly associated with increased risk of cardiac events (RR, 1.21; 95% CI, 1.03-1.42). CONCLUSIONS: Our population-based data indicate that after an ED visit for symptoms of unstable angina, the use of cardiac procedures was lower in women, but after taking into account baseline characteristics, men experienced worse outcomes. PMID- 10665706 TI - Cytomegalovirus retinitis in the era of highly active antiretroviral therapy. AB - A number of striking changes have occurred recently in the presentation and course of cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome (AIDS) who are receiving highly active antiretroviral therapy (HAART). Before the use of HAART, CMV retinitis was the most common intraocular infection in patients with AIDS, occurring in up to 40% of patients, typically when CD4+ cell counts have decreased to less than 0.10 x 10(9)/L. By studying CMV retinitis, clinicians can investigate whether the rejuvenated immune system that results from HAART can effectively control opportunistic infections in patients with AIDS. In some patients, retinitis has not progressed when specific anti-CMV therapy was discontinued, but a number of patients have developed substantial intraocular inflammation, which has resulted in decreased visual acuity. Anterior uveitis, cataract, vitritis, cystoid macular edema, epiretinal membrane, and disc edema may occur in patients with CMV retinitis who have experienced HAART-associated elevation in CD4+ cell counts. Since immune recovery uveitis does not occur in eyes without CMV retinitis, the ocular inflammation appears to be related to the CMV infection. Anti-CMV maintenance therapy likely can be safely discontinued in some patients with CMV retinitis if CD4+ cell counts are stable or increasing and have been higher than 0.10 x 10(9)/L for at least 3 months. Immune recovery in patients receiving HAART has been effective in controlling opportunistic infections, but it may also result in intraocular inflammation, which can have adverse effects on the eye. PMID- 10665707 TI - Long-term outcomes and management of patients with Lyme disease. PMID- 10665708 TI - Sex bias in cardiovascular care: should women be treated more like men? PMID- 10665709 TI - The feminization of medicine. PMID- 10665710 TI - Critical women's health issues in the 21st century. PMID- 10665712 TI - Why aren't there more women surgeons?. Interview by Valerie A. Jones. PMID- 10665711 TI - Saints and sinners: women and the practice of medicine throughout the ages. PMID- 10665713 TI - Encouraging the advancement of women. PMID- 10665714 TI - JAMA Patient Page: Lyme disease. PMID- 10665715 TI - A manuscript worth a villa: Vittorio Putti's acquisition of the Guy de Chauliac manuscript. PMID- 10665716 TI - Clinical course in synovial sarcoma: a Scandinavian sarcoma group study of 104 patients. AB - We analyzed treatment and outcome in 104 Scandinavian patients with synovial sarcoma in the extremities or trunk wall, diagnosed between 1986 and 1994. Only surgically treated patients without metastases at diagnosis were included. Median follow-up of survivors was 6 (3-11) years. 34 patients developed metastases. The overall 5- and 7-year survival rates were 0.76 (95% CI 0.66-0.83) and 0.69 (0.58 0.78), respectively. Large tumor size and amputation were significantly associated with impaired metastasis-free survival. Patients with local recurrence had a higher risk of metastases following the local event. Local excision with inadequate margin was associated with a higher risk of local recurrence. PMID- 10665717 TI - Synovial sarcoma--identification of favorable and unfavorable histologic types: a Scandinavian sarcoma group study of 104 cases. AB - Synovial sarcoma has traditionally been regarded as a high-grade sarcoma and treated as such. Recently, specific types of poorly differentiated synovial sarcoma have been defined and shown to affect prognosis adversely. We studied 104 primary synovial sarcomas of the extremities and trunk wall without metastasis at diagnosis that were retrieved from the Scandinavian Sarcoma Group Registry (SSG) and the Swedish Cancer Registry from 1986 to 1994. Follow-up was available in all patients, median 6 (3-11) years for the survivors. There were local recurrences in 15% of patients and metastases in 33%. Histologically, the tumors were divided into favorable and unfavorable types. The favorable type had no significant cytologic atypia, and in most instances, no necrosis and a mitotic count of < 10/10 hpf. The unfavorable type included so-called poorly differentiated synovial sarcomas as well as recognizable biphasic and monophasic synovial sarcomas with prominent nuclear atypia, extreme cellularity and nuclear crowding. Designation of a tumor as having favorable vs. unfavorable histology conveyed more prognostic information than any single histologic factor. Kaplan-Meier estimates of metastasis-free survival at 5 years were 83% for patients with histologically favorable tumors and 31% for patients with histologically unfavorable tumors (95% confidence intervals 72-92% and 13-51%, respectively). These findings may influence future treatment protocols for synovial sarcoma. PMID- 10665718 TI - Incomplete incorporation of morselized and impacted autologous bone graft: a histological study in 4 intracorporally grafted lumbar fractures. AB - Morselized and impacted bone allografts are used successfully in hip and knee revisions, but experiments using bone chambers indicate that impaction actually can delay ingrowth of new bone into a graft. To understand the remodeling and incorporation process of morselized and impacted grafts, we studied the incorporation of morselized impacted autografts in lumbar fractures histologically. 4 patients were operated on for Th XII-LI fractures. The fractures were stabilized by VSP plates and transpedicular screws in the vertebrae above and below the fractured one. Autologous bone graft was packed into the fractured vertebral body through one of the pedicles. After 18-20 months, the plates were removed and biopsies were obtained from various locations in the fractured vertebra. All fractures were at this time clinically and radiographically healed. Histologically, in all cases, large areas of the autograft in the vertebral body were unvascularized and partially or entirely necrotic. As with morselized bone in hip revisions, evaluation of graft incorporation requires histological examination. Full osseous incorporation of a graft is not always necessary for a good clinical result. PMID- 10665719 TI - OP-1 for cervical spine fusion: bridging bone in only 1 of 4 rheumatoid patients but prednisolone did not inhibit bone induction in rats. AB - We used OP-1 (also called BMP-7) on a collagen type-1 carrier in atlanto-axial posterior fusions to promote bony healing after wire fixation. 4 patients who had instability between the atlas and axis due to rheumatoid disease received the implants. The patients were examined with conventional radiography postoperatively at 2, 6 and 10 months. In 3 patients, no new bone formation was detectable. In 1 patient, new bone bridged the fusion site at 6 months. 3 patients were on chronic steroid treatment, including the patient in whom bone formation was detected. To determine whether steroid treatment could be responsible for the low rate of bone induction, 24 rats each received OP-1 implants in an abdominal muscle pouch. They were divided into 3 groups receiving saline, 0.1 or 1.0 mg/kg BW of prednisolone daily until they were killed 3 weeks postoperatively. Specimens were decalcified for histology and the amount of calcium in the decalcifying solution was measured. All groups showed ossicles induced by OP-1, and no effect of prednisolone was detected. Thus the failures in the patients may have causes other than prednisolone treatment. PMID- 10665720 TI - Total elbow arthroplasty in rheumatoid arthritis: 20 GSBIII prostheses followed 2 5 years. AB - From 1993 to 1996, we implanted 20 primary GSB IlI prostheses in 17 patients with rheumatoid arthritis. The Mayo Clinic performance index for the elbow was used for the evaluation. The average follow-up was 3 (2-5) years. At the follow-up examination, 12 elbows had an excellent result and 8 a good result. The median performance index increased from 30 (15-53) points to 95 (80-100) points. The subjective assessment was excellent for 11 elbows, good for 8 and poor for 1.2 elbows had radiographic loosening with a progressive radiolucent line and a change in the orientation of the prosthesis. PMID- 10665721 TI - Increased migration of the SHP prosthesis: radiostereometric comparison with the Lubinus SP2 design in 40 cases. AB - 40 patients with primary arthrosis were randomized to receive either a cemented SHP (Scientific Hip Prosthesis) or Lubinus SP2 prosthesis. At 2 years radiostereometric measurements showed increased proximal migration (0.4/0.2 mm; p = 0.02) and more proximal wear (0.3/0.1 mm, p = 0.01) of the SHP socket. The SHP stem also subsided (-O.6/-0.1 mm, p<0.001) and rotated more into retroversion (2.6/0.3 degrees ) than did the SP2 design. This subsidence mainly occurred inside the cement mantle in 17 of 18 cases (13 SHP, 4 SP2), where this type of motion could be measured. The changes in bone mineral density evaluated with DEXA and the clinical results did not differ between the 2 groups. The subsidence of the SHP stem is the most pronounced so far recorded with radiostereometry in stems without a completely polished surface. This subsidence and the rotational instability imply a substantial risk of abrasive wear and increased stresses in the cement mantle. PMID- 10665722 TI - Metal-on-metal bearing in hip prosthesis generates 100-fold less wear debris than metal-on-polyethylene. AB - Aseptic loosening due to osteolysis in total hip replacement has been related to wear debris released from prosthetic components. Retrospective longterm observations of patients with the metal-on-metal prosthesis has shown long-term survivorship and good mechanical performance. Thus, the new and modified metal-on metal prosthesis has been introduced on the market. Historical clinical data from the 1st generation metal-on-metal hip prosthesis may not be relevant for the 2nd generation of metal-on-metal hip prosthesis. Therefore, preclinical testing of the prosthesis must be conducted before clinical evaluation. We assessed the tribological performance of the metal-on-metal prosthesis versus the metal-on polyethylene prosthesis introduced on the market as Metasul and Protasul, respectively. In a 12-channel joint simulator, 6 metal-on-metal bearing and 3 metal on polyethylene prostheses were tested, with the same number of corresponding soak controls. The wear was assessed gravimetrically. The "steady state" wear-rates from the metal-on-metal prosthesis were almost 100 times less than that from the metal-on-polyethylene prosthesis. The tribological wear performance of the metal-on-metal hip prosthetic system is promising. PMID- 10665723 TI - Acetabular coverage of the femoral head after triple pelvic osteotomy: no relation to outcome in 51 hips followed for 8-15 years. AB - In developmental dysplasia of the hip in adolescents and young adults, pelvic osteotomies aim to improve acetabular coverage of the femoral head by reorienting the acetabulum. We determined whether acetabular coverage is related to long-term clinical results after triple osteotomy of the pelvis. We used a previously published computer program (Konishi and Mieno 1993) which calculates three dimensional coverage of the femoral head from plain anteroposterior radiographs. We studied the pelvic radiographs of 51 hips in 43 patients and the results were correlated with studies on clinical outcome (de Kleuver et al. 1997). Total acetabular coverage improved from a mean of 56% to 70%. We did not find a relationship between total acetabular coverage and long-term outcome, nor could we determine an optimal coverage. Reduced coverage of the posterolateral quadrant of the femoral head was related to a reduced score for walking ability (p = 0.03), and therefore care should be taken not to overcorrect the acetabulum forwards when attempting to improve the deficient anterior coverage. We challenge the concept that total acetabular coverage is a prerequisite for a good long-term outcome after triple pelvic osteotomy, and hypothesize that other factors such as the change in load across the hip are probably more important in determining the outcome. PMID- 10665724 TI - Fluid flow around model femoral components of differing surface finishes: in vitro investigations. AB - We studied fluid flow at the stem-cement interface of bonded and debonded, polished and rough model femoral components. In a first series of experiments, fluid flow along the interface between bone cement and well-fixed model femoral components, differing in surface finish, and in shape, was measured. Fluid migration along the bone-cement interface of rough stems (Ra 3 microm) was greater than that on polished stems (p < 0.001). This was true of cylindrical and conical tapered stems. On stems with the same surface finish, shape did not influence fluid migration. In a second series of experiments, fluid flow along the stem-cement interface of 5 highly polished and 10 rough-finished (5 of Ra approximately 1.5 microm and 5 of Ra approximately 3 microm), debonded, tapered circular stems was measured. None of the rough stems could prevent fluid flow along the stem-cement interface. Polished tapered stems sealed the interface and, after 48 hrs of continuous pressure, no fluid flow was observed. This difference in the ability to seal the stem-cement interface between rough and polished stems was significant (p < 0.001). The difference in fluid migration along the stem cement interface of rough and polished stems which we observed offers a plausible explanation of the occurrence of osteolysis distal to the articulation of cemented THR in the presence of cement mantle defects. It may also explain why osteolysis is uncommon with polished double-tapered stems. PMID- 10665725 TI - Incomplete cement mantles in the sagittal femoral plane: an anatomical explanation. AB - The influence of operative technique on the formation of incomplete cement mantles in the sagittal plane has been rarely considered in the literature. In this article, we discuss the influence of the anatomy of the proximal femur on the formation of incomplete cement mantles and discuss how their incidence can be reduced by correct component positioning. PMID- 10665726 TI - Open reduction of late unreduced traumatic posterior hip dislocation in 12 children. AB - We present 12 children with late unreduced traumatic posterior dislocation of the hip. All had posterior dislocation and no associated fracture. The dislocation had remained unreduced for a mean period of 20 (6-52) weeks. Open reduction was done in all cases, since none of the hips could be reduced with upper tibial skeletal traction in abduction. All the hips showed varying degrees of avascular necrosis, with preservation of joint space on roentgenograms. 11 children had an excellent outcome, according to the criteria of Garrett et al. (1979), after a follow-up of mean 26 (24-36) months. We suggest that open reduction is a satisfactory treatment for hip dislocation of any duration in children. PMID- 10665727 TI - Bone mineral and migratory patterns in uncemented total knee arthroplasties: a randomized 5-year follow-up study of 38 knees. AB - We measured the amount of bone mineral in the medial tibial condyle 1 week postoperatively, after 1 year and after 4-5 years in 38 arthrotic knees randomized to a Freeman-Samuelson hydroxyapatite-coated (FS HA) or a Miller Galante II (MG II) total knee arthroplasty. Clinically excellent results were recorded in both groups after 5 years. At the last follow-up, the overall decrease in bone mineral was 26%, as measured by triple-energy X-ray absorptiometry. The decrease was larger in FS HA knees than in MG II knees after 4-5 years, indicating stress-shielding of the proximal tibia. Radiostereometry at 1 and 5 years showed smaller maximum total point motion, maximum subsidence and varus or valgus tilt in the FS HA group. There was a tendency towards a reversed relationship between subsidence and change in bone mineral after 1 year, but not after 4-5 years. Distal fixation of the stem in the Freeman-Samuelson hydroxyapatite-coated (FS HA) components might explain the more pronounced loss of bone mineral in the medial tibial condyle. PMID- 10665728 TI - Cancer incidence after total knee arthroplasty: a nationwide Finnish cohort from 1980 to 1996 involving 9,444 patients. AB - A nationwide, computer-based survey of all total joint arthroplasties performed in Finland has been carried out since January 1980. From these records, a cohort of 9,444 patients, with 51,756 person-years, after primary operation with a total polyethylene-on-metal knee arthroplasty (TKA) was followed up for cancer through the Finnish Cancer Register up to December 31, 1996. During the follow-up, 706 cancers were observed. The expected number, based on national rates, was 719; therefore, the standardized incidence ratio (SIR) for all cancers was 0.98. The SIRs for non-Hodgkin's lymphoma (1.40), Hodgkin's disease (1.24) and multiple myeloma (1.54) were increased, but only that of non-Hodgkin's lymphoma was statistically significant 3-10 years after the operation. The numbers of observed cases of prostate cancer exceeded that of expected, with a SIR value of 1.49. A low SIR of lung cancer was observed among men, especially during the first 3 years (0.61), but not in women. The SIR for colon cancer was below unity in women only (SIR 0.70). The SIR for cancer of the urinary organs was close to unity (0.97). SIR relating to soft tissue and bone cancer did not differ significantly from unity, and none of the 6 sarcomas was observed at the site of a prosthesis. The overall cancer risk after TKA done for primary osteoarthrosis seems not to be increased. The increases in lymphoma and prostate cancer risk, however, are observations that could be related to TKA and justify further follow-up of the cohort. PMID- 10665729 TI - Tibialis posterior tendon abnormalities in feet with accessory navicular bone and flatfoot. AB - To assess tibialis posterior tendon (TPT) pathology, we investigated 27 feet with the accessory navicular bone and 22 normal feet by MRI. We found two major anatomical differences in the feet with the accessory navicular bone; the TPT directly inserted in the accessory navicular bone, without any continuity to the sole of the foot or with a slip, less than 1 mm in thickness, and there was a mass with the density of fibrocartilage tissue, between the tendon and the bone in 20/27 feet. These abnormalities were not detected in the control group. 3 patients in the study group were operated on and the MRI findings were confirmed. These findings suggest that patients with the accessory navicular bone and flatfoot should be examined by MRI for insertion abnormalities of the TPT. PMID- 10665730 TI - Decreasing incidence of fractures in children: an epidemiological analysis of 1,673 fractures in Malmo, Sweden, 1993-1994. AB - The incidence of fractures in children in the city of Malmo, Sweden, almost doubled between 1950 and 1979. To see whether a further increase had occurred, we carried out an epidemiological analysis of fractures among children 0-16 years in Malmo 1993-1994. During the study period, 1,673 fractures occurred in 1,610 children. The commonest fracture location was the distal forearm (26%), followed by the phalanges of the hand (16%) and the clavicle (9%). The annual fracture incidence was 235/10(4) in boys, 149/10(4) in girls and 193/10(4) for both genders. This means a decrease in the annual fracture incidence by 9% since 1975 1979. The decrease was not associated with any specific type of fracture or etiological factor. Fractures of the distal forearm among girls were an exception to the general decline, having increased by one third since 1975-1979, which might be explained by the fact that today girls participate to a greater extent in the same sports as boys. PMID- 10665731 TI - Synovial chondromatosis of the wrist and hand--a case report. PMID- 10665732 TI - Accidental Salmonella enteritidis vaccine injection leading to finger necrosis--a report of 4 cases. PMID- 10665733 TI - The role of prophylactic pinning in the treatment of slipped capital femoral epiphysis--a case report. PMID- 10665734 TI - Irreducible fracture-separation of the distal ulnar epiphysis in the Galeazzi equivalent fracture--a case report. PMID- 10665735 TI - Tuberculosis of talus and cuboid--a report of 2 children. PMID- 10665736 TI - Classification and diagnosis of seronegative spondyloarthropathies. PMID- 10665737 TI - Classification and diagnosis of seronegative spondylarthropathies--comments. PMID- 10665738 TI - The prevalence of rheumatoid arthritis in Sweden. AB - The aim of this study was to ascertain the prevalence of rheumatoid arthritis (RA) in a Swedish general adult population. A questionnaire about chronic pain was mailed to a total of 3928 subjects who were chosen as a random sample of the population in two communities in the county of Halland. All persons answering affirmatively to questions intended to identify patients with RA were invited to a clinical examination. X-rays of hands and feet, and analyses of rheumatoid factor and C reactive protein were performed provided that the patients fulfilled two or more of the five clinical items of the 1987 ARA criteria. Furthermore, non participants were searched for in a patient register and in medical records from the local rheumatology unit in an attempt to identify further cases. Using the modified 1987 ARA criteria for population studies the prevalence rate of RA was calculated to 0.51% (95%, CI = 0.31-0.79). PMID- 10665739 TI - Anticardiolipin and anti-beta2GPI antibodies in a large series of European patients with systemic lupus erythematosus. Prevalence and clinical associations. European Concerted Action on the Immunogenetics of SLE. AB - OBJECTIVE: To test the prevalences and the clinical associations of anticardiolipin (aCL) and anti-beta2GPI (abeta2GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). METHODS: 574 SLE patients from 7 European countries were tested for aCL and abeta2GPI by ELISA methods. RESULTS: aCL of IgG, IgM, and IgA isotypes were detected in 22.8%, 14%, and 13.9% of the patients, respectively. IgG and IgM abeta2GPI were detected in 20% of the patients. The presence of aCL was highly associated with the presence of abeta2GPI. Medium-high titer IgG aCL and abeta2GPI were associated with thrombosis, with similar sensitivity, specificity, and positive predictive value. When present at medium-high titer, IgG aCL were associated with thrombocytopenia, IgM aCL with hemolytic anemia, and cerebrovascular accidents. IgA aCL with livedo reticularis and Raynaud's phenomenon. CONCLUSIONS: aCL, when present at medium high titer, are as important as abeta2GPI, as a risk factor for thrombosis. Medium-high titer aCL, but not abeta2GPI, are associated with other clinical features of the antiphospholipid syndrome. PMID- 10665740 TI - Perinatal outcome in pregnancies of women with connective tissue disease and inflammatory rheumatic disease in Norway. AB - Perinatal outcome in infants of women with rheumatic disease notified between 1967 95 to the Medical Birth Registry of Norway was compared to women without such disease. Logistic regression provided odds ratios for associations between rheumatic disease and perinatal outcome for 3 time periods: 1967-76, 1977-86, and 1987-95. Women with rheumatic disease had significantly higher rates of preterm birth than references and this was only partly correlated to the increased occurrence of preeclampsia. The risk of small for gestational age (SGA) infants was significantly higher both in women with connective tissue disease (CTD) and inflammatory arthritides. The proportion of infants with Apgar score < = 6 after 1 minute and 5 minutes was significantly increased in the CTD group indicating moderate to severe fetal asfyxia. The rate of perinatal mortality was high in the CTD group and postperinatal mortality was increased in infants born to mothers with rheumatic disease. Thus, rheumatic disease not only comprises pregnancy outcome, but increases the risk of adverse perinatal outcome. PMID- 10665741 TI - Do flares of systemic lupus erythematosus decline after menopause? AB - OBJECTIVE: To study whether flares of SLE decline after menopause. METHOD: 34 postmenopausal SLE patients with premenopausal disease onset were studied. The frequency and severity of flares before and after menopause was compared. 17 postmenopausal onset SLE patients were also included for comparison. RESULT: Flares in postmenopausal SLE patients decreased significantly after menopause (total No. of flares/patient-year before and after menopause were 0.50+/-0.10 and 0.14+/-0.05, respectively, p = 0.002). The frequency and proportion of severe flares also dropped significantly. The rate and magnitude of postmenopausal flares in these patients were similar to those of the postmenopausal onset SLE patients, a subset known to run a more benign course. CONCLUSIONS: SLE flares less frequently and seriously after menopause. While this may suggest a protective role of hypoestrogenemia against lupus flares, the contribution of other factors like disease duration and effective treatment to this postmenopausal decline of flares cannot be separated from menopause per se. Further studies are needed. PMID- 10665742 TI - Bone mineral density of the lumbar spine in patients with advanced rheumatoid arthritis. Influence of functional capacity and corticosteroid use. AB - We investigated factors that are related to generalized osteoporosis in advanced rheumatoid arthritis (RA). In this cross-sectional study we measured trabecular bone mineral density (BMD), by quantitative computerized tomography (QCT), in the lumbar spine of 57 patients with RA, most of whom were premenopausal women. In our material, 27 out of 57 patients (47%) had BMD <-1 SD expressed as Z-score and five patients had suffered from fractures. Our study shows that a cumulative corticosteroid dose (r = -0.41, p<0.010) and functional impairment (r = -0.37, p<0.050) were negatively related to spinal BMD, while daily intake of calcium correlated positively on BMD (r = 0.37, p<0.010). Our results indicate that low BMD is common in patients with advanced RA and it is associated with long-term corticosteroid use. Thus, in clinical practice we have to consider the benefits and harms of corticosteroid treatment and preventive therapy to osteoporosis. PMID- 10665743 TI - Prolactin, growth hormone, and IGF-1 in ankles and plasma of adjuvant arthritic rats. AB - In this study we have investigated the levels of prolactin, growth hormone, and insulin-like growth factor-1 in plasma and in tissue extracts of ankle joints of rats with acute or chronic adjuvant arthritis using enzyme immunoassay (EIA) and radioimmunoassay (RIA). We found a stable content of prolactin in plasma of the different groups but a significantly increased concentration of growth hormone was observed in the plasma of the group with chronic arthritis. Moreover, an increased concentration of insulin-like growth factor-1 was noted in the plasma of the acute group. This evidently had returned to normal levels in the chronic group. In contrast, decreased concentrations of prolactin, growth hormone, and insulin-like growth factor-1 were found in tissue extracts of ankle joints of the group with chronic arthritis. The changes in the levels of these hormones in adjuvant arthritis might suggest that they play a role in the pathogenesis of the disease. Understanding the mechanism(s) of hormonal participation in adjuvant arthritis may open new treatment strategies for rheumatoid arthritis and other inflammatory disorders. PMID- 10665744 TI - Color duplex ultrasonography in large-vessel giant cell arteritis. AB - In a patient with active extracranial giant cell arteritis, duplex ultrasonography demonstrated hypoechoic mural thickening of the brachial, axillary, subclavian, and carotid arteries with bilateral subtotal occlusions of the brachial and axillary arteries. The ultrasound image of the artery walls became midechoic within 8 weeks, and hyperechoic within one year after start of treatment with corticosteroids. A similar hypoechoic mural thickening of the temporal arteries has been recently described in active giant cell arteritis. The dark ultrasound image is due to an edema of the vessel wall in the acute stage. The brighter ultrasound image might be due to fibrosis in the chronic stage of the disease. PMID- 10665745 TI - AV-block III in a patient with sarcoidosis mimicking Sjogren's syndrome. AB - A 55-year old woman with a diagnosis of primary Sjogren's Syndrome suddenly developed AV-block III. A diagnostic procedure finally revealed sarcoidosis with multiorgan involvement. PMID- 10665746 TI - Tuberculosis of skeletal muscle in a case of polymyositis. AB - We describe a patient with polymyositis receiving corticosteroid therapy, who presented with persistent fever and mass lesion at the left thigh. Surgical exploration and mycobacterial culture proved to be Mycobacterium Tuberculosis infection involving the semitendinous muscle of the left thigh. Suitable surgical debridement, anti-TB medications, and sufficient corticosteroid administration resulted in a good control of both polymyositis and the tuberculous infection. PMID- 10665748 TI - Correction for covariates. PMID- 10665747 TI - Elevated plasma levels of transforming growth factor beta 1 in patients with aseptic loosening of total hip arthroplasties. PMID- 10665749 TI - Is there a role for fractionated radiotherapy in the treatment of arteriovenous malformations? AB - Single fraction radiotherapy for the treatment of arteriovenous malformations (AVMs) is appropriate when brain tolerance is not a limiting factor, but when tolerance is a concern, there is a potential for therapeutic gain with fractionated treatment. alpha/beta values for AVM obliteration have been derived and found to be higher than previously assumed and are likely to be approximately 10.0 Gy or more compared with approximately 1.5 Gy for the tolerance of small volumes of brain. Models are derived to describe, qualitatively, the potential gain that can be achieved with fractionation. Past experience has identified an important volume effect that has limited the use of stereotactic treatments to small volumes. Volume-dependent, dose-volume relationships are described, including a tissue-specific volume exponent (phi) which was found to apply across a relatively wide range of target volumes, thereby permitting the derivation of tolerance guidelines to volumes larger than previously available. More data to define parameter values more precisely are desirable. PMID- 10665750 TI - Smoking and survival from lung cancer. AB - The role of smoking in the etiology of lung cancer is well known but less is known about whether smoking also affects the pathological process. In the present analysis 290 lung cancer patients were divided into three equal-sized groups according to lifetime cigarette consumption (heavy, moderate, light/none). The age of the patient, histology, TNM and S-phase fraction of the tumor were confirmed as prognostic factors. The association between smoking and these factors was evaluated. There was a significant difference in histological type of tumor; adenocarcinomas were more common than other types among light smokers. Tumor size, nodal status and occurrence of distant metastases did not differ significantly between the smoking groups. Heavy smokers had a greater frequency of aneuploid tumors and the S-phase fraction was higher among these patients but neither of these differences was statistically significant. Survival analysis indicated no statistically significant association between survival and lifetime total cigarette consumption. Apart from histological type, smoking is not materially associated with clinical and biological prognostic factors, nor does smoking materially affect the survival of lung cancer patients. Our study material was relatively small and therefore not of sufficient power to detect small effects. PMID- 10665751 TI - Metastatic pattern in non-resectable non-small cell lung cancer. AB - This study describes the metastatic pattern at autopsy in patients with non resectable non-small cell lung cancer (NSCLC) and evaluates the impact of various pretreatment variables and treatment outcomes on the metastatic spread. In eight phase II chemotherapy trials from 1985 through 1993, 337 patients were treated and 51 autopsies were performed (autopsy rate 15%). The male/female ratio was 31/20, median age 56 years (range 36-71), response rate to chemotherapy 8%, and median survival 88 days (range 3-899). Histologic types included adenocarcinoma, 31 cases (60%), squamous cell carcinoma, 9 cases (18%), large cell carcinoma, 9 cases (18%), and unclassified NSCLC, 2 cases (4%). Patients who were autopsied had a shorter median survival than patients without autopsy (p = 0.002, log-rank test). Most commonly involved metastatic sites found at autopsy were mediastinal lymph nodes (84%), pleura (51%), liver (47%), bone (34%), brain (32%), pericardium (29%), adrenals (29%). The median number of involved organs was 5 (range 1-16), with a median of 3 intrathoracic sites (range 1-8) and 2 extrathoracic sites (range 0-11). Patients who initially had metastatic NSCLC also had significantly more metastatic sites at autopsy both extrathoracic (p = 0.004) and totally (p = 0.03) compared to patients with locally advanced disease. No other relation to pretreatment variables was found. PMID- 10665752 TI - Endobronchial radiotherapy for malignant bronchial obstruction or recurrence. AB - Forty-five lung cancer patients who had recurrence and/or endobronchial obstruction were treated with intrabronchial radiotherapy (IBRT). The majority had been heavily treated previously, mainly by external radiotherapy; six patients were treated surgically. IBRT was given with high-dose-rate equipment, either 18 Gy in three fractions or 15 Gy as a single dose was originally planned. For different reasons several patients received other regimens. Palliation of dyspnoea was obtained in 64% of the patients. Palliation of hemoptysis (12/14) and cough (11/17) was registered, as well as improvement in atelectasis in 11/26 patients. Palliation of dyspnoea was enhanced with an IBRT dose > 15 Gy. The median survival after IBRT was 13 weeks. Fatal hemoptysis (FH) occurred in 12 patients; 10 of these within 6 months after treatment. The risk of FH significantly increased with an IBRT dose > 15 Gy. PMID- 10665753 TI - Induction chemotherapy followed by concurrent chemoradiotherapy in stage III unresectable non-small cell lung cancer. AB - The favourable experience with the combination regimen of vinorelbine, ifosfamide and cisplatin (NIP) in patients with metastatic non-small cell lung cancer (NSCLC) has led to a protocol assessing this regimen as an induction treatment in patients with stage III unresectable NSCLC, followed by thoracic radiotherapy with concurrent daily cisplatin as a radiosensitizer. Two cycles of NIP were administered 21 days apart; each cycle comprised i.v. vinorelbine 25 mg/m2 on days 1 and 8, i.v. ifosfamide 3 g/m2 on day 1 with MESNA as uroprotection, and i.v. cisplatin 50 mg/m2 on day 1. Radical thoracic radiotherapy commenced on day 43 to a total dose of 64 Gy and i.v. cisplatin 6 mg/m2 was given concurrently prior to each fraction of radiation as a sensitiser. Two more cycles of NIP were given to patients who responded favourably to the induction treatment about 2 weeks after completion of radiation. Between July 1995 and July 1997, 44 patients were treated with this protocol. This treatment schedule was generally well tolerated. Grade 3-4 neutropenia occurred in 50% of the patients and neutropenic sepsis was seen in 8. Grade 3-4 oesophagitis was uncommon. Most of the patients were able to complete the induction and concurrent chemoradiotherapy phase. Major response occurred in 75% of the patients with 2 (4.5%) complete responses (CR). A total of 6 patients achieved CR after chemoradiotherapy. At a median follow-up of 35 months, the median overall survival for all patients was 15 months with a 3 year survival rate of 24%. The median overall survival for stage IIIA patients was 19 months with a 3-year survival rate of 39% in contrast to 13 months' median overall survival and only 15% 3-year survival rate for stage IIIB. The NIP regimen results in a high response rate in NSCLC and this treatment programme seems to benefit selected patients with stage III disease. PMID- 10665754 TI - Factors influencing the distribution of metastases and survival in extensive disease small cell lung cancer. AB - This study was conducted to investigate the distribution of metastatic lesions and their influence on survival, as well as other prognostic factors previously shown to have an impact on the outcome of patients with extensive small cell lung cancer (SCLC). Of the 207 patients were included and retrospectively analyzed; 124 patients had extended disease at initial presentation and the remaining 83 developed metastatic disease during follow-up. Patients who relapsed presented most frequently with distant metastases. The brain was the most frequent organ targeted for metastatic disease following the completion of chemotherapy (p<0.05). Serum LDH levels correlated significantly with the presence of liver metastasis (p<0.001). The site of involvement did not seem to have an impact on survival. Nevertheless, patients with multiple metastatic sites had a significantly poor survival rate (p = 0.001). Weight loss, performance status, gender, clinical stage, serum LDH and albumin levels were all shown to correlate with survival (p<0.05). Response to chemotherapy was determined to be the most important prognostic factor. PMID- 10665755 TI - Vincristine (Vinca-alkaloid) as a sclerosing agent for malignant pleural effusions. AB - Vincristine, extracted from Vinca rosea Linn., is an effective antineoplastic chemotherapeutic drug used in oncology practice. This drug has never been used as a sclerosing agent for the treatment of malignant pleural effusion for reasons unknown. A study was conducted to examine the use of Vinca-Alkaloid as a sclerosing agent (pleurodesis) for the palliative treatment of malignant pleural effusions. The study included 15 patients, all diagnosed to have cytology-proven malignant pleural effusions. Intercostal tube drainage followed by chemical sclerotherapy with 2 mg vincristine was performed on all patients and a high success rate was noted. Twelve procedures out of 15 (12/15) achieved complete resolution of pleural fluid with a success rate of 80%. In two procedures the pleural effusion was reduced and then recurred but did not require re-aspiration. One procedure failed and repeated pleural aspiration was required. In this study, with adequate pleural drainage and the proper technique, vincristine was found to be an effective sclerosing agent for malignant pleural effusion. Further randomized trials are necessary in order to establish the role of this drug. PMID- 10665756 TI - Cancer of the mobile tongue in Finland--increasing incidence, but improved survival. AB - A population-based descriptive study was conducted to describe incidence and survival of cancer of the mobile tongue in Finland between 1953 and 1994. The study included 1504 patients, drawn from the Finnish Cancer Registry, with first primary mobile tongue cancer diagnosed between 1953 and 1994. Incidence and relative survival were determined. The age-standardized overall incidence rate was 0.6 per 100000 years in 1953-1994. At the time of diagnosis 78% of the patients had either localized or regional disease. The age-standardized incidence rate decreased after the mid-1960s, but increased in the 1990s. The 5-year relative survival rate increased gradually from 40% in 1953 1959 to 58% in 1988 1994. Disease stage at the time of the diagnosis strongly affected the survival rate. Survival increased especially in regional disease. Cancer of the mobile tongue is increasing in Finland, but survival has increased particularly in regional disease, probably because of improved treatment. Early diagnosis is emphasized for a good prognosis. PMID- 10665757 TI - Tumor volume and local control in primary radiotherapy of nasopharyngeal carcinoma. AB - An investigation of the effect of tumor volume and total dose on local control following primary radiotherapy for nasopharyngeal carcinoma was carried out in order to estimate the radiation dose necessary to control a specific tumor volume. Between 1983 and 1996 a total of 104 patients underwent radiation therapy for nasopharyngeal carcinoma at the Department of Radiation Oncology of the University of Wuerzburg. Total doses of between 8 and 80 Gy (5 fractions per week) were administered. Complete CT-data on primary tumor size, total tumor dose (calculated by 3D- or quasi 3D-CT-based radiation planning computer) and on local control status in the follow-up period were available for 63 patients. Lymph node metastases were present in 38 of these patients and they were also entered into the study. Thus this study is based on a total of 101 tumor regions. A Poisson probability-based model was used for calculating the dose-response relationship. Assuming a correlation between tumor volume and the total dose necessary to obtain local control, the individual tumor volumes were rescaled to a 1 ml volume by introducing a volume-dependent modification factor for the applied dose, in order to eliminate the influence of different individual tumor volumes. All dose values given are based on a fractionation scheme of 2 Gy single dose, 5 fractions per week. Nineteen tumors and 11 lymph nodes were considered locally uncontrolled or recurrent. Without dose-volume modification, a weak dose-response correlation was found and a typical shallow dose-response curve was calculated with a 50% response dose (RD50) of 60.2 Gy and a normalized dose-response gradient (gamma50) of 3.2+/-0.62. After dose-volume modification and rescaling to a 1 ml tumor volume, a steep dose-response curve with an RD50 of 40.9 Gy and gamma50 of 8.2. was found. Tumor volume is a very important factor influencing local control in nasopharyngeal carcinoma. The rescaling procedure to a reference volume of 1 ml used in this study revealed a very steep dose-response relationship. This result suggests that the clinically observed smooth dose-response relationships may be explained by interindividual tumor volume heterogeneity. The additional dose necessary to control a tumor of the double volume is close to 5 Gy. With a total dose of 72 Gy (5x2 Gy/week), tumor volumes larger than 64 ml are unlikely to be controlled. PMID- 10665758 TI - Radiation therapy and concurrent cisplatin in management of locoregionally advanced nasopharyngeal carcinomas. AB - Radiation therapy in combination with chemotherapy in the management of locoregionally advanced nasopharyngeal carcinomas is evaluated in an attempt to improve locoregional response, reduce locoregional failure and reduce systemic failure. The current study was designed to investigate radiation therapy and concurrent cisplatin in this context. From 1992 through 1997, 70 patients with locoregionally advanced nasopharyngeal carcinomas were treated with radiation therapy and concurrent cisplatin. External beam radiation dose was 60 Gy for T1, T2 and T3 tumors, 70 Gy for T4 tumors and 70 Gy for metastatic cervical lymph nodes. An intracavitary brachytherapy boost (10 Gy) was applied for T1, T2 and T3 tumors. Cisplatin (30 mg/m2) was administered weekly during external beam radiation therapy. Locoregional complete response was achieved in 63 patients, locoregional failure was observed in 4 patients and systemic failure was observed in 15. N-stage predicted systemic failure. Overall survival, locoregional failure free survival and systemic failure-free survival were 63%, 79% and 75%, respectively, at three years. Grade 3 acute skin toxicity was observed in 2 patients, Grade 3 acute mucous membrane toxicity was observed in 6 and Grade 3 acute hematological toxicity was observed in 2 patients. Despite improved locoregional response, reduced locoregional failure and improved survival with radiation therapy and concurrent cisplatin, systemic failure remains prevalent for locoregionally advanced nasopharyngeal carcinomas. PMID- 10665759 TI - Changes in oxygen tension during radiotherapy of head and neck tumours. AB - Increased knowledge about changes that occur in tumour oxygenation during radiotherapy and the biological factors causing these changes can be useful in the development of optimal radiation treatments. The aims of this study were a) to study changes in the oxygen tension (pO2) of human head and neck tumours during radiotherapy in relationship to changes in cell density and vascular density, and b) to investigate whether the pO2, measured before or during therapy, can be used to predict the therapeutic outcome. Preliminary data from the first 11 patients included in the study are reported. The pO2 was measured before treatment (11 patients) and once a week during therapy (8 patients), using polarographic needle electrodes. Cell density and vascular density were determined from biopsies taken after each pO2 measurement in 5 patients. Significant fluctuations in pO2 occurred during therapy. Changes in hypoxic fraction; i.e., fraction of pO2 readings below 2.5 mm Hg, 5 mm Hg or 10 mm Hg, coincided with changes in cell density, but not with changes in vascular density, which suggests that the changes in hypoxic fraction were caused by changes in oxygen consumption rather than supply. Response evaluation after a median follow up time of 19 months showed that progressive disease occurred among the patients with highly hypoxic tumour, regardless of whether hypoxic fraction before treatment or after two weeks of radiotherapy was considered. PMID- 10665760 TI - Long-term continuous 5-fluorouracil infusion in patients with advanced head and neck cancer. AB - Forty-two patients with advanced head and neck cancer entered this phase II trial of long-term continuous 5-fluorouracil (5-FU) infusion at a dose of 300 mg/m2/day for a maximum of 16 weeks. Objective response rate was 15% in 41 evaluable patients. Median time to progression was 2.9 months, and median survival 4 months. Toxicity was generally mild. Reversible stomatic and hand-foot syndrome WHO grade III-IV was observed in 5 and 3 patients, respectively. Haematologic toxicity and emesis were less pronounced with no grade III-IV toxicity. One patient had to discontinue treatment because of ataxia. No catheter-related toxicity and no treatment-related mortality were observed. In the present study long-term continuous infusion of 5-FU has only modest activity in terms of response rate, but the activity is comparable with other single-agent regimens. The treatment is well tolerated, with minimal toxicity making it usable in a palliative situation. PMID- 10665761 TI - Alterations in head and neck cancer occurring in HIV-infected patients--results of a pilot, longitudinal, prospective study. AB - To assess the impact of human immunodeficiency virus (HIV) infection on the presentation and course of head and neck squamous cell carcinoma (HNSCC), we performed a pilot, prospective, longitudinal study of all patients with HNSCC presenting to our institutions over a 6-month period (n = 10). A 60% incidence of HIV infection was seen in this study population, with SCC presenting as the initial manifestation of HIV infection in 2 of the 6 patients. In addition. HIV infected patients were significantly younger than non-infected patients at (p = 0.01). None of the HIV-infected patients had acquired immunodeficiency syndrome (AIDS) at the time of presentation, but 5 of 6 patients had an abnormal CD4 count, compared to none of the non-infected patients (p = 0.05). The absolute CD4 count in HIV-infected patients decreased to less than 100x10(9)/L in the majority of these patients within 3 months of presentation with HNSCC (p = 0.05). Treatment-associated complications were common in HIV-infected patients, occurring in 4 of the 6 cases in contrast to none of the patients without HIV infection (p = 0.046). Outcome was significantly poorer for HIV-Infected patients, with 5 patients succumbing to their disease within one year, in contrast to none of the non-infected patients (p = 0.046). These data, combined with our previous work, justify further investigation of the relationship between HNSCC and HIV infection and the possibility of its inclusion as an AIDS-defining process. PMID- 10665762 TI - Low-grade astrocytoma--a retrospective analysis of 102 patients. AB - One hundred and two patients (57 males, 45 females, median age 17 years) with histologically proven low-grade astrocytoma (grades I, II) treated between 1978 and 1994 were retrospectively analyzed at the King Faisal Specialist Hospital & Research Center. Microscopic investigation showed 50 patients (48%) with grade I tumors as opposed to 52 patients (52%) with grade II tumors. Fifteen patients (15%) had complete surgical excision, 55 (52%) had partial excision and 32 (31%) had biopsy only; 68 patients (66%) received external radiotherapy with a median dose of 54 Gy (range 45-68.5 Gy). With a median follow-up of 3.3 years, the 5 and 10 years, overall actuarial survival rates were 78% and 62%, respectively while the progression-free survival rates at 5 and 10 years were 69%, and 35%, respectively. Age and performance status were significant prognostic factors in terms of overall survival on univariate (p = 0.05 and 0.05, respectively) and multivariate analysis (p = 0.005 and 0.006, respectively). PMID- 10665763 TI - Prognostic significance of the PC10 index for patients with stage II and III oesophageal cancer treated with radiotherapy. AB - The monoclonal antibody PC10 is used for immunohistochemical staining of the proliferating cell nuclear antigen (PCNA). The percentage of PC10-positive cancer cells is defined as the PC10 index. We evaluated the relationship between the PC10 index in pretreatment endoscopic biopsies and the prognoses of 47 patients with Stage II-III oesophageal squamous cell carcinoma treated with radiotherapy. The patients with a PC10 index > 40% had significantly poorer prognoses than the other patients (p = 0.0007). Proportional hazards model analysis indicated that only the PC10 index was a prognostic factor (p = 0.0009). The patient group of complete responders showed significantly lower PC10 indices compared to patients with a partial response or no change (p = 0.049). The PC10 index can be a good predictive indicator of the prognosis in patients with Stage II-III oesophageal cancer treated with radiotherapy. PMID- 10665764 TI - Cost-effectiveness analysis of pegylated-liposomal doxorubicin and liposomal daunorubicin treatments in patients with Kaposi's sarcoma. AB - Economic evaluations of new AIDS treatment drugs are important. For physicians treating patients with Kaposi's sarcoma, these issues are especially meaningful since cancer treatment costs for this group of patients are high. Kaposi's sarcoma is the most frequently occurring neoplasm in AIDS patients, affecting about 15% of this population. In our study, a retrospective economic evaluation has been made based on data from two randomized phase III clinical studies of severely immune-compromised HIV-infected individuals and which compares liposomal doxorubicin with liposomal daunorubicin. We have estimated the cost and cost effectiveness of the two drugs. The costs per complete or partial response are USS 18340 for daunorubicin and USS 8871 for doxorubicin. The incremental cost per additional responder by using liposomal doxorubicin instead of liposomal daunorubicin is USS 1910. Sensitivity analysis shows that these results hold over a wide range of assumptions. PMID- 10665765 TI - Epidemiological characteristics of cutaneous malignant melanoma of the head and neck--a population-based study. AB - Since cutaneous malignant melanoma (CMM) and melanoma in situ (MIS) of the head and neck have only partially been differentiated from CMM of other anatomic sites, these lesions are classified in detail in this study. Data from 756 patients derived from the population-based register of the Stockholm-Gotland area were analyzed and the findings showed that the incidence of CMM was 3.4 times higher in the face compared to the skin outside the head-neck area and that lentigo maligna melanoma was 74 times and nodular melanoma 2.3 times more common in the face. Mean age at diagnosis was significantly higher for patients with CMM of the head and neck irrespective of histogenetic type. Tumor site within the head and neck related to age at diagnosis. CMM of the head and neck differs from CMM of other locations. Epidemiological data are in agreement with the hypothesis that UV radiation (chronic or intermittent) may give rise to melanomas with various phenotypic traits. PMID- 10665766 TI - Radioimmunotherapy of DU-145 tumours in nude mice--a pilot study with E4, a novel monoclonal antibody against prostate cancer. AB - The anti-tumour effect of the 131I-labelled antiprostate monoclonal antibody (MAb) E4 was studied in an experimental model with 41 nude mice, subcutaneously xenografted with a human prostate cancer cell line (DU-145). The mice were divided into four study groups, i.e. one receiving single and another repeated injections of the radiolabelled MAb. A third group was injected with non-labelled MAb, and the fourth served as an untreated control group. The tumour volumes increased similarly in all groups during the 27-day observation period. The tumour tissue was morphologically disintegrated in the group that received repeated radioimmunotherapy (RIT). The tumours from this group contained large fluid-filled cystic parts and demonstrated pronounced cellular and subcellular polymorphism in the remaining viable tumour tissue. The untreated control tumours and single therapy tumours remained solid. The proportion of the total tumour volume that consisted of viable tumour cells, as determined by morphometric techniques, was significantly lower in the 131I-E4-treated groups. The use of 131I-labelled E4 MAb has thus demonstrated a promising therapeutic potential. PMID- 10665767 TI - Short- and long-term effects of irradiation on laryngeal mucosa of the rat. AB - Although radiotherapy is often used to treat laryngeal carcinoma, there is little information on the effects of this treatment on laryngeal structures. Rats were irradiated to the head and neck region and the larynges were studied by light- and electron-microscopy and immunohistochemistry. Ten days after irradiation, a change in the ultrastructural appearance of the granules of the subglottic glands was observed. Substance P-, bombesin- and enkephalin-like immunoreactivity was increased in local ganglionic cells and glandular nerve fibres. The mast cells were reduced in number. At examination 4 6 months after irradiation, there were no obvious differences compared with controls concerning mast-cell numbers and neuropeptide expression. The ultrastructural changes seen in the subglottic glands remained to some extent. The results show that structural changes in the subglottic glands occur concomitantly with an increased expression of certain neuropeptides in the innervation of these glands, which implies a relationship between these two parameters. The mast cells respond drastically to irradiation, but in the long run, regeneration of these cells occurs. PMID- 10665768 TI - Effects of retroviral-mediated herpes simplex virus thymidine kinase gene transfer to murine neuroblastoma cell lines in vitro and in vivo. AB - Selective introduction of genes conferring chemosensitivity on proliferating tumor cells can be used to treat cancer. We investigated the efficacy of retrovirus-mediated gene transfer of the herpes simplex virus thymidine kinase (HSV-TK) gene to murine neuroblastoma cell lines (neuro-2a) in vitro and in vivo. Retrovirus-mediated HSV-TK gene transfer to the neuro-2a cells resulted in sensitivity to ganciclovir (GCV) in vitro. In A/J mice, tumors produced from HSV TK transduced neuro-2a cells regressed after GCV treatment. Intratumoral injection of recombinant retrovirus expressing HSV-TK gene also inhibited growth of the tumor established in A/J mice. These results demonstrate that HSV-TK gene therapy might be a feasible approach for inhibiting the growth of neuroblastoma. PMID- 10665769 TI - Does bombesin-like peptide mediate radiation-induced anorexia and satiety? AB - Bombesin (BN) and its mammalian counterpart gastrin-releasing peptide (GRP) act as neuroregulatory hormones and peripheral and central satiety-inducing agents. Previously, we demonstrated that irradiation induces an increase in the expression of BN/GRP in the innervation of the salivary glands in rats. We therefore carried out a study using radioimmunoassay (RIA) analysis and immunohistochemistry to examine whether saliva contains BN and whether irradiation affects the BN release to saliva in rats. Immunoreactivity for BN was detected not only in the innervation of the parenchyma but also in the duct cells and in the lumina of the ducts, suggesting entrance of BN into saliva. The RIA analysis confirmed that rat saliva contains a BN-like peptide. The observation shows that saliva contains this peptide but that there is no significant increase following the radiation schedule used. Nevertheless, the occurrence of an enhanced expression of BN in different peripheral tissues such as the salivary and laryngeal glands should be taken into consideration when discussing the clinically important problem of reduced food intake and anorexia in cancer patients. PMID- 10665770 TI - A retrospective analysis of Hickman line-associated complications in patients with solid tumours undergoing infusional chemotherapy. AB - We present a retrospective analysis of Hickman line use and associated complications in patients with solid tumours undergoing treatment with infusional chemotherapy. One hundred and ten lines were inserted in 94 patients (55 females and 39 males, median age 51), of whom 107 were placed under radiological screening, the remainder by a surgical approach. Catheters were in situ for a total of 9670 days (median 101 days, range 1-278). Fifty-five complications occurred during the lifespan of 39 catheters (35.5%), giving an overall complication rate of 4.03/1000 catheter days. Early complications included pneumothorax (4%), arterial puncture (1%) and failure of placement (1%). Late complications included sepsis (superficial and systemic) (24.5%), venous thrombosis (9%), line displacement (10%) and catheter blockage (1%). Fifteen episodes of systemic sepsis occurred in 12 patients, giving an overall sepsis rate of 1.55/1000 catheter days, while complications requiring catheter removal occurred in 20 cases (18% of insertions, 2.07/1000 catheter days). We conclude that the use of Hickman catheters as a means of long-term venous access in infusional chemotherapy patients is generally safe, but is associated with significant morbidity. PMID- 10665771 TI - Patient positioning using artificial intelligence neural networks, trained magnetic field sensors and magnetic implants. AB - The purpose of this study was to evaluate the precision of a sensor and to ascertain the maximum distance between the sensor and the magnet, in a magnetic positioning system for external beam radiotherapy using a trained artificial intelligence neural network for position determination. Magnetic positioning for radiotherapy, previously described by Lennernas and Nilsson, is a functional technique, but it is time consuming. The sensors are large and the distance between the sensor and the magnetic implant is limited to short distances. This paper presents a new technique for positioning, using an artificial intelligence neural network, which was trained to position the magnetic implant with at least 0.5 mm resolution in X and Y dimensions. The possibility of using the system for determination in the Z dimension, that is the distance between the magnet and the sensor, was also investigated. After training, this system positioned the magnet with a mean error of maximum 0.15 mm in all dimensions and up to 13 mm from the sensor. Of 400 test positions, 8 determinations had an error larger than 0.5 mm, maximum 0.55 mm. A position was determined in approximately 0.01 s. PMID- 10665772 TI - Malignant teratoma of the lung. PMID- 10665773 TI - Microscopic pulmonary tumoral embolism and subacute cor pulmonale as the first clinical signs of cancer. PMID- 10665774 TI - Mucin-negative pseudomesotheliomatous adenocarcinoma of the lung: report of three cases. PMID- 10665775 TI - The immunostimulating activities of anti-tumor polysaccharide from K1 capsular (polysaccharide) antigen isolated from Klebsiella pneumoniae. AB - We have previously shown that K1 capsular polysaccharide antigen (K1CPS) of Klebsiella exhibits anti-tumor activities. In the present study, we examined the effect of K1CPS on cytotoxic effector cells. We found that K1CPS could activate many cytotoxic effector cells including alloreactive cytotoxic T cells and tumor infiltrating lymphocytes (TILs). Moreover, K1CPS could increase the anti-tumor activity of lymphokine-activated killer (LAK) cells, both in vitro and in vivo. The i.p. injection of K1CPS in low dose could enhance the LAK cytotoxicity and the effect was further potentiated by coculture of LAK cells with K1CPS and low concentration of murine rIL-2 in vitro. The phenotypic characterization revealed that K1CPS might contribute to the increase in CD3+ LAK cell subpopulation by its in vivo priming effect. In addition, the K1CPS-treated LAK cells were able to inhibit the growth of WEHI-164 tumor cells in vivo in Winn-type inhibition assay. Subcutaneous (s.c.) and intraperitoneal (i.p.) adoptive infusion of LAK cells (splenocytes from K1CPS-treated WEHI-164-bearing mice cultured with K1CPS-plus rIL-2) into WEHI-164 sarcoma-bearing mice could slightly cause regression in terms of tumor diameter, and more significantly in sarcoma weight. PMID- 10665776 TI - Minor role of the C3a receptor in systemic anaphylaxis in the guinea pig. AB - Previously, Regal et al. [Regal, J.F., Fraser, D.G., Toth, C.A., 1993. Role of the complement system in antigen-induced bronchoconstriction and changes in blood pressure in the guinea pig. J. Pharmacol. Exp. Ther. 267, 979-988] demonstrated that preventing complement system activation resulted in inhibition of anaphylaxis in the guinea pig, and that the C-terminal 21 amino acids of guinea pig C3a (C3a-peptide) mimic the symptoms of anaphylactic shock in the guinea pig [Regal, J.F., 1997. Role of the complement system in pulmonary disorders. Immunopharmacology 38, 17-25]. To determine if C3a is an essential mediator of systemic anaphylaxis, the anaphylactic response to ovalbumin (OA) was assessed in guinea pigs genetically deficient in the C3a receptor (C3aR-) compared to their control strain of animals which were C3a receptor positive (C3aR+). In addition, the response to another control strain of animals, Hartley guinea pigs, was determined. Sensitized guinea pigs were anesthetized, and bronchoconstriction and changes in blood pressure were monitored in response to intravenous (i.v.) injection of either C3a-peptide, recombinant human C5a (rHuC5a) or OA. Both Hartley guinea pigs and C3aR+ animals responded similarly to C3a-peptide and rHuC5a. C3aR- animals, however, were unresponsive to C3a-peptide and responded normally to rHuC5a, confirming their functional deficiency of the C3a receptor. In response to OA, C3aR+ animals and Hartley guinea pigs responded with a severe bronchoconstriction, an initial transient hypotension, followed by an increase in blood pressure and a delayed prolonged hypotensive response. In contrast, in C3aR animals, the increased blood pressure response to OA was significantly prolonged, the delayed hypotensive response was blunted, and the bronchoconstriction was delayed compared to the C3aR+ animals. The difference in the anaphylactic response could not be explained by differing amounts of OA specific IgG1 antibody or C3a generated during the anaphylactic response. Thus, these data suggest that C3a plays a minor role in the hypotension of systemic anaphylaxis and investigation of a role for other products of complement system activation, either alone or in combination with C3a, is clearly warranted. PMID- 10665777 TI - Induction of apoptosis of activated murine splenic T cells by cycloprodigiosin hydrochloride, a novel immunosuppressant. AB - Two types of immunosuppressants, cycloprodigiosin hydrochloride (cPrG) and L leucyl-L-leucine methyl ester (LeuLeuOMe), both have the ability to selectively inhibit the lysosomal function, and a related compound to cPrG, prodigiosin 25-C, and LeuLeuOMe have been reported to selectively inhibit the T cell function in vitro. We therefore examined the cell-type specificity of cPrG and LeuLeuOMe using murine splenocytes. Concanavalin A (Con A)- and lentil lectin-induced proliferation was suppressed by cPrG more profoundly than lipopolysaccharide induced proliferation. At the optimal concentration, Con A induced the proliferation of both CD4+ and CD8+ cells, whereas at a supra-optimal concentration Con A induced rather selective proliferation of CD8+ cells. Irrespective of the dose of Con A, CD4+ and CD8+ cells were equally affected by cPrG. In contrast, LeuLeuOMe induced the selective loss of CD8+ cells. cPrG enhanced the apoptosis of murine splenocytes and nylon fiber column-purified T cells cultured in the presence of Con A, as shown by the decrease in cell size and/or DNA fragmentation. Overall, this study revealed that the cell-type specificity of cPrG is different from that of LeuLeuOMe, and that the immunosuppression by cPrG is associated with apoptosis. PMID- 10665778 TI - Critical role of Kupffer cell CR3 (CD11b/CD18) in the clearance of IgM-opsonized erythrocytes or soluble beta-glucan. AB - Liver macrophages (Kupffer cells) play a major role in blood clearance of both C3 opsonized immune complexes and therapeutic beta-glucan polysaccharides. Human Kupffer cells express three types of C3-receptors: CR1 (C3b-receptor; CD35), CR3 (iC3b- and beta-glucan-receptor), and CR4 (iC3b-receptor; CD11c/CD18). Studies of isolated macrophages have suggested that CR3 is the major receptor mediating capture of either C3-opsonized erythrocytes (E) or beta-glucans. In this investigation, the organ distribution and function of CR3 in the clearance of IgM opsonized E and soluble CR3-binding polysaccharides were explored in normal vs. CR3-knockout (CR3-KO) mice. Analysis of intravenously (i.v.) injected 125I-anti CR3 showed that the major vascular reservoir of CR3 was the liver, followed by spleen and lungs. By contrast, clearance of 125I-anti-CR1 appeared to be mediated predominantly by splenic B lymphocytes, as only subsets of splenic macrophages or Kupffer cells were found to express CR1. Clearance of IgM-opsonized 51Cr-E occurred rapidly to the livers of normal mice but was nearly absent in CR3-KO mice. Soluble 125I-beta-glucan exhibited rapid clearance to the liver in normal mice, whereas clearance in CR3-KO mice was significantly reduced. In conclusion, Kupffer cell CR3 plays a crucial role in the clearance of both IgM-opsonized E and beta-glucans. PMID- 10665779 TI - Selective augmenting effects of nitric oxide on antigen-specific IgE response in mice. AB - Here, we report the enhancing effects of nitric oxide (NO) on an IgE antibody response in mice. Anti-trinitrophenyl (TNP) IgE production induced in vitro in TNP keyhole limpet hemocyanin (KLH)-primed spleen cells was inhibited by approximately 70% when an NO synthase (NOS) inhibitor, L-N(G)-monomethyl-L arginine, was added at 10(-7)-10(-6) M to the lymphocyte culture. On the other hand, addition of NO-generating agents to the culture resulted in a marked enhancement of the IgE production. In contrast, anti-TNP IgM and IgG1 responses were affected only marginally when the IgE production was either suppressed or augmented by these agents. NO did not directly augment IgE class switching in normal B cells stimulated with lipopolysaccharide and interleukin (IL)-4. NO mediated augmentation of the IgE response is considered to be of a physiological significance because administration of aminoguanidine (AG), an inhibitor of inducible NOS, to immunized mice resulted in a preferential suppression of anti TNP IgE production in vivo. This may be explained by the observation that AG administration increased interferon-gamma expression without changing that of IL 4 in the immunized mice. Taken together, these observations suggest a pathophysiological role of NO in the development of IgE-mediated allergic diseases. PMID- 10665780 TI - Immunological parameters in patients suffering from alcohol-dependence syndrome. AB - Alcohol abuse is a major cause of abnormal liver development and activity. In addition to enzymatic malfunction, alcohol and its metabolites induce changes in the levels of some liver antigens, resulting in immunological disturbance. The purpose of the present study is to correlate the severity of liver function impairment with the length of alcohol abuse, in order to be able to use such tests as indicative of the severity of Alcohol Dependence Syndrome. Thirty-one alcohol abusers were allocated to three groups on the basis of the levels of their liver enzymes, and were tested for a variety of immunological parameters and skin reactions. The data indicate that even though not all immunological values measured differed significantly from the control values, in those that did (granulocytes, lymphocytes, CD4/CD8 ratio, C3, IgG, IgM and some skin positive reactions), the biggest difference was between the healthy volunteers and the group with the longest abuse period. It is suggested that changes in selected immunological parameters in alcohol abusers may indicate the severity of their liver dysfunction. PMID- 10665781 TI - Selective cyclooxygenase-2 (COX-2) inhibitors reduce anti-Mycobacterium antibodies in adjuvant arthritic rats. AB - Adjuvant arthritis, induced by Mycobacterium butyricum, is an experimental immunopathy that shares many features of human rheumatoid arthritis and, as such, is one of the most widely used models for studying the anti-inflammatory activity of compounds. In rats with adjuvant induced arthritis, IgG antibodies to M. butyricum have been detected and autoantigens that cross react with mycobacteria may be involved in the pathogenesis of adjuvant arthritis. In this study, the anti-inflammatory and immunosuppressive activities of two cyclooxygenase-2 selective inhibitors, flosulide and L-745,337, at doses of 0.1, 1 and 5 mg/kg/day, were examined in adjuvant arthritic rats. After 14 days of treatment, a clear dose-dependent inhibition of plantar edema was seen for both flosulide (ID50 lower than 0.1 mg/kg) and L-745,337 (ID50 = 0.4 mg/kg). Plasma levels of IgG anti-M. butyricum antibodies were also decreased by both drugs. In each case the maximal immunosuppressive effect was observed at doses lower than 5 mg/kg. The non-selective COX-2 inhibitor, indomethacin (1 mg/kg) decreased paw edema by 65% and the levels of IgG anti-M. butyricum by 45%. Neither cyclooxygenase selective inhibitors nor indomethacin decreased the delayed hypersensitivity reaction induced by M. butyricum. Thus, in vivo inhibition of COX-2 inhibited articular swelling and also the humoral immune response to Mycobacterium. PMID- 10665782 TI - Differential effects of chronic propranolol treatment on the phenotypic profile of thymocytes from immature and adult rats. AB - To elucidate a putative role of beta-adrenoceptors in the modulation of intrathymic T-cell maturation, the expression of major differentiational antigens (CD4/CD8 and TCR alphabeta) on the thymocytes from both immature (aged 21 day at the beginning of the treatment) and adult (aged 75 days at the beginning of treatment) male rats subjected to a 15-day-long propranolol treatment (0.40 mg/100 g/day, s.c.) was analyzed by two- and one-color flow cytometry, respectively. Rats of matched age injected with saline served as controls. The propranolol treatment in immature but not adult rats caused a significant reduction in both the relative thymus weight and total thymocyte yield. In addition, a significant increase in the percentage of CD4+ 8+ double-positive cells, with a proportional decrease in the relative proportion of CD4+ 8- single positive cells, was found in immature rats. In contrast, a slight but significant decrease in the percentage of CD4+ 8+ cells with a parallel increase in the relative proportion of CD4+ 8- cells was found in adult rats. In both groups of rats, the percentage of TCR alphabeta(total) thymocytes was increased: in immature rats this was due to an increase in the percentage of TCR alphabeta(low) thymocytes, while in the adult rats it reflected a rise in the relative proportion of TCR alphabeta(high) cells. In conclusion, the study revealed that propranolol treatment in both immature and adult rats alters the relative proportion of CD4+ 8+ and CD4+ 8- thymocytes, but in opposite fashion, and the data suggest that this treatment affects distinct fractions within the population of CD4+ 8+ thymocytes with respect to expression of TCR alphabeta. The results also indicate that, regardless of rat sexual maturity, the development of thymocytes towards CD4- 8+ T-cells is relatively insensitive to long-lasting beta adrenoceptor blockade. PMID- 10665783 TI - Ascorbic acid modulates in vitro the function of macrophages from mice with endotoxic shock. AB - The toxic effects of oxygen radicals produced by immune cells can be controlled to certain degree by endogenous antioxidants because of their scavenger action. This control is specially important in a type of immune cell, i.e., the phagocyte, which produces oxygen-free radicals and uses antioxidants in order to support its functions. Antioxidants, such as ascorbic acid (AA), are free radical scavengers and improve the immune response. In the pathogenesis of endotoxic shock, a disease with high mortality caused by gram-negative bacterial endotoxin, the reactive oxygen species (ROS) produced by phagocytes have been implicated. In a previous study, we observed in peritoneal macrophages from BALB/c mice suffering lethal endotoxic shock caused by intraperitoneal (i.p.) injection of Escherichia coli lipopolysaccharide (LPS; 100 mg/kg) a high production of superoxide anion. Therefore, in the present work, we have studied the in vitro effect of AA, at different concentrations (0.001, 0.01, 0.1, 1 and 2.5 mM), on the various steps of the phagocytic process, i.e., adherence to substrate, chemotaxis, ingestion of particles and superoxide anion production of murine peritoneal macrophages obtained from BALB/c mice with that of endotoxic shock, at 2, 4, 12 and 24 h after LPS injection. The increased adherence, ingestion and superoxide anion production by macrophages from animals with endotoxic shock were lower in the presence of AA, reaching similar values to those of the control animals. The most effective AA concentration in cells from mice with endotoxic shock was 0.01 mM. These data suggest that AA can regulate the phagocytic process in endotoxic shock, principally decreasing free radical production and thus it could reduce endotoxic shock severity. PMID- 10665784 TI - Carcinoma of the cervix and the use of hyperbaric oxygen with radiotherapy: a report of a randomised controlled trial. AB - BACKGROUND AND PURPOSE: A randomised controlled trial of hyperbaric oxygen in the radiotherapy of Stage IIb and III carcinoma of cervix was performed between 1971 and 1980. Apart from an abstract giving an interim report in 1977, results have not been published. MATERIAL AND METHODS: In a four arm study, 335 patients were randomised to treatment in 10 or 28 fractions, in hyperbaric oxygen or in air. Data is available concerning 327 cases and this has been analysed. RESULTS: There was no advantage in tumour control shown with the use of hyperbaric oxygen. There was evidence for an increase in late radiation morbidity when treatment was given in hyperbaric oxygen rather than in air and when, using 10 fractions, a total dose of 45 rather than 40 Gy was achieved. For late intestinal morbidity, the fractionation sensitivity (alpha/beta ratio) was calculated to be 4.3 Gy and the steepness of the dose response curve (gamma50) to be 2.6. CONCLUSIONS: Hyperbaric oxygen gave no benefit in the treatment of patients with stage IIb and III carcinoma of the cervix treated with radiotherapy using two fractionation regimes. Important data regarding late radiation morbidity has been revealed. PMID- 10665785 TI - Intratumoral pO2-measurements as predictive assay in the treatment of carcinoma of the uterine cervix. AB - BACKGROUND: Several studies have shown that pretreatment oxygenation status of cervical tumors measured with a polarographic oxygen electrode could be a predictive factor for radiation response and survival. The purpose of this study was to evaluate the impact of intratumoral pO2 levels and hypoxic fractions on local control and disease free survival employing a standardized measuring procedure under routine conditions. MATERIALS AND METHODS: Between April 1994 and December 1997 pO2 measurements were performed prior to radiotherapy with an Eppendorf histograph in 51 evaluable patients with primary cervical carcinoma. All patients were treated with curative intent by combined external beam therapy (median total dose 49.6 Gy) and 3-6 applications of high dose rate- (7 Gy/fr. at point 'A') or pulse dose rate brachytherapy (20-25 h pulses, 1 Gy/pulse at point 'A'). Oxygenation data are given as median pO2 of pooled readings and percentage of readings below 5 mm Hg (HF 5). RESULTS: Median pO2 values ranged from 0 to 60 mm Hg (median 10). HF5 ranged from 0 to 95% (median 22%). Median follow-up was 26 months (range 9-54 months). Actuarial overall and disease-free survival rates (OS/DFS) at 3 years were 53%/50%. Comparing patients with median pO2 < or = 10 mm Hg (n = 26) to patients with higher median pO2 levels (n = 25) calculated DFS was 34 and 69%, respectively (P < 0.02). Corresponding data for local control were 47 and 84% (P = 0.053). Comparing patients with HF5 below and above the median calculated DFS was 36 and 66%, respectively (P < 0.02). Patients who had median pO2 < 10 mm Hg and HF5 > 20% had the worst prognosis (3-year DFS: 28%). Besides oxygenation status, stage and initial hemoglobin concentration were statistically significant for treatment outcome. CONCLUSIONS: This study confirms earlier data that the presence of hypoxia is associated with poor local control and survival in patients with carcinoma of the uterine cervix. Polarographic pO2 measurements are feasible under routine conditions and can be regarded as a reproducible predictive assay. PMID- 10665786 TI - Tumoural perfusion as measured by dynamic computed tomography in head and neck carcinoma. AB - PURPOSE: To investigate the intra- and interobserver variability, as well as the intra- and interpatient variability of CT-determined tumour perfusion in head and neck tumours, and to evaluate the preliminary value of this parameter as predictive factor of local failure after treatment by definitive radiotherapy. MATERIALS AND METHODS: In 41 patients the perfusion of a primary head and neck squamous cell carcinoma was estimated using dynamic CT. A 40-ml intravenous bolus of a low-osmolar non-ionic contrast agent was rapidly injected over 5 s (8 ml/s), while a dynamic acquisition of image data was obtained during the first pass at the level of the largest axial tumour surface. A time-density curve was constructed for the primary tumour and the carotid artery. The perfusion in the selected tumour region of interest was calculated by dividing the slope of the tumour-time density curve by the maximal value in arterial density. Tumour volume was calculated on the CT-images and correlated with perfusion rate. RESULTS: The mean perfusion rate was 86.4 ml/min per 100 g (median, 80.6; SD, 43.05; range, 31.7-239.8 ml/min per 100 g). No systematic difference was found between the measurements performed by two independent observers. The intratumoural COV was 0.22, the intertumoural COV 0.37. No correlation was found with tumour volume. Ten out of 20 patients with a perfusion rate < 80 ml/min per 100 g were not locally controlled, while nine out of 21 patients with a value > 80 ml/min per 100 g did show a local failure (P = 0.19). CONCLUSIONS: CT-determined perfusion measurements of head and neck tumours are feasible. No correlation with tumour volume and a sufficiently large COV were found to consider this parameter as a possible prognostic factor for outcome after radiotherapy. More patients need to be investigated to test the hypothesis that tumours with a low CT determined perfusion rate have a higher risk of local failure. PMID- 10665787 TI - Oxygenation of head and neck cancer: changes during radiotherapy and impact on treatment outcome. AB - BACKGROUND AND PURPOSE: To evaluate the long term clinical significance of tumor oxygenation in a population of head and neck cancer patients receiving radiotherapy and to assess changes in tumor oxygenation during the course of treatment. METHODS AND MATERIALS: Patients with head and neck cancer receiving primary RT underwent pretreatment polarographic tumor oxygen measurement of the primary site or a metastatic neck lymph node. Treatment consisted of once daily (2 Gy/fraction to a total dose of 66-70 Gy) or twice daily irradiation (1.25 Gy/fraction to 70-75 Gy) to the primary site. Twenty-seven patients underwent a second series of measurements early in the course of irradiation. RESULTS: Sixty three patients underwent pretreatment tumor oxygen assessment (primary site, n = 24; nodes, n = 39). The median pO2 for primary lesions was 4.8 mmHg, and it was 4.3 mmHg for cervical nodes. There was a weak association between anemia and more poorly oxygened tumors, but many non-anemic patients still had poorly oxygenated tumors. Repeat assessments of tumor oxygenation after 10-15 Gy were unchanged compared to pretreatment baselines. Poorly oxygenated nodes pretreatment were more likely to contain viable residual disease at post-radiation neck dissection. Median follow-up time for surviving patients was 20 months (range 3-50 months). Hypoxia (tumor median pO2 <10 mmHg) adversely affected 2 year local-regional control (30 vs. 73%, P = 0.01), disease-free survival (26 vs. 73%, P = 0.005), and survival (35 vs. 83%, P = 0.02). CONCLUSION: Tumor oxygenation affects the prognosis of head and neck cancer independently of other known prognostic variables. This parameter may be a useful tool for the selection of patients for investigational treatment strategies. PMID- 10665788 TI - Prognostic impact of total tumor volume and hemoglobin concentration on the outcome of patients with advanced head and neck cancer after concomitant boost radiochemotherapy. AB - PURPOSE: To identify prognostic clinical and treatment related factors for local control, distant metastasis-free survival, and survival by means of a multivariate analysis in patients with advanced squamous cell carcinoma of the head and neck after concomitant boost radiochemotherapy. PATIENTS AND METHODS: From 1992 to 1995, 68 patients with squamous cell cancer of the head and neck (93% stage IV disease) were treated with a simultaneous radiochemotherapy with Carboplatin using a concomitant boost technique. The total tumor volume (TTV) was quantitatively determined based on computed tomography scans in 56 patients. A Cox proportional hazards regression analysis was performed for each of the above endpoints and statistical significance of the Cox models was verified using the likelihood ratio test and Bonferroni correction for multiple testing. RESULTS: The survival and locoregional control rates at three years were 35 and 32%. The multivariate analysis revealed a significant association between the TTV and survival (P = 0.0008) and between the pretreatment serum hemoglobin concentration and locoregional control (P = 0.01) and survival (P = 0.05). The locoregional control was significantly associated with the N-stage (P = 0.007) and there was a good correlation between the N-stage and TTV in this study population. CONCLUSION: Our data corroborate the prognostic relevance of the tumor volume and hemoglobin concentration. In studies comparing the survival of patients with advanced cancer of the head and neck, the use of the TTV as a covariable may improve the statistical power. PMID- 10665789 TI - Polarographic measurements of oxygen tension in human glioma and surrounding peritumoural brain tissue. AB - This study quantifies the spatial distribution of pO2 in glioma and in the surrounding brain tissue. Both glioma and peritumoural brain contain regions at oxygen tensions less than 2.5 mmHg. Modalities targeting hypoxia to improve the efficacy of therapy may have an important role in the management of this disease. PMID- 10665790 TI - Results of ruthenium irradiation of uveal melanomas: the Dutch experience. AB - BACKGROUND AND PURPOSE: To update our results of the treatment of uveal melanomas using ruthenium applicators in 49 patients treated with graded doses and subsequently in 52 patients with maximal scleral doses of 800 Gy and an effective top dose of at least 160 Gy. MATERIAL AND METHODS: One hundred and one patients were treated with brachytherapy only, in 25 patients it was combined with transpupillary thermotherapy (TTT). RESULTS: A complete remission was found in 62.6% of the patients and in 31.3% a stable disease with an average follow-up of 74.6 months. Above a top dose of 120 Gy only in one of 95 patients continuous tumour growth after treatment was observed. Useful vision could be preserved in 51.5% of the patients. The initial tumour prominence and top dose strongly correlated with treatment outcome. CONCLUSIONS: Ruthenium application for uveal melanomas with the doses we have used is successful, with a substantial number of patients having their eyes preserved, their tumour controlled and their vision retained. Further improvements can be obtained with ruthenium irradiation with lower maximal scleral doses combined with TTT. PMID- 10665791 TI - Radiotherapy of unilateral choroidal metastasis: unilateral irradiation or bilateral irradiation for sterilization of suspected contralateral disease? AB - Radiotherapy is the highly effective standard in the treatment of choroidal metastasis. Visual acuity can be stabilized or increased in about 70-80% of eyes treated, thus prevailing the quality of life in these worse prognostic patients. In about 30-40% bilateral macroscopic disease is found at diagnosis. The best treatment for unilateral metastasis remains controversial: unilateral or bilateral irradiation for sterilization of suspected contralateral metastasis or unilateral irradiation without irradiation of the contralateral choroidea. In the analysis of a prospective study (ARO 95-08) 35 out of 50 patients with choroidal metastasis had unilateral disease and received unilateral irradiation with a lateral field using 6 MeV-photons (40 Gy in 20 fractions) without sparing the contralateral choroidea. Therefore the posterior contralateral choroidea received 50-70% of the total dose (20-28 Gy) for suspected micrometastasis. None of these patients developed contralateral choroidal metastasis during the median follow up time of 11.5 months. A unilateral field with 40 Gy for unilateral choroidal metastasis without sparing the contralateral choroidea seems to be effective in destroying contralateral micrometastasis with a lower risk of late side effects compared with bilateral fields. PMID- 10665792 TI - Radiation late effects in children treated for orbital rhabdomyosarcoma. AB - BACKGROUND AND PURPOSE: The experience resulting from large cooperative studies shows that correct radiation therapy at doses adequate to the tumor bulk are crucial for local control of rhabdomyosarcoma. The aim of the present study was to document the correlation between modalities and doses of radiotherapy and radiation side effects. PATIENTS AND METHODS: Between 1980 and 1997, 19 patients affected by primary orbital rhabdomyosarcoma have been followed at the University Federico II of Naples. All but three patients, who received 45, 54 and 55 Gy respectively, have been treated by immediate radiation at the dose of 60 Gy, delivered in 2 Gy fractions, five times per week, by cobalt 60 megavoltage equipment. Combined chemotherapy using vincristine and vincristine plus dactinomycin on alternate weeks was also administered as part of induction therapy. RESULTS: An overall survival rate of 94.7% was registered. In our patients the majority of radiation late effects were paid by orbit and ocular adnexa. Side effects to lens and ocular structures were fewer and of low grade. CONCLUSIONS: Radiation therapy is still essential for local control of orbital rhabdomyosarcoma, however radiation side effects have to be carefully considered together with the therapeutic goal to be obtained. PMID- 10665793 TI - Evaluation of frequency and type of errors detected by a computerized record and verify system during radiation treatment. AB - BACKGROUND: Computerized record and verify systems (RVS) have been introduced to improve the precision of radiation treatment delivery. These systems prevent the delivery of ionizing radiations when the settings of the treatment machine do not match the intended parameters within some maximal authorized deviation. PURPOSE: To assess the potential alteration of the frequency of errors associated with the use of RVS during radiation treatment delivery. MATERIALS AND METHODS: The software of the RVS was altered in order to record the settings actually used for radiation treatment delivery whereas the verification function was suppressed. At the end of the study period, the settings used during daily administration of radiation treatment were compared to the parameters recorded in the RVS using the computer. They were also compared with the planned ones written in the patient treatment chart. RESULTS: Out of the 147,476 parameters examined during the study period, 678 (0.46%) were set erroneously. At least one error occurred in 628 (3.22%) of the 19,512 treated fields. An erroneous parameter was introduced in the RVS memory in 22 (1.17%) of the 1885 fields. CONCLUSIONS: RVS has the potential to improve precision of radiation treatment delivery by detecting a significant number of setting errors. However, excessive confidence in RVS could lead to repeated errors as there is a potential for the entry of erroneous parameters into the RVS memory. PMID- 10665794 TI - Diamond detector measurements near simulated air channels for narrow photon beams. AB - BACKGROUND AND PURPOSE: To evaluate the combined effect of increased photon transmission, reduced photon scatter, increased secondary electron range and loss of electronic equilibrium for narrow 6-MV beams in and around a simulated air channel. MATERIALS AND METHODS: A measuring method was developed in-house for relative dose measurements near simulated air-like/soft-tissue interfaces in an automated water phantom. A Styrofoam cylinder (density 0.03 g/cm3) of 2-cm diameter was submersed in the water phantom and irradiated with small rectangular radiation fields. The field length was fixed at 10 cm and the field widths ranged from 1 to 4 cm. The axis of the foam cylinder and the long side of the field were parallel. A water layer of 2 cm was realised upstream of the cylinder. Relative depth dose and profiles behind the foam cavity were assessed using a diamond detector with a sensitive crystal thickness of 0.21 mm located at 1 mm from the top of the encapsulation. RESULTS: The dose at central axis 1.1 mm behind the cavity was found to be 92 and 74% for a field size of 10 x 2 and 10 x 1 cm2, respectively. The highly convex dose profile of the 10 x 1-cm2 field, characterising the homogeneous case, is flattened. CONCLUSIONS: The diamond detector is an excellent choice as a detector in small photon fields with high dose gradients as they occur near air channels, provided the orientation of the detector is appropriate. Doses near air channels are subject to significant local variations as a function of small changes of field width, and local underdosing may occur in particular cases. PMID- 10665795 TI - The use of high density metal foils to increase surface dose in low-energy clinical electron beams. AB - BACKGROUND AND PURPOSE: This paper describes a practical method of elevating the surface dose of clinical electron beams in the energy range 3-12 MeV using thin high density metal foils (tin and lead) as an alternative to tissue equivalent bolus. Because, relative to water, these materials exhibit a high scattering power to stopping power ratio, the desired dose elevation may be achieved with less energy loss than conventional bolus and consequently a gain in therapeutic interval. METHODS: The foil thickness required to raise the surface dose to 90% off peak, for a given electron energy, was calculated using published scattering and stopping power data. An empirical expression is derived to facilitate calculation of foil thickness (tin or lead) to produce a given surface dose. RESULTS AND CONCLUSIONS: Measurements were made to confirm the predictions of the derived expression and were found to be in good agreement. PMID- 10665796 TI - Is tourniquet use ineffective in the pre-hospital management of South American rattlesnake bite? PMID- 10665797 TI - Cloning and functional expression of a synthetic gene encoding huwentoxin-I, a neurotoxin from the Chinese bird spider (Selenocosmia huwena). AB - Cloning and functional expression of a synthetic gene encoding huwentoxin-I, a neurotoxin from the Chinese bird spider Selenocosmia huwena. A gene encoding huwentoxin-I, a peptide neurotoxin consisted of 33 amino acid residues from the venom of the Chinese bird spider Selenocosmia huwena, was designed, synthesized and expressed in Escherichia coli as a hybrid protein fused with glutathione S transferase at the N-terminal. The fusion protein was purified by GSH-Sepharose 4B affinity column chromatography and cleaved by thrombin to release the toxin peptide. The amino acid sequence of the recombinant toxin was consistent with the designed one by sequence determination and MALDI-TOF mass analysis, suggesting that the recombinant huwentoxin-I produced the same expression product as the native one. After reduction and renaturation, the biological activity of the recombinant toxin was identical with that of the native huwentoxin-I by electrophysiological method. PMID- 10665798 TI - Some biochemical properties of Russell's viper (Daboia russelli) venom from Eastern India: correlation with clinico-pathological manifestation in Russell's viper bite. AB - In the present study, some biochemical properties and pathological effects of Daboia russelli venom from Burdwan district of West Bengal, eastern India are presented. The clinical features of Russell's viper envenomation observed in patients admitted to Burdwan Medical College & Hospital are also reported. In vitro, whole venom exerts strong trypsin inhibitory, phospholipase A2 and procoagulant activities in addition to moderate adenosine monophosphatase and adenosine triphosphatase activities. Lethality (LD50) of this venom sample is 0.7 mg kg (i.v.) of mice. Significant local tissue damaging effects including edema, hemorrhage and necrosis are observed in experimental animal models. An increase in the level of serum enzymes, such as aspartate transaminase, alkaline phosphatase, creatine phosphokinase, lactate dehydrogenase after D. russelli venom injection in albino rats is indicative of cell or tissue damage. High incidence of intravascular hemolysis in addition to hemostasis, haemoptysis and haematuria are observed as the most prominent features of RVV envenomation from this part of India. The present study reinforces the hypothesis that variation in the venom composition of RVV from eastern India with respect to venom samples of Russell's vipers from other parts of India is responsible for the differences in the clinical manifestation in patients from eastern India. PMID- 10665799 TI - Beta-agkistrodotoxin inhibits fast and Ca2+-activated K+ currents recorded from mouse motor nerve terminals. AB - Beta-agkistrodotoxin (beta-AgTx), a polypeptide purified from the venom of Agkistrodon blomhoffii brevicaudus, is a presynaptic blocker acting on neurotransmitter release. In this work, perineural recording technique was employed to study the effects of beta-AgTx on sodium, potassium and calcium currents of mouse motor nerve terminals. The results showed that beta-AgTx selectively inhibited Ca2+-dependent (I(K,Ca)) and fast (I(K,f) K+ currents, but did not affect slow K+ current (I(K,s)), sodium and calcium currents. However there are other components in A. blomhoffii brevicaudus venom which inhibit perineural sodium current. The present data have provided additional evidence that the site of action of beta-AgTx is different from that of beta-bungarotoxin. PMID- 10665800 TI - m1-toxin isotoxins from the green mamba (Dendroaspis angusticeps) that selectively block m1 muscarinic receptors. AB - The venom of the green mamba, Dendroaspis angusticeps, was found to have at least six trace m1-specific toxins that block the binding of 3H-N-methylscopolamine to cloned m1 muscarinic receptors. Four were isolated by gel filtration, cation exchange HPLC and reversed-phase HPLC and named m1-toxins1-4. Recovery was 180, 90, 20 and 10 microg/g dry venom, respectively. m1-Toxin1 (the original m1-toxin) was found to have the sequence, L T C V K S N S I W F P T S E D C P D G Q N LC F K R W Q Y I S P R M Y D F T R G C A A T C P K A E Y R D V I N C C G T D K C N K, calculated mass = 7473 Da and calculated pI = 8.2. This sequence had been predicted previously from a cDNA cloned from the venom glands of this snake. The binding of m1-toxin1 was irreversible, so its Kd could not be determined. m1 Toxin2 differed only in proline-19, mass = 7455 and pI = 8.5. Partial sequence data for m1-toxin3 showed aspartate-7 and m1-toxin4 showed isoleucine-12, asparagine-16 and alanine-19. m1-Toxins1-4 have seven conserved amino acids not found in homologous mamba toxins that bind to other muscarinic receptors (MT1, MT2, m4-toxin = MT3, MT4, MT5, MTalpha and MTbeta). Some of these residues may be essential for m1-specificity. Since m1-toxin1 binds irreversibly in artificial cerebrospinal fluid at 37 degrees C, it is a particularly attractive antagonist for studies in vivo. PMID- 10665801 TI - Effect of crotapotin and heparin on the rat paw oedema induced by different secretory phospholipases A2. AB - The effects of crotapotin (a non-toxic and non-enzymatic acid polypeptide naturally complexed with phospholipase A2) and heparin on rat paw edema induced by different secretory phospholipases A2 (sPLA2) have been investigated. The ability of crotapotin to affect the enzymatic activity of the sPLA2(s) have also been evaluated. Secretory PLA2(s) obtained from both snake (Naja naja, Naja mocambique mocambique, Crotalus adamanteus and Crotalus durissus terrificus) and bee (Apis mellifera) venoms as well as that from bovine pancreas were used in this study. Rat paw oedema was induced by a single subplantar injection of the sPLA2s (5-30 microg/paw) in absence and presence of either crotapotin (10-100 microg/paw) or heparin (50 U/paw). Paw volume was measured using a hydroplethysmometer. Phospholipase A2 from Naja naja, Naja mocambique mocambique, Apis mellifera venoms and the basic component of Crotalus durissus terrificus venom all induced dose-dependent rat paw oedema whereas those from Crotalus adamanteus venom and bovine pancreas were ineffective. Paw oedema induced by PLA2(s) from both Naja naja and Apis mellifera venoms was significantly (P < 0.05) inhibited by crotapotin (0.1-100 microg/site) whereas the Naja mocambique mocambique venom PLA2-induced oedema was significantly potentiated (P < 0.05) by this polypeptide (40 microg/site). On the other hand, heparin (50 U/paw) had no effect on the paw oedema induced by PLA2 from Naja naja and Apis mellifera venoms but significantly inhibited the Naja mocambique mocambique venom PLA2-induced oedema. The measurement of the in vitro phospholipasic activity revealed that crotapotin inhibited by 60-70% the enzymatic activities of PLA2(s) from Crotalus adamanteus, Naja mocambique mocambique, Apis mellifera venoms and bovine pancreas. Our results suggest that despite the great homology between the various types of sPLA2 they interact with crotapotin on cell surfaces in different ways leading to either inhibition or potentiation of the paw oedema by a mechanism unrelated to their enzymatic activities. Since heparin reduced paw oedema induced by PLA2 from Naja mocambique mocambique venom it is likely that this sPLA2 is similar to the novel heparin-sensitive PLA2 found in mast cells. PMID- 10665802 TI - Bothrops lanceolatus (Fer de lance) venom induces oedema formation and increases vascular permeability in the mouse hind paw. AB - The ability of snake venoms to increase vascular permeability and to induce oedema through the release of pharmacologically active substances is well known. We have studied the oedema and vascular permeability induced by Bothrops lanceolatus venom in male Swiss white mice. Paw oedema was induced by the subplantar injection of B. lanceolatus venom (125-1000 ng/paw) and was quantified as the increase in paw weight. Changes in vascular permeability were assessed by measuring the amount of Evans blue dye extravasation. The oedema and the increase in vascular permeability were maximal within 2 h and had resolved after 24 h. The administration of the vasodilator iloprost (20 ng/paw) immediately after B. lanceolatus venom potentiated the oedema and the increase in vascular permeability by approximately four-fold. Pretreating the mice with indomethacin, dexamethasone, NDGA or BW A4C inhibited the venom-induced oedema and the increase in vascular permeability. In contrast, histamine, serotonin and PAF-acether antagonists (mepyramine, cyproheptadine and WEB 2086, respectively) were ineffective. Histological examination showed that B. lanceolatus venom (250 ng and 500 ng/paw) caused thickening of the inner dermal layers which was accompanied by extensive intercellular spaces indicative of oedema. In addition, there was a marked infiltration of inflammatory cells, particularly neutrophils, into the underlying muscle layer. The latter, however, remained morphologically unaffected during the 3 h of observation. Venom doses larger than 500 ng/paw produced intense haemorrhage. These results indicate that B. lanceolatus venom induces oedema and increases vascular permeability in the mouse hind paw. The principal mediators of this inflammatory response are cyclooxygenase and lipoxygenase products. PMID- 10665803 TI - Venom alkaloids of fire ants in relation to worker size and age. AB - Piperidine alkaloids compose most of the venom of the red imported fire ant, Solenopsis invicta, and we examined how six of these alkaloids varied across worker size and age. In a colony sampled intensively, the relative abundance of each alkaloid was highly correlated with worker size with one exception, and ratios of saturated to unsaturated alkaloids were positively correlated with worker size. Similarly, both the abundance and ratios of alkaloids differed significantly between the small and large workers sampled from colonies across Texas, USA. Young and old workers produced less venom than ants of intermediate age (3-7 weeks), and ratios of saturated to unsaturated alkaloids increased significantly with worker age. The differences in venom composition correspond to the size- and age-based functional roles of workers. PMID- 10665804 TI - Comparative study on the ability of IgG and Fab sheep antivenoms to neutralize local hemorrhage, edema and myonecrosis induced by Bothrops asper (terciopelo) snake venom. AB - The ability of sheep antivenoms, consisting of whole IgG molecules or Fab fragments, to neutralize local hemorrhage, edema and myonecrosis induced by Bothrops asper venom was comparatively studied in mice. The two antivenoms were produced from the same batch of hyperimmune plasma and were adjusted to the same neutralizing potency against these effects in assays where venom and antivenoms were incubated prior to injection. Thus, if differences are observed in experiments involving independent injection of venom and antivenoms, they would depend on the pharmacokinetic profiles of the products. Despite the observation that both antivenoms neutralized the three effects if preincubated with venom, neutralization was only partial when antivenoms were administered i.v. at various time intervals after envenomation. No significant differences were observed between IgG and Fab antivenoms concerning neutralization of hemorrhagic and edema forming activities, whereas IgG antivenom was slightly more effective in neutralizing myotoxic activity in experiments involving independent injection of venom and antivenom. These results do not support the hypothesis that Fab fragments are more effective than whole IgG molecules in the neutralization of locally-acting toxins from B. asper venom. PMID- 10665805 TI - Sensitivity of embryonic rat dorsal root ganglia neurons to Clostridium botulinum neurotoxins. AB - Clostridium botulinum neurotoxins (BoNT) are zinc dependent endopeptidases which, once internalised into the neuronal cytosol, block neurotransmission by proteolysis of membrane-associated proteins putatively involved in synaptic vesicle docking and fusion with the plasma membrane. Although many studies have used a variety of cellular systems to study the neurotoxins, most require relatively large amounts of toxin or permeabilisation to internalise the neurotoxin. We present here a primary culture of embryonic rat dorsal root ganglia (DRG) neurons that exhibits calcium-dependent substance P secretion when depolarised with elevated extracellular potassium and is naturally BoNT sensitive. The DRG neurons showed a different IC50 for each of the toxins tested with a 1000 fold difference between the most and least potent neurotoxins (0.05, 0.3, 30 and approximately 60 nM for A, C, F and B, respectively). BoNT/A cleavage of SNAP-25 was seen as early as 2 h, but substance P secretion was not significantly inhibited until 4 h intoxication and the effects of BoNT/A were observed for as long as 15 days. This primary neuronal culture system represents a new and sensitive cellular model for the in vitro study of the botulinum neurotoxins. PMID- 10665806 TI - Amino acid sequence studies on cytolytic toxins from sea anemone Heteractis magnifica, Entacmaea quadricolor and Stichodactyla mertensii (Anthozoa). AB - The complete amino acid sequence of a cytolytic toxin, HmT, isolated from sea anemone Heteractis magnifica was determined. It is composed of 177 amino acid residues and lacks half-cystines. Partial N-terminal sequences of three other cytolysins from Entacmaea quadricolor (EnT) and Stichodactyla mertensii (SmT-1 and SmT-2) were also determined. Comparing these sequences with those of other sea anemone cytolysins, a high degree of homology was observed. PMID- 10665807 TI - Structural and ultrastructural description of the venom gland of Loxosceles intermedia (brown spider). AB - The brown spider, genus Loxosceles, is becoming of great medical importance, with envenomation (Loxoscelism) occurring throughout the world. The biological activities of the brown spider venom usually include dermonecrotic lesions at the bite site accompanied by hemolytic and haemorrhagic effects and also by renal failure. The objective of the present study was to describe the histology of the venom gland of L. intermedia using glands from adult spiders which were investigated by light microscopy, using immunohistochemical and staining methods, by transmission electron microscopy, and by scanning electron microscopy. The organization of the venom gland of Loxosceles intermedia follows the general architecture of spiders' venom glands. Using light microscopy and transmission electron microscopy we observed that the venom glands of L. intermedia present two layers of striated muscle fibers, an external layer and an internal layer in touch with an extracellular matrix which is a basement membrane structure and a fibrillar collagen matrix separating the muscular region from epithelial cells of the venom gland. Muscle cells are multinucleated, with nuclei peripherally placed and their cytoplasm rich in sarcoplasmic reticulum, myofibrills and continuous Z lines. By using scanning electron microscopy we can detect muscular cells from external layer as branching cells. Epithelial cells have their cytosol extremely rich in rough endoplasmic reticulum, mitochondria collection, Golgi apparatus, interdigitating membranes and secretory vesicles that ultimately accumulate the venom, a complex protein mixture. PMID- 10665808 TI - An investigation of the degradation of the plant toxin, ricin, by sodium hypochlorite. AB - The toxin, ricin (0.4 microg/microl), was exposed to a range of sodium hypochlorite concentrations. SDS PAGE showed that hypochlorite caused the ricin to smear and decrease in mobility and, ultimately, caused a loss of silver staining. Cytotoxicity assays using dye uptake by Hep2 cells showed that treatment with 3 mM hypochlorite inactivated the ricin. Western blotting and ELISAs showed that binding by polyclonal antibodies raised against native ricin, or partially degraded ricin, diminished as hypochlorite degradation of the ricin increased. PMID- 10665809 TI - The influence of hunger and breeding temperature on the venom production of the spider Cupiennius salei (Araneae, Ctenidae). AB - The venom production of the free hunting neotropic spider Cupiennius salei was tested under different breeding conditions. Three groups kept at different temperatures (17, 21 and 25 degrees C) showed that venom production remained stable within this temperature range, only at a temperature of 15 degrees C the spiders stopped feeding and venom synthesis. Hunger periods do not have a direct effect on the released venom quantity. Two groups of spiders--the first group after a four and the second after an eight weeks hunger period--were compared and no difference in venom production was found. Such long fasting periods are a natural situation for spiders. In this case Cupiennius salei reduces its body weight but not venom supply. This means that body weight is a parameter only of short-term fitness which changes with the actual living conditions (temperature, feeding intervals) of each individual. Long-term fitness is best described by the prosoma length, which was formed during the juvenile growth of each spider and is rather invariable in adulthood. It was shown that the quantity of released venom is better correlated with the length of the prosoma than with the weight of the animal. This means that venom production is mostly an indicator of long-term fitness. PMID- 10665810 TI - Notes on the traditional use of plants to treat snake bite in northern Papua New Guinea. AB - Information on snake bite treatment using plants was collected in three areas of northern Papua New Guinea. Liquid from six plants is either placed topically on the bite site or plant parts (bark) are chewed and the sap is swallowed. However, beside tannins in the bark no secondary metabolites, e.g. alkaloids, steroids, saponins or flavonoids have been detected by thin-layer chromatography in alcoholic extracts of the plants involved rendering this type of snake bite treatment questionable. PMID- 10665811 TI - The adsorption of microcystin-LR by natural clay particles. AB - The microcystin cyanobacterial hepatotoxins represent an increasingly severe global health hazard. Since microcystins are found world wide in drinking water reservoirs concern about the impact on human health has prompted investigations into remedial water treatment methods. This preliminary study investigates the scavenging from water of microcystin-LR by fine-grained particles known to have a high concentration of the clay minerals kaolinite and montmorillonite. The results show that more than 81% of microcystin-LR can be removed from water by clay material. Thus, microcystin-LR is indeed scavenged from water bodies by fine grained particles and that this property may offer an effective method of stripping these toxins from drinking water supplies. PMID- 10665812 TI - Hepatotoxicity of Eupatorium adenophorum to rats. AB - Freeze dried Eupatorium adenophorum leaf powder mixed in rat feed at a level of 25% elicited hepatotoxicity. The affected animals were jaundiced and had marked increase in plasma bilirubin levels and activities of alkaline phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase. The liver of intoxicated animals had focal areas of necrosis and bile duct proliferation. Elevation in plasma bilirubin concomitant with alterations in enzyme profile and histopathological lesions are consistent with liver injury and cholestasis. This is the first report of the toxicity of E. adenophorum to rats. PMID- 10665813 TI - Bibliography of toxinology. PMID- 10665814 TI - Determinants of voltage-dependent inactivation affect Mibefradil block of calcium channels. AB - The voltage gated calcium channel family is a major target for a range of therapeutic drugs. Mibefradil (Ro 40-5967) belongs to a new chemical class of these molecules which differs from other Ca2+ antagonists by its ability to potently block T-type Ca2+ channels. However, this molecule has also been shown to inhibit other Ca2+ channel subtypes. To further analyze the mechanism governing the Ca2+ channel-Mibefradil interaction, we examined the effect of Mibefradil on various recombinant Ca2+ channels expressed in mammalian cells from their cloned cDNAs, using Ca2+ as the permeant ion at physiological concentration. Expression of alpha1A, alpha1C, and alpha1E in tsA 201 cells resulted in Ca2+ currents with functional characteristics closely related to those of their native counterparts. Mibefradil blocked alpha1A and alpha1E with a Kd comparable to that reported for T-type channels, but had a lower affinity (approximately 30-fold) for alpha1C. For each channel, inhibition by Mibefradil was consistent with high-affinity binding to the inactivated state. Modulation of the voltage-dependent inactivation properties by the nature of the coexpressed beta subunit or the alpha1 splice variant altered block at the Mibefradil receptor site. Therefore, we conclude that the tissue and sub-cellular localization of calcium channel subunits as well as their specific associations are essential parameters to understand the in vivo effects of Mibefradil. PMID- 10665815 TI - A point mutation in the maxi-K clone dSlo forms a high affinity site for charybdotoxin. AB - This work investigated the interaction of CTX with two cloned analogues of the maxi-K channel, dSlo and hSlo. dSlo has been reported to be CTX insensitive. Single channel analysis revealed that dSlo was weakly blocked by the toxin, with a very high K(D) of 5.8 microM. The hSlo channels bound wild-type, recombinant CTX with high affinity and in a bimolecular fashion, and displayed a half blocking concentration (K(D)) of 36 nM. A glutamate residue was substituted for the wild-type threonine at position 290 in dSlo. The mutant channel was expressed in COS-7 cells and reconstituted into lipid bilayers for single channel analysis. The mutant channel bound wild-type, recombinant CTX with high affinity and in a bimolecular fashion, and displayed a half-blocking concentration (K(D)) of 23 nM. Changing just one residue from threonine to glutamate at position 290 in dSlo changed the affinity of the channel's CTX-receptor over 100-fold. Kinetic analysis revealed that this large increase in affinity was due to decreasing the dissociation rate of the toxin. These results suggest that a CTX receptor does exist in the dSlo channel mouth and that the threonine at position 290 destabilizes the toxin on the binding site. PMID- 10665816 TI - The contributions of GluR2 to allosteric modulation of AMPA receptors. AB - Native AMPA receptor complexes in the CNS are composed of hetero-oligomers of the GluR1-4 subunits, and generally contain the GluR2 subunit. To determine the contributions of GluR2 to pharmacological properties of receptor complexes, the effect of hetero-oligomerization with GluR2 on allosteric modulation of recombinant AMPA receptors was studied. The study of homo-oligomeric GluR2 was facilitated with a site-directed mutant of the pore, GluR2(R607Q), which allowed robust currents from this normally low-conducting subunit. The efficacy of the allosteric modulators was tested on homo-oligomeric GluR1-4, and then compared with hetero-oligomeric GluR1/GluR2, GluR3/GluR2 and GluR4/GluR2. Two selective allosteric modulators were tested, a positive modulator, cyclothiazide, and a negative modulator, LY300164. The results show that the pharmacological properties of homo-oligomeric GluR2 are not significantly different from those of GluR1, GluR3 or GluR4. The apparent affinity of cyclothiazide is not significantly changed upon hetero-oligomerization. However, the extent of potentiation of kainate responses by cyclothiazide is significantly decreased upon hetero-oligomerization. Hetero-oligomerization increases the apparent affinity of LY300164, a (-) isomer of the 2,3-benzodiazepine LY293606. These data indicate that although GluR2 has a dominant effect on the permeation properties, this subunit does not have a similarly dominant effect on pharmacological properties of native receptors. However, the state of hetero-oligomerization can alter the pharmacological properties of AMPA receptors. PMID- 10665817 TI - Cocaine-induced kindling is associated with elevated NMDA receptor binding in discrete mouse brain regions. AB - The present study was undertaken to investigate the involvement of N-methyl-D aspartate (NMDA) type of glutamate receptors in the induction and maintenance of kindling generated by daily cocaine (35 mg/kg) injections to Swiss Webster mice. In addition, the regulation of NMDA receptor binding following the development of sensitization to horizontal locomotor activity produced by daily injections of a low dose of cocaine (15 mg/kg for 5 days) was investigated. Three days following the administration of the high dose of cocaine (35 mg/kg) a marked augmentation in cocaine-induced horizontal and vertical activities was observed (induction phase). Subsequently, after 10 days of cocaine administration, mice developed stage 5 seizures (Racine scale). Binding of [3H]CGP 39653 to the NMDA receptors revealed a marked increase in receptor densities in the striatum, amygdala and hippocampus associated with the induction phase. The elevation of NMDA receptor binding in the striatum and amygdala was sustained for 10 days following the induction phase. The pattern of altered NMDA receptor binding following the expression of cocaine kindled seizures was different. One day after the expression of kindled seizures NMDA receptor binding was elevated in striatum, amygdala, hippocampus and frontal cortex. However, only the elevation of NMDA receptor binding in the amygdala and hippocampus was sustained for 10 days following the expression of cocaine kindled seizures. In the brains of mice sensitized to the low dose of cocaine (15 mg/kg) no change in NMDA receptor binding was observed compared with control values. The present findings suggest the following: (a) The induction of cocaine kindling is associated with increased NMDA receptor binding activity in the striatum, amygdala and hippocampus; (b) the maintenance of cocaine kindling depends on increased NMDA receptor binding in the amygdala and hippocampus; (c) sensitization to cocaine-induced horizontal locomotor activity may be independent of elevation in NMDA receptor binding. PMID- 10665818 TI - Taurine-induced synaptic potentiation: role of calcium and interaction with LTP. AB - Taurine induces a long-lasting potentiation of excitatory synaptic potentials due to the enhancement of both synaptic efficacy and axon excitability in the CA1 area of rat hippocampal slices. In this study, we characterized the role of Ca2+ in the generation of these long-lasting taurine effects. Taurine perfusion in a free-Ca2+ medium did not induce changes in either field excitatory synaptic potentials (fEPSP) slope or fiber volley (FV) amplitude. Intracellular recordings with a micropipette filled with the Ca2+ chelator BAPTA, prevented the EPSP potentiation induced by taurine in the impaled cell, whereas a long-lasting potentiation of the simultaneously recorded fEPSP was obtained. The depletion of intracellular Ca2+ stores by thapsigargin (1 microM), an inhibitor of endosomal Ca2+-ATPase, transformed the taurine-induced potentiation into a transitory process that declined to basal values after taurine withdrawal. Taurine-induced potentiation was not significantly affected by kynurenate (glutamate receptor antagonist), or nifedipine (high-voltage-activated Ca2+ channel antagonist). But, the presence of nickel (50 microM), an antagonist of low-voltage-activated Ca2+ channel, inhibited the taurine-induced potentiation, indicating that Ca2+ influx through this type of Ca2+ channels could account for the Ca2+ requirement of the taurine-induced potentiation. Occlusion experiments between tetanus-induced long term potentiation (LTP) and taurine-induced potentiation indicate that both processes share some common mechanisms during the maintenance period. PMID- 10665819 TI - Deduction of amino acid residues in the GABA(A) receptor alpha subunits photoaffinity labeled with the benzodiazepine flunitrazepam. AB - Peptide mapping and microsequencing were used to infer the site of photoaffinity labeling by the gamma-aminobutyric acidA receptor modulator [3H]flunitrazepam. Peptide mapping with and without N-deglycosylation was used to restrict the domain for photoaffinity labeling to residues 74-123 of the bovine alpha1 subunit, in agreement with a previously predicted labeling domain between residues 59-148 based on cyanogen bromide fragmentation. Edman degradation of partially purified photolabeled peptides gave release of 3H counts in the ninth cycle of a tryptic peptide sequence. A second V8/chymotryptic peptide produced an impure sequence with release of 3H counts in the seventh through ninth cycle of sequence. The combined data support those previously reported, i.e., that the primary site for photoaffinity labeling by [3H]flunitrazepam is His102 of the bovine alpha1 subunit. In addition we also detected possible secondary labeling of Pro97. PMID- 10665820 TI - Reversal of the activity-dependent suppression of GABA-mediated inhibition in hippocampal slices from gamma-vinyl GABA (vigabatrin)-pretreated rats. AB - The antiepileptic drug, gamma-vinyl GABA (GVG, vigabatrin), is an irreversible inhibitor of GABA-transaminase, the enzyme responsible for the breakdown of GABA. In hippocampal slices prepared from rats pretreated with either an anticonvulsant dose of GVG (1500 mg/kg) or saline, electrophysiological recordings were performed in order to examine the effects of GVG pretreatment on GABAergic neurotransmission. Although GVG had no effect on the effectiveness of GABA mediated inhibition when elicited by a single stimulus, it reversed the activity dependent depression of inhibition which is typically observed when inhibitory pathways are activated repetitively by a train of stimuli delivered at low frequency. Similarly, GVG pretreatment prevented the progressive decline in the amplitude of monosynaptic inhibitory postsynaptic potentials (IPSPs) during low frequency stimulation of inhibitory interneurons. Thus, in slices from GVG pretreated rats, the amplitudes of both the fast and slow components of the last of a series of IPSPs evoked by a 5 Hz, 4 s train were maintained at 91.5 +/- 6.6% and 87.7 +/- 6.5%, respectively, compared to 61.1 +/- 3.9% and 57.1 +/- 5.0% in control slices. Finally, in slices from GVG pretreated rats, we observed a reduction in the ability of the GABA(B) receptor agonist, baclofen, to decrease the amplitude of monosynaptic inhibitory postsynaptic currents. These results suggest that GVG may produce its frequency-dependent actions by reducing the function of release regulating presynaptic GABA(B) autoreceptors. The frequency dependent reinforcement of inhibition by GVG may importantly contribute to the anticonvulsant effectiveness of this compound. PMID- 10665821 TI - Dopaminergic behaviors and signal transduction mediated through adenylate cyclase and phospholipase C pathways. AB - We determined the relative effects of chemical receptor inactivation on dopaminergic signaling through adenylate cyclase and phospholipase C pathways and evaluated the behavioral implications of such receptor manipulations. Groups of rats were given intraperitoneal injections of 10 mg/kg N-ethoxycarbonyl-2-ethoxy 1,2-dihydroquinoline (EEDQ), a reagent that differentially inactivates neurotransmitter receptors. Control and treated animals were used to assess dopaminergic-mediated behaviors or brain tissues were prepared from the animals and used to assay D1-like receptor binding and agonist-stimulated second messenger formation. EEDQ decreased by 75% the number of D1-like binding sites and completely abolished dopamine-stimulated cyclic AMP formation in striatal membranes. Conversely, dopamine-stimulated phosphoinositide hydrolysis was insensitive to inactivation by EEDQ as examined over different durations of EEDQ treatment, in different brain regions, or with different concentrations of the D1 like receptor agonist SKF38393. EEDQ-pretreated animals lost their stereotypic response to apomorphine but showed increased vacuous jaw movements in response to apomorphine or SKF38393. Basal catalepsy was increased and SCH23390 was unable to further enhance catalepsy beyond the basal levels in the lesioned animals. In naive animals, SCH23390 catalepsy was reversed by apomorphine, and apomorphine stereotypy was reversed by SCH23390. Taken together, the present results imply that the dopamine-sensitive phospholipase C system mediates a subset of dopaminergic behaviors, notably vacuous jaw movements, in contrast to stereotypy and catalepsy which appear to be respectively mediated through stimulation and inhibition of the adenylate cyclase-coupled dopaminergic system. PMID- 10665822 TI - Striatal application of nicotine, but not of lobeline, attenuates dopamine release in freely moving rats. AB - We investigated the physiological role of native low- and high-affinity nicotinic acetylcholine receptors (nAChRs) in regulating dopamine (DA) release from striatal DA terminals. To evaluate the functional interactions of the two receptor subtypes, nicotine (which interacts with both high- and low-affinity nAChRs) and lobeline (which selectively interacts with high-affinity nAChRs) were perfused through a microdialysis probe implanted into the striatum of freely moving rats. The DA content of successive dialysates was quantified by HPLC with an electrochemical detector. A short-lasting (1-min) perfusion of nicotine or lobeline dose-dependently increased the DA content of striatal dialysates. A second application of the same dose of nicotine resulted in an attenuated DA increase, compared with the increase elicited by the first application; however, the DA increase elicited by a second application of lobeline was similar to that of the first lobeline application. The nicotine-induced response was not attenuated when it followed a lobeline perfusion; in contrast, if the nicotine perfusion preceded that of lobeline, the lobeline-induced response was attenuated. In the presence of mecamylamine (a noncompetitive nAChR antagonist), the increase in DA content of striatal dialysate samples induced by either nicotine or lobeline was attenuated. However, in the presence of methyllycaconitine (a preferential antagonist for low-affinity alpha7 homomeric nAChRs), the nicotine response was attenuated but that of lobeline was unaffected. These results suggest that the functional inactivation of striatal nAChRs requires the simultaneous activation of both low- and high-affinity nAChRs. Since lobeline is devoid of reinforcing properties, one might infer that the reinforcing properties of nicotine require the simultaneous activation of high- and low-affinity brain nAChRs. PMID- 10665823 TI - Effects of tranylcypromine on thyroid hormone metabolism and concentrations in rat brain. AB - The effect of 14 days administration of the anti-depressant tranylcypromine (TCP) on iodothyronine deiodinase activities and the concentrations of thyroxine (T4) and triiodothyronine (T3) were investigated in homogenates of up to nine regions of the rat brain. The activity of the 5III deiodinase isoenzyme, which catalyses the inactivation of T3 to 3,3'-diiodothyronine (3,3'-T2), was enhanced in eight brain regions. However, the brain levels of T4 were completely unchanged and the T3 concentrations were significantly reduced in the frontal cortex only. Therefore, we also measured the T3 concentrations of three subcellular fractions (nuclei, synaptosomes and mitochondria) of six brain regions. TCP induced a significant reduction in T3 levels in the synaptosomes of the frontal cortex and significant increases in the mitochondrial T3 concentrations in the amygdala. The latter effect was replicated after 14 days administration of 5 mg/kg desipramine. No effects of either drug on nuclear concentrations of T3 were seen in any brain region. As the amygdala is critically involved in the affective coloring of sensory stimuli, the increase in T3 concentrations in the mitochondria of this brain region may be of relevance for the mechanism of action of anti-depressant drugs. PMID- 10665824 TI - Differential adaptation of brain 5-HT1A and 5-HT1B receptors and 5-HT transporter in rats treated chronically with fluoxetine. AB - Quantification of receptor binding sites and their encoding mRNAs, and electrophysiological recordings, were used to assess central serotonin (5-HT) neurotransmission in rats 24 h after a 2-3 week treatment with the selective 5-HT reuptake inhibitor fluoxetine (8 mg/kg i.p., daily). Binding studies showed that this treatment affected neither 5-HT1A nor 5-HT1B binding sites in all brain areas examined. However, a significant decrease (-38%) in 5-HT1A mRNA levels in the anterior raphe area (but not forebrain regions) and increases in 5-HT1B mRNA levels in the striatum (+127%) and the cerebral cortex (+34%) were noted in fluoxetine-treated rats. Electrophysiological recordings in brain slices showed that chronic fluoxetine treatment reduced the potency of the 5-HT1A agonist 8 hydroxy-2-(di-n-propylamino)tetralin to inhibit neuronal activity in the dorsal raphe nucleus, but did not affect 5-HT1A-evoked responses of CA1 pyramidal cells in the hippocampus. These data further demonstrate that fluoxetine-induced adaptive changes in 5-HT neurotransmission exhibit marked regional differences. The decrease in 5-HT1A mRNA levels in the anterior raphe suggests that fluoxetine induced desensitization of 5-HT1A autoreceptors involves changes at the transcription level. PMID- 10665825 TI - Immunohistochemical localisation of the 5-HT2C receptor protein in the rat CNS. AB - 5-HT2C receptor mRNA has a widespread distribution in the human and rat CNS but the absence of a specific high affinity ligand has made autoradiographic localisation of the receptor difficult. By raising polyclonal antibodies against the rat 5-HT2C receptor protein this study reports the immunohistochemical distribution of this receptor in the rat CNS. A sephadex purified 5-HT2C antiserum visualised a single immunopositive band (54 kDa) in Western blots of membranes prepared from several rat brain regions and caused intense membrane immunofluorescence in HEK 293 cells transfected with h5-HT2C cDNA, which were both attenuated by incubation with the antigenic peptide sequence (200-300 microM). 5-HT2C-like immunoreactivity was located on neurones throughout the CNS. The most abundant 5-HT2C-like immunoreactive cell bodies were in the anterior olfactory nucleus, medial and intercalated amygdaloid nuclei, hippocampus layers CA1 to CA3, laterodorsal and lateral geniculate thalamic nuclei, caudate-putamen and several areas of the cortex (including piriform and frontal), consistent with this receptor being located postsynaptic to serotonergic neurones. Immunopositive neurones were also found in the dorsal raphe, suggesting that 5-HT2C receptors may be on some serotonergic neurones. The overall distribution of 5-HT2C-like immunoreactivity complements previous findings with conventional radioligands and agrees well with reported levels of 5-HT2C receptor mRNA. PMID- 10665826 TI - The involvement of tachykinin NK2 and NK3 receptors in central sensitization of a spinal withdrawal reflex in the decerebrated, spinalized rabbit. AB - Repetitive electrical stimulation of the sural nerve at a strength sufficient to excite C-fibres results in prolonged (> 20 min) increases in the reflex responses of gastrocnemius motoneurones evoked by stimulation of myelinated axons in the sural nerve. We have tested the susceptibility of this effect to blockade of tachykinin NK2 and NK1 receptors. In the control state, iterative stimulation of sural nerve C fibres increased gastrocnemius reflexes to a peak of between 200 and 400% of pre-stimulus levels, an effect that recovered to baseline values over 23-30 min. Pre-treatment with selective antagonists for NK2 (SR 48968, 1 mg/kg i.v.) or NK3 (SR 142801, 0.1 and 1 mg/kg i.v.) receptors failed to alter the peak facilitation resulting from sural nerve stimulation. However, both drugs significantly reduced the duration of enhancement of reflexes, to 18 min after SR 48968 and to 5 min after SR 142801. The inactive enantiomers of these compounds (SR 48965 and SR 142806, both 1 mg/kg i.v.) did not reduce facilitation of reflexes after sural nerve stimulation. We conclude that activation of tachykinin NK3 receptors is a major factor in maintaining central sensitization of the sural gastrocnemius reflex after electrical stimulation of sural nerve C-fibres. The effects of SR 48968 were surprisingly weak and may have resulted from an effect of this compound at NK3 receptors. PMID- 10665827 TI - NK3 receptor blockade prevents hyperalgesia and the associated spinal cord substance P release in monoarthritic rats. AB - Previous studies in vitro have shown that NK3 receptors exist on primary afferent terminals in rat spinal cord and mediate potentiation of the depolarisation evoked substance P (SP) release. In the present study we have investigated the role of the NK3 receptor-mediated SP release system in a model of inflammatory pain. Monoarthritis was induced in rats by unilateral injection of complete Freund's adjuvant (CFA); withdrawal latencies to a thermal stimulus were subsequently measured at various times following CFA. The CFA-treated paw displayed hyperalgesia as early as 4 h after CFA injection and hyperalgesia was maintained until day 4 but had disappeared by day 21. The thermal hyperalgesia was associated with an increase in basal SP release from spinal cord synaptosomes. The possible involvement of endogenous neurokinin B acting at NK3 receptors was tested by using SB 223412-A [(S)-(-)-N-(alpha-ethylbenzyl)-3 hydroxy-2-phenylquinoline-4-carbo xamide hydrochloride], a novel, potent (Ki=30 nM) and selective (Ki>10,000 nM for NK1 and NK2 receptors), non-peptidic NK3 receptor antagonist. In vitro SB 223412-A antagonised the potentiation of SP release produced by senktide in spinal cord synaptosomes. Administered systemically to monoarthritic rats (50 mg/kg, p.o., b.i.d., for 4 days), the NK3 receptor antagonist SB 223412-A significantly reduced thermal hyperalgesia and normalised the basal release of SP from spinal cord synaptosomes. The data suggest that neurokinin B acting at NK3 receptors that mediate SP release within the spinal cord play a role in inflammation. These NK3 receptors may represent, therefore, appropriate targets in the therapy of inflammatory pain. PMID- 10665828 TI - U50,488 protection against HIV-1-related neurotoxicity: involvement of quinolinic acid suppression. AB - The pathogenesis of human immunodeficiency virus type 1 (HIV-1) encephalopathy has been associated with multiple factors including the neurotoxin quinolinate (an endogenous N-methyl-D-aspartate [NMDA] receptor ligand) and viral proteins. The kappa opioid receptor (KOR) agonist U50,488 recently has been shown to inhibit HIV-1 p24 antigen production in acutely infected microglial cell cultures. Using primary human brain cell cultures in the present study, we found that U50,488 also suppressed in a dose-dependent manner the neurotoxicity mediated by supernatants derived from HIV-1-infected microglia. This neuroprotective effect of U50,488 was blocked by the KOR selective antagonist nor binaltorphimine. The neurotoxic activity of the supernatants from HIV-1-infected microglia was blocked by the NMDA receptor antagonists 2-amino-5 phosphonovalerate and MK-801. HIV-1 infection of microglial cell cultures induced the release of quinolinate, and U50,488 dose-dependently suppressed quinolinate release by infected microglial cell cultures with a corresponding inhibition of HIV-1 p24 antigen levels. These findings suggest that the kappa opioid ligand U50,488 may have therapeutic potential in HIV-1 encephalopathy by attenuating microglial cell production of the neurotoxin quinolinate and viral proteins. PMID- 10665829 TI - The role of new anesthetic agents. AB - The three anesthetic drugs introduced most recently to the market are sevoflurane, desflurane, and ropivacaine. Sevoflurane and desflurane are both inhalational anesthetic agents and ropivacaine is a local anesthetic agent. Sevoflurane provides a rapid onset and offset of action; it is well tolerated with little airway irritation. It is hemodynamically stable, with low potential for toxicity. Concerns about its interaction with soda lime during low-flow anesthesia with the production of Compound A have not proved to be a clinical problem. While desflurane also provides rapid onset and recovery from anesthesia, it is not as hemodynamically stable as sevoflurane, and also causes airway irritation. Ropivacaine is a unique local anesthetic in that it is supplied as the pure S-enantiomer. It is at least as effective as bupivacaine, with lower toxicity and less motor block for the same degree of sensory block. PMID- 10665830 TI - Vascular sodium pump: endothelial modulation and alterations in some pathological processes and aging. AB - The vascular Na+ pump maintains intracellular ionic concentration and controls membrane potential. Its inhibition by cardiac glycosides enhances the intracellular Na+ concentration. This in turn activates the Na+-Ca2+ exchange mechanism, which induces intracellular Ca2+ increase, membrane depolarization, and noradrenaline release from perivascular adrenergic nerve endings; mechanisms that promote vasoconstriction. This article reviews the relevance of the Na+ pump in vascular tone regulation and the modulation of its activity by the endothelium. The endothelium negatively modulates the vasoconstriction elicited by Na+ pump inhibition by the release of nitric oxide, according to some authors, or an unknown factor, as suggested by others. The possible existence of endogenous digitalis-like factors is also reviewed, as is the involvement of the vascular Na+ pump in some cardiovascular disorders and aging. PMID- 10665831 TI - General overview of mineralocorticoid hormone action. AB - The adrenal cortex elaborates two major groups of steroids that have been arbitrarily classified as glucocorticoids and mineralocorticoids, despite the fact that carbohydrate metabolism is intimately linked to mineral balance in mammals. In fact, glucocorticoids assured both of these functions in all living cells, animal and photosynthetic, prior to the appearance of aldosterone in teleosts at the dawn of terrestrial colonization. The evolutionary drive for a hormone specifically designed for hydromineral regulation led to zonation for the conversion of 18-hydroxycorticosterone into aldosterone through the catalytic action of a synthase in the secluded compartment of the adrenal zona glomerulosa. Corticoid hormones exert their physiological action by binding to receptors that belong to a transcription factor superfamily, which also includes some of the proteins regulating steroid synthesis. Steroids stimulate sodium absorption by the activation and/or de novo synthesis of the ion-gated, amiloride-sensitive sodium channel in the apical membrane and that of the Na+/K+-ATPase in the basolateral membrane. Receptors, channels, and pumps apparently are linked to the cytoskeleton and are further regulated variously by methylation, phosphorylation, ubiquination, and glycosylation, suggesting a complex system of control at multiple checkpoints. Mutations in genes for many of these different proteins have been described and are known to cause clinical disease. PMID- 10665832 TI - New approaches to rational drug design. AB - Rational drug design has emerged as a powerful technique. In this review, three new developments in rational drug design are explored. These developments include new methods to find binding sites for small molecules on the surface of a protein, the suggestion that the protein environment may change the shape of a protein sufficiently to alter drug design, and the use of data emerging from structural genomics in drug design. Although these are three new and distinct areas, the insights derived from these studies suggest a reason for the observation that similar drugs do not always bind to a protein in the same manner. PMID- 10665833 TI - The contribution of electrophysiology to knowledge of the acute and chronic effects of ethanol. AB - This review describes the effects of ethanol on the components of neuronal transmission and the relationship of such effects to the behavioural actions of ethanol. The concentrations of ethanol with acute actions on voltage-sensitive ion channels are first described, then the actions of ethanol on ligand-gated ion channels, including those controlled by cholinergic receptors, 5 hydroxytryptamine receptors, the various excitatory amino acid receptors, and gamma-aminobutyric acid receptors. Acute effects of ethanol are then described on brain areas thought to be involved in arousal and attention, the reinforcing effects of ethanol, the production of euphoria, the actions of ethanol on motor control, and the amnesic effects of ethanol; the acute effects of ethanol demonstrated by EEG studies are also discussed. Chronic effects of alcohol on neuronal transmission are described in the context of the various components of the ethanol withdrawal syndrome, withdrawal hyperexcitability, dysphoria and anhedonia, withdrawal anxiety, craving, and relapse drinking. Electrophysiological studies on the genetic influences on the effects of ethanol are discussed, particularly the acute actions of ethanol and electrophysiological differences reported in individuals predisposed to alcoholism. The conclusion notes the concentration of studies on the classical transmitters, with relative neglect of the effects of ethanol on peptides and on neuronal interactions between brain areas and integrated patterns of neuronal activity. PMID- 10665834 TI - Superoxide in the pulmonary circulation. AB - Superoxide formation in pulmonary tissue is modulated by cytokines, PO2, shear force, and disease states, and can be stimulated by drugs. Superoxide has diverse actions on pulmonary cells, including smooth muscle contraction, interaction with redox enzymes, cell proliferation, and gene transcription. In the lungs, there is an impressive array of specific defence mechanisms that destroy superoxide, especially superoxide dismutase (SOD) and metallothionein. Superoxide formation is increased in hyperoxia (e.g., oxygen therapy); however, superoxide-forming enzymes also can be up-regulated in hypoxia. Superoxide has been implicated in acute respiratory distress syndrome, lung ischaemia-reperfusion injury, and lung transplantation. Novel approaches to therapy have been explored, including SOD gene therapy and SOD targeting to the lung. In the future, new drugs interacting with superoxide may provide significant advances in the treatment of lung diseases. PMID- 10665835 TI - Unusual transcriptional and translational regulation of the bacteriophage Mu mom operon. AB - The bacteriophage Mu mom gene encodes a novel DNA modification that protects the viral genome against a wide variety of restriction endonucleases. Expression of mom is subject to a series of unusual regulatory controls. Transcription requires the action of a phage-encoded protein, C, which binds (probably as a dimer) the mom promoter from -33 to -52 (with respect to the transcription start site) in two adjacent DNA major grooves on one face of the helix. No apparent direct interaction between C and the host RNA polymerase (RNAP) is evident; however, C binding alters mom DNA conformation. In the absence of C, RNAP binds the mom promoter at a site that results in transcription in a direction away from the mom gene. The function of this transcription is unknown. An additional layer of transcriptional regulation complexity is due to the fact that the host Dam DNA (N6-adenine)methyltransferase is required. Dam methylation of three closely spaced upstream GATC sequences is necessary to prevent binding by the host protein, OxyR, which acts as a repressor. Repression is not mediated by inhibition of C binding, but rather through interference with C-mediated recruitment of RNAP to the correct site. Translation of mom is regulated by the phage Com protein. Com is only 62 amino acids long and contains a zinc finger like structure (coordinated by four cysteine residues) in the amino terminal domain. Com binds mom mRNA 5' to the mom open reading frame, whose translation start signals are contained in a stem-loop translation-inhibition-structure. Com binding to its target site (5' to and adjacent to the translation-inhibition structure) results in a stable change in RNA secondary structure that exposes the translation start signals. PMID- 10665836 TI - DNA methylation analysis: a review of current methodologies. AB - The relationship between levels of DNA methylation and gene activity has been known for some time. Many of the early procedures developed gave only somewhat limited information about methylation patterns, for example, the total level of 5 methyl cytosine in the genome or the frequency of methylation of cytosines within certain restriction sites. However, in the last few years, there has been an explosion of interest in DNA methylation, and with it, many new and powerful techniques have been developed to facilitate its study. In this paper, the key techniques currently available are reviewed and the advantages, disadvantages, and potential artifacts of each are discussed. PMID- 10665837 TI - Nitric oxide in the lung: therapeutic and cellular mechanisms of action. AB - Nitric oxide is produced by many cell types in the lung and plays an important physiologic role in the regulation of pulmonary vasomotor tone by several known mechanisms. Nitric oxide stimulates soluble guanylyl cyclase, resulting in increased levels of cyclic GMP in lung smooth muscle cells. The gating of K+ and Ca2+ channels by cyclic GMP binding is thought to play a role in nitric oxide mediated vasodilation. Nitric oxide may also regulate pulmonary vasodilation by direct activation of K+ channels or by modulating the expression and activity of angiotensin II receptors. Administration of nitric oxide by inhalation has been shown to acutely improve hypoxemia associated with pulmonary hypertension in humans and animals. This is presumably due to its ability to induce pulmonary vasodilation. Inhaled nitric oxide improves oxygenation and reduces the need for extracorporeal membrane oxygenation in term and near-term infants with persistent pulmonary hypertension. However, long-term benefits to these infants have been difficult to demonstrate. In other pathologic conditions, such as prematurity and acute respiratory distress syndrome, short-term benefits have not been shown conclusively to outweigh potential toxicities. For example, high-dose inhaled nitric oxide decreases surfactant function in the lung. Inhaled nitric oxide also acts as a pulmonary irritant, causing priming of lung macrophages and oxidative damage to lung epithelial cells. Conversely, protective effects of nitric oxide have been described in a number of pathological states, including hyperoxic and ischemia/reperfusion injury. Nitric oxide has also been reported to protect against oxidative damage induced by other reactive intermediates, including superoxide anion and hydroxyl radical. The dose and timing of nitric oxide administration needs to be ascertained in clinical trials before recommendations can be made regarding its optimal use in patients. PMID- 10665838 TI - New insights into the pharmacodynamic and pharmacokinetic properties of statins. AB - The beneficial effects of statins are assumed to result from their ability to reduce cholesterol biosynthesis. However, because mevalonic acid is the precursor not only of cholesterol, but also of many nonsteroidal isoprenoid compounds, inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase may result in pleiotropic effects. It has been shown that several statins decrease smooth muscle cell migration and proliferation and that sera from fluvastatin-treated patients interfere with its proliferation. Cholesterol accumulation in macrophages can be inhibited by different statins, while both fluvastatin and simvastatin inhibit secretion of metalloproteinases by human monocyte-derived macrophages. The antiatherosclerotic effects of statins may be achieved by modifying hypercholesterolemia and the arterial wall environment as well. Although statins rarely have severe adverse effects, interactions with other drugs deserve attention. Simvastatin, lovastatin, cerivastatin, and atorvastatin are biotransformed in the liver primarily by cytochrome P450-3A4, and are susceptible to drug interactions when co-administered with potential inhibitors of this enzyme. Indeed, pharmacokinetic interactions (e.g., increased bioavailability), myositis, and rhabdomyolysis have been reported following concurrent use of simvastatin or lovastatin and cyclosporine A, mibefradil, or nefazodone. In contrast, fluvastatin (mainly metabolized by cytochrome P450-2C9) and pravastatin (eliminated by other metabolic routes) are less subject to this interaction. Nevertheless, a 5- to 23-fold increase in pravastatin bioavailability has been reported in the presence of cyclosporine A. In summary, statins may have direct effects on the arterial wall, which may contribute to their antiatherosclerotic actions. Furthermore, some statins may have lower adverse drug interaction potential than others, which is an important determinant of safety during long-term therapy. PMID- 10665839 TI - Fetal hepatic drug elimination. AB - The majority of studies of fetal hepatic elimination have concentrated on the expression and activity of the metabolizing enzymes, but the unique physiologic milieu of the fetal liver should also be considered. The basic structure of the liver is formed by the end of the first trimester. The fetal hepatic circulation differs substantially from that of the adult in that there is an extra input vessel, the umbilical vein, and there is shunting of 30-70% of hepatic blood flow via the ductus venosus. The left and right lobes of the fetal liver seem to function independently with respect to a variety of biochemical parameters, due at least in part to the lower oxygen supply to the right lobe. The zonation of drug-metabolizing enzymes along the hepatic acinus, which is prominent in the adult liver, is absent in the fetal liver. Unlike rodent species, the human fetal liver has a significant capacity for drug metabolism. Of the oxidative enzymes, CYP3A7 accounts for up to 50% of total fetal hepatic cytochrome P450 content. Expression of this enzyme decreases dramatically after birth. CYP1A1 and CYP2D6 have also been detected in human fetal liver, but whether CYP2E1 is expressed remains controversial. Several other cytochrome P450s have been identified and await characterization. Fetal hepatic drug conjugation may prolong fetal exposure to the metabolites produced, which, being more water soluble, do not readily cross the placenta back to the mother and, if excreted in fetal urine, can be recycled in the fetus via amniotic fluid and fetal swallowing. Limited activity of glucuronidation enzymes has been demonstrated in human fetal liver in contrast to the activity of sulfation enzymes, which is significant. Limited in vivo studies in fetal sheep have demonstrated significant fetal hepatic drug elimination, and this has been confirmed in studies of the isolated perfused fetal sheep liver. Our understanding of fetal hepatic elimination processes has advanced steadily over the years. Future developments, however, should consider more fully the influence of the unique physiological milieu of the fetal liver, in addition to the expression and activity of drug metabolizing enzymes. PMID- 10665840 TI - Tumor necrosis factor-alpha: molecular-biological aspects minireview. AB - Tumor necrosis factor-alpha (TNF-alpha) a proinflammatory cytokine with multiple actions was first identified for its anticancer activity. However, TNF-alpha has a beneficial function in activation of host defense, its uncontrolled production can lead to pathological consequences. At the cellular level, it is able to exert obviously opposing effects: apoptosis and activation. It modulates survival and activates genes through various intermediates, including protein kinases, protein phosphatases, reactive oxygen intermediates, phospholipases, proteases, sphingomyelinases and transcription factors. In this review, the INF-alpha is characterized at the molecular and cellular level (TNF-alpha mediated signal transduction is discussed in the first part, regulation of its expression in the second one), as well as methods of its determination in biological materials, giving special emphasis to the molecular-biological approach. The full understanding of the molecular mechanism of TNF-alpha will provide the basis for a pharmacological approach intended to inhibit or potentiate selected biological actions of this cytokine. PMID- 10665841 TI - Construction and testing of gene therapy retroviral vector expressing bacterial cytosine deaminase gene. AB - A retroviral vector containing gene for bacterial enzyme cytosine deaminase (CD) under the control of viral LTR sequences was constructed and transfected into packaging cell line GP+envAm12. High virus titer producing single cell clone (1 x 10(7) cfu/ml, determined on NIH 3T3 cells) was isolated and used to transfer CD gene into human mammary carcinoma cell lines in vitro. Transduced cells exhibited high sensitivity to the antifungal drug 5-fluorocytosine (5-FC), whereas parental cells did not. Cocultivation of CD-positive and CD-negative parental cells showed bystander effect, dependent on the ratio of CD-positive cells. No enhancement of 5-FC cytotoxicity by leucovorin was observed in cells expressing cytosine deaminase. PMID- 10665842 TI - Aberrant markers expression in T- and B-lymphoid and myeloid leukemia cells of different differentiation stages. AB - The aim of the study was to ascertain if in T acute lymphoblastic leukemia (T ALL), B acute lymphoblastic leukemia (B-ALL) and acute myeloid leukemia (AML) of different differentiation stages the coexistence of aberrant markers correlate with the degree of leukemic blasts maturation. We evaluated the results of surface and intracellular markers in 42 T-ALL, 86 B-ALL and 71 AML cases. A large panel of monoclonal antibodies (MoAbs) against T-cell, B-cell, myeloid cell and non-lineage specific structures has been used. Patients had dual-color flow cytometric immunophenotyping performed by FACStar flow cytometer. The correct immunological diagnosis of followed new cases before any treatment has been performed and simultaneously the presence of atypical/aberrant phenotypes has been studied and correlated with leukemia cells differentiation stage. A great deal of T-ALL and AML, in opposite to B-ALL cases, revealed a high proportion of atypical phenotypes (55, 75 and 36%, respectively), which are absent in nonleukemic cells. We found out that these atypical phenotypes were present in T ALL, AML (not clearly in B-ALL) through all differentiation stages and so we obtained an evidence that they might represent an abnormal/atypical rather than an immature phenotype, as it was postulated till now by several authors. PMID- 10665843 TI - Angiogenesis inhibitor TNP-470: cytotoxic effects on human neoplastic cell lines. AB - Angiostatic substance TNP-470 displayed moderate cytotoxicity towards human leukemia HL-60, HL-60/ADR, HL-60/VCR and myeloma ARH77 cell lines with IC50 in the range 5-10 microM of concentrations and slightly higher IC50 for myeloma cell line U266. IC50 for ovarian CH-1, A2780 and A2780/ADR cell lines was in the range 10-15 microM with the exception of platinum-resistant SKOV3 cell line (more than 40 microM ). The IC50 values for MDA-MB-231 and MCF-7 breast carcinoma cell lines were 15 and 25 microM, respectively. In human hemopoietic neoplastic cell lines examined, TNP-470 induced the appearance of subpopulation with sub-G0 DNA content, suggesting the apoptosis-inducing potential of TNP-470 in these cells. No TNP-470-induced drug uptake modulation in drug-resistant leukemia cell line HL 60/VCR was observed. TNP-470 induced accumulation of cells in G0/G1 phase of cell cycle. There was no TNP-470-induced inhibition of MMP collagenase activity or MMP (MMP2 and MMP9) production in the human fibrosarcoma cells HT 1080 in vitro. PMID- 10665844 TI - Characterization of APC exon 15 germ-line mutation in FAP family with severe phenotype showing extracolonic symptoms. AB - The adenomatous polyposis coli (APC) gene plays a crucial role in colorectal carcinogenesis. Germ-line mutations of APC gene give rise to familial adenomatous polyposis coli (FAP) - autosomal dominant syndrome manifesting hundreds to thousands of colorectal polyps, if untreated with malignant progression. We have used the techniques of heteroduplex analysis (HDA), protein truncation test (PTT), single strand conformation polymorphism (SSCP) and DNA sequencing for the identification and detailed positional analysis of mutations in IFAP family with the expressive phenotype characterized by polyposis and extracolonic lesions. Detailed analysis revealed a 5bp deletion in a mutation cluster region (MCR) in exon 15 of APC gene in codon 1308. Two screened members of the FAP family exhibited this novel mutation. PMID- 10665845 TI - A note on nucleoli in granulocytic precursors of the granulopoietic proliferating compartment in patients suffering from the chronic phase of chronic myeloid leukemia treated with two different drugs with different mode of action. AB - The incidence of main nucleolar types in granulocytic precursors was studied in the granulopoietic proliferating compartment (GPC) of patients suffering from chronic phase of the chronic myeloid leukemia (CML) who were treated by the widely used therapy with two different drugs with different mode of action - hydroxyurea (HU) and interferon alpha (IFN-alpha). In comparison with IFN early stages of GPC, i.e. myeloblasts and promyelocytes in patients treated with HU possessed more frequently micronucleoli which are known to reflect the direct as well as indirect inhibition of the nucleolar biosynthetic activities. On the other hand, the incidence of micronucleoli in these cells of a small percentage of patients treated with IFN also reached the average values of these nucleoli which were noted in patients treated with HU. PMID- 10665846 TI - Expression of CD26 and DPP IV in T-acute lymphoblastic leukemia: comparison of immunocytochemistry with enzyme cytochemistry. AB - Tile possible identity ofdipeptidyl peptidase IV (DPP IV) enzymatic activity and CD26 antigen expression in phenotypically defined T-acute lymphoblastic leukemia cells (T-ALL) was examined. For comparative studies, the combination of immunocytochemistry and enzyme cytochemistry methods was used. T he strong correlation between the CD26 antigen expression and DPP IV positivity in the majority of T-lymphoblasts in T-ALL patients was evident. No CD26 antigen was expressed on DPP IV negative T-cells. The variable CD4 and/or CD8 antigen expression, frequent CD5 and CD7 positivity and absence of surface membrane CD3 antigen were the characteristic immunophenotypic features of CD26/DPP IV positive T-lymphoblasts. Moreover, the clear CD71 and CD26/DPP IV coexpression suggested the association of CD26/DPP IV positive cells with proliferation. The immunophenotype of CD26/DPP IV positive T-lymphoblasts seems to be characteristic for the relative immature cell population. In addition, noteworthy was the slight disassociation between the very high CD26 antigen expression and moderate DPP IV activity in cells of some T-ALL patients. The possible existence of enzymatically inactive structures of CD26 antigen or inactive precursors of DPP IV detectable only by immunocytochemistry was discussed. Our study indicates that CD26 antigen expression is tended to identify cells with DPP IV enzymatic activity in T-ALL patients. The results provide some more information of CD26 antigen involvement in the pathology of leukemic cells via its DPP IV enzyme activity. PMID- 10665847 TI - Comparison of antileukemic immunity against U937 cells in atopic asthmatics versus healthy controls. AB - To assess the antitumor effects in atopic asthmatics versus healthy adults, we designed this study using in vitro mononuclear cells (MNC) culture as an immunity model with human leukemic U937 cells as the target. MNCs were collected from asthmatic subjects and healthy controls. Conditioned media from the MNC cultures (MNC-CM) were collected after stimulation with various concentrations of phytohemagglutinin (PHA). We treated U937 cells with these MNC-CMs, then assayed their proliferation and differentiation after 5 days of culture. At lower PHA doses (1.25 microg/ml), as well as in absence of PHA, the asthmatic MNC-CMs inhibited U937 cells growth to a slightly greater extent than did the MNC-CMs from controls. In contrast, when higher doses of PHA were used (5, 10 microg/ml), this growth-inhibiting effect was dramatically reversed. The dual effect of MNC CM in these two groups was also shown in U937 cell differentiation assay, assessed as follows: morphological change by Liu's staining, functional change by NBT reduction test and CD 14 expression by flow cytometric detection. We suggest that the antileukemic effects of MNCs from asthmatic patients result from a slightly immunopotentiated status. This immunity may be dramatically reversed, however, after marked activation of MNCs. PMID- 10665848 TI - Metastasis as the first sign of thyroid cancer. AB - The aim of this paper is to review our experience with patients who presented with a metastatic tumor in the lymph nodes or other organs as the first sign of thyroid cancer. In 1974-1998, 18 602 patients were operated on due to goitre. There were 975 (5.2%) patients with thyroid malignant neoplasms. The group comprised 449 (46.1%) patients with papillary carcinoma, 309 (31.7%) with follicular carcinoma, 54 (5.5%) with medullary carcinoma, 106 (10.9%) with anaplastic carcinoma, and 57 (5.8%) with other types of thyroid malignant neoplasms. Out of these 975 patients, thyroid cancer was diagnosed on the basis of the detection of a metastatic tumor in 26 (2.7%) patients. In 16 (61.5%) of these patients the metastatic tumor was located in the regional lymph nodes. In 10 (38.5%) patients distant metastasis beyond the regional lymph nodes was the first sign of thyroid cancer. In (50%) patients metastasis was located in the bones, in 2 (20%) in the lung, in 1 (10%) in the heart, in 1 (10%) in the buttock, and in 1 (10%) in a central neck cyst. Metastasis was the initial manifestation of thyroid cancer in 18 (4%) of 449 papillary carcinoma patients, in 6 of 309 (1.9%) follicular carcinoma patients, and in 2 (3.7%) of 54 medullary carcinoma patients. Lymph node metastasis was the first sign of thyroid cancer in 13 (2.9%) patients with papillary carcinoma, 1 (0.3%) patients with follicular carcinoma and in 2 (3.7%) medullary carcinoma patients, and distant metastasis in 5 (1.1%) patients with papillary carcinoma and in 5 (1.6%) patients with follicular carcinoma. After the detection of the primary focus of thyroid cancer total thyroidectomy and modified neck dissection were performed in all patients. Differentiated thyroid carcinoma patients were treated complementarily with 131I and TSH suppressive doses of L-thyroxine, and medullary cancer patients with teleradiotherapy and substitutive doses of L-thyroxine. PMID- 10665849 TI - Osteoblasts and osteoclasts in bone marrow smears of cancer patients. AB - Osteoblasts and osteoclasts are the unique cells occurring in bone marrow smears in situations with high bone metabolic turnover (children, trauma, rachitis, Paget disease or tumors). The collection of 2706 sternal or iliac crest aspirates from patients with hematologic malignancies and solid tumors are presented. We demonstrated significantly higher positivity for osteoblasts-osteoclasts presentation in bone marrow smears for hematological malignancies (p < 0.05), solid tumors (p < 0.01), and especially breast cancer (p < 0.001). We found a significant association between osteoblast-osteoclast positivity and dissemination of breast cancer (p < 0.05). None of the breast cancer patients without signs of dissemination (X-ray, sonography or scintigraphy) had positivity for osteoblasts or osteoclasts. We suppose that the osteoblast-osteoclast positivity in bone marrow smears can serve as a cheap marker for breast cancer dissemination. PMID- 10665850 TI - Clodronate in the management of bone metastases: a clinical study of 91 patients. AB - The aim of our study was to evaluate the efficacy of oral clodronate supplementing systemic therapy and/or palliative irradiation in 91 patients with painful bone metastases. Clodronate was administered at a daily dose of 1600-3200 mg for a median of 11 months (range 3--36 months). Partial or complete pain relief was achieved in 61 of 88 evaluable patients (69%). Response rate to clodronate in patients who additionally received palliative bone radiation was similar to that in patients who did not receive irradiation (68 and 71%, respectively). Eleven out of 12 bed-ridden patients with metastatic bone pain regained the ability of walking after the treatment with clodronate. Bone pain relief lasted from 1.5 to 36 months (mean 9.3 months). Clodronate was well tolerated in all but three cases (3%) in whom the treatment was discontinued due to intensive adverse gastrointestinal effects. In conclusion, we observed satisfactory symptomatic effect and low rate of adverse reactions in patients with metastatic bone lesions treated with oral clodronate. Further large controlled studies with thorough patient monitoring are warranted to evaluate the real benefit of clodronate, and to define its optimal scheduling. PMID- 10665851 TI - Quantitative analysis of modes of invasion and lymph node metastases in oral squamous cell carcinoma. AB - The mode of tumor invasion has been suggested to have a relationship to the occurrence of cervical metastasis and to prognosis in oral squamous cell carcinoma (OSCC). However, a tumor usually does not have a single mode of invasion, and the importance, if any, of the relative proportions of different modes for metastatic potential has not been studied. Forty two cases of OSCC resected with cervical lymph nodes were selected, 20 of which had nodal metastases and 22 which had not. The mode of invasion in the tumor-host interface was classified as: I - pushing borders, II - bands, III - thin cords, IV - single cells and analyzed in 20 consecutive medium power fields. Also studied were other morphological parameters: perineural and angiolymphatic invasion, tissue eosinophilia, mitosis and intensity of inflammatory infiltrate at the tumor-host interface. The majority of the cases (95.2%) showed two or more modes of invasion. Modes I, II and III occurred with similar frequency in cases with and without metastases. Mode II was the commonest and most extensive in both groups. No mode of invasion was significantly associated with metastases, independent of its extension. The other morphological parameters were neither significantly associated with cervical metastasis. In conclusion, OSCC usually shows two or more modes of tumor invasion if a large extension of tumor-host interface is analyzed. However, the relative proportions of the modes have no correlation with the metastatic potential. PMID- 10665852 TI - Cancer prevalence in Italian cancer registry areas: the ITAPREVAL study. ITAPREVAL Working Group. AB - AIM: To present data on cancer prevalence for the areas covered by Italian cancer registries, by using a standardized set of data collection and elaboration criteria, and a single method of data analysis. SUBJECTS AND METHODS: Data on over 250,000 patients with cancer, diagnosed between 1978 and 1992, from 11 Italian cancer registries covering about 12% of the Italian population were collected, validated and analyzed according to the unified protocol of the ITAPREVAL project. The method implemented in the PREVAL computer program was used to provide prevalence estimates for the period covered by cancer registration. The total prevalence for each registry and for the pool of all registries was then estimated by correcting for incomplete observations due to the period in which the registration was not yet activated. All prevalence estimates were for 1992. RESULTS: Prevalence figures are presented by cancer site, age, sex, years from diagnosis and registry area. For all malignancies combined, total prevalence ranged from 1,350 per 100,000 inhabitants in Ragusa to 3,650 per 100,000 inhabitants in Romagna, the ratio between these two extremes being 2.7. For the pool of the areas covered by registration cancer prevalence was 3,100 per 100,000 females and 2,250 per 100,000 males. About a third of the total female cases and about half the male cases were diagnosed in the previous five years. Among those aged over 75 years, total prevalence was higher for males than for females: 11,300 versus 8,900 per 100,000 respectively. CONCLUSIONS: This is the first large-scale estimate of the burden of cancer in Italy. It is also one of the first studies in the world which was aimed to study cancer prevalence in detail. These data are necessary for predicting health service needs and help in the evaluation of differences in health service demand by sex, age and Italian regions. PMID- 10665853 TI - Correcting the completeness bias of observed prevalence. AB - AIMS AND BACKGROUND: The cancer prevalence in given areas can be estimated on the basis of data supplied by cancer registries. As the obtained estimate of prevalence depends on the length of the cancer registry's observation period, it is generally lower than the total prevalence in the considered area. In the present work we propose a method to calculate a correction factor of this bias in order to obtain an approximation to the total prevalence. METHODS & STUDY DESIGN: The method is based on the relationship between relative survival and incidence by age for a specific cancer site. RESULTS AND CONCLUSIONS: We provide values of the correction factor, the completeness index R, relative to the most important cancer sites, for specific ages and periods of observation of the cancer registries in Italy. In addition, we provide indications for extended use of the index when substantial variations from the basic pattern of relative survival are observed in practical situations. Furthermore, we give helpful suggestions to obtain approximate values of the correction factor to be used for ages and periods of observation that are intermediate between the ones presented in this paper. PMID- 10665854 TI - The prevalence of tumors of the breast and female genital tract in Italy. AB - Data from 10 Italian population-based cancer registries were used to estimate the prevalence of female tumors of the breast and genital tract. The total prevalence, expressed in number per 100,000, was highest for breast cancer (1,117), followed by cancer of the corpus (264) and cervix uteri (146), ovary (110), and vagina and vulva (23). For all tumors the prevalence increased with age at diagnosis. The cancer prevalence was divided into intervals from diagnosis, expressing different health needs in terms of therapy and intensity of clinical follow-up. For all tumors considered, 1-year prevalence was higher than 1-2-year prevalence, reflecting a high death risk due to perioperative mortality and to the proportion of patients diagnosed at advanced stages. The prevalence decreased in the following intervals considered. Noticeable geographic variability was observed in the prevalence across Italy, with higher proportions being registered in the northern-central regions than in the South. The two extreme 0-5-year prevalence figures (per 100,000) were: for breast cancer 568 (Genova) and 259 (Ragusa); for corpus uteri cancer 94 (Romagna) and 21 (Latina); for cervix uteri cancer 63 (Romagna) and 26 (Latina); for ovarian cancer 49 (Parma) and 21 (Latina); for cancer of the vagina and vulva 17 (Genova) and 5 (Ragusa). This variability depends mainly on incidence and on the proportion of elderly in the general population. From 1987 to 1992 there was an increase in the prevalence of tumors of the breast, ovary and vagina and vulva, especially in the elderly. The prevalence of corpus uteri cancer decreased slightly in the elderly only, whereas that of cervix uteri cancer decreased at all ages. PMID- 10665855 TI - The prevalence of colorectal cancer in Italy. AB - AIMS: To analyze the prevalence of colorectal cancer (CRC) in different areas of Italy by age, interval since diagnosis and disease stage at diagnosis, and to estimate the prevalence of CRC. These data provide estimates of patient demand on health resources. PATIENTS AND METHODS: Eleven Italian cancer registries (CRs) provided data on 33,740 patients observed for up to 15 years. For the 1,829 cases from the specialized colorectal cancer registry of Modena we analyzed prevalence by Dukes' stage and family history. PREVAL software produced observed prevalence figures by time from diagnosis; to determine the total prevalence, correction factors were applied to the observed data. RESULTS: At the end of 1992, five-year CRC prevalence was high (close to 200 per 100,000) in Genova, Parma, Romagna and Firenze, and low (around 75 per 100,000) in the southern areas of Latina and Ragusa. For all CRs, 86 patients per 100,000 population were alive up to 2 years from diagnosis and 77 per 100,000 between 2 and 5 years from diagnosis. The 5 year prevalence of patients diagnosed with Dukes' B or C (high risk of recurrence and requiring postoperative surveillance) was 152 per 100,000; that of Dukes' A patients 36 per 100,000 (considered cured after surgery and not requiring intensive follow-up or care); that of unstaged patients plus those with distant metastasis at diagnosis was 28 per 100,000 at 5 years (requiring palliative care but not follow-up). The 12-year prevalence of HNPCC was 23 per 100,000, or about 7% of the total; for such patients knowledge of the long-term prevalence is important because they are diagnosed young and are at high risk of multiple tumor development. CONCLUSIONS: 70% of the prevalent patients diagnosed within 5 years prior to the prevalence date were likely to require care for cancer recurrence, while 13% of the prevalent cases required care for distant metastases. PMID- 10665856 TI - Cancer prevalence in the elderly. ITAPREVAL Working Group. AB - AIMS AND BACKGROUND: To describe the cancer prevalence in elderly Italian people and analyze the differences, if any, with the prevalence among younger subjects. METHODS & STUDY DESIGN: The cancer prevalence among elderly patients (65 years and over), the three age classes encompassing elderly age (65-74 years, 75-84 years, 85 years and over) and younger patients (0-64 years) was computed using the PREVAL method on the basis of the incident cases over the period 1976-1992 followed up to 31 December 1992 (prevalence reference date). Data were collected by 11 Italian cancer registries. RESULTS: The observed prevalence figures for all cancers (except skin epitheliomas), both sexes combined and considering the whole elderly group, were 1,090 and 3,601 cases per 100,000 one and five years since diagnosis, respectively; the prevalence increased up to the 75-84 age group and showed a slight decrease after age 85. With regard to specific cancer sites, in men bladder and prostate had the highest prevalence 5 years from diagnosis (more than 800 cases per 100,000), followed by colon and lung (about 500 cases per 100,000) stomach and rectum (about 300 cases per 100,000); in women breast cancer ranked first (more than 1,000 cases per 100,000), followed by colon (about 350 cases per 100,000), corpus uteri, stomach and rectum cancers (between 150 and 200 cases per 100,000). For all malignancies and the two sexes combined the prevalence figures were about six times higher in the older than in the younger age group. CONCLUSIONS: These figures confirm the important role of aging in determining the increase in cancer prevalence. The resulting prevalence figures clearly indicate the cancer burden placed on health care services; moreover, the figures will probably increase in the next decades due to a possible improvement in survival and to the dramatic aging of the population, assuming a stable trend for incidence rates. This picture will represent a major challenge for politicians and those dealing with health care planning and social policies in general, especially in the light of the reduction of the available financial resources and the specific features of medical and social needs in the elderly. PMID- 10665857 TI - Cancer prevalence in Italian regions with local cancer registries. AB - OBJECTIVE: To provide estimates and projections of cancer incidence and prevalence for those Italian regions whose population is partially covered by a cancer registry (CR) and to determine to what extent local CRs can be considered representative of the region, thus improving the potential of the information provided by CRs. METHODS: A statistical method, MIAMOD (mortality-incidence analysis model), was used to estimate regional cancer incidence and prevalence from regional cancer mortality data and patient survival data recorded by the cancer registries. Estimates of the cancer incidence and prevalence in the various regions have thus been obtained for a number of major cancer sitas. A first and important step in validating the regional estimates has been the comparison of the MIAMOD estimates in the areas covered by the cancer registries with empirical incidence and prevalence observed by CRs, in order to assess the consistency in data, methods and assumptions. Empirical prevalence has been calculated by counting patients with a diagnosis of cancer who were alive on the reference date by PREVAL method. A correction factor has been applied to include patients diagnosed before the period of activity of the registry. RESULTS: General consistency was found between empirical and estimated (by MIAMOD) incidence and prevalence in the registry areas, which is indicative of the quality and the completeness of all data involved as well as the appropriateness of model choices. The prevalence of all cancers combined for Italian regions with CRs was estimated and projected to the year 2000 as ranging between 1,240 per 100,000 in Sicilia and 2,781 in Emilia-Romagna for men, while for women these figures were 1,765 in Sicilia and 4,019 in Liguria. Comparison of cancer prevalence in CR areas with regional estimates shows quite good consistency for Piemonte, Liguria and Lombardia, which means that the local CRs (of Torino, Genova and Varese, respectively) are representative of their respective regions. Prevalence in Emilia-Romagna appears to be rather well represented by only one, the Parma CR, of the three local CRs. The southern Italian registries of Latina and Ragusa recorded a lower cancer prevalence than was actually estimated in their respective regions. DISCUSSION: Cancer registries with a longer period of activity showed better agreement between empirical and estimated figures due to the more precise information provided, particularly regarding survival and incidence trends. In conclusion, this work shows the potential of the cancer registries not only to represent their population with respect to cancer morbidity but also as an invaluable tool to extrapolate this information to the larger areas they represent. PMID- 10665858 TI - The prevalence of cancer: a review of the available data. AB - AIMS AND BACKGROUND: Cancer prevalence in a population, defined as the proportion - or the number - of people who were diagnosed with a cancer during their lives and are still alive at a given date, is a crucial indicator for heath care planning and resource allocation. Long-term population-based cancer registries (CR) are the appropriate tools to produce prevalence figures, which, however, are scarcely available. This paper contains a review up to 1999 of the published data world-wide (reports and articles) on cancer prevalence: including measured and estimated figures. MATERIALS AND METHODS: Data on cancer prevalence from CRs are available for the Nordic countries, Connecticut, and Italy. In addition, electronic data are available for the European Union (EU). Data for the Nordic countries were first published in the mid-seventies, reporting the prevalence for 1970. The first data from Connecticut were available 10 years later. Estimates for all EU countries were published by the International Agency for Research on Cancer (IARC) in 1997. In Italy, observed and estimated data on the prevalence of respiratory and digestive tract cancer and breast cancer have been published during the nineties, followed by a systematic analysis for all cancers in 1999. By using information obtained from CRs, cancer prevalence data were calculated directly (observed prevalence) by means of incidence and follow-up information on individual cancer patients, or indirectly (estimated prevalence) by means of mathematical models, which generally use epidemiological information at the aggregate level. RESULTS: Cancer prevalence for all cancers combined (proportions per 100,000 inhabitants) showed values of less than 700 in males and less than 800 in females in 1970 (Finland) to over 2,300 in males and over 3,000 in females in 1992 (Italian registries). With few exceptions, in each country and period considered the cancer sites contributing most to cancer prevalence are lung, colon-rectum, prostate and bladder in males, colon-rectum, breast, uterus (both cervix and corpus) and ovary in females. At present, comparison of measurements from different areas is difficult because there exists no standardized mode of presentation. CONCLUSIONS: In spite of their being potentially useful for health care planning, prevalence data have been produced inconsistently and late by cancer registries, at least in comparison with the systematic availability of incidence and survival statistics. The available data can be compared only to a limited extent due to differences in completeness, in the choice of indicators, in the standard populations, and in the frequency of publication. It would be desirable that in the future data will be produced systematically, with a higher level of standardization compared to the past, and, most importantly, on the same geographic and administrative scale as health-care decision-making. PMID- 10665859 TI - Cancer prevalence. What for? PMID- 10665860 TI - The role of cancer registration for programming health services. AB - The Italian National Health Plan 1998-2000 indicates quantitative and qualitative goals in the fight against cancer. This approach stresses the need of reliable and updated descriptive data to evaluate, at a population level, the burden of neoplastic disease, the results of primary and secondary preventive actions, and the efforts towards a more equal distribution of diagnostic and therapeutic services. The aims of this paper is to evaluate the use of descriptive data to quantify the burden of neoplastic disease, using the data provided by the network of Italian cancer registries (the most reliable source of information on neoplastic disease in Italy). Crude rates are the most adequate for describing the "burden" of cancer patients who are expected in a certain period and will need specific diagnostic or therapeutic activities. Incidence, prevalence and mortality rates provide information on different phenomena (i.e., patients requiring diagnostic and therapeutic activities related with the first definition and treatment of the disease; patients requiring periodic follow-up or treatment of disease relapse; need palliative care). The use of these measures combined is highly informative in relation with the different objectives of health planners (i.e., patients requiring diagnostic and therapeutic activities related with the first definition and treatment of the disease; patients requiring periodic follow up or treatment of disease relapse; need palliative care). PMID- 10665861 TI - MRA in patients with cerebrovascular disease. Considerations of clinical effectiveness. AB - OBJECTIVE: To establish in which clinical contexts cerebral MR angiography (MRA) is routinely carried out in a neurological university department and to describe its clinical impact. MATERIAL AND METHODS: Medical records, reports of findings and documentation of imaging examinations carried out in all 69 patients referred to the Department of Radiology from the Department of Neurology between 1995 and 1998 for cerebral MRA were evaluated. The clinical impact of all imaging findings was assessed on the basis of the medical records. RESULTS: Circulatory disturbances in the vertebrobasilar arteries (n = 34) were the most frequent indication for investigation. MRA followed CT or duplex sonography in 66 of the 69 patients with a mean delay of 8 days. MRA was considered diagnostically inferior to conventional MR in 11 cases, comparable in 30 and superior in 25. Comparing MRA and duplex sonography, the corresponding figures were 12, 29 and 23. In retrospect, 56 MRAs were judged unnecessary. CONCLUSION: Controlled clinical studies on optimal use of MRA are needed to avoid wasting resources and to exploit the method's full diagnostic potential in appropriate cases. PMID- 10665862 TI - Evaluation of dural sinus invasion and extension of extra-axial intracranial tumors. The advantages of a high-resolution postcontrast 3-D gradient-echo technique. AB - OBJECTIVE: To assess the usefulness of a postcontrast 3-D Fourier transform (3DFT) gradient-echo (GRE) technique in dural sinus invasion and extension of extraaxial intracranial tumors in comparison with a conventional spin-echo (SE) technique. MATERIAL AND METHODS: Fourteen consecutive patients with 15 extra axial tumors in contiguity with the dural sinus, including 14 meningiomas and 1 adenoid cystic carcinoma, underwent postcontrast T1-weighted SE and GRE MR studies. Detectability of dural sinus invasion and extension was evaluated using two sequences by two neuroradiologists in a blinded manner and compared with surgical results. Quantitative analysis was also performed to calculate the contrast-to-noise ratio (CNR) between lesion and dural sinus on SE and GRE images. The data were analyzed statistically using a matched paired t-test. RESULTS: In the qualitative evaluation, the detectability of dural sinus invasion in 3DFT-GRE images was superior to that using SE images. The mean CNR for all lesions was 3.86 on SE images and 5.63 on 3DFT-GRE images (p = 0.03). CONCLUSION: For evaluation of dural sinus invasion and the extension of extra-axial tumors, postcontrast 3DFT-GRE MR images were superior to conventional SE images. PMID- 10665864 TI - Does diving damage the brain? MR control study of divers' central nervous system. AB - PURPOSE: To evaluate the prevalence of cerebral white matter changes on MR imaging in healthy elderly compressed air divers with a long diving history in comparison with control subjects who have never dived. MATERIAL AND METHODS: The investigation employed 59 experienced elderly divers and 48 control subjects matched for age, body mass index, alcohol and smoking history. MR studies included a fluid attenuated inversion recovery sequence and T1- and T2-weighted pre- and postcontrast images in axial orientation of the whole brain to localize white matter changes. RESULTS: MR images did not show any morphologic abnormalities in the brains of divers. Both groups - divers and controls - did not differ significantly with respect to white matter changes of the brain. CONCLUSION: No increased prevalence of cerebral white matter changes in compressed air divers compared with a healthy worker sample of similar age were found. Thus, extensive compressed air diving may not necessarily be related to radiological changes on MR. PMID- 10665863 TI - CT of the brain in tuberculous meningitis. A review of 289 patients. AB - PURPOSE: In this retrospective study, CT findings of 289 patients with tuberculous meningitis (TBM) are presented and diagnostic criteria are discussed. MATERIAL AND METHODS: The medical records of patients who were diagnosed as having central nervous system tuberculosis were investigated. Cranial CT investigations of 289 patients with TBM were reviewed. Of these 289 patients, 214 were children and 75 adults; 157 patients were male and 132 were female. CT images were obtained with and without i.v. contrast administration. RESULTS: CT findings were normal in 35 patients and abnormal in 254. The abnormalities were hydrocephalus (172 children, 32 adults), parenchymal enhancement (56 children, 6 adults), contrast enhancement of basal cisterns (32 children, 17 adults), cerebral infarct and focal or diffuse brain edema (29 children, 10 adults), and tuberculoma (9 children, 5 adults). CONCLUSION: CT is pathologic in the great majority of patients with TBM and is helpful in assessing the complications associated with the disease. PMID- 10665865 TI - Abnormalities on brain MR images during the course of familial haemophagocytic lymphohistocytosis in a child. A case report. AB - PURPOSE: To describe and report the neuroradiological findings and clinical features in a patient with familial haemophagocytic lymphohistocytosis (FHL), a rare hereditary immune dysregulation in early childhood characterised by multisystem involvement, including in approximately 30% of cases also the central nervous system (CNS). MATERIAL AND METHODS: Serial brain MR examinations were carried out in a 4.5-year-old boy with FHL, finally complicated with Epstein-Barr virus (EBV)-driven posttransplantation lymphoma. RESULTS: Multiple brain MR examinations before and after contrast enhancement showed discrete perivascular non-enhancing areas of high signal intensity on T2 images, and later also an enhancing lesion in the right caudate nucleus. CONCLUSION: FHL should be included in the differential diagnosis of patchy white matter abnormalities in young patients. EBV-driven post-transplantation lymphoma, which may present as meningial and/or parenchymal CNS infiltration, is a differential diagnostic problem. PMID- 10665866 TI - MR appearance of paraganglioma of the cauda equina. Case reports. AB - PURPOSE: To investigate the value of MR imaging for preoperative diagnosis of paraganglioma of the cauda equina. MATERIAL AND METHODS: A retrospective review of 2 cases of paraganglioma of the cauda equina examined with MR imaging was undertaken. Features assessed included the homogeneity of the lesions, presence or absence of serpiginous flow void and thin hypointense margins. RESULTS: In case 1, the tumor was hyperintense on the postcontrast examination and serpiginous flow void suggested vessels in the upper pole of the tumor. In case 2, the tumor was encapsulated by a thin hypointense margin on both T1- and T2 weighted images, which suggested hemosiderin. CONCLUSION: The MR appearance may be of great value in the preoperative diagnosis of paraganglioma of the cauda equina. PMID- 10665867 TI - Dynamic MR imaging of mandibular osteoradionecrosis. AB - PURPOSE: Osteoradionecrotic bone has been characterised as hypovascular and metabolically inactive tissue with impaired perfusion. The present study was conducted to determine if dynamic contrast-enhanced MR imaging could provide additional information about the vascularity of radionecrotic mandibular bone. MATERIAL AND METHODS: Dynamic contrast-enhanced MR imaging was performed on 10 patients with mandibular osteoradionecrosis (ORN), and on 6 patients, irradiated for oropharyngeal tumours, without symptoms or signs of ORN. Nine patients in the ORN group received a series of 20 hyperbaric oxygen (HBO) treatments, after which the dynamic MR investigation was repeated. RESULTS AND CONCLUSION: Radiation per se did not lead to increased contrast enhancement, whereas all patients with ORN showed marked contrast enhancement of the osteoradionecrotic bone marrow. After HBO treatment, pathological contrast enhancement of the abnormal bone marrow could still be seen, but the rate of enhancement was less than before in 7 of 9 patients. Two patients had an increase in the enhancement rate. The findings suggest the existence of an increased and patent microvasculature. PMID- 10665868 TI - Early glottic squamous cell carcinoma. Predictive value of MR imaging for the rate of 5-year local control with radiation therapy. AB - PURPOSE: To evaluate MR findings in early (T1 and T2 stages) glottic carcinomas and the predictive value of MR imaging for the rate of 5-year local control with radiation therapy. MATERIAL AND METHODS: Eighty-three patients with early glottic carcinomas were prospectively examined with MR at 1.5 T. MR investigation included unenhanced T1-weighted, T2-weighted, dynamic and contrast-enhanced T1 weighted images. Three patients with presumed advanced diseases on MR were initially treated with total laryngectomy and were excluded from the study. The remaining 80 patients were treated with radiation therapy with curative intent. Tumor detectability, size and relationship to the thyroid cartilage were determined on MR images. The MR findings were then correlated with the rate of local control. RESULTS: Forty-eight of 80 lesions (60%) were detected on MR imaging. All detected lesions but 1 demonstrated increased signal on T2-weighted images. The lesions were best delineated on dynamic images (statistically significant). The 5-year local control rate with radiation therapy was 72%. Univariate analysis revealed clinical T stage, MR detectability, tumor size and relationship to the thyroid cartilage as significant predictors. Multivariate analysis revealed that the relationship to the thyroid cartilage was an independent factor. CONCLUSION: MR provides prognostic information about the results of definitive radiation therapy. To evaluate the tumor extension in lesions detected on precontrast MR images, contrast-enhanced dynamic images should be obtained. PMID- 10665869 TI - Doppler ultrasonography in predicting relapse of hyperthyroidism in Graves' disease. AB - PURPOSE: To determine whether Doppler ultrasonography could be useful in the prediction of relapse of hyperthyroidism in patients with Graves' disease. MATERIAL AND METHODS: Forty patients with Graves' disease confirmed by laboratory tests were examined for a number of blood flow parameters in the inferior thyroid artery before and after they were subjected to proper antithyroid drug treatment. Data were retrospectively reviewed and compared with findings for a control group of 16 age-matched subjects. RESULTS: Significantly increased blood flow parameters were observed both in patients with active hyperthyroidism before treatment and in euthyroid patients who presented a relapse shortly after withdrawal of proper antithyroid drug treatment versus normal controls. Conversely, no significant differences were observed between patients who remained in stable remission and normal controls. CONCLUSION: Our results support the concept that Doppler ultrasonography evaluation of patients with Graves' disease may contribute to the detection of a relapsing course of hyperthyroidism. PMID- 10665870 TI - CT findings in tuberculous otomastoiditis. A case report. AB - PURPOSE: Otomastoiditis is a rare but important manifestation of tuberculosis and is well recognizable when information on its clinical course is considered in connection with the radiographic changes. MATERIAL AND METHOD: A patient with a clinical history of chronic otorrhea, resistant to conventional therapy but without dramatic symptoms, was referred for CT examination. RESULTS: CT revealed widespread soft tissue densities in the tympanic cavity and in the mastoid process, with bone erosions in the latter. Surgery and bacteriology confirmed the diagnosis of Mycobacterium tuberculosis infection. CONCLUSION: CT evidence of widespread bone destruction without clinical signs of aggressive infection should suggest the diagnosis of a mycobacterial process. Early treatment is essential in order to avoid propagation of the disease and lasting loss of function. PMID- 10665871 TI - Effect on sensitivity and specificity of mammography screening with or without comparison of old mammograms. AB - PURPOSE: To evaluate the effect of old mammograms on the specificity and sensitivity of radiologists in mammography screening. MATERIAL AND METHODS: One hundred and fifty sets of screening mammograms were examined by 3 experienced screeners twice: once without and once in comparison with older mammograms. The films came from a population-based screening done during the first half of 1994 and comprised all 35 cancers detected during screening in 1994, 12/24 interval cancers, 14/34 cancers detected in the following screening and 89 normal mammograms. RESULTS: Without old mammograms, the screeners detected an average of 40.3 cancers (range 37-42), with a specificity of 87% (85-88%). With old mammograms, the screeners detected 37.7 cancers (range 34-42) with a specificity of 96% (94-99%). The change in detection rate was not significant. However, the increase in specificity was significant for each screener (p = 0.0002-0.03). CONCLUSION: Mammography screening with old mammograms available for comparison decreased the false-positive recall rate. The effect on sensitivity, however, was unclear. PMID- 10665872 TI - Pre-operative simultaneous stereotactic core biopsy and fine-needle aspiration biopsy in the diagnosis of invasive lobular breast carcinoma. AB - PURPOSE: To determine the diagnostic value of stereotactic core needle biopsy (SCNB) in comparison to stereotactic fine-needle aspiration biopsy (SFNAB) in patients with invasive lobular carcinoma (ILC). MATERIAL AND METHODS: Twenty-two patients with clinical or mammographic findings suspicious of malignancy underwent surgery where postoperative histopathology showed ILC. Pre-operative attempts of diagnosis were made using SFNAB and SCNB. SFNAB was done with a spinal needle 0.7- or 0.9-mm and SCNB was simultaneously performed with an automated 2.1-mm biopsy gun in all patients. RESULTS: SFNAB was diagnostic of carcinoma in 9 women, showed "probable carcinoma" in 5 and "atypia" in 3. In the remaining 5 women, SFNAB showed no atypia. SCNB diagnosed ILC in 20 patients and showed ILC as well as invasive ductal carcinoma (IDC) in 1. Ductal carcinoma in situ was suggested in the remaining patient. CONCLUSION: SCNB was superior to SFNAB in diagnosing ILC and did not miss any carcinoma, whereas SFNAB was non diagnostic in 8 cases. SCNB is thus recommended in patients with suspicion of ILC of the breast. PMID- 10665873 TI - Primary tuberculosis of the breast. A case report. AB - The mammographic and US features of a case of primary tuberculosis of the breast are presented. The differential diagnosis with other benign or malignant conditions of the breast can be difficult with imaging methods due to the variable pattern of presentation of such an inflammatory lesion. PMID- 10665874 TI - High-resolution imaging of coronary calcifications by intense low-energy fluoroscopic X-ray obtained from synchrotron radiation. AB - PURPOSE: To obtain an intense monochromatic low-energy X-ray from synchrotron radiation (SR) and apply it to detect coronary calcifications. METHODS AND RESULTS: The SR beam was reflected with a silicon crystal to be expanded (150 mm in height and 80 mm in width) and to be monochromatized at an energy level of 37 keV. The X-ray was intermittently irradiated to obtain dynamic imaging of 30 images/s. Images were recorded by a digital fluorography system. The low-energy X ray from SR sharply visualized calcification of coronary arteries, while conventional X-ray could not visualize coronary calcification. CONCLUSION: The intense monochromatic low-energy X-ray from SR is sensitive, has high-resolution for imaging coronary calcification and may serve as a screening method for coronary artery disease. PMID- 10665875 TI - A double coaxial delivery system for deployment of a two-stage stent-graft. An experimental feasibility study. AB - PURPOSE: To design and test a delivery system for successive, rapid two-stage deployment of an aortic stent graft. MATERIAL AND METHODS: An aortic stent graft was made that consisted of two separate parts. A delivery system composed of two independent coaxial mechanisms was fabricated and used to deploy the stages of the graft in dogs' aortas. RESULTS: Delivery was successful in all four dogs, and required less than 2 min in each animal. CONCLUSION: The double coaxial delivery system enabled quick endovascular assembly of a two-stage aortic stent-graft. PMID- 10665876 TI - Infrapopliteal percutaneous transluminal angioplasty for limb salvage. AB - PURPOSE: To evaluate long-term results of infrapopliteal percutaneous transluminal angioplasty (PTA) for limb salvage. MATERIAL AND METHODS: A retrospective study of 71 consecutive infrapopliteal PTAs in 49 patients with rest pain (n = 20) or ulceration (n = 29) was conducted. In 18 patients, surgical minor amputation or debridment was also performed. RESULTS: Technical success was achieved in 45 patients. Four failures necessitated 2 amputations. One patient died in the postoperative course. Global morbidity rate was 16%, including minor complications in 5 patients and major vascular complications in 3 patients. After technical success during the follow-up (median duration 21 months), restenoses occurred in 4 patients, of whom 3 had a successful re-PTA (clinical success rate 72%). Survival, primary patency, secondary patency and limb salvage rates were, respectively, 75%, 81%, 88% and 87% after 3 years. The only positive predictive factor for primary patency was the presence of diabetes mellitus. CONCLUSION: Infrapopliteal PTA is a safe and effective procedure, allowing good patency and limb salvage rates with low mortality and morbidity. PMID- 10665877 TI - High-resolution MR imaging of the carpal tunnel and the wrist. Application of a 5 cm surface coil. AB - PURPOSE: To make a comparative analysis of transversal tomograms obtained by high resolution MR imaging with frozen cross-sections of an anatomical forearm specimen. Twenty-two healthy volunteers were also examined using the same coil system to test for a range of possible clinical applications and for the depiction of morphological and morphometrical values of normal anatomy in vivo. MATERIAL AND METHODS: MR images of the carpal tunnel of 22 healthy volunteers were obtained with a 1.5-T whole-body system with a 5-cm surface coil. Measurements were recorded with a field-of-view between 50x50 mm2 and 60x60 mm2 in a 256x256 pixel matrix for the T1 sequence. A slice thickness of 2 mm was used. The images were acquired using a T1-weighted SE sequence (TR/TE 500/38 ms) and a T2-weighted SE sequence (TR/TE 2000/70 ms). Additionally, a formalin-fixed anatomical forearm specimen was imaged for anatomic correlation. The imaged transversal cross-section levels in the specimen were subsequently freeze sectioned. The anatomical structures of the MR findings were identified and compared with the macroscopical sections of the specimen. RESULTS: Based on the good depiction of details at this coil system with a pixel size in T1 of 0.195x0.195 mm, high-resolution MR imaging enabled identification of the interior structures of the carpal tunnel, as well as delineation of connective tissue. The clinical value of high-resolution MR includes the diagnosis of carpal tunnel syndrome and inflammatory disorders of the wrist. CONCLUSION: Our results support the feasibility of high-resolution MR imaging of the carpal tunnel and the wrist using small surface coils. PMID- 10665878 TI - High-resolution MR imaging of the knee at 3 T. AB - PURPOSE: To examine the practical feasibility of using a 3.0-T MR unit to obtain high-quality, high-resolution images of the knee joint. MATERIAL AND METHODS: One human cadaveric and 5 porcine knees were imaged with the 3.0-T unit. Sets of T1 weighted spin echo images were obtained with in-plane resolution of 0.195x0.39 mm and an acquisition time of approximately 5 min. Two porcine knees were also imaged with the 1.0-T unit with an identical imaging protocol and the signal-to noise (S/N) ratios were measured on images at 3 T and 1 T. RESULTS: The 3-T MR system provided detailed delineation of the knees. Deep layers of the medial collateral ligament and associated fine fibers beneath the medial and lateral collateral ligament were demarcated. We observed precise demonstration of the tibial attachment of the anterior cruciate ligament, irregularity of the meniscal free edge, and conjoint tendon formation together with the lateral collateral ligament and the biceps femoris tendon. Compared to the 1-T unit, the S/N ratio with the 3-T unit was increased by a factor of 1.39 to 1.72. CONCLUSION: Due to the potential advantage of obtaining detailed images, the 3-T MR system suggests a practical utility for fine demonstration of the knee morphology. PMID- 10665879 TI - MR findings of desmoplastic fibroma of the spine. A case report. AB - We report on the MR imaging findings in a case of spinal desmoplastic fibroma (DF). DF of the bone is a rare, locally aggressive tumor. It is commonly located in long bones, pelvis or mandible. DF involving the spine is extremely rare and difficult to distinguish from other bony lesions such as giant cell tumor, chordoma and fibrous dysplasia of the spine. This case of DF of the spine showed MR findings similar to those of DF arising in the metaphysis of a long bone. PMID- 10665880 TI - Glycogen content in rat liver. Importance for CT and MR imaging. AB - PURPOSE: CT and MR imaging are appropriate modalities for imaging of the liver. Contrast media are used to obtain a greater difference in attenuation and signal intensity, respectively, between normal liver tissue and focal lesions. However, no studies have attempted to determine whether physiological nutritional status of the liver during fasting is of importance for the native signal of normal liver tissue. MATERIAL AND METHODS: Using normal and fasting rats, we performed hepatic CT and MR imaging and glycogen analyses from excised tissue. RESULTS: A significantly higher liver attenuation in normal rats compared to fasting rats was found in CT. In MR images, there was a small but significantly lower liver signal-to-noise ratio in normal rats compared to fasting rats in T1-weighted and proton density-weighted images. Glycogen analyses showed depleted glycogen deposits in fasting rats and a mean glycogen content of 50.1 mg glucose equivalent/g liver tissue in normal rats. CONCLUSION: In CT, a normal nutritional status increases the native attenuation in normal liver tissue. The changes in attenuation in normal liver tissue correlate well with the additional attenuation of glycogen storage in the hepatocyte. The results indicate that the nutritional status is of less importance in MR imaging. PMID- 10665881 TI - Impact of intraoperative ultrasonography on surgical treatment of liver tumours. AB - PURPOSE: The aim of this study was to evaluate the impact of intraoperative ultrasonography (IOUS) on surgical decision-making in patients with liver tumours. MATERIAL AND METHODS: IOUS of the liver was performed in 116 patients undergoing surgery for liver tumours. The patients were evaluated preoperatively by ultrasonography, CT and in some cases, by ERCP and scintigraphy. IOUS findings were compared with the results of preoperative imaging and surgical findings. RESULTS: The surgical procedures were altered in 50 cases (43%), extended in 19 cases (16%) and reduced in 8 cases (7%). Twenty-three patients (20%) were found inoperable. Intraoperative evaluation caused surgical modification by IOUS in 13 patients (11%), by surgical findings in 14 patients (12%) and by a combination of both in 23 patients (20%). CONCLUSION: IOUS is a precise diagnostic method for staging the operability of liver tumours. Unnecessary surgical procedures can be avoided. In total, IOUS altered the preoperative plan in one-third of our patients and is therefore recommended as a routine procedure in connection with major liver surgery. PMID- 10665882 TI - Structure and content of 400 CT reports in four teaching hospitals using a new, Windows-based software tool. AB - PURPOSE: To test a new software tool developed for analysis of radiology reports and to compare CT reports from four different teaching hospitals. MATERIAL AND METHODS: Four hundred CT reports were randomly collected from four Finnish university hospitals. A Windows 3.1-based software tool was developed to make a comparative analysis of the information content of CT reports. The structure of the reports was partly analyzed manually. RESULTS: The new software tool greatly facilitated semiquantitative analysis of the information content of residents' and radiologists' reports. There were some local differences in the length and structure of the reports, the choice of vocabulary and the number of differential diagnoses given, and large differences in the use of an "impression" section. Thorough description of focal lesions was included in less than 50% of the reports from each of the four institutions. CONCLUSION: The variation in well structured CT reports indicates considerable geographic differences in radiology reporting, which may reflect the long-term influences of a few teachers. Rational application of the communication standards should improve the quality of written radiology reports. Such standards should be emphasized, particularly in teaching hospitals. PMID- 10665883 TI - Analysis of the availability and completeness of previous radiological examinations related to plain films. AB - PURPOSE: To evaluate the availability and completeness of previous radiological examinations. MATERIAL AND METHODS: Seven different types of examinations were analysed separately: 1) chest examinations, 2) bedside chest examinations, 3) skeletal examinations, 4) angiographic examinations, 5) mammographic examinations, 6) CT examinations, and 7) MR examinations. A retrospective part of the study covered the calendar year 1997, while a prospective part referred to 1998. The sample size in each modality and each part of the study was 100 observations, resulting in a total number of 1,400 examinations. RESULTS: The average availability and completeness of radiographs in the retrospective analysis was 96.9% in 1997. The average availability and completeness of radiographs in the prospective analysis was 85.5% for the year 1998. This applied to necessary previous examinations of the same or different modality irrespective whether the examination was conducted internally or externally. CONCLUSION: The real loss rate of radiographs derived from the retrospective investigation was 3.1%. The prospective part of the study showed that 14.5% of the required previous internal and external examinations of the same or different modality were not available completely and in time to the examining radiologist. PMID- 10665884 TI - The autopsy and the living. Autopsy Committee of the College of American Pathologists. PMID- 10665885 TI - Melatonin therapy: from benzodiazepine-dependent insomnia to authenticity and autonomy. PMID- 10665887 TI - Billions for defense: the pervasive nature of defensive medicine. PMID- 10665886 TI - The lost art of auscultation. PMID- 10665888 TI - An update on perioperative management of diabetes. AB - Surgery in the patient with diabetes mellitus is relatively common, as the numbers of persons with diabetes is increasing and diabetes predisposes to medical conditions that require surgical intervention. An estimated 25% of diabetic patients will require surgery, and advances in perioperative care of these patients allow them to safely undergo the most complicated surgical procedures. We will review issues of preoperative, intraoperative, and postoperative care of diabetic patients. PMID- 10665889 TI - Cigar smoking and death from coronary heart disease in a prospective study of US men. AB - CONTEXT: The prevalence of cigar smoking has increased rapidly in the United States since 1993. Although cigarette smoking is known to be an important cause of coronary heart disease (CHD) mortality, the relationship between cigar smoking and CHD mortality is unclear. OBJECTIVE: To determine whether cigar smoking increases risk of CHD mortality. DESIGN: Prospective cohort study with follow-up for mortality from 1982 through 1991. SETTING: United States. PARTICIPANTS: A total of 121 278 men, aged 30 years and older, in the American Cancer Society's nationwide Cancer Prevention Study II cohort who completed a baseline questionnaire on smoking history and other risk factors in 1982, had never smoked cigarettes or pipes, and had no diagnosed heart disease or diabetes at baseline. MAIN OUTCOME MEASURE: Death from CHD recorded as the underlying cause of death on the death certificate. RESULTS: There were 2508 deaths from CHD from 1982 through 1991. The association between cigar smoking and death from CHD was stronger among younger men and current rather than former smokers, as is observed with cigarette smoking. No increased risk was observed among current cigar smokers aged 75 years or older, or for former cigar smokers of any age. For men younger than 75 years who were current cigar smokers at baseline, the adjusted rate ratio for CHD mortality was 1.30 (95% confidence interval, 1.05-1.62). CONCLUSIONS: These results suggest that smoking cigars increases risk of early death from CHD. Any adverse effect of cigars on CHD is of particular importance given the rapidly rising prevalence of cigar smoking in the United States. PMID- 10665890 TI - Asthma self-management: do patient education programs always have an impact? AB - BACKGROUND: During the past 15 years, programs to improve self-management practices in adults with asthma have reported improvement in functional status and reduction of inappropriate use of health care services. However, these programs usually represent an ideal approach, applying multiple patient education methods. Consequently, when these programs are found to be efficacious, it is important to replicate the programs as well as to evaluate less complex methods that may be more appropriate for nonacademic health care settings. METHODS: We compared the following 3 standardized self-management treatments in a randomized, controlled trial: (1) a replication of the self-management program developed at a university medical center that was previously shown to be efficacious; (2) a modified version of this program including only the core elements; and (3) a usual-care program. Outcome measures included medication and inhaler regimen adherence, asthma symptoms, respiratory illness, functional status, and use of health care resources. RESULTS: All 3 groups improved on measures of respiratory illnesses, use of health care services, and functional status. Patients in both education groups did no better than the usual-care group. CONCLUSIONS: The results are inconsistent with the results of the first asthma self-management study at this institution and with those of efficacy studies of similar programs. Two factors, selection of the patient population and historical changes in asthma treatment, most likely contributed to the lack of impact of the self-management programs. As a result of the improved standards for usual care due to both factors, the opportunity to effect patient outcomes was substantially reduced. PMID- 10665891 TI - Persistent stress as a predictor of genital herpes recurrence. AB - BACKGROUND: Results of several studies suggest that psychological stress and negative mood can trigger genital herpes recurrences, but results are inconsistent. OBJECTIVE: To determine whether short-term or persistent psychological stress or specific negative moods are predictive of genital herpes recurrences in women. METHODS: A prospective cohort study followed up participants for 6 months using weekly assessments of stress and mood, monthly assessments of life change events, and diary reports of genital herpes recurrences confirmed by medical examination when feasible. The community sample consisted of 58 women, aged 20 to 44 years, with a 1- to 10-year history of visible genital herpes recurrence and at least 1 recurrence in the previous 6 months. RESULTS: Persistent stress predicted recurrence in the subsequent week (odds ratio, 1.08 per unit increase in stress; 95% confidence interval, 1.01 1.15; P=.03). After adjusting for recurrence in the previous week, the more weekly persistent stress reported, the greater the likelihood of recurrence the following week. Also, an increased recurrence rate occurred after the month during which participants experienced their highest levels of anxiety (P =.03). There were no significant associations between recurrence and short-term stress, life events, depressive mood, anger, or phase of menstrual cycle. CONCLUSIONS: Persistent stressors and highest level of anxiety predicted genital herpes recurrence, whereas transient mood states, short-term stressors, and life change events did not. Women with herpes can be reassured that short-term stressful life experiences and dysphoric mood states do not put them at risk for increased outbreaks of recurrent genital herpes. PMID- 10665892 TI - The health and economic benefits associated with pneumococcal vaccination of elderly persons with chronic lung disease. AB - BACKGROUND: More than 50% of the elderly population has not received pneumococcal vaccination. Uncertainty regarding the benefits of immunization, particularly for noninvasive disease, may contribute to the underuse of pneumococcal vaccine. OBJECTIVE: To assess the health and economic benefits associated with pneumococcal vaccination. METHODS: We conducted a 2-year retrospective cohort study among all elderly members of a staff-model managed care organization who had a baseline diagnosis of chronic lung disease. The study outcomes were assessed over 2 years, from November 15, 1993, through November 14, 1995, and included hospitalizations for pneumonia and influenza, death, and hospitalization costs. Using administrative data, we compared these outcomes for vaccinated and unvaccinated subjects using multivariate models to control for subjects' baseline demographic and health characteristics. The additive benefits of combined influenza and pneumococcal vaccination were also assessed for the 2 influenza seasons included in the study. RESULTS: There were 1898 subjects. Pneumococcal vaccination was associated with significantly lower risks for pneumonia hospitalizations (adjusted risk ratio [RR], 0.57; 95% confidence interval [CI], 0.38-0.84; P=.005) and for death (adjusted RR, 0.71; 95% CI, 0.56-0.91; P = .008). For the control outcome of all nonpneumonia hospitalizations, rates did not differ significantly between the 2 groups (adjusted RR, 0.91; 95% CI, 0.77 1.07; P= .24). During the influenza seasons included in the study, the benefits of pneumococcal and influenza vaccinations were additive, with an adjusted RR of 0.28 (95% CI, 0.14-0.58; P<.001) for the number of hospitalizations for pneumonia and influenza among persons who had received both vaccinations compared with those who had received neither and an adjusted odds ratio of 0.18 (95% CI, 0.11 0.31; P<.001) for death. Over the 2-year outcome period, pneumococcal vaccination was also associated with direct medical care cost savings. CONCLUSIONS: Pneumococcal vaccination of elderly persons with chronic lung disease was associated with fewer hospitalizations for pneumonia, fewer deaths, and direct medical care cost savings. PMID- 10665893 TI - Early switch from intravenous to oral antibiotics and early hospital discharge: a prospective observational study of 200 consecutive patients with community acquired pneumonia. AB - To determine the proportion of patients who can be treated with early switch to oral antibiotics and early discharge, to evaluate clinical outcome and patient satisfaction for patients treated with early switch and early discharge, and to define the factors that interfere with early discharge for some of the patients who underwent early switch to oral antibiotic therapy. DESIGN: Prospective study. PARTICIPANTS: Two hundred consecutive hospitalized patients with community acquired pneumonia. MAIN OUTCOME MEASURES: Number of days needed to switch to oral therapy and length of hospital stay. Clinical outcome and satisfaction with care were evaluated for those patients treated with early switch and early discharge. RESULTS: Early switch to oral antibiotics (within the first 3 days of hospitalization) was performed in 133 patients (67%). Clinical failure was documented in 1 patient. Early switch and early discharge was performed in 88 patients (44%). The mean length of hospital stay for this group was 3.4 days. The most common reason for prolonged hospitalization after the switch to oral antibiotics was the need for diagnostic workup. More than 95% of patients were satisfied with the care they had received. CONCLUSIONS: Using simple clinical and laboratory criteria, a significant proportion of hospitalized patients with community-acquired pneumonia (44%) can be treated with early switch and early discharge. This model did not affect patient outcome, decreased the length of hospitalization, and was associated with a high level of patient satisfaction. PMID- 10665894 TI - Facilitation of benzodiazepine discontinuation by melatonin: a new clinical approach. AB - BACKGROUND: Benzodiazepines are the most frequently used drug for the treatment of insomnia. Prolonged use of benzodiazepine therapy is not recommended. However, many patients, particularly older patients, have difficulties discontinuing therapy. Melatonin, a hormone that is produced at night by the pineal gland, promotes normal sleep in humans and augments sleep induction by benzodiazepine therapy. OBJECTIVE: To assess whether the administration of melatonin could facilitate the discontinuation of benzodiazepine therapy in patients with insomnia. METHODS: Thirty-four subjects receiving benzodiazepine therapy were enrolled in the 2-period study. In period 1, patients received (double-blinded) melatonin (2 mg in a controlled-release formulation) or a placebo nightly for 6 weeks. They were encouraged to reduce their benzodiazepine dosage 50% during week 2, 75% during weeks 3 and 4, and to discontinue benzodiazepine therapy completely during weeks 5 and 6. In period 2, melatonin was administered (single-blinded) for 6 weeks to all subjects and attempts to discontinue benzodiazepine therapy were resumed. Benzodiazepine consumption and subjective sleep-quality scores were reported daily by all patients. All subjects were then allowed to continue melatonin therapy and follow-up reassessments were performed 6 months later. RESULTS: By the end of period 1, 14 of 18 subjects who had received melatonin therapy, but only 4 of 16 in the placebo group, discontinued benzodiazepine therapy (P = .006). Sleep-quality scores were significantly higher in the melatonin therapy group (P = .04). Six additional subjects in the placebo group discontinued benzodiazepine therapy when given melatonin in period 2. The 6-month follow-up assessments revealed that of the 24 patients who discontinued benzodiazepine and received melatonin therapy, 19 maintained good sleep quality. CONCLUSION: Controlled-release melatonin may effectively facilitate discontinuation of benzodiazepine therapy while maintaining good sleep quality. PMID- 10665895 TI - The association between antecedent vancomycin treatment and hospital-acquired vancomycin-resistant enterococci: a meta-analysis. AB - BACKGROUND: The association between vancomycin hydrochloride treatment and vancomycin-resistant enterococci (VRE) has been investigated in numerous studies with variable results. OBJECTIVES: To conduct a meta-analysis to estimate the magnitude of the association between vancomycin treatment and individual risk of VRE and to identify study characteristics that accounted for heterogeneity in study results. METHODS: Studies were identified using MEDLINE with index terms "Enterococcus," "Enterococcus faecalis," or "Enterococcus faecium" and "vancomycin," "drug resistance," "drug resistance, microbial," or "drug resistance, multiple or risk factors." Reports from conferences and reference lists of recent reviews were used. A total of 420 published reports and 98 conference reports were reviewed; 20 studies described in 15 published reports were included in the analysis. We recorded study period, hospital setting, case and control definitions, length of hospital stay, method of adjustment for differences in length of stay, and data on treatment with vancomycin. The odds ratio (OR) of vancomycin treatment provided the measure of association analyzed. A random-effects model was used to estimate the pooled OR. RESULTS: When results from all 20 studies were combined, the pooled OR was 4.5 (95% confidence interval, 3.0-6.9), but the test for heterogeneity was highly significant (P<.001). The 5 studies that used patients with vancomycin-susceptible enterococci as controls found a stronger association (pooled OR, 10.7; 95% confidence interval, 4.8-23.8) than the 15 studies that used controls who had no VRE isolated (pooled OR, 2.7; 95% confidence interval, 2.0-3.8). After restricting the analysis to the latter studies only, no heterogeneity was evident in the unadjusted study results. Patients with VRE had stayed in the hospital much longer than control patients. Studies that adjusted for this difference found only a small and nonsignificant association between vancomycin treatment and VRE (pooled OR, 1.4; 95% confidence interval, 0.74-2.60). We also detected publication bias, favoring report of studies that found a large measure of association. CONCLUSIONS: The reported strong association between vancomycin treatment and hospital-acquired VRE results from the selection of the reference group, confounding by duration of hospitalization, and publication bias. Studies that accounted for these factors found only a small and nonsignificant association. PMID- 10665896 TI - Diabetes mellitus and nontraumatic lower extremity amputation in black and white Americans: the National Health and Nutrition Examination Survey Epidemiologic Follow-up Study, 1971-1992. AB - BACKGROUND: The comparative long-term risk of non-traumatic lower extremity amputation (LEA) in black and white Americans, 2 groups with strikingly different rates of diabetes mellitus, is not known. OBJECTIVE: To examine the 20-year incidence of LEA in relation to race and diabetes mellitus. METHODS: The 14 407 subjects in the National Health and Nutrition Examination Survey Epidemiologic Follow-up Study were observed prospectively between 1971 and 1992. Prevalent diabetes mellitus was ascertained at the baseline examination, and incident diabetes mellitus, during follow-up. Lower extremity amputation was ascertained from hospital discharge records. Cox regression analysis was used to estimate associations between race, diabetes mellitus, and risk of first LEA. RESULTS: During the study period, 158 LEAs occurred among 108 subjects. While black subjects constituted 15.2% of the cohort, they represented 27.8% of the subjects with amputation (P = .002). The 20-year age-adjusted rate ratio of first LEAs for black subjects-white subjects was 2.14. Regression analyses confirmed the importance of diabetes mellitus as a key LEA risk factor. The association between prevalent diabetes mellitus and LEA risk was substantially higher (relative risk [RR], 7.19; 95% confidence interval [CI], 4.61-11.22) than that for incident diabetes mellitus (RR, 3.15 [CI, 1.84-5.37]), highlighting the importance of diabetes mellitus duration on LEA risk. While preliminary analyses adjusted for age and diabetes indicated a significant association between race and LEA risk (RR, 1.93 [95% CI, 1.26-2.96]), the effect of race diminished (RR, 1.49 [95% CI, 0.95-2.34]) following adjustment for education, hypertension, and smoking. CONCLUSIONS: Although black subjects experienced higher age- and diabetes mellitus-adjusted rates of amputation than their white counterparts, a combination of social and environmental factors may account for the apparent ethnic difference. More research into nonbiological factors associated with LEA may reduce the occurrence of these procedures in both black and white individuals. PMID- 10665897 TI - Cardiopulmonary auscultation: duo for strings--Opus 99. AB - In spite of increasing mechanization in medicine and reliance on "high-tech" diagnostic tools, bedside clinical skills of the attending physician can still identify findings that are missed by the more sophisticated devices. Using a stethoscope, we relied on our skills in inspection, palpation, percussion, auscultation, as well as echocardiography and phonocardiography to diagnose a patient whose murmur was very reminiscent of the D-sharp pizzicato in the Cello Sonata in F, Opus 99, by Johannes Brahms. Initial echocardiography was not helpful. We suspected an anomalous chorda and confirmed this with phonocardiography and a second echocardiography. Although advances in cardiac imaging are extremely helpful, the use of simple clinical skills, in addition to being fun, is not obsolete. Cardiopulmonary auscultation should receive more emphasis in the medical school curriculum and clinical training. PMID- 10665898 TI - Fibromyalgia, chronic fatigue syndrome, and Addison disease. PMID- 10665899 TI - Patients with fibromyalgia must be treated fairly. PMID- 10665900 TI - Helicobacter pylori and gastric cancer: both primary and secondary preventive measures are required. PMID- 10665901 TI - Neurocardiogenic syncope and cancer: a paraneoplastic association? PMID- 10665902 TI - Is garlic an effective treatment for Helicobacter pylori infection? PMID- 10665903 TI - Latex agglutination is a valid method for the measurement of D-dimer levels. PMID- 10665904 TI - To E or not to E. PMID- 10665905 TI - A futurist meets the 21st century: love at first sight. PMID- 10665906 TI - Cardiac autonomic denervation in congestive heart failure: comparison of Chagas' heart disease with other dilated cardiomyopathy. AB - Congestive heart failure (CHF) is associated with activation of the cardiac sympathetic nerves. However, impairment of the sympathetic nerve terminals in patients with CHF has been indicated by studies showing reduction of cardiac norepinephrine uptake and stores. This investigation studies the histochemical evaluation of the sympathetic nerve terminals in CHF. The cardiac parasympathetic innervation was also studied to address the question of specificity of the presumed sympathetic denervation. Nineteen patients with CHF underwent cardiac transplantation or partial ventriculectomy, which provided the heart tissue. In 11 of them, the dilated cardiomyopathy was associated with Chagas' disease. Inflammatory process and fibrosis were studied histologically. The sympathetic and parasympathetic nerve fibers were visualized through histochemical techniques for, respectively, catecholamines and acetylcholinesterase activity. By using a computer-assisted morphometric program, the inflammation, fibrosis, and parasympathetic innervation were quantified. Moderate to severe fibrosing myocarditis characterized the hearts of the chagasic patients. In cardiomyopathies not associated with Chagas' disease, the inflammation was discrete, if present, but the amount of fibrosis was similar to that found in Chagas' cardiomyopathy. Reduction of both kinds of nerve terminals occurred in the heart of all patients. The parasympathetic denervation was proven to be more severe in chagasic cardiomyopathy. Our data on the heart innervation indicate a progressive autonomic denervation in heart failure. In Chagas' heart disease, the denervation seems to be more severe or rapid, probably because of the sustained inflammatory process. PMID- 10665907 TI - GLUT1: a newly discovered immunohistochemical marker for juvenile hemangiomas. AB - Juvenile hemangiomas are common, benign vascular tumors of infancy. These lesions enlarge rapidly through cellular hyperplasia during the first year of life and then involute over several years. Distinctive histopathologic features of hemangiomas diminish during this evolution, and differentiation from vascular malformations becomes increasingly difficult. This distinction has important therapeutic implications, as juvenile hemangiomas differ from malformations in natural history and in potential for recurrence. We report here that high endothelial immunoreactivity for the erythrocyte-type glucose transporter protein GLUT1 is a specific feature of juvenile hemangiomas during all phases of these lesions. In a retrospective study, we found intense endothelial GLUT1 immunoreactivity, involving more than 50% of lesional microvessels, in 97% (139 of 143) of juvenile hemangiomas from patients aged 1 month to 11 years. No endothelial GLUT1 immunoreactivity was found in any of 66 vascular malformations (17 arteriovenous, 33 venous, 11 lymphatic, and 5 port-wine) from patients aged 5 days to 75 years, or in any of 20 pyogenic granulomas or 7 granulation tissue specimens. Abundant Ki-67 positivity in these latter lesions established that GLUT1 expression does not simply reflect mitotically active endothelium. Focal GLUT1 immunoreactivity was found in 3 of 12 angiosarcomas, but not in any of 5 hemangioendotheliomas (epithelioid or infantile kaposiform). These findings establish GLUT1 immunoreactivity as a highly selective and diagnostically useful marker for juvenile hemangiomas. Because high levels of endothelial GLUT1 expression in normal tissue are restricted to microvessels with blood-tissue barrier function, these findings also have implications for the molecular and developmental pathogenic mechanisms of juvenile hemangiomas. PMID- 10665908 TI - Cytological recognition of invasive squamous cancer of the uterine cervix: comparison of conventional light-microscopical screening and neural network-based screening. AB - Cytologic recognition of invasive or microinvasive cancer of the uterine cervix may present substantial difficulties. In this study, we compared conventional light-microscopical screening of 109,104 cervical smears and neural network-based screening (NNS) of 245,527 smears, all obtained by the spatula-Cytobrush method. Two populations of Dutch women were included in the study: those receiving smears within the framework of the Dutch population screening program ("routine smears") and those receiving smears for other reasons, discussed in the text ("interval smears"). There were 71 smears, from an equal number of biopsy-confirmed invasive squamous carcinomas, 28 of which were microinvasive. The "interval smears" yielded a statistical valid higher prevalence of invasive cancer than "routine smears." Except for 5 smears that contained no evidence of abnormality ("sampling errors"), no false-negative errors occurred in the 52 NNS cases, whereas 4 such errors occurred in the 19 conventionally screened cases. By measuring the amount of cancerous material present in each smear (mapping), it could be documented that NNS was effective even in smears with a small number of cancer cells, whereas the 4 conventional false-negative screening errors occurred in smears of this type. The study showed that cells derived from invasive cancer of the cervix may have large bland nuclei that do not fit the images commonly associated with squamous cancer cells. Neural network-based screening of cervical smears was more effective than conventional screening in the diagnosis of invasive squamous cancer of the uterine cervix. PMID- 10665909 TI - Malignant epithelioid vascular tumors of the pleura: report of a series and literature review. AB - Primary malignant vascular tumors of the pleura are rare. The significance and difficulty of distinction between pleural epithelioid hemangioendothelioma (EHE) and angiosarcoma have not yet been addressed. A new series of pleural angiosarcoma is reported, and the relevant literature is reviewed. Five cases were identified from files of the authors' institutions and personal consultation cases (J.J.B.). Twenty-six cases of primary malignant vascular tumors of the pleura were identified in the literature. In a total of 31 cases, 22 were from the West and 9 from Japan. Patients were 22 to 79 years old (average, 57), and the male/female ratio was 9:1. Prior chronic pyothorax was identified only in cases reported from Japan. History of exposure to radiation or asbestos was noted in a few Western cases. The most common presentation was pleural thickening and effusion. Almost all of the patients died of disease shortly after diagnosis. A spectrum of histology ranging from characteristic high-grade epithelioid to relatively low-grade EHE-like features was observed in our cases and can be found in previous reports. Most cases showed variable spotty cytokeratin immunoreactivity. Endothelial markers (factor 8, CD34, or CD31) were invariably positive. Pleural angiosarcomas are often epithelioid and can be easily mistaken for mesothelioma or carcinoma clinically and histologically. Awareness of this rare tumor should prompt the use of endothelial markers when faced with a questionable mesothelioma. When cytokeratin is negative, or focal with strong vimentin reactivity, a vascular tumor should be suspected and confirmed with vascular markers. Because of their invariably aggressive behavior, all epithelioid vascular tumors of the pleura should be considered highly malignant regardless of the presence of EHE-like histological features. PMID- 10665910 TI - HER-2/neu gene amplification by FISH predicts poor survival in Barrett's esophagus-associated adenocarcinoma. AB - The HER-2/neu oncogene is localized to chromosome 17q and shares significant homology with the epidermal growth factor receptor. HER-2/neu protein overexpression has been associated with poor prognosis in a variety of tumors, but its significance in Barrett's esophagus-associated adenocarcinoma (BEAd) is unknown. Therefore, the aim of this study was to evaluate the prevalence and prognostic value of HER-2/neu gene amplification by fluorescence in situ hybridization (FISH) in 63 cases of BEAd. Routinely processed tissue sections from resection specimens of 63 patients with BEAd (M/F ratio, 10:1; mean age, 63 years) were assayed for HER-2/neu gene amplification by FISH using the Ventana unique sequence probe (Ventana Medical Systems, Inc, Tuscon, AZ). FISH results were correlated with the pathological features of the tumors and with patient survival. Clinical follow-up data were available for 54 patients (mean follow-up, 31 months [range, 1 to 152 months]). The HER-2/ neu gene was amplified in 12 of 63 (19%) cases. The presence of HER-2/neu gene amplification showed a trend toward a correlation with depth of tumor invasion (P = .07), lymph node metastasis (P = .13), and pathological stage (P = .14), but did not correlate with any of the other pathological features, such as degree of differentiation or tumor size. On both univariate and multivariate analysis, HER-2/neu gene amplification was associated with shortened survival (P = .03). HER-2/neu oncogene amplification, as determined by FISH, correlates with shortened patient survival and independently predicts poor outcome in patients with BEAd. PMID- 10665911 TI - Orthotopic liver transplantation for familial amyloidotic polyneuropathy: a pathological study. AB - Familial amyloidotic polyneuropathy (FAP), a hereditary form of systemic amyloidosis with clinically significant neuropathy and cardiomyopathy, is caused by a genetic defect of the transthyretin gene, which is mostly synthesized in the liver. Orthotopic liver transplantation (OLT) is thought to eliminate the amyloidogenic protein and currently is the only definitive treatment for this disorder. The aim of this study was to define the distribution and extent of amyloid deposition in tissues from these patients and evaluate the suitability of the resected FAP livers for transplantation into non-FAP patients. Surgical specimens from 14 patients removed at the time of OLT and autopsy tissues from 3 of the 14 were examined histologically using hematoxylin and eosin and Congo red stained sections. The extent of amyloid deposits was evaluated, semiquantitatively graded from negative to marked, and correlated with clinical course and patient outcome. Amyloid deposits were consistently seen in hilar and vagus nerves. Liver lobular involvement was minimal in 1 and absent in the other 13 cases, with portal arterial amyloid deposits seen in 7 cases. At autopsy, extensive amyloid deposition in the heart was seen in all 3 cases with involvement of the conduction system. The extent of amyloid deposition at OLT did not correlate with the duration of symptoms before OLT or patient outcome after OLT. In conclusion, liver parenchymal involvement in FAP is minimal, and these explants are suitable for grafting in non-FAP patients. The recipients of such grafts must be carefully observed for the development of any amyloid-related disease, particularly cardiomyopathy. Of the tissues removed at OLT, the histopathologic confirmation of FAP is most consistently made by the examination of hilar and vagus nerves. PMID- 10665912 TI - Loss of p16/CDKN2A tumor suppressor protein in gastric adenocarcinoma is associated with Epstein-Barr virus and anatomic location in the body of the stomach. AB - Gastric adenocarcinomas (n = 125) were analyzed by immunohistochemistry for the presence of p16, the CDKN2A gene product. This protein was lost in 31 of 125 cases (25%), and loss was associated with location of the tumor in the body of the stomach (P = .001). Loss of p16 was also associated with the presence of Epstein-Barr virus (EBV) in tumor cells as determined by in situ hybridization (P = .022). This effect may relate to anatomic site, because EBV-associated tumors originate more frequently in the body of the stomach. When p16 status was evaluated for ethnic origin of the patient (non-Hispanic white, Hispanic, or black), a strong trend (P = .057) was found for African-American patients to have fewer p16-negative tumors than other patients. This also may relate to anatomic location, because fewer tumors from black patients arose in the body of the stomach (P = .022). No significant associations were detected between p16 status and histological subtype (intestinal v diffuse), the presence of microsatellite instability, grade or stage of the tumor, or age, gender, or survival of the patient. In conclusion, p16 loss is quite common in gastric adenocarcinoma, and such loss is more common in EBV-infected tumors arising in the body of the stomach. PMID- 10665913 TI - Morphometric analysis of the "mucocellular layer" overlying colorectal cancer and normal mucosa: relevance to exfoliation and stool screening. AB - Characterization of shed cell elements entrapped within the colorectal surface mucus would be valuable to the study of exfoliation and candidate stool screening markers. Yet, surprisingly little is known about the cellular composition of this "mucocellular layer" (MCL). Our aim was to describe and compare the histomorphometry of the MCL that overlies colorectal cancer (CRC) and normal mucosa. From tissue archives, 20 resected CRC specimens yielding perpendicular cuts of both tumor surface and adjacent normal mucosa were consecutively selected. MCL thickness and cell number were determined in triplicate using an ocular micrometer. Cellular elements within the MCL were characterized on paraffin sections by immunohistochemistry. Mean cell density was much greater in the MCL over CRC (2,639 +/- 2,178 per mm2) than over normal mucosa (184 +/- 395 per mm2), p < .001. Robust-appearing colonocytes and inflammatory cells predominated in the hypercellular MCL of CRC; the former retained expression of tumor-associated antigens. In contrast, the sparsely scattered cells within the normal MCL were typically apoptotic and of indeterminate lineage. Based on direct observations from this first descriptive study of the colorectal MCL, luminal shedding appears to be much greater from CRC than from normal mucosa. PMID- 10665914 TI - Differential expression of thyroid transcription factor 1 in small cell lung carcinoma and Merkel cell tumor. AB - The distinction between metastatic small cell lung carcinoma (SCLC) and Merkel cell tumor is difficult by routine histology, prompting the search for specific markers that could separate these neoplasms. Thyroid transcription factor 1 (TFF 1) is a homeodomain containing transcription factor expressed in the normal airway epithelium. The expression of TTF-1 has also been shown in adenocarcinomas and small cell carcinomas of the lung. However, the utility of TTF-1 to differentiate between SCLC and Merkel cell tumor has not yet been investigated. In this study, paraffin sections of 36 SCLCs and 21 Merkel cell tumors were analyzed for the presence of immunoreactive TTF-1 and cytokeratin 20 (CK20), a marker previously demonstrated in Merkel cell tumors. Monoclonal TTF-1 and CK20 antibodies were used with a biotin-streptavidin detection system. Immunostaining for TTF-1 was observed in 97% of SCLCs and in no Merkel cell tumors. Immunoreactivity for CK20 was demonstrated in 76% of Merkel cell tumors and 3% of SCLCs. These data indicate that TTF-1 is a sensitive (97%) and specific (100%) marker for SCLCs and can be used to differentiate SCLCs from Merkel cell tumors. PMID- 10665915 TI - Solitary fibrous tumor of the lower urogenital tract: a report of five cases involving the seminal vesicles, urinary bladder, and prostate. AB - Solitary fibrous tumor (SFT) is not as site-restricted as once believed. Initially described as a tumor of the pleura, SFT is now recognized at various extrathoracic sites. We report 5 cases of extrapleural SFT involving the male lower urogenital tract. The tumors involved the seminal vesicles (2 cases), urinary bladder (2 cases), and the prostate (1 case). The patients with bladder tumors were asymptomatic. The patients with seminal vesicle involvement presented with hematuria or groin pain. The patient with prostate involvement presented with urinary retention. The prostate tumor was large, and it could not be completely excised because of its extensive spread beyond the prostate into the pelvis. The other 4 tumors were completely excised, and none has recurred during limited follow-up. By histological criteria, 4 of the tumors were benign, and 1 was malignant. Even though the classic histological features of SFT were well developed in each case, all 5 tumors were initially misdiagnosed, including 3 benign tumors that were misclassified as sarcomas. These 5 cases confirm the male lower genitourinary tract as yet another site of origin for SFTs, challenge the notion that extrapleural SFTs invariably are benign, and draw attention to the problem of recognizing SFTs when they arise in unexpected sites. PMID- 10665916 TI - Impact of human immunodeficiency virus infection on the histological features of chronic hepatitis C: a case-control study. The MULTIVIRC group. AB - Hepatitis C virus (HCV) is frequently encountered in human immunodeficiency virus (HIV)-infected patients because of common routes of transmission. Previous studies suggested that HIV infection impaired the natural course of chronic hepatitis C, with a more rapid progression to cirrhosis. However, these studies did not assess the HIV infection impact on chronic hepatitis C by taking into account the risk factors for liver fibrosis progression: alcohol, sex, age at the contamination, and duration of HCV infection. We studied liver biopsy specimens of 2 groups of 58 patients that were infected by both HCV and HIV or by HCV alone. The 2 groups were matched according those risk factors, and liver biopsy responses were evaluated with the METAVIR items. The METAVIR activity was higher in HIV-positive than HIV-negative patients. Cirrhosis was more frequent: (1) in HIV-positive patients with CD4 < or = 200 cells/microL (45%) than in HIV-negative patients (10%) (P = .003), (2) in HIV-positive patients with CD4 < or = 200 cells/microL (45%) than in HIV-positive patients with CD4 > 200 cells/microL (17%) (P = .04). These differences, which were linked to HIV status, might be related to the enhanced HCV replication during HIV infection or other immune mechanisms that need further studies. PMID- 10665917 TI - The histological spectrum of visceral leishmaniasis caused by Leishmania infantum MON-1 in acquired immune deficiency syndrome. AB - Visceral leishmaniasis (VL) due to Leishmania infantum is endemic in Southern France and can be considered as an opportunistic infection in patients with acquired immunodeficiency syndrome (AIDS). Co-infection with Leishmania sp. and human immunodeficiency virus (HIV) is emerging, but pathological findings of leishmaniasis in AIDS have been poorly documented, and scattered case reports have include morphological descriptions. The clinicopathologic analysis of 16 patients with HIV and VL were evaluated. The clinical presentation was characteristic of VL, with fever, hepatosplenomegaly, and pancytopenia in 6 patients, and the diagnosis was confirmed by finding amastigotes of Leishmania sp. in bone marrow smears and biopsy specimens. In 4 patients, the initial diagnosis of VL was made fortuitously in gastrointestinal biopsies performed systematically (3 patients) or in case of diarrhea (1 patient). In one duodenal biopsy, Leishmania sp. and Mycobacteria sp. were associated. Liver biopsy allowed the diagnosis of VL in 3 cases. Autopsy was performed in 9 patients, showing a disseminated leishmaniasis with very unusual localizations (adrenal and heart) in 2 cases. Cutaneous leishmaniasis involvement was noted before (4 patients), at the same time (2 patient), or after (1 patient) the diagnosis of VL. Inflammatory infiltrates noted with Leishmania sp. infection were made by CD68 macrophages with (8 patients) or without (8 patients) associated CD8 positive lymphocytes. Immunoperoxidase study using polyclonal anti-Leishmania sp. antibodies contributed to the diagnosis in all cases. Electron microscopy of 2 digestive biopsy specimens showed the ultrastructural characteristics of Leishmania sp. amastigotes. The zymodeme MON-1 of L infantum was identified by isoenzyme electrophoresis in all patients. The mean of CD4 counts was 37/mm3 at the time of diagnosis, and the mean duration before the death was 8 months. As shown in this study, VL in AIDS can be diagnosed in gastrointestinal or liver biopsies. Diagnosis of VL was made when the CD4 count was very low and was correlated with a poor prognosis. PMID- 10665918 TI - Placental pathology in malaria: a histological, immunohistochemical, and quantitative study. AB - To characterize the histological changes in malarial placentas and their relationship with parity and maternal and cord parasitemias, we conducted a histological study on 1,179 placentas from Ifakara, Tanzania, an area with intense and perennial malaria transmission. Immunohistochemical and quantitative studies for CD45, fibrin, and villous area were performed in 60 cases. Four hundred fifteen placentas (35.2%) showed parasites (active infections); in 303 of them, parasites co-existed with pigment covered by fibrin (chronic infections), and in 112 only parasites were detected (acute infections). Four hundred seventy five cases (40.3%) showed hemozoin deposition without parasites (past infections). Of women with parasitized placentas, 46.3% did not show parasites in the peripheral blood. Basal membrane thickening (P = .002), fibrinoid necrosis (P = .004), and prominence of syncytial knots (P = .031) were associated with active malarial infection. No quantitative differences for perivillous fibrin deposition or villous area were found. The most significant association with active malarial infection was intervillous infiltration by mononuclear inflammatory cells (P < .001). Chronic infections were associated with the most severe changes, particularly intervillous mononuclear inflammation (OR, 28.7; 95% CI = 16.0 to 51.5, P< .001). Past infections showed only minimal differences with noninfected placentas. Primiparas showed chronic infections more frequently than multiparas (52% v 15%, P < .001). They also showed significantly higher placental parasitemias and intervillous inflammatory infiltrate. In conclusion, placental histology is more sensitive than peripheral blood examination in detecting malarial infection during pregnancy. Most malarial infections recover during pregnancy, leaving few residual changes in the placenta. Intervillous inflammation is the most frequent finding associated with malaria and is especially severe in primiparas, suggesting that mechanisms other than immunosuppression are responsible for the high susceptibility in this group. PMID- 10665919 TI - Luminal contents of benign and malignant prostatic glands: correspondence to altered secretory mechanisms. AB - Recent changes in tissue fixation strategy, using glutaraldehyde, have clarified the secretory mechanisms of the normal prostate identifying cytoplasmic prostatic secretory granules, structures not preserved by formalin fixation. This normal secretory mechanism was absent in most adenocarcinomas, depicting an important metabolic change in transformed prostate cells. The current study further investigates differences between benign and malignant prostate secretion and relates them to the production of corpora amylacea by benign glands and crystalloids or mucin by cancer. In all normal prostate cells examined (6 cases), prostate secretory granules (PSG) were approximately 1-microm, brightly eosinophilic granules filling the cytoplasm of secretory cells and released in packets by a specialized apocrine cell structure. After apocrine decapitation and luminal dispersal, some of the cytoplasmic and PSG remnants condensed to form eosinophilic bodies (EB) with a glycoprotein rim and central protein core. EB were observed adsorbing and layering onto the surface of prostatic corpora amylacea representing their chief mode of enlargement. Biochemical analysis and x ray diffraction studies confirmed sulfated glycosaminoglycans of similar structure as the main constituent of both PSG and corpora amylacea. Peripheral zone amphiphilic "dark cell" carcinoma (9 cases) contained almost no PSG, and showed neither apical decapitation nor EB formation, but mucin secretion was frequently detected. Crystalloids that share the same staining characteristics and sulfur content as PSG and corpora amylacea were identified in 3 selected "clear cell" carcinomas, all of which showed at least focal PSG secretion. The recognition of these differing secretory mechanisms and their deviation from normal further defines the histological criteria and spectrum of prostate malignancy. PMID- 10665920 TI - Recurrent hepatitis B, hepatitis C, and combined hepatitis B and C in liver allografts: a comparative pathological study. AB - Although recurrence of viral hepatitis in liver transplants is common, data comparing recurrent hepatitis B (HB), hepatitis C (HC), and co-existing dual hepatitis B and C (HB&C) are sparse. Posttransplantation liver biopsies, along with molecular, serological, immunohistochemical, and clinical data from 27 patients with pretransplantation diagnosis of chronic viral hepatitis, were reviewed. The patients were placed into 4 groups: Group I, with pretransplantation HB (n = 8); group II, with pretransplantation HC (n = 10); group III with pretransplantation HC and anti-HB surface or core antibody (n = 4); and group IV, with pretransplantation HB&C (n = 5). The histopathologic findings and patient outcome were compared in the 4 groups. A high rate of recurrence of viral hepatitis was seen for all 4 groups: Group I = 100%, group II = 90%, Group III = 100%, and group IV = 80%, with the mean (median) recurrence time of 308 (224), 82 (52), 61 (64), and 125 (70) days, respectively. The number of deaths (their median survival times) were: group I = 4 (374 days), group II = 4 (794 days), group III = 1 (1,143 days), and group IV = 5 (448 days). The earliest histological findings of lobular injury was the presence of acidophil bodies and Kupffer cell hyperplasia, the latter being more prominent in recurrent HC cases. Recurrent HB presented in 2 forms: early (before 150 days) with poor survival and with either severe necroinflammatory histology or with features of fibrosing cholestatic hepatitis, and delayed (after 150 days), with mild necro inflammatory activity and prolonged survival. HC with or without anti-HB antibodies had early recurrence, but the course was slowly progressive. Patients with HB&C had recurrence of both viruses; however, the course was dictated by HB virus. PMID- 10665921 TI - Chromosomal alterations in ulcerative colitis-related and sporadic colorectal cancers by comparative genomic hybridization. AB - Both ulcerative colitis (UC)-related and sporadic colorectal cancers are thought to evolve through a multistep process of genomic instability, accumulation of genomic alterations, and clonal expansion. This process may involve different genomic changes in UC-related cancers than in sporadic cancers because of the origin of UC-related cancers in an inflammatory field. This study was designed to define the specific genomic events occurring in UC-related cancers. Comparative genomic hybridization (CGH) was performed on 32 UC-related and 42 stage-matched sporadic colorectal cancers. The mean number of chromosomal alterations per case was similar in the UC-related and sporadic tumor groups (8.6 in UC, 8.1 in sporadic). The 2 tumor groups shared many chromosomal alterations: losses on 18q (78% UC v69% sporadic), 8p (53% v50%), 17p (44% v57%), and gains on 8q (63% v45%), 20q (44% UC v67%), and 13q (44% UC v38%). However, differences in the frequency and timing of specific alterations were observed. Chromosome 5q was lost in 56% of UC-related but in only 26% of sporadic cancers. Alterations of chromosome 8 were associated with stage progression in UC-related, but not in sporadic cancers. In contrast, 18q loss was associated with stage progression in sporadic cancers only. Thus, differences in the frequency and timing of individual chromosomal alterations suggest that genetic progression in these 2 tumor groups may follow multiple pathways. PMID- 10665922 TI - Incidence of p14ARF gene deletion in high-grade adult and pediatric astrocytomas. AB - The INK4a-ARF locus encodes 2 separate proteins through differential splicing of alternative first exons to produce p16INK4a (exon 1alpha) and p14ARF (exon 1beta) products in human cells. The p16INK4a protein inhibits the cyclin D-dependent kinases (CDK) that control the phosphorylation of the Rb protein and cell proliferation. The p14ARF gene product can complex with and sequester the MDM2 protein within the nucleus, thus modulating the activity of the p53 protein. Loss of p16INK4a expression would disrupt the retinoblastoma (Rb)/p16INK4a/cyclin D dependent kinase (CDK4) pathway, whereas loss of p14ARF expression would inactivate both the Rb and p53/ MDM2/p14ARF pathways through MDM2, which can complex with either Rb or p53. Loss of the p16INK4a gene on 9p21 has been documented in a wide range of human tumors, including one third of glioblastomas. However, in tumors showing homozygous loss of exon 2 of the p16INK4a gene, loss of exon 1beta of the p14ARF gene has not been established. In this study, we have assessed deletion of the p14ARF gene in 29 pediatric and 107 adult high-grade astrocytomas and 9 glioma cell lines, using multiplex PCR analysis for exon 1beta. We found homozygous deletions for exon 1alpha and exon 1beta in 3 of 29 (10%) of the pediatric cases (2 grade III, 1 grade IV), 25 of 107 (23%) of the adult cases (6 grade III and 19 grade IV), and 8 of 9 (89%) of the glioma cell lines. Therefore, loss of the INK4a-ARF locus in high-grade astrocytomas may contribute to the highly malignant behavior and treatment resistance of these tumors through elimination of multiple checkpoint cell cycle control proteins. PMID- 10665923 TI - Chagas' disease of the cervix uteri in a patient with acquired immunodeficiency syndrome. AB - A 27-year-old woman with the acquired immunodeficiency syndrome and endocervical Chagas' disease is reported. The patient was a cocaine addict, and her sexual partner was also human immunodeficiency virus (HIV)-positive. Her past medical history included a son who died 3 days after birth due to congenital Chagas' disease. Seven years later, through a cervical biopsy, a reactivation of Chagas' disease was diagnosed. Giant cells with typical amastigotes were seen; they strongly stained with antibodies against Trypanosoma cruzi. The patient died 5 months later of an acute chagasic myocardiopathy. To our knowledge, this is the first report of this parasitosis in the cervix uteri. PMID- 10665924 TI - Nodular glomerulosclerosis secondary to mu heavy chain deposits. AB - mu heavy chain deposition disease is very rare. We report the first case of glomerulonephritis in a woman without evidence of hematopoietic malignancy. Nodular glomerulosclerosis and monotypic mu heavy chain mesangial deposits were identified by immunofluorescence without kappa or lambda deposits. Electron microscopy showed fibrillar mesangial deposits of 16-18 nm in diameter. Serum immunoglobulins, cryoglobulins, serum immunoelectrophoresis, and immunofixation, bone marrow biopsy, and Bence Jones proteins in urine were negative. The patient has stable renal disease and is free of malignancy 6 years after the initial occurrence of proteinuria. PMID- 10665925 TI - Benign metastasizing leiomyoma: a cytogenetically balanced but clonal disease. AB - Benign metastasizing leiomyoma (BML) is a rare condition, characterized by the occurrence of multiple smooth-muscle nodules, most often located in the lung after previous hysterectomy because of histologically benign appearing leiomyoma. Although the condition resembles a metastatic process, case studies provided evidence that it may be the result of an intravenous leiomyomatosis or an independent and multifocal smooth-muscle proliferation. Comparative genomic hybridization and X-chromosome inactivation analysis were used in a case of BML to determine whether pulmonary and uterine tumors are related one to another. A balanced karyotype, previously reported in leiomyomas and an identical X chromosome inactivation pattern found in all tumorlets, is most consistent with a monoclonal origin of both uterine and pulmonary tumors and the interpretation that pulmonary lesions are metastatic. PMID- 10665926 TI - Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules. AB - The receptor-associated protein (RAP) has been identified as a chaperone regulating the expression and processing of the LDL receptor-related protein. RAP also binds to the related 600-kD multiligand endocytic receptor megalin expressed in many absorptive epithelia including renal proximal tubule. The present study examines the effect of RAP gene disruption on megalin expression and subcellular distribution in the proximal tubule as well as the effect on tubular protein reabsorption. It is shown that RAP is important for the normal expression and function of megalin. Megalin expression was reduced to approximately 23% estimated by immunoblotting and supported by immunocytochemistry and by the amount of megalin recovered by RAP affinity chromatography. Light- and electron microscope immunocytochemistry as well as analyses on separated membrane fractions showed significant changes in the subcellular distribution of megalin. A significant reduction in the normal brush border labeling was observed in association with increased labeling of rough endoplasmic reticulum and the smooth paramembranous endoplasmic reticulum along the basolateral membranes. RAP deficiency was associated with changes in urinary protein composition, enabling the identification of alpha-amylase as a new ligand for megalin. In addition, an increased excretion of vitamin D-binding protein, a recently identified ligand to megalin, was observed supporting changes in tubular protein reabsorption. The present data show that RAP is of crucial importance for normal processing and function of megalin, suggesting a chaperone-like function of this protein in the kidney proximal tubule. PMID- 10665927 TI - Effect of chronic metabolic acidosis on calbindin expression along the rat distal tubule. AB - Calbindin D28k has been reported to be involved in the transcellular calcium transport along the rat distal tubule. It has also been shown that chronic metabolic acidosis (CMA) induces significant hypercalciuria. The present study investigated whether CMA affects the mRNA and the protein expression of calbindin D28k along isolated distal tubule (DT) of rats. The animals were made acidotic by adding 0.28 mol/L NH4Cl to the drinking water for 7 d. This maneuver was associated with an increase in plasma ionized calcium. Inulin clearance experiments demonstrated that metabolic acidosis did not affect GFR, but it significantly increased both total and fractional urinary calcium excretion. To define the role of calbindin D28k, total RNA was extracted from DT, identified, and microdissected from collagenase-treated kidneys. cDNA was synthesized from RNA using reverse transcriptase and oligo(dT)(12-18) primers. Calbindin D28k mRNA abundance was semiquantified by a competitive reverse transcription-PCR, using an internal standard of cDNA that differed from the wild-type calbindin D28k by a deletion of 86 bp. The reverse transcription-PCR was performed starting from the same amount of total RNA. For each set of experiments, control and acidotic rats were studied in parallel. The identity of the DT was further verified by the presence of the thiazide-sensitive NaCl cotransporter (rTSC1) mRNA. Calbindin D28k mRNA abundance was 0.89 +/- 0.21 amol/ng total RNA in DT of CMA rats (n = 5) compared with 0.30 +/- 0.12 amol/ng total RNA of control rats (n = 5) (P < 0.05). Using specific rabbit polyclonal anti-calbindin D28k antibody, Western blotting was performed starting from thin slices of outer cortex. Densitometric analysis revealed that in acidotic rats (n = 7) there was a 17 +/- 5% (P < 0.05) increase in calbindin D28k protein abundance compared with controls (n = 7). These results indicate that in the rat, ammonium chloride loading induces an increase in filtered ionized calcium load that is associated with a significant upregulation of calbindin D28k both at the mRNA and protein level. These last effects will help to reduce the concomitant hypercalciuria, thus mitigating the consequence of CMA on calcium metabolism. PMID- 10665928 TI - Ischemic acute renal failure induces differential expression of small heat shock proteins. AB - AlphaB-crystallin and heat shock protein (hsp) 25 are structurally and functionally related small stress proteins induced by a variety of insults, including heat and ischemia. Cytoprotection by these two hsp is thought to result from molecular chaperoning and/or cytoskeletal stabilization. Because renal ischemia is characterized by disruption of the renal tubular cell actin cytoskeleton, this study was conducted to determine the localization and quantify the expression and phosphorylation of both hsp in renal cortex, isolated glomeruli, outer medulla, and inner medulla of rats after bilateral renal ischemia. Sham-operated kidneys had similarly small amounts of hsp25 and alphaB crystallin in cortex and glomeruli, with substantially greater amounts of alphaB crystallin versus hsp25 in outer and inner medulla. Ischemia resulted in significantly increased hsp25 (and hsp70i) but variable alphaB-crystallin levels in cortex and outer medulla, and progressively decreased glomerular hsp25 phosphorylation. In sham-operated kidneys, hsp25 localized to glomeruli, vessels, and collecting ducts, with alphaB-crystallin primarily in medullary thin limbs and collecting ducts. After ischemia, hsp25 accumulated in proximal tubules in cortex and outer medulla, while alphaB-crystallin labeling became nonhomogeneous in outer medulla, and increased in Bowman's capsule. It is concluded that: (1) There is striking differential expression of hsp25 and alphaB-crystallin in various renal compartments; and (2) Renal ischemia results in differential accumulation of hsp25 and alphaB-crystallin, with hsp25 part of a generalized stress response in renal proximal tubular cells, which may play a role in recovery from ischemia-induced actin filament disruption. PMID- 10665929 TI - Parathyroid hormone stimulates extracellular signal-regulated kinase (ERK) activity through two independent signal transduction pathways: role of ERK in sodium-phosphate cotransport. AB - Parathyroid hormone (PTH), a major physiologic regulator of proximal renal tubule cell sodium-phosphate cotransport, stimulates several signal transduction pathways including extracellular signal-regulated kinases (ERK). The physiologic role of PTH-stimulated ERK is unknown. The purpose of the present study was to identify signaling components involved in PTH-stimulated ERK activity and to determine the role of PTH-stimulated ERK activity in regulation of phosphate transport. PTH-stimulated ERK activity was measured in opossum kidney (OK) cell lysates as phosphorylation of myelin basic protein by an in vitro kinase assay. PTH stimulated a dose-dependent increase in ERK activity with a peak at 10(-7) M. The time course was biphasic with an early peak at 10 min and a later peak at 20 min. Pretreatment of OK cells with the nonreceptor tyrosine kinase inhibitors genistein and herbimycin A or with the phosphatidylinositol 3-kinase (PI-3K) inhibitors wortmannin and LY294002 blocked the early and late peaks of PTH stimulated ERK activity. Pretreatment with the protein kinase C inhibitor calphostin C blocked only the later phase of PTH-stimulated ERK. To determine the role of ERK in regulation of phosphate transport, PTH inhibition of phosphate uptake and PTH regulation of sodium-phosphate cotransporter (NaPi-4) expression were measured in OK cells pretreated with the MEK inhibitor PD098059. PD098059 significantly attenuated PTH inhibition of phosphate uptake but did not prevent PTH downregulation of NaPi-4. It is concluded that PTH stimulates ERK through two signal transduction pathways: an early pathway dependent on tyrosine kinase and PI-3K and a late pathway dependent on protein kinase C. PTH-stimulated ERK regulates phosphate transport by a mechanism other than downregulation of NaPi-4 expression. PMID- 10665930 TI - Differential activation of mitogen-activated protein kinases in experimental mesangioproliferative glomerulonephritis. AB - Multiple extracellular mitogens are involved in the pathogenesis of proliferative forms of glomerulonephritis (GN). In vitro studies demonstrate the pivotal role of mitogen-activated protein (MAP) kinases in the regulation of cellular proliferation. This study was conducted to examine whether these kinases, as a convergence point of mitogenic stimuli, are activated in mesangioproliferative GN in vivo. Therefore, anti-Thy1 GN was induced in rats using a monoclonal anti Thy1.1 antibody (OX-7). Whole cortical tissue as well as isolated glomeruli were examined at different time points using kinase activity assays and Western blot analysis. A maximal increase in the number of glomerular mitotic figures (9.7 fold) was demonstrated 6 d after injection of the anti-Thy1.1 antibody. In parallel with this finding, a significant increase in cortical, and more dramatically glomerular, activity of extracellular signal-regulated kinase (ERK) was detected. Maximal activation of ERK was detectable on day 6. This activation of ERK was accompanied by an increase in the expression of MEK (MAP kinase/ERK kinase), the ERK-activating kinase. A marked induction of glomerular apoptosis at 2 h after injection of the anti-Thy1.1 antibody, which subsided subsequently, was demonstrated using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay as well as staining for single-stranded DNA. However, no significant activation of stress-activated protein kinase or p38 MAP kinase, both MAP kinases that are suggested to induce apoptosis and to inhibit cellular growth, was detectable at this early time point. Rather, on day 6 a dramatic decrease in the activity of p38 MAP kinase, which might have contributed to the overshooting glomerular cellular proliferation, was observed. Treatment of rats with heparin blunted glomerular proliferation as well as ERK activation and restored p38 MAP kinase activity. These observations point to ERK and p38 MAP kinase as putative mediators of the proliferative response in mesangioproliferative GN and suggest that upregulation of MEK is involved in the long-term regulation of ERK in vivo. PMID- 10665931 TI - Absence of CD89, polymeric immunoglobulin receptor, and asialoglycoprotein receptor on human mesangial cells. AB - IgA nephropathy (IgAN) is characterized by raised serum IgA and predominant mesangial IgA deposits of polymeric nature. The expression of IgA receptor molecules in white blood cells and glomerular mesangial cells has recently attracted much attention in relation to the uptake of IgA by these cells. This study investigates the expression of IgA Fc receptor (Fc alphaR1 or CD89), asialoglycoprotein receptor (ASGPR), and polymeric Ig receptor (pIgR) in cultured glomerular mesangial cells. Using a sensitive nested reverse transcription-PCR, mRNA encoding for Fc alphaR1, pIgR, or the H2 chain of ASGPR was not demonstrated on human mesangial cells. U937, HepG2, and HT29 cell lines, used as positive controls, strongly expressed the Fc alphaR1, ASGPR, and pIgR mRNA, respectively, under similar experimental conditions. Flow cytometry also demonstrated the presence of surface proteins for Fc alphaR1, ASGPR, and pIgR on the respective control cell lines but not on human mesangial cells. Expression of Fc alphaR1 mRNA on cultured U937 cells was upregulated by tumor necrosis factor-alpha. However, tumor necrosis factor-alpha, interleukin-1beta, or transforming growth factor-beta failed to induce the expression of Fc alphaR1 on human mesangial cells. Human serum IgA or secretory IgA bound to human mesangial cells, HepG2, or the U937 cell line in a dose-dependent manner. The binding of purified IgA to human mesangial cells was not blocked by preincubation with human IgG, IgM, orosomucoid, asialo-orosomucoid, anti-CD89 antibody (My43), or anti-secretory component antibody. The present study concluded that there was an absence of Fc alphaR1, ASGPR, or pIgR on human mesangial cells. These findings suggest that the predominant binding of human IgA to human mesangial cells is mediated by other mechanisms. PMID- 10665932 TI - Central role for interferon-gamma receptor in the regulation of renal MHC expression. AB - The role of the interferon-gamma (IFN-gamma) receptor 1 (IFN-gammaR1) was investigated in the regulation of MHC expression in kidney in the basal state, in response to potent inflammatory stimuli, and after renal injury. In this study, MHC regulation in mice lacking IFN-gammaR due to targeted disruption of the IFN gammaR1 gene (GRKO mice) was compared with regulation in 129Sv/J mice with wild type IFN-gammaR1 genes. Basal class I expression was reduced by approximately 45% in kidneys of GRKO mice, while basal class II expression was confined to interstitial cells and was not reduced in GRKO kidneys. Recombinant IFN-gamma administration induced widespread expression of class I and II in renal tubules, arterial endothelium, and glomeruli of 129Sv/J mice, but produced no change in kidneys of GRKO mice. Potent systemic inflammatory stimuli (injections of allogeneic cells, skin sensitization with oxazolone, and injection of bacterial lipopolysaccharide) significantly induced both class I and class II expression in 129Sv/J mice, but not in GRKO mice. Acute renal injury increased local expression of class I and II in both 129Sv/J and GRKO mice, but the induction in GRKO mice was reduced compared with 129Sv/J mice. Thus, the IFN-gamma receptor plays a unique and nonredundant role in the regulation of renal MHC in the response to inflammation, in the response to renal injury, and in the basal state. PMID- 10665933 TI - Interleukin-10 inhibits macrophage-induced glomerular injury. AB - The ability of interleukin-10 (IL-10) to inhibit macrophage recruitment, activation, and proliferation in vivo was studied in a macrophage-mediated, but T cell-independent, passive anti-glomerular basement membrane antibody-induced model of glomerulonephritis (GN) in rats. Treatment with recombinant murine IL-10 resulted in dose-dependent reductions in proteinuria (high dose: 16 +/- 1 mg/24 h; low dose: 30 +/- 2 mg/24 h; control treatment: 69 +/- 6 mg/24 h; normal: 7 +/- 1 mg/24 h) and glomerular macrophage recruitment (high dose: 1.8 +/- 0.1 macrophages per glomerular cross section [c/gcs]; low dose: 5.5 +/- 0.2 c/gcs; control treatment: 12.1 +/- 0.6 c/gcs). Macrophage and intrinsic glomerular cell proliferation were reduced at both doses of IL-10, as was glomerular expression of P-selectin and monocyte chemoattractant protein-1. IL-10 treatment also resulted in a dose-dependent reduction of macrophage activation as indicated by MHC class II and IL-1beta expression. Glomerular nitrite production by isolated cultured glomeruli was reduced after IL-10 treatment in vivo (high dose: 2.3 +/- 2.3 nmol/10(4) glomeruli per 72 h; low dose: 28 +/- 5 nmol/10(4) glomeruli per 72 h; control treatment: 82 +/- 11 nmol/10(4) glomeruli per 72 h). Tumor necrosis factor-alpha production was abolished by high-dose treatment and reduced by the lower dose (3.8 +/- 3.8 pg/10(4) glomeruli per 72 h; control treatment: 249 +/- 23 pg/10(4) glomeruli per 72 h). These studies demonstrate that IL-10 directly attenuates glomerular macrophage recruitment, activation, and proliferation in vivo and can significantly attenuate macrophage-mediated GN independent of any effects on T cells. PMID- 10665934 TI - Nephrocystin: gene expression and sequence conservation between human, mouse, and Caenorhabditis elegans. AB - Juvenile nephronophthisis, an autosomal recessive cystic kidney disease, is the primary genetic cause for chronic renal failure in children. The gene (NPHP1) for nephronophthisis type 1 has recently been identified. Its gene product, nephrocystin, is a novel protein of unknown function, which contains a src homology 3 domain. To study tissue expression and analyze amino acid sequence conservation of nephrocystin, the full-length murine Nphp1 cDNA sequence was obtained and Northern and in situ hybridization analyses were performed for extensive expression studies. The results demonstrate widespread but relatively weak NPHP1 expression in the human adult. In the adult mouse there is strong expression in testis. This expression occurs specifically in cell stages of the first meiotic division and thereafter. In situ hybridization to whole mouse embryos demonstrated widespread and uniform expression at all developmental stages. Amino acid sequence conservation studies in human, mouse, and Caenorhabditis elegans show that in nephrocystin the src-homology 3 domain is embedded in a novel context of other putative domains of protein-protein interaction, such as coiled-coil and E-rich domains. It is concluded that for multiple putative protein-protein interaction domains of nephrocystin, sequence conservation dates back at least to Caenorhabditis elegans. The previously described discrepancy between widespread tissue expression and the restriction of symptoms to the kidney has now been confirmed by an in-depth expression study. PMID- 10665935 TI - Combined mycophenolate mofetil and losartan therapy arrests established injury in the remnant kidney. AB - Previously it was shown that early treatment with mycophenolate mofetil (MMF) attenuated renal inflammation, glomerulosclerosis (GS), and interstitial expansion in the 5/6 ablation (NX) model. Angiotensin II antagonists also mitigate renal injury in NX, presumably by lowering glomerular pressure (P(GC)). This study investigated: (1) whether combined MMF/angiotensin II antagonists treatment affords superior protection compared with the respective monotherapies; and (2) whether this association is effective even when instituted late in the course of the disease. Adult male Munich-Wistar rats underwent NX, remaining untreated for 30 d. BP, albuminuria, and the extent of GS, interstitial expansion, and macrophage infiltration were then determined in 17 rats. The remaining 118 rats received either inert vehicle or one of the following: MMF, 10 mg/kg by gavage once daily; losartan potassium (L), 20 mg/dl in drinking water; or combined MMF/L treatment. Sixty days after ablation, untreated NX rats exhibited marked glomerular hypertension, which was attenuated by MMF and, more effectively, by either L or combined MMF/L treatment. At 120 d, hypertension and albuminuria were worsened in untreated NX rats, which exhibited intense macrophage infiltration and severe glomerular and interstitial disease. L and, to a lesser extent, MMF monotherapies attenuated these abnormalities, without preventing their progression. In rats given combined MMF/L therapy, macrophage infiltration, GS, and interstitial expansion remained at pretreatment levels. By acting on two distinct pathogenic mechanisms, combined MMF/L treatment arrested established renal injury in the NX model. Further investigation is needed to determine whether this association can prevent renal scarring in other models and in human disease. PMID- 10665936 TI - A mouse model of renal tubular injury of tyrosinemia type 1: development of de Toni Fanconi syndrome and apoptosis of renal tubular cells in Fah/Hpd double mutant mice. AB - Hereditary tyrosinemia type 1 (HT1) (McKusick 276700), a severe autosomal recessive disorder of tyrosine metabolism, is caused by mutations in the fumarylacetoacetate hydrolase gene Fah (EC 3.7.1.2), which encodes the last enzyme in the tyrosine catabolic pathway. HT1 is characterized by severe progressive liver disease and renal tubular dysfunction. Homozygous disruption of the gene encoding Fah in mice causes neonatal lethality (e.g., lethal Albino deletion c14CoS mice), an event that limits use of this animal as a model for HT1. A new mouse model was developed with two genetic defects, Fah and 4 hydroxyphenylpyruvate dioxygenase (Hpd). The Fah-/- Hpd-/- mice grew normally without evidence of liver and renal disease, and the phenotype is similar to that in Fah+/+ Hpd-/- mice. The renal tubular cells of Fah-/- Hpd-/- mice, particularly proximal tubular cells, underwent rapid apoptosis when homogentisate, the intermediate metabolite between HPD and FAH, was administered to the Fah-/- Hpd-/- mice. Simultaneously, renal tubular function was impaired and Fanconi syndrome occurred. Apoptotic death of renal tubular cells, but not renal dysfunction, was prevented by pretreatment of the animals with YVAD, a specific inhibitor of caspases. In the homogentisate-treated Fah-/- Hpd-/- mice, massive amounts of succinylacetone were excreted into the urine, regardless of treatment with inhibitors. It is suggested that apoptotic death of renal tubular cells, as induced by administration of homogentisate to Fah-/- Hpd-/- mice, was caused by an intrinsic process, and that renal apoptosis and tubular dysfunctions in tubular cells occurred through different pathways. These observations shed light on the pathogenesis of renal tubular injury in subjects with FAH deficiency. These Fah-/- Hpd-/- mice can serve as a model in experiments related to renal tubular damage. PMID- 10665937 TI - Effects of tetrahydrobiopterin on endothelial dysfunction in rats with ischemic acute renal failure. AB - The role of nitric oxide (NO) in ischemic renal injury is still controversial. NO release was measured in rat kidneys subjected to ischemia and reperfusion to determine whether (6R)-5,6,7,8-tetrahydro-L-biopterin (BH4), a cofactor of NO synthase (NOS), reduces ischemic injury. Twenty-four hours after bilateral renal arterial clamp for 45 min, acetylcholine-induced vasorelaxation and NO release were reduced and renal excretory function was impaired in Wistar rats. Administration of BH4 (20 mg/kg, by mouth) before clamping resulted in a marked improvement of those parameters (10(-8) M acetylcholine, delta renal perfusion pressure: sham-operated control -45 +/- 5, ischemia -30 +/- 2, ischemia + BH4 -43 +/- 4%; delta NO: control +30 +/- 6, ischemia + 10 +/- 2, ischemia + BH4 +23 +/- 4 fmol/min per g kidney; serum creatinine: control 23 +/- 2, ischemia 150 +/- 27, ischemia + BH4 48 +/- 6 microM; mean +/- SEM). Most of renal NOS activity was calcium-dependent, and its activity decreased in the ischemic kidney. However, it was restored by BH4 (control 5.0 +/- 0.9, ischemia 2.2 +/- 0.4, ischemia + BH4 4.3 +/- 1.2 pmol/min per mg protein). Immunoblot after low-temperature sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the dimeric form of endothelial NOS decreased in the ischemic kidney and that it was restored by BH4. These results suggest that the decreased activity of endothelium-derived NO may worsen the ischemic tissue injury, in which depletion of BH4 may be involved. PMID- 10665938 TI - The effect of discharge voltage on renal injury and impairment caused by lithotripsy in the pig. AB - The present study was designed to determine the effects of shock wave voltage (kV) on lesion size and renal function induced by shock wave lithotripsy (SWL) in the 6- to 8-wk-old pig. Each SWL-treated pig received 2000 shock waves at 12, 18, or 24 kV to the lower pole calyx of one kidney. A group of sham SWL pigs served as time controls. Bilateral GFR, renal plasma flow (RPF), and para-aminohippurate (PAH) extraction were measured 1 h before and 1 and 4 h after SWL in all treated and sham animals. The kidneys were removed at the end of each experiment for morphometric analysis. The SWL-induced lesion increased significantly in size as shock wave energy was increased from 12 to 24 kV. PAH extraction, a measure of tubular function, was not significantly affected at 12 kV, was transiently reduced at 18 kV, and was reduced for the duration of the experiment at 24 kV. GFR and RPF, however, were significantly and similarly reduced at the 1 h post SWL period at all three kilovolt levels. At the 4-h post-SWL period, both GFR and RPF had returned to baseline levels. Lesion size and tubular injury were correlated with changes in kilovoltage, while changes in renal hemodynamics were already maximal at the lowest discharge voltage. These findings suggest that renal microvessels are highly sensitive to shock waves and that frank injury to tubules and vessels may be more closely related to discharge energy than is renal blood flow. PMID- 10665939 TI - Effect of gender on the progression of nondiabetic renal disease: a meta analysis. AB - There is previously published evidence that male gender is associated with a more rapid rate of progression of nondiabetic chronic renal disease. However, some investigators have concluded that no such association exists. To help resolve this issue, a meta-analysis was performed using 68 studies that met defined criteria and contained a total of 11,345 patients to evaluate the effect of gender on the progression of nondiabetic chronic renal disease. The results indicate that men with chronic renal disease of various etiologies show a more rapid decline in renal function with time than do women. PMID- 10665940 TI - Race is a major determinant of secondary hyperparathyroidism in uremic patients. AB - In the general population, blacks have higher parathyroid gland mass and circulating parathyroid hormone (PTH) levels than whites. This may predispose black patients to more severe parathyroid disease when renal failure develops. Therefore, racial differences in the severity of uremic hyperparathyroidism were examined in a population of patients with end-stage renal disease (ESRD). Among ESRD patients receiving hemodialysis or peritoneal dialysis, two or more values of intact PTH (immunoradiometric assay, pg/ml) obtained at least 90 d apart were available in 1270 prevalent cases (61.1% blacks, 51% males, and 31.1% diabetic), including 466 incident cases with onset of ESRD after 1993. Maximum PTH levels were analyzed as a function of race, gender, age, diabetic status, and levels of serum calcium, phosphorus, alkaline phosphatase, and aluminum. Using a stepwise multiple regression model, the determinants of maximum PTH in the order of their importance were black race, serum phosphorus, absence of diabetes, younger age, serum calcium, and female gender. The maximum PTH levels averaged 641.7 in blacks and 346.0 in whites after adjusting for age, gender, diabetic status, serum calcium, and phosphorus (P < 0.0001). In blacks compared with whites, the odds ratio (95% confidence interval) for adynamic bone disease (maximum PTH <150 pg/ml) was 0.26 (0.17 to 0.41), whereas the odds ratio for hyperparathyroid bone disease (mean PTH >500 pg/ml) was 4.4 (2.10 to 9.25). Race is a major independent determinant of uremic secondary hyperparathyroidism. Among ESRD patients, blacks may be at an increased risk for hyperparathyroid bone disease and whites for adynamic bone disease. PMID- 10665941 TI - Increasing the hematocrit has a beneficial effect on quality of life and is safe in selected hemodialysis patients. Spanish Cooperative Renal Patients Quality of Life Study Group of the Spanish Society of Nephrology. AB - Target hematocrit/hemoglobin values in dialysis patients are still controversial. The Spanish Cooperative Renal Patients Quality of Life Study Group (including 34 hemodialysis units) conducted a prospective, 6-mo study of the effect on patient functional status and quality of life of using epoetin to achieve normal hematocrit in hemodialysis patients with anemia. The possible adverse effects of increased hematocrit, patient hospitalization, and epoetin requirements were also studied. The study included 156 patients (age range, 18 to 65 yr). Given the minimal experience in the safety of increasing hematocrit in dialysis patients to normal levels with epoetin, stable patients on hemodialysis who had received epoetin treatment for at least 3 mo and had a stable hemoglobin level of > or = 9 g/dl were included in the study. Patients with antecedents of congestive cardiac failure, ischemic cardiopathy, diabetes mellitus, uncontrolled hypertension, cerebrovascular accident or seizures, malfunction of the vascular access or severe comorbidity (defined by a comorbidity index), and those over 65 yr of age were excluded from the study. Quality of life was measured with the Sickness Impact Profile (SIP) and Karnofsky scale. Patients completed questionnaires at home at onset and conclusion of the 6-mo study. Mean hematocrit increased from 30.9 to 38.4% and hemoglobin from 10.2 to 12.5 g/dl during the study. Health indicator scores improved significantly: mean Physical Dimension (SIP) from 5.38 to 4.1 (P < 0.005); mean Psychosocial Dimension from 9.2 to 7 (P < 0.001); mean global SIP from 8.9 to 7.25 (P < 0.001); mean Karnofsky scale score from 75.6 to 78.4 (P < 0.01). (SIP is scaled so that lower scores represent better functional status, and vice versa for the Karnofsky scale). Therefore, functional status and quality of life improved with increased hematocrit. No deaths occurred. Three patients (2%) were censored for hypertension and nine (5.7%) for thrombosis of the vascular access. The cumulative probability of thrombosis of the vascular access was 0.067. The average epoetin dose rose from 93 +/- 62 U/kg per wk at onset to 141 +/- 80 U/kg per wk at conclusion, a 51% increase. The number of patients hospitalized decreased and hospital lengths of stay were shorter during the study period than in the same patients in the 6-mo period preceding the study (P < 0.05). Nine patients (5.7%) had thrombosis of the vascular access. There were no changes in the prevalence of arterial hypertension, but three patients (2%) showed hypertension that was difficult to control. It is concluded that normalization of hematocrit in selected hemodialysis patients, i.e., nondiabetic patients without severe cardiovascular or cerebrovascular comorbidities, improves quality of life and decreases morbidity without significant adverse effects. PMID- 10665943 TI - Renal transplantation from non-heart beating donors: a promising alternative to enlarge the donor pool. AB - The aim of this study was to compare the survival and midterm function of kidneys from non-heart beating donors (NHBD) with those of kidneys from heart beating donors (HBD). From 1989 to 1998, 144 kidneys were procured from NHBD at the Hospital Clinico San Carlos in Madrid, of which 95 were transplanted. The kidney grafts were maintained from the moment of the diagnosis of cardiac arrest until the time of procurement by cardiopulmonary bypass. There was no significant difference in renal function and the number of rejection episodes between the NHBD and HBD transplants. The NHBD kidneys showed a 5.73-fold increase in the incidence of delayed graft function (adjusted relative risk 95% confidence interval, 2.82 to 11.62). One- and five-year survival rates for NHBD grafts were 84.6 and 82.7%, respectively, compared with 87.5 and 83.9% for HBD (P = 0.5767). Cox analysis showed that the predictive factors for worse NHBD graft survival were type of NHBD donor and the occurrence of corticoresistant rejection. Ninety of the NHBD organs were procured from subjects suffering irreversible cardiac arrest on the street who were transferred to our center for the sole purpose of donation. Fifty-four of these kidneys were transplanted and all showed primary function. When a strict protocol is adhered to, the outcome of renal transplant from NHBD compares well with that from HBD. It is believed that the high number of organs obtained from subjects undergoing irreversible cardiac arrest on the street might encourage the adoption of new criteria for the management of this type of pathology with the ultimate goal of kidney donation. PMID- 10665942 TI - Impact of cyclosporin A pharmacokinetics on the presence of side effects in pediatric renal transplantation. AB - Cyclosporin A (CsA) is a potent immunosuppressant that has many side effects, including hypertrichosis, gingival hyperplasia, and tremor. To evaluate whether there is a relationship between the CsA-pharmacokinetics (PK) and these side effects, their presence and intensity were observed in 46 renal transplanted children/adolescents during two regular visits, and the occurrence of the side effects was correlated with CsA-PK. CsA doses had been unchanged for at least 6 mo. CsA blood concentrations were measured at time 0, and 1, 2, and 4 h after the CsA morning dose. An abbreviated area under the curve (AUC) was calculated using C0, C2, and C4. Hypertrichosis positively correlated with C2, C4, Cmax, and AUC. An AUC > or = 4158 ng/ml per h was the best predictor for the presence of hypertrichosis. Tremor was also positively correlated with C2, Cmax, and AUC. A Cmax > or = 878 ng/ml was the best predictor for the appearance of tremor. These values of Cmax and AUC are within the therapeutic range of CsA as demonstrated by the studies of calcineurin inhibition by CsA. Gingival hyperplasia was not associated with any of the CsA-PK studied parameters. However, it was associated with the concomitant use of nifedipine. These data show that there is a correlation between the CsA side effects and its pharmacokinetics and that it is possible to decrease the incidence and intensity of such side effects by monitoring the CsA-PK parameters, providing they are under or at the proposed cutoff levels. Nifedipine should also be avoided to reduce the presence of gingival hyperplasia. PMID- 10665944 TI - Glycosaminoglycans: use in treatment of diabetic nephropathy. PMID- 10665945 TI - Metabolic alkalosis. PMID- 10665946 TI - Vasopressin: induced structural change in toad bladder luminal membrane. 1975. PMID- 10665947 TI - Disease incidence in dairy herds in the southern highlands district of New South Wales, Australia. AB - The purpose of this study was to determine clinical disease incidence in eight non-seasonally calving, pasture-fed dairy herds in the southern highlands district of New South Wales. This was a longitudinal population study. The study included all cows that calved between January 1994 and December 1995 and consisted of 2111 lactation records from 1430 cows. The incidence of the more common diseases were: calving-associated disorders, 18.0 cases per 100 calvings (95% CI 16.4-19.8 cases per 100 calvings); metabolic disorders, 5.5 cases per 100 cow-yr at risk (95% CI 4.5-6.6 cases per 100 cow-yr at risk); reproductive-tract disorders, 22.3 cases per 100 cow-yr at risk (95% CI 19.2-25.8 cases per 100 cow yr at risk); udder disorders, 17.6 cases per 100 cow-yr at risk (95% CI 15.9-19.5 cow-yr at risk) and lameness, 3.7 cases per 100 cow-yr at risk (95% CI 2.9-4.7 cow-yr at risk). In agreement with dairy-cow disease-incidence studies conducted elsewhere, disorders of the reproductive-tract and udder were the most frequent clinical conditions encountered. These findings confirm that dairy herd-health programs should emphasise the control of these two groups of disorders. PMID- 10665948 TI - Temporal patterns of domestic and wildlife rabies in central Namibia stock ranching area, 1986-1996. AB - Eleven years (1986-1996) of wildlife- and domestic-rabies data from the agriculture stock-ranching area of central Namibia were studied using time-series analysis. Nine hundred and sixty three rabies cases were observed in domestic ruminants (5.4 cases/mo), black-backed jackals (Canis mesomelas, 1.3 cases/mo), domestic dogs (0.5 case/mo), and bat-eared foxes (Otocyon megalotis, 0.1 case/mo). The incidence of rabies for all species did not change significantly over the whole study period. However, seasonal variations with an increase in the number of cases between June and November of each year, as well as 34 yr cyclical fluctuations were identified in domestic ruminants and black-backed jackals. The black-backed jackal time-series variable was a significant predictor of the domestic-ruminant and dog time-series variables. The rainfall seasonality combined with the seasonal reproductive pattern of the black-backed jackal appeared to be plausible explanations for the seasonal variations of rabies. However, there was no overall significant correlation between the cyclical weather fluctuations and the 3-4 yr cyclical rabies variations. PMID- 10665949 TI - A stochastic partial-budget analysis of an experimental Pasteurella haemolytica feedlot vaccine trial. AB - A field trial compared a modified Pasteurella haemolytica biotype A serotype 1 leukotoxin vaccine to a commercial vaccine during March-July 1995 in a Natal Midlands, South African, feedlot. Weaners/long weaners purchased by the feedlot were allocated systematically into test vaccine and control vaccine groups of 1241 and 1240 head, respectively, and fed in groups of approximately 200 head. Morbidity and mortality were monitored until the animals were marketed. Details of pleuritis and pneumonia at veterinary meat inspection were recorded for 409 test-vaccinated and 424 control-vaccinated cattle. An increase in morbidity but not mortality risk of respiratory disease was shown between test (13.8% morbidity) and control (11.4% morbidity) groups. Cattle with a processing weight <245 kg were 1.4 times more likely to develop respiratory diseases than cattle with a processing weight > or =245 kg. Cattle bought on auction were 1.6 times more likely to develop respiratory disease than cattle bought at private sales. A partial farm budget incorporating Latin Hypercube sampling of uncertain variables was done to obtain the distribution of possible financial outcomes if the test vaccine were used. Impact (sensitivity) analyses indicated that median weight of carcass cut away had the greatest impact on the profit margin. The partial farm budget highlighted the importance of reducing sub-clinical lesions in a feedlot. PMID- 10665950 TI - Soil type as a putative risk factor of ovine and caprine paratuberculosis seropositivity in Spain. AB - Relationships between soil type and ovine and caprine paratuberculosis in the Avila region (central Spain) were evaluated using data from a cross-sectional study of the most-important diseases of small ruminants in this Spanish region between 1996 and 1997. Questionnaire data from 61 herds (38 ovine and 23 caprine) and 1451 serum samples (1041 ovine and 410 caprine) were used. Herd paratuberculosis (herds were scored as positive to paratuberculosis if any of the serum samples was positive in an agar-gel immunodifussion) was the outcome of interest, whereas soil type in the municipality where farms were located was the predictor variable. Other variables related to soil and soil usage, and herd size, replacement, main food production and animal species were also introduced into the multivariable logistic regression. The final model contained only two independent variables: the predictor variable soil type (coded as two dummy variables ST-1 and ST-2) and herd size (dichotomized at the highest quartile). The estimated Odds Ratios were 25.9 (95% CI: 1.6, 411) for ST-1 (entisols as soil type) and 3.5 (95% CI: 0.3, 45) for ST-2 (inceptisols as soil type). PMID- 10665951 TI - Failure to identify non-bovine reservoirs of Mycobacterium bovis in a region with a history of infected dairy-cattle herds. AB - The State of Texas had the most (cumulative) tuberculous cattle herds of any state in the United States during the decade ending in 1997. Of the cumulative 18 infected herds in Texas, 12 herds were concentrated in El Paso County (designated the 'El Paso milkshed'). To identify whether non-bovine reservoirs were a source of Mycobacterium bovis infection of cattle in this region, an investigation was conducted on the premises of 14 dairy herds (12 tuberculous and 2 non-affected herds) between May 1995 and June 1997. None of the 670 mammalian, avian and environmental (soil, water and air) samples collected and cultured from the premises of these herds was positive for the presence of M. bovis. None of the 119 human urine samples obtained from employees of these dairies was culture positive for M. bovis. Of 124 dairy-farm workers with tuberculin skin-test results, 48 showed positive test results. There was, however, no difference in percentages of positive skin-test results between farms without, and farms having, bovine tuberculosis within the last two years or longer. The percentage of positive reactions did not increase with length of time employed at a dairy with a history of confirmed tuberculosis. These findings suggest that non-bovine reservoirs appear not to be a factor responsible for tuberculosis of cattle in the El Paso milkshed. PMID- 10665952 TI - Effect of ethidium bromide intercalation on DNA radiosensitivity. AB - PURPOSE: To assess the influence of the intercalating drug ethidium bromide (EtBr) on the yields of single strand breaks (ssb) induced by fast neutrons in supercoiled pBR322 plasmid and in a linear DNA restriction fragment. MATERIALS AND METHODS: The yield of ssb in the plasmid was measured by agarose gel electrophoresis. The proportion of fragments bearing one ssb and the probability of breakage at each nucleotide site was determined using sequencing gel electrophoresis. The volume variations due to the intercalation of EtBr were calculated. The expected radio-modifying effect at each nucleotide site of the linear fragment was evaluated using a reported simulation procedure. RESULTS: The ssb yield in the plasmid increased for concentrations up to 0.04 drug/bp and fell back in the range 0.04-0.1 drug/bp. For the linear DNA, only a slight protective effect was observed over the whole concentration range. The effect was almost the same at all nucleotide sites. CONCLUSION: For the linear DNA fragment, radioprotection was mainly due to scavenging of OH* radicals by the intercalated drug. For the plasmid, the radio-modifying effect results mainly from the variation of its effective volume, due to the modification of superhelicity. PMID- 10665953 TI - Effects of low dose irradiation on TK6 and U937 cells: induction of p53 and its role in cell-cycle delay and the adaptive response. AB - PURPOSE: To investigate the effect of small doses of radiation on the cell-cycle and related processes, and to determine the capacity of small doses of radiation to induce an adaptive response. MATERIALS AND METHODS: TK6, a lymphoblast cell line with wild-type p53, and U937, a monocytic leukaemia cell line with mutant, inactive, p53 were exposed to gamma ray doses ranging from 0.1 Gy to 3 Gy. Cell cycle distributions and cyclin B1 were assessed by flow cytometry, and p53 and p21 protein levels were measured by Western blotting. Apoptosis was determined by fluorescence microscopy after staining with Hoechst 33342, and by measurement of the pre-G1 cell population by flow cytometry. Micronuclei were determined in cytokinesis-blocked cells by fluorescence microscopy. RESULTS: In TK6 cells, radiation exposure induced elevated p53 and p21 levels and delayed expression of cyclin B1. No changes in these parameters were found in U937 cells. Although both cell lines arrested in G2/M after larger doses of radiation, G2/M-arrest occurred after 0.1 Gy and 0.3 Gy in TK6 cells only. An apoptotic adaptive response was induced in both cell lines by a 0.1 Gy priming dose but an adaptive response with respect to micronuclei was observed only in U937 cells. CONCLUSIONS: The radiation adaptive response can occur in the absence of wild-type p53. A small dose of radiation may not protect cells against both apoptosis and cytogenetic damage caused by a subsequent larger dose of radiation. PMID- 10665954 TI - Production of chromatid breaks by single dsb: evidence supporting the signal model. AB - PURPOSE: The signal model proposes that all chromatid breaks arise from a single DNA double strand break (dsb) via a recombinational exchange mechanism. Here the prediction that chromatid breaks arise from a single dsb is tested. METHOD: The genetically engineered Chinese hamster cell line GS19-43 containing a unique yeast I-SceI recognition site was treated with I-SceI endonuclease (Meganuclease) in the presence of the porating agent streptolysin O. Chromatid breaks were scored at 4h, chromosome breaks at 18 and 22h following treatment (cells used for a 4h fixation were prelabelled with BrdU over two cell-cycles). Positive controls were treated with the restriction endonuclease Pst 1. RESULTS: I-SceI endonuclease produced chromatid breaks and at higher enzyme concentrations isochromatid breaks but no chromatid interchanges. About 16% of the chromatid breaks had a 'colour-switch' between the sister-chromatids at the site of breakage, as revealed by FPG staining. At the longer fixation times (18 and 22 h) chromosome breaks were observed, but again no interchanges were seen. Chromatid and chromosome breaks always appeared on the same chromosome. CONCLUSIONS: The production of chromatid breaks from a single dsb fulfils the prediction of the signal model. Moreover, the production of colour-switch breaks at a similar frequency to that for ionizing radiation indicates that chromatid breaks are produced via recombinational exchanges, a significant proportion of which occurs between sister chromatids. The majority is intrachromatid, not involving strand switches. The absence of interchromosomal exchanges at all fixation times indicates a requirement of two dsb in two different chromosomes for their formation. PMID- 10665955 TI - Complex chromosome aberrations in peripheral blood lymphocytes as a potential biomarker of exposure to high-LET alpha-particles. AB - PURPOSE: To determine the induction and transmission, to second and third division cells, of complex chromosome aberrations in peripheral blood lymphocytes after exposure to high-LET alpha-particles in vitro. MATERIALS AND METHODS: Separated peripheral blood lymphocytes collected from four healthy donors were irradiated in vitro with either high-LET alpha-particles (121 keV/microm; 0.5 Gy) or low-LET X-rays (250kV constant potential; 3 Gy). Cells were harvested in first, second and third division post-irradiation and chromosome aberrations observed at each cell division were analysed by combining the techniques of FISH and DAPI/Hoechst 33258 harlequin staining. Whole chromosome probes were used for chromosomes 1, 2 and 5, together with a pan-centromeric probe and the resulting chromosome 'painting' patterns were classified according to the Savage and Simpson (S & S) scheme (Savage and Simpson 1994a, Savage and Tucker 1996). RESULTS: A greater proportion of complex chromosome aberrations was observed, defined as involving three or more breaks in two or more chromosomes, relative to total exchanges, after exposure to 0.5 Gy alpha-particles (mean 1 track/cell) than after the high reference dose of 3 Gy X-rays (49-56% and 20-22%, respectively). Qualitatively, alpha-particles induced chromosome aberrations of far greater complexity than those observed after X-rays. This was reflected by both the rapid reduction in the percentage of damaged cells between first and second division indicative of cell death, and the spectrum of aberration types observed in second and third division cells post-irradiation. Regardless of this complexity, 15% of the complexes induced by alpha-particles at first division were potentially transmissible and by third division, transmissible-type complexes, specifically insertions, represented the predominant complex type (65%). CONCLUSION: Transmissible-type complexes were observed, specifically insertions, in both second and third division cells after exposure to high-LET alpha-particles (0.5 Gy) in vitro. The authors predict these cells to be stable and to be capable of persisting through many cell generations. Considering that the induction of complex chromosome aberrations by low-LET radiation is strongly dependent on dose, so that they are expected to be undetectable at environmental exposures, insertions are much more likely to be a characteristic feature of high LET radiation at all doses. From this the usefulness of insertions as biomarkers of past exposure to environmentally relevant doses of high-LET alpha-particles is supported. PMID- 10665956 TI - Histone-like protein HU is required for recA gene-dependent DNA repair and SOS induction pathways in UV-irradiated Escherichia coli. AB - PURPOSE: Escherichia coli HU protein exists as a heterodimer composed of two highly homologous subunits, HU-1 and HU-2, encoded by the hupB and hupA genes, respectively. It introduces negative supercoils into a relaxed circular DNA. Various roles of HU have been suggested in cellular processes such as DNA replication and transcription. The present experiments were designed to understand the role of HU in DNA repair processes in E. coli. MATERIALS AND METHODS: The sensitivity of hupA/hupB mutants of E. coli to the lethal and mutagenic effects of UV was compared with that of a wild-type strain. The effect of the hupAhupB mutations in SOS induction was also examined. RESULTS: The hupAhupB mutations increased the UV sensitivity of E. coli. Nucleotide excision repair was unaffected by the deficiency of HU. On the other hand, E. coli hupAhupB mutants were sensitive to UV in the recA+recB+recF background but not in the recArecB+recF+ or recA+recBrecF+ background. The frequency of UV-induced mutation to rifampicin resistance was significantly reduced in the hupAhupB mutants, and the induction of the recA::lacZ and umuC::lacZ fusion genes was also suppressed in the mutants. CONCLUSIONS: HU protein plays a critical role in the recA, recB-dependent recombinational DNA repair and SOS induction pathways in UV irradiated E. coli. PMID- 10665957 TI - Effects of radical scavengers on radiation-induced DNA double strand breaks. AB - PURPOSE: To investigate possible effects of Tris and phenol on the dynamic properties of gamma-irradiated DNA molecules in addition to their well known scavenging capacity. MATERIALS AND METHODS: Native and fragmented calf thymus DNA molecules were exposed to various doses of 60Co gamma-rays at approximately 4.5Gy/min. Using thermal transition spectrophotometry, pulsed field gel electrophoresis and standard agarose gel electrophoresis, the effects of Tris, phenol and NaCl on the double helix to single coil thermal transition temperature, Tm, and the yield of the double-strand breaks (Gdsb) of the irradiated DNA molecules have been studied. RESULTS: DNA molecules exposed to gamma-rays showed a decreased Tm and a corresponding increase of the Gdsb yield. Tris, as well as phenol, exhibited a strong protection against preventing these radiation-induced alterations. In addition, both substances strongly affected the thermal stability of the non-irradiated DNA samples. These results, compared with data obtained by NaCl and its effects on DNA thermostability and Gdsb, revealed that in the presence of both scavengers the observed dsb decrease was correlated to an increased molecular stability of DNA. CONCLUSIONS: This work suggests that the total protective effect of Tris and phenol against radiation-induced dsb is mainly attributed to their well-known radical scavenging properties, while relatively minor protective effects arise from their contribution to an increased molecular stability of DNA. PMID- 10665958 TI - Technical report: SYBR Green I and the improved sensitivity of the single-cell electrophoresis assay. AB - The single-cell electrophoresis (comet) assay is an established method for measuring radiation-induced strand breaks in DNA. The extent of damage is quantified in individual cells by assessing the intensity and distribution of a fluorescent DNA-binding dye using image analysis software. Using the Kinetic Imaging Komet3 system, it is demonstrated that the fluorochrome SYBR Green I improves the resolution and sensitivity of the comet assay, particularly for measuring radiation-induced double strand breaks. PMID- 10665959 TI - Comparison of biological effects of DNA damage induced by ionizing radiation and hydrogen peroxide in CHO cells. AB - PURPOSE: Free OH radicals are considered to be the common mediator of DNA damage after ionizing radiation and oxidative stress. In particular, double-strand breaks (dsb) have a major impact on cell killing after irradiation, while the mechanism of cell killing is less clear for oxidative injury. The latter not only affects DNA, but also equally other cell compartments, such as membranes and mitochondria, which may trigger cell death. This study intended to clarify the relationship between DNA damage induction, repair and cell inactivation for hydrogen peroxide and ionizing radiation. MATERIALS AND METHODS: Chinese hamster ovary (CHO) cells were treated with H2O2 in serum-free medium in combination with/ without X-irradiation. DNA damage was measured using the alkaline unwinding method or neutral constant-field gel electrophoresis. Cell survival was recorded using the colony-formation assay. RESULTS: Hydrogen peroxide induced a large number of single-strand breaks (ssb>36000/cell) without impairing cell survival. This number reached a maximum (36 Gy-equiv. at 3 x 10(-4) mol/dm3) without further increase after higher concentrations. Repair kinetics of ssb were similar to those after irradiation. Dsb were found only after very high concentrations of H2O2 (>3 x 10(-2) mol/dm3), which is different from irradiation which generated ssb and dsb in the same dose range. A linear-quadratic increase of dsb was found with increasing concentrations of H2O2 suggesting a single or a pairwise action of OH radicals to form a dsb. After either irradiation or peroxide treatment cell killing was observed only after doses which also allowed dsb detection. The number of dsb calculated per lethal event was in the same range but slightly higher after irradiation (1.7-fold) than after H2O2 treatment. CONCLUSIONS: Cell killing after irradiation or hydrogen peroxide appears to be due to dsb, whereas cells withstand large numbers of single-strand lesions and other types of non-DNA damage occurring at lower concentrations of hydrogen peroxide. The number of ssb saturates at intermediate concentrations of H2O2 suggesting that a limited amount of chromatin-bound metal ions is available for OH radical generation. PMID- 10665960 TI - Modification of murine adult haemopoiesis and response to methyl nitrosourea following exposure to radiation at different developmental stages. AB - PURPOSE: To investigate the hypothesis that the developmental phase at which an individual encounters radiation damage affects its long-term sensitivity to a subsequent tumourigenic insult. MATERIALS AND METHODS: Either the pregnant C57B16 mouse was exposed to 137Cs gamma-rays at 4 or 15 days post-conception (embryonic and foetal stages respectively) or BDF1 offspring were irradiated at 4 or 21 days of age (neonatal and juvenile stages). Offspring were either assayed for changes in bone marrow stem cells and committed progenitors at 6, 12 and 18 weeks of age, or injected with the chemical carcinogen methyl nitrosourea (MNU) at 10 weeks of age and monitored for onset of neoplasia. RESULTS: Gamma-irradiation induced a persistent long-term deficit in stem cells in all irradiated animals, with the foetal stage appearing most radiosensitive. However, femoral cellularity, committed progenitor cell numbers and peripheral blood counts were unaffected. When offspring were exposed to MNU, the incidence of malignancy was significantly enhanced in animals irradiated at the foetal, neonatal and juvenile stages. CONCLUSIONS: This study has shown that exposure to ionizing radiation at the foetal, neonate or juvenile stages of development induces residual haemopoietic damage and increases oncogenic susceptibility to a subsequent exposure to MNU. PMID- 10665961 TI - Transformation by radiation of rat foetal glial cells. AB - PURPOSE: To establish and characterize an in vitro model of radiation-induced transformation of normal glial cells. MATERIALS AND METHODS: During the last week of gestation, pregnant Sprague-Dawley rats were either irradiated at 3.5 Gy (0.022 Gy h(-1)) with a 60Co source or sham irradiated. On day 21 of gestation, cortical nerve cells from foetuses were isolated, and then maintained in culture for about 100 passages, in presence of 10(-9) g/ml of tetradecanoyl phorbol acetate (TPA). To follow transformation, various parameters: cell type, proliferation, clonogenicity, karyotypes and tumorigenicity, were studied at different passages. RESULTS: As the number of passages increased, control cells lost their glial morphology and were immortalized. They kept on expressing specific markers of type 2 astrocytes (glial fibrillary acid protein (GFAP) and A2B5). Karyotypes remained near diploid. At all passages tested, they were not tumorigenic in nude mice. Irradiated cells expressed the 2A progenitor cell specific markers: GFAP, vimentin and A2B5. Karyotypes evolved toward polyploidy and cells displayed an iso 7 and a marker. These changes were synchronous with modifications in tumorigenicity. Metastases were even observed in nude mice. CONCLUSIONS: Cells from irradiated animals were fully transformed, while cells from sham irradiated animals were only immortalized. PMID- 10665962 TI - Induction of solid tumours in the Swiss albino mouse by low-dose foetal irradiation. AB - PURPOSE: To study the tumourigenic effect of prenatal low-dose gamma-irradiation in the mouse. METHODS AND MATERIALS: Pregnant Swiss albino mice were exposed to 0.1-1.5 Gy gamma-radiation on days 14 or 17 of gestation. The F1 offspring were observed up to 18 months of age. All the mice were killed at 18 months and the incidence of tumours in different organs was recorded. RESULTS: Exposure to doses from 0.1 to 1.5 Gy on days 14 or 17 of gestation produced a linear-quadratic dose dependent increase in tumour incidence in adult F1 mice. The main organs affected were the ovary, uterus, liver and spleen. The highest incidence was observed in the ovaries, which was significantly higher than spontaneous incidence, even at 0.25 Gy. In other organs the tumour incidence was not significant compared with controls at doses < 0.5-1.0 Gy. Tumours in the ovary and uterus developed at an earlier age than in the liver and spleen. CONCLUSIONS: Exposure to gamma radiation < 1.0 Gy at the foetal period (days 14 or 17 of gestation) can cause induction of tumours in the Swiss albino mouse. The carcinogenic effect, particularly on the ovary among the female mouse, is detectable after low-dose foetal irradiation. PMID- 10665963 TI - Microdistribution and localized dosimetry of the alpha-emitting radionuclides 239Pu, 241Am and 233U in mouse femoral shaft. AB - PURPOSE: To analyse the temporal change in microdistribution of 239Pu, 241Am and 233U in mouse femur and to compare the calculated radiation doses with regions of the bone marrow thought to contain target cells for osteosarcoma and leukaemia with relative risk for those diseases. MATERIALS AND METHODS: Neutron-induced and alpha-track autoradiographs were prepared from femora of the CBA/H mouse that had been injected with 40 kBq kg(-1) radionuclide between 1 and 448 days previously. Computer-based image analysis of the autoradiographs was performed and dosimetric methods applied to obtain radiation dose-rates to different regions of the marrow cavity. RESULTS: Initially each radionuclide deposited on endosteal and periosteal bone surfaces; 241Am was additionally deposited on vascular canal surfaces. Redistribution resulted in 233U being incorporated into bone, while 239Pu and 241Am showed transfer into both bone volume and marrow. Accumulation in the central marrow peaked at 112-224 days post-injection, but subsequently was cleared by 448 days. Cumulative doses to both osteosarcomagenic and myeloid leukaemogenic target cell regions showed the trend 239Pu > 241Am > 233U. CONCLUSIONS: Calculation of cumulative doses to a 10-microm layer of marrow adjacent to bone surfaces appears to be a suitable predictor for risk of osteosarcoma. Risks of myeloid leukaemia in the mouse are better predicted by considering the central marrow as the target region rather than average dose to all marrow. PMID- 10665964 TI - Efficacy of 3,4,3-LIHOPO for reducing neptunium retention in the rat after simulated wound contamination. AB - PURPOSE: The ligand 3,4,3-Li(1,2-HOPO) was tested for Np removal after intramuscular injection of 237Np nitrate in rats. MATERIALS AND METHODS: Two experiments were performed, one with simultaneous injection of neptunium and LIHOPO at dosages ranging from 3 to 200 micromol kg(-1) and the other with delayed administration of LIHOPO 30 micromol kg(-1) from 5 min to 30 min after Np injection. RESULTS: The data obtained after simultaneous injections showed that the ligand dosage effectiveness was not linear and depended on the tissues being considered. For bones, the best results were obtained with 200 micromol kg(-1) LIHOPO, where retention was reduced to 11% of controls. Maximum efficacies for removal in liver and kidney were obtained with 30 micromol kg(-1) LIHOPO, where retention was reduced to 39% and 1.6% of controls, respectively. At higher dosages, LIHOPO seemed to have a reverse effect on these tissues, demonstrated by a significant accumulation of the radionuclide. The delayed administration of LIHOPO dramatically decreased its efficacy. When administered 5 min after Np, LIHOPO was still efficient (60%, 37%, 7% of controls in bone, liver, kidneys, respectively) but not when treatment was delayed to 30 min. CONCLUSIONS: These results demonstrated that LIHOPO was able to complex Np at the wound site but not after translocation to blood. PMID- 10665965 TI - Commentary on the Second Event Theory of Busby. PMID- 10665966 TI - Commentary on the Second Event Theory of Busby by A. A. Edwards and R. Cox. PMID- 10665967 TI - Think metabolically. PMID- 10665968 TI - Persisting motor control problems in 11- to 12-year-old boys previously diagnosed with deficits in attention, motor control and perception (DAMP). AB - The aim of this study was to examine whether boys who had been previously diagnosed between the ages of 5 and 8 years with deficits in attention, motor control and perception (DAMP) still have problems with motor control, which influence their spare-time and everyday activities, at 11 to 12 years. The study comprised a well defined cohort of 10 boys with DAMP and a control group of 20 boys without DAMP matched for age, height, and weight. The Movement Assessment Battery for Children was used to assess motor control in ability to perform everyday activities, and the spare-time activities in which the boys participated were recorded. Individually, the boys previously diagnosed with DAMP had a markedly higher total score (poor performance) than the boys without DAMP (P<0.001). The everyday activities of boys with DAMP were significantly affected, and they chose to participate in different sports from the control boys, i.e. none participated in team sports. The present study does not support the concept of improvements in motor control with age in children with DAMP. PMID- 10665969 TI - Corpus callosum may be similar in children with ADHD and siblings of children with ADHD. AB - No previous studies have used morphological neuroimaging to compare children with ADHD with siblings of children with ADHD. To test the hypothesis that the total size of the corpus callosum is altered in children with hyperkinetic disorder, the corpus callosum was outlined from a single midline protondensity weighted slice (containing the septum pellucidum). Fifteen boys with a refined phenotype of ADHD (mean age 10.2 years) and 15 healthy male siblings of children with ADHD (mean age 10.6 years) were enrolled in the study. The two groups were compared for global brain size and the callosal areas of Witelson. No significant differences were found between the study and comparison groups for any of the corpus callosum areas, even after age, global brain size, and handedness were covaried (using MANOVA). In addition, corpus callosum sizes do not seem to differ between children with ADHD and unaffected siblings of children with ADHD. Clinicians should not base their pathophysiological diagnosis of this disorder on an abnormality of callosal development. PMID- 10665970 TI - Fetal growth and subsequent mental health problems in children aged 4 to 13 years. AB - To test the hypothesis that children with suboptimal fetal growth have significantly poorer mental health outcomes than those with optimal growth, a population random sample survey of children aged 4 to 16 years in Western Australia in 1993 was conducted. The Child Behavior Checklist (Achenbach 1991a) and the Teacher Report Form (Achenbach 1991b) were used to define mental health morbidity. Survey data for 1775 children aged 4 to 13 years were available for linkage with original birth information. The percentage of expected birthweight (PEBW) was used as the measure of fetal growth. Children below the 2nd centile of PEBW who had achieved only 57% to 72% of their expected birthweight given their gestation at delivery were at significant risk of a mental health morbidity (OR 2.9, 95% CI 1.18, 7.12). In addition, they were more likely to be rated as academically impaired (OR 6.0, 95% CI 2.25, 16.06) and to have poor general health (OR 5.1, 95% CI 1.69, 15.52). PMID- 10665971 TI - Parental and professional agreement in developmental assessment of very-low birthweight and term infants. AB - The aim of this prospective follow-up study was to evaluate the accuracy of a parent-completed questionnaire compared with professionally detected developmental delay. Parents of 108 very-low-birthweight (VLBW) infants and parents of 279 term control infants completed the German version of the Revised Prescreening Developmental Questionnaire (R-PDQ) at the corrected age of 12 months. Simultaneously, infants underwent developmental examination using the Griffiths Developmental Scale. Sixty-nine VLBW infants were classified as not delayed, 16 as delayed by both methods (conegativity 76% and copositivity 94%), as compared to 240 and six term control infants (conegativity 88%, copositivity 94%). The questionnaire suggested delay in 22 VLBW infants and 32 control infants, which was not substantiated by professional examination (P=0.006). In contrast, examination-diagnosed delay was missed by the questionnaire in one infant in each group. The R-PDQ is a reliable monitoring instrument for both VLBW and term infants at the age of 12 months. Parents of VLBW infants tend to underestimate their infants' development. PMID- 10665972 TI - Predictive utility of the Bayley Infant Neurodevelopmental Screener (BINS) risk status classifications: clinical interpretation and application. AB - Predictive validity and clinical implications of the increasingly popular Bayley Infant Neurodevelopmental Screener (BINS) risk status classifications have not been previously reported. In this longitudinal follow-up study, the BINS was administered to high-risk infants at 6, 12, and 24 months of age, and the McCarthy Scales at 3 years of age. Ninety-two children were evaluated at 6 and 36 months, 105 at 12 and 36 months, and 118 at 24 and 36 months; 190, 125, and 140 infants were included in the comparisons at 6 to 12, 6 to 24, and 12 to 24 months. BINS risk status was classified as low, moderate, or high; or as a binary variable, LOWRISK/HIGHRISK. The three BINS items groups were moderately correlated. Consistency was most variable in the moderate-risk group. BINS risk was predictive of 36-month function in 18 out of 18 comparisons. Odds ratios, ranging from 2.76 to 54.70, were significant in 15 out of 18 logistic models. An early high-risk classification was associated with increased probability of later developmental morbidity. The BINS offers an alternative to detailed assessment in high-volume clinical applications and has good concurrent and predictive validity. PMID- 10665973 TI - Biomechanical transformation of the gastroc-soleus muscle with botulinum toxin A in children with cerebral palsy. AB - Objective measures (kinematics and kinetics) were used to study prospectively the effects of botulinum toxin A (BTX/A) on the gastro-soleus muscle in ambulant children with cerebral palsy. In this prospective before and after trial, 15 children with diplegia and 10 children with hemiplegia were studied (mean age 5 years 7 months, range 4 years to 9 years). A range of standardized clinical measures was undertaken but the emphasis for this report is on the three dimensional gait analysis (3DGA) results. All children showed improvements in sagittal ankle kinematics, as has been previously reported. Two new measures of ankle kinetics were devised: ankle moment quotient (AMQ), and ankle power quotient (APQ). Before intervention, ankle moments were characterized by a 'double bump' ankle moment. A typical abnormal baseline ankle-power curve was triphasic with an initial trough of absorption followed by abnormal mid-stance power generation, instead of the usual A1 pattern, and reduced terminal stance power generation (A2). Three weeks after treatment with BTX/A alone there was a statistically significant improvement of AMQ and APQ; some patients required potentiation of BTX/A with a short period of serial casts. Both groups (BTX/A alone and BTX/A plus casting) continued to show improvement in ankle kinetics from baseline after 12 and 24 weeks. This is the first study to demonstrate improvements in the typical abnormal ankle kinetics which we believe provides evidence of the 'biomechanical transformation of muscle'. PMID- 10665974 TI - Ankle spasticity and strength in children with spastic diplegic cerebral palsy. AB - Ankle spasticity and strength in 27 children with spastic diplegic cerebral palsy (CP) (mean age 9 years, range 3 to 18 years) and a group of 12 children without CP (comparison group) (mean age 9 years, range 5 to 18 years) were observed. To measure spasticity, a KinCom dynamometer dorsiflexed the passive ankle at five different speeds and recorded the resistive plantarflexion torques. Work values for the torque-angle data were calculated at each speed. Using this data, linear regression was used to measure spasticity. To measure strength, the dynamometer rotated the ankle from maximum dorsiflexion to maximum plantarflexion at a speed of 10 degrees/s while the child performed a maximum plantarflexion concentric contraction. The movement was reversed to record maximum dorsiflexion. Maximum torques and work by the plantarflexors and dorsiflexors were calculated. The group with CP had significantly more spasticity in the plantarflexors and significantly less strength in the plantarflexors and dorsiflexors than the group without CP. Results provide objective information quantifying ankle spasticity and strength in children with CP. PMID- 10665975 TI - A study of EEG, electroretinogram, visual evoked potential, and eye movements in classical lissencephaly. AB - EEG, flash electroretinogram (ERG), and visual evoked potential (VEP) findings are described in eight children with classical lissencephaly (six girls, two boys), with a mean age of 17.6 months (range 2 to 60 months). The EEG shows typically high-voltage activity. Eye movements were formally recorded in two patients, and both showed features associated with oculomotor apraxia. The ERG and VEP to flash stimulation were normal in all cases. Two subjects had pattern reversal stimulation, and their pattern VEPs were within normal limits. Some patients with lissencephaly may appear to have delayed visual maturation on first presentation, and EEG and eye movement studies are valuable in indicating neurological deficiency at an early stage in these subjects. PMID- 10665976 TI - Severe bronchopulmonary dysplasia increases risk for later neurological and motor sequelae in preterm survivors. AB - Preterm children who develop severe chronic lung disease may be developmentally compromised by exposure to hypoxic episodes. This study aims to determine if children with severe bronchopulmonary dysplasia (BPD) who required home oxygen therapy were at greater risk for neurological and motor deficits at school age than preterm peers without BPD. This study evaluated 27 subjects with BPD and 27 preterm control infants matched for gestational age, birthweight, sex, and year of birth at a mean age of 9.9 years (2.0 SD) using standardized neuromotor outcome measures. Pair-matched comparisons and regression analyses were used to determine if subjects with BPD were at increased risk for neuromotor sequelae. Neurological abnormalities, including subtle neurological signs, cerebral palsy, microcephaly, and behavioral difficulties were highly prevalent in the BPD group (71% compared with 19% in control group, P<0.005). Over half the BPD cohort had difficulties in gross and/or fine motor skills. There were significant differences in postural stability between groups. Duration of hospitalization and home oxygen treatment, and decreased lung function at school age, markers of severity of illness, correlated with motor outcomes. The findings underline the importance of preventing the cardiorespiratory complications associated with chronic lung disease to minimize disability in preterm children. For children with severe BPD, better recognition and subsequent remediation of neuromotor impairments that manifest at school age may help maximize their functional potential. PMID- 10665977 TI - New ocular movement detector system as a communication tool in ventilator assisted Werdnig-Hoffmann disease. AB - A non-contact communication system was developed for a ventilator-assisted patient with Werdnig-Hoffmann disease who had lost all voluntary movements except for those of the eye. The system detects the extraocular movements and converts them to either a 'yes' signal (produced by one lateral eyeball movement) or a 'no' signal (produced by two successive lateral eyeball movements) using a video camera placed outside the patient's visual field. The patient is thus able to concentrate on performing a task without any intrusion from the detection system. Once the setting conditions of the device have been selected, there is no need for any resetting, as the patient is unable to move his body. In addition to playing television games, the child can use the device to select television channels, compose music, and learn written Japanese and Chinese characters. This seems to broaden the patient's daily world and promote mental development. PMID- 10665978 TI - Posture and motility in preterm infants. PMID- 10665979 TI - Linking motor impairment to function. PMID- 10665980 TI - Current status of artemisinin and its derivatives as antimalarial drugs. AB - Artemisinin is a promising and a potent antimalarial drug, which meets the dual challenge posed by drug-resistant parasites and rapid progression of malarial illness. This review article focuses on the progress achieved during the last years in the production of artemisinin from Artemisia annua. The structure, biosynthesis and analysis of artemisinin and its mode of action are described. The review also focuses on clinical studies, toxicity studies, pharmacokinetics and activity of artemisinin related compounds. The production strategies including organic synthesis, extraction from plants, in vitro cultures and alternative strategies for enhancing the yields are also discussed. PMID- 10665981 TI - Inhibition of inducible nitric oxide synthase in persistent pain. AB - The present study was undertaken to investigate the role of inducible nitric oxide synthase in a rat model of persistent pain. The effects of L-N6 (1 iminoethyl) lysine (L-NIL), a relatively potent and relatively selective inhibitor of inducible nitric oxide synthase, were investigated in carrageenan induced hyperalgesia L-NIL (0.1 microMole) injected intraplantar or intrathecal markedly enhanced carrageenan induced hyperalgesia. These effects were reversed during the third hour by co-administration of L-arginine (900 mg/kg i.p.) but not D-arginine. Methylene blue (MB), a soluble guanylate cyclase inhibitor, administered intrathecally (0.1 microg) had no effect on L-NIL potentiation of carrageenan hyperalgesia but abolished antinociception induced by L-arginine. Obtained results suggest that nitric oxide derived from inducible nitric oxide synthase play an inhibitory role in carrageenan produced hyperalgesia in rat. PMID- 10665982 TI - Enhanced pial arteriolar sensitivity to bioactive agents following exposure to endothelin-1. AB - Effects of prior exposure of pial arterioles to endothelin-1 (ET-1) (10(-9) M) on the constriction induced by the by-products of hemolyzed blood (5-HT, LTC4, LPA, and thromboxane analog U-46619) were examined. Piglets (age: 1-3 d) anesthetized with a mixture of ketamine hydrochloride and acepromazine were implanted with cranial windows, and anesthesia was maintained with alpha-chloralose. Topical applications of the by-products of hemolyzed blood mildly constricted pial arterioles. Following prior exposure of the microvessels to ET-1, application of the by-products of hemolyzed blood produced significantly potentiated and long lasting constrictions compared to the controls. In another experiment, pretreatment of pial arterioles with U-46619 (10(-8) M) also potentiated the constriction induced by ET-1. The constriction produced was fast and longer lasting. Thus, these data show that by-products of hemolyzed blood, though not potent vasoconstrictors per se, potently constricted pial arterioles in the presence of ET-1. The same agents in the CSF can also potentiate constriction induced by ET-1. Hence, by-products of hemolyzed blood may play a significant role in the initiation and maintenance of cerebral arterial narrowing observed following intracranial bleeding. PMID- 10665983 TI - Prolonged exposure to cigarette smoke blocks the neurotoxicity induced by kainic acid in rats. AB - We examined the effects of cigarette smoke (CS) on three parameters associated with kainic acid (KA)-induced neurotoxicity: seizure activity, cell loss in the hippocampus, and increased Fos-related antigen (FRA) expression. Animals were exposed to the main stream of CS from 15 Kentucky 2R1F research cigarettes containing 28.6 mg tar and 1.74 mg nicotine per cigarette, for 10 min a day, 6 days per week, for 4 weeks, using an automatic smoking machine. KA administration (10 mg/kg, i.p.) produced robust behavioral convulsions lasting 4-5 h. Pre exposure to CS significantly reduced the seizures, mortality, and severe loss of cells in regions CA1 and CA3 of the hippocampus after KA administration. Consistently, pre-exposure to CS significantly attenuated the KA-induced increased FRA immunoreactivity in the hippocampus. In contrast, pretreatment with central nicotinic antagonist, mecamylamine (2 or 10 mg/kg, i.p.) blocked the neuroprotective effects mediated by CS in a dose-dependent manner. These results indicate that CS exposure provides neuroprotection against the KA insult via nicotinic receptor activation. PMID- 10665984 TI - Mechanical characterization of regenerated osseous tissue during callotasis and its related biological phenomenon. AB - The mechanical characteristics of the regenerated osseous tissue and osteoblastic activities during callotasis were studied using Chinese mountain goat as the animal model. Open osteotomy of the left tibiae was done in 24 goats. Distraction started 6 days after the operation with the rate of 1 mm per day for 4 weeks. The bone regeneration was monitored with serial X-ray films. The tension generated during distraction was measured with the strain gauges mounted on the distraction apparatus. The osteoblastic activities were monitored by measuring plasma bone specific alkaline phosphatase activity. The results showed that the average lengthening was 22.9 +/- 2.8 mm in each animal. The newly formed osseous tissue becomes stiffer during the course of distraction lengthening. The continuous evolution of the tensile behaviour of the newly formed osseous tissue correlates with the plasma bone specific alkaline phosphatase activities. The radiological appearance of a physis like structure took place at 12 mm lengthening. Its appearance corresponds to the changes in the tensile behaviour as well as the biological activities of the osteoblasts and may serve as a useful radiological marker in monitoring the process of callotasis in clinical practice. PMID- 10665985 TI - Diabetes alters neuronal nitric oxide release from rat mesenteric arteries. Role of protein kinase C. AB - The objective of the present study was to assess the influence of diabetes in the neuronal nitric oxide (NO) release elicited by electrical field stimulation (EFS, 200 mA, 0.3 ms, 1-16 Hz, for 30 s, at 1 min interval) in endothelium-denuded mesenteric artery segments from control and streptozotocin-induced diabetic rats, assessing the influence of protein kinase C (PKC) in this release. N(G)-nitro-L arginine-methyl ester (L-NAME, 10 microM, a NO synthase inhibitor) enhanced EFS elicited contractions in control, and specially in diabetic rats, whereas they were unaltered by AMT (5 nM, an inducible NO synthase inhibitor) and capsaicin (0.5 microM, a sensory neurone toxin). Calphostin C (0.1 microM, a PKC inhibitor) increased the contraction elicited by EFS in both types of arteries. This increase was further enhanced by calphostin C + L-NAME in diabetic rats. Phorbol 12,13-dibutyrate (PDBu, 1 microM) reduced and unaltered EFS-induced contractions in control and diabetic rats, respectively. The further addition of L-NAME reversed the reduction obtained in control rats, and enhanced the response observed in diabetic rats. These results suggest that the EFS-induced NO release from perivascular nitrergic nerves, that negatively modulates the contraction, which is synthesized by neuronal constitutive NO synthase. The NO synthesis is positively stimulated by PKC. This NO release is increased in diabetes, likely due to an increase in the activity of this enzyme. The sensory nerves of these arteries do not seem to be involved in the contractile response. PMID- 10665986 TI - Effects of euxanthone on neuronal differentiation. AB - The growth inhibitory and differentiation inducing effects of euxanthone (1,7 dihydroxyxanthone) from the medicinal plant Polygala caudata on the neuroblastoma (Neural 2A, subclone BU-1) were investigated. At the concentration range of 0-100 microM, euxanthone inhibits the growth of BU-1 cells in a dose dependent manner. The 50% growth inhibitory concentration (IC50) was 41 microM. Significant induction of morphological differentiation and neurite growth was observed at the concentration of 100 microM. Frequency of proliferative neuroblastoma cells was determined after induction of differentiation. The frequency of proliferating BU 1 cells was markedly reduced from 1/1.1 to <1/99. Confocal microscopy also confirmed that the morphological differentiation of BU-1 was associated with the expression of neurite specific marker MAP-2 protein in neurites. These data suggest that euxanthone may be one of the neuropharmacological active compounds in the medicinal plant Polygala caudata. PMID- 10665987 TI - Kappa-opioid receptor stimulation increases the expression of Na+-H+ exchange gene in the heart. AB - Kappa-opioid receptor (OR) stimulation increases intracellular pH (pHi) via activating the Na+-H+ exchange (NHE). In the present study, we determined the expression of the gene of NHE1, the predominant NHE isoform in the heart, and intracellular pH (pHi) upon kappa-OR stimulation in the rat heart. We found that 1 microM U50,488H (trans-3,4-dichloro-N-methyl-N-(2-(1 pyrrolidinyl)cyclohexyl)benzeneacetamide), a selective kappa-OR agonist, increased the expression of the NHE1 gene. We also found that U50,488H dose dependently increased pHi in the heart. The effects were abolished by 1 microM nor-binaltorphimine (nor-BNI), a selective kappa-OR antagonist, indicating that the events were kappa-OR mediated. The effects on both NHE1 gene expression and pHi were also abolished by 5 microM chelerythrine and 5 microM BSM (bisyndolylmaleimide), protein kinase C (PKC) inhibitors, indicating that PKC mediated the actions. In addition, the effect of U50,488H on pHi was blocked by 10 microM EIPA (ethylisopropyl amiloride), a NHE1 inhibitor, indicating that NHE1 also mediated the action of U50,488H. The present study provides evidence for the first time that kappa-OR stimulation increased the NHE1 gene expression in the heart via a PKC dependent pathway. Kappa-OR stimulation also increases pHi via PKC and NHE in the heart. PMID- 10665988 TI - Enterohepatic circulation of chloramphenicol and its glucuronide in the rat by microdialysis using a hepato-duodenal shunt. AB - A system consisting of a hepato-duodenal shunt in which the bile of a drug treated donor rat was diverted to the duodenum of an untreated recipient rat via a bile cannula was used to assess the role of hepatic metabolism and enterohepatic circulation in the pharmacokinetics of chloramphenicol. Blood concentrations of unbound chloramphenicol and its glucuronide were measured by on line microdialysis coupled to a microbore liquid chromatographic system. Results indicated that chloramphenicol and its glucuronide were detected in the blood of both donor and recipient rats following an intravenous 100 mg/kg dose of chloramphenicol succinate to the donor rat. Our finding suggests that although enterohepatic circulation contributed only to a minor extent (approximately 1.8%) was involved in the disposition of unbound chloramphenicol in the rat on-line microdialysis techniques were applicable for such studies. PMID- 10665990 TI - Molluscicidal and piscicidal activities of Venezuelan Chrysobalanaceae plants. AB - Extracts of increasing polarity of 6 Venezuelan plants belonging to the Chrysobalanaceae family were tested for possible letal toxicity against Biomphalaria glabrata Say, a snail intermediate hosts of Schistosoma mansoni Sambon. The piscicidal toxicity of the active extracts was also evaluated with the aim of findings compounds without toxicity in nontarget organisms, principally fishes and humans. PMID- 10665989 TI - Increased brain P-glycoprotein in morphine tolerant rats. AB - The objective of this study was to determine whether chronic morphine exposure increased P-glycoprotein in rat brain. Male Sprague-Dawley rats were treated with morphine, saline, or dexamethasone for 5 days. On day 6, antinociceptive effect was measured to evaluate the extent of functional tolerance to morphine. Brain P glycoprotein was detected by Western blot analysis of whole brain homogenate. Morphine- and dexamethasone-treated rats exhibited decreased antinociceptive response when compared to saline-treated controls. Brain P-glycoprotein was approximately 2-fold higher in morphine-treated rats compared to saline controls based on Western blot analysis. Chronic morphine exposure appears to increase P glycoprotein in rat brain. P-glycoprotein induction may enhance morphine efflux from the brain, thus reducing morphine's pharmacologic activity. Induction of P glycoprotein may be one mechanism involved in the development of morphine tolerance. PMID- 10665991 TI - Differential potentiative effects of GABA receptor agonists in the production of antinociception induced by morphine and beta-endorphin administered intrathecally in the mouse. AB - The effect of muscimol or baclofen injected intrathecally (i.t.) on the inhibition of the tail-flick response induced by morphine and beta-endorphin administered i.t. was studied in ICR mice. The i.t. injection of muscimol (100 ng) or baclofen (10 ng) alone did not affect the basal inhibition of the tail flick response. Morphine (0.2 microg) and beta-endorphin (0.1 microg) caused only slight inhibition of the tail-flick response. Baclofen, but not muscimol, injected i.t. enhanced the inhibition of the tail-flick response induced by i.t. administered morphine. Both muscimol and baclofen injected i.t. significantly enhanced i.t. injected beta-endorphin-induced inhibition of the tail-flick response. Our results suggest that the GABA(B), but not GABA(A), receptors located in the spinal cord appear to be involved in enhancing the inhibition of the tail-flick response induced by morphine administered spinally. In addition, both GABA(A) and GABA(B) receptors are involved in enhancing the inhibition of the tail-flick response induced by beta-endorphin administered i.t. PMID- 10665992 TI - Toxicity during early development of the mouse nervous system of a scorpion neurotoxin active on sodium channels. AB - The lethal effects of scorpion envenomation is due to neurotoxins active on voltage-sensitive sodium channels. Dysfunctions of the peripheral and central nervous systems with neurological manifestations are commonly observed after scorpion stings, specially in young children. Since the neurotoxicity of venom fraction is greatly higher by intracerebroventricular than by subcutaneous injections, a direct effect of venom on CNS cannot be excluded specially in infants where the blood-brain barrier is not fully functional. We investigated the activity of a neurotoxin from the scorpion Androctonus australis hector (AahII) in newborn mice at 3, 7 and 14 days after birth and in adults. Young mice (P3, P7) were more sensitive to AahII injected subcutaneously than were adults, but were less sensitive to intracerebroventricular injection. The affinity of AahII for its receptor site on brain synaptosomes from P3 and P7 mice was slightly higher and the density of the binding sites was half that of adult mice. After subcutaneous injection of [125I]-AahII it was also observed that a small amount of radioactivity was found in brains of neonate mice but not in that of adults. This amount is however extremely lower than the value of the LD50 determined by intracerebroventricular injection. Results are consistent with a peripheral action of AahII and show that its toxic activity changes during the mouse nervous system development. PMID- 10665993 TI - Response of high mobility group proteins of human kidney T1 and murine L 929 cell lines to heat shock. AB - High mobility group (HMG) proteins in human kidney T1 and murine L 929 cells have been investigated after exposure to heat shock at 41 degrees C and their influence on the organizational change of chromatin under heat shock condition has been examined. Results reveal that the two cell lines show differential response of the HMG proteins 1 & 2 and 14 & 17 to heat shock. Neither T1 nor L 929 cells show significant differences in response to heat shock with respect to the binding affinities of HMG proteins 1 & 2 or 14 & 17 to DNA, as revealed by DNase I sensitivity and chromatin reconstitution assays. Furthermore, the HMG proteins of both the non-heat shocked and the heat shocked T1 and L 929 cells can recover their chromatin activity following reconstitution. These findings suggest that although the HMG proteins might undergo some change in response to heat shock, their inherent potential of reassociation with DNA is still retained. PMID- 10665994 TI - In vivo exposure to carbon disulfide increases the contraction frequency of pregnant rat uteri through an indirect pathway. AB - Exposure to CS2, an organic solvent, is associated with an increased rate of abnormal labor or dysmenorrhea. Contraction of quiescent uteri during pregnancy can cause preterm labor. We wish to know the effects of in vivo and in vitro exposures to CS2 on uterine contractions of mid-gestation rats. After 10-d exposure to 300 or 600 mg/kg CS2, uteri of pregnant rats were measured for contractile responses to various stimuli, such as KCl, oxytocin, carbachol or A23187, a calcium ionophore, using standard muscle bath apparatus. CS2 treatment significantly increased the contractile response to KCl, carbachol, and A23187. The increase to A23187 was the greatest. In contrast, in vitro exposure to CS2 immediately suppressed carbachol-induced contraction but did not affect spontaneous and KCl-induced contractions. Results showed the pregnant uterus of the rat is susceptible to CS2. The influence of in vivo exposure to CS2 on uterine contraction was opposite to that in vitro. The increased response of CS2 treated uteri to A23187 suggests that in vivo exposure to CS2 may sensitize contraction machinery to calcium through indirect pathways. PMID- 10665995 TI - Circadian periodicity of urinary volume, creatinine and 5-hydroxyindole acetic acid excretion in healthy Indians. AB - The circadian periodicity of urinary output, creatinine (Cr) and 5-hydroxyindole acetic acid (5-HIAA) excretion was studied under near-tropical conditions in 130 healthy volunteers (65 men and 65 women, 16-75 years of age) with a diurnal activity from about 06:30 to about 22:00 and nocturnal rest. These volunteers were divided into 4 groups, 16-30, 31-45, 46-60 and 61-75 years of age, comprising 20, 20, 15 and 10 participants of each gender, respectively. A marked circadian rhythm was recorded for urine volume, Cr and 5-HIAA excretion in healthy Indians of different ages. The acrophase tended to be delayed in the older age group. The relative amplitude decreased with advancing age, notably in women. Overall, men produced a larger urine volume as compared to women. Excretions of Cr and 5-HIAA in healthy Indian volunteers of different ages may be influenced by diet, societal relations, climate and/or geographic location. The contribution of such factors in metabolism and degradation warrants further study. PMID- 10665996 TI - Shear stress enhances prostacyclin release from endocardial endothelial cells. AB - The effect of shear stress on the release of prostacyclin (PGI2) from cultured endocardial endothelial cells (EECs) was investigated. EECs were harvested from the right ventricle (RV) and the left ventricle (LV) of porcine heart. Confluent EECs were incubated under various degrees of shear stress (0.2, 1, 4 and 6 dyne/cm2) and PGI2 release from each cell was measured. PGI2 release from LV-EECs and RV-EECs was enhanced by the elevation of shear stress in a shear-dependent manner with a rapid increase at the onset of flow; however, there was no significant difference in PGI2 production between RV-EECs and LV-EECs. production of PGI2 was significantly inhibited from cells exposed to 8-(dimetilamino) octyl 3,4,5-trymethoxybenzoate hydrochloride (10 and 100 microM: an inhibitor of intracellular calcium mobilization) or cyclopiazonic acid (10 microM: an endoplasmic reticulum Ca2+-ATPase inhibitor). These results indicate that shear stress enhances PGI2 release from cultured EECs and that mechanotransduction of shear stress depends on calcium mobilization in EECs. PMID- 10665997 TI - Lack of interaction in recombinant human FSH receptor and both TSAb and TSBAb. AB - Since cross-reactivity of TSH with the human FSH receptor has been reported, in this study we tested the effect of thyroid-stimulating antibody (TSAb) and thyroid stimulation-blocking antibody (TSBAb) on Chinese hamster ovary cells expressing human FSH receptor (CHO-hFSH-R cells). We examined the TSBAb activity of sera from hypothyroid patients who had a positive TBII to determine whether these sera also block the effect of FSH on CHO-hFSH-R cells. Although human FSH I 3 (0.25-16 ng/ml) stimulated the production of intracellular cAMP in CHO-hFSH-R cells with dose-responsive manner, neither TSAb nor TSBAb had such an effect on the cells. PMID- 10665998 TI - Uncoupling effect of anacardic acids from cashew nut shell oil on oxidative phosphorylation of rat liver mitochondria. AB - Anacardic acids are one of natural products found in not only the cashew nut shell oil but also the nut and fruit juice. The present study was conducted to investigate the uncoupling effect of anacardic acids on oxidative phosphorylation of rat liver mitochondria using succinate (plus rotenone) as a substrate. Four anacardic acids with C15:0, C15:1, C15:2 or C15:3 as an alkyl side chain exhibited uncoupling effects similar to the classical uncoupler, 2,4 dinitrophenol on ADP/O ratio, state 4 and respiratory control ratio (RCR). Anacardic acid with C15:1 side chain was most effective for uncoupling of these compounds. Salicylic acid, which has no alkyl side chain, exhibited a very weak uncoupling effect on oxidative phosphorylation. When the carboxyl group in anacardic acids was lost converting them to the corresponding cardanols, uncoupling activity dramatically decreased regardless of the number of double bonds in the long alkyl chain. These results suggest that the C15 alkyl side chain as well as the carboxyl group may play an important role in assisting the uncoupling activity of anacardic acids in liver mitochondria of animals. This study provides the first evidence of an uncoupling effect of anacardic acids on liver mitochondria PMID- 10665999 TI - Modulation of collagen synthesis by tumor necrosis factor alpha in cultured vascular smooth muscle cells. AB - Collagen synthesis in vascular smooth muscle cells (SMCs) after exposure to tumor necrosis factor alpha (TNF-alpha) was investigated using a culture system. The synthesis of collagenase-digestible proteins (CDP) and noncollagenous proteins (NCP) was evaluated by the [3H]proline incorporation. It was shown that TNF-alpha markedly suppresses the incorporation of [3H]proline into both CDP and NCP in confluent cultures of SMCs but not in sparse cultures of the cells. Such a marked suppression by TNF-alpha was not observed in confluent bovine aortic endothelial cells and human fibroblastic IMR-90 cells. In confluent SMCs, the synthesis of CDP was more strongly inhibited by TNF-alpha than that of NCP. When the CDP synthesis was stimulated by transforming growth factor beta, TNF-alpha suppressed the stimulation in both confluent and sparse SMCs. Human SMCs synthesized types I, III, IV and V collagens; TNF-alpha markedly decreased the relative proportion of types IV and V. It was therefore suggested that TNF-alpha modulates the collagen synthesis by SMCs depending on their cell density and modifies the formation of atherosclerotic lesions. PMID- 10666000 TI - Tumor necrosis factor-alpha-mediated signal transduction in human neutrophils: involvement of sphingomyelin metabolites in the priming effect of TNF-alpha on the fMLP-stimulated superoxide production. AB - We investigated the mechanism underlying the priming effect of TNF-alpha on fMLP stimulated superoxide production in human neutrophils. TNF-alpha enhanced fMLP stimulated superoxide production in a concentration-dependent manner. TNF-alpha also induced sphingomyelin (SM) hydrolysis and increased the formation of its metabolite, sphingosine-1-phosphate (SP-1-P). The treatment of neutrophils with sphingomyelinase also resulted in a similar priming effect. C2 ceramide produced a concentration-dependent inhibition of fMLP-stimulated superoxide production within the concentration range of 1-30 microM. Sphingosine had a dual effect on fMLP-stimulated superoxide generation, exhibiting a priming effect at lower concentrations (0.2-1 microM), but an inhibitory effect at higher concentrations (1-30 microM). SP-1-P (1-30 microM), showed a concentration-dependent enhancement of fMLP stimulated superoxide production. Furthermore, after treating neutrophils with DL-threo-dihydro-sphingosine, a competitive inhibitor of sphingosine kinase, TNF-alpha produced a similar dual effect as observed with sphingosine. These results strongly suggest that SM hydrolysis plays a key role in the intracellular signal transduction mediating the TNF-alpha-mediated priming effect. PMID- 10666001 TI - Supraspinal and spinal effects of [Phe1psi(CH2-NH)Gly2]-nociceptin(1-13)-NH2 on nociception in the rat. AB - A new derivative of the neuropeptide nociceptin (NC) has recently been developed. This molecule, the pseudopeptide [Phe1psi(CH2-NH)Gly2]-nociceptin(1-13)-NH2 was found to antagonize NC inhibitory effects in peripheral smooth muscle preparations in vitro. However, contrasting results have appeared as regards its pharmacodynamic profile in the CNS. Here, we investigated the pseudopeptide effects, in vivo, on nociceptive responses in the rat. [Phe1psi(CH2-NH)Gly2] nociceptin(1-13)-NH2 was administered intracerebroventricularly (i.c.v.) or intrathecally (i.t.) (alone or in combination with NC), and tail-flick latencies (TFL) to radiant heat were assessed. I.c.v. [Phe1psi(CH2-NH)Gly2]-nociceptin(1 13)-NH2 (1-10 nmol/rat) caused a short-lasting decrease (5 min) of TFL and did not antagonize the threshold lowering effect of i.c.v. NC (1 nmol/rat). At the spinal level, the i.t. administration (0.2-10 nmol/rat) of [Phe1psi(CH2-NH)Gly2] nociceptin(1-13)-NH2 produced a dose-dependent and long-lasting antinociceptive effect that was not modified by the administration of a high dose (30 nmol/rat i.t.) of the opioid antagonist naloxone. The i.t. co-administration of the pseudopeptide (10 nmol/rat) did not block the antinociceptive effect of i.t. NC (10 nmol/rat). These data indicate that the pseudopeptide behaves as an NC agonist at supraspinal and spinal levels in the rat tail-flick test of nociception. These different profiles in the periphery and the CNS could suggest differences between central and peripheral NC receptor/s and provide a basis for further development of antagonist molecules suitable for their characterization. PMID- 10666002 TI - Protective effect of aminoguanidine, a nitric oxide synthase inhibitor, against carbon tetrachloride induced hepatotoxicity in mice. AB - The present study was undertaken to evaluate the effect of aminoguanidine (AG) on carbon tetrachloride (CCl4)-induced hepatotoxicity. Treatment of mice with CCl4 (20 microl/kg, i.p.) resulted in damage to centrilobular regions of the liver, increase in serum aminotransferase and rise in lipid peroxides level 24 hours after CCl4 administration. Pretreatment of mice with AG (50 mg/kg, i.p.) 30 minutes before CCl4 was found to protect mice from the CCl4-induced hepatic toxicity. This protection was evident from the significant reduction in serum aminotransferase, inhibition of lipid peroxidation and prevention of CCl4-induced hepatic necrosis revealed by histopathology. Aminoguanidine, a relatively specific inhibitor of inducible nitric oxide synthase, did not inhibit the in vitro lipid peroxidation. Taken together, these data suggest a potential role of nitric oxide as an important mediator of CCl4-induced hepatotoxicity. PMID- 10666003 TI - The inhibition of GLUT1 glucose transport and cytochalasin B binding activity by tricyclic antidepressants. AB - Under normal metabolic conditions glucose is an important energy source for the mammalian brain. Positron Emission Tomography studies of the central nervous system have demonstrated that tricyclic antidepressant medications alter cerebral metabolic function. The mode by which these drugs perturb metabolism is unknown. In the present study the interactions of tricyclic antidepressants with the GLUT1 glucose transport protein is examined. Amitriptyline, nortriptyline, desipramine, and imipramine all inhibit the influx of 3-O-methyl glucose into resealed erythrocytes. This inhibition is observed with drug concentrations in the millimolar range. All four antidepressants also noncompetitively displace cytochalasin B binding to GLUT1. The K(I) for this displacement ranges from 0.56 to 1.43 millimolar. This value is in a range greater than that associated with clinical doses and this effect may not be directly applicable to side effects observed with normal use. The observed interaction of these drugs with GLUT1 may reflect an affinity for other glucose-transport or glucose-binding proteins, and may possibly contribute to tricyclic antidepressant toxicity. PMID- 10666004 TI - Roles of endogenous prostaglandins and nitric oxide in inhibitions of gastric emptying and accelerations of gastrointestinal transit by escins Ia, Ib, IIa, and IIb in mice. AB - We reported previously that escins Ia, Ib, IIa, and IIb, isolated from horse chestnuts, inhibited the 30-min gastric emptying (GE) in mice. In this study, the effects of escins Ia-IIb on gastrointestinal transit (GIT), and the roles of endogenous prostaglandins (PGs) and nitric oxide (NO) in the effects of escins Ia -IIb on GE and GIT were investigated in fasted mice. Escins Ia-IIb (12.5-50 mg/kg, p.o.) dose-dependently accelerated GIT. Both GE inhibitions and GIT accelerations by escins Ia-IIb (25 mg/kg) were markedly attenuated by pretreatment with indomethacin (10 mg/kg, s.c., an inhibitor of PGs synthesis). Pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME, 10 mg/kg, i.p., an inhibitor of constitutive and inducible NO synthase) attenuated the effects of escins Ia-IIb on GIT, but not on GE. The effect of L-NAME was reversed by L arginine (600 mg/kg, i.p., a substrate of NO synthase), but not by D-arginine (900 mg/kg, i.p., the enantiomer of L-arginine). The GIT accelerations of escins Ia-IIb were not attenuated by pretreatment with D-NAME (10 mg/kg, i.p., the enantiomer of L-NAME) or dexamethasone (5 mg/kg, i.p., an inhibitor of inducible form of NO synthase). The results suggest that endogenous PGs play an important role in both GE inhibitions and GIT accelerations, and constitutive NO is involved in the GIT accelerations, by escins Ia--IIb in mice. PMID- 10666005 TI - GIP biology and fat metabolism. AB - The gastrointestinal hormone, gastric inhibitory polypeptide (GIP), is synthesized and released from the duodenum and proximal jejunum postprandially. Its release depends upon several factors including meal content and pre-existing health status (ie. obesity, diabetes, age, etc.). It was initially discovered and named for its gastric acid inhibitory properties. However, its more physiologically relevant role appears to be as an insulinotropic agent with a stimulatory effect on insulin release and synthesis. Accordingly, it was later renamed glucose-dependent insulinotropic polypeptide because its action on insulin release depends upon an increase in circulating levels of glucose. GIP is considered to be one of the principle incretin factors of the enteroinsular axis. The GIP receptor is a G-protein-coupled receptor belonging to the family of secretin/VIP receptors. GIP receptor mRNA is widely distributed in peripheral organs, including the pancreas, gut, adipose tissue, heart, adrenal cortex, and brain, suggesting it may have other functions in addition to the ones mentioned above. An overactive enteroinsular axis has been suggested to play a role in the pathogenesis of diabetes and obesity. In addition to stimulating insulin release, GIP has been shown to amplify the effect of insulin on target tissues. In adipose tissue, GIP has been reported to (1) stimulate fatty acid synthesis, (2) enhance insulin-stimulated incorporation of fatty acids into triglycerides, (3) increase insulin receptor affinity, and (4) increase sensitivity of insulin-stimulated glucose transport. In addition, although controversial, lipolytic properties of GIP have been proposed. The mechanism of action of GIP-induced effects on adipocytes is unknown, and it is unclear whether these effects of GIP on adipocytes are direct or indirect. However, there is now evidence that GIP receptors are expressed on adipocytes and that these receptors respond to GIP stimulation. Given the location of its release and the timing of its release, GIP is an ideal anabolic agent and expanding our understanding of its physiology will be needed to determine its exact role in the etiology of diabetes mellitus and obesity. PMID- 10666006 TI - Electroencephalographic effects of thiopentone and its enantiomers in the rat. AB - Electrophysiological studies with some chiral barbiturates have shown that one enantiomer can be excitant while the other is depressant. Thiopentone, a chiral barbiturate, has both differences in potency between enantiomers and biphasic effects on the electroencephalogram (EEG). This study investigated whether a differential EEG activity between the enantiomers of thiopentone could account for the biphasic effects. Rats were administered rac-, R- or S-thiopentone to determine the nature and time course of quantitative EEG effects. Two studies using computer-controlled i.v. infusions of the three drugs were performed in groups of animals previously prepared with EEG electrodes and/or arterial blood sampling cannulae. Study 1 used several stepwise increments in plasma drug concentration over 35 min, followed by washout. Study 2 used a 4 min period of constant plasma drug concentration, followed by washout. In both studies, both enantiomers and racemate caused an initial EEG activation followed by deactivation. Quantitative enantioselectivity was found for depression. The extent of depression was significantly less for R-thiopentone (P=0.008) and racthiopentone (P=0.038) than for S-thiopentone; recovery from depression appeared to be faster for R-thiopentone than either rac- or S-thiopentone. Fatality was only found with S-thiopentone (3/7 animals in Study 2). R thiopentone plasma concentrations were approximately 8% less than those of S thiopentone in rats treated with racthiopentone. Although small differences in clearance between enantiomers were found that may influence recovery, they were not large enough to account for the reported differences in potency between the two enantiomers. PMID- 10666007 TI - Soy based diet attenuates the development of hypertension when compared to casein based diet in spontaneously hypertensive rat. AB - The purpose of this study was to compare the effects of soy and casein based diets on blood pressure and cardiovascular functions in male and female spontaneously hypertensive rats (SHR). The systolic blood pressure was measured at the beginning and at the end of study. After a five week supplementation period with three different diets, the rats were decapitated and arterial responses and the weight-to-body weight-ratios of the organs were studied. The development of hypertension was attenuated in both female and male rats on soy protein diet when compared to the casein diet. Soy based diet lowered serum total cholesterol level when compared to the control diet. Both casein and soy protein supplementation in diet induced a significant renal hypertrophy in both female and male SHR rats when compared to SHR rats on the control diet. Soy protein supplementation reduced significantly serum estradiol-17beta concentration when compared to the control diet. There were no differences in the serum testosterone concentrations between the diet groups. When compared to the casein based diet the soy based diet attenuated the development of hypertension and decreased serum total cholesterol level in SHRs. These effects were independent of gender. The mechanisms and clinical importance of these findings remain to be clarified. PMID- 10666009 TI - Expression of adhesion receptors on rat limb bud cells and results of treatment with a thalidomide derivative. AB - The expression of several adhesion surface receptors was studied on cells of early limb bud development of 58 Wistar rats treated orally with two daily doses of the thalidomide derivative EM12 (2 x 50 mg/kg body weight) from day 7 to 10 of pregnancy. EM12 is a more potent teratogen than thalidomide. Limb bud cells of 56 untreated animals served as controls. The studies revealed that the integrins CD11a, CD11b, CD18, CD49d, and CD61, as well as the additional adhesion receptors CD54, CD62L, and the transferrin receptor CD71 were expressed on day 11 of gestation to various degrees on these embryonic cells. In contrast to results of previous studies with a non-human primate (Callithrix jacchus) there was no down regulation of any of these receptors on the surface of limb bud cells of the rat embryos after treatment with EM12. This result is in accordance with the lack of teratogenicity in this rodent species. PMID- 10666008 TI - Fatty liver and hyperlipidemia in IDDM (insulin-dependent diabetes mellitus) of streptozotocin-treated shrews. AB - Severe IDDM (insulin-dependent diabetes mellitus) was produced in the musk shrew (Suncus murimus, Insectivora) by a high dose (a single intraperitoneal injection of 100 mg/kg Body Weight) of streptozotocin (STZ) injection. All shrews that were administered a high dose of STZ exhibited hyperglycemia (449 +/- 16 mg/dl vs 73 +/- 4 mg/dl in controls) and hypoinsulinemia(0.25 +/- 0.07 ng/ml vs 10.96 +/- 1.97 ng/ml in controls) with ketosuria 10 days after injection. Their livers were enlarged and exhibited ayellowish-brown color with marked triglyceride (TG) accumulation (63.25 +/- 7.10 mg/g Liver vs 2.11 +/- 0.19 mg/g Liver in controls). It is probable that the increased influx of fatty acids into the liver induced by hypoinsulinemia and the low capacity of excretion of lipoprotein secretion from liver in the musk shrew resulting from a deficiency of apolipoprotein B synthesis play important roles in fatty liver formation. Hyperlipidemia was another feature in shrews with severe IDDM. The blood TG level was especially high in these shrews (899 +/- 178 mg/dl vs 23 +/- 5 mg/dl in controls). These results indicate that the IDDM shrew, induced by high doses of STZ, is a unique model characterized by fatty liver and hyperlipidemia and may be useful for studying lipid metabolism of IDDM. PMID- 10666010 TI - Unexpected suppression of free phenytoin concentration by salicylate in uremic sera due to the presence of inhibitors: MALDI mass spectrometric determination of molecular weight range of inhibitors. AB - Salicylate displaces phenytoin from protein binding leading to an increase in free phenytoin concentration. We observed unexpected decreases in free phenytoin concentration in the presence of salicylate. Serum pools containing no phenytoin or salicylate were supplemented with the same concentrations of phenytoin. Then to the aliquots of the individual pool, no salicylate (control), 150, 300 and 500 microg/ml of salicylate (therapeutic range: 15-300 microg/ml) were added. Specimens were incubated at 37 degrees C for 2 h and after re-equilibration at room temperature for 20 min, total and free phenytoin (in the protein free ultrafiltrates) concentrations were measured using fluorescence polarization immunoassay on the TDx/FLX analyzer. We observed an increase in free phenytoin concentration from 1.91 microg/ml (in the absence of salicylate) to 2.39 microg/ml in the presence of 500 microg/ml salicylate (total phenytoin: 13.3 microg/ml) in the normal pool. In sharp contrast, the free phenytoin concentrations decreased from an initial concentration of 3.82 microg/ml to 2.52 microg/ml in the presence of 500 microg/ml of salicylate (total phenytoin: 13.2 microg/ml) in the uremic pool. We also treated the uremic pool with activated charcoal. In the original uremic pool, the initial free phenytoin concentration was 3.05 microg/ml and the free concentrations then decreased to 2.28 microg/ml in the presence of 300 microg/ml of salicylate. In contrast, in the charcoal treated pool, the initial free phenytoin concentration increased from 1.61 microg/ml to 3.23 microg/ml in the presence of 300 microg/ml of salicylate. More interestingly when uremic toxins were extracted back from charcoal with methanol and the dry residue was added to an aliquot of normal serum, the normal serum behaved like a uremic serum and free phenytoin concentration was significantly decreased in the presence of salicylate. When an aliquot of methanol extract was studied by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry (scan up to 10,000), we observed no peak at molecular weight over 551, indicating that these inhibitors are small molecules. We also identified hippuric acid as one of the inhibitors. PMID- 10666011 TI - Melatonin increases muscle and liver glycogen content in nonexercised and exercised rats. AB - The effects of melatonin on several parameters of carbohydrate and lipid metabolism were investigated in exercised and nonexercised rats. Animals were run to exhaustion on a rodent treadmill at 24 m/min and a 12% slope. Exercise resulted in a significant hypoglycemia and increased plasma levels of lactate and beta-hydroxybutyrate, together with a significant reduction of glycogen in muscle and liver. Muscle and liver glycogen content was elevated and plasma free fatty acid decreased in nonexercised animals receiving melatonin (0.5 or 2.0 mg/kg i.p). Melatonin at 2.0 mg/kg reduced plasma lactate and increased lactate concentration in liver. When compared to untreated exercised animals glycemia and muscle and liver glycogen content were significantly higher in melatonin-treated exercised animals, while plasma and liver lactate and plasma beta-hydroxybutyrate were significantly reduced. Our data indicate that melatonin preserves glycogen stores in exercised rats through changes in carbohydrate and lipid utilization. PMID- 10666012 TI - Mode of action of sesame lignans in protecting low-density lipoprotein against oxidative damage in vitro. AB - We investigated the antioxidant properties of sesaminol, a major component of sesame oil, on the oxidative modification of human low-density lipoprotein (LDL) in vitro. Sesaminol inhibited the Cu2+-induced lipid peroxidation in LDL in a concentration-dependent manner with an IC50 36.0 +/- 10.0 nM. Sesaminol was a more effective scavenger than either alpha-tocopherol or probucol in reducing the peroxyl radicals derived from 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) in aqueous solution. In addition, as determined by the secondary products of lipid peroxidation identified by using immunochemical methods, sesaminol completely inhibited the formation of 4-hydroxy-nonenal (4-HNE)- and malondialdehyde (MDA)-adducts in a concentration-dependent manner. Probucol and alpha-tocopherol at the same concentration exhibited a lesser inhibitory effect. Our findings suggest that sesaminol is a potentially effective antioxidant that can protect LDL against the oxidation. PMID- 10666013 TI - Negative NO3- difference in human coronary circulation with severe atherosclerotic stenosis. AB - To examine whether or not the levels of NOx (nitrite; NO2- and nitrate; NO3-) in coronary circulating blood reflect endothelial dysfunction due to coronary atherosclerosis, NOx levels in plasma obtained from ostium of left coronary artery and coronary sinus of patients who complained of chest pain were evaluated in relation to their coronary angiographic findings. Prior to the study, a HPLC Griess system for NOx measurement was critically evaluated. This system has a detection limit of 0.1 microM of NO2- and NO3- by 10 microl of loading and was able to distinguish a difference of 0.1-0.2 microM of these substances. Heparin (1 U/10 microl) did not affect the detective and discriminative abilities. NO3- difference, calculated from sino-arterial difference of NO3-, was almost zero ( 0.2 +/- 0.2 microM) in patients with either normal coronary arteries or mild organic coronary stenosis (< or = 20% narrowing), while a significant negative value (-5.9 +/- 1.7 microM) was obtained from patients with significant stenosis (> or = 70% narrowing) in the left coronary arteries. These results demonstrate reliable ability on the HPLC-Griess system in evaluating NO2- and NO3- in biological samples, and that the negative NO3- difference through coronary circulation may reflect endothelial dysfunction in the patients with coronary atherosclerosis with severe organic stenosis. PMID- 10666014 TI - Pulmonary protective effects of curcumin against paraquat toxicity. AB - An early feature of paraquat (PQ) toxicity is the influx of inflammatory cells, releasing proteolytic enzymes and oxygen free radicals, which can destroy the lung epithelium and result in pulmonary fibrosis. Therefore, the ability to suppress early lung injury seems to be an appropriate therapy of pulmonary damage before the development of irreversible fibrosis. Here I show curcumin confers remarkable protection against PQ lung injury. A single intraperitoneal injection of PQ (50 mg/kg) resulted in a significant rise in the levels of protein, angiotensin converting enzyme (ACE), alkaline phosphatase (AKP), N-acetyl-beta-D glucosaminidase (NAG) and thiobarbituric acid reactive substances (TBARS), and neutrophils in the bronchoalveolar lavage fluid (BALF), while a decrease in glutathione levels. In paraquat rats bronchoalveolar lavage (BAL) cell TBARS concentration was increased with a simultaneous decrease in glutathione content. In addition, the data also demonstrated that PQ caused a decrease in ACE and glutathione levels and an increase in levels of TBARS and myeloperoxidase (MPO) activity in the lung. Interestingly, curcumin prevented the general toxicity and mortality induced by PQ and blocked the rise in BALF protein, ACE, AKP, NAG TBARS and neutrophils. Similarly, curcumin prevented the rise in TBARS content in both BAL cell and lung tissue and MPO activity of the lung. In addition, PQ induced reduction in lung ACE and BAL cell and lung glutathione levels was abolished by curcumin treatment. These findings indicate that curcumin has important therapeutic implications in facilitating the early suppression of PQ lung injury. PMID- 10666015 TI - Y-27632 inhibits gastric motility in conscious rats. AB - Y-27632, a highly selective inhibitor of p160ROCK, desensitizes the smooth muscle to Ca2+ and inhibits smooth muscle contraction. While this drug has the potential to become a novel drug for hypertension, it might also affect other smooth muscle, including that of gastrointestinal tract. We studied the effects of Y 27632 on gastric contractions in conscious rats. Strain gauge force transducers were sutured onto the serosal side of the gastric antrum and contractions were recorded before and after the intravenous injection of Y-27632. Doses of 1.0 mg/kg to 10 mg/kg significantly decreased contraction amplitude and the motility index in a dose dependent manner. With 10 mg/kg, the mean amplitude was decreased by up to 69 +/- 14% and the motility index by up to 81 +/- 7%. The change occurred immediately after drug infusion and lasted for 3.5h. Contraction frequency showed only a slight decrease. No signs of bowel obstruction were observed. These results indicate that Rho-mediated Ca sensitization has a role in the physiologic contractions of gastric smooth muscle in rats. Y-27632 is useful to investigate the physiology of gastrointestinal motility. PMID- 10666017 TI - Unravelling the spindle PMID- 10666016 TI - Anti-phospholipase A2 and anti-inflammatory activity of Santolina chamaecyparissus. AB - The activity of the Santolina chamaecyparissus methanol extract was tested against the phospholipase A2 (PLA2)-induced mouse paw edema and in vitro inhibition of PLA2 activity. After fractionation, only the dichloromethane extract was active against the PLA2 in vitro test. In addition, it reduced the edema induced by arachidonic acid, and by 12-O-tetradecanoylphorbol-13-acetate in a multidose test. After chromatography on silicagel and gel filtration on Sephadex, and using an in vitro anti-PLA2 assay-guided process, we have isolated and identified from the dichloromethane extract the flavone nepetin and four sesquiterpenes. PMID- 10666018 TI - Calcium sparks in smooth muscle. AB - Local intracellular Ca(2+) transients, termed Ca(2+) sparks, are caused by the coordinated opening of a cluster of ryanodine-sensitive Ca(2+) release channels in the sarcoplasmic reticulum of smooth muscle cells. Ca(2+) sparks are activated by Ca(2+) entry through dihydropyridine-sensitive voltage-dependent Ca(2+) channels, although the precise mechanisms of communication of Ca(2+) entry to Ca(2+) spark activation are not clear in smooth muscle. Ca(2+) sparks act as a positive-feedback element to increase smooth muscle contractility, directly by contributing to the global cytoplasmic Ca(2+) concentration ([Ca(2+)]) and indirectly by increasing Ca(2+) entry through membrane potential depolarization, caused by activation of Ca(2+) spark-activated Cl(-) channels. Ca(2+) sparks also have a profound negative-feedback effect on contractility by decreasing Ca(2+) entry through membrane potential hyperpolarization, caused by activation of large conductance, Ca(2+)-sensitive K(+) channels. In this review, the roles of Ca(2+) sparks in positive- and negative-feedback regulation of smooth muscle function are explored. We also propose that frequency and amplitude modulation of Ca(2+) sparks by contractile and relaxant agents is an important mechanism to regulate smooth muscle function. PMID- 10666019 TI - Focus on "Sodium 4-phenylbutyrate downregulates Hsc70: implications for intracellular trafficking of DeltaF508-CFTR". PMID- 10666020 TI - Sodium 4-phenylbutyrate downregulates Hsc70: implications for intracellular trafficking of DeltaF508-CFTR. AB - The most common mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), DeltaF508, is a trafficking mutant that has prolonged associations with molecular chaperones and is rapidly degraded, at least in part by the ubiquitin-proteasome system. Sodium 4-phenylbutyrate (4PBA) improves DeltaF508-CFTR trafficking and function in vitro in cystic fibrosis epithelial cells and in vivo. To further understand the mechanism of action of 4PBA, we tested the hypothesis that 4PBA modulates the targeting of DeltaF508-CFTR for ubiquitination and degradation by reducing the expression of Hsc70 in cystic fibrosis epithelial cells. IB3-1 cells (genotype DeltaF508/W1282X) that were treated with 0.05-5 mM 4PBA for 2 days in culture demonstrated a dose-dependent reduction in Hsc70 protein immunoreactivity and mRNA levels. Immunoprecipitation with Hsc70-specific antiserum demonstrated that Hsc70 and CFTR associated under control conditions and that treatment with 4PBA reduced these complexes. Levels of immunoreactive Hsp40, Hdj2, Hsp70, Hsp90, and calnexin were unaffected by 4PBA treatment. These data suggest that 4PBA may improve DeltaF508-CFTR trafficking by allowing a greater proportion of mutant CFTR to escape association with Hsc70. PMID- 10666021 TI - Regulation of intestinal vitamin B(2) absorption. Focus on "Riboflavin uptake by human-derived colonic epithelial NCM460 cells". PMID- 10666022 TI - Riboflavin uptake by human-derived colonic epithelial NCM460 cells. AB - Normal microflora of the large intestine synthesize a number of water-soluble vitamins including riboflavin (RF). Recent studies have shown that colonic epithelial cells possess an efficient carrier-mediated mechanism for absorbing some of these micronutrients. The aim of the present study was to determine whether colonic cells also possess a carrier-mediated mechanism for RF uptake and, if so, to characterize this mechanism and study its cellular regulation. Confluent monolayers of the human-derived nontransformed colonic epithelial cells NCM460 and [(3)H]RF were used in the study. Uptake of RF was found to be 1) appreciable and temperature and energy dependent; 2) Na(+) independent; 3) saturable as a function of concentration with an apparent K(m) of 0.14 microM and V(max) of 3.29 pmol x mg protein(-1) x 3 min(-1); 4) inhibited by the structural analogs lumiflavin and lumichrome (K(i) of 1.8 and 14.1 microM, respectively) but not by the unrelated biotin; 5) inhibited in a competitive manner by the membrane transport inhibitor amiloride (K(i) = 0.86 mM) but not by furosemide, DIDS, or probenecid; 6) adaptively regulated by extracellular RF levels with a significant and specific upregulation and downregulation in RF uptake in RF-deficient and oversupplemented conditions, respectively; and 7) modulated by an intracellular Ca(2+)/calmodulin-mediated pathway. These studies demonstrate for the first time the existence of a specialized carrier-mediated mechanism for RF uptake in an in vitro cellular model system of human colonocytes. This mechanism appears to be regulated by extracellular substrate level and by an intracellular Ca(2+)/calmodulin-mediated pathway. It is suggested that the identified transport system may be involved in the absorption of bacterially synthesized RF in the large intestine and that this source of RF may contribute toward RF homeostasis, especially that of colonocytes. PMID- 10666023 TI - Charged residues in the M2 region of alpha-hENaC play a role in channel conductance. AB - The epithelial Na(+) channel (ENaC) is a low-conductance channel that is highly selective for Na(+) and Li(+) over K(+) and impermeable to anions. The molecular basis underlying these conduction properties is not well known. Previous studies with the ENaC subunits demonstrated that the M2 region of alpha-ENaC is critical to channel function. Here we examine the effects of reversing the negative charges of highly conserved amino acids in alpha-subunit human ENaC (alpha-hENaC) M1 and M2 domains. Whole cell and single-channel current measurements indicated that the M2 mutations E568R, E571R, and D575R significantly decreased channel conductance but did not affect Na(+):K(+) permeability. We observed no functional perturbations from the M1 mutation E108R. Whole cell amiloride-sensitive current recorded from oocytes injected with the M2 alpha-hENaC mutants along with wild type (wt) beta- and gamma-hENaC was low (46-93 nA) compared with the wt channel (1-3 microA). To determine whether this reduced macroscopic current resulted from a decreased number of mutant channels at the plasma membrane, we coexpressed mutant alpha-hENaC subunits with green fluorescent protein-tagged beta- and gamma subunits. Confocal laser scanning microscopy of oocytes demonstrated that plasma membrane localization of the mutant channels was the same as that of wt. These experiments demonstrate that acidic residues in the second transmembrane domain of alpha-hENaC affect ion permeation and are thus critical components of the conductive pore of ENaC. PMID- 10666024 TI - Hyperbaric oxygen downregulates ICAM-1 expression induced by hypoxia and hypoglycemia: the role of NOS. AB - Hyperbaric oxygen (HBO) is being studied as a therapeutic intervention for ischemia/reperfusion (I/R) injury. We have developed an in vitro endothelial cell model of I/R injury to study the impact of HBO on the expression of intercellular adhesion molecule-1 (ICAM-1) and polymorphonuclear leukocyte (PMN) adhesion. Human umbilical vein endothelial cell (HUVEC) and bovine aortic endothelial cell (BAEC) induction of ICAM-1 required simultaneous exposure to both hypoxia and hypoglycemia as determined by confocal laser scanning microscopy, ELISA, and Western blot. HBO treatment reduced the expression of ICAM-1 to control levels. Adhesion of PMNs to BAECs was increased following hypoxia/hypoglycemia exposure (3. 4-fold, P < 0.01) and was reduced to control levels with exposure to HBO (P = 0.67). Exposure of HUVECs and BAECs to HBO induced the synthesis of endothelial cell nitric oxide synthase (eNOS). The NOS inhibitor nitro-L-arginine methyl ester attenuated HBO-mediated inhibition of ICAM-1 expression. Our findings suggest that the beneficial effects of HBO in treating I/R injury may be mediated in part by inhibition of ICAM-1 expression through the induction of eNOS. PMID- 10666025 TI - Role of K(+) channel expression in polyamine-dependent intestinal epithelial cell migration. AB - Polyamines are essential for cell migration during early mucosal restitution after wounding in the gastrointestinal tract. Activity of voltage-gated K(+) channels (Kv) controls membrane potential (E(m)) that regulates cytoplasmic free Ca(2+) concentration ([Ca(2+)](cyt)) by governing the driving force for Ca(2+) influx. This study determined whether polyamines are required for the stimulation of cell migration by altering K(+) channel gene expression, E(m), and [Ca(2+)](cyt) in intestinal epithelial cells (IEC-6). The specific inhibitor of polyamine synthesis, alpha-difluoromethylornithine (DFMO, 5 mM), depleted cellular polyamines (putrescine, spermidine, and spermine), selectively inhibited Kv1.1 channel (a delayed-rectifier Kv channel) expression, and resulted in membrane depolarization. Because IEC-6 cells did not express voltage-gated Ca(2+) channels, the depolarized E(m) in DFMO-treated cells decreased [Ca(2+)](cyt) as a result of reduced driving force for Ca(2+) influx through capacitative Ca(2+) entry. Migration was reduced by 80% in the polyamine-deficient cells. Exogenous spermidine not only reversed the effects of DFMO on Kv1.1 channel expression, E(m), and [Ca(2+)](cyt) but also restored cell migration to normal. Removal of extracellular Ca(2+) or blockade of Kv channels (by 4-aminopyridine, 1-5 mM) significantly inhibited normal cell migration and prevented the restoration of cell migration by exogenous spermidine in polyamine-deficient cells. These results suggest that polyamine-dependent intestinal epithelial cell migration may be due partially to an increase of Kv1.1 channel expression. The subsequent membrane hyperpolarization raises [Ca(2+)](cyt) by increasing the driving force (the electrochemical gradient) for Ca(2+) influx and thus stimulates cell migration. PMID- 10666026 TI - Differentiation of human fetal osteoblastic cells and gap junctional intercellular communication. AB - Gap junctional channels facilitate intercellular communication and in doing so may contribute to cellular differentiation. To test this hypothesis, we examined gap junction expression and function in a temperature-sensitive human fetal osteoblastic cell line (hFOB 1.19) that when cultured at 37 degrees C proliferates rapidly but when cultured at 39.5 degrees C proliferates slowly and displays increased alkaline phosphatase activity and osteocalcin synthesis. We found that hFOB 1.19 cells express abundant connexin 43 (Cx43) protein and mRNA. In contrast, Cx45 mRNA was expressed to a lesser degree, and Cx26 and Cx32 mRNA were not detected. Culturing hFOB 1. 19 cells at 39.5 degrees C, relative to 37 degrees C, inhibited proliferation, increased Cx43 mRNA and protein expression, and increased gap junctional intercellular communication (GJIC). Blocking GJIC with 18alpha-glycyrrhetinic acid prevented the increase in alkaline phosphatase activity resulting from culture at 39.5 degrees C but did not affect osteocalcin levels. These results suggest that gap junction function and expression parallel osteoblastic differentiation and contribute to the expression of alkaline phosphatase activity, a marker for fully differentiated osteoblastic cells. PMID- 10666027 TI - Nuclear redistribution of tonicity-responsive enhancer binding protein requires proteasome activity. AB - Tonicity-responsive enhancer binding protein (TonEBP) is the transcription factor that regulates tonicity-responsive expression of the genes for the sodium-myo inositol cotransporter (SMIT) and the sodium-chloride-betaine cotransporter (BGT1). Hypertonicity stimulates the activity of TonEBP due to a combination of increased protein abundance and increased nuclear distribution (proportion of TonEBP that is in the nucleus). We found that inhibitors of proteasome activity markedly reduce the induction of SMIT and BGT1 mRNA in response to hypertonicity. These inhibitors also reduce hypertonicity-induced stimulation of expression of a reporter gene controlled by the tonicity-responsive enhancer. Western and immunohistochemical analyses revealed that the proteasome inhibitors reduce the hypertonicity-induced increase of TonEBP in the nucleus by inhibiting its nuclear redistribution without affecting its abundance. Although the nuclear distribution of TonEBP is sensitive to inhibition of proteasome activity as is that of nuclear factor (NF)-kappaB, the signaling pathways appear to be different in that hypertonicity does not affect the nuclear distribution of NF-kappaB. Conversely, treatment with tumor necrosis factor-alpha increases the nuclear distribution of NF-kappaB but not TonEBP. PMID- 10666028 TI - Inhibition of ornithine decarboxylase induces STAT3 tyrosine phosphorylation and DNA binding in IEC-6 cells. AB - Polyamines are required for the proliferation of the rat intestinal mucosal IEC-6 cell line. Ornithine decarboxylase (ODC) is the enzyme that catalyzes the first step in polyamine synthesis. ODC inhibition not only leads to polyamine depletion but also leads to inhibition of cell proliferation and regulates the expression of the immediate-early genes c-fos, c-myc, and c-jun. Members of the signal transducers and activators of transcription (STAT) transcription factor family bind to the sis-inducible element (SIE) present in the promoters to regulate the expression of a variety of important genes. In the present study, we tested the hypothesis that the STAT3 transcription factor, which is responsible for activation of the acute phase response genes, is activated after inhibition of ODC. We found that inhibition of ODC rapidly induces STAT3 activation as determined by STAT3 tyrosine phosphorylation, translocation of STAT3 from the cytoplasm into the nucleus, and the presence of STAT3 in SIE-dependent DNA protein complexes. STAT3 activation upon inhibition of ODC was accompanied by the activation of a STAT3-dependent reporter construct. Moreover, prolonged polyamine depletion resulted in downregulation of cellular STAT3 levels. PMID- 10666029 TI - Cyclic nucleotide-gated cation channels mediate sodium and calcium influx in rat colon. AB - We found mRNA for the three isoforms of the cyclic nucleotide-gated nonselective cation channel expressed in the mucosal layer of the rat intestine from the duodenum to the colon and in intestinal epithelial cell lines in culture. Because these channels are permeable to sodium and calcium and are stimulated by cGMP or cAMP, we measured 8-bromo-cGMP-stimulated sodium-mediated short-circuit current (I(sc)) in proximal and distal colon and unidirectional (45)Ca(2+) fluxes in proximal colon to determine whether these channels could mediate transepithelial sodium and calcium absorption across the colon. Sodium-mediated I(sc), stimulated by 8-bromo-cGMP, were inhibited by dichlorobenzamil and l-cis-diltiazem, blockers of cyclic nucleotide-gated cation channels, suggesting that these ion channels can mediate transepithelial sodium absorption. Sodium-mediated I(sc) and net transepithelial (45)Ca(2+) absorption were stimulated by heat-stable toxin from Escherichia coli that increases cGMP. Addition of l-cis-diltiazem inhibited the enhanced transepithelial absorption of both ions. These results suggest that cyclic nucleotide-gated cation channels simultaneously increase net sodium and calcium absorption in the colon of the rat. PMID- 10666030 TI - CCK stimulates mob-1 expression and NF-kappaB activation via protein kinase C and intracellular Ca(2+). AB - Supraphysiological concentrations of cholecystokinin (CCK) induce chemokine expression in rat pancreatic acini through the activation of the transcription factor NF-kappaB. In the current study, the intracellular signals involved in these pathophysiological effects of CCK were investigated. CCK induction of mob-1 expression in isolated rat pancreatic acini was blocked by the protein kinase C (PKC) inhibitors GF-109203X and Ro-32-0432 and by the intracellular Ca(2+) chelator BAPTA. CCK induced NF-kappaB nuclear translocation, and DNA binding was also blocked by GF-109203X and BAPTA. Direct activation of PKC with TPA induced mob-1 chemokine expression and activated NF-kappaB DNA binding to a similar extent as did CCK. Increasing intracellular Ca(2+) using ionomycin had no effect on mob-1 mRNA levels or NF-kappaB activity. Both CCK and TPA treatments decreased inhibitory kappaB-alpha (IkappaB-alpha) levels, whereas ionomycin had no effect. However, the effects of TPA on IkappaB-alpha degradation were less complete than for CCK. In combination, TPA and ionomycin degraded IkappaB-alpha to a similar extent as CCK. Therefore, activation of NF-kappaB and mob-1 expression by supraphysiological CCK is likely mediated by both PKC activation and elevated intracellular Ca(2+). PMID- 10666031 TI - Purinergic activation of spontaneous transient outward currents in guinea pig taenia colonic myocytes. AB - Spontaneous transient outward currents (STOCs) were recorded from smooth muscle cells of the guinea pig taenia coli using the whole cell patch-clamp technique. STOCs were resolved at potentials positive to -50 mV. Treating cells with caffeine (1 mM) caused a burst of outward currents followed by inhibition of STOCs. Replacing extracellular Ca(2+) with equimolar Mn(2+) caused STOCs to "run down. " Iberiotoxin (200 nM) or charybdotoxin (ChTX; 200 nM) inhibited large amplitude STOCs, but small-amplitude "mini-STOCs" remained in the presence of these drugs. Mini-STOCs were reduced by apamin (500 nM), an inhibitor of small conductance Ca(2+)-activated K(+) channels (SK channels). Application of ATP or 2 methylthioadenosine 5'-triphosphate (2-MeS-ATP) increased the frequency of STOCs. The effects of 2-MeS-ATP persisted in the presence of charybdotoxin but were blocked by combination of ChTX (200 nM) and apamin (500 nM). 2-MeS-ATP did not increase STOCs in the presence of pyridoxal phosphate 6-azophenyl-2',4' disulfonic acid, a P(2) receptor blocker. Similarly, pretreatment of cells with U 73122 (1 microM), an inhibitor of phospholipase C (PLC), abolished the effects of 2-MeS-ATP. Xestospongin C, an inositol 1,4,5-trisphosphate (IP(3)) receptor blocker, attenuated STOCs, but these events were not affected by ryanodine. The data suggest that purinergic activation through P(2Y) receptors results in localized Ca(2+) release via PLC- and IP(3)-dependent mechanisms. Release of Ca(2+) is coupled to STOCs, which are composed of currents mediated by large conductance Ca(2+)-activated K(+) channels and SK channels. The latter are thought to mediate hyperpolarization and relaxation responses of gastrointestinal muscles to inhibitory purinergic stimulation. PMID- 10666032 TI - Endoplasmic reticulum Ca(2+)-ATPase inhibitors stimulate membrane guanylate cyclase in pancreatic acinar cells. AB - In this study, we show that particulate guanylate cyclase (GC) is present in rat pancreatic acinar cells and is located both on plasma membrane and membranes of endoplasmic reticulum (ER). Western blot analysis indicates that the enzyme isoform GC-A is present in the acinar cell membranes. The specific inhibitors of ER Ca(2+)-ATPase thapsigargin, 2,5-di-(t-butyl)-1,4-hydroquinone (BHQ), and cyclopiazonic acid all activated particulate GC in pancreatic acini, both in membrane fractions and intact cells. These inhibitors also induced dephosphorylation of GC. Dose dependencies of Ca(2+)-ATPase inhibition and GC activation by BHQ are very similar, and those for thapsigargin partially overlap. ER Ca(2+)-ATPase and GC are coimmunoprecipitated both by antisera against membrane GC and by antisera against ER Ca(2+)-ATPase, suggesting a physical association between the two enzymes. The results suggest that thapsigargin and the other inhibitors act through ER Ca(2+)-ATPase to activate membrane GC in pancreatic acinar cells, although their direct effect on GC cannot be excluded. PMID- 10666033 TI - Ras signaling in the inner medullary cell response to urea and NaCl. AB - The small guanine nucleotide-binding protein Ras, activated by peptide mitogens and other stimuli, regulates downstream signaling events to influence transcription. The role of Ras in solute signaling to gene regulation was investigated in the murine inner medullary collecting duct (mIMCD3) cell line. Urea treatment (100-200 mM), but not sham treatment, increased Ras activation 124% at 2 min; the effect of NaCl did not achieve statistical significance. To determine the contribution of Ras activation to urea-inducible signal transduction, mIMCD3 cells were stably transfected with an expression plasmid encoding a dominant negative-acting N17Ras mutant driven by a dexamethasone inducible (murine mammary tumor virus) promoter. After 24 h of induction, selected cell lines exhibited sufficient N17Ras overexpression to abolish epidermal growth factor- and hypotonicity-mediated signaling to extracellular signal-regulated kinase (ERK) phosphorylation, as determined by immunoblotting. Conditional N17Ras overexpression inhibited urea- and NaCl-inducible ERK phosphorylation by 40-50%, but only at 15 min, and not 5 min, of treatment. N17Ras induction, however, almost completely inhibited urea-inducible Egr-1 transcription, as quantitated by luciferase reporter gene assay, but failed to influence tonicity-inducible (TonE-mediated) transcription. N17Ras overexpression also blocked urea-inducible expression of the transcription factor Gadd153 but did not influence osmotic or urea-inducible apoptosis. In addition, urea treatment induced recruitment of the Ras activator Sos to the plasma membrane. Taken together, these observations suggest a role for Ras signaling in the IMCD cell response to urea stress. PMID- 10666034 TI - K-Cl cotransport in vascular smooth muscle and erythrocytes: possible implication in vasodilation. AB - K-Cl cotransport, the electroneutral-coupled movement of K and Cl ions, plays an important role in regulatory volume decrease. We recently reported that nitrite, a nitric oxide derivative possessing potent vasodilation properties, stimulates K Cl cotransport in low-K sheep red blood cells (LK SRBCs). We hypothesized that activation of vascular smooth muscle (VSM) K-Cl cotransport by vasodilators decreases VSM tension. Here we tested this hypothesis by comparing the effects of commonly used vasodilators, hydralazine (HYZ), sodium nitroprusside, isosorbide mononitrate, and pentaerythritol, on K-Cl cotransport in LK SRBCs and in primary cultures of rat VSM cells (VSMCs) and of HYZ-induced K-Cl cotransport activation on relaxation of isolated porcine coronary rings. K-Cl cotransport was measured as the Cl-dependent K efflux or Rb influx in the presence and absence of inhibitors for other K/Rb transport pathways. All vasodilators activated K-Cl cotransport in LK SRBCs and HYZ in VSMCs, and this activation was inhibited by calyculin and genistein, two inhibitors of K-Cl cotransport. KT-5823, a specific inhibitor of protein kinase G, abolished the sodium nitroprusside-stimulated K-Cl cotransport in LK SRBCs, suggesting involvement of the cGMP pathway in K-Cl cotransport activation. Hydralazine, in a dose-dependent manner, and sodium nitroprusside relaxed (independently of the endothelium) precontracted arteries when only K-Cl cotransport was operating and other pathways for K/Rb transport, including the Ca-activated K channel, were inhibited. Our findings suggest that K Cl cotransport may be involved in vasodilation. PMID- 10666035 TI - Smokeless tobacco potentiates VIP-induced DNA synthesis and inactivates NEP 24.11 in oral keratinocytes. AB - The purpose of this study was to determine whether exposure of cultured chemically transformed hamster oral keratinocytes (HCPC-1) to an aqueous extract of smokeless tobacco (STE) potentiates DNA synthesis elicited by vasoactive intestinal peptide (VIP), an autocrine neuropeptide, and, if so, whether this response is associated with inactivation of neutral endopeptidase 24.11 (NEP 24. 11), an ectoenzyme that cleaves and inactivates VIP very effectively, in these cells. I found that STE and VIP each elicited a modest, albeit significant, increase in DNA synthesis in cultured HCPC-1 cells (P < 0.05). However, incubation of HCPC-1 cells with STE together with VIP evoked a significant, concentration- dependent increase in DNA synthesis that was mediated by VIP receptors. The effects of STE and VIP were synergistic. Maximal response was observed after a 48-h incubation. STE significantly attenuated NEP 24.11 activity in HCPC-1 cells at a time when VIP-induced DNA synthesis was maximal. Collectively, these data indicate that STE potentiates VIP-induced DNA synthesis in cultured oral keratinocytes, and that this response is temporally related to STE-induced inactivation of NEP 24.11 in these cells. I suggest that NEP 24.11 modulates the mitogenic effects of smokeless tobacco in the oral epithelium, in part, by inactivating VIP. PMID- 10666036 TI - RPTPmu and protein tyrosine phosphorylation regulate K(+) channel mRNA expression in adult cardiac myocytes. AB - Previously, we reported that cell-cell contact regulates K(+) channel mRNA expression in cultured adult rat cardiac myocytes. Here we show that exposing cardiac myocytes to tyrosine kinase inhibitors (genistein, tyrphostin A25), but not inactive analogs, prevents downregulation of Kv1.5 mRNA and upregulation of Kv4.2 mRNA normally observed when they are cultured under low-density conditions. Furthermore, cardiac myocytes cocultured with cells that endogenously (Mv 1 Lu) or heterologously (Chinese hamster ovary cells) express the receptor-type protein tyrosine phosphatase mu (RPTPmu) display Kv1.5 mRNA levels paralleling that which was observed in myocytes cultured under high-density conditions and in intact tissue. In contrast, myocytes cocultured with control cells failed to produce this response. Finally, it is shown that Kv4.2 mRNA expression is unaffected by RPTPmu. These findings reveal that multiple tyrosine phosphorylation-dependent mechanisms control cardiac myocyte K(+) channel genes. Furthermore, we conclude that RPTPmu specifically regulates cardiac myocyte Kv1.5 mRNA expression. Thus this receptor protein tyrosine phosphatase may be important in responses to pathological conditions associated with the loss of cell-cell interactions in the heart. PMID- 10666037 TI - Pinacidil suppresses contractility and preserves energy but glibenclamide has no effect during muscle fatigue. AB - The effects of 10 microM glibenclamide, an ATP-sensitive K(+) (K(ATP)) channel blocker, and 100 microM pinacidil, a channel opener, were studied to determine how the K(ATP) channel affects mouse extensor digitorum longus (EDL) and soleus muscle during fatigue. Fatigue was elicited with 200-ms-long tetanic contractions every second. Glibenclamide did not affect rate and extent of fatigue, force recovery, or (86)Rb(+) fractional loss. The only effects of glibenclamide during fatigue were: an increase in resting tension (EDL and soleus), a depolarization of the cell membrane, a prolongation of the repolarization phase of action potential, and a greater ATP depletion in soleus. Pinacidil, on the other hand, increased the rate but not the extent of fatigue, abolished the normal increase in resting tension during fatigue, enhanced force recovery, and increased (86)Rb(+) fractional loss in both the EDL and soleus. During fatigue, the decreases in ATP and phosphocreatine of soleus muscle were less in the presence of pinacidil. The glibenclamide effects suggest that fatigue, elicited with intermittent contractions, activates few K(ATP) channels that affect resting tension and membrane potentials but not tetanic force, whereas opening the channel with pinacidil causes a faster decrease in tetanic force, improves force recovery, and helps in preserving energy. PMID- 10666038 TI - PKA holoenzyme is functionally coupled to CFTR by AKAPs. AB - Cystic fibrosis transmembrane regulator (CFTR) is reported to be preferentially regulated by membrane-bound protein kinase A (PKAII). We tested for close physical and functional association of PKA with CFTR in inside-out membrane patches excised from Calu-3 cells. In the presence of MgATP, 8-(4 chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT-cAMP) increased the product of CFTR channel number and open probability (from 0.36 +/- 0.12 to 1.23 +/- 0.57, n = 20, P < 0.0025), and this stimulation was abolished by PKI. Thus Calu-3 membrane isolated from cells retains PKA holoenzyme that is functionally coupled to CFTR. PKAII is anchored at specific subcellular sites by A kinase anchoring proteins (AKAPs). Exposure of excised patches to HT-31, a peptide that disrupts the association of PKAII and AKAPs, prevented CPT-cAMP stimulation of CFTR. Therefore, PKA holoenzyme in isolated membrane patches is bound to AKAPs. In whole cell voltage-clamp studies, intracellular dialysis of Calu-3 cells with HT-31 blocked the activation of CFTR by extracellular adenosine. These results suggest that AKAPs mediate PKA compartmentalization with CFTR and are required for activation of CFTR by physiological regulators. PMID- 10666039 TI - Ca(2+) activation of heart mitochondrial oxidative phosphorylation: role of the F(0)/F(1)-ATPase. AB - Ca(2+) has been postulated as a cytosolic second messenger in the regulation of cardiac oxidative phosphorylation. This hypothesis draws support from the well known effects of Ca(2+) on muscle activity, which is stimulated in parallel with the Ca(2+)-sensitive dehydrogenases (CaDH). The effects of Ca(2+) on oxidative phosphorylation were further investigated in isolated porcine heart mitochondria at the level of metabolic driving force (NADH or Deltapsi) and ATP production rates (flow). The resulting force-flow (F-F) relationships permitted the analysis of Ca(2+) effects on several putative control points within oxidative phosphorylation, simultaneously. The F-F relationships resulting from additions of carbon substrates alone provided a model of pure CaDH activation. Comparing this curve with variable Ca(2+) concentration ([Ca(2+)]) effects revealed an approximate twofold higher ATP production rate than could be explained by a simple increase in NADH or Deltapsi via CaDH activation. The half-maximal effect of Ca(2+ )at state 3 was 157 nM and was completely inhibited by ruthenium red (1 microM), indicating matrix dependence of the Ca(2+) effect. Arsenate was used as a probe to differentiate between F(0)/F(1)-ATPase and adenylate translocase activity by a futile recycling of ADP-arsenate within the matrix, catalyzed by the F(0)/F(1)-ATPase. Ca(2+) increased the ADP arsenylation rate more than twofold, suggesting a direct effect on the F(0)/F(1)-ATPase. These results suggest that Ca(2+) activates cardiac aerobic respiration at the level of both the CaDH and F(0)/F(1)-ATPase. This type of parallel control of both intermediary metabolism and ATP synthesis may provide a mechanism of altering ATP production rates with minimal changes in the high-energy intermediates as observed in vivo. PMID- 10666040 TI - cAMP-activated anion conductance is associated with expression of CFTR in neonatal mouse cardiac myocytes. AB - In this study, patch-clamp techniques were applied to cultured neonatal mouse cardiac myocytes (NMCM) to assess the contribution of cAMP stimulation to the anion permeability in this cell model. Addition of either isoproterenol or a cocktail to raise intracellular cAMP increased the whole cell currents of NMCM. The cAMP-dependent conductance was largely anionic, as determined under asymmetrical (low intracellular) Cl(-) conditions and symmetrical Cl(-) in the presence of various counterions, including Na(+), Mg(2+), Cs(+), and N-methyl-D glucamine. Furthermore, the cAMP-stimulated conductance was also permeable to ATP. The cAMP-activated currents were inhibited by diphenylamine-2-carboxylate, glibenclamide, and an anti-cystic fibrosis transmembrane conductance regulator (CFTR) monoclonal antibody. The anti-CFTR monoclonal antibody failed, however, to inhibit an osmotically activated anion conductance, indicating that CFTR is not linked to osmotically stimulated currents in this cell model. Immunodetection studies of both neonatal mouse heart tissue and cultured NMCM revealed that CFTR is expressed in these preparations. The implication of CFTR in the cAMP stimulated Cl(-)- and ATP-permeable conductance was further verified with NMCM of CFTR knockout mice [cftr(-/-)] in which cAMP stimulation was without effect on the whole cell currents. In addition, stimulation with protein kinase A and ATP induced Cl(-)-permeable single-channel activity in excised, inside-out patches from control, but not cftr(-/-) NMCM. The data in this report indicate that cAMP stimulation of NMCM activates an anion-permeable conductance with functional properties similar to those expected for CFTR, thus suggesting that CFTR may be responsible for the cAMP-activated conductance. CFTR may thus contribute to the permeation and/or regulation of Cl(-)- and ATP-permeable pathways in the developing heart. PMID- 10666041 TI - Lessons from genetically engineered animal models VIII. Absorption and secretion of ions in the gastrointestinal tract. AB - Absorption and secretion of ions in gastrointestinal and other epithelial tissues require the concerted activities of ion pumps, channels, symporters, and exchangers, which operate in coupled systems to mediate transepithelial transport. Our understanding of the identities, membrane locations, and biochemical activities of epithelial ion transporters has advanced significantly in recent years, but major gaps and uncertainties remain in our understanding of their physiological functions. Increasingly, this problem is being addressed by the analysis of mutant mouse models developed by gene targeting. In this review, we discuss gene knockout studies of the secretory isoform of the Na(+)-K(+)-2Cl( ) cotransporter, isoforms 1, 2, and 3 of the Na(+)/H(+) exchanger, and the colonic H(+)-K(+)-ATPase. This approach is leading to a clearer understanding of the functions of these transporters in the living animal. PMID- 10666042 TI - Nutrient tasting and signaling mechanisms in the gut V. Mechanisms of immunologic sensation of intestinal contents. AB - Immune perception of intestinal contents reflects a functional dualism with systemic hyporesponsiveness to dietary antigens and resident microflora (oral tolerance) and active immune responses to mucosal pathogens. This facilitates optimal absorption of dietary nutrients while conserving immunologic resources for episodic pathogenic challenge. Discrimination between dangerous and harmless antigens within the enteric lumen requires continual sampling of the microenvironment by multiple potential pathways, innate and adaptive recognition mechanisms, bidirectional lymphoepithelial signaling, and rigorous control of effector responses. Errors in these processes disrupt mucosal homeostasis and are associated with food hypersensitivity and mucosal inflammation. Mechanisms of mucosal immune perception and handling of dietary proteins and other antigens have several practical and theoretical implications including vaccine design, therapy of systemic autoimmunity, and alteration of enteric flora with probiotics. PMID- 10666043 TI - NHE1, NHE2, and NHE3 contribute to regulation of intracellular pH in murine duodenal epithelial cells. AB - Na(+)/H(+)-exchangers (NHE) mediate acid extrusion from duodenal epithelial cells, but the isoforms involved have not previously been determined. Thus we investigated 1) the contribution of Na(+)-dependent processes to acid extrusion, 2) sensitivity to Na(+)/H(+) exchange inhibitors, and 3) molecular expression of NHE isoforms. By fluorescence spectroscopy the recovery of intracellular pH (pH(i)) was measured on suspensions of isolated acidified murine duodenal epithelial cells loaded with 2', 7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. Expression of NHE isoforms was studied by RT-PCR and Western blot analysis. Reduction of extracellular Na(+) concentration ([Na(+)](o)) during pH(i) recovery decreased H(+) efflux to minimally 12.5% of control with a relatively high apparent Michaelis constant for extracellular Na(+). The Na(+)/H(+) exchange inhibitors ethylisopropylamiloride and amiloride inhibited H(+) efflux maximally by 57 and 80%, respectively. NHE1, NHE2, and NHE3 were expressed at the mRNA level (RT-PCR) as well as at the protein level (Western blot analysis). On the basis of the effects of low [Na(+)](o) and inhibitors we propose that acid extrusion in duodenal epithelial cells involves Na(+)/H(+) exchange by isoforms NHE1, NHE2, and NHE3. PMID- 10666044 TI - Genistein augments prostaglandin-induced recovery of barrier function in ischemia injured porcine ileum. AB - We have previously shown that PGE(2) enhances recovery of transmucosal resistance (R) in ischemia-injured porcine ileum via a mechanism involving chloride secretion. Because the tyrosine kinase inhibitor genistein amplifies cAMP-induced Cl(-) secretion, we postulated that genistein would augment PGE(2)-induced recovery of R. Porcine ileum subjected to 45 min of ischemia was mounted in Ussing chambers, and R and mucosal-to-serosal fluxes of [(3)H]N-formyl-methionyl leucyl phenylalanine (FMLP) and [(3)H]mannitol were monitored as indicators of recovery of barrier function. Treatment with genistein (10(-4) M) and PGE(2) (10( 6) M) resulted in synergistic elevations in R and additive reductions in mucosal to-serosal fluxes of [(3)H]FMLP and [(3)H]mannitol, whereas treatment with genistein alone had no effect. Treatment of injured tissues with genistein and either 8-bromo-cAMP (10(-4) M) or cGMP (10(-4) M) resulted in synergistic increases in R. However, treatment of tissues with genistein and the protein kinase C (PKC) agonist phorbol myristate acetate (10(-5)-10(-6) M) had no effect on R. Genistein augments recovery of R in the presence of cAMP or cGMP but not in the presence of PKC agonists. PMID- 10666045 TI - Glucocorticoids and dietary iron regulate postnatal intestinal heavy and light ferritin expression in rats. AB - To cope with increasing dietary iron exposure, the intestinal epithelium of weaning rats must control intracellular labile iron pools. Intestinal expression of heavy (H) and light (L) ferritin subunits during early weaning and after cortisone administration and/or iron feeding was investigated. Changes in H and L ferritin gene expression were determined by nuclear runoff transcriptional assay, Northern blot analysis, and metabolic labeling of protein synthesis. H ferritin mRNA levels did not change between days 12 and 15, doubled on day 18, and tripled on day 24. L ferritin mRNA was reduced by 50% on days 18 and 24. The protein level of the H and L subunits paralleled the change in mRNAs. Cortisone treatment on day 12 induced a precocious increase of H and decrease of L mRNA expression on day 15. Nuclear runoff assays showed that cortisone did not change H and reduced L ferritin gene transcription. The increased level of H mRNA by cortisone was not translated, unless the rats were fed an iron-fortified diet, which reduced iron regulatory protein activity and stimulated a three- to sixfold increase of ferritin synthesis. Thus changes in intestinal H and L ferritin expression in weaning rats are modulated by glucocorticoids and iron; the former stabilizes H mRNA and suppresses L ferritin gene transcription, and the latter derepresses translation of ferritin mRNA. PMID- 10666046 TI - Release of osmolytes induced by phagocytosis and hormones in rat liver. AB - Betaine, taurine, and inositol participate as osmolytes in liver cell volume homeostasis and interfere with cell function. In this study we investigated whether osmolytes are also released from the intact liver independent of osmolarity changes. In the perfused rat liver, phagocytosis of carbon particles led to a four- to fivefold stimulation of taurine efflux into the effluent perfusate above basal release rates. This taurine release was inhibited by 70-80% by the anion exchange inhibitor DIDS or by pretreatment of the rats with gadolinium chloride. Administration of vasopressin, cAMP, extracellular ATP, and glucagon also increased release of betaine and/or taurine, whereas insulin, extracellular UTP, and adenosine were without effect. In isolated liver cells, it was shown that parenchymal cells and sinusoidal endothelial cells, but not Kupffer cells and hepatic stellate cells, release osmolytes upon hormone stimulation. This may be caused by a lack of hormone receptor expression in these cells, because single-cell fluorescence measurements revealed an increase of intracellular calcium concentration in response to vasopressin and glucagon in parenchymal cells and sinusoidal endothelial cells but not in Kupffer cells and hepatic stellate cells. The data show that Kupffer cells release osmolytes during phagocytosis via DIDS-sensitive anion channels. This mechanism may be used to compensate for the increase in cell volume induced by the ingestion of phagocytosable material. The physiological significance of hormone-induced osmolyte release remains to be evaluated. PMID- 10666047 TI - Impairment of Ca(2+) mobilization in circular muscle cells of the inflamed colon. AB - This study investigated whether inflammation modulates the mobilization of Ca(2+) in canine colonic circular muscle cells. The contractile response of single cells from the inflamed colon was significantly suppressed in response to ACh, KCl, and BAY K8644. Methoxyverapamil and reduction in extracellular Ca(2+) concentration dose-dependently blocked the response in both normal and inflamed cells. The increase in intracellular Ca(2+) concentration in response to ACh and KCl was significantly reduced in the inflamed cells. However, Ca(2+) efflux from the ryanodine- and inositol 1,4, 5-trisphosphate (IP(3))-sensitive stores, as well as the decrease of cell length in response to ryanodine and IP(3), were not affected. Heparin significantly blocked Ca(2+) efflux and contraction in response to ACh in both conditions. ACh-stimulated accumulation of IP(3) and the binding of [(3)H]ryanodine to its receptors were not altered by inflammation. Ruthenium red partially inhibited the response to ACh in normal and inflamed states. We conclude that the canine colonic circular muscle cells utilize Ca(2+) influx through L-type channels as well as Ca(2+) release from the ryanodine- and IP(3) sensitive stores to contract. Inflammation impairs Ca(2+) influx through L-type channels, but it may not affect intracellular Ca(2+) release. The impairment of Ca(2+) influx may contribute to the suppression of circular muscle contractility in the inflamed state. PMID- 10666048 TI - Ionizing radiation induces iNOS-mediated epithelial dysfunction in the absence of an inflammatory response. AB - Ionizing radiation induces intestinal epithelial hyporesponsiveness to secretagogues through an unknown mechanism. We investigated the role of the inducible isoform of nitric oxide (NO) synthase (iNOS)-derived NO in radiation induced hyporesponsiveness. C57BL/6 mice were sham treated or exposed to 10-Gy gamma-radiation and were studied 3 days later. Tissues were mounted in Ussing type diffusion chambers to assess chloride secretion in response to electrical field stimulation (EFS) and forskolin (10 microM). Transport studies were also repeated in iNOS-deficient mice. White blood cell counts were significantly lower in irradiated mice, and there was no inflammatory response as shown by myeloperoxidase activity and histological assessment. iNOS mRNA levels and nitrate/nitrite concentrations were significantly elevated in irradiated colons. iNOS immunoreactivity localized to the epithelium. Colons from irradiated wild type, but not iNOS-deficient, mice exhibited a significant reduction in the responsiveness of the tissue to EFS and forskolin. The hyporesponsiveness was reversed by L-N(6)-(1-iminoethyl)lysine, 1400W, and dexamethasone treatments. iNOS-derived NO mediates colonic hyporesponsiveness 3 days after irradiation in the mouse in the absence of an inflammatory response. PMID- 10666049 TI - Defect of receptor-G protein coupling in human gallbladder with cholesterol stones. AB - Human gallbladders with cholesterol stones (ChS) exhibit an impaired muscle contraction and relaxation and a lower CCK receptor-binding capacity compared with those with pigment stones (PS). This study was designed to determine whether there is an abnormal receptor-G protein coupling in human gallbladders with ChS using (35)S-labeled guanosine 5'-O-(3-thiotriphosphate) ([(35)S]GTPgammaS) binding, (125)I-labeled CCK-8 autoradiography, immunoblotting, and G protein quantitation. CCK and vasoactive intestinal peptide caused significant increases in [(35)S]GTPgammaS binding to Galpha(i-3) and G(s)alpha, respectively. The binding was lower in ChS than in PS (P < 0.01). The reduced [(35)S]GTPgammaS binding in ChS was normalized after the muscles were treated with cholesterol free liposomes (P < 0.01). Autoradiography and immunoblots showed a decreased optical density (OD) for CCK receptors, an even lower OD value for receptor-G protein coupling, and a higher OD for uncoupled receptors or Galpha(i-3) protein in ChS compared with PS (P < 0.001). G protein quantitation also showed that there were no significant differences in the Galpha(i-3) and G(s)alpha content in ChS and PS. We conclude that, in addition to an impaired CCK receptor-binding capacity, there is a defect in receptor-G protein coupling in muscle cells from gallbladder with ChS. These changes may be normalized after removal of excess cholesterol from the plasma membrane. PMID- 10666050 TI - Inflammation-induced impairment of enteric nerve function in nematode-infected mice is macrophage dependent. AB - Trichinella spiralis infection in rodents is associated with suppression of ACh release from myenteric plexus that can be mimicked by macrophage-derived cytokines. We verified the presence of a macrophage infiltrate in the intestine during T. spiralis infection and determined the extent to which this cell type is responsible for the neural changes. C57BL/6 mice were infected with 375 T. spiralis larvae by gavage, and the presence of macrophages (F4/80 positive) in the jejunum was determined immunohistochemically. In another experiment, infected mice were treated intravenously with liposomes containing dichloromethylene diphosphonate (clodronate, Cl(2)MDP), which causes apoptosis of macrophages, and killed at postinfection day 6, and jejunal tissues were evaluated for the presence of F4/80-positive cells and for [(3)H]ACh release from the myenteric plexus. Infection caused an infiltration of F4/80-positive cells into the intestinal mucosa, muscle layers, and myenteric plexus region and a significant suppression of ACh release (50%). Depletion of F4/80-positive macrophages using Cl(2)MDP-containing liposomes prevented the suppression in [(3)H]ACh release, identifying macrophages as the cell type involved in the functional impairment of enteric cholinergic nerves. PMID- 10666051 TI - Inhibition of rat colon tumors by sulindac and sulindac sulfone is independent of K-ras (codon 12) mutation. AB - Nonsteroidal anti-inflammatory drug (NSAID) use reduces the risk of colorectal cancer by 40-50%. Previous studies suggest that effective inhibition of colorectal cancer by NSAIDs may be dependent on the presence or absence of a K ras mutation. This study was aimed at determining the relationship between inhibition of colorectal cancer by sulindac and sulindac sulfone and the presence of activating K-ras mutations in the 1,2-dimethylhydrazine dihydrochloride rat model. Sulindac (20 mg x kg(-1) x day(-1)), sulindac sulfone (40 mg x kg(-1) x day(-1)), or vehicle was administered orally to male Sprague-Dawley rats for a 4 wk period beginning 20 wk after tumor induction. Tumor number and volume were measured before treatment by laparotomy and colonoscopy and again after treatment. Sulindac and sulindac sulfone treatment significantly reduced the number and volume of colorectal tumors compared with control rats. For K-ras (codon 12) mutation detection, frozen tumor tissue was collected at the endpoint. We found K-ras codon 12 mutations in 11 of 21 (52%) control tumors. The proportion of tumors with K-ras mutations in the sulindac-treated group [5 of 8 (62%); odds ratio = 1.51 (95% confidence interval = 0.29, 8.33)] and the proportion of sulindac sulfone-treated tumors [9 of 14 (64%); odds ratio = 1.63 (95% confidence interval = 0.41, 6.66)] were not significantly different from controls. Tumor inhibition did not correlate with K-ras (codon 12) mutation status, which suggests that the mechanism of inhibition of rat colorectal cancer by sulindac and sulindac sulfone is independent of K-ras mutation. PMID- 10666052 TI - Properties of synaptic inputs from myenteric neurons innervating submucosal S neurons in guinea pig ileum. AB - This study examined synaptic inputs from myenteric neurons innervating submucosal neurons. Intracellular recordings were obtained from submucosal S neurons in guinea pig ileal preparations in vitro, and synaptic inputs were recorded in response to electrical stimulation of exposed myenteric plexus. Most S neurons received synaptic inputs [>80% fast (f) excitatory postsynaptic potentials (EPSP), >30% slow (s) EPSPs] from the myenteric plexus. Synaptic potentials were recorded significant distances aboral (fEPSPs, 25 mm; sEPSPs, 10 mm) but not oral to the stimulating site. When preparations were studied in a double-chamber bath that chemically isolated the stimulating "myenteric chamber" from the recording side "submucosal chamber," all fEPSPs were blocked by hexamethonium in the submucosal chamber, but not by a combination of nicotinic, purinergic, and 5 hydroxytryptamine-3 receptor antagonists in the myenteric chamber. In 15% of cells, a stimulus train elicited prolonged bursts of fEPSPs (>30 s duration) that were blocked by hexamethonium. These findings suggest that most submucosal S neurons receive synaptic inputs from predominantly anally projecting myenteric neurons. These inputs are poised to coordinate intestinal motility and secretion. PMID- 10666053 TI - Evidence for a peripheral mechanism of esophagocrural diaphragm inhibitory reflex in cats. AB - The esophagogastric junction (EGJ) is guarded by two sphincters, a smooth muscle lower esophageal sphincter (LES) and a skeletal muscle crural diaphragm. These two sphincters relax simultaneously under certain physiological conditions, i.e., swallowing, belching, vomiting, transient LES relaxation, and esophageal distension. Esophageal distension-induced crural diaphragm relaxation is mediated through vagal afferents that are thought to exert inhibitory influence on the central mechanism (brain stem) of crural diaphragm contraction. We conducted studies in 10 cats to determine whether a mechanism of crural diaphragm relaxation was located at the level of the neuromuscular junction and/or muscle. Stimulation of the crural diaphragm neuromuscular junction through 1) the electrodes implanted in the muscle and 2) the bilateral phrenic nerve resulted in an increase in EGJ pressure. Nicotinic receptor blockade (pancuronium, 0.2 mg/kg) abolished the EGJ pressure increase caused by electrical stimulation of the neuromuscular junction. Esophageal distension and bolus-induced secondary esophageal peristalsis caused relaxation of the EGJ during the stimulation of the neuromuscular junction. Bilateral phrenicotomy and vagotomy had no influence on this relaxation. These data suggest the existence of a peripheral mechanism of crural diaphragm inhibition. This peripheral inhibitory mechanism may reside at the level of either the neuromuscular junction or the skeletal muscle. PMID- 10666054 TI - Kir3.1/3.2 encodes an I(KACh)-like current in gastrointestinal myocytes. AB - Expression of the Kir3 channel subfamily in gastrointestinal (GI) myocytes was investigated. Members of this K(+) channel subfamily encode G protein-gated inwardly rectifying K(+) channels (I(KACh)) in other tissues, including the heart and brain. In the GI tract, I(KACh) could act as a negative feedback mechanism to temper the muscarinic response mediated primarily through activation of nonselective cation currents and inhibition of delayed-rectifier conductance. Kir3 channel subfamily isoforms expressed in GI myocytes were determined by performing RT-PCR on RNA isolated from canine colon, ileum, duodenum, and jejunum circular myocytes. Qualitative PCR demonstrated the presence of Kir3.1 and Kir3.2 transcripts in all smooth muscle cell preparations examined. Transcripts for Kir3.3 and Kir3.4 were not detected in the same preparations. Semiquantitative PCR showed similar transcriptional levels of Kir3.1 and Kir3.2 relative to beta actin expression in the various GI preparations. Full-length cDNAs for Kir3.1 and Kir3.2 were cloned from murine colonic smooth muscle RNA and coexpressed in Xenopus oocytes with human muscarinic type 2 receptor. Superfusion of oocytes with ACh (10 microM) reversibly activated a Ba(2+)-sensitive and inwardly rectifying K(+) current. Immunohistochemistry using Kir3.1- and Kir3.2-specific antibodies demonstrated channel expression in circular and longitudinal smooth muscle cells. We conclude that an I(KACh) current is expressed in GI myocytes encoded by Kir3.1/3.2 heterotetramers. PMID- 10666055 TI - Propagation of individual spikes as "patches" of activation in isolated feline duodenum. AB - Asynchrony of spikes has made it difficult to study the spatial and temporal behavior of spikes in the gastrointestinal system. By simultaneously recording from a large number of closely spaced electrodes, we investigated the propagation of individual spikes. Recordings were performed from the serosal surface of the isolated feline duodenum at 240 sites simultaneously. Analysis of the tracings made it possible to reconstruct the propagation of individual spikes. Spikes propagate in the longitudinal and circumferential directions in self-limiting areas or "patches." Conduction within patches may occur in the orad or aborad direction irrespective of the direction of the slow wave. Most of the patches are smaller (<40 mm(2)), although inhomogeneous activation by the preceding slow wave may increase their size. Stimulation by ACh, TTX, or tetraethylammonium does not affect the average patch size but does increase significantly their number and distribution in the duodenum [from 26% (control) to 56%, 61%, and 72%, respectively]. In conclusion, individual spikes activate limited areas or patches in the small intestine, and pharmacological stimulation increases the number and distribution of these patches. In the small intestine, this pattern of activation would induce localized contractions. Contraction could be modulated by the size, number, and distribution of spike patches. PMID- 10666056 TI - Syncollin is differentially expressed in rat proximal small intestine and regulated by feeding behavior. AB - Gradients of gene expression are maintained along the proximal-distal axis of the mammalian small intestine despite a continuously regenerating epithelium. To study the molecular mechanisms responsible for this phenomenon, we utilized a subtractive hybridization strategy to isolate genes differentially expressed in the duodenum but not ileum. We isolated and sequenced 15 clones. The clones were fragments of genes encoding lipases, proteases, and an esterase. A novel clone was characterized and subsequently shown to encode syncollin, a secretory granule protein that binds to syntaxin in a calcium-sensitive manner. RT-PCR and S1 nuclease protection assay were used to clarify the 5'-end of syncollin. Syncollin was expressed in the rat pancreas, spleen, duodenum, and colon. In situ hybridization localized syncollin expression in the pancreas to acinar cells and in the duodenum to villus epithelial cells. PMID- 10666057 TI - c-Myb modulates transcription of the alpha-smooth muscle actin gene in activated hepatic stellate cells. AB - Expression of alpha-smooth muscle actin (alpha-SMA) defines the phenotype of activated (myofibroblastic) hepatic stellate cells. These cells, but not quiescent stellate cells, have a high level of alpha-SMA and c-Myb expression, as well as increased c-Myb-binding activities to the proximal alpha-SMA E box. Therefore, we analyzed the role of c-Myb in alpha-SMA transcription and stellate cell activation. Activated primary rat stellate cells displayed a high expression of the -724 and -271 alpha-SMA/luciferase (LUC) chimeric genes, which contain c Myb binding sites (-223/-216 bp). Alpha-SMA/LUC minigenes with mutation (-219/ 217 bp), truncation (-224 bp), or deletion (-191 bp) of the c-Myb binding site were not efficiently transcribed. Transfection of wild-type c-Myb into quiescent stellate cells, which do not express endogenous c-Myb, induced a approximately 10 fold stimulation of -724 alpha-SMA/LUC expression. Conversely, expression of either a dominant-negative c-Myb basic domain mutant (Cys(43) --> Asp) or a c-Myb antisense RNA blocked transcription from the -724 alpha-SMA/LUC or -271 alpha SMA/LUC in activated cells. Moreover, transfection of c-myb antisense, but not sense, RNA inhibited both expression of the endogenous alpha-SMA gene and stellate cell activation, whereas transfection of c-myb stimulated alpha-SMA expression in quiescent stellate cells. These findings suggest that c-Myb modulates the activation of stellate cells and that integrity of the redox sensor Cys(43) in c-Myb is required for this effect. PMID- 10666058 TI - Relationships between spatial patterns of colonic pressure and individual movements of content. AB - The aim of this study was to examine the relationship between colonic pressure waves and movement of content. In 11 healthy subjects, pressures were recorded at 10-cm intervals from cecum to rectum for 32 h. In six subjects, transit was simultaneously measured for 8 h after direct cecal instillation of 1.5 mCi of (99m)Tc sulfur colloid. Thirty-two percent of isotope movements were related to nonpropagating activity and twenty-eight percent to propagating sequences. The extent of isotope movement related to propagating sequences (25.1 +/- 2.1 cm) was greater than that due to nonpropagating activity (12.8 +/- 0.7 cm; P = 0.0001). Propagating sequences originated significantly more frequently (P = 0.004) and propagated further (P = 0.0006) in the proximal compared with the distal colon. Only 36% of propagating sequences were propulsive of content, and compared with nonpropulsive sequences, these propagated further (41 +/- 6 vs. 27 +/- 2 cm; P < 0.05) and had a higher probability of originating proximally (P = 0.0003), a higher pressure wave amplitude (50 +/- 5 vs. 34 +/- 4 mmHg; P = 0.0001), and slower velocity (2.2 +/- 0.3 vs. 3.6 +/- 0.47 cm/s; P = 0.02). We conclude that most movements of colonic content are related to pressure waves. There is marked regional variation in the prevalence, velocity, and extent of propagation of propagating pressure wave sequences, which are an important mechanism for transporting content over long distances. The effectiveness of transport by a propagating sequence is influenced by its site of origin, amplitude, and velocity. PMID- 10666059 TI - Mechanism of preserved positive lusitropy by cAMP-dependent drugs in heart failure. AB - In tachycardia-induced heart failure (HF), positive lusitropic effects of milrinone or dobutamine were assessed by evaluating the time constant of left ventricular (LV) pressure decay (tau) and Ca(2+)-ATPase activity of the sarcoplasmic reticulum (SR). The peak value of the positive first derivative of LV pressure (+dP/dt) was less increased, either by dobutamine (2-10 microg x kg( 1) x min(-1)) or by milrinone (4-20 microg/kg), in HF than in control (P < 0.05), whereas tau was shortened to an extent similar to that in control with dobutamine [P = not significant (NS)] and to an even greater extent with milrinone (P < 0.05). Ca(2+)-ATPase activity increased similarly in HF and control with dobutamine (1 microM; +11% in HF vs. +12% in control, P = NS), whereas it increased more with milrinone (1 microM; +19% in HF vs. +11% in control, P < 0.05). Ca(2+)-ATPase activity-cAMP relationships were shifted to the left by milrinone or dobutamine in HF compared with control. Thus, in HF, the sensitivity of Ca(2+)-ATPase activity to cAMP was increased on addition of cAMP-dependent inotropic agents, contributing to the preservation of positive lusitropy. PMID- 10666060 TI - Stroke volume during exercise: interaction of environment and hydration. AB - Euhydrated and dehydrated subjects exercised in a hot and a cold environment with our aim to identify factors that relate to reductions in stroke volume (SV). We hypothesized that reductions in SV with heat stress are related to the interaction of several factors rather than the effect of elevated skin blood flow. Eight male endurance-trained cyclists [maximal O(2) consumption (VO(2 max)) 4.5 +/- 0.1 l/min; means +/- SE] cycled for 30 min (72% VO(2 max)) in the heat (H; 35 degrees C) or the cold (C; 8 degrees C) when euhydrated or dehydrated by 1.5, 3.0, or 4.2% of their body weight. When euhydrated, SV and esophageal temperature (T(es) 38. 2-38.3 degrees C) were similar in H and C, whereas skin blood flow was much higher in H vs. C (365 +/- 64% higher; P < 0.05). With each 1% body weight loss, SV declined 6.4 +/- 1.3 ml (4.8%) in H and 3.4 +/- 0.4 ml (2.5%) in C, whereas T(es) increased 0.21 +/- 0.02 and 0. 10 +/- 0.02 degrees C in H and C, respectively (P < 0.05). However, reductions in SV were not associated with increases in skin blood flow. The reduced SV was highly associated with increased heart rate and reduced blood volume in both H (R = 0.96; P < 0.01) and C (R = 0. 85; P < 0.01). In conclusion, these results suggest that SV is maintained in trained subjects during exercise in euhydrated conditions despite large differences in skin blood flow. Furthermore, the lowering of SV with dehydration appears largely related to increases in heart rate and reductions in blood volume. PMID- 10666061 TI - Chronic hypercapnia inhibits hypoxic pulmonary vascular remodeling. AB - Chronic hypercapnia is commonly found in patients with severe hypoxic lung disease and is associated with a greater elevation of pulmonary arterial pressure than that due to hypoxia alone. We hypothesized that hypercapnia worsens hypoxic pulmonary hypertension by augmenting pulmonary vascular remodeling and hypoxic pulmonary vasoconstriction (HPV). Rats were exposed to chronic hypoxia [inspiratory O(2) fraction (FI(O(2))) = 0.10], chronic hypercapnia (inspiratory CO(2) fraction = 0.10), hypoxia-hypercapnia (FI(O(2)) = 0.10, inspiratory CO(2) fraction = 0.10), or room air. After 1 and 3 wk of exposure, muscularization of resistance blood vessels and hypoxia-induced hematocrit elevation were significantly inhibited in hypoxia-hypercapnia compared with hypoxia alone (P < 0.001, ANOVA). Right ventricular hypertrophy was reduced in hypoxia-hypercapnia compared with hypoxia at 3 wk (P < 0.001, ANOVA). In isolated, ventilated, blood perfused lungs, basal pulmonary arterial pressure after 1 wk of exposure to hypoxia (20.1 +/- 1.8 mmHg) was significantly (P < 0.01, ANOVA) elevated compared with control conditions (12.1 +/- 0.1 mmHg) but was not altered in hypoxia hypercapnia (13.5 +/- 0.9 mmHg) or hypercapnia (11.8 +/- 1.3 mmHg). HPV (FI(O(2)) = 0.03) was attenuated in hypoxia, hypoxia-hypercapnia, and hypercapnia compared with control (P < 0.05, ANOVA). Addition of N(omega)-nitro-L-arginine methyl ester (10(-4) M), which augmented HPV in control, hypoxia, and hypercapnia, significantly reduced HPV in hypoxia-hypercapnia. Chronic hypoxia caused impaired endothelium-dependent relaxation in isolated pulmonary arteries, but coexistent hypercapnia partially protected against this effect. These findings suggest that coexistent hypercapnia inhibits hypoxia-induced pulmonary vascular remodeling and right ventricular hypertrophy, reduces HPV, and protects against hypoxia-induced impairment of endothelial function. PMID- 10666062 TI - Role of adenosine A(2B) receptors in vasodilation of rat pial artery and cerebral blood flow autoregulation. AB - This study was aimed to investigate the underlying mechanism of vasodilation induced by the activation of A(2B) adenosine receptors in relation to cerebral blood flow (CBF) autoregulation. Changes in pial arterial diameters were observed directly through a closed cranial window. N(omega)-nitro-L-arginine methyl ester (L-NAME, nitric oxide synthase inhibitor) significantly suppressed the concentration-dependent vasodilations induced by adenosine and 5'-N ethylcarboxamido-adenosine (NECA) but not the vasodilation by CGS-21680 (A(2A) receptor agonist). Moreover, NECA-induced vasodilation was suppressed by alloxazine (1 micromol/l) but not by ZM-241385 (1 micromol/l, A(2A) antagonist), which suggests mediation by A(2B)- receptor activation. Otherwise, the level of nitrite/nitrate was concentration dependently increased in the artificial cerebrospinal fluid (CSF) when adenosine and NECA were suffused over the cortical surface. L-NAME and alloxazine, but not ZM-241385, largely inhibited their releases. The lower limit of CBF autoregulation was little affected following pretreatment with L-NAME or alloxazine. Thus it is suggested that adenosine induced vasodilation via activation of A(2B)-adenosine receptors of the rat pial artery is coupled to the production of nitric oxide, which contributes little to CBF autoregulation. PMID- 10666063 TI - Transient increases in diameter and [Ca(2+)](i) are not obligatory for myogenic constriction. AB - Studies were performed to determine the significance of temporal variation in vascular smooth muscle Ca(2+) signaling during acute arteriolar myogenic constriction and, in particular, the importance of the stretch-induced intracellular Ca(2+) concentration ([Ca(2+)](i)) transient in attaining a steady state mechanical response. Rat cremaster arterioles (diameter approximately 100 microm) were dissected from surrounding tissues, and vessel segments were pressurized in the absence of intraluminal flow. For [Ca(2+)](i) measurements, vessels were loaded with fura 2 and fluorescence emitted by excitation at 340 and 380 nm was measured using video-based image analysis. Ca(2+) and diameter responses were examined after increases in intravascular pressure were applied as an acute step increase or a ramp function. Additional studies examined the effect of longitudinal vessel stretch on [Ca(2+)](i) and arteriolar diameter. Step increase in intraluminal pressure (from 50 to 120 mmHg) caused biphasic change in [Ca(2+)](i) and diameter. [Ca(2+)](i) transiently increased to 114.0 +/- 2.0% of basal levels and subsequently declined to 106.7 +/- 4.4% at steady state. Diameter initially distended to 125.4 +/- 2.1% of basal levels before constricting to 71.1 +/- 1.2%. In contrast, when the same pressure increase was applied as a ramp function (over 5 min) transient vessel distension and transient increase in [Ca(2+)](i) were prevented, yet at steady state vessels constricted to 71.3 +/- 2.5%. Longitudinal stretch resulted in a large [Ca(2+)](i) transient (158 +/- 19% of basal) that returned to baseline despite maintenance of the stretch stimulus. The data demonstrate that the initial vessel distension (reflecting myocyte stretch) and associated global [Ca(2+)](i) transient are not obligatory for myogenic contraction. Thus, although arteriolar smooth muscle cells are responsive to acute stretch, the resulting changes in myogenic tone may be more closely related to other mechanical variables such as wall tension. PMID- 10666064 TI - Functional role of angiotensin II type 1 and 2 receptors in regulation of uterine blood flow in nonpregnant sheep. AB - The objective was to determine the receptor subtype of angiotensin II (ANG II) that is responsible for vasoconstriction in the nonpregnant ovine uterine and systemic vasculatures. Seven nonpregnant estrogenized ewes with indwelling uterine artery catheters and flow probes received bolus injections (0.1, 0.3 and 1 microg) of ANG II locally into the uterine artery followed by a systemic infusion of ANG II at 100 ng x kg(-1) x min(-1) for 10 min to determine uterine vasoconstrictor responses. Uterine ANG II dose-response curves were repeated following administration of the ANG II type 2 receptor (AT(2)) antagonist PD 123319 and then repeated again in the presence of an ANG II type 1 receptor (AT(1)) antagonist L-158809. In a second experiment, designed to investigate the mechanism of ANG II potentiation that occurred in the presence of AT(2) blockade, nonestrogenized sheep received a uterine artery infusion of L-158809 (3 mg/min for 5 min) prior to the infusion of 0.03 microg/min of ANG II for 10 min. ANG II produced dose-dependent decreases in uterine blood flow (P < 0.03), which were potentiated in the presence of the AT(2) antagonist (P < 0.02). Addition of the AT(1) antagonist abolished the uterine vascular responses and blocked ANG II induced increases in systemic arterial pressure (P < 0.01). Significant uterine vasodilation (P < 0.01) was noted with AT(1) blockade in the second experiment, which was reversed by administration of the AT(2) antagonist or by the nitric oxide synthetase inhibitor N(omega)-nitro-L-arginine methyl ester. We conclude that the AT(1)-receptors mediate the systemic and uterine vasoconstrictor responses to ANG II in the nonpregnant ewe. AT(2)-receptor blockade resulted in a potentiation of the uterine vasoconstrictor response to ANG II, suggesting that the AT(2)-receptor subtype may modulate uterine vascular responses to ANG II potentially by release of nitric oxide. PMID- 10666065 TI - Regulation of myocardial blood flow response to mental stress in healthy individuals. AB - Mental stress testing has been proposed as a noninvasive tool to evaluate endothelium-dependent coronary vasomotion. In patients with coronary artery disease, mental stress can induce myocardial ischemia. However, even the determinants of the physiological myocardial blood flow (MBF) response to mental stress are poorly understood. Twenty-four individuals (12 males/12 females, mean age 49 +/- 13 yr, range 31-74 yr) with a low likelihood for coronary artery disease were studied. Serum catecholamines, cardiac work, and MBF (measured quantitatively with N-13 ammonia and positron emission tomography) were assessed. During mental stress (arithmetic calculation) MBF increased significantly from 0.70 +/- 0.14 to 0.92 +/- 0.21 ml x min(-1) x g(-1) (P < 0.01). Mental stress caused significant increases (P < 0.01) in serum epinephrine (26 +/- 16 vs. 42 +/ 17 pg/ml), norepinephrine (272 +/- 139 vs. 322 +/- 136 pg/ml), and cardiac work [rate-pressure product (RPP) 8,011 +/- 1,884 vs. 10,416 +/- 2,711]. Stress induced changes in cardiac work were correlated with changes in MBF (r = 0.72; P < 0.01). Multiple-regression analysis revealed stress-induced changes in the RPP as the only significant (P = 0.0001) predictor for the magnitude of mental stress induced increases in MBF in healthy individuals. Data from this group of healthy individuals should prove useful to investigate coronary vasomotion in individuals at risk for or with documented coronary artery disease. PMID- 10666066 TI - B(1) and B(2) bradykinin receptors on adventitial fibroblasts of cerebral arteries are coupled to recombinant eNOS. AB - Our previous ex vivo and in vivo studies reported that expression of the recombinant endothelial nitric oxide (NO) synthase (eNOS) gene in adventitial fibroblasts recovers NO production in arteries without endothelium in response to bradykinin. The present study was designed to characterize subtypes of bradykinin receptors on adventitial fibroblasts coupled to the activation of recombinant eNOS. Endothelium-denuded segments of canine basilar arteries were transduced with beta-galactosidase (beta-Gal) gene or eNOS gene ex vivo, using a replication defective adenoviral vector (10(10) plaque-forming units/ml) for 30 min at 37 degrees C. Twenty-four hours later, isometric force recording or cGMP measurement was carried out. B(1) bradykinin receptor agonist (des-Arg(9)-bradykinin, 10(-10) 10(-8) mol/l) did not significantly affect vascular tone in control or beta-Gal gene-transduced canine basilar arteries without endothelium. In contrast, this agonist caused concentration-dependent relaxations in recombinant eNOS gene transduced arteries without endothelium. Relaxations to B(1) receptor agonist in the eNOS arteries were abolished by B(1) receptor antagonist (des-Arg(9) [Leu(8)]bradykinin, 6 x 10(-9) mol/l) but not by B(2) receptor antagonist (Hoe 140, 5 x 10(-8) mol/l). Bradykinin did not significantly alter vascular tone in control or beta-gal arteries without endothelium, whereas this peptide (10(-11) 10(-8) mol/l) induced concentration-dependent relaxations, as well as an increase in cGMP formation in endothelium-denuded eNOS-transduced arteries. Stimulatory effects of bradykinin were prevented in the presence of a B(2) receptor antagonist but not in the presence of a B(1) receptor antagonist. B(1) and B(2) receptor antagonists had no effect on relaxations to substance P, confirming the selectivity of the compounds. Our results suggest that B(1) and B(2) bradykinin receptors are coupled to activation of recombinant eNOS expressed in adventitial fibroblasts. PMID- 10666068 TI - Dynamics of flow, resistance, and intramural vascular volume in canine coronary circulation. AB - Varying coronary volume will vary vascular resistance and thereby have an effect on coronary hemodynamics. Six ventricular septa were isolated from anesthetized dogs, dispersed in a biaxial stretch apparatus at diastolic stress, and perfused artificially with an oxygenated perfluorochemical emulsion at maximal vasodilation. Flow and thickness were measured continuously by an electromagnetic flow probe and sonomicrometer. Pressure was varied sinusoidally around 30, 50, and 70 mmHg with an amplitude of 7.5 mmHg; frequencies ranged between 0.015 and 7 Hz. Bode plots of admittance (flow/pressure) and capacitance (scaled thickness/pressure) were constructed. A two-compartment model was used in which the resistances vary with volume. Realistic values of microvascular compliance ( approximately 0.3 ml x mmHg(-1) x 100 g(-1)) were found. Values 10 times higher were then found when resistances were forced to be constant. We concluded that volume dependence of resistances have to be taken into account when dynamic or static pressure-flow relations are studied and conceal the effect of a large intramyocardial compliance on arterial hemodynamics. PMID- 10666067 TI - Tyrosine phosphorylation modulates arteriolar tone but is not fundamental to myogenic response. AB - The present study investigated the role of protein tyrosine phosphorylation in myogenic responsiveness of rat skeletal muscle arterioles. Arteriolar segments were cannulated and pressurized without intraluminal flow. All vessels studied developed spontaneous tone and demonstrated significant myogenic constriction to step changes in pressure with a resultant increase in myogenic tone over an intraluminal pressure range of 50-150 mmHg. Step increases in intraluminal pressure from 50 to 120 mmHg caused a rapid and sustained elevation in intracellular [Ca(2+)], as measured using fura 2. Vessels with myogenic tone dilated in response to tyrosine kinase inhibitors genistein (10 or 30 microM) and tyrphostin A47 (10 or 30 microM) and constricted to the tyrosine phosphatase inhibitor pervanadate (1 or 10 microM). Despite the dilator effect, myogenic reactivity was not blocked by the inhibitors. Daidzein (10 microM), a compound structurally similar to genistein but without tyrosine kinase-inhibiting activity, did not alter vessel tone or myogenic responses. Preincubation of arterioles with genistein or tyrphostin A47 did not significantly alter baseline arteriolar [Ca(2+)], and neither drug reduced the increase in [Ca(2+)] following an acute increase in intraluminal pressure. Constriction induced by pervanadate (10 microM) was not accompanied by a significant increase in intracellular [Ca(2+)], even though removal of extracellular Ca(2+) reversed the constriction. Examination of smooth muscle tyrosine phosphorylation, using a fluorescent phosphotyrosine antibody and confocal microscopy, showed that increased intraluminal pressure resulted in an increase in anti-phosphotyrosine fluorescence. Because manipulation of tyrosine kinase activity was found to alter vessel diameter, these data support a role for tyrosine phosphorylation in modulation of arteriolar tone. However, the results indicate that acute arteriolar myogenic constriction does not require tyrosine phosphorylation. PMID- 10666069 TI - VSM growth is stimulated in sympathetic neuron/VSM cocultures: role of TGF-beta2 and endothelin. AB - Sympathetic nerves are purported to stimulate blood vessel growth. The mechanism(s) underlying this stimulation has not been determined. With use of an in vitro coculture model, the present study tests the hypothesis that sympathetic neurons stimulate the growth of vascular smooth muscle (VSM) and evaluates potential mechanisms mediating this stimulation. Sympathetic neurons isolated from superior cervical ganglia (SCG) stimulated the growth of VSM. Growth of VSM in the presence of SCG (856 +/- 81%) was significantly greater than that in the absence of SCG (626 +/- 66%, P < 0.05). SCG did not stimulate VSM growth in transwell cocultures. An antibody that neutralized the activity of transforming growth factor-beta2 (TGF-beta2) inhibited SCG stimulation of VSM growth in coculture. SCG stimulation of VSM growth was also inhibited by an endothelin A receptor antagonist. These data suggest novel mechanisms for sympathetic modulation of vascular growth that may play a role in the physiological and/or pathological growth of the vasculature. PMID- 10666070 TI - Expression of the beta (slow)-isoform of MHC in the adult mouse heart causes dominant-negative functional effects. AB - Alpha- and beta-myosin heavy chain (MHC), the two MHC isoforms expressed in the mammalian heart, differ quantitatively in their enzymatic activities. The MHC composition of the heart can change dramatically in response to numerous stimuli, leading to the hypothesis that changes in cardiac function can be caused by myosin isoform shifts. However, this hypothesis has remained unproven because the stimuli used to generate these shifts are complex and accompanied by many additional physiological changes, including alterations in cardiac mass and geometry. Adult mouse ventricles normally express only alpha-MHC (the faster motor). To determine whether genetic alteration of the MHC isoform composition in the adult mouse heart would result in changes in cardiac chamber mass and contractility, we established transgenic mouse lines that express a Myc-tagged beta-MHC molecule (the slower motor) in adult ventricular tissue, one of which expresses 12% of its myosin as the transgene. There is no evidence of hypertrophy, induction of hypertrophic markers, and no histopathology. Myofibrillar Ca(2+)-activated ATPase activity is decreased by 23%, and Langendorff preparations demonstrate a significant 15% decrease in systolic function in transgenic hearts. These results suggest that even small shifts in the myosin isoform composition of the myocardium can result in physiologically significant changes in cardiac contractility and could be relevant to cardiovascular disease. PMID- 10666071 TI - Effects of recombinant eNOS gene expression on reactivity of small cerebral arteries. AB - Resistance arteries are an important target for vascular gene therapy because they play a key role in the regulation of tissue blood flow. The present study was designed to determine the effects of recombinant endothelial (e) nitric oxide synthase (NOS) gene expression on vasomotor reactivity of small brain stem arteries (internal diameter, 253 +/- 2.5 microm). Arterial rings were exposed ex vivo to an adenoviral vector (10(9) and 10(10) plaque-forming units/ml) encoding eNOS gene or beta-galactosidase gene. Twenty-four hours after transduction, vascular function was examined by isometric force studies. Transgene expression was evident mainly in adventitia. In arteries with endothelium transduced with eNOS gene but not with control beta-galactosidase gene, relaxations to bradykinin and substance P were significantly augmented. Removal of endothelium abolished relaxations to bradykinin and substance P in control and beta-galactosidase arteries. However, in endothelium-denuded arteries transduced with recombinant eNOS, bradykinin and substance P caused relaxations that were abolished in the presence of the NOS inhibitor N(G)-nitro-L-arginine methyl ester. In control arteries, endothelium removal augmented relaxations to the nitric oxide donors sodium nitroprusside and diethylamine NONOate. This augmentation was absent in eNOS gene-transduced arteries without endothelium. Our results suggest that, in small brain stem arteries, expression of recombinant eNOS increases biosynthesis of nitric oxide. Adventitia of small arteries is a good target for expression of recombinant eNOS. Genetically engineered adventitial cells may serve as a substitute source of nitric oxide in cerebral arteries with dysfunctional endothelium. PMID- 10666072 TI - Increased aortic stiffness assessed by pulse wave velocity in apolipoprotein E deficient mice. AB - Atherosclerosis develops and progresses spontaneously in apolipoprotein E knockout (apoE-KO) mice. A direct consequence of atherosclerosis is an increase in vascular stiffness. Pulse wave velocity (PWV) has been used to assess the stiffness of large vessels and was found to be increased in patients with atherosclerosis. In the present study, aortic stiffness was assessed by PWV in 4- and 13-mo-old apoE-KO mice and age-matched controls (C57BL/6J). In 13-mo-old apoE KO mice with extensive atherosclerotic lesions in the aorta (61 +/- 4%), PWV increased significantly (3.8 +/- 0.2 m/s) compared with controls (2.9 +/- 0.2 m/s). Endothelial nitric oxide (EDNO)-mediated vasorelaxation in response to ACh was markedly diminished in the aortic rings isolated from 13-mo-old apoE-KO mice compared with age-matched controls. In contrast, in 4-mo-old apoE-KO mice with only moderate atherosclerotic lesions in the aorta (23 +/- 5%), there were no significant changes in PWV and EDNO-mediated relaxation compared with controls. Blood pressure was not different among the four groups of mice. There were no significant differences in endothelium-independent vascular responses to sodium nitroprusside among different groups investigated. Histological evaluation revealed focal fragmentation of the elastic laminae in the aortic walls of 13-mo old apoE-KO mice. These results demonstrate for the first time that aortic stiffness determined by PWV increases in 13-mo-old apoE-KO mice. Endothelial dysfunction and elastic destruction in vascular wall caused by atherosclerosis may have contributed. PMID- 10666073 TI - Role of leukocytes and tissue-derived oxidants in short-term skeletal muscle ischemia-reperfusion injury. AB - The relative contribution of xanthine oxidase (XO) and leukocytes to tissue injury after short-term ischemia is unknown. In this study, we subjected three groups of rat spinotrapezius muscles to 30-min ischemia and 1-h reperfusion: 1) ischemia-reperfusion (I/R) + 0.9% saline, 2) I/R + superoxide dismutase, and 3) I/R + oxypurinol. A fourth group served as nonischemic control. We quantified the increase in resistance (%DeltaR) caused by leukocyte-capillary plugging concurrently with myocyte uptake of propidium iodide (PI) [expressed as no. of PI spots per total volume of perfused tissue (N(PI)/V)] and performed assays to quantify XO activity, thiobarbituric acid-reactive substances (TBARS), and myeloperoxidase (MPO). Groups 2 and 3 exhibited significant decreases in N(PI)/V relative to group 1. MPO levels and TBARS were similar among all groups, and mean %DeltaR was significantly reduced in groups 2 and 3 relative to group 1. However, elevated XO was observed in groups 1 and 2 relative to group 3 and nonischemic controls. These data are consistent with the hypothesis that XO, rather than toxic species produced by plugging or venule-adherent leukocytes, is responsible for postischemic damage in this model. PMID- 10666074 TI - Measurements of calcium transients in ventricular cells during discontinuous action potential conduction. AB - The L-type calcium current (I(Ca)) is important in sustaining propagation during discontinuous conduction. In addition, I(Ca) is altered during discontinuous conduction, which may result in changes in the intracellular calcium transient. To study this, we have combined the ability to monitor intracellular calcium concentration ([Ca(2+)](i)) in an isolated cardiac cell using confocal scanning laser fluorescence microscopy with our "coupling clamp" technique, which allows action potential propagation from the real cell to a real-time simulation of a model cell. Coupling a real cell to a model cell with a value of coupling conductance (G(C) = 8 nS) just above the critical value for action potential propagation results in both an increased amplitude and an increased rate of rise of the calcium transient. Similar but smaller changes in the calcium transient are caused by increasing G(C) to 20 nS. The increase of [Ca(2+)](i) by discontinuous conduction is less than the increase of I(Ca), which may indicate that much of [Ca(2+)](i) is the result of calcium released from the sarcoplasmic reticulum rather than the integration of I(Ca). PMID- 10666075 TI - Electrical interactions between a real ventricular cell and an anisotropic two dimensional sheet of model cells. AB - We have extended our "coupling clamp" technique, in which we couple a real cell to a real-time simulation of a model cell, to now incorporate a real cardiac cell as the central element of a two-dimensional sheet of model cells, in which the coupling conductances may be different in the x and y directions and a specific region of lack of coupling conductance may serve as a resistive barrier. We stimulated the real cell in the central location and determined the critical size of the real cell for successful activation of the entire sheet. We found that this critical size was decreased when anisotropy was present compared with the isotropic case and was further decreased when the central site of stimulation was close to the resistive barrier. The heart normally has some degree of anisotropy, and it has been shown that the remodeling that occurs in peri-infarction zones produces a particular loss of lateral connections compared with end-to-end connections among heart cells. We propose that the normal existence of anisotropy and enhancement of the degree of anisotropy both by loss of lateral gap junctions and the development of resistive barriers may play a facilitating role in the development of ectopic foci that may lead to cardiac arrhythmias. PMID- 10666076 TI - Reduced NO-dependent arteriolar dilation during the development of cardiomyopathy. AB - Our previous studies have suggested that there is reduced nitric oxide (NO) production in canine coronary blood vessels after the development of pacing induced heart failure. The goal of these studies was to determine whether flow induced NO-mediated dilation is altered in coronary arterioles during the development of heart failure. Subepicardial coronary arterioles (basal diameter 80 microm) were isolated from normal canine hearts, from hearts with dysfunction but no heart failure, and from hearts with severe cardiac decompensation. Arterioles were perfused at increasing flow or administered agonists with no flow in vitro. In arterioles from normal hearts, flow increased arteriolar diameter, with one-half of the response being NO dependent and one-half prostaglandin dependent. Shear stress-induced dilation was eliminated by removing the endothelium. Arterioles from normal hearts and hearts with dysfunction but no failure responded to increasing shear stress with dilation that reached a maximum at a shear stress of 20 dyn/cm(2). In contrast, arterioles from failing hearts showed a reduced dilation, reaching only 55% of the dilation seen in vessels of normal hearts at a shear stress of 100 dyn/cm(2). This remaining dilation was eliminated by indomethacin, suggesting that the NO-dependent component was absent in coronary microvessels after the development of heart failure. Similarly, agonist-induced NO-dependent coronary arteriolar dilation was markedly attenuated after the development of heart failure. After the development of severe dilated cardiomyopathy and heart failure, the NO-dependent component of both shear stress and agonist-induced arteriolar dilation is reduced or entirely absent. PMID- 10666077 TI - Spatial variations in endothelial barrier function in disturbed flows in vitro. AB - Hindered barrier function has been implicated in the initiation and progression of atherosclerosis, a disease of focal nature associated with altered hemodynamics. In this study, endothelial permeability to macromolecules and endothelial electrical resistance were investigated in vitro in monolayers exposed to disturbed flow fields that model spatial variations in fluid shear stress found at arterial bifurcations. After 5 h of flow, areas of high shear stress gradients showed a 5.5-fold increase in transendothelial transport of dextran (molecular weight 70,000) compared with no-flow controls. Areas of undisturbed fully developed flow, within the same monolayer, showed a 2.9-fold increase. Monolayer electrical resistance decreased with exposure to flow. The resistance measured during flow and the rate of change in monolayer resistance after removal of flow were lowest in the vicinity of flow reattachment (highest shear stress gradients). These results demonstrate that endothelial barrier function and permeability to macromolecules are regulated by spatial variations in shear stress forces in vitro. PMID- 10666078 TI - Relationship between nociceptin/orphanin FQ and cerebral hemodynamics after hypoxia-ischemia in piglets. AB - This study was designed to characterize the role of the newly described endogenous opioid nociceptin/orphanin FQ (NOC/oFQ) in reduced cerebral blood flow (CBF) observed after ischemia-reperfusion (I/R) and combined hypoxia and ischemia reperfusion (H-I/R), as a function of time after onset of reperfusion in newborn pigs equipped with a closed cranial window. Global cerebral ischemia (20 min) was induced via elevation of intracranial pressure, whereas hypoxia (10 min) decreased PO(2) to 35 +/- 3 mmHg with unchanged PCO(2). I/R elevated cerebrospinal fluid (CSF) NOC/oFQ from 67 +/- 4 to 266 +/- 29 pg/ml within 1 h, whereas values returned to control level within 4 h of reperfusion. H-I/R elevated CSF NOC/oFQ to 483 +/- 67 pg/ml within 1 h, and such values returned slowly to control level within 12 h of reperfusion. Topical NOC/oFQ (10(-8) M, 10(-6) M)-induced vasodilation was attenuated by I/R and reversed to vasoconstriction by H-I/R at 1 h of reperfusion (control, 9 +/- 1 and 16 +/- 1%; I/R, 3 +/- 1 and 6 +/- 1%; H-I/R, -6 +/- 1 and -11 +/- 1%). Such altered dilation returned to control values within 4 h in I/R animals and within 12 h in H-I/R animals. Blood flow in the cerebrum was reduced from 58 +/- 4 to 33 +/- 2 ml x min(-1) x 100 g(-1) within 1 h and returned to control value within 4 h in I/R animals. In animals pretreated with [F/G]NOC/oFQ(1-13)-NH(2) (1 mg/kg iv), an NOC/oFQ antagonist, however, CBF only fell to 43 +/- 3 ml x min(-1) x 100 g(-1) at 1 h of reperfusion. Similar observations were made in H-I/R animals. These data suggest that an elevated CSF NOC/oFQ concentration and altered vascular responsiveness to this opioid contribute to reductions in CBF observed after either I/R or H-I/R. PMID- 10666079 TI - Developmental differences in delayed rectifying outward current in feline ventricular myocytes. AB - In the present work, we found that the delayed rectifying outward potassium current (I(K)) in adult and neonatal cat ventricular myocytes consists of both rapid and slow components, I(Kr) and I(Ks), respectively, which can be isolated pharmacologically. Thus after complete blockade of I(Kr) with dofetilide, the remaining I(Ks) current is homogeneous, as shown by an envelope of tails test. I(Kr) maximum tail current density, measured at -40 mV, was similar in adult and neonatal myocytes. I(Ks) maximum tail current density in neonatal myocytes, measured at -40 mV, was significantly smaller than in adult myocytes. Activation kinetics of I(Kr) and I(Ks) was similar in both age groups. However, the I(Kr) deactivation time course was significantly faster in neonatal than in adult myocytes. Developmental differences in the subunit composition of I(Kr) that display distinctly different deactivation kinetics are suggested. PMID- 10666080 TI - Role of endothelin receptor subtypes in volume-stimulated ANF secretion. AB - The role of endothelin (ET) receptors was tested in volume-stimulated atrial natriuretic factor (ANF) secretion in conscious rats. Mean ANF responses to slow infusions (3 x 3.3 ml/8 min) were dose dependently reduced (P < 0.05) by bosentan (nonselective ET-receptor antagonist) from 64.1 +/- 18.1 (SE) pg/ml (control) to 52.6 +/- 16.1 (0.033 mg bosentan/rat), 16.1 +/- 7.6 (0. 33 mg/rat), and 11.6 +/- 6.5 pg/ml (3.3 mg/rat). The ET-A-receptor antagonist BQ-123 (1 mg/rat) had no effect relative to DMSO controls, whereas the putative ET-B antagonist IRL-1038 (0.1 mg/rat) abolished the response. In a second protocol, BQ-123 (>/=0.5 mg/rat) nonsignificantly reduced the peak ANF response (106.1 +/- 23.0 pg/ml) to 74.0 +/- 20.5 pg/ml for slow infusions (3.5 ml/8.5 min) but reduced the peak response (425.3 +/- 58.1 pg/ml) for fast infusions (6.6 ml/1 min) by 49.9% (P < 0.001) and for 340 pmoles ET-1 (328.8 +/- 69.5 pg/ml) by 83.5% (P < 0.0001). BQ-123 abolished the ET-1-induced increase in arterial pressure (21.8 +/- 5.2 mmHg at 1 min). Changes in central venous pressure were similar for DMSO and BQ-123 (slow: 0.91 and 1.14 mmHg; fast: 4.50 and 4.13 mmHg). The results suggest 1) ET-B receptors mainly mediate the ANF secretion to slow volume expansions of <1.6%/min; and 2) ET-A receptors mainly mediate the ANF response to acute volume overloads. PMID- 10666081 TI - Elevated salt intake impairs dilation of rat skeletal muscle resistance arteries via ANG II suppression. AB - Vasodilator responses were assessed in resistance arteries (100-200 microm) isolated from the gracilis muscle of normotensive rats after changes in dietary salt intake. Sprague-Dawley rats were maintained on either a high-salt (HS) diet (4.0% NaCl) or a low-salt (LS) diet (0.4% NaCl) for 4-8 wk (chronic) or 3 days (short-term) with water ad libitum. One group of short-term HS rats received a continuous intravenous infusion of a low dose (5 ng x kg(-1) x min(-1)) of ANG II to prevent the ANG II suppression that occurs with HS diet. Short-term and chronic HS diet eliminated arterial dilation in response to ACh and reduced PO(2) (30-40 mmHg) and the stable prostacyclin analog iloprost. ANG II infusion preserved the response to these vasodilator stimuli in short-term HS animals. Dilator responses to sodium nitroprusside and forskolin were unaffected by HS diet. These findings suggest that ANG II suppression during HS diet impairs vascular relaxation mechanisms upstream from the cAMP and cGMP second messenger systems. PMID- 10666082 TI - Role of kinins in chronic heart failure and in the therapeutic effect of ACE inhibitors in kininogen-deficient rats. AB - Using Brown Norway Katholiek (BNK) rats, which are deficient in kininogen (kinin precursor) due to a mutation in the kininogen gene, we examined the role of endogenous kinins in 1) normal cardiac function; 2) myocardial infarction (MI) caused by coronary artery ligation; 3) cardiac remodeling in the development of heart failure (HF) after MI; and 4) the cardioprotective effect of angiotensin converting enzyme inhibitors (ACEI) on HF after MI. Two months after MI, rats were randomly treated with vehicle or the ACEI ramipril for 2 mo. Brown Norway rats (BN), which have normal kininogen, were used as controls. Left ventricular (LV) end-diastolic volume (EDV), end-systolic volume (ESV), end-diastolic pressure (EDP), and ejection fraction (EF) as well as myocardial infarct size (IS), interstitial collagen fraction (ICF), cardiomyocyte cross-sectional area (MCA), and oxygen diffusion distance (ODD) were measured. We found that 1) cardiac hemodynamics, function, and histology were the same in sham-ligated BN and BNK rats; 2) IS was similar in BN and BNK; 3) in rats with HF treated with vehicle, the decrease in LVEF and the increase in LVEDV, LVESV, LVEDP, ICF, MCA, and ODD did not differ between BNK and BN; and 4) ACEI increased EF, decreased LVEDV and LVESV, and improved cardiac remodeling in BN-HF rats, and these effects were partially blocked by the bradykinin B(2) receptor antagonist icatibant (HOE 140). In BNK-HF rats, ACEI failed to produce these beneficial cardiac effects. We concluded that in rats, lack of kinins does not influence regulation of normal cardiac function, myocardial infarct size, or development of HF; however, kinins appear to play an important role in the cardioprotective effect of ACEI, since 1) this effect was significantly diminished in kininogen-deficient rats and 2) it was blocked by a B(2) kinin receptor antagonist in BN rats. PMID- 10666083 TI - Effects of autonomic disruption and inactivity on venous vascular function. AB - The effects of autonomic disruption and inactivity were studied on the venous vascular system. Forty-eight subjects, 24 with spinal cord injury (SCI) and 12 sedentary and 12 active able-bodied controls, participated in this study. Peripheral autonomic data were obtained to estimate sympathetic vasomotor control [low-frequency component of systolic blood pressure (LF(SBP))]. Vascular parameters were determined using strain-gauge venous occlusion plethysmography: venous capacitance (VC), venous emptying rate (VER), and total venous outflow (VO(t)). An additional vascular parameter was calculated: venous compliance [(VC/occlusion pressure) x 100]. VC and VO(t) were significantly different (SCI < sedentary < active). VER adjusted for VC was not different for any group comparison, whereas venous compliance was significantly lower in the SCI group than in the able-bodied groups and in the sedentary group compared with the active group. Regression analysis for the total group revealed a significant relationship between LF(SBP) and venous compliance (r = 0.64, P < 0.0001). After controlling for LF(SBP) through analysis of covariance, we found that mean differences for all venous vascular parameters did not change from unadjusted mean values. Our findings suggest that in subjects with SCI, the loss of sympathetic vasomotor tone contributes more than inactivity to reductions in venous vascular function. Heightened VC, VO(t), vasomotor tone, and venous compliance in the active group compared with the sedentary group imply that regular endurance training contributes to optimal venous vascular function and peripheral autonomic integrity. PMID- 10666084 TI - Mechanical stretch stimulates growth of vascular smooth muscle cells via epidermal growth factor receptor. AB - We have studied whether activation of epidermal growth factor receptor (EGFR) is involved in stretch-induced extracellular signal-regulated kinase 1/2 (ERK1/2) activation and protein synthesis in cultured rat vascular smooth muscle cells (VSMC). Cyclic stretch (1 Hz) induced a rapid (within 5 min) phosphorylation of ERK1/2, an effect that was time and strength dependent and inhibited by an EGFR kinase inhibitor (AG-1478) but not by a platelet-derived growth factor receptor kinase inhibitor (AG-1296). The stretch rapidly (within 2 min) induced tyrosine phosphorylation of several proteins, among which 180-kDa protein was shown to be EGFR as revealed by blockade with AG-1478 as well as immunoprecipitation with anti-EGFR antibody coupled with immunoblotting with anti-phosphotyrosine antibody. The stretch rapidly (within 2 min) induced association of tyrosine phosphorylated EGFR with adaptor proteins (Shc/Grb2) as revealed by coprecipitation with glutathione-S-transferase-Grb2 fusion protein coupled with immunoblotting with anti-phosphotyrosine, anti-EGFR, and anti-Shc antibodies. Transfection of a dominant-negative mutant of H-Ras also inhibited stretch induced ERK1/2 activation. Treatment with a stretch-activated ion channel blocker (Gd(3+)) and an intracellular Ca(2+) antagonist (TMB-8) inhibited stretch-induced phosphorylation of EGFR and ERK1/2. Treatment with AG-1478 and a mitogen activated protein kinase kinase inhibitor (PD-98059), but not AG-1296, blocked [(3)H]leucine uptake stimulated by a high level of stretch. These data suggest that ERK1/2 activation by mechanical stretch requires Ca(2+)-sensitive EGFR activation mainly via stretch-activated ion channels, thereby leading to VSMC growth. PMID- 10666085 TI - Muscle metaboreflex control of cardiac output and peripheral vasoconstriction exhibit different latencies. AB - Experiments were designed to determine 1) the mechanisms mediating metaboreflex induced increases in systemic arterial pressure (SAP) in response to total vascular occlusion of hindlimb blood flow [e.g., increases in cardiac output (CO) vs. peripheral vasoconstriction] and 2) whether the individual mechanisms display differential latencies for the onset of the responses. Responses were observed in seven dogs performing steady-state treadmill exercise of mild and moderate workloads (3.2 km/h at 0% grade and 6.4 km/h at 10% grade). Differential latencies were exhibited among CO, nonischemic vascular conductance (NIVC; conductance to all nonischemic vascular beds), and renal vascular conductance (RVC), with peripheral vasoconstriction significantly preceding metaboreflex mediated increases in CO. In addition, the latencies for SAP were not different from those for NIVC or RVC at either workload. During the lower workload there were small increases and then subsequent decreases in CO before the metaboreflex induced increase in CO, which did contribute somewhat to the initial increases in SAP. However, the increases in CO mediated by the metaboreflex occurred significantly later than the initial increases in SAP. Therefore, we conclude that the substantial metaboreflex-mediated pressor responses that occur during the initial phase of total vascular occlusion during mild and moderate exercise are primarily caused by peripheral vasoconstriction. PMID- 10666086 TI - Increased urinary excretion of uroguanylin in patients with congestive heart failure. AB - Uroguanylin is a small-molecular-weight peptide that activates membrane-bound receptor-guanylate cyclases in the intestine, kidney, and other epithelia. Uroguanylin has been shown to participate in the regulation of salt and water homeostasis in mammals via cGMP-mediated processes, bearing a distinct similarity to the action of the atriopeptins, which play a defined role in natriuresis and act as prognostic indicators of severe congestive heart failure (CHF). The objectives of this study were to measure the urinary levels of uroguanylin and the circulating plasma levels of atrial natriuretic peptide (ANP) in healthy individuals (n = 53) and patients with CHF (n = 16). Urinary excretion of uroguanylin was assessed by a cGMP accumulation bioassay employing human T84 intestinal cells. In individuals without CHF, the concentration of uroguanylin bioactivity was 1.31 +/- 0.27 nmol cGMP/ml urine and 1.73 +/- 0.25 micromol cGMP/24-h urine collection. The urinary bioactivity of uroguanylin in males (1.74 +/- 0.55 nmol cGMP/ml urine; n = 27) tended to be higher than the excretion levels in females (0.94 +/- 0.16 nmol cGMP/ml urine; n = 26) over a 24-h period but did not achieve statistical significance. Both male and female groups showed 24-h temporal diurnal variations with the highest uroguanylin levels observed between the hours of 8:00 AM and 2:00 PM. The circulating level of ANP was 12.1 +/- 1.6 pg/ml plasma and did not significantly vary with respect to male/female population or diurnal variation. In patients with CHF, the concentration of plasma ANP and urinary uroguanylin bioactivity increased substantially (7.5-fold and 70-fold, respectively, both P right coronary artery > left anterior descending coronary artery > aorta. The isolation of coronary microvessels from the left ventricle was associated with an enrichment of endothelial cell markers such as eNOS, von Willebrand factor, and caveolin-1, an observation supported by the detection of up to 15-fold higher levels of eNOS mRNA in coronary microvessels relative to the larger arteries. The ability to quantify changes in eNOS mRNA levels throughout the canine vasculature should provide greater insight into the molecular mechanisms of how this gene is regulated in physiological and pathophysiological states. PMID- 10666100 TI - Caffeine-induced Ca(2+) sparks in mouse ventricular myocytes. AB - Ca(2+) sparks are spatially localized intracellular Ca(2+) release events that were first described in 1993. Sparks have been ascribed to sarcoplasmic reticulum Ca(2+) release channel (ryanodine receptor, RyR) opening induced by Ca(2+) influx via L-type Ca(2+) channels or by spontaneous RyR openings and have been thought to reflect Ca(2+) release from a cluster of RyR. Here we describe a pharmacological approach to study sparks by exposing ventricular myocytes to caffeine with a rapid solution-switcher device. Sparks under these conditions have properties similar to naturally occurring sparks in terms of size and intracellular Ca(2+) concentration ([Ca(2+)](i)) amplitude. However, after the diffusion of caffeine, sparks first appear close to the cell surface membrane before coalescing to produce a whole cell transient. Our results support the idea that a whole cell [Ca(2+)](i) transient consists of the summation of sparks and that Ca(2+) sparks consist of the opening of a cluster of RyR and confirm that characteristics of the cluster rather than the L-type Ca(2+) channel-RyR relation determine spark properties. PMID- 10666101 TI - Tyrosine kinase and protein kinase C regulate L-type Ca(2+) current cooperatively in human atrial myocytes. AB - The effects of tyrosine protein kinases (TK) on the L-type Ca(2+) current (I(Ca)) were examined in whole cell patch-clamped human atrial myocytes. The TK inhibitors genistein (50 microM), lavendustin A (50 microM), and tyrphostin 23 (50 microM) stimulated I(Ca) by 132 +/- 18% (P < 0.001), 116 +/- 18% (P < 0.05), and 60 +/- 6% (P < 0.001), respectively. After I(Ca) stimulation by genistein, external application of isoproterenol (1 microM) caused an additional increase in I(Ca). Dialyzing the cells with a protein kinase A inhibitor suppressed the effect of isoproterenol on I(Ca) but not that of genistein. Inhibition of protein kinase C (PKC) by pretreatment of cells with 100 nM staurosporine or 100 nM calphostin C prevented the effects of genistein on I(Ca). The PKC activator phorbol 12-myristate 13-acetate (PMA), after an initial stimulation (75 +/- 17%, P < 0.05), decreased I(Ca) (-36 +/- 5%, P < 0.001). Once the inhibitory effect of PMA on I(Ca) had stabilized, genistein strongly stimulated the current (323 +/- 25%, P < 0.05). Pretreating myocytes with genistein reduced the inhibitory effect of PMA on I(Ca). We conclude that, in human atrial myocytes, TK inhibit I(Ca) via a mechanism that involves PKC. PMID- 10666103 TI - An appreciation: eugene rannels PMID- 10666102 TI - Seven years at the wheel PMID- 10666104 TI - How does cAMP increase active Na absorption across alveolar epithelium? PMID- 10666105 TI - Mechanisms of increased Na(+) transport in ATII cells by cAMP: we agree to disagree and do more experiments. AB - Existing evidence supports the presence of active transport of Na(+) across the mammalian alveolar epithelium and its upregulation by agents that increase cytoplasmic cAMP levels. However, there is controversy regarding the mechanisms responsible for this upregulation. Herein we present the results of various patch clamp studies indicating the presence of 25- to 27-pS, amiloride-sensitive, moderately selective Na(+) channels (Na(+)-to-K(+) permeability ratio = 7:1) located on the apical membranes of rat alveolar type II (ATII) cells maintained in primary culture. The addition of terbutaline to the bath solution increased the open probability of single channels present in cell-attached patches of ATII cells without affecting their conductance. A similar increase in open probability was seen after the addition of protein kinase A, ATP, and Mg(2+) to the cytoplasmic side of inside-out patches. Measurement of short-circuit currents across confluent monolayers of rat or rabbit ATII cells indicates that terbutaline and 8-(4-chlorophenylthio)-cAMP increase vectorial Na(+) transport and activate Cl(-) channels. Currently, there is a controversy as to whether the cAMP-induced increase in Na(+) transport is due solely to hyperpolarization of the cytoplasmic side of the ATII cell membrane due to Cl(-) influx or whether it results from simultaneous stimulation of both Cl(-) and Na(+) conductive pathways. Additional studies are needed to resolve this issue. PMID- 10666106 TI - Cl-channel activation is necessary for stimulation of Na transport in adult alveolar epithelial cells. AB - In this review, we discuss evidence that supports the hypothesis that adrenergic stimulation of transepithelial Na absorption across the alveolar epithelium occurs indirectly by activation of apical Cl channels, resulting in hyperpolarization and an increased driving force for Na uptake through amiloride sensitive Na channels. This hypothesis differs from the prevailing idea that adrenergic-receptor activation increases the open probability of Na channels, leading to an increase in apical membrane Na permeability and an increase in Na and fluid uptake from the alveolar space. We review results from cultured alveolar epithelial cell monolayer experiments that show increases in apical membrane Cl conductance in the absence of any change in Na conductance after stimulation by selective beta-adrenergic-receptor agonists. We also discuss possible reasons for differences in Na-channel regulation in cells grown in monolayer culture compared with that in dissociated alveolar epithelial cells. Finally, we describe some preliminary in vivo data that suggest a role for Cl channel activation in the process of amiloride-sensitive alveolar fluid absorption. PMID- 10666107 TI - Effect of steroid on hyperoxia-induced ICAM-1 expression in pulmonary endothelial cells. AB - Intercellular adhesion molecule-1 (ICAM-1) of the vascular endothelium plays a key role in the development of pulmonary oxygen toxicity. We studied the effect of steroid on hyperoxia-induced ICAM-1 expression using cultured endothelial cells in vitro. Human pulmonary artery endothelial cells (HPAECs) were cultured to confluence, and then the monolayers were exposed to either control (21% O(2) 5% CO(2)) or hyperoxic (90% O(2)-5% CO(2)) conditions with and without a synthetic glucocorticoid, methylprednisolone (MP). MP reduced hyperoxia-induced ICAM-1 and ICAM-1 mRNA expression in a dose-dependent manner. Neutrophil adhesion to hyperoxia-exposed endothelial cells was also inhibited by MP treatment. In addition, MP attenuated hyperoxia-induced H(2)O(2) production in HPAECs as assessed by flow cytometry. An electrophoretic mobility shift assay demonstrated that hyperoxia activated nuclear factor-kappaB (NF-kappaB) but not activator protein-1 (AP-1) and that MP attenuated hyperoxia-induced NF-kappaB activation dose dependently. With Western immunoblot analysis, IkappaB-alpha expression was decreased by hyperoxia and increased by MP treatment. These results suggest that MP downregulates hyperoxia-induced ICAM-1 expression by inhibiting NF-kappaB activation via increased IkappaB-alpha expression. PMID- 10666108 TI - Hyperoxia synergistically increases TNF-alpha-induced interleukin-8 gene expression in A549 cells. AB - Interleukin (IL)-8 is an important mediator of acute lung injury. Hyperoxia induces IL-8 production in some cell types, but its effect on IL-8 gene expression in respiratory epithelium is not well described. In addition, IL-8 gene expression resulting from the combined effects of hyperoxia and proinflammatory cytokines has not been well characterized. We treated cultured respiratory epithelial-like cells (A549 cells) with hyperoxia alone, tumor necrosis factor (TNF)-alpha alone, or the combination of TNF-alpha and hyperoxia and evaluated IL-8 gene expression. Hyperoxia alone had a minimal effect on IL-8 gene expression, and TNF-alpha alone increased IL-8 gene expression in a time dependent manner. In contrast, the combination of TNF-alpha and hyperoxia synergistically increased IL-8 gene expression as measured by ELISA (TNF-alpha alone for 24 h = 769 +/- 89 pg/ml vs. hyperoxia + TNF-alpha for 24 h = 1, 189 +/- 89 pg/ml) and Northern blot analyses. Experiments involving IL-8 promoter reporter assays, electromobility shift assays, and Western blot analyses demonstrated that hyperoxia augmented TNF-alpha-mediated activation of the IL-8 promoter by a nuclear factor (NF)-kappaB-dependent mechanism and increased the duration of NF-kappaB nuclear translocation after concomitant treatment with TNF alpha. Additional reporter gene assays demonstrated, however, that increased activation of NF-kappaB does not fully account for the synergistic effect of hyperoxia and that the NF-IL-6 site in the IL-8 promoter is also required for the synergistic effect of hyperoxia. We conclude that hyperoxia alone has a minimal effect on IL-8 gene expression but synergistically increases IL-8 gene expression in the presence of TNF-alpha by a mechanism involving cooperative interaction between the transcription factors NF-kappaB and NF-IL-6. PMID- 10666109 TI - High K(+)-induced membrane depolarization attenuates endothelium-dependent pulmonary vasodilation. AB - Impairment of endothelium-dependent pulmonary vasodilation has been implicated in the development of pulmonary hypertension. Pulmonary vascular smooth muscle cells and endothelial cells communicate electrically through gap junctions; thus, membrane depolarization in smooth muscle cells would depolarize endothelial cells. In this study, we examined the effect of prolonged membrane depolarization induced by high K(+) on the endothelium-dependent pulmonary vasodilation. Isometric tension was measured in isolated pulmonary arteries (PA) from Sprague Dawley rats, and membrane potential was measured in single PA smooth muscle cells. Increase in extracellular K(+) concentration from 4.7 to 25 mM significantly depolarized PA smooth muscle cells. The 25 mM K(+)-mediated depolarization was characterized by an initial transient depolarization (5-15 s) followed by a sustained depolarization that could last for up to 3 h. In endothelium-intact PA rings, ACh (2 microM), levcromakalim (10 microM), and nitroprusside (10 microM) reversibly inhibited the 25 mM K(+)-mediated contraction. Functional removal of endothelium abolished the ACh-mediated relaxation but had no effect on the levcromakalim- or the nitroprusside-mediated pulmonary vasodilation. Prolonged ( approximately 3 h) membrane depolarization by 25 mM K(+) significantly inhibited the ACh-mediated PA relaxation (-55 +/- 4 vs. 29 +/- 2%, P < 0.001), negligibly affected the levcromakalim-mediated pulmonary vasodilation (-92 +/- 4 vs. -95 +/- 5%), and slightly but significantly increased the nitroprusside-mediated PA relaxation (-80 +/- 2 vs. 90 +/- 3%, P < 0. 05). These data indicate that membrane depolarization by prolonged exposure to high K(+) concentration selectively inhibited endothelium-dependent pulmonary vasodilation, suggesting that membrane depolarization plays a role in the impairment of pulmonary endothelial function in pulmonary hypertension. PMID- 10666110 TI - PAF-induced synthesis of tetraenoic and pentaenoic leukotrienes in the isolated rabbit lung. AB - In an isolated rabbit lung model, we tested the hypothesis that platelet activating factor (PAF)-induced leukotriene (LT) synthesis is critically dependent on the free precursor fatty acid supply and the possible substitution of arachidonic acid (AA) by eicosapentaenoic acid (EPA). To augment the intravascular polymorphonuclear neutrophils (PMNs) in the isolated lung, human PMNs were infused into the pulmonary artery. LTs and hydroxyeicosatetra(penta)enoic acids were quantified with HPLC techniques. Application of PAF (5 microM) or AA (10 microM) provoked the generation of limited quantities of 4-series LTs and 5-hydroxyeicosatetraenoic acid (total sum of 5-lipoxygenase products approximately 7 and approximately 27 pmol/ml in lungs both with and without infused PMNs, respectively). Combined administration amplified 5-lipoxygenase product formation, with a predominance of cysteinyl-LT synthesis in lungs both without (total sum approximately 67 pmol/ml) and, much more strikingly, with (total sum approximately 308 pmol/ml) an infusion of neutrophils. EPA (10 microM) elicited exclusive generation of 5-series LTs and 5 hydroxyeicosapentaenoic acid (total sum approximately 82 pmol/ml). Dual stimulation with PAF and EPA provoked amplification of EPA-derived 5-lipoxygenase product formation, again with predominance of cysteinyl-LTs in lungs without (total sum approximately 224 pmol/ml) and, in particular, with (total sum approximately 545 pmol/ml) preceding microvascular PMN entrapment. Combined application of PAF, AA, and EPA resulted in the synthesis of LTs derived from both fatty acids, with a predominance of 5-series products. We conclude that the PAF-evoked 5-lipoxygenase product formation in the neutrophil-harboring lung capillary bed is critically dependent on intravascular precursor fatty acid supply, with EPA representing the preferred substrate compared with AA. PMN related transcellular eicosanoid synthesis is suggested to underlie the predominant generation of cysteinyl-LTs. The supply of n-3 versus n-6 precursor fatty acid may thus have a major impact on inflammatory mediator generation. PMID- 10666111 TI - Decreased synthesis and vasodilation to nitric oxide in piglets with hypoxia induced pulmonary hypertension. AB - Nitric oxide (NO) is thought to play an important role in the regulation of neonatal pulmonary vasculature. It has been suggested that neonates with pulmonary hypertension have a defective NO pathway. Therefore, we measured in 1 day-old piglets exposed to hypoxia (fraction of inspired O(2) = 0.10) for 3 or 14 days to induce pulmonary hypertension 1) the activity of NO synthase (NOS) via conversion of L-arginine to L-citrulline and the concentration of the NO precursor L-arginine in isolated pulmonary vessels, 2) the vasodilator response to the NO donor 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1) and the cGMP analog 8-bromo-cGMP in isolated perfused lungs, and 3) the production of cGMP in response to SIN-1 in isolated perfused lungs. After 3 days of exposure to hypoxia, endothelial NOS (eNOS) activity was unaffected, whereas, after 14 days of hypoxia, eNOS activity was decreased in the cytosolic fraction of pulmonary artery (P < 0.05) but not of pulmonary vein homogenates. Inducible NOS activity was decreased in the cytosolic fraction of pulmonary artery homogenates after both 3 (P < 0.05) and 14 (P < 0.05) days of hypoxia but was unchanged in pulmonary veins. Pulmonary artery levels of L-arginine were unaffected by hypoxic exposure. After 3 days of exposure to hypoxia, the reduction in the dilator response to SIN-1 (P < 0.05) coincided with a decrease in cGMP production (P < 0.005), suggesting that soluble guanylate cyclase activity may be altered. When the exposure was prolonged to 14 days, dilation to SIN-1 remained decreased (P < 0.05) and, although cGMP production normalized, the dilator response to 8-bromo cGMP decreased (P < 0.05), suggesting that, after prolonged exposure to hypoxia, cGMP-dependent mechanisms may also be impaired. In conclusion, neonatal hypoxia induced pulmonary hypertension is associated with multiple disruptions in the NO pathway. PMID- 10666112 TI - cDNA-RDA of genes expressed in fetal and adult lungs identifies factors important in development and function. AB - The identification of genetic factors important in lung development and function will help in understanding the underlying molecular mechanisms of respiratory disease. Representational difference analysis of cDNA (cDNA-RDA) is a PCR-based subtractive enrichment procedure for the isolation of differentially expressed genes. We performed cDNA-RDA and isolated genes expressed more abundantly in fetal and adult lungs. Fifty-four clones potentially representing genes with higher transcript levels in the fetal lung were sequenced. Sequence similarity searches indicated that these clones included 12 known genes, a discoidin-like domain-containing gene, six expressed sequence tags (ESTs), and one novel sequence. Fifty-six clones potentially representing genes expressed more abundantly in the adult lung were also cloned and sequenced. Of these, 16 known human genes were represented along with two sequences significantly similar to known mouse genes and two novel sequences. Several of these known genes are implicated in stress response and lung protection. Thus cDNA-RDA was successfully used to isolate known and novel differentially expressed genes, which putatively play an important role in human lung development. PMID- 10666113 TI - Energy state, pH, and vasomotor tone during hypoxia in precontracted pulmonary and femoral arteries. AB - To assess effects of smooth muscle energy state and intracellular pH (pH(i)) on pulmonary arterial tone during hypoxia, we measured ATP, phosphocreatine, P(i), and pH(i) by (31)P-NMR spectroscopy and isometric tension in phenylephrine contracted rings of porcine proximal intrapulmonary arteries. Hypoxia caused early transient contraction followed by relaxation and late sustained contraction. Energy state and pH(i) decreased during relaxation and recovered toward control values during late contraction. Femoral arterial rings had higher energy state and lower pH(i) under baseline conditions and did not exhibit late contraction or recovery of energy state and pH(i) during hypoxia. In pulmonary arteries, glucose-free conditions abolished late hypoxic contraction and recovery of energy state and pH(i), but endothelial denudation abolished only late hypoxic contraction. NaCN had little effect at 0. 1 and 1.0 mM but caused marked vasorelaxation and decreases in energy state and pH(i) at 10 mM. These results suggest that 1) regulation of tone, energy state, and pH(i) differed markedly in pulmonary and femoral arterial smooth muscle, 2) hypoxic relaxation was mediated by decreased energy state or pH(i) due to hypoxic inhibition of oxidative phosphorylation, 3) recovery of energy state and pH(i) in hypoxic pulmonary arteries was due to accelerated glycolysis mediated by mechanisms intrinsic to smooth muscle, and 4) late hypoxic contraction in pulmonary arteries was mediated by endothelial factors that required hypoxic recovery of energy state and pH(i) for transduction in smooth muscle or extracellular glucose for production and release by endothelium. PMID- 10666114 TI - Phospholipid molecular species of bronchoalveolar lavage fluid after local allergen challenge in asthma. AB - Electrospray ionization mass spectrometry was used to quantify phosphatidylcholine (PC) and phosphatidylglycerol (PG) molecular species in bronchoalveolar lavage fluid (BALF) from control and mild asthmatic subjects after local allergen challenge. BALF was obtained from 5 control and 13 asthmatic subjects before and 24 h after segmental allergen and saline challenge. There were no differences in the ratio of total PC to total PG or in the molecular species composition of PC or PG between the asthmatic and control groups under basal conditions. Allergen challenge in asthmatic but not in control volunteers caused a significant increase in the PC-to-PG ratio because of increased concentrations of PC species containing linoleic acid (16:0/18:2 PC, 18:0/18:2 PC, and 18:1/18:2 PC). These molecular species were characteristic of plasma PC analyzed from the same subjects, strongly suggesting that the altered PC composition in BALF in asthmatic subjects after allergen challenge was due to infiltration of plasma lipoprotein, not to catabolism of surfactant phospholipid. Interactions between surfactant and lipoprotein infiltrate may contribute to surfactant dysfunction and potentiate disease severity in asthma. PMID- 10666115 TI - Heme oxygenase-1 inhibits TNF-alpha-induced apoptosis in cultured fibroblasts. AB - Heme oxygenase (HO)-1 catalyzes the oxidative cleavage of heme to yield equimolar amounts of biliverdin, iron, and carbon monoxide. HO-1 is a stress response protein, the induction of which is associated with protection against oxidative stress. The mechanism(s) of protection is not completely elucidated, although it is suggested that one or more of the catalytic by-products provide antioxidant functions either directly or indirectly. The involvement of reactive oxygen species in apoptosis raised the question of a possible role for HO-1 in programmed cell death. Using the tetracycline-regulated expression system, we show here that conditional overexpression of HO-1 prevents tumor necrosis factor alpha-induced apoptosis in murine L929 fibroblasts. Inhibition of apoptosis was not observed in the presence of tin protoporphyrin, a specific inhibitor of HO activity, and in cells overexpressing antisense HO-1. Interestingly, exogenous administration of a low concentration of carbon monoxide also prevented tumor necrosis factor-alpha-induced apoptosis in L929 fibroblasts. Inhibition of tumor necrosis factor-alpha-induced apoptosis by HO-1 overexpression was reversed by 1H (1,2, 4)oxadiazolo(4,3-a)quinoxalin-1-one, an inhibitor of guanylate cyclase, which is a target enzyme for carbon monoxide. Taken together, our data suggest that the antiapoptotic effect of HO-1 may be mediated via carbon monoxide. PMID- 10666116 TI - Electrical behavior of guinea pig tracheal smooth muscle. AB - Intracellular recordings were taken from the smooth muscle of the guinea pig trachea, and the effects of intrinsic nerve stimulation were examined. Approximately 50% of the cells had stable resting membrane potentials of -50 +/- 1 mV. The remaining cells displayed spontaneous oscillations in membrane potential, which were abolished either by blocking voltage-dependent Ca(2+) channels with nifedipine or by depleting intracellular Ca(2+) stores with ryanodine. In quiescent cells, stimulation with a single impulse evoked an excitatory junction potential (EJP). In 30% of these cells, trains of stimuli evoked an EJP that was followed by oscillations in membrane potential. Transmural nerve stimulation caused an increase in the frequency of spontaneous oscillations. All responses were abolished by the muscarinic-receptor antagonist hyoscine (1 microM). In quiescent cells, nifedipine (1 microM) reduced EJPs by 30%, whereas ryanodine (10 microM) reduced EJPs by 93%. These results suggest that both the release of Ca(2+) from intracellular stores and the influx of Ca(2+) through voltage-dependent Ca(2+) channels are important determinants of spontaneous and nerve-evoked electrical activity of guinea pig tracheal smooth muscle. PMID- 10666117 TI - Prenyltransferase inhibitors block superoxide production by pulmonary vascular smooth muscle. AB - We recently showed that the farnesyltransferase inhibitor FTI-277 blocks interleukin 1beta (IL-1beta)-induced nitric oxide production in pulmonary vascular smooth muscle cells (SMC), whereas the geranylgeranyltransferase inhibitor GGTI-298 enhances this effect. Here we show that IL-1beta and platelet derived growth factor (PDGF) stimulate superoxide production by pulmonary vascular SMC and that this effect is blocked by both FTI-277 and GGTI-298, suggesting that farnesylated and geranylgeranylated proteins are required for superoxide production. We also show that FTI-277 and GGTI-298 block superoxide production stimulated by constitutively active mutant H-Ras. Furthermore, superoxide production by IL-1beta, PDGF factor, and constitutively activated Ras is blocked by diphenyleneiodonium, implicating NAD(P)H oxidase as the generating enzyme. Given the role of oxidant radicals in vascular reactivity and injury, the action of both FTI-277 and GGTI-298 in suppressing superoxide generation by an inflammatory cytokine as well as by a potent smooth muscle mitogen may be therapeutically useful. PMID- 10666118 TI - Epoxyeicosatrienoic acids constrict isolated pressurized rabbit pulmonary arteries. AB - Little information is available regarding the vasoactive effects of epoxyeicosatrienoic acids (EETs) in the lung. We demonstrate that 5, 6-, 8,9-, 11,12-, and 14,15-EETs contract pressurized rabbit pulmonary arteries in a concentration-dependent manner. Constriction to 5,6-EET methyl ester or 14,15-EET is blocked by indomethacin or ibuprofen (10(-5) M), SQ-29548, endothelial denuding, or submaximal preconstriction with the thromboxane mimetic U-46619. Constriction of pulmonary artery rings to phenylephrine is blunted by treatment with the epoxygenase inhibitor N-methylsulfonyl-6-(2 propargyloxyphenyl)hexanamide. Pulmonary arteries and peripheral lung microsomes metabolize arachidonate to products that comigrate on reverse-phrase HPLC with authentic regioisomers of 5,6-, 8,9-, 11,12-, and 14,15-EETs, but no cyclooxygenase products of EETs could be demonstrated. Proteins of the CYP2B, CYP2E, CYP2J, CYP1A, and CYP2C subfamilies are present in pulmonary artery and peripheral lung microsomes. Constriction of isolated rabbit pulmonary arteries to EETs is nonregioselective and depends on intact endothelium and cyclooxygenase, consistent with the formation of a pressor prostanoid compound. These data raise the possibility that EETs may contribute to regulation of pulmonary vascular tone. PMID- 10666119 TI - Substance P and neurokinin-1 receptor expression by intrinsic airway neurons in the rat. AB - Tachykinins and their receptors are involved in the amplification of inflammation in the airways. We analyzed the expression of preprotachykinin-A (PPT-A) and neurokinin-1 (NK-1) receptor genes by intrinsic airway neurons in the rat. We also tested the hypothesis that PPT-A-encoded peptides released by these neurons fulfill the requisite role of substance P in immune complex injury of the lungs. We found that ganglion neurons in intact and denervated airways or in primary culture coexpress PPT-A and NK-1 receptor mRNAs and their protein products. Denervated ganglia from tracheal xenografts (nu/nu mice) or syngeneic lung grafts had increased PPT-A mRNA contents, suggesting preganglionic regulation. Formation of immune complexes in the airways induced comparable inflammatory injuries in syngeneic lung grafts, which lack peptidergic sensory fibers, and control lungs. The injury was attenuated in both cases by pretreatment with the NK-1 receptor antagonist LY-306740. We conclude that tachykinins released by ganglia act as a paracrine or autocrine signal in the airways and may contribute to NK-1 receptor mediated amplification of immune injury in the lungs. PMID- 10666120 TI - Brief perinatal hypoxia increases severity of pulmonary hypertension after reexposure to hypoxia in infant rats. AB - We hypothesized that disrupted alveolarization and lung vascular growth caused by brief perinatal hypoxia would predispose infant rats to higher risk for developing pulmonary hypertension when reexposed to hypoxia. Pregnant rats were exposed to 11% inspired oxygen fraction (barometric pressure, 410 mmHg; inspired oxygen pressure, 76 mmHg) for 3 days before birth and were maintained in hypoxia for 3 days after birth. Control rats were born and raised in room air. At 2 wk of age, rats from both groups were exposed to hypoxia for 1 wk or kept in room air. We found that brief perinatal hypoxia resulted in a greater increase in right ventricular systolic pressure and higher ratio of right ventricle to left ventricle plus septum weights after reexposure to hypoxia after 2 wk of age. Moreover, perinatal hypoxic rats had decreased radial alveolar counts and reduced pulmonary artery density. We conclude that brief perinatal hypoxia increases the severity of pulmonary hypertension when rats are reexposed to hypoxia. We speculate that disrupted alveolarization and lung vascular growth following brief perinatal hypoxia may increase the risk for severe pulmonary hypertension with exposure to adverse stimuli later in life. PMID- 10666121 TI - Pulmonary-specific expression of SP-D corrects pulmonary lipid accumulation in SP D gene-targeted mice. AB - Targeted disruption of the surfactant protein (SP) D (SP-D) gene caused a marked pulmonary lipoidosis characterized by increased alveolar lung phospholipids, demonstrating a previously unexpected role for SP-D in surfactant homeostasis. In the present study, we tested whether the local production of SP-D in the lung influenced surfactant content in SP-D-deficient [SP-D(-/-)] and SP-D wild-type [SP-D(+/+)] mice. Rat SP-D (rSP-D) was expressed under control of the human SP-C promoter, producing rSP-D, SP-D(+/+) transgenic mice. SP-D content in bronchoalveolar lavage fluid was increased 30- to 50-fold in the rSP-D, SP-D(+/+) mice compared with the SP-D(+/+) parental strain. Lung morphology, phospholipid content, and surfactant protein mRNAs were unaltered by the increased concentration of SP-D. Likewise, the production of endogenous mouse SP-D mRNA was not perturbed by the SP-D transgene. rSP-D, SP-D(+/+) mice were bred to SP-D(-/-) mice to assess whether lung-selective expression of SP-D might correct lipid homeostasis abnormalities in the SP-D(-/-) mice. Selective expression of SP-D in the respiratory epithelium had no adverse effects on lung function, correcting surfactant phospholipid content and decreasing phosphatidylcholine incorporation significantly. SP-D regulates surfactant lipid homeostasis, functioning locally to inhibit surfactant phospholipid incorporation in the lung parenchyma and maintaining alveolar phospholipid content in the alveolus. Marked increases in biologically active tissue and alveolar SP-D do not alter lung morphology, macrophage abundance or structure, or surfactant accumulation. PMID- 10666122 TI - Estradiol improves pulmonary hemodynamics and vascular remodeling in perinatal pulmonary hypertension. AB - Partial ligation of the ductus arteriosus (DA) in the fetal lamb causes sustained elevation of pulmonary vascular resistance (PVR) and hypertensive structural changes in small pulmonary arteries, providing an animal model for persistent pulmonary hypertension of the newborn. Based on its vasodilator and antimitogenic properties in other experimental studies, we hypothesized that estradiol (E(2)) would attenuate the pulmonary vascular structural and hemodynamic changes caused by pulmonary hypertension in utero. To test our hypothesis, we treated chronically instrumented fetal lambs (128 days, term = 147 days) with daily infusions of E(2) (10 microg; E(2) group, n = 6) or saline (control group, n = 5) after partial ligation of the DA. We measured intrauterine pulmonary and systemic artery pressures in both groups throughout the study period. After 8 days, we delivered the study animals by cesarean section to measure their hemodynamic responses to birth-related stimuli. Although pulmonary and systemic arterial pressures were not different in utero, fetal PVR immediately before ventilation was reduced in the E(2)-treated group (2.43 +/- 0.79 vs. 1.48 +/- 0.26 mmHg. ml( 1). min, control vs. E(2), P < 0.05). During the subsequent delivery study, PVR was lower in the E(2)-treated group in response to ventilation with hypoxic gas but was not different between groups with ventilation with 100% O(2). During mechanical ventilation after delivery, arterial partial O(2) pressure was higher in E(2) animals than controls (41 +/- 11 vs. 80 +/- 35 Torr, control vs. E(2), P < 0. 05). Morphometric studies of hypertensive vascular changes revealed that E(2) treatment decreased wall thickness of small pulmonary arteries (59 +/- 1 vs. 48 +/- 1%, control vs. E(2), P < 0.01). We conclude that chronic E(2) treatment in utero attenuates the pulmonary hemodynamic and histological changes caused by DA ligation in fetal lambs. PMID- 10666123 TI - Hepatocyte growth factor is elevated in chronic lung injury and inhibits surfactant metabolism. AB - Adult respiratory distress syndrome may incorporate in its pathogenesis the hyperplastic proliferation of alveolar epithelial type II cells and derangement in synthesis of pulmonary surfactant. Previous studies have demonstrated that hepatocyte growth factor (HGF) in the presence of serum is a potential mitogen for adult type II cells (R. J. Panos, J. S. Rubin, S. A. Aaronson, and R. J. Mason. J. Clin. Invest. 92: 969-977, 1993) and that it is produced by fetal mesenchymal lung cells (J. S. Rubin, A. M.-L. Chan, D. P. Botarro, W. H. Burgess, W. G. Taylor, A. C. Cech, D. W. Hirschfield, J. Wong, T. Miki, P. W. Finch, and S. A. Aaronson. Proc. Natl. Acad. Sci. USA 88: 415-419, 1991). In these studies, we expand on this possible involvement of HGF in chronic lung injury by showing the following. First, normal adult lung fibroblasts transcribe only small amounts of HGF mRNA, but the steady-state levels of this message rise substantially in lung fibroblasts obtained from animals exposed to oxidative stress. Second, inflammatory cytokines produced early in the injury stimulate the transcription of HGF in isolated fibroblasts, providing a plausible mechanism for the increased amounts of HGF seen in vivo. Third, HGF is capable of significantly inhibiting the synthesis and secretion of the phosphatidylcholines of pulmonary surfactant. Fourth, HGF inhibits the rate-limiting enzyme in de novo phosphatidylcholine synthesis, CTP:choline-phosphate cytidylyltransferase (EC 2.7.7.15). Our data indicate that fibroblast-derived HGF could be partially responsible for the changes in surfactant dysfunction seen in adult respiratory distress syndrome, including the decreases seen in surfactant phosphatidylcholines. PMID- 10666124 TI - Maturational differences in hyperoxic AP-1 activation in rat lung. AB - Immature organisms (neonates; <12 h old) have vastly differing responses to hyperoxic injury than adults. A common feature of hyperoxic gene regulation is involvement of activator protein (AP)-1. We evaluated lung AP-1 binding as well as that of the AP-1 subunit proteins c-Fos, c-Jun, phosphorylated c-Jun, Jun B, and Jun D after exposure to >95% O(2) for 3 days. Unlike adults, neonates showed no increased AP-1 binding in hyperoxia despite a high affinity of the AP-1 binding complexes for phosphorylated c-Jun and Jun D as demonstrated by supershift of these antibodies with the AP-1 complexes. Moreover, neonatal lungs exhibited two distinguishable AP-1 binding complexes, whereas adult lungs had one. In neonates, sequential immunoprecipitation revealed that the lower AP-1 complex was composed of proteins from both the Fos and Jun families, whereas the upper complex consisted of Jun family proteins, with predominance of Jun D. In adults, the single AP-1 complex appeared to involve other Fos or non-Fos or non Jun family proteins as well. Neonatal lungs showed a higher level of Jun B and Jun D immunoreactive proteins in both air and hyperoxia compared with those in adult lungs. These results suggest that significant maturational differences in lung AP-1 complexes exist and that these may explain transcriptional differences in hyperoxic gene regulation. PMID- 10666125 TI - Oxygen induction of epithelial Na(+) transport requires heme proteins. AB - Fetal distal lung epithelial (FDLE) cells exposed to a postnatal O(2) concentration of 21% have higher epithelial Na(+) channel (ENaC) mRNA levels and Na(+) transport relative to FDLE cells grown in a fetal O(2) concentration of 3%. To investigate the mechanism of this process, FDLE monolayers were initially cultured in 3% O(2), and then some were switched to a 21% O(2) environment. Incubation of FDLE cells with the iron chelator deferoxamine, CoCl(2), NiCl(2), or an inhibitor of heme synthesis prevented or diminished the O(2) induction of amiloride-sensitive short-circuit current in FDLE cells. Similarly, defer- oxamine and cobalt prevented O(2)-induced ENaC mRNA expression. Exposure of FDLE cells grown under hypoxic conditions to carbon monoxide increased both ENaC mRNA expression and amiloride-sensitive short-circuit current. We therefore concluded that induction of ENaC mRNA expression and amiloride-sensitive Na(+) transport in FDLE cells by a physiological increase in O(2) concentration seen at birth requires iron and heme proteins. PMID- 10666126 TI - Hypoxia induces hexokinase II gene expression in human lung cell line A549. AB - During adaptation to hypoxic and hyperoxic conditions, the genes involved in glucose metabolism are upregulated. To probe involvement of the transcription factor hypoxia-induced factor-1 (HIF-1) in hexokinase (HK) II expression in human pulmonary cells, A549 cells and small-airway epithelial cells (SAECs) were exposed to stimuli such as hypoxia, deferoxamine (DFO), and metal ions. The largest increase in HK-II (20-fold for mRNA and 2.5-fold for enzymatic activity) was observed in A549 cells when exposed to DFO. All stimuli selectively increased the 5.5-kb rather than 4-kb transcript in A549 cells. Cycloheximide and actinomycin D inhibited these responses. In addition, cells were transfected with luciferase reporter constructs driven by the full-length HK-II 5'-regulatory region (4.0 kb) or various deletions of that region. A549 cells transfected with the 4.0-kb construct and exposed to hypoxia or DFO increased their luciferase activity 7- and 10-fold, respectively, indicating that HK-II induction is, at least in part, due to increased gene transcription. Sixty percent of the inducible activity of the 4.0-kb construct was shown to reside within the proximal 0.5 kb. Additionally, cotransfection with a stable HIF-1 mutant and the 4.0-kb promoter construct resulted in increased luciferase activity under normoxic conditions. These results strongly suggest that HK-II is selectively regulated in pulmonary cells by a HIF-1-dependent mechanism. PMID- 10666127 TI - Gastrointestinal osmoreceptors and renal sodium excretion in humans. AB - The hypothesis that natriuresis can be induced by stimulation of gastrointestinal osmoreceptors was tested in eight supine subjects on constant sodium intake (150 mmol NaCl/day). A sodium load equivalent to the amount contained in 10% of measured extracellular volume was administered by a nasogastric tube as isotonic or hypertonic saline (850 mM). In additional experiments, salt loading was replaced by oral water loading (3.5% of total body water). Plasma sodium concentration increased after hypertonic saline (+3.1 +/- 0.7 mM), decreased after water loading (-3.8 +/- 0.8 mM), and remained unchanged after isotonic saline. Oncotic pressure decreased by 9.4 +/- 1.2, 3.7 +/- 1.2, and 10.7 +/- 1.3%, respectively. Isotonic saline induced an increase in renal sodium excretion (104 +/- 15 to 406 +/- 39 micromol/min) that was larger than seen with hypertonic saline (85 +/- 15 to 325 +/- 39 micromol/min) and water loading (88 +/- 11 to 304 +/- 28 micromol/min). Plasma ANG II decreased to 22 +/- 6, 35 +/- 6, and 47 +/- 5% of baseline after isotonic saline, hypertonic saline, and water loading, respectively. Plasma atrial natriuretic peptide (ANP) concentrations and urinary excretion rates of endothelin-1 were unchanged. In conclusion, stimulation of osmoreceptors by intragastric infusion of hypertonic saline is not an important natriuretic stimulus in sodium-replete subjects. The natriuresis after intragastric salt loading was independent of ANP but can be explained by inhibition of the renin-angiotensin system. PMID- 10666128 TI - Pregnancy-associated reduction in vascular protein kinase C activity rebounds during inhibition of NO synthesis. AB - Vascular reactivity has been shown to be reduced during pregnancy and to be enhanced during chronic inhibition of nitric oxide (NO) synthesis in pregnant rats; however, the cellular mechanisms involved are unclear. The purpose of this study was to investigate whether the pregnancy-induced changes in vascular reactivity are associated with changes in the amount and/or activity of vascular protein kinase C (PKC). Active stress as well as the amount and activity of PKC was measured in deendothelialized thoracic aortic strips from virgin and pregnant rats untreated or treated with the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). In virgin rats, the PKC activator phorbol 12,13-dibutyrate (PDBu, 10(-6) M) and the alpha-adrenergic agonist phenylephrine (Phe, 10(-5) M) caused significant increases in active stress and PKC activity that were inhibited by the PKC inhibitors staurosporine and calphostin C. Western blot analysis in aortic strips of virgin rats showed significant amount of the alpha PKC isoform. Both PDBu and Phe caused significant translocation of alpha-PKC from the cytosolic to the particulate fraction. Compared with virgin rats, the PDBu- and Phe-stimulated active stress and PKC activity as well as the amount and the PDBu- and Phe-induced translocation of alpha-PKC were significantly reduced in late pregnant rats but significantly enhanced in pregnant rats treated with L NAME. The PDBu- and Phe-induced changes in active stress and the amount, distribution, and activity of alpha-PKC in virgin rats treated with L-NAME were not significantly different from that in virgin rats, whereas the changes in pregnant rats treated with L-NAME + the NO synthase substrate L-arginine were not significantly different from that in pregnant rats. These results provide evidence that a PKC-mediated contractile pathway in vascular smooth muscle is reduced during pregnancy and significantly enhanced during chronic inhibition of NO synthesis. The results suggest that one possible mechanism of the pregnancy associated changes in vascular reactivity may involve changes in the amount and activity of the alpha-PKC isoform. PMID- 10666129 TI - Accelerated contractile function and improved fatigue resistance of calf muscles in newborn piglets with IUGR. AB - Asymmetrical intrauterine growth restriction is denoted by disproportional reduction of muscle mass compared with body weight reduction. However, effects on contractile function or tissue development of skeletal muscles were not studied until now. Therefore, isometric force output of serial-stimulated hindlimb plantar flexors was measured in thiopental-anesthetized normal weight (NW) and intrauterine growth-restricted (IUGR) 1-day-old piglets under conditions of normal, reduced (aortic cross clamping), and reestablished (clamp release) blood supply (measured by colored microspheres technique). Furthermore, muscle fiber type distribution was determined after histochemical staining, specific muscle force of the plantar flexors [quotient from absolute force divided by muscle mass (N/g)] was calculated, and glycogen content and morphometric data of the investigated muscles were estimated. Regional blood flow of hindlimb muscles was similar in NW (6 +/- 2 ml. min(-1). 100 g(-1)) and IUGR piglets (8 +/- 1 ml. min( 1). 100 g(-1)). Isometric muscle contractions induced a marked increase in regional blood flow of 4.1-fold in NW and 5-fold in stimulated hindlimb muscles of IUGR piglets (baseline blood flow). Specific force of NW piglet muscles (5.2 +/- 0.2 N/g) was significantly lower than IUGR piglet muscles (6.1 +/- 0.6 N/g; P < 0.05). Isometric muscle contractions (NW: 32.7 +/- 4.7 N; IUGR: 21.7 +/- 4.0 N) resulted in a higher rate of force decrease in the calf muscles of NW animals compared with IUGR piglets (8 +/- 2 vs. 3 +/- 1%; P < 0. 01). Functional restoration of contractile performance after hindlimb recirculation was nearly complete in IUGR piglets (98 +/- 1%), whereas in NW piglets a deficit of 9 +/- 3% was found (P < 0. 01). Muscle fiber type estimation revealed an increased proportion of type I fibers in flexor digitalis superficialis and gastrocnemius medialis in IUGR piglets (P < 0.05). These data clearly indicate that contractile function is accelerated in newborn IUGR piglets. PMID- 10666130 TI - Influence of pontine A5 region on renal sympathetic nerve activity in conscious rabbits. AB - The effects of inhibiting the neural activity in the pontine A5 region on renal sympathetic responses to baroreflex and/or chemoreflex activation were examined in conscious rabbits. Eight rabbits were chronically instrumented with guide cannulas for bilateral microinjections into the A5 area and an electrode for measuring renal sympathetic nerve activity (RSNA). Baroreflex curves were obtained under conditions of normoxia and hypoxia (10% O(2) + 3% CO(2)) after injections into the A5 region of the GABA receptor agonist muscimol or vehicle solution. Under normoxia, injections of muscimol did not affect resting RSNA or blood pressure but increased the range of the RSNA baroreflex by 24 and 33% at doses of 175 or 875 pmol, respectively, without affecting the reflex gain. Hypoxia alone increased resting RSNA by 63%, as well as the range and gain of the RSNA baroreflex by 53 and 89%, respectively, without affecting blood pressure. However, under hypoxia, muscimol increased resting RSNA by 37 and 47% but decreased the gain of the RSNA baroreflex by 19 and 34% at doses of 175 or 875 pmol, respectively, without affecting the reflex range. The effects of muscimol on RSNA were mediated via changes in the amplitude of the sympathetic bursts, whereas burst frequency remained unaffected. These data suggest that the A5 region has a little tonic influence on RSNA in conscious rabbits but serves to limit the renal sympathetic responses to baroreceptor unloading or chemoreceptor stimulation. The different changes in the baroreflex range and gain evoked by muscimol under normoxia and hypoxia indicate that the A5 modulatory action may depend on the activity of the afferent inputs to this region. PMID- 10666131 TI - Spontaneous mutation in the db gene results in obesity and diabetes in CD-1 outbred mice. AB - Five allelic mutants of the diabetes (db) gene have been previously described in mice and rats causing obesity, infertility, and varying degrees of diabetes. We have identified a new, spontaneous mutation resulting in obesity and diabetes in a colony of CD-1 outbred mice, Mus musculus domesticus. Genetic complementation studies indicated that the new mutation was an allele of the diabetes locus. Sequence analysis of cDNA fragments showed a deletion of one G residue located in exon 12 of the leptin receptor gene. The mutation, Lepr(db-NCSU), results in a frameshift and reduces Lepr transcript levels 10-fold. Mutant mice drank up to four times more water and were up to two times heavier than wild-type mice. Blood glucose and plasma insulin and leptin concentrations were sexually dimorphic among affected mice, suggesting an effect of sex steroids. Mortality of affected males was 100% by 5 mo, whereas affected females survived up to 10 mo of age. PMID- 10666132 TI - Effects of vagotomy on serum endotoxin, cytokines, and corticosterone after intraperitoneal lipopolysaccharide. AB - The vagus nerve appears to play a role in communicating cytokine signals to the central nervous system, but the exact extent of its involvement in cytokine-to brain communication remains controversial. Recently, subdiaphragmatic vagotomy was shown to increase bacterial translocation across the gut barrier and thus may cause endotoxin tolerance. The current experiment tested whether or not vagotomized animals have similar systemic responses to endotoxin challenge as do sham-operated animals. Subdiaphragmatically vagotomized and sham-operated animals were injected intraperitoneally with one of three doses (10, 50, 100 microg/kg) of lipopolysaccharide (LPS) or vehicle, and blood samples were taken at 15, 30, 60, 90, and 120 min after the injection. The intraperitoneal injection of LPS increased circulating LPS levels at all time points examined. In addition, all three doses of LPS significantly increased serum interleukin (IL)-1beta, IL-6, and corticosterone in both control and vagotomized rats. In conclusion, vagotomy itself has no marked effect on circulating endotoxin levels or the production of IL-1beta, IL-6, or corticosterone in blood after an intraperitoneal injection of LPS. PMID- 10666133 TI - Angiotensin II indirectly vasoconstricts the ovine uterine circulation. AB - The uterine vasculature of women and sheep predominantly expresses type 2 ANG II receptors that do not mediate vasoconstriction. Although systemic ANG II infusions increase uterine vascular resistance (UVR), this could reflect indirect mechanisms. Thus we compared systemic and local intra-arterial ANG II infusions in six near-term pregnant and five ovariectomized nonpregnant ewes to determine how ANG II increases UVR. Systemic ANG II dose-dependently (P > 0.001) increased arterial pressure (MAP) and UVR and decreased uterine blood flow (UBF) in pregnant and nonpregnant ewes; however, nonpregnant responses exceeded pregnant (P < 0.001). In contrast, local ANG II infusions at rates designed to achieve concentrations in the uterine circulation comparable to those seen during systemic infusions did not significantly decrease UBF in either group, and changes in MAP and UVR were absent or markedly attenuated. When MAP rose during local ANG II, which only occurred with doses > or =2 ng/ml, increases in MAP were delayed more than twofold compared with responses during systemic ANG II infusions and always preceded decreases in UBF, resembling that observed during systemic ANG II infusions. These observations demonstrate attenuated uterine vascular responses to systemic ANG II during pregnancy and suggest that systemic ANG II may increase UVR through release of another potent vasoconstrictor(s) into the systemic circulation. PMID- 10666134 TI - Reduction of food intake by intestinal macronutrient infusion is not reversed by NMDA receptor blockade. AB - Rats increase their intake of food, but not water, after intraperitoneal injection of MK-801, a noncompetitive antagonist of N-methyl-D-aspartate activated ion channels. We hypothesized that MK-801 might enhance intake by interfering with intestinal chemosensory signals. To test this hypothesis, we examined the effect of the antagonist on 15% sucrose intake after an intraduodenal infusion of maltotriose, oleic acid, or phenylalanine in both real- and sham-feeding paradigms. MK-801 (100 microg/kg) significantly increased sucrose intake regardless of the composition of the infusate during real feeding. Furthermore, MK-801 had no effect on reduction of sucrose intake by intestinal nutrient infusions in sham-feeding rats. These results indicate that MK-801 does not increase meal size and duration by interfering with signals activated by intestinal macronutrients. PMID- 10666135 TI - Preinduction of heat shock proteins protects cardiac and hepatic functions following trauma and hemorrhage. AB - Although studies have shown that induction of the heat shock proteins (HSPs), such as HSP-70, has various beneficial effects after ischemia-reperfusion, it remains unknown whether prior induction of HSP-70 has any salutary effects on cardiovascular and hepatocellular functions after trauma-hemorrhage and resuscitation. Male rats were exposed to heat stress (41 degrees C, 15 min) and then allowed to recover for 24 h at room temperature (21 degrees C). The rats then underwent laparotomy (i.e., trauma induced) and were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of the maximal shed blood volume was returned in the form of Ringer lactate. Animals were then resuscitated with four times the volume of shed blood with Ringer lactate over 60 min. The maximal rate of the left ventricular pressure increase or decrease was measured up to 4 h after resuscitation. Cardiac output, hepatocellular function, plasma levels of tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) were determined at 4 h after resuscitation. Cardiac and hepatic tissue were examined for HSP-70 by Western blot analysis. Left ventricular performance, cardiac output, and hepatocellular function decreased significantly following trauma-hemorrhage. Plasma levels of TNF-alpha and IL-6 were also significantly increased. However, prior heat stress attenuated cardiovascular and hepatocellular dysfunction, decreased circulating levels of proinflammatory cytokines following trauma hemorrhage, and was associated with an increased abundance of HSP-70 in the heart and liver. Our data, therefore, suggest that preinduction of HSP-70 protects cardiovascular and hepatocellular functions following trauma-hemorrhage and resuscitation. PMID- 10666136 TI - Effects of intraduodenal glucose and fructose on antropyloric motility and appetite in healthy humans. AB - Oral fructose empties from the stomach more rapidly and may suppress food intake more than oral glucose. The purpose of the study was to evaluate the effects of intraduodenal infusions of fructose and glucose on antropyloric motility and appetite. Ten healthy volunteers were given intraduodenal infusions of 25% fructose, 25% glucose, or 0.9% saline (2 ml/min for 90 min). Antropyloric pressures, blood glucose, and plasma insulin, gastric inhibitory peptide (GIP), and glucagon-like peptide-1 (GLP-1) were measured concurrently; a buffet meal was offered at the end of the infusion. Intraduodenal fructose and glucose suppressed antral waves (P < 0. 0005 for both), stimulated isolated pyloric pressure waves (P < 0.05 for both), and increased basal pyloric pressure (P = 0.10 and P < 0. 05, respectively) compared with saline, without any significant difference between them. Intraduodenal glucose increased blood glucose (P < 0.0005), as well as plasma insulin (P < 0.0005) and GIP (P < 0.005) more than intraduodenal fructose, whereas there was no difference in the GLP-1 response. Intraduodenal fructose suppressed food intake compared with saline (P < 0.05) and glucose (P = 0.07). We conclude that, when infused intraduodenally at 2 kcal/min for 90 min 1) fructose and glucose have comparable effects on antropyloric pressures, 2) fructose tends to suppress food intake more than glucose, despite similar GLP-1 and less GIP release, and 3) GIP, rather than GLP-1, probably accounts for the greater insulin response to glucose than fructose. PMID- 10666137 TI - Stress alters cutaneous permeability barrier homeostasis. AB - Recent studies have shown that psychological stress can influence cutaneous barrier function, suggesting that this form of stress could trigger or aggravate skin disease. In the present study, we demonstrate that transfer of hairless mice to a different cage delays barrier recovery rates. Pretreatment with a phenothiazine sedative, chlorpromazine, before transfer of animals restored the kinetics of barrier recovery toward normal, suggesting that psychological stress is the basis for this alteration in barrier homeostasis. To determine the mechanism linking psychological stress to altered barrier recovery, we first demonstrated that plasma corticosterone levels increase markedly after transfer of animals to new cages and that pretreatment with chlorpromazine blocks this increase. Second, we demonstrated that the systemic administration of corticosterone delays barrier recovery. Finally, we demonstrated that pretreatment with the glucocorticoid receptor antagonist RU-486 blocks the delay in barrier recovery produced by systemic corticosterone, change of cage, or immobilization. These results suggest that psychological stress stimulates increased production of glucocorticoids, which, in turn, adversely affects permeability barrier homeostasis. PMID- 10666138 TI - Daytime naps in darkness phase shift the human circadian rhythms of melatonin and thyrotropin secretion. AB - To systematically determine the effects of daytime exposure to sleep in darkness on human circadian phase, four groups of subjects participated in 4-day studies involving either no nap (control), a morning nap (0900-1500), an afternoon nap (1400-2000), or an evening nap (1900-0100) in darkness. Except during the scheduled sleep/dark periods, subjects remained awake under constant conditions, i.e., constant dim light exposure (36 lx), recumbence, and caloric intake. Blood samples were collected at 20-min intervals for 64 h to determine the onsets of nocturnal melatonin and thyrotropin secretion as markers of circadian phase before and after stimulus exposure. Sleep was polygraphically recorded. Exposure to sleep and darkness in the morning resulted in phase delays, whereas exposure in the evening resulted in phase advances relative to controls. Afternoon naps did not change circadian phase. These findings indicate that human circadian phase is dependent on the timing of darkness and/or sleep exposure and that strategies to treat circadian misalignment should consider not only the timing and intensity of light, but also the timing of darkness and/or sleep. PMID- 10666139 TI - Type of negative feedback controlling sucrose ingestion depends on sucrose concentration. AB - The sham intake of concentrated, but not weak, milk solutions requires up to three sham-feeding tests for intake to reach maximum (7). It is well known that the sham intake of concentrated (0.8 M) sucrose requires three or more sham feeding tests to reach its maximum (4, 6, 17), but it is not known if this occurs with weaker sucrose solutions. We investigated this question by measuring the sham intake of seven concentrations of sucrose (0.025, 0.05, 0.1, 0. 2, 0.4, 0.6, and 0.8 M) during five sham-feeding tests. Sham intake of the three highest concentrations required up to three sham-feeding tests to reach maximum. Sham intake of the four lowest concentrations reached maximum in the first sham feeding test. Our results show that the type of negative feedback that controls the intake of sucrose depends on its concentration. With weak solutions, intake is limited by a single direct, physiological, negative-feedback signal. When concentrated solutions are ingested, intake is controlled jointly by a direct physiological signal operating in conjunction with a labile one that loses its effectiveness with sham-feeding experience. PMID- 10666140 TI - Dexfenfluramine and norfenfluramine: comparison of mechanism of action in feeding and brain Fos-ir studies. AB - Dexfenfluramine (dF) and dexnorfenfluramine (dNF), its metabolite, are anorectic agents that release serotonin (5-HT) and may have a direct postsynaptic action. The effects on the anorectic effects of dF and dNF of either acute (p chlorophenylalanine, PCPA) or chronic (5,7-dihydroxytryptamine, 5,7-DHT) brain 5 HT depletions were studied in rats and compared with the actions of a 5-HT uptake inhibitor (fluoxetine) and 5-HT(1B/2C) receptor agonists [1-(3-trifluoromethyl phenyl)-piperazine and 1-(3-chlorophenyl) piperazine]. The anorexia caused by these agonists was enhanced in rats with 5,7-DHT lesions, possibly a result of receptor supersensitivity. In contrast, fluoxetine anorexia was somewhat reduced in one study and was unchanged in a second. Both dF and dNF anorexias were enhanced in rats with 5,7-DHT lesions. In contrast, the anorectic effects of either dF or dNF were unchanged in PCPA-treated rats relative to controls. Compared with controls, 5, 7-DHT-lesion rats showed greatly increased dF- and dNF induced Fos-like immunoreactivity (ir) in the paraventricular (PVN) and supraoptic (SON) hypothalamic nuclei, and in the median preoptic area (MnPO), but were similar to controls in most other areas. PCPA pretreatment increased dF- and dNF-induced Fos-ir in the PVN, SON, and MnPO. In controls, equianorectic doses of dF and dNF induced Fos-ir in similar brain regions, but dNF produced relatively larger effects than dF in SON, PVN, and MnPO. The data are discussed in terms of multiple pathways in the anorectic actions of dF and dNF. PMID- 10666141 TI - Interstitial K(+) in human skeletal muscle during and after dynamic graded exercise determined by microdialysis. AB - Interstitial K(+) concentrations were measured during one-legged knee-extensor exercise by use of microdialysis with probes inserted in the vastus lateralis muscle of the subjects. K(+) in the dialysate was measured either by flame photometry or a K(+)-sensitive electrode placed in the perfusion outlet. The correction for fractional K(+) recovery was based on the assumption of identical fractional thallium loss. The interstitial K(+) was 4. 19 +/- 0.09 mM at rest and increased to 6.17 +/- 0.19, 7.48 +/- 1.18, and 9.04 +/- 0.74 mM at 10, 30, and 50 W exercise, respectively. The individual probes demonstrated large variations in interstitial K(+), and values >10 mM were obtained. The observed interstitial K(+) was markedly higher than previously found for venous K(+) concentrations at similar work intensities. The present data support a potential role for interstitial K(+) in regulation of blood flow and development of fatigue. PMID- 10666142 TI - Sodium intake is increased by social stress and the Y chromosome and reduced by clonidine. AB - The objectives were to determine 1) if female rats have higher Na intake than males and if social stress increases Na intake, 2) if the sympathetic nervous system (SNS) mediates the stress effects and the gender effect, and 3) if the Y chromosome (Yc) from a hypertensive father increases Na intake. Four rat strains (n = 10/group) of both sexes were used: 1) Wistar Kyoto normotensive (WKY), 2) an F(16) backcross with a Yc from a hypertensive father (SHR/y), 3) spontaneously hypertensive rat (SHR), and 4) an F(16) backcross with a Yc from a normotensive father (SHR/a). Females showed greater baseline Na intake than males (hypertensive strains), intruder stress increased Na intake, and clonidine decreased Na intake, but not in WKY or SHR females. SHR/y males had higher baseline Na intake compared with WKY males. In conclusion, the higher Na intake in females during baseline and stress was partially mediated through the SNS in hypertensive strains and the SHR Yc was partially responsible for the increased Na intake in SHR/y and SHR males compared with WKY. PMID- 10666143 TI - Aminopeptidase-A. I. CDNA cloning and expression and localization in rat tissues. AB - Aminopeptidase-A (APA) is an ectoenzyme that selectively hydrolyzes acidic residues from the amino terminus of oligopeptides, including biologically active [Asp(1)]ANG II and [Asp(1)]CCK-8. We sought to characterize rat APA by cDNA cloning and expression and to determine its tissue distribution by in situ hybridization and immunohistochemistry. Sequence analysis of overlapping cDNA clones isolated from rat kidney cDNA libraries indicated that the full-length cDNA encoded a 945-amino acid protein with a predicted molecular mass of 108 kDa; the size was confirmed by in vitro translation of a full-length cDNA construct. Transient transfection of the full-length cDNA construct in mammalian cells yielded a protein approximately 140 kDa in size, a size that agrees with the immunoblots of APA from rat tissue and is consistent with APA being known as a glycosylated protein. Tissue APA activity and mRNA expression were highest in the kidney and ileum. Localization of APA by in situ hybridization and immunohistochemistry indicated that, with the exception of the kidney and ileum, where APA was localized to the luminal brush border of proximal tubules and enterocytes, respectively, APA was associated with either capillaries or the lining of sinusoids. Areas known to be physiological targets for ANG II, including glomeruli, the zona glomerulosa, and anterior pituitary, had high levels of APA. The localization pattern suggests that APA may subserve multiple functions, i.e., a generalized role in peptide scavenging and perhaps a more specific role in metabolism of circulating or locally produced ANG II or CCK-8. PMID- 10666144 TI - Aminopeptidase-A. II. Genomic cloning and characterization of the rat promoter. AB - Aminopeptidase-A (APA) has a widespread tissue distribution consistent with a role in the metabolism of circulating or locally produced ANG II or CCK-8. APA is also highly expressed in pre-B lymphocytes, but its role in lymphoid cell development is unknown. To begin to understand the basis for cell-specific regulation of APA expression, we sought to clone and characterize the rat gene promoter. Screening of a rat genomic library with a partial rat APA cDNA resulted in isolation of a 12-kb clone found to contain the first exon and >3 kb of 5' flanking sequence. Primer extension of rat kidney mRNA indicated that the major transcription start site was 312 bp upstream of the translation start codon and 22 bp downstream from a TATA box. Constructs containing portions of the 5' flanking region placed upstream of a chloramphenicol acetyltransferase reporter gene indicated that expression was cell specific and that high activity could be obtained with constructs containing as little as 110 bp of 5'-flanking region sequence. We further identified an upstream regulatory element between -1063 and 348 that suppressed transcription in a cell-specific manner. This element (termed upstream suppressor of APA, or USA) also suppressed transcription of a heterologous promoter. These results indicate that the organization and regulation of the rat APA is not consistent with it being a housekeeping gene and further suggest that rat APA gene transcription might be regulated through the presence of a novel strong upstream suppressor element. PMID- 10666145 TI - Chronic infusion of norepinephrine into the VMH of normal rats induces the obese glucose-intolerant state. AB - Increases in ventromedial hypothalamic (VMH) norepinephrine (NE) levels and/or activities have been observed in a variety of animal models of the obese insulin resistant condition. This study examined the metabolic effects of chronic NE infusion (25 nmol/h) into the unilateral VMH of normal rats. Within 4 days, VMH NE infusion significantly increased plasma insulin (140%), glucagon (45%), leptin (300%), triglyceride (100%), abdominal fat pad weight (50%), and white adipocyte lipogenic (100%) and lipolytic (100%) activities relative to vehicle-infused rats. Furthermore, isolated islet insulin secretory response to glucose (15 mM) within 4 days of such treatment was increased over twofold (P < 0.05). Among treated animals, fat stores continued to increase over time and plateaued at approximately 2 wk (3-fold increase), remaining elevated to the end of the study (5 wk). By week 4 of treatment, NE infusion induced glucose intolerance as evidenced by a 32% increase in plasma glucose total area under the glucose tolerance test curve (P < 0.01). Whole body fat oxidation rate measured after 5 wk of infusion was significantly increased among treated animals as evidenced by a reduced respiratory quotient (0.87 +/- 0.01) relative to controls (0. 90 +/- 0.01). VMH NE infusion induced hyperphagia (30%) only during the first week and did not affect body weight over the 5-wk period. Increases in VMH NE activity that are common among obese insulin-resistant animal models can cause the development of this obese glucose-intolerant (metabolic) syndrome. PMID- 10666146 TI - Baroreflex control of muscle sympathetic nerve activity after 120 days of 6 degrees head-down bed rest. AB - To examine how long-lasting microgravity simulated by 6 degrees head-down bed rest (HDBR) induces changes in the baroreflex control of muscle sympathetic nerve activity (MSNA) at rest and changes in responses of MSNA to orthostasis, six healthy male volunteers (range 26-42 yr) participated in Valsalva maneuver and head-up tilt (HUT) tests before and after 120 days of HDBR. MSNA was measured directly using a microneurographic technique. After long-term HDBR, resting supine MSNA and heart rate were augmented. The baroreflex slopes for MSNA during Valsalva maneuver (in supine position) and during 60 degrees HUT test, determined by least-squares linear regression analysis, were significantly steeper after than before HDBR, whereas the baroreflex slopes for R-R interval were significantly flatter after HDBR. The increase in MSNA from supine to 60 degrees HUT was not different between before and after HDBR, but mean blood pressure decreased in 60 degrees HUT after HDBR. In conclusion, the baroreflex control of MSNA was augmented, whereas the same reflex control of R-R interval was attenuated after 120 days of HDBR. PMID- 10666147 TI - Effects of a unique conjugate of alpha-lipoic acid and gamma-linolenic acid on insulin action in obese Zucker rats. AB - The purpose of this study was to assess the individual and interactive effects of the antioxidant alpha-lipoic acid (LPA) and the n-6 essential fatty acid gamma linolenic acid (GLA) on insulin action in insulin-resistant obese Zucker rats. LPA, GLA, and a unique conjugate consisting of equimolar parts of LPA and GLA (LPA-GLA) were administered for 14 days at 10, 30, or 50 mg. kg body wt(-1). day( 1). Whereas LPA was without effect at 10 mg/kg, at 30 and 50 mg/kg it elicited 23% reductions (P < 0.05) in the glucose-insulin index (the product of glucose and insulin areas under the curve during an oral glucose tolerance test and an index of peripheral insulin action) that were associated with significant increases in insulin-mediated (2 mU/ml) glucose transport activity in isolated epitrochlearis (63-65%) and soleus (33-41%) muscles. GLA at 10 and 30 mg/kg caused 21-25% reductions in the glucose-insulin index and 23-35% improvements in insulin-mediated glucose transport in epitrochlearis muscle. The beneficial effects of GLA disappeared at 50 mg/kg. At 10 and 30 mg/kg, the LPA-GLA conjugate elicited 29 and 38% reductions in the glucose-insulin index. These LPA-GLA induced improvements in whole body insulin action were accompanied by 28-63 and 38-57% increases in insulin-mediated glucose transport in epitrochlearis and soleus muscles and resulted from the additive effects of LPA and GLA. At 50 mg/kg, the metabolic improvements due to LPA-GLA were substantially reduced. In summary, these results indicate that the conjugate of the antioxidant LPA and the n-6 essential fatty acid GLA elicits significant dose-dependent improvements in whole body and skeletal muscle insulin action on glucose disposal in insulin resistant obese Zucker rats. Moreover, these actions of LPA-GLA are due to the additive effects of its individual components. PMID- 10666148 TI - Effects of endothelin-1 and homologous trout endothelin on cardiovascular function in rainbow trout. AB - The cardiovascular effects of endothelin (ET)-1 and the recently sequenced homologous trout ET were examined in unanesthetized trout, and vascular capacitance curves were constructed to evaluate the responsiveness of the venous system to ET-1. A bolus dose of 667 pmol/kg ET-1 doubled ventral aortic pressure; produced a triphasic pressor-depressor-pressor response in dorsal aortic pressure (P(DA)); increased central venous pressure, gill resistance, and systemic resistance; and decreased cardiac output, heart rate, and stroke volume. These responses were dose dependent. Bolus injection of trout ET (333 or 1,000 pmol/kg) produced essentially identical, dose-dependent cardiovascular responses as ET-1. Dorsal aortic infusion of 1 and 3 pmol. kg(-1). min(-1) ET-1 and central venous infusion into the ductus Cuvier of 0.3 and 1 pmol. kg(-1). min(-1) produced similar dose-dependent cardiovascular responses, although the increase in P(DA) became monophasic. The heightened sensitivity to central venous infusion was presumably due to the more immediate exposure of the branchial vasculature to the peptide. Infusion of 1 pmol. kg(-1). min(-1) ET-1 decreased vascular compliance but had no effect on unstressed blood volume. These results show that ETs affect a variety of cardiovascular functions in trout and that branchial vascular resistance and venous compliance are especially sensitive. The multiplicity of effectors stimulated by ET suggests that this peptide was extensively integrated into cardiovascular function early on in vertebrate phylogeny. PMID- 10666149 TI - Arterial baroreceptors control plasma vasopressin responses to graded hypotension in conscious dogs. AB - We studied the role of cardiac and arterial baroreceptors in the reflex control of arginine vasopressin (AVP) and renin secretion during graded hypotension in conscious dogs. The dogs were prepared with Silastic cuffs on the thoracic inferior vena cava and catheters in the pericardial space. Each experiment consisted of a control period followed by four periods of inferior vena caval constriction, during which mean arterial pressure (MAP) was reduced in increments of approximately 10 mmHg. The hormonal responses were measured in five dogs under four treatment conditions: 1) intact, 2) acute cardiac denervation (CD) by intrapericardial infusion of procaine, 3) after sinoaortic denervation (SAD), and 4) during combined SAD+CD. The individual slopes relating MAP to plasma AVP and plasma renin activity (PRA) were used to compare the treatment effects using a 2 x 2 factorial analysis. There was a significant (P < 0.01) effect of SAD on the slope relating plasma AVP to MAP but no effect of CD and no SAD x CD interaction. In contrast, the slope relating PRA and MAP was increased (P < 0.05) by SAD but was not affected by CD. These results support the hypothesis that stimulation of AVP secretion in response to graded hypotension is primarily driven by unloading arterial baroreceptors in the dog. PMID- 10666150 TI - Metabolic control of food intake and estrous cycles in syrian hamsters. I. Plasma insulin and leptin. AB - The "adipostat hypothesis" refers to the idea that circulating hormone concentrations reflect levels of body adiposity and act as signals to control food intake and reproduction. Implicit in the adipostatic hypothesis are the following two assumptions: 1) plasma levels of adipostatic hormones accurately reflect body fat content and 2) decreased plasma concentrations of adipostatic hormones necessarily result in increased food intake and inhibited reproductive processes. The present experiments are designed to test these assumptions. Fat and lean Syrian hamsters were either fasted for 12, 24, 36, or 48 h or allowed ad libitum access to food. Contrary to the first assumption, plasma leptin and insulin levels in fat hamsters dropped dramatically by 12 h after the start of a fast, with no significant change in body fat content and no postfast hyperphagia. Lean hamsters showed anestrus after a 48-h fast but not after a 24-h fast. Contrary to the second assumption of the lipostatic hypothesis, lean hamsters fasted for 24 h and then refed for the next 24 h had leptin levels that were not significantly elevated compared with those of 48-h-fasted hamsters. Thus, in adult female Syrian hamsters, plasma leptin concentrations do not accurately reflect body fat content under all conditions; normal estrous cyclicity does not necessarily require plasma leptin concentrations higher than those of fasted hamsters; and decreased plasma leptin levels do not result in increased food intake. PMID- 10666151 TI - Development of the pulmonary surfactant system in two oviparous vertebrates. AB - In birds and oviparous reptiles, hatching is often a lengthy and exhausting process, which commences with pipping followed by lung clearance and pulmonary ventilation. We examined the composition of pulmonary surfactant in the developing lungs of the chicken, Gallus gallus, and of the bearded dragon, Pogona vitticeps. Lung tissue was collected from chicken embryos at days 14, 16, 18 (prepipped), and 20 (postpipped) of incubation and from 1 day and 3 wk posthatch and adult animals. In chickens, surfactant protein A mRNA was detected using Northern blot analysis in lung tissue at all stages sampled, appearing relatively earlier in development compared with placental mammals. Chickens were lavaged at days 16, 18, and 20 of incubation and 1 day posthatch, whereas bearded dragons were lavaged at day 55, days 57-60 (postpipped), and days 58-61 (posthatched). In both species, total phospholipid (PL) from the lavage increased throughout incubation. Disaturated PL (DSP) was not measurable before 16 days of incubation in the chick embryo nor before 55 days in bearded dragons. However, the percentage of DSP/PL increased markedly throughout late development in both species. Because cholesterol (Chol) remained unchanged, the Chol/PL and Chol/DSP ratios decreased in both species. Thus the Chol and PL components are differentially regulated. The lizard surfactant system develops and matures over a relatively shorter time than that of birds and mammals. This probably reflects the highly precocial nature of hatchling reptiles. PMID- 10666152 TI - Effect of NPY in the hypothalamic paraventricular nucleus on uncoupling proteins 1, 2, and 3 in the rat. AB - Neuropeptide Y (NPY) injected into the hypothalamic paraventricular nucleus (PVN) stimulates feeding and decreases uncoupling protein (UCP)-1 mRNA in brown adipose tissue (BAT). The present studies were undertaken to determine whether UCP-2 in white adipose tissue (WAT) and UCP-3 in muscle are regulated by NPY in the PVN. PVN-cannulated male Sprague-Dawley rats were injected with either saline or NPY (PVN, 117 pmol, 0.5 microl) every 6 h for 24 h. NPY in the PVN stimulated feeding and decreased UCP-1 mRNA in BAT independent of NPY-induced feeding. UCP-2 mRNA in WAT was unchanged by NPY. In acromiotrapezius muscle, NPY decreased UCP-3 mRNA, but this was reversed by restricting food intake to control levels. In biceps femoris muscle, NPY alone had no effect on UCP-3 mRNA, but UCP-3 mRNA was significantly increased in the NPY-treated rats that were restricted to control levels of intake. These results suggest that UCP-2 in WAT and UCP-3 in muscle are not subject to specific regulation by NPY in the PVN. PMID- 10666153 TI - Regional effect of naltrexone in the nucleus of the solitary tract in blockade of NPY-induced feeding. AB - Naltrexone (NLTX) in the nucleus of the solitary tract (NTS) decreases feeding induced by neuropeptide Y (NPY) in the paraventricular nucleus (PVN). We sought to determine the NTS region most sensitive to NLTX blockade of PVN NPY-induced feeding. Male Sprague-Dawley rats were fitted with two cannulas; one in the PVN and one in a hindbrain region: caudal, medial, or rostral NTS or 1 mm outside the NTS. Animals received NLTX (0, 1, 3, 10, and 30 microg in 0.3 microl) into the hindbrain region just prior to PVN NPY (0.5 microg, 0.3 microl) or artificial cerebrospinal fluid (0.3 microl). Food intake was measured at 2 h following injection. PVN NPY stimulated feeding, and NLTX in the medial NTS significantly decreased NPY-induced feeding at 2 h, whereas administration of NLTX in the other hindbrain regions did not significantly influence PVN NPY induced feeding. These data suggest that opioid receptors in the medial NTS are most responsive to feeding signals originating in the PVN after NPY stimulation. PMID- 10666154 TI - Lower calorie intake enhances muscle insulin action and reduces hexosamine levels. AB - Previous studies have demonstrated enhanced insulin sensitivity in calorie restricted [CR, fed 60% ad libitum (AL) one time daily] compared with AL-fed rats. To evaluate the effects of reduced food intake, independent of temporal differences in consumption, we studied AL (unlimited food access)-fed and CR (fed one time daily) rats along with groups temporally matched for feeding [fed 3 meals (M) daily]: MAL and MCR, eating 100 and 60% of AL intake, respectively. Insulin-stimulated glucose transport by isolated muscle was increased in MCR and CR vs. AL and MAL; there was no significant difference for MCR vs. CR or MAL vs. AL. Intramuscular triglyceride concentration, which is inversely related to insulin sensitivity in some conditions, did not differ among groups. Muscle concentration of UDP-N-acetylhexosamines [end products of the hexosamine biosynthetic pathway (HBP)] was lower in MCR vs. MAL despite unaltered glutamine fructose-6-phosphate aminotransferase activity (rate-limiting enzyme for HBP). These results indicate that the CR-induced increase in insulin-stimulated glucose transport in muscle is attributable to an altered amount, not timing, of food intake and is independent of lower triglyceride concentration. They further suggest that enhanced insulin action might involve changes in HBP. PMID- 10666155 TI - Tissue hypoxygenation activates the adrenomedullin system in vivo. AB - Our study aimed to investigate the influence of tissue hypo-oxygenation on the adrenomedullin (ADM) system in vivo. For this purpose, male Sprague-Dawley rats were exposed to normobaric hypoxia (8% oxygen) or to functional anemia [0.1% carbon monoxide (CO)] or to cobalt chloride (60 mg/kg) for 6 h. Messenger RNA levels for ADM and its receptor (ADM-R) were assessed in diverse organs by RNase protection assay. Additionally, ADM protein concentrations in these organs, as in plasma, were determined by a RIA. We found that ADM mRNA abundance increased in response to hypoxia and to CO inhalation up to 15-fold in all organs examined. Similarly, ADM-R mRNA abundance increased during hypoxia and CO inhalation in all organs examined with exception of the liver. The effects of hypoxia and of CO inhalation on ADM and ADM-R mRNAs were mimicked by injection of cobaltous chloride. Hypoxia also significantly increased ADM protein content in all organs, and plasma levels of ADM rose twofold in response to hypoxia and CO inhalation. These findings indicate that tissue hypoxia leads to a widespread activation of the ADM system, which comprises a parallel stimulation of ADM and ADM receptor mRNA as enhanced ADM protein synthesis and secretion. The ADM system may, therefore, play a significant role in the physiological response to tissue hypoxia. It appears that ADM and ADM-R belong to the family of classic oxygen regulated genes, which are activated by a decrease of the pericellular oxygen tension through the same intracellular signaling cascade. PMID- 10666156 TI - AMPA glutamate receptors and respiratory control in the developing rat: anatomic and pharmacological aspects. AB - The developmental role of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) glutamate receptors in respiratory regulation remains undefined. To study this issue, minute ventilation (V(E)) was measured in 5-, 10-, and 15-day-old intact freely behaving rat pups using whole body plethysmography during room air (RA), hypercapnic (5% CO(2)), and hypoxic (10% O(2)) conditions, both before and after administration of the non-N-methyl-D-aspartate (NMDA) receptor antagonist 1,2,3, 4-tetrahydro-6-nitro-2,3-dioxobenzo[f]quinoxaline-7-sulfonamide disodium (NBQX; 10 mg/kg ip). In all age groups, V(E) during RA was unaffected by NBQX, despite reductions in breathing frequency (f) induced by increases in both inspiratory and expiratory duration. During hypoxia and hypercapnia, V(E) increases were similar in both NBQX and control conditions in all age groups. However, tidal volume was greater and f lower after NBQX. To determine if AMPA receptor-positive neurons are recruited during hypoxia, immunostaining for AMPA receptor (GluR2/3) and c-fos colabeling was performed in caudal brain stem sections after exposing rat pups at postnatal ages 2, 5, 10, and 20 days, and adult rats to room air or 10% O(2) for 3 h. GluR2/3 expression increased with postnatal age in the nucleus of the solitary tract (NTS) and hypoglossal nucleus, whereas a biphasic pattern emerged for the nucleus ambiguus (NA). c-fos expression was enhanced by hypoxia at all postnatal ages in the NTS and NA and also demonstrated a clear maturational pattern. However, colocalization of GluR2/3 and c-fos was not affected by hypoxia. We conclude that AMPA glutamate receptor expression in the caudal brain stem is developmentally regulated. Furthermore, the role of non-NMDA receptors in respiratory control of conscious neonatal rats appears to be limited to modest, albeit significant, regulation of breathing pattern. PMID- 10666157 TI - Is fluid-phase endocytosis conserved in hepatocytes of species acclimated and adapted to different temperatures? AB - Our primary objective was to determine if rates of fluid-phase endocytosis (FPE) were conserved in hepatocytes from organisms acclimated and adapted to different temperatures. To this aim, the fluorescent dye Lucifer yellow was employed to measure FPE at different assay temperatures (AT) in hepatocytes from 5 degrees C- and 20 degrees C-acclimated trout, Oncorhynchus mykiss (at 5 and 20 degrees C AT), 22 degrees C- and 35 degrees C-acclimated tilapia, Oreochromis nilotica (at 22 and 35 degrees C AT), and the Sprague-Dawley rat (at 10, 20, and 37 degrees C AT). FPE was also studied in rats fed a long-chain polyunsaturated fatty acid (PUFA)-enriched diet (at 10 degrees C AT). Despite being temperature dependent, endocytic rates (values in pl. cell(-1). h(-1)) in both species of fish were compensated after a period of acclimation. For example, in 20 degrees C acclimated trout, the rate of endocytosis declined from 1.84 to 1.07 when the AT was reduced from 20 to 5 degrees C; however, after a period of acclimation at 5 degrees C, the rate (at 5 degrees C AT) was largely restored (1.80) and almost perfectly compensated (95%). In tilapia, endocytic rates were also temperature compensated, although only partially (36%). Relatively similar rates obtained at 5 degrees C in 5 degrees C-acclimated trout (1.8), at 20 degrees C in 20 degrees C-acclimated trout (1.84), and at 22 degrees C in 22 degrees C-acclimated tilapia (2.2) suggest that endocytic rates are somewhat conserved in these two species of fish. In contrast, the rate in rat measured at 37 degrees C (16.83) was severalfold greater than in fish at their respective body temperatures. A role for lipids in determining rates of endocytosis was supported by data obtained at 10 degrees C in hepatocytes isolated from rats fed a long-chain PUFA-enriched diet: endocytic rates were higher (5.35 pl. cell(-1). h(-1)) than those of rats fed a standard chow diet (2.33 pl. cell(-1). h(-1)). The conservation of endocytic rates in fish may be related to their ability to conserve other membrane characteristics (i.e., order or phase behavior) by restructuring their membrane lipid composition or by modulating the activities of proteins that regulate endocytosis and membrane traffic, whereas the lack of conservation between fish and rat may be due to differences in metabolic rate. PMID- 10666158 TI - Tumor necrosis factor (TNF)-alpha induces leptin production through the p55 TNF receptor. AB - Tumor necrosis factor (TNF)-alpha acts directly on adipocytes to increase production of the lipostatic factor, leptin. However, which TNF receptor (TNFR) mediates this response is not known. To answer this question, leptin was measured in plasma of wild-type (WT), p55, and p75 TNFR knockout (KO) mice injected intraperitoneally with murine TNF-alpha and in supernatants from cultured WT, p55, and p75 TNFR KO adipocytes incubated with TNF-alpha. Leptin also was measured in supernatants from C3H/HeOuJ mouse adipocytes cultured with blocking antibodies to each TNFR and TNF-alpha as well as in supernatants from adipocytes incubated with either human or murine TNF-alpha, which activate either one or both TNFR, respectively. The results using all four strategies show that the induction of leptin production by TNF-alpha requires activation of the p55 TNFR and that although activation of the p75 TNFR alone cannot cause leptin production, its presence affects the capability of TNF-alpha to induce leptin production through the p55 TNFR. These results provide new information on the interplay between cells of the immune system and adipocytes. PMID- 10666159 TI - The bone and joint decade 2000-2010. PMID- 10666160 TI - Crystal unclear. PMID- 10666161 TI - Bridging the gap in rheumatology. PMID- 10666162 TI - Right ventricular diastolic abnormalities in systemic sclerosis. Relation to left ventricular involvement and pulmonary hypertension. AB - OBJECTIVES: To investigate right ventricular diastolic function in systemic sclerosis (SSc) and its relation to clinical features of the disease. METHODS: Seventy seven unselected SSc patients and 33 healthy subjects were submitted to echocardiography and echo Doppler study to assess left and right systolic as well diastolic function and to estimate maximal arterial systolic pulmonary pressure (PAP). In addition, the patients were investigated to define the SSc subset and the extent of skin and internal organ involvement. RESULTS: An abnormal right ventricular filling, as expressed by an inverted tricuspidal (Tr) E/A ratio (Tr E/A ratio <1), was detected in 31 of the 77 SSc patients (40%) and in 0 of the 36 controls ( p<0.001 ). All the 31 patients with an inverted Tr E/A ratio were found to have a PAP > 30 mm Hg. Twenty resulted to have an inverted mitral (Mit) E/A ratio (Mit E/A ratio <1), indicating an abnormal left ventricular filling. In multiple regression analysis, Tr E/A ratio resulted to be independently correlated to both PAP (r= -0.35;p<0. 003) and Mit E/A ratio (r=0.39;p<0.001). CONCLUSIONS: This study points out an impaired right ventricular filling in a significant percentage of SSc patients whatever the subset. This alteration is independently correlated to both PAP and left ventricular filling abnormalities. PMID- 10666163 TI - The clinical relevance of antibodies to ribosomal-P common epitope in two targeted systemic lupus erythematosus populations: a large cohort of consecutive patients and patients with active central nervous system disease. AB - OBJECTIVES: To develop an enzyme linked immunosorbent assay (ELISA) using as substrate a synthetic 22-aminoacid peptide, corresponding to the ribosomal P0, P1 and P2 common epitope. To study the specificity and sensitivity of the method and evaluate the frequency and clinical associations of anti-P antibodies in two groups of systemic lupus erythematosus (SLE) patients: (a) unselected SLE patients and (b) SLE patients with central nervous system (CNS) involvement. PATIENTS AND METHODS: The C-terminal 22 aminoacid peptide of the ribosomal P proteins (Lys-Lys-Glu-Glu-Lys-Lys-Glu-Glu-Lys-Ser-Glu-Glu-Glu-Asp-Glu-Asp-Met- Gly-Phe-Gly-Leu-Phe-Asp) was synthesised according to Merrifield's solid phase procedure. Purification of the peptide was performed by preparative high performance liquid chromatography and confirmed by amino acid analysis. Using this peptide, in a concentration 5 microg/ml, an ELISA was developed. The presence of anti-P antibodies was evaluated by western blot using purified ribosomal proteins from rat liver. Sera from 178 consecutive patients with SLE and 28 patients with SLE and CNS manifestations were tested. Sera from 58 patients with rheumatoid arthritis and 57 patients with primary Sjogren's syndrome were used as controls. The cut off point of the assay was defined using 124 normal sera. RESULTS: The specificity of the assay was evaluated by homologous inhibition. Pretreatment of positive sera with soluble 22mer peptide of the ribosomal P proteins resulted in 88% inhibition. The concordance between the peptide assay and western blot was found to be 83%. Thirty three of 178 (18. 6%) of the unselected SLE patients had antibodies to P-protein common epitope. Their presence was associated with more active disease (European Consensus Lupus Activity Measurement, ECLAM scoring system) (p<0.001), higher levels of anti-ds DNA antibodies (p<0.05) and lower levels of the C4 component of complement (p<0.01). Eleven of 28 (39.3%) patients with SLE and active CNS involvement had antibodies to P-protein. The overall prevalence of anti-P antibodies in active CNS disease patients was statistically significantly higher, as compared with unselected SLE patients (chi(2)=6.04, p<0.05). These antibodies were found in a high proportion of patients without anticardiolipin antibodies (52.4%) and they were associated with diffuse CNS involvement (psychiatric disorders (71%) and epilepsy (75%)). CONCLUSIONS: A synthetic analogue of the common epitope of ribosomal P-proteins can be use as an antigen for the detection of anti-P antibodies. These antibodies are associated with active SLE and CNS involvement particularly in patients without anticardiolipin antibodies. PMID- 10666164 TI - Hormone replacement therapy and patterns of osteoarthritis: baseline data from the Ulm Osteoarthritis Study. AB - OBJECTIVES: It has been suggested that hormone replacement therapy (HRT) may protect against osteoarthritis (OA). The aim of this paper was to assess the association between HRT and radiographically defined patterns of OA. METHODS: 475 consecutive women aged 50 years or older (mean age 66.1) who underwent hip or knee joint replacement because of advanced OA in four hospitals in south west Germany were enrolled in a cross sectional study. Participants underwent a standardised interview including detailed history of medication use and a physical examination. Furthermore, radiographs of the joint being replaced and of the contralateral joint as well as of both hands were obtained. Patients were categorised as having bilateral or unilateral OA according to the presence or absence of radiographic OA in the contralateral joint. If radiographic OA of different hand and finger joint groups was present, participants were categorised as having generalised OA (GOA). Logistic regression was used to estimate odds ratios and their 95% confidence intervals for the association between HRT and bilateral or GOA while adjusting for potential confounders. RESULTS: Fifty five women (11.6%) were using HRT. The median duration of use was 5.4 years. The prevalence of bilateral and GOA was similar among users of ORT (86.3% and 27.5%, respectively) and among non-users of HRT (88.7% and 35.7%, respectively). After adjustment for potential confounding factors, the odds ratios (95% confidence intervals) of bilateral OA and GOA among HRT users compared with non-users was 1.21 (0.48, 3.03) and 1. 21 (0.53, 2.74), respectively. CONCLUSION: Despite limited generalisability because of the selective study sample, these data do not support the hypothesis that HRT acts as a systemic protective factor against OA. PMID- 10666165 TI - Bone mineral density in systemic lupus erythematosus: comparison with rheumatoid arthritis and healthy controls. AB - OBJECTIVES: To examine bone mineral density (BMD) frequency of osteoporosis and reduced bone mass in systemic lupus erythematosus (SLE), and compare the data of the SLE patients with matched rheumatoid arthritis (RA) patients and healthy controls. Secondly, to study possible correlations between BMD, demographic and disease variables in the SLE patients. METHODS: Measures of BMD assessed by dual energy x ray absorptiometry were obtained from 75 SLE patients aged /=0.5 g/day) was taken as onset of renal SLE. Initial univariate analysis to determine which factors are associated with onset of renal disease was followed by using a Cox's proportional hazards regression model enabling analysis of several prognostic factors at the same time. Variables included three measures of socioeconomic status, ethnic group and the presence or absence of different autoantibodies. RESULTS: There was no effect from any socioeconomic variable. Using forwards stepwise selection, the following had independent effects (p<0.05) on the development of renal SLE: Afro-Caribbean race (hazard rate ratio 4.4 (1.9 10.2), compared with white population); and the presence of IgG anti-cardiolipin antibodies (hazard rate ratio 2.6 (1.2-5.7)). CONCLUSION: Differing socioeconomic factors do not explain the increased frequency of lupus nephritis in Afro Caribbean patients with SLE, but rather there are important genetic or other host differences. The independent effect of IgG anti-cardiolipin antibodies warrants further investigation. PMID- 10666167 TI - Transverse myelopathy in systemic lupus erythematosus: an analysis of 14 cases and review of the literature. AB - OBJECTIVE: To give a comprehensive review of transverse myelopathy (TM), a rare but serious condition reported in 1-2% of patients with systemic lupus erythematosus (SLE). METHODS: 14 patients with SLE and TM were evaluated and 91 additional cases published in the English and German literature reviewed. RESULTS: TM presented either as the initial manifestation or within five years of the diagnosis of SLE. Most patients presented with a detectable sensory deficit at the thoracic level. In our 14 patients, 22% of the patients showed complete neurological recovery, whereas in the total patient population of 105 (our cases plus those reviewed in the literature), complete recovery was observed in 50%, partial recovery in 29% and no improvement or deterioration in 21%. Treatment with intravenous methylprednisolone followed by cyclophosphamide seemed to be most effective. Seventy per cent of the total patient population had abnormal magnetic resonance imaging findings. In our group of 14 patients, those with higher disease activity (measured by the SLAM) at onset of TM were treated more aggressively (for example, with plasmapheresis and intravenous pulse cyclophosphamide). TM in our patients was associated with antiphospholipid antibodies in 43% of the cases as compared with 64% of the total patient population. Optic neuritis occurred in 48% of the total patient population with SLE and TM, suggesting an association. CONCLUSIONS: TM in SLE is a poorly understood entity. Outcome might be more favourable than previously suggested. There is an association of TM with antiphospholipid antibodies in SLE patients. Treatment including intravenous cyclophosphamide may improve the final outcome. This report emphasises the need for multicentre trials to establish guidelines for optimal treatment. PMID- 10666168 TI - Angiotensin converting enzyme in human synovium: increased stromal [(125)I]351A binding in rheumatoid arthritis. AB - OBJECTIVE: To determine whether tissue angiotensin converting enzyme (ACE) is increased in synovia from patients with rheumatoid arthritis, osteoarthritis or chondromalacia patellae. METHODS: Sections of synovia from patients with rheumatoid arthritis (n = 7), osteoarthritis (n = 7) or chondromalacia patellae (n = 6) were tested for immunoreactivity for ACE, and for binding of the ACE inhibitor [(125)I]351A. The amount of ACE was measured with computer assisted image analysis as the proportion of synovial section area occupied by ACE immunoreactive cells, and the density of [(125)I]351A binding. RESULTS: [(125)I]351A binding sites had characteristics of ACE and colocalised with ACE like immunoreactivity to microvascular endothelium and fibroblast-like stromal cells in inflamed and non-inflamed human synovium. Stromal [(125)I]351A binding densities (B(eq)) and the fraction of synovial section area occupied by ACE immunoreactivity (fractional area) were higher in synovia from patients with rheumatoid arthritis (B(eq) 28 amol/mm(2), fractional area 0.21) than from those with osteoarthritis (B(eq) 9 amol/mm(2), fractional area 0.10) or chondromalacia patellae (B(eq) 9 amol/mm(2), fractional area 0.09)(p < 0.05). Density of [(125)I]351A binding to stroma was similar to that to blood vessels in rheumatoid arthritis, but less dense than vascular binding in chondromalacia patellae and osteoarthritis. Increases in [(125)I]351A binding densities were attributable to increases in the numbers of binding sites, and were consistent with an increase in the density of ACE bearing stromal cells. CONCLUSION: ACE is upregulated in synovial stroma in rheumatoid arthritis. Increased tissue ACE may result in increased local generation of the vasoconstrictor and mitogenic peptide angiotensin II and thereby potentiate synovial hypoxia and proliferation in rheumatoid arthritis. PMID- 10666169 TI - Soluble thrombomodulin concentration is raised in scleroderma associated pulmonary hypertension. AB - OBJECTIVE: To investigate the expression of thrombomodulin in scleroderma associated pulmonary hypertension. METHODS: Soluble thrombomodulin (sTM), was measured in plasma samples from 34 scleroderma patients shown to have pulmonary hypertension at echocardiogram, and comparison drawn against samples from 38 scleroderma control patients, and 20 healthy controls. Serial measurements of sTM were performed in the 34 patients with scleroderma associated pulmonary hypertension to investigate possible changes in sTM concentration with progression of the condition. RESULTS: Mean sTM was raised in scleroderma associated pulmonary hypertension when compared with scleroderma controls (mean sTM 65.4 ng/ml v 43.3 ng/ml, p<0.05), and when compared with healthy controls (mean sTM 38.1 ng/ml, p<0.05). There was no significant difference between mean sTM in scleroderma controls and healthy controls. Mean sTM concentration did not change with progression of pulmonary hypertension. CONCLUSION: Plasma sTM is raised in scleroderma associated pulmonary hypertension. The pathogenesis of scleroderma associated pulmonary hypertension may be distinct from the pathogenesis of other forms of pulmonary vascular disease. PMID- 10666170 TI - Quantitative analyses of sacroiliac biopsies in spondyloarthropathies: T cells and macrophages predominate in early and active sacroiliitis- cellularity correlates with the degree of enhancement detected by magnetic resonance imaging. AB - OBJECTIVE: Sacroiliitis is a hallmark of the spondyloarthropathies (SpA). The degree of inflammation can be quantified by magnetic resonance imaging (MRI). The aim of this study was to further elucidate the pathogenesis of SpA by quantitative cellular analysis of immunostained sacroiliac biopsy specimens and to compare these findings with the degree of enhancement in the sacroiliac joints (SJ) as detected by dynamic MRI. METHODS: The degree of acute sacroiliitis detected by MRI after intravenous administration of gadolinium-DTPA was quantitatively assessed by calculating the enhancement observed in the SJ and chronic changes were graded as described in 32 patients with ankylosing spondylitis (n=18), undifferentiated SpA (n=12) and psoriatic arthritis (n=2). Back pain was graded on a visual analogue scale (VAS, 0-10) and disease duration (DD) was assessed. Shortly after MRI, SJ of patients with VAS > 5 were biopsied guided by computed tomography. Immunohistological examination was performed using the APAAP technique; only whole sections > 3 mm were counted. RESULTS: By MRI, chronic changes II in 13 patients (group II, DD 7.3 (SD 4.8) years), while enhancement < 70% was found in eight (group A, DD 5.6 (SD 3.3) years) and > 70% in 12 patients (group B, DD 4.7 (SD 5.8) years). The relative percentage of cartilage (78-93%), bone (7-18%) and proliferating connective tissue (1-4%) was comparable between the groups (range). There were more inflammatory cells in group I compared with group II (mean (SD) 26.7(20.1) versus 5.3 (5. 2), p=0.04) and group A compared with B (21.8 (17.3) versus 6.0 (5. 6), p=0.05) cells/10 mm(2)), T cells (10.9 (8.5)) being slightly more frequent than macrophages (9.6 (16.8/10 mm(2))). Clusters of proliferating fibroblasts were seen in three and new vessel formation in seven cases. CONCLUSION: This study shows that T cells and macrophages are the most frequent cells in early and active sacroiliitis in SpA. The correlation of cellularity and MRI enhancement provides further evidence for the role of dynamic MRI to detect early sacroiliitis. PMID- 10666171 TI - The relative prevalence of dermatomyositis and polymyositis in Europe exhibits a latitudinal gradient. PMID- 10666172 TI - Synovial membrane p53 protein immunoreactivity in rheumatoid arthritis patients. AB - OBJECTIVES: To examine the expression of the p53 protein in synovial membrane of rheumatoid arthritis (RA) patients and to compare this with the expression in normal synovial tissues in subjects without RA. METHODS: Immunohistological expression of the p53 protein was studied using a streptavidin-biotin-peroxidase method and the monoclonal antibody DO-7, an antibody directed against both wild and mutant forms of p53 protein, in synovial tissues of RA patients (n=10) and from subjects with no known joint disease (n=4). RESULTS: p53 protein expression was present in a small percentage of synovial cells in the majority of the RA patients (n=8; 80%) and in half of the normal control cases with no inflammatory joint disease (n=2; 50%). No sample had more than 5% cells staining with intranuclear pattern. The difference in synovial p53 immunoreactivity between the RA patients and normal controls is not statistically significant (p= 0.64; chi(2) contingency test). CONCLUSIONS: This study has shown that p53 protein is only weakly expressed in the rheumatoid synovial membrane, with a low percentage of p53 protein immunostaining cells present, with intranuclear staining. These results suggest this is wild type p53 protein rather than mutant protein. These findings suggest that synovial p53 protein expression may not be important in the pathogenesis of RA and may only represent a reactive repair process to DNA damage secondary to the immune and inflammatory reactions associated with the disease. PMID- 10666173 TI - Effects of cyclosporin at various concentrations on dexamethasone intracellular uptake in multidrug resistant cells. AB - BACKGROUND: The multidrug resistance phenomenon results from the expression of P glycoprotein (P-gp), a drug-efflux pump. Corticosteroids are substrates for P-gp, whose function can be inhibited by cyclosporin. This study evaluates the ability of cyclosporin to modulate dexamethasone uptake in multidrug resistant cells. METHODS: The K 562 cell line, which does not express P-gp and a P-gp expressing clone, K562/ADM, were used. Cells were incubated with H3-dexamethasone in the absence or presence of cyclosporin at various concentrations. Then, cells were washed, lysed, and radioactivity was measured. RESULTS: The uptake of dexamethasone alone was higher in sensitive than in resistant cells. Addition of cyclosporin induced a dose dependent increase of dexamethasone uptake in resistant cells, whereas the drug did not influence dexamethasone uptake in parental cells. CONCLUSION: Cyclosporin, at therapeutic concentrations induces a moderate, but significant increase in dexamethasone accumulation in multidrug resistant cells. Thus, cyclosporin might increase the intestinal absorption of corticosteroids or their accumulation in mononuclear cells, or both, thereby increasing their therapeutic efficacy. PMID- 10666174 TI - A prospective study on the incidence of rheumatoid arthritis among people with persistent increase of rheumatoid factor. AB - OBJECTIVES: To study the stability of rheumatoid factor (RF) increases and to compare the incidence of rheumatoid arthritis (RA) in people with transient or persistent increase of one or more RF isotypes. METHODS: From an original cohort of nearly 14 000 participants in a population study, 135 previously RF positive persons were recruited in 1996 and evaluated according to the 1987 ACR criteria. The observation time ranged from 9-22 years (mean 16. 5). Blood samples were obtained from all participants at entry and again in 1996. RESULTS: About 40% of the participants who had only one raised RF isotype in the original sample had become RF negative in 1996 compared with only 15% of those with increase of two or three RF isotypes (p=0.002). The seven participants who developed RA during the study period all had persistently raised RF. Six of the 54 participants with more than one RF isotype raised in 1996 developed RA, corresponding to an annual incidence of 0.67%, which was 7.5 times higher than observed in the other participants (p=0. 045). CONCLUSION: Symptom free persons with persistently raised RF have greatly increased risk of developing RA. This suggests that dysregulation of RF production is a predisposing factor in RA. PMID- 10666175 TI - The existence of geographical clusters of cases of inflammatory polyarthritis in a primary care based register. AB - OBJECTIVES: To determine whether there is any evidence that there are spatial clusters of rheumatoid arthritis in particular, and inflammatory arthritis in general. METHODS: Setting was a population based incidence register of inflammatory arthritis: the Norfolk Arthritis Register (NOAR). All cases identified between 1990-1995 were mapped to place of residence. Statistical evidence of clustering was determined by calculating Poisson probabilities in putative areas. RESULTS: Three clusters were identified including one small area (population 85) where five unrelated cases developed during this time period. There was no obvious greater disease homogeneity within clusters and no common environmental factors were identified. CONCLUSION: Rare clusters of inflammatory polyarthritis do occur. Their significance and cause remain to be elucidated. PMID- 10666176 TI - Influence of development and joint pathology on stromelysin enzyme activity in equine synovial fluid. AB - OBJECTIVE: To investigate the role of stromelysin (MMP-3) activity in synovial fluid (SF) at different stages of development and in common joint disorders in the horse. METHODS: Stromelysin activity was determined with a fluorogenic enzyme activity assay in SF of normal joints of fetal, juvenile and adult horses, and in SF of horses suffering from the developmental orthopaedic disease osteochondrosis (OC) or osteoarthritis (OA). Additionally, MMP-3 activity was expressed as a ratio of previously reported general MMP activity in the same SF samples. RESULTS: The levels of active stromelysin were 30-fold to 80-fold higher in SF from fetal horses than in SF from juvenile and mature animals (p<0.001). Juvenile horses (5 and 11 months of age) showed a twofold to threefold higher stromelysin activity than adult horses ( p<0.05). In OC joints, stromelysin activity was not significantly different from the activity in normal, age matched, control joints. In OA joints the activity was about four times higher than in normal joints (p<0.001). The ratio MMP-3 activity/general MMP activity did not change with age in normal, healthy joints. This ratio was more then twofold increased in OA joints compared with normal joints, indicating selective upregulation of gene expression or activation of proMMP-3, or both, in OA pathology. CONCLUSIONS: The significantly higher stromelysin activity in young individuals parallels the higher metabolic activity occurring at rapid growth and differentiation at early age. In OC, MMP-3 mediated matrix degradation appears to be not different from normal joints. The increased stromelysin activity in OA joints is in agreement with pathological matrix degradation. In these joints MMP-3 activity is selectively increased compared with normal joints. PMID- 10666177 TI - Relation of glenohumeral and acromioclavicular joint destruction in rheumatoid shoulder. A 15 year follow up study. AB - OBJECTIVES: To evaluate the relation of glenohumeral (GH) and acromioclavicular (AC) joint involvement in a cohort of 74 patients with seropositive and erosive rheumatoid arthritis (RA) followed up prospectively. METHODS: At the 15 year follow up radiographs of 148 shoulders were evaluated, and the grade of destruction of GH and AC joints were assessed by the Larsen method. One GH joint arthroplasty had been performed after 13 years of the disease onset and the preoperative radiograph was evaluated. RESULTS: Erosive involvement (Larsen grade >/= 2) was observed in 96 of 148 (65%) of the shoulders. Both GH and AC joints were affected in 62 of 148 (42%) shoulders. GH joint alone was involved in nine (6%) shoulders and only AC joint was affected in 25 (17%) shoulders. AC joint destruction correlated with the GH joint destruction, r=0.74 (95% confidence intervals (CI) 0.65 to 0.80 ). CONCLUSION: In RA AC joint is affected more often than the GH joint, but in half of the patients both joints are involved. This should be remembered when treating painful rheumatoid shoulder. PMID- 10666178 TI - Introduction: are natural anticoagulants candidates for modulating the inflammatory response to endotoxin? PMID- 10666179 TI - Recombinant human antithrombin III improves survival and attenuates inflammatory responses in baboons lethally challenged with Escherichia coli. AB - Plasma-derived antithrombin III (ATIII) prevents the lethal effects of Escherichia coli infusion in baboons, but the mechanisms behind this effect are not clear. In the present study, we evaluated the effects of recombinant human ATIII (rhATIII) on the clinical course and the inflammatory cytokine and coagulation responses in baboons challenged with lethal dose of E coli. Animals in the treatment group (n = 5) received high doses of rhATIII starting 1 hour before an E coli challenge. Those in the control group were administered saline. Survival was significantly improved in the treatment group (P =.002). Both groups had similar hemodynamic responses to E coli challenge but different coagulation and inflammatory responses. The rhATIII group had an accelerated increase of thrombin-ATIII complexes and significantly less fibrinogen consumption compared to controls. In addition, the rhATIII group had much less severe thrombotic pathology on autopsy and virtually no fibrinolytic response to E coli challenge. Furthermore, the rhATIII group had a significantly attenuated inflammatory response as evidenced by marked reduction of the release of various cytokines. We conclude that the early administration of high doses of rhATIII improves the outcome in baboons lethally challenged with E coli, probably due to the combined anticoagulation and anti-inflammatory effects of this therapy. (Blood. 2000;95:1117-1123) PMID- 10666180 TI - Tissue factor pathway inhibitor dose-dependently inhibits coagulation activation without influencing the fibrinolytic and cytokine response during human endotoxemia. AB - Inhibition of the tissue factor pathway has been shown to attenuate the activation of coagulation and to prevent death in a gram-negative bacteremia primate model of sepsis. It has been suggested that tissue factor influences inflammatory cascades other than the coagulation system. The authors sought to determine the effects of 2 different doses of recombinant tissue factor pathway inhibitor (TFPI) on endotoxin-induced coagulant, fibrinolytic, and cytokine responses in healthy humans. Two groups, each consisting of 8 healthy men, were studied in a double-blind, randomized, placebo-controlled crossover study. Subjects were studied on 2 different occasions. They received a bolus intravenous injection of 4 ng/kg endotoxin, which was followed by a 6-hour continuous infusion of TFPI or placebo. Eight subjects received 0.05 mg/kg per hour TFPI after a bolus of 0.0125 mg/kg (low-dose group), and 8 subjects received 0.2 mg/kg per hour after a bolus of 0.05 mg/kg (high-dose group). Endotoxin injection induced the activation of coagulation, the activation and subsequent inhibition of fibrinolysis, and the release of proinflammatory and antiinflammatory cytokines. TFPI infusion induced a dose-dependent attenuation of thrombin generation, as measured by plasma F1 + 2 and thrombin-antithrombin complexes, with a complete blockade of coagulation activation after high-dose TFPI. Endotoxin-induced changes in the fibrinolytic system and cytokine levels were not altered by either low-dose or high-dose TFPI. The authors concluded that TFPI effectively and dose-dependently attenuates the endotoxin-induced coagulation activation in humans without influencing the fibrinolytic and cytokine response. (Blood. 2000;95:1124-1129) PMID- 10666181 TI - Sickle cell anemia day hospital: an approach for the management of uncomplicated painful crises. AB - Painful crisis episodes are poorly treated in sickle cell anemia, both in timeliness and appropriateness of care. Delayed treatment in Emergency Departments, unrelieved pain, frequent admissions, and prolonged hospitalizations are common. We established a Day Hospital (DH) to determine if an alternative care delivery system could improve pain relief and reduce unnecessary hospital admissions for patients with uncomplicated painful crises. Trained DH staff delivered prompt titration for pain relief based on each patient's analgesic history and qualitative and quantitative assessments. Response to therapy and comorbidities commanded disposition. During the first 5 years of DH operation there were 2554 visits; 60% of the patients had severe pain. During an average visit of 4.5 hours, 84% of the patients were titrated to relief; 90% had pain relief within 2 to 4 hours. Overall, 81% of the patients were discharged home (70% initially and 90% to 94% in the last 3 years). During the first 5 years of the DH, there were 2612 emergency department (ED) visits that averaged 13 hours each. The combined ED and DH admissions during this time represented a 40% decrease in the baseline ED admission rate of 92%, (1 year pre-DH). Patients with uncomplicated painful crisis were admitted 5 times less often from the DH (8.3%) than from the ED (42.7%). The length of stay (LOS) for inpatients followed by the DH staff decreased by 1.5 days, while the LOS for patients followed by non-DH staff remained unchanged. Reduction of admissions and LOS represented a savings of approximately $1.7 million. We conclude that a dedicated facility provides the kingpin for effective and rapid painful crisis management, reduces hospitalizations, and facilitates integration of the approach into other areas of care. (Blood. 2000;95:1130-1136) PMID- 10666182 TI - Simple PCR detection of haptoglobin gene deletion in anhaptoglobinemic patients with antihaptoglobin antibody that causes anaphylactic transfusion reactions. AB - Two anhaptoglobinemic patients showing anaphylactic transfusion reactions by antihaptoglobin antibody were found. Southern blot analysis indicated that 2 patients were homozygous for the deleted allele of the haptoglobin gene (Hp(del)) as reported previously. We have identified the junction region of the deletion from genomic DNA of 1 patient using cassette-mediated polymerase chain reaction (PCR). Then, the deleted region from the 5' breakpoint to the promoter region of the Hp was amplified from genomic DNA of a control individual using PCR. DNA sequence analysis of these regions indicated that the 5' breakpoint of the Hp(del) allele was located 5. 2 kilobase (kb) upstream of exon 1 of the Hp and the 3' breakpoint was positioned between 52 and 53 base pair (bp) upstream of exon 5 of the haptoglobin-related gene. There was no significant homology between the DNA sequences flanking the 5' and 3' breakpoints, except for a 2-bp (TG) identity. To examine the gene frequency, we have developed a simple PCR method to detect the gene deletion. We found 8, 16, and 17 Hp(del) alleles in 157 Koreans, 523 Japanese, and in 284 Chinese, respectively, but did not find the Hp(del) in 101 Africans or in 100 European-Africans. The incidence of individuals homozygous for the Hp(del) allele was therefore expected to be 1/4000 in Japanese, 1/1500 in Koreans, and 1/1000 in Chinese. This incidence is higher than that of IgA deficiency in Japanese. More attention should be paid on haptoglobin deficiency and antihaptoglobin antibody as the cause of transfusion-related anaphylactic reactions in Asian populations. (Blood. 2000;95:1138-1143) PMID- 10666183 TI - Acquired loss of p53 induces blastic transformation in p210(bcr/abl)-expressing hematopoietic cells: a transgenic study for blast crisis of human CML. AB - Chronic myelogenous leukemia (CML) begins with an indolent chronic phase but inevitably progresses to a fatal blast crisis. Although the Philadelphia chromosome, which generates p210(bcr/abl), is a unique chromosomal abnormality in the chronic phase, additional chromosomal abnormalities are frequently detected in the blast crisis, suggesting that superimposed genetic events are responsible for disease progression. To investigate whether loss of p53 plays a role in the evolution of CML, we crossmated p210(bcr/abl)-transgenic (BCR/ABL(tg/-)) mice with p53-heterozygous (p53(+/-)) mice and generated p210(bcr/abl)-transgenic, p53 heterozygous (BCR/ABL(tg/-)p53(+/-)) mice, in which a somatic alteration in the residual normal p53 allele directly abrogates p53 function. The BCR/ABL(tg/ )p53(+/-) mice died in a short period compared with their wild-type (BCR/ABL(-/ )p53(+/+)), p53 heterozygous (BCR/ABL(-/-)p53(+/-)), and p210(bcr/abl) transgenic (BCR/ABL(tg/-)p53(+/+)) litter mates. They had rapid proliferation of blast cells, which was preceded by subclinical or clinical signs of a myeloproliferative disorder resembling human CML. The blast cells were clonal in origin and expressed p210(bcr/abl) with an increased kinase activity. Interestingly, the residual normal p53 allele was frequently and preferentially lost in the tumor tissues, implying that a certain mechanism facilitating the loss of p53 allele exists in p210(bcr/abl)-expressing hematopoietic cells. Our study presents in vivo evidence that acquired loss of p53 contributes to the blastic transformation of p210(bcr/abl)-expressing hematopoietic cells and provides insights into the molecular mechanism for blast crisis of human CML. (Blood. 2000;95:1144-1150) PMID- 10666184 TI - KSHV-encoded CC chemokine vMIP-III is a CCR4 agonist, stimulates angiogenesis, and selectively chemoattracts TH2 cells. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) encodes 3 genes that are homologous to cellular chemokines. vMIP-III, the product of open reading frame K4.1, is the most distantly related to human chemokines and has yet to be characterized. We have examined the interaction of vMIP-III with chemokine receptors, its expression in KS lesions, and its in ovo angiogenic properties. We show expression of vMIP-III in KS lesions and demonstrate the stimulation of angiogenesis by this chemokine, like vMIP-I and vMIP-II, in the chick chorioallantoic membrane assay. vMIP-III does not block human immunodeficiency virus entry through the coreceptors CCR3, CCR5, or CXCR4. However, vMIP-III is an agonist for the cellular chemokine receptor CCR4. CCR4 is expressed by TH2-type T cells. Consistent with this, vMIP-III preferentially chemoattracts this cell type. Because of these biologic properties and because it is expressed in KS lesions, vMIP-III may play an important role in the pathobiology of KS. (Blood. 2000;95:1151-1157) PMID- 10666185 TI - The CXC-chemokine platelet factor 4 promotes monocyte survival and induces monocyte differentiation into macrophages. AB - Unstimulated monocytes rapidly undergo physiological changes resulting in programmed cell death (apoptosis) while stimuli promoting differentiation of these cells into macrophages were shown to inhibit apoptotic processes. In the present study, we report that the platelet-derived alpha-chemokine platelet factor 4 (PF4) induces the differentiation of monocytes into macrophages, as is evident from morphological changes as well as from the up-regulation of differentiation markers (carboxypeptidase M/MAX1 and CD71). Significant alterations of the phenotype were observed after 72 hours of culture in the presence of the chemokine and required a minimal concentration of 625 nmol/L PF4. PF4-induced macrophages were characterized by a lack of HLA-DR antigen on their surface but showed a strong increase in the expression of the CD28 ligand B7-2. Furthermore, PF4 stimulation prevented monocytes from undergoing spontaneous apoptosis during 72 hours of culture as determined in an annexin-V-binding assay. Although PF4 induced the secretion of relevant amounts of TNF-alpha, neutralizing antibodies directed against TNF-alpha or granulocyte-macrophage colony stimulating factor (GM-CSF) did not revert PF4-induced rescue from programmed cell death, suggesting that PF4 exerts its antiapoptotic effects in a TNF-alpha- or GM-CSF-independent fashion. On the basis of these results, we propose a novel role for PF4 in the control of monocyte differentiation during an inflammatory process in vivo. (Blood. 2000;95:1158-1166) PMID- 10666186 TI - Limited expression of R5-tropic HIV-1 in CCR5-positive type 1-polarized T cells explained by their ability to produce RANTES, MIP-1alpha, and MIP-1beta. AB - Human T helper (Th) cells (Th1- or Th2-oriented memory T cells as well as Th1- or Th2-polarized naive T cells) were infected in vitro with an R5-tropic HIV-1 strain (BaL) and assessed for their profile of cytokine production, CCR5 receptor expression, and HIV-1 p24 antigen (p24 Ag) production. Higher p24 Ag production was found in CCR5-negative Th2-like memory T cells than in CCR5-positive Th1-like memory T cells. By contrast, p24 Ag production was higher in Th1-polarized activated naive T cells in the first 4 days after infection. However, p24 Ag production in Th1-polarized T cells became comparable or even lower than the production in Th2-polarized populations later in infection or when the cells were infected with HIV-1BaL after secondary stimulation. The higher levels of p24 Ag production by Th1-polarized naive T cells soon after infection reflected a higher virus entry, as assessed by the single round infection assay using the HIV chloramphenicol acetyl transferase (HIV-CAT) R5-tropic virus that contains the envelope protein of HIV-1 YU2 strain. The limitation of viral spread in the Th1 polarized populations, despite the initial higher level of T-cell entry of R5 tropic strains, was due to the ability of Th1 cells to produce greater amounts of beta-chemokines than Th2 cells. In fact, an inverse correlation was observed between Th1-polarized naive T cells and Th1-like memory-activated T cells in regards to p24 Ag production and the release of the following CCR5-binding chemokines: regulated on activation normal T expressed and secreted (RANTES), macrophage inflammatory protein-1alpha (MIP-1alpha), and MIP-1beta. Moreover, infection with the HIV-1BaL strain of Th1-polarized T cells in the presence of a mixture of anti-RANTES, anti-MIP-1alpha, and anti-MIP-1beta neutralizing antibodies resulted in a significant increase of HIV-1 expression. These findings suggest that Th1-type responses may favor CD4(+) T-cell infection by R5-tropic HIV-1 strains, but HIV-1 spread in Th1 cells is limited by their ability to produce CCR5-binding chemokines. (Blood. 2000;95:1167-1174) PMID- 10666187 TI - Effect of recombinant human erythropoietin combined with granulocyte/ macrophage colony-stimulating factor in the treatment of patients with myelodysplastic syndrome. GM/EPO MDS Study Group. AB - This randomized, placebo-controlled trial was designed to assess the efficacy and safety of therapy with granulocyte-macrophage colony-stimulating factor (GM-CSF) and erythropoietin (epoetin alfa) in anemic, neutropenic patients with myelodysplastic syndrome. Sixty-six patients were enrolled according to the following French-American-British classification: refractory anemia (20), refractory anemia with excess blasts (35), refractory anemia with ringed sideroblasts (9), and refractory anemia with excess blasts in transformation (2). Patients were stratified by their serum erythropoietin levels (less than or equal to 500 mU/mL, n = 37; greater than 500 mU/mL, n = 29) and randomized, in a 2:1 ratio, to either GM-CSF (0.3-5.0 microg/kg.d) + epoetin alfa (150 IU/kg 3 times/wk) or GM-CSF (0.3-5.0 microg/kg.d) + placebo (3 times/wk). The mean neutrophil count rose from 948 to 3831 during treatment with GM-CSF +/- epoetin alfa. Hemoglobin response (increase greater than or equal to 2 g/dL, unrelated to transfusion) occurred in 4 of 45 (9%) patients in the GM-CSF + epoetin alfa group compared with 1 of 21 (5%) patients with GM-CSF + placebo group (P = NS). Percentages of patients in the epoetin alfa and the placebo groups requiring transfusions of red blood cells were 60% and 92%, respectively, for the low endogenous erythropoietin patients and 95% and 89% for the high-endogenous erythropoietin patients (P = NS). Similarly, the average numbers of units of red blood cells transfused during the 12-week study in the epoetin alfa and the placebo groups were 5.9 and 9.5, respectively, in the low-endogenous erythropoietin patients and 9.7 and 8.6 in the high-endogenous erythropoietin patients (P = NS). GM-CSF +/- epoetin alfa had no effect on mean platelet count. Treatment was well tolerated in most patients, though 10 withdrew from the study for reasons related predominantly to GM-CSF toxicity. (Blood. 2000;95:1175-1179) PMID- 10666188 TI - Genetic aberrations common in gastric high-grade large B-cell lymphoma. AB - Genetic aberrations associated with the development of extranodal high-grade large B-cell lymphoma originating in the stomach have not been fully identified yet. We analyzed 31 such lymphomas using 73 microsatellite markers for allelic imbalance and microsatellite instability. The highest frequency (42%) of loss of heterozygosity (LOH) was found on the long arm of chromosome 6. We identified 2 LOH hot spots on 6q21-22.1 and 6q23.3-25, flanked by markers D6S246-D6S261 and D6S310-D6S441, respectively, containing putative tumor suppressor genes (TSGs). These 6q aberrations were found to be the sole allelic imbalance in 1 patient only; they were mostly accompanied by additional abnormalities. Several known TSGs, namely, the APC, p15/p16, p53, and DCC genes, were found to suffer frequent LOH during lymphomagenesis. LOH was also detected in regions containing putative TSGs on 7q and 13q14. Frequent amplification of genomic material was found in the 2p, 3q27 at the BCL-6 gene locus, 6p, 7q, 11q23-24 at the MLL gene locus, and 18q regions. Analysis of the pattern of occurrence of these aberrations revealed an association of the amplification of the MLL gene region with LOH at the p53 locus (P =.02). Only low frequency of microsatellite instability (MSI) was detected in these lymphomas and MSI incidence increased with age (P =.01). Karyotypic instability thus plays the main role in the development of gastric high-grade large B-cell lymphoma. Common genetic aberrations responsible for lymphomagenesis are deletions of 6q, loss of p53, and amplification of the 3q27 and the MLL gene regions. (Blood. 2000;95:1180-1187) PMID- 10666189 TI - Allogeneic and syngeneic marrow transplantation for myelodysplastic syndrome in patients 55 to 66 years of age. AB - We carried out bone marrow transplantation (BMT) in 50 patients with myelodysplastic syndrome (MDS) who were 55.3 to 66.2 years of age (median, 58.8 years). According to the criteria of the French-American-British (FAB) classification, 13 patients had refractory anemia (RA), 19 had RA with excess blasts (RAEB), 16 had RAEB in transformation or acute myelogenous leukemia (RAEB T/AML), and 2 had chronic myelomonocytic leukemia (CMML). According to the recently established International Prognostic Scoring System (IPSS), available for 45 patients, 2 patients were considered low risk; 14, intermediate 1 risk; 19, intermediate 2 risk; and 10, high risk. Conditioning regimens were cyclophosphamide (CY) (120 mg/kg of body weight) plus 12-Gy fractionated total body irradiation (FTBI) (n = 15), CY plus FTBI with lung and liver shielding (n = 4), busulfan (7 mg/kg) plus FTBI (n = 4), or busulfan (16 mg/kg) plus CY (n = 27). The busulfan-plus-CY group included 16 patients in whom busulfan was targeted to plasma levels of 600 to 900 ng/mL. In these 16 patients, steady-state levels of busulfan actually achieved were 714 to 961 ng/mL (mean +/- SD, 845 +/- 64 ng/mL; median, 838 ng/mL). The donors were HLA-identical siblings for 34 patients, HLA-nonidentical family members for 4, identical twins for 4, and unrelated volunteers for 6. All 46 patients surviving > 21 days had engraftment, and 22 patients (44%) are surviving 9 to 80 months after BMT. Specifically, among 13 patients with RA, 1 had relapse (cumulative incidence [CI] at 3 years, 8%) and 8 are surviving, for a Kaplan-Meier (KM) estimate of survival at 3 years of 59% (disease-free survival [DSF], 53%). Among 19 patients with RAEB, 3 had relapse (CI at 3 years, 16%), and 8 are surviving disease free (KM estimate at 3 years, 46%). Among 18 patients with RAEB-T/AML or CMML, 6 had relapse (CI at 3 years, 28%), and the KM estimate of DSF at 3 years is 33%. Relapse-free survival had an inverse correlation with cytogenetic risk classification and with the risk score according to the IPSS. Survival in all FAB categories was highest among patients enrolled in a protocol in which busulfan plasma levels were targeted to 600 to 900 ng/mL. These data indicate that BMT can be carried out successfully in patients with MDS who are older than 55 years of age. (Blood. 2000;95:1188-1194) PMID- 10666190 TI - Stem cell transplantation in patients with severe congenital neutropenia without evidence of leukemic transformation. AB - Severe congenital neutropenia (CN) (Kostmann syndrome) is a hematologic disorder characterized by a maturation arrest of myelopoiesis at the promyelocyte/myelocyte stage of development. This arrest results in severe neutropenia leading to absolute neutrophil counts (ANC) below 0.2 x 10(9)/L associated with severe bacterial infections from early infancy. Data on over 300 patients with CN collected by the Severe Chronic Neutropenia International Registry (SCNIR) beginning in 1994 indicate that more than 90% of these patients respond to recombinant human granulocyte-colony stimulating factor (r-HuG-CSF) treatment with an ANC greater than 1. 0 x 10(9)/L. For patients who are refractory to r-HuG-CSF treatment and continue to have severe and often life threatening bacterial infections, hematopoietic stem cell transplantation is the only currently available treatment. We report on a total of 11 patients with CN reported to the SCNIR who underwent transplantation for reasons other than malignant transformation between 1976 and 1998. Of these patients, 8 were nonresponders or showed only partial response to r-HuG-CSF treatment with ongoing infections. Results from these patients suggest that transplantation of stem cells from an HLA-identical sibling is beneficial for patients refractory to r HuG-CSF. (Blood. 2000;95:1195-1198) PMID- 10666191 TI - Apoptosis or plasma cell differentiation of CD38-positive B-chronic lymphocytic leukemia cells induced by cross-linking of surface IgM or IgD. AB - Previously, we demonstrated that B-chronic lymphocytic leukemia (B-CLL) cells could be divided into 2 groups depending on the expression of CD38 by the malignant cells. The 2 groups differed in their signal-transducing capacities initiated by cross-linking of surface IgM; only in CD38-positive cells was an efficient signal delivered, invariably resulting in cell apoptosis. In this study, we investigated the effect of surface IgD cross-linking in 10 patients with CD38-positive B-CLL. Exposure of the malignant cells to goat antihuman delta chain antibodies (Gadelta-ab) caused [Ca(++)]i mobilization and tyrosine kinase phosphorylation in a manner not different from that observed after goat antihuman mu-chain antibody (Gamu-ab) treatment in vitro. However, Gadelta-ab-treated cells failed to undergo apoptosis and instead displayed prolonged survival in culture and differentiated into plasma cells when rIL2 was concomitantly present. Cross linking of surface IgD failed to induce proliferation of the malignant cells in vitro. Moreover, treatment with Gadelta-ab did not prevent apoptosis of B-CLL cells induced by Gamu-ab. Collectively, these experiments demonstrated that IgM and IgD expressed by the same cell may deliver opposite signals under particular circumstances and provide some clues for the understanding of the pathophysiology of B-CLL. (Blood. 2000;95:1199-1206) PMID- 10666192 TI - Tissue eosinophilia correlates strongly with poor prognosis in nodular sclerosing Hodgkin's disease, allowing for known prognostic factors. AB - Although eosinophilic granulocytes are frequently observed in lymphatic tissue of Hodgkin's patients, no substantial data reveal the prognostic role, if any, of tissue eosinophilia. Thus, eosinophilia was analyzed histologically in 1511 diagnostic biopsy specimens of patients treated under protocol therapy of the German Hodgkin's Lymphoma Study Group between 1988 and 1994. Prominent eosinophilia was seen in 38% of cases, which differed among the histologic types of Hodgkin's disease (HD): none in lymphocyte predominant, 14% in lymphocyte rich classical, 40% in nodular sclerosis grade 1 (NS-1), 55% in nodular sclerosis grade 2, 43% in mixed cellularity (MC), and 54% in lymphocyte depleted. In a multivariate analysis, tissue eosinophilia proved to be the strongest prognostic factor for freedom from treatment failure (P <. 001) and overall survival (P <.001) in a stage-stratified model. Among NS-1 patients, the effect was highly significant. In MC, no significant effect of eosinophilia on survival could be demonstrated. Eosinophils secrete CD30 ligand that is capable of binding to CD30 positive HD cells. The activation of TRAF2, followed by NF-kappaB, which occurs on CD30L/CD30 binding, may explain the neoplastic proliferation and apoptosis protection of HD cells. TRAF2 is also activated by EBV-LMP expression, which is detectable in the majority of MC but not NS cases. In addition to the possibility that eosinophils are only passive indicators for other unknown prognostic determinants, it may be concluded that the positive clinical outcome of eosinophilia-negative NS cases could be due to lower NF-kappaB activity. (Blood. 2000;95:1207-1213) PMID- 10666193 TI - Treatment of relapsed leukemia after unrelated donor marrow transplantation with unrelated donor leukocyte infusions. AB - The efficacy and toxicity of donor leukocyte infusions (DLI) after unrelated donor bone marrow transplantation (BMT) is largely unknown. We identified 58 recipients of unrelated DLI (UDLI) for the treatment of relapsed disease from the National Marrow Donor Program database. A retrospective analysis was performed to determine response, toxicity, and survival after UDLI and to identify factors associated with successful therapy. UDLI was administered for relapsed chronic myelogenous leukemia (CML) (n = 25), acute myelogenous leukemia (AML) (n = 23), acute lymphoblastic leukemia (ALL) (n = 7), and other diseases (n = 3). Eight patients were in complete remission (CR) before UDLI, and 50 were evaluable for response. Forty-two percent (95% confidence interval [CI], 28%-56%) achieved CR, including 11 of 24 (46%; 95% CI, 26%-66%) with CML, 8 of 19 (42%; 95% CI, 20% 64%) with AML, and 2 of 4 (50%; 95% CI, 1%-99%) with ALL. The estimated probability of disease-free survival (DFS) at 1 year after CR was 65% (95% CI, 50%-79%) for CML, 23% (95% CI, 9%-38%) for AML, and 30% (95% CI, 6%-54%) for ALL. Acute graft-versus-host disease (GVHD) complicated UDLI in 37% of patients (grade II-IV, 25%). A total of 13 of 32 evaluable patients (41%) developed chronic GVHD. There was no association between cell dose administered and either response or toxicity. In a multivariable analysis, only a longer interval from BMT to relapse and BMT to UDLI was associated with improved survival and DFS, respectively. UDLI is an acceptable alternative to other treatment options for relapse after unrelated donor BMT. (Blood. 2000;95:1214-1221) PMID- 10666194 TI - Polymorphisms within glutathione S-transferase genes (GSTM1, GSTT1, GSTP1) and risk of relapse in childhood B-cell precursor acute lymphoblastic leukemia: a case-control study. AB - Glutathione S-transferases (GSTs) have been associated with outcome in human cancers treated with cytotoxic chemotherapy. In a case-control study, we investigated the association between polymorphisms within the GSTM1, GSTT1, and GSTP1 genes and risk of relapse in childhood acute lymphoblastic leukemia (ALL). Cases were relapsed patients. Controls were successfully treated patients with a minimum follow-up of 5 years. The null genotype (absence of both alleles) for GSTM1 or GSTT1 conferred a 2-fold (OR = 0.5, 95% CI = 0. 23-1.07, P =.078) and 2.8-fold (OR = 0.36, 95% CI = 0.13-0.99, P =. 048) reduction in risk of relapse, respectively, relative to the presence of the GSTM1 or GSTT1 gene. The GSTP1 Val(105)/Val(105) genotype showed a 3-fold decrease in risk of relapse (OR = 0.33, 95% CI = 0.09-1.23, P =.099) in comparison to the combined category of Ile(105)/Val(105) and Ile(105)/Ile(105 )genotypes. No particular associations with relapse were observed for the GSTP1 polymorphism at codon 114. The risk of relapse when having 1 of the low-risk genotypes (GSTM1 null, GSTT1 null, GSTP1 Val(105)/Val(105)) decreased 1.9-fold (OR = 0.53, 95% CI = 0.24-1.19, P =.123), and the risk when having 2 or 3 low-risk genotypes 3.5-fold (OR = 0.29, 95% CI = 0.06-1.37, P =.118), compared with individuals having no low-risk genotype (P for trend =.005). Our results suggest that polymorphisms within genes of the GST superfamily may be associated with risk of relapse in childhood ALL. (Blood. 2000;95:1222-1228) PMID- 10666195 TI - Long-term outcome of continuous 24-hour deferoxamine infusion via indwelling intravenous catheters in high-risk beta-thalassemia. AB - The optimal regimen of intravenous deferoxamine for iron overload in high-risk homozygous beta-thalassemia is unknown because only short-term follow-up has been described in small patient groups. We report the outcome over a 16-year period of a continuous 24-hour deferoxamine regimen, with dose adjustment for serum ferritin, delivered via 25 indwelling intravenous lines for 17 patients. Treatment indications were cardiac arrhythmias, left ventricular dysfunction, gross iron overload, and intolerability of subcutaneous deferoxamine. Cardiac arrhythmias were reversed in 6 of 6 patients, and the left ventricular ejection fraction improved in 7 of 9 patients from a mean (+/- SEM) of 36 +/- 2% to 49 +/- 3% (P =.002, n = 9). The serum ferritin fell in a biphasic manner from a pretherapy mean of 6281 +/- 562 microg/L to 3736 +/- 466 microg/L (P =.001), falling rapidly and proportionally to the pretreatment ferritin (r(2) = 0.99) for values >3000 microg/L but falling less rapidly below this value (at 133 +/- 22 microg/L/mo). The principal catheter-related complications were infection and thromboembolism (1. 15 and 0.48 per 1000 catheter days, respectively), rates similar to other patient groups. Only one case of reversible deferoxamine toxicity was observed (retinal) when the therapeutic index was briefly exceeded. An actuarial survival of 61% at 13 years with no treatment-related mortality provides evidence of the value of this protocol. (Blood. 2000;95:1229-1236) PMID- 10666196 TI - Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. AB - Mobilized peripheral blood progenitor cells (PBPC) are a potential target for the retrovirus-mediated transfer of cytostatic drug-resistance genes. We analyzed nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse-repopulating CD34+ PBPC from patients with cancer after retroviral transduction in various cytokine combinations with the hybrid vector SF-MDR, which is based on the Friend mink cell focus-forming/murine embryonic stem-cell virus and carries the human multidrug resistance 1 (MDR1) gene. Five to 13 weeks after transplantation of CD34+ PBPC into NOD/SCID mice (n = 84), a cell dose-dependent multilineage engraftment of human leukocytes up to an average of 33% was observed. The SF-MDR provirus was detected in the bone marrow (BM) and in its granulocyte fractions in 96% and 72%, respectively, of chimeric NOD/SCID mice. SF-MDR provirus integration assessed by quantitative real-time polymerase chain reaction (PCR) was optimal in the presence of Flt-3 ligand/thrombopoietin/stem-cell factor, resulting in a 6 fold (24% +/- 5% [mean +/- SE]) higher average proportion of gene-marked human cells in NOD/SCID mice than that achieved with IL-3 alone (P <.01). A population of clearly rhodamine-123(dull) human myeloid progeny cells could be isolated from BM samples from chimeric NOD/SCID mice. On the basis of PCR and rhodamine-123 efflux data, up to 18% +/- 4% of transduced cells were calculated to express the transgene. Our data suggest that the NOD/SCID model provides a valid assay for estimating the gene-transfer efficiency to repopulating human PBPC that may be achievable in clinical autologous transplantation. P-glycoprotein expression sufficient to prevent marrow aplasia in vivo may be obtained with this SF-MDR vector and an optimized transduction protocol. (Blood. 2000;95:1237-1248) PMID- 10666197 TI - Targeted disruption of stat6 DNA binding activity by an oligonucleotide decoy blocks IL-4-driven T(H)2 cell response. AB - The transcription factor, signal transducer and activator of transcription (Stat) 6, regulates T(H)2-lymphocyte activity by controlling the expression and responsiveness to interleukin (IL)-4, which plays a key role in numerous allergic maladies. Therefore, we sought to use a phosphorothiolate cis-element decoy to target disruption of Stat6 transcriptional activity. Here we showed that the Stat6 decoy potently ablated the messenger RNA expression and production of IL-4, but not of several other cytokines. The Stat6 decoy functionally disrupted IL-4 inducible cell proliferation of murine T(H)2 cells and primary human CD4(+) T lymphocytes. Specificity of the decoy was demonstrated by its ability to directly block Stat6 binding to a cis-element probe and transactivation, but not affect Stat6 tyrosine phosphorylation or expression of the IL-4 receptor chains. Moreover, the decoy failed to inhibit non-Stat6-dependent signaling pathways since IL-2 was competent to induce cell proliferation and activation of Stats 1, 3, and 5a/b. With the use of laser scanning confocal microscopy, fluorescently tagged Stat6 decoy was detectable in the cytoplasm and nucleus; however, greater levels of oligonucleotide were present in the latter following IL-4 treatment. Taken together, these data suggest that IL-4-driven T(H)2 cell activity can be preferentially restricted via targeted disruption of Stat6 by a novel and specific decoy strategy that may possess gene therapeutic potential. (Blood. 2000;95:1249-1257) PMID- 10666198 TI - An oral CD40 ligand gene therapy against lymphoma using attenuated Salmonella typhimurium. AB - CD40 ligand (CD40L) has a great potential as a novel treatment for B-cell lymphoma (BCL). It has previously been demonstrated that a nonvirulent strain of Salmonella typhimurium mutant (ST) can be used not only as a vehicle in oral genetic immunization via the intestinal mucosa, but also as an enhancer of interferon gamma- and tumor necrosis factor alpha-mediated immunity. After confirming that human CD40L can up-regulate expression of Fas, B7-1, and B7-2 molecules on murine BCL cells in vitro, we transfected the human CD40L gene into S typhimurium mutant (ST40L), which was administrated orally to determine whether it was able to prevent the growth of BCL in mice. Expression of human CD40L was confirmed immunohistochemically with protein being detected in the Peyer's patches of mice immunized with ST40L. Moreover, human soluble CD40L had been detectable until 7 to 8 weeks after oral administration of ST40L. Although ST alone exhibited some protective effects, ST40L demonstrated a significantly greater protection against the development of CD40 positive BCL compared with the control. In the surviving mice that had been treated with ST40L, a small and hard nodule was formed at the injection site, which was found to be composed of infiltrating lymphocytes expressing Fas ligand. These results have the potential to be a simple, effective, and above all, safe immune-gene therapy against BCL. (Blood. 2000;95:1258-1263) PMID- 10666199 TI - GATA-1 blocks IL-6-induced macrophage differentiation and apoptosis through the sustained expression of cyclin D1 and bcl-2 in a murine myeloid cell line M1. AB - Cytokines exert pleiotropic effects on target cells in a manner dependent on the cell type or stage of differentiation. To determine how instinctive cell properties affect biological effects of cytokine, we introduced an erythroid/megakaryocyte lineage-specific transcription factor, GATA-1, into a murine myeloid cell line M1, which is known to undergo macrophage differentiation in response to interleukin 6 (IL-6). Overexpression of GATA-1 changed the phenotype of M1 cells from myeloid to megakaryocytic lineage. Furthermore, GATA-1 blocked both IL-6-induced macrophage differentiation and apoptosis of M1 cells. Although STAT3 is essential for IL-6-induced macrophage differentiation of M1 cells, GATA-1 had little or no effect on tyrosine phosphorylation, DNA binding, and transcriptional activities of STAT3 in Western blot analysis, electropholic mobility shift assay (EMSA), and luciferase assays. During IL-6-induced macrophage differentiation of M1 cells, IL-6 down-regulated cyclin D1 expression and induced p19(INK4D) expression, leading to reduction in cdk4 activities. In contrast, sustained expression of cyclin D1 and a significantly lesser amount of p19(INK4D) induction were observed in IL-6-treated M1 cells overexpressing GATA 1. Furthermore, although bcl-2 expression was severely reduced by IL-6 in M1 cells, it was sustained in GATA-1-introduced M1 cells during the culture with IL 6. Both IL-6-induced macrophage differentiation and apoptosis were significantly abrogated by coexpression of cyclin D1 and bcl-2, whereas overexpressions of cyclin D1 or bcl-2 inhibited only differentiation or apoptosis, respectively. These results suggested that GATA-1 may not only reprogram the lineage phenotype of M1 cells but also disrupt the biologic effects of IL-6 through the sustained expression of cyclin D1 and bcl-2. (Blood. 2000;95:1264-1273) PMID- 10666200 TI - Hemopoietic lineage commitment decisions: in vivo evidence from a transgenic mouse model harboring micro LCR-betapro-LacZ as a transgene. AB - A substantial body of published data suggests activation of lineage-specific genes in multipotential hemopoietic cells before their unilineage commitment. Because the behavior and plasticity of cells isolated in vitro away from microenvironmental constraints exercised in vivo may be altered, one wonders whether similar findings can be observed in a physiologic setting in vivo. We used a transgenic mouse model harboring human micro LCR together with beta promoter sequences as a transgene to examine activation of lineage-specific programs in vivo. By using LacZ as a reporter, we had the ability to detect, quantitate, and select live cells with different levels of LacZ activation. We found strong expression of LacZ by X-gal staining in 2 lineages-erythroid and megakaryocytic. Activation in the latter was a novel finding not previously observed when similar transgenes were used. We also found activation of muLCR betapro at low levels in progenitor cells of granulocytic-macrophagic, erythroid, or megakaryocytic lineage detected by in vitro assays, suggesting activation before commitment to a specific lineage pathway. In particular, the expression of LacZ was graded among progenitors, so that in a proportion of them activation occurred only after commitment to erythroid or megakaryocytic lineage. In addition, we found quantitative reduction in LacZ expression between fetal liver and bone marrow-derived cells, the basis of which is unclear. Collectively our data provide in vivo evidence supporting the view that lineage-specific genes are expressed in a graded fashion in pluripotential cells before their irreversible unilineage commitment. (Blood. 2000;95:1274-1282) PMID- 10666201 TI - Actin cytoskeletal function is spared, but apoptosis is increased, in WAS patient hematopoietic cells. AB - Mutations in the Wiskott-Aldrich syndrome protein (WASP) have been hypothesized to cause defective actin cytoskeletal function. This resultant dysfunction of the actin cytoskeleton has been implicated in the pathogenesis of Wiskott-Aldrich syndrome (WAS). In contrast, it was found that stimulated actin polymerization is kinetically normal in the hematopoietic lineages affected in WAS. It was also found that the actin cytoskeleton in WAS platelets is capable of producing the hallmark cytoarchitectural features associated with activation. Further analysis revealed accelerated cell death in WAS lymphocytes as evidenced by increased caspase-3 activity. This increased activity resulted in accelerated apoptosis of these cells. CD95 expression was also increased in these cells, suggesting an up regulation in the FAS pathway in WAS lymphocytes. Additionally, inhibition of actin polymerization in lymphocytes using cytochalasin B did not accelerate apoptosis in these cells. This suggests that the accelerated apoptosis observed in WAS lymphocytes was not secondary to an underlying defect in actin polymerization caused by mutation of the WAS gene. These data indicate that WASP does not play a universal role in signaling actin polymerization, but does play a role in delaying cell death. Therefore, the principal consequence of mutations in the WAS gene is to accelerate lymphocyte apoptosis, potentially through up regulation of the FAS-mediated cell death pathway. This accelerated apoptosis may ultimately give rise to the clinical manifestations observed in WAS. (Blood. 2000;95:1283-1292) PMID- 10666202 TI - Adherence of phosphatidylserine-exposing erythrocytes to endothelial matrix thrombospondin. AB - Phospholipid asymmetry is well maintained in erythrocyte (RBC) membranes with phosphatidylserine (PS) exclusively present in the inner leaflet. The appearance of PS on the surface of the cell can have major physiologic consequences, including increased cell-cell interactions. Because increased adherence of PS exposing RBCs to endothelial cells (ECs) may be pathologically important in hemoglobinopathies such as sickle cell disease and thalassemia, we studied the role of PS exposure in calcium ionophore-treated normal RBC adherence to human umbilical vein endothelial cell (HUVEC) monolayers. When HUVEC monolayers were incubated with these PS-exposing RBCs, the ECs retracted and the RBCs adhered primarily in the gaps opened between the ECs. A linear correlation was found between the number of PS-exposing RBCs in the population and the number of adhering RBCs to the monolayer. Pretreatment of RBCs with annexin V significantly decreased adherence by shielding PS on the RBCs. Similarly, PS-containing lipid vesicles decreased RBC binding by competing for the PS binding sites in the monolayer. PS-exposing RBCs and PS-containing lipid vesicles adhered to immobilized thrombospondin (TSP) and matrix TSP, respectively, and adherence of PS-exposing RBCs to EC monolayers was reduced by antibodies to TSP and to its EC receptor, alpha(v)beta(3). Together, these results indicate a role for PS and matrix TSP in the adherence of PS-exposing RBCs to EC monolayers, and suggest an important contribution of PS-exposing RBCs in pathologies with reported vascular damage, such as sickle cell anemia. (Blood. 2000;95:1293-1300) PMID- 10666203 TI - Streptokinase-induced platelet activation involves antistreptokinase antibodies and cleavage of protease-activated receptor-1. AB - Streptokinase activates platelets, limiting its effectiveness as a thrombolytic agent. The role of antistreptokinase antibodies and proteases in streptokinase induced platelet activation was investigated. Streptokinase induced localization of human IgG to the platelet surface, platelet aggregation, and thromboxane A(2) production. These effects were inhibited by a monoclonal antibody to the platelet Fc receptor, IV.3. The platelet response to streptokinase was also blocked by an antibody directed against the cleavage site of the platelet thrombin receptor, protease-activated receptor-1 (PAR-1), but not by hirudin or an active site thrombin inhibitor, Ro46-6240. In plasma depleted of plasminogen, exogenous wild type plasminogen, but not an inactive mutant protein, S(741)A plasminogen, supported platelet aggregation, suggesting that the protease cleaving PAR-1 was streptokinase-plasminogen. Streptokinase-plasminogen cleaved a synthetic peptide corresponding to PAR-1, resulting in generation of PAR-1 tethered ligand sequence and selectively reduced binding of a cleavage-sensitive PAR-1 antibody in intact cells. A combination of streptokinase, plasminogen, and antistreptokinase antibodies activated human erythroleukemic cells and was inhibited by pretreatment with IV.3 or pretreating the cells with the PAR-1 agonist SFLLRN, suggesting Fc receptor and PAR-1 interactions are necessary for cell activation in this system also. Streptokinase-induced platelet activation is dependent on both antistreptokinase-Fc receptor interactions and cleavage of PAR-1. (Blood. 2000;95:1301-1308) PMID- 10666204 TI - Transcriptional activation of urokinase by the Kruppel-like factor Zf9/COPEB activates latent TGF-beta1 in vascular endothelial cells. AB - Understanding the regulation of genes controlling fibrinolysis and matrix homeostasis is essential for elucidating the basis of tissue repair. A recently described novel Kruppel-like factor, Zf9, is up-regulated in acute liver injury in activated hepatic stellate cells. Because Zf9 can be induced widely, its activity was examined in vascular endothelium, a key cell in vascular injury. Zf9 is induced as an immediate-early response gene in bovine aortic endothelial cells (BAECs) following treatment with serum or phorbol ester. Zf9 transcriptionally activates urokinase plasminogen activator (uPA). Recombinant Zf9-GST binds to wild-type but not mutated 'GC-box' motifs within the human uPA promoter (-63 to 32), with greatest affinity to the middle of 3 contiguous GC boxes. Transient transfection of Zf9 drives transactivation of a full-length uPA promoter- and GC box-construct, but not a uPA promoter-construct devoid of GC boxes. Transactivation of uPA by Zf9 is also supported in Drosophila S2 cells. Most importantly, transiently transfected Zf9 up-regulates endogenous uPA messenger RNA and activity in BAECs, resulting in increased bioactive transforming growth factor-beta (TGF-beta) via enhancement of proteolytic activation of the latent molecule. Furthermore, concomitant expression of Zf9 and uPA proteins was observed in arterial endothelial cells after balloon injury in rats, suggesting a potential role of Zf9 in uPA expression not only in vitro but also in vivo. These findings suggest a role of Zf9 in the injury response by enhancing uPA synthesis and subsequent activation of latent TGF-beta. (Blood. 2000;95:1309-1316) PMID- 10666205 TI - Leukocyte-leukocyte interactions mediated by platelet microparticles under flow. AB - Platelet microparticles (PMPs) are released from activated platelets and express functional adhesion receptors, including P-selectin, on their surface. PMP concentrations are elevated in many disorders, and their role in accelerating coagulation has been studied. However, their role in leukocyte aggregation has not been defined. We hypothesized that P-selectin-expressing PMPs bridge leukocytes that express P-selectin glycoprotein ligand-1 (PSGL-1), thereby allowing them to interact under flow conditions. PMPs were isolated from platelet rich plasma or were generated by activating washed platelets with calcium ionophore. PMPs increased transient adhesion of flowing HL-60 cells or neutrophils to HL-60 cells or neutrophils prebound to the surface of a parallel plate flow chamber. Homotypic neutrophil interactions are initiated by the binding of L-selectin to PSGL-1. However, even when L-selectin function was blocked, PMPs allowed flowing neutrophils to aggregate and to interact with PSGL 1-expressing cells prebound to the surface of the flow chamber. The microparticle mediated cell interactions occurred at lower shear stresses than those mediated by L-selectin. PMPs may enhance leukocyte aggregation and leukocyte accumulation on selectin-expressing substrates, especially in diseases where the concentration of the particles is elevated. (Blood. 2000;95:1317-1323) PMID- 10666206 TI - Prevention and treatment of factor VIII inhibitors in murine hemophilia A. AB - Inhibitory antibody formation is a major complication of factor VIII replacement therapy in patients with hemophilia A. To better understand the pathogenesis of this immunologic reaction, we evaluated the role of T-cell costimulatory signals for antifactor VIII antibody formation in a murine model of hemophilia A. Repeated intravenous injections of factor VIII in these factor VIII-deficient mice induced an antifactor VIII inhibitor antibody response. This response was shown to be T-cell dependent by its absence in hemophilic mice also deficient for the T-cell costimulatory ligand B7-2. In separate experiments, injection of murine CTLA4-Ig completely blocked the primary response to factor VIII in hemophilic mice with intact B7 function. This reagent also prevented or diminished further increases in antifactor VIII when given to hemophilic mice with low antifactor VIII antibody titers. These studies suggest that strategies targeting the B7-CD28 pathway are potential therapies to prevent and treat inhibitory antifactor VIII antibodies. Moreover, because the development of antibodies to replaced proteins may limit the success of many human gene therapy approaches, our results may be broadly applicable. (Blood. 2000;95:1324-1329) PMID- 10666207 TI - Inhibition of thrombin generation by the zymogen factor VII: implications for the treatment of hemophilia A by factor VIIa. AB - Factor VII circulates as a single chain inactive zymogen (10 nmol/L) and a trace ( approximately 10-100 pmol/L) circulates as the 2-chain form, factor VIIa. Factor VII and factor VIIa were studied in a coagulation model using plasma concentrations of purified coagulation factors with reactions initiated with relipidated tissue factor (TF). Factor VII (10 nmol/L) extended the lag phase of thrombin generation initiated by 100 pmol/L factor VIIa and low TF. With the coagulation inhibitors TFPI and AT-III present, factor VII both extended the lag phase of the reaction and depressed the rate of thrombin generation. The inhibition of factor Xa generation by factor VII is consistent with its competition with factor VIIa for TF. Thrombin generation with TF concentrations >100 pmol/L was not inhibited by factor VII. At low tissue factor concentrations (<25 pmol/L) thrombin generation becomes sensitive to the absence of factor VIII. In the absence of factor VIII, factor VII significantly inhibits TF-initiated thrombin generation by 100 pmol/L factor VIIa. In this hemophilia A model, approximately 2 nmol/L factor VIIa is needed to overcome the inhibition of physiologic (10 nmol/L) factor VII. At 10 nmol/L, factor VIIa provided a thrombin generation response in the hemophilia model (0% factor VIII, 10 nmol/L factor VII) equivalent to that observed with normal plasma, (100% factor VIII, 10 nmol/L factor VII, 100 pmol/L factor VIIa). These results suggest that the therapeutic efficacy of factor VIIa in the medical treatment of hemophiliacs with inhibitors is, in part, based on overcoming the factor VII inhibitory effect. (Blood. 2000;95:1330-1335) PMID- 10666208 TI - Missense mutations in the human beta fibrinogen gene cause congenital afibrinogenemia by impairing fibrinogen secretion. AB - Congenital afibrinogenemia is a rare autosomal recessive disorder characterized by bleeding that varies from mild to severe and by complete absence or extremely low levels of plasma and platelet fibrinogen. Although several mutations in the fibrinogen genes associated with dysfibrinogenemia and hypofibrinogenemia have been described, the genetic defects of congenital afibrinogenemia are largely unknown, except for a recently reported 11-kb deletion of the fibrinogen Aalpha chain gene. Nevertheless, mutation mechanisms other than the deletion of a fibrinogen gene are likely to exist because patients with afibrinogenemia showing no gross alteration within the fibrinogen cluster have been reported. We tested this hypothesis by studying the affected members of two families, one Italian and one Iranian, who had no evidence of large deletions in the fibrinogen genes. Sequencing of the fibrinogen genes in the 2 probands detected 2 different homozygous missense mutations in exons 7 and 8 of the Bbeta-chain gene, leading to amino acid substitutions Leu353Arg and Gly400Asp, respectively. Transient transfection experiments with plasmids expressing wild-type and mutant fibrinogens demonstrated that the presence of either mutation was sufficient to abolish fibrinogen secretion. These findings demonstrated that missense mutations in the Bbeta fibrinogen gene could cause congenital afibrinogenemia by impairing fibrinogen secretion. (Blood. 2000;95:1336-1341) PMID- 10666209 TI - Stimulation of cytotoxic T cells against idiotype immunoglobulin of malignant lymphoma with protein-pulsed or idiotype-transduced dendritic cells. AB - Because of their hypervariable regions and somatic mutations, the antigen receptor molecules of lymphomas (idiotypes) are tumor-specific antigens and attractive targets for antilymphoma immunotherapy. For the optimal induction of human idiotype-specific cytotoxic T cells (CTL), idiotype was presented to CD8(+) peripheral blood mononuclear cells by monocyte-derived autologous dendritic cells (DC) after the endocytosis of idiotype protein or by idiotype-expressing DC. Recombinant idiotype was obtained as a functionally folded Fab fragment by periplasmic expression in Escherichia coli. Idiotype-expressing DC were generated by transduction with recombinant Semliki forest virus vectors encompassing heavy- or light-chain idiotype genes. Autologous lymphoblastoid cell lines stably transfected with Epstein-Barr virus-based idiotype expression vectors were used as target cells to detect idiotype-specific lysis. CTL stimulated with idiotype loaded DC showed strong specific, CD8-mediated, and major histocompatibility complex (MHC) class I-restricted cytotoxicity against autologous heavy- and light chain idiotype. In contrast, stimulation with idiotype-transduced DC resulted in only moderate natural killer cell activity. These data confirm the existence of idiotype-specific CTL in patients with lymphoma, define a "good manufacturing practice"-compatible protocol for the generation of these cells without the requirement of viable lymphoma cells, and favor the processing of exogenous antigen over DC transduction for the induction of MHC I-restricted CTL against idiotypes with unknown antigenicity. (Blood. 2000;95:1342-1349) PMID- 10666210 TI - Circulating forms of intercellular adhesion molecule (ICAM)-1 in mice lacking membranous ICAM-1. AB - Mice deficient in intercellular adhesion molecule-1 (ICAM-1), lacking membranous ICAM-1, show a normal development but abnormalities of inflammatory and immune functions. Although the membrane-bound form of ICAM-1 is not detectable in the mutant strain, circulating ICAM-1 (cICAM) is present in serum from ICAM-1 deficient mice in similar amounts as in serum from wild-type mice. These findings were confirmed in vitro by flow cytometric analysis of lipopolysaccharide stimulated spleen cells, and cICAM-enzyme-linked immunosorbent assay analysis of supernatants of cultured spleen cells. To analyze for the source of cICAM-1, spleen cell RNA was isolated and ICAM-1 RNA was amplified by reverse transcriptase-polymerase chain reaction using primers binding in the 5' and 3' untranslated regions. Different fragments were cloned and sequenced. In wild-type RNA the common 5 domain form of ICAM-1 was identified. In RNA from ICAM-1 mutant mice only 3 smaller fragments were found. Sequencing these fragments identified 3 alternatively spliced isoforms of ICAM-1, lacking 2 or 3 extracellular domains. However, in all spliced fragments the transmembrane domain was included. Therefore, we postulate that circulating forms of ICAM-1 are generated by proteolytic cleavage of membranous ICAM-1. The data indicate that the expression of membranous ICAM-1 and the appearance of circulating forms in serum are independently regulated mechanisms. (Blood. 2000;95:1350-1355) PMID- 10666211 TI - Thymocyte development past the CD4(+)CD8(+) stage requires an active p38 mitogen activated protein kinase. AB - Activation of the p38 mitogen-activated protein kinase (MAPK) pathway is important for some T-cell functions, but its role in intrathymic development is unclear. To investigate the function of p38 MAPK during the late stages of thymocyte differentiation, pharmacologic and genetic manipulations were used to inhibit p38 MAPK activity in developing thymocytes. Ligation of the T-cell antigen receptor (TCR) on either thymocytes or a thymocyte cell line resulted in p38 MAPK activation. Selective pharmacologic inhibition of p38 MAPK activity with the pyridinyl imidazole drug SB203580 severely impaired the development of mature CD4(+) and CD8(+) single positive (SP) thymocytes from their CD4(+)CD8(+) double positive (DP) precursors in fetal thymic organ culture (FTOC). Further, pharmacologic or genetic suppression of p38 MAPK activity, the latter achieved by overexpressing a catalytically inactive p38 MAPK, resulted in a blockade of the DP-to-SP transition of a thymocyte cell line in a novel in vitro differentiation assay. Taken together, these data constitute the first demonstration that p38 MAPK plays a critical role in the DP-to-SP differentiation of thymocytes during late intrathymic development. (Blood. 2000;95:1356-1361) PMID- 10666212 TI - Migration of activated CD8(+) T lymphocytes to sites of viral infection does not require endothelial selectins. AB - Using mice deficient of E-selectin and E/P-selectin, we have studied the requirement for endothelial selectins in extravasation of leukocytes at sites of viral infection, with major emphasis on the recruitment of virus-specific T(C)1 cells. Lymphocytic choriomeningitis virus (LCMV)-induced meningitis was used as our primary experimental model. Additionally, localized subdermal inflammation and virus clearance in internal organs were analyzed during LCMV infection. The generation of CD8(+) effector T cells in infected mutants was unimpaired. Quantitative and qualitative analysis of the inflammatory exudate cells in intracerebrally infected mice gave identical results in all strains of mice. Expression of endothelial selectin was also found to be redundant regarding the ability of effector cells to eliminate virus in nonlymphoid organs. Concerning LCMV-induced footpad swelling, absent or marginal reduction was found in E/P-sel /- mice, compared with wild-type mice after local challenge with virus or immunodominant viral MHC class I restricted peptide, respectively. Similar results were obtained after adoptive transfer of wild-type effector cells into E/P-sel -/- recipients, whereas footpad swelling was markedly decreased in P sel/ICAM-1 -/- and ICAM-1 -/- recipients. LCMV-induced footpad swelling was completely inhibited in ICAM-deficient mice transfused with donor cell preincubated with soluble VCAM-1-Ig chimeric protein. Taken together, the current findings strongly indicate that the migration of T(C)1 effector cells to sites of viral infection can proceed in the absence of endothelial selectins, whereas ligands of the Ig superfamily are critically involved in this process. (Blood. 2000;95:1362-1369) PMID- 10666213 TI - Both stat5a and stat5b are required for antigen-induced eosinophil and T-cell recruitment into the tissue. AB - Antigen-induced eosinophil recruitment into the airways of sensitized mice is mediated by CD4(+) T cells and their cytokines, especially IL-5. In this study, we found that the antigen-induced airway eosinophilia was diminished in Stat5a deficient (Stat5a(-/-)) mice and Stat5b-deficient (Stat5b(-/-)) mice. We also found that antigen-induced CD4(+) T-cell infiltration and IL-5 production in the airways were diminished in Stat5a(-/- )mice and Stat5b(-/-) mice. Moreover, antigen-induced proliferation of splenocytes was diminished in Stat5a(-/- )mice and Stat5b(-/-) mice, suggesting that the generation of antigen-primed T cells may be compromised in Stat5a(-/-) mice and Stat5b(-/-) mice and this defect may account for the diminished antigen-induced T-cell infiltration into the airways. Interestingly, IL-4 and IL-5 production from anti-CD3-stimulated splenocytes was diminished in Stat5a(-/-) mice and Stat5b(-/-) mice. However, antigen-specific IgE and IgG1 production was diminished in Stat5a(-/-) mice but not in Stat5b(-/-) mice, whereas antigen-specific IgG2a production was increased in Stat5a(-/-) mice, suggesting the enhanced Th1 responses in Stat5a(-/-) mice. Finally, we found that eosinophilopoiesis induced by the administration of recombinant IL-5 was also diminished in Stat5a(-/-) mice and Stat5b(-/-) mice. Together, these results indicate that both Stat5a and Stat5b are essential for induction of antigen-induced eosinophil recruitment into the airways and that the defects in antigen-induced eosinophil recruitment in Stat5a(-/-) mice and Stat5b(-/-) mice result from both impaired IL-5 production in the airways and diminished IL-5 responsiveness of eosinophils. (Blood. 2000;95:1370-1377) PMID- 10666214 TI - Gene expressions of lipopolysaccharide receptors, toll-like receptors 2 and 4, are differently regulated in mouse T lymphocytes. AB - Toll-like receptors (TLRs) are a family of mammalian proteins homologous to Drosophila Toll. Human TLR2 was shown to mediate the responsiveness to lipopolysaccharide (LPS). On the other hand, gene mutations of mouse TLR4 (mTLR4) in LPS-hyporesponsive strains have suggested that mTLR4 is essential for LPS signaling in mice, but the role of mTLR2 has not been explored. This report describes molecular cloning of the mTLR2 cDNA. Overexpression of mTLR2 and mouse CD14 conferred LPS-inducibility of c-Jun N-terminal kinase phosphorylation and nuclear factor-kappaB activation to COS7 cells, suggesting that mTLR2 is a signaling receptor for LPS. Both mTLR2 and mTLR4 genes were expressed in T cells. Treatment with anti-CD3epsilon, PMA plus ionomycin, or interleukin-2 (IL-2)/IL-15 increased mTLR2 but not mTLR4 messenger RNA (mRNA) in some T cell lines. Specific inhibitors of mitogen-activated extracellular signal-regulated kinase and fusion protein 38 (p38) kinase inhibited mTLR2 mRNA up-regulation by PMA plus ionomycin. This suggests that extracellular signal-regulated kinase and p38 kinase pathways were involved. Additionally, LPS treatment of EL-4 cell line decreased IL-4 gene expression. Our results indicate that both mTLR2 and mTLR4 are involved in LPS signaling, but their expressions are regulated differently in T cells, and that LPS may directly affect T-cell functions by binding to TLRs. (Blood. 2000;95:1378 1385) PMID- 10666215 TI - Abundant tax protein expression in CD4+ T cells infected with human T-cell lymphotropic virus type I (HTLV-I) is prevented by cytotoxic T lymphocytes. AB - The role of the cellular immune response in human T-cell leukemia virus type I (HTLV-I) infection is not fully understood. A persistently activated cytotoxic T lymphocyte (CTL) response to HTLV-I is found in the majority of infected individuals. However, it remains unclear whether this CTL response is protective or causes tissue damage. In addition, several observations paradoxically suggest that HTLV-I is transcriptionally silent in most infected cells and, therefore, not detectable by virus-specific CTLs. With the use of a new flow cytometric procedure, we show here that a high proportion of naturally infected CD4+ peripheral blood mononuclear cells (PBMC) (between 10% and 80%) are capable of expressing Tax, the immunodominant target antigen recognized by virus-specific CTLs. Furthermore, we provide direct evidence that autologous CD8+ T cells rapidly kill CD4+ cells naturally infected with HTLV-I and expressing Tax in vitro by a perforin-dependent mechanism. Consistent with these observations, we observed a significant negative correlation between the frequency of Tax(11-19) specific CD8+ T cells and the percentage of CD4+ T cells in peripheral blood of patients infected with HTLV-I. Those results are in accordance with the view that virus-specific CTLs participate in a highly efficient immune surveillance mechanism that persistently destroys Tax-expressing HTLV-I-infected CD4+ T cells in vivo. (Blood. 2000;95:1386-1392) PMID- 10666216 TI - Evidence that immunoglobulin specificities of AIDS-related lymphoma are not directed to HIV-related antigens. AB - Chronic B-cell stimulation may be a predisposing event in the early pathogenesis of the acquired immunodeficiency syndrome (AIDS)-related lymphoma (ARL). ARL derived immunoglobulin (Ig) genes are significantly diversified from germline, suggesting that antigenic stimulation via Ig receptors may occur prior to malignant transformation. We have evaluated 6 ARL-derived antibodies for binding to human immunodeficiency virus (HIV) and cell surface epitopes. Five cases expressed IgM, and 1 case expressed IgG. Expressed V genes were significantly diversified (3%-15%) from known germline V genes. A non-Ig producing mouse myeloma cell line was transfected with expression vectors containing the lymphoma derived V genes. By enzyme-linked immunosorbent assay and Western blot assay, the lymphoma-derived Ig's showed no reactivity against HIV recombinant proteins. Also, no specific HIV reactivity was observed by flow cytometry with lymphoma derived Ig's against the T-cell line infected with T-tropic HIV-1 or peripheral blood mononuclear cells infected with M-tropic HIV strains, indicating lack of binding to native HIV epitopes. However, 2 of the lymphoma-derived Ig's (ARL-7 and ARL-14) bound strongly to non-HIV-infected cells of various tissue origins. Thus, these findings suggest that the transformed B cells of AIDS-associated lymphomas may not arise from the pool of anti-HIV specific B cells but, rather, may develop from B cells responding to other antigens, including self-antigens. (Blood. 2000;95:1393-1399) PMID- 10666217 TI - Mutation analysis of the 5' noncoding regulatory region of the BCL-6 gene in non Hodgkin lymphoma: evidence for recurrent mutations and intraclonal heterogeneity. AB - The BCL-6 proto-oncogene is involved in the genesis of non-Hodgkin lymphoma (NHL). Rearrangements due to chromosomal translocations and somatic mutations of the 5' noncoding regulatory region of the BCL-6 gene are potential mechanisms for altering its expression in NHL. To further elucidate the nature of the somatic mutations in the regulatory region of this gene, we have studied 10 healthy donors and 11 NHL biopsy samples by extensive molecular cloning and sequencing. In addition, we analyzed the BCL-6 genes of tumor and nontumor cells from 2 of the cases. The germ line sequence of this region was defined, which differs in 7 positions from that previously reported. In addition, 1 polymorphic variation at position 397(G or C) was identified. Deletions, insertions, and repeated substitution mutations were detected among the molecular isolates in 8 tumor specimens, with a mutational incidence ranging from 1.3 x 10(-3) to 1.3 x 10( 2)/bp (base pair). A total of 20 distinct substitution mutations, 1 insertion and 3 deletions were observed. One of these deletion mutations and 2 of the substitutions were observed in more than 1 tumor specimen from different individuals. In 3 tumor samples, identical mutations affecting both alleles were observed. These findings suggest the presence of mutational hot spots and hot specific events, a finding supported by our compilation of previously published data. In 6 samples, the nucleotide sequences showed evidence of intraclonal heterogeneity, consistent with a stepwise ongoing mutational process affecting the BCL-6 gene in the tumor cells. These mutations accumulating in the regulatory region of the BCL-6 gene could play a role in lymphoma progression and in the transformation of follicular lymphomas to more aggressive large cell lymphomas. (Blood. 2000;95:1400-1405) PMID- 10666218 TI - HHV-8 is associated with a plasmablastic variant of Castleman disease that is linked to HHV-8-positive plasmablastic lymphoma. AB - Castleman disease (CD) is a lymphoproliferative disorder of unknown etiology that is associated with the development of secondary tumors, including B-cell lymphoma. Human herpesvirus 8 (HHV-8) (Kaposi's sarcoma-associated herpesvirus) sequences have been described in some cases of multicentric Castleman disease (MCD). Using a monoclonal antibody against an HHV-8-latent nuclear antigen, we show that HHV-8 is specifically associated with a variant of MCD in which HHV-8 positive plasmablasts that show lambda light-chain restriction localize in the mantle zone of B-cell follicles and coalesce to form microscopic lymphomas in some cases. Furthermore, we show that the frank plasmablastic lymphoma that develops in patients with this plasmablastic variant of MCD is also positive for HHV-8 and lambda light chain. Plasmablastic lymphoma associated with MCD is a new disease entity associated with HHV-8 infection. (Blood. 2000;95:1406-1412) PMID- 10666219 TI - Analysis of expressed immunoglobulin heavy chain genes in familial B-CLL. AB - In this study, we wished to determine whether familial chronic lymphocytic leukemia of B-cell phenotype (CLL) shares with sporadic B-CLL the same immunoglobulin (Ig) heavy chain variable region (VH) gene usage and occurrence of somatic mutation, to gain insight into the pathogenetic relatedness of these epidemiologically distinct forms of CLL. We therefore analyzed the expressed Ig heavy chain genes in 23 cases (11 families) of familial CLL, and compared these results with data previously reported for sporadic CLL. In addition, we assessed the relationship of the occurrence of somatic mutation to several clinical and phenotypic features. The distribution of V genes among these cases was similar to that observed in sporadic CLL: VH3 > VH1 > VH4. Thirteen of the 23 cases (57%) showed germ line VH gene sequences, whereas somatic mutations were detected in 10 cases (43%). The average mutation frequency of these latter 10 cases of was 6.7% (ranging from 1.7% to 8.8%), and evidence of antigen selection was noted in 6. Intraclonal variation, followed by clonal evolution and the appearance of a second clone over a 20-year period was observed in 1 case, suggesting that mutations can continue to accumulate after neoplastic transformation. The presence of somatic mutations correlated with age at presentation, low white blood cell (WBC) count, and low fluorescence intensity of surface CD5, and the potential significance of these relationships is discussed. Our data indicate that familial and sporadic B-CLL display a similar pattern of immunoglobulin gene usage and frequency of somatic mutation, and are consistent with a common ontogeny and immunogenetic origin for these 2 epidemiologically distinct forms of CLL. (Blood. 2000;95:1413-1419) PMID- 10666220 TI - Thioredoxin prolongs survival of B-type chronic lymphocytic leukemia cells. AB - Thioredoxin (Trx) is a ubiquitous protein disulfide oxidoreductase with antioxidant, cytokine, and chemotactic properties. Previously, we showed that Trx, in synergy with interleukin 1 (IL-1), IL-2, IL-4, tumor necrosis factor alpha (TNF-alpha), and CD40-ligation induced S-phase entry and mitosis in normal B cells and B-type chronic lymphocytic leukemia (B-CLL) cells. The viability of B CLL cells stimulated by these protocols is high, and it has been hypothesized that the overexpression of Bcl-2 found in B-CLL protects the cells from apoptosis in vitro and in vivo. In this study, we have analyzed the response of cells derived from 12 samples of patients with B-CLL to recombinant human Trx in spontaneous apoptosis, with special reference to the Bcl-2 expression. Long-term cultures of B-CLL clones showed significantly higher viability when supplemented with human Trx (P =.031), also exemplified with clones surviving more than 2 months. Short-term cultures of B-CLL cells exposed to 1 microg/mL of Trx for 1, 5, or 12 days maintained expression or delayed down-regulation of Bcl-2 compared with control cultures containing RPMI 1640 medium and 10% fetal calf serum only (P =.032,. 002,.026, respectively). All B-CLL cells expressed constitutive Trx at varying but low levels, in contrast to adult T-cell leukemias, which overexpress Trx, as previously reported. We found that Trx added to B-CLL cells increased in a dose-dependent fashion the release of TNF-alpha, which has been suggested to be an autocrine growth factor for these cells. In conclusion, we have found that human recombinant Trx induced TNF-alpha secretion, maintained Bcl-2, and reduced apoptosis in B-CLL cells. (Blood. 2000;95:1420-1426) PMID- 10666221 TI - Apoptosis of leukemic cells accompanies reduction in intracellular pH after targeted inhibition of the Na(+)/H(+) exchanger. AB - The Na(+)/H(+) exchanger isoform 1 (NHE1) is primarily responsible for the regulation of intracellular pH (pH(i)). It is a ubiquitous, amiloride-sensitive, growth factor-activatable exchanger whose role has been implicated in cell-cycle regulation, apoptosis, and neoplasia. Here we demonstrate that leukemic cell lines and peripheral blood from primary patient leukemic samples exhibit a constitutively and statistically higher pH(i) than normal hematopoietic tissue. We then show that a direct correlation exists between pH(i) and cell-cycle status of normal hematopoietic and leukemic cells. Advantage was taken of this relationship by treating leukemic cells with the Na(+)/H(+) exchanger inhibitor, 5-(N, N-hexamethylene)-amiloride (HMA), which decreases the pH(i) and induces apoptosis. By incubating patient leukemic cells in vitro with pharmacologic doses of HMA for up to 5 hours, we show, using flow cytometry and fluorescent ratio imaging microscopy, that when the pH(i) decreases, apoptosis-measured by annexin V and TUNEL methodologies-rapidly increases so that more than 90% of the leukemic cells are killed. The differential sensitivity exhibited between normal and leukemic cells allows consideration of NHE1 inhibitors as potential antileukemic agents. (Blood. 2000;95:1427-1434) PMID- 10666222 TI - Three differentially expressed survivin cDNA variants encode proteins with distinct antiapoptotic functions. AB - Survivin is a member of the inhibitor of apoptosis protein (IAP) family that is believed to play a role in oncogenesis. To elucidate further its physiologic role(s), we have characterized the murine survivin gene and complementary DNA (cDNA). The structural organization of the survivin gene, located on chromosome 11E2, is similar to that of its human counterpart, both containing 4 exons. Surprisingly, 3 full-length murine survivin cDNA clones were isolated, predicting the existence of 3 distinct survivin proteins. The longest open reading frame, derived from all 4 exons, predicts a 140-amino acid residue protein, survivin(140), similar to human survivin, which contains a single IAP repeat and a COOH-terminal coiled-coil domain that links its function to the cell cycle. A second cDNA, which retains intron 3, predicts the existence of a 121-amino acid protein, survivin(121) that lacks the coiled-coil domain. Removal of exon 2 derived sequences by alternative pre-messenger RNA (mRNA) splicing results in a third 40-amino acid residue protein, survivin(40), lacking the IAP repeat and coiled-coil structure. Predictably, only recombinant survivin(140) and survivin(121) inhibited caspase-3 activity. All 3 mRNA species were variably expressed during development from 7.5 days postcoitum. Of the adult tissues surveyed, thymus and testis accumulated high levels of survivin(140) mRNA, whereas survivin(121)-specific transcripts were detected in all tissues, while those representing survivin(40) were absent. Human counterparts to the 3 survivin mRNA transcripts were identified in a study of human cells and tissues. The presence of distinct isoforms of survivin that are expressed differentially suggests that survivin plays a complex role in regulating apoptosis. (Blood. 2000;95:1435-1442) PMID- 10666223 TI - Hodgkin and reed-sternberg cells represent an expansion of a single clone originating from a germinal center B-cell with functional immunoglobulin gene rearrangements but defective immunoglobulin transcription. AB - Single cell studies aimed at clarifying the nature and clonality of Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin's disease (HD) have so far produced conflicting results. Using an improved single cell procedure, the HRS cells of 25 patients with nodular sclerosing HD lacking B- and T-cell antigens, with and without Epstein-Barr virus infection, were analyzed for the presence of immunoglobulin (Ig) gene rearrangements. One patient with HD developed follicular lymphoma 2 years later. Both lymphomas originated from a common precursor identified as a germinal center B cell. The data show that all but one of the investigated cases harbored rearranged Ig genes, which were clonal in all instances and carried a high load of somatic mutations. The Ig coding capacity was preserved in 18 of the 24 cases (75%) with rearrangements. However, expression of Ig messenger RNA was not detectable in the HRS cells with the exception of Ig kappa light chain expression in some tumor cells of 1 case. The lack of Ig gene transcription in HRS cells was confirmed by analyzing the HD cell lines L428 and KM-H2 in transient transfection experiments. An Ig promoter/enhancer reporter construct showed virtually no activity in these cells compared to 5 control B-cell lines. We conclude that (1) classical HD is a B-cell lymphoma in most instances, (2) HRS cells are clonal without any exception, (3) they are derived from germinal center B-cells that (4) mostly lack crippling mutations but (5) have consistently lost their Ig gene transcription ability, due to functional defects in the Ig gene regulatory elements. (Blood. 2000;95:1443 1450) PMID- 10666224 TI - Hematopoietic-specific expression of MEFV, the gene mutated in familial Mediterranean fever, and subcellular localization of its corresponding protein, pyrin. AB - Familial Mediterranean fever (FMF) is a recessively inherited disorder characterized by recurrent, self-limited attacks of fever and serositis and by infiltration of affected tissues by large numbers of neutrophils. A candidate gene for FMF was identified by positional cloning and named "MEFV." The corresponding protein was named "pyrin." To elucidate the currently unknown function of pyrin, we characterized its tissue distribution, regulation of expression during hematopoietic differentiation, and subcellular localization. Reverse transcription-polymerase chain reaction analysis, followed by hybridization with an internal oligonucleotide, demonstrated expression of MEFV in different populations of peripheral blood cells. Among hematopoietic cell lines, MEFV was almost exclusively expressed in cells of the myeloid lineage. Furthermore, MEFV messenger RNA was strongly expressed within 24 hours of dimethyl sulfoxide-induced granulocytic differentiation of HL-60 cells. Analysis of complementary DNA from human solid tumor-derived cell lines revealed expression of MEFV in several cell lines derived from colon and prostate cancers. Expression of MEFV fused to enhanced green fluorescent protein showed that pyrin localized in distinct patches in the cytoplasm, forming a perinuclear cap. Taken together, MEFV is predominantly expressed in myeloid cells and upregulated during myeloid differentiation, and the corresponding protein, pyrin, is expressed in the cytoplasm. (Blood. 2000;95:1451-1455) PMID- 10666225 TI - Aberrant expression of active leukotriene C(4) synthase in CD16(+) neutrophils from patients with chronic myeloid leukemia. AB - Elevated leukotriene (LT)C(4) synthase activity was observed in peripheral blood granulocyte suspensions from patients with chronic myeloid leukemia (CML). Magnetic cell sorting (MACS) with CD16 monoclonal antibodies (mAbs), which were used to fractionate granulocytes from CML patients and healthy individuals, yielded highly purified suspensions of CD16(+) neutrophils. The purity of these cell fractions was verified by extensive morphologic examination. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses, demonstrating the absence of interleukin-4 messenger RNA (IL-4 mRNA), further confirmed the negligible contamination of eosinophils in these fractions. Notably, purified CML CD16(+) neutrophils from all tested patients transformed exogenous LTA(4) to LTC(4). These cells also produced LTC(4 )after activation with ionophore A23187 or the chemotactic peptide fMet-LeuPhe (N-formylmethionyl-leucyl-phenylalanine). Subcellular fractionation revealed that the enzyme activity was exclusively distributed to the microsomal fraction. Expression of LTC(4) synthase mRNA in CML CD16(+) neutrophils was confirmed by RT-PCR. Furthermore, Western blot analyses consistently demonstrated expression of LTC(4) synthase at the protein level in CML CD16(+) neutrophils, whereas expression of microsomal glutathione S transferase 2 occurred occasionally. Expectedly, LTC(4) synthase activity or expression of the protein could not be demonstrated in CD16(+) neutrophil suspensions from any of the healthy individuals. Instead, these cells, as well as CML CD16(+) neutrophils, transformed LTA(4) to LTB(4). The results indicate that aberrant expression of LTC(4) synthase is a regular feature of morphologically mature CML CD16(+) neutrophils. This abnormality, possibly associated with malignant transformation, can lead to increased LTC(4) synthesis in vivo. Such overproduction may be of pathophysiological relevance because LTC(4 )has been demonstrated to stimulate proliferation of human bone marrow-derived myeloid progenitor cells. (Blood. 2000;95:1456-1464) PMID- 10666226 TI - Extracellular granzyme A, complexed to proteoglycans, is protected against inactivation by protease inhibitors. AB - Granzyme A (GrA) and B (GrB) together with perforin are the main constituents of cytotoxic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. The cytotoxic proteins are released to deliver a lethal hit during contact between the CTL or NK cell and target cell. With the use of an enzyme-linked immunosorbent assay for antigenic levels, we showed in a recent study that plasma of patients with activated CTLs and NK cells contain elevated levels of extracellular GrA. In this study, we determined the form and proteolytic capacity of this extracellular GrA detected in plasma. With the use of various assays, we show that part of the extracellular GrA circulates in the mature conformation and is bound to proteoglycans that protect it against inactivation by protease inhibitors, such as antithrombin III and alpha-2-macroglobulin, whereas another part of GrA circulates as a complex with antithrombin III. Finally, with the use of a novel assay for active GrA, we demonstrate that some plasma samples with high levels of extracellular GrA contain active GrA. These results suggest that various forms of extracellular GrA occur in vivo and that the regulation of GrA activity may be modified by proteoglycans. These data support the notion that granzymes may exert extracellular functions distant from the site of CTL or NK cell interaction with their target cells. (Blood. 2000;95:1465-1472) PMID- 10666227 TI - Tropomyosin isoform 5b is expressed in human erythrocytes: implications of tropomodulin-TM5 or tropomodulin-TM5b complexes in the protofilament and hexagonal organization of membrane skeletons. AB - The human erythrocyte membrane skeleton consists of hexagonal lattices with junctional complexes containing F-actin protofilaments of approximately 33-37 nm in length. We hypothesize that complexes formed by tropomodulin, a globular capping protein at the pointed end of actin filaments, and tropomyosin (TM), a rod-like molecule of approximately 33-35 nm, may contribute to the formation of protofilaments. We have previously cloned the human tropomodulin complementary DNA and identified human TM isoform 5 (hTM5), a product of the gamma-TM gene, as one of the major TM isoforms in erythrocytes. We now identify TM5b, a product of the alpha-TM gene, to be the second major TM isoform. TM5a, the alternatively spliced isoform of the alpha-TM gene, which differs by 1 exon and has a weaker actin-binding affinity, however, is not present. TM4, encoded by the delta-TM gene, is not present either. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hTM5 comigrated with the slower TM major species in erythrocyte membranes, and hTM5b comigrated with the faster TM major species. TM5b, like TM5, binds strongly to tropomodulin, more so than other TM isoforms. The 2 major TM isoforms, therefore, share several common features: They have 248 residues, are approximately 33-35 nm long, and have high affinities toward F-actin and tropomodulin. These common features may be the key to the mechanism by which protofilaments are formed. Tropomodulin-TM5 or tropomodulin-TM5b complexes may stabilize F-actin in segments of approximately 33-37 nm during erythroid terminal differentiation and may, therefore, function as a molecular ruler. TM5 and TM5b further define the hexagonal geometry of the skeletal network and allow actin regulatory functions of TMs to be modulated by tropomodulin. (Blood. 2000;95:1473 1480) PMID- 10666228 TI - Red cell surface changes and erythrophagocytosis in children with severe plasmodium falciparum anemia. AB - Severe anemia is one of the most lethal complications in children infected with Plasmodium falciparum. The pathogenesis of this anemia is not completely understood. Experimental data from malaria-infected humans and animal models suggest that uninfected red cells have a shortened life span. This study looked for changes in the red cell surfaces of children with severe malarial anemia that could explain this accelerated destruction. A prospective case-control study was conducted of children with severe P falciparum anemia (hemoglobin of 5 g/dL or lower) admitted to a large general hospital in western Kenya. Children with severe anemia were compared with children who had symptoms of uncomplicated malaria and with asymptomatic children. Cytofluorometry was used to quantify in vitro erythrophagocytosis and to measure red cell surface immunoglobulin G (IgG) and the complement regulatory proteins CR1, CD55, and CD59. Red cells from patients with severe anemia were more susceptible to phagocytosis and also showed increased surface IgG and deficiencies in CR1 and CD55 compared with controls. Red cell surface CD59 was elevated in cases of severe anemia compared with asymptomatic controls but not as compared with symptomatic controls. The surface of red cells of children with severe P falciparum anemia is modified by the deposition of IgG and alterations in the levels of complement regulatory proteins. These changes could contribute to the accelerated destruction of red cells in these patients by mechanisms such as phagocytosis or complement-mediated lysis. (Blood. 2000;95:1481-1486) PMID- 10666229 TI - Single-tube multiplex PCR-SSCP analysis distinguishes 7 common ABO alleles and readily identifies new alleles. AB - The ABO blood group is clinically the most important blood group system. Elucidation of the molecular basis of the ABO polymorphism allows genotype determination without family studies. Described here is a new method based on the simultaneous amplification by polymerase chain reaction (PCR) of 3 fragments from exon 6, and 5' and 3' ends of exon 7 of the ABO gene, followed by single-strand conformation polymorphism (SSCP) analysis. This multiplex PCR-SSCP protocol allows the well-established base changes at 9 nucleotide positions 261, 297, 467, 526, 646, 657, 681, 1059, and 1096 to be assayed simultaneously so that 7 common alleles (A(1), A(1v), A(2), B, O(1), O(1v), and O(2)) can be distinguished in a single-tube single-lane format. Each allele was characterized by a set of 3 haplotype-specific SSCP patterns. Chinese (n = 125) and white European (n = 98) samples were analyzed, and their genotypes were found consistent with the serologic phenotypes or could be deduced unambiguously. Fifteen samples (2 Chinese and 13 white European) were each found carrying at least 1 rare allele. Most of these alleles were new and some might be generated by intragenic recombination. This technique is the simplest, quickest, and most informative method reported to date and also readily identifies new alleles. (Blood. 2000;95:1487-1492) PMID- 10666230 TI - The IVS4 + 4 A to T mutation of the fanconi anemia gene FANCC is not associated with a severe phenotype in Japanese patients. AB - Fanconi anemia (FA) is an autosomal recessive disease characterized by congenital anomalies, aplastic anemia, and a susceptibility to leukemia. There are at least 8 complementation groups (A through H). Extensive analyses of the FA group C gene FANCC in Western countries revealed that 10% to 15% of FA patients have mutations of this gene. The most common mutation is IVS4 + 4 A to T (IVS4), a splice mutation in intron 4, which has been found only in patients of Ashkenazi Jewish ancestry. When we screened 29 Japanese patients (20 unrelated patients and 4 families) using polymerase chain reaction-single strand conformation polymorphism, we found 8 unrelated patients homozygous for IVS4. This is apparently the first non-Ashkenazi-Jewish population for whom this mutation has been detected. The Ashkenazi Jewish patients homozygous for IVS4 have a severe phenotype, in comparison with other FA patients. Our analyses of Japanese patients indicate no significant difference between IVS4 homozygotes and other patients with regard to severity of a clinical phenotype. Thus, ethnic background may have a significant effect on a clinical phenotype in FA patients carrying the same mutation. (Blood. 2000;95:1493-1498) PMID- 10666231 TI - Glucose-6-phosphate dehydrogenase aveiro: a de novo mutation associated with chronic nonspherocytic hemolytic anemia. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked enzyme abnormality. The clinical phenotype is variable but often predictable from the molecular lesion. Class I variants (the most severe forms of the disease) cluster within exon 10, in a region that, at the protein level, is believed to be involved in dimerization. Here we describe a de novo mutation (C269Y) of a new class I variant (G6PD Aveiro) that maps to exon 8. Mutant and normal alleles were found in both hematopoietic and buccal cells, indicating the presence of mosaicism. The available model of the protein predicts that this lesion lies in proximity to the dimer interface of the molecule. A possible mechanism to explain the severity of the defect is proposed. (Blood. 2000;95:1499-1501) PMID- 10666232 TI - CD20 monoclonal antibody (rituximab) for therapy of Epstein-Barr virus lymphoma after hemopoietic stem-cell transplantation. AB - After bone marrow transplantation (BMT) using T-cell-depleted marrow from an unrelated donor or HLA-mismatched related donor, the risk of developing lymphoproliferative disease associated with the Epstein-Barr virus (EBV) ranges from 1% to 25%. We have shown that administration of donor-derived EBV-specific cytotoxic T lymphocytes (CTL) is effective prophylaxis and treatment for this complication, and we routinely generate CTL for high-risk patients. However, EBV lymphoma can occur in recipients of matched-sibling transplants for whom CTL are unavailable or in patients for whom CTL administration is contraindicated. We report on 3 such patients, who were successfully and safely treated with rituximab, a CD20 monoclonal antibody. The patients remain disease free 7, 8, and 9 months, respectively, after therapy. We conclude that CD20 antibody may be a useful alternative treatment strategy in patients with EBV lymphoma after BMT. (Blood. 2000;95:1502-1505) PMID- 10666233 TI - The exodus subfamily of CC chemokines inhibits the proliferation of chronic myelogenous leukemia progenitors. AB - Chemokines are a family of related proteins that regulate leukocyte infiltration into inflamed tissue and play important roles in disease processes. Among the biologic activities of chemokines is inhibition of proliferation of normal hematopoietic progenitors. However, chemokines that inhibit normal progenitors rarely inhibit proliferation of hematopoietic progenitors from patients with chronic myelogenous leukemia (CML). We and others recently cloned a subfamily of CC chemokines that share similar amino-terminal peptide sequences and a remarkable ability to chemoattract T cells. These chemokines, Exodus-1/LARC/MIP 3alpha, Exodus-2/SLC/6Ckine/TCA4, and Exodus-3/CKbeta11/MIP-3beta, were found to inhibit proliferation of normal human marrow progenitors. The study described here found that these chemokines also inhibited the proliferation of progenitors in every sample of marrow from patients with CML that was tested. This demonstration of consistent inhibition of CML progenitor proliferation makes the 3 Exodus chemokines unique among chemokines. (Blood. 2000;95:1506-1508) PMID- 10666234 TI - Mutations of the E2F4 gene in hematological malignancies having microsatellite instability. AB - Mutations of coding repeats within the E2F4, TGF-betaRII, BAX, IGFIIR, and hMSH3 are critical targets of microsatellite instability (MSI) in many kinds of cancers. We analyzed 9 childhood acute lymphoblastic leukemia (ALL) samples, 5 acute myelocytic leukemia (AML) samples, and 10 adult T-cell leukemia (ATL) samples having MSI to determine whether they had mutations of the E2F4, TGF betaRII, BAX, IGFIIR, and hMSH3 genes. Frameshift mutations were found at trinucleotide repeats within a coding exon of the E2F4 gene in 2 of 10 (20%) ATL samples and 1 of 9 (11%) childhood ALL samples. No mutations were found in the TGF-betaRII, BAX, IGFIIR, and hMSH3 genes. E2F4 is a transcription factor that influences the cell-cycle progression. These results suggest that mutations of the E2F4 gene, presumably caused by an abnormality of one of the DNA repair genes, may play an important role in development of ATL and childhood ALL. (Blood. 2000;95:1509-1510) PMID- 10666235 TI - Active residues and viral substrate cleavage sites of the protease of the birnavirus infectious pancreatic necrosis virus. AB - The polyprotein of infectious pancreatic necrosis virus (IPNV), a birnavirus, is processed by the viral protease VP4 (also named NS) to generate three polypeptides: pVP2, VP4, and VP3. Site-directed mutagenesis at 42 positions of the IPNV VP4 protein was performed to determine the active site and the important residues for the protease activity. Two residues (serine 633 and lysine 674) were critical for cleavage activity at both the pVP2-VP4 and the VP4-VP3 junctions. Wild-type activity at the pVP2-VP4 junction and a partial block (with an alteration of the cleavage specificity) at the VP4-VP3 junction were observed when replacement occurred at histidines 547 and 679. A similar observation was made when aspartic acid 693 was replaced by leucine, but wild-type activity and specificity were found when substituted by glutamine or asparagine. Sequence comparison between IPNV and two birnavirus (infectious bursal disease virus and Drosophila X virus) VP4s revealed that serine 633 and lysine 674 are conserved in these viruses, in contrast to histidines 547 and 679. The importance of serine 633 and lysine 674 is reminiscent of the protease active site of bacterial leader peptidases and their mitochondrial homologs and of the bacterial LexA-like proteases. Self-cleavage sites of IPNV VP4 were determined at the pVP2-VP4 and VP4-VP3 junctions by N-terminal sequencing and mutagenesis. Two alternative cleavage sites were also identified in the carboxyl domain of pVP2 by cumulative mutagenesis. The results suggest that VP4 cleaves the (Ser/Thr)-X-Ala / (Ser/Ala) Gly motif, a target sequence with similarities to bacterial leader peptidases and herpesvirus protease cleavage sites. PMID- 10666236 TI - Interaction of the cauliflower mosaic virus coat protein with the pregenomic RNA leader. AB - Using the yeast three-hybrid system, the interaction of the Cauliflower mosaic virus (CaMV) pregenomic 35S RNA (pgRNA) leader with the viral coat protein, its precursor, and a series of derivatives was studied. The purine-rich domain in the center of the pgRNA leader was found to specifically interact with the coat protein. The zinc finger motif of the coat protein and the preceding basic domain were essential for this interaction. Removal of the N-terminal portion of the basic domain led to loss of specificity but did not affect the strength of the interaction. Mutations of the zinc finger motif abolished not only the interaction with the RNA but also viral infectivity. In the presence of the very acidic C-terminal domain, which is part of the preprotein but is not present in the mature CP, the interaction with the RNA was undetectable. PMID- 10666237 TI - The rice tungro bacilliform virus gene II product interacts with the coat protein domain of the viral gene III polyprotein. AB - Rice tungro bacilliform virus (RTBV) is a plant pararetrovirus whose DNA genome contains four genes encoding three proteins and a large polyprotein. The function of most of the viral proteins is still unknown. To investigate the role of the gene II product (P2), we searched for interactions between this protein and other RTBV proteins. P2 was shown to interact with the coat protein (CP) domain of the viral gene III polyprotein (P3) both in the yeast two-hybrid system and in vitro. Domains involved in the P2-CP association have been identified and mapped on both proteins. To determine the importance of this interaction for viral multiplication, the infectivity of RTBV gene II mutants was investigated by agroinoculation of rice plants. The results showed that virus viability correlates with the ability of P2 to interact with the CP domain of P3. This study suggests that P2 could participate in RTBV capsid assembly. PMID- 10666238 TI - Induction of the cellular E2F-1 promoter by the adenovirus E4-6/7 protein. AB - The adenovirus type 5 (Ad5) E4-6/7 protein interacts directly with different members of the E2F family and mediates the cooperative and stable binding of E2F to a unique pair of binding sites in the Ad5 E2a promoter region. This induction of E2F DNA binding activity strongly correlates with increased E2a transcription when analyzed using virus infection and transient expression assays. Here we show that while different adenovirus isolates express an E4-6/7 protein that is capable of induction of E2F dimerization and stable DNA binding to the Ad5 E2a promoter region, not all of these viruses carry the inverted E2F binding site targets in their E2a promoter regions. The Ad12 and Ad40 E2a promoter regions bind E2F via a single binding site. However, these promoters bind adenovirus induced (dimerized) E2F very weakly. The Ad3 E2a promoter region binds E2F very poorly, even via a single binding site. A possible explanation of these results is that the Ad E4-6/7 protein evolved to induce cellular gene expression. Consistent with this notion, we show that infection with different adenovirus isolates induces the binding of E2F to an inverted configuration of binding sites present in the cellular E2F-1 promoter. Transient expression of the E4-6/7 protein alone in uninfected cells is sufficient to induce transactivation of the E2F-1 promoter linked to chloramphenicol acetyltransferase or green fluorescent protein reporter genes. Further, expression of the E4-6/7 protein in the context of adenovirus infection induces E2F-1 protein accumulation. Thus, the induction of E2F binding to the E2F-1 promoter by the E4-6/7 protein observed in vitro correlates with transactivation of E2F-1 promoter activity in vivo. These results suggest that adenovirus has evolved two distinct mechanisms to induce the expression of the E2F-1 gene. The E1A proteins displace repressors of E2F activity (the Rb family members) and thus relieve E2F-1 promoter repression; the E4-6/7 protein complements this function by stably recruiting active E2F to the E2F-1 promoter to transactivate expression. PMID- 10666239 TI - Bovine herpesvirus 5 glycoprotein E is important for neuroinvasiveness and neurovirulence in the olfactory pathway of the rabbit. AB - Glycoprotein E (gE) is important for full virulence potential of the alphaherpesviruses in both natural and laboratory hosts. The gE sequence of the neurovirulent bovine herpesvirus 5 (BHV-5) was determined and compared with that of the nonneurovirulent BHV-1. Alignment of the predicted amino acid sequences of BHV-1 and BHV-5 gE open reading frames showed that they had 72% identity and 77% similarity. To determine the role of gE in the differential neuropathogenesis of BHV-1 and BHV-5, we have constructed BHV-1 and BHV-5 recombinants: gE-deleted BHV 5 (BHV-5gEDelta), BHV-5 expressing BHV-1 gE (BHV-5gE1), and BHV-1 expressing BHV 5 gE (BHV-1gE5). Neurovirulence properties of these recombinant viruses were analyzed using a rabbit seizure model (S. I. Chowdhury et al., J. Comp. Pathol. 117:295-310, 1997) that distinguished wild-type BHV-1 and -5 based on their differential neuropathogenesis. Intranasal inoculation of BHV-5 gEDelta and BHV 5gE1 produced significantly reduced neurological signs that affected only 10% of the infected rabbits. The recombinant BHV-1gE5 did not invade the central nervous system (CNS). Virus isolation and immunohistochemistry data suggest that these recombinants replicate and spread significantly less efficiently in the brain than BHV-5 gE revertant or wild-type BHV-5, which produced severe neurological signs in 70 to 80% rabbits. Taken together, the results of neurological signs, brain lesions, virus isolation, and immunohistochemistry indicate that BHV-5 gE is important for efficient neural spread and neurovirulence within the CNS and could not be replaced by BHV-1 gE. However, BHV-5 gE is not required for initial viral entry into olfactory pathway. PMID- 10666240 TI - Roscovitine, a specific inhibitor of cellular cyclin-dependent kinases, inhibits herpes simplex virus DNA synthesis in the presence of viral early proteins. AB - We have previously shown that two inhibitors specific for cellular cyclin dependent kinases (cdks), Roscovitine (Rosco) and Olomoucine (Olo), block the replication of herpes simplex virus (HSV). Based on these results, we demonstrated that HSV replication requires cellular cdks that are sensitive to these drugs (L. M. Schang, J. Phillips, and P. A. Schaffer. J. Virol. 72:5626 5637, 1998). We further established that at least two distinct steps in the viral replication cycle require cdks: transcription of immediate-early (IE) genes and transcription of early (E) genes (L. M. Schang, A. Rosenberg, and P. A. Schaffer, J. Virol. 73:2161-2172, 1999). Since Rosco inhibits HSV replication efficiently even when added to infected cells at 6 h postinfection, we postulated that cdks may also be required for viral functions that occur after E gene expression. In the study presented herein, we tested this hypothesis directly by measuring the efficiency of viral replication, viral DNA synthesis, and expression of several viral genes during infections in which Rosco was added after E proteins had already been synthesized. Rosco inhibited HSV replication, and specifically viral DNA synthesis, when the drug was added at the time of release from a 12-h phosphonoacetic acid (PAA)-induced block in viral DNA synthesis. Inhibition of DNA synthesis was not a consequence of inhibition of expression of IE or E genes in that Rosco had no effect on steady-state levels of two E transcripts under the same conditions in which it inhibited viral DNA synthesis. Moreover, viral DNA synthesis was inhibited by Rosco even in the absence of protein synthesis. In a second series of experiments, the replication of four HSV mutants harboring temperature-sensitive mutations in genes essential for viral DNA replication was inhibited when Rosco was added at the time of shift-down from the nonpermissive to the permissive temperature. Viral DNA synthesis was inhibited by Rosco under these conditions, whereas expression of viral E genes was not affected. We conclude that cellular Rosco-sensitive cdks are required for replication of viral DNA in the presence of viral E proteins. This requirement may indicate that HSV DNA synthesis is functionally linked to transcription, which requires cdks, or that both viral transcription and DNA replication, independently, require viral or cellular factors activated by Rosco-sensitive cdks. PMID- 10666241 TI - T-cell receptor-mediated anergy of a human immunodeficiency virus (HIV) gp120 specific CD4(+) cytotoxic T-cell clone, induced by a natural HIV type 1 variant peptide. AB - Human immunodeficiency virus type 1 (HIV-1) infection triggers a cytotoxic T lymphocyte (CTL) response mediated by CD8(+) and perhaps CD4(+) CTLs. The mechanisms by which HIV-1 escapes from this CTL response are only beginning to be understood. However, it is already clear that the extreme genetic variability of the virus is a major contributing factor. Because of the well-known ability of altered peptide ligands (APL) to induce a T-cell receptor (TCR)-mediated anergic state in CD4(+) helper T cells, we investigated the effects of HIV-1 sequence variations on the proliferation and cytotoxic activation of a human CD4(+) CTL clone (Een217) specific for an epitope composed of amino acids 410 to 429 of HIV 1 gp120. We report that a natural variant of this epitope induced a functional anergic state rendering the T cells unable to respond to their antigenic ligand and preventing the proliferation and cytotoxic activation normally induced by the original antigenic peptide. Furthermore, the stimulation of Een217 cells with this APL generated altered TCR-proximal signaling events that have been associated with the induction of T-cell anergy in CD4(+) T cells. Importantly, the APL-induced anergic state of the Een217 T cells could be prevented by the addition of interleukin 2, which restored their ability to respond to their nominal antigen. Our data therefore suggest that HIV-1 variants can induce a state of anergy in HIV-specific CD4(+) CTLs. Such a mechanism may allow a viral variant to not only escape the CTL response but also facilitate the persistence of other viral strains that may otherwise be recognized and eliminated by HIV specific CTLs. PMID- 10666242 TI - Cytoplasm-to-nucleus translocation of a herpesvirus tegument protein during cell division. AB - We have previously shown that the herpes simplex virus tegument protein VP22 localizes predominantly to the cytoplasm of expressing cells. We have also shown that VP22 has the unusual property of intercellular spread, which involves the movement of VP22 from the cytoplasm of these expressing cells into the nuclei of nonexpressing cells. Thus, VP22 can localize in two distinct subcellular patterns. By utilizing time-lapse confocal microscopy of live cells expressing a green fluorescent protein-tagged protein, we now report in detail the intracellular trafficking properties of VP22 in expressing cells, as opposed to the intercellular trafficking of VP22 between expressing and nonexpressing cells. Our results show that during interphase VP22 appears to be targeted exclusively to the cytoplasm of the expressing cell. However, at the early stages of mitosis VP22 translocates from the cytoplasm to the nucleus, where it immediately binds to the condensing cellular chromatin and remains bound there through all stages of mitosis and chromatin decondensation into the G(1) stage of the next cycle. Hence, in VP22-expressing cells the subcellular localization of the protein is regulated by the cell cycle such that initially cytoplasmic protein becomes nuclear during cell division, resulting in a gradual increase over time in the number of nuclear VP22-expressing cells. Importantly, we demonstrate that this process is a feature not only of VP22 expressed in isolation but also of VP22 expressed during virus infection. Thus, VP22 utilizes an unusual pathway for nuclear targeting in cells expressing the protein which differs from the nuclear targeting pathway used during intercellular trafficking. PMID- 10666243 TI - Sensitivity to a nonpeptidic compound (RPR103611) blocking human immunodeficiency virus type 1 Env-mediated fusion depends on sequence and accessibility of the gp41 loop region. AB - The triterpene RPR103611 is an efficient inhibitor of membrane fusion mediated by the envelope proteins (Env, gp120-gp41) of CXCR4-dependent (X4) human immunodeficiency virus type 1 (HIV-1) strains, such as HIV-1(LAI) (LAI). Other X4 strains, such as HIV-1(NDK) (NDK), and CCR5-dependent (R5) HIV-1 strains, such as HIV-1(ADA) (ADA), were totally resistant to RPR103611. Analysis of chimeric LAI NDK Env proteins identified a fragment of the NDK gp41 ectodomain determining drug resistance. A single difference at position 91, leucine in LAI and histidine in NDK, apparently accounted for their sensitivity or resistance to RPR103611. We had previously identified a mutation of isoleucine 84 to serine in a drug escape LAI variant. Both I84 and L91 are located in the "loop region" of gp41 separating the proximal and distal helix domains. Nonpolar residues in this region therefore appear to be important for the antiviral activity of RPR103611 and are possibly part of its target. However, another mechanism had to be envisaged to explain the drug resistance of ADA, since its gp41 loop region was almost identical to that of LAI. Fusion mediated by chimeric Env consisting of LAI gp120 and ADA gp41, or the reciprocal construct, was fully blocked by RPR103611. The gp120-gp41 complex of R5 strains is stable, relative to that of X4 strains, and this stability could play a role in their drug resistance. Indeed, when the postbinding steps of ADA infection were performed under mildly acidic conditions (pH 6.5 or 6.0), a treatment expected to favor dissociation of gp120, we achieved almost complete neutralization by RPR103611. The drug resistance of NDK was partially overcome by preincubating virus with soluble CD4, a gp120 ligand inducing conformational changes in the Env complex. The antiviral efficacy of RPR103611 therefore depends on the sequence of the gp41 loop and the stability of the gp120-gp41 complex, which could limit the accessibility of this target. PMID- 10666244 TI - Biochemical requirements of virus wrapping by the endoplasmic reticulum: involvement of ATP and endoplasmic reticulum calcium store during envelopment of African swine fever virus. AB - Enwrapment by membrane cisternae has emerged recently as a mechanism of envelopment for large enveloped DNA viruses, such as herpesviruses, poxviruses, and African swine fever (ASF) virus. For both ASF virus and the poxviruses, wrapping is a multistage process initiated by the recruitment of capsid proteins onto membrane cisternae of the endoplasmic reticulum (ER) or associated ER-Golgi intermediate membrane compartments. Capsid assembly induces progressive bending of membrane cisternae into the characteristic shape of viral particles, and envelopment provides virions with two membranes in one step. We have used biochemical assays for ASF virus capsid recruitment, assembly, and envelopment to define the cellular processes important for the enwrapment of viruses by membrane cisternae. Capsid assembly on the ER membrane, and envelopment by ER cisternae, were inhibited when cells were depleted of ATP or depleted of calcium by incubation with A23187 and EDTA or the ER calcium ATPase inhibitor, thapsigargin. Electron microscopy analysis showed that cells depleted of calcium were unable to assemble icosahedral particles. Instead, assembly sites contained crescent-shaped and bulbous structures and, in rare cases, empty closed five-sided particles. Interestingly, recruitment of the capsid protein from the cytosol onto the ER membrane did not require ATP or an intact ER calcium store. The results show that following recruitment of the virus capsid protein onto the ER membrane, subsequent stages of capsid assembly and enwrapment are dependent on ATP and are regulated by the calcium gradients present across the ER membrane cisternae. PMID- 10666245 TI - Sint1, a common integration site in SL3-3-induced T-cell lymphomas, harbors a putative proto-oncogene with homology to the septin gene family. AB - The murine retrovirus SL3-3 is a potent inducer of T-cell lymphomas when inoculated into susceptible newborn mice. Previously, DNAs from twenty SL3-3 induced tumors were screened by PCR for provirus integration sites. Two out of 20 tumors demonstrated clonal provirus insertion into a common region. This region has now been isolated and characterized. The region, named SL3-3 integration site 1 (Sint1), maps to the distal end of mouse chromosome 11, corresponding to human chromosome 17q25, and may be identical to a mouse mammary tumor virus integration site in a T-cell lymphoma, Pad3. Two overlapping genomic lambda clones spanning about 35 kb were isolated and used as a starting point for a search for genes in the neighborhood of the virus integration sites. A genomic fragment was used as a hybridization probe to isolate a 3-kb cDNA clone, the expression of which was upregulated in one of two tumors harboring a provirus in Sint1. The cDNA clone is predicted to encode a protein which shows 97.0% identity to a human septin-like protein encoded by a gene which has been found as a fusion partner gene of MLL in an acute myeloid leukemia with a t(11;17)(q23;q25). Together these findings raise the possibility that a proto-oncogene belonging to the septin family, and located about 15 kb upstream of the provirus integration sites, is involved in murine leukemia virus-induced T-cell lymphomagenesis. PMID- 10666247 TI - Characterization and sequencing of prototypic human T-lymphotropic virus type 1 (HTLV-1) from an HTLV-1/2 seroindeterminate patient. AB - Serological screening for human T-lymphotropic virus type 1 (HTLV-1) parallels the standard screening process for human immunodeficiency virus (HIV), in which samples found positive by enzyme-linked immunosorbent assay (ELISA) are confirmed with a modified Western blot procedure. There are a significant number of cases in which HTLV-1/2 ELISA-positive specimens demonstrate an incomplete banding pattern on this Western blot. Individuals providing these atypical antibody responses are categorized as seroindeterminate for HTLV-1/2. Although HTLV-1 genomic sequences are readily detectable in the peripheral blood lymphocytes (PBL) of seropositive individuals, previous studies have repeatedly demonstrated that PBL from the vast majority of HTLV-1/2 seroindeterminate individuals are PCR negative for HTLV-1. As a result, identification of the agent responsible for this indeterminate reactivity has been of interest. We have generated an HTLV-1 positive B-cell line (SI-1 B) from one of these seroindeterminate individuals. Previous screening for HTLV-1 in PBL from this patient had been routinely negative by primary PCR; however, HTLV-1 tax had been periodically detected by nested PCR. DNA sequence data generated with genomic DNA from the SI-1 B cell line and HTLV-1-specific primers demonstrated the presence of a full-length viral genome with >97% homology to the Cosmopolitan form of HTLV-1. A 12-bp deletion was identified in the 3'-gag/5'-prot region, which would predict translation of altered or nonfunctional proteins from these genes. We propose that this HTLV-1/2 seroindeterminate patient is infected with a prototypic form of HTLV-1 at an extremely low viral load and that this finding may explain HTLV-1/2 seroindeterminate reactivity in at least a subset of these individuals. PMID- 10666246 TI - Identification of poly(ADP-ribose) polymerase as a transcriptional coactivator of the human T-cell leukemia virus type 1 Tax protein. AB - Human T-cell leukemia virus type 1 (HTLV-1) encodes a transcriptional activator, Tax, whose activity is believed to contribute significantly to cellular transformation. Tax stimulates transcription from the proviral promoter as well as from promoters for a variety of cellular genes. The mechanism through which Tax communicates to the general transcription factors and RNA polymerase II has not been completely determined. We investigated whether Tax could function directly through the general transcription factors and RNA polymerase II or if other intermediary factors or coactivators were required. Our results show that a system consisting of purified recombinant TFIIA, TFIIB, TFIIE, TFIIF, CREB, and Tax, along with highly purified RNA polymerase II, affinity-purified epitope tagged TFIID, and semipurified TFIIH, supports basal transcription of the HTLV-1 promoter but is not responsive to Tax. Two additional activities were required for Tax to stimulate transcription. We demonstrate that one of these activities is poly(ADP-ribose) polymerase (PARP), a molecule that has been previously identified to be the transcriptional coactivator PC1. PARP functions as a coactivator in our assays at molar concentrations approximately equal to those of the DNA and equal to or less than those of the transcription factors in the assay. We further demonstrate that PARP stimulates Tax-activated transcription in vivo, demonstrating that this biochemical approach has functionally identified a novel target for the retroviral transcriptional activator Tax. PMID- 10666248 TI - Characterization of human CD4(+) T-cell clones recognizing conserved and variable epitopes of the Lassa virus nucleoprotein. AB - T cells must play the major role in controlling acute human Lassa virus infection, because patients recover from acute Lassa fever in the absence of a measurable neutralizing antibody response. T cells alone seem to protect animals from a lethal Lassa virus challenge, because after experimental vaccination no neutralizing antibodies are detectable. In order to study human T-cell reactivity to single Lassa virus proteins, the nucleoprotein (NP) of Lassa virus, strain Josiah, was cloned, expressed in Escherichia coli, and affinity purified. Peripheral blood mononuclear cells (PBMC) obtained from 8 of 13 healthy, Lassa virus antibody-positive individuals living in the Republic of Guinea, western Africa, were found to proliferate in response to the recombinant protein (proliferation index >/=10). PBMC obtained from one individual with a particularly high proliferative response were used to generate 50 NP-specific T cell clones (TCC). For six of these the epitopes were mapped with overlapping synthetic peptides derived from the sequence of the NP. These CD4(+) TCC displayed high specific proliferation and produced mainly gamma interferon upon stimulation with NP. Because variation of up to 15% in the amino acid sequences of the structural proteins of naturally occurring Lassa virus variants has been observed, the reactivity of the TCC with peptides derived from the homologous epitopes of the Nigeria strain of Lassa virus and of the eastern Africa arenavirus Mopeia was tested. With the Nigeria strain of Lassa virus the levels of homology were 100% for two of these epitopes and 85% for three of them, whereas homology with the respective Mopeia epitopes ranged from 92 to 69%. Reactivity of the TCC with peptides derived from the variable epitopes of the Nigeria strain and of Mopeia was reduced or completely abolished. This report shows for the first time that seropositive individuals from areas of endemicity have very strong memory CD4(+) T-cell responses against the NP of Lassa virus, which are partly strain specific and partly cross-reactive with other Lassa virus strains. Our findings may have important implications for the strategy of designing recombinant vaccines against this mainly T-cell-controlled human arenavirus infection. PMID- 10666249 TI - The enhancer I core region contributes to the replication level of hepatitis B virus in vivo and in vitro. AB - Chronic hepatitis B virus (HBV) infection can lead to liver cirrhosis and hepatocellular carcinoma. Long-term interaction of the immune system with the virus results in the selection of escape mutants and viral persistence. In this work we characterize mutations in the enhancer I region isolated prior to liver transplantation from the HBV genomes of 10 patients with chronic HBV infection. The HBV-genomes were sequenced, and the enhancer I region was cloned into luciferase reporter constructs to determine the transcriptional activity. Functional studies were performed by transfecting HBV replication-competent plasmids into hepatoma cells. Analyses of the replication fitness of the mutant strains were conducted by biochemical analysis. In all HBV genomes the enhancer I region was mutated. Most of these mutations resulted in decreased transcriptional activity. The strongest effects were detectable in strains with mutations in the hepatocyte nuclear factor 3 and 4 (HNF3 and HNF4) binding sites of the enhancer I core domain. Replication-competent HBV constructs containing these mutations demonstrated up to 10-fold-reduced levels of virus replication. Before liver transplantation, when the mutant strains were detected in the patients' sera, low HBV DNA levels were found. After transplantation and reinfection with a wild-type virus, the levels of replication were up to 240-fold higher. Our results show that mutations in the enhancer I region of HBV have a major impact on HBV replication. These mutations may also determine the switch from high to low levels of viral replication which is frequently observed during chronic HBV infection. PMID- 10666250 TI - Cellular receptor traffic is essential for productive duck hepatitis B virus infection. AB - We have investigated the mechanism of duck hepatitis B virus (DHBV) entry into susceptible primary duck hepatocytes (PDHs), using mutants of carboxypeptidase D (gp180), a transmembrane protein shown to act as the primary cellular receptor for avian hepatitis B virus uptake. The variant proteins were abundantly produced from recombinant adenoviruses and tested for the potential to functionally outcompete the endogenous wild-type receptor. Overexpression of wild-type gp180 significantly enhanced the efficiency of DHBV infection in PDHs but did not affect ongoing DHBV replication, an observation further supporting gp180 receptor function. A gp180 mutant deficient for endocytosis abolished DHBV infection, indicating endocytosis to be the route of hepadnaviral entry. With further gp180 variants, carrying mutations in the cytoplasmic domain and characterized by an accelerated turnover, the ability of gp180 to function as a DHBV receptor was found to depend on a wild-type-like sorting phenotype which largely avoids transport toward the endolysosomal compartment. Based on these data, we propose a model in which a distinct intracellular DHBV traffic to the endosome, but not beyond, is a prerequisite for completion of viral entry, i.e., for fusion and capsid release. Furthermore, the deletion of the two enzymatically active carboxypeptidase domains of gp180 did not lead to a loss of receptor function. PMID- 10666251 TI - Virus-induced abrogation of transplantation tolerance induced by donor-specific transfusion and anti-CD154 antibody. AB - Treatment with a 2-week course of anti-CD154 antibody and a single transfusion of donor leukocytes (a donor-specific transfusion or DST) permits skin allografts to survive for >100 days in thymectomized mice. As clinical trials of this methodology in humans are contemplated, concern has been expressed that viral infection of graft recipients may disrupt tolerance to the allograft. We report that acute infection with lymphocytic choriomeningitis virus (LCMV) induced allograft rejection in mice treated with DST and anti-CD154 antibody if inoculated shortly after transplantation. Isografts resisted LCMV-induced rejection, and the interferon-inducing agent polyinosinic:polycytidylic acid did not induce allograft rejection, suggesting that the effect of LCMV is not simply a consequence of nonspecific inflammation. Administration of anti-CD8 antibody to engrafted mice delayed LCMV-induced allograft rejection. Pichinde virus also induced acute allograft rejection, but murine cytomegalovirus and vaccinia virus (VV) did not. Injection of LCMV approximately 50 days after tolerance induction and transplantation had minimal effect on subsequent allograft survival. Treatment with DST and anti-CD154 antibody did not interfere with clearance of LCMV, but a normally nonlethal high dose of VV during tolerance induction and transplantation killed graft recipients. We conclude that DST and anti-CD154 antibody induce a tolerant state that can be broken shortly after transplantation by certain viral infections. Clinical application of transplantation tolerance protocols may require patient isolation to facilitate the procedure and to protect recipients. PMID- 10666252 TI - Interactions of viral protein 3CD and poly(rC) binding protein with the 5' untranslated region of the poliovirus genome. AB - The poly(rC) binding protein (PCBP) is a cellular protein required for poliovirus replication. PCBP specifically interacts with two domains of the poliovirus 5' untranslated region (5'UTR), the 5' cloverleaf structure, and the stem-loop IV of the internal ribosome entry site (IRES). Using footprinting analysis and site directed mutagenesis, we have mapped the RNA binding site for this cellular protein within the stem-loop IV domain. A C-rich sequence in a loop at the top of this large domain is required for PCBP binding and is crucial for viral translation. PCBP binds to stem-loop IV RNA with six-times-higher affinity than to the 5' cloverleaf structure. However, the binding of the viral protein 3CD (precursor of the viral protease 3C and the viral polymerase 3D) to the cloverleaf RNA dramatically increases the affinity of PCBP for this RNA element. The viral protein 3CD binds to the cloverleaf RNA but does not interact directly with stem-loop IV nor with other RNA elements of the viral IRES. Our results indicate that the interactions of PCBP with the poliovirus 5'UTR are modulated by the viral protein 3CD. PMID- 10666253 TI - A structured RNA motif is involved in correct placement of the tRNA(3)(Lys) primer onto the human immunodeficiency virus genome. AB - Human immunodeficiency virus type 1 (HIV-1) reverse transcription is primed by the cellular tRNA(3)(Lys) molecule that binds with its 3'-terminal 18 nucleotides to the fully complementary primer-binding site (PBS) on the viral RNA genome. Besides this complementarity, annealing of the primer may be stimulated by additional base-pairing interactions between other parts of the tRNA molecule and viral sequences flanking the PBS. According to the RNA secondary structure model of the HIV-1 leader region, part of the PBS sequence is involved in base pairing to form a small stem-loop structure, termed the U5-PBS hairpin. This hairpin may be involved in the process of reverse transcription. To study the role of the U5 PBS hairpin in the viral replication cycle, we introduced mutations in the U5 region that affect the stability of this structured RNA motif. Stabilization and destabilization of the hairpin significantly inhibited virus replication. Upon prolonged culturing of the virus mutant with the stabilized hairpin, revertant viruses were obtained with additional mutations that restore the thermodynamic stability of the U5-PBS hairpin. The thermodynamic stability of the U5-PBS hairpin apparently has to stay within narrow limits for efficient HIV-1 replication. Transient transfection experiments demonstrated that transcription of the proviral genomes, translation of the viral mRNAs, and assembly of the virions with a normal RNA content is not affected by the mutations within the U5 PBS hairpin. We show that stabilization of the hairpin reduced the amount of tRNA primer that is annealed to the PBS. Destabilization of the hairpin did not affect tRNA annealing, but the viral RNA-tRNA complex was less stable. These results suggest that the U5-PBS hairpin is involved in correct placement of the tRNA primer on the viral genome. The analysis of virus mutants and revertants and the RNA structure probing experiments presented in this study are consistent with the existence of the U5-PBS hairpin as predicted in the RNA secondary structure model. PMID- 10666254 TI - The membrane-proximal stem region of vesicular stomatitis virus G protein confers efficient virus assembly. AB - In this report, we show that the glycoprotein of vesicular stomatitis virus (VSV G) contains within its extracellular membrane-proximal stem (GS) a domain that is required for efficient VSV budding. To determine a minimal sequence in GS that provides for high-level virus assembly, we have generated a series of recombinant DeltaG-VSVs which express chimeric glycoproteins having truncated stem sequences. The recombinant viruses having chimeras with 12 or more membrane-proximal residues of the G stem, and including the G protein transmembrane-cytoplasmic tail domains, produced near-wild-type levels of particles. In contrast, viruses encoding chimeras with shorter or no G-stem sequences produced approximately 10- to 20-fold less. This budding domain when present in chimeric glycoproteins also promoted their incorporation into the VSV envelope. We suggest that the G-stem budding domain promotes virus release by inducing membrane curvature at sites where virus budding occurs or by recruiting condensed nucleocapsids to sites on the plasma membrane which are competent for efficient virus budding. PMID- 10666255 TI - In vitro- and in vivo-generated defective RNAs of satellite panicum mosaic virus define cis-acting RNA elements required for replication and movement. AB - Satellite panicum mosaic virus (SPMV) depends on its helper virus, panicum mosaic virus (PMV), to provide trans-acting proteins for replication and movement. The 824-nucleotide (nt) genome of SPMV possesses an open reading frame encoding a 17.5-kDa capsid protein (CP), which is shown to be dispensable for SPMV replication. To localize cis-acting RNA elements required for replication and movement, a comprehensive set of SPMV cDNA deletion mutants was generated. The results showed that the 263-nt 3' untranslated region (UTR) plus 73 nt upstream of the CP stop codon and the first 16 nt in the 5' UTR are required for SPMV RNA amplification and/or systemic spread. A region from nt 17 to 67 within the 5' UTR may have an accessory role in RNA accumulation, and a fragment bracketing nt 68 to 104 appears to be involved in the systemic movement of SPMV RNA in a host dependent manner. Unexpectedly, defective RNAs (D-RNAs) accumulated de novo in millet plants coinfected with PMV and either of two SPMV mutants: SPMV-91, which is incapable of expressing the 17.5-kDa CP, and SPMV-GUG, which expresses low levels of the 17.5-kDa CP. The D-RNA derived from SPMV-91 was isolated from infected plants and used as a template to generate a cDNA clone. RNA transcripts derived from this 399-nt cDNA replicated and moved in millet plants coinoculated with PMV. The characterization of this D-RNA provided a biological confirmation that the critical RNA domains identified by the reverse genetic strategy are essential for SPMV replication and movement. The results additionally suggest that a potential "trigger" for spontaneous D-RNA accumulation may be associated with the absence or reduced accumulation of the 17.5-kDa SPMV CP. This represents the first report of a D-RNA associated with a satellite virus. PMID- 10666256 TI - Relative sensitivity of hepatitis B virus and other hepatotropic viruses to the antiviral effects of cytokines. AB - We have previously shown that hepatitis B virus (HBV) replication is inhibited noncytopathically in the livers of transgenic mice following injection of HBV specific cytotoxic T lymphocytes (CTLs) or infection with unrelated hepatotropic viruses, including lymphocytic choriomeningitis virus (LCMV) and adenovirus. These effects are mediated by gamma interferon (IFNgamma), tumor necrosis factor alpha (TNFalpha), and IFNalpha/beta. In the present study, we crossed HBV transgenic mice with mice genetically deficient for IFNgamma (IFNgammaKO), the TNFalpha receptor (TNFalphaRKO), or the IFNalpha/beta receptor (IFNalpha/betaRKO) in order to determine the relative contribution of each cytokine to the antiviral effects observed in each of these systems. Interestingly, we showed that HBV replicates in unmanipulated IFNgammaKO and IFNalpha/betaRKO mice at levels higher than those observed in control mice, implying that baseline levels of these cytokines control HBV replication in the absence of inflammation. We also showed that IFNgamma mediates most of the antiviral effect of the CTLs while IFNalpha/beta is primarily responsible for the early inhibitory effect of LCMV and adenovirus on HBV replication. In addition, we showed that the hepatic induction of IFNalpha/beta observed after injection of poly(I. C) is sufficient to inhibit HBV replication and that a similar antiviral effect is achieved by systemic administration of very high doses of IFNalpha. We also compared the relative sensitivity of LCMV and adenovirus to control by IFNgamma, TNFalpha, or IFNalpha/beta in these animals. Importantly, IFNalpha/betaRKO mice, and to a lesser extent IFNgammaKO mice, showed higher hepatic levels of LCMV RNA and adenovirus DNA and RNA than control mice, underscoring the importance of both interferons in controlling these other viral infections as well. PMID- 10666257 TI - Nuclear accumulation of IE62, the varicella-zoster virus (VZV) major transcriptional regulatory protein, is inhibited by phosphorylation mediated by the VZV open reading frame 66 protein kinase. AB - IE62, the major transcriptional activator protein encoded by varicella-zoster virus (VZV), locates to the nucleus when expressed in transfected cells. We show here that cytoplasmic forms of IE62 accumulate in transfected and VZV-infected cells as the result of the protein kinase activity associated with VZV open reading frame 66 (ORF66). Expression of the ORF66 protein kinase but not the VZV ORF47 protein kinase impaired the ability of coexpressed IE62 to transactivate promoter-reporter constructs. IE62 that was coexpressed with the ORF66 protein accumulated predominantly in the cytoplasm, whereas the normal nuclear localization of other proteins was not affected by the ORF66 protein. In cells infected with VZV, IE62 accumulated in the cytoplasm at late times of infection, whereas in cells infected with a VZV recombinant unable to express ORF66 protein (ROka66S), IE62 was completely nuclear. Point mutations introduced into the predicted serine/threonine catalytic domain and ATP binding domain of ORF66 abrogated its ability to influence IE62 nuclear localization, indicating that the protein kinase activity was required. The region of IE62 that was targeted by ORF66 was mapped to amino acids 602 to 733. IE62 peptides containing this region were specifically phosphorylated in cells coexpressing the ORF66 protein kinase and in cells infected with wild-type VZV but were not phosphorylated in cells infected with ROka66S. We conclude that the ORF66 protein kinase phosphorylates IE62 to induce its cytoplasmic accumulation, most likely by inhibiting IE62 nuclear import. PMID- 10666258 TI - The extracellular domain of herpes simplex virus gE is sufficient for accumulation at cell junctions but not for cell-to-cell spread. AB - Herpes simplex virus (HSV) expresses a number of membrane glycoproteins, including gB, gD, and gH/gL, that function in both entry of virus particles and movement of virus from an infected cell to an uninfected cell (cell-to-cell spread). However, a complex of HSV glycoproteins gE and gI (gE/gI) is required for efficient cell-to-cell spread, especially between cells that form extensive cell junctions, yet it is not necessary for entry of extracellular virions. We previously showed that gE/gI has the capacity to localize specifically to cell junctions; the glycoprotein complex was found at lateral surfaces of cells in contact with other cells but not at those lateral surfaces not forming junctions or at apical surfaces. By virtue of these properties, gE/gI is an important molecular handle on the poorly understood process of cell-to-cell spread. Here, we show that the cytoplasmic domain of gE is important for the proper delivery of gE/gI to lateral surfaces of cells. Without this domain, gE/gI is found on the apical surface of epithelial cells, and more uniformly in the cytoplasm, although incorporation into the virion envelope is unaffected. However, even without proper trafficking signals, a substantial fraction of gE/gI retained the capacity to accumulate at cell junctions. Therefore, the extracellular domain of gE can mediate accumulation of gE/gI at cell junctions, if the glycoprotein can be delivered there, probably through interactions with ligands on the opposing cell. The role of phosphorylation of the cytoplasmic domain of gE was also studied. A second mutant HSV type 1 was constructed in which three serine residues that form a casein kinase II phosphorylation site were changed to alanine residues, reducing phosphorylation by 70 to 80%. This mutation did not affect accumulation at cell junctions or cell-to-cell spread. PMID- 10666259 TI - JC virus enters human glial cells by clathrin-dependent receptor-mediated endocytosis. AB - The human polyomavirus JC virus (JCV) is the etiologic agent of a fatal central nervous system (CNS) demyelinating disease known as progressive multifocal leukoencephalopathy (PML). PML occurs predominantly in immunosuppressed patients and has increased dramatically as a result of the AIDS pandemic. The major target cell of JCV infection and lytic replication in the CNS is the oligodendrocyte. The mechanisms by which JCV initiates and establishes infection of these glial cells are not understood. The initial interaction between JCV and glial cells involves virus binding to N-linked glycoproteins containing terminal alpha(2-6) linked sialic acids. The subsequent steps of entry and targeting of the viral genome to the nucleus have not been described. In this report, we compare the kinetics and mechanisms of infectious entry of JCV into human glial cells with that of the related polyomavirus, simian virus 40 (SV40). We demonstrate that JCV, unlike SV40, enters glial cells by receptor-mediated clathrin-dependent endocytosis. PMID- 10666260 TI - Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines. AB - A tetracycline-regulated gene expression system and a panel of novel monoclonal antibodies were used to examine the subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus (HCV) NS3-NS4A complex in inducible cell lines. The NS3 serine protease domain and the full-length NS3 protein expressed in the absence of the NS4A cofactor were diffusely distributed in the cytoplasm and nucleus. Coexpression of NS4A, however, directed NS3 to the endoplasmic reticulum (ER) or an ER-like modified compartment, as demonstrated by colocalization with 3,3'-dihexyloxacarbocyanine iodide, protein disulfide isomerase, and calnexin, as well as subcellular fractionation analyses. In addition, coexpression with NS4A dramatically increased the intracellular stability of NS3 (mean protein half-life of 26 versus 3 h) and allowed for NS4A dependent trans-cleavage at the NS4B-NS5A junction. Deletion analyses revealed that the hydrophobic amino-terminal domain of NS4A was required for ER targeting of NS3. These results demonstrate the importance of studying HCV proteins in their biological context and define a well-characterized cell culture system for further analyses of the NS3-NS4A complex and the evaluation of novel antiviral strategies against hepatitis C. PMID- 10666261 TI - Host sequences flanking the human T-cell leukemia virus type 1 provirus in vivo. AB - Human pathogenic retroviruses do not have common loci of integration. However, many factors, such as chromatin structure, transcriptional activity, DNA-protein interaction, CpG methylation, and nucleotide composition of the target sequence, may influence integration site selection. These features have been investigated by in vitro integration reactions or by infection of cell lines with recombinant retroviruses. Less is known about target choice for integration in vivo. The present study was conducted in order to assess the characteristics of cellular sequences targeted for human T-cell leukemia virus type 1 (HTLV-1) integration in vivo. Sequencing integration sites from >/=200 proviruses (19 kb of sequence) isolated from 29 infected individuals revealed that HTLV-1 integration is not random at the level of the nucleotide sequence. The virus was found to integrate in A/T-rich regions with a weak consensus sequence at positions within and without of the hexameric repeat generated during integration. These features were not associated with a preference for integration near active regions or repeat elements of the host chromosomes. Most or all of the regions of the genome appear to be accessible to HTLV-1 integration. As with integration in vitro, integration specificity in vivo seems to be determined by local features rather than by the accessibility of specific regions. PMID- 10666262 TI - Most retroviral recombinations occur during minus-strand DNA synthesis. AB - Retroviral RNA molecules are plus, or sense in polarity, equivalent to mRNA. During reverse transcription, the first strand of the DNA molecule synthesized is minus-strand DNA. After the minus strand is polymerized, the plus-strand DNA is synthesized using the minus-strand DNA as the template. In this study, a helper cell line that contains two proviruses with two different mutated gfp genes was constructed. Recombination between the two frameshift mutant genes resulted in a functional gfp. If recombination occurs during minus-strand DNA synthesis, the plus-strand DNA will also contain the functional sequence. After the cell divides, all of its offspring will be green. However, if recombination occurs during plus-strand DNA synthesis, then only the plus-strand DNA will contain the wild-type gfp sequence and the minus-strand DNA will still carry the frameshift mutation. The double-stranded DNA containing this mismatch was subsequently integrated into the host chromosomal DNA of D17 cells, which were unable to repair the majority of mismatches within the retroviral double-strand DNA. After the cell divided, one daughter cell contained the wild-type gfp sequence and the other daughter cell contained the frameshift mutation in the gfp sequence. Under fluorescence microscopy, half the cells in the offspring were green and the other half of the cells were colorless or clear. Thus, we demonstrated that more than 98%, if not all, retroviral recombinations occurred during minus-strand DNA synthesis. PMID- 10666263 TI - Rotavirus infection induces an increase in intracellular calcium concentration in human intestinal epithelial cells: role in microvillar actin alteration. AB - Rotaviruses, which infect mature enterocytes of the small intestine, are recognized as the most important cause of viral gastroenteritis in young children. We have previously reported that rotavirus infection induces microvillar F-actin disassembly in human intestinal epithelial Caco-2 cells (N. Jourdan, J. P. Brunet, C. Sapin, A. Blais, J. Cotte-Laffitte, F. Forestier, A. M. Quero, G. Trugnan, and A. L. Servin, J. Virol. 72:7228-7236, 1998). In this study, to determine the mechanism responsible for rotavirus-induced F-actin alteration, we investigated the effect of infection on intracellular calcium concentration ([Ca(2+)](i)) in Caco-2 cells, since Ca(2+) is known to be a determinant factor for actin cytoskeleton regulation. As measured by quin2 fluorescence, viral replication induced a progressive increase in [Ca(2+)](i) from 7 h postinfection, which was shown to be necessary and sufficient for microvillar F-actin disassembly. During the first hours of infection, the increase in [Ca(2+)](i) was related only to an increase in Ca(2+) permeability of plasmalemma. At a late stage of infection, [Ca(2+)](i) elevation was due to both extracellular Ca(2+) influx and Ca(2+) release from the intracellular organelles, mainly the endoplasmic reticulum (ER). We noted that at this time the [Ca(2+)](i) increase was partially related to a phospholipase C (PLC)-dependent mechanism, which probably explains the Ca(2+) release from the ER. We also demonstrated for the first time that viral proteins or peptides, released into culture supernatants of rotavirus-infected Caco-2 cells, induced a transient increase in [Ca(2+)](i) of uninfected Caco-2 cells, by a PLC-dependent efflux of Ca(2+) from the ER and by extracellular Ca(2+) influx. These supernatants induced a Ca(2+) dependent microvillar F-actin alteration in uninfected Caco-2 cells, thus participating in rotavirus pathogenesis. PMID- 10666264 TI - Characterization of the coronavirus mouse hepatitis virus strain A59 small membrane protein E. AB - The small envelope (E) protein has recently been shown to play an essential role in the assembly of coronaviruses. Expression studies revealed that for formation of the viral envelope, actually only the E protein and the membrane (M) protein are required. Since little is known about this generally low-abundance virion component, we have characterized the E protein of mouse hepatitis virus strain A59 (MHV-A59), an 83-residue polypeptide. Using an antiserum to the hydrophilic carboxy terminus of this otherwise hydrophobic protein, we found that the E protein was synthesized in infected cells with similar kinetics as the other viral structural proteins. The protein appeared to be quite stable both during infection and when expressed individually using a vaccinia virus expression system. Consistent with the lack of a predicted cleavage site, the protein was found to become integrated in membranes without involvement of a cleaved signal peptide, nor were any other modifications of the polypeptide observed. Immunofluorescence analysis of cells expressing the E protein demonstrated that the hydrophilic tail is exposed on the cytoplasmic side. Accordingly, this domain of the protein could not be detected on the outside of virions but appeared to be inside, where it was protected from proteolytic degradation. The results lead to a topological model in which the polypeptide is buried within the membrane, spanning the lipid bilayer once, possibly twice, and exposing only its carboxy terminal domain. Finally, electron microscopic studies demonstrated that expression of the E protein in cells induced the formation of characteristic membrane structures also observed in MHV-A59-infected cells, apparently consisting of masses of tubular, smooth, convoluted membranes. As judged by their colabeling with antibodies to E and to Rab-1, a marker for the intermediate compartment and endoplasmic reticulum, the E protein accumulates in and induces curvature into these pre-Golgi membranes where coronaviruses have been shown earlier to assemble by budding. PMID- 10666265 TI - Production of recombinant snakehead rhabdovirus: the NV protein is not required for viral replication. AB - Snakehead rhabdovirus (SHRV) affects warm water fish in Southeast Asia and belongs to the genus Novirhabdovirus by virtue of its nonvirion gene (NV). Because SHRV grows best at temperatures between 28 and 31 degrees C, we were able to use the T7 expression system to produce viable recombinant SHRV from a cloned cDNA copy of the viral genome. Expression of a positive-strand RNA copy of the 11, 550-nucleotide SHRV genome along with the viral nucleocapsid (N), phosphoprotein (P), and polymerase (L) proteins resulted in the generation of infectious SHRV in cells preinfected with a vaccinia virus vector for T7 polymerase expression. Recombinant virus production was verified by detection of a unique restriction site engineered into the SHRV genome between the NV and L genes. Since we were now able to begin examining the function of the NV gene, we constructed a recombinant virus containing a nonsense mutation located 22 codons into the coding sequence of the NV protein. The NV knockout virus was produced at a concentration as high as that of wild-type virus in cultured fish cells, and the resulting virions appeared to be identical to the wild-type virions in electron micrographs. These initial studies suggest that NV has no critical function in SHRV replication in cultured fish cells. PMID- 10666266 TI - Human T-cell leukemia virus type 1 Tax shuttles between functionally discrete subcellular targets. AB - Human T-cell leukemia virus type 1 (HTLV-1) Tax is a nuclear protein with striking pleiotropic functionality. We recently demonstrated that Tax localizes to a multicomponent nuclear speckled structure (Tax speckled structure [TSS]). Here, we examine these structures further and identify a partial overlap of TSS with transcription hot spots. We used a strategy of directed expression via fusion proteins to determine if these transcription sites are the subtargets within TSS required for Tax function. When fused to human immunodeficiency virus type 1 (HIV-1) Tat, the resulting Tat-Tax fusion protein displayed neither a Tat like nor a Tax-like pattern but rather was targeted specifically to the transcription subsites. The Tat-Tax fusion was able to activate both the HIV-1 long terminal repeat (LTR) and the HTVL-1 LTR at the same level as the individual component; thus, targeting proteins to transcription hot spots was compatible with both Tax and Tat transcription function. In contrast, the fusion with HIV-1 Rev, Rev-Tax, resulted in a pattern of expression that was largely Rev-like (nucleolar and cytoplasmic). The reduced localization of Rev-Tax to transcription sites was reflected in a 10-fold drop in activation of the HTLV-1 LTR. However, there was no loss in the ability of Tax to activate via NF-kappaB. Thus, NF kappaB-dependent Tax function does not require targeting of Tax to these transcription sites and suggests that activation via NF-kappaB is a cytoplasmic function. Selective mutation of the nuclear localization signal site in the Rev portion resulted in retargeting of Rev-Tax to TSS and subsequent restoration of transcription function, demonstrating that inappropriate localization preceded loss of function. Mutation of the nuclear export signal site in the Rev portion had no effect on transcription, although the relative amount of Rev-Tax in the cytoplasm was reduced. Finally, in explaining how Tax can occupy multiple subcellular sites, we show that Tax shuttles from the nucleus to the cytoplasm in a heterokaryon fusion assay. Thus, pleiotropic functionality by Tax is regulatable via shuttling between discrete subcellular compartments. PMID- 10666267 TI - Split-intron retroviral vectors: enhanced expression with improved safety. AB - The inclusion of retrovirus-derived introns within retrovirus-based expression vectors leads to a fraction of the resulting transcripts being spliced. Such splicing has been shown to markedly improve expression (W. J. Krall et al., Gene Ther. 3:37-48, 1996). One way to improve upon this still further might involve the use of more efficient introns instead of those from the provirus. Currently, however, incorporation of such introns remains self-defeating since they are removed in the nucleus of the producer cell. In the past, elaborate ways to overcome this problem have included the use of alphaviruses to make the vector transcripts within the cytoplasm, thus avoiding the nuclear splicing machinery during vector production (K. J. Li and H. Garoff, Proc. Natl. Acad. Sci. USA 95:3650-3654, 1998). We now present a novel design for the inclusion of introns within a retroviral vector. In essence, this is achieved by exploiting the retroviral replication process to copy not only the U3 promoter but also a synthetic splice donor to the 5'-long-terminal-repeat position during reverse transcription. Once copied, synthesized transcripts then contain a splice donor at their 5' end capable of interacting with a consensus splice acceptor engineered downstream of the packaging signal. Upon transduction, we demonstrate these vectors to produce enhanced expression from near fully spliced (and thus packaging signal minus) transcripts. The unique design of these high titer and high-expression retroviral vectors may be of use in a number of gene therapy applications. PMID- 10666268 TI - A chimeric protein containing the N terminus of the adeno-associated virus Rep protein recognizes its target site in an in vivo assay. AB - The Rep78 and Rep68 proteins of adeno-associated virus (AAV) type 2 are involved in DNA replication, regulation of gene expression, and targeting site-specific integration. They bind to a specific Rep recognition sequence (RRS) found in both the viral inverted terminal repeats and the AAVS1 integration locus on human chromosome 19. Previous in vitro studies implied that an N-terminal segment of Rep is involved in DNA recognition, while additional domains might stabilize binding and mediate multimerization. In order to define the minimal requirements for Rep to recognize its target site in the human genome, we developed one-hybrid assays in which DNA-protein interactions are detected in vivo. Chimeric proteins consisting of the N terminus of Rep fused to different oligomerization motifs and a transcriptional activation domain were analyzed for oligomerization, DNA binding, and activation of reporter gene expression. Expression of reporter genes was driven from RRS motifs cloned upstream of minimal promoters and examined in mammalian cells from transfected plasmids and in Saccharomyces cerevisiae from a reporter cassette integrated into the yeast genome. Our results show for the first time that chimeric proteins containing the amino-terminal 244 residues of Rep are able to target the RRS in vitro and in vivo when incorporated into artificial multimers. These studies suggest that chimeric proteins may be used to harness the unique targeting feature of AAV for gene therapy applications. PMID- 10666269 TI - A stable HeLa cell line that inducibly expresses poliovirus 2A(pro): effects on cellular and viral gene expression. AB - A HeLa cell clone (2A7d) that inducibly expresses the gene for poliovirus protease 2A (2A(pro)) under the control of tetracycline has been obtained. Synthesis of 2A(pro) induces severe morphological changes in 2A7d cells. One day after tetracycline removal, cells round up and a few hours later die. Poliovirus 2A(pro) cleaves both forms of initiation factor eIF4G, causing extensive inhibition of capped-mRNA translation a few hours after protease induction. Methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone, a selective inhibitor of 2A(pro), prevents both eIF4G cleavage and inhibition of translation but not cellular death. Expression of 2A(pro) still allows both the replication of poliovirus and the translation of mRNAs containing a picornavirus leader sequence, while vaccinia virus replication is drastically inhibited. Translation of transfected capped mRNA is blocked in 2A7d-On cells, while luciferase synthesis from a mRNA bearing a picornavirus internal ribosome entry site (IRES) sequence is enhanced by the presence of 2A(pro). Moreover, synthesis of 2A(pro) in 2A7d cells complements the translational defect of a poliovirus 2A(pro) defective variant. These results show that poliovirus 2A(pro) expression mimics some phenotypical characteristics of poliovirus-infected cells, such as cell rounding, inhibition of protein synthesis and enhancement of IRES-driven translation. This cell line constitutes a useful tool to further analyze 2A(pro) functions, to complement poliovirus 2A(pro) mutants, and to test antiviral compounds. PMID- 10666270 TI - Clustered charge-to-alanine mutagenesis of the vaccinia virus H5 gene: isolation of a dominant, temperature-sensitive mutant with a profound defect in morphogenesis. AB - The vaccinia virus H5 gene encodes a 22.3-kDa phosphoprotein that is expressed during both the early and late phases of viral gene expression. It is a major component of virosomes and has been implicated in viral transcription and, as a substrate of the B1 kinase, may participate in genome replication. To enable a genetic analysis of the role of H5 during the viral life cycle, we used clustered charge-to-alanine mutagenesis in an attempt to create a temperature-sensitive (ts) virus with a lesion in the H5 gene. Five mutant viruses were isolated, with one of them, tsH5-4, having a strong ts phenotype as assayed by plaque formation and measurements of 24-h viral yield. Surprisingly, no defects in genome replication or viral gene expression were detected at the nonpermissive temperature. By electron microscopy, we observed a profound defect in the early stages of virion morphogenesis, with arrest occurring prior to the formation of crescent membranes or immature particles. Nonfunctional, "curdled" virosomes were detected in tsH5-4 infections at the nonpermissive temperature. These structures appeared to revert to functional virosomes after a temperature shift to permissive conditions. We suggest an essential role for H5 in normal virosome formation and the initiation of virion morphogenesis. By constructing recombinant genomes containing two H5 alleles, wild type and H5-4, we determined that H5-4 exerted a dominant phenotype. tsH5-4 is the first example of a dominant ts mutant isolated and characterized in vaccinia virus. PMID- 10666271 TI - The decreased replicative capacity of simian immunodeficiency virus SIVmac239Delta(nef) is manifest in cultures of immature dendritic cellsand T cells. AB - Transmission of simian immunodeficiency virus SIVmac239Delta(nef) (Delta(nef)) to macaques results in attenuated replication of the virus in most animals and ultimately induces protection against challenge with some pathogenic, wild-type SIV strains. It has been difficult, however, to identify a culture system in which the replication of Delta(nef) is severely reduced relative to that of the wild type. We have utilized a primary culture system consisting of blood-derived dendritic cells (DCs) and autologous T cells. When the DCs were fully differentiated or mature, the DC-CD4(+) T-cell mixtures supported replication of both the parental SIV strain, 239 (the wild type), and its mutant with nef deleted (Delta(nef)), irrespective of virus dose and the cell type introducing the virus to the coculture. In contrast, when immature DCs were exposed to Delta(nef) and cocultured with T cells, virus replication was significantly lower than that of the wild type. Activation of the cultures with a superantigen allowed both Delta(nef) and the wild type to replicate comparably in immature DC T-cell cultures. Immature DCs, which, it has been hypothesized, capture and transmit SIV in vivo, are deficient in supporting replication of Delta(nef) in vitro and may contribute to the reduced pathogenicity of Delta(nef) in vivo. PMID- 10666272 TI - Herpes simplex virus type 1-specific cytotoxic T-lymphocyte arming occurs within lymph nodes draining the site of cutaneous infection. AB - Various studies have shown that major histocompatibility complex class I restricted cytotoxic T lymphocytes (CTL) can be isolated from lymph nodes draining sites of cutaneous infection with herpes simplex virus type 1 (HSV-1). Invariably, detection of this cytolytic activity appeared to require some level of in vitro culture of the isolated lymph node cells, usually for 3 days, in the absence of exogenous viral antigen. This in vitro "resting" period was thought to represent the phase during which committed CD8(+) T cells become "armed" killers after leaving the lymph nodes and prior to their entry into infected tissue as effector CTL. In this study we reexamined the issue of CTL appearance in the HSV 1 immune response and found that cytolytic activity can be isolated directly from draining lymph nodes, although at levels considerably below those found after in vitro culture. By using T-cell receptor elements that represent effective markers for class I-restricted T cells specific for an immunodominant glycoprotein B (gB) determinant from HSV-1, we show that the increase in cytotoxicity apparent after in vitro culture closely mirrors the expansion of gB-specific CTL during the same period. Taken together, our results suggest that HSV-1-specific CTL priming does not appear to require any level of cytolytic machinery arming outside the lymph node compartment despite the absence of any detectable infection within that site. PMID- 10666273 TI - Humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle. AB - Adeno-associated virus (AAV) is being developed as a vector capable of conferring long-term gene expression, which is useful in the treatment of chronic diseases. In most therapeutic applications, it is necessary to readminister the vector. This study characterizes the humoral immune response to AAV capsid proteins following intramuscular injection and its impact on vector readministration. Studies of mice and rhesus monkeys demonstrated the formation of neutralizing antibodies to AAV capsid proteins that persisted for over 1 year and then diminished, but this did not prevent the efficacy of vector readministration. More-detailed studies strongly suggested that the B-cell response was T cell dependent. This was further evaluated with a blocking antibody to human CD4, primatized for clinical trials, in a biologically compatible mouse in which the endogenous murine CD4 gene was functionally replaced with the human counterpart. Transient pharmacologic inhibition of CD4 T cells with CD4 antibody prevented an antivector response long after the effects of the CD4 antibody diminished; readministration of vector without diminution of gene expression was possible. Our studies suggest that truly durable transgene expression (i.e., prolonged genetic engraftment together with vector readministration) is possible with AAV in skeletal muscle, although it will be necessary to transiently inhibit CD4 T cell function to avoid the activation of memory B cells. PMID- 10666274 TI - Role of maternal antibody in natural infection of Peromyscus maniculatus with Sin Nombre virus. AB - Data from naturally infected deer mice (Peromyscus maniculatus) were used to investigate vertical transmission of Sin Nombre virus (SNV) and SNV-specific antibody. The antibody prevalence in juvenile mice (14 g or less) was inversely proportional to the mass of the animal, with juvenile deer mice weighing less than 11 g most likely to be antibody positive (26.9%) and juvenile mice weighing between 13 and 14 g least likely to be antibody positive (12.9%). Although a significant sex bias in seropositivity was detected in adult deer mice, no significant sex bias in seropositivity was detected in juvenile animals. Ten juvenile deer mice were identified that had initially tested positive for SNV specific immunoglobulin G (IgG) by enzyme-linked immunosorbent assay (ELISA) but had subsequently tested negative when recaptured as adults. SNV RNA was detected by reverse transcriptase PCR (RT-PCR) in the blood of ELISA-positive adult deer mice but not in the blood of ELISA-positive juveniles. One of the juvenile mice initially tested negative for SNV RNA but later tested positive when recaptured as an ELISA-positive adult. The RT-PCR results for that individual correlated with the disappearance and then reappearance of SNV-specific IgG, indicating that the presence of SNV RNA at later time points was due to infection with SNV via horizontal transmission. SNV-specific antibody present in both ELISA-positive juvenile and adult mice was capable of neutralizing SNV. Additionally, our data indicate that SNV is not transmitted vertically. PMID- 10666275 TI - The in vitro-synthesized RNA from a cDNA clone of hepatitis E virus is infectious. AB - Hepatitis E virus (HEV) is an important etiological agent of epidemic and sporadic hepatitis, which is endemic to the Indian subcontinent and prevalent in most of the developing parts of the world. The infection is often associated with acute liver failure and high mortality, particularly in pregnant women. In order to develop methods of intervention, it is essential to understand the biology of the virus. This is particularly important as no reliable in vitro culture system is available. We have constructed a cDNA clone encompassing the complete HEV genome from independently characterized subgenomic fragments of an Indian epidemic isolate. Transfection studies were carried out with HepG2 cells using in vitro-transcribed RNA from this full-length HEV cDNA clone. The presence of negative-sense RNA, indicative of viral replication, was demonstrated in the transfected cells by strand-specific reverse transcription-PCR and slot blot hybridization. The viral proteins pORF2 and pORF3 and processed components of the pORF1 polyprotein (putative methyltransferase, helicase, and RNA-dependent RNA polymerase) were identified in the transfected cells by metabolic pulse-labeling with [(35)S]methionine-cysteine, followed by immunoprecipitation with respective antibodies. The expression of viral proteins in the transfected cells was also demonstrated by immunofluorescence microscopy. Viral replication was detected in the transfected cells up to 33 days posttransfection (six passages). The culture supernatant from the transfected cells was able to produce HEV infection in a rhesus monkey (Macaca mulatta) following intravenous injection, indicating the generation of viable HEV particles following transfection of cells with in vitro synthesized genomic RNA. This transient cell culture model using in vitro transcribed RNA should facilitate our understanding of HEV biology. PMID- 10666276 TI - Disulfide bonds and membrane topology of the vaccinia virus A17L envelope protein. AB - The envelope protein encoded by the vaccinia virus A17L open reading frame is essential for virion assembly. Our mutagenesis studies indicated that cysteines 101 and 121 form an intramolecular disulfide bond and that cysteine 178 forms an intermolecular disulfide linking two A17L molecules. This arrangement of disulfide bonds has important implications for the topology of the A17L protein and supports a two-transmembrane model in which cysteines 101 and 121 are intraluminal and cysteine 178 is cytoplasmic. The structure of the A17L protein, however, was not dependent on these disulfide bonds, as a recombinant vaccinia virus with all three cysteine codons mutated to serines retained infectivity. PMID- 10666277 TI - Elevated serum transforming growth factor beta1 levels in Epstein-Barr virus associated diseases and their correlation with virus-specific immunoglobulin A (IgA) and IgM. AB - Transforming growth factor beta (TGF-beta) is an immunosuppressive cytokine which can induce immunoglobulin A (IgA) switch and Epstein-Barr virus (EBV) replication in latently infected cells. Here we report elevated serum levels of TGF-beta in various EBV-associated diseases correlating positively with EBV-specific IgA titers and negatively with IgM titers, suggesting a role for this cytokine in the pathogenesis of these diseases. PMID- 10666278 TI - Contributions of Fas-Fas ligand interactions to the pathogenesis of mouse hepatitis virus in the central nervous system. AB - The pathogenesis of the neurotropic strain of mouse hepatitis virus in Fas deficient mice suggested that Fas-mediated cytotoxicity may be required during viral clearance after the loss of perforin-mediated cytotoxicity. The absence of both Fas- and perforin-mediated cytolysis resulted in an uncontrolled infection, suggesting a redundancy of cytolytic pathways to control virus replication. PMID- 10666279 TI - Epstein-Barr virus entry utilizing HLA-DP or HLA-DQ as a coreceptor. AB - Epstein-Barr virus (EBV) glycoprotein gp350/gp220 association with cellular CD21 facilitates virion attachment to B lymphocytes. Membrane fusion requires the additional interaction between virion gp42 and cellular HLA-DR. This binding is thought to catalyze membrane fusion through a further association with the gp85 gp25 (gH-gL) complex. Cell lines expressing CD21 but lacking expression of HLA class II molecules are resistant to infection by a recombinant EBV expressing enhanced green fluorescent protein. Surface expression of HLA-DR, HLA-DP, or HLA DQ confers susceptibility to EBV infection on resistant cells that express CD21. Therefore, HLA-DP or HLA-DQ can substitute for HLA-DR and serve as a coreceptor in EBV entry. PMID- 10666280 TI - Hepadnavirus envelope topology: insertion of a loop region in the membrane and role of S in L protein translocation. AB - A unique feature of the large hepadnavirus envelope protein (L) is its mixed transmembrane topology, resulting from partial posttranslational translocation of the pre-S domain. Using protease protection analysis, we demonstrate for duck hepatitis B virus an essential role for the small envelope protein (S) in this process, providing the first experimental evidence for an S translocation channel. Further analysis revealed that the presumed cytoplasmic loop between TM1 and TM2 in the C-terminal S domain is membrane embedded and protrudes to the particle surface. These data suggest that some L molecules form a highly folded, potentially spring-loaded topology with five membrane-spanning regions and a membrane-traversing pre-S chain. PMID- 10666281 TI - Adeno-associated virus major Rep78 protein disrupts binding of TATA-binding protein to the p97 promoter of human papillomavirus type 16. AB - Adeno-associated virus type 2 (AAV) is known to inhibit the promoter activities of several oncogenes and viral genes, including the human papillomavirus type 16 (HPV-16) E6 and E7 transforming genes. However, the target elements of AAV on the long control region (LCR) upstream of E6 and E7 oncogenes are elusive. A chloramphenicol acetyltransferase assay was performed to study the effect of AAV on the transcription activity of the HPV-16 LCR in SiHa (HPV-positive) and C-33A (HPV-negative) cells. The results reveal that (i) AAV inhibited HPV-16 LCR activity in a dose-dependent manner, (ii) AAV-mediated inhibition did not require the HPV gene products, and (iii) the AAV replication gene product Rep78 was involved in the inhibition. Deletion mutation analyses of the HPV-16 LCR showed that regulatory elements outside the core promoter region of the LCR may not be direct targets of AAV-mediated inhibition. Further study with the electrophoretic mobility shift assay demonstrated that Rep78 interfered with the binding of TATA binding protein (TBP) to the TATA box of the p97 core promoter more significantly than it disrupted the preformed TBP-TATA complex. These data thus suggest that Rep78 may inhibit transcription initiation of the HPV-16 LCR by disrupting the interaction between TBP and the TATA box of the p97 core promoter. PMID- 10666282 TI - The c-myc locus is a common integration site in type B retrovirus-induced T-cell lymphomas. AB - Type B leukemogenic virus (TBLV) induces rapidly appearing T-cell leukemias. TBLV insertions near the c-myc gene were detectable in 2 of 30 tumors tested, whereas 80% of the tumors showed c-myc overexpression. TBLV insertions on chromosome 15 (including a newly identified locus, Pad7) may cause c-myc overexpression by cis acting effects at a distance. PMID- 10666283 TI - Therapeutic effect of anti-macrophage inflammatory protein 2 antibody on influenza virus-induced pneumonia in mice. AB - We investigated the effect of anti-macrophage inflammatory protein 2 immunoglobulin G (aMIP-2 IgG) on the progression of influenza virus-induced pneumonia in mice. When mice were infected with a mouse lung-adapted strain of influenza A/PR/8/34 virus by intranasal inoculation, neutrophil counts in the bronchoalveolar lavage fluid (BALF) increased in parallel with the kinetics of MIP-2 production, which peaked 2 days after infection. After intracutaneous injection of a dose of 10 or 100 microg of aMIP-2 IgG once a day on days 0 and 1, neutrophil counts in BALF on day 2 were reduced to 49 or 37%, respectively, of the value in the control infected mice administered anti-protein A IgG. The antibody administration also improved lung pathology without affecting virus replication. Furthermore, by prolonged administration with a higher or lower dose for up to 5 days, body weight loss became slower and finally 40% of mice in both treatment groups survived potentially lethal pneumonia. These findings suggest that MIP-2-mediated neutrophil infiltration during the early phase of infection might play an important role in lung pathology. Thus, MIP-2 was considered to be a novel target for intervention therapy in potentially lethal influenza virus pneumonia in mice. PMID- 10666284 TI - Sendai virus blocks alpha interferon signaling to signal transducers and activators of transcription. AB - We demonstrate here that Sendai virus (SeV) blocks alpha interferon (IFN-alpha) signaling to signal transducers and activators of transcription (STATs) in HeLa cells. IFN-alpha-stimulated tyrosine phosphorylation of STATs and subsequent formation of the IFN-stimulated gene factor 3 transcription complex were inhibited in SeV-infected cells, resulting in inefficient induction of IFN stimulated gene products. None of the components of the signaling pathway-type I IFN receptor subunits Jak1, Tyk2, Stat1, Stat2, and p48-was degraded. Moreover, tyrosine phosphorylation of Jak1 in response to IFN-alpha was unaffected at the early phase of infection, suggesting that oligomerization of the receptor subunits proceeded normally. In contrast to Jak1, IFN-alpha-stimulated tyrosine phosphorylation of Tyk2 was partially inhibited. Therefore, this partial inhibition of activation of Tyk2 probably contributes to the subsequent failure in the activation of STATs. PMID- 10666285 TI - Pseudotyping of glycoprotein D-deficient herpes simplex virus type 1 with vesicular stomatitis virus glycoprotein G enables mutant virus attachment and entry. AB - The use of herpes simplex virus (HSV) vectors for in vivo gene therapy will require the targeting of vector infection to specific cell types in certain in vivo applications. Because HSV glycoprotein D (gD) imparts a broad host range for viral infection through recognition of ubiquitous host cell receptors, vector targeting will require the manipulation of gD to provide new cell recognition specificities in a manner designed to preserve gD's essential role in virus entry. In this study, we have determined whether an entry-incompetent HSV mutant with deletions of all Us glycoproteins, including gD, can be complemented by a foreign attachment/entry protein with a different receptor-binding specificity, the vesicular stomatitis virus glycoprotein G (VSV-G). The results showed that transiently expressed VSV-G was incorporated into gD-deficient HSV envelopes and that the resulting pseudotyped virus formed plaques on gD-expressing VD60 cells, albeit at a 50-fold-reduced level compared to that of wild-type gD. This reduction may be related to differences in the entry pathways used by VSV and HSV or to the observed lower rate of incorporation of VSV-G into virus envelopes than that of gD. The rate of VSV-G incorporation was greatly improved by using recombinant molecules in which the transmembrane domain of HSV glycoprotein B or D was substituted for that of VSV-G, but these recombinant molecules failed to promote virus entry. These results show that foreign glycoproteins can be incorporated into the HSV envelope during replication and that gD can be dispensed with on the condition that a suitable attachment/entry function is provided. PMID- 10666286 TI - Effect of maternal nutrient restriction in early gestation on responses of the hypothalamic-pituitary-adrenal axis to acute isocapnic hypoxaemia in late gestation fetal sheep. AB - Epidemiological and experimental evidence suggests that maternal undernutrition during pregnancy may alter development of fetal organ systems. We have demonstrated previously that fetal hypothalamic-pituitary-adrenal (HPA) axis responses to exogenous corticotropin-releasing hormone (CRH) + arginine vasopressin (AVP), or adrenocorticotrophin hormone (ACTH), are reduced in fetuses of mildly undernourished ewes. To examine these effects further we tested HPA axis responses to acute isocapnic hypoxaemia in fetal sheep at 114-129 days gestation (dGA), following 15% reduction in maternal nutritional intake between 0 and 70 dGA. Fetuses from control (C) and nutrient-restricted (R) ewes were chronically catheterised and plasma ACTH and cortisol responses were determined at 114-115, 120-123 and 126-129 dGA during hypoxaemia (1 h) induced by lowering the maternal inspired O2 fraction (FI,O2). Basal plasma cortisol concentrations and HPA axis responses at 114-115 and 120-123 dGA did not differ between C and R fetuses. At 126-129 dGA, both plasma ACTH (P < 0.01) and cortisol (P < 0.05) responses were smaller in R fetuses compared to C fetuses. Fetal blood gas status, fetal body weight, body proportions and organ weights did not differ between the groups. We conclude that mild maternal undernutrition alters development of the fetal HPA axis producing a reduction in pituitary and adrenal responsiveness to endogenous stimuli. PMID- 10666287 TI - Management of peripheral arterial disease (PAD). TASC Working Group. TransAtlantic Inter-Society Consensus (TASC). PMID- 10666288 TI - The enzymatic formation of novel bile acid primary amides. AB - Bifunctional peptidylglycine alpha-amidating monooxygenase (PAM) catalyzes the copper-, ascorbate-, and O(2)-dependent cleavage of C-terminal glycine-extended peptides and N-acylglycines to the corresponding amides and glyoxylate. The alpha amidated peptides and the long-chain acylamides are hormones in humans and other mammals. Bile acid glycine conjugates are also substrates for PAM leading to the formation of bile acid amides. The (V(MAX)/K(m))(app) values for the bile acid glycine conjugates are comparable to other known PAM substrates. The highest (V(MAX)/K(m))(app) value, 3.1 +/- 0.12 x 10(5) M(-1) s(-1) for 3 sulfolithocholylglycine, is 6.7-fold higher than that for d-Tyr-Val-Gly, a representative peptide substrate. The time course for O(2) consumption and glyoxylate production indicates that bile acid glycine conjugate amidation is a two-step reaction. The bile acid glycine conjugate is first converted to an N bile acyl-alpha-hydroxyglycine intermediate which is ultimately dealkylated to the bile acid amide and glyoxylate. The enzymatically produced bile acid amides and the carbinolamide intermediates were characterized by mass spectrometry and two-dimensional (1)H-(13)C heteronuclear multiple quantum coherence NMR. PMID- 10666289 TI - Measurement of the main photooxidation products of 2'-deoxyguanosine using chromatographic methods coupled to mass spectrometry. AB - Analytical methods were developed for the measurement of the main photooxidation products of 2'-deoxyguanosine (dGuo), arising from either the type I (electron transfer) or the type II (singlet oxygen) photosensitization mechanism. Oxidation of dGuo by a type I mechanism leads to the predominant formation of 2, 2-diamino 5-[2-deoxy-beta-d-erythro-pentofuranosyl)amino]-5(2H)-oxazo lone. On the other hand, the two 4R and 4S diastereomers of 4-hydroxy-8-oxo-4,8-dihydro-2' deoxyguanosine are the main singlet oxygen-mediated dGuo oxidation products. The modified nucleosides were measured by either gas chromatography coupled to mass spectrometry after silylation of the sample or by high-performance liquid chromatography associated to tandem mass spectrometry. In order to improve the accuracy of the assays, isotopically labeled internal standards were synthesized for an isotope dilution mass spectrometry quantitation. The methods were successfully applied to the measurement of methylene blue- and riboflavin mediated 2'-deoxyguanosine photooxidation reactions. The advantages of the two above-mentioned methods are discussed on the basis of comparative sensitivity and accuracy. PMID- 10666290 TI - Cytochromes P450 2C1/2 and P450 2E1 are retained in the endoplasmic reticulum membrane by different mechanisms. AB - Cytochrome P450 (P450) 2C1/2 contains redundant endoplasmic reticulum (ER) retention signals and is excluded from the recycling pathway. Other P450s, such as P450 2E1, have been detected in the plasma membrane and Golgi apparatus. To examine whether the mechanisms of ER retention might differ for P450 2C1/2 and P450 2E1, chimeras of green flourescent protein and the full-length proteins, N terminal signal/anchor sequences, or the cytoplasmic catalytic domains from these proteins have been expressed in COS1 cells. Chimeras with either the N-terminal signal/anchor sequence or the cytoplasmic domain of P450 2C1/2 were retained in the ER and the distribution was not altered by treatment with nocodazole. A chimera with full-length P450 2E1 was located in the ER, but in contrast to P450 2C1/2, treatment with nocodazole resulted in redistribution to a vesicular pattern, which suggested that this protein was retained in the ER by a retrieval mechanism. In support of this possibility, the P450 2E1 chimera, but not the P450 2C1/2 chimera, was included in transport vesicles generated in an in vitro budding assay. A chimera with only the N-terminal signal/anchor sequence of P450 2E1 fused to green fluorescent protein was located in the ER and nocodazole treatment altered its distribution, whereas a chimera with only the cytoplasmic domain of P450 2E1 was not efficiently retained in the ER and accumulated primarily in the Golgi region. These results demonstrate that the mechanisms for retention in the ER of two closely related members of the P450 superfamily are different and that the N-terminal signal/anchor sequence contains the dominant retention signal. PMID- 10666291 TI - Copper-dependent formation of disulfide-linked dimer of S100B protein. AB - Previous cell biological studies demonstrated that S100B protein enhances neurite extension of cortical neurons and stimulates proliferation of glial cells. Although these activities of the protein are ascribed to its disulfide-linked dimeric form, there have been no indications as to how the dimer is formed in vivo. We have found by an in vitro study that it is produced by copper-dependent oxidation of noncovalent S100B dimer. The disulfide-linked dimer markedly stimulated nitric oxide production in a microglial cell line, BV2. Interestingly, the disulfide-linked dimer formation was found to be prevented by ascorbic acid. The copper-dependent formation of the dimer may not happen in vivo under normal conditions; however, under pathological conditions where copper is likely to be released from tissues and catalyze autoxidation of ascorbic acid, the dimer formation may proceed, resulting in the stimulated production of nitric oxide that would induce toxic signaling pathways. PMID- 10666292 TI - Ebselen induces apoptosis in HepG(2) cells through rapid depletion of intracellular thiols. AB - Ebselen, 2-phenyl-1,2-benzisoselenazol-3(2H)-one, is a synthetic seleno-organic compound with antioxidant capability. In the present study, we systematically examined the ability of ebselen to induce apoptosis in a human hepatoma cell line, HepG(2). Ebselen-induced apoptosis was evaluated by (i) TdT-mediated dUTP nick end labeling assay; (ii) analysis of sub-G1 cells; (iii) cell morphology, including cell size and granularity examination; and (iv) DNA gel electrophoresis. The results showed that ebselen was able to induce typical apoptosis in HepG(2) cells in a dose- and time-dependent manner. In order to explore the possible mechanisms involved in ebselen-induced apoptosis, the effect of ebselen on intracellular thiol concentrations including reduced glutathione (GSH) and protein thiols and the effect of N-acetylcysteine (NAC) and buthionine sulfoximine (BSO) pretreatment on ebselen-induced apoptosis were investigated. It was found that (i) ebselen rapidly depleted intracellular GSH and protein thiols, moreover, the depletion preceded the occurrence of apoptosis; (ii) NAC, a precursor of intracellular GSH synthesis, significantly alleviated ebselen induced apoptosis; and (iii) BSO, a specific inhibitor of intracellular GSH synthesis, augmented ebselen-induced apoptosis significantly. Taken together, the present study demonstrates that ebselen is able to induce apoptosis in HepG(2) cells, most probably through rapid depletion of intracellular thiols. PMID- 10666293 TI - Differential gene expression in the activation and maturation of human monocytes. AB - Differential-display or RNA fingerprint was applied to identify genes differentially expressed in monocyte maturation induced by an immunomodulating peptide on human peripheral blood mononuclear cells. Two unknown sequences (06c22 and 06c71) and p21 protein (cyclin dependent kinase inhibitor) were repressed, and three genes activated: Cathepsin D, DRP2 (dihydropirimidinase related protein 2), and gp91phox (91-kDa subunit of citochrome b(558)). Phenotype of evolving monocytes was analyzed by flow cytometry and mRNA level of identified genes determined by reverse transcription-PCR. The expression pattern of identified genes seemed to correlate with different monocyte subsets, monocyte-derived cells, and expected functional changes. After peptide addition, immature monocytes were initially activated, increasing the expression of CD25, CD69, and HLA-DR markers. This was accompanied by repression of p21 and the two unknown sequences, along with the simultaneous activation of Cathepsin D and DRP2. Later, the differentiation marker CD16 rose, and gp91phox gene expression activated. Further maturation led certain monocytes to express marker CD23 and gp91phox expression to reach a maximum, while Cathepsin D and DRP2 dropped to preactivation levels. Results reflect part of the evolution of immature monocytes toward macrophages and monocyte-derived dendritic cell precursors. PMID- 10666294 TI - Effects of histone deacetylase inhibitors on the Ah receptor gene promoter. AB - The aromatic hydrocarbon receptor (AhR) is a ligand-dependent basic helix-loop helix-PAS-containing transcription factor which is activated by chemicals such as 2,3,7,8-tetrachlorodibenzo-p-dioxin. Constitutive expression of the AhR gene occurs in a tissue- and developmentally specific manner and appears to be altered by chemicals which affect histone deacetylase (HDAC) activity in cells in culture. Here we have directly characterized the effects of two HDAC inhibitors, n-butyrate and trichostatin A, on the promoter activity of the murine AhR gene. HDAC inhibitors increased the constitutive activity of the AhR gene promoter in a luciferase reporter construct by five- to sevenfold in a dose- and time-dependent manner in several cell lines and was correlated with an increase in endogenous AhR activity in an AhR-deficient cell line. Deletion analysis of the upstream region of the AhR gene localized the HDAC inhibitor effect to a 167-bp region encompassing -77 to +90 of the AhR gene promoter. Cotransfection of an AhR promoter-luciferase reporter plasmid with a vector expressing the E1A(12s) oncoprotein, a negative regulator of p300, a protein with histone acetylase activity, decreased AhR promoter activity fivefold. Overall, our results support a role for histone acetylation in the transcriptional activity of the AhR gene promoter. PMID- 10666295 TI - The role of microtubules in the regulation of proteoglycan synthesis in chondrocytes under hydrostatic pressure. AB - Chondrocytes of the articular cartilage sense mechanical factors associated with joint loading, such as hydrostatic pressure, and maintain the homeostasis of the extracellular matrix by regulating the metabolism of proteoglycans (PGs) and collagens. Intermittent hydrostatic pressure stimulates, while continuous high hydrostatic pressure inhibits, the biosynthesis of PGs. High continuous hydrostatic pressure also changes the structure of cytoskeleton and Golgi complex in cultured chondrocytes. Using microtubule (MT)-affecting drugs nocodazole and taxol as tools we examined whether MTs are involved in the regulation of PG synthesis in pressurized primary chondrocyte monolayer cultures. Disruption of the microtubular array by nocodazole inhibited [(35)S]sulfate incorporation by 39 48%, while MT stabilization by taxol caused maximally a 17% inhibition. Continuous hydrostatic pressure further decreased the synthesis by 34-42% in nocodazole-treated cultures. This suggests that high pressure exerts its inhibitory effect through mechanisms independent of MTs. On the other hand, nocodazole and taxol both prevented the stimulation of PG synthesis by cyclic 0. 5 Hz, 5 MPa hydrostatic pressure. The drugs did not affect the structural and functional properties of the PGs, and none of the treatments significantly affected cell viability, as indicated by the high level of PG synthesis 24-48 h after the release of drugs and/or high hydrostatic pressure. Our data on two dimensional chondrocyte cultures indicate that inhibition of PG synthesis by continuous high hydrostatic pressure does not interfere with the MT-dependent vesicle traffic, while the stimulation of synthesis by cyclic pressure does not occur if the dynamic nature of MTs is disturbed by nocodazole. Similar phenomena may operate in cartilage matrix embedded chondrocytes. PMID- 10666296 TI - Isolation and characterization of a novel 530-kDa protein complex (PC530) capable of associating with the 20S proteasome from starfish oocytes. AB - A novel protein complex called PC530 was purified concomitantly with proteasomes from oocytes of the starfish, Asterina pectinifera, by chromatography with DEAE cellulose, phosphocellulose, Mono Q, and Superose 6 columns. The molecular mass of this complex was estimated to be 530 kDa by Ferguson plot analysis and about 500 kDa by Superose 6 gel filtration. Since the 1500-kDa proteasome fractions contain the PC530 subunits as well as the 20S proteasomal subunits, and also since the purified PC530 and the 20S proteasome were cross-linked with a bifunctional cross-linking reagent, it is thought that PC530 is able to associate with the 20S proteasome. The PC530 comprises six main subunits with molecular masses of 105, 70, 50, 34, 30, and 23 kDa. The 70-kDa subunit showed a sequence similarity to the S3/p58/Sun2/Rpn3p subunit of the 26S proteasome, whereas the other subunits showed little or no appreciable similarity to the mammalian and yeast regulatory subunits. These results indicate that starfish oocytes contain a novel 530-kDa protein complex capable of associating with the 20S proteasome, which is distinctly different from PA700 or the 19S regulatory complex in molecular size and subunit composition. PMID- 10666297 TI - Purification and characterization of NAD-dependent morphine 6-dehydrogenase from hamster liver cytosol, a new member of the aldo-keto reductase superfamily. AB - Morphine 6-dehydrogenase, which catalyzes the dehydrogenation of morphine to morphinone, was purified 815-fold to a homogeneous protein from the soluble fraction of hamster liver with a yield of 15%. The enzyme was a monomeric protein with a molecular weight of 38 kDa and an isoelectric point of 5.6. Although both NAD and NADP served as cofactors, the enzyme activity with NADP was less than 5% that found with NAD at pH 7.4. With NAD, the enzyme gave the maximal activity at pH 9.3, and the K(m) and V(max) values toward morphine were 1.0 mM and 0.43 unit/mg protein, respectively. Among morphine congeners, normorphine exhibited higher activity than morphine, but codeine and ethylmorphine were poor substrates, and dihydromorphine and dihydrocodeine showed no detectable activity. The enzyme also exhibited significant activity for a variety of cyclic and alicyclic alcohols. In addition to xenobiotics, the enzyme catalyzed the dehydrogenation of 17beta-hydroxysteroids with much higher affinities than morphine. In the reverse reaction, the enzyme exhibited high activity for o quinones, but morphinone, naloxone, and aromatic aldehydes and ketones were reduced at slow rates. Sulfhydryl reagents and ketamine strongly inhibited the enzyme, whereas pyrazole, barbital, and indomethacin had little effect on enzyme activity. 17beta-Hydroxysteroids inhibited the enzyme in a competitive manner against morphine. A total of 302 amino acid residues, which comprised approximately 94% of whole protein, were identified by sequencing of the peptides obtained by proteolytic digestion. This amino acid sequence of the enzyme showed significant homology to members of the aldo-keto reductase (AKR) superfamily and shared 63-64% identity with members of the AKR1C subfamily. These findings indicate that the enzyme is a new member of the AKR superfamily that is involved in steroid metabolism as 17beta-hydroxysteroid dehydrogenase as well as xenobiotic metabolism. PMID- 10666298 TI - Regulation in metabolic systems under homeostatic flux control. AB - The general properties of metabolic systems under homeostatic flux control are analyzed. It is shown that the main characteristic point for an enzyme in such a system is a sharp transition from limitation outside the system to limitation by some enzyme inside the system. A method for the quantitative treatment of the experimental dependence of metabolic flux on enzyme content is presented. The conception of "nonlimiting," "near-limiting," and "limiting" enzymes is developed for these systems. It is pointed out that reactions close to a thermodynamic equilibrium under normal conditions can considerably limit the homeostatic fluxes. The rules for regulation of fluxes in such systems are illustrated by the data obtained for transgenic plants with reduced activities of some Calvin-cycle enzymes and further examples. A comparison is made between the developed quantitative description of metabolic fluxes under homeostatic flux control and the methods of metabolic control analysis. PMID- 10666299 TI - Distribution of proteasomes and of the five proteolytic activities in rat tissues. AB - Five peptidase activities (ChT-L, T-L, PGPH, BrAAP, and SNAAP) of the proteasome, and its caseinolytic activity, were measured in crude extracts of 10 rat tissues under experimental conditions simulating those found in vivo, thereby eliminating the alterations observed with the purified enzyme. The total and individual peptidase activities varied considerably from one tissue to another, whereas the proteolytic activity measured with [(14)C]methylcasein varied no more than twofold. The tissue-specific variations in individual peptidase activities may reflect tissue-specific differences in proteasome subunit composition, or the presence of regulators. Immunological assay using an antibody directed against the iota (alpha1) subunit showed that there was no correlation between protein abundance and peptidase activity. The results also show that the different peptidase activities are not representative of proteasome distribution in the different tissues. PMID- 10666300 TI - Phospholipase A(2) is involved in thapsigargin-induced sodium influx in human lymphocytes. AB - Previously, we reported that emptying of intracellular Ca(2+) pools with endoplasmatic Ca(2+)-ATP-ase inhibitor thapsigargin leads to the Na(+) influx in human lymphocytes (M. Tepel et al., 1994, J. Biol. Chem. 269, 26239-26242). In the present study we examined the mechanism underlying the thapsigargin-induced Na(+) entry. We found that the thapsigargin-induced increase in Na(+) concentration was effectively inhibited by three structurally unrelated phospholipase A(2) (PLA(2)) inhibitors, p-bromophenacyl bromide, 3-(4-octadecyl) benzoylacrylic acid (OBAA), and bromoenol lactone (BEL). The thapsigargin-induced Na(+) influx could be mimicked by PLA(2) exogenously added to the lymphocyte suspension. In addition, thapsigargin stimulated formation of arachidonic acid (AA), the physiological PLA(2) product. AA induced Na(+) entry in a time- and concentration-dependent fashion. Both, thapsigargin-induced Na(+) influx and AA liberation were completely inhibited in the presence of tyrosine kinase inhibitor genistein but not in the absence of extracellular Ca(2+). Collectively, these data show that thapsigargin-induced Na(+) entry is associated with tyrosine kinase-dependent stimulation of PLA(2). PMID- 10666301 TI - Interaction of lactoferrin with ceruloplasmin. AB - When added to human blood serum, the iron-binding protein lactoferrin (LF) purified from breast milk interacts with ceruloplasmin (CP), a copper-containing oxidase. Selective binding of LF to CP is evidenced by the results of polyacrylamide gel electrophoresis, immunodiffusion, gel filtration, and affinity chromatography. The molar stoichiometry of CP:LF in the complex is 1:2. Near-uv circular dichroism spectra of the complex showed that neither of the two proteins undergoes major structural perturbations when interacting with its counterpart. K(d) for the CP/LF complex was estimated from Scatchard plot as 1.8 x 10(-6) M. The CP/LF complex is found in various fluids of the human body. Upon injection into rat of human LF, the latter is soon revealed within the CP/LF complex of the blood plasma, from where the human protein is substantially cleared within 5 h. PMID- 10666302 TI - Identification of a beta-like DNA polymerase activity in bovine heart mitochondria. AB - A new DNA polymerase activity, distinct from DNA polymerase gamma, has been identified in bovine heart mitochondria. First detected among proteins isolated in a complex with mitochondrial DNA, the DNA polymerase activity has been partially purified 47,000-fold. Enzyme activity separates from DNA polymerase gamma on several chromatographic columns and appears to copurify with a 38 +/- 2 kDa polypeptide. Unlike DNA polymerase gamma, this enzyme is relatively resistant to inhibition by N-ethylmaleimide and dideoxynucleotides, has moderately low monovalent and high divalent cation requirements, and possesses 20-fold-higher apparent K(m) values for deoxynucleotides. The enzyme polymerizes deoxynucleotides onto a primed template DNA in a relatively nonprocessive fashion and lacks a detectable 3' to 5' exonuclease activity. Many of these characteristics resemble a beta-like mitochondrial DNA polymerase previously identified in, and considered unique to, trypanosomes. We propose that the bovine and trypanosomal enzymes are related and represent a new class of ubiquitous mitochondrial DNA polymerases. PMID- 10666303 TI - Biophysical study of the perturbation of model membrane structure caused by seminal plasma protein PDC-109. AB - PDC-109, the major heparin-binding protein of bull seminal plasma, binds specifically to sperm choline lipids at ejaculation and mediates capacitation by stimulating cholesterol and phospholipid efflux. We carried out a biophysical study to investigate the membrane perturbation effect caused by PDC-109. Binding of PDC-109 to phosphatidylcholine model membranes was maximal at a 12:1 phosphatidylcholine to protein molar ratio. The process was independent of the membrane structure and involved a slight conformational change of the protein, compatible with an increased exposure to the solvent. PDC-109 binding to dimyristoylphosphatidylcholine prevented lipid molecules from participating in the gel-to-liquid phase transition, due to enhancement of both acyl chain disorder and interfacial hydration. Visualization of the lipid-protein complexes by electron microscopy showed surface irregularities and the presence of 10-nm particles. Permeability assays confirmed the PDC-109-induced disruption of the vesicles. This effect was not modified by heparin. However, presence of cholesterol inhibited the process in a concentration-dependent manner. PMID- 10666304 TI - The multiple active enzyme species of gamma-aminobutyric acid aminotransferase are not isozymes. AB - Purified gamma-aminobutyric acid aminotransferase (GABA-AT) from pig brain under certain conditions gave a single band on 12% NaDodSO(4)-PAGE, whereas two or three distinct bands were observed on 7.5% native PAGE. These multiple active species were isolated by 5% preparative gel electrophoresis and characterized by N-terminal sequencing and MALDI-TOF mass spectrometry. The results indicate that these active enzyme species are not GABA-AT isozymes in pig brain, but are the products of proteolysis of the N-terminus of GABA-AT, differing by 3, 7, and 12 residues from the full sequence (as deduced from the cDNA), respectively. Conditions for obtaining the nontruncated GABA-AT were found, and the potential cause for the proteolysis was determined. It was found that Na(2)EDTA inhibits the N-terminal cleavage during GABA-AT preparation from pig brain. The presence of Triton X-100 in the homogenization step is partially responsible for this proteolysis, and Mn(2+) strongly enhances the protease activity, suggesting the presence of a membrane-bound matrix metalloprotease that causes the N-terminal cleavage. PMID- 10666305 TI - Induction of arginase II in human Caco-2 tumor cells by cyclic AMP. AB - The objective of this study was to elucidate the mechanism by which cyclic AMP increases arginase activity in cultured human Caco-2 tumor cells. Caco-2 cells were incubated for 24 h in the presence of 8-bromo cyclic AMP or forskolin, and the cells were harvested, lysed, and assayed for total arginase activity. Both test agents increased arginase activity by twofold, and this was attributed to the induction of the arginase II isoform. Both arginase II mRNA and protein showed increased expression in response to 8-bromo cyclic AMP and forskolin, and these effects were inhibited by H-89 (protein kinase A inhibitor), enhanced by okadaic acid (phosphatase inhibitor), and enhanced by 1-methyl-3-isobutylxanthine (cyclic nucleotide phosphodiesterase inhibitor). Cyclic GMP did not appear to be involved in arginase II induction. These observations indicate that cyclic AMP stimulates arginase II gene expression by mechanisms involving activation of protein kinase A and consequent activation of appropriate transcription factors. PMID- 10666306 TI - Cloning and characterization of glyoxalase I from soybean. AB - Glyoxalase I and glutathione transferase (GST) are two glutathione-dependent enzymes which are enhanced in plants during cell division and in response to diverse stress treatments. In soybean, a further connection between these two enzymes has been suggested by a clone (Accession No. X68819) resembling a GST being described as a glyoxalase I. To characterize glyoxalase I in soybean, GmGlyox I resembling the dimeric enzyme from animals has been cloned from a cDNA library prepared from soybean suspension cultures. When expressed in Escherichia coli, GmGlyox I was found to be a 38-kDa dimer composed of 21-kDa subunits and unlike the enzyme from mammals showed activity in the absence of metal ions. GmGlyox I was active toward the hemithioacetal adducts formed by reacting methylglyoxal, or phenylglyoxal, with glutathione, homoglutathione, or gamma glutamylcysteine, showing no preference for homoglutathione adducts over glutathione adducts, even though homoglutathione is the dominant thiol in soybean. When the clone X68819 was expressed in E. coli, the respective recombinant enzyme was active as a GST rather than a glyoxalase and was termed GmGST 3. GmGST 3 was active as a homodimer (45 kDa) composed of 26-kDa subunits and showed a preference for glutathione over homoglutathione when conjugating 1 chloro-2,4-dinitrobenzene. Both enzymes are associated with cell division in soybean cultures, but GmGST 3 (0.4% total protein) was 40 times more abundant than GmGlyox I (0.01%). PMID- 10666307 TI - The importance of SRS-1 residues in catalytic specificity of human cytochrome P450 3A4. AB - The structural basis for the regioselective hydroxylation of Delta-4-3 ketosteroids by human CYP3A4 was investigated. Prior studies had suggested that the chemical reactivity of the allylic 6beta-position might have a greater influence than steric constraints by the enzyme. Six highly conserved CYP3A residues from substrate recognition site 1 were examined by site-directed mutagenesis. F102A and A117L showed no spectrally detectable P450. V101G and T103A exhibited a wild-type progesterone metabolite profile. Of five mutants at residue N104, only N104D yielded holoenzyme and exhibited the same steroid metabolite profile as wild-type. Of four mutants at position S119 (A, L, T, V), the three hydrophobic ones produced 2beta-OH rather than 6beta-OH progesterone or testosterone as the major metabolite. Kinetic analysis showed S(50) values similar to wild-type for S119A (progesterone) and S119V (testosterone), whereas the V(max) values for 2beta-hydroxysteroid formation were increased in both cases. All four mutants exhibited an altered product profile for 7-hexoxycoumarin side-chain hydroxylation, whereas the stimulation of steroid hydroxylation by alpha-naphthoflavone was similar to the wild-type. The results indicate that the highly conserved residue S119 is a key determinant of CYP3A4 specificity and reveal an important role of the active site topology in steroid 6beta hydroxylation. PMID- 10666308 TI - Carvedilol inhibits the exogenous NADH dehydrogenase in rat heart mitochondria. AB - There are several reports on the oxidation of external NADH by an exogenous NADH dehydrogenase in the outer leaflet of the inner membrane of rat heart mitochondria. Until now, however, little was known about its physiological role in cellular metabolism. The present work shows that carvedilol (?1-[carbazolyl (4)-oxy]-3-[2-methoxyphenoxyethyl)amino]-pro - panol-(2)?) is a specific inhibitor of an exogenous NADH dehydrogenase in rat heart mitochondria. Carvedilol does not affect oxygen consumption linked to the oxidation of succinate and internal NADH. It is also demonstrated that the inhibition of exogenous NADH dehydrogenase by carvedilol is accompanied by the inhibition of alkalinization of the external medium. In contrast to the addition of glutamate/malate or succinate, exogenous NADH does not generate a membrane potential in rat heart mitochondria, as observed with a TPP(+) electrode. It is also demonstrated that the oxygen consumption linked to NADH oxidation is not due to permeabilized mitochondria, but to actual oxidase activity in the inner membrane. The enzyme has a K(m) for NADH of 13 microM. Carvedilol is a noncompetitive inhibitor of this external NADH dehydrogenase with a K(i) of 15 microM. Carvedilol is the first inhibitor described to this organospecific enzyme. Since this enzyme was demonstrated to play a key role in the cardiotoxicity of anticancer drugs of the anthracycline family (e.g., adriamycin), we may suggest that the administration of carvedilol to tumor patients treated with adriamycin might be of great help in the prevention of the cardioselective toxicity of this antibiotic. PMID- 10666309 TI - Sterol regulatory element binding protein-mediated effect of fluvastatin on cytosolic 3-hydroxy-3-methylglutaryl-coenzyme A synthase transcription. AB - The effects of acute treatment with fluvastatin, a hypocholesteremic drug, on the mRNA levels of several regulatory enzymes of cholesterogenesis and of the LDL receptor were determined in rat liver. Fluvastatin increased the hepatic mRNA levels for HMG-CoA reductase up to 12-fold in 5 weeks of treatment at a daily dose of 6. 3 mg/kg. The effect was less marked in cytosolic HMG-CoA synthase, farnesyl-PP synthase, squalene synthetase, and LDL receptor. SREBP-2 mRNA levels were also increased, but SREBP-1 were not. De novo synthesis of cholesterol in several cultured cells was reduced by increasing concentrations of fluvastatin, and the IC(50) values of fluvastatin in HepG2, CV-1, and CHO cells were respectively 0.01, 0. 05, and 0.1 microM. When CHO cells stably transfected with a chimeric gene composed of the promoter of cytosolic HMG-CoA synthase and the CAT gene as a reporter were incubated with fluvastatin, the CAT gene was overexpressed, an effect which was similar to the cotransfection with the processed form of SREBP-1a. Both ALLN and fluvastatin increased the transcriptional activity of cytosolic HMG-CoA synthase. Mutation in either SRE or NF-Y boxes abolished the increase in transcriptional rate caused by fluvastatin in the promoter of cytosolic HMG-CoA synthase. These results indicate that the increase in transcriptional activity in the HMG-CoA synthase gene attributable to fluvastatin is a consequence of the activation of the proteolytic cleavage of SREBPs by reduced levels of intracellular cholesterol. PMID- 10666310 TI - Evidence for requirement of NADPH-cytochrome P450 oxidoreductase in the microsomal NADPH-sterol Delta7-reductase system. AB - Rabbit antibodies raised against the hydrophilic part of microsomal NADPH cytochrome P450 oxidoreductase (denoted fpT) demonstrated a marked ability to inhibit NADPH-sterol Delta7-reductase activity. In addition, trypsin and proteinase K treatment of microsomes removed almost all microsomal electron transfer constituents from the microsomes, but the Delta7-reductase activity could be reconstituted by adding detergent-solubilized NADPH-cytochrome P450 oxidoreductase (denoted OR). Furthermore, after solubilization from microsomes, the Delta7-reductase activity could be reconstituted with OR in a DEAE-cellulose column chromatography eluate fraction, which contained little OR activity. In the microsomal system, carbon monoxide, ketoconazole, and miconazole, specific inhibitors of cytochrome P450, had no effect on Delta7-reductase activity. These results provide the first evidence of an essential requirement of OR, which is distinct from cytochrome P450, in the NADPH-sterol Delta7-reductase system. EDTA, o-phenanthroline and KCN markedly lowered Delta7-reductase activity in a dose dependent manner. Among metal ions tested, only ferric ion restored the reductase activity in the EDTA-treated microsomes. These results sugguest that NADPH-sterol Delta7-reductase is membrane-bound iron-dependent protein embedded in the microsomal lipid bilayer. PMID- 10666311 TI - Effects of epinephrine infusion on expression of calpastatin in porcine cardiac and skeletal muscle. AB - beta-Adrenergic agonists induce muscle hypertrophy in mammalian species and alter the extractable activity of calpain proteinase and its specific endogenous inhibitor calpastatin. The effects on skeletal and cardiac muscle calpastatin of continuously infusing a group of pigs for 7 days with the physiological agonist epinephrine (0.15 microg/kg/min) were examined and compared with a placebo group. Basal levels of extractable calpastatin activity were higher in cardiac than skeletal muscle and epinephrine infusion increased the extractable activity in both muscle types (P < 0.05). An anti-recombinant porcine calpastatin antiserum detected a 135-kDa band and a 145/135-kDa doublet on Western blots of skeletal and cardiac extracts, respectively. Epinephrine infusion increased the 135-kDa band in skeletal muscle (P < 0.05), while the ratio of 145/135 kDa in cardiac muscle was decreased (P < 0.05). From Northern blots, the patterns of calpastatin mRNA species were similar in the two muscle types, two major transcripts at 5.8 and 3. 2 kb in cardiac muscle, with equivalent bands in skeletal muscle of 5.4 and 2.8 kb. A faint 7.9-kb band was also detected in skeletal muscle. Epinephrine infusion had no effect on skeletal calpastatin mRNA but tended to increase the 5.8-kb mRNA in cardiac muscle (P = 0. 053). These data indicate a differential response of the two muscle types to mildly elevated plasma epinephrine concentration and the response to elevated epinephrine may be at the translational or posttranslational level. Therefore, this type of stimulus appears to be less effective at perturbing calpastatin gene transcription than certain orally administered synthetic beta-adrenergic agonists. PMID- 10666312 TI - Ubiquitylation and destruction of endogenous c-mycS by the proteasome: are myc boxes dispensable? AB - c-MycS proteins are truncated forms of the transcription factor which have been shown to be produced by translation initiation at internal methionines (101, 121, and 134) and to be functional in the regulation of gene expression, cell proliferation, and apoptosis. Treatment of human leukemia HL60 cells with lactacystin, a specific inhibitor of the proteasome, increased the steady-state levels of endogenous c-MycS proteins. The half-life of endogenous [(35)S]MycS was similar to that of c-Myc ( approximately 23 min) in HL60 cells. c-Myc(Delta2 143), which lacks the transcription regulatory domain, had a half-life which was similar to that of endogenous c-Myc in 293 and HL60 cells. Treatment of the cells with lactacystin stabilized [(35)S]Myc(Delta2-143) and [(35)S]Myc and caused multi-ubiquitin conjugates of c-Myc, c-MycS, and Myc(Delta2-143) to accumulate. These findings indicate that the Myc homology boxes and the rest of the transcription regulatory domain (the first 144 amino acids) are dispensable for ubiquitylation and rapid destruction of c-MycS and c-Myc by the proteasome. PMID- 10666313 TI - TGF-beta1 stimulation of fibronectin transcription in cultured human lung fibroblasts requires active geranylgeranyl transferase I, phosphatidylcholine specific phospholipase C, protein kinase C-delta, and p38, but not erk1/erk2. AB - The cytokine transforming growth factor-beta (TGF-beta) has multiple effects on a variety of cell types, modulating cell growth and differentiation as well as extracellular matrix deposition and degradation. In the present work, we demonstrate that TGF-beta1 produces a fourfold increase in transcription of the fibronectin gene in cultured human fetal lung fibroblasts with only a small increase in mRNA stability resulting in a significant increase in fibronectin mRNA steady state level. A corresponding increase in production of fibronectin protein accompanied the increase in mRNA. Through the use of specific inhibitors, we demonstrate that geranylgeranylated, but not farnesylated or acylated protein(s), protein kinase C-delta, phosphatidylcholine-specific phospholipse C, tyrosine kinase activity, and stress-activated protein kinase p38 are required for this TGF-beta1 effect. Trimeric G proteins and mitogen-activated protein kinases erk1 and erk2 do not appear to be involved. While these results emphasize the complexities involved in the control of extracellular matrix synthesis by TGF beta, they also identify reaction sites that may be amenable to pharmacologic modulation. Such modulation could be of great advantage in the treatment of a wide variety of undesirable fibrotic reactions. PMID- 10666314 TI - Proteasome inhibition in neuronal cells induces a proinflammatory response manifested by upregulation of cyclooxygenase-2, its accumulation as ubiquitin conjugates, and production of the prostaglandin PGE(2). AB - Inclusions containing ubiquitin-protein aggregates appear in neurons of patients with neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease. The relationship between inclusion production and cell viability is not understood. To address this issue, we investigated the response of an established mouse neuronal cell line and of embryonic rat mesencephalic cultures to inhibition of the ubiquitin/proteasome pathway. Two proteasome inhibitors, a peptidyl aldehyde and an epoxy ketone, which cause accumulation of ubiquitinated proteins, were found to enhance expression of stress-inducible genes, including HSP70i and the polyubiquitin genes UbB and UbC. Under these conditions, mRNA and protein levels of the inducible form of cyclooxygenase (COX-2) were upregulated together with its product, PGE(2), a proinflammatory prostaglandin. Proteasomal inhibition also led to stabilization of COX-2 as ubiquitin conjugates, suggesting that the ubiquitin/proteasome pathway contributes to the regulation of COX-2 protein levels. Treatment with antioxidants known to inhibit NFkappaB and AP-1 transcriptional activation failed to abrogate COX-2 upregulation. Instead, these inhibitors exacerbated the stress response by potentiating HSP70i levels while eliciting a decrease in PGE(2) production. These findings suggest that the accumulation of ubiquitinated proteins resulting from proteasome inhibition in neuronal cells is associated with a proinflammatory response that may be an important contributor to neurodegeneration. PMID- 10666315 TI - Expression of 1,25-dihydroxyvitamin D(3) receptor in the immune system. AB - In addition to its role in calcium and skeletal homeostasis, there is increasing evidence that the hormonal form of vitamin D, 1, 25-dihydroxyvitamin D(3), appears to serve as a modulator of the immune system. We have determined the level of the 1, 25-dihydroxyvitamin D(3) receptor (VDR) in resting and activated lymphocytes by immuno- and ligand-binding assays. As expected from previous work, the total T lymphocyte population contains VDR whose levels are increased when activated and treated with 1, 25-dihydroxyvitamin D(3). Surprisingly, the highest concentrations of VDR are found in CD8 lymphocytes, although significant amounts are also present in CD4 lymphocytes. Furthermore, B lymphocytes do not contain detectable amounts of VDR. Cells of the monocyte/macrophage lineage possess small amounts of VDR that are not affected by activation but are increased by treatment with 1, 25-dihydroxyvitamin D(3). These results suggest that CD8 lymphocytes may be a major site of 1,25-dihydroxyvitamin D(3) action, while B lymphocytes are likely not directly regulated by 1, 25-dihydroxyvitamin D(3). PMID- 10666316 TI - Inhibition of c-Jun expression induces antioxidant enzymes under serum deprivation. AB - We previously reported that antisense c-jun suppressed apoptosis induced by serum deprivation in F-MEL cells. To elucidate the molecular mechanisms responsible for this suppression of apoptosis we investigated the activities and protein expression of antioxidant materials in the cell under serum deprivation. In the parental F-MEL cells enzyme activities of catalase, glutathione S-transferase (GST), and glutathione peroxidase (GPx) increased to reach the maximum at 24-72 h after removal of serum and then decreased to initial levels or a little less. Superoxide dismutase (SOD) maintained the initial level for 72 h and increased 1.5- to 2-fold at 96 h. Glutathione (GSH) levels increased at 24 h and then dropped significantly to one-third the initial level. On the other hand, in c junAS (+) cells, in which antisense c-jun was expressed and c-Jun protein expression was reduced to undetectable level. We found 1.9-, 2.7-, 4.8-, and 15. 8-fold increase in the activities of catalase, GST, SOD, and GPx, respectively, at 96 h. GSH maintained almost the same level as the initial. Enhancement of these enzyme activities in c-junAS (+) cells was induced under serum deprivation. Western blottings for catalase, GST, and SOD also showed enhanced increase in protein expression, supporting the increase in enzyme activities. Cellular peroxide level under serum deprivation was monitored by flow cytometry using DCFH DA as a probe. We found that the peroxide level increased at 24 h and then decreased at 72 and 96 h in c-junAS (+) cells, and reduction of the peroxide level coincided with an increase in antioxidant enzyme activities. These results indicate that antioxidant materials such as catalase, GST, SOD, GPx, and GSH are induced by serum deprivation when c-jun expression is inhibited in F-MEL cells. The link between inhibition of c-jun expression and enhancement of cellular antioxidant defense is discussed. PMID- 10666317 TI - Electron paramagnetic resonance studies of radical species of proanthocyanidins and gallate esters. AB - The polyphenols present in green tea or red wine comprise both regular flavon(ol)s and proanthocyanidins, i.e., derivatives of flavan-3-ols, whose distinct antioxidative potential is of great importance for explaining the beneficial effects of these nutrient beverages. Using EPR spectroscopy, we investigated radical structures obtained after oxidation of the parent compounds either by horseradish peroxidase/hydrogen peroxide or after autoxidation in moderately alkaline solutions. Both proanthocyanidins (monomers of condensed tannins, e.g., (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, (-) epigallocatechin, (-)-epigallocatechin gallate, Pycnogenol) and gallate esters (hydrolyzable tannins, e.g., propylgallate, beta-glucogallin, pentagalloyl glucose and tannic acid) yielded predominantly semiquinone structures derived from the parent catechol or pyrogallol moieties. Evidence for a time-dependent oligomerization was obtained for (-)-epigallocatechin gallate, supporting our hypothesis that o-quinones formed from the initial semiquinone disproportionation are prone to nucleophilic addition reactions. Such phenolic coupling reactions would retain the numbers of reactive catechol/pyrogallol structures and thus the antioxidative potential. We therefore propose that proanthocyanidins are superior antioxidants as compared to flavon(ol)s proper, whose quinones are more likely to redox-cycle and act as prooxidants. PMID- 10666318 TI - Distinct contributions of glycoprotein VI and alpha(2)beta(1) integrin to the induction of platelet protein tyrosine phosphorylation and aggregation. AB - Platelet activation by collagen depends principally on two receptors, alpha(2)beta(1) integrin (GPIa-IIa) and GPVI. During this activation, the nonreceptor protein tyrosine kinase pp72(syk) is rapidly phosphorylated, but the precise contribution of alpha(2)beta(1) integrin and GPVI to signaling for this phosphorylation is not clear. We have recently found that proteolysis of platelet alpha(2)beta(1) integrin by the snake venom metalloproteinase, jararhagin, results in inhibition of collagen-induced platelet aggregation and pp72(syk) phosphorylation. In order to verify whether the treatment of platelets with jararhagin had any effect on GPVI signaling, in this study we stimulated platelets treated with either jararhagin or anti-alpha(2)beta(1) antibody with two GPVI agonists, an antibody to GPVI and convulxin. Platelet shape change and phosphorylation of pp72(syk) by both GPVI agonists was preserved, as was the structure and function of GPVI shown by (125)I-labeled convulxin binding to immunoprecipitated GPVI from jararhagin-treated platelets. In contrast, defective platelet aggregation in response to GPVI agonists occurred in both jararhagin treated and alpha(2)beta(1)-blocked platelets. This apparent cosignaling role of alpha(2)beta(1) integrin for platelet aggregation suggests the possibility of a topographical association of this integrin with GPVI. We found that both platelet alpha(2)beta(1) integrin and GPVI coimmunoprecipitated with alpha(IIb)beta(3) integrin. Since platelet aggregation requires activation of alpha(IIb)beta(3) integrin, defective aggregation in the absence of alpha(2)beta(1) suggests that this receptor may provide a signaling link between GPVI and alpha(IIb)beta(3). Our study therefore demonstrates that platelet signaling leading to pp72(syk) phosphorylation initiated with GPVI engagement by either convulxin or GPVI antibody does not depend on alpha(2)beta(1) integrin. However, alpha(IIb)beta(3) integrin may, in this model, require functional alpha(2)beta(1) integrin for its activation. PMID- 10666319 TI - The Ang II-induced growth of vascular smooth muscle cells involves a phospholipase D-mediated signaling mechanism. AB - Angiotensin (Ang) II acts as a mitogen in vascular smooth muscle cells (VSMC) via the activation of multiple signaling cascades, including phospholipase C, tyrosine kinase, and mitogen-activated protein kinase pathways. However, increasing evidence supports signal-activated phospholipases A(2) and D (PLD) as additional mechanisms. Stimulation of PLD results in phosphatidic acid (PA) formation, and PA has been linked to cell growth. However, the direct involvement of PA or its metabolite diacylglycerol (DAG) in Ang II-induced growth is unclear. PLD activity was measured in cultured rat VSMC prelabeled with [(3)H]oleic acid, while the incorporation of [(3)H]thymidine was used to monitor growth. We have previously reported the Ang II-dependent, AT(1)-coupled stimulation of PLD and growth in VSMC. Here, we show that Ang II (100 nM) and exogenous PLD (0.1-100 units/mL; Streptomyces chromofuscus) stimulated thymidine incorporation (43-208% above control). PA (100 nM-1 microM) also increased thymidine incorporation to 135% of control. Propranolol (100 nM-10 microM), which inhibits PA phosphohydrolase, blocked the growth stimulated by Ang II, PLD, or PA by as much as 95%, an effect not shared by other beta-adrenergic antagonists. Propranolol also increased the production of PA in the presence of Ang II by 320% and reduced DAG and arachidonic acid (AA) accumulation. The DAG lipase inhibitor RHC-80267 (1 10 microM) increased Ang II-induced DAG production, while attenuating thymidine incorporation and release of AA. Thus, it appears that activation of PLD, formation of PA, conversion of PA to DAG, and metabolism of DAG comprise an important signaling cascade in Ang II-induced growth of VSMC. PMID- 10666320 TI - Molecular cloning of a taxa-4(20),11(12)-dien-5alpha-ol-O-acetyl transferase cDNA from Taxus and functional expression in Escherichia coli. AB - The taxa-4(20),11(12)-dien-5alpha-ol-O-acetyl transferase which catalyzes the third step of Taxol biosynthesis has been isolated from methyl jasmonate-induced Taxus cells, and partially purified and characterized (K. Walker, R. E. B. Ketchum, M. Hezari, D. Gatfield, M. Golenowski, A. Barthol, and R. Croteau, Arch. Biochem. Biophys. 364, 273-279 1999). A revised purification method allowed internal amino acid microsequencing of the enzyme, from which primers were designed and employed to amplify a transacetylase gene-specific fragment. This radiolabeled, 900-bp amplicon was used as a hybridization probe to screen a cDNA library constructed from poly(A)(+) RNA isolated from induced Taxus cells, from which a full-length transacetylase sequence was obtained. Expression of this clone from pCWori(+) in Escherichia coli JM109 cells yielded the functional enzyme, as determined by radiochemical assay and combined capillary gas chromatographic-mass spectrometric verification of the acetylated product. The full-length DNA has an open-reading frame of 1317 nucleotides corresponding to a deduced amino acid sequence of 439 residues that exhibits high sequence identity to the proteolytic fragments of the native enzyme, which the recombinant transacetylase resembles in properties. Consistent with the size of the operationally soluble native enzyme, the DNA appears to encode a monomeric protein of molecular weight 49,079 that bears no N-terminal organellar targeting information. Sequence comparison of the taxadien-5alpha-ol-O-acetyl transferase with the few other known acyl transferases of plant origin indicates a significant degree of similarity between these enzymes (64-67%). The efficient conversion of taxadien-5alpha-yl acetate to further hydroxylated intermediates of the Taxol pathway confirms the significance of this acylation step and suggests this taxadienol transacetylase to be an important target for genetic manipulation to improve Taxol production. PMID- 10666321 TI - Cloning, heterologous expression, and enzymological characterization of human squalene monooxygenase. AB - The cDNA for human squalene monooxygenase, a key enzyme in the committed pathway for cholesterol biosynthesis, was amplified from a human liver cDNA library and cloned, and the protein was expressed in Escherichia coli and purified. Kinetic analysis of the purified enzyme revealed an apparent K(m) for squalene of 7.7 microM and an apparent k(cat) of 1.1 min(-1). For FAD the apparent K(m) is 0.3 microM, consistent with a loosely bound flavin. The apparent K(m) for NADPH cytochrome P450 reductase, the requisite electron transfer partner, is 14 nM. The amount of reductase needed for maximal activity is about threefold less than the amount of squalene monooxygenase present in the assay; thus, electron transfer to the monooxygenase is not likely to be rate limiting. Previous reports have implicated inhibition of this enzyme as the cause of a peripheral demyelination seen in weanling rats fed a diet containing tellurium. As no data were available for humans, the ability of a number of tellurium and related elemental compounds to inhibit the recombinant human enzyme was examined. Tellurite, tellurium dioxide, selenite, and selenium dioxide were inhibitory; the tellurium compounds were more potent than the selenium compounds, as indicated by their IC(50) values (17 and 37 microM, respectively). Kinetic analysis of the inhibition by tellurite suggests multiple sites of interaction with the enzyme in a noncompetitive manner with respect to squalene. PMID- 10666322 TI - Lys-Ala mutations of type I adenylyl cyclase result in altered susceptibility to inhibition by adenine nucleoside 3'-polyphosphates. AB - Native and recombinant wild type and mutant forms of type I adenylyl cyclase, expressed in fall army worm ovarian cells (Sf9) cells, with mutations Lys-923 Ala, Lys-921-Ala, and Lys-350-Ala, retained the characteristic noncompetitive inhibition by adenine nucleoside 3'-polyphosphates, but exhibited substantially different sensitivities to inhibition by them. The type I K923A enzyme resulted in increased IC(50) values, e.g., >100-fold for 2'-deoxyadenosine-3' monophosphate, but the shift diminished as the number of 3'-phosphates increased. The K921A mutation increased IC(50) values approximately 5-fold for all adenine nucleosides tested, whereas the K350A mutation increased IC(50) values approximately 6- to 8-fold for all adenine nucleosides tested except 2' deoxyadenosine-3'-diphosphate, which was increased >/=2-fold. The data suggest that 3'-phosphates sufficiently increase binding affinity of these ligands to compensate for the reduced coordination of the adenine moiety induced by the K923A mutation. Moreover, the altered structures induced by both K350A and K921A mutations impair ligand binding in general, but paradoxically those resulting from the K350A change minimally affected nucleoside 3'-diphosphate binding, implying that selective changes in ligand binding can be induced by this site specific mutation. PMID- 10666323 TI - Relationship between total and free cellular Mg(2+) during metabolic stimulation of rat cardiac myocytes and perfused hearts. AB - The changes in total Mg were compared with changes in cytosolic free Mg(2+) during metabolic stimulation of collagenase-dispersed rat cardiac myocytes or Langendorff-perfused rat hearts. In myocytes the addition of agents leading to cAMP increase or protein kinase C activation results in a loss or gain of more than 5% of total Mg content within 3 min (i.e., 3-4 nmol Mg/mg protein). Under the same conditions, changes in cytosolic free Mg(2+) measured with fluorescent indicator are small and result in changes of cytosolic free Mg(2+) equivalent to 90-140 microM. In perfused hearts, beta-adrenergic stimulation results in a loss of total Mg larger than 0.5 micromol per gram of heart corresponding to 9% loss of total Mg content of the heart (estimated to be 5.8 micromol). Under these conditions there is no change in cytosolic free Mg(2+) or the major buffer of cytosolic Mg(2+), ATP, as measured by (31)P NMR. These data suggest that a major redistribution of total Mg occurs in intracellular organelles or in cytosolic buffers in order to maintain cytosolic free Mg(2+) relatively unchanged during the observed cellular massive translocation of total Mg. Hence, Mg(2+) may regulate metabolic functions not within the cytosol but in locations where its concentration oscillates, such as extracellular fluid and intracellular compartments. PMID- 10666324 TI - Gender differences in advanced mathematical problem solving. AB - Strategy flexibility in mathematical problem solving was investigated. In Studies 1 and 2, high school juniors and seniors solved Scholastic Assessment Test Mathematics (SAT-M) problems classified as conventional or unconventional. Algorithmic solution strategies were students' default choice for both types of problems across conditions that manipulated item format and solution time. Use of intuitive strategies on unconventional problems was evident only for high-ability students. Male students were more likely than female students to successfully match strategies to problem characteristics. In Study 3, a revised taxonomy of problems based on cognitive solution demands was predictive of gender differences on Graduate Record Examination-Quantitative (GRE-Q) items. Men outperformed women overall, but the difference was greater on items requiring spatial skills, shortcuts, or multiple solution paths than on problems requiring verbal skills or mastery of classroom-based content. Results suggest that strategy flexibility is a source of gender differences in mathematical ability assessed by SAT-M and GRE Q problem solving. PMID- 10666325 TI - A new model of verbal short-term memory. AB - Two experiments tested a neo-Piagetian model of verbal short-term memory and compared it with the articulatory loop model. Experiment 1 (n = 113, age range 9 11) tested word span for 2-, 3-, and 4-syllable words, with both visual and auditory presentation. Experiment 2 (with the same participants) tested recall of visually presented supraspan lists. Measures of M capacity (as conceived in Pascual-Leone's neo-Piagetian theory) and articulation rate were also used. The proposed model can account for the effects of M capacity, word length, and presentation modality. The fit of this model to the data was acceptable, and parameter estimates were consistent across experiments. Furthermore, a correlation was found between M capacity and word span which resisted partialling out of age and articulation rate. PMID- 10666326 TI - Movement substructures change as a function of practice in children and adults. AB - An experiment is reported in which participants at 6 (n = 20), 9 (n = 20), and 24 years (n = 20) of age either received or did not receive practice on a rapid aiming task using the arm and hand. The purpose of the experiment was to document the changes in movement substructures (in addition to movement time) as a function of practice. After receiving 10 baseline trials, subjects in the practice groups received 30 practice trials followed by 10 retention trials on each of 5 days, while subjects in the no-practice group had only baseline and retention trials. Retention-only trials were divided into primary (reflecting the ballistic controlled part of the movement) and secondary (reflecting corrective movement adjustments) submovements. In addition, jerk (the 3rd derivative of movement displacement) was calculated as an estimate of the smoothness of the movement. Participants increased the primary submovement as a function of practice; however, the increases were substantially larger in the children (25 30%) than in the adults (10%). Participants also decreased jerk as a function of practice and the decreases were greater in children than in adults. The results suggest that with practice the primary submovement is lengthened so that it ends nearer the target, especially in children. Associated with the primary submovement covering a larger percentage of the movement length and time, movements became smoother. PMID- 10666327 TI - Fibrous structures within the matrix of developing chick embryo mitochondria. PMID- 10666328 TI - Vertebrate mitochondrial DNA-a circle of surprises. AB - Evidence for the existence of a vertebrate mitochondrial genome first arose over 30 years ago. Application of emerging techniques of molecular biology established the structure of vertebrate mitochondrial DNA (mtDNA) as a small closed-circular species. The ability to purify these mtDNAs to a high degree facilitated studies on the overall replication and expression pattern of the genome. With the acquisition of the genomic sequences of human and mouse mtDNAs, it was possible to infer the genetic organization and some of the genes contained therein, as well as providing a basis for developing strategies to assign important regulatory elements involved in mtDNA replication and transcription. This, in turn, presented the opportunity to identify nucleus-encoded proteins that target to mtDNA and, in doing so, determine the replication and expression modes of the genome. Vertebrate cells, in general, need mtDNA due to the requirements for maintaining a functional oxidative phosphorylation pathway. Depression of mtDNA content or mutations in mtDNA can result in metabolic dysfunction severe enough, in some cases, to result in human lethality. The emergence of mouse models for human mitochondrial diseases should provide the experimental context to understand the full role of mtDNA in different cells, tissues, and organs; the control of organelle biogenesis; and the development of therapeutic strategies for treatment of mitochondrial disorders. PMID- 10666329 TI - Integrins alpha5beta1, alphavbeta1, and alphavbeta6 collaborate in squamous carcinoma cell spreading and migration on fibronectin. AB - The expression of alphavbeta6 fibronectin/tenascin receptor integrin is induced in malignant transformation of oral epithelium. In this study, we demonstrate the contribution of alphavbeta6 as well as other fibronectin receptor integrins in squamous cell carcinoma (SCC) cell adhesion and migration. Of 11 SCC cell lines isolated from the head and neck area, 8 (73%) expressed alphavbeta6 integrin on the cell surface. Three cell lines were chosen for further functional experiments: 1 with relatively high, 1 with moderate, and 1 with minimal surface expression of alphavbeta6 integrin. In addition to alphavbeta6, all 3 cell lines expressed alpha5beta1 and alphavbeta1 fibronectin receptor integrins. Function blocking experiments with inhibitory anti-integrin antibodies showed that all these three integrins were functional in SCC cell spreading on fibronectin. Integrin alphavbeta6, however, was not used as a primary but as an alternative fibronectin receptor by SCC cells, as the inhibitory anti-beta6 integrin antibody alone had no effect on spreading. In migration, however, alphavbeta6, alpha5beta1, and alphavbeta1 integrins were all used in cooperation. The presence of alphavbeta1 integrin in SCC cells is a novel finding as is its contribution to SCC cell migration. When one or two of these three receptors were blocked, the cells demonstrated an adaptive ability to remain migratory using integrins that were not targeted by antibodies. Utilization of a combination of receptors of different affinities may be beneficial for SCC cell migration versatility. PMID- 10666330 TI - Ceramide-induced apoptosis in fas-resistant Hodgkin's disease cell lines is caspase independent. AB - We investigated whether cell-permeable, synthetic ceramide (C6 ceramide) could induce apoptosis in Fas-resistant Hodgkin's disease (HD)-derived cell lines. Despite strongly expressing the Fas-receptor, two of three HD-derived cell lines were resistant to Fas-mediated apoptosis. This resistance to Fas could not be attributed to differential Fas isoform generation patterns between the Fas resistant and the Fas-sensitive cell lines. The Fas-resistant cell lines did not demonstrate the presence of Fas exon 8 deletion. Bcl-2 and BclxL levels were comparable between the Fas-resistant and the Fas-sensitive cell lines. C6 ceramide could induce apoptosis in both Fas-resistant cell lines and this was associated with a decrease in BclxL level. Caspase-1, caspase-3, or pan-caspase inhibitors could not prevent ceramide-induced apoptosis. Furthur, ceramide treatment did not lead to cleavage of caspase 3 or poly(ADP-ribose) polymerase, but caused a loss in mitochondrial transmembrane potential which could not be prevented by caspase inhibitors. Thus, we conclude that ceramide-induced apoptosis in Fas-resistant HD cell lines is caspase independent. PMID- 10666331 TI - Detection of apoptosis induced by topoisomerase inhibitors and serum deprivation in syrian hamster embryo cells. AB - The sensitivity of normal diploid Syrian hamster embryo (SHE) cells to apoptosis was tested after treatment with the topoisomerase inhibitors camptothecin and etoposide and after serum withdrawal. Programmed cell death (PCD) was identified through morphological, biochemical, and molecular changes and compared with that of HL60 cell line. The results showed that topoisomerase inhibitors, which were shown to be potent PCD inducers in the HL60 cell line, induced a weaker apoptotic response in SHE cells than after growth factor deprivation. In addition, serum free medium, which rapidly induced apoptosis in SHE cells, did not affect the HL60 cell line. In both cell types, PCD was expressed by condensed chromatin, fragmented nuclei, and DNA laddering on electrophoretic gels, an indisputable sign of apoptosis. In apoptotic HL60 cells, the cleavage of 113-kDa poly(ADP ribose)polymerase (PARP) resulted in the so-called apoptotic 89-kDa fragment and was associated with increased caspase-3 activity. In apoptotic SHE cells, PARP degraded early but the degradation profile was not characterized by the appearance of an 89-kDa fragment. Moreover, no activation of caspase-3 was noted. ZnCl(2), which is known to prevent protease activity responsible for apoptosis features, inhibited PARP cleavage and nuclear modifications induced by apoptotic stimuli in both cell types, but with a higher sensitivity in SHE cells. Apoptosis induced by serum deprivation was linked with c-myc negative regulation in SHE cells, but not with p53 protein accumulation, while topoisomerase inhibitors led to p53 stabilization without any change in c-myc expression. Serum-free medium and topoisomerase inhibitors did not modify c-myc expression in the HL60 cell line. The overall results demonstrated that apoptosis, which is a carefully regulated process of cell death, may proceed through mechanisms varying according to cell type or apoptosis inducer. In addition, markers which are generally considered hallmarks of apoptosis may fail to appear in some cell types. PMID- 10666332 TI - Gap-junctional coupling measured by flow cytometry. AB - A method is described which reliably quantifies the degree of intercellular communication via gap junctions by combining a dye-loading technique with fluorescence-activated flow cytometry. Our experiments expand former measurements of other groups by analyzing the time- and density-dependent onset of coupling with a fixed ratio of donor to recipient cells. The high sensitivity of this technique provides a better resolution than the microelectrode technique and allows the detection of small changes in gap-junctional coupling by examining a large number of cells in a single experiment. Suspended cells were loaded with the membrane-permeable dye calcein AM, which is intracellularly hydrolyzed by nonspecific esterases, and the resulting polyanionic calcein is thus trapped inside these donor cells. Gap junctions, however, are permeable for this fluorescent dye, as can be observed when suspended donor cells are added to recipient cells (i.e., monolayer cultures) in which case cell-cell contact is established within less than 60 min. In addition, one of these two cell populations can also be stained with a membrane-resident dye (e.g., DiI), which facilitates the identification of different cell populations (donors, recipients, and noncoupled cells) not only by epifluorescence microscopy but also by flow cytometry. Our analyses reveal that junctional coupling depends not only on the connexin type (homo- or heterotypic junction) but also on the origin (species) of the contacting cells (homo- or heterospecific contact). We confirm earlier reports in which homotypic-homospecific coupling was demonstrated with different techniques in connexin-transfected HeLa and RIN cells as well as in BICR/M1R(k) and 3T3/SV40 cells. In contrast to other publications, we show that a significant heterotypic-homospecific coupling between Cx40- and Cx43-HeLa transfectants can be resolved, whereas no coupling was detected for heterotypic-heterospecific contacts between Cx40-HeLa transfectants and the Cx43-expressing cell lines BICR/M1R(k), 3T3/SV40, and RIN. PMID- 10666333 TI - Expression of the adenovirus receptor and its interaction with the fiber knob. AB - The coxsackievirus group B (CVB) and adenovirus (Ad) receptor (HCVADR, formerly HCAR) is a cell surface protein with two immunoglobulin-like regions (IG1 and IG2) that serves as a receptor for two structurally unrelated viruses. We have established the tissue distribution of the receptor in the rodent by immunohistochemistry and show that the receptor is broadly expressed during embryonic development in the central and peripheral nervous systems and in several types of epithelial cells. The tissue distribution is more restricted in the adult but remains high mainly in epithelial cells. Using site-directed mutagenesis, based on computer modeling of the IG1 region, Ad5 binding could be inhibited but CVB attachment was unaffected. A double amino acid substitution in a three-stranded anti-parallel beta sheet that may form a face of the receptor completely inhibited Ad5 binding. Therefore, we conclude that the molecular interactions critical for Ad5 binding to HCVADR do not overlap with those of CVB3. In fact a specific antibody interfering with only CVB binding recognizes the IG2 domain in the receptor, suggesting that the CVB interacts with this region or an overlap between the IG1 and the IG2 regions. PMID- 10666334 TI - NPC1-containing compartment of human granulosa-lutein cells: a role in the intracellular trafficking of cholesterol supporting steroidogenesis. AB - Steroidogenic cells represent unique systems for the exploration of intracellular cholesterol trafficking. We employed cytochemical and biochemical methods to explore the expression, regulation, and function of the Niemann-Pick C1 protein (NPC1) in human granulosa-lutein cells. NPC1 was localized in a subset of lysosome-associated membrane glycoprotein 2 (LAMP-2)-positive vesicles. By analyzing the sensitivity of NPC1 N-linked oligosaccharide chains to glycosidases and neuraminidase, evidence was obtained for movement of nascent NPC1 from the endoplasmic reticulum through the medial and trans compartments of the Golgi apparatus prior to its appearance in cytoplasmic vesicles. NPC1 protein content and the morphology and cellular distribution of NPC1-containing vesicles were not affected by treatment of the granulosa-lutein cells with 8-Br-cAMP, which stimulates cholesterol metabolism into progesterone. In contrast, steroidogenic acute regulatory (StAR) protein levels were increased by 8-Br-cAMP. Incubation of granulosa-lutein cells with low-density lipoprotein (LDL) in the presence of the hydrophobic amine, U18666A, caused accumulation of free cholesterol in granules, identified by filipin staining, that contained LAMP-2 and NPC1. These granules also stained for neutral lipid with Nile red, reflecting accumulation of LDL derived cholesterol esters. LDL-stimulated progesterone synthesis was completely blocked by U18666A, leaving steroid output at levels similar to those of cells incubated in the absence of LDL. The hydrophobic amine also blocked the LDL augmentation of 8-Br-cAMP-stimulated progesterone synthesis, reducing steroid production to levels seen in cells stimulated with 8-Br-cAMP in the absence of LDL. Steroidogenesis recovered after U18666A was removed from the culture medium. U18666A treatment caused a 2-fold or more increase in NPC1 protein and mRNA levels, suggesting that disruption of NPC1's function activates a compensatory mechanism resulting in increased NPC1 synthesis. We conclude that the NPC1 compartment plays an important role in the trafficking of LDL-derived substrate in steroidogenic cells; that NPC1 expression is up-regulated when NPC1 action is blocked; and that the NPC1 compartment can be functionally separated from other intracellular pathways contributing substrate for steroidogenesis. PMID- 10666335 TI - Surfactant protein A regulates pulmonary surfactant secretion via activation of phosphatidylinositol 3-kinase in type II alveolar cells. AB - Pulmonary surfactant is secreted by the type II alveolar cells of the lung, and this secretion is induced by secretagogues of several types (e.g., ionomycin, phorbol esters, and terbutaline). Secretagogue-induced secretion is inhibited by surfactant-associated protein A (SP-A), which binds to a specific receptor (SPAR) on the surface of type II cells. The mechanism of SP-A-activated SPAR signaling is completely unknown. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 rescued surfactant secretion from inhibition by SP-A. In order to directly demonstrate a role for PI3K in SPAR signaling, PI3K activity was immunoprecipitated from type II cell extracts. PI3K activity increased rapidly after SP-A addition to type II cells. Since many receptors that activate PI3K do so through tyrosine-specific protein phosphorylation, antisera to phosphotyrosine, insulin-receptor substrate-1 (IRS-1), or SPAR were also examined. These antisera coimmunoprecipitated PI3K activity that was stimulated by SP-A. In addition, the tyrosine-specific protein kinase inhibitors genistein and herbimycin A blocked the action of SP-A on surfactant secretion. We conclude that SP-A signals to regulate surfactant secretion through SPAR, via pathways that involve tyrosine phosphorylation, include IRS-1, and entail activation of PI3K. This activation leads to inhibition of secretagogue-induced secretion of pulmonary surfactant. PMID- 10666336 TI - Increased TGFbeta type II receptor expression suppresses the malignant phenotype and induces differentiation of human neuroblastoma cells. AB - TGFbeta can modulate neuroblastoma (NB) cell proliferation and differentiation in vitro. In this study we used a NB cell line (LAN-5) which has been shown to partially respond to TGFbeta and to present high levels of TGFbeta receptor type I and low levels of receptor type II (TbetaRII) on the cell surface. To evaluate the role of TbetaRII in mediating TGFbeta effects, LAN-5 cells were transfected with an expression vector containing the human full-length TbetaRII cDNA or with the empty vector pcDNA3. Compared to control CLV3 cells (transfected with empty plasmid) and parental LAN-5 cells, isolated neomycin-resistant clones (CL1 and CL3) expressed higher levels of TbetaRII, had reduced cell growth rate in vitro, and were unable to form tumors in vivo. Furthermore, isolated clones modified their morphology, assuming a terminally differentiated neuronal phenotype. Immunocytochemical staining demonstrated a basal increased expression of neural specific markers, such as axonal growth-associated protein (GAP43) and neurofilaments (NF200). TGFbeta treatment further increased the synthesis of NF200 and GAP43 in the transfected clones as revealed by Western blot analysis. These data indicate that TbetaRII overexpression potentiates the TGFbeta signal transduction pathway, reverting NB cell neoplastic phenotype with the reduction of proliferation rate and the induction of terminal maturation. PMID- 10666338 TI - Telomere shortening by cisplatin in yeast nucleotide excision repair mutant. AB - Telomeres are unique DNA tandem repeats that form the ends of eukaryotic chromosomes to protect the chromosomes from degradation and illegitimate recombination. In yeast, loss of telomere may be compensated for through the acquisition of new telomere by RAD52-mediated or RAD52-independent recombinational repair. In this report, the effects of cis-dichlorodiammine platinum (II) (cisplatin) on telomere length and the role of nucleotide excision repair in telomere maintenance were examined in the yeast Saccharomyces cerevisiae. We showed that the SSL2 (RAD25) DNA repair yeast mutant exhibited a gradual shortening of the telomere in the presence of cisplatin. Further telomere shortening was prevented upon the withdrawal of cisplatin. Complementation of the mutant with the wild-type SSL2 (RAD25) gene abolished the cisplatin-induced telomere degradation. These results suggest that telomeres are susceptible to cisplatin-induced intrastrand crosslinks and that Ssl2 (Rad25) or the nucleotide excision repair pathway may play a critical role in the repair and the maintenance of telomere integrity. PMID- 10666337 TI - Human topoisomerase IIalpha and IIbeta interact with the C-terminal region of p53. AB - The p53 tumor suppressor protein is a critical regulator of cell cycle progression and apoptosis following exposure of cells to DNA damaging agents such as ionizing radiation or anticancer drugs. An important group of anticancer drugs, including compounds such as etoposide and doxorubicin (Adriamycin), interacts with DNA topoisomerase II (topo II), causing the accumulation of enzyme DNA adducts that ultimately lead to double-strand breaks and cell death via apoptosis. Human topo IIbeta has previously been shown to interact with p53, and we have extended this analysis to show that both topo IIalpha and IIbeta interact with p53 in vivo and in vitro. Furthermore, we show that the regulatory C terminal basic region of p53 (residues 364-393) is necessary and sufficient for interaction with DNA topo II. PMID- 10666339 TI - The carboxy terminus of AFAP-110 modulates direct interactions with actin filaments and regulates its ability to alter actin filament integrity and induce lamellipodia formation. AB - The actin filament-associated protein AFAP-110 is an SH2/SH3 binding partner for Src. AFAP-110 contains several protein-binding motifs in its amino terminus and has been hypothesized to function as an adaptor molecule that could link signaling proteins to actin filaments. Recent studies using deletional mutagenesis demonstrated that AFAP-110 can alter actin filament integrity in SV40 transformed Cos-1 cells. Thus, AFAP-110 may be positioned to modulate the effects of Src upon actin filaments. In this report, we sought to determine whether (a) AFAP-110 could interact with actin filaments directly and (b) deletion mutants could affect actin filament integrity and cell shape in untransformed fibroblast cells. The data demonstrate that the carboxy terminus of AFAP-110 is both necessary and sufficient for actin filament association, in vivo and in vitro. Analysis of the carboxy terminus revealed a mean 40% similarity with other known actin-binding motifs, indicating a mechanism for binding to actin filaments. AFAP 110 can also induce lamellipodia formation. Contiguous with the alpha-helical, actin-binding motif is an alpha-helical, leucine zipper motif. Deletion of the leucine zipper motif (AFAP(Deltalzip)) followed by cellular expression enabled AFAP(Deltalzip) to alter actin filament integrity and cell shape in untransformed cells as evidenced by the induction of lamellipodia formation. We hypothesize that AFAP-110 may be an important signaling protein that can directly modulate changes in actin filament integrity and induce lamellipodia formation. PMID- 10666340 TI - Spermatogenesis in mice is not affected by histone H1.1 deficiency. AB - The linker histone subtype H1.1 belongs to the group of main-type histones and is synthesized in somatic tissues as well as in germ cells during the S phase of the cell cycle. In adult mice the histone gene H1.1 is expressed mainly in thymus, spleen, and testis. The single-copy gene coding for the H1.1 protein was eliminated by homologous recombination in mouse embryonic stem cells. Mice homozygous for the deficient H1.1 gene developed normally until the adult stage without H1.1 mRNA and H1.1 protein. No anatomic abnormalities could be detected. In addition, mice lacking the H1.1 gene were fertile and they showed normal spermatogenesis and testicular morphology. PMID- 10666341 TI - Cleavage of Poly(ADP-ribose) polymerase measured in situ in individual cells: relationship to DNA fragmentation and cell cycle position during apoptosis. AB - Poly(ADP-ribose) polymerase (PARP), a nuclear enzyme involved in DNA repair, is a target of caspases during apoptosis: its cleavage onto 89- and 24-kDa fragments is considered to be a hallmark of the apoptotic mode of cell death. Another hallmark is the activation of endonuclease which targets internucleosomal DNA. The aim of the present study was to reveal cell cycle phase specificity as well as the temporal and sequence relationships of PARP cleavage vis-a-vis DNA fragmentation in two model systems of apoptosis, one induced by DNA damage via cell treatment with camptothecin (CPT) (mitochondria-induced pathway) and another by the cytotoxic ligand tumor necrosis factor alpha (TNF-alpha) (cell surface death receptor pathway). PARP cleavage was detected immunocytochemically using antibody which recognizes its 89-kDa fragment (PARP p89) while DNA fragmentation was assayed by in situ labeling of DNA strand breaks. The frequency and extent of PARP cleavage as well as DNA fragmentation were measured by mutiparameter flow and laser scanning cytometry. PARP cleavage, selective to S phase cells, was detected 90 min after administration of CPT. PARP cleavage in the cells treated with TNF-alpha was not selective to any cell cycle phase and was seen already after 30 min. DNA fragmentation trailed PARP cleavage by about 30 min and showed a similar pattern of cell cycle specificity. PARP p89 was present in nuclear chromatin but at least in the early phase of apoptosis it did not colocalize with DNA strand breaks. The rate of cleavage of PARP molecules in individual cells whether induced by CPT or TNF-alpha was rapid as reflected by the paucity of cells with a mixture of cleaved and noncleaved PARP molecules. In contrast, DNA fragmentation proceeded stepwise before reaching the maximal number of DNA strand breaks. Although time windows for PARP cleavage vs DNA fragmentation were different at early stages of apoptosis, a good overall correlation between the cytometric assays of apoptotic cells identification based on these events was observed in both CPT- and TNF-alpha-treated cultures. PMID- 10666342 TI - International differences: selection, noise, or real? PMID- 10666343 TI - Aortic stiffness: a predictor of acute coronary events? PMID- 10666344 TI - Early risk-stratification in patients with angina but non-diagnostic ECG. PMID- 10666345 TI - Cerebral hypoperfusion and impaired cerebral function in cardiac failure. PMID- 10666346 TI - Triage of chest pain patients based on early risk stratification using sensitive cardiac markers. PMID- 10666347 TI - Assessment of coronary heart disease risk in populations with different levels of risk. PMID- 10666348 TI - Athletic left ventricular hypertrophy: long-term studies are required. PMID- 10666349 TI - Beta-blockers continue to surprise us. PMID- 10666350 TI - Comparison of the Framingham risk function-based coronary chart with risk function from an Italian population study. AB - AIMS: The aim is to compare the coronary risk chart published by the European Task Force for Prevention of Coronary Heart Disease and produced using a Framingham risk function, with a risk function derived from an Italian population study. METHODS AND RESULTS: Coronary risk function in this study was the result of longitudinal experience in an Italian middle-aged population of 1656 male subjects followed-up for 25 years. To comply with the Framingham equation the same risk factors (age, systolic blood pressure, total serum cholesterol and smoking habits), end-points (any possible coronary event including angina pectoris), and length of follow-up (10 years) were used, and the model (log linear accelerated time failure model, accommodating the Weibull distribution) was similar. Comparisons were made computing the coronary risk for each cell of the coronary risk chart for men aged 40, 50 and 60 years. The Italian risk function produced highly significant coefficients for all four risk factors. Forty-four out of a total of 120 cells had a coronary risk of 20% or more in 10 years following the coronary risk chart, whereas this was reduced to four while using the Italian risk function (P<0.001). The Italian risk function largely underestimated the corresponding levels produced by the coronary risk chart and vice versa. CONCLUSION: The Framingham risk function-based coronary risk chart overestimates absolute coronary risk in countries characterized by a lower incidence of coronary events and should be used with caution. PMID- 10666351 TI - Geographic variability in outcomes within an international trial of glycoprotein IIb/IIIa inhibition in patients with acute coronary syndromes. Results from PURSUIT. AB - AIMS: Variations in outcome of patients from different geographic regions have been observed in many large international trials. We analysed the factors that might contribute to the geographic variations in patient outcome and treatment effect as observed in the PURSUIT trial. METHODS: In PURSUIT, 9461 patients with acute coronary syndromes without persistent ST-elevation were randomized to the platelet inhibitor eptifibatide or placebo for 72 h in 27 countries in four geographic regions: Western (n=3697) and Eastern Europe (n=1541) as well as North (n=3827) and Latin America (n=396). The primary end-point was the 30-day composite of death or myocardial infarction. In the initial univariate analysis, the treatment effect appeared greater in N. America than in W. Europe, while no benefit was apparent in L. America and E. Europe. However, the confidence intervals were wide and overlapping. To study these differences, a subdivision in an early and late patient outcome and treatment effect was made. Accordingly, we analysed the rate of death or infarction at 72 h censored for percutaneous coronary intervention and the rate between 3 and 30 days, respectively. Additional analyses were performed with different definitions of myocardial infarction using progressively higher thresholds of CK(-MB) elevation. Multivariable analysis was used to evaluate the relation between region and outcome and to determine the adjusted odds ratios for the eptifibatide treatment effect. RESULTS: Major differences in baseline demographics were apparent among the four regions; in particular, more patients from E. Europe had characteristics associated with impaired outcome. Interventional treatment also varied considerably, with more patients from N. America undergoing revascularization. Despite differences in the 72 h event rate, eptifibatide showed a consistent trend towards a reduction in the composite end-point among all four regions and for all definitions of infarction. Relative reductions ranged from 17-42% in W. Europe, 23-35% in N. America, 0-33% in E. Europe, and 55-82% in L. America. After multivariable adjustment, the pattern of benefit with eptifibatide was consistent among the regions. In patients undergoing percutaneous coronary intervention during study drug infusion in W. Europe (n=266) and N. America (n=931), the relative reduction in myocardial infarction during medical therapy ranged from 56 75% in W. Europe and 14-67% in N. America, while the reduction in procedure related events ranged from 12-44% and 25-61% for different definitions of infarction. After multivariable adjustment neither benefit nor rebound were apparent after study drug discontinuation, or after 3 days in all regions, except in L. America. In general, the differences in outcome and treatment effect were greatest when the protocol definition of myocardial infarction (CK(-MB) >1 upper normal limit) was applied. Under stricter definitions, these differences became smaller and disappeared with the investigator's assessment. CONCLUSION: The analysis suggests that the apparent differences in patient outcome and eptifibatide treatment effect can be explained largely by differences in baseline demographics and adjunctive treatment strategies as well as by the methodology of myocardial infarction definition and the adjudication process. PMID- 10666352 TI - Very early diagnosis and risk stratification of patients admitted with suspected acute myocardial infarction by the combined evaluation of a single serum value of cardiac troponin-T, myoglobin, and creatine kinase MB(mass) AB - AIMS: The diagnostic and prognostic capacity of biochemical markers of acute myocardial infarction in the emergency department were evaluated in consecutive patients (n=155) with suspected acute myocardial infarction. METHODS AND RESULTS: Serum myoglobin >/=110 microg. l(-1)and creatine kinase MB(mass)>/=5 microg. l( 1)had a high accuracy (0.77-0.85) (ns) for acute myocardial infarction diagnosis in patients presenting >2 h after symptom onset. Troponin-T (>/=0.10 microg. l( 1)) had a lower accuracy (0.53-0.70) for acute myocardial infarction diagnosis, but was the most important 1-year prognostic marker (cardiac death or non-fatal acute myocardial infarction). In patients without ST elevation, combined analysis of two biochemical tests would accurately identify an additional 20% of acute myocardial infarction patients (predictive value of a positive test=0.82) and also identify those without acute myocardial infarction (predictive value of a negative test=0.80). One-year event-free survival was excellent (96%) for patients with two negative biochemical tests, intermediate (74%) for those with discordant tests, and only 53% for patients with two positive biochemical tests. CONCLUSIONS: Analysis of biochemical tests in the emergency department prior to hospital admission could accurately identify approximately 20% additional acute myocardial infarction patients. The prognosis of these patients is poor, and they may be a target for primary PTCA or new early initiated aggressive medical therapies. PMID- 10666353 TI - Aortic stiffness as a risk factor for recurrent acute coronary events in patients with ischaemic heart disease. AB - AIMS: Aortic elasticity is an important determinant of left ventricular performance and coronary blood flow. Moreover, it has been shown that aortic elastic properties deteriorate in patients with coronary artery disease. However, the predictive role of aortic elasticity in the occurrence of coronary events, has not been addressed so far. Therefore, we set out to test prospectively the hypothesis that invasive as well as non-invasive measures of aortic elastic properties, assessed at rest from pressure-diameter relationships, could predict the development of recurrent coronary events. METHODS AND RESULTS: Clinical variables and measures of aortic function were assessed in 54 normotensive patients with coronary artery disease. The aortic pressure-diameter relationship was derived invasively with a high-fidelity Y shaped catheter (developed in our Institution) for aortic diameter measurements, simultaneously with a Millar catheter for aortic pressure measurements. Aortic root distensibility was assessed by non-invasive techniques. During an average of 3 years follow-up, 12 of 54 patients either developed unstable angina (n=8) or acute myocardial infarction (n=4). By multivariate Cox model analysis, aortic stiffness was the strongest predictor of progression to any end-point (relative risk: 3.24, CI: 1.79 to 5.83;P=0.000). When aortic stiffness was not considered, aortic distensibility was the only independent predictor for acute coronary syndromes (relative risk: 0.37 CI: 0.21 to 0.65;P=0.000). CONCLUSION: In patients with coronary artery disease, aortic elastic properties are powerful and independent risk factors for recurrent acute coronary events. PMID- 10666354 TI - Safety and prognostic value of early dobutamine-atropine stress echocardiography in patients with spontaneous chest pain and a non-diagnostic electrocardiogram. AB - AIMS: To risk stratify and shorten hospital stay in patients with spontaneous (resting) chest pain and a non-diagnostic electrocardiogram (ECG). METHODS AND RESULTS: The study comprised 102 patients (mean age 58+/-12 years, 67 men) with spontaneous chest pain and a non-diagnostic ECG. Forty-three patients had suspected coronary artery disease and 59 had known (but of unknown actual significance) coronary artery disease. All patients underwent serial creatine kinase enzyme measurements, continuous ECG monitoring for at least 12 h and early dobutamine-atropine stress echocardiography in patients with negative creatine kinase enzymes and normal findings at ECG monitoring. Dobutamine-atropine stress echocardiography was considered positive in patients with new or worsening wall thickening abnormalities. Patients with negative dobutamine-atropine stress echocardiography were discharged after the test. In-hospital and 6 month follow up events noted were cardiac death, non-fatal myocardial infarction, unstable angina, and coronary artery bypass surgery or angioplasty. Thirteen patients had evidence of evolving myocardial infarction by elevated creatine kinase enzymes, or unstable angina by ECG monitoring. In the remaining 89 patients, dobutamine atropine stress echocardiography was performed after a median observation period of 31 h (range 12-68 h). During dobutamine-atropine stress echocardiography no serious complications (death, non-fatal myocardial infarction, sustained ventricular tachycardia or ventricular fibrillation) occurred. Dobutamine atropine stress echocardiography results were of poor quality in three, non diagnostic in six, negative in 44 and positive in 36 patients. In the 80 patients with diagnostic dobutamine-atropine stress echocardiography, variables associated with in-hospital events (n=7) were history of exertional angina (P<0. 005), chest pain score (P<0.005), stress-induced angina (P<0.001) and positive dobutamine atropine stress echocardiography (P<0.005). Variables associated with follow-up events (n=11) were history of exertional angina (P<0.05), chest pain score (P<0.001), stress-induced angina (P<0.01) and positive dobutamine-atropine stress echocardiography (P<0.01). At multivariate analysis the only significant predictor of events was positive dobutamine-atropine stress echocardiography (P<0.01). CONCLUSION: Early dobutamine-atropine stress echocardiography may safely distinguish between low- and high-risk subsets for subsequent cardiac events in patients with spontaneous chest pain and a non-diagnostic ECG. PMID- 10666355 TI - Cerebrovascular reactivity is impaired in patients with cardiac failure. AB - AIMS: We undertook this study to evaluate potential changes in cerebral vasoreactivity in patients with cardiac failure and their consequent dependence upon cardiac functional variables. METHODS AND RESULTS: A total of 50 patients with various degrees of heart failure, 20 age-matched controls and 20 normal controls were examined. Cerebrovascular reactivity was examined with the carbon dioxide technique. Mean flow velocities of both middle cerebral arteries as well as end-tidal carbon dioxide partial pressure were continuously registered. Normal controls were examined on two different occasions, to evaluate the technique's reproducibility. Cerebrovascular reactivity was significantly reduced in all examined patients as compared to controls, and in NYHA IV as compared to NYHA II and III patients. A significant relationship between cerebrovascular reactivity and left ventricular ejection fraction was evident. Reproducibility of the technique was satisfactory. CONCLUSION: Our study provided evidence of significantly reduced cerebrovascular reactivity in patients with cardiac failure, which was significantly related to the NYHA grade and the left ventricular ejection fraction. PMID- 10666357 TI - ESC news and appointments PMID- 10666356 TI - Blood pressure awareness in Austria. A 20-year evaluation, 1978-1998. AB - AIM: To evaluate public awareness following a nationwide educational campaign on hypertension. METHODS AND RESULTS: In 1978 the Austrian Heart Foundation conducted a nationwide educational campaign to increase the awareness of the population regarding the importance of recognizing and treating high blood pressure. Following this campaign, five opinion polls of random and representative samples were conducted to measure the awareness and knowledge of the population relating to issues of high blood pressure. The poll results indicated that during the period immediately following the awareness campaign, knowledge and perception of the dangers of high blood pressure increased. However, this effect dissipated during subsequent years. In 1978, 14% of the population were reported to be hypertensive and 10% had no relevant information about their blood pressure status. In 1998, those who labelled themselves as hypertensive dropped to 12%, while those who did not know their blood pressure values, increased to 14%. The percentage of the population who recalled having their blood pressure measured during the last 3 months dropped from 49% in 1978, to 34% in 1993, and remained at 34% in 1998. CONCLUSION: We conclude that the intensive blood pressure education campaign had only a temporary effect on improving blood pressure awareness. Improved strategies are needed to achieve better community control of hypertension. PMID- 10666358 TI - Autoregressive models for estimating phylogenetic and environmental effects: accounting for within-species variations. AB - Comparative methods are used to investigate the attributes of present species or higher taxa. Difficulties arise from the phylogenetic heritage: taxa are not independent and neglecting phylogenetic inertia can lead to inaccurate results. Within-species variations in life-history traits are also not negligible, but most comparative methods are not designed to take them into account. Taxa are generally described by a single value for each trait. We have developed a new model which permits the incorporation of both the phylogenetic relationships among populations and within-species variations. This is an extension of classical autoregressive models. This family of models was used to study the effect of fishing on six demographic traits measured on 77 populations of teleost fishes. PMID- 10666359 TI - Stochastic variation in food availability influences weight and age at maturity. AB - Variation in mean food availability, and in the variance around the mean, affects the growth rate during development. Previous theoretical work on the influence of environmental quality or growth rates on the phenotypic traits age and size at maturation assumed that there is no variation in growth rate or food availability within a generation. We develop a stochastic dynamic programming (SDP) model of the foraging behaviour of aphidophagous syrphids, and use this model to predict when syrphids should pupate (mature) when average food availability changes, or varies stochastically, during development. The optimal strategy takes into account not only the availability of food, but also the timing of its availability. Food availability, when small, influences developmental time, but not weight at pupation. Food availability, when large, influences weight at pupation, but not developmental time. When the food supply is low, the optimal strategy adjusts the size at pupation downwards for stochastic as opposed to deterministic availability of food. The conclusions reinforce the need for life history studies to consider state dependence and short-term variability in growth rates. PMID- 10666360 TI - Distributions of dimeric tandem repeats in non-coding and coding DNA sequences. AB - We study the length distribution functions for the 16 possible distinct dimeric tandem repeats in DNA sequences of diverse taxonomic partitions of GenBank (known human and mouse genomes, and complete genomes of Caenorhabditis elegans and yeast). For coding DNA, we find that all 16 distribution functions are exponential. For non-coding DNA, the distribution functions for most of the dimeric repeats have surprisingly long tails, that fit a power-law function. We hypothesize that: (i) the exponential distributions of dimeric repeats in protein coding sequences indicate strong evolutionary pressure against tandem repeat expansion in coding DNA sequences; and (ii) long tails in the distributions of dimers in non-coding DNA may be a result of various mutational mechanisms. These long, non-exponential tails in the distribution of dimeric repeats in non-coding DNA are hypothesized to be due to the higher tolerance of non-coding DNA to mutations. By comparing genomes of various phylogenetic types of organisms, we find that the shapes of the distributions are not universal, but rather depend on the specific class of species and the type of a dimer. PMID- 10666361 TI - Specific and non-specific defense against parasitic attack. AB - Specific defense protects against some parasite genotypes but not others, whereas non-specific defense is effective against all genotypes of a parasite. Some empirical studies observe hosts with variability only in non-specific defense, other studies find only specific defense. I analyse a model with combined specific and non-specific defense to determine the conditions that favor detectable variation in each form of defense. High variation in non-specific defense is often maintained when resistance increases in an accelerating way with investment, whereas low variation tends to occur when resistance increases at a decelerating rate with investment. Variation in specific defense rises as the parasite pays a higher cost to attack a broad host range (high cost of virulence), as the number of alternative specificities declines, and as the average level of non-specific defense increases. The last condition occurs because greater non-specific protection tends to stabilize the gene frequency dynamics of specific defense. Selection favors a negative association between costly components of specific and non-specific defense-hosts defended by one component are favored if they have reduced allocation to other costly components. A negative association confounds the measurement of costs of resistance. Individuals with specific defense may have reduced investment in costly non specific defense. This leads to an apparent advantage of specifically defended hosts in the absence of parasites and a measured cost of resistance that is negative. PMID- 10666362 TI - Is there replication-associated mutational pressure in the Saccharomyces cerevisiae genome? AB - Compositional bias of yeast chromosomes was analysed using detrended DNA walks. Unlike eubacterial chromosomes, the yeast chromosomes did not show the specific asymmetry correlated with origin and terminus of replication. It is probably a result of a relative excess of autonomously replicating sequences (ARS) and of random choice of these sequences in each replication cycle. Nevertheless, the last ARS from both ends of chromosomes are responsible for unidirectional replication of subtelomeric sequences with pre-established leading/lagging roles of DNA strands. In these sequences a specific asymmetry is observed, resembling the asymmetry introduced by replication-associated mutational pressure into eubacterial chromosomes. PMID- 10666363 TI - Vasculogenic mimicry in tumors. Fact or artifact? PMID- 10666365 TI - Vasculogenic mimicry: how convincing, how novel, and how significant? PMID- 10666364 TI - Vasculogenic mimicry and tumor angiogenesis. AB - Tumors require a blood supply for growth and hematogenous dissemination. Much attention has been focused on the role of angiogenesis-the recruitment of new vessels into a tumor from pre-existing vessels. However, angiogenesis may not be the only mechanism by which tumors acquire a microcirculation. Highly aggressive and metastatic melanoma cells are capable of forming highly patterned vascular channels in vitro that are composed of a basement membrane that stains positive with the periodic acid-Schiff (PAS) reagent in the absence of endothelial cells and fibroblasts. These channels formed in vitro are identical morphologically to PAS-positive channels in histological preparations from highly aggressive primary uveal melanomas, in the vertical growth phase of cutaneous melanomas, and in metastatic uveal and cutaneous melanoma. The generation of microvascular channels by genetically deregulated, aggressive tumor cells was termed "vasculogenic mimicry" to emphasize their de novo generation without participation by endothelial cells and independent of angiogenesis. Techniques designed to identify the tumor microcirculation by the staining of endothelial cells may not be applicable to tumors that express vasculogenic mimicry. Although it is not known if therapeutic strategies targeting endothelial cells will be effective in tumors whose blood supply is formed by tumor cells in the absence of angiogenesis, the biomechanical and molecular events that regulate vasculogenic mimicry provide opportunities for the development of novel forms of tumor targeted treatments. The unique patterning characteristic of vasculogenic mimicry provides an opportunity to design noninvasive imaging techniques to detect highly aggressive neoplasms and their metastases. PMID- 10666366 TI - Differential growth: from carcinogenesis to liver repopulation. PMID- 10666367 TI - Control of apoptosis during angiogenesis by survivin expression in endothelial cells. AB - Mechanisms controlling endothelial cell survival during angiogenesis were investigated. Stimulation of quiescent endothelial cells with mitogens, including vascular endothelial growth factor and basic fibroblast growth factor, induced up to approximately 16-fold up-regulation of the cell cycle-regulated apoptosis inhibitor survivin. Mitogen stimulation rapidly increased survivin RNA expression in endothelial cells, which peaked after 6 to 10 hours in culture and decreased by 24 hours. Inflammatory cytokines, tumor necrosis factor alpha, and interleukin 1 did not induce survivin expression in endothelial cells. Formation of three dimensional vascular tubes in vitro was associated with strong induction of survivin in endothelial cells, as compared with two-dimensional cultures. By immunohistochemistry, survivin was minimally expressed in endothelium of nonproliferating capillaries of normal skin, whereas it became massively up regulated in newly formed blood vessels of granulation tissue in vivo. Recombinant expression of green fluorescent protein survivin in endothelial cells reduced caspase-3 activity and counteracted apoptosis induced by tumor necrosis factor alpha/cycloheximide. These findings identify survivin as a novel growth factor-inducible protective gene expressed by endothelial cells during angiogenesis. Therapeutic manipulation of survivin expression and function in endothelium may influence compensatory or pathological (tumor) angiogenesis. PMID- 10666368 TI - Strong immunostaining for myogenin in rhabdomyosarcoma is significantly associated with tumors of the alveolar subclass. AB - Rhabdomyosarcomas are a heterogeneous group of tumors with respect to their molecular basis, degree of differentiation, histology, and clinical behavior. Because of the wide variation of tumor morphology, it is often difficult to distinguish between the distinct subtypes of rhabdomyosarcomas. By using cryosections of tumor specimens and immunohistochemistry, in the present study we show that strong expression of myogenin in rhabdomyosarcoma is associated with alveolar histology (P = <0.0001, Fisher's exact test). Although staining for myogenin was observed in 22 of 26 rhabdomyosarcomas, all alveolar rhabdomyosarcomas (nine of nine) showed high levels of staining for myogenin, as defined by the frequency and intensity of staining of the tumor cells. The staining pattern suggests that the tumor cells are clonally derived from myogenin positive progenitor cells. In contrast, most embryonal rhabdomyosarcomas (13 of 15) were either negative or showed a low level of staining for myogenin. In these tumors a larger proportion of tumor cells were distinctly negative for myogenin. Six of seven alveolar rhabdomyosarcomas that strongly stained for myogenin were also positive for Pax3-7/Forkhead (FKHR) by polymerase chain reaction/reverse transcriptase-polymerase chain reaction. One of two embryonal rhabdomyosarcomas that strongly stained for myogenin was retrospectively found to be positive for Pax3/FKHR transcripts. Quantitative analysis for myogenin by Western blotting using a smaller subset of rhabdomyosarcomas revealed that in general there was a good correlation between immunohistochemical staining and Western blotting (P = 0.01, Pearson Correlation), although the former technique was more sensitive for detecting tumors with low levels of the protein. On average, alveolar rhabdomyosarcomas expressed at least threefold more myogenin than embryonal rhabdomyosarcomas. Our data show that staining for myogenin will be a simple, rapid, and accurate adjunct for distinguishing between alveolar and embryonal rhabdomyosarcomas. We propose that embryonal rhabdomyosarcomas result from an early block in myogenesis, before the expression of myogenin. In contrast, we propose that alveolar rhabdomyosarcomas either originate from a late block in myogenesis (after expression of myogenin) or that the pathological mechanisms involved in these neoplasms also induce strong expression of this protein. PMID- 10666369 TI - Bcl-2 and p53 protein expression, apoptosis, and p53 mutation in human epithelial ovarian cancers. AB - Bcl-2 and p53 gene products have been both linked to cell death by apoptosis. In the present study, we examined the relationship of Bcl-2 and p53 protein expression, p53 mutation and apoptosis in normal human ovaries and different types of human ovarian epithelial tumors by immunohistochemical localization, in situ terminal transferase-mediated dUTP nick end labeling and polymerase chain reaction-single strand conformation polymorphism. It was found that Bcl-2 expressed strongly in the surface epithelium of normal ovaries and benign and borderline ovarian tumors but weakly in the malignant tumors. On the contrary, strong protein expression of p53 was found in 54% (25/46) of the malignant epithelial tumors examined but similar expression of p53 was not observed in borderline and benign tumors and normal ovarian surface epithelium. A significant inverse correlation between Bcl-2 and p53 expression was found in the malignant ovarian tumors examined. p53 gene mutation at exons 5-11 was however not a pre requisite for p53 expression in both borderline and malignant tumors. Apoptotic activities, as reflected by apoptotic indices, were low in normal ovarian surface epithelium and benign tumors but were increased in borderline and malignant tumors, with the highest average apoptotic index found in grade III malignant tumors. Statistical analyses showed a positive correlation between apoptosis and p53 expression, but similar correlation was not found between apoptosis and Bcl-2 expression. Our results also indicate that although expression of Bcl-2 is important during ovarian carcinogenesis, the Bcl-2 protein may have other roles to play apart from being a modulator of apoptosis in human ovarian epithelial cancers. PMID- 10666370 TI - Loss of FHIT expression in transitional cell carcinoma of the urinary bladder. AB - Cytogenetic and loss of heterozygosity (LOH) studies demonstrated chromosome 3p deletions in transitional cell carcinoma (TCC). We recently cloned the tumor suppressor gene FHIT (fragile histidine triad) at 3p14.2, one of the most frequently deleted chromosomal regions in TCC of the bladder, and showed that it is the target of environmental carcinogens. Abnormalities at the FHIT locus have been found in tumors of the lung, breast, cervix, head and neck, stomach, pancreas, and clear cell carcinoma of the kidney. We examined six TCC derived cell lines (SW780, T24, Hs228T, CRL7930, CRL7833, and HTB9) and 30 primary TCC of the bladder for the integrity of the FHIT transcript, using reverse transcriptase polymerase chain reaction (RT-PCR) to investigate a potential role of the FHIT gene in TCC of the bladder. In addition, we tested expression of the Fhit protein in the six TCC-derived cell lines by Western blot analysis and in 85 specimens of primary TCCs by immunohistochemistry. Three of the six cell lines (50%) did not show the wild-type FHIT transcript, and Fhit protein was not detected in four of the six cell lines (67%) tested. Fhit expression also was correlated with pathological and clinical status. A significant correlation was observed between reduced Fhit expression and advanced stage of the tumors. Overall, 26 of 30 (87%) primary TCCs showed abnormal transcripts. Fhit protein was absent or greatly reduced in 61% of the TCCs analyzed by immunohistochemistry. These results suggested that loss of Fhit expression may be as important in the development of bladder cancer as it is for other neoplasms caused by environmental carcinogens. PMID- 10666371 TI - Genetic profile of gliosarcomas. AB - There are distinct genetic pathways leading to the glioblastoma, the most malignant astrocytic brain tumor. Primary (de novo) glioblastomas develop in older patients and are characterized by epidermal growth factor (EGF) receptor amplification/overexpression, p16 deletion, and PTEN mutations, whereas secondary glioblastomas that progressed from low-grade or anaplastic astrocytoma develop in younger patients and frequently contain p53 mutations. In this study, we assessed the genetic profile of gliosarcoma, a rare glioblastoma variant characterized by a biphasic tissue pattern with alternating areas displaying glial and mesenchymal differentiation. Single-strand conformation polymorphism followed by direct DNA sequencing revealed p53 mutations in five of 19 gliosarcomas (26%) and PTEN mutations in seven cases (37%). Homozygous p16 deletion was detected by differential polymerase chain reaction in seven (37%) gliosarcomas. The overall incidence of alterations in the Rb pathway (p16 deletion, CDK4 amplification, or loss of pRb immunoreactivity) was 53%, and these changes were mutually exclusive. Coamplification of CDK4 and MDM2 was detected in one gliosarcoma. None of the gliosarcomas showed amplification or overexpression of the EGF receptor. Thus gliosarcomas exhibit a genetic profile similar to that of primary (de novo) glioblastomas, except for the absence of EGFR amplification/overexpression. Identical PTEN mutations in the gliomatous and sarcomatous tumor components were found in two cases. Other biopsies contained p16 deletions, an identical p53 mutation, or coamplification of MDM2 and CDK4 in both tumor areas. This strongly supports the concept of a monoclonal origin of gliosarcomas and an evolution of the sarcomatous component due to aberrant mesenchymal differentiation in a highly malignant astrocytic neoplasm. PMID- 10666372 TI - APC mutations in sporadic medulloblastomas. AB - The cerebellar medulloblastoma (WHO Grade IV) is a highly malignant, invasive embryonal tumor with preferential manifestation in children. Several molecular alterations appear to be involved, including isochromosome 17q and the p53, PTCH, and beta-catenin gene mutations. In this study, 46 sporadic medulloblastomas were screened for the presence of mutations in genes of the Wnt signaling pathway (APC and beta-catenin). Single-strand conformational polymorphism (SSCP) analysis followed by direct DNA sequencing revealed 3 miscoding APC mutations in 2 (4.3%) medulloblastomas. One case contained a GCA-->GTA mutation at codon 1296 (Ala- >Val), and another case had double point mutations at codons 1472 (GTA-->ATA, Val ->Ile) and 1495 (AGT-->GGT, Ser-->Gly). Miscoding beta-catenin mutations were detected in 4 tumors (8.7%). Three of these were located at codon 33 (TCT -->TTT, Ser-->Phe) and another at codon 37 (TCT-->GCT, Ser-->Ala). Adenomatous polyposis coli (APC) gene and beta-catenin mutations were mutually exclusive and occurred in a total of 6 of 46 cases (13%). Although germline APC mutations are a well established cause of familial colon and brain tumors (Turcot syndrome), this study provides the first evidence that APC mutations are also operative in a subset of sporadic medulloblastomas. PMID- 10666373 TI - In vitro induction of giant cell tumors from cultured hamster islets treated with N-Nitrosobis(2-Oxopropyl)amine. AB - Giant cell carcinoma of the pancreas is a rare tumor. Its histogenesis is still controversial. In a Syrian hamster pancreatic cancer model, tumors similar to human giant cell carcinomas have been induced at an extremely low rate of incidence and after the use of high doses of pancreatic carcinogens. Thus far no tumors of giant cell type have been induced by the in vitro treatment of hamster pancreatic ductal cells with the potent pancreatic carcinogen N-nitrosobis(2 oxopropyl)amine (BOP). In the present study we report the induction of giant cell carcinoma from hamster islets treated with BOP in vitro. The results suggest that in hamsters some component of islet cells, probably stem cells, are the origin of giant cell carcinoma. PMID- 10666374 TI - Sensitive immunoassay of tissue cell proteins procured by laser capture microdissection. AB - Coupling laser capture microdissection (LCM) with sensitive quantitative chemiluminescent immunoassays has broad applicability in the field of proteomics applied to normal, diseased, or genetically modified tissue. Quantitation of the number of prostate-specific antigen (PSA) molecules/cell was conducted on human prostate tissue cells procured by LCM from fixed and stained frozen sections. Under direct microscopic visualization, laser shots 30 microm in diameter captured specific cells from the heterogeneous tissue section onto a polymer transfer surface. The cellular macromolecules from the captured cells were solubilized in a microvolume of extraction buffer and directly assayed using an automated (1.5 hour) sandwich chemiluminescent immunoassay. Calibration of the chemiluminescent assay was conducted by developing a standard curve using known concentrations of PSA. After the sensitivity, precision, and linearity of the chemiluminescent assay was verified for known numbers of solubilized microdissected tissue cells, it was then possible to calculate the number of PSA molecules per microdissected tissue cell for case samples. In a study set of 20 cases, using 10 replicate samples of 100 laser shots per sample, the within-run (intraassay) SD was approximately 10% of the mean or less for all cases. In this series the number of PSA molecules per microdissected tissue cell ranged from 2 x 10(4) to 6. 3 x 10(6) in normal epithelium, prostate intraepithelial neoplasia (PIN), and invasive carcinoma. Immunohistochemical staining of human prostate for PSA was compared with the results of the soluble immunoassay for the same prostate tissue section. Independent qualitative scoring of anti-PSA immunohistochemical staining intensity paralleled the LCM quantitative immunoassay for each tissue subpopulation and verified the heterogeneity of PSA content between tissue subpopulations in the same case. Extraction buffers were successfully adapted for both secreted and membrane-bound proteins. This technology has broad applicability for the quantitation of protein molecules in pure populations of tissue cells. PMID- 10666375 TI - alpha5beta1 integrin expression and luminal edge fibronectin matrix assembly by smooth muscle cells after arterial injury. AB - Fibronectin is secreted from the cell as a soluble protein that must then polymerize to regulate cell function. To elucidate the process of fibronectin matrix assembly in vascular disease, we immunostained sections of balloon-injured rat carotid artery for the fibronectin-binding alpha5beta1 integrin. Whereas alpha5beta1 integrin was not evident in the normal carotid artery, its expression was induced after a vascular injury. By 14 days, the alpha5beta1 integrin was localized exclusively to the less differentiated smooth muscle cells (SMCs) at the luminal surface of the neointima. Platelet-derived growth factor-BB, dominant in neointimal formation, selectively increased the expression of the alpha5beta1 integrin by human SMCs in culture. To track the assembly of fibronectin fibers, fluorescence-labeled soluble fibronectin protomers were added to cultured SMCs and to fresh segments of normal and balloon-injured rat carotid arteries. Fibronectin fiber formation in cultured SMCs could be detected within 10 minutes, and was blocked by an RGD peptide, an anti-beta1 integrin antibody, and an anti alpha5beta1 integrin antibody, but not by an anti-beta3 integrin antibody. En face confocal microscopy of arterial segments revealed that soluble fibronectin had polymerized on the alpha5beta1 integrin-expressing SMCs of the luminal surface of the injured arterial neointima, but not on the alpha5beta1 integrin negative neointimal SMCs below this or on the endothelial cells of uninjured arteries. Furthermore, in situ fibronectin assembly by the neointimal SMCs was inhibited by an RGD peptide and by an anti-beta1 integrin antibody. These studies indicate that a subpopulation of SMCs in the repairing artery wall orchestrates integrin-mediated fibronectin assembly. PMID- 10666376 TI - Type VIII collagen stimulates smooth muscle cell migration and matrix metalloproteinase synthesis after arterial injury. AB - Type VIII collagen is a matrix protein expressed in a number of tissues undergoing active remodeling, including injured arteries during neointimal formation and in human atherosclerotic plaques; however, very little is known about its function. We have investigated whether the type VIII collagen stimulates smooth muscle cell (SMC) migration and invasion by binding to integrin receptors and up-regulating matrix metalloproteinase (MMP) production. SMCs attached to plates coated with type VIII collagen in a dose-dependent manner, with maximal attachment occurring with coating solutions containing 25 microgram/ml collagen. Type VIII collagen at 100 microgram/ml stimulated an 83 fold increase in the migration of SMCs in a chemotaxis chamber. Antibodies against beta1 integrin receptors prevented attachment and migration of SMCs. Antibodies against alpha1 or alpha2 integrins reduced attachment of SMCs to type VIII collagen by 29% and 77%, respectively. We found that SMCs grown from the rat neointima, but not medial SMCs, increased their production of MMP-2 and -9 on adherence to type VIII collagen. This suggests that there is an important difference in phenotype between intimal and medial SMCs and that intimal SMCs have distinct matrix-dependent signaling mechanisms. Our findings suggest that type VIII collagen deposited in vascular lesions functions to promote SMC attachment and chemotaxis, and signals through integrin receptors to stimulate MMP synthesis, all of which are important mechanisms used in cell migration and invasion. PMID- 10666377 TI - Platelet-derived growth factor-A-induced retinal gliosis protects against ischemic retinopathy. AB - Retinal astrocytes are located in the nerve fiber layer and along retinal blood vessels and have been hypothesized to participate in the induction and maintenance of the blood-retinal barrier. Platelet-derived growth factor-A (PDGF A) is normally produced by retinal ganglion cells and is involved in astrocyte recruitment and proliferation. We used gain-of-function transgenic mice that express PDGF-A in photoreceptors to explore the roles of PDGF-A and astrocytes in the retina. Transgene-positive mice developed glial infiltration of the inner retina and had significantly less oxygen-induced retinal vascular closure and no neovascularization compared with littermate controls, which had prominent vascular closure and neovascularization. The increased survival of endothelial cells in transgenic mice in the face of oxygen-induced down-regulation of vascular endothelial growth factor was accompanied by an increase in astrocyte derived fibroblast growth factor-2. Therefore, PDGF-A increases retinal astrocytes, which promote the survival of endothelial cells as well as their expression of barrier characteristics. PMID- 10666378 TI - Temporal accrual of complement proteins in amyloid plaques in Down's syndrome with Alzheimer's disease. AB - The complement system constitutes a series of enzymatic steps involved in the inflammatory response and is activated in Alzheimer's disease (AD). Using Down's syndrome (DS) brains as a temporal model for the progression of AD, we examined components of the complement cascade and their relationship to other principal events in AD pathology: Abeta42 deposition, neuritic changes, neurofibrillary tangles (NFTs), and gliosis (reactive astrocytes, activated microglia). Adjacent sections of frontal cortex from 24 DS subjects ranging in age from 12 to 73 years were immunohistochemically examined for immunoreactivity (IR) of classical complement proteins (Clq and C3), markers indicating activation of complement (C4d and C5b-9), the complement inhibitor apolipoprotein J (apo J), and markers of AD neuropathology. Abeta42-labeled diffuse plaques were first detected in a 12 year-old DS subject and were not labeled by any of the complement antibodies. Colocalization of Abeta42 with Clq, C3, C4d, and/or apo J was first detected in compacted plaques in the brain of a 15-year-old DS patient with features of mature AD pathology, such as reactive astrocytes, activated microglia, dystrophic neurites, and a few NFTs. IR for C4d and C5b-9 (membrane attack complex, MAC) was observed in small numbers of plaque-associated dystrophic neurites and in focal regions of pyramidal neurons in this 15-year-old. The only other young ((max) and on the shape of the foot of the transmembrane action potential (V(m) foot). Resistive discontinuities primarily affect f1.gif" BORDER="0">(max), and an additional capacitive component in the local circuit due to the capillaries in interstitial space primarily affects V(m) foot. Resistive discontinuities also have an important influence on cardiac conduction. These discontinuities include spatial variations in the size of interstitial space (interstitial resistive discontinuities) and the role of cellular scaling (effects of cell size) when changes occur in the cellular and multicellular distribution of gap junctions during remodeling of normal mature myocardium to proarrhythmic structural substrates. The full text of this article is available at http://www.circresaha.org. PMID- 10666423 TI - Mice lacking the vascular endothelial growth factor-B gene (Vegfb) have smaller hearts, dysfunctional coronary vasculature, and impaired recovery from cardiac ischemia. AB - Vascular endothelial growth factor-B (VEGF-B) is closely related to VEGF-A, an effector of blood vessel growth during development and disease and a strong candidate for angiogenic therapies. To further study the in vivo function of VEGF B, we have generated Vegfb knockout mice (Vegfb(-/-)). Unlike Vegfa knockout mice, which die during embryogenesis, Vegfb(-/-) mice are healthy and fertile. Despite appearing overtly normal, Vegfb(-/-) hearts are reduced in size and display vascular dysfunction after coronary occlusion and impaired recovery from experimentally induced myocardial ischemia. These findings reveal a role for VEGF B in the development or function of coronary vasculature and suggest potential clinical use in therapeutic angiogenesis. PMID- 10666424 TI - Tetrahydrobiopterin improves endothelium-dependent vasodilation in chronic smokers : evidence for a dysfunctional nitric oxide synthase. AB - Conditions associated with impaired nitric oxide (NO) activity and accelerated atherosclerosis have been shown to be associated with a reduced bioavailability of tetrahydrobiopterin (BH4). We therefore hypothesized that BH4 supplementation may improve endothelial dysfunction of chronic smokers. Forearm blood flow (FBF) responses to the endothelium-dependent vasodilators acetylcholine (ACh; 0.75, 1.5, and 3.0 microg/100 mL tissue/min) or serotonin (5-HT; 0.7, 2.1, and 6.3 ng/100 mL tissue/min), to the inhibitor of endothelial nitric oxide synthase (NOS) N(G)-monomethyl-L-arginine (L-NMMA; 2, 4, and 8 micromol/min), and to the endothelium-independent vasodilator sodium nitroprusside (SNP; 0.1, 0.3, and 1.0 microg/100 mL tissue/min) were measured by venous occlusion plethysmography in controls and chronic smokers. Drugs were infused into the brachial artery, and FBF was measured before and during concomitant intra-arterial infusion of BH4, tetrahydroneopterin (NH4; another reduced pteridine), or the antioxidant vitamin C (6 and 18 mg/min). In control subjects, BH4 had no effect on FBF in response to ACh, 5-HT, and SNP. In contrast, in chronic smokers, the attenuated FBF responses to ACh and 5-HT were markedly improved by concomitant administration of BH4, whereas the vasodilator responses to SNP were not affected. L-NMMA-induced vasoconstriction was significantly reduced in smokers compared with controls, suggesting impaired basal NO bioactivity. BH4 improved L-NMMA responses in smokers while having no effect on L-NMMA responses in controls. Pretreatment with vitamin C abolished BH4 effects on ACh-dependent vasodilation. In vitro, NH4 scavenged superoxide created by the xanthine/xanthine oxidase reaction equipotent like BH4 but failed to modify ACh-induced changes in FBF in chronic smokers in vivo. These data support the concept that in addition to the free radical burden of cigarette smoke, a dysfunctional NOS III due to BH4 depletion may contribute at least in part to endothelial dysfunction in chronic smokers. PMID- 10666425 TI - Formation of heterotypic gap junction channels by connexins 40 and 43. AB - Gap junctions formed between transfected cells expressing connexin (Cx) 40 and Cx43 (Cx43-RIN, Cx40-HeLa, and Cx43-HeLa) revealed a relationship, g(j)=f(V(j)), at steady state, that is typified by a nonsymmetrical behavior similar to that previously reported for other heterotypic channels (gap junction conductance [g(j)]; transjunctional voltage [V(j)]). The unitary conductance of the channels was sensitive to the polarity of V(j). A main state conductance of 61 pS was found when the Cx43 cell was stepped positively or the Cx40 cell negatively (V(j)=70 mV); the reverse polarities yielded a conductance of 100 pS. These heterotypic channels were permeable to carboxyfluorescein. In addition, two other heterotypic forms are illustrated to demonstrate that endogenous Cx45 expression cannot explain the results. The demonstration of heterotypic Cx40-Cx43 channels may have implications for the propagation of the electrical impulse in heart. For example, they may contribute to the slowing of the impulse propagation through the junctions between Purkinje fibers and ventricular muscle. PMID- 10666426 TI - Efficacy of implantable cardioverter-defibrillators for the prevention of sudden death in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: Hypertrophic cardiomyopathy is a genetic disease associated with a risk of ventricular tachyarrhythmias and sudden death, especially in young patients. METHODS: We conducted a retrospective multicenter study of the efficacy of implantable cardioverter-defibrillators in preventing sudden death in 128 patients with hypertrophic cardiomyopathy who were judged to be at high risk for sudden death. RESULTS: At the time of the implantation of the defibrillator, the patients were 8 to 82 years old (mean [+/-SD], 40+/-16), and 69 patients (54 percent) were less than 41 years old. The average follow-up period was 3.1 years. Defibrillators were activated appropriately in 29 patients (23 percent), by providing defibrillation shocks or antitachycardia pacing, with the restoration of sinus rhythm; the average age at the time of the intervention was 41 years. The rate of appropriate defibrillator discharge was 7 percent per year. A total of 32 patients (25 percent) had episodes of inappropriate discharges. In the group of 43 patients who received defibrillators for secondary prevention (after cardiac arrest or sustained ventricular tachycardia), the devices were activated appropriately in 19 patients (11 percent per year). Of 85 patients who had prophylactic implants because of risk factors (i.e., for primary prevention), 10 had appropriate interventions (5 percent per year). The interval between implantation and the first appropriate discharge was highly variable but was substantially prolonged (four to nine years) in six patients. In all 21 patients with stored electrographic data and appropriate interventions, the interventions were triggered by ventricular tachycardia or fibrillation. CONCLUSIONS: Ventricular tachycardia or fibrillation appears to be the principal mechanism of sudden death in patients with hypertrophic cardiomyopathy. In high-risk patients with hypertrophic cardiomyopathy, implantable defibrillators are highly effective in terminating such arrhythmias, indicating that these devices have a role in the primary and secondary prevention of sudden death. PMID- 10666427 TI - Prothrombin and factor V mutations in women with a history of thrombosis during pregnancy and the puerperium. AB - BACKGROUND: Venous thromboembolism is a leading cause of morbidity and mortality during pregnancy and the puerperium. However, the role of mutations in the prothrombin and factor V genes and other thrombophilic abnormalities as risk factors for thromboembolism in women during pregnancy and the pueperium is not known. METHODS: In a study of 119 women with a history of venous thromboembolism during pregnancy and the puerperium and 233 age-matched normal women, we measured the activity of antithrombin, protein C, protein S, and lupus anticoagulant. We also performed genetic analyses to detect the G1691A mutation in the factor V gene (factor V Leiden), the G20210A mutation in the prothrombin gene, and the C677T mutation in the methylenetetrahydrofolate reductase gene. Blood samples were obtained at least three months post partum or after the cessation of lactation. RESULTS: Among the women with a history of venous thromboembolism, the prevalence of factor V Leiden was 43.7 percent, as compared with 7.7 percent among the normal women (relative risk of venous thromboembolism, 9.3; 95 percent confidence interval, 5.1 to 16.9); that of the G20210A prothrombin-gene mutation, 16.9 percent as compared with 1.3 percent (relative risk, 15.2; 95 percent confidence interval, 4.2 to 52.6); and that of both factor V Leiden and the G20210A prothrombin-gene mutation 9.3 percent as compared with 0 (estimated odds ratio, 107). Assuming an overall risk of 1 in 1500 pregnancies, the risk of thrombosis among carriers of factor V Leiden was 0.2 percent, among carriers of the G20210A prothrombin-gene mutation, 0.5 percent, and among carriers of both defects, 4.6 percent, as calculated in a multivariate analysis. CONCLUSIONS: The G20210A prothrombin-gene mutation and factor V Leiden individually are associated with an increased risk of venous thromboembolism during pregnancy and the puerperium, and the risk among women with both mutations is disproportionately higher than that among women with only one mutation. PMID- 10666428 TI - Retinopathy and nephropathy in patients with type 1 diabetes four years after a trial of intensive therapy. AB - BACKGROUND: Among patients with type 1 diabetes mellitus, intensive therapy (with the aim of achieving near-normal blood glucose and glycosylated hemoglobin concentrations [hemoglobin A1c]) markedly reduces the risk of microvascular complications as compared with conventional therapy. To assess whether these benefits persist, we compared the effects of former and intensive conventional therapy on the recurrence and severity of retinopathy and nephropathy for four years after the end of the Diabetes Control and Complications Trial (DCCT). METHODS: At the end of the DCCT, the patients in the conventional-therapy group were offered intensive therapy, and the care of all patients was transferred to their own physicians. Retinopathy was evaluated on the basis of centrally graded fundus photographs in 1208 patients during the fourth year after the DCCT ended, and nephropathy was evaluated on the basis of urine specimens obtained from 1302 patients during the third or fourth year, approximately half of whom were from each treatment group. RESULTS: The difference in the median glycosylated hemoglobin values between the conventional-therapy and intensive-therapy groups during the 6.5 years of the DCCT (average, 9.1 percent and 7.2 percent, respectively) narrowed during follow-up (median during 4 years, 8.2 percent and 7.9 percent, respectively, P<0.001). Nevertheless, the proportion of patients who had worsening retinopathy, including proliferative retinopathy, macular edema, and the need for laser therapy, was lower in the intensive-therapy group than in the conventional-therapy group (odds reduction, 72 percent to 87 percent, P<0.001). The proportion of patients with an increase in urinary albumin excretion was significantly lower in the intensive-therapy group. CONCLUSIONS: The reduction in the risk of progressive retinopathy and nephropathy resulting from intensive therapy in patients with type 1 diabetes persists for at least four years, despite increasing hyperglycemia. PMID- 10666429 TI - Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A. AB - BACKGROUND: Clostridium difficile infection can result in asymptomatic carriage, mild diarrhea, or fulminant pseudomembranous colitis. We studied whether antibody responses to C. difficile toxins affect the risks of colonization, diarrhea, and asymptomatic carriage. METHODS: We prospectively studied C. difficile infections in hospitalized patients who were receiving antibiotics. Serial stool samples were tested for C. difficile colonization by cytotoxin assay and culture. Serum antibody (IgA, IgG, and IgM) levels and fecal antibody (IgA and IgG) levels against C. difficile toxin A, toxin B, and nontoxin antigens were measured by an enzyme-linked immunosorbent assay (ELISA). RESULTS: Of 271 patients, 37 (14 percent) were colonized with C. difficile at the time of admission, 18 of whom were asymptomatic carriers. An additional 47 patients (17 percent) became infected in the hospital, 19 of whom remained asymptomatic. The baseline antibody levels were similar in the patients who later became colonized and those who did not. After colonization, those who became asymptomatic carriers had significantly greater increases in serum levels of IgG antibody against toxin A than did the patients in whom C. difficile diarrhea developed (P<0.001). The adjusted odds ratio for diarrhea was 48.0 (95 percent confidence interval, 3.4 to 678) among patients with colonization who had a serum level of IgG antibody against toxin A of 3.00 ELISA units or less, as compared with patients with colonization who had a level of more than 3.00 ELISA units. CONCLUSIONS: We find no evidence of immune protection against colonization by C. difficile. However, after colonization there is an association between a systemic anamnestic response to toxin A, as evidenced by increased serum levels of IgG antibody against toxin A, and asymptomatic carriage of C. difficile. PMID- 10666430 TI - Images in clinical medicine. Internal ventricular defibrillation. PMID- 10666431 TI - Women physicians in academic medicine: new insights from cohort studies. AB - BACKGROUND: I conducted a study to determine whether women who graduate from medical schools are more or less likely than their male counterparts to pursue full-time careers in academic medicine and to advance to the senior ranks of medical school faculties. METHODS: The rates of advancement to the ranks of assistant, associate, and full professor for all U.S. medical school graduates from 1979 through 1993 and for all members of U.S. medical school faculties from 1979 through 1997 were studied. Cohorts were defined on the basis of the year of graduation from medical school, track (tenure or nontenure), and academic department. Within each cohort, the number of women who advanced to a senior rank was compared with the number that would be expected on the basis of parity between men and women, and 95 percent confidence intervals were calculated. RESULTS: Women were significantly more likely than men to pursue an academic career. During the study period, 634 more women became faculty members than expected. The numbers were higher in the older cohorts than in the younger cohorts. The numbers of women who advanced to the ranks of associate and full professor were significantly lower than the expected numbers. This was true for both tenure and nontenure tracks, even after adjustment for the department. A total of 334 fewer women advanced to associate professor than expected, and 44 fewer women advanced to full professor than expected. CONCLUSIONS: Disparities persist in the advancement of men and women on medical school faculties. However, the numbers of women physicians at all levels of academic medicine are increasing. PMID- 10666432 TI - Occupational respiratory diseases. PMID- 10666433 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 4-2000. A 64-year-old man with Cushing's syndrome and a pancreatic mass. PMID- 10666434 TI - Sudden death in hypertrophic cardiomyopathy. PMID- 10666435 TI - The challenge of thrombophilia in maternal-fetal medicine. PMID- 10666436 TI - Women in academic medicine: new insights, same sad news. PMID- 10666437 TI - Collective bargaining is the right step. PMID- 10666438 TI - White coats should not have union labels. PMID- 10666440 TI - Correction: Looking Back on the Millennium in Medicine. PMID- 10666439 TI - Correction: Canterbury Cathedral. PMID- 10666442 TI - A novel RNA-binding protein from Triturus carnifex identified by RNA-ligand screening with the newt hammerhead ribozyme. AB - The newt hammerhead ribozyme is transcribed from Satellite 2 DNA, which consists of tandemly repeated units of 330 bp. However, different transcripts are synthesized in different tissues. In all somatic tissues and in testes, dimeric and multimeric RNA transcripts are generated which, to some extent, self-cleave into monomers at the hammerhead domain. In ovaries, primarily a distinct monomeric unit is formed by transcription, which retains an intact hammerhead self-cleavage site. The ovarian monomeric RNA associates to form a 12S complex with proteins that are poorly characterised so far. In this work we identified NORA, a protein that binds the ovarian form of the newt ribozyme. We show that the newt ribozyme binds to the Escherichia coli -expressed protein, as well as to a protein of identical size that is found exclusively in newt ovaries. Also NORA mRNA was detectable only in ovary, but in neither somatic tissues nor testes. The tissue-specific expression of NORA is analogous to the ovary-specific transcription of the newt ribozyme. Although NORA was identified by its ability to bind to the newt ribozyme in the presence of a vast excess of carrier RNA, it was able to interact with certain other RNA probes. This novel RNA-binding protein does not contain any motif characteristic for RNA-binding proteins or any other known protein domain, but it shares a striking similarity with a rat resiniferatoxin-binding protein. PMID- 10666443 TI - MethTools--a toolbox to visualize and analyze DNA methylation data. AB - The Bisulfite Genomic Sequencing technique has found wide acceptance for the generation of DNA-methylation maps with single-base resolution. The method is based on the selective deamination of cytosine to uracil (and subsequent conversion to thymine via PCR), whereas 5-methylcytosine residues remain unchanged. Methylation maps are created by the comparison of bisulfite converted sequences with the untreated genomic sequence. 'MethTools' is a collection of software tools that replaces the time-consuming manual comparison process, generates graphical outputs of methylation patterns and methyl-ation density, estimates the systematic error of the experiment and searches for conserved methylated nucleotide patterns. The programs are written in Perl 5 and C, and the source code can be downloaded. All tools run independently but the programs are interfaced. Thus, a script can perform the entire analysis procedure automatically. In addition, a web-based remote analysis service is offered. Both the source code and the remote analysis are available at http://genome.imb jena.de/methtools/ PMID- 10666444 TI - Improving dideoxynucleotide-triphosphate utilisation by the hyper-thermophilic DNA polymerase from the archaeon Pyrococcus furiosus. AB - Polymerases from the Pol-I family which are able to efficiently use ddNTPs have demonstrated a much improved performance when used to sequence DNA. A number of mutations have been made to the gene coding for the Pol-II family DNA polymerase from the archaeon Pyrococcus furiosus with the aim of improving ddNTP utilisation. 'Rational' alterations to amino acids likely to be near the dNTP binding site (based on sequence homologies and structural information) did not yield the desired level of selectivity for ddNTPs. However, alteration at four positions (Q472, A486, L490 and Y497) gave rise to variants which incorporated ddNTPs better than the wild type, allowing sequencing reactions to be carried out at lowered ddNTP:dNTP ratios. Wild-type Pfu-Pol required a ddNTP:dNTP ratio of 30:1; values of 5:1 (Q472H), 1:3 (L490W), 1:5 (A486Y) and 5:1 (Y497A) were found with the four mutants; A486Y representing a 150-fold improvement over the wild type. A486, L490 and Y497 are on analpha-helix that lines the dNTP binding groove, but the side chains of the three amino acids point away from this groove; Q472 is in a loop that connects this alpha-helix to a second long helix. None of the four amino acids can contact the dNTP directly. Therefore, the increased selectivity for ddNTPs is likely to arise from two factors: (i) small overall changes in conformation that subtly alter the nucleotide triphosphate binding site such that ddNTPs become favoured; (ii) interference with a conformational change that may be critical both for the polymerisation step and discrimination between different nucleotide triphosphates. PMID- 10666445 TI - A simple in vitro Tn7-based transposition system with low target site selectivity for genome and gene analysis. AB - A robust Tn7-based in vitro transposition system is described that displays little target site selectivity, allowing the efficient recovery of many different transposon insertions in target DNAs ranging from small plasmids to cosmids to whole genomes. Two miniTn7 derivatives are described that are useful for the analysis of genes: one a derivative for making translational and transcriptional target gene fusions and the other a derivative that can generate 15 bp (5 amino acid) insertions in target DNAs (proteins). PMID- 10666446 TI - Genetic analysis of a La homolog in Drosophila melanogaster. AB - People afflicted with certain rheumatological auto-immune diseases produce autoantibodies directed against a select group of proteins such as the La auto antigen. Biochemical studies have revealed La to be a promiscuous RNA-binding protein that appears to play a role in a variety of intracellular activities such as processing and/or transport of RNA polymerase III precursor transcripts and translational regulation from internal ribosome entry sites (IRES). We have previously identified an RNA-binding protein that is a Drosophila melanogaster homolog of La (D-La) and shown that early transcript accumulation throughout the embryo is later refined to be most prevalent in the visceral mesoderm, gut, gonads and salivary glands. Here we report the first in vivo genetic characterization of a La homolog in a multicellular eukaryote. Lethality was observed in homozygous larvae harboring a small chromosomal deletion that removed the D-La gene, which was rescued by an inducible D-La cDNA transgene. This implies that D-La confers essential functions for larval development. In addition, loss of D-La function gives rise to defects in embryonic midgut morphogenesis; one of the midgut defects correlates with loss of Ultrabithorax ( Ubx ) expression along the second midgut constriction. Finally, genetic interactions between chromosomal deficiencies that remove D-La and certain Ubx alleles were demonstrated in adults. Our results support the hypothesis that D-La provides essential functions for proper Drosophila development and imply that the conserved La family of proteins may perform critical developmental functions in higher eukaryotes. PMID- 10666447 TI - Identification of a base-specific contact between the restriction endonuclease SsoII and its recognition sequence by photocross-linking. AB - A target sequence-specific DNA binding region of the restriction endonuclease Sso II was identified by photocross-linking with an oligodeoxynucleotide duplex which was substituted with 5-iododeoxy-uridine (5-IdU) at the central position of the Sso II recognition site (CCNGG). For this purpose the Sso II-DNA complex was irradiated with a helium/cadmium laser (325 nm). The cross-linking yield obtained was approximately 50%. In the presence of excess unmodified oligodeoxynucleotide or with oligode-oxynucleotides substituted with 5-IdU elsewhere, no cross-linking was observed, indicating the specificity of the cross-linking reaction. The cross linked Sso II-oligodeoxynucleotide complex was digested with chymotrypsin, a cross-linked peptide-oligodeoxy-nucleotide complex isolated and the site of cross linking identified by Edman sequencing to be Trp61. In line with this identification is the finding that the W61A variant cannot be cross-linked with the IdU-substituted oligodeoxynucleotide, shows a decrease in affinity towards DNA and is inactive in cleavage. It is concluded that the region around Trp61 is involved in specific binding of Sso II to its DNA substrate. PMID- 10666448 TI - Transcripts of the ndhH-D operon of barley plastids: possible role of unedited site III in splicing of the ndhA intron. AB - The plastid ndhH-D operon produces several transcripts containing ndhA sequence with and without its group II intron. After sequencing an 8125 bp fragment of barley plastid DNA including the ndhH-D operon, we investigated the editing splicing status of transcripts in the range 1.0-7.8 kb. Reverse transcription and sequencing of RNA bands separated by electrophoresis were used to determine C-->U editing sites. Sites I, II and IV of ndhA and site V of ndhD were edited in all transcripts analysed and, probably, were edited before any splicing had taken place. In contrast, site III of ndhA (13 bp from the 5'-end base of the second exon) was not edited in transcripts containing the intron (including the 1.7 kb intermediary transcript consisting of the intron and the second exon) but was edited in all transcripts lacking the ndhA intron. Comparison of the secondary structures of the ndhA intron and intron-second exon intermediate suggests that G pairing prevents editing of site III in transcripts containing the intron and maintains the secondary structure required for splicing. Splicing of the ndhA intron releases the site III C from pairing and, probably, brings it close to cis acting elements for editing upstream in the first exon. PMID- 10666449 TI - An 'environment to nucleus' signaling system operates in B lymphocytes: redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation. AB - The Ref-1 (also called APE or HAP1) protein is a bifunctional enzyme impacting on a wide variety of important cellular functions. It acts as a major member of the DNA base excision repair pathway. Moreover, Ref-1 stimulates the DNA-binding activity of several transcription factors (TFs) through the reduction of highly reactive cysteine residues. Therefore, it represents a mechanism that regulates eukaryotic gene expression in a fast way. However, it has been demonstrated that external stimuli directly act on Ref-1 by increasing its expression levels, a time-consuming mechanism representing a paradox in terms of rapidity of TF regulation. In this paper we demonstrate that this is only an apparent paradox. Exposure of B lymphocytes to H(2)O(2)induced a rapid and sustained increase in Ref-1 protein levels in the nucleus as evaluated by both western blot analysis and by pulse-chase experiments. A time course, two color in situ immunocytochemistry indicated that the up-regulation of Ref-1 in the nucleus at <30 min was primarily the consequence of translocation of its cytoplasmic form. This early nuclear accumulation is effective in modulating the DNA-binding activity of the B cell-specific activator protein BSAP/Pax-5. In fact, EMSA experiments demonstrate that a transient interaction with Ref-1 up-regulates the DNA-binding activity of BSAP/Pax-5. Moreover, in a co-transfection experiment, Ref-1 increased the BSAP/Pax-5 activating effect on an oligomerized BSAP/Pax-5 binding site of the CD19 promoter by 5- to 8-fold. Thus, Ref-1 mediates its effect by up-regulating the DNA-binding activity of BSAP/Pax-5, accounting for a new and fast outside/inside pathway of signaling in B cells. PMID- 10666450 TI - Transactivation and growth suppression by the gut-enriched Kruppel-like factor (Kruppel-like factor 4) are dependent on acidic amino acid residues and protein protein interaction. AB - Gut-enriched Kruppel-like factor (GKLF or KLF4) is a pleiotropic (activating and repressive) transcription factor. This study characterizes the mechanisms of transactivation by GKLF. Using a GAL4 fusion assay, the activating domain of murine GKLF was localized to the 109 amino acid residues in the N-terminus. Site directed mutagenesis showed that two adjacent clusters of acidic residues within this region are responsible for the activating effect. Transactivation by GKLF involves intermolecular interactions as demonstrated by the ability of wild-type, but not mutated, GKLF to compete with the N-terminal activation domain. In addition, wild-type adenovirus E1A, but not a mutated E1A that failed to bind p300/CBP, inhibited transactivation by the N-terminal 109 amino acids of GKLF, suggesting that p300/CBP are GKLF's interacting partners. A physical interaction between GKLF and CBP was demonstrated by glutathione- S -transferase pull-down and by in vivo co-immuno-precipitation experiments. We also showed that the two acidic amino acid clusters are essential for this interaction, since GKLF with mutations in these residues failed to co-immunoprecipitate with CBP. Importantly, the same mutations abrogated the ability of GKLF to suppress cell growth as determined by a colony suppression assay. These studies therefore provide plausible evidence for a structural and functional correlation between the transactivating and growth-suppressing effects of GKLF. PMID- 10666451 TI - Excision repair at the level of the nucleotide in the upstream control region, the coding sequence and in the region where transcription terminates of the Saccharomyces cerevisiae MFA2 gene and the role of RAD26. AB - RAD26, the yeast homologue of human CSB, has an essential role in transcription coupled repair (TCR). We have mapped the requisite of Rad26 for nucleotide excision repair (NER) within the different regions of the yeast Saccharomyces cerevisiae MFA2 gene at nucleotide resolution. Our results show that Rad26 is dispensable for enhanced NER in both the MFA2 upstream promoter, except in the TATA box region, and for enhanced NER in both strands of the active gene at a site close to the transcription termination region. As expected, it is not needed for repair of regions downstream of where transcription terminates. However, it is required for TCR in the transcription initiation and elongation regions. Our data support the hypothesis that Rad26 is required for the interchange between holo-TFIIH and a putative repairosome containing core TFIIH and other NER proteins. Close to the end of transcription, hotspots for the repair of CPDs in both the transcribed strand and the non-transcribed strand occur. This enhanced repair is independent of Rad26. Hence, TFIIH may take a form favourable for forming a repairosome without Rad26 assistance; here the organisation of the DNA during the termination of transcription may facilitate access of a repair complex to enable enhanced repair of both strands. PMID- 10666452 TI - The tetracycline-responsive promoter contains functional interferon-inducible response elements. AB - Tetracycline (tet)-responsive expression vectors allow controlled inducible expression of proteins in mammalian cells. This system is widely used for experimental research both in vivo and in vitro. In our attempts to use this system to study the antiviral effect of IFNalpha on hepatitis B virus, we discovered an unexpected feature of the tet-responsive promoter (tet promoter) of the currently available expression vectors. IFNalphawas found to stimulate tet promoter activity after transient transfection in a dose- and cell type-dependent manner. By sequence inspection, an IFNalpha-stimulated response element (ISRE) like sequence was identified in the linker regions located between the heptameric tet operator sequences. Gel shift assays revealed binding of IFN-stimulated gene factors to these sequences, indicating that they mediate the IFNalpha-mediated promoter stimulation. These data demonstrate an unexpected feature of the tet responsive expression system which needs to be taken into account when using this system for analysis of cytokine functions in vitro and in vivo. The data also imply that the tet promoter-based expression system can be rendered non responsive to IFNalpha by mutagenesis of the ISREs and this may be essential when considering gene therapy in vivo. PMID- 10666453 TI - Tissue-specific chromatin structure of the phenobarbital-responsive unit and proximal promoter of CYP2B1/2 and modulation by phenobarbital. AB - Phenobarbital induction of transcription of CYP2B genes is mediated by an enhancer, termed a phenobarbital responsive unit (PBRU), approximately 2000 bp 5' of the transcription start site. To further delineate the mechanism of phenobarbital induction, protein binding in native chromatin and the nucleosomal structure of the PBRU and proximal promoter were examined in liver and kidney, in which the CYP2B1/2 genes are expressed and not expressed, respectively. Protein binding to the PBRU in kidney chromatin was not detected even though in vitro DNase I footprints were not detectably different with nuclear extracts from liver and kidney. Likewise, protein binding to regulatory motifs was not detected in the proximal promoter region in kidney chromatin. In liver chromatin, however, DNase I hypersensitivity and partial protection of the regulatory motifs from DNase I digestion or reaction with dimethyl sulfate was observed and phenobarbital treatment increased the hypersensitivity but only modestly affected protection. Low resolution Southern analysis of micrococcal nuclease-digested chromatin from untreated rats revealed micrococcal nuclease hypersensitive regions in the proximal promoter and PBRU regions in liver, but not in kidney. Phenobarbital treatment increased hyper-sensitivity in liver in both regions. Micrococcal nuclease hypersensitivity in the PBRU was largely restricted to a linker region between phased nucleosomes while in the proximal promoter hypersensitivity extended over approximately 200 bp suggesting disruption of a nucleosome in this region. These data indicate that in liver phenobarbital treatment substantially alters protein binding to regulatory motifs in the PBRU, while not greatly affecting such binding in the proximal promoter, and substantially alters chromatin structure in both regions, presumably as a result of chromatin modifying factors recruited to the PBRU. In the kidney, chromatin is probably in a closed conformation that prevents binding of regulatory factors. PMID- 10666454 TI - Polymorphisms in the large subunit of human RNA polymerase II as target for allele-specific inhibition. AB - The lack of specificity of cancer treatment causes damage to normal cells as well, which limits the therapeutic range. To circumvent this problem one would need to use an absolute difference between normal cells and cancer cells as therapeutic target. Such a difference exists in the genome of all individuals suffering from a tumor that is characterized by loss of genetic material [loss of heterozygosity (LOH)]. Due to LOH, the tumor is hemizygous for a number of genes, whereas the normal cells of the individual are heterozygous for these genes. Theoretically, polymorphic sites in these genes can be utilized to selectively target the cancer cells with an antisense oligonucleotide, provided that it can discriminate the alleles and inhibit gene expression. Furthermore, the targeted gene should be essential for cell survival, and 50% gene expression sufficient for the cell to survive. This will allow selective killing of cancer cells without concomitant toxicity to normal cells. As an initial step in the experimental test of this putative selective cancer cell therapy, we have developed a set of antisense phosphorothioate oligonucleotides which can discriminate the two alleles of a polymorphic site in the gene encoding the large subunit of RNA polymerase II. Our data show that the exact position of the antisense oligonucleotide on the mRNA is of essential importance for the oligo nucleotide to be an effective inhibitor of gene expression. Shifting the oligonucleotide position only a few bases along the mRNA sequence will completely abolish the inhibitory activity of the antisense oligonucleotide. Reducing the length of the oligonucleotides to 16 bases increases the allele specificity. This study shows that it is possible to design oligonucleotides that selectively target the matched allele, whereas the expression level of the mismatched allele, that differs by one nucleotide, is only slightly affected. PMID- 10666455 TI - Characterization of the E.coli poly(A) polymerase: nucleotide specificity, RNA binding affinities and RNA structure dependence. AB - Polyadenylation of RNA molecules in bacteria and chloroplasts has been implicated as part of the RNA degradation pathway. The polyadenylation reaction is performed in Escherichia coli mainly by the enzyme poly(A) polymerase I (PAP I). In order to understand the molecular mechanism of RNA poly-adenylation in bacteria, we characterized the biochemical properties of this reaction in vitro using the purified enzyme. Unlike the PAP from yeast nucleus, which is specific for ATP, E.coli PAP I can use all four nucleotide triphosphates as substrates for addition of long ribohomopolymers to RNA. PAP I displays a high binding activity to poly(U), poly(C) and poly(A) ribohomopolymers, but not to poly(G). The 3'-ends of most of the mRNA molecules in bacteria are characterized by a stem-loop structure. We show here that in vitro PAP I activity is inhibited by a stem-loop structure. A tail of two to six nucleo-tides located 3' to the stem-loop structure is sufficient to overcome this inhibition. These results suggest that the stem-loop structure located in most of the mRNA 3'-ends may function as an inhibitor of poly-adenylation and degradation of the corresponding RNA molecule. However, RNA 3'-ends produced by endonucleolytic cleavage by RNase E in single strand regions of mRNA molecules may serve as efficient substrates for polyadenylation that direct these molecules for rapid exonucleolytic degradation. PMID- 10666456 TI - Regulation of double-strand break-induced mammalian homologous recombination by UBL1, a RAD51-interacting protein. AB - Mammalian RAD51 protein plays essential roles in DNA homologous recombination, DNA repair and cell proliferation. RAD51 activities are regulated by its associated proteins. It was previously reported that a ubiquitin-like protein, UBL1, associates with RAD51 in the yeast two-hybrid system. One function of UBL1 is to covalently conjugate with target proteins and thus modify their function. In the present study we found that non-conjugated UBL1 forms a complex with RAD51 and RAD52 proteins in human cells. Overexpression of UBL1 down-regulates DNA double-strand break-induced homologous recombination in CHO cells and reduces cellular resistance to ionizing radiation in HT1080 cells. With or without overexpressed UBL1, most homologous recombination products arise by gene conversion. However, overexpression of UBL1 reduces the fraction of bidirectional gene conversion tracts. Overexpression of a mutant UBL1 that is incapable of being conjugated retains the ability to inhibit homologous recombination. These results suggest a regulatory role for UBL1 in homologous recombination. PMID- 10666457 TI - Understanding oligonucleotide-mediated inhibition of gene expression in Xenopus laevis oocytes. AB - Triplex-forming oligonucleotides (TFOs) modified with N,N-diethylethylenediamine can inhibit the expression of a reporter plasmid in Xenopus oocytes if the triplex is preformed prior to injection while unmodified oligonucleotides cannot. Here we show that merely forming a triplex in a reporter plasmid does not disrupt transcription, but when TFOs are targeted to sites within the transcribed region of a reporter gene then gene activity is inhibited. TFO-based inhibition did not lead to large scale degradation or mutation of the reporter plasmid, but dramatically lowered mRNA levels. Finally, we investigated the accessibility of a triplex target site on a reporter plasmid after injection into nuclei. We found that the site used for our previous studies was inaccessible to restriction endonuclease after injection into nuclei. This observation may explain why inhibition was dependent on forming the triplex before injection into oocytes. Based on the assumption that oligonucleotide association, like restriction enzyme access, was excluded by nucleosome formation, additional target sites were inserted so that all sites could not simultaneously be associated with the octamer core of a nucleosome. With multiple target sites prior association of the plasmid with nuclear proteins does not prevent oligonucleotide-mediated inhibition of gene activity. PMID- 10666458 TI - Recognition of 5-aminouracil (U(#)) in the central strand of a DNA triplex: orientation selective binding of different third strand bases. AB - A necessary feature of the natural base triads for triplex formation is the requirement of a purine (A or G) in the central position, since only these provide sets of two hydrogen bond donors/acceptors in the major groove of the double helix. Pyrimidine bases devoid of this feature have incompatible complementarity and lead to triplexes with lower stability. This paper demonstrates that 5-aminouracil (U#) (I), a pyrimidine nucleobase analogue of T in which 5-methyl is replaced by 5-amino group, with hydrogen bonding sites on both sides, is compatible in the central position of triplex triad X*U# x A, where X = A/G/C/T/2-aminopurine (AP), and * and x represent Hoogsteen and Watson Crick hydrogen bonding patterns respectively. A novel recognition selectivity based on the orientation (parallel/antiparallel) of the third strand purines A, G or AP with A in the parallel motif (A(p)*U# x A), and G/AP in the antiparallel motif (G(ap)/AP(ap)*U# x A) is observed. Similarly for pyrimidines in the third strand, C is accepted only in a parallel mode (C(p)*U(#) x A). Significantly, T is recognised in both parallel and antiparallel modes (T(p)/T(ap)*U(#) x A), with the antiparallel mode being stable compared to the parallel one. The 'U(#)' triplexes are also more stable than the corresponding control 'T' triplexes. The results expand the lexicon of triplex triads with a recognition motif consisting of pyrimidine in the central strand. PMID- 10666459 TI - Stereochemical control of DNA biosynthesis. AB - Stereochemical control of DNA biosynthesis was studied using several DNA synthesizing complexes containing, in each case, a single substitution of a 2' deoxy-D-nucleotide residue by an enantiomeric L-nucleotide residue in a DNA chain (either in the primer or in the template) as well as 2'-deoxy-L-ribonucleoside 5' triphosphates (L-dNTPs) as substrates. Three template-dependent DNA polymerases were tested, Escherichia coli DNA polymerase I Klenow fragment, Thermus aquaticus DNA polymerase and avian myeloblastosis virus reverse transcriptase, as well as template-independent calf-thymus terminal deoxynucleotidyl transferase. Very stringent control of stereoselectivity was demonstrated for template-dependent DNA polymerases, whereas terminal deoxynucleotidyl transferase was less selective. DNA polymerase I and reverse transcriptase catalyzed formation of dinucleoside 5',5'-tetraphosphates when L-dTTP was used as substrate. Comparison between models of template-primer complexes, modified or not by a single L nucleotide residue, revealed striking differences in their geometry. PMID- 10666460 TI - Specific bonding of puromycin to full-length protein at the C-terminus. AB - Puromycin, an analog of the 3' end of aminoacyl-tRNA, causes premature termination of translation by being linked non-specifically to growing polypeptide chains. Here we report the interesting phenomenon that puromycin acting as a non-inhibitor at very low concentration (e.g. 0.04 microM) can bond only to full-length protein at the C-terminus. This was proved by using a carboxypeptidase digestion assay of the products obtained by Escherichia coli cell-free translation of human tau 4 repeat (tau4R) mRNA in the presence of low concentrations of puromycin or its derivatives. The tau4R mRNA was modified to code for three C-terminal methionines, which were radioactively labeled, followed by a stop codon. The translation products could not be digested by carboxy peptidase if puromycin or a derivative was present at the C-terminus of full length tau4R. Puromycin and its derivatives at 0. 04-1.0 microM bonded to 7-21% of full-length tau4R, depending on the ability to act as acceptor substrates. Furthermore, the bonding efficiency of a puromycin derivative to tau4R was decreased by addition of release factors. These results suggest that puromycin and its derivatives at concentrations lower than those able to compete effectively with aminoacyl-tRNA can bond specifically to full-length protein at a stop codon. This specific bonding of puromycin to full-length protein should be useful for in vitro selection of proteins and for in vitro and in vivo C-terminal end protein labeling. PMID- 10666462 TI - DNA sequence analysis by hybridization with oligonucleotide microchips: MALDI mass spectrometry identification of 5mers contiguously stacked to microchip oligonucleotides. AB - Matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) has been applied to increase the informational output from DNA sequence analysis. It has been used to analyze DNA by hybridization with microarrays of gel-immobilized oligonucleotides extended with stacked 5mers. In model experiments, a 28 nt long DNA fragment was hybridized with 10 immobilized, overlapping 8mers. Then, in a second round of hybridization DNA-8mer duplexes were hybridized with a mixture of 10 5mers. The stability of the 5mer complex with DNA was increased to raise the melting temperature of the duplex by 10-15 degrees C as a result of stacking interaction with 8mers. Contiguous 13 bp duplexes containing an internal break were formed. MALDI MS identified one or, in some cases, two 5mers contiguously stacked to each DNA-8mer duplex formed on the microchip. Incorporating a mass label into 5mers optimized MALDI MS monitoring. This procedure enabled us to reconstitute the sequence of a model DNA fragment and identify polymorphic nucleotides. The application of MALDI MS identification of contiguously stacked 5mers to increase the length of DNA for sequence analysis is discussed. PMID- 10666461 TI - The catalytic subunit DNA-dependent protein kinase (DNA-PKcs) facilitates recovery from radiation-induced inhibition of DNA replication. AB - Exposure of cells to ionizing radiation inhibits DNA replication in a dose dependent manner. The dose response is biphasic and the initial steep component reflects inhibition of replicon initiation thought to be mediated by activation of the S-phase checkpoint. In mammalian cells, inhibition of replicon initiation requires the ataxia telagiectasia mutated ( ATM ) gene, a member of the phosphatidyl inositol kinase-like (PIKL) family of protein kinases. We studied the effect on replicon initiation of another member of the PI-3 family of protein kinases, the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) by measuring either total DNA synthesis, or size distribution of nascent DNA using alkaline sucrose gradient centrifugation. Exposure of human cells proficient in DNA-PKcs (HeLa or M059-K) to 10 Gy inhibited replicon initiation in a time dependent manner. Inhibition was at a maximum 1 h after irradiation and recovered at later times. Similar treatment of human cells deficient in DNA-PKcs (M059-J) inhibited replicon initiation to a similar level and with similar kinetics; however, no evidence for recovery, or only limited recovery, was observed for up to 8 h after irradiation. In addition a defect was observed in the maturation of nascent DNA. Similarly, a Chinese hamster cell line deficient in DNA-PKcs (irs 20) showed little evidence for recovery of DNA replication inhibition up to 6 h after irradiation, whereas the parental CHO cells showed significant recovery and an irs-20 derivative expressing the human DNA-PKcs complete recovery within 4 h. Normal kinetics of recovery were observed in xrs-5 cells, deficient in Ku80; in 180BR cells, deficient in DNA ligase IV; as well as XR-1 cells, deficient in XRCC4, an accessory factor of DNA ligase IV. Since all these cell lines share the DNA double strand break rejoining defect of M059-J and irs20 cells, the lack of recovery of DNA replication in the latter cells may not be attributed entirely to the prolonged presence of unrepaired DNA dsb. We propose that DNA-PKcs, in addition to its functions in the rejoining of DNA dsb and in DNA replication, also operates in a pathway that in normal cells facilitates recovery of DNA replication after irradiation. PMID- 10666463 TI - Identification of a novel 70 kDa protein that binds to the core promoter element and is essential for ribosomal DNA transcription. AB - Mammalian ribosomal RNA genes (rDNA) are transcribed by RNA polymerase I and at least two auxiliary factors, UBF and SL1/TFID/TIF-IB. It has also been reported that an additional factor(s) is required to reconstitute efficient initiation of rDNA transcription in vitro, depending upon the procedures of chromatographic separation. In an attempt to elucidate the molecular identity of such yet uncertain activities, we have developed agarose gel shift and UV cross-linking assays to detect proteins directly bound to the core promoter region of murine rDNA. With these techniques, we identified a 70 kDa protein (p70) in the flow through fraction of a phosphocellulose column (TFIA-fraction). Interestingly, the binding of p70 to the rDNA core promoter was observed only in the presence of the SL1-containing fraction. The probable human orthologue of p70 was also detected in HeLa cells. Consistent with the observation that p70 bound to the core promoter only in the presence of the TFIA- and SL1-fractions, alteration of DNase I footprint pattern over the core promoter element was demonstrated by cooperative action of the TFIA- and SL1-fractions. A reconstituted in vitro transcription assay with further purified p70 indicated that p70 was required for accurate initiation of rDNA transcription. These results indicate that the p70 identified recently by the current DNA-binding experiments represents a novel transcription factor in rDNA transcription. PMID- 10666464 TI - Evaluation and characterization of catabolite-responsive elements (cre) of Bacillus subtilis. AB - A global mechanism of catabolite repression of the genus Bacillus comprises negative regulation exerted through the binding of the CcpA protein to the catabolite-responsive elements (cres) of the target genes. We searched for cre sequences in the Bacillus subtilis genome using a query sequence, WTGNAANCGNWNNCW (N and W stand for any base and A or T, respectively), picking out 126 putative and known cre sequences. To examine their cre function, we integrated spac promoter (P spac )-cre-lacZ fusions into the amyE locus. Examination of catabolite repression of beta-galactosidase synthesis in the integrants led us to the following conclusions: (i) lower mismatching of cre sequences to the query sequence is required for their function; (ii) although cre sequences are partially palindromic, low mismatching in the same direction as that of transcription of the target genes is more critical for their function than that in the inverse direction; and (iii) yet, a more palindromic nature of cre sequences is desirable for a better function. Furthermore, the alignment of 22 cre s that function in vivo implicated a consensus sequence, WWTGNAARCGNWWWCAWW (R stands for G or A). Interestingly, in the case where cre sequences are located in the protein-coding regions of the target genes, their conserved bases are preferentially the third bases of codons where base degeneracy is allowed. PMID- 10666465 TI - Poly(A)-binding protein I of Leishmania: functional analysis and localisation in trypanosomatid parasites. AB - Regulation of gene expression in trypanosomatid parasites is predominantly post transcriptional. Primary transcripts are trans-spliced and polyadenylated to generate mature mRNAs and transcript stability is a major factor controlling stage-specific gene expression. Degenerate PCR has been used to clone the gene encoding the Leishmania homologue of poly(A)-binding protein (Lm PAB1), as an approach to the identification of trans-acting factors involved in this atypical mode of eukaryotic gene expression. lmpab1 is a single copy gene encoding a 63 kDa protein which shares major structural features but only 35-40% amino acid identity with other PAB1 sequences, including those of other trypanosomatids. Lm PAB1 is expressed at constant levels during parasite differentiation and is phosphorylated in vivo. It is localised predominantly in the cytoplasm but inhibition of transcription with actinomycin D also reveals diffuse localisation in the nucleus. Lm PAB1 binds poly(A) with high specificity and affinity but fails to complement a null mutation in Saccharomyces cerevisiae. These properties are indicative of functional divergence in vivo. PMID- 10666466 TI - Evidence for evolutionarily conserved secondary structure in the H19 tumor suppressor RNA. AB - The molecular basis for function of the mammalian H19 as a tumor suppressor is poorly understood. Large, conserved open reading frames (ORFs) are absent from both the human and mouse cDNAs, suggesting that it may act as an RNA. Contradicting earlier reports, however, recent studies have shown that the H19 transcript exists in polysomal form and is likely translated. To distinguish between possible functional roles for the gene product, we have characterized the sequence requirements for H19-mediated in vitro suppression of tumor cell clonogenicity and analyzed the sequence of the gene cloned from a range of mammals. A cDNA version of the human gene, lacking the unusually short introns characteristic of imprinted genes, is as effective as a genomic copy in blocking anchorage-independent growth by G401 cells. The first 710 nucleotides of the gene can be deleted with no effect on in vitro activity. Further truncations from either the 5'- or 3'-end, however, cause a loss of suppression of clonogenicity. Using conserved sequences within the H19 gene as PCR primers, genomic DNA fragments were amplified from a range of mammalian species that span the functional domain defined by deletion analysis. Sequences from cat, lynx, elephant, gopher and orangutan complement the previous database of sequences from human, mouse, rat and rabbit. Hypothetical translation of the resulting sequences shows an absence of conserved ORFs of any size. Free energy and covariational analysis of the RNA sequences was used to identify potential helical pairings within the H19 transcript. A set of 16 helices are supported by covariation (i.e. conservation of base pairing potential in the absence of primary sequence conservation). The predicted RNA pairings consist largely of local hairpins but also include several long range interactions that bridge the 5'- and 3'-ends of the functional domain. Given the evolutionary conservation of structure at the RNA level and the absence of conservation at the protein level, we presume that the functional product of the H19 gene is a structured RNA. PMID- 10666467 TI - A highly ordered structure in V(D)J recombination cleavage complexes is facilitated by HMG1. AB - Central to understanding the process of V(D)J recombination is appreciation of the protein-DNA complex which assembles on the recombination signal sequences (RSS). In addition to RAG1 and RAG2, the protein HMG1 is known to stimulate the efficiency of the cleavage reaction. Using electrophoretic mobility shift analysis we show that HMG1 stimulates the in vitro assembly of a stable complex with the RAG proteins on each RSS. We use UV crosslinking studies of this complex with azido-phenacyl derivatized probes to map the contact sites between the RAG proteins, HMG1 derivatives and the RSS. We find that the RAG proteins make contacts at the nonamer, heptamer and adjacent coding region. The HMG1 protein by itself appears to localize at the 3' side of the nonamer, but a cooperative complex with the RAG proteins is positioned at the 3' side of the heptamer and adjacent spacer in the 12RSS. In the complex with RAG proteins, HMG1 is positioned primarily in the spacer of the 23RSS. We suggest that bends introduced into these DNA substrates at specific locations by the RAG proteins and HMG1 may help distinguish the 12RSS from the 23RSS and may therefore play an important role in the coordinated reaction. PMID- 10666468 TI - Protein sequences conserved in prokaryotic aminoacyl-tRNA synthetases are important for the activity of the processivity factor of human mitochondrial DNA polymerase. AB - Previous studies have shown that the small subunit of Xenopus DNA polymerase gamma (pol gammaB) acts as a processivity factor to stimulate the 140 kDa catalytic subunit of human DNA polymerase gamma. A putative human pol gammaB initially identified by analysis of DNA sequence had not been shown to be functional, and appeared to be an incomplete clone. In this paper, we report the cloning of full-length human and mouse pol gammaB. Both human and mouse pol gammaB proteins were expressed in their mature forms, without their apparent mitochondrial localization signals, and shown to stimulate processivity of the recombinant catalytic subunit of human pol gammaA. Deletion analysis of human pol gammaB indicated that blocks of sequence conserved with prokaryotic class II aminoacyl-tRNA synthetases are necessary for activity and inter-action with human pol gammaA. Purification of DNA pol gamma from HeLa cells indicated that both proteins are associated in vivo. PMID- 10666469 TI - Purification and characterization of the DNA cleavage and recognition site of I ScaI mitochondrial group I intron encoded endonuclease produced in Escherichia coli. AB - The second intron in the mitochondrial cytb gene of Saccharomyces capensis, belonging to group I, encodes a 280 amino acid protein containing two LAGLIDADG motifs. Genetic and molecular studies have previously shown that this protein has a dual function in the wild-type strain. It acts as a specific homing endonuclease I- Sca I promoting intron mobility and as a maturase promoting intron splicing. Here we describe the synthesis of a universal code equivalent to the mitochondrial sequence coding for this protein and the in vitro characterization of I- Sca I endonuclease activity, using a truncated mutant form of the protein p28bi2 produced in Escherichia coli. We have also determined the cleavage pattern as well as the recognition site of p28bi2. It was found that p28bi2 generates a double-strand cleavage downstream from the intron insertion site with 4 nt long 3'-overhangs. Mutational analysis of the DNA target site shows that p28bi2 recognizes a 16-19 bp sequence from positions -11 to +8 with respect to the intron insertion site. PMID- 10666470 TI - Structures of m-iodo Hoechst-DNA complexes in crystals with reduced solvent content: implications for minor groove binder drug design. AB - The DNA photosensitisers m-iodo Hoechst and m-iodo, p-methoxy Hoechst have been co-crystallised with the oligonucleotide d(CGCGAATTCGCG)(2)and their crystal structures determined. The crystals were then subjected to slow dehydration, which reduced their solvent contents from 40 (normal) to 30 (partially dehydrated) and then 20% (fully dehydrated) and caused a reduction in cell volume from 68,000 to 60,000 then 51,000 A(3). The dehydration resulted in a dramatic enhancement of diffraction resolution from approximately 2.6 to beyond 1.5 A. Crystal structures have also been determined for the partially and fully dehydrated states. The fully dehydrated crystals consist of an infinite polymeric network, in which neighbouring dodecamer duplexes are crosslinked through phosphate oxygens via direct bonding to bridging magnesium cations. This unique three-dimensional structure for DNA is described in detail in the following companion paper. The present paper details evidence from the sequence of crystal structures that the DNA is able to breathe locally, allowing the ligand to leave the minor groove, re-orient in the surrounding solvent medium and then re-enter the groove in a different orientation and location. The rearrangement of the minor groove binding ligands during the dehydration process mimics the binding behaviour of these ligands in solution and in vivo. We also present details of the DNA-ligand interactions that are consistent with a hydrogen atom ion mechanism for photocleavage of DNA. PMID- 10666471 TI - Intermolecular interactions and water structure in a condensed phase B-DNA crystal. AB - By controlled dehydration, the unit cells of dodecamer DNA-drug crystals have been shrunk from 68,000 (normal state) to 60,000 (partially dehydrated intermediate state) to 51,000 A(3) (fully dehydrated state), beyond which no further solvent loss occurs. The total solvent content in the normal crystals is approximately 40% by volume, reducing to approximately 20% in the fully dehydrated phase. The 25% reduction in cell volume induced a dramatic enhancement in the resolution of the X-ray diffraction data (from 2. 6 to beyond 1.5 A). We have determined the structures of the normal, partially dehydrated and fully dehydrated crystals. Details of the ligand binding have been presented in the preceding article. The present paper describes the unique features of the structure of the fully dehydrated phase. This structure was refined with 9,015 unique observed reflections to R = 14.9%, making it one of the most reliable models of B -form DNA available. The crystals exist as infinite polymeric networks, in which neighbouring dodecamer duplexes are crosslinked through phosphate oxygens via direct bonding to magnesium cations. The DNA is packed so tightly that there is essentially only a single layer of solvent between adjacent molecules. The details of the crystal packing, magnesium bridging, DNA hydration and DNA conformation are described and compared with other experimental evidence related to DNA condensation. PMID- 10666472 TI - RNA-binding properties of the mitochondrial Y-box protein RBP16. AB - We have previously identified a mitochondrial Y-box protein in Trypanosoma brucei that we designated RBP16. The predicted RBP16 amino acid sequence revealed the presence of a cold-shock domain at its N-terminus and a glycine- and arginine rich C-terminus reminiscent of an RGG RNA-binding motif. Since RBP16 is capable of interacting with different guide RNAs (gRNAs) in vitro and in vivo primarily via the oligo(U) tail, as well as with ribosomal RNAs, possible functions of RBP16 may be in kinetoplastid RNA editing and/or translation. Herein, we report experiments that further define the RNA-binding properties of RBP16. RBP16 forms a single stable complex with the gRNA gA6[14] at low protein concentration, while at higher protein concentration two stable complexes that possibly represent two different conformations are observed. Both complexes are stable at relatively high salt and moderate heparin concentrations indicating that the binding of RBP16 to gA6[14] does not rely primarily on ionic interactions. Phenylglyoxal treatment of the protein indicates that arginine residues are important in RNA binding. The minimal length of RNA sequence necessary for the binding of RBP16 was assessed by gel retardation and UV cross-linking competition assays using oligo(U) ribonucleotides of varying lengths (4-40 nt). Although RBP16 can bind to oligonucleotides as small as U(4), its affinity increases with the length of the oligo(U) ribonucleotide, with a dramatic increase in binding efficiency observed when the length is increased to 10 nt. Gel retardation assays employing T.brucei mRNAs demonstrated that, although it acts as a major binding determinant, a 3' U tail is not an absolute requirement for efficient RBP16-RNA binding. Experiments with oligonucleotides containing U stretches embedded at different positions in oligo(dC) indicated that high-affinity binding requires both a uridine stretch, as well as 5' and 3' non-specific sequences. These results suggest a model for the molecular interactions involved in RBP16-RNA binding. PMID- 10666473 TI - Terminal deoxynucleotidyl transferase catalyzes the reaction of DNA phosphorylation. AB - The reaction of phosphorylation and phosphonylation of an oligodeoxynucleotide 3' terminal hydroxyl (oligodeoxynucleotidyl kinase activity) catalyzed by calf thymus terminal deoxynucleotidyl transferase (TDT) was found. Triphosphates modified at Palpha-, Palpha,gamma- or Palpha,beta,gamma-residues served as low molecular weight substrates. The reaction was TDT specific; human DNA polymerasesalphaandbeta, as well as AMV reverse transcriptase did not catalyze it. The donor activity of modified triphosphates or triphosphonates depended on their structure and was increased with an increase in their hydrophobicity. The substrate activity of some modified triphosphates was up to one order of magnitude higher than that of ddTTP. PMID- 10666474 TI - A novel procedure for efficient genotyping of single nucleotide polymorphisms. AB - Due to the surge in interest in using single nucleotide polymorphisms (SNPs) for genotyping a facile and affordable method for this is an absolute necessity. Here we introduce a procedure that combines an easily automatable single tube sample preparation with an efficient high throughput mass spectrometric analysis technique. Known point mutations or single nucleotide polymorphisms are easily analysed by this procedure. It starts with PCR amplification of a short stretch of genomic DNA, for example an exon of a gene containing a SNP. By shrimp alkaline phosphatase digest residual dNTPs are destroyed. Allele-specific products are generated using a special primer, a conditioned set of alpha-S-dNTPs and alpha-S-ddNTPs and a fresh DNA polymerase in a primer extension reaction. Unmodified DNA is removed by 5'-phospho-diesterase digestion and the modified products are alkylated to increase the detection sensitivity in the mass spectrometric analysis. All steps of the preparation are simple additions of solutions and incubations. The procedure operates at the lowest practical sample volumes and in contrast to other genotyping protocols with mass spectrometric detection requires no purification. This reduces the cost and makes it easy to implement. Here it is demonstrated in a version using positive ion detection on described mutations in exon 17 of the amyloid precursor protein gene and in a version using negative ion detection on three SNPs of the granulocyte-macrophage colony stimulating factor gene. Preparation and analysis of SNPs is shown separately and simultaneously, thus demonstrating the multiplexibility of this genotyping procedure. The preparation protocol for genotyping is adapted to the conditions used for the SNP discovery method by denaturing HPLC, thus demonstrating a facile link between protocols for SNP discovery and SNP genotyping. Results corresponded unanimously with the control sequencing. The procedure is useful for high throughput genotyping as it is required for gene identification and pharmacogenomics where large numbers of DNA samples have to be analysed. We have named this procedure the 'GOOD Assay' for SNP analysis. PMID- 10666475 TI - Functional coexpression of serine protein kinase SRPK1 and its substrate ASF/SF2 in Escherichia coli. AB - Mammalian proteins expressed in Escherichia coli are used in a variety of applications. A major drawback in producing eukaryotic proteins in E.coli is that the bacteria lack most eukaryotic post-translational modification systems, including serine/threonine protein kinase(s). Here we show that a eukaryotic protein can be phosphorylated in E.coli by simultaneous expression of a mammalian protein kinase and its substrate. We show that in bacteria expressing SRPK1, ASF/SF2 becomes phosphorylated to a degree resembling native ASF/SF2 present in interphase HeLa cell nuclei. The E.coli phosphorylated ASF/SF2 is functional in splicing and, contrary to the unphosphorylated protein, soluble under native conditions. PMID- 10666476 TI - How cells use proteolysis to control their growth. PMID- 10666478 TI - Macroscopic spectral imaging and gene expression analysis of the early stages of melanoma. AB - BACKGROUND: The stages of melanocytic progression are defined as atypical (dysplastic) nevus, melanoma in situ, melanoma in the radial growth phase (RGP), melanoma in the vertical growth phase (VGP), and melanoma in the metastatic growth phase (MGP). Melanoma in situ and RGP melanoma often develop in contiguous association with atypical nevi. This frequently poses a problem with respect to their early detection. Furthermore, unlike cells obtained from VGP and MGP melanomas, cells derived from melanoma in situ and RGP melanoma do not proliferate in vitro. Thus, compared to the late stages of the disease, less information is available regarding genes expressed in the early stages. MATERIALS AND METHODS: To determine whether spectral imaging, a recently developed optical imaging technique, can detect melanoma in situ and RGP melanoma arising in melanoma precursor lesions, atypical nevi in patients with a clinical history of melanoma were subjected to noninvasive macroscopic spectral imaging. To determine at what stage in the progression pathway of melanoma genes having important biological functions in VGP and MGP melanomas are activated and expressed, lesions of melanoma in situ were analyzed by immunohistochemistry and in situ hybridization for expression of some of these known molecular and immunologic markers. RESULTS: The present study demonstrates the capability of noninvasive spectral imaging to detect melanoma in situ and RGP melanoma that arise in contiguous association with atypical nevi. Furthermore, the study provides evidence that genes and antigens expressed in VGP and MGP melanoma are also expressed in melanoma in situ. CONCLUSIONS: Because of the dark and variegated pigmentation of atypical nevi, melanoma in situ and RGP melanoma that arise in these melanoma precursor lesions are often difficult to recognize and thus frequently go unnoticed. The application of new optical screening techniques for early detection of melanoma and the identification of genes expressed in the early stages of melanoma development are two important avenues in the pursuit of melanoma prevention. The investigations presented here document that macroscopic spectral imaging has the potential to detect melanoma in its early stage of development and that genes essential for the proliferation and cell adhesion of VGP and MGP melanoma are already expressed in melanoma in situ. PMID- 10666479 TI - CXC and CC chemokine receptors on coronary and brain endothelia. AB - BACKGROUND: Chemokine receptors on leukocytes play a key role in inflammation and HIV-1 infection. Chemokine receptors on endothelia may serve an important role in HIV-1 tissue invasion and angiogenesis. MATERIALS AND METHODS: The expression of chemokine receptors in human brain microvascular endothelial cells (BMVEC) and coronary artery endothelial cells (CAEC) in vitro and cryostat sections of the heart tissue was determined by light and confocal microscopy and flow cytometry with monoclonal antibodies. Chemotaxis of endothelia by CC chemokines was evaluated in a transmigration assay. RESULTS: In BMVEC, the chemokine receptors CCR3 and CXCR4 showed the strongest expression. CXCR4 was localized by confocal microscopy to both the cytoplasm and the plasma membrane of BMVEC. In CAEC, CXCR4 demonstrated a strong expression with predominantly periplasmic localization. CCR5 expression was detected both in BMVEC and CAEC but at a lower level. Human umbilical cord endothelial cells (HUVEC) expressed strongly CXCR4 but only weakly CCR3 and CCR5. Two additional CC chemokines, CCR2A and CCR4, were detected in BMVEC and CAEC by immunostaining. Immunocytochemistry of the heart tissues with monoclonal antibodies revealed a high expression of CXCR4 and CCR2A and a low expression of CCR3 and CCR5 on coronary vessel endothelia. Coronary endothelia showed in vitro a strong chemotactic response to the CC chemokines RANTES, MIP 1alpha, and MIP-1beta. CONCLUSIONS: The endothelia isolated from the brain display strongly both the CCR3 and CXCR4 HIV-1 coreceptors, whereas the coronary endothelia express strongly only the CXCR4 coreceptor. CCR5 is expressed at a lower level in both endothelia. The differential display of CCR3 on the brain and coronary endothelia could be significant with respect to the differential susceptibility of the heart and the brain to HIV-1 invasion. In addition, CCR2A is strongly expressed in the heart endothelium. All of the above chemokine receptors could play a role in endothelial migration and repair. PMID- 10666480 TI - Twenty novel mutations in the alpha-galactosidase A gene causing Fabry disease. AB - BACKGROUND: Fabry disease, an X-linked inborn error of glycosphingolipid catabolism, results from the deficient activity of the lysosomal exoglycohydrolase alpha-galactosidase A (EC 3.2.1.22; alpha-Gal A). The nature of the molecular lesions in the alpha-Gal A gene in 30 unrelated families was determined to provide precise heterozygote detection, prenatal diagnosis, and define genotype-phenotype correlations. MATERIALS AND METHODS: Genomic DNA was isolated from affected males and/or carrier females from 30 unrelated families with Fabry disease. The entire alpha-Gal A coding region and flanking intronic sequences were analyzed by PCR amplification and automated sequencing. RESULTS: Twenty new mutations were identified, each in a single family: C142R, G183D, S235C, W236L, D244H, P259L, M267I, I289F, Q321E, C378Y, C52X, W277X, IVS4(+4), IVS6(+2), IVS6(-1), 35del13, 256del1, 892ins1, 1176del4, and 1188del1. In the remaining 10 unrelated Fabry families, 9 previously reported mutations were detected: M42V, R112C, S148R, D165V, N215S (in 2 families), Q99X, C142X, R227X, and 1072del3. Haplotype analysis using markers closely flanking the alpha-Gal A gene indicated that the two patients with the N215S lesion were unrelated. The IVS4(+4) mutation was a rare intronic splice site mutation that causes Fabry disease. CONCLUSIONS: These studies further define the heterogeneity of mutations in the alpha-Gal A gene causing Fabry disease, permit precise heterozygote detection and prenatal diagnosis, and help delineate phenotype-genotype correlations in this disease. 100 days and 12/14 16.2 +/- 1.7 days; p < 0.001). The analysis of graft infiltrating cells on day 5 after transplantation showed a significant decrease in the number of graft infiltrating cells in RDP1258-treated recipients compared to untreated ones (14.8 vs. 32.7%; p < 0.01). In addition, grafts from peptide treated animals showed significantly decreased expression of TNF-alpha mRNA and increased levels of iNOS mRNA. Our results are consistent with the recent observation that up-regulation of HO-1 results in the inhibition of several immune effector functions. Modulation of HO-1 activity may enable the development of novel immunomodulatory strategies in humans. PMID- 10666489 TI - Genetic variation and covariation of parathyroid hormone levels and bone density in the human population. AB - The present study was an attempt to evaluate the relative importance of familial/genetic factors in interindividual variation of plasma concentrations of parathyroid hormone (PTH) and bone mineral density (BMD). We also examined to what extent common genetic and environmental factors may be involved in covariation between the hormone concentrations and BMD levels. Ninety-five nuclear pedigrees (consisting of 187 males and 168 females, aged 18-91 and 18-86 years old, respectively), from several small villages in the Chuvasha Autonomy, Russia, were assessed for PTH, sex hormones, and BMD. PTH plasma levels were measured in duplicate by immunoradiometric assay using an N-tact PTH SP kit. Standard roentgenography was done from the second and third phalanges of the middle finger on both hands for assessment of compact and cancellous bone BMD separately. The present study clearly confirmed the results of the previous genetic analyses of BMD which indicated that between 47% and 60% of the total variance of BMD, adjusted for sex and age effects, were attributable to genetic factors. Genetic factors also contributed significantly to interindividual variation of PTH. Constraining these additive genetic effects to zero dramatically increased the likelihood ratio (P < 0.001), indicating that at least 30% of the hormone plasma variation was attributable to genetic sources. The results of bivariate decomposition analysis were not clear cut. Two types of bivariate analyses showed that PTH-BMD genetic correlations according to sex and between the opposite sexes were consistently negative, but only marginally significant. PMID- 10666490 TI - Bone mineral density and muscle strength in young men with mental retardation (with and without Down syndrome). AB - The objective of this study was to compare the bone mineral density (BMD) of men with Down syndrome (DS) to otherwise mentally retarded (MR) men and to investigate whether leg muscle strength of these patients is related to BMD. Two groups with MR (with and without DS) participated in the study, having met the following criteria: similar age, moderate to mild mental retardation, Tanner stage V of sexual development, similar age of beginning to walk, and equal motor activities. The DS group consisted of 8 men 23.9 +/- 4.2 years, and the MR group without DS consisted of 8 men 23.5 +/- 3.6 years. The two groups were compared with 10 sedentary students of the same age range (25.9 +/- 2.9 years) attending our University. The BMD of the 2(nd) to 4(th) lumbar vertebrae was measured in the PA projection and the mean density was expressed as g/cm(2). The isokinetic muscle strength of the right quadriceps femoris and hamstrings muscles was measured on a Cybex II isokinetic dynamometer. The value measured was peak torque at angular velocities at 60, 120, and 300 degrees.sec(-1). The results showed that BMD in DS individuals versus young adults (reference group of the scanner) was lower at the 26% level (T-score - 2.66 +/- 0.29) and significantly lower (P = 0.002) than that of the MR group. Significantly different muscle strength was observed between the DS and non-DS MR group (in quadriceps at 300 degrees.s(-1): P < 0.01, at 120 and 60 degrees. s(-1): P < 0.05; in hamstrings at 300 degrees.s( 1): P < 0.05). Higher differences in muscle strength were found between MR and control men, but no significant difference existed in BMD between them. Bivariate correlation showed that quadriceps strength significantly predicted the BMD in the DS patients. Active lifestyle and increased physical exercise to improve muscular strength should be instituted to avoid the development of osteoporosis in DS patients. PMID- 10666491 TI - Association of bone mineral density with polymorphism of the human calcium sensing receptor locus. AB - A strong correlation between bone mass and genetic factors has been shown in twins and family studies. Some of the genes involved would regulate bone metabolism, bone formation, and resorption, all processes that determine bone mass. One candidate genes, calcium-sensing receptor (CASR) in the parathyroid gland, regulates calcium homeostasis by sensing decreases in extracellular calcium level and effecting an increase in secretion of parathyroid hormone (PTH) and calcium (Ca) reabsorption in the kidney. We have investigated a possible association between the CA-repeat polymorphism at the human CASR gene locus and the bone mineral density (BMD) of radial bone in 472 postmenopausal Japanese women. Genotypes were classified into nine groups according to the number of CA repeats present, from 20 to 12. BMD was expressed as the adjusted BMD, which was the body mass index (BMI), and age-adjusted average BMD. The 247 women who had an A3 allele (228 bp, containing 18 repeats of CA) had significantly lower adjusted BMD (mean +/- SD: 0.303 +/- 0.059 versus 0.316 +/- 0.063 g/cm(2); P = 0.0308) than the participants (n = 201) who did not carry an allele of that size. This result suggests that genetic variation at the CASR gene locus is associated with some determinants for BMD in postmenopausal women. PMID- 10666492 TI - Vitamin D and estrogen receptor polymorphisms and bone mineral changes in postpartum women. AB - BsmI restriction fragment length polymorphism (RFLP) of the vitamin D receptor (VDR) gene and PvuII RFLPs of the estrogen receptor (ER) gene and their relation to changes in areal bone mineral density (BMD) were examined in 43 healthy postpartum Finnish women aged 31.3 (SD 4.7) years. BMD was measured by dual energy X-ray absorptiometry at lumbar spine, right femoral neck, and dominant distal radius immediately after delivery, 1 month after resumption of menses, and 1 year thereafter. The RFLPs were represented as Bb (BsmI) and Pp (PvuII), the capital letters denoting the absence of and the small letters the presence of the restriction sites. The frequency of VDR alleles was as follows: bb (20.9%), Bb (60.5%), and BB (18.6%), and that of ER alleles was pp (39.5%), Pp (51.2%), and PP (9.3%). Altogether, BMD decreased significantly during postpartum amenorrhea at all sites [the mean bone loss ranging from -1.2 (SD 3.6)% at the distal radius to -3.7 (2.9)% at the femoral neck], and increased after resumption of menses [the 1-year follow-up BMD values ranging from -1.0 (2.4)% at the femoral neck to +3.3 (4.0)% at the lumbar spine as compared with baseline]. No obvious genotype related differences were found between these changes. These results suggest that the BsmI and PvuII polymorphisms may not have substantial influence on BMD changes postpartum. PMID- 10666493 TI - Association of methylenetetrahydrofolate reductase (MTHFR) polymorphism with bone mineral density in postmenopausal Japanese women. AB - The pathogenesis of osteoporosis is controlled by genetic and environmental factors. Considering the high prevalence of osteoporosis in homocystinuria, abnormal homocysteine metabolism would contribute to the pathogenesis of osteoporosis. It is known that the polymorphism of methylenetetrahydrofolate reductase (MTHFR), the enzyme catalyzing the reduction of 5, 10 methylenetetrahydrofolate to 5-methyltetrahydrofolate, correlates with hyperhomocysteinemia. In this study, we examined the association of this polymorphism with bone mineral density (BMD). BMD was measured by dual-energy X ray absorptiometry (DXA) in 307 postmenopausal women. MTHFR A/V polymorphism was analyzed using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). We compared BMD, clinical characteristics, and bone metabolic markers among MTHFR groups (AA, AV, VV). The groups did not differ in terms of baseline data. The values of lumbar spine BMD and total body BMD were as follows: lumbar spine: AA, 0.91 +/- 0.18, AV, 0.88 +/- 0.16, VV, 0.84 +/- 0.14 g/cm(2); total body: AA, 0.97 +/- 0.11, AV, 0.96 +/- 0.11, VV, 0.93 +/- 0.09 g/cm(2). In the VV genotype, lumbar spine BMD values were significantly lower than those of the women with the AA genotype (P = 0.016) and total body BMD was significantly lower than those of the women with AA genotype (P = 0.03) and AV genotype (P = 0.04). This is the first report that suggests that the VV genotype of MTHFR is one of the genetic risk factors for low BMD. PMID- 10666494 TI - Acute alteration in bone mineral density and biochemical markers for bone metabolism in nephrotic patients receiving high-dose glucocorticoid and one-cycle etidronate therapy. AB - It is widely known that glucocorticoids induce and accelerate osteoporosis. High dose glucocorticoids are administrated daily to patients in the acute phase of nephrotic syndrome. It could be inferred that high-dose glucocorticoids rapidly decrease patients' basal bone mineral density (BMD) and this accelerates the natural progress of osteoporosis associated with aging or menopause. Nine nephrotic patients (male/female: 5/4) without previous prednisolone administration were chosen to measure BMD and the level of the markers for bone turnover before and after treatment for 3 months (total prednisolone administration: 4.5 +/- 0.0 g). Twenty-three patients under remission with prednisolone administration (male/female: 14/9) were included in the long-term treatment group. Patients in this group whose %YAM in the lateral lumbar spine was less than 89% were classified into a low BMD group (n = 10, male/female: 3/7). They were administered etidronate disodium at 200 mg/day for 14 days. BMD and % of young adult mean (YAM) in the lumbar spine (L2-L4 in lateral objection) and other regions were measured by dual-energy X-ray absorptiometry. As markers of bone metabolism, the urinary level of deoxypyridinoline (Dpd) was determined to evaluate osteogenesis, and serum osteocalcin was measured to evaluate bone resorption. BMD of the lumbar spine significantly decreased in the 3-month treatment group (752 +/- 96 mg/cm(2), 7 +/- 4% reduction) compared with the pretreatment group (810 +/- 85 mg/cm(2)). BMD in the long-term treatment group decreased continuously (683 +/- 135 mg/cm(2)). No significant differences were noted in other measurement sites. BMD in the etidronate treatment group increased significantly (597 +/- 55 mg/cm(2)) compared with the pretreatment group (549 +/- 76 mg/cm(2)). Etidronate did not change BMD at the sites with a normal BMD. Among the biochemical markers (BM) examined, the urinary level of Dpd (nMol/liter. Cr) significantly increased in the 3-month treatment group (8.6 +/- 5.1 nMol/liter.Cr) compared with the pretreatment group (5.8 +/- 2.0 nMol/liter. Cr). No significant differences were seen in the BMs measured in the long-term treatment group. The urinary Dpd level of the etidronate treatment group decreased (3.9 +/- 1.4 nMol/liter. Cr) compared with the pretreatment group. These data indicate that etidronate could improve the accelerated bone resorption. In conclusion, high-dose glucocorticoid therapy causes rapid bone resorption and accelerates the natural progress of osteoporosis associated with aging or menopause. Etidronate administration prevents the progress of osteoporosis in nephrotic patients. Preventive treatment should be performed when the estimated BMD in 3 months falls below the baseline by more than 7 +/- 4%, reaching the therapeutic range. PMID- 10666495 TI - Estrogen deficiency is a potential cause for osteopenia in adult male patients with Noonan's syndrome. AB - Osteopenia is frequently observed in patients with Turner's syndrome. By contrast, there is no report concerning bone metabolism in patients with Noonan's syndrome which comprises Turner's phenotypic characteristics without any sex chromosome abnormalities. In the present investigation, we determined bone mineral density (BMD) as well as serum and urine indices of bone turnover in two male patients with Noonan's syndrome. Both patients showed remarkably decreased BMD, measured at two sites on the lumbar spine (L2-L4) and the distal end of the radius using dual energy X-ray absorptiometry (DXA). Urinary pyridinoline (PYD) and deoxypyridinoline (DPD) concentrations were significantly elevated in both patients, and serum osteocalcin and carboxyterminal propeptide of type I procollagen (PICP) concentrations were elevated in one patient. Surprisingly, both patients had a low level of serum 17beta-estradiol compared with control males, whereas they had normal levels of serum testosterone and dihydrotestosterone. Conjugated estrogens (Premarin 0.625 mg/day) were continued to be administered to these patients, followed up for 12 months. Urinary PYD and DPD concentrations gradually decreased, followed by an increase in their BMD. This is the first report that male patients with Noonan's syndrome showed osteopenia associated with increased bone resorption. Our data indicate that hypoestrogenism plays a potentially significant role in the abnormal bone metabolism in these patients. PMID- 10666496 TI - Acellular mineral deposition in collagen-based biomaterials incubated in cell culture media. AB - Rapid developments in tissue engineering have renewed interest in biodegradable three-dimensional structures such as collagen-based biomaterials. Collagen matrices seeded in vitro with fibroblasts, osteoblasts, and chondrocytes can form tissues resembling skin, bone, and cartilage that could be used as functional substitutes for damaged tissues. Collagen is associated with both dystrophic calcification of collagenous implants and bone mineralization. We report here the calcification properties of collagen sponges incubated in cell-free media. Mineral deposited in sponges was identified by X-ray and electron diffraction, Fourier transform infrared spectroscopy, and the molar ratio of calcium:phosphorus (Ca:P) as a poorly crystalline apatite similar to bone. The degree of calcification increased with length of incubation and the Ca and P content of the media, with 10-15% Ca (dry weight) after 21 days' incubation in media containing 1.6-3 mM Ca and a Ca x P molar product of 2-3 mM(2), but only 2% Ca after incubation in medium with 1.33 mM Ca and a 1.7 mM(2) Ca x P molar product. Mineral deposition was completely inhibited in sponges that were washed extensively and initially contained less than 0.01% P. Readdition of phosphate in these sponges and subsequent freeze drying and sterilization restore their mineralization capacity, suggesting that collagen per se cannot initiate calcification and that the inorganic phosphate content associated with the collagen preparation process is in the solid state a potential nucleator. Addition of chondroitin 4-sulfate to the sponges partially or totally inhibited mineral deposition, even though 80-90% of the compound was released within 24 hours. These results indicate that acellular calcification of collagen-based biomaterials can occur under the culture conditions currently used in tissue engineering. PMID- 10666497 TI - A novel method to isolate odontoblasts from rat incisor. AB - Historically, odontoblasts have been isolated from rat incisor using a surgical curette to separate these cells from the dentin. Isolation of odontoblasts using this approach typically resulted in cells with membrane properties that made the application of patch-clamp electrophysiological techniques prohibitive. The studies here describe a new procedure for isolating mature odontoblasts from adult rat incisor to obtain enriched populations of intact, viable odontoblasts that can be readily studied using patch-clamp methodologies. Identification of isolated cells as odontoblasts was confirmed using in situ mRNA hybridization for expression of dentin sialoprotein, osteocalcin, bone sialoprotein, and type I collagen, and calcium flux was monitored in these cells by means of fura-2 microfluorometry. We suggest that either single odontoblasts or clusters of these cells isolated by this new method would be an ideal preparation for the study of odontoblast properties using electrophysiological techniques, in situ hybridization and/or microfluorometry. PMID- 10666498 TI - Prostaglandin E(2) (PGE(2)) induces the c-fos and c-jun expressions via the EP(1) subtype of PGE receptor in mouse osteoblastic MC3T3-E1 cells. AB - This study examined which subtype(s) of PGE receptors is involved in the induction of c-fos and c-jun by PGE(2) in MC3T3-E1 cells. We also investigated the possibility that the induction of these genes is involved in the growth and differentiation of this cell line. PGE(2) dose-dependently induced c-fos and c jun mRNA expressions in MC3T3-E1 cells. Of the PGE analogs, 17-phenyl-omega trinor PGE(2) (EP(1) agonist) and sulprostone (EP(1)/EP(3) agonist) were far more potent than butaprost (EP(2) agonist) and 11-deoxy PGE(1) (EP(2)/EP(4) agonist) in inducing c-fos and c-jun mRNA expressions. Since MC3T3-E1 cells do not express the EP(3) subtype, these results suggest that PGE(2) induces c-fos and c-jun mRNA expressions through the EP(1) subtype of its receptor. In order to study the functional relevance of these protooncogenes, we then studied the effect of inhibition of their synthesis by the use of antisense oligonucleotide. Alkaline phosphatase (ALP) suppression by 17-phenyl-omega-trinor PGE(2) was reversed by antisense oligonucleotide for either c-fos or c-jun. These results suggest that PGE(2), via the EP(1) subtype of the PGE receptor, negatively modulates the transition from proliferation to the matrix maturation stage through the induction of c-fos and c-jun. However, antisense oligonucleotide for c-fos or c jun did not alter the prostaglandin G/H synthase-2 mRNA expression induced by EP(1). Thus, it is possible that c-fos and c-jun inductions do not account for all the EP(1)-mediated PGE(2) actions in MC3T3-E1 cells. PMID- 10666499 TI - Changes in bone turnover during the menstrual cycle in cynomolgus monkeys. AB - It is well established that estrogen deficiency at menopause results in increased bone turnover, which is reflected in increased concentrations of markers of bone formation and bone resorption in serum and urine. Since serum 17beta-estradiol concentrations vary markedly throughout the menstrual cycle, one would expect to see changes in bone turnover as well. Studies in humans have not yielded consistent results, perhaps because of differences in diet and activity throughout the test period. Therefore, we examined changes in bone biomarkers throughout the menstrual cycle in cynomolgus macaques. Seven intact female cynomolgus macaques (Macaca fascicularis) were evaluated. Vaginal swabs for menstrual blood were performed 3 times/week to determine the stage of the reproductive cycle. Blood and urine were collected at weekly or biweekly intervals for a total of eight samples per monkey for analysis of serum 17beta estradiol, progesterone, parathyroid hormone (PTH), osteocalcin, bone-specific alkaline phosphatase (BSAP), and urinary CrossLapstrade mark. Cycle lengths were determined, and collection days were adjusted to a standardized length of 28 days for all monkeys. Values for bone biomakers were evaluated as % mean for each monkey cycle. By fitting the data to a sine wave (cosinor analysis), serum osteocalcin, BSAP, and urinary CrossLaps demonstrated significant cycles with peaks at days 2.6, 7.3, and 27.8, respectively. Serum osteocalcin and urinary CrossLaps were inversely correlated to serum 17beta-estradiol. Urinary CrossLaps were significantly lower in the week just prior to and during ovulation when estradiol was elevated. No rhythm was detected in serum PTH. A peak in bone resorption when serum 17beta-estradiol is at its nadir is consistent with the hypothesis that estrogen decreases bone turnover. PMID- 10666501 TI - Clinical indications of calcium-phosphate biomaterials and related composites for orthopedic procedures. PMID- 10666500 TI - Histomorphometric evaluation of the recovering effect of human parathyroid hormone (1-34) on bone structure and turnover in streptozotocin-induced diabetic rats. AB - The purpose of this study was to determine whether human parathyroid hormone (h PTH) enhances trabecular bone mass and connectivities that were reduced by streptozotocin (STZ)-induced diabetes in rats. Seven-month-old female Wistar rats were injected with STZ or its vehicle intraperitoneally. All vehicle-injected normal controls were sacrificed 0, 4, 6, 8, 12, 14, and 16 weeks after injection, and one-third of the STZ-injected rats were sacrificed as the baseline controls 4, 6, and 8 weeks after the injection. Eight-week h-PTH (6. 0 microg/kg, 6 times a week) or its vehicle treatment by subcutaneous injection for residual diabetic rats was started 4, 6, or 8 weeks after the STZ injection. The rats' proximal right tibiae were processed for undecalcified Villanueva bone staining sections for bone histomorphometry. Furthermore, changes in trabecular connectivities were determined by node-strut analysis. The decreased cancellous bone volume (BV/TV) and turnover in diabetic rats were recovered in all PTH-treated groups. In node strut analysis, the node-related parameters (N.Nd/TV, NdNd/TV) were significantly increased by PTH when it was administered 4 weeks after STZ injection but were not increased when administration was started after 6 weeks. The results indicated that PTH enhanced bone turnover and bone mass but not trabecular connectivity in the late stage of diabetes in rats. Early treatment of osteoporosis is important in preventing fractures caused by decreased bone strength resulting from decreased trabecular connectivity. PMID- 10666502 TI - The role of calcitriol in the treatment of osteoporosis. PMID- 10666503 TI - The dopamine D(1) receptor agonist SKF-82958 serves as a discriminative stimulus in the rat. AB - We examined the discriminative stimulus effects of the high-efficacy dopamine D(1) receptor agonist (+/-)6-chloro-7, 8-dihydroxy-3-ally1-phenyl-2,3,4,5 tetrahydro-1H-3benzazepine++ + hydrobromide (SKF-82958) in rats trained to discriminate SKF-82958 (0.03 mg/kg) from vehicle in a two-lever food-reinforced drug discrimination task. SKF-82958 produced dose-related increases in responding to the SKF-82958 appropriate lever with full substitution occurring at the training dose. Pretreatment with the dopamine D(1)/D(5) receptor antagonist (-) trans-6,7,7a,8,9, 13b-hexahydro-3-chloro-2hydroxy-N-methyl-5H-benzo-[d]naphtho ?2, 1-b?azepine (SCH-39166) (0.01 mg/kg) attenuated the discriminative stimulus effects of SKF-82958. Pretreatment with the dopamine D(2) receptor antagonist raclopride (0.03 mg/kg) had no effect. The high-efficacy dopamine D(1) receptor agonist R(+)6chloro-7, 8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrobromide (SKF-81297) fully substituted for SKF-82958, whereas the low efficacy dopamine D(1) receptor agonist (+/-)1-phenyl-2,3,4, 5-tetrahydro-(1H)-3 benzazepine-7,8-diol hydrochloride (SKF-38393) produced only partial substitution. The dopamine D(2) receptor agonist trans-(+/-)-4,4a,5,6,7,8,8a, 9 octahydro-5-propyl-1H-propyl-1H-pyrazolo[3,4-g]quinoline dihydrochloride (quinpirole) and the indirect dopamine agonist cocaine did not substitute fully for the SKF-82958 discriminative stimulus cue. These results demonstrate that the high-efficacy dopamine D(1) receptor agonist SKF-82958 can serve as an effective discriminative stimulus in the rat, and that these effects are mediated by a dopamine D(1)-like receptor mechanism. PMID- 10666504 TI - Adrenomedullin decreases extracellular signal-regulated kinase activity through an increase in protein phosphatase-2A activity in mesangial cells. AB - Adrenomedullin is a recently identified peptide hormone that has receptors in a number of different systems including renal mesangial cells. We reported recently that adrenomedullin can cause a decrease in extracellular signal-regulated kinase (ERK) activity and increase jun amino-terminal kinase (JNK) and P38 mitogen activated protein kinase (P38 MAPK) acitivities in rat mesangial cells. Associated with these responses we also reported that adrenomedullin can decrease proliferation and increase apoptosis in mesangial cells. The major aim of the present study was to examine the mechanism of decrease in ERK activity by adrenomedullin and to identify the role of protein phosphatase 2A (PP2A) in the decrease in ERK activity, using okadaic acid [9,10-Deepithio-9,10 didehydroacanthifolicin], a selective inhibitor of PP2A at low nanomolar concentrations. The adrenomedullin-induced decrease in [3H]-thymidine incorporation and increase in apoptosis were reversed by okadaic acid at the concentration that selectively inhibits PP2A. Okadaic acid completely reversed the ERK inhibition caused by adrenomedullin, suggesting that PP2A may be involved in the adrenomedullin-mediated changes in proliferation, apoptosis and ERK activity. PP2A activity in mesangial cells was increased over time following exposure to adrenomedullin. The tyrosine phosphorylation of ERK did not change significantly following adrenomedullin treatment although the ERK activity was decreased significantly. This suggests that the decrease in ERK activity is not mediated through a decrease in MEK (a dual phosphorylating kinase upstream of ERK) or by an increase in MKP-1/2 (a dual specificity phosphatase) activities. Thus we conclude that the mechanism of adrenomedullin-induced decrease in ERK activity in rat mesangial cells is at least in part mediated by an increase in PP2A activity. PMID- 10666505 TI - sigma receptor ligands attenuate N-methyl-D-aspartate cytotoxicity in dopaminergic neurons of mesencephalic slice cultures. AB - We investigated the potential neuroprotective effects of several sigma receptor ligands in organotypic midbrain slice cultures as an excitotoxicity model system. When challenged with 100-microM N-methyl-D-aspartate (NMDA) for 24 h, dopaminergic neurons in midbrain slice cultures degenerated, and this was prevented by (5R, 10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,b]-cyclohepten-5, 10-imine (MK-801; 1-10 microM). Concomitant application of ifenprodil (1-10 microM) or haloperidol (1-10 microM), both of which are high-affinity sigma receptor ligands, significantly attenuated the neurotoxicity of 100 microM NMDA. The sigma(1) receptor-selective ligand (+)-N-allylnormetazocine ((+)-SKF 10047; 1 10 microM) was also effective in attenuating the toxicity of NMDA. The effect of R(-)-N-(3-phenyl-1-propyl)-1-phenyl-2-aminopropane hydrochloride ((-)-PPAP), a sigma receptor ligand with negligible affinity for the phencyclidine site of NMDA receptors, was also examined. (-)-PPAP (3-100 microM) caused a concentration dependent reduction of NMDA cytotoxicity, with significant protection at concentrations of 30 and 100 microM. In contrast, (+)-SKF 10047 (10 microM) and ( )-PPAP (100 microM) showed no protective effects against cell death induced by the Ca(2+) ionophore ionomycin (1-3 microM). These results indicate that sigma receptor ligands attenuate the cytotoxic effects of NMDA on midbrain dopaminergic neurons, possibly via inhibition of NMDA receptor functions. PMID- 10666506 TI - Effect of glipizide on dopamine synthesis, release and metabolism in PC12 cells. AB - Sulfonylureas block ATP-dependent K(+) channels (K/ATP channels) in pancreatic beta cells and brain gamma-aminobutyric acid (GABA) containing neurons causing depolarization-evoked insulin or GABA release. In high concentrations, sulfonylureas also inhibit catecholamine release from bovine adrenal chromaffin cells and isolated guinea pig aorta. In this study, we examined the effect of glipizide, a sulfonylurea, on dopamine release from PC12 cells and found that neither basal nor K(+)-stimulated dopamine release was affected. Although PC12 cells expressed mRNA for the K/ATP channel, functional K/ATP channels could not be demonstrated electrophysiologically, consistent with the lack of effect of glipizide on dopamine release. Glipizide did, however, increase cytoplasmic retention of the acidic dopamine metabolites, 3, 4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), indicating blockade of their outward transport. The cellular accumulation of DOPAC was accompanied by reduced tyrosine hydroxylase activity and reduced formation of dopamine and its metabolites presumably by a negative feedback effect of the increased cytoplasmic concentrations of DOPAC. PMID- 10666507 TI - Characterization and modulation of [125I]iberiotoxin-D19Y/Y36F binding in the guinea-pig urinary bladder. AB - The radioligand binding characteristics of the Ca(2+)-activated K(+) channel ligand [125I]iberiotoxin-D19Y/Y36F were examined in guinea-pig urinary bladder membranes. Saturation analysis revealed a single class of high affinity binding sites in the bladder with a K(D) value of 45.6 pM and a B(max) value of 112 fmol/mg protein. Specific binding was displaced by unlabeled iberiotoxin and penitrem A, but not by blockers of other classes of K(+) channels including alpha dendrotoxin, margatoxin and apamin. The indole alkaloids, paxilline and verruculogen, significantly increased binding by 4.5- and 4.3-fold, respectively. Tetraacetic acid derivatives such as ethylenediamine tetraacetic acid and ethyleneglycoltetraacetic acid enhanced specific [125I]iberiotoxin-D19Y/Y36F binding about 2.5-fold, which was not attributable to calcium chelation. This increase was due to a significant change in ligand binding affinity (K(D)=6.3 pM), but not due to a change in the B(max), indicating that these compounds may enhance toxin binding via allosteric interactions. Collectively, these results demonstrate that the binding sites for [125I]iberiotoxin-D19Y/Y36F present in the urinary bladder shows a pharmacological profile typical of maxi-K(+) channels and can be modulated, not only by previously known indole alkaloids, but also by tetraacetic acid analogs. PMID- 10666508 TI - Topiramate potentiates the antiseizure activity of some anticonvulsants in DBA/2 mice. AB - Topiramate (1-50 mg/kg, intraperitoneally (i.p.)) was able to antagonize audiogenic seizures in DBA/2 mice in a dose-dependent manner. Topiramate at dose of 2.5 mg/kg i.p., which per se did not significantly affect the occurrence of audiogenic seizures in DBA/2 mice, potentiated the anticonvulsant activity of carbamazepine, diazepam, felbamate, lamotrigine, phenytoin, phenobarbital and valproate against sound-induced seizures in DBA/2 mice. The degree of potentiation induced by topiramate was greatest for diazepam, phenobarbital and valproate, less for lamotrigine and phenytoin and not significant for carbamazepine and felbamate. The increase in anticonvulsant activity was associated with a comparable increase in motor impairment. However, the therapeutic index of the combination of all drugs+topiramate was more favourable than that of antiepileptics+ saline, with the exception of carbamazepine or felbamate+topiramate. Since topiramate did not significantly influence the total and free plasma levels of the anticonvulsant drugs studied, we suggest that pharmacokinetic interactions, in terms of total or free plasma levels, are not probable. However, the possibility that topiramate can modify the clearance from the brain of the anticonvulsant drugs studied cannot be excluded. In addition, topiramate did not significantly affect the hypothermic effects of the anticonvulsants tested. In conclusion, topiramate showed an additive effect when administered in combination with some classical anticonvulsants, most notably diazepam, phenobarbital, lamotrigine, phenytoin and valproate. PMID- 10666509 TI - Effects of zatebradine and propranolol on canine ischemia and reperfusion-induced arrhythmias. AB - 1,3,4,5-Tetrahydro-7,8-dimethoxy-3[3-[[2-(3, 4-dimethoxyphenyl) ethyl]methylamino]propyl]-2H-3-benzazepin-2-one -hy drochloride (Zatebradine) is a specific bradycardiac agent, blocking the hyperpolarization-activated pacemaker current (I(f)), and thus has no negative inotropic effect. The purpose of this study was to examine whether zatebradine is effective against ischemia and reperfusion-induced arrhythmias in dogs compared to propranolol. Arrhythmia was induced by ligation of the left anterior descending coronary artery followed by reperfusion. Ischemia-induced biphasic arrhythmias were suppressed in both zatebradine and propranolol groups. During ischemia, fatal ventricular fibrillation occurred in four dogs in the control group, 0 in the zatebradine group, and two dogs in the propranolol group. Of the 31 dogs subjected to reperfusion, mortality rates in the zatebradine, propranolol, and control groups were 56%, 75%, and 86%, respectively, and there were no significant differences. In the heart beating 10 beats/min faster than the predrug heart rate by atrial pacing, both zatebradine and propranolol attenuated ischemia-induced arrhythmias but did not affect reperfusion arrhythmias. Our results suggest that I(f) and/or beta-adrenoceptors rather than the bradycardiac action might be related to the antiarrhythmic effects during ischemia, but that they do not play a role in the generation of the reperfusion-induced ventricular arrhythmias. PMID- 10666510 TI - Characterization of bradykinin B(2) receptor antagonists in human and rat urinary bladder. AB - The effect of three selective bradykinin B(2) receptor antagonists, MEN11270 (H DArg-Arg-Pro-Hyp-Gly-Thi-c(Dab-DTic-Oic-Arg)c(7gamma-1 0alpha)), Icatibant (H DArg-Arg-Pro-Hyp-Gly-Thi-Ser-DTic-Oic-Arg-OH), and FR173567 ((E)-3-(6-acetamido-3 pyridyl)-N-[N-[2, 4-dichloro-3-[(2-methyl-8-quinolinyl) oxymethyl] phenyl]-N methylaminocarbonylmethyl]acrylamide) was evaluated in the human and rat urinary bladder in vitro and in vivo in anaesthetized rats. Bradykinin evoked a concentration-dependent contraction of human (pD(2)=7.2) and rat (pD(2)=7.7) detrusor muscle strips. In human preparations, all the antagonists tested produced a rightward-shift in the concentration-response curve for bradykinin. Schild plot analysis yielded pK(B) values of 8.4, 8.4 and 8.6 for MEN11270, Icatibant, and FR173567, respectively. In the rat preparations the three antagonists (at 100 nM concentration), produced a shift to the right which gave apparent pA(2) values of 8. 2, 8.0 and 8.1 for MEN11270, Icatibant, and FR173567, respectively. In anaesthetized rats, both MEN11270 and Icatibant (1-10 nmol/kg i.v. ) dose dependently reduced the bradykinin (100 nmol/kg i.v.)-induced urinary bladder contraction, their effect being prompt and long-lasting. In contrast, FR173567 (100 nmol/kg i.v.) produced a partial and short-lasting inhibition of bradykinin-induced bladder contractions. The present findings indicate that all the antagonists tested recognize with similar potencies the bradykinin B(2) receptors expressed in the detrusor muscle of both humans and rats. MEN11270 and Icatibant possess a higher potency and longer duration of action in vivo than FR173657, suggesting that the activity of this non-peptide antagonist in vivo is hampered by factors unrelated to its affinity for bradykinin B(2) receptors. PMID- 10666511 TI - Nociceptin inhibits tonic nitric oxide release in the mouse isolated proximal colon. AB - Nociceptin/orphanin FQ, the endogenous ligand of the opioid receptor-like (ORL1) receptor, caused contractions in the isolated colon of the mouse. Tetrodotoxin and the nitric oxide (NO) synthase inhibitor Nomega-nitro-L-arginine also produced contractions which were quantitatively similar to those seen in response to nociceptin. In the presence of either tetrodotoxin or Nomega-nitro-L-arginine, nociceptin was unable to cause a further contraction, whereas the muscarinic receptor agonist carbachol elicited a contractile response. Nociceptin had no contractile activity in colonic preparations contracted by Nomega-nitro-L arginine then relaxed by addition of the NO donor sodium nitroprusside. These data suggest that nociceptin causes contractions of the mouse proximal colon by inhibiting the tonic, neuronal release of NO. PMID- 10666512 TI - The effect of endothelin-1 on lipopolysaccharide-induced cyclooxygenase 2 expression in association with prostaglandin E(2). AB - We demonstrated previously that endothelin-1 (10(-14) to 10(-8) M) promotes lipopolysaccharide-induced cyclooxygenase 2 expression and prostaglandin E(2) production through endothelin ET(B) receptors effects which are up-regulated by lipopolysaccharide. In the present study, we confirmed these findings and showed that prostaglandin E(2) (10(-6) to 10(-5) M) inhibited the lipopolysaccharide plus endothelin-1-induced cyclooxygenase 2 expression more profoundly as compared to its inhibition of the lipopolysaccharide-induced cyclooxygenase 2 expression. The endothelin ET(B) receptor selective antagonist, N-cis-2, 6-dimethylpiperidino carbonyl-L-gamma-methyl-leucyl-D-L-methoxy carbon yl-tryptophanyl-D-norleucine (BQ788), partly inhibited this suppression. Interestingly, the expression of endothelin ET(B) receptors in macrophages was increased by lipopolysaccharide plus prostaglandin E(2) (10(-8) to 10(-5) M) about 1.6-fold compared with that evoked by lipopolysaccharide stimulation alone. We also showed that treatment with endothelin-1 at 10(-14) M (15 min) elevated an intracellular cyclic AMP concentration in macrophages stimulated by lipopolysaccharide or lipopolysaccharide plus prostaglandin E(2) (10(-6) M) for 6 h, and the elevation in the latter cells was more pronounced. These results suggested that endothelin 1 shows an opposite modulation of lipopolysaccharide-induced cyclooxygenase 2 expression in macrophages through endothelin ET(B) receptors, depending on the level of extracellular prostaglandin E(2), and the changes of intracellular cyclic AMP by endothelin-1 may be involved in this mechanism. PMID- 10666513 TI - LY354740 affects startle responding but not sensorimotor gating or discriminative effects of phencyclidine. AB - LY354740 ?(1S,2S,5R,6S)-2-aminobicyclo[3.1.0]hexane-2, 6-dicarboxylate monohydrate?, a selective group II metabotropic glutamate (mGlu) receptor agonist, was recently reported to attenuate the behavioral effects of phencyclidine (PCP) in rats. In the present study, LY354740 failed to attenuate the discriminative stimulus properties of PCP and its disruption of prepulse inhibition of the acoustic startle response, at a dose range which decreased startle responding. The suggestion that mGlu group II receptor activation induces antipsychotic effects may be premature. PMID- 10666514 TI - Tandem sulfur-containing amino acids are epicritical determinants of dopamine D(2) receptor pharmacology. AB - The conserved aspartic acid that is required for ligand binding to the dopamine D(2) receptor is followed by three tandem sulfur-containing amino acids. While previous point mutation studies did not reveal any single one of these residues as being critical for ligand binding, we now show that simultaneously substituting all three with isovolumetric, non sulfur-containing amino acids results in large decreases in the binding affinity for dopamine, (-)-raclopride and 7-(-4(4-(2, 3-dichlorophenyl)-1-piperazinyl)butyloxy)-3, 4-dihydro-2(1H) quinolinone (aripiprazole), but not for methylspiperone or allosteric modulators. PMID- 10666515 TI - Spinal nerve ligation: what to blame for the pain and why. PMID- 10666516 TI - Antagonists of excitatory opioid receptor functions enhance morphine's analgesic potency and attenuate opioid tolerance/dependence liability. AB - Recent preclinical and clinical studies have demonstrated that cotreatments with extremely low doses of opioid receptor antagonists can markedly enhance the efficacy and specificity of morphine and related opioid analgesics. Our correlative studies of the cotreatment of nociceptive types of dorsal-root ganglion neurons in vitro and mice in vivo with morphine plus specific opioid receptor antagonists have shown that antagonism of Gs-coupled excitatory opioid receptor functions by cotreatment with ultra-low doses of clinically available opioid antagonists, e.g. naloxone and naltrexone, markedly enhances morphine's antinociceptive potency and simultaneously attenuates opioid tolerance and dependence. These preclinical studies in vitro and in vivo provide cellular mechanisms that can readily account for the unexpected enhancement of morphine's analgesic potency in recent clinical studies of post-surgical pain patients cotreated with morphine plus low doses of naloxone or nalmefene. The striking consistency of these multidisciplinary studies on nociceptive neurons in culture, behavioral assays on mice and clinical trials on post-surgical pain patients indicates that clinical treatment of pain can, indeed, be significantly improved by administering morphine or other conventional opioid analgesics together with appropriately low doses of an excitatory opioid receptor antagonist. PMID- 10666517 TI - Computer-assisted infrared thermographic study of axon reflex induced by intradermal melittin. AB - The aim of the present study was to investigate whether melittin, the principal toxin of the honeybee (Apis mellifera) venom, can be used as an algogenic agent in the study of pain in humans. Five micrograms of melittin in 0.5 ml of saline was intradermally injected into the volar aspect of the forearm. Resultant pain was scored by a visual analogue scale (VAS), and skin temperature change was analyzed by means of a computer-assisted infrared thermography. Intradermal melittin temporarily produced severe pain, followed by a sustained increase in skin temperature. The skin temperature increase peaked in about 10 min and outlasted 1 h. Topical application of 10% lidocaine gel did not significantly suppress the melittin-induced pain, but markedly suppressed both the increase in the peak temperature and the area of temperature increase. In conclusion, 5 microg of melittin is sufficient to produce pain in humans and 10% lidocaine gel differentially decreases the melittin-induced axon reflex without any significant analgesic effect. PMID- 10666518 TI - Secondary hyperalgesia persists in capsaicin desensitized skin. AB - Several lines of evidence suggest that secondary hyperalgesia to punctate mechanical stimuli arises from central sensitization to the input from primary afferent nociceptors. Conventional C-fiber nociceptors respond to heat stimuli and yet heat hyperalgesia is absent in the region of secondary hyperalgesia. This evidence suggests that the central sensitization to nociceptor input does not involve heat sensitive nociceptors. To test this hypothesis, we investigated whether desensitization of heat sensitive nociceptors by topical application of capsaicin led to an alteration in the secondary hyperalgesia. Two 2x2 cm areas on the volar forearm, separated by 1 cm, were treated in 10 healthy volunteers. One of the areas was desensitized by treatment with 10% topical capsaicin (6 h/day for 2 days). The other site served as vehicle control. Hyperalgesia was produced 2 days later by an intradermal injection of capsaicin (50 microg, 10 microl) at a point midway between the two treatment areas. Secondary hyperalgesia to noxious mechanical stimuli was investigated by using a blade probe (32 and 64 g) attached to a computer-controlled mechanical stimulator. In the area of topical capsaicin treatment, there was a marked increase in heat pain threshold and decrease in heat pain ratings indicating a pronounced desensitization of heat sensitive nociceptors. However, touch threshold and pain to pinching stimuli were not significantly altered. The intradermal capsaicin injection led to the development of a similar degree of secondary hyperalgesia at both the vehicle and capsaicin treatment areas. These results indicate that capsaicin insensitive nociceptive afferents play a dominant role not only in normal mechanical pain but also in secondary hyperalgesia to noxious mechanical stimuli. PMID- 10666519 TI - Effects of intrathecal administration of ziconotide, a selective neuronal N-type calcium channel blocker, on mechanical allodynia and heat hyperalgesia in a rat model of postoperative pain. AB - Ziconotide (SNX-111), a selective blocker of neuronal N-type voltage-sensitive calcium channels, is antinociceptive when it is administered intrathecally. It is currently under clinical investigation for the treatment of malignant and non malignant pain syndromes. The present study was undertaken to compare and contrast antinociceptive properties of ziconotide, morphine and clonidine in a rat model of post-operative pain. Post-operative pain was produced by making a longitudinal incision through the skin, fascia, and muscle of the plantar aspect of the left hindpaw. This procedure produced immediate (0.5 h after surgery) and long-lasting (4-7 days post-surgery) heat hyperalgesia and mechanical allodynia in the injured hindpaw. Pain thresholds in the contralateral hindpaw were unaffected. Administered one day after incisional surgery, intrathecal ziconotide blocked established heat hyperalgesia in the injured hindpaw in a dose-dependent manner yielding an ED(50)4 h) but reversible (<24 h) blockade of established mechanical allodynia. Administered one day after surgery, intrathecal bolus injection of morphine dose-dependently blocked heat hyperalgesia in the injured hindpaw with an ED(50) of 1.6 microg (2.1 nmol) and heat nociceptive responses in the normal hindpaw with an ED(50) of 2.7 microg (3.6 nmol). The effects were immediate and short-lasting ( 10/h or reduced heart rate variability < 70 ms or a positive signal-averaged electrocardiogram) followed by a further arrhythmic risk stratification, obtained through electrophysiologic study. Several published and ongoing trials that utilize various arrhythmic risk stratification techniques as part of their protocol are reviewed. PMID- 10666656 TI - Inpatient versus outpatient antiarrhythmic drug initiation: safety and cost effectiveness issues. AB - Debate exists as to the proper site for initiating antiarrhythmic therapy for supraventricular tachyarrhythmias and other benign forms of ectopy: inpatient versus outpatient. Rapid detection of efficacy and adverse effects, with immediate correction of the latter, favors the inpatient site. Convenience and, under most circumstances, lower cost favor the outpatient site. Circumstances under which adverse event rates, including proarrhythmia, are expectedly low, would favor outpatient initiation. So would the use of an agent whose elimination half-life is so long as to render in-hospital monitoring to steady state highly impractical. Accordingly, outpatient initiation would be suitable for patients without structural heart disease receiving class IC drugs, patients with low risk for torsades de pointes receiving selected class III agents, in whom data in the literature are supportive (as has occurred with sotalol and azimilide), and patients who are to receive amiodarone. Transtelephonic electrocardiographic monitoring can be used to facilitate assessment in the outpatient setting. Inpatient initiation should be considered for patients with underlying sinus node or atrioventricular conduction disturbances, for patients with significant structural heart disease, for patients receiving a drug whose proarrhythmia may be idiosyncratic (e.g., quinidine), and for patients who are to begin an antiarrhythmic drug while in a supraventricular tachyarrhythmia in whom sinus rhythm has not previously been seen. The relative costs and benefits of the approach chosen will be a reflection of the probability that a drug with a chosen mechanism will cause an adverse outcome in a patient with a specific clinical substrate during the period chosen for monitoring. PMID- 10666657 TI - Molecular biology of arrhythmic syndromes. AB - In this review, the up-to-date understanding of the molecular basis of primary ventricular arrhythmias will be outlined. Two disorders have recently been well described at the molecular level, the long QT syndromes and Brugada syndrome, and in this paper we review the current scientific knowledge of each disease. PMID- 10666658 TI - Cost-effective strategies in the acute management of atrial fibrillation. AB - Atrial fibrillation is the most common sustained arrhythmia likely to be encountered in clinical practice. It is associated with significant morbidity and mortality. The treatment of patients with atrial fibrillation can be complex and costly, especially when patients are hospitalized for acute management of this arrhythmia. In this review, we summarize current approaches to the acute management of atrial fibrillation with an emphasis on the most cost-effective approaches. We review acute methods of heart rate control and cardioversion, including pharmacologic and other minimally invasive strategies. We believe that the most cost-effective approaches may require the use of standardized clinical pathways. This may help to ensure that patients with acute atrial fibrillation receive the most effective and efficient care. PMID- 10666659 TI - Catheter ablation in the year 2000. AB - After its introduction in 1987, radiofrequency catheter ablation became established as a safe and effective therapy for the cure of many cardiac arrhythmias in people. The possibility of assessing the relationship between the anatomical target and the electrophysiologic changes produced by radiofrequency pulse delivery has also provided significant improvement in the physician's knowledge of the pathophysiology of the underlying rhythm disturbance. Nowadays, using this therapy, success rates well above 90% with recurrence rates lower than 5% are expected after treatment of most regular supraventricular arrhythmias. As catheter ablation techniques develop, success rates in the range of those obtained for regular supraventricular arrhythmias are expected in the future in the treatment of regular ventricular and irregular supraventricular arrhythmias. PMID- 10666660 TI - New advances in class III antiarrhythmic drug therapy. AB - In the past 2 years, significant advances have been made in class III antiarrhythmic drug therapy. In patients with ventricular arrhythmias and implantable cardioverter defibrillators (ICDs), antiarrhythmic agents are increasingly being used as adjunct therapy to decrease the frequency of ICD discharges. Sotalol was recently shown to be effective in reducing tachyarrhythmias in patients with ICDs. Intravenous amiodarone is being used for the acute treatment of unstable ventricular arrhythmia and is being investigated for the treatment of acute out-of-hospital cardiac arrest. Class III agents are increasingly being used for prophylaxis in patients who have atrial fibrillation or atrial flutter, and data point to an important role for these agents in reducing supraventricular tachyarrhythmias after cardiac surgery. Future studies will need to directly compare these agents with pure anti-adrenergic maneuvers in postoperative patients. In addition to terminating atrial fibrillation and atrial flutter, ibutilide significantly reduces human atrial defibrillation thresholds and increases the percentage of patients who can be cardioverted from atrial fibrillation to sinus rhythm. The US Food and Drug Administration is expected to approve dofetilide for clinical use soon, and it is currently reviewing azimilide (which seems to be devoid of frequency-dependent effects on repolarization) for prophylaxis against atrial fibrillation and atrial flutter. Dronedarone, tedisamal, and trecetilide are now under active study intended to determine their usefulness in patients with cardiac arrhythmias. Experimental studies are ongoing to identify pharmacologic agents that will selectively prolong repolarization in the atria without exerting electrophysiologic effects in the ventricles. PMID- 10666661 TI - Classification system of atrial fibrillation. AB - A number of publications and clinical trials on the management of atrial fibrillation (AF) deal with this arrhythmia as if it represents a single entity. As a result, advances made in recent years have not affected the way AF patients are treated in general practice except, perhaps, for the use of warfarin in anticoagulation. Therefore, there is a need for a classification system and for uniformity in the nomenclature used. The two terms currently used to describe AF, paroxysmal and chronic, require a time frame. It is proposed that if an AF episode lasts longer than 7 days, the condition should be considered chronic. For the first symptomatic, non-self-terminating episode that is fewer than 7 days long, the term recent onset AF may be used, or recent discovery if the AF is asymptomatic or if the duration cannot be determined. Attacks of paroxysmal AF may differ in their duration, frequency, and functional tolerance. In the classification system described, three clinical aspects of paroxysmal AF were isolated in such a way as to have implications for therapy. This classification system was found to be useful for characterizing different subsets of patients with AF. PMID- 10666662 TI - Clinical implication of antiembolic trials in atrial fibrillation and role of transesophageal echocardiography in atrial fibrillation. AB - Risk for stroke in patients with atrial fibrillation (AF) is highly heterogeneous. Increasing age, history of diabetes, hypertension, previous transient ischemic attack or stroke, and poor ventricular function are independent risk factors for stroke in patients with AF. Accordingly, some groups of patients with AF have low risk and some have high risk. In general, patients at high risk benefit most from anticoagulation therapy with warfarin. In general, if a patient is younger than 65 years of age and has none of the defined risk factors, the stroke rate without prophylaxis (aspirin or warfarin) is low. In patients 65 to 75 years of age with no risk factors, the risk for stroke is low with either aspirin or warfarin therapy; the choice is left to the caretaking physician. All patients older than 75 years and all patients of any age who have risk factors obtain striking benefit from the use of anticoagulation with warfarin. This benefit far outweighs any risk for major hemorrhage. PMID- 10666663 TI - Amiodarone: clinical trials. AB - Amiodarone is an antiarrhythmic agent commonly used in the treatment of supraventricular and ventricular tachyarrhythmias. This article reviews the results and clinical implications of primary and secondary prevention trials in which amiodarone was used in one of the treatment arms. Key post-myocardial infarction primary prevention trials include the European Myocardial Infarct Amiodarone Trial (EMIAT) and the Canadian Amiodarone Myocardial Infarction Trial (CAMIAT), both of which demonstrated that amiodarone reduced arrhythmic but not overall mortality. In congestive heart failure patients, amiodarone was studied as a primary prevention strategy in two pivotal trials: Grupo de Estudio de la Sobrevida en la Insuficiencia Cardiac en Argentina (GESICA) and Amiodarone in Patients With Congestive Heart Failure and Asymptomatic Ventricular Arrhythmia (CHF-STAT). Amiodarone was associated with a neutral overall survival and a trend toward improved survival in nonischemic cardiomyopathy patients in CHF/STAT and improved survival in GESICA. In post-myocardial infarction patients with nonsustained ventricular tachycardia and a depressed ejection fraction, the Multicenter Automatic Defibrillator Implantation Trial (MADIT) demonstrated that implantable cardioverter-defibrillators (ICD) statistically improved survival compared to the antiarrhythmic drug arm, most of whose patients were taking amiodarone. In patients with histories of sustained ventricular tachycardia or ventricular fibrillation, the Cardiac Arrest Study in Seattle: Conventional Versus Amiodarone Drug Evaluation (CASCADE) trial demonstrated that empiric amiodarone lowered arrhythmic recurrence rates compared to other drugs guided by serial Holter or electrophysiologic studies. However, arrhythmic death rates were high in both treatment arms of the study. Several secondary prevention trials, including the Antiarrhythmics Versus Implantable Defibrillators Study (AVID), the Canadian Implantable Defibrillator Study (CIDS), and the Cardiac Arrest Study Hamburg (CASH), have demonstrated the superiority of ICD therapy compared to empiric amiodarone in improving overall survival. Based on the above findings, amiodarone is safe to use in post-myocardial infarction and congestive heart failure patients that need antiarrhythmic therapy. Although amiodarone is effective in treating malignant arrhythmias, high-risk patients should be considered for an ICD as frontline therapy. PMID- 10666664 TI - Bibliography. Current world literature. Arrhythmias. PMID- 10666665 TI - Molecular abnormalities of the brain in Down syndrome: relevance to Alzheimer's neurodegeneration. AB - Down syndrome is caused by over-expression of genes located within a segment of chromosome 21, termed the Down locus. Down syndrome is associated with developmental abnormalities of the central nervous system that result in mental retardation and age-dependent Alzheimer-type neurodegeneration. Some of the neurodegenerative lesions, including A beta amyloid deposition, apoptotic cell death, and aberrant dendritic arborization, are in part due to constitutively increased expression of genes that encode the amyloid precursor protein, superoxide dismutase I, and S100-beta, and located within the Down locus. However, neurodegeneration in Down syndrome is also associated with aberrant expression of genes that are not linked to the Down locus, including the growth associated protein, GAP-43, nitric oxide synthase 3, neuronal thread protein, and pro-apoptosis genes such as p53, Bax, and interleukin-1 beta-converting enzyme. Increased expression of these non-Down locus genes correlates with proliferation of dystrophic neurites and apoptotic cell death, two important correlates of cognitive impairment in Alzheimer's disease. This article reviews the functional importance of abnormal gene expression in relation to Alzheimer-type neurodegeneration in brains of individuals with Down syndrome. PMID- 10666667 TI - The Down syndrome critical region. AB - Since the early 1970's numerous attempts have been made to learn whether specific segments of chromosome 21, when triplicated, are responsible for the clinical condition Down syndrome (DS). Studies were reported in which positive or negative clinical diagnoses of DS were made in the presence of partial trisomy of one or another segment of the chromosome. The distal half of the long arm of 21 (21q22) possesses most of the gene transcribing sites of the chromosome. It was this region that was thought to contain loci essential to production of the clinical syndrome. Subsequent studies identified subregions of this band as "minimal" or "critical" sites necessary and sufficient to produce the clinical condition. A major problem with these assignments was that different investigators defined different critical/minimal regions. In 1994 evidence was presented in which regions of most of the long arm of chromosome 21 were said to contribute to the DS phenotype. Soon after, a report described a child with DS and partial tetrasomy of the short arm and proximal long arm of 21, segments clearly distinct from the previously identified critical areas. Thus the clinical diagnosis of DS can be made in the presence of partial aneuploidy of nearly all segments of chromosome 21. It must be concluded that no evidence exists that individual loci on 21 are singularly responsible for specific phenotypic abnormalities in DS. Without exception, each of the clinical findings associated with DS is a multifactorial trait. The analysis of each trait in DS should thus be similar to analyses of the same traits in the general population with a focus on the way aneuploidy affects expression of multifactorial characteristics. PMID- 10666666 TI - Reduced aldehyde dehydrogenase levels in the brain of patients with Down syndrome. AB - Aldehyde dehydrogenase (ALDH) is a key enzyme in fructose, acetaldehyde and oxalate metabolism and represents a major detoxification system for reactive carbonyls and aldehydes. In the brain, ALDH exerts a major function in the metabolism of biogenic aldehydes, norepinephrine, dopamine and diamines and gamma aminobutyric acid. Subtractive hybridization studies in Down Syndrome (DS) fetal brain showed that mRNA for ALDH are downregulated. Here we studied the protein levels in the brain of adult patients. The proteins from five brain regions of 9 aged patients with DS and 9 controls were analyzed by two-dimensional (2-D) gel electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry. ALDH levels were reduced in the brain regions of at least half of the patients with Down Syndrome, as compared to controls. The decreased ALDH levels in the DS brain may result in accumulation of aldehydes which can lead to the formation of plaques and tangles reflecting abnormally cross-linked, insoluble and modified proteins, found in aged DS brain. Furthermore, we constructed a 2-Dmap including approximately 120 identified human brain proteins. PMID- 10666668 TI - Neuropathology. PMID- 10666669 TI - c-fos expression in brains of patients with Down syndrome. AB - c-fos is a protooncogene serving in multiple physiological processes in brain from signalling to proliferation and synaptic plasticity. We therefore decided to determine this transcription factor in control and Down Syndrome (DS) brain with the Rationale that c-fos may be linked to brain damage in DS. We determined mRNA steady state levels in frontal, parietal, occipital, temporal cortex and cerebellum of 9 patients with DS and 9 controls using RT-PCT. Significantly increased levels of mRNA c-fos normalized versus the housekeeping gene beta-actin mRNA were found in frontal, parietal and temporal cortex of DS brain. c-fox mRNA levels comparable to controls were found in occipital cortex and cerebellum. Deteriorated c-fos expression in the individual brain regions may be linked to increased apoptosis and neurodegeneration, overexcitation by excitatory amino acids of reactive oxygen species. PMID- 10666670 TI - Neuronal cell death in Down's syndrome. AB - Down's syndrome (DS), occurring in 0.8 out of 1,000 live births, is a genetic disorder in which an extra portion of chromosome 21 leads to several abnormalities. With respect to the nervous system, it causes mental retardation. It is conceived that abnormal neuronal cell death in development is involved, but there is no direct evidence yet. In addition to developmental brain abnormalities, almost all DS brains over 40 years old manifest a similar pathology to Alzheimer's disease (AD), including the presence of senile plaques (SP) and neurofibrillary tangles (NFT). Although there was a debate to segregate dementia from underlying mental retardation, at least some portion of DS patients exhibit deteriorated mental status with aging. The mechanism underlying these abnormalities at the molecular level remains to be elucidated. Recently there have been several reports suggesting abnormalities reflecting increased risk to apoptosis in DS brains. Increased expression of several apoptosis-related genes (p53, fas, ratio of bax to bcl-2, GAPDH) in DS brains has been reported. Cultured neurons from both patients and model animals are reportedly more vulnerable to apoptosis. Overproduction of reactive oxygen species and its causative roles for increased apoptosis in DS tissues are suggested. One possible hypothesis is an increased susceptibility to apoptosis due to p53 overactivation in DS brains. A beta 42, a critical peptide for AD pathology from amyloid precursor protein (APP), can be detected in DS brains. A beta 42 is deposited in SP from an early stage, suggesting common molecular mechanisms in DS and AD. Animal models for DS are important in the search of molecular mechanisms. Several types of models are now available. Future DS studies are expected to integrate information from animal models and human tissues. PMID- 10666671 TI - Altered gene expression in fetal Down syndrome brain as revealed by the gene hunting technique of subtractive hybridization. AB - Information on gene expression in brain of patients with Down Syndrome (DS, trisomy 21) is limited and molecular biological research is focussing on mapping and sequencing chromosome 21. The information on gene expression in DS available follows the current concept of a gene dosage effect due to a third copy of chromosome 21 claiming overexpression of genes encoded on this chromosome. Based upon the availability of fetal brain and recent technology of gene hunting, we decided to use subtractive hybridization to evaluate differences in gene expression between DS and control brains. Subtractive hybridization was applied on two fetal brains with DS and two age and sex matched controls, 23rd week of gestation, and mRNA steady state levels were evaluated generating a subtractive library. Subtracted sequences were identified by gene bank and assigned by alignments to individual genes. We found a series of up- and downregulated sequences consisting of chromosomal transcripts, enzymes of intermediary metabolism, hormones, transporters/channels and transcription factors (TFs). We show that trisomy 21 or aneuploidy leads to the deterioration of gene expression and the derangement of transcripts describes the impairment of transport, carriers, channels, signaling, known metabolic and hormone imbalances. The dys coordinated expression of transcription factors including homeobox genes, POU domain TFs, helix-loop-helix-motifs, LIM domain containing TFs, leucine zippers, forkhead genes, maybe of pathophysiological significance for abnormal brain development and wiring found in patients with DS. This is the first description of the concomitant expression of a large series of sequences indicating disruption of the concerted action of genes in this disorder. PMID- 10666672 TI - Gene expression in fetal Down syndrome brain as revealed by subtractive hybridization. AB - Information on gene expression in brain of patients with Down Syndrome (DS, trisomy 21) is limited and molecular biological research is focussing on mapping and sequencing chromosome 21. The information on gene expression in DS available follows the current concept of a gene dosage effect due to a third copy of chromosome 21 claiming overexpression of genes encoded on this chromosome. Based upon the availability of fetal brain and recent technology of gene hunting, we decided to use subtractive hybridization to evaluate differences in gene expression between DS and control brains. Subtractive hybridization was applied on two fetal brains with DS and two age and sex matched controls, 23rd week of gestation, and mRNA steady state levels were evaluated generating a subtractive library. Subtracted sequences were identified by gene bank and assigned by alignments to individual genes. We found a series of up- and downregulated sequences consisting of chromosomal transcripts, enzymes of intermediary metabolism, hormones, transporters/channels and transcription factors (TFs). We show that trisomy 21 or aneuploidy leads to the deterioration of gene expression and the derangement of transcripts described describes the involvement of chromosomes other than chromosome 21, explains impairment of transport, carriers, channels, signaling, known metabolic and hormones imbalances. The dys-coordinated expression of transcription factors including homeobox genes, POU-domain TFs, helix-loop-helix-motifs, LIM domain containing TFs, leucine zippers, forkhead genes, maybe of pathophysiological significance for abnormal brain development and wiring found in patients with DS. This is the first description of the concomitant expression of a large series of sequences indicating disruption of the concerted action of genes in that disorder. PMID- 10666673 TI - Molecular misreading of genes in Down syndrome as a model for the Alzheimer type of neurodegeneration. AB - The occurrence of +1 frameshifted proteins, such as amyloid precursor protein (APP+1) and ubiquitin-B (UBB+1) in Down syndrome (DS) has been linked to the onset of Alzheimer's disease (AD). In DS and AD patients, but also in elderly non demented persons, these co-called +1 proteins accumulate in the neuropathological hallmarks (neurofibrillary tangles, dystrophic neurites of the neuritic plaques and neuropil threads) and may have deleterious effects on neuronal function. Frameshifts are caused by dinucleotide deletions in GAGAG motifs in messenger RNA and are now thought to be the result of unfaithful transcription of normal DNA by a novel process termed "molecular misreading". In the present review some of the critical events in molecular misreading are discussed, the emphasis being on DS. PMID- 10666674 TI - Expression of the dihydropyrimidinase related protein 2 (DRP-2) in Down syndrome and Alzheimer's disease brain is downregulated at the mRNA and dysregulated at the protein level. AB - Deteriorated migration, axonal pathfinding and wiring of the brain is a main neuropathological feature of Down Syndrome (DS). Information on the underlying mechanisms is still limited, although basic functions of a series of growth factors, cell adhesion molecules, guidance factors and chemoattractants for brain histogenesis have been reported. We used proteomics to detect differences in protein expression between control, DS and Alzheimer's disease brains: In five individual brain regions of 9 individuals of each group we performed two dimensional electrophoresis with MALDI--identification of proteins and determined mRNA levels of DRP-2. Significantly decreased mRNA levels of DRP-2 in four brain regions of patients with DS but not with AD as compared to controls were detected. 2D electrophoresis revealed variable expression of DRP-2 proteins, which showed a high heterogeneity per se. Dysregulation of DRP-2 was found in brains of patients with DS and AD presenting with an inconsistent pattern, which in turn may reflect the inconsistent neuropathological findings in patients with DS and AD. The decrease of mRNA DRP-2 steady state levels in DS along with deteriorated protein expression of this repulsive guidance molecule of the semaphorin/collapsin family, may help to explain deranged migration and histogenesis of DS brain and wiring of AD brain. PMID- 10666675 TI - Gene expression relevant to Down syndrome: problems and approaches. AB - The long arm of human chromosome 21 likely contains several hundred genes. To determine which of these are responsible for specific aspects of the Down Syndrome phenotype, protein functional analysis coupled to phenotypic analysis of transgenic mice will be required. Because such experiments are both time consuming and expensive, prioritizing 21q genes for further studies would be advantageous. Here, we discuss expression analysis, specifically the use of Northern analysis, cDNA array screening and RNA tissue in situ hybridization to assess place and time of expression of forty-two genes. For a subset of these, over expression in normal versus trisomy cell lines and mouse tissues is discussed. Lastly, several examples of alternative processing and their potential for generation of brain specific proteins are described. Together, these experiments give information on time, place and level of expression of a number of 21q genes and suggest some interesting candidates worth further investigation for relevance to Down Syndrome. These data also illustrate the complexities and ambiguities inherent in interpretation and use of expression information. PMID- 10666676 TI - Isolation and analysis of chromosome 21 genes potentially involved in Down syndrome. PMID- 10666677 TI - Differential display reveals deteriorated mRNA levels of NADH3 (complex I) in cerebellum of patients with Down syndrome. AB - Although gene hunting has been carried out in Down Syndrome (DS) cells, information on expressional differences in DS brain is limited. We have recently described expressional differences in fetal DS brain but cannot assign these findings to "DS per" se or simply to "neurodegeneration". We therefore performed gene hunting in cerebellum of adult patients with DS and Alzheimer's disease (AD) neuropathology, AD and controls. The gene hunting method used was differential display and pools of the individual groups were examined to rule out allelic differences. Differential display revealed the absence of a band, identified by sequencing and gene bank work as matching the NADH3 gene (99.1% identity) in cerebellum of DS patients. Dot blots showed the presence of NADH3 signals in only two out of 7 DS patients. We show at the transcriptional level that a mitochondrial enzyme, the complex I, NADH3, is significantly downregulated in DS cerebellum. This extends previous work on deficiencies of the electron transport chain in platelets of patients with DS. PMID- 10666678 TI - Serotonin (5-HT) in brains of adult patients with Down syndrome. AB - Down syndrome (DS) is a genetic disease with developmental brain abnormalities resulting in early mental retardation and precocious, age dependent Alzheimer type neurodegeneration. Furthermore, non-cognitive symptoms may be a cardinal feature of functional decline in adults with DS. As the serotonergic system plays a well known role in integrating emotion, cognition and motor function, serotonin (5-HT) and its main metabolite, 5-hydroxyindol-3-acetic acid (5-HIAA) were investigated in post-mortem tissue samples from temporal cortex, thalamus, caudate nucleus, occipital cortex and cerebellum of adult patients with DS, Alzheimer's disease (AD) and controls by use of high performance liquid chromatography (HPLC). In DS, 5-HT was found to be age-dependent significantly decreased in caudate nucleus by 60% (DS: mean +/- SD 58.6 +/- 28.2 vs. Co: 151.7 +/- 58.4 pmol/g wet tissue weight) and in temporal cortex by about 40% (196.8 +/- 108.5 vs. 352.5 +/- 183.0 pmol/g), insignificantly reduced in the thalamus, comparable to controls in cerebellum, whereas occipital cortex showed increased levels (204.5 +/- 138.0 vs. 82.1 +/- 39.1 pmol/g). In all regions of DS samples, alterations of 5-HT were paralleled by levels of 5-HIAA, reaching significance compared to controls in thalamus and caudate nucleus. In AD, 5-HT was insignificantly reduced in temporal cortex and thalamus, unchanged in cerebellum, but significantly elevated in caudate nucleus (414.3 +/- 273.7 vs. 151.7 +/- 58.4 pmol/g) and occipital cortex (146.5 +/- 76.1 vs. 82.1 +/- 39.1 pmol/g). The results of this study confirm and extend putatively specific 5-HT dysfunction in basal ganglia (caudate nucleus) of adult DS, which is not present in AD. These findings may be relevant to the pathogenesis and treatment of cognitive and non cognitive (behavioral) features in DS. PMID- 10666679 TI - Mechanisms of neuronal death in Down's syndrome. AB - There is growing evidence that neuronal death in Down's syndrome is due to apoptotic mechanisms. The phenomena, however, that trigger and regulate programmed cell death in the Down's syndrome-related neurodegeneration are still much debated. In vitro evidence has suggested that the main factor responsible for neuronal death in this condition is the accumulation of beta-amyloid, due to the overexpression of its precursor protein. Another hypothesis argues for the importance of reactive oxygen species in neuronal death. However, the in vivo findings do not entirely support either theories. We propose that neuronal apoptosis, as well as the formation of Alzheimer-type pathology, in Down's syndrome is due to an aberrant re-entry of neurones into the cell division cycle. Due to the simultaneous overexpression of conflicting cell cycle regulatory signals the mitogenic amyloid precursor and the differentiation factor S100, the cell cycle is abandoned. Subsequently the cell cycle arrest may lead to either the formation of Alzheimer-related pathology or to apoptotic cell death. PMID- 10666680 TI - Impaired brain glucose metabolism in patients with Down syndrome. AB - A series of impaired metabolic functions in Down Syndrome (DS) including glucose handling has been described. Recent information from positron emission tomography studies in DS patients and our finding of downregulated phosphoglucose isomerase (PGI) in fetal brain with DS by gene hunting using subtractive hybridization, made us investigate PGI, a key enzyme of glucose metabolism, in brain of patients with DS, Alzheimer's disease (AD) and controls. PGI and phosphofructokinase (PFK) activities were determined in frontal, parietal, temporal, occipital lobe and cerebellum of 9 controls, 9 patients with DS and 9 patients with AD. PGI activity in DS brain was significantly decreased in frontal, temporal lobe and cerebellum, comparable to controls in parietal lobe and elevated in occipital lobe. Brain PGI activity of patients with AD was comparable to controls in all regions tested, PFK, a rate limiting enzyme of glucose metabolism, was comparable between all brain regions of all three groups. Data of this study confirm impaired glucose metabolism in DS proposed in literature and found by positron emission tomography (PET) studies. We show that changes in glucose handling in patients with AD as evaluated by PET studies are not supported by our data, although not contradictory, as determinants other than glucose metabolizing enzymes as e.g. vascular factors and glucose transport may account for these findings. Changes of downregulated PGI found by subtractive hybridization at the transcriptional level in fetal DS brain along with our findings in DS brain regions suggest a strong specific link between glucose metabolism and DS rather than AD. PMID- 10666681 TI - Oxidative stress and neural dysfunction in Down syndrome. AB - Total or partial trisomy of chromosome 21 occurs with relatively high frequency and is responsible for the occurrence of Down syndrome. Phenotypically, individuals with Down syndrome display characteristic morphological features and a variety of clinical disorders. One of the challenges for researchers in this field has been to ascertain and understand the relationship between the Down syndrome phenotype with the gene dosage effect resulting from trisomy of chromosome 21. Much attention therefore, has been given towards investigating the consequences of overexpressing chromosome 21-linked genes. In particular, an extensive analysis of SOD1 and APP have provided important insights as to how perturbations in the expression of their respective genes may contribute to the Down syndrome phenotype. In this review we will highlight studies which support a key role for SOD1 and APP in the pathogenesis of neural abnormalities observed in individuals with Down syndrome. Central to this relationship is how the redox state of the cell is affected and its consequences to neural function and integrity. PMID- 10666682 TI - Expression of the transcription factor ETS2 in brain of patients with Down syndrome--evidence against the overexpression-gene dosage hypothesis. AB - Overexpression of the transcription factor ETS2 and other genes localized in the socalled critical Down Syndrome region of chromosome 21 due to a gene dosage effect, is an attractive hypothesis for the explanation of the Down Syndrome phenotype. The overexpression of ETS2, however, has never been demonstrated in a human organ. We therefore challenged this hypothesis determining ETS2 levels in several brain regions of patients with Down Syndrome as compared to controls. We used a highly sensitive and quantitative RT-PCR method for the determination of ETS2 mRNA steady state levels in frontal, parietal, temporal, occipital lobe and cerebellum of 9 adult Down Syndrome patients and 9 adult controls. Significantly decreased ETS2 mRNA steady state levels (16.9 +/- 26.7 attogram mRNA ETS2/10 ng total RNA versus 87.7 +/- 92.9 in controls) in frontal lobe of Down Syndrome brain and decreased ETS2 mRNA steady state levels (6.99 +/- 6.4 attogram mRNA ETS2/100 pg beta-actin versus 19.8 +/- 15.7 in controls) in temporal lobe of Down Syndrome brain were found. In the other brain regions no statistically significant difference was detected. Our data provide evidence against the overexpression hypothesis for the development of the Down Syndrome phenotype. Decreased ETS2 transcripts found in temporal and frontal lobe of patients with Down Syndrome, however, may be involved in the pathogenesis of Down Syndrome including specific neurodegenerative processes and deteriorated plasticity of the brain taking place in Down Syndrome brain, as the concerted action of transcription factors may be seriously impaired. PMID- 10666683 TI - Neuronal apoptosis inhibitory protein (NAIP)-like immunoreactivity in brains of adult patients with Down syndrome. AB - In Down syndrome (DS), enhanced apoptosis (programmed cell death) may play a role in the pathogenesis of characteristic early mental retardation and precocious neurodegeneration of Alzheimer-type. The human IAP (inhibitor of apoptosis proteins) genes (NAIP, c-IAP-2/HIAP-1, c-IAP-1/Hiap-2, XIAP, survivin) are an evolutionary conserved family of proteins which prevent cell death across species, implying that they act at a central, highly conserved point in the cell death cascade. Evidence for downregulation of NAIP-mRNA in fetal DS (23rd week of gestation), as found by subtractive hybridization technique challenged studies at the protein level in adult DS brain specimen. NAIP-like immunoreactivity was determined in four different regions of cerebral cortex and cerebellum in 9 adult DS patients with Alzheimer-like neuropathologic lesions, 9 Alzheimer disease (AD) patients as compared to 9 controls. For the first time, NAIP-IR could be demonstrated in different cortical regions of the human brain. Compared to control subjects, western blotting demonstrated significantly decreased levels in parietal and occipital cortex in DS and in frontal and occipital cortex in AD. While the mode of NAIP action is unknown, inhibition of certain caspases has already been demonstrated for other IAP-family members (c-IAP1, c-IAP2 and XIAP). Although decreased NAIP-IR of certain brain regions in DS and AD awaits further confirmation, the results suggest that alterations of apoptosis regulatory (inhibitory) proteins may be another feature of neurodegeneration in DS and AD. PMID- 10666684 TI - The "gene dosage effect" hypothesis versus the "amplified developmental instability" hypothesis in Down syndrome. AB - Two hypotheses exist to explain the Down syndrome (DS) phenotype. The "gene dosage effect" hypothesis states that the phenotype is a direct result of the cumulative effect of the imbalance of the individual genes located on the triplicated chromosome or chromosome region. In a nut shell, the phenotype results directly from the overexpression of specific chromosome 21 genes. The "amplified developmental instability" hypothesis contends that most manifestations of DS may be interpreted as the results of a non-specific disturbance of chromosome balance, resulting in a disruption of homeostasis. This hypothesis was proposed in an attempt to explain the similarities between the phenotypes of different aneuploid states and the observation that all of the phenotypic traits in DS are also seen in the general population but at lower frequency, with less severity and usually only present as a single trait. Herein, we review recent data and present evidence to support the theory that the phenotypic traits of aneuploid syndromes, and DS in particular, result from the increased dosage of genes encoded on the triplicated chromosome. PMID- 10666685 TI - Downregulation of the transcription factor scleraxis in brain of patients with Down syndrome. AB - Performing gene hunting in fetal Down Syndrome (DS) brain, we found a downregulated sequence with 100% homology to the basic-helix-loop-helix transcription factor (TF) scleraxis (Scl). It was the aim of the study to evaluate Scl-mRNA steady state levels in adult DS brain with Alzheimer's disease (AD) neuropathological changes, brain of patients with AD, and controls in order to find out whether Scl-downregulation is linked to DS per se or simply to neurodegeneration, common to both disorders. Determination of Scl-mRNA steady state levels was carried out by a blotting method in frontal, parietal, temporal, occipital lobe and cerebellum. We found significantly decreased Scl-transcripts in brain of DS and AD, both, when normalized versus the house-keeping gene beta actin or total RNA. We demonstrate the significant decrease of Scl-mRNA steady state levels in the pathogenesis of DS and AD suggesting a tentative role for this transcription factor in the development of the neurodegenerative processes known to occur in both disorders. More specifically, the biological meaning of the downregulation of Scl may be the involvement in the pathogenesis of impaired neuronal plasticity and wiring observed in DS and AD, phenomena regulated by the concerted action of the many transcription factors expressed in human brain. PMID- 10666686 TI - Heat-shock protein 70 levels in brain of patients with Down syndrome and Alzheimer's disease. AB - Heat-shock proteins are proteins serving as molecular chaperones, involved in the protection of cells from various forms of stress. Since the expression of these proteins is closely related to that of amyloid precursor protein (APP), heat shock protein has been studied in brain of patients with Alzheimer's disease (AD) and furthermore, brain Hsp70 mRNA levels were related to the agonal state. The aim of our study was to demonstrate the presence of Hsp70--immunoreactive protein in brain of controls, patients with AD and Down Syndrome (DS) in individual brain regions. The rationale for the study was to test the hypothesis that expression of Hsp70, a protein involved in apoptosis would be altered in brain of these patients with neurodegenerative disorders where (neuronal) apoptosis is a hallmark of the disease. Brain immunoreactive-Hsp70--protein (Hsp70) was determined by Western blotting using specific monoclonal antibody in five different brain regions (frontal, parietal, occipital, temporal cortex and cerebellum) from controls, DS and AD patients. Hsp70 expression was significantly increased in temporal cortex of patients with AD (arbitrary units: means +/- SD; 0.35 +/- 0.49 for controls, 0.97 +/- 0.70 for DS patients, 1.16 +/- 0.56 for AD patients). In frontal and parietal cortex from DS patients, there was a strong correlation between Hsp70 levels and the length of post-mortem interval (r = 0.95, P < 0.01 and r = 0.82, P < 0.021). PMID- 10666687 TI - Increased levels of 14-3-3 gamma and epsilon proteins in brain of patients with Alzheimer's disease and Down syndrome. AB - The 14-3-3 family consists of homo- and heterodimeric proteins representing a novel type of "adaptor proteins" modulating the interaction between components of signal transduction pathways. 14-3-3 isoforms interact with phosphoserine motifs on many proteins as kinases, phosphatases, apoptosis related proteins etc. Performing protein mapping by 2D electrophoresis in human brain we identified two isoforms, 14-3-3 gamma and epsilon and decided to determine these two multifunctional proteins in several brain regions of aged patients with Alzheimer's disease (AD) and Down Syndrome (DS) with AD neuropathology in comparison with control brains. 14-3-3 gamma and 14-3-3 epsilon proteins were increased in several brain regions of AD and DS patients. These changes may contribute to the complex pathomechanisms of AD and AD in DS, evolving inevitably from the fourth decade of life. Deranged 14-3-3 isoforms gamma and epsilon may reflect impaired signaling and/or apoptosis in the brain as several kinases (protein kinase C, Ras, mitogen-activated kinase MEK) involved in signaling and apoptotic factors as bcl-2-related proteins BAD and BAG-1 are binding to 14-3-3 motifs. PMID- 10666688 TI - Application of Alu-splice PCR on chromosome 21: DSCR1 and Intersectin. AB - Down syndrome (DS) is a major cause of mental retardation and congenital heart defects, with an overall incidence of one in 700 live births. DS is caused by increases in the amounts of a number of normal gene products, the exact number and identity of which are presently unknown. Elucidating the molecular basis of DS relies on the identification of the gene products whose augmentation by 50% or more causes symptoms of the disease. With the aim of contributing to the transcriptional map of human chromosome 21 and to identify new genes with potential involvement in DS, we developed a technique to isolate expressed sequences called Alu-splice PCR, which is very simple to perform and is independent of gene expression patterns. Putative exons are PCR amplified in genomic DNA by virtue of their proximity to Alu repeats using primers designed from splice-site consensus sequences in combination with specific Alu repeat primers. The Alu repeats, which are repetitive DNA elements found exclusively and at high frequency in the genomes of primates, impart the human specificity to the method. The splice-site consensus sequences were used to direct primers to exon boundaries. Using the Alu-splice technique, we have identified at least three new genes. We trapped an exon of DSCR1 (Down Syndrome Candidate Region 1) and two different exons of a gene called human Intersectin (ITSN). Presently, we are working with another novel trapped exon to identify the corresponding gene. The major advantage of Alu-splice PCR is that the technique can be readily established in any laboratory which has the basic facilities for molecular biology because no specialised materials or expertise is required. PMID- 10666689 TI - Overexpression of DNAse I in brain of patients with Down syndrome. AB - Human DNAse I (EC 3.1.21.1) is an enzyme most probably involved in apoptotic processes. Splicing of the DNAse I primary transcript in normal and apoptotic cells into up to 20 splicing forms and the recent description of a different family of caspase-activated DNAses, hampered studies on the role of DNAse I in apoptosis research. Performing gene hunting in fetal brain of patients with DS we found a sequence with 100% homology to DNAse I and this formed the Rationale for studies in adult DS brain. It was therefore the aim of the study to evaluate DNAse I-mRNA steady state levels in DS brain using adult brain without brain pathologies and Alzheimer's Disease (AD) brain as control, in order to rule out that DNAse I--overexpression may not be specific for DS but rather reflecting apoptosis per se, a hallmark of both disorders. Determination of DNAse I-mRNA steady state levels was carried out by a blotting method in frontal, parietal, temporal occipital lobe and cerebellum. We found significantly increased DNAse I transcripts in brain of DS and AD both, when normalized versus the house-keeping gene beta actin or total RNA. We demonstrate the significant increase of DNAse I- transcript in the pathogenesis of DS and AD suggesting a role for this enzyme in the apoptotic process known to occur in both disorders. We are now going to carry out protein and enzyme activity levels in our laboratory to confirm our findings at the transcriptional level. PMID- 10666690 TI - [The hyoid bone and its preservation in the framework of supraglottic partial resection of the larynx]. AB - BACKGROUND: Whether to remove the hyoid bone as part of the horizontal partial laryngectomy is a question which is discussed controversially. Pathohistological examination of the region involved should help to answer this question. MATERIAL AND METHODS: Regarding the involvement of the hyoid bone, 71 surgical specimens of supraglottic carcinoma invading the pre-epiglottic space were studied by whole organ serial sections. RESULTS: Only four specimens demonstrated mandatory resection of the hyoid bone because of its proximity to the advancing border of the tumor. In these cases, large tumor masses were palpable directly under the thyrohyoid membrane after the strap muscles had been dissected during the operation. DISCUSSION: Except for these rare cases in which the tumor was already palpable from outside the larynx, preservation of the hyoid bone during horizontal supraglottic laryngectomy is oncologically safe. It provides a more secure closure of the pharyngotomy and promotes earlier return of the swallowing function because of the maintenance of the normal position of the larynx and pharynx. PMID- 10666691 TI - [Pathomorphological aspects of transoral resection of hypopharyngeal carcinoma with preservation of the larynx. Patient selection, treatment results]. AB - BACKGROUND: In primary surgery of hypopharyngeal cancer, transcervical resection was chosen in order to preserve the larynx. This treatment produces good oncological results but also a high degree of morbidity so that in recent years transoral resection has been recommended. For wider application of this method it is very important to apply clearly defined criteria for selection of patients. MATERIAL AND METHODS: To define the criteria for selection for transoral microsurgical resection, we analyzed step serial sections of 33 whole organ specimens of hypopharyngeal squamous cell cancer (SCC) after primary radical surgery, mostly carcinoma of the piriform sinus. Criteria concerning the primary and the involvement of the neck nodes were differentiated. Twenty of 84 patients with hypopharyngeal cancer of different stages were treated by transoral resection and delayed neck dissection between 1994 and 1996. Most of the patients were irradiated postoperatively because of neck metastases. The therapeutic results after a minimum period of 24 months follow-up is listed according to Kaplan-Meyer. RESULTS: Three types were defined according to their site, growth, and spread into the larynx: Type I comprises limited exophytic, highly differentiated SCC with a minor tendency for metastasis originating in the upper half of the sinus. These tumors are well suitable for transoral resection. Type II includes tumors spreading superficially without deeper invasion of the larynx, especially of the laryngeal framework. These can be totally resected and the larynx preserved in spite of extended metastasis. Type III, the most frequent type, grows with ulceration and deeply infiltrates into the larynx and the neck. These tumors cannot be resected transorally. Primary radical resection is indicated. Up to 25% of all hypopharyngeal SCC could be treated by transoral resection, mostly with delayed neck dissection and postoperative irradiation. Functional results were excellent in all cases. Eating, voice, and air passage were normal. Oncological results with 80% disease free five-year survival rate were very good. Three patients died because of recurrences in the neck, only one because of a recurrence in the larynx. The rate of patients with a second primary, however, was extremely high (50%). CONCLUSIONS: By strictly following the pathological and clinical criteria for selection, about 25% of the SCC of the hypopharynx can be treated by transoral resection combined with neck dissection and postoperative irradiation with good oncological and excellent functional results, preserving the larynx without endangering the life of the patients. PMID- 10666692 TI - [Studies on significance of sentinel lymphadenectomy in pharyngeal and laryngeal carcinoma]. AB - BACKGROUND: Management of the suspected N0-neck (sonography and CT) in squamous cell carcinoma (SCC) of the head and neck is discussed controversially. The question arises whether the sentinel node (SN) concept as it is performed in different areas of clinical oncology is applicable to ear, nose, and throat medicine. METHODS: Nine male patients with SCC were studied (4 oropharynx, 2 hypopharynx, and 3 larynx) in whom different lymph node status was diagnosed clinically (5 x N0, 2 x N1, 2 x N2c). After intraoperative scintillation probe detection, the histological examination of the SN with neck dissection (ND) specimen followed. RESULTS: In 7 of 9 cases SN detection was successful. In 4 of 5 cases of clinical N0 status, SN, and ND specimens were free of tumor histologically, while in one patient radiolabel-identified SN showed tumor cells in histological examination. In 2 patients with clinical N1 neck, SN, and ND were histologically tumor-free in one patient and contained one single tumor metastasis located in the SN in the other patient. In 2 patients with clinically and histologically proven N2c neck, lymph nodes located in regions II and III showed metastasis including capsular rupture. In both cases no lymph node radioactivity was detectable during the operation. CONCLUSIONS: The results suggest that sentinel lymphonodectomy may be suited for ear, nose, and throat medicine. Before it is applied to clinical practice, further problems must be resolved. These include the short distance between the primary injection side and lymph nodes and the influence of intranodal tumor metastasis on the uptake of the radiolabeled tracer. PMID- 10666693 TI - [Multiple carcinomas in the upper aerodigestive tract. 1. Oral cavity and oropharynx]. AB - BACKGROUND: During the last years an absolute increase of tumour incidence of squamous cell carcinoma as well as an increase in the occurrence of synchronous and metachronous multiple primaries in the upper aerodigestive tract can be observed. This study analyses the so-called "multi-centric cancerization" in patients with primary carcinoma of the oral cavity and the oropharynx. METHODS: During 2 observation periods of 5 years each, from 1985 to 1994, we retrospectively analyzed 235 patients with squamous cell carcinoma of the oral cavity and 232 patients with tumour localisation in the oropharynx. Besides age, sex, tumour localization, TNM-stage and grading, the risk factors tobacco and alcohol were added as causes for the development of multiple primaries. RESULTS: In the primary localisation of the oral cavity synchronous and metachronous double tumours increased from 7% to 17% besides the absolute increase in tumour incidence. In the oropharynx a total increase of second carcinomas from 3% to 20% was found. At the same time a growing abuse of tobacco and alcohol could be observed. CONCLUSIONS: Panendoscopy during pre-therapeutical staging should be obligatory. Regular oncological controls are mandatory, especially for younger patients with smaller tumours and good prognosis, but a high risk of second primaries. In the long run, prevention has to play a decisive role in the fight against second primary tumors of the upper aerodigestive tract. Possible improvements of early diagnosis, genetical examinations, information campaigns as well as research of carcinogenic environmental pollutants are of foremost interest to the clinician. PMID- 10666694 TI - [Mutagen sensitivity of patients with laryngeal and oropharyngeal carcinoma]. AB - BACKGROUND: Carcinogenesis in the larynx and oropharynx is often associated with excessive exposure to tobacco smoke and alcohol. However, attention is increasingly being focused on genetically determined mutagen sensitivities and on the mutagenic impact of xenobiotics. The purpose of this study was to evaluate the genotoxicity of phthalates (plasticizers widely used in synthetic materials), as well as nitrosamines and polycyclic aromatic carbohydrates, on laryngeal and oropharyngeal epithelia and peripheral lymphocytes of patients with laryngeal and oropharyngeal carcinomas. METHODS: The comet assay was used to detect induced DNA strand breaks. Macroscopically healthy supraglottic and oropharyngeal epithelia of patients with laryngeal and oropharyngeal tumors, respectively, and lymphocytes were investigated with dibutyl phthalate (DBP), diisobutylphthalate (DiBP). N'nitrosodiethylamine (NDELA), and benzo[a]pyrene (BaP). The Olive Tail Moment (OTM) was used to quantify genotoxicity. RESULTS: For the first time, the genotoxicity of DBP and DiBP was demonstrated in laryngeal and oropharyngeal epithelia, as well as in peripheral lymphocytes, of patients suffering from laryngeal and oropharyngeal carcinomas. OTM levels for NDELA were higher than for phthalates; levels for BaP were lower. Testing of lymphocytes and mucosa showed no significant differences among the various substances. CONCLUSIONS: Phthalates show a genotoxic impact on epithelia of tumor patients. OTM levels were higher than in nasal and oropharyngeal mucosa of healthy donors in results reported earlier. Thus, specific susceptibilities to these xenobiotics need to be discussed. No such effect was demonstrated for NDELA and BaP. In tumor patients, no significant differences could be shown in mutagenic sensitivities in mucosal cells and lymphocytes. PMID- 10666695 TI - [Endoscopic imaging techniques in the diagnosis of laryngeal carcinoma and its precursor lesions]. AB - BACKGROUND: In order to improve preoperative diagnostic work-up in treatment of patients with laryngeal cancer and its precursor lesions additional endoscopical imaging techniques have been developed: 1. Autofluorescence endoscopy; 2. Contact endoscopy; 3. Endoscopic high-frequency ultrasound. These imaging techniques are used during microlaryngoscopy to get further information about tumor extension and differentiation. This paper describes the diagnostic potential of these imaging techniques in the evaluation of cancerous lesions of the larynx. MATERIAL AND METHODS: Patients in different stages of laryngeal dysplasia, carcinoma in situ and laryngeal cancer were examined by means of the previous mentioned imaging techniques during microlaryngoscopy (Autofluorescence endoscopy [n = 38], contact endoscopy [n = 323], endoscopic high-frequency ultrasound [n = 60]) and the results were compared to pathohistological findings. In autofluorescence endoscopy cancerous mucosa was illuminated using blue filtered light (380-460 nm) to obtain autofluorescence for optical demarcation of the lesion. Contact endoscopy was performed after staining of the laryngeal mucosa with methylene blue (1%). Two different endoscopes with 60 x and 150 x magnification were used. In both techniques a video image was achieved by using a xenon light source and a special video camera to register autofluorescence. The endoscopical high frequency ultrasound examination was performed after flooding the larynx with 0.9% saline. Newly developed ultrasound catheters with frequencies between 10 to 20 MHz were inserted in the laryngeal lumen and moved in a standardized pattern during the examination. RESULTS: During the autofluorescence examination of the endolaryngeal mucosa precancerous and cancerous lesions showed a red to violet fluorescence outlined against the light green autofluorescence of the normal mucosa. Hyperplastic hyperkeratotic epithelium revealed a higher intensity of light green or even whitish autofluorescence compared to normal mucosa autofluorescence. After staining the vocal cords with methylene blue, it was possible to observe the cells, nuclei and cytoplasm of the laryngeal mucosa and their different grades of abnormality using the specially developed contact endoscopes. Endoscopic high-frequency ultrasound (10 to 20 MHz) was able to measure the vertical extension of laryngeal carcinomas bigger than 3 mm in size. The involvement of the thyroid cartilage or the anterior commissure could be visualized. Preoperatively, the critical T2 stage could be evaluated more precisely. In precancerous lesions and microinvasive cancer ultrasound added no additional Information to the microlaryngoscopical picture. CONCLUSION: Autofluorescence, contact endoscopy as well as endoscopic high-frequency ultrasound are promising new imaging techniques supplementing microlaryngoscopy: autofluorescence as well as contact endoscopy are suitable to differentiate dysplasia, carcinoma in situ, microinvasive lesions as well as the evaluation of tumorous margins, while high-frequency ultrasound improves the assessment of tumorous infiltration into the depth of the larynx. These imaging techniques enable the laryngologist to perform a more accurate diagnostic work-up in the assessment of laryngeal cancer and its precursor lesions. PMID- 10666696 TI - [Long-term results of homograft reconstruction of the trachea in childhood]. AB - BACKGROUND: The treatment of long-segment tracheal stenosis in children is a biological and operative problem. The introduction of preserved allografts established new possibilities for functional tracheal reconstruction. The development of tracheal dimensions in the course of growing is often discussed. PATIENTS AND METHODS: Since 1983 preserved allografts were used in 20 children with acquired tracheal stenosis. The children ranged in age from 1 to 13 years (average: 7.2 years) when the operation was performed. The children were examined postoperatively at adequate intervals. Radiologic measurements of the tracheal dimensions were performed in some of the patients. RESULTS: Today the treatment of all of these children is complete. Some of them are now adults. None of the children demonstrated breathing problems at the time of the end of treatment or after a variable following-up period ranging from 18 months to 14 years. There was no endoscopic or radiographic evidence of constriction in the growing tracheas. Radiographic measurement of the reconstructed tracheas showed an age adequate growth in length and a small growth in diameter. CONCLUSION: Functional tracheal reconstruction for long-segment tracheal stenosis can be achieved by implantation of preserved allografts even in the early childhood. PMID- 10666697 TI - [Combined transseptal-transnasal surgery of unilateral choanal atresia without using stents]. AB - BACKGROUND: This is a report of a surgical procedure with which stenting after surgery of unilateral choanal atresia can be avoided. METHOD: We start transseptally with a hemitransfixion incision. The vomer and the bony atresia plate are dissected submucosally and then resected with punches and a diamond burr, whilst the nasal and the pharyngeal mucosa of the atresia is preserved completely. The next step is to cut transnasally a door-like flap out of the nasal blade of the mucosa of the atresia, that is based laterally and with which the lateral raw surface of the new choana will be covered. Now a medially based door-like flap is cut out of the up to now intact pharyngeal blade of the atresia, with which the medial wound surface is to be covered. The flaps are fixed with fibrin glue and with a nasal tamponade for two days, as after septoplasty. It is not necessary to insert stents anymore. RESULTS: Between 1995 until 1998 we treated three female and two male patients using this method. Their ages were 6, 17, 17, 20 and 35 years and the postoperative observation lasted 2, 4, 1, 3 and 3 years. A restenosis was not seen in any of the cases. CONCLUSION: Because of these results, it is recommended to treat unilateral choanal atresias of adults and children older than five years by using a combined transseptal transnasal procedure. By using the described surgical technique postoperative stenting can be avoided. PMID- 10666698 TI - [Current aspects of hearing aid fitting: technique, goals and testing]. PMID- 10666699 TI - [The interesting case No. 31. Eyelid inflammation in childhood]. PMID- 10666700 TI - [100 years of the Otto Korner ENT Clinic, Rostock. The merits of Christian Lemcke and Otto Korner in the development of ENT specialty]. AB - BACKGROUND: The first special ENT hospital in Germany and northern Europe, the Department of Otorhinolaryngology, Head- and Neck-Surgery "Otto Korner" in Rostock is celebrating its 100th anniversary on October 25, 1999. METHODS: There is a presentation on the efforts in the ENT-specialty from the beginning considering the special situation at the university of Rostock. RESULTS: The efforts of Christian Lemcke (1850-1894), who associated the three sections Otology, Laryngology and Rhinology are appreciated, Otto Korner (1859-1935) continued this development. He achieved the building of a special ENT-hospital by his excellent knowledge and surgical skills. The opening date was October 25, 1899, Korner became the first full professor in Otology and Laryngology in Germany in 1901. His "Textbook Of Otology And It's Bordering Specialties" was published in 1906. As one of the first, he supported the independence of ENT medicine as a separate field in the course of medicine. CONCLUSION: Today, Otto Korner's claims are more relevant than ever before, because the today's students in their practical course of medicine and even young doctors at the hospitals obviously show a lack of knowledge in the ENT-field. The current "multiple choice" type of examinations could be one of the main reasons. PMID- 10666701 TI - [Surgical measures in pharyngo-esophageal dysphagia and chronic aspiration]. PMID- 10666702 TI - Population bottlenecks and Pleistocene human evolution. AB - We review the anatomical and archaeological evidence for an early population bottleneck in humans and bracket the time when it could have occurred. We outline the subsequent demographic changes that the archaeological evidence of range expansions and contractions address, and we examine how inbreeding effective population size provides an alternative view of past population size change. This addresses the question of other, more recent, population size bottlenecks, and we review nonrecombining and recombining genetic systems that may reflect them. We examine how these genetic data constrain the possibility of significant population size bottlenecks (i.e., of sufficiently small size and/or long duration to minimize genetic variation in autosomal and haploid systems) at several different critical times in human history. Different constraints appear in nonrecombining and recombining systems, and among the autosomal loci most are incompatible with any Pleistocene population size expansions. Microsatellite data seem to show Pleistocene population size expansions, but in aggregate they are difficult to interpret because different microsatellite studies do not show the same expansion. The archaeological data are only compatible with a few of these analyses, most prominently with data from Alu elements, and we use these facts to question whether the view of the past from analysis of inbreeding effective population size is valid. Finally, we examine the issue of whether inbreeding effective population size provides any reasonable measure of the actual past size of the human species. We contend that if the evidence of a population size bottleneck early in the evolution of our lineage is accepted, most genetic data either lack the resolution to address subsequent changes in the human population or do not meet the assumptions required to do so validly. It is our conclusion that, at the moment, genetic data cannot disprove a simple model of exponential population growth following a bottleneck 2 MYA at the origin of our lineage and extending through the Pleistocene. Archaeological and paleontological data indicate that this model is too oversimplified to be an accurate reflection of detailed population history, and therefore we find that genetic data lack the resolution to validly reflect many details of Pleistocene human population change. However, there is one detail that these data are sufficient to address. Both genetic and anthropological data are incompatible with the hypothesis of a recent population size bottleneck. Such an event would be expected to leave a significant mark across numerous genetic loci and observable anatomical traits, but while some subsets of data are compatible with a recent population size bottleneck, there is no consistently expressed effect that can be found across the range where it should appear, and this absence disproves the hypothesis. PMID- 10666703 TI - Evidence from beta-tubulin phylogeny that microsporidia evolved from within the fungi. AB - Microsporidia are obligate intracellular parasites that were thought to be an ancient eukaryotic lineage based on molecular phylogenies using ribosomal RNA and translation elongation factors. However, this ancient origin of microsporidia has been contested recently, as several other molecular phylogenies suggest that microsporidia are closely related to fungi. Most of the protein trees that place microsporidia with fungi are not well sampled, however, and it is impossible to resolve whether microsporidia evolved from a fungus or from a protistan relative of fungi. We have sequenced beta-tubulins from 3 microsporidia, 4 chytrid fungi, and 12 zygomycete fungi, expanding the representation of beta-tubulin to include all four fungal divisions and a wide diversity of microsporidia. In phylogenetic trees including these new sequences, the overall topology of the fungal beta tubulins generally matched the expected relationships among the four fungal divisions, although the zygomycetes were polyphyletic in some analyses. The microsporidia consistently fell within this fungal diversification, and not as a sister group to fungi. Overall, beta-tubulin phylogeny suggests that microsporidia evolved from a fungus sometime after the divergence of chytrids. We also found that chytrid alpha- and beta-tubulins are much less divergent than are tubulins from other fungi or microsporidia. In trees in which the only fungal representatives were the chytrids, microsporidia still branched with fungi (i.e., with chytrids), suggesting that the affiliation between microsporidian and fungal tubulins is not an artifact of long-branch attraction. PMID- 10666704 TI - Estimating synonymous and nonsynonymous substitution rates under realistic evolutionary models. AB - Approximate methods for estimating the numbers of synonymous and nonsynonymous substitutions between two DNA sequences involve three steps: counting of synonymous and nonsynonymous sites in the two sequences, counting of synonymous and nonsynonymous differences between the two sequences, and correcting for multiple substitutions at the same site. We examine complexities involved in those steps and propose a new approximate method that takes into account two major features of DNA sequence evolution: transition/transversion rate bias and base/codon frequency bias. We compare the new method with maximum likelihood, as well as several other approximate methods, by examining infinitely long sequences, performing computer simulations, and analyzing a real data set. The results suggest that when there are transition/transversion rate biases and base/codon frequency biases, previously described approximate methods for estimating the nonsynonymous/synonymous rate ratio may involve serious biases, and the bias can be both positive and negative. The new method is, in general, superior to earlier approximate methods and may be useful for analyzing large data sets, although maximum likelihood appears to always be the method of choice. PMID- 10666705 TI - A diverse population of introns in the nuclear ribosomal genes of ericoid mycorrhizal fungi includes elements with sequence similarity to endonuclease coding genes. AB - Ericoid mycorrhizal fungi form symbioses with the roots of members of the Ericales. Although only two genera have been identified in culture, the taxonomic diversity of ericoid symbionts is certainly wider. Genetic variation among 40 ericoid fungal isolates was investigated in this study. PCR amplification of the nuclear small-subunit ribosomal DNA (SSU rDNA) and of the internal transcribed spacer (ITS), followed by sequencing, led to the discovery of DNA insertions of various sizes in the SSU rDNA of most isolates. They reached sizes of almost 1,800 bp and occurred in up to five different insertion sites. Their positions and sizes were generally correlated with morphological and ITS-RFLP grouping of the isolates, although some insertions were found to be optional among isolates of the same species, and insertions were not always present in all SSU rDNA repeats within an isolate. Most insertions were identified as typical group I introns, possessing the conserved motifs characteristic of this group. However, other insertions lack these motifs and form a distinct group that includes other fungal ribosomal introns. Alignments with almost 70 additional sequences from fungal nuclear SSU rDNA introns indicate that introns inserted at the same site along the rDNA gene are generally homologous, but they also suggest the possibility of some horizontal transfers. Two of the ericoid fungal introns showed strong homology with a conserved motif found in endonuclease genes from nuclear rDNA introns. PMID- 10666706 TI - The phylogenetic position of the Talpidae within eutheria based on analysis of complete mitochondrial sequences. AB - The complete mitochondrial (mt) genome of the mole Talpa europaea was sequenced and included in phylogenetic analyses together with another lipotyphlan (insectivore) species, the hedgehog Erinaceus europaeus, and 22 other eutherian species plus three outgroup taxa (two marsupials and a monotreme). The phylogenetic analyses reconstructed a sister group relationship between the mole and fruit bat Artibeus jamaicensis (order Chiroptera). The Talpa/Artibeus clade constitutes a sister clade of the cetferungulates, a clade including Cetacea, Artiodactyla, Perissodactyla, and Carnivora. A monophyletic relationship between the hedgehog and the mole was significantly rejected by maximum parsimony and maximum likelihood. Consistent with current systematic schemes, analyses of complete cytochrome b genes including the shrew Sorex araneus (family Soricidae) revealed a close relationship between Talpidae and Soricidae. The analyses of complete mtDNAs, along with the findings of other insectivore studies, challenge the maintenance of the order Lipotyphla as a taxonomic unit and support the elevation of the Soricomorpha (with the families Talpidae and Soricidae and possibly also the Solenodontidae and Tenrecidae) to the level of an order, as previously proposed in some morphological studies. PMID- 10666707 TI - Determinants of substitution rates in mammalian genes: expression pattern affects selection intensity but not mutation rate. AB - To determine whether gene expression patterns affect mutation rates and/or selection intensity in mammalian genes, we studied the relationships between substitution rates and tissue distribution of gene expression. For this purpose, we analyzed 2,400 human/rodent and 834 mouse/rat orthologous genes, and we measured (using expressed sequence tag data) their expression patterns in 19 tissues from three development states. We show that substitution rates at nonsynonymous sites are strongly negatively correlated with tissue distribution breadth: almost threefold lower in ubiquitous than in tissue-specific genes. Nonsynonymous substitution rates also vary considerably according to the tissues: the average rate is twofold lower in brain-, muscle-, retina- and neuron-specific genes than in lymphocyte-, lung-, and liver-specific genes. Interestingly, 5' and 3' untranslated regions (UTRs) show exactly the same trend. These results demonstrate that the expression pattern is an essential factor in determining the selective pressure on functional sites in both coding and noncoding regions. Conversely, silent substitution rates do not vary with expression pattern, even in ubiquitously expressed genes. This latter result thus suggests that synonymous codon usage is not constrained by selection in mammals. Furthermore, this result also indicates that there is no reduction of mutation rates in genes expressed in the germ line, contrary to what had been hypothesized based on the fact that transcribed DNA is more efficiently repaired than nontranscribed DNA. PMID- 10666708 TI - Molecular evolution of the Paramyxoviridae and Rhabdoviridae multiple-protein encoding P gene. AB - Presented here is an analysis of the molecular evolutionary dynamics of the P gene among 76 representative sequences of the Paramyxoviridae and Rhabdoviridae RNA virus families. In a number of Paramyxoviridae taxa, as well as in vesicular stomatitis viruses of the Rhabdoviridae, the P gene encodes multiple proteins from a single genomic RNA sequence. These products include the phosphoprotein (P), as well as the C and V proteins. The complexity of the P gene makes it an intriguing locus to study from an evolutionary perspective. Amino acid sequence alignments of the proteins encoded at the P and N loci were used in independent phylogenetic reconstructions of the Paramyxoviridae and Rhabdoviridae families. P gene-coding capacities were mapped onto the Paramyxoviridae phylogeny, and the most parsimonious path of multiple-coding-capacity evolution was determined. Levels of amino acid variation for Paramyxoviridae and Rhabdoviridae P-gene encoded products were also analyzed. Proteins encoded in overlapping reading frames from the same nucleotides have different levels of amino acid variation. The nucleotide architecture that underlies the amino acid variation was determined in order to evaluate the role of selection in the evolution of the P gene overlapping reading frames. In every case, the evolution of one of the proteins encoded in the overlapping reading frames has been constrained by negative selection while the other has evolved more rapidly. The integrity of the overlapping reading frame that represents a derived state is generally maintained at the expense of the ancestral reading frame encoded by the same nucleotides. The evolution of such multicoding sequences is likely a response by RNA viruses to selective pressure to maximize genomic information content while maintaining small genome size. The ability to evolve such a complex genomic strategy is intimately related to the dynamics of the viral quasispecies, which allow enhanced exploration of the adaptive landscape. PMID- 10666709 TI - Mitochondrial genomes of Galathealinum, Helobdella, and Platynereis: sequence and gene arrangement comparisons indicate that Pogonophora is not a phylum and Annelida and Arthropoda are not sister taxa. AB - We report a contiguous region of more than half (> 7,500 nt) of the mitochondrial genomes for Platynereis dumerii (Annelida: Polychaeta), Helobdella robusta (Annelida: Hirudinida), and Galathealinum brachiosum (Pogonophora: Perviata). The relative arrangements of all 22 genes identified for Helobdella and Galathealinum are identical to one another and to their arrangements in the mtDNA of the previously studied oligochaete annelid Lumbricus. In contrast, Platynereis differs from these taxa in the positions of several tRNA genes and in having two additional tRNA genes (trnC and trnM) and a large noncoding sequence in this region. Comparisons of relative gene arrangements and of the nucleotide and inferred amino acid sequences among these and other published taxa provide strong support for an annelid-mollusk clade that excludes arthropods, and for the inclusion of pogonophorans within Annelida, rather than giving them separate phylum status. Gene arrangement comparisons include the first use of a recently described method on previously unpublished data. Although a variety of alternative initiation codons are typically used by mitochondrial protein encoding genes, ATG appears to be the initiator for all but one reported here. The large noncoding region (1,091 nt) identified in Platynereis has no significant sequence similarity to the noncoding region of Lumbricus, although each contains runs of TA dinucleotides and of homopolymers, which could potentially serve as signaling elements. There is strong bias for synonymous codon usage in Helobdella and especially in Galathealinum. In this latter taxon, 5 codons are completely unused, 13 are used three or fewer times, and G appears at third codon positions in only 26 of the 2,236 codons. Nucleotide composition bias appears to influence amino acid composition of the proteins. PMID- 10666710 TI - Mitochondrial genomic rearrangements in songbirds. AB - The organization of the mitochondrial genome is generally very conserved among vertebrates. Because of this, examination of the rare rearrangements which do occur has been suggested as offering a powerful alternative to phylogenetic analyses of mitochondrial DNA sequences. Here, we report on an avian mitochondrial rearrangement in a group of oscine passerines (warblers of the genus Phylloscopus). This rearrangement is identical to the mitochondrial organization recently identified in representatives of four orders of birds, including subsoscine Passeriformes. The rearrangement involves the movement of three genes (tRNA(Pro), NADH6, and tRNA(Glu)) from their normal position in birds between tRNA(Thr) and the control region (CR), to a new location between the CR and a novel, supposedly noncoding (NC), region. Our results suggest that this derived arrangement cannot be used to distinguish between suboscine and oscine passerines, as it has multiple origins both within Passeriformes and within birds as a whole. We found short stretches of DNA with high degrees of similarity between the CR and each NC region, respectively, all of which could be located in the same area of the CR. This suggests that the CR and the NC region are homologous and that the mechanism behind this mitochondrial rearrangement is a tandem duplication followed by multiple deletions. However, the similarities between the control and NC regions of each species were less pronounced than those between the control or NC regions from the different species, supporting the hypothesis of a single basal rearrangement in the Phylloscopus warblers. PMID- 10666711 TI - A phylogenetic analysis of the lipocalin protein family. AB - The lipocalins are a family of extracellular proteins that bind and transport small hydrophobic molecules. They are found in eubacteria and a great variety of eukaryotic cells, in which they play diverse physiological roles. We report here the detection of two new eukaryotic lipocalins and a phylogenetic analysis of 113 lipocalin family members performed with maximum-likelihood and parsimony methods on their amino acid sequences. Lipocalins segregate into 13 monophyletic clades, some of which are grouped in well-supported superclades. An examination of the G + C content of the bacterial lipocalin genes and the detection of four new conceptual lipocalins in other eubacterial species argue against a recent horizontal transfer as the origin of prokaryotic lipocalins. Therefore, we rooted our lipocalin tree using the clade containing the prokaryotic lipocalins. The topology of the rooted lipocalin tree is in general agreement with the currently accepted view of the organismal phylogeny of arthropods and chordates. The rooted tree allows us to assign polarity to character changes and suggests a plausible scenario for the evolution of important lipocalin properties. More recently evolved lipocalins tend to (1) show greater rates of amino acid substitutions, (2) have more flexible protein structures, (3) bind smaller hydrophobic ligands, and (4) increase the efficiency of their ligand-binding contacts. Finally, we found that the family of fatty-acid-binding proteins originated from the more derived lipocalins and therefore cannot be considered a sister group of the lipocalin family. PMID- 10666712 TI - Rh gene evolution in primates: study of intron sequences. AB - By amplification and sequencing of RH gene intron 4 of various primates we demonstrate that an Alu-Sx-like element has been inserted in the RH gene of the common ancestor of humans, apes, Old World monkeys, and New World monkeys. The study of mouse and lemur intron 4 sequences allowed us to precisely define the insertion point of the Alu-Sx element in intron 4 of the RH gene ancestor common to Anthropoidea. Like humans, chimpanzees and gorillas possess two types of RH intron 4, characterized by the presence (human RHCE and ape RHCE-like genes) or absence (human RHD and ape RHD-like genes) of the Alu-Sx element. This led us to conclude that in the RH common ancestor of humans, chimpanzees, and gorillas, a duplication of the common ancestor gene gave rise to two genes, one differing from the other by a 654-bp deletion encompassing an Alu-Sx element. Moreover, most of chimpanzees and some gorillas posses two types of RHD-like intron 4. The introns 4 of type 1 have a length similar to that of human RHD intron 4, whereas introns 4 of type 2 display an insertion of 12 bp. The latest insertion was not found in the human genome (72 individuals tested). The study of RH intron 3 length polymorphism confirmed that, like humans, chimpanzees and gorillas possess two types of intron 3, with the RHD-type intron 3 being 289 bases shorter than the RHCE intron 3. By amplification and sequencing of regions encompassing introns 3 and 4, we demonstrated that chimpanzee and gorilla RH-like genes displayed associations of introns 3 and 4 distinct to those found in man. Altogether, the results demonstrate that, as in humans, chimpanzee and gorilla RH genes experienced intergenic exchanges. PMID- 10666713 TI - Pattern of morphological diversification in the Leptocarabus ground beetles (Coleoptera: Carabidae) as deduced from mitochondrial ND5 gene and nuclear 28S rDNA sequences. AB - Most of the mitochondrial NADH dehydrogenase subunit 5 (ND5) gene and a part of nuclear 28S ribosomal RNA gene were sequenced for 14 species of ground beetles belonging to the genus Leptocarabus. In both the ND5 and the 28S rDNA phylogenetic trees of Leptocarabus, three major lineages were recognized: (1) L. marcilhaci/L. yokoael/Leptocarabus sp. from China, (2) L. koreanus/L. truncaticollis/L. seishinensis/L. semiopacus/L. canaliculatus/L. kurilensis from the northern Eurasian continent including Korea and Hokkaido, Japan, and (3) all of the Japanese species except L. kurilensis. Clustering of the species in the trees is largely linked to their geographic distribution and does not correlate with morphological characters. The species belonging to different species groups are clustered in the same lineages, and those in the same species group are scattered among the different lineages. One of the possible interpretations of the present results would be that morphological transformations independently took place in the different lineages, sometimes with accompanying parallel morphological evolution, resulting in the occurrence of the morphological species belonging to the same species group (= type) in the different lineages. PMID- 10666714 TI - The evolution of vertebrate antigen receptors: a phylogenetic approach. AB - Classical T cells, those with alpha beta T-cell receptors (TCRs), are an important component of the dominant paradigm for self-nonself immune recognition in vertebrates. alpha beta T cells recognize foreign peptide antigens when they are bound to MHC molecules on the surfaces of antigen-presenting cells. gamma delta T cells bear a similar receptor, and it is often assumed that these T cells also require specialized antigen-presenting molecules for immune recognition, which we term "indirect antigen recognition." B-cell receptors, or immunoglobulins, bind directly to antigens without the help of a specialized antigen-presenting molecule. Phylogenetically, it has been assumed that T-cell receptors and the genes that encode them are a monophyletic group, and that "indirect" antigen recognition evolved before the split into two types of TCR. Recently, however, it has been proposed that gamma delta-TCRs bind directly to antigens, as do immunoglobulins (Ig's). This calls into question the null hypothesis that indirect antigen recognition is a common characteristic of TCRs and, by extension, the hypothesis that all TCR gene sequences form a monophyletic group. To determine whether alternative explanations for antigen recognition and other historical relationships among TCR genes might be possible, we performed phylogenetic analyses on amino acid sequences of the constant and variable regions which encode the basic subunits of TCR and Ig molecules. We used both maximum-parsimony and genetic distance-based methods and could find no strong support for the hypothesis of TCR monophyly. Analyses of the constant region suggest that TCR gamma or delta sequences are the most ancient, implying that the ancestral immune cell was like a modern gamma delta T cell. From this gamma delta like ancestor arose alpha beta T cells and B cells, implying that indirect antigen recognition is indeed a derived property of alpha beta-TCRs. Analyses of the variable regions are complicated by strong selection on antigen-binding sequences, but imply that direct antigen binding is the ancestral condition. PMID- 10666715 TI - A comparison of estimators of the population recombination rate. AB - Three new estimators of the population recombination rate C = 4Nr are introduced. These estimators summarize the data using the number of distinct haplotypes and the estimated minimum number of recombination events, then calculate the value of C that maximizes the likelihood of obtaining the summarized data. They are compared with a number of previously proposed estimators of the recombination rate. One of the newly proposed estimators is generally better than the others for the parameter values considered here, while the three programs that calculate maximum-likelihood estimates give conflicting results. PMID- 10666716 TI - Correlations among amino acid sites in bHLH protein domains: an information theoretic analysis. AB - An information theoretic approach is used to examine the magnitude and origin of associations among amino acid sites in the basic helix-loop-helix (bHLH) family of transcription factors. Entropy and mutual information values are used to summarize the variability and covariability of amino acids comprising the bHLH domain for 242 sequences. When these quantitative measures are integrated with crystal structure data and summarized using helical wheels, they provide important insights into the evolution of three-dimensional structure in these proteins. We show that amino acid sites in the bHLH domain known to pack against each other have very low entropy values, indicating little residue diversity at these contact sites. Noncontact sites, on the other hand, exhibit significantly larger entropy values, as well as statistically significant levels of mutual information or association among sites. High levels of mutual information indicate significant amounts of intercorrelation among amino acid residues at these various sites. Using computer simulations based on a parametric bootstrap procedure, we are able to partition the observed covariation among various amino acid sites into that arising from phylogenetic (common ancestry) and stochastic causes and those resulting from structural and functional constraints. These results show that a significant amount of the observed covariation among amino acid sites is due to structural/functional constraints, over and above the covariation arising from phylogenetic constraints. These quantitative analyses provide a highly integrated evolutionary picture of the multidimensional dynamics of sequence diversity and protein structure. PMID- 10666717 TI - Polymorphism and divergence in the beta-globin replication origin initiation region. AB - DNA sequence polymorphism and divergence was examined in the vicinity of the human beta-globin gene cluster origin of replication initiation region (IR), a 1.3-kb genomic region located immediately 5' of the adult-expressed beta-globin gene. DNA sequence variation in the replication origin IR and 5 kb of flanking DNA was surveyed in samples drawn from two populations, one African (from the Gambia, West Africa) and the other European (from Oxford, England). In these samples, levels of nucleotide and length polymorphism in the IR were found to be more than two times as high as adjacent non-IR-associated regions (estimates of per-nucleotide heterozygosity were 0.30% and 0.12%, respectively). Most polymorphic positions identified in the origin IR fall within or just adjacent to a 52-bp alternating purine-pyrimidine ((RY)n) sequence repeat. Within- and between-populations divergence is highest in this portion of the IR, and interspecific divergence in the same region, determined by comparison with an orthologous sequence from the chimpanzee, is also pronounced. Higher levels of diversity in this subregion are not, however, primarily attributable to slippage mediated repeat unit changes, as nucleotide substitution contributes disproportionately to allelic heterogeneity. An estimate of helical stability in the sequenced region suggests that the hypervariable (RY)n constitutes the major DNA unwinding element (DUE) of the replication origin IR, the location at which the DNA duplex first unwinds and new strand synthesis begins. These findings suggest that the beta-globin IR experiences a higher underlying rate of neutral mutation than do adjacent genomic regions and that enzyme fidelity associated with the initiation of DNA replication at this origin may be compromised. The significance of these findings for our understanding of eukaryotic replication origin biology is discussed. PMID- 10666718 TI - Weighted neighbor joining: a likelihood-based approach to distance-based phylogeny reconstruction. AB - We introduce a distance-based phylogeny reconstruction method called "weighted neighbor joining," or "Weighbor" for short. As in neighbor joining, two taxa are joined in each iteration; however, the Weighbor criterion for choosing a pair of taxa to join takes into account that errors in distance estimates are exponentially larger for longer distances. The criterion embodies a likelihood function on the distances, which are modeled as correlated Gaussian random variables with different means and variances, computed under a probabilistic model for sequence evolution. The Weighbor criterion consists of two terms, an additivity term and a positivity term, that quantify the implications of joining the pair. The first term evaluates deviations from additivity of the implied external branches, while the second term evaluates confidence that the implied internal branch has a positive branch length. Compared with maximum-likelihood phylogeny reconstruction, Weighbor is much faster, while building trees that are qualitatively and quantitatively similar. Weighbor appears to be relatively immune to the "long branches attract" and "long branch distracts" drawbacks observed with neighbor joining, BIONJ, and parsimony. PMID- 10666719 TI - Did the mitochondrial processing peptidase evolve from a eubacterial regulator of gene expression? PMID- 10666720 TI - A mitochondrial ancestry of the hydrogenosomes of Nyctotherus ovalis. PMID- 10666721 TI - [De novo cancer and adenoma-carcinoma sequence of the colorectum- clinicopathological differences between de novo carcinoma and carcinoma with the sequence]. AB - The adenoma-carcinoma sequence ("The great majority of colorectal carcinomas arises from adenomas") proposed by Morson et al (1972) has been generally accepted world wide. Considering the sequence from the viewpoint of human carcinogenesis in general, however, a few contradictions and mysterious phenomena are found in the development and growth process of the colorectal carcinomas. It is evident that the histogenesis of the adenoma-carcinoma sequence including such contradictions is logically false. Consequently, we should fundamentally reconsider the histogenesis of colorectal carcinoma. The premise for the induction of the histogenesis of colorectal carcinoma is that carcinomas are much more objectively differentiated from adenomas with severe atypia, and that hyperplastic glands with slight atypia are differentiated from adenomas with slight atypia. In order to objectify the histological interpretation of atypical grades, conversion of complicated histological designs into real numbers is required. Two discriminant functions with two variables (N/C ratio and density of tubuli in a unit area) for differentiating carcinoma from adenoma have been determined using computed image processing. Based on the discriminant functions for differentiating objectively between carcinoma, adenoma, and hyperplasia, the histogenesis of colorectal carcinomas has been deduced as follows: 70-80% of colorectal carcinomas arise from the colorectal mucosa (de novo carcinoma) and 20 30% arise from adenoma. Observing the various clinicopathological phenomena in colorectal adenomas and carcinomas from the viewpoint of histogenesis, the contradictions do not enter the logically constructed model. Furthermore, the biological behavior of de novo carcinoma is clinicopathologically different from that of carcinoma with the sequence. PMID- 10666722 TI - [Endoscopic mucosal resection]. AB - Endoscopic mucosal resection (EMR) is a widely used and important technique for the treatment of colorectal neoplasms. The choice must be made between EMR and surgical resection in treating submucosal invading carcinoma. Our study suggests that EMR may be adapted to the treatment of sm1a and sm1b carcinoma without invading vessels. Endoscopic findings suggestive of submucosal invasion are depressed-type tumor, large size, hardness, ulceration on the top of the lesion, etc. We have conducted studies on the surface structure of colorectal neoplasms (pit pattern) and showed that pit patterns are useful in the qualitative and quantitative diagnosis of lesions, when employing magnifying endoscopy. The VN type pit pattern indicates that the lesion is most likely sm-massive cancer. This type of cancer should not be treated by EMR. Although minute colorectal cancer can be treated by EMR. We believe that the importance of endoscopic diagnosis and treatment will increase. PMID- 10666723 TI - [Usefulness of endoscopic polypectomy in early colorectal cancer]. AB - This study was carried out to clarify the usefulness of endoscopic polypectomy in early colorectal cancer. Six hundred and sixty-four patients with polypectomized early colorectal cancer were compared with 89 who underwent surgical resection. Among the polypectomized patients, 361 had pedunculated (Ip)-type, 255 semi pedunculated (Isp)-type, and 48 sessile (Is)-type cancer. The early colorectal cancer was submucosal-invasive cancer (sm cancer) in 16.1% of Ip, 19.2% of I sp, and 22.9% of Is patients. Among the surgically resected patients, sm cancer was present in 41.2% of Ip. 62.1% of I sp. and 58.1% of Is patient in. The percentage of sm cancer and the tumor size surgically resected patients were greater than in polypectomized patients. Local recurrence and distant metastases were investigated after endoscopic polypectomy during follow-up. Two cases (0.4%) of local recurrence were detected after polypectomy among 555 patients with mucosal cancer. Twenty-three (19.8%) of 116 patients with sm cancer underwent surgical resection after polypectomy due to cancer invasion into the vessels and/or extension to the resected margin recognized in polypectomized specimens. Only one case (0.9%) of local recurrence and no distant metastases were detected after polypectomy in 116 patients with sm cancer. Endoscopic polypectomy is very useful as a minimally invasive surgical technique in the protruding type of early colorectal cancer. PMID- 10666724 TI - [Transanal endoscopic microsurgery (TEM)]. AB - Minimally invasive surgery is mandatory for rectal tumors to reduce surgical complications and to improve the quality of life. The conventional transanal procedure is one from of minimally invasive surgery for rectal tumors, but it is still often difficult to reach the middle and upper thirds of the rectum due to anatomical characteristics. Transanal endoscopic microsurgery (TEM) is a technique that allows radical resection of rectal tumors in the distal as well as proximal third of the rectum. Performing radical surgery using this technique requires preoperative estimation of tumor extension both histologically and by X ray images. It should allow provide safe and radical excision of rectal tumors. In our department, patients with rectal tumors are evaluated for risk factors using barium enema, endoscopic ultrasound, punch biopsy, etc. Fifty patients (rectal cancer, 34: cartinoid, 4: adenoma, 12) have undergone radical TEM. The operation was converted to open surgery in 2 cases because of rectal perforation. Radical open surgery was performed after TEM in 1 patient since cancer invasion was revealed by postoperatively upon histological examination. We conclude that TEM is a useful technique for radical resection of rectal tumors and is associated with a low rate of postoperative complications and a high surgical success rate. PMID- 10666725 TI - [Minimally invasive transanal surgery]. AB - A new anal retractor connected to an Octopus retractor holder and automatic suture clamp were used for local excision of rectal tumors to obtain easy access to proximal tumors with an adequate surgical field and to avoid complications. Shortening techniques and invagination techniques were adopted to pull down proximal tumors. This surgical procedure resulted in shorter operating time, less bleeding, earlier oral intake, shorter hospital stay, and fewer complications. PMID- 10666726 TI - [Laparoscopic bowel resection for early colon cancer]. AB - Following the successful introduction of laparoscopic cholecystectomy, many reports confirming the feasibility of using laparoscopy for bowel resection and predicting that it would be advantageous in terms of its minimal invasiveness have been published. In the context of cancer treatment, however, the feasibility of lymphadenectomy, the risk of recurrence, and survival have emerged as major concerns. Even though mucosal cancer (Tis) can be treated by endoscopic resection (ER), when this is not possible open surgery (OS) must be performed. In patients with T1 cancer, tumors showing slight submucosal layer invasion (sm 1) can be treated in the same way as Tis (in cancer) cancers. But 5% to 10% of patients with T1 cancer have massive submucosal layer invasion (sm 2-3) with paracolic lymph node metastasis. At least partial bowel resection with paracolic lymphadenectomy is considered necessary for T1 (sm 2-3) cancers in principle. In summary, laparoscopic local excision of Tis cancers that are endoscopically unresectable and laparoscopically assisted partial resection with paracolic lymphadenectomy for T1 cancers have become accepted because local excision and partial resection with paracolic lymphnedectomy are fairly simple to perform laparoscopically. Therefore as a strategy for the treatment of early colorectal cancer (CRC), minimally invasive laparoscopic bowel resection (LBR) has been positioned between endoscopic resection (ER) and open surgery (OS). While the difficulty of performing radical lymphadenectomy is considered one of the greatest obstacles to the introduction of laparoscopic bowel resection (LBR) for the treatment of advanced colorectal cancer (CRC), early colon cancer is a good indication for laparoscopic bowel resection. PMID- 10666727 TI - [Laparoscopic surgery for early rectal carcinoma]. AB - The indications, techniques, and results of laparoscopic surgery for early rectal carcinoma are described in detail. Laparoscopic surgery is indicated when a mucosal tumor is too large to perform endoscopic or transanal resection or the tumor invades the submucosal layer. When the tumor is located in the Rs or Ra region, surgery can be completed laparoscopically. After dissection of the mesenterium and lymphadenectomy are performed, the anal side of the rectum is divided using EndoGIA II. When the tumor is located in the Rb portion, it is impossible to resect the rectum intracorporeally. In this situation, aperi anal maneuver is essential. The rectal mucosa is circumferentially incised just above the dentate line and the internal anal sphincter is dissected. Dissection is advanced to the intersphincteric space. Dissection between the rectum and the levator ani muscle is completed, and the rectum is pulled through the anus. After the oral side of the rectum is divided, a J-pouch is mase and J-pouch-anal anastomosis is performed. Forty-seven patients with rectal carcinoma were operated upon laparoscopically. Postoperative recovery was better than that after open surgery. Serious intraoperative or postoperative complications have not been encountered in this series. In conclusion, laparoscopic surgery is thought to be the procedure of first choice for early rectal carcinoma. PMID- 10666728 TI - [First successful bilateral living-donor lobar lung transplantation in Japan]. AB - No successful lung transplantation has been reported in Japan until recently when we performed the first successful bilateral living-donor lobar lung transplantation. A 24-year-old woman with primary ciliary dyskinesia began experiencing severe respiratory insufficiency and required mechanical ventilation. On October 28, 1998, she underwent bilateral living-donor lobar transplantation, receiving her sister's lower right lobe and her mother's left lower lobe under cardiopulmonary bypass. The patient was discharged from the hospital 61 days after transplantation. Six months postoperatively, she has returned to normal life and is able to carry out daily activities. She is in good physical condition with a vital capacity of 1.77 L. PMID- 10666729 TI - [Treatment of an iliac artery pseudoaneurysm managed with a stent graft]. AB - Recent developments have extended the indications for endovascular intervention to include endovascular lesions, such as aneurysms and sclerotic arterioocclusive disease. Although conventional treatment of pseudoaneurysms is mainly surgical intervention, the development of stent grafts has made this treatment less invasive than previously. We placed a stent graft in a pseudoaneurysm of the iliac artery with good results, as described in this report. Treatment of a pseudoaneurysm with a stent graft avoids the risk associated with general anesthesia and reduces surgical invasiveness, similar to laparotomy. We expect that this therapeutic mode will be developed further in the future. PMID- 10666730 TI - [Experience of orthotopic liver transplantation from non-heart-beating donors at the University of Pittsburgh Medical Center]. AB - Eight cases of orthotopic liver transplantation (OLT) from non-heart-beating donors (NHBDs) were experienced at the University of Pittsburgh Medical Center. Four cases were from donors whose organs were procured following cardiopulmonary resuscitation (uncontrolled NHBD), and the remaining four hepatic allografts were recovered after sustained cardiopulmonary arrest following extubation in an operating room (controlled NHBD). After OLT from uncontrolled NHBDS, two allografts failed due to preservation injury and hepatic arterial thrombosis, and one showed poor allograft function. In contrast, all four grafts from controlled NHBDs survived and functioned well worked postoperatively. Hepatic allografts from controlled NHBDs is considered to be useful in OLT, especially in Japan where there is a serious brain-dead donor shortage. PMID- 10666731 TI - [Stereotactic neurosurgery for movement disorders]. PMID- 10666732 TI - [Electrophysiological mapping of the trigeminal nerve root during microvascular decompression for trigeminal neuralgia]. AB - A method for intraoperative electrophysiological mapping of the intracranial root of the trigeminal nerve was studied in five patients with trigeminal neuralgia. During surgery, the trigeminal nerve root was stimulated centrally with a bipolar electrode, and antidromic responses were recorded peripherally from three branches of the trigeminal nerve in the face. In all patients, the fibers of the individual subdivisions of the trigeminal nerve root were successfully localized based on the peripheral sites of antidromic response. This neural mapping was used during microvascular decompression in four patients and during a rhizotomy procedure in one patient. As a result of mapping, the fibers of the trigeminal division subserving the pain were clearly confirmed to be compressed by the artery in all four patients who were undergoing microvascular decompression. Likewise, the antidromic responses precisely identified the first division of the trigeminal nerve, which should be preserved to avoid postoperative corneal ulcers in patients undergoing rhizotomy. Based on these findings, it was concluded that this technique enables surgeons to precisely identify which fibers of the trigeminal nerve root should be decompressed or divided during surgery for trigeminal neuralgia. PMID- 10666733 TI - [Epilepsy surgery for focal cortical dysplasia and dysembryoplastic neuroepithelial tumor]. AB - We studied six patients with focal cortical dysplasia (CD) and four patients with dysembryoplastic neuroepithelial tumor (DNT) who had surgical resection for medically intractable epilepsy. In all CD patients, ictal single photon emission computed tomography (SPECT) using 99mTc-ECD revealed hyperperfusion in the regions where magnetic resonance (MR) imaging showed CD abnormalities. Interictal epileptiform activity and ictal seizure onset on electrocorticography using subdural strip or grid electrodes were demonstrated in the CD itself. In contrast, in all DNT patients, interictal SPECT disclosed hypoperfusion in the area of the lesions. Ictal SPECT in one DNT patient disclosed hyperperfusion in the superior area of the region where MR imaging showed cystic abnormalities. Interictal spiking in all DNT patients and ictal seizure onset in two DNT patients were demonstrated not in the lesions themselves but in the distinct zone from the region of the tumor-involved brain. All CD patients who underwent lesionectomy became seizure-free with a mean follow-up period of 33.5 months. All DNT patients who underwent lesionectomy and resection of the epileptogenic cortex became seizure-free or had their seizure significantly reduced a mean follow-up period of 41.5 months. We conclude that CDs have intrinsic epileptogenicity, while DNTs have epileptogenicity not intrinsically but in encompassed cortical surface areas. PMID- 10666734 TI - [An ultrasonic study of the relationship between extracranial carotid atherosclerosis and intracerebral hemorrhage]. AB - We have evaluated the relationship between carotid atherosclerotic change and intracerebral hemorrhage patients. Forty-eight patients with intracerebral hemorrhage treated at our institution were included in this study. Their ages ranged from 38 to 86 years old (average: 61.5 years). There were 28 cases of putaminal hemorrhage and 20 cases of thalamic hemorrhage. Evacuation of hematoma or ventricle drainage was performed in 18 cases within 3 days after the onset of symptoms. The outcome in these cases was that 40 patients survived and 8 patients died. Carotid atherosclerosis was evaluated by B mode-ultrasonography. The severity of carotid atherosclerosis was assessed by using two indicators; incidence of carotid atherosclerosis and maximum percentage diameter of the stenosis. Carotid atherosclerosis on B mode-ultrasonography was detected more frequently in patients with thalamic hemorrhage (84.2%) than in those with putaminal hemorrhage (51.7%). Maximum percentage stenosis of thalamic hemorrhage (17.2 +/- 15.2%) was higher than maximum percentage stenosis of putaminal hemorrhage (6.4 +/- 9.7%). In conclusion, carotid atherosclerosis was an effective indicator of not only ischemical cerebrovascular disease but also of intracerebral hemorrhage. PMID- 10666735 TI - [Spinal dermoid cyst secondary to myelomeningocele repair: a case report]. AB - Dermoid and epidermoid tumors arise from the invagination of epidermal elements into the neural tube during the embryonic period. However, many studies have reported that dermoid and epidermoid tumors occur 5 to 10 years after the first operation on myelomeningocele patients. A seven-year-old male with myelomeningocele which had been repaired in his neonatal period, presented leg pain and deterioration of gait disturbance and urinary incontinence. Neuroradiological examinations revealed a spinal dermoid cyst at the repaired myelomeningocele and tethered spinal cord. We removed the tumor and untethered the tethered spinal cord. Postoperatively, the patient's leg pain, gait disturbance and urinary incontinence improved immediately. Inappropriate surgical treatment of spina bifida may cause a second lesion, which leads to tethered-cord syndrome. We suggest that proper surgical treatment, early checkup and neuroradiological evaluations are very important for spina bifida patients who show signs of neurological deterioration. For the best treatment, neurological, urological and orthopedical follow-up study after the first repair operation is necessary. PMID- 10666736 TI - [A case of rete mirabile with congenital dysplasia of bilateral internal carotid arteries]. AB - We report a case of dysplasia of the congenital bilateral internal carotid arteries with the rete mirabile. The rete mirabile is not usually seen in the course of human growth, but it is a common finding in other mammals. Accordingly, some investigators have thought that the rete mirabile is "developmental shift". Our case has dysplasia of the bilateral internal carotid arteries (one is aplasia and the other is hypoplasia), but the patient had suffered from no ischemic symptom because her brain had been sufficiently fed by each of the rete mirabile. Angiographically, the frequency of the rete mirabile formation is about 1/10,000. There were 20 cases reported until 1997 (including our case). There were 5 cases (27.8%) with ischemic symptoms in spite of internal carotid artery dysplasia, 2 cases (11.1%) with intracerebral hemorrhage, 6 cases (33.3%) with subarachnoid hemorrhage (there were only two cases with aneurysm) and 5 cases without symptoms. We have tried to class the rete mirabile by angiographical findings. One is the M type finding resembling moyamoya vessels in stages 3 & 4 of moyamoya disease, and the other is the N type finding resembling a nidus of arteriovenous malformation. M type occurred in younger patients more often than N type, so M type may be the previous stage of N type. PMID- 10666737 TI - [Successful treatment using detachable coils for traumatic carotid cavernous fistula as a complication of transsphenoidal surgery for a pituitary adenoma: a case report]. AB - We report a case of a patient with traumatic carotid cavernous fistula (CCF) caused by transnasal-transsphenoidal surgery, who was successfully treated using detachable coils. A 47-year-old man was admitted to our hospital because of severe headache. He was confirmed to have a nonfunctioning pituitary adenoma with presellar-type sphenoid sinus. Cerebral angiography initially disclosed no vascular lesions. A transnasal-transsphenoidal adenomectomy was performed. When the anterior wall of the sphenoid sinus was dissected with a chisel, the chisel deeply stuck into the posterolateral part of the sinus. Profuse arterial bleeding was observed through the sphenoid sinus. The bleeding was stopped easily by compression and packing with bone wax. The operation was continued, the sellar floor was opened widely and the tumor was removed subtotally. The medial wall of the cavernous sinus was intact. Histological examination revealed a pituitary adenoma. Immediately after surgery, the patient noticed a bruit. He developed chemosis and abducent palsy on the right side. Cerebral angiography displayed a high-flow CCF, which was attributed to the carotid artery injury caused by the transnasal-transsphenoidal surgery. The CCF disappeared after two-staged embolization using detachable coils, 1st transvenous and 2nd transarterial. Ten months later, cerebral angiography showed persistent occlusion of the fistula, and the patient experienced no tumor recurrence. It is suggested that drilling is a safer procedure than using a chisel for dissection of a sphenoid sinus with incomplete pneumatization. Endovascular treatment using detachable coils proved useful to manage the CCF, an unusual complication of transsphenoidal surgery. PMID- 10666738 TI - [Chronic subdural hematoma associated with malignancy: report of three cases]. AB - Three cases of chronic subdural hematoma (CSH) associated with malignancy are reported. Case 1; A one-year-old girl was referred for vomiting and convulsions. Left CSH was removed, and her symptoms disappeared. Cytological examination of chronic subdural hematoma revealed abnormal white blood cells. A clinical diagnosis of acute monocytic leukemia was made after the laboratory examination. Remission was achieved by chemotherapy, but she died one year after the operation. Case 2; A 72-year-old woman was referred for right hemiparesis and urinary incontinence. Left CSH was irrigated, and her clinical symptoms immediately disappeared. Cytological examination of chronic subdural hematoma revealed abnormal white blood cells. A clinical diagnosis of chronic lymphocytic leukemia was made after the laboratory examination. No treatment was given since there were no clinical symptoms of chronic lymphocytic leukemia. Case 3; A 70 year-old woman who had been affected with early gastric cancer and mammary cancer for the previous two years was admitted to our clinic because of headache, right hemiparesis and consciousness disturbance. Left CSH was irrigated, and her clinical symptoms improved. However, there was a tendency to bleed because disseminated intravascular coagulation had occurred, and CT showed bilateral subdural hematoma. A second irrigation was performed, but her symptoms did not improve. Left acute subdural hematoma, which was removed by craniotomy, occurred three days after the second operation. Pathological examination of the outer membrane of the subdural hematoma revealed invasion of adenocarcinoma. She died three days after the third operation. It is recommended that both the cytological and the histological examinations be performed when possible, since they are simple to perform and very useful in some cases. PMID- 10666739 TI - [A case of spontaneous cervical carotid artery dissection managed by using primary stent placement]. AB - The authors report the case of a 35-year-old male who underwent stenting for spontaneous cervical carotid dissection. He presented with sudden onset of hemicrania and left facial palsy followed by left hemiparesis and dysarthria. On admission, carotid angiography revealed postsinus tapering occlusion of the right internal carotid artery. Initially, he was managed with conservative treatment, but his neurological status deteriorated. Findings of brain CT, MRI and IMP-SPECT suggested hypoperfusion of the right cerebrum. A Palmaz stent, 39 mm in length, was successfully placed over the site of the dissection to restore normal patency through the dissected carotid artery. Following stent implantation, his neurological signs improved gradually but completely. Since the procedure, with oral administration of antiplatelet medication, he has suffered no cerebral ischemic events. Follow-up carotid angiography one year after stent implantation showed good patency of the stented segment. The present case emphasizes the usefulness of stenting for spontaneous cervical carotid dissection. PMID- 10666740 TI - Chemical reduction of phosphate on the primitive earth. AB - If phosphorus played a role in the origin of life, some means of concentrating micromolar levels of phosphate (derived from the calcium phosphate mineral apatite), must first have been available. Here we show that simulated (mini)lightning discharges in model prebiotic atmospheres, including only minimally reducing ones, reduce orthophosphates, including apatite, to produce substantial yields of phosphite. Electrical discharges associated with volcanic eruptions could have provided a particularly suitable environment for this process. Production of relatively soluble and reactive phosphite salts could have supplied a pathway by which the first phosphorus atoms were incorporated into (pre)biological systems. PMID- 10666741 TI - Clay catalysis of oligonucleotide formation: kinetics of the reaction of the 5' phosphorimidazolides of nucleotides with the non-basic heterocycles uracil and hypoxanthine. AB - The montmorillonite clay catalyzed condensation of activated monocleotides to oligomers of RNA is a possible first step in the formation of the proposed RNA world. The rate constants for the condensation of the phosphorimidazolide of adenosine were measured previously and these studies have been extended to the phosphorimidazolides of inosine and uridine in the present work to determine of substitution of neutral heterocycles for the basic adenine ring changes the reaction rate or regioselectivity. The oligomerization reactions of the 5' phosphoromidazolides of uridine (ImpU) and inosine (ImpI) on montmorillonite yield oligo(U)s and oligo(I)s as long as heptamers. The rate constants for oligonucleotide formation were determined by measuring the rates of formation of the oligomers by HPLC. Both the apparent rate constants in the reaction mixture and the rate constants on the clay surface were calculated using the partition coefficients of the oligomers between the aqueous and clay phases. The rate constants for trimer formation are much greater than those dimer synthesis but there was little difference in the rate constants for the formation of trimers and higher oligomers. The overall rates of oligomerization of the phosphorimidazolides of purine and pyrimidine nucleosides in the presence of montmorillonite clay are the same suggesting that RNA formed on the primitive Earth could have contained a variety of heterocyclic bases. The rate constants for oligomerization of pyrimidine nucleotides on the clay surface are significantly higher than those of purine nucleotides since the pyrimidine nucleotides bind less strongly to the clay than do the purine nucleotides. The differences in the binding is probably due to Van der Waals interactions between the purine bases and the clay surface. Differences in the basicity of the heterocyclic ring in the nucleotide have little effect on the oligomerization process. PMID- 10666742 TI - Oligomerization of deoxyguanosine 5'-phosphoro-2-methylimidazolide on a polycytidylate template. AB - The oligomerization of deoxyguanosine 5'-phosphoro-2-methylimidazolide on a polycytidylate template is much less efficient than the oligomerization of the corresponding activated ribonucleotide. Nonetheless oligomers containing up to eight nucleotide residues are detected. The products are 3'-5'-linked oligodeoxyribonucleotides capped at the 5'-terminus with a pyrophosphate-linked monomer. PMID- 10666743 TI - Scanning tunnelling microscopy and molecular modelling of xanthine monolayers self-assembled at the solid-liquid interface: relevance to the origin of life. AB - The development of scanning tunnelling microscopy (STM) has allowed examination of inorganic crystalline surfaces and their interactions with organic adsorbates with unparalleled resolution. As a novel technique in origin of life studies, the application of STM is detailed with particular attention paid to the methods employed in the analysis of organic monolayer structures. STM imaging and molecular modelling of self-assembled monolayers of the purine base, xanthine, formed on the surfaces of graphite and molybdenum disulfide are presented as an example. The putative role of such structures in the origin of life is discussed. PMID- 10666744 TI - Chirality amplification--the accumulation principle revisited. AB - The chirality amplification mechanism proposed by Yamagata in 1966, relying on an Accumulation Principle which involved the parity violating energy difference (1 + epsilon) presumed to be operative at each step in the formation of a homochiral biopolymer, is briefly surveyed historically. The Accumulation Principle is then examined analytically and found to be incapable of producing a unique homochiral polymer in any realistic polymerization process. The extension of the Accumulation Principle to crystallizations which afford enantiomorphic crystals is also scrutinized and found to be misapplied and invalid. PMID- 10666745 TI - Did Viking discover life on Mars? AB - A major argument in the claim that life had been discovered during the Viking mission to Mars is that the results obtained in the Labeled Release (LR) experiment are analogous to those observed with terrestrial microorganisms. This assertion is critically examined and found to be implausible. PMID- 10666746 TI - The subpectoral pulse generator site revisited. PMID- 10666747 TI - Unexpected loss of bipolar pacing with implanted dual chamber pacemakers. AB - Bipolar leads are most commonly used in the current practice of pacemaker therapy. In our study of 124 patients implanted with Guidant/Cardiac Pacemakers (CPI) Vigor dual chamber pacemakers, 5 patients had unexpectedly abrupt increases in bipolar lead impedance and pacing threshold 2 weeks to 18 months postimplantation without changes in sensing function. With the lead configuration reprogrammed to unipolar, the lead impedance and pacing threshold were restored to appropriate ranges. The changes in bipolar lead parameters can be caused by the CPI's "Quick Connect" (QC1) header lead system incorporated in these pacemakers. PMID- 10666748 TI - Tissue temperatures and lesion size during irrigated tip catheter radiofrequency ablation: an in vitro comparison of temperature-controlled irrigated tip ablation, power-controlled irrigated tip ablation, and standard temperature controlled ablation. AB - The limited success rate of radiofrequency catheter ablation in patients with ventricular tachycardias related to structural heart disease may be increased by enlarging the lesion size. Irrigated tip catheter ablation is a new method for enlarging the size of the lesion. It was introduced in the power-controlled mode with high power and high infusion rate, and is associated with an increased risk of crater formation, which is related to high tissue temperatures. The present study explored the tissue temperatures during temperature-controlled irrigated tip ablation, comparing it with standard temperature-controlled ablation and power-controlled irrigated tip ablation. In vitro strips of porcine left ventricular myocardium were ablated. Temperature-controlled irrigated tip ablation at target temperatures 60 degrees C, 70 degrees C, and 80 degrees C with infusion of 1 mL saline/min were compared with standard temperature-controlled ablation at 70 degrees C and power-controlled irrigated tip ablation at 40 W, and infusion of 20 mL/min. Lesion size and tissue temperatures were significantly higher during all modes of irrigated tip ablation compared with standard temperature-controlled ablation (P < 0.05). Lesion volume correlated positively with tissue temperature (r = 0.87). The maximum recorded tissue temperature was always 1 mm from the ablation electrode and was 67 +/- 4 degrees C for standard ablation and 93 +/- 6 degrees C, 99 +/- 6 degrees C, and 115 +/- 13 degrees C for temperature-controlled irrigated tip ablation at 60 degrees C, 70 degrees C, and 80 degrees C, respectively, and 112 +/- 12 degrees C for power-controlled irrigated tip ablation, which for irrigated tip ablation was significantly higher than tip temperature (P < 0.0001). Crater formation only occurred at tissue temperatures > 100 degrees C. We conclude that irrigated tip catheter ablation increases lesion size and tissue temperatures compared with standard ablation in the temperature-controlled mode at the same or higher target temperatures and in the power-controlled mode. Furthermore, tissue temperature and delivered power are the best indicators of lesion volume during temperature-controlled ablation. PMID- 10666749 TI - The first year experience with the dual chamber ICD. AB - In July 1997, a dual chamber pacemaker combined with a tiered therapy implantable cardioverter defibrillator (ICD) first became available in the United States. We report the first-year experience of one center in the United States with this dual chamber ICD. Of a total of 174 ICDs, 95 (55%) were dual chamber devices and 79 (45%) were single chamber. New dual chamber ICD insertions averaged 57.4 +/- 8.9 minutes, though there was a learning curve as the last 30 implants averaged 45.1 +/- 6.1 minutes with a negative slope to the regression line of procedure duration (-0.52, P < 0.05). New single chamber ICD implants were 18.5 minutes quicker (38.9 +/- 7.2 minutes). The most challenging implants were dual chamber upgrades (mean procedure duration 64.9 +/- 15.8 minutes), especially if there was a previously implanted pacemaker and ICD at separate sites. Indications for a new dual chamber device were grouped into classic pacemaker indications (52.5%), which comprised the Class I ACC/AHA guidelines, ICD-specific indications (24.6%), and other (23.0%). In the 34 patients undergoing dual chamber upgrade, the classic and ICD-specific groups were equal (47.0% each). Complications were rare (2.8%), though 3 (8.8%) of 34 undergoing a dual chamber upgrade developed late infections requiring explantation. In its first year, the dual chamber ICD has become a common device at our institution comprising 55% of new implants. As experience grows, we anticipate similar usage at most institutions. PMID- 10666750 TI - Early heart rate variations during head-up tilt table testing as a predictor of outcome of the test. AB - Head-up tilt table testing (HUTT) is a useful tool for the diagnosis of unknown origin of syncope. A setback is its duration. This study tries to establish a specific parameter that, according to the heart rate elevation in the test's initial phase, allows a reliable prediction of its outcome. In a prospective study, every patient being under unknown syncope workup was included. A two-phase 20-minute tilt table test was performed. The initial phase was passive, and the second required pharmacological stimulation with isoproterenol. The basal and 5- and 10-minute heart rate values of the passive phase were measured and compared within the group and against negative tests. During a 1-year period, 115 HUTT were performed: 88 were positive and 27 negative. The negative HUTT patients had an increase in HR of 5.05 (+/- 13.5) beats/min at 5 minutes, and 5.79 (+/- 12.9) beats/min at 10 minutes (P = 0.2). Those with a positive HUTT had an increase of 9.05 (+/- 14.5) beats/min at 5 minutes, and of 10 (+/- 13.4) beats/min at 10 minutes (P < 0.001). There were no significant changes in HR when comparing positive to negative HUTT. There is no specific number that allows predication of outcome early in HUTT. Within the group, variations are important. Only a group tendency can be established, which strongly correlates with the results obtained during the test. PMID- 10666751 TI - Importance of ventricular rate after mode switching during low intensity exercise as assessed by clinical symptoms and ventilatory gas exchange. AB - Automatic mode switching from DDD(R) to DDI(R) or VVI(R) pacing modes has improved dual chamber pacing in patients at high risk for supraventricular tachyarrhythmias. However, little is known about the effect of ventricular pacing rate adaptation after mode switching. We conducted a single-blinded, crossover study in 15 patients (58 +/- 21 years) with a DDD pacemaker who had AV block and normal sinus node function to investigate the influence of pacing rate adaptation to intrinsic heart rate during low intensity exercise. Patients performed two tests (A/B) of low intensity treadmill exercise (0.5 W/kg) in randomized order. They initially walked for 6 minutes while paced in DDD mode. The pacing mode was then switched to VVI with a pacing rate of either 70 beats/min (test A) or matched to the intrinsic heart rate (95 +/- 11 beats/min test B). Respiratory gas exchange variables were determined and patients classified the effort before and after mode switching on a Borg scale from 6 to 20. Percentage changes of respiratory gas exchange measurements were significantly larger (O2 consumption: 8.2 +/- 5.0% vs. -0.6 +/- 7.2%; ventilatory equivalent of CO2 exhalation: 5.3 +/- 4.9% vs. 1.5 +/- 4.3%; respiratory exchange ratio: 7.0 +/- 2.2% vs. 3.5 +/- 3.0%; end-tidal CO2: -5.7 +/- 2.9% vs. -1.8 +/- 2.7%; all P < 0.01) and the increase in subjective assessment of the effort tended to be higher (mean increase on Borg scale: 1.6 +/- 1.9 vs. 1.1 +/- 1.8, P = 0.07) after heart rate unadjusted than after adjusted mode switching. Mode switching from DDD to VVI pacing is better tolerated and gas exchange measurements are less influenced if ventricular pacing rate is adjusted to the level of physical activity. Thus, pacing rate adjustment should be considered as part of automatic mode switch algorithms. PMID- 10666752 TI - Electrophysiological correlates of transmural linear ablation. AB - The purpose of this study is to describe the characteristics of lesions created using radiofrequency (RF) energy delivered through a saline/foam electrode that is designed to simplify ablation of the isthmus between the tricuspid annulus (TA) and the inferior vena cava (IVC). We compared the changes in the electrophysiological parameters produced by the ablation to histological findings. In search of a more practical and effective atrial flutter ablation method, various energy modifications and catheter designs have been tested. It was shown that the efficiency of RF ablation could be improved using an endocardial cooled catheter; resulting in increased lesion size. Thus, we postulate that a similar advantage of the cooled catheter system would allow efficient RF delivery through specially designed long foam electrodes, therefore improving the practicality of TA-IVC isthmus ablation for atrial flutter. The study was performed in two acute and five subacute sheep under general anesthesia and with adequate heparinization. We used a linear ablation catheter system equipped with two 2-cm saline bipole electrode pockets with 1.5-mm separation, each consisting of two 8-mm electrodes with 1-mm spacing, allowing for bipolar pacing and recording. This saline/foam electrode pair were positioned on a support loop. RF energy was applied to the saline electrodes at 50 watts for 90 seconds with a saline flow rate of 0.4 mL/s. Bipolar atrial signal amplitude and pacing thresholds were measured before and after ablation. If necessary, the catheter was pulled back and additional ablation was applied to any viable tissue. Transisthmus ablations were created with a single catheter positioning in five sheep using both saline electrodes in four and one electrode in the other. Pullback and additional ablation to one saline electrode was required in two sheep; in one after RF was delivered to only one electrode. After ablation, atrial signal amplitude was reduced by an average of 76% (range 51%-92%) and its pacing threshold was increased by an average of 617% (range 150%-400%). Transmural lesions were found in all sheep, measuring 8-20 mm in length, 4-10 mm in width, and 1.5-2.0 mm in depth. No charring, coagulum, or remote structural damage was found in any preparation. Continuous transmural TA-IVC isthmus lesions could be produced with stationary RF linear ablation using a saline/foam electrode catheter system. This system allowed for assessment of electrophysiological parameters that correlated with complete necrosis. PMID- 10666753 TI - Long-term reproducibility of ventricular tachycardia induction with electrophysiological testing in patients with coronary heart disease and depressed left ventricular ejection fraction. AB - The Multicenter Automatic Defibrillator Implantation Trial (MADIT) has recently confirmed the role of programmed ventricular stimulation (PVS) to identify the high risk patients of sudden death after myocardial infarction and to prevent this risk. The purpose of this study was to evaluate the long-term reproducibility of PVS in these patients. Thirty patients with coronary heart disease without spontaneous documented sustained ventricular tachycardia (VT) underwent two programmed stimulations in the absence of antiarrhythmic drug treatment between 2 and 6 years (mean 4 years). No patient had a myocardial infarction or intervening cardiac surgery during this period. The protocol of study was similar using up to three extrastimuli in two sites of the right ventricle, delivered in sinus rhythm and driven rhythm (600 ms, 400 ms, respectively). On the first PVS, 17 patients had inducible sustained VT (group I). Thirteen patients did not have inducible VT (group II). On the second PVS all group I patients but one had inducible VT, but the cycle length was significantly modified in 11. In group II, five patients had inducible VT and in the other patients the PVS remained negative. In conclusion, in patients with coronary heart disease, but without documented VT, the long-term reproducibility of PVS was excellent in those with inducible VT (94%); the patients remain at risk of VT and a prophylactic implantable cardioverter defibrillator could be considered. In patients with initially negative study, reproducibility of PVS was lower (61.5%), probably because of the progressive remodeling after myocardial infarction. Therefore, the occurrence of new symptoms in patients with previously negative study requires a second programmed ventricular stimulation. PMID- 10666754 TI - The recording of monophasic action potentials with fractal-coated iridium electrodes in humans. AB - The present study was designed to evaluate the feasibility of the recording of monophasic action potentials (MAP) with fractal-coated iridium electrodes in a clinical setting. In 18 patients who underwent an electrophysiological study for various arrhythmias, we performed MAP recordings with both 1.3-mm2 and 6-mm2 tip surface area fractal-coated iridium and standard silver--silver chloride (Ag/AgCl) electrodes in the high right atrium and two ventricular positions. Amplitude and MAP duration at 90%, 50%, and 25% of repolarization were calculated during steady-state pacing at 600, 500, and 400 ms cycle lengths with extrastimuli application. Morphology comparisons of MAP signals recorded with both types of electrodes were performed by regression analysis using 5% of the repolarization segments of the MAP trajectory. Differences between MAP duration at 90%, 50%, and 25% of repolarization recorded with fractal-coated and Ag/AgCl electrodes were statistically insignificant. Amplitude values recorded with 6-mm2 tip electrodes were significantly smaller than those recorded with Ag/AgCl electrodes for all comparisons. During steady-state pacing, the correlation coefficients between Ag/AgCl and fractal-coated 1.3-mm2 and 6-mm2 tip electrodes were within the range of 0.93-0.999 and 0.87-0.999, respectively. The correlation of MAP amplitude and duration at 90%, 50%, and 25% of repolarization following the extrastimulus S2, recorded with both types of electrodes, was significantly weaker for right atrial recordings (r value range 0.78-0.92) as compared to ventricular recordings (r value range 0.92-0.99). The MAP sensing features of fractal-coated iridium and Ag/AgCl electrodes are comparable. The best results for recording of MAPs with fractal-coated electrodes can be achieved with small surface area tip electrodes. PMID- 10666755 TI - Impact of transisthmus linear ablation of typical atrial flutter on coronary sinus activation time. AB - Complete or incomplete bidirectional isthmus conduction block after linear ablation of atrial flutter is difficult to interpret without detailed multiple electrodes mapping along the tricuspid annulus and the low right atrial isthmus area. The influence of isthmus block on the intraatrial septal and coronary sinus activation has not been assessed by endocardial mapping. This study was designed to analyze the intraartial and interatrial activation times in a retrospective fashion to investigate (1) whether isthmus conduction block can change the coronary sinus activation sequence during low lateral right atrial pacing, and (2) the correlation between change of coronary sinus activation time and isthmus conduction block. Sixty-five consecutive patients (mean age, 57 +/- 18 years) with clinically documented typical atrial flutter were studied. A 20-pole "Halo" catheter was placed around the tricuspid annulus including the entire low right atrial isthmus to verify complete bidirectional isthmus block. Activation time from ostium to distal coronary sinus (OCS-->DCS), and interatrial septum and isthmus activation times during right atrial pacing were analyzed and compared before and after incomplete or complete isthmus block. Complete bidirectional isthmus block was achieved in 50 (77%) patients. During low lateral right atrial pacing, linear ablation at low right atrial isthmus results in a significant delay of activation in all coronary sinus recording sites with greater extent at the ostium area without influence on interatrial septum activation in complete and incomplete isthmus conduction block. The difference of the OCS-->DCS interval before and after ablation, delta (OCS-->DCS), was well correlated with results of isthmus conduction block and significantly longer in patients with complete than those with incomplete isthmus block (34 +/- 11 vs 11 +/- 8 ms, P < 0.001), thereby allowing a value of 20 ms as a discriminative parameter to differentiate incomplete (< 20 ms) from complete (> or = 20 ms) isthmus counterclockwise conduction block with a sensitivity of 96% and a specificity of 88%. In conclusion, creation of a line of block at the inferior vena cava-tricuspid annulus isthmus could change coronary sinus activation sequence during low lateral right atrial pacing in sinus rhythm. The change of coronary sinus activation time after linear ablation, delta (OCS-->DCS), was well correlated with isthmus conduction block by using a value > or = 20 ms to discern complete counterclockwise isthmus block. PMID- 10666756 TI - Preserving normal ventricular activation versus atrioventricular delay optimization during pacing: the role of intrinsic atrioventricular conduction and pacing rate. AB - The purpose of the study was to compare the effects of DDD pacing with optimal AV delay and AAI pacing on the systolic and diastolic performance at rest in patients with prolonged intrinsic AV conduction (first-degree AV block). We studied 17 patients (8 men, aged 69 +/- 9 years) with dual chamber pacemakers implanted for sick sinus syndrome in 15 patients and paroxysmal high degree AV block in 2 patients. Aortic flow and mitral flow were evaluated using Doppler echocardiography. Study protocol included the determination of the optimal AV delay in the DDD mode and comparison between AAI and DDD with optimal AV delay for pacing rate 70/min and 90/min. Stimulus-R interval during AAI (ARI) was 282 +/- 68 ms for rate 70/min and 330 +/- 98 ms for rate 90/min (P < 0.01). The optimal AV delay was 159 +/- 22 ms. AV delay optimization resulted in an increase of an aortic flow time velocity integral (AFTVI) of 16% +/- 9%. At rate 70/min the patients with ARI < or = 270 ms had higher AFTVI in AAI than in DDD (0.214 +/ 0.05 m vs 0.196 +/- 0.05 m, P < 0.01), while the patients with ARI > 270 ms demonstrated greater AFTVI under DDD compared to AAI (0.192 +/- 0.03 m vs 0.166 +/- 0.02 m, P < 0.01). At rate 90/min AFTVI was higher during DDD than AAI (0.183 +/- 0.03 m vs 0.162 +/- 0.03 m, P < 0.01). Mitral flow time velocity integral (MFTVI) at rate 70/min was higher in DDD than in AAI (0.189 +/- 0.05 m vs 0.173 +/- 0.05 m, P < 0.01), while at rate 90/min the difference was not significant in favor of DDD (0.149 +/- 0.05 m vs 0.158 +/- 0.04 m). The results suggest that in patients with first-degree AV block the relative impact of DDD and AAI pacing modes on the systolic performance depends on the intrinsic AV conduction time and on pacing rate. PMID- 10666757 TI - The comparative effects of drive and test stimulus intensity on myocardial excitability and vulnerability. AB - The number and intensity of stimuli that set basic cycle length in cardiac electrophysiological studies can influence the electrical properties assessed by extrastimuli. The relative contribution of drive (S1) and test (S2) stimulus intensity in defining myocardial excitability and vulnerability has not been reported. The purpose of this investigation was to assess this interaction and to determine whether atrial and ventricular findings differed. The effects of S1 and S2 intensity on atrial and ventricular stimulus-intensity-refractory-period curves were determined in open-chest dogs: comparisons were made between curves with S1 intensity varied between diastolic threshold (DT) and 10 mA and S2 intensity maintained at DT and those with S1 intensity maintained at DT and S2 intensity varied between DT and 10 mA. S1-S1 was held constant and S1-S2 varied. The effects of different stimulation sites, cycle length, number of stimulations, and neural blockade were assessed. S1 intensity amplification shifted atrial stimulus-intensity-refractory period curves in the direction of increased excitability and vulnerability; the changes were more pronounced than those obtained by modulating S2 intensity. The changes produced by increasing S1 intensity were evident at different cycle lengths and were enhanced by an increased number of stimulations, but were not evident when S1 and S2 were delivered at different atrial sites. Although beta-blockade attenuated the effects of increasing S1 intensity somewhat, the addition of cholinergic blockade virtually abolished it. Ventricular refractoriness was also changed by modulation of S1 intensity, but the changes were less striking. In the atrium, modulation of S1 intensity has greater effects of stimulus-intensity-refractory-period relations than modulation of S2 intensity; in the ventricule, the converse is true. PMID- 10666758 TI - Electrophysiologist-implanted transvenous cardioverter defibrillators using local versus general anesthesia. AB - With the advent of smaller biphasic transvenous implantable cardioverter defibrillators (ICDs) and the experience gained over the years, it is now feasible for electrophysiologists to implant them safely in the abdominal or pectoral area without surgical assistance. Throughout the years, general anesthesia has been used as the standard technique of anesthesia for these procedures. However, use of local anesthesia combined with deep sedation only for defibrillation threshold (DFT) testing might further facilitate and simplify these procedures. The purpose of this study was to test the feasibility of using local anesthesia and compare it with the standard technique of general anesthesia, during implantation of transvenous ICDs performed by an electrophysiologist in the electrophysiology laboratory. For over 4 years in the electrophysiology laboratory, we have implanted transvenous ICDs in 90 consecutive patients (84 men and 6 women, aged 58 +/- 15 years). Early on, general anesthesia was used (n = 40, group I), but in recent series (n = 50, group II) local anesthesia was combined with deep sedation for DFT testing. Patients had coronary (n = 58) or valvular (n = 4) disease, cardiomyopathy (n = 25) or no organic disease (n = 3), a mean left ventricular ejection fraction of 35%, and presented with ventricular tachycardia (n = 72) or fibrillation (n = 16), or syncope (n = 2). One-lead ICD systems were used in 74 patients, two-lead systems in 10 patients, and an AVICD in 6 patients. ICDs were implanted in abdominal (n = 17, all in group I) or more recently in pectoral (n = 73) pockets. The DFT averaged 9.7 +/- 3.6 J and 10.2 +/- 3.6 J in the two groups, respectively (P = NS) and there were no differences in pace-sense thresholds. The total procedural duration was shorter (2.1 +/- 0.5 hours) in group II (all pectoral implants) compared with 23 pectoral implants of group I (2.9 +/- 0.5 hours) (P < 0.0001). Biphasic devices were used in all patients and active shell devices in 67 patients; no patient needed a subcutaneous patch. There were six complications (7%), four in group I and two in group II: one pulmonary edema and one respiratory insufficiency that delayed extubation for 3 hours in a patient with prior lung resection, both probably related to general anesthesia, one lead insulation break that required reoperation on day 3, two pocket hematomas, and one pneumothorax. There was one postoperative arrhythmic death at 48 hours in group I. No infections occurred. Patients were discharged at a mean time of 3 days. All devices functioned well at predischarge testing. Thus, it is feasible to use local anesthesia for current ICD implants to expedite the procedure and avoid general anesthesia related cost and possible complications. PMID- 10666759 TI - The genetics of cardiac arrhythmias. PMID- 10666760 TI - Inappropriate antitachycardia pacing: a dangerous component failure or pseudotherapy? PMID- 10666761 TI - Short-, mid-, and long-term reproducibility of the atrial signal-averaged electrocardiogram in healthy subjects: comparison with the conventional ventricular signal-averaged electrocardiogram. AB - Although atrial signal-averaged electrocardiogram (SAECG) has been proposed for noninvasive identification of patients with atrial tachyarrhythmias, the substantial variability of the measurement limits the clinical value. The aim of the study was to assess the short- to long-term reproducibility of atrial SAECG and to compare it to that of the conventional ventricular SAECG in 51 healthy volunteers (30 men; age 32 +/- 8 years). In each subject, SAECG recordings were obtained using MAC-VU electrocardiograph and HiRES and PHiRES software (Marquette Medical Systems) and repeated after 5 minutes, 1 day, 1 week, and 1 month. Automatically detected onset and offset of the filtered QRS complex and P wave were subsequently corrected by two independent observers, and the averaged values were used for the analysis. Conventional ventricular SAECG parameters: filtered QRS duration (QRStot), low amplitude signal duration, and root mean square voltage (RMS) of the terminal 40 ms of QRS, and 5 atrial parameters: filtered P wave duration (Ptot), RMS of the terminal 40, 30, 20 ms, and of the entire P wave were obtained. Relative errors of different pairs of measures were used to assess the intrasubject reproducibility. QRStot and Ptot were the most reproducible parameters. The relative errors after 5 minutes, 1 day, 1 week, and 1 month were 0.8% to 2.4% for QRStot, and 1.3% to 4.2% for Ptot. For RMS voltages, the relative errors exceeded 15% in short-term and 20% in long-term recordings. Although Ptot was statistically less reproducible than QRStot, the reproducibility of the former was good and comparable to that of the QRStot. The reproducibility of the voltage parameters was significantly poorer than that of the duration parameters. The study showed a satisfactory short- and long-term reproducibility of Ptot in the atrial SAECG in healthy subjects. However, low reproducibility of the voltage parameters should be considered in clinical applications. PMID- 10666762 TI - The physician's office: now accrediting the environment of care.... PMID- 10666763 TI - Cisapride and torsades de pointes in a pacemaker patient. PMID- 10666764 TI - Radiofrequency ablation of a posteroseptal atrioventricular accessory pathway in a left-sided tricuspid ring with Ebsteinlike anomaly in a patient with congenitally corrected transposition of the great arteries. AB - Radiofrequency ablation successfully eliminated a posteroseptal accessory pathway in a left-sided tricuspid ring with Ebsteinlike anomaly in a patient with a congenitally corrected transposition of the great arteries. PMID- 10666765 TI - Successful treatment of severe orthostatic hypotension with cardiac tachypacing in dual chamber pacemakers. AB - Orthostatic hypotension is an evolving and disabling disease usually observed in elderly patients with dramatic consequences on morbidity, mortality, and impairing the quality of life. We studied the effects of the pacing rate and AV interval on the blood pressure drop in the upright position in two patients with previously implanted pacemakers for sinus node dysfunction. Although the AV interval did not affect the blood pressure drop in the upright position, tachypacing at 100 paces/min improved it dramatically and prevented syncope. Cardiac tachypacing is a useful therapeutic option in severe refractory orthostatic hypotensive patients, especially those with chronotropic incompetence. PMID- 10666766 TI - Adenosine induced atrial fibrillation precipitating polymorphic ventricular tachycardia. AB - An 86-year-old female developed supraventricular tachycardia 36 hours after a myocardial infarction (MI). She developed atrial fibrillation and polymorphic ventricular tachycardia (PVT) following administration of 12 mg of adenosine. The PVT caused hemodynamic instability with no response to cardioversion, but termination with procainamide. The heart is vulnerable to hemodynamically unstable, possibly lethal, PVT early after MI under some circumstances. This vulnerability may be exposed following administration of adenosine. Extra caution is warranted when using adenosine in the post-MI period. PMID- 10666767 TI - Dual chamber pacemaker implantation via a double superior vena cava. AB - We present the chest X ray of a woman with sinoatrial node disease. She has had a dual chamber pacemaker implanted with each lead coursing through a separate superior vena cava. PMID- 10666768 TI - Pacing approaches to the patient with a univentricular heart and the factors associated with choice of pacing site. PMID- 10666769 TI - Natural and induced regulation of Th1/Th2 balance. AB - Because Th1/Th2 balance is perturbed during immunological disease, the design of strategies aiming at its rectification has become a priority. The alteration of the balance in pregnancy so as to promote survival of the fetal allograft lends credibility to this aim. Attenuation of the activation signal delivered through the T cell receptor (TCR) represents a promising approach. It is supported by the high level of polymorphism in the MHC class II promoter, which regulates the natural TCR signal and thus modulates Th1/Th2 differentiation. Further support comes from the Th2 shift that occurs in JNK knockout mice, and with kinase inhibitors and anti-CD4 monoclonal antibodies applied in vitro. The approach has implications for nasal tolerance and inhibition of IL-12 production. The further range of options for Th1/Th2 modulation, which are presented throughout this issue of the journal, are here summarised and evaluated. PMID- 10666770 TI - Molecular mechanisms in T helper phenotype development. PMID- 10666771 TI - Educating T cells: early events in the differentiation and commitment of cytokine producing CD4+ and CD8+ T cells. AB - T lymphocytes acquire the ability to synthesize cytokines during their primary response to antigen, often giving rise to effector populations with a polarized type 1 or type 2 cytokine profile. However, polarization is not a simple choice between two differentiation pathways. This article reviews the evidence, particularly from single-cell and clonal studies, that polarization is the outcome of a series of stochastic events whose probabilities are determined in part by genetic background and in part by extracellular signals received during activation and clonal expansion. The data suggest that these extracellular signals independently and differentially regulate the probability of expression of each cytokine gene, for example by their effects on clonal expansion and chromatin remodeling, CpG demethylation and transcriptional activation of cytokine genes. Polarization is, therefore, achieved at the population level by altering frequencies of expression among cells with many different expression patterns, rather than by selective differentiation of a discrete subset. Type 1 and type 2 populations progressively lose responsiveness to counter-polarizing stimuli. While the molecular basis of this process is not yet known, the observed persistence of cells with flexible cytokine profiles in some polarized populations suggests that loss of flexibility may also be a probabilistic event. PMID- 10666772 TI - The critical role of IL-12 and the IL-12R beta 2 subunit in the generation of pathogenic autoreactive Th1 cells. AB - Experimental Allergic Encephalomyelitis (EAE) is a demyelinating disease of the central nervous system which is an animal model for the human autoimmune disease, multiple sclerosis. EAE is mediated by CD4+ T cells and the T cells responsible for disease induction produce Th1 cytokines. IL-12 produced by monocytes and dendritic cells is the most critical factor which influences the development and differentiation of pathogenic autoreactive Th1 cells. Here, we review our recent studies on the critical contributions of IL-12 and the IL-12R beta 2 subunit to the generation of autoreactive effector cells which mediate EAE. In addition, we discuss the potential contribution of IL-18 to the upregulation of the IL-12/IL 12R beta 2 pathway and the contribution of the suppressor cytokines, IL-4 and IL 10, in downregulating this pathway. Collectively, our studies demonstrate that the IL-12/IL-12R beta 2 pathway is a critical intermediary in the process of Th1 differentiation which can be both positively or negatively regulated. This pathway remains an attractive immunotherapeutic target for blockade of function with inhibitory reagents or downregulation by Th2 cytokines. PMID- 10666773 TI - Th1/Th2 subsets: distinct differences in homing and chemokine receptor expression? AB - The functional specialization of T effector cells according to cytokine secretion patterns has been recognized as an important parameter shaping local immune responses. Here we discuss evidence that T cell subsets might also develop distinctive properties related to homing and trafficking into inflamed sites. First, ligands for the inflammation-induced endothelial selectins were found to be induced by IL-12, and hence selectively expressed on Th1 cells generated in vitro. However, their expression on effector cells occurring in vivo is less well correlated with the Th subset. Second, a variety of receptors for and responses towards chemokines have been found to be differentially associated with Th subsets. Notably CCR5 and, to a lesser degree CXCR3 were preferentially found on Th1 cells, CCR4, CCR8 and, more controversial, CCR3 and CXCR4 on Th2 cells. Although many points, such as stability of the phenotype versus dependency on inducing cytokines and activation stages remain to be clarified, it appears that this field provides new insights into the regulation of locally balanced activities of Th subsets and might constitute a promising field for the development of new immunosuppressive drugs. PMID- 10666774 TI - Control of immune pathology by IL-10-secreting regulatory T cells. PMID- 10666775 TI - Th1/Th2 balance in atopy. PMID- 10666776 TI - Th1/Th2 balance in infection. AB - Cytokines produced by T helper (Th) cells are of critical importance for the outcome of many infectious diseases. Producing the "right" set of cytokines in response to an infectious agent can be a matter of life or death. The Th1/Th2 dichotomy, although an oversimplification has proven useful in the analysis of immune responses to infections. In some infectious diseases, most notably leishmaniasis or infections with gastrointestinal helminths, one Th subset is indispensable for clearing the infection, whereas the opposite Th subset is detrimental. More frequently, both Th1 and Th2 responses are required at different time points to effectively eradicate an infectious agent. The granuloma responses to either Mycobacterium tuberculosis or Schistosoma mansoni provide illustrative examples and are discussed in this review. There is accumulating evidence for frequent coexpression of Th1 and Th2 cytokines during the in vivo immune response to infections. The mechanisms by which infectious agents modulate Th1/Th2 phenotype development are summarized here. Finally, we review here the current evidence for cytokine imbalances induced by infections as pathogenic or protective factors in autoimmunity and allergy. PMID- 10666777 TI - Th1/Th2 balance in cancer, transplantation and pregnancy. PMID- 10666778 TI - Type 1 and type 2 immune responses in children: their relevance in juvenile arthritis. PMID- 10666779 TI - Centenarians in the United States, 1990 and beyond. AB - Persons who live to be 100 years-old and older constitute a fascinating cohort. The number of these centenarians continues to grow fairly rapidly in the United States, with less rapid growth evident in various other developed countries. Characteristics of this cohort are studied with great interest in the hope of discovering secrets of longevity and better understanding the aging process. Using data from the 1990 Census of Population, this report examines the number of centenarians enumerated in 1990 and reports projections for the future. In 1990, 37,306 persons were classified as centenarians but later evaluation determined that this number was excessive. The Census Bureau now indicates the number approximates 28,000 U.S centenarians for 1990. The 1990 data show that centenarians were more likely to be women that men and to have lower educational levels than those in younger cohorts. Most centenarians are widowed and around one-half live in nursing homes. Looking to the future, the number of centenarians is projected to increase dramatically and there will be greater racial and ethnic diversity among those surviving to age 100 and above in the decades to come. PMID- 10666780 TI - Cardiac catheterizations: average inpatient charges, 1998. AB - In 1998 the average change for an impatient cardiac catheterization (CC)was $12,450 among 13,922 group health insured over age 30. Among the 29 states in which at least 150,CCs were performed, the average total charge ranged from $24,000 in California, which was 93 percent above the U.S. norm, to just over one third of this total in Iowa ($8,810). The second highest average charge was reported in Texas ($20,140, 62 percent above the norm) and the second lowest was in Maryland ($11,420, 8 percent below). On average, the hospital proportion of the total CC charges accounted for 80 percent but ranged from 86 percent in California to 71 percent in Maryland. Physician fees averaged $2,450 across the country and ranged from $3,830 in Texas (56 percent above in average) to $2,140 in Iowa (13 percent below the norm). Length to stay averaged 3.2 days, with patients in Iowa remaining in the hospital for 5.6 days and those in Washington 2.9 days. Per diem costs averaged $3,850 and were the highest in California, $6,470 (68 percent above the average) and $1,570 in Iowa (59 percent below). PMID- 10666781 TI - Confidence and confusion: the health care system in the United States. AB - Results from the latest "Health Confidence Survey" show that Americans generally appear to be happy with their health care system experiences. Satisfaction with the care received over the past two years and with current health insurance plans remains high and virtually unchanged since the 1998 survey. Close to nine out of 10 respondents were satisfied with their health plan in 1999 as well as the quality of care received. Confidence in many aspects of the health care system is high--94 percent of Americans report being confident that their pharmacist will fill their prescription correctly, 90 percent are confident they will be able to see a health care specialist, as needed, and 89 percent have confidence in the quality of care that hospitals deliver. Confidence is not as high, however, for belief in the confidentiality of medical records or that a fair review will be be received if health insurance coverage is denied. While Americans are highly confident in most aspects of the health care system today, they continue to be less so for the next 10 years and even less so once they become Medicare eligible. Confidence in the future of health care may be low because Americans appear to be uncertain about managed care and what constitutes a managed care plan. While 71 percent of those enrolled in a managed care plan label it as a PPO or HMO, only 21 percent reported that they are currently enrolled ina managed care plan. The fact that so many Americans do not know that their health plan is a managed care plan lends support to our contention that these plans, benefits managers, employers, policymakers and the media have failed to educate enrollees and the general public about the features and potential advantages of such plans. PMID- 10666782 TI - KIDS COUNT: identifying and helping America's most vulnerable. AB - Using data from the 1999 KIDS COUNT Data Book, compiled and published by The Annie E. Casey Foundation, the most recent state figures (1996) are compared with corresponding data from 1985 to assess the trends in child well-being in each state during the decade. The 10 key indicators used to rank states are taken fro federal government statistical agencies and reflect the best data available for each of the measures. Between 1985 and 1996, child well-being improved across the country in seven of the indicators (infant mortality rate; child death rate; rate of teen death by accidents, homicide and suicide; percent of teens who are high school dropouts; percent of teens not attending school and not working; percent of children living with parents who do not have full-time, year-round employment; and percent of children living in poverty). The three areas of children's well being that worsened during the time period were; the percent of low birth-weight babies; teen birth rate; and percent of families with children headed by a single parent. While many of America's children have experienced improvements in their well-being, there is a large number of children who have been left behind. The focus of the 1999 KIDS COUNT Date Book is on the 9.2 million children who have been identified as "high risk." These children have four or more family disadvantages that put them at greater risk of experiencing poor outcomes. These kids are not benefiting from the continuing economic boom in the 1990s. The District of Columbia, Louisiana and Mississippi had the largest share of high risk kids, ranging from 39 to 21 percent of their respective populations, and Utah the lowest, with just 5 percent. Long-term family-centered strategies that are multi-dimensional and community-wide are recommended to help promote family change. PMID- 10666783 TI - Computer use among the U.S. school-aged population, 1989, 1993 and 1997. PMID- 10666784 TI - [Serosurveillance of notifiable veterinary diseases in wild boar in the Netherlands]. AB - During the hunting season 1996-1999, blood samples were collected from wild boar shot in The Netherlands. Sera were screened for presence of antibodies against classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), Aujeszky's disease virus (ADV), and Trichinella spiralis. The results indicate that CSFV, SVDV, and ADV are uncommon in the wild boar population. Therefore, it seems that CSFV, SVDV, and ADV infection in the wild boar population is not an important reservoir in The Netherlands. ADV and CSFV infections are endemic in the wild boar population in Germany. Since contact between the German and Dutch wild boar populations can not be excluded, continuation of the sero-surveillance system seems appropriate. In the decade before 1998, the wild boar population in The Netherlands seemed to be free of T. spiralis. Whether the finding, in the hunting season of 1998-1999, of a few wild boar with antibodies against T. spiralis is an artefact or not, should be investigated in further research. PMID- 10666785 TI - [Operative removal of a bladder stone by urethro-sphincterotomy in a pony mare]. AB - A case of a cystic calculus in a pony mare is described. In this case an urethro sphincterostomy accomplished removal of a 362 g calculus with dimensions of 9 x 7 cm without any complications after surgery. PMID- 10666786 TI - [Slappendel]. PMID- 10666787 TI - [Discussion of regular/alternative medicine from all times]. PMID- 10666788 TI - [Heck cattle in the Oostvaardersplassen: risk for infectious diseases or not?]. AB - This article discusses risk analysis of infectious diseases in a cattle population. Heck cattle living in nature reserve 'De Oostvaardersplassen' were studied as an example of the risk analysis approach. Twenty-five adult cattle were tested for every infectious disease agent that the risk analysis indicated might be prevalent. All sampled cattle tested positive for Bovine Herpesvirus 1, whereas the prevalence of antibodies against other infectious disease agents was below the level that was assumed to be the threshold for spread of the disease to cattle in the surrounding areas. Risk management of infectious diseases was expressed in a so-called 'low-risk profile'. The risk of introduction or spread of infectious diseases was estimated to be very low as long as the population was kept strictly enclosed. PMID- 10666789 TI - [Refusing immunocastrated piglets]. PMID- 10666790 TI - [Transfusion medicine: from myth to reality]. PMID- 10666791 TI - [Transfusion medicine: debate, field of application, the stakes]. AB - Transfusion medicine has aroused much controversy as to the definition of its field of application, as the Etablissement Francais du Sang (EFS) is being set up. One argument put forward by supporters of transfusion medicine in hospital- including nominal attribution and clinical immunohematology--is the possible influence of blood component producers on the decisions of prescribers. The EFS considers transfusion medicine as the rationale underlying a therapeutic chain applied to blood products and possibly to cell therapy products, within a coherent structure. Any disruption in the chain would result in reduced visibility at each stage of the chain, diluted responsibilities, and less efficient communication between blood collection and actual needs. It is, however, extremely important to clearly distinguish between the activities related to the production of biological products for therapeutic use from the activities of evaluation, in the setting of clinical protocols, which are the clinicians' responsibility. The EFS views the production of blood components as one of the various aspects of its missions as a transfusion medicine operator. The involvement of blood transfusion centers in the development of biotechnologies is a historical reality which justifies EFS's ambition to integrate cell therapy activities within its field of competence. The EFS, with its 18 blood transfusion centers, will have to play a part in health care, research and teaching networks, at the regional or inter-regional level. Irrespective of the progress achieved in substitution products and the evolution to be expected in transfusion-related jobs, transfusion as a discipline must maintain its coherence to be able to better assume its responsibilities. PMID- 10666792 TI - [Scientific bases of education in transfusion medicine]. AB - Transfusion medicine is a medical speciality which deals with all aspects of blood transfusion. The recent evolution of this speciality was marked by an increased collaboration with clinical medicine, the development of alternatives to allotransfusion (autotransfusion, growth factors, molecular genetics), and the technical progress in laboratory techniques. The development of basic science boosted the introduction of new techniques in the detection of virus and blood cell typing. The analysis of clinical trials allowed a better definition of guidelines for the use of blood products. The blood transfusion teaching program is limited during medical training. Specialists in transfusion medicine have to complete a post-graduate course to become certified. Continuous medical training is organised to allow the specialists to keep their accreditation. Transfusion medicine is becoming more and more complex. This complexity should not lead to breaking down transfusion medicine into several activities, but on the contrary to uniting this medical speciality, within which the different actors shall remain open-minded and able to use the same 'language'. PMID- 10666793 TI - Vascular biosynthesis of nitric oxide: effect on hemostasis and fibrinolysis. AB - Nitric oxide (NO) is a multifunctional effector molecule that plays a central role in the regulation of vascular homeostasis. NO is synthesized from L-arginine by a family of enzymes called NO synthases. The principal source of NO in the vascular system of healthy mammals is the constitutively expressed NO synthase in endothelial cells. The basal endothelial formation of NO can be increased by receptor-dependent agonists (i.e., bradykinin) in a calcium-calmodulin-dependent manner, and also by physical forces (i.e., shear stress), predominantly without changes in the intracellular concentration of free calcium. Nitric oxide can diffuse toward the blood vessel wall where the major target is the smooth muscle cell. NO regulates vascular tone, and the free radical is also a potent inhibitor of smooth muscle cell proliferation, migration and synthesis of extracellular matrix proteins. NO can also diffuse toward the lumen of the blood vessel where it helps maintain blood fluidity. NO inhibits platelets' and leucocytes' adhesion to endothelial cells. In addition, NO inhibits platelet aggregation and facilitates the dissolution of small platelet aggregates. However, the regulatory action of NO on blood cells is most likely limited to the luminal surface of endothelial cells since NO is rapidly scavenged by hemoglobin in erythrocytes and inactivated by oxygen-derived radicals such as superoxide anions. NO can also affect the fibrinolytic activity by regulating the release of tissue-type plasminogen activator and plasminogen activator inhibitor-1. The crucial role of vascular NO in the control of blood fluidity has been demonstrated by the regulation of the bleeding time in humans. PMID- 10666794 TI - [Developments in cell therapy in the year 2000]. AB - For the past thirty years, hematology has switched from the concept of bone marrow transplantation to the concept of hematopoietic stem cell (HSC) transplantation, from allograft to autograft, from non-manipulated graft to hyper selection, from hematopoietic cellular therapy to immunotherapy. Indications of these transplantations are now more clear for malignant diseases and are ongoing for auto-immune diseases. A better knowledge of the HSC allows the control of their proliferation and differentiation, opening the field of ex vivo expansion. Very recently, new stem cells have been identified, establishing that a differentiated cell retain its totipotency: a nervous system cell can differentiate into HSC, which will further give hematopoiesis, mesenchymental cells or hepatocytes. New tools are under development: human ES cells, biomaterials, functionalized materials, opening the field of cellular engineering in the year 2000. PMID- 10666795 TI - [Preoperative strategy for homologous blood salvage and peri-operative erythropoietin]. AB - The amount of transfused blood is related to blood loss calculated for the specific type of surgical procedure, transfusion hematocrit trigger and patient's red blood cell mass on the day before surgery. To optimise the benefit/cost and benefit/risk ratios of blood transfusion, a correct prescription must be done in accordance with the patient's red blood cell mass and surgical blood loss. Indeed, there is a clear need to define the appropriate uses of blood management methods and to seek new methods of improving perioperative blood management. The number of moderately anaemic patients undergoing surgery is currently thought to be 20%. Where transfusion requirements are estimated at two to three blood units, as for instance in the most common types of orthopaedic surgery, preoperative haemoglobin is the key factor governing transfusion needs. In this case, the simplest approach is to prescribe Epoetin Alfa subcutaneous at a dose of 600 IU/kg/week starting three weeks before the surgery. In addition, it is important in all cases to give concomitant iron supplements. Concomitant use of other methods to decrease allogeneic blood requirements is of no value. Obviously, the higher the haematocrit the day prior to surgery, the higher the patient's RBC mass and the greater the patient's permitted blood loss, decreasing the transfusion trigger. In this way, allogeneic blood loss is reduced, but without the need for the patient to attend the blood transfusion center and to undergo laboratory screening and testing of donated blood, and without the risk of inducing preoperative anaemia compared with sequential autologous blood donation. But, to optimise the benefit/cost ratio, we try to define precisely the patient populations likely to benefit from preoperative erythropoietin. Using different examples, management is proposed with algorithms. PMID- 10666796 TI - [Sensitivity of screening kits for anti-HIV antibodies. 1999 update. Retrovirus Working Group of the French Society for Blood Transfusion]. AB - A comparative evaluation of the sensitivity of anti-HIV screening assays has been recently performed with a selected panel of 65 samples which included HIV1 group M (per-seroconversions, seroconversions and seropositives infected with genotypes A, B, C, D, E), HIV1 group O and HIV2. The results obtained with the 21 ELISA HIV1 + HIV2 screening assays are presented. Among these 21 assays, four are combined anti-HIV and p24 Ag assays. All the assays except four fulfilled the criteria defined at the beginning of the study, i.e., positive results on all the seropositives including seroconversions and positive results on at least 50% of the per-seroconversions. The main differences were observed with the ten per seroconversion samples, the number of positive results on such samples varying from three to ten. The constant improvement of anti-HIV screening tests which leads one to shorten the 'window' period permits and earlier diagnosis of HIV infection and a progressive decrease of the transfusional risk. PMID- 10666797 TI - [Erythrocyte adhesion to the vascular endothelium]. AB - Blood cells are in continuous contact with the vascular endothelium. Endothelial cell culture, intravital videomicroscopy allowed the investigation of blood cell endothelium interactions in dynamic conditions. In the various diseases, diabetes mellitus, sickle cell anemia and malaria, erythrocytes have an increased adhesion to endothelial cells. The presence of advanced glycation end products (AGE) on erythrocytes of diabetics is responsible for their binding to the receptor RAGE present on the endothelium. The AGE-RAGE binding provokes an oxidant stress and induces the expression of the adhesion molecule. Furthermore, erythrocyte AGE induce an increase in vascular permeability. In sickle cell anemia, the increased adhesiveness and the sickling of red blood cells are responsible for thrombosis. Plasmodium falciparum infestation of erythrocytes induces knob formation at the cell surface and the P. falciparum protein binding to CD36, ICAM-1 and thrombospondin present on the endothelium, and facilitates the parasite dissemination. PMID- 10666798 TI - [Quality control of labile blood products. Why and howto properly take a specimen? Labile Blood Products Group of the French Blood Transfusion Society]. AB - This article reviews the various techniques of sampling used for the quality control of blood cell products. The importance of this stage for the validity of quality control results is emphasized. Three sampling methods, i.e., stripping, the sterile connection of sampling bag and the destructive method, are described in the form of operating modes and analyzed according to their advantages and drawbacks. The results of a comparative study carried out by the working group 'Blood Cell Products' of the French Society of Blood Transfusion are presented, showing that each method is valid and permits one to obtain a representative sample of the product to be controlled. Thus the diversity of the sampling methods allows us to select the one most adapted to the product to be controlled and to the analyses to be carried out afterward. PMID- 10666799 TI - [Preparation of cell therapy products: contribution of closed systems]. AB - A phase I clinical trial is being currently performed in our institution, aiming at evaluating the feasibility and toxicity related to the administration of Herpes Simplex-thymidine kinase gene-expressing human primary T lymphocytes following allogeneic hematopoietic stem cell transplantation. The need for safe and standardized preparation conditions for gene-modified cells is crucial. We describe the closed culture system used in the current trial for ex vivo retroviral-mediated gene transfer and transduced cell selection. Cell handling is performed in closed systems using sampling and transfer pack bags, culture bags and a sterile connection device which avoids opening the culture system. This closed system allows safe and reproducible ex vivo preparation of gene-modified primary T-lymphocytes for clinical use. PMID- 10666800 TI - Management of Lyme borreliosis and emerging tick-borne diseases in Europe. PMID- 10666801 TI - Principles of the diagnosis and antibiotic treatment of Lyme borreliosis. AB - Clinical signs and symptoms are an essential aspect of the diagnosis of Lyme borreliosis. Thus, a thorough knowledge of the clinical features of the disease is important. Established clinical definitions could be of help in everyday clinical practice and especially to compare the findings of different authors or groups. The characteristic sign that permits the diagnosis of Lyme borreliosis is a typical erythema migrans skin lesion. Highly suggestive manifestations are ear lobe lymphocytoma, acrodermatitis chronica atrophicans and Bannwarth's syndrome. The majority of other signs and symptoms are only suggestive and, when expressed individually, may have a very limited or even symbolic value for the purpose of diagnosis. Laboratory confirmation of borrelial infection is needed, as a rule, for all manifestations of Lyme borreliosis with the exception of typical erythema migrans. In clinical practice indirect laboratory methods are usually employed. Determination of borrelial IgM and IgG antibodies by immunofluorescence assays or enzyme-linked immunosorbent assays has not been standardised, and correlation of the results from different laboratories and/or different commercial tests may be poor. Immunoblotting may solve some of the many dilemmas but could (especially in Europe) raise additional questions in a field in which numerous uncertainties already exist. The reliability of methods for direct detection of borrelial infection other than culture to ascertain spirochetes in tissue specimens is open to question. Treatment with antibiotics is reasonable in all stages of Lyme borreliosis and for all clinical manifestations; however, it has been most effective early in the course of the illness. The choice of antibiotic depends upon many factors including the efficacy, pharmacokinetic profile, side effects, expected compliance and price. For the majority of manifestations the most effective antibiotic, the optimal dosage, and the most appropriate duration of treatment have not been exactly determined. Recommendations for the treatment of Lyme borreliosis in Slovenia are presented. PMID- 10666802 TI - Comparison of cefuroxime axetil and phenoxymethyl penicillin for the treatment of children with solitary erythema migrans. AB - OBJECTIVE: To compare the clinical efficacy and drug-related adverse effects of 14 days of treatment with cefuroxime axetil 30 mg/kg/day or phenoxymethyl penicillin 100,000 IU/kg/day in the treatment of children with erythema migrans. METHODS: Consecutive patients younger than 15 years, referred to our institution in 1996 with solitary erythema migrans and without prior antibiotic therapy, were included in this prospective study. Basic demographic features and clinical data were collected by questionnaire. The efficacy of the treatment of acute disease, development of major and/or minor manifestations of Lyme borreliosis and drug related adverse effects were surveyed at follow-up visits during the first year after the initiation of antibiotic treatment. RESULTS: Forty-six patients received cefuroxime axetil (group C) and 44, phenoxymethyl penicillin (group P). The two groups differed in terms of age (patients in group C were younger), but no other differences in demographic and clinical pre-treatment characteristics were present. The clinical course during the post-treatment period revealed no significant differences between the two groups: the duration of erythema migrans (7.1 +/- 7.5 days in group C, 10.6 +/- 19.3 days in group P) and the appearance of minor manifestations of Lyme borreliosis (8.8% in group C, 9.1% in group P) were comparable; no major manifestations were recorded. Twelve months after antibiotic treatment all patients were free of symptoms. The patients treated with cefuroxime axetil had more drug-related adverse effects than did those treated with phenoxymethyl penicillin (26.1% versus 6.8%, p = 0.0301). "Herxheimer's reaction" at the beginning of treatment was identified more often in group C than in group P, but the difference was not statistically significant. CONCLUSIONS: Cefuroxime axetil and phenoxymethyl penicillin are equally effective in the treatment of children with solitary erythema migrans; however drug-related adverse effects were more frequently observed with cefuroxime axetil. PMID- 10666803 TI - Erythema migrans in the immunocompromised host. AB - From 1990 to 1996 a total of 67 adult patients with typical erythema migrans (EM) and a previously identified immunocompromised condition were investigated at the University Medical Centre, Department of Infectious Diseases, Ljubljana, Slovenia. The course and outcome of borrelial infection were compared with 67 previously healthy age and sex-matched individuals with EM who were examined at our institution in the same year. Clinical characteristics of Lyme borreliosis before treatment and the duration of EM after the institution of therapy with antibiotics including amoxicillin, azithromycin, cefuroxime-axetil, doxycycline, and ceftriaxone were comparable in both groups. The occurrence of early disseminated borrelial infection before treatment and the frequency of treatment failure (defined as the onset of severe minor or major manifestations of Lyme borreliosis, persistence of B. burgdorferi sensu lato in the skin and/or persistence of EM after treatment) were found significantly more often in immunocompromised patients than in the control group (16/67 versus 6/67, respectively; p = 0.0358). Re-treatment was required in 13 (19.4%) patients of the immunocompromised group and only in five (7.5%) patients of the control group (p = 0.0762). However, in spite of the more severe course and the more frequent need for re-treatment among patients whose immune system was impaired, the outcome of borrelial infection after one year was favourable in both groups. PMID- 10666804 TI - Erythema migrans in pregnancy. AB - From 1990 through to 1997, 105 pregnant women with typical EM were investigated at the Lyme Borreliosis Outpatients' Clinic of the Department of Infectious Diseases at the University Medical Centre in Ljubljana, Slovenia. Twenty-five (23.8%) patients acquired borrelial infection during the first trimester of pregnancy; eight (7.6%) of them had noticed the skin lesion before they became pregnant. In 43 (40.6%) patients the EM appeared in the second trimester, and in 37 (35%) patients in the third trimester of pregnancy. Two (1.9%) patients received phenoxymethyl penicillin (1 million IU t.i.d.), three (2.9%) benzyl penicillin (10 million IU b.i.d.), and 100 (95.2%) ceftriaxone (2 g daily). All patients were treated for 14 days except three (2.9%) in whom the treatment with ceftriaxone was discontinued because of mild side effects. The outcome of disease was good in all 105 patients. Ninety-three (88.6%) out of 105 patients had normal pregnancies; the infants were delivered at term, were clinically healthy, and subsequently had a normal psychomotor development. In the remaining 12 (11.4%) patients an adverse outcome was observed. Two (1.9%) pregnancies ended with an abortion (one missed abortion at 9 weeks, one spontaneous abortion at 10 weeks), and six (5.7%) with preterm birth. One of the preterm babies had cardiac abnormalities and two died shortly after birth. Four (3.8%) babies born at term were found to have congenital anomalies; one had syndactyly at birth and three had urologic abnormalities which were registered at the age of 5, 7, and 10 months, respectively. A causal association with borrelial infection was not proven in any infant. For at least some unfavourable outcomes a plausible explanation not associated with Lyme borreliosis was found. PMID- 10666805 TI - The outcome of Lyme borreliosis in children. AB - Austria is an endemic area for Lyme borreliosis. The IgG seroprevalence of healthy blood donors as investigated by a DAKO flagellum-ELISA in Graz/Styria is 13%. In order to determine whether this high seroprevalence is caused by infection in childhood, 36 children aged 3 to 18 years (mean, 10.1 years) were followed up over 2 to 20 months (mean, 11.1 months) and reinvestigated for clinical symptoms and antibodies against B. burgdorferi by a commercial flagellum ELISA and a commercial B. garinii Western blot (WB). Twenty-seven children had erythema migrans (EM), one of them with reinfection, 5 had borrelia lymphocytoma (BL), 2 EM and BL, 1 acrodermatitis chronica atrophicans and 1 ACA/circumscribed scleroderma. Before treatment with either phenoxymethylpenicillin, amoxicillin, or minocyclin for 3-4 weeks, 64% of the patients were IgM and 44%, IgG seropositive. Clinically, all but 5 patients with EM recovered from Lyme borreliosis. Among these 5 patients--one of them with reinfection of EM--3 had mild arthralgia, 1 recurrent headache and 1 concentration disturbance. Only 2 children with arthralgia were IgM positive by ELISA and WB. One of 5 BL patients had a persistent swelling of the ear lobe although with a negative serology before and after several antibiotic treatments and at follow up. In 16 children serological investigations were performed after more than 12 months (range, 13-20 months). Eighteen percent of them had IgM antibodies by ELISA, 25% by WB, and 6% IgG antibodies by ELISA and 6% by WB. Although there was a decline of antibody response from 64% to 18% for IgM and from 44% to 6% for IgG as measured by ELISA, children remain seropositive for more than 1 year with or without clinical symptoms. The relevance of the association with clinical symptoms can be raised by combining several diagnostic methods. It is assumed that recurrent, often silent, infections might increase antibody titres. It should be noted that antibody titres also generally increase with the age of individuals. PMID- 10666806 TI - Has the presence or absence of Borrelia burgdorferi sensu lato as detected by skin culture any influence on the course of erythema migrans? AB - The aim of this prospective study was to compare epidemiological and clinical data in patients with a positive Borrelia burgdorferi sensu lato culture and culture-negative erythema migrans skin lesions. Of the 546 adult patients with erythema migrans seen at our institution in 1997 in whom a skin biopsy was performed and the specimen cultured for the presence of B. burgdorferi sensu lato, 235 (43%) had a positive and 311 (57%) a negative skin culture. More women than men were present in both groups and women were also significantly older than men. Tick bites resulting in culture-positive erythema migrans predominated in May (p = 0.012), while in August and September tick bites with subsequent culture negative skin lesions were more common (p = 0.018 and 0.011, respectively). Similarly, erythema migrans lesions noticed by our patients in May were significantly more often Borrelia culture positive than negative (p = 0.004), while lesions appearing in October were significantly more often culture negative (p = 0.004). In addition to these seasonal differences, the comparison of the large number of Borrelia skin culture-positive and -negative patients with erythema migrans also revealed differences in several clinical parameters including a larger diameter of skin lesions in the culture-positive group (p = 0.007 at presentation, and p = 0.039 at registration, respectively), a lesser number of multiple skin lesions (7/235 versus 27/311, p = 0.006), and a lower frequency of signs/symptoms (p = 0.039) associated with erythema migrans lesions in culture-positive than in culture-negative patients. We have no plausible explanation for the majority of these rather unexpected findings. Of the 59 patients who, prior to biopsy, had received brief courses of antibiotics known to be effective in the treatment of erythema migrans, 12 (20.3%) were culture positive. As anticipated, the ratio of culture positivity in pretreated patients was significantly lower (p < 0.001) than in those without antecedent antibiotic therapy. PMID- 10666807 TI - Erythema migrans and serodiagnosis by enzyme immunoassay and immunoblot with three borrelia species. AB - There is wide divergence of opinion between physicians regarding the use of serological measures for the diagnosis and treatment of erythema migrans, the hallmark of Lyme borreliosis. We studied the outcome of an enzyme immunoassay and immunoblot (Western blot) used on the sera of patients who had suffered tick bite and erythema migrans, and had been subsequently treated with various antibiotics. Ninety-nine consecutive patients presenting with erythema migrans after tick bite were prospectively recruited at the outpatient department of two Vienna City hospitals and at the consultation office for Lyme borreliosis of the Institute of Hygiene. University Vienna. Blood samples were taken before antibiotic treatment and 3 and 6 months thereafter. Blood samples from 100 blood donors served as controls. Antibodies against Borrelia burgdorferi sensu lato were determined by enzyme immunoassay (IgG and IgM EIA) and by IgG immunoblot. The latter was performed with isolates of B. alzelii (H2) B. burgdorferi sensu stricto (Le) and B. garinii (W) from Austrian patients. The 4 interpretation criteria for immunoblot results were: A (3 bands out of 8), B (2 bands out of 9), C and D (1 band out of 6). In all patients, the erythema resolved within the treatment period. No complications secondary to the borrelia infection were registered. After treatment there was no significant change in titre, nor was there a difference in the immunoblot pattern between the first, second and third serum samples. Serum antibodies to B. burgdorferi were positive by EIA in 22.9% (IgG) and 2.5% (IgM). Immunoblot results offered by borrelia species and by the interpretation criteria, ranging between 8.3% (criterion A, strain Le) and 44.2% (criterion D, strain H2). By EIA, control samples were IgG and IgM positive in 5% and 1%, respectively. Positive immunoblot results with strain H2 were found in 9%, 13%, 18%, and 20% by the criteria A through D respectively. After antibiotic treatment of erythema migrans the immunological response appears to be abrogated. Thus, serological results are not supportive for the diagnosis of erythema migrans, not will they retrospectively prove successful antibiotic treatment of borrelia infection. PMID- 10666808 TI - Is IgM of diagnostic value in case of delayed intrathecal production of IgG antibodies? AB - The neurological manifestations of Lyme borreliosis comprise a wide range of clinical signs. However, these symptoms might have other aetiologies. Therefore detection of intrathecal production of specific antibodies is necessary to confirm the clinical assumption of neuroborreliosis (NB). In case of delayed intrathecal production of specific IgG antibodies, detection of IgM could play a role in the early diagnosis of NB. To clarify whether IgM is of diagnostic value in such cases, paired CSF serum samples from 176 patients with suspected NB admitted to the department of Neurology, Karl Franzens University, Graz, Austria, were tested. Testing was performed with the IDEA Neuroborreliosis Kit (Dako, Denmark) and Enzygnost Borreliosis (Behring, Germany) and results of both methods were compared. According to well defined criteria 63 of the 176 patients had defined NB and 113 were regarded as possible NB. Twelve out of 63 patients with defined NB had delayed intrathecal IgG production. Only one patient with delayed IgG production had an intrathecal IgM production prior to IgG. In all patients with possible NB no intrathecal production of IgM was detected. At the time of the first lumbar puncture IgG intrathecal production could be detected with the IDEA seven times more often than with the Enzygnost Borreliosis. The determination of intrathecal production of IgM does not appear to be of diagnostic value in patients with delayed IgG antibody production. Therefore a consecutive lumbar puncture is more likely to confirm clinical assumption if there is strong clinical evidence of NB. PMID- 10666809 TI - Lyme meningitis: a one-year follow up controlled study. AB - Thirty-six patients with Lyme meningitis diagnosed at the Department of Infectious Diseases, University Medical Centre, Ljubljana in 1993 and 1994 were enrolled in a prospective study. All patients had lymphocytic meningitis, negative serum IgM antibody titres to tick-borne encephalitis virus and met at least one of the following four criteria: i) isolation of Borrelia burgdorferi sensu lato from cerebrospinal fluid (2 patients), ii) intrathecal borrelial antibody production (22 patients) iii) seroconversion to borrelial antigens (3 patients) and/or iv) erythema migrans in the period of four months prior to the onset of neurological involvement (21 patients). All patients underwent antibiotic treatment and were followed up for one year. The results of our study revealed that Lyme meningitis frequently occurs without meningeal signs and is often accompanied by additional neurological and/or other manifestations of Lyme borreliosis. During the first year after antibiotic treatment, minor and major manifestations of Lyme borreliosis persisted or occurred for the first time in several patients. They were not infrequent even at the examination performed one year after therapy. PMID- 10666810 TI - Borrelia burgdorferi as a cause of Morgagni-Adams-Stokes syndrome. Long time follow-up study. AB - According the literature atrio-ventricular blockade (AVB) is the most frequent and well-known symptom of Lyme carditis. Typical signs of complete AVB include fatigue, lethargy and syncope- Morgagni-Adams-Stokes syndrome (MAS). The authors present their results and experience with 5 patients selected from a long-term study (conducted between 1987 and 1998) comprising 58 patients who developed MAS. The authors tried to evaluate the changes especially in the cardiovascular system. They correlated the clinical state with ECG findings, as well as with the levels of the Borrelia burgdorferi antibodies. The following results were obtained: 1) all patients had typical syncope, 2) the clinical course was not complicated (except one patient who developed ventricular fibrillation), 3) two patients had frequent symptomatic and asymptomatic arrhythmia including chest pain and episodic rest dyspnea, 4) subjective difficulties (usually palpitations) correlated with ECG findings (Lown 3a, 3b). The authors also looked for any relationship between clinical difficulties and levels of antibodies. The results obtained with an early permanent pacemaker were less favourable than those reported in the literature. Despite early treatment 2 patients had repeated palpitations and ECG correlates during the next years. PMID- 10666811 TI - Lyme borreliosis and peripheral facial palsy. AB - From 1994 to 1996, 114 consecutive patients older than 15 years who presented at the Department of Infectious Diseases, University Medical Centre, Ljubljana, fulfilled the criteria for inclusion into this study on the borrelial aetiology of peripheral facial palsy (PFP). The study was restricted to patients without a conceivable explanation for their PFP, erythema migrans or history of erythema migrans, clinical signs/symptoms of frank meningitis or any other neurological manifestation in addition to PFP. In 22 (19.3%) of these 114 patients borrelial infection was confirmed by one of the following: in 3 (13.6%) by the isolation of Borrelia burgdorferi sensu lato from cerebrospinal fluid (CSF), in 11 (50%) by the presence of intrathecal antibody production, and in 8 (36.4%) by seroconversion to borrelial antigens. Additional 20 (17.5%) patients interpreted as having had a probable borrelial infection, had positive (> or = 1:256) IFA IgM and/or IgG borrelial serum antibody titres, and in 9 (7.9%) patients borderline borrelial antibody titres (1:128) were found (interpreted as a possible infection). In 63 (55.3%) patients the serological tests remained negative. Lymphocytic pleocytosis was found at the first visit in 12/22 (54.5%) patients with confirmed borrelial infection, in 3/20 (15%) with probable infection, in 1/9 (11.1%) with possible infection, and in 10/63 (15.9%) patients with symptoms of unknown aetiology. Patients with confirmed borrelial infection had abnormal CSF findings significantly more often than did patients with symptoms of unknown aetiology (p = 0.0139 for lymphocytic pleocytosis and/or elevated CSF protein levels, and p = 0.0010 for lymphocytic pleocytosis). Local and systemic signs/ symptoms were also more common in patients with confirmed borrelial infection than in those with an symptoms of unknown aetiology (p = 0.0258). In Slovenia which is a highly endemic region for Lyme borreliosis, borrelial infection is a frequent cause of PFP in adult patients. PFP may occur early in the course of LB, prior to measurable antibody response, indicating the need for serologic follow up. Abnormal CSF results and the presence of additional local and/or systemic symptoms are factors indicating a higher possibility of borrelial aetiology of PFP and should alert physicians to suspect LB. PMID- 10666812 TI - Diffuse reversible alopecia in patients with Lyme meningitis and tick-borne encephalitis. AB - Alopecia occurring after febrile bacterial and viral infection is a phenomenon well known since the beginning of the century. To evaluate the occurrence of alopecia in tick transmitted disease, 23 adult patients with Lyme meningitis and 71 patients with tick-borne encephalitis were included in a prospective study and were followed up for one year. Diffuse alopecia occurred within three months after the outbreak of disease in 3 out of 23 (13%) patients with Lyme meningitis and in 40 out of 71 (56.3%) patients with tick-borne encephalitis. The mean duration of alopecia was 2 to 3 months and alopecia was reversible in all patients. PMID- 10666813 TI - The course of Lyme disease in different age groups. AB - Lyme disease is a multisystem infection affecting all age groups. In this study an attempt was made to determine whether the patient's age influences the course of the disease. One hundred and fifty patients with diagnosed Lyme disease were included in the study. Two serological methods were used to detect antibodies to Borrelia burgdorferi and to confirm the diagnosis: an indirect immunofluorescence assay (the Russian strain Ip-21) and Western blot. The course of Lyme disease did not differ from that seen in Europe and North America. However, a few clinical differences between groups were observed. In the first age group (0-15 years) the most common manifestation was flu-like symptoms with fever. Neither radiculoneuritis nor polyneuropathy was observed in this age group. Late manifestations were rare and the outcome of the disease was benign. The course of the disease in the second group (16-40 years) was most similar to that in childhood and the also outcome was similar. Carditis and erythema multiple were significantly more common in the second group (16-40 years) than in the other age groups. No differences were found between the third (41-60 years) and fourth (> than 60 years) group in the frequency of flu-like symptoms, erythema multiple and aseptic meningitis. However, the most important clinical sign in this group was involvement of the nervous system whereas in the third group this was joint damage. This feature deserves attention because, as a rule, the presence of an articular syndrome determines the prognosis of LD. PMID- 10666814 TI - Studies on the pathogenesis and treatment of Lyme arthritis. AB - Lyme arthritis is one of the most common clinical manifestations of Lyme borreliosis. It is caused by an intraarticular infection with Borrelia (B.) burgdorferi. A small number of bacteria are liable to provoke severe arthritis by inducing mechanisms (including the induction of cytokines and chemokines) that amplify the inflammatory response. The cellular immune response against B. burgdorferi is characterised by a predominant T helper cell type 1 (Th1) pattern that appears to be inadequate to overcome the infection. In most cases, Lyme arthritis may be cured by antibiotic therapy. A brief summary of current recommendations for the treatment of Lyme arthritis in adults and children is given in this article. However, about 10% of Lyme arthritis patients do not respond sufficiently to antibiotic treatment. Two not mutually exclusive pathogenetic concepts of these treatment-resistant cases will be discussed in the present study: persistent infection and infection-induced immunopathology. PMID- 10666815 TI - Functional heterogeneity in the antibodies produced to Borrelia burgdorferi. AB - Antibodies to outer surface molecules of Borrelia burgdorferi (Osp) that have a bactericidal action in the absence of complement have been described. These antibodies are primarily monoclonal to antigenic determinants in OspA and OspB. One of these, CB2, is an IgG1 monoclonal antibody that recognizes an epitope in the carboxy terminus of OspB. The specificity of CB2 is critically dependent on the presence of a lysine (Lys) residue in position 253, not only for binding but also for killing the spirochete. This antibody has been used successfully to select escape variants or mutants that are missing the Lys residue either by a mutation or by deletion as a result of premature stop codons. Other antibodies to OspA, OspB, and p39 have also been characterized with similar properties. Another important feature of CB2 is that its bactericidal action is not dependent on agglutination, since Fab fragments of the whole immunoglobulin molecule can also kill in the absence of complement synergy. The killing action of CB2 is not inhibited by protease inhibitors, and is dependent on the presence of calcium. Upon contact with Borrelia burgdorferi, CB2 causes lysis of the outer membrane and the formation of a spheroplast. The bactericidal mechanism of this antibody is not known. The sequence of the heavy and light chain variable regions of CB2 have striking homology to murine antibodies of the autoimmune repertoire, and some of these antibodies have catalytic properties. In general, catalytic antibodies have enzymatic rates of acceleration that are significantly less than those of proteolytic enzymes. If CB2 were a catalytic antibody, its substrate specificity may be expected to be broader. CB2 does not cleave recombinant OspB, nor does it cleave other protein substrates. Its killing activity is not dependent on proteolysis. Because the bactericidal action of CB2 involves the destruction of the outer membrane, it is possible that a phospholipase could be associated with the mechanism. The mobility of spirochetal lipids is altered after incubation with CB2, and the bactericidal activity is reduced in the presence of phospholipase inhibitors. These studies suggest that the bactericidal mechanism of CB2, and other similar antibodies, is novel. PMID- 10666816 TI - Risk assessment in Lyme borreliosis. AB - A 3-year EU-funded project (EUCALB), initially involving 14 countries and more than 30 scientists and physicians, was undertaken with the main objective of identifying practical risk assessment criteria for Lyme borreliosis. A major part of the project was dedicated to the improvement of data quality. European case definitions were formulated and quality assurance schemes were developed for serological diagnosis and the detection of infection in ticks. Studies on the standardisation of immunoblot interpretation criteria are still in progress. Data on the clinical risk from tick bites were obtained and considerable progress was made in elucidating the complex ecology of the disease. A study on habitat assessment throughout Europe concluded that high risk was associated with highly heterogeneous recreational woodland and case data from both high and low incidence countries suggested that most infections were acquired in recreational areas. Considerable work is still required to relate clinical data to the epidemiology and ecology of the disease in order to assess risk in Lyme borreliosis. PMID- 10666817 TI - Species identification of Borrelia burgdorferi sensu lato from tick and human isolates in Styria (Austria) by PCR-RFLP analysis. AB - Seventy-one isolates of Borrelia burgdorferi sensu lato (B.b.s.l.) derived from Ixodes ricinus ticks (50 strains) and patients (21 strains) were characterised by PCR-RLFP analysis. In four cases the human isolates were obtained from the cerebrospinal fluid (CSF) of patients with clinical symptoms of neuroborreliosis and in 17 cases from skin biopsies of patients with dermatological manifestation of Lyme borreliosis. Ixodes ricinus isolates originated from 14 localities in three regions (Mur valley, eastern and western Styria) in Styria. Thirty six strains of B.b.s.l. were isolated from nymphal ticks, nine strains from female and five strains from male ticks. Species identification of human isolates revealed three B. garinii and one B. afzelii isolates in CSF. In the PCR-RFLP analysis of 17 skin specimens a pattern for B. afzelii was found in ten cases, while six could be identified as B. garinii and one as a mixed infection of B. afzelii and B. garinii. Genetic characterisation of tick isolates resulted in 24 strains of B. afzelii (48%), 11 strains of B. garinii (40%) and 5 strains of B. burgdorferi s.st. (10%); one isolate showed a mixed infection of B. afzelii and B. garinii. Our findings indicate that B. afzelii and B. garinii predominate over B. burgdorferi s.str. in Ixodes ricinus ticks from Styria, which is similar to findings in neighbouring countries. This also reflects the occurrence of different pathogenic Borrelia strains in human samples. PMID- 10666818 TI - Infection of small mammals with Borrelia burgdorferi sensu lato in Slovenia as determined by polymerase chain reaction (PCR). AB - Thirty-four small mammals collected in the vicinity of Ljubljana were tested for the presence of Borrelia burgdorferi sensu lato by polymerase chain reaction (PCR) of urinary bladder tissues, using universal flagellin primers and species specific rRNA primers. Seventeen small mammals (50%) were found to be positive, and 7 small mammals were infected with two species of B. burgdorferi sensu lato simultaneously. The most commonly found species was B. afzelii (n = 14), followed by B. burgdorferi sensu stricto (n = 7) and B. garinii (n = 3), as determined by species-specific primers. We conclude that PCR is a rapid and reliable method to detect infection with B. burgdorferi sensu lato in small mammals. PMID- 10666819 TI - Epidemiological aspects of human granulocytic Ehrlichiosis in southern Germany. AB - Human granulocytic Ehrlichiosis (HGE) is a newly emerging acute febrile illness which is likely transmitted by ticks of the Ixodes ricinus/I. persulcatus complex. First seroepidemiological surveys on the prevalence of HGE antibodies, detection of DNA of granulocytotropic Ehrlichiae in I. ricinus and one case of HGE from Slovenia confirmed by serology and PCR (polymerase chain reaction) suggest that HGE might exist all over Europe. The purpose of the present study was a) to determine the prevalence of antibodies against the HGE agent in sera collected from persons at high risk for exposure to I. ricinus with that of a control population and b) to determine the prevalence of granulocytic Ehrlichiae in I. ricinus ticks from Southern Germany. We studied sera from 150 forestry workers and 105 patients with an established diagnosis of Lyme disease as tick exposed populations. Sera from 103 healthy blood donors without a history of known tick bites served as controls. A significantly higher prevalence of HGE antibodies (P < or = 0.01) was present among patients with Lyme borreliosis (12 of 105 were positive; 11.4%) and forestry workers (21 of 150 were positive; 14%) compared to blood donors (2 of 103 were positive; 1.9%). Furthermore, 510 adult and nymphal I. ricinus were investigated by PCR for the presence of granulocytic Ehrlichiae with primers specific for the E. phagocytophila group. In eight (1.6%) of the investigated ticks the expected amplification product was detectable, indicating a low prevalence of infected ticks especially when compared with B. burgdorferi. The presented data strongly suggests that the HGE agent or a closely related organism exists in Southern Germany and therefore HGE should be considered in the differential diagnosis of febrile illnesses. However, final evidence can be provided only after isolation of the organism from patients. PMID- 10666820 TI - [Reducing pain by oral enzyme therapy in rheumatic diseases]. AB - Proteolytic enzymes have analgesic, effects, besides the wellknown antiinflammatory and edema-reducing properties. These analgesic effects are based on the inhibition of inflammation and in addition to that on direct influences on the nociceptors. All that explains the therapeutical effects of such enzymes in degenerative-rheumatic and soft tissue rheumatic diseases in which inflammatory or immunologic processes are not in the forefront. In recent years a significant reduction of pain in various rheumatic diseases, concerning these aspects, was shown in several clinical studies. The clinical trial in patients with periarthritis of shoulder showed statistical equivalence of pain reduction, whether they were treated with phlogenzym or diclofenac. Likewise in the trial of patients suffering from painful osteoarthritis of the knee, there was a statistical equivalence of the pain-scores, comparing diclofenac and enzymes. The study of painful vertebral syndromes again resulted in equivalence of the treatment with NSAIDs compared to therapy with enzymes. PMID- 10666821 TI - [Compression treatment after burns]. AB - After healing up of the injury wounds, hypertrophic scars and keloids often develop, which are histologically characterised by irregulary arranged collagen fibre bundles and a strong vascularisation. Approximately 20 years ago, the so called compression clothing, as for example suits, masks, gloves, stockings, were first employed for the prevention and therapy of these complications. These means of compressions are crosswise and lengthwise elastical and consist predominantly of elasthan and viscose. The pressure acting on the skin lies between 25 and 32 mmHg: thereby the values are above the average capillary pressure of 20 mmHg. The efficiency of the compression clothing after a burn injury is well proved by several studies, and one knows today that, for example in the case of children as from the 5th year of life, the results are better than in the case of adults from the 35th year of life. The compression effected at least during a period of 15 months slows down the blood circulation, reduces the number of capillaries and makes the scar become more pale. Furthermore, the orthologically parallel arranged collagen fibres maintain their arrangement due to the compression pressure and do not get irregularily arranged. PMID- 10666822 TI - [Alzheimer disease--differential diagnosis and modern therapy]. AB - Alzheimer's disease and other dementias are a multidisciplinary challenge and become more and more important because of the growing life expectancy, and also due to the presence in the media. A. Alzheimer published first 100 years ago the disease named after him as an entity marked by the clinical course and by typical histological findings. The last decades brought remarkable progress in demarcation to other forms of dementias for example by CT or MRI. The cause of Alzheimer's disease has not yet been cleared up. The finding of secondary dementias has great importance since causal therapeutic possibilities exist. In the last years new ideas to etiology in respect to the hypothesis of acetylcholine resulted in new therapeutic promises. A rational treatment of this disease seems now possible. Besides psychological and behavioral signs and symptoms of dementia are of great importance in the care of these patients. PMID- 10666823 TI - [Therapy of heart failure with ACE inhibitors--"evidence-based medicine" and clinical reality]. AB - Treatment of patients with chronic heart failure improves symptoms and NYHA functional class in about 50-80% of all patients treated. A 15% reduction can be observed in the need for hospitalisation and a 16-31% reduction in 1-year mortality. 37% risk reduction for progression to symptomatic heart failure can be achieved with ACE-inhibition in asymptomatic patients with systolic left ventricular dysfunction (55). Thus, ACE-inhibition should be part of standard treatment in symptomatic and asymptomatic patients with left ventricular dysfunction. In symptomatic patients with chronic heart failure, combination therapy of an ACE-inhibitor with digitalis and diuretics is state of the art and improves symptoms significantly. The addition of a vasodilatator can be considered in selected cases. Based on recently published data on beta adrenoreceptor-blockade in the treatment of chronic heart failure, beta-blockers seem to get part of standard therapy of heart failure in the present and near future (2, 44, 50, 60). In spite of innovations of modern heart failure therapy prognosis is still bad. Survival after diagnosis of severe heart failure (functional class NYHA III and IV) is limited to a mean of 14 month. PMID- 10666824 TI - [Peripheral arterial occlusive disease]. PMID- 10666825 TI - [Methods and results of controlled walking training in patients with peripheral arterial occlusive disease]. AB - To establish the effect of exercise training in patients with PAOD Stage II according to Fontaine, 125 publications between 1966 and 1998 were systematically reviewed. The best results occurred with the following method: duration greater than 30 minutes per session, frequency of at least three sessions per week, intermittent walking to near-maximal pain and program length of greater than six months. A methodological study of six randomized clinical trials was performed. The claudication distance was assessed with a treadmill test at a constant-load of 0.82-1.06 Watt/kg. Improvement in initial/absolute claudication distance (ICD/ACD) ranged from +28% to +213%. The average increase was +150% in ICD, and +200% in ACD. Exercise rehabilitation significantly improved patient's quality-of life as well. However, the benefit of exercise training however was only moderate (6-32%). Exercise training is a very effective therapeutic measure. However, for further use better guidelines and prospective cost-effectiveness studies including evaluation of rehabilitation programs with regard to improvements in functional capacity, modification of risk factors, long-term compliance, quality of-life and medical costs incurred are needed. PMID- 10666826 TI - [Antithrombotic therapy and follow-up of peripheral arterial occlusive disease]. AB - In patients with arterial occlusive disease the following antithrombotic drugs are used to prevent new arterial thromboses: Unfractionated heparin (UFH) and low molecular weight heparins (LMWH): UFH is frequently used perioperatively in patients with vascular reconstructions and after peripheral percutaneous transluminal angioplasty (PTA). Only one study on the longterm use of a LMWH after operative revascularization has been published, where LMWH led to an increased patency rate in comparison to acetylsalicylic acid + dipyridamol. Vitamin-K-antagonists were effective in the prevention of new vascular occlusions in studies published in the 1970ies. Platelet function inhibitors (aspirine, ticlopidine and clopidogrel): Aspirine reduced reocclusions after vascular surgery and PTA. Ticlopidine was effective in the Swedish Ticlopidine-Multicenter Trial in the prevention of ischemic heart disease and also reduced the need for vascular surgery. In the CAPRIE-study clopidogrel reduced myocardial infarctions in patients with peripheral arterial disease in comparison to aspirine. PMID- 10666827 TI - [Value of vasoactive drugs in conservative therapy of peripheral arterial occlusive disease]. AB - Based on controlled clinical trials, vasoactive drugs (pentoxyfylline, naftidrofuryl, buflomedil) and prostanoids (alprostadil, iloprost) are used in symptomatic peripheral arterial occlusive disease: In stage II, vasoactive drugs are prescribed in order to improve leg hyperemia in response to muscular work; in complicated stage II (e.g., non-healing traumatic skin lesions), and in stages III/IV, prostanoids are given in order to increase forefoot skin blood flow. These drugs should only be applied if neither angioplasty nor vascular surgery are recommendable, possible, or successful after complete angiological diagnostic. It should be considered that these substances have a rather limited clinical efficacy in both stage II and stages III/IV and therefore attempts should be made to select patients with a reasonable chance to benefit. These drugs are also not suitable for primary or secondary prophylaxis against the progression of vascular lesions. PMID- 10666828 TI - [Special aspects of therapy of non-atherosclerotic vascular diseases]. AB - The long term course of thromboangiitis obliterans as well as frequency and extent of major or minor limb amputations depend almost exclusively on the smoking behaviour of the patients. Superficial phlebitis accompanying an acute relapse responds well to high-dose aspirin or NSAIDs. Critical limb ischemia is treated by intra-arterial or intravenous prostaglandins (Alprostadil, Iloprost). Lokal measures for finger, toe, or foot gangrene do not differ from comparable sequelae of atherosclerotic vascular disease. Revascularisation procedures (angioplasty, surgery) have a high rate of technical failure and are indicated only in rare atypical situations. Corticosteroids are the therapy of choice for both vasculitides of large muscular arteries, i.e. temporal arteritis (M. Horton) and Takayasu arteritis. Combination therapy is restricted to steroid refractory disease; while this is the exception in temporal arteritis, it occurs in up to 50% of patients with Takayasu arteritis. Critical limb ischemia due to giant cell arteritis may persist even if the inflammatory activity of the disease is well controlled. Revascularisation procedures in Takayasu arteritis may have good results; as with all other therapeutic measures in this disease, they should be provided by specialized centers. Treatment of Raynaud's phenomenon requires patient evaluation for signs or symptoms of an underlying disease, i.e. some kind of connective tissue disease. Strength and frequency of attacks depend on a number of different factors (triggers) which in a given patient may not be completely understood. Exposition prophylaxis for known triggers and vasodilator drugs are the main therapeutic measures in Raynaud's phenomenon. Careful documentation of disease activity provided, non-classical remedies (behavioural psychotherapy, acupuncture) may be attempted. PMID- 10666829 TI - [Fibrinolytic, revascularizing therapy of peripheral arterial occlusive disease]. AB - Thrombo-embolic events are the primary or secondary cause for occlusions of peripheral arteries. The systemically applied fibrinolysis rarely resulted in revascularization. The local application of plasminogen activators leads to better results. Combined with interventionally techniques 70-80% of the acute und subacute occlusions can be opened an the necessity of angio-surgical interventions can be reduced by about 50%. The technical procedure of the local fibrinolysis and the dosages of the applied activators are presently not standardized. Therefore the results of 13 reports between the years 1995-1998 including 4 prospective studies are not comparable. According to the present experiences, acute und subacute incomplete ischemia syndromes are to be seen as indication for local lysis. Especially for patients whose constitution is not optimal for surgery, local fibrinolysis is an alternative to surgical revascularization. Further studies are necessary to optimize the therapy. PMID- 10666830 TI - [Possibilities and limits of interventional therapy in chronic peripheral arterial occlusive disease]. AB - In the therapy of the chronic peripheral vascular occlusion, angioplasty is rarely used to treat the infra-renal aortic stenosis, whereas the stenosis and the short occlusion of the iliac artery is a classical indication. Primarily, stenoses and occlusions of the iliac artery should be treated with balloon angioplasty exclusively. Only secondarily, when the result of angioplasty was insufficient, e.g. remaining stenosis or dissection, stent implantation is appropriate. Angioplasty is most frequently applied in the obliteration of the femoro-popliteal artery. It can be stated that early- and long-term results are the better, the shorter the occlusion is. Stent implantation in the femoro popliteal artery should be avoided because of poor results. Only with the intention of limb salvage, when there is no opportunity for surgical treatment, a stent implantation should be considered. In case of recurrent stenoses after stent implantation, angioplasty can be reapplied with great success. The indication for any vascular intervention should be a decision of both, the interventional radiologist and the vascular surgeon, because both kinds of treatment are palliative and not causal. It is the task of the angiologist to do clinical diagnostics and the after-treatment. Quality monitoring is indispensable. It consists of documentation of pre-angioplasty diagnostics and should be able to prove the correct indication for the intervention. The result of the intervention should also be documented by angiography and functional tests. Regular control of the patient after the intervention is necessary for the early recognition of recurrent stenoses. PMID- 10666831 TI - [Current status of vascular surgery]. AB - German vascular surgery compares favourably to international standards. Reconstruction of the complete vascular systems are carried out by experts in specialized centres. Sophisticated techniques are applied to the open and intravascular procedures of small and large vessels. Prosthesis grafts and vein autografts are often used with the help of atraumatic surgical equipment. The external quality control by the German Society of Vascular Surgery and the introduction of continuous medical education in vascular surgery help to maintain this high quality standard. There is a lack of vascular operation, a prerequisite for the development of further independent centres. PMID- 10666832 TI - [Cardiovascular risk in arterial vascular surgery reconstruction]. AB - Perioperative cardiac complications can limit the outcome after major vascular surgery where the underlying severe coronary artery disease is considered to be the main source. We describe the results of a prospective study including 201 patients undergoing elective vascular surgery. After looking at the encouraging low complication rate (mortality 0.99%, non-fatal myocardial infarction 2.98%, cardiac complications 9.95%) we would recommend the discussed diagnostic strategy. Specialized and expensive cardiac testing should be reserved for a few cases. PMID- 10666833 TI - [Promoting quality and quality circles from the viewpoint of established physicians--representative Bremen and Saxony-Anhalt results]. AB - RESEARCH QUESTION: To study the attitudes of ambulatory care physicians towards quality improvement, the intentions to join quality circles (QCs, peer review groups) and the expectations directed towards them. METHODS/SETTING: Survey with a five-page questionnaire posted to all ambulatory care physicians in the German states of Saxony-Anhalt (n = 3139) and Bremen (n = 1131). RESULTS: Response rates were 61.8% in Saxony-Anhalt and 41.7% in Bremen. 2412 questionnaires were available in this largest survey on that topic in Germany. Necessity of quality improvement (QI) in ambulatory care was approved by the majority of the respondents (1.7 on a 5-point Lickert scale). Concerns existed about a rise in control and the risk of abuse of QI measures. 56.4% in Saxony-Anhalt and 52.3% in Bremen had the intention to join a QC. Motives and impediments of participation in QCs were investigated by content analysis. A causal dominance analysis was performed to identify the key elements for the decision to participate. The main benefits of QC-participation were expected as assistance in daily practice and exchange of experiences. The major obstacles were professional and private duties, fear of control and inefficiency. CONCLUSIONS: Policies that could be adequate to rise motivations and tackle on widespread fears should be purposely adapted to the needs and expectations of the physicians. PMID- 10666834 TI - [Aspects of external quality assurance in anesthesiology--experiences in Hamburg]. AB - In 1994, external quality assurance in anaesthesia according to the German Society of Anaesthesiology and Intensive Care (DGAI) was obligatory introduced in Hamburg. Since 1992 in a pilot project and since 1994 compulsory nearly 500,000 anaesthesias were documented by 39 institutions with a standard data set issued by the DGAI and transferred to the project office of the Association for Quality Assurance (EQS) Hamburg. Comparative statistics of these data were produced at the project office. In the controlling committee and in meetings of the project participants the contents, policy and results of the project were critically analyzed and adjustments initiated whenever necessary. With an incidence of 14.1% of all anaesthesias with special occurrences (AVB), the results are in the same range compared to most other studies. To evaluate the concept of documentation the predictory power of single and combined risk assessments for the incidence of particular AVBs in elective anaesthesias were compared to the predictory power of ASA-Classification in order to reduce the parameters that had to be collected. This should lead to a positive influence on the quality of documentation. Besides one exception, no superior prediction power for AVB incidence could be demonstrated for any special risk assessment as compared with the ASA classification. This is also true for the AVBs which are associated with high lethality. Thus, the documentation of risk factors in the core data set as predictors can be abandoned without major loss of information. The participants consider the project to be a useful support for internal improvement projects. Besides the reduction of the amount of data in quality assurance to get a core of particularly meaningful parameters the classification of the surgical procedure by the ICPM- or OPS 301-Code should be integrated into the core data set of the DGAI. It would lead to an increase in acceptance of the method and thus to an increase in the validity of its results and valuations. PMID- 10666835 TI - [The "Hersbruck Model". Application and integration of the DIN EN ISO 9001 quality norms with criteria of the European Foundation for Quality Management in a clinic]. AB - The german health system has dramatically changed and still continues to do so. Modified aspects concerning economy, customer orientation, competition, quality assurance and quality management seem to be increasingly important. Appropriate response to this challenge demands a relevant adjustment of a quality "philosophy" within a hospital. The "Hersbruck Model" has proven to be a suitable approach: on the basis of a quality management system--established and certified according to DIN EN ISO 9001--it implements all components of the model of the European Foundation for Quality Management. The modern quality tools as Total Quality Management and continuous quality improvement allow a permanent increase of customer/patient satisfaction. PMID- 10666836 TI - 13C-methacetin breath test: isotope-selective nondispersive infrared spectrometry in comparison to isotope ratio mass spectrometry in volunteers and patients with liver cirrhosis. AB - The 13C-methacetin breath test (MBT) has been proposed for the noninvasive evaluation of the hepatic mixed function oxidase activity. Up to now, stable isotope analysis of carbon dioxide of the MBT has been carried out with isotope ratio mass spectrometry (IRMS). The aim of the present study was to test a recently developed isotope-selective nondispersive infrared spectrometer (NDIRS) in comparison to IRMS in healthy volunteers and patients with liver cirrhosis. Ten healthy volunteers (range 22 to 76 years) and ten patients with histologically proven liver cirrhosis (range 47 to 71 years; Child Pugh score A = 5, B = 3, C = 2) were studied. After an overnight fast each subject received 2 mg/kg BW of 13C-methacetin dissolved in 100 ml of tea. Breath samples were obtained before substrate administration and after 5, 10, 15, 20, 30, 40, 50, 60, 80, 100, 120, 150, 180 min. The 13C/12C-ratio was analyzed in each breath sample both by NDIRS (IRIS, Wagner Analysen Technik, Worpswede, Germany) and CF-IRMS (ABCA, Europa Scientific, Crewe, UK). Results were expressed as delta over baseline (DOB [/1000]) and as cumulative percentage doses of 13C recovered (cPDR [%]) at each time interval. Correlations between IRMS and NDIRS were tested by linear regression correlation. For measuring agreement an Altman-Bland-plot was performed. Applying correlation analysis a linear correlation was found (DOB: y = 1.068 +/- 0.0012.x + 2.088 +/- 0.2126, r = 0.98, p < 0.0001; cPDR: y = 1.148 +/- 0.0109.x + 0.569 +/- 0.172; r = 0.99, p < 0.0001). For DOB the mean difference (d) was 2.9/1000 and the standard deviation (SD) of the difference was 2.7/1000. The limits of agreement (d +/- SD) were -2.5/1000 and 8.3/1000. The comparison of DOB- and cPDR-values by NDIRS and IRMS shows a high linear correlation. However, the distance of the limits of agreement is wide. Consequently, the validity of the MBT could be influenced which could make MBT by NDIRS unprecise for exact evaluation of hepatocellular dysfunction. Further studies are necessary to determine sensitivity and specifity of the MBT with NDIRS in larger study populations. PMID- 10666837 TI - Evaluation of an enzyme immunoassay for detecting Helicobacter pylori antigens in human stool samples. AB - BACKGROUND AND AIM: So far, the detection of Helicobacter pylori (Hp) infection by stool analysis appeared to be almost impossible. With the Premier Platinum HpSA EIA a new enzyme immunoassay was developed for diagnosis of Hp infection, using polyclonal antibodies against Hp antigens in human stool. We evaluated this new test in its diagnostic accuracy in comparison to established reference methods. METHODS: From 54 consecutive patients (29 male, 25 female, age: 19 to 85 years) undergoing routine upper gastrointestinal endoscopy antral and corpus biopsies were taken for histology and Helicobacter urease test (HUT). Endoscopy, 13C-urea breath test (13C-UBT), serology, and stool probes sampling were performed within two days. Stool samples were aliquoted after reception and stored frozen (-20 degrees C) until tested. The Premier Platinum HpSA test (Meridian, Connecticut, Ohio, USA) was performed according to the manufactures protocol. Patients were considered to be infected with Hp if two of the four reference tests were positive. RESULTS: 28 of the 54 patients were Hp-infected. Only one of these was found to be false-negative by the HpSA EIA. Two false positive results were obtained in the noninfected group (sensitivity 96.4%, specificity 92.3%). CONCLUSION: In this group of patients investigated, the novel HpSA Enzyme Immunoassay (EIA) proved to be highly accurate for diagnosis of Hp infection. Collection and testing of stool are noninvasive and easy to perform, therefore this test will become an important tool for diagnosing Hp infection in clinical practice. PMID- 10666838 TI - [A new papillotome for cannulation, pre-cut or conventional papillotomy]. AB - A new papillotome was designed to overcome certain drawbacks of the needle-knife, that is most commonly used for precut sphincterotomies. The intention was to develop an instrument at least as good as the needle-knife or the Erlangen-type precut papillotome for precut procedures. In addition, it had to be suitable for direct cannulation of the biliary or pancreatic duct. PATIENTS AND METHODS: According to a prospective protocol 54 patients in whom a papillotomy was indicated were examined with the new instrument. The protocol allowed three futile attempts to cannulate or two inadvertant cannulations of the pancreatic duct with a standard cannula and hydrophilic guide wire before a precut was performed. The new baby-papillotome has a diameter of only 1 mm and a short 10 mm cutting wire. Similar to a guide wire it is introduced via a 6F- or 7F-introducer catheter. RESULTS: Cannulation of the desired duct (the bile duct in 48 patients, the pancreatic duct in five patients, Billroth II anatomy in three patients) was successful within one session in 98% (53/54). In one patient, the bile duct was successfully cannulated in a second session using the baby-papillotome, resulting in an overall success rate of 100%. Primary cannulation using the new papillotome without precut was obtained in 24% (13/54). Complications were mild pancreatitis in one patient and nonsignificant bleeding in three (immediate endoscopic hemostasis in all, no transfusions, no drop of hematocrit). There were no serious complications. CONCLUSION: The new baby-papillotome is suitable for precut as well as for primary cannulation. In this first series, the desired duct was cannulated in 98% within the first session with a low complication rate. Further studies of the new instrument seem desirable. PMID- 10666839 TI - [Prevalence of cholecystolithiasis in South Germany--an ultrasound study of 2,498 persons of a rural population]. AB - Gallbladder stones represent one of the most common reason for morbidity in western industrial nations. There remains a paucity of exact information regarding the prevalence and risk factors for this disease entity in Germany. As part of a whole-community survey focusing on the prevalence of echinococcosis multilocularis conducted in a population in southwestern Germany (response rate: 66.6%), 2,560 subjects underwent an upper abdominal ultrasound examination at which the presence of gallbladder stones was ascertained. In each case, upper abdominal sonography was performed following completion of a standardized interview. In 62 subjects, the gallbladder could not be adequately visualized due to an insufficient fasting period; the remaining 2,498 subjects (1,326 females, age 38.9 +/- 19.9 years; 1,172 males, age 37.7 +/- 18.8 years) were included in the study collective. Gallbladder stones (sonographically visualized gallbladder stones or history of cholecystectomy for cholecystolithiasis) were found in 196 participants (7.8%; 139 females [10.5%] versus 57 males [4.9%]). Statistical treatment of the data using multiple logistical regression techniques revealed a significant influence of the variables age, gender, body mass index (BMI) and positive family history on the development of gallbladder stones. The prevalence of gallbladder stones in the present study population is lower than figures reported for a study in Brandenburg and at 7.8% is rather low in comparison with other European studies. One explanation may be the low average age of study participants, almost 50% of whom were less than 35 years. Besides age, subjects' gender, BMI and positive family history were identified as significant risk factors. PMID- 10666840 TI - [Long-term collagenous sprue--remission with a gluten-free diet]. AB - We report on a 56-year-old caucasian female, 154 cm, 38 kg, who was admitted to the hospital in extremely poor, cachectic condition with pretibial edemas, ascites and pleural effusion with a history of severe watery diarrhea up to 15 times daily. The patient's history with permanent meteorism, recurrent episodes of watery diarrhea and dystrophy dates back to her early childhood. Multiple biopsies from the distal part of the duodenum showed a complete atrophy of villi with an extensive subepithelial layer of collagenous fibers. No antigliadin or antiendomysial antibodies could be detected. Under a strictly gluten-free diet, the patient's symptoms quickly improved. During the follow-up of one year, no recurrence of diarrhea and meteorism has been seen. The patient showed a continuous improvement of her general condition and nutritional state with a weight gain to 50 kg. Repeated control biopsies still revealed a pronounced villous atrophy. However, no subepithelial collagenous fibers were detectable. A diagnosis fitting to the patient's clinical symptoms of severe malabsorption, would be the rare "collagenous sprue", which is considered to be gluten insensitive. However, the patient's good response to a gluten-free diet raises the possibility, that the collagenous layer represents a manifestation of more severe forms of common celiac disease. In this case, it would not be necessary to regard collagenous sprue as a distinct entity. PMID- 10666841 TI - Autologous blood donation for surgery in inflammatory bowel disease--a report of six cases. AB - BACKGROUND: Surgery in inflammatory bowel disease (IBD) is frequently associated with need for perioperative blood transfusions carrying the potential risk of infection. Autologous blood donation is often limited by IBD-associated anemia which is reversible by intravenous iron and erythropoietin. We therefore tested the feasibility of autologous blood donation in IBD. METHODS: Six patients (five Crohn's disease, one ulcerative colitis) with indication for elective bowel resection were treated after informed consent was obtained. Two to four blood donations were scheduled during four weeks prior to surgery. Once a week 350-450 ml of blood were collected from patients with a hemoglobin level above 11.0 g/dl. After each donation 200 mg of iron saccharate diluted in 0.9% saline were given to all patients intravenously as substitute for donation-related iron loss. Patients with preexisting anemia or C-reactive protein above 2.0 mg/dl received concomitant erythropoietin. RESULTS: The scheduled number of packed red cells was donated successfully by four patients. Due to low hemoglobin levels two patients donated one unit less than intended. Four patients received autologous blood transfusions intra- or postoperatively. No patient needed homologous blood. No serious adverse events were observed during blood donations, perioperatively, and during the one year follow-up period. CONCLUSION: Preoperative autologous blood donation is save and feasible in IBD patients with elective bowel resection. PMID- 10666842 TI - [Treatment of multilocular hepatocellular carcinoma (HCC) of 4.5 cm and 3.5 cm diameter using percutaneous ethanol injection in a patient with advanced liver cirrhosis]. AB - Treatment of hepatocellular carcinoma (HCC) with percutaneous ethanol injection (PEI) is established for tumors up to 3 cm. We report on treatment of a multilocular HCC with a maximum size of 4.5 cm. CASE REPORT: In a 76-year-old woman with a liver cirrhosis (Child C) due to chronic hepatitis C HCC with two nodules (diameter 3.5 cm and 4.5 cm) was diagnosed. Because of the patient's reduced general state of health and the advanced cirrhosis surgical treatment and chemoembolization were declined. The two nodules were treated in ten settings during 15 weeks under ultrasound guidance with 85 ml of 96% ethanol. There were no severe side effects. The patient's general condition improved and serum alpha fetoprotein concentration decreased from 21,126 to 800 micrograms/l. Seven months after the diagnosis of the HCC she was admitted to another hospital due to a cerebral hemorrhage. A few days later she died because of a pneumonia. In spite of detailed micro- and macroscopically investigation no tumor was found during the autopsy. This case report shows that HCC up to a diameter of 4.5 cm can be effectively treated by PEI. This treatment modality is cheap and well-tolerated even in patients suffering from advanced cirrhosis. PMID- 10666843 TI - [Hereditary hemochromatosis--new developments after discovery of the HFE gene]. AB - Hereditary hemochromatosis (HH) is an autosomal recessive disorder of iron metabolism, resulting in an increased iron deposition and multiorgan failure. Recently a candidate gene of HH, termed HFE, has been identified on chromosome 6, coding for a protein homologous to major histocompatibility complex (MHC) class I molecules. Two mutations of the hemochromatosis gene leading to an exchange of cysteine to tyrosine at aminoacid 282 and histidine to asparagine at aminoacid 63, are retained responsible for the development of hereditary hemochromatosis. The Cys282Tyr-mutation disrupts a disulfid bond and thus abrogates binding of the mutant HFE-protein to beta 2-microglobulin and its presentation on the cell surface. The His63Asp-mutation seems to play a role in pH-regulated dissociation of the transferrin receptor/transferrin complex in the lysosome. Mutations of the HFE-protein alter the affinity of the transferrin receptor for its ligand transferrin and may thus cause an intracellular accumulation of iron. Knowledge of the responsible gene allows a molecular diagnosis of HH. The new genetic marker can be used for screening and confirmation of HH reducing the need for confirmatory liver biopsies. Compared to standard screening parameters like ferritin and transferrin saturation genetic testing will allow the diagnosis of HH in an early, asymptomatic state before iron accumulation has occurred. As a normal life expectancy of patients with HH can be achieved if iron reduction is initiated early, genetic testing may thus be of great benefit for patients with HH. PMID- 10666844 TI - [Gene therapy of pancreatic carcinoma]. PMID- 10666845 TI - [Incidence and prognosis of collagenous and lymphocytic colitis]. PMID- 10666846 TI - [Identification of CD81 protein as HCV receptor--relevance for pathogenesis and cell tropism?]. PMID- 10666847 TI - [Evidence-based evaluation of the back school. Wide distribution with minimal effectiveness?]. PMID- 10666848 TI - [Tissue engineering of bone tissue]. PMID- 10666849 TI - [Torsion changes in the throwing arm of handball players]. PMID- 10666850 TI - [Orthopedics and geriatrics]. PMID- 10666851 TI - [Incidence of joint-specific risk factors in patients with advanced cox- and gonarthroses in the Ulm Osteoarthrosis Study]. AB - AIM OF STUDY: To determine the prevalence of joint specific risk factors in patients with different patterns of advanced hip and knee osteoarthritis (OA). METHODS: We performed a cross-sectional multicenter study in four orthopaedic hospitals in the southwest of Germany. A detailed medical history (date and nature of trauma, conservative and surgical treatment of congenital or acquired joint disorders known as secondary causes of OA) and radiographic evaluation (sequelae of hip dysplasia, slipped capital femoral epiphysis or other malformations) was obtained in 809 patients with advanced hip (n = 420) or knee (n = 389) osteoarthritis, which required unilateral total joint replacement. According to the presence or absence of joint specific risk factors, patients were classified as having secondary or primary (idiopathic) OA. RESULTS: In 41.7% (25.5%) of patients with hip OA and 33.4% (11.1%) of patients with knee OA some predisposing abnormality of the operated (or contralateral) joint could be observed. In hip OA the underlying pathological conditions were mainly hip dysplasia (25.0% in the operated joint and 14.8% in the non-operated joint) and slipped capital femoral epiphysis (7.1% and 14.8%), while knee OA was most often associated with a history of severe trauma (28.6% and 8.3%) CONCLUSION: While there is a lack of comparable investigations in patients with advanced knee OA, the presented data is somewhat contradictory to earlier reports of the prevalence of identified underlying risk factors in patients with hip OA. The reported differences, however, might be attributed to different methodological approaches and could also resemble recent changes in the multifactorial ethiopathologic concept of OA. PMID- 10666852 TI - [Diagnosis and therapy of spinal stenosis in the elderly]. AB - PROBLEM: Spinal stenosis is a common and increasing problem in the elderly population and a diagnostic and therapeutic challenge. METHODS: An overview of etiology, epidemiology, diagnostics and therapy is given based on a literature review of the years 1978-1998 and experiences since 1985. RESULTS: Dealing with spinal stenosis we have to distinguish concerning diagnostics and therapy between cervical and lumbar spinal stenosis. The cardinal symptom of the lumbar spinal stenosis is the claudicatio spinalis and of the cervical spinal stenosis the cervical myelopathy occasionally combined with radiculopathy. The first therapeutic step should be the conservative therapy in nearly all cases, the only exception is a severe cervical myelopathy where an operation is indicated. In case of persistent or progressive symptoms under a sufficient conservative therapy, operative therapy is indicated. The different possible therapy decisions will be based on flow-charts. CONCLUSION: Whereas in lumbar spinal stenosis the indication for operative therapy should be considered with reservation, in cervical spinal stenosis with myelopathy operative therapy should be considered at an early stage. PMID- 10666853 TI - [Epidemiology of accidental falls in the elderly]. AB - INTRODUCTION: There is a lack of epidemiological data on falls and fall-related injuries for the aged population in Germany. The purpose of the article is to present the available data focusing on the incidence of the fracture types that carry the highest risk for mortality, hospitalization and persisting disabilities. METHODS: The paper reports on a 10% representative sample that was drawn by the Federal Bureau of Statistics. A medline search for 1980-1998 was performed to identify relevant articles. The OECD database was used for mortality rates after injurious falls. If no sufficient data were published for Germany, incidence rates of demographic comparable European regions are reported. RESULTS: The number of fractures of the lower extremity (ICD 820-829) followed by hospitalization was 139,000 in 1996 for elderly (65+ years). Fractures of the upper extremities (ICD 810-819) requiring hospitalisation were reported for 65,000 persons aged 65+ years and head trauma (ICD 850-851) followed by hospitalization in this age group for 29,000 persons. There remains an information gap on the incidence of falls and fall-related sequelae without hospitalization in Germany. CONSEQUENCES: Facing, the demographic transgression the prevention of falls and fall-related injuries gains a high priority. Operative management and rehabilitation procedures for elderly trauma patients should be further evaluated and improved. PMID- 10666854 TI - [Fractures of the upper extremity in the elderly]. AB - PROBLEM: The primary goal in treating fractures in elderly persons is safe and rapid restoration of their functional capabilities to secure independence in everyday activities. The intention of this paper is to present an overview of treatment principles useful in this age group. METHODS: We analysed a series of 404 patients above the age of 70 years who underwent surgery for fractures of the arm between 1981 and 1997. RESULTS: Diaphyseal fractures are less frequent in this age group and their treatment does not differ significantly from principles established for younger patients. The majority of fractures of the proximal humerus and the distal radius can be treated by conservative means. Unstable fractures are frequent in the distal humerus and the olecranon requiring operative reduction and osteosynthesis. CONCLUSIONS: Preferred operative techniques in aged persons inflict minimal surgical trauma and have a low rate of complications and secondary interventions. In contrast, anatomical reconstruction has the highest priority in younger individuals. PMID- 10666855 TI - [Therapy of para-articular hip fractures in the elderly]. AB - Fractures of the hip in old patients are life-threatening events. A steady increase of that fracture type is likely in the near future. Surgical therapies and strategies have to consider the special requirements and problems of geriatric patients to achieve better results. They have to be designed together with new concepts of geriatric rehabilitation programmes. MATERIAL AND METHODS: In 1992 148 patients with hip fractures were treated at the department of traumatology, University Ulm. Follow-up parameters were daily activities in life pre- and postoperatively as well as mortality, type of fracture and surgical treatment. RESULTS: There were 79 femur fractures of the collum and 69 fractures of the trochanteric region. Mean age was at 81.2 and 81.9 years respectively (68 f/11 m versus 52 f/13 m). At the time point of accident 84 patients lived at home whereas the remaining stayed at a nursing home. The highest mortality was found in patients living in a nursing home (93%). The overall mortality one year following the trauma was 26%. 50% of the patients had hip prosthetic replacement, the remaining received a gliding screw, 9 patients a proximal femoral nail or lag screws (n = 6). CONCLUSION: Our results demonstrate that hip fractures in geriatric patients have a high mortality, especially in those living in a nursing home. The surgical concepts should aim to reduce that number and to allow the same daily activity of life as preoperatively. The main part in these concepts is an early start of geriatric rehabilitation. There are at least two groups. On the one hand, the active old patient who acquires his fracture during an activity. In this cases the aim must be the full rehabilitation and afterwards returning to normal environment. On the other hand there are the patients living in a nursing home who have the highest risk of injury related death. In these patients the first aim must be prevention of the accident. PMID- 10666856 TI - [Thoracoscopic interventions in deformities of the thoracic spine]. AB - AIM OF THE STUDY: We prospectively studied 9 patients with deformities of the thoracic spine who underwent thoracoscopic surgery to critically evaluate the benefits and limitations of thoracoscopy. METHODS: Seven patients with deformities of the thoracic spine (5 scoliosis, 2 kyphosis) underwent a thoracoscopic release and posterior correction and fusion in a single stage. In one case of a crankshaft-phenomenon a thoracoscopic epiphyseodesis und in another case of a posttraumatic kyphosis a thoracoscopic instrumentation and fusion were performed. The average age was 21 years, the follow-up was 18 months with a minimum of 12 months. The perioperative data including complications were collected and a radiographic analysis concerning curve correction was carried out. RESULTS: The scoliotic curves measured preoperatively 84 degrees on average with a Cobb angle of 62 degrees on the traction films and were corrected by 57% to averagely 36 degrees at follow-up. In the two cases of Scheuermann kyphosis a preoperative kyphosis of 94 degrees respectively 82 degrees was corrected to 52 degrees respectively 58 degrees. Between 4 and 5 discs were excised with an average operative time of 160 min and a blood loss of 380 ml. A conversion to open thoracotomy was not necessary in any case. There were no intraoperative neurovascular complications. CONCLUSIONS: Thoracoscopic procedures in deformities of the thoracic spine are technically demanding; however, it is a minimally invasive procedure with a reduced approach-related morbidity compared to open thoracotomy. The indications for a thoracoscopic release are rigid kyphosis and scoliosis with rigid curves between 80 and 90 degrees Cobb angle in which an anterior correction and instrumentation alone is not considered. PMID- 10666857 TI - [Guillain-Barre syndrome as differential diagnosis of intervertebral disk-induced nerve root compression]. AB - INTRODUCTION: Diagnosis of Guillian-Barre Syndrome usually is not difficult, but diagnostic failure occurs for the variable initial presentation. Diagnosis is based on physical examination showing loss of motor strength in more than one limb and loss of deep tendon reflexes. Ventilatory assistance, pharmacologic maintenance of cardiovascular homeostasis, corticosteroids, IgG and plasma exchange are the dominant therapeutic measures. CASE: This article reports on a case of a 59-year old surgeon suffering from degenerative disc disease in the lumbar spine. The patient developed a severe course of the Guillian-Barre Syndrome with persisting motor weakness of the legs. CONCLUSION: If the primary problem at presentation is limb and back pain the pathology appears to be in the musculoskeletal rather than in neurological system. The awareness of this presentation of Guillian-Barre-Syndrome will eliminate delay in diagnosis. PMID- 10666858 TI - [Early diagnosis of isthmic spondylolysis with MRI]. AB - INTRODUCTION: Early diagnosis of isthmic lumbar spondylolysis cannot always be established on plain radiographs and CT scans, only. In the case presented here, magnetic resonance imaging (MRI) showed typical bone marrow changes in T1- and T2 weighted images, even at an early stage. CASE: A 11-year old female judoka complained of deep lumbar pain with local tenderness to pressure at L3 to S1. Clinically, there was no neurologic deficit. Conventional x-ray showed no abnormalities. In contrast, MRI revealed a locally ill-defined bone marrow oedema in both pars interarticularis of the 5th lumbar vertebra. This was interpreted as the typical MR-tomographic feature of occult stress fracture, which has to be seen as early evidence of isthmic spondylolysis. Complete restitution was achieved after conservative treatment. CONCLUSION: In early spondylolysis- presented here in form of a case report--, changes of MR signal intensity in the pars interarticularis may be detected, even before fracture lines are to be seen on plain radiographs. Further studies are necessary to confirm MRI to be the method of choice for early diagnosis. PMID- 10666859 TI - [Mannerfelt arthrodesis of the wrist joint in patients with chronic polyarthritis. A retrospective analysis of 24 cases]. AB - AIM: Mannerfelt established his technique of wrist arthrodesis with stabilisation by an intraosseous rushpin as a secure method for patients with rheumatoid arthritis. This study was performed to evaluate the mid-term results in a consecutive group of patients. METHODS: Out of a group of 39 operations 24 wrist arthrodeses (61%) in 19 patients have been followed 12-96 months postoperatively (average 44 mths) by clinical testing and radiographic examination. All operations were performed in the original technique. All patients suffered from rheumatoid arthritis in an advanced stage (Larsen III-V). RESULTS: All but one patient were free of pain. Function and strength of the hand increased significantly in all patients. All patients had additional resection of the ulnar head that led to normal pro-supination of the forearm. 18 patients were very satisfied with the result of the procedure. All but one of the wrists showed complete fusion. In one case there was an intraoperative perforation of the pin through the radial cortex, in another case we saw a fissure of the shaft of the third metacarpal bone. One patient showed a dysesthesia in the third finger. CONCLUSION: The results in this group of patients confirmed the advantages of Mannerfelt's technique such as simple operative technique, high fusion rate and low incidence of complications. PMID- 10666860 TI - [Functional and socioeconomic outcome of inpatient rehabilitation of patients with complex hand injuries]. AB - PURPOSE: Evaluation of functional outcome and socio-economic results of patients with complex hand injuries after inpatient rehabilitation. PATIENTS AND METHODS: In a prospective investigation 36 patients with complex hand injuries received an intensive therapy regime under inpatient conditions. In all patients functional parameters of the hand (total active range of motion, palm to palm distance, spread between thumb and index finger, spread between thumb and little finger) and the score of Buck-Gramcko were registered initially and at the end of inpatient rehabilitation. Socio-economic data (kind of discharge, job reintegration, further treatment) were documented as well. RESULTS: The total active range of motion could be improved from 36% to 69%, spread D1-D5 from 18.5 to 20.7 cm and spread D1-D2 from 13.9 to 15.6 cm. The palp to palm distance of all fingers was reduced from 4.6 cm to 2.3 cm. The Buck-Gramcko score of all injured fingers improved from 4.2 points (poor) at admission, to 11.8 points (good) at discharge, At the end of inpatient rehabilitation 18 patients could return to their job either gradually or on full-time. CONCLUSION: Even patients with complex hand injuries, good functional results could be achieved by an intensive inpatient rehabilitation characterised by close medical supervision and guidance and tight clinical controls. PMID- 10666861 TI - [Hallux valgus: a therapy concept and its outcome from 1993 to 1996]. AB - INTRODUCTION: Aim of this study is to present the treatment concept and the results of hallux valgus surgery of our department. PATIENTS AND METHODS: The criteria for decision making are 1st intermetatarsal angle, congruency of the first metatarsophalangeal joint and sesamoid position. According to our concept we performed between 1993 and 1996 42 Chevron osteotomies for mild, 138 Chevron with lateral soft tissue release (Chevron + LSR) for moderate and 93 basal crescentic (Mann) osteotomies with lateral soft tissue release for severe hallux valgus deformities. RESULTS: 31 Chevron, 118 Chevron with lateral soft tissue release and 80 basal crescentic osteotomies (Mann) were seen at an average follow up of 19 months (12-29) after a Chevron, respectively. 16 months (12-43) after Chevron with lateral soft tissue release and 18 months (12-32) after a basal crescentic osteotomy. 83% of all patients classified the results of the surgery as "very satisfactory" and "satisfactory". The average hallux valgus angle was corrected from preoperatively. 25.3 degrees (Chevron), 29.9 degrees (Chevron + LSR) and 41.8 degrees (Mann) to 16.5 degrees (Chevron), 12.1 degrees (Chevron + LSR) and 14.1 degrees (Mann) at final follow up, the average first intermetatarsal angle was corrected from preoperatively 12.1 degrees (Chevron), 14.0 degrees (Chevron + LSR) and 17.4 degrees (Mann) to 7.9 degrees (Chevron), 5.8 degrees (Chevron + LSR) and 7.8 degrees (Mann) at final follow up. CONCLUSION: Our analysis of the three osteotomies revealed that with this differentiated concept we were able to achieve excellent and good results in more than 80% of our patients. Only with a treatment plan that includes different procedures to address the various stages of hallux valgus can one achieve the optimum result for the patient. PMID- 10666862 TI - [Osteogenesis in exposure to ionizing radiation in vitro]. AB - Irradiation is a well established therapeutical concept to prevent heterotopic ossification after joint replacement. The influence of irradiation on proliferation of mature osteoblasts and their potential osteoprogenitors, matrix formation and mineralization are not well known in this setting. We therefore studied the effect of different doses of ionizing irradiation on the several steps of osteogenesis in vitro, using cells isolated from the juvenile rat. A colony forming test, the MTT-viability assay, a cell count, measurement of the cellular protein content and alkaline phosphatase activity, as well as determination of in vitro mineralisation have been applied to calvarian osteoblasts, fibroblasts and stromal bone marrow cells. Irradiation results in a dose-dependent suppression of clonogenic activity in all mitotically active cells, but metabolic activity and matrix synthesis were not impaired. In dense cultures alkaline phosphatase expression and in vitro mineralisation were not significantly affected by irradiation. Our experimental in vitro data suggest that irradiation inhibits the initial phase of in vivo osteogenesis due to the cytostatic effect. Postoperative irradiation after THR must therefore take place as early as possible. The homoeostasis of normal, orthotopic bone does not seem to be severely affected by local low-dose irradiation. PMID- 10666863 TI - [Heparin-induced thrombocytopenia after elective hip joint replacement with postoperative prevention of thromboembolism with low-molecular-weight heparin]. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is a severe side effect of the prophylaxis of venous thromboembolism with unfractionated heparin. The aim of the present study is to gain more information on the incidence of HIT during prophylaxis of venous thromboembolism with low-molecular-weight heparin in elective hip surgery. METHODS: 586 consecutive patients were included into the prospective study, who were admitted to hospital for elective hip replacement. The incidence of thrombocytopenia, clinically manifest venous thromboembolism and of the heparin-induced IgG antibodies were analysed during prophylaxis with low molecular-weight heparin. Patients received once daily subcutaneously low molecular-weight heparin for a mean of 28 days postoperatively. Platelet counts and clinical examinations for the presence of venous thromboembolism were done at days 0, 2, 7 (+/- 1) and 12 (+/- 2). Heparin-induced IgG antibodies were determined before and after a 12 (+/- 2) days prophylaxis with low molecular weight heparin in 265 of 586 patients randomly. Patients were reexamined for thromboembolic complications after 3 and 6 months. The clinical suspicion of thromboembolic complication was documented objectively. RESULTS: None of the patients developed a decrease of platelets of < 50% of the initial value. Ten of 265 patients had elevated IgG antibodies against heparin/platelet factor 4 before prophylaxis (3.8%). After the 12 (+/- 2) days prophylaxis 13 of 265 patients had elevated IgG antibodies (4.9%). C14 serotonin assay was positive in 0 of 10 patients before treatment and in 3 of 19 patients at day 12 (+/- 2). Ten patients developed venous thromboembolism postoperatively (8 x deep venous thrombosis, 2 x pulmonary embolism, no fatal embolism). Only 1/19 patients with elevated antiheparin IgG titres developed venous thromboembolism. The C14 serotonin assay was negative in this patient. Two patients died in the postoperative phase due to underlying cardiovascular diseases. CONCLUSIONS: In patients with elective hip replacement prophylaxis of venous thromboembolism with low molecular-weight heparin was associated with a very low incidence of HIT, and hence screening for HIT antibodies is not required. PMID- 10666864 TI - [Collagen type VI content in healthy and arthritis knee joint cartilage]. AB - AIM OF THE STUDY: Since collagen type VI seems to play an important role in cartilage metabolism and is increased in osteoarthritis, the aim of this study was to investigate whether different stages of osteoarthritis can be characterised by the content of collagen type VI in human knee cartilage. MATERIAL AND METHODS: Collagen type VI was investigated in 148 histologically normal and 117 osteoarthritic cartilage samples from 18 different localisations of human knee joints. It was quantified in cartilage extracts using an inhibition ELISA. RESULTS: In normal cartilage the average content of collagen type VI was 0.48 per cent of total collagens. Interestingly, there was a high variability not only in osteoarthritic, but also in normal cartilage. The statistical analysis showed significant differences between normal femoral, tibial or retropatellar cartilage samples. Therefore, normal and osteoarthritic samples from different localizations had to be compared separately. A significant increase of collagen type VI was already found in early osteoarthritic lesions. CONCLUSIONS: Inspite of a statistically significant increase of collagen type VI in osteoarthritic cartilage, the range of concentrations found in normal and osteoarthritic samples overlap. In view of the high interindividual variability, collagen type VI is not very precise in the diagnosis of early osteoarthritic lesions if used as the only marker. PMID- 10666865 TI - [Hemostaseology and pre-eclampsia]. PMID- 10666866 TI - [Pre-eclampsia--endothelial damage of endothelial activation?]. AB - There is considerable evidence that endothelial damage, followed by the release of vasoactive substances contributes to the pathophysiology of preeclampsia. Because of controversial experiences in literature we wanted to evaluate the potential cytotoxic effect of preeclamptic sera on cultured endothelial cells. Therefore cultured human umbilical vein endothelial cells (HUVEC) were stimulated with sera obtained from preeclamptic patients, while sera from normotensive pregnant and nonpregnant women served as controls. To prove the viability of these cells we performed ethidiumbromide/acridinorange immunostaining and determined t-PA/PAI-1 release into the supernatant. These experiments could not show any cytotoxic effect on endothelial cells. In ongoing studies we measured the concentrations of adhesion molecules, markers of endothelial activation, in maternal sera, in the supernatant of cultured endothelial cells, and on cell surface after stimulation with the above mentioned sera. In the supernatant we couldn't determine any different concentrations of adhesion molecules after stimulation with the different sera, but using immunofluorescence-microscopy an increased concentration of those molecules could be detected on the endothelial surface after stimulation with preeclamptic sera than compared to sera from normotensive controls. In conclusion, our experiments support the hypothesis that sera from preeclamptic women may cause endothelial activation. PMID- 10666867 TI - [Thrombocyte activation in pre-eclampsia]. PMID- 10666868 TI - [Trophoblast invasion in pre-eclampsia]. AB - It is generally accepted that insufficient invasion of trophoblast cells into the myometrial portions of the spiral arteries is thought to play a crucial role in the development of preeclampsia. As a consequence, uteroplacental vessels fail to undergo adaptive changes which are imperative to provide a sufficient blood supply to the placenta. Consecutive placental hypoxia is supposed to cause secretion/shedding of still unidentified placental metabolites resulting in different forms of pregnancy-induced hypertension. This review presents published data concerning the causes of insufficient trophoblast invasion in preeclampsia. Expression of HLA-G by extravillous trophoblast cells seems to be altered, resulting in activation of the maternal immune system. The pattern of integrin expression as well as the secretion of proteases is reported to be disturbed, which could lead to a reduced invasive potential of the trophoblast cells. Recent data indicate a pathophysiological role of NK-cells and macrophages in the altered trophoblast invasion. Also angiotensinogen Thr235 polymorphism seems to alter early physiologic changes in spiral arteries. In summary, preeclampsia seems to be induced by a multifactorial disturbance of trophoblast invasiveness which is characterized by reduced invasiveness of the trophoblast cells themselves and by an activated maternal immune response blocking the invasion by the semiallogenic trophoblast. PMID- 10666869 TI - [Animal experiment models of hypertension in pregnancy--an alternative to clinical studies?]. AB - Because of ethical constraints upon research during human pregnancy and of the heterogeneity of preeclampsia a number of experimental animal models were used to study pathophysiology of preeclampsia. We reduced the uteroplacental flow by aortic clip technique in 72 SHR-rats in 4 groups (G 1 not gravid--no clip, G 2 not gravid--aortic clip, G 3 gravid--no clip, G 4 gravid--aortic clip). While gravidity normalised blood pressure in SHR (G 3), reduction of the uteroplacental flow significantly increased maternal blood pressure (G1 240/180 mmHG, G2 238/183 mmHg, G3 148/99 mmHG, G4 200/145 mmHg) and leads to significant higher hematocrit, significant lower plasma renin activity and aldosterone plasma concentration, as it is found in human superimposed preeclampsia. SDS-page electrophoresis proved a significant increase of urinary high density proteins. CONCLUSION: In comparison to chronical inhibition of NO-synthesis or chronical application of ultra-low-doses-endotoxin, reduction of uterine flow by aortic compression is a reliable animal model and leads in SHR to a superimposed preeclampsia-like syndrome with many similarities to human clinical findings. Animal models for preeclampsia are no alternative to clinical studies, but they are furthering our understanding of pathophysiology in many fields. PMID- 10666870 TI - [Significance of the renin-angiotensin-aldosterone system for pathogenesis and early diagnosis of pregnancy-induced hypertension with special reference to mineralocorticoid receptors]. AB - OBJECTIVE: In women with pregnancy-induced hypertension (PIH) the density of mineralocorticoid receptors (MR) in human mononuclear leukocytes (HML) is reduced compared with healthy pregnant women. The same applies to plasma levels of aldosterone and 18-hydroxycorticosterone. In this study, we investigated whether alterations of these parameters preceded the development of clinical symptoms and, therefore, might be potential predictors of PIH involved in the pathogenesis. PATIENTS AND METHODS: In eighty-four women belonging to the risk group for PIH but not showing any symptoms neither of PIH nor preeclampsia (PE) we characterized prospectively before the onset of disease in the second trimester of pregnancy mineralocorticoid receptor status in HML and steroid plasma levels of aldosterone and its precursors as well as cortisol through radioimmunoassay. RESULTS: 15 women developed PIH, three of which developed PE. Neither in the density of MR nor in the affinity the women that developed PIH showed any difference from healthy women. Steroid plasma levels were identical as well. CONCLUSION: We conclude that a reduction of mineralocorticoid receptors does not precede PIH within the peripheral blood. But still one can assume that the RAAS may be involved in the pathogenesis of PIH, possibly on a local level within the placenta or as a secondary change, initiated by still unknown factors. PMID- 10666871 TI - [The autonomic nervous system and pre-eclampsia]. AB - Alterations in the autonomic cardiovascular control have been implicated to play an important etiologic role in preeclampsia. Earlier assessments of sympathetic nervous system activity by measurements of urinary and plasma levels of catecholamines showed contradictory results, due to serious methodological problems. The microneurographic technique enables the assessment of sympathetic outflow to the vascular bed of skeletal muscles. By the use of this technique it has been shown that preeclampsia represents a state of sympathetic overactivity as compared to healthy pregnant women. Other studies have shown that exogenous stimulation of the celiac ganglion causes a HELLP-syndrome-like disease in pregnant rats and that preeclamptic human plasma increases norepinephrine release in isolated sympathetic neurons of chicken embryos. These data suggest that the autonomic nervous system may play a fundamental role in the etiology of preeclampsia. PMID- 10666872 TI - [Hypertensive illnesses in pregnancy: when is ambulatory management possible, when is hospitalization indicated?]. AB - Hypertensive disorders complicating pregnancy are the most common medical complications of pregnancy and are a major cause of maternal and perinatal morbidity and mortality. Thorough ambulatory obstetric care is likely to achieve a risk reduction. The main topics of ambulatory obstetric care are early identification of typical signs of preeclampsia, detection of uteroplacental insufficiency, and their consequences, detailed information of the patient, and early admission to a specialised obstetric care unit or perinatal center. Early diagnosis, close medical supervision, and timely delivery are the keys of the treatment of preeclampsia. PMID- 10666873 TI - [Doppler ultrasound findings in therapy with urapidil]. AB - OBJECTIVE: In a prospective study we investigated, whether changes in fetal or maternal circulation can be found by Doppler sonography under therapy with urapidil. PATIENTS AND METHODS: We investigated in 11 patients with severe pregnancy-induced hypertension (PIH) or superimposed preeclampsia in the third trimester. Doppler flow results of maternal and fetal vessels before and during therapy with urapidil. Hemodynamic parameters, like maternal blood pressure, and heart rate as well as other clinical parameters were assessed. RESULTS: Whereas the antihypertensive effects in the systolic and diastolic blood pressure were significant, we couldn't find changes in the resistance indices of fetal circulation. In contrast we found a decrease in the resistance indices in maternal vessels after an average of six days of urapidil therapy. These changes were significant in the uterine artery on the side of placental insertion. CONCLUSIONS: Because of only few side effects and a safe lowering of blood pressure, therapy with urapidil seems preferable to other well known drugs. Although Doppler flow velocimetry of uteroplacental vessels has no diagnostic benefit in monitoring PIH, the findings with this method under prolonged antihypertensive therapy are a chance for a better understanding of hemodynamic changes. The decrease in the resistance indices is another argument for the use of urapidil. PMID- 10666874 TI - [Early detection of pre-eclampsia within the scope of prenatal care]. AB - Pregnancy-induced hypertension and preeclampsia account for about 25% of perinatal morbidity and mortality and are a leading cause of maternal death in developed countries. Consequently, early diagnosis of the disease seems very important. Numerous clinical and biochemical methods have been described and tested for the prediction of preeclampsia. The results of these studies are critically described here. In summary, no test presently available serve as a reliable early marker of developing preeclampsia. However, vaginal Dopplerultrasound may allow prediction as early as the 12th to 16th week of gestation. Moreover, methods detecting underlying mechanisms of the disease such as endothelial dysfunction and platelet activation, are being developed and might provide improved early identification of pregnant women at high risk. PMID- 10666875 TI - [The role of antiphospholipid antibodies in patients with pre-eclampsia]. PMID- 10666876 TI - [Pre-eclampsia and anesthesia]. AB - The management of anaesthesia in patients suffering preeclampsia has to be selected individually. There is a high rate of caesarean sections in patients with preeclampsia. Intubation anaesthesia or regional anaesthesia are commonly used methods and can be considered comparable and equally useful. In our opinion, the application of regional anaesthesia should be preferred, if the initial criteria, such as normal neurologic status and blood coagulation, are fulfilled. We believe in a considerable reduction of vital risks when using regional anaesthesia. Our aim was to figure out the risk-factors of preeclampsia and preterm maturity regarding anaesthesia management. Preeclampsia is a severe illness in pregnancy with a frequency of 5-10%. In 1997, we experienced 73 patients with praeeclampsia according to the definition of ISSPH. Mode of delivery was conventional in 10 patients, 2 patients had a vacuum extraction and 61 patients needed a caesarean section. The rate of epidural anesthesia for delivery was 51%. We used epidural anaesthesia in 6 spontaneous deliveries, in 2 vacuum extractions and in 30 caesarean sections. There were no problems with epidural anaesthesia and the outcome of mother and child was also considered to be excellent. We suggest to use regional anesthesia techniques whenever possible. PMID- 10666877 TI - [Current aspects of antihypertensive therapy in pregnant patients with pre eclampsia]. AB - Preeclampsia is a disease which occurs in Europe in about 6-8%, in the USA in about 7-10% and in Africa in about 18% of all pregnancies. A causal treatment of preeclampsia is, with the exception of delivery, not possible up to now. Since a prematurely delivery of the newborn has to be avoided because of the risks caused by immaturity of lungs, treatment and care of pregnant women having preeclampsia or any other kind of hypertensive diseases is restricted to the following approaches: antihypertensive treatment, volume expansion, and eclampsia prophylaxis with magnesium sulfate. Object of this treatment is to avoid complications on the mother's side caused by the disease and to postpone delivery, as far as possible from the child's side, in order to reduce the consequences of premature birth. During antihypertensive treatment of patients with serious hypertension, i.e. with diastolic blood pressure of 110 x mm Hg and higher, dihydralazine is in clinical use since 40 years, although many patients suffer from side-effects of dihydralazine such as distinctive tachycardia, headaches, fluid retention and nausea. With urapidil a well controllable antihypertensive is available, which prevents the effect of catecholamines at the vascular wall by a postsynaptic alpha-1 receptor blockade. Previous studies related to the application of urapidil in the treatment of hypertension during pregnancy certify the good controllability of urapidil following intravenous application as well as minor side-effects after start of treatment. PMID- 10666878 TI - [Epidemiology, risk factors and predictors of pre-eclampsia]. PMID- 10666879 TI - [Psychiatry in XXI century]. PMID- 10666880 TI - [Cerebral strokes at young age]. AB - 791 patients aged 15-44 years with different forms of cerebral stroke that accounted for 9.4% from all the patients hospitalized because of stroke were treated. Ischemic stroke (IS) was diagnosed in 477 patients (60.3%), hemorrhagic stroke (HS) in 293 patients (37.3%), thromboses of the sinuses and veins of the brain in 19 patients (2.4%). IS to HS was 1.6:1; cerebral strokes were observed in men twice as frequently as in women. The main causes of HS (180 men, 115 women) were anomalies of cerebral vessels and arterial hypertension. Intracerebral and subarachnoidal hemorrhage occured with the same frequency. 38% of the patients died. The main causes of IS in 477 patients (285 men, 192 women) were arterial hypertension, rheumatism and atherosclerosis of cerebral and precerebral arteries. Embolic strokes occurred 4,5 times more frequently in women, than in men. 6.7% of the patients died. Among the patients with disorders of venous cerebral circulation (13 women, 6 men) 2 women with thrombosis of upper longitidinal sinus died. According to authors' data pregnancy and delivery are a significant risk factor for development of all forms of cerebral stroke. PMID- 10666881 TI - [Dreams and psychologic defence in children and juveniles with neurotic disorders]. AB - The examination of 78 patients at different stages of neurotic disorders and 25 healthy individuals (10-17 years old) has shown that the dynamics of dreams at compensatory stage was directed to their intensification, affective and cognitive activation and to reduction of dreams' activity during decompensation of neurotic disorders. This may indicate their role in psychologic defence. In neurotic patients peculiarity of the reaction to stress was the presence of some latent period after stress, that preceded the very reaction of dream to stress, its longer duration than in healthy individuals and its qualitative peculiarity. That may reflect complexity of adaptation to stress influence in neurotic patients. PMID- 10666882 TI - [Clinical features of ischemic heart disease and modes of subjective perception of illness]. AB - The sample included 91 inpatients with different clinical forms of ischemic heart disease (IHD) and modes of subjective perception of illness (subjective meaning of illness). Prichard's Reaction to Illness Questionnaire, Hospital Anxiety and Depressive Scale, Rotter's Internal-External Control Scale and Illness Locus of Control Scale (Bevz I.A.,1998) were used on day 14 after admission for qualification of the patient's subjective perception of illness. The following clinical predictors of hypernosognia (inadequately high subjective significance of illness) were revealed: 1) the onset of IHD in midlife (<65 years) with its subsequent fast progression including high incidence of recurrent coronary events and/or congestive heart failure, 2) "typical" and protracted angina pectoris, 3) cardiac arrhythmias accompanying persistent high heart rate (sinus tachycardia, chronic atrial fibrillation, frequent extrasystoles) and defying any self-care, and 4) severe heart failure. On the other hand clinical predictors of hyponosognosia (inadequately low subjective significance of illness) included 1) the onset of IHD in elderly individuals (>65 years) and its subsequent slow progression without recurrent coronary events and/or congestive heart failure, 2) the socalled "anginal syndrome" (lack of angina's coupling with psychical exertion, atypical pain location, inconstant efficiency of nitroglycerin) and silent myocardial ischemia, 3) the paroxysmal cardiac arrhythmias (infrequent extrasystoles, paroxysmal atrial fibrillation, supraventricular tachyarrhythmias) with normal or slow heart rate between the paroxysms and high efficiency of self care, and 4) mild to moderate heart failure. The findings are discussed in terms of prediction of specific modes of subjective perception of illness and its practical implications for correction of patient's attitude to his/her disease, correction of non-compliance, optimization of therapeutical alliance and use of heart care resources. PMID- 10666883 TI - [Therapy of autonomic disorders by xanax (alprazolam)]. AB - The paper presents an open noncomparative investigation of 36 patients with different manifestations of the syndrome of autonomic dystonia. 20 patients (group 1) had permanent autonomic disorder in context of generalyzed anxious disorders, 16 patients (group 2) had panic attacks. The examination was performed before and 4 weeks after monotherapy with xanax (1.5-2.5 mg/day). Clinical neurologic study estimated both presence and a degree of manifestations of the syndrome of autonomic dysfunction, hyperventilatory syndrome and sleep disorders. Psychologic investigation included estimation of anxiety according to Spilberg's test, depression according to Beck's scale; SCL Scale was also used. Algesic syndrome was estimated by complex algesic questionnaire. Neurophysiologic study determined a contingent negative deviation and nociceptive flexory reflex. A positive therapeutic activity of xanax was established. The highest therapeutic effect was achieved in group 1 (83%) using lower doses (1.5 mg/day). In group 2 higher doses were needed (2.5 mg/day). In this case the effect was achieved in 83% of the cases, but full absence of panic attacks was observed only in 25% of the patients. Predictors of the drug's efficiency appeared to be short duration of the disease, slight manifestation of depression and absence of the algesic syndrome. PMID- 10666884 TI - [24-h ambulatory monitoring of cardiovascular indices in patients with panic disorders]. AB - 24-h monitoring of arterial pressure (AD) and heart rate (HR) was twice performed in 14 patients with panic attacks (in exacerbation and after a course of fluoxetine for 6 weeks). Panic attacks were followed by objective increase of both AD and HR in only 1/3 of the cases and by objective HR increase in 60% of the patients. Although patients with panic attacks were normothymics, hypertensive reactions can arise in them, during sleep for the most part. Alteration of HR variability in sleep-awakening cycle was the most paradoxical (excess HR variability in awakening state and decreased change during a sleep). These alterations of HR variability persisted after control of panic attacks with fluoxetine. This allows to consider HR variability as a factor which reflects predispositional peculiarities of autonomic system functioning in patients with panic disorders. PMID- 10666885 TI - [Neurophysiologic mechanisms of the disorders in recognition of emotions in endogenic depression]. AB - In 46 male patients with endogenic depressions of different nosology (22 patients with affective disorder, 24 with juvenile slow-progredient schizophrenia) and 22 healthy individuals (a control group) visual evoked potentials (EPs) were registered during recognition of emotionally positive, negative or neutral facial expression. It was found that deficient recognition of emotions in depressions was determined by alteration of different stages of information analysis. In inferio-temporal and occipetal cortical areas the Eps revealed delayed primary analisis of visual information and reduction of components responsible for classification stage and decision making. In anteriocentral cortical areas only late stages of analysis were altered, and they were depending on nosology of depressive sindrom. Dynamics of topographic maps revealed disturbance of analysis in the right hemisphere in patients with affective disorders and in left hemisphere in schisophrenics. In both groups of patients deficit of interhemispheric transfere was found. PMID- 10666886 TI - [Sporadic amyotrophic lateral sclerosis associated with Asp90Ala CuZn-superoxide dismutase mutations in Russia]. AB - 16 blood samples from Russian patients (Moscow) with idiopathic motor neuron disease were analysed for mutations in the CuZn-superoxide dismutase gene. Two patients (12.5%) with amyotrophic lateral sclerosis (ALS) were found to have a disease related mutation. One patient appears to have autosomal recessive adult onset ALS associated with homozygosity for Asp90Ala and presents the characteristic phenotype of very slowly ascending paresis with both lower and upper motor neuron signs. The other patient heterozygous for Asp90Ala presents ALS with lumbar onset and rapid progression. Both of cases are apparently sporadic. PMID- 10666887 TI - [The level of middle mass molecules and lipid peroxidation in blood of patients with different forms of stroke]. AB - To elucidate correlation between processes of lipid peroxidation (LP) and accumulation of peptides from the group of "middle molecules" (PMM) in blood, we made investigation of blood of 81 patients with ischemic and hemorrhagic stroke. Gel-filtration column chromatography for PMM separation was performed. Divided fractions were registered with UV-monitor at wave length equal to 254 nm. The character of both spontaneous and Fe(2+)-induced chemiluminescence, the level of malonic dialdehyde (MDA) and conjugated dienes (CD) were estimated. It was established that the level of LP processes was significantly increased in blood and erythrocytes of patients with stroke, especially in hemorrhagic form. The total content of PMM, especially of A and A4 fractions, as well as D and F fractions was almost twice as much in blood. A correlation was observed between accumulation of PMM in blood of patients with stroke and PL induction, that may be the factor of PMM generation. A conclusion was made, that accumulation of some PMM classes in blood of patients with strokes was related to LP processes, which were responsible for the severity of the secondary pathochemical alterations in tissues of the brain. These findings should be considered in elaboration of drug treatment and estimation of the disease prognosis. PMID- 10666888 TI - [Peculiarities of polymorphism in gene of serotonin transporter in men from different ethnic groups with acute alcoholic psychoses]. AB - Analysis of polymorphism in locus of the gene of serotonin transporter (hSERT) was performed in men of Russian and Tatar nationalities with acute alcoholic psychoses by polymerase chain reaction. An absence of the differences in the distribution of the frequences of genotypes and alleles of hSERT gene was observed between populations examined, that corresponded to such values in Caucasians. An association was found between the allelic variations with 9 units of the repetitions of hSERT gene and an early onset of chronic alcoholization and development of acute alcoholic psychosis. Among the men, who developed acute alcoholic psychosis at the age over 35 there were significant differences in distribution of frequences of genotype of hSERT-gene between the populations. Genetic heterogeneity of such pathology in Russians and Tatars and impact of hSERT in the development of the disease in Tatars, are suggested. PMID- 10666889 TI - [Factors of lonely living in old psychiatric patients]. AB - 70 lonely patients 60-83 years old were examined. Loneliness in the disintegrated first group (54 patients) resulted from the disintegrated family (the patients were early married). The loneliness in the second group was determined by the absence of their own family (16 patients). The main factors leading to loneliness were such diseases as schizophrenia and epilepsy, paranoic disorders of persecutive character directed to the nearest family, hypochondric depression, psychopathic-like disturbances, mental defect, personality disorders in premorbid state. Among social-psychologic factors the following ones had a significance: underestimation of the mental disorders in patients by their relatives, conflicts in the family, the patient's negative orientation to marriage, their deep emotional interrelation with the parents in young and mature age, alcoholism of married couple and special psychopathic features of the relatives. The events relating with the old age (loss of elderly couple), separation with adult children and parents, bad domestic living conditions were of less significance. PMID- 10666890 TI - [Classification of the dementias]. PMID- 10666891 TI - [Diagnosis of celiac disease]. PMID- 10666892 TI - [Rational management of acute community-acquired pneumonia]. PMID- 10666893 TI - [Transgenic food]. PMID- 10666894 TI - [The pediatrician and children from immigrant families]. PMID- 10666895 TI - [Recommendations for the dietary treatment of phenylketonuria]. PMID- 10666896 TI - [Body fat mass in boys and male adolescents]. AB - OBJECTIVE: To describe the proportion of body fat mass, quantified from the measurement of 4 skinfold thickness, in a sample of male children of Zaragoza. METHODS: In 1995, we have studied 701 males aged 6.0 to 14.9 years, from 6 schools of the province of Zaragoza. We have measured: weight, height, and biceps, triceps, subscapular and suprailiac skinfold thicknesses. Body density has been calculated from the 4 skinfold measurements, with the formulas of Sarria et al. Fat mass (%), was calculated with the Weststrate and Deurenberg's formulas. In each age group, we present mean, standard deviation and 5, 25, 50, 75 and 95th percentiles. RESULTS: We present mean, standard deviation and 5, 25, 50, 75 and 95th percentiles of body fat mass (%), in each age group. Body fat mass (%) progressively increased until 10.5 years and, after, progressively decrease at least until 14.5 years. CONCLUSIONS: We present data of body fat mass (%), calculated from the measurement of 4 skinfold thicknesses, obtained from a reference population in the province of Zaragoza. These data are useful as more rationale criteria in the diagnosis of obesity in children and adolescents. PMID- 10666897 TI - [Lupus anticoagulant in children. Report of 4 cases]. AB - OBJECTIVE: We present four cases of lupus anticoagulant (AL) in children. In addition, as a result of the lack of literature published on the subject in our country. We also evaluate the epidemiological, clinical and prognostic characteristics of AL in children. PATIENTS AND METHODS: The diagnostic criteria established by the "Subcommittee for the Standardization of Lupus Anticoagulant" were followed. RESULTS: Over a ten-year period (1988-1998), 46 cases of children with TTPA prolongation were documented. Nine children showed circulating anticoagulant, 4 of which were lupus type anticoagulants. The age of the patients ranged from 4 to 13 and there was a prevalence of males (3/4). Half of the children had a family history of bleeding dyscrasia and it was these who showed hemorrhage or thrombotic symptoms. The case that started with ecchymosis and hemorrhaging showed prothrombin prolongation due to factor II deficit. In 3 of the 4 children, AL was linked to acute respiratory infections and was transitory. The other coincided with two thrombotic episodes of the lower extremities in a healthy child. Positive anticardiolipin antibodies were detected in two patients, both showing repeated AL episodes, one with thrombosis and the other always asymptomatic. CONCLUSIONS: Diagnosis of AL in children is difficult and has probably been underestimated. Although it is usually transitory, it can appear in repeated episodes. Its early detection is important as it can be linked to both prothrombin deficit, as well as significant symptoms of hemorrhaging. PMID- 10666898 TI - [Nutritional status assessment in children entering foster care]. AB - OBJECTIVE: The purpose of this study was to assess the nutritional status of abused and neglected children in a middle-sized city (Zaragoza, Spain) at the time of entry into foster care. PATIENTS AND METHODS: A cross-sectional study was performed over a 7-year period. Using the anthropometric method, the nutritional status of 684 children (379 boys/305 girls), ranging in age from 1-17 years, was assessed. Weight, height and body-mass index were formulated (mean and DS) and compared with normal national reference standards (Z-score). Significant statistical differences were assessed (Student's unpaired t test). RESULTS: Below, arranged by age and sex, the significant variances reflecting subnormal standards are listed. Weight: 1 and 2 year old boys and girls (p < 0.001); 3 and 4 year old girls (p < 0.01); 3, 4, 5 and 6 year old boys and 7, 8 and 10 year old girls (p < 0.05). Height: 1, 5 and 9 year old boys and 1, 2, 3, 4 and 11 year old girls (p < 0.001); 2 and 4 year old boys and 6 and 10 year old girls (p < 0.01); 3, 6, 10, 15, 16 and 17 year old boys and 8, 9, 11, 12, 13 and 17 year old girls (p < 0.05). Body-mass index: 1 year old boys (p < 0.001); 6 year old boys (p < 0.01); 1, 2 and 7 year old girls (p < 0.05). CONCLUSIONS: Malnutrition and growth delay are present in a high rate of children entering foster care. In preschool children, moderate forms of acute and chronic malnutrition with wasting and stunting are predominant, identifying with the "failure to thrive syndrome". In primary and secondary school aged children, mild forms of chronic malnutrition with growth failure are predominant. When growth and developmental delay is present in children entering foster care, in the majority of cases, nutritional and emotional deficiencies are the etiology for this delay. This is consistent with the concept of "growth and developmental delay of psychosocial deprivation origin". PMID- 10666899 TI - [Calcium intake among school children in Badajoz]. AB - BACKGROUND: The intake of calcium has been studied in the diet of adolescents from a school in Badajoz (Spain) determining their BMI and their living habits (sport, consumption of tobacco and alcohol). METHODOLOGY: By means of an aleatory sampling, a sample n was selected = 207 (49.76% males and 50.24 females) with an average age of 14.14 +/- 2.08 years. They were weighed and measured to calculate their BMI. A questionnaire was carried out on consumption of products rich in calcium: milk and dairy produce, vegetables, fruit, chied fruits. The statistical analysis was carried out by means of Student t and ANOVA. RESULTS: The results was BMI < 25 (thin or standard) in 86.47%; BMI 25-30 (overweight) in 10.63% and BMI > 30 (overweight) 2.9%. The consumption of daily calcium was of 1304 +/- 702 g/ppd, higher (p < 0.001) in boys (10.45 +/- 5.51) than in girls (7.82 +/- 3.84). There is not correlation between BMI and calcium consumption. The boys observe the NIH recommendations. But girls usually don't. Milk products provide 87% of the consumed calcium. The boys who practice some sports consume 9.88 +/- 5.23 g/pps, the other ones 7.09 +/- 2.95 g/pps. CONCLUSIONS: 1. The weekly intake of calcium by surveyed students is 9.13 g/pps, smaller in girls than in boys. 2. Milk products provide 87% of the consumed calcium. 3. A 10.63% of them is overweight and a 2.9% is very overweight. 4. A 10.63% usually smokes and a 20.29% consumes alcohol. 5. A 26.27% doesn't practice any sports and consumes less calcium (p < 0.001) than the recommended quantity. PMID- 10666900 TI - [Forearm bone mineral density in healthy children]. AB - OBJECTIVE: In order to establish the normal patterns of forearm bone mineral density (BMD), BMD in the cortical and trabecular parts of the distal forearm were studied in a normal pediatric population. PATIENTS AND METHODS: BMD was measured by dual-energy X-ray absorptiometry (HOLOGIC QDR-1000) in the distal third forearm of 246 normal children and adolescents (111 boys and 135 girls) ranging from 2.8 to 20.8 years of age. BMD was correlated by multiple regression analysis with age, weight, body mass index (BMI), sex and pubertal Tanner stage. RESULTS: Forearm BMD increased progressively with age, weight, height, BMI and maturity, with the maximal increase in all forearm bone sites occurring at the onset of puberty in girls and boys. A statistically significant correlation was found between forearm BMD and all of these variables (r = 0.65 to 0.92). Mean BMD was higher in boys than in girls in cortical, trabecular and cortical-trabecular sites of the distal forearm. Maximal differences in BMD between boys and girls occurred at 17-18 years of age, especially the trabecular-dominated (ultradistal) part of the distal forearm (0.446 vs 0.384 g/cm2). CONCLUSIONS: Forearm BMD studies permit information of both cortical and trabecular bone mineralization to be obtained at the same time. This study reports normative data for forearm BMD in a healthy pediatric population. The values obtained may be used as a reference of normality when evaluating bone density in situations where skeletal mineralization may be compromised. PMID- 10666901 TI - [Infectious mononucleosis: study on hospitalized children]. AB - OBJECTIVES: To analyze the patients diagnosed of infectious mononucleosis (IM) in our institution, in order to study the symptoms and clinical evolution. MATERIAL AND METHOD: We retrospectively analyse 37 hospitalized children aged between 3 months and 14 years. Diagnosed of IM by production of immunoglobulin M (IgM) to viral capsid antigen (VCA). RESULT: In our series of 37 children, 14 were less than 4 years old. Among clinical data, lymphadenopathy (86%), fever (81%) and pharyngotonsillitis (70%) were the more striking. We found jaundice only in three of the older children. Upper airways obstruction was common, and specially severe in young children. Three patients developed pneumonia during the disease course. Only ten patients produced heterophile antibodies. Two children showed dual antibody rises to Epstein-Barr virus and cytomegalovirus. Corticotherapy was used in 7 children. The complications are presented in 68% of patients. CONCLUSIONS: Clinical aspects of IM were similar to those in the young adult, and complications occur more frequently. We found lack of heterophile antibodies, more frequent in youngest. PMID- 10666902 TI - [Apert syndrome: clinico-epidemiological analysis of a series of consecutive cases in Spain]. AB - OBJECTIVE: Apert syndrome is one of the five craniosynostosis syndromes caused by allelic mutations of the fibroblast growth-factor receptor 2 (FGFR2). It is characterized by symmetrical cutaneous and bony syndactyly of the hands and feet and a variety of pleiotrophic features of the skeleton, central nervous system, skin and internal organs. PATIENTS AND METHODS: We show the clinical and epidemiological characteristics of the 17 cases of Apert syndrome identified in a consecutive series of 26,956 malformed liveborn infants detected among 1,502,639 livebirths surveyed by the Spanish Collaborative Study of Congenital Malformations (CEMC) between April 1976 and March 1998. RESULTS AND CONCLUSIONS: The estimated frequency of Apert syndrome in Spain is 0.11 per 10,000 liveborn infants. All of the cases were sporadic and were associated with an increased paternal age. The clinical manifestations of our cases are concordant with the variable expression of the syndrome, with the cardinal features of acrocephaly secondary to craniosynostosis and syndactyly of hands and feet present in all cases, and other anomalies, including cardiovascular (23.5%), cleft palate (23.5%), urinary (5.9%) and central nervous system (5.9%), in some of the patients. PMID- 10666903 TI - [Extracorporeal membrane oxygenation, ECMO. Experience with the first 22 cases]. AB - OBJECTIVE: Extracorporal membrane oxygenation (ECMO) is an alternative to cases of respiratory or cardiopulmonary insufficiency when conventional therapy has failed. We present the first 22 patients treated with ECMO at the neonatology unit of the "Gregorio Maranon" Hospital. PATIENTS AND METHODS: From October 1997 until September 1999, 22 patients were treated with ECMO. In 8 of them ECMO was necessary because of respiratory insufficiency without response to conventional treatment (r-ECMO) and a veno-venous tidal flow system was used. In 14 patients, cardiac ECMO was necessary in the veno-arterial modality because of ventricular failure after extracorporal circulatory assistance during cardiovascular surgery. RESULTS: The 8 patients of the respiratory ECMO group had a mean previous oxygenation index of 89 +/- 36.6 (50-150). The mean duration of ECMO was 7.8 +/- 6.5 (1-16) days and the age at the beginning of ECMO ranged between 1 and 151 days. The most frequent indications in this group were congenital diaphragmatic hernia in three cases, meconium aspiration syndrome in 2 cases and 1 case each of septic shock, idiopathic pulmonary hypertension and air leak. The cannula was removed in 5/8 patients due to recovery and 5/8 survived. The 14 patients of the cardiac ECMO group needed veno-arterial ECMO because of severe ventricular failure. Eight out of fourteen were decannulated after improvement. In 5/14 there was multiorgan failure or bad neurological prognosis resulting in death while on ECMO. In 1 out of 14 patients removal of the cannula was impossible because of cardiac insufficiency. The mean age at the beginning of ECMO was 54 (3-178) days. The mean weight at ECMO was 3209 +/- 739 (2700-5000) gr and the mean duration of ECMO was 6 (1-15) days. CONCLUSIONS: ECMO is an effective treatment in rescuing critical patients when conventional treatment fails. Meconium aspiration syndrome is the pathology with the best prognosis on ECMO. Cardiac ECMO represents a complex group of patients in which ECMO is the only treatment and which may result in recovery in 40% of the patients. PMID- 10666904 TI - [Eosinophilic meningitis of unknown etiology. Infrequent occurrence in pediatrics]. PMID- 10666905 TI - [Type I neurofibromatosis associated with internal carotid artery hypoplasia]. PMID- 10666906 TI - [Generalized hypopituitarism in absence of hypophyseal stalk with ectopic neurohypophysis in a tall-for-age girl]. PMID- 10666907 TI - [Neonatal macrocephaly caused by multiple cerebral arteriovenous fistula treated with embolization techniques]. PMID- 10666908 TI - [Neurotuberculosis: multiple tuberculomas in a 6-year-old child]. PMID- 10666909 TI - [Anemia in a girl from the Antilles]. PMID- 10666910 TI - [Recommendations for basic, advanced, and neonatal cardiopulmonary resuscitation. IV. Resuscitation in newborns. Spanish Group for Pediatric and Neonatal CPR]. PMID- 10666911 TI - [Transient benign persistent hyperphosphatasemia in children]. PMID- 10666912 TI - [Value of the dissemination of the Mulford Library catalog in pediatrics]. PMID- 10666913 TI - [Transverse HIV myelitis in a child with AIDS]. PMID- 10666914 TI - [Cholestasis associated with parenteral nutrition in a critically ill child]. PMID- 10666915 TI - [Regulation of anaphylactic responses by Fc receptors]. PMID- 10666916 TI - [Nasal allergy--its immunological view]. PMID- 10666917 TI - [Skin management of itching in atopic dermatitis]. PMID- 10666918 TI - [Keeping dogs indoor aggravates infantile atopic dermatitis]. AB - We had a two-month-old girl with severe dermatitis since birth. Her serum RAST to HD, Df and Dp were 1.06, 0.03 and 0.01 Ua/ml respectively. A Yorkshire terrier were kept at her mother's parents' home where the patient had lived for a month since birth. Her eczema, which became markedly aggravated whenever she visited there, improved after the elimination of the dog. We investigated the relationship between keeping dogs and infantile atopic dermatitis. We studied 368 patients under the age of two years (211 boys and 157 girls). Skin symptoms were graded globally mild, moderate or severe. Total serum IgE and specific antibody titer to dog dander were measured. We asked them whether they kept dogs and specifically, where they kept dogs, outdoor, indoor, in their own house, or in their grandparents' house. 197 patients had no contact with dogs, 90 patients kept dogs outdoor and 81 patients did indoor. The positive rate of RAST (> or = 0.7 Ua/ml) to dog dander was 6.1%, 17.8% and 46.9% respectively in these three groups. There were strong statistical differences between three groups. On the other hand, among the 81 patients who kept indoor, the RAST positive rates were almost same regarding where the dogs were kept, in their own house or their grandparents' house. Interestingly this difference happens only with patients under the age of 3 months. Patients older than 4 months showed no significant differences in the positive RAST rates, whether they kept dogs indoor or outdoor. This suggests the sensitization occurs before the age of 3 months. Speaking of symptoms, patients who kept dogs indoor showed significantly more severe symptoms than patients who had no contact with dogs and patients who kept dogs outdoor. There was no significant difference between the symptoms of patients who had no contact with dogs and those of patients who kept dogs outdoor. This implies the patient's symptom will improve only by moving the dog out of the house. PMID- 10666919 TI - [Clinical significance of allergen specific immunotherapy in adult house-dust mite-sensitive bronchial asthma: impact on disease severity and medical cost]. AB - The objective of this study was to evaluate the clinical significance of allergen specific immunotherapy in house-dust-mite-sensitive adult bronchial asthma. Fifty patients treated with rush immunotherapy using house dust antigen were examined. The disease severity was compared between before and a year after the maintenance immunotherapy: reduction in the severity was observed in 27 patients (54.0%) following the treatment. The response rate was greater in the patients with step 3 (moderate, persistent) or step 4 (severe, persistent), disease period less than 10 years, or reversible airway obstruction. Patients who showed favorable clinical response also demonstrated the reduction in medical costs. These results suggest that allergen immunotherapy reduces the disease severity and medical cost in a certain population of adult atopic asthma. PMID- 10666920 TI - [A mechanism for the anti-inflammatory effect of nedocromil; inhibition of both adhesion molecule expression on eosinophils and endothelial cells, and eosinophil chemotactic activities]. AB - The accumulation of eosinophils in the airway is one of the characteristics seen in patients with bronchial asthma. One of the newly developed anti-asthma drugs (controller), nedocromil sodium (nedocromil) is known to suppress the influx of eosinophils into allergic lesions. However, little is known about this mechanism. Therefore, in this report we investigated the effects of nedocromil on Mac-1 expression on PAF-stimulated eosinophils, and adhesion molecule expression on endothelial cells stimulated by either IL-1 beta or IL-4. We also investigated the eosinophil chemotaxis. A significant suppression of the Mac-1 expression on PAF-induced eosinophils was observed at both concentrations of 10(-5) and 10(-7) M of nedocromil. The expression of adhesion molecules, particularly ICAM-1 and E selectin, on IL-1 beta-stimulated human umbilical vascular endothelial cells (HUVEC) was significantly suppressed at these concentrations, whereas the VCAM-1 expression was not changed. No significant suppression of VCAM-1 expression on IL 4-stimulated HUVEC was observed, although there was a tendency of suppression at these concentrations. On the other hand, the expression of the E-selectin molecule was significantly suppressed by nedocromil even under resting (non stimulated) condition. PAF-induced eosinophil chemotactic activities were also suppressed at these concentrations in a dose-dependent manner. These results suggested that nedocromil suppressed the influx of eosinophils to inflammatory lesions by inhibiting not only the expression of the Mac-1 on eosinophils and of E-selectin and ICAM-1 molecules on HUVEC, but also the eosinophil chemotactic activities. PMID- 10666921 TI - [Effect of rush immunotherapy (RIT) on Hymenoptera allergy]. AB - In our country approximately forty people die every year from anaphylaxis caused by hymenoptera stings. Between 1988 and 1996, 48 patients, who had experienced a systemic reaction to hymenoptera sting and were proved to have specific IgE antibodies to wasp, yellow or both (RAST score > or = 2), received rush immunotherapy (RIT) using venom extracts in our hospital. Fifteen patients had re sting after RIT. Fourteen out of the 15 patients showed only local reaction to the hymenoptera re-sting and one patient had mild generalized symptoms. Although one patient showed mild generalized uriticaria during RIT, no adverse reaction occurred during and after RIT in the other subjects. Follow-up studies on the titers of serum total IgE antibodies and hymenoptera specific IgE and IgG4 antibodies revealed that total and specific IgE antibodies transiently increased one month after RIT and returned to their baseline values by 6 months after RIT, while specific IgG4 antibodies continued to gradually increase up to al least 3 years after RIT. These results demonstrates that RIT is effective in prevention of a systemic reaction to hymenoptera re-sting and an increase in the titer of hymenoptera specific IgG4 antibodies may at least partly explain the efficacy of RIT. PMID- 10666922 TI - [Assumption of the area supplying Okayama Prefecture with Cryptomeria japonica and Cupressaceae airborne pollen, and of their scattering routes]. AB - It is very important to predict and disseminate information about the total pollen counts of both Cryptomeria japonica and Cupressaceae for patients with pollinosis. In Okayama Prefecture, we have reported that the pollen counts of both Cryptomeria japonica and Cupressaceae are influenced by the meteorological conditions in the previous July. We predicted the area supplying Okayama Prefecture with Cryptomeria japonica and Cupressaceae pollen, and also the route of airborne pollen from the meteorological conditions and a topographical map of Okayama and four neighboring prefectures. It was found that Cryptomeria japonica and Cupressaceae pollen counts at the four observation sites correlated very well with the meteorological conditions at Tsuyama weather station in Okayama prefecture. Therefore, we considered that the areas supplying Okayama prefecture with Cryptomeria japonica and Cupressaceae pollen were the central northern areas including Tsuyama, 85.7% of whose plantation areas contained Cryptomeria japonica and Chamaecyparis obtusa, and that their pollen was carried along the routes of three major rivers, R. Takahashi, R. Asahi and R. Yoshii. PMID- 10666923 TI - [Effectiveness of preventive activities]. PMID- 10666924 TI - [PAPPS effectiveness study (1998) and preliminary results of the PAPPS evaluation (1999)]. PMID- 10666925 TI - [Prevention and health promotion in childhood and adolescence. Group for Prevention in Childhood and Adolescence of the PAPPS]. PMID- 10666926 TI - [Preventive cardiovascular recommendations: practical applications of cardiovascular risk. Group for Cardiovascular Prevention of the PAPPS]. PMID- 10666927 TI - [Cancer prevention. Group for Cancer Prevention of the PAPPS. Update 1999]. PMID- 10666928 TI - [Prevention of transmissible diseases. Work Group for Transmissible Diseases of the PAPPS]. PMID- 10666929 TI - [Recommendations on life style. Work Group for Health Education and Cardiopulmonary Prevention]. PMID- 10666930 TI - [Prevention of mental health disorders in primary health care. Group for Prevention in Mental Health of the PAPPS]. PMID- 10666931 TI - [Cutaneous asthenia (Ehlers-Danlos syndrome) in a domestic rabbit]. AB - Cutaneous asthenia is a connective tissue disease primarily of dogs and cats, resembling Ehlers-Danlos syndrome in man. This is a description of the disorder in a rabbit. The one-year-old female animal was presented because of two large gaping wounds of the skin. Clinical examination revealed a hyperextensible, thin, and fragile skin. The degree of skin extensibility was evaluated by means of a skin extensibility index (SEI: 19.2%) and compared with those of 4 healthy rabbits (SEI: 8.3%-14.3%). Clinical diagnosis was confirmed by histopathological examination of a skin biopsy revealing reduced packing density of collagen fibers. In addition a decreased number of hair follicles was observed. No conclusion could be drawn regarding the etiology, but existing literature strongly suggests a genetic cause (mostly autosomal dominant inheritance) in humans as well as in animals. PMID- 10666932 TI - [Fluctuations of urea, cholesterol and triglyceride concentrations in plasma of clinically health piglets during the first four weeks of life]. AB - The present study was designed to investigate fluctuations of plasma concentrations in urea, cholesterol and triglyceride in healthy piglets (Landrace x Pietrain) during the first four weeks of life. Blood samples were taken by venipuncture in 240 piglets thirty minutes after birth, after 6, 24, 48, 168, 336, 504 and 672 hours. Cholesterol and triglyceride concentrations exhibited minimum values after delivery and rose up to the 48th hour of life, what can be explained by colostrum intake. Although an increase of urea after birth was determined, but some of the following measured concentrations were under the initial value. The three parameters showed a significant dependency on the age (p < 0.001). After 6 hours there was a correlation of cholesterol concentration and sex (p = 0.001) and triglyceride concentrations have been correlated to body temperature after 24 and 48 hours (p = 0.009; p = 0.004). Before colostrum intake urea concentration has been associated to body temperature (0.001). The physiological significance of the observed fluctuations for adaptation from an intrauterine to an extrauterine life is discussed. PMID- 10666933 TI - [Quality of development and adaptation reactions of dairy calves at specific age periods in early life. Effect of rearing variations on proteins and minerals and on metabolic variables of blood]. AB - Calves from the dairy herd of the institute reared in groups with changing (Gw, N 16, m 9, w 7) and constant composition (Gk, N 16, m 9, w 7) since the first day of life and single box reared calves (E, N 11, m 7, w 4) were tested at 15, 30, 60 and 90 days of age. Venous blood samples were analysed for total protein, albumin, creatinine, blood urea, glucose, Ca, Mg, P and iron. At 15 days between group differences of mean values existed for total protein, albumin, blood urea, creatinine and at 30 days for rectal temperature, albumin, blood urea, Ca, P and Fe, at 60 days for blood urea, glucose, Ca, Mg, P and at 90 days for total protein, albumin, creatinine, glucose, Mg and P. Interactions between group and gender could not be demonstrated. Significant changes of variables with age were most frequent in calves of group Gk, and more frequent with creatinine, blood urea, Fe and glucose. Creatinine concentration diminished permanently with age of the animals, that one of total protein and albumin increased moderately. Blood urea was higher at 90 days, and serum iron at 60 and 90 days than before. Blood glucose concentration at 90 days was lower in calves of group Gw and Gk comparing group E. Significant group differences appeared at 15 days (total protein, albumin, blood urea, creatinine) and remained till 60 days (blood urea) and 90 days (total protein, albumin) or became stronger at 90 days (creatinine). Total protein, albumin and blood urea were higher in group E than in group Gk and Gw calves. Group Gw calves had smallest glucose concentrations at 60 and 90 days. Significant between groups differences for Ca, Mg, P existed for all sampling points, those for serum iron at 30 days only. Greater Ca and Mg values were found in group Gk and group E calves, greater P concentrations up to 60 days of age in group E calves. P concentrations changed differently in group Gk and group Gw calves with age. Creatinine showed moderately high between sampling point correlations. Changes of variables in calves between age points in relation to the starting situation at 15 days of age showed significant correlations in many cases meaning that there were directed individual and time specific adaptation processes. PMID- 10666934 TI - Efficacy of a combination of amoxicillin and clavulanic acid in the treatment of pneumonia of pigs. AB - Amoksiklav was used in the therapy of mixed respiratory tract infections in weaned pigs under field conditions. Positive effects of therapy with Amoxicillin and Clavulanic acid were observed in the majority of treated pigs. The production losses due to pneumonia in pigs treated with this combination were lower than among control pigs treated with Oxytetracycline, also a significantly lower ratio of death was observed among experimental weaners in comparison to the controls. Thirty days after the end of the therapy it was found that the body weight gain (b. w. g) of the experimental animals was on average 800 g higher after this period and the experimental piglets grew daily in average 20 g more than the controls. Experimental pigs were slaughtered 3 days earlier than the controls and the average weight gain at slaughter was highest by 1.1 kg in this group. Average daily b. w. g. of experimental pigs during the period from birth to slaughter was 13 g higher in comparison to the controls. PMID- 10666935 TI - [Orienting endotoxin measurement in the atmosphere]. AB - Airborne endotoxins are supposed to influence the respiratory health of animal and man in animal housings as well as at certain work places in agriculture. Little is known about the usual concentrations of endotoxin in the outdoor atmosphere. Therefore in a field study 30 air samples were taken, 22 samples in a rural region, and 8 in a more residential and industrial area. The samples were taken by impingement and filtration. The analysis was carried out by means of the chromogen-kinetic limulus amoebocyte lysate test (LAL-Test). The median concentration of all samples was 0.36 ng/m3. The highest median concentration of 0.49 ng/m3 was found in summer with a maximum value of 1.80 ng/m3 indicating large variations. In the other seasons the median concentrations (ng/m3) were distinctly lower (spring: 0.30, n = 11; autumn: 0.26, n = 5; winter: 0.19, n = 3). No significant differences were observed between farming and residential areas at this low concentration level. The concentrations of endotoxin found in this study were far below all threshold limits which are presently discussed for work places. PMID- 10666936 TI - Simultaneous transmission of Trypanosoma mukasai, Babesiosoma mariae and Cyrilia nili to fish by the leech Batracobdelloides tricarinata. AB - Trypanosoma mukasai (HOARE, 1932), Babesiosoma mariae (HOARE, 1930) and Cyrilia (= Haemogregarina) nili (WENYON, 1909) were concurrently transmitted from Tilapia nilotica to Clarias lazera using the leech vector Batracobdelloides tricarinata. Concurrent transmission was more successful in immature Clarias lazera in which prepatent periods were shorter and patency longer than in mature fish. PMID- 10666937 TI - [Possibilities and limits of assisted reproduction]. AB - The term assisted reproductive technologies (ART) describes all treatment procedures which include more than the natural intercourse to conceive. The most invasive procedure is intracytoplasmic sperm injection (ICSI), which was introduced in 1993. By ICSI the successful treatment of couples with severe male factor infertility, even in cases of azoospermia, became possible. In cases where only immature forms of spermatogenesis are present in the testicular tissue, pregnancies could also be achieved. The limiting factor with greatest influence on the success rate of ART procedures is the female's age. In this context the treatment of postmenopausal women by oocyte donation, as well as ooplasma donation to overcome the negative influence of the female's age on oocyte quality are extreme examples of ethically questionable ART procedures. Until now several 100,000 children worldwide have been born subsequent to ART and especially in vitro fertilisation. No increase in the malformation rate neither after IVF nor after IVF/ICSI have been reported so far. However, because of the lack of prospective controlled studies using standardised evaluation procedures and having enough statistical power to prove the security of ICSI for the newborn children, the German statutory health insurance bodies have ceased to pay ICSI costs as of July 1999. The data on the success of ICSI, the known data on the newborn and developing children and the decisions of the medical services of the German statutory health insurance bodies are presented in discussed in this paper. PMID- 10666938 TI - [Mother-child treatment. Chance for differentiated and specific introduction of necessary psychological preventative and treatment measures]. AB - Many women are often caught up in multi-layered situations of stress and strain without having an opportunity to speak about themselves and their difficulties. Within the setting of a health recovery programme for mothers and their child(ren), a first step might be possible to raise their awareness for an underlying psychological problem. Interventions might focus on symptoms pertaining to the psychological, psychosomatic and psychosocial sphere. Experience shows that the clients mostly present with problems specific to women. Describing various interventions, the author discusses psychological support measures that might be provided for women in the course of a health recovery programme. PMID- 10666939 TI - [Drug prescribing in primary health care for diabetic and non-diabetic patients: effect of therapeutic drug budgeting]. AB - With computerised data on drug prescriptions, which were collected among a sample of 362 internist, general and medical practitioners throughout Germany, the effects of the drug budget, based on the German Health Care Structure Reform Act (GSG), on diabetic patients were analysed. The data of 3053 diabetic patients (10% random sample) were compared with the same number of nondiabetic patients for the period of July 1992 to December 1994. The frequency of consultations per patient increased in both groups during the study period (p < 0.01). Diabetic patients had more contacts with the physicians than nondiabetic subjects (p < 0.01). The prescriptions and costs among the patients with diabetes decreased in the first six month of 1993 (-10%/ -16%). In the following time costs increased and exceeded the values of 1992 by about 13%. The costs per prescription decreased during the study period about 10%. The data show that costs as well as prescriptions per consultation between diabetic and nondiabetic patients remained in the same ratio. The proportion of consultations with > or = 1 prescription increased in diabetic patients after the GSG. A refusal to prescribe drugs in primary health care practices among diabetic patients was not observed. There was also no restriction on prescription drug use among diabetic patients compared with nondiabetics. PMID- 10666940 TI - [Ornithosis--studies in correlation with an outbreak]. AB - An outbreak of ornithosis with 8 cases of ornithosis pneumonia and 2 lethal complications was investigated in workers in a poultry farm and processing plant and a comparative seroepidemiological study of antigen responses was performed in 3 collectives: No. I: n = 82/87 workers in the processing plant, where the outbreak occurred; No. II: n = 83 workers in a chicken slaughter-house; No. III: n = 82 as matched-pair group to collective No. I with the same age and sex, but without occupational risk. The test systems were: genus specific complement fixation reaction (CFR), Ipazyme commercial slide kit containing LGV antigen and a type-specific microimmunofluorescence (MIF) technique with antigens binding C. psittaci, pneumonia and trachomatis IgA, IgG and IgM. 57/82 (71.9%) workers in group No. I were chlamydial antibody-positive, whereas only 22/82 of the population Nr. III--control group (odds ratio 6.2/3.2-12.3 p < or = 0.05). 16/83 (19.3%) of the workers in the chicken slaughterhouse had antibodies against chlamydia group antigens. 30/82 of the collective No. I had serological evidence of a recent or current infection with higher antibody titres in CFR and IPAZYME Test and/or antibody response against IgA and IgM (MIF). 43.3% of the latter could be serologically detected as specific infections with C. psittaci. 10 of 18 (55%) workers employed in the recent 3 months had serological signs of an acute infection. There was no association between the point of contact with the poultry (live hang areas, slaughtery, evisceration, cooling carcasses) and the prevalence of antibody response. The possible routes of infection, inhalation of dried excretions or aerosols and via hand-to-mouth contacts are discussed. In specimens of cloacal swabs and faeces of the ducks chlamydiae could be found although the animals were asymptomatic. The results of this study demonstrate that in poultry plants, where ducks and other poultry living in an aqueous habitat are slaughtered and processed, a high risk of C. psittaci infection (70.2%) and ornithosis morbidity (25%) with a lethality of 8.3% can exist. Since the eradication of C. psittaci in poultry does not seem to be possible at the moment, preventive measures e.g. gloves, masks, information and medical examinations of the workers must be implemented in those slaughterhouses and plants where C. psittaci is suspected or common. PMID- 10666941 TI - [Utilization of dental services--results of an oral hygiene study in Saxony]. AB - The utilisation of dental services is a precondition for professional preventive measures. The representative oral survey "Bevolkerungsreprasentative Studie zum zahnarztlich-prothetischen Versorgungsgrad und Behandlungsbedarf" aimed at gaining data on the frequency and motives for dental visits and the existence of barriers. The sample consisted of 714 subjects aged fifteen years and older. It was representative for the German Federal State Saxony. Socio-economic and behavioural data were collected via self-completed questionnaire. The caries history was evaluated by assessing the DMFT Index. Data collection in 1996 was carried out by three calibrated dentists. Utilisation of dental services was found to be high. Subjective barriers played no crucial role in the decision to utilise dental services. Costs as an objective barrier to dental visits had a growing importance with age. Lower rates of tooth loss and enhanced levels of restoration were found in subjects aged over 24 years who regularly utilised dental services compared to subjects with non-regular utilisation. These differences were significant for subjects over 54 years (tooth loss) and subjects aged 25-44 years and over 54 years (level of restoration). Compared to previous studies the utilisation was similar or higher in our sample. The enhanced oral health belief paves the way for professional preventive measures. In the over 64 year-olds the need for regular dental visits was obvious and this age group must definitely be given more relevant information on the subject. PMID- 10666942 TI - [Statistically evaluated 95% prediction ranges as alternative to reference ranges exemplified by polychlorinated dibenzo-p-dioxins/dibenzofurans and lead]. AB - Using the 95% prediction limits of an age-related multiplicative regression model describing the datasets of blood examinations carried out on subjects suspected of having been exposed to lead and to polychlorinated dibenzo-p dioxins/dibenzofurans (PCDD/F), it was shown that the relevant curves can be used as an alternative to reference ranges describing the actual background exposure to these pollutants. The upper limit of the actual German background exposure can be estimated by the following equations: PCDD/F as International Toxicity Equivalents in the age range of 10-70 years [pg/g lipid basis] = 1.64.age0.871 and lead in the age range of 15-80 years [microgram/l] = 18.15.age0.3638. PMID- 10666943 TI - [Public health today]. PMID- 10666944 TI - [Quality assurance and teamwork in social medicine expert assessment exemplified by the expert assessment aid "corrective breast surgery" of the North Rhine Health Insurance Medical Service]. AB - To standardise sociomedical expertising procedures in surgery of the breast a project team of medical advisors of the Medical Advisory and Expertising Service North Rhine worked out a manual. A form was created for sociomedical pre-advice occasions. Detail parameters for examination (time needed and advisor selection) were fixed in consent. Checklists were worked out, which may be admitted to quality assurance procedures. The manual "Medical opinion for surgery of the breast" has been implemented and is applied in the Medical Advisory and Expertising Service North Rhine since January 1999. In the sense of teamwork participation those who were involved as team-workers became participants in decision-making. The manual "Medical opinion for surgery of the breast" may be seen as an example of quality assurance and participation in socio-medical opinion procedures. The influence of this manual on the formal quality of medical opinion will be checked. PMID- 10666945 TI - [Examples of methodological problems in evaluating misuse of hospital admission and approaches to correction]. PMID- 10666946 TI - The role of telemedicine in medical education. PMID- 10666947 TI - Awareness of, use and perception of efficacy of alternative therapies by patients with inflammatory arthropathies. AB - Fifty one patients with chronic inflammatory polyarthritis were surveyed on unconventional treatments they used to self-treat their condition. Awareness of the availability of alternative therapies (ATs) was universal. Sixty-six percent (66%) of patients had tried one or more ATs. The most popular ATs were dietary manipulation (no red meat, dosing with vinegar and honey), the wearing of magnets and copper bracelets, and acupuncture. The best predictors of AT use were male sex, Caucasian race and formal education beyond high school. Numbers were too small to make definitive statements about perceptions of efficacy, but the users of magnets and fish oils tended to be dissatisfied with these ATs, while those who had tried bee stings, herbs and hormones claimed effectiveness. PMID- 10666948 TI - Trends across two time periods in the diagnosis of substance abuse comorbidity at the Hawaii State Hospital. AB - This study investigates the changes from the late 1980s to early 1990s of comorbidity (mental illness plus substance abuse) at the Hawaii State Hospital. For the 1990s, a prevalence rate ranging from 14.2% to 30% was estimated, with the latter figure based on a closer review of the records. A higher proportion of comorbid clients were single, and compared to the non-abusers (i.e., patients diagnosed with only schizophrenia or affective disorder), a higher percentage were male and had an educational level less than high school. There was an increase in the percent of non-abusers and substance abusers, but a decrease in the dual diagnosed. The implications of these findings are discussed. PMID- 10666949 TI - Clinical applications of hypnotherapy in a medical setting. AB - Since 1958, hypnosis has been recognized by the American Medical Association as a legitimate form of medical treatment when administered by an appropriately trained practitioner. With the prevalence of certification societies and international organizations, the specialty has increased its level of professionalism and clinical applications. However, in spite of increased exposure and utilization of this unique clinical application, its use within medical settings varies considerably. The purpose of this article is to provide an understanding of clinical hypnosis and offer clinical applications, with the goal of increasing its exposure and utilization within medical settings. PMID- 10666950 TI - [Morphometric analysis of nuclear variations in normal and neoplastic mammary ductal cells]. AB - Morphometry is a method to detect changes in a variety of tissues through quantitative elements. The purpose of this study was to examine several nuclear morphologic characteristics in normal and neoplastic mammary ductal cells using a multivariable method and expression of estrogen receptors by immunohistochemical techniques. A total of 1879 nuclei were examined by a computerized program, following the detection of estrogen receptors. Nuclear area, perimeter, diameter, maximal and minimal radio were obtained in 439 normal ductal nuclei. The mean nuclear area was 14.45 with a range between 10.88 and 17.90. Variables showed adequate statistical correlation (r > 0.5). A total of 1440 neoplastic nuclei were classified as grades I, II and III, and a statistical significative difference was found between these three groups. We conclude that the nuclear area is a reliable variable for statistical correlation being the ductal nuclei anisotropic objects. PMID- 10666951 TI - [Candida in biological human samples]. AB - Infections by Candida have been raising in the last decades, and risk factors, mainly immunosuppression and the appearance of Candida no albicans, are determinants in the prognosis of these mycoses. The purpose of this investigation was to identify and establish the prevalence of C. albicans and Candida spp. in candidiases, in patients to the Hospital Universitario de Maracaibo, whose biological samples were processed for both direct examination and cultures, needed for the proper identification. From October 1996 to October 1998, isolation and identification of yeasts of Candida were performed in 177 biological samples: 73 (41.24%) Candida albicans and 104 (58.75%) Candida spp. Both blastoconidias and pseudohyphae were found in 34 samples (19.21%), 24 of which (70.5%) were diagnosed as C. albicans and 10 (29.5%), as Candida spp. Blastoconidias identified by direct method were distributed as C. albicans 34.2% and Candida spp. 65.7%. C. albicans was found more often in intertrigo, sputum and in bronquial lavage samples. Candida spp. was more frequent in nails. Candidiasis is a frequently diagnosed mycosis in hospitals, mainly among immunossuppresed patients. It is very important to use direct microscopical evaluation and cultures, in order to establish the presence of blastoconidias and pseudohyphae, that will help to diagnose the aethiology and prevalence of candidiasis. It is also important to recognize subungueal candidiasis in hospital staff, that could spread the infection to inpatients. PMID- 10666952 TI - [Analysis of prevalence of point mutations in codon 12 of oncogene K-ras from non cancerous samples of cervical cytology positive for type 16 or 18 PVH]. AB - Ninety-one non cancerous samples from genital specimens positives for VPH 16 or 18 and 27 non-infected samples as controls were studied. Mutations at codon 12 in K-ras gene was analyzed using enriched alelic PCR technique. Among the samples studied 17.58% showed mutations in this codon. Significant differences were observed between the control group (negative DNA-HPV) and positives DNA-HPV samples (p < 0.01). No differences were found between both viral types in relation to the mutation frequency. The presence of mutations in the K-ras gene in non cancerous cytological samples point out new questions about the role of mutations in proto-oncogenes and the development of cervical cancer. PMID- 10666953 TI - Hypertrophic pulmonary osteoarthropathy in acquired immunodeficiency syndrome. Case report and review. AB - We describe a case of hypertrophic pulmonary osteoarthropathy (HPOA) in an adult patient with acquired immunodeficiency syndrome (AIDS). This is the ninth case of HPOA associated with AIDS in adults, reported in the literature. The presence of pulmonary tuberculosis was also suspected, based on clinical grounds. Cases of clubbing associated with AIDS infection are reviewed. PMID- 10666954 TI - [The NADPH-oxidase complex in chronic granulomatous disease: preliminary description of a cluster in Merida-Venezuela]. AB - Chronic Granulomatous Disease (CGD) is a primary immunodeficiency characterized by an unusual predisposition to develop bacterial and fungal infections due to a failure of phagocytic leukocytes to generate superoxide, required for the intracellular killing of microorganisms. The lack of superoxide production is secondary to a defect in the NADPH-oxidase enzymatic complex activation, as a result of mutations of any of the components. Both, X-linked and autosomal recessive patterns of inheritance have been demonstrated in this disease, being the X-linked the most frequent and characterized by mutations in gp91phox. Mutations in p47phox, p67phox and p22phox have been shown in the autosomal recessive pattern. The molecular and genetic characteristic of NADPH-oxidase complex and its pathology in CGD are reviewed along with a brief description of the preliminary findings in two families from Merida, Venezuela. PMID- 10666955 TI - Research assessment--a blessing in disguise? PMID- 10666956 TI - Intra-oral topical anaesthetics: a review. AB - OBJECTIVE: This review considers the use of topical anaesthetics in the mouth to reduce the discomfort of local anaesthetic injections and intra-oral operative procedures. DATA SOURCES: Electronic literature search using Pub Med, manual search of references within papers found by the primary search and manual searching of abstracts from scientific meetings. STUDY SELECTION: The articles selected for this review investigate or used topical anaesthesia in dental procedures. CONCLUSIONS: The use of topical anaesthetics does not guarantee pain free dental local anaesthesia. Efficacy is dependent upon the duration of application and the gauge of needle used. Evidence is available that the use of topical anaesthesia alone is sufficient to perform some intra-oral procedures including periodontal manipulations, operative dentistry and oral surgery. PMID- 10666957 TI - The teaching of Class I and Class II direct composite restorations in European dental schools. AB - OBJECTIVES: The purpose of this descriptive study was to provide updated data on the teaching of Class I and Class II direct composite restorations in Europe as part of a survey of this aspect of the primary dental curriculum in Europe and North America. METHODS: Data on the teaching of posterior composite restorations and related matters were collected by means of a postal questionnaire sent to 185 dental schools known to exist in Europe. Non-respondents were sent a second questionnaire after two months. Further information pertaining to student requirements was sought after six months from all respondents. RESULTS: The response ranged from 92% for dental schools in Scandinavia to 40% from dental schools in Southern Europe with an overall response of 56%. All but four of the 104 participating schools were found to teach the use of composites in Class I and Class II, two-surface situations in at least premolar teeth. Contraindications and techniques taught for posterior composites varied within and between the country groupings of Northern and Central Europe. Scandinavia, Southern Europe and Eastern Europe. However, certain consensus views were identified. The experience of adverse biological reactions to the use of resin based restoratives in European dental schools was found to be limited. CONCLUSIONS: Notwithstanding the variation in the response from the four geographic regions investigated and the relatively low overall response to the questionnaire, it is concluded that the data reported indicates that most dental schools in Europe teach the use of composites in selected Class I and Class II situations. However, considerable variation exists both within and between the regions investigated in relation to this teaching. Further research and consensus conferences should be planned to reduce variability across Europe in relation to the contraindications and techniques taught for posterior composites. PMID- 10666958 TI - Validity of the examination method of occlusal contact pattern relating to mandibular position. AB - OBJECTIVE: This study was based on the hypothesis that conflicting findings and inconclusive consensus regarding the role of occlusal factors in the masticatory system are due to the variations in the definitions and methods used to describe and examine the occlusal factors. The object of this study was to determine whether contact patterns during lateral movement vary with mandibular positions and whether the contact pattern in lateral positions close to the maximum intercuspation has characteristics distinct from those in an edge-to-edge position. METHODS: Occlusal contacts of 86 young adults were examined using shim stock in regulated lateral positions: 0.5, 1, 2 and 3 mm from the maximum intercuspation, where the 0.5, 1 and 2 mm positions were defined as lateral positions close to the maximum intercuspation and the 3 mm position as an edge-to edge position. RESULTS: The occlusal contact pattern in the 0.5 mm position showed a marked prevalence of posterior tooth contacts on the working and the non working sides, compared with the 1, 2 and 3 mm positions. The occlusal contact pattern in the 3 mm position predicted the presence or absence of the occlusal contact in the 1 and 2 mm positions (sensitivity > 0.7) but not in the 0.5 mm position (sensitivity < 0.6). CONCLUSION: The occlusal contact patterns during lateral movement varied greatly with mandibular positions. The examination method of the occlusal contact pattern in one unregulated position will be invalid. It is necessary to distinguish the occlusal contact patterns between a position close to the maximum intercuspation and an edge-to-edge position when investigating its role in the masticatory system and in oral disease. PMID- 10666959 TI - Investigations into the use of an ultrasonic chisel to cut bone. Part 1: Forces applied by clinicians. AB - OBJECTIVES: To measure in vitro the direction and force of applied loads applied by clinicians when using both a conventional slow surgical handpiece (CH) and an ultrasonic chisel (USC) for cutting bone. STUDY DESIGN: Five clinicians were asked to cut bovine bone using either an USC or a CH. The bone was placed on a force measurement system that could measure both longitudinal and downward loads. The rate of cut was calculated over a fixed time-period and the depth of cut measured using a penetratometer. RESULTS: The magnitude of the longitudinal forces generated varied between 1.48 and 3.22 N (USC) and 0.04 and 4.56 N (CH). The CH had a pulling force directed towards the operator. Both instruments produced a similar range of downward forces although there was intra- and inter operator variability. The rate of cut varied in a similar manner, however, the CH produced a significantly greater depth of cut (p < 0.05). CONCLUSIONS: The force measurement system demonstrated differences in the way clinicians used the USC and CH instruments to cut bone. Of the two cutting methods investigated, the rotary bur is more efficient than the ultrasonic chisel. An ultrasonic chisel does cut bone in a different manner from a conventional bur and clinicians may require training before using it clinically. PMID- 10666960 TI - Investigations into the use of an ultrasonic chisel to cut bone. Part 2: Cutting ability. AB - OBJECTIVES: Ultrasound may offer a possible alternative to rotary instruments for removing bone. This study was undertaken to analyse in vitro the various factors that influence the cutting of bone by an ultrasonic chisel. STUDY DESIGN: A block of bovine femur was moved in a longitudinal direction under a stationary ultrasonic chisel. The force and depth of the cut was recorded for cutting rates of 28-112 mm/min and with increasing rake angles of 0 to +20 degrees. The pressure exerted by the chisel was recorded for different cutting rates. RESULTS: When the cutting rate increases there is a corresponding increase in the downward force which is followed by a decrease in the force at rates greater than 56 mm/min. The depth of the cut increases up to a rate of 56 mm/min after which it decreases. Both the longitudinal and downward forces do not change when the rake angle changes from 0 to +10 degrees. The downward force decreases when the rake angle increases from +10 to +20 degrees. CONCLUSIONS: The bone is cut slowly with the ultrasonic chisel, but this would assist in precision. Where such an instrument is used for cutting bone the clinicians should be aware that both low forces and cutting rates are required, and the instrument should be held at a low rake angle. PMID- 10666961 TI - Composition of the oral streptococcal flora in healthy children. AB - OBJECTIVES: To identify the predominant streptococcal species in the mouths of healthy children and to investigate the composition of the oral streptococcal flora over a period of 4 months. PATIENTS AND METHODS: The subjects were 33 fit, healthy schoolchildren aged between 5 and 16 years. These children were part of a large study and were the matched controls for a group of subjects undergoing bone marrow transplantation. The oral flora was sampled using an oral rinse technique on two separate occasions 4 months apart. The outcome measures were the number of each streptococcal species per millilitre of oral rinse; the isolation frequency of each species; the proportion of each species as a percentage of both the total streptococcal count and the total anaerobic count. RESULTS: The predominant species were Streptococcus salivarius, S. oralis and S. mitis. There was no significant variation in the composition of the oral streptococcal flora over the 4 month period. CONCLUSIONS: The oral rinse technique provides a reliable method of sampling the streptococcal flora of children. PMID- 10666962 TI - Adaptation of dental plaque to metabolise maltitol compared with other sweeteners. AB - There is some evidence that plaque can adapt to regular exposure to some bulk sweeteners, leading to increased metabolism and acidogenic potential of the sweetener. This potential for adaptation varies between non-sugar sweeteners and has important implications for manufacturers of food, confectionery and medicines used long-term. Maltitol (99% purity crystalline D-maltitol) is a relatively newly approved non-sugar sweetener and appears to have potentially good dental properties. OBJECTIVES: To compare plaque adaptation to pure sucrose, sorbitol, xylitol or maltitol and the effect of their prolonged use on acid production by plaque from sucrose, in vivo. METHODS: Two series of plaque pH experiments were carried out. Each experiment involved a 14 day adaptive period when four 5 g lozenges of the sweetener were taken between meals each day. Each experiment was separated by a 14 day wash-out period. Acid production was quantified as: (a) minimum pH; and (b) cH area (difference between plaque pH curve and resting value, expressed as cH units). RESULTS: Thirteen adults, of mean age 41 years completed the study. When adaptation of dental plaque to the metabolism of sweeteners was compared, there was a statistically significant difference (p = 0.033) between xylitol and sorbitol, and between xylitol and sucrose but not between xylitol and maltitol. When the effect of prolonged use of sweeteners on acid production after sucrose rinsing was compared, there were no statistically significant differences between the sweeteners. CONCLUSION: Dental plaque does not adapt to metabolise xylitol or maltitol following prolonged exposure over 14 days. PMID- 10666963 TI - Cytotoxic effects of composite restorations employing self-etching primers or experimental antibacterial primers. AB - OBJECTIVE: The aim of this study was to investigate the cytotoxic effects of composites which employ proprietary self-etching primers or experimental primers containing an antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) on human pulpal cells by in vitro dentine barrier tests. METHODS: Experimental primers were prepared by the addition of MDPB to each of two control proprietary primers at 1, 2 and 5%. Direct and indirect composite specimens were placed using each primer on one side of a dentine disk assembled in a simple pulp chamber device. Human pulp cells were incubated on the other side of the disk. After 48 h of incubation, the uptake of [3H]-thymidine by the cells was compared with that for negative controls using wax. Tests were repeated four times for each material. The diffusion of monomers from each specimen was determined using the same device. RESULTS: The specimens with control primers showed 26-35% reduction in cell activity. There were no significant differences in cytotoxicity between the control and experimental primers-specimens (Kruskal-Wallis test, p > 0.05). 2-Hydroxyethyl methacrylate at more than 1 mg ml-1 diffused from all specimens and was considered to be the cause of cytotoxic effects. The concentrations of MDPB diffused from the experimental primers-specimens were less than the toxic level, even for 5% MDPB-containing specimens. CONCLUSIONS: The results demonstrate that composites employing proprietary self-etching primers produced cytotoxic effects on human pulpal cells in vitro, although the toxicity was not severe. Incorporation of MDPB into a proprietary primer of up to 5% had no significant influence on the cytotoxicity observed. PMID- 10666964 TI - Retentive and compressive strengths of modified zinc oxide-eugenol cements. AB - OBJECTIVES: This investigation sought to improve the handling and physical properties of a commonly used temporary zinc oxide-eugenol cement by changing the base/accelerator (B/A) ratio or combining it with a petroleum jelly or fluoride varnish. METHODS: Twelve modifications of a temporary cement were evaluated in terms of retentive strength, compressive strength at 24 h, film thickness and by scanning electron microscopy. RESULTS: Decreasing the B/A mixing ratio increased the retentive and compressive strengths, but reduced the film thickness of the cement. By increasing the percentage of incorporated petroleum jelly or fluoride varnish in the cement, there was a progressive decrease in the retentive and compressive strengths and in film thickness. CONCLUSIONS: Modifications of a zinc oxide-eugenol temporary cement to change the B/A ratio or to incorporate additives resulted in variations in physical properties. All modified forms of the cement had a film thickness less than 25 microns and a compressive strength below 35 MPa. With a wide range of retentive strength, modified forms of zinc oxide-eugenol cement may be found to have diverse clinical applications. PMID- 10666965 TI - Designing new treatment strategies in vital pulp therapy. AB - OBJECTIVES: The development of strategies in vital pulp therapy, which aim to maintain vitality and function of the dentine-pulp complex, represents a major focus of attention. Recent progress in understanding the molecular and cellular changes during tooth development and how they are mimicked during dental tissue repair offers the opportunity to now assess whether this knowledge can be exploited to design new treatment strategies in vital pulp therapy. DATA SOURCES AND STUDY SELECTION: Current literature on the molecular and cellular basis of tooth development and dental tissue repair has been reviewed in the context of stimulating dentinogenic responses in the tooth together with pertinent published abstracts of relevant conferences and personal communications. Tissue events of direct relevance to clinical application for vital pulp therapy are discussed. CONCLUSIONS: The involvement of growth factors and extracellular matrix molecules in signalling and regulating dentinogenic events during tooth development has been identified. During dental tissue repair, many of the processes are mimicked leading to responses of focal deposition of tertiary dentine at injury sites. The nature and specificity of these responses are determined in part by the extent of tissue injury. Traditional clinical strategies are capable of exploiting endogenous signalling molecules in the tissues to develop more effective treatment modalities. Application of exogenous signalling molecules offers opportunities for development of new therapies, although a number of delivery considerations must be addressed before these can be introduced into clinical practice. PMID- 10666966 TI - Subgingival calculus: where are we now? A comparative review. AB - OBJECTIVE: To critically analyse the formation, composition, ethnic variations and pathogenic potential of subgingival calculus in comparison with supragingival calculus. DATA SOURCES: Using CD-ROM and index medicus, scientific papers relating to subgingival calculus or subgingival and supragingival calculus written in the English language since 1960 were considered, with the emphasis on more recent articles. STUDY SELECTION: Studies were selected for their relevance and contemporary nature re:composition and formation of dental calculus and comparisons of ethnic groups with regard to dental calculus, especially subgingival calculus. Some similar studies were not included. DATA EXTRACTION: Abstracts of studies were kept brief unless particularly important to the review. Population, methodology, statistics and accurate conclusions were used as important guides to the quality and validity of studies. DATA SYNTHESIS: Similarities and differences between supragingival and subgingival calculus in composition and formation were shown. Different morphological types of subgingival calculus were demonstrated. There was evidence for an association between calculus formation and ethnicity with regard to supragingival and subgingival calculus, and an association between subgingival calculus composition and ethnicity was indicated. CONCLUSIONS: An association between ethnicity and subgingival calculus formation and composition was found. Further research into the reasons for these ethnic differences in dental calculus and the role of the mineral constituents especially of subgingival calculus would be valuable. PMID- 10666967 TI - Iatrogenic damage to approximal surfaces in contact with Class II restorations. AB - OBJECTIVES: This study investigated the frequency of iatrogenic damage to approximal surfaces in contact with Class II restorations. METHODS: Patients (n = 28) with a Class II restoration in contact with an unrestored surface had elastic separators fitted interproximally. Contralateral (unrestored) control surfaces were also separated. Impressions (light body polyvinylsiloxane) of the separated surfaces were taken 3-6 days later. Interproximal impressions (28 paired, seven unpaired) were examined by binocular microscope and scanning electron microscope (SEM) for iatrogenic damage, attrition and cavitation due to caries. RESULTS: 49 60% of surfaces adjacent to Class II restorations had been iatrogenically damaged. The most frequent types of damage were vertical grooves (26%), extensive damage (17%), indentations (6%) and scratches (6%). Damage was more frequent in maxillary teeth (61%) than mandibular teeth (25%), in permanent teeth (60%) more than deciduous teeth (20%). Qualified dentists produced more iatrogenic damage (64%) than undergraduate students (23%). CONCLUSION: The frequency of iatrogenic damage to approximal surfaces following Class II preparations was 49%, and possibly as much as 60% when questionably damaged surfaces were included. Protection of the adjacent enamel is of paramount importance during Class II cavity preparation. PMID- 10666968 TI - A five-year clinical evaluation of Class II composite resin restorations. AB - OBJECTIVES: To study the clinical efficacy of posterior composite resin restorations placed in general practice after five years. METHODS: Two commercial composite resin systems were used. Three general practitioners placed the restorations at a Public Dental Health Service Office. The patients were not selected specifically for this study. Class II cavities were restored with Superlux Molar and P-50 APC composite systems on an alternate basis. At baseline, 63 restorations were placed in molars and premolars in 45 patients. For primary caries, generally a conservative cavity design was used (n = 23), while replacements of amalgams resulted in the larger conventional Class II design (n = 40). The restorations were assessed using a modified USPHS criteria. Wear measurements were determined by the Leinfelder method. Photographs and bite-wing radiographs were taken to supplement the clinical evaluation of colour match, marginal adaptation and recurrent caries. Saliva sampling was performed to determine the rate of secretion and the level of mutans streptococci and lactobacilli. RESULTS: At the five-year review 51 restorations were available for examination, of which nine restorations were rated clinically unacceptable. Including the failed restorations (n = 7), at the three-year review, a total of 16 restorations had failed (27.6%) over a five-year period. The most common reasons for failure were recurrent caries (n = 7) and marginal defects (n = 4). The mean wear of Superlux Molar was 167 microns and of P-50 APC 158 microns. Eight of the 11 patients with failed restorations due to caries and marginal defects had high counts of mutans streptococci at baseline. CONCLUSIONS: The failures in the present group of patients did not specifically relate to material, tooth type or cavity design. However, it is suggested that patient factors such as caries activity should be monitored and managed. PMID- 10666969 TI - Three-year clinical evaluation of a polyacid-modified resin composite in minimally invasive occlusal cavities. AB - OBJECTIVE: The aim of this study was to evaluate the 3-year clinical performance of one polyacid-modified resin composite material (PMRC). Dyract, in minimally invasive occlusal cavities and its neighbouring fissures. METHODS: One hundred and sixteen restorations of the material investigated were placed by a single operator in a group of selected children under controlled conditions. Isolation of the restorations was accomplished with the use of cotton rolls and aspiration. Using modified US Public Health Service (USPHS) codes and criteria, the restorations were reviewed clinically within 1 week of placement (baseline), and thereafter at 6 months, 1, 2 and 3 years. RESULTS: After 3 years, marginal discolouration was present in 8.6% of the restorations. The marginal adaptation was rated as partly sealed (Oscar-Alpha) in 107 (92.2) of the restorations. Five restorations had lost their sealant components, while four restorations were partly sealed with explorer-catch after 3 years. Although wear of the restorations was considerable, restorations rated as 'partly sealed' had at least two-thirds of their sealant components fully retained. Recurrent caries was associated with four (3.4%) restorations. CONCLUSION: In this clinical study, the retention rate of the tested PMRC material was good, although a marked occlusal wear was evident. The marginal adaptation of the PMRC at the enamel site would probably have been better by the use of enamel-etching. Provided the marginal adaptation and wear resistance of the material is further improved, clinical use of PMRCs in minimally invasive occlusal cavities can be advocated. PMID- 10666970 TI - Clinical evaluation and microstructural analysis of a direct placement gallium restorative alloy. AB - OBJECTIVES: The objective of this study was to assess the clinical performance of a direct placement gallium alloy sealed with an established dentine adhesive system. In addition, microanalysis of a few gallium restorations that failed in clinical service was performed. Clinical factors such as pulpal sensitivity, fracture of the restoration and of the tooth, marginal deterioration, and tarnish were assessed. METHODS: Sixty-five restorations of Galloy and 62 of Tytin (49 and 51 Class II restorations, respectively) were placed according to a predetermined scheme for randomisation in 37 patients by two operators using rubber dam isolation. For the Galloy restorations, the enamel and dentine were etched, and then sealed with PAAMA 2 dentine adhesive according to the manufacturer's instructions. After carving, PAAMA 2 was applied to the Galloy and light-cured. Cavity preparations for Tytin received no adhesive sealer. All restorations were polished at least 24 h post-operatively. Microstructural analysis of retrieved fragments of failed restorations was conducted using electron probe microanalysis. RESULTS: At 1 year, only one Tytin restoration was found to have failed due to an isthmus fracture. The remaining restorations of Tytin were intact with no reported sensitivity. Of the 65 Galloy restorations placed, 28 had to be removed, including restorations in teeth, which were symptomatic, non-vital and/or fractured, and teeth with fractured restorations. Tarnish was present on many of the Galloy restorations. Retrieved fragments of failed Galloy restorations exhibited a dark surface at the pulpal wall interface and small cracks were observed in that surface. Internal cracks and extensive corrosion was observed using the microprobe. Gallium oxides and chlorides were identified as the predominant corrosion products. CONCLUSIONS: The gallium alloy, Galloy, sealed with PAAMA 2 dentine adhesive system demonstrated a high clinical failure rate. PMID- 10666971 TI - Accuracy of estimation of dental treatment need in special care patients. AB - OBJECTIVES: To assess the ability of carers and dental professionals to estimate treatment need in a group of children and adults with special needs. METHOD: A retrospective study of a series of 103 special needs patients who had received restorative dental treatment under general anaesthetic was undertaken. The initial reason for attendance and the time lapse between first symptoms and decision to consult were established. The parent or carer was asked to estimate treatment need and to assess the degree of discomfort suffered by the patient. The dentist evaluated treatment need by means of a pre-operative treatment plan. These results were compared to actual treatment performed. RESULTS: Treatment need was severely underestimated by both carer and dentist. The degree of advanced pathology found in the population would suggest that pain suffered was also underestimated. CONCLUSIONS: Access of patients with special needs to dental care may be limited by the ability of their carers to evaluate their oral condition and/or by the persons inability to express their pain or discomfort. PMID- 10666972 TI - Usage of denture adhesives. AB - OBJECTIVES: There have been few reports in the dental literature on the number of denture wearers who regularly use denture adhesive. The objectives of this study were to see what the incidence of usage of denture adhesive was by a surveyed edentulous population, to determine the degree of success of its usage and the reasons for its current use or nonuse. METHODS: In this study, 146 patients attending the Adelaide Dental Hospital for denture treatment were surveyed regarding their usage of denture adhesive, using a prepared questionnaire. The surveyed group was divided into three categories--those who had never tried denture adhesive, those who had tried denture adhesive but no longer used it, and those who currently used denture adhesive. RESULTS: In the survey, there were 52 males (35.6%) and 94 females (64.4%) and 96 (65.7%) were over 60 years of age. Of these 78 patients (54.9%) had worn their dentures for 10 years or more, and 26 (17.8%) for 20 years or more. A total of 98 patients (67.1%) had never tried denture adhesive: 48 patients (32.9%) had tried denture adhesive but only 10 (6.9%) currently used it. Various reasons for trying denture adhesive and for its continued use or nonuse were given. CONCLUSIONS: In this study, 52% of patients surveyed saw no need for the use of denture adhesive as they managed their dentures well; 20.5% of patients did not know that denture adhesives existed; and 32.9% had tried denture adhesive but only 6.9% continued to use it on a regular basis. PMID- 10666973 TI - A new imaging technique for measuring the surface strains applied to dentine. AB - OBJECTIVES: To investigate possible variation in directional material properties of dentine in relationship to tubule orientation using a new optical imaging technique. METHOD: The optical imaging technique records photometrically a grid pattern formed by using a transmission electron microscope grid as a template on the polished surface of the dentine. The grid pattern is silhouetted onto the sample surface using standard techniques. Compression (c) and diametral compression (dc) tests were undertaken using a servo hydraulic testing machine (MTS model 810) acting on rectangular blocks of dentine with dimensions 1.5 x 1.0 x 1.0 mm (for c) and cylindrical samples with dimensions 2.1 mm in diameter and 1 1.5 mm thick (for dc), respectively. The samples were cut using a diamond wheel and miniature lathe and the cut surfaces polished. Images due to a changing load profile were captured and stored as digitised files on a computer for later analysis. The precision is mainly determined by the pixel resolution of the charged-coupled device camera. RESULTS: Preliminary results show the value of elastic modulus of dentine (10.4 +/- 2.9 GPa) to be similar to those previously reported in the literature. Very small localised strains at the surface of a sample can be observed qualitatively and measured quantitatively by reference to the line spacing (approximately 85 microns). Maximum strength varied with tubule orientation and (compressive/tensile) stress. CONCLUSION: Very small samples of dentine may be investigated for strain in more than one direction using the imaging technique described. These results may be more appropriate for finding relative directional change rather than obtaining the elastic properties of the dentine. PMID- 10666974 TI - Effects of pH and concentration of citric, malic and lactic acids on enamel, in vitro. AB - OBJECTIVE: Dental erosion associated with soft drink consumption probably results from the contained dietary acids in the formulations. The pH value of any formulation is an important variable in acid erosion but not necessarily the only important factor. The aim of this study was to measure enamel erosion by citric, malic and lactic acids at pH values and acid concentrations representative of a range found in soft drink formulations and to determine the effect of adding calcium to citric acid. METHODS: Flat ground enamel samples were prepared from unerupted human third molar teeth. Groups of five specimens were placed in citric, malic and lactic acid solutions of different pH and acid concentration for three by 10 min exposures at 35 degrees C. Enamel loss was measured by profilometry. Enamel specimens were also exposed to citric acid solutions containing calcium at different pH values and at the same pH with different concentrations of calcium. RESULTS: Numerical data and contour plots for each acid showed a similar pattern for increasing erosion with decreasing pH and increasing acid concentration and vice versa for decreasing erosion. Increasing the concentration of calcium in a fixed pH citric acid solution resulted in decreased erosion. This effect was most marked at higher pH. CONCLUSIONS: This study has shown that under highly controlled conditions the erosion of enamel by solutions of dietary acids is influenced by the interplay of pH, acid concentration and presence of calcium. These variables and in particular the concentration of calcium could be manipulated to produce soft drinks with reduced erosivity to enamel. PMID- 10666975 TI - Accuracy and criteria for localizing arterial occlusion with transcranial Doppler. AB - The authors determined transcranial Doppler (TCD) accuracy for the proximal internal carotid artery (ICA), distal ICA, proximal middle cerebral artery (MCA), distal MCA, anterior cerebral artery (ACA), posterior cerebral artery (PCA), terminal vertebral artery (tVA), and basilar artery (BA) occlusion in cerebral ischemia patients. Detailed diagnostic criteria were prospectively applied for TCD interpretation independent of angiographic findings. Of 320 consecutive patients referred to the neurosonology service with symptoms of cerebral ischemia, 190 (59%) patients also underwent angiography (MRA or DSA). 48 of those 190 patients had angiographic occlusion and 12 of those 48 patients had involvement of multiple vessels. Median time from TCD until angiography was performed was 1 hour (41 patients had angiography before TCD). TCD showed 40 true positive, 8 false negative, 8 false positive, and 134 true negative studies with sensitivity 83.0%, specificity 94.4%, positive predictive value 83.0%, negative predictive value 94.4%, and accuracy 91.6% to determine all sites of occlusion. Sensitivity for each individual occlusion site was: proximal ICA 94%, distal ICA 81%, MCA 93% tVA 56%, BA 60%. Specificity ranged from 96% to 98%. TCD is sensitive and specific in determining the site of the arterial occlusion using detailed diagnostic criteria, including proximal ICA and distal MCA lesions. TCD has the highest accuracy for ICA and MCA occlusions. If the results of TCD are normal, there is at least a 94% chance that angiographic studies will be negative. PMID- 10666976 TI - Periventricular white matter hyperintensities on MRI: correlation with neuropathologic findings. AB - Periventricular white matter hyperintensities on postmortem magnetic resonance imaging (MRI) and myelin-stained frontal and parietal histologic sections were evaluated independently in 12 cases. There was a strong relationship between the extent of white matter hyperintensities on MRI and the extent of gross and microscopic changes seen in the white matter of myelin-stained sections, particularly in the frontal lobe. In this material, the extent of myelin rarefaction correlated with a 0- to 8-point white matter hyperintensity scale rating on MRI in the same brains. PMID- 10666977 TI - Color-coded duplex ultrasonography of the origin of the vertebral artery: normal values of flow velocities. AB - The introduction of color-coded duplex ultrasonography has improved the ease of performing ultrasound investigations of the vertebral arteries. So far, normal values of flow velocities have been reported only for the intertransverse region of the vertebral artery (V2 segments). Atherosclerotic disease at the origin of the vertebral arteries (V0 segment) is frequent and is one of the risk factors for vertebrobasilar ischemic disease. Normal values of flow velocities of the vertebral artery origin are needed to assess pathologic findings, such as vertebral artery origin stenosis or dissection. The aim of this study was to describe the normal flow velocities of vertebral artery origin (V0 segment) and the pre- (V1 segment) and intertransverse (V2 segment) part in 50 age-matched neurologic patients (mean age 54) without ischemic cerebral disease. The V0 segment could be visualized in 46 persons (92%) on the right side and in 43 (86%) on the left. The peak systolic blood velocity ranged from 30 to 100 cm/s (mean 63.6 +/- 17.5 cm/s), and end-diastolic blood velocity ranged from 10 to 35 cm/s (mean 16.1 +/- 5.1 cm/s). Analysis of side-to-side differences showed no significant differences of flow velocities in all subjects. It is concluded that color duplex ultrasonography is a feasible method to insonate the origin of the vertebral artery, and that nomogram data could be established. It is suggested that color-coded duplex ultrasonography of the vertebral artery origin should be performed in all patients with clinical symptoms or signs of vertebrobasilar ischemic disease. Nevertheless, further studies are needed to determine the normal and pathologic values of flow velocities of the vertebral artery origin and their reproducibility. PMID- 10666978 TI - Hyperkinetic movement disorders caused by corpus striatum infarcts: brain MRI/CT findings in three cases. AB - Three patients with hemichorea/hemiballismus/hemidystonia caused by discrete contralateral infarction of the corpus striatum are presented. The infarcts were all small on CT or MRI brain scan and were lacunar in type. Small discrete infarction of basal ganglionic structures allows such adventitious movements to be manifested. Involvement of contiguous areas, seen with larger infarcts, can suppress such movements. The infrequency of such hyperkinetic movement disorders, and the subtle infarct appearance on brain scan, can lead to a delay in the diagnosis. PMID- 10666979 TI - Intracranial clot dissolution is associated with embolic signals on transcranial Doppler. AB - Reperfusion of intracranial arteries can be detected by transcranial Doppler (TCD). The authors report microembolic signals (MES) on TCD as a sign of clot dissolution and recanalization. Microembolic signals were detected during routine diagnostic TCD examination performed in the emergency room in patients eligible for thrombolytic therapy. Microembolic signals were found at the site of M1 middle cerebral artery (MCA) high-grade stenosis or near-occlusion. Transcranial Doppler was performed before, during, and after thrombolytic therapy. Of 16 consecutive patients, 3 (19%) had MES on TCD. All three patients had a severe MCA syndrome at 2 hours after stroke onset scored using the National Institutes of Health Stroke Scale (NIHSS). In patient #1 (NIHSS 12), clusters of MES were detected distal to a high-grade M1 MCA stenosis preceding spontaneous clinical recovery by 2 minutes. Because of subsequent fluctuating clinical deficit, intraarterial thrombolysis was given with complete recovery. In patient #2 (NIHSS 20), TCD detected an M1 MCA near-occlusion. At 1.5 hours after intravenous tissue plasminogen activator, TCD showed minimal MCA flow signals followed by MES, increased velocities, and normal flow signals in just 2 minutes. She gradually recovered up to NIHSS 8 in 5 days. In patient #3 with NIHSS 22 and an M1 MCA near occlusion, TCD detected MES 15 minutes after TPA bolus followed by MCA flow velocity improvement from 15 cm/sec to 30 cm/sec. The patient recovered completely by the end of tissue plasminogen activator infusion. The authors conclude that embolic signals detected by TCD at the site of arterial obstruction can indicate clot dissolution. Intracranial recanalization on TCD can be associated with MES and changes in flow waveform, pulsatility, and velocity if insonation is performed at the site of arterial obstruction. PMID- 10666980 TI - Caffeine can affect velocity in the middle cerebral artery during hyperventilation, hypoventilation, and thinking: a transcranial Doppler study. AB - This study examined possible caffeine-mediated changes in blood flow velocity in the middle cerebral artery (VMCA) induced by tests of cerebrovascular responsiveness. Transcranial Doppler (TCD) sonography provided simultaneous bilateral VMCA measures while healthy college students hypoventilated, hyperventilated, and performed cognitive activities (short-term remembering, generating an autobiographical image, solving problems), each in 31-second tests. VMCA measures were obtained from the same persons, in separate testing sessions, when they were noncaffeinated and under two levels of caffeine: a smaller amount (from a cola, 45 mg/12 oz) and a larger amount (from coffee, 117 mg/8 oz). Compared with the no-caffeine control condition, a smaller amount of caffeine had no significant effects on global VMCA, but a larger amount suppressed VMCA by 5.8%. Time-course analyses showed that VMCA (1) followed a triphasic pattern to increase over baselines during hypoventilation regardless of caffeine condition, (2) slowed below baselines during hyperventilation (with the degree of slowing attenuated under caffeine), and (3) increased over baselines during all cognitive activities (ranges 3.8-6.9%). It is concluded that a large amount of caffeine can suppress VMCA, and this possibility should be anticipated when TCD is used to assess cerebral hemovelocity. PMID- 10666981 TI - Registration of neuroimaging data: implementation and clinical applications. AB - Image registration brings images into a form in which each voxel corresponds to a predetermined anatomic entity and is necessary for comparisons of data across scans. Intrasubject registration is a matter of translating and rotating one image volume into correspondence with another. Intersubject registration is more difficult because it requires the removal of individual anatomy dependence from the data. This article describes, with the help of clinical examples, automated methods for intrasubject registration of scans within and between modalities, and intersubject registration used for registering a three-dimensional brain atlas with a patient's brain scan. PMID- 10666982 TI - Basilar artery endoprosthesis placement: rescue therapy for recurrent thrombosis. AB - Hyperacute thrombosis of the basilar artery accompanied urgent treatment of basilar thrombosis with local thrombolytics and arterial reconstruction by balloon angioplasty. Successful placement of an endoprosthesis into the basilar artery permitted sustained restoration of blood flow. To the authors' knowledge, this is the first successful report of intracranial endoprosthesis deployment. PMID- 10666983 TI - The air we breathe. PMID- 10666984 TI - Health and the environment PMID- 10666985 TI - Water quality and public health in Georgia. PMID- 10666986 TI - Trash talk: burned and buried medical waste threatens Atlanta's environmental health. PMID- 10666987 TI - Expanding the physician's role in pediatric environmental health. AB - In rural Georgia, a nurse and an environmental health specialist from the local health department visit the home of a young nursing mother to evaluate her home for the presence of lead hazards. The mother's older child, a 3-year old girl, has a blood lead level of 22 micrograms per deciliter, which was discovered through routine (EPSTD) health department screening. In examining the home, the specialist finds classic environmental risks; peeling and chipping lead-based paint on windows and door frames, lead dust on window wells and floors, and a backyard that serves as a burial ground for defunct car parts and dead batteries. In the course of talking with the mother about the lead hazards he has found, he notices that she and her 8-week-old infant seem quite listless, so he asks the mother how she's coping with her new baby. Eventually, the mother discloses that during her pregnancy she craved dirt and that she had eaten bowels of it scooped from her backyard. Once she had the baby, she says, she lost her craving. The nurse immediately contacts the physician involved in this case, who arranges for the mother and infant to be admitted to a nearby medical center for chelation therapy. Testing reveals the mother's lead level at 90 micrograms per deciliter; the infant's level is staggering 85 micrograms per deciliter. Once lead levels are reduced,the physician and public health nurse arrange for a host of social services, including psychological and nutritional counseling for the mother and periodic retesting of the children. The family moves from the dilapidated rental home. However, the mother misses her appointments, and despite repeated attempts to locate her, the family is lost to follow-up. PMID- 10666988 TI - Air pollution and health: what Georgia physicians should know. PMID- 10666989 TI - Is universal screening for lead in children indicated? An analysis of lead results in Augusta, Georgia in 1997. AB - Pediatricians have a dilemma about whether they should do universal or targeted screening for lead in children. In order to determine whether we should continue our institution's present policy of universal screening, we analyzed 962 lead samples obtained by routine screening in the calendar year 1997, 83 children (8.6%) had elevated levels (over 10 mcg/dl) by capillary samples; of the 57 children who had follow up tests by venipuncture, 35 had abnormal levels making the incidence of elevated leads in our screened children 3.6%. Most of the abnormal levels were in African-American children living in two downtown zip codes. Based on recent AAP recommendations, in our community, targeted screening should replace universal screening. PMID- 10666990 TI - Endocrine disrupters: an emerging environmental health problem. PMID- 10666991 TI - What physicians can do for the environment. PMID- 10666992 TI - Exposure to contemporary-use pesticides. PMID- 10666993 TI - Monitoring the prevalence of congenital hypothyroidism for 20 years near the Savannah River site. AB - OBJECTIVES: The main objectives were to describe the epidemiology of primary congenital hypothyroidism (CH) in Georgia during the past 20 years and specifically to determine whether there was a significant increase in CH prevalence proximal to the Savannah River Site (SRS), a nuclear plant. METHODS: Data were derived from the Georgia Newborn Screening Program. Frequencies were determined for race, sex, birth weight and birth month. Seasonality effects were investigated and the prevalence was determined and mapped by health district. RESULTS: The majority of the CH cases were female, white and of average birth weight. The sex ratio varied by race/ethnicity. There was not a statistically significant increase in the prevalence in the eastern districts that were in close proximity to the SRS. CONCLUSIONS: Although there were differences in the prevalence between health districts, we found no increased prevalence of CH in those health districts proximal to the SRS. PMID- 10666994 TI - A Christian discovers the environmental crisis. PMID- 10666995 TI - Judaism and the environment. PMID- 10666996 TI - Focus on independence. PMID- 10666997 TI - An index of environmental burden: Georgia 1999. PMID- 10666998 TI - Healthy homes. PMID- 10666999 TI - New approaches to the detection and prevention of iron-deficiency anaemia. PMID- 10667000 TI - Current breastfeeding knowledge, attitude, and practices of mothers in five rural communities in the Savannah region of Nigeria. AB - The knowledge, attitude, and practices regarding breastfeeding of 310 mothers in five rural communities in Toto Local Government in Nassarawa State, Nigeria were investigated using a questionnaire. One hundred and sixty-two (52.3 per cent) mothers were illiterate while 148 (47.7 per cent) had either primary or secondary school education. Apart from giving babies colostrum, which was seen more amongst mothers with higher levels of education (p < 0.001), other practices investigated such as exclusive breastfeeding, demand feeding, 'rooming-in', and time of first breastfeed were not influenced by the mother's level of education. Fifty-four per cent of mothers did not give their babies colostrum. All mothers attended the antenatal clinic but only 103 (33.3 per cent) received instructions from the health worker on breastfeeding and 46.8 per cent delivered at home. Only 28.6 per cent of babies were breastfed within 24 hours of birth. The mean time after birth for the first breastfeed was 47.7 hours. Although breastfeeding is widely practiced, none of the babies was exclusively breastfed, and prelacteal feeds ranging from water, formula, or herbal tea were given by all the mothers. The practice of discarding colostrum and replacing it with a wide range of prelacteal feeds and late initiation of breastfeeding has implications for health education programmes and neonatal feeding strategies. PMID- 10667001 TI - Spontaneous and provoked growth hormone (GH) secretion and insulin-like growth factor I (IGF-I) concentration in patients with beta thalassaemia and delayed growth. AB - Growth retardation in children with thalassaemia major is multifactorial. We studied the growth hormone (GH) response to provocation by clonidine and glucagon, measured the circulating concentrations of insulin, insulin-like growth factor-I (IGF-I), IGF-binding protein-3 (IGFBP3), and ferritin, and evaluated the spontaneous nocturnal (12 h) GH secretion in prepubertal patients with thalassaemia and age-matched children with constitutional short stature (CSS) (height SDS < -2, but normal GH response to provocation). The anatomy of the hypothalamic pituitary area was studied in patients with abnormal GH secretion using MRI scanning. Children with thalassaemia had significantly lower peak GH response to provocation by clonidine and glucagon (8.8 +/- 2.3 micrograms/l and 8.2 +/- 3.1 micrograms/l respectively) than did controls (17.6 +/- 2.7 micrograms/l and 15.7 +/- 3.7 micrograms/l respectively). They had significantly decreased circulating concentrations of IGF-I and IGFBP3 (68.5 +/- 19 ng/ml and 1.22 +/- 0.27 mg/l respectively) compared to controls (153 +/- 42 ng/ml and 2.16 +/- 0.37 mg/l respectively). Seven of the thalassaemic children had a GH peak response of < 7 micrograms/l after provocation. Those with a normal GH response after provocation also had significantly lower IGF-I and IGFBP3 concentrations than controls. Analysis of their spontaneous nocturnal GH secretion revealed lower mean (2.9 +/- 1.77 micrograms/l) and integrated (2.53 +/- 1.6 micrograms/l) concentrations compared to controls (4.9 +/- 0.29 micrograms/l and 5.6 +/- 0.52 micrograms/l respectively). Five of them had mean nocturnal GH concentration < 2 micrograms/l and four had maximum nocturnal peak below 10 micrograms/l. These data denoted defective spontaneous GH secretion in some of these patients. MRI studies revealed complete empty sella (n = 2), marked diminution of the pituitary size (n = 4), thinning of the pituitary stalk (n = 3) with its posterior displacement (n = 2), and evidence of iron deposition in the pituitary gland and midbrain (n = 7) in those patients with defective GH secretion (n = 9). Serum ferritin concentration was correlated significantly with the circulating IGF-I (r = -0.47, p < 0.01) and IGFBP3 (r = -0.43, p < 0.01) concentrations. These data prove a high prevalence of defective GH secretion in thalassaemic children associated with structural abnormality of their pituitary gland. PMID- 10667002 TI - Role of intrauterine growth retardation on physical growth of Pakistani squatter children from birth to 2 years of age. AB - A birth cohort of 727 squatter children from Karachi was followed to study growth patterns by measuring anthropometric parameters at specific ages during the first 2 years of life. The mean weight and length of the intrauterine growth retarded and appropriate for gestational age children fell below the 10th percentile of the NCHS standards after 9 months and further deteriorated in the subsequent study period. However, the intrauterine growth retarded children showed slightly higher growth velocities compared to appropriate for gestational age children in the first few months for all four measurements, but subsequently these differences in growth velocities diminished. Our results suggest that nutrition intervention strategies should begin in early pregnancy. PMID- 10667003 TI - Comparison of four anthropometric methods of nutritional assessment and evaluation of the agreement between two reference populations. AB - A total of 841 children, aged 10 days to 5 years, seen at a primary health care centre in Ribeirao Preto, Brazil, were studied in order to compare the methods of Gomez, Waterlow, Shakir, and Kanawati and McLaren and to determine the correlation between the Brazilian and the National Center for Health Statistics (NCHS) reference populations, which are frequently used in Brazil. Anthropometric measurements (weight, height, arm circumference, and head circumference) were made and personal data were obtained in order to evaluate nutritional status according to the above methods. The two reference populations were compared by the method of Gomez. The NCHS and the Brazilian reference populations were concordant. Comparison of the methods of Gomez and Shakir showed a very mild agreement in the evaluation of nutritional status. Comparison of Waterlow 'wasting' with Kanawati & McLaren and with Gomez showed a mild agreement. PMID- 10667004 TI - Evaluation of the effect of a breastfeeding message integrated into a larger communication project. AB - Most breastfeeding project evaluations examine the effects of separate projects and many do not use experimental designs. We evaluated the impact during the 3rd year (March 1992-March 1993) of a simple breastfeeding message integrated into a large-scale 3 year vitamin A project in northern Bangladesh. It encouraged mothers to breastfeed for at least 2 years. Less formally, mothers were also advised to give colostrum. In both project (experimental) and non-project (control) areas, the proportion of mothers of children aged 1-6 years who said they gave colostrum increased from about 69 per cent to about 77 per cent. However, children born during the year that transpired between the pre- and post surveys were too young to be included in the post-survey. Thus this difference is an example of women reporting what they perceived as the norm or the 'correct' answer instead of what they actually did with their children. Thus the norm changed during the evaluation year, but it did so equally in both project and non project areas. Although reported breastfeeding levels remained stable among older groups, the proportion of children in the 2nd and 3rd years of life reported to be currently breastfeeding increased to a similar extent in both areas. This could indicate either a change in perception or a change in behaviour. Suckling frequency, not mentioned in local or national breastfeeding promotion, was unchanged in both areas at about 15 times a day for children in the 2nd year of life, and maintained at about eight to 10 times a day even for the small proportion who were still breastfed in the 5th (about 9 per cent) or 6th (about 2 per cent) years of life. While project messages (especially by radio) could have spread to the nearby non-project area, it is unlikely that the impact would be equal. More likely these changes were the result of secular changes occurring at this time throughout Bangladesh. Thus this evaluation found no evidence that the integration of a simple message about breastfeeding into a larger project had any measurable effect in the project area compared to the non-project area. More effective approaches may be required, tailored to the particular needs of women in the area. PMID- 10667005 TI - Fatal mumps nephritis and myocarditis. AB - The case of a 14-year-old girl with fatal interstitial nephritis and myocarditis as complications of mumps is reported. The illness began with parotitis; renal symptoms developed within a week. The patient's renal and cardiac status and clinical course rapidly deteriorated and the outcome was fatal. The post-mortem renal biopsy sample showed interstitial mononuclear cell infiltration, oedema, and focal tubular epithelial damage in biopsy material of kidney, confirming the clinical diagnosis. Myocarditis was determined by electrocardiographic and echocardiographic findings. Since it has been reported that fatal complications such as myocarditis, dilated cardiomyopathy, and nephritis may develop in the course of mumps, the patients with mumps, especially in complicated cases, should be followed closely because of the severe clinical conditions which may progress. PMID- 10667006 TI - A child with Neisseria meningitidis endocarditis and Staphylococcus aureus septicaemia. PMID- 10667007 TI - Paediatric intensive care in Kuala Lumpur, Malaysia: a developing subspecialty. AB - Paediatric intensive care in Malaysia is a developing subspecialty with an increasing number of specialists with a paediatric background being involved in the care of critically ill children. A part prospective and part retrospective review of 118 consecutive non-neonatal ventilated patients in University Hospital, Kuala Lumpur was carried out from 1 June 1995 to 31 December 1996 to study the clinical epidemiology and outcome in our paediatric intensive case unit (PICU). The mean age of the patients was 33.9 +/- 6.0 months (median 16 months). The main mode of admission was emergency (96.6 per cent) with an overall mortality rate of 42 per cent (50/118). The mean paediatric risk of mortality (PRISM) score was 20 +/- 0.98 SEM, with 53 per cent of patients having a score of over 30 per cent. Multiorgan dysfunction (MODS) was identified in 71 per cent of patients. Admission efficiency (mortality risk > 1 per cent) was 97 per cent. Standardized mortality rate using PRISM was an acceptable 1.06. The main diagnostic categories were respiratory (32 per cent), neurology (22 per cent), haematology-oncology (18 per cent); the aetiology of dysfunction was mainly infective. Non-survivors were older (29.5 vs. 13.8 months, p < 0.0001), had more severe illness (mean PRISM score 30 vs. 14, p < 0.0001), were more likely to develop MODS (96 vs. 53 per cent, p < 0.0001) and required more intervention and monitoring. Paediatric intensive care in Malaysia differs widely from that in developed countries in patient characteristics, severity of illness, and care modalities provided. PMID- 10667008 TI - Perinatal mortality in twin deliveries in a general hospital in Zambia. AB - Data from birth records and death certificates were used retrospectively to determine the risk factors associated with perinatal mortality in twin pregnancy at Mansa General Hospital between 1 August 1993 and 31 July 1995. A total of 3091 singleton births, 128 sets of twins, and three sets of triplets was recorded during the study period with a twinning rate of 4 per cent of the total deliveries. Perinatal mortality in twin gestation was high (141/1000) and affected mostly the second twin (72 per cent). The main risk factors associated with perinatal mortality in twin pregnancy were birthweight less than 2000 g (47 per cent), retained second twin (36 per cent), primiparity (28 per cent), fetal malpresentations (25 per cent), cord anomalies (17 per cent), and home deliveries (14 per cent). Perinatal deaths in twin pregnancies can be reduced by early recognition of twin gestation during antenatal visits, prevention of premature labour, and provision for emergency referral. A woman carrying twins should be advised to deliver in a well equipped health facility, with adequately trained health personnel rather than the traditional midwife, in order to minimize associated hazards. PMID- 10667009 TI - Survival of childhood acute lymphoblastic leukemia: experience in Chennai. AB - The object of this study was to evaluate the treatment outcome in children with acute lymphoblastic leukemia (ALL) in Chennai. The problems inherent in a developing country which affect outcome are analyzed. The importance of prognostic factors especially immunotyping is assessed. The period of study was from June 1991 to December 1995. A total of 135 children were studied. Pre B CALLA positive (CD10, CD19, HLA, DR) was the dominant immunotype in 75 children (69 per cent). T-cell ALL was seen in 15 (14 per cent), biphenotype in three (2 per cent), and B in one (0.9 per cent). Seventy children (53 per cent) were treated with a high risk protocol, 25 (17 per cent) received an intermediate risk, and 40 patients (30 per cent) received a standard risk protocol. Analyzing the outcome in 135 children, 34 (27 per cent) had event free survival (EFS) at the time of analysis; of these 41 per cent had EFS after 2 years of therapy, 31 per cent after 3 years and 18.7 per cent after 4 years (i.e. 1 year after stopping 3 years of therapy). Fifty-seven children (41 per cent) dropped out; 25 (18 per cent) died due to sepsis. Treatment obstacles included delay in diagnosis, poor health education and facilities, poor supportive care, and socio economic problems. PMID- 10667010 TI - Alpha thalassaemia in Yemeni children with sickle cell disease. AB - Alpha thalassaemia frequently occurs in several of the Middle Eastern populations. This study was conducted on 26 sickle cell disease (SCD) patients from Yemen and 19 normal children (Hb AA) living in Riyadh, Saudi Arabia. Blood samples were extracted by venepuncture, and haematological and biochemical parameters were estimated. DNA was extracted from the buffy coat and analysed for alpha-gene arrangement using Bam HI and Bgl II. The frequency of alpha-gene deletion in the total Yemeni group (26 SCD + 19 Hb AA) was 0.311 for one alpha gene deletion (-alpha/alpha alpha) and 0.13 for two alpha-gene deletions (-alpha/ alpha). When separated on the basis of the Hb phenotype the alpha-gene deletion frequency was significantly higher (-alpha/alpha alpha = 0.346 and -alpha/-alpha = 0.231) in the SCD patients compared to the normal Hb AA group (-alpha/alpha alpha = 0.263 and -alpha/-alpha = 0). In the Hb AA group one child had triple alpha-gene arrangement (alpha alpha alpha/alpha alpha) giving an overall frequency of triple alpha-gene as 0.022. Haematological parameters showed variations in the SCD patients with and without alpha-gene deletion. This paper shows for the first time that alpha-gene deletion occurs in the Yemenis and the frequency is higher in patients with SCD. Further population-based studies are required to determine the exact frequency of the different types of alpha thalassaemias in the overall Yemeni population. PMID- 10667011 TI - Thyroid hormone studies in protein-energy malnutrition. PMID- 10667012 TI - Serum vitamin A levels in children with persistent diarrhea and impact on clinical profile. PMID- 10667013 TI - Fine needle aspiration cytology in the management of childhood palpable masses: Ibadan experience. PMID- 10667014 TI - The health status of rural school children in the Amazon basin of Ecuador. AB - An assessment of the health status among school children of the Naporuna ethnicity was conducted in north-eastern Ecuador. Prevalence of protein-energy malnutrition (PEM), parasitic infections, and pathology was investigated among 511 school children. The overall nutritional status of the school children was found to be good despite a high prevalence of helminthic infections. The prevalence of stunting was 1.4 per cent and of wasting 1.8 per cent. Sixty-four per cent of the children surveyed were found infected with one or more of the soil-transmitted helminths. In the clinical examination high prevalence of pterigium (89.2 per cent) was found. Upper respiratory infections (5.2 per cent), septic skin lesions (4.4 per cent), mycotic otitis (3.8 per cent), tinea (3.4 per cent) and bronchitis (2 per cent) were the main infectious pathology found. Likely explanations of these findings are discussed. PMID- 10667015 TI - [Surgical treatment for bilateral multiple lung cancers]. AB - We reviewed 12 patients with contralateral bronchogenic carcinomas. Seven of them had metachoronous carcinomas and 5 had synchronous carcinomas. We treated 3 patients with lobectomy on both lungs, and 4 patients with lobectomy and segmentectomy, 2 patients with lobectomy and wedge resection, 2 patients with segmentectomy and thoracoscopic wedge resection, and one patient with lobectomy and ablation on each lung. Two patients who had lobectomy on both lungs were dead, one of whom of bronchofistula on operation and the other of respiratory failure 7 years and one month after second operation. The 5-year survival rate in 12 patients was 68.5% after first operation and 82.5% after second operation. We conclude that lobectomy on both lungs are not recommended because of high mortality rate and the limited resection should be considered to treat the other contralateral primary lung cancers. Because the patients with primary lung cancers have the possibility to suffer from new primary cancers in the different site of the lung, we need careful follow up of the patients after treatment on the first lung cancer. PMID- 10667016 TI - [A successful case of hybrid therapy for the left main trunk and triple coronary vessel lesions with acute myocardial infarction and cardiogenic shock]. AB - A 86-year-patient who had acute myocardial infarction and critical cardiogenic shock was diagnosed to have the left main trunk (LMT) and triple vessel disease. Emergent coronary artery bypass grafting to the left anterior descending artery was performed using saphenous vein graft without cardiopulmonary bypass through median sternotomy. On the 41st postoperative day, catheter intervention was performed to the remaining lesions by stenting of LMT and percutaneous transluminal coronary angioplasty to the right coronary artery lesions. Tl scintigraphy showed remarkable reduction of myocardial ischemia. Hybrid therapy is the effective new strategy for critical cases which cannot be successfully and securely treated by medical or surgical approach alone. PMID- 10667017 TI - [Synchronous primary triple cancers including the lung, stomach, and thyroid: a case report]. AB - A 62-year-old man with synchronous multiple primary cancers involving the lung, stomach, and thyroid was admitted. Initially the patient's chest X-ray showed an abnormal shadow in the right middle-lobe indicating lung cancer. During preoperative examination, gastric cancer of the antrum and angle were detected. Excisional biopsy of the lymph node in the neck after chest surgery revealed thyroid cancer. A middle lobectomy with mediastinal lymph node dissection was performed for lung cancer and the histological diagnosis was moderately differentiated adenocarcinoma, pT4N2M0, stage IIIB. Gastric cancer was treated by endoscopic mucosal resection. Considering the relatively better prognosis of papillary thyroid cancer, we concluded that no further treatment to the thyroid lesion was necessary. In Japan, according to autopsy reports, triple primary cancers are gradually increasing. During the periods 1994 to 1996, the incidence of triple cancers was 0.81% of all autopsy cases reported. PMID- 10667018 TI - [Transaortic video-assisted thrombectomy in the left ventricular cavity after acute myocardial infarction]. AB - In recent years much attention has been paid to a minimal invasive surgery even in the field of cardiac surgery. We have successfully treated a 59 year-old male who underwent transaortic video-assisted thrombectomy in the left ventricular cavity following acute myocardial infarction due to occlusion (No. 7) of the left descending coronary artery (LAD). After the LAD was recanalized by percutaneous transluminal coronary angioplasty (PTCA) and then stenting, this thrombectomy was carried out. Following initiation of cardiopulmonary support, thrombus in the left ventricular apex was safely removed by transaortic endoscopic guidance. This technique was useful especially for the patient with poor ventricular function due to acute myocardial infarction because of no left ventriculotomy. Endoscopic guidance technique is reported in detail. PMID- 10667020 TI - [Clinical experience using percutaneous cardiopulmonary support system for stent replacement]. AB - Three cases of tracheal or mein bronchus stenoses were treated using percutaneous cardiopulmonary support system (PCPS). Case 1 was a 63-year-old male admitted for dyspnea due to stenotic trachea with primary lung cancer invasion. YAG-laser operation and Dynamic stent was inserted to the trachea using PCPS. Case 2 was a 74-year-old male admitted for dyspnea due to stenotic right mein bronchus with primary lung cancer invasion. Dumon Y stent was inserted to the right mein bronchus using PCPS. Case 3 was 57-year-old male admitted for dyspnea due to stenotic trachea and occluded left mein bronchus with ischemic change after primary esophageal cancer operation. Dynamic stent was inserted to the trachea and left mein bronchus using PCPS. Tracheal and mein bronchus stenoses the trachea of all was dilated after placement of stent. These three cases had no complications during or after these treatment. These results indicated that using PCPS was a very useful, powerful and satisfactory method in the treatment of tracheal or mein bronchus stenoses during the lack of lung ventilation. PMID- 10667019 TI - [Coronary artery bypass grafting for a patient with bronchial asthma seceeded from cardiopulmonary bypass by additional bypass for spasm of radial artery graft: a case report]. AB - A 78-year-old male who had a bronchial asthma underwent coronary artery bypass grafting (CABG) using the left internal thoracic artery and the radial artery. The patient could not be weaned from the cardiopulmonary bypass because the radial artery which anastomosed to the obtuse marginal artery (OM) had a spasm after CABG. An additional bypass using a long saphenous vein to OM was carried out immediately. It brought a weaning from cardiopulmonary bypass. If the cardiac function after CABG is insufficient in patients with bronchial asthma, CABG must be re-done immediately, considering that they cause the arterial spasm more than patients without bronchial asthma. PMID- 10667021 TI - [The effects of intravenous milrinone for the patient undergoing CABG]. AB - The hemodynamic effects of intravenous infusion of milrinone were evaluated in 25 patients undergoing CABG using an internal mammary artery graft. Milrinone was administered to 9 patients at the time of weaning from cardiopulmonary bypass, at a dosage of 3 to 5 micrograms/kg/min. Postoperative cardiac function was compared in this group versus the other 17 patients who were treated without milrinone. We determined such parameters as cardiac index, wedge pressure and mean pulmonary pressure. Our findings did not show any significant difference between the 2 groups. We also studied a subject of low-output patients (EF < 0.5). In the patients with low-cardiac output, the use of milrinone in addition to standard postoperative administration of low-dose dopamine reduced mean pulmonary pressure and wedge pressure. Thus, milrinone not only improved the left ventricular function, but also expanded the pulmonary vascular bed. PMID- 10667022 TI - [Dose the serum brain natriuretic peptide (BNP) level after open heart surgery reflect myocardial protection?]. AB - This clinical study was conducted to determine whether the serum BNP level after open heart surgery reflects myocardial protection. The levels of BNP and CPK-MB were measured before and after 12 hours of cardiopulmonary bypass, then 1, 3, and 6 days after open heart surgery, and the relationship between the maximum levels of BNP and the CPK-MB after open heart surgery was examined. The patients were divided into two groups according to whether or not the maximum CPK MB was more than 100 IU/l after open heart surgery. A significant relationship between the maximum BNP and the maximum CPK-MB after open heart surgery was observed (p = 0.013). Moreover, the BNP was significantly increased in the group of patients with a maximum CPK-MB > or = 100 IU/l, compared to that in those with a maximum CPK-MB < 100 IU/l, 12 hours 1 day, and 6 days after open heart surgery (p < 0.01). These findings indicate that the serum level of BNP after open heart surgery can reflect myocardial protection. PMID- 10667023 TI - [Surgery for intrathoracic recurrence of non-small cell lung cancer]. AB - In this study, we defined a solitary lung nodle in the same histology which could be traced its' origin from carcinoma in situ or was found over than two years' follow up as a second primary lung cancer. These cases were excluded. Eighteen cases underwent second surgery for intrathoracic recurrence. Fourteen cases were male and four cases were female. Their ages ranged from 23 to 75 (average 59.6) years. The histology were adenocarcinoma in 9 cases, squamouscarcinoma in 7, adenosquamous carcinoma in 1, large cell carcinoma in 1. The initial surgical procedures were lobectomy in 17, partial resection in 1. The initial stage were I in 13, II in 2, IIIA in 1. Pulmonary recurrence were found in 10, bronchial stump recurrence were found in 4, pulmonary hilus lymph node recurrence were found in 2, mediastinal lymph node recurrence were found in 2, pulmonary stump recurrence was found in 1. The second surgical procedures were completion pneumonectomy in 7, completion lobectomy in 1, lobectomy with segmentectomy in 1, segmentectomy or partial resection in 7, mediastinal dissection in 2. The overall 5-year survival rate of the patients with recurrence after reoperation was 31.8%. An aggressive surgical approach for recurrent lung cancer should be recommended. PMID- 10667024 TI - [Late results of SJM and CM valves in bentall procedure]. AB - Recently, Bentall procedure is commonly performed for annuloaortic ectasia with aortic regurgitation or dissecting aneurysm. And the operative results are improving. In this study, we evaluated results of the St. Jude Medical (SJM) and Carbomedics (CM) valves which were used in this procedure. From 1979 to 1994, 87 SJM valves and 22 CM valves were implanted in the aortic position of Bentall procedure. Total follow-up was 528.6 years in the SJM group and 56.5 years in the CM group. According to the Kaplan-Meier actuarial method and the Cox-Mantel statistical analysis, actuarial survival, thromboembolism free rate, reoperation free rate, event free rate were not different between the SJM and CM groups. These results suggest that, current selection of the SJM and CM valves would be acceptable in Bentall procedure. PMID- 10667025 TI - [Ineffective and recurrent cases of thoracoscopic sympathectomy for hyperhidrosis and intractable pain]. AB - We reported the cases of thoracoscopic sympathectomy, that is, six cases of hyperhidrosis, three of post herpetic neuralgia, and four of reflex sympathetic dystrophy, including recurrent or incompletely resected or ineffective ones. Recently this procedure for hyperhidrosis had been performed frequently because of its effectiveness, less pain, early discharge and cosmetic aspect. For an ineffective case of hyperhidrosis abdominal respiration which emphasized the exhalation and using an upper abdomen decreased the sweating. The balance of autonomic nerve system, toward parasympathetic dominant, was thought to be improved by conscious respiration. The decrease of sweating right after the operation in a case of incomplete resection indicated that intraoperative maneuver could restrict the sympathetic nerve. This procedure for a pain control could be less effective than that for hyperhidrosis, so an adequate preoperative informed consent was thought to be necessary. PMID- 10667026 TI - [An elderly case of ruptured aortic arch aneurysm with hemorrhagic cardiac tamponade]. AB - We reported a case of successful aortic arch replacement using selective cerebral perfusion for ruptured distal aortic arch aneurysm (DAAA) with cardiac tamponade. A 80-year-old man who had preoperative episode of severe chest pain. Computed tomography showed saccular DAAA and pericardial effusion. He was diagnosed as ruptured DAAA with hemorrhagic cardiac tamponade. We performed urgent graft replacement of the aortic arch using selective cerebral perfusion. Postoperatively he had no complication. Thirty days after the operation he was discharged from the hospital and he is now leading a normal life. PMID- 10667027 TI - [Blunt rupture of the intrapericardial inferior vena cava: report of two cases]. AB - Blunt rupture of the intrapericardial inferior vena cava is rare. Our experience in recent two cases is presented. Case 1: A 52-year-old male was admitted following a traffic accident. Chest CT demonstrated cardiac tamponade and mediastinal hematoma. Ruptures of the right and left atria across the caudal aspect of the atrial septum, and a separate laceration of intrapericardial IVC were found in the emergency operation. Case 2: A 35-year-old male jumped from the fourth floor of a building. Chest CT revealed descending aortic rupture and the patient was taken to surgery. He died of massive hemorrhage from the aortic rupture. Exploration revealed a rupture of intrapericardial IVC. Recent literatures were reviewed and the mechanism of IVC rupture is discussed. PMID- 10667028 TI - [Ventricular septal defect with pulmonary hypertension and concomitant left atrial myxoma in elder patient: a successful surgical case report]. AB - A 70-year-old man was admitted with syncope attack on exertion. Ventricular septal defect with pulmonary hypertension and left atrial myxoma were confirmed by ultrasonography and cardiac catheterization. Preoperative Pp/Ps was 0.95 and pulmonary vascular resistance was 16 units. Pulmonary vascular resistance decreased to 9.6 units by the administration of Isoproterenol and decreased to 8.5 units with PGE1. Patch closure of VSD and excision of left atrial myxoma were performed simultaneously. The patient recovered completely, although he suffered from pneumonia and jaundice due to liver congestion postoperatively. Cardiac catheterization before discharge revealed Pp/Ps 0.38 and PVR 10.1 units. PMID- 10667029 TI - [A case report of primary hemangiopericytoma of the chest wall]. AB - A 35-year-male was found to have an abnormal shadow with an extrapleural sign located in the right lower lung field by a chest X-ray. Chest CT revealed a well demarcated tumor in the chest wall adjacent to the 4th rib. Chest MRI showed that the tumor contained punctate or linear low-intensity areas, which were considered to be small blood vessels. A diagnosis of hemangiopericytoma was established by percutaneous needle biopsy. Under the definite diagnosis, extended resection of the chest wall was performed to remove the tumor with a surgical margin of more than 5 cm, corresponding to surgery for other malignant soft-tissue neoplasmas. Hemangiopericytomas rarely arise in the chest wall, although they can be found in any region which contains pericytes. Preoperative definite diagnoses of hemangiopericytoma have rarely been reported. However, preoperative diagnosis is an important factor in deciding the operative procedure for hemangiopericytoma. It has been reported that hemangiopericytomas show local recurrences and distant metastases, although they are histologically benign. We consider that hemangiopericytomas in the chest wall should be treated with extensive resection corresponding to surgery for other malignant soft-tissue neoplasmas. PMID- 10667030 TI - [A case of mediastinal mature teratoma accompanying elevated CA 19-9 and CEA in the cystic fluid]. AB - Fourteen year old female visited our hospital suffering from anterior chest pain. Chest CT revealed anterior mediastinal mass which had multiple cystic lesions, 8.5 x 7.0 x 5.5 cm in size. Tumor extirpation had performed by median sternotomy and pathological diagnosis was mature teratoma. CA 19-9 and CEA in the cystic fluid had elevated 32,790 U/ml and 768 ng/ml, respectively. PMID- 10667031 TI - [A case of teratoma in both the mediastinum and the intrapulmonary system]. AB - A rare case of mature teratoma in both the mediastinum and the intrapulmonary system is presented. A 30-year-old male was admitted to our hospital due to tumor masses in the mediastinum and the left lung. We performed mediastinal tumor resection and left upper partial lobectomy. Neither tumor communicated with each other. Pathological findings revealed teratoma in the mediastinal lymph node and the intrapulmonary system including no malignant cells in either tumor. In this case, because metastasis and perforation were negative, we proposed that both tumors occurred at the same time in the early embryo. PMID- 10667032 TI - [Postoperative interstitial pneumonia: 2 cases after lobectomy]. AB - Two patients with postoperative interstitial pneumonia are reported. Preoperative diagnosis was primary lung cancer without idiopathic interstitial pneumonia (IIP). Within one week after operation acute interstitial pneumonia (AIP) occurred on the nonoperated side and developed. Steroid therapy was performed but one was dead. AIP is a fatal complication after pulmonary resection and steroid therapy may be useful in some cases of postoperative AIP. PMID- 10667033 TI - Perioperative assessment; what is the question? PMID- 10667034 TI - Preoperative screening and preoperative medicine: a new challenge for anesthesiology and internal medicine. PMID- 10667035 TI - The value of peri-operative consultation on a general surgical ward by the internist. AB - BACKGROUND: Medical peri-operative consultation plays an important role in the practice of the internist. It represents 13-33% of the total consultation done by the internist. The value of preoperative consultation by the internist is still unclear and the place of the consultations is under discussion. METHODS: Consecutive medical consultations--by internist-intensivists from the surgical ICU--of 408 patients in the department of Surgery of the University Hospital Groningen were retrospectively analyzed, with specific attention to the consultation request, the consultation report and the outcome of the consultation. RESULTS: The main problems were cardiac (34%) or pulmonary (20%). In 78% well defined questions about the patients were asked and in 29% of the preoperative requests, the requesting surgeon asked specific advice about diagnosis and management. In 49% the requesting physician asked for an 'evaluation' of the patients. Of all consultations 271 (67%) were preoperative consultations. In 12% the findings of the preoperative consultation had a significant impact on the course of the patient, whereas in 7% of the preoperative requests the operation was postponed. The consulting internist recommended in 2% to cancel the operation for this admission, which was shared by the surgeon and anesthesiologist. In 10% of all cases new diseases or processes were diagnosed, or were found not to be treated adequately before consultation, so the diagnosis or management was changed. CONCLUSIONS: Our data show that (semi ) elective consultations by internists at the department of Surgery, on indication of the surgeon, changes the course of a significant percentage of patients. (See Editorials p. 1 and p. 4). PMID- 10667036 TI - Clinical and pharmacological aspects of accidental triamcinolone acetonide overdosage: a case study. AB - Local administration of corticosteroids for rheumatic diseases have had a long history of effective and well-tolerated use. We report here the pharmacodynamics and pharmacokinetics of an accidental triamcinolone acetonide (TCA) overdose. The presented patient was treated with 200 mg TCA and developed Cushing's syndrome 6 weeks later (cortisol and ACTH concentrations were below limits of detection, TCA concentrations were > 3 micrograms/l). Because of her severe symptoms, mifepristone was administered for a period of 19 days. Cortisol concentrations became detectable 2 days after initiation of mifepristone treatment and persisted, being detectable for a period of at least a week after cessation of the drug. Twenty days after cessation, cortisol concentrations were undetectable again. Cushing's syndrome persisted more than 6 months while TCA concentrations remained detectable for at least 80 days. Based on plasma TCA concentrations in our patient, we calculated a terminal half-life of TCA of 33 days as opposed to 5 days observed after intra-articular administration of a therapeutic dose of 40 mg TCA. We conclude that after an accidental overdose in this patient, body TCA disappearance was strongly prolonged due to a very slow (absorption) half-life of the drug in comparison to a therapeutic dose. This finding is explained by a 'flap-flop phenomenon' where drug absorption is the rate-limiting step of overall drug disposition. Caution is, therefore, needed to prevent undesired accumulation of TCA that may lead to protracted Cushing's syndrome. PMID- 10667037 TI - Inflammatory pseudotumor of the fossa pterygopalatina: diagnosis and treatment. AB - Pseudotumor is a term used to describe a space-occupying inflammatory lesion of unknown etiology that clinically simulates a neoplastic process. Pseudotumors of the fossa pterygopalatina and fossa infratemporalis are very rare. In this paper, we describe a patient who developed a pseudotumor in the left fossa pterygopalatina, secondary to an unclassified autoimmune disease, which caused progressive left-sided facialdynia and swelling. The tumor was detected with somatostatin receptor scintigraphy. The lesion was refractory to steroids, also in combination with azathioprine, as well as to surgical intervention. An excellent clinical response was observed after cyclosporine was added. This case is presented here in order to draw attention to the use of somatostatin receptor scintigraphy as a diagnostic tool in visualizing pseudotumors and to document a case that responded excellently to treatment with a combination of low-dose cyclosporine and steroids. PMID- 10667038 TI - A homozygous M694V mutation of the MEFV gene in a patient with periodic fever and thoracic pain. AB - A Turkish patient with episodic fever and thoracic pain is described in whom a homozygous M694V mutation of the MEFV gene confirmed the clinical diagnosis of familial Mediterranean fever. The role of DNA analysis is discussed with respect to understanding the pathogenesis of the fever and assessing the risk of amyloidosis in specific mutations of the MEFV gene. PMID- 10667039 TI - Chylothorax. AB - Chylothorax is defined as an accumulation of chyle in the pleural space caused by disruption of the thoracic duct or one of its major divisions. Chyle has a high content of triglycerides. The odorless fluid is turbid and milky due to the presence of fat containing particles, the chylomicrons. The etiology of chylothorax can be divided into four major categories: tumor, trauma, idiopathic and miscellaneous. Although chylothorax is uncommon, it is a serious and potentially hazardous disorder. Loss of chyle leads to metabolic disturbances, malnutrition and immunodeficiency. Treatment consists of treatment of the underlying disease, conservative treatment (medium chain triglyceride diet, parenteral nutrition) or surgical intervention. Appropriate timing of surgical intervention is essential. Since the ligation of the thoracic duct can be performed during thoracoscopy, this minimal interventional technique is the procedure of choice when conservative treatment fails. PMID- 10667040 TI - [Image of the month. Isolated appendicular polyposis]. PMID- 10667041 TI - [How I treat ... idiopathic hyperhidrosis]. AB - Palmo-plantar and/or axillary idiopathic hyperhidrosis is unpleasant and sometimes invalidating. Its malodorous variant which is due to biodegradation by bacterias is called bromhidrosis. According to the severity of the condition, several therapies exist including psychotherapy, various topical products, iontophoresis, botulinic toxin injections and thoracic sympathectomy. PMID- 10667042 TI - [Clinical case of the month. Cerebral salt wasting syndrome: report of a case]. AB - A 75 year old woman was found to have a posterior cerebellar lesion which after surgical removal was shown to be a meningioma. Her postoperative course was complicated by a MRSA meningitis and on day 23 after resection, a polyuria up to 11.7 1/24 h became apparent. The diagnosis of cerebral wasting syndrome (CSWS) was made based on biological and clinical features such as an excessive natriuresis (143 mmol/l) resulting in hyponatremia (130 mmol/l) and an osmolarity higher in urine than in blood. A low central venous pressure and a low wedge pressure confirming a volumic depletion indicated the diagnosis of CSWS. This syndrome has marked similarities with the Inappropriate Secretion of Antidiuretic Hormone Syndrome (SIADH) in terms of biological finding with regards to clinical context and presentation. Without an adequate assessment, a patient with CSWS may be misdiagnosed as SIADH. However recognition is important, as water restriction which is part of SIADH treatment, is detrimental to patients with CSWS and can possibly be lethal. PMID- 10667043 TI - [Medication compliance]. AB - Compliance is defined as the extent to which a patient's behaviour coincides with medical or health advice. Medication compliance seems to be rather low as 30 to 60% of no or poor adherence to medical recommendation have been reported. Numerous factors may influence medication compliance among which patient's characteristics, disease particularities, drug treatment modalities or physician's attitudes. The consequences of medication non-compliance may not only be dangerous for patient's health, but also dramatically increase the financial cost for public health services. Thus, all energies should be devoted to improve drug compliance, including treatment optimization and simplification, patient's information and education, use of practical means that facilitate adherence to medical recommendation, the patient being responsible for his/her treatment. PMID- 10667044 TI - [Anatomic-clinica dilemma. Case report of renal cell carcinoma]. AB - The authors report the case of a patient with a history of hypertension and multiple intracerebral hemorrhages who was found at post mortem examination to have a renal cell carcinoma. The relationship between renal carcinoma and hypertensive intracerebral hemorrhages is discussed. PMID- 10667045 TI - [Winter skin diseases]. AB - Winter climate is prone to induce cutaneous and systemic alterations mediated by cold. Xerosis due to the impairment of the desquamation process is not rare on the limbs. Chilbain results from reversible alterations of the dermal vasculature. Cold panniculitis is the consequence of lipid crystallization within adipocytes. More dramatic issues occur when cold exposure extends beyond skin. They are represented by frostbite and body hypothermia. Intensity and duration of cold exposure combined with wind speed, altitude and environmental hygrometric value govern the potential type of low temperature-dependent pathology. Ultraviolet irradiation can also induce cutaneous lesions during winter time. PMID- 10667046 TI - [New players in the physiopathology of water metabolism: the aquaporins]. AB - Aquaporins are transmembrane proteins mediating water transport across plasma membrane of animal, vegetal or bacterial cells. Among the ten aquaporins known in mammals, six are located in kidney and take part in urine concentration. AQP2 is vasopressin regulated, it is the only family member to be implicated in human pathology, such as nephrogenic diabetes insipidus, congestive heart failure, hepatic cirrhosis, nephrotic syndrome or SIADH. Aquaporins are expressed in a wide variety of tissues, such as brain or gastrointestinal tractus, and suggest a role in water tissue exchange, but their real function is still not define. To know the physiological impact of aquaporins, AQP1, AQP3, AQP4 and AQP5 knockout mice have been created and their phenotype analysed. PMID- 10667047 TI - [Surgery, radiotherapy or hormonal therapy in the treatment of prostate cancer]. AB - Prostatic cancer (PC) became the first diagnosed cancer in western men and is the second leading cause of cancer death in men. Wide utilisation of serum PSA and free PSA measurements, identifies patients requiring transrectalultrasonography (TRUS) and TRUS guided biopsies. Most prostatic cancers diagnosed today are locally limited and may be treated by radical surgery or radiotherapy. In case of disseminated disease, hormonal manipulations remain the treatment of choice. In that field, many new drugs have been designed to allow medical castration with less complications, especially regarding sexual potency. PMID- 10667048 TI - [Treatment of border-line dilatation of the ascending aorta using the Robicsek intervention]. AB - The technique and the results of the external contention of the dilated ascending aorta by the application of an external Dacron graft are reported. This technique, without replacing the usual graft interposition, can efficiently and expeditiously treat borderline dilatation of the ascending aorta, as frequently encountered in aortic valve disease. If the sinotubular junction is not dilated, this technique may stop the aneurysmal evolution of the ascending aorta. PMID- 10667049 TI - [Results of artificial sphincter for the treatment of sphincter incontinence]. AB - 47 patients were treated since 1987 by implantation of an artificial sphincter. The majority of female patients were operated for bladder neck hypermotility. In the male population, incontinence was mostly a complication of prostatic surgery. The indication, preoperatory assessment, surgical techniques and results are discussed. During a mean follow-up of 38 months, 92.9% of females remained perfectly continent and results were good in 76.7% of males. The results are similar to those reported in the literature. PMID- 10667050 TI - [How I assess ... home glycemic control in diabetic patients]. AB - Self blood glucose monitoring is now a key element in the management of diabetic patients. However, in order to improve overall blood glucose control, self monitoring should lead to self-management. This implies that the patient modifies his/her behaviour (diet, exercise, oral antidiabetic agents, insulin doses) to ameliorate the metabolic status according to his/her blood glucose measurements. While the place of home blood glucose monitoring is rather well defined in type 1 diabetes, it remains more controversial in type 2 diabetes. PMID- 10667051 TI - [Clinical study of the month. Does chronic sleep deprivation predispose to metabolic syndrome?]. AB - According to a recent experimental study published in the Lancet, sleep debt, frequently imposed by the life habits of industrialized countries, results in profound metabolic (impaired glucose tolerance) and endocrine (increased sympathetic activity and evening cortisol levels) alterations, which mimic those of normal ageing and may have pathophysiological consequences in the long term. Another study recently published in the International Journal of Obesity demonstrated a significant positive relationship between the duration of shift work and body mass index or waist to hip ratio, a marker of visceral adiposity. One may thus hypothesize that chronic sleep deprivation could predispose to the metabolic syndrome and increase the cardiovascular risk. PMID- 10667052 TI - [The incineration of waste: a poisonous debate?]. PMID- 10667053 TI - [Are sanitation services complaints an indicator of quality of care?]. AB - A retrospective analysis of 211 consecutive complaints treated at the Direction of Health and Social Assistance of Paris was undertaken in order to specify the nature of the complaints and to evaluate their pertinence as an indicator of quality of care. The majority of complaints concern public and private health establishments, in particular surgery and psychiatric services. Although the study confirms the dysfunctioning of the organisation of services and also of therapeutic methods and medical treatments, the evaluation of iatrogenic risks and their avoidable nature is difficult and requires precise instruction. Complaints seem to be a neglected indicator of quality, yet they concern information that is accessible and could, if used with other information, be a first milestone in the vigilance of medical treatments. PMID- 10667054 TI - [Situation and role of local structures of hospital ethics: s survey of Hospital Public Care in Paris]. AB - This article presents the main results of a survey carried out among the local structures for hospital ethics of the Public Assistance sector of the Hospitals of Paris. The results show that the situation of these structures has completely transformed itself since the law of 1988 on human research. Only four ethics committees subsist out of the 16 university-hospital committees that existed in 1991. Seven new structures have been created since 1994, which are open to all categories of personnel and within which doctors are a minority. The four ethics committees provide almost no decision making opinions anymore and are, like the new groups, fora for discussion, reflection, or even information on issues of hospital ethics. Almost none of these structures has an official mission. This situation presents the question of the role that a local ethics structure can hold within a hospital. PMID- 10667055 TI - [Feasibility of drug use detection among individuals 15-25 years of age in community medicine]. AB - The objective of this work is to study the feasibility of the detection of drug use or misused substances during a visit with a municipal doctor, and to describe the interrogatory strategies used spontaneously by doctors. One tenth of the clients aged 15-25 of the 26 participating doctors, (the majority of them men visiting the doctor for a psychological problem), showed a "warning" signal and were detected. The doctors often chose to approach the problem of drugs directly and detected usage in two-thirds of the cases. Cannabis was most often concerned, but the use of psychotropic drugs with alcohol was the case in one-third of the subjects interrogated. On the other hand, a certain frustration was felt by the doctors because of the difficulty of approaching the root problems and of acquiring psychotherapeutic monitoring. PMID- 10667056 TI - [Contributions to the sociologic analysis of the impact of sickle cell disease on families from Northern Tunisia]. AB - The improvement of the conditions of care and quality of life of people living with drepanocytosis now constitutes a major concern of health authorities and voluntary groups (associations of sick people and their parents). In order to examine the conditions and methods of care for children living with drepanocytosis in the Bizerte region of Northern Tunisia and to understand the problems and difficulties experienced by them and their families in their daily lives, an anthropological study was carried out in the region between January 16 and February 12, 1997. Thirty-three interviews were carried out with families of infected children. The interviews were semi-directive using a pre-established protocol (interview guide). Thirty interviews were then analysed. A multitude of information concerning various medical and psycho-social aspects of drepanocytosis in the region were revealed. The data were regrouped into the following categories: circumstances of discovering the illness; explicative model of the illness as perceived by the people interviewed; conditions and methods of care of the sick child; degree of satisfaction with the health care system; psychological profile of the children interviewed; educational level of the children interviewed; repercussions the illness has on the family; impact of the illness on household budget; suggestions and comments of the people interviewed. In light of the information collected, various actions have been proposed with the goal of improving the conditions of care and quality of life of people living with drepanocytosis in the region. PMID- 10667057 TI - [Occupational hazards in the Moroccan craft sector and proposal for occupational health services]. AB - The absence of occupational health services, the numerous occupational hazards and the high number of people working in the handicraft sector have sparked this study. Descriptive, cross-sectional epidemiological studies were carried out throughout 1996 for different artisan activities: iron-work, jewellery making, rug making, tannery, "zellige", pottery, and woodworking. The study included 449 artisans and consisted of an analysis of work conditions, a medico-social questionnaire, a clinical examination and among certain artisans, a biological check-up, a respiratory check-up (thoracic x-ray and lung function testing), and a toxicological check-up. Poor work conditions and the absence of any technical protection (collective or individual) are common to all the workshops visited. Multiple risks as well as various and frequent pathologies were observed for all the artisan activities. The most common ailments are those linked to posture and musculo-skeletal problems (67.6%), oral (58.2%), ocular (46.9%), dermatological (35.7%), ear/nose/throat (35.3%), respiratory (31.1%), digestive (21.1%) and neurological (20.7%). Often the same artisan showed several simultaneous conditions linked to work. The legislative texts related to occupational health and safety are many and dispersed, and would profit by being updated and regrouped within a work code which would make them easier to consult and would allow all partners in the social sector to get to know them. In addition, this legislation which has been strengthened, is unfortunately not enforced. We should support every initiative focused on developing the prevention of occupational hazards and the spirit of safety within artisan workshops. The concern for occupational risks within the artisan milieu owes its importance to their abundance, the diversity of the professions involved, and the number of different risks to which artisans are exposed. The role of a worksite doctor is therefore considerable, and his field of intervention in this milieu is vast. The broader goal of occupational health services is to protect and improve the physical, mental, and social well-being of its workers; it is natural that these services should give more attention to general health promotion (vaccinations, health education...). Given that the artisan sector is organised around its structures of production, it seems urgent to introduce medical coverage and to improve health and safety conditions within the sector. PMID- 10667058 TI - [Palliative care at a university hospital center: physicians' opinions (1992 1996)]. AB - OBJECTIVE: To evaluate the evolution of doctors' opinions in a university hospital centre concerning care for terminally ill patients following the creation of a Mobile Palliative Care Unit (MPCU). METHODS: In the relevant services of a university hospital in Paris (1095 beds), (1) the first survey in 1992 included interviews with 17 doctors; (2) the second interview was carried out in 1996 by self questionnaire among 55 doctors. RESULTS: (1) The doctors stressed their wishes to accompany their patients "until the end" and stressed the role of the patients' family (support system) and of nurses. They indicated their wishes especially for training then for the creation of a MPCU. (2) The MPCU, solicited by 87% of doctors, teaches palliative care and provides therapeutic, psychological and relational assistance, judged as very satisfactory, to carers and families. The doctors would like to see these activities sustained. PMID- 10667059 TI - [Physicians' opinions and practices on the use of medical referrals at hospitals]. AB - This work aims to take stock of the actual utilisation of medical standards (references), through a study of opinions and practices of the medical managers working in the clinical services of a university hospital centre. A survey through interviews was proposed to 103 medical managers, 101 responded to the questionnaire (38 Unit directors and 63 directors of "UF" units). Medical standards are used essentially as a teaching aid by 80% of doctors. Some of them (48%) make them available to prescribing doctors in the unit, and for 36% there exists an informational procedure for new residents. Evaluation studies concerning the implementation of medical standards remain rare (7 studies). Medical standards appear to be more useful for improving quality of care (90%) then for controlling health expenditures (72%). The majority of medical managers (72%) consider that certain standards should be opposable to hospitals. The medical managers of the university hospital centre are in favour of developing standards of clinical practice. PMID- 10667060 TI - [Effectiveness of a program for improving drug prescriptions and hospitalization reporting at a university hospital]. AB - OBJECTIVES: To evaluate the effectiveness of a method for improving the quality of care applied to the formulation of medical prescriptions and to the release of hospitalisation reports. The actions comprised the diffusion of results of audits and recommendations of good practice. METHODS: For each of the 41 services, 30 visits from 1996 were selected at random. 3289 prescriptions and 1067 correspondence files were analyzed. The results were compared to those obtained from the previous two years. RESULTS: Patient identification was complete in 44% of prescriptions, the identification of the prescribing doctor and his signature were present in 62% and 19% of cases respectively. 37% of medicines included all information. 7 indicators out of 12 for the quality of prescriptions improved (p < 0.0001). Files were found for 83% of hospital visits and 56% were sent (released) within a week. The practitioner was identified in 79% of cases, the main diagnosis in 96% and the treatment in 65% of cases. Five out of nine indicators of the quality of correspondence improved (p < 0.01). For each theme, the number of indicators improving was similar (p > 0.05). The services that improved for one theme didn't necessarily improve for the second (p > 0.05). DISCUSSION: The evaluation of the programme, based on a strategy of quality assurance, shows modest progress. Given that the improvement of two themes for a given service are not correlated, the programme appears to sensitise professionals at an individual level rather than collectively. However, this programme is an important step for introducing a mode of continued improvement of quality. PMID- 10667062 TI - [The internet and public health: access to information]. PMID- 10667061 TI - [Lack of communications between general medicine and the mental health sector]. AB - Mental health problems today account for a rising number of visits to general practitioners, which require collaboration between general practitioners and psychiatrists. A KAP study (Knowledge, Attitudes, Practice) was carried out among general practitioners of three towns in the "Meurthe et Moselle" region, in the common territory shared by both a child and an adult psychiatric sector. Twenty three doctors participated in the study. General practitioners know the different psychiatric structures but they do not know their areas of speciality or how they are organised. All recognise the high frequency of mental health problems among their patients, the most frequent being depressive pathologies. They have a negative image of the sector which is perceived as a complex "world" from which they feel excluded as soon as they refer a patient because of the difficulty they have in communicating with psychiatrists. PMID- 10667063 TI - [When ideology is a barrier to violence prevention]. PMID- 10667064 TI - [Introduction to molecular and cellular biology of chloroplast biogenesis]. PMID- 10667065 TI - [Structure and function of the plastid-dividing apparatus]. PMID- 10667066 TI - [Molecular organization and dynamism of chloroplast nucleoid, a eukaryo prokaryotic hybrid]. PMID- 10667067 TI - [Transcriptional regulation in the chloroplast: mechanisms of coordinated expression of photosynthesis genes]. PMID- 10667068 TI - [Post-transcriptional regulation of chloroplast gene expression]. PMID- 10667069 TI - [Protein transport of chloroplast proteins]. PMID- 10667070 TI - [Chloroplast function and plant development]. PMID- 10667071 TI - [Progressing molecular cell biology in bacteria: chromosome partitioning and cell division]. PMID- 10667072 TI - [The role of paraxial protocadherin in morphogenetic cell movement during gastrulation]. PMID- 10667073 TI - [Imaging and manipulation of DNA molecules using scanning tunneling microscopy]. PMID- 10667074 TI - [Deblocking aminopeptidase from Pyrococcus furiosus and its application for proteomics]. PMID- 10667075 TI - [Check-up and screening]. PMID- 10667076 TI - [Check-up examinations from the cardiologic viewpoint]. AB - The concept of the primary prevention of coronary disease is certainly widely accepted today and supports check-up visits of apparently healthy persons. The goals of check-up's viewed by the cardiologist are the detection of coronary risk factors and the identification of asymptomatic patients with coronary disease to initial preventive measures. In some instances, a personal health problem forces the patient to a check-up visit. The major coronary risk factors of heredity, cigarette-smoking, hypercholesteremia, hypertension can be detected by taking the history, performing a physical examination and a blood-sampling for cholesterol. Additional investigations such as an ECG or an exercise-test are only indicated in symptomatic patients or in persons at high coronary risk. PMID- 10667077 TI - [Screening, diagnosis and management of diabetes mellitus and diabetic complications]. AB - Diabetes mellitus comprises a group of metabolic disturbances that are characterized by hyperglycemia. In 1997 the American Diabetes Association (ADA) proposed new criteria for the diagnosis and classification of diabetes mellitus, which was also adopted by WHO. Although the criteria is the same, the ADA puts emphasis on the use of the fasting plasma glucose (FPG) for screening and diagnosis, whereas WHO maintains the use of the OGTT and recommends the FPG only if an OGTT can not be performed. Different pathogenetic processes are involved in the development of diabetes ranging from autoimmune destruction of beta-cells resulting in an absolute insulin deficiency to insulin with a defect on insulin secretion. The new classification is based on the etiology of the disease. Diabetes is classified into one of four categories: Type-1, type-2 Diabetes mellitus, specific forms of diabetes, and gestational diabetes. For screening and diagnosis FPG or the two hour value after the OGTT can be used. Glycosylated hemoglobin is not suitable for screening and diagnosis of diabetes despite some contradictory statements. For many decades clear evidence was missing that chronic hyperglycemia caused diabetic late complications; complications including dysfunction or failure of several organ systems, in particular eyes, kidneys, nerves, and the cardiovascular system. The results of two large prospective trials--the Diabetes Control and Complications Trial (DCCT; 1993) and the United Kingdom Prospective Study (UKPDS; 1998)--that were recently published provided the final proof that normoglycemia prevents or delays the progression of these late complications. Due to the insidious nature of these complications they are often not diagnosed and have to be looked for in each patients with diabetes and have to be controlled regularly. Based on the results of the UKPDS and other studies, evidence based therapeutic goals could be defined. Multifactorial interventions with increased physical activity, cessation of smoking, aspirin treatment, lowering of HbA1c, blood pressure, and lipids in patients with type 2 diabetes have been proven to drastically reduce the risk of developing diabetic nephropathy or cardiovascular complications drastically. We recommend the following treatment strategy for patients with type 2 diabetes in clinical practice: 1) Treatment should be individualized. 2) Treatment should be started step by step to document efficacy of treatment and compliance of patients. 3) Plasma glucose and blood pressure should be normalized in all patients with type 2 diabetes (up to an age of 70 years), since there are no threshold values for HbA1c and blood pressure. 4) Therapeutic goals should be checked every three to six months. 5) In the case that therapeutic goals can not be met, treatment should be intensified. Often a combination therapy with many different drugs is required. 6) A specialist for diabetes should be consulted, if the therapeutic goals can not be met over a period of six months. PMID- 10667078 TI - [Skin check-up--who and when?]. AB - The skin is easy to access and therefore this organ is also an important diagnostic tool for the general practitioner to establish a systemic and complex disease. In addition skin cancer is the most prevalent malignancy and therefore non-dermatologists should be able to make the diagnosis of an early skin cancer. It is recommended to perform a whole body examination in all first visits. With experience a whole skin check lasts no longer than five minutes. It is important to recognise patients at risk for skin malignancies because they clearly profit from a regular screening. Patients with atypical moles or with familial melanoma syndrome and patients who already had malignant melanoma should get a yearly skin examination performed by a trained expert. Patients who had basal cell carcinoma run an almost 50% risk to produce a second basal cell carcinoma within three to five years and therefore they are good candidates for a screening at a regular base. The number of organ transplanted people is rising dramatically. In general these patients are under marked immunosuppression and therefore they often develop malignancies of the skin. In our Department such patients receive at least one dermatological visit a year. PMID- 10667079 TI - [Screening for prevention and early detection of colorectal carcinoma]. AB - Colorectal Cancer (CRC) fulfills all the criteria for a successful screening: i) it is a frequent tumor associated with serious morbidity and mortality; ii) recognition and treatment of premalignant lesions or early tumor stages improves prognosis and lowers its incidence, respectively. During recent years, our understanding of the biology, diagnosis and therapy of adenomas and CRCs led to the identification of risk factors. Based on these, screening strategies differ for individuals with average risk (age > or = 50 years, no risk factors) and individuals with increased risk for CRC development (positive personal history [CRC, colorectal adenomas, inflammatory bowel disease]; positive family history [CRC, colorectal adenomas, hereditary syndromes]). The currently available strategies aimed at prevention and early detection of CRCs (past history, family history, rectal digital examination, fecal occult blood testing, endoscopy) are discussed and recommendations for their implementation are formulated. PMID- 10667080 TI - [Prostatic carcinoma. Screening--when and what?]. AB - Prostate cancer is now the most common cancer and the second most common cause of death from cancer among men. Therapy of curative intention is only possible in organ confined disease. The use of prostate specific antigen (PSA) and digital rectal examination (DRE) results in a three fold increase in prostatic carcinoma detection. Levels of PSA > 4 ng/ml are indications for sextant biopsies of the prostate. There did not exist an intermediate range or 'grey zone' of PSA 4-10 ng/ml where wait and see diagnostic procedure is indicated. In PSA levels > 10 ng/ml curative therapy can only performed in 15-44% of the cases. PSA and DRE examination should be performed between the age of 50 and 70 years when life expectancy exceeds ten years. In case of familiar history the case finding has to start at the age of 45. There is no support for the common opinion that early detection finds clinically insignificant cancer since autoptical prevalence of prostate cancer is about 40% and early detection discover only 3-4%. Results about the usefulness of active screening in a population will be available in 2005. PMID- 10667081 TI - [Screening for gynecologic-endocrinologic problems before menopause]. AB - Family and personal history as well as clinical examination are the basic data to be known before laboratory examinations should be started. To obtain results that can be correctly interpreted, the blood sampling has to be done in the early morning hours between day 1 and 5 of the cycle, and for some hormones on an empty stomach. Depending on the clinical data, the hormonal screening can be selective and well directed, or it has to be broader. The presence or absence of galactorrhea, of hot flushes and of androgenization or virilization play an important role for the decision about the hormones to be determined. Furthermore, an eventual desire infertility will influence the selection of the hormonal tests to be done. The present review intends to propose some simple recommendations to the non-specialist how a gynaecological-endocrinological screening for the most important clinical questions should be organized. PMID- 10667082 TI - [Guidelines for good practice: anti-ulcer agents, indications in the adult. French Agency for Health Safety of Health Products]. PMID- 10667083 TI - [Outline of the problem of indices of therapeutic efficacy. 4. Expression of efficacy when the underlying illness is incurable. Study Group for the Indices of Efficacy]. AB - In chronic illness, when death or a non-fatal event can occur at any time, the current efficacy indices are no longer appropriate to express the effect of the treatment on the potential therapeutic objectives. The inappropriateness is not dependent on the effect model. Clues for solutions are proposed. PMID- 10667084 TI - Adverse drug reaction monitoring and the Internet: evaluation of the use of the Internet by French Pharmacovigilance Centres and a non-exhaustive survey of websites of interest for collecting information about adverse drug reaction. AB - The Internet, indisputably the most important source of information obtainable in real time, was long essentially the domain of researchers in the USA, but has now become more accessible to French Pharmacovigilance Centres (CRPV) and pharmaceutical companies. A questionnaire was sent to every CRPV to determine how the Internet was perceived in terms of pharmacovigilance activities and existing Websites of particular value to CRPVs were investigated. Analysis of the questionnaires revealed that 66.7 per cent of CRPVs are connected to the Internet but do not use it fully; 94 per cent of connected CRPVs use it essentially for access to a bibliographic database such as Medline (88 per cent), and none subscribes to discussion lists concerning alert messages on problems related to drugs and therapeutics. Non-connected CRPVs do not intend to use the Internet because of financial considerations, lack of time or the assumption that it is not beneficial in everyday situations. Apparently, many French CRPVs are not sufficiently aware of the importance of the Internet for professional purposes. A non-exhaustive list of sites on the Internet providing information likely to be of use to Pharmacovigilance Centres in their everyday activities is included. The Internet offers far greater possibilities than research for bibliographic references on Medline and could improve the manner in which pharmacovigilance is practised. PMID- 10667085 TI - [H1 antihistamines: the past and the future]. PMID- 10667086 TI - [Tomorrow's drug surveillance]. PMID- 10667087 TI - [Iodine status and the used of iodized antiseptics in the mother-newborn pair]. AB - Iodine status was evaluated by assessment of urinary iodine excretion in 221 mothers and their 223 newborns. During the first month after childbirth, 59.3 per cent of the mothers and 26.5 per cent of the newborns received applications of iodized antiseptic containing Polyvidone-iodine. 50.2 per cent of the newborns and 24.9 per cent of the mothers had a urinary iodine of more than 20 micrograms/dl (iodine excess). For the mothers and the newborns who had received applications of iodized antiseptic, 38.2 per cent and 74.6 per cent had an iodine excess, respectively. This iodine excess is directly related to use of iodized antiseptic. Such high iodine levels may contribute to the risk of thyroid disorders, and particularly to transient congenital hypothyroidism at a critical age for normal development of the nervous system. PMID- 10667088 TI - [Use of iodized salt and the risk of iodine overload]. AB - Iodine-deficiency disorders are a major problem of public health in Morocco. To mitigate this deficiency, the iodination of all the salt intended for human consumption in a proportion of 80 +/- 10 mg/kg of salt has become obligatory since a decree published in 1995. We estimated that this rate of iodized salt issued risked inducing an iodine excess in the population. To check this hypothesis, we provided 7 families made up of 28 subjects, who at the start were consuming a non-iodized salt, with the decreed, iodized salt and we followed the evolution of their urinary iodine excretion over a period of 3 weeks. The mean values of urinary iodine excretion of the 28 subjects were 12.8 micrograms/dl before use of iodized salt and 26.8, 35.5 and 63.2 micrograms/dl, respectively, after 7, 14 and 21 days from the introduction of iodized salt into their diet. After 21 days of the use of iodized salt, 84.6 per cent of the subjects had an iodine excess. We conclude that prolonged use of this iodized salt exposes the population to the risk of thyroid disorders. PMID- 10667089 TI - Effect of dietary crude proteins on the reproductive function in the postpartum dairy cow. AB - The study was conducted on 216 dairy cows. Samples of feeds distributed to cows were collected monthly for the purpose of determining their content in dry matter, energy, crude proteins and mineral matter. Milk samples were collected weekly for every cow from newly calved cows until confirmation of pregnancy by rectal palpation at least 2 months after artificial insemination. These samples were used for progesterone assays in skimmed milk, in order to assess the interval between calving and return to ovarian activity [C-ROA], calving and first insemination [C-I1], calving and conception [C-C] and number of inseminations per conception (nI/C). Results have shown a significant negative correlation between the duration of [C-ROA] and [C-C] intervals and the dietary content in crude proteins (r = -0.720, p < 0.05 and r = -0.914, p < 0.01 respectively). PMID- 10667090 TI - New model of atherosclerosis in sand rats subjected to a high cholesterol diet and vitamin D2. AB - In order to defeat the atheroresistance in sand rats, 25 animals were given a high cholesterol diet for 45 days, which was then associated with oral treatment with vitamin D2 2000 IU/day for a further 45 days. At days 0, 45 and 90, plasma parameters, and aortic and heart morphology were examined. Results showed at D45 hypercholesterolaemia, increased plasma LDL and VLDL cholesterol, oxidized LDL, triglycerides, free fatty acids (FFA) and calcium levels and moderate hyperinsulinaemia. At D90, plasma-oxidized LDL and FFA were more enhanced, whereas calcium level was reduced. Development of hyperglycaemia was associated with hyperinsulinaemia and insulin resistance. The vitamin D2 administration induced advanced lesions, represented by the degenerescence of elastic lamina, smooth muscle cell proliferation and lipid calcic plaque at an ulcerated stage in most cases. The ischaemic effects were represented by acute myocardial infarction. The potential of the sand rat to develop atherosclerotic lesions at different stages opens the field to therapeutic tests of new anti-atherogenic agents. PMID- 10667091 TI - Epidemiological study: chronotype and daytime sleepiness before and during Ramadan. AB - Few epidemiological data have been reported on the relation between Ramadan fasting, life habits (meal frequency, sleep habits) and daytime sleepiness during Ramadan. This paper presents the results of a detailed study of the chronotype and daytime sleepiness before and during Ramadan. It was conducted on a sample of 264 subjects aged between 20 and 30 years. Results have revealed a significant decrease in the meal frequency during Ramadan compared with the control period. Before Ramadan, the majority of subjects woke up between 6 and 7 a.m. and went to sleep between 10 and 11 p.m. however, during Ramadan fasting, they woke up after 8 a.m. and preferred to go to sleep later (after midnight). Chronotype as evaluated by the Horne and Ostberg scale was changed significantly during Ramadan: an increase of the evening type and a decrease in the morning type of subjects was observed. Daytime sleepiness as evaluated by the Epworth Sleepiness Scale was significantly increased. PMID- 10667092 TI - Calcitonin-induced impairment in conditioning is antagonized by chronic antidepressant drug treatment. AB - The purpose of this study was to test the effect of calcitonin, when injected into the lateral ventricle, on conditioning behaviour and to see whether antidepressant drug treatment can antagonize calcitonin-induced impairment of this behaviour. Conditioned response by conditional stimulus (CS) was compared in control rat (CO) and in rats that received intraventricular perfusion of calcitonin (CA), acute antidepressant drug treatment (ADa), acute antidepressant drug treatment + calcitonin (ADa + CA), chronic antidepressant drug treatment (21 days) + calcitonin the day after (ADc + CA). Control rats acquired easily the conditioned response, the CA group and ADa + CA had problems in making the correlation between CS and unconditional stimulus (US), and consequently did not acquire a conditioned response, but in the ADc + CA group, rats exhibited more conditioned responses. The results indicate that calcitonin disrupts conditioning processes and chronic but not acute antidepressant drug treatment can reverse the effects of calcitonin. PMID- 10667093 TI - Modulation of central GABAA receptor complex by somatostatin: a pharmacological study. AB - The goal of the present study was to determine the possible interactions between somatostatin (SST) and gamma-aminobutyric acid (GABA). We thus investigated the SST interaction with [35S]-tertiary butylbicyclophosphorothionate (TBPS) binding sites of the cortical and hippocampal regions of the rat brain. The method used to identify such effects is in vitro quantitative autoradiography. Thus, the binding of the cage convulsant [35S]-TBPS to a picrotoxin-sensitive site in the rat brain was used to investigate the modulatory action of SST on the GABAA receptor complex. The addition of the peptide to the incubation medium results in a dose-dependent inhibition of [35S]-TBPS in cortical and hippocampal structures. Detailed analysis showed a dose-related effect of SST with relative potencies comparable to those observed for 5 alpha 3 alpha P and 5 beta 3 alpha P. In addition, these neurosteroids were able to enhance the efficacy of SST in inhibiting [35S]-TBPS binding. The efficacy of SST in enhancing the inhibitory action of neurosteroids was also evidenced. Furthermore, SST seems to mimic the effects of these neurosteroids as well as GABA and picrotoxin on [35S]-TBPS binding to the rat brain in every context examined. This suggests that somatostatin allosterically modifies [35S]-TBPS binding through a mechanism similar to that of GABA. On the other hand, a possible action of SST via transduction systems on the GABAA receptor complex could also be suggested. These results illustrate the importance of interactions in SST-mediated GABA transmission in these brain regions. PMID- 10667094 TI - Interaction of chlorpromazine and imipramine with model membranes. AB - The aim of the present study is to examine the effect of phospholipids on the lipid destabilization induced by chlorpromazine (CPZ) and imipramine (IP) at different levels of pH. The large unilamellar vesicles (LUV) are formed in the presence of calcein (60 mM). The vesicles containing calcein have been incubated in the presence of CPZ and IP at pH 4.5 and pH 8. At pH 4.5 CPZ and IP induce a rapid release of the calcein encapsulated in the liposomes. Calcein release, at equal concentrations of pharmacological agent, is more important by CPZ than by IP. At pH 8, the calcein release was more important than at pH 4.5; this effect appears to be more significant for the CPZ than for the IP. In conclusion, the insertion of chlorpromazine and imipramine into large unilamellar vesicles is accompanied by a strong destabilization of the vesicles. These effects appear more significant for chlorpromazine than for imipramine. PMID- 10667095 TI - Myoclonic and tonic seizures elicited by microinjection of cholinergic drugs into the inferior colliculus. AB - The inferior colliculus (IC) is the initiation site in the neuronal network for the epileptic audiogenic seizure (AGS). The present study investigates the effects of alteration of IC cholinergic transmission on the elicitation of epileptic seizures. Unilateral microinjections of carbachol (3 and 6 micrograms/0.2 microliter) into the IC elicited intense locomotor activity, contraversive rotations and myoclonic seizures. This result indicates that the IC is the initiation site for the induction of myoclonic seizures and suggests that these myoclonic seizures may result from activation of m1 muscarinic receptors. Microinjections of the nicotinic-muscarinic antagonist, gallamine (2 and 6 micrograms/0.2 microliter), into the IC induced AGS susceptibility. However, microinjections of muscarinic antagonists, atropine (15 micrograms/0.2 microliter) and scopolamine (12 and 20 micrograms/0.2 microliter), or the nicotinic antagonist, hexamethonium (12 and 20 micrograms/0.2 microliter), into the IC have no effect. Gallamine-induced AGS susceptibility may result from a selective blockade of m2 muscarinic receptors. PMID- 10667096 TI - Interactions between neomycin and cerebral dopaminergic and serotoninergic transmission in rats. AB - The aim of this work was to investigate the effects of neomycin on monoamine contents of rat striatal tissue. The analysis was performed by HPLC with electrochemical detection. Rats were injected with neomycin (2 mg/kg; i.p.) and killed at different times over 24 h. The striatal homogenate was injected in a reversed-phase HPLC. Results showed a significant increase in dopamine tissue level (+35 per cent; 4 h after neomycin injection) and a decrease (-36 per cent) in the level of its metabolites. Striatal serotonin level showed a rapid and significant (p < 0.001) increase, +130 per cent, 2 h after neomycin injection. DOPAC/DA and 5HIAA/5HT ratios were reduced by 34 per cent and 46 per cent respectively. These results indicate an inhibitory effect of neomycin on striatal dopaminergic and serotoninergic systems. Several mechanisms could be involved in these effects of neomycin in the biosynthesis process through stimulation of tyrosine hydroxylase and tryptophane hydroxylase and/or MAO and COMT activities. The blockage of calcium channels was also suggested. PMID- 10667097 TI - Comparative effects of UW and SLS solutions on concentrative proline uptake in cold preserved rat hepatocytes. AB - In previous studies, we have shown that the rate of cell swelling induced by concentrative proline uptake in isolated rat hepatocytes decreased by 50 per cent after only 24 h of cold storage in University of Wisconsin (UW) solution, thereby representing a sensitive marker of alterations in hepatocyte functions after cold preservation and rewarming. We have thus used concentrative proline uptake to compare the capacity of UW and sodium-lactobionate-sucrose (SLS) solutions to maintain such differentiated hepatocyte functions. Isolated rat hepatocytes were kept at 4 degrees C for 4, 10, 24 and 48 h in UW or SLS solutions, and subsequently cultured at 37 degrees C for 1-2 h. Viability was measured by Trypan blue exclusion. After rewarming, cells were subjected to a 10 min administration of 10 mM proline and accumulation of the amino acid was assessed by changes in cell volume as measured by digital analysis of single-cell images obtained under bright-field illumination. Cell viability was reduced gradually and significantly after 0 to 48 h of preservation, and rewarming amplified this effect. However, loss of viability was similar in UW- and SLS-stored cells, as were initial steady state cell volumes. Proline-induced swelling rate was reduced significantly by 13, 46 and by 57 per cent after 10, 24 and 48 h of preservation in UW solution, respectively. There is no significant difference between SLS- and UW-preserved hepatocyte swelling rates after 10 h and 48 h of cold preservation. However, the decline in the swelling rate of SLS-preserved hepatocytes incubated for 24 h is significantly lower than that of their UW-preserved counterparts. These results show that the SLS solution can preserve differentiated hepatic functions as well as the UW solution does. PMID- 10667098 TI - In vitro effects of spiramycin and dirithromycin on IL1 beta production by human LPS-stimulated mononuclear cells. AB - Polymorphonuclear neutrophils are the predominant cells in acute inflammatory lesions and their functions and recruitment are regulated by cytokines, including IL1, TNF and IL8. Antibiotic modulation of inflammatory effects has stimulated investigations of antibiotics for their potential activity as immunomodulators over their primary bactericidal or bacteriostatic activities. This study reports the influence of macrolides, spiramycin and dirithromycin on IL1 beta production. Mononuclear cells, isolated from healthy human volunteers, were preincubated with macrolides (0.1 to 500 micrograms/ml) and stimulated by Escherichia coli lipopolysaccharide. Then, IL1 beta production was detected by western blotting analysis. At therapeutic concentrations, dirithromycin and spiramycin seemed to enhance IL1 beta production by LPS-stimulated cells, with +37 per cent and +28 per cent at 1 microgram/ml respectively. At supratherapeutic concentrations, these drugs seemed to inhibit IL1 beta production through protein kinase C inhibition, with inhibitory concentrations 50 per cent of 378 micrograms/ml for dirithromycin and 234 micrograms/ml for spiramycin. So, macrolides may modulate the host defence system through their influence on cytokine production. PMID- 10667100 TI - Mycobacterium resistance to antimycobacterial reagent. AB - This study was carried out in the tuberculosis laboratory of the Institut National d'Hygiene in Rabat, Morocco, in 1997. The aim was to determine the percentages of drug-resistant strains by using 150 antibiograms. Six antimycobacterial medications were used as tuberculosis treatment: isoniazid (INH), streptomycin (STM), rifampicin (RIF), ethambutol (EMB), kanamycin (KAN) and p-amino-salicylic acid (PAS). The cultures were plated onto a simple agar (Lowenstein-Jensen) plate containing different concentrations of drugs. This test demonstrated the presence of major antimycobacterial (INH, RIF, STM)-resistant strains of Mycobacterium tuberculosis in the following percentages respectively: 34.6 per cent, 33.1 per cent and 26.1 per cent and 80 per cent, 70 per cent and 40 per cent in the case of atypical mycobacteria. The association of INH/RIF showed the highest percentage (27.6 per cent) for Mycobacterium tuberculosis and 70 per cent for atypical strains, whereas, when we associate INH/RIF/STM, the resistance rate becomes 17.69 per cent for Mycobacterium tuberculosis and 25 per cent for atypical mycobacteria. The resistance in question was a secondary or acquired resistance because the tested strains were isolated from patients who had not responded to standard tuberculosis treatment. PMID- 10667099 TI - A comparison study of two therapeutic protocols for neurosyphilis. AB - In Morocco, neurosyphilis is a serious public health problem. In the neurology service at the specialist hospital in Rabat, two drug treatments were used. Treatment A consisted of infusion over a period of 4 h of 20 MUI of penicillin G per day for 3 weeks. Treatment B consisted of infusion over a period of 6 h of 30 MUI of penicillin G per day for 10 days. Each treatment was tested on a group of eight neurosyphilitic patients as first-line treatment. On the first day of treatment, both blood and CSF pharmacokinetics were sampled for each patient. Blood and CSF were taken within 24 h. Penicillin G concentrations were determined by a microbiological method. The results obtained showed that perfusions of either 20 MUI or 30 MUI of penicillin allowed the achievement of high serum concentrations. These increased progressively until reaching their maximum at T4 h for treatment A (from 92.33 to 106.38 micrograms/ml). For treatment B, maximal concentration is obtained at T6 h (from 108 to 141.52 micrograms/ml). Penicillin concentrations decreased immediately after stopping the perfusion. At CSF levels, penicillin G concentrations were identical to serum concentrations. However, one difference was observed: a one-hour difference between the serum and CSF peaks. The CSF peak was achieved at T5 h for treatment A (0.063 to 2.25 micrograms/ml) and at T7 h for treatment B (0.92 to 2.94 micrograms/ml). The concentrations obtained were largely superior to the CMI of Treponema pallidum for both treatment A and treatment B, at 47 times and 82 times higher respectively. The recovery time of the patients was 14 h for treatment A and 24 h for B treatment. These results have shown that therapeutic method B was more efficient than A. Moreover, the evolution of penicillin G's diffusion in the CSF during treatment, of cell counts of protein level, of the VDRL test and of the gamma-globulin rate was studied. PMID- 10667101 TI - [Medicines interacting with mitochondria: anti-ischemic effects of trimetazidine]. AB - While mitochondria are key factors in energy production in cells they are also key factors in their life cycle because under certain circumstances they can provoke cellular apoptosis. Some 45 per cent of myocardial volume is taken up by mitochondria. Furthermore, mitochondria are key to many aspects of neuronal activity and can trigger neurodegenerative processes. Lipid oxidation is responsible for the production of much ATP resynthesis in the heart but this process is less oxygen efficient than glucose oxidation. During ischaemia, lipid oxidation is suddenly blocked, but markedly increased during reperfusion, causing accumulation of potentially toxic metabolites (acylcarnitines, acyl-CoA, lysophospholipids). These metabolites can change calcium handling, inducing arrhythmias. Trimetazidine, and another product in development, ranolazine, by inhibiting lipid oxidation favours glucose oxidation and inhibits the production of deleterious lipid metabolites. Thus this class of drugs can have beneficial effects on myocardial metabolism without direct haemodynamic effects. PMID- 10667102 TI - [Synthesis of benzimidazole derivatives by amino acid action on ortho phenylenediamine, variously substituted in positions 4 and 5]. AB - As part of the synthesis of the benzimidazole derivative heterocyclique system, we are interested in studying the condensation of the o-phenylenediamines with amino-acids such as aspartic acid, serine and histidine. The interest that these present is based mainly on their pharmacological properties. They have, in fact anti-inflammatory, antidepressive, antibacterial and antihistamine properties. On the other hand, it should be noted that 5,6-dimethyl-1-(alpha-D-ribofuranosyl) benzimidazole constitutes part of vitamin B12. Taken together, these results led us to pursue our research in this domain while focusing on new methods of benzimidazolic derivative synthesis. It should be said that the obtaining of these compounds depends on the quantity of the amino-acid. All synthesized products have been characterized by infrared, nuclear magnetic resonance and mass spectrometry. PMID- 10667103 TI - [Synthesis and pharmacologic study of 1,5-dialkyl-1,5-benzodiazepine-2,4 dithiones]. AB - The synthesis and psychotropic activity of 1,5-diakyl-1,5-benzodiazepine-2,4 dithiones (alkyl = methyl, ethyl and benzyl radicals) were studied. Alkylation reactions were performed in catalytic conditions by phase transfer. These reactions allowed us to isolate only one kind of product N-alkyl. Acute toxicity studies were conducted according to European protocols in two species of appropriate mammals in order to discover the lethal doses. The activity of the compounds on the CNS was then studied, using a battery of compartmental tests used in psychopharmacology. No toxicity was demonstrated at therapeutic doses. Each product had a sedative effect more or less pronounced and different from the reference substance clobazam (Urbanyl). They also had myorelaxant and anxiolytic effects, even lengthening the hypnotic effect of thiopental (synergic action). PMID- 10667104 TI - Isomers of substituted 3-benzo[b]furyl and 3-thienylaminobutyric acids as potent ligands of the GABA-B receptor: synthesis and preparative liquid chromatographic separation. AB - Baclofen (4-amino-3-(4-chlorophenyl)butyric acid) is the only selective agonist for GABA-B receptors. Its R-(-)-enantiomer is about 100 times more active than the S-(+)-enantiomer. In the search for new compounds that bind to GABA-B receptors, it is very important to clarify the structural requirements. The authors report the synthesis and separation of isomers of various 3 heteroaromatic (benzo[b]furan and thiophen) aminobutyric acids. The 4-amino-3-(7 methylbenzo[b]furan-2-yl)butanoic acid is a potent and specific ligand for GABA-B receptors, with an IC50 value of 5.4 microM for the displacement of [3H] GABA. PMID- 10667105 TI - Compatibility study of 5-fluorouracil with PVC bags after repackaging into two types of infusion admixtures. AB - The stability and compatibility of 5-fluorouracil (5-FU) in admixtures for continuous intravenous infusion using PVC bags and administration sets were studied. 5-fluorouracil was reconstituted and diluted to 1.8 mg/ml for infusion in polyvinyl chloride and glass containers, and to 1 mg/ml to 16 mg/ml for storage in polyvinyl chloride bags containing 5 per cent dextrose or 0.9 per cent sodium chloride injections. Admixtures were stored at +4 degrees C and with protection from light, for 7 days. Analyses were performed by HPLC. No significant drug loss was observed during simulated infusions using PVC infusion bags and administration sets vs glass bottles and administration sets, over an infusion period used in hospitals (24 h). The solution stored at +4 degrees C with protection from light in PVC bags showed that at 1 mg/ml to 16 mg/ml, 5 fluorouracil was stable at least for 7 days in 0.9 per cent sodium chloride and 5 per cent dextrose. PMID- 10667106 TI - [The problem of therapeutic efficacy indices. 3. Comparison of the indices and their use]. AB - Efficacy indices do not contain the same information although they are all combinations of the same two quantities. Therefore, one should choose the proper index. Actually, none is entirely appropriate. Each more or less meets the specifications, depending on the underlying effect model for the therapy considered. However, one can say that the absolute benefit is more appropriate from the patient's point of view, the relative from the scientific point of view and the number of patients to treat from the policy maker's point of view. Nevertheless, this classification needs to be considered with caution. Finally, it emerges from the review that none is fully relevant to express the efficacy of a therapy, even in the most suitable condition, the acute illness. PMID- 10667107 TI - [Constitutional thrombophilias: indications of the biological profile and therapeutic consequences]. AB - In laboratory screening in patients with clinical thrombophilia (early thromboembolism episode < 50 years, spontaneous thrombosis, recurrent thrombosis, unusual site of thrombosis, thrombotic family history or coumarin-induced skin necrosis complication), an isolated or combined inherited thrombophilia can be observed: antithrombin (0.5 to 4.9 per cent), protein C (1.4 to 8.6 per cent) and protein S (1.4 to 7.5 per cent) deficiencies or factor V Leiden (20 to 30 per cent). Special attention is mandatory in prescribing biological exploration because of the many physiological or pharmacological interferences which can modify the results. Identification of a genetic defect may induce specific management and individuals should receive counselling regarding the implications of this diagnosis. Further prospective studies should help to determine the thrombotic risk in symptomatic and non-symptomatic patients with inherited thrombophilia and the risk/benefit ratio of laboratory screening for hereditary thrombophilia and therapeutic intervention. PMID- 10667108 TI - [The rational basis for the short-, medium-, and long-term evolution of the pharmaceutical industry]. AB - In the last few years, a worldwide reorganization affecting the entire pharmaceutical industry has begun and is now accelerating. This intense trend of mergers, acquisitions, market entires++ and exits, reorientation and diversification reflects a new environment. This article sets out to explain the reasons behind this inescapable and rapid evolution. PMID- 10667109 TI - [How to improve drug development and utilization in pediatrics]. AB - Insufficient drug evaluation in children and the lack of adapted pharmaceutical formulations explain the importance of unlicensed and off-label prescriptions. As a consequence a regulation proposed in the USA by the Food and Drug Administration, requiring manufactures to assess the safety and effectiveness of new drugs in paediatric patients, has recently been adopted. Appropriate means to facilitate drug evaluation in children are now necessary in terms of recruitment and methodology. A ten-centre American Pediatric Pharmacology Research Unit network has been created and is being financed by the National Institute of Health. A similar trend is evolving in Europe. Appropriate drug utilization in children requires adequate formulations, administration devices and information as well as improved knowledge on the long-term potential consequences of drug use during growth and maturation. PMID- 10667110 TI - [Selective cyclooxygenase-2 (COX-2) inhibitors: importance and limitations]. AB - The discovery of an inducible form of cyclooxygenase (COX-2) requires a refinement of the theory that inhibition of cyclooxygenase activity explains both therapeutic effects and side-effects of non-steroidal anti-inflammatory drugs (NSAIDs). Selective COX-2 inhibitors have demonstrated in clinical trials a significantly better gastrointestinal tolerability than classical NSAIDs, for the same anti-inflammatory activity. Their tolerability in patients with active ulcer or with a recent history of ulcer as well as in patients suffering from cardiovascular or renal diseases has still to be investigated in detail. Their therapeutic potential in several new indications, including pre-term labour, colorectal cancer and Alzheimer's disease, is currently being investigated. PMID- 10667111 TI - [Research on inflammatory mediators in asthma: new therapeutic approaches]. AB - During the National Meeting of Clinical Pharmacology (Giens 1998), the working group exchanged information and examined the new therapeutic approaches to inflammation in asthma. The group performed a state of the art review of the current research on mediators of bronchial inflammation to determine new pharmacological targets of therapeutic interest. An update on the development of potential drugs acting on these targets was established. Specificities related to asthma in terms of evaluation and development of anti-inflammatory drugs were discussed. The place of pharmaco-economy in the development of anti-asthmatic drugs was specified. PMID- 10667112 TI - [The place of a new drug in the therapeutic strategy]. AB - A therapeutic strategy is a hierarchical set of appropriate measures to provide an answer to a pathological state. A drug is a part of this set (together with the diagnosis, the environment and the other medicinal interventions or not). A new drug's place in a therapeutic strategy can be evaluated according to one or several referential(s) when it (or they) exist, referentials which express the state of knowledge before launch of the new drug. The drug's profile (indication or contraindication, etc.), at the point when the marketing authorization is given, is purely theoretical. One must evaluate the real place of the drug under its real conditions of use (pragmatic trials, observable surveys). A new drugs' place in a therapeutic strategy can only be evaluated in the course of time unless a therapeutic revolution occurs. PMID- 10667113 TI - [Education and role of the toxicologist]. AB - Toxicology covers a very large scientific area, needing specialists in the main basic disciplines involved, and generalists with a large and practical overview. Academic education, consequently, should take this necessity into account. This education is presently satisfactory in France, even if some adjustments are necessary, for the specialists but is inadequate for the generalists, whereas an important need exists in industry. It should be emphasized that toxicology is a profession and consequently practical experience is essential, as well as continuing education. In view of this situation, an evaluation of toxicologists in the course of their professional activities is necessary. In the USA, this evaluation is devolved to the American Board of Toxicology and in Europe to the Eurotox Accreditation. In conclusion of the Round Table, 8 recommendations are proposed. PMID- 10667114 TI - [How can institutional structures make clinical research in France more operational?]. AB - The laws regulating the practice of clinical research in France, in particular the law of 20 December 1988, the so-called Huriet's law, constitute a major advance for medical progress. However, their implementation by administrative offices generates practical difficulties which impair the development of applied research in human beings. Beyond the laws themselves, it appears that our institutions are unprepared to optimize the conduct of such research. This round table sought to list the existing problems and to propose constructive solutions or objectives to be reached to optimize clinical research in France, with a view to improving French participation in international collaborative programmes, notably European ones. Evaluation of projects and practices, financial support and accounting, and some aspects of existing laws have been identified as the major sources of our difficulties. Harmonization and clarification of our procedures as well as improvement of training should be our primary objectives to achieve a higher level of medical, scientific, financial and administrative quality in the conduct of clinical research. Creation of a referential Web site, designed and updated by a central public organization, is an imperative step towards reaching these objectives. PMID- 10667115 TI - [Making the administration of dry oral forms of cytotoxic medications safer]. PMID- 10667116 TI - [Safe oral administration of etoposide: application to pediatric practice]. PMID- 10667117 TI - [Carbamazepine intolerance mimicking infectious mononucleosis: apropos of 3 cases]. PMID- 10667118 TI - [Lansoprazole hypersensitivity syndrome]. PMID- 10667119 TI - [Acute disseminated lupus erythematosus. Responsibility of Corenitec: apropos of a case]. PMID- 10667120 TI - [Latanoprost (Xalatan) and a systemic respiratory effect? Apropos of a case]. PMID- 10667121 TI - [Cardiotoxicity and immunomodulators: apropos of a case]. PMID- 10667122 TI - [Belief, science and morals of the majority]. PMID- 10667123 TI - [Electromagnetic effects of pacemaker-systems and the problem of the year 2000]. PMID- 10667124 TI - [Anxiety in genes and for genes]. PMID- 10667125 TI - [Pediatric day surgery of the respiratory tract--indications and prioritization]. AB - Ear, Nose and Throat (ENT) procedures are the most common types of surgery in children and include adenoidectomy, tonsillectomy, myringotomy, ventilation tube insertion or combinations of these. In order to study disease profile and routines for referral and treatment in outpatient otolaryngologic surgery, data were collected from 178 children operated consecutively during a six-week period in 1998. Median time from referral to surgery was less than four months. The majority of children operated for recurrent acute otitis media, tonsillitis or upper respiratory infections had suffered from the disease for 12 months or less. Obstructive symptoms were registered in 18% of these children. Most patients were referred to the hospital by specialists in otolaryngology or paediatric medicine. Surgery was more common in male than female-children, and median age at the time of surgery was 4.2 years. There was an equal distribution of middle ear and pharyngeal surgery. PMID- 10667126 TI - [Efficient learning with data from own practice--experiences from the SATS project]. AB - Between 1995 and 1998 the Norwegian Medical Association carried out a project to develop and assess a quality improvement tool for use in general practice (SATS). This method combines self-directed learning, documentation of practice and peer group support. SATS defined performance indicators for registration of practice by means of computerised patient records. Groups of four to ten general practitioners used their own consultation data as a basis for learning cycles. The practice evaluation indicates significant improvement in clinical work. Participating doctors found that having their own recorded data examined in a supportive peer environment was a major force for change. They reported satisfaction with the method, and expressed an interest in trying out new topics. However, the project demonstrated the need for simplification of terminology, further development of group process methods and computer software. There is also a need for strong local support of peer review groups. PMID- 10667127 TI - [Transformed migraine--chronic daily headache]. AB - Transformed migraine is probably a common cause of chronic daily headache. The International Headache Society system of diagnostic classification of headache classifies headaches, not patients, and pays no attention to the long-term evolution of the patient's headache. We support the suggestion made by Silberstein et al. that transformed migraine should be used as a diagnostic label in patients suffering from chronic daily headache with "migrainous features" and a history of migraine. Since 1994, intravenous treatment with dihydroergotamine (DHE) has been used for these patients. Among 16 patients suffering from chronic daily headache which were treated with this drug in 1996, 10 patients reported complete relief of headache at discharge from the hospital, but only four patients experienced complete relief from headache for more than two weeks. We think that the most important treatment for these patients are medicament withdrawal, information, help and support so that patients may cope with their headache. Dihydroergotamine can help patients going through a withdrawal regime. PMID- 10667128 TI - [Miniinvasive plate osteosynthesis of distal tibial fractures]. AB - The end result after open reduction and internal fixation in distal leg fractures is jeopardized by soft tissue complications. Functional conservative treatment of the same type of fractures often results in delayed and non-union, malunion or non-optimal functional result concerning ankle mobility. Mini-invasive plateosteosynthesis was performed in ten patients with sustained juxta or intra articular fractures of the distal tibia. In all patients, stable osteo-fixation could be achieved and soft tissue complications did not affect the final result in any of the patients. Mini-invasive plateosteosynthesis seems to be a good treatment alternative in extra- or intraarticular fractures of the distal leg. PMID- 10667129 TI - [Low molecular heparin in a pregnant women with heart valve prosthesis]. AB - Women with a prosthetic heart valve who wish to bear a child face a difficult choice. Continued medication with warfarin during pregnancy will offer her a reasonable good protection against thrombosis, but carries a considerable risk of teratogenic effects. Subcutaneously administered heparin is an alternative to warfarin. Standard (unfractionated) heparin has not offered sufficient protection against valve thrombosis, which may be a life-threatening complication. We were contacted by a woman who wished to become pregnant and who did not want to use warfarin if this implied a risk for the foetus. We followed a protocol using low molecular weight heparin in rather high doses. The dose was adjusted according to the results of blood test controls, which confirmed increased need for anticoagulation as the pregnancy evolved. Serial eccocardiography excluded valve thrombosis. The woman needed psychological support from her general practitioner. No complications occurred, and she gave birth to a normal child. We describe the various alternatives for anticoagulant treatment in pregnant women with mechanical heart valves. The choice of anticoagulant regimen and their consequences must be thoroughly discussed with the woman. PMID- 10667130 TI - [Myelomatosis and low level of vitamin B12]. AB - Many patients with multiple myeloma tend to have low serum cobalamin. The cause of this remains unclear. The important issue is whether cobalamin therapy should be used or not. We describe one case of megaloblastic erythropoiesis and multiple myeloma, and refer to some of the few studies describing the subject. Most of the patients with multiple myeloma are elderly, and the frequency of hypo- and achlorhydria is therefore increased. It has been demonstrated that cobalamin uptake and consumption is higher in myeloma cells than in normal bone marrow cells, and that cobalamin may be required for paraprotein synthesis. These facts may suggest that patients with multiple myeloma are more vulnerable to developing megaloblastic anemia than others. Our patient received cobalamin therapy in addition to cytostatic therapy for multiple myeloma without complications. However, we cannot exclude that cobalamin therapy may accelerate multiple myeloma; this should be considered when such therapy is given. However, accurate guidelines require more studies. PMID- 10667131 TI - [Why is dioxin harmful?]. AB - The scandal in Belgium last spring has drawn attention to the environmental hazards of dioxins. Previous production of pesticides and widespread combustion of organic material in the presence of chloride have lead to environmental accumulation of these toxicants, which more precisely are termed polychlorinated dibenzo-p-dioxins and dibenzofurans. Their very long biological half-lives in combination with detectable biological effects at very low concentrations have caused health concerns. Chloracne is the only well documented health effect in man, but there are experimental evidence for carcinogenic, teratogenic, reproductive and immunosuppressive effects. In this presentation we review current knowledge about the cellular effects of dioxins. Dioxins bind to and exert their effects through the cytoplasmic aryl hydrocarbon receptor, which acts as a transcription factor and regulates a number of cytokines and microsomal enzymes. Furthermore, dioxins interfere with hormonal signalling, and anti oestrogenic effects, vitamin A inhibition and thyroxin mimicry have been reported. Recently, effects on intracellular growth factor signalling have been demonstrated. Dioxins inhibit epidermal growth factor receptor, activate protein kinase C and other intracellular signal transducers, and activate transcription factors. As overall understanding of their cellular mechanisms of toxicity is lacking, we do not possess a complete basis for estimating the adverse health effects of this group of environmental toxicants. PMID- 10667132 TI - [Assessment of gestational age using ultrasound--can the method be improved?]. AB - In Norway, ultrasound measurement of the fetal biparietal diameter is used to determine the date of confinement according to Eik-Nes & Grottum's method. We aimed to evaluate the precision of this method. 8,029 women with singleton pregnancy and spontaneous vaginal delivery were arranged in groups according to gestational age at the time of the ultrasound examination. The precision of the biparietal diameter measurement for predicting the date of birth was determined for each group. An alternative method by Altman & Chitty was also tested on the population. Eik-Nes & Grottum's method predicts well the date of confinement if the biparietal diameter is measured at 17-20 weeks. Measurements at an earlier stage predict the date of birth with less confidence, particularly during completed gestational weeks 13-16, when the mean error reaches four days. Altman & Chitty's chart seemed to perform more evenly for the various gestational ages, but was systematically shifted by 3-4 days when used on our population. Eik-Nes & Grottum's chart for assessing gestational age should preferably not be used before 17 weeks of gestation. Introducing new charts based on a different population is not a good option. New charts based on a Norwegian population are needed. PMID- 10667133 TI - [Internet and the physician-patient relationship--from "thank you" to "why"?]. AB - The combination of modern information technology and an increasing awareness if consumer rights in the general population has lead to more demanding patients and new challenges for doctors. By using a questionnaire we explored the extent to which doctors feel that their clinical work is influenced by quality-demanding and well-informed patients, and how the internet influences the communication between doctor and patient. A representative sample of 1,276 Norwegian doctors doing clinical work responded to the survey. A great majority of the doctors expressed that "the informed patient" is becoming an integral part of their regular workday. Three out of four Norwegian doctors had experience with patients bringing internet information to the consultation setting. Most of these doctors found this natural and unobtrusive; a few felt it influenced the doctor-patient relationship in a negative way, while one out of every four doctors found meeting "the informed patient" a positive challenge. One out of seven doctors with e-mail access sporadically receives electronic mail from their patients, a few doctors get such messages often. So far Norwegian doctors are of the opinion that the new information technology has not introduced major changes or created unexpected difficulties in the doctor-patient relationship. PMID- 10667134 TI - [The use of Internet among Norwegian physicians]. AB - Access to the Internet increases rapidly, also among physicians. Little is known, however, of to what extent this tool is actually used among various groups of medical professionals, and how it compares to the use of more traditional ways of professional development such as reading of medical literature or participation in continuing medical education (CME) activities. In December of 1998, a 10% random sample of all Norwegian doctors, 1,646 in all, were mailed a questionnaire regarding their use of the Internet. 78% responded. 72% of Norwegian doctors had Internet access, 24% respectively had access either at work, at home or at both places. Doctors with access both at work and at home used the Internet significantly more often and found it of greater professional value than did the other groups. A smaller proportion of general practitioners than of other groups in the profession, had access. About half (48%) of Norwegian doctors use the Internet in a professional context. Research-oriented, male doctors, 30-49 years of age, indicated the highest activity on the net. Doctors using the Internet professionally had longer working hours, read more medical literature and participated more often in CME activities than did non-users. With its universal accessibility, the Internet has been seen as a major force in making medical knowledge available to all doctors. This is not yet the case; for the time being, it appears that the net widens the gap between doctors who actively seek new professional knowledge and those who do not. PMID- 10667135 TI - [Teleradiology--an effective agent for solidarity of health politics?]. PMID- 10667136 TI - [How can we justify scientific research?]. AB - The purpose and goal of scientific research have always been of concern to philosophers and ethicists. This concern is closely linked to the values that have justified scientific research. This paper takes a closer look at these values: professional personal fulfillment, social utility, and methodological norms such as value neutrality, openness and autonomy. These values originated in different historical epochs; however, they are still of great importance, e.g. in setting research priorities. PMID- 10667137 TI - [Medicine with a human face--a scientific philosophic reflection]. AB - The essay deals with the relationship between the scientific and humanistic social scientific aspects of medical concept formation and science. The first part focuses on methodological differences associated with the two forms of knowledge, emphasizing the distinction between quantitative and qualitative methods. The differences are traced back to type of phenomenon or object researched. On the basis of the special character of human beings and their illnesses it is then argued that scientific medicine necessarily will have to transcend the divide between the natural sciences on the one hand and the humanities and social sciences (the "human sciences") on the other. The unity in medicine is tied to its practice--the art of medicine as carried on in the clinical meeting between doctor and patient. PMID- 10667138 TI - [Reconsiliation, redemption and growth--a different perspective on the euthanasia debate]. PMID- 10667139 TI - [Psychiatrists' certifications in diffuse disorders]. PMID- 10667140 TI - [Ritual circumcision should be performed in private practice]. PMID- 10667141 TI - [About the feedback between medicine and culture]. PMID- 10667142 TI - [American health care]. PMID- 10667143 TI - [Image of physicians as professionals]. PMID- 10667144 TI - Shortage of supplies forces suspension of schools' BCG programme. PMID- 10667145 TI - [The infected artificial hip joint: possibilities, follow-up and results of treatment]. AB - Infections after total hip replacement are still a severe problem. With current therapies they can be managed successfully. In the present study we investigated 38 patients who were treated in our clinic because of infected total hip arthroplasties. All patients had a minimal follow-up of two years. The three main therapeutical forms are explained: the revision without removal of the prosthesis, the one-stage exchange and the two-stage exchange. 35 of the 38 patients could be healed within the observation period (from 1984-1996). Two patients died in the postoperative period from cardiopulmonary diseases. One patient could not be healed and has still an ongoing infection. The success rate was 92%, with good clinical results regarding pain, and walking ability. For the patient, the treatment of an infection can be very perturbing. With the new therapeutical forms the strain for the patient can be decreased. The type and duration of the therapy before definitive treatment has no influence on the success rate. In cases with a previous long-term therapy, a more complex treatment may be necessary. The treatment should be performed in an experienced clinic. The general practitioner plays an important part in the diagnosis and after care. An adequate treatment can be offered to nearly every patient with an infected total hip arthroplasty. This also applies to patients with long-term infection and chronic fistulation. PMID- 10667146 TI - [Swallowed dental prosthesis]. AB - A 66 year old man presented at the ambulatory with acute dysphagia. Inspectorial and radiologic examination revealed a swallowed dental prosthesis. The importance of the history and the clinical and radiological examination are discussed. PMID- 10667148 TI - 20th Annual meeting of the Society for Maternal-Fetal Medicine. Miami Beach, Florida, USA. January 31-February 5, 2000. Abstracts. PMID- 10667147 TI - [Secondary amenorrhea with hyperprolactinemia. 23-year-old Turkish patient, chief symptom: amenorrhea]. PMID- 10667149 TI - 10th International Hamburg Symposium on Tumor Markers. Hamburg, Germany, 5-7 December 1999. Abstracts. PMID- 10667150 TI - 78th General session of the International Association for Dental Research, in conjunction with the 29th annual meeting of the American Association for Dental Research and the 24th annual meeting of the Canadian Association for Dental Research. Washington, DC, USA. April 5-8, 2000. Abstracts. PMID- 10667151 TI - Cytologic changes in hepatocellular carcinoma after percutaneous acetic acid injection. Correlation with helical computed tomography findings. AB - OBJECTIVE: To illustrate the cytologic features of hepatocellular carcinoma (HCC) after percutaneous acetic acid injection (PAI) and to correlate the cytologic findings with helical computed tomography (CT) findings. STUDY DESIGN: The study included 30 patients with 37 HCC who had undergone PAI. Baseline cytomorphology of HCC was evaluated by needle aspiration in all cases. PAI under ultrasound guidance was done every three to seven days. Upon completion of PAI, fine needle aspiration cytology was performed and followed by helical CT within two weeks. The degeneration of HCC after PAI was classified into two grades. Grade 1 showed incomplete degeneration (99% of nuclear area); grade 2 showed complete degeneration or severe degeneration with cell debris or amorphous material only. The specimens were stained with Riu's method (Romanowsky system). RESULTS: The cytologic changes after PAI included decreased cell number, reduced cellular aggregation, degeneration of cytoplasm and nucleus, and eosinophilic or basophilic background in all tumors. In all the 37 tumors without enhancement on helical CT, grade 2 degeneration was detected. CONCLUSION: Our results reveal that grade 2 degeneration alone, demonstrated cytologically, could indicate almost complete necrosis of HCC after PAI, probably implying no need for booster PAI. PMID- 10667152 TI - Immunocytochemical staining of Kupffer and endothelial cells in fine needle aspiration cytology of hepatocellular carcinoma. AB - OBJECTIVE: To evaluate the significance of the presence of Kupffer and endothelial cells in distinguishing hepatocellular carcinoma (HCC) and adenocarcinoma (AC) on cytologic smears. STUDY DESIGN: Fine needle aspiration biopsies (FNABs) from 43 cases, 21 HCC and 22 AC (8 primary and 14 metastatic), were immunostained for Factor VIII and vimentin as markers for endothelial cells and Kupffer cells, respectively. Cytologic diagnosis was verified by histologic and/or clinical follow-up. RESULTS: Eighteen of the 21 cases (86%) of HCC and 11 (5 primary and 6 metastatic) of 22 cases (50%) of AC showed positive immunostaining for Factor VIII (P = .02). Vimentin immunostaining was positive in 55% of HCCs and 41% of ACs (P = .74). Forty-five percent of cases of HCC showed immunostaining for both Factor VIII and vimentin, while 22% of cases of AC showed immunostaining for both Factor VIII and vimentin (P = .18). CONCLUSION: Immunocytochemical identification of endothelial cells using Factor VIII may have important diagnostic value in separating HCC from adenocarcinomas in liver FNABs. The presence of Kupffer cells labeled with vimentin has no diagnostic significance in FNAB of these tumors. PMID- 10667153 TI - Cytologic detection of cervical and endometrial carcinoma with other genital tract involvement. AB - OBJECTIVE: To investigate the possibility of a correct cytologic diagnosis of cervical and endometrial carcinoma with other genital organ involvement. STUDY DESIGN: From uteri removed during hysterectomy due to cervical (33 cases) and endometrial (44 cases) cancer, samples were taken by cytobrush or spatula from the ectocervix, endocervix and endometrium of uteri opened longitudinally. Smears and cytosediments were stained by the Papanicolaou polychrome method. Moreover, acid beta-galactosidase activity was demonstrated in serial cytosediments by the indigogenic method of Lojda. From quenched tissue samples taken from the same sites as those for cytology, a series of cryostat sections was prepared and stained by hematoxylin and eosin or azure A, or subjected to the reaction for acid beta-galactosidase. RESULTS: In 17 of 33 patients with cervical cancer, the same type of cancer was also found in smears of the endocervix and endometrium. In six patients the type of cancer was different. Of 44 patients with endometrial cancer, 16 had an endocervical malignancy of the same type. In seven cases the type of cancer was different. The reaction for acid beta-galactosidase helped in the differentiation between squamous (negative reaction in cancer cells) and cylindrocellular (positive reaction) cancer in cytologic preparations. CONCLUSION: Before treatment, it is necessary to determine if there is involvement of the endocervix in endometrial cancer and of the endometrium in cervical cancer. Routine cytologic examination supplemented by the reaction for acid beta-galactosidase proved to be useful for this purpose. PMID- 10667154 TI - Cytologic diagnosis of axillary lymph node metastasis in breast cancer. AB - OBJECTIVE: To compare imprint cytology with histology as a method for rapid intraoperative diagnosis of axillary lymph node metastasis in breast cancer. STUDY DESIGN: We evaluated imprint cytology, comparing it with histopathology. A sample of 635 axillary lymph nodes was studied by imprint cytology using both Giemsa stain and hematoxylin-eosin. The results were compared with each other and with those of histopathologic examination. RESULTS: The Giemsa stain method, as compared to histopathology, had 94% accuracy, 97% sensitivity, 90% specificity and 94% positive prognostic value. The hematoxylin-eosin stain method was less accurate than the Giemsa stain method as compared to histopathology (accuracy 91%, sensitivity 96%, specificity 83% and positive prognostic value 92%). CONCLUSION: These data confirm the value of imprint cytology as a rapid, reliable method of intraoperative assessment of axillary lymph node metastasis in breast cancer. It results in better staging of the disease. It can be used intraoperatively, as an alternative to frozen section, if a pathology laboratory is not available, to exclude stage I patients from further treatment. PMID- 10667155 TI - Clinical impact of atypical squamous cells of undetermined significance. A cytohistologic comparison. AB - OBJECTIVE: To assess the percentage of squamous intraepithelial lesions (SILs) in the atypical squamous cells of undetermined significance (ASCUS) cytologic diagnosis. STUDY DESIGN: From January 1994 to December 1995, 421 cervical Pap smears with a diagnosis of ASCUS were followed with cervical biopsies within three months. The ASCUS cytologic diagnosis was correlated with the histologic findings and stratified according to age group, previous abnormal history and cell type of ASCUS (squamoid vs. metaplastic). RESULTS: Histologic diagnosis showed that of ASCUS diagnoses, 13% were normal, 34% were reactive, 4.8% were atypical, 43% were low grade SIL, 4% were high grade SIL, 1% were carcinoma in situ, and none were invasive lesions. The patients in the youngest group, up to 25 years, demonstrated the highest percentage of SIL. Patients with a previous abnormal gynecologic history showed a higher percentage of SIL than those without an abnormal history. SILs were observed in 51.5% of squamoid ASCUS and 36.5% of metaplastic ASCUS. CONCLUSION: Forty-eight percent of females having an ASCUS diagnosis on Pap smears had SIL and thus a preneoplastic lesion. The highest percentage of SIL was found in females 25 years and younger. Our findings suggest that an ASCUS diagnosis warrants ongoing follow-up. PMID- 10667156 TI - p53 immunohistochemistry for distinguishing reactive mesothelium from low grade ovarian carcinoma. AB - OBJECTIVE: To determine the utility of immunohistochemical staining for p53 in cell block material for distinguishing reactive mesothelium from borderline or low grade ovarian carcinoma. STUDY DESIGN: Paraffin-embedded cell blocks from paracentesis and pelvic wash fluid of 44 cases of ovarian carcinoma and 20 cases containing only reactive mesothelium were immunostained for p53 using monoclonal antibody DO-7. Tumor grades ranged from borderline to high grade and were serous papillary (33), clear cell (3), mucinous (2), endometrioid (2), mixed serous papillary/clear cell (3) and undifferentiated (1). The three authors independently evaluated the staining, including estimation of the percentage and intensity of positive nuclear staining. RESULTS: A separation of positive from negative cases was seen when staining intensity was considered the critical parameter; moderate to strong staining was considered truly positive. Seventy three percent (8/11) of borderline tumors, 80% (8/10) of low grade tumors and 65% (15/23) of intermediate to high grade tumors showed moderate to strong positivity. Percentage of staining was a less-reliable parameter as 25% of negative cases were positive by this assessment. CONCLUSION: p53 Immunohistochemistry, using monoclonal antibody DO-7 combined with standard morphologic evaluation, may be useful in distinguishing benign reactive mesothelium from borderline or low grade ovarian carcinoma. PMID- 10667157 TI - Characteristics of high grade dyskaryotic cervical smears likely to be missed on rapid rescreening. AB - OBJECTIVE: To determine if there is a type of high grade dyskaryotic cervical smear that is likely to be missed on rapid rescreening. STUDY DESIGN: Fifty high grade dyskaryotic smears that had originally been incorrectly reported as negative (FN) were admixed with 100 true negative smears. Each smear in the set was rapidly reviewed at least 40 times. The FN smears that were picked out on > 50% of screenings were compared with those that were passed as unremarkable on > 50% of screenings for features of the dyskaryotic cell population. RESULTS: Significant differences between the two types of FN smear were present in five aspects of the dyskaryotic cell population. A FN smear is more likely to be missed on rapid rescreening than to be selected for review if it has few dyskaryotic cells; if the dyskaryotic cells are small, with pale nuclei; and if they are scattered singly rather than in groups or syncytia. CONCLUSION: A type of severely dyskaryotic smear is likely to evade rapid rescreening as well as routine screening. This suggests that even when rapid rescreening is used as a quality assurance measure, the "zero-error standard" is unlikely to be attained. PMID- 10667158 TI - Cytohistologic correlation between AGUS and biopsy-detected lesions in postmenopausal women. AB - OBJECTIVE: To evaluate histologic findings in patients aged 50 and older whose cervical smears revealed atypical glandular cells of undetermined significance (AGUS). STUDY DESIGN: Computerized records spanning a four-year period were retrospectively analyzed. Thirty patients over age 50 had cervical smears interpreted as AGUS and had follow-up biopsies within 12 months following the abnormal smear. The most important histologic diagnosis from the biopsy specimens was correlated with the subcategory of the cervical smear. RESULTS: Five smears interpreted as AGUS, favor reactive, revealed abnormal histology in four cases: three endometrial polyps and one squamous carcinoma. Two smears interpreted as AGUS, favor dysplasia, revealed squamous intraepithelial lesions on biopsy in both cases. Seventeen smears interpreted as AGUS, favor endometrial cells, revealed abnormal histology in 13 cases: 1 endocervical polyp, 6 endometrial polyps, 3 endometrial hyperplasias and 3 adenomyosis. Six patients with smears interpreted as AGUS, unclassifiable, revealed abnormal histology in five cases: two endocervical polyps, one endometrial polyp, one endometrial carcinoma and one ovarian carcinoma. CONCLUSION: The presence of AGUS in cervical smears from women over 50 was highly predictive of abnormal lesions detected by histologic examination. Although three cancers were detected on histologic follow-up, the most common lesions detected were endometrial polyps. PMID- 10667159 TI - Comparison of ThinPrep and conventional preparations on fine needle aspiration cytology material. AB - OBJECTIVE: To compare the various cytologic features on ThinPrep 2000 (TP) (Cytyc Corporation, Marlborough, Massachusetts, U.S.A.) and conventional preparation (CP) specimens from fine needle aspiration cytology (FNAC) material by a semiquantitative scoring system. STUDY DESIGN: In this prospective study a total of 71 consecutive cases were included. In each case, two passes were performed. The first pass was used for conventional preparations, with direct smears made and fixed immediately in 95% alcohol for Papanicolaou stain. For TP preparation a second pass produced material for processing in the ThinPrep 2000. The TP and CP slides were studied independently by two observers and representative slides of CP and TP compared for cellularity, background blood and necrotic cell debris, cell architecture, informative background, presence of monolayer cells, and nuclear and cytoplasmic details by a semiquantitative scoring system. Statistical analysis was performed by Wilcoxon's signed rank test on an SPSS program (Chicago, Illinois, U.S.A.). RESULTS: TP preparations contained adequate diagnostic cells in all cases and were tangibly superior to CP preparations concerning monolayer cells, absence of blood and necrosis, and preservation of nuclear and cytoplasmic detail (statistically significant, Wilcoxon's signed rank test, P < .000). CONCLUSION: TP preparations are superior to conventional preparations with regard to clear background, monolayer cell preparation and cell preservation. It is easier and less time consuming to screen and interpret TP preparations because the cells are limited to smaller areas on clear backgrounds, with excellent cellular preservation. However, TP preparations are more expensive than CP and require some experience for interpretation. PMID- 10667160 TI - Identification of amylase crystalloids in cystic lesions of the parotid gland. AB - OBJECTIVE: To identify alpha-amylase crystalloid formations in parotid specimens obtained by fine needle aspiration. STUDY DESIGN: The study concerned three cases of sialadenitis with crystalloid formation observed between 1993 and 1998. In one of these cases, transmission electron microscopy, mass spectrometry and measurement of amylase activity were used to characterize the nature of the crystalloids. RESULTS: Light microscopy revealed the same crystalloid structure in all three cases. In one case, where the material was saved, a biochemical method made it possible to reveal high amylase activity, while protein electrophoresis and mass spectrometry were used to identify salivary alpha amylase. CONCLUSION: Crystalloids of salivary alpha-amylase can be identified by May-Grunwald-Giemsa and Papanicolaou stain and can be rapidly confirmed through determination of amylase activity. PMID- 10667161 TI - Imprint cytology of core needle biopsy specimens of breast lesions. A rapid approach to detecting malignancies, with comparison of cytologic and histopathologic analyses of 173 cases. AB - OBJECTIVE: To investigate whether imprint cytology of core needle biopsy (CNB) specimens from breast lesions is a useful method of rapidly obtaining additional diagnostic information and potentially can be used to reduce the number of biopsies needed. STUDY DESIGN: Cytologic analysis was performed on 173 breast lesions and compared with their histopathologic diagnoses (143 malignant and 30 benign). For imprint cytology, one CNB specimen was rolled between two slides and stained with Diff-Quik and Papanicolaou stain. RESULTS: The diagnostic overall accuracy of Diff-Quik stain (Papanicolaou stain) was 95.4% (95.9%), with a sensitivity of 96.5% (97.2%), specificity of 90% (90%), positive predictive value of 97.8% (97.8%) and negative predictive value of 84.3% (87.0%). There was no statistically significant difference between the stains. Histopathologic analysis had an overall accuracy of 97.7%, with a sensitivity of 97.2%, specificity and positive predictive value of 100% and a negative predictive value of 88.2%. CONCLUSION: Imprint cytology of CNBs is a sensitive method of detecting malignancies in breast tumors. Diff-Quik is a rapid and reliable approach that can reduce the number of biopsies. Inadequate and suspicious cases should be evaluated based on complementary diagnostic procedures for breast lesions. PMID- 10667162 TI - Molluscum contagiosum. A case report with fine needle aspiration cytologic diagnosis and ultrastructural features. AB - BACKGROUND: Cytomorphologic and ultrastructural features of molluscum contagiosum, a rare skin lesion of viral etiology, are presented. CASE: A 4-month old female was referred for fine needle aspiration cytology of papules over the back and chest wall. A Giemsa-stained preparation of whitish material aspirated from the chest wall nodule showed numerous large, intracytoplasmic, basophilic bodies that pushed the host cell nucleus to the periphery, giving a signet-ring appearance to a few cells. A cytologic diagnosis of molluscum contagiosum was suggested. On electron microscopy numerous intracytoplasmic viral particles were demonstrated, thus confirming the cytologic diagnosis. CONCLUSION: In clinically unsuspected cases, the cytologic diagnosis of molluscum contagiosum can be suggested by demonstrating pathognomonic molluscum bodies in aspirated material. PMID- 10667163 TI - Fine needle aspiration cytology in systemic lupus erythematosus lymphadenopathy. A case report. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown etiology characterized by inflammation in various organ systems, including lymph nodes, due to the production of antinuclear antibodies. The onset of disease is between ages 13 and 40 years, with a female preponderance. CASE: A 30-year-old female presented with right cervical lymphadenopathy and gave a history of intermittent fever and swollen joints of 2.5 years' duration. The patient was on intermittent corticosteroids. With a suggestion of tuberculous lymphadenitis, the patient underwent fine needle aspiration (FNA). The diagnosis of lupus adenopathy was established by FNA of enlarged right cervical lymph nodes. Smears showed predominantly typical and atypical immunoblasts, plasma cells, occasional Reed-Sternberg-like cells and dispersed hematoxylin bodies. Smears were negative for acid-fast bacilli. CONCLUSION: When SLE patients develop lymphadenopathy, FNA cytology helps differentiate lupus adenopathy from infectious conditions, such as tuberculous adenitis, and from Kikuchi's lymphadenitis. PMID- 10667164 TI - Diagnosis of malignant mesothelioma by fine needle aspiration of a cervical lymph node. A case report. AB - BACKGROUND: Clinically documented distant metastases are rare in mesothelioma and tend to occur late in the course of the disease, well after the diagnosis has been made. In this instance, diagnosis was not made until a metastatic deposit was identified microscopically in the enlarged lymph node. CASE: A 65-year-old male with no definite history of occupational asbestos exposure presented with chest pain, pleural effusion and supraclavicular lymphadenopathy. Cytologic examination of material obtained by fine needle aspiration from his cervical lymph node revealed malignant mesothelioma. This was confirmed on histology. CONCLUSION: This was a particularly rare presentation and, as far as we are aware, was the first case in which mesothelioma was diagnosed by fine needle aspiration of a cervical lymph node. It serves to remind the pathologist that when confronted with a lymph node involved by tumor, the possibility of mesothelioma should be included in the differential diagnosis. The case also demonstrates the usefulness of fine needle aspiration in the diagnosis of metastatic tumor. PMID- 10667165 TI - North American paragonimiasis. A case report. AB - BACKGROUND: Paragonimiasis is a parasitic infection with a predilection for pulmonary involvement. Paragonimus species occur throughout the world and exist in nature in a snail-crustacean-mammalian life cycle. Human disease is most frequently encountered in cultures that ingest raw or undercooked crustaceans. North American paragonimiasis, caused by an endemic Paragonimus species, Paragonimus kellicotti, predominantly causes disease in carnivorous and omnivorous animals but may cause human disease if the intermediate host, the crayfish, is ingested raw or undercooked. CASE: A previously healthy, 21-year-old male was infected with P kellicotti and developed parasitic hemoptysis. The disease was contracted through the ingestion of local, undercooked crayfish. Diagnosis was established through the morphologic examination of eggs in the cytologic preparation of bronchioalveolar lavage fluid. The patient was successfully treated with praziquantel and recovered without incident. CONCLUSION: Paragonimiasis is a cause of parasitic hemoptysis worldwide. Paragonimiasis is infrequently encountered in North America and is usually not considered in the differential diagnosis of hemoptysis unless specific risk factors are known. The cytologist or cytopathologist, therefore, may be the first to encounter the diagnostic eggs and should be familiar with this disease. PMID- 10667166 TI - Granular cell ameloblastoma of the jaw. A report of two cases with fine needle aspiration cytology. AB - BACKGROUND: Fine needle aspiration cytology (FNAC) of jaw tumors has not been studied extensively. Ameloblastomas are jaw tumors that show a wide morphologic spectrum and thus may pose some diagnostic difficulties. Of the many types, granular cell ameloblastoma (GCA) is an uncommon variant that possesses distinctive features. To the best of our knowledge, there have been no previous reports on the cytologic findings of GCA. We present two cases diagnosed by FNAC. CASES: Two cases of GCA were diagnosed on cytology and subsequently confirmed on histology. Both patients presented with a large, lytic jaw tumor. FNAC smears showed characteristic granular cells along with spindle and basaloid cells. CONCLUSION: Although GCAs are rare tumors, they possess distinctive features that permit an accurate diagnosis, provided that this entity is kept in mind. This tumor has to be differentiated from cystic odontogenic lesions, epulis and granular cell myoblastoma. An accurate preoperative diagnosis also helps the surgeon to plan more extensive surgery as these tumors show a great propensity for malignant change and metastases. PMID- 10667167 TI - Macrophagelike vacuolated renal tubular cells in the urine of a male with osmotic nephrosis associated with intravenous immunoglobulin therapy. A case report. AB - BACKGROUND: Osmotic nephrosis is a form of renal tubular injury that has been found in patients treated with intravenous immunoglobulin (IVIG). CASE: A 46-year old male who had two courses of chemotherapy for acute myelogenous leukemia was found to have refractory thrombocytopenia. After IVIG (Sandoglobulin 12%, Novartis) administration (1 g/kg) for five consecutive days, the patient became oliguric and eventually anuric on the fifth dose. Hemodialysis was initiated, and urine production was noted on day 2 of hospitalization. Routine cytologic examination of fresh, voided urine showed numerous macrophagelike, bland epithelial cells with abundant, multivacuolated cytoplasm. Cytokeratin immunostain revealed positivity, thus confirming the epithelial origin of these cells. CONCLUSION: To our knowledge, this is the first such case reported in the English-language cytology literature. Awareness of a patient's clinical history may be helpful in avoiding an incorrect diagnosis. Urine cytology may be useful in obtaining an early diagnosis of osmotic nephrosis in patients receiving high dose IVIG therapy that may eliminate the need for a renal biopsy. PMID- 10667168 TI - Plasmacytoma of the breast. A report of two cases diagnosed by aspiration biopsy. AB - BACKGROUND: Extramedullary plasmacytoma of the breast is an uncommon neoplasm, occurring either as a solitary tumor or as evidence of disseminated multiple myeloma. CASE: Two cases of plasmacytoma of the breast were diagnosed by fine needle aspiration cytology. Aspiration smears showed a dispersed population of plasmacytoid cells with eccentric nuclei, abundant cytoplasm and the characteristic paranuclear hof. CONCLUSION: The clinical, cytologic and immunophenotypic features of plasmacytoma are characteristic, and the importance of distinguishing these neoplasms from primary mammary tumors is important to avoid unnecessary surgery. PMID- 10667169 TI - Retroplasia of bronchial columnar cells mimicking intranuclear viral inclusions. PMID- 10667170 TI - Scraping cytology in the diagnosis of malignant squamous neoplasms of the skin. PMID- 10667171 TI - Pleural effusion as an initial manifestation of multiple myeloma. PMID- 10667172 TI - Fine needle aspiration cytology of lipoblastoma. PMID- 10667173 TI - Imprint cytology of an intrapulmonary lymph node. PMID- 10667174 TI - Microbes without borders: infectious disease, public health, and the journal. PMID- 10667175 TI - What's housing got to do with it? PMID- 10667176 TI - The microbial menace, then and now. PMID- 10667177 TI - The Occupational Safety and Health Administration and the public health model. PMID- 10667178 TI - Thinking about vaginal microbicide testing. AB - A vaginal microbicide could slow the spread of HIV. To date, volunteers in placebo-controlled trials of candidate microbicides have been counseled to use condoms. This does not reduce the number of volunteers exposed to possible risk, but it shifts the allotment of risk from those conducting the trial to those women who may be least able to make autonomous decisions. Alternative ways of meeting the obligation to offer volunteers active benefits are explored. Counseling the use of condoms prolongs clinical trials and could cause tens of thousands of otherwise avoidable deaths. PMID- 10667179 TI - The making of a germ panic, then and now. AB - Over the last 2 decades, a heightened interest in germs has been evident in many aspects of American popular culture, including news coverage, advertisements, and entertainment media. Although clearly a response to the AIDS epidemic and other recent disease outbreaks, current obsessions with germs have some striking parallels with a similar period of intense anxiety about disease germs that occurred between 1900 and 1940. A comparison of these 2 periods of germ "panic" suggests some of the long-term cultural trends that contributed to their making. Both germ panics reflected anxieties about societal incorporation, associated with expanding markets, transportation networks, and mass immigration. They were also shaped by new trends in public health education, journalism, advertising, and entertainment media. In comparison to the first germ panic, the current discourse about the "revenge of the superbugs" is considerably more pessimistic because of increasing worries about the environment, suspicions of governmental authority, and distrust of expert knowledge. Yet, as popular anxieties about infectious disease have increased, public health scientists have been attracting favorable coverage in their role as "medical detectives" on the trail of the "killer germ." PMID- 10667180 TI - Immunization and the American way: 4 childhood vaccines. AB - Childhood immunization constitutes one of the great success stories of American public health in the 20th century. This essay provides a historical examination of this topic through 4 particularly important examples: diphtheria, pertussis, polio, and measles. Each case study illustrates how new vaccines have posed unique challenges related to basic science, clinical trial methodology, medical ethics, and public acceptance. A brief comparison of each story to the experience of Great Britain, however, suggests an underlying unity connecting all 4 examples. Whereas the British led the way in introducing formal clinical trial methodology in the field of immunization development, the Americans excelled in the rapid translation of laboratory knowledge into strategies suitable for mass application. Although this distinction appears to have diminished in recent years, it offers insight into the sources of creativity underlying American vaccine development and the corresponding difficulties sometimes created for utilizing vaccines fruits rationally. PMID- 10667181 TI - Violence victimization after HIV infection in a US probability sample of adult patients in primary care. AB - OBJECTIVES: This study estimated the proportion of HIV-infected adults who have been assaulted by a partner or someone important to them since their HIV diagnosis and the extent to which they reported HIV-seropositive status as a cause of the violence. METHODS: Study participants were from a nationally representative probability sample of 2864 HIV-infected adults who were receiving medical care and were enrolled in the HIV Costs and Service Utilization Study. All interviews (91% in person, 9% by telephone) were conducted with computer assisted personal interviewing instruments. Interviews began in January 1996 and ended 15 months later. RESULTS: Overall, 20.5% of the women, 11.5% of the men who reported having sex with men, and 7.5% of the heterosexual men reported physical harm since diagnosis, of whom nearly half reported HIV-seropositive status as a cause of violent episodes. CONCLUSIONS: HIV-related care is an appropriate setting for routine assessment of violence. Programs to cross-train staff in antiviolence agencies and HIV care facilities need to be developed for men and women with HIV infection. PMID- 10667182 TI - A community-level HIV prevention intervention for inner-city women: results of the women and infants demonstration projects. AB - OBJECTIVES: This study examined the effects of a multisite community-level HIV prevention intervention on women's condom-use behaviors. METHODS: The theory based behavioral intervention was implemented with low-income, primarily African American women in 4 urban communities. It was evaluated with data from pre- and postintervention cross-sectional surveys in matched intervention and comparison communities. RESULTS: At baseline, 68% of the women had no intention of using condoms with their main partners and 70% were not using condoms consistently with other partners. After 2 years of intervention activities, increases in rates of talking with main partners about condoms were significantly larger in intervention communities than in comparison communities (P = .03). Intervention communities also had significant increases in the proportion of women who had tried to get their main partners to use condoms (P = .01). The trends for condom use with other partners were similar but nonsignificant. CONCLUSIONS: Many women at risk for HIV infection are still not using condoms. Community-level interventions may be an effective way to reach large numbers of women and change their condom-use behaviors, particularly their behaviors with regard to communication with main sex partners. PMID- 10667183 TI - Mortality from invasive pneumococcal pneumonia in the era of antibiotic resistance, 1995-1997. AB - OBJECTIVES: This study examined epidemiologic factors affecting mortality from pneumococcal pneumonia in 1995 through 1997. METHODS: Persons residing in a surveillance area who had community-acquired pneumonia requiring hospitalization and Streptococcus pneumoniae isolated from a sterile site were included in the analysis. Factors affecting mortality were evaluated in univariate and multivariate analyses. The number of deaths from pneumococcal pneumonia requiring hospitalization in the United States in 1996 was estimated. RESULTS: Of 5837 cases, 12% were fatal. Increased mortality was associated with older age, underlying disease. Asian race, and residence in Toronto/Peel, Ontario. When these factors were controlled for, increased mortality was not associated with resistance to penicillin or cefotaxime. However, when deaths during the first 4 hospital days were excluded, mortality was significantly associated with penicillin minimum inhibitory concentrations of 4.0 or higher and cefotaxime minimum inhibitory concentrations of 2.0 or higher. In 1996, about 7000 to 12,500 deaths occurred in the United States from pneumococcal pneumonia requiring hospitalization. CONCLUSIONS: Older age and underlying disease remain the most important factors influencing death from pneumococcal pneumonia. Mortality was not elevated in most infections with beta-lactam-resistant pneumococci. PMID- 10667184 TI - "Broken windows" and the risk of gonorrhea. AB - OBJECTIVES: We examined the relationships between neighborhood conditions and gonorrhea. METHODS: We assessed 55 block groups by rating housing and street conditions. We mapped all cases of gonorrhea between 1994 and 1996 and calculated aggregated case rates by block group. We obtained public school inspection reports and assigned findings to the block groups served by the neighborhood schools. A "broken windows" index measured housing quality, abandoned cars, graffiti, trash, and public school deterioration. Using data from the 1990 census and 1995 updates, we determined the association between "broken windows," demographic characteristics, and gonorrhea rates. RESULTS: The broken windows index explained more of the variance in gonorrhea rates than did a poverty index measuring income, unemployment, and low education. In high-poverty neighborhoods, block groups with high broken windows scores had significantly higher gonorrhea rates than block groups with low broken windows scores (46.6 per 1000 vs 25.8 per 1000; P < .001). CONCLUSIONS: The robust association of deteriorated physical conditions of local neighborhoods with gonorrhea rates, independent of poverty, merits an intervention trial to test whether the environment has a causal role in influencing high-risk sexual behaviors. PMID- 10667185 TI - Effectiveness of an intervention promoting the female condom to patients at sexually transmitted disease clinics. AB - OBJECTIVES: This study evaluated a behavioral intervention designed to promote female condoms and reduce unprotected sex among women at high risk for acquiring sexually transmitted diseases (STDs). METHODS: The effect of the intervention on barrier use was evaluated with a pretest-posttest design with 1159 female STD clinic patients. RESULTS: Among participants with follow-up data, 79% used the female condom at least once and often multiple times. More than one third of those who completed the study used female condoms throughout follow-up. Use of barrier protection increased significantly after the intervention, and high use was maintained during a 6-month follow-up. To account for attrition, the use of protection by all subjects was projected under 3 conservative assumptions. The initial visit and termination visit projections suggest that use increased sharply after the intervention and declined during follow-up but remained elevated compared with the baseline. CONCLUSIONS: Many clients of public STD clinics will try, and some will continue, to use female condoms when they are promoted positively and when women are trained to use them correctly and to promote them to their partners. A behavioral intervention that promotes both female and male condoms can increase barrier use. PMID- 10667186 TI - Factors associated with tympanostomy tube insertion among preschool-aged children in the United States. AB - OBJECTIVES: Recurrent and persistent otitis media is often treated by tympanostomy tube insertion to ventilate the middle ear and restore hearing. This study examined the factors that predict which children are most likely to receive tympanostomy tubes through 3 years of age. METHODS: Multiple logistic regression was conducted on data from a nationally representative sample of children (N = 8285). RESULTS: By 3 years of age, 6.8% of US children had tubes inserted. Logistic regression indicated that after control for number of ear infections, children without any gaps in health insurance, who attended a day-care center, who were White, whose birth-weight was less than 1500 g, and who lived in the Midwest or South were significantly more likely to have tympanostomy tubes. CONCLUSIONS: These data suggest that differences exist as to who receives tubes. Of particular concern are differences by race/ethnicity and continuity of health insurance coverage. With expansions in health care coverage to larger proportions of uninsured children, it will be important to monitor these programs to ensure that all children who may need tympanostomy tubes have access to them. PMID- 10667187 TI - Relationships between obesity and DSM-IV major depressive disorder, suicide ideation, and suicide attempts: results from a general population study. AB - OBJECTIVES: This study sought to test the relationships between relative body weight and clinical depression, suicide ideation, and suicide attempts in an adult US general population sample. METHODS: Respondents were 40,086 African American and White participants interviewed in a national survey. Outcome measures were past-year major depression, suicide ideation, and suicide attempts diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. The primary predictor was relative body weight, treated both continuously (i.e., body mass index [BMI]) and categorically in logistic regression analyses. Covariates included age, income and education, disease status, and drug and alcohol use. RESULTS: Relative body weight was associated with major depression, suicide attempts, and suicide ideation, although relationships were different for men and women. Among women, increased BMI was associated with both major depression and suicide ideation. Among men, lower BMI was associated with major depression, suicide attempts, and suicide ideation. There were no racial differences. CONCLUSIONS: Differences in BMI, or weight status, were associated with the probability of past-year major depression, suicide attempts, and suicide ideation. Longitudinal studies are needed to differentiate the causal pathways and mechanisms linking physical and psychiatric conditions. PMID- 10667188 TI - Risk factors for homelessness among indigent urban adults with no history of psychotic illness: a case-control study. AB - OBJECTIVES: This study identified risk factors for homelessness among indigent urban adults without dependent children and with no history of psychotic illness. METHODS: We conducted a matched case-control study, stratified by sex, of 200 newly homeless men and women and 200 indigent men and women with no history of homelessness. Newly homeless case subjects were recruited from shelter assessment centers in New York City. Never-homeless control subjects, selected from public assistance centers, were single adults applying for home relief. Control subjects were matched with case subjects according to ethnicity, age, and sex. Trained interviewers employed standardized research instruments to probe 3 domains of risk factors: symptom severity and substance use disorder, family support and functioning, and prior use of services. RESULTS: Significant interaction effects by sex were present for symptom severity, heroin use disorder, and prior service use. Greater numbers of the homeless of both sexes lacked a high school diploma and had less income from all sources, including from their families, than of the never homeless. CONCLUSIONS: Newly homeless men and women with no history of psychotic illness differed from their never-homeless counterparts in the 3 domains investigated, but socioeconomic factors were also important. PMID- 10667189 TI - Patterns and correlates of physical activity among US women 40 years and older. AB - OBJECTIVES: This study describes the patterns of physical activity among minority women by using a variety of definitions and determines sociodemographic and behavioral correlates of physical activity in this population. METHODS: A cross sectional study was conducted in 1996 and 1997 among US women 40 years and older (n = 2912) of the following racial/ethnic groups: African American, American Indian/Alaskan Native, Hispanic, and White. RESULTS: Physical activity was lowest among African Americans and American Indians/Alaskan Natives (adjusted odds ratios [ORs] for no leisure-time activity were 1.35 [95% confidence interval (CI) = 1.08, 1.68] and 1.65 [95% CI = 1.33, 2.06], respectively). A much higher proportion of women were classified as being physically active when occupational activity rather than more traditional assessments of leisure activity were used to determine level of physical activity. On the basis of a composite definition of physical activity, 72% of respondents reported being physically active. Women living in rural regions (OR = 1.33; 95% CI = 1.12, 1.58) were more likely than urban inhabitants to be completely inactive during leisure time. CONCLUSIONS: Minority women are among the least active subgroups in American society, although not all groups are less active than White women when all domains of physical activity are taken into account. PMID- 10667190 TI - Prevalence and risk factors of drug-resistant tuberculosis along the Mexico-Texas border. AB - OBJECTIVES: This study determined factors associated with drug resistance among 3496 patients with tuberculosis who resided in Texas counties along the Mexican border. METHODS: Univariate and logistic regression analyses were performed to identify risk factors associated with drug resistance. RESULTS: Among patients with a history of previous tuberculosis, being 19 years or younger was the only factor associated with multiple drug resistance. Female sex, being 20 to 39 years of age, and foreign birth were risk factors for resistance among patients with no history of previous tuberculosis. CONCLUSIONS: Factors contributing to drug resistance among Hispanic tuberculosis patients along the Texas-Mexico border may differ from those among other populations in the United States. PMID- 10667191 TI - Implementation of guidelines for HIV counseling and voluntary HIV testing of pregnant women. AB - OBJECTIVES: This study assessed HIV counseling and testing among pregnant women. METHODS: A survey was administered to 9115 women who gave birth at 66 Chicago area hospitals in 1997 and 1998. RESULTS: Fifty-eight percent of the women received HIV counseling, and 65% were offered testing. Fifty-six percent were tested for HIV. Among the women tested, 88% were given their test result. Women were more likely to be tested if they received HIV counseling and were more likely to be offered testing if they received such counseling. CONCLUSIONS: Rates of HIV counseling for, and offers of testing to, pregnant women need to be increased. PMID- 10667192 TI - Diminishing educational differences in breast cancer mortality among Finnish women: a register-based 25-year follow-up. AB - OBJECTIVES: This study examined trends in breast cancer mortality by education, age, and birth cohort. METHODS: Census records of Finnish women 35 years and older were linked with death records for 1971 through 1995. RESULTS: Excess breast cancer mortality of more-educated women has declined rapidly, mainly because of increasing mortality among less-educated women and stable or decreasing mortality among more-educated 35- to 64-year-old women. During the 1990s, mortality among more-educated 50- to 64-year-old women declined particularly fast. CONCLUSIONS: The causes of declining differences by education in breast cancer mortality are difficult to verify, but they may be due in part to narrowing differences in reproductive behavior among the younger birth cohorts and to a period effect possibly associated with the introduction of breast cancer screening in the late 1980s. PMID- 10667193 TI - Increasing trends in the use of breast-conserving surgery in California. AB - OBJECTIVES: The purpose of this study was to determine temporal trends in breast conserving surgery in California from 1988 through 1995. METHODS: Logistic regression was used to analyze data on 104,466 cases of early-stage breast cancer reported to the California Cancer Registry. RESULTS: A monotonically increasing trend in breast-conserving surgery was detected after adjustment for age, race/ethnicity, stage at diagnosis, and neighborhood education level. Breast conserving surgery increased at similar rates among all racial/ethnic groups. Older age, Asian or Hispanic race/ethnicity, late-stage diagnosis, and residence in an undereducated neighborhood were factors associated with lower use of breast conserving surgery. CONCLUSIONS: Although disparities are evident, use of breast conserving surgery increased steadily in all groups examined in this study. PMID- 10667194 TI - Acceptability and feasibility of urine screening for Chlamydia and gonorrhea in community organizations: perspectives from Denver and St Louis. PMID- 10667195 TI - Need for HIV/AIDS early identification and preventive measures among middle-aged and elderly women. PMID- 10667196 TI - Re McKinlay and Marceau's "Tale of 3 tails". PMID- 10667197 TI - Educating public health professionals. PMID- 10667198 TI - Telomerase. A target for anticancer therapy. AB - Telomerase is absent in most normal tissues, but is abnormally reactivated in all major cancer types. Telomerase enables tumor cells to maintain telomere length, allowing indefinite replicative capacity. Albeit not sufficient in itself to induce neoplasia, telomerase is believed to be necessary for cancer cells to grow without limit. The presence of telomerase has been detected in virtually all cancer types including the most prevalent cancers of the prostate, breast, lung, colon, bladder, uterus, ovary, and pancreas as well as in lymphomas, leukemias, and melanomas. In addition, data from cancer patients indicate that telomerase levels correlate with clinical outcome in neuroblastomas, leukemias, and prostate, gastric, and breast cancers. Studies using an antisense to the human telomerase RNA component demonstrate that telomerase in human tumor lines can be blocked ex vivo. In these experiments, telomerase inhibition led to telomere shortening and cancer cell death, validating telomerase as a target for anticancer therapy. Telomerase is a uniquely appealing target for drug discovery because its dichotomic expression in normal versus cancer cells suggests that no serious side effects would result from a treatment abrogating telomerase activity. A variety of approaches to telomerase inhibition are being investigated and are discussed. PMID- 10667199 TI - A paradigm for cancer treatment using the retinoblastoma gene in a mouse model. AB - Discovery of tumor suppressor genes has provided a rational approach to cancer prevention and treatment. Loss of retinoblastoma susceptibility gene (Rb) function is a rate-limiting event in the development of human and mouse cancers. Establishment of animal models of cancer associated with Rb deficiency allowed us to develop and test long-awaited approaches to genetic correction for treating tumors in vivo. Recent studies demonstrated that (1) prevention of carcinogenesis is achieved by correction of gene copy number in Rb+/- mice, and (2) reconstitution of Rb gene functions is sufficient for suppression of neoplasia in immunocompetent mice. These results fulfill a promise of cancer treatment by reconstitution of tumor suppressor function. PMID- 10667200 TI - Trichostatin and leptomycin. Inhibition of histone deacetylation and signal dependent nuclear export. AB - Trichostatin A (TSA), an inhibitor of the eukaryotic cell cycle and an inducer of morphological reversion of transformed cells, inhibits histone deacetylase (HDAC) at nanomolar concentrations. Recently, trapoxin, oxamflatin, and FR901228, antitumor agents structurally unrelated to TSA, were found to be potent HDAC inhibitors. These inhibitors activate expression of p21Waf1 and 16INK4A in a p53 independent manner. Changes in the expression of these cell cycle regulators by an increase in histone acetylation may be responsible for cell cycle arrest and antitumor activity by HDAC inhibitors. The target molecule of leptomycin B (LMB), a potent antitumor agent, was genetically and biochemically identified as CRM1, a protein reported as being required for chromosome structure control. We showed that CRM1 was a receptor for the nuclear export signal (NES) and that LMB inhibited nuclear export of proteins. Using LMB, we identified a novel NES in fission yeast transcription factor Pap1, the function of which is abolished by oxidative stress in a manner conserved in eukaryotes. PMID- 10667201 TI - The dual role of cytoskeletal anchor proteins in cell adhesion and signal transduction. AB - beta-Catenin and plakoglobin are homologous proteins having a dual role in cell adhesion and in transactivation together with LEF/TCF transcription factors. Overexpression of plakoglobin suppresses tumorigenicity, whereas increased beta catenin levels are considered oncogenic. We compared the nuclear translocation and transactivation by beta-catenin and plakoglobin. Overexpression of each protein resulted in nuclear translocation and formation of structures that also contained LEF-1 and vinculin with beta-catenin, but not with plakoglobin. Transfection of LEF-1 translocated endogenous beta-catenin, but not plakoglobin into the nucleus. Chimeras of the Gal4 DNA-binding domain and the transactivation domains of either plakoglobin or beta-catenin were equally potent in transactivation, but induction of LEF-1-responsive transcription was higher with beta-catenin. Overexpression of wt plakoglobin or mutant beta-catenin lacking the transactivation domain induced nuclear accumulation of the endogenous beta catenin and LEF-1-responsive transactivation. The nuclear localization and constitutive beta-catenin-dependent transactivation in SW480 cancer cells were inhibited by overexpressing cadherin or alpha-catenin. Moreover, transfecting the cytoplasmic tail of cadherin inhibited transactivation, by competition with LEF-1 in the nucleus for beta-catenin binding. The results indicate that (1) plakoglobin and beta-catenin differ in nuclear translocation and complexing with LEF-1 and vinculin, (2) LEF-1-dependent transactivation is mainly driven by beta catenin, (3) cadherin and alpha-catenin can sequester beta-catenin, inhibit its transcriptional activity, and antagonize its oncogenic action. PMID- 10667202 TI - Cytoskeletal tumor suppressors that block oncogenic RAS signaling. AB - Several distinct peptides or drugs that block the Rho family GTPases-mediated pathways were found to suppress RAS-induced malignant phenotype. They include (1) C3 enzyme that selectively inactivates Rho, (2) ACK42, a peptide that blocks the interaction of CDC42 with its effectors such as ACKs, (3) PAK18, a peptide that blocks the activation of PAK and membrane ruffling, and (4) actin-binding drugs, chaetoglobosin K (CK) and MKT-077, that block membrane ruffling by capping and bundling actin filaments, respectively. PMID- 10667203 TI - Antiangiogenic domains shared by thrombospondins and metallospondins, a new family of angiogenic inhibitors. AB - The growth of solid tumors has been shown to depend on neovascularization. By understanding the mechanisms that control the neovascular response, it may be possible to design therapeutic strategies to selectively prevent or halt pathologic vascular growth and restrain cancer progression. Thrombospondin-1 is an extracellular matrix protein that among several functions suppresses capillary growth in angiogenesis assays. We have demonstrated that within the context of the mammary gland TSP1 can modulate normal development of blood vessels. Expression of TSP1 in transgenic animals under the control of the MMTV promoter was associated with a 50-72% reduction in capillary growth. In addition, TSP1 reduced tumor size in transgenic overexpressors. The data suggest an important role for TSP1 in modulating vascular growth in both normal and pathologic tissues. The antiangiogenic region of TSP1 has been mapped to the type I (properdin) repeats. To identify novel proteins with such a domain, we have cloned two cDNAs (METH-1 and METH-2) which also have antiangiogenic properties. In addition to carboxyterminal thrombospondin-like domains they also contain metalloproteinase and disintegrin sequences. Expression of both proteins is broad but nonoverlapping. Recombinant fragments from these sequences have strong antiangiogenic potential in the CAM and cornea pocket assays. At the same molar ratio, METH-1 and METH-2 are about 20-fold more potent than TSP1. We predict that these proteins are likely endogenous modulators of vascular growth with relevant therapeutic potential in cancer and other disease states. PMID- 10667204 TI - Immunotherapy of cancer. AB - The goal of cancer treatment is to develop modalities that specifically target tumor cells, thereby avoiding unnecessary side effects to normal tissue. Vaccine strategies that result in the activation of the immune system specifically against proteins expressed by a cancer have the potential to be effective treatment for this purpose. An early vaccine approach that was developed by our group involves the insertion of the granulocyte-macrophage colony stimulating factor (GM-CSF) gene into cancer cells that are then used to immunize patients. These genetically modified tumor cells produce the immune activating protein GM CSF in the local environment of the tumor cells, specifically activating the patient's T cells to eradicate cancer at metastatic sites. We have performed many studies that demonstrate that this vaccine can cure mice of cancer. We recently demonstrated that this approach can activate an immune response in patients with renal cell carcinoma. We are currently testing a similar approach in patients with pancreatic cancer. Until recently, whole tumor cells were used to produce the vaccine because the proteins expressed by the tumor cells that can be recognized by the immune system were unknown. However, recent advances have allowed the identification of many of the proteins expressed by some cancers. In addition, significant attention has been focused on the mechanisms by which antitumor immunity can be modulated. These active areas of research will undoubtably lead to the development of more specific and more potent vaccine strategies in the near future. The first part of this paper focuses on data from two recent clinical trials that evaluated the whole tumor cell approach. The second part of this paper discusses some of the more exciting antigen-specific vaccine approaches that are under development for the treatment of cancer. PMID- 10667205 TI - Intracellular signaling of the TGF-beta superfamily by Smad proteins. AB - TGF-beta is a potent inhibitor of cell growth, and accumulating evidence suggests that perturbation of the TGF-beta signaling pathway leads to tumorigenesis. Smads are recently identified proteins that mediate intracellular signaling of the TGF beta superfamily. Smads 2 and 3 are phosphorylated by the TGF-beta type I receptor. Smad4 was originally identified as a candidate tumor suppressor gene in pancreatic cancers. Smads 2 and 3 form complexes with Smad4 upon TGF-beta stimulation. The heteromeric Smad complexes translocate into the nucleus, where they activate expression of target genes. Our recent study demonstrated that Smads exist as monomers in the absence of TGF-beta. Smads 2 and 3 form homo- as well as hetero-oligomers with Smad4 upon ligand stimulation. Both homo-oligomers and hetero-oligomers directly bind to DNA, suggesting that the signaling pathway of Smads may be multiplex. Smads 2 and 3 associate with transcriptional coactivators such as p300 in a ligand-dependent manner, p300 enhances transactivation by TGF-beta, suggesting that coactivators link Smads to the basal transcriptional machinery. A missense mutation of Smad2 identified in colorectal and lung cancers was introduced to Smad3. The mutant, Smad3(DE), blocked the activation of wild-type Smad2 and Smad3. Thus, the missense mutation not only disrupts the function of the wild-type Smad but also creates a dominant-negative Smad, which could actively contribute to oncogenesis. PMID- 10667206 TI - Bacterial toxins inhibiting or activating small GTP-binding proteins. AB - Amino acids located on the switch 1 or switch 2 domains of small GTPases of the Ras and Rho family are targets of several bacterial toxins. Exoenzyme C3 from Clostridium botulinum ADP-ribosylates specifically Rho at R43 and prevents the recruitment of Rho on the cell membrane. This blocks the downstream effects of the Rho GTPase. However, exoenzyme C3 is not a toxin, and chimeric proteins fusing C3 with the B moiety of either diphtheria toxin or Pseudomonas aeruginosa exotoxin A have been produced to intoxicate cells with low concentration of C3. C. difficile toxin B modifies by glucosylation Rho on T37 and Rac and Cdc42 on T35. Glucosylation of Rho, Rac, and Cdc42 blocks the binding of these GTPases on their downstream effectors. C. sordellii lethal toxin modifies Ras, Rap, and Rac on T35 by glucosylation. Cytotoxic necrotizing factor 1 (CNF1), from uropathogenic Escherichia coli strains, deamidates Q63 of Rho into E63, thereby blocking the intrinsic or GAP-mediated GTPase of Rho. This allows permanent activation of Rho. Thus, Rho GTPases are targets for three different toxin activities. Molecular mechanisms of these toxins are discussed. PMID- 10667207 TI - Farnesyltransferase inhibitors. Preclinical development. AB - The Ras proteins are low molecular weight GTP binding proteins that function in the regulation of the transduction of growth proliferative signals from the membrane to the nucleus. Oncogenically mutated ras genes are found in approximately 25% of all human cancers. Localization of the Ras oncoproteins to the inner surface of the plasma membrane is essential for their biological activity. This observation suggested that the enzyme that mediates the membrane localization, farnesyl-protein transferase (FPTase), would be a target for the development of novel anticancer agents. We have developed potent, cell-active inhibitors of FPTase that exhibit antiproliferative activity in cell culture and block the morphologic alterations associated with Ras-induced transformation of mammalian cells in monolayer cultures. In vivo, these compounds block the growth of ras-transformed fibroblasts in a nude mouse xenograft model and block the growth and, in some cases, cause regression of mammary and salivary tumors in several strains of ras transgenic mice in the absence of any detectable side effects. The results of our preclinical studies and those of others suggest that FTIs may have utility against a variety of human cancers, a hypothesis that is currently being tested in the clinic. PMID- 10667208 TI - Selective inhibition of ras-transformed cell growth by a novel fatty acid-based chloromethyl ketone designed to target Ras endoprotease. AB - A novel fatty acid-based chloromethyl ketone, UM96001, which was designed to be a Ras C-terminal sequence-specific endoprotease inhibitor, at low micromolar concentrations (1-5.0 microM), potently inhibits ras-transformed rat kidney cell growth, whereas the growth of untransformed normal rat kidney cells is not affected under the same conditions. UM96001 almost completely blocks the anchorage-independent clonogenic growth of ras-transformed rat and human cancer cells at low micromolar concentrations. Inhibition of ras-transformed rat and human cancer cell growth by UM96001 may occur via the mechanism of selective induction of apoptosis of the cells. Furthermore, TPCK and BFCCMK, the known selective inhibitors of Ras C-terminal sequence-specific endoprotease, also yield similar inhibition results. These results provide the first experimental evidence that the endoproteolysis of Ras oncoproteins may be important for Ras-mediated cell growth and apoptosis. Therefore, the Ras C-terminal sequence-specific endoprotease may be a potential anticancer target. PMID- 10667209 TI - Azatyrosine. Mechanism of action for conversion of transformed phenotype to normal. AB - Azatyrosine [L-beta-(5-hydroxy-2-pyridyl)-alanine] has the unique property of converting ras- or c-erbB-2 transformed phenotype to normal. The administration of azatyrosine also inhibits tumor formation in transgenic mice harboring the normal human c-Ha-ras which is mutated during treatment with various chemical carcinogens. To elucidate the molecular mechanism, we investigated how azatyrosine functions and what are its major targets. Azatyrosine functions downstream of ras; azatyrosine does not alter either the level of GTP-bound Ras or the total amount of Ras. Instead, azatyrosine inhibits the activation of c-Raf 1 kinase by oncogenic c-ErbB-2, resulting in inactivation of AP1. It is interesting that azatyrosine also restores the expression of the rhoB gene, the product of which regulates the formation of actin stress fibers. Azatyrosine is incorporated into cellular proteins to replace tyrosine. Several experiments indicate that replacement of tyrosine is likely to be a cause for its conversion of transformed phenotype to normal. To prove this hypothesis, we are attempting to develop a mutant of tyrosyl-tRNA synthetase that, unlike wild type, can aminoacylate azatyrosine more efficiently than can tyrosine. PMID- 10667210 TI - SCH 51344, an inhibitor of RAS/RAC-mediated cell morphology pathway. AB - RAS interacts with multiple targets in the cell and controls at least two signaling pathways, one regulating extracellular signal-regulated kinase (ERK) activation and the other controlling membrane ruffling formation. These two pathways appear to act synergistically to cause transformation. Human smooth muscle alpha-actin promoter is repressed in RAS-transformed cells and derepressed in revertant cell lines, suggesting that it is a sensitive marker to follow phenotypic changes in fibroblast cells. SCH 51344 is a pyrazoloquinoline derivative identified on the basis of its ability to derepress alpha-actin promoter in RAS-transformed cells. Previous studies have shown that SCH 51344 is a potent inhibitor of RAS transformation. However, SCH 51344 had very little effect on the activities of proteins in the ERK pathway, suggesting that it inhibits RAS transformation by a novel mechanism. Recently, we have demonstrated that SCH 51344 specifically blocks membrane ruffling induced by activated forms of H-RAS, K-RAS, N-RAS, and RAC. Treatment of fibroblast cells with this compound had very little effect on RAS-mediated activation of ERK and JUN kinase activities. SCH 51344 was effective in inhibiting the anchorage-independent growth of Rat-2 fibroblast cells transformed by the three forms of oncogenic RAS and RAC V12. These results indicate that SCH 51344 inhibits a critical component of the membrane ruffling pathway downstream from RAC and suggest that targeting this pathway may be an effective approach to inhibiting transformation by RAS and other oncogenes. PMID- 10667211 TI - Activation of apoptosis and its inhibition. AB - The induction of apoptosis, or controlled cell death, by various stimuli has been shown to activate a cascade of endoproteases, called caspases, that cleave numerous cellular proteins necessary for cellular homeostasis. This review discusses this family of proteases together with a variety of mammalian and viral regulatory proteins that act to control this activation. PMID- 10667212 TI - Strategies to adapt adenoviral vectors for targeted delivery. AB - The utility of current generation adenoviral vectors for targeted, cell-specific gene delivery is limited by the promiscuous tropism of the parent virus. To address this issue, we have developed both genetic and immunologic methods to alter viral tropism. Immunologic retargeting has been achieved via conjugates comprised of an antifiber knob Fab and a targeting moiety consisting of a ligand or antireceptor antibody. Gene delivery by this approach has been accomplished via a variety of cellular pathways including receptors for folate, FGF, and EGF. In addition to cell-specific gene delivery, this strategy has allowed enhanced gene delivery to target cells lacking the native adenoviral receptor, CAR. Of note, this specific and extended gene delivery allowed enhanced survival in murine models of human carcinoma via cancer gene therapy. Genetic strategies to alter adenoviral tropism have included both fiber modification and fiber replacement. In the former, we have identified the HI loop of fiber as a propitious locale for introduction of heterologous peptides. Incorporation of an RGDC peptide at this locale allowed gene delivery via cellular integrins with dramatic efficiency augmentations. As a strategy to achieve both new tropism as well as to ablate native tropism, methods have been developed to replace the fiber protein with heterologous motif which preserves the key trimeric quaternary structure of fiber and allows for propagation. Such a fiber-replacement virus has been rescued and has demonstrated capacities consistent with its utility as a novel vector agent. These strategies have allowed the achievement of cell specific gene delivery via adenoviral vectors and thus have the potential to enhance the utility of this vector agent. PMID- 10667213 TI - Homeodomain-derived peptides. In and out of the cells. AB - The internalization of homeodomains and of homeopeptides derived from the third helix of the homeodomain of Antennapedia, a Drosophila transcription factor, is used by some investigators to target exogenous hydrophilic compounds into live cells. In addition to this very practical aspect of drug delivery, translocation across biologic membranes of peptides subsequently addressed to the cell cytoplasm and nucleus raises several questions. A first series of questions pertains to the mechanism of translocation. Thanks to the synthesis of several peptides derived from the third helix of the Antennapedia homeodomain, we began to investigate the mechanism of translocation and we have shown that it is not dependent upon the presence of a chiral receptor and probably involves the formation of inverted micelles. A second series of questions is related to the physiologic significance of the phenomenon. In a first approach, we demonstrated that some full-length homeoproteins are internalized and secreted in vitro. The mechanism of internalization is probably similar to that of the homeodomain or of its third helix, but secretion involves a different mechanism which requires an association with specialized intracellular membranous structures. The existence of specific mechanisms for homeoprotein internalization and secretion suggests that this class of transcription factors may have important signaling properties. PMID- 10667214 TI - In vitro evaluation of a novel 2,6,9-trisubstituted purine acting as a cyclin dependent kinase inhibitor. PMID- 10667215 TI - Homologous recombination between heterologs during repair of a double-strand break. Suppression of translocations in normal cells. PMID- 10667216 TI - Selection of genetic suppressor elements conferring resistance to DNA topoisomerase II inhibitors. PMID- 10667217 TI - BAP1, a candidate tumor suppressor protein that interacts with BRCA1. PMID- 10667218 TI - Histone deacetylase inhibitor activates the p21/WAF1/Cip1 gene promoter through the Sp1 sites. AB - Trichostatin A (TSA), a specific histone deacetylase inhibitor, induces histone hyperacetylation and modulates the expression of some genes. We examined the effects of TSA on MG63 cells. TSA induced growth arrest and expression of the p21/WAF1/Cip1 protein. A close correlation between the level of histone acetylation and induction of the p21/WAF1/Cip1 protein was detected. Using several mutant p21/WAF1/Cip1 promoter fragments, mutation of either of two Sp1 sites at -82 or -69 of the p21/WAF1/Cip1 promoter reduced the responsiveness to TSA. This finding indicates that TSA activates the p21/WAF1/Cip1 promoter through the Sp1 sites in a p53-independent manner. PMID- 10667219 TI - Selective induction of cyclin-dependent kinase inhibitors and their roles in cell cycle arrest caused by trichostatin A, an inhibitor of histone deacetylase. PMID- 10667220 TI - Identification of a novel nuclear export signal sensitive to oxidative stress in yeast AP-1-like transcription factor. PMID- 10667221 TI - Melanoma cell lines contain a proteasome-sensitive, nuclear cytoskeleton associated pool of beta-catenin. PMID- 10667222 TI - Truncated form of beta-catenin and reduced expression of wild-type catenins feature HepG2 human liver cancer cells. PMID- 10667223 TI - Induction of apoptosis by gelsolin truncates. PMID- 10667224 TI - Reexpression of the major PKC substrate, SSeCKS, correlates with the tumor suppressive effects of SCH51344 on rat-6/src and rat-6/ras fibroblasts but not on rat-6/raf fibroblasts. PMID- 10667225 TI - Subtractive cDNA cloning and characterization of genes induced by all-trans retinoic acid. PMID- 10667226 TI - Purification and functional characterization of a novel protein encoded by a retinoic acid-induced gene, RA28. PMID- 10667227 TI - Augmented inhibition of tumor cell proliferation in combined use of electroporation with a plant toxin, saporin. PMID- 10667228 TI - Expression of matrix metalloproteinases and their inhibitors in human brain tumors. AB - Sixty human brain tumors, including grade I meningiomas, schwannomas, and pilocytic astrocytomas, grade II astrocytomas, grade III anaplastic astrocytomas and oligodendrogliomas, and grade IV glioblastomas and lung and melanoma metastases were analyzed for expression of four matrix metalloproteinases (MMPs), two tissue inhibitors of MMPs (TIMPs), and MMP activity. No marked correlation was found between MMP expression and the degree of malignancy. Western blotting analysis revealed a more uniform pattern of distribution of MMP-2 (gelatinase A) than of MMP-9 (gelatinase B) and MMP-12 (metalloelastase) among tumors. All 60 tumors showed a similar pattern of activity in zymography, MMP-2 being the major species detected. Interestingly, TIMP-1 and TIMP-2 expression levels were low in tumors of grade III but significantly higher in tumors of grade I, particularly schwannomas. Altogether, these data suggest that: (1) the balance between MMP-2 and TIMP-2 is important in human brain tumors; and (2) TIMP expression may be a valuable marker for tumor malignancy. PMID- 10667229 TI - Plasmin-depletion therapy. A new approach to overcoming tumor cell drug resistance. PMID- 10667230 TI - Polysulfated heparinoids selectively inactivate heparin-binding angiogenesis factors. AB - Angiogenesis is a prerequisite for tumor expansion and metastasis. The angiogenic potential of the heparin-binding growth factors acidic fibroblast growth factor (FGF) and basic FGF has been demonstrated in various publications. We studied the inhibitory effects of suramin and the polysulfated heparinoids pentosan polysulfate, dextran sulfate, and fucoidan on the action of FGF. As an experimental model, we used the adrenal cancer cell line SW 13, whose anchorage independent growth depends on the presence of FGF. The polysulfated heparinoids inhibited FGF-induced growth and binding to the receptor at an IC50 of 0.5-3 micrograms/ml. Suramin inhibited FGF at an IC50 of 100 micrograms/ml. The polysulfated heparinoids exerted no effect on IGF-1 or TGF alpha-related growth. Suramin inhibited the anchorage-independent growth induced by IGF-1 or TGF alpha only at an IC50 of 100 micrograms/ml. Our results indicate that suramin inhibits growth factors in a nonselective way. By contrast, polysulfated heparinoids exert a selective inhibitory effect on heparin binding angiogenesis factors at an IC50, which is 100 times below the IC50 of suramin. Therefore, the administration of polysulfated heparinoids might become a novel approach to tumor therapy based on blocking angiogenesis. PMID- 10667231 TI - RPR 130401, a nonpeptidomimetic farnesyltransferase inhibitor with in vivo activity. PMID- 10667232 TI - Cellular effects of a new farnesyltransferase inhibitor, RPR-115135, in a human isogenic colon cancer cell line model system HCT-116. PMID- 10667233 TI - The farnesyltransferase inhibitor L-744,832 inhibits the growth of astrocytomas through a combination of antiproliferative, antiangiogenic, and proapoptotic activities. PMID- 10667234 TI - Isolation of farnesyltransferase inhibitors from herbal medicines. PMID- 10667235 TI - Farnesyltransferase inhibitor-induced regression of mammary tumors in TGF alpha and TGF alpha/neu transgenic mice correlates with inhibition of map kinase and p70s6 kinase phosphorylation. PMID- 10667236 TI - Inhibition of protein tyrosine kinase activity by 1a-docosahexaenoyl mitomycin C. PMID- 10667237 TI - Activation of a 36-kD MBP kinase, an active proteolytic fragment of MST/Krs proteins, during anticancer drug-induced apoptosis. PMID- 10667238 TI - Peptides mimicking sialyl-Lewis A isolated from a random peptide library and peptide array. PMID- 10667239 TI - B4112, a novel tetramethylpiperidine-substituted phenazine that inhibits the proliferation of multidrug-resistant cancer cell lines. PMID- 10667241 TI - Construction of a cell-permeable CDC42 binding fragment of ACK that inhibits v-Ha Ras transformation. PMID- 10667240 TI - The anti-RAS cancer drug MKT-077 is an F-actin cross-linker. PMID- 10667242 TI - Rational development of cell-penetrating high affinity SH3 domain binding peptides that selectively disrupt the signal transduction of Crk family adapters. Amgen Peptide Technology Group. PMID- 10667243 TI - Anti-HER2 immunoliposomes for targeted drug delivery. PMID- 10667245 TI - Management of head and neck tumors: what are the challenges for the third millennium? Abstracts. PMID- 10667247 TI - Cumulative author and subject index for volumes 1-13. PMID- 10667244 TI - Recombinant cytotoxins specific for cancer cells. PMID- 10667249 TI - The Physiologic Society proceedings of the scientific meeting held at University of Glasgow, 14-16 September 1999. Abstracts. PMID- 10667246 TI - 10th International Dresden Symposium on Lipoproteins and Atherosclerosis. Dresden, Germany, 9-11 December 1999. Abstracts. PMID- 10667251 TI - Cumulative 22-year index, volumes 1-22, 1978-1999. PMID- 10667250 TI - Ten-year cumulative index, volumes 1-10, 1990-1999. PMID- 10667252 TI - International Society of Psychoneuroendocrinology (ISPNE) XXXth Congress. Orlando, Florida, USA. 30 July-3 August 1999. Abstracts. PMID- 10667248 TI - International Society for Heart and Lung Transplantation 20th annual meeting and scientific sessions. Osaka, Japan, April 5-8, 2000. Abstracts. PMID- 10667253 TI - ESTRO-EORTC meeting on Radiation for Benign Disease: Current Status and Possible Perspectives. Brussels, Belgium, 10-13 October 1999. Abstracts. PMID- 10667254 TI - [Consensus on the management of obesity, Switzerland 1999]. PMID- 10667256 TI - Bibliography of toxinology. PMID- 10667255 TI - V Symposium of the Brazilian Society on Toxinology. Understanding and exploiting toxins for the XXI century. Angra dos Reis, Rio de Janeiro, Brazil. 13-18 September 1998. Abstracts. PMID- 10667257 TI - Master index to volumes 71-80. PMID- 10667258 TI - [Anthropometric characteristics of obese women before and after weight reduction]. AB - Anthropometric changes in 53 premenopausal obese women, 25-45 year-old, after an obesity treatment, were analyzed. Before and immediately after treatment, midarm, abdomen, waist, hip, midthigh and midcalf circumferences, as well as tricipital, bicipital, suprailiac, subscapular, abdominal and calf skinfold thickness were measured. The later averaged over percentil 75th. All final measurements, except midcalf circumference, abdomen/waist and hip/midthigh indexes, were significantly lowers as compared with the initial values. Body weight decreased on average 8.9 kg, and skinfold thickness mean fell below the 75th percentile. Percentage body fat decreased 18.3%, and body mass index 11%. Initial principal component analysis results, exhibited three chief characteristics of obesity: general adiposity; trunk vs. extremity fatness, and upper vs. lower fatness. Body fat distribution pattern and body composition were modified as a result of the treatment. Therefore, the morbidity risk associated with them, was substantially reduced. PMID- 10667259 TI - [Maternal height and growth of Chilean premature infants]. AB - The aim of this study was to analyze the association between maternal age and growth of infants born preterm through the first 4 mo. of age. We prospectively studied 80 infant born at the hospital Herminda Martin in Chilean between January and September 1995 with birthweight < 2500 g and gestational age < or = 36 weeks; those born small for date, with congenital malformations or developing chronic diseases were excluded. The modified Graffar was applied to study socioeconomic status and growth, morbidity and type of feeding was monthly registered at Primary Care Centers where they were controlled. The maternal height was categorized as small (< -1 SD, n = 14), normal (> -1 SD and < +1 SD, 147.6-161.8 cm, n = 52) or tall (> +1 SD, n = 14). Sons from tall mothers presented better W/A z score at 4 mo than those from normal or small mothers (0.85 +/- 0.8 vs 0.31 +/- 0.6, p < 0.03 and 0.85 +/- 0.8 vs 0.15 +/- 0.8, p < 0.04). Length gain through the 4 mo was also better of infants with tall mothers (15.3 +/- 1.4 vs 13.7 +/- 2.3 and 13.6 +/- 2.2 cm, ANOVA p < 0.04), reaching better z-scores (0.2 +/- 0.3 vs -0.7 +/- 0.6 and -0.9 +/- 0.9, ANOVA p < 0.0001). The maternal schooling > 8 y was also associated to infant growth: those with tall mothers presented better L/A z-score at 4 mo than those with normal or small mothers (0.27 +/- 0.3 vs -0.89 +/- 0.7 and -0.85 +/- 0.5 p < 0.001). Exclusive breast milk was present in 7% of tall, 25% of normal and 0% of small mothers. No differences in morbidity were observed between groups. We conclude that Chilean infants born preterm from mothers > 1.61 m present a better growth that those with smaller mothers since the first 4 months of age. PMID- 10667260 TI - [Nutritional study relative to proteins, energy, and calcium in children eating school meals]. AB - Food consumption and nutritional status were evaluated on 419 children having meals at school located in the surrounding of Santa Fe (Argentina). The protein, energy and calcium content of the meals were analyzed. Anthropometrical evaluation was made by the following indicators: weight to age, height to age and weight to height. Biochemical evaluation to a sample of the children, was made by urea/creatinine and calcium/creatinine indicators. Recommended protein consumption is achieved but a biological efficiency loss is detected, probably due to an insufficient energy intake. School meals provide about 50% of the daily energy requirements. Regarding calcium consumption, the dinner cover about 15% of the recommendations but they achieve 47-49% with the cup of milk. As in the case of energy, the calcium daily intake is insufficient as we can see by the calcium/creatinine indicator which shows that a 60% of the people are below of the normal values. All these results suggest that is necessary to increase the energy intake and calcium consumption to improve the nutritional status. PMID- 10667261 TI - [Effect of breast feeding and psychosocial variables upon psychomotor development of 12-month-old infants]. AB - This study evaluates the participation of psychosocial variables in the relation between breast feeding (BF) and psychomotor development (PMD) in dyads with different BF duration. We assessed 138 mother-infant dyads, divided in two groups: 86 received BF as unique source of milk feeding for at least 6 months (prolonged BF group) and 52 were weaned before 45 days of age (early weaning group). General information about pregnancy, delivery and feeding was collected in a non experimental prospective design. At 6-7 months of age a milk feeding situation was observed at home, and mother-infant interactional patterns were recorded through a specially designed scale. At 12 months of age the PMD was assessed (Bayley Scales of Infant Development). Infant temperament, home stimulation, mother depression and family stress were also measured. Similar family characteristics were observed in both study groups. Early bonding and first feeding experiences were different, both reported as better in the prolonged BF group. Moreover, dyads of this group showed a higher variety and quality of mother-infant interactional patterns during feeding, with a higher synchrony and reciprocity in the relationship. Mean Mental Development Index (MDI) and Psychomotor Developmental Index (PDI) were similar in both groups. Explicatory variables for MDI and PDI are different in both study groups. Dyads who attained prolonged BF conform from a psychosocial perspective--a different group than the early weaned. PMID- 10667262 TI - [Basal metabolic rate is overestimated by predictive equation in college-age women of Rio de Janeiro, Brazil]. AB - Since the World Health Organization suggested predictive equations for basal metabolic rate (BMR) in 1985 there has been great interest in their validity in different populations worldwide. It has been shown that these equations overestimate BMR in some populations, particularly the ones living in the tropics. There is limited new information on BMR in segments of the Brazilian population. Therefore, the aim of the present study was to compare measured with estimated BMR using some published predictive equations in 50 college students from Rio de Janeiro, Brasil. BMR was measured by indirect calorimetry and the predictive equations used were the ones published by: FAO/WHO/UNU (1985); Harris & Benedict (1919), and Henry & Rees (1991). Estimated BMRs were significantly greater than measured BMR (p < 0.05). Overestimation was greatest with the equation published by Harris & Benedict (18.9%) followed by the ones by FAO/WHO/UNU (12.5%) and Henry & Rees (7.2%). Body composition did not correlate with the overestimation of BMR. More data are necessary so that appropriate predictive equations can be developed for the Brazilian population. PMID- 10667263 TI - [Erythrocyte protoporphyrin during recovery from malnutrition in rats]. AB - Interrelationships between Erythrocyte Protoporphyrin (EP), dietary Iron/Protein ratio (Fe/Prot) and Fe liver content (Feh) were studied during nutritional recovery in an experimental model: weanling female Wistar rats (To) were depleted with a protein-free diet (LP), losing 20% of their initial body weight. Then they were recovered until 45 days of age (T45) with diets containing: casein: 20 g/100 g; Fe (ammonium Fe citrate) (ppm.): 0, 75 or 100 (groups A1, A2 and A3, respectively). Hematocrit, Hemoglobin (Hb) (g/dL). Erythrocyte Protoporphyrin (EP) (microgram/dL Red Blood Cells) and Feh (microgram) were determined at To, LP and T45. Results were compared with control rats (C) fed with 20% of casein and Fe, 50 ppm. EP: a) decreased in C from To to T45 (99 +/- 24; 36 +/- 9; p < 0.01); b) increased in A1 and A2 at T45 (123 +/- 21; 93 +/- 29, respectively, p < 0.01) while A3 did not show significant difference (45 +/- 7) regarding to C: c) correlated inversely with Feh. According to the inverse correlation between EP and Fe/Prot (r = -0.99), we found that 92 ppm was an adequate Fe amount to prevent EP increase. These results confirm that during recovery from undernutrition EP depends on iron liver content, being an adequate indicator of iron nutritional status; therefore, EP would be useful as a predictor of the optimum Fe/Prot ratio for nutritional recovery. PMID- 10667264 TI - [Rural development, household food safety, and nutrition in western Honduras]. AB - The authors studied the impact of a rural development project on household food security and nutrition. A quasi-experimental study design was used to compare the experience of members of thirteen Honduran small-holder farmers groups which had already received a year of credit and technical assistance, with another thirteen groups which had just joined the project, and thirteen control communities. All these communities were followed-up for one year (March/April 1997-March/April 1998). Farmers participating in the project showed a greater increase in maize stores than farmers in the control communities (p = 0.01), but did not increase their dietary energy consumption. There was, however, a small improvement in their dietary diversity (p = 0.01). The impact of the project on the nutritional status of under 5's was complex. The study underlined the importance of monitoring the impact of programs which may affect food and nutrition. PMID- 10667265 TI - [Evaluation of cookies enriched with corn germ and soy fiber]. AB - The objective of this study was to evaluate four cookie formulations which wheat flour was partially substituted by free-fat corn germ flour and/or soy fiber. Baking quality, protein, fat, ash, dietary fiber, hardness, color, Protein Efficiency Ratio PER and Apparent Digestibility in vivo were determined. A trained panel evaluated color, hardness and fracturability of cookies. Dietary fiber of cookies varied from 8.2 to 24.9% and protein from 11.3 to 12.7%. The source and amount of dietary fiber modified physical, sensory, and nutritional properties of cookies. Cookies formulated with 20% corn germ flour gave the highest PER, Digestibility Aparente in vivo, and acceptance by consumers. This study demonstrated the potential use of free-fat corn germ and soy fiber as functional ingredients. PMID- 10667266 TI - Nutritional evaluation of table bread fortified with defatted soybean and sesame meals. AB - The nutritional value of table bread fortified with 8% defatted soybean meal (DSBM), 12% DSBM and a mixture of 8% DSBM/4% defatted sesame meal (DSM) was assessed with in vivo and in vitro tests. Fortification with DSBM and DSM decreased protein digestibilities (P < 0.05) but improved essential amino acid scores (EAA) and overall nutritional value of the breads. Fortified breads contained twice as much lysine, and consequently a better protein efficiency ratio (PER) than the control bread. The PER of the 8% DSBM/4% DSM bread was similar (P > 0.05) to the 12% DSBM bread. The 8% DSBM fortified bread showed lower PER, amino acid and protein contents than breads fortified with 12% DSBM. In vitro procedures utilized to predict protein digestibilities and PER's provided a close estimation of in vivo results obtained from growing rats. PMID- 10667267 TI - [Sensory characterization of pate from waste salmon (Salmo salar L.) products utilizing response surface methodology]. AB - The goal of the present study was the development of a salmon pate formulation with excellent nutritional and sensory properties, using by product components from de salmon industry. The optimized formulation was obtained using the response surface methodology. A 2n factorial and a composite central rotatable design was applied in the experience. Salt and xanthan hydrocolloid gum were the selected independent variables and sensory quality the response variable. Statistical analysis was utilized to estimate the fitted model. The optimum combination of selected experimental variables were 1.5% salt and 1.35% xanthan gum. Data from the chemical characterization of salmon pate showed an important protein content (20%) greater than liver pate (11.5%) and trout pate (14.7%). With regard to fat content, the concentration of this component in salmon pate was half the concentration found in liver pate. The shelf life of the optimized formula determined at 3 degrees C and 18 degrees C were 18 days and 8 days respectively. PMID- 10667268 TI - Antioxidant concentration effect on stability of Brazil nut (Bertholletia excelsa) crude oil. AB - Shelled and broken Brazil nuts easily lose quality, if not properly stored. Pressing is an alternative use of these nuts and the crude oil stability was studied. Our previous studies demonstrated that TBHQ (200 mg kg-1) was very efficient to prevent rancidity development in oils bottled in brown and clear glass. As TBHQ has higher price than other phenolic antioxidants like BHT and BHA, an oven test (at 63 degrees C) was conducted to determine the economical and best concentration of TBHQ for Brazil nut crude oil. An assay at ambient temperature was conducted in brown and clear glass flasks with and without the economical concentration of TBHQ calculated (83 mg kg-1) for 90 days. Acid, peroxide, and iodine indices and the absorptivity at 232 nm were determined. TBHQ, even at this low dosage, was very efficient in both brown and clear glass flasks. Peroxide value increased from 11.5 meq O2 kg-1 to average 15 and 35, in TBHQ and control samples after 90 days. The absorptivity at 232 nm remained at 1.3 in samples with TBHQ while the control increased to 1.6. PMID- 10667269 TI - [Presence of Clostridium perfringens in meat-based preparations in public food services in central San Jose, Costa Rica]. AB - In Costa Rica there are a large number of public food services distributed along the country, where a considerable number of people eat daily. Clostridium perfringens is a bacteria associated with foodborne illness related, especially, to meat products kept for long time at temperatures under 70 degrees C. The aim of this study was to evaluate the public food services that use water baths for keeping food hot in order to establish the presence of C. perfringens in cooked bovine meat dishes and to evaluate the enterotoxigenic capacity of the strains isolated. 81 samples of cooked bovine meat plates coming from 27 public food services, located in the Central County of San Jose were analyzed. The methodology described by Labbe & Harmon for the isolation of C. perfringens was used in 10 g of sample. Also, the enterotoxigenic capacity of the strains was evaluated using the passive-reverse-latex-agglutination assay from Oxoid. From the 27 public food services analyzed, eight (30%) were positive in the three samplings done, nine (33%) were positive in one or two occasions, and ten (37%) were negative all times. This implies that in 17 (63%) of the establishments studied, the bacteria was isolated at least once. From the 81 preparations studied, 37 (46%) were positive for the bacteria. The temperatures at which food was kept varied from 56 to 82 degrees C, with an average of 68.7 degrees C. From the 37 strains identified as C. perfringens, 12 (32%) were positive for enterotoxin. In conclusion, the presence of C. perfringens in bovine meat dishes, maintained in water baths, represents an important risk for public health, and the temperature at which the preparation is kept is critical for the multiplication of the bacteria. PMID- 10667270 TI - [Determination of clostridium perfringens in pork sausages from the Metropolitan area of Costa Rica]. AB - The presence of C. perfringens was analyzed in 75 samples of pork sausages (chorizo, salchichon and bologna), obtained from five processing plants located in the Metropolitan Area of Costa Rica. Previously and after the biochemical identification of the strains, the most probable number (MPN) of C. perfringens per gram of food was determined and it varied from less than 3 to more than 2.4 x 10(5). There were significant statistical differences (p < 0.005) that support the need of employing biochemical tests for confirming C. perfringens in a given food. C. perfringens was present in 92% of the chorizos, in 28% of the bolognas and in 12% of the salchichon. Every positive sample was tested looking for at least one enterotoxigenic strain, using the reverse passive agglutination latex test; 8% of the tested strains were enterotoxigenic and corresponded to chorizo and bologna from one processing plant and chorizo from another plant. The results obtained in this study show that pork sausages, and not just not processed meats, are important as risk factors for food intoxication by C. perfringens. PMID- 10667271 TI - Physical-chemical composition of in natura goat milk from cross Saanen throughout lactation period. AB - The analyzed milk samples were collected from cross Saanen goats of different ages and different cross breeding types, throughout the lactation period, from September 1996 to December 1997. For the physical-chemical characteristics measured in this experiment, the following average values were obtained, followed by their respective standard deviations: pH (6.69 +/- 0.20); acidity (12.96 +/- 3.64 degrees D); density (1.030 +/- 0.009 mg.cm-3); fat (3.83 +/- 1.04%); crude protein (3.34 +/- 0.73%) and total solids (12.25 +/- 1.94 g.100 g-1). The lactation period influenced the values of acidity, fat, crude protein and total solids; these values decreased during the initial months and increased at the end of the lactation. The correlations were analyzed among the studied characteristics during the lactation, resulting in positive (p < 0.05) acidity/density correlation (r = 0.2115), stand out also the positive correlations (p < 0.01) among fat/total solids (r = 0.7715) and crude protein/total solids (r = 0.6228). PMID- 10667272 TI - [Iron, zinc and copper content of foods commonly consumed in Mexico]. AB - Nutrient composition in foods is necessary for the determination of nutrient intake. Food composition tables used for dietary studies in Mexico do not have information of zinc, iron and copper; when present the values have been extrapolated from laboratory analysis carried out with foods in other countries and regions of the world. In this study zinc, iron, and copper content of 104 plant foods and 32 animal foods was determined. The procedure used was atomic absorption spectrophotometry for the minerals. Foods were grouped into cereals, vegetables, fruits, legumes and animal foods. Zinc content ranged from 0.018 mg/100 g for strawberry to 9.193 mg/100 g for beef. Iron content ranged from 0.113 mg/100 g for yogurt to 19.82 mg/100 g for a commercial cereal which had minerals added during processing. In some foods copper was not found and the highest content was 3.371 mg/100 g in beef liver. This study has provided information on zinc, iron and copper content of the most commonly consumed foods in Mexico. PMID- 10667273 TI - Lipids and fatty acids in roasted chickens. AB - Total lipids from meat portions of breast, thigh, wing, side and back with and without skin from 10 roasted chickens were extracted with chloroform and methanol and gravimetrically determined, and their fatty acids were analysed as methyl esters by gaseous chromatography, using a flame ionization detector and capillary column. The main fatty acids found were: C16:0, C18:1 omega 9, and C18:2 omega 6. The average ratio observed between PUFA/SFA was of 0.98, mainly due to the great concentration of the C18:2 omega 6 fatty acid, with an average of 26.75%. Regarding to the lipids content, the skinless breast showed the lowest content, 0.78 g/100 g, while the back with skin was the one with the highest content, 12.13 g/100 g except for the pure skin, with 26.54 grams of lipids by 100 grams. PMID- 10667274 TI - Immunological alterations in patients with primary tumors in central nervous system. AB - Natural killer (NK) cells play an important role in immune surveillance against tumors. The present work aimed to study the cytotoxic activity of NK cells and T cell subsets in peripheral blood of 13 patients with primary tumors in central nervous system (CNS). As controls 29 healthy subjects with the age range equivalent to the patients were studied. The methods employed were: a) determination of cytotoxic activity of NK cells towards K562 target cells, evaluated by single cell-assay; b) enumeration of CD3+ lymphocytes and their CD4+ and CD8+ subsets defined by monoclonal antibodies; c) the identification of tumors were done by histologic and immunochemistry studies. The results indicated that adults and children with tumor in CNS display reduced percentage of total T cells, helper/inducer subset and low helper/suppressor ratio. The cytotoxic activity of NK cells was decreased in patients with CNS tumors due mainly to a decrease in the proportion of target-binding lymphocytes. These results suggest that cytotoxic activity of NK cells may be affected by the immunoregulatory disturbances observed in patients with primary tumors in CNS. PMID- 10667275 TI - The Kohs' blocks test as an important instrument to investigate the visuo-spatial impairments in myotonic dystrophy. Part I. Quantitative and qualitative analysis. AB - This study presents the performance of 39 cases of myotonic dystrophy on Kohs' blocks test (21 females and 18 males, age range from 9 to 70 years). On this test, the patients have to reproduce figures from models previously showed to them. Some of the patients had some kind of professional activity, while others had never exerted a professional occupation. The patients denoted considerable difficulty to perform the test. Some cases constructed entirely different figures in comparison to the presented drawings, translating visuo-spatial and constructional disabilities. The performance was insufficient in 71.4% of the cases. These cases solved less than 50% of the test. The levels of analysis and synthesis were severely impaired. A total of 18 cases got less than 10 points, not reaching 20% of the test. The results showed the sensitivity of this test in detecting visuo-spatial impairment in myotonic dystrophy. PMID- 10667276 TI - Sleep characteristics in children in the isolated rural African-Brazilian descendant community of Furnas do Dionisio, State of Mato Grosso do Sul, Brazil. AB - Developmental and cultural factors affect sleep habits in childhood. The objective of this research was to determine sleep habits of children in the isolated rural African-Brazilian community of Furnas do Dionisio. Mato Grosso do Sul, Brazil. The members of this community are closely related descendants of the ex-slave Dionisio, and remained in relative geographical isolation for about a century. Sleep characteristics of 55 children (35 M; 20 F), 2 to 10 year olds, were evaluated in interviews with their mothers. The results showed that cosleeping, in the same bed with family members, was present in 80.0% of the 2-3 year olds; decreasing to 25.0% of the 8-10 year olds. Only 5.4% of the children slept alone in their own bedroom. Mean number of persons per bedroom was 2.8. Only 7.0% of the bedrooms had TV; 98.1% slept in silence. The data obtained support the need to weigh cultural factors influence on sleep. PMID- 10667277 TI - A case-control study of a benign electroencephalographic variant pattern. AB - Wicket spikes (WS) are a benign electroencephalogram (EEG) variant, seen mainly in adults, during somnolence, in the temporal regions, in many clinical situations. WS can appear in trains or isolatedly, sometimes being difficult to differentiate from epileptiform activity. We reviewed 2,000 EEG's, found 65 with WS (3.25%) and compared them with 65 normal EEG without WS. There was statistically significant (SS) association between WS and age over 33; adolescent age was correlated to absence of WS and age over 65, to the presence of WS; there was an inverse correlation between WS and epilepsy, related to differences in age; a SS association with cerebrovascular disorders disappeared after controlling for age; a SS correlation with headache was also related to age; female predominance was not SS. There was a great variety of clinical situation associated with WS. We conclude that WS are a inespecific normal variant of the EEG that is age-related. PMID- 10667278 TI - Continuous spike-waves during slow waves sleep. A clinical and electroencephalographic study in fifteen children. AB - We report on the clinical and EEG features of 15 patients with the syndrome of "continuous spike waves during slow wave sleep" (CSWSS). The differential diagnosis of CSWSS includes benign epilepsy of childhood with centro-temporal spikes, and Landau-Kleffner and Lennox-Gastaut syndromes. We found normal CT and MRI features in 6 cases, periventricular leukomalacia with and without diffuse brain atrophy in 4 cases and hydrocephalus in 1 case. There was no association between specific neurological findings and CSWSS. Nine of our cases had relatively focal discharges, like some cases from the literature. The occurrence of CSWSS appears to be age-related, generally between the ages of 5 to 12 years, with a strong temporal relation to the neuropsychological deterioration in its nature, severity and prognosis. We believe that this striking disorder has been overlooked and that routine sleep EEG studies on epileptic children may disclose additional cases of CSWSS. PMID- 10667279 TI - [Video-polygraphic-EEG study in the full-term newborn with low birthweight for their gestational age]. AB - Video-polygraphic-EEG studies were performed in the first 24 life-hours of 26 healthy full-term newborns without perinatal injuries. The neurological examination and cranial ultrasonography were normal. The newborns were divided into two groups: one, with full-term appropriate--birth weight 11 newborns (control group) and the other with full-term low-birth weight 15 newborns. Thirteen newborns of the second group had video-polygraphic-EEG study abnormalities. The most frequent abnormalities were found in 11 cases, as far as sleep architecture is concerned. Also, when compared with the control group, 8 cases of an excessive amount of startles and 2 cases of low behavior activities were found. The results demonstrate the usefulness of video-polygraphic-EEG study in the full-term newborns with intra-uterine growth retard. This examination was sensitive to detect behavior, sleep architecture and EEG standard differences in the low birth-weight newborns as to the control group. PMID- 10667280 TI - [Electroencephalographic modification in Down syndrome]. AB - The frequency of epilepsy in Down syndrome (DS) has been reported in literature varying from 6 to 17%. A typical electroencephalographic (EEG) pattern has not been established for this condition. There is a great variation on EEG abnormalities and most of them are not associated to behavior alterations or neurological signs. The aim of this study was to establish epidemiological and electroencephalographic parameters in institutionalized patients with clinical diagnosis of DS. We studied 77 individuals of both sexes, age ranging from 0-38 years old. The EEG was performed on all the patients; 20.7% had EEG abnormalities and 31.3% of these were epileptic. The non-epileptic patients presented inespecific EEG abnormalities. Therefore, our data did not allow us to propose a typical EEG pattern for DS. PMID- 10667281 TI - [Cerebrovascular disease in children: I. Epileptic manifestations]. AB - Seizures may occur as a complication of cerebrovascular disease (CVD) and its prevalence, clinical presentation, risk factors and evolution have been reported by few authors. We evaluated 39 children with CVD and analyzed the association with seizures. Seizures occurred in 24 (61.5%) patients and were classified as partial (29.2%), generalized (54.2%) and secondarily generalized (16.6%). Infants had a significantly higher prevalence of seizures (p = 0.0362) than children at other ages. Cortical localization was associated with a significantly higher prevalence of seizures (p = 0.0101). There were no differences between ischemic and hemorrhagic strokes. Fourteen patients had no seizures after the acute phase of the CVD, the 2 previously epileptic patients had their seizures controlled with antiepileptic drugs, 3 developed epilepsy, 2 died during the acute phase and in 3 patients there was not enough time yet to make a clear diagnosis of epilepsy. PMID- 10667282 TI - [Cerebrovascular disease in children: II. Clinical aspects in 42 cases]. AB - We report the findings recorded in 42 children suffering cerebrovascular disease and assisted at the Hospital das Clinicas FCM-UNICAMP, over a 8 years period (January 1990 until April 1998). The ischemic type was the most common, and involvement of the middle cerebral artery, sudden onset of clinical manifestation with seizures and motor disability were more common in early aged children. Motor sequelae predominated in the follow-up of these children. PMID- 10667283 TI - [Use of amitriptyline in attention deficit hyperactivity disorder]. AB - We studied the action of amitriptyline (AMI) in the attention deficit hyperactivity disorder (ADHD). Twenty-five children who came to consultation for ADHD were analyzed, in two groups: the group which used AMI (n = 18) at 1.6 mg/kg/day and the group which used placebo (n = 7). Both groups were submitted to two assessments in a 30 days interval, which consisted of the evolutive neurological evolution examination (ENE) and the WISC scale subtests on numbers, drawings to be completed and the code. The results showed that the AMI produced an improvement in performance in the motor persistence tests. PMID- 10667284 TI - [Intravenous lysine clonixinate for the treatment of migraine: an open pilot study]. AB - Several oral nonsteroidal anti-inflammatory drugs (NSAID) are effective to treat migraine attacks. Despite its efficacy to treat migraine and other pain, there are a few commercial NSAIDs available for intravenous (i.v.) administration. Lysine clonixinate (LC) is a NSAID derived from nicotinic acid that has been proven effective in various algic syndromes such as renal colic, nerve compression, muscular pain and odontalgias. The aim of this study was to evaluate the efficacy of the i.v. LC in the treatment of severe attacks of migraine. We studied prospectively 19 patients, 17 women and 2 men, ages from 18 to 57 years, with the diagnosis of migraine according to the International Headache Society criteria. The patients were oriented to proceed to the clinic once the headache has started, and were placed under an i.v. infusion of LC and saline in a superficial vein of the forearm, once the intensity reached severe. Evaluating the headache intensity after 30, 60 and 90 minutes, as well as the presence of side effects, we observed that all of the 19 patients were headache free after 90 minutes. Some patients presented mild adverse effects and the vital signs were not significantly affected. We then concluded that the i.v. infusion of the NSAID LC (2-3-chloro-o-toluidin)piridin-3-lysine carboxilate), a derived from the nicotinic acid with a chemical structure that resembles the flufenamic acid, was efficient abolishing a severe migraine attack after 90 minutes in 19 patients. Controlled studies with a double-blind and randomized design, and treating a greater number of patients and attacks are necessary to confirm these initial observations. PMID- 10667285 TI - [Clinical and therapeutical features in 135 patients with dystonia: experience of movement disorders unity of the Hospital de Clinicas of the Universidade Federal do Parana]. AB - This study aims to describe the clinical patterns and therapeutic responses in 135 patients with dystonia. According to the classification, 54% were focal; 17.8% were segmental; 8.1% hemidistonia; 18.6% generalized and 1.5% were multifocal. There was a positive familial history in 5.9% of the cases. The treatment of the idiopathic dystonias is divided in: specific and symptomatic, and it can be local with botulinum toxin, or systemic with oral drugs. The most common drugs used in the treatment were anticholinergics and benzodiazepines, with poor responses in the generalized forms. Botulinum toxin A was the first line treatment for focal and segmental forms of dystonia. Meanwhile, the generalized forms of dystonia show poor response to the therapies utilized. PMID- 10667286 TI - [Image guided stereotactic approach of central nervous system lesions: accuracy, morbidity, mortality]. AB - We studied seventy-five patients with brain lesions biopsied by stereotaxis from March 1993 to December 1998 at Universidade Federal de Sao Paulo-Escola Paulista de Medicina. The three most frequent lesions were: metastasis, low grade astrocytomas and glioblastoma multiforme. The morbidity rate was 2.66% due to: one case of scalp infection in a patient with thalamic cyst; and a partial seizure during surgery in a patient with lymphoma. The mortality rate was 1.33% due to increasing of cerebral edema after biopsy and the patient died after one week. The diagnostic accuracy was 89.33%. PMID- 10667287 TI - [An en bloc cranio-orbitozygomatic approach: surgical technique and results]. AB - An en bloc cranio-orbitozygomatic approach is described. This technique was applied in seven patients (five basilar artery aneurysms, a trigeminal neuroma, a meningioma of the lesser sphenoid wing). The follow-up period ranged from 3 to 50 months. The patients were retrospectively analyzed from the cosmetic point of view, and submitted to computerized tomography with three dimensional and multiplanar reconstructions. The access provided a wider operative field with a shorter distance to the pathology and possibility of increased angulation of the microscope. The aneurysms could be clipped and the tumors completely removed in all cases. A frontalis muscle paralysis occurred in two cases, as well as a temporalis muscle atrophy in another two patients. There were no enophthalmos or bone flap displacements. The surgical technique is simple and do not require drills, electric saws and mini-plates. PMID- 10667288 TI - [Stereotactic implantation of depth electrodes by magnetic resonance image in epilepsy surgery]. AB - We present the case of a 40-year-old woman with refractory epilepsy since aged 18, who was submitted to video-EEG monitoring with intracerebral depth electrodes. The clinical history and examination, magnetic resonance image (MRI), video-EEG and neuropsychological study did not allow the determination of the cerebral onset of epileptic seizures. Depth electrodes inserted by MRI-guided stereotaxis allowed the recording of the epileptic activity and thus showed quite accurately the area of the brain to be surgically resected. She underwent a right anterior temporal lobectomy with amygdalohippocampectomy. The immediate postoperative period was uneventful and she is without epileptic seizures after three months of follow-up. The average pre-operative free-seizure period was two weeks. To our knowledge, this is the first stereotactic surgery for insertion of depth intracerebral electrodes in epilepsy in Brazil. PMID- 10667289 TI - Secondary bilateral synchrony due to fronto-mesial lesions. An invasive recording study. AB - Frontal lobe epilepsies may present difficulties in focus localization in the pre operative work-up for epilepsy surgery. This is specially true in patients with normal MRIs. We report on a 16 years-old girl that started with seizures by the age of 8 years. They were brief nocturnal episodes with automatisms such as bicycling and boxing. Seizure frequency ranged from 4-10 per night. Scalp EEG showed few right frontal convexity spiking and intense secondary bilateral synchrony (SBS). High resolution MRI directed to the frontal lobes was normal. Ictal SPECT suggested a right fronto-lateral focus. Ictal video-EEG showed no focal onset. She was submitted to invasive recordings after subdural plates implantation. Electrodes covered all the frontal convexity and mesial surface bilaterally. Ictal recordings disclosed stereotyped seizures starting from the right mesial frontal. Using a high-resolution tool to measure intra and interhemispheric latencies, the timing and direction of seizure spread from the right fronto-mesial region were studied. Motor strip mapping was performed by means of electrical stimulation. She was submitted to a right frontal lobe resection, 1.5 cm ahead of the motor strip and has been seizure free since surgery (8 months). Pathological examination found a 4 mm area of cortical dysplasia. Invasive studies are needed to allow adequate localization in patients with non-localizatory non-invasive work-up and may lead to excellent results in relation to seizures after surgery. PMID- 10667290 TI - [Septic thrombosis of cavernous sinus: report of 6 cases]. AB - The cavernous sinus is most frequently involved by septic thrombosis. The common sites of primary infection are the medial face, orbits, tonsils, soft palate, sphenoid and ethmoid sinuses. The usual clinical presentation begins with fever and periorbital edema followed by headache, ptosis and ocular muscles palsy. The diagnosis is usually made on clinical grounds. Treatment consists of eradication of the primary source of infection and the administration of antibiotics and anticoagulants. We report six cases of septic thrombosis of cavernous sinus. PMID- 10667291 TI - [Prognosis factors in cryptococcal meningoencephalitis]. AB - OBJECTIVE: To identify demographic, clinical and cerebrospinal fluid (CSF) variables associated to intrahospitalar lethality of patients with cryptococcal meningoencephalitis. STUDY DESIGN: Retrospective cohort to study prognosis. SETTING: Hospital Couto Maia (HCMaia) reference for patients with infectious diseases in the State of Bahia Northeastern Brazil. POPULATION: Patients admitted at HCMaia, from 1972 to 1996, with the diagnosis of cryptococcal meningoencephalitis. RESULTS: Lethality rate was 42.7%. The most important neurological abnormalities were neck stiffness, decreased consciousness level, behavior changes, cranial nerve palsy and visual alteration. Disease time over 30 days, involvement of consciousness level and cerebrospinal fluid cells under 40/mm3 were associated to a higher lethality rate. CONCLUSION: Disease time over 30 days, involvement of consciousness level, and CSF decreased cellularity were the only predictors of lethality in the studied population. PMID- 10667292 TI - [Clinical manifestations in patients with computerized tomography diagnosis of neurocysticercosis]. AB - A survey was conducted in the urban area of Lages using patients who had been submitted to a computed tomography of the skull in the period of March-December, 1996, for different reasons. Forty-two patients with a provisional diagnosis of neurocysticercosis, and 57 negatives were personally interviewed by one of the authors (Pfuetzenreiter), using a semi-structured procedure. More individuals with a provisional diagnostic of neurocysticercosis reported clinical manifestations related to this infection than those found negative. This difference is more marked among women, except in relation to convulsions, more frequently reported by men (19.05%) than by women (7.14%). The greater percentage of inactive forms (83.33%) and a longer history of perceived symptoms among those positives suggest that the condition is not new. PMID- 10667293 TI - [True neurogenic outlet syndrome: report of 2 cases]. AB - True neurogenic thoracic outlet syndrome is caused by compression of the lower trunk of the brachial plexus usually by a cervical rib, fibrous band or an elongated transverse process of C7. We describe two cases of female patients (23 and 19 years old) with pain in the right superior limb and progressive muscular weakness and atrophy of the intrinsic muscles of hand. Electrodiagnostic studies showed reduced amplitude of compound muscle action potential of median nerve and decreased amplitude of ulnar sensory nerve action potential. Motor and sensory nerve conduction velocities were normal in both patients. Needle electromyography were findings compatible with chronic denervation in the intrinsic muscles of the right hand of both patients. Radiological investigation showed cervical ribs in one case and elongated transverse process of C7 in the other. A discussion about the clinical and electrophysiological features and the treatment of the syndrome was performed. PMID- 10667294 TI - [Type I Chiari malformation: report of 2 cases with unusual clinical presentation]. AB - We describe two patients with Chiari type I malformation with unusual clinical presentation. The first one with clinical picture of acute respiratory insufficiency and the second one with vestibular and mild cerebellar syndrome and headache. In both cases the neurological examination demonstrated the presence of "down-beating nystagmus". We emphasize the value of neurological semiology, determining a correct complementary evaluation and effective treatment. PMID- 10667295 TI - Walker-Warburg syndrome. Report of two cases. AB - The purpose of this study is to describe two infants that were diagnosed with Walker-Warburg syndrome (WWS), a rare form of congenital muscular dystrophy (CMD). They were studied in their clinical, laboratory, and neuroradiologic features. The index case had a brain magnetic resonance imaging (MRI) and the second patient had a head computerized tomography (CT). In addition, a literature review was performed to describe the main forms of CMD. The index case fulfilled all criteria for WWS. A brain MRI performed at age 4 months served to corroborate the clinical diagnosis, showing severe hydrocephalus, type II lissencephaly, cerebellar vermian aplasia, and a hypoplastic brain stem. The authors were able to establish a retrospective diagnosis of WWS in the index case's older sister, based upon her clinical picture and head CT report. PMID- 10667296 TI - Pseudocystic form of neurocryptococcosis in pregnancy. Case report. AB - We report a case of neurocryptococcosis which is unique in the literature because the patient had a pseudocystic form of the disease during pregnancy and without any evidence of AIDS. The clinical picture was that of intracranial hypertension and the epidemiological background was highly suggestive of cysticercosis. CT showed multiple round hypodense lesions in the basal ganglia and cerebellum, without contrast enhancement. Since a scolex was not visible, the diagnosis of neurocysticercosis was considered probable. CSF examination was not performed in view of its high risk. The patient had progressive downhill course. Autopsy disclosed multiple gelatinous pseudocysts in the cerebral and cerebellar gray matter, containing abundant Cryptococcus neoformans. Meningeal involvement was minimal. The child was delivered by caesarean section and was free of infection, but died later of hyaline membrane disease. The neuroimaging appearances of this rare instance of the pseudocystic form of neurocryptococcosis mimicked closely neurocysticercosis and only postmortem examination allowed correct diagnosis. The pseudocystic form has so far only been reported in AIDS. PMID- 10667297 TI - Treatment of cervical dystonia with botulinum toxin in a patient with myasthenia gravis. AB - We report the case of a 49-year-old woman who has the rare combination of myasthenia gravis and cervical dystonia. She was treated with botulinum toxin type A with good response and no evidence of deterioration of the myasthenic symptoms. We therefore conclude that it is possible to use botulinum toxin in the presence of defective neuromuscular transmission. PMID- 10667298 TI - [Identical twins discordant for Cushing's disease: case report[]. AB - Cushing's disease is rare in children and its occurrence in identical twins is extremely rare. This paper reports on identical twins discordant for Cushing's disease. One of them first presented with a cushingoid phenotype by the age of 10. Her evaluation showed an increased urinary free-cortisol and serum ACTH. Her pattern in the dexametazone suppression tests was compatible with Cushing's disease. MRI disclosed a pituitary macroadenoma which was removed by the transesphenoidal approach. Immunohistochemical studies of the tumor showed the presence of ACTH-producing cells. The patient went into clinical and laboratorial remission after surgery. She re-started to grow after the disappearance of the Cushing's phenotype but she is still shorter than her healthy sister. The latter remains disease-free 4 years after her sister's diagnosis. This represents the third such case reported in the literature. Our findings suggest that acquired factors may be responsible for the genesis of Cushing's disease. PMID- 10667299 TI - [Actinomycosis of the brain: case report]. AB - Actinomycosis located in the central nervous system is an extremely uncommon event, but if correctly diagnosed and properly treated may have a good prognosis. This case report of a cerebral abscess caused by actinomyces suggests that such a rare event should be included in the differential diagnosis of infectious diseases that affect the central nervous system. PMID- 10667300 TI - [Emotionally-loaded narrative increases dyslogia in schizophrenics]. AB - We tested the degree of dyslogia in the narrative of ten schizophrenic non medicated outpatients while they narrated emotionally-loaded or neutral facts. The "emotional narratives" were much more dyslogic than the "neutral narratives". In order to explain these facts we evoke: 1. A fronto-temporo-limbic connection dysfunction would disturb an adequate cognitive treatment of emotions that would be, in this way, highly disruptive over logical processes. 2. A working memory/supervisory attentional system dysfunction that would produce both a loss of the normal connections among the fragments of speech and a lack of global strategical planning of the thought. PMID- 10667301 TI - Sex playing with the mind. Effects of oestrogen and testosterone on mood and cognition. AB - Women now spend more than 1/3 of their lives in a state of oestrogen deprivation as a result of increased life expectancy. A similar, but milder, hypogonadal state has been described for elderly men. This paper aims to review the available literature on the effects of both oestrogen and testosterone on mood and cognition. Oestrogen replacement therapy of postmenopausal women is associated with improvements in measures of well being and decline in depression scores. In addition, oestrogen seems to augment the response of postmenopausal women with major depression to antidepressant treatment. Most studies designed to investigate the impact of oestrogen on cognition indicate that replacement therapy is associated with better performance on neuropsychological tests, particularly in measures of verbal memory and fluency. The data also supports claims that oestrogen replacement therapy reduces the risk of Alzheimer's disease in later life and improves response of patients to anticholinesterase treatment. Data on the effects of testosterone is sparser. Preliminary findings suggest that testosterone therapy may improve mood when used in isolation or in association with oestrogen. The effects of testosterone on cognitive functioning are less clear--some studies indicate that the administration of testosterone to non demented subjects is associated with better visuospatial functioning and deterioration of verbal skills. In summary, gonadal hormones seem to modulate various aspects of mental functioning. If future studies prove this to be true, hormone replacement therapy should have a major impact on the physical and mental health of older people in the years to come. PMID- 10667302 TI - Function and regulation of prostaglandin synthase 2. PMID- 10667303 TI - Regulation of cyclooxygenase-2 by the activated p38 MAPK signaling pathway. PMID- 10667304 TI - Fatty acid cyclooxygenase induction and prostaglandin D synthesis in a human megakaryoblastic cell line CMK differentiated by phorbol ester. PMID- 10667305 TI - Eicosanoid metabolism in human platelets is modified by albumin. PMID- 10667306 TI - Diverse functional coupling of prostanoid biosynthetic enzymes in various cell types. AB - As also detailed in our accompanying papers in this issue, recent studies have revealed functional crosstalk and segregation between PLA2s, COXs, and terminal PG synthases in various cells (Table I). Among the PLA2s, cPLA2 is required for all three responses, and sPLA2-IIA augments the delayed response in preference to the immediate response. sPLA2-IIA associates with proteoglycans on the surface of stimulus-primed cells to exert its functions. COX-1 is utilized only in the immediate response and COX-2 is a prerequisite for the delayed response. The induced immediate response is often mediated by COX-2 rather than by COX-1, especially when the end product is PGE2. In addition to segregated utilization of these enzymes, significant crosstalk and/or synergism between them, which is often cell type specific, is also obvious. For instance, sPLA2 acts as an enhancer of COX-2 expression in rat mast cells, functional cPLA2 is required for sPLA2 induction in rat fibroblasts, and sPLA2 augments cPLA2 and COX-2 expression in mouse osteoblasts via endogenous PGE1. Moreover, differential coupling between COXs and downstream terminal PG synthases is also evident in macrophages, in which COX-1 and COX-2 are preferentially coupled with TXS and PGES, respectively. Thus, different PG-biosynthetic enzymes, acting on different cellular AA pools at different locations and being regulated by separate but interacting mechanisms, confer on the system great versatility in ensuring that both immediate and delayed AA-derived mediators are efficiently generated during cellular responses. PMID- 10667307 TI - Regulation of prostaglandin, leukotriene, and platelet-activating factor metabolism in mast cells. PMID- 10667308 TI - Lipid peroxides and neuronal plasticity. PMID- 10667309 TI - Secretion of lipocalin-type prostaglandin D synthase (beta-trace) from human heart to plasma during coronary circulation. PMID- 10667310 TI - Lipocalin-type prostaglandin D synthase (beta-trace) binds non-substrate lipophilic ligands. PMID- 10667311 TI - Features of mammalian lipoxygenases. PMID- 10667312 TI - Identification and characterization of an enhancer sequence in the promoter region of human 15-lipoxygenase (15-LO) gene. PMID- 10667313 TI - Cytokine induced regulation of 15-lipoxygenase and phospholipid hydroperoxide glutathione peroxidase in mammalian cells. PMID- 10667314 TI - Investigation of a second 15S-lipoxygenase in humans and its expression in epithelial tissues. PMID- 10667315 TI - Sequence determinants for the positional specificity of mammalian and plant lipoxygenases. PMID- 10667316 TI - X-ray absorption studies into the iron ligand sphere of plant and animal lipoxygenases. PMID- 10667317 TI - Transcriptional and posttranscriptional regulation of 5-lipoxygenase mRNA expression in the human monocytic cell line Mono Mac 6 by transforming growth factor-beta and 1,25-dihydroxyvitamin D3. PMID- 10667318 TI - Comparison of prostaglandin H synthase-1 and -2 structural stabilities. PMID- 10667319 TI - Modulation of the expression of the cyclooxygenase 1 and 2 genes in rat mammary glands: role of hormonal status and dietary fat. PMID- 10667320 TI - Regulation of cerebrovascular cyclooxygenase-2 by pro- and anti-inflammatory cytokines. PMID- 10667321 TI - Visualization and quantitation of cyclooxygenase-1 and -2 activity by digital fluorescence microscopy. PMID- 10667322 TI - Discovery of a new class of selective cyclooxygenase-2 (COX-2) inhibitor that covalently modifies the isozyme. PMID- 10667323 TI - Mitigation of arthritis by high-dose administration of a COX-2 inhibitor in the collagen-induced arthritis model in the mouse. PMID- 10667324 TI - Indomethacin inhibition of pristane plasmacytomagenesis in genetically susceptible inbred mice. PMID- 10667325 TI - Role of COX-2 inhibition on the formation and healing of gastric ulcers induced by indomethacin in the rat. PMID- 10667326 TI - Regulation of in vivo prostaglandin biosynthesis by glutathione. PMID- 10667327 TI - Different effects of reactive nitrogen intermediates on prostaglandin E2 synthesis in cultured rat microglia and RAW 264.7 cells. PMID- 10667328 TI - Activation of cPLA2 in vascular smooth muscle. PMID- 10667329 TI - Role of type IIA secretory phospholipase A2 in arachidonic acid metabolism. AB - Recent recognition of the rapidly growing sPLA2 family has led to a suggestion that some of the previously described functions of sPLA2-IIA need to be reevaluated, since studies based upon enzyme activities and using inhibitors or antibodies against sPLA2-IIA may not discriminate these sPLA2s. Our present studies reconfirm the involvement of sPLA2-IIA in biological responses, demonstrated significant crosstalk between the two Ca(2+)-dependent PLA2s (cPLA2 and sPLA2) where one enzyme is required for the induction of the other, and revealed segregated coupling of discrete PLA2 and COX enzymes in the different phases of PG biosynthesis. Based upon the analysis of cells derived from sPLA2 IIA "natural knock-out" mice, it is apparent that sPLA2-IIA is not essential for the initiation of delayed PGE2 biosynthesis. However, it is capable of contributing to the delayed response as an enhancer when appropriately induced by proinflammatory stimuli, leading to optimal COX-2-dependent PGE2 generation. Importantly, in order for sPLA2-IIA (or related sPLA2 isozymes) to attack the biological membranes, so-called "membrane rearrangement" should take place in activated, but not resting, cells. Membrane rearrangement also occurs when cells are undergoing apoptosis, during which acidic phospholipids, the preferred substrates for sPLA2-IIA, are exposed on the outer leaflet of the plasma membranes. Nonetheless, in view of the dramatically elevated levels of sPLA2-IIA in inflamed or ischemic sites, it is likely that this extracellular isozyme participates in the expansion of chronic tissue disorders by augmenting generation of proinflammatory eicosanoids or lysophospholipids, depending upon the states of the inflammatory response. PMID- 10667330 TI - Phospholipase A2 is involved in chemotaxis of human leukocytes. PMID- 10667331 TI - Suppression of acute experimental inflammation by antisense oligonucleotides targeting secretory phospholipase A2 (sPLA2) in vitro and in vivo experiments. AB - In HepG2 cells phosphorothioate modified antisense oligonucleotides against a sequence in the Ca2+ binding domain (AS-Ca2+) of type II sPLA2 mRNA restrained IL 6-induced synthesis of sPLA2 protein, sPLA2 mRNA (northern blot), and abolished IL-6 stimulated PGE2 release. An antisense oligonucleotide corresponding to a sequence in the catalytic domain (AS-Cat) of sPLA2 was less effective. The antisense oligonucleotides did not affect albumin synthesis in HepG2 cells, additionally demonstrating their specificity. The corresponding AS-Ca2+ against a homologous part of the rat sPLA2 mRNA depressed rat carrageenin oedema for 60 70%. Identical suppression was achieved by specific low molecular weight inhibitors of sPLA2. Since cyclo- and 5-lipoxygenase inhibitors exerted similar reductions of carrageenin oedema type II sPLA2 dependent eicosanoid formation seems to be a key cascade in this type of inflammation. PMID- 10667332 TI - Comparison of recombinant types IIA, V and IIC phospholipase A2S, the three related mammalian secretory phospholipase A2 isozymes. PMID- 10667333 TI - Respective roles of the 14 kDa and 85 kDa phospholipase A2 enzymes in human monocyte eicosanoid formation. AB - Human monocytes possess both the cytosolic 85 kDa phospholipase (PLA) A2 and a 14 kDa PLA2 and are capable of simultaneously producing prostanoids (PG), leukotrienes (LT) and platelet activating factor (PAF). As the exact roles of the two enzymes in monocyte lipid mediator formation was unclear, both selective PLA2 inhibitors and antisense were used to elucidate their respective roles. Reduction in 85 kDa PLA2 cellular protein levels by initiation site-directed antisense (SK 7111) or exposure to the 85 kDa PLA2 inhibitor, arachidonyl trifluormethyl ketone (AACOCF3), prevented A23187 or zymosan-stimulated monocytes prostanoid formation but not LTC4 or PAF production. This confirmed the important role of the 85 kDa PLA2 in prostanoid formation but indicated a less significant role in LT or PAF biosynthesis. Alternatively, treatment of monocytes with the selective, active site-directed 14 kDa PLA2 inhibitor, SB 203347, totally inhibited LT and PAF formation, while prostanoid formation was not altered. Addition of 20 uM exogenous arachidonic acid (AA) to monocytes exposed to SB 203347 did not alter A23187-induced LTC4 generation, indicating that SB 203347 had no effect on downstream AA metabolizing enzymes in this setting. Taken together, these results provide evidence that the 14 kDa PLA2 provides substrate for monocyte LT and PAF formation, while the 85 kDa PLA2 plays a more significant role in the formation of PG. PMID- 10667334 TI - Modulation of long-term potentiation in the CA1 area of rat hippocampus by platelet-activating factor. PMID- 10667335 TI - PPARs: nuclear receptors for fatty acids, eicosanoids, and xenobiotics. AB - The peroxisome proliferator-activated receptors have enjoyed the spotlight for many reasons. These transcription factors are ligand-inducible nuclear receptors that modulate gene expression in response to a broad spectrum of compounds. The recognition that PPARs are indeed nuclear receptors for polyunsaturated fatty acids, some eicosanoids and also lipid-lowering and antidiabetic drugs, has opened many exciting avenues of research and drug discovery. Recent studies on the PPAR function have extended the role of these transcription factors beyond energy homeostasis to master gene in adipogenesis and also determinants in inflammation control. While rapid advances have been made, it is clear that we are far from a global understanding of the mechanisms and functions of PPARs. PMID- 10667336 TI - Molecular cloning and characterization of leukotriene B4 receptor. PMID- 10667337 TI - Determinants of receptor subtype specificity in the LPA-like lipid mediator family. PMID- 10667338 TI - Subunits and cellular occurrence of the 12(S)-HETE binding complex. PMID- 10667339 TI - A subfamily of G protein-coupled cellular receptors for lysophospholipids and lysosphingolipids. AB - The results of molecular cloning and homology searches have identified a minimum of five different proteins of the endothelial differentiation gene (edg) encoded subfamily of GPCRs. Edg protein GPCRs show amino acid sequence identity of 31% to 34% as a subfamily, but contain two homology clusters with greater similarity of structures and functions. One cluster of high amino acid sequence homology includes Edg-2 and Edg-4 proteins, that encode GPCRs for LPA, but not lysosphingolipids. A second homology cluster encompasses Edg-1, Edg-3 and Edg-5. Edg-3 and Edg-5 encode GPCRs specific for S1P, but not LPA. Preliminary data suggest that Edg-1 encodes a GPCR for S1P and one or more other lysosphingolipids, but the signals evoked by S1P alone are far weaker than those transduced by Edg-3 and Edg-5. Similarities of the structures of genes for the respective homology clusters supports this tentative classification of the Edg protein GPCRs. Future research will be directed to completion of the elucidation of genomic organization and signaling pathways, and a greater understanding of the breadth of functional roles of Edg proteins in development and activities of the nervous, cardiovascular, endocrine and immune systems. PMID- 10667340 TI - Evidence for involvement of prostaglandin I2 as a major nociceptive mediator in acetic acid-induced writhing reaction: a study using IP-receptor disrupted mice. PMID- 10667341 TI - Local anesthetic effects on TXA2 receptor mediated platelet aggregation using quenched flow aggregometry. PMID- 10667342 TI - Volatile and local anesthetics interfere with thromboxane A2 receptors recombinantly expressed in Xenopus oocytes. PMID- 10667343 TI - Aspirin-triggered 15-epi-lipoxin A4 and novel lipoxin B4 stable analogs inhibit neutrophil-mediated changes in vascular permeability. PMID- 10667344 TI - Cleavage of leukotriene D4 in mice with targeted disruption of a membrane-bound dipeptidase gene. PMID- 10667345 TI - Gamma-glutamyl leukotrienase cleavage of leukotriene C4. PMID- 10667346 TI - LTE4 blood levels in infants with congenital heart lesions. PMID- 10667347 TI - Evaluation of the pharmacological activity of the pure cysteinyl-leukotriene receptor antagonists CGP 45715A (iralukast) and CGP 57698 in human airways. PMID- 10667348 TI - Evidence for a carbocation intermediate in the enzymatic transformation of leukotriene A4 into leukotriene B4. PMID- 10667349 TI - A random rapid equilibrium mechanism for leukotriene C4 synthase. AB - Kinetic studies performed on the conjugation reaction catalyzed by LTC4 synthase proved to conform to a random rapid equilibrium mechanism which was further substantiated by competition patterns ruling out other possible mechanisms. Most cytosolic Gst's investigated to date appear to follow a random kinetic mechanism although are mainly responsible for detoxification purposes. Conversely, LTC4 synthase possesses a very different biological role yet still follows a similar mechanism. Therefore, it can be concluded that most GSTs function in a consistent manner regardless of their biological function. Of interest are the mechanisms of the other members of the MAPEG family which in some respects are closer to conventional GSTs than to LTC4 synthase, yet they remain to be deciphered. PMID- 10667350 TI - Formation of reactive products of the isoprostane pathway: isolevuglandins and cyclopentenone isoprostanes. PMID- 10667351 TI - Formation of novel isoprostane-like compounds from docosahexaenoic acid. PMID- 10667352 TI - cDNA cloning, expression and chromosomal localization of two human lysophosphatidic acid acyltransferases. AB - In this report we describe a pair of human LPAAT isozymes. These isozymes are encoded by distinct genes located on different chromosomes, but share sequence homology, substrate specificity, and intracellular location. The biological value of maintaining the two closely related LPAAT genes in the human genome is not clear. We find that both isozymes are widely expressed, although expression levels do diverge significantly in tissues such as the liver, placenta, testes, and pancreas. We also find that, at least in the artificial system of over expression in COS7 cells, both isozymes localize to the ER membrane. Thus, distinct tissue-specific or subcellular compartment-specific roles for the two isozymes are not supported by the current experimental evidence. It does remain possible that induction of expression or subcellular translocation of one or the other isozyme may distinguish their functions. A survey of a limited number of acyl CoA substrates indicates that the two isozymes display similar substrate specificities, although slight differences are suggested by the data. However, extensive analysis of both isozymes with multiple substrates in the same assay system will be required to detect physiologically relevant differences in substrate specificity. LPA and PA are central intermediates in phospholipid biogenesis. Furthermore, they have the capacity to mediate signaling both between and within cells. The importance of these mediators is reflected in the growing body of literature dedicated to unraveling the mechanistic basis for their actions. Until recently, the field has been hampered by a dearth of reagents appropriate for the molecular dissection of the LPA and PA metabolic and signaling pathways in eukaryotes. However, the recent cloning of possible LPA receptors will promote further understanding of LPA signaling. Similarly, the recent appearance of LPAAT homologs in the EST database has prompted a flurry of reports describing their characterization. These clones will afford opportunity for defining the function of LPAAT in eukaryotic phospholipid metabolism. PMID- 10667353 TI - The first cloned and identified lysophospholipid (LP) receptor gene, vzg-1: implications for related receptors and the nervous system. PMID- 10667354 TI - Involvement of protein kinase C, p38 MAP kinase and ERK in arachidonic acid stimulated superoxide production in human neutrophils. PMID- 10667355 TI - 13(S)-HpODE augments epidermal growth factor signal transduction by attenuating EGF receptor dephosphorylation. PMID- 10667356 TI - Role of lipoxygenase in the regulation of glucose transport in aortic vascular cells. PMID- 10667357 TI - Functions of the p21-activated protein kinases (Paks) in neutrophils and their regulation by complex lipids. PMID- 10667358 TI - Inhibition of neurofibromin and p120 GTPase activating protein (GAP) by dietary fatty acids. AB - Neurofibromin and p120 GTPase activating protein (p120 GAP) down-regulate the activity of cellular Ras proteins. How the activity of these two proteins is controlled is not yet clear. In this study, we analyzed the effects of eight nutritionally relevant fatty acids on GTPase stimulatory activity of full-length neurofibromin and p120 GAP. The fatty acids tested were: saturated stearic acid, monounsaturated oleic acid, three omega-6 and three omega-3 polyunsaturated fatty acids. The analysis was performed by Ras immunoprecipitation GTPase assay. The full-length p120 GAP expressed in insect Sf9 cells and the immunoaffinity purified full-length neurofibromin were used. Neurofibromin was readily inhibited by stearic and oleic acid, but p120 GAP was not inhibited even at high concentrations (> 80 microM). Neurofibromin was also inhibited by low concentrations of all the polyunsaturated fatty acids tested (IC50 of 6 to 16 microM). p120 GAP was 2-3 fold less sensitive to inhibition by these fatty acids. The GTPase stimulatory activity of neurofibromin was also inhibited by arachidonic and oleic acid in the presence of a lipid mixture representing the major lipid components of the cell membrane. Chimeric proteins of neurofibromin and p120 GAP were used to determine that differential sensitivity to fatty acid inhibition maps to the catalytic domain of the proteins. These results indicated that fatty acids can modulate the GTPase function of the c-Ha-Ras protein by inhibiting the GTPase stimulatory activity of two Ras regulators, full-length neurofibromin and p120 GAP, at physiologically relevant concentrations in vitro. PMID- 10667359 TI - Spinal synthesis and release of prostanoids after peripheral injury and inflammation. PMID- 10667360 TI - Prostaglandin E2 as a modulator of lymphocyte mediated inflammatory and humoral responses. PMID- 10667361 TI - Lipid mediators stimulate reactive oxygen species formation in immortalized human keratinocytes. PMID- 10667362 TI - Nitric oxide synthesis is increased in periodontitis. PMID- 10667363 TI - Augmentation effects of high glucose on endotoxin-induced nitric oxide production in murine macrophages. PMID- 10667364 TI - 1,2-Diacylglycerol hydroperoxide induces the generation and release of superoxide anion from human polymorphonuclear leukocytes. PMID- 10667365 TI - Kinetic evaluation of endogenous leukotriene B4 and E4 acute activation of inflammatory cells in the rabbit. PMID- 10667366 TI - Lipopolysaccharide- and liposome-encapsulated MTP-PE-induced formation of eicosanoids, nitric oxide and tumor necrosis factor-alpha in macrophages. PMID- 10667367 TI - LTA4 hydrolase expression during glomerular inflammation: correlation of immunohistochemical localization with cytokine regulation. PMID- 10667368 TI - 12-Lipoxygenase products increase monocyte:endothelial interactions. AB - In summary, we suggest that hyperglycemia causes upregulation of 12-lipoxygenase activity. The increased production of 12-LO products, 12(S) and 15(S)-HETE, activates monocyte integrins which result in enhanced adhesion of monocytes to endothelium. The binding of monocytes to endothelium is a key early event in development of atherosclerosis. Upregulation of this process by vascular cells exposed to chronic elevations in glucose may be one explanation for the accelerated atherosclerosis observed in patients with Type 2 diabetes. PMID- 10667369 TI - Linoleic acid metabolites in health and disease. PMID- 10667370 TI - Toxicity of linoleic acid metabolites. PMID- 10667371 TI - Dramatic increase of linoleic acid peroxidation products by aging, atherosclerosis, and rheumatoid arthritis. PMID- 10667372 TI - Modulation of atherogenesis by dietary gamma-linolenic acid. AB - Data from our in vitro studies indicate that macrophages isolated from mice fed GLA-enriched diets inhibit vascular SMC proliferation via a PGE1-cAMP dependent mechanism. Since SMC proliferation is one of the main events implicated in the pathogenesis of atherosclerosis (Ross, 1993), this anti-proliferative effect observed by dietary GLA is noteworthy. In vivo studies have established that dietary GLA is capable of retarding the atherosclerotic lesion formation in ApoE knock out mice, an animal model that develops atherosclerosis similar to humans (Reddick, 1994). We propose that dietary GLA has the potential to inhibit SMC proliferation leading to retardation of atherosclerotic lesion formation, and therefore favorable modulation of the atherogenic process. PMID- 10667373 TI - The selective cytotoxicity of gamma-linolenic acid (GLA) is associated with increased oxidative stress. PMID- 10667374 TI - gamma-linolenic acid (GLA)-mediated cytotoxicity in human prostate cancer cells. PMID- 10667375 TI - Five-lipoxygenase inhibitors reduce Panc-1 survival: synergism of MK886 with gamma linolenic acid. PMID- 10667376 TI - Enzymological and molecular biological studies on anandamide amidohydrolase. AB - Previously we suggested that anandamide amidohydrolase partially purified from porcine brain catalyzed the anandamide synthesis. The reversibility of the anandamide hydrolytic reaction was confirmed with a recombinant enzyme of rat liver. We also showed that the recombinant enzyme had a wide substrate specificity hydrolyzing primary amides and esters of fatty acids in addition to anandamide. When the organ distribution of anandamide amidohydrolase was examined with rats, a large amount of the enzyme was contained in small intestine as well as liver and brain. The intestinal hydrolase was masked by endogenous lipid inhibitors. The enzyme was also found in various eye tissues. PMID- 10667377 TI - A pathway for the biosynthesis of fatty acid amides. PMID- 10667378 TI - Investigation of structural analogs of prostaglandin amides for binding to and activation of CB1 and CB2 cannabinoid receptors in rat brain and human tonsils. PMID- 10667379 TI - Hepoxilin A3 is metabolized into its omega-hydroxy metabolite by human neutrophils. PMID- 10667380 TI - Oxidation of arachidonate containing glycerophospholipids in intact red blood cells and red blood cell membranes with tert-butylhydroperoxide. PMID- 10667381 TI - Cannabinoid modulation of neuronal activity in adult rat hippocampus. PMID- 10667382 TI - Cyclooxygenase-independent induction of p21WAF-1/CIP1, apoptosis and differentiation by L-745,337 and salicylate in HT-29 colon cancer cells. PMID- 10667383 TI - Modulation of cellular proliferation and induction of apoptosis in a human lymphoma cell line after treatment with selective lipoxygenase inhibitors. PMID- 10667384 TI - Mechanisms of peroxynitrite-induced apoptosis in HL-60 cells. PMID- 10667385 TI - Central role of arachidonate 5-lipoxygenase in the regulation of cell growth and apoptosis in human prostate cancer cells. PMID- 10667387 TI - The possible involvement of 15-lipoxygenase/leukocyte type 12-lipoxygenase in colorectal carcinogenesis. PMID- 10667386 TI - Role of autocrine motility factor in a 12-lipoxygenase dependent anti-apoptotic pathway. PMID- 10667388 TI - Induction of leukotriene B4 metabolism by cancer chemopreventive agents. PMID- 10667389 TI - Intestinal tumor load in the Min/+ mouse model is not correlated with eicosanoid biosynthesis. PMID- 10667390 TI - 12-lipoxygenase expression in human melanoma cell lines. AB - 12-lipoxygenase (12-LOX) expression and function in the regulation of the metastatic phenotype was demonstrated in several murine melanoma lines before. Here we have provided novel evidences that, though at a low level (in max. 15% of the cell population), human melanoma lines (HT168, M1, HT199, HT18 and WM35) express the platelet-type isoform of 12-LOX both at mRNA and protein levels. 12 LOX expression was demonstrated in cultured tumor cells and in skin tumor xenografts. Comparison of the expression of 12-LOX in skin primary tumors and its lung metastases indicated a stable expression. The low level of 12-LOX expression in human melanoma cell lines suggests that other lipoxygenase(s) could also be responsible for the metabolism of arachidonic acid to 12-HETE breakdown products. PMID- 10667391 TI - Platelet-type 12-lipoxygenase regulates angiogenesis in human prostate carcinoma. PMID- 10667392 TI - Prostaglandin H synthase and lipoxygenase mediated activation of xenobiotics in platelets. AB - To investigate the involvement of prostaglandin H synthase (PHS) and lipoxygenase (LPO) in the activation of xenobiotics in platelets, platelet sonicates were preincubated with alpha-naphthol. Protein covalent binding of alpha-naphthol was measured following addition of arachidonic acid. Protein covalent binding was increased in a dose-dependent manner until it plateaued at 500 microM arachidonic acid. Pretreatment by two inhibitors of PHS, aspirin and indomethacin, resulted in a dose-dependent inhibition of alpha-naphthol-induced covalent binding, confirming PHS involvement. In addition, pretreatment by a LPO inhibitor, nordihydroguaiaretic acid (NDGA), also prevented covalent binding substantially, showing that LPO may be an alternative pathway for xenobiotic activation in platelets. Furthermore, combined treatment of aspirin and NDGA almost abolished the increase of alpha-naphthol-induced covalent binding, suggesting that PHS and LPO are both major pathways for xenobiotic activation in platelets. PMID- 10667393 TI - Characterization of two spliced variants of human phosphatidic acid phosphatase cDNAs that are differentially expressed in normal and tumor cells. PMID- 10667394 TI - Eicosapentaenoic acid alters manganese superoxide dismutase immunoreactive protein levels in normal but not malignant central nervous system derived cells. PMID- 10667395 TI - Gamma radiation and release of norepinephrine in the hippocampus. PMID- 10667396 TI - (3R)-hydroxy-oxylipins--a novel family of oxygenated polyenoic fatty acids of fungal origin. PMID- 10667397 TI - Eicosanoids in the brain of warm- and cold-acclimated bullfrogs. PMID- 10667398 TI - Production of 3-hydroxy fatty acids by the yeast Dipodascopsis uninucleata. Biological implications. PMID- 10667399 TI - Catalytic properties of linoleate diol synthase of the fungus Gaeumannomyces graminis: a comparison with PGH synthases. PMID- 10667400 TI - Carboplatin. PMID- 10667401 TI - The accuracy of intraoperative cytopathological diagnosis compared with conventional histopathological diagnosis. AB - To determine the accuracy of intraoperative cytopathological diagnosis compared with conventional histopathological diagnosis, the authors obtained 100 specimens from masses of various organ systems chosen randomly from 65 dogs, 30 cats, and five exotic animals. Of the 100 specimens, a specific diagnosis was obtained in 42%, the correct pathological process (i.e., mesenchymal neoplasia, epithelial neoplasia, round cell neoplasia, or inflammation) was identified in 41%, in 1% the diagnosis was deferred, and in 16% an incorrect diagnosis was obtained. The overall accuracy rate of intraoperative cytopathological examination was 83%, which increased to 90% by the exclusion of splenic masses. The accuracy rate of diagnosing neoplasia was 87%, with a sensitivity of 89% and a specificity of 100%. Intraoperative cytopathological examination is an accurate diagnostic method with good sensitivity and specificity for the identification of neoplasia. PMID- 10667402 TI - Hypercalcemia in a dog: a challenging case. AB - An 18-month-old, spayed female, mixed-breed dog was referred for investigation of persistent hypercalcemia. After extensive diagnostic evaluation, a tentative diagnosis of occult lymphosarcoma (LSA) was made and the dog was euthanized. At necropsy, infection with Heterobilharzia americana was diagnosed. In endemic areas, schistosomiasis should be included in the differential diagnosis of hypercalcemia, and a fecal examination should be performed in every dog with a hypercalcemia of unknown origin. PMID- 10667403 TI - Glucagon constant-rate infusion: a novel strategy for the management of hyperinsulinemic-hypoglycemic crisis in the dog. AB - A six-year-old, spayed female, cocker spaniel was presented for hypoglycemic seizures. Hypoglycemia with concomitant hyperinsulinemia suggested an insulin secreting tumor. Pancreatic masses were resected, and insulinoma was diagnosed. Six weeks later, the dog presented in hyperinsulinemic-hypoglycemic crisis (HHC). The dog was initially stabilized with intravenous dextrose boluses and infusions, but seizure activity recurred and persisted. A glucagon constant-rate infusion (GCRI) was initiated, and neurological signs quickly resolved. Dextrose was withdrawn over 24 hours, and euglycemia was maintained by GCRI alone. Despite aggressive medical management including the use of prednisone, diazoxide, bovine somatotropin, and streptozocin, the dog presented on two subsequent occasions in HHC and both times was rapidly stabilized with GCRI alone. In this dog, GCRI was a fast, safe, and effective method of achieving and maintaining euglycemia despite intractable hyperinsulinism. The clinical use of GCRI merits further investigation for management of HHC in veterinary species. PMID- 10667404 TI - Syringohydromyelia in Cavalier King Charles spaniels. AB - Syringohydromyelia secondary to foramen magnum overcrowding is described in seven Cavalier King Charles spaniels. Clinical signs were consistent with a central spinal cord lesion. The most common signs were persistent scratching at the shoulder region with apparent neck, thoracic limb, or ear pain and thoracic limb lower motor neuron deficits. The diagnosis was made by magnetic resonance imaging. The syringohydromyelia is postulated to be a consequence of an occipital bone malformation resulting in a small caudal fossa and cerebellar herniation. Clinical signs improved but did not completely resolve when the dogs received treatment with corticosteroids or nonsteroidal anti-inflammatory drugs. PMID- 10667405 TI - A modified bilateral transfrontal sinus approach to the canine frontal lobe and olfactory bulb: surgical technique and five cases. AB - Five adult dogs presented for an acute onset of seizure activity. Magnetic resonance imaging revealed lesions in the olfactory bulbs, frontal lobes of the cerebrum, or both. A modified bilateral transfrontal sinus craniotomy was performed on each patient. The goal of removing the lesion was to relieve clinical signs and to provide tissue for histopathological diagnosis. In each instance, excision of the lesion was possible using this approach. No postoperative complications were observed. The modified bilateral transfrontal sinus craniotomy provides excellent access to the canine olfactory bulbs and frontal lobes. PMID- 10667406 TI - Probable hypercalcemia of malignancy in a cat with bronchogenic adenocarcinoma. AB - An eight-year-old, neutered male, domestic shorthair cat was referred with a four day history of acute vomiting. Hypercalcemia was identified on serum biochemical testing. Thoracic radiographs showed multiple pulmonary nodular densities. Postmortem and histopathological examination identified the nodules as bronchogenic adenocarcinoma with metastases to the tracheobronchial lymph nodes, diaphragm, and parietal pleura. To the authors' knowledge, this is the first reported case of hypercalcemia of malignancy associated with bronchogenic adenocarcinoma in a cat. PMID- 10667407 TI - Acute bilateral trigeminal neuropathy associated with nervous system lymphosarcoma in a dog. AB - A nine-year-old dog presented with clinical signs consistent with bilateral trigeminal neuropathy. Multicentric lymphoma was diagnosed, and neoplastic lymphocytes were identified in the cerebrospinal fluid. Electromyography revealed spontaneous activity in temporal and masseter muscles. Histopathological examination demonstrated neoplastic cell invasion of temporal and masseter myofibers and of multiple peripheral nerves, including the trigeminal nerve. Central nervous system pathology consisted primarily of spinal root and leptomeningeal lymphoid cell infiltration with relative sparing of spinal cord and brain parenchyma. PMID- 10667408 TI - Skeletal muscle lymphoma in a bullmastiff. AB - A 16-month-old, neutered male bullmastiff was presented for acute onset of massive swelling of the right hind limb. Primary skeletal muscle lymphoma was diagnosed based on cytopathology, surgical biopsy, and necropsy findings. Cutaneous metastases developed during the hospitalization, and additional metastases were found in the heart and thoracic wall. Primary skeletal muscle lymphoma is a rare form of lymphoma in dogs and should be considered as a differential diagnosis for acute, soft-tissue swelling of the limb. PMID- 10667409 TI - Caudal cervical intervertebral disk disease in the small dog: role of distraction and stabilization in ventral slot decompression. AB - The clinical outcomes in 112 dogs weighing less than 35 pounds that were presented with cervical intervertebral disk protrusions were retrospectively evaluated. Although the second to third cervical (C2 to C3) intervertebral space was the most common site (27%) of disk protrusion, 57% of disk protrusions presented were caudal to the fourth cervical (C4) vertebra. Dogs with cranial intervertebral disk protrusions, including the C2 to C3 and C3 to C4 intervertebral disk spaces, responded favorably to ventral slot decompression. By comparison, caudal intervertebral disk protrusions (within the C4 to the seventh cervical [C7] intervertebral disk spaces) responded less favorably to ventral slot decompression, demonstrating significantly more severe clinical effects in motor function, comfort, recovery, and long-term outcome following surgery. Significant improvement in clinical results was seen in caudal disk protrusions when additional surgical distraction and stabilization were provided following ventral slot decompression. PMID- 10667410 TI - Use of a free cortical ulnar autograft following en bloc resection of a mandibular tumor. AB - A dog was presented for the en bloc resection of a previously irradiated mandibular ossifying epulis. A central hemimandibulectomy was performed, and the mandibular defect was stabilized by the use of a free cortical ulnar autograft and rigid internal fixation. The dog had normal mastication and left forelimb function two weeks after surgery. Radiographic evaluation of the surgical site at three and 10 months after surgery showed normal bony healing. PMID- 10667411 TI - Contrast radiographic findings in canine bacterial discospondylitis: a multicenter, retrospective study of 27 cases. AB - A multicenter, retrospective study was undertaken to evaluate contrast radiographic findings in canine bacterial discospondylitis. Records and myelograms or epidurograms of 27 patients were obtained from five colleges of veterinary medicine. Fifteen cases (56%) were evaluated as having some degree of spinal cord compression. The majority (73.3%) of the cases had only soft tissue as the compressive mass. The median compression for all cases was 5% of the vertebral canal. No difference was noted for compression based on anatomical site (i.e., cervical versus thoracolumbar versus lumbosacral). No significant correlation between degree of lesion compression and clinical outcome was noted, but there was a trend toward increased mortality with greater compression. There was no correlation between the ambulatory status and the ultimate outcome. Three of the 15 (20%) cases showed vertebral subluxation. Results of this study indicate that static spinal cord compression is not a significant component of the neurological dysfunction associated with bacterial discospondylitis. Identification of vertebral subluxation in some patients may indicate a dynamic lesion that should be evaluated with stress radiography. PMID- 10667412 TI - How should atypical prostate needle biopsies be reported? Controversies regarding the term "ASAP". PMID- 10667413 TI - ASAP. Atypical small acinar proliferations. PMID- 10667414 TI - ASAP. Atypical small acinar proliferations. PMID- 10667415 TI - ASAP is a valid diagnosis. Atypical small acinar proliferation. PMID- 10667416 TI - The Mi15 monoclonal antibody (anti-syndecan-1) is a reliable marker for quantifying plasma cells in paraffin-embedded bone marrow biopsy specimens. AB - Plasmocyte selective monoclonal antibodies (MAb) recognizing syndecan-1 have recently been described. They belong to a new cluster, CD138. Using the MAb MI15, we investigated the expression of syndecan-1 in routinely paraffin-embedded tissues. Nontumoral lymph nodes (25 cases) and bone marrow biopsy specimens (63 cases) showed strong membrane staining of plasma cells only, allowing accurate analysis of the nuclear structure. The MI15 positivity correlated with kappa and lambda light chain expression in the cytoplasm. The percentages of plasma cells calculated in bone marrow biopsy specimens after MI15 staining were, respectively, 2.1% (range, 1% to 4%) in normal bone marrows, 8.5% (range, 5 to 17) in reactive plasmocytosis, and 4.66% in monoclonal gammapathy of undetermined significance (MGUS) patients (range, 1 to 13), in the same range but slightly higher than those obtained on smears or on hematoxylin and eosin (H&E)-stained sections. In multiple myeloma (40 cases), all plasma cell types were marked, and Mi15 MAb gave additional information in 8 of 40 (20%) patients. In lymph nodes, Mi15 MAb reacted with Reed-Sternberg cells of classical Hodgkin's disease in 23 of 31 cases (74%) with variable intensity. In contrast, nodular lymphocyte predominance Hodgkin's disease (10 cases), most B cell lymphomas (88 of 107 cases) and all T cell lymphomas (30 cases) were negative. In B cell lymphomas, plasmocytomas (8 cases), plasmocytic lymphomas (2 cases), and 5 of 13 cases of immunoblastic lymphoma with plasmocytoid differentiation were stained. In lymphoplasmocytoid lymphomas (4 lymph nodes and 20 bone marrow biopsy specimens), only mature plasma cells were positive. Moreover, a wide distribution of syndecan 1 was observed in normal and tumoral epithelial tissues. Finally, Mi15 MAb appears to be a reliable marker for identifying and quantifying normal and tumoral plasma cells in paraffin-embedded bone marrow and lymph node samples. PMID- 10667417 TI - Cell damage and proliferation in human gastric mucosa infected by Helicobacter pylori--a comparison before and after H pylori eradication in non-atrophic gastritis. AB - Helicobacter pylori (HP) is believed to be involved in the transition from normal gastric mucosa to atrophic gastritis and intestinal metaplasia. Infection with the organism is one of the risk factors for development of intestinal-type gastric adenocarcinoma, possibly through altered cell turnover. Medical eradication of HP is widely performed for the treatment of peptic ulcers and other upper gastrointestinal disorders. Eradication of HP may affect altered cell turnover of the gastric mucosa caused by the infection, but there are few reports comparing sterilized mucosa with HP-infected and non-infected mucosa. In this study, we examined cell damage using terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL), in situ nick translation (ISNT), and cell proliferation by Ki 67 immunohistochemistry staining in gastric mucosa before and after HP eradication and in non-infected gastric mucosa. We then compared these findings using endoscopic gastric biopsy specimens. Labeling indices of TUNEL (2.46 +/- 1.22), ISNT (1.13 +/- 0.42), and Ki67 (21.8 +/- 6.14) in tissue from which HP had been eradicated were significantly lower than those of HP-infected mucosa (6.36 +/- 2.26, 4.00 +/- 1.62, 45.8 +/- 5.35, for TUNEL, ISNT, and Ki67, respectively). There were no significant differences between formerly infected and non-infected mucosa (TUNEL: 2.26 +/- 0.69, ISNT: 1.29 +/- 0.63, Ki67: 23.5 +/- 8.20). These results indicate that medical HP eradication results in decreased cell proliferation and damage, restoring the condition seen in non-infected mucosa. Thus, HP eradication may be effective, not only in the treatment of gastric ulcers or gastric symptoms, but also in the prevention of gastric carcinoma. PMID- 10667418 TI - A comparative study of fibrous dysplasia and osteofibrous dysplasia with regard to expressions of c-fos and c-jun products and bone matrix proteins: a clinicopathologic review and immunohistochemical study of c-fos, c-jun, type I collagen, osteonectin, osteopontin, and osteocalcin. AB - Fibrous dysplasia and osteofibrous dysplasia are both benign fibro-osseous lesions of the bone and are generally seen during childhood or adolescence. Histologically, the features of these bone lesions sometimes look quite similar, but their precise nature remains controversial. We retrospectively studied clinicopathologic findings in 62 cases of fibrous dysplasia and 20 cases of osteofibrous dysplasia with regard to their anatomic location and histological appearance. From among these cases, the immunohistochemical expressions of c-fos and c-jun proto-oncogene products and bone matrix proteins of type I collagen, osteonectin, osteopontin, and osteocalcin were evaluated in 20 typical fibrous dysplasias and 17 osteofibrous dysplasias using paraffin sections, and these expressions were then assessed semiquantitatively. Microscopically, fibrous dysplasia showed various secondary changes, such as hyalinization, hemorrhage, xanthomatous reaction, and cystic change in 22 of the 62 cases (35%). This was a higher incidence than in osteofibrous dysplasia, in which only 2 of the 20 cases (10%) showed such changes. In the elderly fibrous dysplasia cases, the cellularity of fibroblast-like cells was rather low, and those cases were hyalinized. Almost all of the cases of fibrous dysplasia and osteofibrous dysplasia showed positive expressions of c-fos and c-jun products. The expressions of type I collagen and osteopontin showed no difference between fibrous dysplasia and osteofibrous dysplasia. Immunoreactivity for osteonectin in bone matrix was detected in only 1 case of fibrous dysplasia (1 of 20), whereas it was recognized in 14 of the 17 cases of osteofibrous dysplasia. Furthermore, the immunoreactivity for osteocalcin in bone matrix and fibroblast-like cells was higher in fibrous dysplasia than it was in osteofibrous dysplasia, semiquantitatively. Our immunohistochemical results regarding osteonectin and osteocalcin suggest that the bone matrix of fibrous dysplasia is somewhat more mature than that of osteofibrous dysplasia, and that the fibroblast-like cells in fibrous dysplasia share some phenotypic features with osteoprogenitor cells of normal osteogenic tissues. Fibrous dysplasia and osteofibrous dysplasia share some similar histological features, including c-fos and c-jun expressions, although different clinicohistologic features and immunohistochemical expressions of osteonectin and osteocalcin were observed. These features suggest that the mechanisms behind the development of fibrous dysplasia and osteofibrous dysplasia are similar, but this is not necessarily indicative of a closer relationship between the 2 diseases. PMID- 10667419 TI - Decreased expression of protectin (CD59) in gut epithelium in ulcerative colitis and Crohn's disease. AB - Without adequate protection, the cells of the human body would be susceptible to destruction by the complement system. The main defense against complement lysis is a molecule called protectin (CD59) that is widely distributed in human tissues. Because the complement system has been suggested to be involved in the pathogenesis of inflammatory bowel diseases, we examined the expression of protectin in the colonic epithelium of patients with ulcerative colitis or Crohn's disease and controls. Colorectal specimens from 6 patients with ulcerative colitis, 8 patients with Crohn's disease, and 4 controls were obtained from surgical resections. Frozen sections of the specimens were immunostained for protectin using the Bric 229 monoclonal antibody. The expression of protectin was found to be decreased in the epithelium of patients with ulcerative colitis. In patients with Crohn's disease, the epithelial expression of protectin was decreased in diseased areas of gut while the expression did not significantly differ from that in controls in macroscopically normal areas. There was no difference in the expression of protectin on vascular endothelium, mononuclear cells, or smooth muscle. The reduction in epithelial expression of protectin in patients with ulcerative colitis or Crohn's disease may render epithelial cells vulnerable to complement lysis and lead to the destruction of gut epithelium as seen typically in these diseases. PMID- 10667420 TI - Interobserver variability in application of the revised Sydney classification for gastritis. AB - The Sydney classification for gastritis provides guidelines for histological grading of gastric biopsies. In an ongoing study of gastric preneoplastic lesions in Chiapas, Mexico, 7 biopsies from 150 patients (4 from the antrum and 3 from the body) were obtained during endoscopy and studied histologically. The first 74 endoscopy specimens were read independently by 2 general surgical pathologists. We assessed diagnostic concordance using kappa statistics. The 2 pathologists then jointly reviewed biopsies about which they had disagreed to reach a final diagnosis. A second group of 76 endoscopies was subsequently evaluated independently by the 2 pathologists, and concordance was again assessed. In the first group of biopsies, we found low concordance rates (Heliobacter pylori 0.59, acute inflammation 0.22, intestinal metaplasia 0.60, and atrophy 0.04). In the second group, of independently reviewed cases, there was better concordance (H pylori 0.77, acute inflammation 0.50, intestinal metaplasia 0.70, and atrophy 0.64). We presumed that use of the Sydney classification would result in minimal interpretational differences achieving ideal kappas greater than 0.80. Because pathology results are based on subjective interpretation of this classification, complete diagnostic agreement is practically impossible. Concordance by general surgical pathologists after joint review of cases was similar to that obtained by gastrointestinal pathologists. PMID- 10667421 TI - Molecular abnormalities associated with secretory carcinomas of the breast. AB - Secretory carcinomas (SCAs) represent a unique histological variant of invasive breast carcinomas, occurring predominantly in patients younger than 30 years of age. Data from limited series have shown SCAs to have a favorable prognosis in patients younger than 20 years of age, whereas the clinical course tends to parallel the more common in filtrating ductal carcinomas (IDCs) in patients older than 20 years. There are no reports on the molecular abnormalities associated with this unusual tumor. Microdissected archival formalin-fixed tissue from 10 SCAs collected from 2 institutions were used to determine the frequencies of allelic loss at 13 chromosomal regions with 19 microsatellite markers, using multiplex polymerase chain reaction (PCR)-based techniques. The results of loss of heterozygosity (LOH) and microsatellite alterations (MAs) analyses were compared with 20 cases of IDCs. P53 gene mutation analysis was also performed on the 10 SCAs using single-strand conformation polymorphism (SSCP) analysis, followed by sequencing of abnormal bands. LOH at multiple regions of chromosome 3p were the most common abnormality in both SCAs (55%) and IDCs (50%), followed by LOH at 17q21 (BRCA1 locus), 13q14 (retinoblastoma gene locus), and 8p21-23. No significant differences were seen in the frequencies of LOH at any chromosomal region except for 17p13 (p53 gene locus), where allelic losses were absent in SCAs, but evident in 46% of IDCs (P < .05). The 2 histological entities were similar in the fractional regional loss (FRL) index (0.26 v 0.24), fractional allelic loss (FAL) index (0.23 v 0.27), as well as in the frequency of MAs (0.015 v 0.005), P > .05. P53 gene missense mutation (G:C::A:T) was detected in 1 of 10(10%) SCAs. Based on the considerable similarities in the molecular abnormalities associated with both tumors, the formation of secondary lumina in both the in situ and the invasive components, as well as suggestions from limited series that the clinical behavior in adult patients parallels that of IDCs, SCA most likely reflects a secretory variant of IDCs. PMID- 10667422 TI - Crystal-storing histiocytosis: a disorder occurring in plasmacytic tumors expressing immunoglobulin kappa light chain. AB - Intracellular immunoglobulin crystal formation within plasma cells is an uncommon finding in multiple myeloma and other lymphoplasmacytic tumors. We present 12 cases of plasmacytic tumors with prominent crystal formation, including myeloma (5 cases), lymphoplasmacytic lymphoma (6 cases), and a nonneoplastic plasma cell proliferation. In all cases, crystal formation was associated with the proliferation of variable numbers of histiocytes containing similar inclusions. These cases showed a variety of appearances, sometimes obscuring the underlying plasma cell tumor and raising the differential diagnosis of a storage disorder, hemophagocytosis, or a mesenchymal lesion. In cases of lymphoplasmacytic lymphoma, patients typically presented with marked paraproteinemia and symptoms of hyperviscosity. Crystal-storing histiocytosis was not associated with other immunoglobulin deposition disorders, including amyloidosis, Mott cell tumors, or kappa-light chain deposition. In our cases and those previously reported, we found an overwhelming association of crystal-storing histiocytosis (CSH) with tumors expressing immunoglobulin kappa light chain with no consistent association with a particular heavy chain. These results suggest that CSH results from the ingestion of crystals produced by plasma cell tumors that either overproduce kappa light chain or express a structurally aberrant molecule. CSH persists in the marrow and other sites throughout the course of the disease and in our series was not highly associated with development of the adult Fanconi syndrome or rapid clinical deterioration. PMID- 10667423 TI - Colonic aberrant crypts may originate from impaired fissioning: relevance to increased risk of neoplasia. AB - Colonic aberrant crypt foci (ACF) can be identified on the unembedded mucosal surface as clusters of abnormal crypts with enlarged, surface opening. Because dysplasia is frequent, and may be a precursor of carcinoma, epithelial changes have been well studied. However, the basis for the distinctive changes in crypt architecture remain unclear. We hypothesized that some of the architectural alterations of aberrant crypts may reflect impaired fissioning during normal crypt duplication cycles. Fissioning begins at the crypt base. Using morphometric and immunocytochemical approaches, we examined 55 human ACF, both dysplastic and nondysplastic, for their architectural features. Non-ACF mucosa was compared. Microscopically, all lesions contained crypts that were attached, paired, dilated, and angulated. In 3 dimensions, these features related to multiple, individual complexes of connected crypts, referred to as connected crypt structures (CCSs). CCSs terminated in enlarged surface openings (2 to 5 x normal) which are morphometrically equivalent to the macroscopic aberrant crypts (P > .1). These openings trap marker dye. Support for an origin of CCSs in impaired basal fissioning is 3-fold. Crypt profiles in ACF are twice as frequent in basal mucosa as superficially (P < .001); in normal mucosa, the ratio is 1. In a CCS with vertically connected, co-planar crypts, the upper parent crypt diameter was the sum of diameters of inferiorly attached daughter crypts (P > .1). Proliferating cell marker, Ki-67, is not expressed at attachment points. In non ACF mucosa, isolated CCSs consistently occur at foci of mechanical crypt distortion such as mucosal folds. We conclude that a CCS is a fundamental component of ACF of all histotypes. Impairment of normal crypt fissioning is probably a major factor in the histogenesis of CCSs, which often occurs in settings of mechanical distortion of the mucosa. The pathological significance of this process may be in the formation of enlarged crypt openings. The latter could trap dietary carcinogens as they trap dye, and thereby predispose the CCS to dysplasia. PMID- 10667424 TI - Invasive micropapillary carcinoma of the breast: a prognostic study. AB - Invasive micropapillary carcinoma (IMC) of the breast is a rare variant of infiltrating ductal carcinoma that has been associated with an extremely high incidence of lymph node metastases. Follow-up studies on patients with pure IMC breast cancer histology have been limited by low patient numbers, short duration of follow-up, and a lack of multivariate analyses. Using invasive breast cancers from 1,287 patients (median follow-up, 13.8 years), histological review showed 21 cases (1.7%) with pure IMC histology. Pure IMC histology was associated with high grade histology (P = .04), metastases to regional lymph nodes (P < .001), a high mitotic index (P = .02), and erbB-2 immunopositivity (P = .007). Univariate analyses showed a strong association between IMC histology and shortened survival (disease-free survival [DFS], P = .0052; median, 44 months for IMC and 63 months for non-IMC; disease-specific survival [DSS], P = .014; medians, 71 and 78 for IMC and non-IMC, respectively) only in an analysis of all patients. Because only 1 case of node-negative IMC histology was available, univariate analysis of IMC histology was performed only on node-positive patients without significance. Multivariate analyses comparing IMC histology with either node-positive or all other breast cancers failed to show independent prognostic significance. In summary, breast cancer patients with pure IMC histology showed survival rates similar to those of other patients with equivalent numbers of lymph node metastases. PMID- 10667425 TI - Extrathoracic solitary fibrous tumors: their histological variability and potentially aggressive behavior. AB - The histological variability of solitary fibrous tumors may contribute to the difficulty in diagnosing these neoplasms, especially when they arise in extrathoracic sites. Like intrathoracic lesions, the behavior of extrathoracic solitary fibrous tumors is currently unpredictable because these types of tumor have only recently been recognized. This study therefore was undertaken to examine the clinical behavior and histological, immunohistochemical, and ultrastructural features of 24 extrathoracic solitary fibrous tumors with long term follow-up. The patients comprised 10 men and 14 women, between 30 and 85 years of age (mean, 51 years). Ten tumors were located in the retroperitoneum or pelvis, 5 in the trunk, 4 in the extremities, 2 in the orbital region, and 1 each in the kidney, uterine cervix, and meninges. All of the tumors showed a classic morphological appearance, diffuse and strong immunoreactivity for both vimentin and CD34, and variable reactivity for bcl-2. All 7 cases examined ultrastructurally contained fibroblasts and myofibroblasts. Six tumors contained multinucleated giant cells, and in 4 cases these lined pseudovascular spaces with mononuclear cells, thus resembling giant cell angiofibroma and giant cell fibroblastoma. Other potentially similar spindle cell neoplasms mixed with adipose tissue, such as dendritic fibromyxolipoma, lipomatous hemangiopericytoma, cellular angiofibroma, and spindle cell lipoma, were considered in the differential diagnosis. One tumor displayed atypical histological features in the form of increased cellularity and nuclear pleomorphism, but this patient has remained free of disease for 14 years. Another 2 patients developed local recurrences at 6 months and 5 years, and a further patient developed pulmonary metastases that were diagnosed after 7 years. These tumors lacked any atypical histological features in the primary lesions. No patient has so far died of the disease. In conclusion, most extrathoracic solitary fibrous tumors appear to pursue a benign course, although, because some have the potential to recur or metastasize, careful long-term follow-up is necessary for all patients. PMID- 10667426 TI - Demonstrability of the glycoprotein A-80 in postradiation prostatic carcinoma. AB - Radiation therapy results in significant morphological changes in prostatic carcinoma, including decreased cancer size, acinar shrinkage and distortion, cytoplasmic vacuolization, and nuclear pyknosis. Benign acini usually display enlarged, atypical cells with hyperchromatic nuclei. These changes confound the evaluation of limited postradiation samples. The glycoprotein A-80 is known to be upregulated in prostatic intraepithelial neoplasia (PIN) and prostatic carcinoma. In this study, we assessed the expression of A-80 in radiation-treated prostatic carcinoma. Paraffin sections from 64 postradiation prostatic carcinomas obtained at salvage prostatectomy were immunostained with a monoclonal antibody to A-80; selected sections were doubly immunostained with antibodies to A-80 and various cytokeratin polypeptides. All cases showed readily detectable and often intense staining in the cytoplasm of cancer cells and in intraluminal material of malignant acini. The extent and intensity of the reactions were independent of cancer size and grade. Strong reactions were seen in preserved and distorted acini, clear cell areas, single cancer cells, and in colloid pools with few or no recognizable cancer cells. PIN was present in 34 cases (53%), of which 27 (79%) stained strongly for A-80; atrophic and hyperplastic acini generally did not stain, irrespective of the degree of cellular atypia. The A-80 glycoprotein appears remarkably durable and is readily demonstrable in postradiation prostatic carcinoma despite profound architectural and cytologic changes. This characteristic may prove useful in evaluating small samples for confirmation of diagnosis and determination of extent of residual or recurrent prostatic carcinoma after radiation therapy. PMID- 10667427 TI - Living related liver transplantation: histopathologic analysis of graft dysfunction in 304 patients. AB - Between June 1990 and August 1997, 304 mainly pediatric patients underwent a total of 311 orthotopic living related liver transplantations (LRLTs) under tacrolimus immunosuppression at Kyoto University Hospital. Congenital biliary atresia was the most common underlying disease. The donor was a parent, and the left lateral segments were used as grafts in most cases. The average number of loci of HLA-A, -B, and -DR mismatches between the donor and the recipient were 2.1. Forty-three transplants were ABO-incompatible. Liver histology at the time of abnormal liver function after transplantation was analyzed. Preservation injury was rare and mild. Acute cellular rejection (ACR) occurred in 36% of transplants during the first 6 months. Average rejection activity index (the Banff schema) was 4.2 and severe rejection was rarely seen. The number of mismatching HLA loci and immunosuppression regimens affected the incidence of ACR. Chronic rejection (CR) occurred in 2% of transplants. Concerning humoral rejection, no hyperacute rejection was seen. However, hepatic artery thrombosis (delayed hyperacute rejection) was seen in an ABO-incompatible transplant. Acute hepatitis, including those related to cytomegalovirus and Epstein-Barr virus, occurred in 17% of transplants. Chronic hepatitis, including hepatitis B and C, developed in 3%. Acute or chronic cholangitis occurred in 16%, and a significantly higher incidence of cholangitis was found in ABO-incompatible transplants. Posttransplantation lymphoproliferative disease developed in 2%. In LRLT, milder preservation injury and less frequent ACR and CR were suggested, probably because of the short cold-ischemia time and the advantages of HLA histocompatibility, respectively. PMID- 10667428 TI - Significance of acid-mucin-positive nongoblet columnar cells in the distal esophagus and gastroesophageal junction. AB - Acidic mucin-positive nongoblet columnar cells (NGCC) have recently been observed in the surface epithelium of the gastroesophageal junction (GEJ) and distal esophagus in resections from patients with traditional long segment (>3 cm) Barrett's esophagus (BE). However, the significance of finding acidic mucin positive NGCC in the surface epithelium of biopsy specimens from the distal esophagus/GEJ region in the absence of goblet cells (GC) remains unknown. Therefore, to determine the significance of mucin histochemical changes in the distal esophagus/GEJ region, we analyzed and compared the types, prevalence, and distribution of neutral and acidic mucins in biopsy specimens obtained from 2 groups of patients: those with (32 patients) and those without (107 patients) GC identified in this area. Various mucin histochemical stains (PAS-Ab pH 2.5, HID Ab pH 2.5, PB/KOH/PAS) were used to identify neutral mucins, acidic mucins (sialomucins and sulphomucins), and o-acetylated sialomucins. The results were compared between the 2 patient groups and correlated with the clinical, endoscopic, and pathologic features. Compared with patients without GC, patients with GC had a significantly higher male/female ratio and a higher proportion of patients with greater than 3 cm of columnar epithelium within the esophagus. Acidic mucin (sialomucin and sulphomucin)-positive NGCC in the surface, foveolar, and glandular epithelium did not show any correlation with any of the clinical, endoscopic, or pathologic features, such as esophagitis, carditis, antritis, Helicobacter pylori infection, or length of columnar epithelium in the distal esophagus. However, acidic mucin-positive NGCC correlated strongly with the presence of GC (P < .001). For example, sialomucin-positive NGCC were present in 28 of 32 (88%) patients with GC compared with 31 of 107 (29%) patients without GC (P < .001). Similarly, sulphomucin-positive NGCC were present in 20 of 32 (62%) patients with GC, compared with 11 of 107 (10%) patients without GC (P < .001). Of the non-GC cases, all biopsy specimens that stained positively for sulphomucin in surface NGCC (11 specimens), except one, showed staining restricted to the surface of multilayered epithelium, a distinctive type of epithelium that shows morphological, ultrastructural, and cytochemical features of both squamous and columnar epithelium. Sialomucin positivity in surface NGCC from the distal esophagus/GEJ region is a sensitive (sensitivity 88%), but nonspecific (specificity 71%), indicator of GC metaplasia. In contrast, sulphomucin expression in NGCC from the same anatomic area is a less sensitive (sensitivity 62%), but more specific (specificity = 90%) marker for the presence of metaplastic epithelium, of either the GC or the multilayered epithelial cell type and thus may represent an early or incomplete form of intestinal metaplasia. PMID- 10667429 TI - Evidence of SV40 infections in hospitalized children. AB - Simian virus 40 (SV40) is known to have contaminated poliovirus vaccines used between 1955 and 1963. Accumulating reports have described the presence of SV40 DNA in human tumors and normal tissues, although the significance of human infections by SV40 is unknown. We investigated whether unselected hospitalized children had evidence of SV40 infections and whether any clinical correlations were apparent. Serum samples were examined for SV40 neutralizing antibody using a specific plaque reduction test; of 337 samples tested, 20 (5.9%) had antibody to SV40. Seropositivity increased with age and was significantly associated with kidney transplants (6 of 15 [40%] positive, P < .001). Many of the antibody positive patients had impaired immune systems. Molecular assays (polymerase chain reaction and DNA sequence analysis) on archival tissue specimens confirmed the presence of SV40 DNA in 4 of the antibody-positive patients. This study, using 2 independent assays, shows the presence of SV40 infections in children born after 1980. We conclude that SV40 causes natural infections in humans. PMID- 10667430 TI - Persistence of high-grade prostatic intra-epithelial neoplasia under combined androgen blockade therapy. AB - The presence and morphology of high-grade prostatic intraepithelial neoplasia (H PIN) was blindly evaluated in 40 totally embedded radical prostatectomy specimens of patients with prostate cancer randomized to either a 3 (n = 18) or 6 months (n = 22) combined androgen blockade regimen before surgery. In 5 cases, neo-adjuvant therapy was abrogated some time before surgery. In the remaining cases, foci of H PIN were identified in 72% and 59% of prostates from patients treated for 3 and 6 months, respectively. Cellular features used to distinguish H-PIN from normal glands were increased nuclear size, nuclear crowding, anisonucleosis, and disordered nuclear arrangement. In some cases, density of cytoplasm was an additional feature. Unfortunately, the molecular marker erbB2 proved unhelpful for identification of H-PIN. The median number of prostatic glands involved by H PIN was 19 +/- 21 (SD) glands in 3 months treated prostatectomies (n = 18) and 7 +/- 12 (SD) glands in 6 months treated prostatectomies (n = 17), a nonsignificant difference (P = .17). H-PIN was localized within areas of residual carcinoma in 62% and 20%, respectively of prostatectomies of patients treated for 3 and 6 months, respectively. Architectural patterns of H-PIN differed at 3 and 6 months of endocrine pretreatment: The predominant tufted pattern at 3 months was replaced by flat H-PIN at 6 months. The continued expression of androgen receptors and the cell cycle marker MIB-1 in persistent H-PIN suggests that recovery of androgen levels after cessation of androgen blockade therapy will lead to its further expansion. PMID- 10667431 TI - LOH at the sites of the DCC, APC, and TP53 tumor suppressor genes occurs in Barrett's metaplasia and dysplasia adjacent to adenocarcinoma of the esophagus. AB - Barrett's esophagus carries a 30- to 100-fold increased risk of adenocarcinoma, which is thought to develop via a metaplasia-dysplasia-carcinoma progression. A common genetic abnormality detected in Barrett's adenocarcinoma is loss of heterozygosity (LOH) at the sites of known or putative tumor suppressor genes, of which there are at least 9 associated with esophageal adenocarcinoma. The aim of this study was to identify at which histological stage of carcinogenesis LOH at these sites occur. Microdissection of multiple paraffin-embedded tissue blocks from 17 esophagogastrectomy specimens of adenocarcinoma arising in Barrett's esophagus yielded areas of metaplasia, low-, intermediate- and high-grade dysplasia, and carcinoma. LOH analysis of microdissected tissues was performed using a double polymerase chain reaction technique with 11 microsatellite primers shown previously to have LOH in at least 30% of esophageal adenocarcinomas. Identical LOH was detected in premalignant and malignant tissues in 4 of 17 patients, and was located at 5q21-q22 (D5S346 primer), 17p11.1-p12 (TCF2 primer), 17p13.1 (TP53 primer), 18q21.1 (detected in colon cancer tumor suppressor gene [DCC] primer), and 18q23-qter (D18S70 primer). These results suggest that LOH at the sites of the DCC, adenomatous polyposis coli (APC), and TP53 tumor suppressor genes occur before the development of adenocarcinoma in Barrett's esophagus, and so merit further study as potential biomarkers of neoplastic progression in patients with Barrett's esophagus undergoing endoscopic and histological surveillance. PMID- 10667432 TI - Myxoid liposarcoma with transition to round-cell lesion-cell cycle regulator genes and telomerase activity characterizing tumor progression: a case report. AB - A mixed myxoid/round cell liposarcoma was macrodissected in its 2 histologic components and investigated for genetic differences between its low-grade myxoid and the high-grade round-cell region. For both, we failed to detect p53 gene mutations, loss of heterozygosity at the p53 or Rb genes, and p53 protein expression. The round-cell component showed a high telomerase activity, and an elevated c-myc mRNA and protein expression. The myxoid component was characterized by a lack of telomerase activity and low c-myc mRNA expression, and immunohistochemistry failed to detect the c-myc protein. There was a higher Mib-1 proliferation index in the round-cell portion. The same specific translocation t(12;16) and the fusion transcript type II in both components confirmed the close relationship between myxoid and round-cell liposarcomas. Telomerase activity and increased c-myc expression seem to be helpful molecular markers for characterizing tumor progression in myxoid liposarcoma. PMID- 10667433 TI - A case of infantile rhabdomyofibrosarcoma with immunohistochemical, electronmicroscopical, and genetic analyses. AB - A case of infantile rhabdomyofibrosarcoma arising on the buttocks of a 15-month old boy is reported with histological, immunohistochemical, electronmicroscopical, and cytogenetic findings. Histological examination showed a proliferation of spindle-shaped cells in a fasciculated pattern, with occasional rounded rhabdomyoblastic cells with abundant eosinophilic cytoplasm. Immunohistochemically, the tumor cells expressed desmin and MyoD1 but were only weakly positive for myoglobin. No clear rhabdomyoblastic features were observed by electronmicroscopic examination. Chromosome analysis showed a clone of 46, XY, der(2)t(2;11)(q37;q13), different from any karyotypic abnormality in the original report of this neoplasm. Loss of heterozygosity at 11p15.5, the most frequent genetic alteration in embryonal rhabdomyosarcoma, was not detected. The low degree of striated muscle differentiation and tumor localization supported the diagnosis of infantile rhabdomyofibrosarcoma rather than spindle-cell rhabdomyosarcoma in this case. The present case has been uneventful as of 25 months after surgery. The rather long recurrence-free period, which has not been reported in previous cases, may be attributable to chemotherapy-induced rhabdoid differentiation of the tumor cells. PMID- 10667434 TI - Atypical spitz nevus/tumor. PMID- 10667435 TI - Mucoid arteriosclerotic and nerve degenerative lesions in esophageal carcinoma. PMID- 10667436 TI - Enucleation. AB - The three most common indications for enucleation are intraocular malignancy, trauma, and a blind, painful eye. Recommending enucleation is one of the most difficult therapeutic decisions in ophthalmology. In some cases of malignancy, cryotherapy, laser photocoagulation, diathermy, chemotherapy, and radiation therapy may be viable alternatives to surgery. When surgery is chosen, evisceration or exenteration may be alternatives to enucleation. Once the decision is made to perform enucleation or evisceration, the surgeon must choose from several types of implants and wrapping materials. These devices can be synthetic, autologous, or eye-banked tissues. With certain implants, the surgeon must decide when and if to drill for subsequent peg placement. In this review, the authors discuss choices, techniques, complications, and patient consent and follow-up before, during, and after enucleation. Controversies and results of the Controlled Ocular Melanoma Study are summarized. PMID- 10667437 TI - Anatomy of the orbital apex and cavernous sinus on high-resolution magnetic resonance images. AB - Diseases of the orbital apex and cavernous sinus usually present with involvement of multiple cranial nerves, corresponding to the complex anatomy of the region. In nontraumatic disorders, magnetic resonance imaging is the diagnostic modality of choice. However, its capabilities can be fully used only with thorough knowledge of the complicated topographic relationships in this region. This article describes the imaging anatomy of the cranio-orbital junction and adjacent subarachnoid spaces. High-resolution magnetic resonance images of normal subjects are presented, and the results are compared with findings reported in the literature. The following anatomic structures can be visualized on high resolution magnetic resonance images: extraocular muscles and corresponding connective tissue, major orbital and cerebral arteries, ophthalmic veins, cavernous sinus, and all sensory and motor cranial nerves of the eye along their intraorbital and intracranial course. PMID- 10667438 TI - An unusual homonymous visual field defect. AB - A 75-year-old man suddenly became aware of an inferior right homonymous visual field defect. Although static perimetry suggested a lesion of the left lateral geniculate nucleus, kinetic perimetry indicated that the presumed homonymous horizontal sectoranopia noted on static perimetry was actually an incomplete homonymous hemianopia with incomplete sparing of the temporal crescent. The location of the lesion was subsequently confirmed by magnetic resonance imaging. This case shows the value of kinetic perimetry in assessing homonymous visual field defects. PMID- 10667439 TI - Chiasmopathy? AB - A 57-year-old man presented with progressive visual loss in both eyes, bitemporal field defect, and a history of poor nutrition, alcohol abuse, and excessive cigar smoking. Magnetic resonance imaging was normal. The visual acuity and field defect improved with supplementation with vitamins and reduction of alcohol and tobacco consumption. A diagnosis of toxic optic neuropathy was made. The authors discuss the differential diagnosis of bitemporal/pseudobitemporal field defects and the diagnosis and treatment of toxic optic neuropathy. PMID- 10667440 TI - Leiomyoma of the ciliary body extending to the anterior chamber: clinicopathologic and ultrasound biomicroscopic correlation. AB - Leiomyoma of the ciliary body is a rare tumor that often causes a diagnostic dilemma. Sclerouvectomy has been found to be beneficial in the management of iris and ciliary body leiomyoma. We treated a case of leiomyoma of the ciliary body presenting as a fleshy mass in the anterior chamber, removed by partial lamellar sclerouvectomy. Ultrasound biomicroscopic and histopathologic features, including light microscopic, immunohistochemistry, and transmission electron microscopic features, were studied and the literature was reviewed. PMID- 10667441 TI - A brief history of macular grids: from Thomas Reid to Edvard Munch and Marc Amsler. AB - Metamorphopsia is a symptom of retinal distortion from intrinsic retinal disease. It has undoubtedly been experienced for millennia, but its clinical significance has been appreciated only in modern times. The Norwegian painter Edvard Munch recognized scotomas and metamorphopsia after suffering an intraocular hemorrhage in his 60th year. Drawings made during this illness show his changing perceptions, and also his attempts to document them with a grid of lines. The Scottish philosopher Thomas Reid may have been the first to write about metamorphopsia. He described distortion of his vision in 1764, after an episode of sungazing, and recognized that the problem was probably of retinal origin. Lines or grids to document metamorphopsia have appeared in ophthalmology textbooks for more than 100 years, but testing for macular degeneration did not become routine until the dissemination of Amsler's grids in the middle of the 20th century. This is in large measure a result of developments in ophthalmology that made therapy for macular disease possible. PMID- 10667442 TI - Measure for measure. AB - Shakespeare's play Measure for Measure colorfully illustrates a concept that is relevant to the testimonies given by the medical expert witness in the courtroom. The expert should not ask the jury to hold the defendant physician to a higher standard than that actually practiced by an acceptable reasonable minority of the defendant's peers under similar circumstances. Two cases are presented. PMID- 10667443 TI - The development of the code of ethics for the American Academy of Ophthalmology. AB - The circumstances leading to and the process of establishing a code of ethics for members of the American Academy of Ophthalmology are recounted by the original chairman of the Ethics Committee. PMID- 10667444 TI - Avoiding misplacement of blame for medical maloccurrence. PMID- 10667445 TI - Managing retained intravitreal lens fragments after cataract surgery. PMID- 10667446 TI - The placebo effect. PMID- 10667447 TI - Hormones and cognition: current concepts and issues in neuropsychology. AB - This article provides an extensive and comprehensive review of the effects of hormones on cognition. Studies detailing specific neurocognitive functions affected by variation in hormone levels across the life span are presented. Dysregulation of hormone levels is considered from models of both normal and diseased functioning. Patterns of cognitive dysfunction are described for a range of syndromes involving the neuroendocrine system, and evidence of specific neurophysiological mechanisms that can account for these findings is outlined. This review includes discussion of treatment outcomes and the permanency of endocrine-related cognitive dysfunction. The authors present a set of guidelines for clinical neuropsychologists to use for assessment of patients with neuroendocrine system dysfunction. Clinical and methodological issues in research and treatment settings are discussed. PMID- 10667448 TI - Quality of life and neurological illness: a review of the literature. AB - This article highlights some of the conceptual and methodological problems associated with quality of life (QoL) measurement in individuals with neurological illness. It is suggested that these problems have contributed to the underdeveloped status of QoL research in neurological settings. Many of the existing QoL measures that have been used, or show potential for use with individuals with neurological illness, are reviewed in terms of their theoretical basis, content, and practicality. A large proportion of these measures fail to meet adequate psychometric standards and/or have rarely been psychometrically tested. The confusion that surrounds adequate psychometric standards is discussed, and the dynamic nature of QoL is highlighted as a factor that requires further attention. Research addressing the discrepancies between proxy and patient ratings of patient QoL is also warranted. More focused research in these areas may contribute to a clearer understanding of how to assess QoL in individuals with neurological illness. PMID- 10667449 TI - Cognitive event-related potentials in neuropsychological assessment. AB - Neuropsychology is a behavioral approach to studying the brain, and an integration of neuropsychology with on-line processing measures of brain function is important for advancing the understanding of brain-behavior relationships. Cognitive event-related potentials (ERPs) are on-line processing measures that are of interest to neuropsychologists because they are linked to familiar neuropsychological test paradigms and because they have reached a degree of standardization sufficient to make them applicable in individual assessment. A selective review of cognitive ERPs is given, focusing on studies of attention in the oddball paradigm and arguing that an adequate assessment of attention is basic in understanding higher order cognitive functions. General principles for using ERP data to supplement and clarify neuropsychological analysis are discussed, and available evidence on dementia and traumatic head injury is reviewed. It is concluded that ERPs are a useful supplement to neuropsychological assessment. Although diagnostic use of ERPs must be guarded because of limited standardization and validation, information-processing analysis with ERPs may aid significantly in interpretation of behavioral data. PMID- 10667450 TI - The search for circadian clock and sleep genes. AB - In recent years, there has been extraordinary progress in elucidating the molecular components of the mammalian circadian clock system. The discovery of circadian clock genes in lower organisms (such as fruit flies and fungi), which show many similarities with clock genes in mammals, together with advances in mouse molecular genetics have led to major new discoveries on the molecular and genetic basis of mammalian circadian rhythms. This article reviews both of these lines of research from an historical perspective and discusses how these lines have merged to provide unique insights into the molecular mechanisms of circadian function. The review also speculates on how the discovery of circadian clock genes may lead directly or indirectly to the discovery of mammalian sleep genes. The determination of the molecular mechanisms via which circadian clock genes (and their protein products) regulate the timing and the need for sleep, and the identification of new genes involved in sleep regulation, may produce new information on the genetic and molecular control of sleep which could ultimately lead to the development of new treatments for sleep disorders. PMID- 10667451 TI - New insights into the mechanism of action of hypnotics. AB - Between 1987 and 1989, the different protein subunits that make up the receptor for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) were identified. These make up the alpha, beta, gamma and delta families, for each of which exist several subtypes. This receptor is the molecular target of modern hypnotic drugs (i.e. benzodiazepines, zopiclone, zolpidem and zaleplon). In the 10 years that have followed this milestone, significant progress has been made in exploring the molecular mechanisms of hypnotic drug action. Receptor subtype specificity of hypnotics has been explained in terms of differential affinity for receptors containing different alpha subunits, which are expressed in different brain regions. Zolpidem and zaleplon bind preferentially to alpha1-containing receptors, whereas benzodiazepines and zopiclone are aspecific. Different sets of subunits are encoded in contiguous 'cassettes' on the genome, and the transcription of each set appears to be regulated coherently. The predominant GABA(A) receptor composition found in the brain is alpha1beta2gamma2, which are all encoded on human chromosome 5. Targeted gene disruption has provided clues to the physiological functions served by GABA(A) receptors containing different subunits. Receptors containing gamma2 appear to have a vital role in maintaining appropriate central inhibition, beta3-containing receptors may also be important determinants of excitability in certain brain regions, whereas a clear role for alpha5-, alpha6- and gamma3-containing receptors has not yet been established by these techniques. Site-directed mutagenesis has indicated that benzodiazepines bind to a cleft on the GABA(A) receptor surface at the interface between the alpha and gamma subunits. Other drugs (flumazenil, zopiclone, zolpidem) also bind to the a subunit, but interact with amino acids in different binding domains to the benzodiazepines. The molecular mechanism of hypnotic dependence has been explored, and seems to involve downregulation of transcription of the normally prevalent alpha1, beta2 and gamma2 subunits, and the reciprocal upregulation of the expression of rarer subunits. Chronic treatment with hypnotic drugs that may have less dependence potential, such as zopiclone and zolpidem, appears to produce more limited change in GABA(A) receptor subunit expression. These ideas will be important both for designing new hypnotic drugs with a better safety/efficacy profile, and for evaluating more appropriate ways of using the drugs available today. PMID- 10667452 TI - Evaluation of severe insomnia in the general population: results of a European multinational survey. AB - The epidemiology of severe insomnia and its effect on quality of life and healthcare consumption was assessed in a survey of the general population of five northern European countries. Applying established consumer sampling techniques, insomnia sufferers were selected from the general population using a questionnaire, conducted by face-to-face interview, and severity of insomnia was ranked (severe, mild/moderate, no sleep complaint) using a specific algorithm. Population samples were matched according to case control methodology for age, gender and geographical region. A second questionnaire gathered information on sleep problems, quality of life (SF-36 scores) and healthcare consumption. The prevalence of severe insomnia ranged from 4% to 22%, was higher in females than in males, but did not increase significantly with age. Patients with severe insomnia had been experiencing sleeping problems for a median of 2-6 years. In all countries, insomnia had a negative impact on quality of life, and the degree of impairment in quality of life was directly related to the severity of insomnia. Individuals with severe insomnia also showed a higher level of healthcare consumption. Despite this, severe insomnia did not appear to feature prominently in the doctor-patient relationship. PMID- 10667453 TI - Brain mechanisms of sleep: contribution of neuroimaging techniques. AB - Functional brain neuroimaging essentially relies on two basic principles: functional segregation and functional interaction. Recent studies of the functional segregation of the human brain during sleep, using positron emission tomography and statistical parametric mapping, indicate that human brain function is organized in a very specific way in each state of vigilance. During slow wave sleep, the most deactivated areas are the upper brainstem, thalamic nuclei and basal forebrain; deactivation of the basal ganglia is also seen. In the cortex, the least active areas are the associative cortices of the frontal and parietal cortex. The anatomical extent of this deactivation remains uncertain. In rapid eye movement (REM) sleep there is significant activation of the dorsal tegmentum of the ponto-mesencephalic region and the thalamic nuclei. Within the cortex, the limbic areas (amygdala, hippocampus, orbito-frontal cortex and anterior cingulate cortex) are activated. In contrast, the associative areas of the frontal and parietal cortices are less active than other parts of the brain. The functional interactions between the amygdala and the temporal cortex during REM sleep differ markedly from those during other states of vigilance, and there is also an inverse relationship between the activities of the primary and secondary visual areas. PMID- 10667454 TI - Evaluation of severe insomnia in the general population--implications for the management of insomnia: focus on results from Ireland. PMID- 10667455 TI - Evaluation of severe insomnia in the general population--implications for the management of insomnia: the German perspective. PMID- 10667456 TI - Evaluation of severe insomnia in the general population--implications for the management of insomnia: focus on results from Belgium. PMID- 10667457 TI - Evaluation of severe insomnia in the general population--implications for the management of insomnia: the UK perspective. PMID- 10667458 TI - Evaluation of severe insomnia in the general population--implications for the management of insomnia: insomnia, quality of life and healthcare consumption in Sweden. PMID- 10667459 TI - New trends in insomnia management. AB - Pharmacological management of insomnia is continually evolving. The introduction of non-benzodiazepine benzodiazepine receptor agonist hypnotics provides an opportunity to understand different patterns of pharmacological activity with mechanistic differences in receptor activity. The impact of insomnia on daytime functioning and long-term health and socioeconomic status has been recognized. Epidemiological studies indicate that insomnia is associated with increased absenteeism and healthcare costs (although the latter appear to be partly attributable to comorbid depression). It will thus be important to determine whether hypnotics, in addition to their effects on sleep, provide other benefits for the patient that are related to these parameters. The successful resolution of this issue will require the adoption of additional outcome measures, such as effects on quality of life and healthcare costs. Recognition that long-term hypnotic use is widespread among insomniacs has prompted proposals for alternative prescribing patterns. Although certain hypnotics appear to be free of tolerance on prolonged use, caution is required in the long-term use of any hypnotic because of the lack of systematic data on chronic efficacy and safety. Therefore, alternative administration schedules (e.g. intermittent and 'as required') are being investigated. These may have very different consequences in terms of abuse liability and patients' perceptions of efficacy, and thus permit a more effective and appropriate use of this drug class. PMID- 10667460 TI - Genetic polymorphism of CYP genes, alone or in combination, as a risk modifier of tobacco-related cancers. AB - Tobacco use is causally associated with cancers of the lung, larynx, mouth, esophagus, kidneys, urinary tract, and possibly, breast. Major classes of carcinogens present in tobacco and tobacco smoke are converted into DNA-reactive metabolites by cytochrome P450 (CYP)-related enzymes, several of which display genetic polymorphism. Individual susceptibility to cancer is likely to be modified by the genotype for enzymes involved in the activation or detoxification of carcinogens in tobacco and repair of DNA damage. We summarize here the results of case-control studies published since 1990 on the effects of genetic variants of CYP1A1, 1A2, 1B1, 2A6, 2D6, 2E1, 2C9, 2C19, 17, and 19 alone or in combination with detoxifying enzymes as modifiers of the risk for tobacco-related cancers. The results of studies on gene-gene interactions and the dependence of smoking related DNA adducts on genotype were also analyzed. Some CYP variants were associated with increased risks for cancers of the lung, esophagus, and head and neck. The risk was often increased in individuals who also had GSTM1 deficiency. For breast cancer in women, a few studies suggested an association with CYPs related to metabolism of tobacco carcinogens and steroidal hormones. The overall effects of common CYP polymorphisms were found to be moderate in terms of penetrance and relative risk, with odds ratios ranging from 2 to 10. Some CYP1A1/GSTM1 0/0 genotype combinations seem to predispose the lung, esophagus, and oral cavity of smokers to an even higher risk for cancer or DNA damage, requiring, however, confirmation. Future strategies in molecular cancer epidemiology for identifying such susceptible individuals are discussed with emphasis on well-designed larger studies. PMID- 10667461 TI - Molecular genetics and epidemiology of the NAT1 and NAT2 acetylation polymorphisms. AB - The focus of this review is the molecular genetics, including consensus NAT1 and NAT2 nomenclature, and cancer epidemiology of the NAT1 and NAT2 acetylation polymorphisms. Two N-acetyltransferase isozymes, NAT1 and NAT2, are polymorphic and catalyze both N-acetylation (usually deactivation) and O-acetylation (usually activation) of aromatic and heterocyclic amine carcinogens. Epidemiological studies suggest that the NAT1 and NAT2 acetylation polymorphisms modify risk of developing urinary bladder, colorectal, breast, head and neck, lung, and possibly prostate cancers. Associations between slow NAT2 acetylator genotypes and urinary bladder cancer and between rapid NAT2 acetylator genotypes and colorectal cancer are the most consistently reported. The individual risks associated with NAT1 and/or NAT2 acetylator genotypes are small, but they increase when considered in conjunction with other susceptibility genes and/or aromatic and heterocyclic amine carcinogen exposures. Because of the relatively high frequency of some NAT1 and NAT2 genotypes in the population, the attributable cancer risk may be high. The effect of NAT1 and NAT2 genotype on cancer risk varies with organ site, probably reflecting tissue-specific expression of NAT1 and NAT2. Ethnic differences exist in NAT1 and NAT2 genotype frequencies that may be a factor in cancer incidence. Large-scale molecular epidemiological studies that investigate the role of NAT1 and NAT2 genotypes and/or phenotypes together with other genetic susceptibility gene polymorphisms and biomarkers of carcinogen exposure are necessary to expand our current understanding of the role of NAT1 and NAT2 acetylation polymorphisms in cancer risk. PMID- 10667462 TI - Single-dose administration of Bowman-Birk inhibitor concentrate in patients with oral leukoplakia. AB - The Bowman-Birk inhibitor (BBI) is a soybean-derived serine protease inhibitor and a potential cancer chemopreventive agent for humans. In this Phase I clinical trial, BBI concentrate was administered as a single oral dose to 24 subjects with oral leukoplakia. Pharmacokinetics of BBI was analyzed, and subjects were monitored clinically for toxic effects. Subjects received between 25 and 800 chymotrypsin inhibitor units (CIU) of the compound in a dose escalation trial. BBI was taken up rapidly, and a metabolic product of BBI was excreted in the urine within 24-48 h. No clinical or laboratory evidence of toxicity was observed in the study. Protease activity was also measured in buccal cells to evaluate usefulness as a biomarker. Single-dose BBI concentrate administered up to 800 CIU was well tolerated and appeared to be nontoxic. Further investigation in Phase II clinical trials is being done. PMID- 10667463 TI - Selenium modulation of cell proliferation and cell cycle biomarkers in normal and premalignant cells of the rat mammary gland. AB - The present study was designed to assess the effect of Se-methylselenocysteine or triphenylselenonium chloride treatment on cell proliferation [bromodeoxyuridine (BrdUrd) labeling] and cell cycle biomarkers [proliferating cell nuclear antigen (PCNA), cyclin D1, and p27/Kip 1] in the intact mammary gland of rats. Immunohistochemical assays of the above end points were carried out in different morphological structures: (a) terminal end bud cells and alveolar cells of a maturing mammary gland undergoing active differentiation; and (b) premalignant mammary intraductal proliferations (IDPs) identified at 6 weeks after carcinogen dosing. Neither compound was found to affect BrdUrd labeling or the expression of cell cycle biomarkers in the normal terminal-end bud cells and alveolar cells. Se methylselenocysteine reduced the total number of IDP lesions by approximately 60%. Interestingly, this was not accompanied by decreases in BrdUrd labeling or the proportion of IDP cells expressing PCNA and cyclin D1. An enhancement in the fraction of p27/Kip 1-positive IDP cells, however, was detected as a result of Se methylselenocysteine treatment. Although triphenylselenonium chloride did not reduce the total number of IDPs, there were more of the smaller-sized lesions and fewer of the larger-sized lesions compared with those found in the control group. Triphenylselenonium chloride also significantly decreased the proportion of IDP cells incorporating the BrdUrd label or expressing PCNA and cyclin D1. The above findings suggest that early transformed cells are sensitive to selenium intervention, whereas normal proliferating cells are not. It is possible that Se methylselenocysteine blocks carcinogenesis by a pathway that may not involve cell growth inhibition as a primary response; in contrast, triphenylselenonium chloride is likely to act by a cytostatic mechanism. The data also imply that selenium efficacy testing in intervention trials is possible with the use of biomarkers, provided that the appropriate biomarkers are matched with the selenium compound of interest and that the pathological characteristics of the cell population to be evaluated are taken into consideration. PMID- 10667464 TI - Breast adipose tissue concentrations of polychlorinated biphenyls and other organochlorines and breast cancer risk. AB - Numerous studies have examined the relationship between organochlorines and breast cancer, but the results are not consistent. In most studies, organochlorines were measured in serum, but levels in breast adipose tissue are higher and represent cumulative internal exposure at the target site for breast cancer. Therefore, a hospital-based case-control study was conducted in Ontario, Canada to evaluate the association between breast cancer risk and breast adipose tissue concentrations of several organochlorines. Women scheduled for excision biopsy of the breast were enrolled and completed a questionnaire. The biopsy tissue of 217 cases and 213 benign controls frequency matched by study site and age in 5-year groups was analyzed for 14 polychlorinated biphenyl (PCB) congeners, total PCBs, and 10 other organochlorines, including p,p'-1,1-dichloro 2,2-bis(p-chlorophenyl)ethylene. Multiple logistic regression was used to assess the magnitude of risk. While adjusting for age, menopausal status, and other factors, odds ratios (ORs) were above 1.0 for almost all organochlorines except five pesticide residues. The ORs were above two in the highest concentration categories of PCB congeners 105 and 118, and the ORs for these PCBs increased linearly across categories (Ps for trend < or =0.01). Differences by menopausal status are noted especially for PCBs 105 and 118, with risks higher among premenopausal women, and for PCBs 170 and 180, with risks higher among postmenopausal women. Clear associations with breast cancer risk were demonstrated in this study for some PCBs measured in breast adipose tissue. PMID- 10667465 TI - Risk of breast cancer according to the status of HER-2/neu oncogene amplification. AB - We examined risk factors for breast cancer after subdividing cases based on the presence of HER-2/neu oncogene amplification in their tumors. Data were from the Carolina Breast Cancer Study, a population-based, case-control study of 577 invasive breast cancer patients, diagnosed during 1993-1996 and ages 20-74 years, and 790 controls frequency-matched on race and age. Information on breast cancer risk factors was obtained from structured personal interviews. About 20% of paraffin-embedded tissues from the breast cancers of cases were identified as positive for HER-2/neu amplification (HER-2/neu+) by differential PCR. Early age at menarche, higher waist:hip ratio, and family history of breast or ovarian cancer were associated with elevated odds ratios (ORs) for both HER-2/neu+ and HER-2/neu- breast cancers. Breastfeeding for at least 1 year was inversely associated with HER-2/neu+ breast cancer [OR, 0.3; 95% confidence interval (CI), 0.1-0.7] more so than HER-2/neu- breast cancer (OR, 0.8; 95% CI, 0.5-1.2). Most of the remaining risk factors had ORs around 1.0 for both HER-2/neu+ and HER 2/neu- breast cancers, although a few exhibited possible associations with one disease subtype in analyses stratified by menopausal status. These study results suggest that most recognized breast cancer risk factors do not operate through HER-2/neu amplification in breast carcinogenesis. Differential effects of long term breastfeeding by HER-2/neu amplification status have been observed in earlier studies and are provocative; however, the direction and magnitude of the associations have not been consistent. PMID- 10667466 TI - GSTT1 and GSTM1 null genotypes and the risk of gastric cancer: a case-control study in a Chinese population. AB - Glutathione S-transferase (GST) enzymes are involved in detoxification of many potentially carcinogenic compounds. The homozygous deletions or null genotypes of GSTT1 (theta class) and GSTM1 (mu class) genes may be associated with an increased risk of cancer. Few studies have evaluated the relationship between GSTT1, GSTM1 and the risk of gastric cancer, as well as the potential interactions between these genetic markers and other risk factors of gastric cancer in the Chinese population. We conducted a case-control study with 143 cases with gastric cancer, 166 chronic gastritis (CG) cases and 433 cancer-free population controls from Yangzhong County, China. The epidemiological data were collected by a standard questionnaire for all of the subjects, and blood samples were obtained from 91 gastric cancer cases, 146 CG cases, and 429 controls. GSTT1 and GSTM1 genotypes were assayed by the PCR method, and Helicobacter pylori infection was measured by the ELISA method. Using logistic regression model in SAS, we assessed the independent effects of GSTT1 and GSTM1 null genotypes on the risk of gastric cancer and their potential interactions with other factors. The prevalence of GSTM1 null genotype was 48% in gastric cancer cases, 60% in CG patients, and 51% in controls. The prevalence of GSTT1 null genotype was 54% in gastric cancer cases, 48% in CG patients, and 46% in controls. After controlling for age, gender, education, pack-years of smoking, alcohol drinking, body mass index, H. pylori infection, and fruit and salt intake, the adjusted odds ratio (OR) for GSTT1 and gastric cancer was 2.50 (95% confidence interval (CI), 1.01 6.22). When gastric cancer cases were compared with CG patients, the adjusted OR for GSTT1 was 2.33 (95% CI, 0.75-7.25). However, GSTT1 null genotype was not associated with the risk of CG when using population controls. No obvious association was found between GSTM1 and the risk of both gastric cancer and CG. Our results suggest that GSTT1 null genotype may be associated with an increased risk of gastric cancer in a Chinese population. PMID- 10667467 TI - A new ELISA kit for measuring urinary 2-hydroxyestrone, 16alpha-hydroxyestrone, and their ratio: reproducibility, validity, and assay performance after freeze thaw cycling and preservation by boric acid. AB - There is considerable controversy regarding the role of estrogen metabolites in breast cancer risk, fueled in part by the development of a rapid ELISA that is suitable for large scale investigations. An earlier version of the ELISA could detect values of the 2-hydroxyestrone (2-OHE1) and 16alpha-hydroxyestrone (16alpha-OHE1) metabolites as low as 2 ng/ml and produce consistent results in premenopausal urines. However, reproducibility was problematic in postmenopausal urines where concentrations of these compounds are much lower. In response to our concern, a new ELISA was developed with a sensitivity of 0.625 ng/ml, which we evaluated using the same pre- and postmenopausal urine samples analyzed in the earlier ELISA. In this report, we present findings on the new kit with regard to reproducibility of the 2-OHE1 and 16alpha-OHE1 measurements, comparability of results with gas chromatography-mass spectroscopy values, and with regard to the stability of the metabolites after repeated freeze-thaw cycles and after preservation by boric acid. For the most part, we found the new ELISA to be reproducible, with assay coefficients of variation ranging from 10 to 20%, and intraclass correlation coefficients (ICCs) ranging from 80 to 95% in both the pre and postmenopausal urines. ELISA results for 16alpha-OHE1 differed from 1 day (i.e., batch) to the next, and the absolute values of the metabolites obtained by the ELISA were consistently lower than but well correlated with those obtained by gas chromatography-mass spectroscopy. Values of the 2-OHE1:16alpha-OHE1 ratio also differed between the methods, but because the range of values was not large, the magnitude of these differences was not as great. For the ratio, the correlation between methods was excellent, and the ICCs were high for both groups of women. After preservation by boric acid, values of the ratio varied according to acid concentration but not in a linear fashion. Ratio values were similar in urine samples exposed to four different freeze-thaw cycle treatments, although values for all treatments were consistently lower in one batch. Because batch-to batch variability was not negligible, it is advisable that matched cases and controls be analyzed in the same batch. Provided this is done, the relatively low assay coefficient of variation and high ICC demonstrate that the new ELISA kit can reliably measure the 2-OHE1:16alpha-OHE1 ratio and detect small case-control differences in large population-based studies, where rapid and relatively easy laboratory methods are critical. PMID- 10667468 TI - Behavioral risk factors among women presenting for genetic testing. AB - Considerable research attention has been given to the impact of genetic testing on psychological outcomes. Participation in genetic testing also may impact on health behaviors that increase the risk of cancer and other chronic diseases. The purpose of this study is to describe behavioral cancer risk factors of women who requested genetic testing for breast and ovarian cancer susceptibility (BRCA1, BRCA2). Before participation in a genetic testing program, 119 women completed a series of questionnaires designed to assess their health behaviors, perception of risk, and depressive symptomatology. Eight percent of participants were current smokers, 27% did not engage in at least moderate exercise, 46% did not regularly protect themselves from the sun, 39% did not consume at least five servings of fruits and vegetables per day, and 9% drank at least one alcoholic beverage per day. Poisson regression analysis revealed that age was the only predictor of behavioral risk profiles, with older women having fewer cancer risk behaviors. These patients who presented for genetic testing generally had better health behaviors than the general population. However, given their possible high-risk status, these patients should consider further improving their preventable cancer risk factors and, in particular, their diet, sun protection, and physical activity levels. Inclusion of behavioral risk factor counseling in the context of the genetic testing process may be an important opportunity to reach this at-risk population. PMID- 10667469 TI - A case-control study of galactose consumption and metabolism in relation to ovarian cancer. AB - Consumption or metabolism of dairy sugar and ovarian cancer have been linked based on evidence that galactose may be toxic to ovarian germ cells and that ovarian cancer is induced in animals by depletion of oocytes. We assessed consumption of dairy products and obtained blood for biochemical and molecular genetic assessment of galactose metabolism in 563 women with newly diagnosed epithelial ovarian cancer and 523 control women selected either by random digit dialing or through lists of residents in eastern Massachusetts and New Hampshire. We observed no significant differences between cases and controls in usual consumption of various types of dairy products or total daily lactose (the principal source of galactose in the diet); nor did we find that RBC activity of either galactose-1-phosphate uridyl transferase (GALT) or galactokinase differed. The mean (and SE) activity of uridine diphospho-galactose 4'-epimerase (in micromoles per hour per gram of hemoglobin) was, however, significantly lower (P < 0.005) in cases compared with controls, 20.32 (0.31) versus 21.64 (0.36). Ovarian cancer cases were also more likely to carry the N314D polymorphism of the GALT gene, generally predisposing to lower GALT activity. The difference was most evident for endometrioid and clear cell types of ovarian cancer, in which 3.9% of cases were found to be homozygous for N314D compared with 0.4% of controls, yielding an odds ratio and 95% confidence interval of 14.17 (2.62-76.60). We conclude that, whereas adult consumption of lactose carries no clear risk for the disease, certain genetic or biochemical features of galactose metabolism may influence disease risk for particular types of ovarian cancer. PMID- 10667470 TI - Cancer risk estimates for family members of a population-based family registry for breast and ovarian cancer. AB - Population-based breast and ovarian cancer family registries can facilitate studies to evaluate genetic and environmental factors in the etiology of these malignancies. The purpose of this study is to describe what is, as far as we know, the first population-based breast and ovarian cancer family registry and to estimate breast and ovarian cancer risk in relatives of breast and ovarian cancer probands. Population-based consecutive incident cases of breast and ovarian cancer were invited to participate in the University of California, Irvine breast and ovarian family registry. In this study, we report data on 1567 breast cancer and 328 ovarian cancer probands. The operational components of this family registry include enrollment of probands, family history interviewing, confidentiality, pathology, verification and review, biospecimen bank, statistical/genetic analysis, and special studies on positional cloning of known genes. All of the components are tracked through the University of California, Irvine Genetic Research Information System. In non-Hispanic-white breast cancer probands, relative risk (RR) of breast cancer in mothers and sisters is significantly elevated [RR = 1.7 and 95% confidence interval (CI) = 1.4-2.0 and RR = 2.8 and 95% CI = 2.3-3.3, respectively]. In families of ovarian cancer probands, mothers are at increased risk of ovarian cancer (RR = 4.6; 95% CI, 2.1 8.7). RR of breast cancer in mothers of Hispanic breast cancer probands is significantly elevated (RR = 4.9; 95% CI, 2.6-8.5). No elevation of breast or ovarian cancer risk was observed among relatives of Asian probands. In general, there is a decrease in RR among mothers and sisters with increase in age of onset of probands. In second-degree relatives and first cousins, the breast cancer hazards ratios increase with increase in the number of affected first-degree relatives and decrease with increase in age at onset of the proband. PMID- 10667471 TI - Diets containing whey proteins or soy protein isolate protect against 7,12 dimethylbenz(a)anthracene-induced mammary tumors in female rats. AB - A study was conducted to determine the protective effects of two common dietary proteins, soy protein isolate (soy) and bovine whey, against chemically induced mammary tumors in female Sprague Dawley rats. Rats were fed AIN-93G diets having casein, soy, or whey as the sole protein source. Rats within the same dietary groups were mated to obtain the F1 and F2 generations. At age 50 days, F1 (experiment A) or F2 (experiment B) female offspring (> or =19 rats/group) were p.o. gavaged with 80 mg/kg 7,12-dimethylbenz(a)anthracene, and mammary glands were evaluated when 100% of the casein-fed group developed at least one palpable tumor. Rats grew well on all three diets, but casein-fed rats gained slightly more body weight than soy- or whey-fed rats (P < 0.05). Vaginal opening occurred 1 day earlier in soy-fed rats than in casein- or whey-fed rats, but no other differences in reproductive and developmental parameters were observed between groups. When 50% of the casein-fed rats had at least one mammary tumor, lower tumor incidences (24-34%) were observed in the soy-fed (P < 0.009) and whey-fed groups (P < 0.001). When 100% of the casein-fed rats had at least one tumor, soy fed rats had a lower tumor incidence (77%) in experiment B (P < 0.002), but not in experiment A (P < 0.12), and there were no differences in tumor multiplicity. Whey-fed rats had lower mammary tumor incidence (54-62%; P < 0.002) and multiplicity (P < 0.007) than casein-fed rats in both experiments. Our results indicate that diets rich in soy reduce the incidence of chemically induced mammary tumors by approximately 20%. Furthermore, whey appears to be at least twice as effective as soy in reducing both tumor incidence and multiplicity. PMID- 10667472 TI - Nonsteroidal anti-inflammatory drugs and risk of digestive cancers at sites other than the large bowel. AB - Regular continuing nonsteroidal anti-inflammatory drug (NSAID) use has been associated with a reduction in risk of large bowel cancer in many studies, including our Case-Control Surveillance Study of medication use and cancer risk. We assessed the relation of NSAID use to the risk of digestive cancers at sites other than the large bowel in this database. Nurse-interviewers administered questionnaires to patients admitted to hospitals in four centers from 1977 to 1998. Cases comprised 1149 patients with cancers of the pancreas (n = 504), stomach (n = 254), esophagus (n = 215), gallbladder (n = 125), or liver (n = 51). Controls were 5952 patients admitted for trauma or acute infection. History of NSAID use was elicited by questions about indications for use. Multiple logistic regression models were used to calculate odds ratios (ORs) for categories of regular NSAID use (at least 4 days/week for at least 3 months) relative to never use. The OR for regular use initiated at least 1 year before admission and continuing into that year was reduced for stomach cancer (OR = 0.3; 95% confidence interval, 0.1-0.6) and was compatible with 1.0 for other sites. The ORs for regular continuing use of at least 5 years duration were < 1.0 for cancers of the stomach, pancreas, esophagus, and gallbladder but were statistically significant only for stomach cancer. These data suggest that regular continuing NSAID use may be associated with reduced risk of stomach cancer. For the other sites, the data are consistent with no effect of NSAID use. PMID- 10667473 TI - Analytical and clinical evaluation of TSH and thyroid hormones by electrochemiluminescent immunoassays. AB - OBJECTIVES: To perform an analytical evaluation of the new electrochemiluminescent immunoassays (ECLIA) for TSH, FT4, and T3 in the Elecsys 2010 immunoassay system. To assess the clinical classification of patients under suspicion of thyroid disease based on these laboratory assays. MATERIALS AND METHODS: The analytical evaluation included the performance of minimum detectable concentrations, within-assay and inter-assay precision for the three analytes, functional sensitivity and linearity studies for TSH, and method comparison with the previous methods of RIA for FT4 and T3, and IRMA for TSH in current protocols of our institution. 102 patients with clinical suspicion of thyroid disease were assayed by ECLIA and radioactive techniques. Their differential clinical classification based on laboratory tests was studied as well. RESULTS: The minimum detectable concentrations coincided with the manufacturer's: <0.005 mU/L for TSH, <0.30 pmol/L for FT4, and <0.30 nmol/L for T3. Functional sensitivity for TSH was 0.044 mU/L. Over the analytical range tested, within-assay imprecision was below 3.2% for TSH, 2.2% for FT4 and 9.6% for T3, and interassay CVs were below 4.0% for TSH, 5.9% for FT4 and 12.9% for T3. Measurement of diluted sera showed the TSH assay to overestimate recoveries by 18.6%. We have compared sera results of the Elecsys ECLIA assays with those obtained from the IRMA (Spectria-Orion Diagnostica) for TSH: TSH (ECLIA) = 0.074+0.953 TSH (IRMA), (r = 0.974; Sy/x = 2.638), and RIA (Coat a Count-DPC) for FT4:FT4 (ECLIA) = 5.043+0.682 FT4 (RIA), (r = 0.770; Sy/x = 4.774) and RIA (Spectria-Orion Diagnostica) for T3: T3(ECLIA) = -0.461+1.084 T3 (RIA), (r = 0.970; Sy/x = 0.412). When sera from 102 patients were processed by both methods, minimal disagreement in the area of diagnostic classification was observed in 8/102 (7.8%) of the cases. CONCLUSION: The Elecsys 2010 is specially attractive as a routine assay because it is fully automated, obtaining results in only 18 minutes. The analytical assay performance for TSH, FT4 and T3 was shown to be acceptable. Using two different sets of diagnostic tests minimal discrepancies were found in the laboratory assessment for the classification of patients with clinical suspicion of thyroid disease. PMID- 10667474 TI - Comparison of fatty acid profiles in the serum of patients with prostate cancer and benign prostatic hyperplasia. AB - OBJECTIVE: The role of dietary fatty acids (FAs) in benign and malignant prostatic diseases was investigated by comparing the composition value of serum fatty acids in the normal controls, and patients with prostate cancer (PC) and benign prostatic hyperplasia (BPH). Also, to estimate a possible association between PC risk and PUFAs, omega-3, omega-6 and omega-3/omega-6 FA composition ratios were compared among these groups. METHODS: Serum samples were obtained from 24 BPH and 19 PC patients, and from 21 age-matched normal male subjects. The serum concentration of 21 fatty acids was determined using gas chromatography/mass spectrometry. RESULT: The proportional values of saturated fatty acids (SFAs) groups demonstrated no specific difference between the control subjects and the patients. In the polyunsaturated fatty acids (PUFAs), we found that the omega-3 PUFAs level was significantly decreased in patient with BPH and PC and that the omega-6 PUFAs level was increased in PC only. The ratio of omega 3/omega-6 PUFAs decreased in the following order of normal, BPH, and PC. CONCLUSION: It was proposed that the changed composition level of PUFAs including omega-3 and omega-6 PUFAs have certain relationship with both prostatic diseases. Therefore, the ratio of omega-3/omega-6 PUFAs also may have an important association with the benign and malignant status of prostatic disease. PMID- 10667475 TI - A splicing mutation in the hydroxymethylbilane synthase gene in a Japanese family with acute intermittent porphyria. AB - OBJECTIVES: Acute intermittent porphyria (AIP) is an autosomal dominant inherited disease caused by a decreased activity of hydroxymethylbilane synthase (HMBS). As far as the gene abnormalities of the HMBS, many different mutations have been reported. In this work, we investigated the presence of mutations in a Japanese family with AIP. DESIGN AND METHODS: A 44-year-old Japanese male and nine members of his family were investigated. All of them were screened by traditional biochemical markers. Mutational analysis was performed using polymerase chain reaction-single strand conformation polymorphism method followed by DNA sequencing. A reliable restriction enzyme cleavage assay was established for the pedigree analysis. RESULTS: The mutation was a splicing mutation, a C to G transversion at position -3 of the acceptor site of intron 11 of the HMBS gene, resulting in the exon 12 skipping. The patient is heterozygous for the mutation, and his father appeared to be the source of the mutant allele. This mutation created a new cleavage site of the Nla III restriction enzyme and could be screened by a amplified fragment from genomic DNA with digestion. Using this cleavage assay, an asymptomatic carrier in the family was definitively identified. CONCLUSIONS: This mutation was first found among Japanese AIP patients, but happened to be the same as reported previously from Europe. A similarity of gene abnormality may suggest that those European and Japanese AIP families have a common ancestor. Molecular investigations on the family members should be applied not only for more accurate diagnosis, but also for understanding the molecular genetic heterogeneity underlying this dominantly inherited enzymopathy. PMID- 10667476 TI - The effect of gestational age, birth weight, and disease on troponin I and creatine kinase MB in the first year of life. AB - OBJECTIVES: To determine the effect of gestational age and birth weight (BW) on troponin I (TnI) and creatine kinase MB fraction (CKMB) levels during the first year of life. METHODS: Troponin I and CKMB levels were determined in infants less than 1 year of age using the Immuno I (Bayer Corp.). RESULTS: Troponin I fractions were greatest in the preterm infant; the levels decreased significantly with increasing gestational age and BW, (p = 0.008 and p = 0.005, respectively). The CKMB levels did not exhibit a significant difference between the preterm and term infant groups when assessed for the effects of gestational age or BW (p = 0.12 vs. p = 0.35). Neither TnI nor CKMB levels were significantly different between preterm survivors and nonsurvivors (p = 0.31; p = 0.34, respectively). TnI levels were elevated in critically ill patients without documented myocardial infarction, and without a comparable rise in CKMB. CONCLUSION: The higher TnI levels during the first 3 months of life may indicate programmed cell death, or apoptosis. This may be especially true in the preterm infant in which the greatest values were documented. PMID- 10667477 TI - Catalase and paraoxonase in hypertensive type 2 diabetes mellitus: correlation with glycemic control. AB - OBJECTIVES: Type 2 diabetes mellitus (DM) is well recognized as being associated with increased prevalence of hypertension. Experimental and epidemiologic studies have shown that oxygen-free radicals are elevated because antioxidant enzyme activities are altered both in uncontrolled essential hypertension and DM itself. Recently paraoxonase (PON) has been recognized as an antioxidant enzyme that hydrolyzes lipid peroxides. The aim of this study is to evaluate simultaneously PON activities and antioxidant status in hypertensive type 2 DM cases and to establish any possible relationship between these parameters and duration of hypertension or diabetes, hemoglobin (Hb) A1c levels, and lipid parameters. DESIGN AND METHODS: Nineteen normotensive subjects with type 2 DM, 37 hypertensive (diastolic blood pressure 90 mm Hg or more) subjects with type 2 DM, and 25 normotensive control subjects with normal glucose tolerance were selected for this study. Superoxide dismutase (SOD), catalase, and basal-stimulated PON activities were measured by the methods of Sun et al.; Goth; and Eckerson, Wyte, and La Du, respectively; other lipid parameters were determined using an autoanalyzer. RESULTS: Catalase activities of either hypertensive patients with type 2 DM or type 2 DM patients without complication were found to be higher than controls (p<0.01), although no significant difference in SOD and basal-stimulated PON activities was observed between these groups. A significant elevation in catalase activity (p = 0.004) of patients with high HbA1c levels (>7.0%) (n = 37) compared with patients with low HbA1c levels (<7.0%) (n = 19) was detected. There was also a positive correlation between the catalase activities and fasting glucose levels and HbA1c concentrations in hypertensive patients with type 2 DM (r = 0.4567, p<0.05 and r = 0.3686, p<0.05, respectively). An increase in catalase activity of patients with B and/or AB phenotype compared with patients with A phenotype was also noted. CONCLUSION: Poor glycemic control in diabetes is strongly associated with an increase in free radicals and consequent diabetic complications. Uncontrolled glucose metabolism may also be the cause of alterations in antioxidant enzymes. Among these, catalase correlates best with poor glycemic control. The current data reveal that B allele carriers of PON are more susceptible to oxidant stress. PMID- 10667478 TI - Lipoprotein-genotype associations in Trinidadian neonates. AB - OBJECTIVES: We hypothesized that common variation in the angiotensinogen (AGT), beta-3-adrenergic receptor, intestinal fatty acid-binding protein, serum paraoxonase, paraoxonase-2, hepatic lipase, apolipoprotein E (APOE), and Werner helicase (WRN) genes would be associated with variation in biochemical phenotypes in a previously unstudied neonatal sample. DESIGN AND METHODS: We examined associations of both nongenetic and genetic variables with plasma lipoprotein traits in neonates from Trinidad. RESULTS: Among nongenetic variables, we found significant associations between plasma concentrations of: 1) lipoprotein(a) [Lp(a)] and both ethnicity (p = 0.037) and birth weight (p = 0.001); 2) total cholesterol and gender (p = 0.010); 3) triglyceride and birth weight (p = 0.035); and 4) apolipoprotein AI and gender (p = 0.016). Among genetic variables, we found that: 1) common variation on chromosome 1q in AGT codon 235 was significantly associated with variation in plasma apolipoproteins AI (p<0.0001) and B (p = 0.013); 2) common variation in WRN at codon 1367 was significantly associated with variation in plasma Lp(a) (p<0.0001); and 3) common variation in APOE at codons 112 and 158 was significantly associated with variation in plasma triglycerides (p = 0.013). CONCLUSIONS: The associations with AGT and WRN are novel and may have resulted either from a direct influence of the genetic variants or through linkage disequilibrium with other functional loci, such as the familial combined hyperlipidemia locus on chromosome 1q in the case of AGT. Despite the fact that there are some limitations in making determinations from cord blood, the results suggest that there may be genetic determinants of plasma lipoproteins in neonates. PMID- 10667479 TI - Evaluation of some tissue and serum biomarkers in prostatic carcinoma among Egyptian males. AB - OBJECTIVES: The purpose of this study is to evaluate the role of soluble E cadherin as a serum marker and bcl-2 and DNA content as tissue markers in characterization and management of prostatic adenocarcinoma (PC) among Egyptian males. DESIGN AND METHODS: The study group included 71 patients with prostatic adenocarcinoma, 30 patients with benign prostatic hyperplasia (BPH), and 20 normal male subjects. Serum soluble E-cadherin (sE-cadherin) and PSA were quantified by ELISA and MEIA (microparticle enzyme immunoassay) techniques, respectively. Tissue samples were investigated for bcl-2 chromosomal translocation t(14;18) by polymerase chain reaction (PCR) together with detection of bcl-2 protein expression by immunohistochemistry. The results were correlated with DNA content (as defined by flow cytometric analysis) and also with traditional clinicopathologic parameters. RESULTS: Our data revealed that, serum PSA was superior to sE-cadherin as a marker for PC with a sensitivity of 83% compared to 59% in case of E-cadherin at the same specificity (96.6%). Combination of both markers raised the sensitivity to 90%. E-cadherin correlated with Gleason score. Ploidy status, synthetic phase fraction (SPF), and proliferation index (PI) correlated significantly with tumor Gleason score. PI was also correlated to clinical stage. bcl-2 protein was overexpressed in 14% of PC and it showed a trend for correlation with tumor Gleason score (p = 0.06). We failed to detect chromosomal t(14;18) in the bcl-2 gene in all the studied tumors. CONCLUSIONS: E-Cadherin is a clinically useful biomarker in PC specially in combination with PSA. DNA content changes and bcl-2 oncogene may account for tumorogenesis and may assist in prognostication of PC. PMID- 10667480 TI - Effect of eating on plasma radical-trapping antioxidant activity (TRAP) in patients with cirrhosis. AB - OBJECTIVES: To ascertain the effects of eating on plasma antioxidant capacity in patients with liver disease. DESIGN AND METHODS: Eighteen cirrhotic patients were compared to 18 age and sex-matched controls. TRAP was measured by a fluorometric assay after a 12 h fast, and 60, 120, and 180 min after the study participants had taken a drink formula food. RESULTS: In the fasting state, TRAP was higher in patients with alcoholic cirrhosis (847+/-39 micromol/L, mean +/- SEM) in comparison to patients with viral cirrhosis (653+/-41) and to controls (758+/-26) (p<0.005). In cirrhotic patients, TRAP did not change in the post-absorptive state. In controls, TRAP decreased progressively, to a value of 719+/-21 (p<0.02), and the AUC of the delta-values of TRAP and of plasma insulin showed an inverse correlation (r = -0.52, p<0.05). CONCLUSIONS: In normal subjects, but not in cirrhotics, TRAP decreases in the post-absorptive state, probably in relationship with the activation of metabolic pathways. PMID- 10667481 TI - RANTES and MCP-1 chemokine plasma levels in chronic renal transplant dysfunction and chronic renal failure. AB - OBJECTIVES: Procedures to diagnose renal allograft rejection depend on detection of graft dysfunction due to the presence of mononuclear leukocytic infiltrates. DESIGN AND METHODS: In our study, we pursued an immunodiagnostic approach utilizing an ELISA method on plasma samples to monitor patients waiting to undergo transplantation in order to evidence prognostic developments in renal transplantation and, at least, to diagnose renal chronic transplant dysfunction. We analyzed blood levels of two chemokines, RANTES and MCP-1, which are normally overexpressed locally in renal chronic rejection. RESULTS: Our results showed that patients affected by chronic renal failure (and waiting for kidney transplant), as well as kidney-grafted patients affected by chronic transplant dysfunction, had plasma levels of RANTES significantly higher than those of controls (patients without acute or chronic pathologies). CONCLUSIONS: Our data suggest a simple method to evaluate the plasmatic presence of RANTES, which could be involved in longterm kidney graft failure. PMID- 10667482 TI - Effects of pentoxifylline and coenzyme Q10 in hepatic ischemia/reperfusion injury. AB - OBJECTIVES: We examined whether pentoxifylline (PTX) and coenzyme Q10 (Q) pretreatment affect ischemia-reperfusion damage in the rat liver. DESIGN AND METHODS: Twenty minutes of reflow following 30 min of ischemia was performed. Before the experiment, rats were treated PTX 50 mg/kg, IP or PTX 50 mg/kg IP + Q10 mg/kg, intragastric, or untreated. Rats were divided into four groups: control (C), ischemia-reperfusion (IR), PTX-treated (P), and Q+PTX-treated (QP) groups. Hepatic glutathione (GSH) and malondialdehyde (MDA) levels and catalase, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and reductase (GSSGR) activities were measured. RESULTS: In IR group GSH levels decreased (p<0.01), conversely MDA levels increased (p<0.01). PTX pretreatment did not affect GSH and MDA values, but Q+PTX pretreatment improved of those (p<0.01). It was shown that catalase and GSH-Px activities increased during ischemia reperfusion (p<0.01, both of), but PTX pretreatment did not significantly ameliorate those activities. GSSGR activity was higher in IR group than in basal levels (p<0.01). The decrease GSSGR activity that was observed in P group was not significant compared to IR group. During ischemia/reperfusion also SOD activity increased as compared with controls (p<0.05). In PTX-treated group, SOD activity was significantly higher than control and ischemia/reperfusion groups (p<0.01, both of). Q+PTX treatment ameliorated those enzyme activities to the control values. CONCLUSIONS: Short-term hepatic ischemia-reperfusion diminished GSH, increased MDA levels and induced some antioxidant enzyme activities. Q+PTX pretreatment was useful in hepatic ischemia-reperfusion injury, but treatment of PTX alone did not cause beneficial effect in the present study. PMID- 10667483 TI - Tumor markers in lung cancer: does the method of obtaining the cut-off point and reference population influence diagnostic yield? AB - OBJECTIVES: The aim of this study was to evaluate the clinical usefulness of the tumor markers CA125, CEA, NSE, SCC, and TPS in a group of patients with lung cancer. We estimated the influence of the method for choosing the cut-off point and of considering as a reference population either healthy controls or patients with some form of non-neoplastic pulmonary disease (NNPD). DESIGN AND METHODS: The tumor markers were determined using enzyme immunoassay techniques, and their diagnostic yield was evaluated using ROC curves and their correlation with the percentages between false and true positives. The diagnostic parameters of the tumor markers are presented in 116 patients with lung cancer and compared with a group of 25 healthy controls and another group of 80 patients with some form of NNPD. We determined on the one hand the cut-off points resulting from the best sensitivity-specificity balance in the ROC curves and on the other those resulting from considering a specificity of 95%. With the two cut-offs we studied the different diagnostic parameters: sensitivity, specificity and accuracy or area below the ROC curve. RESULTS: Optimum diagnostic yield is obtained when we choose the cut-off point determined by the best balance of sensitivity and specificity in the ROC curves and take a healthy population as a reference group. The cut-off values for CA125, CEA, NSE, SCC, and TPS were 24 U/mL, 2.8 ng/mL, 9.8 ng/mL, 1.6 ng/mL, and 67.8 U/L, respectively. CONCLUSIONS: Our results suggest that in future studies on tumor markers, a group of healthy subjects should be used as a reference population and ROC curves should be used to obtain the optimum cut-offs. PMID- 10667484 TI - Simultaneous detection of vitamin C and uric acid by capillary electrophoresis in plasma of diabetes and in aqueous humor in acute anterior uveitis. PMID- 10667485 TI - High percentage of false positive cardiac troponin I results in patients with rheumatoid factor. PMID- 10667486 TI - Fluorescent automated single-stranded conformation (F-SSCP) analysis is able to detect a point mutation at the extreme 5' end of a PCR product. PMID- 10667487 TI - Are expert systems "more intelligent" than laboratory doctors? PMID- 10667488 TI - Evaluation of the Tosoh HLC-723GHb V A1c 2.2 hemoglobin A1c analyzer. PMID- 10667489 TI - Extracellular calcineurin: identification and quantitation in serum and amniotic fluid. PMID- 10667490 TI - Serum apolipoprotein AI, B, and lipoprotein (a) levels in hypercholesterolemic schoolchildren. PMID- 10667491 TI - Modulation of neutrophil apoptosis by granulocyte colony-stimulating factor and granulocyte/macrophage colony-stimulating factor during the course of acute respiratory distress syndrome. AB - OBJECTIVE: To determine whether bronchoalveolar lavage fluid (BALF) from patients either at risk for the acute respiratory distress syndrome (ARDS) or with sustained ARDS modulates neutrophil apoptosis; to measure the BALF concentrations of the apoptosis inhibitors granulocyte colony-stimulating factor (G-CSF) and granulocyte/macrophage colony-stimulating factor (GM-CSF) before and after the onset of ARDS; and to determine whether the BALF concentrations of G-CSF and/or GM-CSF are associated with clinical outcome. DESIGN: Prospective cohort study. SETTING: Tertiary university hospital. PATIENTS: Twenty patients at risk for ARDS and 45 patients with established ARDS. INTERVENTIONS: Patients at risk for ARDS underwent bronchoalveolar lavage within 24 hrs of being identified, then again 72 hrs later. Patients with ARDS underwent bronchoalveolar lavage within 24 hrs of meeting ARDS criteria, then again on days 3, 7, and 14 of the disease. MEASUREMENTS AND MAIN RESULTS: Normal peripheral blood neutrophil were incubated overnight in BALF from normal volunteers, from patients at risk for ARDS, or from patients with ARDS. neutrophil apoptosis was determined by flow cytometric analysis of annexin V binding. G-CSF and GM-CSF were measured in BALF by immunoassays. Compared with normal BALF, BALF from patients on days 1 and 3 of ARDS inhibited neutrophil apoptosis, but BALF from patients at later stages of ARDS, or from patients at risk for ARDS, did not. The BALF concentrations of both G-CSF and GM-CSF were elevated early in ARDS and decreased toward later stages. Patients who lived had significantly higher concentrations of GM-CSF in the BALF than those who died. CONCLUSIONS: We conclude that the antiapoptotic effect of ARDS BALF on normal neutrophil is highest during early ARDS, and decreases during late ARDS. G-CSF and GM-CSF are present in BALF from patients with ARDS, and their concentrations parallel the antiapoptotic effect of ARDS BALF. These data support the concept that the life-span of neutrophil in the air spaces is modulated during acute inflammation. GM-CSF in the air spaces is associated with improved survival in patients with ARDS. PMID- 10667492 TI - Prevention of infection in multiple trauma patients by high-dose intravenous immunoglobulins. AB - OBJECTIVE: To investigate the activity of intravenous immunoglobulin (IVIG) as a prophylactic agent against infection in trauma victims. DESIGN: Prospective, randomized, double-blind, placebo-controlled study. SETTING: A 20-bed university intensive care unit. PATIENTS: Thirty-nine trauma patients with injury severity scores (ISSs) of 16-50. INTERVENTIONS: Penicillin was given at the time of admission and continued at least until day 4. Twenty-one patients received IVIG and 18 patients received human albumin at 1 g/kg in four divided doses (days 1, 2, 3, and 6). The two groups had similarities in age, gender, Acute Physiology and Chronic Health Evaluation II score, risk of death, and Glasgow Coma Scale score, but differing ISSs (p = .02), at the time of admission. Blood was collected on days 1, 4, and 7. MEASUREMENTS AND MAIN RESULTS: Clinical variables related to infection were recorded. The complement components C3c, C4 and CH50, IgG, and the fractions of IgG were measured. The serum bactericidal activity (SBA) was assessed at 37 degrees C (98.6 degrees F) and 40 degrees C (104.0 degrees F) at the time of admission and during the course of IVIG administration. Controlling for ISS, IVIG-treated patients had fewer pneumonias (p = .003) and total non-catheter-related infections (p = .04). Catheter-related infections (p = .76), length of stay in the intensive care unit, antibiotic days, and infection related mortality did not differ between the two groups. A significantly increased trend in IgG and its subclasses was shown on days 4 and 7 in the IVIG group but not in the control group (p<.000001). No important differences were noted in complement fractions. The SBA of the groups was similar on day 1, but significantly higher on days 4 and 7 (p<.000001) in the IVIG group, remaining so controlling for complement and ISS. SBA was higher at 40 degrees C (104.0 degrees F) compared with 37 degrees C (98.6 degrees F) (p<.0001) under all three conditions. In both groups, low SBA (on days 1, 4, and 7) was associated with increased risk of pneumonia (p<.01) and non-catheter-related infections (p = .06 for day 1; p<.01 for days 4 and 7). CONCLUSIONS: Trauma patients receiving high doses of IVIG exhibit a reduction of septic complications and an improvement of SBA. Early SBA measurement may represent an index of susceptibility to infection. PMID- 10667493 TI - Changes in the management of severe traumatic brain injury: 1991-1997. AB - OBJECTIVE: To survey the management of head-injured patients in 1997 and to identify differences compared with a survey conducted in 1991. DESIGN: A two-page questionnaire was mailed to all neurosurgeons in North America certified by the American Board of Neurologic Surgeons, asking their views regarding the most appropriate acute care of patients with severe traumatic brain injury (TBI). SETTING: North American neurosurgical practices. PATIENTS: Not applicable. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Compared with a 1991 survey, there was a significant increase in the proportion of neurosurgeons who felt these patients should have intracranial pressure monitoring (28% vs. 83%) and a decrease in the proportion who used prophylactic hyperventilation therapy (83% vs. 36%) and steroids (64% vs. 19%). Ninety-seven percent of respondents felt that the cerebral perfusion pressure should be maintained at >70 mm Hg, and 44% indicated that patients with severe TBI should be treated at Level I trauma centers. CONCLUSIONS: There have been significant changes in the acute management of patients with severe TBI since 1991. Current practices more closely reflect the recommendations of evidence-based guidelines. PMID- 10667494 TI - Survivors of catastrophic illness: outcome after direct transfer from intensive care to extended care facilities. AB - OBJECTIVE: To describe outcomes of adult survivors of prolonged critical illness after direct transfer to extended care facilities. DESIGN: A retrospective cohort study. SETTING: All adult intensive care units (ICUs) in a tertiary care university hospital. PATIENTS: A consecutive series of 97 adult survivors with an ICU stay of > or =7 days transferred directly from intensive care to extended care facilities between 1990 and 1996. INTERVENTIONS: None. METHODS AND MAIN RESULTS: Hospital and extended care facility charts were reviewed for patient characteristics, resource utilization, and survival. Survivors were for a minimum of 1 yr and a maximum of 6 yrs, and were interviewed to assess quality of life and functionality. The mean age of the patients was 66+/-16 (range, 19-93) yrs. The median length of ICU stay for these patients was 39 (range, 7-276) days. Only 18 of the 71 ventilator-assisted patients were weaned from mechanical ventilation after transfer to the extended care facility. Survival for the study period, at 1 yr after discharge from the ICU, was 49.5%. One year after discharge from the ICU, 11.5% of all patients had returned home, were breathing spontaneously, had a fair or better quality of life, and had good physical functionality. Each successive year, an increasing proportion of patients underwent direct transfer to an extended care facility. This strategy decreased the patients' length of stay (p<.002) in the ICU from year to year, but was significantly associated with an increase in readmissions to acute care hospitals (p<.002). CONCLUSIONS: Survivors of catastrophic illness who are so debilitated that they require transfer to an extended care facility have a low likelihood of achieving both survival and functional independence 1 yr after discharge from the ICU. Aggressive cost-conscious strategies to accelerate the transfer of these patients successfully reduced the length of ICU stay and hospital costs, but were associated with a high rate of readmission to tertiary care facilities. PMID- 10667495 TI - Comparison of acute physiology and chronic health evaluations II and III and simplified acute physiology score II: a prospective cohort study evaluating these methods to predict outcome in a German interdisciplinary intensive care unit. AB - OBJECTIVE: To evaluate the ability of three scoring systems to predict hospital mortality in adult patients of an interdisciplinary intensive care unit in Germany. DESIGN: A prospective cohort study. SETTING: A mixed medical and surgical intensive care unit at a teaching hospital in Germany. PATIENTS: From a total of 3,108 patients, 2,795 patients (89.9%) for Acute Physiology and Chronic Health Evaluation (APACHE) II and 2,661 patients (85.6%) for APACHE III and Simplified Acute Physiology Score (SAPS) II could be enrolled to the study because of defined exclusion criteria. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Probabilities of hospital death for patients were estimated by applying APACHE II and III and SAPS II and compared with observed outcomes. The overall goodness-of-fit of the three models was assessed. Hospital death rates were equivalent to those predicted by APACHE II but higher than those predicted by APACHE III and SAPS II. Calibration was good for APACHE II. For the other systems, it was insufficient, but better for SAPS II than for APACHE III. The overall correct classification rate, applying a decision criterion of 50%, was 84% for APACHE II and 85% for APACHE III and SAPS II. The areas under the receiver operating characteristic curve were 0.832 for APACHE II and 0.846 for APACHE III and SAPS II. Risk estimates for surgical and medical admissions differed between the three systems. For all systems, risk predictions for diagnostic categories did not fit uniformly across the spectrum of disease categories. CONCLUSIONS: Our data more closely resemble those of the APACHE II database, demonstrating a higher degree of overall goodness-of-fit of APACHE II than APACHE III and SAPS II. Although discrimination was slightly better for the two new systems, calibration was good with a close fit for APACHE II only. Hospital mortality was higher than predicted for both new models but was underestimated to a greater degree by APACHE III. Both score systems demonstrated a considerable variation across the spectrum of diagnostic categories, which also differed between the two models. PMID- 10667496 TI - Acute quadriplegia and loss of muscle myosin in patients treated with nondepolarizing neuromuscular blocking agents and corticosteroids: mechanisms at the cellular and molecular levels. AB - OBJECTIVE: Long-term treatment with nondepolarizing neuromuscular blocking agents and corticosteroids in the intensive care unit is not benign, and an increasing number of patients with acute quadriplegic myopathy have been reported with increased use of these drugs. The purpose of this study was to investigate the mechanisms underlying acute quadriplegic myopathy. DESIGN: Percutaneous muscle biopsy samples were obtained, and electrophysiologic examinations were performed during the acute phase and during recovery in patients with acute quadriplegic myopathy. Regulation of muscle contraction and myofibrillar protein synthesis was studied using cell physiologic techniques, ultrasensitive electrophoresis, in situ hybridization, and histopathologic techniques. SETTING: All patients were seen in the intensive care unit of different university hospitals. PATIENTS: All patients were critically ill with sepsis. They had been given massive doses of corticosteroids in combination with variable doses of neuromuscular blocking agents. All patients developed paralysis of spinal nerve-innervated muscles. On the other hand, cranial nerve-innervated muscle and sensory and cognitive functions were well maintained after discontinuation of treatment with neuromuscular blocking agents. INTERVENTION: Muscle biopsy samples were obtained and electrophysiologic examinations were performed in all patients. MEASUREMENTS AND MAIN RESULTS: The major observations in patients with acute quadriplegic myopathy were, as follows: a) a general decrease in myofibrillar protein content; b) specific but highly variable partial or complete loss of myosin and myosin associated proteins; c) very low thick-filament/thin-filament protein ratios; d) absence of myosin messenger RNA; and e) a dramatically impaired muscle cell force generating capacity in the acute phase of acute quadriplegic myopathy. During clinical improvement, normal expression of myosin messenger RNAs, reexpression of thick-filament proteins, and increased specific tension were observed. CONCLUSIONS: Acute quadriplegic myopathy is associated with a specific decrease in thick-filament proteins related to an altered transcription rate. Although the decreased content of thick-filament proteins is important for prolonged muscle weakness, it is not the primary cause of muscle paralysis in the acute stage, during which impaired muscle membrane excitability probably plays a more significant role. Several factors contribute to this condition, but the action of corticosteroids seems to be the predominant one, along with potentiation by neuromuscular blocking agents, immobilization, and probably also concurrent sepsis. PMID- 10667497 TI - Characterization of gastrointestinal bleeding in severely ill hospitalized patients. AB - OBJECTIVE: To characterize the source of bleeding and the prognosis in critically ill patients with upper gastrointestinal hemorrhage that developed while in the hospital. SETTING: Intensive care units of a large academic tertiary-care center. DESIGN: Retrospective cohort study. SUBJECTS: Patients undergoing endoscopy in intensive care units for gastrointestinal bleeding that developed while in the hospital. MEASUREMENTS AND MAIN RESULTS: Medical records were available for 142 patients. Of these, 66 met the criteria for in-hospital bleeding. Peptic ulcer disease, present in 56% of patients, was the most common bleeding source identified. Of patients with peptic ulcer disease, nine of 37 (24%) had stigmata of recent hemorrhage. Ten patients (15%) received endoscopic hemostasis interventions (eight receiving therapy for bleeding ulcers, two receiving therapy for esophageal varices). The in-hospital mortality rate was 42%. The cause of death was sepsis and/or multiple system organ failure in 21 patients (75%); the gastrointestinal bleeding may have contributed to the onset of sepsis in one of these patients. No patients died directly of gastrointestinal bleeding. CONCLUSIONS: Critically ill patients who bleed while in the hospital have similar sources of bleeding and rates of endoscopically directed therapy as patients admitted to hospital with bleeding. The mortality rate is very high in patients with bleeding that develops in the hospital, and this is usually a result of systemic disease. These data may help clinicians and patients to estimate the potential benefit of urgent endoscopy in critically ill patients. PMID- 10667498 TI - Thermodilution versus inert gas rebreathing for estimation of effective pulmonary blood flow. AB - OBJECTIVE: To compare measurements of the effective pulmonary blood flow (Qep, i.e., nonshunted fraction of cardiac output, Qt) by the inert gas rebreathing (RB) method and the thermodilution (TD) technique in critically ill patients. DESIGN: Prospective, comparative study of a noninvasive method and an established invasive technique. SETTING: An 11-bed general intensive care unit in a university hospital. PATIENTS: A total of 14 critically ill patients, all mechanically ventilated and monitored with systemic and pulmonary artery catheters. MEASUREMENTS AND MAIN RESULTS: Qep was determined in duplicate by RB using a mass spectrometer for gas analysis. For each determination, Qt was measured in triplicate by the cold water bolus TD technique and averaged. Simultaneously mixed venous and arterial blood samples were analyzed to calculate the intrapulmonary shunt fraction and thereby convert estimates of Qt to Qep. Mean difference between paired estimates (RB - TD) was 0.01 L/min, so for differences was 1.19 L/min, and 95% confidence interval for the bias was -0.45 to 0.47 L/min. Coefficients of variation for repeated Qep estimates were 8% (RB) and 12% (TD), respectively. Coefficients of variation for RB estimates of functional residual capacity and lung tissue volume were 6% and 17%, respectively. CONCLUSIONS: The RB method is a promising method for simultaneous noninvasive estimation of Qep and functional residual capacity in mechanically ventilated patients. However, further investigations are needed to evaluate potential problems of the method before it can be recommended for clinical purposes. PMID- 10667499 TI - Evolution of leukotriene B4, peptide leukotrienes, and interleukin-8 plasma concentrations in patients at risk of acute respiratory distress syndrome and with acute respiratory distress syndrome: mortality prognostic study. AB - OBJECTIVE: To compare the evolution of plasma concentrations of leukotriene (LT) B4, LTC4, LTD4, and interleukin (IL)-8 in patients with acute respiratory distress syndrome (ARDS) and in patients at risk of ARDS and to assess the value of these mediators in predicting mortality rate from ARDS. DESIGN: A case-control study comparing ARDS patients and patients at risk of ARDS as well as survivors and nonsurvivors with ARDS. SETTING: Hospital intensive care unit, laboratory, and department of hematology. PATIENTS: Twenty-one patients with ARDS and 14 patients at risk of ARDS. INTERVENTION: Arterial blood samples were collected on days 0, 1, and 5 after admission to the intensive care unit. MEASUREMENTS AND MAIN RESULTS: LTs were extracted, separated by high-pressure liquid chromatography and quantified by enzyme immunoassay. IL-8 was analyzed by ELISA. Plasma concentrations of LTB4 and LTC4 plus LTD4 were significantly higher in ARDS patients than in patients at risk of ARDS during the first 24 hrs. Concentrations of IL-8 were also higher in ARDS patients than in patients at risk throughout the study, although the differences between the two groups were only significant on day 5. Only the plasma concentration of LTB4 on day 1 was a marker of ARDS (72.2% sensitivity, 84.6% specificity). A logistic regression analysis showed that LTB4 and IL-8, on day 1, were markers of mortality rate in patients with ARDS (70.0% sensitivity, 87.5% specificity). CONCLUSIONS: LTs are elevated during the early phases of ARDS, whereas IL-8 increases throughout the study. The evaluation of LTB4 and IL-8 may be useful prognostic indices in patients with early phase ARDS after admission to the intensive care unit. PMID- 10667500 TI - Accidental removal of endotracheal and nasogastric tubes and intravascular catheters. AB - OBJECTIVES: To characterize the rates of accidental removal of endotracheal tubes, nasogastric tubes, central venous catheters, and arterial catheters. To assess the efficacy of corrective measures aimed at reducing the accidental removal of these devices. DESIGN: Prospective, observational, and interventional study. SETTING: Eighteen-bed medical-surgical intensive care unit of a 650-bed tertiary care hospital. PATIENTS: Patients admitted to the intensive care unit who had any of the following devices in place for more than 24 hrs: endotracheal tube, nasogastric tube, central venous catheter, arterial catheter. MEASUREMENTS AND INTERVENTIONS: Data were collected on the date of placement of tubes and catheters, position of vascular catheters, date of removal, and reason for removal. The study involved three consecutive 6-month periods. At the end of the first and the second periods, information about rates of accidental removal was provided to the physicians and nurses. In addition, the personnel were instructed to be more vigilant and specific measures aimed at reducing the accidental removal were introduced. MAIN RESULTS: In the first period, 289 endotracheal tubes were placed and 13.1% (24.7 per 1000 days) were removed accidentally. In the second and third periods, 17.1% (25.5 per 1000 days) and 11.4% (15.1 per 1000 days) were removed accidentally, respectively. In the first period, 368 nasogastric tubes were placed and 41% (73.9 per 1000 days) were removed accidentally. In both the second and the third period, a significant reduction in the rate of accidental removal was observed (32.4% or 41.2 per 1000 days and 25.8% or 29.8 per 1000 days, respectively). A significant decrease was observed in the rates of accidental removal of central venous catheters from 7.5% (12.4 per 1000 days) in the first period to 3.6% (5.4 per 1000 days) in the second period. The rate of arterial catheters accidentally removed expressed according to the time at risk significantly decreased from 46.5 per 1000 days in the first period to 19.1 per 1000 days in the second period and 25.3 per 1000 days in the third period. CONCLUSIONS: The information provided by the rates of accidental removal expressed by patient-days is helpful to compare results obtained in populations with different times of follow-up. Education of medical personnel and limiting upper-extremity access to within 20 cm from any catheter or tube resulted in a significant reduction of patient-related removal of tubes and catheters. PMID- 10667501 TI - Bacterial contamination of ready-to-use 1-L feeding bottles and administration sets in severely compromised intensive care patients. AB - OBJECTIVE: In intensive care patients, enteral feeding requires sterile feedings because of infectious complications and adequate supplements to meet nutritional needs. Heretofore, prepacked, large-volume formula containers were developed, but bacterial contamination occurred in 4% to 15%. Our objective was to investigate the microbial contamination rate of 1-L feeding bottles and newly designed administration sets over hanging times of 24 hrs in the intensive care unit (ICU). DESIGN AND SETTING: A prospective observational cohort study of patients admitted to the ICU of a university hospital. PATIENTS: All consecutive patients fed via a nasojejunal tube for at least 4 days. MEASUREMENTS: Cultures of feeding bottles, administration sets, and gastric and tracheobronchial aspirates at day 0, 1, 2, 4, and 7. RESULTS: A total of 4% of feeding bottles and 74% of infusion sets contained >10(2) colony forming units (CFU)/mL. Gastric and bronchial aspirates were positive in 90% and 92%, respectively. Bacterial counts of feeding bottles were 10(2)-10(5) CFU/mL, and the main bacteria isolated included Enterobacter cloacae, Klebsiella oxytoca, and enterococci. One third of all cultured bacteria in feeding bottles, administration sets, stomach, and lungs belonged to the Enterobacteriaceae family, which was held responsible for the nosocomial infections in the ICU. None of the 1-L feeding bottles with a hanging time of 19-24 hrs was contaminated. Only bottles that had to be exchanged because of need for a faster rate of infusion proved to be contaminated, apparently without clinical consequences. With time and the increasing severity of disease, the administration sets became contaminated at an increasingly faster rate and with higher bacterial counts mainly through retrograde growth of endogenous bacteria. The final step of bottle contamination might have been the bacterial transfer by nurses' hands. CONCLUSION: Despite an almost ideal design of the enteral nutrition delivery system, a 4% contamination rate of initially sterile feedings with clinically relevant bacteria and the fact that only manipulated systems showed bacterial growth are of concern. PMID- 10667502 TI - Human neutrophil activation and increased adhesion by various resuscitation fluids. AB - OBJECTIVE: To determine whether activated neutrophils play a major role in secondary tissue injury after resuscitation in trauma. We hypothesized that human neutrophil activation and adhesion vary, depending on the type and amount of resuscitation fluid used. SETTING: University-based research facility. SUBJECTS: Ten healthy adult volunteers. DESIGN: Whole blood from volunteers was serially diluted in polypropylene tubes with various resuscitation fluids. Fluids tested were phosphate-buffered saline, normal saline, lactated Ringer's solution, dextran, hespan, 5% human albumin, 25% human albumin, 3.5% hypertonic saline, and 7.5% hypertonic saline. Neutrophil activation (intracellular oxidative burst activity with dichlorofluorescin diacetate staining) and adhesion (integrin cell surface expression of CD18) were measured with flow cytometry (fluorescence activated cell sorting). Blood was diluted with hypertonic saline by controlling for sodium content equal to normal saline. dose-related increase in neutrophil oxidative burst activity as the result of dilution followed with crystalloid fluids and artificial colloids (dextran and hespan). The increase was 12-18 x baseline at the 75% dilution. The increase with 5% human albumin was only 2.2 x baseline, and 25% albumin did not demonstrate any increased intracellular activity. A similar significant increase in the neutrophil adhesion expression (CD18) occurred with artificial colloids (p<.05) and, to a lesser extent, with crystalloids, but not with albumin. Hypertonic saline caused a decrease in CD18 cell surface expression. CONCLUSIONS: This study suggests that the neutrophil activation and adhesion may vary, depending on the type of resuscitative fluid used. All artificial resuscitative fluids may not be similar or innocuous, as demonstrated by the dose-related increase in neutrophil activation and adhesion. PMID- 10667503 TI - Interhospital transport of the extremely ill patient: the mobile intensive care unit. AB - BACKGROUND: Critically ill patients may require specialized care that is offered only at tertiary referral centers. As regionalization and specialization of critical care become more common, transportation of critically ill patients must be refined. Transportation of critically ill patients within a hospital, much less outside the hospital, is often deemed unsafe because of medical instability. We report, here, our results from 2 yrs' experience of transporting extremely ill patients with respiratory failure via a ground critical care transport service. METHODS: A mobile intensive care unit was equipped and staffed to nearly recreate the intensive care environment. Staffing included a physician, nurse, respiratory therapist, and driver--all with extensive critical care experience. The mobile intensive care unit was equipped with a full pharmacy, advanced ventilatory equipment, and capability for full invasive hemodynamic monitoring. Data were analyzed by retrospective review. The predicted mortality rate, based on Pao2/Fio2 ratios, was compared with the actual mortality rate. RESULTS: During a 2-yr period, 39 critically ill patients were transported. Thirty-six of the 39 were candidates for extracorporeal lung assist, with a mean positive end expiratory pressure requirement of 15.9, a mean Fio2 requirement of .93, and a mean Pao2/Fio2 ratio of 59.8. Pulmonary arterial catheters and peripheral arterial catheters were in place in 66.6% and 72% of patients, respectively. Vasoactive medications were being infused in 56%, and 74% were receiving medical paralytics. One patient died during movement from the bed to the transport gurney. Other than one episode of transient hypotension, there were no complications or untoward outcomes related to transport. Unique therapeutic interventions were performed at the receiving facility on 34 of 39 patients. The predicted mortality rate, based on indicators of lung dysfunction, was 68% to 100%; the actual subsequent hospital mortality rate was 43%. CONCLUSIONS: When a mobile intensive care unit is properly staffed and equipped and patient stabilization is performed before transfer, severely ill patients with respiratory failure can be transferred safely. For patients with respiratory failure, there may be a survival advantage in transfer to regional centers of expertise. PMID- 10667504 TI - Early psychological reactions to life-threatening injuries. AB - OBJECTIVE: To assess the prevalence of posttraumatic stress symptoms and coping patterns in severely injured accident victims; to study correlations between injury severity and psychosocial variables and the presence of posttraumatic stress symptoms; and to analyze intensive care unit (ICU) personnel's global clinical appraisals in relation to patient characteristics. DESIGN: A study of critically ill accident victims assessed within one month of the trauma. SETTING: ICU of the traumatology department at the University Hospital, Zurich. PATIENTS: 121 consecutive patients with accidental injuries (mean Injury Severity Score, 21.8; mean Glasgow Coma Scale score, 14.4) admitted to the ICU between January 1996 and June 1997, aged 18-68 yrs. Patients with severe head injuries, attempted suicides, and victims of physical assault were excluded. MEASUREMENTS: Extensive clinical interview, Impact of Event Scale, Clinician-Administered Posttraumatic Stress Disorder Scale, social support, life events, biographical protective and risk factors, Sense of Coherence questionnaire, Freiburg Questionnaire of Coping with Illness. RESULTS: 13.7 (SD, 6.8) days after the accident, 5 patients (4.1 %) met all criteria for posttraumatic stress disorder with the exception of the time criterion. A further 24 patients (19.9%) had subsyndromal posttraumatic stress disorder. Posttraumatic psychiatric symptomatology did not correlate with objective injury criteria, but rather with pretrauma variables (female gender, biographical risk and protective factors, life events), the patients' subjective appraisal of the severity and threat of the accident, their general attitude toward life ("sense of coherence"), and their current coping strategies. Surgeons' and nurses' global clinical appraisals did not correlate with injury severity or with the patients' coping strategies. CONCLUSIONS: Trauma surgeons and ICU personnel should pay special attention to the strains and stressors their patients have been exposed to when recording case histories and to the level of their patients' psychosocial adaptation before the trauma. PMID- 10667505 TI - Hypocalcemia and parathyroid hormone secretion in critically ill patients. AB - OBJECTIVE: To investigate possible causes of hypocalcemia and to assess parathyroid hormone (PTH) secretion in intensive care unit (ICU) patients. DESIGN: Combined cross-sectional and prospective study. SETTING: ICU in a university hospital. PATIENTS: Thirteen patients with sepsis and 13 patients who underwent major surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Calcium metabolic indices were investigated during the first 24 hrs in the ICU and after 2 days. Eight of the surgical patients and five of the septic patients were subjected to a citrate/calcium infusion on day 1 in the ICU, to study the dynamics of PTH secretion. The blood ionized calcium (Ca2+) concentration was generally low in the septic patients (mean +/- SD, 1.03+/-0.08 mmol/L; reference value, 1.10-1.30) and increased, but not normalized, after 2 days. Hypocalcemia was only occasionally seen in the surgical patients. In the septic patients, urinary excretion of calcium was low; and, in both patient groups, elevated concentrations of two markers of bone resorption, deoxypyridinoline and ICTP (serum carboxy-terminal cross-linked telopeptide of type I collagen), were found. In cases of sepsis, the concentrations of proinflammatory cytokines were high (394+/-536 pg/mL for tumor necrosis factor-alpha and 5676+/-5190 pg/mL for interleukin-6, both normally <10-20). The Ca2+ concentration was inversely related to tumor necrosis factor-alpha and interleukin-6 (r2 = .35-.42; p<.01), as well as to procalcitonin (r2 = .71; p<.01). Despite normocalcemia in the surgical patients, serum PTH concentrations were elevated in both patient groups (97 and 109 ng/L) (reference value, <55 ng/L), both on day 1 and day 3 in the ICU. The citrate/calcium infusion revealed an increased secretory response of PTH to lowered Ca2+ concentrations in both groups of patients (p<.05), when compared with matched healthy controls. CONCLUSION: Hypocalcemia was common in septic ICU patients and was not the result of an increased urinary excretion of calcium or of an attenuated bone resorption, but seemed related to the inflammatory response. An increased PTH secretion was found in both patient groups. PMID- 10667506 TI - Death in two Canadian intensive care units: institutional difference and changes over time. AB - OBJECTIVE: To study and compare the mode of death in two different institutions' intensive care units (ICUs) for the two time periods, 1988 and 1993. DESIGN: Retrospective chart review. SETTING: Medical/surgical/trauma ICUs in two tertiary care teaching hospitals. PATIENTS: Patients dying in the medical/surgical/trauma ICUs between January 1, 1988 and December 31, 1988; and January 1, 1993 and December 31, 1993. Data collection included demographics, origin of admission, date of ICU admission, date of death, Acute Physiology and Chronic Health Evaluation (APACHE) III diagnostic categories, APACHE II physiologic variables, organ system failures present at the time of admission and 24 hrs before death, and mode of dying. APACHE II scores and mortality risk were calculated. Data analysis included a multiple analysis of variance to assess overall effect, with subsequent analyses of variance to assess the effect of institution and year on each individual dependent variable. All results are reported as mean +/- SEM values. RESULTS: A total of 439 charts were reviewed. Gender, age, and origin of admission were not different between the 2 yrs or the two institutions. Mean APACHE II scores and organ system failures were lower at Hospital A in 1998 vs. Hospital B, as was predicted mortality. These factors increased at Hospital A in 1993 and were similar to those at Hospital B. Withdrawal of support was much more common in 1993 than 1988 at both institutions (43% at Hospital A and 46% at Hospital B in 1988 vs. 66% at A and 80% at B in 1993), increasing to a greater extent in 1993 at Hospital B (p<.05). Length of stay in the ICU was significantly longer at Hospital A than at Hospital B in 1988 (9.4+/-1.4 vs. 4.3+/-0.6 days; p<.05) and in 1993 (8.2+/-2.9 vs. 3.8+/-0.5 days; p < .05). CONCLUSIONS: There has been an increase in withdrawal of life support, in recent years, at both the institutions studied. Differences exist between institutions with respect to end of-life decisions in the ICU. These differences are likely representative of widely prevalent regional differences and are the result of many factors. PMID- 10667507 TI - Lipopolysaccharide binding protein in acute pancreatitis. AB - OBJECTIVE: To assess the expression of plasma lipopolysaccharide binding protein (LBP) concentrations and its relationship to markers of the systemic inflammatory response syndrome during acute pancreatitis. DESIGN: A prospective study. SETTING: General surgical units of university teaching hospitals in the Belfast area. PATIENTS: The study included 18 patients admitted with established diagnosis of acute pancreatitis on the basis of elevated serum amylase or by contrast radiology. Patients were retrospectively stratified using the Modified Glasgow Criteria into severe (n = 7) and mild (n = 11) disease. INTERVENTIONS AND MEASUREMENTS: Blood samples were obtained at admission (day 1) and for a further 3 days for the measurement of LBP, C-reactive protein (CRP), tumor necrosis factor, and interleukin (IL)-6. Acute Physiology and Chronic Health Evaluation (APACHE) II scores were calculated on day 1 and day 2. MAIN RESULTS: LBP and CRP concentrations were significantly increased from healthy control values in acute pancreatitis patients at presentation. In the mild group LBP, CRP and IL-6 concentrations remained relatively constant throughout the study period. By comparison, severe acute pancreatitis was associated with significantly higher LBP concentrations and a marked systemic inflammatory response as evidenced by increased CRP, IL-6, and APACHE II scores. The rise in LBP occurred after the observed increase of these markers. Significant correlations were found among CRP and LBP, IL-6 and LBP, and IL-6 and APACHE II scores. There were no fatalities in the mild group, whereas four of the seven patients with severe disease died. CONCLUSIONS: LBP was significantly raised in patients with severe acute pancreatitis but would seem to be of limited use in predicting disease severity. This acute phase protein may have a role in the progression of systemic complications associated with acute pancreatitis. PMID- 10667508 TI - Helicobacter pylori infection: a risk factor for upper gastrointestinal bleeding after cardiac surgery? AB - OBJECTIVE: To determine the prevalence of Helicobacter pylori (H. pylori) in critically ill patients who develop upper gastrointestinal bleeding after cardiac surgery in relation to other risk factors. DESIGN: Prospective, single center, cohort study. SETTING: Surgical intensive care unit in a university hospital. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Over a 1-yr period, all consecutive patients with upper gastrointestinal hemorrhage from the stomach or duodenum were studied for H. pylori infection by serology. Additionally, the need for mechanical ventilation over 48 hrs, the duration cardiopulmonary bypass, and the aortic cross-clamp time were analyzed. For control, 229 patients with no evidence of gastrointestinal hemorrhage after cardiac surgery were studied. All patients received stress ulcer prophylaxis with ranitidine. Operations were performed on 2,956 patients during the study period. The incidence of upper gastrointestinal bleeding was 0.9%. Twenty (77%) of the 26 patients with upper gastrointestinal bleeding and 145 (63%) patients of the control group had serologic evidence for H. pylori infection (odds ratio, 1.9; 95% confidence interval 0.7-5.0; p = .2). Patients who required prolonged mechanical ventilation had a significantly greater risk for upper gastrointestinal bleeding (odds ratio, 22.1; 95% confidence interval 8.6-56.7; p<.001). Patients with upper gastrointestinal bleeding also had a significantly longer duration of cardiopulmonary bypass and aortic cross-clamp time (p<.001) CONCLUSIONS: H. pylori is not associated with upper gastrointestinal bleeding in critically ill patients who receive stress ulcer prophylaxis, whereas patients who require prolonged mechanical ventilation are at high risk. A prophylactic eradication of H. pylori is not justified. PMID- 10667509 TI - Evolution of lactate/pyruvate and arterial ketone body ratios in the early course of catecholamine-treated septic shock. AB - OBJECTIVES: To measure arterial lactate/pyruvate (L/P) and arterial ketone body ratios as reflection of cytoplasmic and mitochondrial redox state at different stages of catecholamine-treated septic shock and compare them with normal and pathologic values obtained in patients in shock who have decreased oxygen transport (cardiogenic shock), and to assess the relationship between the time course of lactate, L/P ratio, and mortality in septic shock. DESIGN: Prospective, observational human study. SETTING: A university intensive care unit. PATIENTS: Sixty consecutive adult patients who developed septic shock and lactic acidosis requiring the administration of vasopressors. Twenty patients in the intensive care unit without shock, sepsis, and hypoxia and with normal lactate values and 10 patients with cardiogenic shock were also studied. MEASUREMENTS: Hemodynamic measurements, arterial and mixed venous blood gases, arterial lactate and pyruvate concentrations, and arterial ketone body ratio were measured within 4 hrs after the introduction of catecholamine and 24 hrs later. MAIN RESULTS: Fifteen patients (25%) died within the first 24 hrs of septic shock, and these early fatalities had a higher blood lactate (12.2+/-3 versus 4.6+/-1.3 mmol/L; p<.01) concentration and a higher L/P ratio (37+/-4 versus 20+/-1; p<.01) than those who died later. No difference was found for arterial ketone body ratio (0.41+/-0.1 versus 0.50+/-0.06). Forty-five patients survived >24 hrs including 25 survivors and 20 nonsurvivors. Although there was no difference between survivors and nonsurvivors in initial lactate concentration (4.1+/-0.4 and 4.6+/ 0.3, respectively), L/P ratio (19+/-1 and 20+/-1, respectively), and arterial ketone body ratio (0.5+/-0.06 and 0.52+/-0.07, respectively), blood lactate and L/P ratio significantly decreased during the first 24 hrs in the survivors (2.8+/ 0.4 and 14+/-1, respectively; p<.05). and were stable in the nonsurvivors (4+/ 0.3 and 22+/-1, respectively) Although returning to normal values after 24 hrs in survivors and nonsurvivors, arterial ketone body ratio was higher in survivors (1.72+/-0.17 versus 1.09+/-0.15; p<.05). Lactate and L/P ratio were closely correlated (r2 = .8, p<.0001). In the cardiogenic shock group, lactate concentration was 4+/-1 mmol/L, L/P ratio was 40+/-6, and arterial ketone body ratio was 0.2+/-0.05. The mortality rate was 60%. CONCLUSIONS: The main result of the present study is that hemodynamically unstable patients with sepsis needing catecholamine therapy had a lactic acidosis with an elevated L/P ratio and a decreased arterial ketone body ratio, suggesting a decrease in cytoplasmic and mitochondrial redox state. The duration of lactic acidosis is associated with the development of multiple organ failure and death. PMID- 10667510 TI - Intensive care unit morbidity and mortality from eclampsia: an evaluation of the Acute Physiology and Chronic Health Evaluation II score and the Glasgow Coma Scale score. AB - OBJECTIVE: To determine the maternal morbidity and mortality in patients with eclampsia admitted to an intensive care unit (ICU), and to establish the efficacy of the Acute Physiology and Chronic Health Evaluation (APACHE) II score, the organ system failure score as defined by Knaus, and the Glasgow Coma Scale (GCS) score in predicting outcome. DESIGN: Retrospective analysis of a 3.5-yr period. SETTING: Surgical ICU in a university hospital. PATIENTS: A total of 105 patients who were admitted with a diagnosis of eclampsia were studied. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The data captured included the reason for admission, maternal age, gestational age, parity, number of seizures, duration of ICU stay, anticonvulsant therapy, drug therapy, GCS score, APACHE II score, and the occurrence of organ failure. Of the 126 patients with eclampsia who were admitted to the ICU, records of 105 patients (83%) were found. The overall mortality was 10.5% (n = 11). The mean age, gestation, parity, number of preadmission seizures, and duration of stay were similar in survivors and nonsurvivors. Although the APACHE II score was significantly higher in nonsurvivors, multiple logistic regression analysis suggested that the goodness of-fit scores for GCS and APACHE II were similar (38.29 vs. 38.01). The GCS scores of survivors were significantly higher than those of nonsurvivors (10.61 vs. 5.0; p<.001). Respiratory failure was the most common organ failure in both groups. The mean number of organ failures was higher in nonsurvivors compared with survivors (2.9 vs. 1.3; p<.001). An occurrence of more than two organ failures that persisted for >48 hrs was invariably associated with a fatal outcome. Anticonvulsant therapy consisted of magnesium sulfate or phenytoin and a midazolam infusion. Only one patient (0.9%) had a seizure, and this occurred en route to the ICU. No seizures occurred after admission to the ICU. CONCLUSIONS: The organ system failure score and the GCS score are good predictors of outcome in eclampsia. Apart from the GCS score, other variables in the APACHE II score are not valuable for outcome prediction. The low GCS score in nonsurvivors suggests that closer attention to the neurologic management may be beneficial. A prospective study is indicated to validate these findings. PMID- 10667511 TI - Initial severity of metabolic acidosis predicts the development of acute lung injury in severely traumatized patients. AB - OBJECTIVES: First, to determine whether the severity of shock, as measured by systemic hypotension and metabolic acidosis, is significantly associated with a higher risk of acute lung injury in patients with severe trauma. Second, to determine whether the volumes of blood and crystalloid solutions administered in the early posttrauma period are independent risk factors for acute lung injury in severely traumatized patients. DESIGN: Prospective observational study. SETTING: Level I urban trauma center in a university hospital. PATIENTS: A total of 102 severely injured, mechanically ventilated trauma patients with an Injury Severity Score > or =16 and aged between 18 and 75 yrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Initial clinical and laboratory data were collected in the emergency department, and on a daily basis thereafter during the patient's intensive care unit stay. Of the 102 severely injured patients enrolled, 42 developed acute lung injury (41%) and 60 did not (59%). A total of 93% of the trauma patients who developed acute lung injury during the 17-month study period were included in the study. Initial base deficit was significantly lower in patients who developed acute lung injury than in those who did not (-8.8+/-4.5 vs. -5.6+/-5.1, p<.01). The difference in systolic blood pressure between the two groups was not significant. CONCLUSIONS: In this group of severely injured trauma patients, the degree of metabolic acidosis at the time of admission identified those patients with the highest probability of developing acute lung injury. In addition, the volume of crystalloid solution administered during the first 24 hrs was significantly greater in patients who later developed acute lung injury. Finally, there was a significantly higher morbidity in patients who developed acute lung injury, whereas mortality did not differ between the two groups. PMID- 10667512 TI - Relationship between central venous pressure and bioimpedance vector analysis in critically ill patients. AB - OBJECTIVE: To assess the relationship between central venous pressure values and bioelectrical impedance vector analysis (BIVA), which may be used as complementary methods in the bedside monitoring of fluid status. DESIGN: Cross sectional evaluation of a consecutive sample. SETTING: Intensive care unit of a university hospital. PATIENTS: One hundred and twenty-one consecutive Caucasian, adult patients of either gender, for whom routine central venous pressure measurements were available. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Central venous pressure values and impedance vector components (i.e., resistance and reactance) were determined simultaneously. Total body water predictions were obtained from regression equations according to either conventional bioimpedance analysis or anthropometry (Watson and Hume formulas). Variability of total body water predictions was unacceptable for clinical purposes. Central venous pressure values significantly and inversely correlated with individual impedance vector components (r2 = .28 and r2 = .27 with resistance and reactance, respectively), and with both vector components together (R2 = .31). Patients were classified in three groups according to their central venous pressure value: low (0 to 3 mm Hg); medium (4 to 12 mm Hg); and high (13 to 20 mm Hg). Three BIVA patterns were considered: vectors within the target (reference) 75% tolerance ellipse (normal tissue hydration); long vectors out of the upper pole of the target (dehydration); and short vectors out of the lower pole of the target (fluid overload). The agreement between BIVA and central venous pressure indications was good in the high central venous pressure group (93% short vectors), moderate in the medium central venous pressure group (35% normal vectors), and poor in low central venous pressure group (10% long vectors). CONCLUSIONS: Central venous pressure values correlated with direct impedance measurements more than with total body water predictions. Whereas central venous pressure values >12 mm Hg were associated with shorter impedance vectors in 93% of patients, indicating fluid overload, central venous pressure values <3 mm Hg were associated with long impedance vectors in only 10% of patients, indicating tissue dehydration. The combined evaluation of intensive care unit patients by BIVA and central venous pressure may be useful in therapy planning, particularly in those with low central venous pressure in whom reduced, preserved, or increased tissue fluid content can be detected by BIVA. PMID- 10667513 TI - How much guidewire is too much? Direct measurement of the distance from subclavian and internal jugular vein access sites to the superior vena cava atrial junction during central venous catheter placement. AB - OBJECTIVE: The introduction of excessive lengths of guidewire during placement of central venous catheters from the internal jugular vein (IJV) or the subclavian vein (SCV) can result in rare but significant complications. To identify a "safe" guidewire insertion length, the authors performed direct intravascular measurement of the distance from these venous access sites to the superior vena cava-atrial junction (CAJ), and evaluated these distances relative to the patients' height, weight, sex, and chest radiographs. DESIGN: Prospective, nonrandomized observation. SETTING: The Interventional Radiology Department of a tertiary care referral hospital. PATIENTS: 100 adults (45 women, 55 men) evaluated during fluoroscopically directed central venous catheter placement. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The distance from the IJV or SCV access site was directly measured using fluoroscopy and an intravascular guidewire. 40 right IJVs, 31 right SCVs, 16 left SCVs, and 13 left IJVs were studied. Comparative measurements from the postprocedure radiograph were made in 20 of these cases. All measurements were correlated with patient sex, height, and weight. The mean distance from all access sites to the superior vena cava-atrial junction was 18.0 cm. The right IJV distance was the shortest, averaging 16 cm. The left SCV distance was the longest, averaging 21.2 cm. Right SCV and left IJV distances were 18.4 and 19.1 cm, respectively, but this difference was not statistically significant. Weight and radiographic measurements did not correlate with the measured vascular distance, although there was a trend toward longer distances in taller patients and males. CONCLUSIONS: Patient height, weight, and measurements from previous chest radiographs are less reliable in predicting a safe wire length than is the access site selected. In most cases, 18 cm should be considered the upper limit of guidewire introduced during central catheter placement in adults. The guidewires supplied in catheter kits should have lengths correlated to those of the catheters, and should have distance markings printed upon them. PMID- 10667514 TI - Protein carbonyl measurements show evidence of early oxidative stress in critically ill patients. AB - OBJECTIVE: To determine whether there is evidence of oxidative injury in patients who are critically ill with severe sepsis or major trauma, by measuring protein and lipid oxidation products. DESIGN: A prospective, observational study. SETTING: Critical care unit at a university teaching hospital. PATIENTS: Twenty two patients with severe sepsis (Acute Physiology and Chronic Health Evaluation II score 15-34) and eight patients with major trauma (Injury Severity Score 26 50). INTERVENTIONS: Plasma and bronchoalveolar lavage fluid was collected regularly during the first 10 days after trauma or onset of sepsis. Both fluids were analyzed for protein carbonyl concentrations as a measure of protein oxidation and thiobarbituric acid-reactive substances as a measure of lipid peroxidation. Myeloperoxidase concentrations were measured as an index of neutrophil activation. MEASUREMENTS AND MAIN RESULTS: Protein carbonyl concentrations were initially highly elevated compared with those in healthy adults in the plasma of both patient groups. They fell significantly within the first few days but remained above control values. Protein carbonyl concentrations were also high initially in bronchoalveolar lavage fluid and fell significantly with time. Thiobarbituric acid-reactive substances were not increased in plasma, and varied over a wide concentration range in lavage fluid. Myeloperoxidase activity reached micromolar levels in the lavage fluid when corrected for dilution, and was significantly higher in the plasma of the sepsis patients who subsequently died. There was a strong correlation between carbonyl concentrations in lavage fluid and plasma, and between protein carbonyls, thiobarbituric acid reactive substances and myeloperoxidase in the lungs. CONCLUSIONS: Our results provide evidence of oxidation occurring early in severe sepsis and major trauma patients, with protein carbonyl measurements providing a sensitive index of this process. High protein carbonyl concentrations in plasma as well as bronchial aspirates indicate that oxidation is not restricted to the lungs. The correlation between oxidative measures and myeloperoxidase concentrations in the lung indicates that neutrophil oxidants could be responsible for the injury. PMID- 10667515 TI - Short-term and long-term outcome prediction with the Acute Physiology and Chronic Health Evaluation II system after orthotopic liver transplantation. AB - OBJECTIVE: To evaluate the relationship between the postoperative Acute Physiology and Chronic Health Evaluation (APACHE) II score and mortality at hospital discharge and at 1 yr in liver transplant recipients. POPULATION: Adult orthotopic liver transplant (OLTX) recipients (n = 599) admitted to the intensive care unit postoperatively at a university hospital. METHODS: The cohort was split randomly into development and validation sets. Three models were compared for each end point: a) the original APACHE II slope with the original APACHE II postgastrointestinal surgery intercept; b) the original APACHE II slope with an OLTX-specific intercept generated from the development set; and c) an OLTX specific slope and intercept generated from the development set. Goodness-of-fit and calibration were assessed by the Hosmer-Lemeshow C statistic (where p>.05 suggests good fit) and standardized mortality ratios. Discrimination was assessed by receiver operator characteristic area under the curve analysis. MEASUREMENTS AND MAIN RESULTS: Hospital and 1-yr mortality rates were 9.9% and 15.9%, respectively. The APACHE II score was strongly associated with mortality (chi square, p<.0001), but when used with the original equation, it significantly overestimated hospital mortality (standardized mortality ratio, 0.73 [confidence interval, 0.58-0.99]). Using the OLTX-specific approaches, goodness-of-fit for both hospital and 1-yr mortality was good (p = .2-.57) but discrimination was only moderate (receiver operator characteristic area under the curve, 0.675 0.723). CONCLUSIONS: APACHE II is a good predictor of short- and long-term mortality after liver transplantation, especially when using OLTX-specific coefficients. Because fit and calibration were better than discrimination, APACHE II will be most useful in the prediction of risk for groups of patients (e.g., in clinical trials or institutional comparisons) rather than for individuals. This study raises the possibility that APACHE II may be useful for long-term mortality prediction in other critically ill populations. The overestimation of mortality using the original equation suggests that orthotopic liver transplantation, by reversing the underlying pathophysiology, may modify risk. PMID- 10667516 TI - Neonatal cerebral oxygen regulation after hypothermic cardiopulmonary bypass and circulatory arrest. AB - OBJECTIVE: Despite technical advances, neurologic sequelae continue to occur in neonates after heart surgery using deep hypothermic cardiopulmonary bypass (dhCPB) and circulatory arrest (DHCA). This study sought to determine the cerebral microcirculatory responses to hypoxia, hypotension, hypocapnia, and hypercapnia after dhCPB and DHCA. DESIGN: Prospective laboratory animal trial. SETTING: Research laboratory. SUBJECTS: Twenty-eight newborn pigs. INTERVENTIONS: Piglets were divided into control, dhCPB, and DHCA groups. The control group received surgery. The dhCPB group received surgery and deep hypothermic CPB for 40 mins. The DHCA group received surgery, deep hypothermic CPB for 40 mins, and circulatory arrest for 60 mins. Two hours after the intervention, cerebral microcirculatory responses were examined. MEASUREMENTS AND MAIN RESULTS: Cerebral microcirculatory responses consisted of changes in cerebral oxygen saturation (Sco2) and pial arteriolar diameter measured by near- infrared spectroscopy and intravital microscopy, respectively. All groups experienced similar decreases in Sco2 and increases in pial arteriolar diameter in response to moderate and severe hypoxia (Pao2, 35 and 25 torr, respectively) and moderate and severe hypotension (mean pressure, 30 and 20 mm Hg, respectively). Sco2 and pial arteriolar diameter decreased to hypocapnia (Paco2, 25 torr) similarly in all groups. To hypercapnia (Paco2, 70 torr), Sco2 increased in the control group, did not change in the dhCPB group, and decreased in the DHCA group. Pial arteriolar diameter to hypercapnia increased in the control and the dhCPB groups but did not change in the DHCA group. CONCLUSIONS: Cerebral vascular and oxygenation responses to hypoxia, hypocapnia, and hypotension were preserved after dhCPB and 1 hr of DHCA. By comparison, cerebral vascular and oxygenation responses to hypercapnia were not; both vascular and oxygenation responses were altered after DHCA, but only the oxygenation response was altered after dhCPB. These data suggest a selective disturbance in the microcirculation and/or parenchymal oxygen metabolism after DHCA and dhCPB. PMID- 10667517 TI - Hypertonic saline dextran produces early (8-12 hrs) fluid sparing in burn resuscitation: a 24-hr prospective, double-blind study in sheep. AB - OBJECTIVE: Resuscitation of large burn injuries must quickly restore and maintain cardiovascular function and fluid balance while minimizing secondary edema related damage. We tested the hypothesis that two 4-mL x kg(-1) doses of hypertonic saline dextran (HSD; 7.5% NaCl/6% dextran-70) can produce prolonged reduction in fluid requirements after burn injury. DESIGN: Prospective, pseudo randomized, double-blind study. SETTING: Animal research laboratory. SUBJECTS: Female adult Merino sheep (n = 12). INTERVENTIONS: Sheep were given a 40% total body surface area full-thickness flame burn under halothane anesthesia. One hour after the burn, the conscious animals received an initial dose of 4 mL x kg(-1) HSD (n = 6) or normal saline (NS; NaCl 0.9%) (n = 6) intravenously during 30 mins. This was followed by lactated Ringer's solution, infused to a target urine output of 1 mL x kg(-1) x hr(-1) throughout the 24-hr study. A second 4-mL x kg( 1) dose of HSD or NS was started at 12 hrs, and infused during 5 hrs. MEASUREMENTS AND MAIN RESULTS: Hourly urine output measurements were used to guide the infusion rate of the lactated Ringer's. The initial infusion of HSD 1 hr after the burn injury promptly restored cardiac index, promoted diuresis, and reduced fluid requirements compared with the NS controls (73% reduction for HSD relative to NS at 8 hrs). Subsequent rebound fluid accumulation resulted in similar net fluid balances in both groups within 12 hrs after the burn. The second dose of HSD, given at 12 hrs, was without effect on hemodynamics and fluid balance. CONCLUSIONS: We conclude a considerable initial, but not sustained fluid sparing effect of early HSD, and no effect of a late, slowly infused HSD dose in this two-dose regimen. PMID- 10667518 TI - Propofol impairment of mitochondrial respiration in isolated perfused guinea pig hearts determined by reflectance spectroscopy. AB - OBJECTIVE: To simultaneously determine the effect of propofol on myocardial oxygenation, mitochondrial function, and whole organ function in an isolated heart model, using optical reflectance spectroscopy. DESIGN: Controlled laboratory investigation. SETTING: Research laboratory. SUBJECTS: Twenty adult guinea pigs. INTERVENTIONS: Isolated hearts were perfused alternately with a modified oxygenated Krebs-Henseleit buffer and with buffer containing varied concentrations of propofol. Ninety seconds of ischemia were produced during perfusion with each solution studied. MEASUREMENTS AND MAIN RESULTS: Myoglobin oxygen saturation, cytochrome c and cytochrome a/a3 redox state, and ventricular pressure were continuously measured from isolated guinea pig hearts during a 2-hr period. Myoglobin oxygen saturation increased and both cytochromes became more oxidized in the presence of propofol. During ischemia, myoglobin desaturation and cytochrome reduction were delayed and less complete in the presence of propofol. The mean ischemic time to 50% myoglobin desaturation was, on average, 14.3 secs with buffer perfusion, and increased to 24.5, 27.9, and 41.8 secs, with 50, 100, and 200 microM propofol perfusion, respectively. Ventricular function decreased linearly with increasing propofol concentration. From baseline buffer perfusion, maximal dP/dt per cardiac cycle decreased on average by 30.4%, 40.9%, and 69.4%, with 50, 100, and 200 microM propofol perfusion, respectively. CONCLUSIONS: Propofol impairs either oxygen utilization or inhibits electron flow along the mitochondrial electron transport chain in the guinea pig cardiomyocyte. Propofol also significantly decreases ventricular performance in the isolated perfused heart. These effects are linearly correlated with propofol concentration in the range studied. PMID- 10667519 TI - Regional pressure volume curves by electrical impedance tomography in a model of acute lung injury. AB - OBJECTIVE: A new noninvasive method, electrical impedance tomography (EIT), was used to make pressure-impedance (PI) curves in a lung lavage model of acute lung injury in pigs. The lower inflection point (LIP) and the upper deflection point (UDP) were determined from these curves and from the traditional pressure-volume (PV) curves to determine whether the PI curves resemble the traditional PV curves. Furthermore, regional differences in the mentioned determinants were investigated. DESIGN: Prospective, experimental study. SETTING: Animal research laboratory. INTERVENTIONS: In nine anesthetized pigs, repeated lung lavage was performed until a Pao2 <80 torr was reached. Thereafter, an inspiratory PV curve was made using a constant flow of oxygen. During the intervention, EIT measurements were performed. MEASUREMENTS AND MAIN RESULTS: In this study, the LIP(EIT) was within 2 cm H2O of the LIP(PV). Furthermore, it was possible to visualize regional PI curves by EIT. No significant difference was found between the LIP(PV) (21.3+/-3.0 cm H2O) and the LIP(EIT) of the total lung (21.5+/-3.0 cm H2O) or the anterior parts of the lung (21.5+/-2.9 cm H2O). A significantly higher LIP (29.5+/-4.9 cm H2O) was found in the posterior parts of the lung. A UDP(PV) could be found in three animals only, whereas in all animals a UDP(EIT) could be determined from the anterior part of the lung. CONCLUSIONS: Using EIT, determination of LIP and UDP from the regional PI curves is possible. The obtained information from the regional PI curves may help in understanding alveolar recruitment. The use of this new bedside technique for clinical decision making remains to be examined. PMID- 10667520 TI - Effect of naloxone on immune responses after hemorrhagic shock. AB - OBJECTIVE: To determine whether naloxone administration after hemorrhagic shock has any beneficial or deleterious effect on immune responses. BACKGROUND DATA: Hemorrhagic shock is known to produce immunodepression in both humans and experimental animals. Although studies suggest that endogenous opioids play a role in immune regulation in adverse circulatory conditions, it remains controversial whether these opioids exert beneficial or detrimental effects on immunity after shock. Moreover, little information is available concerning the effects of opioid receptor blockade using naloxone on cell-mediated immunity and endocrine responses after shock. METHODS: Male C3H/HeN mice (25 g) were bled to and maintained at a mean arterial blood pressure of 35+/-5 mm Hg for 1 hr. The shed blood was then returned along with lactated Ringer's solution (two times the shed blood volume) to provide fluid resuscitation. The animals were randomized to receive either naloxone (1 mg/kg i.v.) or an equal volume of vehicle (saline) after the shed blood was returned, i.e., immediately before crystalloid resuscitation, and were killed at 2 hrs after resuscitation to obtain splenocytes, macrophages (peritoneal and splenic), and blood. MEASUREMENTS AND MAIN RESULTS: Bioassays revealed significantly decreased release of all studied interleukins (interleukins-1, -2, -3, and -6) by peritoneal and splenic macrophages as well as significantly decreased splenocyte proliferative capacity after shock in vehicle-treated mice. Naloxone administration after hemorrhage resulted in either similar or even more decreased levels of interleukin release compared with vehicle-treated hemorrhaged mice. Significantly increased plasma corticosterone concentrations were observed in vehicle-treated animals compared with control animals. Naloxone administration did not have any significant effects on corticosterone plasma concentrations after hemorrhage. CONCLUSIONS: These findings indicate the importance of the endogenous opioid system for the maintenance of immunity in adverse circulatory conditions, i.e., hemorrhage. Although additional studies involving different doses and/or times of naloxone administration may provide different results, the present findings raise the concern that naloxone administration in the traumatized host may have deleterious effects because it decreases peritoneal macrophage and splenic immune functions. PMID- 10667521 TI - Oxotremorine-induced cerebral hyperemia does not predict infarction volume in spontaneously hypertensive or stroke-prone rats. AB - OBJECTIVES: We tested the following hypotheses: a) spontaneously hypertensive stroke-prone rats (SHR-SP) have more brain injury than spontaneously hypertensive rats (SHR) and normotensive controls (Wistar-Kyoto rats [WKY]) when exposed to transient focal ischemia; b) infarction size is not correlated with baseline blood pressure; and c) infarction size is inversely related to the cerebral hyperemic response to oxotremorine, a muscarinic agonist that increases cerebral blood flow (CBF) by stimulating endothelial nitric oxide synthase. DESIGN: In vivo study. SETTING: Animal laboratory in a university teaching hospital. SUBJECTS: Adult age-matched male WKY, SHR, and SHR-SP. INTERVENTIONS: Rats were instrumented under halothane anesthesia. Transient focal cerebral ischemia was produced for 2 hrs with the intravascular suture technique. Cerebral perfusion, estimated with laser Doppler flowmetry (LD-CBF), in response to intravenous oxotremorine, was measured in one cohort of rats to estimate endothelial nitric oxide synthase function. Infarction volume was measured at 22 hrs of reperfusion with 2,3,5-triphenyltetrazolium chloride staining. MEASUREMENTS AND MAIN RESULTS: Infarction volume in the striatum of SHR-SP (42+/-4 mm3) was greater than in SHR (29+/-6 mm3) or WKY (1+/-1 mm3) (n = 9 rats/strain). Resting (unanesthetized) mean arterial blood pressure was similar in SHR-SP (177+/-5 mm Hg) and SHR (170+/ 5 mm Hg) despite a greater infarction volume in SHR-SP (n = 4) compared with SHR (n = 5). The percentage increase in LD-CBF signal in response to oxotremorine was similar for both groups (SHR, 64%+/-22% [n = 10]; SHR-SP, 69%+/-22% [n = 8]). However, in this cohort, cortical infarction volume was less in SHR (30%+/-4% of ipsilateral cortex) than in SHR-SP (49%+/-2% of ipsilateral cortex). CONCLUSIONS: Although SHR-SP have greater infarction volume than SHR, the mechanism of injury does not appear to be related to a difference in unanesthetized baseline mean arterial blood pressure or to an alteration in endothelium-produced nitric oxide. PMID- 10667522 TI - Antioxidant LY231617 enhances electrophysiologic recovery after global cerebral ischemia in dogs. AB - OBJECTIVE: The potent antioxidant LY231617 (2,6-bis(1,1-dimethylethyl)-4-[[(1 ethyl)amino]methyl]phenol hydrochloride) is cytoprotective in models of focal and global cerebral ischemia. We tested the hypothesis that administration of LY231617, before the insult, would improve recovery of cerebral electrical activity and metabolic function after transient global cerebral ischemia by improving cerebral blood flow (CBF) during the reperfusion period. DESIGN: Randomized, controlled, prospective study. SETTING: Research laboratory at a university teaching hospital. SUBJECTS: Twenty-four male beagle dogs. INTERVENTIONS: All experiments were performed under pentobarbital anesthesia and controlled conditions of normoxia, normocarbia, and normothermia. Twelve control dogs received 20 mL/kg saline (vehicle) bolus into the right atrium and 0.01 mL/kg/min i.v., beginning 20 mins before 13 mins of global cerebral ischemia (by aortic occlusion). The dogs in the drug-treated group received LY231617 as a 10 mg/kg bolus 20 mins before ischemia and 5 mg/kg/hr throughout reperfusion (n = 12). CBF was measured using radiolabeled microspheres. MEASUREMENTS AND MAIN RESULTS: Total CBF, cerebral oxygen consumption, and somatosensory evoked potentials (SEP) were measured during 240 mins of reperfusion. CBF was similar in both vehicle- and LY231617-treated animals at baseline and throughout the experimental period. In all animals, SEP became isoelectric between 60 and 100 secs after cross-clamping of the ascending aorta. SEP amplitude recovery was significantly higher in drug-treated animals compared with controls (73%+/-15% vs. 39%+/-14% [mean+/-SEM] from baseline at 120 mins [p<.05] and 86%+/-12% vs. 49%+/-14% from baseline at 240 mins [p< .05]). CONCLUSIONS: LY231617 improves recovery of cerebral electrical function after complete transient global ischemia via mechanisms unrelated to cerebral circulatory effects. PMID- 10667523 TI - Partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids. AB - OBJECTIVE: To determine the impact of partial liquid ventilation on the degree of pulmonary damage by reactive oxygen species in a model of acute lung injury caused by systemic endotoxemia. DESIGN: A prospective, controlled, in vivo, animal laboratory study. SETTING: Animal research facility of a health sciences university. SUBJECTS: Forty New Zealand White rabbits. INTERVENTIONS: Mature rabbits were anesthetized and instrumented with a tracheostomy and vascular catheters. Animals were assigned to receive either partial liquid ventilation (n = 16) with perflubron (18 mL/kg via endotracheal tube) or conventional mechanical ventilation (n = 16). Both groups were ventilated using similar strategies, with an Fio2 of 1.0 and tidal volume as required to obtain a normal Paco2. Animals were then given 0.9 mg/kg Escherichia coli endotoxin intravenously over 30 mins. Eight uninjured instrumented and mechanically ventilated animals served as controls. Partial liquid ventilation or conventional ventilation was continued for 4 hrs before the animals were killed. Lung homogenates were analyzed for malondialdehyde (MDA) and 4-hydroxy-2(E)-nonenal (4-HNE) concentrations using a colorimetric assay. To assess protein oxidative damage, carbonyl groups in protein side chains were derivatized with 2,4-dinitrophenylhydrazine followed by Western blotting with a dinitrophenylated-specific primary antibody. MEASUREMENTS AND MAIN RESULTS: MDA (713.42+/-662 vs. 1601.4+/-1156 nmol/g protein; p = .023) and MDA plus 4-HNE (1480.24+/-788 vs. 2675.2+/-1628 nmol/g protein; p = .038) concentrations were lower in animals treated with partial liquid ventilation compared with conventionally ventilated animals, respectively. Animals treated with partial liquid ventilation exhibited attenuation of dinitrophenylated derivatized protein bands by Western blotting, indicating a reduction in protein oxidative damage. The presence of perfluorocarbon did not interfere with the MDA assay when assessed by independent analysis in vitro. Perflubron did not serve as a sink for peroxyl radicals produced in the aqueous phase during separate in vitro oxidation experiments. CONCLUSIONS: Partial liquid ventilation attenuates oxidative damage to lipids and proteins during experimental acute lung injury. This finding is not caused by binding of lipid peroxidation products to perflubron or by the peroxyl radical scavenging properties of perflubron. PMID- 10667524 TI - Brain blood flow patterns after the development of congestive heart failure: effects of treadmill exercise. AB - OBJECTIVE: Congestive heart failure (CHF) is associated with left ventricular (LV) failure, neurohormonal system activation, and diminished exercise capacity. Although alterations in systemic vascular resistive properties have been recognized to occur with CHF, whether and to what degree perfusion abnormalities occur within the brain after the development of CHF remain poorly understood. Accordingly, the present study measured brain blood flow patterns in pigs after the development of pacing-induced CHF at rest and after treadmill-induced exercise. MEASUREMENTS AND MAIN RESULTS: Adult pigs (n = 6) were studied before and after the development of pacing-induced CHF (240 beats/min, 3 wks) at rest and with treadmill exercise (3 mph, 15 degrees incline, 10 mins). At rest, LV stroke volume was reduced nearly 45% with CHF compared with normal (20+/-2 vs. 36+/-3 mL; p<.05) and was associated with a more than four-fold increase in plasma catecholamines, renin activity, and endothelin concentration. At rest, global brain blood flow was reduced with CHF compared with the normal state (1.06+/-0.13 vs. 0.81+/-0.06 mL/min/g; p<.05). At rest, blood flow to the frontal lobe, cerebellum, and medullary regions was reduced by approximately 30% in the CHF group (p<.05). With treadmill exercise, LV stroke volume remained lower and neurohormonal concentrations remained higher in the pacing CHF state. Global brain blood flow increased significantly with treadmill exercise in both the normal and CHF states (4.58+/-1.36 and 2.01+/-0.29 mL/min/g; p<.05) but remained reduced in the CHF state compared with normal values (p<.05). In the CHF group, the relative increase in blood flow with exercise was significantly blunted in the parietal and occipital regions of the cerebrum and the suprapyramidal region of the medulla. CONCLUSIONS: The development of pacing-induced CHF was associated with diminished brain perfusion under resting conditions and with treadmill exercise. These perfusion abnormalities with pacing CHF were pronounced in specific regions of the brain. The defects in brain perfusion with the development of CHF may contribute to abnormalities in centrally mediated processes of cardiovascular regulation. PMID- 10667525 TI - Cerebrospinal fluid tumor necrosis factor-alpha, interleukin-1beta, interleukin 6, and interleukin-8 as diagnostic markers of cerebrospinal fluid infection in neurosurgical patients. AB - OBJECTIVE: To evaluate whether cerebrospinal fluid concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, or IL-8 may be used as diagnostic markers for the differential diagnosis of aseptic vs. bacterial meningitis and/or ventriculitis in neurosurgical patients. DESIGN: Prospective, observational study. SETTING: University teaching hospital. SUBJECTS: A total of 112 cerebrospinal fluid samples from 14 asymptomatic patients with normal cerebrospinal fluid after neurosurgery, 27 asymptomatic and 19 symptomatic patients with postneurosurgical aseptic meningitis, 32 patients with postneurosurgical cerebrospinal fluid infection, and 20 with severe subarachnoid and/or cerebral hemorrhage. MEASUREMENTS AND MAIN RESULTS: Specific ELISA kits were used to analyze TNF-alpha, IL-1beta, IL-6, and IL-8 concentrations on cerebrospinal fluid samples. Elevations in cerebrospinal fluid concentrations of TNF-alpha, IL-1beta, IL-6, and IL-8 were induced by different diseases or neurosurgical procedures, but cerebrospinal fluid bacterial infection induced the highest concentrations. To discriminate between aseptic cerebrospinal fluid pleocytosis and cerebrospinal fluid infection with a specificity of 95%, cerebrospinal fluid leukocyte count >1700/mL, TNF-alpha >150 pg/mL, and IL-1beta >90 pg/mL showed sensitivities of 51%, 74%, and 90%, respectively. Sufficiently sensitive and specific cutoff points could not be found for cerebrospinal fluid IL-6 or IL-8. CONCLUSION: Cerebrospinal fluid IL-1beta appears to be the best biochemical marker of cerebrospinal fluid infection in neurosurgical patients. PMID- 10667526 TI - Post-intensive care unit pediatric hospital stay and estimated costs. AB - OBJECTIVE: For pediatric intensive care unit (ICU) survivors, to determine the proportion of hospital stay and estimated hospital costs accounted for by post ICU care. DESIGN: Prospective study. SETTING: University teaching hospital. PATIENTS: Pediatric patients who survive an ICU admission. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Estimated relative daily costs were assumed as follows: ICU, with ventilator/ICU, not on ventilator/intermediate care unit/general pediatric hospital day, at 2:1:0.7:0.3, respectively. Estimated costs were expressed in arbitrary units as (number of days at each level of care) x (relative cost per day). The ICU phase was defined as the patient's first ICU admission only, and the post-ICU phase included intermediate care unit and general pediatric hospital days, as well as ICU readmission during the same hospitalization. Pre-ICU hospital activity was excluded from analysis. For 341 ICU survivors, post-ICU days (median, 4 days per patient) accounted for 62% of the total hospital stay. Post-ICU care accounted for one third of the total estimated hospital costs for ICU survivors. Patients with longer post-ICU stays could not be reliably identified at the time of ICU discharge according to their ICU length of stay, duration of mechanical ventilation in the ICU, age, ICU day 1 mortality probability, or diagnostic group (p>.05). CONCLUSIONS: Post-ICU care accounts for a substantial proportion of hospital stay and estimated costs for ICU survivors. These observations suggest that developing strategies to optimize hospital utilization at the time of ICU discharge may be important for controlling costs of recovery from critical illness. PMID- 10667527 TI - Capillary blood gases in a pediatric intensive care unit. AB - OBJECTIVE: To determine if samples obtained from arterial and capillary sources are comparable in children with diverse pathologic conditions during their stay in a pediatric intensive care unit. STUDY DESIGN: Prospective, descriptive study in patients admitted to a multidisciplinary pediatric intensive care unit. INTERVENTIONS: Seventy-five simultaneous paired samples (arterial and capillary) were obtained from patients with different degrees of capillary reperfusion, hemodynamic stability, blood pressure, and temperature. Both samples were analyzed < or =5 mins after collection. MEASUREMENTS AND MAIN RESULTS: The average correlations between arterial and capillary samples were 0.87 for pH, 0.86 for CO2, and 0.65 for oxygen. Neither poor perfusion nor low temperature altered the correlation for pH or CO2. The only condition that significantly affected the correlation was hypotension. CONCLUSION: Capillary blood sampling is a useful alternative to gasometric evaluation of critically ill children, even in the presence of hypothermia or hypoperfusion, provided that hypotension is not present. PMID- 10667528 TI - Conventional ventilation versus high-frequency oscillation: hemodynamic effects in newborn babies. AB - OBJECTIVE: We conducted a prospective study to assess the hemodynamic effects of conventional mechanical ventilation (CMV) compared with high-frequency oscillation (HFO) in newborn babies with respiratory distress syndrome. METHODS: A total of 18 consecutive term and preterm infants were examined by two dimensional M-mode and pulsed Doppler echocardiography. RESULTS: Five patients had to be excluded, three of them because of increasing cardiovascular support after initiation of HFO. The remaining 13 infants (seven males, six females) had a median gestational age of 33 wks (range, 25-40) and a birth weight of 2350 g (range, 790-3600). Patients entered the study at 21 hrs (range, 5-69) of life, receiving total maintenance fluid of 90 mL/kg/day (range, 60-120). Five babies (38%) needed continuous inotropic support. HFO was used as a rescue therapy in infants who failed with CMV. In all 13 patients, HFO significantly impaired cardiac performance compared with CMV by decreasing aortic velocity-time integral: median, 10.2 cm (range, 6.0-14.6) vs. 8.3 cm (range, 5.3-12.4; p<.002); stroke volume: median, 3.8 mL (range, 1.6-6.8) vs. 3.2 mL (range, 1.3-5.9; p<.002); and cardiac index: 281 mL/min/kg of body weight (range, 177-579) vs. 200 mL/min/kg of body weight (range, 156-591; p<.002). Fractional shortening was also significantly reduced: median, 0.31% (range, 0.24-0.44) vs. 0.29% (range, 0.20 0.34; p<.01), because of a significantly smaller left ventricular diastolic diameter during HFO: median, 1.4 cm (range, 1.0-1.9) vs. 1.4 cm (range, 0.9-1.8; p<.05), with a median difference of -0.07 cm (range, -0.4-0.2). HFO also causes a significant decrease in heart rate-corrected left ventricular ejection time: median, 0.25 sec (range, 0.23-0.28) vs. 0.23 sec (range, 0.21-0.26; p < .02) and heart rate-corrected velocity of circumferential fiber shortening (Vcfc): median, 1.3 circ/sec (range, 1.0-1.6) vs. 1.2 circ/sec (range, 0.9-1.4; p<.05). Left ventricular end-systolic wall stress (LVESWS; g/cm2) remained stable. The correlation between Vcfc and LVESWS did not show any significance (CMV, r2 = .2; HFO, r2 = .09). The regression line between Vcfc and LVESWS showed a higher y intercept and steeper slope during CMV than during HFO. Heart rate, mean arterial pressure, and left ventricular systolic diameter remained unchanged. CONCLUSIONS: In newborn babies, HFO significantly decreased left ventricular cardiac output caused by reduced left ventricular filling and HFO decreased contractility at higher mean airway pressures than with CMV. PMID- 10667529 TI - Consensus conference definitions for sepsis, septic shock, acute lung injury, and acute respiratory distress syndrome: time for a reevaluation. AB - Definitions for sepsis, septic shock, acute lung injury (ALI), and acute respiratory distress syndrome (ARDS) were developed by consensus conferences with the goal of achieving standardization of terminology and improved homogeneity of patient populations in clinical studies. Although such definitions have been useful in epidemiologic investigations, the criteria specified by the consensus conferences are broad and insufficiently specific to address the problem of heterogeneous mechanisms leading to clinical syndromes. An important challenge is to progress from clinical syndromes, as presently defined, to more specific entities that are delineated by alterations in specific immunologic or biochemical pathways. Such mechanistic definitions will provide more homogeneous groups of patients who can be identified at early stages of their clinical course. This approach encourages focused investigation of pathways leading to organ system dysfunction and death and, also, provides an efficient framework for the development of new therapies useful in critically ill patients. PMID- 10667530 TI - Consensus report for regionalization of services for critically ill or injured children. Council of the Society of Critical Care Medicine. AB - The care of critically ill children has become more complex and demanding. This document establishes recommendations for developing regionalized integration of the care of these children into the emergency medical services system. These recommendations were developed by pediatricians with expertise in pediatric critical care, transport, and emergency medicine from the Pediatric Section of the Society of Critical Care Medicine Task Force on Regionalization of Pediatric Critical Care and the Committee on Pediatric Emergency Medicine from the American Academy of Pediatrics. The document was developed from existing guidelines from a number of professional organizations (including the Society of Critical Care Medicine and the American Academy of Pediatrics), a thorough review of the literature, and expert consensus. PMID- 10667531 TI - Unexpected Ebola virus in a tertiary setting: clinical and epidemiologic aspects. AB - OBJECTIVES: To describe the clinical manifestations of viral hemorrhagic fever, and to increase clinicians' awareness and knowledge of these illnesses. DESIGN: Retrospective study of the clinical and laboratory data and management of two cases of Ebola virus infection with key epidemiologic data provided. SETTING: Two tertiary care hospitals. PATIENTS: Two adult patients, the index case and the source patient, both identified as having Ebola, one of whom originated in Gabon. INTERVENTIONS: One patient was admitted to the intensive care unit. The other was managed in a general ward. MEASUREMENT AND MAIN RESULTS: Clinical and laboratory data are reported. One patient, a healthcare worker who contracted this illness in the course of her work, died of refractory thrombocytopenia and an intracerebral bleed. The source patient survived. Despite a long period during which the diagnosis was obscure, none of the other 300 contacts contracted the illness. CONCLUSIONS: Identification of high-risk patients and use of universal blood and body fluid precautions will considerably decrease the risk of nosocomial spread of viral hemorrhagic fevers. PMID- 10667532 TI - Use of high-dose corticosteroids and high-frequency oscillatory ventilation for treatment of a child with diffuse alveolar hemorrhage after bone marrow transplantation: case report and review of the literature. AB - BACKGROUND: Other than relapse, pulmonary complications are the most common cause of mortality in patients who undergo bone marrow transplantation (BMT). Diffuse alveolar hemorrhage (DAH) is one noninfectious pulmonary complication of BMT. Presenting clinical findings include nonproductive cough usually without hemoptysis, dyspnea, hypoxemia, a decrease in hematocrit, and diffuse infiltrates on chest radiograph. PATIENT: We report a case of DAH after allogeneic BMT in a 6 yr-old female patient. Although a chest radiograph revealed patchy bilateral alveolar densities and large volumes of bright red blood were suctioned from the endotracheal tube, there was no evidence of coagulopathy and no infectious agent was identified on examination of bronchoalveolar lavage fluid, blood, and urine. INTERVENTION: The child was treated with high-dose corticosteroids and high frequency oscillatory ventilation and experienced a complete clinical recovery from her pulmonary disease. RESULTS: The definition, presenting symptoms, findings and timing, and associated risk factors of DAH after BMT are reviewed. Prospective hypotheses for the pathogenesis of DAH after BMT are presented. Evidence for the role of high-dose corticosteroids for treatment of DAH after BMT and the role of high-frequency oscillatory ventilation for treatment of acute hypoxemic respiratory failure in children with diffuse alveolar disease is also reviewed. CONCLUSION: This case supports the contention that early treatment with high-dose corticosteroids is warranted in children with DAH after BMT. PMID- 10667533 TI - Vasopressin as an alternative to norepinephrine in the treatment of milrinone induced hypotension. AB - OBJECTIVE: To determine whether vasopressin could be effective in treating the hypotension associated with phosphodiesterase III inhibition. Phosphodiesterase III inhibitors are cardiotonic agents that increase myocardial contractility and decrease vascular smooth muscle tone. The vasodilatory effect can be profound, and the resulting hypotension frequently requires the administration of catecholamine pressors. DESIGN: Retrospective analysis of existing data. SETTING: The medical or surgical intensive care unit of Columbia-Presbyterian Medical Center. PATIENTS: Three consecutive patients receiving milrinone and requiring catecholamine pressors to maintain systolic arterial pressure of > or =90 mm Hg. INTERVENTIONS: Vasopressin was administered to the three patients. MEASUREMENTS AND MAIN RESULTS: Vasopressin (0.03-0.07 units/min) increased systolic arterial pressure from 90+/-4.7 to 130+/-2.3 mm Hg while reducing the administration of catecholamine pressors. CONCLUSIONS: Vasopressin at very low doses appears to be an effective vasopressor for milrinone-induced hypotension. PMID- 10667534 TI - What role does neutrophil apoptosis play in acute respiratory distress syndrome? PMID- 10667535 TI - Use of immune globulin intravenous (human) to prevent infection in the multiple trauma patient. PMID- 10667536 TI - Science and surveys in the treatment of head injuries. PMID- 10667537 TI - Outcomes of post-intensive care unit care: once more, the half-full/empty glass. PMID- 10667538 TI - Prognostic systems in intensive care: how do you interpret an observed mortality that is higher than expected? PMID- 10667539 TI - Acute quadriplegia myopathy in the intensive care unit: time to look at a mechanistic approach. PMID- 10667540 TI - Upper gastrointestinal hemorrhage in the inpatient hospital setting: a different beast? PMID- 10667541 TI - Leukotriene B4, acute respiratory distress syndrome, and outcomes. PMID- 10667542 TI - Are resuscitation fluids harmful? PMID- 10667543 TI - Transport of the critically ill. PMID- 10667544 TI - Ionized hypocalcemia during sepsis. PMID- 10667545 TI - Outcomes research at the end of life. PMID- 10667546 TI - Redox pairs, tissue hypoxia, organ dysfunction, and mortality. PMID- 10667547 TI - Scoring systems for eclampsia. PMID- 10667548 TI - Bioelectrical impedance monitoring: is this a bull market for electric utilities? PMID- 10667549 TI - Quantifying the rancid, the rotten, and the rusty related to oxidant-mediated molecular pathogenesis. PMID- 10667550 TI - Does hypertonic burn resuscitation make a difference? PMID- 10667551 TI - Naloxone and shock. PMID- 10667552 TI - A complex role for nitric oxide in ischemic stroke. PMID- 10667553 TI - Aseptic versus bacterial postoperative meningitis: cytokines as a distinguishing marker. PMID- 10667554 TI - Hemodynamic effects of high-frequency oscillatory ventilation: a little volume goes a long way. PMID- 10667555 TI - An outbreak of Ebola virus: lessons for everyday activities in the intensive care unit. PMID- 10667556 TI - Effects of intra-aortic balloon occlusion during cardiopulmonary resuscitation. PMID- 10667557 TI - Use of animal models to test novel therapies in sepsis. PMID- 10667558 TI - Gastric pH and intravenous ranitidine in critically ill children. PMID- 10667559 TI - The relatives' place at resuscitation. PMID- 10667560 TI - Regulation of angiogenesis via vascular endothelial growth factor receptors. PMID- 10667561 TI - Malonyl-coenzyme-A is a potential mediator of cytotoxicity induced by fatty-acid synthase inhibition in human breast cancer cells and xenografts. AB - A biologically aggressive subset of human breast cancers and other malignancies is characterized by elevated fatty-acid synthase (FAS) enzyme expression, elevated fatty acid (FA) synthesis, and selective sensitivity to pharmacological inhibition of FAS activity by cerulenin or the novel compound C75. In this study, inhibition of FA synthesis at the physiologically regulated step of carboxylation of acetyl-CoA to malonyl-CoA by 5-(tetradecyloxy)-2-furoic acid (TOFA) was not cytotoxic to breast cancer cells in clonogenic assays. FAS inhibitors induced a rapid increase in intracellular malonyl-CoA to several fold above control levels, whereas TOFA reduced intracellular malonyl-CoA by 60%. Simultaneous exposure of breast cancer cells to TOFA and an FAS inhibitor resulted in significantly reduced cytotoxicity and apoptosis. Subcutaneous xenografts of MCF7 breast cancer cells in nude mice treated with C75 showed FA synthesis inhibition, apoptosis, and inhibition of tumor growth to less than 1/8 of control volumes, without comparable toxicity in normal tissues. The data suggest that differences in intermediary metabolism render tumor cells susceptible to toxic fluxes in malonyl CoA, both in vitro and in vivo. PMID- 10667562 TI - Redox regulation of glutathione S-transferase induction by benzyl isothiocyanate: correlation of enzyme induction with the formation of reactive oxygen intermediates. AB - Here we report the molecular mechanism underlying the induction of glutathione S transferase (GST) in rat liver epithelial RL34 cells treated with a cancer chemopreventive isothiocyanate compound, benzylisothiocyanate (BITC). BITC was found to significantly induce GST activity in RL34 cells. Northern and Western blot analyses demonstrated that BITC specifically enhanced the production of the class pi GST isozyme (GSTP1). Our studies demonstrated for the first time that the addition of BITC to the cells resulted in an immediate increase in the reactive oxygen intermediates (ROIs) detected by a fluorescence probe, 2',7' dichlorofluorescin diacetate. The level of the ROIs in the cells treated with BITC (10 microM) was approximately 50-fold higher than those in the control cells. Furthermore, glutathione depletion by diethyl maleate significantly enhanced BITC-induced ROI production and accelerated the BITC-induced elevation of the GST activity, whereas pretreatment of the cells with glutathione inhibited both the ROI production and GST induction. The structure-activity relationship of the isothiocyanates also indicated that the ROI-producing activities closely correlated with their GST-inducing potencies. Moreover, the GSTP1 enhancer I containing region was found to be essential for induction of the GSTP1 gene by intracellular ROI inducers such as BITC and diethyl maleate. These data suggest the involvement of the redox regulation on the induction of GSTP1 by BITC. PMID- 10667563 TI - Evidence that the epidermal targets of carcinogen action are found in the interfollicular epidermis of infundibulum as well as in the hair follicles. AB - Actively cycling, transit-amplifying cells and quiescent cells including stem cells are found in the layer of the epidermis and hair follicles. To determine the origin of skin tumors, we completely removed the interfollicular epidermis of carcinogen-initiated mice by an abrasion technique known to leave the hair follicles undisturbed. The interfollicular epidermis of the abraded mice quickly regenerated from cells in the hair follicles, after which time tumor promotion was begun. Mice in which the interfollicular epidermis had been removed developed papillomas and carcinomas; however, the number of papillomas throughout 40 weeks was half that of the unabraded mice. Carcinoma responses were not significantly different in the abraded and unabraded groups. These results are consistent with the hypothesis that the targets of tumor initiation are stem cells found in the hair follicles and, to a lesser degree, in the interfollicular epidermis. PMID- 10667564 TI - Vascular protection by chloroquine during brain tumor therapy with Tf-CRM107. AB - Tf-CRM107 is a conjugate of transferrin and a point mutant of diphtheria toxin that selectively kills cells expressing high levels of the transferrin receptor. Tf-CRM107 has been infused intratumorally into patients with malignant brain tumors. Although approximately half of the patients exhibit tumor responses, patients receiving higher doses of Tf-CRM107 may develop magnetic resonance image (MRI) evidence of toxicity indicative of small vessel thrombosis or petechial hemorrhage. Consistent with these clinical results we found that intracerebral injection of Tf-CRM107 into rats at total doses > or =0.025 microg causes brain damage detectable by MRI and histology. To widen the therapeutic window of Tf CRM107, we explored ways to prevent this damage to the vasculature. We reasoned that the vasculature may be protected to a greater extent than tumor from Tf CRM107 infused into brain parenchyma by i.v. injection of reagents with low blood brain barrier permeability that block the toxicity of Tf-CRM107. Chloroquine, a well-characterized antimalarial drug, blocks the toxicity of diphtheria toxin and Tf-CRM107. Systemic administration of chloroquine blocked the toxicity of Tf CRM107 infused intracerebrally in rats and changed the maximum tolerated dose of Tf-CRM107 from 0.2 to 0.3 microg. Moreover, chloroquine treatment completely blocked the brain damage detected by MRI caused by intracerebral infusion of 0.05 microg of Tf-CRM107. In nude mice bearing s.c. U251 gliomas, chloroquine treatment had little effect on the antitumor efficacy of Tf-CRM107. Thus, chloroquine treatment may be useful to reduce the toxicity of Tf-CRM107 for normal brain without inhibiting antitumor efficacy and increase the therapeutic window of Tf-CRM107 for brain tumor therapy. PMID- 10667565 TI - Induction of uterine adenocarcinoma in CD-1 mice by catechol estrogens. AB - Catechol estrogens may mediate estrogen-induced carcinogenesis because 4 hydroxyestradiol induces DNA damage and renal tumors in hamsters, and this metabolite is formed in the kidney and estrogen target tissues by a specific estrogen 4-hydroxylase. We examined the carcinogenic potential of catechol estrogen in an experimental model previously reported to result in a high incidence of uterine adenocarcinoma after neonatal exposure to diethylstilbestrol. Outbred female CD-1 mice were treated with 2- or 4 hydroxyestradiol, 17beta-estradiol, or 17alpha-ethinyl estradiol on days 1-5 of neonatal life (2 microg/pup/day) and sacrificed at 12 or 18 months of age. Mice treated with 17beta-estradiol or 17a-ethinyl estradiol had a total uterine tumor incidence of 7% or 43%, respectively. 2-Hydroxyestradiol induced tumors in 12% of the mice, but 4-hydroxyestradiol was the most carcinogenic estrogen, with a 66% incidence of uterine adenocarcinoma. Both 2- and 4-hydroxylated catechols were estrogenic and increased uterine wet weights in these neonates. These data demonstrate that both 2- and 4-hydroxyestradiol are carcinogenic metabolites. The high tumor incidence induced by 4-hydroxyestradiol supports the postulated role of this metabolite in hormone-associated cancers. PMID- 10667566 TI - Nickel compounds are novel inhibitors of histone H4 acetylation. AB - Environmental factors influence carcinogenesis by interfering with a variety of cellular targets. Carcinogenic nickel compounds, although generally inactive in most gene mutation assays, induce chromosomal damage in heterochromatic regions and cause silencing of reporter genes when they are located near telomere or heterochromatin in either yeast or mammalian cells. We studied the effects of nickel on the lysine acetylation status of the NH2-terminal region of histone H4. At nontoxic levels, nickel decreased the levels of histone H4 acetylation in vivo in both yeast and mammalian cells, affecting only lysine 12 in mammalian cells and all of the four lysine residues in yeast. In yeast, lysine 12 and 16 were more greatly affected than lysine 5 and 8. Interestingly, a histidine Ni2+ anchoring site is found at position 18 from the NH2-terminal tail of H4. Nickel was also found to inhibit the acetylation of H4 in vitro using purified recombinant histone acetyltransferase. To our knowledge, this is the first agent shown to decrease histone H4 acetylation at nontoxic levels. PMID- 10667567 TI - Prognostic significance of cyclin E overexpression in resected non-small cell lung cancer. AB - Cyclin E plays a pivotal role in the regulation of G1-S transition and relates to malignant transformation of the cells. However, the clinical significance of cyclin E expression in patients with non-small cell lung cancer remains unknown. We examined the expression of cyclin E in 242 resected non-small cell lung cancer in pathological stages I-IIIa and analyzed its relation to clinicopathological factors. Cylin E overexpressions were observed frequently in deeply invasive tumors. Multivariate analysis revealed that complete resection, pathological stage, and cyclin E expression were independent prognostic indicators. When cyclin E and proliferating cell nuclear antigen are combined, the cases negative for both had a significantly better prognosis than the other cases. We concluded that cyclin E overexpression relates to deeply invasive tumors and is correlated with poor prognosis. New therapeutic options may be provided by combination of cyclin E expression and proliferating cell nuclear antigen overexpression. PMID- 10667568 TI - Selective loss of estrogen receptor beta in malignant human colon. AB - Epidemiological data suggest a protective effect for estrogen replacement therapy on colon cancer. The estrogen receptor (ER) is required for the action of estrogen. The ER-beta isoform is functionally similar to ER-alpha but has a distinct pattern of expression and transcriptional response to selective estrogen response modulators. Our goal was to investigate the presence of ER-alpha and ER beta in normal and malignant colon tissue. Human colon cancer tissue and adjacent normal colon tissue were harvested from five male and six female patients undergoing segmental colon resection for colon cancer. Western blot analysis revealed very low levels of ER-alpha protein in tumor and normal colon tissue. In both male and female patients, malignant colon tissue showed a selective loss of ER-beta protein expression when compared to normal colon tissue in the same patient. Semiquantitative reverse transcription-PCR revealed no difference in ER beta mRNA levels between normal and malignant colon tissue. Malignant transformation of the colon is associated with a marked diminution of ER-beta protein expression, possibly through a posttranscriptional mechanism. PMID- 10667569 TI - Effects of cyclin D1 polymorphism on age of onset of hereditary nonpolyposis colorectal cancer. AB - A common polymorphism in the cyclin D1 gene enhances the gene's alternate splicing. The alternatively spliced product encodes an altered protein that does not contain sequences involved in the turnover of the protein. We found that hereditary nonpolyposis colorectal carcinoma patients who were homozygous or heterozygous for the mutant allele developed colorectal cancer an average of 11 years earlier than patients who were homozygous for the normal alleles. This is the first report indicating that the cyclin D1 polymorphism influences age of onset of cancer. Because cyclin D1 plays an important role in the G1 to S phase transition of the cell cycle, our findings suggest that cells with the mutant allele accumulate mutations as a result of defective mismatch repair and may also bypass the G1-S checkpoint of the cell cycle more easily than in cells not carrying the polymorphism. The polymorphism has a dominant phenotype. PMID- 10667571 TI - The HRAS1 minisatellite locus and risk of ovarian cancer. AB - Approximately 10% of ovarian cancers are due to mutations in highly penetrant inherited cancer susceptibility genes. The highly polymorphic HRAS1 minisatellite locus, located just downstream from the proto-oncogene H-ras-1 on chromosome 11p, consists of four common progenitor alleles and several dozen rare alleles, which apparently derive from mutations of the progenitors. Mutant alleles of this locus represent a major risk factor for cancers of the breast, colorectum, and bladder, and it was found that BRCAI mutation carriers with at least one rare HRAS1 allele have a greater risk of ovarian cancer than BRCA1 carriers with only common HRAS1 alleles. There are no conclusive studies of HRAS1 alleles in sporadic epithelial ovarian cancer. A case-control study of HRAS1 alleles was performed on DNA from 136 Caucasian patients with ovarian cancer and 108 cancer-free controls using conventional (Southern blot) and PCR-based methods to determine the frequency of rare HRAS1 alleles. Odds ratios (ORs) were estimated using unconditional logistic regression methods. A single degree of freedom test was used to assess the significance of linear trend across categories of increasing exposure. A statistically significant association between rare HRAS1 alleles and risk of ovarian cancer was observed [OR, 1.70; 95% confidence interval (CI), 1.03-2.80; P = 0.04]. Having only one rare allele was associated with a relative risk of 1.66 (95% CI, 0.91-3.01), whereas having two rare alleles increased the relative risk to 2.86 (95% CI, 0.75-10.94; trend P = 0.03). Analysis of HRAS1 allele types by the age of the case at diagnosis revealed that younger cases (<45 years) had a borderline statistically significant increased association with rare HRAS1 alleles compared to older cases (> or = 0 years; OR, 1.89; 95% CI, 0.90-3.98; P = 0.09). Rare HRAS1 alleles contribute to ovarian cancer predisposition in the general population. Thus, the HRAS1-variable number of tandem repeats locus may function as a modifier of ovarian cancer risk in both sporadic and hereditary ovarian cancer. PMID- 10667570 TI - Genetic vaccination with "self" tyrosinase-related protein 2 causes melanoma eradication but not vitiligo. AB - "Self" melanocyte differentiation antigens are potential targets for specific melanoma immunotherapy. Vaccination against murine tyrosinase-related protein (TRP)-1/gp75 was shown recently to cause melanoma rejection, which was accompanied by autoimmune skin depigmentation (vitiligo). To further explore the linkage between immunotherapy and autoimmunity, we studied the response to vaccination with a related antigen, TRP-2. i.m. inoculation of plasmid DNA encoding murine trp-2 elicited antigen-specific CTLs that recognized the B16 mouse melanoma and protected the mice from challenge with tumor cells. Furthermore, mice bearing established s.c. B16 melanomas rejected the tumor upon vaccination with a recombinant vaccinia virus encoding trp-2. Depletion experiments showed that CD8+ lymphocytes and natural killer cells were crucial for the antitumor activity of the trp-2-encoding vaccines. Mice that rejected the tumor did not develop generalized vitiligo, indicating that protective immunity can be achieved in the absence of widespread autoimmune aggression. PMID- 10667573 TI - Prostate mutations in rats induced by the suspected human carcinogen 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine. AB - Male lacl transgenic rats were fed a diet containing 200 ppm of 2-amino-1-methyl 6-phenylimidazo[4,5-b]pyridine (PhIP), a heterocyclic amine present in cooked meats. PhIP was found to be a powerful prostate mutagen. After 61 days of treatment, the lacl mutant frequency was 71 x 10(-5), >20-fold higher than the spontaneous mutant frequency of 3.2 x 10(-5). The predominant PhIP-induced mutations were G:C->T:A transversions and deletions of G:C bp. The results directly link PhIP-induced mutations with the earlier observation of PhIP-induced prostate cancer in rats and suggest that exposure to dietary PhIP could be a risk factor in the incidence of human prostate cancer. PMID- 10667572 TI - Mechanism for inactivation of the KIP family cyclin-dependent kinase inhibitor genes in gastric cancer cells. AB - The mechanism for inactivation of the KIP family cyclin-dependent kinase inhibitor (CDKI) genes, the p21, p27, and p57 genes, in gastric cancer cells was tested by treating the cells with either the DNA demethylation agent, 5-aza-2' deoxycytidine or the histone deacetylase inhibitor, n-butyric acid or trichostatin A. RNA expression of the gene was determined by reverse transcription PCR. The p21 gene was activated only by histone deacetylase inhibitor. The p57 gene was activated by histone deacetylase inhibitors in all of the gastric cancer cell lines and by 5-aza-2'-deoxycytidine in five of eight gastric cell lines. However, the p27 gene was not inactivated in gastric cancer cell lines. The methylation status of the promoter of the p21 and p57 genes was also tested by digestion with the methylation-sensitive restriction enzymes and a subsequent PCR. The promoter of the p21 gene has no methylation. The promoter of the p57 gene is, however, methylated in five of eight gastric cancer cell lines as expected from the result of the treatment with 5-aza-2'-deoxycytidine. Formation of the inactive chromatin through histone deacetylation seems to be the general mechanism for inactivation of both the p21 and the p57 genes in gastric cancer cells. Hypermethylation of promoter region seems to be an alternative pathway for inactivation of the p57 gene. PMID- 10667574 TI - The angiogenic factor interleukin 8 is induced in non-small cell lung cancer/pulmonary fibroblast cocultures. AB - The interactions between tumor cells and surrounding stromal elements may promote the release of angiogenic factors. Although interleukin 8 (IL-8) is a major angiogenic factor in non-small cell lung cancer (NSCLC), the stromal contribution to IL-8 expression in primary NSCLC remains to be defined. To elucidate the role of stromal elements in NSCLC IL-8 production, normal pulmonary fibroblasts were cocultured with six representative NSCLC lines in direct and transwell assays. IL 8 transcripts and protein were consistently induced in fibroblasts and a subset of NSCLCs as a consequence of tumor/stromal coculture. In these cocultures, IL-8 was induced by IL-1alpha and an additional, as yet unidentified, soluble factor. These data underscore the importance of tumor/stromal interaction in the production of angiogenic peptides such as IL-8 in NSCLC. PMID- 10667575 TI - Tamoxifen chemoprevention of a hormone-independent tumor in the proto-neu transgenic mice model. AB - We evaluated the effect of tamoxifen (TAM) on tumor development in proto-neu transgenic mice that spontaneously develop mammary carcinomas overexpressing the neu protein. These mammary carcinomas are hormone independent because superimposable growth of transplants was observed in females and males. Virgin transgenic mice treated with TAM from 24 weeks of age, ie., when subclinical mammary tumors are already present, showed a slightly accelerated tumor development. In contrast, transgenic mice treated with TAM starting at 12 weeks of age, when subclinical tumors are not yet present, showed a 50% reduction of tumor incidence. Light microscopy analysis of the mammary gland of these mice revealed an apparently normal ductal branching but a complete regression of the acini. In conclusion, TAM can prevent the occurrence of hormone-independent breast carcinoma if given early enough to inhibit normal cells. PMID- 10667576 TI - Heregulin induces expression, DNA binding activity, and transactivating functions of basic leucine zipper activating transcription factor 4. AB - Heregulin beta1 (HRG), a combinatorial ligand for human epidermal growth factor receptor 3 and human epidermal growth factor receptor 4 receptors, is a regulatory secretory polypeptide with distinct biological effects such as growth stimulation, differentiation, invasiveness, and migration in breast cancer cells. The mechanism underlying the diverse functions of HRG is not well established, but it is believed to be dependent on the induced changes in expression of specific cellular gene products, their modification, or both. The binding of basic leucine zipper transcription factors to the cAMP response element is known to activate a variety of gene products with a role or roles in growth regulation. In the studies presented here, we identified basic leucine zipper activating transcription factor (ATF) 4 as one of the HRG-inducible gene product. We demonstrated that HRG stimulation of human cancer cells induces expression of ATF4 mRNA and protein, ATF4 DNA binding activity, and ATF4 transactivating function. Consistent with its role as a transcriptional activator, HRG-stimulated ATF4 protein stimulated the transcription from an artificial promoter with three tandem ATF sites or from a naturally occurring promoter with ATF4 sites such as E selectin. We also demonstrated a preferential role of the HRG-stimulated mitogen activated protein kinase pathway, but not the phosphatidylinositol 3'-kinase pathway, in supporting the observed increase in ATF4 DNA binding activity and transcription from E-selectin promoter in HRG-stimulated cells. Because ATF4 binding sites are present in a variety of growth-regulating cellular genes, these findings suggest that the stimulation of ATF4 expression and its transactivating functions may constitute an important mechanism of HRG-mediated regulation of putative genes with diversified functions. The present study is the first demonstration of regulation of expression and transactivation ability of ATF4 by any polypeptide growth factor. PMID- 10667577 TI - Protein phosphorylation is a regulatory mechanism for O6-alkylguanine-DNA alkyltransferase in human brain tumor cells. AB - The biochemical regulation of human O6-alkylguanine-DNA alkyltransferase (AGT), which determines the susceptibility of normal tissues to methylating carcinogens and resistance of tumor cells to many alkylating agents, is poorly understood. We investigated the regulation of AGT by protein phosphorylation in a human medulloblastoma cell line. Incubation of cell extracts with [gamma-32P]ATP resulted in Mg(2+)-dependent phosphorylation of the endogenous AGT. Immunoprecipitation after exposure of the cells to 32P-labeled inorganic phosphate showed that AGT exists as a phosphoprotein under physiological conditions. Western analysis and chemical stability studies showed the AGT protein to be phosphorylated at tyrosine, threonine, and serine residues. Purified protein kinase A (PKA), casein kinase II (CK II), and protein kinase C (PKC) phosphorylated the recombinant AGT protein with a stoichiometry of 0.15, 0.28, and 0.44 (mol phosphate incorporated/mol protein), respectively. Residual phosphorylation of the endogenous AGT by the PKs present in cell homogenates and phosphorylation of the recombinant AGT by purified serine/threonine kinases, PKA, PKC, and CK II reduced AGT activity by 30-65%. Conversely, dephosphorylation of cell extracts by alkaline phosphatases stimulated AGT activity. We also identified consensus phosphorylation motifs for many cellular kinases, including PKA and CK II in the AGT protein. These data provide the first and conclusive evidence of AGT phosphorylation and suggest that reversible phosphorylation may control the activity of this therapeutically important DNA repair protein in human normal and cancer cells. PMID- 10667578 TI - Nuclear matrix proteins associated with DNA in situ in hormone-dependent and hormone-independent human breast cancer cell lines. AB - The nuclear matrix is a dynamic RNA-protein complex that organizes chromatin and regulates nuclear DNA metabolism. Nuclear matrix proteins informative in the diagnosis of cancer have been identified. Here, the nuclear matrix breast cancer proteins (NMBCs) cross-linked to nuclear DNA in situ with cisplatin in human breast cancer cell lines were analyzed by two-dimensional gel electrophoresis. We identified NMBCs that were differentially associated with nuclear DNA of hormone dependent and -independent breast cancer cell lines. Three DNA cross-linked NMBCs were found to be exclusive to estrogen receptor-positive, hormone-dependent breast cancer cells, whereas two NMBCs were observed only in estrogen receptor negative, hormone-independent breast cancer cells. Changes in these NMBCs were observed when hormone-dependent breast cancer cells became hormone independent. Furthermore, we show that the intermediate filament protein vimentin is associated with the nuclear DNA of MDA-MB-231 breast cancer cells, an estrogen receptor-negative, hormone-independent breast cancer cell line with high metastatic potential. These nuclear matrix DNA-binding proteins may play important roles in breast tumorigenesis. PMID- 10667579 TI - Chemoprevention of colon cancer by specific cyclooxygenase-2 inhibitor, celecoxib, administered during different stages of carcinogenesis. AB - Epidemiological observations and laboratory research have suggested that nonsteroidal anti-inflammatory drugs (NSAIDs) reduce the risk of colon cancer and that the inhibition of colon carcinogenesis by NSAIDs is mediated through the modulation of prostaglandin production by rate-limiting enzymes known as cyclooxygenases (COXs). Because traditional NSAIDs inhibit both COX-1 and COX-2, these drugs induce side effects, such as gastrointestinal ulceration and renal toxicity, through the inhibition of the constitutive COX-1. Overexpression of COX 2 has been observed in colon tumors; therefore, specific inhibitors of COX-2 could serve as chemopreventive agents. Our previous study has shown that celecoxib, an inhibitor of COX-2, while sparing COX-1, inhibited azoxymethane (AOM)-induced colon tumorigenesis when administered during both initiation and postinitiation stages, ie., celecoxib administered continuously before, during, and after carcinogen treatment. This study examined the dose-response effect of celecoxib when administered during the initiation and postinitiation stages. In addition, the chemopreventive effects of high-dose celecoxib administered during the promotion/progression stage of colon carcinogenesis, ie., continuous celecoxib administration beginning 14 weeks after the carcinogen treatment, was determined in male F344 rats. We also measured the steady-state levels of celecoxib in the plasma of animals given this inhibitor. Groups of 5-week-old male F344 rats were fed either a control diet or experimental diets containing 500, 1000, or 1500 ppm celecoxib. At 7 and 8 weeks of age, rats scheduled for carcinogen treatment were injected s.c. with AOM at a dose rate of 15 mg/kg body weight/week. Groups of animals destined for the promotion/ progression study and initially receiving the control diet were switched to a diet containing 1500 ppm celecoxib beginning 14 weeks after the second AOM treatment. All rats remained on their respective dietary regimens until the termination of the study, ie., 52 weeks, and were then sacrificed. Colon tumors were evaluated histopathologically. Administration of 500, 1000, or 1500 ppm celecoxib during the initiation and postinitiation stages significantly inhibited the incidence (P < 0.01 to P < 0.0001) as well as the multiplicity (P < 0.01 to P < 0.0001) of adenocarcinomas of the colon in a dose-dependent manner. Importantly, administration of 1500 ppm celecoxib during the promotion/progression stage also significantly suppressed the incidence and multiplicity of adenocarcinomas of the colon (P < 0.01). Also, administration of celecoxib to the rats during the initiation and postinitiation periods and throughout the promotion/progression stage strongly suppressed colon tumor volume (P < 0.0002 to P < 0.001). The steady-state plasma concentration of celecoxib increases somewhat with the dose. Thus, in this model system, the chemopreventive efficacy of celecoxib is dose-dependent when this COX-2 inhibitor is administered during the initiation and postinitiation periods. This study provides the first evidence that celecoxib is also very effective when it is given during the promotion/progression stage of colon carcinogenesis, indicating that the chemopreventive efficacy is achieved during the later stages of colon tumor development. This suggests that celecoxib may potentially be an effective chemopreventive agent for the secondary prevention of colon cancer in patients with familial adenomatous polyposis and sporadic polyps. PMID- 10667580 TI - Separating favorable from unfavorable prognostic markers in breast cancer: the role of E-cadherin. AB - Distant metastases are the major cause of morbidity and mortality in women with breast cancer. The ability to predict the metastatic proclivity is essential in choosing the optimal treatment. Tumor size and grade, which are frequently used markers in node-negative breast cancer patients, are inadequate markers for prognosis and individualized treatment design. The steps in metastatic progression include angiogenesis, invasion, and changes in adhesion characteristics. We developed a strategy for choosing biomarkers representing these steps in malignant progression to identify patients with occult metastases who will need chemotherapy and spare those women whose tumors have not developed the capacity to spread. To evaluate the added significance of E-cadherin to that of nm23-H1 and angiogenesis in determining metastatic proclivity, we used archival material from 168 node-negative breast cancer patients who were treated with mastectomy without any adjuvant chemotherapy or hormone therapy. Immunohistochemistry was used to detect E-cadherin and nm23-H1 expression, whereas angiogenesis was determined by microvessel count (MVC) after immunohistochemical staining. The median follow-up is 14 years. We found that E cadherin is better in identifying the poor prognosis patients. The 14-year disease-free survival (DFS) is 84%, 80%, and 56% in patients with high, intermediate, and low E-cadherin. The worst prognosis group using nm23-H1 and MVC as biomarkers has a 14-year DFS of 62%. In this group, if E-cadherin is low, the 14-year DFS is further decreased to 44%. Nm23-H1 and MVC are better in identifying the good prognosis patients. The long-term DFS is >90% if MVC is low or if nm23-H1 is high. Multivariate analysis shows that E-cadherin, nm23-H1, and MVC are more significant prognostic biomarkers than tumor size or grade. Loss of E-cadherin appears to be a latter step in the metastatic progression compared to angiogenesis and the loss of nm23-H1 expression. PMID- 10667581 TI - Association of vitamin D receptor gene polymorphism with prostate cancer and benign prostatic hyperplasia in a Japanese population. AB - Recent studies have suggested that vitamin D is an important determinant of prostate cancer risk and inherited polymorphisms in the 3'-untranslated region (3'UTR) of the vitamin D receptor (VDR) gene are associated with the risk and progression of prostate cancer. This study was conducted to explore the association of VDR gene polymorphisms with prostate cancer risk in Japanese men who are considered to be much less influenced by environmental risk factors for prostate cancer. We studied 222 prostate cancer patients, 209 benign prostatic hyperplasia (BPH) patients, 128 male controls who were over 60 years old and without any evidence of prostate cancer or BPH, and 198 female controls. A PCR RFLP method was used to determine three VDR gene polymorphisms in the 3'UTR characterized by restriction enzymes BsmI, ApaI and TaqI. In the BsmI polymorphism, heterozygosity or homozygosity for the absence of the BsmI restriction site was associated with one-third the risk of prostate cancer (P < 0.0001; odds ratio, 3.31; 95% confidence interval, 2.05-5.32) and with one-half the risk of BPH (P < 0.005; odds ratio, 2.07; 95% confidence interval, 1.33-3.22) compared with the male controls. The TaqI and ApaI polymorphisms did not show any significant association with either prostate cancer or BPH. The results indicate that the BsmI polymorphism in the VDR gene plays a significant role in protection against prostate cancer and BPH. Because of the racial difference in the strength of the linkage disequilibrium between the three polymorphisms, additional studies are required to apply the present results to other racial-ethnic groups. PMID- 10667582 TI - Anti-invasive, antitumorigenic, and antimetastatic activities of the PHSCN sequence in prostate carcinoma. AB - Using naturally serum-free SU-ECM basement membranes as invasion substrates showed that plasma fibronectin was necessary to stimulate invasion by DU 145 human and metastatic MATLyLu (MLL) rat prostate carcinoma cells. This activity mapped to the PHSRN sequence, which induced invasion through alpha5beta1 integrin. PHSCN, a competitive inhibitor, blocked both PHSRN- and serum-induced invasion. Acetylated, amidated PHSCN (Ac-PHSCN-NH2) was 30-fold more potent; however, Ac-HSPNC-NH2 was inactive. Rats receiving injections s.c. with 100,000 MLL cells were treated systemically by i.v. injection three times weekly with 1 mg of either Ac-PHSCN-NH2 or Ac-HSPNC-NH2 beginning 24 h later, three times weekly with 1 mg of Ac-PHSCN-NH2 beginning only after surgery to remove large (2 cm) MLL tumors, or were left untreated. MLL tumors grew rapidly in Ac-HSPNC-NH2 treated and in untreated rats. MLL tumor growth in rats treated with Ac-PHSCN-NH2 beginning 1 day after MLL cell injection was reduced by 99.9% during the first 16 days of treatment, although subsequent tumor growth occurred. MLL tumor cryosections immunostained with anti-PECAM-1 showed that Ac-PHSCN-NH2 inhibited neovascularization by 12-fold during this time. Whether initiated after MLL cell injection or only after MLL tumor removal, Ac-PHSCN-NH2 treatment reduced the numbers of MLL lung colonies and micrometastases by 40- to >100-fold, whereas Ac HSPNC-NH2 was inactive. Thus, Ac-PHSCN-NH2 may be a potent antitumorigenic and antimetastatic agent for postsurgical use prior to extensive metastasis. PMID- 10667583 TI - Radiation-induced apoptosis of endothelial cells in the murine central nervous system: protection by fibroblast growth factor and sphingomyelinase deficiency. AB - Injury to the central nervous system (CNS) by ionizing radiation may be a consequence of damage to the vascular endothelium. Recent studies showed that radiation-induced apoptosis of endothelial cells in vitro and in the lung in vivo is mediated by the lipid second messenger ceramide via activation of acid sphingomyelinase (ASM). This apoptotic response to radiation can be inhibited by basic fibroblast growth factor or by genetic mutation of ASM. In the CNS, single dose radiation has been shown to result in a 15% loss of endothelial cells within 24 h, but whether or not this loss is associated with apoptosis remains unknown. In the present studies, dose- and time-dependent induction of apoptosis was observed in the C57BL/6 mouse CNS. Apoptosis was quantified by terminal deoxynucleotidyl transferase-mediated nick end labeling, and specific endothelial apoptosis was determined by histochemical double labeling with terminal deoxynucleotidyl transferase-mediated nick end labeling and Lycopersicon esculentum lectin. Beginning at 4 h after single-dose radiation, apoptosis was ongoing for 24 h and peaked at 12 h at an incidence of 0.7-1.4% of the total cells in spinal cord sections. Up to 20% of the apoptotic cells were endothelial. This effect was also seen in multiple regions of the brain (medulla, pons, and hippocampus). A significant reduction of radiation-induced apoptosis was observed after i.v. basic fibroblast growth factor treatment (0.45-4.5 microg/mouse). Identical results were noted in C3H/HeJ mice. Furthermore, irradiated ASM knockout mice displayed as much as a 70% reduction in endothelial apoptosis. This study demonstrates that ionizing radiation induces early endothelial cell apoptosis throughout the CNS. These data are consistent with recent evidence linking radiation-induced stress with ceramide and suggest approaches to modify the apoptotic response in control of radiation toxicity in the CNS. PMID- 10667584 TI - Differential toxicities of anticancer agents among DNA repair and checkpoint mutants of Saccharomyces cerevisiae. AB - Most cytotoxic anticancer agents damage DNA directly, interfere with DNA metabolism or chromosome segregation, and are particularly toxic in dividing cells. Although a considerable amount of information on the mechanisms of action of these agents is available, the molecular bases for selective tumor cell killing by chemotherapy are largely unknown. Many genetic alterations found in sporadic and hereditary cancers affect functions in DNA repair and cell cycle control and result in sensitivity to DNA damaging agents. We have therefore set out to determine the effects of these cancer mutations on sensitivity or resistance to various chemotherapeutic agents. Because most of the affected genes are well conserved among eukaryotes, we have carried out a comprehensive analysis of a panel of isogenic yeast strains, each defective in a particular DNA repair or cell cycle checkpoint function, for sensitivity to the Food and Drug Administration-approved cytotoxic anticancer agents. Widely different toxicity profiles were observed for 23 agents and X-rays, indicating that the type of DNA repair and cell cycle checkpoint mutations in individual tumors could strongly influence the outcome of a particular chemotherapeutic regimen. PMID- 10667585 TI - Prostate-specific antigen promoter/enhancer driven gene therapy for prostate cancer: construction and testing of a tissue-specific adenovirus vector. AB - A range of luciferase reporter vectors was constructed, incorporating 5'-flanking sequences from the prostate-specific antigen (PSA), human glandular kallikrein 2 (hKLK2), and cytomegalovirus (CMV) promoters for expression control. Tissue specificity was evaluated in the PSA-positive line LNCaP and PSA-negative cells from different tissues of origin (CoLo320, DG75, EJ, A2780, and Jurkat). The minimal 628-bp PSA and hKLK2 promoters showed only low-level expression in either PSA-positive or PSA-negative cells and showed no increase with the addition of androgen. Tandem duplication of the PSA promoter slightly increased expression in PSA-positive LNCaP cells. The addition of CMV enhancer sequences upstream of a single PSA or hKLK2 promoter substantially but nonspecifically increased luciferase expression in all cell lines tested. However, placing a 1455-bp PSA enhancer sequence upstream of either the PSA or hKLK2 promoters increased expression 20-fold in the PSA-positive cell line LNCaP but not in the PSA negative lines. Tandem duplication of the PSA enhancer increased expression to approximately 50-fold higher than either promoter alone while retaining tissue specific control. The level of expression was reduced by the addition of a third copy of the PSA enhancer. Expression from all enhancer constructs was increased 100-fold above basal levels when induced with the androgen dihydrotestosterone, with the PSA-based constructs consistently exhibiting roughly twice the level of expression of the hKLK2-based constructs at all androgen concentrations. Adenovirus vectors were produced in which either enhanced green fluorescent protein or nitroreductase could be expressed from the optimized PSA double enhancer-promoter construct and evaluated in LNCaP cells and the bladder-derived line EJ. Control vectors with the CMV promoter gave good levels of expression in both cell lines, whereas the PSA constructs only produced detectable levels of protein in the LNCaP cells as assessed by fluorescence of enhanced green fluorescent protein or by Western blotting of nitroreductase. LNCaP but not EJ cells were selectively sensitized to the prodrug CB1954 following infection with Ad-PSA(EEP)-NR. The PSA-based nitroreductase virus produced comparable amounts of nitroreductase and sensitization to CB1954 approaching that of the CMV-driven virus. Plasmid and adenovirus constructs combining PSA enhancer and promoter sequences demonstrate selective expression of linked genes in PSA-positive cells. The expression is induced by androgen and gives therapeutically relevant levels of effector proteins. PMID- 10667586 TI - The geldanamycins are potent inhibitors of the hepatocyte growth factor/scatter factor-met-urokinase plasminogen activator-plasmin proteolytic network. AB - The Met receptor tyrosine kinase and its ligand, hepatocyte growth factor/scatter factor (HGF/SF), have been implicated in human tumor development and metastasis. HGF/SF induces the expression of urokinase plasminogen activator (uPA) and the uPA receptor (uPAR), important mediators of cell invasion and metastasis. We have developed a cell-based assay to screen for inhibitors of this signaling system using the induction of endogenous uPA and uPAR and the subsequent conversion of plasminogen to plasmin as the biological end point. Assay validation was established using a neutralizing antiserum to HGF/SF and a uPA inhibitor (B428), as well as inhibitors of the MKK-MAPK1/2 pathway, shown previously to be important in the induction of uPA and uPAR. Using this assay, we found several classes of molecules that exhibited inhibition of HGF/SF-dependent plasmin activation. However, we discovered that certain members of the geldanamycin family of anisamycin antibiotics are potent inhibitors of HGF/SF-mediated plasmin activation, displaying inhibitory properties at femtomolar concentrations and nine orders of magnitude below their growth inhibitory concentrations. At nanomolar concentrations, the geldanamycins down-regulate Met protein expression, inhibit HGF/SF-mediated cell motility and invasion, and also revert the phenotype of both autocrine HGF/SF-Met transformed cells as well as those transformed by Met proteins with activating mutations. Thus, the geldanamycins may have important therapeutic potential for the treatment of cancers in which Met activity contributes to the invasive/metastatic phenotype. PMID- 10667587 TI - Effective elimination of lung metastases induced by tumor cells treated with hydrostatic pressure and N-acetyl-L-cysteine. AB - In previous studies, we have demonstrated that application of high hydrostatic pressure (P) to tumor cells in the presence of a slow-reacting membrane impermeable cross-linker (CL), 2'-3'-adenosine dialdehyde, can rearrange cell surface proteins into immunogenic clusters. Here, we present evidence indicating that subsequent reduction of surface protein disulfides with N-acetyl-L-cysteine (NAC) further augments the immunogenic potential of PCL-modified tumor cells both in vitro and in vivo. Immunotherapy with PCL+NAC-modified 3LL-D122 Lewis lung carcinoma cells plus i.v. delivery of NAC in mice bearing established lung metastases provoked an antitumor response capable of eradicating the metastatic nodules as demonstrated by restoration of normal lung weight and histology. In addition, immunization with PCL+NAC-modified tumor cells gave rise to a strong delayed-type hypersensitivity recall response against parental D122 cells. We propose that this novel two-prong strategy, based on local immunization with autologous PCL+NAC-modified tumor cells and systemic boosting with NAC, could provide a practical, effective immunotherapeutic regimen for the treatment of human cancer. PMID- 10667588 TI - Ability of systemic interleukin-12 to hamper progressive stages of mammary carcinogenesis in HER2/neu transgenic mice. AB - Previous studies in mice have shown that chronic administration of recombinant interleukin-12 (IL-12) hampers the progression of both chemical- and oncogene dependent carcinogenesis. This suggests that a new preventive strategy may be envisaged for individuals with a genetic risk of cancer or carrying preneoplastic lesions. Starting at progressive stages of mammary carcinogenesis, female BALB/c and FVB mice carrying the activated rat HER2/neu oncogene (BALB-neuT) or the proto-oncogene (FVB-neuN) under the mouse mammary tumor virus promoter received multiple 5-day courses of different doses of IL-12. The times of tumor appearance, multiplicity, and histopathological features of the neoplastic lesions were evaluated. In both BALB-neuT and FVB-neuN mice, 5-day i.p. courses of 50/100 ng of IL-12/day inhibited mammary carcinogenesis when they coincided with the progression of early preneoplastic lesions. Inhibition appears to depend primarily on the ability of IL-12 to interfere with early tumor angiogenesis. Later treatments are much less effective, and daily doses of 10 and 2 ng are useless. The efficacy of early IL-12 courses suggests that they could be used to prevent mammary tumors in individuals at risk, whereas their lower efficacy in later stages of carcinogenesis and the dose range required pose some constraints on their use in the management of overt preneoplastic lesions. Precise understanding of tumor progression means that effective treatments can be commenced relatively late in the life of individuals at risk and that no lifetime administration is required. PMID- 10667589 TI - Use of fluorogenic histocompatibility leukocyte antigen-A*0201/HPV 16 E7 peptide complexes to isolate rare human cytotoxic T-lymphocyte-recognizing endogenous human papillomavirus antigens. AB - Cervical cancer (CaCx) is the second most common female malignancy worldwide and remains a clinical problem despite improvements in early detection and therapy. CaCx and preinvasive cervical intraepithelial neoplasia (CIN3) are strongly associated with infection by human papillomavirus (HPV), particularly types 16 and 18. Two nonstructural viral proteins, E6 and E7, are constitutively expressed in cervical tumors and are crucial for the maintenance of the transformed phenotype. These proteins thus provide attractive targets for immunotherapy of CaCx mediated by CD8+ CTLs. However, reliable detection and generation of HPV specific CTLs in humans has been difficult. Recently, soluble fluorogenic MHC peptide complexes (tetramers) have greatly increased the sensitivity of antiviral and antitumor CTL detection. To examine the feasibility of this approach for detecting HPV-specific CTLs, we constructed a tetramer consisting of HLA-A*0201 and the best studied HPV CTL peptide epitope, HPV 16 E711-20. Between 2 and 12% of short-term HPV 16 E711-2 CTL lines derived from CaCx patients stained highly with the tetramer. Direct ex vivo staining of peripheral blood mononuclear cells revealed CD8+ tetramer+ high cells at low frequencies in both CIN3 patients (1 of 1,260 to 1 of 19,073) and normal controls (1 of 1,855 to 1 of 42,004). However, short-term in vitro stimulation with the HPV 16 E711-20 peptide expanded CD8+tetramer+ cells to a greater extent in the peripheral blood mononuclear cells from CIN3 patients. Furthermore, the tetramer provided a powerful tool to isolate polyclonal and clonal peptide-specific CTLs from an established HPV 16 E7,11-20 specific CTL line. These purified CTLs were able to lyse both peptide-pulsed targets and targets expressing endogenously processed HPV antigens. This tetramer may therefore be useful for selecting rare high-affinity HPV-specific CTLs for the immunotherapy of CaCx. PMID- 10667590 TI - Mutations in the retinoblastoma-related gene RB2/p130 in lung tumors and suppression of tumor growth in vivo by retrovirus-mediated gene transfer. AB - The retinoblastoma (Rb) family consists of the tumor suppressor pRb/p105 and related proteins p107 and pRb2/p130. Recent immunohistochemical studies of the retinoblastoma family of proteins in 235 specimens of lung cancer show the tightest inverse association between the histological grading in the most aggressive tumor types and pRb2/p130. This led us to study a panel of human lung cancers for mutations in the RB2/p130 gene. Mutations in the Rb-related gene RB2/p130 were detected in 11 of 14 (78.5%) primary lung tumors by single-strand conformation polymorphism and sequence analysis. A Moloney leukemia virus-based retroviral system was set up, and a comparable viral concentration of 1 x 10(7) infectious units/ml was obtained. Retrovirus-mediated delivery of wild-type RB2/p130 to the lung tumor cell line H23 potently inhibited tumorigenesis in vitro and in vivo, as shown by the dramatic growth arrest observed in a colony assay and the suppression of anchorage-independent growth potential and tumor formation in nude mice. The tumors transduced with the RB2/p130 retrovirus diminished in size after a single injection, and a 12-fold reduction in tumor growth after RB2/p130 transduction compared with the Pac-transduced tumors (92% reduction, P = 0.003) and lacZ-transduced tumors (93% reduction, P < 0.001) was found to be statistically significant. These findings provide the missing confirmation that RB2/p130 is a "bona fide" tumor suppressor gene and strengthen the hypothesis that it may be a candidate for cancer gene therapy for lung cancer. PMID- 10667591 TI - Genetic alterations disrupting the nuclear localization of the retinoblastoma related gene RB2/p130 in human tumor cell lines and primary tumors. AB - The prototypic tumor suppressor gene, the retinoblastoma gene (RB/ p105), is mutated in a variety of human tumors. However, to date, mutational data on retinoblastoma family members p107 and RB2/p130 in tumors is lacking. We studied the expression of pRb2/p130 by immunocytochemistry and Western blot analysis in a panel of human osteosarcoma and lymphoid cell lines. Only the lymphoid cell lines showed an abnormal cytoplasmic localization of pRb2/p130, suggesting possible alterations within the region of nuclear localization signaling. We screened these cell lines for genetic alterations of the RB2/p130 gene in the region of the putative bipartite nuclear localization signal (NLS). This region is highly homologous with that of the RB/p105 gene. In addition, we screened four primary Burkitt's lymphomas for genetic alterations in the RB2/p130 gene. Naturally occurring mutations, which disrupt the putative bipartite NLS, were found in lymphoma cell lines and primary tumors, but not in the osteosarcoma cell lines, where normal nuclear localization of the protein was detectable. Site-directed mutagenesis and transfection assay using NLS mutants displayed markedly reduced biological activity as measured by flow cytometric analysis. This study clearly describes RB2/ p130 as an important target for mutations and subsequent inactivation in lymphoma pathogenesis, thus validating that RB2/p130 is a classical tumor suppressor gene. PMID- 10667592 TI - Induced micronucleus frequencies in peripheral lymphocytes as a screening test for carriers of a BRCA1 mutation in breast cancer families. AB - Enhanced sensitivity to the chromosome-damaging effects of ionizing radiation is a feature of many cancer-predisposing conditions. It has been suggested that women with breast cancer are deficient in the repair of radiation-induced DNA damage. We have now investigated whether mutagen sensitivity is related to mutations in the breast cancer gene BRCA1. We studied the induction and repair of DNA damage in lymphocytes of women from families with familial breast cancer and breast and ovarian cancer. The mutagens used were gamma-irradiation and hydrogen peroxide and the DNA effects were determined with the micronucleus test and the comet assay. Women with a BRCA1 mutation (n = 12) and relatives without the familial mutation (n = 10) were compared to controls (i.e., healthy women without family history of breast or ovarian cancer; n = 17). Our results indicate a close relationship between the presence of a BRCA1 mutation and sensitivity for the induction of micronuclei. Compared to a concurrent control, 10 of 11 women with a BRCA1 mutation showed elevated radiation sensitivity. Of the 10 related women without the familial mutation, only 2 had clearly enhanced micronucleus frequencies. In addition to the sensitivity toward gamma-irradiation, hypersensitivity toward hydrogen peroxide was also observed, indicating that the mutagen sensitivity is not solely due to a defect in the repair of DNA double strand breaks. In contrast to the results with the micronucleus assay, we found no significant difference between women with and without a BRCA1 mutation with respect to the induction and repair of DNA damage in the comet assay. This finding suggests a normal rate of damage removal and points to a disturbed fidelity of DNA repair as a direct or indirect consequence of a BRCA1 mutation. Our results support the usefulness of induced micronucleus frequencies as a biomarker for cancer predisposition and suggest its application as a screening test for carriers of a BRCA1 mutation in breast cancer families. PMID- 10667593 TI - Atm deficiency causes an increased frequency of intrachromosomal homologous recombination in mice. AB - Ataxia telangiectasia (AT) patients have inactivating mutations in both copies of the ATM gene. The ATM protein that the gene encodes is involved in DNA double strand break (DSB) recognition; in its absence, p53 response to DSBs is delayed and reduced. In addition, AT patients have a high propensity for cancer, and cells from these patients show chromosomal instability. Here, using an in vivo mouse model system with the pink-eyed unstable mutation, we demonstrate that the absence of functional Atm results in a significantly elevated frequency of intrachromosomal recombination resulting in deletion events (wild-type 17.73%, heterozygous Atm 15.72%, and mutant Atm 30.33%). No such increase was observed in mice heterozygous for Atm. These results further advocate the role of ATM in maintaining genomic integrity after the onset of endogenous damage. This system relies on the initiation of events during a relatively short time frame to produce an observable deletion product. AT patients have a lifelong exposure to endogenous damage and perhaps similarly acting external agents. Because 25% of our genome consists of repeated elements, genomic instability due to an increased level of homologous recombination between such repeats, as observed here, may contribute to carcinogenesis in AT patients. PMID- 10667594 TI - A dominant role for the c-Jun NH2-terminal kinase in oncogenic ras-induced morphologic transformation of human lung carcinoma cells. AB - Oncogenic (activated) Ras is a signal transducer that activates multiple effector mediated signaling pathways leading to altered cell morphology, growth and differentiation, and neoplastic transformation. Activating mutations of Ras family genes have been detected in many types of human cancers, including lung cancer. However, the signaling mechanisms by which oncogenic Ras controls cancer cell growth is poorly characterized. This study evaluates the role of two specific signaling pathways, the c-Jun NH2-terminal kinase (JNK) pathway, and the extracellular signal-regulated kinase (ERK) pathway, in oncogenic Ras-induced morphological transformation of NCI-H82 human small cell lung cancer cells. In the NCI-H82 cell line, oncogenic Ras causes a marked and sustained activation of JNK but only has a modest effect on activation of the ERK pathway. The persistent JNK activation is associated with Ras-induced changes in cell morphology and enhanced transforming activity. Furthermore, JNK activation correlates with the induction of c-Jun expression, c-Jun phosphorylation on serines 63 and 73, and increased AP-1 activity. Deregulation of the JNK pathway using a dominant negative mutant of JNK1, JNK1(APF), completely reverses the oncogenic Ras-induced transformed phenotype, including morphological reversion and inhibition of anchorage-independent growth and low-serum growth. Moreover, expression of JNK1(APF) leads to a decrease in c-Jun/AP-1 activity. In contrast, inhibition of ERK activation via a pharmacological approach using a mitogen-activated protein kinase/ERK kinase-specific inhibitor 2-(2'-amino-3'-methoxyphenyl)-oxanaphthalen 4-one is unable to reverse the Ras-induced transformed morphology and c-Jun/AP-1 induction. These results demonstrate that the JNK/c-Jun/AP-1 pathway plays an essential role in mediating oncogenic Ras function in lung carcinoma cells. PMID- 10667595 TI - Inherited predisposition to pancreatic adenocarcinoma: role of family history and germ-line p16, BRCA1, and BRCA2 mutations. AB - Susceptibility to pancreatic adenocarcinoma appears to be linked to germ-line mutations in genes causing various familial cancer syndromes. The objectives of this study were to estimate the proportion of unselected pancreatic cancer patients belonging to hereditary cancer syndrome families and to determine the frequency ofp16, BRCA1, BRCA2, hMSH2, and hMLH1 germ-line mutations in patients with a personal or family history of cancer. The study population consisted of 102 patients with histologically verified pancreatic adenocarcinoma, unselected for age, sex, family history, or ethnic origin. Patients completed a family history questionnaire and provided blood for mutation analysis. Three-generation pedigrees were constructed and classified as high risk/familial, intermediate risk/ familial, intermediate risk/nonfamilial, or low risk according to defined criteria. High- and intermediate-risk cases were analyzed for germ-line mutations using a combination of methods. Thirty-eight of 102 (37%) patients were characterized as high or intermediate risk, and the remainder were classified as low risk. Germ-line mutations were identified in five (13%) of these cases [one in p16 (I49S); one in BRCA1 (5382 insC); and three in BRCA2 (6174delT)]. The BRCA1 and BRCA2 mutations were identified in Ashkenazi Jewish patients. Four of the mutation carriers had strong family histories of the syndromes associated with the mutated genes. No mutations were identified in patients in whom the sole risk factor was a family history of pancreatic cancer, and only one mutation was found among patients with early-onset disease. We conclude that known causes of genetic predisposition are an important risk factor in a small proportion of pancreatic cancer patients, especially if associated with a strong family history of syndromes associated with the disease. The lack of detectable germ-line mutations in most high- and intermediate-risk cases suggests that there are probably additional, as yet unidentified genes predisposing to this disease. PMID- 10667596 TI - Deregulation of the p14ARF/MDM2/p53 pathway is a prerequisite for human astrocytic gliomas with G1-S transition control gene abnormalities. AB - Deregulation of G1-S transition control in cell cycle is one of the important mechanisms in the development of human tumors including astrocytic gliomas. We have previously reported that approximately two-thirds of glioblastomas (GBs) had abnormalities of G1-S transition control either by mutation/homozygous deletion of RB1 or CDKN2A p16INK4A), or amplification of CDK4 (K. Ichimura et al., Oncogene, 13: 1065-1072, 1996). However, abnormalities of G1-S transition control genes may induce p53-dependent apoptosis in cells. Recent investigations suggest that p14ARF is induced in response to abnormal cell cycle entry and results in p53 accumulation by inhibiting MDM2-mediated transactivational silencing and degradation of p53. To investigate the roles of the G1-S transition control system and the p14ARF/MDM2/p53 pathway in the development of astrocytic gliomas, we examined abnormalities of genes involved in these regulatory pathways in a total of 190 primary human astrocytic gliomas of different malignancy grades [136 GBs, 39 anaplastic astrocytomas (AAs) and 15 astrocytomas (As)]. Sixty-seven percent of GBs (91/136) and 21% of AAs (8/39) had abnormalities of the G1-S control system either by mutation/homozygous deletion of RB1, CDKN2A or CDKN2B, or amplification of CDK4. Seventy-six percent of GBs (103 of 136), 72% of AAs (28 of 39), and 67% of As (10 of 15) had deregulated p53 pathway either by mutation of TP53, amplification of MDM2, or homozygous deletion/mutation of p14ARF. When all of the data were combined and compared, 96% of GBs (87 of 91) and 88% of AAs (7 of 8) with abnormal G1-S transition control also had deregulated p53 pathway. Thus, we demonstrate that deregulation of the G1-S transition control system was almost always accompanied by inactivation of the p53 pathway, clearly illustrating the cooperative roles of these two systems in the development/progression of primary human astrocytic gliomas. PMID- 10667597 TI - ZK7, a novel zinc finger gene, is induced by vascular endothelial growth factor and inhibits apoptotic death in hematopoietic cells. AB - Vascular endothelial growth factor (VEGF) inhibits radiation-induced apoptosis in the leukemia cell line, CMK86 (O. Katoh et al., Cancer Res., 55: 5687-5692, 1995). To elucidate the molecular mechanisms underlying this inhibitory effect of VEGF, we attempted to identify a transcription factor involved in the cellular response to VEGF stimulation. We cloned the cDNA of a novel Kruppel-type zinc finger (ZNF) gene, ZK7, from a cDNA library constructed from the human leukemia cell line CMK86 after incubation with VEGF. This cDNA encoded a protein of 289 amino acids, which contained a Kruppel-associated box A-box domain at the NH2 terminus and seven ZNF motifs at the COOH terminus. These ZNF motifs were identical to those of HZF16 (M. Saleh et al., Am. J. Hum. Genet., 52: 192-203, 1993). ZK7 and HZF16 genes appear to be the splice variants transcribed from the same gene. Northern blotting and quantitative reverse transcription-PCR analysis revealed that expression of ZK7 mRNA in CMK86 cells and human hematopoietic progenitor cells was increased after VEGF stimulation, whereas that of HZF16 mRNA remained unchanged. To examine the function of ZK7 protein, we generated clones of a human monocytoid leukemia cell line, U937, which were stably transfected with ZK7 or HZF16 cDNA. Importantly, ZK7-overexpressing cells had lower sensitivity to ionizing radiation and the chemotherapeutic agent etoposide than U937 parent cells or HZF16-overexpressing cells. Therefore, ZK7 protein may be involved in the inhibitory effect of VEGF on apoptotic cell death in human hematopoietic cells. PMID- 10667598 TI - The cancer-free phenotype in trichothiodystrophy is unrelated to its repair defect. AB - The DNA repair-deficient genetic disorders xeroderma pigmentosum (XP) and trichothiodystrophy (TTD) can both result from mutations in the XPD gene, the sites of the mutations differing between the two disorders. The hallmarks of XP are multiple pigmentation changes in the skin and a greatly elevated frequency of skin cancers, characteristics that are not seen in TTD. XP-D and most TTD patients have reduced levels of DNA repair, but some recent reports have suggested that the repair deficiencies in TTD cells are milder than in XP-D cells. We reported recently that inhibition of intracellular adhesion molecule-1 (ICAM-1) expression by UVB irradiation was similar in normal and TTD cells but increased in XP-D cells, suggesting a correlation between ICAM-1 inhibition and cancer proneness. In the first part of the current work, we have extended these studies and found several other examples, including XP-G and Cockayne syndrome cells, in which increased ICAM-1 inhibition correlated with cancer proneness. However, we also discovered that a subset of TTD cells, in which arg112 in the NH2-terminal region of the XPD protein is mutated to histidine, had an ICAM-1 response similar to that of XP-D cells. In the second part of the work, we have shown that TTD cells with this specific NH2-terminal mutation are more sensitive to UV irradiation than other TTDs, most of which are mutated in the COOH-terminal region, and are indistinguishable from XP-D cells in cell killing, incision breaks, and repair of cyclobutane pyrimidine dimers. Because the clinical phenotypes of these patients do not obviously differ from those of TTDs with mutations at other sites, we conclude that the lack of skin abnormalities in TTD is independent of the defective cellular responses to UV. It is likely to result from a transcriptional defect, which prevents the skin abnormalities from being expressed. PMID- 10667599 TI - Interaction with endothelial cells is a prerequisite for branching ductal alveolar morphogenesis and hyperplasia of preneoplastic human breast epithelial cells: regulation by estrogen. AB - Although there is experimental evidence supporting the involvement of angiogenesis in the pathogenesis of breast cancer, the exact nature and effects of interaction between human breast epithelial cells (HBECs) and endothelial cells (ECs) have not been described thus far. This approach requires an assay system that permits growth and differentiation of both epithelial and endothelial cells. Here, we report the development of a three-dimensional in vitro culture system that supports growth and functional differentiation of preneoplastic HBECs and ECs and recapitulates estrogen-induced in vivo effects on angiogenesis and the proliferative potential of MCF10AT xenografts. MCF10A and MCF10AT1-EIII8 (referred to as EIII8) cell lines used in this study are normal or produce preneoplastic lesions, respectively. When MCF10A or EIII8 cells are seeded on reconstituted basement membrane (Matrigel), both lines organize into a three dimensional tubular network of cells; however, tubes produced by EIII8 cells appear multicellular in contrast to unicellular structures formed by MCF10A cells. However, when MCF10A or EIII8 cells are cocultured with human umbilical vein endothelial cells (HUVECs) on Matrigel, rather than interacting with extracellular matrix, the ECs exhibit preferential adherence to epithelial cells. Although both MCF10A and EIII8 cells provide preferential substrate for EC attachment, only EIII8 cells facilitate sustained proliferation of ECs for prolonged periods that are visualized as "endothelial cell enriched spots," which express factor VIII-related antigen. At regions of endothelial-enriched spots, preneoplastic HBECs undergo branching ductal-alveolar morphogenesis that produce mucin, express cytokeratins, and proliferating cell nuclear antigen. The presence of actively proliferating and functional endothelial cells is essential for ductal-alveolar morphogenesis of preneoplastic HBECs because without ECs, the epithelial cells formed only tubular structures. This ability to establish functional ECs and ductal-alveolar morphogenesis is facilitated only by preneoplastic HBECs because normal MCF10A cells fail to elicit similar effects. Thus, a cause-effect relationship that is mutually beneficial exists between EC and preneoplastic HBECs that is critical for generation of functional vascular networks and local proliferative ductal alveolar outgrowths with invasive potential. Both these processes are augmented by estrogen, whereas antiestrogens inhibit these processes. Induction and maintenance of angiogenic phenotype is associated with up-regulation in expression of interleukin 8 and matrix metalloproteinase-2 and estrogen-induced increases in vascular endothelial growth factor and vascular endothelial growth factor receptor 2. This three-dimensional culture model offers a unique opportunity to study endothelial- and epithelial cell-specific factors that are important for ductal-alveolar morphogenesis, angiogenesis, and progression to malignant phenotype. PMID- 10667600 TI - A peptide mimic of E-selectin ligand inhibits sialyl Lewis X-dependent lung colonization of tumor cells. AB - Selectins bind to carbohydrate ligands in a calcium-dependent manner and play critical roles in host defense and possibly in tumor metastasis. To isolate peptides that mimic E-selectin ligands, we screened a phage peptide library using E-selectin as a target molecule. This attempt unexpectedly failed, probably because the binding affinity of E-selectin to its ligand is low. We then took an approach that is analogous to the isolation of anti-idiotype antibodies and were able to isolate peptides that bound to anticarbohydrate antibodies recognizing E selectin ligands. These peptides, enriched for their binding to anti-Lewis A antibody, were found to bind to E-, P- and L-selectins in a calcium-dependent manner. Phage harboring the identified peptide IELLQAR and synthetic peptides having the same sequence inhibited the binding of sialyl Lewis X or sialyl Lewis A oligosaccharides to E-selectin. The adhesion of HL-60 and B16 melanoma cells expressing sialyl Lewis X to E-selectin was also inhibited by the phage displaying IELLQAR peptide. Moreover, i.v. injected IELLQAR peptide inhibited the lung colonization of mouse B16 melanoma and human lung tumor cells expressing sialyl Lewis X. These results demonstrate that it is possible to isolate peptides mimicking carbohydrate ligands by screening the peptides for binding to anticarbohydrate antibodies and then using them to inhibit carbohydrate-dependent experimental tumor metastasis. PMID- 10667601 TI - Thrombospondin-1 promotes alpha3beta1 integrin-mediated adhesion and neurite-like outgrowth and inhibits proliferation of small cell lung carcinoma cells. AB - Although human small cell lung carcinoma (SCLC) cell lines are typically anchorage-independent and do not attach on most extracellular matrix proteins, OH 1, and several other SCLC cell lines attached on substrates coated with thrombospondin-1 (TSP1). SCLC cells grew long-term as adherent cells on a TSP1 coated substrate. Adhesion of SCLC cells on TSP1 was inhibited by heparin, function-blocking antibodies recognizing alpha3 or beta1 integrin subunits, and by soluble alpha3beta1 integrin ligands. SCLC cells extended neurite-like processes on a TSP1 substrate, which was also mediated by alpha3beta1 integrin. Process formation on a TSP1 substrate was specifically stimulated by epidermal growth factor and somatostatin. Adhesion on TSP1 weakly inhibited SCLC cell proliferation, but this inhibition was strongly enhanced in the presence of epidermal growth factor. TSP1 and an alpha3beta1 integrin-binding peptide from TSP1 also inhibited proliferation when added in solution. High-affinity binding of 125I-labeled TSP1 to OH-1 cells was heparin-dependent and may be mediated by sulfated glycolipids, which are the major sulfated glycoconjugates synthesized by these cells. Synthesis or secretion of TSP1 by SCLC cells could not be detected. On the basis of these results, the alpha3beta1 integrin and sulfated glycolipids cooperate to mediate adhesion of SCLC cells on TSP1. Interaction with TSP1 through this integrin inhibits growth and induces neurotypic differentiation, which suggests that this response to TSP1 may be exploited to inhibit the progression of SCLC. PMID- 10667602 TI - A specific sequence of the noncollagenous domain of the alpha3(IV) chain of type IV collagen inhibits expression and activation of matrix metalloproteinases by tumor cells. AB - The invasive properties of melanoma cells correlate with the expression of matrix metalloproteinases (MMPs) and their physiological modulators (tissue inhibitors of metalloproteinase and membrane-type MMPs) and with that of the alphaVbeta3 integrin. We investigated the effect of anterior lens capsule type IV collagen and of the alpha3(IV) collagen chain on the invasive properties of various tumor cell lines (HT-144 melanoma cells, HT-1080 fibrosarcoma cells). We demonstrated that anterior lens capsule type IV collagen or specifically the synthetic peptide alpha3(IV) 185-203 inhibited both the migration of melanoma or fibrosarcoma cells as well as the activation of membrane-bound MMP-2 by decreasing the expressions of MT1-MMP and the beta3 integrin subunit. PMID- 10667604 TI - Processing of cyclin E differs between normal and tumor breast cells. AB - Cyclin E is a G1 cyclin essential for G1 to S-phase transition of the cell cycle with a profound role in oncogenesis. In tumor cells and tissues, cyclin E is overexpressed and present in its lower molecular weight (LMW) isoforms, and it can be used as a prognosticator for poor patient outcome. In this study, we have examined differences in the processing of cyclin E between normal mammary epithelial and breast cancer cell lines. Five NH2-terminally deleted epitope tagged (FLAG) cyclin E vectors were constructed spanning the range of LMW forms observed in tumor cells. These constructs were transfected into normal and tumor cells and analyzed for the production of cyclin E-FLAG protein products by Western blot analysis with FLAG and cyclin E antibodies. Our results show that only tumor cells had the machinery to process these cyclin E-FLAG constructs to their LMW forms, whereas normal cells mainly expressed the full-length unprocessed form of each protein. Tumor and normal cells always process the cyclin E-FLAG protein in the same way as endogenously expressed cyclin E. This phenomenon is consistent with all of the cell lines used, regardless of transfection efficiency, time of processing posttransfection, or method of transfection. Furthermore, measurement of FLAG-associated kinase activity in the transfectants revealed that the protein products of the cyclin E-FLAG constructs are 10 times more active in tumor cells than in normal cells. These studies suggest that the LMW forms of cyclin E detected at a much higher level in tumor cells arise from posttranslational action of a protease. PMID- 10667603 TI - Heregulin regulation of autocrine motility factor expression in human tumor cells. AB - The exposure of cells to growth factors has been shown to induce cytoskeleton reorganization, leading to stimulation of cell motility and invasion. Heregulin beta1 (HRG), a combinatorial ligand for human epidermal growth factor receptor 3 and human epidermal growth factor receptor 4 receptors, is a regulatory secretory polypeptide with a distinctive function in promoting motility and invasiveness of breast cancer cells. In addition to HRG, motility and invasiveness of tumor cells may also involve up-regulation of expression and function of the autocrine motility factor (AMF). Here we explored the possible involvement of AMF in the motility-promoting action of HRG in the MCF-7 breast cancer cell model system. We report that HRG increases the expression of AMF mRNA by 3-8-fold in an actinomycin D-sensitive manner and does not require de novo protein synthesis. The HRG-induced stimulation of AMF expression was inhibited by specific inhibitors of p42/44MAPK and p38MAPK kinases, but not by an inhibitor of the phosphatidylinositol 3'-kinase pathway. Other HRG-responsive human cell lines demonstrated that HRG does indeed significantly up-regulate AMF expression. Furthermore, HRG-stimulated increased motility was partially suppressed by inclusion of an anti-AMF antibody to breast cancer cells, suggesting that a HRG mediated increase in cell motility may be mediated, at least in part, via induction of AMF. The present study is the first demonstration of AMF regulation by a growth factor and suggests a potential role for AMF in HRG regulation of breast cancer cell motility and a novel function of HRG as a regulator of motility factor expression. PMID- 10667606 TI - Correspondence re: J.L. Stanford et al., Polymorphic receptor in the androgen receptor gene: molecular markers of prostate cancer risk. Cancer Res., 57: 1194 1198, 1997. PMID- 10667605 TI - Oncogenes and tumor angiogenesis: differential modes of vascular endothelial growth factor up-regulation in ras-transformed epithelial cells and fibroblasts. AB - A possible link between oncogenes and tumor angiogenesis has been implicated by the finding that expression of various oncogenes, particularly mutant ras, can lead to a marked induction of a potent paracrine stimulator of angiogenesis, vascular endothelial growth factor (VEGF). We sought to determine how oncogenic ras induction of VEGF is mediated at the molecular level and whether the mechanisms involved differ fundamentally between transformed epithelial cells and fibroblasts. Our results suggest that in a subline (called RAS-3) of immortalized nontumorigenic rat intestinal epithelial cells (IEC-18) that acquired a tumorigenic phenotype upon transfection of mutant ras, up-regulation of VEGF occurs in the absence of an autocrine growth factor circuit. The expression of VEGF mRNA and protein by RAS-3 cells was strongly suppressed in the presence of LY294002, an inhibitor of phosphatidylinositol 3'-kinase, but remained largely unaffected in the same cells treated with an inhibitor (PD98059) of mitogen activated protein/extracellular signal-regulated kinase kinase 1 (MKK/MEK-1). This is consistent with the observation that overexpression of a constitutively activated mutant of MEK-1 (AN3/ S222D) in the parental IEC-18 cells did not result in up-regulation of VEGF production. The impact of mutant ras on VEGF expression was also significantly amplified at high cell density, conditions under which RAS-3 cells became less sensitive to LY294002-induced VEGF down regulation. In marked contrast to cells of epithelial origin, ras-transformed murine fibroblasts (3T3RAS) up-regulated VEGF in a manner that was strongly inhibitable by MEK-1 blockade (ie. treatment with PD98059), whereas these cells were relatively unaffected by treatment with the phosphatidylinositol 3'-kinase inhibitor LY294002. In addition, VEGF was up-regulated by 2-3-fold in NIH3T3 cells overexpressing mutant MEK-1. Collectively, the data suggest that the stimulatory effect of mutant ras on VEGF expression is executed in a nonautocrine and cell type-dependent manner and that it can be significantly exacerbated by physiological/ environmental influences such as high cell density. PMID- 10667607 TI - Present and future capabilities of molecular imaging techniques to understand brain function. AB - This article focuses on the use of positron emitting tracers and positron emission tomography (PET) as the most specific and sensitive means for imaging molecular interactions and pathways within the human brain. The concept of the imaging science of PET is developed whereby the key components that contribute to the overall accuracy of the image of molecular activity need to be separately optimized. These include radiolabelling of tracer molecules and ligands with radioisotopes of short radioactive half-life, the search for specific radioligands and tracers, and hence the need to mine molecular databases for molecules suitable for in-vivo imaging. The sensitivity and accuracy of PET scanners need to be advanced along with improvements in the signal-to-noise ratio of the tomographic reconstruction algorithms. Finally, the models used for the analysis of serial time frames of kinetic data need to be developed, the operation of which have to be effected with the minimum of noise propagation. The future use of PET for drug discovery and development is discussed whereby it offers proof principle for assays of in-vivo expression of therapeutic molecular targets as accessed from the blood stream; tissue pharmacokinetics of novel compounds; degree of occupancy of molecular targets; and pharmacodynamic measures of drug action. The future application of PET rests heavily on drug discoverers contributing to discovering specific PET radioligands and tracers in order to provide these assays through in-vivo molecular imaging. PMID- 10667608 TI - Magnetic resonance imaging and magnetic resonance spectroscopy assessment of brain function in experimental animals and man. AB - This paper introduces the basic principles and techniques of functional magnetic resonance imaging (fMRI) and spectroscopy (MRS). Examples are given of single event human fMRI studies on control subjects, and a graded activation protocol applied to Parkinsonian patients. Possibilities are discussed for using fMRI techniques to study the neural substrate of various pharmacological agents, including drugs of abuse. The application of these pharmacological MRI (phMRI) studies to animal models and the associated technical issues are also addressed. The use of MRS in studying brain status and function is reviewed, with particular emphasis on 13C isotopic labelling studies. PMID- 10667609 TI - Imaging studies on the role of dopamine in cocaine reinforcement and addiction in humans. AB - We summarize our studies with positron emission tomography investigating the role of dopamine (DA) in the reinforcing effects of cocaine and methylphenidate in humans and its involvement in cocaine addiction. These studies have shown that the rate at which cocaine and methylphenidate enter the brain and block the dopamine transporters (DAT) is the variable associated with the 'high', rather than the presence per se of the drug in the brain. Our studies also show that, while the level of DAT blockade is important in predicting the intensity of the 'high' induced by these drugs (DAT blockade > 50% is required for these drugs to induce a 'high'), the rate at which DAT are blocked determines whether the 'high' is perceived or not. Thus, oral methylphenidate, which leads to slow DAT blockade, does not induce a 'high', even at doses which block DAT more than 60%. In cocaine abusers, we have shown significant reductions in DA D2 receptors that are associated with decreased metabolism in cingulate gyrus and in orbitofrontal cortex. We suggest that this is one of the mechanisms by which DA disruption leads to compulsive drug administration in cocaine addiction. Cocaine abusers also show significant decreases in DA release, which coupled with the reduction in D2 receptors may result in decreased activation of reward circuits by physiological reinforcers and may perpetuate cocaine use as a means to compensate for this deficit. Thus, strategies to enhance DA brain function in ways that mimic physiological DA activity may be of help in overcoming cocaine addiction. PMID- 10667610 TI - Imaging strategies in depression. AB - Functional brain imaging in depressive disorder aims to map brain areas associated with depressive illness or to define the neurochemical correlates of the illness and its treatment. Regional deficits of functional (neural) activity has been consistently detected in the brains of subjects with ongoing affective symptoms, particularly in prefrontal areas. Understanding how these prefrontal deficits can be related to neuroanatomical circuitry, neurochemistry and normal cognitive functions is the major challenge in taking these findings forward. Although many neurochemical systems remain to be adequately imaged in vivo, significant progress has been made in the development of relevant radiotracers, particularly for select serotonin and dopamine receptor subtypes. For the future, functional brain imaging of the neurochemistry associated with core symptoms and treatment responses will be necessary for an adequate description of the pathophysiology of depressive illness. PMID- 10667611 TI - Radio-imaging in small animals. AB - As the resolution of radio-imaging systems improves, the prospect of in-vivo imaging of small animals becomes more attractive. Purpose-built positron emission tomography (PET) scanners capable of imaging individual tissues within the rat brain are now in routine experimental use and in-vivo tracer and saturation kinetic studies are now possible. The study of small animals in this way does have intrinsic problems and constraints associated with it. For example, the animal must be completely immobilized, stable ligand within the radiolabelled preparation may be limiting and anatomical definition may be poor. In spite of this, consistent, semi-quantitative data can be produced and in-vivo radio imaging can provide a genuine and unique complement to more conventional techniques. Animal numbers can be significantly reduced and the quality of data improved due to reduced inter-animal variation, and longitudinal studies, to monitor disease progression, are feasible. As the resolution of imaging systems improves still further, such studies could be extended to mouse, in addition to rat, models of disease. PMID- 10667612 TI - Dopamine as the wind of the psychotic fire: new evidence from brain imaging studies. AB - Abnormalities of dopamine function in schizophrenia are suggested by the common antidopaminergic properties of antipsychotic medications. However, direct evidence of a hyperdopaminergic state in schizophrenia has been difficult to demonstrate, given the difficulty of measuring dopamine transmission in the living human brain. This situation is rapidly changing. Recent developments in positron emission tomography and single-photon emission tomographic techniques enabled measurement of acute fluctuation of synaptic dopamine in the vicinity of D2 receptors. Using this technique, we, and others, measured the increase in dopamine transmission following acute amphetamine challenge in untreated patients with schizophrenia and matched healthy subjects. Following a brief overview of these new brain imaging techniques, the main results derived with this method in patients with schizophrenia are described: (1) amphetamine-induced dopamine release is elevated in patients with schizophrenia, supporting the idea that schizophrenia is associated with dysregulation of dopamine transmission; (2) following amphetamine, hyperactivity of dopamine transmission is associated with activation of psychotic symptomatology; (3) this dysregulation of dopamine release is not a long-term consequence of previous neuroleptic treatment, and is detected in never-medicated patients experiencing a first episode of the illness; and (4) in contrast, this exaggerated response of the dopamine system to amphetamine exposure is not detected in patients studied during a period of illness stabilization, suggesting that the hyperdopaminergic state associated with schizophrenia fluctuates over time. In conclusion, a hyperdopaminergic state might be present in schizophrenia during the initial episode and subsequent relapses, but not during periods of remission. This finding has important consequences for the development of new treatment strategies for the remission phase. PMID- 10667613 TI - What do brain imaging studies tell us about anxiety disorders? AB - In-vivo neuroimaging allows the investigation of brain circuits involved in the experience of anxiety and of receptor changes associated with anxiety disorders. This review focuses on studies by research groups who have compared brain activation maps in different forms of anxiety and on binding studies of the benzodiazepine-GABA(A) receptor. Activation studies have revealed the involvement of many brain areas depending on the condition and the paradigm. However, the orbitofrontal cortex/anterior insula and the anterior cingulate are implicated in all the studies and may represent the nodal point between somatic and cognitive symptoms of any form of anxiety. Most studies of binding at the benzodiazepine GABA(A) receptor are not interpretable because of substantial methodological problems, however, regional and/or global reductions are the most consistent finding in panic disorder. PMID- 10667614 TI - Comparison of the effects of alpha-methyl-p-tyrosine and a tyrosine-free amino acid load on extracellular noradrenaline in the rat hippocampus in vivo. AB - Peripheral administration of an amino acid load lacking tyrosine and its precursor, phenylalanine, causes a lowering of central tyrosine levels. The aim of the present study was to examine the effects of tyrosine depletion on extracellular noradrenaline using microdialysis. Extracellular noradrenaline was measured in hippocampus of the anaesthetized rat under both baseline conditions (with reuptake inhibitor, desipramine, in the perfusion medium) and following administration of the alpha2-adrenoreceptor antagonist, idazoxan. The tyrosine free amino acid load did not alter either baseline noradrenaline or the twofold rise in noradrenaline evoked by idazoxan compared with saline controls. In contrast, the catecholamine synthesis inhibitor, alpha-methyl-p-tyrosine, caused a marked reduction in baseline extracellular noradrenaline and abolished the rise induced by idazoxan. In conclusion, the present data indicate that under the conditions used, a tyrosine-free amino acid mixture may not be an effective means to interfere with central noradrenaline function. This contrasts with recent findings demonstrating that the tyrosine-depletion approach can be used to decrease presynaptic dopamine function. PMID- 10667615 TI - Effects of CCK-4 infusion on the acoustic eye-blink startle and psychophysiological measures in healthy volunteers. AB - The acoustic startle response (ASR) and a range of psychophysiological parameters were evaluated during a continuous intravenous administration of cholecystokinin tetrapeptide (CCK-4) in healthy volunteers. Subjects (n=28) were randomly assigned to double-blind infusion of either CCK-4 (0.5 mg/60 min) or placebo. The ASR sessions were performed prior to infusion and at 20 min and 50 min after the onset of infusion by recording eye-blink response to a series of acoustic stimuli (110 dB, 40 ms). An effect of CCK-4 on the eye-blink startle was observed in the first half of infusion. CCK-4 produced an increase of eye-blink startle amplitude from baseline values in contrast to the decrease observed at this time point with placebo. A mild increase in anxiety and heart rate followed by fatigue was reported with CCK-4. Administration of CCK-4 produced increases in plasma concentrations of adrenocorticotropic hormone, cortisol, prolactin and growth hormone. The results of this study show that a prolonged intravenous administration of CCK-4 may be a useful challenge method for further studies on the role of CCK system in the modulation of human anxiety and stress response. PMID- 10667616 TI - The effects of the novel anxiolytic drug lesopitron, a full and selective 5-HT1A receptor agonist, on pupil diameter and oral temperature in man: comparison with buspirone. AB - We investigated the effects of two 5-HT1A receptor agonists, buspirone and lesopitron, upon pupil size in human volunteers at an ambient luminance level of 32 Cd m(-2) and in darkness. Pupil diameter was monitored with a binocular infrared television pupillometer, before and after the administration of treatments for 4 h at 20-min intervals. Two experiments were conducted. In Experiment 1, 14 healthy male volunteers participated in seven weekly sessions, each associated with the ingestion of one capsule (buspirone 5, 10 and 20 mg, lesopitron 10, 20 and 40 mg and placebo), according to a double-blind balanced, cross-over design. Both buspirone and lesopitron tended to decrease pupil diameter. In darkness, only the highest dose of buspirone (20 mg) caused a miosis that was statistically significant. However, at the luminance level of 32 Cd m( 2) buspirone 10 and 20 mg evoked statistically significant miotic effects, as did the highest dose of lesopitron (40 mg). The miotic effect was significantly greater at 32 Cd m(-2) than in darkness after each dose of buspirone and the highest dose (40 mg) of lesopitron. In Experiment 2, pupil diameter and oral temperature were monitored with an electronic thermometer at 40-min intervals. Twenty healthy male volunteers participated in two weekly sessions, each associated with the sublingual application of 100 microl hydroalcoholic solution (lesopitron 20 mg, placebo), according to a double-blind balanced cross-over design. Lesopitron caused a significant miosis both in darkness and at the luminance level of 32 Cd m(-2); the miosis was greater at 32 Cd m(-2) than in darkness. Lesopitron tended to decrease oral temperature; this effect however, was not statistically significant. The greater effectiveness on the pupil of lesopitron administered sublingually in a solution indicates the importance of first-pass metabolism in reducing the effectiveness of the drug when administered by the mouth. The miosis observed in both experiments may be due to either a sympatholytic or a parasympathomimetic effect of the drugs, or both. The light dependence of the miosis indicates that the 5-HT1A receptor agonists can modulate the light reflex, possibly via the noradrenergic control of central cholinergic neurones in the Edinger-Westphal nucleus. PMID- 10667617 TI - Effects of d-amphetamine and haloperidol on latent inhibition in healthy male volunteers. AB - Latent inhibition (LI) refers to a retardation of learning about the consequences of a stimulus when that stimulus has been passively presented a number of times without reinforcement. Acute positive-symptom schizophrenics, normal volunteers who score high on questionnaire measures of schizotypy and non-patients or animals treated with dopamine agonists show reduced LI. Neuroleptic drugs, such as haloperidol, administered at low doses, potentiate LI and effectively reverse disruption of LI induced by dopamine agonists in animals. However, a high dose of haloperidol, administered on its own, has been found to reduce LI. We examined the effects on LI of acute oral administration of an indirect dopamine-agonist, d amphetamine (5 mg), and a nonselective dopamine receptor antagonist, haloperidol (5 mg), in normal male volunteers, using an associative learning task. Replicating previous reports, we found that d-amphetamine reduced LI; haloperidol also reduced LI, but only in subjects who scored low on the Psychoticism scale of the Eysenck Personality Questionnaire. In a subsequent study, no effect was found of 2 mg oral haloperidol administration on LI. The effect of 5 mg haloperidol on LI is interpreted as similar to that observed with a high dose of haloperidol in rats. PMID- 10667618 TI - A 12-week study comparing moclobemide and sertraline in the treatment of outpatients with atypical depression. AB - One hundred and ninety-seven outpatients with atypical depression [Atypical Depression Diagnostic Scale (ADDS) score=4] were randomized to 12 weeks of double blind treatment with sertraline or moclobemide in a multicentre, parallel-group clinical trial. Patients were started on either 50 mg/day sertraline or 300 mg/day moclobemide. If the therapeutic response was not satisfactory after 4 weeks, the dose could be increased to either 100 mg/day sertraline or 450 mg/day moclobemide. Primary efficacy evaluations were the 29-item Hamilton Psychiatric Rating Scale for Depression (HAM-D) and the Clinical Global Impression of Improvement (CGI-I) response rate (much or very much improved) at study endpoint. Secondary efficacy evaluations included the ADDS, the Hamilton Anxiety Scale (HAMA), the Leeds Sleep Scale, and the Battelle Quality of Life Battery (BQOLB). In the analysis of the 172 patient efficacy-evaluable population, there was significant baseline to endpoint improvement in all primary and secondary efficacy assessments after treatment with either sertraline or moclobemide. At the endpoint, the proportion of responders on CGI-I, was 77.5% in the sertraline group and 67.5% in the moclobemide group (p=0.052). The baseline to endpoint mean 29-item HAM-D score decreased from 35.9 to 14.5 in the sertraline group and from 36.3 to 16.1 in the moclobemide group. Sertraline also resulted in a significantly (p < 0.05) greater degree of improvement at the endpoint, compared with moclobemide, in the proportion of remitters on the HAMA (total score < or = 7), ADDS Category IID (Rejection Sensitivity), Leeds Sleep Factor 4 (Integrity of Behaviour Following Awakening), and on three dimensions of the BQOLB (Energy/Vitality, Social Interaction and Life Satisfaction). There were no other significant differences between treatment groups. Overall, both medications were well tolerated. In this study, both sertraline and moclobemide improved the symptoms of atypical depression. PMID- 10667619 TI - Evidence-based psychopharmacology 2. Appraising a systematic review: is risperidone better than conventional antipsychotics in the treatment of schizophrenia? AB - Systematic reviews are increasingly used to combine the results of several studies in order to define the treatment effect with a high degree of precision. However, reviews of this nature may lead to spurious results and should not be accepted uncritically. This article, the second in a series on evidence-based psychopharmacology, is intended to illustrate the process of critical appraisal of a systematic review. In this example, a systematic review of studies comparing risperidone with conventional antipsychotics is appraised. Interpretation of the results of the systematic review may be hampered by the way in which studies were selected. PMID- 10667620 TI - Withdrawal syndrome following long-term administration of tamoxifen. PMID- 10667621 TI - Loss of anxiety and increased aggression in a 15-year-old boy taking fluoxetine. PMID- 10667622 TI - Rapid tranquillisation in isolated units, i.m. medication preferable to i.v. PMID- 10667623 TI - Oral support measures used in feeding the preterm infant. AB - BACKGROUND: Evidence that bottle-feeding is a stressor for inefficient preterm infant feeders is seen in untoward changes in the physiologic system and nutritive sucking patterns. OBJECTIVE: To determine whether a therapeutic technique, oral support (cheek and jaw support), would influence the cardiopulmonary functions or nutritive sucking patterns of preterm infants during feeding. METHODS: A crossover repeated measures design was used with 20 preterm infants for a total of 40 bottle-feeding sessions. The Whitney Mercury Strain Gage and a Nonin Cardiopulmonary monitor were used to observe sucking characteristics and cardiopulmonary functions during feeding. RESULTS: Infants not receiving support paused longer (F= 6.37, df= 5, p < .001) and more frequently (F= 5.01, df= 5, p < .001) than supported infants. There were no differences between the groups in the number of sucks and bursts, the burst duration, the stability of the total sucking activity, or the rate of sucking. Oxygen saturation (SaO2) values, heart rate, and respiratory rate did not differ between the groups during feeding. Postfeeding SaO2 levels were lower than prefeeding levels for infants not receiving oral support (t= 0.96, df= 19, p= .03). CONCLUSION: Oral support provided stability for the jaw and fostered the return of the infant's prefeeding SaO2 values, but it did not interfere with cardiopulmonary function during feeding. Further research is needed to determine whether there is a cumulative effect of oral support, and whether it influences state behavior. PMID- 10667624 TI - Influence of attention and judgment on perception of breathlessness in healthy individuals and patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Cognitive processes mediate judgments of sensation intensity and the perception of breathlessness. These processes depend on focused attention to make a determination, which has not been systematically investigated. OBJECTIVE: To examine the effect of attention on the perception of breathlessness given alterations in attentional focus based on the subject's experience with the perception. METHODS: Magnitude estimation techniques with inspiratory airflow resistance were used to examine the influence of variations in attentional focus on the judgments associated with the perception of breathlessness. Two experimental magnitude estimation conditions were used to alter the focus of attention and compare it with traditional techniques as a control condition (CC). The subject's typical breathing pattern (EXP-T) and clearest memory of breathlessness (EXP-M) were used as references in the experimental conditions. RESULTS: Findings revealed a significant main effect for condition (CC vs. EXP-T: F= 4.82, p < .01; CC vs. EXP-M: F= 14.82, p < .01) and an interaction effect for group by condition (CC vs. EXP-T: F = 4.82, p < .03; CC vs. EXP-M: F = 5.15, p < .03). Post hoc analysis revealed significant differences only for the chronic obstructive pulmonary disease (COPD) group in both comparisons. CONCLUSIONS: Findings indicate that different focuses of attention based on previous exposure to sensations have an impact on judgments used to determine the intensity of a sensation given similar presentation of stimuli, and thus contribute to alterations in the perception of breathlessness. PMID- 10667625 TI - Factors associated with participation by Mexican migrant farmworkers in a tuberculosis screening program. AB - BACKGROUND: Tuberculosis is an important public health concern among migrant farmworkers in the United States; providing appropriate screening and treatment is difficult due to their highly mobile existence. PURPOSE: To analyze the relationship between variables (susceptibility, severity, barriers, benefits, cues to action, normative beliefs, subjective norm, attitude, and intention) from the Health Belief Model (HBM) and the Theory of Reasoned Action (TRA) and participation by Mexican migrant farmworkers in a tuberculosis screening program. METHOD: A convenience sample of 206 migrant farmworkers were recruited after a presentation of a tuberculosis education program and were tracked during the administration and reading of the tuberculosis skin test. Participants were interviewed in Spanish by the principal investigator using the Tuberculosis Interview Instrument (TII) developed for this study. RESULTS: Most subjects were male, aged 18-27 years, and had less than a sixth-grade education. Of the 206 subjects, 152 (73.4%) received the skin test, 149 (98%) had the skin test read, and 44 (29.5%) had positive skin tests. Based on logistic regression analysis, the model that best predicted intention included cues to action, subjective norm, susceptibility, and attitude. Participation in screening was best predicted by a model containing only two variables: intention and susceptibility. CONCLUSIONS: In this study, logistic regression analysis revealed that a more parsimonious model than the full HBM and TRA model accurately predicted both intention and behavior. The findings may be helpful in developing tuberculosis education and screening programs for Mexican migrant farmworkers. PMID- 10667626 TI - Gender and short-term recovery from cardiac surgery. AB - BACKGROUND: There has been a lack of agreement regarding whether women have poorer outcomes than men following cardiac surgery. OBJECTIVES: To examine the effect of gender on early recovery from cardiac surgery. METHOD: Using a prospective descriptive design, 60 men and 60 women who had coronary artery bypass and/or valve surgery completed the study by participating in interviews in the immediate preoperative period and monthly through the third postoperative month. Measures of life quality, life satisfaction, expected/perceived recovery, functional status, global health status, and social support were examined. RESULTS: Preoperatively, women were more functionally limited (p = 0.019), and reported lower life satisfaction (p = 0.001) and social support (p = 0.006), than men. At 3 months postoperatively, there were few significant differences in outcome measures though women continued to report lesser social support (p = 0.002); women realized significantly greater improvement than men in functional status (p = 0.008); and neither age nor gender consistently predicted recovery. CONCLUSIONS: Recent studies focusing on gender differences in cardiac surgery recovery indicate fewer differences between men and women than once thought. However, the differences identified in this study (women's significantly greater improvement in functional status, lesser social support, and differences in the nature of work to which women return following their surgery) warrant concern and attention in clinical practice. PMID- 10667627 TI - Leadership styles across hierarchical levels in nursing departments. AB - BACKGROUND: Some researchers have reported on the cascading effect of transformational leadership across hierarchical levels. One study examined this effect in nursing, but it was limited to a single hospital. OBJECTIVES: To examine the cascading effect of leadership styles across hierarchical levels in a sample of nursing departments and to investigate the effect of hierarchical level on the relationships between leadership styles and various work outcomes. METHODS: Based on a sample of eight hospitals, the cascading effect was tested using correlation analysis. The main sources of variation among leadership scores were determined with analyses of variance (ANOVA), and the interaction effect of hierarchical level and leadership styles on criterion variables was tested with moderated regression analysis. RESULTS: No support was found for a cascading effect of leadership across hierarchical levels. Rather, the variation of leadership scores was explained primarily by the organizational context. Transformational leadership had a stronger impact on criterion variables than transactional leadership. Interaction effects between leadership styles and hierarchical level were observed only for perceived unit effectiveness. CONCLUSIONS: The hospital's structure and culture are major determinants of leadership styles. PMID- 10667628 TI - Preventing smoking relapse in postpartum women. AB - BACKGROUND: Although many women quit smoking during pregnancy, the majority resume smoking shortly after giving birth. OBJECTIVES: To test a program to prevent smoking relapse in the postpartum period by comparing the rates of continuous smoking abstinence, daily smoking, and smoking cessation self-efficacy in treatment and control groups. METHODS: In a randomized clinical trial, nurses provided face-to-face, in-hospital counseling sessions at birth, followed by telephone counseling. The target population included women who quit smoking during pregnancy and who gave birth at one of five hospitals. The 254 participating women were interviewed 6 months after delivery and assessed biochemically to determine smoking status. RESULTS: The 6-month continuous smoking abstinence rate was 38% in the treatment group and 27% in the control group (odds ratio [OR] = 1.63, 95% confidence interval [CI] .96 - 2.78). Significantly more control (48%) than treatment (34%) group participants reported smoking daily (OR = 1.80, 95% CI = 1.08 - 2.99). Smoking cessation self-efficacy did not vary significantly between the groups. CONCLUSIONS: Smoking cessation interventions focusing on the prenatal period have failed to achieve long-term abstinence. Interventions can be strengthened if they are extended into the postpartum period. PMID- 10667629 TI - Detecting and explicating interactions in categorical data. PMID- 10667630 TI - Exploring subject-related interactions in repeated measures data using three-mode principal components analysis. PMID- 10667631 TI - Epidemiologic study of pulmonary obstruction in workers occupationally exposed to ethyl and methyl cyanoacrylate. AB - The association between pulmonary obstruction (e.g., asthma) and occupational exposure to methyl cyanoacrylate (MCA) and ethyl cyanoacrylate (ECA) was examined in an occupational cohort of 450 persons at an adhesive production facility in Puerto Rico. Employee medical records containing information on physical examinations and pulmonary function tests (PFTs), as well as occupational histories, on each employee over a period of about 17 yr and industrial hygiene measurements were evaluated. The cohort analysis was based on a Cox proportional hazards model. Workers exposed to ECA or MCA were compared to workers unexposed to these chemicals with respect to their risk of becoming an "incident case." An "incident case" was defined as any person whose PFTs were normal at the time of employment, but later demonstrated an obstructive pattern, which was defined as a decline in the ratio of forced expiratory volume exhaled in 1 s to forced vital capacity (FEV1/FVC) below 70%. A separate case-control analysis was also conducted that compared "suspected cases," defined as all those whose PFTs ever demonstrated an obstructive pattern (e.g., asthma), to persons whose PFTs remained within normal limits throughout their employment with respect to their past peak and cumulative exposures to cyanoacrylates. All of these analyses showed no evidence that exposure to average short-term concentrations of ECA or MCA of less than 0.5 ppm and occasional daily peak exposures of at least 1.5 ppm (usually 10 min or less), with occasional higher concentrations during spills, were associated with an increased risk of pulmonary obstruction. However, the study suggested that persons occupationally exposed to cyanoacrylates were more likely to have some reversible eye or upper airway irritation than persons who were unexposed. PMID- 10667632 TI - Alveolar macrophage cytokine production in response to air particles in vitro: role of endotoxin. AB - The interaction of air particles and alveolar macrophages (AMs) may result in the release of proinflammatory cytokines. Normal mouse AMs were treated with concentrated air particle (CAPs) suspensions in vitro. After 5 h, cytokine release [macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor alpha (TNF-alpha)] and phagocytosis of ambient air particles were measured. CAPs samples collected from urban air (Boston) on different days were used. The CAPs samples and their soluble and solid components caused significant MIP-2 and TNF alpha production. Variability in the potency of samples collected on different days was observed. Trace endotoxin was measured in CAPs samples (EU/mg: 2.3 +/- 0.7, mean +/- SE, n = 10). A majority of biologic activity (cytokine induction) and endotoxin content was associated with the solid components. Neutralization of endotoxin by polymyxin B abrogated >80% of TNF-alpha induction by CAPs samples, but inhibited MIP-2 production by only approximately 40%. The trace endotoxin present in CAPs caused much more MIP-2 production than predicted by concentration alone (28 +/- 8-fold increase, n = 9), indicating synergistic interaction with other AM-activating components of the particles. Data suggest that low levels of endotoxin may interact with air particles to activate lung macrophages. PMID- 10667633 TI - Effect of CYP2E1 induction by ethanol on the immunotoxicity and genotoxicity of extended low-level benzene exposure. AB - Potential additive effects of ethanol consumption, a common life-style factor, and low-level benzene exposure, a ubiquitous environmental pollutant, were investigated. Ethanol is a potent inducer of the cytochrome P-450 2E1 (CYP2E1) enzyme, which bioactivates benzene to metabolites with known genotoxicity and immunotoxicity. A liquid diet containing 4.1% ethanol was used to induce hepatic CYP2E1 activity by 4-fold in female CD-1 mice. Groups of ethanol-treated or pair fed control mice were exposed to benzene or filtered air in inhalation chambers for 7 h/d, 5 d/wk for 6 or 11 wk. The initial experiment focused on immunotoxicity endpoints based on literature reports that ethanol enhances high dose benzene effects on spleen, thymus, and bone marrow cellularity and on peripheral red blood cell (RBC) and white blood cell (WBC) counts. No statistically significant alterations were found in spleen lymphocyte cellularity, subtype profile, or function (mitogen-induced proliferation, cytokine production, or natural killer cell lytic activity) after 6 wk of ethanol diet, 0.44 ppm benzene exposure, or both. This observed absence of immunomodulation by ethanol alone, a potential confounding factor, further validates our previously established murine model of sustained CYP2E1 induction by dietary ethanol. Subsequent experiments involved a 10-fold higher benzene level for a longer time of 11 wk and focused on genotoxic endpoints in known target tissues. Bone marrow and spleen cells were evaluated for DNA-protein cross links, a sensitive transient index of genetic damage, and spleen lymphocytes were monitored for hprt-mutant frequency, a biomarker of cumulative genetic insult. No treatment-associated changes in either genotoxic endpoint were detected in animals exposed to 4.4 ppm benzene for 6 or 11 wk with or without coexposure to ethanol. Thus, our observations suggest an absence of genetic toxicity in CD-1 mice exposed to environmentally relevant levels of benzene with or without CYP2E1 induction. PMID- 10667634 TI - Toxaphene is antiestrogenic in a human breast-cancer cell assay. AB - Toxaphene is a complex mixture of chlorinated bornanes, bornenes, and bornadienes and was a heavily used insecticide in the United States until its use was restricted in 1982. There are conflicting reports regarding the potential for toxaphene to induce estrogenic responses in human and nonhuman animals. Due to the public concern over environmental estrogens, the estrogenicity of toxaphene was examined in a human breast-cancer cell assay, the MCF-7 focus assay, which is based on in vitro postconfluent cell proliferation and tissue restructuring. In this assay, 0.1-1 nM 17beta-estradiol (E2) produces maximum postconfluent proliferation and formation of multicellular nodules or foci. Toxaphene was also tested for its ability (1) to bind the estrogen receptor (ER) in a competitive binding assay using recombinant human ERalpha (rhER) and in a whole-cell competitive ER binding assay, and (2) to alter the catabolism of E2 in MCF-7 cell cultures. Results from the MCF-7 focus assay showed: (1) Toxaphene alone was not estrogenic between the concentrations of 0.5 nM and 10 microM, (2) toxaphene in binary combinations with chlordane, dieldrin, or endosulfan (alpha or beta) was not estrogenic, and (3) toxaphene was weakly antiestrogenic (it reduced the number of foci induced by 0.1 nM and 0.01 nM E2). Results from the competitive binding assays showed that (1) toxaphene alone did not bind rhER or ER in MCF-7 cells, and (2) toxaphene in binary combinations with other pesticides did not bind rhER. Results from the growth assay and radiometric analysis of E2 catabolism showed that (1) toxaphene did not alter the growth rate of MCF-7 cell cultures over 13 d, and (2) toxaphene did not alter the catabolism of E2. In conclusion, results from the MCF-7 focus assay demonstrate that toxaphene is weakly antiestrogenic rather than estrogenic. PMID- 10667635 TI - Edema in chronic venous insufficiency and the effect of modern pharmacotherapy. PMID- 10667636 TI - Implications of the lymphatic system in CVI-associated edema. AB - Lymph drainage is intimately linked with venous drainage in health and disease. Evidence exists to support the view that lymphatics frequently fail in venous disease, particularly in its advanced stages. The implications of this are to increase the risk of infection and the amount of edema, events which can only further the morbidity of venous disease. PMID- 10667637 TI - Edema and leg volume: methods of assessment. AB - Various techniques may be used to assess leg edema. The value of these investigations has been discussed in depth in the consensus statement made in Vaux de Cernay in 1997 and supported by Servier Research Group. These techniques can be classified into three groups: The most simple is leg circumference measurement, which can be assessed by a tape measure, or more rigorously with a Leg-O-Meter. This device is a cheap and reproducible method that has been validated and that takes into account the height at which the circumference has been measured. However, circumference measurement is not always correlated with leg (including foot) volume measurement. The second group of techniques assess leg volume. The most simple method is water displacement volumetry, which has been validated in terms of reproducibility. Several other devices have been used: optoelectronic methods, computed tomography, magnetic resonance imaging (high resolution), dual X-ray absorptiometry. These methods are expensive and not all of them have been validated, but these might be the future investigations of choice. Some other investigations assess immediate variations in volume such as water displacement using dynamic foot volumetry, rheoplethysmography, strain gauge plethysmography, and air plethysmography. The assessment made by these methods (using postural, dynamic, or compressive maneuvers) is more an assessment of the venomuscular pump and/or venous outflow than volume assessment. In conclusion, edema, an early and frequent sign of chronic venous insufficiency (CVI), can be precisely measured by several methods. This measurement can be considered one of the most objective ways of assessing treatment efficacy in CVI associated edema. PMID- 10667639 TI - Pharmacologic aspects of a phlebotropic drug in CVI-associated edema. AB - Several phlebotropic drugs, or edema-protecting drugs, are available, the most important of which are found in the gamma-benzopyrone family (flavonoids). gamma Benzopyrones can be plant extracts, semisynthetic preparations, or synthetic preparations. This family is divided into two different groups: flavones and flavonols, and flavanes (flavanones). The flavone group contains various types of molecule and includes diosmin. Here we discuss the pharmacologic aspects in edema associated with chronic venous insufficiency (CVI) of one of the reference phlebotropic drugs, micronized purified flavonoid fraction (MPFF), a semisynthetic preparation from the diosmin group, which represents the latest improvement in flavonoid formulation. Before we detail the pharmacologic aspects, a brief summary of the pathophysiology of edema in CVI is necessary. Several factors are implicated: the veins, which create the conditions favorable to edema; the microcirculation, which is the site of fluid transfer into the interstitial tissue; and the lymphatics, which have a limited possibility to reduce edema. Major discoveries are currently being made in CVI and the microcirculation. Results of studies show that MPFF decreases capillary permeability and increases capillary resistance, which could partly be explained by inhibition of leukocyte activation, migration, and adhesion. This inhibition is linked to a significant decrease in plasma levels of endothelial adhesion molecules (VCAM-1 and ICAM-1) after MPFF treatment. Thus, the CVI-induced damage to the microcirculation is counteracted by MPFF. The lymphatic system is also improved by MPFF treatment. The lymphagogue activity of MPFF has been demonstrated in experimental animal models and confirmed by microlymphographic measurement in patients suffering from severe CVI. The pharmacologic activity of MPFF in lymphedema was observed in a study using an animal model of acute lymphedema and in a study in patients with upper limb lymphedema secondary to breast cancer treatment. All these findings point to the importance of acting on each factor involved in the formation and maintenance of edema. This pharmacologic activity is indeed reflected by the clinical efficacy on edema observed during treatment with MPFF. PMID- 10667638 TI - Management of venous edema: insights from an international task force. AB - An International Task Force made up of a panel of 16 experts has reviewed and objectively evaluated all aspects of chronic venous disease of the leg (CVDL). All available publications on CVDL from 1983 to 1997 were identified through computerized search in Medline and by a manual search. Next, three different screenings were performed in order to select only relevant papers providing a level of scientific evidence that was considered moderate to strong. Final conclusions and further therapeutic recommendations were made based on these publications. Medication, compression, local therapy, sclerotherapy, and surgery are the accepted available therapeutic options for CVDL. For edema, the following recommendations can be made: edema is an early sign of CVDL, but before starting any treatment, nonvenous causes of edema should be excluded. Medication and compression are the therapeutic options for edema that are accepted by the Task Force. Evaluation of their efficacy is based on objective measures of edema. Several well-conducted, placebo-controlled trials have shown efficacy of drugs such as micronized purified flavonoid fraction, rutosides, calcium dobesilate, and coumarin rutin. Graduated compression stockings have been shown to be effective; compression needs to be exerted at least at 35 mm Hg. Bandages, if properly applied, both fixed and stretched, can produce favorable results. Sclerotherapy or surgery is not indicated unless there is saphenofemoral or saphenopopliteal reflux. In the absence of such reflux or following deep venous thrombosis, there is no evidence to support sclerotherapy or surgery. PMID- 10667640 TI - Clinical efficacy of micronized purified flavonoid fraction (MPFF) in edema. AB - Swelling is one of the most frequent complaints of patients in daily clinical practice; leg edema is its objective confirmation. It can be associated with several diseases. Micronized purified flavonoid fraction (MPFF) is a phlebotropic drug commonly used to treat the signs and symptoms associated with chronic venous insufficiency (CVI). It has confirmed its clinical efficacy in different groups of patients suffering from edema: idiopathic cyclic edema, CVI-associated edema, postmastectomy lymphedema and might be beneficial in some of drug-induced edema. In a double-blind, placebo-controlled, randomized study including 30 outpatients suffering from idiopathic cyclic edema syndrome, MPFF 1000 mg/day for 6 weeks, normalized the capillary permeability, assessed by the Landis isotope test, in 3 out of 4 patients (p=0.01). The decrease in capillary hyperpermeability led to a clinically significant decrease in weight and edema. In 200 patients with functional or organic venous insufficiency of the lower limbs, a double-blind, placebo-controlled, randomized study with MPFF 1000 mg/day for 2 months provided strong evidence of a marked improvement in symptoms and signs. A significant reduction in supramalleolar edema (assessed by circumference measurement) was observed, whatever the origin of CVI: functional or organic. MPFF efficacy was also demonstrated in another randomized, multicenter controlled trial in 320 patients suffering from chronic venous insufficiency. In this study, a significant decrease in circumference of the most affected leg was observed after 2 months of treatment (p<0.001), whatever the schedule of administration of MPFF (1000 mg once daily or bid). The benefit of MPFF on edema has been further confirmed by the volometer technique (opto-electronic measuring system) which was performed in a population of 30 patients suffering from CVI and treated by MPFF 1000 mg/day over a 6-week period. The mean volume of the more affected lower leg decreased significantly after a 6-week period of treatment, correlating to a significant improvement in clinical symptoms. MPFF has been also tested on another type of edema, upper limb lymphedema secondary to mastectomy, during a double-blind, placebo-controlled, randomized study in 104 patients. MPFF 1000 mg/day improved all lymphoscintigraphic parameters such as half-life and clearance of the labelled colloid. With regard to evolution of lymphedema volume, a tendency in favor of MPFF was observed in the subgroup of patients with more severe lymphedema. Based on its action on capillary hyperpermeability, MPFF has been used with attractive results when combined with classic treatment for a pilot study carried out in patients with advanced breast cancer (n=21) or ovarian carcinoma (n=3), treated with docetaxel, which causes severe edema as a side effect, even when associated with corticoids. Further trials are under way to assess the possible benefit of MPFF in such patients. These results in different types of edema confirm that, by acting on all parameters involved in edema, veins, lymphatics, and microcirculation, MPFF represents a drug of choice for treating CVI-associated edema. PMID- 10667641 TI - RELIEF study: first consolidated European data. Reflux assEssment and quaLity of lIfe improvement with micronized Flavonoids. AB - The RELIEF study (Reflux assEssment and quality of life improvement with micronized Flavonoids in chronic venous insufficiency [CVI]) is a prospective, controlled, multicenter, international study performed in patients with or without venous reflux. This study was conducted between March 1997 and December 1998 in 23 countries worldwide with the participation of more than 10,000 patients suffering from CVI. The European countries, the subject of this report, were represented by the Czech and Slovak Republics, Hungary, Poland, Russia, and Spain. The principal aims of the study were: 1. To validate the first quality-of life scale specific to chronic venous insufficiency (CMVIQ) in different languages and to assess the evolution of quality of life in patients suffering from CVI, with or without venous reflux, treated with micronized purified flavonoid fraction (MPFF*) (1,000 mg/day). 2. To collect international epidemiologic data on venous reflux assessed with pocket Doppler and photoplethysmography. 3. To assess the evolution of symptoms and signs with a specific emphasis on edema through validated Leg-O-Meter measurement (heaviness, pain, cramps, sensation of swelling, edema) in patients suffering from CVI and treated with MPFF, 1,000 mg/day, during 6 months. The first country-by-country statistical analysis and the European consolidated analysis are now available. The CIVIQ questionnaires adapted to each participating country have been validated with highly significant validity and reproducibility (p<0.0001). All dimensions have demonstrated a highly significant and evolving improvement during the study. The results show several interesting findings concerning the epidemiologic data and, of these, two were particularly interesting: - More than 50% of patients suffering from CVI (class 0 to 4 of the CEAP classification) were reflux-free, which means that they were suffering from functional CVI. Patient distribution between the different classes of the CEAP classification changed in a statistically significant manner after 6 months' treatment with MPFF; the number of patients in the more severe classes decreased to the benefit of the less severe classes. Symptoms such as pain, leg heaviness, sensation of swelling, and cramps were significantly improved (p=0.0001). This was associated with a significant decrease in edema, when present, measured by leg circumferences with the Leg-O-Meter (p=0.0001). In conclusion, the European results of the RELIEF study showed the perfect validity and reproducibility of CIVIQ questionnaire adaptations, and the positive progression of quality-of-life scores on MPFF treatment. This progression was paralleled by clinical improvement of patients reflected not only by assessment of CVI symptoms and signs but also by evolution of the CEAP classification. PMID- 10667642 TI - The relative influence of lesion length and other stenosis morphologies on procedural success of coronary intervention. AB - As coronary interventional technology improves, the influence of lesion length (LL) on procedural success and device selection may vary. Thus, the authors prospectively analyzed 957 consecutive coronary interventions (CI) in 1,404 stenoses to ascertain the influence of lesion length on CI outcome. Stenosis morphology was prospectively classified by the AHA/ACC criteria. LL was analyzed both as dichotomous (S: < 10 mm, L: > 10 mm) variables and by the three-tiered AHA/ACC criteria (I: < 10 mm, II: 10-20 mm, III: > 20 mm). There was a significant univariate relationship between CI success and S stenosis (S: 95.8% vs L: 91.8%, p = 0.002 and I: 96.0%, II: 91.7%, III: 89.3%). Numerous interrelationships involving the morphologic characteristics were noted: lesion morphologies associated with S lesions were concentric (p = 0.0001) and had smooth contour (p = 0.0001), ostial location (p = 0.05) and little calcification (p = 0.0007), while irregular contour (p=0.0001), calcification (p=0.0076), eccentric (p=0.0001), thrombus (p = 0.0001), recent (p = 0.0001) or chronic (p = 0.001) total occlusion were associated with L lesions. When these relationships were taken into account by multiple logistic regression analysis, lesion length was not predictive of procedural outcome (p = 0.099). One morphologic type was associated with increased CI success: irregular contour (p = 0.022); recent (p < 0.0001) or chronic (< 0.0001) occlusions were associated with decreased CI success. Another factor considered was device selection: S lesions were associated with greater balloon angioplasty usage (p = 0.002), whereas more coronary stents (p = 0.024) and rotoblator (p = 0.018) devices were used in L lesions. More balloon angioplasty was performed in concentric (p < 0.0001) lesions; interventional devices were employed more often in eccentric (p < 0.0001) and irregular lesions (p < 0.0001). More complications were noted in lesions with thrombus (p = 0.0002), but lesion length was not predictive (p = NS). Lesion length is not a significant predictor of procedural success when adjusted for other lesion morphologies in the modern interventional era. The availability of new devices has improved the results in longer lesions since the AHA/ACC criteria were originally proposed. PMID- 10667643 TI - Risk factors of atherosclerosis and aortic pulse wave velocity. AB - Aortic pulse wave velocity (PWV) is a noninvasive technique that can estimate aortic stiffness or organic change quantitatively. The authors examined the correlation between age and the PWV value in 113 subjects and also examined the relationship between atherosclerotic associated diseases and PWV. A positive correlation was observed between age and the PWV value. No significant difference was found in the PWV value between groups with and without risk factors of atherosclerosis. No significant difference was observed in the PWV value between groups with and without a history of atherosclerotic disease. PMID- 10667645 TI - Enalapril and losartan augment endogenous nitric oxide release in Takayasu's arteritis--a case report. AB - Prognosis in Takayasu's arteritis is limited owing to renovascular hypertension. The authors report a patient with Takayasu's arteritis who had been unilaterally nephrectomized and presented with malignant hypertension due to renal artery stenosis. Hypertension was refractory to conventional antihypertensive treatment, and stenosis was not accessible by interventional angioplasty. Initiation of enalapril and losartan therapy was successful in improving blood pressure without deterioration of renal function due to ischemic failure. Antihypertensive treatment resulted in dramatically stimulated endogenous nitric oxide (NO) synthesis, while elevated plasma endothelin-1 levels were unchanged. Renovascular hypertension in Takayasu's arteritis is associated with an imbalance of vasoconstrictor peptide endothelin-1 and vasodilator peptide NO. Successful treatment of hypertension by enalapril or losartan results in improved endogenous NO synthesis, which putatively counterbalances excessive vasoconstrictor actions and may retard the progression of renal failure. PMID- 10667644 TI - Clinical assessment of vascular thrombosis using indium-111 platelet scintigraphy. AB - The authors reviewed the clinical value of platelet scintigraphy by using autologous platelets labeled with indium-111-oxine to detect thrombotic activity at vascular thrombosis sites. Thirty-nine patients with deep vein thrombosis, 28 with arteriosclerosis obliterans, and 10 with pulmonary embolism were the subjects of this study. Platelet accumulation on scintigrams had a tendency to correlate with aggravation of acute thrombotic symptoms in deep vein thrombosis and arteriosclerosis obliterans. In addition, appropriate thrombolytic and anticoagulant therapy resulted in reduced platelet accumulation in conjunction with improvement of acute symptoms. On the other hand, this type of scintigraphy was not so sensitive for assessment of pulmonary embolism. Platelet scintigraphy could facilitate the assessment of thrombotic activity and might be useful for determining the optimal indication of thrombolytic and anticoagulant therapy for acute vascular thrombosis. PMID- 10667646 TI - Lamellar bone formation in an atherosclerotic plaque of the carotid artery, with a review of histogenesis--a case report. AB - Longer existing atherosclerotic lesions may contain calcifications; lamellar bone rarely develops within them. A 59-year-old man was referred with a progressive stroke. A high-grade stenosis of the left common carotid artery, formed by an ulcerating atherosclerotic plaque with a free-floating thrombus, was detected on angiography. An urgent endarterectomy was performed. Surprisingly this plaque contained pieces of lamellar bone, proved by histologic examination. PMID- 10667647 TI - Successful percutaneous balloon mitral valvuloplasty using left ventricular pressure as a guide to cross the mitral valve--a case report. AB - Percutaneous balloon mitral valvuloplasty (PBMV) provides an effective alternative to surgery in a selective group of patients with symptomatic mitral stenosis. The Inoue balloon technique involves transseptal catheterization followed by catheter manipulation to cross the mitral valve. The authors describe a case of successful percutaneous balloon mitral valvuloplasty in a patient with severe mitral stenosis and pulmonary hypertension. Left ventricular systolic pressure was used as a guide to locate and to advance the balloon catheter across the mitral valve. This technique to cross the mitral valve has not been reported in the literature. PMID- 10667648 TI - The Fleming Unified Theory of Vascular Disease: a link between atherosclerosis, inflammation, and bacterially aggravated atherosclerosis (BAA) PMID- 10667649 TI - Acute generalized urticaria leading to acute myocardial infarction. PMID- 10667650 TI - Ultra high resolution imaging of the human head at 8 tesla: 2K x 2K for Y2K. AB - PURPOSE: To acquire ultra high resolution MRI images of the human brain at 8 Tesla within a clinically acceptable time frame. METHOD: Gradient echo images were acquired from the human head of normal subjects using a transverse electromagnetic resonator operating in quadrature and tuned to 340 MHz. In each study, a group of six images was obtained containing a total of 208 MB of unprocessed information. Typical acquisition parameters were as follows: matrix = 2,000 x 2,000, field of view = 20 cm, slice thickness = 2 mm, number of excitations (NEX) = 1, flip angle = 45 degrees, TR = 750 ms, TE = 17 ms, receiver bandwidth = 69.4 kHz. This resulted in a total scan time of 23 minutes, an in plane resolution of 100 microm, and a pixel volume of 0.02 mm3. RESULTS: The ultra high resolution images acquired in this study represent more than a 50-fold increase in in-plane resolution relative to conventional 256 x 256 images obtained with a 20 cm field of view and a 5 mm slice thickness. Nonetheless, the ultra high resolution images could be acquired both with adequate image quality and signal to noise. They revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter. DISCUSSION: The elevated signal-to-noise ratio observed in ultra high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the histological level under in vivo conditions. However, brain motion is likely to degrade the useful resolution. This situation may be remedied in part with cardiac gating. Nonetheless, these images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods. PMID- 10667651 TI - Thoracic involvement of systemic lupus erythematosus: clinical, pathologic, and radiologic findings. AB - Thoracic involvement occurs more frequently in systemic lupus erythematosus than in any other connective tissue diseases, and more than half of patients with the disease suffer from the involvement. Primary intrathoracic manifestations include pleural disease (effusions and/or thickening), acute lupus pneumonitis, subacute interstitial lung disease including bronchiolitis obliterans organizing pneumonia and non-specific interstitial pneumonia with fibrosis, chronic interstitial lung disease of usual interstitial pneumonia, pulmonary hemorrhage, pulmonary vascular disease, small airway disease of bronchiolitis obliterans, and pulmonary arterial hypertension. Secondary intrathoracic manifestations include atelectasis due to diaphragmatic dysfunction, opportunistic pneumonia, drug and oxygen toxicity, aspiration, and pleuropulmonary consequences of cardiac and renal failure. PMID- 10667652 TI - CT appearance of implanted esophageal stents. AB - Three different types of esophageal stents, the Z-stent, Ultraflex, and Wall stent, exhibit different shapes on CT, which may suggest a difference in the radial forces applied by each of the stents. CT is useful for displaying the relationship between an esophageal stent and adjacent structures and complications. PMID- 10667653 TI - Pulmonary fat embolism syndrome: CT findings in six patients. AB - PURPOSE: Our purpose is to describe the CT findings in pulmonary fat embolism syndrome (FES). METHOD: Chest radiographs and CT scans of six patients with pulmonary FES were reviewed. Initial and follow-up CT findings were noted, and the extent of CT abnormalities was correlated with partial pressure of arterial oxygen (PaO2). RESULTS: Focal areas of consolidation or ground-glass opacity and nodules were seen in all patients, predominantly in the upper lobes of the lungs. Association between these opacities and pulmonary vessels was indicated in three patients. In the lower lobes of all patients, gravity-dependent opacities predominated. Diffuse ground-glass opacity was noted in five patients. Follow-up CT scans showed rapid improvement in three patients, but the gravity-dependent opacity progressed. The extent of CT abnormalities correlated positively with PaO2 (r = 0.8, p < 0.05). CONCLUSION: CT findings reflect the pathophysiology of this syndrome, which differs from that of simple capillary permeability pulmonary edema. PMID- 10667654 TI - B-cell lymphoma of bronchus-associated lymphoid tissue (BALT): CT features in 10 patients. AB - PURPOSE: The purpose of this work was to describe the CT findings of pathologically confirmed bronchus-associated lymphoid tissue (BALT) lymphoma in 10 patients. METHOD: The CT examinations of 10 patients with pathologically proven BALT lymphoma were reviewed retrospectively by two radiologists. Evaluated findings included number and distribution of lesions. We also assessed other CT findings such as presence of airspace consolidation, nodules, ground-glass attenuation, bubble-like radiolucencies, air bronchogram, bronchial dilatation, and lymphadenopathy. RESULTS: Pulmonary lesions were revealed as airspace consolidation in six patients (60%) and nodule(s) in six (60%). Multiplicity of disease was seen in seven patients (70%) and bilateral lung lesions in six (60%). Areas of ground-glass attenuation were seen in seven patients (70%). Bubble-like radiolucencies were present in five patients (50%) and air bronchogram in nine (90%). Findings of bronchial dilatation and lymphadenopathy were seen in three patients (30%). CONCLUSION: BALT lymphomas usually appear as airspace consolidation or nodules with air bronchogram or adjacent ground-glass attenuation at CT. These findings are similar to previous descriptions of pseudolymphomas. Multiple bilateral lesions are common in BALT lymphoma. Bubble like radiolucencies have not been described previously and can be an additional finding of BALT lymphoma. PMID- 10667655 TI - Primary tumors and mediastinal lymph nodes after neoadjuvant concurrent chemoradiotherapy of lung cancer: serial CT findings with pathologic correlation. AB - PURPOSE: The purpose of this work was to describe the changes of primary tumor and mediastinal lymph nodes on CT after neoadjuvant concurrent chemoradiotherapy and to correlate the CT findings with pathology. METHOD: Twenty-one consecutive patients [N2 disease (n = 19) or resectable T4 and N2 disease (n = 2)] with non small cell lung cancer underwent neoadjuvant concurrent chemoradiotherapy. Changes of primary tumor and mediastinal nodes before and after the therapy were assessed using CT. The CT findings were correlated with pathologic findings. RESULTS: With neoadjuvant therapy, decrease in T stage was achieved in 9 of 21 (43%) patients on CT. On pathology, the remaining tumor consisted mostly of fibrosis and necrosis with little proportion of viable tumor cells (mean volume 17%, range 0-55%). Decrease in nodal stage was achieved in 14 of 21 (67%) patients on pathologic examination. Seven patients had cancer cells in mediastinal lymph nodes: in 6 of 9 (67%) patients with adenocarcinoma and 1 of 12 (8%) patients with squamous cell carcinoma (p = 0.016). CONCLUSION: With neoadjuvant concurrent chemoradiotherapy, the remaining tumor consists mostly of fibrosis or necrosis. Decreased nodal stage on pathology is achieved especially in patients with N2 disease of squamous cell carcinoma. The CT findings of the tumor and mediastinal nodes are not helpful in predicting the pathology after the therapy. PMID- 10667656 TI - Serial high resolution CT findings in nonspecific interstitial pneumonia/fibrosis. AB - PURPOSE: The purpose of this work was to evaluate the radiographic and serial high resolution CT (HRCT) findings in patients with nonspecific interstitial pneumonia/ fibrosis (NSIP). METHOD: We identified 15 patients with biopsy-proven NSIP. Radiography and initial and follow-up CT findings were reviewed. RESULTS: Predominant radiographic findings were bilateral infiltrates distributing in the middle and lower lung zones and decreased lung volumes. At initial CT, predominant patterns were peribronchovascular interstitial thickening (n = 6), parenchymal bands (n = 8), intralobular interstitial thickening (n = 12), and traction bronchiectasis (n = 14). Mixed pattern of ground-glass opacity and consolidation (n = 11) were predominant findings of increased lung opacity. At follow-up CT in 14 cases, the abnormalities had disappeared completely in 3, improved in 9, persisted in 1, and worsened in 1. CONCLUSION: The pulmonary abnormalities observed in NSIP on HRCT can disappear or be diminished in most cases after corticosteroid therapy. Intralobular interstitial thickening and traction bronchiectasis, which have been considered to be indicators of irreversible fibrosis, also show favorable responses. PMID- 10667657 TI - Blood clots mimicking peripheral intrabronchial tumors in patients with hemoptysis: CT and bronchoscopic findings. AB - We report radiographic and clinical findings in two cigarette-smoking patients presenting with hemoptysis. On CT, both patients had peripheral intrabronchial masses together with parenchymal opacities. Bronchoscopy revealed the intrabronchial masses to be blood clots and the parenchymal opacities to correspond to areas of parenchymal hemorrhage. Our cases are novel in that both bronchial and parenchymal sequelae of hemorrhage were simultaneously visualized by CT. Also, our findings suggest that bronchial blood clots should be included in the differential diagnosis of peripheral intrabronchial lesions, notably in patients presenting with hemoptysis. PMID- 10667658 TI - Carbamate poisoning: high resolution CT and pathologic findings. AB - Carbamate insecticides are commonly used agricultural insecticides. The major cause of morbidity and mortality in acute carbamate poisoning is respiratory failure associated with pulmonary edema. Although carbamate poisoning is well recognized in the clinical literature, the findings of high resolution CT (HRCT) with carbamate have not been reported. We report the radiographic and HRCT findings of a patient with acute carbamate poisoning who had pathologically proven interstitial pneumonitis after resolution of initial pulmonary edema. PMID- 10667659 TI - High FDG uptake in a schwannoma: a PET study. PMID- 10667660 TI - "Bridging vascular sign" in the MR diagnosis of exophytic uterine leiomyoma. AB - PURPOSE: The purpose of this study was to evaluate the usefulness of the "bridging vascular sign" in the diagnosis of an exophytic uterine leiomyoma and differentiation of a uterine leiomyoma from some other mass arising in the adnexa on pelvic MRI. METHOD: Of 52 women with a pelvic mass in whom pelvic MRI was performed to determine its origin, 26 women with surgicopathologically proven leiomyomas were included in this study. The other 26 women were proved to have indeterminate adnexal masses such as tuboovarian abscess, endometrioma, fibroma, etc. To obtain axial/ sagittal T2-weighted and pre-/postcontrast T1-weighted images, a 1.5 T unit was used. Positive bridging vascular sign was defined as the presence of curvilinear tortuous signal void vascular structures crossing and/or between the uterus and the pelvic mass. For the presence or absence of this sign, the percentages of patients in each group were calculated and compared. RESULTS: Bridging vascular sign was present in 20 (76.9%) of 26 cases of leiomyomas but in no cases of other adnexal masses. Only exophytic uterine leiomyomas had this sign. CONCLUSION: Bridging vascular sign on MRI may be a useful radiologic sign in the diagnosis and differentiation of an exophytic uterine leiomyoma from some other mass arising in the adnexa. PMID- 10667661 TI - Focal nodular hyperplasia: natural course observed with CT and MRI. AB - PURPOSE: The purpose of this work was to assess the natural course of biopsy proven focal nodular hyperplasia (FNH). METHOD: Eighteen biopsy-proven FNHs in 14 patients (12 women and 2 men) who were followed for at least 6 months with CT and/or MRI were included in the study. The volume of the lesions was calculated twice by two observers using the summation of areas method. Intra- and interobserver variability was assessed by intraclass correlation coefficients. Longitudinal data analysis was performed with generalized estimating equations. RESULTS: The volume of FNH was stable in 6 cases, decreased in 10 cases, and increased in 2 cases. Intra- and interobserver variability in size measurements was 5-10%. Intraclass correlation coefficients were >0.992. Longitudinal data analysis showed that there was a general trend of lesion regression. CONCLUSION: Long-term follow-up and objective measurements performed in patients with biopsy proven lesions show that the natural course of FNH is variable. In particular, lesion regression is not rare. PMID- 10667662 TI - Normal enhancement of the small bowel: evaluation with spiral CT. AB - PURPOSE: The purpose of this work was to determine normal contrast enhancement of the small bowel with biphasic spiral CT, using water as oral contrast agent. METHOD: Biphasic spiral CT was performed in 50 healthy patients undergoing evaluation as potential renal donors. All patients received 500 ml of water as oral contrast agent and 150 ml of Omnipaque 350 administered by mechanical injector at a rate of 3 ml/s. Dual phase CT of the abdomen was performed in each patient. Acquisition of early phase images began 30 s after the start of the intravenous injection, and portal phase images were obtained 60 s after initiation of the contrast agent injection. Attenuation measurements (in Hounsfield units) were obtained from the wall of the small bowel (duodenum, jejunum, ileum) in both the arterial and the portal phases. RESULTS: During the arterial phase, the mean (95% confidence interval) attenuation of the duodenum, jejunum, and ileum was 120 (+/- 5), 119 (+/- 5), and 118 (+/- 5) HU, respectively. During the portal phase, the average attenuation of the duodenum, jejunum, and ileum was 111 (+/- 4), 111 (+/- 3), and 107 (+/- 3) HU, respectively. There was no statistically significant difference between the attenuation of the duodenum, jejunum, or ileum within either the arterial or the portal venous phases. There was a statistically significant difference in small bowel enhancement between the arterial and portal venous phases. CONCLUSION: There is no important variation in small bowel attenuation during the 30 and 60 s scanning phases. This study serves as a normal reference that may be helpful when spiral CT is used to evaluate ischemic bowel or inflammatory small bowel diseases. PMID- 10667663 TI - Brenner tumor of the ovary: CT and MR findings. AB - PURPOSE: The purpose of this study was to determine the CT and MR characteristics of Brenner tumors, rare epithelial neoplasms of the ovary. METHOD: CT and MR scans of eight pathologically proven Brenner tumors of the ovary (seven benign, one malignant, and one associated with mucinous cystadenoma) were retrospectively reviewed. The masses were analyzed for location, size, external configuration, internal architecture, enhancement pattern, presence of calcification, and metastatic spread. RESULTS: The patients' median age was 63 years (range 39-79 years), and the mean size of the tumors was 11.4 cm (7.5-17 cm). All tumors were unilateral and had a well-defined margin. The mass was mostly solid in three, mostly cystic in one, and "mixed" solid and cystic in four cases. The tumors with cystic components (n = 5) were mostly multilocular in appearance (n = 4). All the solid components showed mild homogeneous enhancement on postcontrast CT and MRI. Extensive amorphous calcification within the solid component on CT was seen in five of six cases (83%). No characteristic findings discriminating malignancy from benign Brenner tumor could be found. CONCLUSION: Extensive amorphous calcification in a solid mass or solid component in a multilocular cystic mass is a characteristic finding of Brenner tumor of the ovary on CT and MRI. PMID- 10667664 TI - Delineation of simulated vascular stenosis with Gd-DTPA-enhanced 3D gradient echo MR angiography: an experimental study. AB - PURPOSE: The purpose of this experimental study was to evaluate the influence of contrast material concentration and flow velocity on pulsatile flow in Gd-DTPA enhanced 3D gradient echo MR angiographic sequence. METHOD: In vivo flow experiments were performed in Plexiglas phantoms with artificial stenosis (50% stenotic ratio and 20 mm stenotic length) attached to a cardiac pump that generated physiological pulsatile flow similar to that of the bloodstream in a closed circuit. We used a steady-state gradient echo sequence with different TEs (6, 3, and 1.4 ms). A TR of 15 ms was used for all parameters. The concentration of Gd-DTPA varied from 0 to 2.0 mmol/L and flow velocities from 25 to 80 cm/s. We measured the degree of stenosis and length of stenosis in comparison with the actual values. RESULTS: The degree and length of stenosis on 3D gradient echo MR angiographic images were markedly influenced by the velocity of the flow and concentration of Gd-DTPA. The degree of stenosis was overestimated when the flow was fast or when the concentration of Gd-DTPA was low. When the concentration of Gd-DTPA was low, stenosis was elongated. These effects were less prominent on short TE (1.4 ms) sequence. CONCLUSION: The stenotic lesions were markedly overestimated on MR angiographic images obtained with Gd-DTPA-enhanced fast 3D gradient echo sequence. Spin dephasing can be compensated for almost entirely by a high concentration of Gd-DTPA and/or a short TE sequence. PMID- 10667665 TI - Intravenous leiomyomatosis. AB - We present two cases of intravenous leiomyomatosis with uterine leiomyoma or previous hysterectomy because of uterine leiomyoma in which MRI was characteristic. MRI showed a mass in the inferior vena cava and the heart. These MR findings are useful in diagnosing intravenous leiomyomatosis. PMID- 10667666 TI - Spared flow tract within the mural thrombus of an aortic aneurysm: its pathogenesis and clinical importance. AB - The purpose of this case report is to determine the unique pathogenesis of a "spared flow tract" through a thick mural thrombus of an aortic aneurysm mimicking the penetrating or dissecting tract of an impending or acute rupture of an abdominal aortic aneurysm (AAA) and to discuss its clinical importance. Three blood flow tracts (i.e., spared flow tracts) penetrating to aortic major branches (inferior mesenteric arteries in two and left renal artery in one) through thick mural thrombi of three aortic aneurysms were found on thin section spiral CT scans. Histopathological examination revealed that the tracts were formed by thrombi and partially covered with endothelial cells. In conclusion, spared flow tracts may be pathways continuing to the aortic major branches through thick mural thrombi of aortic aneurysms and are spared from thrombogenesis because of relatively high blood flows. Their pathogenesis is definitely different from penetrating or dissecting tracts within mural thrombi of ruptured AAAs. Spared flow tracts should not be misinterpreted as penetrating or dissecting tracts of impending or acute rupture. PMID- 10667667 TI - Acute pulmonary trunk dissection in a patient with primary pulmonary hypertension. AB - Spiral CT imaging findings including multiplanar reconstructions of an acute dissection of the pulmonary trunk in a 22-year-old female patient with primary pulmonary hypertension (PPH) are presented and discussed. PMID- 10667668 TI - Intracranial aneurysm on CTA: demonstration using a transparency volume-rendering technique. AB - We report an interesting transparency study using a volume-rendering technique applied to CT angiography in a patient with a sylvian aneurysm. On a single view, all the information required for the aneurysmal treatment could be analyzed. Comparison with maximum intensity projection and virtual endoscopy reconstructions was performed. PMID- 10667669 TI - Comparison of fMRI and intraoperative direct cortical stimulation in localization of receptive language areas. AB - PURPOSE: The purpose of this work was to compare the cortical localization of receptive speech using functional MRI (fMRI) and direct intraoperative electrical stimulation. METHOD: Three strongly right-handed patients with primary neoplasms of the left parasylvian region underwent fMRI while subjected to a passive listening task designed to activate receptive language areas. All three subjects then underwent awake intraoperative language mapping using direct electrical stimulation of the cortex. RESULTS: In all three subjects, similar, but nonidentical, cortical regions were identified as involved in receptive language function by fMRI and direct cortical stimulation mapping. CONCLUSION: fMRI provides excellent receptive language mapping, but its results must be interpreted with caution due to conceptual and technical differences from direct cortical stimulation mapping. PMID- 10667670 TI - Taste and smell phantoms revealed by brain functional MRI (fMRI). AB - PURPOSE: Our goal was to demonstrate the appearance of phantom tastes and smells (phantageusia and phantosmia, respectively) by use of functional MRI (fMRI) of the brain and to demonstrate the efficacy of drug treatment that inhibited both the subjective presence of these phantoms and the fMRI brain activation initiated by these phantoms. METHOD: Multislice FLASH MR or echo planar MR brain scans were obtained in two patients with phantageusia and phantosmia in response to memory of two tastants (salt and sweet); memory of two odors (banana and peppermint); actual smell of amyl acetate, menthone, and pyridine; and memory of phantom tastes and smells before and after treatment with thioridazine and haloperidol. Activation images were derived using correlation analysis, and ratios of brain area activated to total brain area were obtained. RESULTS: Prior to treatment, both patients experienced persistent birhinal and global oral obnoxious tastes and smells in the absence of any external stimulus. The fMRI response to memory of phantoms was activation in sensory-specific brain regions for taste and smell, respectively. fMRI activation was greater than for memory of any tastant or odorant or for actual smell of any odor. After treatment with thioridazine or haloperidol, which successfully inhibited each phantom in each patient, fMRI response to phantom memory was significantly inhibited and was significantly lower than for memory of any tastant or odorant or actual smell of any odorant. CONCLUSION: These results demonstrate that (a) phantom taste and smell can be revealed by fMRI brain activation, (b) brain activation in response to taste and smell phantoms is localized in sensory-specific brain regions for taste and smell, respectively, (c) brain activation in response to memory of each phantom initiated the greatest degree of activation we had previously measured, and (d) treatment with thioridazine or haloperidol inhibited both the presence of each phantom and its associated fMRI brain activation. This is the first study in which phantom tastes and smells have been demonstrated by an objective technique and treatment that inhibited the phantoms was characterized by objective inhibition of fMRI activation. These two patients represent a relatively common group that may be classified as having primary phantageusia and phantosmia distinct from those with phantoms or auras secondary to neurological, migrainous, psychiatric, or other causes. PMID- 10667671 TI - Cerebral perfusion MRI with arterial spin labeling technique at 0.5 Tesla. AB - PURPOSE: Our aim was to evaluate the feasibility of cerebral perfusion MRI using an arterial spin labeling technique at 0.5 T. METHOD: We performed perfusion imaging with a flow-sensitive alternating inversion recovery (FAIR) sequence in a total of 37 patients with cerebral infarction. RESULTS: FAIR perfusion images demonstrated areas of pathological perfusion corresponding (13 patients) or not corresponding (15 patients) to the infarcted area on MR images. Among 19 patients in whom comparison between FAIR perfusion imaging and regional cerebral blood flow single photon emission CT was available, the two studies correlated well in 15 patients. CONCLUSION: Our results indicate that the FAIR technique allows reliable cerebral perfusion imaging at 0.5 T. PMID- 10667672 TI - Analyzing functional brain images in a probabilistic atlas: a validation of subvolume thresholding. AB - PURPOSE: The development of structural probabilistic brain atlases provides the framework for new analytic methods capable of combining anatomic information with the statistical mapping of functional brain data. Approaches for statistical mapping that utilize information about the anatomic variability and registration errors of a population within the Talairach atlas space will enhance our understanding of the interplay between human brain structure and function. METHOD: We present a subvolume thresholding (SVT) method for analyzing positron emission tomography (PET) and single photon emission CT data and determining separately the statistical significance of the effects of motor stimulation on brain perfusion. Incorporation of a priori anatomical information into the functional SVT model is achieved by selecting a proper anatomically partitioned probabilistic atlas for the data. We use a general Gaussian random field model to account for the intrinsic differences in intensity distribution across brain regions related to the physiology of brain activation, attenuation effects, dead time, and other corrections in PET imaging and data reconstruction. RESULTS: H2(15)O PET scans were acquired from six normal subjects under two different activation paradigms: left-hand and right-hand finger-tracking task with visual stimulus. Regional region-of-interest and local (voxel) group differences between the left and right motor tasks were obtained using nonparametric stochastic variance estimates. As expected from our simple finger movement paradigm, significant activation (z = 6.7) was identified in the left motor cortex for the right movement task and significant activation (z = 6.3) for the left movement task in the right motor cortex. CONCLUSION: We propose, test, and validate a probabilistic SVT method for mapping statistical variability between groups in subtraction paradigm studies of functional brain data. This method incorporates knowledge of, and controls for, anatomic variability contained in modern human brain probabilistic atlases in functional statistical mapping of the brain. PMID- 10667673 TI - The identification of cerebral volume changes in treated growth hormone-deficient adults using serial 3D MR image processing. AB - PURPOSE: A pilot study to detect volume changes of cerebral structures in growth hormone (GH)-deficient adults treated with GH using serial 3D MR image processing and to assess need for segmentation prior to registration was conducted. METHOD: Volume MR scans of the brain were obtained in five patients and six control subjects. Patients were scanned before and after 3 and 6 months of therapy. Control subjects were scanned at the same intervals. A phantom was used to quantify scaling errors. Second and third volumes were aligned with the baseline by maximizing normalized mutual information and transformed using sinc interpolation. Registration was performed with and without brain segmentation and correction of scaling errors. Each registered, transformed image had the original subtracted, generating a difference image. Structural change and effects of segmentation and scaling error correction were assessed on original and difference images. The radiologists' ability to detect volume change was also assessed. RESULTS: Compared with control subjects, GH-treated subjects had an increase in cerebral volume and reduction in ventricular volume (p = 0.91 x 10( 3)). Scale correction and segmentation made no difference (p = 1 and p = 0.873). Structural changes were identified in the difference images but not in the original (p = 0.136). The radiologists detected changes >200 microm. CONCLUSION: GH treatment in deficient patients results in cerebral volume changes detectable by registration and subtraction of serial MR studies but not by standard assessment of images. This registration method did not require prior segmentation. PMID- 10667674 TI - Evaluation of 201T1 SPECT for predicting early treatment response in patients with squamous cell carcinoma of the extracranial head and neck treated with nonsurgical organ preservation therapy: initial results. AB - PURPOSE: The purpose of this work was to prospectively determine the ability of 210TI single photon emission CT (SPECT) to monitor treatment response in patients with head and neck squamous cell carcinoma (HNSCCA) treated with nonsurgical organ preservation. METHOD: Nine patients with HNSCCA underwent 201T1 SPECT before and 6 weeks after completion of nonsurgical organ preservation therapy. All cases were evaluated for uptake at the primary site before and after treatment. All tumors had abnormal radiotracer uptake on the pretreatment study. The posttreatment thallium studies were evaluated for uptake and correlated with local control at the primary site in all cases. RESULTS: All patients had abnormal thallium uptake on pretreatment studies. Of the nine patients, four cases were locally controlled by nonsurgical organ preservation therapy. All of these patients had no evidence of thallium uptake on posttreatment studies. Five cases failed treatment at the primary site. All five patients demonstrated abnormal radiotracer uptake at the primary site. CONCLUSION: Our initial results suggest that 201T1 SPECT may be an accurate technique for monitoring HNSCCA treated with nonsurgical organ preservation therapy. PMID- 10667675 TI - Augmentation procedures of the jaw in patients with inadequate bone for dental implants: radiographic appearance. AB - Dental implants, metallic posts surgically imbedded in the jaw to support dental prostheses, have provided an attractive alternative to standard removable dentures. Some patients, however, have insufficient bone to accommodate these implants. In this setting, a number of surgical procedures are available to augment the bone in the jaw. It is important for radiologists to be familiar with these procedures because the altered anatomy can be a source of confusion to the unwary. The objective, therefore, was to describe these procedures and their radiographic appearances. PMID- 10667676 TI - MR observations of postraumatic osteolysis of the distal clavicle after traumatic separation of the acromioclavicular joint. AB - PURPOSE: The purpose of this work was to characterize the MR features of post traumatic osteolysis of the distal clavicle in patients who have sustained a previous separation of the ipsilateral acromioclavicular (AC) joint. METHOD: We studied eight male patients (mean age 25 years) with intractable pain in the AC joint after sustaining a traumatic joint separation. With use of the Rockwood classification, the separations were classified as Type 1 in one patient, Type 2 in two patients, and Type 3 in five patients. The MR studies were evaluated for periarticular soft tissue swelling, cortical irregularity defined as thinning or absence of portions of the cortex in the acromial and clavicular articular surfaces, hypertrophic osseous changes, periostitis, bone marrow edema, periarticular cyst-like changes, and joint space widening exceeding 6 mm. Radiographs were evaluated independently of the MR studies. Osteolysis of the distal clavicle was confirmed pathologically in seven patients and with surgery in one patient. RESULTS: The incidence of osteolysis in patients who have had a previous AC joint separation was estimated to be approximately 6%. Observations on MRI included soft tissue swelling, bone marrow edema in the distal clavicle, and cortical irregularity associated with periarticular cyst-like erosions in eight patients, joint space widening in six patients, clavicular periostitis in three patients, and marrow edema in the cromion in five patients. Only one patient had osteophyte formation. Radiographic observations of periarticular soft tissue swelling, osteopenia of the distal clavicle, articular erosions, and joint space widening allowed diagnosis in only four patients prospectively. CONCLUSION: The MR features of posttraumatic osteolysis are characteristic of this process. We advocate the use of MRI in patients with chronic AC joint pain who have had a prior AC joint dislocation, particularly if follow-up radiographs are nonspecific, equivocal, or do not indicate the presence of secondary osteoarthritis. PMID- 10667678 TI - Metaphyseal undertubulation in gaucher disease: resolution at MRI in a patient undergoing enzyme replacement therapy. AB - Gaucher disease is a sphingolipid storage disorder that results in the accumulation of Gaucher cells within the reticuloendothelial system. The life span can be near normal in the most common form. Our case illustrates the resolution of the skeletal findings in Gaucher disease following enzyme replacement therapy. We also report the correlation of these findings with clinical improvement. PMID- 10667677 TI - CT of metal implants: reduction of artifacts using an extended CT scale technique. AB - PURPOSE: The purpose of this work was to use an extended CT scale technique (ECTS) to reduce artifacts due to metal implants and to optimize CT imaging parameters for metal implants using an experimental model. METHOD: Osteotomies were performed in 20 porcine femur specimens. One hundred cobalt-base screws and 24 steel plates were used for osteosynthesis in these specimens. Artificial lesions were produced in 50 screws, such as osteolysis near the screws (mimicking lysis due to infection, tumor, or loosening), displacement of the screws, as well as fractures of the screws. All specimens were examined using eight different CT protocols: four conventional (CCT) and four spiral (SCT) CT protocols with different milliampere-second values (130 and 480 mAs for CCT, 130 and 300 mAs for SCT), kilovolt potentials (120 and 140 kVp), and slice thicknesses (2 and 5 mm). The images were analyzed by three observers using a standard window (maximum window width 4,000 HU) and ECTS (maximum window width 40,000 HU). Receiver operating characteristic analysis was performed, and image quality was assessed according to a five level scale. RESULTS: Metal artifacts were significantly reduced using ECTS (p < 0.05). The highest diagnostic performance was obtained using ECTS with the thinnest slice thickness. Metal artifacts were more pronounced using SCT. In this experimental model, exposure dose and kilovolt potential had no significant impact on diagnostic performance (p > 0.05). CONCLUSION: ECTS improved imaging of metal implants. In this study, no significant effects of exposure dose and kilovolt potential were noted. Metal artifacts were more prominent using SCT than using CCT. PMID- 10667679 TI - MRI in sacral echinococcosis. PMID- 10667680 TI - Aunt Minnies's Corner. A case with radiologic findings so specific that no realistic differential diagnosis exists. PMID- 10667681 TI - The Stockholm Consensus Conference on quality specifications in laboratory medicine, 25-26 April 1999. PMID- 10667682 TI - Introduction: strategies to set global quality specifications in laboratory medicine. PMID- 10667683 TI - Determination and application of desirable analytical performance goals: the ISO/TC 212 approach. AB - Desirable analytical performance goals are needed to place clinical laboratories' quality management plan on a rational basis. While the most important basis of desirable analytical performance goals is medical needs, a well-established quality management plan must take into account economic and regulatory needs. There are many approaches that have been proposed for determining desirable analytical performance goals. The ISO/TC 212 approach for rationalizing the various approaches for determining desirable analytical performance goals, which is based on the ability to meet medical needs, is presented. PMID- 10667684 TI - The need for a system of quality standards for modern quality management. AB - The management of analytical quality depends on the careful evaluation of the imprecision (uncertainty) and inaccuracy (trueness) of laboratory methods and the application of statistical quality control procedures to detect medically important analytical errors that may occur during routine analysis. A system of quality standards is recommended to incorporate different types of requirements, such as clinical outcome criteria, analytical outcome criteria, and analytical performance criteria. For practical applications, all need to be translated into operating specifications for the imprecision, inaccuracy, control rules, and number of control measurements that are necessary to assure analytical quality during routine production of test results. PMID- 10667685 TI - General strategies to set quality specifications for reliability performance characteristics. AB - Many strategies have been promulgated for the setting of quality specifications in laboratory medicine. Based on the analysis of the effect of error on clinical decision making, general quality specifications for precision, bias, the allowable difference between two analytical methods, drugs, fixed limits for use in external quality assessment and reference methods seem best derived from components of biological variation. PMID- 10667686 TI - Current databases on biological variation: pros, cons and progress. AB - A database with reliable information to derive definitive analytical quality specifications for a large number of clinical laboratory tests was prepared in this work. This was achieved by comparing and correlating descriptive data and relevant observations with the biological variation information, an approach that had not been used in the previous efforts of this type. The material compiled in the database was obtained from published articles referenced in BIOS, CURRENT CONTENTS, EMBASE and MEDLINE using "biological variation & laboratory medicine" as key words, as well as books and doctoral theses provided by their authors. The database covers 316 quantities and reviews 191 articles, fewer than 10 of which had to be rejected. The within- and between-subject coefficients of variation and the subsequent desirable quality specifications for precision, bias and total error for all the quantities accepted are presented. Sex-related stratification of results was justified for only four quantities and, in these cases, quality specifications were derived from the group with lower within-subject variation. For certain quantities, biological variation in pathological states was higher than in the healthy state. In these cases, quality specifications were derived only from the healthy population (most stringent). Several quantities (particularly hormones) have been treated in very few articles and the results found are highly discrepant. Therefore, professionals in laboratory medicine should be strongly encouraged to study the quantities for which results are discrepant, the 90 quantities described in only one paper and the numerous quantities that have not been the subject of study. PMID- 10667687 TI - Series analyses and quality specifications required for monitoring over time. AB - General specifications of analytical goals for biochemical monitoring have been proposed based on biological within-subject variation. As a complement to this strategy for general global quality requirements there is also a need for other methods and approaches to set more optimal requirements in specific monitoring situations, i.e. application of systems and sensitivity analysis using (i) rules for propagation of errors (uncertainty) in simple algebraic/statistical transformations of laboratory results; (ii) biochemical/pathophysiological simulation models; and (iii) formalized descriptions of clinical classification and decision support processes. The possible gain in medical outcome by improving analytical and preanalytical quality should then be related to the often more important aspects of selection and combination of "tests", period and frequency of sampling/measurement, and the use of correct conceptual and computational models for transformation of data. PMID- 10667688 TI - Analytic bias specifications based on the analysis of effects on performance of medical guidelines. AB - Laboratory tests are key indicators for certain practice guidelines, and analytic bias can significantly alter the performance of these guidelines. Three clinical paradigms are described: serum cholesterol testing for risk assessment of cardiac disease, serum thyroid-stimulating hormone (TSH) measurement for the detection of hypothyroidism, and serum prostate-specific antigen (PSA) testing for prostate cancer risk assessment. Maximum tolerance limits for analytic bias are calculated by assessing the subgroup population fluctuations in the number of patients exceeding the guideline threshold values and limiting the analytic bias to one half of these fluctuations. Our calculated maximum bias limits are +/-1% for cholesterol and +/-6% for TSH and PSA. Our recommended +/-1% bias limit for cholesterol allows for a -6.5% to + 5.8% change in the number of patients designated as at risk for cardiac disease, whereas the +/-3% National Cholesterol Education Program limits permit a -18.4% to +16.7% variation. Similarly, our +/ 6% bias limits for TSH allow a -17.7% to +26.6% change in patients flagged for hypothyroidism, whereas the +/-10% bias values found with many commercial reagents permit a -28.2% to +49.2% variation in patient classification. Our +/-6% PSA bias limits correspond to changes from -14.2% to +11.4% in the number of men classified as at risk for prostate cancer. The +/-10% bias ranges for PSA correspond to -19.9% to +20.4% variation in patient classification. The larger tolerance limits of the CLIA-88 standards for proficiency testing would cause even wider variations in patient classifications. PMID- 10667689 TI - Quality specifications based on the uncertainty of measurement. AB - The result of a measurement needs to be accompanied by information on the error or uncertainty of the measurement. Either of two very closely related concepts is thus required to indicate the usefulness and relevance of a result for its purpose: decision making. This presentation will outline the difference between the concept of error and that of uncertainty and deal with procedures to estimate the latter. The use of a spreadsheet program will illustrate how the combined uncertainty is estimated in a relatively complex situation with a bearing on laboratory medicine. PMID- 10667690 TI - Quality specifications based on analysis of effects of performance on clinical decision-making. AB - The graphical model for evaluation of analytical requirements for bimodal distributions, based on probit transformation and calculation of false-positive and false-negative results for assumed random and systematic analytical errors, is presented in theory. It is concluded that the bimodal model is an excellent tool for evaluation of the effect of analytical quality and, therefore, useful for estimation of analytical quality specifications for quantities used in specific clinical strategies and situations as well as based on analysis of clinical outcome. PMID- 10667691 TI - Quality specifications for ordinal scale measurements with multiproperty (multiple) urine test strips. AB - Analytical quality specifications for ordinal scale measurements have not been presented so far. Criteria are suggested for multiproperty (multiple) urine test strips based on upper limits of healthy reference intervals, analytical performance and statistical tests applicable to ordinal scales. Trueness (accuracy) can be evaluated against an acceptable comparison method by applying sensitivity and specificity concepts, and defining a grey zone with a lower detection limit and an upper confirmation limit. Concordance (agreement) of two or more ordinal scale categories should be evaluated by subtracting random agreement, using Kappa statistics. Repeatability (precision) can be calculated for categorized results using binomial statistics. PMID- 10667692 TI - Setting process control limits for enzyme tests in serum. AB - With the advent of much more precise laboratory equipment, the use of performance based standard deviations for enzymes, e.g. those based on the precision of the laboratory during a period of "satisfactory performance", is not appropriate; it leads to overly rigid precision limits and unnecessary repeat assays, and is wasteful of resources. Testing for the commonly used serum enzymes does not require tight precision given how enzyme data are viewed by most clinicians. Other ways to set process control limits all have some arbitrary content; however, this need not prevent their use. Laboratorians must choose the approach that is appropriate for their laboratory and medical staff to meet medically perceived needs. PMID- 10667693 TI - Quality specifications derived from objective analyses based upon clinical needs. AB - The well-informed physician should be the best to judge the analytical quality needed for a specific clinical action. By presenting the physicians with well designed case histories, it is possible to extract the clinical knowledge pertinent to analytical quality. The clinicians are asked which changes in the concentration of an analyte will cause them to change actions concerning the patient. It must be possible to detect these changes with a certain probability and this presupposes a certain analytical quality which can then be calculated. It is important that a strict methodology both for constructing case histories and for calculation of the required analytical quality is used. PMID- 10667694 TI - Quality specifications for reference methods. AB - Reference methods are a key element to the objective of traceability in laboratory medicine. However, to serve this purpose adequately, minimum analytical quality specifications are required. Here, possible strategies for deriving such specifications are presented, being based on concepts developed before by a European Working Group. Distinction is made between "genuine requirements" for reference methods (direct calibration with primary reference materials; absence of sample-related effects) and "performance specifications" (limits for random, systematic and total error, the latter in association with the number of measurements). While the former requirements are considered as conditio sine-qua-non, the latter specifications should be variable, which means that they should be tailored to the specific application of the methods. In general, it is advocated to derive performance specifications for reference methods from desirable specifications of routine methods (analyte-related), although other models should not be ruled out beforehand. Further, it is recommended that reference laboratories make special efforts to demonstrate and maintain a uniform level of quality of reference methods. PMID- 10667695 TI - Development and use of analytical quality specifications in the in vitro diagnostics medical device industry. AB - Manufacturers of in vitro diagnostic (IVD) medical devices have become integral partners with their customers in determining the quality of laboratory results. Design controls imposed by ISO 9001 quality system standards and various regulations require manufacturers to implement a formal design process, which begins and ends with customer requirements. For IVD systems, this means that manufacturers must establish analytical quality specifications as part of their design input. This provides greater assurance that commercial products will satisfy customer requirements. In the case of quantitative IVD measurement systems, analytical quality specifications include total allowable uncertainty (bias, imprecision, non-specificity). The primary source of customer requirements is the laboratory-customer, who should have established analytical quality specifications based on the needs of its physician-clients. The total allowable uncertainty budget is allocated in the design process to the individual components of the system, such as reagents, instrumentation, calibrators and accessories, and to other factors such as operator, specimen and environmental interactions. Their performance must collectively meet the total allowable uncertainty specification when they are finally integrated into a measurement system. The design control model requires objective evidence that design specifications have been met (verification) and, finally, that the system will satisfy the needs of its intended users (validation). Compliance with the quality system standards is monitored through independent audits, government inspections and post-market surveillance. PMID- 10667696 TI - Preanalytical factors and their influence on analytical quality specifications. AB - Whereas analytical standards are described by established quality control criteria, no such standards exist for defining the quality of the preanalytical phase. A working group of the German Society for Clinical Chemistry and Laboratory Medicine has defined recommendations to describe the quality criteria for materials and processes used in the diagnostic process between patients and the analytical step. Thus, the quality of the sample may be defined regarding its adequacy and amount, as well as anticoagulants and stabilizers used. Timing of sampling, transport and storage involve criteria on sample stability, proper transport and preanalytical treatment. Moreover, sample identification, storage, and handling of interference and influence factors can be documented in quality manuals for the preanalytical phase. These possible variables have been discussed in five European expert meetings and recommendations published in national journals and presented in book form. PMID- 10667697 TI - Postanalytical factors and their influence on analytical quality specifications. AB - Quality thinking in medical laboratories is placed within the framework of the European Foundation for Quality Management Model which suggests the use of medical outcome indicators such as those developed by the Joint Commission on Accreditation of Healthcare Organizations. From here concepts were illustrated to improve the quality of the medical laboratory towards the data produced but moreover towards the information given, the knowledge acquired and the decision taken. Postanalytical factors were used to define analytical quality specifications. PMID- 10667698 TI - Analytical quality, performance indices and laboratory service. AB - Faulty data lead to suboptimal diagnostics and decision making unless the flaw is known and amenable to correction. However, pure noise (added analytical variance) has only minor effects on clinically appropriate indices of diagnostic performance. This fact is illustrated by an idealized screening programme, using the preventive benefit-to-cost ratio as the index of performance. Further, this article illustrates that the effect of an unnoticed source of noise is roughly just twice the effect that the same noise will have when its magnitude is known and clinical decision limits are adjusted accordingly. Owing to the small size of these effects, however, it may be profitable to spend resources on other aspects of good laboratory service, such as timeliness, documentation and interpretative support. PMID- 10667699 TI - Quality specifications of clinical laboratory procedures: developing country needs. AB - In developing countries laboratory measurements are made in specimens of populations that are usually of different genetic origin and live in a different environment, under working conditions that differ from those of industrialized countries. The setting of quality specifications of laboratory measurements must take into account such differences to improve the quality of laboratory services in these countries. Industry and professional organizations should be aware of the differences, otherwise their recommendations may counteract efforts to improve laboratory services in developing countries. PMID- 10667700 TI - Effect of legislation (CLIA'88) on setting quality specifications for US laboratories. AB - The Clinical Laboratory Improvement Amendments of 1988 (CLIA'88) mandated, in response to concern over the perceived quality of clinical laboratory testing, universal regulation for all testing sites in the USA, including previously unregulated sites in physician offices. The intent of CLIA'88 is to ensure quality of testing through a combination of total quality management and mandated minimum quality practices. CLIA also defines, intentionally or unintentionally, through its proficiency testing requirements, intralaboratory performance standards. Meeting these requirements has been a prime motivator in improving laboratory performance. Seven years after the implementation of CLIA'88, the percentage of laboratories passing proficiency testing has increased and most laboratories have implemented quality practices. PMID- 10667701 TI - Effect of current and forthcoming European legislation and standardization on the setting of quality specifications by laboratories. AB - A network of national and international guidelines and directives developed in the last few decades by various bodies will lead to a new concept of total quality for medical laboratory services comprising legislative regulations on national and international levels, standardizations backed up by legislation and recommendations of professional societies. One example is the IVD Directive of the European Community. It will not only stimulate accreditation in the field of laboratory medicine, but also necessitate numerous standardization activities which are presently co-ordinated by the European Committee for Standardization (CEN). Another standardization example is the development of quality management systems, mainly by ISO. The ISO 9000 series has become the most successful family of standards world-wide. Meanwhile, specific standards for the needs of laboratories (ISO 17025), and in particular of medical laboratories (ISO 15189), are being worked out. A new trend to develop quality management systems towards total quality management systems can be observed including additional aspects such as economic and quality interests of society, customers and owners of laboratories. The goal of all activities is to create a network of confidence which provides some guarantee to the clients, i.e. the physicians and their patients, that they will receive a high-quality medical laboratory service. PMID- 10667702 TI - Effect of accreditation schemes on the setting of quality specifications by laboratories. AB - In the future, there will be a universal standard for quality management in medical laboratories: ISO 15189. This standard follows the basic principles of ISO 17025, the general standard for test laboratories, but also adds several specific aspects. A comparison between these standards is given. The language of ISO 15189 is designed to be understood by medical laboratory professionals. As this standard is applicable to all medical laboratory fields, requirements are given in general terms requiring the laboratory to implement them correctly. Because it is essential that information provided by laboratory results is useful for healthcare, the requirements covered by ISO 15189 are compared with those needed for providing good medical laboratory services. The capabilities of the personnel at the laboratory clinic interface are the most difficult to assess and evaluate in an adequate quality management system. PMID- 10667703 TI - Setting quality specifications for the future with newer approaches to defining uncertainty in laboratory medicine. AB - When describing the performance of procedures and the reliability of their results, ISO terminology should be used. Results should be universally comparable and this requires metrological traceability. The concomitant uncertainty (inversely) indicating reliability should be obtained in a universal and transparent fashion, and should be combinable. Therefore, the approach of the "Guide to the expression of uncertainty in measurement", leading to a result without known bias and a combined standard uncertainty, has advantages over the allowable total error concept, incorporating procedural bias. PMID- 10667704 TI - The beginning of the artificial eye program. PMID- 10667705 TI - Artificial vision for the blind by connecting a television camera to the visual cortex. AB - Blindness is more feared by the public than any ailment with the exception of cancer and AIDS. We report the development of the first visual prosthesis providing useful "artificial vision" to a blind volunteer by connecting a digital video camera, computer, and associated electronics to the visual cortex of his brain. This device has been the objective of a development effort begun by our group in 1968 and represents realization of the prediction of an artificial vision system made by Benjamin Franklin in his report on the "kite and key" experiment, with which he discovered electricity in 1751. PMID- 10667706 TI - Peter Robert Uldall 1935-1995: a personal tribute. PMID- 10667708 TI - FDA reform legislation and its effect on clinical trials. PMID- 10667707 TI - Home hemodialysis: a crazy idea in 1963: a memoir. AB - In 1963, one of the abstracts we submitted for the Ninth ASAIO Congress entitled "Hemodialysis at Home: Utilizing Domestic Electric Washing Machine" by Y. Nose and J. Mikami was rejected. At that time, leading ASAIO nephrologists, including W. J. Kolff, B. H. Scribner, and J. P. Merrill, did not favor patients dialyzing at home in spite of it being the only means of keeping renal failure patients alive. Also at that time, many patients died because of the unavailability of hemodialysis in hospitals. However, in 1965 both Dr. Scribner's group in Seattle and Dr. Merrill's group in Boston initiated a home hemodialysis program. This program was further expanded in many centers, including Dr. Kolff's at Cleveland Clinic. Later, home hemodialysis was proven a safe, effective, yet inexpensive method of treating chronic renal insufficiency patients. PMID- 10667709 TI - Conduct of device clinical trials: the need for collaboration in response to a changing health care environment. AB - Prior to initiating a clinical trial, many issues need to be discussed to insure an adequate understanding of the investigators' and sponsors' limitations, capabilities, and expectations. Many of these issues are addressed in contracts and agreements, the formats for which have evolved over many years. In these documents, issues such as publication rights, access to results, confidentiality, conditions for termination, compensation, and indemnification are covered. However, other essential, although noncontractual issues that reflect upon trends in and the climate of the health care system, need to be discussed. Issues such as the development of collaborative cost-effectiveness studies, public relations plans, content of a cost-benefit analysis, results of other trials, reimbursement strategies, and the development of resources that contribute to the ease, safety, and efficacy of device use. With the increasing expense, oversight, and administrative burden for the conduct and participation in clinical trials, there is a correspondingly greater need for sponsor and investigator communication, collaboration, and risk and cost sharing to ensure the successful outcome of clinical trials today. PMID- 10667710 TI - Clinical trial: a clinician's perspective. AB - As a result of my participation in several clinical trials at The University of Iowa, I have now redrawn what I perceive to be the algorithm for conduct of a clinical trial. Of note, I have excluded the development of the new device, in vitro device testing, in vivo animal experimentation, clinical protocol development, granting of an Investigational Device Exemption, and the process by which clinical sites are selected. Following my algorithm I have made no mention of the commercialization of an experimental device, including the Pre-Market Approval process and FDA panel meetings. In summary, I believe there are a variety of reasons why a principal investigator would choose to participate in a clinical trial. With tongue in cheek, I would offer fame and fortune as two possible motivating factors. Obviously, both are double-edged swords. Should a trial not turn out as hoped, celebrity can instantly turn to notoriety. With respect to fortune, administrative and clinical activity performed by the principal investigator on behalf of the clinical trial does not generate income, and, in fact, can place a considerable financial drain on the investigator's department. I believe that investigators are strongly driven by intellectual curiosity, but it is important to maintain perspective when participating in a clinical trial. The real reason we seek clinical trial site designation is to allow us to offer new, innovative therapeutic modalities to our patients. By so doing, it is imperative that we conduct the clinical trial in as thoughtful and as safe a manner as possible, with our patients' well being always foremost in our minds. PMID- 10667711 TI - The conduct of clinical trials: a Food and Drug Agency perspective. AB - The conduct of a clinical trial required by FDA regulations should be executed in conformity with a protocol developed as a cooperative endeavor involving the Agency, the sponsor requiring device approval for marketing, and the investigators. This is the process most likely to assure a scientifically satisfactory study design that will address the primary concerns of all the parties. More importantly, timely access to safe and effective technologic advances will be made available to the health provider community based on the best scientific evidence. PMID- 10667712 TI - Clinical trials: a sponsor's perspective: overview. PMID- 10667713 TI - Bacterial components inhibit fibroblast proliferation in vitro. AB - Perigraft fluid from Staphylococcus epidermidis infected grafts in a mouse model significantly inhibits fibroblast proliferation (60-98% at 7 and 28 days), compared with perigraft fluid from sterile grafts. The fibroblast inhibitor was trypsin-heat resistant and dependent primarily upon the bacteria, not the host proinflammatory mediators or the vascular graft biomaterial. We tested the inhibitory properties of S. epidermidis strains RP62A (slime producer) and RP62NA (nonslime producer) and Staphylococcus aureus strain 502a, using an in vitro tritiated thymidine murine fibroblast (ATCC CCL-12) proliferation assay. Whole killed bacteria, disrupted bacteria (live and killed), bacterial supernatants, and purified cell wall products (peptidoglycan, teichoic acid, and lipoteichoic acid from disrupted bacteria) were studied. Significant fibroblast inhibition occurred for all three bacterial strains with disrupted bacteria (live or killed) and cell free bacteria derived supernatants. The fibroblast inhibitor from disrupted slime producing S. epidermidis was trypsin-heat resistant. The fibroblast inhibitor from disrupted S. aureus and supernatants for all three bacterial strains at 1 x 10(7) were trypsin-heat sensitive. Fibroblast inhibition was not dependent upon bacterial viability and not mediated by bacterial cell wall products. In conclusion, components of slime and nonslime producing S. epidermidis and S. aureus inhibit fibroblast proliferation. PMID- 10667714 TI - A siliconized hollow fiber membrane oxygenator. AB - Most membrane oxygenators are built with microporous fibers known for plasma leakage in long-term use such as extracorporeal life support or extracorporeal membrane oxygenation. The current study was designed to evaluate the Quadrox oxygenator in which the fibers have been coated with silicone (Jostra). Six calves (mean weight, 62 +/- 4 kg) were connected to cardiopulmonary bypass (CPB) by jugular venous and carotid arterial cannulation, with a mean flow rate of 3 L/min for 6 hours. They were randomly assigned to a standard Quadrox oxygenator (standard group, n = 3) or a siliconized Quadrox oxygenator (silicone group, n = 3). After 7 days, the animals were sacrificed. A standard battery of blood samples was taken before bypass, after mixing for 10 minutes, and after 1, 2, 5, and 6 hours of perfusion. Analysis of variance was used for repeated measurements. Total oxygen transfer and carbon dioxide transfer did not differ between groups (p = 0.5 for comparison). Blood trauma, evaluated by plasma hemoglobin (Hb), did not detect any significant hemolysis in either group. Thrombocyte and white blood cell count profiles in both groups were parallel and without significant differences (p = 0.1 and 0.6, respectively). At the end of testing no clot deposition was found in the oxygenator. At postmortem, there were no signs of peripheral emboli. The results of this study suggest that this silicone coating of hollow fibers allows for good gas transfer, while preserving all the mechanical advantages of a conventional hollow fiber oxygenator. PMID- 10667715 TI - Hemodynamic effects of attachment modes and device design of a thoracic artificial lung. AB - A thoracic artificial lung (TAL) was designed to treat respiratory insufficiency, acting as a temporary assist device in acute cases or as a bridge to transplant in chronic cases. We developed a computational model of the pulmonary circulatory system with the TAL inserted. The model was employed to investigate the effects of parameter values and flow distributions on power generated by the right ventricle, pulsatility in the pulmonary system, inlet flow to the left atrium, and input impedance. The ratio of right ventricle (RV) power to cardiac output ranges between 0.05 and 0.10 W/(L/min) from implantation configurations of low impedance to those of high impedance, with a control value of 0.04 W/(L/min). Addition of an inlet compliance to the TAL reduces right heart power (RHP) and impedance. A compliant TAL housing reduces flow pulsatility in the fiber bundle, thus affecting oxygen transfer rates. An elevated bundle resistance reduces flow pulsatility in the bundle, but at the expense of increased right heart power. The hybrid implantation mode, with inflow to the TAL from the proximal pulmonary artery (PA), outflow branches to the distal PA and the left atrium (LA), a band around the PA between the two anastomoses, and a band around the outlet graft to the LA, is the best compromise between hemodynamic performance and preservation of some portion of the nonpulmonary functions of the natural lungs. PMID- 10667716 TI - Morphologic studies of hepatocytes entrapped in hollow fibers of a bioartificial liver. AB - A bioartificial liver cartridge was prepared by inoculating porcine hepatocytes into the inner space of hollow fibers of a hemodialyzer. The hepatocytes formed rod shaped cell aggregates during in vitro perfusion culture within 1 day. Morphologic examination was carried out on the aggregates by optical and electron microscopy. Each hepatocyte was in direct contact with adjacent cells and a bile canaliculus-like structure was occasionally seen between hepatocytes. High magnification observation showed that the canaliculus was separated from the remainder of the intercellular space by a tight junction. These facts suggest that the hepatocytes formed functionally associated cell aggregates with a compact morphology not unlike hepatocyte spheroids. These structures were well maintained for 7 days in culture, and then the amorphous area in the aggregates and the nonviable cell number increased with lengthening culture period. The bioartificial liver maintained the ability to metabolize lidocaine, ammonia, and galactose for 7 days and then deteriorated with time. PMID- 10667717 TI - Effects of recombinant human hepatocyte growth factor on the proliferation and function of porcine hepatocytes. AB - A porcine hepatocyte based bioartificial liver (BAL) is still insufficient to replace liver transplantation. In this experiment, to strengthen the performance of a BAL, the effect of human recombinant hepatocyte growth factor (rhHGF) on the proliferation and function of xenogeneic porcine hepatocytes was studied. Isolated porcine hepatocytes were seeded at various densities (5 x 10(3) to 8 x 10(4) cells/well) on a collagen coated 96 well plate in Dulbecco's modified Eagle's medium (DMEM) with 10% FCS. After 4 hours, the medium was changed to DMEM with added insulin and dexamethasone. Subsequently, rhHGF was added at various concentrations (0, 0.625, 1.25, 2.5, 5, 10, 20, 40 ng/ml) and cultured for an additional 24, 48, and 72 hours, respectively. The proliferation of porcine hepatocytes in response to rhHGF reached a plateau at 2.5 ng/ml at 24 hours and subsequently decreased. The levels of porcine albumin vs protein present in the supernatant increased when cultured at high cell density. In conclusion, rhHGF was found to stimulate proliferation of porcine hepatocytes at low cell density and low concentration. rhHGF can also increase albumin synthesis at higher cell density, thus indicating its potential use in a more satisfactory porcine hepatocyte based BAL. PMID- 10667718 TI - I: Negative effect of cold ischemia on initial renal function. AB - Correlation between post-transplant function and exposure to cold ischemia (CI) during preservation has been reported. We attempted to identify the effect of CI on renal function using exsanguinous metabolic support (EMS) technology, to eliminate effects of reperfusion complications. Small bovine kidneys were used to evaluate 4 vs. 24 hours of CI, after warm ischemic (WI) exposure of <15, 30 or 60 minutes. After CI, kidneys were warm perfused (30 degrees C to 32 degrees C) ex vivo using EMS technology. Restored renal metabolism and function were quantified by oxygen consumption, urine production, glomerular filtration rate (GFR), and hemodynamic characteristics. The results demonstrate a CI-associated lag phase in the restoration of metabolism, in which the longer cold-preserved kidneys exhibit a lower initial rate of oxygen consumption. However, after 3 hours of EMS perfusion there was no significant difference in the O2 consumed, urine flow, GFR, perfusion flow, or pressure between the kidneys stored for 4 or 24 hours. An initial reduction in metabolism after longer CI may influence the severity of actual reperfusion injury during transplantation. Therefore, these results provide preliminary evidence suggesting that an acellular warm temperature reperfusion ex vivo may enhance restoration of cellular metabolism and minimize damage from the cold seen upon actual reperfusion. PMID- 10667719 TI - II: Ex vivo viability testing of kidneys after postmortem warm ischemia. AB - Future approaches to expand the organ donor pool with marginal and nonheartbeating donors, will be dependent upon prospective organ evaluation. Restoration of metabolism by preservation at warmer temperatures could potentially provide the window for such evaluation. Using a small bovine model, kidneys were subjected to either < 15, < 30 or < 60 minutes of warm ischemia (WI) followed by cold ischemia (CI) in ViaSpan. After WI and CI, kidneys were transitioned to a warm temperature perfusion (30 degrees C to 32 degrees C) using exsanguinous metabolic support (EMS) technology. Restored renal metabolism and function was assessed by oxygen consumption, glucose consumption, urine production, glomerular filtration rate, and hemodynamic characteristics. The results of this study suggest that it is feasible to distinguish viable from nonviable organs ex vivo by assessing renal metabolism and function during warm preservation using EMS technology. PMID- 10667720 TI - Variable flow Doppler for hemodialysis access evaluation: theory and clinical feasibility. AB - Access thrombosis remains an enormous problem for patients on hemodialysis. Current evidence suggests that decreasing access blood flow rate is an important predictor of future access thrombosis and failure. This article describes a method for determining access volume flow and detecting access pathology. The Doppler ultrasound signal downstream from the arterial needle as a function of the variable hemodialysis blood pump flow rate, is used to determine access blood flow. By using this variable flow (VF) Doppler technique compared with duplex volume flow estimates measured in 18 accesses (16 patients with 12 polytetrafluorethylene [PTFE] grafts and 6 autogenous fistulas), the results showed a correlation of 0.83 (p < 0.0001) between these methods. In grafts with lower blood flow rates, aberrant flow patterns were observed, including stagnant or reversed flow during diastole while forward flow was maintained during systole. When reversed diastolic flow was severe, it was accompanied by access recirculation. In conclusion, we report the theory and clinical feasibility of determining access blood flow by using a VF Doppler technique. Measurements are made without the need to determine the access cross sectional area required for duplex volume flow calculations and without the need to reverse the lines required for various indicator dilution techniques. Important information is also obtained about aberrant flow patterns in patients at risk of access failure. PMID- 10667721 TI - A microdomain-structured synthetic high-flux hollow-fiber membrane for renal replacement therapy. AB - A prospective crossover clinical study evaluated solute removal and biocompatibility of a tailored, hydrophobic-hydrophilic microdomain structure produced from a blend of polyamide, polyarylethersulfone, and polyvinylpyrrolidone (Polyflux) compared with Fresenius Polysulfone in dialyzers of similar surface area. The clearance of small molecules (urea, creatinine, and phosphate) for both membranes was comparable. Plasma levels of beta2 microglobulin were reduced at the end of treatment with both membranes (49.8% of pretreatment values for Polyflux; 45.9%, Fresenius Polysulfone) and was associated with the recovery of 207 +/- 84 mg of beta2 microglobulin from the dialyzing fluid for Polyflux compared with 147 +/- 29 for Fresenius Polysulfone (p = 0.12). The dialyzing fluid also contained 7,758 mg of protein when using Polyflux compared with 7,793 mg using Fresenius Polysulfone (standard error of difference for any pair was 511 mg). No albumin was present in the dialysis fluid for either membrane. Neutropenia, platelet adhesion to the membrane, and complement activation characterized by C3a, C5a, and SC5b-9 generation were slight and independent of membrane type. Membrane thrombus generating potential measured by thrombin:antithrombin III were also similar. These results indicate that the tailored, hydrophobic-hydrophilic microdomain structure of the membrane results in a favorable biocompatibility profile and clinically acceptable solute removal similar to the widely used Fresenius Polysulfone membrane. PMID- 10667722 TI - Dependence of peritoneal clearances on body size in continuous ambulatory peritoneal dialysis: effect of the normalizing size indicator. AB - In peritoneal dialysis (PD), small solute clearances are normalized by body water (V) and body surface area (BSA). The purpose of this study was to identify if V or BSA produced stronger associations between body size and normalized clearances. We studied the relationship between four size indicators (V, BSA, height, and weight) and either peritoneal urea clearance normalized to V (Kt/V(ur)) and BSA (C(ur)) or creatinine clearance normalized to V (Kt/V(cr)) and BSA (C(cr)). A total of 613 clearance studies were performed in subjects on continuous ambulatory peritoneal dialysis (CAPD) with four daily exchanges and a 2 L fill volume. As size increased, the normalized peritoneal clearances decreased in a nonlinear fashion (regression: y = b0 + b1x(-1), where x is a size indicator and y is a normalized clearance). Significant (p < 0.001) negative correlations were found between each normalized clearance and each size indicator. However, in each case, the correlation was higher when V, rather than BSA, was used. For example, BSA correlated more closely with K/V(ur)(-0.660) than C(ur)(-0.556), and also with Kt/V(cr)(-0.579) than C(cr)(-0.446). Normalized clearances are smaller in large subjects on CAPD because one mathematic determinant of the clearance, the drain volume (Dv) normalized by V (Dv/V) or BSA (DV/BSA), decreases as size increases. The relationship between Dv/V or Dv/BSA and the size indicators was studied by the same nonlinear regression model. The correlations of the size indicators with Dv/V were also consistently higher than the corresponding correlations with Dv/BSA. In subjects who were on the same PD schedule, the dependence of clearances on size was consistently higher when V, rather than BSA, was the normalizing parameter. Because prescription of the dose of PD is based on body size, there is a practical advantage by using V as the sole normalizing parameter for both urea and creatinine clearance. PMID- 10667723 TI - Simulation study of the intercompartmental fluid shifts during hemodialysis. AB - Hypotension is the most frequent complication during hemodialysis. An important cause of hypotension is a decrease in the intravascular volume. In addition, a decrease in plasma osmolality may be a contributing factor. Modeling of sodium and ultrafiltration (UF) may help in the understanding of underlying relationships. We therefore simulated, in a mathematical model, the intercompartmental fluid shifts during standard hemodialysis (SHD), diffusive hemodialysis (DHD), and isolated ultrafiltration (IU). We analyzed the relative theoretical effect of hydration status, dialysate sodium concentration, the initial plasma concentrations of sodium and urea, and tissue permeation to solutes on the magnitude and direction of intracellular and intravascular volume changes. This theoretical analysis shows that the transcellular fluid shifts taking place during hemodialysis treatment are, to a great part, due to inhomogeneous distribution of regional blood flow and tissue fluid volumes. During hemodialysis treatment, the cellular fluid shifts in tissue groups with relatively high perfusion and small volume occur from the intra- to the extracellular spaces. However, the fluid shift in tissue groups with a low perfusion and large volume takes place in the opposite direction. The UF volume and rates, and the size of the sodium (Na+) gradient between the dialysate and blood side of the dialyzer membrane are the most important factors influencing the fluid shifts. Higher UF volumes and flow rates cause an increasing decline in the plasma volume in both SHD and IU. High dialysate sodium concentration (150 mEq L(-1)) helps plasma refilling slightly when compared with a normal dialysate sodium concentration (140 mEq L(-1)). However, a high dialysate sodium concentration is associated with a high plasma sodium rebound, which in turn may lead to interdialytic water intake resulting from thirst and may cause increased weight gain and hypertension. PMID- 10667724 TI - Solute removal characteristics of continuous recirculating peritoneal dialysis in experimental and clinical studies. AB - Continuous recirculating peritoneal dialysis (CRPD) was introduced to enhance solute removal efficiency in conventional peritoneal dialysis (PD) therapies such as continuous ambulatory peritoneal dialysis (CAPD). In CRPD, a portion of the dwell dialysate in the patient's peritoneal cavity is drained through a double lumen catheter and purified by an extracorporeal dialyzer. In this study, solute removal characteristics and safety of CRPD are examined in ex vivo and clinical studies. Recirculation dialysis experiments using nine dogs (13.6 +/- 2.5 kg of body weight) were carried out for 240 min in the ex vivo study, whereas another seven dogs (12.1 +/- 2.8 kg) received conventional peritoneal dialysis (CPD) (120 min dwelling x 2) and six additional dogs (11.9 +/- 2.7 kg) received a Tidal PD (20 min dwelling x 12; 50% of tidal volume ratio) as controls. The ex vivo study revealed that CRPD has a higher efficiency for solute removal than CPD and is equivalent to Tidal PD. In the BUN reduction rate, the 19.4 +/- 5.5% in 240 min CRPD (n = 9) was significantly higher (p < 0.05) than the 3.5 +/- 3.6% in 240 min CPD (n = 7) and equivalent to the 17.3 +/- 4.7% in 240 min Tidal PD (n = 6). Continuous recirculating peritoneal dialysis maintained a low UN level in the peritoneal cavity due to dialysis with an extracorporeal dialyzer. This tendency was also seen in creatinine removal. In the clinical study, CRPD (n = 10) and CPD (n = 5) treatments were used in three renal failure patients. Higher solute removal efficiency was shown in CRPD than in CPD treatments, and the urea peritoneal clearance was 14.1 +/- 4.4 ml/min in CRPD (n = 10), significantly higher (p < 0.05) than the 7.3 +/- 2.1 ml/min in CPD (n = 5). No fibrin formation occurred during CRPD treatments. PMID- 10667725 TI - Internal mammary artery harvesting using the harmonic scalpel. AB - Internal mammary artery (IMA) harvesting using the harmonic scalpel (HS) was recently introduced. We studied 541 IMAs harvested by the same surgeon through a standard median sternotomy in 472 coronary bypass patients; 252 (47%) with the HS, while 289 (53%) were with electrocautery (EC). Patient demographics included mean ages: 67 years HS vs. 65 years EC (p = NS); male:female ratio: 3:1; and insulin dependent diabetes mellitus (IDDM): 11% HS vs. 12.5% EC (NS). Mean ultrasonic IMA flow at a mean SBP of 70 mm Hg in 10 consecutive patients of each group were: preharvest, HS 11.9 +/- 2.3 ml/min vs. EC 8.5 +/- 1.6 ml/min (p = 0.256); postharvest, HS 35.7 +/- 10.7 ml/min vs. EC 22 +/- 2.9 ml/min (0.235); and postcardiopulmonary bypass (post-CPB), HS 47.8 +/- 6.2 ml/min vs. EC 41.7 +/- 2.5 ml/min (0.381). Histologic samples of 50 consecutive IMAs showed no evidence of vessel injury in either group. Clinical results revealed postoperative bleeding in 6/217 (2.7%) HS vs. 7/255 (2.7%) EC (p = 0.783), none attributed to bleeding from the IMA; phrenic paresis: 0/217 in HS but 1/255 (0.4%) in EC (p = 0.960); sternal wound infection: 5/217 (2.3%) HS vs. 6/255 (2.4%) EC (p = 0.787); postoperative IABP: 6/217 (2.7%) HS vs. 5/255 (2%) EC (p = 0.859); mortality: 2/217 (0.9%) HS vs. 2/255 (0.8%) EC (p = 0.710). Hemodynamic, histologic, and clinical results were comparable in both groups. The authors believe the HS is safe and effective for IMA harvesting. PMID- 10667726 TI - Monitoring regional cerebral oxygen saturation using near-infrared spectroscopy during pulsatile hypothermic cardiopulmonary bypass in a neonatal piglet model. AB - Impairment of cerebral oxygenation in neonates and infants after hypothermic nonpulsatile cardiopulmonary bypass (CPB) support is well documented. The objectives of this study were: 1) using a neonatal piglet model to continuously monitor the regional cerebral oxygen saturation (rSO2) by near-infrared spectroscopy during pulsatile hypothermic CPB; and 2) to quantify the pulsatile flow in terms of energy equivalent pressure (EEP). After initiation of CPB, all piglets (n = 5) were subjected to 15 minutes of core cooling, reducing the rectal temperature to 25 degrees C, followed by 60 minutes of hypothermic CPB, then 10 minutes of cold reperfusion, and 30 minutes of rewarming. During CPB, mean arterial pressures (MAPs) and pump flow rates were maintained at 40-45 mm Hg and 150 ml/kg/min, respectively. During normothermic CPB, the rSO2 was significantly increased, compared with the pre-CPB level (56.8 +/- 5.2% vs. 41.8 +/- 5.5%, p < 0.01). At the end of cooling, the rSO2 level was 76.8 +/- 8.6% (p < 0.001 vs. pre CPB). After 60 minutes of hypothermic CPB and 30 minutes of rewarming, the rSO2 level was decreased to 38.6 +/- 4.2%, which was not significantly different compared with the pre-CPB level. The average increase in pressure (from MAP to EEP) was 5 +/- 1%, and the average increase in extracorporeal circuit pressure (from ECCP to EEP) was 13 +/- 2%. This extra pressure may help to provide better regional cerebral oxygen saturation. During pulsatile CPB, there was no rSO2 deficiency in this high flow model. Near-infrared spectroscopy responded well to changes in rSO2 during different stages of these experiments and might be a helpful tool for intraoperative monitoring. PMID- 10667727 TI - Evaluation of biodegradable, three-dimensional matrices for tissue engineering of heart valves. AB - A crucial factor in tissue engineering of heart valves is the type of scaffold material. In the following study, we tested three different biodegradable scaffold materials, polyglycolic acid (PGA), polyhydroxyalkanoate (PHA), and poly 4-hydroxybutyrate (P4HB), as scaffolds for tissue engineering of heart valves. We modified PHA and P4HB by a salt leaching technique to create a porous matrix. We constructed trileaflet heart valve scaffolds from each polymer and tested them in a pulsatile flow bioreactor. In addition, we evaluated the cell attachment to our polymers by creating four tubes of each material (length equals 4 cm; inner diameter, 0.5 cm), seeding each sample with 8,000,000 ovine vascular cells, and incubating the cell-polymer construct for 8 days (37 degrees C and 5% CO2). The seeded vascular constructs were exposed to continuous flow for 1 hour. Analysis of samples included DNA assay before and after flow exposure, 4-hydroxyproline assay, and environmental scanning electron microscopy (ESEM). We fabricated trileaflet heart valve scaffolds from porous PHA and porous P4HB, which opened and closed synchronously in a pulsatile bioreactor. It was not possible to create a functional trileaflet heart valve scaffold from PGA. After seeding and incubating the PGA-, PHA-, and P4HB-tubes, there were significantly (p < 0.001) more cells on PGA compared with PHA and P4HB. There were no significant differences among the materials after flow exposure, but there was a significantly higher collagen content (p < 0.017) on the PGA samples compared with P4HB and PHA. Cell attachment and collagen content was significantly higher on PGA samples compared with PHA and P4HB. However, PHA and P4HB also demonstrate a considerable amount of cell attachment and collagen development and share the major advantage that both materials are thermoplastic, making it possible to mold them into the shape of a functional scaffold for tissue engineering of heart valves. PMID- 10667728 TI - The effect of diastolic stiffness on ventricular function after partial ventriculectomy: a finite element simulation. AB - Partial ventriculectomy (PV) has been proposed by Batista and colleagues to improve cardiac function in patients with dilated cardiomyopathy (DCM); however, results have been mixed. We tested the hypothesis that preoperative diastolic function affects the stroke volume/end-diastolic pressure (Starling) relationship after PV. A previously described finite element simulation of DCM and PV was used. Diastole and end systole were represented by separate elastic finite element models with different unloaded shapes and nonlinear material properties. Left ventricular (LV) end-systolic elastance (E(ES)), diastolic compliance (DC), and Starling relationships were calculated. DC was varied by changing Ogden material property alpha(i) from 12 (compliant) to 20 (stiff). PV was simulated at 20% LV mass reduction. The slope of the Starling relationship increased from 1.82 to 1.21 as alpha(i) increased from 12 to 20. Partial ventriculectomy increased the Starling relationship in each case from 1.34 to 1.01 respectively. However, the net result in each case is a decrement in the Starling relationship with resection, and the smallest decrement was associated with the highest diastolic stiffness (alpha(i) = 20). Partial ventriculectomy depressed the Starling relationship for all values of diastolic compliance. It is expected that patients with a higher diastolic stiffness should do better. PMID- 10667729 TI - Terumo implantable left ventricular assist system: results of long-term animal study. AB - The research group of Terumo Corporation, NTN Corporation, and the Setsunan University have been developing an implantable left ventricular assist system (T ILVAS) featuring a centrifugal blood pump with a magnetically suspended impeller (MSCP). The present study describes results of chronic animal experiments using the MSCP. The MSCP has been tested ex vivo and in vivo in 6 sheep as a left heart bypass between the left ventricular apex and descending aorta. Ex vivo chronic sheep experiments using Model I demonstrated long-term durability, nonthrombogenicity, low hemolysis (<6 mg/dl), and excellent stability of the magnetic bearing with long-term survival for up to 864 days. Average pump flow rate was 4 L/min at a fixed rotational speed of 2000 rpm. Power spectral analyses of heart rate, aortic pressure, and blood temperature maintained normal 1/f fluctuation during the study. The retrieved pump was completely free from thrombus formation and there was no evidence of infarct in major organs. The implantable Model II was evaluated ex vivo in two sheep and intra-thoracically implanted in a sheep. These experiments were terminated at 70, 79, and 17 days due to blood leakage through the connector system within the housing. No thrombus formation was observed in any of the retrieved pumps. A modified Model II with a new connector system was subsequently intra-thoracically implanted in a sheep. The sheep survived for 482 days without any sign of thromboembolic complication or hemolysis at a fixed rotational speed of 1700 rpm and an average pump flow rate of 5 L/min. There was no intra-device thrombus formation or infarct in major organs. The Model III system, consisting of an implantable controller and a new MSCP with a reduced input power of 13 W, has been developed and implanted in a chronic sheep model. The MSCP was implanted in the left pleural space and the controller in the abdominal wall. The experiment is still in progress for more than 30 days without any significant complication to date. These animal studies strongly suggest the feasibility of the MSCP for use as long-term circulatory assist. PMID- 10667730 TI - Development of an antithrombogenic and antitraumatic blood pump: the Gyro C1E3. AB - The Gyro C1E3 is a centrifugal blood pump. Its antithrombogenic and antitraumatic blood features were demonstrated by prior studies. Based upon these studies, a mass production model of the C1E3 is becoming commercially available. Therefore, this feasibility study was conducted using the mass production models of the Gyro C1E3 for long-term cardiac assist in ex vivo animal experiments. Five healthy calves were used and 15 pump heads were applied for different time periods (Group 1, 30 days; Group 2, 14 days; Group 3, 10 and 7 days; Group 4, 4 days; and Group 5, 2 days). Activated clotting time (ACT) was kept at 200-250 sec. All five calves demonstrated neither abnormal signs nor abnormal blood examination data throughout the experiment. During necropsy, no thromboembolism was found in any downstream organs. Groups 1-4 showed thrombi inside the pump heads while two pumps in Group 5 had no thrombi formations. Bearing deformation or possible wear did not increase after 2 days of pumping. The C1E3 is capable of long-term assist circulation. However, after 2 days of pumping, careful observation is necessary since thrombi may occur inside the pump when ACT is controlled under 250 sec. During the weaning stage or low flow (under 2 L/min), over 250 sec of ACT is recommended to assure the safety of the patient. PMID- 10667731 TI - Clinical evaluation of the Gyro Pump C1E3 as a cardiopulmonary bypass pump. AB - The Gyro Pump C1E3 is a new centrifugal pump with numerous features, including a ceramic pivot bearing system, secondary vanes, and an eccentric inlet port. To evaluate its biocompatibility, antithrombogenicity, and produced hemolysis, we used the Gyro Pump during cardiopulmonary bypass (CPB) for coronary artery bypass grafting (CABG) cases to compare it with the BioMedicus pump. From September 1998 to February 1999, 30 consecutive patients underwent CABG under conventional CPB. Fifteen patients were supported by the Gyro Pump C1E3 (Group G), and the remaining 15 patients, by a BioMedicus BP-80 pump (Group B). In both groups, flow rate was equivalent. Blood samples were taken as follows: preoperative, 60 minutes after the end of the procedure, and at postoperative days (POD) 0, 1, and 2. We evaluated the plasma free hemoglobin (free Hb) as an indication of hemolysis; beta-thromboglobulin (beta-TG) and platelet factor four (PF-4) as an indication of platelet deterioration; C3, C4, CH50 for complement activation; coagulation parameters, fibrinolytic factor, thrombomodulin, nitric oxide (NO), and endothelin as an indication of endothelial deterioration. This was the first clinical sized Gyro Pump CIE3. De-airing from the pump was easily accomplished via the eccentric oblique inlet port. The system, including its console, was easily and simply controlled. Perioperative laboratory data were not markedly changed in either group with demonstrated equivalence for biocompatibility and hemolysis. After pumping, no thrombus formation or pivot wear were observed inside the pump. This atraumatic, small centrifugal pump appears well suited not only for CPB but also for circulatory support. PMID- 10667732 TI - Application of a new dynamic flow model for investigating the biocompatibility of modified surfaces. AB - An in vitro model was developed to compare the biocompatibility of four different coating methods (three heparin and one nonheparin) under hemodynamic conditions. Fresh human donor blood (heparin 5 IU/ml) was recirculated in a standardized experimental circuit. All circuit components were either coated or remained uncoated for control purposes. The aim of the study was to investigate a wide spectrum of effects on blood; coagulation parameters (e.g., fibrinogen, ATIII, thrombin-antithrombin-complex), complement parameters (C1rsC1 Inh, C3b(Bb)P, SC5b 9, C5a), differential blood analyses, platelet activation (flow cytometric investigations), PF 4, and PMN-elastase release were examined by showing possible trends. All heparin coated systems reduced platelet stimulation in comparison to untreated biomaterials. Leukocyte activation was reduced to different degrees depending upon the coating method used. Complement activation was markedly reduced by all coated systems. The results obtained indicate that the pump driven, dynamic blood flow model is suitable to characterize the biocompatibility of surface modified biomaterials. Advantages lie in the integration of the different polymers as parts of the circuit, the low priming volume, and the generation of blood flow conditions similar to those that occur in clinical applications. PMID- 10667733 TI - Abnormal electrical stimulus of an intra-aortic balloon pump with concurrent support with continuous veno-venous hemodialysis. AB - Malfunction of electronic medical support apparatus utilized in the ICU usually causes system failure. We report several occurrences of a potentially dangerous interaction between a continuous veno-venous hemodialysis (CVVHD) system and an intra-aortic balloon pump (IABP) counterpulsation device in four patients requiring both systems. The patients had acute renal failure in the face of multi organ failure and were dependent upon the balloon pump for pressure support. Electrical interference created by the roller pump action of the CVVHD system was identified by the balloon pump as cardiac in origin, and it responded by inflation and deflation. As the blood pump rate was reduced, the interference reduced to the point of complete cessation when the blood pump was shut down. Whereas one patient transiently had a significant drop of mean arterial pressure (from 70 +/- 4 to 40 +/- 2 mm Hg) the other observed occurrences had no clinically significant sequelae. Electrocardiogram (ECG) tracings identified the abnormal stimulus and systematic review identified as potential sources for the creation of this interference static electricity buildup, piezoelectric properties of the polyvinyl chloride tubing, and, possibly but less likely, radiofrequency interference. A newer generation ECG cable and advanced cardiac rhythm recognition software (CardioSync) have been introduced with the Datascope System 98, and the ECG interference, although still occasionally present, does not cause erratic inflation and deflation of the intra-aortic balloon pump. Interference between different electrical support systems may occur, and we suggest that the systems be tested for compatibility before combined use and that older equipment be more rigorously tested for potential clinically significant interference. PMID- 10667734 TI - Life-threatening acute systemic lupus erythematosus: survival after multiple extracorporeal modalities: a place for the multipotential extracorporeal service. AB - Diffuse alveolar hemorrhage secondary to systemic lupus erythematosus (SLE) may cause life-threatening respiratory failure and may be associated with multiple organ failure. Extensive support may be necessary to sustain life while systemic therapy becomes effective. We report here a patient with profound respiratory failure secondary to SLE associated with multiorgan failure, who was supported with veno-arterial extracorporeal lung assist (ECLA), veno-venous ECLA, and multiple continuous renal replacement therapies during plasmapheresis. The full spectrum of extracorporeal life support and treatment modalities was performed seamlessly by a single service within the critical care department. PMID- 10667735 TI - Physiology of nonpulsatile circulation: acute versus chronic support. PMID- 10667736 TI - At issue: schizophrenia and rheumatoid arthritis: the negative association revisited. AB - A strong negative association between schizophrenia and rheumatoid arthritis (RA), implying low comorbidity, has been found in 12 of 14 previous studies, which we review. To this literature we add two recently acquired data sets encompassing 28,953 schizophrenia patients, only 31 of whom had comorbid RA. Integrating our new data into those of the previous nine studies, which stratified their populations according to psychiatric diagnosis, we obtain a median frequency of RA in schizophrenia populations of 0.09 percent and a mean frequency of 0.66 percent, well below the expected range of 1 percent. These data robustly support prior studies. We also present a meta-analysis evaluating the association between the two diseases by integrating information derived from nine data sets, each furnishing an estimate of the relative risk of RA in schizophrenia patients versus that in other psychiatric patients. We find that the estimated rate of RA among schizophrenia patients is only 29 percent of the corresponding rate in other psychiatric patients. Further, the relative risk of RA in schizophrenia patients versus that in the general population is even less than 29 percent and could be as low as one-third of this value. We present a new hypothesis involving the platelet activating factor system in an effort to account for this negative association and review the suggestions of other investigators toward this end. Finally, we consider the glutamatergic system dysfunction hypothesis of schizophrenia and suggest a possible common pharmacological approach that may ameliorate some of the symptomatology of both schizophrenia and RA. PMID- 10667737 TI - Schizophrenia: an epigenetic puzzle? AB - Developments in molecular biology over the past three decades have led to an increasing awareness of the importance of epigenetic phenomena in a variety of genome functions. Epigenetic aspects of complex multifactorial diseases including schizophrenia, however, have not been investigated sufficiently. Various facets of epigenetics are reevaluated through their putative relevance to four theories of schizophrenia: neurodevelopmental, dopamine dysfunction, viral, and genetic anticipation with unstable DNA. The heuristic value of the epigenetic model of schizophrenia arises from the possibility of integration of a wide variety of empirical data into a new theoretical framework. It can be hypothesized that in addition to pathological effects of DNA structural mutations and environmental factors, inherited and acquired epigenetic defects, or epimutations, may be of etiological importance in schizophrenia. In addition, the epigenetic model may lead to experiments investigating the molecular substrates of genetic environmental interactions. PMID- 10667738 TI - Effects of cognitive treatment in psychiatric rehabilitation. AB - Ninety subjects with severe and disabling psychiatric conditions, predominantly schizophrenia, participated in a controlled-outcome trial of the cognitive component of Integrated Psychological Therapy (IPT), a group-therapy modality intended to reestablish basic neurocognitive functions. The cognitive therapy was delivered to subjects in the experimental condition during intensive 6-month treatment periods. Control subjects received supportive group therapy. Before, during, and after the intensive treatment period, all subjects received an enriched regimen of comprehensive psychiatric rehabilitation, including social and living skills training, optimal pharmacotherapy, occupational therapy, and milieu-based behavioral treatment. IPT subjects showed incrementally greater gains compared with controls on the primary outcome measure, the Assessment of Interpersonal Problem-Solving Skills, suggesting that procedures that target cognitive impairments of schizophrenia spectrum disorders can enhance patients' response to standard psychiatric rehabilitation, at least in the short term, in the domain of social competence. There was equivocal evidence for greater improvement in the experimental condition on the Brief Psychiatric Rating Scale disorganization factor and strong evidence for greater improvement on a laboratory measure of attentional processing. There was significant improvement in both conditions on measures of attention, memory, and executive functioning, providing support for the hypothesis that therapeutic procedures that target impaired cognition enhance response to conventional psychiatric rehabilitation modalities over a 6-month timeframe. PMID- 10667739 TI - Developmental theory for a cognitive enhancement therapy of schizophrenia. AB - Recent findings on psychosocial and neurodevelopmental anomalies in schizophrenia patients indicate that deficits related to social cognition-the ability to act wisely in social interactions-may be important constraints on complete social and vocational recovery. Social cognition is acquired over many decades and appears to be partially independent of formal IQ and neuropsychological problems. It invites a more developmental approach to the rehabilitation of schizophrenia, one that we call Cognitive Enhancement Therapy (CET). CET draws on an emerging literature that implicates both pre- and postonset neurodevelopmental difficulties, as well as a complementary literature on diffuse neuropsychological impairments that supports the notion of a neurodevelopmental insult. We analyzed evidence for an associated developmental basis to social cognitive impairment in the context of a model that addressed both the acquisition of interpersonal wisdom and the adaptive process that might follow developmental failures. A contemporary model of human cognition is then used to identify the metacognitive functions that characterize the developmental acquisition of normal cognition and, by inference, the associated difficulties of many patients with schizophrenia. A rehabilitation strategy for schizophrenia, designed to facilitate the metacognitive transition from prepubertal to young adult social cognition, would thus emphasize developmental learning experiences during the remediation of social cognitive deficits. A "gistful" appraisal of interpersonal behavior and novel social contexts best reflects the theoretical intent of this new intervention. PMID- 10667740 TI - Practice principles of cognitive enhancement therapy for schizophrenia. AB - Cognitive Enhancement Therapy (CET) is a developmental approach to the rehabilitation of schizophrenia patients that attempts to facilitate an abstracting and "gistful" social cognition as a compensatory alternative to the more demanding and controlled cognitive strategies that often characterize schizophrenia as well as much of its treatment. Selected cognitive processes that developmentally underlie the capacity to acquire adult social cognition have been operationalized in the form of relevant interactive software and social group exercises. Treatment methods address the impairments, disabilities, and social handicaps associated with cognitive styles that appear to underlie the positive, negative, and disorganized symptom domains of schizophrenia. Style-related failures in secondary rather than primary socialization, particularly social cognitive deficits in context appraisal and perspective taking, are targeted goals. Illustrative examples of the techniques used to address social and nonsocial cognitive deficits are provided, together with encouraging preliminary observations regarding the efficacy of CET. PMID- 10667741 TI - How long to wait for a response to clozapine: a comparison of time course of response to clozapine and conventional antipsychotic medication in refractory schizophrenia. AB - This study compared the time course to clinical improvement with clozapine and with conventional antipsychotic medications. A double-blind trial compared clozapine and haloperidol in patients with schizophrenia who were refractory to conventional antipsychotic medication and were hospitalized for 30 to 364 days at 15 Veteran Affairs medical centers during the year before study entry. Patients in the original study were randomly assigned to haloperidol or clozapine and followed for 12 months, at maximum tolerable doses. Patients who completed a full year of treatment with clozapine (n = 122), or with either haloperidol or another conventional antipsychotic medication (n = 123) and who also completed the 9- or 12-month assessment were included. Response to treatment was defined as 20 percent improvement on standard scales of symptoms and quality of life at the latter of the 9- or 12-month interviews. More patients assigned to clozapine achieved 20 percent improvement in symptoms at each followup. Among patients who did not improve at 6 weeks, 3 months, or 6 months, there were no significant differences between clozapine and comparison patients in outcomes at 1 year. Among patients who did improve, maintenance of that improvement also did not differ between the groups at 1 year on symptom measures. Maintenance of improvement in quality of life at 1 year was significantly greater for clozapine patients who had improved at 6 months (p < 0.04). Significant differential symptom response to clozapine occurred exclusively during the first 6 weeks of treatment. PMID- 10667742 TI - Risperidone in the treatment of first-episode psychotic patients: a double-blind multicenter study. Risperidone Working Group. AB - An international, multicenter, double-blind study was conducted in 183 patients with a first psychotic episode (provisional schizophreniform disorder or schizophrenia; DSM-III-R) treated with flexible doses of risperidone or haloperidol for 6 weeks. At endpoint, 63 percent of risperidone-treated patients and 56 percent of haloperidol-treated patients were clinically improved (> or = 50% reduction in Positive and Negative Syndrome Scale total scores). Risperidone was better tolerated than haloperidol: the severity of extrapyramidal symptoms was significantly lower in the risperidone-treated patients; significantly fewer risperidone-treated patients required antiparkinsonian medication; and significantly fewer discontinued treatment because of adverse events. A post hoc analysis revealed that low doses of these antipsychotics were efficacious in some patients. Furthermore, the severity of extrapyramidal symptoms and the use of antiparkinsonian medications were significantly lower in patients receiving low doses (maximum, < or = 6 mg/day) than high doses (maximum, > 6 mg/day) of risperidone or haloperidol. These findings are consistent with the suggestion that patients with a first psychotic episode may require low doses of antipsychotic medications. Studies designed specifically to compare low and high doses of antipsychotics are warranted to help optimize treatment for these patients. PMID- 10667743 TI - Tardive dyskinesia: possible involvement of free radicals and treatment with vitamin E. AB - A decade ago a hypothesis introduced to explain tardive dyskinesia (TD) implicated free radicals generated secondary to neuroleptic treatment. Since then many preclinical and clinical studies have investigated this possibility. These studies suggest that free radicals are probably involved in the pathogenesis of TD and that vitamin E could be efficacious in its treatment. PMID- 10667744 TI - Tardive dystonia. AB - This paper provides an overview of the phenomenology, epidemiology, and treatment of tardive dystonia. Tardive dystonia is one of the extrapyramidal syndromes that starts after long-term use of dopamine receptor antagonists. The diagnosis is based on the presence of chronic dystonia, defined as a syndrome of sustained muscle contractions, frequently causing twisting and repetitive movements or abnormal postures. Furthermore, dystonia must develop either during or within 3 months of a course of antipsychotic treatment, and other causes such as Wilson's disease, acute dystonia, or a conversion reaction must be ruled out. Tardive dystonia occurs in about 3 percent of patients on long-term antipsychotic treatment. Some probable risk factors for tardive dystonia are younger age, male, and the presence of tardive dyskinesia. The treatment of tardive dystonia starts with an evaluation of the need for using the causative drug. If antipsychotics must be continued, a switch to an atypical antipsychotic, particularly clozapine, may be helpful. If the dystonia is relatively localized, botulinum toxin is an effective but not well-known treatment possibility. If tardive dystonia is more extensive, either dopamine-depleting drugs or high dosages of anticholinergics can be tried. PMID- 10667745 TI - The "benefits" of distractibility: mechanisms underlying increased Stroop effects in schizophrenia. AB - Recent studies of selective attention in schizophrenia patients suggest a particular pattern of single-trial Stroop performance: increased facilitation but not interference in reaction times (RTs), combined with increased error interference. Our Stroop task analysis suggests that this pattern can be explained by a selective attention deficit if one accounts for (1) performance in the congruent condition; (2) the nature of the neutral stimulus; (3) the relationship between accuracy and RT; and (4) response set effects. To test these hypotheses, we examined Stroop performance in 40 DSM-IV schizophrenia patients and 20 healthy control subjects, using a range of neutral stimuli (color patches, noncolor words, color words not in the response set). The findings confirmed several of our predictions and the results were consistent with the hypothesis that abnormal Stroop performance in schizophrenia reflects a failure to adequately attend to the task-appropriate stimulus dimension (color). This inattention affects both the congruent and incongruent conditions and multiple points in the information processing pathway. PMID- 10667747 TI - Sensory gating in rats: lack of correlation between auditory evoked potential gating and prepulse inhibition. AB - This study was designed to evaluate the possible similarities between two paradigms designed to measure sensory gating: (1) an auditory evoked potential (AEP), called the P50 gating paradigm; and (2) an acoustic startle (ASR), called the prepulse inhibition paradigm. These paradigms show a number of methodological, pharmacological, and neurobiological similarities, and they are both disturbed in patients with schizophrenia. In the first of three experiments, the AEP gating and the ASR gating were measured in rats. Although both AEP and ASR gating could readily be obtained, there appeared to be no correlation between the performance in these two paradigms. This lack of correlation was confirmed using a factor analytical approach, where the AEP gating and the ASR gating parameters were found to load on different factors. In the second experiment, the interstimulus interval in the ASR paradigm was increased to 500 ms (identical to the interstimulus interval of the AEP gating paradigm). This increase reduced the degree of ASR gating, although some gating could still be obtained. Again no correlation was found between AEP and ASR gating, and this was again confirmed by the factorial analysis. In the final experiment, the effects of the dopamine D2/3 agonist 7-OHDPAT were evaluated in both paradigms. This selective agonist dose dependently reduced ASR gating but had no effect on AEP gating. Together, these data strongly suggest that AEP and ASR gating measure two different aspects of information processing and indicate that both paradigms may be important for investigating the neurobiological disturbances observed in patients with psychoses. PMID- 10667746 TI - Panmodal processing imprecision as a basis for dysfunction of transient memory storage systems in schizophrenia. AB - Schizophrenia is a severe mental disorder associated with cognitive disturbances that may reflect underlying deficits in the functioning of brain transient memory storage systems. This study investigates performance in three distinct tasks that require transient memory storage: (1) tone discrimination, (2) object weight discrimination, and (3) "AX"-type visual continuous performance task. The tests used were chosen to investigate the degree to which a similar pattern of performance deficit could be observed across multiple sensory and cognitive domains in schizophrenia. In each of the paradigms, a similar pattern emerged: subjects with schizophrenia showed severe performance deficits whenever performance depended on functioning of transient memory systems. The deficits were apparent at both short and long interstimulus intervals (ISI), however, and schizophrenia subjects were no more affected by increasing ISI than were controls. Moreover, when short ISI performance was matched across groups by manipulating task difficulty, subsequent decay in performance was equivalent across groups. Thus, although schizophrenia subjects show severe performance deficits in memory-dependent tasks, the deficits do not appear to reflect impaired transient memory per se. Rather, they appear to reflect impaired precision of operation of such systems, irrespective of the duration over which representations must be maintained. The severe deficits in processing precision, despite the relatively preserved maintenance of representation, may be relevant to pathophysiological models of schizophrenia. PMID- 10667748 TI - An artificial neural network that uses eye-tracking performance to identify patients with schizophrenia. AB - Several researchers have underscored the importance of precise characterization of eye-tracking dysfunction (ETD) in patients with schizophrenia. This biological trait appears to be useful in estimating the probability of genetic recombination in an individual, so it may be helpful in linkage studies. This article describes a nonlinear computational model for using ETD to identify schizophrenia. A back propagation neural network (BPNN) was used to classify schizophrenia patients and normal control subjects on the basis of their eye-tracking performance. Better classification results were obtained with BPNN than with a linear computational model (discriminant analysis): a priori predictions were approximately 80 percent correct. These results suggest, first, that eye-tracking patterns can be useful in distinguishing patients with schizophrenia from a normal comparison group with an accuracy of approximately 80 percent. Second, parallel distributed processing networks are able to detect higher order nonlinear relationships among predictor quantitative measurements of eye-tracking performance. PMID- 10667749 TI - Premorbid personality in psychoses. AB - This study aimed to establish correlates of the dimensions of schizophrenia in the premorbid personality traits of patients. A sample of 112 patients of relatively recent illness onset who were admitted for a psychotic episode were assessed with a semistructured interview for schizophrenia. Positive and negative symptoms were evaluated with the Scale for the Assessment of Positive Symptoms and the Scale for the Assessment of Negative Symptoms at the time of hospital discharge; positive, negative, and disorganization scores were obtained from these scales. Premorbid personality was assessed blindly through a partially modified version of the Personality Assessment Schedule using interviews with the parents or a close relative. Schizoid traits were significantly associated with negative and positive dimensions. Sociopathic traits were related to the disorganization dimension. Trends toward significance were obtained between passive-dependent traits and the negative and disorganization dimensions, and between the schizotypal dimension and the positive dimension. Partial correlational analyses were used to control for the effect of the remaining personality dimensions on the above relationships. Schizoid premorbid traits were still significantly related to the negative dimension but to a lesser degree to the disorganization and positive dimensions. The association between sociopathic premorbid traits and the disorganization dimension remained significant. These results suggest the preexistence of a three-dimensional structure predisposing to psychoses within the premorbid personality; this structure is more evident in patients with short illness duration. PMID- 10667750 TI - Nonresponding schizophrenia: differentiation by neurological soft signs and neuropsychological tests. AB - Schizophrenia patients have higher scores on neurological soft-signs (NSS) and show greater deficits on a variety of neuropsychological tests than normal control subjects and mixed groups of psychiatric patients. Among chronic schizophrenia patients it is unclear which of these types of deficits most strongly differentiates patients who remain consistently symptomatic in spite of treatment with several conventional neuroleptics (nonresponders) as compared with relapsing chronic schizophrenia patients who improve substantially with treatment (relative responders). In this study, 25 nonresponders and 20 relative responders to conventional neuroleptics were compared on an NSS battery and a limited number of neuropsychological tests, which evaluated deficits influenced by functioning of frontal and nonfrontal brain areas. NSS scores showed the largest difference between relative responders and nonresponders, and statistical analyses suggested that NSS scores were the strongest differentiator between the two groups of chronic schizophrenia patients. Scores differentiating the two groups involved deficits influenced by both frontal and nonfrontal functioning. A predominance of negative symptoms in the current clinical picture was highly correlated with high NSS scores. PMID- 10667751 TI - Familial liability to schizophrenia: a sibling study of negative symptoms. AB - Negative symptoms are important features in schizophrenia, so in milder form they might also serve as indicators of "unexpressed" liability to schizophrenia among patients' adult relatives without schizophrenia. To address this question, we assessed negative symptoms in 39 stable schizophrenia or schizoaffective outpatients, 39 of their siblings, 38 well control probands, and 38 of their siblings. Negative symptom measures included standard behavior ratings of the core negative symptoms of affective flattening and alogia, as well as a self report measure of social anhedonia. As expected, even stable outpatients with schizophrenia exhibited significantly more negative symptoms than control probands and control siblings. However, negative behavioral symptoms of affective flattening, alogia, and anhedonia did not significantly differentiate the siblings of the schizophrenia patients from the control probands or their siblings, although there were some trends for anhedonia. The findings suggest that core negative symptoms of observed affective flattening and poverty of speech are not likely to be useful as strong indicators of "unexpressed" liability to schizophrenia. PMID- 10667752 TI - Dimensions of psychosis in affected sibling pairs. AB - Factor analytical studies of schizophrenia symptoms have consistently suggested three or more symptom dimensions, but it is not known whether any of these dimensions have a genetic basis. The purpose of this study was to investigate to what extent the dimensions show familial aggregation. Symptom ratings were made using the SAPS and SANS and the OPCRIT checklist on the members of 109 sibling pairs with DSM-IV schizophrenia or schizoaffective disorder. Factor analyses were performed on the ratings of both instruments, and correlations were made of within-pair factor scores. Analyses were also performed on the 89 pairs in which both members had a diagnosis of schizophrenia. Factor analysis of SAPS and SANS ratings resulted in positive, negative, and disorganization factors; analysis of OPCRIT ratings resulted in positive, negative, disorganization, and first-rank delusion factors. Only the disorganization dimension showed significant within pair correlations, but these were of modest size and not significantly greater than the correlations for the other dimensions. None of the dimensions showed sufficient familial aggregation to suggest that they are close markers of genetic or common environmental factors that contribute liability to schizophrenia. They may be weakly associated with such factors and with factors that do not contribute liability to schizophrenia but do influence the form taken by the illness. PMID- 10667753 TI - Communication disturbances in relatives beyond the age of risk for schizophrenia and their associations with symptoms in patients. AB - This article provides a detailed examination of subclinical disturbances in the natural speech of healthy relatives beyond the age of risk for schizophrenia. Speech samples from 43 stable schizophrenia outpatients, 42 nonschizophrenia parents of patients (pairs only), and 23 control subjects matched to the parents were analyzed for frequencies of six specific types of communication failures. The parents had higher overall communication disturbance ratings than the control subjects. The specific types of failures that occurred more frequently were unclarities caused by (1) language structural breakdown, (2) use of vague, overinclusive words, and (3) use of words with ambiguous meanings. In intrafamilial analyses, higher levels of communication disturbance in parents were associated with greater severity of illness in their patient offspring. These results support the idea that communication disturbances may be one manifestation of a stable genetic vulnerability to schizophrenia. The nature of the failures identified suggests the possible involvement of weaknesses in specific areas of cognitive functioning. PMID- 10667754 TI - First person account: schizophrenia, substance abuse, and HIV. PMID- 10667755 TI - Can biological phenomena be understood by humans? PMID- 10667756 TI - All parties keen to press on with Europe-based science website. PMID- 10667757 TI - Clinton proposes $2.8 billion increase in science funding. PMID- 10667758 TI - Yale hopes a $500 million boost will raise research profile. PMID- 10667759 TI - NIH cancer researchers to get free access to 'OncoMouse'. PMID- 10667760 TI - Ministries cooperate to plan Japanese genome centre. PMID- 10667761 TI - Issue of patents on 'Dolly' technology stirs controversy. PMID- 10667762 TI - US survey reveals location of human tissue samples. PMID- 10667763 TI - Head of US watchdog faces uncertain future. PMID- 10667764 TI - Top UK epidemiologist suspended after complaints. PMID- 10667765 TI - Cell contamination leads to inaccurate data: we must take action now. PMID- 10667766 TI - Mitchell saw the new vista, if not the details. PMID- 10667767 TI - Science moves to centre stage. PMID- 10667768 TI - New approaches to old age. PMID- 10667769 TI - Pluto story. PMID- 10667770 TI - Nogo in nerve regeneration. PMID- 10667771 TI - Palaeoclimatology. Out of Africa. PMID- 10667772 TI - Harvesting sunlight safely. PMID- 10667773 TI - One photon seen by one electron PMID- 10667774 TI - Rice, microbes and methane. PMID- 10667775 TI - Fluid mechanics. Jets from a singular surface PMID- 10667777 TI - Volcanic action at Axial Seamount PMID- 10667776 TI - Evolutionary psychology meets g. PMID- 10667778 TI - Energy for microbial life on Europa. PMID- 10667779 TI - A function for guttural pouches in the horse. PMID- 10667780 TI - Inhibitor of neurite outgrowth in humans. PMID- 10667781 TI - Creating the narrowest carbon nanotubes. PMID- 10667782 TI - Nodal signalling in vertebrate development. AB - Communication between cells during early embryogenesis establishes the basic organization of the vertebrate body plan. Recent work suggests that a signalling pathway centering on Nodal, a transforming growth factor beta-related signal, is responsible for many of the events that configure the vertebrate embryo. The activity of Nodal signals is regulated extracellularly by EGF-CFC cofactors and antagonists of the Lefty and Cerberus families of proteins, allowing precise control of mesoderm and endoderm formation, the positioning of the anterior posterior axis, neural patterning and left-right axis specification. PMID- 10667783 TI - A pigment-binding protein essential for regulation of photosynthetic light harvesting. AB - Photosynthetic light harvesting in plants is regulated in response to changes in incident light intensity. Absorption of light that exceeds a plant's capacity for fixation of CO2 results in thermal dissipation of excitation energy in the pigment antenna of photosystem II by a poorly understood mechanism. This regulatory process, termed nonphotochemical quenching, maintains the balance between dissipation and utilization of light energy to minimize generation of oxidizing molecules, thereby protecting the plant against photo-oxidative damage. To identify specific proteins that are involved in nonphotochemical quenching, we have isolated mutants of Arabidopsis thaliana that cannot dissipate excess absorbed light energy. Here we show that the gene encoding PsbS, an intrinsic chlorophyll-binding protein of photosystem II, is necessary for nonphotochemical quenching but not for efficient light harvesting and photosynthesis. These results indicate that PsbS may be the site for nonphotochemical quenching, a finding that has implications for the functional evolution of pigment-binding proteins. PMID- 10667784 TI - Probing bulk states of correlated electron systems by high-resolution resonance photoemission AB - Electron correlations are known to play an important role in determining the unusual physical properties of a variety of compounds. Such properties include high-temperature superconductivity, heavy fermion behaviour and metal-to insulator transitions. High-resolution photoelectron spectroscopy (PES) provides a means of directly probing the electronic states (particularly those near the Fermi level) in these materials, but the short photoelectron mean free paths (< or = 5 A) associated with the low excitation energies conventionally used (< or = 120 eV) make this a surface-sensitive technique. Now that high-resolution PES is possible at much higher energies, with mean free paths as long as 15 A (ref. 6), it should become feasible to probe the bulk electronic states in these materials. Here we demonstrate the power of this technique by applying it to the cerium compounds CeRu2Si2 and CeRu2. Previous PES studies of these compounds revealed very similar spectra for the Ce 4f electronic states, yet it is expected that such states should be different owing to their differing degrees of hybridization with other valence bands. Our determination of the bulk Ce 4f electronic states of these compounds resolves these differences. PMID- 10667785 TI - Dynamic instabilities and memory effects in vortex matter AB - The magnetic flux line lattice in type II superconductors serves as a useful system in which to study condensed matter flow, as its dynamic properties are tunable. Recent studies have shown a number of puzzling phenomena associated with vortex motion, including: low-frequency noise and slow voltage oscillations; a history-dependent dynamic response, and memory of the direction, amplitude duration and frequency of the previously applied current; high vortex mobility for alternating current, but no apparent vortex motion for direct currents; and strong suppression of an a.c. response by small d.c. bias. Taken together, these phenomena are incompatible with current understanding of vortex dynamics. Here we report a generic mechanism that accounts for these observations. Our model, which is derived from investigations of the current distribution across single crystals of NbSe2, is based on a competition between the injection of a disordered vortex phase at the sample edges, and the dynamic annealing of this metastable disorder by the transport current. For an alternating current, only narrow regions near the edges are in the disordered phase, while for d.c. bias, most of the sample is in the disordered phase--preventing vortex motion because of more efficient pinning. The resulting spatial dependence of the disordered vortex system serves as an active memory of the previous history. PMID- 10667786 TI - Singularity dynamics in curvature collapse and jet eruption on a fluid surface AB - Finite-time singularities--local divergences in the amplitude or gradient of a physical observable at a particular time--occur in a diverse range of physical systems. Examples include singularities capable of damaging optical fibres and lasers in nonlinear optical systems, and gravitational singularities associated with black holes. In fluid systems, the formation of finite-time singularities cause spray and air-bubble entrainment, processes which influence air-sea interaction on a global scale. Singularities driven by surface tension have been studied in the break-up of pendant drops and liquid sheets. Here we report a theoretical and experimental study of the generation of a singularity by inertial focusing, in which no break-up of the fluid surface occurs. Inertial forces cause a collapse of the surface that leads to jet formation; our analysis, which includes surface tension effects, predicts that the surface profiles should be describable by a single universal exponent. These theoretical predictions correlate closely with our experimental measurements of a collapsing surface singularity. The solution can be generalized to apply to a broad class of singular phenomena. PMID- 10667787 TI - A single-photon detector in the far-infrared range AB - The far-infrared region (wavelengths in the range 10 microm-1 mm) is one of the richest areas of spectroscopic research, encompassing the rotational spectra of molecules and vibrational spectra of solids, liquids and gases. But studies in this spectral region are hampered by the absence of sensitive detectors--despite recent efforts to improve superconducting bolometers, attainable sensitivities are currently far below the level of single-photon detection. This is in marked contrast to the visible and near-infrared regions (wavelengths shorter than about 1.5 microm), in which single-photon counting is possible using photomultiplier tubes. Here we report the detection of single far-infrared photons in the wavelength range 175-210 microm (6.0-7.1 meV), using a single-electron transistor consisting of a semiconductor quantum dot in high magnetic field. We detect, with a time resolution of a millisecond, an incident flux of 0.1 photons per second on an effective detector area of 0.1 mm2--a sensitivity that exceeds previously reported values by a factor of more than 10(4). The sensitivity is a consequence of the unconventional detection mechanism, in which one absorbed photon leads to a current of 10(6)-10(12) electrons through the quantum dot. By contrast, mechanisms of conventional detectors or photon assisted tunnelling in single electron transistors produce only a few electrons per incident photon. PMID- 10667788 TI - Efficient organic photovoltaic diodes based on doped pentacene. AB - Recent work on solar cells based on interpenetrating polymer networks and solid state dye-sensitized devices shows that efficient solar-energy conversion is possible using organic materials. Further, it has been demonstrated that the performance of photovoltaic devices based on small molecules can be effectively enhanced by doping the organic material with electron-accepting molecules. But as inorganic solar cells show much higher efficiencies, well above 15 per cent, the practical utility of organic-based cells will require their fabrication by lower cost techniques, ideally on flexible substrates. Here we demonstrate efficiency enhancement by molecular doping in Schottky-type photovoltaic diodes based on pentacene--an organic semiconductor that has received much attention as a promising material for organic thin-film transistors, but relatively little attention for use in photovoltaic devices. The incorporation of the dopant improves the internal quantum efficiency by more than five orders of magnitude and yields an external energy conversion efficiency as high as 2.4 per cent for a standard solar spectrum. Thin-film devices based on doped pentacene therefore appear promising for the production of efficient 'plastic' solar cells. PMID- 10667789 TI - Rainfall and drought in equatorial east Africa during the past 1,100 years. AB - Knowledge of natural long-term rainfall variability is essential for water resource and land-use management in sub-humid regions of the world. In tropical Africa, data relevant to determining this variability are scarce because of the lack of long instrumental climate records and the limited potential of standard high-resolution proxy records such as tree rings and ice cores. Here we present a decade-scale reconstruction of rainfall and drought in equatorial east Africa over the past 1,100 years, based on lake-level and salinity fluctuations of Lake Naivasha (Kenya) inferred from three different palaeolimnological proxies: sediment stratigraphy and the species compositions of fossil diatom and midge assemblages. Our data indicate that, over the past millennium, equatorial east Africa has alternated between contrasting climate conditions, with significantly drier climate than today during the 'Medieval Warm Period' (approximately AD 1000 1270) and a relatively wet climate during the 'Little Ice Age' (approximately AD 1270-1850) which was interrupted by three prolonged dry episodes. We also find strong chronological links between the reconstructed history of natural long-term rainfall variation and the pre-colonial cultural history of east Africa, highlighting the importance of a detailed knowledge of natural long-term rainfall fluctuations for sustainable socio-economic development. PMID- 10667790 TI - Precise climate monitoring using complementary satellite data sets AB - Observations from Earth-orbiting satellites have been a key component in monitoring climate change for the past two decades. This has become possible with the availability of air temperatures from the Microwave Sounding Unit (MSU) since 1979, sea surface temperatures from the Advanced Very High Resolution Radiometer (AVHRR) since 1982 and, most recently, measurements of atmospheric water vapour content from the Special Sensor Microwave Imager (SSM/I) since 1987. Here we present a detailed comparison of each pair of these three time series, focusing on both interannual and decadal variations in climate. We find a strong association between sea surface temperature, lower-tropospheric air temperature and total column water-vapour content over large oceanic regions on both time scales. This lends observational support to the idea of a constant relative humidity model having a moist adiabatic lapse rate. On the decadal timescale, the combination of data sets shows a consistent warming and moistening trend of the marine atmosphere for 1987-1998. PMID- 10667791 TI - The Pleistocene serpent Wonambi and the early evolution of snakes. AB - The Madtsoiidae were medium sized to gigantic snakes with a fossil record extending from the mid-Cretaceous to the Pleistocene, and spanning Europe, Africa, Madagascar, South America and Australia. This widely distributed group survived for about 90 million years (70% of known ophidian history), and potentially provides important insights into the origin and early evolution of snakes. However, madtsoiids are known mostly from their vertebrae, and their skull morphology and phylogenetic affinities have been enigmatic. Here we report new Australian material of Wonambi, one of the last-surviving madtsoiids, that allows the first detailed assessment of madtsoiid cranial anatomy and relationships. Despite its recent age, which could have overlapped with human history in Australia, Wonambi is one of the most primitive snakes known--as basal as the Cretaceous forms Pachyrhachis and Dinilysia. None of these three primitive snake lineages shows features associated with burrowing, nor do any of the nearest lizard relatives of snakes (varanoids). These phylogenetic conclusions contradict the widely held 'subterranean' theory of snake origins, and instead imply that burrowing snakes (scolecophidians and anilioids) acquired their fossorial adaptations after the evolution of the snake body form and jaw apparatus in a large aquatic or (surface-active) terrestrial ancestor. PMID- 10667792 TI - Stimulation by ammonium-based fertilizers of methane oxidation in soil around rice roots. AB - Methane is involved in a number of chemical and physical processes in the Earth's atmosphere, including global warming. Atmospheric methane originates mainly from biogenic sources, such as rice paddies and natural wetlands; the former account for at least 30% of the global annual emission of methane to the atmosphere. As an increase of rice production by 60% is the most appropriate way to sustain the estimated increase of the human population during the next three decades, intensified global fertilizer application will be necessary: but it is known that an increase of the commonly used ammonium-based fertilizers can enhance methane emission from rice agriculture. Approximately 10-30% of the methane produced by methanogens in rice paddies is consumed by methane-oxidizing bacteria associated with the roots of rice; these bacteria are generally thought to be inhibited by ammonium-based fertilizers, as was demonstrated for soils and sediments. In contrast, we show here that the activity and growth of such bacteria in the root zone of rice plants are stimulated after fertilization. Using a combination of radioactive fingerprinting and molecular biology techniques, we identify the bacteria responsible for this effect. We expect that our results will make necessary a re-evaluation of the link between fertilizer use and methane emissions, with effects on global warming studies. PMID- 10667793 TI - Activin- and Nodal-related factors control antero-posterior patterning of the zebrafish embryo. AB - Definition of cell fates along the dorso-ventral axis depends on an antagonistic relationship between ventralizing transforming growth factor-beta superfamily members, the bone morphogenetic proteins and factors secreted from the dorsal organizer, such as Noggin and Chordin. The extracellular binding of the last group to the bone morphogenetic proteins prevents them from activating their receptors, and the relative ventralizer:antagonist ratio is thought to specify different dorso-ventral cell fates. Here, by taking advantage of a non-genetic interference method using a specific competitive inhibitor, the Lefty-related gene product Antivin, we provide evidence that cell fate along the antero posterior axis of the zebrafish embryo is controlled by the morphogenetic activity of another transforming growth factor-beta superfamily subgroup--the Activin and Nodal-related factors. Increasing antivin doses progressively deleted posterior fates within the ectoderm, eventually resulting in the removal of all fates except forebrain and eyes. In contrast, overexpression of activin or nodal related factors converted ectoderm that was fated to be forebrain into more posterior ectodermal or mesendodermal fates. We propose that modulation of intercellular signalling by Antivin/Activin and Nodal-related factors provides a mechanism for the graded establishment of cell fates along the antero-posterior axis of the zebrafish embryo. PMID- 10667794 TI - Separable processing of consonants and vowels. AB - There are two views about the nature of consonants and vowels. One view holds that they are categorically distinct objects that play a fundamental role in the construction of syllables in speech production. The other view is that they are convenient labels for distinguishing between peak (vowel) and non-peak (consonant) parts of a continuous stream of sound that varies in sonority (roughly the degree of openness of the vocal apparatus during speech), or that they are summary labels for bundles of feature segments. Taking the latter view, consonants and vowels do not have an independent status in language processing. Here we provide evidence for the possible categorical distinction between consonants and vowels in the brain. We report the performance of two Italian speaking aphasics who show contrasting, selective difficulties in producing vowels and consonants. Their performance in producing individual consonants is independent of the sonority value and feature properties of the consonants. This pattern of results suggests that consonants and vowels are processed by distinct neural mechanisms, thereby providing evidence for their independent status in language production. PMID- 10667795 TI - Noradrenaline in the ventral forebrain is critical for opiate withdrawal-induced aversion. AB - Cessation of drug use in chronic opiate abusers produces a severe withdrawal syndrome that is highly aversive, and avoidance of withdrawal or associated stimuli is a major factor contributing to opiate abuse. Increased noradrenaline in the brain has long been implicated in opiate withdrawal, but it has not been clear which noradrenergic systems are involved. Here we show that microinjection of beta-noradrenergic-receptor antagonists, or of an alpha2-receptor agonist, into the bed nucleus of the stria terminalis (BNST) in rats markedly attenuates opiate-withdrawal-induced conditioned place aversion. Immunohistochemical studies revealed that numerous BNST-projecting cells in the A1 and A2 noradrenergic cell groups of the caudal medulla were activated during withdrawal. Lesion of these ascending medullary projections also greatly reduced opiate-withdrawal-induced place aversion, whereas lesion of locus coeruleus noradrenergic projections had no effect on opiate-withdrawal behaviour. We conclude that noradrenergic inputs to the BNST from the caudal medulla are critically involved in the aversiveness of opiate withdrawal. PMID- 10667796 TI - Nogo-A is a myelin-associated neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-1. AB - The capacity of the adult brain and spinal cord to repair lesions by axonal regeneration or compensatory fibre growth is extremely limited. A monoclonal antibody (IN-1) raised against NI-220/250, a myelin protein that is a potent inhibitor of neurite growth, promoted axonal regeneration and compensatory plasticity following lesions of the central nervous system (CNS) in adult rats. Here we report the cloning of nogo A, the rat complementary DNA encoding NI 220/250. The nogo gene encodes at least three major protein products (Nogo-A, -B and -C). Recombinant Nogo-A is recognized by monoclonal antibody IN-1, and it inhibits neurite outgrowth from dorsal root ganglia and spreading of 3T3 fibroblasts in an IN-1-sensitive manner. Antibodies against Nogo-A stain CNS myelin and oligodendrocytes and allow dorsal root ganglion neurites to grow on CNS myelin and into optic nerve explants. These data show that Nogo-A is a potent inhibitor of neurite growth and an IN-1 antigen produced by oligodendrocytes, and may allow the generation of new reagents to enhance CNS regeneration and plasticity. PMID- 10667797 TI - Identification of the Nogo inhibitor of axon regeneration as a Reticulon protein. AB - Adult mammalian axon regeneration is generally successful in the peripheral nervous system (PNS) but is dismally poor in the central nervous system (CNS). However, many classes of CNS axons can extend for long distances in peripheral nerve grafts. A comparison of myelin from the CNS and the PNS has revealed that CNS white matter is selectively inhibitory for axonal outgrowth. Several components of CNS white matter, NI35, NI250(Nogo) and MAG, that have inhibitory activity for axon extension have been described. The IN-1 antibody, which recognizes NI35 and NI250(Nogo), allows moderate degrees of axonal regeneration and functional recovery after spinal cord injury. Here we identify Nogo as a member of the Reticulon family, Reticulon 4-A. Nogo is expressed by oligodendrocytes but not by Schwann cells, and associates primarily with the endoplasmic reticulum. A 66-residue lumenal/extracellular domain inhibits axonal extension and collapses dorsal root ganglion growth cones. In contrast to Nogo, Reticulon 1 and 3 are not expressed by oligodendrocytes, and the 66-residue lumenal/extracellular domains from Reticulon 1, 2 and 3 do not inhibit axonal regeneration. These data provide a molecular basis to assess the contribution of Nogo to the failure of axonal regeneration in the adult CNS. PMID- 10667798 TI - Torque-generating units of the flagellar motor of Escherichia coli have a high duty ratio. AB - Rotation of the bacterial flagellar motor is driven by an ensemble of torque generating units containing the proteins MotA and MotB. Here, by inducing expression of MotA in motA- cells under conditions of low viscous load, we show that the limiting speed of the motor is independent of the number of units: at vanishing load, one unit turns the motor as rapidly as many. This result indicates that each unit may remain attached to the rotor for most of its mechanochemical cycle, that is, that it has a high duty ratio. Thus, torque generators behave more like kinesin, the protein that moves vesicles along microtubules, than myosin, the protein that powers muscle. However, their translation rates, stepping frequencies and power outputs are much higher, being greater than 30 microm s(-1), 12 kHz and 1.5 x 10(5) pN nm s(-1), respectively. PMID- 10667799 TI - The DExH protein NPH-II is a processive and directional motor for unwinding RNA. AB - All aspects of cellular RNA metabolism and processing involve DExH/D proteins, which are a family of enzymes that unwind or manipulate RNA in an ATP-dependent fashion. DExH/D proteins are also essential for the replication of many viruses, and therefore provide targets for the development of therapeutics. All DExH/D proteins characterized to date hydrolyse nucleoside triphosphates and, in most cases, this activity is stimulated by the addition of RNA or DNA. Several members of the family unwind RNA duplexes in an NTP-dependent fashion in vitro; therefore it has been proposed that DExH/D proteins couple NTP hydrolysis to RNA conformational change in complex macromolecular assemblies. Despite the central role of DExH/D proteins, their mechanism of RNA helicase activity remains unknown. Here we show that the DExH protein NPH-II unwinds RNA duplexes in a processive, unidirectional fashion with a step size of roughly one-half helix turn. We show that there is a quantitative connection between ATP utilization and helicase processivity, thereby providing direct evidence that DExH/D proteins can function as molecular motors on RNA. PMID- 10667800 TI - DNA-bound structures and mutants reveal abasic DNA binding by APE1 and DNA repair coordination [corrected]. AB - Non-coding apurinic/apyrimidinic (AP) sites in DNA are continually created in cells both spontaneously and by damage-specific DNA glycosylases. The biologically critical human base excision repair enzyme APE1 cleaves the DNA sugar-phosphate backbone at a position 5' of AP sites to prime DNA repair synthesis. Here we report three co-crystal structures of human APE1 bound to abasic DNA which show that APE1 uses a rigid, pre-formed, positively charged surface to kink the DNA helix and engulf the AP-DNA strand. APE1 inserts loops into both the DNA major and minor grooves and binds a flipped-out AP site in a pocket that excludes DNA bases and racemized beta-anomer AP sites. Both the APE1 active-site geometry and a complex with cleaved AP-DNA and Mn2+ support a testable structure-based catalytic mechanism. Alanine substitutions of the residues that penetrate the DNA helix unexpectedly show that human APE1 is structurally optimized to retain the cleaved DNA product. These structural and mutational results show how APE1 probably displaces bound glycosylases and retains the nicked DNA product, suggesting that APE1 acts in vivo to coordinate the orderly transfer of unstable DNA damage intermediates between the excision and synthesis steps of DNA repair. PMID- 10667801 TI - Design of single-layer beta-sheets without a hydrophobic core. AB - The hydrophobic effect is the main thermodynamic driving force in the folding of water-soluble proteins. Exclusion of nonpolar moieties from aqueous solvent results in the formation of a hydrophobic core in a protein, which has been generally considered essential for specifying and stabilizing the folded structures of proteins. Outer surface protein A (OspA) from Borrelia burgdorferi contains a three-stranded beta-sheet segment which connects two globular domains. Although this single-layer beta-sheet segment is exposed to solvent on both faces and thus does not contain a hydrophobic core, the segment has a high conformational stability. Here we report the engineering of OspA variants that contain larger single-layer beta-sheets (comprising five and seven beta-strands) by duplicating a beta-hairpin unit within the beta-sheet. Nuclear magnetic resonance and small-angle X-ray scattering analyses reveal that these extended single-layer beta-sheets are formed as designed, and amide hydrogen-deuterium exchange and chemical denaturation show that they are stable. Thus, interactions within the beta-hairpin unit and those between adjacent units, which do not involve the formation of a hydrophobic core, are sufficient to specify and stabilize the single-layer beta-sheet structure. Our results provide an expanded view of protein folding, misfolding and design. PMID- 10667802 TI - Bioengineering programmes rise to meet the challenge of a young science. PMID- 10667803 TI - Exploring the territory in tissue engineering. PMID- 10667804 TI - Progress from a fragile start. PMID- 10667805 TI - The immunobiology of transplantation. AB - Having recently published A History of Transplantation Immunology, I have become acutely aware of the great revolution that has taken place in the field of organ transplantation - not only in terms of our understanding of the immunobiological mechanisms underlying the rejection process and the various means of prolonging graft survival, but also in clinical terms. Only 50 years ago, clinical kidney allo-transplantation (and it was the kidney that spearheaded clinical advances) was a highly experimental and dangerous procedure dependent on whole-body irradiation and the results were often intensely disappointing. Now, the transplantation of kidneys is almost routine, results are excellent and the lessons learnt about immunosuppression and patient management have been applied with great success to a large number of other organs (see below). PMID- 10667806 TI - Immunobiology of solid organ transplantation. AB - The field of organ transplantation is entering a very exciting phase in which tolerance induction may become a therapeutic possibility. The induction of tolerance would allow patients to enjoy the benefits of their transplanted organ without risking the mortality and morbidity associated with long-term pharmacological immunosuppression. In this review, we explore the immunobiology of solid organ transplantation, discussing the immunological mechanisms responsible for allograft rejection, and outlining the rational behind a range of successful experimental tolerance induction strategies. PMID- 10667807 TI - Renal transplantation: origins and future. AB - Renal transplantation from its first successful operation in 1954 has become the preferred treatment option of patients with renal failure. The success of this recent development is followed in this article which tabulates surgical, tissue typing and immunosuppressive advancement. This success has produced other problems notably chronic rejection, posttransplant lympho-proliferative disease and donor shortage. The solutions to these problems are discussed, together with a hint of what is to come in the future. PMID- 10667808 TI - Present status and future prospects in liver transplantation. AB - Results of liver transplantation (LTx) have largely improved over the last few years and a one year patient survival rate of around 90% for elective transplants can now be reached. This is mostly the consequence of better patient selection and preparation, improved surgical techniques, improved intra- and postoperative care, reduced incidence of primary non function (< or =5%) and hepatic artery thrombosis (< or =5%), the understanding that the liver induces a tolerogenic response and the development of more judicious immunosuppressive protocols. Because patient and graft survival have dramatically improved, other problems become more overt and need to be addressed: side effects of long-term exposure to immunosuppression, recurrent disease, development of biliary strictures. LTx for acute liver failure continues to bring special difficulties. Strategies need to be urgently developed to augment the number of donor grafts and to combat the profound organ shortage - currently the only limiting factor to wider application of LTx. PMID- 10667809 TI - Pancreatic transplantation: a review. AB - Pancreatic transplantation recently became a routine treatment for Type I diabetic patients with uremia or for those who previously received a kidney transplant with 1 year graft and patient survival of over 80% and 90%, respectively. Despite the life-long need for immunosuppression, this is clearly acceptable when compared to the need for dialysis and insulin therapy, and it reduces the evolution of diabetic complications. Isolated pancreatic transplant is less commonly applied because of the need for immunosuppression and the high rate of complications. However, this can still be an acceptable option for individual patients with brittle diabetes and hypoglycemic unawareness. Despite the fact that pancreas transplantation is an effective treatment for selected Type I diabetics, it remains a difficult surgical procedure with many potential complications and with several issues still subject to debate. In this article, the authors describe the procedure in all of its aspects and variations, and offer, through a review of the recent literature, insights on the current status of this transplant PMID- 10667810 TI - Small bowel transplantation. AB - Many patients die each year lacking only a functional small bowel to survive. The minimum amount of small intestinal absorptive surface required to sustain life varies from patient to patient. Prolonged survival with oral alimentation alone has been reported in a few patients with an intact duodenum and as little as 15 45 cm of residual jejunum. However, without long-term total parenteral nutrition (TPN), prolonged patient survival is the exception rather than the rule. Chronic parenteral nutrition is associated with complications, including sepsis, venous thrombosis, metabolic disorders and liver dysfunction. From studies of patients currently on long-term TPN, it would appear that there are between two and three patients per million of population per year who develop irreversible small bowel failure. It is estimated that 20 new patients/year in the UK receiving home TPN would be potential candidates for small bowel transplantation. PMID- 10667811 TI - Surgical procedures for primary, metastatic or adjacent parotid tumours. AB - OBJECTIVE: The retrospective analysis of the surgical procedures in primary parotid and metastatic or adjacent parotid tumors. PATIENTS AND METHODS: Retrospective review of the records of 145 patients operated on for primary, metastatic or adjacent parotid tumors revealed 85 patients with benign tumors, 24 with primary malignant tumors, 19 with squamous skin carcinomas, 12 with skin melanomas, 3 with basocellular carcinomas and 2 with sarcomas of the parotid region. The analysis included the type of parotidectomy, the need for facial nerve sacrifice (FNS), type of neck dissection and soft part reconstruction. RESULTS: Superficial parotidectomy was performed in 81% of the benign parotid tumors and 100% of skin melanomas. Total parotidectomy was frequent in malignant parotid tumors (62%), epidermoid skin tumors (64%) and in basocellular/sarcomas of the parotid region (80%). Skin graft or flaps was infrequent in primary malignant tumors (12.5%), and frequent in epidermoid skin tumors (74%), melanomas (58%) and basocellular/sarcomas (100%). FNS was necessary in primary malignant (25%), adjacent epidermoid (37%), melanomas (17%) and basocellular/sarcomas (80%). Details on neck dissections are provided. CONCLUSIONS: Superficial parotidectomy was an adequate procedure for most benign parotid tumors and for melanoma patients. In primary malignant and adjacent or metastatic skin tumors, total parotidectomy, neck dissection and soft part reconstruction were frequent procedures. FNS and soft part reconstruction should be anticipated more frequently in squamous/basocellular skin tumors or sarcomas adjacent to the parotid gland. PMID- 10667812 TI - Surgical repair of pectus carinatum. AB - Pectus carinatum represents a variety of protrusion deformities of the anterior chest wall. Although various non-operative methods of treatment have been employed, surgery has been widely accepted as the only effective method for the correction of pectus carinatum. We evaluate our 14 year single center experience of pectus carinatum correction on 111 patients using a uniform technique of internal stabilization employing stainless steel struts. Operative correction required double bilateral chondrotomy parasternally and at points of transition to normal ribs, followed by detorsion of the sternum, retrosternal mobilization and correction of the everted sternum as well as of the everted and inverted ribs. The mobilized sternum after incomplete wedge osteotomy was finally stabilized by one transternal and two bilateral parasternal metal struts. The corrections were completed with successful repair in 109 patients (98.2%). Major recurrences in 2 patients (1.8%) were corrected while mild recurrence were observed in 3 patients (2.7%). PMID- 10667813 TI - Significant prognostic factors in patients with node-negative gastric cancer. AB - BACKGROUND: We evaluated the influence of several clinicopathological variables on 5 year survival of patients with node-negative gastric cancer. PATIENTS AND METHODS: Clinical characteristics were retrieved from the records of all patients who underwent gastric resection between 1985 and 1995 at the Department of General Surgery, Sendai National Hospital, and follow-up data were obtained from our tumor registry. Pathological characteristics were determined from a detailed review of all available histopathological slides. The results of a retrospective analysis of clinicopathological data of 339 patients having no lymph node metastasis were compared with those of 358 patients with lymph node metastasis. Univariate and multivariate analyses of patients with node-negative gastric cancer were performed to evaluate the prognostic significance of clinicopathological features (age, gender, gross type, histological type, depth of invasion and location). RESULTS: The 5 year survival rate for patients with node-negative gastric cancer was 92.5%. Node-negative gastric cancers were characterized by a smaller tumor, expansive and medullary histological type, and less frequency of lymphatic invasion and vascular permeation. In multivariate analysis, the statistical significant prognostic factors were tumor size (P = 0.0185), vascular permeation (P = 0.0011) and cancer-stromal relationship (P = 0.0291). CONCLUSION: Tumor size, vascular microinvasion and cancer-stromal relationship are the most reliable predictors of 5 year survival for patients with node-negative gastric cancer. PMID- 10667814 TI - Study of survival and prognostic factors in patients undergoing resection for gastric linitis plastica: a review of 86 cases. AB - A retrospective study was conducted in a series of 86 patients (51 men and 35 women; mean age 63.4 years) treated from 1979 to 1995 for linitis plastica of the stomach (LP). The mean interval between the first manifestations and surgery was 3.5 months. The most frequent clinical sign was epigastric pain which occurred in 80.4% of cases. Biopsies were positive in 75.6% of cases. Typical features of LP were found in only 46% of esogastric barium enemas and 11.8% of upper gastrofiberscopic examinations. Seventy-four patients had surgical excision (51 total and 23 partial gastrectomies). There were 6 (7%) postoperative deaths and 10 (11.6%) surgical complications. Node involvement was found in 54 (72.9%) patients. Overall actuarial survival (n = 86) was 50% at 12 months, 40% at 18 months and 7.5% at 84 months. Survival did not depend on the delay in diagnosis, histological analysis of the extremities of the excised piece, associated tissue differentiation, node involvement or the type of surgical excision. The prognosis differed according to tumor height (P<0.01) and involvement of the deep stomach wall (P<0.001). No independent prognostic factor was found in multivariate analysis. Surgery remains the sole possibility for curative therapy in these patients. PMID- 10667815 TI - Gallbladder carcinoma during laparoscopic cholecystectomy: is it associated with bad prognosis? AB - Laparoscopic cholecystectomy is the treatment of choice for gallstone disease. The ultrasonogram has failed for the early detection of gallbladder cancer, especially if inflammation (chronic or acute) is present. Incidental gallbladder could be an important cancer finding during laparoscopic cholecystectomy, due to the potential cancer cell dissemination during the procedure. In our Department, 6500 laparoscopic cholecystectomies have been performed in the last 5 years and in 15 cases (0.23%) gallbladder cancer was found during surgery or after histological examination of the resected gallbladder. In none of these 15 patients was pre-operative diagnosis of gallbladder carcinoma postulated. When re evaluation of the pre-operative ultrasonograms was done, it was possible to observe signs suggesting the presence of neoplastic infiltration in 4 of them (28.6%). During videoscopic exploration, also in 4 patients, the suspicion of gallbladder cancer was noted. Laparoscopic cholecystectomy was completed in 9 patients. In 2 of them, in situ or mucosal invasion was demonstrated with a long survival. One patient presented recurrence at the biliary hilum 2,5 years after surgery. Six patients were re-operated and in 4 of them peritoneal or port site metastasis was found; all died early (4.5 month median survival). The other 2 patients were submitted to liver bed resection and lymph node dissection. These patients are free of cancer recurrence after 15 months of follow-up. Six patients were converted to open surgery, performing palliative procedures and died before the 12 month follow-up. The suspicion of pre-operative gallbladder cancer is generally unlikely to be confirmed based on ultrasonographic signs; but, in some cases with high suspicion, further investigation (TAC, tumor markers, etc.) must be indicated in order to avoid poor results. Laparoscopic cholecystectomy could be associated with bad prognosis, and then, when gallbladder cancer is suspected during the laparoscopic procedure, conversion to open surgery could be the best choice. PMID- 10667816 TI - Prevention of postherniorrhaphy persistent pain: results of a prospective study. AB - Anterior tension-free and laparoscopic inguinal herniorrhaphies represent one of the most common surgical procedure. Postherniorrhaphy persistent pain due to injures of inguinal regional nerves is rare, difficult to cure, often disabling and involving malpractice litigation. In a prospective study, we evaluated the effectiveness of neurectomy of the iliohypogastric nerve in prevention of postoperative persistent pain after anterior tension free herniorrhaphy. Between 1992-1995, we performed 180 anterior herniorraphies in 151 male patients. Iliohypogastric nerve was removed in all the herniorrhaphies. Polypropylene plug and sutured mesh were employed. Postoperative pain and clinical relevance of hypo anesthesia and paresthesia were assessed. No patient complained of postoperative persistent pain. Hypo-anesthesia, never considered incapacitating, was present in 1% of patients after 2 years. We consider neurectomy of the iliohypogastric nerve a potentially useful surgical step in preventing postoperative persistent pain after anterior tension-free herniorrhaphy. PMID- 10667817 TI - Invasive and non-invasive physiological monitoring of blunt trauma patients in the early period after emergency admission. AB - Pulmonary artery catheterization is usually not available to critically injured patients before admission to the intensive care unit, where action to correct values derived from such monitoring may be too late. Methods allowing hemodynamic monitoring during the early stages after trauma need to be explored. We used non invasive monitoring systems (bioimpedance cardiac output monitoring, pulse oximetry and transcutaneous oximetry) to evaluate early temporal hemodynamic patterns after blunt trauma, and compared these to invasive PA monitoring. We included prospectively 134 patients monitored shortly after admission to the emergency department. The non-invasive impedance cardiac output estimations under extenuating emergency conditions approximated those of the thermodilution method: r = 0.83, r2 = 0.69, P<0.001; bias and precision were -0.02+/-0.78 l/min/m2. In the intensive care unit, these values improved further to: r = 0.91, r2 = 0.83, P<0.001; bias and precision = 0.36+/-0.59 l/min/m2. Monitoring revealed episodes of hypotension, low cardiac index, arterial hemoglobin desaturation, low transcutaneous oxygen and high transcutaneous carbon dioxide tensions, and low oxygen consumption during initial resuscitation. Low flow and poor tissue perfusion were more pronounced in non-survivors by both methods. Multicomponent non-invasive monitoring systems give continuous on-line, real-time displays of physiological data that allow early recognition of circulatory dysfunction. Such systems provide information similar to that provided by the invasive thermodilution method, and are easier and safer to use. PMID- 10667818 TI - Intrapericardial tumbling bullet. AB - Foreign bodies of the pericardium are rare and they are associated most commonly with significant trauma. The diagnosis of a pericardial foreign body can be difficult. One must distinguish between foreign matter in the cardiac chamber or free-floating in the mediastinum. Serial chest X-rays and fluoroscopy were most helpful to us. Neither CT scan nor an echocardiogram were particularly helpful. To prevent pericarditis, either sterile or non-sterile, with potential for other significant complications, removal of a pericardial foreign body is always indicated. PMID- 10667819 TI - Histopathological findings in experimental aneurysms embolized with conventional and thrombogenic/antithrombolytic Guglielmi coils. AB - We studied the short- and long-term histological responses induced by conventional and modified electronically detachable coils (GDCs) in experimental aneurysms. Eighteen carotid bifurcation aneurysms were produced microsurgically in rabbits. Six animals each were treated either with conventional or with GDCs coated with a mixture of tissue-thromboplastin to enhance intra-aneurysmal thrombus formation and of plasminogen activator inhibitor type-1 (PAI-1) in inhibit intra-aneurysmal clot fibrinolysis. Six served as untreated controls. Follow-up angiograms were obtained immediately and at 3, 6, 9, 12, 17, and 24 weeks after embolization prior to sacrifice of the animals. All aneurysms were studied macroscopically and histopathologically with the coils in situ. Five of six control aneurysms remained patent. Endovascular occlusion rates between > 90% and 100% were achieved in nine of twelve coiled aneurysms. Follow-up angiography demonstrated recanalization and coil compaction in 5 of them. Gross and microscopic histopathological examination revealed a membrane covering the orifice, intra-aneurysmal scar formation, and development of a neo-intima in both treatment groups at 17 and 24 weeks postembolization. The granulation tissue response appeared to be equally distributed in aneurysms treated with either uncoated or coated coils. Further quantitative morphometric studies are needed to prove if a thrombogenic/antithrombolytic coil-coating might be of value in providing a more enduring anatomic result after GDC-treatment of human brain aneurysms. PMID- 10667820 TI - Postoperative headache after the lateral suboccipital approach: craniotomy versus craniectomy. AB - The lateral suboccipital approach to the cerebellopontine angle is typically performed as a small craniectomy. Incisional pain and headache following cerebellopontine angle surgery have been reported. Adherence of the cervical muscles to the dura, which is richly innervated, with consequent traction has been suggested to be responsible for postoperative headache. Therefore, postoperative headache probably could be reduced by replacing the bone flap between the muscles and the dura. In a prospective non-randomized study this hypothesis was tested by comparing craniectomy and craniotomy. 40 patients underwent removal of an acoustic neuroma via the retrosigmoid approach. Patients with a history of migraine, with additional intracerebral tumors or recurrencies as well as patients who developed a CSF fistula postoperatively were excluded. 29 patients were eligible for further evaluation. 13 patients underwent a craniotomy, 16 patients a craniectomy. All patients were subject to a standardized telephone interview three months and one year after surgery. Comparing the craniotomy group to the craniectomy group no difference was observed regarding age, sex, tumor size and duration of operation. 3 months as well as 12 months postoperatively headache was significantly (p < 0.05) less frequent in the craniotomy group as compared to the craniectomy group. In conclusion, an osteoplastic craniotomy significantly reduces postoperative headache and is therefore highly recommended. PMID- 10667821 TI - A new technique for attaching a stereotactic frame to the head: technical note. AB - The aim of this study was to develop a device which allows an intermediate, painless fixation of a stereotactic frame prior to definite pin fixation. To stabilize the stereotactic frame rubber coated metal springs were used. By testing the springs on 30 volunteers with different head diameters and circumferences the optimal shape was determined. In the clinical setting 15 patients undergoing stereotactic surgery were tested, stability and patient's convenience were measured. The procedure was well tolerated and measurements revealed symmetric distances between head and frame. Therefore these metal springs are a useful accessory to the Leksell stereotactic system. PMID- 10667822 TI - Surgical management for supratentorial astrocytic tumors. AB - To compare the surgical treatment of supratentorial astrocytic tumors, various methods were performed by the same surgeon. Removal of the tumor was performed using stereotactic open surgery, the fluorescein surgical microscope, and a frameless stereotactic system, and these methods were compared. The method using the stereotactic technique was useful because there was no disturbance by the shifting of the brain during the operation. However, its limitation was that only points can be marked. The fluorescein surgical microscope was very useful in the cases where neuroradiological images were enhanced by the contrast medium, but deep lesions could not be identified from the brain surface. This method could not be used, either, in the case of images that were not enhanced. By the method using the frameless stereotactic system, identification of tumors including deep lesions was possible from every direction, but the problems were the mobility of the registered skin and the shifting of the brain during the operation. On the basis of these results, the combined method of the fluorescein surgical microscope and the frameless stereotactic system appeared to be useful when neuroradiological images of lesions were enhanced because these methods were complementary towards each other, and the frameless stereotactic system supplemented by the stereotactic open surgery technique (such as leaving a marker in deep lesions just before the start of microsurgery) seemed useful when images could not be enhanced. PMID- 10667823 TI - Frameless neuronavigation applied to endoscopic neurosurgery. AB - OBJECTIVE: We retrospectively analyzed the indications, surgical techniques, and applicability of frameless neuronavigation to endoscopic procedures in a heterogeneous group of 15 patients. MATERIAL AND METHODS: In 8 patients indications for surgery were cystic lesions, in 3 patients intraventricular tumors, and in 4 patients occlusive hydrocephalus. The mean age was 39 years (range 9-76 years). The follow-up period ranged from 5-24 months (mean 10 months). Frameless neuronavigation was accomplished with the "operating arm system" in 10 cases and with the "optical tracking system" in 5 cases (RADIONICS, Burlington, USA). RESULTS: In all 15 cases, neuronavigation sufficiently provided anatomical orientation, preoperative planning, and intraoperative realization of the approach. The calculated mean calibration error was 2.1 mm. There have been no permanent morbidities and no mortalities related to the use of endoscopes and neuronavigation. CONCLUSION: In endoscopic neurosurgery, frameless neuronavigation is a useful tool in planning and realizing the approach and improving intraoperative orientation in selected cases. Indications are small or hidden lesions, impaired visual conditions, abnormal anatomy, and narrow ventricles. Endoscopic procedures include fenestration and resection of intraventricular or intraparenchymal cysts, biopsy of intraventricular tumors, and third ventriculostomy in selected cases. PMID- 10667824 TI - Optimized visualization of contrast medium dependent differences in CT/MRI scans by RGB transformation. AB - OBJECTIVE: In order to be able to assess the contrast medium enhancement of vessels and regions with disturbed blood-brain barrier, the neurosurgeon must mentally perform a subtraction of the corresponding native and contrast-enhanced scans of the CT or MRI. The principle disadvantages of this comparison are the amount of time required and the potential errors which may result from false interpretation. METHODS: The process presented here combines corresponding scans without and with contrast medium offline on a PC. In this process each couple of the black and white scans is assigned to the colour channels of the RGB system (red, green, blue) and transformed to one colour picture of the RGB colour space (additive colour mixture). RESULTS: We demonstrate that contrast medium-dependent changes can be represented in a colour contrast of complementary colours in one single picture. Additionally, the unchanged structures remain in the black and white contrast as they were before. CONCLUSION: Thanks to the optimized visualization of contrast medium dependent differences there are besides saving in time further advantages such as the reduced amount of pictures and an objective representation. PMID- 10667825 TI - Endoscopic third ventriculostomy: a study of intracranial pressure vs. haemodynamic changes. AB - Fourteen paediatric patients with obstructive hydrocephalus were studied. They underwent endoscopic third ventriculostomy under general anaesthesia. Their ages ranged from 1 to 144 weeks (mean 34+/-36 weeks) and weight from 2 to 22 kg (mean 10.2+/-5.4 kg). In an attempt to identify the possible mechanisms of the intraoperative haemodynamic changes associated with endoscopic third ventriculostomy, we studied the intracranial pressure measured in the third ventricle versus the haemodynamic changes. The intracranial pressure was measured using a pressure transducer attached at one end to the endoscope and the other end to the monitor. The mean third ventricle pressure value was 10.2 mmHg (+/ 3.5). Bradycardia occurred in six (43%) of our patients. The mean value of the lowest heart rate reading intraoperatively was 81 beats/min (+/-31.8). Negative correlation was obtained between the intracranial pressure and the haemodynamic changes. Alerting the surgeon to perforate the floor of the third ventricle or withdraw the scope away from it was sufficient to resolve the bradycardia. We concluded that serious dysrhythmias might occur during endoscopic third ventriculostomy, the majority of which can be resolved without medications. PMID- 10667826 TI - The tanycytic ependymoma of the lateral ventricle: case report. AB - The tanycytic ependymoma is an extremely rare, primitive neuroectodermal tumor, arising from the ependymoglial cells or tanycytes. Such cells are generally seen in the primitive nervous system instead of the mature ependymal cells. The tanycytic ependymoma described in this report was found in a 42-year-old man. Histological analysis strongly suggested that this tumor originated from a primitive progenitor cell, the ependymoglia or the tanycyte in the lateral ventricle. PMID- 10667827 TI - Pituitary abscess secondary to isolated sphenoid sinusitis. AB - Intracranial complications from isolated sphenoid sinusitis are rare but nevertheless demonstrate both a high morbidity and mortality. We herein report a case of a pituitary abscess secondary to sphenoid sinusitis in a 12-year-old boy. This patient presented with an acute onset of moderate fever and headache, followed by progressive right ptosis. An emergency endoscopic endonasal sphenoidotomy with sinus drainage and postoperative antibiotic therapy resulted in a satisfactory recovery. PMID- 10667828 TI - Orbital foreign bodies after penetrating gunshot wounds: retrospective analysis of 22 cases and clinical review. AB - We conducted a retrospective analysis of 22 patients having orbital penetrating gunshot wounds treated over a 4-years period. The neurological status and the site of injury for each patient are evaluated in this study. We propose a practical protocol in the management of these orbital foreign bodies. Surgical treatment was performed in 4 patients (had functional deficit) with medial orbitotomy in 2, lateral orbitotomy in 1, and superior orbitotomy in 1. 3 of them are improved, in one case the blindness has been continued. 18 patients were treated conservatively and all of them are improved. All patients were followed up for 2 years with cranial X-rays and CT scans. Neurological sequelae were regressed which existed before the surgery. In conservatively treated cases, infection, migration and functional deficit were not seen. In conclusion, orbital penetrating gunshot wounds must be evaluated precisely by the surgeon and this evaluation sets the guidelines for management. The operation should be reserved for the patients in whom the necrotic soft tissues or orbital damages restrict ocular movements. PMID- 10667829 TI - Comparison of a new automatically controlled electrocoagulator (Valleylab NS 2000 INSTANT RESPONSE technology) with a high-frequency coagulator. AB - Bipolar electrocoagulation is one of the most important procedures in modern neurosurgery. However, there are still many practical problems, especially tissue adherence to the tips of the coagulating forceps and the difficulty removing carbonized clots from the tips. Both make the process less accurate and more time consuming. To prevent formation of coagulum, recently, irrigation with a saline solution and coating of the forceps tips with a special metal have been tried. In this work, we compare a new bipolar electrocoagulator with automatic output control in relation to tissue impedance (Auto Suture - Valleylab NS 2000 with INSTANT RESPONSE technology) with a high-frequency coagulator (Erbotom ICC 350, Erbe). The femoral arteries and nerves of Wistar rats, weighing on average 360 g, were prepared and coagulation was carried out with variable power settings during a constant time (3 seconds). Sections were stained with haematoxylin-eosin, van Gieson and Luxol-Fast-Blue for histological examination. Coagulation with Erbotom ICC 350 resulted in tissue sticking to the tips of the forceps in all cases, regardless of the power chosen. With the new electrocoagulator, tissue adherence to the forceps tips was not seen. With the new system, effective coagulation was also achieved at comparably lower power settings. PMID- 10667830 TI - Problems and perspectives of phenotyping for drug-metabolizing enzymes in man. AB - Pronounced interindividual differences in drug disposition are mainly caused by differences in the activity of liver drug-metabolizing enzymes. These depend on known and unknown covariates, including genetic as well as environmental factors. Phenotyping, i.e. assessment of enzyme activities in vivo after administration of a test dose, seems to be a promising tool for determining actual metabolic capacities. Although it is a well-established experimental approach, phenotyping has not yet found its way into clinical practice. Main reasons for this are lack of validation for many probes and assays used, complicated procedures, invasiveness, semi-quantitative test results, non-compliance on behalf of the subjects tested, high costs, and lack of prospective clinical studies to assess the benefit of phenotyping for patients. Problems and perspectives of phenotyping are exemplified for the cytochrome P-450 enzymes CYP1A2 and CYP3A4, two major human drug-metabolizing enzymes. PMID- 10667831 TI - Decreased oral bioavailability of loxoprofen at second administration in human subjects. AB - The objective of this study was to determine the extent of period effect on the pharmacokineitcs of loxoprofen during consecutive dosing. Loxipen and Loxonin tablets were administered to 16 healthy Korean male subjects at a single dose of 60 mg as loxoprofen sodium anhydrous in a 2 x 2 crossover investigation with a two-week wash-out phase. Concentrations of loxoprofen in plasma were measured by HPLC method for 6 h. The two formulations were found bioequivalent, but analysis of variance (ANOVA) indicated that there was a significant (p < 0.05) period effect in AUCinf (area under the plasma concentration-time curve from time zero to infinity) between the administrations. A 20% decrease in the AUC was seen at the second administration. This period effect on pharmacokinetics of loxoprofen may be relevant for the patients who need consecutive administration of the drug. PMID- 10667832 TI - Evaluation of the efficacy and dose-response relationship of dexibuprofen (S(+) ibuprofen) in patients with osteoarthritis of the hip and comparison with racemic ibuprofen using the WOMAC osteoarthritis index. AB - OBJECTIVE: Treatment with non-steroidal anti-inflammatory drugs is the most common pharmacological therapy of rheumatic diseases. For the symptomatic treatment of painful disorders a dose-response relationship of the NSAID should be a basic requirement, which is difficult to be proven in studies because rheumatic diseases are heterogenous in terms of clinical involvement. The aim of this double-blind randomized trial was to compare the isolated active enantiomer dexibuprofen (S(+)-ibuprofen) with the double dose of racemic ibuprofen and to show a dose-response relationship of dexibuprofen in painful osteoarthritis of the hip. METHODS: 178 patients were randomly assigned to dexibuprofen 600/1,200 mg or racemic ibuprofen 2,400 mg daily. The primary endpoint was the improvement of the WOMAC osteoarthritis index after 15 days of therapy. The analysis was by intention to treat. RESULTS: The evaluation of the WOMAC OA index showed statistically significant equivalence of dexibuprofen 400 mg t.i.d. compared with racemic ibuprofen 800 mg t.i.d. by a Mann-Whitney statistic of 0.578 and the corresponding lower bound of the 95% confidence interval of 0.498. The test for superiority of dexibuprofen was borderline significant with p = 0.055. Dexibuprofen 400 mg t.i.d. and dexibuprofen 200 mg t.i.d. showed a statistically significant dose-response relationship in improving the WOMAC OA index (p = 0.023). Patients suffered from adverse drug reactions, mainly gastrointestinal disorders, 13.34% on dexibuprofen 200 mg, 15.25% on dexibuprofen 400 mg and 16.94% on racemic ibuprofen 800 mg. CONCLUSIONS: The active enantiomer dexibuprofen (S(+)-ibuprofen) proved to be an effective non-steroidal anti inflammatory drug with a significant dose-response relationship in patients with painful osteoarthritis of the hip. Compared with racemic ibuprofen half of the daily dose of dexibuprofen shows at least equivalent efficacy. In contrast to pharmacokinetic data, the additional administration of R(-)-ibuprofen in form of racemate does not contribute to the clinical efficacy of racemic ibuprofen. PMID- 10667833 TI - In vivo binding characteristics of phenytoin to serum proteins in monotherapy pediatric patients with epilepsy. AB - AIM: The aim of the present study was to determine the binding characteristics of phenytoin (PHT) to serum proteins in the pediatric population. Binding parameters of PHT to serum proteins in our study were conducted to compare with in vivo or in vitro binding parameters of PHT to serum proteins in adult subjects reported by other investigators. SUBJECTS AND MATERIALS: Serum samples in the study were obtained from 40 pediatric patients (16 male, 24 female) receiving PHT monotherapy. Their age ranged from 1 to 15 years (9.2 +/- 3.6 years, mean +/- SD). The in vivo population binding parameters of PHT to serum proteins and theoretical minimal unbound serum PHT fraction (fu) were determined using an equation derived from the Scatchard equation. RESULTS: The association constant (Ka) was 0.014 l/micromol, while the total concentration of binding sites (n(Pt)) was 747 micromol/l. The number of binding sites per albumin molecule (n) was 1.13, while binding ability (n x Ka) was 0.0161/micromol. The fu was 0.087. The n x Ka is approximately 1.2 times higher in PHT monotherapy adult patients of Pospisil et al. [1992] (i.e. 0.0191 l/micromol) than in all our patients. The association constant is approximately 1.3 times higher in the in vitro study of Monks et al. [1978] (i.e. 0.0186 l/micromol) than in our study, while n is similar between the two studies. The fu in our pediatric patients is similar to the unbound serum PHT fraction in adult patients receiving PHT therapy reported by Richens [1979] (i.e. 0.1). CONCLUSION: Our results suggest that there may be small differences in the binding characteristics of PHT to serum proteins between Japanese pediatric and non-Japanese adult subjects. The unbound serum fraction of PHT in pediatric patients with epilepsy can be assumed to be relatively constant in the therapeutic concentration range of PHT. PMID- 10667834 TI - Analysis of point mutation in exon 2 of CYP2E1 gene in renal cell/urothelial cancer patients in comparison with control population. AB - OBJECTIVE: Genetic polymorphisms of human cytochrome P450s have been implicated to be of importance for susceptibility to different cancers. Recently, a point mutation was found in the exon 2 of the CYP2E1 gene (CYP2E1*2) [Hu et al. 1997]. In order to evaluate a possible link between the point mutation in exon 2 of the CYP2E1 gene and the susceptibility to renal cell/urothelial cancer, we developed a screening method based on the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). MATERIAL: DNA of peripheral white blood cells was isolated from 158 renal cell/urothelial cancer patients as well as from 150 controls. METHOD: Primers for PCR were designed by the Primer 3 release 0.1 program. The PCR yield a product of 215 base pairs (bp), which was digested with the restriction enzyme Hha I. The DNA fragments were separated on a 3% agarose gel stained with ethidium bromide. Restriction enzyme digestion of the PCR product obtained from the wild-type DNA resulted in the appearance of a 66 bp, a 43 bp, a 40 bp, a 39 bp and a 28 bp DNA fragment. In contrast to the wild-type, the digestion of the PCR product from DNA carrying the point mutation resulted in the loss of the 39 bp and 40 bp fragments and the appearance of an additional 79 bp fragment. Therefore, the loss of one Hha I restriction site caused by a single nucleotide exchange is suitable for the identification of the point mutation in exon 2 of CYP2E1 gene. RESULTS: However, we could not detect any point mutation in any of the 158 renal cell/urothelial cancer patients or the 150 controls. The distribution of the point mutation in exon 2 of CYP2E1 gene did not show any difference in renal cell/urothelial cancer patients and controls. CONCLUSION: This might indicate a lack of association between this CYP2E polymorphism (CYP2E1*2) and renal cell/urothelial cancer. PMID- 10667835 TI - Alpha 1-acid glycoprotein (AAG) and serum protein binding of methadone in heroin addicts with abstinence syndrome. AB - OBJECTIVE: To quantify serum protein levels and protein-binding of methadone in vitro in heroin-addicted patients showing objective signs of heroin abstinence. SUBJECTS AND METHODS: Serum samples were obtained from patients (n = 27) hospitalized to participate in a methadone detoxification program and from healthy volunteers (n = 21). The severity of the abstinence syndrome was assessed before blood sampling using a standardized scale. Concentrations of both albumin and alpha1-acid glycoprotein (AAG) were measured in all serum samples. The protein-binding of alpha1-methadone was determined by the ultrafiltration technique and the unbound concentration was measured by liquid scintillation counting. RESULTS: The mean of the AAG concentrations was significantly increased in patients showing signs of withdrawal while the albumin concentrations did not change. Also, the unbound methadone was significantly decreased in this group when compared to the control. A positive correlation (Pearson r = 0.48; p < 0.005) indicates that AAG levels rise during abstinence as the score of withdrawal symptoms increases. Additionally, pooled data from all individuals show the binding of methadone to be related to AAG (r = 0.46; p < 0.05) levels and not to albumin. CONCLUSIONS: The observed changes in protein-binding in abstinence individuals suggest the need for increased dosages of methadone when such patients are treated. Levels of AAG or protein-binding appear to be components of the interindividual variance observed in the response to methadone treatment, hence these variables could be included in future kinetic and dynamic studies. PMID- 10667836 TI - Influence of the administration of amifostine on the pharmacokinetics of 5 fluorouracil in patients with metastatic colorectal carcinoma. AB - A high dose antineoplastic therapy with 5-fluorouracil (5-FU) is associated with severe side effects. It is thought that the cytoprotective drug amifostine can reduce these side effects when it is given prior to a chemotherapeutic course. In this study, the pharmacokinetic parameters of 5-FU are monitored in six patients, who received two chemotherapeutic courses of 2,600 mg/m2 BSA 5FU over 24 h, one course with 700 mg/m2 BSA amifostine prior to the 5-FU infusion and the other without. 20 serum samples were drawn during each infusion time and the 5-FU concentrations were determined by a sensitive and selective GC-MS assay. The statistical analysis of the serum concentrations revealed no significant differences in the pharmacokinetic parameters of 5-FU, whether amifostine is administered or not. The conclusion can be drawn that a reduction of side effects is due to the cytoprotective effect of amifostine and not to a change in the serum concentrations of 5-FU. PMID- 10667837 TI - Specific inhibitors of inducible nitric oxide synthase: efficacy in a rodent model of sepsis. PMID- 10667838 TI - Loss of body cell mass in patients with systemic lupus erythematosus. PMID- 10667839 TI - In vivo lymphoproliferation in the popliteal lymph node (PLN) assay can be inhibited by leflunomide's active metabolite A77 1726. PMID- 10667840 TI - Disease-modifying activity of malononitrilamides, derivatives of leflunomide's active metabolite, on models of rheumatoid arthritis. PMID- 10667841 TI - Cell cycle regulation and inhibition of de novo pyrimidine biosynthesis by leflunomide. PMID- 10667842 TI - Generation of O2- radicals in macrophages can be inhibited in vitro and in vivo by derivatives of leflunomide's primary metabolite. PMID- 10667843 TI - The ability of nicotine to induce glycosaminoglycan release in porcine nasal cartilage explant cultures. PMID- 10667844 TI - Reactive oxygen species generation by mast cells in response to substance P: a NK1-receptor-mediated event. PMID- 10667845 TI - Identification of IL-1 regulated genes in human synovial and gingival fibroblasts by differential display. PMID- 10667846 TI - Cytokines regulate the expression of cellular adhesion molecule in microvascular endothelial cells (MVEC). PMID- 10667847 TI - Granulocyte-colony stimulating factor decreases glycosaminoglycan concentration and increases nitric oxide production in rat articular cartilage. PMID- 10667848 TI - The antiproliferative effect of malononitrilamides (MNAs) on vascular smooth muscle cells is antagonized by exogenous uridine. PMID- 10667849 TI - Ex vivo assays demonstrate potency and selectivity of the COX-2 inhibitor DFP after single dose administration. PMID- 10667850 TI - Trisomy 7 in synovial fibroblasts obtained from arthritic joints. PMID- 10667851 TI - Chronic rejection: the result of uncontrolled remodelling of graft tissue by recipient mesenchymal cells? Data from two rodent models and the effects of immunosuppressive therapies. PMID- 10667852 TI - Induction of COX expression by a tobacco carcinogen: implication in lung cancer chemoprevention. PMID- 10667853 TI - Induction of Tr1 cells: a possible mechanism in the therapeutic action of OM-89. PMID- 10667854 TI - Agonism at melanocortin receptor type 3 on macrophages inhibits neutrophil influx. PMID- 10667855 TI - Selectively 2H-labeled Glu/Asp: application to pKa measurements in Abeta amyloid peptides. AB - Human Abeta peptides have been linked to Alzheimer's disease, and it is hypothesized that formation of amyloid as well as neurotoxicity are important events in the etiology of the disease. Previous studies have shown that the soluble precursor to Alzheimer's amyloid undergoes a pH-dependent folding transition as the self-assembly activity appears, and based upon inter-residue proximities, it was suspected that stabilization of the soluble form might rely upon formation of an intramolecular salt-bridge. However, pKa studies on a model 17-residue Abeta fragment supported an electrostatic model where a solvation imperative for charged side-chain atoms drives the folding process. To explore this model in an active 26-residue fragment as well as the full-length 40-residue Abeta peptide, pKa measurements were performed via 1H and 2H NMR. To overcome issues related to sensitivity and spin system degeneracy, specifically deuterated allyl protected-Fmoc amino acids were synthesized for incorporation into a series of peptides, and a high sensitivity 2H observe NMR probe was constructed. PMID- 10667856 TI - Synthesis and immunological studies of alpha-conotoxin chimera containing an immunodominant epitope from the 268-284 region of HSV gD protein. AB - We have synthesized and characterized new chimeric peptides by inserting an epitope of the glycoprotein D (gD) of herpes simplex virus (HSV) serotype 1 as 'guest' sequence in the 'host' structure of alpha-conotoxin GI, a 13-residue peptide (ECCNPACGRHYSC) isolated from the venom of Conus geographus. The 276-284 region of HSV gD-1 selected for these studies is highly hydrophilic and adopts a beta-turn. The alpha-conotoxin GI also contains a beta-turn in the 8-12 region, stabilized by two disulfide bridges at positions 2-7 and 3-13. Thus, the tetramer sequence of alpha-conotoxin, 8Arg-His-Tyr-Ser12 has been replaced by Asp-Pro-Val Gly (DPVG), identified previously as the epitope core. The syntheses were performed by Fmoc strategy on Rink resin and DTNB or air oxidation were applied for the formation of the first 3-13 disulfide bond in the presence of guanidinium hydrochloride. For the formation of the second disulfide Cys2-Cys7 three different oxidation procedures [iodine in 95% acetic acid, air oxidation in dimethyl sulfoxide/1 M HCl or Tl(tfa)3 in trifluoroacetic acid (TFE)] were compared. The high-performance liquid chromatography purified peptides were characterized by electrospray mass spectrometry and amino acid analysis. The bicyclic HSV-alpha-[Tyr1]-conotoxin chimeric peptide and native alpha-conotoxin GI showed similar circular dichroism spectra in phosphate-buffered saline (PBS) and in a PBS-TFE 1:1 (v/v) mixture, which might suggest that these compounds also share similar secondary structures. In immunologic studies the characteristics of the primary and of the memory immunoglobulin (Ig) M- and IgG-type antibody responses showed that the bicyclic HSV-alpha-[Tyr1]-conotoxin chimera is capable to induce strong antibody responses in C57/Bl/6 mice but was poorly immunogenic in CBA and BALB/c mice. Data obtained with the C57/Bl/6 serum indicate that the polyclonal antibodies recognize the DPVG motif presented in the bicyclic HSV alpha-[Tyr1]-conotoxin and some reactivity was also found with the monocyclic but not with the linear form of the chimera. Results with two IgM type monoclonal antibodies from a bicyclic HSV-alpha-[Tyr1]-conotoxin immunized C57/Bl/6 mouse also point to the specific interaction with the DPVG sequence. Taken together these studies suggest, that the relative intensity of DPVG-specific responses was found to be dependent on the mouse strain and on the conformation of the chimeric molecules. We found that the IgM monoclonal antibodies are able to recognize the linear DPVG sequence, while the majority of IgG antibodies is directed to the same motif in a conformation stabilized by double cyclization. PMID- 10667857 TI - Assignment of the disulfide bonds of Ole e 1, a major allergen of olive tree pollen involved in fertilization. AB - The most prevalent allergen from olive tree pollen, Ole e 1, consists of a single polymorphic polypeptide chain of 145 amino acids which includes six cysteine residues at positions 19, 22, 43, 78, 90 and 131. By using an homogeneous form of the allergen expressed in Pichia pastoris, the array of the disulfide bridges has been elucidated. Specific proteolysis with thermolysin and reverse-phase HPLC separation of the peptides allowed the determination of the disulfide bond between Cys43 and Cys78. Another thermolytic product, which contained three peptides linked by the remaining four cysteines, was digested with Glu-specific staphylococcal V8 protease and the products isolated by reverse-phase HPLC. Amino acid compositions and Edman degradation of the peptide products indicated the presence of the disulfide bonds at Cys19-Cys90 and Cys22-Cys131. These data can help in the analysis of the three-dimensional structure of the protein as well as in studies of its allergenic determinants. PMID- 10667858 TI - Synthesis and characterization of Respiratory Syncytial Virus protein G related peptides containing two disulfide bridges. AB - Respiratory Syncytial Virus (RSV) is the most important cause of bronchiolitis and viral pneumonia in infants and young children. Approximately 100000 children are hospitalized in the USA each year as a result of RSV infections. During the research and development of subunit human Respiratory Syncytial Virus vaccines (hRSV), we have produced numerous synthetic peptides and recombinant proteins containing the four cysteines of the highly conserved central region of the G attachment protein. For several of these disulfide-containing peptides, all possible oxidized isomers were synthesized using various oxidation conditions and resulting in different ratios of isomers. Each isolated isomer was fully characterized by RP-HPLC, FZCE and ES-MS after purification by preparative RP HPLC. The different cysteine pairings were unambiguously established after enzymatic digestion, LC-MS analysis and peptide microsequencing. These synthesis and analytical methods were developed for the characterization on one hand, of recombinant fusion protein BBG2Na which is currently being investigated in advanced clinical phases as a very promising vaccine candidate, and on the other hand, for peptides which were synthesized to be evaluated as conjugate vaccines or as immunochemical tools, after covalent coupling to carrier proteins. Furthermore, these studies allowed us to determine which of the different possible isomers was the most stable and probably the preferred form in native conditions. Finally, the different oxidation and analysis conditions, should be useful for disulfide pairing studies of other peptides and proteins having the same 'xCxxCxxxxxCxxxCx' framework, such as G proteins of non-human RSV strains, developed by other groups as veterinary vaccine candidates for example. PMID- 10667859 TI - Study of the alkylation propensity of cations generated by acidolytic cleavage of protecting groups in Boc chemistry. AB - The alkylation of cysteine residue by different classes of carbonium ions, derived from the cleavage of side chain protective groups in anhydrous HF, was investigated. It was found that side chain protection as beta-2,4-dimethylpent-3 yl ester (Dmp) or 2,4-dimethylpent-3-yloxycarbonyl (Doc) groups resulted in more than seven-fold lower level of alkylated byproducts. This makes Dmp and Doc protection of amino acid side chain during solid phase synthesis particularly valuable in the synthesis of peptides containing cysteine residues or other functional groups prone to alkylation by carbonium ions. PMID- 10667860 TI - Photodynamic effect in lysozyme: a kinetic study in different micellar media. AB - The influence of medium heterogeneity on the kinetics of the photodynamic effect on native protein lysozyme (Lyso), as well as the interaction of protein and the medium, anionic (SDS) micelles, neutral (Triton X-100) micelles and reversed micelles of AOT, were investigated at pH 8. The interaction between Lyso, Triton X-100 and SDS micelles was quantified by determining the respective associations constant (K(Lyso)). Values were 37 M(-1) for Triton X-100 and 514 M(-1) for SDS, indicating that the Lyso molecule binds Triton X-100 micelles effectively and SDS micelles even more strongly. Time-resolved phosphorescence detection (TRPD) indicates that the protein interacts with O2 (1deltag), with overall rate constants of the order of 10(8) M(-1)/S in direct micelles and 10(7) M(-1)/S in reverse micelles. Apparent reactive rate constants for eosin-sensitized photo oxidation (singlet molecular oxygen [O2 (1deltag)]-mediated) of the protein were determined through oxygen uptake experiments for the direct micelles, while the fade in the protein fluorescence spectrum upon sensitized irradiation was used in AOT. The results indicate that the O2 (1deltag) attack on the interior of Lyso on amino acid residues, was more effective in leading to a photo-oxidative reaction in SDS and in Triton X-100 at surfactant concentrations < 1 x 10(-2) M than in a homogeneous solution. However, Lyso reactivity reached a maximum when the concentration of micelles was approximately 1 x 10(-5), the same as the protein concentration In AOT reverse micelles, the quenching rate constants decreased > 75% with respect to water. This effect can be attributed to the decrease in accessibility of the amino acid residues to O2 (1deltag). PMID- 10667861 TI - Relationship between the tertiary structures of mastoparan B and its analogs and their lytic activities studied by NMR spectroscopy. AB - Mastoparan B (MP-B), an antimicrobial cationic tetradecapeptide amide isolated from the venom of the hornet Vespa basalis, is an amphiphilic alpha-helical peptide. MP-B possesses a variety of biological activities, such as mast cells degradation histamine release, erythrocyte lysis and inhibition of the growth of gram-positive and gram-negative bacteria. In order to study the relationship between the structure and the biological activity of MP-B, we used four analogs by replacing amino acids with alanine. Tertiary structures of MP-B and its analogs in 2,2,2-trifluoroethanol (TFE)-containing aqueous solution have been determined by NMR spectroscopy and molecular modeling. The results indicate that [Ala4]MP-B and [Ala12]MP-B with higher hydrophobicity adopt a higher content of amphiphilic helical structures, and have better antimicrobial and hemolytic activities than MP-B. However, [Ala3]MP-B and [Ala9]MP-B with lower hydrophobicity have disordered structures. [Ala3]MP-B and [Ala9]MP-B have low antimicrobial activity and much less hemolytic activity relative to MP-B. It is likely that tryptophan residue in MP-B and appropriate hydrophobicity of MP-B to induce alpha-helical structure is essential for the antibacterial and hemolytic activity of MP-B. This study can aid understanding of the structure-activity relationship of MP-B and to design peptides to possess lytic activity. PMID- 10667862 TI - Conformational properties of a cyclic peptide bradykinin B2 receptor antagonist using experimental and theoretical methods. AB - The solution conformation of the cyclic peptide J324 (cyclo0,6-[Lys0,Glu6,D Phe7]BK), an antagonist targeted at the bradykinin (BK) B2 receptor, has been investigated using experimental and theoretical methods. In order to gain insight into the structural requirements essential for BK antagonism, we carried out molecular dynamics (MD) simulations using simulated annealing as the sampling protocol. Following a free MD simulation we performed simulations using nuclear Overhauser enhancement (NOE) distance constraints determined by NMR experiments. The low-energy structures obtained were compared with each other, grouped into families and analyzed with respect to the presence of secondary structural elements in their backbone. We also introduced new ways of plotting structural data for a more comprehensive analysis of large conformational sets. Finally, the relationship between characteristic backbone conformations and the spatial arrangement of specific pharmacophore centers was investigated. PMID- 10667863 TI - Analysis of binding sites and efficacy of a species-specific peptide at rat and human neurotensin receptors. AB - We have developed a neurotensin analog, L-[3,1'-naphthylalanine11]NT(8-13), NT34, that can distinguish between rat and human neurotensin receptors, and exhibits more than a 100-fold difference in binding affinities and a 60-fold difference in functional coupling to phosphatidylinositol turnover. Using cells transfected with different numbers of the appropriate receptors, we measured the changes in phosphatidylinositol production, and then evaluated the efficiency of receptor effector coupling based on Furchgott's design. The binding of NT34 at both rat and human neurotensin receptors stably expressed in CHO-K1 cells was to two sites, while the binding of NT was to one site. At the rat receptor the equilibrium dissociation constant (Kd) for NT34 at the high-affinity site was 0.058 nM, while that at the low-affinity site was 3.1 nM. For the human receptor at the high-affinity site, the Kd for NT34 was 18 nM, while that at the low affinity site was 180 nM. For both species the percentage of receptors representing the high-affinity site was approximately 60-70% with 30-40% at the low-affinity site. We derived agonist dissociation constants (Ka) for NT and NT34, which suggest that for NT34, the low-affinity site is functionally coupled to phosphatidylinositol turnover. Finally, we compared the relative efficacies of both compounds and found that NT34 was about 2-fold and 4-fold more efficacious than NT in stimulating phosphatidylinositol turnover in rat and human NT receptors, respectively. PMID- 10667864 TI - Chemical synthesis and receptor binding of catfish somatostatin: a disulfide bridged beta-D-Galp-(1-->3)-alpha-D-GalpNAc O-glycopeptide. AB - The glycopeptide hormone catfish somatostatin (somatostatin-22) has the amino acid sequence H-Asp-Asn-Thr-Val-Thr-Ser-Lys-Pro-Leu-Asn-Cys-Met-Asn-Tyr-Phe-Trp Lys-Se r-Arg-Thr-Ala-Cys-OH; it includes a cyclic disulfide connecting the two Cys residues, and the major naturally occurring glycoform contains D-GalNAc and D Gal O-glycosidically linked to Thr5. The linear sequence was assembled smoothly starting with an Fmoc-Cys(Trt)-PAC-PEG-PS support, using stepwise Fmoc solid phase chemistry. In addition to the nonglycosylated peptide, two glycosylated forms of somatostatin-22 were accessed by incorporating as building blocks, respectively, Nalpha-Fmoc-Thr(Ac3-alpha-D-GalNAc)-OH and Nalpha-Fmoc-Thr(Ac4-beta D-Gal-(1-->3)-Ac2-alpha-D-GalNAc)-O H. Acidolytic deprotection/cleavage of these peptidyl-resins with trifluoroacetic acid/scavenger cocktails gave the corresponding acetyl-protected glycopeptides with free sulfhydryl functions. Deacetylation, by methanolysis in the presence of catalytic sodium methoxide, was followed by mild oxidation at pH 7, mediated by Nalpha-dithiasuccinoyl (Dts) glycine, to provide the desired monomeric cyclic disulfides. The purified peptides were tested for binding affinities to a panel of cloned human somatostatin receptor subtypes; in several cases, presence of the disaccharide moiety resulted in 2-fold tighter binding. PMID- 10667865 TI - Changes inside Academic Medicine: why not a real cure instead of a band-aid? PMID- 10667866 TI - Kampo medicine training in Japanese medical schools. PMID- 10667867 TI - Book clubs in residents' education. PMID- 10667868 TI - PubMed Central and the new publishing landscape: shifts and tradeoffs. PMID- 10667869 TI - Protecting privacy without shackling providers. PMID- 10667870 TI - Healthy people 2010: setting the nation's public health agenda. PMID- 10667871 TI - Three strategies used by academic health centers to expand primary care capacity. AB - The growth of managed care in the late 1980s and early 1990s severely disadvantaged academic health centers (AHCs). The reliance on primary care gatekeeping and selective contracting by managed care plans were two contributing factors. Because most AHCs had only a modest primary care capacity, they were understandably concerned about their strategic positions. Thus, many felt it was essential to expand their primary care capacities to ensure downstream referrals, to improve contract negotiations with third parties, and to permit assumption of risk for defined populations. Among the different approaches used, three principal strategies emerged for the expansion of the primary care capacity of AHCs: (1) the "assembly strategy," in which many AHCs recruited new generalist faculty into existing clinical departments; (2) the "acquisition strategy," in which AHCs purchased established primary care practices in the community; and (3) the "affiliation strategy," in which some AHCs affiliated with primary care physicians in the community and formed networks of academic and community physicians. For each of these approaches, the author reviews the relative merits and disadvantages, and analyzes why some AHCs' original assumptions about the imperative for increasing primary care capacity may have been spurious. He concludes that recent marketplace and regulatory changes may make it less necessary for AHCs to secure substantial primary care bases in the future. PMID- 10667872 TI - Literature and medicine: origins and destinies. AB - Literature and medicine is a flourishing subdiscipline of literary studies that examines the many relations between literary acts and texts and medical acts and texts. The author examines the historical connections between these two fields and suggests that the growth and decline in medicine's attentiveness to the power of words can be used as a marker for medicine's degree of attentiveness to the individual patient's predicament. The recent explosive growth in medicine's interest in literature and narrative is taken as evidence that medicine's swing toward the reductionist and away from the narrative has ended. Patients and doctors have reason to await the return swing of the pendulum-if not the turn of the spiral-toward a medicine that is both technologically and narratively competent. PMID- 10667873 TI - Assessing the operating efficiencies of teaching hospitals by an enhancement of the AHA/AAMC method. American Hospital Association/Association of American Medical Colleges. AB - In the ongoing effort to control costs, comparisons among hospitals' efficiency levels, if valid, can help identify "best practices" across institutions and uncover situations that need corrective intervention. The authors present an extension of the "adjusted cost per equivalent discharge" approach, which incorporates case-mix-severity differences, regional labor cost differentials, and inpatient/outpatient mix, but does not take into account such factors as the differences in hospital sizes, extents of the teaching mission, or quality of care delivered. The alternative approach yields information that suggests where an institution's total operating costs might be reduced with no change in any of the hospital's outputs or operating environment, through comparison with a "peer group" of other hospitals, matched according to the subject hospital's number of beds, the quality of care the hospital delivers, the extent of medical education carried out, the level of case-mix-adjusted discharges, and outpatient activities. A difficulty with this approach (as with others) is that measurement of some of the additional facets (e.g., quality of care) is still evolving, so its main contribution at this time is to provide a construct and method capable of incorporating these important added considerations. Hospital rankings achieved by applying the current and alternative approaches to a real set of teaching hospitals operating in FY 1987 are compared. While the rankings produced by the two approaches are loosely similar, the authors show that some significant differences do appear and can be at least partially explained by the incorporation of the additional factors mentioned above. PMID- 10667874 TI - An integrated residency in internal and preventive medicine. AB - The importance of preventive and population-based principles in clinical practice is widely acknowledged. The challenge of imparting these principles in either undergraduate or postgraduate medical education has, however, not been fully met. The necessary skills are provided comprehensively by preventive medicine residency programs, but at the expense of clinical training. Sequential residencies in primary care and preventive medicine, the currently available means of obtaining thorough preparation in both clinical and population-based principles, represent an inefficient, generally unappealing, and non-integrated approach. In response to these concerns, and in an effort to make preventive medicine training appeal to a wider audience, the authors developed and implemented a residency program fully integrating internal and preventive medicine. The program meets, and generally exceeds, the requirements of both specialty boards over a four-year period. The program provides extensive training in clinical, preventive, and public health skills, along with case management and cost-effective care, conferring the MPH degree and leading to dual board eligibility. The model is ideally wed to the demands of the modern health care environment in the United States, is extremely attractive to applicants, and may warrant replication both to train academic and administrative leaders and to raise the standards of preventive and public health practice in primary care. PMID- 10667875 TI - A thorough pulmonary exam and other myths. PMID- 10667876 TI - A randomized controlled study of brief interventions to teach residents about domestic violence. AB - PURPOSE: To test an educational intervention regarding domestic violence. METHOD: Residents beginning their training in 1995 or 1996 were randomly assigned to attend, at their hospital orientation, either a 20-minute session emphasizing the importance of screening for domestic violence or a session on an unrelated topic. RESULTS: Seventy-one percent of the residents in the experimental group diagnosed domestic violence; 52% in the control did so (RR, 1.35; 95% CI, 0.96-1.90; p = .07) in the nine to 12 months following the intervention. Rates of diagnosis differed by specialty (p < .01): 100% family practice, 90% emergency medicine, 80% obstetrics-gynecology, 63% pediatrics, 47% internal medicine, 0% surgery. Change in knowledge was assessed in 1996; significant improvement was noted (p = .002). CONCLUSION: An intervention about domestic violence conducted at orientation for residents improved the rate of diagnosis of domestic violence. While the improvement was not statistically significant in this case, the intervention was brief and harmless. Other institutions should consider this kind of brief intervention. PMID- 10667877 TI - Teaching medical students about continuity of patient care. AB - This article reports on medical schools' longitudinal primary care ambulatory programs and five themes identified by educators as to why it is important for students to experience continuity of patient care. It briefly describes methods used to expose students to this concept, which is so basic to primary care practice. PMID- 10667878 TI - Evaluation of Web-based computer-aided instruction in a basic science course. AB - PURPOSE: To demonstrate the applicability of server statistics, in combination with user surveys, to evaluate utilization of Web-based computer-aided instruction (CAI) in the undergraduate medical curriculum. METHOD: Individual user surveys with students' names provided information about computer literacy prior to the course and use of CAI during the course. Utilization of specific web based CAI developed for the course was recorded by server software and the daily logs correlated with course content. Regression analyses were used to measure correlation of server access logs of individual students versus information from user surveys and performances in the course based on in-course examinations. RESULTS: There was no correlation between computer literacy of students at matriculation and their subsequent levels of use of CAI in the curriculum. Utilization of CAI developed for specific course objectives coincided closely with course content, which is an indication of the effectiveness of the applications in achieving their curricular objectives. In contrast, student use of tutorials coincided most closely with in-course examinations. Students' responses to surveys were generally substantiated by server statistics, but discrepancies were sufficiently large (10% to 20%) to call into question the validity of these surveys. Significant differences in CAI utilization correlated with the performances of students in the course. CONCLUSIONS: This study demonstrates an important advantage of web-based applications to collect and evaluate CAI utilization efficiently and objectively at both the level of the class and the level of the individual student. PMID- 10667879 TI - Student performances on Step 1 and Step 2 of the United States Medical Licensing Examination following implementation of a problem-based learning curriculum. AB - PURPOSE: To examine students' performances on Step 1 and Step 2 of the United States Medical Licensing Examination (USMLE) following the implementation of a problem-based learning curriculum. METHOD: Performances on Step 1 of the USMLE for four classes at the University of Missouri-Columbia School of Medicine that completed a new problem-based learning curriculum (1997, 1998, 1999, and 2000) were compared with those of the last two classes to learn in the traditional curriculum (1995 and 1996). Performances on Step 2 of the USMLE for the classes of 1997, 1998, and 1999 were also compared with those of the classes of 1995 and 1996. The authors analyzed matriculation data (GPAs and MCAT scores) for all six classes. They compared all data with those of U.S. and Canadian first-time USMLE takers. RESULTS: The mean scores were higher on USMLE Step 1 for classes in the problem-based learning curriculum than for classes in the traditional curriculum. The mean scores for Step 2 were above the national mean for classes in the revised curriculum and below the national mean for classes in the traditional curriculum. The admission profiles of these classes were essentially the same before and after the change in curriculum. CONCLUSIONS: Major PBL revisions of the curriculum did not compromise the performances of medical students on the licensing examinations; in fact, they may have contributed to higher scores. PMID- 10667880 TI - A pilot survey study to define quality in residency education. AB - PURPOSE: To begin to define indicators of quality in internal medicine residency training. METHOD: In 1995, through a modified Delphi process, the Association of Program Directors in Internal Medicine's Research Committee developed a questionnaire containing 44 items (34 process and ten outcome indicators). The survey was mailed to all 418 internal medicine program directors and a convenience sample of medical residents. RESULTS: Responding at a rate of 78% (326), program directors rated several indicators as important. These included such faculty characteristics as stability, completeness, supervision, clinical skills, and teaching commitment; institutional support; amount of resident evaluation and feedback; encouragement of lifelong learning; and ability to meet its program goals. There was strong agreement between faculty and residents (r = 0.91). Items rated less important included graduates' selecting academic or generalist careers, residents' caring for elective cardiac catheterization patients, resident community service, training minorities and women, and faculty research. CONCLUSION: These results demonstrate the diversity of opinion of what defines quality in residency education and the emphasis placed on process rather than outcome indicators. To be valid, future endeavors must include all those with a stake in graduate medical education, including accrediting bodies, future employers, and patients. PMID- 10667881 TI - Program directors' perspectives on federally funded fellowship training in primary care research. AB - PURPOSE: To describe the organization, models of training, and institutional impact of National Research Service Award fellowship programs in primary care research. METHOD: Survey of 25 directors of currently-funded and former training sites. RESULTS: Twenty-four program directors (96%) completed the survey. Programs allocated 39% of fellows' time to course work leading to an advanced degree or other didactic instruction, and 40% of time to the conduct of research. Collaborations with other training programs within the institution occurred at 83% of sites. Programs commonly (54%) or exclusively (42%) relied on a research model of "early research independence" in which the fellow defined an area of research interest, rather than an "apprenticeship" model in which the fellow worked in a senior investigator's research area. These programs enriched the local academic environment, but required extensive financial subsidies. The high costs of training often had adverse impacts on recruitment and other components of the training process. CONCLUSION: Research training programs in primary care often substitute acquisition of advanced degrees for early immersion in research. The "early independence" model of research differs from fellowships in the medical specialties, and requires further study to assess its effectiveness. The need to subsidize training costs poses substantial problems for the institutions that host these fellowship programs. PMID- 10667882 TI - Using early clinical experiences to integrate quality-improvement learning into medical education. AB - Health care providers are delivering care in an increasingly complex environment; this requires that providers develop new competencies to better understand their work and to design changes that can help them succeed. Recognizing these new educational requirements, Dartmouth Medical School created a model two-pronged program for teaching quality improvement to its medical students. The goal of the program is to provide students with an active learning experience as well as an education in the theory and application of continuous quality improvement. The program includes two educational experiences: one curriculum is for all medical students and the other is for selected, highly motivated students. The first curriculum is incorporated in Dartmouth's required "On Doctoring" course, in which students spend time with community-based physician preceptors. The quality improvement curriculum is designed around an improvement project developed at the students' preceptor sites. The second curriculum for students with a special interest in quality improvement is offered as an elective summer program between the first and second years of medical school. Working in groups of two, students identify an area for improvement within a preceptor's practice, assist the practice in articulating an improvement plan, help implement that plan, and write up their experiences. The authors describe the two curricula, factors associated with their successful implementation, and lessons learned. PMID- 10667883 TI - Eva's stories: recognizing the poverty of the medical case history. AB - The medical case history is a proven tool for approaching the question "What is wrong with this person?" Its virtues, however, can become vices, in part as a consequence of the dehumanizing flight from sensitive subjectivity to sanitized objectivity, from human interest to "science." The case history, because it is so useful and effective, is not likely to be profoundly altered in the future, but medical educators can make themselves and their students more aware of the serious flaw in this form of discourse, i.e., the erasure of the unique individual from his or her disease. The exercise of asking medical students to abstract case histories from richly written short stories, novels, plays, or operas might heighten students' recognition of the poverty of the medical case history. To illustrate this idea, the story of Eva, a dying woman, is presented initially as a typical medical case history; it is then contrasted with excerpts from a novelist's narrative of Eva's life. PMID- 10667884 TI - Teaching medical students clinical reasoning skills. AB - To be an effective clinician, a physician must excel at clinical problem solving. Nevertheless, few physicians have been specifically taught clinical reasoning skills during their medical training. This paper describes the format used to teach clinical problem solving to second-year medical students at the University of Connecticut School of Medicine. PMID- 10667885 TI - Glaucoma screening as a vehicle for providing continuing medical education. AB - During a free screening at a major medical meeting, volunteer ophthalmologists educated attendees about glaucoma. A questionnaire was given to participants before and six months after the screenings to evaluate changes in their knowledge of glaucoma risk factors; the results showed a significant and enduring improvement (p < .02) in respondents' knowledge about glaucoma. PMID- 10667886 TI - Integrating immunotoxicity with effects on other biological systems in preclinical safety evaluation: a perspective. PMID- 10667887 TI - Endocrine and neurological adverse effects of the therapeutic interferons. AB - There is experimental evidence that the nervous central and the neuroendocrine systems can influence the immune system, which can in turn influence the brain activity. Endogenous cytokines are known to play a critical role in the pathophysiology of many diseases. The recently acquired experience on the adverse effects of therapeutic cytokines, particularly neurological and endocrine adverse effects, are further illustrative of these interferences. Interferons-alpha have been used in thousands of patients, so that the information accumulated with this group of closely related products is essential to delineate the potential and severity for non-immunological, but largely immune-mediated adverse effects to develop in patients treated with immuno-activating agents. PMID- 10667888 TI - Hepatic drug metabolism and immunostimulation. AB - When host defence mechanisms are stimulated there is a concomitant decrease in cytochrome P450 based drug biotransformation and elimination. This has resulted in a number of clinically important unwanted drug responses in patients with infections or inflammatory responses. The loss in cytochrome P450 is predominantly an effect at the level of the gene expression and the majority of enzyme forms examined to date are involved. Although the effect occurs predominantly in the liver it has been recently shown that inflammatory responses in the brain also cause a loss of the same enzyme forms in that organ. The loss of cytochrome P450 in the brain in response to localised inflammation is accompanied by a similar loss in the liver. The decrease of cytochrome P450 and its dependent drug biotransformation is of concern whenever drugs are used in patients with infections or disease states with an inflammatory component. PMID- 10667889 TI - Past, present and future of psychoneuroimmunology. AB - Psychoneuroimmunology was for the first time comprehensively described about 20 years ago. The influence of mental status on the course and outcome of a number of diseases, however, was suspected a long time before. Also the links between mental affective disorders and the immune status were repeatedly suggested. The authors in this paper shortly reviewed the most important clinical as well as experimental evidence which at present strongly supports the concept of a close and bidirectional communication between central nervous, neuroendocrine and immune systems. The most important anatomical, physiological as well as pharmacological experimental data, which were obtained by the authors during 20 years of research in this field, are presented. The data strongly suggest that in the very next future we will not only better understand a very complex communication between mind and body, but also completely new types of compounds might become available. PMID- 10667890 TI - Central/peripheral nervous system and immune responses. AB - Maintenance of health is dependent on numerous regulatory interactions between organ systems. This review discusses interorgan communication between the nervous, endocrine, and immune systems and environmental and genetic influences on this neuroendocrine immune circuitry. Stresses of multiple types, including psychological and exposure to chemicals and infectious agents, may combine to enhance neuroimmunotoxicology. Altered nervous system functions can alter immunity which could result in exacerbation of infections, cancers or other immune-associated problems. Inversely, aberrant immune system activities could lead to pathologies associated with altered nervous activities, such as Alzheimer's disease, chronic fatigue, or multiple sclerosis. The nervous, endocrine and immune circuitry is multi-directional, and a chemical, physical or emotional stress could upset the homeostasis. PMID- 10667891 TI - Epidermal cytokines in experimental contact dermatitis. AB - Topical exposure to a variety of xenobiotics may result in irritant as well as allergic contact dermatitis both in rodents and in humans. Despite their induction by different mechanisms, they cannot be differentiated by macroscopic appearance and, by histological examination they are both generally characterized by a perivascular mononuclear cell infiltrate and capillary hyperpermeability. Recently, cytokines, a family of inducible glycoproteins that play a pivotal role in immune and inflammatory reactions, have been identified as useful tools for differentiation of irritant and allergic contact dermatitis. In this article the role of cytokines in the development and differentiation of irritant and allergic contact dermatitis is discussed. PMID- 10667892 TI - Approaches to the identification and recording of findings in the lymphoreticular organs indicative for immunotoxicity in regulatory type toxicity studies. AB - Recent validation studies showed that histopathological examination of the hematopoietic and lymphoreticular system is one of the most sensitive tools in the evaluation of non-specific immune stimulatory or immune suppressive effects and hazard identification of potential immunotoxicity in routine safety tests. Most immunotoxic effects have a classical dose-response relationship and an immediate effect (i.e. do not need an induction phase). The findings associated with a specific immunomodulatory response are generally not detected morphologically in routine sections of the immune system in safety studies, but may be detected, because of their effects on other organs such as skin (contact dermatitis) or joints and kidneys (immune complex deposits). Careful detailed examination of the immune system may give valuable clues for the possible mechanism of action of the test material, as was also demonstrated in these validation studies. PMID- 10667893 TI - Genitourinary medicine and sexually transmitted infections in the 21st century. PMID- 10667894 TI - A retrospective study of neutropenia in HIV disease. AB - In aiming to define the characteristics of HIV-infected subjects developing neutropenia and describe the causes, features and effects of neutropenia we undertook a retrospective study in a dedicated HIV unit in London, UK. Two hundred and forty-four patients with documented neutropenia, defined as absolute neutrophil count (ANC) < 1,000/mm3, during a 12-month period were studied. First neutropenia occurred at a median CD4 count of 30 cells/mm3. Low CD4 count was associated with longer episodes of neutropenia with a more profound nadir. Two thirds of episodes lasted less than 2 weeks. ANC nadir was < 500 cells/mm3 in 45% of episodes. Infections were most frequent in patients with profound but brief neutropenic episodes. Neutropenia was generally mild, short-lived and associated with late-stage disease. However, profound neutropenia did develop suddenly in some patients with no prior history of neutropenia (in 13% first neutropenic ANC recorded was < or =500 cells/mm3), and at CD4 count > 200 cells/mm3. Most patients were receiving multiple myelosuppressive therapies. Infection was associated with brief, profound neutropenia. PMID- 10667895 TI - Psychosocial impact of type-specific herpes simplex serological testing on asymptomatic sexual health clinic attendees. AB - The usefulness of type-specific testing for herpes simplex virus type 2 (HSV-2) is much debated with proponents arguing for likely change in the sexual behaviour of asymptomatic carriers and opponents suggesting that testing may have significant adverse psychological impact while not promoting behaviour change. In the present study we examine the impact of HSV-2 serological testing on psychological well-being, self-esteem, anxiety, sexual self-perceptions and sexual practices among a sample of 180 clients of a sexual health clinic. Of the participants, 21 (11.7%) were HSV-2 positive at entry to the study. No adverse psychological consequences of a positive test were observed among those participants followed for 3 months (n=124) or for 6 months (n=97). There was little significant change observed in sexual behaviour although a general but not significant pattern of decreased sexual behaviour with casual partners was observed among participants who tested HSV-2 negative. While supportive of the value of HSV-2 testing, these findings require replication in larger samples and different populations. PMID- 10667897 TI - The effect of modest monetary incentives on follow-up rates in sexually transmitted disease studies. AB - The purpose of the study was to determine if follow-up rates in sexually transmitted disease (STD) research could be improved by offering modest monetary incentives. Women aged 14-34, infected with Chlamydia trachomatis, and enrolled in either of 2 studies between May 1995 and January 1999 were included. Beginning in March 1996 participants were offered a $20 incentive to return to both the one and four-month follow-up visits. Data were analysed using polychotomous logistic regression. Of 962 women followed, the majority (74.7%) received monetary incentives and 66% returned for at least one visit. Women who received the incentive were more likely (OR 1.9, CI 1.2-2.9) to return for either one or both of their follow-up visits after adjusting for interviewer and the months of work experience of that interviewer. Age and method of birth control were not associated with return rates. The study provides evidence that modest monetary incentives can improve follow-up rates. PMID- 10667896 TI - Going underground and going after women: trends in sexual risk behaviour among gold miners in South Africa. AB - This paper examines trends in risk behaviour among South African gold miners, a population with an estimated HIV prevalence of 10-20%. The study is based on a 1995 and 1997 survey of a random sample gold miners in the town of Welkom, South Africa. The results show that the percentage of miners who perceived they were likely to contract HIV increased from 33% in 1995 to 35% in 1997 (P<0.01). The percentage who had 4 or more partners in the past year decreased from 25% to 13% (P < 0.01), and the percentage whose last sexual partner was their spouse increased from 56% to 70% (P<0.01). Condom use in last intercourse with a spouse increased from 18% to 26% (P < 0.05). Condom use with other partners was considerably higher (67%), but did not increase significantly from 1995. The most likely contributors to this behaviour change were the AIDS awareness programmes implemented by the mining industry and the behaviour change communications of a condom social marketing campaign targeted at miners and commercial sex workers in the mining community. PMID- 10667898 TI - Seronegative HIV-2 carriers in India. AB - The discordant cases of seronegative, but culture and proviral HIV-2 DNA positive were found in Mumbai, India. This was corroborated by the successful isolation of HIV-2-RNA in culture medium, HIV-2 cDNA sequence determination and the detection of the antigen. The sequence of the isolated HIV-2 genomic RNA does not seem to be altered to the extent that the change will alter antibody binding. Furthermore, antibody from the same individual (even at 8 months from initial sampling) from whom HIV-2 was isolated did not react with the antigen of this strain. Those evidences imply that extremely low or non-production of the antibody may be due to suboptimal immune stimulation due to extremely slow HIV-2 replication. This low virus-load may be responsible for the negative antibody results in the HIV-2 carriers. PMID- 10667899 TI - Secondary HIV transmission rates in a mixed-gender sample. AB - Information about the sexual behaviour of HIV-infected individuals is needed to predict the course of the sexually transmitted HIV epidemic in the US. The present study provides model-based estimates of the secondary transmission rate (i.e. the number of infections expected among the sex partners of already infected individuals) for a sample of HIV-positive persons in Atlanta. A mathematical model was used to estimate the secondary transmission rate of HIV infection for a sample of HIV-positive men and women in Atlanta, based on their self-reported sexual behaviour, extrapolated over a 15-year horizon. Separate rates were calculated for different transmission routes, including: from women to men-who-have-sex-with women (MSW) and from men-who-have-sex-with-men (MSM) to other MSM. Sensitivity analyses were conducted to assess the impact of different parametric and modelling assumptions. Restricted to the sub-sample that reported transmission risk behaviours, the mean number of secondary infections was 0.14 for transmission from women to MSW; 0.31 for transmission from MSW to women; and 0.84 for MSM to MSM transmission. Bisexual men were at especially high transmission risk, with 1.59 and 0.54 secondary infections expected among their male and female partners, respectively. The main analysis indicates that, in this sample, each current infection will lead to fewer than one future infection for all groups other than bisexual men, which suggests that the epidemic is contracting in this community, although this analysis cannot rule out the possibility of a growing epidemic among MSM. This method can be used to identify groups at high risk for HIV transmission and thereby to better target HIV prevention resources. PMID- 10667900 TI - Disease prevalence in women attending the STD clinic in Mumbai (formerly Bombay), India. AB - Our objectives were to determine the prevalence of Neisseria gonorrhoeae and its association with other STD causing organisms. Three hundred and thirty-six consecutive women (female sex workers (FSWs) and married contacts), attending a sexually transmitted disease (STD) clinic in Mumbai, were screened for N. gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis. Per speculum examination was performed and clinical signs were recorded. Symptoms perceived by the women were also recorded. The mean age for married contacts, FSWs and gonorrhoea-positive women was 27.9, 29.7 and 27.5, respectively. 9.7% of the women were positive for N. gonorrhoeae, 23.2% were chlamydia-positive and 5.9% had trichomoniasis. N. gonorrhoeae was isolated more frequently from FSWs as compared to the married contacts. The prevalence of HIV was significantly higher among women with multiple sex partners (FSWs) (P<0.001). Gonococcal infection is significantly associated with the presence of HIV. A significant association between sexual habits and prevalence of gonorrhoea, trichomoniasis and HIV was observed. The prevalence of gonorrhoea over 1988 to 1996 remained approximately the same. PMID- 10667901 TI - Increase in plasma IL-10 levels and rapid loss of CD4+ T cells among HIV-infected individuals in south India. AB - Increased levels of immune activation among HIV patients from developing countries are believed to accelerate and/or enhance the shift to a Th2 cytokine environment, which in turn may result in a more rapid progression to AIDS. In support of this hypothesis, we present data from a cohort of 35 HIV+ individuals in southern India. Among asymptomatic individuals in this cohort, a dramatic increase in plasma interleukin (IL)-10 coincided with rapid decrease in CD4 counts and progression to AIDS. Serum IL-10 levels were significantly higher after 6 months of follow up (P=0.01), while CD4 counts declined at a rate of 280 cells/ul per year, roughly 3 times the rate of decline reported for HIV+ asymptomatic subjects in developed countries. Changes in serum IL-10 levels and CD4 counts fell short of statistically significant correlation (P=0.1). Among AIDS patients in this cohort, the mean period from diagnosis of AIDS to death was <5 months and is in agreement with an earlier report of rapid progression in India. PMID- 10667902 TI - The impact of zidovudine on dementia-free survival in a population of HIV positive men and women on antiretroviral therapy. AB - Our objective was to characterize the effect of zidovudine therapy on AIDS dementia complex (dementia) free survival among HIV-infected men and women in a population-based cohort with free access to antiretroviral therapy in the province of British Columbia. Time to diagnosis of dementia among individuals was examined on the basis of zidovudine duration, CD4+ cell count at first treatment, gender, and transmission group [men having sex with men (MSM), intravenous drug users (IDU), heterosexuals]. We restricted the analysis to subjects with CD4+ cells counts within 12 months prior to treatment start date. Among 641 participants eligible for analysis, median duration of follow-up was 3.6 years, under which 86 (9.3%) events of dementia occurred. Participants were less likely to develop dementia with: increased zidovudine exposure (OR=0.26, 95% CI: 0.14 0.49), at least 260 CD4+ cells/mm3 (median) (OR=0.52, 95% CI: 0.34-0.78), and MSM risk group (OR=0.57, 95% CI: 0.35-0.94). Those infected through heterosexual contact had an increased risk (RR=2.04, 95% CI: 1.02-4.07). Using Cox's proportional hazards model, controlling for CD4+ cell count at treatment start date, independent predictors of dementia-free survival were: duration of zidovudine (OR=0.28, 95% CI: 0.15-0.52) and MSM transmission group (OR=0.61, 95% CI: 0.37-1.00). In this observational treatment cohort, factors associated with dementia-free survival include duration of zidovudine (AZT) therapy and MSM transmission group. It is not clear from these data whether the AZT protective effect is exclusive to this agent or whether other therapies might offer a similar protective effect. PMID- 10667903 TI - Perceived compliance with AZT dosing among a sample of African-American drug users. AB - The purpose of this report was to present findings from a pilot study conducted to explore the associations between sociodemographic, drug use, and health belief factors and perceived compliance with zidovudine (AZT) among African-American drug users. Data were collected in Washington, DC, USA from individuals who were African-American; were recent or current drug injectors or crack smokers; were HIV-seropositive, and were receiving treatment for HIV infection. Participants were recruited through local organizations that provide services to HIV-infected persons. Participants were interviewed using a questionnaire that solicited sociodemographic, lifetime and current drug use, current sexual behaviours, health status, HIV and drug treatment history, and health belief data. Analyses were limited to individuals currently using an illicit substance and who had received AZT during their medical treatment. Parametric (Pearson's r) and nonparametric (Spearman's rho) statistics were used to assess correlations between perceived compliance with AZT dosing and independent variables. As the study was intended to be both descriptive and exploratory, the level of statistical significance was set at 0.10, rather than the customary 0.05. Antiretroviral medications recognized and recalled by participants are presented. The most commonly recalled medication was AZT. Slightly less than one-third of participants reported being completely compliant with an AZT regimen. Perceived compliance was found to be negatively associated with 5 variables: age, homelessness, number of injections in the previous 30 days, trading sex for drugs, and the perception that AIDS is no longer a serious disease since the development of new antiretroviral medications. Intensity of feelings of joy, fear, and the belief that taking more anti-HIV medications would result in better health were found to be positively correlated. Bivariate associations between perceived compliance and sociodemographic, drug use, sexual behaviour, and health belief variables suggest further avenues of study and potential points for intervention to increase compliance with antiretroviral medications among racial/ethnic minority drug users receiving treatment for HIV infection. PMID- 10667904 TI - Pneumocystis carinii pneumonia following discontinuation of primary prophylaxis despite highly active antiretroviral therapy. PMID- 10667905 TI - Sexually transmitted infections and cervical neoplasia. PMID- 10667906 TI - Update on the experimental human model for chancroid infection. PMID- 10667907 TI - Ciprofloxacin-resistant gonorrhoea. PMID- 10667908 TI - Antibiotic prophylaxis and suction termination of pregnancy. PMID- 10667909 TI - Termination of pregnancy, chlamydia and contact tracing. PMID- 10667910 TI - A proposed BAT-26 germline polymorphism. PMID- 10667911 TI - The role of NAC in amyloidogenesis in Alzheimer's disease. PMID- 10667912 TI - Additional literature on "vasculogenic mimicry" not cited. PMID- 10667913 TI - Surface antigen CD98(4F2): not a single membrane protein, but a family of proteins with multiple functions. PMID- 10667914 TI - Effect of calcium and calcium channel blockers on transient outward current of F76 and D1 neuronal soma membranes in the subesophageal ganglia of Helix aspersa. AB - Twin-electrode voltage-clamp techniques were used to study the effect of calcium and calcium channel blockers on the transient outward current in isolated F76 and D1 neurones of Helix aspersa subesophageal ganglia in vitro (soma only preparation with no cell processes). On lowering extracellular Ca(2+) concentration from 10 to 2 mm or removing extracellular calcium from the bathing medium, the threshold for this current shifted in a negative direction by 11. 5 and 20 mV, respectively. On the other hand, increasing the extracellular Ca(2+) concentration from 10 to 20 and to 40 mm shifted the steady-state inactivation curves in positive directions on the voltage axis by 7 and 15 mV, respectively. Upon application of calcium channel blockers, Co(2+), La(3+), Ni(2+) and Cd(2+), transient potassium current amplitude was reduced in a voltage-dependent manner, being more effective at voltages close to the threshold. The current was elicited even at a holding potential of -34 mV. The specific calcium channel blockers, amiloride and nifedipine did not shift the activation and steady-state inactivation curves and did not reduce the transient outward current amplitude. It was concluded that the transient outward current is not dependent on intracellular Ca(2+) but that it is modulated by Ca(2+) and di- and trivalent ions extracellularly. The effects of these ions are very unlikely to be due to a surface charge effect because the addition of La(3+) (200 microm) completely reverses the shift in a hyperpolarizing direction when the extracellular Ca(2+) concentration was reduced from 10 to 1 mm and additionally shifts the kinetics further still in a depolarizing direction. The responses seen here are consistent with a specific effect of di- and trivalent ions on the transient outward current channels leading to a modification of gating. PMID- 10667915 TI - Influence of a transmembrane protein on the permeability of small molecules across lipid membranes. AB - The influence of the nonchannel conformation of the transmembrane protein gramicidin A on the permeability coefficients of neutral and ionized alpha-X-p methyl-hippuric acid analogues (XMHA) (X = H, OCH(3), CN, OH, COOH, and CONH(2)) across egg-lecithin membranes has been investigated in vesicle efflux experiments. Although 10 mol% gramicidin A increases lipid chain ordering, it enhances the transport of neutral XMHA analogues up to 8-fold, with more hydrophilic permeants exhibiting the greatest increase. Substituent contributions to the free energies of transfer of both neutral and anionic XMHA analogues from water into the bilayer barrier domain were calculated. Linear free-energy relationships were established between these values and those for solute partitioning from water into decadiene, chlorobutane, butyl ether, and octanol to assess barrier hydrophobicity. The barrier domain is similar for both neutral and ionized permeants and substantially more hydrophobic than octanol, thus establishing its location as being beyond the hydrated headgroup region and eliminating transient water pores as the transport pathway for these permeants, as the hydrated interface or water pores would be expected to be more hydrophilic than octanol. The addition of 10 mol% gramicidin A alters the barrier domain from a decadiene-like solvent to one possessing a greater hydrogen-bond accepting capacity. The permeability coefficients for ionized XMHAs increase with Na(+) or K(+) concentration, exhibiting saturability at high ion concentrations. This behavior can be quantitatively rationalized by Gouy-Chapman theory, though ion pairing cannot be conclusively ruled out. The finding that transmembrane proteins alter barrier selectivity, favoring polar permeant transport, constitutes an important step toward understanding permeability in biomembranes. PMID- 10667916 TI - Functional characterization of glycosylation-deficient human P-glycoprotein using a vaccinia virus expression system. AB - P-glycoprotein (P-gp), the product of human MDR1 gene, which functions as an ATP dependent drug efflux pump, is N-linked glycosylated at asparagine residues 91, 94, and 99 located within the first extracellular loop. We report here the biochemical characterization of glycosylation-deficient (Gly(-)) P-gp using a vaccinia virus based transient expression system. The staining of HeLa cells expressing Gly(-) P-gp (91, 94, and 99N-->Q), with P-gp specific monoclonal antibodies, MRK-16, UIC2 and 4E3 revealed a 40 to 50% lower cell-surface expression of mutant P-gp compared to the wild-type protein. The transport function of Gly(-) P-gp, assessed using a variety of fluorescent compounds indicated that the substrate specificity of the pump was not affected by the lack of glycosylation. Additional mutants, Gly(-) D (91, 94, 99N-->D) and Gly(-) Delta (91, 94, 99 N deleted) were generated to verify that the reduced cell surface expression, as well as total expression, were not a result of the glutamine substitutions. Gly(-) D and Gly(-) Delta Pgps were also expressed to the same level as the Gly(-) mutant protein. (35)S-Methionine/cysteine pulse-chase studies revealed a reduced incorporation of (35)S-methionine/cysteine in full length Gly( ) P-gp compared to wild-type protein, but the half-life ( approximately 3 hr) of mutant P-gp was essentially unaltered. Since treatment with proteasome inhibitors (MG-132, lactacystin) increased only the intracellular level of nascent, mutant P gp, the decreased incorporation of (35)S-methionine/cysteine in Gly(-) P-gp appears to be due to degradation of improperly folded mutant protein by the proteasome and endoplasmic reticulum-associated proteases. These results demonstrate that the unglycosylated protein, although expressed at lower levels at the cell surface, is functional and suitable for structural studies. PMID- 10667917 TI - Thermal stability of the plasma membrane calcium pump. Quantitative analysis of its dependence on lipid-protein interactions. AB - Thermal stability of plasma membrane Ca(2+) pump was systematically studied in three micellar systems of different composition, and related with the interactions amphiphile-protein measured by fluorescence resonance energy transfer. Thermal denaturation was characterized as an irreversible process that is well described by a first order kinetic with an activation energy of 222 +/- 12 kJ/mol in the range 33-45 degrees C. Upon increasing the mole fraction of phospholipid in the mixed micelles where the Ca(2+) pump was reconstituted, the kinetic coefficient for the inactivation process diminished until it reached a constant value, different for each phospholipid species. We propose a model in which thermal stability of the pump depends on the composition of the amphiphile monolayer directly in contact with the transmembrane protein surface. Application of this model shows that the maximal pump stability is attained when 80% of this surface is covered by phospholipids. This analysis provides an indirect measure of the relative affinity phospholipid/detergent for the hydrophobic transmembrane surface of the protein (K(LD)) showing that those phospholipids with higher affinity provide greater stability to the Ca(2+) pump. We developed a method for directly measure K(LD) by using fluorescence resonance energy transfer from the membrane protein tryptophan residues to a pyrene-labeled phospholipid. K(LD) values obtained by this procedure agree with those obtained from the model, providing a strong evidence to support its validity. PMID- 10667918 TI - Nitric oxide activates or inhibits skeletal muscle ryanodine receptors depending on its concentration, membrane potential and ligand binding. AB - We show that rabbit skeletal RyR channels in lipid bilayers can be activated or inhibited by NO, in a manner that depends on donor concentration, membrane potential and the presence of channel agonists. 10 microm S-nitroso-N-acetyl penicillamine (SNAP) increased RyR activity at -40 mV within 15 sec of addition to the cis chamber, with a 2-fold increase in frequency of channel opening (F(o)). 10 microm SNAP did not alter activity at +40 mV and did not further activate RyRs previously activated by 2 mm cis ATP at +40 or -40 mV. In contrast to the increase in F(o) with 10 microm SNAP, 1 mm SNAP caused a 2-fold reduction in F(o) but a 1.5-fold increase in mean open time (T(o)) at -40 mV in the absence of ATP. 1 mm SNAP or 0.5 mm sodium nitroprusside (SNP) induced approximately 3 fold reductions in F(o) and T(o) at +40 or -40 mV when channels were activated by 2 mm cis ATP or in channels activated by 6.5 microm peptide A at -40 mV (peptide A corresponds to part of the II-III loop of the skeletal dihydropyridine receptor). Both SNAP-induced activation and SNAP/SNP-induced inhibition were reversed by 2 mm dithiothreitol. The results suggest that S-Nitrosylation or oxidation of at least three classes of protein thiols by NO each produced characteristic changes in RyR activity. We propose that, in vivo, initial release of NO activates RyRs, but stronger release increases [NO] and inhibits RyR activity and contraction. PMID- 10667919 TI - Activation and inactivation of mechanosensitive currents in the chick heart. AB - The behavior of MS channels in embryonic chick ventricular myocytes activated by direct mechanical stimulation is strongly affected by inactivation. The amplitude of the current is dependent not only on the amplitude of the stimulus, but also the history of stimulation. The MS current inactivation appears to be composed of at least two contributions: (i) rearrangement of the cortical tension transducing elements and (ii) blocking action of an autocrine agent released from the cell. With discrete mechanical stimuli, the MS current amplitude in the second press of a double press protocol was always smaller than the amplitude of the first MS current. Occasionally, a large MS current occurred when the cell was first stimulated, but subsequently the cell became unresponsive. For a series of stimuli of varying amplitudes, the order in which they were applied to the cell affected the size of the observed MS current for a given stimulus magnitude. When continuous sinusoidal stimulation was applied to the cells, the MS current envelope either reached a steady state, or inactivated. With sinusoidal stimulation, the MS response could be enhanced or restored by simple perfusion of fluid across the cell. This suggests that mechanical stimulation of the cells produces an autocrine inhibitor of MS channels as well as resulting in cortical rearrangement. PMID- 10667920 TI - Whole-cell mechanosensitive currents in rat ventricular myocytes activated by direct stimulation. AB - Mechanosensitive channels may have a significant role in the development of cardiac arrhythmia following infarction, but the data on mechanical responses at the cellular level are limited. Mechanosensitivity is a ubiquitous property of cells, and although the structure of bacteriological mechanosensitive ion channels is becoming known by cloning, the structure and force transduction pathway in eukaryotes remains elusive. Isolated adult rat ventricular myocytes were voltage clamped and stimulated with a mechanical probe. The probe was set in sinusoidal motion (either in, or normal to, the plane of the cell membrane), and then slowly lowered onto the cell. The sinusoidal frequency was held constant at 1 Hz but the stimulation amplitude was increased and the probe gradually lowered until a mechanically sensitive whole cell current was seen, which usually followed several minutes of stimulation. The whole cell mechanosensitive current in rat cells had two components: (i) a brief large inward current spike current; (ii) a more sustained smaller inward current. The presence of the initial sharp inward current suggests that some structure within the cell either relaxes or is broken, exposing the mechanosensitive element(s) to stress. Metabolic changes induced by continued stress prior to the mechanosensitive response may weaken the elements that break producing the spike, or simple stress-induced fracture of the cytoskeleton itself may occur. PMID- 10667922 TI - Evaluation of Co-solvents with supercritical fluid extraction of atrazine from soil. AB - Supercritical fluid extraction (SFE) with CO(2) has been successfully applied to herbicide extractions from soil. The objectives of this work were to compare extraction efficiency of atrazine from soil using different types and quantities of co-solvent modifiers under a specified set of SFE instrument conditions and to determine the ruggedness of an optimized extraction program and co-solvent on several soils with varying characteristics. The effect of 18 co-solvents on atrazine extraction from Lufkin fine sandy loam was determined using a completely randomized design with six replications. Extractions of Lufkin soil using the more nonpolar co-solvents had recovery similar to extractions where no co-solvent was added. The co-solvents that showed high extraction efficiency, low incidences of restrictor plugging, and ease of cleaning extraction cells were acetone, acetone:water mixtures (with and without 1% triethylamine), and acetonitrile. The addition of 1% triethylamine (TEA) did not increase recovery significantly. The 9:1 acetone:water mixture with 1% TEA was used for the soil comparison because of the high atrazine recovery and low water content. No differences in atrazine recovery were detected between extractions of the four representative soils when the same extraction conditions were employed. No cleanup steps were included in the procedure, yet adequate chromatography results were obtained suggesting some selectivity for this procedure. These data indicate that SFE with optimized conditions and appropriate co-solvents is a relatively robust method that can effectively be used in soil extractions of atrazine. PMID- 10667923 TI - Fungal biomass in saltmarsh grass blades at two contaminated sites. AB - Ascomycetous fungi are the principal drivers of the decomposition of shoots of smooth cordgrass (Spartina alterniflora). Shoots of smooth cordgrass move into the saltmarsh food web via the decomposition system. Therefore, influences on saltmarsh ascomycetes by pollutants of saltmarshes could have far-reaching impacts. Earlier examination of impacts of severe contamination of a Georgia saltmarsh by mercury and polychlorinated biphenyls (PCBs) revealed little or no influence of the toxicants on living standing crops or sexual productivities of cordgrass ascomycetes. Extension of the examination of saltmarsh-ascomycete response to sites containing other toxic pollutants (the chlorinated organocyclic insecticide toxaphene; chromium, copper, and lead; and polycyclic aromatic hydrocarbons [PAHs]) has shown that none of the additional toxicants engendered saltmarsh-fungal responses in the form of reduced living standing crops or sexual productivities. Thus the ascomycetes of the cordgrass-decay system appear to be as resistant to anthropogenic-pollutant poisoning as smooth cordgrass itself. Unless the fungal and plant resistance mechanisms involve degradation of the toxicants, this may imply that saltmarshes are especially dangerous as receiving sites for toxic waste because they may have the potential to readily move toxicants into the food web. PMID- 10667924 TI - Phototoxic evaluation of marine sediments collected from a PAH-contaminated site. AB - The phototoxicity potential of PAH-contaminated field sediment was evaluated and compared to standard sediment toxicity test results. Marine sediments were collected from 30 sites along a presumed PAH sediment pollution gradient in Elliot Bay, WA. Standard 10-day acute and 28-day chronic sediment toxicity tests were conducted with the infaunal amphipods Rhepoxynius abronius and Leptocheirus plumulosus using mortality and the ability to rebury as endpoints. The survivors of these tests were then subjected to 1-h exposures to UV radiation with mortality and reburial again determined. The most highly toxic sediments identified in these experiments were evaluated further for toxicity and phototoxicity by serially diluting them with uncontaminated sediment and repeating the toxicity tests. Standard 10-day toxicity test results indicated that over 70% of the sites sampled in Elliot Bay exhibited measurable toxicity with nine sites being highly toxic to both species of amphipods. Results of standard 28-day chronic sediment toxicity tests were similar. In contrast, almost all of the sites were found to be highly phototoxic. Results indicated that exposure to UV increased toxicity five- to eightfold. This suggests that standard toxicity tests underestimate the potential ecological risk of PAH-contaminated sediments in animals exposed to sunlight. However, only when PAH contamination was between 0.05 and 1.0 toxic units would conducting a phototoxicity evaluation add information to that gained from conducting a standard sediment toxicity test alone. PMID- 10667925 TI - Factors controlling the bioaccumulation of mercury, methylmercury, arsenic, selenium, and cadmium by freshwater invertebrates and fish. AB - Concentrations of mercury (Hg), methylmercury (MMHg), arsenic (As), selenium (Se), and cadmium (Cd) were measured in atmospheric deposition, stream water, and biota in two streams in western Maryland. Overall, concentrations were slightly higher in the water of the lower pH Herrington Creek tributary (HRCT). Bioaccumulation factors were also higher for HRCT compared to Blacklick Run (BLK). MMHg concentrations in biota increased with trophic level and essentially all the Hg was as MMHg in predatory insects and insectivorous/carnivorous fish. Thus, the overall trophic status of the organism was indicated by the %MMHg in its tissues. Levels of As, Se, Cd, and Hg, however, decreased with increasing trophic level. Adsorption of As to the exoskeleton of invertebrates appears to be an important accumulation mechanism. MMHg was distributed evenly throughout crayfish and fish organs, whereas As, Se, Cd, and Hg were found in higher concentrations in detoxifying organs. Concentrations in biota in this study were somewhat elevated compared to other rural sites, but were less than those of point source-contaminated sites. Overall, as atmospheric inputs to the two watersheds were similar, the results of this study show the importance of water chemistry in determining the bioaccumulation of the metals and metalloids into insects. Subsequent transfer to higher trophic levels is related to both the ability of the organisms to depurate and the mode of accumulation, either directly from water or from food. PMID- 10667926 TI - Toxicity of manganese to Ceriodaphnia dubia and Hyalella azteca. AB - Manganese is a toxic element frequently overlooked when assessing toxicity of effluents, sediments, and pore waters. Manganese can be present at toxic levels in anoxic solutions due to increased solubility under chemically reducing conditions, and it can remain at those levels for days in aerated test waters due to slow precipitation kinetics. Ceriodaphnia dubia and Hyalella azteca are freshwater organisms often used for toxicity testing and recommended for assessments of effluents and pore waters. Lethal and reproductive-inhibition concentrations of Mn were determined for C. dubia in acute 48-h tests and chronic three-brood tests using animals <24 h old and between 24 and 48 h old. Sensitivity of H. azteca to Mn was determined with 7-day-old animals in acute 96 h tests. Tests were run at three levels of water hardness to assess the amelioratory effect, which was often significant. Manganese concentrations were measured analytically at test initiation and after 96 h for calculation of toxicity and determination of Mn precipitation during the tests. Minimal amounts of Mn (30 mU/l) in preparation for a whole-body scintigraphy. The whole-body scintigraphy showed tumour tissue or metastases in 16 patients (group C). Owing to an inconclusive whole-body scintigraphy, 17 patients were classified as being in remission (group B). After levothyroxine substitution 14 patients of group B were measured in euthyroidism too (group D). Besides these sub-groups, there was a control group made up of 23 healthy volunteers (group A). Group B showed significantly higher PE + SM and PC concentrations than group C (0.59 +/- 0.02 mmol/l PE + SM in B vs 0.48 +/- 0.02 mmol/l in C; 2.1 +/- 0.1 mmol/l PC in B vs 1.6 +/- 0.1 mmol/l in C). In comparison with group D higher concentrations of the phospholipids PE + SM and PC as well as PI were found in group B (0.59 +/- 0.02 mmol/l PE + SM in B vs 0.48 +/- 0.03 mmol/l in D; 0.074 +/ 0.005 mmol/l PI in B vs 0.046 +/- 0.004 mmol/l in D; 2.1 +/- 0.1 mmol/l PC in B vs 1.6 +/- 0.1 mmol/l in D). The data indicate that under the condition of hypothyroidism only patients in remission (group B) show significantly increased phospholipid concentrations, whereas the values in patients with remaining tumour tissue (group C) do not differ from those of the reference groups A and D. This finding is interpreted as an interference between the hormonal status and the systemic effects of cancer. PMID- 10668050 TI - 31P MRS measurement of mitochondrial function in skeletal muscle: reliability, force-level sensitivity and relation to whole body maximal oxygen uptake. AB - The reliability, relation to whole-body maximal oxygen uptake (VO(2max)), and force-level sensitivity of (31)P MRS markers of mitochondrial function were studied in 39 normal-weight women. Following 90 s isometric plantar-flexion exercises at 45, 70 and 100% of maximum voluntary contraction, skeletal muscle mitochondrial function was determined from the phosphocreatine recovery time constant (TC(PCr)), the ADP recovery time constant (TC(ADP)), and the rate of change in PCr during the first 14 s of recovery (OxPhos). VO(2max) was measured on a treadmill. Test-retest measurements were obtained in a subset of seven women. Overall, TC(PCr), TC(ADP) and OxPhos were reproducible for all exercises (coefficients of variation = 2.3-19.3%). With increasing force-level, TC(PCr) was prolonged (29.0 +/- 8.2, 31.9 +/- 9.0 and 35.4 +/- 9.5 s), OxPhos was increased (0.159 +/- 0.081, 0.247 +/- 0.090 and 0.310 +/- 0.114), and TC(ADP) was shortened (22.4 +/- 7.9, 21.3 +/- 6.2, and 19.5 +/- 6.7; p < 0.01). All MRS markers of mitochondrial function were correlated with VO(2max) (r = 0.41-0.72; p < 0.05). These results suggest that measurements of TC(PCr), TC(ADP) and OxPhos yield reproducible results that correlate with whole-body VO(2max) and that vary in force-level sensitivity. PMID- 10668051 TI - On the reliability of quantitative clinical magnetic resonance spectroscopy of the human brain. AB - The reliability of a single-voxel, localized proton magnetic resonance spectroscopy protocol suitable for clinical studies was investigated by means of in vitro, single-subject in vivo and multi-subject in vivo examinations of healthy adults aged from 19 to 67 years. The study was performed at 1.5 T using a standard quadrature head coil and a single voxel PRESS sequence (in vitro TR/TE = 1500/30 ms, in vivo TR/TE=2000/35 ms). Eighty-four in vitro and 30 single-subject examinations were statistically evaluated after quantification, including the calculation of the coefficients of variations (CV) for choline (Cho), creatine (Cr), myo-inositol (mI), lactate (Lac), N-acetyl-aspartate (NAA) and unresolved glutamine, glutamate and GABA (Glx). The CVs for absolute concentrations of the main metabolites Cho, Cr and NAA, ranged from 3.3% (3.8) to 4.0% (6.4%) (the in vivo results are given in brackets). Multi-subject CVs of absolute concentrations for Cho, Cr and NAA ranged from 7.6% to 15.0%. CVs of relative in vivo concentrations were found to be higher than CVs of absolute concentrations. Due to the better reproducibility of intra-individual absolute in vivo concentrations, cross-over studies using institutional units are recommended. PMID- 10668052 TI - Simultaneous detection of changes in perfusion and BOLD contrast. AB - A new functional magnetic resonance imaging (fMRI) technique for simultaneous detection (SIDE) of changes in perfusion and blood oxygenation level dependent (BOLD) contrast is described. Perfusion contrast is generated by using magnetically labeled endogenous water proton spins as a freely diffusible tracer. A single slice-selective inversion pulse is combined with dual echo echo-planar imaging to generate a spin-echo (SE) image sensitive to changes in perfusion and a gradient-echo (GE) image sensitive to changes in both perfusion and BOLD contrast. The SIDE technique was applied to detect functional changes induced by a visual search task. A theoretical analysis is provided to calculate quantitative maps of changes in cerebral blood flow (DeltaCBF) and effective transverse relaxation time (DeltaT(2)*) from the corresponding signal changes in the SE and GE images. Since SE an GE images are generated from the same longitudinal magnetization, no errors due to spatial or temporal mismatch can arise in the quantification of DeltaCBF and DeltaT(2)*. PMID- 10668053 TI - Regional brain activation by bicuculline visualized by functional magnetic resonance imaging. Time-resolved assessment of bicuculline-induced changes in local cerebral blood volume using an intravascular contrast agent. AB - Functional magnetic resonance imaging (fMRI) has been applied to study rat focal brain activation induced by intravenous administration of the GABA(A) antagonist bicuculline. Using magnetite nanoparticles as a blood pool contrast agent, local changes in cerebral blood volume (CBV) were assessed with high temporal (10 s) and spatial (0.35 x 0.6 mm(2)) resolutions. Upon infusion of the bicuculline region-specific increases in CBV have been observed, suggesting CBV to reflect brain activity. During the first 2 min, the signal increases were predominant in the cortex, followed by increases in other brain areas, such as the caudate putamen, thalamus and cerebellum. Ten minutes after the start of infusion, a dominant response was observed in the thalamus, while in the caudate putamen a biphasic response pattern was seen. The magnitude of the signal responses in all brain regions was dependent on the dose of bicuculline and, in general, matched the known distribution of GABA(A) binding sites. This study suggests that pharmacological fMRI, displaying brain function at the highly specific level of drug-receptor interaction, should foster our understanding of normal and pathological brain function. PMID- 10668054 TI - Child survivors of parental death from cancer or suicide: depressive and behavioral outcomes. AB - Depressive symptoms, social competence, and behavior problems of prepubescent children bereaved within 18 months of parental death from cancer (57 families, 64 children) or suicide (11 families, 16 children) were compared. Most children reported normative levels of depressive symptoms. Children whose parents died from suicide, compared with those whose parents died from cancer, reported significantly more depressive symptoms, involving negative mood, interpersonal problems, ineffectiveness, and anhedonia. Parental reports of children's competence and behavior were similar to a normative sample of children and did not differ between the children bereaved by parental cancer or suicide. Additional research should focus on other factors, such as family psychopathology, stresses, and impact of stigma, which may influence the course of bereaved children. PMID- 10668055 TI - Promoting psychological well-being in the face of serious illness: when theory, research and practice inform each other. AB - This article describes the interplay among theory, research and practice regarding the maintenance of psychological well-being during serious illness. The ideas emerged from two independent lines of work, one that evolved through clinical practice within the medical model, the other that evolved through theory and field research within a behavioral science model. Each of these lines of work independently points to the importance of focusing on psychological well-being and the coping processes that support it, as a complement to the traditional focus in both the medical and behavioral sciences on psychiatric symptoms. This article describes a theoretical framework for the discussion of psychological well-being during serious illness. Then, this framework is used to define variables that research indicates contribute specifically to psychological well being during serious illness, and finally, based on theory and research, a therapeutic program is described for patients with serious illness. The goal of this paper is to encourage researchers and clinicians to give as much attention to the development and maintenance of psychological well-being in the face of serious illness as they do to the etiology and treatment of psychiatric symptoms. PMID- 10668056 TI - Negative and positive influences of social support on depression in patients with head and neck cancer: a prospective study. AB - Patients with head and neck cancer have to cope not only with a life threatening diagnosis, but also with an altered facial appearance and the loss or impairment of important functions as a result of treatment. As a consequence they are prone to psychosocial problems. Social support might influence their ability to adapt to the illness and its treatment. The aim of this prospective study is to examine the influence of different aspects of social support on the depressive symptomatology in head and neck cancer patients treated with surgery and/or radiotherapy. Patients completed a questionnaire relating to available and received support, the extent of the social network, depressive symptoms, and general health complaints before and 6 months after treatment. Received support was found to be associated with more depressive symptomatology at baseline and available support led to less depressive symptomatology. The relationship between social support and depressive symptoms was especially apparent in patients with few general health complaints. Whereas the availability of support seemed to be beneficial regardless of the situation, the effect of received support was equivocal. The provision of support should be tailored to the needs of the individual patient. PMID- 10668057 TI - Cisplatin-based therapy: a neurological and neuropsychological review. AB - The present paper reviews research in the area of the broad-spectrum chemotherapeutic agent cisplatin (cis-diamminedichloro-platinum II) and examines the implications for clinical neuropsychology arising from the neurological disruption associated with cisplatin-based therapy. The paper begins with a brief review of cisplatin treatment in terms other than survival alone, and examines the side-effects and the potential central nervous system (CNS) dysfunction in terms of neurological symptoms and concomitant implications for neuropsychology. Two main implications for clinical neuropsychology arising from cisplatin therapy are identified. First, cisplatin therapy impacts upon the psychological well being of the patient, particularly during and in the months following treatment. It is suggested that during this time, a primary role for neuropsychology is to focus upon the monitoring and the active enhancement of the patient's social, psychological and spiritual resources. Second, with regard to neurocognitive changes, the review suggests that (1) neurocognitive assessment may not yield stable results within 8 months following treatment and (2) while perceptual, memory, attentional and executive dysfunction may be predicted following cisplatin treatment, little systematic research has been carried out to investigate such a possibility. Future research might profitably address this issue and also specifically examine the effects of low dosage cisplatin-based therapy and the effects of recently developed neuroprotective agents. Finally, there is some evidence to suggest that women may be more susceptible to neurotoxicity during cisplatin therapy, but no gender-related cognitive effects are reported in the cisplatin literature. Future research could usefully investigate gender differences in association with cisplatin chemotherapy. PMID- 10668058 TI - Using beliefs and magical thinking to fight cancer distress-a case study. AB - This case relates to the way in which a young patient developed serious difficulties in coping with her life in the years following a successful bone marrow transplant. By means of an illustrative metaphor, she revealed her existential position and the way in which she attempted to deal with her anxiety. Being diseased implied that life's order was replaced by disorder and a loss of basic trust. She tried to re-establish order by establishing beliefs that attributed specific regularities to life, and to influence the risk of recurrence by living according to these beliefs. Unfortunately, this meant that she had to tread a very thin line over a course mined with anxiety and eventually, she became a prisoner of her own creation. The author claims that we can learn from this case, as it clearly illustrates psychological dimensions commonly seen in cancer patients: the way anxiety is related to disorder and the way patients try to regain control of their lives through constructing belief-systems. The case also features a discussion of how we, as clinicians, may be able to help these patients. PMID- 10668059 TI - A pilot study of interpersonal psychotherapy by telephone with cancer patients and their partners. AB - A single-arm pilot study explored the feasibility of adapting in Interpersonal Psychotherapy (IPT) by telephone to reduce psychological distress and to enhance coping during cancer treatment. Therapy focuses on role transitions, interpersonal conflicts, and grief precipitated by cancer. Breast cancer patients receiving high-dose chemotherapy received weekly sessions with a psychologist throughout chemotherapy and for 1 month afterwards. Patients could invite one 'partner' to receive individual telephone IPT. Psychosocial functioning was assessed using standardized measures at study entry, after chemotherapy, and following telephone IPT. Accrual and participation supplied evidence of feasibility: 14 patients and 10 partners were recruited, 82.5% of those eligible. Patients had a mean of 16 sessions; partners had a mean of 11. Participants rated their satisfaction with the program between 'good' and 'excellent'. A test of the efficacy of telephone IPT requires a larger, randomized trial. In order to standardize the intervention, a treatment manual was developed. This study indicated the importance of outreach to family members as well as to cancer patients, intensive patient education about oncology treatment and the medical care setting, and psychosocial services that continue after cancer treatment has been completed. PMID- 10668060 TI - The psychosocial experience of women treated for breast cancer by high-dose chemotherapy supported by autologous stem cell transplant: a qualitative analysis of support groups. AB - Autologous bone marrow transplantation (AuBMT) is probably among the most aggressive of physical treatments endured by cancer patients. High-risk breast cancer patients who choose this therapy face prolonged, agonizing and life threatening interventions that are no less arduous than confronting the malignant disease itself. The study, which aimed to broadening our understanding of the psychosocial impact and the implications of AuBMT, presents a protocol analysis of group support intervention in 45 recipients (eight to ten women in five groups). The sessions were held at the Transplant Department at the Chaim Sheba Medical Center. The contribution of group support to the healing process was examined. The findings show that recovery was affected by a wide range of psychosocial factors, specifically highlighting the impact of transplantation and survival on five domains, viz. physical, psychological/emotional, vocational, social and family/spousal intimacy. Illness and treatment management is also discussed. The support generated by the group, both individually and collectively, was found to contribute significantly to the spectrum of resources available to the participants. PMID- 10668061 TI - Stability and change in posttraumatic stress disorder symptoms following breast cancer treatment: a 1-year follow-up. AB - While some recent research has examined the prevalence and severity of posttraumatic stress disorder (PTSD)-like symptoms following cancer treatment, no research has examined temporal change or stability in these symptoms in cancer survivors. Female breast cancer survivors (n=46) participated in an initial telephone interview and a follow-up interview 12 months later. PTSD symptoms associated with breast cancer were assessed using the PTSD Checklist-Civilian version (PCLC). In general, PTSD symptoms in this population did not diminish over time. While group analyses indicated that PCLC-total and subscale scores were stable across the two assessments, analyses of PCLC scores indicated that many patients exhibited fairly large (>0.5 S.D.) increases and/or decreases in PCLC-total or subscale scores. Some evidence suggested that decreases in PCLC scores between the two study assessments were associated with greater social support and experience of fewer traumatic stressors prior to breast cancer diagnosis. Most significantly, the research suggested that women with greater PTSD symptoms at the initial interview were less likely to participate in the follow-up interview. Implications of this for research and clinical management of PTSD in this population are discussed. PMID- 10668062 TI - Development of a scale to measure psychosocial concerns of Mexican women with advanced cancer. AB - This descriptive study was undertaken to explore the perceptions and attitudes of 143 women (mean age+/-51) with advanced cancer. Outcome measures were the Perceptions and Attitudes Scale (PAS), a 65-item instrument validated as part of the study, and the Karnofsky Performance Status Scale. Internal consistency (Cronbach alpha) and factor analytic techniques (principal component and orthogonal rotation) were performed. The scale has an internal consistency of 0. 79. Factor analysis revealed six factors which explained 39.6% of the variance (1) loneliness; (2) family and social sphere; (3) limitations and dependence conditions; (4) perceptions about their lives; 5) perceptions about their health and pain; and (6) perceptions of pain and anguish. These results point out the necessity of a multivariate approach to women with advanced cancer. PMID- 10668063 TI - Remission of chemotherapy-induced emesis with concurrent olanzapine treatment: a case report. PMID- 10668064 TI - S. P. Cotton, E. G. Levine, C. M. Fitzpatrick, K. H. Dold and E. Targ, 'Exploring the relationships among spiritual well-being, quality of life, and psychological adjustment in women with breast cancer'. psycho-oncology 8(5) 1999, 429-438 AB - The original article to which this Erratum refers was published in Psycho-Ocology 8(5) 1999, 429-438. PMID- 10668065 TI - [Seroprevalence of helicobacter pylori infection in the republic of Argentina: influence of age, sex, socioeconomic level, geographical area, and health infrastructure. Multicenter study by the Club Argentino del Estomago y Duodeno]. AB - Helicobacter pylori (Hp) infection affects almost half of the world population, it is almost a pandemia, and has been associated to poverty in underdeveloped countries. The Club Argentino del Estomago y Duodeno decided to fulfill the lack of information upon this subject in Argentina designing a seroprevalence, multicentric, prospective study performed in voluntary adults donors in blood banks and in children seen during normal growth controls. Seven hundred and nineteen individuals were evaluated, 645 of them were included: 178 children (age 0-18 years) and 467 adults. In all cases a serological IgG Hp test (Flex-Pack Abbott) was performed and an epidemiological questionnaire was completed by a physician. General prevalence of Hp infection was 44.8% of individuals. In the paediatric population prevalence was 15.7% and in adults 55.9%. The highest prevalence was observed in the fifth decade: 64%. In concordance with other similar studies carried out in different countries, we may conclude that the risk of acquisition of Hp infection is directly related to age, area of residence, social-economical status, sanitary facilities, and educational level reached. Even though the prevalence of Hp infection in Argentina is intermediate between highly developed and underdeveloped countries, the number of people infected is very high and the incidence of Hp-associated pathologies in the future represents a formidable task for gastroenterologists and sanitary authorities. PMID- 10668066 TI - [Moderating action of celiac block in experimental pancreatitis in the dog]. AB - BACKGROUND: Pancreatic ischemia seems to be responsible of transition from edematous to hemorrhagic forms in acute pancreatitis. (AP) Sympathetic system vasoconstriction, through celiac plexus play an important role in the pathogenesis of acute pancreatitis. OBJECTIVE: Determinate the effects of anesthetic celiac blockade in an experimental model of AP. METHODS: Distal pancreatectomy and intraductal injection of autologous bile in 28 mongrel dogs. Blockade of celiac plexus with bupivacaine in the experimental group B. Anatomopathologic examination after 72 hours. RESULTS: Experimental group B developed milder forms of AP, while the control group A developed severe forms. CONCLUSIONS: Results suggest that celiac plexus blockade with bupivacaine may prevent the development of necrohemorrhagic forms of PA in a canine model. PMID- 10668067 TI - [Intraepithelial enteroendocrine cells in cecum and appendix from ovoalbumin sensitized rabbits]. AB - Intraepithelial enteroendocrine cells (IEC) produce peptides which influence motility, secretion and absorption of nutrients. Recently the role of these cells in the immune mucosal system is under study. The aim of the present study was to evaluate the modifications in number of IEC in cecum and appendix from Ovalbumin (OVA) sensitized rabbits. Twenty adult New Zealand rabbits were separated in two groups: Group 1 (G1 = 10) not sensitized normal control. Group 2 (G2 = 10) were sensitized twice intraperitoneally with 70 mg OVA and 30 mg ALUM/ml (aluminium hydroxide). Anti OVA specific IgE was evaluated by means of PCA test (passive cutaneous anaphylaxis). Samples form cecum and appendix were fixed in buffered formaldehyde 10%, paraffin embedded and stained with anti-Chromogranin A for neuroendocrine cells. 400 high power fields were counted in each animal, referred as IEC/100 enterocytes. In cecum surface epithelium and crypt were considered. Surface epithelium, deep crypts and superficial crypts were evaluated in appendix. Results showed in cecum in G1:1,6 IEC/100 enterocytes in surface epithelium and 3/100 in crypts; G2 6 IEC/100 in surface epithelium and 12/100 in crypts. The difference between G1 and G2 was statistically significant (p < 0.05). In appendix surface epithelium from G1 showed 5.2/100 while G2 5.4/100. Superficial crypts 8.5 (G1) and 11.3 (G2) (p < 0.05) and deep crypts 4.9 (G1) and 8.5 (G2) (p < 0.01). The results showed that OVA-sensitized animals presented increment in the number of IEC in surface epithelium and crypts which may indicate a relationship between these cells and rabbit mucosal immune system. PMID- 10668068 TI - [Cancer of the esophagus and stomach: 3-year evaluation]. AB - Esophagus cancer has a very bad pathological prognosis. Risk factors considered are: smoking consumption and deficiency of vitamins A and C. The mortality rates of cancer of the stomach vary notably according to geographic region. Factors such as genetics, races, smoking, socioeconomic conditions are some of stomach cancer development. 646 symptomatic patients were studied in the gastroenterology unit at. J.M. Cullen hospital. Histopathologically, 22 (3.3%) cancer of the esophagus and 13 (2%) cancers of stomach were detected. All esophagus cancers were squamous cells; 82% were males and 18% females. 50% were located on the middle third zone. 92.3% of stomach cancers were adenocarcinoma; 83% were males and 17% females. A 50.8% were located in corporo-fundic zone. All the cancers both of the esophagus as well as of the stomach, were in the advanced phase. The cancer of esophagus has appeared most frequently among cancers of the tract in our hospital, with significant difference among province and national registers. PMID- 10668069 TI - [Vascular ectasias of the gastric antrum as the cause of chronic digestive hemorrhage. Presentation of a case]. AB - Digestive hemorrhages are one of the syndromes, that frequently attracts the attention of the gastroenterologist-endoscopist. Sometimes it's not evident during upper endoscopy study and course with a contrast radiological scan. In the last years a vascular pathology has been seen which is responsible of high digestive hemorrhages. It is manifested by melaena and is originated by vascular ectasias, which are vascular dilatations present in the stomach or right colon during various accompanying pathologies like hepatic cirrhosis with portal hypertension, cardiac valvulopaties, etc. Here we present a 61 year old woman with a history of chronic hepatopathy of cirrhotic type and imprecise etiology (diagnosed since 1983). Approximately two years ago (1996-1997) she has been presenting digestive hemorrhages and she has been transfused in different occasions because she had severe anemia. Diffuse vascular ectasias in the distal region of the antrum and in part of the gastric body were observed during duodenoscopy and colonoscopy with signs of active hemorrhage and similar non bleeding lesions in the colon. Pyruvic transaminase was normal; HBV and HCV markers were -negative. Ultrasound analysis was normal. The diagnosis at the discharge from Hospital was Diffuse Vascular Ectasias of the Gastric Antrum and part of the gastric body, caecum and right colon, secondary to hepatic cirrhosis. She was admitted again at the hospital because of new upper digestive hemorrhages and surgery was indicated. Gastric resection was performed with promising results. This case is analyzed and the pathology is results. PMID- 10668070 TI - [Computed tomography-guided treatment of infected acute pancreatitis]. AB - We report a case of a 60 years old man admitted for infected acute pancreatitis and duodenal fistula with mediastinal extension at admission, successfully treated in a 40-days period with CT-guided percutaneous catheter drainage (Malecot 14F) inserted with Seldinger method, in spite of a colic fistula development during the treatment. PMID- 10668071 TI - [Cancer of the esophagus and the stomach: should we become resigned or investigate?]. PMID- 10668072 TI - Molecular biology diagnosis of hereditary non-polyposis colorectal cancer (HNPCC) PMID- 10668073 TI - [Gastroesophageal reflux. To operate or to wait?]. PMID- 10668074 TI - [Suicides by police officials in North Rhine-Westphalia. An evaluation of 58 suicides between 1992 and 1998]. PMID- 10668075 TI - [Value of fingerprints for determining identity. Simultaneously a contribution on significance for reconstruction of the behavior of a perpetrator based on the evidence--trends over time]. PMID- 10668076 TI - [Death caused by neglect?]. AB - Over a period of several months a 23-year-old bedridden woman suffered from increasing cachexia with numerous decubital ulcera in the parental house. The parents induced the hospitalization not before the patient reached the agonal phase. The reported case was especially spectacular because the left arm had fallen off and lay beside the woman showing maggot infestation, mummification and skeletonization. Against the parents a preliminary investigation for manslaughter by negligence was instituted. This case report describes the unusual course of disease and the striking familiar situation. PMID- 10668077 TI - [Appearance of injuries caused by machetes and unusually large knives]. AB - Machetes (Spanish-American matchets) are sharp, long knives with a broad blade having a slightly curved edge and a thick back. They are used for clearing paths in rough, densely wooded areas. Just like axes and swords they are suitable for causing not only soft tissue wounds, but also deep slashes in the underlying bone. In military conflicts (especially in Central Africa) they are often used as short-range weapons, whereas it is in the nature of things that they are rarely used in Europe. On the basis of 5 cases from the Freiburg material fatal and survived injuries caused by machetes and similarly large knives are discussed. In 2 cases long chop injuries were inflicted on the head causing sharp-edged transections of the bony skullcap; in one case in which the victim survived a broken off part of the blade lodged in the right os parietale. In another 2 cases complete and subtotal decapitation respectively occurred in the victims. As the instrument can be used both for cutting and stabbing there were not only long cutting injuries, but also gaping stab wounds severing the skin over a length of several centimetres; due to the great length of the blade through-and-through wounds were also seen on the trunk. Massive blows with the thick back of the blade caused streak-like bruising. PMID- 10668078 TI - [Examples for use of the STR systems in stain assessment with special reference to Y chromosome systems]. AB - The authors describe 3 cases where Y-chromosomal systems were used for typing the biological traces. In the first case, a murder, for the major amount of cell material found on a dish towel (and analysed two years after the crime) female persons were excluded for the system amelogenin and Y-chromosomal systems. A brother of the victim could not be excluded for autosomal STR-systems. Upon confrontation with the results of the DNA-analysis (among other things), this brother confessed the murder of his sister some days later. He was found guilty by the court. In the second case described, a rape of two girls, many traces were analysed parallely with Y-chromosomal and autosomal PCR-systems. The objects where male DNA matching the suspect were found (a paper tissue, a sweat shirt and the knickers of the girls), also showed small amounts of alleles matching with the suspect for autosomal systems, while the major part in these systems was from the girls. The suspect was sentenced to many years imprisonment. In the third case, a possible rape of a young woman, a stained microscope slide of a vaginal swab had to be examined. Microscopically a few sperm heads could be seen in a surplus of leucocytes. The male proportion could be analysed only in the Y chromosomal systems, not in the autosomal ones. For the frequency calculation of the Y-chromosomal allele combination the haplotype data bank of the Institute for Legal Medicine of the Humboldt university in Berlin was indispensable. PMID- 10668079 TI - [Mitochondrial disfunction and perinatal exposure to antiretroviral nucleoside analogues]. PMID- 10668080 TI - [Laryngeal foreign bodies: management in children in Senegal]. AB - Inhalation of foreign bodies is a frequent accident in children. It remains severe in the case of laryngeal foreign bodies. PATIENTS AND METHODS: Retrospectively, for a 16-year period, 65 laryngeal foreign bodies have been treated (44.8%), among 145 cases of airway foreign bodies, in the ENT department of Dakar University hospital. Etiological, clinical and therapeutic aspects were reviewed. RESULTS: Average age was 36 months, with a sex-ratio of 2.42 in favour of males. The time lag (time between the accident and admission to the department) was particularly long; 73.33% of the children were admitted more than 24 hours after the event. Eighty-three percent of the patients presented greater or lesser laryngeal dyspnea. Tracheostomy was performed in 55.4% of the patients. Average duration for abiation of the canula was ten days. Three cases of death were recorded (4.16%). DISCUSSION: The frequency of 44.8% for laryngeal localization of foreign bodies appears to be the highest in the literature. If the appropriate treatment for foreign bodies in the respiratory tract is endoscopic removal, the tracheostomy nevertheless occupies a central place in the management of the disease. This procedure may be recommended to all ENT specialists working in similar conditions. In spite of its inherent complications, tracheostomy allows reduction of mortality in relation to laryngeal foreign bodies. Improvement of prognosis requires prevention based on widespread public information and improving technical infrastructures. PMID- 10668081 TI - [Sickle cell disease in children in Dakar, Senegal]. AB - AIM OF THE STUDY: To determine the socioeconomic, clinical and biological aspects of sickle cell disease (SCD) in Senegalese children and adolescents, we retrospectively analysed all records of follow-up attending patients in the Albert Royer Children Hospital of Dakar (Senegal). RESULTS: Homozygous sickle cell (SS) was the most frequent genotype (307 cases). Sickle cell hemoglobin C (13 cases) and sickle cell beta-thalassemia (three cases) were uncommon. Patients were aged from five months to 22 years (mean age: eight years). Most of them came from poor families. The mean number of children was five in patients' families, with at least two cases of SCD in 60% of them. Immunization against hepatitis B virus (10.2%), Haemophilus influenzae b (8.4%), Salmonella (8.7%) and Streptococcus pneumoniae (21.4%) was insufficiently performed, because of its relatively high cost. Only 30% of the patients had received a blood transfusion. Painful crises occurred less than three times a year in 74% of the cases. Complications such as acute chest syndrome (1%), stroke (1%), cholelithiasis (9%), meningitis (0.4%), septicemia (2%) and osteomyelitis (6%) were rare. Mean steady state hemoglobin (Hb) and hemoglobin F(HbF) levels were 8.27 +/- 1.36 g/dL and 6.8 +/- 5.9% respectively among SS patients. No correlations were found neither between Hb and HbF nor between these parameters and the frequency of complications. Eleven patients (1.1% per year of follow-up) died, and infection was the main cause of death (73%). CONCLUSION: In comparison with published data, SCD seems to have mild severity in Senegalese children and adolescents in spite of poor follow-up conditions. In addition to genetic factors, environmental factors might have an important role in disease tolerance. PMID- 10668082 TI - [Adolescents and ambulatory care: results from a national survey of young people 15 to 20 years of age in Switzerland]. AB - In industrialized countries, statistics on health services exhibit a low level of health care use by adolescents, despite the fact that their needs have been widely described. OBJECTIVES: To assess ambulatory health care use by 15-20-year old teenagers in Switzerland. METHOD: Nine thousand, two hundred and sixty-eight adolescents responded to the self-administered questionnaire distributed in secondary schools and vocational classes for the Swiss Adolescent Health Survey. Questions about visits to general practitioners, specialists and gynecologists, reasons for visit, the availability of a regular health care provider and a confidential health care resource were analysed. RESULTS: Within the previous 12 months, 87.6% of the girls and 75.3% of the boys reported having seen a physician. General practitioners were visited more frequently than specialists. The contact with a specialist was the only one to be related to socio-demographic variables: a lower proportion of reported visits to a specialist was related to apprenticeship, low educational status of parents or rural living area. Thirty nine percent of the girls reported having seen a gynecologist during the previous 12 months. Two adolescents out of three reported having a personal doctor, and one out of two declared being aware of a confidential health care resource. Girls reported a larger number of reasons for visits than boys: chronic conditions, fatigue, headache and depressive symptoms were the most often cited in a list of ten reasons. Among the subjects who declared a health concern (sleep disturbances, eating disorders, depressive symptoms, smoking or alcohol-related problems) and a need for help, less than 10% declared having seen a health care provider for this reason, even if more than 70% reported contact with a physician within the last 12 months. CONCLUSION: These results show that most adolescents, especially girls, reported recent use of medical services, but did not discuss their health concerns with the doctor. Training should be improved to give better knowledge and counseling skills to health professionals, in order to allow them to address adolescents' health needs. PMID- 10668083 TI - [Preoperative mortality in transposition of the great vessels]. AB - The aim of the study was to report the incidence and causes of preoperative deaths in isolated transposition of the great vessels and to describe the clinical findings in these neonates. PATIENTS AND METHODS: In five French centers of pediatric cardiology, data of all the neonates with isolated transposition of the great vessels who died before arterial switch operations between January 1986 and June 1996 were obtained from reviewing hospital files, echocardiography records and autopsy reports. RESULTS: Among 199 neonates with transposition of the great vessels, 20 (9.9%) died before surgery. The death was related to intracranial haemorrhage in one premature neonate, severe and early hypoxemia in 13 full-term patients (group A) and later sudden collapse in six patients (group B). In group A, the symptoms occurred within 20 minutes after the birth and included cyanosis (n = 12), acute respiratory distress (n = 8), and shock (n = 4). Despite assisted ventilation (n = 13), bicarbonate infusion (n = 12), prostaglandin E1 (n = 7), inotropic drugs (n = 5) and balloon atrioseptostomy (n = 7), death occurred at the median age of five hours. The patent foramen ovale was absent or tiny in ten patients, normal in one patient and not specified in two patients. The ductus arteriosus was patent in ten patients and not specified in three patients. In group B, the neonates were initially in a good hemodynamic condition. Unexplained death occurred between two and five days after the birth: one infant with a large patent foramen ovale did not receive prostaglandin E1, four patients died a few hours after an angiographic study or a balloon atrioseptostomy was performed in a catheterization laboratory, and one child suffered from a cerebral anoxia due to a tightened cord. CONCLUSION: We conclude that the high preoperative mortality rate in isolated transposition of the great vessels is mainly due to absent or small atrial shunt. These findings suggest that only prenatal diagnosis of transposition of the great vessels with immediate balloon atrioseptostomy could avoid a fatal outcome. PMID- 10668084 TI - [Neonatal hemochromatosis]. AB - Neonatal hemochromatosis is characterized by abnormal hepatic and extrahepatic iron overload, which spares the reticuloendothelial system. In neonates, hemochromatosis results in an acute and frequently lethal liver failure. CASE REPORTS: We report five cases of neonatal hemochromatosis which demonstrate various aspects of this disorder and underline the complexity of both the diagnosis and treatment. Case 3 had an extremely low arterial pressure, a presentation not yet described. CONCLUSION: Neonatal hemochromatosis should be suspected in the presence of cholestasis with liver failure of perinatal onset and with high blood level ferritin. Abdominal nuclear magnetic resonance and/or liver biopsy can confirm neonatal hemochromatosis. For one of our patients, a medical treatment allowed us to perform a liver transplantation. PMID- 10668085 TI - [Neonatal hyperparathyroidism secondary to unknown maternal hypoparathyroidism]. AB - Vitamin D or its analogue (calcitriol) is an effective treatment for hypoparathyroidism during pregnancy. Normal babies are delivered and very few cases of associated neonatal hyperparathyroidism are reported. CASE REPORT: We report the case of a baby born to a mother suffering unknown hypoparathyroidism. He was delivered prematurely by cesarean section for birth asphyxia. His mother had had recurrent bouts of impetigo herpetiformis for several months. Severe demineralization associated with decreased plasma calcium level were observed at birth. The PTH level was very high (955 ng/mL) in the baby and very low in the mother, leading to the diagnosis. The baby was given intravenous calcium gluconate, oral 1 alpha-hydroxyvitamin D and mechanical ventilation. Normal plasma calcium level was reached on day 2. PTH level improved by one month of age. Because of impaired respiratory mechanics and bronchopulmonary dysplasia, the baby was kept on ventilation for 58 days. By 20 months of age, he was in good health despite a small height for his age. CONCLUSION: In such cases 1 alpha hydroxyvitamin D seems to be very effective but plasma and urinary calcium levels need careful monitoring. PMID- 10668086 TI - [Preoperative evaluation of choledochal cyst with MR cholangiopancreatography]. AB - The choledochal cyst is a rare congenital disorder usually diagnosed in childhood. It requires a complete surgical resection to prevent complications, particularly the risk of malignant changes. At present, the preoperative examination requires a direct opacification of the biliary tree, but this is an invasive technique with a high risk of infection, especially in pediatric patients. CASE REPORT: A choledochal cyst was diagnosed in a five-year-old girl with recurrent abdominal pain. Diagnosis was made by ultrasound and preoperative evaluation by magnetic resonance-cholangiopancreatography using single-shot fast spin echo sequences. A complete correlation was observed between surgical, preoperative cholangiography and MRCP data. CONCLUSION: Recent improvement in MRCP techniques provide a complete anatomic analysis of choledochal cysts, enabling one to diagnose an anomalous junction of the pancreaticobiliary duct, even the presence of stones within the biliary tree. This short and noninvasive examination should in the future replace direct opacification of the biliary tree for the preoperative assessment of choledochal cysts. PMID- 10668087 TI - [(P)FAPA syndrome: value of cimetidine]. AB - The (P)FAPA syndrome (periodic fever, adenitidis, pharyngitis, aphthous stomatitis) was described in 1987. The etiology of this periodic syndrome remains unknown. We report three new cases. CASE REPORTS: Three girls, aged from 23 months to eight years, developed (P)FAPA. The other causes of periodic fevers were eliminated and the various treatments (antibiotics, antipyretics, nonsteroidal anti-inflammatory agents) proved ineffective. The repetition of the periodic bouts resulted in depressive disorders, absenteeism from school and a drop in weight in the youngest patient. Two of them suffered a sinusal involvement (chronic sinusitis, polyp) and had an increase in the level of immunoglobulin A. In all three cases, cimetidine at a dose of 20 mg/kg/d was well tolerated and resulted in a disappearance of the periodic fevers. CONCLUSION: Cimetidine, as an immunomodulating agent, appears to be beneficial in the in depth treatment of (P)FAPA syndrome. PMID- 10668088 TI - [Palliative care in pediatric oncology]. AB - Improving the management of dying children has always been a common desire among staff who take care of children with incurable life-threatening diseases. Pediatric oncologists are beginning to structure their practice based upon the approach to palliative care given to adults. In the first part of this report, the authors focus on technical care: comfort control and symptoms. The second part is devoted to pain management, a major aspect of pediatric palliative care. In the third part, psychosocial issues are developed, taking into account the point of view of children, siblings, parents and staff. PMID- 10668089 TI - [Iodine nutrition in the infant. Committee on Nutrition of the French Society of Pediatrics]. AB - Iodine is a trace element essential for the synthesis of thyroid hormones. It is present in the human body in minute amounts (15-20 mg in adults). The thyroid is very sensitive to iodine deficiency in newborns and infants because of its very low iodine content. Daily iodine requirements in humans vary from 40 micrograms in neonates to 150 micrograms in adults. Iodine deficiency represents the first cause of avoidable mental deficiency in developed countries; it has not yet disappeared in Europe, especially in the East, where it is responsible for a high prevalence of goiter. Iodine deficiency during pregnancy increases the risk of neonatal transient hypothyroidism, with a high recall rate in programs of systematic screening for congenital hypothyroidism. Data available in France suggest that screening for iodine deficiency should be performed during pregnancy, and that the minimal iodine concentration in formula milk should be increased to 10 micrograms/100 kcal for term infants and 20 micrograms/100 kcal for premature infants. Iodine deficiency is ideally prevented by the use of iodized salt. Because of the risk of iodine overexposure and secondary transient hypothyroidism, the use of iodinated antiseptics must be avoided in premature babies and neonates as well as in pregnant and lactating women. The fight against iodine deficiency, associated with oral stable preventive iodine administration, decreases sharply the risk of thyroid cancer in case of nuclear exposure, by diminishing thyroid uptake of iodine radioactive isotopes. PMID- 10668090 TI - [Radiology case of the month. Extrapontine myelinolysis]. PMID- 10668091 TI - [What is the role of tonsillectomy in children?]. AB - Tonsillectomy is mandatory in the case of chronic airway obstruction associated with tonsillar hypertrophy and in the case of a unilateral enlarged tonsil which may be malignant. Tonsillectomy may be indicated in recurrent tonsillitis. More rarely, tonsillectomy is performed in children for tonsil bleeding or chronic tonsillitis. Allergy and bleeding disorders are not a contraindication for tonsillectomy. PMID- 10668092 TI - [Accidental HIV exposure of children through injury with discarded syringes]. AB - Potential HIV exposure through injury with discarded syringes is a frequent event among children. Although no case of HIV infection has been reported so far in this context, an antiretroviral prophylaxis is generally proposed. Such treatment is extrapolated from consensus for management of exposed health care workers. Extremely low-risk and difficult administration of available drugs allows a case by-case analysis and generally a less aggressive treatment. PMID- 10668093 TI - [Photosensitive epilepsy and television epilepsy]. AB - Photosensitivity is defined by the appearance of occipital or more diffuse electroencephalographic spikes and waves induced by intermittent light stimulation (ILS), particular patterns, TV-watching, and video games. Photosensitivity is a genetic characteristic. Only the diffuse spikes and waves induced by ILS are correlated with epilepsy. Pure photogenic epilepsy is characterized by seizures which are only visually induced, usually by watching TV. Video games sometimes add a trigger effect due to slowly moving patterns or intense brightness. Several epileptic syndromes are associated with a photosensitivity with or without visually-induced seizures, mainly generalized idiopathic epilepsy. PMID- 10668094 TI - [Spontaneous neonatal gastric perforation]. PMID- 10668095 TI - Transient reactivation of CSF in parthenogenetic one-cell mouse embryos. AB - During meiosis, the cytostatic factor (CSF) activity stabilizes the activity of the M-phase promoting factor (MPF) in metaphase II arrested vertebrate oocytes. Upon oocyte activation, the inactivation of both MPF and CSF enables the entry into the first embryonic mitotic cell cycle. Using a biological assay based on cell-fusion (hybrid between a parthenogenetically activated egg entering the first mitotic division and an activated oocyte), we observed that in activated mouse oocytes a first drop in CSF activity is detectable as early as 20 min post activation. This suggests that CSF is inactivated upon MPF inactivation. However, CSF activity increases again to reach a maximum 60 min post-activation and gradually disappears during the following 40 min. Thus, in activated mouse oocytes (undergoing the transition to interphase) CSF activity fluctuates before definitive inactivation. We found that hybrids arrested in M-phase, thus containing CSF activity after oocyte activation, have activated forms of MAP kinases while hybrids in interphase have inactive forms of these enzymes. We postulate that CSF inactivation in mouse oocytes proceeds in two steps. The initial inactivation of CSF, required for MPF inactivation, is transient and does not require MAP kinase inactivation. The final inactivation of CSF, required for normal embryonic cell cycle progression, is dependent upon the inactivation of MAP kinases. PMID- 10668096 TI - The Saccharomyces cerevisiae Cdc14 phosphatase is implicated in the structural organization of the nucleolus. AB - Cdc14, a dual-specificity protein phosphatase, has been previously implicated in triggering exit from mitosis in the yeast Saccharomyces cerevisiae. Using immunofluorescence microscopy and immunogold labeling, we demonstrate that a functional HA-tagged version of the phosphatase Cdc14 localizes to the nucleolus. Moreover, Cdc14-HA co-localized with the nucleolar NOP2 and GAR1 proteins. By immunofluorescence, Cdc14-HA was found in the nucleolus during most of the mitotic cell cycle, except during anaphase-telophase when it redistributed along the mitotic spindle. While this work was in progress, the same pattern of Cdc14 localization was described by others (Visintin et al, Nature 398 (1999) 818). Constitutive overexpression of CDC14 was toxic and led to cell cycle arrest of cells, mainly in G1. This correlated with the appearance of abnormal nuclear structures. A genetic search for suppressors of the lethality associated with CDC14 overexpression identified YJL076W. Because overproduction of Yj1076w buffered the toxic effect of Cdc14 overproduction, this suggested that it might be a substrate of Cdc14. This has indeed been found to be the case by others who recently described Yj1076w/Netl as a nucleolar protein that physically associates with Cdc14 (Shou et al, Cell 97 (1999) 233). The present data confirm several recently uncovered aspects of the regulation of Cdc14 localization and activity and suggest that the level of expression of CDC14 influences the structural organization of the nucleolus. PMID- 10668097 TI - p53-independent regulation of cyclin B1 in normal human fibroblasts during UV induced G2-arrest. AB - Recently we demonstrated, using normal human fibroblasts (NHFs), that UVc radiation induces a G2/M arrest which was even more pronounced when p53 expression was inhibited. So, the aim of this study was to evaluate in NHFs the relationship between UV-induced G2/M arrest and cyclin B1 regulation and to investigate if p53 could contribute to the cyclin B1 regulation in these conditions. Following exposure of asynchronous NHFs to UV light, we showed that the induced G2/M arrest was accompanied by a dose-dependent down-regulation of cyclin B1 mRNA as evaluated by RT-PCR. Concomitantly, using flow cytometric analysis, we observed a strong accumulation of cyclin B1 protein which was correlated to the apparition of the G2/M arrest. In order to study the contribution of p53 to the cyclin B1 accumulation in response to UV exposure, we inhibited p53 induction using p53 antisense oligonucleotides. We found that the inhibition of p53 protein induction after UV exposure had no effect on the level of cyclin B1 mRNA. Moreover, although inhibition of p53 protein induction increased the number of the cells in the G2-M phase, the mean content of cyclin B1 protein was not augmented in these cells. These results indicate clearly that the induction of p53 protein following UV exposure does not regulate the level of cyclin B1 mRNA or protein in normal cells. PMID- 10668098 TI - Dynamics of glial fibrillary acidic protein distribution in cultured glomerular podocytes and mesangial cells of the rat kidney. AB - Glial fibrillary acidic protein (GFAP) has recently been shown to be expressed in the glomerular podocytes and mesangial cells (MC) of kidney (Buniatian et al (1998) Biol Cell 90, 53-61). The different localization of GFAP in podocytes and MC has raised the question whether this might reflect specific cellular functions. To address this question, in the present study podocytes and MC in early (2, 3 day-old), prolonged (5, 7 day-old) and late (14, 21 day-old) primary cultures from out-growths of glomerular explants were used. Double-immunolabeling studies demonstrated that podocytes transiently acquire myofibroblastic features, characterized by the expression of smooth muscle alpha-actin (SMAA) and increased perinuclear reaction of GFAP in prolonged cultures. The morphological differentiation of cobblestone-like podocytes into process-bearing cells was followed by loss of the myofibroblastic marker, SMAA, de novo expression of desmin, and distribution of GFAP, vimentin and desmin into the processes. In late culture, GFAP and SMAA were nearly absent from the podocytes which maintained the cobblestone-like morphology. By contrast, the myofibroblastic features gained by MC during prolonged culturing increased with time. A myofibroblast-like cytoskeleton of podocytes and MC similar to that of healthy astrocytes suggest an increased spectrum of functional activities of these cells during the acquisition of myofibroblastic features. In addition, the present study provides a new combination of biochemical and biological features by which podocytes and MC can be distinguished in culture. PMID- 10668099 TI - Composition and organization of tubulin isoforms reveals a variety of axonemal models. AB - In the flagellum of mammalian spermatozoa, glutamylated and glycylated tubulin isoforms are detected according to longitudinal gradients and preferentially in axonemal doublets 1-5-6 and 3-8, respectively. This suggested a role for these tubulin isoforms in the regulation of flagellar beating. In the present work, using antibodies directed against various tubulin isoforms and quantitative immunogold analysis, we aimed at investigating whether the particular accessibility of tubulin isoforms in the mammalian sperm flagellum is restricted to this model of axoneme surrounded with periaxonemal structures or is also displayed in naked axonemes. In rodent lung ciliated cells, all studied tubulin isoforms are uniformly distributed in all axonemal microtubules with a unique deficiency of glutamylated tubulin in the transitional region. A similar distribution of tubulin isoforms is observed in cilia of Paramecium, except for a decreasing gradient of glutamylated tubulin labeling in the proximal part of axonemal microtubules. In the sea urchin sperm flagellum, predominant labeling of tyrosinated and detyrosinated tubulin in 1-5-6 and 3-8 doublets, respectively, were observed together with decreasing proximo-distal gradients of glutamylated and polyglycylated tubulin labeling and an increasing gradient of monoglycylated tubulin labeling. In flagella of Chlamydomonas, the glutamylated and glycylated tubulin isoforms are detected at low levels. Our results show a specific composition and organization of tubulin isoforms in different models of cilia and flagella, suggesting various models of functional organization and beating regulation of the axoneme. PMID- 10668100 TI - Rat arylalkylamine N-acetyltransferase gene: upstream and intronic components of a bipartite promoter. AB - The daily rhythm in the activity of arylalkylamine N-acetyltransferase (AA-NAT) controls the rhythm in melatonin synthesis in the pineal gland. In the rat, transcriptional regulatory mechanisms play a major role in determining the observed pattern of AA-NAT gene expression. Remarkably, high levels of AA-NAT transcripts can only be detected in the night pineal; significant levels can also be found in the retina. To characterize the regulatory events that impinge upon the activity of the AA-NAT gene we embarked on the systematic analysis of the AA NAT promoter. To this end we transfected several AA-NAT promoter derivative constructs to monitor reporter gene activity in both pineal and non-pineal primary cell cultures. Our studies revealed a cooperative arrangement between upstream promoter and downstream intronic regions which appear to contain most of the key elements necessary to ensure the proper spatio-temporal pattern of AA-NAT gene expression. PMID- 10668101 TI - Methodological aspects of auditory evoked potentials. PMID- 10668102 TI - Transformed migraine: a proposal for the modification of its diagnostic criteria based on recent epidemiological data. PMID- 10668103 TI - Possible antimigraine mechanisms of action of the 5HT1F receptor agonist LY334370. AB - This study investigated whether the selective 5HT1F receptor agonist LY334370 has other possible antimigraine mechanisms in addition to the proposed inhibition of dural plasma extravasation. LY334370 (up to 10(-5) M) had no vasoconstrictor effects on human cerebral arteries in vitro. It had no effect (up to 10 mg kg-1, i.v.) on neurogenic vasodilation of dural blood vessels produced by electrical stimulation of the dura mater in anesthetized rats. Nor had it any effect (at 3 mg kg-1, i.v.) on the hyperalgesia produced by injection of carrageenan into the paw of conscious rats or on nociceptive reflex responses in the spinalized, decerebrate rabbit (up to 3 mg kg-1, i.v.), indicating that it has no general analgesic properties. However, it significantly inhibited activation of second order neurons in the trigeminal nucleus caudalis produced by electrical stimulation of the dura mater in anesthetised rats at 3 mg kg-1, i.v. These results provide evidence to suggest that LY334370 has a central mechanism of action in blocking the transmission of nociceptive impulses within the trigeminal nucleus caudalis and that this may represent a mechanism through which it has its antimigraine effect. PMID- 10668104 TI - Serotonin 5HT1B/1D receptor agonists abolish NMDA receptor-evoked enhancement of nitric oxide synthase activity and cGMP concentration in brain cortex slices. AB - Our previous studies indicating that the function of excitatory amino acids, NMDA type receptor, is modulated by serotonin focused on the interaction between serotonin 5HT1B/1D and glutamate, NMDA receptor in brain cortex. The effect of agonists of 5HT1B/1D receptor, sumatriptan, and zolmitriptan on NMDA receptor evoked activation of nitric oxide (NO) and cGMP synthesis in adult rat brain cortex slices was investigated. Two kinds of experiment were carried out using adult rats. In one of them, sumatriptan or zolmitriptan was administered in vivo subcutaneously (s.c.) in a dose of 0.1 mg per kg body weight. Brain slices were then prepared and used in the experiments or, in the other exclusively in vitro studies, both agonists at 10 microM concentration were added directly to the incubation medium containing adult rat brain cortex slices. The data obtained from these studies indicated that stimulation of NMDA receptor in brain cortex slices leads to a large increase in calcium, calmodulin-dependent NO synthase (NOS) activity and to significant enhancement of the cGMP level. This NMDA receptor-dependent NO and cGMP release was completely blocked by competitive and noncompetitive NMDA receptor antagonists APV (10 microM) or MK-801 (10 microM.), respectively. The specific inhibitor of Ca(2+)-dependent isoforms of NOS (N-nitro 1-arginine NNLA and 7-nitroindozole (7-N1)) eliminated the NMDA receptor-mediated enhancement of NO and cGMP release. Moreover, the serotonin 5HT1B/1D receptor agonists sumatriptan and zolmitriptan administrated in vivo (s.c.) or in vitro abolished NMDA receptor-evoked NO signalling in brain cortex. The potency of both agonists investigated directly in vitro was similar to their effect after in vivo administration. These results suggest that both serotonin 5HT1B/1D receptor agonists may play an important role in modulating the NO and cGMP-dependent signal transduction pathway in the brain. This effect of sumatriptan and zolmitriptan on NO signaling in the brain system should be taken into consideration when investigating their mechanism of action in the migraine attack. PMID- 10668105 TI - Genetic and environmental influences on recurrent headaches in eight to nine-year old twins. AB - The major objective of the present study was to estimate genetic and environmental influences on recurrent headaches in prepubescent twins. A nationwide cohort of 8 to 9-year-old Swedish twins (n = 1,480) was screened for nonsymptomatic and recurrent headaches through a questionnaire mailed to their parents (75% response frequency). Among positives, 79% of headaches were classified as migraine or tension-type headache in close accordance with the International Headache Society criteria. The prevalences of migraine and nonmigrainous headaches were 2.4% and 11.3%, respectively, and without significant differences between the sexes or zygosity types. Inheritance on liability to recurrent headaches was estimated to 70% (a2 = 0.70, 95% CI = 0.54 0.82) for boys and girls but the genetic effect was found to be uncorrelated between the sexes. We conclude that genetic and individual specific effects are important for recurrent headaches of migrainous and nonmigrainous types in prepubescent children, and that different genetic etiologies might exist for boys and girls. PMID- 10668106 TI - Repeatability of the intensity dependence of cortical auditory evoked potentials in the assessment of cortical information processing. AB - Data on repeatability and optimal settings are needed when studying the influence of drugs on the intensity dependence of auditory evoked cortical potentials (IDAP). IDAP was recorded at intervals of 1, 2, and 24 h at two centers in 22 healthy volunteers. Settings were modified to compare fixed versus randomly varied stimulus repetition rate, as well as 30 Hz and 100 Hz low pass filters. Repeatability was assessed for different intervals and different settings. Group means did not differ between centers, the 2-h or 24-h retest, or when using different settings. We observed an order effect for the 1 h retest. Fixed repetition rate and the 30 Hz filter improved repeatability with still high intraindividual variability. IDAP group means can be compared between centers for retest intervals of 2 h and 24 h and different settings. Variability is too large to compare individuals. PMID- 10668107 TI - Auditory evoked potentials in the assessment of central nervous system effects of antimigraine drugs. AB - Because the "intensity dependence" of cortical auditory evoked potentials (IDAP) is under serotonergic control, it can be used to assess central antimigraine effects of 5HT1B/1D agonists. We measured IDAP before and 2 h after naratriptan (5 mg, n = 19) and zolmitriptan (5 mg, n = 19) in healthy volunteers. IDAP was expressed as the amplitude-stimulus intensity function ("ASF slope"). Naratriptan tended to increase ASF slope (mean difference 0.23 +/- 0.62 microV/10 dB, p = 0.06) while zolmitriptan (0.08 +/- 0.95 microV/10 dB, p = 0.35) did not. We assessed the suitability of IDAP for measuring central antimigraine drug effects using repeatability data (see companion paper). We calculated the trade-off between the size of the expected drug effects (ASF slope difference) and the necessary sample size. Because of poor repeatability 36 to 80 subjects are required to detect ASF slope changes in the 0.25-0.5 microV/10 dB range. These data can be used to design trials using IDAP. PMID- 10668108 TI - Out-of-body experiences and related phenomena in migraine art. AB - In a collection of 562 migraine art pictures, seven pieces illustrate various elements of out-of-body experiences (OBEs) and related phenomena, including the somesthetic sensations of a duplicate or parasomatic body and the visual experiences of perceiving the own body, i.e. autoscopy, and its environment from a vantage point out of the body. Phenomenological features of the OBEs depicted are compared with 17 similar case reports reviewed from the literature. It is concluded that OBEs can occur as migraine aura symptom, which supports the notion that OBEs represent a preformed functional response of the brain. This neuropsychological theory supplements existing psychological theories of OBEs, which consider the said phenomena as representing hallucinatory experiences based on imagination and memory. PMID- 10668109 TI - Cervicogenic headache: long-term postoperative follow-up. AB - The patient, a 50-year old female had been suffering from right-sided head- and neck pain since she was 31 years of age. It started in connection with an indirect neck trauma. Analgesics were of little or no avail and operative procedures, including liberation of the greater occipital nerve (GON) (n = 2) and decompression of the C2 ganglion/root, had only a transitory effect. At 42, a magnetic resonance scan of the cervical spine demonstrated a degenerated disk C5 C6, with encroachment on the foramina and the cord. At 42 years of age, a stabilization operation at C5-C6 (Robinson-Smith) alleviated her discomfort--only some motor complaints in the ipsilateral upper extremity remaining and only in the first 12-18 months. PMID- 10668111 TI - The tension-type headache alternative. Peripheral pathophysiological mechanisms. PMID- 10668110 TI - Muscle activity as a releasing factor for pain in the shoulder and neck. AB - In this review, the evidence for trapezius muscle activity as a releasing factor for shoulder and neck pain is considered, mainly on the basis of studies in our laboratory. Two lines of evidence are produced, (i) vocational studies in an occupational setting, where muscle activity pattern is recorded by surface EMG and a clinical examination of the shoulder region of the subjects performed; and (ii) laboratory studies where muscle activity patterns and pain development are recorded in an experimental situation with mental stress and minimal physical activity. The vocational studies demonstrate pain development in the shoulder and neck despite very low muscle activity recorded, making it very difficult to assume muscular involvement for all cases with such complaints. However, the hypothesis of pain development through overexertion of a subpopulation of low threshold motor units also makes it difficult to draw a firm negative conclusion. The laboratory experiments, on the other hand, show that trapezius activity patterns in response to stress have many features that would be expected if muscle activation induces pain symptoms. It is further noted that the trapezius is the only muscle with activity patterns that show these features. Possibly, we observe the effects of parallel physiological phenomena, e.g., a systemic autonomic activation that induces pain symptoms and also facilitates the motor response of some muscles. Evidence of autonomic activation of trapezius is presented by the observation of low-level, rhythmic EMG activity during sleep. However, this is not firm evidence for the above hypothesis, which at present best serves as a basis for further experimentation. PMID- 10668112 TI - Cervicogenic headache: a comparison with migraine and tension-type headache. AB - Cervicogenic headache (CEH) is a neck-generated headache syndrome. Attacks may be similar to migraine (M) or tension-type headache (TTH). In order to test the accuracy of the IHS diagnostic criteria for M and episodic TTH and of the criteria for CEH of Sjaastad et al., 33 CEH, 65 M, and 29 TTH were evaluated according to the CEH criteria, and CEH patients were tested for M and TTH according to the IHS criteria. Only 30% of the CEH patients met the criteria for M, 3% met the criteria for TTH, and 66% were neither M nor TTH. The mean number of criteria met, sex, age, and age of onset were also analysed, and the results indicate an inequality among these three headache types. The most important differentiating aspects were the site and radiation of the pain, the temporal pattern, and the induction of attacks from neck posture, movements, and/or digital pressure. CEH clearly differs from M and TTH. Existing criteria adequately distinguish the three headaches. PMID- 10668113 TI - Migraine and hypertension. AB - Association between mild to moderate hypertension and headache is probably coincidental. Severe sustained hypertension, malignant hypertension and paroxysmal hypertension (sudden rise) are associated with severe headache. Transient hypertension can occur during an attack of migraine or cluster headache. Hypertension may increase the frequency and severity of migraine in migraineurs and may transform an episodic migraine into chronic daily headache. Concomitant treatment of hypertension is important in these patients. PMID- 10668114 TI - Epidemiology and socio-economic impact of headache. AB - Headache disorders constitute a public-health problem of enormous proportions, with an impact on both the individual sufferer and society. Epidemiological knowledge is required to quantitate the significance of these disorders. The effects on individuals can be assessed by examining prevalence, distribution, attack frequency and duration, and headache-related disability. The socio economic burden includes both direct costs associated with healthcare utilization and costs associated with missed work due to sickness absence or reduced efficiency. The individual and socio-economic burden of headaches is substantial. Headache disorders deserve more attention, especially concerning strategies leading to adequate primary prevention, diagnosis, and treatment. PMID- 10668115 TI - The Vaga Study: an outline of the design. AB - We carried out an epidemiological study of headache in a rural parish in the mountainous region of southern Norway. During a 2-year period from October 1995, 1838 parishioners in the age range 18-65 (or 88.6% of the target group) were examined in a structured interview based on a questionnaire. The questionnaires were validated in two ways: 100 records were re-checked, with a consistency of 98%, and a re-check of 41 parishioners was carried out > 2 months after the 1st examination. The details of the latter control study will be reported later. Only one result of the study is given: namely, idiopathic stabbing headache ("jabs and jolts syndrome") was present in > 30% of parishioners. PMID- 10668116 TI - Periorbital pain: a clinical review. PMID- 10668117 TI - Recurrent Tolosa-Hunt syndrome: a report of ten new cases. AB - Ten patients (6F, 4M) with recurrent Tolosa-Hunt syndrome are reported. Besides ocular motor symptoms, one patient had trigeminal nerve involvement, one had ipsilateral ocular sympathicoplegia with miosis and ptosis, and one tinnitus during an episode of Tolosa-Hunt syndrome, ipsilateral to the pain side. One patient had Bell's palsy, one had a possible Raeder's syndrome, and one had a period of tinnitus between the Tolosa-Hunt syndrome episodes. Three of the 10 patients reported periods of periocular pain without ophthalmoplegia between the Tolosa-Hunt episodes, the pain located ipsilateral to the ophthalmoplegic side in the Tolosa-Hunt episodes. Systemic symptoms associated with Tolosa-Hunt syndrome, e.g., back pain, chronic fatigue, arthralgia, gut problems among others, occurred with the same frequency in these 10 patients as in an earlier report. Seventy per cent of the patients had signs of inflammation in serum during a period of Tolosa Hunt syndrome. Orbital phlebograms showed pathologic signs in four of the five patients investigated during a Tolosa-Hunt period. One phlebogram was normal in a sixth patient when performed during a period of unilateral periocular pain without ophthalmolegia. Magnetic resonance imaging of the head (with contrast) was only performed in three patients during the Tolosa-Hunt period: one showed signs of inflammation in the middle fossa and two were normal. In one of the patients with normal magnetic resonance imaging, the orbital phlebogram was pathologic. Steroid treatment promptly relieved the pain in all patients. PMID- 10668118 TI - Tolosa-Hunt syndrome: focus on MRI diagnosis. AB - Experience with modern neuroimaging techniques, computed tomography (CT) and magnetic resonance imaging (MRI) scans, in the diagnosis of Tolosa-Hunt syndrome (THS) is reviewed. Conventional CT scan remains normal in about two-thirds of these patients. In the reported 22 patients meeting the IHS criteria for a THS diagnosis on whom an MRI study was performed, MRI revealed a convex enlargement of the symptomatic cavernous sinus by an abnormal tissue isointense with gray matter on short TR/TE images and isohypointense on long TR/TE images. This abnormal tissue markedly increases in signal intensity after contrast injection. MRI seems also to be the ideal technique to follow progressive resolution of the abnormal tissue after steroids. Therefore, normal MRI would probably exclude THS, whereas in the appropriate clinical setting of steroid-responsive painful ophthalmoplegia, MRI showing the cavernous sinus abnormality described here suggests a diagnosis of THS. From these data, we propose that the fourth IHS criterion for THS diagnosis, "Exclusion of other causative lesions by neuroimaging and (not compulsory) carotid angiogram" should be changed to "Finding by MRI of specific cavernous sinus abnormalities (with the characteristics described herein) which slowly resolve with steroid treatment". PMID- 10668119 TI - Cluster headache: new perspectives. PMID- 10668120 TI - Raeder's syndrome. AB - Raeder's syndrome was first described by the Norwegian ophthalmologist J. G. Raeder in 1918, and the description extended in 1924 by the same author. The seminal report was a description of a young, male patient with unilateral periocular pain combined with ipsilateral miosis and ptosis, and with slight objective signs of trigeminal nerve involvement. Autopsy demonstrated a tumor at the base of the skull in the middle cranial fossa. The term "paratrigeminal" was coined for the picture reported. Later case reports by the same and other authors have included patients with a more benign clinical course, including spontaneous remissions, with unilateral periocular pain and ipsilateral signs of oculosympathetic paresis as the common denominator. This review is a chronological survey of the main contributions that have appeared in the literature and an outline of the various definitions of the syndrome, including a recent classification as well as some pathophysiological and prognostic considerations. PMID- 10668121 TI - SUNCT syndrome versus idiopathic stabbing headache (jabs and jolts syndrome). AB - Both SUNCT syndrome and idiopathic stabbing headache (ISH) (jabs and jolts syndrome) have to be considered when encountering shortlasting headaches. Since there are no specific tests for these headaches, the differential diagnosis depends entirely upon assessment of the clinical features. These headaches are generally easily distinguishable clinically. There seem to be symptomatic forms of SUNCT. PMID- 10668122 TI - Familial occurrence of headache. AB - Associations between the occurrences of headache among parents and their offspring during a 7-year follow-up were studied. Data were collected using a prestructured questionnaire from a representative population-based sample of 1443 families expecting their first child. Seven years later, another questionnaire was sent to 1132 families still included in the study. Questionnaires were returned by 968 families. One or either of the parents had experienced frequent headache in 47% of families (34% of the mothers and 19% of the fathers) before pregnancy. Of the 6-year-old children, 15% had headache disturbing daily activities. Mother's prepregnancy headache was a clear predictor of her child's preschool headache (p = 0.006, OR = 1.7, 95% CI 1.2-2.4). In the clinical interview, the children with headache more often had first-degree and second degree relatives with headache than the control children. PMID- 10668123 TI - Migraine and nonmigrainous headache--how to distinguish them. PMID- 10668124 TI - Tension-type headache in children. PMID- 10668125 TI - Sleep hygiene and migraine in children and adolescents. AB - Although sleep problems are a common complaint in migraine patients, the role of sleep habits and hygiene as triggering factors of head pain attacks has been poorly analyzed. The aim of this study was to evaluate the effect of modifying bad sleep habits across several headache parameters. Based on our previous study, we selected 70/164 migraineurs (42.7%) with poor sleep hygiene and randomly assigned them to two groups: group A migraineurs, who were instructed to follow directions to improve sleep hygiene; and group B migraineurs who were not given instructions on improving sleep hygiene. Mean duration and frequency of migraine attacks were significantly reduced at follow-up in group A, while group B showed only an insignificant initial reduction. No differences were found in the severity of migraine attacks that seemed related to a higher prevalence of nocturnal symptoms such as bedtime struggles, hypnic jerks, nightmares, and restless sleep. Our study is an alternative approach to the treatment of migraine, i.e. treatment through a simple modification of sleep behavior without recurring to pharmacological treatment. PMID- 10668126 TI - Slow cortical potentials in migraine: a comparison of adults and children. AB - Amplitudes of contingent negative variation (CNV) in 40 migraine patients were studied during the pain-free period and compared with healthy controls (n = 24). The early component (initial CNV) is especially more negative in migraine patients. Within the migraine group, 14 children with migraine without aura (mean age 13.6 years) were compared with 11 healthy children (mean age 12.4 years). Migraine children differ from healthy children in the same manner that adult migraine patients differ from healthy adults. The CNV of five healthy children did not differ from the CNV of their siblings suffering from migraine. We suggest that there is a family-related cortical hypersensitivity which does not necessarily lead to the development of migraine. PMID- 10668127 TI - Acupuncture in headache. PMID- 10668128 TI - Insecticidal activity of transgenic tobacco plants expressing both Bt and CpTI genes on cotton bollworm (Helicoverpa armigera). AB - Transgenic tobacco plants expressing both Bacillus thuringiensis (Bt) insecticidal protein and cowpea trypsin inhibitor (CpTI) genes was used to evaluate the insecticidal activity on the cotton bollworm (Helicoverpa armigera) compared with transgenic tobacco with Bt insecticidal protein gene alone. Mortality of first to third instar larvae fed on transgenic two genes tobacco for three days was significantly higher than that fed on transgenic Bt tobacco. First to fourth instar larvae fed on transgenic two genes tobacco continuously could not survive until pupation. Second instar larvae were fed on transgenic tobacco plants for three days and then were transferred to an artificial diet. The efficacy of transgenic two genes tobacco on mortality, larvae weight, percentage of pupation, and time to pupation were significantly higher than that of transgenic tobacco with Bt insecticidal protein gene alone. The results with transgenic tobacco as model plant were valuable for other crops both for the enhancement of insecticidal efficacy and for the delay of insect adaptation to transgenic Bt crops. PMID- 10668129 TI - Cloning of Schistosoma japonicum Chinese strain 22.6kD membrane-associated protein (Sj-22.6) gene and its overproduction on Escherichia coli. AB - A 567bp DNA fragment was amplified from Schistosoma japonicum adult worm mRNA by RT-PCR. Sequence analysis revealed that this fragment contained S. Japonicum Chinese strain membrane-associated protein (Sj-22.6) gene. Then this gene was cloned into the expression vector pGEX-4T, and subsequently expressed in Escherichia coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was about 48 kD. The yield of expression was around 40 mg/L E.coli culture. The immunological test suggested that the recombinant protein had good antigenity which could make a good basis for the research of its immunological function in Schistosomiasis. PMID- 10668130 TI - Purification and properties of genetic expressing product of thermostable protease from Bacillus stearothermophilus HY-69. AB - The thermostable metal protease gene from Bacillus stearothermophilus HY-69 had been cloned and expressed in Bacillus subtilis MI113. The genetic expressing product of the enzyme was purified by CM-cellulose chromatography. The product shows homogeneity on PAGE. Its molecular weight is 27,000 +/- 1000 by SDS-PAGE and Sephadex G100 filtration, respectively. The alpha-helix content of the protease is estimated to be about 66%, the beta-turn about 28%, the random coil about 6%, but not beta-sheet, calculated from the circular dichroism data. The optimal temperature of the enzyme was 70 degrees C. When the enzyme was denatured in 3 mol/L of Gdn--HCl in phosphate buffer pH6.0 for 20 min, it remained about 40% of original activity. It shows that it is rather resistant to heat and Gdn HCl denaturation. Its conformational variety coursed by Gdn-HCl was investigated by the far UV circular dichroism and fluorescence spectra. The results show that the enzyme has more compact conformation and internal hydrophobility. PMID- 10668131 TI - High level expression of the deleted and mutated urokinase-scFv fusion gene in Escherichia coli. AB - The fusion gene of a specific anti-human fibrinogen D-dimer scFv and low molecular weight single chain urokinase (scu-PA-32K) was restricted, spliced, and digested with exonuclease Bal31 to obtain a series of deletion mutants. Study of their expressions in E.coli revealed that the key sequence which reduced its expression level resided in the fragment from 841 to 851 bp, in which a tandem of AGG codons (encoding arginine, rarely used in E.coli) existed. By means of PCR mediated site-directed mutagenesis, we altered these two AGG codons to CGT codons, which could be more efficiently translated in E.coli, and the expression level turned out to be about 30% of the total bacterial proteins while that of the natural gene was only 2%-3%. PMID- 10668132 TI - Hirudin display on the surface of bacteriophage M13. AB - Hirudin was fused to the N terminus of M13 minor protein gp3 (197-406) through a linker GGGS by inserting both the hirudin gene and the gp8 signal sequence into the modified phagemid vector pCANTAB 5V to construct pCANTAB 5G8-Hir. The expressed fusion protein was directed by gp8 signal peptide into the periplasm and assembled to the phage particle to form the hirudin-phage. The fusion protein and fusion phage were detected with biotin-thrombin by Western blotting analysis. Antithrombin activity analysis confirmed that the hirudin portion in the fusion protein and fusion phage bear similar native conformation. The successful display of hirudin on the surface of M13 phage laid a sound foundation for the further study on directed evolution of antithrombotic proteins with altered properties. PMID- 10668133 TI - Production of chimeric antibody in cells and larvae of silkworm. AB - By using the recombinant baculovirus expression system, the chimeric antibody against human small cell lung carcinoma was expressed in the cells and larvae of silkworm Bomyx mori N (BmN). The chimeric antibodies produced in BmN cells and larvae which were infected with recombinant viruses rNPVL2, rNPVH17 and dual recombinant virus rNPVLH19 were detected by Western blot analysis. The co expression products of heavy- and light-chain genes in silkworm larvae revealed stronger immune binding activity than the single product of heavy or light chain gene expression. PMID- 10668134 TI - Construction of high ergosterol-producing yeast strains and study on the optimal conditions for culture. AB - High ergosterol-producing yeast strains were constructed by primary screening isolation of haploid mutagenesis and protoplasts fusion. The fermentation conditions of fused strain YEF-21 were studied. Under the optimal conditions, i.e., the medium for fermentation consisted of 8% glucose 1% polypeptone 1% yeast extract; initial pH nature; 60 mL broth/250 mL flask; inoculum volume 10%; fermentation time 30 hours at 28 degrees C and 200 r/min; the comprehensive value of biomass and ergosterol content of YEF-21 is 1.54 and 1.55 times that of the parent strains YE227 and YE180, respectively. The fused strain YEF-21 is stable enough genetically according to the analysis of genetic stability, and it is a high ergosterol-producing strain that has prospect for application. PMID- 10668135 TI - Studies on the callus cultures of Ginkgo biloba and its metabolites-ginkgolides. AB - The production of ginkgolides in callus culture of Ginkgo biloba was reported. The affection of some physical factors and chemical substances on the induction and growth of calli was also investigated. A biologically quantitative method (platelet aggregation induced by PAF) and HPLC were successfully used for the determination of Ginkgolides A and B in all kinds of callus cultures. The result showed that the content of Ginkgolides B in the callus cultures varies from 0.005% to 0.01%, which is one of the best results for the callus culture of G.biloba in the world. PMID- 10668136 TI - Somatic embryogenesis of Echinodorus orisis L. and the kinetic changes of the endogenous hormones contents during the embryogenetic process. AB - Somatic embryogenesis was achieved in young Leaf cuttings of Echinodorus orisis L., an aquatic ornamental plant, in a short period (25 days). Among the cytokinins and their combinations tested, 6-BA (1 mg/L) and Zt (1 mg/L) in MS medium induced the highest efficiency (100%) of somatic embryogenesis, with a maximum of 4.87 embryoids per explant. Roots instead of somatic embryos were formed when NAA (0.5 mg/L) was added to MS medium containing Zt (1 mg/L). Matured embryoids were germinated and rooted in MS medium with IAA (1 mg/L) after 5 days cultivation. Seventy-two percent of the rooted plantlets transplanted survived in the aquarium. The endogenous hormone contents in various stages of somatic embryogenetic process were measured by HPLC. The concentrations of all the hormones tested were about 2 times that of the cuttings from the untreated fresh leaves after 10 days incubation. Meanwhile, the concentration of IAA presents two peaks after 10 and 25 days of cultivation, respectively. The cytokinin (Zt and ZR) peak, about 8 times more than CK, appeared in 15 days cultivation when the heart-shaped embryos formed. The fluctuation of the GA3 concentration was very similar to that of cytokinin. The ABA, however, remains stable at quite high concentration after 10 days of cultivation. PMID- 10668137 TI - Preparation of GFLV CP-gene cDNA probe labeled by photosensitive biotin and its application in detection of GFLV. AB - The cDNA of the GFLV CP gene amplified from clone pGAB5 by PCR was labeled by photosensitive biotin. The probe sensitivity is 5 pg, and the detectable amount of unlabeled GFLV cDNA was 10 pg by Dot-blot hybridization when the biotin labeled cDNA was used as a probe. The results of detection for GFLV by probe have positive correlation with those of ELISA. The minimum amount of grapevine tissue for detection by probe was 100 times lower than that by ELISA. The seasons and the position of grapevine leaves do not limit application of this method. PMID- 10668138 TI - A comparison of human spermatozoa immunolabeling features using xenogenic reagents for centrosomal proteins. AB - The centrosome is an organelle essential to proper chromosomal migration and normal cell growth. In the human, the centrosome is comprised of two centrioles and the pericentriolar cytosol; its control of embryo cleavage processes derives from its role as a locus for spindle organisation. At fertilisation, it is the human sperm centrosome that is responsible for ordering these processes, as the oocyte appears not to contain working centrosomal structures. Abnormalities in fertilisation or early embryo cleavage could be related to impaired sperm centrosome structure or function in some cases. While potential future treatments of infertility due to a defective centrosome could involve use of a donor centrosome to restore normal cell development, such an approach would depend on accurate localisation of this organelle for subsequent transplantation. To locate centrosomal components in the heads and tails of human spermatozoa, labeling was performed on intact spermatozoa using antibodies of known specificity to highly conserved centrosomal elements. Following general mapping of immunofluorescent signals, unlabeled sperm were dissected to form head/tail sperm fragments which were then separately tested. Distribution of centrosomal proteins in head and tail fragments was assayed for each separation method. Three reagents were compared: 1) rabbit anti-mitotic spindle protein (anti-MSP) antibody, 2) rabbit polyclonal centriole-specific antibodies, and 3) mouse monoclonal anti-MPM-2 (a centrosome phospoprotein) antibody. Of these, anti-MPM-2 antibody appeared to be the most reliable, labeling centrosomal elements in 63% (n = 1,386) of treated spermatozoa. Sequential utilization of n-butylamine to effect head/tail separation followed by anti-MPM-2 antibody labeling was a satisfactory method of centrosome localisation. Microextraction of centrosomes and pericentriolar matrix identified by this method awaits further testing. PMID- 10668139 TI - Ovarian cyst formation and congenital absence of the inferior vena cava: case report. AB - Congenital absence of the inferior vena cava (IVC) is a rare condition that presents clinically as recurrent venous thromboses and leg ulcers. We report an association with painful ovarian cysts in a 25 year old woman. The possible pathophysiology and unique management issues posed by this case are presented. PMID- 10668140 TI - Trisomy 21 fetus co-existent with a partial molar pregnancy: case report. AB - BACKGROUND: Approximately 1 in 1,000 pregnancies in the United States are complicated by the presence of a hydatidiform mole. A Medline search revealed no reported cases of a trisomic fetus co-existent from 1966-1998. We present the case of a patient, initially found to have hypertension, edema, and proteinuria in the first trimester, and later found to have a partial molar gestation co existent with a trisomy 21 infant. CASE REPORT: A 31-year-old female presented to her family practitioner in the first trimester and was found to have hypertension and proteinuria. A thorough work-up by a nephrologist revealed no cause. The patient was transferred to the Maternal-Fetal Medicine Service at 26 weeks' and 1 day estimated gestational age. An amniocentesis revealed the presence of a fetus with trisomy 21. At 27 weeks' and 3 days estimated gestational age, the patient underwent a cesarean delivery for a non-reassuring fetal heart rate. Pathologic examination of the placenta revealed the presence of a partial hydatidiform molar pregnancy. CONCLUSION: The present account represents the first reported case of a fetus with trisomy 21 co-existent with a partial hydatidiform mole. PMID- 10668141 TI - Potential use of bioelectrical impedance analysis in the assessment of edema in pregnancy. AB - OBJECTIVE: To evaluate the validity of Bioelectrical Impedance Analysis (BIA) in the assessment of edema in pregnancy. METHODS: A prospective study of healthy women identified during the first trimester of pregnancy. From a pool of 200 eligible volunteers, BIA was conducted on 90 women during the rest of pregnancy and postpartum period. RESULTS: The values for bioelectrical impedance in normal pregnant women decreased gradually in the course of pregnancy, whereas a more remarkable decrease in the values was noted in eight patients who developed edema. The bioelectrical impedance (BI) changes correlated closely with body weight changes. There was a strong relation between bioelectrical impedance values and the degree of edema. The precedent decrease of the values before the onset of edema was noted in seven of the eight patients with edema. CONCLUSION: BIA can be a useful and practical method for the early detection and quantitative assessment of edema in pregnant women. PMID- 10668142 TI - Maternal and neonatal outcome in a monochorionic twin pregnancy complicated by single intrauterine demise. AB - Single fetal death in monochorionic pregnancies is believed to be associated with increased risk of perinatal morbidity and mortality for the living twin and risk of coagulopathy affecting the mother. In this report we present a case of single intrauterine death in a monochorionic twin gestation diagnosed in the 28th week of pregnancy. PMID- 10668143 TI - Effects of conjugated estrogens and progestogen in surgically induced endometriosis in oophorectomized rats. AB - This study aimed to verify whether estrogen replacement alone or associated with progesterone promotes the recurrence of experimental endometriosis in oophorectomized rats. The procedure utilized for endometriosis induction was adapted from the one described by Jones (1984). The rats were castrated three weeks after the induction. Hormonal replacement was started 14 days after the castration and was given for 24 days. One group received estrogen alone, another received estrogen associated with medroxiprogesterone acetate, and a last one received placebo. At the end of this study, the animals who received hormonal medication showed recurrence of the disease. This fact was more evident in the group that received estrogen alone. We concluded that estrogen alone leads to recurrence of endometriosis in oophorectomized rats with surgically induced endometriosis. The association of medroxiprogesterone promotes involutional changes in the implants, and should be added upon the existence of a past history of endometriosis. PMID- 10668144 TI - A prospective study of induction of labor with prostaglandin vaginal gel: ambulatory versus in-patient administration. AB - BACKGROUND: Elective induction of labour is a common obstetrical practice. Dinoprostone (prostaglandin E2 in triacetin base gel) has been shown to be an effective and fairly safe agent for this purpose in inpatient settings. Currently published work does not assess the effectiveness and safety of dinoprostone in an ambulatory setting. OBJECTIVE: To assess the difference between inpatient and outpatient use of dinoprostone for elective induction of labour with regard to effectiveness, safety, length of hospital stay, and patient satisfaction. METHODS: A prospective non-randomized study, in which two groups of low risk obstetrical patients who were undergoing elective induction of labour were studied. The outpatient group was drawn from Regina Health District while the inpatient (control group) was drawn from Saskatoon. The maternal and fetal morbidity was compared in both groups as well as the efficacy, length of hospital stay and degree of patient satisfaction. RESULTS: There were statistically significant reductions in the length of hospital stay and greater patient satisfaction in the outpatient group. No difference was found in efficacy and safety of prostaglandin use. CONCLUSIONS: The findings suggest that ambulatory use of prostaglandin gel for induction of labour reduces the length of hospital stay, and leads to greater patient satisfaction. Further randomized studies with a larger number of patients are needed to evaluate the safety of this agent in an ambulatory setting. PMID- 10668145 TI - In vitro decidualization activity assay of human endometrial stromal cells. AB - To address the need for a simple and reliable system for evaluating the effects of bioactive substances on decidualization of endometrial stromal cells, we designed and tested an assay for measuring decidualization activity. With our assay, it is easy to examine the effects of various bioactive substances on endometrial stromal decidualization. For example, as shown in this study, it is now clear that neither RANTES nor gonadotropin releasing hormone agonist has any effect on endometrial stromal decidualization. Our simple assay has also made it possible to evaluate individual stromal decidualization activities and the regulation of decidualization. PMID- 10668146 TI - Transvaginal sonographic evaluation of endometrial polyps: a comparison with two dimensional and three dimensional contrast sonography. AB - Endometrial polyps are a frequent cause of menorrhagia and are often associated with infertility. Routine transvaginal (TV) ultrasound evaluation may detect intrauterine anomalies. Sonographic evaluation of the endometrium after uterine cavity distention, called hystero-sono-salpingography (HSSG), improves the resolutive effectiveness of TV sonography. Recently the development of three dimensional (3D) scanning of the uterine cavity has provided accurate images. The purpose of this study was to compare conventional 2D and 3D scanning of the uterine cavity with or without saline contrast medium in the detection and evaluation of focal endometrial polyps. Twenty-three patients out of 642 women suggestive for intrauterine anomalies at routine TV sonogram were examined by 2D and 3D sonography before and after intrauterine saline contrast medium. Sonographic appearance was verified by hysteroscopic and histologic evaluation. Two-dimensional TV sonography diagnosed 23 polyps versus 16 confirmed at hysteroscopic and histologic examinations, revealing a specificity of 69.5%; 2D TV HSSG diagnosed 17 polyps, with a specificity of 94.1%; 3D TV sonography diagnosed 18 polyps, with a specificity of 88.8%; 3D HSSG diagnosed 16 polyps according to hysteroscopic and histologic findings, with a specificity of 100%. HSSG has been demonstrated to be an effective and suitable method in the detection of intrauterine anomalies, particularly with 3D sonography. PMID- 10668147 TI - Correlation between histological grading and growth-related factors in human endometrial adenocarcinomas. AB - In order to clarify biological significance of histological grading of malignant tumors, several "growth-related factors" were examined and compared in human endometrial adenocarcinomas with different histological grades. Growth fraction (estimated by Ki-67 immunostain) and cytoplasmic bcl-2 and nuclear p53 overexpression were estimated immunohistochemically in 40 cases of carcinomas with different histological grades. When the grades were divided simply into 3 tiers, G1 (well); G2 (moderately); G3 (poorly differentiated), essentially no differences were found between histological grade and growth fraction. In addition, bcl-2 expression, which is known to negatively affect Ki-67 expression, had no correlation with histological grade. Only nuclear p53 overexpression, known to reflect gene alterations, tended to be more common in the higher grade groups. The results indicate that growth fraction does not correlate well with histological grade of human endometrial adenocarcinoma, as far as examined by Ki 67 and bcl-2 immunostains. The p53 gene status may have some significance in histological grade of human endometrial adenocarcinoma, although its role is not always clearly understood. PMID- 10668148 TI - The management of breech presentation in the last three decades. AB - In order to evaluate the changes in management of breech presentation during the last three decades, a retrospective analysis of the "Alexandra" Hospital records was undertaken. The years, 1965, 1975, 1985, 1995 and finally 1997 were used as pilot years for this purpose. The cesarean section, breech presentation and breech cesarean section rates were calculated for each year. Specific breech cesarean section rates in respect to birth weight and parity as well as perinatal mortality rates were also recorded for each pilot year. The overall cesarean section rate rose from 8.0% in 1965 to 25.2% in 1995 and 25.1% in 1997 while the breech cesarean rate climbed from 16.9% in 1965 to 74.1% in 1995 and 72.3% in 1997, irrespective of birth weight. A trend towards vaginal delivery of breeches in multiparous women till 1985 became less apparent in later years. During the same period, a marked decrease of the perinatal mortality rate was observed from 70.1/1000 in 1965 to 36.6/1000 in 1997. In conclusion, a more than four-fold increase of the breech cesarean section rate was apparently rewarded by a two fold decrease in perinatal mortality. PMID- 10668149 TI - Effectiveness of "core biopsy" by the mammotome device for diagnosis of inflammatory carcinoma. AB - OBJECTIVE: To assess the usefulness of "Mammotome" device for the diagnosis of the inflammatory breast carcinoma. MATERIAL AND METHODS: We studied 6 patients, aged 43-79 years, with clinical evidence of inflammatory breast carcinoma. We compared two sampling techniques, a cytologic one by Fine Needle Aspiration (FNA) and a microhistologic one by "Mammotome". RESULTS: Cytologic sampling by FNA permitted certain diagnosis of malignant lesions in 2 out of 6 cases, while the "Mammotome" device confirmed the correct diagnosis in all 6 considered cases. CONCLUSIONS: The "Mammotome" device proved more useful in the diagnosis of inflammatory breast carcinoma than FNA and it can be a valide alternative to surgical biopsy. PMID- 10668150 TI - Red blood cell zinc protoporphyrin measurement for assessment of peripartum iron deficiency. AB - OBJECTIVE: To study the effectiveness of the rapid red blood cell zinc protorphyrin (RBC-ZPP) test for the detection of women with iron-deficiency anemia in the peripartum period. DESIGN: Blood was drawn prospectively from 150 healthy parturient women upon admission to the labor and delivery room and 72 hours after delivery. Concentration of RBC-ZPP was measured and correlated with hemoglobin level (p = 0.001), mean corpuscular volume (p = 0.002), hematocrit (p = 0.0001), platelet count (p = 0.002), and serum iron (p = 0.0001), serum ferritin (p = 0.0001) and serum transferrin (p = 0.0001) concentrations. RESULTS: RBC-ZPP concentration showed a significant increase from pre-delivery to 72 hours post-delivery. This change correlated significantly with the changes in all the other parameters studied. CONCLUSION: The RBC-ZPP test is a reliable, rapid, easy to-perform, and inexpensive method of screening low-risk women, after uneventful vaginal delivery, for iron deficiency. PMID- 10668151 TI - Laparoscopic ultrasonic operative technique in surgery of women with endometrial cancer: 2 case reports. AB - The aim of this study was to introduce a new laparoscopic ultrasonic technique in the laparoscopy-assisted surgical staging of endometrial cancer. The entire laparoscopic phase of the laparoscopic hysterectomy and pelvic lymph node dissection was performed using a 5 mm ultrasonic scalpel and shears. Ultrasonic activated technology was easy to use and allowed the surgeon to perform laparoscopic hysterectomy and lymphadenectomy close to important pelvic structures safer than in other operative techniques. This is only a case report and a larger study to confirm the advantages of the laparoscopic operative technique in surgery of women with uterine malignancy is needed. PMID- 10668152 TI - Preterm delivery: predictive value of cervico-vaginal fetal fibronectin. AB - OBJECTIVE: This study aimed to evaluate the risk of preterm delivery in the asymptomatic obstetric population of L'Aquila by means of fetal fibronectin immunoassay in cervicovaginal secretions. METHODS: In this prospective study, 60 asymptomatic pregnant women at low-risk for preterm delivery were followed-up. Fetal fibronectin cervical swabs from the esocervix and posterior vaginal fornix were obtained every second week from 24 to 36 weeks of gestation. Fetal fibronectin concentrations were measured by an enzyme-linked immunosorbent assay with a cutoff level set at 50 ng/ml. RESULTS: Twelve patients (20%) had at least one positive fetal fibronectin test result. Six women in our study group (10%) were delivered spontaneously < 37 weeks; 4 of these (66%) had at least one positive fetal fibronectin test result (positive predictive value: 33%; sensitivity: 66%) and 3 of these women (75%) had a positive test result between 24 and 26 weeks. The remaining 8 patients with at least one positive fetal fibronectin test were delivered at term or post-term. Forty-eight women always had negative tests and 46 (95.8%) of these were delivered at term (specificity 82%), whereas 2 (4.2%) were delivered prematurely. The negative predictive value of fetal fibronectin as a predictor of term delivery in this low-risk population in 95% with odds ratio = 11.5 (95% confidence interval 1.44 to 110.4), relative risk = 8 (95% confidence interval 1.38 to 59.2) and Fisher Exact Test p < 0.024. CONCLUSION: In a population of asymptomatic patients at low risk for prematurity, the occurrence of a positive cervical or vaginal fetal fibronectin test result defines a subgroup at increased risk for preterm delivery, mostly at low gestational age. PMID- 10668153 TI - Our experience in laparoscopic diagnosis and management in women with chronic pelvic pain. AB - Chronic pelvic pain (CPP) still remains a serious problem in everyday gynecological practice. The aim of this study was to prospectively estimate the occurrence of pelvic varicosities in women with CPP and also to report our experience in the establishment of diagnosis and management of patients with CPP. We examined 264 premenopausal women aged 18 to 42 years referred to us for chronic constant pelvic pain of at least 6 months duration and with incomplete relief by previous treatments. The women were divided into 4 groups in proportion to their parity. The results of our study demonstrate that pelvic congestion is a common finding in women with chronic pelvic pain especially in multigravidas. Based on our findings we support laparoscopic resetting of the uterus as an option for treatment in patients desiring maintenance of future fertility. PMID- 10668154 TI - A note on the functions of bc1-2 in human solid tumors in situ. AB - Dual functions of bc1-2 product, (1) inhibition of apoptosis and (2) inhibition of cell growth, were analyzed in human endometrial adenocarcinoma tissue using serial tissue sections and immunohistochemical and histochemical procedures. As conclusion, bc1-2 product was speculated to have at least two functions; (1) suppression of apoptosis of cells in G0 phase, and (2) blocking the entrance of G0 cells into cell division cycle. PMID- 10668155 TI - Immunohistochemical evaluation of apoptosis in placentae from normal and intrauterine growth-restricted pregnancies. AB - OBJECTIVE: To investigate the extent of apoptosis within the human placenta in tissues from normotensive term pregnancies and those complicated by intrauterine growth-restriction (IUGR). METHODS: A total of 18 placentas, 10 obtained from uncomplicated term gestations and 8 from intrauterine growth restricted fetuses were included in this study. Apoptosis was identified using a terminal deoxynucleotidyl transferase-mediated deoxyuridine triophosphate nick end labeling technique (TUNEL, Boehringer, Mannheim, Germany) in paraffin-embedded sections. RESULTS: Apoptosis was predominantly detected in the villous trophoblast and stromal tissue. There were no differences in the incidence of apoptosis in different parts of the same placenta. The apoptotic index in placental tissue from uncomplicated pregnancies was 0.93 +/- 0.12. Significantly more apoptotic nuclei were detected in the placental tissue from IUGR gestation (4.2 +/- 2.96, p < 0.01). CONCLUSION: These results might point toward a possible role of apoptosis in the pathophysiology of intrauterine growth-restriction. PMID- 10668156 TI - Relationship between unexplained infertility and human leukocyte antigens and expression of circulating autogeneic and allogeneic antisperm antibodies. AB - PURPOSE: The association between unexplained infertility, human leukocyte antigens (HLA) and expression of circulating antisperm antibodies was studied in 52 couples with unexplained infertility and 15 infertile and 9 fertile couples. METHODOLOGY: Evaluation parameters included Terasaki's HLA microlymphocytotoxicity test, circulating antisperm antibodies using immunofluorescence technique. RESULTS: Fifty-two couples (8.7%) out of 600 consecutive clinic attendants had unexplained infertility. Unexplained infertility was associated with circulating antisperm antibodies (22 versus 13 and 0% for men and 18.5 versus 13 and 0% for women). HLA class 1 B6 and B52 and Cw7 and HLA class 2 DR4 and DR6 and sharing of HLA B6, DR4 and DR6 were found more in couples with unexplained infertility. Combined expression of antisperm antibodies by couples, demonstrated more homozygosity for HLA B6 and DR4. CONCLUSION: Homozygosity for these antigens, B6, DR4 and DR6, may enhance the expression of antisperm antibodies, and cause infertility. PMID- 10668157 TI - Intramuscular versus vaginal administration of progesterone for luteal phase support after in vitro fertilization and embryo transfer. A comparative randomized study. AB - A total of 156 patients were randomly treated with exogenous natural progesterone (intramuscularly, 50 mg/day) and vaginal gel (90 mg/day) P or nothing (Controls) from the day before embryo transfer (ET) for two weeks. In case of positive beta HCG, the treatment was continued for 12 weeks. Plasma P and 17 beta-Estradiol concentrations were estimated and compared with the control not supplemented group. Both treatments were able to increase significantly the luteal plasmatic values of P versus controls. The ongoing pregnancy and the living birth rates per transfer were significantly higher in the patients supplemented with intramuscular P than in those treated with vaginal gel P. The intramuscular natural P appears the most suitable route of administration for luteal phase support in IVF-ET procedures. PMID- 10668158 TI - Disposition of pesticides and toxicants in the human reproductive system in cases of acute poisoning. AB - The aim of this study was to estimate the penetration of some of the pesticides and toxicant substances in the human reproductive system. This knowledge is valuable because of the possible adverse influence of these substances on the human reproduction system and the development of the foetus during pregnancy. The existing data is mainly concerned with the results of experimental studies on animals or epidemiological studies. Here we report data concerning the disposition of several toxic xenobiotics (pesticides and solvents) in the tissues of the human reproductive system as well as in other organs and glands. Data was collected from cases of acute poisonings and derived mostly from autopsy materials. Xenobiotics were found to penetrate sampled tissues such as the testes, ovaries, epididymis, uterus, thyroid gland, as well as other human tissues. Further studies will clarify and confirm peculiarities of the penetration of a wide range of substances in various body tissues and will be the base of the estimation of the role of these toxicants in human reproductive ability and the outcome of pregnancy in humans. PMID- 10668159 TI - D-dimer after delivery in uncomplicated pregnancies. AB - D-dimer is now widely used as a coagulation marker. During pregnancy the D-dimer level increases until term even in uncomplicated pregnancies. The aim of the study was to establish the D-dimer immediately after delivery in uncomplicated pregnancies. A rapid immunoturbidimetric assay for D-dimer determination was employed in 100 consecutive deliveries. D-dimer level increased significantly in all women after delivery (increase from 1 to more than 10 times over the normal range). CONCLUSION: An increase in fibrinolysis is associated with pregnancy and delivery, and D-dimer level must be interpreted only in association with other clinical, laboratory and instrumental methods when pathological conditions (e.g. pulmonary embolism, deep vein thrombosis or disseminated coagulation) are suspected. PMID- 10668160 TI - Usefulness of a 12-month treatment with finasteride in idiophathic and polycystic ovary syndrome-associated hirsutism. AB - OBJECTIVE: Hirsutism is considered as a skin disease due to increased 5 alpha reductase activity in the pilosebaceous unit and finasteride is a drug that inhibits this enzymatic activity. This study showed the effectiveness of a chronic treatment with a selective 5 alpha-reductase inhibitor, finasteride, in idiopathic and PCOS-associated hirsutism. METHODS: Finasteride was administered orally at a daily dose of 5 mg for a period of 12 months to 20 women with IH and 20 women with PCOS. MAIN OUTCOME MEASURES: Each group was submitted to clinical (with Ferriman-Gallwey method) and serum hormonal (FSH, LH, 17 beta-estradiol, total and free T, delta 4-androstenedione, DHEAS; dihydrotestosterone, 3 alpha androstanediol glucuronide) studies at baseline and after 3, 6 and 12 months of treatment. RESULTS: After 3 months of finasteride treatment, a significant decrease in the average hirsutism scores was recorded both in IH (p < 0.0001) and PCOS patients (p < 0.0001). A progressive significant decrease of hirsutism score was observed in IH patients after 6 and 12 months (p < 0.002) and in PCOS patients after 6 but not 12 months. In fact, the maximal therapeutic effect on the hirsutism was obtained after 12 months in the IH and 6 months in PCOS group. PMID- 10668161 TI - Benefits of antibiotic prophylaxis in laparoscopic gynaecological surgery. AB - The prevention of infectious complications by antibiotic prophylaxis has made traditional or laparoscopic surgery much safer but at the same time has contributed to an uncontrolled and often irrational use of every kind of antibiotic. In this study we wanted to show that often mini-invasive surgery like laparoscopy can be practised without the use of antibiotics. Thus, postoperatively several patients undergoing laparoscopic, diagnostic and operative interventions were followed-up. The results showed that subjects without antibiotic therapy did not have any symptomatology that could be ascribed to bacterial infections. In conclusion this study has demonstrated that laparoscopic surgery, especially without any complications, should follow the elementary rules of surgical techniques and surgical asepsis and that antibiotic prophylaxis is not always necessary. PMID- 10668162 TI - Vulvar neoplasia and search for human papillomavirus 16 and 18 genetic information. Short communication. AB - The presence of HPV 16 and 18 is frequent in cases with vulvar carcinomas and intraepithelial neoplasias. PMID- 10668163 TI - Inhibitory effect of exogenous oxytocin on ACTH and cortisol secretion during labour. AB - Complex mechanisms which are still not completely defined, are responsible for the spontaneous onset of labour: an essential role is attributed to endocrine factors. A massive increase, even three times higher than normal physiological values of ACTH and cortisol, has been reported during labour. Similar behaviour has also been recorded for oxytocin at the end of pregnancy as well as during labour. The relationship between oxytocin and the adrenal axis are still debated thus the goal of our study was to attempt to clarify this rapport. Sixty-two women at the end of a term-pregnancy agreed to participate in this study: 46 were innoculated with oxytocin (syntocinon) every 20 minutes for 1 hour; 16 were administered a natural placebo every 20 minutes for 1 hour (control group). ACTH and cortisol values from plasma samples were taken every 20 minutes and analyzed. Our results demonstrated an inhibitory effect of exogenous oxytocin on ACTH and cortisol release. This inhibitory effect, as shown by our results, is time and dose-related. High oxytocin levels, as during exogenous infusion, could induce an effect opposite a normal physiologic one. PMID- 10668164 TI - Penis, bladder and uretral agenesis associated with anorectal malformation in a living male neonate. Case report. AB - Aphallia is a very rare congenital malformation, with an occurrence of 1 in every 30 million births. In the international literature about 75 cases have been indicated as of today. The authors report and discuss the case of one neonate, born from a monoamniotic twin delivery, suffering from agenesis of the penis, anorectal malformation with a fully formed scrotum with 2 normal gonads and absence of bladder and urethra and both kidneys. PMID- 10668165 TI - Outcome of stimulated in vitro fertilisation (SIVF) using clomiphene citrate and human menopausal gonadotropin in different infertility groups. AB - A prospective study was undertaken to evaluate the efficacy of stimulated in vitro fertilization (SIVF) using Clomiphene Citrate and hMG in different infertilities. The analysis was applied to the first 81 cycles over a period of 9 months in the years 1994-1996 in Sheffield Fertility Centre (SFC). The female patients included in this study were under 40 years of age, and their FSH and LH values were < 10IU/L. Mild and moderate male factor infertilities were included. For tubal factor infertility 16 cases were included with an implantation rate of 0%. The unexplained factor infertility included 33 cases with an implantation rate per embryo transfer (ET) of 41%. For male factor infertility there were 18 cases with an implantation rate per ET of 42%. Out of 3 cases in the ovulatory factor group none reached ET with 0% implantation. For multiple factor infertility 11 cases were included with a 0% implantation rate. The overall implantation per embryo transfer was 27%, while the implantation per cycle started was 15%. We conclude that there are certain infertility factors, i.e. unexplained infertility and mild male factor, which can have good results in IVF using CC/hMG only. PMID- 10668166 TI - Hematocolpos by imperforated hymen. Case report. PMID- 10668167 TI - Does maternal drug ingestion cause megacystis microcolon intestinal hypoperistalsis syndrome? II. Bromide trial. AB - PURPOSE: Megacystis Microcolon Intestinal Hypoperistalsis Syndrome (MMIHS) is a congenital disease, and the etiology of the disease is unclear. It is speculated that maternal ingestion of some drugs during pregnancy and degeneration of smooth muscle cells in the fetus may be an etiologic factor. In this study we aimed to investigate the effect of maternal ingestion of bromide on the fetal bladder and colon in pregnant rats. METHOD: We separated animals into a bromide group including 30 rats and a control group with 14 rats. Nothing was given to the control group during pregnancy. Intraperitoneally 8 mg/kg/day bromide was given to the study group from the 6th to 12th day of pregnancy. All of the rats were sacrificed on the 20th day of pregnancy. Histopathological examination of fetal colons and bladders was performed. RESULTS: In the bromide group, an increase in the colon and bladder diameter, an increase in muscle atrophy in the colon and bladder wall, an increase in vacuolar degeneration in the muscles of the bladder and colon wall, and a significant decrease in ganglion cell numbers in the myenteric plexus of the colon and bladder were determined. CONCLUSION: In our rat model, we found histological structural changes in the rats' colon and bladder walls as a result of using bromide on the 6-12th days of pregnancy similar to pathological findings found in some of MMIHS patients' bowels and bladders. PMID- 10668168 TI - Does maternal drug ingestion cause megacystis microcolon intestinal hypoperistalsis syndrome? III. Ethanol trial. AB - PURPOSE: Megacystis Microcolon Intestinal Hypoperistalsis Syndrome (MMIHS) is a congenital disease, and the etiology of the disease is unclear. It is speculated that maternal ingestion of some drugs during pregnancy may be an etiologic factor. In this study we aimed to investigate the effect of maternal ingestion of ethanol on the fetal bladder and colon in pregnant rats. METHODS: We separated animals into an ethanol group including 30 rats and a control group with 14 rats. Nothing was given to the control group during pregnancy. Orogastrically 2 cc/day 30% ethanol was given to the study group from the 6th to 12th day of pregnancy. All of them were sacrificed on the 20th day of pregnancy. Histopathological examination of the fetal colon and bladder was performed. RESULTS: In the ethanol group a significant decrease in the colon and bladder diameter, a significant decrease in the thickness of the colon and bladder wall, an increase in vacuolar degeneration in the muscles of the bladder wall, an increase in connective tissue proliferation among the muscles of the bladder, a significant decrease in the number of ganglion cells in the myenteric plexus of the colon and a significant decrease in the thickness of the bladder tunica muscularis were determined. CONCLUSION: In our rat model we found histological structural changes in the rats' colon and bladder wall similar to a pathological finding found in some of the MMIHS patients' bowel and bladder as a result of using ethanol on the 6th 12th days of pregnancy. PMID- 10668169 TI - Synthesis, binding affinity and selectivity of new beta 1- and beta 2 adrenoceptor blockers. AB - The synthesis of a new series of sesamol derivatives with beta-adrenergic blocking activity is described. The affinity and selectivity of these compounds for beta 1- and beta 2-adrenoceptors were studied in comparison with those of ICI 118551 and propranolol. Some of the synthesized compounds show very good affinity for the beta 2-receptors of rat lung membranes and two of them provide interesting selectivity. PMID- 10668170 TI - Antimycobacterial activity of new ortho-, meta- and para-toluidine derivatives. AB - Novel toluidine derivatives are described. Some of them showed an interesting in vitro activity against Mycobacterium tuberculosis, M. smegmatis, M. marinum, M. gordonae, and M. avium. Some of them were more active than Streptomycin and Isoniazid, which were used as controls, against M. avium and M. gordonae. In particular, we confirm the good activity of biphenyl derivatives. PMID- 10668171 TI - Escin in pharmaceutical oral dosage forms: quantitative densitometric HPTLC determination. AB - A practical and reliable method for the quantitative determination of escin in pharmaceuticals was developed. The TLC-densitometric determination was performed without using spray or dipping reagents in order to provide a more rapid and simple analytical procedure. Chromatographic separations were obtained on analytical HPTLC silica gel plates and the elution was made with 3.5:4.5:2 PropOH ethyl acetate-water (v/v/v). Densitometric analysis was performed directly at 212 nm, corresponding to lambda max of escin obtained by in situ-scanning. A second degree polynomial regression relationship was found between the peak areas and the amounts of the escin standard deposited in the range 1.15-6.90 micrograms. The method is specific, accurate and reliable and was applied successfully to the quantitative determination of escin in commercial samples. PMID- 10668172 TI - Docetaxel in combination with epirubicin in metastatic breast cancer: pharmacokinetic interactions. AB - Epirubicin (75 mg/m2) and docetaxel (75 mg/m2) were administered to 16 patients affected by metastatic breast cancer following two different schedules: (1) docetaxel as infusion administered 1 h after epirubicin administration (schedule A); and (2) docetaxel as infusion immediately (10 min) after the end of epirubicin i.v. bolus administration (schedule B). Experimental non-compartmental analyses such as AUC and Css, were affected very little by the drug combination, irrespective of whether the administration of docetaxel was immediately after the epirubicin bolus (10 min) or delayed (1 h). However, serum levels showed evidence of transient drug interaction: in schedule A, docetaxel infusion was associated with a transient increase of plasma epirubicin in correspondence with Cssmax of docetaxel. Bi-compartmental analysis showed a significant difference in epirubicin clearance between protocols A and B. It is suggested that polysorbate 80, used in minimal amounts to formulate docetaxel, may interfere with epirubicin plasma level. PMID- 10668173 TI - Synthesis of 1,2-benzisothiazole derivatives and investigation of their putative histaminergic activity. AB - Some new 2-(1,2-benzisothiazol-3-yl)ethylamine derivatives were synthesised and their putative histaminergic activity was investigated in in vitro gastrointestinal and cardiac preparations. In the isolated guinea pig duodenum, all the compounds induced a tetrodotoxin- and atropine-sensitive contractile activity, which was minimally affected by mepyramine in the case of the compound 2-(1,2-benzisothiazol-3-yl)ethylamine. In the same tissue, all the compounds were devoid of any H3 receptor agonistic or antagonistic activity, but caused a nicotinic and/or 5-HT3 receptor activation. None of these compounds induced any histamine H2 agonistic or antagonistic activity in the isolated guinea pig gastric mucosa or in the isolated papillary muscle. On this latter substrate, the compound N,N,N-trimethyl-2-(1,2-benzisothiazol-3-yl)ethylammonium iodide induced a positive inotropic activity, apparently due to a release of catecholamines. These results demonstrate the substantial inability of 1,2-benzisothiazole derivatives to interact with histamine receptors in functional tests. These compounds, however, possess gangliomimetic properties, related to the activation of 5HT3 and/or nicotinic receptors. PMID- 10668174 TI - Synthesis and analgesic profile of novel N-containing heterocycle derivatives: arylidene 3-phenyl-1,2,4-oxadiazole-5-carbohydrazide. AB - This paper describes recent results of a research program aimed at the synthesis and pharmacological evaluation of new heterocyclic N-acylhydrazone (NAH) compounds, belonging to the arylidene (3-phenyl)-1,2,4-oxadiazolyl-5 carboxyhydrazide (8a-p) series. These compounds were structurally planned by applying the molecular hybridization strategy on previously described arylidene 1 phenylpyrazole-4-carbohydrazide (5) derivatives, considered as lead-compounds, which present potent analgesic properties. The analgesic profile of the title compounds 8a-p, evaluated in the model of abdominal constrictions induced by acetic acid, showed that the 4-methoxybenzylidene derivatives 8c and 8k were the most active ones, exhibiting a relative analgesic activity comparable with that of dipyrone 1 used as standard. PMID- 10668175 TI - Formulation of a neutral solution of ciprofloxacin upon complexation with aluminum. AB - Clear solutions of 0.5 and 1.0% ciprofloxacin (CF) of pH 7.2 were prepared by the addition of aluminum chloride hexahydrate (AlCl3.6H2O) in the molar proportion CF:AlCl3.6H2O (3:1). Minimum inhibitory concentrations (MIC) of these solutions were the same as an equimolar solution of CF.HCl. Solutions exhibited good physical, chemical and microbiological stability and satisfactorily overcame an ocular irritation test on rabbits. PMID- 10668176 TI - Synthesis and antimycobacterial activity of some N1-[1-[3-aryl-1-(pyridin-2-, 3-, or 4-yl)-3-oxo]propyl]-2- pyridinecarboxamidrazones. AB - N1-[1-[3-aryl-1-(pyridin-2,3-, and 4-yl)-3-oxo[propyl]-2- pyridinecarboxamidrazone derivatives were synthesized and tested for their in vitro antimycobacterial activity. Some compounds showed interesting activity against a strain of Mycobacterium tuberculosis and a strain of Mycobacterium avium. PMID- 10668177 TI - Synthesis and antinociceptive activity of (1-benzyl-2(3H)-benzimidazolon- 3-yl) acetic acid derivatives. AB - Eight (1-benzyl-2(3H)-benzimidazolon-3-yl)acetic acid derivatives have been synthesized and tested for antinociceptive activity in this study. All compounds but one, at the oral dose of 100 mg/kg were comparable with aspirin. Ethyl (1 benzyl-2(3H)-benzimidazolon-3-yl)acetate (3), 4-[(1-benzyl-2(3H)-benzimidazolon-3 yl)acetyl]morpholine (6a) and 1-[(1-benzyl-2(3H)-benzimidazolon-3 yl)acetyl]pyrrolidine (6b) have shown more potent antinociceptive activity than others. PMID- 10668178 TI - Investigations on the synthesis and pharmacological properties of amides of 7 methyl-3-phenyl-1-[2-hydroxy-3-(4-phenyl-1-piperazinyl)propyl]-2,4- dioxo-1,2,3,4 tetrahydropyrido[2,3-d]-pyrimidine-5-carboxylic acid. AB - Synthesis of amides of 7-methyl-3-phenyl-2,4-dioxo-1,2,3,4- tetrahydropyrido[2,3 d]pyrimidine-5-carboxylic acid (6-10) and their 1-[2-hydroxy-3(4-phenyl-1 piperazinyl)propyl] derivatives (11-15) are described. Some of them displayed strong analgesic activity. PMID- 10668179 TI - 4-quinazolinones: synthesis and reduction of prostaglandin E2 production. AB - We synthesized and evaluated the anti-inflammatory activity of a series of 4 quinazolinone derivatives. Two approaches were used to yield the title compounds. A first group of quinazolinone derivatives was obtained by the appropriate substituted anthranilates. A second group of quinazolinone compounds was prepared through the benzoxazin-4-ones intermediate. The pharmacological results reveal that the synthesized derivatives exhibit a significant anti-inflammatory effect in an experimental ocular inflammation model. In fact, all the tested compounds lowered the prostaglandin E2 (PGE2) production with respect to the control group (P < 0.05). The 3-cyclohexyl-6-chloro-quinazolin-4(3H)-one and 3-cyclohexyl quinazolin-4(3H)-one derivatives were the most active compounds. These compounds significantly reduced PGE2 levels even more than the reference drug tolmetin and significantly lower protein concentration and polymorphonuclear leukocytes number compared to the control group (P < 0.05). Therefore, these compounds may be useful to prevent ocular inflammatory reactions. PMID- 10668180 TI - Synthesis and biological activity of pseudopeptides inhibitors of Ras farnesyl transferase containing unconventional amino acids. AB - A study was performed on the structure-activity relationships of a series of phenol derivatives, CVFM analogs, derived from the two most active compounds of a first series (1A and 1B) of inhibitors of Ras farnesyl transferase (FTase) that we have recently described. We report the synthesis and the activity of a second series of compounds in which the phenylalanine residue was replaced by unconventional aromatic and non-aromatic amino acids, with varying electronic, lipophilic, steric and conformational properties. The compounds showed to be significantly less active than reference compounds against FT, with the only exception of derivative 3A (IC50 = 3 microM), which is slightly more active than 1A but not 1B. Subsequently we tested the effects of compounds 1A, 1B and 3A, 3B on the anchorage-dependent growth of two epithelial cell lines of rats, FRTL-5 and the same line v-Ha-ras transformed. Compound 3A derived from lead compound 1A, showed an appreciable selectivity against transformed cells. In contrast, compounds derived from derivative 1B had only a modest cellular activity. PMID- 10668181 TI - Synthesis of various analog derivatives of ORG13514 as 5-HT1A ligands. AB - In connection with the development of new potential 5-HT1A ligands, multistep synthesis of N-substituted-3-aminomethyl-2,3-dihydro-1,4-dioxinol[2,3-b]pyridin e derivatives as ORG13514 analogs are described. Their biological activity as 5 HT1A type ligands is reported and compared with ORG13514 affinity and selectivity for 5-HT1A receptors. PMID- 10668182 TI - Sterically controlled regiospecific heterocyclization of 3-hydrazino-5-methyl 1,2,4-triazino[5,6-b]indole to 10-methyl-1,2,4-triazolo[4',3':2,3[1,2,4 triazino[5,6-b]indoles. AB - 3-Hydrazino-5-methyl-1,2,4-triazino[5,6-b]indole underwent sterically controlled regiospecific heterocyclizations with a variety of one-carbon cyclizing agents to give the sterically more favored linearly annulated 10-methyl-1,2,4 triazolo[4',3':2,3[1,2,4-triazino[5,6-b]indoles rather than the sterically less favored angularly annulated 10-methyl-1,2,4-triazolo[3',4':3,4]1,2,4-triazino[5,6 b]indoles. The assigned structures were corroborated by comparison with unequivocally synthesized authentics, chemical and spectral data. The antimicrobial activity of some of the prepared compounds was investigated. PMID- 10668183 TI - Structure-activity studies on nociceptin/orphanin FQ: from full agonist, to partial agonist, to pure antagonist. AB - A heptadecapeptide (Phe-Gly-Gly-Phe-Thr-Gly-Ala-Arg-Lys-Ser-Ala-Arg-Lys-Leu-Ala Asn-Gln) was identified from rat brain and from porcine brain as a ligand for OP4, a new G-protein coupled receptor that is similar in sequence to opioid receptors. The OP4 receptor is widely expressed in the nervous system where it mediates a broad range of physiological functions. The new peptide, nociceptin (NC), has a primary sequence recalling that of opioid peptides. Despite the homologies (a) of the OP4 receptor with known opioid receptors, especially the OP2 (kappa) receptor, and (b) of NC with opioid peptides, particularly dynorphin A, the two biological systems have different anatomical locations and chemical requirements for activation. NC does not bind to opioid receptors, and mammalian opioid peptides do not interact with the OP4 receptor. The presence of Phe in position 1 and Arg in position 8, appear to be instrumental to exclude NC from interacting with the opioid receptors. Contrary to opioid peptides which strikly require Tyr in position 1, the active core that activates the OP4 appears to be towards the centre of the peptide molecule and includes Phe4. Based on the message/address model, several changes have been made in the N-terminal tetrapeptide Phe-Gly-Gly-Phe (message) and a few also in the C-terminal of the template NC(1-13)-NH2, a fragment that acts as a full agonist both in vitro and in vivo. Subtle changes of the N-terminal sequence, especially at Phe1, led to the discovery of peptide antagonists ([Phe1 psi (CH2-NH)Gly2[-NC(1-13)-NH2 and [Nphe1[-NC(1-13)-NH2). The first compound has been widely used to characterize NC actions in the periphery and in the central nervous system. It has been shown to act mainly as an antagonist outside the brain and as an agonist in the central nervous system. [Nphe1[-NC(1-13)-NH2- on the contrary, acts as antagonist both in the periphery and in the brain. These first peptide prototypes may soon be followed by non-peptide compounds, some of which, are already described in patient literature. PMID- 10668184 TI - 5-Nitroimidazole derivatives as possible antibacterial and antifungal agents. AB - Some novel 1-[2-[[5-(2-furanyl)-4-substituted 4H-1,2,4-triazol-3-yl[thio[ethyl[-2 methyl-5-nitro-1H-imidazoles (3), 1-[3-[[5-(2-furanyl/2-thienyl)-4-substituted 4H 1,2,4-triazol-3-yl[-thio]-2-hydroxypropyl[-2-methyl-5-nitro-1H- imidazoles (5) and 1-[3-[(N,N-disubstituted thiocarbamoyl)-thio[-2-hydroxypropyl]-2-methyl-5 nitro-1H-imidazoles (7) were synthesized and evaluated for in vitro antibacterial and antifungal activity. Some of 5 were found to be effective against bacteria and fungi (minimum inhibitory concentration (MIC) 7.3-125 micrograms/ml), whereas 7 were found to be effective against fungi (MIC 3-25 micrograms/ml). PMID- 10668185 TI - Bioavailability of dalarelin--a superactive GnRH analogue--in rats. AB - 125I-marked dalarelin (a modified analogue of GnRH) or GnRH (gonadotropin releasing hormone) were administered to the tested rats in single doses of 127 ng/kg by subcutaneous injections. Dalarelin and GnRH were absorbed from the injected doses in 0.64 and 0.49%, respectively. Only one remarkable maximal concentration of these hormones was noticed in rats' blood 30 min after the administration. Dalarelin maximal concentration was 261.5 pg/cm3 and was 93.43% higher than the maximal concentration of GnRH. Dalarelin bioavailability was 1651.89 pg/cm3, whereas GnRH bioavailability was 718 pg/cm3 h. The bioavailability level of dalarelin was 230% compared with that for GnRH, which was accepted as a pattern of bioavailability. PMID- 10668186 TI - Anticonvulsant activity of analogues of acetazolamide. AB - The anticonvulsant activity of 5-tertbutyloxycarbonylamido-1,3,4-thiadiazole-2 sulfonamide (B-H2ats) and 5-amino-1,3,4-thiadiazole-2-sulfonamide (Hats) was compared in mice, to that of acetazolamide (H2acm). These compounds exhibit potent anticonvulsant activity and low minimal motor impairment. PMID- 10668187 TI - Synthesis and anticonvulsant activity of some (2/4-substituted)benzaldehyde (2 oxobenzothiazolin-3-yl)acetohydrazones. AB - Fifteen new (2/4-substituted)benzaldehyde (2-oxobenzothiazolin-3 yl)acetohydrazones were synthesized and their structures were elucidated by NMR and elemental analysis. Their anticonvulsant activity was tested by a pentylenetetrazole induced seizure test. Compounds 4e and 4h were found to be the most promising among the others. PMID- 10668188 TI - Synthesis and antinociceptive activity of some 3-substituted benzothiazolone derivatives. AB - Thirteen 3-substituted benzothiazolone derivatives have been synthesized. Their chemical structures have been elucidated by IR and NMR spectral data and by elemental analyses. Among these compounds, 1-?3-[2(3H)-benzothiazolon-3 yl[propanoyl]morpholine (5b); 1-?3-[2(3H)-benzothiazolon-3-yl[propanoyl]-4 benzylpiperidine++ + (5c); 1-?3-[2(3H)-benzothiazolon-3-yl[-propanoyl]-4 phenylpiperazine (5d); 3-[3-(4-benzylpiperidine-1-yl)propyl]-2(3H) benzothiazolone (5k); 3-[3-(4-benzylpiperazine-1-yl)propyl]-2(3H)-benzothiazolone (5I); 3-[3-(4-phenylpiperazine-1-yl)propyl]-2(3H)-benzothiazolone (5m) have been found to be significantly more active than the others. PMID- 10668189 TI - Intermediate filament protein expression and sugar moieties in normal canine placenta. AB - In the female dog, the placenta is considered zonal, endotheliochorial and labyrinthic. The distribution of the intermediate filaments as well as the surface glycoproteins in the canine placenta are still unknown. The aim of the present study was to provide this information for further understanding of pathological conditions in the bitch. Samples were obtained from normal uterine horns at the end of gestation. Tissues were routinely fixed and stained. Monoclonal antibodies against cytokeratins, vimentin and desmin were used for immunohistochemical staining. UEA-1, PNA, RCA-1, SBA, DBA, WGA and ConA were used for the lectin histochemical staining. A computer morphometrical analysis was made. Statistical analysis was then accomplished. The results showed the maximum immunohistochemical percentage for vimentin in the supraglandular connective tissue, for pancytokeratin in the spongy zone and for desmin in miometrium. SBA showed the highest staining percentage in the gland cells of the spongy zone, while ConA was the highest in the syncytiotrophoblastic cells and gland cells of the deep glandular zone. The results obtained indicate that the lectin binding pattern is partially different from other animal species. On the contrary, the intermediate filament data coincide with similar observations from other mammals. PMID- 10668190 TI - Effect of irradiation on autogenous bone transplantation in rat parietal bone. AB - To determine the appropriate time for bone reconstruction after irradiation, the healing process after autogenous iliac bone transplantation in the irradiated parietal bone was examined by scanning electron microscopy and light microscopy. Bone transplantation was carried out at the second and the fourth weeks after Cobalt-sixty (60Co) irradiation with calculated dose and fractionation. Animals without irradiation were used as control. The results show the appearance of mesenchymal cells and blood vessels around the transplantation to be extremely few one week after transplantation which was carried out at the second week after irradiation. These inhibitions were still seen two weeks after transplantation. Four weeks after transplantation, there were no differences in the bone formation among the experimental groups. Bone formation in the transplantation at the fourth week after irradiation was similar to that of the control group. Microvascularization in the transplantation at the second week after irradiation was inhibited one week after transplantation. The delay in bone healing was responsible for the retardation of revascularization and caused microcirculatory failures as well as the damage of osteogenic cells. It is quite clear that damaged cells and tissues recovered by the elapse of time under the irradiation procedure employed in this study and also that bone formation was carried out in the physiological process. We think that bone transplantation after irradiation should be done after recovery from the radiation damage to the periosteal cells and the blood vessels. PMID- 10668191 TI - Functional status of the immune system after chronic administration of 2' deoxycoformycin in the BB rat. AB - Insulin-dependent diabetes mellitus (IDDM) is caused by autoimmune destruction of pancreatic beta cells with the primary mechanism being cell mediated. The BB rat develops insulitis and IDDM with many features analogous to the disease in man. In previous studies we reported that weekly administration of 2'-deoxycoformycin (dCF) for four months reduces significantly the incidence of IDDM in the BB rat by 70%, and that the animals remain free of diabetes for a minimum of two months after drug withdrawal. Since the diabetes-prone BB rat is lymphopenic, with a reduction of both CD4 and CD8 cells, the continuous failure of dCF treated animals to develop diabetes may have been due to generalized immunosuppression. To test this possibility, the ability of dCF treated diabetes-free BB rats to mount an immune response after challenge with Ovalbumin was examined five months after drug withdrawal. The results showed that the post-immunization levels of total IgG and specific IgG in these animals did not differ from those observed in non-dCF treated controls nor those of control diabetes-resistant non-lymphopenic BB rats. Moreover, FACS analysis indicated no change in the percentages of total numbers of CD4+ or CD8+ cells between the two groups of animals. Histological assessment of the pancreata of the post-dCF treated animals showed varying degrees of mononuclear cell infiltrates in the islets. These data demonstrate that treatment by dCF is not permanent, and may require intermittent or continuous administration to prevent development of diabetes. Further studies are needed to determine the mechanism of action of dCF in this model of IDDM. PMID- 10668192 TI - Studies on the breakdown of glycogen in the lysosomes: the effects of hydrocortisone. AB - The effects of hydrocortisone on newborn rat liver were studied by using biochemical assays, electron microscopy and quantitative morphometry. Hydrocortisone increased the number of lysosomes in the hepatocytes. Most of the lysosomes represented glycogen-containing autophagic vacuoles. The glucocorticoid also increased the activity of the liver glycogen-hydrolyzing acid glucosidase and the breakdown of glycogen inside lysosomes. The activity of the liver acid mannose 6-phosphatase was decreased. This may be related to the stimulation of autophagic mechanisms in the newborn rat hepatocytes. PMID- 10668193 TI - Does diabetic state affect co-localization of peptide YY and enteroglucagon in colonic endocrine cells? AB - BACKGROUND: Changes in the numbers of PYY- and enteroglucagon-immunoreactive cells in colon of animal models of human diabetes have been reported. As these peptides co-localize in the same cells it is possible that the observed changes are a result of changes in co-localization. METHODS: Animal models of human type 1 and type 2 diabetes, namely the non-obese diabetic (NOD) mouse and the obese (ob/ob) mouse, were studied. As controls for the NOD mice, BALB/cJ mice were used and for ob/ob mice, homozygous lean (+/+) mice were used. Tissue samples from colon were double-immunostained for PYY and enteroglucagon according to the indirect immunofluorescence method. RESULTS: Co-localization of enteroglucagon and PYY was found in colonic endocrine cells in all groups investigated. Compared with controls, pre-diabetic NOD mice showed a decreased proportion of enteroglucagon/PYY co-localization. There was no difference in diabetic NOD mice or diabetic ob/ob mice when compared with controls. CONCLUSIONS: Whereas the number of cells containing solely enteroglucagon and solely PYY increases in pre diabetic NOD mice, production of enteroglucagon in PYY-immunoreactive cells decreases. Although the numbers of PYY and enteroglucagon cells have been reported to be changed in both diabetic NOD mice and in obese mice, the balance between co-expressing and mono-expressing cells seems to be preserved. PMID- 10668194 TI - Topological differences along mammalian motor nerve terminals for spontaneous and alpha-bungarotoxin-induced sprouting. AB - Spontaneous sproutings can be observed in end plates from normal adult vertebrate muscles and motor end plates develop increased growth signs and sprouts when target muscle cells become less active or paralysed. Nevertheless, very little is known about where in the motor nerve terminal arborization spontaneous and experimentally induced sprouts originate, their similarities and differences and also about their final maturation or elimination. In this study we investigate the topological properties of both spontaneous and alpha-bungarotoxin-induced sprouts (during different periods of intoxication and after recovery) along the motor nerve terminal branches of the Levator auris longus muscle of Swiss mice (between 48-169 day old). Muscles were processed for immunocytochemistry to simultaneously detect postsynaptic AChRs and axons. This procedure permits us to make an accurate identification of the fine sprouts and a morphometric study of the presynaptic branching pattern profile in control muscles, during the toxin action and after recovery from paralysis. The results show that in normal muscles, the initial and trunk segments (those between branch points) of the terminal arborization sprouted proportionally more branches when taking their relative lengths into account than the distal free-end segments. In contrast, every micrometer of alpha-bungarotoxin-treated muscles throughout the full terminal arborization have the same probability of generating a sprout. Moreover, the toxin-induced sprouts can consolidate as new branches once recovered from the paralysis without changing the total length of the nerve terminal arborization. PMID- 10668195 TI - Apoptosis in gallbladder carcinomas and dysplasias, its relation to the expression of caspases 3, 6 and 8 and apoptosis regulating proteins bcl-2, mcl-1 and bax. AB - In this study we investigated apoptosis and the expression of caspases 3, 6 and 8 and bcl-2, mcl-1 and bax in 39 gallbladder carcinomas and 7 epithelial dysplasias. The average apoptotic index was 0.68 +/- 0.91%. The extent of apoptosis was higher in grade II-III than grade I tumours or epithelial dysplasias (p = 0.003). Also, tumours invading beyond serosa or into other organs (T3-T4) had a higher apoptotic index than other tumours (p = 0.05). Caspase 3 expression was found in 37 (95%) and caspase 6 and 8 expression each in 30 (77%) carcinomas. Their expression associated with each other and tended to increase along with the progression of the lesions. Bcl-2 expression was found in only 4 (10%) tumours. In contrast, mcl-1 positivity was found in 34 (87%) and bax positivity in all cases. The results show that apoptosis is increased along with progression of the neoplastic lesions of the gallbladder epithelium. Caspases 3, 6 and 8 are strongly expressed in gallbladder carcinomas suggesting that they contribute to the increased apoptosis observed in them. Of the bcl-2 family proteins, bcl-2 was expressed infrequently suggesting that it does not play any significant role in apoptosis inhibition in gallbladder tumours. PMID- 10668196 TI - Aging affects different human muscles in various ways. An image analysis of the histomorphometric characteristics of fiber types in human masseter and vastus lateralis muscles from young adults and the very old. AB - This study is an attempt to objectively evaluate age-related changes in human muscles by use of histomorphometric methods. Aging in humans induces dramatic transformations in the skeletal muscles but little is known as to whether or not the aging processes per se may affect all muscles equally. In this study aging of two human muscles with different functions, origin and nerve supply is compared. Sections were cut from masseter and vastus lateralis muscles obtained from young adults aged 18-24 years and from the very old aged 90-102 years. Muscle fiber types were classified with the traditional myofibrillar ATPase staining. Various histomorphometric parameters of the different fiber types in human masseter and vastus lateralis muscle sections were obtained by image analyses to evaluate the age-related changes in the muscle fibers. The following variables were calculated: the number of each fiber type per photographed area; the area of each fiber and two indicators for the shape of the muscle fibers. In the aging muscles there was no relative preferential loss of a fiber type. High numbers of intermediate ATPase-stained fibers (IM fibers) were found in some old vastus muscles but were only sporadic in young vastus muscles. However, there was no change in the percentage distribution of intermediate ATPase-stained fibers when young and very old human masseter muscles were compared. Incubation of the sections with antimyosin antibodies showed that the IM fibers in old masseter and old vastus contained different myosin heavy chains. Thus ATPase activity and anti myosin staining displayed a somewhat different pattern of fiber type distribution. The main changes in the shape and area indicated that type I fibers in the masseter became more circular while in the vastus they decreased significantly in size. The type II fibers in the vastus became very small and deviated significantly from circularity whereas the type II fibers in the masseter only exhibited a decrease in the size of the fibers. Histomorphometric measurements show that aging affects different human muscles in various ways. PMID- 10668197 TI - Effect of hypertension on the angiotensin II fibres arriving at the posterior lobe of the hypophysis of the rat. An immunohistochemical study. AB - We studied immunohistochemically the posterior lobe of the hypophysis (PL) of 15 week-old spontaneously hypertensive rats (SHR) and of matched normotensive Wistar Kyoto rats (WKY), by using our own polyclonal antibody raised in mice against Angiotensin II (mouse-antiangiotensin II, MAAII). The blood pressure, water intake and volume of the PL were also recorded. The SHR rats were hypertensive, drank more water and showed a clear hypertrophy of their hypophysial PL. Also the PL of the SHR animals showed an increase in the immunoreactivity to the anti angiotensin II antibody in the fibres arriving at the PL, with respect to the PL of WKY rats. This increase is compatible with the hyperactivity of the brain RAS, depletion of vasopressin content in the PL and increase in plasmatic levels of vasopressin described in SHR rats with respect to normotensive animals, as angiotensin II could locally stimulate vasopressin release to plasma from the neurohypophysis. PMID- 10668198 TI - Human papillomaviruses and DNA ploidy in anal condylomata acuminata. AB - Previous studies have emphasized the usefulness of DNA ploidy measurement and Human Papillomavirus (HPV) detection as prognostic markers in low grade cervical lesions. We addressed the eventual relationship between HPV type, DNA profile, and p53 tumor suppressor protein expression in anal condylomata acuminata to eventually determine parameters which may be considered as predictive risk factors for the development of cancer. DNA ploidy was assessed by image cytometry after Feulgen staining of contiguous serial sections of 45 anal condylomata acuminata without atypia containing HPV detected by in situ hybridization and Polymerase Chain Reaction (PCR). p53 expression was detected by immunohistochemistry. DNA aneuploidy was found in 53.3% of these lesions, 48.9% containing non oncogenic HPV types 6 and/or 11 and 4.4% harbouring HPV types 11 and 18. The DNA diploid lesions were all associated with non oncogenic HPV types 6 and/or 11 and one case also contained HPV type 33. There was no significant correlation between the detection of DNA aneuploidy and the presence of immuno detected p53. DNA aneuploidy was not related to the presence of oncogenic HPV in anal condylomata acuminata. The DNA aneuploid profile frequently observed, especially in lesions associated with non oncogenic HPV types, is not yet well explained and cannot be considered as a prognostic factor. In contrast, a more intensive clinical follow-up should be proposed in patients with oncogenic HPV associated to DNA aneuploidy. PMID- 10668199 TI - Expression of galectin-1 and -3 and of accessible binding sites during murine hair cycle. AB - Although protein-carbohydrate interactions are supposed to play key roles in cell adhesion, signalling and growth control. Their exact role in skin physiology has only recently been investigated. The endogenous lectins galectin-1 and galectin-3 have been identified in skin including hair follicles. Here, we analyzed the expression and distribution of these galectins and their binding sites in C57BL/6 mice during hair cycle. The expression of galectin-1 and galectin-3 binding sites was found to be predominantly hair cycle-dependent showing some overlapping to the expression of galectin-1 and -3. The outer root sheath (ORS) expressed galectin-1 binding sites during anagen IV to VI and in early catagen, whereas galectin-1 was expressed from early anagen to late catagen. The ORS expressed galectin-3 binding sites during catagen transition corresponding to a galectin-3 expression during anagen V and catagen. The innermost layer of the ORS expressed galectin-3 binding sites during anagen VI until catagen VIII, but galectin-3 during anagen III to IV and catagen. The inner root sheath (IRS) expressed galectin-3 binding sites only in anagen IV but missed expression of any of the two galectins. The matrix cells expressed galectin-3 binding sites in catagen II III as well as galectin-3 during anagen V to catagen IV. The present study provides the first evidence for a cycle-related expression of both galectin-1 and -3 and their binding sites during murine hair cycle. PMID- 10668200 TI - Elastin variations implicating in vascular smooth muscle cells phenotype in human tortuous arteries. AB - The aim of the present work was to study the morphological implications between the elastin and the phenotypic expression of the vascular smooth muscle cells. For this purpose, sixty human tortuous arteries from different territories have been studied. We have measured the morphometric indexes Intimal Thickening Index and Elastolyse Index and they have been quantified with computer system analysis, image-colour corresponding to the orcein and Verhoeff reactions for detecting elastin and the alpha-actin in the smooth muscle cells. We compared both territorial arteries from the cranial and from abdominal origin. The elastin concentration was similar in both territories, but not its morphology according to its spatial distribution. We have observed a relationship between the elastin structural organisation from the media of arteries and of the internal elastic lamina in these territories and the variation of reactivity to the smooth muscle alpha-actin as a marker of the phenotypic state. Our results confirm the hypothesis that elastin, besides intervening in the architecture of the arterial wall, is a factor implicated in the phenotypic variability of the smooth muscle cells and in the development and evolution of the intimal thickenings in human atherosclerosis. PMID- 10668201 TI - The molecular determinants of the efficiency of green fluorescent protein mutants. AB - The Green Fluorescent Protein (GFP) is a spontaneously fluorescent polypeptide of 27 kD from the jellyfish Aequorea victoria that absorbs UV-blue light and emits in the green region of the spectrum. GFP has been successfully expressed both in bacteria and in eukaryotic cells and is widely used to monitor the localization of tagged proteins in living cells. Since wtGFP performs inefficiently in different cellular contexts, efforts have been devoted to the improvement of GFP expression levels and/or fluorescence. We will here review the basic characteristics of wt and mutated GFP, in particular their protein expression vs fluorescent properties. Emphasis will be given to unexpected consequences of mutations of the GFP gene, i.e. on transcription and translation rates and on protein folding in different cell types, and to how these critically reflect on the use of GFP in different cellular environments. PMID- 10668202 TI - The yin-yang of PR-domain family genes in tumorigenesis. AB - Cancer is essentially caused by alterations in normal cellular genes. Multiple gene changes involving at least two types of cancer genes, protooncogenes and tumor suppressor genes, are required for the clonal expansion of a malignant cell. This discussion focuses on the recently recognized role of a small but expanding family of PR-domain genes in tumorigenesis. The protein products of these genes are involved in human cancers in an unusual yin-yang fashion. Two products are normally produced from a PR-domain family member which differ by the presence or absence of the PR domain; the PR-plus product is disrupted or underexpressed whereas the PR-minus product is present or overexpressed in cancer cells. This imbalance in the amount of the two products, a result of either genetic or epigenetic events, appears to be an important cause of malignancy. PMID- 10668203 TI - The role of the epidermal growth factor-like protein dlk in cell differentiation. AB - This review focuses on the current knowledge about the function of the EGF-like homeotic protein dlk. dlk is a transmembrane protein that possesses six Epidermal Growth Factor-like sequences at the extracellular domain, a single transmembrane domain and a short intracellular tail. Because of its overall structure and amino acid homology, dlk belongs to the EGF-like homeotic protein family. This family includes proteins such as the Notch receptor and its homologues, as well as Notch ligands, such as Delta, Serrate, and their mammalian homologues Dll1, Dll2 and Dll3 and Jagged 1 and Jagged 2. (For a recent review see Fleming, 1998). dlk is highly expressed by preadipose cell lines, and neuroendocrine tumors, such as pheochromocytomas and neuroblastomas. dlk has been involved in several differentiation processes, such as adipogenesis, hematopoiesis and B cell lymphopoiesis, and neuroendocrine differentiation, including the differentiation of pancreas and the adrenal gland. The extracellular region of dlk can be released by action of an unknown protease and this soluble dlk variant accumulates in the amniotic fluid and is able to inhibit adipocyte differentiation in vitro. Recent evidence indicates, however, that membrane associated dlk variants play a positive role in the differentiation process. These findings suggest that dlk plays an important role in differentiation and tumorigenesis of several cellular types. PMID- 10668204 TI - The female prostate and prostate-specific antigen. Immunohistochemical localization, implications of this prostate marker in women and reasons for using the term "prostate" in the human female. AB - Prostate-specific antigen (PSA) is currently the most frequently used marker for the identification of normal and pathologically altered prostatic tissue in the male and female. Immunohistochemically PSA is expressed in the highly specialized apically-superficial layer of female and male secretory cells of the prostate gland, and as well as in uroepithelial cells at other sites of the urogenital tract of both sexes. Unique active moieties of cells of the female and the male prostate gland and in other parts of the urogenital tract are indicative of secretory and protective function of specialized prostatic and uroepithelial cells with strong immunological properties given by the presence of PSA. In clinical practice, PSA is a valuable marker for the diagnosis and monitoring of diseases of the male and the female prostate, especially carcinoma. In the female, similarly as in the male, the prostate (Skene's gland) is the principal source of PSA. The value of PSA in women increases in the pathological female prostate, e.g., carcinoma. Nevertheless, the total amount of PSA in the female is the sum of normal or pathological female prostate and non-prostatic female tissues production, e.g., of diseased female breast tissue. The expression of an antigen specific for the male prostate, i.e., PSA in female Skene's glands and ducts, and structural and functional parameters and diseases similar to that of the male prostate, have provided convincing evidence of the existence of a prostate in women and definitive preference of the term "prostate" over that of Skene's glands and ducts. The use of the term Skene's glands incorrectly implies that some other structure rather than prostate is involved, promoting the vestigial position of this female organ. PMID- 10668205 TI - The involvement of Helix pomatia lectin (HPA) binding N-acetylgalactosamine glycans in cancer progression. AB - The lectin from Helix pomatia, the Roman snail (HPA), recognises terminal alpha N acetylgalactosamine residues. A large number of lectin histochemical studies have demonstrated that expression of HPA-binding glycoproteins by cancer cells to be a marker of metastatic competence and poor prognosis in a range of common human adenocarcinomas, including those of breast, stomach, ovary, oesophagus, colorectum, thyroid and prostate. Around 80% of metastases arising from primary breast cancer are predictably HPA positive, but, intriguingly, around 20% do not express HPA binding glycoproteins reflecting the complexity of metastatic mechanisms and the further disruptions in cellular glycosylation that attend tumour progression. HPA binding is not an independent prognostic factor, but is strongly associated with the presence of metastases in local lymph nodes. It does appear to be independent of other clinical features of prognostic importance such as tumour size, histological grade, S-phase fraction, ploidy, and there is little convincing evidence of any association with oncogene expression or hormone receptor positivity. The precise nature of the metastasis-associated HPA binding partner(s) is a question of some interest, but thus far remains unclear. HPA will recognise, for example, the Tn epitope and blood group A antigen, but its prognostic significance appears to be through recognition of a much broader and heterogeneous array of N-galactosaminylated glycoproteins. Their synthesis appears to be mediated through alteration in expression or activity of one or more of the enzymes of glycosylation. The most likely putative roles of HPA binding ligands in the metastatic cascade may be enhancement of invasive capacity, or interaction with an as yet unidentified lectin-like receptor facilitating adhesion processes. The prognostic information provided by HPA lectin histochemistry may be used clinically to inform the physician and aid treatment decisions; far more interesting is the challenge of further understanding the precise nature of the HPA-binding ligands, and defining their role in the complex mechanisms of metastasis. PMID- 10668206 TI - Gene expression and cell turnover in human renal dysplasia. AB - Kidney malformations are common causes of chronic renal failure in children. Dysplastic kidneys represent a unique model of perturbed epithelial-mesenchymal interaction which leads to the formation of malformed branching tubules surrounded by undifferentiated and metaplastic mesenchymal cells. We have found that human dysplastic epithelia express PAX2 (a transcription factor), BCL2 (a survival factor) and galectin-3 (a cell adhesion/signaling molecule). These genes are implicated in oncogenesis and their persistent expression may drive proliferation of dysplastic cysts, hence explaining the massive growth of some multicystic dysplastic kidneys. We have also detected prominent apoptosis in undifferentiated tissues around dysplastic epithelia, and this may provide a potential mechanism for the well-documented regression of dysplastic kidneys. Hence, although these kidneys may not have any excretory function, it is incorrect to consider them as 'end stage organs' because they are highly active in terms of cell turnover and gene expression; furthermore, these processes can be correlated with patterns of tissue growth and involution. Further elucidation of 'molecular lesions' in renal malformations may lead to novel therapies to enhance the differentiation of progenitor cells. PMID- 10668207 TI - The role of activated cytotoxic T cells in inflammatory bowel disease. AB - The role of cell-mediated cytotoxicity in the pathogenesis of ulcerative colitis and Crohn's disease has been controversial since reports indicating either a decreased, or an increased, activity of cytotoxic T cells in active stages of inflammatory bowel disease exist. Some of these discrepancies may be attributed to the fact that so far mostly peripheral blood lymphocytes rather than intestinal T cells have been examined. To overcome some of these limitations we performed in situ hybridizations for the detection of perforin and granzyme A mRNA expressing cells, i.e. of cytotoxic cells activated in situ, in the affected intestinal mucosa. These studies revealed increased frequencies of activated, cytotoxic T cells in active stages of ulcerative colitis and Crohn's disease. Interestingly, activated perforin mRNA expressing T cells are present both in the CD4 and in the CD8 T cell subsets. In the latter T cell subset up to 60% of the mucosal T cells isolated from the affected sites express perforin mRNA at detectable levels. The elevated frequency of activated cytotoxic cells and their histological distribution also in close proximity to the epithelial cells may thus indicate an important role for cytotoxic cells in the pathogenesis of inflammatory bowel disease since activated cytotoxic T cells may further exacerbate the inflammatory process through the production of pro-inflammatory cytokines such as interferon-gamma or tumor necrosis factor-alpha, but also through the release of pro-inflammatory cytokines and chemokines upon lysis of epithelial cells and the increased influx of luminal antigens at the site of epithelial erosions. PMID- 10668208 TI - The visualization of oxidant stress in tissues and isolated cells. AB - Many studies have implicated the role of oxidant stress in a wide range of human diseases and have led to the rapid expansion of research in this area. With many experimental approaches a direct detection of the production of reactive oxygen species (ROS) and free radicals is not possible. Free radicals are very reactive, short-lived and react in a non-specific way, so that ongoing oxidative damage is generally analyzed by measurement of secondary products e.g. H2O2, "oxidized" proteins, peroxidized lipids and their break-down products, "oxidized" DNA or by fluorographic analysis in combination with fluorescent dyes e.g. dichlorofluorescin (DCFH). The histochemical visualization of selected molecular markers for oxidative phenomena can often provide valuable information concerning the distribution of oxidative processes in vivo. A number of biochemical methods are available for the monitoring of almost all oxidant stress-related processes, although their applicability in vivo is limited. This review summarizes the biochemical methods currently available for histochemical detection and indirect visualization of an excess of free radicals and ROS. The cited methods are discussed and the results obtained from their application are critically evaluated. PMID- 10668209 TI - Organ specificity of the structural organization and fine distribution of lymphatic capillary networks: histochemical study. AB - Histochemical studies of the microcirculatory system were reviewed with regard to the organ specificity of the structural organization and fine distribution of the lymphatic capillary network. The lymphatics and blood vessels are characterized by an enzyme-histochemical method using 5'-nucleotidase (5'-Nase), alkaline phosphatase (ALPase) and/or diaminopeptidase (DAPase) staining in addition to an immunohistochemical method. The 5'-Nase-positive lymphatic vessels can be distinguished histochemically from arterial and venous vessels based on ALPase and DAPase activity, respectively. The specificity and localization of the enzyme reactions were confirmed by comparative histochemical studies of the same specimen with light microscopy and scanning or transmission electron microscopy. These histochemical methods are discussed in relation to their ability to demonstrate the organ specificity of vascular networks under normal and pathological conditions. PMID- 10668210 TI - Current understanding of macrophage type 1 cytokine responses during intracellular infections. AB - Macrophages are important effector cells in cell-mediated immunity against intracellular infection. Among cytokines that macrophages are able to release are IL-12 and TNF alpha. IL-12 is a critical linker between the innate and adaptive cell-mediated immunity, capable of Th1 differentiation and IFN gamma release by T and NK cells. IFN gamma is critically required for the activation of macrophage bactericidal activities. Recently emerging evidence suggests that macrophages are able to release not only IL-12 and TNF alpha but also IFN gamma. However, the mechanisms that control the release of each of these type 1 cytokines in macrophages appear different. While macrophages release TNF alpha in an indiscriminate and IL-12-independent way, the release of IL-12, particularly bioactive IL-12 p70, and IFN gamma is under tight control. We are just beginning to understand what controls the release of IL-12 p70, a question of fundamental importance to understanding the mechanisms underlying the initiation of cell mediated immunity. Our recent findings have shed more insights into the regulatory mechanisms of macrophage IFN gamma responses. It has become evident that IL-12 is required not only for Th1 differentiation but also for IFN gamma responses by both T cells and macrophages during intracellular infection. In this overview, we have discussed about the current understanding of the regulation of macrophage type 1 cytokine responses during intracellular infection, based upon the recent findings from us and others. PMID- 10668211 TI - Histopathology of the male reproductive system induced by the fungicide benomyl. AB - Benomyl is an effective fungicide that has been in use for many years. This chemical and its primary metabolite, carbendazim, are microtubule poisons that are relatively nontoxic to all mammalian organs, except for the male reproductive system. Its primary effects, at moderate to low dosages, are on the testis, where it causes sloughing of germ cells in a stage-dependent manner. Sloughing is caused by the effects of the chemical on microtubules and intermediate filaments of the Sertoli cell. These effects spread to dividing germ cells and also lead to abnormal development of the head of elongating spermatids. At higher dosages, it causes occlusion of the efferent ducts, blocking passage of sperm from the rete testis to epididymis. The mechanism of occlusion appears to be related to fluid reabsorption, sperm stasis, followed by leukocyte chemotaxis, sperm granulomas, fibrosis and often the formation of abnormal microcanals. The occlusion results in a rapid swelling of the testis and ultimately seminiferous tubular atrophy and infertility. In conclusion, studies that reveal long term testicular atrophy following chronic or subchronic exposure to a toxicant should be re-examined for histopathological lesions in the efferent ductules and head of the epididymis. Lesions in the male track that cause blockage may induce permanent testicular damage and a decrease in sperm production. PMID- 10668212 TI - Matrix metalloproteinases in squamous cell carcinoma. AB - Controlled degradation of extracellular matrix (ECM) is essential in many physiological situations including developmental tissue remodeling, angiogenesis, tissue repair, and normal turnover of ECM. In addition, degradation of matrix components is an important feature of tumor growth, invasion, metastasis, and tumor-induced angiogenesis. Matrix metallo-proteinases (MMPs) are a family of zinc-dependent neutral endopeptidases, which are collectively capable of degrading essentially all ECM components. MMPs apparently play an important role in all the above mentioned aspects of tumor development. In addition, there is recent evidence that MMP activity is required for tumor cell survival. At present, several MMP inhibitors are in clinical trials of malignant tumors of different histogenetic origin. In this review we discuss the current view on the role of MMPs and their inhibitors in development and invasion of squamous cell carcinomas, as a basis for prognostication and therapeutic intervention in these tumors. PMID- 10668213 TI - Splicing and the single cell. AB - The selection of alternative splice sites is an important component of cell specific gene regulation in eukaryotic cells. Use of splice sites can be positively and negatively regulated, and often physiologically appropriate splice site choice is achieved by a balance of the two. RNA elements controlling splice site choice are found in both exons and introns, and these determine management by the cellular splicing machinery. However, the molecular basis of how the splicing machinery responds to these signals in different cells is somewhat of a paradox. Thus far the identified proteins which bind to tissue/cell-specific regulatory elements in mammals are expressed in many different tissues, and not just in the regulating tissue. Potential tissue-specific splicing regulators have been identified by non-biochemical means. However, alternative splicing choices are likely to be affected by subtle differences in the splicing machinery in different cells. In this review I suggest that one important factor is the ratio of proteins in different nuclear compartments, which might be established in a cell type specific fashion. PMID- 10668214 TI - Catenins and their associated proteins in colorectal cancer. AB - Colorectal cancer is the second most common cause of cancer mortality in the western world. Colorectal cancer has been well studied, and the genetic steps involved in the adenoma to carcinoma sequence have been well elucidated. The first genetic alteration, found in 85% of adenomas, are mutations in the adenomatous polyposis coli (APC) gene. However, the consequences of this and the exact function of APC in the colon is not fully understood. It has been suggested that APC could function through its regulation of beta-catenin, an ubiquitous cytoskeletal protein with multiple binding specificities resulting in diverse functions including cell growth, adhesion, and migration. Any change in these associations may play a role in colorectal cancer development and progression. PMID- 10668215 TI - Electron microscopic observation of intracellular expression of mRNA and its protein product: technical review on ultrastructural in situ hybridization and its combination with immunohistochemistry. AB - In situ hybridization (ISH) at the electron microscopic (EM) level is essential for elucidating the intracellular distribution and role of mRNA in protein synthesis. Three different approaches have been applied by the investigators in this EM-ISH study: preembedding method; non-embedding method using ultrathin frozen sections; and postembedding method. In order to obtain satisfactory morphological preservation and retain the messages, we routinely utilized 6 microns-thick frozen sections fixed in 4% paraformaldehyde for the preembedding method and tissues embedded in LR White resin for the postembedding method. The hybridization signal intensity by the postembedding method was lower, and non specific signals were relatively frequent, in comparison with the preembedding method. The preembedding method thus appears to be easier and better than the postembedding method from the viewpoint of applicability and preservation of mRNA, although quantitative analysis of the expression of mRNA is rather difficult in the preembedding method. EM-ISH is considered to be an important tool for clarifying the intracellular localization of mRNA and the exact site of specific hormone synthesis on the rough endoplasmic reticulum. The simultaneous visualization of mRNA and encoded protein in the same cells using preembedding EM ISH and subsequent postembedding immunoreaction with protein A colloidal gold complex is also described. This ultrastructural double-staining method for mRNA and encoded protein can be expected to provide an important clue for elucidating the intracellular correlation of mRNA translation and secretion of translated protein. PMID- 10668216 TI - Role of myofibroblasts during normal tissue repair and excessive scarring: interest of their assessment in nephropathies. AB - Following injury, tissue repair process takes place involving inflammation, granulation tissue formation and scar constitution. Granulation tissue develops from the connective tissue surrounding the damaged area and contains vessels, inflammatory cells, fibroblasts and myofibroblasts. Myofibroblasts play an important role in many tissue injuries and fibrocontractive diseases. The process of normal wound repair after tissue injury follows a closely regulated sequence including the activation and the proliferation of fibroblastic cells. In pathological situations, the normal resolution stages are abrogated and the proliferation of myofibroblasts continues, inducing excessive accumulation of extracellular matrix. The differentiation of fibroblastic cells into myofibroblasts is an early event in the development of tissue fibrosis. Myofibroblastic cells express smooth muscle cytoskeletal markers (alpha-smooth muscle actin in particular) and participate actively in the production of extracellular matrix. The evaluation of myofibroblast differentiation in renal biopsies would be useful for histopathologists to appreciate the intensity of tissue injury and particularly to predict the long term outcome of some nephropathies. Immunohistochemical studies for alpha-smooth muscle actin should be made systematically in renal tissue biopsies. Myofibroblastic differentiation appears to play a significant role in the progression of renal failure and seems to be a useful marker of progressive disease. PMID- 10668217 TI - Apoptosis in cancer: therapeutic implications. AB - This review outlines the principal limitations of the mechanisms of active cell death (ACD, apoptosis) as the basis of tumorigenesis and the rationale of almost all therapies of malignancy. The concentration of cancer therapy in the direction of ACD induction is presented as both the result of progressive understanding of the mechanisms of apoptosis and that of the favourable tumor environment for ACD signal transmission. The latter property induces the by-stander killing of cancer cells, a fundamental mechanism because efficiency of all known methods of cancer treatment is far below 100%. Finally, recent results and hypotheses regarding cancer gene therapy based on final inductors of apoptosis and endogeneous ACD inhibitors in tumors are evaluated. PMID- 10668218 TI - A nuclear function for the tumor suppressor BRCA1. AB - The breast and ovarian cancer susceptibility gene BRCA1 has been recently cloned and revealed an open reading frame of 1863 amino acids, but a lack of significant homology to any known protein in the database has led to few clues about its functions. One of the first steps to investigate the function of BRCA1 was to define its subcellular localization. Several reports have led to contradictory findings that include: nuclear localization in normal cells and cytoplasmic in breast and ovarian cancer cells; nuclear in both normal and cancer cells; cytoplasmic and secreted to the extracellular space; present in tube-like invaginations of the nucleus; and colocalizing with the centrosome. As is apparent, the subcellular localization has been the most controversial aspect of BRCA1 biology and is a key point to uncover its functions. In this paper we review the published data on subcellular localization of BRCA1 with special emphasis on the antibodies and techniques used. We conclude that there is now overwhelming evidence to support a nuclear localization for BRCA1, both in normal and cancer cells. In addition, several BRCA1-interacting proteins have been isolated and they are preferentially located in the nucleus. Evidence supporting a physiological function for BRCA1 during DNA repair and transcriptional activation is also discussed. PMID- 10668219 TI - Genomic imprinting and Beckwith-Wiedemann syndrome. AB - Genomic imprinting is the parental-allele-specific expression of genes. Beckwith Wiedemann syndrome (BWS), a congenital overgrowth syndrome with increased risk of childhood tumors, is one of the well-known diseases caused by imprinted genes. The imprinted genes causing BWS are discussed in this review. PMID- 10668220 TI - Pathological changes of human unmyelinated nerve fibers: a review. AB - In the cutaneous nerves, unmyelinated nerve fibers outnumber the myelinated ones but are scarcely analyzed, especially at autopsy. This indifference toward the pathology of unmyelinated nerve fibers may be due to the necessity of electron microscopic analyses and, more importantly, the obscurity of pathological alteration of unmyelinated nerve fibers in aging as well as in peripheral nerve disorders. The aim of this article is to review (1) the normal appearance including postmortem changes, (2) the age-related changes, and (3) the pathological alteration in various neuropathic and non-neuropathic conditions, of unmyelinated nerve fibers in the sural nerve. For the complete analyses of sural nerve, qualitative and quantitative estimation of unmyelinated nerve fibers in all specimens should be recommended and it sometimes has an important diagnostic value. PMID- 10668222 TI - [IX Teaching Workshop on Family and Community Medicine. XIX National Congress of Family and Community Medicine. Abstracts]. PMID- 10668221 TI - Regulation of VEGF in the reproductive tract by sex-steroid hormones. AB - Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis. In adults, angiogenesis is an infrequent event in the normal tissue except in the female reproductive tract where angiogenesis occurs frequently during the cyclical repair and regeneration of the endometrium as well as in the ovary. Little is known about angiogenesis in the male reproductive tract. The role of VEGF in controlling reproductive tract physiology and the role of hormones in regulating this key regulator of angiogenesis is not well understood. Since reproductive tract physiology is largely under sex-steroid regulation, we have reviewed some recent studies describing the role of sex-steroid hormones in regulating VEGF. We have also included studies on the role of sex-steroids in regulating VEGF and angiogenesis in endometrial, breast and prostate pathologies. We have provided an extensive review of the classical VEGF and VEGF receptors with examples drawn from numerous studies in the literature using diverse biological systems to encourage similar studies in the area of reproductive tract physiology. It is speculated that such studies will provide insights into understanding the role of VEGF in reproductive tract development, causes of infertility, and cancer. Such knowledge would allow us to target VEGF for improving human reproductive tract abnormalities, for enhancing implantation and fertility, and for designing drugs for treatment of endocrine dependent cancers. PMID- 10668223 TI - [National Days of Laboratory Medicine of Russia, Moscow, October 12-14, 1999. Workshop: clinical laboratory at the brink of the XXI century: synthesis of traditions and novel trends (analysis, diagnosis, technology, and economics). Abstracts]. PMID- 10668224 TI - Detection of the viable myocardium. A perfusion scintigraphic study, before and after coronary bypass surgery in myocardial infarction patients. AB - OBJECTIVE: To compare single-photon-emission computed tomography (SPECT) imaging scans using 201Tl and 99mTc-MIBI in detection of viable myocardium, in regions compromised by infarction. METHODS: Thirty-two (59.3 +/- 9.8 years old and 87% male) myocardial infarction patients were studied. All had Q waves on the ECG and left ventricle ejection fraction of < 50%. They underwent coronary and left ventricle angiographies and SPECT before (including 201Tl reinjection) and after coronary artery bypass surgery (CABG). Improvement in perfusion observed after surgery was considered the gold standard for myocardial viability. RESULTS: Among 102 studied regions of the heart, there were 40 (39.2%) areas of transient perfusion defects in the conventional protocol with 201Tl and 52 (51.0%) after reinjection. Therefore, 12/62 (19.4%) more viable regions were identified by reinjection. Using 99mTc-MIBI, only 14 (13.7%) regions with transient defects were identified, all of which were seen also in 201Tl protocols. After surgery, 49 of a total of 93 regions analyzed (52.7%) were viable. Sensitivity, specificity, accuracy, positive and negative prediction values were, respectively, 201Tl SPECT scans--65.3%, 90.9%, 77.4%, 88.9% and 70.2%, reinjection protocol with 201Tl scans--81.5%, 81.8%, 81.7%, 83.3% and 80.0%, 99mTc-MIBI SPECT scans--20.4%, 90.9%, 53.8%, 71.4% and 50.6%. Logistic regression demonstrated that the reinjection protocol with 201Tl was the best predictor of viability (P < 0.001). CONCLUSION: Our data suggest the election of 201Tl for viability studies, especially when using the reinjection protocol. PMID- 10668225 TI - Brazilian multicenter study on efficacy and tolerability of trandolapril in mild to-moderate essential arterial hypertension. EMBATHE substudy with ambulatory blood pressure monitoring. AB - OBJECTIVE: A double-blind, placebo-controlled multicenter study involving 34 centers from different Brazilian regions was performed to evaluate the antihypertensive efficacy and tolerability of trandolapril, an angiotensin I converting enzyme inhibitor, in the treatment of mild-to-moderate systemic arterial hypertension. METHODS: Of 262 patients enrolled in this study, 127 were treated with trandolapril 2 mg/day for 8 consecutive weeks, and the remaining 135 patients received placebo for the same period of time. Reduction in blood pressure (BP) and the occurrence of adverse events during this period were evaluated in both groups. RESULTS: Significantly reductions in both systolic and diastolic pressures were observed in patients treated with trandolapril when compared with those on placebo. Antihypertensive efficacy was achieved in 57.5% of the patients on trandolapril and in 42% of these normal values of BP were obtained. The efficacy of trandolapril was similar in all centers, regardless of the area of the country. In a subset of 30 patients who underwent ABPM, responders showed a significant hypotensive effect to trandolapril throughout the 24 hour day. The adverse event profile was similar in both trandolapril and placebo groups. CONCLUSION: Our results demonstrate, for the first time in a large group of hypertensive patients from different regions in Brazil, good efficacy and tolerability of trando-lapril during treatment of mild-to-moderate essential systemic hypertension. PMID- 10668226 TI - Dynamic three-dimensional reconstruction of the heart by transesophageal echocardiography. AB - OBJECTIVE: To evaluate echocardiography accuracy in performing and obtaining images for dynamical three-dimensional (3D) reconstruction. METHODS: Three dimensional (3D) image reconstruction was obtained in 20 consecutive patients who underwent transesophageal echocardiography. A multiplanar 5 MHz transducer was used for 3D reconstruction. RESULTS: Twenty patients were studied consecutively. The following cardiac diseases were present: valvar prostheses--6 (2 mitral, 2 aortic and 2 mitral and aortic); mitral valve prolapse--3; mitral and aortic disease--2; aortic valve disease--5; congenital heart disease--3 (2 atrial septal defect--ASD- and 1 transposition of the great arteries-TGA); arteriovenous fistula--1. In 7 patients, color Doppler was also obtained and used for 3D flow reconstruction. Twenty five cardiac structures were acquired and 60 reconstructions generated (28 of mitral valves, 14 of aortic valves, 4 of mitral prostheses, 7 of aortic prostheses and 7 of the ASD). Fifty five of 60 (91.6%) reconstructions were considered of good quality by 2 independent observers. The 11 reconstructed mitral valves/prostheses and the 2 reconstructed ASDs provided more anatomical information than two dimensional echocardiography (2DE) alone. CONCLUSION: 3D echocardiography using a transesophageal transducer is a feasible technique, which improves detection of anatomical details of cardiac structures, particularly of the mitral valve and atrial septum. PMID- 10668227 TI - Risk factors for atherosclerosis in students of a private university in Sao Paulo Brazil. AB - OBJECTIVE: To characterize the risk profile for atherosclerosis (AS) in adolescents and young adults of a private university in Sao Paulo. METHODS: Clinical, nutritional, and laboratory parameters were evaluated in 209 students of both genders aged 17 to 25 years. In addition to determination of the lipid profile, the association of its abnormal values with other risk factors for AS was also investigated. RESULTS: Increased levels of total cholesterol, LDL-C and triglycerides (TG) were observed in 9.1%, 7.6% and 16.3% of the students, respectively, and decreased levels of HDL-C in 8.6% of them. Prevalence of the remaining risk factors analyzed was elevated: sedentary life style (78.9%); high intake of total fat (77.5%); high cholesterol intake (35.9%); smoking, hypertension (15.8%) and obesity (7.2%). There was an association between elevated LDL-C and TG levels and sedentary life style and body mass index. CONCLUSION: The high prevalence of risk factors for AS in young individuals draws attention to the need for adopting preventive plans. PMID- 10668228 TI - Comparison between right and left ventricular myocardia during the human fetal period. Stereological evaluation. AB - PURPOSE: To develop a stereological comparison between right (RV) and left ventricle (LV) myocardium during the third human gestational trimester. METHODS: Five human fetal hearts of the third trimester provided representative samples of 5 RV myocardium and 4 LV myocardium. The material was fixed in 10% buffered formaldehyde, and processed through routine methods. Fifteen microscopic fields were randomly chosen and counted in each ventricular myocardium using an "M-42" test system. The following stereological parameters were assessed: Vv (%), Lv (micron 2), Sv (micron 2/micron 3), Vp (micron 3), Nv (1/mm3) and total N. RESULTS: No significant difference between the stereological parameters of the myocardial structures assessed was evidenced, when comparing RV and LV. CONCLUSION: Right and left human ventricular myocardium are very similar during the fetal period at least in regard to their structural aspects. PMID- 10668229 TI - Surgical revascularization of posterior coronary arteries without cardiopulmonary bypass. AB - OBJECTIVE: To assess the results observed during the early postoperative period in patients who had the posterior coronary arteries revascularized without cardiopulmonary bypass (CPB), in regard to the following parameters: age, sex, bypass grafts types, morbidity and mortality. METHODS: From January 1995 to June 1998, 673 patients underwent myocardial revascularization (MR). Of this total, 607 (90.20%) MR procedures were performed without CPB. The posterior coronary arteries (PCA) were revascularized in 298 (44.27%) patients, 280 (93.95%) without CPB. The age of the patients ranged from 37 to 88 years (mean, 61 years). The male gender predominated, with 198 men (70.7%). The revascularization of the posterior coronary arteries had the following distribution: diagonalis artery (31 patients, 10%); marginal branches of the circumflex artery (243 patients, 78.7%); posterior ventricular artery (4 patients, 1.3%); and posterior descending artery (31 patients, 10%). RESULTS: Procedure-related complications without death occurred in 7 cases, giving a morbidity of 2.5%. There were 11 deaths in the early postoperative period (mortality of 3.9%). CONCLUSION: Similarly to the anterior coronary arteries, the posterior coronary arteries may benefit from myocardial revascularization without CPB. PMID- 10668230 TI - Rheumatic carditis treated with high doses of pulsetherapy methylprednisolone. Results in 70 children over 12 years. AB - PURPOSE: To report the result of patients treated with IV methylprednisolone divided into three groups and compare their follow-up during the last 12 years. METHODS: Seventy children with active rheumatic carditis (76 episodes) in heart failure Class III and IV (NYHA) were studied. The diagnosis was based on modified Jones' criteria. After ruling out infections and strongyloidiasis, treatment with IV methylprednisolone bolus was started three times a week until the laboratory tests became negative. Patients were divided into 3 groups, according to the time of hospital admittance: Groups 1, 2 and 3, comprising of 40, 18 and 12 children, respectively. RESULTS: Eighteen children in Group 1 (45%) were in their 1st attack: 2 series of pulsetherapy were used in 10 (25%), 3 in 9 (23%) and 4 in 21 (52%). In Group 2, 14 cases (77%) were in their 1st attack: 2 series were used in 7 (39%), 4 in 9 (50%) and 5 in 2 (11%). The echocardiogram showed a flail mitral valve in 12 (66%) of these patients (1 death occurred after mitral valvoplasty). In Group 3, 6 patients needed 5 or more series of pulsetherapy and a flail mitral valve was present in 5 (41%). One child underwent mitral valve replacement while still in the active phase, after 8 series of pulsetherapy, and another died. The number of patients who needed 5 or more series was significantly higher in Group 3. CONCLUSION: There were variations in the presentation and evolution of the cases during these 12 year. The established pulsetherapy protocol continues to be useful to treat severe cases. PMID- 10668231 TI - Evaluation of the heart rate and arrhythmias following the maze procedure for chronic atrial fibrillation. AB - PURPOSE: To assess the presence and the prevalence of arrhythmias and the variability of the heart rate in the medium-term postoperative period following the maze procedure for chronic atrial fibrillation (AF). METHODS: Seventeen patients with a mean age of 51.7 +/- 12.9 years, who previously underwent the maze procedure without cryoablation for chronic atrial fibrillation, were evaluated with the 24 hour electrocardiogram (ECG)--Holter monitoring from the 6th month after the operation. Valvular and coronary procedures were concomitantly performed. RESULTS: The mean heart rate during Holter monitoring was 82 +/- 8 bpm; the maximal heart rate was 126 +/- 23 bpm and the minimal heart rate 57 +/- 7 bpm. Sinus rhythm was found in 10 (59%) patients and atrial rhythm was found in 7 (41%). Supraventricular extrasystoles had a rate of 2.3 +/- 5.5% of the total number of heartbeats and occurred in 16 (94%) patients. Six (35%) patients showed nonsustained atrial tachycardia. Ventricular extrasystoles, with a rate of 0.8 +/- 0.5% of the total heartbeats, occurred in 14 (82%) patients. The chronotropic competence was normal in 9 (53%) patients and attenuated in 8 (47%). The atrioventricular conduction (AV) was unchanged in 13 (76%) patients and there were 4 (24%) cases of first degree atrioventricular block (AVB). CONCLUSION: After the maze procedure, the values for the mean heart rate, AV conduction and chronotropic competence approach the normal range, although some cases show attenuation of the chronotropic response, first degree AV block or benign arrhythmias. PMID- 10668232 TI - Cutting balloon angioplasty for intrastent restenosis treatment. AB - We describe here two patients with angiographic diagnosis of intrastent restenosis and regional myocardial ischemia. One stent restenosis was located in a native coronary artery and the other in a vein graft. Both were treated with cutting balloon angioplasty (CBA), inflated at low pressures. Angiographic success was obtained and both patients were discharged in the day after the procedure. Cutting balloon angioplasty using low inflation pressures achieved important luminal gains, in these two cases of intrastent restenosis. Further studies are necessary before the effectiveness of this procedure can be precisely defined. PMID- 10668233 TI - Myxoma of the mitral valve. AB - Only rarely do myxomas originate from the mitral valve. This is the report of a 49-year-old woman presenting with congestive heart failure. The diagnosis of an intracardiac tumor involving the anterior cuspid of the mitral valve was made by transesophageal echocardiography. The patient underwent surgery for tumor resection and plasty of the valve was made with reconstruction and preservation of the valve. The diagnosis of myxoma was confirmed by histology. This is the 23rd case of myxoma of the mitral valve reported in the literature. PMID- 10668234 TI - Systemic hypertension in heart transplant recipients. PMID- 10668235 TI - Beta-adrenergic pathway in healthy and hypertrophied hearts. PMID- 10668236 TI - [Organization of the network for the study of the Trypanosoma cruzi genome]. AB - Five years ago the Special Programme for Research and Training in Tropical Diseases (TDR) from the World Health Organization (WHO) launched the Parasite Genome Project. The aims were to obtain information on genome organization and gene discovery in five parasites, namely, Schistosoma, Filaria, Leishmania and Trypanosomas brucei and cruzi. Organization of research networks for each parasite under study, promotion of international collaboration and training of researchers in developing countries, were also main objectives of the programme. After five years, a large amount of information has been obtained, which is now available to researchers in the field. PMID- 10668237 TI - [Cruzipain, major cysteine proteinase of Trypanosoma cruzi: sequence and genomic organization of the codifying genes]. AB - Cruzipain is the major cysteine proteinase present in Trypanosoma cruzi, the parasite causing the American Trypanosomiasis, Chagas disease. The enzyme is encoded by a high number of genes (up to 130 in the Tul 2 stock) placed in head to-tail tandems, and located in two to four chromosomes. The simultaneous expression of several different genes results in the production of a complex mixture of isoforms. Those known as a group as "cruzipain 1" differ essentially at the level of the C-terminal domain, in their amino-acid sequence and in their type of N-glycosylation. The existence of a more distantly related cysteine proteinase, "cruzipain 2", has been demonstrated; it differs markedly, particularly at the level of the catalytic moiety. PMID- 10668238 TI - [Trypanosoma cruzi genome: transcriptional mapping and karyotype correlation; molecular characterization of a surface antigen from the Tc13 family]. AB - We describe herein the genome mapping of Trypanosoma cruzi, the etiological agent of Chagas' disease, by hybridization of a cosmid library gridded in high density filters with cDNA from an epimastigote expression library as probes. We also show the correlation of some cosmid contigs with parasite chromosomal bands. With libraries from the T. cruzi Genome Project we have characterized a new member of the Tc13 family belonging to the superfamily of trypomastigote surface antigens. Starting with a Tulahuen strain (Tul 2 stock) clone, homologous to these antigens, we have sequenced and characterized the complete gene in the CL Strain (CL Brener clone). We have also found homologies with different ESTs which characterization would lead to further knowledge of this antigen family. PMID- 10668239 TI - [Contribution of the Trypanosoma cruzi Genome Project to the understanding of the pathogenesis of Chagas disease]. AB - The Trypanosoma cruzi Genome Project (TcGP) is developed by a consortium of laboratories brought together by two international agencies, WHO/TDR and CYTED. The TcGP is an important tool for the study of Chagas disease. It provides researchers and clinicians with information about expressed parasite genes (see TcGP database, http:@gene.dbbm.fiocruz.br) and with an important number of genomic libraries and probes (ESTs). We show with four examples how the TcGP contributes to the understanding of the pathogenesis of this infection. 1) We demonstrate how to derive antigens from ESTs, an important feature regarding the generation of anti-cardiac receptor antibodies in chagasic patients. 2) We introduce concepts such as proteome vaccinome and pathonome that include all techniques needed to analyze quickly the growing amount of genomic information. 3) We show that the genomic regions that have been already sequenced give clues as to the plastic nature of the parasite nuclear genome. Finally, 4) we show how the genome sequences can be used to study the parasite DNA that is associated to the cardiac lesions. PMID- 10668240 TI - [Chagasic myocardiopathy: historical perspective]. AB - Considerable advances in the clinical pathological and pathogenic aspects of Chagas disease have been made since the Brazilian physician Carlos Chagas described the disease in 1909. The disease caused by the flagellate protozoon parasite Trypanosoma cruzi is transmitted to humans by a blood sucking triatomine and much less frequently by blood transfusion. It is estimated that 18 million are infected and that about 100 million people from Latin America are at risk of contracting T. cruzi infection. One of the most important contributions to the knowledge of Chagas' disease has been the recognition of the natural history of the disease, which can be divided into three well defined periods: 1. The acute stage; 2. An undetermined or undifferentiated stage and 3. The chronic stage. The primary infection (first stage) occurs mostly unrecognized and clinically apparent acute chagasic myocarditis may appear in less than 5% of the infected individuals, usually children living in endemic areas. The majority of the cases of acute myocarditis are mild and reversible. Autopsied cases of acute chagasic myocarditis are uncommon and correspond to exceptionally severe or fulminant forms showing diffuse myocardial damage with myocytolisis, degenerative changes of myocardial fibers and marked intersticial cellular infiltration. The acute clinical manifestations of the infected individuals include fever, muscular pain, sweating, swollen lymph nodes, hepatospienomegaly. Following this initial stage, all patients enter the undifferentiated or undetermined stage of the chronic period (second stage), which lasts between 10 to 20 years. Of these, 20 to 30% (depending on marked geographical differences) develop symptoms or signs of visceral damage conforming the cohort that enter the third stage. Although megaesophagous and megacolon are not uncommon (mainly in Brazil), the most frequent and important clinical manifestation is a dilated cardiomyopathy. Thus, 70% or more of the infected individuals will never show any clinical manifestation of the disease. The ajmaline test and the endomyocardial biopsy are, probably, the most sensitive methods to unmask latent forms of chagasic myocarditis during the undifferentiated stage. In the most advanced stages of chronic chagasic myocarditis, pathological findings are those of a dilated cardiomyopathy. At autopsy, the apical aneurysm with thrombus in it is a frequent and distinctive finding. The histopathological picture is that of an active and chronic microfocal and disseminated myocarditis. In some cases fibrosis may be confluent, which accounts for the electrocardiographic patterns of myocardial necrosis. The widespread distribution of cardiac lesions also constitute the substrate for atrioventricular and intraventricular conduction disturbances and for atrial and ventricular arrhythmias. The clinical diagnosis of Chagas' heart disease is based on a triad of: positive epidemiology, positive serology and a combination of clinical findings (suggestive electrocardiograhic abnormalities, apical aneurysm, cardiac enlargement). The electrocardiogram in the most advanced forms, usually shows sinus bradycardia, right bundle branch block with or without left anterior hemiblock, primary T wave abnormalities, pathological Q waves and multiform ventricular premature beats. The pathogenesis of the myocardial lesions of acute and also chronic chagasic myocarditis appears to be related in large part to autoimmune mechanisms. The lack of correlation between the location and number of parasitized fibers and the severity, type, and extension of degenerative and inflammatory lesions supports this assumption. Experimental and clinical studies have demonstrated the presence of antibodies directed against different components of T. cruzi and crossreacting with human antigens in patients with chronic chagasic myocarditis. Microvascular dysfunction, myocardial ischemia and autonomic nervous system impairment have also been implica PMID- 10668241 TI - Susceptibility and resistance to insecticides of Chagas disease vectors. AB - Chemical control of Chagas disease vectors appears to be the best practical way to reduce the incidence of the disease. DDT was initially tested in the 1950s for the campaigns of control of Chagas disease vectors. Its low level of effectiveness against triatomine caused the failure of these control actions. HCH was then introduced in the southern cone and Dieldrin in the north of Latinoamerica. Starting in the late 1960s anticholinesterasic organophosphorus and carbamate compounds were introduced in the control of Chagas vectors. The use of pyrethroid compounds began in 1980. This family of insecticides is now the most important tool in triatomines control because of its favorable toxicological properties. Other types of insecticides also studied for Chagas vector control were the insect growth regulators and the antifeeding compounds. Because of the mode of action of these insecticides they are now considered just a potential complement of neurotoxic insecticides for integrated programmes of Triatomines control. Innovative formulations such as fumigant canister and insecticidal paints have been successfully developed in Latinoamerica with the World Health Organization support. Resistance to insecticides of triatomines is not yet a great problem in Chagas vectors. However, some resistant strains to pyrethroids have been found in Rhodnius prolixus from Venezuela and in Triatoma infestans from Brazil. Some cases of T. infestans incipient resistance to deltamethrin have been detected in Argentina. According to the control tools now available it is possible to expect the interruption of vector transmission of Chagas disease in the near future. PMID- 10668242 TI - [Control campaigns against Triatoma infestans in a rural community of northwestern Argentina]. AB - Control campaigns against Triatoma infestans, the principal vector of Trypanosoma cruzi, have relied on the application of residual insecticides and have regarded the system as homogeneous. Reinfestation generally starts in peri-domestic residual foci or in preexisting foci that escaped spraying for diverse reasons. From these foci T. infestans adults actively invade other sites, or they are transported passively within objects or goods from infested communities. In the absence of additional spraying of insecticides after the attack phase, domiciliary reinfestation expanded exponentially to return to pre-spraying levels in 3-4 years in Amama, Santiago del Estero. However, the rate of recovery of T. infestans abundance was much lower than that predicted by a simple mathematical model or in experimental field studies. Reinfestation did not progress homogeneously within the village. Two longitudinal studies in Santiago del Estero revealed that the presence of T. infestans-infested peri-domestic sites increased the risk of domiciliary reinfestation. "Key" sites where reinfestation started early were goat or sheep corrals, pig corrals, chicken houses, and store-rooms. Peri-domestic reinfestation is likely the result of multiple factors, such as insecticide breakdown by climatic agents, and the greater surface and availability of refuge and hosts in peri-domestic rather than domiciliary sites. Such environmental heterogeneity generates insecticidal effects heterogeneity and increases the probability of persistence of T. infestans even under control pressures. Several studies showed that sustained surveillance after the attack phase decreased sharply the domiciliary colonization by T. infestans and the percentage of bugs infected with T. cruzi, and the local incidence of infected children. The sustained elimination of T. infestans demands a scientific approach that is not only centered in insecticide use but that also includes the environment and house-holders in their specific social and political scenario. PMID- 10668243 TI - [A tribute to Patricio Cossio. His contribution to the immunopathology of Chagas disease]. PMID- 10668244 TI - [Myocardial cell response to Trypanosoma cruzi infection]. AB - The aim of this study is to establish the response of cardiac myocytes to the infection with Trypanosoma cruzi. The role of myocardial cell proliferation on heart remodelation and the ability of these cells to produce nitric oxide and control intracellular parasite growth during T. cruzi infection were evaluated. The presence of proliferating cell nuclear antigen (PCNA) was determined in myocardial cells of Wistar rats infected with T. cruzi, resulting in a significant increase of PCNA+ labelling in all stages of disease. The ability of myocardial cells to control growth of intracellular parasites and the production of nitric oxide were evaluated in cultures of cardiac myocytes obtained from neonatal rats. Different combinations of cytokines were added to culture media. The number of cardiac cells displaying intracellular amastigotes was lower in cultures supplemented with IL-1b, TNF-a and IFN-g than with other cytokine combinations and controls. The addition of cytokines resulted also in an increase of nitric oxide production in both infected and non-infected controls. These results demonstrate that myocardial cells participate actively in the response of the heart to the infection with T. cruzi. PMID- 10668245 TI - [Pathogenesis of human chronic chagasic myocarditis]. AB - Studies carried out during the last decades provided evidence in support of an autoimmune pathogenesis for chronic chagasic myocarditis. This opinion was based on 1) the demonstration of molecular mimicry between parasite and host antigens, 2) the appearance of autoantibodies recognizing heart epitopes during the chronic phase of infection, 3) the induction of myocarditis and electrocardiographic alterations in animals immunized with whole parasites, parasite fragments or with biochemically-defined antigens, 4) the isolation from the heart of inflammatory infiltrates of B cells elaborating antibodies against myocardial antigens and 5) or of T cell clones reacting with heart epitopes and 6) induction of heart and nervous tissue alterations by transfer of lymphocytes from infected animals into naive syngeneic hosts. However, the characteristics of the inflammatory infiltrate in human myocarditis, displaying a wide variety of cells, many of them not involved in autoreactivity, such as the presence of giant cell granulomas and abundant eosinophils, as well as its focality and asynchrony, and the frequent association with pericarditis, casts doubts about the possibility of autoimmunity being responsible for the perpetuation of the myocarditis. This is supported by the recent observation that treatment of asymptomatic patients with trypanocidal drugs prevents the development of cardiopathy and that parasite components, either antigens or genomic fragments, are present at the site of the inflammatory lesions. On the basis of this new evidence, other alternative pathogenetic mechanisms should be sought to explain the appearance of a polymorphic long lasting myocarditis that needs the presence of tiny fragments of parasites to develop. In addition to the well known immunological pathogenesis, the link between such a small amount of parasite components, below the level of microscopic detection, and the induction of such an extensive inflammatory infiltrate, represents interesting avenues for research in the near future. PMID- 10668246 TI - [Trypanosoma cruzi pathology. Strain dependent?]. AB - There is agreement today about the role that the characteristics of the population of Trypanosoma cruzi play in the pathogenesis of the different clinical forms of Chagas disease. In our laboratory, we have studied the outcome of the infection of mice with two populations with polar biological behaviour: RA and CA-I. We have demonstrated that the neuromuscular damage is, in part, mediated by different T cell subsets. We have also observed that the T cell phenotype responsible for the pathology and the targetted tissues depend on the parasite population. Although we found no differences regarding the reactivity of IgG to native nerve structures in sera from mice infected with either strain, it is presumed that the humoral response would play an additional role in the development of strain-dependent neuromuscular pathology since passive transfer of sera from mice infected with RA triggered alterations of the nerve action potential whereas sera from CA-I-infected mice did not. We have also detected a reduction in the fertility of female mice infected with CA-I/K98, whereas females infected with RA showed no difference in comparison with uninfected controls. However, congenital transmission was only observed in mice infected with RA. The differences observed in fertility, in newborn survival, and in the number of fetal resorptions in mice infected with the myotropic strain could be attributed to the uterine inflammatory response, since no estrous alterations were observed between infected and control groups. PMID- 10668247 TI - Overview of molecular mechanisms in chagasic cardioneuromyopathy and achalasia. AB - Evidence accumulated by our investigations over the years give adequate proof for the existence of circulating antibodies in Chagas disease which bind to beta adrenergic and muscarinic cholinergic receptor of myocardium. The interaction of agonist-like antibodies with neurotransmitter receptors, triggers in the cells intracellular signal transductions that alter the physiological behaviour of the target organs. These events convert the normal cells into pathologically active cells. The interaction of antibodies with heart beta adrenergic and cholinergic receptors triggers physiologic, morphologic, enzymatic and molecular alterations, leading to tissue damage. The analysis of the prevalence and distribution of these antibodies reveals a strong association with cardiac and esophageal autonomic dysfunction in seropositive patients in comparison with those without alteration of the heart and esophagus autonomic disorders: therefore, the presence of these antibodies may partially explain the cardiomyoneurophathy and achalasia of Chagas disease, in which the sympathetic and parasympathetic systems are affected. The deposit of autoantibodies behaving like an agonist on neurotransmitter receptors, induceds desensitization and/or down regulation of the receptors. This in turn, could lead to a progressive blockade of neurotransmitter receptors, with sympathetic and parasympathetic dennervation, a phenomenon that has been described during the course of Chagas cardioneuropathy and achalasia. The clinical relevance of these findings is the demonstration, using biomolecules, of a strong association between the existence of circulating autoantibodies against peptides corresponding to the second extracellular loop of the human heart beta, adrenoceptor and M2 cholinoceptor in chagasic patients, and the presence of dysautonomic symptoms, making these autoantibodies a proper early marker of heart and digestive autonomic dysfunction. PMID- 10668248 TI - [TH1 response in the experimental infection with Trypanosoma cruzi]. AB - Specific antibodies and the activation of phagocytic cells by IFN-gamma are the key elements of the immune response involved in protection of the T. cruzi infected host. The central role of the IFN-gamma in vivo seems to be the activation of the inducible nitric oxide synthetase of macrophages (iNOS) and the production of nitric oxide (NO degree) for the intracellular destruction of the parasite. Interleukin 12 (IL-12), the cytokine that stimulates NK cells for IFN gamma production, seems to trigger the TH1 response in the acute phase. Other cell types, such as lymphocytes Thy-1+CD4-CD8-, CD4+ and CD8+, are also involved in IFN-gamma production. The down regulation of the TH1 response could in part depend on the decrease in the macrophage activation, as a result of the controlled parasite burden, and on the production of IL-10 and transforming growth factor beta (TGF-beta). The protective TH1 immune response seems to be also related to both the tissue damage and the alterations of the immune response observed during the infection. We studied the kinetics of both NK cell activity, and the production of IL-12 and/IFN-gamma by spleen cells, as well as the seric levels of these cytokines, in BALB/c and C3H mice infected with T. cruzi, Tulahuen strain. In the spleen, we found that the production of IL-12 and the NK cell activity increased in the very early acute infection, and that in C3H the effect was higher than in BALB/c mice. IFN-gamma increased in C3H at the same time, but in the BALB/c strain it increased later in the acute phase. The infection induced a very early increase in the seric levels of IL-12, that remained high throughout the acute phase, in both mouse strains. However, the levels of IFN-gamma in the serum increased a few days before the peak of parasitemia, reaching higher values, and earlier, in BALB/c than in C3H mice. Surprisingly, in the chronic infection IL-12 production remained high in both mouse strains, but IFN-gamma production was only observed in BALB/c mice. The immune response was predominantly TH1 in both mouse strains, in spite of the higher susceptibility of BALB/c compared to C3H. The early control of the parasite burden could be evaluated as the expression of the TH1 response in spleen cells, while the seric levels of IL-12 and IFN-gamma would be related to the induction of tissue damage. Our data indicate that the protective TH1 immune response has a different expression according to the host-parasite relationship, and that the factors controlling the response are of primary importance to determine the quali- and quantitative expression of IL-12/NK/IFN-gamma as well as their involvement in resistance and tissue damage. PMID- 10668249 TI - [Control of the transmission of Trypanosoma cruzi in Argentina 1999]. AB - Approximately 2 million people in Argentina are infected with Trypanosoma cruzi, the etiologic agent of Chagas disease, thereby constituting the major tropical disease in the country. As in other six Southern Cone countries, Triatoma infestans is the only or major vector of T. cruzi among human and domestic animals. In Argentina, a vertically structured National Chagas Control Program was established in 1962. Such a program pursued the elimination of domestic and peri-domestic populations of T. infestans through insecticidal spraying, and the serological control of blood donors to prevent transfusion-related infections. This program strongly reduced the nation-wide serological prevalence of T. cruzi in the population. For example, in 18 or 20 year-old men drafted into military service, the seroprevalence decreased from 10.1% in 1964 for those who had been born in 1944 to 1.9% in 1993 for those born in 1975. However, the vertical strategy failed to reach and sustain the surveillance phase in widespread rural areas with disperse populations due to its intrinsic limitations and the reduced priority level assigned to rural health programs. An alternative, horizontally structured control strategy of T. infestans was developed and assayed in the Province of Santiago del Estero between 1985-1989, and 1991-1992. The projects demonstrated that insecticidal spraying carried out with community participation combined effectiveness and commitment in such a way as to produce a strong impact on house reinfestation and the extension of the area under entomological surveillance. This experience has been transferred in a chain of responsibilities to the personnel of the National Chagas Control Program, using participating workshops, procedural guidelines, and practical training. This personnel transferred the strategy using similar methods to the field health care agents and volunteers chosen by their own communities (community leaders). After the workshops, the leaders received all the materials needed to install and develop the ongoing surveillance activities: third generation pyrethroid insecticides, manuals, hand-operated sprayers, and sensor boxes to detect domiciliary infestations. From 1993 to 1998, a total of 15,000 health care agents or community leaders were trained. A total of 675,000 houses were sprayed with residual insecticides in the attack phase, and 850,000 houses entered the surveillance phase. This is the first time that such large coverage has been accomplished in Argentina. The network of laboratories installed a quality assurance program to current serological procedures applied to blood donors, organ transplant, and the detection and treatment of newborns to women sero reactive for T. cruzi in Argentina. We expect to interrupt the vector-mediated transmission of T. cruzi in the next 18 months, but the sustainability of such a program depends on, at least, additional work with the community to achieve a change of attitudes and practices related to house infestation for the next 10 years. A social effort will be needed to cover those expenses, but the expected economic returns exceed largely the cost of any such program, as suggested by cost-benefit studies. To illustrate, the annual treatment costs of one Chagas patient can help maintain 25 households free from triatomine bugs in Argentina. PMID- 10668250 TI - [Elimination of vectorial transmission of Chagas disease in Brazil]. AB - The control of vectorial transmission of Chagas disease in Brazil has been systematized and structured into a national program since 1975 when regulations were set up on the basis of entomological and sero-epidemiological studies which permitted the delimitation of areas at risk of vectorial transmission in the whole country as well as the orientation of the chemical control of domiciliary vector populations. The authors present the original data collected throughout the years comparing them with the present data. The evaluation reveals a virtual interruption of the transmission for Triatoma infestans and the remaining possibility of transmission, at very low levels, for native vector species in different areas of the country. It is emphasized that it is important to maintain constant entomological vigilance in order to prevent the reestablishment of transmission. PMID- 10668251 TI - [Current status of the control of Chagas disease in Colombia]. PMID- 10668252 TI - [Towards the elimination of the transmission of Trypanosoma cruzi in Honduras and Central American countries]. AB - Central America is composed of seven countries: Belice, Costa Rica, El Salvador, Guatemala, Honduras, Nicaragua and Panama. Chagas disease exists in all seven countries, but with major prevalence in El Salvador, Guatemala, Honduras and Nicaragua. The main species of triatomine vectors are: Rhodnius prolixus, Triatoma dimidiata y Rhodnius pallescens. In 1997 the Central American countries launched an Initiative for the Vectorial and Transfusion Transmission Control of Chagas disease. The objectives of the Initiative are: 1. elimination of Rhodnius prolixus, 2. control of Triatoma dimidiata and 3. serological screening for Trypanosoma cruzi of 100% of the blood donors. This Initiative is supported by the Resolution for "The Elimination of Transmission of Chagas Disease", of the World Health Assembly in 1998. PMID- 10668253 TI - [Progress towards the interruption of transmission of Chagas disease in the southern countries]. AB - The epidemiological and entomological data and the trends observed in the decreasing of the incidence of infection in young age groups indicate that only ninety years after the discovery of Chagas disease, the control of vectorial and transfusional transmission has reduced the incidence by 70% in the Southern Cone countries (Argentina, Bolivia, Brazil, Chile, Paraguay and Uruguay). This has been accomplished thanks to the political and financial engagement of the concerned governments who have invested US$340 millions since 1991 to the present. The initiatives to interrupt transmission of Chagas in the Andean countries and the Central American countries have begun their activities in 1997 and the evolution of the control operations allows to forecast the complete interruption in these areas before the year 2010 to comply with the mandate of Resolution WHA. 52.14 of the World Health Assembly in May 1998. PMID- 10668254 TI - [Risk of Chagas disease through transfusions in the Americans]. AB - The safety of blood transfusion depends on a country's laws, decrees and/or regulations concerning the collection, production and use of blood and blood derivatives. It also needs governmental enforcement of those instruments, as well as trained health professionals to obtain blood and produce blood derivatives, following total quality control procedures both at collection and production, and use. By 1998, all Latin American countries had laws, decrees and/or regulations that governed the production and use of blood, with the exception of El Salvador and Nicaragua. During the past six decades, economic need in Latin America has promoted migration to urban areas. Consequently, at present time, more than 60% of the population live in cities, which increases the probability of finding blood infected by Trypanosoma cruzi among donors. Unless all the blood from infected donors is discarded, the possibility of transmitting infection by transfusion remains. Moreover, infection by T. cruzi through transfusion is a potential problem in developed countries, now that tens of thousands of individuals from Latin America have migrated to the United States, Canada, western Europe, Australia and Japan. When donors are not screened for T. cruzi, the risk of transfusing infected blood is greater at higher prevalence rates of infection in the donor population; it also increases with the number of transfusions received by the recipient. In 1993, Bolivia presented the highest risk of receiving infected blood and becoming infected with T. cruzi; this country was followed by Colombia, El Salvador and Paraguay. As the coverage of HIV screening became almost universal, the probability of receiving blood infected by HIV and becoming infected was low in all countries. In the case of hepatitis B (HVB), the highest probability of infection was in Bolivia, Nicaragua and Guatemala. This probability was even greater for Hepatitis C (HVC), given the low coverage of donor screening in all countries. In absolute numbers, the highest potential for occurrence of cases of T. cruzi infection were present in Bolivia, the greatest number of HVC cases in Colombia, and the most cases of HVB in Nicaragua. Only in two countries, Bolivia and Colombia, HIV could be potentially transmitted by blood transfusion. Although the situation has improved since 1993, and 100% of donors are being screened for T. cruzi in Argentina, Colombia, Ecuador, El Salvador, Honduras, Paraguay, Uruguay and Venezuela, success will only be assured by: total enforcement of the law by governments; implementation of altruistic and volunteer blood donations, exclusively; 100% of donors are screened for communicable diseases; the collection, processing and use of blood strictly follow quality control norms; reagents used in diagnosis are adequate, and the use of blood and blood derivatives is limited to cases where it is only absolutely necessary. PMID- 10668255 TI - [Transfusion transmission of Chagas disease in Honduras and other Central American countries]. AB - The transmission of Trypanosoma cruzi through blood transfusion is the second most important way of acquiring Chagas disease. This form of transmission transcends the vectorial transmission in many geographical areas. Honduras has developed a successful program for the control of transfusional transmission, based on the serological screening of blood donors, which is supported by law, making it mandatory. This experience has been extended to other Central American countries, supported by PAHO. Actually in the framework of the Initiative of the Central American countries for the elimination of the transmission of Chagas disease, launched in 1997, the control of transfusional transmission is becoming a reality. PMID- 10668256 TI - [Control of congenital transmission of Trypanosoma cruzi in Argentina]. AB - The vertical transmission of Trypanosoma cruzi has been augmenting its relative importance as vector and transfusion-mediated transmission routes have been, and continue to be, increasingly controlled. The vertical transmission of T. cruzi cannot be prevented; but early detection and treatment of congenital infection achieve cure rates close to 100%. In Argentina, the Subprogram of Control of Pregnant Women examined 58,196 women from 13 provinces in 1997 and found a 9% seropositivity to T. cruzi. In spite of such high maternal prevalence rates of T. cruzi, only a small proportion of live newborns to infected mothers acquires the infection. The probability of vertical transmission was 1.9% (range: 0.1% to 3.5%) in surveys carried out in the '70s, and 2.5% (range: 0.7% to 10.4%) in others conducted in the '70-'80s. Other more recent studies in Argentina estimated the probability of transmission in 2.6%-6.7%, but studies from Paraguay estimated 10.5% by PCR or serodiagnosis. The microhematocrit technique is the recommended parasitologic method to detect congenital infection. Routine serodiagnosis that detects IgG against T. cruzi is only helpful after the newborn reaches 6 months of age. Detection of specific IgM using recombinant antigens and PCR constitute excellent alternatives, but their feasibility from operational and cost-effective viewpoints in affected endemic areas remains to be considered. In a longitudinal project carried out in Maternidad Nuestra Senora de la Merced in the city of Tucuman between 1992-1994, the majority of congenital cases were asymptomatic. They were diagnosed through the microhematocrit technique, but a number of cases could only be detected later as a result of the parasitological and/or serological follow-up. Of a total of 32 newborns infected with T. cruzi who were treated with nifurtimox or benznidazole, 30 had a negative microhematocrit and serodiagnosis between 6 months and 2 years post-treatment. The magnitude of congenital transmission, and its associated morbidity and mortalidad, largely justify the efforts needed to detect T. cruzi infection in the mothers and newborns. This project demonstrated that the transmission of T. cruzi can be successfully controlled at a provincial scale through a specific program inserted in the primary health care system or at the first level of attention. The congenital transmission of T. cruzi clearly represents a public health problem in areas that in the past were of active transmission, even years after being under entomologic surveillance. PMID- 10668257 TI - [The transmission de Chagas disease in Salta and the detection of congenital cases]. AB - Data on the prevalence of Trypanosoma cruzi infection is presented for the province of Salta, Argentina. Special emphasis is given to the detection of congenital transmission and to the economic benefits of preventing Chagas' disease. Seroepidemiological data obtained from 20 year old army draftees revealed a reduction, from 22.7 to 11.11% between 1964 and 1985. In university students, a rate of 0.96% was found in 1998. Surveys carried out during 1996 showed that more than 15% of the pregnant women analyzed carried T. cruzi infection, particularly in the north of the province. This situation brings about a high risk of appearance of congenital cases and represents an opportunity to test the most adequate strategies for detection. By applying systematically microhematocrit, hemoculture and PCR methods, to umbilical chord blood, an increase in the early detection of congenitally infected babies is being achieved. In 1992-94, very high seroprevalence rates of infection were found among indians of the Chaco region of Salta. The overall rate was 37%, but there were 5 localities where more than 54% of the population was infected. These numbers indicate that, in vast areas of the provincial territory, fight against vector bugs must not merely consist of surveilance activities, but rather of renewed spraying attacks. The fight must include control of pregnant women and blood banks. An economic analysis of the economic return, calculated only for spraying activities and for the Department of Anta (Salta), indicated a net present value of over 7 million dollars and an internal rate of return exceeding 60%. PMID- 10668258 TI - [The chemotherapy of Chagas disease]. AB - To date, Chagas disease has defied all attempts to develop an efficient and safe chemotherapy. Drugs effective on T. cruzi as trypanocidal agents may be classified as (a) drugs of extensive clinical use: Nifurtimox and Benznidazole; (b) drugs of restricted clinical use: azoles (e.g. Ketoconazole, Econazole; Miconazole); Amphotericin B; Allopurinol, Allopurinol ribosides and Primaquine; (d) drugs effective on T. cruzi and in experimental Chagas disease (murine model): alkyllysophospholipids; 5-amino-imidazole-4-carboxamides; bisbenzyl isoquinolines; cruzipain (crucein) inhibitors; Gossipol; phenothiazines; d) drugs effective in vitro without other reported effects, acridines, actinomycin D, Crystal Violet (gentian violet), diterpenes (Mikania obtusata); N,N'-dimethyl-2 propen-1-amine, epoxidienthiol carbamates, Fe-chelators, guanyl hydrazones, o naphthoquinones (beta-lapachone); quinoids (miconidine; tingenone); Olivacine, phenazine methosulfate, phenoxi-phenoxyl drugs, Proadifen, pyridinium azolate betaines, sesquiterpenes (Lychophora sp), sesquiterpene lactones, tetrahydrocarbazoles, DL-alpha-trifluoromethylarginine, triphenylmetane dyes. It is generally agreed that Nifurtimox and Benznidazole (a) are effective on acute Chagas' disease, but may not be effective in the chronic phase; (b) their effect depends on the susceptibility of T. cruzi strains to the drug; (c) they produce adverse effects in patients that may prevent prolonged treatments; they are genotoxic and produce biochemical damage in the mammalian tissues. Redox-cycling of Nifurtimox and Benznidazolee generates "reactive oxygen species" which explain the biological effects. At variance with the mammalian host, T. cruzi is deficient in antioxidant enzymes which are essential to prevent oxidative damage. Azoles are effective inhibitors of T. cruzi growth in vitro and in vivo since they inhibit sterol C14-delta 24(25) demethylase, an enzyme catalysing ergosterol production. Azoles reduce parasitemia and extend the survival of infected mice but do not produce parasitological cure and their clinical effectiveness is questionable. Allopurinol allopurinol ribosides and related compounds inhibit T. cruzi hypoxantine-guanine ribosyl transferase, thus preventing the synthesis of adenylic and guanylic acids and also DNA. They reduce parasitemia and negativize xenodiagnosis but these effects may not be permanent, which invalidates their clinical use. Cysteine-protease inhibitors recognize T. cruzi protease (cruzipain, crucein) active site, thus allowing a covalent linkage with the inhibitor. These peptide inhibitors are effective in acute and chronic murine models. Phenothiazines inhibit trypanothione reductase and a specially favoured fit is a small 2-substitued 2-chloro and 2-trifluoromethyl with a remote hydrophobic patch. The essential phenotiazine nucleus can adopt more than one inhibitory orientation in its binding site. Phenothiazines are promising trypanocidal agents for the treatment of Chagas' disease. The methodology for developing new drugs for the treatment of Chagas' disease is discussed. PMID- 10668259 TI - [Treatment of Trypanosoma cruzi infection in the indeterminate phase: experience and current guidelines in Argentina]. AB - An effective treatment for Trypanosoma cruzi infection has been investigated, since the 30s. The goals of the specific treatment against T. cruzi infection are, at the individual level, to eliminate the parasite, and to reduce the probability of developing Chagas disease. At the end of the 60s and at the beginning of the 70s, two compounds were clinically investigated in Argentina: Nifurtimox and Benznidazole. After the approval by the Ministry of Health, in 1983 the first guidelines for the treatment of T. cruzi infection were proposed and approved. These guidelines recommended the treatment of cases in the acute phase. Due to the publication of new information in support of the utility of these drugs for treating cases in the indeterminate phase of Chagas disease, in 1997 the original guidelines were revised and new procedures were approved. At present, the treatment is recommended for: 1) all patients undergoing the acute phase; 2) children and young people undergoing the indeterminate phase; 3) adult patients undergoing the indeterminate phase or with incipient heart lesions; 4) laboratory accidents and during surgery, and 5) organ transplant recipients or donors. The general clinical laboratory control is needed for the intra-treatment monitoring of the patient. Titration of specific antibodies with monospecific antigens has been shown to be an adequate marker of therapeutic efficacy. PMID- 10668260 TI - [Trypanocidal effect of cysteine protease inhibitors in vitro and in vivo in experimental Chagas disease]. AB - Endemic in most American countries, Chagas' disease causes high morbidity and mortality. Recent experimental and clinical evidence shows the importance of chemotherapy in both the acute and chronic phases of this disease. However, treatment is yet limited by the toxicity associated to available drugs. This review describes the design, evolution, and selection of dipeptides that interrupt the intracellular cycle of T. cruzi and cure acute experimental infections in laboratory animals. Peptido-mimetic inhibitors specifically bind cruzain, a T. cruzi cystein protease. The inhibitors cause alterations in the Golgi complex and ER, accumulation of unprocessed enzyme within Golgi cisternae, and decrease of mature cruzain within lysosomes. The most effective compound, N Pip-F-hF-VS phi, cured an acute lethal infection in experimental animals. Myocardial lesions, lymphocyte infiltration and intracellular amastigote clusers were absent in treated animals. Preliminary toxicology and pharmacokinetic analyses suggest the lack of toxicity associated to high doses and prolonged treatment regimes. Protease inhibitors may soon become good chemotherapeutic alternatives for acute and chronic Chagas' disease. PMID- 10668261 TI - Polymerase chain reaction (PCR) as a laboratory tool for the evaluation of the parasitological cure in Chagas disease after specific treatment. AB - The evaluation of the treatment for chronic Chagas disease faces the absence of any clear-cut criterion of cure. The low degree of parasitemia and the persistence of positive immunologic reactions represent some of the difficulties involved in addressing therapeutic efficacy. Our aim was to define whether PCR could be used as a laboratory method for evaluating cure in Chagas disease after specific treatment. We tested the utility of PCR amplification of the variable regions of minicircles from Trypanosoma cruzi kinetoplast DNA, in 76 xeno positive chronic Brazilian patients who have been treated with benznidazole in Mambai (Goias State) and Sao Felipe (Bahia State). We observed a positive amplification result in only 25 out of 76 treated patients (33%). Therefore, the performance of one single PCR after therapy revealed parasite clearance in 67% of the treated individuals, while xenodiagnosis was negative in 84%. These observations suggest that PCR is the most sensitive technique available for direct detection of T. cruzi in chagasic patients and that it can be a very useful instrument for the follow-up of patients after specific chemotherapy. In this sense, we are now developing a quantitative approach based on the use of fluorogenic probes and real-time measurement of the amplification reaction (TaqMan technology) in order to precisely estimate the parasite load in chronic chagasic patients before and after treatment. This may be the basis for the future establishment of reliable criteria of cure for patients undergoing therapy. PMID- 10668262 TI - Future prospects for the chemotherapy of Chagas' disease. AB - Over the last two decades, progress towards new drugs for the treatment of Chagas' disease has been disappointing. However, as a result of the parasite genome sequencing projects, the possibility of identifying novel drug targets through genomics, proteomics and bioinformatics has never been better. Progress towards the development of novel therapeutics, from target identification and validation by chemical and genetic means through to rational drug design, is illustrated with reference to the metabolism and functions of trypanothione, with particular emphasis on trypanothione reductase, one current drug target of choice. PMID- 10668263 TI - [Primary glomerulopathies]. PMID- 10668264 TI - [Anatomy of the splenic artery]. AB - Due to the importance and permanent improvements as regards the treatment of different diseases involving abdominal organs such as the abdomen, pancreas and spleen. I was indined to investigate, both from the anatomic and surgical point of view, one of the collateral arterial branches of the celiac trunk, which is possibly considered to be one the least studied and, historically one of the most forgotten by the vessels irrigating the liver and stomach. Considering the fact that anatomic understanding is the starting point of medical knowledge, and that its conquests give it permanently new fundaments, I was determined to do research, intensely and thoroughly on splenic artery. This research work relates in detail, on the corresponding chapters, the material and methods used, which consist on human bodies and foetus fixed with formol and in a fresh condition. These human bodies and foetus were submitted to dissection techniques of inter arterial injection with resin, and then they were corroded with acid and angiographic studies. Finally selective arteriographic tests were performed on living beings by means of digitalis removal of the splenic artery. The following chapter deals with the results obtained from saied research work, showing that the splenic artery originates, in all cases, from the celiac trunk, and this artery is the most important with an average length of 10.6 centimeters and of 2.3 flexuosities in all. This chapter also enumerates the relationships existing between the splenic artery and neighboring organs, the homonymous vein, its collateral and terminal branches which in the hundred percent of cases showed one superior and one inferior, to immediately subdivide in different ones at the level of the splenic pedide. The collateral branches found were the following: arterial pancreatic branches found in the 73% of the cases with an average of 1.8 arteries in all. posterior esophaguscardiotuberosity artery appearing in a 33% of the cases. superior polar artery appearing in a 53% of the cases. inferior polar artery appearing in a 33% of the cases. the short vessels appeared in the 100% of the cases, from which in a 73% were superior short vessels and in the remaining 27% inferior short vessels. the left gastroepiploon artery appeared in all the cases considered. I have discussed the anatomic importance of the splenic artery and gland with their multiple variations and relationships. I have also considered the splenic pedicle, depending on the rear fixation which has the extremity of the pancreas and the splenic hillum of 2.2 centimeters, with final figures ranging from 0 to 4 centimeters. I have also mentioned the appearance of a 10% of super numerary spleens. Finally in our experience with splenic segmentation and with corrosion pharmaceutical preparation and arteriographic studies as the basic ingredients, we have found that in a 62.5% they have two segments, in a 17% they have three segments, in a 12.5% they have four segments, in a 12.5% they have four segments and in an 8% they have five segments in all, in these cases generally due to the high importance of the polar arteries. Likewise, it is a truth universally accepted that splenic circulation is terminal and that spleen division into segments is separated by non-vascular levels. PMID- 10668265 TI - [Comparative structural study of the glial blastomas of the central nervous system]. AB - The objective of our work is to compare 2 diagnostics methods and demonstrate the similarities of the different Central Nervous System Glial Tumor Classifications. Immuno-marking techniques for gliofibrilar acid protein (GFAP), and special silver impregnations to compare the results according to the last histogenetic interpretations were used. 95 Gliomas were studied with hematoxiline-eosine and silver techniques, and in 58 randomized selected cases the technique for GFAP was also used. Both methods showed the same results in the diagnosis of Glial Tumors whose cells possess gliofibrils: Glioepitelioma (Ependymoma), Glioblastoma, Astroblastoma, and Astrocytoma. In the Oligodendrioglomas whose cells possess microtubules but not gliofibrils, the silver techniques marked these cells and their prolongations while the technique for GFAP did not. Regarding Gliomas, there are similarities by comparing the Del Rio Ortega-Polak classification with those of the WHO and other authors. There are differences only in the "names" of some tumors or in their histogenetic interpretation, which are not substantial and are confined to the "Polar Espongioblastoma" and "Gliosarcoma". Therefore, we think that the recognition of the Del Rio Hortega-Polak classification is justified. PMID- 10668266 TI - [Human papillomavirus detection in oral cancer lesions in the city of Cordoba]. AB - Oral cancer is a process that involves different etiological factors and mechanisms in the light of current view of viral cocarcinogenesis. Evidence from histology and DNA hybridization studies suggests that HPV is engaged in oral carcinogenesis. The Pathology Laboratory of the Dentistry School, National University of Cordoba, admits approximately 20% of all patients with cancerous lesions in this city. In the January 1992-December 1997 lapse, we examined 1950 biopsies with oral lesions, 4.77% (93/1950) of which were malignant neoplasms, 79.57% (74/93) were oral carcinomas. Thirty-three oral carcinomas (44.6%; 33/74) were selected at random and included in this study, 33 cells smears of normal oral mucosa of controls individuals were included. They were analyzed by conventional light microscopy and an in situ hybridization technique for the detection of HPV. Data were analyzed with chi square test. The prevalence of HPV among the 33 cancer samples studied was 27.27%, 9/33 tested positive for HPV in low stringent conditions. Only one was positive in high stringent condition for HPV16, a verrugous carcinoma. No HPV-DNA was detected in cells smears of controls. Among the HPV positive, 3/9 (33.33%) were squamous carcinomas and 5/9 (55.56%) were verrugous carcinomas. Only one was a melanoma. Verrugous carcinoma was the carcinoma most associated with the HPV infection (x2 = 20.5; 95% level of confidence). This would indicate a major role of HPV in the pathogenesis of verrucous carcinomas. The viral prevalence found in cancerous lesions reinforces the concept of heterogenic natures of oral cancer. HPV is a circumstance that increase the probability of malignancy, and when reducing, diminish the frequency of cancer. PMID- 10668267 TI - [Importance of the benzylpenicillin nucleus and the side chain of the beta lactams. Demonstration by skin tests and RAST in penicillin allergic patients]. AB - We studied 30 patients with beta-Lactams allergy demonstrated by clinical findings. The aim of this work was to determine the capacity of the beta-Lactams nucleus and the side chain in the induction of specific IgE to BPO, Ax, Amp, performed by intradermal skin test and RAST. The patients were divided by clinical manifestations in: 1-Accelerated reactions (n:19); and 2-Immediate reactions (n:11). The Prick tests were performed with BPO-PL, Ax-PL, Amp-PL, MDM BP, MDM-Ax, MDM-Amp. The accelerated group presented BPO-PL (+) in 2 cases, Ax-PL & Amp-PL (+) in 4 cases, and all of the reactives were (+) in 13 out of 19 cases. The immediate group presented MDM-BP (+) in 10 out of 11 cases and MDM-Amp was (+) in 1 out of 11 cases. The RAST's were performed in all patients(n:30). In accelerated group were (+) to BPO-PL in 13 out of 19 cases, to Ax-PL in 3 out of 19 cases, to Amp-PL in 1 out 19 cases, to BPO-PL and Ax-PL on overlap in 1 out of 19 cases, and 1 case was negative to all reactives. The immediate group presented RAST's negatives in 11 out of 11 cases. The control group(n:20) presented Prick (+) to Ax-PL in 1 out of 20 cases, and the others reactives were negatives in all cases. The RAST's to all reactives were (-) in 20 out of 20 subjects. These results indicate that BPO was the most important determinant, and the side chain of the Ax or Amp were others determinants of the beta-Lactams drugs. These determinants induced specific IgE, and in rare occasions appears specific IgE for two different determinants on overlap in the same patient. The intradermal skin testing is the method of choice to study the penicillin allergies, because non satisfactory RAST's have yet been developed for minor determinant-specific IgE antibodies. PMID- 10668268 TI - [Isolation of Vibrio cholerae non 01 from patients with acute gastroenteritis]. AB - It was assay by biochemical and immunological tests, strains of V. Cholerae non 01 non-0139 isolated from two patients of the province of Cordoba, Argentina. They showed episodes of acute gastroenteritis. Strains non-01 non-0139 were isolated from water samples ingested by patients. We conclude that strains identified from patients would have the source from contaminated environmental water by V. Cholerae. PMID- 10668269 TI - [Comparation of the antero-lateral and posterior approaches in primary total hip arthroplasty]. AB - Comparison between the antero-lateral and posterior approaches in primary total hip arthroplasty. In this retrospective study, 184 patients were enrolled, 95 submitted to the anterolateral (Watson jones) and 89 to the posterior approach (Moore) from June 1993 to June 1997. The outcomes assessed were perioperative data (operative time, hospital stay, time from surgery until hospital discharge, surgical bleeding and the need for blood transfusion), as well as late complications (deep venous thrombosis DVP, pulmonary embolism, periopheral nerve injury, prothesis instability and others). Both groups did not differ in terms of preoperative parameters. Those submitted to the posterior approach had shorter operative times (p < 0.001), as well as reduced bleeding (p < 0.05) and need for blood transfusion (p < 0.001) during surgery. The outcomes, such as late complications, had similar reduced frequency in both groups. The posterior approach has been successfully applied in our service and proves to be an excellent alternative surgical access to the total hip arthroplasty. PMID- 10668270 TI - [Amplifying right colectomy: place in the treatment of obstructive proximal left colon cancer]. AB - The results obtained about nineteen (19) patients operated by left colon cancer with variable grade obstruction have been analysed. Seventeen (17) patients operated due to obstructive left colon cancer situated: five (5) in distal transverse colon, other five (5) at splenic flexure and seven (7) in proximal descending colon but three of them with right synchronic neoplasias. The remaining two (2) that showed a cancer located at splenic flexure and the other one in proximal descending colon were reoperated three weeks later than a transverse colostomy had been performed owing to an obstructive condition. One patient had to be reoperated because a generalised peritonitis from a fistula with partial disruption on end to end ileo-colic anastomosis. Exteriorization of both ends was carried out with favourable evolution and subsequent reanastomosis. An exteriorized patient by splenic flexure cancer also had to be drained ten days later for a retroperitoneal abscess through a percutaneous puncture and a lesion grade 1 in lower pole of spleen was resolved with electrofulguration. No patient has showed invalidating diarrhea and all themselves have been stabilised with two or three stools daily about two month after surgery. Amplifying right colectomy is a safe procedure with low surgical morbimortality and take privileged place in the treatment of the patients undergoing synchronical neoplasias and/or carcinomas associated with polyps, specially in all those cases when a variable grade of obstruction have occurred. PMID- 10668271 TI - [Pancreaticogastrostomy]. AB - From 1991 we have carried out 19 duodenopancreatectomy and we have reconstructed with pancreatogastrostomy in 11 opportunities. The mortality was of 0% and the morbidity reached 52.94%. In this presentation we propose technical sample. PMID- 10668272 TI - [Value of the nuclear magnetic cholangio resonance in the study of the patient with jaundice]. AB - This report analyse the results about forty three (43) patients, thirty six (36) of which showed an extrahepatic obstructive biliary Syndrome was made evident by ultrasonography, five (5) with a cholecistolithiasis and doubtful history of jaundice were evaluated to carry out a video-surgery procedure and two (2) patients whom hepatic-yeyunostomy had been practiced, a control of anastomosis in postoperative period was required. Nuclear Magnetic Resonance and Operative Cholangiography findings were correlated and afterward with the anatomopathological studies when they arrived. In all cases the Nuclear Magnetic Cholangio Resonance (NMCR) let us prove the diagnosis of extrahepatic biliary obstruction determining with precision furthermore the topographical site of the lesion. Respecting the aetiology of obstruction, NMCR was accurate in 34 out of 36 cases (94.4%). In conclusion Cholangio-Resonance is an excellent diagnostic method to evaluate biliary ductal system including anatomic changes. However, there are some limitations yet in order to determine the aetiology of lesions about extrahepatic biliary via extremes. We emphasize its features such as non invasive, little operating dependent, and without morbimortality that become it as a method of choice to study the biliary via from a diagnostic viewpoint. PMID- 10668273 TI - [Restructuring and reorganization project in the public health area]. AB - The objective of this project is to produce substantial change as regards the reorganization, reconstruction and normalization of the hospital data to extra hospital data recording system from the Ministry of Health in the Province of Cordoba (Argentina). In order to incorporate socio-demographic factors in the various data forms so asa to produce efficiency upon the politic implemented. To achieve such aim, colwork on the different levels of execution, training the health team with respect to the identification of the population problems, and attempting to change of a demographic basis the current sanitary policies into policies of populations. PMID- 10668274 TI - [Cervical ectopic thymus]. AB - We present a case of ectopic thymus in an eight month old male baby, with a right lateral tumor of the neck. Ectopic thymus is a pathology rarely observed, its embryogenesis could explain its cervical localization. X Ray, ultrasonography, IRM, esophagoscophy and laryngoscophy may be helpful in the differential diagnosis with other tumors of the neck. Due to the fact that cystic lesions and neoplasis developments take place, the chosen treatment is the complete chirurgical extirpation. But at the absence of symptoms, no treatments is advisable because eventually the thymus spontaneously involutionates. PMID- 10668275 TI - Cytokine-like substance: origin and fate in Chagas' disease a new hypothesis about the local inflammatory reaction etiopathogenesis (experimental study in white mice) PMID- 10668276 TI - Orifice Diseases Project--experience of the "Hospital das Clinicas" University of Sao Paulo Medical Center in day-hospital of anorectal disease. AB - The treatment of malignant or benign colorectal pathologies that require more complex management are priorities in tertiary hospitals such as "Hospital das Clinicas" University of Sao Paulo Medical Center (HCFMUSP). Therefore, benign, uncomplicated orifice conditions are relegated to second place. The number of patients with hemorrhoids, perianal fistulas, fissures, condylomas and pilonidal cysts who seek treatment at the HFMUSP is very great, resulting in over-crowding in the outpatient clinics and a long waiting list for recommended surgical treatment (at times over 18 months). The authors describe the experience of the HCFMUSP over an eight-day period with day-hospital surgery in which 140 patients underwent surgery. Data was prospectively taken on the patients undergoing surgery for benign orifice pathologies including age, sex, diagnosis, surgery performed, immediate and late postoperative complications, and follow-up, 140 patients operated on over eight days were studied, 68 were males (48.75%) with ages ranging from 25 to 62 (mean 35.2 yrs.). Hemorrhoids was the most frequent condition encountered (82 hemorrhoidectomies, 58.6%), followed by perineal fistula (28 fistula repairs, 20.0%). The most common complication was headache secondary to rachianesthesia occurring in 9 patients (6.4%). One patient (0.7%) developed bleeding immediately PO that required reoperation. Mean follow-up was 104 days. Day-surgery characterized by quality care and low morbidity is feasible in tertiary public hospitals, permitting surgery for benign orifice pathologies on many patients within a short period of time. PMID- 10668277 TI - Effect of hyperbaric oxygen on the regeneration of experimental crush injuries of nerves. AB - Hyperbaric oxygen has been successfully used on treatment of acute ischemic injuries involving soft tissues and chronic injuries. In nerve crush injuries, the mechanisms involved are very similar to those found in ischemic injuries. Consequently, it is logical to hypothesize that hyperbaric oxygen should improve nerve repair, which is a critical step on functional recovery. In the present study, we created standard nerve crush injuries on sciatic nerves of rats, which underwent treatment with hyperbaric oxygen. Results were assessed by functional evaluation using walking-track analysis. The functional recovery indexes observed did not differ from control group. We concluded that hyperbaric oxygen therapy, in the schedule used, had no influence on functional recovery after nerve crush injuries. PMID- 10668278 TI - Rheumatic fever: a multicenter study in the state of Sao Paulo. Pediatric Committee--Sao Paulo Pediatric Rheumatology Society. AB - Rheumatic fever is still the most commonly seen rheumatic disease in Brazilian pediatric rheumatology clinics. It remains a significant health problem since subsequent cardiac sequelae represent one of the most important causes of chronic heart disease in children. We reviewed the clinical manifestations of rheumatic fever in 786 patients followed at seven pediatric rheumatology clinics in the state of Sao Paulo, Brazil. All patients were diagnosed according to revised Jones' criteria. Regarding major criteria, 396 (50.4%) children exhibited carditis, 453 (57.6%) polyarthritis, 274 (34.8%) chorea, 13 (1.6%) erythema marginatum, and 12 (1.5%) subcutaneous nodules. Valvular lesions documented by echocardiography in the absence of accompanying auscultatory findings were found in 144 (18.3%) patients. Migratory polyarthritis was observed in 290 (64.0%) patients with articular involvement. Documented previous streptococcal infection assessed by serum antistreptolysin (ASO) titers occurred in 531 (67.5%) patients. Even though prophylaxis with benzathine penicillin was recommended to all patients, recurrent attacks were observed in 147 (18.7%). We emphasize the high frequency of chorea, silent carditis and recurrences in our series as well as the variable clinical presentation of arthritis in rheumatic fever. Multicenter studies should be encouraged to improve our understanding of the clinical features of rheumatic diseases in children and adolescents. PMID- 10668279 TI - Urinary tract infection in full-term newborn infants: value of urine culture by bag specimen collection. AB - OBJECTIVE: To evaluate the efficacy of urine culture by bag specimen for the detection of neonatal urinary tract infection in full-term newborn infants. Retrospective study (1997) including full-term newborn infants having a positive urine culture (> 100,000 CFU/ml) by bag specimen collection. The urinary tract infection diagnosis was confirmed by positive urine culture (suprapubic bladder aspiration method). The select cases were divided into three groups, according to newborn infant age at the bag specimen collection: GI (< 48 h, n = 17), GII (48 h to 7 d, n = 35) and GIII (> 7 d, n = 9). Sixty one full-term newborn infants were studied (5.1% of total infants). The diagnosis was confirmed on 19/61 (31.1%) of full-term infants born alive. Distribution among the groups was: GI = 2/17 (11.8%), GII = 10/35 (28.6%), and GIII = 7/9 (77.7%). The most relevant clinical symptoms were: fever (GI--100%, GII--91.4%) and weight loss (GI--35.3%, GII- 45.7%). Urine culture results for specimens collected by suprapubic aspiration were: E. coli GI (100%), GII (40%) and GIII (28.6%), E. faecalis GI (30%), Staphylococcus coagulase-negative GII (20%) and GIII (42.8%), and Staphylococcus aureus GII (10%). Correlation between positive urine culture collection (bag specimen method) and urinary tract infection diagnosis, using relative risk analysis, produced the following results: GI = 0.30 (CI 95% 0.08-1.15), GII = 0.51 (CI 95% 0.25-1.06) and GIII = 3.31 (CI 95% 1.8-6.06). The most frequent urinary tract infection clinical signs in the first week were fever and weight loss, while non-specific symptomatology occurred later. E. coli was most frequent infectious agent, although from the 7th day of life, staphylococcus was noted. The urine culture (bag specimen method) was effective in detecting urinary tract infection only after the 7th day of life. PMID- 10668280 TI - Liver synthesis function in chronic asymptomatic or oligosymptomatic alcoholics: correlation with other liver tests. AB - Liver function and its correlation with bilirubin and hepatic enzymes were evaluated in 30 male chronic asymptomatic or oligosymptomatic alcoholics admitted into the psychiatric hospital for detoxification and treatment of alcoholism. Hypoalbuminemia, lowered prothrombin activity, hypotransferrinemia and hypofibrinogenemia were detected in 32%, 32%, 28%, and 24% of patients, respectively. Transferrin was elevated in 8%. Greater prevalence of hyperbilirubinemia was found in patients with lowered prothrombin activity, hypofibrinogenemia, or hypotransferrinemia. No correlation was found between serum bilirubin or aminotransferase levels and normal or elevated albumin levels, time or activity of prothrombin, and fibrinogen levels. Serum alkaline phosphatase was elevated in normoalbuminemics and gamma-glutamyltransferase in patients with lowered prothrombin activity. Hypoalbuminemia was associated with hypofibrinogenemia, hypotransferrinemia with elevated aspartate aminotransferase or gamma-glutamyltransferase, and hypertransferrinemia with elevation of alanine aminotransferase. These data indicated the occurrence of hepatic dysfunction due to liver damage caused directly by alcohol or by alcoholism-associated nutritional deficiencies. PMID- 10668281 TI - Congenital hyperthyroidism: autopsy report. AB - We report the autopsy of a stillborn fetus with congenital hyperthyroidism born to a mother with untreated Graves' disease, whose cause of death was congestive heart failure. The major findings concerned the skull, thyroid, heart, and placenta. The cranial sutures were closed, with overlapping skull bones. The thyroid was increased in volume and had intense blood congestion. Histological examination showed hyperactive follicles. The heart was enlarged and softened, with dilated cavities and hemorrhagic suffusions in the epicardium. The placenta had infarctions that involved at least 20% of its surface, and the vessels of the umbilical cord were fully exposed due to a decrease in Wharton's jelly. Hyperthyroidism was confirmed by the maternal clinical data, the fetal findings of exophthalmia, craniosynostosis, and goiter with signs of follicular hyperactivity. Craniosynostosis is caused by the anabolic action of thyroid hormones in bone formation during the initial stages of development. The delayed initiation of treatment in the present case contributed to the severity of fetal hyperthyroidism and consequent fetal death. PMID- 10668282 TI - [Acute myocardial infarction: the evolving knowledge about the disease]. AB - Important advances in the knowledge of pathophysiology and management of acute myocardial infarction have occurred lately. The results of large multicentric randomized trials allowed the introduction of new diagnostic techniques, patient stratification tools, drugs and treatment strategies to clinical practice. As a result, a significant reduction in mortality has been achieved. However there is a great variation in the adoption of the new diagnostic and therapeutic recommendations in different countries. In this issue of Revista Medica de Chile, a registry of patients with acute myocardial infarction treated in Chile between 1993 and 1995, is reported. The mean age of patients (62 +/- 12 years old), the proportion of males (74%) and the prevalence of risk factors is similar to that reported in series from developed countries. Similarly, 33% of patients received thrombolytic therapy, and the pattern of drug use was comparable. The 13.4% global mortality can be improved. An early consultation to health services when an acute myocardial infarction is suspected should be encouraged in the population. Likewise, treatment norms for health institutions should be devised. PMID- 10668283 TI - [Acute myocardial infarction in Chilean hospitals. Final results of the GEMI study]. AB - BACKGROUND: Acute myocardial infarction is the leading cause of death in Chile. AIM: To report the main features, hospital evolution, complications and pharmacological treatment of patients admitted to Chilean hospitals with the diagnosis of acute myocardial infarction. PATIENTS AND METHODS: Between 1993 and 1995, the GEMI group registered 2,957 patients admitted to 37 hospitals with the diagnosis of acute myocardial infarction. RESULTS: Mean age of patients was 62 +/ 2 years old and 74% were male. Forty six percent had a history of hypertension and 40% were smokers. During the first five days of admission, 93% of patients received aspirin, 95% received intravenous nitrates, 59% intravenous heparin, 56% oral nitrates, 37% beta blockers, 32% angiotensin-converting enzyme inhibitors, 33% thrombolytic agents, 29% antiarrhythmics and 23% calcium antagonists. Coronary angiograms were performed in 28% of patients, angioplasty in 9% and 8% were subjected to a coronary bypass. Global hospital mortality was 13.4% (19.5% in women and 11.1% in men, p < 0.001). CONCLUSIONS: This work gives a picture of myocardial infarction in Chilean hospitals. Pharmacological treatment is similar to that used abroad, but certainly it can be optimized. PMID- 10668284 TI - [Immunoglobulin rearrangement in the differential diagnosis of primary gastric lymphoma]. AB - BACKGROUND: The traditional methods to distinguish Chronic Follicular Gastritis and Primary Gastric Lymphoma do not allow an adequate definitive diagnosis in a significant number of cases. The molecular Biology diagnostic methods are based on the rearrangement of immunoglobulin genes. The polymerase chain reaction (PCR) specifically amplifies this rearrangement and allows molecular analysis of minimal tissue samples obtained with endoscopical biopsies. AIM: To test the usefulness of this PCR method in the differential diagnosis between Chronic Follicular Gastritis and Primary Gastric Lymphoma. MATERIAL AND METHODS: We analyzed the endoscopical biopsies of six Chronic Follicular Gastritis cases and eight surgically treated Primary Gastric Lymphoma cases, six with the correct diagnosis in the endoscopical biopsies and two with a diagnosis of Chronic Follicular Gastritis. RESULTS: A policlonal immunoglobulin rearrangement was found in the six cases with Chronic Follicular Gastritis. A monoclonal arrangement was found in 5 of 6 biopsies with the diagnosis of Primary Gastric Lymphoma. The same monoclonal rearrangement was observed in the two biopsies incorrectly diagnosed as Chronic Follicular Gastritis. CONCLUSIONS: PCR analysis of immunoglobulin rearrangement is a useful method in the differential diagnosis between Chronic Follicular Gastritis and Primary Gastric Lymphoma. PMID- 10668285 TI - [Use of a simple polymerase chain reaction combined with a heteroduplex mobility assay to characterize the non coding 5' region of hepatitis C virus]. AB - BACKGROUND: One of the most used methods for the characterization of hepatitis C virus strains is the use of a nested polymerase chain reaction (PCR) with a restriction fragment length polymorphism (RFLP) assay. Sometimes, RFLP results do not differentiate new strains. There are other more complex methods and only the sequencing of the PCR fragment allows a correct characterization of the strain. AIM: To report the detection of hepatitis C virus using a single PCR assay of the 5' non codifying region. MATERIAL AND METHODS: Thirty five serum samples coming from patients with chronic hepatitis or blood donors were assayed for hepatitis C virus. RESULTS: The reported method increases the PCR sensitivity through the combination of polyacrylamide gel electrophoresis and silver staining of amplified products. This allowed the semi quantitative estimation of viral load and the characterization of amplified products through their electrophoretic motility. These PCR products were used in a heteroduplex motility assay; allowing the discrimination between sequences of different genotypes. CONCLUSIONS: Heteroduplex assays can be used to characterize the 5' non codifying region of the hepatitis C virus for routine laboratory purposes. PMID- 10668286 TI - [Television publicity and food preferences of school age children of the metropolitan region]. AB - BACKGROUND: There is an alarming increase in the prevalence of child obesity in Chile. Lack of exercise and bad feeding habits strongly strongly contribute to the problem. AIM: To investigate the influence of television publicity on school age children food preferences. MATERIAL AND METHODS: A semi structured interview was applied to a representative sample of 786 school age children aged 6 to 11 years old, living in Metropolitan Santiago. Time watching television during week days and the attitude towards food and beverage commercials was analyzed and related to food preferences. RESULTS: Ninety nine percent of school age children watch television during week days and 20% watches more the three hours daily. Snack commercials such as those about potato chips, chocolates, cookies and ice cream, are preferred by 35% of children. Soda commercials are preferred by 33% and yoghurt commercials by 12%. Eighty five percent of children had money to buy food. Of these, 66% bought snacks, 15% bought sodas and 7% yoghurt. The same tendency was observed in school collations. CONCLUSIONS: The high percentage of children, watching television and the influence of commercials in their food preferences, requires an urgent educational strategy to promote healthy feeding habits. PMID- 10668287 TI - [High prevalence of undiagnosed primary hyperaldosteronism among patients with essential hypertension]. AB - BACKGROUND: Classically, primary hyperaldosteronism was diagnosed in no more than 1% of patients with hypertension, when hypokalemia was used as the screening test. However, numerous patients with primary hyperaldosteronism do not have hypokalemia and the disease remains undiagnosed. AIM: To assess the prevalence of normokalemic primary hyperaldosteronism among patients classified as having essential hypertension. PATIENTS AND METHODS: One hundred hypertensive patients with a blood pressure over 145/95 were studied. Plasma aldosterone and plasma renin activity were measured in all. A primary hyperaldosteronism was diagnosed when high aldosterone levels (over 16 ng/dl) and low plasma renin activity (below 0.5 ng/ml/h) coexisted in two blood tests or the aldosterone/plasma renin activity ratio was over 50. A probable primary hyperaldosteronism was diagnosed when the ratio was between 25 and 50 and these patients were subjected to a Fludrocortisone test to confirm the diagnosis. A dexametasone suppression test was done to discard glucocorticoid remediable aldosteronism. An adrenal TAC scan was done to all patients with primary hyperaldosteronism. RESULTS: A diagnosis of primary hyperaldosteronism was reached in ten patients. Seven had elevated aldosterone and low plasma renin activity. In three the diagnosis was confirmed with the fludrocortisone test. All ten patients had normal serum potassium levels. Dexametasone suppression test was positive in three patients, that normalized their blood pressure levels. Adrenal TAC scans showed an adenoma in one patient and hyperplasia in another. CONCLUSIONS: Primary hyperaldosteronism is more frequent than previously thought, it is overlooked when hypokalemia is used as the screening test and it can only be diagnosed measuring plasma aldosterone and renin activity. PMID- 10668288 TI - [Usefulness of the measurement of insulin-like growth factor (IGF-I) and IGF-1 binding protein-3 (IGFBP-3) for the diagnosis of growth hormone (GH) deficiency in children]. AB - BACKGROUND: The diagnosis of GH deficiency (GHD) is based upon the results of GH stimulation tests, which have several drawbacks. AIM: To evaluate the usefulness of IGF-1 and IGFBP-3 for the diagnosis of GHD in prepuberal children. MATERIAL AND METHODS: We measured IGF-I and IGFBP-3 in three group of subjects: I. GHD (n: 24), height < -2SD for age (Z score, average +/- SD: -4.2 +/- 1.2), growth velocity < p10 (3.4 +/- 1.0 cm/year) and peak GH level on two GH stimulation tests < 7 ng/ml (1.2 +/- 0.6 ng/ml); II. Short non-GHD (NGHD, n: 32), height of 2.7 +/- 0.9 SD for age, growth velocity < p 25 (3.9 +/- 1.2 cm/year), and peak GH level on two GH stimulation tests > 7 ng/ml (15.3 +/- 6.9 ng/ml), y III. Normal school children (n: 35) with normal heights (-0.17 +/- 0.12 SD) were studied as controls. RESULTS: IGF-1 and IGFBP-3 were significantly lower in GHD than in NGHD and controls (p < 0.001), and in NGHD than in C (p < 0.001). We defined the normal range of both proteins as +/- 2 SD of the mean of the control group. Using this criteria, IGF-I was low in 21/24 GHD, and in 12/32 NGHD. IGFBP-3 was low in 22/24 GHD, and in 6/32 NGHD. Only 1 GHD patient had both exams in the normal range, suggesting that he is probably NGHD. 4/32 of the NGHD and both exams below normal range, suggesting that they are probably GHD. CONCLUSIONS: IGF-1 and IGFBP 3 are important tools for the diagnosis of GHD. PMID- 10668289 TI - [Intravenous tissue plasminogen activator in the treatment of acute ischemic stroke: feasibility, safety, and efficiency in the 2 first years of the clinical practice]. AB - BACKGROUND: The only effective therapy for the treatment of acute ischemic stroke is the infusion of tissue plasminogen activator in the first three hours after the onset of symptoms. AIM: To report the experience with tissue plasminogen activator infusion in the treatment of acute ischemic stroke. PATIENTS AND METHODS: Ten males and 10 females, aged 52 to 85 years old with an acute ischemic stroke, admitted within 89 min after the onset of symptoms were studied. Tissue plasminogen activator was infused following the guidelines designed by the National Institute of Neurological Disorders and Stroke (NINDS). Patients were assessed according to Rankin scale after three months of follow up. RESULTS: All patients had normal CAT scans. The delay between the onset of symptoms and the infusion ranged from 75 to 180 min. One patient had a gastrointestinal bleeding due to a gastric ulcer and one patient had a fatal intracranial hemorrhage. After three months of follow up, 38% of patients had a good recuperation (Rankin 0 to 1), 33% had a mild to moderate disability (Rankin 2 or 3) and 14% had a moderate to severe disability (Rankin 4). There was a 15% mortality. CONCLUSIONS: This series show that treatment of acute ischemic stroke with tissue plasminogen activator is feasible and safe. The obtained results are similar to those reported abroad. PMID- 10668290 TI - [Hurthle cell carcinoma of the thyroid. A 41 years experience]. AB - BACKGROUND: Hurthle cells can be found in non malignant thyroidal diseases such as Basedow Graves and Hashimoto diseases. When Hurthle cells comprise more the 75% of cellularity, they become a neoplasm. There are malignant variants of these neoplasms, constituted by follicular and papillary Hurthle cell carcinomas. AIM: To report a 41 years experience with Hurthle cell carcinomas of the thyroid. MATERIAL AND METHODS: A retrospective review of all patients operated for a thyroid carcinoma and selection of those patients with Hurthle cell carcinoma of the thyroid. RESULTS: Twenty two patients (21 female, mean age 48 years old) with Hurthle cell carcinoma of the thyroid were selected. Total or near total thyroidectomy was the treatment of choice in 20 and 19 received 131I. Metastatic involvement of cervical lymph nodes was found in five patients and they underwent modified cervical lymph node dissection. During follow up, only one patient died of the disease. We did not find higher incidences of local recurrences, distant metastases or mortality rates, compared to well differentiated thyroid carcinomas. CONCLUSIONS: Hurthle cell carcinomas of the thyroid and well differentiated thyroid carcinomas have similar biological behaviors. Their treatment should be similar, including total or near total thyroidectomy plus modified cervical node dissection when there is lymph node involvement. Radioactive iodine therapy and suppressive levothyroxin therapy should follow. PMID- 10668291 TI - [Amiodarone induced retrobulbar neuritis. Clinical case]. AB - We report a 66 years old male, with an ophtalmologic history of long sightedness, admitted to the hospital due to paroxysmal atrial fibrillation crises in the context of a coronary heart disease. He was treated with i.v. amiodarone, receiving a total dose of 6 g in 72 hours. After the third day of treatment, the patient noticed a correction of his long sightedness and 24 h later, he complained of hlurred vision and orbital frontal headache. Visual field examination revealed a concentric retraction of visual field and a centrocecal scotoma in both eyes. Amiodarone was withdrawn and dexametasone treatment was begun. Three days after amiodarone discontinuation, sight improved and visual field returned to normal. Although retrobulbar neuritis has been associated to various drugs, amiodarone has not been considered as a possible agent. PMID- 10668292 TI - [Chronic atrial flutter: an infrequent manifestation of sick sinus syndrome. Clinical case]. AB - We report a 41 years old female, previously operated of an atrial septal defect, presenting with a persisting atrial flutter. Sinus node dysfunction became evident during an electrophysiological study at the moment of interrupting the flutter with electrical stimulation. The patient was treated with his bundle ablation and implantation of a definitive pacemaker. After one year of follow up, she is devoid of symptoms. PMID- 10668293 TI - [Sertoli-Leydig tumor. Clinical case]. AB - We report a 16 year old girl presenting with secondary amenorrhea, a history of voice coarsening, hirsutism and a body mass index of 35 kg/m2. Pelvic ultrasound and CT scans showed a retro uterine dense mass. She was operated and a left ovarian tumor was excised. Pathological examination disclosed a Sertoli-Leydig tumor. PMID- 10668294 TI - [Molecular biology and medicine: basic concepts]. AB - Through the advancements of molecular genetics, physicians and researchers are in an extraordinary period of study concerning the molecular basis of medicine. Molecular biology is making a tremendous impact on both diagnosis and treatment of diseases through the clinical introduction of molecular methods. These techniques, restricted for many years to basic biological research, include the polymerase chain reaction, DNA and protein electrophoresis, cloning of genes into viral or bacterial vectors and methods to rapidly sequence DNA and identify mutations. In this article the authors attempt to provide basic concepts on these themes for the non-trained physicians in order to help them to understand recent developments and foresee their future implications. PMID- 10668296 TI - [Efficacy indicators for the allocation of health resources]. AB - This review proposes an analytical method for the development of efficacy indicators, that will allow the integration of diverse technical criteria in the allocation of health resources. Indicators of epidemiological, clinical, organizational and economic efficiency were the four levels of conceptual approach integrated in the analytical framework. The different elements of each level are interrelated to compose an analytical perspective that can be used to guide the mechanisms of resource allocation based on technical criteria. This perspective allows the development of new relevant public health instruments, specially designed for the allocation of resources. PMID- 10668295 TI - [Adequacy of continuous ambulatory peritoneal dialysis in children]. AB - When the use of dialytical therapy is decided after a careful assessment of clinical and laboratory variables, the close supervision of the procedure, that allows a feedback between our indications and its clinical efficacy, is essential. The correct and routine use of validated adequacy tools such as Kt/V and the Peritoneal Equilibration Test (PET) is mandatory. We compare the adequacy figures for adult and pediatric populations, mentioning the Kt/V and PET values obtained in eight patients followed during 12 months in a Nephrology Unit. An initial Kt/V of 2.04 and of 2.14 after 12 months of procedure are values that adjust to the general recommendations discussed in this paper. According to PET results, this group of patients were classified as low average for ultrafiltration and high average for creatinine clearance. Based on the local experience and literature review, some recommendations are made for the management of peritoneal dialysis in children. PMID- 10668297 TI - [A medical history of Bernardo O'Higgins (1778-1842)]. AB - Bernardo O'Higgins was a very apprehensive individual regarding his health and ailments. This fact is clearly reflected in his letters, that provide valuable anamnestic data. During his youth, while living is Spain, he suffered of yellow fever and later in Chile, he probably had an acute phase of a rheumatic fever. Since his adolescence, he was affected by a chronic hlepharo-conjunctivitis. During the Chilean independence revolution, he suffered several battle wounds. The most severe was a shot that affected both his right arm and elbow (1818). While living in Peru (1823-1842) he suffered of dysentery and malaria. The latter was an endemic disease in the valleys of Peru. Being previously asymptomatic, he started experiencing extensional dyspnea, angor pectoris and syncopal episodes in 1840. At that time, physicians diagnosed a hypertrophic cardiomyopathy. Analyzing his symptoms and taking into account their short term evolution, the author concludes that they were a consequence of either an aortic stenosis or coronary insufficiency. These led him to a heart failure that was his immediate cause of death in 1842. PMID- 10668298 TI - [Organ transplantation in Chile]. PMID- 10668299 TI - Claudio Costa-Casaretto MD. (1914-1999) PMID- 10668300 TI - An empirical evaluation of the visual rightness theory of pictorial composition. AB - This research tested the visual rightness theory of pictorial composition's assertion that the induced organizational structure of a visually right (i.e., "good") design is perceptually salient and judged superior by anyone viewing it regardless of his or her training in the visual arts. Stimuli for Experiments 1 and 2 consisted of 16 reproductions of paintings by renowned artists and an experimentally reconstructed less-well-organized version of each art stimulus. It was found that design professionals (Experiment 2) were significantly more successful at detecting the original versions than were participants untrained in the visual arts (Experiment 1) (hit rates = 64% and 55%, respectively). In Experiment 3 participants replaced a major structural element removed from each of six pictures of the stimulus set at the location where they thought it appears in the original. A significant number of untrained participants and those with training in design theory were in agreement as to the location of each element within its pictorial field; the location chosen conformed to its compositional structure but not its actual location in the original. Findings demonstrate that the ability to detect the induced structural skeleton of a painting resulting from a visually right design does not require expert knowledge of design principles whereas the ability to discriminate between several articulation possibilities of the same composition does require formal training. PMID- 10668301 TI - Allocating attention in the visual field: the effects of cue type and target distractor confusability. AB - In the present study I compared the effect of colour and location cues on the selection of a target. Two questions were in focus. First, can both colour and location cues trigger and mediate attentional selection, and if they can, which of them is more effective? Second, does physical distinctiveness between a target and a distractor have to be taken into account? In two experiments, in which the most likely target colour or location was cued, both colour cues and arrow-like location cues produced cue-validity effects. The relative effectiveness of feature cues and location cues appeared, to some extent, to be dependent on target-distractor confusability. Taken together, the results support the post categorical filter theory, in which different cues can be used to trigger the feedback loop that is used to address relevant identity information via location. PMID- 10668302 TI - Response repetition benefits and costs. AB - In a wide variety of tasks, choice reaction time (RT) is reduced for repetitions of the previous response. However, when the task itself or a relevant physical feature that successive trials have in common changes, costs for response repetitions can be observed. In a series of three experiments it was investigated whether the repetition of a response results in costs if the stimulus category changes. Furthermore, it was asked whether there need to be informative physical task features that successive trials have in common to produce response repetition costs. In alternating runs, participants had to respond to either one of four symbols or one of the letters with a binary choice reaction: Results suggest that a change of stimulus category is a sufficient condition to produce response repetition costs. It is hypothesized that any change of a task feature that is part of the task representation participants adopt leads to a disruption of repetition-based facilitation and tends to facilitate a response alternation. PMID- 10668303 TI - The effects of eccentric head positions on leftward and rightward turns of a handle-bar. AB - We explored the structural constraints on concurrent movements and/or positions of the head and a steering device by means of a reaction-time task. In the first experiment, subjects had to respond rapidly to an imperative stimulus by way of rotating a handle-bar to the left or to the right and back to the central position while they maintained a left or right eccentric position of the head. Latency of the handle-bar responses did not depend on whether their initial directions were toward the eccentric head position or in the opposite direction, but kinematic characteristics did: iso-directional movements were of larger amplitude and longer duration until peak excursion. In the second experiment, the imperative stimuli for handle-bar rotations were presented at variable intervals after the head had been moved from the central to one of the eccentric positions and before its predictable return movement. Kinematic characteristics of the handle-bar rotations depended on the left and right eccentric head positions in the same way as in Experiment 1, but now iso-directional movements had a longer reaction time than movements in the direction of the forthcoming return movement of the head. These findings suggest that specifications of head-movement directions facilitate concurrent specifications of handle-bar rotations in the same direction and inhibit specifications of handle-bar rotations in the opposite direction, consistent with the notion of cross-talk during motor programming. PMID- 10668304 TI - Psychopathological aspects of cryptorchidism in children and adolescents. AB - In the male sexual and aggressive drives are mostly centered on the penis, whose real or fantasied features strongly affect the way children and adolescents build up their own personal and gender identity. In some clinical conditions a shift of genital centrality from penis to testicles is evident. The most frequent is abnormality in the descent of testicles, especially cryptorchidism, characterized by an arrested descent of one or two testicles that remain in the abdomen. The aim of this paper is to define mechanisms by which cryptorchidism increases psychological vulnerability. Time of diagnosis and treatment, restoration of genital integrity, personality stability and familial interactions are considered as elements affecting psychopathological outcome. Behavioral and psychological features in children and adolescents with cryptorchidism are reviewed. A case report of an adolescent with unilateral cryptorchidism is reported and discussed, as an example of pubertal distortion in bodily and gender identity. PMID- 10668305 TI - Clinical and developmental perspectives on adolescent coping. AB - Although the development of appropriate coping strategies has been understood as an essential element of healthy adjustment, few studies have demonstrated the link between coping and psychological development. Similarly, research on adolescents with behavioral problems has neglected coping as an important variable in understanding and treating these conditions. This study examines the relationships between psychological development, coping strategies and symptoms in a sample of 302 psychiatrically hospitalized adolescents, ages 12-16. Subjects completed the Adolescent Coping Orientation for Problem Strategies Questionnaire, (A-COPE), the Youth Self Report symptom checklist (YSR), and Loevinger's measure of ego development. Results showed that Avoidance and Ventilation were associated with increased behavior problems and lower levels of ego development. Problem solving and interpersonal strategies were associated with fewer symptoms and higher levels of development. Significant gender differences were found with girls using more interpersonal coping and boys using more physically active strategies. Gender differences were also found in the relationship of coping strategies to both symptomatic behavior and development. The results are discussed in the context of a developmental approach to adolescent psychopathology. PMID- 10668306 TI - Medication noncompliance in adolescents with psychiatric disorders. AB - The purpose of the study was to estimate prevalence of medication noncompliance among adolescents, following discharge from hospital. A second purpose was to identify predictors of such noncompliance. Seventy-one adolescents, who had been prescribed a medication during psychiatric hospitalization, were interviewed by telephone, 6-8 months post-hospitalization. Medication noncompliance was defined as discontinuing medication without the recommendation of the treating physician. Twenty-four subjects (33.8%) were noncompliant with medication. Age, race, gender, SES, diagnosis, type and number of medications, severity of depression, and family living arrangement did not predict noncompliance. We concluded that noncompliance with psychotropic medications was relatively common and difficult to predict in adolescents who had been hospitalized to a psychiatric inpatient unit; the majority of them suffered from depression. Clinicians should be aware that medication noncompliance may be common and a relatively unpredictable phenomenon. PMID- 10668307 TI - One-year-old infants of intrusive and withdrawn depressed mothers. AB - This study examined behaviors of intrusive/depressed versus withdrawn/depressed mothers and their one-year-old infants during a structured teaching interaction. Group comparisons revealed that intrusive/depressed mothers showed more positive responses, more demonstrating toys, and more physical guidance, and their infants demonstrated less toy manipulation. Withdrawn/depressed mothers maintained infant play more frequently and showed more restricted affect, and their infants demonstrated less affective behavior, both positive and negative. These findings suggested that exposure to depressed mothers' nonoptimal interaction styles represents different types of risk to infants' cognitive and affective development. PMID- 10668308 TI - Relations among child language skills, maternal socialization of emotion regulation, and child behavior problems. AB - Research has linked language delays in young children to behavior problems and risk for psychopathology. We hypothesized that low language skill would affect normal socialization of emotion regulation, which in turn would affect the development of behavior problems. Seventy-eight mother/preschool-age child pairs participated in two mildly frustrating situations. Parents of children with low verbal comprehension used more unexplained compliance demands than other parents. Further, children whose parents used more unexplained compliance demands used fewer cognitive and distraction strategies, and more instrumental strategies. Children's use of physical self-comforting was positively related to overall, internalizing, and externalizing behavior problems. Findings supported the original hypothesis. PMID- 10668309 TI - ECG of the month. Reading T leaves. Acute ischemic cardiac syndrome. PMID- 10668310 TI - Recurrent aphthous stomatitis. AB - Recurrent aphthous stomatitis is the most common oral mucosal disease in North America but it is commonly misdiagnosed and poorly understood. Pediatricians, internists, otolaryngologists, oral surgeons, and dentists may all be expected to treat this illness but little formal training in oral medicine may be offered to many of these health care professionals. This article reviews current evidence regarding etiology, pathogenesis, natural history, and treatment of this disorder. PMID- 10668311 TI - Radiology case of the month. Right upper quadrant pain and palpable mass. Hemangioma of the liver. PMID- 10668312 TI - The journal 150 & 100 years ago. January 1850 and 1900. PMID- 10668313 TI - How can genetics help in the management of obesity? AB - Obesity usually results from unwanted variations in metabolism. Inadequate neurotransmission, thermogenesis, or acylation underlie about 90% of cases. These are complex, weakly heritable, polygenic traits. Mutations in major gene loci cause another 5% of cases, and still another 5% of cases are due to gluttony. Careful observation can help define the type of obesity. All forms are associated with excess mortality and require lifelong episodic or continuous management. Management centers around diet, exercise, behavior therapy, and life-style counseling. Serotonin agonists and serotonin uptake inhibitors, as well as alternative therapies like phototherapy and 5-hydroxytryptophan are worthwhile for neurotransmitter inadequacy, except in children and pregnant women. When thermogenesis is inadequate, intake may be normal and weight reduction may require subnormal intake. Some degree of obesity may be required for optimal health in patients with inadequate acylation. In some Mendelian syndromes, obesity may balance a metabolic error, and weight reduction may restore metabolic imbalance. PMID- 10668314 TI - Dandy-Walker syndrome: presentation of the congenital malformation in an older patient. AB - Dandy-Walker syndrome, a congenital malformation of the hindbrain involving the cerebellum and the fourth ventricle, is a rare cranial abnormality that commonly occurs before the sixth or seventh week of development. It is usually diagnosed at birth or in early childhood; however, an occasional patient may first become symptomatic in adult life. A case of Dandy-Walker syndrome in a 58-year-old woman is reported because of the older age at presentation and relatively long asymptomatic period after birth. PMID- 10668315 TI - Lafayette's Family Practice Residency Program: practice patterns of graduates. PMID- 10668316 TI - Quadriceps sparing myopathy. AB - Magnetic resonance imaging (MRI) has been proven to be a useful tool in the evaluation of myopathy. Myopathic changes secondary to processes such as inflammatory disease, neuropathy, and neuromuscular disorders often involve several muscle groups. We describe a unique case of lower extremity myopathy with sparing of the quadriceps muscle group on MRI evaluation. PMID- 10668317 TI - [New discoveries on Alzheimer disease: new biochemical markers can hopefully improve diagnosis]. PMID- 10668318 TI - [Physicians and the spirit of the time 1930-1950. Situation on the labour market was the reason for prohibition of the import of physicians]. PMID- 10668319 TI - [Irrelevant about the risks of ocular laser interventions]. PMID- 10668320 TI - [Treatment of hepatitis C--recommendations by the specialty association are still valid]. PMID- 10668322 TI - [Private practitioners are not allowed to ask for the assistance of police]. PMID- 10668321 TI - [Clinical bacteriology and clinical virology are important parts of basic health care!]. PMID- 10668323 TI - [Toxic amblyopia and vitamin B12]. PMID- 10668324 TI - [Human blood is still the best for transfusion. But intensive research is going on to find substitutes]. PMID- 10668325 TI - [Future treatment of rheumatism. A compromise in the game between inflammation and infections]. PMID- 10668326 TI - [Reduced number of citizens per general practitioner. Increased support from the society to family practice in future]. PMID- 10668327 TI - [Goethe about medicine and his diseases: Yes, the soul of medicine is simple to understand. Laws of the great and the little world are studied--than everything is in the God's hands]. PMID- 10668328 TI - [Ancient medicine is based on nature philosophy. Alkmaion and Empedokles, philosophers and physicians seeking natural explanations]. PMID- 10668329 TI - [A project on "Hazardous use of alcohol". Patients receive a basis for their decisions concerning their alcohol habits]. PMID- 10668330 TI - [We manage better without nitrous oxide]. PMID- 10668331 TI - [Enough acid rain over psychiatry! The psychiatric society on the debate about psychiatry]. PMID- 10668332 TI - [The good psychiatric "asylums" as cleaning women of society]. PMID- 10668333 TI - [Projective tests and evaluations based on the psychodynamic theory are not reliable]. PMID- 10668334 TI - [A comment: better standard data are required. No resources for research]. PMID- 10668335 TI - [The histologist and anatomist Gustaf Retzius. He never received the Nobel Prize in spite of his twelve nominations]. PMID- 10668336 TI - [Mystery of the disappeared picture]. PMID- 10668337 TI - [A medical student of exactly 100 years ago. Battlefields and war prisoners' camps were his surgeries]. PMID- 10668339 TI - [Effect of exercise tolerance test on hemostasis in patients with and without coronary heart disease]. AB - BACKGROUND: Physical exercise leads to an elevated coagulation activity with a possibly disturbed hemostatic balance. Therefore patients with coronary heart disease have a potentially increased risk of thromboembolic events after a bicycle exercise tolerance test, that is frequently performed for diagnostic reasons. PATIENTS AND METHODS: Patients with angiographically known coronary heart disease (Group I: n = 49; age 59 years; male = 42, female = 7) were investigated in comparison to a healthy cohort (Group 2: n = 51; age 53 years; male = 44, female = 7) to study the influence of a standardized exercise tolerance test on hemostatic variables. Blood samples were taken before and after exercise. RESULTS: No significant changes were found for any investigated parameter between both groups. However, 3 parameters did change significantly within the groups: factor VIII rose in Group 1 from 132 to 156% and in Group 2 from 106 to 136% and the von Willebrand factor rose in Group 1 from 230 to 249% and in Group 2 from 228 to 247%. An elevated fibrinolytic potential was found with an increase of plasminogen-alpha 2-antiplasmin in Group 1 from 251 to 401 micrograms/l and in Group 2 from 247 to 350 micrograms/l. CONCLUSION: The findings underline the clinical presumption that exercise tolerance test does not increase the risk for thromboembolic complications in patients with coronary heart disease in comparison to patients without coronary heart disease, as long as the exercise tolerance test is performed in a standardized way and under aerobe conditions. PMID- 10668338 TI - [Economic evaluation of different treatment strategies in patients with stable angina pectoris or asymptomatic myocardial ischemia on basis of results from the Asymptomatic-Cardiac-Ischemia Pilot study (ACIP)]. AB - BACKGROUND: In patients with stable angina pectoris or silent myocardial ischemia, who had signs of ischemia in ECG during exercise, the Asymptomatic Cardiac Ischemia Pilot (ACIP) study compared 2 types of medication strategies (ischemia-guided and angina-guided) and a strategy of primary revascularization by PTCA or CABG. ACIP substantiated, after 2 years of observation, a clear advantage of the revascularization strategy compared to both drug strategies in terms of clinical effectiveness. This advantage is even more distinct in patients with very severe angiographic results. PATIENTS AND METHODS: This is a retrospective, incremental cost-effectiveness analysis from the perspective of the German third party payer (statutory sick funds) on the basis of the ACIP study. RESULTS: The direct costs of the revascularization strategy after 2 years are about 2 to 3 times higher than those of the drug therapies. The incremental cost-effectiveness of the ischemia-guided strategy versus an angina-guided strategy is DM 2,600 per life-year saved. Furthermore, the cost-effectiveness of a revascularization strategy versus an angina guided therapy is DM 15,100 per life-year saved. CONCLUSION: A primary revascularization strategy is cost effective in patients with stable coronary artery disease with proven myocardial ischemia and positive angiographic signs. PMID- 10668340 TI - [Drug therapy of endocrine neoplasms. Part I: Thyroid neoplasms, adrenal neoplasms and parathyroid neoplasms]. AB - BACKGROUND: The incidence of endocrine carcinomas is about 5.3 persons per 100,000 population. Most frequent are malignancies of the thyroid gland (about 89%). THERAPY: Because of low incidences and missing prospective studies as well as radiotherapy and chemotherapy resistance, general accepted therapy guidelines for endocrine carcinomas are still missing. Surgery and radionucleotide treatment is generally the first-line therapy. Hormonal active carcinomas can be additionally treated with special substances such as octreotide and mitotane. Chemotherapy is frequently not effective. Widely used substances are cyclophosphamide, cisplatin, doxorubicine, dacarbazine, vincristine and etoposide. This first part of the review will present medical therapies of thyroid carcinomas, adrenal carcinomas and parathyroid carcinomas. The second part in one of the next issues will focus on less frequent endocrine carcinomas of the gastrointestinal tract. PMID- 10668341 TI - [Original brands and generic preparations]. AB - BACKGROUND: Modern guidelines of drug approval aim at interchangeability of drugs containing the same active ingredients. Therapeutic equivalence of original and generic drugs is assumed as soon as bioequivalence is documented. For this to be accepted, first, pharmaceutical equivalence must prevail (the same amount of active substances in the same dosage forms) and, second, differences in bioavailabilities must not exceed certain limits. Drastic deviations from the original--not infrequent in the past--have become rare under the new sets of rules. CURRENT RELEVANCE: However, there is still room for sometimes stunning discrepancies between approved drugs, since the current procedures--mainly for economic reasons--do not cover some potentially relevant aspects: Usually, studies are performed on young, healthy, mostly male volunteers; possible effects of meals on bioavailability are not investigated and, after approval of a drug, maintenance of attested quality--as with all manufacturers--is not monitored. Moreover, the tolerated deviations from the bioavailability of the original drug are quite large; with certain substances, a change from the generic drug with the lowest bioavailability to that with the highest could mean transition from low efficacy to a toxic dose level. Documented examples include carbamazepine, phenytoin, levothyroxin, verapamil, and aspirin. CONCLUSIONS: In conclusion, even today's sophisticated rules do not suffice to cover all eventualities. In particular, drugs with a narrow therapeutic range require close scrutiny in product selection. Pertinent are drug documentation as well as the distinguishing features of the respective manufacturers, mainly scientific support and record of product reliability. Besides the sometimes insufficient official documentation, the internet has been gaining importance as a source of information. PMID- 10668342 TI - [Possible genetic causes for late complications of diabetes mellitus]. AB - BACKGROUND: Hyperglycemia occurs in every patient with diabetes mellitus. It is the most important factor in the development of diabetic complications. However, the onset, intensity and the progression of complications show large interindividual variations. Manifestation in families and the lack of complications in some diabetics with poor metabolic control indicate a genetic predisposition to develop diabetic complications like nephropathy, neuropathy and angiopathy. NEPHROPATHY: Diabetic nephropathy occurs only in 25 to 40% of the diabetic patients. Therefore a genetic risk factor for this complication is very likely. Various variations in genes like ACE-gene and angiotensinogen-gene have been described, which could be associated with the development of diabetic nephropathy. NEUROPATHY: Peripheral diabetic neuropathy occurs in up to 66% of all diabetics. Therefore and because of the possible pathological mechanisms genetic risk factors like variations in the Na/K-ATPase-gene and in the aldose reductase-gene are discussed. RETINOPATHY: An association between diabetic retinopathy and polymorphisms in the ACE-gene and the aldose reductase-gene seems very unlikely, because up to 75% of the diabetic patients suffer from retinopathy after 15 years of diabetes. MACROANGIOPATHY: A large number of studies show an association between diabetic macroangiopathy and genetic variations in the ACE gene (I/D-variant) and the paraoxonase-gene (2 isoforms). CONCLUSION: Based on the current evidence for associations of genetic markers with diabetic complications, the generation of an individual risk profile based on genetic markers seems to be possible. In addition to near euglycemia genetic markers could direct therapeutic strategies and lead to new therapeutic approaches. PMID- 10668343 TI - [Arthropathy in beta-thalassemia minor]. AB - CASE REPORT: A 25-year-old Turkish patient presented with a "swelling" of the ankles which now persisted for 5 years. He had previously been diagnosed as having HLA-B27 negative spondylarthropathy with peripheral joint involvement. The symptoms failed to respond to different therapies. Serological markers and radiological procedures revealed beta-thalassemia minor. DISCUSSION: Symptoms, diagnostic procedures and differential diagnostic options are discussed. PMID- 10668344 TI - [Ticlopidine-associated thrombotic thrombocytopenic purpura (Morbus Moschcowitz)]. AB - ANAMNESIS AND CLINICAL FINDINGS: A 75-year-old woman with a history of recurrent ischemic cerebral events was admitted with acute unspecific neurological symptoms and fever. EXAMINATION: Intracerebral hemorrhage due to hypertension and antithrombotic therapy with ticlopidine was ruled out with cranial computed tomography. Laboratory findings on admission included thrombocytopenia (12/nl), renal insufficiency (serum creatinine 1.6 mg/dl) and LDH elevation (1,218 U/l). The hemoglobin on admission was normal. THERAPY AND CLINICAL COURSE: In the presence of rapidly declining hemoglobin values and fragmentation of red cells thrombotic-thrombocytopenic purpura (TTP) was diagnosed and the patient received fresh frozen plasma. Shortly after the plasma infusion the patient's condition deteriorated rapidly showing clinical signs of an allergic shock. In the sequel of 24 to 48 hours the patient developed renal failure, severe anemia and the thrombocyte count fell to 5/nl. The patient was mechanically ventilated during the next 48 hours and needed intravenous catecholamines. Even after restoration of spontaneous respiration and cessation of pharmacological sedation the patient remained comatose. Cranial computed tomography on the fourth day after admission showed multiple infarction syndrome. The patient died on the ninth day after admission in status epilepticus which could not be stopped with pharmacological means. CONCLUSIONS: The combination of neurological symptoms, thrombocytopenia, fever, renal failure and hemolytic anemia in a patient taking ticlopidine points to a diagnosis of TTP. The high mortality of TTP can probably only be reduced by early plasmapheresis. PMID- 10668345 TI - [TSH-secreting pituitary adenoma: 16 years follow-up]. PMID- 10668346 TI - [German recommendations for health care economic evaluation studies. Revised version of the Hannover consensus. Hannover Consensus Group]. AB - Financial restrictions and stronger orientation towards outcomes increasingly demand rational decisions to be made about the use of resources in the health care system. Such decisions are the subject of medical, ethical and economic considerations. Management of the health care system requires medical and economic orientation both at the general level and with regard to the selection of suitable forms of care in hospital and medical practices. In this context, evaluative health economics can be a valuable decision-making aid. In order for the results of health economic evaluation studies to be validly interpreted, a minimum of standard methodology and sufficient transparency is required. To this end, recommendations were developed. They are intended to convey standard approaches, without unnecessarily constraining methodologic progress and scientific freedom. Ongoing refinement of the guidelines and adaptation of the current state of health economic research are desirable. PMID- 10668347 TI - [Sense and nonsense of post-authorization surveillance]. PMID- 10668348 TI - The rank-order consistency of personality traits from childhood to old age: a quantitative review of longitudinal studies. AB - The present study used meta-analytic techniques to test whether trait consistency maximizes and stabilizes at a specific period in the life course. From 152 longitudinal studies, 3,217 test-retest correlation coefficients were compiled. Meta-analytic estimates of mean population test-retest correlation coefficients showed that trait consistency increased from .31 in childhood to .54 during the college years, to .64 at age 30, and then reached a plateau around .74 between ages 50 and 70 when time interval was held constant at 6.7 years. Analysis of moderators of consistency showed that the longitudinal time interval had a negative relation to trait consistency and that temperament dimensions were less consistent than adult personality traits. PMID- 10668349 TI - Factors influencing racial comparisons of self-esteem: a quantitative review. AB - Research on racial comparisons of self-esteem was examined. Early research in this area, exemplified by the doll studies of racial preference, was viewed as demonstrating that Blacks have less self-regard than Whites. However, a meta analytic synthesis of 261 comparisons, based largely on self-esteem scales and involving more than half a million respondents, revealed higher scores for Black than for White children, adolescents, and young adults. This analysis further revealed that the direction and magnitude of racial differences are influenced by such demographic characteristics as participant age and socioeconomic status, as well as by characteristics of the measuring instruments. Many findings--for example, that the self-esteem advantage for Black respondents increases with age and is related to the sex composition of the sample--underscore the need for long term longitudinal studies of self-esteem development in male and female members of both racial groups. PMID- 10668350 TI - Evolution and proximate expression of human paternal investment. AB - In more than 95% of mammalian species, males provide little direct investment in the well-being of their offspring. Humans are one notable exception to this pattern and, to date, the factors that contributed to the evolution and the proximate expression of human paternal care are unexplained (T. H. Clutton-Brock, 1989). The nature, extent, and influence of human paternal investment on the physical and social well-being of children are reviewed in light of the social and ecological factors that are associated with paternal investment in other species. On the basis of this review, discussion of the evolution and proximate expression of human paternal investment is provided. PMID- 10668351 TI - Nonshared environment: a theoretical, methodological, and quantitative review. AB - When genetic similarity is controlled, siblings often appear no more alike than individuals selected at random from the population. Since R. Plomin and D. Daniels' seminal 1987 review, it has become widely accepted that the source of this dissimilarity is a variance component called nonshared environment. The authors review the conceptual foundations of nonshared environment, with emphasis on distinctions between components of environmental variance and causal properties of environmental events and between the effective and objective aspects of the environment. A statistical model of shared and nonshared environmental variables is developed. A quantitative review shows that measured nonshared environmental variables do not account for a substantial portion of the nonshared variability posited by biometric studies of behavior. Other explanations of the preponderance of nonshared environmental variability are suggested. PMID- 10668352 TI - Intuition: a social cognitive neuroscience approach. AB - This review proposes that implicit learning processes are the cognitive substrate of social intuition. This hypothesis is supported by (a) the conceptual correspondence between implicit learning and social intuition (nonverbal communication) and (b) a review of relevant neuropsychological (Huntington's and Parkinson's disease), neuroimaging, neurophysiological, and neuroanatomical data. It is concluded that the caudate and putamen, in the basal ganglia, are central components of both intuition and implicit learning, supporting the proposed relationship. Parallel, but distinct, processes of judgment and action are demonstrated at each of the social, cognitive, and neural levels of analysis. Additionally, explicit attempts to learn a sequence can interfere with implicit learning. The possible relevance of the computations of the basal ganglia to emotional appraisal, automatic evaluation, script processing, and decision making are discussed. PMID- 10668353 TI - Eyeblink classical conditioning: hippocampal formation is for neutral stimulus associations as cerebellum is for association-response. AB - Extensive evidence has been amassed that the cerebellum, hippocampus, and associated circuitry are activated during classical conditioning of the nictitating membrane/eyeblink response. In this article, the authors argue that the cerebellum is essential to all eyeblink classical conditioning paradigms. In addition, the septohippocampal system plays a critical role when the classical conditioning paradigm requires the formation of associations in addition to the simple association between the conditioned and unconditioned stimuli. When only a simple conditioned stimulus--unconditioned stimulus association is needed, the septohippocampal system has a more limited, modulatory role. The neutral stimulus association versus simple association-response distinction is one of the ways in which declarative or relational memory can be separated from nondeclarative or nonrelational memory in classical conditioning paradigms. PMID- 10668354 TI - Rhythm and pitch in music cognition. AB - Rhythm and pitch are the 2 primary dimensions of music. They are interesting psychologically because simple, well-defined units combine to form highly complex and varied patterns. This article brings together the major developments in research on how these dimensions are perceived and remembered, beginning with psychophysical results on time and pitch perception. Progressively larger units are considered, moving from basic psychological categories of temporal and frequency ratios, to pulse and scale, to metrical and tonal hierarchies, to the formation of musical rhythms and melodies, and finally to the cognitive representation of large-scale musical form. Interactions between the dimensions are considered, and major theoretical proposals are described. The article identifies various links between musical structure and perceptual and cognitive processes, suggesting psychological influences on how sounds are patterned in music. PMID- 10668355 TI - [The test for the evaluation speech perception and production]. AB - The aim of this paper is to present a Test to evaluate the speech perception and production in french profoundly hearing impaired children fitted with cochlear implant or traditional hearing aids. This test materials have been developed for children from 2 to 10 years old. This test fall into two basic categories known as speech perception and speech production. The assessment of the perceptual abilities through the use of the TEPPP is achieved across the hierarchy of perception which spans: detection, discrimination, identification, comprehension. The assessment of the speech production makes it possible to observe the speech production changes across three levels: phonetical, linguistical, intelligibility. The TEPPP, Test of Speech Perception and Production evaluation, enable us to observe if children, who receive a cochlear implant or a conventional hearing aids, obtain some degree of benefit in both speech perception and speech production. PMID- 10668356 TI - [The dynamics of the spoken language and parental guidance]. AB - The aim of parental guidance of deaf children is to allow the parents to understand and adapt to the unusual language development of their child. The authors propose a system of explanation of the control of language which will lead to the development of confidence, relaxation and spontaneity--the three determining factors--as well as to enlightenment; it will act as a point of reference during their introduction to the physiology of language, and as a guideline for the attitude of rehabilitation which needs to be adopted. PMID- 10668357 TI - [Value of the relative phonetogram (RP) for the evaluation of organic dysphonias]. AB - The phonetogram in a recognized element of voice evaluation, but its relation to perceptual voice quality is unclarified. The phonetograms area is easy to measure since the existence of efficient computer software. So information about frequency and intensity range can be united in one single parameter. The individual phonetogram area in relation to a gender- and training-specific normal value constitutes the "relative phonetogram (RP)". A prospective evaluation of the relative phonetogram was performed by means of a statistical analysis of its correlation to perceptual voice assessment (grade/rough/breathy) and to maximal phonation time. The acoustic parameters jitter, shimmer, SNR were examined in the same way, to allow for comparison of the RP's importance with the importance of common "objective" features in the identical group of patients. 114 patients with two subsets are included: 61 patients after partial laryngectomy (laser or conventional surgery), 53 patients with different glottic pathologies. The perceptive evaluation was done by a trained jury of an ENT-specialist and a speech therapist. The phonetogram and the maximal phonation time were measured by a trained medical student with regard of the examination references publicated by the Union of European Phoniatrics. The computer software for area measurement was MSImageProPlus, the one for sound analysis was Dr. Speech (Tiger Electronics). Statistical program: SPSS 8.0. RESULTS: The comparison between the two subsets of patients shows lower RPs for partial laryngectomy than for other patients in all degrees of hoarseness. In both subsets there is a correlation between RP and hoarseness values: the average values of RP differ significantly in dependence of grande. This is even more marked for the patients after laryngeal surgery. Furthermore high RPs are only present in patients with (relatively) high maximal phonation time, and mean RP correlates with maximal phonation time. A correlation between the parameters of sound analysis and the score of "grade" exists, but is not as marked as for the RP. CONCLUSION: The significance of the RP's mean value for subsets of 15-20 patients has been demonstrated. It is justified to interpret a certain variance of this parameter as difference in the degree of hoarseness. In this context, the importance of the mean RP is higher than the importance of jitter, shimmer, SNR (when measured with the above mentioned computer program, which allows no evaluation of parameter combinations). Therefore this parameter could be interesting for comparison of dysphonic patients, for instance after glottic cancer treatment. PMID- 10668358 TI - A study of vibrato: assessment by panel of judges compared to spectral voice analysis. AB - Experts in voice singing and voice training were asked to judge the vibrato of 30 singers (4 samples per singer: a sung [a] held without vibrato; a sung a[ held with vibrato; a self-selected passage and an imposed passage. In the first part, they ticked the type of oscillations (vibrato, straight tone, tremolo, quivering...). In the second part, they appraised various criteria. Intra-judge and inter-judge consistencies were determined. The subjective parameters were thereafter correlated with the measurements of six parameters of the MDVP (MultiDimentional Voice Program). The measurements were carried out on each task. Intra-judge consistency was good for only one judge (67-87% consistency). Since judge 2 and 3 were hardly reproducible (45-73%; 28-57%) measurement of the inter judge consistency was pointless. The results of judge 1 were correlated with the Fundamental Tremor Frequency Index, the jitter and the shimmer. PMID- 10668359 TI - The ultra-low resistance Groningen voice prosthesis: aerodynamic properties. AB - Post-laryngectomy voice rehabilitation using the low resistance (LR) Groningen voice prosthesis has over the past years provided good voice and speech results. The valve part of the prosthesis is largely responsible for the airflow resistance of the prosthesis. This study was performed to evaluate if by modifying the valve part of the LR Groningen prosthesis a lower airflow resistance in vitro could be achieved. Several prototypes with modified valves were tested. Based on the aerodynamic measurements the prosthesis with a single slit of 200 degrees in the 'hat' of the esophageal flange was selected for further evaluation and named the ultra-low resistance (ULR) Groningen voice prosthesis. Aerodynamic measurements were performed and showed the airflow resistance of the ULR Groningen voice prosthesis to be significantly lower than the resistance of the LR Groningen and the Provox voice prostheses. PMID- 10668360 TI - [Acoustic comparison of esophageal versus tracheoesophageal speech]. AB - The authors present results of an acoustic criterions analysis made next to 57 subjects, who had undergone total laryngectomy or total pharyngolaryngectomy, between 6 months and 3 years before. The study allows to assess, next to 29 tracheoesophageal voices and 30 oesophageal voices, the fundamental frequency, the vocal extensive, the maximum phonation time, and the study of pauses (number and time) in a reference sentence. The acoustic advantages of phonatory prothesis are confirmed, but the oesophageal voice knowledge must not be forgotten. PMID- 10668361 TI - [Long-term results of swallowing after horizontal sub-glottic laryngectomy]. AB - This is a retrospective study of 80 patients who have had a horizontal subglottic laryngectomy. The functional results were assessed by means of clinical findings at follow-up and a questionnaire. The mean time to decannulation was 32 days, and to removal of the nasogastric tube 25 days. 45% of patients had no fistula. Of the remaining 55%, there was a false passage for fluids in 90%. These fistulae remained patent for an average of 12 weeks. At one year and beyond, 40% of patients were on a normal diet, 33% were on ground food, 10% on a soft paste diet, 6% on a semi-liquid diet, and 10% were fed by tube. The factors militating towards poor function are age above 65 years, and previous radiotherapy. PMID- 10668362 TI - Studies on swallowing in patients operated by subtotal laryngectomy. AB - A series of 39 patients with problems of deglutition after total laryngectomy was studied by X-ray screening and manometry. The importance of the retropulsive movement of the tongue on the quality of deglutition has been demonstrated, as has the function of the superior oesophageal constrictor, the effect of a reduced relaxation time, and of the force of suction in reducing its quality. PMID- 10668363 TI - [Anatomy and anatomopathology of benign vocal cord lesions]. AB - After 30 years of experience comprising more than 3000 suspension laryngoscopies, we are putting forward a classification of the main benign lesions of the vocal cords. Among the acquired group, we make a distinction between those caused by vocal overuse and abuse and those with a cause within the vocal cord. Congenital lesions are certainly more common than is usually thought. The hypothesis of a congenital origin would certainly justify a larger prospective study. PMID- 10668364 TI - Flexible sigmoidoscopy revisited. PMID- 10668365 TI - Breaking the PPM contract. How to regain independence when PPM ownership no longer works. PMID- 10668366 TI - Unwinding the physician's employment: choosing a practice structure. PMID- 10668367 TI - Medical mistakes: decreasing the "oops" factor. PMID- 10668368 TI - Postoperative care--inattentive approach. PMID- 10668369 TI - Managed care preauthorization practices: implications for the emergency department. PMID- 10668370 TI - The hepatitis C epidemic in the hemophilia community. PMID- 10668371 TI - A patient with an acute viral syndrome. PMID- 10668372 TI - [Information for handling]. PMID- 10668373 TI - [Xenotransplantation]. PMID- 10668374 TI - [Vitamin A as an antidote against skin cancer?]. PMID- 10668375 TI - [Intracranial arachnoidal cysts--localization, gender and sidedness]. AB - The aim of the present study was to investigate whether data on location and distribution of intracranial cysts in a large patient population may explain why and how such cysts are formed. We investigated 123 patients with 129 intracranial cysts, consecutively admitted to the Department of Neurosurgery in Bergen 1988 97. Data were analyzed with regard to intracranial location and gender distribution. Cysts were much more commonly located in the temporal fossae than one would expect if the distribution were random; 68.1% of patients had temporal cysts. We suggest a theory that may explain how mal-development of the leptomeninges may contribute to the formation of cysts, and why such cysts are more common in the temporal fossae. Temporal cysts were significantly more frequent in males than in females (3.9:1), while cysts of other locations did not show preponderance for a specific gender. New in this study is the interesting connection between gender distribution and sidedness: the significant predominance of left-sided temporal cysts was found only in males. In patients with a unilateral temporal cyst, the left/right ratio was 2.0:1 (males 44 left and 20 right, females eight left and six right). We discuss whether the preponderance of left-sided temporal cysts in males can be explained by a gender specific developmental failure, as previously suggested for dyslexia. PMID- 10668376 TI - [Intracranial arachnoidal cysts--some neuropsychological experiences]. AB - Arachnoid cysts may cause neurological symptoms. The aim of this study was to investigate whether they also affect cognition. 31 patients (26 males and five females) underwent decompressive surgery for a symptomatic cyst in the left temporal fossa, with craniotomy and fenestration or with a cystosubdural shunt. The patients were tested for asymmetries in verbal perception and verbal memory before and after surgery, using dichotic listening techniques with different auditory stimuli presented simultaneously to the two ears. In the preoperative perception (dichotic listening-DL) test, the patients failed to show the normal advantage for auditory input to the right ear. After decompression, their DL performance normalised, as they now had a right ear advantage. In the preoperative dichotic memory test, the patients differed significantly from a normal reference group in two respects: they remembered fewer words, and they exhibited a clear left ear advantage, in contrast to the normal right ear advantage seen in the reference group. Postoperatively, overall memory performance was enhanced, and the preoperative superiority of the left ear had changed to a normal right ear advantage. The results indicate that arachnoid cysts in the left temporal fossa impair cognition, that neuropsychological tests are required to disclose such impairments, and that decompressive surgery improves cognition. PMID- 10668377 TI - [Transrectal specimens in recurrent rectal cancer]. AB - The majority of rectal cancer recurrences develop within the pelvis but outside the bowel. In clinical practice these tumours have been palpable in the pelvis for some time before a positive mucosal biopsy is obtained at the time when the cancer penetrates the mucosa of the neorectum or vagina. We present our experience with transrectal digitally guided fine needle aspiration cytology (FNAC) taken through intact mucosa or from intraluminal tumours. The sensitivity of the cytology was 0.88 versus 0.68 of the fine needle core biopsy (FNCB) specimens (p < 0.005); no false positive cancer diagnoses were reported. The simple and cost-effective digitally guided transrectal biopsy procedure may be adequate for an early morphological diagnosis, avoiding the more expensive CT or ultrasonically guided biopsies. It is safe and easy to take the biopsies and the complication rate is low. Surgical treatment for locally recurrent rectal cancer is justified. In combination with radiation therapy, it offers pain relief and improved quality of life. Survival is prolonged and cure can be achieved in up to one third of patients. PMID- 10668378 TI - [Hospital admissions due to obstructive lung diseases and pneumonia in two hospital districts]. AB - Admission rates for specific diseases in a defined district can be used as a coarse estimate of the need for health services. Such registration has not previously been conducted in the hospital districts of Bergen and Haugesund. We recorded all patients 16 years and older admitted to three hospitals for obstructive lung disease or pneumonia in the two hospital districts of Bergen and Haugesund in April to June 1997. A total of 438 patients were included in both districts, 246 with obstructive lung disease and 192 with pneumonia. The admission rate for obstructive lung disease was 359 per 100,000 inhabitants per year, for pneumonia 280 per 100,000 inhabitants per year. Median age was between 71 and 75 years for men and women for both diseases. 13% of patients with obstructive lung disease were readmitted to the hospital within 30 days. Obstructive lung disease and pneumonia are common causes of hospital admission, especially among older persons. The admission rates for both diseases combined were 639 per 100,000 adults per year in the hospital districts of Bergen and Haugesund. PMID- 10668379 TI - [Amputation or reconstruction of a circulatory compromised severely injured extremity?]. AB - 18 patients treated with primary or secondary amputations after severe lower limb open fractures were studied. All limbs had clinical signs of a compromised circulation at the primary evaluation. The various injuries are described and discussed with respect to the general guidelines for primary amputation. The Mangled Extremity Severity Score (MESS) and Nerve, Ischemia, Soft tissues, Skeletal, Shock, Age (NISSSA) scores were calculated. In view of the described injuries, primary amputation was indicated in ten patients according to the general recommendations, 11 patients according to NISSSA and 15 patients according to MESS. Delayed amputation leads t a significantly (p = 0.005) higher number of operative procedures than early amputation (9.2 vs. 2.9 treatments). The decision of whether to amputate or not should be based on sound clinical judgement, but injury scores such as MESS and NISSSA may be helpful. PMID- 10668380 TI - [Catheter-directed thrombolytic therapy--a current alternative to proximal deep venous thrombosis]. AB - Deep venous thrombosis extending into the iliac veins is associated with significant acute and late morbidity despite adequate conventional treatment with heparin and oral anticoagulants. The purpose of this paper is to focus on a multidisciplinary, aggressive approach with catheter-directed thrombolysis, in which a catheter with many side-holes is placed within the thrombus and thrombolytics infused. The aim is to eliminate the thrombus, to provide unobstructed venous drainage from the affected limb, and to prevent recurrent thrombosis. Total recanalisation of the iliofemoral segment was achieved in three of four treated patients, while partial lysis was obtained in one patient with symptoms for four weeks. Two weeks after discharge, two patients had no symptoms, while two had a moderate leg oedema. All of the three women used oral contraceptives at the time of thrombus formation; two had hereditary thrombophilia. Most probably, successful catheter-directed thrombolysis will reduce the incidence of post-thrombotic syndrome, but there are no long-term follow-up studies after such treatment. PMID- 10668381 TI - [Life-saving hemicraniectomy in acute massive brain infarction]. AB - Acute occlusion of the middle cerebral artery is an important cause of ischemic stroke. The resulting brain infarction is often very large, leading to massive brain oedema and intracranial hypertension. Despite intensive medical treatment, the mortality rate due to herniation and cerebral circulatory arrest remains very high. A Norwegian left-handed male, 28 years of age, developed signs of impending herniation following an acute right-side middle cerebral artery occlusion. When admitted to neurosurgical care 54 hours after the stroke, he was soporous, had a left-side hemiparalysis, conjugated deviation of the gaze and a mydriatic pupil on the right side. The intracranial pressure level was between 30 and 40 mm Hg. Following a right-side hemicraniectomy, the intracranial pressure fell to levels around 20 mm Hg. The bone flap was replaced four months later. One year after the stroke, the patient is fully independent of others, despite a left-side hemianopia and hemiparesis. Hemicraniectomy may be indicated in selected patients with impending herniation due to brain infarction. Intracranial pressure recordings are useful as an adjunct in the management of these patients. Our report seems to be the first account of decompressive hemicraniectomy performed in Norway for acute massive brain infarction. PMID- 10668382 TI - [Massive pulmonary embolism--echocardiographic diagnosis and thrombolytic therapy]. AB - Thrombolysis is widely accepted as the treatment of choice for acute massive life threatening pulmonary embolism. Several trials have shown that thrombolytic treatment has reduced morbidity and mortality in this condition, compared to heparin therapy. Rapid diagnosis and treatment start is mandatory for improving the prognosis. Clinical presentation, ECG, arterial blood gas analysis and D dimer are non-specific tests. Additional tests like ventilation/perfusion lung scan, spiral-computed tomography or pulmonary angiography are needed. In an emergency situation a non-invasive and bedside technique is preferred, and several studies have demonstrated the accuracy of echocardiography in pulmonary embolism. Acute right ventricular overload is indicated by different echocardiographic findings. Three cases with the echocardiographic method as the initial technique for the diagnosis of massive pulmonary embolism are described as well as the treatment consequence and practical administration of thrombolysis. Our patients had presenting features suggestive of massive pulmonary embolism, a clinical situation where an echocardiographic evaluation is appropriate. All presented with typical echocardiographic features. Thrombolytic treatment was initiated immediately after echocardiographic diagnosis. Different thrombolytic regimens were used. Echocardiography may be used as the initial imaging technique for the diagnosis of massive pulmonary embolism. The advantages are obvious, and thrombolytic treatment can be initiated without delay. PMID- 10668383 TI - [Decompression craniectomy--life-saving treatment in acute cerebral infarction]. AB - Massive cerebral infarction is often accompanied by early death secondary to transtentorial herniation. Decompressive hemicraniectomy has been suggested as a lifesaving procedure. We report the case of a 61 year old man who had an acute infarction in the distribution area of the right middle cerebral artery. Initially, he was awake and suffered from total left-sided hemiparalysis. Over the next two days, his level of consciousness deteriorated to a Glasgow Coma Scale score of 5. Intracranial pressure (ICP) monitoring was then established. Three days later, the ICP increased from 20 to 40 mm Hg. We performed a right sided decompressive hemicraniectomy, and the ICP was normalized immediately. Ten months after surgery the patient was at home and functioning with minimal assistance. He had moderate paresis of the left leg and was able to walk, but his left arm was paralytic. The presented case confirms that decompressive hemicraniectomy may prevent death and allow survival without severe disability in patients with massive cerebral infarction. PMID- 10668384 TI - [An elderly woman with constipation problems]. PMID- 10668385 TI - [Corneal transplantation]. AB - The first successful corneal transplantation was done in 1906, when the corneas of a recently deceased boy were transferred to a man blinded by an alkali burn. Since then, improved equipment, microsurgical technique and better methods for treating postoperative complications have greatly improved the prognosis of corneal grafting. Immunological reactions are less frequently seen after keratoplasty than after other types of transplantation, and the graft is also directly accessible for immunosuppressive treatment. Penetrating keratoplasty accounts for more than 95% of all corneal transplantations in Norway, the other procedures being lamellar grafting and epikeratophakia. While keratitis and macula corneae previously were the main indications for keratoplasty, keratoconus is at present the most frequent indication. The prognosis is poorer in eyes with extensive vascularization, lacking sensibility, insufficient tear secretion and marked symblepharon. Due to shortage of suitable donor tissue, the waiting lists for this type of ocular surgery are increasing. PMID- 10668386 TI - [Transplantation from animal to man]. AB - Because of a severe shortage of donor organs, organ transplantation therapy can only be offered to a fraction of all patients needing it. This has sparked an increasing interest in xenotransplantation. Transgenic pigs could potentially provide tissue and organs to thousands of patients on waiting lists world-wide. Xenografts may be used as bridges or as permanent alternatives to organ allotransplants. Clinical trials of transplantation of pig cells and tissues to treat diabetes and neurodegenerative disorders are in progress. The most striking immunologic obstacle to xenotransplantation of solid organs, that of hyperacute rejection, is close to being overcome. Next steps in research are to refine immunosuppression, improve our knowledge of physiology and to allow for small and strictly regulated clinical studies. However, the risk for the recipients from infectious agents, particularly retroviruses, and potential dissemination of these in the human population cannot be ignored and must be thoroughly investigated. Ethical, safety, and monitoring guidelines are needed to control the development of xenotransplantation. PMID- 10668387 TI - [Treatment of diabetes mellitus with transplantation of immunoprotected pancreatic islet tissue]. AB - Research in islet transplantation in the treatment of diabetes mellitus is making rapid progress. Vascularized pancreas allotransplantation is now an accepted treatment, but with major limitations. The shortage of human islets for allotransplantation requires new sources of insulin-producing tissue. Xenotransplantation is one such approach, though the procedure is limited by immunological rejection of the graft. Immunoisolation of insulin-producing graft in alginate capsules protects the transplant from immunological damage. The alginate capsule consists of a semipermeable membrane, which act as an immunological barrier between the graft and the host. In addition to exclude immunocells and humoral mediators of rejection, the membrane must be permeable to nutrients and toxic end products from the cell metabolism, and permit free diffusion of glucose and insulin. We may hope that this immunoisolating technique will establish prolonged survival of allo- or xenografted tissue without immunosuppressive treatment. PMID- 10668388 TI - [Abortion--family planning and a human right]. PMID- 10668389 TI - [Improved routines in connection with thrombolytic therapy in myocardial infarction]. PMID- 10668390 TI - [Clinical significance of antibodies against red blood cells]. PMID- 10668391 TI - [Ethical problems at the end of life]. PMID- 10668392 TI - [Family practice--professional content]. PMID- 10668393 TI - Protein folding and deficiencies caused by dominant-negative mutants of hormones. AB - Protein folding and transport in the secretory pathway of cells is a controlled process, facilitated by chaperones. Proteins that do not fold well elicit several different programmed responses from the cells. A comparison of mutants of growth hormone that result in growth hormone deficiency suggests that cells do not respond in the same way to all growth hormone mutants that cannot fold, because some mutants are dominant and some are recessive. Causes for autosomal dominant hormone deficiencies include accumulation of toxic or dysfunctional forms, competition for chaperones important for folding or transport, induction of protein degradation in the endoplasmic reticulum, or long-term responses of the cells to synthesis of proteins that do not fold that decrease hormone synthesis or cell viability. PMID- 10668394 TI - Structural aspects of the G-protein receptor, rhodopsin. PMID- 10668395 TI - Molecular modeling of mammalian cytochromes P450: application to study enzyme function. AB - Cytochromes P450 are important heme-containing enzymes that catalyze the oxidation of a vast array of endogenous and exogenous compounds, including drugs and carcinogens. One of the more successful approaches to study P450 function involves molecular modeling. Because none of the mammalian P450s have been crystallized, a number of homology models have been constructed based on the structures of known bacterial P450s. Molecular models, generated using molecular replacement or distance geometry methods, can be used to dock substrates and/or inhibitors in the active site to explain various aspects of enzyme function. The majority of modeling research has dealt with enzyme-substrate interactions in the active site. The analysis of these interactions has helped us to better understand the mechanism of P450 catalysis and provided the structural basis for the regio- and stereospecificity of substrate oxidation as well as susceptibility to inhibition or inactivation. The models have been utilized to identify and/or confirm key residues and to rationally interpret experimental data. The alteration in activity in a mutant P450 can be related to changes in enzyme substrate/inhibitor interactions, such as the removal or appearance of van der Waals overlaps or changes in compound mobility. Homology models can also help to analyze P450-redox partner interactions and identify critical determinants of protein stability. We can expect further development of molecular modeling methods and their increasing contribution into research on P450 function as an integral part of a combined theoretical-experimental approach. PMID- 10668396 TI - Prostaglandin D synthase: structure and function. AB - Prostaglandin (PG) D synthase catalyzes the isomerization of PGH2, a common precursor of various prostanoids, to produce PGD2 in the presence of sulfhydryl compounds. PGD2 induces sleep, regulates nociception, inhibits platelet aggregation, acts as an allergic mediator, and is further converted to 9 alpha, 11 beta-PGF2 or the J series of prostanoids, such as PGJ2, delta 12-PGJ2, and 15 deoxy-delta 12,14-PGJ2. We have purified two distinct types of PGD synthase; one is the lipocalin-type enzyme and the other is the hematopoietic enzyme. We isolated the cDNA and the gene for each enzyme and determined the tissue distribution profile and the cellular localization in several animal species. Lipocalin-type PGD synthase is localized in the central nervous system and male genital organs of various mammals and the human heart and is secreted into cerebrospinal fluid, seminal plasma, and plasma, respectively. The human enzyme was identified as beta-trace, which is a major protein in human cerebrospinal fluid. This enzyme is considered to be a dual-function protein; it acts as a PGD2 producing enzyme and also as a lipophilic ligand-binding protein, because the enzyme binds retinoids, thyroids, and bile pigments, with high affinities. Hematopoietic PGD synthase is widely distributed in the peripheral tissues and localized in the antigen-presenting cells, mast cells, and megakaryocytes. The hematopoietic enzyme is the first recognized vertebrate homolog of the sigma class of glutathione S-transferase. X-ray crystallographic analyses and generation of gene-knockout and transgenic mice for each enzyme have been performed. PMID- 10668397 TI - Steroid dehydrogenase structures, mechanism of action, and disease. AB - Steroid dehydrogenase enzymes influence mammalian reproduction, hypertension, neoplasia, and digestion. The three-dimensional structures of steroid dehydrogenase enzymes reveal the position of the catalytic triad, a possible mechanism of keto-hydroxyl interconversion, a molecular mechanism of inhibition, and the basis for selectivity. Glycyrrhizic acid, the active ingredient in licorice, and its metabolite carbenoxolone are potent inhibitors of human 11 beta hydroxysteroid dehydrogenase and bacterial 3 alpha, 20 beta-hydroxysteroid dehydrogenase (3 alpha, 20 beta-HSD). The three-dimensional structure of the 3 alpha, 20 beta-HSD carbenoxolone complex unequivocally verifies the postulated active site of the enzyme, shows that inhibition is a result of direct competition with the substrate for binding, and provides a plausible model for the mechanism of inhibition of 11 beta-hydroxysteroid dehydrogenase by carbenoxolone. The structure of the ternary complex of human 17 beta hydroxysteroid dehydrogenase type 1 (17 beta-HSD) with the cofactor NADP+ and the antiestrogen equilin reveals the details of binding of an inhibitor in the active site of the enzyme and the possible roles of various amino acids in the catalytic cleft. The short-chain dehydrogenase reductase (SDR) family includes these steroid dehydrogenase enzymes and more than 60 other proteins from human, mammalian, insect, and bacterial sources. Most members of the family contain the tyrosine and lysine of the catalytic triad in a YxxxK sequence. X-ray crystal structures of 13 members of the family have been completed. When the alpha-carbon backbone of the cofactor binding domains of the structures are superimposed, the conserved residues are at the core of the structure and in the cofactor binding domain, but not in the substrate binding pocket. PMID- 10668398 TI - Structure-based inhibitor design. AB - Time and costs associated with the discovery of new drugs have been significantly reduced by enzyme structure-based approaches to the discovery of new chemotherapeutic agents. However, fundamental components of the overall approach continue to rely on technologies which, by their nature, involve relatively random processes (i.e., combinatorial chemistry and high-throughput screening). Thus, the efficiency of the drug discovery process potentially could be further improved through better use of structural information. In this regard, three dimensional structures of enzymes are now being solved at high resolution and/or in conformations that provide data that should be more useful for inhibitor design or discovery. Scientists are beginning to appreciate the importance of water as a possible competitor of inhibitors for binding to target enzymes. New computational algorithms are improving the efficiency of identifying flexible inhibitors from among the large numbers of compounds in chemical databases. Also, tools of molecular genetics together with structures of target enzymes are likely to be used more frequently in dealing with the development of resistance to novel chemotherapeutic agents. Instead of detailing success stories in structure-based drug discovery, the following article considers how future efforts to discover or design new drugs may increasingly rely on information about molecular targets and less on data acquired via approaches involving random methodologies. PMID- 10668399 TI - Protein folding using contact maps. AB - We discuss the problem of representations of protein structure and give the definition of contact maps. We present a method to obtain a three-dimensional polypeptide conformation from a contact map. We also explain how to deal with the case of nonphysical contact maps. We describe a stochastic method to perform dynamics in contact map space. We explain how the motion is restricted to physical regions of the space. First, we introduce the exact free energy of a contact map and discuss two simple approximations to it. Second, we present a method to derive energy parameters based on perception learning. We prove in an extensive number of situations that the pairwise contact approximation both when alone and when supplemented with a hydrophobic term is unsuitable for stabilizing proteins' native states. PMID- 10668400 TI - HIV protease: enzyme function and drug resistance. AB - HIV protease is responsible for processing of the gag and gag-pol polyproteins during virion maturation. The activity of this enzyme is essential for virus infectivity, rendering the protein a major therapeutic target for AIDS treatment. This articles reviews the biochemical and biophysical properties of the enzyme. The clinical and in vitro observations of resistance to protease inhibitors are discussed from the perspective of drug resistance mechanisms of HIV protease mutants. PMID- 10668401 TI - Ultrastructure of cells undergoing apoptosis. PMID- 10668402 TI - Role of cytoskeleton in apoptosis. AB - Apoptosis is a form of cell death that takes place under physiologic conditions, and plays a key role in the control of biological processes such as embryonic development, tissue remodelation and renewal, or regulation of cell populations. Since its discovery in the early 1970s, there have been many relevant advances in the knowledge of the biochemical and molecular events involved in apoptosis. However, although the apoptotic process was defined on the basis of morphologic observations, only recently have we started to elucidate the molecular mechanisms that drive the structural changes observed in cells undergoing apoptosis. The article reviews current knowledge about the implications of cytoskeleton components (microfilaments, intermediate filaments, microtubules, and other cytoskeleton-related proteins) in the dynamics of apoptosis. PMID- 10668403 TI - Dietary antioxidants and cardiovascular disease. PMID- 10668404 TI - Melatonin receptors and ligands. AB - The goal of the article is to provide a clearer understanding of how melatonin and its related analogs interact with melatonin receptors with the hope of developing important tools and agents of significant clinical and scientific importance. The review provides a compilation of the currently published melatonergic ligands and their relative affinities for melatonin receptors and discusses the importance of developing reversible, high-affinity, and subtype selective melatonin receptor antagonists. In addition, the review discusses the utility of developing high-affinity charged melatonergic ligands and irreversible ligands. Finally, the review discusses some of the problems associated with the current models used to study receptor pharmacology and function. As the availability of tools increases in the melatonin receptor field, a great body of knowledge is also gained about the structure of the melatonin receptor and the role that specific melatonin receptor subtypes have in physiologic processes. Further design, synthesis, and application of melatonergic ligands will lead us to a clearer understanding of the role that melatonin and its receptors play in humans. PMID- 10668405 TI - Vitamin K-dependent proteins. AB - Vitamin K is required for the synthesis of gamma-carboxyglutamate (Gla) during postribosomal protein modification. Substrates include blood clotting proteins, bone proteins, cell signaling, and receptor proteins. In addition, Gla is a component of short toxin peptides from the marine snail Conus. Studies of structure-function relationships are the most advanced for the blood coagulation proteins. Reviews of vitamin K action and blood coagulation are presented. Special focus is on the structure-function role of Gla in blood coagulation and the impact of this amino acid on enzyme reaction kinetics. This amino acid forms calcium and membrane binding sites for these proteins. Two proposed mechanisms of protein-membrane attachment are reviewed. One involves membrane attachment by protein insertion into the hydrocarbon region of the membrane, while another considers attachment by specific interactions with phospholipid head groups. Membrane attachment generates the potential for several forms of nonclassical enzyme kinetic behaviors, all of which have been observed in vitro. For example, the reaction may be limited by properties of the enzyme active site, a condition that allows use of classic steady-state enzyme kinetic parameters. However, the reaction may be limited by substrate binding to the membrane, by substrate flux through solution, and/or by solvent flow rates across the membrane surface. These states provide special mechanisms that are not anticipated by classical steady state kinetic derivations. They may be used to regulate coagulation in vivo. Overall, vitamin K research spans the spectrum of biological research and experience. Exciting new ideas and findings continue to emanate from vitamin K related research. PMID- 10668406 TI - Steroid/nuclear receptor coactivators. AB - In higher eukaryotes, steroids/thyroid hormones and many lipophilic compounds regulate cellular physiology through binding to the steroid/nuclear receptor proteins. Steroid/nuclear receptors are ligand-dependent transcriptional activators that can stimulate gene expression. This transcriptional activation plays a pivotal role in hormone-regulated physiological and pharmacological responses. In recent years, several steroid/nuclear receptor cofactors have been identified and found to interact with the receptor and modulate its transcriptional activity. Among these cofactors, a family of three co-activators has been the focus of intense studies. Although gaps remain, progress has been made in understanding how a given co-activator interacts with the receptor and promotes transcriptional activation. We are beginning to understand co-activator action; for instance, several studies have established the molecular basis of antagonism by anti-hormones and the connection of co-activators with human cancers. PMID- 10668407 TI - Thyroid hormone receptor, v-ErbA, and chromatin. AB - The thyroid hormone receptor and the highly related viral oncoprotein v-erbA are found exclusively in the nucleus as stable constituents of chromatin. Unlike most transcriptional regulators, the thyroid hormone receptor binds with comparable affinity to naked and nucleosomal DNA. In vitro reconstitution experiments and in vivo genomic footprinting have delineated the chromatin structural features that facilitate association with the receptor. Chromatin bound thyroid hormone receptor and v-erbA generate Dnase I hypersensitive sites independent of ligand. The unliganded thyroid hormone receptor and v-erbA associate with a corepressor complex containing NCoR, SIN3, and histone deacetylase. The enzymatic activity of the deacetylase and a chromatin environment are essential for the dominant repression of transcription by both the unliganded thyroid hormone receptor and v erbA. In the presence of ligand, the thyroid hormone receptor undergoes a conformational change that weakens interactions with the corepressor complex while facilitating the recruitment of transcriptional coactivators such as p300 and PCAF possessing histone acetyltransferase activity. The ligand-bound thyroid hormone receptor directs chromatin disruption events in addition to histone acetylation. Thus, the thyroid hormone receptor and v-erbA make very effective use of their stable association with chromatin and their capacity to alter the chromatin environment as a major component of the transcription regulation process. This system provides an exceptionally useful paradigm for investigating the structural and functional consequences of targeted chromatin modification. PMID- 10668408 TI - U.S. Food and Drug Administration: future of new product strategies and FDA priorities. PMID- 10668409 TI - Neuroprotection against cerebral ischemia. A review of animal studies and correlation with human trial results. PMID- 10668410 TI - The pivotal role of iron in NF-kappa B activation and nigrostriatal dopaminergic neurodegeneration. Prospects for neuroprotection in Parkinson's disease with iron chelators. AB - R-Apomorphine (APO) the catechol-derived dopamine D1-D2 receptor agonist has been shown to be highly potent iron chelator and radical scavenger and inhibitor of membrane lipid peroxidation in vitro, in vivo and in cell culture employing PC12 cells. Its potency has been compared to the prototype iron chelator desferrioxamine (desferal), dopamine, nifedipine and dopamine D2 receptor agonists, bromocriptine, lisuride, pergolide and pramipexole. APO also inhibits brain and mitochondrial protein oxidation. In vivo APO protects against MPTP (N methyl-4-phenyl-1,2,3,6-tetrahydropyridine)- induced striatal dopaminergic neurodegeneration in C57 black mice with as low as 5 mg/kg. APO is a reversible competitive inhibitor of monoamine oxidase (MAO) A and B with IC50 values of 93 and 214 uM, respectively. The iron chelating and radical scavenging actions of desferal and APO explains their ability to inhibit iron and 6-hydroxydopamine (6 OHDA)-induced neurodegeneration and activation of redox-sensitive transcription factor NF-kappa B and the subsequent transactivation of promoters of genes involved in inflammatory cytokines. Iron is thought to play a pivotal role in neurodegeneration, and APO may be an ideal drug to investigate neuroprotection in Parkinson's disease where iron and oxidative stress have been implicated in the pathogenesis of nigrostriatal dopamine neuron degeneration. PMID- 10668411 TI - Hypothermia-induced ischemic tolerance. AB - Delayed resistance to ischemic injury can be induced by a variety of conditioning stimuli. This phenomenon, known as delayed ischemic tolerance, is initiated over several hours or a day, and can persist for up to a week or more. The present paper describes recent experiments in which transient hypothermia was used as a conditioning stimulus to induce ischemic tolerance. A brief period of hypothermia administered 6 to 48 hours prior to focal ischemia reduces subsequent cerebral infarction. Hypothermia-induced ischemic tolerance is reversed by 7 days postconditioning, and is blocked by the protein synthesis inhibitor anisomycin. Electrophysiological studies utilizing in vitro brain slices demonstrate that hypoxic damage to synaptic responses is reduced in slices prepared from hypothermia-preconditioned animals. Taken together, these findings indicate that transient hypothermia induces tolerance in the brain parenchyma, and that increased expression of one or more gene products contributes to this phenomenon. Inasmuch as hypothermia is already an approved clinical procedure for intraischemic and postischemic therapy, it is possible that hypothermia could provide a clinically useful conditioning stimulus for limiting injury elicited by anticipated periods of ischemia. PMID- 10668412 TI - A double-blind, placebo-controlled study of the safety, tolerability and pharmacokinetics of CP-101,606 in patients with a mild or moderate traumatic brain injury. AB - CP-101,606 is a postsynaptic antagonist of the glutamate-mediated NR2B subunit of the N-methyl-D-aspartate (NMDA) receptor. When administered intravenously (i.v.) at the time of injury, CP-101,606 is neuroprotective in animal models of traumatic brain injury (TBI) and ischemia. Minimal adverse effects have been observed in normal human volunteers given i.v. doses of up to 3 mg/kg/hr for 72 hours. The objective of the present clinical trial was to assess the safety, pharmacokinetics, and tolerability of CP-101,606 infused for various times in patients who had suffered either an acute moderate or mild TBI (Glasgow Coma Score 9-14) or hemorrhagic stroke. Patients began receiving treatment within 12 hours of brain injury. A total of 53 subjects (45 with TBI and 8 with stroke) were randomized in a double-blind fashion to receive CP-101,606 or placebo (4 drug: 1 placebo). Drug/placebo was administered by i.v. infusion (0.75 mg/kg/hr) for 2 hours and then stopped (n = 25) or continued for 22 hours (n = 4) or 70 hours (n = 24) at a rate of 0.37 mg/kg/hr. Mean plasma drug concentrations were well above the predicted therapeutic concentration of 200 ng/ml within two hours of initiating treatment and were sustained as long as drug was infused. All the patients tolerated their drug/placebo treatment, and there were no clinically significant cardiovascular or hematological abnormalities in either group. A Neurobehavioral Rating Scale, used to detect personality changes and behavioral disturbances, indicated that all subjects showed an improvement from their postinjury, predosing baseline but did not significantly differ from each other with respect to type of head injury and/or treatment with drug or placebo. Modified Kurtzke Scoring also showed a similar pattern of improvement irrespective of type of head injury or drug/placebo treatment. This study suggests that CP-101,606, infused for up to 72 hours has no psychotropic effects and is well-tolerated in patients who have sustained a mild or moderate TBI or hemorrhagic stroke. PMID- 10668413 TI - An open-label study of CP-101,606 in subjects with a severe traumatic head injury or spontaneous intracerebral hemorrhage. AB - CP-101,606 is a postsynaptic antagonist of N-methyl-D-aspartate (NMDA) receptors bearing the NR2B subunit. When administered intravenously (i.v.), it decreases the effects of traumatic brain injury (TBI) and focal ischemia in animal models. Therapeutic plasma concentrations (200 ng/ml) in animals, have been well tolerated in healthy human volunteers. The purpose of the present dose escalation study was to assess the safety, tolerability, and pharmacokinetics of CP-101,606 in subjects who had suffered either an acute severe TBI (Glasgow Coma Scale 3-8) or spontaneous intracerebral hemorrhage. Thirty patients, 20 with a TBI and 10 with a stroke, were enrolled in the trial and began receiving an i.v. infusion of CP-101,606 for 2 hours, 24 hours, or 72 hours within 12 hours of brain injury. For the first two hours, the drug was given a rate of 0.75 mg/kg/hr and then stopped (n = 17) or continued for 22 (n = 2) or 70 hours (n = 11) at 0.37 mg/kg/hr. Plasma and cerebrospinal fluid (CSF) were collected at serial times during and after treatment. There were no consistent changes in blood pressure or pulse nor any clinically significant hematological or electrocardiogram (ECG) abnormalities attributable to CP-101,606. No adverse events or behavioral changes were considered to be related to the drug. Plasma concentrations of CP-101,606 over 200 ng/ml were rapidly achieved in the blood and CSF within two hours and were sustained there as long as the drug was infused. CSF concentrations were slightly higher than that in plasma by the end of infusion suggesting good penetration of CP-101,606 into the CSF. Outcome in the severe TBI patients, as measured by the Glasgow Outcome Score at six months, suggested that a two-hour infusion yielded a range of scores similar to contemporary patients with a severe TBI treated at our hospital while the outcomes of the patients treated with either a 24- or 72-hour infusion were better on average. Thus, these results indicate that CP-101,606 infused for up to 72 hours is well tolerated, penetrates the CSF and brain, and may improve outcome in the brain-injured patient. PMID- 10668414 TI - Neuroprotective "agents" in surgery. Secret "agent" man, or common "agent" machine? AB - The search for clinically-effective neuroprotective agents has received enormous support in recent years--an estimated $200 million by pharmaceutical companies on clinical trials for traumatic brain injury alone. At the same time, the pathophysiology of brain injury has proved increasingly complex, rendering the likelihood of a single agent "magic bullet" even more remote. On the other hand, great progress continues with technology that makes surgery less invasive and less risky. One example is the application of endovascular techniques to treat coronary artery stenosis, where both the invasiveness of sternotomy and the significant neurological complication rate (due to microemboli showering the cerebral vasculature) can be eliminated. In this paper we review aspects of intraoperative neuroprotection both present and future. Explanations for the slow progress on pharmacologic neuroprotection during surgery are presented. Examples of technical advances that have had great impact on neuroprotection during surgery are given both from coronary artery stenosis surgery and from surgery for Parkinson's disease. To date, the progress in neuroprotection resulting from such technical advances is an order of magnitude greater than that resulting from pharmacologic agents used during surgery. The progress over the last 20 years in guidance during surgery (CT and MRI image-guidance) and in surgical access (endoscopic and endovascular techniques) will soon be complemented by advances in our ability to evaluate biological tissue intraoperatively in real-time. As an example of such technology, the NASA Smart Probe project is considered. In the long run (i.e., in 10 years or more), pharmacologic "agents" aimed at the complex pathophysiology of nervous system injury in man will be the key to true intraoperative neuroprotection. In the near term, however, it is more likely that mundane "agents" based on computers, microsensors, and microeffectors will be the major impetus to improved intraoperative neuroprotection. PMID- 10668415 TI - Brain adenosine receptors as targets for therapeutic intervention in neurodegenerative diseases. AB - Adenosine acts as a neurotransmitter in the brain through the activation of four specific G-protein-coupled receptors (the A1, A2A, A2B, and A3 receptors). The A1 receptor has long been known to mediate neuroprotection, mostly by blockade of Ca2+ influx, which results in inhibition of glutamate release and reduction of its excitatory effects at a postsynaptic level. However, the development of selective A1 receptor agonists as antiischemic agents has been hampered by their major cardiovascular side effects. More recently, apparently deleterious effects have been reported following the activation of other adenosine receptor subtypes, namely, the A2A and the A3 receptors. In particular, selective A2A receptor antagonists have been demonstrated to markedly reduce cell death associated with brain ischemia in the rat, suggesting that the cerebral A2A receptor may indeed contribute to the development of ischemic damage. The beneficial effects evoked by A2A antagonists may be due to blockade of presynaptic A2A receptors (which are stimulatory on glutamate release) and/or to inhibition of A2A receptor-mediated activation of microglial cells. Even more puzzling data have been reported for the A3 receptor subtype, which can indeed mediate both cell protection and cell death, simply depending upon the degree of receptor activation and/or specific pathophysiological conditions. In particular, a mild subthreshold activation of this receptor has been associated with a reinforcement of the cytoskeleton and reduction of spontaneous apoptosis, which may play a role in "ischemic preconditioning" of the brain, according to which a short ischemic period may protect the brain from a subsequent, sustained ischemic insult that would be lethal. In contrast, a robust and prolonged activation of the A3 receptor has been shown to trigger cell death by either necrosis or apoptosis. Such apparently opposing actions may be reconciled by hypothesizing that adenosine-mediated cell killing during ischemia may be aimed at isolating the most damaged areas to favor those parts of the brain that still retain a chance for functional recovery. In fact, both A3 receptor-mediated cell death and A2A receptor-mediated actions may be viewed as an attempt to selectively kill irreversibly damaged cells in the "core" ischemic area, in order to save space and energy for the surrounding live cells in the "pneumbra" area. Hence, the pharmacological modulation of the A2A and A3 receptors via selective ligands may represent a novel strategy in the therapeutic approach to pathologies characterized by acute or chronic neurodegenerative events. PMID- 10668416 TI - Stimulation of adenosine A3 receptors in cerebral ischemia. Neuronal death, recovery, or both? AB - The role of the adenosine A3 receptor continues to baffle, and, despite an increasing number of studies, the currently available data add to, rather than alleviate, the existing confusion. The reported effects of adenosine A3 receptor stimulation appear to depend on the pattern of drug administration (acute vs. chronic), dose, and type of the target tissue. Thus, while acute exposure to A3 receptor agonists protects against myocardial ischemia, it is severely damaging when these agents are given shortly prior to cerebral ischemia. Mast cells degranulate when their A3 receptors are stimulated. Degranulation of neutrophils is, on the other hand, impaired. While reduced production of reactive nitrogen species has been reported following activation of A3 receptors in collagen induced arthritis, the process appears to be enhanced in cerebral ischemia. Indeed, immunocytochemical studies indicate that both pre- and postischemic treatment with A3 receptor antagonist dramatically reduces nitric oxide synthase in the affected hippocampus. Even more surprisingly, low doses of A3 receptor agonists seem to enhance astrocyte proliferation, while high doses induce their apoptosis. This review concentrates on the studies of cerebral A3 receptors and, based on the available evidence, discusses the possibility of adenosine A3 receptor serving as an integral element of the endogenous cerebral neuroprotective complex consisting of adenosine and its receptors. PMID- 10668417 TI - Excitotoxicity, oxidative stress, and the neuroprotective potential of melatonin. AB - The brain consumes large quantities of oxygen relative to its contribution to total body mass. This, together with its paucity of oxidative defense mechanisms, places this organ at risk for damage mediated by reactive oxygen species. The pineal secretory product melatonin possesses broad-spectrum free radical scavenging and antioxidant activities, and prevents kainic acid-induced neuronal lesions, glutathione depletion, and reactive oxygen species-mediated apoptotic nerve cell death. Melatonin's action is thought to involve electron donation to directly detoxify free radicals such as the highly toxic hydroxyl radical, which is a probable end-product of the reaction between NO. and peroxynitrite. Moreover, melatonin limits NO.-induced lipid peroxidation, inhibits cerebellar NO. synthase, scavenges peroxynitrite, and alters the activities of enzymes that improve the total antioxidative defense capacity of the organism. Melatonin function as a free radical scavenger and antioxidant is likely facilitated by the ease with which it crosses morphophysiological barriers, e.g., the blood-brain barrier, and enters cells and subcellular compartments. Pinealectomy, which eliminates the nighttime rise in circulating and tissue melatonin levels, worsens both reactive oxygen species-mediated tissue damage and brain damage after focal cerebral ischemia and excitotoxic seizures. That melatonin protects against hippocampal neurodegeneration linked to excitatory synaptic transmission is fully consistent with the last study. Conceivably, the decreased melatonin secretion that is documented to accompany the aging process may be exaggerated in populations with dementia. PMID- 10668418 TI - Neuroprotective role of melatonin in methamphetamine- and 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine-induced dopaminergic neurotoxicity. PMID- 10668419 TI - Neuroprotective and cognitive enhancing effects of novel small peptides. PMID- 10668420 TI - Estrogens: neuroprotective or neurotoxic? AB - The present paper reviews the major modes of action of estrogen on the molecular, cellular, tissue, and neurobehavioral levels of mammalian physiology, with an emphasis on the brain as an estrogen target tissue. We draw a distinction between receptor- and nonreceptor-mediated actions, as well as delineate the range of different signal transduction pathways that might be available within a given tissue to mediate estrogenic effects. We consider species differences relevant to understanding the predictability of effects in humans from data obtained in rats or monkeys. Finally, we emphasize the importance of developmental stage in determining whether estrogenic effects are beneficial or harmful; "neuroprotective" or "neurotoxic." PMID- 10668421 TI - Role of glycemia in acute spinal cord injury. Data from a rat experimental model and clinical experience. AB - While experimental and clinical evidence indicates that in brain injury blood glucose increases with injury severity and hyperglycemia worsens neurological outcome, the role of blood glucose in secondary mechanisms of neuronal damage after acute spinal cord injury has not yet been investigated. Data from spinal cord ischemia models suggests a deleterious effect of hyperglycemia, likely due to enhanced lactic acidosis, which is primarily dependent on the amount of glucose available to be metabolized. The purpose of this study is to summarize preliminary experimental and clinical observations on the role of blood glucose in acute spinal cord injury. Between 1995 and 1996 we used the New York University (NYU) rat spinal cord injury model to test the following hypotheses: 1) Blood glucose levels increase with injury severity. 2) Fasting protects from hyperglycemia and prevents secondary damage to the spinal cord. 3) Postinjury induced hyperglycemia (dextrose 5% 2 gm/Kg) enhances spinal lesion volume. From a clinical perspective, we reviewed blood glucose records of 47 patients admitted to the Department of Neurosrgery in Verona, between 1991 and 1995, within 24 hours of acute spinal cord injury in order to determine: a) the incidence of hyperglycemia (> 140 mg/dl); b) the correlation between blood glucose and injury severity; and c) the role of methylprednisolone in affecting blood glucose. Results indicate that in a graded spinal cord injury model: 1) Early after injury, more severe contusions support significantly higher blood glucose levels. 2) Fasting overnight does not directly affect spinal cord lesion volume but influences blood gases, and we observed that a slightly systemic acidosis plays a minor neuroprotective role. Fasting also ensures more consistent normoglycemic baseline blood glucose values. 3) Postinjury-induced moderate hyperglycemia (160 190 mg/dl) does not significantly affect spinal cord injury. In the clinical study, we observed that during the first 24 hours after spinal cord injury: a) Glycemia ranges between 90 and 243 mg/dl (mean value 143 mg/dl), and close to 50% of the patients present blood glucose values higher than normal. b) Methylprednisolone administration is not associated to significantly higher blood glucose levels. c) There is a trend for larger glucose rises with more severe injury. PMID- 10668422 TI - N-acetylserotonin, melatonin and their derivatives improve cognition and protect against beta-amyloid-induced neurotoxicity. AB - After a single injection of cholinergic neurotoxin ethylcholine aziridinium (AF64A, 3 nmol intracerebroventricularly (i.c.v.)), rats failed to perform the tasks in the active avoidance (learning and retention paradigms) and water maze tests. N-Acetylserotonin (NAS), melatonin and their newly synthesized derivatives, CA-15 and CA-18, (0.3-3.0 mg/kg daily for 12-14 days) reversed the effect of AF64A in a dose-dependent manner with CA-18 being the most active. Melatonin and NAS caused sedation absent in CA-18-treated rats. The studied compounds (25-500 microM for 72 hr) protected against beta-amyloid peptide (beta AP) fragment 25-35-induced neurotoxicity in cerebellar granule cell culture. Our results suggest that neuroprotecting properties of these compounds might mediate their cognition-enhancing effects. The results obtained warrant the further search for the novel types of safe neuroprotectors among the synthetic NAS/melatonin derivatives. PMID- 10668423 TI - Neuroprotective action of bilirubin against oxidative stress in primary hippocampal cultures. PMID- 10668425 TI - Primary cultures of rat cerebellar granule cells as a model to study neuronal 5 lipoxygenase and FLAP gene expression. AB - Aging is associated with chronic neurodegenerative diseases and increased brain vulnerability that may lead to a worse outcome from brain insults in elderly than in young subjects. Inflammation is one of the patholphysiological mechanisms of both chronic and acute neurodegeneration. Leukotrienes are inflammatory lipid mediators whose formation from arachidonic acid is initiated by 5-lipoxygenase (5 LO). 5-LO is also expressed in neurons and can be activated by brain injuries, whereas 5-LO inhibitors can provide neuroprotection. The expression of the 5-LO gene appears to be inhibited by the pineal hormone, melatonin, and stimulated by stress hormone glucocorticoids (e.g., corticosterone and the synthetic glucocorticoid dexamethasone). Melatonin deficiency and hyperglucocorticoidemia frequently develop with aging. We found that old or pinealectomized, i.e., melatonin-deficient rats are more susceptible to kainate-triggered excitotoxic limbic brain injury than the corresponding young or sham-pinealectomized controls, and that pinealectomy, aging, or glucocorticoid treatment result in an enhanced expression of 5-LO in limbic structures. We hypothesize that an aging brain is at a higher risk of neurodegeneration via aging-suppressed melatonin secretion and/or aging-increased glucocorticoid secretion and the resultant upregulation of 5-LO expression. Furthermore, we propose that suppressing the 5 LO expression and/or activity will increase the brain's resistance to injury. The results of our ongoing research are expected to elucidate the role of 5-LO in aging and neurodegeneration and to indicate neuroprotective therapies that would target the 5-LO pathway. PMID- 10668424 TI - Protective effect of L-carnitine in the neurotoxicity induced by the mitochondrial inhibitor 3-nitropropionic acid (3-NPA). PMID- 10668426 TI - Intraneuronal ion distribution during experimental oxygen/glucose deprivation. Routes of ion flux as targets of neuroprotective strategies. AB - Ischemic neuronal injury appears to be mediated by disruption of subcellular ion distribution and, therefore, prevention of ion relocation might be neuroprotective. X-ray microanalysis was used to measure concentrations of Na, K, Ca and other elements in subcellular compartments (e.g., mitochondria) of CA1 neurons from oxygen/glucose-deprived (OGD) hippocampal slices. Results showed that OGD produced progressive loss of ion regulation in CA1 cells. Post-OGD reperfusion with normal media exacerbated the initial ion deregulation. To study neuroprotective mechanisms, we determined the ability of hypothermia (31 degrees C) or ion channel blockade to retard intraneuronal ion disruption induced by OGD/reperfusion. Whereas Ca2+ channel blockade (omega-conotoxin MVIIC, 3 microM) was ineffective, hypothermia and Na+ channel blockers (tetrodotoxin, TTX, 1 microM; lidocaine, 200 microM) reduced ion deregulation in subneuronal compartments. Blockade of glutamate receptors (AMPA, 10 microM; the non-NMDA receptor antagonist CNQX, 10 microM/100 microM glycine; the NMDA receptor antagonist CCP, 100 microM) during OGD/reperfusion provided nearly complete protection. These findings provide a foundation for identifying potential pharmacotherapeutic approaches and for discerning corresponding mechanisms of neuroprotection. PMID- 10668427 TI - Matrix remodeling after stroke. De novo expression of matrix proteins and integrin receptors. AB - Following an ischemic insult to the central nervous system a reorganization of cells and tissue takes place as the surrounding cells attempt to limit the injury, repair the damage, and restore normal architecture of the brain. This tissue remodeling requires de novo synthesis of genes and proteins which enables cells to actively change their relationship with the existing extracellular matrix and with other cells to reorganize the damaged tissue. We have identified two key molecular components of the matrix remodeling process after focal ischemia: osteopontin (OPN) and its integrin receptor alpha v beta 3 (alpha v beta 3). OPN is initially expressed by activated macrophages and microglia in the periinfarct region (24-48 hr) and at later times (5-15 days) in the core infarct. After focal stroke the alpha v beta 3 was upregulated by astrocytes in the periinfarct region. Spatial and temporal analyses demonstrated that at 5 days after injury the alpha v beta 3-positive astrocytes were at a distance from the osteopontin-expressing macrophages; by 15 days the alpha v beta 3-expressing astrocytes were localized within an osteopontin-rich matrix. In vitro OPN was shown to induce migration of astrocytes in a Boyden chamber system. These data suggest that OPN derived from microglia at the infarct border zone (and possible macrophages in the infarct core) may serve as an "astrokine" (suggested term for astrocyte chemoattractant) to organize the astrocyte scar after focal stroke. Our data demonstrate profound changes in brain matrix remodeling after focal ischemic stroke, including the synthesis and release of matrix proteins alien to the normal brain, the expression of integrin receptors that ligate these proteins, and possibly a novel function for microglial-derived OPN in astrocyte migration after focal ischemia that may drive glial activation, organization, and repair functions. PMID- 10668428 TI - Metallothioneins attenuate methylmercury-induced neurotoxicity in cultured astrocytes and astrocytoma cells. AB - Metallothionein-I (MT-I) was expressed in neonatal rat primary astrocyte cultures and an astrocytoma cell line by pGFAP-MT-I plasmid transfection under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Following transient transfection of the pGFAP-MT-I plasmid, MT-I mRNA and MT-I protein levels were determined by Northern blot and immunoprecipitation analyses, respectively. The ability of cells overexpressing MT-I to withstand acute methylmercury (MeHg) treatment was measured by the release of preloaded Na2(51)CrO4, an indicator of membrane integrity. Transfection with the pGFAP-MT-I plasmid led to increased mRNA (2.5-fold in astrocytes and 7.4-fold in astrocytomas) and MT-I protein (2.4-fold in astrocytes and 4.0-fold in astrocytomas) levels compared with their respective controls. Increased expression of MT-I was associated with attenuated release of Na2(51)CrO4 upon MeHg (5 microM) treatment. These results demonstrate that MT-I can be highly expressed both in primary astrocyte cultures and astrocytomas by pGFAP-MT-I plasmid transfection, and lend credence to the hypothesis that increased expression of MT-I affords protection against the cytotoxic effects of MeHg. Taken together, the data suggest that MTs offer effective cellular adaptation of MeHg cytotoxicity. PMID- 10668429 TI - Implication of poly (ADP-ribose) polymerase (PARP) in neurodegeneration and brain energy metabolism. Decreases in mouse brain NAD+ and ATP caused by MPTP are prevented by the PARP inhibitor benzamide. AB - Poly(ADP-ribose) polymerase (PARP) is a DNA binding protein that uses nicotinamide adenine dinucleotide (NAD+) as a substrate. Evidence from in vitro studies on nonneuronal cells in culture have shown that when fully activated by free radical-induced DNA damage, PARP depletes cellular NAD+ and consequently adenosine triphosphate (ATP) levels within a matter of minutes, and that this depletion is associated with a cell death that can be prevented by PARP inhibitors. The present in vivo study utilized the 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-treated mouse, a model of central nigrostriatal dopamine neurotoxicity that recapitulates certain features of Parkinson's disease (PD), and one in which we have previously shown PARP inhibitors to be protective, to examine whether MPTP acutely caused region- and time-dependent changes in levels of NAD+ and ATP in the brain in vivo and whether such effects were modified by treatments with neuroprotective doses of the PARP inhibitor benzamide. The results confirm that MPTP reduces striatal ATP levels, as previously reported by Chan et al., show that MPTP causes a regionally-selective (striatal and midbrain) loss of NAD+, and indicate that the PARP inhibitor benzamide can prevent these losses without interfering with MPTP-induced striatal dopamine release. These findings suggest an involvement of PARP in the control of brain energy metabolism during neurotoxic insult, provide further evidence in support of the participation of PARP in MPTP-induced neurotoxicity in vivo and suggest that PARP inhibitors might be beneficial in the treatment of PD. PMID- 10668430 TI - Neuroprotection against ischemia by metabolic inhibition revisited. A comparison of hypothermia, a pharmacologic cocktail and magnesium plus mexiletine. AB - Previous studies have suggested that metabolic inhibition is neuroprotective, but little evidence has been provided to support this proposal. Using the in vitro rabbit retina preparation as an established model of the central nervous system (CNS), we measured the rate of glucose utilization and lactate production, and the light-evoked compound action potentials (CAPs) as indices of neuronal energy metabolism and electrophysiologic function, respectively. We examined the effect of three (3) treatments options: hypothermia (i.e., 33 degrees C and 30 degrees C), a six-member pharmacologic "cocktail" (tetrodotoxin (0.1 microM), 2-amino-4 phosphonobutyric acid (20 microM), 2-amino-5-phosphonovaleric acid (1 mM), amiloride (1 mM), magnesium (10 mM) and lithium (10 mM) and the combination of magnesium (Mg2+ 1 mM) and mexiletine (Mex, 300 microM) on in vitro rabbit retinas, to see if there is a correlation between neuronal energy metabolism during ischemia (simulated by the reduction of oxygen from 95% to 15% and glucose from 6 mM to 1 mM), and the subsequent recovery of function. Hypothermia and the "cocktail" significantly inhibited both the rate of glucose utilization and lactate production, whereas Mg2+ and/or Mex showed only a nonsignificant tendency toward a reduction, compared to control retinas. Recovery of light-evoked CAPs was significantly improved in hypothermia- and cocktail-treated retinas, as well as with retinas exposed to the combination of Mg2+ plus Mex, but not with Mg2+ or Mex alone, relative to control retinas. A linear regression analysis of the % recovery of function versus the % reduction in the rate of glucose utilization during ischemia showed a significant correlation (r2 = 0.80, correlation coefficient = 0.9, p < 0.05) between these two parameters. This and other data discussed provide convincing evidence that there is a correlation between metabolic inhibition, achieved during ischemia, and neuroprotection. PMID- 10668431 TI - Calpeptin and methylprednisolone inhibit apoptosis in rat spinal cord injury. AB - Intracellular free Ca2+ and free radicals are increased following spinal cord injury (SCI). These can activate calpain to degrade cytoskeletal proteins leading to apoptotic and necrotic cell death. Primary injury triggers a cascade of secondary injury, which spreads to rostral and caudal areas. We tested calpain involvement in apoptosis in five 1-cm segments of rat spinal cord with injury (40 g-cm) induced at T12 by weight-drop. Animals were immediately treated with calpeptin (250 micrograms/kg) and methylprednisolone (165 mg/kg) and sacrificed at 48 hr. Untreated SCI rats manifested 68-kD neurofilament protein (NFP) degradation (indicating calpain activity), and internucleosomal DNA fragmentation (indicating apoptosis). Both calpain activity and apoptosis were highest in the lesion, and decreased with increasing distance from the lesion. Treatment decreased 68-kD NFP degradation with reduction in apoptosis in all five areas. Thus, calpeptin and methylprednisolone are found to be neuroprotective in SCI. PMID- 10668432 TI - Calpastatin is upregulated and acts as a suicide substrate to calpains in neonatal rat hypoxia-ischemia. PMID- 10668433 TI - High extracellular glutamate and neuronal death in neurological disorders. Cause, contribution or consequence? AB - In models of neurological disorders, increased extracellular glutamate and beneficial effects produced by glutamate-receptor antagonists are consistently taken as supporting evidence of excitotoxicity. This systematic interpretation is over-simplified and potentially misleading. High extracellular glutamate is not a reliable indicator of endogenous excitotoxicity, i.e., the intrinsic, potential neurotoxicity of endogenous glutamate whenever it accumulates extracellularly. Firstly, because the extracellular levels of glutamate necessary to produce depolarization and death in vivo, are far above those measured in models of neurological disorders. Secondly, because changes in the concentration of glutamate in the synaptic cleft (i.e., the relevant compartment for endogenous excitotoxicity) are not reflected extracellularly. Protection by glutamate receptor antagonists does not necessarily imply inhibition of excitotoxic abnormalities. Indeed, neuronal death initiated by insults such as ischemia results from multifactorial processes that may be interrelated. Therefore, beneficial effects resulting from an interaction with glutamate-mediated transmission may actually render the cell more resistant to other deleterious mechanisms (e.g., mitochondrial injury, oxidative stress). PMID- 10668434 TI - Evidence disputing the importance of excitotoxicity in hippocampal neuron death after experimental traumatic brain injury. AB - The hippocampus is selectively vulnerable to experimental traumatic brain injury (TBI). Beneficial effects of glutamate receptor antagonists and increased extracellular levels of glutamate have suggested that glutamate-mediated excitotoxicity may be responsible for this selective damage. In order to clarify this important issue, we applied a severe parasagittal fluid percussion injury (FPI) to strains of mice shown to be susceptible and resistant to kainic acid (KA)-induced excitotoxic hippocampal damage. Dystrophic neurons were present by 10 min after FPI in the hippocampi of both strains. Damaged hippocampal neurons were absent at 4 days and 7 days. Additionally, there was no significant difference (p = 1.00) in CA3 neuron survival between KA-susceptible and resistant mice at 4 days. In conclusion, excitotoxicity does not significantly contribute to hippocampal neuron loss after FPI and, in contrast to classic studies of excitotoxicity in vivo, the pattern of hippocampal cell death after TBI is extremely acute. PMID- 10668435 TI - Neuroprotective properties of nitric oxide. AB - The discoveries of physiological roles of nitric oxide (.NO) as the mediator of endothelium-derived relaxing factor (EDRF) action and the activator of guanylyl cyclase to increase cyclic guanosine monophosphate (cGMP), which lead to vasorelaxation in the cardiovascular system, have been awarded with the 1998 Nobel Prize of Medicine. The present review discusses putative beneficial effects of .NO in the central nervous system (CNS). In addition to its prominent roles of the regulation of cerebral blood flow and the modulation of cell to cell communication in the brain, recent in vitro and in vivo results indicated that .NO is a potent antioxidative agent. .NO terminates oxidant stress in the brain by (i) suppressing iron-induced generation of hydroxyl radicals (.OH) via the Fenton reaction, (ii) interrupting the chain reaction of lipid peroxidation, (iii) augmenting the antioxidative potency of reduced glutathione (GSH) and (iv) inhibiting cysteine proteases. It is apparent that .NO--a relative long half-life nitrogen-centered weak radical--scavenges those short-lived, highly reactive free radicals such as superoxide anion (O2.-), .OH, peroxyl lipid radicals (LOO.) and thiyl radicals (i.e., GS.), yielding reactive nitrogen species including nitrites, nitrates, S-nitrosoglutathione (GSNO) and peroxynitrite (ONOO-). GSNO is 100-fold more potent than GSH; it completely inhibits the weak peroxidative effect of ONOO-. Moreover, CO2 and .NO neutralize prooxidative effects of ONOO-. CO2 prevents protein oxidation but not 3-nitrotyrosine formation caused by ONOO-. Finally, neuroprotective effects of GSNO and .NO have been demonstrated in brain preparations in vivo. These novel neuroprotective properties of .NO and GSNO may have their physiological significance, since oxidative stress depletes GSH while increasing GS. and .NO formation in astroglial and endothelial cells, resulting in the generation of a more potent antioxidant GSNO and providing additional neuro-protection at microM concentrations. This putative GSNO pathway (GSH-->GS.- >GSNO-->.NO + GSSG-->GSH) may be an important part of endogenous antioxidative defense system, which could protect neurons and other brain cells against oxidative stress caused by oxidants, iron complexes, proteases and cytokines. In conclusion, .NO is a potent antioxidant against oxidative damage caused by reactive oxygen species, which are generated by Fenton reaction or other mechanisms in the brain via redox cycling of iron complexes. PMID- 10668436 TI - Neuroprotective strategies for HIV-1-associated neurologic disease. PMID- 10668437 TI - Changes in mRNA levels for heat-shock/stress proteins (Hsp) and a secretory vesicle associated cysteine-string protein (Csp1) after amphetamine (AMPH) exposure. AB - Damage to nerve terminals, reactive gliosis and somatic degeneration can result when pronounced hyperthermia occurs during amphetamine (AMPH) exposure. The effects of AMPH-induced hyperthermia and damage on the relative mRNA levels for several heat shock/stress proteins (Hsp27, Hsp60, Hsp70 and Hsc70), as well as secretory vesicle associated cysteinestring protein (Csp1) were determined in both the striatum and substantia nigra using reverse transcriptase polymerase chain reaction (RT-PCR). These changes were compared to changes in Hsp mRNA levels seen in primary rat cerebral astrocyte cultures after heat shock/stress. Striatal Hsp70 mRNA increased about 2-fold over control levels at 16 hr after AMPH-induced hyperthermia, and was the only Hsp species to significantly increase in response to AMPH. Hsp70 mRNA levels returned to control within 14 days after AMPH. Two-fold increases in Hsp70 mRNA were also seen in primary cultures of rat cerebrum 24 hr after heat shock. In primary cultures and brain tissue, the increased Hsp70 mRNA levels were still more than 500-fold less than constitutive Hsc70 mRNA and 50-fold less than Hsp60 levels. Hsp27 mRNA was not present in the striatum, nigra and primary cell cultures. Thus, the expression of Hsp species mRNA measured was very similar in brain tissue and primary cell cultures. Because only a modest induction of Hsp 70 mRNA occurred, the Hsp species evaluated may only play a minor role in AMPH neurotoxicity. However, further studies are necessary to determine whether large increases in Hsp70 are occurring in selected neurons or glia in the striatum. RT-PCR products for Csp1 were produced in total RNA obtained from brain but not from cultured astrocytes, suggesting that the Csp1 mRNA measured by RT-PCR is of neuronal origin. Csp1 mRNA levels were acutely downregulated in neurons in the substantia nigra, possibly in response to damage, but not the striatum after AMPH exposure. A slight long-term upregulation at 4 months of Csp1 mRNA may occur in the striatum but not in nigra. PMID- 10668438 TI - Benzamide, a poly(ADP-ribose) polymerase inhibitor, is neuroprotective against soman-induced seizure-related brain damage. PMID- 10668439 TI - Central noradrenergic neurotoxicity of DSP4 in mice. Studies on the neuroprotective potential of the poly(ADP-ribose) polymerase inhibitor, benzamide. PMID- 10668440 TI - The antioxidative effect of carboxyfullerenes (C3/D3) on iron-induced oxidative injury in CNS. AB - Carboxyfullerenes, including two regioisomers C3 and D3, were investigated as antioxidants against iron-induced oxidative stress in vivo and in vitro. Both C3 and D3 dose-dependently inhibited autoxidation and iron-elevated lipid peroxidation in cortical homogenates. The antioxidative property of C3 was compared to Trolox (a water-soluble analogue of vitamin E) and glutathione. C3 was more effective than glutathione but was less effective than Trolox in inhibiting iron-induced elevation in lipid peroxidation. In urethane-anesthetized rats, intranigral infusion of iron degenerated the nigrostriatal dopaminergic system, including as elevation in lipid peroxidation in the infused substantia nigra (SN) and reductions in K(+)-evoked dopamine overflow and dopamine content in the ipsilateral striatum 7 days after the infusion. Local application of iron with C3 or D3 prevented iron-induced oxidative injuries. Our data suggest that carboxyfullerenes have a neuroprotective effect in preventing iron-induced oxidative injury in CNS. PMID- 10668441 TI - Neuregulin signaling in brain injury and in animal models of ischemia. PMID- 10668442 TI - Protective and rescuing abilities of IGF-I and some putative free radical scavengers against beta-amyloid-inducing toxicity in neurons. AB - beta-Amyloid (A beta) peptides are most likely involved in the neurodegenerative process occurring in Alzheimer's Disease (AD) and are enriched in senile plaques. The mechanisms of A beta toxicity are not clear but likely involve free radicals and apoptosis. Much interest is currently aiming at developing effective approaches to block A beta toxicity in order to slow down disease progression. In that context, we are particularly interested in studying the role of insulin-like growth factors, particularly IGF-I and purported free radical scavengers including a Gingko biloba extract (EGb761) as blocker of A beta toxicity in a simple in vitro model of hippocampal primary cultures. We observed that both IGF I and EGb761 are unique in that they are able not only to protect but even to rescue neurons against A beta toxicity. These results are summarized here and possible mechanisms of action are discussed to explain the protective properties of these two classes of agents. PMID- 10668443 TI - The unique histopathological responses of the injured spinal cord. Implications for neuroprotective therapy. AB - Tissue destruction at the primary site of a spinal cord injury leads to persistent necrosis that progressively enlarges the lesion. Steroids attenuate this necrotizing process and promote tissue repair even though such anti inflammatory drugs interfere with wound healing in non-CNS organs. To address this paradox, the spinal cord of rats and mice was crushed extradurally and the effects of the following anti-inflammatory agents studied by light microscopical image analysis: allopurinol, aminoguanidine, indomethacin, a bacterial lipopolysaccharide, naproxen, and pregnenolone. The contribution of Wallerian degeneration to progressive necrosis was studied in a mutant mouse strain (WldS) that is characterized by delayed Wallerian degeneration. In rats, the anti inflammatory agents selectively attenuated progressive necrosis and encouraged wound healing. In mice, considerable tissue repair occurred without pharmacological intervention; this wound-healing process was delayed in the mutant WldS strain. Since spinal cord injury results in concomitant tissue necrosis and wound healing, a goal of neuroprotective therapy is to regulate the dynamic balance between these destructive and reparative processes. PMID- 10668444 TI - Ion channel modulation as the basis for neuroprotective action of MS-153. AB - MS-153, (R)-(-)-5-methyl-1-nicotinoyl-2-pyrazoline, is a new neuroprotective drug. Recent data in the literature suggest that it inhibits glutamate accumulation occurring during ischemia and the translocation of protein kinase C gamma (PKC gamma). The present study was undertaken to prove the hypothesis that MS-153 blocks neuroreceptors and ion channels involved in glutamate accumulation. Neurons isolated from rat dorsal root ganglia and frontal cortex were used for recording channel currents by the whole-cell patch clamp technique. The effects of bath-applied MS-153 were examined on tetrodotoxin-sensitive and tetrodotoxin resistant sodium channels and high voltage-gated calcium channels of dorsal root ganglion neurons, and channels activated by glutamate, N-methyl-D-aspartate (NMDA), kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxarole propionic acid (AMPA), gamma-aminobutyric acid (GABA) and acetylcholine (ACh) in cortical neurons. MS-153 at a concentration of 300 microM had no effect on either tetrodotoxin-sensitive or tetrodotoxin-resistant sodium channels. High voltage gated calcium channels were either suppressed or not affected by 1-300 microM MS 153. The variable blocking effect of MS-153 was due to the variable activity of intracellular components in individual neurons, especially that of PKC, whose translocation is known to be inhibited by MS-153. When 100 nM phorbol 12 myristate-13-acetate (PMA) was applied to neurons, MS-153 suppressed the calcium channel current more frequently. Calphostin C (0.5 microM), a specific PKC inhibitor, applied intracellularly via recording patch pipette, completely abolished MS-153 suppression of the calcium channel current. Currents induced by glutamate, NMDA, kainate, AMPA, GABA or ACh were not affected by MS-153 at 300 microM. It was concluded that MS-153 inhibited high voltage-gated calcium channels through interactions with PKC, thereby preventing massive release of glutamate from nerve terminals in ischemic conditions. PMID- 10668445 TI - Blockade of NAALADase: a novel neuroprotective strategy based on limiting glutamate and elevating NAAG. AB - Excessive glutamate receptor activation is thought to be involved in the neuronal injury caused by stroke. Based on the hypothesis that N-acetyl-aspartyl-glutamate (NAAG) is a modulatory neurotransmitter or storage form of glutamate, we have pursued a novel strategy of therapeutic intervention, blockade of N-acetylated alpha-linked acidic dipeptidase (NAALADase), the enzyme that hydrolyzes NAAG to liberate glutamate. Using the suture model of transient middle cerebral artery occlusion (MCAO) in rats, the prototype NAALADase inhibitor 2 (phosphonomethyl)pentanedioic acid (2-PMPA) dramatically reduced extracellular glutamate accumulation measured by microdialysis both during a 2-hour occlusion and during reperfusion, consistent with an effect on glutamate supply. During reperfusion, the decrease in glutamate was accompanied by an equimolar, reciprocal rise in extracellular NAAG. NAALADase inhibition may prove to be a well tolerated therapy for cerebral ischemia. In addition, NAALADase inhibitors should prove to be important tools in understanding the physiological role of NAAG in the brain. PMID- 10668446 TI - The low-affinity, use-dependent NMDA receptor antagonist AR-R 15896AR. An update of progress in stroke. AB - Use-dependent N-methyl-D-aspartate (NMDA) receptor antagonists protect neurons from the lethal consequences of excessive stimulation by excitatory amino acids. Clinical development of high-affinity compounds such as MK801 have been limited due to untoward side effects. Toward this end, the lower-affinity use-dependent NMDA antagonists have greater margins of safety and have advanced to clinical trials for stroke, epilepsy, head trauma and chronic neurodegenerative disorders. AR-R 15896AR is currently in Phase II trials for stroke and has been repeatedly demonstrated to afford neuroprotection in a variety of in vivo and in vitro models associated with ischemia/excitotoxic conditions. PMID- 10668447 TI - Intracellular survival pathways against glutamate receptor agonist excitotoxicity in cultured neurons. Intracellular calcium responses. AB - Cultured rat cerebellar granule cells are resistant to the excitotoxic effects of N-methyl-D-aspartate (NMDA) and non-NMDA receptor agonists under three conditions: 1) prior to day seven in vitro when cultured in depolarizing concentrations of potassium [25 mM]; 2) at any time in vitro when cultured in non depolarizing concentrations of potassium 5 mM[; and 3) when neurons, cultured in depolarizing concentrations of potassium 25 mM[ for eight days in vitro, are pretreated with a subtoxic concentration of NMDA. The focus of this paper is to determine: a) whether the resistance to excitotoxicity by NMDA and non-NMDA receptor agonists is due to a decreased intracellular calcium Ca++[i response to glutamate receptor agonists in cultured rat cerebellar granule cells; or b) whether Ca++[i levels induced by the agonists are similar to those observed under excitotoxic conditions. Granule cells, matured in non-depolarizing growth medium, treated with glutamate resulted in an increase in Ca++[i followed by a plateau that remained above baseline in virtually all neurons that responded to glutamate. The response was rapid in onset (< 10 sec) and the pattern of response heterogeneous in that cells responsive to glutamate increased their Ca++[i to different extents; some cells did not respond to glutamate. Kainate also produced significant elevations in Ca++[i. The Ca++[i response to glutamate in neurons matured in depolarizing (25 mM K+) growth medium for three days was rapid, transient and heterogeneous, which reached a plateau that was elevated above baseline levels; removing the glutamate markedly reduced the Ca++[i concentration. Activation of the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)/kainate receptors by kainic acid produced similar changes in Ca++[i responses. At a time when cultured cerebellar granule cells become susceptible to the excitotoxic effects of glutamate acting at NMDA receptors (day in vitro (DIV) 8) in depolarizing growth medium, glutamate elicited Ca++[i responses similar to those observed at a culture time when the neurons are not susceptible to the excitotoxic effects of glutamate (DIV 3). Pretreatment of the cultured neurons with a subtoxic concentration of NMDA, which protects all neurons against the excitotoxic effects of glutamate, did not alter the maximal Ca++[i elicited by an excitotoxic concentration of glutamate. PMID- 10668449 TI - NPS 1506, a novel NMDA receptor antagonist and neuroprotectant. Review of preclinical and clinical studies. AB - NPS 1506 is a moderate affinity, uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist. NPS 1506 is neuroprotective in rodent models of ischemic stroke, hemorrhagic stroke, and head trauma, with a 2-hr window of opportunity. Neuroprotectant doses of NPS 1506 ranged from approximately 0.1-1.0 mg/kg, with peak plasma concentrations ranging from 8-80 ng/mL. Even at doses producing behavioral toxicity, NPS 1506 did not elicit MK-801-like behaviors, did not generalize to phencyclidine (PCP), and did not elicit neuronal vacuolization. In a Phase I study, intravenous (i.v.) doses of NPS 1506 from 5-100 mg were well tolerated and provided plasma concentrations in excess of those required for neuroprotection in rodents. Adverse events at the 100-mg dose included mild dizziness and lightheadedness, and mild to moderate ataxia. Neither PCP-like psychotomimetic effects nor cardiovascular effects were noted. The long plasma half-life of NPS 1506 (approximately 60 hr) suggests that a single i.v. dose will provide prolonged neuroprotection in humans. PMID- 10668448 TI - Neuroprotective actions of novel and potent ligands of group I and group II metabotropic glutamate receptors. AB - The role of group I metabotropic glutamate (mGlu) receptors in neurodegeneration is controversial because of the contradictory effects of mGlu1/5 agonists in in vitro models of neuronal cell death. In this study, novel and selective antagonists of mGlu1 and mGlu5: LY367385 and LY367366 were found to show consistent neuroprotective effects against N-methyl-D-aspartate (NMDA)-induced excitotoxicity in vitro and in vivo. Furthermore, intraventricular administration of LY367385 reduced hippocampal cell death in gerbils subjected to transient global ischemia. Previous studies have also shown that activation of group II mGlu receptors may contribute to neuroprotective mechanisms in vitro and in vivo. Three potent group II mGlu agonists--LY354740, LY379268 and LY389795--were found to attenuate both NMDA excitotoxicity and staurosporine-induced neuronal cell death. LY354740 and LY379268 were protective against transient global ischemia in gerbils when dosed intraperitoneally. These results support the view that antagonists of mGlu1 and mGlu5 and agonists of group II mGlu receptors may be useful agents in the therapeutic treatment of neurodegenerative disease. PMID- 10668450 TI - Peroxynitrite scavengers for the acute treatment of traumatic brain injury. AB - Recent evidence has suggested that the superoxide and nitric oxide-derived reactive oxygen species peroxynitrite (ONOO-) may play a significant role in the acute pathophysiology of brain injury. One pharmacological mechanism by which ONOO(-)-mediated damage might be interrupted is by the administration of scavenging compounds such as the thiol-containing compound penicillamine. In the present study, we examined the ability of either penicillamine (Pen) or the more brain penetrable penicillamine methyl ester (PenME) (0.01, 0.1, 1.0 or 10.0 mg/kg i.v. 5 min post-injury) to improve the early (1 hr) neurological recovery (grip score) of male CF-1 mice after a severe (900 g-cm; 50 g x 18 cm) injury. Pen produced a dose-related improvement in grip score. At 1.0 mg/kg, a +112% improvement was observed compared to vehicle-treated mice, and at 10.0 mg/kg, the increase was +168% (both, p < 0.05). PenME more potently improved the 1-hr grip score, but the magnitude of the optimal effect (+96% at 0.1 mg/kg; p < 0.02) was no greater than that observed with Pen, which largely remains in the cerebral microvasculature. These results are consistent with a role of ONOO- in acute head injury, but suggest that microvascular scavenging may be of primary therapeutic importance during the early post-traumatic period. PMID- 10668451 TI - Regional cytokine, cytokine receptor and neuropeptide mRNA changes associated with behavioral and neuroanatomical abnormalities in persistent, noninflammatory virus infection of neonatal rats. PMID- 10668452 TI - Development of a nonhuman primate model for studying the consequences of long term neuroprotectant administration on complex brain functions in developing animals. PMID- 10668453 TI - Melatonin as a pharmacological agent against neuronal loss in experimental models of Huntington's disease, Alzheimer's disease and parkinsonism. AB - This review summarizes the experimental findings related to the neuroprotective role of melatonin. In particular, it focuses on research directed at models of Huntington's disease, Alzheimer's disease and Parkinsonism. Melatonin has been shown to be highly effective in reducing oxidative damage in the central nervous system; this efficacy derives from its ability to directly scavenge a number of free radicals and to function as an indirect antioxidant. In particular, melatonin detoxifies the highly toxic hydroxyl radical as well as the peroxyl radical, peroxynitrite anion, nitric oxide, and singlet oxygen, all of which can damage macromolecules in brain cells. Additionally, melatonin stimulates a variety of antioxidative enzymes including superoxide dismutase, glutathione peroxidase and glutathione reductase. One additional advantage melatonin has in reducing oxidative damage in the central nervous system is the ease with which to crosses the blood-brain barrier. This combination of actions makes melatonin a highly effective pharmacological agent against free radical damage. The role of physiological levels of melatonin in forestalling oxidative damage in the brain is currently being tested. PMID- 10668454 TI - The regulation of cerebral blood flow during intravenous cocaine administration in cocaine abusers. AB - Cocaine abuse is associated with heightened risk of life-threatening neurological complications such as strokes, seizures, and transient ischemic attacks. We used transcranial Doppler (TCD) sonography, a continuous measure of cerebral blood flow velocity, to better understand the changes in cerebral hemodynamics produced by cocaine administration, which may lead to an increased risk for stroke in cocaine abusers. Heart rate and blood pressure were also measured. Blood flow velocity of seven cocaine abusers was studied during placebo, 10-, 25-, and 50-mg intravenous (i.v.) injections of cocaine. A significant increase in mean and systolic velocity which lasted for about two minutes was observed with all doses of cocaine, with no change in the placebo condition. This increase in systolic velocity indicates that cocaine produces an immediate and brief period of vasoconstriction in large arteries of the brain. The present results elucidate the time course of cocaine's acute cerebrovascular effects and provide a better understanding of etiology of cocaine-related stroke and transient ischemic attacks. PMID- 10668455 TI - Time course of brain temperature and caudate/putamen microdialysate levels of amphetamine and dopamine in rats after multiple doses of d-amphetamine. AB - Brain temperature monitoring and microdialysis were performed simultaneously in the caudate/putamen (CPu) of conscious, freely moving rats dosed with d amphetamine (AMPH). The brain temperature was determined via a thermistor inserted through a microdialysis guide cannula located in the left CPu, while the microdialysis probe was positioned in the right CPu. The peak AMPH and dopamine (DA) levels were reached 40 to 60 min after dosing, while peak brain temperature was not achieved until 20 to 40 min thereafter in rats becoming moderately hyperthermic. Those rats becoming severely hyperthermic (temperatures above 41.0 degrees C) had microdialysate concentrations of AMPH and DA almost 2-fold higher than those with moderate hyperthermia after the second dose of 5 mg/kg AMPH. However, these peaks were not reached until 60 to 80 min after dosing. This was probably due, in part, to the longer half-life of AMPH in the severely hyperthermic group. The changes in brain temperature observed after exposure to neurotoxic doses of AMPH closely paralleled core body temperature changes previously reported during AMPH exposure. Temperature plays an important role in many types of neurotoxicity, and monitoring brain temperature during microdialysis studies can be done continuously, and with less chance of damage to the microdialysis equipment than most of the traditional methods used to measure core body temperature. PMID- 10668456 TI - Neuroprotective effects of HU-211 on brain damage resulting from soman-induced seizures. AB - Neuroprotective effects of HU-211 (dexanabinol), a synthetic nonpsychotropic analog of tetrahydrocannabinol, on brain damage resulting from soman-induced seizures were examined in male Sprague-Dawley rats challenged with 1.6 LD50 soman. At 5 or 40 min after onset of seizures, the rats were given an intraperitoneal injection of 25 mg/kg HU-211. All rats that received soman showed electrocorticographic (ECoG) evidence of sustained seizures and status epilepticus for 4-6 hr. HU-211 had no effect on either the strength or duration of seizure activity. Administration of HU-211 at 5 min after seizure onset reduced median lesion volume 86% (as assessed by microtubule-associated protein 2 (MAP2)-negative staining), and when administered 40 min post-onset, the reduction in necrosis was 81.5% despite the presence of continuous seizures for 4-5 hr. These observations were corroborated by hemotoxylin and eosin (H&E) histopathological assessment that showed a significant reduction in piriform cortical neuronal damage in HU-211-treated animals. It is concluded that HU-211 provides considerable neuroprotection against brain damage produced by soman induced seizures. PMID- 10668457 TI - Abuse liability assessment of neuroprotectants. AB - There has been considerable interest in the potential of N-methyl-D-aspartate (NMDA) receptor antagonists in the treatment of a diverse group of neurological disorders including cerebral ischemia and neurodegeneration. The amino acids L glutamate and L-aspartate have been shown to possibly mediate excitatory synaptic transmission in the central nervous system (CNS) via selective excitatory amino acid receptors. Competitive and noncompetitive antagonists acting at the NMDA receptors have been shown to possess relevant activity. However, NMDA antagonists can produce a variety of adverse neurobehavioral effects in both animals and humans. These adverse events are particularly pronounced with NMDA antagonists (phencyclidine (PCP), ketamine, and MK-801) that have dissociative anesthetic properties and block NMDA receptor-mediated responses by binding to the cation channel of the NMDA receptor complex. When a new pharmaceutical product demonstrates structural similarity and/or a similar pharmacological profile with a known drug of abuse, the characterization of its abuse potential is needed by the FDA for scientific review. The abuse liability assessment is based upon an evaluation of data on the chemistry, pharmacology (preclinical and clinical), pharmacokinetics, and pharmacodynamic profiles of the drug, and the adverse events/effects reported in clinical trials. The evaluation of the drug's abuse potential is determined relative to pharmacologically similar drugs. This includes determination of the drug's receptor binding efficacy, preclinical pharmacology, reinforcing efficacy, discriminative stimulus effects, dependence producing potential, pharmacokinetics, and assessment of the clinical efficacy safety database relative to abuse and clinical abuse liability studies. It has been well established that high-affinity noncompetitive NMDA antagonists have reinforcing efficacy and can serve as discriminative stimuli in operant procedures. In a variety of species in drug discrimination studies, each antagonist is capable of generalizing to the others, and it is believed that these effects may be mediated through the NMDA blockade. The generalization of each substance for another suggests production of common subjective effects in humans. PMID- 10668458 TI - The future of neuroprotection. PMID- 10668459 TI - Apoptosis: a two-edged sword in aging. AB - Here we summarize briefly what is known about both the positive and negative impacts of apoptosis during aging in mammalian systems and also update an earlier review. It is important to understand both of these impacts to devise useful interventions. Such interventions include both physiological and molecular approaches, including transgenic interventions. The critical roles of the mitochondria in both generating reactive oxygen species, and in initiating apoptosis are recognized, suggesting that maintaining mitochondrial function could be an important therapeutic goal, especially in post-mitotic tissues. In contrast, the ability to eliminate unwanted, damaged and dysfunctional cells through apoptosis has anti-aging implications in mitotic tissues. PMID- 10668460 TI - Ultrastructural alterations of mitochondria in pre-apoptotic and apoptotic hepatocytes of TNF alpha-treated galactosamine-sensitized mice. AB - The electron microscopical studies presented here show that characteristic morphological alterations in mitochondria are a very early hallmark of the hepatocellular apoptotic program. Before chromatin condensation occurs, the outer mitochondrial membrane is focally disrupted and the inner membrane protrudes through this gap forming a hernia. The demonstration of cytochrome oxidase in mitochondria revealed a very strong activity in pre-apoptotic and apoptotic cells as well as in apoptotic bodies. PMID- 10668461 TI - Mitochondrial membrane permeabilization during the apoptotic process. AB - Apoptosis may be viewed as a triphasic process. During the pre-mitochondrial initiation phase, very different pro-apoptotic signal transduction or damage pathways can be activated. These pathways then converge on the mitochondrion, where they cause the permeabilization of the inner and/or outer membranes with consequent release of soluble intermembrane proteins into the cytosol. The process of mitochondrial membrane permeabilization would constitute the decision/effector phase of the apoptotic process. During the post-mitochondrial degradation phase downstream caspases and nucleases are activated and the cell acquires an apoptotic morphology. Recently, a number of different second messengers (calcium, ceramide derivatives, nitric oxide, reactive oxygen species) and pro-apoptotic proteins (Bax, Bak, Bid, and caspases) have been found to directly compromise the barrier function of mitochondrial membranes, when added to isolated mitochondria. The effects of several among these agents are mediated at least in part via the permeability transition pore complex (PTPC), a composite channel in which members of the Bcl-2 family interact with sessile transmembrane proteins such as the adenine nucleotide translocator. These findings suggest that the PTPC may constitute a pharmacological target for chemotherapy and cytoprotection. PMID- 10668462 TI - Coenzyme Q10 can in some circumstances block apoptosis, and this effect is mediated through mitochondria. AB - The mitochondrial component coenzyme Q10 (CoQ10) has been used for many years as a dietary supplement intended to promote good health by trapping free radicals, thus preventing lipid peroxidation and DNA damage. We have tested its use as a generic anti-apoptotic compound and have found that its ability to protect against apoptosis varies depending on both cell type and mode of cell death induction. We have further established that this protection may be mediated by its effect on mitochondrial function and viability. We provide additional evidence that CoQ10's protective effect on mitochondrial membrane potential does not always result in altered mitochondrial enzyme activity and neither does it guarantee survival. These observations open the way for further investigations into the mechanisms involved in mitochondrial control of apoptosis. PMID- 10668463 TI - Neither caspase-3 nor DNA fragmentation factor is required for high molecular weight DNA degradation in apoptosis. AB - In this paper, we show that there is a two-step process of DNA fragmentation in apoptosis; DNA is first cleaved to large fragments of 50-300 kb that are subsequently cleaved to smaller oligonucleosomes in some, but not all cells. Significantly, only the first stage is considered essential for cell death since some cells, for example human MCF7 breast carcinoma cells and human NT2 neuronal cells, do not show this behavior but still display normal nuclear morphological apoptotic changes. In cells that usually produce small fragments blocking the second (internucleosomal) stage of DNA fragmentation prevents neither nuclear condensation nor apoptosis. We are beginning to understand why the extent of DNA fragmentation during apoptosis varies enormously and why it appears to be a function of the cell type not the inducer. Presumably, this reflects the content of not only endonuclease activit(ies) but also on the ability of the cells to activate caspases, particularly caspase-3, and other proteases that may be involved in endonuclease activation. Since NT2 cells activate caspase-3, but do not correctly process DFF45, other factors must also impinge on the inevitability of that process. PMID- 10668464 TI - Apoptosis-related functional features of the DNaseI-like family of nucleases. AB - Rat DNaseYb and its human homolog DHP2 are members of a new family of DNaseI-like endonucleases. They contain all the conserved amino acid residues to engage a DNaseI-like catalytic activity and the same molecular mechanisms of DNA hydrolysis. The sequence similarity can be extended to other families of nucleases, such as FEN-1, DNA polymerases, RNaseH and exonuclease III, involved in the ion-dependent hydrolysis of nucleic acids. Their unique features include the NLS signals that place them in the nuclei and a high content of positively charged amino acid residues that results in their high affinity for the substrate. Their properties are consistent with a role in the early stage DNA degradation during apoptosis. The caspase-DFF45/CIDE-CPAN pathway is most likely involved in the second stage of internucleosomal DNA degradation. However, cells express constitutively multiple transcripts encoding DNA degrading enzymes and related molecules, hence they have the molecular diversity to engage the self destructive pathway appropriate to a given trigger. PMID- 10668465 TI - Does the oxidative/glycolytic ratio determine proliferation or death in immune recognition? AB - Here we discuss the possibility that the way cells utilize fuel(s) for energy confers the properties that can be recognized by the immune system and, reciprocally, that recognition by the immune system can alter the balance of the cell's energy metabolism. We propose that immune recognition, of somatic cells via MHC can alter the their energy metabolism and induce a metabolic shift. We demonstrate the reciprocal relationship that inducing a shift in metabolism toward glycolysis by supplying glucose and insulin results in the upregulation of immunologically recognizable molecules such as cell surface Fas. Thus, immune recognition can induce metabolic deviation. Metabolic deviation can result in altered immune recognition and ultimately in cell proliferation, cell differentiation, or cell death. PMID- 10668466 TI - Regulation of transglutaminases by nitric oxide. AB - Nitric oxide (NO) is an inorganic diffusible molecular messenger that plays several central roles in pathophysiology. NO can affect the biological activity of proteins through the direct or indirect (via intermediate S-nitrosothiols) S nitrosylation of protein thiol groups. Transglutaminases (TGases), Ca(++) dependent enzymes that modify proteins by cross-linking reactions, require a cysteine residue in the active site as a prerequisite for their catalytic activity. Therefore, NO may regulate enzymatic activity of TGases and their biological effects, via S-nitrosylation of their crucial thiol groups. We here review the effects of NO on coagulation factor XIII (fXIII, or plasma TGase) and TGase 2 (or tissue transglutaminase). NO has an inhibitory effect on fXIII, thus suppressing the gamma-chain cross-linking in fibrin gels, and subsequent clot formation. Tissue transglutaminase, an apoptotic effector molecule, also represents a molecular target for NO. Accordingly, the inhibition of tissue transglutaminase enzymatic activity by NO is able to prevent the induction of apoptosis. PMID- 10668467 TI - The molecular mechanism of programmed cell death in C. elegans. AB - Programmed cell death or apoptosis plays a fundamental role during animal development, metamorphosis, and tissue homeostasis. It is a genetically controlled physiological process that comprises two distinct and sequential processes: the death of cells, and their subsequent removal by engulfing cells. In the nematode C. elegans, genetic studies led to the discovery of 15 genes that function in programmed cell death (FIG. 1). These 15 genes have been divided into four groups based on the order of their activity during the process of programmed cell death: (1) those involved in the decision making (ces-1 and ces-2); (2) in the process of execution (ced-3, ced-4, ced-9 and egl-1); (3) in the engulfment of dying cells by engulfing cells (ced-1, ced-2, ced-5, ced-6, ced-7, ced-10, ced 12); and (4) those in the degradation of cell corpses within engulfing cells (nuc 1). In the last five years, several genes in the genetic pathway of programmed cell death have been shown to be conserved across a wide range of species; all genes involved in the step of execution in C. elegans have their corresponding mammalian homologs (FIG. 2). Furthermore, emerging evidence from molecular studies of engulfment genes in several species suggests that the signaling process from apoptotic cells to engulfing cells and the subsequent engulfment process might be also conserved across species (TABLE 1). PMID- 10668468 TI - Developmental regulation of induced and programmed cell death in Xenopus embryos. AB - We have analyzed the role of cell death during early Xenopus development and have identified two distinct types of cell death programs during the period between fertilization and the tadpole stage. One is a maternal cell death program that is activated at the onset of gastrulation following damage to the pre-midblastula transition embryo, resulting in the death of non-viable cells. The activation of this cell death program at a specific time during development is a maternally programmed event under the control of a developmental timer set at fertilization, and does not depend on the type of stress applied, on cell cycle progression, or de novo protein synthesis. Subsequently, a second program corresponding to programmed cell death is initiated as part of the normal development of the embryo. Programmed cell death starts at the onset of gastrulation and we have analyzed its spatio-temporal patterns by a whole-mount in situ DNA end labeling technique (the TUNEL protocol). PMID- 10668469 TI - Bone morphogenetic proteins regulate interdigital cell death in the avian embryo. AB - The embryonic limb bud provides an excellent model for analyzing the mechanisms that regulate programmed cell death during development. At the time of digit formation in the developing autopod, the undifferentiated distal mesodermal cells may undergo or chondrogenic differentiation or apoptosis depending whether they are incorporated into the future digital rays or into the interdigital spaces. Both chondrogenesis or apoptosis are induced by local BMPS. However, whereas the chondrogenic-promoting activity of BMPs appears to be regulated through the BMPR 1b receptor, the mechanism by which the BMPs execute the death program remains unknown. The BMP proapoptotic activity requires the expression of members of the msx family of closely related homeobox-containing genes and is finally mediated by caspase activation, but the nature of the caspase(s) directly responsible for the cell death is also unknown. Finally, other growth factors present in the developing autopod at the stages of digit formation such as members of the FGF and TGF beta families modulate the ability of BMPs to induce cell death or chondrogenesis. PMID- 10668470 TI - Oxidative damage, bleomycin, and gamma radiation induce different types of DNA strand breaks in normal lymphocytes and thymocytes. A comet assay study. AB - Most anticancer treatments such as chemo- and radiotherapy induce DNA damage and apoptosis in normal cells. The aim of this study was to assess the induction of single and double DNA strand breaks (ssb and dsb, respectively) and apoptosis in normal human lymphocytes and rat thymocytes subjected to the action of H2O2, bleomycin and ionizing radiation. Normal human peripheral thymocytes and young rat thymocytes were subjected to the following treatments: a) H2O2; b) bleomycin, and c) gamma-radiation, all with various doses. DNA strand breaks were studied with the alkaline and neutral comet assay for detection of ssb and dsb. Apoptosis was quantified morphologically and with DNA agarose gel electrophoresis. After H2O2 treatment, a dose-dependent increase of ssb was observed. Bleomycin treatment produced a moderate increase of ssb at lower concentrations and a striking increase of dsb at higher concentrations that coincided with the presence of apoptosis and DNA ladders. Gamma radiation initially induced the formation of ssb, and after three hours an increase of dsb in a dose-dependent manner. Apoptosis and DNA laddering appeared only 3 hours post-irradiation. The biomonitoring of DNA damage inflicted by antineoplastic agents can be easily performed with the comet assay and could be useful to monitor and modulate chemo- and radiotherapeutic regimes in cancer patients. PMID- 10668471 TI - Tumor cells utilize multiple pathways to down-modulate apoptosis. Lessons from a mouse model of islet cell carcinogenesis. AB - Apoptosis, the process of programmed cell death, plays a critical role in many normal and pathological (disease) processes. In normal tissues, apoptosis functions in the homeostatic maintenance of proper tissue and organ size by eliminating aged cells to offset the birth of new cells that arise by mitosis. In disease, apoptosis can affect the pathological process is two disparate ways. There are diseases that have too much apoptosis such as autoimmune diabetes and Alzheimer's, or those that have too little apoptosis, such as cancer. This review will focus on the latter and, more specifically, detail and summarize some important lessons learned about apoptosis and cancer from studying a transgenic mouse model of islet cell carcinoma, RIP-Tag, as outlined below. PMID- 10668472 TI - Hyperoxia in cell culture. A non-apoptotic programmed cell death. AB - Here we discuss the morphological features and our current understanding of the pathways involved in non-apoptotic cell death from O2 toxicity. Preliminary data on hyperoxic signaling indicate that NF-kappa B translocation (and presumptive activation) is not a result of the p42/p44 MAPK pathway, but a likely downstream consequence of activation of the JNK pathway. Our observations suggest the existence of multiple signal transduction pathways in hyperoxia-induced cell death: one involved in the stress response which appears to be NF-kappa B dependent and another in cell death. PMID- 10668473 TI - Hyperoxia-induced cell death in the lung--the correlation of apoptosis, necrosis, and inflammation. AB - Prolonged exposure to hyperoxia causes tissue damage in many organs and tissues. Since the entire surface area of lung epithelium is directly exposed to O2 and other inhaled agents, hyperoxia leads to the development of both acute and chronic lung injuries. These pathologic changes in the lung can also be seen in acute lung injury (ALI) in response to other agents. Simple strategies to mitigate hyperoxia-induced ALI might not be effective by virtue of merely reducing or augmenting the extent of apoptosis of pulmonary cells. Identification of the specific cell types undergoing apoptosis and further understanding of the precise timing of the onset of apoptosis may be necessary in order to gain a greater understanding of the connection between apoptosis and tolerance to hyperoxia and ALI. Attention should also be focused on other forms of non apoptotic programmed cell death. PMID- 10668474 TI - Apoptosis in coxsackievirus B3-induced myocarditis and dilated cardiomyopathy. AB - Group B coxsackieviruses (CVB), which infect the myocardium, cause myocarditis and dilated cardiomyopathy. However, not all infections of the myocardium result in disease. In the mouse model, CVB infection stimulates autoimmune T cell response to cardiac antigens, and these autoimmune effectors cause myocyte necrosis and cardiomyopathy. Induction of pathogenic autoimmunity depends upon CD4+ Th1 (interferon-gamma positive) cells while Th2 (IL-4 positive) cell responses promote disease resistance. T lymphocytes expressing the gamma-delta T cell receptor (gamma delta +) constitute up to 12% of the inflammatory cells in the heart and are crucial to maintaining a dominant Th1 response phenotype. gamma delta + lymphocytes modulate T cell responses by selectively lysing CD4+ Th2 cells. Th1 cells are not killed by gamma delta + cells. Lysis requires direct cell:cell interaction between the gamma delta + cell and CD4+ Th2 target and is most likely mediated through Fas:FasL interaction. These studies demonstrate a novel mechanism for immune modulation of cytokine responses in vivo. PMID- 10668475 TI - The immune response to apoptotic cells. AB - Programmed cell death (PCD) can be divided into two distinct but linked sequential processes, killing of the cells and removal of the dead cells, which may be a neighboring cell or a professional phagocyte. Following internalization of the apoptotic cell, the phagocyte typically triggers neither the development of a pro-inflammatory response nor the production of autoantibodies directed against apoptotic self antigens. Since apoptotic cells are characterized by translocation of autoantigens such as nucleosomes to the surface of the cell, we tested the hypothesis that excess or abnormally processed apoptotic cells can generate autoantibodies. We have found that syngeneic apoptotic load can induce transient hypergammaglobulinemia, anti-DNA, anticardiolipin, and glomerular depositions in normal mice. Furthermore, we also found that one of the important mechanisms of uptake of apoptotic cells involves opsonization by the complement system, suggesting that deficient states could lead to aberrant handling of apoptotic cells. Therefore, conditions in which apoptotic cells become immunogenic may explain antigen selection in inflammatory and autoimmune conditions, such as in systemic lupus erythematosus (SLE). PMID- 10668476 TI - Programmed cell death as a mechanism of CD4 and CD8 T cell deletion in AIDS. Molecular control and effect of highly active anti-retroviral therapy. AB - Infection with human immunodeficiency virus (HIV) results in the progressive destruction of CD4 T lymphocytes, generally associated with progression of the disease. The progressive disappearance of CD4 T lymphocytes leads to the lack of control of HIV replication and to the development of severe immune deficiency responsible for the occurrence of opportunistic infections associated with AIDS. In this review we discuss premature lymphocyte apoptosis in the context of HIV infection as the consequence of the continuous production of viral proteins, leading to an unbalanced immune activation and to the triggering of apoptotic programs. The chronic immune activation induces the continuous expression of death factors which could turn lymphocytes, including CD4 T cells, CD8 CTL or APC, into effectors of apoptosis, leading to the destruction of healthy activated non-infected cells. Thus, programmed cell death would significantly contribute to peripheral T cell depletion in AIDS, particularly if the Th cell renewal is impaired. Under potent anti-retroviral therapies, a complete normalization of lymphocyte apoptosis is observed, concomitant with a partial restoration of the number and the functions of the immune system. PMID- 10668477 TI - Progress in cancer chemoprevention. AB - More than 40 promising agents and agent combinations are being evaluated clinically as chemopreventive drugs for major cancer targets. A few have been in vanguard, large-scale intervention trials--for example, the studies of tamoxifen and fenretinide in breast, 13-cis-retinoic acid in head and neck, vitamin E and selenium in prostate, and calcium in colon. These and other agents are currently in phase II chemoprevention trials to establish the scope of their chemopreventive efficacy and to develop intermediate biomarkers as surrogate end points for cancer incidence in future studies. In this group are fenretinide, 2 difluoromethylornithine, and oltipraz. Nonsteroidal anti-inflammatories (NSAID) are also in this group because of their colon cancer chemopreventive effects in clinical intervention, epidemiological, and animal studies. New agents are continually considered for development as chemopreventive drugs. Preventive strategies with antiandrogens are evolving for prostate cancer. Anti inflammatories that selectively inhibit inducible cyclooxygenase (COX)-2 are being investigated in colon as alternatives to the NSAID, which inhibit both COX 1 and COX-2 and derive their toxicity from COX-1 inhibition. Newer retinoids with reduced toxicity, increased efficacy, or both (e.g., 9-cis-retinoic acid) are being investigated. Promising chemopreventive drugs are also being developed from dietary substances (e.g., green and black tea polyphenols, soy isoflavones, curcumin, phenethyl isothiocyanate, sulforaphane, lycopene, indole-3-carbinol, perillyl alcohol). Basic and translational research necessary to progress in chemopreventive agent development includes, for example, (1) molecular and genomic biomarkers that can be used for risk assessment and as surrogate end points in clinical studies, (2) animal carcinogenesis models that mimic human disease (including transgenic and gene knockout mice), and (3) novel agent treatment regimens (e.g., local delivery to cancer targets, agent combinations, and pharmacodynamically guided dosing). PMID- 10668478 TI - Preclinical mouse models for cancer chemoprevention studies. AB - To aid in identifying the ability of chemopreventive agents to inhibit tumor development, new preclinical in vivo rodent models have recently been developed. Some of the models contain targeted mutations capable of increasing the incidence and progression of neoplastic lesions, whereas in other models dietary nutrients induce preneoplastic lesions in normal mice. These new preclinical models are assisting the analysis of genetic and environmental factors leading to neoplasia, and clinical studies to evaluate the chemopreventive efficacy of specific nutrients and pharmacological agents. PMID- 10668479 TI - Cellular mechanisms of risk and transformation. AB - Our early work using the first array and imaging methods for the quantitative analysis of the expression of 4000 cDNA sequences suggested that modulation of mitochondrial gene expression was a factor in determining whether colonic epithelial cells displayed a differentiated or transformed phenotype. We have since dissected a pathway in which mitochondrial function is a key element in determining the probability of cells undergoing cell-cycle arrest, lineage specific differentiation, and cell death. Moreover, this pathway is linked to signaling through beta-catenin-Tcf, but in a manner that is independent of effects of the APC gene on beta-catenin-Tcf activity. Utilization of unique mouse genetic models of intestinal tumorigenesis has confirmed that mitochondrial function is an important element in generation of apoptotic cells in the colon in vivo and has demonstrated that modulation of cell death may be involved in intestinal tumor progression rather than initiation. Normal spatial and temporal patterns of cell proliferation, differentiation, and apoptosis in the colonic mucosa are determined by developmentally programmed genetic signals and external signals generated by homo- and heterotypic cell interactions, humoral agents, and lumenal contents. Mitochondrial function may play a pivotal role in integrating these signals and in determining probability of cells entering different maturation pathways. How this is accomplished is under investigation using high density cDNA microarrays. PMID- 10668480 TI - APC and intestinal carcinogenesis. Insights from animal models. AB - The APC protein is a crucial regulator of intestinal cell growth, and mutations in the APC gene are a common initial event in the process of human colorectal carcinogenesis. Animals bearing germline mutations in Apc are therefore important models for human colorectal cancer. These animals have been used both to understand the biology of human colorectal cancer and to screen for agents able to prevent malignant transformation of susceptible intestinal cells. PMID- 10668481 TI - Oncogenic activating mutations in the neu/erbB-2 oncogene are involved in the induction of mammary tumors. AB - Amplification and overexpression of erbB-2/neu is an important determinant in the initiation and progression of human breast cancer. Indeed, transgenic mice that over-express the neu proto-oncogene heritably develop mammary adenocarcinomas. Tumorigenesis in these transgenic strains is associated with activation of the intrinsic catalytic activity of Neu. In many of these tumors, activation of Neu occurs as a result of somatic mutations located within the transgene itself. Examination of the altered neu transcripts revealed the presence of in-frame deletions that encode aberrant Neu receptors lacking 5 to 12 amino acids within the extracellular domain proximal to the transmembrane region of Neu. In addition to these deletion mutants we have also detected single point mutations within this juxta-transmembrane region. The majority of the mutations analyzed affect the one of several conserved cysteine residues present within this region. Introduction of these activating mutations into the wild-type neu cDNA results in its oncogenic conversion. Taken together, these observations suggest that this cysteine-rich region plays an important role in regulating the catalytic activity of Neu. PMID- 10668482 TI - Cyclooxygenase-deficient mice. A summary of their characteristics and susceptibilities to inflammation and carcinogenesis. AB - Cyclooxygenase (COX)-1- and COX-2-deficient mice have unique physiological differences that have allowed investigation into the individual biological roles of the COX isoforms. In the following, the phenotypes of the two COX knockout mice are summarized, and recent studies to investigate the effects of COX deficiency on inflammatory responses and cancer susceptibility are discussed. The data suggest that both isoforms have important roles in the maintenance of physiological homeostasis and that such designations as house-keeping and/or response gene may not be entirely accurate. Furthermore, data from COX-deficient mice indicate that both isoforms can contribute to the inflammatory response and that both isoforms have significant roles in carcinogenesis. PMID- 10668483 TI - Inhibition of cyclooxygenase-2 expression. An approach to preventing head and neck cancer. AB - Cyclooxygenase (COX) catalyzes the formation of prostaglandins (PG) from arachidonic acid. A large body of evidence has accumulated to suggest that COX-2, the inducible form of COX, is important in carcinogenesis. In this study, we determined whether (1) COX-2 was overexpressed in squamous cell carcinoma of the head and neck (HNSCC) and whether (2) retinoids, a class of chemopreventive agents, blocked epidermal growth factor (EGF)-mediated activation of COX-2 expression. Levels of COX-2 mRNA were determined in 15 cases of HNSCC and 10 cases of normal oral mucosa. Nearly a 100-fold increase in amounts of COX-2 mRNA was detected in HNSCC. By immunoblot analysis, COX-2 protein was detected in 6 of 6 cases of HNSCC but was undetectable in normal mucosa. Because retinoids protect against oral cavity cancer, we investigated whether retinoids could suppress EGF mediated induction of COX-2 in cultured oral squamous carcinoma cells. Treatment with EGF led to increased levels of COX-2 mRNA, COX-2 protein, and synthesis of PG. These effects were suppressed by a variety of retinoids. Based on the results of this study, it will be important to establish whether newly developed selective COX-2 inhibitors are useful in preventing or treating HNSCC. Moreover, the anticancer properties of retinoids may be due, in part, to inhibition of COX 2 expression. Combining a retinoid with a selective COX-2 inhibitor may be more effective than either agent alone in preventing cancer of the upper aerodigestive tract. PMID- 10668484 TI - The role of COX-2 in intestinal cancer. AB - Cyclooxygenase (COX), the key regulatory enzyme for prostaglandin synthesis is transcribed from two distinct genes. COX-1 is expressed constitutively in most tissues, and COX-2 is induced by a wide variety of stimuli and was initially identified as an immediate-early growth response gene. In addition, COX-2 expression is markedly increased in 85-90% of human colorectal adenocarcinomas, whereas COX-1 levels remain unchanged. Several epidemiological studies have reported a 40-50% reduction in the risk of developing colorectal cancer in persons who chronically take such nonsteroidal anti-inflammatory drugs (NSAIDs) as aspirin, which are classic inhibitors of cyclooxygenase. Genetic evidence also supports a role for COX-2, since mice null for COX-2 have an 86% reduction in tumor multiplicity in a background containing a mutated APC allele. These results strongly suggest that COX-2 contributes to the development of intestinal tumors and that inhibition of COX is chemo-preventative. PMID- 10668485 TI - COX-2 inhibitors. A new class of antiangiogenic agents. AB - The formation of new blood vessels by angiogenesis to provide adequate blood supply is a key requirement for the growth of many tumors. While normal blood vessels expressed the COX-1 enzyme, new angiogenic endothelial cells expressed the inducible COX-2. We evaluated the role of COX inhibitors in the mouse corneal micropocket assay in which angiogenesis is driven by the addition of a Hydron pellet containing basic fibroblast growth factor (bFGF). Neovascular areas were measured with a slit lamp five days after pellet implantation into the corneal stroma. All animals containing implants with bFGF (90 ng) developed intensive areas of neovascularization, whereas the controls implanted with the Hydron pellet alone did not. Indomethacin (a nonselective COX-1/COX-2 inhibitor) and SC 236 (a COX-2-selective inhibitor) inhibited angiogenesis in a dose-dependent manner. Importantly, the indomethacin-treated mice developed severe gastrointestinal toxicity at the efficacious dose of 3 mg/kg/day. By contrast, gastrointestinal lesions were not observed, and platelet COX-1 activity was unaffected, at anti-angiogenic doses of SC-236 (1-6 mg/kg/day). Furthermore, a COX-1-selective inhibitor, SC-560, was ineffective at doses up to 10 mg/kg, a dose that completely blocked platelet COX-1 activity in these mice. SC-236 was also effective in reducing angiogenesis driven by bFGF, vascular endothelium growth factor (VEGF), or carrageenan in the matrigel rat model. Finally, in several tumor models, SC-236 consistently and effectively inhibited tumor growth and angiogenesis. This novel antiangiogenic activity of COX-2 inhibitors indicates their potential therapeutic utility in several types of cancer. PMID- 10668486 TI - Micronutrient deficiencies. A major cause of DNA damage. AB - Deficiencies of the vitamins B12, B6, C, E, folate, or niacin, or of iron or zinc mimic radiation in damaging DNA by causing single- and double-strand breaks, oxidative lesions, or both. The percentage of the population of the United States that has a low intake (< 50% of the RDA) for each of these eight micronutrients ranges from 2% to 20+ percent. A level of folate deficiency causing chromosome breaks occurred in approximately 10% of the population of the United States, and in a much higher percentage of the poor. Folate deficiency causes extensive incorporation of uracil into human DNA (4 million/cell), leading to chromosomal breaks. This mechanism is the likely cause of the increased colon cancer risk associated with low folate intake. Some evidence, and mechanistic considerations, suggest that vitamin B12 and B6 deficiencies also cause high uracil and chromosome breaks. Micronutrient deficiency may explain, in good part, why the quarter of the population that eats the fewest fruits and vegetables (five portions a day is advised) has about double the cancer rate for most types of cancer when compared to the quarter with the highest intake. Eighty percent of American children and adolescents and 68% of adults do not eat five portions a day. Common micronutrient deficiencies are likely to damage DNA by the same mechanism as radiation and many chemicals, appear to be orders of magnitude more important, and should be compared for perspective. Remedying micronutrient deficiencies is likely to lead to a major improvement in health and an increase in longevity at low cost. PMID- 10668487 TI - Calcium and vitamin D. Their potential roles in colon and breast cancer prevention. AB - The geographic distribution of colon cancer is similar to the historical geographic distribution of rickets. The highest death rates from colon cancer occur in areas that had high prevalence rates of rickets--regions with winter ultraviolet radiation deficiency, generally due to a combination of high or moderately high latitude, high-sulfur content air pollution (acid haze), higher than average stratospheric ozone thickness, and persistently thick winter cloud cover. The geographic distribution of colon cancer mortality rates reveals significantly low death rates at low latitudes in the United States and significantly high rates in the industrialized Northeast. The Northeast has a combination of latitude, climate, and air pollution that prevents any synthesis of vitamin D during a five-month vitamin D winter. Breast cancer death rates in white women also rise with distance from the equator and are highest in areas with long vitamin D winters. Colon cancer incidence rates also have been shown to be inversely proportional to intake of calcium. These findings, which are consistent with laboratory results, indicate that most cases of colon cancer may be prevented with regular intake of calcium in the range of 1,800 mg per day, in a dietary context that includes 800 IU per day (20 micrograms) of vitamin D3. (In women, an intake of approximately 1,000 mg of calcium per 1,000 kcal of energy with 800 IU of vitamin D would be sufficient.) In observational studies, the source of approximately 90% of the calcium intake was vitamin D-fortified milk. Vitamin D may also be obtained from fatty fish. In addition to reduction of incidence and mortality rates from colon cancer, epidemiological data suggest that intake of 800 IU/day of vitamin D may be associated with enhanced survival rates among breast cancer cases. PMID- 10668488 TI - Preclinical and early human studies of calcium and colon cancer prevention. AB - Colorectal cancer continues to be a major cause of tumor mortality in the United States and other countries; despite attempts to improve the screening of high risk populations, the incidence of this disease is still very high. Therefore, chemoprevention continues to be an important goal for the primary prevention of colorectal cancer. Among recent chemopreventive approaches, the administration of calcium and vitamin D continue to be evaluated in both preclinical and clinical studies. Many experimental findings described below have indicated associations between high calcium and vitamin D intake and decreased risk for colorectal cancer. PMID- 10668489 TI - Studies of calcium in food supplements in humans. AB - Colon cancer is one of the commonest cancers in the Western world. Environmental factors appear to predominate as exemplified by a change in incidence in colon cancer within 20 years when people emigrate from a low- to a high-incidence country. It had been suggested that a diet high in energy, fat, and meat content and low in fiber content is most likely responsible. Epidemiologic observations have pointed to a potential effect of calcium or/and vitamin D in reducing the incidence of colon cancer. Other studies have shown a reduction in preneoplastic colon adenomas with increased calcium or/and vitamin D intake. High fat diets were shown to be accompanied by an increase in fecal fatty acids and bile acids or a change in bile acid composition. Soluble fatty acids and bile acids then could interact with the colonic epithelium inducing cell damage and increased proliferation. A hypothesis was developed suggesting that calcium supplementation and increased calcium in the colonic lumen would precipitate these bile acids and fatty acids. Examination of the effect of supplemental calcium or calcium in dairy foods showed a major reduction in fecal bile acids and fatty acids in solution in volunteers and accompanied by a reduction in cytolytic activity. Studies then were performed in patients at risk for colon cancer seeking a change in proliferative biomarkers of risk from a high-risk to a low-risk pattern with supplemental calcium administration. These studies generally have shown a beneficial effect of the addition of calcium at 1.2-2 gm per day in addition to a regular diet for periods of 2 to 6 months. A recently published study also demonstrated that a diet, in which low-fat dairy foods containing an average of about 825 mg of calcium, significantly improved proliferative biomarkers as well as two differentiation bio-markers of risk for colon cancer from a high- to a low risk pattern. These observations, together with recent studies showing reduced adenomatous polyp recurrence when supplemental calcium was provided, demonstrate the potential of calcium and perhaps vitamin D as chemopreventive agents for colorectal neoplasia. PMID- 10668490 TI - Calcium supplements and colorectal adenomas. Polyp Prevention Study Group. AB - Experimental and observational findings suggest that calcium intake may protect against colorectal neoplasia. To investigate this hypothesis, we conducted a randomized, double-blind trial of colorectal adenoma recurrence. Nine hundred thirty patients with a recent history of colorectal adenomas were randomly given calcium carbonate (3 gm daily; 1200 mg elemental calcium) or placebo, with follow up colonoscopies one and four years after the qualifying examination. The main analysis focused on new adenomas found after the first follow-up endoscopy, up to (and including) the second follow-up examination. Risk ratios of at least one recurrent adenoma and ratios of the average numbers of adenomas were calculated as measures of calcium effect. There was a lower risk of recurrent adenomas in subjects assigned calcium. Eight hundred thirty-two patients had two follow-up examinations and were included in the main analysis; the adjusted risk ratio of one or more adenomas was 0.81 (95% CI 0.67 to 0.99); the adjusted ratio of the average numbers of adenomas was 0.76 (95% CI 0.60 to 0.96). Among subjects who had at least one follow-up colonoscopy, the adjusted risk ratio of one or more recurrent adenomas was 0.85 (95% CI 0.74 to 0.98). The effect of calcium seemed independent of initial dietary fat and calcium intake. No toxicity was associated with supplementation. These findings indicate that calcium supplementation has a modest protective effect against colorectal adenomas, precursors of most colorectal cancers. PMID- 10668491 TI - Breast cancer prevention by antiestrogens. AB - A new era has been entered with the first demonstration that an antiestrogen can prevent breast cancer. In a landmark study tamoxifen was shown to reduce the incidence of breast cancer by approximately 50%. The reduction was observed in pre- and postmenopausal women at increased risk of breast cancer. Invasive cancers were reduced, the reduction being in the estrogen receptor-positive cancers. No preventive effect was observed for estrogen receptor-negative tumors. In situ cancers were also significantly reduced. A collateral benefit was a significant reduction in fractures due to osteoporosis. Adverse effects included a very small increase in the incidence of endometrial cancer, cataracts, and stroke. The benefits appear to outweigh the risks for those at high risk. Preliminary studies of a new selective estrogen receptor modulator (SERM 2), raloxifene, developed for the prevention of osteoporosis, have shown that the breast cancer rate was reduced by more than 50% without any concomitant increase in endometrial cancer. The search for a SERM 3, and beyond, may lead to the development of drugs that have the beneficial effects of estrogen while preventing breast cancer and osteoporosis. PMID- 10668492 TI - European trials on dietary supplementation for cancer prevention. AB - European institutions aimed at cancer research and control are spending sizable resources to develop preclinical and clinical chemoprevention trials. Pilot studies showed positive effect on colorectal cell proliferation from supplementation with calcium; vitamins A, C, and E; omega-3 fatty acids; and folic acid. A significant reduction in adenoma recurrence after polypectomy was found in patients randomly assigned to take vitamin A, C, and E supplementation or, to a lesser extent, lactulose. Although first reports showed a disquieting higher incidence of lung cancer in male smokers who took beta-carotene supplementation, the European Organization of Research and Treatment of Cancer (EORTC) planned a chemoprevention study on the prevention of second primary tumors in patients with curatively treated head and neck or lung cancer (EUROSCAN). Retinol palmitate or N-acetylcysteine or both are given for two years. The European Cancer Prevention Organization (ECP) is carrying out a clinical trial in patients with previous adenomas of the large bowel, to test the efficacy of calcium or fiber supplementation on adenoma recurrence. ECP in collaboration with EURONUT has also started a multinational intervention study of the effect of H. pylori eradication and/or dietary supplementation with vitamin C on intestinal metaplasia. PMID- 10668493 TI - Update from Asia. Asian studies on cancer chemoprevention. AB - In Asia, nontoxic dietary products are considered desirable primary prevention vehicles for conquering cancer. As early as 1978, investigators in Korea carried out extensive long-term anticarcinogenicity experiments using the mouse lung tumor model and observed an anticarcinogenic effect of Panax ginseng C.A. Meyer extract in 1980. The results showed that natural products can provide hope for human cancer prevention. A newly established nine-week medium-term model using mouse lung tumors (Yun's model) could confirm the anticarcinogenicity of ginseng that varies according to its type and age. Subsequently, the ginseng was shown by epidemiological studies to be a nonorgan-specific cancer preventive agent associated with a dose-response relationship. The anticarcinogenic effects of vegetarian foods common at every dining table in Korea and some synthetics were also studied using Yun's nine-week model. In brief, ascorbic acid, soybean lecithin, capsaicin, biochanin A, Ganoderma lucidum, caffeine, and a novel synthetic 2-(allylthio)pyrazine decrease the incidence of mouse lung tumors, whereas fresh ginseng (4 years old), carrot, spinach, Sesamum indicum, beta carotene, and 13-cis retinoic acid do not. This result regarding beta-carotene is consistent with the ineffective findings of the ATBC trial, the CARET trial, and the Physicians' Health Study. In 1983, a cancer chemoprevention study group was first established in Japan. Subsequently, (-)-epigallocatechin gallate, cryptoporic acid E, and sarcophytol A from natural products, and synthetic acyclic retinoid and canventol were shown to be anticarcinogenic or chemopreventive in human subjects. Despite the frequent consumption of tea wordwide as a beverage and current experimental evidence of anticarcinogenesis, including controversial results of epidemiological studies, more systematic clinical trials for confirmation of preventive activity of tea against cancer are needed. Placebo-controlled intervention trials of dietary fiber are under study in Japan. In the past decade, new triterpenoids were isolated from various natural sources, and its biological activities were investigated in Asia. In the late 1970s a comprehensive chemoprevention program was established at the Institute of Materia Medica, Chinese Academy of Medical Sciences. Since then, many retinoid compounds have been synthesized and screened in the search for chemopreventive cancer agents. The National Cancer Institute (USA) and China are jointly engaged in the two-nutrition intervention in Linxian, China. The results of joint study of the general population and of dysplasia in China should stimulate further research to clarify the potential benefits of micronutrient supplements. We need to clarify if there is a connection between the lower rates of cancer mortality in Korea and the frequent consumption of anticarcinogenic vegetables or traditional foods, including ginseng and Ganoderma lucidum. The constituents of the nontoxic stable dietary products promise to be the future hope for conquering cancers in the coming years. PMID- 10668494 TI - Squalene, olive oil, and cancer risk. Review and hypothesis. AB - Epidemiologic studies of breast and pancreatic cancer in several Mediterranean populations have demonstrated that increased dietary intake of olive oil is associated with a small decreased risk, or no increased risk, of cancer, despite a high overall lipid intake. Experimental animal models in high dietary fat and cancer also indicate that olive oil either has no effect, or a protective effect, on the prevention of a variety of chemically induced tumors. As a working hypothesis, it is proposed that the high squalene content of olive oil, as compared to other human foods, is a major factor in the cancer-risk reducing effect of olive oil. Experiments in animal models suggest a tumor-inhibiting role for squalene. A mechanism is proposed for the tumor-inhibitory activity of squalene based on its known strong inhibitory activity of HMG-COA reductase catalytic activity in vivo, thus reducing farnesyl pyrophosphate (FPP) availability for "prenylation" of ras oncogene, which relocates this oncogene to cell membranes and is required for the signal-transducing function of ras. Reduction of mutated ras oncogene activation may be useful in breast and colon cancer and may be particularly applicable to pancreatic cancers that are strongly associated with ras oncogenes. PMID- 10668495 TI - Multifunctional aspects of the action of indole-3-carbinol as an antitumor agent. AB - Previous studies from this laboratory have suggested that 2-hydroxyestrone is protective against breast cancer, whereas the other principal metabolite, 16 alpha-hydroxyestrone, and the lesser metabolite quantitatively, 4-hydroxyestrone, are potent carcinogens. Attempts to directly decrease the formation of the 16 hydroxylated metabolite were either unsuccessful or required such high levels of the therapeutic agent as to be impractical. On the other hand the concentration of the protective metabolite, 2-hydroxyestrone, proved to be readily modulated by a variety of agents, both in the direction of increased protection and the opposite direction, increased risk by a variety of agents and activities. We have focussed our attention on indole-3-carbinol, a compound found in cruciferous vegetables, and its further metabolites in the body, diindolylmethane (DIM) and indolylcarbazole (ICZ), because of its relative safety and multifaceted activities. It has been shown that it induces CyP4501A1, increasing 2 hydroxylation of estrogens, leading to the protective 2-OHE1, and also decreases CyP1B1 sharply, inhibiting 4-hydroxylation of estradiol, thereby decreasing the formation of the carcinogenic 4-OHE1. In addition to these indirect effects as a result of altered estrogen metabolism, indole-3-carbinol has been shown to have direct effects on apoptosis and cyclin D, resulting in blockage of the cell cycle. In addition to its antitumor activity in animals, it has also been shown to be effective against HPV-mediated tumors in human patients. All of these responses make the study of its behavior as a therapeutic agent of considerable interest. PMID- 10668496 TI - Resveratrol inhibits cyclooxygenase-2 transcription in human mammary epithelial cells. AB - A large body of evidence suggests that inhibiting cyclooxygenase-2 (COX-2), the inducible form of COX, will be an important strategy for preventing cancer. In this study, we investigated whether resveratrol, a chemopreventive agent found in grapes, could suppress phorbol ester (PMA)-mediated induction of COX-2 in human mammary and oral epithelial cells. Treatment of cells with PMA induced COX-2 mRNA, COX-2 protein, and prostaglandin synthesis. These effects were inhibited by resveratrol. Nuclear runoffs revealed increased rates of COX-2 transcription after treatment with PMA, an effect that was inhibited by resveratrol. Resveratrol inhibited PMA-mediated activation of protein kinase C and the induction of COX-2 promoter activity by c-Jun. Phorbol ester-mediated induction of AP-1 activity was blocked by resveratrol. These data are likely to be important for understanding the anticancer and anti-inflammatory properties of resveratrol. PMID- 10668497 TI - Modification of dietary habits (Mediterranean diet) and cancer mortality in a southern Italian village from 1960 to 1996. PMID- 10668498 TI - Season-specific correlation between dietary intake of fruits and vegetables and levels of serum biomarkers among Chinese tin miners at high risk for lung cancer. PMID- 10668499 TI - BRCA 1/2 gene mutation testing-based cancer prevention and the moral concerns of different types of patients. PMID- 10668500 TI - Cohort analysis of etiological factors for colorectal cancer following endoscopic resection of colorectal tumors. PMID- 10668501 TI - Negative growth regulation of oncogene-transformed human breast epithelial cells by phytochemicals. Role of apoptosis. PMID- 10668502 TI - Fecal pH from patients with colorectal tumors. PMID- 10668503 TI - Oncogenetic information in the hands of physicians and the preventive options of persons who are not their patients. PMID- 10668504 TI - Mammalian Cu-containing amine oxidases (CAOs): new methods of analysis, structural relationships, and possible functions. AB - This thesis describes new and original experimental results on Cu-dependent amine oxidases (CAOs), which show that these enzymes can be conveniently and specifically detected in situ using a peroxidase-coupled activity staining method with 4-Cl-1-naphtole as hydrogen donor substrate. Even more sensitive in situ detection can be achieved using a chemiluminescence-based coupled peroxidase assay which was applied to show that human placenta CAO activity is confined to maternal vessels. A general purification scheme for CAOs is described, and applied to purification of different CAOs. Peptide maps and immunological crossreactivity studies with monoclonal antibodies raised against the purified enzymes showed that they were closely related. Amino acid sequence data for the bovine serum CAO showed that they form a separate group (E.C. 1.4.3.6) with no homology to other enzymes. A cDNA sequence was obtained on the basis of the amino acid sequence data, and this was found to encode a bovine lung CAO, related to bovine serum CAO. The genes for bovine lung and bovine serum CAO are characterized, and Southern blotting analysis of bovine chromosomal DNA shows the existence of a least one more bovine CAO. The purification of human neutrophil CAO is attempted, but it is described how lactoferrin, a protein with many properties in common with CAOs, and with a low degree of sequence identity can account for many observations on human neutrophil CAO. The products of bovine serum CAO oxidation of polyamines are characterised, and 3-aminopropanal is found to be the principal aminoaldehyde produced. Finally, a polyamine-stimulated binding of human placenta CAO to single-stranded DNA is described, and it is reported that the DNA-bound CAO is enzymically active and that the oxidation of DNA-bound polyamines leads to degradation of DNA. In addition to the experimental results, the properties of polyamines and Cu-dependent amine oxidases are reviewed. The polyamines spermidine and spermine interact specifically with nucleic acids and several other molecules. They are synthesised from putrescine, which is a key regulatory molecule formed from ornithine by ornithine decarboxylase, a highly inducible and regulated enzyme. The polyamines can be converted to putrescine by CAOs or spermidine/spermine acetyltransferase and polyamine oxidase. Putrescine is degraded by CAOs, which are also involved in degradation of histamine, a mediator of inflammatory processes. CAOs catalyse the general reaction: R1CH2NHR2 + O2 + H2O-->R1CHO + R2NH2 + H2O2 and in addition to the catabolism of putrescine and histamine CAOs are involved in regulation of growth and apoptosis by to the generation of aminoaldehydes and hydrogen peroxide which have growth inhibitory properties. Several homologous CAOs have been purified and characterized and they form a family with two subgroups. They are homodimers with a relative molecular weight of 180,000 and contain Cu2+ and a modified tyrosine, topaquinone, in the active site. CAOs are present in most tissues with highest amounts in intestine, kidneys, liver and placenta, but the cellular distributions and functions of CAOs are still poorly described, partly due to the use of many different assays and partly due to a broad substrate specificity of the enzymes. However, polyamines and CAOs seem to form a universal system contributing to regulation of growth, differentiation, and apoptosis. PMID- 10668505 TI - [Posttraumatic syringomyelia]. AB - The improvement of preclinical emergency medicine, better surgical and conservative therapies, and the development of intensive care units and specialized centers have improved the survival rate for patients with serious spinal cord injuries. Therefore, more sequelae of chronic spinal cord injuries such as post-traumatic spinal cord cavitations also occur. The first such case was described by Bastian in 1867. Generally, these cavitations were diagnosed from 2 months up to 32 years after the trauma. The overall prevalence of post traumatic syringomyelia (PTS) is not known; however, with the increasing use of magnetic resonance imaging (MRI), its diagnosis has increased, ranging from 2.3% of paraplegic and tetraplegic patients in 1976 and 3.2% in 1985, to nearly 50% in a selected group of patients in 1991 and 1993. In 1995, a 4.45% incidence was reported. In our clinic we are currently treating 440 cases of syringomyelia, 140 of which are PTS. Several observations suggest more than one potential mechanism for the evolution of a post-traumatic cyst or PTS. Various factors, such as hemorrhage or, in particular, ischemia within the spinal cord, blockage of the cerebrospinal fluid (CSF) pathways around the cord or localized meningeal fibrosis either alone or in combination with other factors, may be involved. Clinically, sensory disturbances, loss of motor function, pain, and modification of the deep tendon reflexes are observed in most patients. On MRI, PTS is seen as a longitudinal, cystic cavity within the spinal cord, giving a hypointense signal on T1-weighted images and a hyperintense signal on T2-weighted images. For treatment planning it is mandatory to identify the lower and upper end of the PTS on the MRI. PMID- 10668506 TI - [CINE-MRT for the study of the effects of regional left ventricular wall motion disorders on global heart function after a myocardial infarct and revascularization]. AB - PURPOSE: To determine changes in global cardiac function and mass caused by infarct-associated regional wall motion abnormalities and to compare the changes after revascularization in patients with and without improvement of regional contractility. MATERIALS AND METHODS: 21 patients with regional left ventricular wall motion abnormalities and associated coronary artery stenoses requiring revascularization were examined with a Cine FLASH-2D sequence 26 +/- 12 days after their first myocardial infarction and re-examined three months after revascularization. Regional contractility and volumes and masses of both ventricles were determined. RESULTS: After revascularization, regional wall motion improvement led to decreased left ventricular volumes and improved ejection fractions, whereas patients with persisting wall motion defects showed unchanged left ventricular functional parameters. Comparing both groups of patients, the patients with improvement of regional contractility revealed lower end-systolic volumes and higher ejection fractions at follow-up. Cardiac masses and right ventricular parameters were not different, patients with a depressed right ventricular ejection fraction showed improvement at follow-up. DISCUSSION: After myocardial infarction, revascularization of the infarct-related coronary artery leads to an improvement of left ventricular function only if there is also an improvement of regional contractility. An effect on right ventricular function was not observed three months after the first small- or middle-sized myocardial infarction. PMID- 10668507 TI - [The value of magnetic resonance tomography (MRT) for evaluating ventricular and anastomotic functions in patients with an extra- or intracardiac total cavopulmonary connection (TCPC)-modified Fontan operation]. AB - PURPOSE: To evaluate different MR methods (ventricle and flow measurements) for the postoperative follow-up of hemodynamics in patients with extra- or intracardial TCPC. MATERIALS AND METHODS: Twenty-eight consecutive patients (14 female, 14 male) within the ages of two to thirty-eight years were examined using a 1.5 T Gyroscan ACS-NT scanner (Philips, Best, Netherlands). 7 patients had an extracardial (eTCPC), and 21 an intracardial (iTCPC) tunnel. The calculation of the ventricular function and muscle mass was performed using "multislice multiphase" technique by summing up the end-diastolic and end-systolic areas; the flow measurements were evaluated by phase shift velocity mapping in the superior vena cava (SVC), inferior vena cava (IVC), right (RPA) and left (LPA) pulmonary artery. Besides peak and mean velocity, the mean and maximal flow volumes (ml/min) were calculated. RESULTS: Ejection fraction (EF) of the functionally single ventricle was within the normal range (mean 57%) in 22/28 patients while mean muscle mass was elevated in the group with eTCPC (mean 121 g/m2). The mean flow volumes and the peak velocities in all vessels were higher in the group with iTCPC as compared to the one with eTCPC. Clinically relevant retrograde flows in the IVC were only found in the group with iTCPC (7/21), as well as a significant predominant flow distribution towards the RPA (p < 0.05; Wilcoxon signed-rank test); in the group with eTCPC towards the LPA (n.s.). CONCLUSIONS: MRI is a useful method for the assessment of ventricular function and muscle mass in the follow-up after the modified Fontan operation. MRI flow measurements additionally provided clinically relevant information about the hemodynamics in Fontan patients. PMID- 10668508 TI - [Contrast medium-enhanced MR angiography of the pelvic and leg vessels with an automated table-feed technique]. AB - PURPOSE: To evaluate contrast enhanced magnetic resonance angiography (ceMRA) with an automated table-feed technique in patients with arterio-occlusive disease for imaging of the pelvic and peripheral arteries. METHODS: Twenty-two patients underwent three-dimensional gadolinium-enhanced MR angiography in a three-step automatic table-feed technique on a Magnetom Symphony operating at 1.5 Tesla. Maximum intensity projection images (MIP) were generated from the subtracted and original studies. Image quality and venous contrast were evaluated by two groups of observers. 304 vessels (17 patients) were compared with DSA as the standard of reference. RESULTS: All examinations were performed without any technical problems. Diagnostic quality of the MIP of subtracted data sets was superior to that of the unsubtracted images. Venous overlay was 61% in the lower leg. In a total of 599 observations, a sensitivity of 96% (95%, 82%) and a specificity of 87% (88%, 99%) were high compared to DSA in the detection of significant stenoses > or = 50% (> or = 75%, occlusions). Interobserver correlation was good (linear correlation 0.9). CONCLUSION: Stepping-table digital subtraction contrast enhanced MRA is a promising technique in the diagnosis of peripheral arterio occlusive disease. PMID- 10668509 TI - [The optimization of MR cholangiopancreatography]. AB - PURPOSE: To improve the image quality of magnetic resonance cholangiopancreatography (MRCP) by modification of examination conditions. MATERIALS AND METHODS: MRCP of 72 patients was performed with a 1.5 T system (Magnetom Vision, Siemens, 25 mT/m) using two breath-hold techniques, half fourier acquisition with multislice T2-WI HASTE in MIP technique, and single shot T2-WI turbo-spin-echo (RARE) with different slice thicknesses. The effects of n butylscopolamine were assessed. Furthermore, oral contrast agents [barium sulfate, Fe(II)-gluconate, Fe(II,III)-oxide] in various concentrations were used. The slice thickness was varied for the RARE sequence (3-7 cm). RESULTS: N butylscopolamine had no influence on image quality. Improvements could be attained by variation of the slice thickness. A significant reduction of disturbing background noise was obtained by oral application of iron gluconate, or iron oxide-containing contrast media. Similar improvements were achieved with barium sulfate. CONCLUSIONS: Variation of slice thickness allows an improvement of MRCP quality. Oral contrast media improve the image quality of MRCP. The expense of contrast media may be a determinant of choice. PMID- 10668511 TI - [Fat-saturated, contrast-enhanced spin-echo sequences in the magnetic resonance tomographic diagnosis of peritoneal carcinosis]. AB - PURPOSE: To evaluate contrast-enhanced, fat-saturated spin echo sequences for the detection of peritoneal carcinosis with MRI. MATERIALS AND METHODS: 61 patients, 35 with and 26 without peritoneal carcinosis, were examined with abdominal MRI. Fat-saturated, T1-weighted spin echo sequences were performed before and after administration of Gd-DTPA. In addition, 22 patients with peritoneal carcinosis were examined with contrast-enhanced abdominal CT. RESULTS: 32 of 35 patients with peritoneal carcinosis demonstrated contrast enhancement of the visceral and 30 of 35 enhancement of the parietal peritoneum (91 and 86%, respectively). Wall thickening of the intestine or parietal peritoneum were noted in 21 and 20 of 35 patients (60 and 57%, respectively), ascites in 18 of 35 patients (51%). False positive contrast enhancement of the peritoneum was noted in 4 of 26 patients (15%). In the direct comparison of MRI and CT, 22 of 22 patients versus 7 of 22 patients showed contrast enhancement of the visceral peritoneum (100 and 32%, respectively). For other signs of peritoneal carcinosis (e.g., ascites, peritoneal seedings), no differences in diagnostic reliability were demonstrated. CONCLUSIONS: The use of fat-saturated, spin echo sequences facilitates the diagnosis of peritoneal carcinosis by artifact reduction and improved detection of peritoneal contrast enhancement. MRI with fat-saturated sequences was superior to CT. PMID- 10668510 TI - [Portal contrast medium-enhanced spiral computed tomography of the liver--the correlation of radiological and intraoperative findings and the evaluation of resectability]. AB - PURPOSE: To evaluate the accuracy of spiral computed tomography during arterial portography (SCTAP) in the detection, localization, and resectability of liver tumors in a correlative study between radiology and intraoperative findings. METHOD AND MATERIALS: Retrospectively, SCTAP images of 168 consecutive patients before liver tumor resection were analyzed. The SCTAP studies (100 ml lopromid 300 by automated injector with a flow of 3 ml/s; slice thickness, table feed and reconstruction index 5 mm each; scan-delay 30 s; 120 kV; 250 mAs) were evaluated for the detection, localization, and resectability of focal liver lesions by three experienced radiologists in consensus and were correlated with histopathological and intraoperative findings where available (59/168). RESULTS: The sensitivity of SCTAP for the detection of liver tumors was 91% for all lesions and 84% for lesions < 1 cm. The specificity was only 19% due to a high rate of false-positive lesions (30%) and preselection effects. Typical pitfalls in false positive lesions were inhomogeneous liver perfusion near the portal vein, the falciform ligament, and the gallbladder (19/42). In 30% of the patients SCTAP correctly diagnosed inoperability, in 23% the intraoperative tumor expansion was larger than expected from SCTAP images, which would have changed operability. CONCLUSION: The SCTAP has a high sensitivity in the detection and localization of liver tumors and is a valuable method in the preoperative diagnostic procedure. The method is limited by many false-positive lesions often due to inhomogeneous liver perfusion and the insufficient evaluation of local tumor spread. Therefore, SCTAP should be replaced by MRI in the near future. PMID- 10668512 TI - [The quantification of the magnetization transfer contrast (MTC) effect by calculating MT quotients: does it yield additional information for the differentiation of benign and malignant diseases of the locomotor apparatus?]. AB - PURPOSE: To investigate the potential information of the amount of magnetization transfer effect in musculoskeletal lesions and to compare MT ratios from benign and malignant musculoskeletal lesions. MATERIAL AND METHOD: 49 patients with malignant tumors (3 osteosarcoma, 3 malignant fibrous histiocytoma, 4 chondrosarcoma, 2 Ewing sarcomas) and benign lesions (8 chondroma, 2 fibrous dysplasia, 3 osteoid-osteoma, 6 ganglion cyst, 3 cyst, 3 osteomyelitis, 4 tendinitis, 3 rotator cuff tear, 5 scar tissue) were scanned using routine MRI protocols including T1- and T2-weighted spin echo as well as T2*-weighted gradient echo (FFE) sequences at 1.5 Tesla (ACS II, Philips Medical). Additionally MTC images were generated by combining the FFE sequence and the off resonance MT technique (-1500 Hz off-resonance frequency, 1770 degrees flip angle and 50 ms pulse duration). MT ratios were calculated as Slo-Slm/Slo. RESULTS: The MT ratio of benign lesions was 26 +/- 15%, that of malignant lesions was 22 +/- 6%. The difference was statistically not significant. As expected muscle showed a high MT ratio of 50 +/- 8%. Scar tissue demonstrated an MT ratio of 39 +/- 16% which was significantly higher than the tumor MT ratios. CONCLUSION: MTC (MT ratios) failed to show significant differences between benign and malignant lesions as was expected due to basic differences in cellularity, rate of mitosis and chromatin content. MTC might however gain more importance in separating scar tissue from recurrent tumor in the future. PMID- 10668513 TI - [The evaluation of experimentally induced injuries to the upper cervical spine with a digital x-ray technic, computed tomography and magnetic resonance tomography]. AB - PURPOSE: To compare digital X-ray, CT, and MRI in the evaluation of ligamentous and osseous lesions in upper cervical spine specimens after artificial craniocervical injury with the findings of macroscopic preparation. MATERIALS AND METHODS: A rotation trauma of defined severity was applied to 19 human corpses. After dissection of the neck specimens, digital X-ray (DIMA Soft P41, Feinfocus), conventional and helical CT (CTi, High Speed, GE, collimation 1 mm; pitch 1.0), and MRI were performed from the skull base to C3. The findings were correlated with the macroscopic results of preparation. MR (Magnetom Vision, Siemens) imaging was obtained with a 1.5 T system using 2D- and 3D-sequences. RESULTS: Preparation revealed 6 fractures of the vertebral bodies, 5 fractures of the dens axis, 1 fracture of the arcus anterior of the atlas, 4 osseous flakes at the occipital condylus, and 6 lesions of the alar ligaments. Digital radiography showed all fractures and 4 osseous flakes at the occipital condylus. With conventional and helical CT, all fractures and all ruptured alar ligaments could be detected. 2D MRI depicted 9 of the fractures and 3D MRI showed fractures. With 2D MRI, 2 of the 4 osseous flakes at the condylus could be detected and with 3D MRI one occipital condylus fracture could be depicted. Ligamentous injuries were visualized by 2D MRI in 2 of 6 cases and by 3D MRI in one case. CONCLUSIONS: In post-mortem studies, CT was superior to MRI in the visualization of osseous and ligamentous injuries after trauma of the upper cervical spine. However, these results are not transferable to patients with rotation injury in general. PMID- 10668514 TI - [Direct in-vitro measurement of ultrasound velocity in carcinomas, mastopathic tissue, fatty tissue and fibroadenomas of the female breast]. AB - PURPOSE: The study aimed to investigate ultrasound velocity (SV) in carcinomas, fibrocystic changes, fibroadenomas and fatty tissue of the female breast by means of direct in-vitro measurements. We intended to test whether or not differences in SV exist between the various types of tissue and whether the SV is a useful criterion to differentiate the different tissues. METHOD: SV was measured by comparing transmission time of the ultrasound beam through the specimen and through water. Altogether 40 specimens (12 cancer, 14 fibrocystic changes = FCD, 10 fatty tissues, 3 fibroadenomas, and 1 mixed tissue) were analysed. RESULTS: Velocity differed significantly between fat (1478.5 +/- 6.5 m/s) and tumor (1523.1 +/- 5.9 m/s) (p approximately 10(-11)) and between fat and FCD (1526.0 +/ 9.0 m/s) (p approximately 10(-12)). No significant differences and much overlap were seen between the ultrasound velocities of tumors and FCD. Ultrasound velocity in fibroadenomas (1533.2 +/- 3.8 m/s) was comparable with that in carcinomas and FCD. CONCLUSIONS: We conclude that ultrasound velocity may add complementary information to echogenicity (B-scan). Thus, a locally exact correlation of echogenicity and sound velocity might allow for an improved tissue characterization. PMID- 10668515 TI - [The in-vitro evaluation of different embolectomy catheters for the treatment of acute pulmonary embolism]. AB - INTRODUCTION: Pulmonary embolism (PE) is one of the most common cardiovascular diseases and frequently causes death. As a rule, PE is treated with thrombolytic therapy or surgical thrombectomy. MATERIALS AND METHODS: In an in vitro model of the right lung, we tested four different percutaneous transluminal thrombectomy devices: a pigtail-catheter with an angled 3-cm (40 degrees) distal tip, the clot buster, the hydrolyser catheter, and a modified hydrolyser. In 16 consecutive and repetitive experiments fresh thrombi were inserted and we evaluated the effectiveness of the system with respect to time, fragment size, reduction of the Miller score, and handling. RESULTS: Mean intervention times of the catheter systems were 23 min (pigtail), 14.4 min (modified hydrolyser), 13.8 min (clot buster), and 10.8 min (hydrolyser). The maximum size of the produced fragments range from 0.5 to 3.5 mm by the pigtail and from 0.5 to 1 mm by the other systems. The Miller score reduction was from 14.4 to 2.8 (pigtail), 13.8 to 1.8 (clot-busters), 14.6 to 1.2 (hydrolyser), and 16.4 to 1 (modified hydrolyser). DISCUSSION: All four catheter systems were effective in the treatment of pulmonary embolism. The pigtail catheter is the most simple system but more time consuming and less effective in the fragmentation of emboli and reduction of the Miller score compared to the other three catheter systems. These systems were comparable in our model but especially the handling of the hydrolyser was encouraging. PMID- 10668516 TI - [A venous aneurysm in the mediastinum as a diagnostic error possibility in computed tomography]. PMID- 10668517 TI - [Spontaneous thoracic soft-tissue emphysema in a plain-film image. A radiological approach in a more than unusual etiology]. PMID- 10668518 TI - [Pseudoaneurysm of the A. epigastrica inferior after laparoscopic cholecystectomy]. PMID- 10668519 TI - [Nonparasitically induced renal lymph reflux with chyluriae]. PMID- 10668520 TI - [A triple ureter with an ectopic opening into the vagina]. PMID- 10668521 TI - Early aggressive DMARD therapy: the key to slowing disease progression in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an autoimmune disease characterized by joint inflammation, joint destruction, progressive disability, and premature death. Patients at risk for poor prognoses can be identified by a variety of prognostic indicators. These include sociodemographic factors (e.g., older age, female sex), clinical indicators (e.g., higher joint counts), laboratory variables (e.g., higher erythrocyte sedimentation rate, high rheumatoid factor titer), and radiographic indicators (e.g., the presence of bone erosions). Patients with a poor prognosis, as evidenced by the presence of one or more indicators of poor outcome, should be treated promptly and aggressively with disease-modifying antirheumatic drugs (DMARDs) or combination DMARD therapy to limit or prevent further disease progression. Limiting the severity of RA with early and aggressive treatment is the best way to minimize the dire consequences of untreated or inadequately treated disease. PMID- 10668522 TI - Disease modification in rheumatoid arthritis with leflunomide. AB - Rheumatoid arthritis (RA) is characterized by chronic inflammation and irreversible destruction of articular cartilage and bone. Disease progression as assessed by radiographic imaging of structural joint damage is a key outcome measure in RA. Joint damage is especially rapid during early phases of RA, thus the current trend of early aggressive therapy with disease-modifying antirheumatic drugs (DMARDs). Radiographic analysis of disease progression with the novel DMARD leflunomide was compared to methotrexate and sulfasalazine in two large, placebo-controlled, randomized Phase III studies (N = 580). The results as indicated by changes in x-ray scores indicate that leflunomide and both active comparators slow disease progression significantly better than placebo (P < or = 0.01). Slowing of disease progression with leflunomide was similar to sulfasalazine at 6 months but better than methotrexate (P < or = 0.049) at 12 months. These data verify the ability of leflunomide to slow disease progression and confirm its disease-modifying potential. PMID- 10668523 TI - Efficacy and safety of the new DMARD leflunomide: comparison to placebo and sulfasalazine in active rheumatoid arthritis. AB - The efficacy and safety of the novel DMARD leflunomide was compared to placebo and sulfasalazine in a randomized, double-blind study. At Week 24, leflunomide significantly reduced tender and swollen joint counts and physician and patient assessment scores compared to placebo (P < 0.001). Response rates with leflunomide were significantly greater than placebo: ACR 20% (55% vs 29%, P = 0.0001). Comparable response rates were observed with sulfasalazine (ACR 20%: 56%). Leflunomide significantly improved HAQ scores compared to placebo or sulfasalazine (P < 0.009). The onset of action with leflunomide was rapid and was seen as early as Week 2. Radiographic disease progression was significantly slower with leflunomide than placebo (P < 0.01). Leflunomide was well tolerated. No long-term safety issues were reported with leflunomide in patients who opted to continue treatment for up to 2 years. Efficacy of leflunomide in the treatment of RA was maintained at 2 years. PMID- 10668524 TI - Leflunomide improves quality of life in rheumatoid arthritis. AB - Functional disability in rheumatoid arthritis (RA) interferes with activities of daily living and severely affects patient quality of life. It results in increased levels of work disability and high medical costs. A new goal of RA therapy is to reduce or prevent functional disability. Patients' perception of overall health status in RA was assessed using several instruments (HAQ, MHAQ, SF 36, and PET) in Phase III double-blind, placebo-controlled, randomized trials comparing the new DMARD, leflunomide to sulfasalazine and methotrexate. Leflunomide significantly improved patient quality of life compared to placebo in both the European (P = 0.0001) and North American (P = 0.0001) studies. Reduction in HAQ scores with leflunomide (-0.50 vs -0.29; P = 0.0086) was significantly greater than sulfasalazine. Leflunomide also significantly reduced MHAQ scores versus methotrexate (-0.29 vs -0.15; P < or = 0.05). These changes were seen as early as Week 4. These results highlight the efficacy of leflunomide in RA therapy. PMID- 10668525 TI - Leflunomide versus methotrexate: a comparison of the European and American experience. AB - This paper compares and contrasts the results of two major Phase III clinical trials that compared the efficacy and safety of leflunomide, a new disease modifying antirheumatic drug (DMARD), and methotrexate. In both the American trial (US301) and the multinational trial (MN302), patients with active rheumatoid arthritis (RA) were given either leflunomide (20 mg/day after a loading dose of 100 mg/day for 3 days) or methotrexate (7.5-15 mg/week) for 52 weeks. US301 was also placebo-controlled. Folate supplementation was mandatory in US301 but was given to < 10% of the patients in MN302. In US301, American College of Rheumatology (ACR) 20% response rates and improvement in tender and swollen joints were significantly better than placebo in both treatment groups, but were not significantly different from each other. Both treatments significantly retarded radiographically assessed progression of RA compared to placebo, but the degree of retardation was significantly greater with leflunomide. In MN302, the ACR response rate and improvement in tender and swollen joints with leflunomide were similar to those seen in US301. The ACR response rate and improvements in all efficacy variables with methotrexate were significantly greater than with leflunomide, however. Radiographically assessed disease progression was not statistically different with the two treatments. Use of methotrexate without folate in MN302 was associated with a higher incidence of clinically significant elevations of liver enzyme levels. These results indicate that both leflunomide and methotrexate are effective DMARDs. The symptomatic relief provided by both drugs is similar when they are given with folate supplementation. PMID- 10668527 TI - What is your diagnosis? Oblique fracture of the odontoid process from the axial body. PMID- 10668526 TI - Thoughts on the transmission of Sarcocystis neurona. PMID- 10668528 TI - A veterinarian's recourse for libelous language. PMID- 10668529 TI - Compendium of animal rabies prevention and control, 2000. The National Association of State Public Health Veterinarians. PMID- 10668530 TI - Mortality rates and causes of death among emaciated cats. AB - OBJECTIVE: To determine mortality rates and causes of death for thin (i.e., lean or emaciated) cats and, if mortality rates were high, to determine factors associated with risk that cats would be thin. DESIGN: Cohort study. ANIMALS: 1,138 cats examined at 27 private veterinary practices in the northeastern United States. PROCEDURE: Body condition of the cats was scored (emaciated, lean, optimally lean, optimal, heavy, obese) between 1991 and 1992. Follow-up information on whether cats had developed any illnesses, whether cats had died, and, if cats had died, cause of death was obtained between 1994 and 1996. Mortality risk for emaciated cats was estimated, using cats in optimal condition as the reference group. RESULTS: Survival curves for emaciated cats were significantly lower than those for cats of other body conditions. Compared with cats in optimal condition, emaciated cats were 4.4 times as likely to die during the follow-up period. However, after adjusting for age and excluding cats that died within 1 year after body condition was scored, emaciated cats were no longer significantly more likely to die. Emaciated cats were more likely to die of an unknown cause than were cats of optimal condition. Risk factors for emaciated body condition included preexisting illness, age, and Siamese breed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that emaciated cats had a significantly higher risk of death, compared with cats in optimal body condition. Serious illness and advancing age accounted for much, and perhaps all, of this increased risk of death. PMID- 10668531 TI - Ophthalmomyiasis interna anterior associated with Cuterebra spp in a cat. AB - An 8-year-old domestic shorthair cat was examined for severe anterior uveitis of the right eye that was unresponsive to aggressive treatment with anti inflammatory drugs and for a possible intraocular parasite or foreign body trapped within a large fibrin clot in the anterior chamber. Surgical exploration of a presumed entry site on the caudal aspect of the third eyelid led to keratotomy and removal of a larval parasite later identified as a first instar Cuterebra spp. Aggressive treatment with anti-inflammatory drugs was continued after surgery, and intraocular pressure was monitored closely to ensure that the cat did not develop glaucoma. Two weeks after surgery, the cat had vision in the affected eye, with resolving uveitis and a normal fundus. Six weeks after surgery, the uveitis continued to resolve; however, the cat did not have vision in the affected eye, and examination of the fundus revealed retinal atrophy. In contrast to the condition in humans, a Cuterebra spp larval infection within the eye of cats may cause not only an intense, acute inflammatory reaction, but also retinal degeneration and blindness despite prompt surgical removal. PMID- 10668532 TI - Retrobulbar pseudotumor of the orbit in a cat. AB - Idiopathic nonspecific inflammatory disease of the orbit (orbital pseudotumor) was diagnosed detected in a cat. The cat had progressive lagophthalmia, keratitis, and decreased motion of the right eye. Four months later, the left eye was affected in a similar manner. Response to antibiotics and immunosuppressive agents was not detected. Computed tomography of the brain and orbits revealed bilateral thickening of the sclera and episcleral tissues. Bilateral exenteration of the eyes was required because of worsening clinical signs or corneal perforation. Histologic examination revealed proliferation of spindle cells and fibrovascular tissue within and adjacent to the sclera. PMID- 10668533 TI - Bacteria associated with pyothorax of dogs and cats: 98 cases (1989-1998). AB - OBJECTIVE: To determine the organisms most commonly isolated from pleural fluid from dogs and cats with pyothorax. DESIGN: Retrospective study. ANIMALS: 51 dogs and 47 cats. PROCEDURE: Results of bacteriologic culture of pleural fluid samples obtained by means of thoracentesis were obtained from medical records. To obtain information on in vitro antimicrobial susceptibility of organisms commonly isolated from dogs and cats, records of all dogs and cats examined during 1998 were reviewed, and information was obtained on identity and in vitro antimicrobial susceptibility of aerobic organisms isolated from samples other than urine or urinary tract samples. RESULTS: Median ages of dogs and cats were 4 years. Bacteria were isolated from pleural fluid samples from 47 of 51 (92%) dogs and 45 of 47 (96%) cats. Obligate anaerobic bacteria were isolated from 28 dogs and 40 cats. A mixture of obligate anaerobic and facultative bacteria was isolated from 17 dogs and 20 cats. Samples from cats most often yielded a member of the nonenteric group (most commonly members of the genus Pasteurella), whereas those from dogs more often yielded a member of the family Enterobacteriaceae (most commonly E coli). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that antimicrobial agents chosen for the initial treatment of dogs and cats with pyothorax should be active against a mixture of obligate anaerobic and facultative bacteria. PMID- 10668534 TI - Influence of open surgical correction on intermediate-term outcome in dogs with subvalvular aortic stenosis: 44 cases (1991-1998). AB - OBJECTIVE: To compare outcome and intermediate-term survival for dogs undergoing open surgical correction of subvalvular aortic stenosis (SAS) with those for dogs with SAS that did not undergo surgery. DESIGN: Retrospective study. ANIMALS: 44 dogs with congenital SAS. PROCEDURE: Maximum instantaneous systolic pressure gradients were determined by use of Doppler echocardiography. Cardiopulmonary bypass and open surgical correction of SAS (membranectomy with or without septal myectomy) was performed in 22 dogs, whereas 22 dogs did not undergo surgical correction. Cumulative survival was compared between surgical and nonsurgical groups, using Kaplan-Meier nonparametric analysis and a Mantel-Cox log-rank test. RESULTS: Initial systolic pressure gradients were not significantly different for dogs undergoing surgery (128 +/- 55 mm Hg), compared with those that did not undergo surgery (117 +/- 57 mm Hg). Systolic pressure gradients were significantly decreased after surgery in dogs that underwent surgery (54 +/- 27 mm Hg). Cumulative survival was not significantly different between dogs in the surgical and nonsurgical groups. Censoring surgery-related mortality in the analysis still did not reveal a significant difference in cumulative survival between the surgical and nonsurgical groups. CONCLUSIONS AND CLINICAL RELEVANCE: Despite reductions in the systolic pressure gradient and possible associated improvement in exercise tolerance, a palliative benefit on survival was not documented in dogs undergoing surgery for SAS. PMID- 10668535 TI - Complications associated with the use of indwelling epidural catheters in dogs: 81 cases (1996-1999). AB - OBJECTIVE: To evaluate complications associated with use of indwelling epidural catheters in dogs in a clinical setting. DESIGN: Retrospective clinical study. ANIMALS: 81 client-owned dogs. PROCEDURE: Medical records were reviewed for dogs in which a 19-gauge epidural catheter was placed percutaneously at L7-S1 and advanced to the point of maximum efficacy for pain control (between L7 and T4, depending on the procedure). Catheters were used to provide perioperative epidural analgesia during surgeries that included perineal (n = 6), hind limb (33), abdominal (43), thoracic (5), forelimb (2), and cervical (1) procedures. RESULTS: Catheters were maintained in situ from 1 to 7 days (mean, 2.3 days; median, 2.0 days). Sixty-four dogs did not have complications; 17 dogs had minor complications. Catheter dislodgement was the most common complication (13/80 [16%] dogs). Catheter site contamination without inflammation developed in 2 (2.4%) dogs; inflammation at the catheter site developed in 2 (2.4%) dogs but was not related to duration of time the catheter was in place. Complications were not serious and did not require treatment other than catheter removal. Dogs that dislodged their catheters were significantly younger (mean, 2.9 years; median, 2.0 years) than other dogs (mean, 6.2 years; median, 6.0 years). Dogs that received femoral fracture repair dislodged their catheters more often (62.5%) than dogs undergoing other procedures (10.9%). CONCLUSIONS AND CLINICAL RELEVANCE: The complication rate associated with temporary epidural catheterization of dogs appears to be low, and complications generally are not serious. PMID- 10668537 TI - Productivity characteristics of high-performing commercial swine breeding farms. AB - OBJECTIVE: To determine productivity characteristics of high-performing swine breeding herds in the United States and to determine associations among number of litters per mated female per year (LMFY), number of pigs weaned per sow (PWS), and lactation duration. DESIGN: Cohort study. SAMPLE POPULATION: 1997 productivity records for 685 herds. PROCEDURE: Herds were ranked on the basis of number of pigs weaned per mated female per year, and herds in the upper 10th percentile of this ranking were designated as high-performing herds. Productivity measurements for these herds were compared with values for the remaining herds. RESULTS: High-performing herds had shorter lactation durations and higher mean breeding female inventories than did the remaining herds. High-performing herds also had better reproductive efficiency and used farrowing facilities more efficiently than did the remaining herds. For the high-performing herds, lactation duration was significantly associated with PWS but was not significantly associated with LMFY. In contrast, for the remaining herds, lactation duration was not significantly associated with PWS but was significantly associated with LMFY. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that high-performing commercial swine farms could increase PWS by improving preweaning mortality rate and number of pigs born alive, but that LMFY was already maximal. For other herds, however, shortening lactation duration would likely decrease farrowing interval and improve efficiency of the reproductive cycle without reducing litter size. PMID- 10668536 TI - Treatment and outcome of dogs with leptospirosis: 36 cases (1990-1998). AB - OBJECTIVE: To characterize serologic and clinical features and outcome of dogs with leptospirosis that were treated conservatively (i.e., medical management alone) or with hemodialysis. DESIGN: Retrospective study. ANIMALS: 36 dogs with leptospirosis. PROCEDURE: History; results of physical examinations, ultrasonography, and serologic, hematologic, and serum biochemical analyses; time to resolution of azotemia; and outcome were obtained from medical records. Dogs were treated conservatively (n = 22) or with hemodialysis (14). RESULTS: Between 1990 and 1998, amount of rainfall was positively correlated with number of cases of leptospirosis identified per year. Serum antibodies against 6 Leptospira serovars were measured, and titers were highest to Leptospira pomona in 16 (44%) dogs, L bratislava in 9 (25%) dogs, and L hardjo in 1 (3%) dog. Eight (22%) dogs had equally high titers to L pomona and L bratislava, 1 (3%) had equally high titers to L grippotyphosa and L canicola, and 1 (3%) had high titers to L grippotyphosa, L pomona, L canicola, and L bratislava. During initial evaluation, all dogs were azotemic. Thirty (83%) dogs survived, including 12 of 14 (86%) dogs treated with hemodialysis and 18 of 22 (82%) treated conservatively. Serum creatinine concentration was similar in both groups after resolution of clinical signs. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with L pomona and L bratislava was recognized as a cause of leptospirosis in dogs, and resulted in development of acute renal failure with various degrees of azotemia. Prognosis for dogs with mild to moderate azotemia was good with conservative treatment, whereas treatment with hemodialysis appeared to improve prognosis for dogs with severe azotemia. PMID- 10668538 TI - Diagnosis and treatment of torsion of the spiral colon in an alpaca. AB - A 14-year-old 61.7-kg (136-lb) alpaca was examined for colic of 24 hours' duration. An exploratory celiotomy was performed because of lack of response to medical treatment and ultrasonography revealed an abnormally large amount of free fluid in the peritoneal cavity. Exploration of the abdomen revealed a 20-cm diameter mass, consisting of most of the ascending colon. The spiral colon was thick and edemetous, and it was decided to resect the spiral colon. Following a few complications, the alpaca was discharged 17 days after surgery. Colic in camelids is considered a severe problem because clinical signs are subtle and often not recognized until the condition is untreatable. Camelids are reported to be stoic animals, and may have few signs of pain despite severe abdominal disease. Alpacas with signs of abdominal pain should undergo early and complete physical, laboratory, and diagnostic imaging evaluations. Rapid identification of the need for surgery is vital for a successful outcome. PMID- 10668539 TI - [Diabetic ketoacidosis presenting as acute abdomen]. AB - Three patients, two women aged 21 and 67 and a man aged 43 years, presented at the emergency department with diabetic ketoacidosis and abdominal symptoms mimicking an acute abdominal condition. In two of them laparotomy was performed which proved to be negative. Abdominal symptoms resolved after correction of metabolic, fluid and electrolyte disturbances. Symptoms indicating a possible diagnosis of acute abdomen have to be regarded as being compatible with diabetic ketoacidosis per se. However, a potential acute abdominal problem prompting surgical intervention should not be overlooked; it may have been the precipitating factor for diabetic ketoacidosis. PMID- 10668540 TI - [Cavernous sinus syndrome]. AB - A clear and concise description and clinical interpretation of the cavernous sinus syndrome are lacking. Pathological changes in or around the cavernous sinus may lead to failure of eye muscle nerves and of one or more branches of the trigeminal nerve. The clinical signs of the cavernous sinus syndrome are combinations of failure of these cranial nerves sometimes with exophthalmus. Because many nerves can be wholly or partially involved in the syndrome, there is no clinical uniformity and the cavernous sinus syndrome has never been well defined. A neurotopographical classification is proposed in order to simplify the multiple interpretations of the cavernous sinus syndrome. The classical cavernous sinus syndrome is divided into three syndromes: the syndrome of the superior orbital fissure, the syndrome of the lateral wall of the cavernous sinus and the central cavernous sinus syndrome. PMID- 10668541 TI - [Endovascular stent implantation as a treatment for iliac artery disease]. AB - A stent is an endovascular prosthesis that may be used in the treatment of intermittent claudication caused by lesions of the A. iliaca communis and the A. iliaca externa in which earlier balloon dilatation has proved insufficiently effective. The expansion is caused by inflation of an angioplasty balloon (plastic remodelling: Palmaz stent) or by self-expansion due to elastic transformation as in the Wail stent or to thermic memory metal, as in the Memotherm stent. Evaluation of the literature shows that stent placement is a safe method of treatment. The proportion of initial technical success appears to be higher than that of balloon angioplasty, especially in the treatment of total occlusions. The haemodynamic situation immediately after treatment also appears to be better in case of stent placement. Long-term comparison of the clinical efficacy is not well possible because the published studies differ with regard to patient population, definition of indication and criteria of success. PMID- 10668542 TI - [Critical review of anti-influenza drugs]. AB - Neuraminidase inhibitors such as zanamivir and oseltamivir belong to a new class of antiviral drugs for the treatment and prevention of influenza. As yet however, the therapeutic efficacy of these drugs (shortening of recovery time by approximately one day) has only been demonstrated in healthy adults affected by influenza A, but not in risk groups and in influenza B disease, whereas studies of prophylactic efficacy are still going on. Neither do these drugs impact on viral spread, a public health risk against which the economic advantages of early work resumption have to be weighed. Since flu symptoms can be caused by other germs than the influenza A or B virus, caution in prescribing these drugs seems warranted, also to prevent the development of drug resistance. In addition, when designing therapeutic efficacy trials in risk groups, selecting the rate of secondary complications and death may be more adequate as clinical endpoint than (economically important) duration of illness. PMID- 10668543 TI - [Measles epidemiology in the Netherlands: a exploratory analysis of notification]. AB - OBJECTIVE: Explorative analysis of the effects of vaccination policy on measles incidence. DESIGN: Retrospective study and mathematical modelling. METHOD: Analysis of national and regional case notifications of measles provided by the Inspectorate of Health in the Netherlands over the period from January 1976 (when vaccination was started) through September 1999. Also computer simulations with a mathematical epidemic model of measles were used to calculate the incidence of measles from 1976 onwards. RESULTS: According to the model results, measles should not persist with the current vaccination programme. However, the case notification data showed that measles appeared to persist at a nation-wide level. At a regional level, measles did not persist, not even in regions with low vaccine coverage. A possible cause of the unexpected persistence at the national level is the asynchronous regional course of the 6-year epidemic cycle of measles, where measles infection 'jumps' from one region to the other. PMID- 10668544 TI - [Children with stumbling gait due to acute spinal cord compression]. AB - Three previously healthy children, two girls aged 2 and almost 5 years and a boy aged 20 months, developed a progressively stumbling gait within days. In two this occurred after a period of weeks during which they complained of, or seemed to have back pain. In all three cases acute spinal cord compression by a malignant tumour was diagnosed. Histological examination revealed Ewing sarcoma, granulocytic sarcoma and T-cell lymphoma. Surgical decompression led to complete neurological recovery. Although rare, acute spinal cord compression during childhood is a medical emergency because of the risk of neurological morbidity. Back pain, weakness and a stumbling gait usually are the first symptoms. Sensory symptoms and sphincter dysfunction may develop later. Early recognition is essential, as prognosis depends on neurological findings and duration of symptoms when treatment is started. PMID- 10668545 TI - [Transmission of hepatitis B virus by a surgeon]. PMID- 10668546 TI - [Transmission of hepatitis B virus by a surgeon]. PMID- 10668547 TI - [Transmission of hepatitis B virus by a surgeon]. PMID- 10668548 TI - [Clinical thinking and decision making in practice. An elderly patient with vertigo and high sedimentation rate]. PMID- 10668549 TI - [In-vivo analysis of nitric oxide]. PMID- 10668550 TI - [Analysis of extracellular substances with microdialysis]. PMID- 10668551 TI - [Change in expression pattern of myosin heavy chain isoforms in soleus muscle of the rats after hindlimb suspension]. PMID- 10668552 TI - [Effect of 2,5-hexanedione on myosin heavy chain isoform content in perineal muscles of the rats]. PMID- 10668553 TI - The painted Amsterdam anatomy lessons: anatomy performances in dissecting rooms? AB - The Anatomy Lesson of Dr. Nicolaes Tulp, painted by Rembrandt in 1632, has recently been fully restored. From 02-10-98 to 10-01-99 this painting and some other Amsterdam painted anatomy lessons were exhibited in the Mauritshuis in The Hague, with the title "Rembrandt under the scalpel". The unique Tulp painting is one of those portraits painted in the tradition of the famous group portraits which flourished in 17th-century Holland, a predominantly urban, middle-class society where the main patrons of the arts were the leading citizens of the various towns. Moreover, it is a portrait in the tradition of the anatomy lessons especially painted for the Guild of Surgeons for their Guild Room. Nine such lessons have been painted for the guild and are still to be found in Dutch museums (Mauritshuis and Amsterdam Historical Museum). The anatomy lesson of Prof. Andreas Bonn, dated 1792, as well as some group portraits of the leading persons of the guild also play an important role in the Amsterdam group portraits. In 1925 the Amsterdam anatomist Louis Bolk commissioned Martin Monnickendam to paint another anatomy lesson. The restoration of the painting of Dr. Tulp has provided new information concerning the original composition of Rembrandt and the later additions. However, from an anatomical point of view, it is doubtful whether the Amsterdam anatomy lessons depict a real contemporary anatomical demonstration. They provide, together with archival sources, reliable information about the praelectores anatomiae and the leading persons of the guild, but fail to give much information about the dissecting room, the anatomy theatre or the procedure. The anatomical demonstration procedures of the guild are discussed in relation to the painted anatomy lessons. PMID- 10668554 TI - On the innervation of the donkey testis. AB - The innervation pattern of the adult donkey testis was investigated by immunohistochemistry and acetylcholinesterase histochemistry. Autonomous nerves reach the testis by three access-routes as funicular, mesorchial and caudal contributions. From these, the funicular contribution accompanying the testicular artery and pampiniform plexus is the strongest and most important one. Testicular innervation in the donkey is not uniform. The spermatic cord as well as the epididymal region, cranial and caudal poles (tunica albuginea and adjacent parenchyma and stroma) are well innervated, mostly by vascular nerves. Towards the free border of the testis, the nerve density in the tunica albuginea decreases continuously. In the interior of the gonad, approximately one third of the testis, situated between the free border and the central mediastinum, is practically devoid of any innervation. The great majority of the testicular nerves demonstrated by the present techniques are non-myelinated vascular nerves which react positive for dopamine-beta-hydroxylase and tyrosine hydroxylase, thus representing postjunctional sympathetic fibers. Many of these also contain neuropeptide Y. The testicular innervation of the donkey testis is free of cholinergic fibers. Calcitonin gene-related peptide-containing nerves are found as solitary varicose axons in the wall of blood vessels, but also in stromal connective tissue of the spermatic cord, tunica albuginea and septula testis. PMID- 10668555 TI - Verification of the two-dimensional disector, a method for the unbiased estimation of density and number of myelinated nerve fibers in peripheral nerves. AB - Quantification of the number of myelinated fibers in peripheral nerves is a common requirement in quantitative morphology. This parameter provides important information on the consequences of various physiological, pathological and experimental conditions on the nerve structure and is one of the main indicators of success of peripheral nerve repair. In this paper, the theoretical rationale for the application of stereological principles to obtain unbiased estimates of the density and total number of myelinated fibers in peripheral nerves is discussed and a simple stereological method is described. The method is applied together with a systematic random sampling scheme, that was optimized for the purposes of the present study, and with sampling scheme analysis by calculating the coefficient of error (CE). The stereological method, which consists of a two dimensional variation of the classical disector procedure (two-dimensional disector), and the sampling scheme are verified by comparing estimates with the true density and total number of myelinated fibers in peripheral nerve trunks where true values have been accurately determined by extensive counting. The verification of the 2-D disector method, both of normal and regenerated nerves, showed that estimates of density and total number of myelinated nerve fibers are unbiased. The method also proved to be efficient (time-saving): Estimation of density and total number of myelinated fibers in a single nerve takes about 2-3 hours. PMID- 10668556 TI - Afferent connections of the caudal raphe pallidus nucleus in rats: a study using the fluorescent retrograde tracers fluorogold and true-blue. AB - Afferent connections to the caudal region of the nucleus raphe pallidus (RPa) in rats were studied using fluorogold and true-blue as tracers. Due to its ability to produce limited injection sites, true-blue proved to be more appropriate than fluorogold for studying long distance connections in a narrow structure such as the RPa. Fluorescent, retrogradely-labeled perikarya were found in the preoptic area (median, medial and lateral nuclei), hypothalamus (anterior, dorsal, lateral and posterior areas, and the peri- and paraventricular nuclei), zona incerta, central gray (dorsal, ventral and ventro-lateral), reticular formation of the brainstem, trigeminal spinal nuclei and in the spinal cord (laminae V-X at thoracic, lumbar and sacral levels). This connection pattern suggests the involvement of the RPa in autonomic, somatic and endocrine functions. PMID- 10668557 TI - Angioarchitectural form, functional distributive pattern and classification of the filiform papillae on the crossbred Japanese cat tongue anterodorsal surface in scanning electron microscopic specimens. AB - The purpose of this study was to undertake a three-dimensional comparative observation of the angioarchitectural form, functional distributive pattern and classification of the filiform papillae (FiP) as they appear on the entire dorsal surface of the front of the Japanese cat tongue using microvascular cast specimens (MVCS). By means of the corrosive resin casting technique, the MVSC of the FiP of the cat tongue were prepared and examined in detail under the scanning electron microscope (SEM). On the frontal half of the anterodorsal surface of the anterior tongue, types I-V of the FiP are arranged in the form of a A with the point in the direction of the apex and in an oblique line running from the antero central to both postero-peripheral regions. In the rear half of the anterodorsal surface of the anterior part of the tongue, types I-V of the FiP are arranged in the form of a V with the point in the direction of the root and in an oblique line running from both antero-peripheral regions to the postero-median region or towards the pharynx on the anterior centro-dorsal surface of the tongue. The FiP, arranged in an oblique line running fron the central to the apical part of both the periphery and the pharyngeal region of the cat tongue, can be classified into five types (Types I-V) according to the shape and size of the main process (MP), numbers of the accessory processes (AP) and regional position of the lozenge arrangement. FiP Types I-III consisted of an MP which contained a large spoon shaped and concave network process, and the AP contained a bundle of spin-like processes arranged radially at the anterior basal margin of the MP. FiP Types IV and V consisted only of MP. It was conjectured that the lozenge arrangement of the A and V form FiPs, classified into five types (Types I-V) from the frontal portion of the anterodorsal surface toward the pharynx on the front of the tongue, play a functionally assistant role in the mastication of food and sucking of liquid, including milk. PMID- 10668558 TI - Angioarchitectural form, functional distributive pattern and classification of the fungiform papillae on the crossbred Japanese cat tongue anterodorsal surface in scanning electron microscopic specimens. AB - The purpose of this study was to undertake a three-dimensional comparative observation of the angioarchitectural form, functional distributive pattern, and to classify the fungiform papillae (FuP) found sporadically and geometrically on the oblique lines of the filiform papillae (FiP) on the entire dorsal surface of the crossbred Japanese cat foretongue in microvascular cast specimens (MVCS). By means of the corrosive resin casting technique, the MVCS of the FuP of the cat tongue were prepared and examined in detail under the scanning electron microscope (SEM). With regard to the arrangement of FiPs and FuPs on the frontal half of the antero-dorsal surface of the cat foretongue, FiP types I-V and FuP types I-IV are arranged a A form with the point in the direction of the apex and in an oblique line running from the antero-central portion to both postero peripheral portions. In the rear half of the antero-dorsal surface of the foretongue, FiP types I-V and types FuP I-IV are arranged in a V form with the point in the direction of the radix and in an oblique line running from both antero-peripheral portions to the postero-median portion or towards the pharynx on the anterior centro-dorsal surface of the cat foretongue. With the FuP arranged in the medial zone (M) and peripheral zone (P) running from the central to the apical parts, both the peripheral parts and the pharyngeal can be classified into four types (FuP types I-IV) according to the shape and size of the main process (MP), the number of the accessory processes (AP) and the regional position of the lozenge arrangement. FuP Types I-III consisted of a MP containing a large half oval, network-like process, and the AP contained a bundle of spine-like processes arranged radially in a circle at the anterior basal margin of the MP. The FuP Type IV was made up of only one MP. In the central zone (C) of a lozenge part arranged in the A and V forms, there was no FuP as a taste organ. It was assumed that in the lozenge arrangement of the A and V forms, except for the C zone, FuPs could be classified into four types (FuP types I-IV) from the frontal portion of the antero-dorsal surface toward the pharynx on the foretongue, and would play an assistant functional role in receiving the sense of taste of foods and liquids. PMID- 10668559 TI - Anomalous artery directly connecting the external and internal carotid arteries. AB - An anomalous artery directly connecting the external with the internal carotid artery was encountered on the right side of a 68-year-old Japanese female cadaver. This anomalous artery (5 mm in diameter, 12 mm in length) branched out from the posterior aspect of the external carotid at the level of the origin of the lingual artery, ran obliquely upward posteriorly along the course of the hypoglossal nerve, and was confluent with the anterior aspect of the internal carotid artery. No other variations were found in the morphological aspects of, or in the anatomical relationships between, the carotid arteries and their surrounding structures on either side. The carotid body-like structure was observed at the carotid bifurcation and was innervated by small branches of the glossopharyngeal, the vagus and the sympathetic trunk. Embryologically, it is conceivable that this anomalous artery may have derived from the right second branchial arch artery, although there is no abnormality in other derivative structures of the second pharyngeal arch. There may have been no effect from this anomaly on the functions of the arterial blood flow and blood supply under normal circumstances in the present case, but this report may be of embryological significance and contribute some insight into the mechanisms of the formation of the carotid circulation systems. PMID- 10668560 TI - The relationship between the tibialis posterior tendon and the accessory navicular. AB - Out of a total of 116 cadaver feet, 29 specimens were selected by means of palpation of the tuberosity of navicular for a possible presence of the accessory navicular. They were then radiographed and the accessory navicular was detected in ten. Also three fresh amputation specimens with an accessory navicular were added to the study. A total of 13 legs was dissected and in nine of them, the tibialis posterior tendon inserted directly into the accessory navicular without extending to the sole of the foot. In these feet, the second part of the tibialis posterior tendon originated from the accessory navicular, extending to the normal insertions. There was no connection between these two parts and when traction was applied to either one, no movement was observed in the other. Also a fibrocartilaginous mass was detected in four specimens, probably formed to resist the friction between the tendon and the bone. These results may explain the pronated foot in the presence of the navicular, due to the loss of the function of the tibialis posterior tendon. PMID- 10668561 TI - [Plastination histologic investigations on the inserting pars terminalis aponeurosis dorsalis of three-sectioned fingers]. AB - With the help of thick transparent cross-sections of fingers, manufactured by plastination histology, it has been shown that fibres of the extensor aponeurosis insert not only the way described in current textbook on the basis phalangis distalis and on the capsule of the distal interphalangeal joint. Our studies prove that parts of the dorsal extension plate insert dorsally to the nail matrix and others run over the edge of the basis phalangis to the proximal part of the diaphysis where they intermingle with the periost of the phalanx. PMID- 10668562 TI - Expression of inositol trisphosphate receptors. PMID- 10668563 TI - Basolateral store-operated Ca(2+)-entry in polarized human bronchial and colonic epithelial cells. AB - Bronchial epithelial cells respond to extracellular nucleotides from the luminal and basolateral side activating Cl- secretion via [Ca2+]i increase. In this study we investigated the differences of apically (ap) and basolaterally (bl) stimulated [Ca2+]i signals in polarized human bronchial epithelial cells (16HBE14o-). Specifically we investigated the localization of 'capacitative Ca2+ entry' (CCE). 16HBE14o- cells grown on permeable filters were mounted into an Ussing chamber built for the simultaneous measurement of Fura-2 fluorescence and electrical properties. Application of ATP from both sides induced a rapid [Ca2+]i increase and subsequent sustained [Ca2+]i plateau due to transmembraneous Ca(2+) influx. The use of different nucleotides revealed the following rank order or potency which was very similar for addition from the apical or basolateral side: UTP (EC50 ap: 4 microM, bl: 5 microM) > ATP (EC50 ap: 4 microM, bl: 10 microM) > ADP (n = 4-7 from both sides). 2-MeS-ATP, AMP, adenosine and beta gamma-methylene ATP were ineffective (n = 3 from both sides). The ATP- (ap and bl) induced Ca2+ influx was only abolished by removal of basolateral Ca2+. This was also true for receptor-independent activation of Ca(2+)-influx by intracellular Ca(2+)-store depletion with 2,5 Di-(tert-butyl)-1,4-benzohydroquinone (BHQ) (10 microM). Also in polarized T84 cells the basolateral carbachol and BHQ activated Ca2+ plateau was exclusively sensitive to removal of basolateral Ca2+. We propose that in all polarized epithelial cells the CCE entry pathway is located in the basolateral membrane. We furthermore suggest that Ca2+[i elevating agonists acting from the apical side of the epithelium lead to the opening of a basolateral CCE pathway. PMID- 10668564 TI - The characterization and quantification of antigen-induced Ca2+ oscillations in a rat basophilic leukaemia cell line (RBL-2H3). AB - Using the ratiometric Ca2+ indicator, indo-1, the antigen-induced increase in intracellular Ca2+ concentration ([Ca2+]i) was measured in individual RBL-2H3 cells which had been passively sensitized with monoclonal antibody to the dintrophenyl (DNP) haptenic group. Antigenic stimulation using DNP-human serum albumin conjugate (DNP-HSA) induced concentration-dependent asynchronous Ca2+ oscillations, or irregular spikes. To achieve a quantitative comparison of the effects of different concentrations of antigen on changes in Ca2+[i, the area under the curve (AUC) of Ca2+ oscillations in each cell was calculated. The dose response curve of the calculated AUC is consistent with the bell-shaped dose response curve for antigen-induced mediator release, depolarization and 86Rb(+) efflux. Ca2+ oscillations induced by antigenic stimulation were abolished by removal of external Ca2+ and the subsequent reintroduction of external Ca2+ caused their resumption. To investigate the role of Ca2+ oscillations in the secretory response, changes in [Ca2+]i induced by concanavalin A (Con-A), A23187, thapsigargin and NECA were also monitored. Con-A mimicked the response induced by antigen, whilst A23187 and thapsigargin induced a large transient non-oscillatory response. NECA, an adenosine receptor agonist, induced only a small transient rise in Ca2+[i without oscillatory behaviour. Since all these stimuli accept NECA induced degranulation in these cells, it is suggested that, although Ca2+ oscillations are not essential for the initiation of secretion, they probably underlie the in-vivo physiological response of mast cells and basophils to an antigenic challenge. They also seem to enhance the efficacy of the Ca2+ signal. PMID- 10668565 TI - Construction of a confocal microscope for real-time x-y and x-z imaging. AB - We describe the construction of a simple 'real-time' laser-scanning confocal microscope, and illustrate its use for rapid imaging of elementary intracellular calcium signaling events. A resonant scanning galvanometer (8 kHz) allows x-y frame acquisition rates of 15 or 30 Hz, and the use of mirrors to scan the laser beam permits use of true, pin-hole confocal detection to provide diffraction limited spatial resolution. Furthermore, use of a piezoelectric device to rapidly focus the objective lens allows axial (x-z) images to be obtained from thick specimens at similar frame rates. A computer with image acquisition and graphics cards converts the output from the microscope to a standard video signal, which can then be recorded on videotape and analyzed by regular image processing systems. The system is largely made from commercially available components and requires little custom construction of mechanical parts or electronic circuitry. It costs only a small fraction of that of comparable commercial instruments, yet offers greater versatility and similar or better performance. PMID- 10668566 TI - Effects of hypochlorite-modified low-density and high-density lipoproteins on intracellular Ca2+ and plasma membrane Ca(2+)-ATPase activity of human platelets. AB - The presence of hypochlorite-modified lipoproteins in atherosclerotic lesions suggests that HOCl, a naturally occurring oxidant formed by the myeloperoxidase catalyzed reaction of H2O2 and Cl-, is a candidate for generation of modified lipoproteins in vivo. We have previously demonstrated that Cu(2+)-oxidized LDL inhibits platelet plasma membrane Ca(2+)-ATPase (PMCA) in isolated membranes and causes an increase in cytosolic Ca2+ in resting whole platelets. However, Cu(2+) oxidized LDL may not be identical in structure and function to the physiologically modified lipoprotein. Since platelet function may be affected by native and modified lipoproteins, the effect of HOCl-modified LDL and HDL3 on platelet PMCA and on the free intracellular Ca2+ concentration ([Ca2+]i) of whole platelets has been investigated. We demonstrate that in contrast to Cu(2+) oxidized LDL, HOCl-modified LDL and HDL3 stimulate platelet PMCA activity in isolated membranes and that this effect results in a decrease of [Ca2+]i in vivo. Thus, HOCl-oxidation produces modified lipoproteins with the potential for altering platelet function and with properties different from those of the Cu(2+) oxidized counterparts. PMID- 10668567 TI - Alterations in calcium-mediated signal transduction after traumatic injury of cortical neurons. AB - Calcium influx and elevation of intracellular free calcium ([Ca2+]i), with subsequent activation of degradative enzymes, is hypothesized to cause cell injury and death after traumatic brain injury. We examined the effects of mild-to severe stretch-induced traumatic injury on [Ca2+]i dynamics in cortical neurons cultured on silastic membranes. [Ca2+]i was rapidly elevated after injury, however, the increase was transient with neuronal [Ca2+]i returning to basal levels by 3 h after injury, except in the most severely injured cells. Despite a return of [Ca2+]i to basal levels, there were persistent alterations in calcium mediated signal transduction through 24 h after injury. [Ca2+]i elevation in response to glutamate or NMDA was enhanced after injury. We also found novel alterations in intracellular calcium store-mediated signaling. Neuronal calcium stores failed to respond to a stimulus 15 min after injury and exhibited potentiated responses to stimuli at 3 and 24 h post-injury. Thus, changes in calcium-mediated cellular signaling may contribute to the pathology that is observed after traumatic brain injury. PMID- 10668568 TI - Bladder exstrophy: staged reconstruction. AB - The modern staged approach to bladder exstrophy reconstruction has undergone significant changes since it was first advocated by Jeffs and Cendron in the 1970s. Although varied surgical approaches have been tried in the bladder exstrophy condition, the staged approach to bladder exstrophy repair has withstood the test of time. Progress continues to be made in evaluating the outcome of older types of staged reconstruction, with continuing modification and improvements in the modern approach to staged reconstruction. This treatise will update the reader on recent advances in the treatment of bladder and cloacal exstrophy. PMID- 10668569 TI - Insights into causes of sexual ambiguity. AB - Our understanding of the causes of sexual ambiguity has progressed from the determination of the hormonal etiologies to defining the genetic basis of intersex disorders. The localization of specific genes involved in the process of sexual differentiation has made it possible to determine the mutations and other molecular events that result in sexual ambiguity. With this information, some disorders can now be diagnosed before birth and possibly even treated in utero. PMID- 10668570 TI - Changing concepts in hypospadias repair. AB - After more than 100 years of innovation in hypospadias repair, new concepts regarding penile curvature, the urethral plate, and the means for urethroplasty are changing surgical practices. With the introduction of the tubularized incised plate technique it is possible to simplify decision-making algorithms in hypospadias surgery, while achieving good functional and cosmetic results. PMID- 10668571 TI - Engineering tissues and organs. AB - Genitourinary tissues can be engineered in-vitro and in-vivo for reconstruction using selective cell transplantation in combination with acellular matrices. This technology involves an interdisciplinary approach combining techniques of cell biology and materials sciences towards the development of functional tissues or organs. Tissues and organs in urology, such as the bladder, clitoris, corpus cavermosum, kidney, testis, ureter and urethra have been created in the laboratory, with varying degrees of functionality. Cells have also been recently used in patients as bulking agents for the treatment of vesicoureteral reflux and urinary incontinence. As the science of tissue engineering evolves, one can expect a wider application of this technology to the armamentarium of urologic surgery. PMID- 10668572 TI - Current management of the infant with myelomeningocele. AB - Infants with myelomeningocele continue to be a management dilemma for urologists. This article discusses many of the current questions that surround the newborn with myelomeningocele. What radiologic studies should be performed and when? How does the clinician determine if bladder drainage is adequate or requires altering? If a problem is identified what are the surgical options? When should urodynamics be performed? Ultimately how one manages, follows and initiates treatments in the newborn will have a significant effect on the long-term morbidity seen in this population of children. PMID- 10668573 TI - Andrology, sexual dysfunction, infertility. PMID- 10668574 TI - Role of the urological surgeon in the age of intracytoplasmic sperm injection. AB - In the last few years, it has been demonstrated that intracytoplasmic sperm injection is an effective procedure for treating patients with severe male infertility. Before this technique was available, the urological surgeon involved in the reproduction field dealt mainly with reconstructive (vasoepididimostomy, vasovasostomy) or endoscopic surgery of the seminal duct as well as varicocelectomy. Nowadays, the urological surgeon continues to participate in the field of reproductive surgery by applying methodologies which enable natural conception, and has to be involved in all aspects of sperm retrieval techniques. PMID- 10668575 TI - Oocyte insemination with spermatozoa precursors. AB - Since the use of testicular spermatozoa in programs of assisted fertilization proved very successful, attention was focussed on the use of spermatids also carrying 23 chromosomes. Several difficulties became obvious; the first one concerned the recognition of round spermatids. This is a problem which does not concern elongating and elongated cells. The intra-cytoplasmic injection of elongated spermatids resulted in several pregnancies but this is not so for the round ones. Although, in the group of patients in whom only round spermatids are found at the time of the attempt, is to be divided into two categories; patients in whom previous research allowed to find spermatozoa, however few, and patients who never produced spermatozoa at all. This last group is no longer an indication for intracytoplasmic sperm injection procedure unless in the future new culture media allow a maturation into elongated forms. PMID- 10668576 TI - Pharmacological treatment of erectile dysfunction. AB - There is growing evidence that the field of pharmacotherapy, particularly oral drugs, will be dominant in the future management of sexual dysfunction. Basic research has led to the understanding of the intracellular mechanisms that control penile smooth muscle contractility and therefore erection, opening a vast area for pharmacological intervention. Moreover, the importance of central neurohormonal mechanisms has made these pathways the target for new centrally acting drugs. Given these trends most patients suffering from erectile dysfunction will respond to pharmacological agents in the not so distant future. PMID- 10668577 TI - Pharmacological treatment of premature ejaculation. AB - Ejaculatory dysfunction is the most common male sexual disorder and premature ejaculation the most common presentation of ejaculatory dysfunction. Convincing data are lacking from controlled clinical studies to support sustainable long term efficacy of psychosexual counselling in the management of premature ejaculation. The pharmacological treatment of premature ejaculation is now receiving increased attention from both physicians with an interest in sexual medicine and from the pharmaceutical industry. PMID- 10668578 TI - Female sexual dysfunction: anatomy, physiology, evaluation and treatment options. AB - It has been estimated that up to 76% of women, depending upon their age, have complaints of sexual dysfunction, including decreased libido, vaginal dryness, pain with intercourse, decreased genital sensation and difficulty or inability to achieve orgasm. Female sexual dysfunction is a significant problem that affects the quality of life of many women. This review addresses the etiologies and incidence of female sexual complaints, as well as new findings in the evaluation and treatment of female sexual dysfunction. PMID- 10668579 TI - What's new in Peyronie's disease. AB - Within the past year further scientific studies have been performed to find the aetiology of Peyronie's disease. Advances in the conservative management of the disease by extracorporeal shockwave therapy have been reported in many centres. The operation of plaque excision and grafting, which has been performed for over 25 years, has now been replaced by the superior operation of plaque incision with venous patch. PMID- 10668580 TI - Bibliography. Current world literature. Pediatric urology. PMID- 10668581 TI - Bibliography. Current world literature. Andrology, sexual dysfunction, infertility. PMID- 10668582 TI - Evaluation of six serological tests in diagnosis and postoperative control of pulmonary hydatid disease patients. AB - Latex agglutination (LA), passive hemagglutination (PHA), immunoelectrophoresis (IEP) and specific IgE, IgM, IgG enzyme-linked immunosorbent assay (ELISA) tests for diagnosis and postoperative follow-up of 79 patients with surgically confirmed pulmonary hydatidosis were evaluated. Specific IgG ELISA was the most sensitive test (83.5%) and the least sensitive tests were specific IgE ELISA (44.3%) and IEP (50.6%). The specificity obtained for all the serologic test was above 97% in all cases. The greatest number of false positives in all tests (except IEP) occurred in patients with Taenia saginata and Taenia solium cysticerci infestations and in patients with lymphoma and leukemia. Specific IgG ELISA demonstrated the highest negative predictive value (93.8%). No statistically significant differences (p > 0.050) were found in the sensitivity of the tests when patients with only one cyst and patients with various cysts were compared. Considering only the patients without relapse, the percentage of seropositive patients increased in all tests at 1 and 3 months after surgery. After that time the percentage of seropositive patients decreased. At 48 months after surgery all patients without relapse became negative in IEP, specific IgE ELISA, and specific IgM ELISA. The antibody titers in all seropositive patients increased during the 3 months after surgery. From these 3 months onward, antibody levels decreased in all serologic tests studied in the group of patients without relapse. The patients who had relapses during the first year after surgery presented persistently elevated antibody titers in all postoperative sera. The antibody titers of the patients who relapsed between the third and fourth years after surgery decreased progressively the third month after surgery, and increased in the serum obtained at the moment of relapse diagnosis. Our results show that persistence of elevated antibody titers in patients with pulmonary hydatidosis in the year after surgery or titer increase after a progressive decrease are indicative of relapse or reinfection. PMID- 10668583 TI - Cefepime versus ceftazidime for the treatment of serious bacterial infections. AB - The objective of this open, comparative and randomized clinical trial was to compare the safety and efficacy of cefepime and ceftazidime in the treatment of adults with severe infections. Fifty patients were included; 25 received cefepime (1 g b.i.d.) and 25 ceftazidime (1 g t.i.d.). Demographic characteristics of the groups were similar. The drugs were well tolerated and adverse reactions were minor and comparable in both groups. Forty-four patients were evaluated (22 from each group). There was a satisfactory clinical response in 86% and 77% in the cefepime and ceftazidime groups, respectively. In patients with microbiologically documented infection 83% (15/18) treated with cefepime and 64% (9/14) treated with ceftazidime responded satisfactorily (no statistical significance). In conclusion, the data confirms that cefepime is as safe and effective as ceftazidime for the treatment of serious bacterial infections, with the advantage of being administrated only twice daily. PMID- 10668584 TI - Long-term non-progression of HIV-1 in a patient coinfected with HTLV-II. AB - A 37-year-old man coinfected with HIV-1 and human T-lymphotropic virus type II presumably through injection drug use had a high CD4+ count and low HIV viral load without anti-retroviral therapy for over six years. As an HIV long-term non progressor, his case supports the hypothesis that coinfection with HTLV-II does not adversely affect the course of HIV disease. PMID- 10668585 TI - Evaluation of the RapID Yeast Plus System for the identification of yeast. AB - We evaluated the RapID Yeast Plus System using 117 fresh and frozen clinical yeast isolates. The Uni-Yeast-Tek System was used to establish the correct identification. The Vitek System was used as the arbiter for any discrepant results, along with morphology. Of 117 isolates tested, the RapID Yeast Plus System identified 96.6% correctly. The RapID Yeast Plus System is an accurate and reliable alternative to other commonly used yeast identification systems. PMID- 10668586 TI - In vitro evaluation of cefepime and other broad-spectrum beta-lactams for isolates in Malaysia and Singapore medical centers. The Malaysia/Singapore Antimicrobial Resistance Study Group. AB - The degree of activity of several beta-lactam antimicrobial agents was assessed in Malaysia (four medical centers) and Singapore (two medical centers) tested against 570 clinical isolates. The organisms were tested locally by the Etest (AB BIODISK, Solna, Sweden) method, validated by concurrent use of quality assurance strains (94.1% accurate performance overall). Ten groups of bacteria were tested against cefepime, cefpirome, ceftazidime, ceftriaxone, piperacillin/tazobactam, oxacillin, and imipenem. Among the tested Escherichia coli and Klebsiella spp., the occurrence of extended spectrum beta-lactamase-producing phenotypes was 5.6 7.0% and 36.7-38.0%, respectively. These strains remained most susceptible (97.5 100.0%) to cefepime and imipenem. Ceftazidime-resistant Enterobacter spp. (21.4% resistant), Citrobacter spp. (15.0%), indole-positive Proteus spp. (6.0%), and Serratia spp. (9.7%) were not resistant to cefepime, and only one strain was resistant to imipenem. Imipenem was generally most potent against non fermentative Gram-negative bacilli such as Acinetobacter spp. and Pseudomonas aeruginosa. All tested beta-lactams were active against the oxacillin-susceptible staphylococci, except ceftazidime (MIC90, 12 micrograms/mL; 63.2-84.8% susceptibility rates). Overall spectrums of activity (rank by % resistance) favored imipenem (3.5%) > cefepime (7.7%) > cefpirome (8.9%) > piperacillin/tazobactam (13.2%) > ceftriaxone (14.7%) > ceftazidime (16.9%). No significant differences in resistance patterns were noted between monitored nations, and these results indicate emerging, elevated rates of resistance versus the studied broad-spectrum beta-lactams in Malaysia and Singapore. Results provide benchmark data for future studies using quantitative methods to determine antimicrobial resistance in these geographic areas. PMID- 10668587 TI - In vitro evaluation of cefepime and other broad-spectrum beta-lactams against bacteria from Indonesian medical centers. The Indonesia Antimicrobial Resistance Study Group. AB - The in vitro activity of cefepime and six other broad-spectrum beta-lactams (cefpirome, ceftazidime, ceftriaxone, imipenem, piperacillin/tazobactam (4 micrograms/mL fixed concentration), and oxacillin was evaluated against 191 isolates of clinical bacteria from Indonesia. Susceptibility testing was performed using Etest (AB BIODISK, Solna, Sweden) methodology. Isolates from 10 species groups were selected for analysis: Escherichia coli, Klebsiella spp., Enterobacter spp., indole-positive Proteae, Serratia spp., Acinetobacter spp., Pseudomonas aeruginosa, and oxacillin-susceptible staphylococci. The overall rank order of spectrum of activity was (% resistant): imipenem (2.2%) > cefepime (7.3%) > piperacillin/tazobactam > cefpirome > ceftazidime > ceftriaxone (16.2%). The "fourth-generation" cephalosporins, cefepime and cefpirome, displayed greater activity compared with the "third-generation" cephalosporins, ceftazidime, and ceftriaxone, against the 60 E. coli and Klebsiella spp. (30 each) isolates. Phenotypic extended spectrum beta-lactamase occurrence rates among the E. coli and Klebsiella spp. were 23.3 and 33.3%, respectively. Imipenem, cefepime, and cefpirome inhibited 95.7% of the 46 isolates of inducible Amp C cephalosporinase producing Enterobacteriaceae. The majority of the resistance observed to imipenem and cefepime among tested Indoneisian strains was attributable to the nonfermentative Gram-negative bacilli, P. aeruginosa and Acinetobacter spp. These results indicate the presence of beta-lactam resistance in Indonesia and the need for continued antimicrobial surveillance in this nation and region of the world, preferably using accurate quantitative methods. PMID- 10668588 TI - In vitro evaluation of broad-spectrum beta-lactams in the philippines medical centers: role of fourth-generation cephalosporins. The Philippines Antimicrobial Resistance Study Group. AB - Cefepime is a potent broad-spectrum "fourth-generation" cephalosporin. The in vitro activity of cefepime was compared to that of cefpirome, ceftazidime, ceftriaxone, imipenem, and piperacillin/tazobactam in a multilaboratory (nine medical centers) Philippine surveillance project from March through October 1998. A total of 626 Gram-positive and Gram-negative organisms (10 species groups) were tested by the Etest method (AB BIODISK, Solna, Sweden) with results validated by current quality control strain analysis. The overall rank order of usable spectrum of activity was imipenem (4.2% resistance), cefepime (4.5%), cefpirome (5.0%), piperacillin/tazobactam (5.8%) > ceftriaxone (11.2%) > ceftazidime (15.3%), and results did not differ significantly between medical centers. Ceftazidime-resistant Escherichia coli and Klebsiella spp. occurred at rates of 13.3% and 31.1%, respectively, indicating extended-spectrum beta-lactamase (ESBL) activity. Imipenem (100% susceptible), cefepime, and cefpirome (both > or = 97.8% susceptible) were active in vitro against these ESBL phenotypes. Organisms with ceftazidime and/or ceftriaxone-resistant profiles consistent for hyper-production of Amp C cephalosporinases were detected at high rates among the Citrobacter spp. (29.2%) and Enterobacter spp. (45.8%); however, imipenem (100.0% susceptible) and cefepime (98.9%) remained active. Cefepime and imipenem (both 87.5% susceptible) were the most active agents tested against Acinetobacter spp. whereas piperacillin/tazobactam was most effective against P. aeruginosa (80.0% susceptible). Most tested beta-lactams (except ceftazidime) were active versus oxacillin-susceptible staphylococci. These data should be used as a guide for treatment selection with beta-lactam compounds in the Philippines and to serve as a resistance benchmark in comparisons with future studies in this nation. PMID- 10668589 TI - In vitro evaluation of cefepime and other broad-spectrum beta-lactams in Taiwan medical centers. The Taiwan Antimicrobial Resistance Study Group. AB - The rates of resistance to commonly used antimicrobial agents have been documented to be at alarmingly high levels in Taiwan for both Gram-positive and Gram-negative species. This study was conducted to assess the current resistance patterns in six medical centers strictly controlled using a common MIC methodology and quality assurance measures. Cefepime, a new clinically introduced broad-spectrum "fourth-generation" cephalosporin, was compared to other members in this class including ceftazidime, cefpirome, ceftriaxone, piperacillin/tazobactam, and imipenem. These antimicrobials were tested against ten species groups of common clinical isolates of Enterobacteriaceae, non-enteric Gram-negative bacilli, and oxacillin-susceptible Staphylococcus spp. The results confirmed that extended spectrum beta-lactamase (ESBL) production in Klebsiella spp. (21.7%) and Escherichia coli (16.7%) was common in all medical centers surveyed. Cefepime was more active against these two species as well as against Amp C producing species, indole-positive Proteae, and Acinetobacter species. The activity of cefepime was comparable although slightly less than that of ceftazidime against Serratia spp. and Pseudomonas aeruginosa strains. All or nearly all staphylococci isolates were susceptible to the beta-lactam antimicrobial agents, except for ceftazidime. Overall, these antimicrobial agents had descending spectrums of activity as follows: imipenem > cefepime > cefpirome > piperacillin/tazobactam > ceftazidime > ceftriaxone for the 550 isolates tested. Cefepime seems to be an important broad-spectrum beta-lactam that can be used with confidence against many important pathogens in Taiwan, including those harboring resistance mechanisms. A continued surveillance program seems prudent for this geographic area. PMID- 10668590 TI - In vitro evaluation of cefepime and other broad-spectrum beta-lactams in 22 medical centers in Japan: a phase II trial comparing two annual organism samples. The Japan Antimicrobial Resistance Study Group. AB - An antimicrobial resistance surveillance study in Japan is presented representing the second year (Phase II) results from 22 medical centers. Each participant laboratory tested (Etest, AB BIODISK, Solna, Sweden) 100 organisms, 10 strains each from 10 species groups including Escherichia coli, Klebsiella spp., Enterobacter spp., Citrobacter spp., indole-positive Proteae, Serratia spp., Acinetobacter spp., Pseudomonas aeruginosa, and oxacillin-susceptible Staphylococcus aureus and coagulase-negative staphylococci. Generally only modest variations in the activity of the studied broad-spectrum beta-lactams was observed compared to the study a year before. Specifically, extended spectrum beta-lactamase (ESBL) rates in E. coli increased (2.9 to 8.1%), but the ESBL rate in Klebsiella spp. fell (8.6 to 5.0%). Overall the resistance to the beta-lactams varied from a 4.7% decrease (ceftazidime as a consequence of a modified staphylococcal breakpoint criteria) to a 1.0% increase (cefepime, not significant). The rank order of spectrums in 1998 only changed for cefoperazone sulbactam (6.1% resistance) that was active against more strains than cefpirome (6.8% resistance). The overall spectrum rank order for the 1998 Japan sample (% resistance) was: cefepime (3.2%) > imipenem (4.1%) > cefoperazone-sulbactam (6.1%) > cefpirome (6.8%) > ceftazidime (8.4%) > piperacillin (19.9%). As with a similar study in 1997, imipenem-resistant isolates of P. aeruginosa and Serratia spp. were discovered with metalloenzymes, usually found in the same medical centers. These results demonstrate the continued in vitro activity and potential sustained clinical efficacy of several broad-spectrum beta-lactams in Japan. Rapid emergence of new or novel resistance were not wide spread using a precise quantitative MIC system. Continued surveillance in this nation would be prudent to document the activity of this clinically valuable class of safe, antimicrobial agents. PMID- 10668591 TI - In vitro evaluation of broad-spectrum beta-lactams tested in medical centers in Korea: role of fourth-generation cephalosporins. The Korean Antimicrobial Resistance Study Group. AB - Levels of resistance to the "third-generation" cephalosporins among isolates of clinical bacteria in Korea have been increasing at a rapid rate. This study evaluated the activity of cefepime, a "fourth-generation" cephalosporin, and six other broad-spectrum beta-lactam antimicrobials (cefpirome, ceftazidime, ceftriaxone, imipenem, piperacillin/tazobactam 4 micrograms/mL fixed concentration[, oxacillin) against 404 isolates of clinical bacteria from Korea. Susceptibility profiles of each isolate were established using the Etest (AB BIODISK, Solna, Sweden) method of susceptibility testing. Only the carbapenem imipenem was > 90% effective in inhibiting each of the species tested (Escherichia coli, Klebsiella, spp., Citrobacter spp., Enterobacter spp., indole positive Proteae, Serratia spp., Acinetobacter spp., Pseudomonas aeruginosa, and oxacillin-susceptible staphylococci). Imipenem was followed by cefepime > cefpirome > piperacillin/tazobactam > ceftazidime > ceftriaxone in overall rank order of usable spectrum against the isolates tested. Extended spectrum beta lactamase producing phenotypes were much more prevalent among the Klebsiella spp. (48.8%) than the E. coli (5.0%) isolates. Cefepime was much more active than cefpirome, 95.1% susceptible as compared with 70.7% susceptible, against the 41 isolates of Klebsiella spp. The results of this study corroborates findings from earlier studies with levels of resistance to the broad-spectrum beta-lactams in Korea continuing to rise indicating the need for intervention strategies. PMID- 10668592 TI - In vitro evaluation of cefepime and other broad-spectrum beta-lactams in eight medical centers in Thailand. The Thailand Antimicrobial Resistance Study Group. AB - The introduction of cephalosporins has had an important impact on the resistance rates to several clinically utilized beta-lactam antimicrobial agents. Most Thailand medical centers have not documented the levels of emerging resistant pathogens causing invasive infections. This study shows using reference-quality MIC techniques (Etest, AB BIODISK, Solna, Sweden), that carbapenem), "fourth generation" cephalosporins (cefepime and cefpirome), and piperacillin/tazobactam were the most active agents tested against Gram-negative bacilli (Escherichia coli, Klebsiella spp., Enterobacter spp., Citrobacter spp., Serratia spp., indole positive Proteae, Acinetobacter spp., Pseudomonas aeruginosa, and oxacillin susceptible Staphylococcus spp. when compared to "third-generation" cephalosporins (ceftazidime and ceftriaxone). The rank order of activity for all species was imipenem (2.9% resistant) > cefepime (7.7%) > piperacillin/tazobactam (11.1%) > cefpirome (13.4%) > ceftriaxone (21.1%) > ceftazidime (29.9%). The incidence of extended spectrum beta-lactamase production among E. coli (15.7%) and K. pneumoniae (45.6%) was significant. Cefepime and imipenem were active against the majority of these isolates. The activity of cefepime was also shown to be very good against, 1) organisms capable of producing AmpC enzymes, 2) staphylococci species that were susceptible to oxacillin, and 3) many strains of nonfermentative Gram-negative bacilli. The prevalence of antimicrobial resistance in Thailand seems to be quite high among certain commonly encountered pathogens, and imipenem and cefepime have activity (susceptible and intermediate potency) against > 90% of these organisms. PMID- 10668593 TI - Summation: beta-lactam resistance surveillance in the Asia-Western Pacific region. PMID- 10668594 TI - [First clinical episode of bipolar disorders: a study within a population of bipolar I and bipolar II French patients]. AB - Clinical symptoms of bipolar disorders onset act as a prognostic risk-factor. Discrepancies of data are related with geographical or cultural conditions. Within a patient population of bipolar (ICD 10) in and out patients of a psychiatric department, manic or hypomanic disorders initiate the space disease in 33% of the cases theses features are similar within the western psychiatric population. In a maghrebian population this proportion reaches 50%. A percentage of 65% of bipolar 1 patients was found within our sample. Sex ratio is 1 for bipolar 1, when, for bipolar 2 disorders sex-ratio was superior to 1, in favor of females. Mean age of the first episode of the disease was younger for patients with a familial history of the disease. PMID- 10668595 TI - [A study of endogenous evoked potentials (P300 and CNV) in retired patients]. AB - Endogenous evoked potentials of two types (CNV and P300 complexes) as well as their respective reaction times were recorded in 29 elderly, active subjects (15 men and 14 women) between 60 and 84 years of age. Our subjects also passed anxiety, depression and cognitive scales. The different results were then analyzed in relation with each other and in relation with age and sex. No age related differences were found in the principal elements measured in the P300 complex, but several elements of the CNV complex were clearly shown to be age sensitive. Sex-related differences were found in the N2 and P3a potentials. CNV morphology (Tecce type A) and cognitive scores were found to be related to Spielberger's anxiety-trait sub-scale. Several results also highlighted the heterogeneity of this age group. The need for complementary studies in this age group is then discussed with the hope that neurophysiology can become a useful tool in psychogeriatrics. PMID- 10668596 TI - [Stressful life events: models and methodology. Critical considerations]. AB - Many recent studies tend to show that life events play an important role in provoking, solving or maintaining psychic troubles. The proposed approach is interesting as far as it brings back into the model many psychosocial factors implied in psychic or somatic problems. We wanted to review the interest of the main theoretical and methodological approaches in that field and also to stress their drawbacks. One many improve the life events approach by taking into account the difficulties associated with that type of research and by avoiding an oversimplification of the relationship between life events and health. The links are complex, the impact of life events results from depressive mechanisms, adjustment strategies, social support, age, sex, personality, history, past experience and sociocultural representations. This complex picture compels clinicians to take a complementarist attitude when all the theories are scrutinized in order to understand the links between life events and health. From a methodological point of view, the most relevant approach seems to be the combination of standardized scales and semi-structured interviews allowing a contextual approach of events. PMID- 10668597 TI - [Heroin abuse, autobiographical memory and depression]. AB - The early psychiatric interviews with opiate addicts are characterized by three features: 1) the patient has a very factual and objective conversation, 2) the evaluation of the autobiographical memory is very difficult, 3) there is a high prevalence of affective disorders responsible for an impairment in cognitive functions. Therefore we have two aims: First, to compare episodic and semantic autobiographical memory in opiate addicts and healthy controls. Autobiographical memory is the knowledge a person has about oneself and his past. Personal semantic memory is the knowledge of the biographical facts, general knowledge and beliefs about oneself. Autobiographical episodic memory concerns recollections of personal events clearly delineated in time and space. Second, to estimate the impact of depression on the ability to produce autobiographical recollection in a population of opiatre addicts. Participants were consecutive attenders of a methadone outpatient clinic who are multiple drug dependent patients consuming mainly heroine. The first investigation took place in entry and after two months. We have recruited 21 patients with a mean duration of intoxication of 11 years. Ten of these patients have been investigated again after 2 months and 8 of them have been included in a methadone maintenance program. The patients'investigation comprised two parts: first, the evaluation of autobiographical memory (only assessed at entry of the study) with an autobiographical fluency test and the semi-structured autobiographical memory interview of Kopelman; second, the psychiatric assessment included self-rating questionnaires and observer-rating questionnaires. Opiate addicts showed a decrease in episodic autobiographical memory but an increase in semantic affective memory and objective modalization. In the fluency test, there was no difference in the number of evoked items between opiate addicts and healthy controls. The educational level influences several results. The possible explanations of these results are the action of the toxic products and a particular psychic functioning. The lack of correlation between autobiographical memory and affective disorder suggests the implication of the drugs in the emergence of memory deficits. The improvement of depressive symptomatology after two months occurring without psychotropic drugs suggests the transient feature of depression and emphasises on non-pharmacological aspects of treatment. PMID- 10668598 TI - [The study of the impact of the consensus conference "Strategies for long-term therapy of patients with schizophrenia"]. AB - The case management, treatment and psychosocial rehabilitation of schizophrenic patients is an important part of the activity of the psychiatric sector and takes up many human, scientific, organizational and financial resources. The best way to reach satisfactory results for the individual patient is still uncertain and current practice in France shows noticeable variations that have been rarely investigated in terms of outcome. A consensus conference (CC) on "Strategies for long-term therapy of patients with schizophrenia" was therefore held in Paris in 1994 to produce accurate guidelines designed to help both clinicians and patients and to improve practice. It was organized by the French Federation of Psychiatry, the National Union of Friends and Relatives of Mental Patients, and the National Agency for the Development of Health Evaluation. The conclusions of the CC were mailed, in the form of a booklet, to members of these associations (psychiatrists and relatives) and were reported in the medical and general press. METHODS: The impact of the CC was judged by (a) the psychiatrists'awareness of the existence of the CC, (b) their knowledge of its conclusions, and (c) changes in practice. The following were analyzed: press coverage; requests for the booklet; the results of a survey of a representative sample of 396 psychiatrists two years after the CC; prescription changes in the public sector in a cohort of 2,407 schizophrenic patients under treatment at the time of the CC; prescriptions to psychotic patients by a representative sample of psychiatrists in private practice. RESULTS: Awareness: Articles on the CC were published in 27 journals and newspapers, 30,000 booklets were distributed and 8,348 were mailed in response to 1,121 spontaneous requests; 78% of the psychiatrists interviewed said they were aware of the existence of the CC and 70% said they were aware of the conclusions. Knowledge: The psychiatrists' declared practice conformed with CC conclusions 41%-85% of the time depending on the recommendation. No difference in practice was noted between the psychiatrists who said they knew of the recommendations and those who said they did not. Changes in practice: A significant but small improvement in prescription habits was noted for a principal recommendation ("just one neuroleptic is enough"). One-neuroleptic prescriptions increased from 51.1% the year before the CC to 56.4% two years after the CC. The increase mainly concerned the most recently treated patients. However, during the same time-span, prescriptions of anti-cholinergics plus neuroleptics rose from 48.2% to 54.3%. CONCLUSION: It is difficult to attribute changes in practice to a CC. However, the impact of the CC seemed real even if inconstant and not great enough. Clearly, to enhance impact an action plan is needed. It should include corrective measures and focus on additional dissemination efforts, teaching and training programs, and updating of guidelines if necessary. PMID- 10668599 TI - [Addiction and personality traits: sensation seeking, anhedonia, impulsivity]. AB - This study presents the evaluation of three dimensional traits of personality (Sensation Seeking, Anhedonia, Impulsivity) among 65 patients admitted in a psychiatric ward, with or without addictive behaviors. Our objective is to establish that these personality traits are commun to all addictive behaviors and to test the hypothesis that high scores on the three scales are linked to a greater probability of presenting with addictive behaviors. The two most frequent types of addiction were alcoholism and drug abuse. The subjects presenting with one or several addictive behaviors had higher average scores on the three scales. Our results printed in the same direction for the subjects having shown an addictive behavior in their past history. The risk to present with an addictive behavior increased with the total scores of these self-report questionnaires. There was a significant relationship between 3 sub-dimensions on the Sensation Seeking Scale and addictive behavior. Each time sub-scores of boredom susceptibility, disinhibition and thrill and adventure rise by one, the risk to present with an addictive behavior is multiplied by 1.4 for the first two and by 1.3 for the third one. Subjects with high scores on the anhedonia and impulsivity scales respectively show a risk multiplied by 1.6 and 3.3 of developing an addictive behavior. These results of this transverse study confirm the link between addiction behavior and these three personality traits. PMID- 10668600 TI - [Selective attention and schizophrenia before the administration of neuroleptics]. AB - In recent years, the presence of attention deficits has been recognized as a key feature of schizophrenia. Past studies reveal that selective attention, or the ability to select relevant information while ignoring simultaneously irrelevant information, is disturbed in schizophrenic patients. According to Treisman feature-integration theory of selective attention, visual search for conjunctive targets (e.g., shape and color) requires controlled processes, that necessitate attention and operate in a serial manner. Reaction times (RTs) are therefore function of the number of stimuli in the display. When subjects are asked to detect the presence or absence of a target in an array of a variable number of stimuli, different performance patterns are expected for positive (present target) and negative trials (absent target). For positive trials, a self terminating search is triggered, that is, the search is ended when the target is encountered. For negative trials, an exhaustive search strategy is displayed, where each stimulus is examined before the search can end; the RT slope pattern is thus double that of the positive trials. To assess the integrity of these processes, thirteen drug naive schizophrenic patients were compared to twenty normal control subjects. Neuroleptic naive patients were chosen as subjects to avoid the potential influence of medication and chronicity-related factors on performance. The subjects had to specify as fast as possible the presence or absence of the target in an array of a variable number of stimuli presented in a circular display, and comprising or not the target. Results showed that the patients can use self-terminating search strategies as well as normal control subjects. However, their ability to trigger exhaustive search strategies is impaired. Not only were patients slower than controls, but their pattern of RT results was different. These results argue in favor of an early impairment in selective attention capacities in schizophrenia, which appears before the introduction of neuroleptics. The attention performance was also shown to present some association to clinical symptoms. PMID- 10668601 TI - [Associations and interactions: tricyclic antidepressants and selective serotonin reuptake inhibitors]. AB - Following the commercialization of the SSRIs clinicians described cases of drug interactions with tricyclic antidepressants among their patients. When combining tricyclic antidepressants and SSRIs clinical side effects and elevated plasma levels of tricyclics appeared. A better knowledge of the cytochrome P450 system allows to understand the mechanism of such drug interactions. The cytochrome P450 is composed of a group of isoenzymes, which are classified, into families and subfamilies on the basis of amino acid sequence homology. A number of the cytochrome genes have a genetic polymorphism responsible for poor and extensive metabolisers. The clinical importance of genetic polymorphism is highly dependent upon the therapeutic index. Thus, poor metabolisers, will experience side effects and rapid metabolisers prone to therapeutic failure. Concerning pharmacological issues SSRIs have a great affinity for at least one of the isoenzymes which accounts for drug interactions. Due to the inhibitory potential of the SSRIs drug interactions occur with tricyclics that have a narrow therapeutic index. The SSRIs do not exert the same inhibitory effect on the various isoenzymes. The inhibitory activity for an isoenzyme depends on the molecule of the SSRIs. In clinical practice, the associations between tricyclics and SSRIs should be practiced with caution. It is recommended to decrease the tricyclic dose before administering the SSRI, to start with low doses of the SSRI and to take into account the therapeutic index. Although the coadministration of tricyclics and SSRIs can produce adverse reactions it has also two main interests in clinical practice. First, the drug combination enhances clinical response to treatment. Secondly, it converts non-responders of pharmacological treatment to responders. Used with caution the association of tricyclics and SSRIs is well tolerated. However, it should be kept in mind that a single drug therapy should be tried first. These data show the complexity of drug interactions. PMID- 10668602 TI - [Case report: electroconvulsive therapy during pregnancy]. AB - The treatment for psychiatric disorders in pregnancy remains difficult to implement. We report the case of a 28-year-old woman, 20 weeks pregnant when admitted in our psychiatric department. She presented severe depressive disorder, associated with agitation, and psychotic symptoms as delusion and hallucinations occurred. The patient had a history of recurrent mood disorders dating back to eight years before the current admission, including some atypical episodes (psychotic symptoms only), and alternating with free periods without any trouble. A non-specific personality disorder is also probably present. We first used antidepressant (clomipramine) and sedative phenothiazine drugs. Because of the lack of therapeutic efficacy, three weeks later we tried another pharmacologic prescription, that also failed to improve the patient' status. It was hence decided to proceed with electroconvulsive therapy. We describe here the management of the courses, especially the careful monitoring and the anesthetic features we employed, among which endotracheal intubation, oxygen supply, real time ultrasonography, and recording uterine contractions and fetal heart rate. All theses measures were applied within a surgical-obstetrical theatre. Nine bifrontal courses were performed in five weeks. They rapidly and completely improved the psychiatric symptoms. No sign of fetal neither maternal bad tolerance occurred. While the patient had been authorized to leave hospital, in 34th weeks amenorrhea a routine ultrasonographic examination discovered worrying fetal ascites signs. After the emergency caesarean delivery, the male newborn child undergone immediately surgical treatment for vascular meconium peritonitis, but died nine days later with metabolic post-surgical troubles. This fatal outcome after electroconvulsive therapy leads us to discuss its possible involvement, and in a more general way the safety and place of this treatment in pregnancy psychiatric disorders. They remain critical situations in which therapeutic methods should be rapidly decided. The authors wish others practitioners to bring new case-reports in order to assess the ECT safety-use during pregnancy. PMID- 10668603 TI - [Neuropsychiatric disorders induced by MDMA ("Ecstasy")]. AB - If neurotoxicity of MDMA (ecstasy) is now well documented in animals, it is not the same in humans. MDMA intoxication puts the problem of its possible link with the serotonin syndrome and the neuroleptic malignant syndrome. Neuropathological consequences following MDMA intake have been reported, including hemorrhaging and cerebral infarction, cerebral venous sinus thrombosis, and acute inflammatory CNS disease. However, the physiopathology of these complications remains unclear. In the same way, there have been various reports that have attributed MDMA to precipitating the onset of a wide range of psychiatric disorders including sleep disorders, cognitive disorders, panic attacks, depression, flashbacks, psychosis and severe paranoia. Findings suggest that these psychiatric manifestations might be consequences of MDMA induced brain serotonin neurotoxic lesions. All these data are examined from a critical review of the literature. PMID- 10668604 TI - [Survey on the announcement of schizophrenia diagnosis in France]. AB - Medical information for the general public, patients and their families is a current Public Health priority. What information can be given to a patient suffering from schizophrenia, whose understanding and judgement capacities are supposedly affected by this mental disease? In the United States, 70% of psychiatrists inform patients of schizophrenia and diagnosis of schizophreniform disorder, while in Japan less than 30% do this. The lack of information given to the general public on the disease may contribute to reinforcing the difficulty in announcing the diagnosis. Indeed, the beliefs and attitudes of the patient, his/her family, the general population and health carers concerning the disease do not match up. However, the first two years seem to be a main issue for the subsequent evolution of the disease. No specific data on the attitude of French clinicians with respect to the announcement of the diagnosis is available. In the current legal context and in view of the advances in treatment, we have carried out a survey among French psychiatrists. It is an auto-questionnaire, transversal epidemiological, descriptive and analytical. The questionnaire was sent to a population of 12,958 psychiatrists. It comprised 48 questions: 7 referred to the socio-demographic and professional characteristics of the subjects, 22 to the attitude with respect to the announcement of the diagnosis to the patients, and the last 18 concerned the attitude with respect to the announcement of the diagnosis to the families. 1,691 questionnaires were returned by free post and analysed. The socio-demographic characteristics of the sample are close to those of French psychiatrists as a whole. The number of patients suffering from schizophrenia in the active files of the psychiatrists is 24% (+/- 21.4) on the entire sample. Approximately a third (37.8%) of psychiatrists deem it necessary to announce the schizophrenia diagnosis and approximately two thirds (69.5%) declare that they sometimes announce it. Among the patients suffering from schizophrenia in the active files of the psychiatrists who responded, approximately a third (34%) were informed of their diagnosis. The main reasons for not announcing the diagnosis are firstly the "reticence to give a diagnosis label" and secondly "the functional incapacity of the patient to understand the concept". The alternative diagnosis term most commonly used is "psychosis" (46.5%). However, 48.1% of practitioners state that the announcement of a specific diagnosis allows a better therapeutic combination. Depending on the proportion of patients suffering from schizophrenia in their active file presented in two categories (< 10% and > 10%), psychiatrists significantly most frequently announce the specific diagnosis (17.3% vs 25.3%, p < 10(-3). A statistically significant proportion of younger psychiatrists (44.4 vs 46.3, p < 10(-3) with fewer years of practice (14.1 vs 15.8), more often believe that it is necessary to announce the diagnosis. The rate of response (13.5%) for this type of survey seems high, which could indicate a high interest among psychiatrists with respect to this question. Our data showed the existence of a correlation between age, number of years in practice, type of practice and the proportion of patients suffering from schizophrenia in the active file on the one hand and the attitude of the psychiatrists with respect to the announcement of the diagnosis on the other hand. It is possible that the multi-disciplinary team work of public practice psychiatrists and the fact that they are more often confronted with schizophrenic disease facilitate the announcement of this diagnosis. In the survey population, the inability to give a diagnosis may be related to the questions of the practitioners about the capacity of the subjects to understand, the lack of precision of this diagnosis, the fear of disheartening the patients and the absence of curative treatment. The risk of suicide does not seem to be one PMID- 10668605 TI - [Diagnostic criteria for fronto-temporal lobe degeneration]. AB - Circumscribed atrophy of the frontal and temporal lobes (frontotemporal lobar degeneration) accounts for about one fifth of cases of primary degenerative dementia occurring before the age of 65. It produces three prototypical clinical syndromes. The most common is frontotemporal dementia, characterized by personality change and profound alteration in social conduct and associated with bilateral atrophy of the frontal and anterior temporal lobes. Progressive non fluent aphasia is characterized by difficulty in verbal expression, anomia and phonemic errors in the presence of relative preservation of comprehension and associated with atrophy predominantly of the left hemisphere. In semantic dementia there is fluent speech with semantic errors and severely impaired comprehension and naming, together with a visual associative agnosia, resulting from bilateral atrophy of the inferior and middle temporal gyri. The clinical syndromes occur with either of two main histological types: prominent microvacuolar change, without specific histological features (frontal lobe degeneration-type), severe astrocytic gliosis with or without ballooned cells and inclusion bodies (Pick-type). To improve clinical recognition and advance understanding of this relatively common form of cerebral degeneration, members of an international workshop on Frontotemporal Lobar Degeneration developed consensus criteria, building upon earlier published clinical diagnostic guidelines for frontotemporal dementia. The consensus criteria reported here specify core and supportive features for each of the prototypical clinical syndromes: frontotemporal dementia, progressive aphasia and semantic dementia, as well as providing broad inclusion and exclusion criteria for the generic entity of frontotemporal lobar degeneration. PMID- 10668606 TI - [Construction and validation of visual analogue scale for risk assessment]. AB - This paper describes the factor structure and validity of a visual analogue scale designed to rate risk proneness: EVAR (EVAluation of Risk). Risk behaviors and sensation seeking are related, therefore we correlated EVAR (24 100 mm items) with Zuckerman Sensation Seeking Scale (SSS) (40 conventional items) in a population of 199 volunteers. Subjects had an age range of 18-68 years (mean age of 31), and 49 were females. Factor analysis using a principal component solution and an orthogonal rotation of the factor matrix were computed. Five factors were selected, accounting for 46 per cent of the total variation: F1 "self control", F2 "danger seeking", F3 "energy", F4 "impulsiveness", F5 "invincibility". F1, F2 and F3 from EVAR were correlated with SSS factors "general", "danger seeking", "experience seeking" and "boredom susceptibility". F4 was correlated with "general", "boredom susceptibility" and "disinhibition" whereas F5 was not correlated with any of SSS factors. This visual analogue scale is likely to reveal change in subjects, regarding risk proneness, when submitted to various stressors (sleep deprivation, fatigue...). PMID- 10668607 TI - [Methodological proposals for the evaluation of the mood stabilizing effect of an antipsychotic drug]. AB - Antipsychotic drugs are widely used in the long term treatment of bipolar and schizoaffective disorders. Nevertheless clinical trials devoted to the specific assessment of the mood stabilizing effect of these drugs are quite rare. Among recent studies only those by S. McElroy on olanzapine and by R. Bowden on divalproex have included mood symptoms as inclusion and outcome criteria. This paper reviews the different methodological proposals both in term of protocol designs (retrospective, naturalistic, longitudinal and prospective studies) and of efficacy criteria (time to relapse, reasons for drop outs, treatment switch, functional status). PMID- 10668608 TI - [Orbitofrontal syndrome in psychiatry]. AB - Orbitofrontal syndrome is a variant of frontal lobe syndrome in which behavioural disturbances are prevailing. It results from bilateral lesions of the orbitofrontal cortex and the medial face of frontal lobe. Patients present disorganized hyperactivity. They are distractable, impulsive, euphoric and unable to abide by social rules. They often have instinctive disinhibition (hypersexuality, hyperphagia and urinary behaviour disorders). In spite of severe behavioural disturbances cognitive functions are often intact so that orbitofrontal syndrome may be confounded with two psychiatric disorders: mania (or hypomania) and antisocial personality disorder. In this article we present a case report of orbitofrontal syndrome which was initially misdiagnosed as mania. Clinical features and possible modes of presentation of this syndrome are discussed. It is suggested that serotonin reuptake inhibitors may be of some use in this disorder. PMID- 10668609 TI - [The concept of supersensitivity psychosis. The particular case of clozapine]. AB - Neuroleptics are the main biological treatment for psychotic patients. The brutal withdrawal of a neuroleptic treatment may induce an important aggravation of the psychotic symptoms. A few of those relapses may occur very early after the interruption of treatment; they are often associated with a modification of the symptoms and an unfavorable evolution in the course of the illness. Using those clinical observations a few authors have developed the concept of supersensitivity psychosis to explain those kinds of relapses and to formulate hypothesis about tolerance and resistance to neuroleptics. They focus on the possible correlation between supersensitivity psychosis and tardive dyskinesia. We report three cases of a dramatic aggravation of the psychotic symptomatology following the withdrawal of clozapine in three schizophrenic patients resistant to classical neuroleptic treatment. According to the clinical data and to the physiopathological hypothesis, the concept of supersensitivity psychosis can have implications in the therapeutic management of resistant schizophrenic patients. PMID- 10668610 TI - [Predictive factors for patient maintenance on buprenorphine high dosage treatment: a naturalistic study in primary care]. AB - High dosage buprenorphine is actually the principal treatment for substitution medication in France. Clinical trials have demonstrated the clinical efficacy of HD buprenorphine for narcotic addiction, but few data are published concerning the prognostic factors of treatment response in daily practice. A naturalistic study was performed in 1998. 200 generalist practitioners were recruited and 956 patients were included. Sociodemographic, medical and addiction history were collected. A quantitative socio-comportemental and medical indicator (SCMI) was performed. The psychometric properties of the SCMI were analyzed. Simple and multivariate analysis was performed. Patients with good social adjustment and past withdrawal are good responders to HD buprenorphine. Not treated psychiatric pathology was a prognostic factor associated with a relatively poor response to HD buprenorphine. A long duration of treatment (one year) and a clear therapeutic program were associated with good response. PMID- 10668611 TI - [Abuse of and dependence on zolpidem: a report of seven cases]. AB - Zolpidem is an hypnotic drug that belongs to the imidazopyridine family. Its chemical structure is different from that of benzodiazepines though both type of drugs bind specifically to the same site of the GABA-A macromolecular complex: the omega 1 benzodiazepine receptor. This mechanism of action could be responsible for the predominantly hypnotic properties of zolpidem and its reduced liability to induce dependence in comparison with benzodiazepines. Yet, several cases of zolpidem abuse and dependence have been published recently. We report seven cases, from which three are detailed, of zolpidem abuse and/or dependence. These patients did not suffer from sedative effects of this drug despite important doses. We even noticed stimulating and euphorising effects in two of these patients, an effect that may explain at least in part the dependence to zolpidem. We will discuss the clinical similarities existing between zolpidem and benzodiazepines' effects. Furthermore we will discuss a molecular genetic hypothesis that may explain the differential effect of a specific benzodiazepine ligand on its receptors. PMID- 10668612 TI - [New antipsychotics in the treatment of schizophrenia. A European survey]. AB - Atypical antipsychotics (clozapine, risperidone, olanzapine, quetiapine, sertindole) make up a much larger proportion of the prescriptions for antipsychotic medications in the United States than in Europe. It is certain that these atypical neuroleptics are not all available throughout Europe; nonetheless the size of the disparity reveals certain tendencies in the different nations. In an attempt to identify the reasons for the lesser usage of the new antipsychotics in Europe, a telephone survey was conducted with 686 psychiatrists in 9 countries. This opinion survey was intended to identify, using open-ended questions and multiple choice, the reasons for which practitioners have or have not used the new antipsychotics; their perceived advantages and disadvantages in comparison with typical antipsychotics; and the hindrances in prescribing them. The results revealed that the new antipsychotics have a positive image with psychiatrists: whereas they estimate the proportion of their patients using the new antipsychotics to be at 50% (an amount larger than the objective European amounts), more than 80% of psychiatrists say they would be ready to use them more frequently if certain problems were overcome. Significant obstacles related to the product are the cost and the lack of a depot formulation; two hindrances with respect to the patient are the difficulty in using them in an emergency situation, and the fear of destabilizing a patient who is well-controlled with a classical treatment. The discussion re-addresses these points, using literature relevant to the products and the patients. The European data, which are often homogeneous, are discussed as a whole, with the exception of certain characteristics that are specific to an individual country. French psychiatrists, notably, serve as an exception, because they describe themselves as being more restrained in their prescriptions because of the lack of a depot formulation rather than because of the cost of the product. PMID- 10668613 TI - [Between compliance and freedom: the patient's statement]. AB - According to the patient, obtaining a good compliance is related to a good relationship with his practitioner; this relationship is directly connected to being a good listener for the patient, and not only for their symptoms. If the individual motivation is very important at the beginning, it will be itself greatly influenced by the relationship between the practitioner and the patient. It is one of the rare factors with a positive correlation with compliance, that's what almost all of the researches on medical psychology have observed. Once the problem is sumed up, it's the turn to speak for the patient, who explains how he considers compliance. PMID- 10668616 TI - [Drug-induced psychosis versus lupus-related psychosis: a report of a case]. PMID- 10668615 TI - [About ethics in psychiatry and psychiatrists facing ethics]. AB - The plug in account of the suffering, notably psychological, in a consultation, puts the problem of the relationship between suffering and ethics. However, the originality of the ethical step is justly not to be confined to the social norm conformism, but being specific to the individual dimension. The psychiatric pathology offers in this area of particularities interesting. The neurotic, as the obsessed, suffering inwardly pathological manifestations that he judges absurd, replies to the medical moral in asking a care. The psychotic, which projects his suffering on the other, does not feel sick, requests no therapeutic assistance. As such he contests the medical order in an immoral position by definition, and the patient represents from then on a social and medical scandal. In front of a such clinical diversity, we can easily underline that approaching the theme of ethics in psychiatry isn't a well-off exercise, and necessitates a precise locating registered in the history of the patient. PMID- 10668614 TI - [Abuse of tianeptine. A case report]. AB - The authors report a case of tianeptine abuse in a 30 year-old woman. After a medical prescription of the recommended dosage of 12.5 mg 3 times daily of oral tianeptine for a depressive illness, the patient spontaneously increased the dosage which after two months reached 150 tablets per day. No severe toxic effects were observed. As adverse effects, the patient, in the beginning of this high treatment period suffered from nausea, vomiting, abdominal pain, anorexia with weight loss, constipation. These side effects progressively disappeared. The biological tolerance was excellent, and hepatic parameters were not affected. The patient experienced and seek a psychostimulant effect. After seven months of such a therapy, she was hospitalized to undergo a withdrawal. The discontinuation of the tianeptine treatment occurs in four days. A withdrawal syndrome marked by myalgia, and cold feeling was transient, and alleviated by sedative phenothiazine (cyamemazine) and myorelaxant benzodiazepine (tetrazepam). PMID- 10668617 TI - [Perinatal psychopathology: some theoretical considerations]. PMID- 10668618 TI - In memoriam: Mara Selvini-Palazzoli, M.D. (1916-1999). PMID- 10668619 TI - Dialectical behavior therapy--family skills training. AB - Over the past three decades, family interventions have become important components of treatment for a number of psychiatric disorders. To date, however, there has been no family treatment designed specifically for borderline personality disorder patients and their relatives. This article describes one short-term family intervention called Dialectical Behavior Therapy-Family Skills Training. Based on Linehan's Dialectical Behavior Therapy (DBT), borderline patients' behavioral patterns are thought to result from a lifelong transaction between emotional vulnerability and invalidating features of the social and familial environment. Individual DBT focuses on reducing individual emotion dysregulation and vulnerability and enhancing individual stability. The complementary family interventions proposed in this article aim to: 1) provide all family members an understanding of borderline behavioral patterns in a clear, nonjudgmental way; 2) enhance the contributions of all family members to a mutually validating environment; and 3) address all family members' emotion regulation and interpersonal skills deficits. PMID- 10668620 TI - Reconstructing the Brothers Grimm: new tales for step-family life. AB - Step-families are situated within the sociopolitical context of family change and are examined as a prototype of the "post-modern" family. This essay looks at the cultural construction of step-family life and proposes a model for collaboratively reconstructing stories that liberate step-relationships from the legacy of the Brothers Grimm, deconstructing the stories of failure, insufficiency, and neglect. Building on narrative and social constructionist ways of thinking about families, the concept of side-shadowing (a hermeneutic approach from literary/historical criticism) is introduced to elucidate how therapists can help family members discover ways of thinking, feeling, and behaving that are both more personally satisfying and more congruent with the changed context of family life. Two therapeutic challenges are high-lighted: reconceptualizing what it means to be a step-family and coming to terms with differential attachment in relationships while working with step-families. The essay ends with a fairy tale for the 21st century. PMID- 10668621 TI - Attitudes toward and perceived psychosocial impact of female circumcision as practiced among the Bedouin-Arabs of the Negev. AB - The present pilot study examines attitudes toward and the perceived psychosocial impact of circumcision as practiced among the Bedouin-Arabs of the Negev, Israel. A convenience sample of 24 women participated in the study: 12 who had experienced the ritual, and 12 who had not, but who had witnessed or been told about the practice of the ritual on women in their extended families. Two research instruments were used: a structured questionnaire, and a semi-structured open-ended interview. Data showed differences in subject responses depending on the research tools. The structured questionnaire revealed that women who had experienced the circumcision gave legitimization and cognitive rationalization to it. In contrast, the semi-structured interview revealed that these same subjects reported insult: traumatization, direct negative influences, and narcissistic insult, and described emotional difficulties during the research interviews. The findings indicated that they had difficulties in mother-daughter relationships and trust. Implications of the ritual on the continuity of polygamy and marital/sexual problems are discussed. PMID- 10668622 TI - A culturally sensitive approach to therapy with immigrant families: the case of Jewish emigrants from the former Soviet Union. AB - This article is based on accumulated clinical experience in Israel with families that emigrated from the former Soviet Union. It describes a culturally sensitive systemic intervention with two such families: a single-parent family, and a family that exhibited physical violence. Relevant cultural characteristics of family patterns and parent-child relationships in Jewish-Soviet families are reviewed. It is demonstrated how a cross-cultural perspective may affect the interpretation of presented problems and result in a less pathological perspective. It is further illustrated how universal intervention techniques combined with culturally sensitive approaches may produce positive effects in therapy. PMID- 10668623 TI - Family interaction styles of children with depressive disorders, schizophrenia spectrum disorders, and normal controls. AB - Family interaction processes during a problem-solving task were examined in children with depressive disorders, children with schizophrenia-spectrum disorders, and a normal control group of community children screened for the absence of psychiatric disorder. Major findings were: a) children with depressive disorders were more likely than children with schizophrenia-spectrum disorders and children with no psychiatric disorder to direct guilt-inducing comments toward their parents; and b) parents of children with schizophrenia-spectrum disorders were more likely to direct harsh critical comments toward the child than were parents of depressed children or parents of normal controls. In addition, children's and mothers' use of benign criticism was linked, while children's harsh criticism was associated with intrusion from the father, and children's self-denigrating comments were related to specific paternal criticism. Implications of these results for understanding transactional processes associated with childhood-onset depressive and schizophrenia-spectrum disorders are discussed. PMID- 10668624 TI - Racial and gender differences in expressed emotion and interpersonal control in families of persons with schizophrenia. AB - Expressed Emotion (EE) has been shown to be predictive of course or severity in many illnesses, but the studies have been largely of white middle-class patients. This study examined gender and racial differences in parental EE level and communication patterns between the parent and patient with schizophrenia, using data from the NIMH Treatment Strategies in Schizophrenia study. Dialogues (n = 140) from 54 patient-parent dyads were coded into the Relational Control Coding System. Resultant data (n = 13,605 sequences) were analyzed with log-linear models. Results show that the relationship between control and EE level was stronger in African American families compared to Caucasians. Gender differences were as expected, with daughters less competitive and more deferential to their parents. Although the total number of high-EE parents with daughters was small, patterns in these families showed parents who responded submissively in contrast to the competitive symmetry in families with male patients. PMID- 10668625 TI - Factor analyses of the family assessment device. AB - The Family Assessment Device (FAD) operationalizes the McMaster Model of Family Functioning, which has been used in numerous studies, translated into seven languages, and is regarded as one of the most researched family assessment tools available. However, exploratory and confirmatory factor analyses using the 7-by-7 matrix of subscale correlations from the original validity study on the FAD (Epstein, Baldwin, & Bishop, 1983) indicated that the FAD subscales overlap substantially and do not assess unique dimensions of family functioning. Results of our study suggest that the conservatively best use of the FAD is using the General Functioning subscale as a summary score. A model that fits the data marginally better than the General Functioning score and a Measurement Error model, however, consisted of "Collaboration" and "Commitment" latent factors. These results illustrated the need for more extensive validity research on the FAD, because interpretation of the factors and subscales had to rely heavily on face validity. PMID- 10668626 TI - Studies on a "jumping gene machine": higher-order nucleoprotein complexes in Mu DNA transposition. AB - Studies in my lab have focused on DNA transposition in the bacterial virus, Mu. In vitro studies have shown that Mu DNA transposition is a three-step process involving DNA breakage, strand transfer and DNA replication. In the first step, a nick is introduced at each end of the transposon. The liberated 3'-OH groups subsequently attack a target DNA molecule resulting in strand transfer. The transposon DNA, now covalently linked to the target, is finally replicated to generate the transposition end-product, referred to as a cointegrate. The DNA cleavage and strand transfer reactions are mediated by a "jumping gene machine" or transpososomes, which we discovered in 1987. They are assembled by bringing together three different DNA regions via a process involving multiple protein-DNA and protein-protein interactions. The action of four different proteins is required in addition to protein-induced DNA bending or wrapping to overcome the intrinsic stiffness of DNA, which would ordinarily prohibit the assembly of such a structure. Transpososome assembly is a gradual process involving multiple steps with an inherent flexibility whereby alternate pathways can be used in the assembly process, biasing the reaction towards completion under different conditions. PMID- 10668627 TI - Roles of protein tyrosine phosphatases in cell migration and adhesion. AB - Signal transduction pathways are often seen as cascades of kinases, whereas phosphatases are relinquished to the housekeeping function of resetting the individual elements to a resting state. However, critical biological processes such as cellular migration require a coordinated and constant remodeling of the actin cytoskeleton as well as a rapid turnover of the cell-substratum linkages that necessitate the concomitant action of antagonistic enzymes. Tyrosine phosphorylation was long known to be involved in adhesion and de-adhesion mediated via the integrin receptors. As the roles of tyrosine kinases such as focal adhesion kinase, c-Src, and Csk in this pathway are being extensively studied, increasing evidence is emerging about the importance of protein tyrosine phosphatases (PTP). In this review we discuss examples of PTPs that were recently shown to play a role in cell adhesion and migration and their mechanism of action. PMID- 10668628 TI - Identification of the Escherichia coli enzyme I binding site in histidine containing protein, HPr, by the effects of mutagenesis. AB - The structure of the N-terminal domain of enzyme I complexed with histidine containing protein (HPr) has been described by multi-dimensional NMR. Residues in HPr involved in binding were identified by intermolecular nuclear Overhauser effects (Garrett et al. 1999). Most of these residues have been mutated, and the effect of these changes on binding has been assessed by enzyme I kinetic measurement. Changes to Thr16, Arg17, Lys24, Lys27, Ser46, Leu47, Lys49, Gln51, and Thr56 result in increases to the HPr Km of enzyme I, which would be compatible with changes in binding. Except for mutations to His15 and Arg17, very little or no change in Vmax was found. Alanine replacements for Gln21, Thr52, and Leu55 have no effect. The mutation Lys40Ala also affects HPr Km of enzyme I; residue 40 is contiguous with the enzyme I binding site in HPr and was not identified by NMR. The mutations leading to a reduction in the size of the side chain (Thr16Ala, Arg17Gly, Lys24Ala, Lys27Ala, and Lys49Gly) caused relatively large increases in Km (>5-fold) indicating these residues have more significant roles in binding to enzyme I. Acidic replacement at Ser46 caused very large increases (>100-fold), while Gln51Glu gave a 3-fold increase in Km. While these results essentially concur with the identification of residues by the NMR experiments, the apparent importance of individual residues as determined by mutation and kinetic measurement does not necessarily correspond with the number of contacts derived from observed intermolecular nuclear Overhauser effects. PMID- 10668629 TI - Inhibition of retinoic acid-inducible transcription by COUP-TFI in human salivary gland adenocarcinoma cell line HSG. AB - Human salivary gland adenocarcinoma cells (HSG) express nuclear receptors, all trans-retinoic acid (at-RA) receptors (RARs), and retinoid X/9-cis-retinoic acid (9-c-RA) receptors (RXRs). In order to investigate whether the endogenous RARs or RXRs of HSG cells can induce transcription activation, the thymidine kinase promoter (TK)-driven luciferase reporter gene containing the retinoic acid response element (RARE), of RARbeta, betaRARE2-TK-Luc, was transfected into HSG cells and ligand-dependent transcription activation was examined. Luciferase activity of cell lysate increased by the treatment with either at-RA or 9-c-RA. Co-transfection of RARalpha and (or) RXRalpha-expression plasmids with the reporter gene enhanced the luciferase activity, suggesting that endogenous RARs and RXRs work as ligand-dependent transfactors in HSG cells. Reverse transcriptase - polymerase chain reaction analysis revealed that HSG cells express chicken ovalbumin upstream promoter - transcription factor I (COUP-TFI). Co-transfection of COUP-TFI-expression plasmid suppressed the at-RA-induced transcription activation of the reporter gene. Similar results were shown using a chromatin-integrated reporter gene system, using a stably transfected beta-RARE2 TK-beta-galactosidase (beta-Gal) reporter gene. The at-RA-dependent increase in the beta-Gal expression was completely inhibited by COUP-TFI. The transfection of antisense oligonucleotide of COUP-TFI squelched the RA-dependent growth inhibition induced by RAR-RXR heterodimers. Conclusively, RARs and RXRs of HSG cells are functional and play roles as transactivators in at-RA-sensitive processes such as the proliferation or differentiation of cells. COUP-TFI very likely regulates these processes by repressing the functions of these transactivators. PMID- 10668630 TI - Cell adhesion and the actin cytoskeleton of the enveloping layer in the zebrafish embryo during epiboly. AB - As the zebrafish embryo undergoes gastrulation and epiboly, the cells of the enveloping layer (EVL) expand, covering the entire yolk cell. During the epiboly process, the EVL cells move as a coherent layer, remaining tightly attached to each other and to the underlying yolk syncytial layer (YSL). In view of the central role of the actin cytoskeleton, in both cell motility and cell-cell adhesion, we have labeled these cells in situ with fluorescent phalloidin and anti-actin antibodies. We show that, throughout their migration, the EVL cells retain a conspicuous cortical actin cytoskeletal belt coinciding with cell surface cadherins. At the margins approaching the YSL, the EVL cells extend, from their apicolateral domains, actin-rich filopodial protrusions devoid of detectable cadherin. We have studied the role of the actin cytoskeleton in the maintenance of EVL cohesion during epiboly. Cytochalasin treatment of embryos induces EVL dissociation accompanied by general detachment of the rest of the embryonic cells. In the dissociating EVL cells, the cortical actin belt undergoes fragmentation with the formation of actin aggregates; cadherins, on the other hand, remain evenly distributed at the junctional cell surface. Removal of Ca2+ by ethyleneglycolbis (amino-ethyl-ether)-tetraacetic acid (EGTA) treatment also induces cell dissociation without visible disruption of the cortical actin belt. The protein kinase inhibitor (1-isoquinolinylsulfonyl)-2-methyl-piperazine dihydrochloride (H-7), which blocks acto-myosin contractility and disrupts actin cables in cultured cells, also potentiates cytochalasin-induced dissociation and promotes the projection of numerous actin-rich lamellipodial extensions. The fact that EVL cells produce microspike-like structures towards the YSL and are capable of lamellipodial activity lend further support to the suggestion (R.W. Keller and J.P. Trinkaus. 1987. Dev. Biol. 120: 12-24) that the EVL cells are not passively mobilized on the expanding YSL but actively participate in epiboly. PMID- 10668631 TI - Anion binding characteristics of the band 3 / 4,4-dibenzamidostilbene-2,2 disulfonate binary complex: evidence for both steric and allosteric interactions. AB - A novel kinetic approach was used to measure monovalent anion binding to better define the mechanistic basis for competition between stilbenedisulfonates and transportable anions on band 3. An anion-induced acceleration in the release of 4,4'-dibenzamidostilbene-2,2'-disulfonate (DBDS) from its complex with band 3 was measured using monovalent anions of various size and relative affinity for the transport site. The K1/2 values for anion binding were determined and correlated with transport site affinity constants obtained from the literature and the dehydrated radius of each anion. The results show that anions with ionic radii of 120-200 pm fall on a well-defined correlation line where the ranking of the K1/2 values matched the ranking of the transport site affinity constants (thiocyanate < nitrate approximately bromide < chloride < fluoride). The K1/2 values for the anions on this line were about 4-fold larger than expected for anion binding to inhibitor-free band 3. Such a lowered affinity can be explained in terms of allosteric site-site interactions, since the K1/2 values decreased with increasing anionic size. In contrast, iodide, with an ionic radius of about 212 pm, had a 10-fold lower affinity than predicted by the correlation line established by the smaller monovalent anions. These results indicate that smaller monovalent anions have unobstructed access to the transport site within the band 3 / DBDS binary complex, while iodide experiences significant steric hindrance when binding. The observation of steric hindrance in iodide binding to the band 3 / DBDS binary complex, but not in the binding of smaller monovalent anions, suggests that the stilbenedisulfonate binding site is located at the outer surface of an access channel leading to the transport site. PMID- 10668632 TI - Identification of sds21 in fission yeast in an inhibitor-resistant high molecular mass protein phosphatase-1 complex. AB - While characterizing the type-1 protein phosphatases sds21 and dis2 in fission yeast (Schizosaccharomyces pombe) a novel high molecular mass protein was identified with serine/threonine phosphatase activity (referred to as PP-R) that was resistant to a panel of characteristic inhibitors of protein phosphatases. Purification of the native sds21 catalytic isoform of protein phosphatase-1 (PP 1) from an S. pombe knockout strain lacking dis2 (deltadis2) resulted predominantly in identification of PP-R. To test the hypothesis that the catalytic activity of PP-R comprised sds21, a parallel purification was performed of PP-1 activity from an S. pombe knockout strain lacking sds21 (deltasds21). Both deltasds21 and deltadis2 strains exhibited similar protein phosphatase activity profiles as determined by DEAE-sepharose, Mono-Q and Superdex gel filtration chromatography. However, the peak of protein phosphatase activity from deltasds21 S. pombe that co-migrated with PP-R from deltadis2 S. pombe exhibited the sensitivity to a panel of inhibitors that was characteristic of a type-1 protein phosphatase. These data suggest that the catalytic subunit of PP-R comprises sds21 and that the resistance to inhibitors may originate from structural differences between dis2 and sds21 isoforms. A key structural feature present in sds21, but lacking in dis2, is a classical phosphorylation consensus sequence surrounding serine-145 of sds21. The previous hypothesis was that PP-1 activity among several lower eukaryotes may be regulated directly by cAMP dependent protein kinase (PKA) phosphorylation. However, this study demonstrated that recombinant sds21 is not a target for PKA in vitro. The constrained configuration of the putative PKA site on the PP-1 holoenzyme may restrict its ability to be targeted by PKA. PMID- 10668633 TI - Ischemia induced activation of heat shock protein 27 kinases and casein kinase 2 in the preconditioned rabbit heart. AB - Protein kinase C (PKC), p38 MAP kinase, and mitogen-activated protein kinase activated kinases 2 and 3 (MAPKAPK2 and MAPKAPK3) have been implicated in ischemic preconditioning (PC) of the heart to reduce damage following a myocardial infarct. This study examined whether extracellular signal-regulated kinase (Erk) 1, p70 ribosomal S6 kinase (p70 S6K), casein kinase 2 (CK2), and other hsp27 kinases are also activated by PC, and if they are required for protection in rabbit hearts. CK2 and hsp27 kinase activities declined during global ischemia in control hearts, whereas PC with 5 min ischemia and 10 min reperfusion increased their activities during global ischemia. Resource Q chromatography resolved two distinct peaks of hsp27 phosphotransferase activities; the first peak (at 0.36 M NaCl) appeared to correspond to the 55-kDa MAPKAPK2. Erk1 activity was elevated in both control and PC hearts after post ischemic reperfusion, but no change was observed in p70 S6K activity. Infarct size (measured by triphenyltetrazolium staining) in isolated rabbit hearts subjected to 30 min regional ischemia and 2 h reperfusion was 31.0+/-2.6% of the risk zone in controls and was 10.3+/-2.2% in PC hearts (p<0.001). Neither the CK2 inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) nor the Mek1/2 inhibitor PD98059 infused during ischemia blocked protection by PC. The activation of CK2 and Erk1 in ischemic preconditioned hearts appear to be epiphenomena and not required for the reduction of infarction from myocardial ischemia. PMID- 10668634 TI - Fibroblast growth factor-2 stimulates phospholipase Cbeta in adult cardiomyocytes. AB - Although fibroblast growth factor-2 (FGF-2) plays an important role in cardioprotection and growth, little is known about the signals triggered by it in the adult heart. We therefore examined FGF-2-induced effects on phosphoinositide specific phospholipase C (PI-PLC) isozymes, which produce second messengers linked to the inotropic and hypertrophic response of the myocardium. FGF-2, administered by retrograde perfusion to the isolated heart, induced an increase in inositol-1,4,5-trisphosphate levels in the cytosol, as well as an increase in total PI-PLC activity associated with sarcolemmal and cytosolic fractions. Furthermore FGF-2 induced a time-dependent elevation in cardiomyocyte membrane associated PLC gamma1 and PLC beta1 activities, assayed in immunoprecipitated fractions, and moreover, increased the membrane levels of PLC beta1 and PLC beta3. Activation of PLC beta is suggestive of FGF-2-induced cross-talk between FGF-receptor tyrosine kinase and G-protein-coupled signaling in adult cardiomyocytes and underscores the importance of FGF-2 in cardiac physiology. PMID- 10668636 TI - The expression of transforming growth factor-beta and interleukin-1beta mRNA and the response to 1,25(OH)2D3' 17 beta-estradiol, and testosterone is age dependent in primary cultures of mouse-derived osteoblasts in vitro. AB - The aim of the present study was to examine the hypothesis that primary cultures of osteoblasts obtained from bones of young animals respond to hormones better than cell cultures obtained from old animals. We studied in cultured osteoblastic cells the effects of 1,25(OH)2D3 and sex steroid hormones on several mouse osteoblastic phenotypic expressions including transforming growth factor-beta (TGF-beta) and interleukin-1beta (IL-1beta) mRNAs. Second passages of long bone derived osteoblastic cells from young donors (5-12 wk) and old donors (10-12 mo old) were used for this study. The cells obtained from old animals had decreased ALP activity and cAMP compared with cells obtained from young animals with no change in collagen production and mineralization. The addition of 17beta estradiol and testosterone increased ALP activity and mineralization in the cultured cells from both age groups and collagen production in cells obtained from old mice. Using in situ hybridization IL-1beta and TGF-beta mRNA expression was observed to be higher in the osteoblasts from young than from old donors. 1,25(OH)2D3 increased IL-1beta mRNA expression in the cells derived from young mice. Testosterone and 17beta-estradiol inhibited IL-1beta mRNA expression only in cells derived from young mice. Sex steroid hormones did not change TGF-beta mRNA expression in any of the cell lines, but 1,25(OH)2D3 increased its expression in cells derived from old donors. The results of the present study indicate that cells obtained from old mice are generally less active than those obtained from young animals. PMID- 10668635 TI - DHEA and the skeleton (through the ages). AB - Dehydroepiandrosterone (DHEA) and its sulfate ester, DHEAS, are the most abundant steroids in the human circulation, although their exact biological significance is not completely understood. DHEA(S) levels are high in fetal life, decrease after birth, and show a marked pubertal increase to a maximal level during young adulthood. In healthy adults, DHEAS levels decline to 10-20% of peak levels by age 70 yr. This review summarizes information concerning the role of DHEA in skeletal physiology, including modulation of the skeletal insulin-like growth factor regulatory system, and its effects on secretion of proresorptive cytokines. The pattern of secretion of DHEA throughout the life cycle is discussed, as well as its potential usefulness in specific disease states as an agent with anabolic and antiosteolyic effects on bone. PMID- 10668637 TI - Androgen receptor mRNA expression in the rhesus monkey ovary. AB - Immunocytochemical detection of androgen receptors (ARs) in several compartments of the macaque ovary, including the germinal epithelium, follicle, and corpus luteum, suggests a role for androgens in modulating ovarian function via the classical receptor-mediated pathway. To examine AR mRNA expression in the rhesus monkey ovary, total RNA was isolated from whole ovaries, the germinal epithelium enriched cortical and medullary compartments of the ovary, and corpora lutea from early (d 3-5), mid (d 6-8), mid-late (d 10-12), and late (d 13-15) stages of the luteal phase of the menstrual cycle. RNA was also obtained from luteinized granulosa cells from monkeys receiving gonadotropin treatment to stimulate the development of multiple ovarian follicles. After reverse transcription of total RNA using oligo-dT as a primer, polymerase chain reaction (PCR) was used to amplify a unique 329 bp segment of the monkey AR hormone-binding region. Reverse transcriptase (RT)-PCR products of the expected size were detected in all ovarian and control tissues. Sequence analysis of the AR cDNA from the macaque ovary revealed 99% nucleotide homology and 100% predicted amino acid homology to the cDNA for the hormone-binding region of human AR. Northern analysis demonstrated the presence of a major AR mRNA species at 9.5 kb in corpus luteum, luteinized granulosa cells, and prostate, with additional bands detected in the corpus luteum and prostate at 7.9 and 3.4 kb, respectively. A sensitive RNase protection assay was used to examine AR mRNA levels in ovarian tissues and showed AR mRNA expression throughout the life-span of the corpus luteum. Thus, detection of AR mRNA in the primate ovary, including the periovulatory follicle and corpus luteum, supports the concept that these tissues are targets for receptor-mediated androgen action during the menstrual cycle. PMID- 10668638 TI - Antisteroidogenic action of nitric oxide on human corpus luteum in vitro: mechanism of action. AB - To analyze the mechanism by which nitric oxide (NO) exerts its antisteroidogenic action, human luteal cells were cultured during 24 and 48 h with L-arginine (L Arg, 1 mmol/L); 1,2(2-trifluoromethylphenyl)imidazole (TRIM) (50 micromol/L and 1 mmol/L) and cyclic guanosine monophosphate (cGMP) analog (8-Br-cGMP, 1 mmol/L). Estradiol, nitrite, and P450 AROM activity were determined in culture media. Total cGMP concentration was evaluated in the cells and culture media by radioimmunoassay, and NADPH diaphorase was used as a histochemical marker for NO synthase (NOS) activity. During the corpus luteum (CL) life-span, NO affected estradiol secretion in an age-dependent manner, with an inhibition in mid-CL (37%; p < 0.05) in agreement with our previous results, and no significant modification in early and late CL. Basal nitrite concentration in 24 and 48 h of midluteal cell cultures (42 and 93 pmol/10(6) cells, respectively) was increased by L-Arg (53% and 88%) and inhibited by the two TRIM concentrations; also, an intense diaphorase reactivity was observed in endothelial cells and luteal parenchyma. Total cGMP was not detected in cell cultures and 8-Br-cGMP did not modify estradiol secretion, whereas aromatase activity was strongly inhibited by L-Arg (70%, p < .05). These results suggest that both NOS isoforms are active in midluteal cells, and the mechanism of action for NO on in vitro estradiol secretion may be an inhibition of P450 AROM activity. PMID- 10668639 TI - The effect of hypoxia on plasma leptin and insulin in newborn and juvenile rats. AB - Hypoxia leads to a decrease in food intake and attenuated weight gain in rats. The purpose of this study was to measure plasma leptin and insulin in young rats exposed to hypoxia for 7 d as compared to a normoxic control group of the same age. One group was exposed from birth to 7 d of age; the other was exposed from 28 to 35 d of age (weaned at 21 d of age). As expected, body weight was significantly lower in rats of either age exposed to hypoxia for 7 d. Plasma leptin was significantly lower in hypoxic (2.0+/-0.2 ng/mL; n = 41) compared with normoxic (2.6+/-0.3 ng/mL; n = 30) 7-d-old rats. Plasma leptin was also significantly lower in hypoxic (1.1+/-0.1 ng/mL; n = 20) as compared to normoxic (1.5+/-0.1 ng/mL; n = 20) 35-d-old rats. Seven-day-old rats exposed to hypoxia demonstrated significant increases in plasma glucose and insulin whereas 35-d-old rats exhibited a decrease in both variables. We conclude that exposure to hypoxia for 7 d leads to a decrease in body weight and plasma leptin in infant and juvenile rats. The decrease in leptin may be an attempt to reverse hypoxia induced anorexia. PMID- 10668640 TI - Antioxidative mechanisms and plasma growth hormone levels: potential relationship in the aging process. AB - Factors affecting longevity are complex and poorly understood. We have recently found that Ames dwarf mice (df/df), which are deficient in growth hormone (GH), prolactin, and thyroid-stimulating hormone, live significantly longer than their normal siblings whereas transgenic mice that overexpress GH exhibit reduced life spans and various indices of premature aging. The production of reactive oxygen species increases with aging and is associated with DNA damage to the tissues. However, several cellular oxygen scavenging/detoxifying systems exist that improve the antioxidative defense capacity of cells. We evaluated the activity of enzymes involved in this defense system in liver, kidney, and heart tissue from dwarf, phosphoenol-pyruvate carboxykinase-bovine GH transgenic, and corresponding groups of normal mice. Liver glutathione and ascorbate levels were lower (p < 0.0025) in dwarf animals compared to normal and GH transgenic mice. By contrast, the level of catalase activity, which detoxifies hydrogen peroxide, in dwarf liver and kidney was significantly higher when compared to the other groups. Animals deficient in GH (dwarf) live longer and exhibit enzyme activities and levels that may combat oxidative stress more efficiently than normal mice and those overexpressing GH. PMID- 10668641 TI - Homologous upregulation of GnRH receptor mRNA by continuous GnRH in cultured rat pituitary cells. AB - The present study examined the effects of continuous treatment with gonadotropin releasing hormone (GnRH) on GnRH receptor (GnRH-R) mRNA levels in dispersed cultures of rat pituitary cells. Pituitary GnRH-R mRNA levels were determined by competitive reverse transcriptase polymerase chain reaction. When pituitary cells were continuously exposed to a low dose of GnRH (0.2 nM), GnRH-R mRNA levels were transiently increased. The levels of GnRH-R mRNA were significantly increased up to 6 h and diminished to untreated levels by 24 h. Luteinizing hormone (LH) release was also increased significantly up to 12 h, maintaining similar levels in LH release thereafter. When GnRH antagonist ([D-pGlu1, D-Phe2, D-Trp3,6]-LH RH) was added to the cultures together with GnRH (0.2 nM) for 6 h, the stimulatory effect of GnRH on GnRH-R mRNA levels and LH release was significantly diminished in a dose-related manner. In another experiment, pituitary cells were treated with various doses of GnRH (0.02-200 nM) for a relatively short (6 h) or a longer (24 h) period. When pituitary cells were exposed for 6 h, all doses of GnRH (0.02-200 nM) significantly increased GnRH-R mRNA levels in a dose-dependent manner. By contrast, continuous exposure to GnRH for 24 h was ineffective in changing pituitary GnRH-R mRNA levels at any given doses. These results indicate that the duration of GnRH treatment is critical for upregulation of GnRH-R mRNA by continuous GnRH. When pituitary cells were treated for 6 h with either a continuous mode of GnRH (0.2 nM) or an hourly pulsatile mode of GnRH (0.2 nM, 6 min/h), both treatments significantly augmented GnRH-R mRNA levels. Thus, the modes of GnRH application, if treated for a relatively short period, do not appear to make a significant difference in upregulation of GnRH-R mRNA levels. Collectively, our data provide strong evidence that continuous GnRH application is able to upregulate pituitary GnRH-R mRNA levels, if treated for a relatively short period (6 h). PMID- 10668642 TI - Enhanced platelet aggregation, high homocysteine level, and microvascular disease in diabetic muscle infarctions: implications for therapy. AB - Muscle infarction is a rare complication in patients with diabetes mellitus, probably because of the rich vascular supply of this tissue. We describe a patient with type 1 diabetes who had infarction of the muscles in her right thigh. We report, for the first time, that the patient, in addition to an advanced microvascular disease in the muscle, had increased plasma total homocysteine levels and increased platelet aggregation. These pathologies might have a synergistic effect on the development of this rare complication and should be treated aggressively to prevent further episodes. PMID- 10668643 TI - Testicular abnormalities in male rats after lactational exposure to nonylphenols. AB - Lactational exposure of male rat pups to nonylphenols (NPs) decreased the size of their testes and male accessory glands. At 31 d of age, NP-treatment of male rats resulted in less cellular differentiation of the seminiferous tubules (STs) and increased intertubular space compared to controls. At maturity, NP-treated males showed varying degrees of abnormalities in the affected testes. In the moderately affected ones, about 20-30% of their STs had poorly differentiated germinal elements. Cell lineage was less organized. In extreme cases, all STs of the affected testis failed to differentiate into germinal elements. These abnormalities in germinal element differentiation might be the primary cause for a number of the NP-treated males having a lower epididymal sperm count and a lower percentage of motile sperm compared to age-matched control males. Zymogram analysis of testis homogenates by sodium dodecyl sulfate gelatin gels revealed two major forms (64-66 kDa and 50-52 kDa) of gelatinases. Only the 50-52-kDa form was greatly reduced or absent in the affected testis. Lactational exposure of male pups to NPs thus leads to various testicular abnormalities including lack of differentiation of STs, lowering of sperm count, and reduction in the percentage of motile sperm and modulation of a specific form of testicular proteinases. PMID- 10668644 TI - TGF-alpha exerts biphasic effects on estrogen--and phytoestrogen-mediated gene expression in breast cancer cells. AB - Transforming growth factor-alpha (TGF-alpha) contributes to the progression of mammary carcinogenesis in part through synergistic augmentation of estradiol (E2) action. To investigate this further, we sought to determine (1) whether the duration of TGF-alpha treatment might influence the nature of the TGF-alpha/E2 interaction, and (2) whether TGF-alpha would behave in a similar manner when combined with phytoestrogens. To this end, we transfected T47-D breast cancer cells with an estrogen-responsive reporter and then treated the cells (for 4-48 h) with varying concentrations of TGF-alpha, E2, the antiestrogen 4-hydroxy tamoxifen (HOT), and/or one of three phytoestrogens. Our findings revealed that TGF-alpha has short-term synergistic and long-term inhibitory effects on E2- and phytoestrogen-regulated gene expression. Furthermore, this secondary inhibition of E2 action by TGF-alpha was similar in magnitude to that imposed by HOT. These findings demonstrate a novel role for TGF-alpha and invite reevaluation of current models regarding TGF-alphas interactions with E2 in breast cancer cells. Our results also raise the possibility that phytoestrogens, which interact with TGF-alpha in a manner conceptually identical to that of E2, may subserve a regulatory function in breast cancer cells. PMID- 10668645 TI - Inhibition of hypothalamic GnRH secretion in the ewe by antigonadotropic decapeptide during the estrous cycle and nonbreeding season. AB - Previous experiments from our laboratory and others have shown that the peptide antigonadotropic decapeptide (AGD) has marked inhibitory effects on luteinizing hormone (LH) secretion in rats and ewes. The first objective of this study was to determine whether AGD inhibits LH secretion by regulating hypothalamic release of gonadotropin hormone (GnRH). AGD (200 microg in 200 microL of 0.3% bovine serum albumin [BSA] saline) or vehicle was infused into the lateral ventricle of ovariectomized (OVX) ewes with hypophyseal-portal cannulae, and GnRH secretion was monitored. The frequency of GnRH and LH pulses in AGD-treated ewes was significantly decreased (p < 0.05) but did not change in the control ewes. The second objective of this investigation was to evaluate changes in hypothalamic sensitivity to AGD in the ewe during the estrous cycle and nonbreeding season. During the estrous cycle, the effects of AGD on LH secretion were assessed following ovariectomy, during the metestrous, diestrous, and proestrous phases of the estrous cycle. The response to AGD during the estrous cycle was compared to its effect during the anestrous season. LH, cortisol, and prolactin (PRL) concentrations were assayed in peripheral blood samples obtained at 10-min intervals over a 6-h period prior to injection of either vehicle (200 microL of 0.3% BSA in 0.9% saline) or AGD (200 microg in 200 microL of vehicle), and for an additional 10 h following treatment. LH pulse frequency decreased after treatment with AGD (p < 0.05) at all times in OVX and intact ewes compared to vehicle treated controls. During the anestrous season, AGD treatment was more effective in inhibiting LH pulse frequency than during the breeding season (p < 0.05). Furthermore, there was a significant increase (p < 0.05) in mean cortisol concentrations after AGD infusion in all AGD-treated groups compared to controls independent of season or reproductive status. PRL concentrations were also increased (p < 0.05) following treatment with AGD. These results suggest that inhibition of pulsatile LH release induced by AGD is modulated by alterations in frequency of hypothalamic discharges of GnRH. Furthermore, changes in the inhibitory actions of AGD may contribute to the seasonal regulation of hypothalamic GnRH secretion in the ewe. PMID- 10668646 TI - Mutations in the gene encoding the alpha-subunit of the Gs protein in molar pregnancy. AB - Molar pregnancy is a gestational trophoblastic disease associated with a trophoblastic proliferation and a protein synthesis alteration. It is characterized by the presence of hydatiform moles, which are fluid-filled cysts derived from the chorionic villi of the placenta. Recent studies have reported a reduced expression of several types of G proteins including Gsalpha in molar pregnancies suggesting alterations in G protein structure in hydatiform moles. To identify mutations that lead to Gsalpha deficiency, we isolated genomic DNA from hydatiform moles and used polymerase chain reaction to amplify all exons of the Gsalpha gene. Amplified Gsalpha gene fragments were analyzed by sequencing using the dideoxy chain termination method. Tissues obtained from three complete hydatiform moles and one partial hydatiform mole were examined. We have identified a heterozygous 8-bp deletion in exon 10 of the Gsalpha gene, in two complete hydatiform moles, that had evidence for a dysfunctional Gsalpha protein. This deletion produced a truncated protein. We have also identified a heterozygous polymorphism in exon 5 in two complete hydatiform moles, and a homozygous substitution (A-->G) in intron 5 of the Gsalpha gene in the other complete hydatiform mole; these two last types of mutations should not have any effects on protein activity. PMID- 10668647 TI - Expression of prolactin receptor mRNA is increased in the preoptic area of lactating rats. AB - This study investigated expression of prolactin receptor (PRL-R) mRNA in the preoptic area in midlactating rats compared with diestrous rats. Tissues from specific nuclei were micropunched from 300-microm thick rat brain sections with 300- or 500-microm diameter needles. After total RNA was extracted, the two forms of PRL-R mRNA were evaluated by reverse transcriptase polymerase chain reaction and Southern hybridization. The results showed that levels of long-form PRL-R mRNA in the ventrolateral preoptic nucleus and lateroanterior nucleus in lactating rats were significantly higher (p < 0.05) than in diestrous rats. The ventromedial and medial preoptic nuclei in lactating rats also expressed moderately high levels of long-form mRNA when compared with (p = 0.0547) diestrous rats. The ventromedial and ventrolateral preoptic nuclei, and ventrolateral hypothalamic nucleus in lactating rats expressed significantly higher levels of short-form mRNA than in diestrous rats. The increased expression of both forms of PRL-R mRNA helps explain numerous effects of PRL on brain functions during lactation. PMID- 10668648 TI - Nitric oxide and hydroperoxide affect islet hormone release and Ca(2+) efflux. AB - We have investigated the influence of the intracellular free radical donors hydroxylamine (giving nitric oxide [NO]) and tert-butylhydroperoxide (giving hydroperoxide ["H2O2"]) on glucose- and cyclic adenosine monophosphate (cAMP) induced transduction signaling in islet hormone release. Both donors dose dependently inhibited glucose-stimulated insulin release and induced modest (hydroxylamine) or profound (tertbutylhydroperoxide) suppression of 45Ca2+-efflux from perifused islets. By contrast, both donors stimulated glucagon release. Similar effects on hormone release were displayed after K+-depolarization. Insulin and glucagon release stimulated by activation of the cAMP system through isobutylmethylxanthine (IBMX) at basal glucose was modestly potentiated by low concentrations of both donors. These effects were still observed, although less pronounced, in K+-depolarized islets. In vitro as well as in vivo, the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester inhibited IBMX-induced glucagon release, but did not affect insulin release. The results suggest that NO and hydroperoxide inhibit glucose-stimulated insulin release by perturbing Ca2+ fluxes and probably acting through S-nitrosylation (NO) or oxidation (hydroperoxide) of thiol groups critical to the secretory process. These effects are largely independent of depolarization events. By contrast, both NO and hydroperoxide can potentiate cAMP-stimulated hormone release presumably at a distal site in the stimulus-secretion coupling. PMID- 10668649 TI - Cross-modal generalization effects of training noncanonical sentence comprehension and production in agrammatic aphasia. AB - The cross-modal generalization effects of training complex sentence comprehension and complex sentence production were examined in 4 individuals with agrammatic Broca's aphasia who showed difficulty comprehending and producing complex, noncanonical sentences. Object-cleft and passive sentences were selected for treatment because the two are linguistically distinct, relying on wh-and NP movement, respectively (Chomsky, 1986). Two participants received comprehension training, and 2 received production training using linguistic specific treatment (LST). LST takes participants through a series of steps that emphasize the verb and verb argument structure, as well as the linguistic movement required to derive target sentences. A single-subject multiple-baseline design across behaviors was used to measure acquisition and generalization within and across sentence types, as well as cross-modal generalization (i.e., from comprehension to production and vice versa) and generalization to discourse. Results indicated that both treatment methods were effective for training comprehension and production of target sentences and that comprehension treatment resulted in generalization to spoken and written sentence production. Sentence production treatment generalized to written sentence production only; generalization to comprehension did not occur. Across sentence types generalization also did not occur, as predicted, and the effects of treatment on discourse were inconsistent across participants. These data are discussed with regard to models of normal sentence comprehension and production. PMID- 10668650 TI - The effects of inflectional variation on fast mapping of verbs in English and Spanish. AB - To use morphological cues for syntactic bootstrapping, children must recognize that inflectionally varying words (e.g., pushes, pushed) are instances of the same word. Children who are exposed to languages with richer inflectional morphologies than English, such as Spanish, encounter instances of inflectional variation more often. Thus they may learn to recognize inflectionally varying words as instances of the same word at an earlier age than do learners of English. English-and Spanish-learning 3-year olds were taught novel verbs in a fast mapping task under two conditions: no-inflectional variation in which inflections did not vary between exposure and testing (e.g., neps, neps) and inflectional variation in which inflections alternated between exposure and testing (e.g., neps, nepped). Children's scores were significantly higher in the no-variation condition than in the variation condition. There were no significant differences between the performance of the language groups. These findings suggest that even children acquiring languages with relatively rich verbal inflection paradigms may not be able to consistently parse stems and inflections to associate inflectionally varying forms. PMID- 10668651 TI - Narrative production by children with and without specific language impairment: oral narratives and emergent readings. AB - The research reported in this paper was based on the premise that oral and written language development are intertwined. Further, the research was motivated by research demonstrating that narrative ability is an important predictor of school success for older children with language impairment. The authors extended the inquiry to preschool children by analyzing oral narratives and "emergent storybook reading" (retelling of a familiar storybook) by two groups of 20 children (half with, half without language impairment) age 2;4 (years;months) to 4;2. Comparative analyses of the two narrative genres using a variety of language and storybook structure parameters revealed that both groups of children used more characteristics of written language in the emergent storybook readings than in the oral narratives, demonstrating that they were sensitive to genre difference. The children with language impairment were less able than children developing typically to produce language features associated with written language. For both groups, middles and ends of stories were marked significantly more often within the oral narratives than the emergent readings. The children with language impairment also had difficulty with other linguistic features: less frequent use of past-tense verbs in both contexts and the use of personal pronouns in the oral narratives. Emergent storybook reading may be a useful addition to language sampling protocols because it can reveal higher order language skills and contribute to understanding the relationship between language impairment and later reading disability. PMID- 10668652 TI - The role of phonological opacity in reading achievement. AB - This study investigated the relation among phonological awareness, morphological awareness, and reading achievement in 69 children with and without language learning disabilities. Children participated in two morphological tasks that assessed skill in identifying the bases and suffixes of phonologically transparent and opaque derivatives. Transparent derivatives preserve the phonological characteristics of the base word (e.g., allow-allowable, pure purist); opaque derivatives involve stress and/or vowel changes to the base (e.g., acid-acidic, flame-flammable). Children with language-learning disabilities were outperformed by chronological-age peers on each task and showed a level of accuracy similar to that of younger, typically achieving children. Regression analyses were used to determine the proportion of variance in reading accounted for by the morphological tasks beyond that accounted for by age and vocabulary knowledge. Performance with transparent derivatives added a significant, but small, proportion (6.9%) to total variance in word identification scores and a nonsignificant proportion (2.2%) to passage comprehension scores. Performance with opaque derivatives added a substantial contribution to word-identification scores (19.9%) and passage-comprehension scores (16.5%) beyond that accounted for by age, vocabulary knowledge, and performance with transparent derivatives. These results suggest that the ability to analyze phonological changes associated with derivation may mediate much of the link between the type of morphological awareness assessed here and reading achievement. PMID- 10668653 TI - Mechanisms of discourse comprehension impairment after right hemisphere brain damage: suppression in lexical ambiguity resolution. AB - Normal comprehension skill is linked with the proficiency of a suppression mechanism, which functions to dampen mental activation that becomes irrelevant or inappropriate to a final interpretation. This study investigated suppression and discourse comprehension in adults with right brain damage (RBD). To index suppression function, 40 adults with RBD and 40 without brain damage listened to sentence stimuli that biased the meaning of a sentence-final lexical ambiguity (e.g., SPADE), then judged whether a probe word (e.g., CARDS) fit the overall sentence meaning. Probes represented the contextually inappropriate meanings of the ambiguities and were presented in two conditions: 175 ms and 1000 ms post stimulus. The same probes were used with unambiguous comparison stimuli. Probe judgment response times indicated that only the group without brain damage suppressed inappropriate interpretations over time. In a multiple regression analysis, suppression function added significantly to predicting performance on a general measure of narrative discourse comprehension for participants with RBD. The discussion addresses how suppression deficits may account more broadly for comprehension difficulties after RBD; it also considers several unresolved issues concerning the suppression construct and the suppression deficit hypothesis. PMID- 10668654 TI - Risk for speech disorder associated with early recurrent otitis media with effusion: two retrospective studies. AB - The goals of this two-part series on children with histories of early recurrent otitis media with effusion (OME) were to assess the risk for speech disorder with and without hearing loss and to develop a preliminary descriptive-explanatory model for the findings. Recently available speech analysis programs, lifespan reference data, and statistical techniques were implemented with three cohorts of children with OME and their controls originally assessed in the 1980s: 35 typically developing 3-year-old children followed since infancy in a university affiliated pediatrics clinic, 50 typically developing children of Native American background followed since infancy in a tribal health clinic, and (in the second paper) 70 children followed prospectively from 2 months of age to 3 years of age and older. Dependent variables included information from a suite of 10 metrics of speech production (Shriberg, Austin, Lewis, McSweeny, & Wilson, 1997a, 1 997b). Constraints on available sociodemographic and hearing status information limit generalizations from the comparative findings for each database, particularly data from the two retrospective studies. The present paper reports findings from risk analysis of conversational speech data from the first two cohorts, each of which included retrospective study of children for whom data on hearing loss were not available. Early recurrent OME was not associated with increased risk for speech disorder in the pediatrics sample but was associated with approximately 4.6 (CI = 1.10-20.20) increased risk for subclinical or clinical speech disorder in the children of Native American background. Discussion underscores the appropriateness of multifactorial risk models for this subtype of child speech disorder. PMID- 10668656 TI - Observed and reported expressive vocabulary and word combinations in bilingual toddlers. AB - The consistency of parental reports of expressive vocabulary and word combinations with observed expressive language among 21- to 27-month-old children exposed to English and Spanish on a regular basis was the focus of this investigation. Parental reports were obtained using the Spanish-English Vocabulary Checklist (Patterson, 1998), an adaptation of the Language Development Survey (Rescorla, 1989). The number of different words parents reported was correlated (r = .66) with the number of different words the children used during a 30-min videotaped interaction with the reporting parent. Parental reports of whether the child was combining words and estimates of proportion of the child's use of Spanish and English also were consistent with the children's language use during the 30-min language samples. PMID- 10668655 TI - Otitis media, fluctuant hearing loss, and speech-language outcomes: a preliminary structural equation model. AB - The goals of this study were to estimate the risk for lowered speech-language outcomes associated with early recurrent otitis media with effusion (OME) with and without hearing loss and to develop a preliminary descriptive-explanatory model for the findings. Three statistical approaches were used to assess associations among OME, hearing loss, and speech-language outcomes. Participants were a subsample of 70 children followed prospectively in the Dallas Cooperative Project on Early Hearing and Language Development (Friel-Patti & Finitzo, 1990). Findings indicated that hearing levels at 12-18 months were significantly associated with speech delay and low language outcomes at 3 years of age. The risk for subclinical or clinical speech delay at 3 years of age was 2% for children with less than 20 dB average hearing levels at 12-18 months and 33% for children with greater than 20 dB average hearing levels at 12-18 months. A structural equation model (Joreskog & Sorbom, 1993) indicated that the significant and substantial effects of hearing levels at 12-18 months on speech status at 3 years were significantly mediated by language status at 3 years. Discussion includes implications of these findings for alternative speech perception models linking early OME and hearing loss to later speech-language disorder. PMID- 10668657 TI - Adults' perception and production of the English vowel /i/. AB - This study investigated the link between the perception and production of the English vowel /i/ by adult native speakers of English. Participants first produced the vowel /i/ using normal (citation) and careful (hyperarticulated) speech, then completed a method of adjustment task in which they selected their ideal exemplar of /i/. In this perceptual task, 24 of 35 participants had a prototype; the remaining 11 did not, but were retained for comparison. In keeping with the hyperspace effect (K. Johnson, E. Flemming, & R. Wright, 1993), all participants selected perceptual stimuli with F1 and F2 values that were more extreme (i.e., higher and further forward in the vowel space) than those of their normal, citation productions. An analysis of front-back and high-low qualities for the perceptual and production data in Euclidian space revealed that hyperarticulated speech was closer to the perceptual data than citation speech was, but only for participants with relatively clear-cut prototypes. The basis for such individual variation in perception-production links is discussed. PMID- 10668658 TI - Children's phoneme identification in reverberation and noise. AB - This study assessed the effects of reverberation, noise, and their combination on listeners' identification of consonants and vowels in naturally produced nonsense syllables presented at different sensation levels (re: speech recognition threshold). A secondary purpose of this study was to assess listeners' identification of voicing, manner, and place of articulation for consonants at 50 dB SL in the reverberation, noise, and combined conditions. Listeners, aged 6-30 years, identified consonant-vowel-consonant-vowel (CVCV) stimuli presented at four different sensation levels (re: speech recognition threshold) of 30, 40, 50, and 60 dB SL in 4 listening conditions: (a) an optimal listening situation (no reverberation, no noise), (b) reverberation only (1.3 seconds), (c) noise only (+13 dB S/N against a multitalker babble), and (d) reverberation plus noise. Results showed that all listener groups achieved maximum consonant identification performance at 50 dB SL. Vowel identification scores were unaffected by SL. Statistical analyses revealed that children's ability to identify consonants varied according to listening condition. For example, children's consonant identification abilities reached adult-like levels of performance at about age 14 years in the reverberation-only and noise-only listening conditions. However, in the reverberation-plus-noise listening condition, children's consonant identification abilities do not mature until the late teenage years. The ability to identify vowels, on the other hand, develops much earlier. A feature analysis of the consonant data showed that for all 3 features (voicing, manner, and place), identification scores were highest in the control condition, similar for the reverberation-only and noise-only conditions, and lowest in the reverberation plus-noise condition. Voicing was easier for listeners to identify than manner or place of articulation features in reverberation and noise. Taken together, these results suggest that the ability to identify speech in reverberation and noise reaches adult-like level of performance at different ages for different components of the speech signal. PMID- 10668659 TI - Effects of facial paralysis and audiovisual information on stop place identification. AB - This study investigated how listeners' perceptions of bilabial and lingua alveolar voiced stops in auditory (A) and audiovisual (AV) presentation modes were influenced by articulatory function in a girl with bilateral facial paralysis (BFP) and a girl with normal facial movement (NFM). The Fuzzy Logic Model of Perception (FLMP) was used to make predictions about listeners' identifications of stop place based on assumptions about the nature (clear, ambiguous, or conflicting) of the A or AV cues produced by each child during /b/ and /d/ CV syllables. As predicted, (a) listeners' identification scores for NFM were very high and reliable, regardless of presentation mode or stop place, (b) listeners' identification scores for BFP were high for lingua-alveolar place, regardless of presentation mode, but more variable and less reliable than for NFM; significantly lower (overall at a chance level) for bilabial place in the A mode; and lowest for bilabial place in the AV mode. Conflicting visual cues for stop place for BFP's productions of /bV/ syllables influenced listeners' perceptions, resulting in most of her bilabial syllables being misidentified in the AV mode. F2 locus equations for each child's /bV/ and /dV/ syllables showed patterns similar to those reported by previous investigators, but with less differentiation between stop place for BFP than NFM. These acoustic results corresponded to the perceptual results obtained. (That is, when presented with only auditory information, on average, listeners perceived BFP's target /b/ syllables to be near the boundary between /b/ and /d/.) PMID- 10668660 TI - The use of tactile supplements in lipreading Swedish and English: a single subject study. AB - The speech perception skills of GS, a Swedish adult deaf man who has used a "natural" tactile supplement to lipreading for over 45 years, were tested in two languages: Swedish and English. Two different tactile supplements to lipreading were investigated. In the first,"Tactiling," GS detected the vibrations accompanying speech by placing his thumb directly on the speaker's throat. In the second, a simple tactile aid consisting of a throat microphone, amplifier, and a hand-held bone vibrator was used. Both supplements led to improved lipreading of materials ranging in complexity from consonants in [aCa] nonsense syllables to Speech Tracking. Analysis of GS's results indicated that the tactile signal assisted him in identifying vowel duration, consonant voicing, and some manner of articulation categories. GS's tracking rate in Swedish was around 40 words per minute when the materials were presented via lipreading alone. When the lipreading signal was supplemented by tactile cues, his tracking rates were in the range of 60-65 words per minute. Although GS's tracking rates for English materials were around half those achieved in Swedish, his performance showed a similar pattern in that the use of tactile cues led to improvements of around 40% over lipreading alone. PMID- 10668661 TI - Validation assessment of a French version of the tinnitus reaction questionnaire: a comparison between data from English and French versions. AB - The present study compares the results obtained on original and French versions of the TRQ (Tinnitus Reaction Questionnaire) initially published by Wilson, Henry, Bowen, and Haralambous (1991) in English to evaluate the psychological distress of tinnitus sufferers. Reliability and validity of the French translation were determined using data from 173 normal hearing or hearing impaired patients with tinnitus lasting from 1 month to 41 years. They completed the translated questionnaire and a short version of the Minnesota Multiphasic Personality Inventory. The results indicated good internal consistency (Cronbach's alpha = .94), and the reliability of the French version of the TRQ was demonstrated, except for items 5 and 20. High statistically significant correlations were found between the TRQ and Depression, Psychaesthenia, and Anxiety Mini-Mult subscales. The validation demonstrates only minor effects of language. The French version of the TRQ thus is an equally valid tool as the original English version for evaluating tinnitus distress of a patient. PMID- 10668662 TI - Acoustic and airflow spectral analysis of voice tremor. AB - Acoustic spectral analysis has been used to describe voice tremor with some success, but no feature distinguishing pathological from normal tremor has been clearly identified. To assist in monitoring voice tremor associated with neurological diseases, objective and quantifiable measures that can distinguish between normal and pathological tremor are desired. This study explored the plausibility of using airflow and acoustic signals to quantify the frequency and amplitude of voice tremor and potentially to distinguish pathological from normal tremor. Subjects were 10 individuals with pathological tremor, most of them individuals with Parkinson's disease, and 10 gender and age-matched individuals with no voice disorder. Simultaneous acoustic and airflow signals were recorded during sustained vowel phonation. The acoustic intensity contours and the airflow signals were submitted to spectral analysis. A peak prominence ratio, defined as the ratio of the spectral peak energy to the overall signal energy, was calculated for each spectral peak below 30 Hz. For each subject, the 6 spectral peaks with the highest peak prominence ratios were selected. Frequency values of the 6 selected acoustic or airflow spectral peaks failed to distinguish tremor group from control group. Peak prominence ratios of the 6 selected acoustic spectral peaks were significantly higher for tremor group than for control group. Although spectral analysis of airflow signals was not useful in differentiating tremor group from control group, acoustic intensity contours and airflow time waveforms were highly and positively correlated in more tremor subjects (90%) than control subjects (40%). This finding suggests that the relationship between acoustic intensity contours and airflow time waveforms may reflect the presence and the source of voice tremor. PMID- 10668663 TI - Patterns of orofacial movement velocity across variations in speech rate. AB - To understand the clinical aspects of speech rate control, a clearer picture is needed of how orofacial structures are coordinated across variations in speech rate. To address this problem, patterns of orofacial tangential velocity or speed were analyzed in a group of 9 normal speakers as they produced the utterance "a bad daba" at fast, normal, and slow speech rates. An electromagnetic system was used to record the movements of the upper lip, lower lip, jaw, and tongue. Measures of the magnitude of peak tangential velocities were obtained across the four structures. Orofacial velocities consistently decreased at slow rates relative to normal rates, whereas at fast rates increased and decreased velocities were observed in an equivalent number of cases. Significant correlations frequently were obtained across speech rate between lip, tongue, and jaw velocities. Upper and lower lip velocities showed consistent positive correlations with one another, whereas marked intersubject differences were observed in the sign of jaw-related correlations. Repeated testing on 3 subjects indicated a high degree of consistency within subjects in the overall patterns of mean velocity for the different structures. Results are discussed in relation to possible motor control differences underlying fast and slow speech, neural coupling of muscle systems, and jaw-related individual differences in speech motor coordination. PMID- 10668664 TI - Frequency and temporal resolution in elderly listeners with good and poor word recognition. AB - There is a subgroup of elderly listeners with hearing loss who can be characterized by exceptionally poor speech understanding. This study examined the hypothesis that the poor speech-understanding performance of some elderly listeners is associated with disproportionate deficits in temporal resolution and frequency resolution, especially for complex signals. Temporal resolution, as measured by gap detection, and frequency resolution, as measured by the critical ratio, were examined in older listeners with normal hearing, older listeners with hearing loss and good speech-recognition performance, and older listeners with hearing loss and poor speech-recognition performance. Listener performance was evaluated for simple and complex stimuli and for tasks of added complexity. In addition, syllable recognition was assessed in quiet and noise. The principal findings were that older listeners with hearing loss and poor word-recognition performance did not perform differently from older listeners with hearing loss and good word recognition on the temporal resolution measures nor on the spectral resolution measures for relatively simple stimuli. However, frequency resolution was compromised for listeners with poor word-recognition abilities when targets were presented in the context of complex signals. Group differences observed for syllable recognition in quiet were eliminated in the noise condition. Taken together, the findings support the hypothesis that unusual deficits in word recognition performance among elderly listeners were associated with poor spectral resolution for complex signals. PMID- 10668665 TI - Supraglottic activity: evidence of vocal hyperfunction or laryngeal articulation? AB - False vocal fold (FVF) adduction and compression of the arytenoid cartilages to the petiole of the epiglottis in an anterior to posterior (A-P) direction have been thought to characterize voice disorders with abnormally increased muscle tension or effort, often termed hyperfunctional voice disorders. To further evaluate the association between hyperfunctional voice disorders and supraglottic activity, we compared the incidence of static and dynamic supraglottic activity in individuals with normal laryngeal mucosa, normal voice quality, and no voice complaints to two populations: subjects with vocal fold nodules and subjects with complaints of dysphonia without visible vocal fold lesions, glottal incompetence, or impairment of arytenoid cartilage motion ("hyperfunctional" group). Thirty-two subjects were assigned to one of these three groups (10 control, 12 nodule, and 10 hyperfunctional). Laryngeal movements were recorded using flexible videoendoscopy while a subject was performing speech tasks such as sustained phonation, syllable repetitions, sentence imitations, and conversation. Samples were randomized by subject and task and rated for presence or absence of A-P and FVF compression. Statistically significant group differences were found for FVF compression across speech tasks (chi-square, p<0.001). The control group had the smallest incidence (45%), nodule patients the next larger incidence (68%), and hyperfunctional patients the largest incidence (80%). Statistically significant group differences were found for A-P compression across speech tasks (chi-square, p<.05). The control group had the smallest incidence (74%), nodule patients the next larger incidence (78%), and hyperfunctional patients the largest incidence (92%). Statistically significant task differences were found for the presence of FVF compression in control subjects (chi-square, p<.005), hyperfunctional patients (chi-square, p<.025), and nodule patients (chi-square, p<.001), but not for A-P compression for any of the groups. A higher incidence of FVF compression was present for the speech tasks that included glottal stops. This context specific variation in supraglottic activity suggested a dynamic component to FVF compression and also explained the high proportion of FVF compression in the control group. Each video sample was also rated for consistency of FVF or A-P compression to explore the static and dynamic nature of supraglottic activity. For samples on which raters agreed, A-P compression was typically present consistently, suggesting a static component, and FVF compression inconsistently, suggesting a dynamic component, for all three groups (chi-square, p<.001). These findings do not support previous suggestions that supraglottic activity may be a precursor to developing vocal fold nodules, as the nodule patients did not exhibit a higher incidence or consistency of A-P or FVF compression than patients with hyperfunctional voicing patterns in this study. Subjects in the hyperfunctional voice group were found to have static components of FVF and A-P compression. The presence of FVF compression in speech tasks that included glottal stops in the control group suggests an articulatory function at the laryngeal level. PMID- 10668667 TI - Strength, endurance, and stability of the tongue and hand in Parkinson disease. AB - Weakness and fatigue in the orofacial system often are presumed to contribute to the dysarthria associated with neuromotor disorders, although previous research findings are equivocal. In this study, tongue strength, endurance, and stability during a sustained submaximal effort were assessed in 16 persons with mild to severe Parkinson disease (PD) and a perceptible speech disorder. The same measures were taken from one hand for comparison. Only tongue endurance was found to be significantly lower in these participants than in neurologically normal control participants matched for sex, age, weight, and height. Analyses of data from a larger sample comprising the present and retrospective data revealed lower than-normal tongue strength and endurance in participants with PD. No significant correlations were found between tongue strength and endurance, interpause speech rate, articulatory precision, and overall speech defectiveness for the present and previously studied participants with PD, bringing into question the influence of modest degrees of tongue weakness and fatigue on perceptible speech deficits. PMID- 10668666 TI - The physiologic development of speech motor control: lip and jaw coordination. AB - This investigation was designed to describe the development of lip and jaw coordination during speech and to evaluate the potential influence of speech motor development on phonologic development. Productions of syllables containing bilabial consonants were observed from speakers in four age groups (i.e., 1-year olds, 2-year-olds, 6-year-olds, and young adults). A video-based movement tracking system was used to transduce movement of the upper lip, lower lip, and jaw. The coordinative organization of these articulatory gestures was shown to change dramatically during the first several years of life and to continue to undergo refinement past age 6. The present results are consistent with three primary phases in the development of lip and jaw coordination for speech: integration, differentiation, and refinement. Each of these developmental processes entails the existence of distinct coordinative constraints on early articulatory movement. It is suggested that these constraints will have predictable consequences for the sequence of phonologic development. PMID- 10668668 TI - Short-latency changes in voice F0 and neck surface EMG induced by mechanical perturbations of the larynx during sustained vowel phonation. AB - Nineteen healthy young adult males with normal voice and speech attempted to sustain the vowel /u/ at a constant pitch (target: 180 Hz) and a constant and comfortable loudness level while receiving a sudden mechanical perturbation to the larynx (thyroid prominence) via a servo-controlled probe. The probe moved toward or away from the larynx in a ramp-and-hold fashion (3.3-mm displacement, 0.7 N force, 20-ms rise time, 250-ms duration) as the subjects attempted to maintain a constant probe-larynx pressure. Eighty stimuli were applied in each direction, one stimulus per phonation. Pairs of surface electromyography (EMG) electrodes were attached to the skin of the anterior neck over laryngeal, infralaryngeal, and supralaryngeal areas. The rectified EMG signals, the voltage analog of the voice fundamental frequency (VAF0), and the voltage analog of the probe displacement were digitized and signal-averaged relative to the onset of the stimulus. Sudden perturbation of the larynx induced an instantaneous decrease or increase in VAF0, depending on the direction of the probe's movement, and a short-latency increase in the EMG (30-35 ms) and VAF0 (55-65 ms). We argue that the instantaneous VAF0 change was related to a mechanical effect, and the short latency VAF0 and EMG changes to reflexogenic effects-the latter most likely associated with both intrinsic and extrinsic laryngeal sensorimotor mechanisms. Further physiological studies are needed to elucidate the sources of the VAF0 and EMG responses. Once elucidated, the present method may provide a powerful noninvasive tool for studying laryngeal neurophysiology. The theoretical and clinical implications of the present findings are addressed. PMID- 10668669 TI - On the assessment of stability and patterning of speech movements. AB - Speech requires the control of complex movements of orofacial structures to produce dynamic variations in the vocal tract transfer function. The nature of the underlying motor control processes has traditionally been investigated by employing measures of articulatory movements, including movement amplitude, velocity, and duration, at selected points in time. An alternative approach, first used in the study of limb motion, is to examine the entire movement trajectory over time. A new approach to speech movement trajectory analysis was introduced in earlier work from this laboratory. In this method, trajectories from multiple movement sequences are time- and amplitude-normalized, and the STI (spatiotemporal index) is computed to capture the degree of convergence of a set of trajectories onto a single, underlying movement template. This research note describes the rationale for this analysis and provides a detailed description of the signal processing involved. Alternative interpolation procedures for time normalization of kinematic data are also considered. PMID- 10668670 TI - Otologic and audiologic evaluation of human immunodeficiency virus-infected patients. AB - PURPOSE: To quantify the incidence of ear disease in patients infected with human immunodeficiency virus (HIV). MATERIALS AND METHODS: This is a descriptive case series of HIV-positive patients, with data collected using an otologic questionnaire. otologic examination, audiologic evaluation, and chart review. The study was performed at an urban University Hospital's outpatient Infectious Disease and Otolaryngology clinics. A consecutive sample of 50 HIV-infected patients volunteered for this study. Ten subjects refused. Almost all patients received public assistance for medical care. Descriptive results were tabulated. Audiometric data were analyzed for ear, Centers for Disease Control (CDC) group, otologic complaint, and age effects. Data were compared with established norms. RESULTS: Twenty-three men and 27 women with a mean age of 40 years and mean duration of HIV disease of 3.5 years were studied. Eighteen percent of patients were in category CDC-A, 38% in CDC-B, and 44% in CDC-C. Otologic complaints were more prevalent than expected: 34% of patients reported aural fullness, 32% dizziness, 29% hearing loss, 26% tinnitus, 23% otalgia, and 5% otorrhea. Results of the neuro-otologic examination were abnormal in 33%. Tympanometric examination was abnormal in 21%. A significant degree of high-frequency sensorineural hearing loss was observed. CDC-B and CDC-C patients had worse hearing than CDC-A patients at 3 frequencies. Patients who complained of hearing loss had significantly worse otoacoustic emission results and hearing results than patients who did not, at all frequencies except 1,000 Hz. Patients in their 30s had better hearing in the speech frequencies than did all other patients. CONCLUSIONS: Ear disease affects up to 33% of HIV-infected patients. Otitis media is a frequent finding. Sensorineural hearing loss is more severe in patients with more severe HIV infection. Patients with ear complaints have demonstrable otopathology. Continuation of this preliminary descriptive work is necessary. PMID- 10668671 TI - Head and neck manifestations of non-Hodgkin's lymphoma in human immunodeficiency virus-infected patients. AB - PURPOSE: Non-Hodgkin's lymphoma is the 2nd most common malignancy in human immunodeficiency virus (HIV)-infected patients. However, limited information regarding head and neck manifestations of non-Hodgkin's lymphoma is present in the literature. The aim of this article is to describe the head and neck manifestations of non-Hodgkin's lymphoma in HIV-infected patients and compare it with that seen in noninfected patients. PATIENTS AND METHODS: A case-control study was performed including 124 patients with non-Hodgkin's lymphoma presenting over a 5.5-year period to tertiary care center in a metropolitan location. RESULTS: Overall, the anatomic distribution of non-Hodgkin's lymphoma is not altered in the presence of HIV infection with the head and neck region (63%) most often involved overall. However, within the head and neck region, extralymphatic disease is significantly more common in HIV-infected patients (59%) than noninfected patients (33%; P = .001). Central nervous system (CNS) involvement accounts for 41% of head and neck non-Hodgkin's lymphoma in HIV-infected patients, in contrast to only 12% of noninfected patients. High-grade lymphoma (68%) are more common than intermediate (30%) or low-grade disease (2%) in the HIV-infected population, whereas low (24%) and intermediate (60%) grades are more common than high-grade lymphoma (16%) in noninfected patients (P < .001). The large cell immunoblastic type (48%) is the most common subtype in HIV-infected patients, whereas diffuse large-cell type (32%) was most common in HIV-negative patients (P < .05). Survival is significantly poor for HIV-infected patients (P < .05). The impact of HIV infection on survival remain significant even after controlling for the effects of confounding factors. CONCLUSIONS: Head and neck involvement with non-Hodgkin's lymphoma occurs in a significant number of HIV infected patients. Our data show that the distribution and course of non Hodgkin's lymphoma is unique in HIV-infected patients. A high level of suspicion for non-Hodgkin's lymphoma is required in all cases of head and neck lesions in patients with HIV infection to facilitate management. PMID- 10668672 TI - Laryngeal long-term morbidity after supraglottic laryngectomy and postoperative radiation therapy. AB - PURPOSE: This study was performed to investigate factors associated with laryngeal morbidity when postoperative radiation therapy (RT) is added to supraglottic laryngectomy. MATERIALS AND METHODS: From 1980 to 1994, 56 patients affected with T1 to 4 N0 to 2c supraglottic squamous cell carcinoma selected for standard (59%) or extended (41%) supraglottic laryngectomy at 2 different institutions were retrospectively analyzed. Most of the patients (91%) also underwent neck dissection. Approximately 80% of the patients had stage T4 primary lesions or N2 neck disease. Postoperative RT was added for presumed microscopic disease at the primary site (13 patients), regional nodes (23 patients), or both (20 patients). Median delivered doses to the larynx and to the neck were 50 Gy (range, 40 to 64 Gy) and 46 Gy (range, 40 to 64 Gy), respectively. Median follow up for living patients is 11 years (range, 2.8 to 16.9 years). Laryngeal complication was defined as the appearance of grade 2 or higher toxicity according to the European Organization for Research and Treatment of Cancer (EORTC) and the Radiation Therapy Oncology Group (RTOG) scoring systems. RESULTS: Two- and 5-year actuarial locoregional control rates were 85+/-5% and 83+/-5%, respectively. Thirty patients (54%) developed laryngeal complications. However, just one patient experienced grade 4 laryngeal oedema requiring permanent tracheostomy. Estimated actuarial survival without laryngeal complications were 50+/-7%, 43+/-7%, and 39+/-7% at 2, 5, and 10 years, respectively. At univariate analysis, treated volumes (P = .03) and total dose to the larynx (P = .03) were significantly associated with local toxicity. A trend was observed also for the maximum dose to the neck (P = .06) and dose per fraction (P = .09). A multivariate Cox proportional hazards model showed total dose to the larynx to be the only independent predictor of toxicity (P = .03). The hazard ratio of laryngeal toxicity was 2.2 (95% confidence interval: 1.1/4.6), for a total dose to the larynx greater than 50 Gy. CONCLUSION: After supraglottic laryngectomy, postoperative RT to the neck does not affect local morbidity, but careful RT treatment planning is necessary to avoid delivering a total dose to the larynx greater than 50 Gy. PMID- 10668673 TI - Management of congenital laryngeal malformations. AB - Congenital malformations of the larynx are relatively rare but may be life threatening. The most common causes include laryngomalacia, vocal cord paralysis, and subglottic stenosis. The last 20 years has seen major advances in the field of surgical correction of such anomalies also serving to reduce the number of tracheotomies in children and the inherent dangers they pose. Success rates for the most popular surgical procedures have been favorable. These include supraglottoplasty for cases of severe laryngomalacia, in which relief of respiratory symptoms has been shown to occur in excess of 80% of cases. Complication rate is low, although postoperative death has been reported. Failure usually occurs in patients with concomitant airway abnormalities including pharyngomalacia. Vocal cord lateralization for vocal cord paralysis with airway compromise is achieved by means of arytenoidopexy or arytenoidectomy, using the lateral approach. Arytenoidectomy also can be performed using laryngofissure or endoscopic laser excision. Subglottic stenosis is the 3rd most common congenital anomaly. Anterior or multiple cricoid splitting with cartilage graft interpositioning is usually performed. The success rates for these procedures has been shown to be approximately 90%. PMID- 10668674 TI - Rationale for elective neck dissection. PMID- 10668675 TI - Coil embolization of a ruptured carotid pseudoaneurysm presenting as epistaxis- pediatric otolaryngology: principles and practice. PMID- 10668676 TI - Nonadjacent mass compression dizziness and hearing loss: a case report. PMID- 10668677 TI - Congenital malformations of middle and inner ears of parabiotic twins. AB - We describe herein the congenital malformations of the middle and inner ears in temporal bones of parabiotic, monozygotic twins. Temporal bones were removed from twin B, who had no fetal cardiac activity and was born dead at 23-4/7 weeks, and twin A, the donor or "pump" twin in intrauterine life, who died shortly after birth at 20-6/7 weeks. The temporal bones were processed routinely in celoidin, stained with hematoxylin and eosin, and examined by light microscopy. We found that twin B had Mondini's dysplasia with associated deformities of the middle ear and in general showed more developmental anomalies than twin A, and we conclude that Mondini's dysplasia with anomalies of the middle ear may occur in the parabiotic twin syndrome, and the abnormalities may be explainable as the result of vascular disturbance, which also causes other lesions in these unusual cases. PMID- 10668678 TI - Management of aberrant internal carotid artery injuries in children. PMID- 10668679 TI - Dermoid cysts of floor of the mouth: report of four cases. PMID- 10668680 TI - Ancient cervical vagal neurilemmoma: a case report. PMID- 10668681 TI - Chorda tympani neuroma: diagnosis and management. PMID- 10668682 TI - Fair winds and foul headaches: risk factors and triggers of migraine. PMID- 10668683 TI - The ketogenic diet: a therapy in search of an explanation. PMID- 10668684 TI - Of mice and men: a model of HIV encephalitis. PMID- 10668685 TI - Stroke in patients with heart failure and reduced left ventricular ejection fraction. AB - BACKGROUND: Cardiac failure is associated with both stroke of presumed cardioembolic origin and a high mortality rate. Warfarin is used frequently in patients with reduced cardiac left ventricular ejection fraction (EF), although no randomized trials have confirmed that anticoagulation benefits these patients. METHODS: A literature review was performed pertaining to the frequency of stroke and mortality, and the effect of antithrombotic agents on stroke and mortality rates, in patients with cardiac failure or reduced cardiac EF. We also reviewed the main features of two new proposed studies (Warfarin and Antiplatelet Therapy in Chronic Heart Failure [WATCH] and Warfarin Versus Aspirin in Reduced Cardiac Ejection Fraction [WARCEF]) comparing warfarin and antiplatelet agents in patients with low EF. RESULTS: The risk of stroke increases with decreasing EF and the risk of mortality increases with the clinical severity of cardiac failure (New York Heart Association class). Data from heart failure treatment studies suggest that warfarin may reduce stroke and mortality in patients with reduced EF, but definitive answers are lacking. The stroke rate alone is too low to be used as a primary endpoint, but an endpoint combining stroke and death (as WARCEF and WATCH propose) should allow an assessment of the effect of antithrombotics in cardiac failure. Amalgamating the data on stroke from these two trials should yield enough statistical power to compare the effects of warfarin and aspirin on stroke as an independent secondary endpoint. CONCLUSION: Whether warfarin is superior to aspirin in reducing stroke and mortality in patients with low ejection fraction is an important clinical issue that warrants prospective evaluation. PMID- 10668686 TI - Estrogen for Alzheimer's disease in women: randomized, double-blind, placebo controlled trial. AB - BACKGROUND: AD, the most prevalent cause of dementia, affects twice as many women as men. Therapeutic options are limited, but results of prior studies support the hypothesis that estrogen treatment may improve symptoms of women with this disorder. METHODS: Forty-two women with mild-to-moderate dementia due to AD were enrolled into a randomized, double-blind, placebo-controlled, parallel-group trial of unopposed conjugated equine estrogens (1.25 mg/day) for 16 weeks. RESULTS: Outcome data were available for 40 women at 4 weeks and 36 women at 16 weeks. At both 4 and 16 weeks, there were no significant differences or statistical trends between treatment groups on the primary outcome measure (the cognitive subscale of the Alzheimer's Disease Assessment Scale), clinician-rated global impression of change, or caregiver-rated functional status. Exploratory analyses of mood and specific aspects of cognitive performance also failed to demonstrate substantial group differences. CONCLUSION: Although conclusions are limited by small sample size and the possibility of a type II error, results suggest that short-term estrogen therapy does not improve symptoms of most women with AD. These findings do not address possible long-term effects of estrogen in AD, possible interactions between estrogen and other treatment modalities, or putative effects of estrogen in preventing or delaying onset of this disorder. PMID- 10668687 TI - Chinook winds and migraine headache. AB - OBJECTIVE: To determine the effects of chinook weather conditions on probability of migraine headache onset. BACKGROUND: Many migraineurs believe weather to be a trigger factor for their headaches; however, there is little supportive evidence in the literature. Migraineurs in the southern part of the Canadian province of Alberta frequently report that chinooks, warm westerly winds specific to the region, trigger their headaches. METHOD: Weather data from Environment Canada were used to designate each calendar day during the study period as a chinook, prechinook, or nonchinook day. Headache data were collected from 75 patient diaries from the University of Calgary Headache Research Clinic. Individual and multiple logistic regression models were used to determine if the weather conditions affected the probability of migraine onset. RESULTS: The probability of migraine onset was increased on both prechinook days (odds ratio 1.24; 95% CI 1.08 to 1.42) and on days with chinook winds (1.19; 1.02 to 1.39) compared with nonchinook days. Analysis of chinook wind velocities revealed that for chinook days, the relative risk of migraine onset was increased only on high-wind chinook days (velocity > 38 km/h) (odds ratio 1.41; 95% CI 1.06 to 1.88). A subset of individuals was sensitive to high-wind chinook days, and another subset was only sensitive to prechinook days. Only two patients were sensitive to both weather conditions, and the majority of patients was not sensitive to either. Neither weather condition had a protective effect. Increasing age was associated with high-wind chinook sensitivity (p = 0.009) but not prechinook sensitivity (p = 0.389). CONCLUSIONS: Both prechinook and high-wind chinook days increase the probability of migraine onset in a subset of migraineurs. Because few subjects were found to be sensitive to both weather types, the mechanisms for these weather effects may be independent. This is supported by the presence of an age interaction for high-wind chinook days but not for prechinooks day. PMID- 10668688 TI - Headache and major depression: is the association specific to migraine? AB - OBJECTIVE: To examine the relationship between migraine and major depression, by estimating the risk for first-onset major depression associated with prior migraine and the risk for first migraine associated with prior major depression. We also examined the extent to which comorbidity with major depression is specific to migraine or is observed in other severe headaches. METHODS: Representative samples of persons 25 to 55 years of age with migraine or other severe headaches (i.e., disabling headaches without migraine features) and controls with no history of severe headaches were identified by a telephone survey and later interviewed in person to ascertain history of common psychiatric disorders. RESULTS: Lifetime prevalence of major depression was approximately three times higher in persons with migraine and in persons with severe headaches compared with controls. Significant bidirectional relationships were observed between major depression and migraine, with migraine predicting first-onset depression and depression predicting first-onset migraine. In contrast, persons with severe headaches had a higher incidence of first-onset major depression (hazard ratio = 3.6), but major depression did not predict a significantly increased incidence of other severe headaches (hazard ratio = 1.6). CONCLUSIONS: The contrasting results regarding the relationship of major depression with migraine versus other severe headaches suggest that different causes may underlie the co-occurrence of major depression in persons with migraine compared with persons with other severe headaches. PMID- 10668689 TI - Chronic daily headache in Chinese elderly: prevalence, risk factors, and biannual follow-up. AB - OBJECTIVE: To investigate the prevalence, risk factors, and prognosis of chronic daily headache (CDH) in a population of elderly Chinese subjects. METHODS: A community-based survey of registered residents > or =65 years old (n = 2,003) in two townships of Kinmen Island in 1993. A neurologist used a structured questionnaire and clinical interview to make the diagnosis of headache. Subjects who had headaches > or =15 days/month for > or =6 months in the previous year were considered to have CDH. CDH was further classified into chronic tension-type headache (CTTH), CDH with migrainous features (CDH/MF), and other CDH. Person-to person biannual follow-up of the subjects with CDH was done in June 1995 and August 1997. RESULTS: A total of 1,533 people (77%) participated in our prevalence study. Sixty subjects (3.9%) fulfilled the criteria for CDH, with a higher prevalence in women (F/M: 5.6%/1.8%, p < 0.001). Of these subjects, 42 (70%) had CTTH, 15 (25%) had CDH/MF, and 3 (5%) had other CDH. Only 23% of those with CDH had consulted physicians for their headaches in the previous year. Multivariate logistic regression revealed the significant risk factors for CDH to be analgesic overuse (OR = 79), a history of migraine (OR = 6.6), and a Geriatric Depression Scale-Short Form score of > or =8 (OR = 2.6). The follow-up results in 1995 and 1997 showed that about two-thirds of the subjects still had CDH. Analgesic overuse (relative risk = 1.6) in 1993 was a significant predictor of persistent CDH at follow-up. CONCLUSIONS: A total of 3.9% of this elderly population had CDH, with CTTH being the most common subtype. Almost two-thirds of those with CDH had persistent frequent headaches at follow-up. Analgesic overuse was a significant predictor of a poor outcome. PMID- 10668690 TI - Idiopathic intracranial hypertension: relationship to depression, anxiety, and quality of life. AB - OBJECTIVE: To explore the incidence of depression and anxiety and to measure quality of life in women with idiopathic intracranial hypertension (IIH), a matched group cross-sectional study was conducted. Women with IIH (n = 28) were compared with control groups of weight- and age-matched women not diagnosed with IIH (n = 30) and with age-matched women of normal weight (n = 30). METHODS: Eighty-eight women completed a questionnaire soliciting health information and standardized questionnaires measuring depression, anxiety, and quality of life. The groups were compared using analysis of variance and chi2 tests. Where appropriate, post hoc comparisons were made using Fisher's test. RESULTS: Patients with IIH reported a greater number of adverse health problems than either of the control groups. Non-health-related psychosocial concerns were equally prevalent among the three groups, but IIH patients were significantly more affected by hardships associated with health problems than the other two groups. The patient group also had higher levels of depression and anxiety than the control groups. These adverse health conditions were reflected in decreased quality of life measures for the IIH patients. CONCLUSIONS: This study supports previous reports that link obesity and psychosocial difficulties, but obesity alone is not the explanation for the higher levels of depression and lower levels of quality of life. PMID- 10668691 TI - Ketone bodies do not directly alter excitatory or inhibitory hippocampal synaptic transmission. AB - OBJECTIVE: To determine the effect of the ketone bodies beta-hydroxybutyrate (betaHB) and acetoacetate (AA) on excitatory and inhibitory neurotransmission in the mammalian CNS. BACKGROUND: The ketogenic diet is presumed to be an effective anticonvulsant regimen for some children with medically intractable seizures. However, its mechanism of action remains a mystery. According to one hypothesis, ketone bodies have anticonvulsant properties. METHODS: The authors examined the effect of betaHB and AA on excitatory and inhibitory synaptic transmission in rat hippocampal-entorhinal cortex slices and cultured hippocampal neurons. In cultured neurons, their effect was also directly assayed on postsynaptic receptor properties. Finally, their ability to prevent spontaneous seizures was determined in a hippocampal-entorhinal cortex slice model. RESULTS: betaHB and AA did not alter synaptic transmission in these models. CONCLUSIONS: The anticonvulsant properties of the ketogenic diet do not result from a direct effect of ketone bodies on the primary voltage and ligand gated ion channels mediating excitatory or inhibitory neurotransmission in the hippocampus. PMID- 10668692 TI - 11C-flumazenil PET in patients with refractory temporal lobe epilepsy and normal MRI. AB - BACKGROUND: Using 11C-flumazenil (FMZ) PET with correction for partial-volume effect, reductions of central benzodiazepine receptor (cBZR) binding can be detected reliably in vivo on remaining neurons in sclerotic hippocampi of patients with mesial temporal lobe epilepsy (TLE). OBJECTIVE: To delineate abnormalities of 11C-FMZ binding in patients with medically refractory TLE and normal quantitative MRI. METHODS: Analysis of parametric images of FMZ volume of distribution (Vd) using two complementary approaches: 1) MRI-based volume of interest (VOI) approach with partial volume effect correction for multiple hippocampal and extrahippocampal VOIs; and 2) statistical parametric mapping (SPM) to localize significant 11C-FMZ binding changes objectively on a voxel-by voxel basis. RESULTS: Significant abnormalities of absolute FMZ-Vd were found after partial volume effect correction in 5 of 10 patients: unilateral decrease in the amygdala ipsilateral to the EEG focus (1), unilateral hippocampal decreases and bilateral temporal and extratemporal neocortical decreases (2), unilateral increase in the temporal neocortex together with extratemporal neocortical increases (1), and bilateral posterior hippocampal increases together with temporal neocortical increases (1). In the three patients with extratemporal neocortical changes, the concomitant unilateral hippocampal or temporal neocortical changes were contralateral to the presumed epileptic focus. Significant asymmetries of FMZ-Vd between homologous regions were found in six patients. In four of those patients, absolute FMZ-Vd for the homologous regions were within normal limits, with two of the four patients showing relatively higher hippocampal values ipsilateral to the presumed epileptic focus. SPM analysis localized significant abnormalities of FMZ-Vd in similar locations in three of the seven patients in whom VOI analysis detected significant changes. In addition, SPM indicated significant unilateral contralateral hippocampal decreases in an eighth patient. However, both methods failed to localize epileptic foci in two patients identified by depth-EEG recordings. CONCLUSIONS: 11C-FMZ PET showed focal increases as well as decreases of FMZ binding in 80% of patients with refractory TLE and normal high-quality MRI but was not consistently helpful in localizing the epileptic foci. PMID- 10668693 TI - Prevalence of nonconvulsive status epilepticus in comatose patients. AB - BACKGROUND: Nonconvulsive status epilepticus (NCSE) is a form of status epilepticus (SE) that is an often unrecognized cause of coma. OBJECTIVE: To evaluate the presence of NCSE in comatose patients with no clinical signs of seizure activity. METHODS: A total of 236 patients with coma and no overt clinical seizure activity were monitored with EEG as part of their coma evaluation. This study was conducted during our prospective evaluation of SE, where it has been validated that we identify over 95% of all SE cases at the Medical College of Virginia Hospitals. Only cases that were found to have no clinical signs of SE were included in this study. RESULTS: EEG demonstrated that 8% of these patients met the criteria for the diagnosis of NCSE. The study included an age range from 1 month to 87 years. CONCLUSION: This large-scale EEG evaluation of comatose patients without clinical signs of seizure activity found that NCSE is an underrecognized cause of coma, occurring in 8% of all comatose patients without signs of seizure activity. EEG should be included in the routine evaluation of comatose patients even if clinical seizure activity is not apparent. PMID- 10668694 TI - Discontinuation of antiepileptic drugs after successful epilepsy surgery. AB - OBJECTIVE: To evaluate the frequency and risk factors for seizure recurrence subsequent to antiepileptic drug (AED) withdrawal in patients who underwent surgical treatment for intractable partial epilepsy and were rendered seizure free. METHODS: The outcome of discontinuation of AED medication was studied retrospectively in 210 consecutive patients who were rendered seizure-free after epilepsy surgery performed between 1989 and 1993. RESULTS: Medical therapy was reduced in 96 patients and discontinued in 84 patients. The seizure recurrence rate after complete AED withdrawal was 14% and 36% at 2 and 5 years. In contrast, only 3% and 7% of the 30 patients who did not alter AED treatment after surgery had recurrent seizures in the same time intervals. After AED discontinuation, seizures tended to recur more often in patients with normal preoperative MRI studies compared with those with focal pathology. However, this difference did not reach significance. Intraoperative electrocorticography, extent of surgical resection, postoperative EEG, and seizure-free duration after surgery were not predictive of seizure outcome after AED withdrawal. CONCLUSIONS: AED withdrawal was associated with seizure recurrence in a significant portion of patients rendered seizure-free by epilepsy surgery. Patients with a normal preoperative MRI study showed a tendency for higher seizure recurrence, whereas the duration of seizure-free postoperative AED treatment interval did not significantly influence the seizure recurrence rate. These results will prove useful in counseling patients about discontinuing AED treatment after successful epilepsy surgery. PMID- 10668695 TI - Status epilepticus in stroke: report on a hospital-based stroke cohort. AB - OBJECTIVE: To evaluate occurrence rate, clinical data, and prognostic factors of status epilepticus (SE) after stroke. METHODS: From 1984 to 1994, 3,205 patients were admitted to the Department of Neurology at our institution with first-time strokes. A total of 159 of these patients had first-time poststroke seizures. Among these 159 patients, cases of SE were identified and evaluated. RESULTS: SE was recognized in 31 patients (19%). In 17 patients, SE was the first epileptic symptom (initial SE), and in 4 patients, stroke began with SE (S-SE). In the 14 remaining patients, SE occurred after one or more seizure(s). After a mean follow up period of 47 months, neurologic deterioration occurred after SE in 15 patients. This deterioration was permanent in two patients. Fifteen patients died; in five patients, death was directly related to SE. Eight of the 17 patients with initial SE and all 14 patients with SE after one or more seizure(s) developed other seizures or SE. S-SE, however, was not a predictive factor for additional seizure(s). CONCLUSIONS: Status epilepticus is common among patients with poststroke seizures. Although the immediate prognosis of patients with status epilepticus is poor, status epilepticus as the presenting sign did not necessarily predict subsequent epilepsy. PMID- 10668696 TI - Incidence and prognostic significance of fever following intracerebral hemorrhage. AB - OBJECTIVE: To investigate the incidence and prognostic significance of fever on presentation and during the subsequent 72 hours in patients with spontaneous supratentorial intracerebral hemorrhage (ICH). METHODS: We analyzed 251 patients. On admission, body temperature, Glasgow Coma Scale (GCS) score, age, sex, blood pressure, blood glucose level, and presumed origin of hemorrhage were analyzed. From the initial CT scan, hematoma volume, location, and presence of intraventricular hemorrhage were determined. From the first 72 hours, hematoma enlargement, duration of increased temperatures, blood pressure, and blood glucose level were determined. Outcome was classified on discharge with the Glasgow Outcome Scale (GOS) score. RESULTS: Outcomes included no symptoms in 23 (9%), moderate disability in 64 (26%), severe disability in 104 (41%), vegetative state in 5 (2%), and death in 55 (22%) patients. Prognostic factors retained from a logistic regression model with a dichotomized GOS scale (GOS score of 1 or 2 versus GOS score of 3 to 5) as response variables were GCS score of 7 or less, age older than 75 years, hematoma volume of more than 60 cm3, ventricular hemorrhage, and presence of a coagulation disorder (p < 0.05). Fever was associated with intraventricular hemorrhage. From 196 patients, data from the first 72 hours were analyzed. A total of 18 patients (9%) had normal temperatures throughout the study. The duration of fever (> or =37.5 degrees C) was less than 24 hours in 66 (34%), 24 to 48 hours in 70 (36%), and more than 48 hours in 42 patients (21%). Independent prognostic factors during the first 72 hours were duration of fever, secondary hemorrhage, GCS score of 7 or less, ventricular hemorrhage, hematoma volume of more than 60 cm3, duration of increased blood pressure of more than 48 hours, and duration of increased blood glucose of more than 48 hours. CONCLUSIONS: The incidence of fever after supratentorial ICH is high, especially in patients with ventricular hemorrhage. In patients surviving the first 72 hours after hospital admission, the duration of fever is associated with poor outcome and seems to be an independent prognostic factor in these patients. PMID- 10668697 TI - Hypoxia, hyperoxia, ischemia, and brain necrosis. AB - BACKGROUND: Human brains show widespread necrosis when death occurs after coma due to cardiac arrest, but not after hypoxic coma. It is unclear whether hypoxia alone can cause brain damage without ischemia. The relationship of blood oxygenation and vascular occlusion to brain necrosis is also incompletely defined. METHODS: We used physiologically monitored Wistar rats to explore the relationship among arterial blood oxygen levels, ischemia, and brain necrosis. Hypoxia alone (PaO2 = 25 mm Hg), even at a blood pressure (BP) of 30 mm Hg for 15 minutes, yielded no necrotic neurons. Ischemia alone (unilateral carotid ligation) caused necrosis in 4 of 12 rats, despite a PaO2 > 100 mm Hg. To reveal interactive effects of hypoxia and ischemia, groups were studied with finely graded levels of hypoxia at a fixed BP, and with controlled variation in BP at fixed PaO2. In separate series, focal ischemic stroke was mimicked with transient middle cerebral artery (MCA) occlusion, and the effect of low, normal, and high PaO2 was studied. RESULTS: Quantitated neuropathology worsened with every 10 mm Hg decrement in BP, but the effect of altering PaO2 by 10 mm Hg was not as great, nor as consistent. Autoradiographic study of cerebral blood flow with 14C iodoantipyrine revealed no hypoxic vasodilatation during ischemia. In the MCA occlusion model, milder hypoxia than in the first series (PaO2 = 46.5 +/- 1.4 mm Hg) exacerbated necrosis to 24.3 +/- 4.7% of the hemisphere from 16.6 +/- 7.0% with normoxia (PaO2 = 120.5 +/- 4.1 mm Hg), whereas hyperoxia (PaO2 = 213.9 +/- 5.8 mm Hg) mitigated hemispheric damage to 7.50 +/- 1.86%. Cortical damage was strikingly sensitive to arterial PaO2, being 12.8 +/- 3.1% of the hemisphere with hypoxia, 7.97 +/-4.63% with normoxia, and only 0.3 +/- 0.2% of the hemisphere with hyperoxia (p < 0.01), and necrosis being eliminated completely in 8 of 10 animals. CONCLUSIONS: Hypoxia without ischemia does not cause brain necrosis but hypoxia exacerbates ischemic necrosis. Hyperoxia potently mitigates brain damage in this MCA occlusion model, especially in neocortex. PMID- 10668698 TI - Stroke in children: the coexistence of multiple risk factors predicts poor outcome. AB - OBJECTIVE: To characterize the risk factors for stroke in children and their relationship to outcomes. METHODS: We reviewed charts of children with ischemic and hemorrhagic stroke seen at Hopital Sainte-Justine, Montreal between 1991 and 1997. RESULTS: We found 51 ischemic strokes: 46 arterial and 5 sinovenous thromboses. Risk factors were variable and multiple in 12 (24%) of the 51 ischemic strokes. Ischemic stroke recurred in 3 (8%) patients with a single or no identified risk factor and in 5 (42%) of 12 patients with multiple risk factors (p = 0.01). We also found 21 hemorrhagic strokes, 14 (67%) of which were caused by vascular abnormalities. No patient with hemorrhagic stroke had multiple risk factors. Hemorrhagic stroke recurred in two patients (10%). Outcome in all 72 stroke patients was as follows: asymptomatic, 36%; symptomatic epilepsy or persistent neurologic deficit, 45%; and death, 20%. Death occurred more frequently in patients with recurrent stroke (40%) than in those with nonrecurrent stroke (16%). CONCLUSIONS: Multiple risk factors are found in many ischemic strokes and may predict stroke recurrence. Recurrent stroke tends to increase rate of mortality. Because of the high prevalence and importance of multiple risk factors, a complete investigation, including hematologic and metabolic studies and angiography, should be considered in every child with ischemic stroke, even when a cause is known. PMID- 10668699 TI - Evaluation of antiretroviral drug efficacy for HIV-1 encephalitis in SCID mice. AB - OBJECTIVES: To compare the efficacy of the nucleoside reverse transcriptase inhibitors (NRTIs) abacavir, zidovudine (AZT), lamivudine (3TC), didanosine (ddI), and stavudine (d4T) to inhibit viral replication in brain macrophages. A severe combined immunodeficiency (SCID) mouse model of HIV-1 encephalitis (HIVE) was used to monitor spreading viral infection in the CNS. BACKGROUND: The development of antiretroviral therapies with CNS efficacy against neuroinvasive virus is important if eradication of HIV-1 can be achieved within critical "hidden reservoirs." METHODS: HIV-1-infected human monocyte-derived macrophages (MDMs) (after a single round of viral replication) were inoculated into the caudate and putamen of SCID mice. This resulted in the spreading of viral infection with a concomitant multinucleated giant cell encephalitis (astrogliosis, microglial activation, and neuronal injury). NRTIs were administered to animals at the time of intracerebral MDM inoculations and continued until the time of sacrifice. Antiretroviral effects were assessed by viral load and percentages of infected MDMs. RESULTS: In brains of SCID mice with HIVE, abacavir and lamivudine reduced HIV-1 p24 antigen-positive cells by 80% and 95%, respectively, whereas both decreased viral load by approximately 1 log. Zidovudine, didanosine, and stavudine showed variable effects. CONCLUSION: Abacavir and lamivudine showed significant antiretroviral activity in SCID mice with HIVE when compared with other NRTIs. The extrapolation of these results to humans with HIV-1 dementia awaits future investigations. PMID- 10668700 TI - Perfusion MRI detects rCBF abnormalities in early stages of HIV-cognitive motor complex. AB - OBJECTIVE: To evaluate patients with early HIV-cognitive motor complex (HIV-CMC) for possible regional cerebral blood flow (rCBF) abnormalities on perfusion MRI (pMRI). BACKGROUND: Nuclear medicine techniques have demonstrated global and focal cerebral perfusion abnormalities in patients with HIV dementia. Ultrafast pMRI enables the measurement of rCBF throughout the brain without the need to apply radioactive tracers or ionizing radiation. METHODS: pMRI was used to measure the rCBF in 19 patients with early stages of HIV-CMC and 15 healthy seronegative control subjects. The rCBF maps were registered to high-resolution anatomic MRI scans and transformed into Talairach space. Statistical analysis of the rCBF maps was performed with SPM96. RESULTS: Compared with the control subjects, the patients with HIV had statistically significantly decreased rCBF bilaterally in the inferior lateral frontal cortices (right: -15%, p < 0.002; left: -12%, p < 0.005) and in the inferior medial parietal brain region (-15%, p < 0.0009). In contrast, rCBF was increased bilaterally in the posterior inferior parietal white matter (right: +19%, p < 0.0001; left: + 17%, p < 0.001). Furthermore, rCBF abnormalities correlated significantly with clinical disease severity as measured by CD4 count, plasma viral load, Karnofsky score, and HIV dementia scale. DISCUSSION: Our results are consistent with previous findings from PET and SPECT studies. Furthermore, pMRI can detect rCBF abnormalities that correlate with disease severity in HIV-CMC. Because pMRI is more cost-effective, faster, and safer than nuclear medicine techniques for monitoring rCBF changes, pMRI may be more feasible for monitoring the effects of therapy for HIV-CMC. PMID- 10668701 TI - The association between APOE and dementia does not seem to be mediated by vascular factors. AB - OBJECTIVE: The effect of APOE on dementia may be mediated through dyslipidemia and atherogenesis through its effect on cholesterol metabolism. The authors investigated this possibility among aged survivors from the UK Medical Research Council Trial of the Treatment of Hypertension in Older Adults. DESIGN: A total of 370 of 657 survivors from an initial cohort of 1,088 recruited into the trial between 1983 and 1985 were traced in 1994 and agreed to be screened for dementia. Blood samples were analyzed for APOE genotype and serum fibrinogen. Cholesterol level, smoking behavior, blood pressure, body mass index, and EKG recordings had been measured at recruitment 10 to 12 years earlier. Odds ratios (ORs) for the association between APOE epsilon4/* and both AD and dementia were estimated and adjusted incrementally for the effect of age and premorbid intelligence, cholesterol, other risk factors for vascular disease, and EKG evidence of cardiovascular disease. RESULTS: The authors diagnosed 24 cases of National Institute of Neurological and Communicative Disorders and Stroke AD from 41 cases of dementia. The crude OR for the association between APOE epsilon4/* and AD was 3.40 (95% CI 1.30 to 8.91). APOE genotype was associated with serum cholesterol level, and there was a nonsignificant trend for an association with smoking behavior. After adjusting for these and all other vascular risk factors and vascular disease variables listed earlier, the OR for the association between APOE epsilon4/* and AD increased to 4.81 (1.60 to 14.4). CONCLUSION: Presence of APOE epsilon4/* seems to increase the risk for dementia and AD independently of its effect on dyslipidemia and atherogenesis. PMID- 10668702 TI - E4 allele dosage does not predict cholinergic activity or synapse loss in Alzheimer's disease. AB - OBJECTIVE: To investigate the relationship between apolipoprotein E (APOE) genotype and both cholinergic dysfunction and synapse loss in AD. BACKGROUND: A reduction in neocortical synapses and marked losses in the cholinergic system occur in AD. It has been suggested that the number of APOE epsilon4 alleles is inversely related to choline acetyltransferase (ChAT) activity, thereby influencing cholinergic function. Whether APOE genotype may influence neocortical synapse loss remains unclear. METHODS: An autopsy series of 182 patients with AD (National Institute on Aging and Consortium to Establish a Registry for Alzheimer's Disease criteria) and 16 normal controls (NC). APOE genotype was determined in blood samples or in postmortem brain tissue. Midfrontal synapse counts (AU/microg) were quantified by a dot-immunobinding assay for synaptophysin (Syn). Midfrontal ChAT activity (nmol/h/100 mg) was assessed using standard assays. RESULTS: Mean midfrontal ChAT activity and Syn were both significantly reduced in patients with AD compared with NC. The relationship between ChAT activity and number of epsilon4 allele copies in AD was complex, with ChAT activity lower in patients with either two or no epsilon4 alleles compared with those with one epsilon4 allele. There was no relationship between APOE genotype and synapse loss in AD. Syn density was almost identical across the three genotypes. CONCLUSIONS: Unlike other studies, we failed to detect a linear relationship between ChAT activity and number of epsilon4 allele copies in the midfrontal cortex of this large sample of patients with AD. Our data also show that the presence of epsilon4 allele does not influence midfrontal synapse loss in AD. This suggests that factors other than APOE genotype may be operative in the decline in midfrontal cholinergic function and synapses seen in AD. PMID- 10668703 TI - Cholinergic dysfunction in diseases with Lewy bodies. AB - OBJECTIVE: To evaluate cholinergic activity in diseases with Lewy bodies (LB; LB variant of AD [LBV], diffuse LB disease [DLBD], and Parkinson's disease [PD]) to determine if 1) AD changes are requisite to cholinergic dysfunction, 2) cholinergic activity declines to the same extent in neocortical and archicortical areas, and 3) cholinergic loss is influenced by APOE genotype. BACKGROUND: Like AD, diseases with LB are associated with decreased choline acetyltransferase (ChAT) activity. Increased APOE epsilon4 allele frequency has been reported in LBV. Whether APOE genotype affects cholinergic function in LBV remains unclear. METHODS: An autopsy series of 182 AD (National Institute on Aging and Consortium to Establish a Registry for Alzheimer's Disease criteria), 49 LBV, 11 PD, 6 DLBD, and 16 normal control (NC) subjects. APOE genotype and ChAT activity (nmol/h/100 mg) in the midfrontal and hippocampal cortices were determined. RESULTS: Mean midfrontal ChAT activity was markedly reduced in diseases with LB (LBV: 53.3 +/- 39.0; PD: 54.8 +/- 35.7; DLBD: 41.3 +/- 24.8) compared to NC (255.4 +/- 134.6; p < 0.001) and AD (122.6 +/- 78.9; p < 0.05). Among diseases with LB, midfrontal ChAT activity was decreased to a similar extent in patients with (LBV) and without (DLBD and PD) AD pathology. Although mean ChAT activity for LBV was less than half that for AD in the midfrontal cortex, it was similar to that for AD in the hippocampus (LBV: 243.5 +/- 189.7; AD: 322.8 +/- 265.6; p > 0.05). However, hippocampal ChAT activity for both AD and LBV was lower than that for NC (666.5 +/- 360.3; p < 0.001). The epsilon4 allele dosage did not influence midfrontal ChAT activity in LBV. CONCLUSION: Marked losses in midfrontal ChAT activity occur in diseases with LB, independent of coexistent AD changes. A greater midfrontal, as opposed to hippocampal, cholinergic deficit may differentiate LBV from AD. The lack of a relationship between epsilon4 allele dosage and midfrontal ChAT activity suggests that other factors may play a role in its decline in LBV. PMID- 10668704 TI - APOE epsilon4 does not predict mortality, cognitive decline, or dementia in the oldest old. AB - OBJECTIVE: To examine the effect of the epsilon 4 allele on cognitive decline in the oldest old. METHODS: We studied all 601 citizens of the city of Vantaa age 85 years and older in 1991. A total of 553 subjects (92%) took part in the study, which used the Mini-Mental State Examination (MMSE) and assessment of dementia according to the Diagnostic and Statistical Manual of Mental Disorders, third ed., revised (DSM-III-R) criteria. The survivors were re-examined 3 years later. APOE genotype was determined in 510 subjects, representing 83.2% of the original population. RESULTS: Approximately one-half of the subjects (n = 250) died before the follow-up, and 253 subjects (97.3% of the survivors) were re-examined. The occurrence of the APOE epsilon 4 allele did not have any significant effect on survival. Of the 187 previously nondemented subjects, 58 (31%) had developed dementia. The OR for the epsilon 4 carriers to develop dementia was not significant: OR = 1.78; 95% CI = 0.88 to 3.60. In individuals with a follow-up MMSE score (n = 222), the mean decline in the score was 3.1 points. APOE epsilon 4 carrier status did not have a significant effect on the mean MMSE change except in the previously demented subjects, among whom the drop was larger in the APOE epsilon 4 carriers. CONCLUSIONS: The lack of association between APOE epsilon 4 carrier status and mortality, or development of dementia, or cognitive decline in these very elderly people, whether analyzed in the whole population or among the nondemented subjects only, suggests that the APOE epsilon 4 effect in younger subjects is age-dependent, and that it is no longer present in very old age. PMID- 10668705 TI - Early-life risk factors and the development of Alzheimer's disease. AB - OBJECTIVE: To investigate the association of early-life factors with AD. BACKGROUND: The early-life environment and its effect on growth and maturation of children and adolescents are linked to many adult chronic diseases (heart disease, stroke, hypertension, and diabetes mellitus), and these effects are also linked to maternal reproduction. AD may have an early-life link. The areas of the brain that show the earliest signs of AD are the same areas of the brain that take the longest to mature during childhood and adolescence. A poor-quality childhood or adolescent environment could prevent the brain from reaching complete levels of maturation. Lower levels of brain maturation may put people at higher risk for AD. METHODS: In a community-based case-control study (393 cases, 377 controls), we investigated the association of early-life factors and AD. Early-life variables include mother's age at patient's birth, birth order, number of siblings, and area of residence before age 18 years. Patient education level and apolipoprotein E (APOE) genotypes were also included in the analysis. RESULTS: Area of residence before age 18 years and number of siblings are associated with subsequent development of AD. For each additional child in the family the risk of AD increases by 8% (OR = 1.08, 95% CI = 1.01 to 1.15). More controls compared with cases grew up in the suburbs (OR = 0.45, 95% CI = 0.25 to 0.82). APOE epsilon 4 and the patient's education level did not confound or modify the associations. CONCLUSIONS: The early-life childhood and adolescent environment is associated with the risk of AD. PMID- 10668706 TI - Lu 25-109, a muscarinic agonist, fails to improve cognition in Alzheimer's disease. Lu25-109 Study Group. AB - OBJECTIVE: To evaluate the therapeutic effect of the selective muscarinic receptor m1 partial agonist, m2 antagonist, Lu25-109-a compound that directly stimulates muscarinic cholinergic receptors-in patients with probable AD. METHODS: A 6-month, randomized, double-blind, placebo-controlled, parallel group trial comparing three doses of Lu25-109 with placebo was carried out. A total of 496 patients with probable AD with a Mini-Mental State Examination score between 10 and 26 were enrolled at 29 centers and randomized to placebo or Lu25-109 25, 50, or 100 mg tid. The primary efficacy measures were the AD Assessment Scale Cognitive subscale and the AD Cooperative Study Clinical Global Impression of Change. Secondary efficacy variables included the AD Cooperative Study Inventory of Activities of Daily Living and the Behavioral Symptoms in AD Scale. RESULTS: In both an intent-to-treat and a completer's analysis there were no significant differences for either the two primary or the secondary variables. There was a trend for patients on the highest drug dose to worsen in the completer's analysis. Adverse events included dizziness, nausea, diarrhea, fatigue, increased sweating, and anorexia, all of which increased with increasing drug dose. CONCLUSION: Lu25-109, a selective partial ml agonist and an m2/m3 antagonist, failed to improve cognition in patients with mild to moderate AD. PMID- 10668707 TI - Gender differences in the treatment of behavior problems in Alzheimer's disease. SAGE Study Group. Systemic Assessment of Geriatric drug use via Epidemiology. AB - OBJECTIVE: To define gender differences in noncognitive behavioral problems of patients with AD and differences in the associated treatment of those problems. DESIGN/METHODS: We performed an observational study using the Systematic Assessment and Geriatric drug use via Epidemiology (SAGE) database, which contains data collected with the Minimum Data Set on a cross-section of nursing home residents in five US states. Behavior problems were documented at the first assessment of 28,367 residents with AD. We evaluated the role of gender differences in behavior as predictors of differences in nonpharmacologic versus specific pharmacologic therapies with psychoactive medications using logistic regression. RESULTS: Men were more likely than women to exhibit behavior problems such as wandering, abusiveness, and social impropriety (59% versus 50% for any behavior problem). Hallucinations and delusions as well as depression were equally prevalent in men and women. Nevertheless, men were more likely to receive psychoactive medications. Among the specific drug categories examined, and controlling for age and degree of cognitive impairment, men were more likely to receive antipsychotic drugs and less likely to be receiving antidepressants. CONCLUSION: Gender appears to play an important role in determining the frequency of behavioral problems in nursing home residents with AD, which may influence choice of treatments as well as the decision whether to treat. The use of more potent tranquilizers in men with problem behaviors has potential implications for morbidity, deserving further investigation. PMID- 10668708 TI - Lack of association of the alpha2-macroglobulin locus on chromosome 12 in AD. AB - OBJECTIVE: Analysis of AD has revealed that the apolipoprotein E locus (APOE) cannot account for all of the genetic risk associated with AD. Whole genome scanning in AD families suggests that a chromosome 12 locus may contribute significantly to disease development. The alpha2-macroglobulin gene (A2M) has been suggested as a candidate locus for AD based on analysis of familial AD. METHOD: We determined, in 195 neuropathologically verified AD cases and 107 age matched control subjects, the association of two common polymorphisms in A2M (a pentanucleotide deletion 5' to the bait domain exon, and a valine-1000-isoleucine polymorphism in the thiolester site of the protein). RESULTS: Evidence was observed for linkage disequilibrium between the deletion and Ile1000 polymorphisms. No evidence was observed for an association between the thiolester polymorphism and AD alone or when accounting for the APOE-epsilon4 allele. No alteration in the frequency of the bait domain deletion was observed, although a small excess (4%) of deletion homozygotes was found in the AD group, which were absent in the control population. CONCLUSIONS: The A2M deletion polymorphism at most accounts for a small fraction of the genetic contribution toward AD, and this is small compared with APOE. Furthermore, reverse transcriptase PCR of A2M RNA from the brains of patients homozygous for the deletion polymorphism showed that the bait domain exon still is present in the RNA. This suggests that the A2M deletion polymorphism may be nonfunctional and that the chromosome 12 AD locus is situated elsewhere. PMID- 10668709 TI - Alpha2 macroglobulin and the risk of Alzheimer's disease. AB - BACKGROUND: alpha2 Macroglobulin is a panproteinase inhibitor that is found immunohistochemically in neuritic plaques, a requisite neuropathologic feature of AD. Recently, a pentanucleotide deletion near the 5' end of the "bait region" of the alpha2 macroglobulin (A2M) gene was reported to be associated with AD in a large cohort of sibpairs, in which the mutation conferred a similar odds ratio with AD as the APOE-epsilon4 allele for carriers of at least one copy of the A2M gene (Mantel-Haenszel odds ratio, 3.56). METHODS: We studied three independent association samples of AD patients (n = 309) with an age range of 50 to 94 years and representative controls (n = 281) to characterize the allele frequency of the pentanucleotide deletion in this cohort. We detected the mutation near the 5' splice site of exon 18 using standard PCR and restriction fragment length polymorphism methods. The results were adjusted for age, gender, education, and APOE polymorphism. RESULTS: We found that the A2M gene polymorphism conferred an increased risk for AD, with an estimated Mantel-Haenszel ratio of 1.5 (95% CI 1.1 to 2.2; p = 0.025). There was no age- or gender-dependent increase in A2M gene allele frequencies in AD patients compared with controls. The combined sample showed the expected association between AD and APOE-epsilon 4. In one of our three samples there was an interaction between the A2M and APOE-epsilon4 genes, but the other two samples showed no interaction between the two risk factors. CONCLUSIONS: Our data support an association between the A2M gene and AD. This association is less pronounced, however, in our cohort than in the previously reported sample of sibpairs. PMID- 10668710 TI - Internal carotid artery dissection. PMID- 10668711 TI - Alpha2-macroglobulin polymorphism is not associated with AD or AD-type neuropathology in the Japanese. AB - BACKGROUND: alpha2-Macroglobulin (A2M) forms the complex with amyloid beta protein (Abeta) and is associated with degradation of Abeta. It has been reported that the A2M gene (A2M) exon 18 splice acceptor deletion polymorphism influences the development of AD, regardless of apolipoprotein E-epsilon4 (APOE-epsilon4) status. OBJECTIVE: To determine the effect of A2M polymorphism on the development of AD and AD-type neuropathologic changes. METHODS: The authors examined the A2M and APOE genotypes, the densities of the senile plaques (SPs), SPs with dystrophic neurites (NPs), and neurofibrillary tangles (NFTs) in the brains of 62 postmortem-confirmed sporadic AD and 90 nondemented patients from an autopsy series of elderly Japanese subjects. RESULTS: There was no association of the A2M polymorphism with AD, age at onset, or duration of illness in AD. The A2M polymorphism was not associated with the SPs, NPs, or NFTs in AD or nondemented patients. The results remained insignificant, even when the A2M genotype groups were divided into subgroups by APOE-epsilon4 status. CONCLUSION: The A2M polymorphism does not affect the development of sporadic AD or formation of AD type neuropathologic changes. PMID- 10668712 TI - Prevalence and outcomes of vascular cognitive impairment. Vascular Cognitive Impairment Investigators of the Canadian Study of Health and Aging. AB - OBJECTIVE: To assess the importance of vascular cognitive impairment and its three subgroups (cognitive impairment, no dementia; vascular dementia; and AD with a vascular component) to the prevalence and burden of cognitive impairment in elderly people. BACKGROUND: Vascular lesions may produce a spectrum of cognitive changes. Omitting elderly patients whose cognitive impairment falls short of dementia (vascular cognitive impairment, no dementia) may give a falsely low indication of the prevalence and burden of disease. To test this proposition, we compared the rates of adverse outcomes for patients with no cognitive impairment, vascular cognitive impairment (and its subgroups), and probable AD. METHODS: The Canadian Study of Health and Aging is a prospective cohort study of 10,253 randomly selected community-dwelling and institution-dwelling respondents aged 65 years or older. In the community, all participants (n = 9,008) were screened for cognitive impairment; those who screened positive and a sample of those who screened negative received a clinical assessment (n = 1,659). All patients living in institutions received a clinical assessment (n = 1,255). Participants were reassessed 5 years after the original survey. RESULTS: Vascular cognitive impairment without dementia was the most prevalent form of vascular cognitive impairment among those aged 65 to 84 years. Rates of institutionalization and mortality for those with vascular cognitive impairment were significantly higher than those of people who had no cognitive impairment, and the mortality rate for patients with vascular cognitive impairment was similar to that of patients with AD. CONCLUSIONS: Failure to consider vascular cognitive impairment without dementia underestimates the prevalence of impairment and the risk for adverse outcomes associated with vascular cognitive impairment. PMID- 10668713 TI - Rate of functional decline in Huntington's disease. Huntington Study Group. AB - OBJECTIVE: To determine the rate of functional decline in a large cohort of patients with Huntington's disease (HD) followed at 43 sites by the Huntington Study Group (HSG). METHODS: The annual rate of functional decline was measured using the Total Functional Capacity Scale (TFC) and the Independence Scale (IS) in 960 patients with definite HD followed prospectively for a mean of 18.3 months. All patients were rated with the Unified Huntington's Disease Rating Scale (UHDRS). Sample size calculations for hypothetical clinical trials were calculated. RESULTS: A factor analysis of the UHDRS at baseline yielded 15 factors accounting for 77% of the variance. The TFC score declined at a rate of 0.72 units/year (standard error [SE] 0.04) and the IS score declined at a rate of 4.52 units/year (SE 0.23). Lower TFC score at baseline, indicating more severe impairment, was associated with less rapid annual decline in TFC score, perhaps reflecting the floor effect of the scale. The annual rate of decline for 575 patients with baseline TFC scores of 7 to 13 was 0.97 (SE 0.06), was 0.38 (SE 0.08) for 270 patients with baseline TFC scores of 3 to 6, and was 0.06 (SE 0.1) for 101 patients with TFC scores of 0 to 2. In multivariate analysis (n = 960), longer disease duration and better cognitive status at baseline were associated with a less rapid rate of decline in TFC score, whereas depressive symptomatology was the only factor associated with more rapid decline on the IS score. Age at onset of HD, sex, weight, and education did not affect decline on either score. CONCLUSIONS: The comparable rates of decline on the TFC and the IS scores with other published studies suggest that these estimates of functional decline are representative of HD patients who are evaluated at HSG research sites. In longitudinal analysis, longer disease duration and better neuropsychological performance at baseline were associated with a less rapid rate of decline in TFC score, whereas depressive symptomatology at baseline was associated with a more rapid decline in the IS score. These rates of functional decline and the covariates that modify them should be considered in estimating statistical power and designing future therapeutic trials involving HD patients with early or moderately severe disease. PMID- 10668714 TI - Effects of central dopaminergic stimulation by apomorphine on speech in Parkinson's disease. AB - OBJECTIVE: To determine the effect of central dopaminergic stimulation with apomorphine on speech in PD. BACKGROUND: Most patients with PD have a speech disorder. Of those, 89% have involvement of laryngeal function, and 45% have additional articulatory dysfunction. The effect of dopaminergic medications on these two dimensions of speech impairment in PD has not been selectively studied. METHODS: In a randomized, double-blind, placebo-controlled crossover design, patients with PD and speech impairment, Hoehn and Yahr stages 2 to 4 "off," and without severe dyskinesias were given placebo or apomorphine injections 0.05 mg/kg subcutaneously during two consecutive outpatient visits. They were pretreated with domperidone for 48 hours and were tested off their parkinsonian medications for 12 hours. Laryngeal function was assessed by maximum sustained vowel phonations and comfortable vowel phonations. Articulatory function was evaluated by speech intelligibility score, speaking rate, and efficiency ratio. RESULTS: Ten patients, mean age 73.4 years (SD = 6.6), disease duration 8.7 years (SD = 6.3), were tested. The baseline motor score on the Unified Parkinson's Disease Rating Scale (UPDRSm) and all experimental speech variables were equivalent on both placebo and apomorphine days. At a dose of apomorphine that provoked improvement in UPDRSm (p = 0.0078), no index of either laryngeal or articulatory function improved significantly after apomorphine administration. CONCLUSION: Laryngeal and articulatory speech components are not under prominent dopaminergic control in PD. Treatment regimens should focus on nondopaminergic pharmacology and other therapies. PMID- 10668715 TI - Autosomal dominant diffuse leukoencephalopathy with neuroaxonal spheroids. AB - OBJECTIVE: To provide clinical, MRI, and histopathologic findings in a rare white matter disorder with autosomal dominant inheritance, so-called hereditary diffuse leukoencephalopathy with spheroids (HDLS). BACKGROUND: Progressive leukoencephalopathies often constitute a diagnostic dilemma in both children and adults. In some cases, histopathologic examination of brain tissue is required for a classifying diagnosis. METHODS: Clinical history, MRI, and autopsy findings were reviewed in three patients with HDLS: a father, his daughter, and an unrelated patient. RESULTS: Clinical history consisted of an adult-onset neurologic deterioration with signs of frontal lobe dysfunction, epilepsy, spasticity, ataxia, and mild extrapyramidal disturbances. MRI findings included cerebral atrophy and patchy white matter changes, most pronounced in the frontal and frontoparietal area with extension through the posterior limb of the internal capsule into the pyramidal tracts of the brainstem. Autopsy in two patients revealed a leukoencephalopathy with frontoparietal and frontal preponderance and numerous neuroaxonal spheroids in the abnormal white matter. The pyramidal tracts were affected throughout the brainstem. CONCLUSION: Similar clinical and histopathologic findings have been reported in members of a Swedish pedigree. The homogeneity of the findings strongly suggests that HDLS is a distinct disease entity. In the absence of a biochemical or genetic marker, a definitive diagnosis requires histopathologic confirmation in one of the affected family members. Neuroaxonal spheroids. PMID- 10668716 TI - A randomized controlled trial of recombinant interferon beta-1a in ALS. Italian Amyotrophic Lateral Sclerosis Study Group. AB - OBJECTIVE: To evaluate the efficacy of recombinant interferon beta (IFNbeta)-1a in the treatment of ALS. BACKGROUND: It has been proposed that IFNs affect the progression of ALS by interfering with putative immune mechanisms involved in the pathogenesis of the disease. METHODS: Patients (n = 61) 40 to 70 years of age with a 6- to 24-month history of confirmed ALS with mild to moderate disability received IFNbeta-1a, 12 mIU (n = 31), or placebo (n = 30) subcutaneously three times a week for 6 months and were followed up for an additional 6 months. Patients were assessed after 4, 12, 24, 36, and 48 weeks. Medical Research Council scale, Norris scale, and bulbar scores as well as forced vital capacity were used to assess disability. Selected electrophysiologic measures (latency, amplitude, and duration of the compound muscle action potential) were also used. RESULTS: Twenty patients randomized to IFNbeta-1a and 17 patients given placebo completed the study. A total of 16 patients receiving IFNbeta-1a became non-self supporting compared with 16 on placebo (52% versus 53%). There were no significant differences between the two treatment groups for any of the measures of disease progression and disability. Deaths were reported in six patients treated with IFNbeta-1a and four patients on placebo. Adverse events were reported more frequently with IFNbeta-1a (77% of patients) compared with placebo (57%), with flu-like symptoms and local erythema being the commonest complaints. CONCLUSIONS: This pilot study suggests that IFNbeta-1a is not effective in the treatment of ALS. PMID- 10668717 TI - Mechanism of block of nicotinic acetylcholine receptor channels by purified IgG from seropositive patients with myasthenia gravis. AB - OBJECTIVE: To clarify the mechanism of block of nicotinic receptor channels by myasthenic antibodies. BACKGROUND: Nicotinic acetylcholine receptor (nAChR) channel currents are functionally blocked by purified immunoglobulin G (IgG) of patients with myasthenia gravis (MG). METHODS: The molecular mechanism of block of IgG fractions containing antibodies to nAChR channels was tested with the patch-clamp technique in combination with a system for ultrafast solution exchange. For the experiments, outside-out patches from cultured mouse myotubes that express embryonic-type nAChR channels at their surface were used. RESULTS: Incubation of outside-out patches with purified IgG from four myasthenic patients blocked nAChR channel currents activated by the application of 1.0 mM ACh reversibly. The peak current amplitude and the time course of block of nAChR channels decreased with increasing concentrations of IgG. The block became at least partly irreversible if incubation time of outside-out patches exceeded 2 minutes. For the block of nAChR channel currents with a-bungarotoxin, a similar mechanism of block was found. CONCLUSIONS: The reversibility of functional block of nAChR channel currents by myasthenic IgG depended strongly on the incubation time of the receptors with antibodies. Interaction of myasthenic antibodies with nicotinic receptors may proceed in several stages from a low-affinity reversible to a high-affinity irreversible binding. PMID- 10668718 TI - Training the future neurology workforce. AB - OBJECTIVE: To address training demands on future neurologists, the American Academy of Neurology (AAN) surveyed its US members as to their views about training the future neurology workforce. METHODS: The survey was mailed to 575 US neurologists and 425 residents/fellows. Respondents (54%) were asked about their perceptions of current and future educational programs and settings needed to improve practice competence; issues related to subspecialization; and the role of non-neurologists in providing neurologic care. Views of neurologists were compared with those of neurology residents/fellows. RESULTS: Most respondents support additional training in outpatient, community, and staff model health maintenance organization settings. The majority of respondents oppose a required fifth year of training or a yearly competency examination, but neurologists who have a subspecialty interest and residents/fellows favor elective certification and higher fees by subspecialists. General neurologists oppose these ideas. Most neurologists feel that primary care physicians, nurse practitioners, and physician assistants can manage uncomplicated neurologic problems, although residents/fellows are less willing to accept the role of nonphysician providers for neurologic services. CONCLUSIONS: Neurology educational programs should consider addressing deficiencies that today's practitioners perceive. Increasing subspecialization, although favored by most neurologists, creates a challenge for the neurologic community as neurologists without subspecialty training see this trend as a threat to their livelihood. PMID- 10668719 TI - No induction of apoptosis by IFN-beta in human antigen-specific T cells. AB - Interferon (IFN)-beta, the most effective immunomodulatory treatment for MS, inhibits the proliferation of myelin-specific T cells. We report that IFN-beta moderately enhances the expression of the death receptor, CD95, at the surface of human antigen-specific T cells. However, T-cell apoptosis was not induced by IFNbeta-1a or IFNbeta-1b as assessed by caspase activity or DNA fragmentation. Immunomodulation mediated by IFN-beta does not directly involve apoptotic pathways in human T cells. PMID- 10668720 TI - Identification of new and common mutations in the EPM2A gene in Lafora disease. AB - Lafora disease is a teenage onset progressive myoclonus epilepsy caused by mutations in the EPM2A gene. In this report, we describe new mutations within EPM2A, review the known mutations to date to identify the most common, and describe three simple tests for prenatal and carrier screening. PMID- 10668721 TI - An interrupted 34-CAG repeat SCA-2 allele in patients with sporadic spinocerebellar ataxia. AB - In spinocerebellar ataxia type 2 (SCA-2), a difference of three CAG repeats distinguishes normal alleles (14 to 31 repeats) from pathogenic alleles (34 to 57 repeats). All sequenced pathogenic alleles have a pure CAG repeat structure, whereas interrupted repeats have been seen exclusively in normal alleles. The authors present two patients with sporadic SCA with an interrupted 34-CAG repeat allele, (CAG)24(CAA)(CAG)9, who showed a phenotype compatible with SCA-2. The interrupted allele coding for a 34 pure polyglutamine tract may cause the SCA phenotype. PMID- 10668722 TI - Isolated musculocutaneous neuropathy caused by a proximal humeral exostosis. AB - We report an isolated musculocutaneous neuropathy caused by a proximal humeral osteochondroma that became symptomatic after the patient played recreational basketball. Lesion resection resulted in complete deficit resolution. Mass lesions involving the musculocutaneous nerve should be considered in patients with atraumatic, isolated musculocutaneous neuropathies that are recurrent or fail to recover, even in the setting of strenuous exercise. PMID- 10668723 TI - Atypical Friedreich ataxia phenotype associated with a novel missense mutation in the X25 gene. AB - We describe two sisters with early onset gait ataxia, rapid disease progression, absent or very mild dysarthria and upper limb dysmetria, retained knee jerks in one, slight to moderate peripheral nerve involvement, and diabetes. Molecular analysis showed that they are compound heterozygotes for GAA expansion and a novel exon 5a missense mutation (R165P). This mutation appears to be associated with an atypical but not milder Friedreich ataxia phenotype. PMID- 10668724 TI - Micturitional disturbance in pure autonomic failure. AB - We obtained micturitional histories and performed urodynamic studies in six patients with pure autonomic failure. All patients had urinary symptoms. Urodynamic studies showed postmicturition residuals in two, small bladder capacities in two, detrusor hyperreflexia in four, low bladder compliance in two, detrusor-external sphincter dyssynergia in one, neurogenic sphincter electromyography in three, and denervation supersensitivity of the bladder in two. Micturitional disturbance is a common feature in pure autonomic failure because of peripheral and central types of abnormalities. PMID- 10668725 TI - An FDOPA PET study in patients with periodic limb movement disorder and restless legs syndrome. AB - The authors investigated nine drug-naive patients with periodic limb movement disorder and restless legs syndrome (PLMD-RLS) and 27 healthy controls with PET using 6-[18F]fluoro-L-dopa (FDOPA). In the patients, the FDOPA uptake (Ki(occ)) in the caudate nucleus was 88% and in the putamen 89% of the control mean values. This equal affection of the caudate and the putamen differs, for example, from the dopaminergic dysfunction in Parkinson's disease, which affects the putamen earlier and more severely than the caudate. The current results indicate mild nigrostriatal presynaptic dopaminergic hypofunction in PLMD-RLS. PMID- 10668726 TI - A kindred with Parkinson's disease not showing genetic linkage to established loci. AB - We describe a kindred with PD with an onset age from the fifth to the eighth decade. Genetic analysis indicated that the genetic defect in this family was unlikely to be in the alpha-synuclein, parkin, or tau genes, or to reside on chromosomes 2p or 4p. PMID- 10668727 TI - A magnetization transfer imaging study of the brain in patients with migraine. AB - The authors evaluated the magnetization transfer ratio (MTR) of T2 lesions, normal-appearing white matter (NAWM), and brain from 39 migraineurs, 17 healthy volunteers, and 22 patients with MS. Migraineurs had NAWM and brain MTR values similar to those of normal subjects but significantly higher than those of MS patients. Average lesion MTR values also were significantly lower in MS patients than in migraineurs. In patients with migraine, other etiologies should be considered in the presence of tissue damage beyond that seen on T2-weighted scans. PMID- 10668728 TI - Decreased hemispheric water mobility in hemiplegic migraine related to mutation of CACNA1A gene. AB - We report a reversible reduction of water diffusion in the brain during a prolonged attack of hemiplegic migraine. The patient had a sporadic mutation of the CACNA1A gene. The diffusion changes were observed in the contralateral hemisphere 3 and 5 weeks after the onset of hemiplegia. These results suggest the occurrence of hemispheric cytotoxic edema during severe attacks of hemiplegic aura. The mechanisms underlying such ultrastructural modifications are unknown but an abnormal release of excitatory amino acids can be hypothesized. PMID- 10668729 TI - Ischemic strokes are more severe in Poland than in the United States. AB - Case fatality rates for stroke were ascertained prospectively in two regional catchment hospitals in Poland and 36 teaching hospitals in the US University Hospital Consortium. Case fatality rates in Poland (23.9%) were higher than in the United States (7.5%). Angina, atrial fibrillation, and congestive heart failure were more frequent in Polish stroke patients (40%, 26%, and 25%, respectively) than in US patients (17%, 12%, and 10%). Stroke severity as indicated by higher frequencies of hemiplegia, disordered consciousness, dysphagia, and aphasia was greater in Poland (19%, 39%, 28%, and 42%, respectively) than the United States (11%, 13%, 14%, and 26%). PMID- 10668730 TI - Exploratory saccades show no direction-specific deficit in neglect. AB - In patients with spatial neglect, contralesional reflexive saccades toward suddenly appearing targets show direction-specific deficits. We examined whether these deficits also occur during free exploration of space. Neglect patients' voluntary eye movements showed reduced amplitudes for saccades in all directions but no direction-specific deficit. The results argue against an interpretation of spatial neglect as a general deficit to disengage attention or to program saccades in contralesional direction. PMID- 10668731 TI - Spinal cord astrocytoma: response to PCV chemotherapy. AB - Information regarding the value of chemotherapy for spinal cord astrocytomas that progress after irradiation is limited. We describe a patient whose conus medullaris astrocytoma responded to PCV (procarbazine, lomustine, and vincristine) chemotherapy after failing radiation and cisplatin-based chemotherapy. PCV should be considered in patients with progressive spinal cord astrocytomas. PMID- 10668732 TI - Loss of ability to sneeze in lateral medullary syndrome. AB - Four consecutive patients with lateral medullary syndrome reported reversible inability to complete a reflex sneeze, despite retaining the urge to do so and the ability to mimic the motor act. This previously undescribed feature of a relatively common syndrome is in keeping with the known location of a "sneeze center" in the lateral medulla of cat. In man, unilateral brainstem lesion is sufficient to abolish the sneeze reflex temporarily. PMID- 10668733 TI - Right frontal areas 6 and 8 are associated with simultanapraxia, a subset of motor impersistence. AB - The authors examined brain lesions that cause simultanapraxia, a subset of motor impersistence. Simultanapraxia was defined as the inability to perform two motor acts simultaneously: closing the eyes and protruding the tongue. Simultanapraxia was found in 9 (5.6%) of 160 hospitalized patients with cerebrovascular diseases. The lesions were located in areas 6 and 8 in the right middle cerebral artery territory. This site was spared in five patients who did not show simultanapraxia, even with a large infarction in the right middle cerebral artery area. PMID- 10668734 TI - Hemodynamic changes in simple partial epilepsy: a functional MRI study. AB - We performed functional MRI (fMRI) on a patient with a mass lesion while she happened to experience a simple partial seizure. We used regional T2* signal changes to localize seizure-related hemodynamic changes. Seizure activity was associated with changes in MR signal in different regions that showed sequential activation and deactivation. Our study has shown that epileptic activity leads to changes in cerebral hemodynamics. In selected patients, therefore, it might be possible to use fMRI as a noninvasive tool to detect and investigate cortical patterns of activation associated with seizure activity. PMID- 10668735 TI - Spinal schistosomiasis. PMID- 10668736 TI - Selective sparing of pain pathways in a patient with adult cerebral adrenoleukodystrophy. PMID- 10668737 TI - Dystrophinopathy expressing as either cardiomyopathy or Becker dystrophy in the same family. PMID- 10668738 TI - Pupillary diameter assessment: need for a graded scale. PMID- 10668739 TI - A clinical examination technique for mild upper motor neuron paresis of the arm. PMID- 10668740 TI - Superior sagittal sinus thrombosis due to lithium: local urokinase thrombolysis treatment. PMID- 10668741 TI - Paroxysmal word deafness secondary to focal epilepsy. PMID- 10668742 TI - Efficacy of levodopa therapy on motor function after posteroventral pallidotomy for Parkinson's disease. PMID- 10668743 TI - A tentative interpretation of electromyographic regional differences in bulbar and limb-onset ALS. PMID- 10668744 TI - Creatine monohydrate increases strength in patients with neuromuscular disease. PMID- 10668745 TI - The biochemical pathway of neurofibrillary degeneration in aging and Alzheimer's disease. PMID- 10668746 TI - Bilateral infarction in the territory of the anterior cerebral arteries. PMID- 10668747 TI - Use of microvascular free-tissue transfer following ablative surgery of the skull base. AB - The purpose of this study was to validate the use of free flaps in reconstruction of skullbase defects after extensive resection of advanced tumors, and to justify microvascular reconstruction to improve the quality of life and survival in this population. The treatment outcome after ablative resection of skullbase tumors with free flap reconstruction over a 7-year period (1988 to 1995) was studied. Complete removal of the tumor was originally attempted in all patients. All cases had immediate reconstruction. Criteria for reconstruction with free flaps were based on extensive defects in which local flaps were insufficient. Twenty patients were identified male:female, 11:9). The most common tumor was sarcoma, followed by squamous-cell carcinoma. Coverage of the dura was required in 12 patients. Muscles used were the rectus abdominis and latissimus dorsi. Complications included flap necrosis (n = 2) and ventral hernia (n = 2). Control of pain was achieved in 66 percent of cases. Patients with regional metastasis died within 2 years, and those with distant metastasis died within 18 months. Patients with primary tumors had an increased survival rate. The authors confirm the technical feasibility and success of free flaps to reconstruct extensive defects in the skull base. In patients with potentially complete resection of primary/recurrent lesions, overall survival justifies the procedure. Patients with regional/distant metastasis warrant an individualized approach. PMID- 10668748 TI - Tongue reconstruction with a combined brachioradialis-radial forearm flap. AB - Total glossectomy adversely affects speech and swallowing, and subsequent reconstruction results in limited functional return. The radial forearm flap has been reliably used to resurface glossectomy defects, but has limited bulk with which to aid in palatoglossal contact for speech. The authors have modified the forearm flap by incorporating a segment of brachioradialis muscle, to increase bulk posteriorly and to aid in speech. Sufficient muscle perforators arise from the proximal brachial artery and enter the brachioradialis to permit transfer of the muscle with the fasciocutaneous forearm flap as a single free-flap unit. The muscle is folded onto itself and enclosed within the forearm flap skin to create a neotongue. Coaptation of the antebrachial cutaneous nerves can provide a senate flap. Successful transfer of the combined brachioradialis/forearm flap in a patient who had undergone total glossectomy resulted in a neotongue good shape. Speech was rated good by a speech pathologist, and palatoglossal contact was observed on cineoradiograph. No functional loss at the donor site occurred. Inclusion of the brachioradialis muscle with the radial forearm flap as a combined unit results in a neotongue with good form and increased bulk posteriorly at the base, compared to a standard fasciocutaneous flap alone. This is a useful variation of the forearm flap. Sensory return is possible if the medial and/or lateral antebrachial cutaneous nerves of the flap are coapted to the lingual nerve. PMID- 10668749 TI - Documentation of brachial plexus compression in the thoracic inlet with quantitative sensory testing. AB - This study evaluated the cutaneous pressure threshold of subjects with and without a clinical diagnosis of brachial plexus compression in the thoracic inlet, usually termed thoracic outlet syndrome (TOS). Sixty-one subjects (102 arms) made up the control population; 11 subjects 120 arms) made up the TOS population. Assessment by two upper-extremity specialists preceded the testing. The one-point cutaneous pressure threshold was measured with the Pressure Specifying Sensory Device (PSSD) on the pulp of both the index and little finger (upper and lower brachial plexus distributions) with the arm in the unprovoked (adducted) and provoked (abducted 180 degrees) positions. In the control subjects, there was no significant change in the cutaneous pressure threshold between unprovoked and provoked positions. In contrast in patients with TOS, there was a significant increase in the cutaneous pressure threshold at both sites (p < .0001 ) between the unprovoked and the provoked positions. Furthermore, the cutaneous pressure threshold for patients with TOS was significantly higher in both positions than it was in the controls (p < .0001 ). It was concluded that measurement of changes in the cutaneous pressure threshold with the PSSD in distal sensory targets of the upper and lower trunk can identify patients symptomatic for compression, when the brachial plexus is provoked as part of the testing sequence. PMID- 10668750 TI - Posterior radial collateral artery as the basis of the lateral forearm flap. AB - The lateral forearm flap is being increasingly used for covering minor-to moderate-sized defects for which soft and thin skin is required. Within the framework of an anatomic study carried out on 28 cadaveric arms, the authors investigated the principal artery that supplies blood to this flap namely, the posterior radial collateral artery (PRCA). They found that distal to the lateral epicondyle, the PRCA lies in a constant axial line lateral to the brachioradialis muscle. The average length of the artery distal to the epicondyle is 8 cm. Distal to the epicondyle side branches of the PRCA build an arterial plexus 6 cm (+/-3.5 cm) long and 5 cm (+/-1.1 cm) wide. The posterior cutaneous antebrachii nerve lies close to the artery. This permits the harvesting of a flap that is both innervated and has adequate vascular supply. PMID- 10668751 TI - Efficacy of the "baby-sitter" procedure after prolonged denervation. AB - This study was undertaken to evaluate whether 40 percent of the hypoglossal nerve, which showed optimal efficacy in restoring orbicularis oculi muscle (OOM) function after different percentages of partial neurectomy in a previous study would be effective after prolonged denervation time. Twenty Sprague-Dawley rats were divided into four groups. In first-stage surgery the left facial nerve of all animals was transected at the level of the stylomastoid foramen and main zygomatic branch. Group A (controls) consisted of animals with only left facial nerves transected (no repair). In Groups B, C, and D the facial nerve was transected and the facial musculature was denervated for a period of 4, 8, and 12 weeks respectively. During a second-stage procedure, a 40 percent neurectomy was performed on the hypoglossal nerve. Subsequently, a nerve transfer was performed by coaptations of a saphenous nerve graft to the neurectomized hypoglossal nerve and the main zygomatic branch of the facial nerve that innervated the OOM. Behavioral analysis of blink reflex, electrophysiology, and axon and motor end plate counts in Groups B, C, and D showed superior results compared to Group A. There was no statistically significant difference observed among Groups B, C, and D (p > 0.05). Despite the diminished number of axons in the zygomatic branch and motor end-plates in the orbicularis oculi muscle after 12 weeks of denervation, there was still sufficient muscle target recovery to effect some eye closure in all groups except the controls. This study demonstrated in this model that the 40 percent partial neurectomy of the XII to VII component of the "baby-sitter" procedure was effective even after prolonged denervation. PMID- 10668752 TI - Variations on the "baby-sitter" procedure for reconstruction of facial paralysis. AB - Four cases of fresh facial paralysis were treated using the "baby-sitter" procedure, with fairly satisfactory results obtained. This procedure involves a combination of cross-face nerve grafting and a nerve cross-over technique such as using a hypoglossal-facial nerve or accessory-facial nerve anastomosis. Two stages are required. Nerve cross-over and cross-face nerve grafting using the sural nerve are performed in the first stage. After 1 year nerve anastomosis between the stump of the cross-face nerve graft and that of a buccal branch in the paralyzed cheek is performed during the second stage. The recovery of facial animation through this method is good, with patients able to create a near natural smile. Electromyography demonstrates double innervation in the paralyzed cheek area. PMID- 10668754 TI - Biliary tract reconstruction using an autologous vein graft in rats. AB - Numerous biologic and synthetic materials have been used with limited success as an interposed graft to repair segmental common bile duct (CBD) defects. The authors report here that an autologous vein graft can be successfully used to correct a CBD deficit contingent on accurate microsurgical technique immediate stenting and rapid graft vascularization. Thirty Sprague-Dawley rats underwent laparotomy and the experimental group (n=25) had a 3-mm segment of the CBD excised. The CBD defect was repaired using an interposed femoral vein graft aided by a plastic stent. The control group (n=5) had the CBD cut and repaired by means of primary anastomosis. The experimental group was subdivided into three sub groups each examined at three different postoperative intervals: 1, 4 and 12 weeks. The results showed that inflammation was apparent in the venous wall following the first postoperative week. A progressive loss of the vascular endothelium and replacement with the columnar epithelium typical of the CBD was seen in the vein graft. Nineteen of the 25 experimental rats (76 percent) of the animals survived without complication from the surgery and there were no abnormalities in the liver biochemical tests of these animals. Any biliary tract obstruction that developed was attributed to dislocation of the stent leading to collapse of the vein graft (experimental group), or constriction of the anastomosis (control group). This study demonstrates that biliary tract reconstruction using an autologous vein graft can be successfully performed in a rat model of CBD repair. The application of this method to the clinical setting is also discussed. PMID- 10668753 TI - Accelerated flap prefabrication with vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is a potent promoter of angiogenesis that has been shown to enhance revascularization of ischemic tissues, including skin flaps. This study was designed to investigate the value of a single topical application of recombinant human VEGF to accelerate flap viability in a rat model of a non-ischemic prefabricated flap. Prefabricated flaps were created in 48 Sprague-Dawley rats. An autologous tail artery loop was anastomosed to the femoral artery and vein, and implanted subcutaneously in the lower abdomen. Flaps were divided into two groups of 24 each. At the time of loop implantation the control group received 0.9 percent NaCl or a 16 percent vol/wet polyvinyl alcohol (PVA) solution: the treatment group received VEGF in 0.9 percent NaCl or VEGF in PVA. The PVA gel was used to facilitate topical application In each group, 3- x 4 cm flaps nurtured by the tail artery pedicle were elevated and resutured into place after 3, 4, and 5 weeks. The percentage of surviving skin of each flap was determined by planimetry 7 days after flap elevation. Mean skin survival areas at 3, 4, and 5 weeks were control group 0 percent. 8 percent and 17.5 percent; and VEGIF-treated group, 6 percent, 40 percent, and 66.7 percent respectively VEGF significantly improved flap survival by 5 weeks (p = 0.02). These results suggest that VEGF can accelerate maturation of prefabricated flaps. This approach could expand the application of flap prefabrication as a resource for reconstructive surgery. PMID- 10668755 TI - Composite tissue allotransplantation: a comprehensive review of the literature- part 1. PMID- 10668756 TI - Mind, muscles and motoneurones. AB - This review considers some of the adaptations which take place in the central nervous system to allow optimal performance of the musculoskeletal system for the smallest to the largest "efforts". Mental imagery of exercise helps performance but the way in which it works is multifactional: it evokes muscle contraction sufficient to activate muscle receptors. Furthermore, it is possible for subjects to focus specifically on control of particular muscles even without feedback from them. On the other hand maximal voluntary efforts, at least in isometric and in concentric contractions, can drive the motoneurones sufficiently to ensure full force production by the muscle. Many neural factors contribute to maintain force output during repetitive activity, including a feedback loop whereby increased central command during fatigue acts to enhance muscle perfusion. As peripheral muscle fatigue develops, changes occur in the excitability of the motor cortex. Recent evidence suggests that "central" factors leading to reduced drive to muscles in isometric contractions act "upstream" of motor cortical output. PMID- 10668757 TI - Physiological determinants of endurance exercise performance. AB - Performance in endurance events is typically evaluated by the power or velocity that can be maintained for durations of 30 min. to four hours. The two main by products of intense and prolonged oxidative metabolism that can limit performance are the accumulation of hydrogen ion (i.e. lactic acidosis) and heat (i.e. hyperthermia). A model for endurance performance is presented that revolves around identification of the lactate threshold velocity which is presented as a function of numerous morphological components as well as gross mechanical efficiency. When cycling at 80 RPM, gross mechanical efficiency is positively related to Type I muscle fiber composition, which has great potential to improve endurance performance. Endurance performance can also be influenced by altering the availability of oxygen and blood glucose during exercise. The latter need forms the basis for ingesting carbohydrate at 30-60 grams per hour during exercise. In laboratory simulations of performance, athletes fatigue due to hyperthermia when esophageal is approximately 40 degrees C, in association with near maximal heart rate and perceived exertion. It is likely that the central nervous system is involved in the aetiology of fatigue from hyperthermia. Dehydration during exercise promotes hyperthermia by reducing skin blood flow, sweating rate and thus heat dissipation. The combination of dehydration and hyperthermia during exercise causes large reductions in cardiac output and blood flow to the exercising musculature, and thus has a large potential to impair endurance performance. Endurance performance is optimized when training is aimed specifically at developing individual components of the model presented and nutritional supplementation prevents hypoglycemia and attenuates dehydration and hyperthermia. Indeed, the challenge at the transition to a new millennium is to synergistically integrate these physiological factors in training and competition. PMID- 10668758 TI - Optimisation of the biology of soft tissue repair. AB - As identified in this review, over the past twenty years there have been a number of very exciting new developments in the quest to optimise soft tissue repair. Comparing fetal soft tissue injuries, which heal by regeneration, to the adult processes of healing by inflammation-induced scar formation has led to a number of insights into how the latter may be improved. Seeding wounds with embryonic stem cells, bridging gaps with cell-derived "engineered tissues", addition of exogenous hyaluronic acid and modification of wounds to either enhance the growth factors which have been implicated in regeneration (e.g. TGF-B3) or block those implicated in scar formation (eg. TGF-B1) have all shown promise. Our group has quantified numerous cellular, molecular, biomechanical and matrix abnormalities of scar in a rabbit model of ligament healing. Based on these studies which we review here, three matrix deficiencies have been identified which appear to have specific implications to scar weakness: organisational "flaws", abnormal hydroxypyridinoline collagen cross-link densities and abnormally small, slow maturing collagen fibrils. In tests aimed at finding therapeutic solutions in this model, the addition of a 7ug bolus of TGF-B1 at day 21 or 2.5ng/day of TGF B1 being pumped into a wound x 21 days increased the size of ligament scars but did not improve their material strength. It also did not alter any of the above noted matrix deficiencies. A liposome-mediated anti-sense gene therapy approach aimed at decreasing the expression of the proteoglycan decorin in 21-day scars, however, has significantly increased the size of scar collagen fibrils as well as improved these scars mechanically. Based on these positive results from a single dose of only one targeted molecule, we believe that this gene therapy approach has great potential for further scar improvement. If combined with some of the other biological strategies reviewed above, a repair which is closer to true regenerative healing of ligaments, and all soft tissues, may eventually be achieved. PMID- 10668759 TI - Biological reaction to vibration--implications for sport. AB - In many situations of everyday life, vibration load occurs. Here whole body vibration in vehicles, such as boats, cars, helicopters and others as well as hand-transmitted vibration (motor saws etc.) can be named. As vibration is assumed liable to cause various threats to human health, a great number of studies in work science focussed on dose-effect relations and concepts for prevention. Although in many sports remarkable vibration load also occurs, there is very little research on the potential dangers and benefits of vibration stimuli, e.g. on whole body vibration and the implications for muscular activity and neuromuscular control in sport. In personal studies the damping behaviour and training effects under whole body vibration were investigated. Various research areas have been studied in order to approach the relevant topics: neuromuscular and posture control, energy metabolism in terms of oxygen uptake under whole body vibration and local concentration of phosphates by means of 31P-MRS. Furthermore the effects of a strength training under whole body vibration were analysed. The results underline that vibration is a neglected research topic in sport science from the preventive point of view as well as from the one focussing on the improvement of sport performance. PMID- 10668760 TI - Stress, anxiety and performance. AB - Research which has examined the catastrophe models of anxiety and performance is discussed. The conclusion drawn is that the evidence supports the notion of hysteresis, and partially supports the interactive effects prediction of the catastrophe models. Two potential explanations of anxiety induced performance catastrophes are then examined, processing efficiency theory and the conscious processing hypothesis. Process efficiency theory proposes that, as well as reducing attentional resources, cognitive anxiety can also lead performers to invest additional effort in the task in an attempt to allay their concerns and fears. Thus, performance may be maintained (or even enhanced), but at an increased physiological cost. Evidence is presented in support of the prediction that cognitive anxiety leads to increased effort. The conscious processing hypothesis proposes that anxious performers regress to an earlier stage of learning when they controlled movements using conscious processes and explicit knowledge. Evidence is presented from a number of recent studies which at least partially support this notion. Finally, it is suggested that these two different explanations are not necessarily mutually exclusive. PMID- 10668761 TI - Exercise and the immune system--influence of nutrition and ageing. AB - In essence, the immune system is enhanced during moderate and severe exercise, and only intense long-duration exercise is followed by impairment of the immune system. The latter includes suppressed concentration of lymphocytes, suppressed natural killer cell activity, lymphocyte proliferation and secretory IgA in saliva. During the time of immune impairment, referred to as "the open window", microbial agents, especially viruses may invade the host and infections may be established. One reason for the "overtraining effect" seen in elite athletes could be that this window of opportunism for pathogens is longer and the degree of immunosuppression more pronounced. Alterations in metabolism and metabolic factors may contribute to exercise-associated changes in immune function. Reductions in plasma-glutamine concentrations, altered plasma-glucose level, free oxygen radicals and prostaglandins (PG) released by the elevated number of neutrophils and monocytes may influence the function of lymphocytes and contribute to the impaired function of the later cells. Thus, nutritional supplementation with glutamine, carbohydrate, anti-oxidants or PG-inhibitors may, in principle, influence exercise-associated immune function. Although several intervention studies have been performed, it is premature to make recommendations regarding nutritional supplementation to avoid post-exercise impairment of the immune system. PMID- 10668762 TI - Mechanisms of muscle injury after eccentric contraction. AB - Eccentric contractions of skeletal muscles produce injury and, ultimately, muscle strengthening. Current data suggests that the earliest events associated with injury are mechanical in nature and may be based primarily on the sarcomere strain experienced by the muscle. In this review, recent experimental data, primarily from rabbit dorsiflexor muscles, are used to provide general information regarding the factors that cause injury and means for preventing injury. Mechanical experiments reveal that excessive sarcomere strain is the primary cause of injury. We hypothesize that excessive strain permits extracellular or intracellular membrane disruption that may permit hydrolysis of structural proteins leading to the myofibrillar disruption that is commonly observed. Inflammation that occurs after injury actually further degrades the tissue, but prevention of the inflammation leads to a long-term loss in muscle function. Simple treatments such as increasing muscle oxidative capacity ("getting into shape") or cyclic stress-relaxation of tissue ("stretching out") have no measurable effect on the magnitude of muscle injury that occurs. Ultimately, an improved understanding of the damage mechanism may improve our ability to provide rehabilitative and strengthening prescriptions that have a rational scientific basis. PMID- 10668763 TI - Determination of optimal pacing strategy in track cycling with an energy flow model. AB - The purpose of this study was to investigate the effect of pacing strategies on performance times in the 1000 m time trial event and the 4000 m pursuit event in track cycling. For this purpose, we simulated these events with a model based on the flow of energy in cycling. Different strategies in distributing the available anaerobic energy were evaluated and we compared model predictions of split times and final times with values achieved by cyclists during championships. The best result at the 1000 m time trial was obtained when the cyclist had the highest anaerobic peak power output and used an 'all-out' strategy. The fastest time on the 4000 m pursuit was achieved with an 'all-out' start at a high level of initial power output, followed by a constant anaerobic power output after 12 seconds, resulting in an evenly paced race. The results show that even small variations in pacing strategy may have substantial effects on performance. There seems to be an opportunity to gain a competitive advantage when individual athletes experiment with small variations in pacing strategy to find the precise individual strategy that works best under specific conditions. PMID- 10668764 TI - The John Stanley Coulter lecture. Overcoming the odds: the health of women with physical disabilities in the United States. PMID- 10668765 TI - Functional electrical stimulation effect on orthostatic hypotension after spinal cord injury. AB - OBJECTIVE: To investigate the possibility of using functional electrical stimulation (FES) to control orthostatic hypotension in patients with spinal cord injury (SCI) and to clarify the mechanism of the response. DESIGN: Subjects were tilted by 10 degree increments with varying intensities of lower-extremity FES. Stimulation over muscles was compared to stimulation over noncontractile sites. SETTING: Physical therapy department of a major rehabilitation center. PATIENTS: Six patients with SCI above T6 (3 with recent injury recruited consecutively from an inpatient spinal cord rehabilitation unit, and 3 from the community with longstanding injury, recruited as volunteers). MAIN OUTCOME MEASURES: Blood pressure, heart rate, and perceived presyncope score recorded at each tilt angle and analyzed using a multivariate analysis of variance statistical methodology. RESULTS: Systolic and diastolic blood pressure increased with increasing stimulation intensities (systolic, p = .001; diastolic, p = .0019) and decreased with increasing angle of tilt (p < .001) regardless of the site of stimulation. Subjects tolerated higher angles of incline with electrical stimulation than without (p = .03). CONCLUSIONS: FES causes a dose-dependent increase in blood pressure independent of stimulation site that may be useful in treating orthostatic hypotension. PMID- 10668766 TI - Spasticity in spinal cord injury: self- and clinically rated intrinsic fluctuations and intervention-induced changes. AB - OBJECTIVES: (1) To determine patterns of intrinsic fluctuations in spasticity, using repeated self-ratings, in subjects with spinal cord injury (SCI); and (2) To determine the relation between self-ratings of spasticity using a visual analogue scale (VAS) and clinical ratings of spasticity using the Modified Ashworth Scale (MAS) before and after spasticity-reducing treatment. DESIGN: Part I: observational, prospective cross-sectional study; part II: experimental, prospective longitudinal study. SETTING: Outpatient clinic of the Karolinska Hospital, Stockholm, Sweden. PATIENTS: Forty-five persons with SCI (39 men, 6 women); mean age at injury, 26 yrs; mean time since injury, 11 yrs. INTERVENTION: Repetitive passive movements of standardized range of motion in three different body positions, performed by two motorized tables. MAIN OUTCOME MEASURES: VAS ratings of spasticity, every other hour when awake, and of movement-provoked spasticity, rated before and after each treatment session, and MAS ratings of movement-provoked spasticity, before and after each treatment session. RESULTS: The spasticity of cervical SCI subjects fluctuated significantly (p < .05) during the day, unlike the spasticity of thoracic SCI subjects. Immediately after intervention with passive movements, spasticity ratings in thoracic motor complete SCI patients decreased by 11 to 14 mm (90%, p < .001) as self-rated on VAS and by 1 to 2 grades (50%, p < .001) as measured with MAS. A 30% (p < .018) decrease in VAS values of intrinsic pattern of spasticity was maintained over time when treatment was given regularly and was maintained for at least 1 week after discontinuation of treatment. VAS ratings correlated significantly with MAS ratings (r = .44 to .62, p < .001). CONCLUSION: Repetitive passive movement intervention decreased spasticity when performed regularly, as assessed by VAS and MAS ratings. VAS and MAS ratings were significantly correlated. It is recommended that SCI patients repeatedly rate their spasticity to establish a baseline before and to track changes after interventions aimed at reducing spasticity. The time of day when spasticity is measured seems more important in cervically injured individuals, because of their more pronounced intrinsic fluctuations. PMID- 10668767 TI - Estimation of geometric properties of cortical bone in spinal cord injury. AB - OBJECTIVE: To evaluate structural and geometrical properties of the tibia shaft in subjects with spinal cord injury (SCI) and subjects without SCI and to estimate the potential usefulness of a multimodal approach to diagnosing osteoporosis in SCI. DESIGN: A cross-sectional study of randomly selected SCI and non-SCI subjects. METHODS: Measurements of bone geometric indices by computed tomography, and calculated bending stiffness with a biomechanical testing method. SETTING: An SCI center hospital. SUBJECTS: Ten men without known orthopedic or neurologic impairments (controls), 10 men with SCI who had a history of lower extremity pathologic fracture since SCI, and 10 men with SCI who had never had lower extremity pathologic fracture. RESULTS: Analysis of geometric and structural indices of subjects' tibias found a significant difference in all geometric indices between controls and the SCI subjects with pathologic fracture history. Between the controls and the SCI subjects with no fracture history, however, differences were found only in cross-sectional area and calculated bending stiffness. CONCLUSION: Structural analysis of leg bone, combined with measurement of bone density, may improve the ability to assess fracture risk in patients with SCI. PMID- 10668768 TI - Myosin heavy chain isoform and ubiquitin protease mRNA expression after passive leg cycling in persons with spinal cord injury. AB - OBJECTIVE: To determine the effects of passive leg cycling exercise on myosin heavy chain (MHC) isoform and ubiquitin (UBI) protease mRNA expression in patients with spinal cord injury (SCI). STUDY DESIGN: Case series. INTERVENTION: Eight SCI subjects (5 men, 3 women) participated in a 12-week exercise program involving the Psycle ergometer. Training occurred 2 days a week at 75% of each subject's maximum heart rate. Anthropometric measures (body weight, thigh girth, and body mass index) and muscle biopsy specimens were obtained before and after training. Analyses were performed to determine the mRNA expression of types I, IIa, and IIx MHC, as well as UBI, a UBI-conjugating enzyme (E2), and 20S proteasome (20S). RESULTS: Despite small increases, paired t tests (p < .05) to assess changes from pretraining to posttraining failed to locate significant differences for the three anthropometric measures. For mRNA expression, there were significant increases in expression of MHC types IIa and IIx and significant decreases in expression for UBI, E2, and 20S. CONCLUSION: Exercise using passive leg cycling increases the expression of fast MHC isoforms while concomitantly decreasing proteolytic activity associated with muscle degradation, thus helping to possibly ameliorate muscle atrophy in patients with SCI. PMID- 10668769 TI - Gabapentin effect on spasticity in multiple sclerosis: a placebo-controlled, randomized trial. AB - OBJECTIVE: To investigate the effect of gabapentin on subject self-report and physician-administered spasticity scales in individuals with multiple sclerosis. DESIGN: Prospective, double-masked, placebo-controlled, crossover design. SETTING: The Multiple Sclerosis Center at the Denver Veterans Affairs Medical Center. INTERVENTION: Subjects were titrated to either 900 mg gabapentin orally three times a day or placebo over a 6-day period. Subjects underwent a 14-day washout and then were crossed over. No other changes were made to their medication regimen. MAIN OUTCOME MEASURES: The outcome measures were divided into two categories: subject self-report scales physician-administered scales. Subject self-report scales included the spasm frequency scale, spasm severity scale, interference with function scale, painful spasm scale, and global assessment scale. Physician-administered scales included the Modified Ashworth Scale, clonus scale, deep tendon reflexes, plantar stimulation response, and the Kurtzke Expanded Disability Status (EDSS) Scale. Digit Span and Digit Symbol subtests of the WAIS-R Intelligence Scale were administered to assess for possible impaired concentration. The Fatigue Impact Scale was administered to assess for changes in fatigue. The adjective generation technique was administered to assess for alterations in mood. RESULTS: A statistically significant reduction in the impairment of spasticity was found in the gabapentin-treated subjects compared with placebo as measured by the self-report scales of the spasm severity scale, interference with function scale, painful spasm scale, and global assessment scale and by the physician-administered scales of the Modified Ashworth and plantar stimulation response. No significant difference was noted in the Digit Span, Digit Symbol, adjective generation technique, and EDSS. CONCLUSION: Gabapentin reduces the impairment of spasticity, compared with placebo, without the side effects of worsening concentration and fatigue. PMID- 10668770 TI - Cervical nonorganic signs: a new clinical tool to assess abnormal illness behavior in neck pain patients: a pilot study. AB - OBJECTIVE: To develop and assess the reliability of a group of cervical nonorganic physical signs to be used as a simple screening tool for identifying patients with low neck pain who exhibit abnormal illness behavior. DESIGN: Survey, consecutive sample. DATA SET: Double masked. SETTING: Functional restoration program. PATIENTS: Twenty-six consecutive patients with complaints of chronic neck pain (greater than 4 months duration). Each patient was evaluated by a physician and then again by either a physical or occupational therapist, for the presence of specific cervical nonorganic signs. Both of the evaluations occurred on the same day. MAIN OUTCOME MEASURES: Five categories consisting of eight tests were appraised: (1) tenderness, (2) simulation, (3) range of motion, (4) regional disturbance, and (5) overreaction. RESULTS: The percent agreement between raters ranged from a high of 100% for regional sensory disturbance, to a low of 68% for one of the simulation tests. The average agreement between raters across all of the nonorganic test signs was 84.6%. Likewise, kappa coefficients ranged from 1.00 to .16, reflecting differences in strength of agreement. CONCLUSION: For many years, the lumbar nonorganic signs (developed by Waddell and colleagues) have been a useful screening tool in the assessment of abnormal illness behavior in the low back pain population. For the first time, a group of cervical nonorganic signs have been developed, standardized, and proven reliable. PMID- 10668771 TI - Peripheral plasma amino acid abnormalities in rehabilitation patients with severe brain injury. AB - OBJECTIVE: Acute severe brain injury causes an increased mobilization of amino acids from tissue. The plasma amino acid profile of patients undergoing rehabilitation after brain injury is unknown. This study was aimed at delineating the plasma amino acid profile of rehabilitation patients with brain injury. DESIGN: Peripheral plasma aminogram, lactate, pyruvate, glycerol, ketone body, and carnitine concentrations were determined in 11 patients with brain injury (34.6+/-15 years old, 60+/-16.8 days after injury) and in 8 controls. Resting energy expenditure and nitrogen balance were also determined. RESULTS: (1) All essential amino acids and about 50% of nonessential amino acids were significantly lower in brain injury patients than in controls (p < .05). (2) Plasma amino acids were lower irrespective of either energy and protein intake or nitrogen balance. (3) Total carnitine concentration and esterified/free carnitine ratio were higher in brain injury patients than in controls (p < .05). CONCLUSIONS: Rehabilitation patients with brain injury may have an important reduction of their plasma aminogram. Muscle tissue depletion and the persistence of a hypercatabolic state caused by subclinical infections, pressure sores, and immobility may contribute to this reduction. PMID- 10668772 TI - Barriers to exercise in African American women with physical disabilities. AB - OBJECTIVE: To examine what factors African American women with one or more physical disabilities perceive as barriers to exercise and how they rank them. SETTING: Department of Disability and Human Development at a major university. STUDY DESIGN: Data were collected through telephone interview using a newly developed instrument (Barriers to Physical Exercise and Disability [B-PED]) that addressed issues related to physical activity and the subjects' disability. SUBJECTS: Fifty subjects were asked questions about their participation and interest in structured exercise. RESULTS: The four major barriers were cost of the exercise program (84.2%), lack of energy (65.8%), transportation (60.5%), and not knowing where to exercise (57.9%). Barriers commonly reported in nondisabled persons (eg, lack of time, boredom, too lazy) were not observed in our sample. Only 11% of the subjects reported that they were not interested in starting an exercise program. The majority of subjects (81.5%) wanted to join an exercise program but were restricted by the barriers reported. CONCLUSION: African American women with a physical disability are interested in becoming more active but are limited in doing so because of their inability to overcome several barriers to increased physical activity participation. PMID- 10668773 TI - Assessing joint pain complaints and locomotor disability in the Rotterdam study: effect of population selection and assessment mode. AB - OBJECTIVE: To assess the prevalence of self-assessed and physician-assessed disability and joint pain, their association, and the effect of cohort reduction and mode of assessment. DESIGN: Cross-sectional population survey. SETTING: General population, age 55 years and older. SUBJECTS: Independently living participants of the Rotterdam Study, including 1,156 men and 1,739 women. OUTCOME MEASURES: Self-reported and physician-assessed joint complaints. Patients' self assessment of locomotor disability was by response to questions from the Stanford Health Assessment Questionnaire; physicians assessed patients' disability by administering activity tests. RESULTS: Reduction of the study cohort because of nonresponse and missing data had no influence on the frequency and effect measures. The physician-assessed prevalence of pain of the hips, knees, or feet was significantly lower than the self-assessed prevalence, with the percentage agreement being 83% for men and 74% for women, with kappa-values of approximately .40. The prevalence of physician-assessed locomotor disability was also significantly lower than the self-assessed disability, with the percentage agreement being 83% for men and 78% for women, with kappa values of .41 and .47, respectively. The associations of joint complaints with disability were similar for both modes of assessment. CONCLUSION: Cohort reduction caused by nonresponse and missing data had no influence on estimates of frequency and association. Self assessment gives higher prevalences of joint complaints and locomotor disability than physician assessment, but the associations between complaints and disability were the same. PMID- 10668774 TI - Asymmetry of gait initiation in patients with unilateral knee arthritis. AB - OBJECTIVE: To identify how patients with knee arthritis modify their equilibrium and movement control strategies during gait initiation. DESIGN: Observational study. SETTING: University hospital movement analysis laboratory. PARTICIPANTS: Twelve patients with unilateral knee arthritis and 12 healthy control subjects. MAIN OUTCOME MEASURES: Durations of the phases of gait initiation (ie, postural, monopodal, and double-support phases), center-of-pressure displacements, ground reaction forces, pelvic velocity, step length, and knee range of motion were measured using a movement analysis system and force plates. RESULTS: Gait initiation was slower in patients than in controls no matter which leg was the supporting one. In patients, the durations of the postural and the monopodal phases were modified in an asymmetrical way according to the leg used as the supporting one. The postural phase was lengthened and the monopodal phase was shortened when the affected leg was the supporting one. Opposite effects were observed when the sound leg was supporting. Step length, knee range of motion, and maximal pelvic velocity were reduced in patients whatever the side of the supporting leg. CONCLUSION: Gait initiation is an asymmetrical process in unilateral knee arthritis patients, who develop adaptive posturomotor strategies that shorten the monopodal phase on the affected leg. PMID- 10668775 TI - Measurement of brake response time after right anterior cruciate ligament reconstruction. AB - OBJECTIVE: Recommendations on safe driving after anterior cruciate ligament (ACL) reconstruction have been largely intuitive. We studied brake response time in patients who participated in outpatient rehabilitation after right ACL reconstruction. DESIGN: Prospective, repeated measures design comparing 14 patients post-ACL reconstruction with 21 subjects with normal knees. INTERVENTIONS AND MAIN OUTCOME MEASURES: The following measures were assessed every 2 weeks for 10 weeks: brake response time, 6-meter walk time, knee range of motion, pain, and joint effusion. Statistical testing used analysis of covariance with repeated measures. Significant variables were analyzed separately and post hoc tests conducted using the least squares differences method. Both groups were compared with published norms from the American Automobile Association. RESULTS: No significant gender differences across main effects were detected. Brake response times for men improved significantly after week 6 (p < .05) and week 10 (p < .01). Brake response times for women in the ACL treatment group matched controls at 6 weeks. Six-meter walk times for control subjects were faster than those for the ACL group preoperatively (2.6 sec vs 5.5 sec), but equalized by week 6. CONCLUSIONS: After right ACL reconstruction, brake reaction times of rehabilitated men and women matched those of established controls after 4 to 6 weeks. Measuring brake response times during rehabilitation may ensure that individual patients return to driving in a safe and timely manner. PMID- 10668776 TI - Social issues in the rehabilitation of younger stroke patients. AB - OBJECTIVES: To study social factors and outcomes in stroke rehabilitation patients under the age of 50. STUDY DESIGN: Retrospective chart review examining (1) martial status and employment status on admission and at 3 months post discharge, (2) discharge destination, (3) the presence of absence of children under the age of 16, and (4) psychosocial difficulties as recorded by staff during hospitalization. SUBJECTS AND SETTING: Eighty-three consecutive stroke patients under the age of 50 admitted to a Canadian tertiary-care hospital rehabilitation unit. MAIN OUTCOME MEASURES: Discharge destination and primary caregiver at discharge, and return to work and marital separation 3 months after rehabilitation discharge. RESULTS: Of the 55 patients with spouses, 8 (14.5%) separated within 3 months of hospital discharge. Fifteen of the 83 patients (18.1%) were not able to return to their premorbid place of residence; 4 (4.8%) required institutionalization. Of the 64 patients employed outside the home or studying at the time of their stroke, only 13 (20.3%) were able to return to work within 3 months of their discharge to home. Only 9.4% of those working full-time were able to return to full-time employment. CONCLUSIONS: Rehabilitation of young stroke patients is associated with a variety of social problems, including marital breakup, child care responsibilities, and return to employment, which are uniquely important in this age group. PMID- 10668777 TI - Joint position during anterior-posterior glide mobilization: its effect on glenohumeral abduction range of motion. AB - OBJECTIVE: To investigate the effect of joint position during an anterior posterior glide (APG) procedure on the range of motion (ROM) of glenohumeral abduction in cadaver specimens. DESIGN: Mechanical simulation of APG mobilization and abduction torque ROM measurement of the glenohumeral joint with a material testing system. The immediate mechanical efficacy of APG was compared in two groups of specimens at two different joint positions: midrange (n = 5) and end range (n = 6) of glenohumeral abduction. SETTING: Biomechanics laboratory. SPECIMENS: Eleven fresh cadaver shoulder specimens (mean age, 66.9+/-2.5 yrs). MAIN OUTCOME MEASURE: Improvement in glenohumeral abduction torque ROM obtained before and after APG procedure. RESULTS: Glenohumeral abduction improved significantly, as indicated by a significant increase (Kruskal-Wallis statistics, chi2 = 7.50, p = .006) in the torque ROM of the end range group (mean +/- standard error of the mean, 2.02 degrees +/- .20 degrees) over the midrange group (.64 degrees +/- .08 degrees). A significant difference in the magnitude of peak displacement of the humeral head between midrange (14.44+/-3.56 mm) and end range (3.19+/-.81 mm) groups was also found (Mann-Whitney test, p < .030). CONCLUSION: This study demonstrated that APG technique performed at close to the end of the range of abduction is more effective in improving glenohumeral abduction ROM than that performed at the middle of the range of abduction. PMID- 10668778 TI - Clinical tests of standing balance: performance of persons with multiple sclerosis. AB - OBJECTIVE: To investigate differences in performance between people with multiple sclerosis (MS) and control subjects on clinical tests of balance, and to assess performance consistency on balance tests in people with MS from morning to afternoon. STUDY DESIGN: Two factor repeated measures design with a two group sample of convenience. SETTING: Kingston Centre and the Camberwell Centre of the MS Society of Victoria, Australia. SUBJECTS: Fourteen people with MS and 14 control subjects matched for age, height, and sex. MAIN OUTCOME MEASURES: Subjects were measured on their ability to maintain standing balance in steady stance, (feet apart, feet together, stride stance, tandem stance, and single leg stance), during self-generated perturbations (functional reach, arm raise, and step tests) and in response to an external perturbation. Participants with MS were also asked to rate their fatigue level in the morning and afternoon. RESULTS: There were no differences between MS and control groups on the ability to maintain standing balance with feet apart, feet together, or in stride stance. Participants with MS performed more poorly than control subjects in tandem stance and single leg stance and in the functional reach test, arm raise test, step test, and in response to an external perturbation. There was little change in balance from morning to afternoon in participants with MS (ICCs (2,1) .70 to .94), despite an increase in self-rated fatigue (t(14) = -3.14, p = .008). CONCLUSION: The ability to maintain balance in standing is a marked problem in people with MS despite the consistency of their performance from morning to afternoon. PMID- 10668779 TI - Catecholamine-induced hypertension in lumbosacral paraplegia: five case reports. AB - Hypertension in the patient with SCI is relatively rare and generally restricted to patients with high-level injuries where autonomic dysreflexia can occur. Resting blood pressure in individuals with SCI has been described as lower than that in the normal population. This report describes five previously normotensive teenagers with subsequent paraplegia as a result of gunshot wounds who presented with hypertension secondary to idiopathic elevation of plasma or urinary catecholamine levels. A clonidine suppression test was used as a neuroprobe to inhibit centrally mediated sympathetic outflow, excluding the probability of an extra-axial autonomous catecholamine-secreting tumor as the possible source of hypertension. Positive suppression was achieved in four patients (41%, 37.2%, 4.8%, and 37.2% decreases). One patient had values corresponding to orthostatic changes (an increase of 63%) because of poor compliance with the test. This patient was lost to follow-up; in the remaining four, hypertension resolved at 12, 8, 9, and 6 weeks postinjury. The increased circulating catecholamine level appears to be promoted by a centrally mediated response to the SCI. Elevated blood pressure probably results from an upgraded receptor regulation or an increased receptor sensitivity on the affected cells in the absence of restraining spinal reflexes. The pathophysiology of such hypertension seems to be secondary to autonomic dysfunction and, although it may be transient, it should be treated promptly and reevaluated periodically until stabilization is achieved. PMID- 10668780 TI - Aerobic training in a patient with nonsevere aplastic anemia: a case report. AB - This case report examined whether a 26-year-old man with a 5-year history of nonsevere aplastic anemia could perform aerobic training and whether exercise was beneficial. Testing was performed at baseline and at 8 and 16 weeks and included complete blood tests, graded exercise tests with breath-by-breath gas analyses, and health status assessment with the Medical Outcomes Survey SF-12 health survey. Training consisted of treadmill walking for 25 minutes, 3 days a week for 16 weeks, at 75% of maximal heart rate. The patient successfully completed 16 weeks of training and had no adverse effects from testing or training. Training did not produce changes in disease-related measures (hematologic values) or impairment measures (cardiopulmonary measures of fitness). The mental component of the SF-12 improved from below 2 standard deviations from the population mean to within 1 standard deviation of the population mean. The benefits of aerobic training for this person with aplastic anemia were that he showed that he could participate in aerobic-type activities and that training appeared to improve his mental health. PMID- 10668781 TI - Cardiopulmonary rehabilitation in a patient with Noonan syndrome. AB - Noonan syndrome, an autosomal dominant disease occurring with an incidence of 1 in 1,000 to 1 in 2,500 live births, is characterized by its particular cardiovascular abnormalities, including pulmonic valve stenosis, pulmonary artery stenosis, and, more rarely, septal defects and coarctation of the aorta. The case of a 20-year-old man admitted for inpatient cardiopulmonary rehabilitation after pulmonic valve repair, left pulmonary artery angioplasty, and pectus excavatum repair is presented. His endurance was markedly decreased, thus limiting his ability to perform activities of daily living and reducing his exercise tolerance. With participation in a comprehensive cardiopulmonary rehabilitation program, he experienced marked improvement with independence in his activities of daily living and an increase in his metabolic equivalent levels from to 2.8 to 5.4. After inpatient rehabilitation, he underwent left pulmonary stent placement before being discharged home. Subsequent outpatient cardiopulmonary rehabilitation has continued to improve significantly his overall exercise tolerance. Given that Noonan syndrome is viewed as the most common syndrome associated with congenital heart disease after Down syndrome, physiatrists must be familiar with its presentation, its associated abnormalities, and the treatment approach to optimize the patient's cardiopulmonary, musculoskeletal, and psychological status. PMID- 10668782 TI - The rehabilitation marketplace: economics, values, and proposals for reform. AB - This article examines whether the ideals, goals, and values of medical rehabilitation can be realized in a market-based health care system. The article observes that rehabilitation is greatly disadvantaged in today's health care marketplace, which violates virtually all the assumptions of a perfectly competitive one. Nevertheless, the authors argue that rehabilitation goals and market economics are not inherently incompatible and call for several market reforms that are congruent with both rehabilitation goals and market theory. These reforms will clarify and facilitate providers' fiduciary responsibilities to patients as well as their accountability to payers. The authors conclude that while the marketplace is an inevitable medium for realizing rehabilitation goals, the vision and value of rehabilitation will not derive from the internal workings of the marketplace but ultimately from committed individuals and socially responsible institutions outside the marketplace. PMID- 10668783 TI - Muscle strength and vitamin D. PMID- 10668784 TI - Wheelchair spotter straps. PMID- 10668785 TI - Duplex ultrasound screening. PMID- 10668786 TI - Upregulation of L-plastin gene by testosterone in breast and prostate cancer cells: identification of three cooperative androgen receptor-binding sequences. AB - L-Plastin is normally a leukocyte-specific actin-binding protein; it is also expressed in the majority of human cancer cell lines that are derived from many types of solid tumors. We have previously reported the isolation of the L-plastin gene promoter, in which we identified several potential steroid receptor-binding sequences. We now obtained evidence that L-plastin gene expression was positively regulated by testosterone in androgen receptor (AR)-positive prostate and breast cancer cells. DNase I footprint analysis identified three AR-binding elements (ARE) located in a 545-bp region approximately 1.1 kb upstream from the transcription initiation site. However, each of these three AREs exhibited very little testosterone/AR-responsive enhancer activities toward a test promoter (of the thymidine kinase gene) when tested in MCF-7 breast cancer cells. Their testosterone/AR responsiveness became evident only when two or three of them were combined. In PC-3 prostate cancer cells, cooperation among L-plastin AREs was still evident although individually they had moderate levels of testosterone/AR responsiveness. Thus, the three L-plastin AREs, despite their imperfect sequences compared with the consensus ARE, could cooperate with each other to become a potent testosterone/AR-responsive unit, which was likely responsible for the inducibility of the L-plastin gene by testosterone. PMID- 10668787 TI - C/EBPalpha is required to maintain postmitotic growth arrest in adipocytes. AB - Terminal differentiation is often coupled with irreversible loss of proliferative potential. The CCAAT enhancer binding protein alpha (C/EBPalpha) preferentially accumulates in postmitotic, differentiated 3T3-L1 adipocytes but declines during tumor necrosis factor alpha (TNFalpha)-induced dedifferentiation. We have discovered that this decline in C/EBPalpha correlates with an increased mitotic growth potential. In order to further investigate the antimitotic activity of C/EBPalpha, we introduced antisense C/EBPalpha RNA into 3T3-L1 cells to block endogenous C/EBPalpha expression. When treated according to the standard differentiation protocol, stable cells lines harboring antisense C/EBPalpha RNA did not differentiate into fat-laden adipocytes, consistent with previous findings (Lin F, Lane MD, Genes Dev 1992;6:533-544). We found that these undifferentiated cells expressing antisense-C/EBPalpha can reenter the cell cycle after mitogenic stimulation at a time in development when parental 3T3-L1 cells cannot. Moreover, the expression profiles of the growth-arrest-associated genes gas1 and gas2 revealed that the antisense C/EBPalpha-expressing cells withdrew from the cell cycle after the period of clonal expansion but failed to progress to the state of least proliferative potential characteristic of terminally differentiated adipocytes. PMID- 10668788 TI - Kappa opioid receptor endocytosis by dynorphin peptides. AB - Internalization and downregulation are important steps in the modulation of receptor function. Recent work with the beta2 adrenergic and opioid receptors have implicated these processes in receptor-mediated activation of mitogen activated protein kinase (MAPK). We have used CHO cells expressing epitope-tagged rat kappa opioid receptors (rKORs) and prodynorphin-derived peptides to characterize the agonist-mediated endocytosis of rKORs and activation of MAPK. Kappa receptor-selective peptides induced receptor internalization and downregulation whereas nonpeptide agonists did not. An examination of the ability of dynorphin A-17-related peptides (lacking C-terminal amino acids) to promote KOR internalization, inhibition of adenylyl cyclase, and MAPK phosphorylation revealed that the N-terminal seven residues play an important role in eliciting these responses. Both dynorphin peptides and nonpeptide agonists induced rapid and robust phosphorylation of MAPKs. Taken together, these results point to a difference in the ability of dynorphin peptides and nonpeptide ligands to promote rKOR endocytosis and support the view that rKOR internalization is not required for MAPK activation. PMID- 10668789 TI - Gamma-radiation-induced growth arrest and apoptosis in p53-null lymphoma cells is accompanied by modest transcriptional changes in many genes. AB - Damage to DNA produces cell cycle arrest, apoptosis, or both. The response in cells with p53 tumor suppressor function involves transcriptional changes, but whether that holds for cells lacking active p53, as in most tumors, is not known. Better characterization of the DNA damage response in tumors lacking p53 function is relevant to cytotoxic therapy. We have explored whether gamma-irradiated p53 null mouse T lymphoma cells undergo marked changes in transcription. Their arrest in G2/M prior to apoptosis required transcription. Transcripts whose abundance altered on irradiation were sought by subtractive hybridization, and 1010 candidate clones from two oppositely enriched cDNA populations were sequenced. Hybridization revealed small (<3-fold) increases or decreases in the transcripts of more than 15 genes, including some implicated in cell cycle control (e.g., BTG, Bap1) or apoptosis (e.g., STAT1, calpain), but no marked changes like those associated with other forms of T-cell death. Moreover, the expression of some critical apoptosis regulators, such as Bcl-2 family members, did not change. Hence, the G2/M arrest and apoptosis in the irradiated p53-null lymphoma appears to involve modest expression changes for many genes, but post-transcriptional alterations may be more critical. PMID- 10668790 TI - Downstream sequence adjacent to AUG affects translation of chloramphenicol acetyl transferase in eukaryotic cells. AB - The CAT gene is widely used as a reporter in eukaryotic systems because of the efficient translation of its mRNA. We report here that a sequence occurring in the CAT mRNA at +15 nucleotides from CAT AUG is essential for translation. This sequence includes a stem-loop structure, which, however, exhibits a calculated stability significantly lower than that required for a hairpin to act as an enhancer of translation in vitro. Replacement of this region with the corresponding sequence from mRNAs that are normally translated in eukaryotic systems drastically reduced translation of CAT in COS cells, although the consensus sequence around the AUG, known to be required for high-level translation initiation, was conserved. These observations may be relevant for the exploitation of the CAT reporter system for analysis of the mechanisms of translation initiation by means of fusion constructs. PMID- 10668791 TI - Molecular characterization of KMP11 from Trypanosoma cruzi: a cytoskeleton associated protein regulated at the translational level. AB - Kinetoplasmid membrane protein-11 (KMP11) is present in a wide range of trypanosomatids. In the present paper, we show that the T. cruzi KMP11 gene is organized in a cluster formed by four gene units arranged in a head-to-tail tandem manner located on a chromosome of about 1900 kb. Northern blot analyses indicated that the steady-state level of mature KMP11 transcripts of 0.52 kb is high and similar in the three forms of the parasite. The KMP11 mRNAs have a half life of about 16 h whose steady-state level is strongly downregulated when the parasites reach the stationary growth phase. The T. cruzi KMP11 sequence presents a significant homology with the amino-terminal third of the cytoskeleton associated protein CIP1 from Arabidopsis thaliana. Western blot and immunoelectron microscopy studies showed that KMP11 is present in the cytoskeleton structure. Because the strong downregulation observed in the de novo synthesis of KMP11 protein in parasites treated with vinblastine is not accompanied by a significant fall in the steady-state level of KMP11 mRNAs, regulatory control of the protein at the translational level is suggested. PMID- 10668792 TI - Atlantic salmon HNF-3/forkhead: cDNA sequence, evolution, expression, and functional analysis. AB - We report the isolation and characterization of a cDNA encoding an HNF-3 family member (as HNF-3) from Atlantic salmon (Salmo salar L). The important functional domains of HNF-3 proteins that have been characterized previously are revealed by segments of high identity along the alignment of the asHNF-3 with winged helix/forkhead amino acid sequences isolated from other species. A comparison of asHNF-3 cDNA and genomic DNA indicated that there were no introns present in the asHNF-3 gene. Expression of asHNF-3 protein in adult salmon tissues was not exclusive to liver but was also present in the pancreas and intestine. An RT-PCR analysis performed on salmon development showed that asHNF3 expression is detectable before gastrulation at the mid blastula transition stage. Functional analysis of the asHNF-3 protein using a characterized HNF-3 consensus binding site demonstrated that the protein can recognize and bind to specific HNF-3 consensus sequences. We also report the identification of a novel HNF3 binding site in the promoter of the Atlantic salmon transferrin gene. PMID- 10668793 TI - mRNA surveillance in mammalian cells: the relationship between introns and translation termination. PMID- 10668794 TI - On the wobble GoU and related pairs. AB - The wobble GoU pairs have been implicated in several biological processes where RNA molecules play a key role. We review the geometrical and conformational properties of wobble GoU pairs on the basis of available crystal structures of RNAs at high resolution. The similarities with the wobble A+oC pairs and UoU pairs are illustrated, while the differences with the recently discovered bifurcated G x U pairs are contrasted. PMID- 10668795 TI - Bimolecular exon ligation by the human spliceosome bypasses early 3' splice site AG recognition and requires NTP hydrolysis. AB - Here we report further characterization of an in vitro assay system for exon ligation by the human spliceosome in which the 3' splice site AG is supplied by a different RNA molecule than that containing the 5' splice and branch sites. By varying the time during splicing reactions when the 3' splice site AG is made available to the splicing machinery, we show that AG recognition need not occur until after lariat formation. Thus an early AG recognition event required for spliceosome formation and lariat formation on some mammalian introns is not required for exon ligation. Depletion/add-back studies and cold competitor challenge experiments reveal that commitment of a 3' splice site AG to exon ligation requires NTP hydrolysis. Because it both physically and kinetically uncouples exon ligation from spliceosome assembly and lariat formation, the bimolecular system will be a valuable tool for further mechanistic analysis of the second step of splicing. PMID- 10668796 TI - Organization of the 16S rRNA around its 5' terminus determined by photochemical crosslinking in the 30S ribosomal subunit. AB - The organization of the 5' terminus region in the 16S rRNA was investigated using a series of RNA constructs in which the 5' terminus was extended by 5 nt or was shortened to give RNA molecules that started at positions -5, +1, +5, +8, +14, or +21. The structural and functional effects of the 5' extension/truncations were determined after the RNAs were reconstituted. 30S subunits containing 16S rRNA with 5' termini at -5, +1, +5, +8 and +14 had similar structures (judged by UV induced crosslinking) and exhibited a gradual reduction in tRNA binding activity compared to that seen with 30S subunits reconstituted with native 16S rRNA. To create the 5' terminal site-specific photocrosslinking agent, the reagent azidophenacylbromide (APAB) was attached to the 5' terminus of 16S rRNA through a guanosine monophosphorothioate and the APA-16S rRNAs were reconstituted. Crosslinking carried out with the APA revealed sites in six regions around positions 300-340, 560, 900, 1080, the 16S rRNA decoding region, and at 1330. Differences in the pattern and efficiency of crosslinking for the different constructs allow distance estimates for the crosslinked sites from nucleotide G9. These measurements provide constraints for the arrangement of the RNA elements in the 30S subunit. Similar experiments carried out in the 70S ribosome resulted in a five- to tenfold lower frequency of crosslinking. This is most likely due to a repositioning of the 5' terminus upon subunit association. PMID- 10668797 TI - Specific HDV RNA-templated transcription by pol II in vitro. AB - RNA polymerase II is implicated in the RNA-templated RNA synthesis during replication of viroids and Hepatitis Delta Virus (HDV); however, neither the RNA template nor protein factor requirements for this process are well defined. We have developed an in vitro transcription system based on HeLa cell nuclear extract (NE), in which a segment of antigenomic RNA corresponding to the left hand tip region of the HDV rod-like structure serves as a template for efficient and highly specific RNA synthesis. Accumulation of the unique RNA product is highly sensitive to alpha-amanitin in HeLa NE and only partially sensitive to this drug in NE from PMG cells that contain an allele of the alpha-amanitin resistant subunit of pol II, strongly suggesting pol II involvement in this reaction. Detailed analysis of the RNA product revealed that it represents a chimeric molecule composed of a newly synthesized transcript covalently attached to the 5' half of the RNA template. Selection of the start site for transcription is remarkably specific and depends on the secondary structure of the RNA template, rather than on its primary sequence. Some features of this reaction resemble the RNA cleavage-extension process observed for pol II-arrested complexes in vitro. A possible involvement of the described reaction in HDV replication is discussed. PMID- 10668798 TI - A DNA target of 30 bp is sufficient for RNA-directed DNA methylation. AB - In higher plants, RNA-DNA interactions can trigger de novo methylation of genomic sequences via a process that is termed RNA-directed DNA methylation (RdDM). In potato spindle tuber viroid (PSTVd)-infected tobacco plants, this process can potentially lead to methylation of all C residues at symmetrical and nonsymmetrical sites within chromosomal inserts that consist of multimers of the 359-bp-long PSTVd cDNA. Using PSTVd cDNA subfragments, we found that genomic targets with as few as 30 nt of sequence complementarity to the viroid RNA are detected and methylated. Genomic sequencing analyses of genome-integrated 30- and 60-bp-long PSTVd subfragments demonstrated that de novo cytosine methylation is not limited to the canonical CpG, CpNpG sites. Sixty-base-pair-long PSTVd cDNA constructs appeared to be densely methylated in nearly all tobacco leaf cells. With the 30-bp-long PSTVd-specific construct, the proportion of cells displaying dense transgene methylation was significantly reduced, suggesting that a minimal target size of about 30 bp is necessary for RdDM. The methylation patterns observed for two different 60-bp constructs further suggested that the sequence identity of the target may influence the methylation mechanism. Finally, a link between viroid pathogenicity and PSTVd RNA-directed methylation of host sequences is proposed. PMID- 10668800 TI - Expanded CUG repeat RNAs form hairpins that activate the double-stranded RNA dependent protein kinase PKR. AB - Myotonic dystrophy is caused by an expanded CTG repeat in the 3' untranslated region of the DM protein kinase (DMPK) gene. The expanded repeat triggers the nuclear retention of mutant DMPK transcripts, but the resulting underexpression of DMPK probably does not fully account for the severe phenotype. One proposed disease mechanism is that nuclear accumulation of expanded CUG repeats may interfere with nuclear function. Here we show by thermal melting and nuclease digestion studies that CUG repeats form highly stable hairpins. Furthermore, CUG repeats bind to the dsRNA-binding domain of PKR, the dsRNA-activated protein kinase. The threshold for binding to PKR is approximately 15 CUG repeats, and the affinity increases with longer repeat lengths. Finally, CUG repeats that are pathologically expanded can activate PKR in vitro. These results raise the possibility that the disease mechanism could be, in part, a gain of function by mutant DMPK transcripts that involves sequestration or activation of dsRNA binding proteins. PMID- 10668799 TI - Interaction cloning and characterization of RoBPI, a novel protein binding to human Ro ribonucleoproteins. AB - Human Ro ribonucleoproteins (RNPs) are autoantigenic particles of unknown function(s) that consist of a 60-kDa protein (Ro60) associated with one hY RNA (hY1-5). Using a modified yeast three-hybrid system, named RNP interaction trap assay (RITA), we cloned a novel Ro RNP-binding protein (RoBPI), based on its property to interact in vivo in yeast with an RNP complex made of recombinant Ro60 (rRo60) protein and hY5 (rhY5) RNA. RoBPI cDNA contains three conserved RNA recognition motifs (RRM) and is present as a family of isoforms differing slightly at their 5' end. The 2.0-kb RoBPI mRNA was detected in all human tissues tested. Highly homologous cDNA sequences were found in banks of expressed sequence tags (ESTs) from mice. Two-hybrid, three-hybrid, and RITA experiments respectively established that 60 kDa RoBPI did not interact in yeast with rRo60 alone, with rhY5 RNA alone, or with bait RNPs consisting of rRo60 and recombinant hY1, hY3, or hY4 RNAs. RoBPI coimmunoprecipitated with Ro RNPs from HeLa cell extracts and partially colocalized with Ro60 in nuclei of cultured cells. Because hY5 RNA and RohY5 RNPs are recent evolutionary additions seen only in primates, but RoBPI seems more conserved, their interaction may represent a gain of function for Ro RNPs. Alternatively, interaction of RohY5 RNPs with RoBPI may have no functional bearing, but may underlie some of the unique biochemical and immunological properties of these RNPs. PMID- 10668801 TI - Fibrillarin binds directly and specifically to U16 box C/D snoRNA. AB - Eukaryotic nucleoli contain a large family of box C/D small nucleolar ribonucleoprotein complexes (snoRNPs) that are involved in processing and site specific methylation of pre-rRNA. Several proteins have been reported to be common factors of box C/D snoRNPs in lower and higher eukaryotes; nevertheless none of them has been clearly shown to directly interact with RNA. We previously identified in Xenopus laevis, by means of UV crosslinking in vivo, two proteins associated with box C/D snoRNAs, fibrillarin and p68. Here we show that fibrillarin interacts directly and specifically with the U16 box C/D snoRNA in a X. laevis oocyte nuclear extract and that it does not require p68 for binding. Specific binding is also obtained with a recombinant fibrillarin demonstrating that the protein is able to bind directly and specifically to U16 snoRNA by itself. PMID- 10668802 TI - Structural requirement for the two-step dimerization of human immunodeficiency virus type 1 genome. AB - Generation of RNA dimeric form of the human immunodeficiency virus type 1 (HIV-1) genome is crucial for viral replication. The dimerization initiation site (DIS) has been identified as a primary sequence that can form a stem-loop structure with a self-complementary sequence in the loop and a bulge in the stem. It has been reported that HIV-1 RNA fragments containing the DIS form two types of dimers, loose dimers and tight dimers. The loose dimers are spontaneously generated at the physiological temperature and converted into tight dimers by the addition of nucleocapsid protein NCp7. To know the biochemical process in this two-step dimerization reaction, we chemically synthesized a 39-mer RNA covering the entire DIS sequence and also a 23-mer RNA covering the self-complementary loop and its flanking stem within the DIS. Electrophoretic dimerization assays demonstrated that the 39-mer RNA reproduced the two-step dimerization process, whereas the 23-mer RNA immediately formed the tight dimer. Furthermore, deletion of the bulge from the 39-mer RNA prevented the NCp7-assisted tight-dimer formation. Therefore, the whole DIS sequence is necessary and sufficient for the two-step dimerization. Our data suggested that the bulge region regulates the stability of the stem and guides the DIS to the two-step dimerization process. PMID- 10668803 TI - In vivo misreading by tRNA overdose. AB - Rpb5-H147R is an AT-GC transition replacing CAC(His) by CGC(Arg) at a conserved and critical position of ABC27 (Rpb5p), one of the five common and essential subunits shared by all three eukaryotic RNA polymerases. This mutation is viable at 25 degrees C, but has a lethal phenotype at 34 degrees C. A search for dosage dependent suppressors identified five distinct clones that all bear a copy of the tRNA(His)GUG gene. Suppression was also observed with a small genomic insert bearing this tRNA gene and no other coding sequences, under conditions where there is a sevenfold increase in the cellular concentration of tRNA(His)GUG. Overexpressing tRNA(Arg)ICG, which normally decodes the suppressed CGC codon, counteracted suppression. Suppression is codon specific because it was abolished when replacing CGC by its synonymous codons CGA, CGU, or AGA, but was not detectably affected by several nucleotide substitutions modifying the surrounding sequence and is thus largely insensitive to the nucleotide context. It is proposed that overexpressing tRNA(His)GUG extends its decoding properties from CAC(His) to the noncognate CGC(Arg) codon through an illegitimate U x G pairing at the middle base of the anticodon. Accordingly, tRNA(His)GUG would compete with tRNA(Arg)ICG for chain elongation and generate a significant level of misreading errors under normal growth conditions. PMID- 10668804 TI - The SRm160/300 splicing coactivator subunits. AB - The SRm160/300 splicing coactivator, which consists of the serine/arginine (SR) related nuclear matrix protein of 160 kDa and a 300-kDa nuclear matrix antigen, functions in splicing by promoting critical interactions between splicing factors bound to pre-mRNA, including snRNPs and SR family proteins. In this article we report the isolation of a cDNA encoding the 300-kDa antigen and investigate the activity of it and SRm160 in splicing. Like SRm160, the 300-kDa antigen contains domains rich in alternating S and R residues but lacks an RNA recognition motif; the protein is accordingly named "SRm300." SRm300 also contains a novel and highly conserved N-terminal domain, several unique repeated motifs rich in S, R, and proline residues, and two very long polyserine tracts. Surprisingly, specific depletion of SRm300 does not prevent the splicing of pre-mRNAs shown previously to require SRm160/300. Addition of recombinant SRm160 alone to SRm160/300 depleted reactions specifically activates splicing. The results indicate that SRm160 may be the more critical component of the SRm160/300 coactivator in the splicing of SRm160/300-dependent pre-mRNAs. PMID- 10668805 TI - Mitochondrial minicircles in the free-living bodonid Bodo saltans contain two gRNA gene cassettes and are not found in large networks. AB - In trypanosomatids, the majority of the guide (g) RNAs that provide the information for U-insertion/deletion RNA editing are encoded by minicircles that are catenated into large networks. In contrast, in the distantly related cryptobiid Trypanoplasma borreli, gRNA genes appear to reside in large 180-kb noncatenated DNA circles. To shed light on the evolutionary history and function of the minicircle network, we have analyzed minicircle organization in the free living bodonid Bodo saltans, which is more closely related to trypanosomatids than T. borreli. We identified 1.4-kb circular DNAs as the B. saltans equivalent of minicircles via sequence analysis of 4 complete minicircles, 14 minicircle fragments, and 14 gRNAs. We show that each minicircle harbors two gRNA gene cassettes of opposite polarity residing in variable regions of about 200 nt in otherwise highly conserved molecules. In the conserved region, B. saltans minicircles contain a putative bent helix sequence and a degenerate dodecamer motif (CSB-3). Electron microscopy, sedimentation, and gel electrophoresis analyses showed no evidence for the existence of large minicircle networks in B. saltans, the large majority of the minicircles being present as circular and linear monomers (85-90%) with small amounts of catenated dimers and trimers. Our results provide the first example of a kinetoplastid species with noncatenated, gRNA gene-containing minicircles, which implies that the creation of minicircles and minicircle networks are separate evolutionary events. PMID- 10668807 TI - Strategies for the diagnosis of deep vein thrombosis and pulmonary embolism. PMID- 10668806 TI - The C-terminal domain of TAP interacts with the nuclear pore complex and promotes export of specific CTE-bearing RNA substrates. AB - Messenger RNAs are exported from the nucleus as large ribonucleoprotein complexes (mRNPs). To date, proteins implicated in this process include TAP/Mex67p and RAE1/Gle2p and are distinct from the nuclear transport receptors of the beta related, Ran-binding protein family. Mex67p is essential for mRNA export in yeast. Its vertebrate homolog TAP has been implicated in the export of cellular mRNAs and of simian type D viral RNAs bearing the constitutive transport element (CTE). Here we show that TAP is predominantly localized in the nucleoplasm and at both the nucleoplasmic and cytoplasmic faces of the nuclear pore complex (NPC). TAP interacts with multiple components of the NPC including the nucleoporins CAN, Nup98, Nup153, p62, and with three major NPC subcomplexes. The nucleoporin binding domain of TAP comprises residues 508-619. In HeLa cells, this domain is necessary and sufficient to target GFP-TAP fusions to the nuclear rim. Moreover, the isolated domain strongly competes multiple export pathways in vivo, probably by blocking binding sites on the NPC that are shared with other transport receptors. Microinjection experiments implicate this domain in the export of specific CTE-containing RNAs. Finally, we show that TAP interacts with transportin and with two proteins implicated in the export of cellular mRNAs: RAE1/hGle2 and E1B-AP5. The interaction of TAP with nucleoporins, its direct binding to the CTE RNA, and its association with two mRNP binding proteins suggest that TAP is an RNA export mediator that may bridge the interaction between specific RNP export substrates and the NPC. PMID- 10668808 TI - Prevention of venous thromboembolism. AB - Venous thromboembolism is the most common cause of preventable death among hospitalised patients. Systematic prophylaxis with antithrombotic agents in patients at risk for venous thromboembolism is the most effective approach to reduce morbidity and mortality. Despite this evidence, antithrombotic prophylaxis is still underused, due to the underestimation of incidence of venous thromboembolism and to the unjustified fear of bleeding complications. Both the characteristics of the individual patient and the clinical setting contribute to the definition of the risk for venous thromboembolism. Patient-related risk factors include clinical and molecular abnormalities. The grade of risk for venous thromboembolism is defined better by the clinical setting than by the patient characteristics. Prophylactic studies have been extensively carried out in surgical patients and, only more recently, in medical patients. Prophylactic methods include pharmacological agents, such as heparin, low molecular weight heparins, warfarin, and hirudin, as well as mechanical methods such as compression stockings and intermittent pneumatic compression. PMID- 10668809 TI - Treatment of venous thromboembolism. AB - The combination of heparin and oral anticoagulants has been the treatment of choice for most patients with venous thromboembolism in the last two decades. Heparin has been proven to be effective when administered by intravenous continuous infusion or by subcutaneous injection. Oral anticoagulants should be started at the same time and heparin should be discontinued when the levels of the International Normalized Ratio reach the therapeutic range, between 2.0 and 3.0. Low molecular weight heparin has potential advantages over heparin and can be administered in subcutaneous weight-adjusted fixed doses without need for monitoring. This has made home treatment of a large proportion of patients possible. Randomized clinical trials have demonstrated the efficacy and safety of this approach. The optimal duration of the secondary prophylaxis with oral anticoagulants is still a matter of debate. The rate of recurrence has been shown to be elevated, particularly in those patients with idiopathic venous thromboembolism. The presence of an active cancer or a thrombophilic state may require long-term anticoagulation, although not all the congenital hypercoagulable states seem to carry the same level of risk. A 3-month therapy is recommended when a transient risk factor is identified; lifelong treatment is recommended for patients with a second episode of venous thromboembolism. In all other cases, 6 months are currently recommended, but thereafter close monitoring of the patients is advisable. The use of different treatment strategies such as vena caval filter placement, thrombolysis, and surgical thrombectomy should be restricted to a limited number of situations. PMID- 10668810 TI - Companion animal medicine in the age of medical genetics. PMID- 10668811 TI - Mutation causing von Willebrand's disease in Scottish Terriers. AB - Von Willebrand's Disease (vWD) in the Scottish Terrier breed is a serious, often fatal, hereditary bleeding disorder. Elimination of the mutated gene by selective breeding is an important goal for the health of this breed. Although the standard protein-based tests are accurate for identification of affected Scottish Terriers, they are not reliable for the identification of carriers of the mutant gene unless multiple replicate assays are performed. A simple, highly accurate test for carriers of the disease is needed so that veterinarians can counsel clients on which animals to use in their breeding programs. The complete coding region of von Willebrand factor (vWF) complementary DNA (cDNA) was sequenced from an affected animal, and a single base deletion in the codon for amino acid 85 of the prepro-vWF cDNA that leads to Scottish Terrier vWD was identified. A highly accurate polymerase chain reaction assay was developed that can distinguish homozygous normal animals from those that are homozygous affected or heterozygous. In a voluntary survey of 87 animals provided by Scottish Terrier owners, 15 were carriers and 4 were affected with vWD, 2 of which had previously been shown to have undetectable vWF. The determination of the complete canine vWF cDNA sequence should facilitate the identification of additional vWD alleles in other breeds and other species. PMID- 10668812 TI - Metabolic and hormonal alterations in cats with hepatic lipidosis. AB - Hepatic lipidosis in cats is a commonly diagnosed hepatobiliary disease of unknown cause. The purpose of this prospective study was to characterize the blood hormone and lipid status of cats with hepatic lipidosis, and to compare this status to that of cats with other types of liver disease and to control cats. Twenty-three cats with hepatic disease were assigned to 1 of 2 groups on the basis of cytopathologic or histopathologic examination of the liver: group 1, hepatic lipidosis (n = 18); or group 2, cholangiohepatitis (n = 5). Ten healthy young adult cats were used as controls. Food was withheld from control animals for 24 hours before blood collection. Concentrations of plasma glucagon and serum insulin, cortisol, thyroxine, triglycerides, cholesterol, phospholipids, and nonesterified fatty acids (NEFAs) were determined in all cats, in addition to routine hematologic and serum biochemical testing. Cats with hepatic lipidosis had higher serum NEFA concentrations than cats with cholangiohepatitis or control cats (P < .05). Cats with cholangiohepatitis had higher serum cholesterol and phospholipid concentrations than those of cats with lipidosis or control cats (P < .05); their plasma glucagon concentrations were higher than those of control cats (P < .05), but were not different from those of cats with hepatic lipidosis. Serum insulin concentrations were significantly higher in control cats than in diseased cats (P < .05), but neither serum insulin nor the insulin to glucagon ratio was significantly different among the cats with hepatic disease. The high concentration of NEFAs in cats with hepatic lipidosis suggests that at least 1 factor in the pathogenesis of this syndrome may involve the regulation of hormone sensitive lipase. PMID- 10668813 TI - Treatment of chronic idiopathic large-bowel diarrhea in dogs with a highly digestible diet and soluble fiber: a retrospective review of 37 cases. AB - The medical records of 37 dogs diagnosed with chronic idiopathic large-bowel diarrhea were reviewed. The median age of affected dogs was 6 years. The median body weight was 13.9 kg. The median duration of clinical signs before diagnosis was 32 weeks. Diarrhea usually was intermittent and characterized by increasing frequency, fecal mucus, hematochezia, and tenesmus. Vomiting was common but usually much less frequent and severe than the diarrhea. A variety of stressful factors and abnormal personality traits were identified. CBC and serum biochemistry usually were normal. Fecal examination rarely identified parasites. Rectal cytology specimens were most often normal, but some dogs had increased numbers of neutrophils. The colonic mucosa often was normal during colonoscopy, but decreased numbers of lymphoid follicles were found in some dogs. Histopathologic evaluation found that colonic mucosa was within normal limits. Treatment with soluble fiber (Metamucil) added to a highly digestible diet (Hills i/d) resulted in a very good to excellent response in most dogs. The median initial dosage of Metamucil was 2 tablespoons (2 T) per day. In some dogs, the fiber dosage was reduced or eliminated, or a grocery store brand of dog food was substituted, without causing diarrhea to return. PMID- 10668814 TI - Gastric mucosal lesions in dogs with acute intervertebral disc disease: characterization and effects of omeprazole or misoprostol. AB - We characterized gastric mucosal lesions in dogs with acute degenerative disc disease treated by surgery and corticosteroid administration. The effect of omeprazole and misoprostol on gastric lesions in these dogs was also evaluated. Dogs were randomly assigned to 1 of 2 treatment groups or to the control group. Treatment consisted of omeprazole at 0.7 mg/kg orally once daily, or misoprostol at 2 microg/kg orally 3 times daily. All 3 groups received dexamethasone at 2 mg/kg on day 0, prednisolone at 2 mg/kg on day 1. prednisolone at 1 mg/kg on day 2, and prednisolone at 0.5 mg/kg on all further days (range, 5-6 days). Endoscopic examination was performed on day 0 and 5-6 days later. Four regions of the stomach were qualitatively scored from 1 to 12 based on the presence of submucosal hemorrhage, erosion, or ulceration, with ulceration receiving the highest numerical score. Nineteen of 25 dogs had gastric mucosal lesions at the beginning of the study. No significant difference was found in the gastric lesion score among the 3 groups at the end of the study. Gastric mucosal lesions were concluded to be common in dogs with acute degenerative disc disease treated with corticosteroids. Neither omeprazole nor misoprostol at the doses used was effective in healing or preventing the further development of gastric mucosal lesions. PMID- 10668815 TI - Granulomatous disease associated with Bartonella infection in 2 dogs. AB - Shortly after removal of an engorged tick from the left ear, a 4-year-old Greyhound was referred for evaluation of fever and a rapidly enlarging mass in the region of the left submandibular lymph node. Histopathologic evaluation of the lymph node resulted in a diagnosis of severe granulomatous lymphadenitis. An 11-year-old mixed-breed dog was referred for evaluation of a 6-week history of serous nasal discharge. Histologic examination of a surgical biopsy from a nasal mass indicated multifocal granulomatous inflammation with fibrosis. Serum samples obtained from both dogs were reactive by immunofluorescent assay to Bartonella vinsonii subsp. berkhoffii antigens (reciprocal titers of 128). Although Bartonella organisms were not isolated by lysis centrifugation blood culture, Bartonella DNA was amplified from tissue samples obtained from each dog (lymph node biopsy from dog 1 and nasal biopsy from dog 2) using primers that amplify a portion of the 16S rRNA gene followed by Southern blot hybridization using a genus-specific probe. Additionally, restriction fragment length polymorphism (RFLP) analysis of a Bartonella-specific citrate synthase gene product obtained from dog 2 resulted in a restriction pattern identical to B. vinsonii subsp. berkhoffii. This is the 1st report of granulomatous disease in dogs associated with Bartonella infection. PMID- 10668816 TI - Functional urethral obstruction in 3 dogs: clinical and urethral pressure profile findings. AB - Three dogs with dysuria and urine retention caused by excessive functional urethral resistance are described. All dogs had clinical histories and urologic signs that previously would have been classified as detrusor-urethral dyssynergia. Diagnosis of functional urinary obstruction was established by exclusion of anatomic urinary obstruction and confirmed by urethral pressure profilometry. In 2 cases, multiple pressure deflections recorded in the urethral pressure profile suggested spasm of urethral musculature, whereas in a 3rd dog, abnormally high pressures were recorded along a portion of the proximal urethra. Functional urinary obstruction was associated with prostatitis in 1 dog and with a history of urethral calculi in 1 dog, and no underlying disorder could be identified in the remaining dog. All 3 dogs improved with medical treatments that included alpha adrenergic antagonists. The etiology, diagnosis, and pharmacologic management of functional urinary obstruction are discussed. PMID- 10668817 TI - Seroprevalence of Ehrlichia canis, Ehrlichia equi, and Ehrlichia risticii in sick dogs from North Carolina and Virginia. AB - Ehrlichia canis, E. equi, and E. risticii seroprevalence was determined by microimmunofluorescent antibody testing (IFA) in a sequential population of 1,845 sick dogs admitted during a 1-year period to the North Carolina State University Veterinary Teaching Hospital. A seroreactor was defined by a reciprocal IFA titer of > or =80 to E. canis, E. equi, or E. risticii antigens. Of the 48 IFA seroreactors, 44 dogs were seroreactive to E. canis, 21 to E. equi, and 0 to E. risticii. Seventeen dogs reacted to both E. canis and E. equi antigens. There was concordance of E. canis IFA and western immunoblot (WI) test results for 36/44 dogs. Because of cross-reactivity of E. canis sera with E. equi antigens, WI was of less utility to confirm E. equi exposure. After elimination of E. canis seroreactors, there was concordance of 2/4 E. equi IFA and WI test results. Based upon a retrospective review of medical records, ehrlichiosis was diagnosed in 10/48 (21%) IFA seroreactive dogs, 9 of which were confirmed positive by WI. Of the remaining 38 IFA seroreactors, 29 also were confirmed by E. canis or E. equi WI. These results indicate that (1) ehrlichiosis was not diagnosed in the majority of serologically confirmed cases, (2) based upon E. canis and E. equi WI analysis, IFA testing was not specific (21% false positive), (3) E. canis sera cross-react with E. equi antigens, and (4) serologic evidence of E. risticii infection was lacking in the dog population studied. PMID- 10668818 TI - Prothrombin, activated partial thromboplastin, and proteins induced by vitamin K absence or antagonists clotting times in 20 hyperthyroid cats before and after methimazole treatment. AB - The effect of daily doses of 5-15 mg of methimazole on the platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), and proteins induced by vitamin K absence or antagonists (PIVKA) clotting time in 20 hyperthyroid cats was determined. No significant (P > .05) difference was found in median platelet count. PT, APTT, or PIVKA clotting time before treatment compared to median values at 2-6 weeks or > or =7-12 weeks of methimazole treatment. No cat had a prolonged APTT at any time. At 2-6 weeks of methimazole treatment, 1 cat each developed thrombocytopenia or prolonged PIVKA clotting time despite initially normal values. Three cats had abnormal coagulation tests (prolonged PT [n = 1] and PIVKA clotting time [n = 3]) before treatment that fluctuated during treatment. Excluding the 3 cats that had abnormal PIVKA clotting time before treatment, prolonged PIVKA clotting time developed in 6% (1/17; 95% confidence interval, 0-28%) cats treated with methimazole for 2-6 weeks. Seemingly. doses of methimazole commonly used to treat hyperthyroidism in cats do not cause alteration in PT and APTT, and only rarely prolong PIVKA clotting time. Nevertheless, abnormal PIVKA clotting time may explain bleeding tendencies unassociated with thrombocytopenia in methimazole-treated hyperthyroid cats. PMID- 10668819 TI - Food hypersensitivity reactions in Soft Coated Wheaten Terriers with protein losing enteropathy or protein-losing nephropathy or both: gastroscopic food sensitivity testing, dietary provocation, and fecal immunoglobulin E. AB - The purpose of this study was to evaluate Soft Coated Wheaten Terriers (SCWTs) affected with protein-losing enteropathy (PLE) or protein-losing nephropathy (PLN) or both for allergy to food. We performed gastroscopic food-sensitivity testing, a provocative dietary trial, and measurement of fecal immunoglobulin E (IgE) in 6 SCWTs affected with PLE or PLN or both. Positive gastroscopic food sensitivity test reactions were noted in 5 of 6 dogs. Positive reactions were found to milk in 4 dogs, to lamb in 2 dogs, and to wheat and chicken each in 1 dog. Adverse reactions to food (diarrhea, vomiting, or pruritus) were detected in all 6 dogs during the provocative dietary trial. Adverse reactions were found to corn in 5 dogs, to tofu in 3 dogs, to cottage cheese in 2 dogs, to milk in 2 dogs, to farina cream of wheat in 2 dogs, and to lamb in 2 dogs. Serum albumin concentrations significantly decreased and fecal alpha1-protease inhibitor concentration significantly increased 4 days after the provocative trial when compared with baseline values. Antigen-specific fecal IgE varied throughout the provocative trial, with peak levels following ingestion of test meals. We conclude that food hypersensitivities are present in SCWTs affected with the syndrome of PLE/PLN. Mild inflammatory bowel disease was already established in the 6 SCWTs of this report at the time of study, making it impossible to determine if food allergies were the cause or result of the enteric disease. PMID- 10668820 TI - Familial protein-losing enteropathy and protein-losing nephropathy in Soft Coated Wheaten Terriers: 222 cases (1983-1997). AB - Records and pedigrees of Soft Coated Wheaten Terriers (SCWT) with protein-losing enteropathy (PLE) or protein-losing nephropathy (PLN) were studied retrospectively. Criteria for inclusion were defined based on analysis of blood (panhypoproteinemia for PLE, hypoalbuminemia for PLN) and urine (proteinuria for PLN) and histopathologic examination of tissue. Two hundred twenty-two affected dogs (female:male ratio = 1.6, P < .001) were clinically identified. Dogs were diagnosed with PLE earlier (P < .005; mean +/- SD age: 4.7+/-2.6 years, n = 76) than with PLN (6.3+/-2.0 years, n = 84) or with both diseases (5.9+/-2.2 years, n = 62). Clinical signs included vomiting, diarrhea, weight loss, pleural and peritoneal effusions, and less commonly thromboembolic disease. Dogs with PLE generally had panhypoproteinemia and hypocholesterolemia; intestinal lesions included inflammatory bowel disease, dilated lymphatics, and lipogranulomatous lymphangitis. Dogs with PLN generally had hypoalbuminemia, proteinuria, hypercholesterolemia, and azotemia; renal lesions typically showed chronic glomerulonephritis/glomerulosclerosis, and less commonly endstage renal disease. Dogs with combined PLE/PLN had intermediate mean values (P < .001) for serum total protein, albumin, globulin, and cholesterol but had a higher mean urine protein:creatinine ratio than did PLN dogs (P < .05); intestinal and renal lesions in these dogs were similar to those in the other groups. Two dogs had incidental mild renal dysplasia. Pedigree analysis from 188 dogs demonstrated a common male ancestor, although the mode of inheritance is unknown. Both PLE and PLN are common diseases in this small breed population. The prognosis is poor. Compared with previously reported intestinal and renal diseases in dogs, a new, distinctive familial predisposition for both PLE and PLN has been recognized in the SCWT breed. PMID- 10668821 TI - Vincristine impairs platelet aggregation in dogs with lymphoma. AB - Platelet aggregation before and after administration of 0.5 mg/m2 of vincristine (VCR) was evaluated in 7 dogs with spontaneously occuring lymphoma. Aggregation on platelet-rich plasma separated from blood collected in 3.8% sodium citrate was performed using adenosine diphosphate (ADP), arachidonic acid (AA), and collagen (COL) as agonists. The slope for aggregation in response to ADP was significantly lower after administration of VCR (P = .032). Maximal aggregation after administration of VCR was significantly lower in response to ADP, COL, and AA (P = .03, P = .04, and P = .03, respectively). PMID- 10668822 TI - Clinical evaluation of methoximorpholino-doxorubicin (FCE 23762) in dogs with spontaneous malignancies. AB - We conducted a clinical evaluation of FCE 23762, a methoxymorpholino analog of doxorubicin, in 48 dogs with metastatic, nonresectable, or chemotherapy-resistant spontaneous malignancies at an initial dosage of 50-60 microg/kg IV every 3 weeks. Clinical evidence of toxicity was minimal; 6 dogs developed grades I, II, and III hematologic toxicities after the 1st treatment, and 1 dog developed grade II gastrointestinal toxicity. One dog became pancytopenic 4 months after discontinuation of FCE 23762. No other adverse effects were noted. Partial or complete remissions were observed in 32% of the dogs. Responses were observed both in previously untreated dogs and in those that had received prior chemotherapy, including doxorubicin. FCE 23762 is a promising new antineoplastic agent that can be used safely in dogs with cancer; doses higher than those used in this study may be used eventually in practice. PMID- 10668823 TI - Hemodynamic and electrocardiographic effects of graded doses of amiodarone in healthy dogs anesthetized with morphine/alpha chloralose. AB - This research was designed to study acute hemodynamic and electrocardiographic effects of amiodarone and to determine an IV dose of amiodarone that minimally affects left ventricular function and does not increase the tendency for ventricular arrhythmia. PMID- 10668825 TI - Necrosis of hippocampus and piriform lobe in 38 domestic cats with seizures: a retrospective study on clinical and pathologic findings. AB - The clinical records of 38 cats (1985-1995) with a neuropathologically confirmed diagnosis of necrosis of the hippocampus and occasionally the lobus piriformis were evaluated retrospectively. There was no sex or breed predisposition. Most cats were between 1 and 6 years of age (mean age 35 months) and had either generalized or complex-partial seizures of acute onset and rapid progression. The seizures had a tendency to become recurrent and to present as clusters or even status epilepticus later in the course of the disease. Fourteen cats died spontaneously, and 24 were euthanized. Histopathologic examination revealed bilateral lesions restricted to the hippocampus and occasionally the lobus piriformis. The lesions seemed to reflect different stages of the disease and consisted of acute neuronal degeneration to complete malacia, affecting mainly the layer of the large pyramidal cells but sometimes also the neurons of the dentate gyrus and the piriform lobe. The clinical, neuropathologic, and epidemiologic findings suggest that the seizures in these cats were triggered by primary structural brain damage, perhaps resulting from excitotoxicity. The cause remains unknown, but epidemiologic analysis suggests an environmental factor, probably a toxin. PMID- 10668824 TI - Hepatopathy in 4 dogs treated with amiodarone. AB - Amiodarone is a class III antiarrhythmic drug used in dogs with dilated cardiomyopathy and ventricular tachyarrhythmias. Hepatopathy is one of the more commonly reported adverse effects of amiodarone use in people. We describe 4 dogs that developed hepatopathy associated with amiodarone administration; 2 dogs also developed neutropenia. Three dogs had clinical signs of anorexia and lethargy; 1 did not show signs until impaired liver function had developed. Clinical signs or biochemical abnormalities developed 1.5-8 months after amiodarone treatment was started. Clinical signs resolved within 2 weeks of discontinuing amiodarone, but biochemical abnormalities did not resolve for 6-8 weeks. The delay between onset of liver disease and overt clinical signs suggests that serial evaluation of liver enzyme activities following amiodarone use in does is important. PMID- 10668826 TI - Disseminated hemangiosarcoma in the horse: 35 cases. AB - Thirty-five cases of disseminated hemangiosarcoma (21 clinical cases and 14 previously reported cases) were reviewed to describe the disease in horses. Hemangiosarcoma occurred in mature, particularly middle-aged horses, with no apparent sex predilection. Thoroughbreds seemed to be overrepresented (13 cases) but a true breed predilection could not be established. The respiratory and musculoskeletal systems were most commonly affected and presenting complaints included dyspnea (26%), subcutaneous or muscular swelling (24%), epistaxis (17%), and lameness (12%). Heart and respiratory rates were usually increased and mucous membrane color was frequently pale or icteric. Capillary refill time and rectal temperature were often normal. Anemia (88%), neutrophilic leukocytosis (62%), and thrombocytopenia (48%) were common. Examination of tissue samples collected by fine-needle aspirate or biopsy established an antemortem diagnosis in 4 horses. The diagnosis was made during postmortem examination in the remaining 31 horses. The lung and pleura (77%), skeletal muscle (46%), and spleen (43%) were most commonly affected. A primary site of tumor involvement could be identified in 22 horses. Hemangiosarcoma should be included as a differential diagnosis for horses with evidence of hemorrhage into body cavities, skeletal muscle, or subcutaneous locations. PMID- 10668827 TI - Acute B-cell lymphoblastic leukemia with meningeal metastasis causing primary neurologic dysfunction in a dog. PMID- 10668828 TI - The market pushes education from ward to office, from acute to chronic illness and prevention: will case method teaching-learning change? PMID- 10668829 TI - Hypertension in the elderly: can we improve results of therapy? AB - Awareness of and therapy for hypertension in the United States have been increasing in older patients. Despite this improvement, hypertension continues to be poorly controlled in this patient population. The control rate, defined as systolic blood pressure less than 140 mm Hg and diastolic blood pressure less than 90 mm Hg, is surprisingly low for older patients, despite abundant data documenting the reduction of cardiovascular events by treating both systolic diastolic and isolated systolic hypertension. Comorbid diseases and physiological alterations in the elderly, including reduced myocardial contractility, renal function, total body water, baroreceptor responsiveness, and cognitive function, must be considered, but in general these have not limited the effectiveness of antihypertensive drug therapy. PMID- 10668830 TI - The role of spiral volumetric computed tomography in the diagnosis of pulmonary embolism. AB - To evaluate the evidence for the use of spiral volumetric computed tomography (SVCT) in the diagnosis of acute pulmonary embolism (PE), the 11 English-language studies published through July 1998 that compared SVCT with a reference standard for PE were systematically reviewed. Among the reviewed studies, methodological problems were common. Only 5 of these studies fulfilled 5 of 11 basic standards addressing important issues in diagnostic test research. The reported sensitivities of SVCT compared with pulmonary angiography varied widely (64% 93%), which was likely the result of differences in study populations. Spiral volumetric computed tomography may be relatively sensitive and specific for diagnosing central pulmonary artery PEs, but it is insensitive for diagnosing subsegmental clots. Spiral volumetric computed tomography may have a role as a "rule-in" test for large central emboli, but additional research is required to establish its place in clinical practice. PMID- 10668831 TI - Improving preventive care by prompting physicians. AB - OBJECTIVES: To assess the impact of prompting physicians on health maintenance, answer questions regarding the mode of delivery, and identify opportunities and limitations of this information intervention. METHODS: Systematic electronic and manual searches (January 1, 1966, to December 31, 1996) were conducted to identify clinical trial reports on prompting clinicians. Three eligibility criteria were applied: (1) randomized controlled clinical trial, (2) clinician prompt, alert, or reminder in the study group and no similar intervention in the control group, and (3) measurement of the intervention effect on the frequency of preventive care procedures. Data were abstracted by independent reviewers using a standardized abstraction form, and quality of methodology was scored. A series of meta-analyses on triggering clinical actions was performed using the random effects method. The statistical analyses included 33 eligible studies, which involved 1547 clinicians and 54 693 patients. RESULTS: Overall, prompting can significantly increase preventive care performance by 13.1% (95% confidence interval [CI], 10.5%-15.6%). However, the effect ranges from 5.8% (95% CI, 1.5% 10.1%) for Papanicolaou smear to 18.3% (95% CI, 11.6%-25.1%) for influenza vaccination. The effect is not cumulative, and the length of intervention period did not show correlation with effect size (R = -0.015, P = .47). Academic affiliation, ratio of residents, and technique of delivery did not have a significant impact on the clinical effect of prompting. CONCLUSIONS: Dependable performance improvement in preventive care can be accomplished through prompting physicians. Vigorous application of this simple and effective information intervention could save thousands of lives annually. Health care organizations could effectively use prompts, alerts, or reminders to provide information to clinicians when patient care decisions are made. PMID- 10668832 TI - Predictive value of compression ultrasonography for deep vein thrombosis in symptomatic outpatients: clinical implications of the site of vein noncompressibility. AB - BACKGROUND: Compression ultrasonography has a high negative predictive value for deep vein thrombosis in symptomatic outpatients. Limited data are available on factors influencing positive predictive value. The objective of this study was to evaluate the positive predictive value of compression ultrasonography according to the anatomic site of vein noncompressibility. METHODS: We performed a prospective cohort study of 756 consecutive outpatients with suspected first episode deep vein thrombosis. Compression ultrasonography was performed at the initial visit: results were abnormal if a noncompressible segment was identified or normal if all segments were fully compressible. Venography was performed in patients with abnormal compression ultrasonography results. Positive predictive value was determined according to the site of noncompressibility: common femoral vein only, popliteal vein only, or both sites. Venography was the reference standard for the presence of deep vein thrombosis. RESULTS: Positive predictive value was 16.7% (95% confidence interval, 0.4%-64.1%) for noncompressibility isolated to the common femoral vein compared with 91.3% (95% confidence interval, 72.0%-98.9%) for the popliteal vein only and 94.4% (95% confidence interval, 72.7%-99.9%) for both sites (P<.001). Of 15 patients with isolated noncompressibility of the common femoral vein, 8 (53%) had pelvic neoplasm or abscess compared with 2 (5%) of 42 with noncompressibility of the popliteal vein only and 6 (13%) of 47 with noncompressibility of both sites (P<.001). CONCLUSIONS: The positive predictive value of noncompressibility isolated to the common femoral vein is too low to be used alone as the diagnostic end point for giving anticoagulant therapy. Noncompressibility isolated to the common femoral vein is a diagnostic marker for pelvic disease. PMID- 10668833 TI - Prevalence and content analysis of guidelines on handling requests for euthanasia or assisted suicide in Dutch nursing homes. AB - BACKGROUND: The growing number of requests for euthanasia or assisted suicide (EAS) makes it imperative for health care institutions, such as nursing homes, to have written guidelines on how to handle requests for EAS. The objective of this study was to determine the prevalence of EAS guidelines in Dutch nursing homes and to analyze the content. METHODS: Directors of patient care in 324 Dutch nursing homes were asked, by means of a mailed short list of questions, if they had an institutional guideline on EAS and, if so, to provide a copy. Guidelines were analyzed according to a structured list of items based on current jurisprudence, model documents, and opinions of experts. RESULTS: Of the 324 directors, 313 (97%) responded. In 58% of the nursing homes that responded, there existed written guidelines for EAS. Of those guidelines, 74% concerned EAS; in 26%, EAS was integrated in a guideline on terminal care. Of the guidelines, 165 (90%) were based on the policy that EAS is acceptable under specific conditions, and 18 (10%) banned EAS completely. Of the first-mentioned guidelines, 81% described one or more procedures for in-principle objections. In 65% of these guidelines, all official requirements for prudent practice were described. CONCLUSIONS: Despite the rapidly growing number of nursing-home guidelines on EAS and the existence of model documents, there is still considerable variation in the guidelines, and they can be improved in many aspects. A basic prerequisite is that the guidelines include all the official requirements for prudent practice. PMID- 10668834 TI - Ten-year trends in hospital care for congestive heart failure: improved outcomes and increased use of resources. AB - BACKGROUND: Scarce data are available on long-term trends in hospital mortality, length of stay (LOS), and costs in congestive heart failure (CHF). OBJECTIVE: To assess 10-year trends in the outcomes of patients hospitalized with CHF. METHODS: We studied all 6676 patients with a primary discharge diagnosis of CHF hospitalized from January 1, 1986, through July 31, 1996, at an academic tertiary care center. Hospital mortality, LOS, and costs were adjusted for sociodemographic characteristics, comorbidities, invasive procedures, hospital disposition, and LOS where appropriate. RESULTS: The mean (+/- SD) age of patients was 70+/-13 years; 54.1% were male; 87.0% were white. There was a significant increasing trend in heart failure severity as assessed by a CHF specific risk-adjustment index. The proportion of patients who underwent invasive procedures (e.g., cardiac catheterization, coronary angioplasty, coronary artery bypass surgery, defibrillator and pacemaker implantation) was significantly higher in the 1994-1996 period. The standardized mortality ratio (observed mortality/predicted mortality) progressively fell during the study period. Compared with patients admitted before 1991, those admitted after 1991 had a 24% lower observed than predicted mortality. Adjusted LOS exhibited a downward trend, ie, 7.7 days in 1986-1987 to 5.6 days in 1994-1996 (P<.001). Unadjusted cost peaked during 1992-1993 and declined thereafter. Adjusted costs in 1994-1996 were not significantly different from those in 1990-1991. CONCLUSIONS: After risk adjustment for sociodemographic characteristics, comorbidities, and disease severity, a significant decrease in in-hospital mortality was observed during the study decade. This decline in hospital mortality occurred in parallel with decreasing LOS and increasing use of cardiac procedures and costs. PMID- 10668835 TI - Prevention of venous thromboembolism: adherence to the 1995 American College of Chest Physicians consensus guidelines for surgical patients. AB - BACKGROUND: The American College of Chest Physicians addressed the dilemma of identifying optimal therapy for venous thromboembolism (VTE) prophylaxis and published their Fourth Consensus Conference on Antithrombotic Therapy in 1995, with recommendations for prophylactic therapy. Despite these recommendations, appropriate VTE prophylactic therapy is underused. OBJECTIVES: To examine routine practices in the prevention of VTE in high-risk surgical patients and to determine the extent of adoption of grade A prophylactic therapies as recommended by the American College of Chest Physicians. METHODS: Retrospective medical record review in 10 teaching or community-based hospitals located in the United States. Medical charts of 1907 patients were randomly selected for review from the population of patients who underwent high-risk major abdominal surgery, total hip replacement, hip fracture repair, or total knee replacement between January 1, 1996, and February 28, 1997. RESULTS: Of 1907 patients, VTE prophylaxis was used in 89.3%; use was 93.7% in each of the 3 orthopedic surgery groups and 75.2% in the high-risk major abdominal surgery group. The percentage of patients receiving grade A therapy was highest in the hip replacement group (84.3%) vs. the other groups (knee replacement, 75.9%; hip fracture repair, 45.2%; abdominal surgery, 50.3%). CONCLUSIONS: The use of grade A prophylaxis was related to the type of surgery, with the highest use seen in total hip replacement and the lowest in hip fracture repair. One in 4 patients who underwent high-risk major abdominal surgeries failed to receive any form of VTE prophylaxis. Publication of consensus statements alone may be insufficient to ensure the incorporation of important new clinical information into routine practice. PMID- 10668836 TI - Sex bias and underutilization of lipid-lowering therapy in patients with coronary artery disease at academic medical centers in the United States and Canada. Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial (PREVENT) Investigators. AB - BACKGROUND: The efficacy of lipid-lowering therapy (LLT) has been well established for patients with preexisting coronary artery disease (CAD). However, limited information is available assessing the extent to which these medications are prescribed in academic medical centers. METHODS: The use of LLT for patients with CAD was prospectively evaluated in 825 men and women who were recruited from 16 academic medical centers in the United States and Canada to participate in the Prospective Evaluation of the Vascular Events of Norvasc Trial (PREVENT). The assessment of LLT use during the 3-year trial was evaluated in patients receiving amlodipine therapy and placebo; levels of low-density lipoprotein cholesterol (LDL-C) were used to assess the impact of LLT. RESULTS: Despite a baseline prevalence of LLT in 42% of men (38% in 1994), half of the patients had high levels of LDL-C (>3.36 mmol [>130 mg/dL]). During the subsequent 3 years, the prevalence of elevated LDL-C levels dropped in men (29%) but remained stagnant in women (48%). These changes were associated with increased LLT in men (55%) but not in women (35%) (P = .04). In 1994, the LDL-C target goal (<2.59 mmol/L [<100 mg/dL]) was attained in 17% of men and 6% of women (P = .006). At study completion in 1997, the LDL-C target goal was achieved in 31% of men and only 12% of women (P = .001). CONCLUSIONS: This study highlights the relatively low treatment rates of hyperlipidemia among patients with CAD overall and women in particular who were participating in a clinical trial at academic medical centers in the United States and Canada. Because LLT has been proven to reduce future cardiovascular events, these results suggest that more intensive efforts should be promoted in order to maximize CAD reduction. PMID- 10668837 TI - Angiotensin-converting enzyme inhibitors, calcium channel blockers, and breast cancer. AB - BACKGROUND: The use of angiotensin-converting enzyme (ACE) inhibitors has been linked to a decreased risk of developing cancer, and longer-term use of calcium channel blockers (CCBs) has been associated with an increased risk of developing cancer in general and breast cancer in particular. METHODS: Using data from the General Practice Research Database, we conducted a large case-control analysis. Previous exposure to ACE inhibitors, CCBs, and beta-blockers was compared between 3706 postmenopausal women who were diagnosed with incident breast cancer between 1992 and 1997 and 14155 matched-control women. RESULTS: Compared with nonusers of antihypertensive drugs, women who used ACE inhibitors (odds ratio [OR], 1.0; 95% confidence interval [CI], 0.7-1.5), CCBs (OR, 0.9; 95% CI, 0.7-1.2), or beta blockers (OR, 1.0; 95% CI, 0.8-1.2) for 5 or more years were not at an increased or decreased risk of developing breast cancer (adjusted for smoking and body mass index [calculated as weight in kilograms divided by the square of height in meters]). The risk of breast cancer did not differ between users of different ACE inhibitors or different CCBs (dihydropyridines, diltiazem hydrochloride, and verapamil hydrochloride) or between users of short-acting (OR, 1.0; 95% CI, 0.7 1.4) or sustained-release (OR, 1.0; 95% CI, 0.8-1.3) nifedipine preparations. CONCLUSION: The findings of this large case-control analysis do not support the hypothesis that longer-term use of ACE inhibitors or CCBs affects the risk of developing breast cancer. PMID- 10668838 TI - A nationwide study of decisions to forego life-prolonging treatment in Dutch medical practice. AB - BACKGROUND: Decisions to withhold or withdraw life-prolonging treatment in terminally ill patients are common in some areas of medical practice. Information about the frequency and background of these decisions is generally limited to specific clinical settings. This article describes the practice of withholding or withdrawing life-prolonging treatment in the Netherlands. METHODS: Questionnaires were sent to the attending physicians of a stratified sample of 6060 of all 43002 cases of death in the Netherlands from August 1 through November 30, 1995. The questions concerned the treatments foregone, the patient characteristics, and the decision-making process. The response rate was 77%. RESULTS: A nontreatment decision was made in 30% (95% confidence interval, 28%-31%) of all deaths in the Netherlands in 1995; this is an increase compared with 28% (95% confidence interval, 26%-29%) in 1990; in 20% of all deaths, this decision was the most important end-of-life decision. Artificial nutrition or hydration and antibiotics were the treatments most frequently foregone, each accounting for 25% of cases in which a nontreatment decision was made. Nursing-home physicians withheld or withdrew treatment more often than clinical specialists or general practitioners in 52%, 35%, and 17% of all deaths they were involved with, respectively. Of the patients in whom a nontreatment decision was the most important end-of-life decision, 26% were competent; of those, 93% were involved in the decision making. In 17% of patients, the nontreatment decision was made without being discussed with the patient or the patient's relatives and without knowledge of the patient's wishes. Life was shortened by an estimated 24 hours or less in 42% and 1 month or more in 8% of patients. CONCLUSIONS: Decisions to forego life prolonging treatment are frequently made end-of-life decisions in the Netherlands and may be increasing. Most of these decisions do not involve high-technology treatments, and the consequences, in terms of shortening of life, are relatively small. PMID- 10668839 TI - Marked declines in human immunodeficiency virus-related mortality in Chicago in women, African Americans, Hispanics, young adults, and injection drug users, from 1995 through 1997. AB - BACKGROUND: Declines in human immunodeficiency virus (HIV)-related mortality between 1995 and 1996 were seen across the United States but were small to nonexistent among people in marginalized sectors who are most likely to contract HIV and die of its effects. No comprehensive analysis describing HIV-related mortality in 1997 was available. OBJECTIVE: To describe Chicago's HIV-related mortality trends up to and including 1997, with specific attention focused on marginalized populations. METHODS: An analysis of cross-sectional HIV-related mortality data with emphasis on the years 1995 through 1997 was conducted for Chicago, Ill. Numbers, proportions, and rates of declines in HIV-related deaths were examined for the city as a whole and also among those diagnosed at Cook County Hospital, as a proxy for people with very low socioeconomic status. RESULTS: Between 1995 and 1996 there was an overall decline of 19% in HIV-related mortality in Chicago but small or no declines among women, African Americans, Hispanics, injection drug users, and people aged 20 to 29 years and more than 50 years. Between 1995 and 1997 there was an overall decline of 61%. At that time the declines were spread more evenly across diverse groups. There were almost no significant differences between the declines for these groups at Cook County Hospital and in the rest of Chicago. CONCLUSIONS: The HIV-related mortality has fallen dramatically in Chicago since 1995, the year of its maximum. During 1997, declines were seen among all groups. Declines were also seen among the most disenfranchised of the city. Access to care and the new combination therapies are apparently sustaining life for many in Chicago. PMID- 10668840 TI - Autopsy consent practice at US teaching hospitals: results of a national survey. AB - BACKGROUND: Autopsy rates continue to fall despite the enduring benefit of the procedure to families and medical science, yet there are few data about the consent process itself. OBJECTIVE: To evaluate the current practice of obtaining autopsy consent, by assessing the consent forms currently in use, the knowledge and attitudes of chief residents on the procedure, and the expert opinion of pathologists in those institutions. DESIGN: Cross-sectional survey. SETTINGS AND PARTICIPANTS: One hundred twenty-seven US teaching hospitals. RESULTS: Of all autopsy consent forms we surveyed, 84.7% contained 7 of 10 elements recommended by the College of American Pathologists. Only 7.1% of institutions supplied educational materials for the physician, as recommended by the College of American Pathologists. Overall, 50.1% of chief residents reported deficiencies in their knowledge of the autopsy procedure. Correspondingly, greater than 74.5% felt that educational materials would be beneficial for physicians and the family. Finally, 93.3% of chief residents believed that a limited autopsy should be offered to families, while 68 (90%) of 76 pathologists at these institutions believed that limited autopsies are an unsatisfactory alternative to the complete procedure. CONCLUSIONS: Chief residents at US teaching hospitals reported substantial deficiencies in their knowledge about autopsy and desire more training on the consent process. Autopsy consent forms are often lacking information that might help physicians and families in making an educated choice about autopsy. Teaching institutions need to reevaluate the training for the autopsy consent practice. PMID- 10668841 TI - Hypercoagulable states in primary upper-extremity deep vein thrombosis. AB - BACKGROUND: There are very few data on the prevalence of coagulation abnormalities in primary deep vein thrombosis of the upper limbs. OBJECTIVE: To determine if coagulation abnormalities play a role in effort-related and/or idiopathic (non-effort-related) upper-extremity deep vein thrombosis (UEDVT). METHODS: Fifty-one consecutive patients (21 men and 30 women) who had effort related (n = 20) or idiopathic (n = 31) UEDVT over an 18-year period (median age at diagnosis, 32 years; age range, 15-86 years) were routinely reexamined. Plasma was screened for antithrombin, protein C, and protein S deficiencies and for antiphospholipid antibodies (lupus anticoagulant and anticardiolipin antibodies). The DNA was screened for factor V Leiden and for prothrombin gene G20210A mutations. RESULTS: The median age (35 vs. 28 years), the proportion of women (81% [25/31] vs. 25% [5/201), the proportion of patients with a personal and/or family history of thromboembolism (42% [13/31] vs. 15% [3/20]), and the proportion of patients with at least 1 coagulation abnormality (42% [13/31] vs. 15% [3/20]) were higher in the idiopathic UEDVT group than in the effort-related UEDVT group. The odds ratio of having a coagulation abnormality was 4.09 (95% confidence interval, 0.99-16.78; P = .06) in the idiopathic UEDVT group compared with the effort-related UEDVT group. CONCLUSION: Hypercoagulable states appear to play a significant role in idiopathic but not in effort-related UEDVT. PMID- 10668842 TI - Fulminant non--A-G viral hepatitis leading to liver transplantation. AB - BACKGROUND: All hepatotropic viruses are known to cause fulminant hepatic failure (FHF). However, in 30% to 40% of patients with FHF, the precise cause remains unknown. We aimed to better define this subgroup. METHODS: We evaluated the clinical course and outcome of 7 patients admitted during a 22-month period with fulminant viral hepatitis leading to liver transplantation; none had serologic or molecular evidence of hepatitis A, B, C, D, E, or G viral infection, thus the term non-A-G viral hepatitis. All known etiologies of FHF were excluded. RESULTS: All patients had prodromal symptoms suggestive of viral causes. Mean age was 30 years. The interval between onset of jaundice and appearance of encephalopathy was 23 days (range, 4-50 days). Five patients had grade III/IV encephalopathy. Serum alanine aminotransferase levels showed a single peak of activity. The duration between first symptoms and liver transplantation was 28 days (range, 12 71 days). Results of histological study of the explanted liver showed submassive (4 patients) or massive (3 patients) hepatocyte necrosis. In all patients, results of polymerase chain reaction analysis did not detect hepatitis B virus DNA, hepatitis C virus RNA, or hepatitis G virus RNA in the explanted liver. After transplantation, 2 patients showed (6 months later) increased liver enzyme levels of undetermined cause, and results of a liver biopsy showed mild lobular hepatitis; 1 patient had lymphoproliferative disorder (Epstein-Barr virus originated); and 1 patient, aplastic anemia, which is known to be associated with seronegative viral hepatitis. The latter patient died, whereas the other 6 patients are alive (survival rate, 86%). CONCLUSIONS: Our patients with non-A-G viral hepatitis had a severe acute onset with progressive FHF requiring liver transplantation. There is some suggestion of recurrent viral disease after transplantation implicating other unknown viruses in the etiology. PMID- 10668843 TI - Successful treatment with combination therapy of cyclophosphamide and cyclosporin for late recurrence of Wegener granulomatosis. PMID- 10668844 TI - Aspirin in the treatment of type 2 diabetes. PMID- 10668845 TI - Virologic response associated with a change in protease inhibitor therapy. PMID- 10668846 TI - The lipid-lowering effect of lean meat diets falls far short of that of vegetarian diets. PMID- 10668847 TI - Increased bioavailability of oral melatonin after fluvoxamine coadministration. AB - BACKGROUND: Fluvoxamine, a selective serotonin reuptake inhibitor, is known to elevate melatonin serum concentrations. It has not been clear whether these effects might be attributed to an increased melatonin production or to an decreased elimination of melatonin. The latter hypothesis was tested by this study. METHODS: Five healthy male volunteers (one CYP2D6 poor metabolizer) received 5 mg melatonin either with or without coadministration of 50 mg fluvoxamine. Serum concentrations of melatonin and fluvoxamine were assessed from 0 to 28 hours after melatonin intake. RESULTS: Coadministration of fluvoxamine, on average, led to an 17-fold higher (P < .05) area under concentration-time curve (AUC) and a 12-fold higher (P < .01) serum peak concentration (Cmax) of melatonin. The terminal elimination half-life was not significantly affected. The AUC and Cmax of fluvoxamine were about three times higher and the half-life was about two times higher in the poor metabolizer. There was a correlation (r = 0.63; P < .01) between the melatonin and fluvoxamine serum concentrations. The poor metabolizer was found to have a more pronounced and longer-lasting effect of fluvoxamine on the pharmacokinetics of melatonin. CONCLUSION: This study showed an increase in the bioavailability of oral melatonin by coadministration of fluvoxamine. The effects of fluvoxamine on the melatonin serum concentrations in patients with depression might therefore be caused by inhibition of the elimination of melatonin and not attributable to an increased production of melatonin. PMID- 10668849 TI - Gentamicin effects on urinary electrolyte excretion in healthy subjects. AB - BACKGROUND: Symptomatic hypomagnesemia, hypocalcemia, and hypokalemia caused by renal electrolyte wasting occasionally develop in patients treated with aminoglycosides. This phenomenon has been attributed to aminoglycoside tubular injury. However, rats administered a single dose of gentamicin show immediate dose-related calcium and magnesium renal wasting without sodium or potassium wasting days before other evidence of tubular dysfunction or structural injury can be shown. The mechanism is undefined but transient and is not dependent on the presence of parathyroid hormone. OBJECTIVE: To determine whether gentamicin administration to humans causes renal electrolyte wasting. DESIGN: Five healthy volunteers ingested a 400-mg calcium, 100-mEq sodium diet for 1 week before the study. After a 90-minute baseline period, 5 mg/kg gentamicin was administered intravenously over 30 minutes. Urine and serum were collected for 5 hours after gentamicin administration. RESULTS: Peak serum gentamicin levels ranged from 12.8 to 20.6 microg/mL. There was no change in serum electrolytes. The urinary fractional calcium excretion rose from a baseline of 1.8% +/- 0.5% to 6.8% +/- 1.4% (P < .01), and the magnesium fractional excretion rose from 3.4% +/- 0.8% to 11.8% +/- 6.4% (P = .03). These effects were transient. Gentamicin caused no change in renal excretion of sodium, potassium, or phosphate. CONCLUSIONS: Gentamicin administered at the standard clinical dose causes immediate and transient renal calcium and magnesium wasting in normal humans. The mechanism of gentamicin-associated urinary magnesium wasting and calciuria is undefined. However, the pattern of electrolyte excretion after gentamicin administration suggests that the site of action of these gentamicin effects is the distal convoluted tubule. PMID- 10668848 TI - Pharmacokinetics of repaglinide in subjects with renal impairment. AB - OBJECTIVE: To evaluate the effect of renal impairment and renal failure on the pharmacokinetics and safety of repaglinide. METHODS: We conducted a phase I, multicenter, parallel-group, pharmacokinetic comparison trial with single and multiple doses of repaglinide in subjects with various degrees of renal impairment. Subjects with normal renal function (n = 6) and subjects with renal impairment (mild to moderate, n = 6; severe, n = 6) received treatment with 2 mg repaglinide for 7 days. Subjects in the hemodialysis group (n = 6) received two single doses of 2 mg repaglinide separated by a 7- to 14-day washout period. All subjects had repaglinide serum pharmacokinetic profiles measured for the first and last doses administered. Serum steady-state levels, urine levels, and dialysate levels were also measured. RESULTS: Pharmacokinetic parameters did not show significant changes after single or multiple doses of repaglinide, although the elimination rate constant in the group with severe renal impairment decreased after 1 week of treatment. Subjects with severe impairment had significantly higher area under the curve values after single and multiple doses of repaglinide than subjects with normal renal function. No significant differences in values for maximum serum concentration or time to reach maximum concentration were detected between subjects with renal impairment and those with normal renal function. Hemodialysis did not significantly affect repaglinide clearance. CONCLUSIONS: Repaglinide was safe and well tolerated in subjects with varying degrees of renal impairment. Although adjustment of starting doses of repaglinide is not necessary for renal impairment or renal failure, severe impairment may require more care when upward adjustments of dosage are made. PMID- 10668850 TI - Local venous response to N-desethylamiodarone in humans. AB - OBJECTIVE: Amiodarone, a class III antiarrhythmic agent, is a potent vasodilator in vivo. Its main metabolite, N-desethylamiodarone, contributes to the antiarrhythmic action of amiodarone after long-term treatment. It is unknown whether N-desethylamiodarone has acute vascular effects. The aim of this study was to explore the mechanism of action of N-desethylamiodarone in human hand veins. METHODS: The dorsal hand vein compliance technique was applied in 36 healthy male volunteers. In hand veins preconstricted with the alpha1-adrenergic receptor agonist phenylephrine or prostaglandin F2alpha, N-desethylamiodarone and an inhibitor of nitric oxide formation (N(G)-monomethyl-L-arginine, L-NMMA) were infused in the presence or absence of a cyclooxygenase inhibitor (acetylsalicylic acid), and the venodilator effect was measured. Furthermore, N-desethylamiodarone was infused after oral treatment with hydrocortisone or coinfused with alpha tocopherol. Additional experiments were carried out in bovine aortic endothelial cells to explore the effects of N-desethylamiodarone on the intracellular Ca2+ concentration ([Ca2+]i). RESULTS: N-Desethylamiodarone produced dose-dependent venodilation (47% +/- 4% maximum). In vitro, 10 micromol/L N-desethylamiodarone caused a sustained increase of the endothelial [Ca2+]i. Pretreatment of the volunteers with acetylsalicylic acid reduced the maximum N-desethylamiodarone induced venodilation to 22% +/- 8%; L-NMMA reduced the maximum N desethylamiodarone-induced venodilation to 18% +/- 11%. Pretreatment with acetylsalicylic acid and coinfusion of N-desethylamiodarone and L-NMMA abolished the venodilation, whereas hydrocortisone had no effect. Coinfusion of alpha tocopherol and N-desethylamiodarone reduced the maximum N-desethylamiodarone induced venodilation to 11% +/- 4%. CONCLUSIONS: In concentrations estimated to be in the therapeutic range, N-desethylamiodarone dilates preconstricted human hand veins in vivo and increases endothelial [Ca2+]i in vitro. Subsequently the cyclooxygenase (COX-1) and the endothelial nitric oxide synthase pathways are activated. The resulting venodilation does not involve inflammatory cytokines, inducible nitric oxide synthase, or inducible cyclooxygenase (COX-2). PMID- 10668852 TI - Overconsumption detected by electronic drug monitoring requires subtle interpretation. AB - BACKGROUND: Electronic compliance monitoring has provided new variables to describe drug intake behavior and new strategies to improve compliance. However, as evaluated in this study, the recording of opening events of pill bottles does not necessarily mean drug intake. METHODS: In an open 3-week trial with an oral vitamin combination, drug intake was recorded with use of an electronic pill box that contained 25 capsules and that registered each opening of the bottle. Thirty seven patients were asked to take one capsule every morning for 21 days. Opening and closing events were related to the results of pill counts and patient interviews at the end of the trial. RESULTS: Drug consumption was 101.8% (663 recorded opening and closing events) in the 31 patients who completed the trial. Pill boxes were opened more than once by 10 patients on at least one monitored day. For seven patients the total number of openings was >25 (range, 26 to 29) and thus exceeded the number of capsules provided. A third interview of these patients revealed real overconsumption in only two patients. Six patients remembered that they had shown the device to relatives or friends or that they had checked to see whether they had closed the pill box well, thus turning a "curiosity event" into a drug intake event. CONCLUSION: In short-term studies particularly, such curiosity events may substantially modify the electronic assessment of compliance surrogates. In these trials the combined evaluation of electronic openings, pill counts, and interviews may be a suitable way to reveal such openings without pill intake. PMID- 10668851 TI - In vivo effects of interleukin-10 on human cytochrome P450 activity. AB - BACKGROUND: Injection of lipopolysaccharide into human volunteers leads to an increase in serum interleukin-1beta, interleukin-6, and tumor necrosis factor alpha and a significant decrease in cytochrome P450 (CYP)-mediated drug metabolism. The in vivo effects of the noninflammatory cytokine interleukin-10 (IL-10) on CYP-mediated drug metabolism was examined. METHODS: IL-10 (8 microg/kg) and placebo were administered for 6 days to 12 healthy volunteers in a double-blind crossover study. Tolbutamide (CYP2C9), caffeine (CYP1A2), dextromethorphan (CYP2D6 and CYP3A), and midazolam (CYP3A) were administered on days 4 and 5 to determine individual CYP activities. RESULTS: Few clinically apparent side effects were observed after administration of IL-10; however, blood chemistries reflected an acute-phase response. A significant drop in serum albumin (mean percentage change +/- SD between groups; 4.7% +/- 6.0%, P < or = .02), a significant increase in serum ferritin (736% +/- 717%, P < or = .001), and a significant reduction in platelet count (49% +/- 12%, P < or = .0001) was observed after administration of IL-10. IL-10 significantly (P < or = .02) decreased CYP3A activity 12% +/- 17%, as reflected by midazolam clearance. CYP2C9 activity was significantly (P < or = .005) increased by 38% +/- 35%, as reflected by the tolbutamide urinary metabolic ratio and oral clearance. However, administration of IL-10 resulted in a 40% increase in the fraction unbound of tolbutamide. Therefore no difference in the unbound clearance of tolbutamide was observed between placebo (23.3 +/- 9.7 L/h) or IL-10 (23.5 +/- 11.4 L/h) administration. No significant changes in either CYP1A2 or CYP2D6 activities were observed between placebo and treatment arms of the study. CONCLUSION: IL-10 administration resulted in an acute-phase response. Administration of IL-10 did not alter CYP1A2, CYP2C9, and CYP2D6 activities. CYP3A-mediated biotransformation was reduced by administration of IL-10. PMID- 10668853 TI - CYP3A4 allelic variants with amino acid substitutions in exons 7 and 12: evidence for an allelic variant with altered catalytic activity. AB - OBJECTIVE: To determine the existence of mutant and variant CgammaP3A4 alleles in three racial groups and to assess functions of the variant alleles by complementary deoxyribonucleic acid (cDNA) expression. METHODS: A bacterial artificial chromosome that contains the complete CgammaP3A4 gene was isolated and the exons and surrounding introns were directly sequenced to develop primers to polymerase chain reaction (PCR) amplify and sequence the gene from lymphocyte DNA. DNA samples from Chinese, black, and white subjects were screened. Mutating the affected amino acid in the wild-type cDNA and expressing the variant enzyme with use of the baculovirus system was used to functionally evaluate the variant allele having a missense mutation. RESULTS: To investigate the existence of mutant and variant CgammaP3A4 alleles in humans, all 13 exons and the 5'-flanking region of the human CgammaP3A4 gene in three racial groups were sequenced and four alleles were identified. An A-->G point mutation in the 5'-flanking region of the human CgammaP3A4 gene, designated CgammaP3A4*1B, was found in the three different racial groups. The frequency of this allele in a white population was 4.2%, whereas it was 66.7% in black subjects. The CgammaP3A4*1B allele was not found in Chinese subjects. A second variant allele, designated CgammaP3A4*2, having a Ser222Pro change, was found at a frequency of 2.7% in the white population and was absent in the black subjects and Chinese subjects analyzed. Baculovirus-directed cDNA expression revealed that the CYP3A4*2 P450 had a lower intrinsic clearance for the CYP3A4 substrate nifedipine compared with the wild type enzyme but was not significantly different from the wild-type enzyme for testosterone 6beta-hydroxylation. Another rare allele, designated CgammaP3A4*3, was found in a single Chinese subject who had a Met445Thr change in the conserved heme-binding region of the P450. CONCLUSIONS: These are the first examples of potential function polymorphisms resulting from missense mutations in the CgammaP3A4 gene. The CgammaP3A4*2 allele was found to encode a P450 with substrate-dependent altered kinetics compared with the wild-type P450. PMID- 10668854 TI - Deficient cotinine formation from nicotine is attributed to the whole deletion of the CYP2A6 gene in humans. AB - Nicotine is mainly metabolized to cotinine by cytochrome P450 (CYP) 2A6. Large interindividual differences in nicotine metabolism have been reported in humans. The purpose of this study was to clarify the relationship between the poor metabolism of nicotine and the existence of the CgammaP2A6v1 and CgammaP2A6v2 alleles, and a whole deletion allele of the CgammaP2A6 gene. The plasma concentrations of nicotine and cotinine were measured in 10 healthy subjects after each smoked one cigarette or chewed one piece of nicotine gum. One subject showed no detectable cotinine level in plasma when smoking and the lowest cotinine level when receiving nicotine gum. The subject was regarded as a poor metabolizer of nicotine by a probit analysis and was found to carry a homozygous whole deletion allele of the CgammaP2A6 gene. This is the first report to show that deficient cotinine formation in humans is attributed to the whole deletion of the CgammaP2A6 gene. PMID- 10668855 TI - Antipyretic efficacy of aspirin or acetaminophen. PMID- 10668856 TI - Oxaliplatin: pharmacokinetics and chronopharmacological aspects. AB - Oxaliplatin is the first clinically available diaminocyclohexane platinum coordination complex. The drug is non-cross-resistant with cisplatin or carboplatin and is one of the few active drugs against human colorectal cancer. Its cytotoxicity is synergistic with fluorouracil and folinic acid (leucovorin), the reference treatment for this disease. The main cumulative dose-limiting toxicity of oxaliplatin is peripheral sensory neuropathy. The drug can also produce diarrhoea, vomiting and haematological suppression. Unlike cisplatin, no renal failure or peripheral motor neuropathy have been reported and the sensory neuropathy is partly reversible. Unlike carboplatin, oxaliplatin produces only mild to moderate haematological toxicity. Oxaliplatin undergoes biotransformation into aquated forms in the blood, where 3 species can be found: total platinum, ultrafilterable or 'free' platinum and erythrocyte platinum. Flameless atomic absorption (FAAS) is used for assaying platinum concentration in various tissues. Inductively-coupled plasma mass spectrometry (ICP-MS), with a >10-fold lower sensitivity threshold than FAAS, was also used for the determination of oxaliplatin pharmacokinetics. The pharmacokinetics of oxaliplatin are described by a 3-compartment model. The drug rapidly crosses the cellular membrane as a result of its lipophilicity. Hence, at the end of a 2-hour infusion, approximately 40% of the blood platinum is found in erythrocytes. The distribution half-life of ultrafiltrated plasma platinum ranges from 10 to 25 minutes and its terminal elimination half-life is 26 hours (determined with FAAS) or 270 hours (ICP-MS). The elimination half-life of erythrocytic platinum is 12 to 50 days, close to that of erythrocytes. 30 to 50% of the platinum is recovered in the urine within 2 to 5 days, with renal clearance accounting for half of the total clearance of ultrafiltrated platinum. The total clearance of this species is correlated with the glomerular filtration rate. No pharmacokinetic pharmacodynamic relationship has been established for oxaliplatin. Pharmacokinetic alterations produced by fluorouracil + folinic acid or irinotecan were minimal if any. The prolonged stability of oxaliplatin makes it suitable for continuous infusions over 4 to 5 days, with a delivery rate which can be either constant or chronomodulated (peak rate at 1600h), using programmable ambulatory pumps. Chronomodulation significantly reduces toxicity and improves antitumour activity as compared with constant rate infusion. These differences in pharmacodynamic properties were paralleled by differences in plasma concentration time courses. The different drug concentration profiles achieved with different infusional modalities may be useful tools for understanding the relationship between the pharmacokinetics and pharmacodynamics of oxaliplatin and may lead to further optimisation of its administration schedule and its combination with other drugs. PMID- 10668857 TI - Osmotherapy for elevated intracranial pressure: a critical reappraisal. AB - The administration of osmotic agents is one of the principal strategies to lower elevated intracranial pressure (ICP) and to increase cerebral perfusion pressure. Of the 3 osmotic agents frequently used (mannitol, glycerol and sorbitol), each has characteristic advantages and disadvantages. In addition to renal filtration, sorbitol [elimination half-life (t1/2beta) approximately 1h] and glycerol (t1/2beta 0.2 to 1h) are metabolised, mainly by the liver. The risk of these compounds accumulating in patients with renal insufficiency is low. However, both compounds frequently affect glucose metabolism, leading to an increase in the serum glucose concentration. Mannitol is almost exclusively renally filtered and possesses the slowest elimination from serum (t1/2beta 2 to 4h). The t1/2beta of mannitol is markedly increased in patients with renal insufficiency, but it does not interfere with glucose metabolism. Entry into the cerebrospinal fluid (CSF) is highest with glycerol [CSF: serum ratio of the areas under the concentration time curves (AUC(CSF): AUCs) approximately 0.25], intermediate with mannitol (AUC(CSF): AUCs approximately 0.15) and lowest with sorbitol (AUC(CSF): AUCs approximately 0.10). The elimination of all osmotic agents from the CSF compartment is substantially slower than from serum. During the elimination phase, the CSF-to-serum osmotic gradient is temporarily reversed. This is one cause of the paradoxical rise of ICP above the pretreatment level sometimes observed with osmotherapeutics. The ability of mannitol, glycerol and sorbitol to lower elevated ICP has been extensively documented. However, whether the use of osmotic agents, particularly with repeated application, improves outcome remains unproven. Therefore, these agents should only be used to treat manifest elevations of ICP, not for prophylaxis of brain oedema. PMID- 10668860 TI - Sequence variation of ribosomal internal transcribed spacers (ITS) in commercially important Phytoseiidae mites. AB - Preliminary work is needed to assess the usefulness of different markers at different taxonomic scales when a new group is analyzed, such as the commercially important Phytoseiidae mites. We investigate here the level of sequence variation of the nuclear ribosomal spacers ITS 1 and 2 and the 5.8S gene in six species of Phytoseiidae: Neoseiulus culifornicus, N. fallacis, Euseius concordis, Metaseiulus occidentalis, Typhlodromus pyri and Phytoseiulus persimilis. As expected, the 5.8S gene (148 base pairs) is markedly conserved and displays little variation in between genera comparisons. ITS1 and ITS2 show contrasting patterns: while the ITS2 is short (80-89 bp) and shows little variation, the ITS1 is longer (303-404 bp) and is very variable in sequence. This fact compromises reliable nucleotide homologies when comparing the genera. The comparison of ITS1 sequence similarity at the species level might be useful for species identification, however, the value of ITS in taxonomic studies does not extend to the level of the family. The intraspecific variations of ITS were investigated in three species: N. californicus, N. fallacis and E. concordis. The first species has identical ITS1 sequences and the last two display low polymorphism (2 nucleotide substitutions). The ITS2 and 5.8S sequences were identical in all three subspecies comparisons. PMID- 10668858 TI - Pharmacokinetic-pharmacodynamic consequences and clinical relevance of cytochrome P450 3A4 inhibition. AB - Drug interactions occur when the efficacy or toxicity of a medication is changed by administration of another substance. Pharmacokinetic interactions often occur as a result of a change in drug metabolism. Cytochrome P450 (CYP) 3A4 oxidises a broad spectrum of drugs by a number of metabolic processes. The location of CYP3A4 in the small bowel and liver permits an effect on both presystemic and systemic drug disposition. Some interactions with CYP3A4 inhibitors may also involve inhibition of P-glycoprotein. Clinically important CYP3A4 inhibitors include itraconazole, ketoconazole, clarithromycin, erythromycin, nefazodone, ritonavir and grapefruit juice. Torsades de pointes, a life-threatening ventricular arrhythmia associated with QT prolongation, can occur when these inhibitors are coadministered with terfenadine, astemizole, cisapride or pimozide. Rhabdomyolysis has been associated with the coadministration of some 3 hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors ('statins') and CYP3A4 inhibitors. Symptomatic hypotension may occur when CYP3A4 inhibitors are given with some dihydropyridine calcium antagonists, as well with the phosphodiesterase inhibitor sildenafil. Excessive sedation can result from concomitant administration of benzodiazepine (midazolam, triazolam, alprazolam or diazepam) or nonbenzodiazepine (zopiclone and buspirone) hypnosedatives with CYP3A4 inhibitors. Ataxia can occur with carbamazepine, and ergotism with ergotamine, following the addition of a CYP3A4 inhibitor. Beneficial drug interactions can occur. Administration of a CYP3A4 inhibitor with cyclosporin may allow reduction of the dosage and cost of the immunosuppressant. Certain HIV protease inhibitors, e.g. saquinavir, have low oral bioavailability that can be profoundly increased by the addition of ritonavir. The clinical importance of any drug interaction depends on factors that are drug-, patient- and administration-related. Generally, a doubling or more in plasma drug concentration has the potential for enhanced adverse or beneficial drug response. Less pronounced pharmacokinetic interactions may still be clinically important for drugs with a steep concentration-response relationship or narrow therapeutic index. In most cases, the extent of drug interaction varies markedly among individuals; this is likely to be dependent on interindividual differences in CYP3A4 tissue content, pre existing medical conditions and, possibly, age. Interactions may occur under single dose conditions or only at steady state. The pharmacodynamic consequences may or may not closely follow pharmacokinetic changes. Drug interactions may be most apparent when patients are stabilised on the affected drug and the CYP3A4 inhibitor is then added to the regimen. Temporal relationships between the administration of the drug and CYP3A4 inhibitor may be important in determining the extent of the interaction. PMID- 10668859 TI - Clinical pharmacokinetics of transdermal opioids: focus on transdermal fentanyl. AB - Transdermal delivery allows continuous systemic application of opioids through the intact skin. This review analyses the pharmacokinetic properties of transdermal opioid administration in the context of clinical experience, with a focus on fentanyl. A transdermal therapeutic system (TTS) for fentanyl has been developed. The amount of fentanyl released is proportional to the surface area of the TTS, which is available in different sizes. After the first application of a TTS, a fentanyl depot concentrates in the upper skin layers and it takes several hours until clinical effects are observed. The time from application to minimal effective and maximum serum concentrations is 1.2 to 40 hours and 12 to 48 hours, respectively. Steady state is reached on the third day, and can be maintained as long as patches are renewed. Within each 72-hour period, serum concentrations decrease gradually over the second and third days. When a TTS is removed, fentanyl continues to be absorbed into the systemic circulation from the cutaneous depot. The terminal half-life for TTS fentanyl is approximately 13 to 25 hours. The interindividual variability of serum concentrations, partly caused by different clearance rates, is markedly larger than the intraindividual variability. The effectiveness of TTS fentanyl was first demonstrated in acute postoperative pain. However, the slow pharmacokinetics and large variability of TTS fentanyl, together with the relatively short duration of postoperative pain, precluded adequate dose finding and led to inadequate pain relief or, especially, a high incidence of respiratory depression; such use is now contraindicated. Conversely, in cancer pain, TTS fentanyl offers an interesting alternative to oral morphine, and its effectiveness and tolerability in this indication has been demonstrated by a number of trials. Its usefulness in chronic pain of nonmalignant origin remains to be confirmed in controlled trials. In general, TTS fentanyl produces the same adverse effects as other opioids, mainly sedation, nausea, vomiting and constipation. In comparison with oral morphine, TTS fentanyl causes fewer gastrointestinal adverse events. The risk of hypoventilation is comparatively low in cancer patients. Sufentanil and buprenorphine may also be suitable for transdermal delivery, but clinical results are not yet available. Transdermal morphine is only useful if applied to de-epithelialised skin. However, iontophoresis may allow transdermal administration of opioids, including morphine, with a rapid achievement of steady state concentrations and the ability to adjust delivery rates. This would be beneficial for acute and/or breakthrough pain, and initial clinical trials are in progress. PMID- 10668861 TI - Chorioptic mange (Acarina: Psoroptidae) in domestic and wild ruminants in Israel. AB - In a 20-year-survey, 9364 dairy cattle in 324 herds kept under a zero-grazing management, 1252 beef cattle in 46 herds grazing all the year round, 3347 sheep in 134 herds (only 26 are grazing herds), and 872 goats in 47 herds (only 20 are grazing) were examined. The mites collected from cattle were identified as Chorioptes texanus only, and those from sheep, goats and gazelles were identified as C. bovis. Chorioptic mange was not diagnozed in grazing beef cattle, ibexes and housed animals (as compared to grazing herds), and in rams and billy goats. Holstein-Israeli bulls kept in insemination centres were not clinically infested, whereas four Charolais bulls were infested with chorioptic mange. Infestation rate was higher in older animals than in younger ones. Hoggets and young goats over 10 months and heifers over 13 months were found clinically infested with Choriopic mites. Lesions were not usually extensive and occurred mainly in predilection sites. The ocular form in sheep and the groin form in goats are very uncommon and apparently are reported for the first time. Chorioptic mange was recorded generally throughout the year. In our study seasonal distribution could not be demonstrated in cattle, while in sheep and goats the highest infestation rate occurred in February to March and the lowest in August and September. PMID- 10668862 TI - High temperatures eliminate Wolbachia, a cytoplasmic incompatibility inducing endosymbiont, from the two-spotted spider mite. AB - Wolbachia can induce cytoplasmic incompatibility (CI) in the arrhenotokous two spotted spider mite between uninfected females and infected males. Cytoplasmic incompatibility is expressed through a male-biased sex ratio and a low hatchability, and can be suppressed by removing Wolbachia from spider mites reared on a diet with antibiotics. Here we investigated whether heat-treatment can elimate Wolbachia from infected mites. Using a PCR assay with a Wolbachia specific primer pair (ftsZ), and by standard crosses, we were able to show that 71 per cent of the mites had lost the Wolbachia infection after rearing the infected population at 32+/-0.5 degrees C for four generations. The infection could be completely removed when mites were reared at 32+/-0.5 degrees C for six generations. Curing through high temperatures could be one of the reasons why mixed infected/uninfected populations occur in the field. An additional consequence of rearing mites at 32+/-0.5 degrees C was the shortened development time. The effect of environmental temperature on the abundance of Wolbachia and possible behavioural consequences for the spider mite are discussed. PMID- 10668863 TI - Sequence variations in the Boophilus microplus Bm86 locus and implications for immunoprotection in cattle vaccinated with this antigen. AB - Cattle tick infestations constitute a major problem for the cattle industry in tropical and subtropical regions of the world. Traditional control methods have been only partially successful, hampered by the selection of chemical-resistant tick populations. The Boophilus microplus Bm86 protein was isolated from tick gut epithelial cells and shown to induce a protective response against tick infestations in vaccinated cattle. Vaccine preparations including the recombinant Bm86 are used to control cattle tick infestations in the field as an alternative measure to reduce the losses produced by this ectoparasite. The principle for the immunological control of tick infestations relies on a polyclonal antibody response against the target antigen and, therefore, should be difficult to select for tick-resistant populations. However, sequence variations in the Bm86 locus, among other factors, could affect the effectiveness of Bm86-containing vaccines. In the present study we have addressed this issue, employing data obtained with B. microplus strains from Australia, Mexico, Cuba, Argentina and Venezuela. The results showed a tendency in the inverse correlation between the efficacy of the vaccination with Bm86 and the sequence variations in the Bm86 locus (R2 = 0.7). The mutation fixation index in the Bm86 locus was calculated and shown to be between 0.02 and 0.1 amino acids per year. Possible implications of these findings for the immunoprotection of cattle against tick infestations employing the Bm86 antigen are discussed. PMID- 10668864 TI - Monooxygenases play only a minor role in resistance to synthetic pyrethroids in the cattle tick, Boophilus microplus. AB - We investigated the role of monooxygenases in resistance to synthetic pyrethroids (SPs) in the cattle tick, Boophilus microplus. We found that monooxygenases play only a minor role in resistance to SPs in both resistant and susceptible strains of B. microplus. We blocked the monooxygenases with piperonyl butoxide (PBO) and simultaneously applied the SPs, flumethrin and cypermethrin to larval B. microplus. PBO increased the effect of flumethrin (synergism ratios 2.7-8.9) more than it increased the effect of cypermethrin (synergism ratios 1.9-3.1). Of the four strains tested, Parkhurst, which is resistant to SPs, was the least affected by the addition of PBO (synergism ratios after cypermethrin was applied 1.9; after flumethrin 2.7) whereas N.R.F.S., the strain susceptible to SPs, was the most affected by synergism between PBO and SPs (synergism ratio after cypermethrin was applied 3.1; after flumethrin 8.9). We hypothesize that B. microplus lacks monooxygenases capable of conferring resistance to SPs because it and its recent ancestors were blood-feeders rather than herbivores. PMID- 10668865 TI - Detecting resistance to organophosphates and carbamates in the cattle tick Boophilus microplus, with a propoxur-based biochemical test. AB - Rapid and sensitive detection of resistance to insecticides in arthropods is needed. In the cattle tick. Boophilus microplus, resistance to a variety of acaricides is widespread. The most commonly used assay for resistance, the larval packet test, takes at least two, but generally six weeks for a one-host tick like B. microplus to complete and may take up to three months to complete for three host ticks. Here we describe a test for resistance to organophosphate acaricides that can be used on larvae and adult ticks which takes less than 24 hours. The test measures the difference in acetylcholinesterase (AChE) activity in homogenates of ticks in the presence and absence of propoxur, a carbamate acaricide. We found clear discrimination of organophosphate-susceptible and organophosphate-resistant adults with 100 microM propoxur. AChE from susceptible ticks had almost no activity at this concentration of propoxur whereas AChE from resistant ticks had 67% of its potential activity. AChE from heterozygote ticks could also be distinguished from AChE from homozygous-susceptible and homozygous resistant ticks. This is the first biochemical test for resistance to an acaricide. Rapid, sensitive tests like ours will allow resistance to organophosphates to be detected soon after it develops in the field, thus, the spread of resistance might be slowed and the useful life of acaricides extended. PMID- 10668866 TI - Sequential histopathology at the Rhipicephalus sanguineus tick feeding site on dogs and guinea pigs. AB - The tick Rhipicephalus sanguineus is a very common parasite of dogs worldwide. Dogs seem unable to acquire resistance against this tick species, whereas guinea pigs demonstrate a very strong resistance following primary infestation. We studied the inflammatory reaction at the R. sanguineus tick feeding site on dogs and guinea pigs during primary and tertiary infestations at different time intervals after attachment. Biopsies were collected after 4, 24, 48 and 96 hours. Changes that were found in all experimental groups included a cone of cement around the mouthparts of the tick, epidermal hyperplasia, edema and inflammatory cell infiltration in the dermis directly underneath the tick attachment site. Dogs reacted to ticks mainly with neutrophils, particularly after repeated exposure. Mast cells and mononuclear leukocytes were also present. Guinea pigs reacted to R. sanguineus mainly with mononuclear cells, eosinophils and basophils. These cells were particularly numerous after repeated exposure to R. sanguineus. Our results suggest that basophils and eosinophils are involved in resistance of guinea pigs to R. sanguineus and that neutrophils in dogs have little effect against this tick species. PMID- 10668867 TI - Rural surgery: opportunity or minefield. PMID- 10668868 TI - Advances in the management of acute respiratory distress syndrome: protective ventilation. AB - The approach to mechanical ventilation has been revolutionized by new insights into the pathogenesis of respiratory failure in acute respiratory distress syndrome (ARDS). Concepts such as low-volume ventilation, permissive hypercapnia, inverse ratio ventilation, best and intrinsic positive end-expiratory pressure, airway shear, pressure volume curves, inflection points, and prone positioning have radically transformed thinking about ventilator management. Since 1966, more than 8000 ARDS-related publications have appeared. Studies highlighting the experimental basis for innovations in mechanical ventilation are presented. Selected clinical series that exemplify the use of these new strategies are reviewed, to demonstrate how key experimental and clinical research has altered our understanding about what works, and why. Mismanagement of mechanical ventilation causes lung injury and increases mortality. The strategy of protective ventilation has provided the first substantial reduction of mortality in the history of ARDS. PMID- 10668869 TI - Internal drainage of giant acute pseudocysts: the role of video-assisted pancreatic necrosectomy. AB - BACKGROUND: Internal drainage of giant pancreatic pseudocysts secondary to acute pancreatitis is frequently complicated with postoperative retroperitoneal infection and hemorrhage. Recent data suggest that the risk factor is unrecognized pancreatic necrosis; presumably, pancreatic necrosis becomes infected with bacteria introduced by the cystoenteric anastomosis. HYPOTHESIS: Video-assisted pancreatic necrosectomy, performed at the time of internal drainage, may prevent postoperative retroperitoneal complications in patients with giant acute pseudocysts. DESIGN: A consecutive case-series. SETTING: An urban, university-affiliated, tertiary referral center. PATIENTS: Ten consecutive patients with acute pseudocysts measuring 10 cm or more in major diameter. The mean extent of pancreatic necrosis, as shown by contrast-enhanced computed tomography, was 50%. All patients were operated on electively, at an average time of 7.7 weeks from onset of the attack to surgical treatment. INTERVENTION: Through a midline incision, a 4-cm opening is made at the base of the pseudocyst. Standard laparoscopic instruments are introduced into the pseudocyst and video assisted pancreatic necrosectomy is performed. The opening is then anastomosed to a Roux-en-Y limb of the jejunum. MAIN OUTCOME MEASURES: Feasibility and safety of video-assisted pancreatic necrosectomy, postoperative morbidity and mortality, hospital stay, and resolution of pseudocysts. RESULTS: Complete necrosectomy was safely performed throughout. There were neither postoperative retroperitoneal complications nor mortality. Mean hospital stay was 8.2 days and all pseudocysts resolved at a mean follow-up of 6.9 months. CONCLUSIONS: Video-assisted pancreatic necrosectomy at the time of internal drainage seems to prevent postoperative retroperitoneal complications in patients with giant acute pseudocysts. Depending on appropriate surgical timing, video-assisted necrosectomy is a feasible and safe procedure. PMID- 10668870 TI - Risk factors for postoperative hypocalcemia after surgery for primary hyperparathyroidism. AB - HYPOTHESIS: A variety of clinical and biochemical variables may be associated with hypocalcemia after surgery for parathyroid adenoma. DESIGN: A prospective study of patients who underwent surgery for solitary parathyroid adenoma. SETTING: A university hospital department of surgery. PATIENTS: Eighty-six consecutive patients who underwent surgery for solitary parathyroid adenoma. INTERVENTION: Parathyroidectomy according to the principles of unilateral neck exploration. MAIN OUTCOME MEASURES: Clinical and biochemical risk factors for early (< or =4 days after surgery) and late (1 year after surgery) postoperative symptomatic and biochemical hypocalcemia. RESULTS: Twenty-two patients developed early symptomatic hypocalcemia. The difference in total serum calcium levels between patients, with and without early symptomatic hypocalcemia, was evident on the third and fourth postoperative days. Serum level of osteocalcin greater than 6.0 microg/L, bilateral neck exploration, and history of cardiovascular disease were risk factors for symptomatic hypocalcemia (odds ratios [95% confidence intervals]: 4.4 [1.4-14.1], 3.8 [1.3-11.6], and 0.1 [0.02-0.60], respectively). Patients with up to 1 risk factor had a possibility of only 7% to develop early symptomatic hypocalcemia. One year after surgery, 16 patients had low levels of total serum calcium (late biochemical hypocalcemia) and were asymptomatic. Preoperative intermittent hypercalcemia was associated with an increased risk for late biochemical hypocalcemia (odds ratio, 3.9; 95% confidence interval, 1.0 16.3). CONCLUSIONS: Clinical and biochemical risk factors for early and late postoperative hypocalcemia in patients who underwent surgery for solitary parathyroid adenoma were found. A clinically useful prognostic index for early symptomatic hypocalcemia was constructed using these risk factors. PMID- 10668871 TI - Evaluation of magnetic resonance cholangiography in the management of bile duct stones. AB - HYPOTHESIS: Magnetic resonance cholangiography (MRC) offers the potential for accurate, noninvasive detection of common bile duct stones (CBDSs) before cholecystectomy, and for a consequent reduction in the incidence of preoperative negative diagnoses associated with endoscopic retrograde cholangiography (ERC). DESIGN: Prospective cohort study: MRC results were correlated with ERC (high-risk patients) or intraoperative cholangiography (moderate-risk patients). SETTING: A university hospital providing primary, secondary, and tertiary care. PATIENTS: Seventy patients with suspected CBDSs scheduled to undergo elective cholecystectomy between April 15, 1997, and September 30, 1998. Forty patients were considered at high risk and 30 at moderate risk for CBDSs, according to results of liver function tests and sonograms of the upper abdomen. MAIN OUTCOME MEASURES: Confirmation or exclusion of CBDSs by MRC was assessed by a panel of radiologists who were unaware of the ERC results. Results of ERC and intraoperative cholangiography were analyzed by the investigating gastroenterologists or surgeon. RESULTS: Results of MRC were positive for CBDSs in 21 (52%) of 40 high-risk patients, a finding confirmed by preoperative ERC in 19 (90%) of 21 patients. Results of MRC were positive for CBDSs in 6 (20%) of 30 moderate-risk patients, all of which were confirmed by intraoperative cholangiography. Finally, CBDSs were present in 19 (48%) of 40 high-risk patients and 6 (20%) of 30 moderate-risk patients (P = .02). Overall sensitivity and specificity of MRC were 100% and 95.6%, respectively; the positive and negative predictive values were 92.6% and 100%, respectively. CONCLUSIONS: Magnetic resonance cholangiography is a reliable, noninvasive method for the detection or exclusion of CBDSs, and seems to reduce the frequency of negative diagnoses associated with ERC. Magnetic resonance cholangiography revealed no CBDSs in 19 (48%) of 40 patients at high risk for CBDSs. Thus, MRC-based diagnosis has the potential to reduce the number of invasive preoperative diagnostic procedures and their associated risks and overall health care costs. PMID- 10668872 TI - Management of benign biliary strictures: biliary enteric anastomosis vs endoscopic stenting. AB - HYPOTHESIS: Although advances in endoscopic procedures have provided alternative options for relieving biliary obstructions, the overall chance of cure for patients with benign biliary stricture is the same using surgical or endoscopic treatment. DESIGN: Case-control study. SETTING: Tertiary care university hospital. PATIENTS: Of 163 patients referred for treatment with diagnoses of benign strictures of the common bile duct between January 1, 1975, and July 1, 1998, we studied 42 patients with postcholecystectomy stricture and a follow-up longer than 60 months. Twenty of these patients were treated with endoscopic stenting and 22 with surgery (hepaticojejunostomy, choledochojejunostomy, or intrahepatic cholangiojejunostomy). MAIN OUTCOME MEASURES: Postoperative mortality and morbidity and long-term outcome. The rate of restenosis was also determined. RESULTS: Morbidity occurred more frequently in patients treated with endoscopic procedures than with surgical ones (9 vs 2; P = .34). Hospital mortality was 0%. Surgery achieved excellent or good long-term outcome in 17 of 22 patients. Endoscopic biliary stenting was successful in 16 of 20 patients. Overall, excellent or good outcomes were achieved in 34 patients (81%). CONCLUSION: The ability to achieve steady, long-term results confirms hepaticojejunostomy as the best procedure in the treatment of benign biliary strictures, even if endoscopic procedures are gaining a new role in the treatment of a greater number of patients. PMID- 10668873 TI - Ultrasound can estimate the pathologic size of infiltrating ductal carcinoma. AB - HYPOTHESIS: Ultrasound (US) of the breast will accurately measure breast tumor size when compared with size as determined by pathologic examination. DESIGN: Retrospective case series. SETTING: University hospital-based breast center. PATIENTS: Thirty-five women with a diagnosis of breast cancer who had US as a component of their evaluation. MAIN OUTCOME MEASURE: Tumor size as measured by US compared with size measured by pathologic examination. RESULTS: Size measured by US ranged from 0.45 to 3.81 cm. Size measured by pathologic examination ranged from 0.5 to 5 cm. The mean difference of size measured by US vs pathologic size was 0.4 cm (P = .01). When only tumors with invasive ductal histology are evaluated, the mean difference in size is 0.33 cm (P = .008). The range of difference was -1.6 cm to +0.42 cm. Seventeen percent of invasive ductal tumors were underestimated by more than 1 cm; none were underestimated by more than 2 cm. CONCLUSIONS: This study demonstrates that, although US tends to underestimate the pathologic tumor size, 83% of invasive ductal tumors fall within a 1-cm and 100% fall within a 2-cm extension of the US-measured tumor size. Therefore, it is possible to use US to monitor the extent of treatment size when developing very localized therapeutic tools. PMID- 10668874 TI - Effects of fluconazole administration in critically ill patients: analysis of bacterial and fungal resistance. AB - HYPOTHESIS: The administration of fluconazole in intensive care unit (ICU) patients leads to the emergence of bacterial and fungal resistance. DESIGN: Retrospective analysis of 2 patient cohorts: (1) critically ill patients treated in surgical, trauma, and medical ICUs between June 1997 and January 1999 who did and did not receive fluconazole; and (2) ICU patients with fungal infections and sensitivity testing results from June 1994 to December 1998. SETTING: University affiliated tertiary care hospital. PATIENTS: The first cohort included 99 ICU patients with documented microorganism culture(s) who were treated with (n = 50) or without (n = 49) fluconazole; the second cohort included 38 patients with Candida species infection, identification, and antifungal susceptibility testing. RESULTS: Mortality (40% vs 20%; P = .03) and hospital length of stay (33.8 vs 25.6 days; P = .04) were higher in the patients treated with fluconazole compared with patients not treated with fluconazole. The ICU length of stay was also higher in patients treated with fluconazole (23.7 vs 15.1 days; P = .009). An increase in bacterial resistance occurred in patients after fluconazole treatment as opposed to bacterial resistance of patients who were treated for bacterial microorganism(s) without fluconazole (16% vs 4%; P = .049). Comparison of patient populations with Candida species identification before and after December 1997 showed an increase in Candida species resistance to fluconazole (11% vs 36%; P = .16), respectively. Fungal strains were dominated by a combination of Candida albicans and Candida glabrata in both populations (60% [before 1998] vs 82% [after 1998]), with an emergence of Candida non-albicans species tolerant to fluconazole. The amount of fluconazole administered and the number of patients receiving fluconazole treatment in the ICUs has also increased when comparing both periods. CONCLUSIONS: Comparison of critically ill patient populations with and without fluconazole treatment found increased mortality and longer hospital and ICU lengths of stay in the fluconazole-treated group. This group also had higher bacterial pathogen resistance to antibiotics after fluconazole administration compared with bacterial resistance of patients without fluconazole treatment. Our results warrant concern regarding worsening bacterial infections, increased mortality, and an increase in Candida resistance to fluconazole from increased use in ICU patients, with a shift in yeast infection that is more difficult to treat. PMID- 10668876 TI - Accuracy of ultrasonography in the diagnosis of peritonitis compared with the clinical impression of the surgeon. AB - HYPOTHESIS: Peritonitis is a well-known indication for surgery, but its preoperative cause usually is not established. We hypothesize that abdominal ultrasonography is superior to the clinical impression of the surgeon in detecting the cause of peritonitis. DESIGN: A prospective case series. SETTING: A major university hospital in Taiwan, Republic of China. PATIENTS AND METHODS: One hundred two patients with a diagnosis of peritonitis admitted to the Department of Emergency Medicine, National Taiwan University Hospital, Taipei, were included in this study. All 102 patients underwent an abdominal ultrasonographic examination; and the ultrasonographic findings of these patients were classified into 2 categories: positive findings and normal screening results. The accuracy of clinical impression in detecting the cause of peritonitis was compared with the accuracy of abdominal ultrasonography. RESULTS: Ultrasonography and clinical impression accurately diagnosed the peritonitis in 85 (83.3%) and 52 (51.0%) of the patients, respectively. The difference between ultrasonography and clinical impression in the diagnosis of peritonitis was significant (P<.001). Among 45 patients without a preoperative clinical diagnosis, a diagnosis was made by ultrasonography for 32 (71%) of them. There were a total of 98 patients with positive ultrasonographic findings, and 4 patients had normal screening results. Of the 98 patients with positive ultrasonographic findings undergoing surgery, all had abdominal pathological characteristics. The 4 patients with normal screening results received nonoperative treatment. CONCLUSIONS: Ultrasonography is a more sensitive technique than clinical judgment in diagnosing peritonitis. Ultrasonography may be a useful diagnosing modality in patients with peritonitis in whom the clinical cause is unclear. PMID- 10668875 TI - Repair of chronic anorectal fistulae using commercial fibrin sealant. AB - HYPOTHESIS: Commercially produced fibrin sealant can be used to completely close both simple and complex fistulae in ano. METHODS: A 29-patient prospective nonrandomized clinical trial was performed. In the operating room, the patient underwent an examination with anesthesia and the primary and secondary fistula tract openings were attempted to be identified. The fistula tract was curetted and fibrin sealant was injected into the secondary fistula tract opening until fibrin sealant was seen coming from the primary opening. A petroleum jelly gauze was then applied over the secondary opening and the patient was sent home. Follow up visits were scheduled for 1 week, 1 month, 3 months, and 1 year later. RESULTS: Twenty-nine consecutive patients received fibrin sealant injections for their fistulae in ano, with a mean follow-up of 6 months. Two patients had a history of Crohn disease (regional enteritis) and 2 patients had human immunodeficiency virus infection. Overall, 17 (68%) of 25 patients have had successful closure of their fistula with 4 patients lost to follow-up. Two patients required reinjection with fibrin sealant, and neither of these subsequently had closure. One of the 2 patients with Crohn disease had closure, as well as 1 human immunodeficiency virus-positive patient. In addition, there has been no evidence of incontinence or complications related to the use of fibrin sealant in this procedure. CONCLUSIONS: Initial results in the treatment of chronic anorectal fistulae using commercial fibrin sealant are optimistic, but require further support through longer follow-up data. Fibrin sealant treatment of anorectal fistulae offers a unique mode of management which is safe, simple, and easy for the surgeon to perform. By using fibrin sealant, the patient avoids the risk of fecal incontinence and the discomfort of prolonged wound healing that may be associated with fistulotomy. PMID- 10668877 TI - Restoration of body temperature to normothermia during resuscitation following trauma-hemorrhage improves the depressed cardiovascular and hepatocellular functions. AB - HYPOTHESIS: Rewarming the body to 37 degrees C during resuscitation following trauma-hemorrhage has salutary effects on cardiovascular and hepatocellular functions. DESIGN, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Male rats underwent laparotomy (trauma induced) and were then bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the maximum shed blood volume was returned in the form of Ringer lactate solution. Rats were exposed to ambient temperature and allowed to become hypothermic during hemorrhage. The animals were then resuscitated with 4 times the volume of shed blood with Ringer lactate solution for 60 minutes. In 1 group, the body temperature was rewarmed to 37 degrees C during resuscitation. In another group, the body temperature was maintained at hypothermia (32 degrees C) for 4 hours after resuscitation. In an additional group, the body temperature was kept at 37 degrees C during hemorrhage and resuscitation. At 4 hours after resuscitation, the rats were returned to a room with ambient temperature. Various in vivo heart performance variables (maximal rate of pressure increase and decrease), cardiac output, hepatocellular function, and plasma IL-6 level were determined at 24 hours after resuscitation. RESULTS: Either maintenance of normothermia during hemorrhage or prolonged hypothermia following resuscitation had deleterious effects on cardiovascular variables and hepatocellular function and up-regulated plasma IL-6 levels. In contrast, rewarming the body to 37 degrees C during resuscitation improved cardiac contractility, cardiac output, and hepatocellular function and reduced plasma IL-6 level. CONCLUSION: Since rewarming the body temperature to normothermia during resuscitation improved depressed cardiovascular and hepatocellular functions, this should be considered as a useful adjunct to fluid resuscitation after trauma-hemorrhage. PMID- 10668878 TI - Stage III colon cancers: why adjuvant chemotherapy is not offered to elderly patients. AB - HYPOTHESIS: Adjuvant chemotherapy is not offered to elderly patients with stage III colon cancer. DESIGN: A retrospective review of hospital and office records. SETTING: A suburban community hospital. PATIENTS: The medical records of 69 patients with stage III colon cancer were reviewed. All identified from the Tumor Registry at Jersey Shore Medical Center, Neptune, NJ, were included in this study. RESULTS: Thirty-five patients (51%) did not receive adjuvant chemotherapy. After adjusting for age, women were 5.8 times less likely to receive chemotherapy (P = .002). Patients not receiving chemotherapy were significantly older (78.7 vs 70.4 years; P = .003) than those who received adjuvant chemotherapy. There was no relation found between the year of diagnosis and the administration of chemotherapy. There were 4 major reasons for not receiving chemotherapy: (1) not offered (n = 12, 34%), (2) refused (n = 11, 31%), (3) too old (n = 7, 20%), and (4) significant concomitant disease (n = 5, 14%). CONCLUSIONS: A large group of elderly patients who had been surgically treated for colon cancer and who were eligible for adjuvant chemotherapy either were not referred for treatment or refused treatment. This suggests a bias on the part of surgeons, primary care physicians, and patients against the use of chemotherapy in elderly patients. PMID- 10668879 TI - Long-term results of subcutaneous parathyroid grafts in uremic patients. AB - HYPOTHESIS: Parathyroid glands are normally surrounded (entirely or partially) by fatty tissue. Subcutaneous parathyroid grafts are thus located in a normal environment. Therefore, we postulated that the late results of subcutaneous implantation of parathyroid tissue in uremic patients should be at least as good as those reported for intramuscular grafting. We also challenged the idea that the recurrence rate of renal hyperparathyroidism after surgery depended solely on the type of hyperplasia (diffuse vs nodular) observed in the implanted tissue. DESIGN: A retrospective study of a series of patients without loss to follow-up. SETTING: A university hospital and 9 affiliated dialysis units. PATIENTS AND INTERVENTIONS: Fifty-nine patients (33 women and 26 men) operated on for renal hyperparathyroidism underwent the resection of at least 4 parathyroid glands followed by presternal subcutaneous implantation of parathyroid tissue. They were followed up for 12 to 130 months (median, 38 months). MAIN OUTCOME MEASURES: Failure of treatment, recurrence of disease, and hypoparathyroidism. RESULTS: During the study period, 9 patients had to undergo another operation: 2 (3%) for persistent hyperparathyroidism due to a fifth ectopic gland and 7 (12%) for recurrence of hyperparathyroidism resulting from hypertrophy of the subcutaneous grafts. Four patients received a kidney transplant. The prevalence of hypoparathyroidism (intact parathyroid hormone serum level <1.6 pmol/L with a normal or low serum calcium concentration) was 14% (8 of 59 patients), and the curve representing the distribution of intact parathyroid hormone serum concentrations among operated on patients was shifted to the left when compared with the curve of patients who underwent hemodialysis and who had no indication for parathyroid surgery. In this latter group, the peak of the curve was situated between 1 and 2 times the upper normal limit, while it was in the normal range 12 to 130 months after total parathyroidectomy and subcutaneous parathyroid autotransplantation. No relation was observed between the recurrence rate of the disease and the histological characteristics of the parathyroid grafts. Also, their function was not influenced by the presence or absence of aluminum deposits in bone biopsy specimens that were obtained at the time of cervical exploration. CONCLUSIONS: The late results of total parathyroidectomy and presternal subcutaneous grafting compare favorably with the published data on other surgical techniques proposed for the treatment of renal hyperparathyroidism. The ease with which the hypertrophied grafts are removed when the disease recurs warrants further use of this procedure. PMID- 10668880 TI - Optimizing cardiovascular gene therapy: increased vascular gene transfer with modified adenoviral vectors. AB - BACKGROUND: Adenovirus is widely used as a vector for gene transfer to the vasculature. However, the efficiency of these vectors can be limited by ineffective viral-target cell interactions. Viral attachment, which largely determines adenoviral tropism, is mediated through binding of the adenoviral fiber coat protein to the Coxsackievirus and adenovirus receptor, while internalization follows binding of the adenoviral RGD motif to alpha(v)-integrin receptors. Modifications of the fiber coat protein sequence have been successful for targeting the adenovirus to more prevalent receptors in the vasculature, including heparan sulfate-containing receptors and alpha(v)-integrin receptors. HYPOTHESIS: Modified adenoviral vectors targeted to receptors more prevalent in the vasculature result in an increased transfer efficiency of the virus in vitro and in vivo even in the presence of clinically relevant doses of heparin. DESIGN: We tested 2 modified E1- and E3-deleted Ad5 type adenoviral vectors containing the beta-galactosidase gene. AdZ.F(pK7) contains multiple positively charged lysines in the fiber coat protein that target the adenovirus to heparan sulfate receptors, while AdZ.F(RGD) contains an RGD integrin-binding sequence in the fiber coat protein that allows binding to alpha(v)-integrin receptors. The gene transfer efficiency of these modified viruses was compared in rat aortic smooth muscle cells in vitro and in an in vivo porcine model of balloon-induced arterial injury. Because of the use of heparin during most vascular surgical procedures and the concern that heparin might interfere with the binding of AdZ.F(pK7) to heparan sulfate receptors, the effect of heparin on the in vitro and in vivo transfer efficiency of these 2 modified adenoviruses was evaluated. RESULTS: In vitro infection of rat aortic smooth muscle cells with AdZ.F(pK7) and AdZ.F(RGD) resulted in significantly higher levels of beta-galactosidase expression compared with the unmodified adenovirus (mean +/- SEM, 1766.3 +/- 89.1 and 44.8 +/- 3.4 vs 10.1 +/- 0.7 mU per milligram of protein; P<.001). Following heparin administration, the gene transfer efficiency achieved with AdZ.F(pK7) diminished slightly in a concentration-dependent manner. However, the transfer efficiency was still greater than with the unmodified virus (mean +/- SEM, 1342.3 +/- 101.8 vs 4.8 +/- 0.4 mU per milligram of protein; P<.001). In vivo, following injury to the pig iliac artery with a 4F Fogarty balloon catheter, we found that AdZ.F(pK7) transduced the artery approximately 35-fold more efficiently than AdZ.F and 3 fold more efficiently than AdZ.F(RGD) following the administration of intravenous heparin, 100 U/kg body weight, and heparinized saline irrigation. CONCLUSIONS: Modifications of the adenovirus that lead to receptor targeting resulted in significantly improved gene transfer efficiencies. These improvements in transfer efficiencies observed with the modified vectors decreased slightly in the presence of heparin. However, AdZ.F(pK7) was still superior to AdZ.F(RGD) and AdZ.F despite heparin administration. These data demonstrate that modifications of adenoviral vectors that enhance binding to heparan sulfate receptors significantly improve gene transfer efficiency even in the presence of heparin and suggest an approach to optimize gene transfer into blood vessels. PMID- 10668881 TI - Plasma alpha-glutathione S-transferase: a sensitive indicator of hepatocellular damage during polymicrobial sepsis. AB - HYPOTHESIS: Since studies have found the liver enzyme alpha-glutathione S transferase (alphaGST) to be a marker of hepatic injury after hemorrhagic shock, alphaGST also may serve as a sensitive indicator of hepatocellular damage during the early stage of polymicrobial sepsis. DESIGN, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Male adult rats were subjected to the cecal ligation and puncture (CLP) model of polymicrobial sepsis or sham operation, followed by fluid resuscitation with isotonic sodium chloride solution. Systemic blood samples were taken at 2, 5, 10, or 20 hours after CLP or sham operation. Plasma levels of alphaGST and lactate were determined using an enzyme immunoassay and enzymatic assay, respectively. Additional animals were examined for morphologic alterations in liver ultrastructure of septic animals using electron microscopy. RESULTS: A similar level of alphaGST (mean +/- SEM, 30.5 +/- 3.5 microg/L) was found in the sham group at all measured time points. Although plasma levels of alphaGST did not change at 2 hours after CLP, they were elevated by 249% at 5 hours after the onset of sepsis and continued to increase throughout the septic course. Plasma lactate levels were significantly increased only at 20 hours after CLP (P<.001). Previous studies have shown that liver transaminase levels did not increase at 5 hours, but at 10 and 20 hours after CLP. In addition, electron microscopy revealed structural changes in hepatocyte morphology at 5 and 20 hours after CLP that were indicative of hepatocellular injury. CONCLUSION: Since plasma alphaGST levels increased earlier than plasma lactate and liver transaminase levels, alphaGST may be a more sensitive indicator of early liver injury and should be used in monitoring hepatocellular damage during the progression of sepsis. PMID- 10668882 TI - A prospective evaluation of recurrent laryngeal nerve paralysis during thyroidectomy. AB - HYPOTHESIS: Recurrent laryngeal nerve paralysis after thyroidectomy can be unrecognized without routine laryngoscopy, and patients have a good potential for recovery during follow-up. DESIGN: A prospective evaluation of vocal cord function before and after thyroidectomy. Periodic vocal cord assessment was performed until recovery of cord function. Persistent cord palsy for longer than 12 months after the operation was regarded as permanent. SETTING: A university hospital with about 150 thyroid operations performed by 1 surgical team per year. PATIENTS: From January 1, 1995, to April 30, 1998, 500 consecutive patients (84 males and 416 females) with documented normal cord function at the ipsilateral side of the thyroidectomy were studied. MAIN OUTCOME MEASURES: Vocal cord paralysis after thyroidectomy. RESULTS: There were 213 unilateral and 287 bilateral procedures, with 787 nerves at risk of injury. Thirty-three patients (6.6%) developed postoperative unilateral cord paralysis, and 5 (1.0%) had recognizable nerve damage during the operations. Complete recovery of vocal cord function was documented in 26 (93%) of 28 patients. The incidence of temporary and permanent cord palsy was 5.2% and 1.4% (3.3% and 0.9% of nerves at risk), respectively. Among factors analyzed, surgery for malignant neoplasm and recurrent substernal goiter was associated with an increased risk of permanent nerve palsy. Primary operations for benign goiter were associated with a 5.3% and 0.3% incidence (3.4% and 0.2% of nerves at risk) of transient and permanent nerve palsy, respectively. CONCLUSIONS: Unrecognized recurrent laryngeal nerve palsy occurred after thyroidectomy. Thyroid surgery for malignant neoplasms and recurrent substernal goiter was associated with an increased risk of permanent recurrent nerve damage. Postoperative vocal cord dysfunction recovered in most patients without documented nerve damage. PMID- 10668883 TI - Prevalence and mechanisms of small intestinal obstruction following laparoscopic abdominal surgery: a retrospective multicenter study. French Association for Surgical Research. AB - HYPOTHESIS: The prevalence and mechanisms of intestinal obstruction following laparoscopic abdominal surgery have not been studied extensively. DESIGN: Retrospective review of cases of intestinal obstruction after laparoscopic surgery. SETTING: Sixteen surgical units performing laparoscopy in France. PATIENTS: Twenty-four patients with intestinal obstruction. MAIN OUTCOME MEASURES: Prevalence values and descriptive data. RESULTS: The 3 most frequent primary procedures responsible for intestinal obstruction were cholecystectomy (10 cases), transperitoneal hernia repair (5 cases), and appendectomy (4 cases). Prevalences of early postoperative intestinal obstruction after these procedures were 0.11%, 2.5%, and 0.16%, respectively. Intestinal obstruction was due to adhesions or fibrotic bands in 12 cases and to intestinal incarceration in 11 cases. Obstruction was located at the trocar site in 13 cases (9 incarcerations and 4 adhesions), mainly at the umbilicus, and in the operative field in 10 cases (2 incarcerations in a wall defect after transperitoneal inguinal hernia repair, 4 adhesions, and 4 fibrotic bands). The small intestine was involved in 23 of 24 cases; the other was due to cecal volvulus following unrecognized intestinal malrotation. Intestinal obstruction was treated by laparoscopic adhesiolysis in 6 patients and by laparotomy in 18 patients, 6 of whom required small intestine resection. Three postoperative complications but no deaths occurred. CONCLUSION: Intestinal obstruction following laparoscopic abdominal surgery can occur irrespective of the type of operation; the prevalence is as high as (cholecystectomy and appendectomy) or even higher than (transperitoneal hernia repair) that seen in open procedures. PMID- 10668884 TI - Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma. AB - BACKGROUND: Patients with hepatocellular carcinoma (HCC) often develop recurrences after curative resection or liver transplantation. Therefore, tumor cell dissemination must have occurred preoperatively or intraoperatively. Current staging methods cannot reliably detect micrometastasis. Reverse transcription polymerase chain reaction (RT-PCR) for alpha-fetoprotein (AFP) has been used to detect circulating liver cancer cells, but results in blood samples have been contradictory. HYPOTHESIS: AFP-RT-PCR is a specific and sensitive assay for the detection of disseminated tumor cells in central venous blood and bone marrow samples of patients with HCC and has prognostic relevance. DESIGN: Prospective consecutive series. SETTING: University hospital. PATIENTS AND METHODS: We performed preoperative, intraoperative, and postoperative analyses of central venous blood samples and preoperative analysis of bone marrow samples of patients with HCC and patients without malignant disease, using a modified AFP-RT-PCR method. Preoperative serum AFP levels were measured. Clinical follow-up ranged from 4 to 20 months. MAIN OUTCOME MEASURES: Sensitivity and specificity of AFP-RT PCR, correlation of AFP-RT-PCR results to tumor stage and tumor recurrence. RESULTS: In serial dilution experiments, 50 AFP-expressing HepG2 cells were detected in 10 mL of blood. Peripheral blood samples of 20 healthy volunteers and bone marrow samples of 21 patients with benign diseases consistently tested negative for AFP, whereas 4 of 24 patients with HCC showed AFP expression in bone marrow samples. All these patients had advanced disease; however, correlation of positive RT-PCR results to tumor stage was not significant (P = .07). One of the 4 AFP-positive patients developed an intrahepatic recurrence soon after liver transplantation. Central venous blood of patients with HCC (n = 24) and patients with benign liver diseases (n = 13) equally demonstrated AFP-expressing cells. There was no correlation of RT-PCR results to serum AFP levels. CONCLUSIONS: Perioperative screening for micrometastasis in bone marrow of patients with HCC is sensitive and specific with AFP-RT-PCR and may have prognostic relevance. Alpha-fetoprotein is not a suitable marker for the detection of tumor cells in central venous blood samples. PMID- 10668886 TI - Surgery in Mexico. PMID- 10668885 TI - Maximal human neutrophil priming for superoxide production and elastase release requires p38 mitogen-activated protein kinase activation. AB - HYPOTHESIS: Neutrophil priming has been implicated in the development of multiple organ failure, although the precise intracellular mechanisms that regulate neutrophil priming remain unclear. Our previous work characterized platelet activating factor (PAF) priming of human neutrophils for concordant superoxide anion (O2-) generation and elastase degranulation. The p38 mitogen-activated protein kinase (MAPK) is activated by PAF stimulation. We hypothesized that PAF induced human neutrophil priming for O2- and elastase release is mediated via the p38 MAPK pathway. DESIGN: Isolated neutrophils from 6 human donors were preincubated with the specific p38 MAPK inhibitor SB 203580 (1 micromol/L) or buffer (control) for 30 minutes. Cells were then primed with PAF (200 nmol/L), followed by receptor-dependent (N-formyl-methionyl-leucyl-phenylalanine, 1 micromol/L) or receptor-independent phorbol myristate acetate (PMA, 100 ng/mL) activation. SETTING: Urban trauma research laboratory. PATIENTS: Healthy volunteer donors of neutrophils. MAIN OUTCOME MEASURES: Maximal rate of O2- generation was measured by superoxide dismutase-inhibitable reduction of cytochrome c and elastase release by the cleavage of N-methoxysuccinyl-Ala-Ala Pro-Val-p-nitroanilide. RESULTS: SB 203580 significantly attenuated the generation of O2- and release of elastase from neutrophils activated with N formyl-methionyl-leucyl-phenylalanine but not with PMA. Independent of the activator receptor status, SB 203580 almost completely blocked the exaggerated neutrophil cytotoxic response due to PAF priming. CONCLUSIONS: The p38 MAPK pathway is required for maximal PAF-induced neutrophil priming for O2- production and elastase degranulation. Therefore, the MAPK signaling cascade may offer a potential therapeutic strategy to preempt global neutrophil hyperactivity rather than attempt to nullify the end products independently. PMID- 10668887 TI - Robotics and allied technologies in endoscopic surgery. AB - Endoscopic surgery was developed in the 1970s and 1980s, with initial work conducted by pioneering surgeons. After the development of laparoscopic cholecystectomy, the breakthrough of endoscopic surgery had a great effect on all surgical specialties. Starting with rather simple procedures, such as cholecystectomy, a rapid progression toward more complex procedures, such as reflux or colonic surgery, took place. It was realized at this time that the existing endoscopic instruments allowed only a limited preciseness when performing the procedures, and part of the information from inside the abdominal cavity was not available to the surgeon. This prompted a discussion with engineers concerning the development of more advanced technologies to give those performing endoscopic surgery the same quality of information and manipulation that surgeons have when performing open surgery. These qualities include (1) instruments and manipulators that allow surgical action under endoscopic control with all degrees of freedom; (2) devices that provide surgeons with tactile feedback; and (3) vision systems that provide surgeons with the same quality of visual information as with open surgery, namely, high resolution, excellent color quality, precise spatial information, and a constant clear view for optimal surgical action. At the end of 1999, some of the aforementioned quality concepts found their way into the surgical routine, but most of the concepts are still being developed. Another decade will pass before endoscopic surgery procedures will be closer to the technological goals. PMID- 10668888 TI - Surgical treatment of postoperative incisional hernias by intraperitoneal insertion of a Dacron mesh and an aponeurotic graft. PMID- 10668889 TI - A miracle and a privilege. PMID- 10668890 TI - Internationalization of American surgery. PMID- 10668891 TI - CD40L is critical for protection from demyelinating disease and development of spontaneous remyelination in a mouse model of multiple sclerosis. AB - Theiler's murine encephalomyelitis virus (TMEV) induces acute neuronal disease followed by chronic demyelination in susceptible strains of mice. In this study we examined the role of a limited immune defect (deletion or blocking of CD40 ligand [CD40L]) on the extent of brain disease, susceptibility to demyelination, and the ability of demyelinated mice to spontaneously remyelinate following TMEV infection. We demonstrated that CD40L-dependent immune responses participate in pathogenesis in the cerebellum and the spinal cord white matter but protect the striatum of susceptible SJL/J mice. In mice on a background resistant to TMEV induced demyelination (C57BL/6), the lack of CD40L resulted in increased striatal disease and meningeal inflammation. In addition, CD40L was required to maintain resistance to demyelination and clinical deficits in H-2b mice. CD40L-mediated interactions were also necessary for development of protective H-2b-restricted cytotoxic T cell responses directed against the VP2 region of TMEV as well as for spontaneous remyelination of the spinal cord white matter. The data presented here demonstrated the critical role of this molecule in both antibody- and cell mediated protective immune responses in distinct phases of TMEV-mediated pathology. PMID- 10668892 TI - Fas/CD95/APO-1 can function as a death receptor for neuronal cells in vitro and in vivo and is upregulated following cerebral hypoxic-ischemic injury to the developing rat brain. AB - Fas/CD95/Apo-1 is a cell surface receptor that transduces apoptotic death signals following activation and has been implicated in triggering apoptosis in infected or damaged cells in disease states. Apoptosis is a major mechanism of neuronal loss following hypoxic-ischemic injury to the developing brain, although the role of Fas in this process has not been studied in detail. In the present study, we have investigated the expression and function of Fas in neuronal cells in vitro and in vivo. Fas was found to be expressed in the 14 day old rat brain, with strongest expression in the cortex, hippocampus and cerebellum. Cross-linking of Fas induced neuronal apoptosis both in neuronal PC12 cells in culture and following intracerebral injection in vivo, indicating that neuronal Fas was functional as a death receptor. This death was shown to be caspase dependent in primary neuronal cultures and was blocked by the selective caspase 8 inhibitor IETD. Finally, cerebral hypoxia-ischemia resulted in a strong lateralised upregulation of Fas in the hippocampus, that peaked six to twelve hours after the insult and was greater on the side of injury. These results suggest that Fas may be involved in neuronal apoptosis following hypoxic-ischemic injury to the developing brain. PMID- 10668893 TI - Creutzfeldt-Jakob disease: Carnoy's fixative improves the immunohistochemistry of the proteinase K-resistant prion protein. AB - The neuropathological diagnosis of Creutzfeldt-Jakob disease relies on the immunohistochemical demonstration of the proteinase-K resistant form of the prion protein (PrPres) in the brain tissue. The antigenicity of PrPres is strongly reduced by the formalin solution widely used to fix the tissue, thus the PrPres immunoreactivity is inconsistently detectable in formalin-fixed tissue. A better PrPres immunostaining can be obtained by using Carnoy's fixing solution, which is composed of ethanol, chloroform and acetic acid (6:3:1). PrPres can easily be extracted from Carnoy's-fixed, paraplast-embedded tissue. Accordingly, Carnoy's fixed tissue can prior to immunolabeling be subjected to proteinase K and guanidine thiocyanate, which respectively eliminate the normal cellular form of prion protein and promote protein denaturation. In comparison with the best protocols for formalin-fixed tissue (i.e.--hydrolytic autoclaving or autoclaving in distilled water followed by formic acid and guanidine thiocyanate), PrPres immunostaining carried out on sections cut from Carnoy's-fixed, paraplast embedded tissue blocks and subjected to proteinase K and guanidine thiocyanate, proved more successful to detect and map both diffuse and focal PrPres immunoreactivity, and to correlate the immunoreactivity pattern with MV polymorphism at PRNP codon 129 and PrPres banding and glycosylation pattern revealed by Western blot. PMID- 10668894 TI - Distribution of Borna disease virus in the brain of rats infected with an obesity inducing virus strain. AB - Experimental infection of Lewis rats with Borna disease virus (BDV), a nonsegmented, single-stranded RNA virus, usually causes an immune-mediated biphasic neurobehavioral disorder. Such animals develop a persistent infection of the CNS with viral antigen expression in all brain regions and a disseminated nonpurulent meningoencephalitis. Interestingly, intracerebral infection of Lewis rats with a BDV-variant (BDV-ob) causes a rapid increase of body weight with the development of an obesity syndrome without obvious neurological signs. The obese phenotype is correlated with a characteristic distribution of inflammatory lesions and BDV-antigen in the rat brain. Infiltration with mononuclear immune cells and viral antigen expression are restricted to the septum, hippocampus, amygdala and ventromedian tuberal hypothalamus. Therefore, infection with the obesity-inducing BDV-ob results most likely in neuroendocrine dysregulations leading to the development of an obesity syndrome. This might be due to the restriction of viral antigen expression and inflammatory lesions to brain areas which are involved in the regulation of body weight and food intake. The BDV induced obesity syndrome represents a model for the study of immune-mediated neuroendocrine disorders caused by viral infections of the CNS. PMID- 10668895 TI - Pineal parenchymal tumors: a correlation of histological features with prognosis in 66 cases. AB - The WHO classification of CNS tumors divides pineal parenchymal tumors (PPT) into pineocytoma (PC), pineoblastoma (PB) and mixed pineocytoma-pineoblastoma or PPT with intermediate differentiation. The reported incidence of mixed/intermediate PPT varies and this may reflect the difficulty in classifying tumors of this type. In an attempt to overcome the problem of the classification of PPT with intermediate differentiation, we describe the relationship between histological features and patient survival in a large cooperative series of 66 PPT from 12 neurosurgical centres. All tumors were studied with both light microscopy and immunohistochemically using antibodies against glial markers or neural/neuroendocrine markers. Our series included 11 PC, 39 mixed/intermediate PPT and 16 PB. A number of mitoses greater than 6 and the presence of necrosis were associated with a poorer outcome, while positive immunostaining for neurofilaments was associated with a better survival. We propose a new prognostic grading of 4 grades, grade I for PC, grade II for PPT with fewer than 6 mitoses and positive immunolabelling for neurofilaments, grade III for PPT with either 6 or more than 6 mitoses or fewer than 6 mitoses but without immunostaining for neurofilaments and grade IV for PB. PMID- 10668896 TI - Resuscitative hypothermia protects the neonatal rat brain from hypoxic-ischemic injury. AB - The effect of 24 h of hypothermic recovery on moderate hypoxic-ischemic brain damage in P7-rats was investigated for 42 d after the insult, using magnetic resonance and histopathology. Occlusion of right common carotid artery and 90 min exposure to 8% O2 at 37 degrees C body temperature produced cytotoxic edema of 51(+/-11)% brain volume (BV) and depression of brain energy metabolism (PCr/Pi) from 1.43(+/-0.21) to 0.14(+/-0.11). During recovery, the body temperature was reduced to 30 degrees C for 24 h in 36 animals, but was kept at 37 degrees C in 34 animals. The edema waned upon reoxygenation leaving only the core lesion at 2 h, but reappeared reaching a maximal extent of 11+/-8% BV under hypothermia compared to 45(+/-10)% under normothermia at around 24 h. PCr/Pi recovered transiently within 13 h and declined again to 1.07(+/-0.19) under hypothermia and to 0.48(+/-0.22) under normothermia at around 24 h. Hypothermia led to significant long term brain protection, leaving permanent tissue damage of 12(+/ 6)% BV compared to 35(+/-12)% BV under normothermia. However, animals with severe initial injury developed large infarctions, despite hypothermic treatment. Even then, the time to develop infarction was significantly prolonged, leaving the opportunity for additional therapeutic intervention. PMID- 10668897 TI - Characteristic chromosomal imbalances in primary central nervous system lymphomas of the diffuse large B-cell type. AB - We performed a genome wide screening for genomic alterations on a series of 19 sporadic primary central nervous system lymphomas (PCNSL) of the diffuse large B cell type by comparative genomic hybridization (CGH). The tumors were additionally analyzed for amplification and rearrangement of the BCL2 gene at 18q21 as well as for mutation of the recently cloned BCL10 gene at 1p22. Eighteen tumors showed genomic imbalances on CGH analysis. On average, 2.1 losses and 4.7 gains were detected per tumor. The chromosome arm most frequently affected by losses of genomic material was 6q (47%) with a commonly deleted region mapping to 6q21-q22. The most frequent gains involved chromosome arms 12q (63%), 18q and 22q (37% each), as well as 1q, 9q, 11q, 12p, 16p and 17q (26% each). High-level amplifications were mapped to 9p23-p24 (1 tumor) and to 18q21-q23 (2 tumors). However, PCR-based analysis, Southern blot analysis and high-resolution matrix CGH of the BCL2 gene revealed neither evidence for amplification nor for genetic rearrangement. Mutational analysis of BCL10 in 16 PCNSL identified four distinct sequence polymorphisms but no mutation. Taken together, our data do not support a role of BCL2 rearrangement/amplification and BCL10 mutation in PCNSL but indicate a number of novel chromosomal regions that likely carry yet unknown tumor suppressor genes or proto-oncogenes involved in the pathogenesis of these tumors. PMID- 10668898 TI - Low frequency of SV40, JC and BK polyomavirus sequences in human medulloblastomas, meningiomas and ependymomas. AB - Several reports have suggested a role for polyomaviruses in the pathogenesis of human brain tumors. This potential involvement is not conclusively resolved. For the present study, a highly sensitive PCR-assay with fluorescence-labelled primers was developed to search for polyomavirus sequences in human brain tumor and control DNA samples. The assay was shown to detect approximately one viral large T-antigen (TAg) gene per 250 cells. We identified simian virus 40 (SV40) like sequences in 2/116 medulloblastomas, in 1/131 meningiomas, in 1/25 ependymomas and in 1/2 subependymomas. A single case of ependymoma contained SV40 VP-1 late gene sequences. Moreover, one of the meningioma samples showed JC virus sequences. In contrast, 60 hepatoblastoma samples and 31 brain samples from schizophrenic patients were consistently negative. BK virus sequences were not detectable in any of our samples. Immunohistochemical analysis of two SV40 positive tumor biopsies failed to detect large TAg in the tumor cells. In the JC positive meningioma, immunoreactivity for the viral late gene product (VP-1) was not observed. Our data do not entirely rule out SV40 and JC virus as an initiative agent with a hit-and-run mechanism. However the low frequency of virus sequences and the absence of TAg protein expression argue against a major role of these viruses in the pathogenesis of human medulloblastomas, meningiomas and ependymomas. PMID- 10668899 TI - Introduction: the role of inflammation in mediating damage following stroke and neurotrauma. PMID- 10668900 TI - Inflammation and stroke: putative role for cytokines, adhesion molecules and iNOS in brain response to ischemia. AB - Ischemic stroke is a leading cause of death and disability in developed countries. Yet, in spite of substantial research and development efforts, no specific therapy for stroke is available. Several mechanism for neuroprotection have been explored including ion channels, excitatory amino acids and oxygen radicals yet none has culminated in an effective therapeutic effect. The review article on "inflammation and stroke" summarizes key data in support for the possibility that inflammatory cells and mediators are important contributing and confounding factors in ischemic brain injury. In particular, the role of cytokines, endothelial cells and leukocyte adhesion molecules, nitric oxide and cyclooxygenase (COX-2) products are discussed. Furthermore, the potential role for certain cytokines in modulation of brain vulnerability to ischemia is also reviewed. The data suggest that novel therapeutic strategies may evolve from detailed research on some specific inflammatory factors that act in spatial and temporal relationships with traditionally recognized neurotoxic factors. The dual nature of some mediators in reformatting of brain cells for resistance or sensitivity to injury demonstrate the delicate balance needed in interventions based on anti-inflammatory strategies. PMID- 10668901 TI - Inflammatory mediators of cerebral endothelium: a role in ischemic brain inflammation. AB - Brain inflammation has been implicated in the development of brain edema and secondary brain damage in ischemia and trauma. Adhesion molecules, cytokines and leukocyte chemoattractants released/presented at the site of blood-brain barrier (BBB) play an important role in mobilizing peripheral inflammatory cells into the brain. Cerebral endothelial cells (CEC) are actively engaged in processes of microvascular stasis and leukocyte infiltration by producing a plethora of pro inflammatory mediators. When challenged by external stimuli including cytokines and hypoxia, CEC have been shown to release/express various products of arachidonic acid cascade with both vasoactive and pro-inflammatory properties, including prostaglandins, leukotrienes, and platelet-activating factor (PAF). These metabolites induce platelet and neutrophil activation and adhesion, changes in local cerebral blood flow and blood rheology, and increases in BBB permeability. Ischemic CEC have also been shown to express and release bioactive inflammatory cytokines and chemokines, including IL-1beta, IL-8 and MCP-1. Many of these mediators and ischemia in vitro and in vivo have been shown to up regulate the expression of both selectin and Ig-families of adhesion molecules in CEC and to facilitate leukocyte adhesion and transmigration into the brain. Collectively, these studies demonstrate a pivotal role of CEC in initiating and regulating inflammatory responses in cerebral ischemia. PMID- 10668902 TI - Leukocyte recruitment and the acute inflammatory response. AB - Leukocyte recruitment is a hallmark feature of the inflammatory response. This review summarizes the generally accepted paradigm of leukocyte recruitment based on studies using intravital microscopy to visualize the microcirculation. The role of selectins and alpha4-integrin in rolling as well as integrin-mediated adhesion is discussed. However, it is becoming increasingly clear that the recruitment cascade within organs differs and therefore the review also attempts to highlight what is and is not known regarding leukocyte recruitment into the brain microvasculature. In the second part of this review, we provide some discussion of mechanisms by which the inflammatory response may be terminated. Particular emphasis on nuclear factor Nf kappaB and how IL10, IL13 and secreted leukocyte protease inhibitor (SLPI) may impact upon the Nf kappaB-dependent inflammatory response is presented. PMID- 10668903 TI - The initiation of the microglial response. AB - The initial response of microglia to ischemia and ischemia-like conditions was analyzed in situ and in vitro. After sublethal ischemia in situ, microglia appear activated morphologically, but do not express macrophage-like antigens. In contrast, neuronal damage induces full expression of immunomolecules in microglia. Additionally, blood-borne cells readily infiltrate the region of the ischemic core and constitute another source of cells with macrophage-like expression. Thus, it appears that the microglia are the earliest cells to respond to injury, but their response is graded and complicated by the presence of blood borne immune cells. In vitro ischemia-like conditions caused an irreversible depolarization, ion channel shutdown, and blebbing, indicating that microglia are not equipped to withstand an ischemic insult. Application of ATP alone to microglia produced outward currents and calcium transients and these calcium transients increased when ATP was applied in combination with high potassium. It is known that both outward currents and calcium transients are induced during spreading depression, a feature of focal injury, and this suggests that spreading depression might be one of the initial activators of microglia. PMID- 10668904 TI - Temperature modulation (hypothermic and hyperthermic conditions) and its influence on histological and behavioral outcomes following cerebral ischemia. AB - Core temperature (T(C)) is a critical determinant of the severity of neural damage that results from focal or global ischemia. Former studies indicated that especially intra-ischemic but also post ischemic mild hypothermia significantly decreased necrotic neural damage of a focal or global insult, as assessed between 3-7 days post-insult. More recent work shows that prolonged post-ischemic hypothermia reduces neural damage and inhibits associated behavioral deficits for up to one year after the insult (i.e. true neuroprotection with behavioral preservation). Alternatively, increases in core temperature via external heating or with pyrogens resulting from bacterial infections, at the time of the global ischemia insult worsen the neural damage of ischemic animals from those of respective normothermic controls given the same insult. This is paralleled in the clinical setting whereby approximately 50% of ischemic patients develop fevers within 2 days of the insult and have worsened neurological outcomes than non febrile patients. The review discusses the possible mechanisms of neuroprotection of hypothermic therapy from cerebral ischemia as well as mechanisms involved in the exacerbation of neural damage of hypoxic ischemia under hyperthermic conditions. Questions are raised as to whether the medical community has sufficient evidence to begin appropriate hypothermic therapy of acute stroke patients. The importance of accurate monitoring core temperatures of all suspected stroke patients is emphasized, noting the differences in temperature that can occur with age, sex, medication or lifestyle so that appropriate temperature treatment could be implemented, if required. PMID- 10668905 TI - Redox regulation of nuclear factor kappa B: therapeutic potential for attenuating inflammatory responses. AB - Nuclear factor kappa B (NF-kappaB) is a protein transcription factor that is required for maximal transcription of a wide array of pro-inflammatory mediators that are involved in the pathogenesis of stroke. The purpose of this review article is to describe what is known about the molecular biology of NF NF-kappaB and to review current understanding of the interaction between reactive oxygen species (ROS) in NF-kappaB. ROS seem to play a duel role by participating in the NF-kappaB activation cascade and by directly modulating DNA binding affinity. Exogenous and endogenous antioxidants are effective in blocking activation of NF kappaB and preventing the consequences of pro-inflammatory gene expression. Phase II enzymes either directly or indirectly play a major in vivo role in minimizing oxidative stress by scavenging peroxides, peroxide breakdown products and dicarbonyls and in regeneration of lipid peroxidation chain-breaker, vitamin E. Dietary phase II enzyme inducers have been demonstrated to increase phase II enzyme activities in a variety of tissues. These data, together, suggest that phase II enzyme inducers could have therapeutic value for ameliorating inflammatory conditions. PMID- 10668906 TI - July 1999: 66 year old man with a subacute multi-system disorder and polyneuropathy. PMID- 10668907 TI - August 1999: 49 year old woman with visual disturbances. PMID- 10668908 TI - September 1999: 24 year old female with progressive lower extremity dystonia, dysarthia, dysphagia and mental impairment. PMID- 10668909 TI - October 1999: 70 year old female with seizures and progressive dementia. PMID- 10668910 TI - November 1999: 56 year old woman with left facial numbness. PMID- 10668911 TI - December 1999: 71 year old woman with progressive sensorimotor polyneuropathy. PMID- 10668912 TI - The role of TNF-alpha in human adipose tissue: prevention of weight gain at the expense of insulin resistance? AB - Since evidence has appeared that tumor necrosis factor-alpha (TNF) is involved in the loss of body fat in the course of wasting diseases, a large number of studies have investigated the physiological role of this cytokine in adipose tissue. TNF treatment of several in vitro models of adipogenesis clearly showed that TNF is a potent inhibitor of adipose differentiation. This antiadipogenic property is accompanied by suppression of developmental and metabolic markers of fat cell differentiation, such as peroxisome proliferator-activated receptor (PPAR) gamma2, lipoprotein lipase (LPL), glycerol-3-phosphate dehydrogenase (GPDH) and GLUT4. Moreover, TNF promotes lipolysis in mature adipocytes and, subsequently, a reversion of the adipocyte phenotype. Recent studies demonstrated that TNF directly interferes with the insulin signaling cascade at early steps and, thus, impairs insulin-stimulated glucose transport. Further progress in understanding the role of TNF in adipose tissue was made when endogenous TNF mRNA expression was demonstrated in adipose tissue. Obesity was found to represent a state of overexpression of the TNF system. Such findings support the hypothesis that TNF is a mediator of obesity-linked insulin resistance. However, this concept is mainly based on animal data and is so far only partially supported by studies in humans. Taken together, the results of a variety of experimental and clinical studies suggest that TNF may act as an important auto/paracrine regulator of fat cell function which serves to limit adipose tissue expansion, probably by inducing insulin resistance which may in turn cause metabolic disturbances. Elucidation of the molecular mechanisms of TNF production and action in adipose tissue may help to find new approaches for the treatment of insulin resistance in humans. PMID- 10668913 TI - Stimulation of cardiac glucose transport by thioctic acid and insulin. AB - Thioctic acid (alpha-lipoic acid) has been shown to improve insulin-regulated glucose disposal in animal models of insulin resistance and type 2 diabetic patients. In the present study, we have used isolated adult ventricular cardiomyocytes in order to analyze 1) direct effects of this compound on glucose uptake in a primary muscle cell, and 2) the interaction with the insulin signalling cascade. Both insulin and thioctic acid (2.5 mM) induced a rapid increase in 3-O-methylglucose transport to 322+/-43 and 385+/-58 (n = 5) percent of basal control, respectively. Combined stimulation did not result in an additional significant increase in the transport rate. Preincubation of cardiomyocytes with the phosphatidylinositol 3-kinase inhibitor wortmannin completely abolished the effects of insulin and thioctic acid, whereas gamma linolenic acid selectively blocked the effect of this compound. These data show that thioctic acid mimics insulin action by activating the signalling cascade at or before the level of phosphatidylinositol 3-kinase. PMID- 10668914 TI - Development of low-dose streptozotocin-induced diabetes in ICAM-1-deficient mice. AB - Multiple injections of low-dose streptozotocin (LDSZ) induce immune-mediated insulitis and diabetes in C57BL/6 (H-2b) mice. To evaluate the role of the intercellular adhesion molecule-1 (ICAM-1) for LDSZ induced immune-mediated diabetes, we have investigated mice genetically deficient in the ICAM-1 gene (ICAM-1-/-) in comparison to wild-type (ICAM-1+/+) mice. ICAM-1-/- mice, which had a mixed genetic background of C57BL/6 and DBA/2 mice, were backcrossed to C57BL/6 mice and screened for H2b homogenicity. Mice received five daily injections of 40 mg/kg streptozotocin. On day 21 after the first LDSZ injection 55% of the ICAM-1+/+ (female 33%, male 80%) and 50% of the ICAM-1-/- (female 20%, male 100%), mice had blood glucose levels over 200 mg/dl. Mean blood glucose levels increased in response to LDSZ treatment, however, no differences between ICAM-1+/+ and ICAM-1-/- mice were noted. Histological examinations of pancreatic islets revealed mononuclear infiltration of pancreatic islets without significant differences between both groups of mice. In summary, LDSZ-induced immune-mediated insulitis and diabetes development occurs in ICAM-1-/- mice similarly than in ICAM-1+/+ mice. These results do not support the hypothesis that ICAM-1 plays a key role during immune-mediated infiltration and destruction of pancreatic islets in LDSZ induced diabetes. PMID- 10668915 TI - Potent beta-cell protection in vitro by an isoquinolinone-derived PARP inhibitor. AB - Activation of the nuclear enzyme poly(ADP-ribose)polymerase (PARP) is a critical step in beta-cell death in response to exposure with free radicals or other DNA damaging agents. Nicotinamide, a B vitamin, exerts its beta-cell protective action primarily via its ability to block excessive PARP activity. We show here that the isoquinolinone derivative PD128763, a specific PARP inhibitor, provides protection from cell death in islet cells exposed in vitro to nitric oxide or oxygen radical generating compounds or to the beta-cell toxin streptozotocin, at concentrations 100 times less than required for nicotinamide. Furthermore, while the protective action of nicotinamide is rapidly lost after washing of islet cells, the effects of PD128763 are more long lasting. Both compounds had little capacity to rescue damaged islet cells from subsequent lysis. We conclude that the isoquinolinone derivative PD128763 is superior to nicotinamide in enhancing the resistance of beta-cells towards inflammatory attacks. PMID- 10668916 TI - Diabetes mellitus induces long lasting changes in the glucose transporter of rat heart endothelial cells. AB - The accumulation of glucose exerts various cytotoxic effects on endothelial and other vascular cells, and thereby contributes to the development of microvascular complications in diabetes. Since tissues, in which vascular complications typically occur, do not take up glucose in an insulin regulated manner, it is an important question to know whether other mechanisms exist in these cells to restrict the uptake and the accumulation of glucose. To study this question, we used microvascular endothelial cells isolated from rat heart endothelial cells (RHEC). In RHEC, the non-insulin regulated glucose transporter (Glut-1) was detected as a broad protein band of 50-65 kD. In contrast, the Glut-1 from rat brain, which was taken as reference, had a molecular weight of 45 kD. After treatment with endoglycosidase F, both proteins formed a band of approximately 40 kD on SDS-PAGE, demonstrating a more extensive glycosylation of Glut-1 in RHEC as compared to brain. Incubation of the cells in high glucose (22 mM, up to 10 days) did not down-regulate either Glut-1 protein or mRNA. In contrast to high glucose, deprivation of the cells from glucose led to an increase in Glut-1 mRNA and protein which is partly non-glycosylated. In cells from hearts of streptozotocin diabetic rats (DRHEC), Glut-1 protein, but not Glut-1 mRNA, was reduced by about 40%. Additionally, a significant amount of glycosyl residues was resistant to the enzymatic treatment with N-endoglycosidase F. Both changes in Glut-1 were also observed when the cells were cultivated in low glucose (5.5 mM) for several passages indicating a long lasting, hardly reversible modification of Glut-1 by diabetes. These data indicate that Glut-1 is not down-regulated in RHEC by high glucose, and that this important mechanism to protect the endothelium against an intracellular accumulation of glucose is missing in RHEC. As a consequence, increases in blood glucose may lead to a glucose overload with the described deleterious effects on the structure and function of endothelium. PMID- 10668917 TI - Effect of 1alpha,25(OH)2-vitamin D3 on TNF alpha-mediated apoptosis of human primary osteoblast-like cells in vitro. AB - 1alpha,25(OH)2-vitamin D3 is a hormone which potentially stimulates bone cell growth and differentiation. TNFalpha is one possible inductor for apoptosis; apoptosis being an important regulatoring factor for bone modelling and remodelling. We examined the influence of physiological levels (0.1 nM) 1alpha,25(OH)2-vitamin D3 on TNFalpha-mediated apoptosis in human osteoblast-like cells. These human cells were obtained from bone fragments obtained during orthopedic operations on patients without systemic bone disease. Treatment with 1alpha,25(OH)2-vitamin D3 for 8 weeks resulted in a significant reduction (30%) of viable cell number compared to untreated cells. Incubation with TNFalpha (100 ng/ml for 4 hours) only had limited effects on the rate of apoptosis in control cells. After pretreatment with 1alpha,25(OH)2-vitamin D3, induction of apoptosis increased up to 10% in human osteoblast-like cells. In parallel to the induction of apoptosis, 1alpha,25(OH)2-vitamin D3 stimulated osteocalcin and alkaline phosphatase as markers of mature osteoblasts. Our data suggest that 1alpha,25(OH)2-vitamin D3 has a stimulatory effect on TNFalpha-induced apoptosis in human osteoblast-like cells as a result of 1alpha,25(OH)2-vitamin D3-induced cell differentiation. PMID- 10668918 TI - High frequency of diabetes-specific autoantibodies in parents of children with type 1 diabetes. DENIS study group. AB - Strategies to identify subjects at risk for type 1 diabetes are largely based on the detection of autoantibodies directed to various beta cell autoantigens. Most previous studies only comprise siblings and children of patients with type 1 diabetes; only scare data are available on the antibody profile in older relatives. In this study, we examined the prevalence of cytoplasmic islet cell antibodies (ICA), antibodies to glutamic acid decarboxylase (GADA), antibodies to the protein tyrosine phosphatase IA-2 (IA-2A) and IA-2beta (IA-2betaA) in 531 unaffected parents of patients with type 1 diabetes, and compared the results with antibody frequencies in 2425 siblings. The frequency of ICA, GADA and IA-2A was substantially higher among siblings as compared to parents of patients with type 1 diabetes (8.0% vs. 4.5%, 8.0% vs. 4.3%, and 4.5% vs. 1.9%, respectively; p<0.01). However, subdividing the probands according to age revealed a high prevalence of ICA (5.5 %), GADA (5.9 %), and IA-2A (3.1%) among parents aged 31 40 years which was similar to that observed in siblings above 20 years of age (6.4%, 6.4%, and 3.1%). In both cohorts, GADA and IA-2A were significantly associated with the presence of ICA. The combined screening for GADA and IA-2A identified 100% of parents and 91.9% of siblings at high risk for type 1 diabetes (>10 JDF-U). Furthermore, the analysis of antibody combinations revealed that among antibody positive individuals the percentage of subjects with two or three antibodies was even higher in parents (69.0%) than in siblings (58.2%). The present study shows a high frequency of single and multiple autoantibodies in unaffected parents of patients with type 1 diabetes. Our data indicate that GAD and IA-2 not only represent the major target of autoantibodies in young siblings but also in adult relatives. These findings may be important for the design of future intervention studies. PMID- 10668919 TI - Dendritic cell immunotherapy induces antitumour response in parathyroid carcinoma and neuroendocrine pancreas carcinoma. AB - Parathyroid carcinomas and neuroendocrine carcinomas of the pancreas are rare malignancies in humans. Because of their low radio- and chemosensibility, they fail to respond to conventional therapy. We therefore tested a dendritic cell immunotherapy in an attempt to control the tumour growth in two patients. Studies on mice and humans have demonstrated the potent capacity of dendritic cells to induce specific antitumour immunity. Mature dendritic cells were generated from peripheral blood monocytes in the presence of granulocyte/macrophage colony stimulating factor, interleukin 4 and tumour necrosis factor alpha. Dendritic cells were either loaded with parathyroid hormone (PTH) or with (pancreas) tumour derived lysate (TL), respectively, and were delivered by subcutaneous injections. All immunizations were well tolerated with no side effects, and were administered on an outpatient basis. After repeated vaccinations, specific in vivo immune response was demonstrated by positive delayed-type hypersensitivity (DTH) toward PTH or TL, demonstrating the efficient generation of antigen-specific memory T cells. DTH reactivity was accompanied by a significant decrease of tumour markers in both patients. This approach might be generally applicable to other advanced, radio- and chemotherapy-resistant endocrine malignancies. PMID- 10668920 TI - High dose supplementation of RRR-alpha-tocopherol decreases cellular hemostasis but accelerates plasmatic coagulation in type 2 diabetes mellitus. AB - BACKGROUND: Diabetes mellitus is associated with increased generation of free oxygen radicals and depleted scavenging potential (oxidative stress), leading to increased LDL oxidation and platelet hyperreactivity, the major components of atherothrombotic vascular lesions. A main goal of antioxidant therapy is to protect the LDL particle from atherogenic oxidation and to reduce the activated cellular hemostasis. METHODS: We evaluated the influence of a high dose supplementation with 800 IU of the natural antioxidant RRR-alpha-tocopherol (vitamin E) per day for six months on serum levels, vitamin E load of LDL particles (HPLC), lag phase of LDL oxidation (Esterbauer's assay), platelet adhesion molecules, leukocyte-platelet coaggregation (flow cytometry, D-III protocol) and coagulation (INR/PTT) in a group of 36 patients with type 2 diabetes (f/m 22/14; age 58+/-8.0; HbA1 at baseline 10.25+/-1.7). RESULTS: Average vitamin E levels increased 2.65-fold accompanied by a 1.83-fold increase of LDL-associated vitamin E and a 12.3 min prolongation of the lag-phase of LDL oxidation (p<0.001 for all parameters at six months). Platelet expression of PECAM-1 (CD31) (-30.2% positive cells, p<0.001; antigen density -25%, p<0.001), ICAM-2 (CD102) (-2.9% positive cells, p<0.01; antigen density -10.6%, p<0.001) and fibrinogen (-1.6% positive cells, p<0.001; antigen density - 16.1 %, p<0.001) decreased. Concomitantly, platelet-leukocyte-coaggregation increased by 44% (p<0.001), correlating to an INR reduction of 10.4% (1.06+/-0.09 to 0.95+/-0.09, p<0.001, r = - 0.34). The PTT remained constant. CONCLUSION: The antioxidant protection from the increased vitamin E was accompanied by a decreased expression of constitutive and function-dependent platelet adhesion molecules. However, increases in platelet-leukocyte coaggregates and a shortened INR time suggest extrinsic coagulation activation, possibly by induction of a leukocyte tissue factor dependent mechanism. High dose supplements of alpha-tocopherol may override the available redox balance in well controlled type 2 diabetes. However, intrinsic effects of alpha-tocopherol must be discussed. PMID- 10668921 TI - Normal ranges and reproducibility of statistical, geometric, frequency domain, and non-linear measures of 24-hour heart rate variability. AB - Diabetic cardiovascular autonomic neuropathy (CAN) carries an increased risk of mortality. The early detection and characterization of CAN has traditionally been based on the results of autonomic reflex tests (AFTs). A variety of different measures to quantify 24-hour heart rate variability (HRV) have recently been introduced, but their normal ranges, reliability, and validity in patients with CAN have not been adequately studied. We established the normal ranges of statistical (SDNN index, CV, SNN50, RMSSD), geometric (triangular index (TI), triangular interpolation (TINN), top angle index [TAI]), frequency domain (spectral power in the VLF, LF, and HF bands, LF/HF ratio, LF in normalized units [NU]), and non-linear measures (CV1 and CV2 of the Poincare plot) of 24-hour HRV in 94 healthy control subjects. Day-to-day reproducibility was evaluated on two occasions in 17 healthy subjects and 9 diabetic patients. The parameters of HRV were computed over time periods representing the day (6:00-24:00 hours), night (00:00-6:00 hours), and 24 hours in total. The results of all indexes, except for the LF/HF ratio and LF-NU, declined significantly with increasing age (p<0.05), but were independent of sex and BMI. The statistical, geometric, and non-linear measures (p<0.05), but not the frequency-domain parameters decreased significantly with increasing heart rate. Since the HRV data showed log normal distribution, log transformation was used to define the age-related lower limits of normal at the 2.5th centile. Intraindividual reproducibility was highest for the geometric measures. The nonlinear and statistical parameters also showed high reliability, except for the SNN50. The repeatability of the frequency domain measures was somewhat lower but still satisfactory. Reproducibility was lower in the diabetic than in the control group, higher during the day than during the night, and better than that reported previously for the AFTs. In conclusion, in healthy subjects the measures of 24-h HRV are not related to sex or BMI, but strongly dependent on age and heart rate, the latter except for the frequency domain measures. The majority of the HRV measures, in particular the geometric parameters, show a relatively high intraindividual reproducibility which underlines their suitability for the use in prospective studies. PMID- 10668922 TI - Prevalence of chronic complications, metabolic control and nutritional intake in type 1 diabetes: comparison between different European regions. EURODIAB Complications Study group. AB - This study compares the prevalence of chronic complications, the quality of metabolic control and the nutritional intake in people with type 1 diabetes in different European regions. The EURODIAB Complications Study included a sample of 3250 European patients with type 1 diabetes stratified for gender, age and diabetes duration. All examinations were performed using standardised, validated methods. HBA1c, LDL-cholesterol and fasting triglycerides were higher in the eastern European centres than in the southern or north-western European centres. Acute (severe ketoacidosis, severe hypoglycaemia) and chronic diabetes complications (retinopathy, nephropathy, neuropathy, cardiovascular disease) were all considerably more frequent in the eastern European centres. HbA1c was lower in the German centres than in the total EURODIAB cohort or in the north-western European centres, but severe hypoglycaemia and proliferative retinopathy were more common. Persons from the eastern European and the German centres consumed undesirably high amounts of cholesterol, total and saturated fat. Overall, improvements in the prevention, detection and management of diabetes complications in persons with type 1 diabetes are essential throughout Europe, particularly in eastern European regions. Since elevated LDL-cholesterol levels and hypertension were strikingly common in this relatively young cohort of European people with type 1 diabetes, generally more attention should be directed towards an adequate management of these cardiovascular risk factors. PMID- 10668923 TI - Detection of autoantibodies to the diabetes-associated antigen IA-2 by a sensitive enzyme-linked immunosorbent assay. AB - The tyrosine phosphatase like protein IA-2 is an important autoantigen in insulin dependent diabetes mellitus (type 1 diabetes). Autoantibodies to IA-2 (IA-2A) are present in the serum of patients with type 1 diabetes even before the onset of the disease. Previously, we reported on a radioimmune assay to detect IA-2A, using E. coli-derived 125I-labelled IA-2 as antigen. Although this assay could be shown to be equivalent to the common reference method for IA-2A detection (radioligand assays using in vitro synthesised 35S-methionine labelled antigen), the disadvantages of both assays with respect to synthesis and handling of the radioactive antigen limit their use in routine laboratories. In this study, we have evaluated a non-radioactive enzyme-linked immunosorbent assay (ELISA) for the simple detection of IA-2A. We report on an ELISA where the biotinylated intracytoplasmic part of IA-2 (IA-2ic) is captured on streptavidin-coated plates. The sensitivity of the ELISA was similar to the validated radioligand assay, as it detected 47 of 69 (69%) patients with type 1 diabetes as compared to 46 of 68 (67 %) with the reference method for IA-2A detection (radioligand assays using in vitro synthesised 35S-methionine labelled antigen). Only 2 of 50 (4%) patients with autoimmune thyroid disease and 1 of 114 (1 %) healthy controls were detected in the ELISA, confirming specificity. There was a significant correlation between the ELISA and the radioligand assay (r = 0.64, p<0.001). We conclude that this ELISA is suitable to detect IA-2A in the serum of patients with type 1 diabetes with a similar sensitivity and specificity to the radioligand assay. This ELISA will allow rapid and simple measurement of IA-2A where the radioligand assay is inconvenient or not available. PMID- 10668924 TI - The malformed kidney: disruption of glomerular and tubular development. AB - Renal malformations are the major cause of renal failure during early childhood. They are found in approximately 100 genetic syndromes. We review the embryologic development of the kidney and its molecular control. Important new information has been derived from mutational analysis in humans and mice. We describe how mutations in nine transcription factors, 12 signaling molecules and nine gene products involved in a variety of other cellular functions disrupt renal morphogenesis. The information presented provides a template for integrating new discoveries on the molecular basis of renal development, for classifying renal malformations observed in the clinical setting, and for identifying defective genes in affected patients. PMID- 10668925 TI - Transcervical cells and the prenatal diagnosis of haemoglobin (Hb) mutations. AB - Prenatal diagnoses of haemoglobin (Hb) mutations were performed using transcervical cells, retrieved by aspiration from the endocervical canal of ten selected pregnant women at about 10 weeks of gestation, prior to chorionic villus sampling (CVS). Both parents were carriers of haemoglobinopathies (thalassaemia or HbS). Clumps of fetal cells were isolated by micromanipulation under an inverted microscope and aliquots of the extracted DNA tested separately for the presence of paternally derived chromosome markers and Hb mutations by quantitative fluorescent polymerase chain reaction (PCR). The correct prenatal diagnosis of Hb diseases, using selected single clumps of trophoblastic cellular elements free of maternal contaminating cells, was achieved in six out of ten cases. PMID- 10668926 TI - Mosaicism for a small marker chromosome resulting from a familial Robertsonian translocation (21;22). AB - A mosaic marker chromosome found in amniotic fluid was shown to have originated from the proximal part of the long arm of chromosome 22. This marker is unusual because it is the result of a deletion of a maternally inherited Robertsonian 21;22 translocation. It is suggested that the deletion and marker formation probably occurred post zygotically in the fetus. This rare case illustrates the difficulty in estimating risk of fetal abnormalities associated with de novo marker chromosomes. In this example, although the 'extra' marker chromosome contains euchromatin, the karyotype may still be 'balanced'. PMID- 10668927 TI - Utility of the predictors of coronary heart disease mortality in a longitudinal study of elderly Finnish men aged 65 to 84 years is dependent on context defined by Apo E genotype and area of residence. AB - A common assumption underlying most genetic studies is that individuals with different genotypes respond similarly to exposure to internal (epigenetic and background genotype effects) and external (ecological) environments. Here we evaluate whether this assumption is true in individuals with different genotypes of the gene coding for the apolipoprotein E (Apo E) molecule, an important determinant of the metabolic fate of plasma lipids and lipoproteins. We addressed whether the utility of known risk factors of coronary heart disease (CHD) in the prediction of CHD death in a 5-year follow-up is the same for the two most common Apo E genotypes, epsilon3/3 and epsilon4/3, in two cohorts of elderly Finnish men (age at baseline: 65-84 years), one in Eastern and the other in Southwestern Finland. The CHD mortality rate was higher in the epsilon4/3 than in the epsilon3/3 genotype in both cohorts (11.1 versus 7.8%, Pr = 0.281 in the Eastern cohort and 19.6 versus 8.2%, Pr = 0.002 in the Southwestern cohort). In the Eastern cohort, serum high density lipoprotein (HDL) cholesterol level was identified as a strong predictor of CHD death in the epsilon3/3 genotype (beta = 2.155, Pr = 0.019). In the Southwestern cohort, age (beta = 0.139, Pr = 0.006), body mass index (BMI) (beta = 0.149, Pr = 0.016), and serum total cholesterol level (beta = 0.453, Pr = 0.051) were identified as strong predictors in the epsilon3/3 genotype, as were smoking (beta = 0.236, Pr = 0.008) and BMI (beta = 0.124, Pr = 0.057) in the epsilon4/3 genotype. The latter observation indicates that in Southwestern Finland the probability of CHD death decreases with increasing BMI in elderly men with the epsilon4/3 genotype, while in their counterparts with the epsilon3/3 genotype the risk increases with increasing BMI. This difference was statistically significant (Pr = 0.002). These observations clearly argue against the assumption that individuals with different genotypes respond similarly to exposures to internal and/or external environments. These observations are consistent with a complex pathobiology of CHD involving biochemical and physiological agents that are under the influence of interactions between genetic and environmental factors. Information about these interactions is necessary for developing a more precise risk assessment and ultimately to improve public health and clinical strategies to prevent this devastating disease both at the individual and population levels. PMID- 10668928 TI - Normolipidemia and hypercholesterolemia in persons heterozygous for the same 1592 + 5G --> A splice site mutation in the low-density lipoprotein receptor gene. AB - In the present study, we have characterized a unique splice donor G to A substitution in the moderately conserved + 5 position in intron 10 of the low density lipoprotein (LDL) receptor gene. In two Danish families, carriers of the 1592 + 5G --> A mutation display a clinical phenotype ranging from healthy normocholesterolemic persons to classical heterozygous familial hypercholesterolemia (FH) patients. Reverse transcription-polymerase chain reaction (RT-PCR) of RNA from Epstein Barr virus (EBV)-transformed lymphoblasts obtained from members of both families demonstrated abnormal splicing generating two aberrant mRNAs due to either alternative splicing and skipping of exon 10 or activation of a cryptic splice site in intron 10 inserting 66 intronic base pairs. These abnormally spliced mRNAs were predicted to encode two abnormal receptor proteins containing an in-frame deletion of 75 amino acids and an insertion of 22 novel amino acids, respectively. Results obtained by immunofluorescence staining, flow cytometry, and confocal microscopy of transfected Chang and COS-7 cells expressing normal and mutant LDL receptors were compatible with nearly complete retention of the mutant proteins in the endoplasmic reticulum. Quantitative measurements of LDL receptor mRNAs from EBV transformed lymphoblasts, however, did not reveal any significant differences in variant mRNA contents between mutation carriers in the families that could be related to degree of hypercholesterolemia. PMID- 10668929 TI - Genetic and segregation analysis of congenital cataract in the Indian population. AB - Congenital cataract is a major cause of blindness in children, and there is wide variation in the few reports available on the frequencies of its different inheritance patterns. Two hundred and fifty-two families with congenital cataract belonging to 13 different states of India, were clinically and genetically investigated to study their inheritance and segregation patterns. Twenty-one percent of the cases were autosomal recessive, 15% autosomal dominant, 63% were simplex cases, and in the remaining cases the inheritance pattern was not clear. A high incidence of consanguinity (50.9%) was observed in autosomal recessive cases. Out of 340 affected individuals, 222 (65.3%) were males and 118 (34.7%) were females. Segregation analysis showed good agreement in autosomal dominant and recessive families and the data are indicative of the prevalence rate for different inheritance patterns of congenital cataract within the Indian population. PMID- 10668930 TI - Association between polymorphism of the cholecystokinin gene and idiopathic Parkinson's disease. AB - Parkinson's disease (PD) is characterized by major alterations of neurotransmitter activity due to damage of the substantia nigra. Changes in neuropeptide concentration within the basal ganglia may play an important role in the putative dopaminergic-peptidergic interactions associated with the disease. Cholecystokinin (CCK) modulates the release of dopamine in the mesolimbic pathway and affects dopamine-related behavior. We analyzed genetic variations in the CCK gene, in both the coding and promoter region, in order to investigate the role of polymorphism in idiopathic PD. Four polymorphic sites of the CCK gene (-196G/A, 45C/T, 1270C/G, 6662C/T) were found in PD patients and controls. Complete linkage disequilibrium was observed between the -45 locus and the 1270 locus, and also a possible linkage disequilibrium was found between the -45 and -196 loci. A significant difference was found in the distributions of three identified genotypes at the -45 locus between 116 PD patients and 95 age-matched control subjects (chi2 = 7.95, p = 0.018, Bonferroni correction; p = 0.054). In addition, a significant difference was obtained amongst the three genotypic groups at the 45 locus when compared between PD patients who experienced hallucinations (n = 23) and those (n = 93) who did not (chi2 = 8.08, p = 0.018, Bonferroni correction, p = 0.126). Our data suggested that mutations at the -45 locus in the promoter region of the CCK gene may influence vulnerability to hallucinations in PD patients treated with L-dopa. PMID- 10668931 TI - Polyvariant mutant CFTR genes in patients with chronic pancreatitis. AB - Several authors have reported an association between mutations of the cystic fibrosis transmembrane conductance regulator gene (CFTR) and chronic pancreatitis. CFTR gene transcription and protein efficiency are influenced by two polymorphic loci, (TG)m and M470V, other than the T5 allele, whose role is already well-established. The TG11/T5 haplotype is commonly found in healthy subjects, while the TG12/T5/V470 and TG13/T5/V470 haplotypes are present in congenital bilateral absence of the vas deferens (CBAVD) patients. While the T5 allele is a mutation that is over-represented in patients with chronic pancreatitis, no data are available concerning the possible allelic preference at the other two polymorphic loci, (TG)m and M470V, in these patients. For this reason, we screened 39 patients with chronic pancreatitis for the most common CFTR mutations found so far in the Italian population; in addition, we examined the length of the polypyrimidine (poly-T) tract in intron 8, the (TG)m length and the M or V codon at position 470. CFTR mutations were found in 3 patients. Poly-T variant typing identified genotype T5/T7 in 5 patients and T5/T9 in 1 patient. Direct sequencing of intron 8 in patients with the T5 variant revealed the TG12/T5/V470//TG11/T7/V470 genotype in 5 patients and TG10/T9//TG11,T5 genotype in 1 patient. In patients with chronic pancreatitis, the T5 allele is frequently associated with TG12 and V470, a haplotype already reported in CBAVD cases and quite uncommon in healthy subjects. PMID- 10668932 TI - Cerebrotendinous xanthomatosis without xanthomas. PMID- 10668933 TI - Role of the Pro23Leu mutation in a family affected by retinitis pigmentosa in the Basque Country. PMID- 10668934 TI - The incidence of tumours in renal transplant recipients with long-term immunosuppressive therapy. AB - INTRODUCTION: The incidence of cancers after renal transplantation is significantly higher than in population that have not undergone transplantation. It is increased by a long-term survival of functional graft requiring long-term immunosuppressive therapy. MATERIAL AND METHODS: Since 1972, 620 renal transplantations have been performed for different causes of end stage renal disease. The authors report a group of 18 renal transplant patients (2.9%) who had cancer. Patients with malignancies are reviewed according to their age, sex, type of immunosuppression, interval between transplantation and the diagnosis of cancer, method of treatment and survival. RESULTS: All patients received cadaver kidneys, and secondary transplantation was performed in two patients. Five patients received conventional immunosuppression--azathioprine with prednisone, another 13 patients received cyclosporine with prednisone and/or azathioprine. In 13 males and 5 females (mean age 46.1 years) the malignant disease developed about 62.4 months after renal transplantation. Six patients had epithelial skin cancers (four of them had squamous cell carcinomas and two basal cell carcinomas). Two patients had breast cancer, colorectal carcinoma, renal cell carcinoma and bladder cancer, respectively, one patient had gastric cancer, thyroid carcinoma, carcinoma of tonsilla, and monocytic leukaemia with blastic transformation, respectively. The average survival of patients with malignancies was 20.3 months. Of 17 patients with cancer, 13 underwent surgical treatment, four patients with advanced disease received radiotherapy, hormonal treatment or only symptomatic therapy. In one patient the malignant disease was only discovered at autopsy. Five patients died of progressive malignant disease, four of intercurrent disease. Nine (50%) patients are alive, with no evidence of disease (NED), 31.9 months in average following the diagnosis of malignancy. Three patients returned to dialysis treatment, other 6 patients live with well functioning graft. CONCLUSIONS: In patients surviving long time after kidney transplantation the possibility of development of malignant disease should be considered. Preventive evaluation should guarantee early detection of cancer. Appropriate treatment, without cessation of immunosuppressive therapy, is indicated with the intention to prolong the patients' life with a functional graft and without dialysis treatment. PMID- 10668935 TI - Idiopathic retroperitoneal fibrosis revisited. AB - We have evaluated 21 patients, ranging in age from 44 to 71 years (mean: 55 years), who presented to our department with the radiologic characteristics of retroperitoneal fibrosis. Ureterolysis was performed in all cases. Intraperitoneal placement of the ureter was performed in 9, and placement of the ureter in a lateral extraperitoneal position in 12 cases. We found no difference in the postoperative course and radiological and clinical improvement to favour the first or the second method, and therefore we consider the intraperitoneal approach as an unnecessary manoeuvre. PMID- 10668936 TI - Glycosaminoglycans excretion in interstitial cystitis. AB - Glycosaminoglycans (GAGs) have been identified histochemically and biochemically in urothelium. They have been suggested to have an anti-adherence effect at the bladder surface important in the urothelial defence against bacterial and carcinogenic insult. The aim of this study was to investigate urinary GAGs excretion in patients with interstitial cystitis. Urinary GAG excretion was determined in patients with interstitial cystitis (n:34; 16 males, 18 females) and healthy subjects (n:34; 16 males, 18 females). Urinary GAG determinations were made by the dimethymethylene blue method. Student's t-test was used for statistical analysis of the results. Urinary GAG excretion was found to be elevated significantly in patients with interstitial cystitis (14.45+/-2.02 mg/g Cr) as compared to healthy subjects (10.11+/-2.3 mg/g Cr) (p<0.01). There was no significant difference in urinary GAG excretion between males and females in either the healthy subjects or in the patients. Determination of urinary GAG excretion may be an important non-invasive test in the investigation of patients with interstitial cystitis. PMID- 10668937 TI - Prognostic significance of p53 protein accumulation in stage pT1 transitional cell carcinoma of the bladder. AB - OBJECTIVE: Mutations in the tumour suppressor gene p53 results in the production of a mutant type, dysfunctional p53 protein which can readily be detected in the cell nucleus by immunohistochemical staining. This study aims to investigate the association of nuclear p53 protein accumulation with the clinical outcome of stage pT1 transitional cell carcinoma of the bladder which is renowned for high rates of recurrence and progression. METHODS: TUR samples of the tumours from fifty-two patients with primary stage T1 bladder cancer were analyzed immunohistochemically using the standard avidin-biotin peroxidase method for nuclear p53 accumulation. Status of p53 immunostaining was correlated with tumour recurrence, disease progression and three-year survival of each patient. RESULTS: The rate of tumour recurrence in pT1 bladder cancer was 36% in patients with tumours stained negatively for p53 protein and 78% in patients with tumours stained positively for p53 protein. Disease progression was seen in 15% of p53 ( ) patients and in 56% of p53 (+) patients. CONCLUSIONS: In stage pT1 bladder tumours p53 nuclear accumulation indicates higher rates of tumour recurrence and disease progression. Accordingly, in patients who have pT1 bladder tumours with nuclear p53 accumulation, institution of more aggressive therapy should be considered and early radical therapeutic modalities should be offered to these patients. PMID- 10668938 TI - Tissue polypeptide antigen and carcinoembryonic antigen lack diagnostic accuracy in urothelial carcinoma. AB - Carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA) levels in serum and urine from 25 patients with bladder cancer and 42 patients with cancer of the renal pelvis/ureter have been evaluated as an aid for clinical diagnosis of urothelial cancer. The tumour CEA content varied markedly, from values obtained in normal urothelium up to 822 and 7306 ng/g wet tissue in cancer of the renal pelvis/ureter and bladder cancer, respectively. Serum and urine CEA levels were found not to correlate with the tumour CEA content. Serum CEA levels were found increased over 5 microg/L in up to 16% of bladder cancer patients, but only in 4.8% in renal pelvis/ureter cancer. Urine of cancer patients contained usually normal CEA levels. Increased serum TPA levels were found in 48% and 35.7% of patients with bladder cancer and cancer of renal pelvis/ureter, respectively. Urine TPA levels were significantly increased in both, patients with bladder cancer (p<0.001) and cancer of renal pelvis/ureter (p<0.01). The median values of urine TPA were 59, 1095 and 1325 U/L, in controls, patients with bladder cancer and cancer of renal pelvis/ureter, respectively. However, considering previously described increase of TPA in inflammatory diseases of urinary tract and in renal failure patients, results of urinary TPA obtained in the diagnostic workup of urothelial cancer should be cautiously interpreted. This study shows that serum and urine levels of CEA and TPA have no diagnostic accuracy required for clinical diagnosis of urothelial cancer. PMID- 10668939 TI - The effect of intravesical mitomycin C on the recurrence of superficial (Ta-T1) bladder cancer. A Hungarian Multicenter Study. AB - We evaluated the prophylactic efficacy of instillations of intravesical mitomycin C in 57 patients with primary superficial bladder cancer in a multicenter clinical trial. After complete transurethral resection of Ta-T1 G1-G2 transitional cell bladder carcinomas, patients were treated with mitomycin 40 mg/50 ml saline of 15 instillations for 12 months. Most of the complications were mild and transient but two patients dropped out of the trial because of moderate side effects. Fifty-one patients were evaluable. We observed tumour recurrences in six patients (11.8%) during a median follow-up of 44.5 months. The recurrences were treated by transurethral resection. There was no muscle invasive progression in the recurrences. Our investigations confirm the effectiveness of mitomycin C in the treatment of patients with superficial bladder cancer. PMID- 10668940 TI - Squamous metaplasia of the bladder: findings in 14 patients and review of the literature. AB - We have documented the data of squamous metaplasia of the bladder in 14 patients who had undergone cystoscopies for different reasons. In two biopsies, there were marked keratinization and cellular atypia. One of these two subjects was diagnosed as squamous carcinoma of the prostate and transitional cell carcinoma of the bladder. The lesions of 8 female patients were on the trigone and evaluated as normal anatomical variants due to hormonal changes. Three of them were remarkable because of recurrent urinary infections. Apart from the two male patients with squamous cell carcinoma of the prostate the two male patients with current squamous metaplasia have been following up. In this study, we have also reviewed the relationship between squamous metaplasia, infection and malignancy. PMID- 10668941 TI - Augmentation ureterocystoplasty with ipsilateral renal preservation in the management of patients with compromised renal function secondary to dysfunctional voiding. AB - We evaluated the role of ureterocystoplasty with ipsilateral renal preservation in the management of patients with neurovesical dysfunction and impaired renal function. The procedure was carried out on 6 patients with a mean age of 8.5 years. All patients had vesicoureteric reflux (VUR) secondary to neuropathic bladders, recurrent urinary tract infections, day time incontinence, impaired and deteriorating renal function. All patients were followed up with a mean of 22.5 months (range 6-30). Renal function stabilized in 4 patients and improved in 2 patients. Adequate urinary bladder capacity was achieved in all patients. Bladder volume increased from a mean of 210+/-71 to 382+/-66, this increase was statistically significant (p<0.001). All patients were dry by day including the children who at presentation were in diapers. We conclude that the results of this operative intervention are satisfactory and promising in the management of this difficult group of patients while avoiding the side effects of enterocystoplasty procedures. PMID- 10668942 TI - Effects of doxazosin in men with benign prostatic hyperplasia: urodynamic assessment. AB - OBJECTIVE: In this study, a randomized and placebo controlled trial, we aimed to study the effectiveness and safety of doxazosin based upon urodynamic parameters, especially pressure/flow studies, in men with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 57 men (29 doxazosin, 28 placebo) 48-82 years of age with BPH were enrolled. Yet, 8 of 29 in the doxazosin group and 10 of 28 in the placebo group were excluded due to side effects of doxazosin and intolerability of urodynamic assessment of free uroflow, postvoiding residual urine volume (PVR) and pressure/flow studies. RESULTS: There were improvements in all urodynamic parameters (Free Qmax: 30.4% and 28%, PVR: 14 ml and 12 ml, invasive Qmax: 29.3% and 26.2%, Pdet at Qmax: -32.7% and -30%, Pdet-max: -29% and -27.7% at end of the 1st and 6th months whereas placebo effects were worsening in all urodynamic parameters. CONCLUSIONS: We suggest that doxazosin is an important treatment option for patients with BPH, and efficacy of doxazosin should be evaluated with objective, quantitative urodynamic studies not with subjective symptom scores. But additional costs and invasiveness of urodynamic studies restrict their common usefulness. PMID- 10668943 TI - The value of serum prostate specific antigen and other parameters in detecting bone metastases in prostate cancer. AB - The cut-off value of serum prostate-specific antigen (PSA) level in prediction of bone metastases and the correlation of serum PSA with the clinical stage, grade, score and the rate of bone metastases have been investigated in cases of prostate cancer (PCa). The study population consisted of 160 patients with histologically proven PCa between April, 1993 and August, 1996. The negative predictive value and the sensitivity were the highest (94%) in patients with a serum PSA value less than 10 ng/ml. We claim that in patients with PSA values less than 10 ng/ml whole body bone scan is not necessary. PMID- 10668944 TI - Antibiotic prophylaxis for transrectal biopsy of the prostate: a prospective randomized study of the prophylactic use of single dose oral fluoroquinolone versus trimethoprim-sulfamethoxazole. AB - We investigated the efficacy of prophylactic use of single dose oral ofloxacin and trimethoprim-sulfamethoxazole regimens for transrectal prostate biopsy in 110 men. In the ofloxacin, trimethoprim-sulfamethoxazole and control groups, urinary infection was found in 2 (4.76%), 3 (6.66%) and 6 (26.08%) patients, respectively. Both of these antibiotic regimens produced a statistically significant reduction in urinary infection (p<0.02, p<0.05). Our study indicates that single dose fluoroquinolone or trimethoprim-sulfamethoxazole prophylaxis seems to be effective, practical and economical. PMID- 10668945 TI - Prospective evaluation of prostate specific antigen (PSA), PSA density, free-to total PSA ratio and a new formula (prostate malignancy index) for detecting prostate cancer and preventing negative biopsies in patients with normal rectal examinations and intermediate PSA levels. AB - OBJECTIVE: To improve the specificity and sensitivity of prostatic cancer detection, we prospectively evaluated total prostate specific antigen (PSA) level, PSA density, free-to-total PSA ratio and a new formula called prostate malignancy index (PMI) as a discriminator of prostate cancer in patients with intermediate PSA levels and normal digital rectal examinations. MATERIALS AND METHODS: Between November 1995 and October 1997, 95 patients who had serum PSA levels of 4.0 to 10.0 ng/ml with normal digital rectal examinations were prospectively evaluated. All patients underwent one or two times transrectal ultrasound guided prostate biopsies. Based on age specific reference range of PSA, PSA density and % free PSA ratio, PMI was calculated for each patient. The free and total serum PSA concentrations were determined by an Immulite assay system. (Diagnostic Product Corp., Los Angeles, California). RESULTS: Overall 20 of 95 (21%) patients had prostate cancer. There were no significant differences in patient mean age and mean total PSA between those with benign and those with malignant biopsies (p>0.05). However, there were significant differences in mean PSAD, mean free-to-total PSA ratio and mean PMI (p<0.01, p<0.05, p<0.01, respectively). Benign condition specificities for PM index, percent free PSA, PSA density and total PSA at a 90% sensitivity for prostate cancer were 48%, 10.6%, 8% and 4%, respectively. Of 95 patients, 27 (28.4%) had a PMI of equal or more than 3.1, including 12 of 75 (16%) with negative biopsy and 15 of 20 (75%) with positive biopsy. Furthermore a cutoff MI 0.86 P correctly identified 24% of benign cases without missing any prostate cancer cases. The comparison of receiver operating characteristic (ROC) curve areas showed that PMI was better than total PSA (p<0.01). Although, the area under the ROC curve of % free PSA and PSAD were higher than the area of total PSA, these differences were not statistically significant (p>0.05). CONCLUSIONS: We concluded that the prostate malignancy index could be utilized to differentiate benign conditions from prostate cancer in patients with intermediate PSA levels and normal digital rectal examination. Also significant numbers of negative biopsies can be prevented in these patients. PMID- 10668946 TI - Sequential scrotal scintigraphy for the study of varicocele. AB - Scrotal scintigraphy is a non-invasive procedure for evaluating spermatic vein reflux. This technique was used in 43 infertile patients with varicocele. None of them had infections, traumatic, or chromosomal abnormalities that could be associated with their infertility. Twelve patients showed type 1 time activity curve, including 3 with grade II, and 9 with grade III varicocele. A total of 12 patients showed a type 2 pattern and consisted of 2 patients with grade I, 6 with grade II, and 4 with grade III varicocele. Nineteen showed type 3 pattern consisting of 6 with grade I, 6 with grade II, and 7 with grade III. The patients with grade III varicocele showed type 1 pattern more frequently than those with grade I or II (p<0.05). Preoperative sperm concentration in patients with grade I varicocele was significantly lower than that in patients with grade II or III disease (p<0.05, each). Sperm motility of patients with grade I varicocele was also significantly lower than that of those with grade II or III varicocele (p<0.01, each). Patients with grade II or III varicocele showed an increase in sperm concentration postoperatively, and those with grade I varicocele showed a postoperative increase in sperm motility, but the differences between pre- and postoperative values were not significant. While seminal findings in patients with type 2 or 3 pattern did not change after surgery, patients with type 1 pattern showed significant improvement in sperm concentration postoperatively (p<0.05). It is concluded that preoperative sequential scrotal scintigraphy can be a more useful technique for assessing the prognosis for postoperative improvement of seminal findings than the grade decision of varicocele. PMID- 10668947 TI - Significance of serum FSH levels and testicular morphology in infertile males. AB - OBJECTIVES: To determine the relationship between plasma levels of FSH and testicular spermatogenic patterns. METHODS: Testicular biopsies were obtained from 99 infertile men. Biopsies were performed either in order to distinguish the type of azoospermia (obstructive/non-obstructive) or because of severely subnormal semen variables. Serum FSH was measured by immunoassay (normal range is less than 7 mIU/ml). RESULTS: Statistically significant difference was detected between patients with Sertoli cell only syndrome and normal spermatogenesis, hypospermatogenesis and maturation arrest (p<0.01, p<0.01, p<0.05, respectively). No statistically significant differences were found between normal spermatogenesis, hypospermatogenesis and maturation arrest. CONCLUSION: Our study revealed that elevation of serum FSH correlates only with the appearance of Sertoli cell only syndrome. We think that azoospermic or severely oligoasthenoteratozoospermic patients with highly elevated plasma FSH levels (three times the normal) could be excluded from separate testicular biopsy, because these patients are not suitable for conventional treatments. If he is willing to undergo an IVF program the sperm will often be present, no matter what the testicular histology is to be used for assisted reproductive techniques, particularly ICSI. PMID- 10668948 TI - The treatment of penile carcinoma: experience in 64 cases. AB - INTRODUCTION: Carcinoma of the penis is an uncommon entity in Poland (160 new cases per year). PURPOSE: To review our results in treatment of penile cancer in 64 patients. MATERIAL AND METHODS: From 1989 to 1998, 64 patients were treated for carcinoma of the penis. The age of the patients varied from 21 to 86. Clinical and pathological categories were assessed according to TNM classification. Inguinal lymphadenectomy was performed in 35 patients. Following surgery 12 patients underwent radiotherapy, 3 chemotherapy, 3 radiotherapy and chemotherapy. RESULTS: Twenty-two percent of patients died of cancer with median survival of 49 weeks. Bilateral inguinal involvement after node dissection was found in 17 patients. Unilateral inguinal involvement was found in 7 patients. Six patients had positive pelvic nodes. Of patients with initially non metastatic disease (N0) 8.3% showed progression to death, of patients with initially lymph node metastases (N+) 46% showed progression to death. The 5-year disease-free survival rates of patients with N+ and N0 were 40% and 82%, respectively. Of the patients 11% had local recurrence. Postoperative complications developed in 30 cases. CONCLUSIONS: The likelihood of lymph node invasion at presentation was related to T category and grade of primary tumour. The most important prognostic factor for patients with carcinoma of the penis was lymph node involvement. PMID- 10668949 TI - Continuous ambulatory peritoneal dialysis (CAPD) adequacy influences serum free carnitine level. AB - OBJECTIVE: An evaluation of serum free carnitine level in CAPD patients in relation to dietary intake, nutritional status and CAPD adequacy and duration. STUDY DESIGN: Food diaries, nutritional (total body mass, lean body mass, serum level of proteins, carnitine, cholesterol) and adequacy (Kt/V, PCR, tCcr, EN) parameters were obtained in 23 CAPD patients. RESULTS: Normal carnitine level (41.8+/-6.7 micromol/l) was found in 17 patients being on CAPD through 11.1+/-9.6 months, whereas in 6 persons treated with CAPD through 9.7+/-4.1 months carnitine level was 25.4+/-5.7 micromol/l. Significant differences between low and normal carnitine groups were in tCcr (82.7+/-16.7 v. 65.9+/-13.2 l/wk/1.73 m2 BSA), effluent volume (10.9+/-0.8 v. 9.9+/-1.5 l/day), effluent glucose concentration (729+/-167 v. 530+/-220 mg/dl) and serum globulin level (22.6+/-6.4 v. 29.3+/-4.4 g/l). Significant correlation coefficients (for n = 23) were found between serum carnitine level and effluent volume (r=-0.509) or plasma globulin level (r=+0.522). CONCLUSION: Patients with higher CAPD adequacy show lower serum free carnitine levels and this is related to higher effluent volumes. PMID- 10668950 TI - Protein Z levels in haemodialysis patients. AB - Protein Z (PZ) is a vitamin K-dependent protein isolated from human and bovine plasmas. Although the exact role of PZ in the haemostatic system is presently unknown, it is suggested that PZ deficiency may cause bleeding tendency. Haemostatic alterations in end-stage renal failure (ESRF) are certainly complex and involve several abnormalities in the coagulation and fibrinolytic system. In order to elucidate the detail of the haemostasis in ESRF, we aimed to investigate PZ activity in haemodialysis patients. Therefore, we compared plasma PZ levels in 10 haemodialysis patients (6 M, 4 F, mean age 36+/-11) and 10 healthy normal controls (5 M, 5 F, mean age 34+/-8) in this study. We found mean plasma PZ levels in haemodialysis patients and healthy controls 6.95+/-2.93 microg/ml and 3.06+/-0.81 microg/ml, respectively (p<0.005). Increased level of PZ which influences the action of thrombin on its protein substrates and inhibitors may contribute to the haemostatis alterations in ESRF patients, in addition to other well known abnormalities in the coagulation and fibrinolytic system. PMID- 10668951 TI - The importance of bioimpedance (BIA) analysis and Cardio Tens (24-h ABPM and ECG) monitoring in the dialysis programme. AB - The authors performed bioimpedance analysis and Cardio Tens (24-h ABPM and ECG) monitoring in 66 patients (28 males, 38 females) treated in the chronic haemodialysis programme. They investigated the correlations between the body weights before, during and after dialysis, the changes of the water compartments and fat body weight, and the recorded values of blood pressure and ECG alterations. On the basis of the measurements by this non-invasive method it is concluded that, as a result of dialysis and ultrafiltration, the total body weight and total body water are decreasing in a greater extent in men than in women. By gradually decreasing the body weight, the optimal dry weight could be attained, which resulted in the reduction of blood pressure or even normotension. In the course of dialysis the values of bioimpedance and bioreactance increase. The intradialytic hypotensive indispositions were accompanied by a significant reduction of bioreactance (n = 16). The BMI, total body weight and total body water hyperlipidaemic, hypalbuminic patients with treatment-resistant hypertension are considerably larger than those of the patients with normal blood pressure (p<0.01). During Cardio Tens monitoring 53% of the patients proved to be dippers, 47% of whom had ST depression, while in 73% of the non-dippers ischaemic alterations were encountered together with high hyperbaric impact values. The total body weights and total water compartments of patients returning to dialysis with an excess body weight of more than 3.5 kg were significantly larger than of patients who were cooperative and had no oedemas. In the last hour of dialysis and during the following few hours, arrhythmias and ST depressions of the cardiovascularly instable patients appeared more frequently. The total water compartments of these patients are significantly larger than normotensive, normolipaemic patients with appropriate serum albumin concentrations. The importance of the BIA and Cardio Tens monitoring in determining the optimal dry body weight and improving the cardiovascular condition of the patients is emphasized. PMID- 10668952 TI - Tubulointerstitial lesions in IgA nephropathy and localization of hepatocyte growth factor. AB - To investigate the relationship between localization of hepatocyte growth factor (HGF) and tubulointerstitial lesions (TILs) in the cortical area of renal biopsy specimens, a clinicopathological study was performed in 55 patients with IgA nephropathy. HGF was detected by an enzyme-antibody method and TILs were assessed semiquantitatively by light microscopy. HGF was observed mainly on epithelial cells in the tubules, but not in the glomeruli. Fourteen patients had biopsies that were positive for HGF. There was a correlation between HGF positivity and histological damage, the TIL grade, and several clinical parameters determined at biopsy. Thus, HGF is related to TILs in IgA nephropathy and may be a factor in the exacerbation of this disease. PMID- 10668953 TI - Relationship between some prognostic markers of HD patients and serum erythropoietin, insulin-like growth factor-1, leptin, parathormone and testosterone. AB - Iseki et al. [1] have shown that serum levels of albumin (Alb), creatinine (Cr) and BMI are significant predictors of death in haemodialyzed patients (HD pts). In our study we decided to assess the relationship between the levels of Alb, Cr, BMI and substances which have a known metabolic effect on nutritional status in HD pts: endogenous erythropoietin (Epo), insulin-like growth factor-1 (IGF-1), leptin (Lep), parathormone (PTH), and testosterone. The study was conducted in 53 (28M, 25F) stable HD pts. Serum levels of endogenous Epo and PTH were estimated by CLIA; IGF-1, Lep, testosterone, sex hormone binding globulin were estimated by RIA. The multiple regression analysis was done between Alb, Cr, BMI and Epo, IGF 1, PTH and Lep for all HD pts together and free androgen index (FAI) for men and women separately. Correlations: the level of serum albumin did not correlate significantly with any of the measured substances. Serum creatinine level significantly correlated only with the level of IGF-1 (p=0.02), BMI was significantly correlated with serum endogenous Epo (p<0.01), leptin (p=0.004) and FAI (p<0.005) both in men and women. We concluded that the higher concentrations of endogenous Epo, IGF-1 and testosterone could be correlated with a better prognosis in HD patients. PMID- 10668954 TI - Tuberculosis in chronic renal failure in Jeddah. AB - The incidence of tuberculosis is still high in many developing countries and immunocompromised patients with chronic renal failure requiring haemodialysis have been reported to be at increased risk of developing tuberculosis. In this study 80 patients with chronic renal failure were followed up for a period of three years and were carefully monitored for the development of tuberculosis. Mantoux test, chest radiograph and sputum were performed at the beginning of the study and every six months thereafter. At the end of the study period, 8 (10%) of the patients had developed tuberculosis, confirming the high incidence of tuberculosis in this group of patients. No particular underlying renal disease was associated with the development of tuberculosis. Four patients developed pulmonary tuberculosis, 2 renal tuberculosis and one each with cervical tuberculous lymphadenitis and tuberculous meningitis. All patients responded satisfactorily to anti-tuberculosis therapy as diagnosis was established early. The delayed recognition of tuberculosis and therefore a delay in the initiation of effective treatment is not only detrimental to the patient but also results in a potentially profound impact on public health. We recommend routine screening for tuberculosis in patients with chronic renal failure presenting at Renal Units and tuberculosis chemoprophylaxis for those undergoing haemodialysis, particularly in countries with high incidence of tuberculosis. PMID- 10668955 TI - Third single chromosome 6 workshop: meeting report. PMID- 10668956 TI - The chromosome 6 sequencing project at the Sanger Centre. AB - Chromosome 6 is probably best known for encoding the major histocompatibility complex (MHC) which is essential to the human immune response. In addition, it has been shown to be associated with many diseases such Schizophrenia, Diabetes, Arthritis, Haemochromatosis, Narcolepsy, Epilepsy, Retinitis Pigmentosa, Deafness, Ovarian Cancer, and many more. Chromosome 6 is about 180 Mb in size and is estimated to encode around 3500 genes of which only about 10% are currently known. It is our aim to map, sequence and annotate the entire chromosome in close collaboration with the chromosome 6 community. PMID- 10668957 TI - Retroelements and segmental duplications in the generation of diversity within the MHC. PMID- 10668958 TI - A linkage map of chromosome 6 based on 2 large kindreds segregating bipolar affective disorder. AB - Twenty-eight markers, both simple sequence repeats (SSRs) and restriction fragment length polymorphisms (RFLPs), were genotyped on members of 2 large pedigrees (OOA, BIP167) segregating bipolar affective disorder. Using the multipoint program "build" of CRIMAP and odds of placement 1000:1, a unique sex averaged map was generated that spans 227 cM and includes 26 markers. The two other markers were placed on the map with a lower likelihood. The female recombination map is larger than the male recombination map by about 80%. Linkage analysis between the polymorphisms and the disease in the OOA screening pedigree did not result in any significantly positive lod scores nor did a non-parametric, identity-by-descent, method generate any significant p-values. BIP167 was analyzed for allele sharing at the simple sequence repeat loci and significant associations were not found. At present we conclude, that the pedigrees under study do not have a major predisposition gene for bipolar affective disorder on chromosome 6 under the diagnostic and transmission models analyzed by the 2 different methods. PMID- 10668959 TI - Isolation and characterisation of cosmids to intervals within a 4.5Mb region at 6p21.3. AB - The gene responsible for hereditary haemochromatosis (HH) has recently been identified. One mutation in this gene, termed HFE, has been found in all Australian HH patients. We previously identified a predominant HH ancestral haplotype covering 4.5Mb at 6p21.3, and showed that patients with two copies of this haplotype express a more severe form of the disorder. One key question to now be resolved is why haplotype related variation in phenotypic expression of HH is present if all patients tested have the same HFE mutation. A cosmid resource covering the 4.5Mb HH ancestral haplo type region was obtained. These cosmids provide the material for the completion of a transcript map of this region, and will assist the identification of candidate modifiers of HFE expression. PMID- 10668961 TI - Alternative splicing of the LST-1 gene located in the Major Histocompatibility Complex on human chromosome 6. PMID- 10668960 TI - From long range mapping to sequence-ready contigs on human chromosome 6. AB - Our aim is to construct physical clone maps covering those regions of chromosome 6 that are not currently extensively mapped, and use these to determine the DNA sequence of the whole chromosome. The strategy we are following involves establishing a high density framework map of the order of 15 markers per Megabase using radiation hybrid (RH) mapping. The markers are then used to identify large insert genomic bacterial clones covering the chromosome, which are assembled into sequence-ready contigs by restriction enzyme fingerprinting and sequence tagged site (STS) content analysis. Contig gap closure is performed by walking experiments using STSs developed from the end sequences of the clone inserts. PMID- 10668962 TI - Complex duplications at 6p22.1, 6p11.2, 5q13, 5p15.1 and 5p13 revealed by fluorescent in situ hybridisation. AB - A complex pattern of fluorescent in situ hybridisation (FISH) has been detected using PAC clones from the short arm of chromosome 6, proximal to the haemochromatosis gene at 6p22.1. Cross-hybridisation to 6p22.1, 6p11.2, 5q13, 5p15.1 and 5p13 was consistently detected with several PAC clones covering a genomic region greater than 200 kb. These results indicate that large sections of genomic DNA are shared by these 5 disparate chromosomal segments, indicative of large scale duplication events. These results were in part accounted for by the identification of several expressed sequence tags (ESTs). PMID- 10668964 TI - Physical mapping of two histone gene clusters on human chromosome 6p22.1-22.2. AB - Histones are basic proteins which are responsible for the assembly and maintenance of the nucleosomal structure within the chromosomal fiber in eukaryotes. Two clusters of these genes have previously been mapped to the region 6p21.1-p22.2. We describe here a radiation hybrid map, a long range restriction map and a YAC contig covering and linking these two clusters and giving the precise localisation with respect to the HLA complex. The large cluster contains five H1 histone genes in the 6p22.2 region, the smaller only one, H1F5 (H1.5), in 6p22.1. In both clusters, each H1 locus is accompanied by several core histone genes. The large cluster has additionally been covered by a sequence ready PAC contig and three probably unrelated genes (TRMI2, BTN and SSADH) have been accurately localized within the 6p22.2-p22.1 region. PMID- 10668963 TI - The Chromosome 6 database at the Sanger Centre. AB - The Sanger Centre Chromosome 6 Database (6ace) has been developed as the primary means of release of annotated sequencing and mapping information for human chromosome 6 from the Sanger Centre. It is also being used to curate global data from published and unpublished external sources. The rationale behind the development of 6ace is described, together with information as to how to access the database. PMID- 10668965 TI - Dissection of the 5.5 Mbp region directly telomeric of HLA-B including a long range restriction map, YAC and PAC contigs. AB - A large number of diseases are associated with the human major histocompatibility (HLA) complex located in 6p21.3. The underlying defect of most of these has not yet been determined even after detailed analysis of the HLA region. Due to the extended haplotypes found in this area, several of the HLA-linked disease genes may be located also telomeric of the class I region. In order to analyse the area covering the 4 megabases directly telomeric of HLA-F in close detail, we have generated 50 new markers. These and other markers have been used to establish a SalI restriction map from 46 YACs. A subset of 42 markers was applied to construct a genomic long range restriction map from an HLA-A2/B13 haplotype. Both maps have been compared revealing the presence of additional 150 kb in the HLA-A2 haplotype close to the RFP locus. Additionally, 47 PACs have been selected mapping to this region and grouped into 7 contigs. Sequencing of these PAC contigs has already been initiated. PMID- 10668966 TI - Biology of chromosome 6. AB - The major histocompatibility (HLA) complex on the short arm of human chromosome 6 has attracted many scientists over the last three decades. It is the purpose of this brief review to point out that the remaining large regions of chromosome 6 contain genes involved in several interesting biological phenomena as well. Focus will be particularly on genes affecting behavioural features and sensory perception. The likely involvement of HLA and closely linked olfactory receptor loci in mate selection and their possible role in favouring heterozygosity among the offspring will also be discussed. PMID- 10668967 TI - A brief history of the Hubrecht Laboratory. PMID- 10668968 TI - A treasure house of comparative embryology. AB - The Embryo Collection of the Hubrecht Laboratory is a treasure house of comparative embryology. It is the largest and most important collection of its kind in the world, and consists of thousands of vertebrate embryos stored in alcohol, or prepared as histological sections. Many elusive species are included in the collection, some represented by complete developmental series. The accompanying archives offer a remarkable insight into the methods used to collect embryos form wild animals, as well as the motives behind the founders of the collection. Carefully maintained, documented and catalogued, the collection is available for study by all interested scientists. We argue that this collection is one of the greatest biodiversity resources in existence. PMID- 10668969 TI - A nose for the embryo: the work of Pieter Nieuwkoop. PMID- 10668970 TI - Pieter Nieuwkoop's contributions to the understanding of meso-endoderm induction and neural induction in chordate development. AB - Pieter Nieuwkoop, who died September 18, 1996, at age 79 in Utrecht, The Netherlands, is remembered by developmental biologists for his numerous research contributions and integrative hypotheses over the past 50 years, especially in the areas of neural induction, meso-endoderm induction, and germ cell induction in chordates. Most of his experimentation was done on the embryos of amphibia, the preferred vertebrate embryo of the early years of the 20th century. One of his last publications contains a comparison of the experimental advantages and disadvantages of anuran and urodele amphibians (Nieuwkoop, 1996). The significance of his findings and interpretations for developmental biology can be estimated from the fact that researchers of many laboratories worldwide continue to work on the phenomena he first described and to extend the hypotheses he first formulated. The aim of this article is to review Nieuwkoop's main contributions and to cite the recent extensions by others. PMID- 10668971 TI - The neural induction process; its morphogenetic aspects. AB - This posthumous review of early embryonic inductions concludes: 1) the amphibian egg has only two distinct components, animal and vegetal. Interactions at their mutual boundary forms meso-endoderm. This is "meso-endoderm induction", not just "mesoderm induction". 2) The dorso-ventral polarity of the yolk mass implies a dorsally situated inducing centre. 3) Accumulation of cells into one, two, three or many cell masses [problastopores] along the circumference of the meso-endoderm results in as many axes, implying a self-organizing capacity of meso-endoderm. 4) Induction of the meso-endoderm is slow, spreading cell to cell through the animal moiety from the boundary of the vegetal yolk mass towards the animal pole. 5) Interaction between mesoderm and ectoderm is a separate step leading to cranio caudal differentiation of the archenteron roof. 6) The initial invaginating endoderm and mesoderm, representing the future pharynx endoderm and prechordal plate mesoderm, first contacts the most posterior presumptive neurectoderm after having passed the still uninvaginated trunk mesoderm. At that moment an antero posterior level neural induction actually starts. 7) The ectoderm contraction wave coincides spatially and temporally with the induced neural plate. 8) Two successive homoiogenetic waves of inductive activity pass through the presumptive neurectoderm in the anterior direction, the first one, "activation", giving rise to neural differentiation and ultimately forebrain, the second one, "transformation", to more caudal CNS structures. These are separate, successive steps in CNS regional induction. 9) The midbrain represents a secondary formation in the neural plate. 10) The observed changes in morphogenesis may depend upon separate, successive binary decisions via [cell and] nuclear state splitters [involving differentiation waves]. PMID- 10668972 TI - Discovery of a morphogen. PMID- 10668973 TI - A visit to the Hubrecht laboratory. PMID- 10668974 TI - Initiation, establishment and maintenance of Hox gene expression patterns in the mouse. AB - Spatially and temporally restricted expression of the Hox genes along the main and appendicular axes is essential for correct patterning of vertebrate embryos. In this overview we discuss the latest data that shed light on the mechanisms underlying the generation of the expression domains of the Hox genes. The molecular genetic interactions governing initial transcription of the Hox genes in the posterior part of the primitive streak during mouse and chick gastrulation remain enigmatic. But the recent discovery by Kondo and Duboule (Cell, 97, 1999, 407-417) of a "cluster repressive regulation", will undoubtedly lead to a better understanding of the molecular genetic mechanism underlying colinear and sequential initiation of Hox gene transcription. Recently progress has been booked in characterizing the basal processes driving progression of the Hox expression domains during their establishment. Hox expression is still labile while being established. The transcriptional state of Hox genes in anterior tissues can be reprogrammed under the influence of more posterior locations. Posteriorizing activity may involve RA and FGF signaling. It is only when these interactions and, in some cases at least, regulatory interactions with Hox and cdx gene products occur appropriately, that the Hox expression domains would be correctly established. After the Hox expression domains have been established, regulatory processes involving the products of Polycomb and trithorax- Group genes start operating, perpetuating the transcriptional state of the Hox genes within and outside the expression domains. Whether control at the level of chromatin structure, believed to operate during the late maintenance phase of Hox gene expression, is also involved in regulating concerted initial expression of these genes, is a possibility that has been suggested. PMID- 10668975 TI - Vertebrate aristaless-related genes. AB - Aristaless-related genes, a subset of the Paired-related homeobox genes, have in the past few years emerged as a group of regulators of essential events during vertebrate embryogenesis. One group of aristaless-related genes has been linked to the morphogenesis of the craniofacial and appendicular skeleton by their expression patterns and by the phenotypes of natural and artificial mouse mutants. Expression and function in the nervous system characterise a second group, and a third group, the Pitx genes, have been shown to have many different roles, including functions in the pituitary, left-right determination and limb development. PMID- 10668976 TI - Novel interactions between vertebrate Hox genes. AB - Understanding why metazoan Hox/HOM-C genes are expressed in spatiotemporal sequences showing colinearity with their genomic sequence is a central challenge in developmental biology. Here, we studied the consequences of ectopically expressing Hox genes to investigate whether Hox-Hox interactions might help to order gene expression during very early vertebrate embryogenesis. Our study revealed conserved autoregulatory loops for the Hox4 and Hox7 paralogue groups, detected following ectopic expression Hoxb-4 or HOXD4, and Hoxa-7, respectively. We also detected specific induction of 5' posterior Hox genes; Hoxb-5 to Hoxb-9, following ectopic expression of Hoxb-4/HOXD4; Hoxb-8 and Hoxb-9 following ectopic expression of Hoxa-7. Additionally, we observed specific repression of 3' anterior genes, following ectopic expression of Hox4 and Hox7 paralogues. We found that induction of Hoxb-4 and Hoxb-5 by Hoxb-4 can be direct, whereas induction of Hoxb-7 is indirect, suggesting the possibility of an activating cascade. Finally, we found that activation of Hoxb-4 itself and of posterior Hox genes by Hoxb-4 can be both non-cell-autonomous, as well as direct. We believe that our findings could be important for understanding how a highly ordered Hox expression sequence is set up in the early vertebrate embryo. PMID- 10668977 TI - A tight control over Wnt action. AB - Here, we review the WNT pathway and its regulation at different levels. We focus on the transcriptional regulation of WNT target genes, in light of the recently identified negative regulators, i.e. relatives of groucho and CBP. PMID- 10668978 TI - Growth factor signalling. AB - Signalling between cells in the developing vertebrate embryo is essential for normal embryonic development. In the mid 1970's, signal transduction research started at the Hubrecht Laboratory with special emphasis on analysis of the signalling mechanisms that direct cell proliferation and differentiation. The introduction of in vitro model systems contributed tremendously to the success of the signal transduction research at the Hubrecht Laboratory. Initially neuroblastoma cell lines, and later embryonal carcinoma and embryonal stem cells played an important role in identification of the molecular key players in developmental signalling. For instance, embryonal carcinoma cells were used to identify and characterise polypeptide growth factors. Growth factor signalling research was extended to analysis of growth factor receptor activation. Moreover, the second messenger systems that are linked to growth factor receptors were studied, as well as the nuclear responses to growth factor receptor activation. Finally, the role of growth factor signalling in differentiation was established using embryonal carcinoma cells. Here, we will review work that was characteristic for the growth factor receptor signalling research that was done at the Hubrecht Laboratory between 1980 and the early 1990's. PMID- 10668979 TI - Developmental tumours, early differentiation and the transforming growth factor beta superfamily. AB - Embryonal carcinoma and embryonic stem cells have been very useful models for identifying some of the factors that regulate differentiation in early mammalian development. Here, we present a brief history of their original isolation and characterization and of their later introduction into the Hubrecht Laboratory. We illustrate in a review their contribution to our current understanding of the function of transforming growth factor beta and ligands binding to the receptors of a related factor, activin, in development with some of our own work. PMID- 10668980 TI - Signals governing extraembryonic endoderm formation in the mouse: involvement of the type 1 parathyroid hormone-related peptide (PTHrP) receptor, p21Ras and cell adhesion molecules. AB - The formation of parietal endoderm (PE) from primitive endoderm (PrE) immediately after implantation of the early mouse embryo can be seen as the earliest example of an epithelio-mesenchyme transition (EMT) in murine development. Since EMT and EMI (epithelium-mesenchyme interactions) are at the very heart of morphogenesis, identifying molecular mechanisms governing these processes is of utmost importance. An excellent in vitro model system to study PE formation, i.e. F9 embryonal carcinoma cells, is available to this end. In the present paper we review our own recent results and those of others using these cells, and present our current view on the molecular mechanisms involved in PE formation. PMID- 10668981 TI - Receptor protein-tyrosine phosphatase signalling in development. AB - Receptor Protein-Tyrosine Phosphatases (RPTPs) belong to the superfamily of protein-tyrosine phosphatases and have the intrinsic ability to transduce signals across the cell membrane. We are beginning to understand the role of RPTPs in development of invertebrates, due to elegant genetic studies. In contrast, relatively little is known about the role of RPTPs in vertebrate development. Signalling by RPTPs has predominantly been studied in mammalian cell systems, which has led to important insights into potential ligands, into regulation of RPTP activity and into potential RPTP substrates. Here, we will introduce the RPTPs, and discuss the function of the LAR-subfamily of RPTPs. In addition, we focus on the function and signalling of the haematopoietic RPTP, CD45. Finally, we will discuss the structure and function of RPTPalpha, the RPTP that is the subject of our studies. PMID- 10668982 TI - Morphogenetic action of retinoids and estrogens. AB - Retinoids and estrogens are small lipophilic compounds fulfilling important biological roles in vertebrate development, reproduction and homeostasis. Both types of ligands are regulators of gene transcription by binding to (nuclear) proteins acting as ligand-activatable transcription factors, members of the nuclear receptor gene superfamily. Retinoids and their multiple receptors (RARs/RXRs) are particularly well-known for their role in early development and spermatogenesis, while much less is known about the two estrogen receptors (ERalpha/beta) during development. In this article we describe some of our previous and present work in both areas of research. PMID- 10668983 TI - Germ line development in fishes. AB - Classical work on germ cells in fishes has dealt with three main issues; their embryonic origin, the proliferation, and migration pathway during embryonic and larval development. Until recently, primordial germ cells (PGCs) have been studied in a number of fishes using morphological criteria only. The identification of the Drosophila vasa homolog gene of zebrafish now allows comparison of these morphological data with vasa RNA expression patterns in zebrafish. Teleost PGCs can be distinguished from somatic cells by their distinct morphology, at the earliest during gastrulation, and in most fishes their number varies between 10 and 30 during pregonial development. Mitosis is generally not observed in PGCs at extragonadal locations, whereas they are mitotically active once at the gonadal ridges. During gastrulation, PGCs appear to translocate from the epiblast to the hypoblast and during somitogenesis they are found associated with the most peripheral yolk syncitial layer (YSL). From the peripheral YSL they migrate through the median mesoderm into the dorsal mesoderm and then to the dorsal mesentery, where they establish the gonad primordia with mesenchymal cells. Vasa RNA positive cells, the PGCs of the zebrafish conform to these general observations. Interestingly, classical descriptive and experimental data can now be reevaluated using vasa as a molecular marker of the fish germ line. The power of zebrafish genetics together with possibilities of experimental embryology should accelerate research on aspects of vertebrate germ line development such as PGC migration, division and apoptosis, as well as (in) fertility. The present review summarizes some of the classical data on germ line development in fishes in relation to recent data on vasa expression in zebrafish and compares these findings, where appropriate, with those in other model organisms. Special emphasis is placed on vasa gene expression as a potential universal germ line marker and suggestions are made for novel, zebrafish specific approaches to investigate the vertebrate germ line. PMID- 10668984 TI - Developmental bioinformatics: linking genetic data to virtual embryos. AB - This paper discusses current efforts to produce databases of gene expression for the major model embryos used in developmental biology. The efforts to build these resources were motivated by the need for immediate internet access to all types of research data, and the production of these databases is a major and new challenge for bioinformatics. Thus far bioinformatics has mainly been concerned with textually oriented resources and data, much of it concerned with gene and protein sequences. Because the genetic basis of developmental biology is integrated with developmental anatomy, these databases require the use of images to link molecular data with spatial information. In order to standardise database formats, digital atlases of some model systems are being produced that include integrated anatomical descriptions and these are being linked to appropriate genetic data. Integrating such image-based, searchable data into databases makes new demands on the field of bioinformatics and we consider here the imaging modalities that are used to obtain information and we discuss in particular the production of 3D images from serial sections. Next, we consider how to integrate textual and spatial descriptions of gene expression and the key tool needed to make this possible, i.e. anatomical nomenclature. A short review of internet resources on developmental biology is also given and future prospects for the development of these databases are discussed. PMID- 10668985 TI - Fate mapping the mouse embryo. AB - The use of clonal analysis to obtain a fate map of the epiblast of the mouse embryo and to investigate cell distribution during gastrulation and early neurulation is described in a personal reminiscence. A revised fate map of the epiblast at 6.5 days gestation is provided, and the development of 3-dimensional, quantitative image analysis techniques outlined. PMID- 10668986 TI - Comparative and correlative neuroanatomy for the toxicologic pathologist. AB - Xenobiotic-induced neuroanatomic alterations are always regarded as adverse and are commonly used to define reference doses to manage neurotoxic risk. Thus, the neuropathologist plays an essential role in evaluating potential neurotoxicants. The pathologist must be able to recognize the morphologic differences that exist among species, strains, and ages or between genders (comparative neuroanatomy) and to grasp the impact of structural damage on neural function (correlative neuroanatomy). Brain anatomy and function may be used to group the mammals used in neurotoxicity bioassays into 3 classes: rodent, carnivore, and primate. Neural function may or may not be affected by the structural divergence. Rodents are preferred for neurotoxicity assays because their reduced body size allows optimal perfusion at little cost and their smaller brain size permits screening of multiple regions using few sections. However, care must be exercised when interpreting rodent neuropathology data because the rodent paleocortex does not recapitulate the sophisticated neocortical circuitry and functions of carnivores and primates. Knowledge of the neuroanatomic variations that exist among test species assists the neuropathologist in defining the relevance of structural alterations, the potential clinical sequelae of such findings, and the possible significance of similar changes in humans. PMID- 10668987 TI - Microglial response to brain injury: a brief synopsis. AB - In addition to astrocytes and oligodendrocytes, microglia represent the third major population of glial cells within the central nervous system (CNS). Microglia are distributed ubiquitously throughout the brain and spinal cord, and one of their main functions is to monitor and sustain neuronal health. Microglial cells are quite sensitive to even minor disturbances in CNS homeostasis, and they become readily activated during most neuropathologic conditions, including peripheral nerve injury, trauma and stroke, inflammatory disease, and neurotoxicant-induced neuronal injury. During activation, microglia display conspicuous functional plasticity, which involves changes in cell morphology, cell number, cell surface receptor expression, and production of growth factors and cytokines. The many changes occurring in activated cells reflect the altered functional states of microglia that are induced by signals arising from injured neurons. Thus, neuronal-microglial signaling plays a fundamental role in understanding how the CNS responds to injury. Reactive microgliosis should be viewed as a cellular effort to initiate ameliorative and reparative measures in the injured brain. PMID- 10668988 TI - Synaptic clefts are made to be crossed: neurotransmitter signaling in the central nervous system. AB - The primary means of communication between neurons in the mammalian central nervous system (CNS) is via release of chemical transmitters. Although the first transmitters to be discovered were the biogenic amines, such as acetylcholine and norepinephrine, involved in transmission in the autonomic nervous system, the contribution of other types of transmitters, such as amino acids and peptides, to CNS transmission has been the subject of recent study. Part of this interest stems from the relatively large percentage of neuronal connections that make use of amino acid transmitters such as gamma-aminobutyric acid and glutamate and also from the tremendous diversity possible when peptides are used as transmitters. Several disorders of CNS transmission are related to the degeneration of neuronal pathways in the brain. Two of the most prevalent neurologic disorders that result from degeneration are Alzheimer's and Parkinson's diseases. Aspects of these disorders related to chemical neurotransmission are discussed, along with implications with regard to therapeutic strategies. Functions of and possible abnormalities in amino acid transmission that may be associated with CNS disorders are examined. Several peptides are postulated to play a role in neurotransmission and concepts regarding the significance of the coexistence and release of biogenic amines and peptides at the same neuronal terminals are presented. PMID- 10668989 TI - An integrative approach to neurotoxicology. AB - Exposure of human populations to a wide variety of chemicals has generated concern about the potential neurotoxicity of new and existing chemicals. Experimental studies conducted in laboratory animals remain critical to the study of neurotoxicity. An integrative approach using pharmacokinetic, neuropathological, neurochemical, electrophysiological, and behavioral methods is needed to determine whether a chemical is neurotoxic. There are a number of factors that can affect the outcome of a neurotoxicity study, including the choice of animal species, dose and dosage regimen, route of administration, and the intrinsic sensitivity of the nervous system to the test chemical. The neurotoxicity of a chemical can vary at different stages of brain development and maturity. Evidence of neurotoxicity may be highly subjective and species specific and can be complicated by the presence of systemic disease. The aim of this paper is to give an overview of these and other factors involved in the assessment of the neurotoxic potential for chemicals. This article discusses the neurotoxicity of several neurotoxicants (eg, acrylamide, trimethyltin, 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine, manganese, and ivermectin), thereby highlighting a multidisciplinary approach to the assessment of chemically induced neurotoxicity in animals. These model chemicals produce a broad range of effects that includes peripheral axonopathy, selective neuronal damage within the nervous system, and impaired neuronal-glial metabolism. PMID- 10668990 TI - Mechanisms of injury in the central nervous system. AB - Neurotoxicants with similar structural features or common mechanisms of chemical action frequently produce widely divergent neuropathologic outcomes. Methylmercury (MeHg) produces marked cerebellar dysmorphogenesis during critical periods of development. The pathologic picture is characterized by complete architectural disruption of neuronal elements within the cerebellum. MeHg binds strongly to protein and soluble sulphydryl groups. Binding to microtubular -SH groups results in catastrophic depolymerization of immature tyrosinated microtubules. However, more mature acetylated microtubules are resistant to MeHg induced depolymerization. In contrast to MeHg, the structurally similar organotin trimethyltin (TMT) elicits specific apoptotic destruction of pyramidal neurons in the CA3 region of the hippocampus and in other limbic structures. Expression of the phylogenetically conserved protein stannin is required for development of TMT induced lesions. Inhibition of expression using antisense oligonucleotides against stannin protects neurons from the effects of TMT, suggesting that this protein is required for expression of neurotoxicity. However, expression of stannin alone is insufficient for induction of apoptotic pathways in neuronal populations. The aromatic nitrocompound 1,3-dinitrobenzene (DNB) has 2 independent nitro groups that can redox cycle in the presence of molecular oxygen. Despite its ability to deplete neural glutathione stores, DNB produces edematous gliovascular lesions in the brain stem of rats. Glial cells are susceptible despite high concentrations of reduced glutathione compared with neuronal somata in the central nervous system (CNS). The severity of lesions produced by DNB is modulated by the activity of neurons in the affected pathways. The inherent discrepancy between susceptibility of neuronal and glial cell populations is likely mediated by differential control of the mitochondrial permeability transition in astrocytes and neurons. Lessons learned in the mechanistic investigation of neurotoxicants suggest caution in the evaluation and interpretation of structure-activity relationships, eg, TMT, MeHg, and DNB all induce oxidative stress, whereas TMT and triethyltin produce neuronal damage and myelin edema, respectively. The precise CNS molecular targets of cell-specific lipophilic neurotoxicants remain to be determined. PMID- 10668991 TI - Mechanisms of toxic injury in the peripheral nervous system: neuropathologic considerations. AB - The anatomical distribution and organization of the peripheral nervous system as well as its frequent ability to reflect neurotoxic injury make it useful for the study of nerve fiber and ganglionic lesions. Contemporary neuropathologic techniques provide sections with excellent light-microscopic resolution for use in making such assessments. The histopathologist examining such peripheral nerve samples may see several patterns of neurotoxic injury. Most common are axonopathies, conditions in which axonal alterations are noted; these axonopathies often progress toward the Wallerian-like degeneration of affected fibers. These are usually more severe in distal regions of the neurite, and they affect both peripheral and central fibers. Examples of such distal axonopathies are organophosphorous ester-induced delayed neuropathy, hexacarbon neuropathy, and p-bromophenylacetylurea intoxication. These axonopathies may have varying pathologic features and sometimes have incompletely understood toxic mechanisms. In such neuropathies with fiber degeneration, peripheral nerve axons may regenerate, which can complicate pathologic interpretation of neurotoxicity. On occasion neurotoxins elicit more severe injury in proximal regions of the fiber (not included in this review). Axonal pathology is also a feature of the neuronopathies, toxic states in which the primary injuries are found in neuronal cell bodies. This is exemplified by pyridoxine neurotoxicity, where there is sublethal or lethal damage to larger cytons in the sensory ganglia, with failure of such neurons to maintain their axons. Lastly, one may encounter myelinopathies, conditions in which the toxic effect is on the myelin-forming cell or sheath. An example of this is tellurium intoxication, where demyelination noted in young animals is coincident with toxin-induced interference of cholesterol synthesis by Schwann cells. In this paper, the above-noted examples of toxic neuropathy are discussed, with emphasis on mechanistic and morphologic considerations. PMID- 10668992 TI - Application of silver degeneration stains for neurotoxicity testing. AB - Silver staining procedures have been used in numerous ways to render a variety of physical and biological features visible. In biological tissue, histologic protocols use silver to visualize diverse structures or features, such as reticulin, melanin, fungi, chromosome bands, nucleolar organizing regions, and different features in the nervous system. A comparison of the specific steps in these protocols indicates that the silver is "directed" to stain any given feature by the type of fixation, the pretreatment ("mordanting"), the composition of the silver-containing solution(s), and the form of development (reduction). Since the mechanisms of staining have not been understood historically (nor are they now), each method was developed by trial and error. Keystone methods such as those of Bodian and Bielschowsky exploit the nervous system's affinity for silver (argyrophilia). The beginning of a new era in brain research came with the recognition that distinct silver-impregnated morphologic changes occurring in damaged axons could be used for tracing axon pathways in experimental animals with specifically placed lesions. Improvements in staining methods used to selectively impregnate the disintegrating axons but to leave normal axons unstained were achieved by Nauta and Gygax (early workers with these procedures) and spawned a host of method variations known as the "Nauta" methods. Of these, the Fink-Heimer and de Olmos cupric-silver methods were able to unambiguously demonstrate disintegrating synaptic terminals, thereby allowing complete tracing of axon pathways. The late 1970s and 1980s witnessed innovative applications of these techniques. The silver methods once used to trace axon pathways became indicators of the extreme endpoint of neurotoxicity: disintegrative degeneration of neurons induced by neurotoxic chemicals that were administered systemically. The hallmark of neurotoxic substances is the selectivity with which each destroys specific populations or subpopulations of neurons. The high contrast and sensitivity of the silver degeneration stains greatly facilitate the screening process to detect these affected populations, especially when there is no basis for knowing where in the brain to look for damage. More recently, in addition to expanded use in screening for neurotoxic effects, the silver degeneration stains are being used to chart the neuron populations undergoing programmed cell death in the developing brain. Other newly developed silver methods have been refined to show nondisintegrative degeneration, such as the plaques,and tangles of Alzheimer's disease. PMID- 10668993 TI - MK-801 neurotoxicity in cupric silver-stained sections: lesion reconstruction by 3-dimensional computer image analysis. AB - Routine histopathologic evaluation of the brain (paraffin embedding, hematoxylin and eosin staining) makes it difficult for an investigator to identify the overall location and relative extent of lesions as they relate to neural substructures. Moreover, it is very difficult to convey this information to others who are less familiar with neuroanatomy. This study combined a 3 dimensional imaging program with a cupric silver stain for neuronal degeneration in order to determine the location and extent of a focal lesion produced by MK 801 (dizocilpine maleate), a glutamate receptor antagonist that induces necrosis in a small population of neurons in the cortex of rats. A male Sprague-Dawley rat was treated with a subcutaneous dose of MK-801 (10 mg/kg) and was perfused with fixative through the left ventricle 3 days after treatment, a time point known to reveal maximal neurotoxic effects. The brain was embedded in a gelatin matrix, frozen, and serially sectioned at a thickness of 40 microm. The cupric silver method of de Olmos was used to stain frozen sections at 320-microm intervals. Using a color charged-couple device (CCD) camera and a macro lens, a series of 2 dimensional images, which encompassed the entire rostral to caudal extent of the brain, was captured. A computer program was written to define internal and external boundaries in these 2-dimensional images. Then, 3-dimensional reconstructions were generated on a Silicon Graphics workstation using IRIS "Explorer." The quality of the 3-dimensional reconstructions allowed for easy identification of various neural substructures while clearly revealing the exact location and extent of the resulting necrotic neurons that were positively identified by the cupric silver stain. This 3-dimensional lesion reconstruction method provides a powerful tool for conveying spatial information about the nature of neurotoxic lesions in the brain. In addition, it may be used to investigate further dose-response relationships and the effects of other neurotoxicants. PMID- 10668994 TI - Fluoro-Jade: novel fluorochromes for detecting toxicant-induced neuronal degeneration. AB - Two anionic fluorescein derivatives can be used for the simple and definitive localization of neuronal degeneration in brain tissue sections. Initial work on the first generation fluorochrome, Fluoro-Jade, demonstrated the utility of this compound for the detection of neuronal degeneration induced by a variety of well characterized neurotoxicants, including kainic acid, 3-nitropropionic acid, isoniazid, ibogaine, domoic acid, and dizocilpine maleate (MK-801). After validation, the tracer was used to reveal previously unreported sites of neuronal degeneration associated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), methamphetamine, and d-fenfluramine. Preliminary findings with a second generation fluorescein derivative, Fluoro-Jade B, suggest that this tracer results in staining of optimal contrast and resolution in animals dosed with kainic acid. These 2 tracers can be combined with other histologic methods, including immunofluoresence and fluorescent Nissl stains. Recent preliminary findings on a number of specialized applications of Fluoro-Jade include the detection of apoptosis, amyloid plaques, astrocytes, and dead cells in tissue culture. PMID- 10668995 TI - Virtual neuropathology: three-dimensional visualization of lesions due to toxic insult. AB - A first-pass approach incorporating high-field magnetic resonance imaging (MRI) was used for rapid detection of neuropathologic lesions in fixed rat brains. This inherently 3-dimensional and nondestructive technique provides high-resolution, high-contrast images of fixed neuronal tissue in the absence of sectioning or staining. This technique, magnetic resonance microscopy (MRM), was used to identify diverse lesions in 2 well-established rat neurotoxicity models. The intrinsic contrast in the images delineated lesions that were identified using a battery of histologic stains, some of which would not be used in routine screening. Furthermore, the MRM images provided the locations of lesions, which were verified upon subsequent sectioning and staining of the same samples. The inherent contrast generated by water properties is exploited in MRM by choosing suitable pulse sequences, or proton stains. This approach provides the potential for a comprehensive initial MRM screen for neurotoxicity in preclinical models with the capability for extrapolation to clinical analyses using classical MRI. PMID- 10668996 TI - Stress proteins as molecular markers of neurotoxicity. AB - In response to many environmental and pathophysiologic stressful stimuli, cells undergo a stress response characterized by induction of a variety of proteins, including the heat shock protein family. The inducible heat shock protein 70 (hsp70) is believed to participate in an array of cellular activities, including cytoprotection. Normal brain cells have little detectable hsp70 RNA or protein. However, following a stressful condition hsp70 mRNA and protein are induced in different cell types depending on the severity and the nature of the stimulus. The induction of hsp70 protein correlates with the regional and cellular vulnerability to a particular injury as identified by standard histologic methods. The pattern of hsp70 expression differs in response to various neurotoxic stimuli, including hyperthermia, ischemia, seizures, hemorrhage, and N methyl-D-aspartate receptor antagonist administration. Hsp70 expression is a useful marker of cellular injury and may help to identify previously unrecognized areas of vulnerability in the nervous system after a neurotoxic stimulus. Hsp70 may also play a neuroprotective role in the brain. PMID- 10668997 TI - Teased-fiber technique for peripheral myelinated nerves: methodology and interpretation. AB - Teased-fiber technique is the best approach for studying peripheral myelinated nerve fibers in their continuity. It enables the assessment of size of myelin segments formed by Schwann cells and characterization of pathologic changes affecting the internodia, the paranodal regions, and the invested axons. Fiber teasing is performed on prestained proximodistally oriented portions of peripheral nerves. Specimens about 10 mm long are stained for 24-48 hours in Sudan black and then transferred to glycerin, where, using a pair of fine forceps and a stereomicroscope, they are separated into smaller fiber bundles from which single fibers are isolated. The work is performed on a glass slide with an adhesive surface (albuminized or "superfrost"), on which the fibers are placed in strict proximodistal orientation. Following drying in an oven, the slides are mounted with glycerin-gelatine (same as used for frozen sections). The changes, when present, can usually be recognized during the preparation, but fibers are reexamined and changes confirmed in mounted slides. Photographic reconstruction of the fibers facilitates their assessment and enables the documentation of findings. The teased-fiber technique is auxiliary to histopathology, and to limit the workload and save costs, it can be performed on only a few specimens selected for better characterization of changes recognized or suspected in tissue sections. In particular, segmental demyelination and early stages of Wallerian or secondary axonal degeneration can be recognized in teased fibers. Segmental demyelination is characterized by loss of fully myelinated segments and their replacement by newly formed short and thin segments, remyelinating the preserved axon. The early stage of secondary axonal degeneration is recognized by formation of ovoidal fiber fragments in the midinternodal region. PMID- 10668998 TI - Practical aspects of neuropathology: a technical guide for working with the nervous system. AB - Toxicologic pathologists are evaluating tissues from the central and peripheral nervous systems with increasing frequency. This change is being driven by recently established regulatory guidelines and intense interest in developing pharmaceutical compounds to treat various nervous system disorders. However, morphologic evaluation of the nervous system by light or electron microscopy requires special understanding and effort. Here, we review the general concepts of fixation for the nervous system, explain perfusion procedures for optimal preservation, and provide information on handling tissues to avoid artifacts. In general, fixation with aldehydes is recommended for nervous tissue (a combination of paraformaldehyde and glutaraldehyde is preferred). Electron microscopic studies require fixatives of the highest purity possible, typically paraformaldehyde prepared fresh from powder mixed with high-grade glutaraldehyde. The final osmolality of the solution should be slightly hypertonic, in the range of 400-600 mOsmol. Slight hypertonicity is very important and will facilitate maintenance of vascular distention during whole-body perfusion, which is the best method for producing high-quality tissue preparations. Special effort is necessary for handling nervous tissue in a way that minimizes artifacts because chemical fixation is not completed immediately following the perfusion. These technical details should help toxicologic pathologists in their efforts to work with the nervous system, thereby increasing their effectiveness in supporting safety characterization of new test materials undergoing toxicologic assessments. PMID- 10668999 TI - ECOs, FOBs, and UFOs: making sense of observational data. AB - Systematic observations of rat behavior are required for both standard subchronic safety studies and for neurotoxicity studies. The requirements specify subjective out-of-cage observations (eg, posture, gait, and reactivity to various stimuli such as, auditory, tactile, and noxious) using defined scales. Measurement of forelimb/hind limb grip strength, landing foot splay, and locomotor activity are also required. The observational endpoints are organized into a battery, eg, the Environmental Protection Agency functional observational battery (FOB) or expanded clinical observations (ECO). Functional and neuropathologic data are most easily integrated when the functional endpoints are organized as a neurologic exam (ie, each endpoint has a known anatomical basis and there are sufficient endpoints to cover the nervous system). Current batteries do not constitute a neurologic exam. Although ECOs and FOBs contain some components of a neurologic exam (ie, observations of gait, response to pinch), the anatomic basis for other components (eg, hind limb splay) is poorly defined. And although some functions (eg, somatomotor) are well characterized by current batteries, others (eg, vision, somatosensation) are evaluated less effectively. The measurement of locomotor activity in a novel environment is one of the most problematic parts of current functional testing batteries, although contemporary technology may provide opportunities for improving this test. The influence of inherent limitations of functional test methods is magnified by factors associated with testing for neurotoxicant-related effects during safety studies. First, most personnel at contract laboratories have little or no formal training in conducting and interpreting a neurologic examination. Second, most neurotoxicant related lesions are bilateral, which paradoxically may produce more subtle effects than unilateral lesions. Third, most chemicals will be tested only once, and sponsors are reluctant to evaluate results in "real time" and amend protocols to add endpoints (eg, neurophysiological tests) to clarify functional effects. Pathologists should have realistic expectations about the opportunities for integrating functional and neuropathologic findings. PMID- 10669000 TI - Ototoxicity: an argument for evaluation of the cochlea in safety testing in animals. AB - The cochlea is one of the more common targets for toxic effects, yet current toxicologic screening in animals does not routinely evaluate the cochlea as a potential target organ. Although histopathologic sections are routinely taken from the eye and the optic nerve and tract and most studies include at least 1 section through the nasal cavity and olfactory mucosa, the cochlea is not histopathologically examined in routine toxicity studies. Unfortunately, routine clinical examinations frequently miss ototoxicity because rodents and other species can lose most of their high-frequency hearing and still respond to most ambient noises. Ototoxicity as a deficiency in toxicologic screening can be remedied by using well-established histopathologic and behavioral methods or electrophysiologic methods, such as brain stem auditory evoked responses (BAERs). Once the equipment is in place, BAERs can be obtained quickly and easily for ototoxicity screening (approximately 15 minutes for paired testing of 2 rats and 30 minutes each for dogs). BAERs also can be used in virtually all mammalian species. Three or 4 probe frequencies (eg, 4, 8, 16, and 32 kHz), representing different areas of the cochlea, can be tested in a few minutes with subcutaneous electrodes under short-acting chemorestraint. Given the availability of several approaches to screening for ototoxicity and the importance of the auditory function in human health, safety tests of chemicals and drugs should include an effective screening test for ototoxicity. PMID- 10669001 TI - Characterization of carbon disulfide neurotoxicity in C57BL6 mice: behavioral, morphologic, and molecular effects. AB - Female C57BL6 mice were exposed to 0 or 800 ppm carbon disulfide (CS2), 6 h/d, 5 d/wk for 20 weeks. The neurologic function of all mice was assessed once at the end of exposures using a functional observational battery. General health effects included a decrease in body weight gain, piloerection, hunched body posture, and ptosis. Treatment-related effects included altered gait (uncoordinated placement of hind limbs and ataxia) and impaired function on an inverted screen test. In addition, rearing and locomotor movement were decreased in treated mice. Focal to multifocal axonal swelling was seen predominantly in the muscular branch of the posterior tibial nerve, and occasionally giant axonal swelling was detected in the lumbar segment of the spinal cord. Electron microscopic examination revealed swollen axons with massive accumulation of neurofilament proteins within the axoplasm. Covalent cross-linking of erythrocyte spectrin (surrogate protein to neurofilament protein) was demonstrated in mice exposed to CS2 but not in mice receiving filtered air. These data provide supportive evidence that covalent cross-linking of neurofilament proteins is a significant feature of the axonal swellings in mice produced by inhalation exposure to CS2. PMID- 10669002 TI - The role of developmental neurotoxicology studies in risk assessment. AB - A number of questions have been raised about the use of the US Environmental Protection Agency's Developmental Neurotoxicity Testing Guideline (DNTG) in the hazard identification of chemicals. The applicability and sensitivity of animal tests in the DNTG relative to human developmental neurotoxicity have recently been questioned. In a workshop held in 1989, participants compared the effects of several known developmental neurotoxicants in humans and animal models and concluded that the DNTG would have detected known human developmental neurotoxicants. They also concluded that although procedural differences may differ in the testing of humans and animals, the neurobiologic functions (ie, autonomic, sensory, motor, and cognitive) affected by chemical exposure were similar. In cases where the DNTG has been compared with other measures of reproductive and developmental toxicity, the DNTG has been relatively sensitive and specific. To date, DNTGs have been required 12 times, for 9 pesticides and 3 solvents. The sensitivity of the measures in the DNTG relative to other measures of developmental and adult toxicity supports the continued use of the DNTG in risk assessment. PMID- 10669003 TI - Morphometric analysis of the developing rat brain. AB - In 2 studies, a method of linear morphometry was applied to regulatory developmental neurotoxicity studies in the rat. The first study involved the development of the brain during postnatal days (PNDs) 7-63, and the second involved the effects of 8 mg/kg i.p. trimethyltin chloride (TMT) to rats at PND 8, with morphometry performed at PNDs 12 and 24. The results of the TMT linear morphometry were compared with those from stereologic counting of neurons in the cerebral cortex, piriform cortex, and hippocampus. Stereology produces more meaningful data than simple linear morphometry for use in the regulatory assessment of the developmental neurotoxicity potential of compounds. PMID- 10669004 TI - Phenotype vs genotype in the evolution of astrocytic brain tumors. AB - Astrocytic brain tumors are the most frequent human gliomas and they include a wide range of neoplasms with distinct clinical, histopathologic, and genetic features. Diffuse astrocytomas are predominantly located in the cerebral hemispheres of adults and have an inherent tendency to progress to anaplastic astrocytoma and (secondary) glioblastoma. The majority of glioblastomas develop de novo (primary glioblastomas), without an identifiable less-malignant precursor lesion. These subtypes of glioblastoma evolve through different genetic pathways, affect patients at different ages, and are likely to differ in their responses to therapy. Primary glioblastomas occur in older patients and typically show epidermal growth factor receptor (EGFR) overexpression, PTEN mutations, p16 deletions, and, less frequently, MDM2 amplification. Secondary glioblastomas develop in younger patients and often contain TP53 mutations as their earliest detectable alteration. Morphologic variants of glioblastoma were shown to have intermediate clinical and genetic profiles. The giant cell glioblastoma clinically and genetically occupies a hybrid position between primary (de novo) and secondary glioblastomas. Gliosarcomas show identical gene mutations in the gliomatous and sarcomatous tumor components, which strongly supports the concept that there is a monoclonal origin for gliosarcomas and an evolution of the sarcomatous component due to aberrant mesenchymal differentiation in a highly malignant astrocytic neoplasm. PMID- 10669005 TI - A mouse model for glioma: biology, pathology, and therapeutic opportunities. AB - The epidermal growth factor receptor (EGFR) gene is amplified or mutated in 30 50% of human glioblastoma multiforme. These mutations are usually associated with deletions of the INK4a-ARF locus, which encodes 2 gene products (p16INK4a and p19ARF) involved in cell cycle arrest and apoptosis. We have investigated the role of EGFR mutation in gliomagenesis using avian retroviral vectors to transfer a mutant EGFR gene to glial precursors and astrocytes in transgenic mice. These mice express tv-a, a gene encoding the retrovirus receptor TVA, which is under the control of brain cell type-specific promoters. We demonstrate that expression of a constitutively active, mutant form of EGFR in cells in the glial lineage can induce lesions with many similarities to human gliomas, including increased cell density, vascular proliferation, and immunohistochemical staining for glial fibrillary acidic protein (GFAP) and nestin. We also demonstrate that primary astrocytes cultured from transgenic mice expressing tv-a from the GFAP promoter are efficiently infected in culture, and such genetically modified cell cultures can be tumorigenic in nude mice. The combinations of genetic lesions (eg, mutated EGFR, INK4a-/-) leading to tumor formation in these 2 mouse systems are similar to those found in human gliomas. These genetically defined animal models for gliomas will allow for the testing of therapies that are targeted specifically at the gene products involved in the pathogenesis of gliomas. PMID- 10669006 TI - Morphologic characterization of spontaneous nervous system tumors in mice and rats. AB - Spontaneous rodent nervous system tumors, in comparison to those of man, are less well differentiated. Among the central nervous system (CNS) tumors, the "embryonic" forms (medulloblastoma, pineoblastoma) occur both in rodents and humans, whereas the human "adult" forms (gliomas, ependymomas, meningiomas) have fewer counterparts in rodents. In general, the incidence of spontaneous CNS tumors is higher in rats (>1%) than in mice (>0.001%). A characteristic rat CNS tumor is the granular cell tumor. Usually it is associated with the meninges, and most meningeal tumors in rats seem to be totally or at least partly composed of granular cells, which have eosinophilic granular cytoplasm, are periodic acid Schiff reaction (PAS)-positive, and contain lysosomes. Such tumors are frequently found on the cerebellar surface or at the brain basis. Rat astrocytomas are diffuse, frequently multifocal, and they invade perivascular spaces and meninges. The neoplastic cells with round to oval nuclei and indistinct cytoplasm grow around preexisting neurons, producing satellitosis. In large tumors, there are necrotic areas surrounded by palisading cells. Extensive damage of brain tissue is associated with the presence of scavenger cells that react positively with histiocytic/macrophage markers. The neoplastic astrocytes do not stain positively for glial fibrillary acidic protein; they probably represent an immature phenotype. In contrast to neoplastic oligodendroglia, they bind the lectin RCA-1. Astrocytomas are frequently located in the brain stem, especially the basal ganglia. Rat oligodendroglial tumors are well circumscribed and frequently grow in the walls of brain ventricles. Their cells have water-clear cytoplasm and round, dark-staining nuclei. Atypical vascular endothelial proliferation occurs, especially at the tumor periphery. Occasionally in the oligodendrogliomas, primitive glial elements with large nuclei occur in the form of cell groups that form rows and circles. Primitive neuroectodermal tumors of rats, such as pineal tumors or medulloblastomas, appear to have features similar to those found in man. In mice, the meningeal tumors are mostly devoid of granular cells and the astrocytomas are similar to those occurring in rats, whereas spontaneous oligodendrogliomas are observed extremely rarely. Tumorlike lesions, such as lipomatous hamartomas or epidermoid cysts, are occasionally encountered in the mouse CNS. It is suggested that we classify rodent CNS lesions as "low grade" and "high grade" rather than as "benign" and "malignant." The size of CNS tumors is generally related to their malignancy. Tumors of the peripheral nervous system are schwannomas and neurofibromas or neurofibrosarcomas consisting of Schwann cells, fibroblasts, and perineural cells. Well-differentiated schwannomas are characterized by S-100 positivity and the presence of basement membrane. They show either Antoni A pattern with fusiform palisading cells or Antoni B pattern, which is sparsely cellular and has a clear matrix. The rat develops specific forms of schwannomas in the areas of the submandibular salivary gland, the external ear, the orbit, and the endocardium. Spontaneous ganglioneuromas occur in the rat adrenal medulla or thyroid gland. Compared to experimentally induced neoplasms, the spontaneous tumors of the rodent nervous system are poor and impractical models of human disease, although they may serve as general indicators of the carcinogenic potential of tested chemicals. PMID- 10669007 TI - Evaluation of ENU-induced gliomas in rats: nomenclature, immunochemistry, and malignancy. AB - Rats developed mixed gliomas, oligodendrogliomas, and a few astrocytomas in response to transplacental ethylnitrosourea. The neoplastic cell composition of mixed gliomas must be defined; this study required a 20-80% admixture of neoplastic astrocytes and oligodendroglia for the diagnosis of mixed glioma. A battery of immunoantibodies, including Leu-7, S-100, and vimentin, were helpful in classifying rat gliomas, and the histologic features of each tumor type are described. Other brain tumor characteristics that may decide the outcome of carcinogenicity studies include incidence, multiplicity, latency, fatality, size, and malignancy. The size of tumors was determined by measuring their 3 dimensional volumes. Brain tumor volume was found to be highly correlated with malignancy and fatality. Systematic evaluation of the malignancy of brain tumors is an important but often overlooked adjunct method of measuring the effectiveness of a carcinogen. A system to estimate malignancy, one that grades 9 tumor characteristics and weights, each according to clinical outcome, was developed. It was found that mixed gliomas grew larger, had a shorter latency, and were significantly more malignant than were other gliomas. PMID- 10669008 TI - Tumors of the nervous system in carcinogenic hazard identification. AB - In the absence of adequate data on humans, it is biologically plausible and prudent to regard agents and mixtures for which there is sufficient evidence of carcinogenicity in experimental animals, usually rats and mice, as if they presented a carcinogenic risk to humans. Prediction of cancer sites in humans from bioassay data in rodents is much less certain, however, regardless of organ or tissue. For tumors of the nervous system, there is practically no basis for judging the validity of such predictions, as only ionizing radiation is known to cause tumors of the central nervous system (CNS) in humans. Brain tumors are relatively uncommon findings in bioassays and are rare in untreated rodents, even in rats, which appear to be the most susceptible species. However, CNS tumors have been readily induced in rodents by systemic exposures to some chemicals, notably N-nitrosoalkylureas and other alkylating agents and certain alkyl hydrazine derivatives. CNS tumors in rodents have played a significant role in carcinogenic hazard evaluations of several other chemicals, including acrylonitrile, ethylene oxide, and acrylamide, and have been implicated as part of the tumor spectrum induced by vinyl chloride and certain inorganic lead compounds. In some of these evaluations, it is not certain that all tumors diagnosed as primary brain tumors were correctly identified. Diagnostic difficulties have been presented by undifferentiated small-cell tumors that may invade the brain, including carcinomas of the nasal cavity and undifferentiated schwannomas arising in cranial nerve ganglia, and by the difficulty of reliably distinguishing between focal reactive gliosis and early glial neoplasms. The most striking experimental finding regarding the induction by chemicals of tumors of the nervous system is the dramatically greater susceptibility of the fetal and neonatal nervous system to some carcinogens, as compared with the susceptibility of the nervous system in adults of the same species. PMID- 10669009 TI - Plant cyanogenic glycosides. AB - The cyanogenic glycosides belong to the products of secondary metabolism, to the natural products of plants. These compounds are composed of an alpha hydroxynitrile type aglycone and of a sugar moiety (mostly D-glucose). The distribution of the cyanogenic glycosides (CGs) in the plant kingdom is relatively wide, the number of CG-containing taxa is at least 2500, and a lot of such taxa belong to families Fabaceae, Rosaceae, Linaceae, Compositae and others. Different methods of determination are discussed (including the indirect classical photometrical and the new direct chromatographic ones). The genetic control of cyanogenesis has no unique mechanism, the plants show variation in the amount of the produced HCN. The production of HCN depends on both the biosynthesis of CGs and on the existence (or absence) of its degrading enzymes. The biosynthetic precursors of the CGs are different L-amino acids, these are hydroxylated then the N-hydroxylamino acids are converted to aldoximes, these are turned into nitriles. The last ones are hydroxylated to alpha-hydroxynitriles and then they are glycosilated to CGs. The generation of HCN from CGs is a two step process involving a deglycosilation and a cleavage of the molecule (regulated by beta-glucosidase and alpha-hydroxynitrilase). The tissue level compartmentalisation of CGs and their hydrolysing enzymes prevents large-scale hydrolysis in intact plant tissue. The actual level of CGs is determined by various factors both developmental and ecological ones, which are reviewed too. The last part of the present work demonstrates the biological roles of CGs in plant physiological processes and in plant defence mechanisms as well. The effect of CGs (HCN) on different animals, the symptoms of poisonings are discussed to cows, sheep, donkeys, horses and chicks. Finally, the poisonous effects of cassava (Manihot esculenta) roots are summarised on experimental animals and on the human organism. PMID- 10669010 TI - First report on the distribution of orally administered microcystin-LR in mouse tissue using an immunostaining method. AB - The purpose of this study was to investigate the distribution of microcystin-LR (MCLR) orally administered to mice using an immunostaining method. MCLR was orally dosed at 500 microg/kg to aged Balb/C and ICR mice and their lethality was 23.9%. The former was more sensitive to MCLR than the latter, suggesting that oral toxicity by MCLR is related to the animal strains tested, although the pathological and immunostaining changes were essentially the same in both strains. According to this method the distribution of MCLR and related compounds were indicated as the red staining. Particularly, livers of dead aged mice were intensively stained. The main route of absorption was considered to be the small intestine because the villi contained a large amount of MCLR in both surface epithelial cells and lamina propria, resulting in erosion. The absorbed MCLR was contained in blood plasma and moved to the liver, lung, and heart, and finally to capillaries of the whole body. Excretion of MCLR was shown in the mucous from goblet cells in both the small intestine and large intestine. PMID- 10669011 TI - Some toxic and enzymatic activities of Bothrops ammodytoides (yarara nata) venom. AB - Bothrops ammodytoides, the smallest representative of this genus, is found only in Argentina. Venom was extracted from thirty adult specimens (35-70 cm in length, 90-300 g in weight) captured in the Province of Buenos Aires and kept in captivity. Venom yield was 3-30 mg. SDS-PAGE showed strong bands at 14.0; 23-25; 45; 54 and 63 kDa and weak bands at 17.0; 30.0; 40.0 and 85.0 kDa. Toxic activities were: LD50 (intravenous, mice) 0.5+/-0.2 microg/g; minimal procoagulant dose on human plasma (MPD-P) 35+/-2 mg/l; and minimal defibrinogenating dose (MDD, mice) 6-12 microg. Hemorrhagic and/or necrotic activities appear to play a major role in lethality; minimal hemorrhagic dose (MHD, mice) is 10+/-2 microg/g and minimal necrotizing dose (MND, mice) is 38+/-5 microg. The LD50, MPD-P and MND are among the lowest in venoms from Bothrops species found in Argentina. B. ammodytoides venom exhibited high proteolytic and phospholipase A2 activities. Most of the B. ammodytoides venom components cross react with Bivalent Bothropic antivenom (Instituto Nacional de Produccion de Biologicos ANLIS Dr. G. Malbrin, against B. alternatus and B. neuwiedii venoms). One ml of antivenom neutralizes 1.2 mg of B. ammodytoides venom. PMID- 10669012 TI - Purification and characterization of BaH4, a hemorrhagic metalloproteinase from the venom of the snake Bothrops asper. AB - A hemorrhagic metalloproteinase, named BaH4, was isolated from the venom of the snake Bothrops asper by a combination of ion-exchange chromatography on DEAE Sepharose and gel filtration on Sephacryl S-200. BaH4 is a 69 kDa protein with a pI of 5.3. It was recognized by antibodies raised against hemorrhagic metalloproteinase BaH1 isolated from B. asper venom, with a reaction of partial immunologic identity. BaH4 shows proteolytic activity on biotinylated casein, hide powder azure and fibrin, although having lower activity than crude B. asper venom and metalloproteinase BaP1 isolated from the same venom. BaH4 hydrolyzed fibronectin, laminin and type IV collagen in vitro, albeit at a relatively high enzyme:substrate ratio. Proteolytic activity was inhibited by chelating agents and 2-mercaptoethanol, but not by soybean trypsin inhibitor. Prominent hemorrhage developed in gastrocnemius and cremaster muscles after administration of BaH4. Moreover, it induced lethality in mice after intravenous injection, with an LD50 of 0.37 microg/g. Histological observations showed conspicuous pulmonary hemorrhage when the enzyme was injected intravenously. BaH4 is a hemorrhagic metalloproteinase which may play a relevant role in local and systemic bleeding characteristic of B. asper envenomations. PMID- 10669013 TI - An investigation of the biological activity of bullrout (Notesthes robusta) venom. AB - Bullrout envenomation is known to cause intense pain. Crude bullrout venom and venom fractions were assessed for protease, hyaluronidase, phospholipase and hemolytic activities, reactivity with stonefish antivenom, lethality to brine shrimp and ability to elicit pain in human subjects. Compared with venom obtained from frozen specimens, live fish venom-milking techniques rendered greater venom potency and improved storage characteristics. Although mild proteolytic and hemolytic activity was observed, crude venom demonstrated no hyaluronidase or phospholipase A2 activity, did not affect brine shrimp, or show antigenicity with stonefish antivenom. A single venom protein isolated from bullrout venom is attributed with causing pain in human subjects. The sensations elicited by this novel algesic protein are consistent with chemical stimulation of polymodal nociceptors. PMID- 10669015 TI - KTX3, the kaliotoxin from Buthus occitanus tunetanus scorpion venom: one of an extensive family of peptidyl ligands of potassium channels. AB - A new ligand of the K+ channels sensitive to KTX was purified from the venom of Buthus occitanus tunetanus, using two steps of high-performance-liquid chromatography and by following its ability to compete with [125I]-KTX for binding to the KTX receptor on rat brain synaptosomes. Amino-acid analysis, amino acid sequencing and mass spectroscopy defined this new ligand. KTX3, as a 37 amino acid peptide, with three disulfide bridges. Its sequence shares 76% identity with KTX. The main differences between the two peptides are in the N terminal region and the residue position 34 located in the region involved in channel recognition. These differences may explain the 5-fold lower binding affinity of KTX3, IC50=50 pM, than KTX to rat brain synaptosomes. Specific antibodies raised against KTX (1-37) were not able to recognize KTX3. PMID- 10669014 TI - Isolation and structures of grammistins, peptide toxins from the skin secretion of the soapfish Grammistes sexlineatus. AB - Two peptide toxins (named grammistins Gs 1 and Gs 2) with hemolytic and ichthyotoxic activities were isolated from the skin secretion of the soapfish Grammistes sexlineatus. Grammistin Gs 2 showed 6-11 x higher hemolytic activity and 10x higher ichthyotoxicity than grammistin Gs 1. The complete amino acid sequences of Gs 1 comprising 25 residues and Gs 2 comprising 24 residues were determined. Although a search by the database failed to find any homologous toxins from other sources, the grammistins were similar in secondary structures as well as biological activities to the two classes of peptide toxins, melittin from the bee venom and pardaxins from the skin secretion of two species of soles. CD experiments and helical wheel projections showed that the grammistins were randomly coiled in distilled water but formed amphiphilic alpha-helices in the presence of SDS micelles. In addition, they were found to be surface seeking peptides by the Eisenberg plot and assumed to exist as aggregates of 3-4 molecules. Interestingly, grammistin Gs 2 is much more abundant in amphiphilic alpha-helices and much higher in biological activities than melittin and pardaxins as well as grammistin Cs 1. PMID- 10669016 TI - Induction of neutralizing antibodies against Tityus serrulatus toxins by immunization with a recombinant nontoxic protein. AB - An immunogenic nontoxic protein (TsNTxP) was purified from the venom of the scorpion Tityus serrulatus (Ts). This peptide is composed of 63 amino acid residues with a high degree of structural homology with the toxins isolated from Ts. The nucleotide sequence of the gene that encodes TsNTxP was obtained and also showed a high degree of similarity with genes encoding Tityus toxins [Guatimosim, S.C.F., Prado, V.F., Diniz, C.R., Chavez-Olortegui, C.. Kalapothakis, E., 1999. Molecular cloning and genomic analysis of TsNTxP; an immunogenic protein from Tityus serrulatus scorpion venom. Toxicon 37, 507-517]. In the present study the TsNTxP gene was expressed in E. coli BL21DE3 cells as a fusion protein with maltose-binding protein. The recombinant protein (TsNTxPrec) was purified by affinity chromatography and used as an immunogen in rabbits. The antigenic specificity of anti-TsNTxPrec antibodies was compared by an enzyme-linked immunosorbent assay using TsNTxP, TstFG50 (the fraction of Ts venom that represents most of the toxicity of the crude venom) and the crude venom, to coat microtitration plates. Anti-TsNTxPrec antibodies had a comparable high cross reactivity for all antigens tested. Concentrations of Ts venom equivalent to 20 LD50 were effectively neutralized by 1 ml of the anti-TsNTxPrec serum. This result provides basic data for the use of such recombinant scorpion protein as an immunogen in the development of antivenoms for clinical use. PMID- 10669017 TI - Bleeding from the small intestine caused by aplysiatoxin, the causative agent of the red alga Gracilaria coronopifolia poisoning. AB - The cause of death by aplysiatoxin poisoning was bleeding from the small intestine in mice. The pathological changes related to the cause and progression of bleeding were studied morphologically. Bleeding from the capillaries was observed 60 min after i.p. treatment at 250 microg/kg, and this was preceded by dilatation of the lymphatic vessel and congestion of capillaries in the lamina propria from 10 min after the injection. At 100 microg/kg i.v., the target vessels were in the lung, where fibrin deposition was observed in the dilated pulmonary artery, and blood flowed out through a gap in the artery. Then, in the small intestine, similar changes appeared to have occurred, and bleeding was induced in two characteristic ways, one through deposition of fibrin in the lumen and the other via distension of the capillary wall. PMID- 10669018 TI - Paralizing activity of the Parawixia bistriata crude venom in termites: a new bioassay. AB - Spider venoms have high specificity to neuronal elements. Therefore, the use of venom has been important in the characterisation of mammal and insect nervous systems. The evaluation of insect paralysis has been an important tool for distinguishing the biological effects of venom. In this study we describe the paralysing effect of a spider crude venom (Parawixia bistriata) in termites, utilising a new bioassay. The crude venom of P. bistriata caused an irreversible and dose-dependent paralysis in the animals in the following doses: 2.10(-5) U; 2.10(-4) U; 2.10(-3) U; 2.10(-2) U and 0.12 U (1 U = 1 gland). This bioassay will allow for easy and direct evaluation of biological effects from different venoms and purified fractions. PMID- 10669019 TI - Neutralization of the hemorrhagic activity of Bothrops and Lachesis snake venoms by a monoclonal antibody against mutalysin-II. AB - One mAb reactive with mutalysin-II, a hemorrhagic metalloproteinase isolated from Lachesis muta muta venom, was produced in mice immunized with L. m. muta venom. Indirect ELISA was employed to compare the antigenic cross-reactivity among the venoms from Bothrops snakes. The mAb anti-mutalysin-II efficiently neutralized the hemorrhagic effect of both mutalysin-II and L. m. muta crude venom. Furthermore, the mAb were cross-reactive with B. alternatus, B. atrox, B. itapetiningae, B. jararaca and B. neuwiedii and showed variable potencies in neutralizing the hemorrhagic activity of several bothropic venoms. PMID- 10669020 TI - Bibliography of toxinology. PMID- 10669021 TI - Are we as toxinologists overlooking vouchers? PMID- 10669022 TI - The effect of Portuguese Man-of-war (Physalia physalis) venom on calcium, sodium and potassium fluxes of cultured embryonic chick heart cells. AB - Portuguese Man-of-war venom markedly increases calcium (45Ca2+) influx into primary, cultured, embryonic chick heart cells. This action is dose-dependent, but is unaffected by organic calcium blockers (diltiazem, verapamil, nifedipine, nimodipine and mibefradil). On the other hand, certain trivalent (La3+, Gd3+) and divalent (Zn2+, Ni2+, Cu2+, Mn2+) metals inhibit venom-induced calcium influx. Sodium (22Na+) influx into chick heart cells is also significantly increased by Man-of-war venom. Flecainide does not block venom-induced sodium influx. The efflux of the potassium analogue, 86Rb+, from heart cells is also significantly increased by the venom. The venom, however, has little or no effect on rubidium (86Rb+) or 2-deoxy-D-[2-3H] glucose influx. PMID- 10669023 TI - Occurrence of the toxin dehydroabietic acid in Salmonella typhimurium. AB - Many strains of Salmonella typhimurium studied in our lab demonstrated marked differences in the pathogenicity for guinea pig, chicken and Hela cells. As a result, a pathogenic strain of S. typhimurium, strain 9SR2, was evaluated for lipophilic components that may be associated with virulence using gas chromatography/mass spectrometry. The hydroxylated fatty acids 2 hydroxytetradecanoic acid (2-OH-14:0) and 3-hydroxytetradecanoic acid (3-OH-14:0) often present in lipid A, a potent endotoxin, were observed as their methyl esters. The cyclic fatty acids methylene-hexadecanoic acid (C17delta) and methyleneoctadecanoic acid (C19delta) also were detected. The nephrotoxic and neurotoxic diterpenoid resin acid, dehydroabietic acid, was observed for the first time from S. typhimurium in both the total lipid and diglyceride fractions and determined as its methyl ester at m/z 314.2246. Due to its previously established toxicity, dehydroabietic acid may be a factor associated with virulence of S. typhimurium. PMID- 10669024 TI - An ultrasensitive competitive binding assay for the detection of toxins affecting protein phosphatases. AB - An ultrasensitive assay is described for microcystin-LR and other substances (microcystins, nodularin, okadaic acid, calyculin A, tautomycin) which block the active site of protein phosphatases (PP) 1 and 2A. The assay is based on competition between the unknown sample and [125I]microcystin-YR for binding to the catalytic subunit of PP2A. The PP2A-bound [125I]microcystin-YR was stable (half-time of dissociation = 1.8 h), allowing non-bound [125I]microcystin-YR to be removed by Sephadex G-50 size-exclusion chromatography. Compared to current assays based on inhibition of protein phosphatase activity the present assay was more robust against interference (from fluoride, ATP, histone, and casein), and had an even better sensitivity. The detection limit was below 50 pM (2.5 fmol) for nodularin and microcystin-LR, and below 200 pM (10 fmol) for okadaic acid. The method was used successfully to detect extremely low concentrations of either microcystin or nodularin in drinking water or seawater, and okadaic acid in shellfish extract. PMID- 10669025 TI - Purification and cDNA cloning of an insecticidal protein from the venom of the scorpion Orthochirus scrobiculosus. AB - Injection of crude venom from the scorpion Orthochirus scrobiculosus into larvae of Heliothis virescens (Lepidoptera: Noctuidae) caused trembling and uncoordinated movement before development of a progressive and prolonged flaccid paralysis. The isolation of the toxin (OsI-1) responsible for this effect of O. scrobiclosus venom is described. The molecular mass of OsI-1 toxin was 6994 Da, as determined by desorption mass spectroscopy. The complete primary structure of OsI-1 was deduced from the sequence of cDNA clones obtained by rapid amplification of cDNA ends (RACE) PCR. Comparison of the deduced amino acid sequence of OsI-1 with those of other insecticidal scorpion toxins indicates that it is a sodium (Na+) channel active depressant insect-selective toxin. The analysis of amino acid sequence of the toxin in conjunction with mass spectroscopy data indicates post-translational modification in maturation with the removal of 3 C-terminal amino acids and amidation of the C-terminus. PMID- 10669026 TI - Characterisation of antibacterial activity of peptides isolated from the venom of the spider Cupiennius salei (Araneae: Ctenidae). AB - The characterisation of the antimicrobial activity of five antibacterial peptides, isolated from the venom of the neotropical wandering spider Cupiennius salei is reported here. The peptides have a molecular mass, determined by electrospray ionisation-mass spectrometry, between 3-4 kDa. Minimal inhibitory concentrations against five different bacteria species were determined by a liquid growth inhibition assay. All five peptides showed minimal inhibitory concentrations that are comparable to those of other known antibacterial peptides, like insect defensins and cecropins, found in the last years in a large diversity of animals. The peptides are supposed to lyse the cells by formation of either distinct channels or pores, but their mode of action is not yet revealed. PMID- 10669027 TI - Development of sensitive colorimetric capture elisas for Clostridium botulinum neurotoxin serotypes A and B. AB - Sensitive and specific enzyme-linked immunosorbent assays were developed to detect Clostridium botulinum neurotoxin serotypes A (BoNT A) and B (BoNT B) in assay buffer and human serum. The assay is based upon affinity-purified horse polyclonal antibodies directed against the approximately 50 kDa C-fragments of each toxin. Standard curves were linear over the range of 0.1-10 ng mL. Detection was possible at 0.2 ng mL (20 pg/well) and accurate quantitation at 0.5 ng/mL (50 pg well) in assay buffer and 10% human serum. Variations between triplicates was typically 5-10%. Less than 1% cross reactivity occurred between other serotypes when each assay was performed against serotypes A, B and E. When tested against toxins complexed to their associated nontoxic proteins, interference was absent (BoNT B) or < 25% (BoNT A). These assays demonstrate sensitivity close to that of the mouse bioassay without the use of animals and in a much simpler format than other reported assays of similar sensitivity. PMID- 10669028 TI - Dose-dependent cardiovascular and neuromuscular effects of stonefish (Synanceja trachynis) venom. AB - There has been recent debate regarding the labile nature of stonefish venoms and the pharmacology of their breakdown products. The present study examined the cardiovascular and neuromuscular effects of lyophilised venom, and conducted a preliminary investigation of freshly milked venom. Lyophilised venom (20 microg/ml) caused endothelium-dependent relaxation in rat aortae that was abolished by atropine (0.1 microM). In contrast, an endothelium-independent contractile response occurred in porcine coronary arteries. However, in the presence of atropine (10 nM), this became a relaxation response which was attenuated by the B2 antagonist FR-173657 (0.1 microM) or by a combination of idazoxan (1 microM) and propranolol (1 microM). In rat isolated atria, lyophilised venom (4 microg/ml) caused a biphasic inotropic response consisting of an initial decrease, and then increase, in force which were attenuated by atropine (0.5 microM) and propranolol (5 microM), respectively. The increase in force produced by venom was unaffected by reserpine pre-treatment suggesting a direct action at adrenoceptors. In the anaesthetised rat, lyophilised venom (1 300 microg/kg, i.v.), caused a dose-dependent depressor response, with a subsequent pressor response at higher concentrations (30-300 microg/kg, i.v.). In the presence of atropine (1 mg/kg, i.v.), the depressor response to venom was abolished, a transient pressor response unmasked and the secondary pressor response augmented. In the additional presence of prazosin (50 microg/kg, i.v.), the transient pressor response was abolished and the secondary pressor response attenuated. Lyophilised venom had no significant effect on nerve-evoked (10 microg/ml) or directly-evoked (100 microg/ml) twitches of the chick biventer cervicis muscle preparation. Milked venom (1 microl/ml) caused a biphasic response (i.e., an initial relaxation followed by contraction) in rat aortae, a contraction in porcine coronary arteries, complete cessation of rat isolated atrial activity and markedly inhibited both nerve-evoked and directly-evoked twitches of the chick biventer cervicis muscle preparation. In the anaesthetised rat, milked venom (15 microl/kg, i.v.) caused immediate cardiovascular collapse. It appears that the cardiovascular effects of stonefish venom are mediated by a dose-dependent action at muscarinic receptors and adrenoceptors. PMID- 10669029 TI - Clinical features and management of Hadronyche envenomation in man. AB - Using case reports and a review of the literature, the clinical features of envenomation by the genus of Australian funnel web spiders known as Hadronyche, are characterised. Five cases are reported here, including the first life threatening envenomation by Hadronyche species 14 (the Port Macquarie funnel web). Two severe envenomations by Hadronyche cerberea (the Southern Tree funnel web) and one each by Hadronyche formidabilis (the Northern Tree funnel web) and Hadronyche infensa (the Darling Downs funnel web) are also described. The clinical experience of the authors' provided the five cases described in detail one of which has previously been reported in brief. Eight cases of Hadronyche envenomation from the literature (Medline 1966-1998 and Embase 1980-1998) were analysed in order to draw comparisons between this syndrome and the well described envenomation syndrome of Atrax robustus (the Sydney funnel web). Reports of funnel web spider antivenom use to Commonwealth Serum Laboratories (CSL) between 1995 and June 1998 were also examined. The biology of these dangerous spiders, their geographic distribution, venom characteristics and management issues are addressed. It is concluded that bites from at least six Hadronyche species have produced a life-threatening envenomation syndrome clinically indistinguishable from that of Atrax robustus. Atrax robustus derived antivenom is effective although antivenom requirements may be greater than for Atrax envenomation. Antivenom supplies are limited and sufficient stocks to treat a severe envenomation are unlikely to be found in any one institution. Pressure immobilisation first aid is effective in delaying onset of envenomation, may enhance local inactivation of venom and early removal can result in rapid clinical deterioration. PMID- 10669030 TI - Isolation of a funnel-web spider polypeptide with homology to mamba intestinal toxin 1 and the embryonic head inducer Dickkopf-1. AB - We have isolated and determined the amino acid sequence of a novel peptide component from the venom of the Australian funnel-web spider Hadronyche versuta. This 68-residue toxin, ACTX-Hvf17, does not function like classical neurotoxins in modulating ion channel function as evidenced by its lack of insecticidal activity and its inability to affect vertebrate smooth or skeletal muscle contractility. The peptide shows significant sequence homology with mamba intestinal toxin 1 (MIT1) and to a lesser extent with a variety of colipases. The strong structural homology between MIT1 and porcine colipase leads us to propose that ACTX-Hvf17 also adopts the MIT1/colipase three-dimensional fold. However, we show that ACTX-Hvf17 has no colipase activity and does not stimulate muscle contractility like MITI. We also show that MIT1 and ACTX-Hvf17 display significant sequence homology with the C-terminal cysteine-rich domain of the Dickkopf-1 family of proteins that induce head formation in developing embryos, which leads us to propose that this domain of Dickkopf-1 also adopts the MIT1 colipase fold. PMID- 10669031 TI - Quantification of crotamine, a small basic myotoxin, in South American rattlesnake (Crotalus durissus terrificus) venom by enzyme-linked immunosorbent assay with parallel-lines analysis. AB - Intraspecific variation in Crotalus durissus terrificus venom composition was studied in relation to crotamine activity. Crotamine induces paralysis in extension of hind legs of mice and myonecrosis in skeletal muscle cells. To determine whether the venom of crotamine-negative rattlesnake contains a quantity of myotoxin incapable of inducing paralysis, we have developed a very sensitivity immunological assay method, an enzyme-linked immunoabsorbent assay (ELISA), capable of detecting 0.6 ng of purified crotamine. The parallel-lines analysis of ELISA data showed to be useful because it shows the reliability of the experimental conditions. A variation in the amount of myotoxin in the crotamine positive venom was observed, but not less than 0.1 mg of crotamine per mg of venom. It was not possible to detect it in crotamine-negative venom even at high venom concentrations. PMID- 10669032 TI - Regional and accelerated molecular evolution in group I snake venom gland phospholipase A2 isozymes. AB - In accordance with detection of a few phospholipase A2 (PLA2) isozyme genes by Southern blot analysis, only two cDNAs, named NnkPLA-I , and NnkPLA-II, encoding group I PLA2s, NnkPLA-I and NnkPLA-II, respectively, were isolated from the venom gland cDNA library of Elapinae Naja naja kaouthia of Malaysia. NnkPLA-I and NnkPLA-II showed four amino acid substitutions, all of which were brought about by single nucleotide substitution. No existence of clones encoding CM-II and CM III, PLA2 isozymes which had been isolated from the venom of N. naja kaouthia of Thailand, in Malaysian N. naja kaouthia venom gland cDNA library was verified by dot blot hybridization analysis with particular probes. NnkPLA-I and NnkPLA-II differed from CM-II and CM-III with four and two amino acid substitutions, respectively, suggesting that their molecular evolution is regional. The comparison of NnkPLA-I, NnkPLA-II and cDNAs encoding other group I snake venom gland PLA2s indicated that the 5'- and 3'-untranslated regions are more conserved than the mature protein-coding region and that the number of nucleotide substitutions per nonsynonymous site is almost equal to that per synonymous site in the protein-coding region, suggesting that accelerated evolution has occurred in group I venom gland PLA2s possibly to acquire new physiological functions. PMID- 10669033 TI - Purification and some properties of a tetrodotoxin binding protein from the blood plasma of kusafugu, Takifugu niphobles. AB - A tetrodotoxin binding protein has been purified from the plasma of the puffer fish kusafugu, Takifugu niphobles, through DEAE-cellulose treatment, ammonium sulfate fractionation, Sephadex gelfiltrations and Sephacryl S-200 and Cellulofine A-500 column chromatography. Final purification by HPLC on a TSK G 3000 SL column yielded a protein which showed only a single protein peak. The molecular weight of the protein was estimated to be 116,000 and 91,000 by SDS PAGE and mass spectrometry, respectively. A blast search on the amino-terminal amino acid sequence of the purified protein revealed that the protein had no homology to any other protein on data base. PMID- 10669034 TI - Bibliography of toxinology. PMID- 10669035 TI - Evaluation of NOx in the cardiovascular system: relationship to NO-related compounds in vivo. AB - Diverse attention should be paid to evaluating NOx (NO2- and NO3-) in plasma as an index of endothelial nitric oxide (NO) formation in vivo. Nitric oxide, which subsequently appears as NOx, originates from different types of NO synthase and from nonenzymatic reactions. NOx also comes from exogenous sources such as food and gastrointestinal microorganisms. The fate of the NO incorporated into activation of guanylate cyclase, formation of nitrosyl hemoglobin (or nitrosohemoglobin), nitrosothiols, peroxynitrite and its derivatives and other possible compounds is not clear at present. However, some of these compounds would produce NOx as by-products or as final products through metabolism. Therefore, plasma NOx contains information about these pathways, although how extensively these factors contribute to plasma NOx has not been quantitatively defined. A theoretical simulation of NOx in the systemic circulation indicates that only small changes are expected by inhibition or stimulation of endothelial NO production. Measuring NOx production during coronary circulation has the advantage that some degree of NOx accumulation is expected from intact endothelial cells because an excretion system is absent in the heart. PMID- 10669036 TI - Receptor-mediated modulation of voltage-dependent Ca2+ channels via heterotrimeric G-proteins in neurons. AB - The activity of voltage-dependent Ca2+ channels is highly regulated by neurotransmitter receptors coupled to heterotrimeric G-proteins. In the expression studies using cloned Ca2+ channel subunits, it has been clarified that the main mechanism of the inhibition of N-type channel current is mediated directly by G-protein betagamma subunits in a membrane-delimited and voltage sensitive manner. In addition, recent studies have also clarified that N-type channels are modulated by several G-protein alpha subunits in different ways. Among them, G(alpha o) mediates a voltage-resistant inhibition of N-type current by neurotransmitters. This type of inhibition is more apparent in the case of P/Q type channels in both native cells and expression systems. Moreover, other G protein subunits, such as G(alpha q) and G(alpha s), also seem to regulate N-type channels in a membrane-delimited manner. The fine tunings of Ca2+ channel activity by intracellular proteins have physiological and pathological meanings in the regulation of Ca2+ influx into excitable cells by neurotransmitters and pharmacological implications as novel drug targets for controlling Ca2+ influx. PMID- 10669037 TI - Evaluation of the histamine H1-antagonist-induced place preference in rats. AB - The place preferences by some histamine H1 antagonists, such as tripelennamine, optical isomers of chlorpheniramine (dl-, d- and l-forms) and pyrilamine, in rats were evaluated with the conditioned place preference paradigm. In the present study, tripelennamine and all of the optical isomers of chlorpheniramine, but not pyrilamine, produced a significant place preference. The degree of the place preference induced by optical isomers of chlorpheniramine (6.0 mg/kg) did not correlate with the H1-antagonistic potency of these drugs, suggesting that H1 antagonist-induced place preferences are not mediated by H1-receptor blockade. The tripelennamine (3.0 mg/kg)- and dl-chlorpheniramine (6.0 mg/kg)-induced place preferences were completely abolished by pretreatment with the dopamine D1 receptor antagonist SCH23390 (0.05 mg/kg). Furthermore, the doses of H1 antagonists that induced a place preference significantly reduced the levels of DOPAC, which may be mediated by inhibition of dopamine uptake, in the limbic forebrain (including the nucleus accumbens and olfactory tubercle). These results suggest that some H1 antagonists induce rewarding effects, which may be mediated by the activation of dopamine D1 receptors, followed by the inhibition of dopamine uptake. PMID- 10669038 TI - Different effects of trypsin inhibitors on intestinal gene expression of secretin and on pancreatic bicarbonate secretion in CCK-A-receptor-deficient rats. AB - The effects of oral administration of two synthetic trypsin inhibitors (camostate and ONO-3403) and soybean trypsin inhibitor (SBTI) on cholecystokinin (CCK), secretin gene expression and pancreatic secretion were examined in CCK-A-receptor deficient (OLETF) rats. The rats were fed chow containing 0.1% trypsin inhibitors for 7 days. To examine pancreatic secretion, the rats were prepared with cannulae to drain the bile and pancreatic juice separately, a duodenal cannula and an external jugular vein cannula. The animals were maintained in Bollman cages and the experiments were conducted 4 days after surgery. The levels of CCK mRNA were significantly increased by each treatment. The levels of secretin mRNA were significantly increased by camostate and SBTI, but not by ONO-3403. Bicarbonate secretion was significantly increased in rats treated with camostate and ONO 3403, but not SBTI, while protein secretion was not affected by any treatment. These observations suggest that increased bicarbonate secretion produced by synthetic trypsin inhibitors in CCK-A-receptor-deficient rats may not be due to secretin but due to ONO-3403 in the circulation. PMID- 10669039 TI - The lipophilic properties of angiotensin I-converting enzyme inhibitors do not influence their diffusion through cultured endothelium. AB - The background for these investigations was the discovery that formation of angiotensin II by the renin angiotensin system can take place in extravascular tissues (e.g., cardiomyocytes and neurons) and within single cells. Consequently, the question arose about whether such tissue-based systems might be differentially influenced by angiotensin I-converting enzyme (ACE) inhibitors with distinct physicochemical properties. Therefore, the aim of this study was to investigate how the membrane penetration of various ACE inhibitors depends on their lipophilia. All diacid forms of ACE inhibitors are dissociated at a pH of 7.4 and scarcely extractable into octanol (extraction coefficient < 10%). In contrast, the extraction coefficients of the parent substances showed marked differences in the following order of increasing lipophilia: enalapril = perindopril < captopril = ceranapril < ramipril < quinapril < HOE288 = zofenopril < fosinopril < HOE065. For selected substances, the kinetics of diffusion through a monolayer of cultured bovine aortic endothelium were determined. The diffusion rates (expressed as half lives) of captopril (59.6 min), enalapril (53.4 min), enalaprilat (50.8 min), ramipril (56.9 min) and ramiprilat (51.1 min) are similar indicating: 1) that penetration is independent on lipophilia and 2) that endothelium constitutes no specific barrier for the passage of ACE inhibitors into the vessel wall. PMID- 10669040 TI - Sensitization of the adenylyl cyclase system in cloned kappa-opioid receptor transfected cells following sustained agonist treatment: A chimeric study using G protein alpha(i)2/alpha(q) subunits. AB - Chronic and/or sustained opioid treatment has been shown to result in development of sensitization of the adenylyl cyclase (AC) system or cAMP overshoot. In this study, we investigated the molecular mechanism responsible for sensitization of the AC system using CHO cells co-expressing cloned kappa-opioid receptor and some chimeric G protein alpha(i2)/alpha(q) subunits. In CHO cells co-expressing the kappa-opioid receptor and pertussis toxin-insensitive chimeric alpha(i2)/alpha(q) subunits with alpha(i2) residues Met244-Asn331, despite pretreatment with pertussis toxin, acute treatment with the kappa-opioid-receptor-selective agonist U69,593 suppressed forskolin-stimulated cAMP accumulation, while sustained treatment with U69,593 (4 h) induced cAMP overshoot over the naive level by the kappa-opioid-receptor-selective antagonist norbinaltorphimine (sensitization of the AC system). On the other hand, in CHO cells co-expressing the kappa-opioid receptor and pertussis toxin-insensitive chimeric alpha(i2)/alpha(q) subunits without alpha(i2) residues Met244-Asn331, pretreatment with pertussis toxin completely blocked these acute and sustained effects of U69,593 on cAMP accumulation. These results suggested that the presence of the specific region of alpha(i2) (Met244-Asn331), which was reported to be responsible for the inhibition of AC, and continuous inhibition of AC by alpha(i2) is necessary for the development of sensitization. PMID- 10669041 TI - The role of endothelium-derived nitric oxide in relaxations to levcromakalim in the rat aorta. AB - The present study was designed to examine the role of basally released nitric oxide in relaxations to an ATP-sensitive K+ channel opener. Whether relaxations to levcromakalim are modulated by endothelial removal or the inhibitors of vasodilator effects of endothelium-derived nitric oxide, were investigated in the rat aorta. During contractions to phenylephrine (3 x 10(-7) to 10(-6) M), levcromakalim (10(-8) to 10(-5) M) or a nitric oxide donor, 1-hydroxy-2-oxo-3-(N methyl-3-aminopropyl)-3-methyl-1-triazene (NOC-7, 10(-9) to 10(-5) M), was added in a cumulative fashion. Relaxations to levcromakalim (10(-8) to 10(-5) M) were significantly reduced by the endothelium-removal. In aortas with endothelium, relaxations in response to levcromakalim were decreased by selective inhibitors of nitric oxide synthase (N(G)-nitro-L-arginine methyl ester, 10(-4) M) and soluble guanylate cyclase (1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one; ODQ, 10( 5) M) and a scavenger of nitric oxide (carboxy-PTIO, 10(-3) M). Relaxations to levcromakalim in aortas treated with these inhibitors are comparable to those seen in aortas without endothelium. KCl (30 mM) and an ATP-sensitive K+ channel inhibitor, glibenclamide (10(-5) M), abolished relaxations to levcromakalim in aortas with or without endothelium, whereas glibenclamide did not alter relaxations to NOC-7 (10(-9) to 10(-5) M) in aortas without endothelium. These results suggest that in rat aortas, inhibition of vasodilator effects of basally released nitric oxide can reduce relaxations via ATP-sensitive K+ channels, although these channels do not mediate relaxations to exogenously applied nitric oxide. PMID- 10669042 TI - Involvement of brain serotonergic terminals in the antinociceptive action of peripherally applied calcitonin. AB - We investigated the antinociceptive effect of systemic injection of calcitonin and its mechanisms of action in rats. Subcutaneous injection of [Asu(1,7)]eel calcitonin (ECT, 4 U x kg(-1) x day(-1)) daily for 7 days suppressed nociceptive hypersensitivity induced by formalin (and by carrageenan); the effect was gradually increased by the repeated injections and significant effects were observed after administration for more than 4 days. The antinociceptive action of ECT (4 U x kg(-1) x day(-1) for 7 days) was inhibited by intracerebroventricular injection of the serotonergic neurotoxin 5,7-dihydroxytryptamine and serotonin receptor antagonists methiothepin, cyproheptadine and ketanserin; methysergide showed an inhibitory tendency. Intrathecal injections of 5,7-dihydroxytryptamine, methiothepin, cyproheptadine and ketanserin were without effects on the ECT action. The results suggest the involvement of serotonin in the brain, but not in the spinal cord, in the ECT antinociception. Intracerebroventricular or intrathecal injection of the catecholaminergic neurotoxin 6-hydroxydopamine and intracerebroventricular injection of the alpha-adrenoceptor antagonist phentolamine were also without effects on the ECT action. A subcutaneous infusion of the opioid receptor antagonist naloxone inhibited the antinociceptive action of morphine, but not that of ECT. Thus, adrenergic and opioidergic systems may not play important roles in the ECT antinociception. The present results suggest that repeated systemic injection of ECT produces analgesia and that the brain serotonergic terminals are involved in this action. PMID- 10669043 TI - Endogenous ATP released by electrical field stimulation causes contraction via P2x- and P2y-purinoceptors in the isolated tail artery of rats. AB - Electrical field stimulation (EFS) caused contraction of isolated tail arteries of rats. The EFS-induced contraction showed frequency-dependence and was entirely abolished by the sodium channel blocker tetrodotoxin (1 x 10(-7) M). The EFS induced (at 20 Hz) contraction was reduced by about 60% in the presence of phentolamine (1 x 10(-6) M). Therefore, later experiments were carried out in the presence of phentolamine. Pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) (1 x 10(-8)-1 x 10(-6) M) and basilen blue E-3G (3 x 10(-5)-5 x 10(-5) M), P2-receptor antagonists, significantly inhibited the contraction evoked by EFS. In addition, PPADS significantly inhibited the contractions induced by ATP (1 x 10(-4) M) and a selective P2x-receptor agonist, alpha,beta-methylene ATP (1 x 10(-6) M). In contrast, basilen blue E-3G did not inhibit alpha,beta-methylene ATP-induced contraction. The ecto-ATPase activator apyrase (5 and 10 U/ml) significantly reduced the EFS-induced contractions. These findings suggest that endogenous ATP released by EFS causes contractions of rat tail artery via both the P2x-receptors and P2y-receptors. PMID- 10669044 TI - Effect of zaldaride maleate, an antidiarrheal compound, on visceral pain reflex induced by small intestinal distention in anesthetized rats. AB - Using distention of the small intestine as a visceral pain model, we investigated the effect of zaldaride maleate (ZAL), a selective inhibitor of calmodulin, on the depressor response. In pentobarbital-anesthetized rats, small intestine distention was induced by rapid application of intraluminal pressures of 40 cmH2O causing a reflex fall in arterial blood pressure. The depressor response to intestinal distention was abolished by intraperitoneal administration of capsaicin (5 mg/rat), which depletes neuropeptides such as substance P from the sensory neurons, on the mesenteric stalk and by neonatal pretreatment with capsaicin (50 mg/kg, s.c.). Morphine (20 mg/kg, s.c.) reduced the depressor response following intestinal distention. At doses of 3 mg/kg (i.v.) and higher, ZAL significantly reduced depressor response. The effect of morphine was reversed by naloxone (5 mg/kg, i.v.); the effect of ZAL was not affected. These results suggest that ZAL helps reduce the visceral pain induced by noxious stimulus and that the antinociceptive effect of ZAL is not mediated by opioid receptors. PMID- 10669045 TI - Structure and sequence of the mouse V1a and V1b vasopressin receptor genes. AB - The structural organization and 5'-flanking region of the mouse V1a and V1b vasopressin receptor genes were determined. The mouse V1a receptor gene was located within an 8-kb XbaI fragment, and the mouse V1b receptor gene was located within a 14-kb EcoRV fragment. Both genes were comprised of two coding exons that were separated by a 2.3-kb and a 8.0-kb intron, respectively, located before the respective seventh transmembrane domain of the receptor sequence. The availability of these genes would allow us to study the functional role of V1a and V1b receptors by disrupting the gene in mice. PMID- 10669046 TI - Influence of chronic treatment with imipramine on mRNA levels in rat brain: elevation of glyceraldehyde-3-phosphate dehydrogenase levels. AB - The differential display method was used to identify the intrinsic factor that changes its mRNA expression level in rat brain after a 14-day oral administration of 20 mg/kg imipramine. The expression of a 180-bp band was markedly enhanced by imipramine. The results of sequencing and a data base search revealed that the isolated clone was glyceraldehyde-3-phosphate dehydrogenase (GAPDH) with a one base difference. Enhancement of the expression by imipramine was observed in the amygdala. Quantitative PCR showed that imipramine treatment significantly elevated the GAPDH/beta-actin ratio in the cortex. These findings suggest that long-term treatment with imipramine stimulates GAPDH mRNA expression. PMID- 10669048 TI - Effect of benidipine on depolarizing stimulation-induced increase of intracellular calcium concentration in cultured mouse hippocampal neurons. AB - We examined the effect of benidipine, a 1,4-dihydropyridine calcium channel blocker, on depolarizing stimulation-induced increases of intracellular calcium concentration ([Ca2+]i) in cultured mouse hippocampal neurons in comparison with those of nicardipine and nilvadipine. Benidipine (0.1-10 microM) inhibited the [Ca2+]i increase compared with the no drug control response. This effect was stronger than those of nicardipine and nilvadipine. The inhibitory effect of benidipine lasted even after washing out the drug for 125 min, while those of nicardipine and nilvadipine disappeared more rapidly. This is the first report that demonstrates that benidipine inhibits the [Ca2+]i increase in the neuron itself. PMID- 10669047 TI - Antidiarrheal effects of zaldaride maleate after oral, intravenous and subcutaneous administration to rats. AB - The antidiarrheal action of zaldaride maleate (ZAL) after oral, intravenous and subcutaneous administration was examined to determine whether ZAL acts systemically or locally in the intestine of rats. Oral administration of ZAL inhibited castor oil- and 16,16-dimethyl prostaglandin E2-induced diarrhea; however, intravenous or subcutaneous administration of ZAL was ineffective. When ZAL was orally administered, the area under the plasma concentration time curve of the compound was lower than that of ZAL following intravenous or subcutaneous administration at the maximum doses studied. The antidiarrheal effect of ZAL was not dependent on its plasma concentration level. These results suggest that ZAL acts locally in the intestinal tract in rats. PMID- 10669049 TI - Inhibition of human chymase by suramin. AB - Chymase is a chymotrypsin-like protease localized in mast cells in complexes with heparin. In the present study, we demonstrated that suramin, a hexasulfonated naphthylurea used as an anti-cancer drug, inhibits the activity of purified human chymase in vitro. The inhibition was ionic-strength-dependent. It was observed that suramin competed with heparin-Sepharose gel for binding to chymase and the inhibition of chymase activity by suramin was partially impaired by heparin. Our results show that suramin may become a prototype of a new type of chymase inhibitor because of its unique character. PMID- 10669050 TI - An approach to the medical marijuana controversy. AB - The use of smoked marijuana as a therapeutic agent is presently a matter of considerable debate in the United States. Many people suffering from a variety of disorders maintain that it is necessary for their adequate treatment. Yet, the evidence to support claims is insufficient for FDA approval. An interim solution is proposed which would allow patients referred by their physicians to participate in a 6-month program of legal marijuana availability, similar to the 'compassionate IND' program of a number of years ago. A technique similar to that used for post-marketing surveillance is proposed for obtaining quantitative data for a limited number of potential indications. These are: (1) nausea and vomiting associated with cancer chemotherapy or other causes, (2) weight loss associated with debilitating illnesses, (3) spasticity secondary to neurological diseases, and (4) chronic pain syndromes. PMID- 10669051 TI - A comprehensive guide to the application of contingency management procedures in clinical settings. AB - Controlled clinical research has demonstrated the efficacy of contingency management procedures in treating substance use disorders. Now is the time to begin introducing these procedures into standard clinical practice. This article reviews the rationale of contingency management interventions and provides a review of representative scientific work in the area. It also discusses behaviors that can be modified, reinforcers that can be used, and behavioral principles that can be adapted to improve outcomes. This paper provides practical advice and a guideline for clinicians and researchers to use when designing and administering contingency management interventions. The recommendations are based on empirically validated manipulations. Areas in which more research is needed are suggested as well. PMID- 10669052 TI - Gender differences in hostility of opioid-dependent outpatients: role in early treatment termination. AB - The purpose of this study was to evaluate gender differences in hostility and the role of hostility in predicting early treatment termination of opioid-dependent outpatients. Demographic characteristics and Addiction Severity Index (ASI) ratings were collected from 104 patients (68 males and 36 females) at intake to a buprenorphine treatment program. Hostility was assessed using the Buss-Durkee Hostility Scale. Compared to male opioid-dependent patients, females scored significantly higher on this scale. Early treatment termination was defined as remaining in treatment < 30 days, and 13% percent of males and 25% of females were classified as early terminators. Stepwise logistic regression identified predictors of early treatment termination. Severity of legal and employment problems and the interaction between hostility and gender predicted early treatment termination status. Patients with less severe legal problems and patients with greater employment problems were more likely to terminate early from treatment. Higher levels of hostility predicted early treatment termination of female patients, but hostility levels were not associated with treatment termination in male patients. Results from this study show that female heroin addicts have high levels of hostility and suggest that hostility may be an important predictor of premature discharge from opioid substitution programs, especially among women. PMID- 10669053 TI - Illicit cocaine use patterns in intravenous-naive cocaine users following investigational intravenous cocaine administration. AB - This study evaluated whether cocaine use patterns changed following investigational intravenous cocaine administration to intravenous-naive cocaine users. Subjects were respondents to a follow-up survey who had participated in one to three intravenous double-blind cocaine (0.2 or 0.4 mg/kg) administration studies. The group included healthy men (n = 17) and women (n = 8) with histories of occasional cocaine use (lifetime self-reported use of 12+/-12 (mean +/- S.D.) exposures, primarily via nasal insufflation) who were recontacted an average of 39 weeks (range 7-107 weeks) after study participation. The recontacted group constituted 45% of the total eligible sample of 55 subjects. Baseline demographics for the recontacted and non-recontacted (n = 30) samples were similar, suggesting that the recontacted sample was representative of the group as a whole. Investigational cocaine exposure did not induce adverse health events in any subject. Self-reported cocaine use estimates obtained at follow-up were compared to baseline estimates obtained with identical questionnaires and were highly concordant (Spearman rank correlation p = 0.52 and 0.78, respectively; P < 0.02 and < 0.0002, respectively). This suggests that participants provided stable and reliable reports of cocaine use. No subject reported either illicit intravenous cocaine use or altered frequency of illicit cocaine use by the customary route after investigational intravenous cocaine exposure. These data suggest that illicit cocaine use frequencies and routes of administration are not altered following investigational intravenous cocaine administration to healthy, occasional cocaine users. PMID- 10669054 TI - Predictors of expressed partner and non-partner violence among patients in substance abuse treatment. AB - This study examined reports of expressed partner and non-partner violence among men (n = 126) and women (n = 126) in the 12 months prior to substance abuse treatment. Rates of violence were 57% for partner, 53% for non-partner, and 75% collapsing across partner and non-partner relationships. Factors associated with partner and non-partner violence severity differed substantially. Partner violence was predicted by age, marital status, and drug problem severity. Non partner violence was predicted by gender, income, alcohol and drug problem severity. The results highlight that individuals in substance abuse treatment are at high risk for violence, and targeted screening and intervention approaches should be routine in addictions treatment. PMID- 10669055 TI - Contingency management in outpatient methadone treatment: a meta-analysis. AB - A meta-analysis was conducted on contingency management interventions in outpatient methadone treatment settings. The outcome measure of interest was drug use during treatment, as detected through urinalysis. The results confirm that contingency management is effective in reducing supplemental drug use for these patients. The analysis of behavioral interventions yielded an overall effect size (r) of 0.25 based on 30 studies. Significant moderators of outcomes included type of reinforcement provided, time to reinforcement delivery, the drug targeted for behavioral change, number of urine specimens collected per week, and type of subject assignment. These factors represent important considerations for reducing drug use during treatment. PMID- 10669056 TI - Stress induced spontaneous recurrence of methamphetamine psychosis: the relation between stressful experiences and sensitivity to stress. AB - We examined increased sensitivity to stress in relation to spontaneous recurrences of methamphetamine (MAP) psychosis (i.e., flashbacks). Plasma monoamine metabolite levels were assayed in: 26 flashbackers, of whom 11 were on neuroleptics before and during the study, and the other 15 received neuroleptics in the course of the study; 18 non-flashbackers with a history of MAP psychosis; eight subjects with persistent MAP psychosis; and 23 MAP user and 11 non-user controls. The 26 flashbackers had experienced stressful events and/or MAP-induced fear-related psychotic symptoms during previous MAP use. Mild psychosocial stressors then triggered flashbacks. During flashbacks plasma norepinephrine levels increased markedly; among the flashbackers, those with a history of stressful events, whether or not they had experienced fear-related symptoms, showed a further increase in 3-methoxytyramine levels. Stressful experiences, together with MAP use, may therefore induce sensitization to stress associated with noradrenergic hyperactivity, involving increased dopamine release, and so triggering flashbacks. PMID- 10669057 TI - The role of stimulated lipid peroxidation and impaired calcium sequestration in the enhancement of cocaine induced hepatotoxicity by ethanol. AB - The purpose of this study was to investigate possible mechanism of cocaine induced hepatotoxicity and its potentiation by ethanol in mice. Ethanol (2 g/kg) and/or cocaine (25 mg/kg) injections were given as binge model (five injections in 3 days). Cocaine administration with or without ethanol caused an increase in lipid peroxidation in liver homogenate and its subcellular fractions. The greatest increases were observed in mitochondrial fraction following cocaine plus ethanol treatment. Also, glutathione (GSH) levels were increased in liver homogenate and its mitochondrial fractions after cocaine and cocaine plus ethanol treatment. Microsomal calcium sequestration was found to decrease in all treatments. These results suggest that increased lipid peroxidation and decreased microsomal calcium sequestration in the liver may play a possible role cocaine induced hepatotoxicity and its potentiation by ethanol. PMID- 10669058 TI - Inpatient opiate detoxification in Geneva: follow-up at 1 and 6 months. AB - The aim of this study was to identify predictors of treatment success and of relapse, 1 and 6 months after inpatient opiate detoxification in an 8-bed unit in Geneva. Of all 73 patients admitted between June 1994 and June 1995, a majority (73%) successfully finished opiate detoxification. Detoxification was performed mainly with methadone tapering; the average duration of hospitalisation was 15 days. Factors associated with treatment failure were: cocaine abuse, presence of legal problems, and short duration of hospital stay. After 1 month, 65% of the patients were using drugs (half of them were dependent again, half of them had used occasionally) and 35% were completely abstinent (21% when excluding those in residential treatment). Predictors of rapid relapse were cocaine abuse and little concern with own psychological situation at baseline. After 6 months, 50% were physically dependent again, 13% had lapsed occasionally, 37% were abstinent (28% when excluding those in residential treatment). Only high benzodiazepine use at baseline was associated with medium term abstinence. Addiction severity index composite scores had considerably improved between baseline and 6 months. Prevention of relapse to opiate use after inpatient detoxification, especially for those with a concurrent cocaine abuse, should be improved. PMID- 10669059 TI - Dissociation of physical abstinence signs from changes in extracellular dopamine in the nucleus accumbens and in the prefrontal cortex of nicotine dependent rats. AB - The aim of the present study was to investigate the relationship between physical abstinence and changes in dopamine release in the nucleus accumbens and in the medial prefrontal cortex induced by mecamylamine and naloxone in rats chronically exposed to nicotine. The rats were implanted with osmotic minipumps (Alzet) delivering nicotine tartrate at a rate of 9 mg/kg/day (3.16 mg of free base) and 8 days later with a dialysis probe in the nucleus accumbens or in the medial prefrontal cortex. Steady-state dopamine output from the nucleus accumbens of the rats implanted with nicotine minipumps was higher than that of sham implanted rats; no differences were observed in the prefrontal cortex. In nicotine but not in sham implanted rats mecamylamine (1 mg/kg s.c.) precipitated a physical abstinence syndrome and brought dopamine output back to control values in the nucleus accumbens. In contrast mecamylamine (1 mg/kg s.c.) increased dopamine output in the medial prefrontal cortex of nicotine but not sham-implanted rats. Naloxone (2 mg/kg) precipitated a physical abstinence syndrome qualitatively similar to that produced by mecamylamine but failed to modify extracellular dopamine in the nucleus accumbens or in the prefrontal cortex of nicotine implanted and sham-implanted rats. The results indicate that the mesolimbic and mesocortical dopamine system undergo opposite changes during mecamylamine precipitated abstinence in rats chronically exposed to nicotine and that physical abstinence signs can be dissociated from changes in dopamine transmission. PMID- 10669060 TI - A within-subject comparison of three different schedules of reinforcement of drug abstinence using cigarette smoking as an exemplar. AB - In this experiment we compared three different schedules of reinforcement for promoting and sustaining short-term drug abstinence. For pragmatic reasons, cigarette smoking was studied as an exemplar of drug self-administration. The three schedules studied were a fixed magnitude of reinforcement for abstinence, a progressive increase in magnitude of reinforcement for abstinence with a reset contingency for drug use, and a progressive increase in magnitude of reinforcement for abstinence without a reset contingency. Eighteen cigarette smokers experienced the three schedules in a counterbalanced order. Each schedule was in effect for 5 consecutive days (M-F), during which time abstinence was reinforced according to the different schedules of reinforcement. The total amount of reinforcement (money) available was the same during each condition. The progressive magnitude with a reset schedule was more effective than the other two schedules in sustaining an initial period of abstinence. These results systematically replicate and extend those from prior studies demonstrating the efficacy of schedules incorporating a progressively increasing magnitude of reinforcement with a reset contingency for sustaining initial drug abstinence, and demonstrate the importance of the reset contingency to that effect. PMID- 10669061 TI - Drug use, self report and urinalysis. AB - Stimulated by the ever present demand to consider the financial implications in management decisions, this study examines the use of urinalysis and self-report in the treatment of drug users, to question if urinalysis, rather than being a routine investigation, could be used with greater discrimination without jeopardising its effectiveness. It concludes that urinalysis remains of importance, as an adjunct to self-report, in providing information and in the treatment of drug users. Suggestions are made as to how it might be used more selectively in treatment based on a clinical knowledge of individual patients and their progress in treatment. However further research is needed to support and define this more clearly. PMID- 10669062 TI - Methadone, methadone treatment and non-fatal overdose. AB - This paper provides a mainly qualitative investigation of the role of methadone and methadone treatment in non-fatal illicit drug overdose. During 1997 and 1998, semi-structured interviews were conducted with 33 individuals in six hospital accident and emergency departments in two Scottish cities. The research identified four overdose situations related to methadone/methadone treatment. These were: (1) topping up a legitimate methadone prescription; (2) abusing another's methadone prescription; (3) preferring illegal drugs to prescribed methadone; and (4) failing to obtain prescribed methadone. The implications of these findings for methadone treatment policy and practice are discussed. PMID- 10669063 TI - Effects of prolonged ethanol exposure on neurophysiological measures during an associative learning paradigm. AB - Long-term ethanol exposure has been reported to produce electrophysiological and cognitive impairments in some alcoholics. This study assessed the effects of chronic ethanol exposure on neurophysiological indices of associative learning in rats. Male Wistar rats (46) were exposed to ethanol vapor (EtOH group) or air (control group) for 6 consecutive weeks. After the animals were withdrawn from ethanol, electrodes were implanted in the frontal and parietal cortices and in the amygdala. Following a prolonged abstinence from ethanol (10-15 weeks), rats were exposed to a classical conditioning paradigm in which a food pellet was paired with the presentation of an auditory stimulus. During the first five sessions (conditioning phase), food pellet presentation was paired with the presentation of an infrequently presented tone. During the second five sessions (extinction phase), the association between food pellet presentation and the infrequently presented tone was weakened by no longer presenting food pellets following the infrequent tone. During selected test sessions, event-related brain potentials (ERPs) elicited by each tone (i.e. food-paired tone, non-paired tone) were recorded and analyzed. These analyses revealed differences in ERP responses between the groups. The latency of the N1 and P2 ERP components in the cortex of the control group, but not the EtOH group, increased during sessions when the association between food pellet delivery and tone presentation was being established or extinguished. These data support the hypothesis that chronic ethanol treatment results in a loss of responsivity in ERP components sensitive to changes in food-tone associations, even following a prolonged period of withdrawal from ethanol. PMID- 10669064 TI - Estimates of total alcohol consumption in Russia, 1980-1994. AB - It is important to estimate real alcohol consumption in Russia in the past 15 years because large quantities, and in recent years the bulk, of consumed alcohol evades state registration. In this study, trends in total consumption are estimated indirectly using an indicator of alcohol-related harm derived from forensic reports on accidental and violent deaths (Nemtsov, A.V., 1998. Anti alcohol campaign, consequences, and actual alcohol consumption in Moscow, Addiction, in press). Using blood alcohol coefficients (BAC), the regression coefficients are estimated using the ratio of BAC-positive and BAC-negative accidental and violent deaths in 1983-1986 in connection with the Moscow anti alcohol campaign; the coefficients are then used to estimate consumption as a total sum of legal sales of alcoholic beverages and illegal spirits made from sugar. Data were obtained of violent BAC-positive and BAC-negative deaths in 17 25 oblasts of Russia in 1981-1994. The derived estimates indicate that in 1984 consumption exceeded 14 l per capita, per annum (10.5 registered + 4.2 unregistered); that consumption fell to 10.8 1 in 1986 (during the anti-alcohol campaign), and that it then began to rise owing to a sharp increase in estimated unregistered alcohol consumption-climbing to 13.6 l (5.0 + 8.6) in 1993, and followed by a slight decline to 13.3 in 1994. These estimates are broadly consistent with estimates that have been made using unrelated methods, thus allowing some confidence in the overall picture obtained. Russia probably remains one of the heaviest-drinking countries in Europe. The reasons why indicators of alcohol-related harm in the mid-1990s exceed those recorded before the anti alcohol campaign remain to be clarified. PMID- 10669065 TI - Efficacy of daily and alternate-day dosing regimens with the combination buprenorphine-naloxone tablet. AB - This study evaluated the efficacy of a combination tablet formulation of buprenorphine containing 8 mg of buprenorphine and 2 mg of naloxone for every other day treatment and whether increasing the daily maintenance dose was essential for maintaining an efficacious alternate-day treatment. Twenty-six opioid-dependent outpatients completing a 16-day baseline entered a double-blind, placebo-controlled, triple crossover trial. Twenty-one days of daily combination tablet administration were compared to two different 21-day periods of alternate day buprenorphine administration where subjects received either 8 or 16 mg of the combination tablet every other day with placebo on the interposed day. Fifty-four percent (14/26) of subjects completed the study, but only two subjects dropped out during the 16-mg alternate-day condition. Rates of medication compliance, illicit opioid use and subject- and observer-rated measures of opioid effects did not distinguish daily from alternate-day treatments in subjects completing the study. However, pupillary diameter was significantly increased during 8-mg alternate-day compared to the 8-mg daily or 16-mg alternate-day treatment. These data replicate earlier findings describing the acceptability of alternate-day buprenorphine treatment using multiples of the daily maintenance dose and extend these findings by establishing the clinical efficacy of daily and alternate-day dosing regimens with the combination buprenorphine naloxone tablet. This study also suggests slightly improved outcomes during alternate-day treatment using multiples of the daily dose. PMID- 10669066 TI - HIV-1 RNA load in needles/syringes from shooting galleries in Miami: a preliminary laboratory report. AB - We quantified HIV-1 RNA load in rinses from needles/syringes (N/S) obtained at shooting galleries in Miami and also analyzed the rinses for antibodies for viral proteins. In rinses from 36 N/S that contained visible blood, 14 (39%) had detectable amounts of HIV-1 RNA. Numbers of copies of HIV-1 RNA ranged from the detection limit (400 copies/ml) to 268,000 copies/ml. We also detected antibodies to HIV-1 polypeptides in 34/36 (94%) of rinses from visibly contaminated N/S using Western blots specific for the HIV-1 proteins. No antibodies were detected in laboratory rinses from six visibly clean needles. The presence of HIV-1 RNA in N/S is an important indication of the risk created by N/S sharing as well as by shared paraphernalia and wash waters by injecting drug users. PMID- 10669067 TI - Reversibility of morphine effects on phagocytosis by murine macrophages. AB - Proneness of addicts to infections may be partially due to opiate effects on immune cells. We find that acute morphine inhibits phagocytosis in murine peritoneal macrophages in vitro with apparent desensitization at high concentrations, whereas chronic exposure results in a state akin to tolerance/dependence where macrophages seem to require morphine to phagocytize at a control level. However, both putative desensitization and tolerance/dependence are reversible, since drug re-addition several hours after withdrawal results in inhibition, as in acute exposure. This shows that opiate effects on immune cells are variably related to the experimental context in which they are administered, which is of relevance for understanding their potential role in immunosuppression. PMID- 10669068 TI - Reductions in acquisitive crime and drug use after treatment of addiction problems: 1-year follow-up outcomes. AB - The relationship between acquisitive crime and drug misuse problems was studied among 753 clients recruited to the National Treatment Outcome Research Study (NTORS). More than 17000 offences were reported during the 90-day period prior to treatment. Half of the clients committed no acquisitive crimes during this period whereas 10% committed 76% of the crimes. At 1-year follow-up, the number of crimes was reduced to one third of intake levels, and criminal involvement was reduced by about half. Reductions in regular heroin use were strongly associated with reductions in crime. The reduction in crime following treatment is of great importance and provides immediate benefit to society through the reduced economic costs of crime. PMID- 10669069 TI - The impact on retention of expansion of an Australian public methadone program. AB - Although rapid expansion of methadone programs has occurred in many countries, there are few studies of the impact on treatment success. The one public methadone maintenance program in Canberra was expanded from 85 places in 1991 to 350 places in late 1992. While this responded to a real need, it also led to a temporary increase in drop-outs. Retention was considerably improved in 1996 when clients began to be transferred from the public program to fee-for-service primary health care, but the improvement was also not sustained. In recent years, one-quarter to one-half of clients dropped out before stabilization. Multiple entries into treatment were common and overall retention was not affected by previous treatment episodes or length of time between episodes. Methadone programs should monitor retention as part of on-going evaluation and improvement of treatment policy. PMID- 10669070 TI - Substance use among high school students in Greece: outburst of illicit drug use in a society under change. AB - Trends in self-reported substance use in surveys of Greek adolescent students show that regular tobacco smoking increased in 1998 (20.8%) after having fallen between 1984 (22%) and 1993 (14.6%). Frequent alcohol consumption decreased in 1998 to 12.1%, from around 15% in both previous surveys. A sharp increase was observed in illicit drug use from 6% in 1993 to 13.7% in 1998. Unprescribed use of psychoactive medicines continued to decrease steadily from 53.8% in 1984 to 31.8% in 1998. Perceived availability and risks of cannabis parallel trends in use. Policy measures and recent sociocultural changes seem to influence illicit use. PMID- 10669071 TI - A comparison of the harms associated with the injection of heroin and amphetamines. AB - An investigation into whether or not the level of harm associated with injecting drug use varies depending on the drug that is injected was conducted among 151 primary heroin injectors and 145 primary amphetamine injectors. Compared to primary amphetamine injectors, primary heroin injectors were more dependent on their primary drug, had poorer social functioning, and had recently exhibited a higher degree of criminal behaviour. There were no differences between the two groups in terms of the prevalence of needle sharing, their health, or their psychological functioning, despite the amphetamine users being significantly younger and having used less frequently. It is concluded that while there are some harms that are attributable to injecting per se, the type of drug that is injected does play a mediating role in the relationship between injecting drug use and its associated harm. PMID- 10669072 TI - Illicit drugs and driving: prevalence, beliefs and accident involvement among a cohort of current out-of-treatment drug users. AB - Drug-driving behaviour among out-of-treatment dependent drug users has not been investigated while a theoretical perspective on the propensity of certain drug users to drive while impaired has not been suggested. This paper examines illicit drugs and driving behaviour and accident involvement among out-of-treatment current drug users. Psychological evidence of belief-based mechanisms to account for the decision to drive while impaired by drugs are provided. A total of 210 out-of-treatment current drug users were interviewed in a non-clinical setting by privileged access interviewers. Questionnaire measures were: current illicit drug use, severity of dependence, illicit drugs and driving behaviour, impaired and unimpaired accident involvement and beliefs and perceptions about the impairing effects of a number of illicit drugs. Analyses are restricted to participants who reported driving during the previous 12 months (n = 71). Fifty-eight participants (81.7%) reported driving immediately after consuming illicit drugs, primarily heroin and cannabis. Of these 41.4% (n = 24) had at least one road accident as a driver, 15 of whom (62.4%) reported accident involvement following recent drug consumption. Belief-based results showed that participants who reported never driving after using illicit drugs perceived heroin, methadone and alcohol to be greater significance for accident risk and driving skills impairment than other drugs. Those drivers who reported drugs and driving behaviour believed only alcohol to be significantly more impairing than other drugs. Findings indicated that illicit drugs and driving behaviour is common among out-of-treatment drug users. Accident involvement among this cohort is characterised by the previous consumption of illicit substances. Differential beliefs about the effects of drugs on driving performance and accident risk were shown to be dependent upon frequency of drugs and driving behaviour. Results are discussed in terms of experiential factors and consistency theories of attitude formation and change. PMID- 10669073 TI - The brief abstinence test: voucher-based reinforcement of cocaine abstinence. AB - This study assessed the effectiveness of a brief abstinence reinforcement procedure for initiating cocaine abstinence in methadone maintenance patients. On Monday of the test week, 72 cocaine-abusing methadone patients were offered a $100 voucher if urine samples collected on Wednesday indicated that they had abstained from cocaine across that 2-day period. A patient was considered abstinent and the voucher delivered if the urine benzoylecgonine concentration decreased by 50% from Monday to Wednesday (quantitative criterion) or if the concentration of Wednesday's urine sample was < or = 300 ng/ml. Overall, 79% of study patients showed urinalysis evidence of abstention from cocaine between Monday and Wednesday of the test week. In a subsample with complete data (n = 50), significantly more patients abstained from cocaine from Monday to Wednesday of the test week (84%) than from Monday to Wednesday of the week before (36%) or after (32%) the test week. Furthermore, while almost all patients (94%) decreased their benzoylecgonine concentration from Monday to Wednesday of the test week, significantly fewer patients' benzoylecgonine concentrations decreased from Monday to Wednesday of the week before (56%) or after (48%) the test week. This highly efficacious procedure may have clinical application where reliable abstinence initiation is desired, either on a temporary basis (e.g. sobriety sampling) or at the start of longer-term interventions. It may also be possible to use the brief abstinence test as an experimental model to assess the effects of other therapeutic interventions on abstinence initiation in treatment settings. PMID- 10669074 TI - Ultrastructure of human dentine 40 years ago--progress and perspectives. AB - In the 1959 premier issue of the Archives of Oral Biology, the first TEM observations were presented of sections of undecalcified human mature dentine produced with diamond knives. The odontoblast process was clearly shown to be a cytoplasmic extension of the odontoblast. The peritubular dentine appeared to be more calcified than the intertubular dentine and contained hydroxyapatite as demonstrated by selective area electron diffraction. In the 40 years which followed, significant progress was made in TEM methodology, including improvements in fixation and embedding, development of ultrastructural cytochemistry and immunocytochemistry, and the use of electron microscope autoradiography. Additionally, we saw the advent of SEM and HRTEM. Thus, better knowledge was gained of (1) the odontoblast and its process and the lamina limitans, (2) the dentinal nerve fibrils and (3) the HRTEM aspects of dentine. Interestingly, to the present day, diamond knives have continued to serve as the best tool for preparing thin sections of non-decalcified mature hard tissue for TEM and HRTEM, not only for dentine but also for bone, enamel and cementum. PMID- 10669075 TI - Review of fluoride research since 1959. AB - The evidence for the two main hypotheses proposed for the mode of action of F in reducing caries is reviewed. The current conclusion is that low concentrations of F in plaque, which need frequent renewal, favour remineralization of dental tissue (i.e. a net reduction of demineralization) in the later stages of a cariogenic episode. This replaces the other view that a high concentration of F in the tooth mineral reduces its solubility. PMID- 10669076 TI - Progress in oral biology research, 1959-1999. A review and update of Volume 1. PMID- 10669077 TI - A comparison of four extraction methods for substance P, neurokinin A and calcitonin gene-related peptide from human dental pulp tissue. AB - Measuring neuropeptides in biological tissues by radioimmunoassay requires efficient extraction that maintains their immunoreactivity. Many different methods for extraction have been described, but there is little information on optimal extraction methods for individual neuropeptides from human dental pulp tissue. The aim was therefore to identify an effective extraction procedure for three pulpal neuropeptides; substance P, neurokinin A and calcitonin gene-related peptide. Tissue was obtained from 20 pulps taken from teeth freshly extracted for orthodontic reasons. The pulp samples were divided into four equal groups and different extraction methods were used for each group. Boiling whole pulp in acetic acid gave the highest overall yield and, in addition, offered an easy and rapid means of pulp tissue processing. The use of protease inhibitors did not increase the recovery of the immunoreactive neuropeptides but did provide the best combination of maximal recoveries and minimal variability. These results should be useful for planning the extraction of these neuropeptides from human pulp tissue in future studies. PMID- 10669078 TI - Variations in human masseter and temporalis muscle activity related to food texture during free and side-imposed mastication. AB - Adjustments of mastication to food texture have been examined in various studies, but the notion of food texture is often ill defined and usually assessed in terms of hardness. The goal of this study was to examine the pattern of activity in masseter and temporalis muscles during mastication of different food samples with known textural properties and to determine the interindividual variability. Electromyograms were recorded from the right and left masseter and temporalis muscles in 36 young adults during 'free-style' and side-imposed mastication. Five different types of food with known rheological properties were used. Both temporalis and masseter activity increased with increased stress at maximum strain of the chewed samples. A power function optimally described the relation between muscle work per chew and the mechanical measurements of food; this confirmed that the masticatory process is adjusted to accommodate to food texture. Temporalis muscle activity was more influenced by food texture than was masseter muscle activity. Less muscle work was needed to prepare the food bolus for swallowing during free-style mastication. However, 25% of the participants showed no differences between unilateral side-imposed mastication and 'free style', suggesting that they might have greater chewing efficiency on one side. PMID- 10669079 TI - Effects of dietary consistency and water content on parotid amylase secretion and gastric starch digestion in rats. AB - The aim was to estimate the significance of oral sensation and mastication in inducing amylase secretion from the parotid gland and subsequent starch digestion in the stomach. Rats were fed three diets of similar chemical composition but different physical presentations. Two were solid, either pellets or powder, and one was liquid. Oral sensory activity would be greatest with the pellets and least with the liquid. Only the pellets would require significant mastication. Three criteria were used to estimate amylase secretion, amylase activity in the stomach, the depletion of glandular amylase activity and plasma amylase concentrations. Gastric starch digestion was estimated by measuring the concentration of reducing-sugars in the stomach contents. Parotid amylase secretion and gastric starch digestion were similar whether rats were fed pelleted or powdered solid food but much lower in rats fed a liquid diet. These findings support the view that it is the contact of dry food with the oral mucosa rather than the jaw movements involved in mastication that stimulates parotid amylase secretion. PMID- 10669080 TI - Viscoelastic properties of canine temporomandibular joint disc in compressive load-relaxation. AB - The present study was designed to investigate the mechanical properties and load relaxation of these discs in compression. Eight discs were used for the experiments. Compression was applied to specimens up to the specified strain, and a series of load-relaxation tests was conducted on each specimen from 0.25% strain to 2.0% strain with 0.25% intervals. The load-relaxation was monitored over a period of 2 min. The elastic moduli of the canine articular discs were 30.9 and 15.8 MPa at t = 0 and 120 sec, respectively, and the discs exhibited near-linear elastic characteristics at each time within a 2-min period. At all strains, the time-dependent load-relaxation curves showed that the load decreased markedly for the initial 30 sec, after which it levelled off after 120 sec with a steady nonzero level. This relaxation feature can be well represented by Kelvin's model. It is concluded that the canine temporomandibular joint disc can be represented as a linear viscoelastic material, and that it plays an important part as a stress absorber under compression. PMID- 10669081 TI - Localization of plasminogen activators and plasminogen-activator inhibitors in human gingival tissues demonstrated by immunohistochemistry and in situ hybridization. AB - The plasminogen-activating system plays an important part in tissue proteolysis in physiological as well as pathological processes. Plasminogen activators u-PA (urokinase) and t-PA (tissue) as well as the inhibitors PAI-1 and PAI-2 are present in gingival crevicular fluid in concentrations significantly greater than in plasma. This fact, and the finding that the concentrations of t-PA and PAI-2 are higher in areas with gingival inflammation, indicate local production of these components. The present study describes, by means of in situ hybridization and immunohistochemistry, the localization of the plasminogen activators and their inhibitors in gingival tissues from patients undergoing periodontal surgery. t-PA mRNA and t-PA antigen were primarily found in the epithelial tissues, predominantly in the sulcular and junctional regions, although occasionally in the oral epithelium and in blood vessels of the connective tissue. u-PA and u-PA-receptor signals were seen in single cells within the junctional and sulcular epithelia and adjacent to blood vessels close to the junctional epithelium, but rarely in the oral epithelium. Similar to t-PA, the predominant location of PAI-2 mRNA was the gingival epithelia. In the junctional and sulcular epithelia, PAI-2 mRNA was seen throughout the thickness, while in the oral epithelium the strongest signals were seen in stratum granulosum and stratum spinosum. PAI-1 mRNA was invariably found in the connective tissue associated with blood vessels. The present study confirms earlier indications of local production of plasminogen activators and their inhibitors in gingival tissues. In addition, the results demonstrate that t-PA and PAI-2 in these patients are produced predominantly in the epithelial tissues. Furthermore, the presence of t-PA and PAI-2 seems to be most pronounced in the areas likely to be subjected to bacterial assault. PMID- 10669082 TI - Innervation and myoepithelial arrangements in the submandibular salivary gland of ferret investigated by enzyme, catecholamine and filament histochemistry. AB - Although the submandibular gland of ferret is useful for studying salivary secretory processes which are regulated by nerves and involve myoepithelial activity, little attention has been paid to its parenchymal innervation and myoepithelial arrangements. Therefore, glands obtained postmortem from mature ferrets of both sexes were here examined with the use of light-microscopic histochemical techniques for cholinesterases, phosphatases and phosphorylase, histofluorescence for catecholamines, and milling dyes. Acetylcholinesterase staining was associated with nerve trunks in the interlobular stroma and an extensive intralobular network of nerve fibres, presumably of a cholinergic type, embracing acini and ducts. There were fewer fibres containing fluorescing catecholamines, presumably adrenergic. They were largely associated with acini. Numerous stellate cells with fine branching processes embracing acini, presumably myoepithelial cells, and a few spindle-shaped basal cells, investing striated ducts, were demonstrated on frozen tissue by alkaline phosphatase, but not by adenosine triphosphatase, inosine diphosphatase and phosphorylase. Cells of similar shape and distribution were also demonstrated by staining with milling dyes on fixed tissues, indicating possibly a filamentous constituent conferring mechanical stability and/or contractile ability. Together, these results suggest, firstly, that a cholinergic-type parenchymal innervation is prominent in the submandibular gland of ferret, although many adrenergic nerves are also present, and, secondly that the gland has a very extensive myoepithelial network which is possibly involved in membrane transport, and the support and or contraction of the secretory parenchyma. PMID- 10669083 TI - T-cell antigen specificity in humans following stimulation with Porphyromonas gingivalis. AB - The effects of Porphyromonas gingivalis stimulation on T-cell clonality and cytokine mRNA expression in peripheral blood mononuclear cells from individuals with gingivitis and periodontitis were investigated. Clonality of T cells was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and single-strand conformation polymorphism analysis. Cytokine mRNA expression was investigated by RT-PCR. Whereas unstimulated mononuclear cells did not demonstrate obvious clonality, clonal expansion was found in most Vbeta families after stimulation. However, there was no relation between clonal change and disease category or the presence of P. gingivalis infection. Messenger RNA for interferon-gamma and interleukin-13 was upregulated whereas interleukin-4 and -10 were downregulated following P. gingivalis stimulation. Interleukin-12p35 demonstrated no consistent pattern. This study supports the concept that P. gingivalis stimulates T cells in an antigen-specific fashion. It further suggests that peripheral blood T cells may preferentially produce interferon-gamma and interleukin-13 in response to P. gingivalis stimulation irrespective of disease or P. gingivalis status. PMID- 10669084 TI - Inhibition of rat parotid ecto-ATPase activity. AB - The inhibitory profile of several known and suspected ecto-ATPase inhibitors was compared on ecto-ATPase activity in rat parotid plasma membranes. Those chemicals with high IC50 (above 130 microM) were the nucleotides alpha,beta-methylene ATP, beta,gamma-methylene ATP, 2-methylthio ATP, inosine triphosphate, 5'-p fluorosulphonylbenzoyladenosine, the sulphonates, 1-amino-2-naphthol-4-sulphonic acid, Coomassie brilliant blue G, and the stilbene disulphonates, DIDS and SITS. Those agents with low IC50 were: Coomassie brilliant blue R (114 microM), ATPgammaS (49 microM), suramin (72 microM) and Reactive blue 2 (28 microM). The last three inhibitors have similar potencies as inhibitors of ATP hydrolysis by whole parotid acinar cells. ARL67156, a selective inhibitor of ecto-ATPase, had an IC50 of approx. 120 microM. Suramin displayed non-competitive inhibition of ecto-ATPase whereas the inhibitory effects of ATPgammaS and Reactive blue 2 were curvilinear on Dixon plots. These results define the effects of various agents on ecto-ATPase in an exocrine tissue that has been shown to respond to extracellular ATP. PMID- 10669085 TI - Tenacious adhesion of Actinobacillus actinomycetemcomitans strain CU1000 to salivary-coated hydroxyapatite. AB - Adherence of Actinobacillus actinomycetemcomitans to hard-tissue surfaces was evaluated by comparing a phenotypically stable, well-maintained clinical isolate (strain CU1000) to Streptococcus gordonii G9B, an extensively studied oral colonizing bacterium. Standard innocula of radiolabelled bacteria were added to saliva-coated hydroxyapatite (SHA) and the ratio of bound to unbound cells counted. Several other clinical isolates as well as laboratory strain Y4 were studied. In other experiments, cell detachment from SHA was compared in static and shaking vessels to calculate controlled desorption of cells over time. A sonic-displacement assay was used to measure avidity of binding to HA and SHA. To better define the attachment properties of CU1000, bacteria were treated with a variety of agents including detergents, salts and enzymes before or after incubation with SHA. Results indicated that CU1000 bound better than G9B (a minimum of 10-fold greater; p < or = 0.05) and did not desorb from SHA, while G9B desorbed to equilibrium in 4 h. Furthermore, Langmuir isotherm calculations indicated that, unlike G9B, CU1000 did not follow second-order adsorption kinetics and thus did not achieve saturation. In addition, of the agents tested only periodate reduced attachment and resulted in detachment of CU1000 from surfaces. These experiments suggest that clinical isolates of A. actinomycetemcomitans possess unique binding properties that promote adsorption to and impede desorption from SHA. The characteristics described for the actinobacillus in this study have been previously underestimated, appear to be mediated by glycoconjugates, and may resemble attachment described for several biofilm-forming, non-oral pathogens. PMID- 10669086 TI - The craniofacial complex in 45,X/46,XX females. AB - Fourteen Finnish 45,X/46,XX females were compared with population female and male controls, and in addition, nine of them were compared with their first-degree female relatives. Linear and angular measurements were made from standardized lateral cephalograms of patients and normal population controls from the "Kvantti" study series. In both comparisons the results indicated that craniofacial dimensions in 45,X/46,XX females were smaller than those in population female and male controls. The general metric pattern was similar to that observed in relation to the tooth crowns of 45,X/46,XX females. Several of their craniofacial proportions and plane angles differed from those of normal women: shorter anterior and posterior cranial bases and a flatter cranial-base angle, a sagittally shorter maxilla and a sagittally shorter mandible with an enlarged ramus:corpus length ratio, posterior rotation of the mandible and a tendency to bimaxillary retrusion. It is suggested that the reduction of X chromosomal genetic material in 45,X/46,XX females results in smaller craniofacial dimensions than in normal females, with substantial effects on dimensional ratios and especially plane angles of the cranial base. PMID- 10669087 TI - The buffer capacity and buffer systems of human whole saliva measured without loss of CO2. AB - The buffer capacity of unstimulated (UWS) and stimulated (SWS) whole-mouth saliva involves three major buffer systems. The aim was to determine the buffer capacity of UWS and SWS at specific pH in the interval from pH 7.5 down to pH 3.0. The contribution of each of the buffer systems was also determined under conditions resembling those in the mouth. UWS and SWS were collected from 20 healthy volunteers; the saliva was collected under paraffin oil in order to avoid loss of CO2. The buffer capacity of UWS and SWS in samples with and without bicarbonate (HCO3-) and CO2 were measured at various pH by acid titration in a closed system at 36 C. The mean concentrations of the buffer systems in UWS (mean flow rate 0.55 ml/min) were 4.4 mmol/l HCO3-, 4.5 mmol/l phosphate (of which 1.3 mmol/l was present in the form of HPO4(2-)), 1876 microg/ml protein; the saliva pH was 6.8 and the P(CO2) 29.3 mmHg. The corresponding mean concentrations in SWS (mean flow rate 1.66 ml/min) were 9.7 mmol/l HCO3-, 3.8 mmol/l phosphate (of which 1.9 mmol/l was present in the form of HPO4(2-)), 1955 microg/ml protein; pH 7.2 and P(CO2) 25.7 mmHg, The highest buffer capacity of UWS and SWS was 6.0 and 8.5 mmol H+ /(1 saliva*pH unit) at pH 6.25, respectively. At saliva pH in the range from pH 7 down to pH 5, the following had significant impact on buffer capacity: the HCO3- concentration (p < 0.001), the flow rate (p < 0.01), and the pH of the saliva (p < 0.05). At acidic pH in the range from pH 5 down to pH 4, however, only the protein concentration had a significant impact on buffer capacity (p < 0.01). PMID- 10669088 TI - Age-related changes in the D-aspartic acid content of the teeth of the senescence accelerated mouse. AB - It is known that D-aspartic acid increases with age in dentine. Here, age-related changes in the D to L-aspartic acid (D/L) ratios of the lower teeth of two different sublines of the senescence-accelerated mouse (SAM), SAMP2/Iw (SAM, prone 2/Iwate) and SAMR1/Iw (SAM, resistant 1/Iwate) were measured by gas chromatography. The D/L ratio of the molars increased with advancing age, whereas that of the incisors did not. In mice younger than 6 months of age the D/L ratio of the molars from SAMP2/Iw tended to be higher than that from SAMR1/Iw, whereas the converse applied to older mice. Racemization in the molars occurred significantly faster in SAMR1/Iw than SAMP2/Iw (p = 0.01-0.001). Analysis according to the kind of tooth showed that the D/L ratio increased gradually in the order incisors < third molars < second molars < first molars, indicating that the ratio was higher the earlier the molars formed. As racemization depends upon the environmental temperature, the rectal temperatures of the mice were also examined. The rectal temperature of SAMP2/Iw was highest when they were 2 months old, but declined rapidly thereafter, whereas the rectal temperature of SAMRI/Iw was highest when they were 6 months old, after which it declined gradually. These results indicate that the D-aspartic acid contents of the molars of SAMR1/Iw and SAMP2/Iw increase with age in a different fashion and suggest that the fashion was determined by the body temperature, but not by the senescence-accelerated age. PMID- 10669089 TI - Thrombospondin-2 gene expression and protein localization during embryonic mouse palate development. AB - The mammalian palate develops from projections of the paired maxillary processes termed palatal shelves. Shelf growth is an essential in normal palatal morphogenesis. Mesenchymal proliferation in the palatal shelves is modulated by transforming growth factor-1 (TGF-1), among other growth factors. Several pathways effect TGF-beta activation, including one which utilizes thrombospondin (TSP). TSP-1 is a major activator of TGF-beta in vivo and has been localized in head mesenchyme, including palates. TSP-2 appears to inhibit TSP-1 activation of latent TGF-beta by competitively binding the latent TGF-beta. Here the TSP-2 mRNA transcript and the immunolocalization of TSP-2 protein with progressive palatogenesis were quantified. There was a significant (p < 0.05) decline of TSP 2 transcript with palatal maturation; there was no evidence correlating the TSP-2 transcription with the amount of activated TGF-beta. At the vertical shelf stage of palatogenesis, TSP-2 protein was found throughout the extracellular matrix of shelf mesenchyme. By the horizontal shelf stage, TSP-2 protein was principally localized to the ossification centres of the developing maxilla, both in extracellular matrix and bone; far less was seen in palatal shelves proper. These results suggest that TSP-2 is multifunctional during embryonic palate formation. PMID- 10669090 TI - Development of multi-species consortia biofilms of oral bacteria as an enamel and root caries model system. AB - The aim was to establish defined-species consortium plaque biofilms to investigate enamel and root caries in an artificial mouth. Strains of the putative enamel and root caries pathogens, Streptococcus mutans, Strep. sobrinus, Actinomyces naeslundii and Lactobacillus rhamnosus, were screened in batch culture for potential cariogenic properties: a low terminal pH, ability to aggregate, and catabolic diversity. The strains selected were grown as monoculture biofilms and as consortium plaque biofilms in a multiplaque artificial mouth. The biofilms were supplied with a constant flow of a simulated oral fluid and were given periodic sucrose (and in some instances glucose) to simulate meals. All the bacteria except L. rhamnosus formed large, monospecies biofilms with resting pH in the range 5.3-5.8. The consortia biofilms were larger and had a resting pH of 4.9-5.3. The consortia biofilms supplied with 8-hourly carbohydrate comprised mainly 'mutans' streptococci (58, SD 5.5%) and L. rhamnosus (42, SD 5.7%). A. naeslundii characteristically was absent or present in a low percentage (up to 4% colony-forming units). All biofilms demineralized polished bovine enamel and dentine blocks, as assessed by microradiography and enamel-surface microhardness measurement. The consortia also demineralized intact enamel and tooth roots; they were more cariogenic to enamel than any of the monoculture biofilms, as measured by enamel-surface softening, but variation in lesion depth was proportional to biofilm wet weight irrespective of acidogen composition (r = 0.93, p < 0.05). Enamel lesions had a well-mineralized intact surface and a zone of subsurface demineralization, typical of early natural lesions. Dentine and root lesions showed extensive demineralization but lacked a pronounced surface mineralized zone. Substitution of glucose for sucrose had no effect on the cariogenicity of the consortium to bovine enamel or human roots and had no major effect on the plaque composition. Continuously supplied fluoride (19 parts/10(6)) resulted in a substantially reduced enamel surface softening and subsurface demineralization of intact roots. It was concluded that consortia biofilms of selected caries pathogens generate realistic caries lesions in all tooth hard tissues under controlled growth conditions in the artificial mouth. This in vitro caries experimental model may prove useful for the study of interrelations between the plaque biofilm, tooth tissues and the oral environment, and for the development of procedures to modify the course of caries development. PMID- 10669091 TI - The molecular cloning, nucleotide sequence and expression of an antigenic determinant from Porphyromonas gingivalis. AB - A genomic library generated in Escherichia coli was probed with a monoclonal antibody (mAb) LDS28, which reacts with a species-specific cell-surface antigen of Porphyromonas gingivalis. A clone designated pGPR2.1 was shown to express a 46 kDa protein reactive with mAb LDS28, which maps to a 1.7-kb HincII fragment. DNA sequence analysis revealed pGPR2.1 contains a 5653-bp insert with six open reading frames, one of which shows significant DNA homology with the rnhB gene of E. coli. Several subclones of pGPR2.1 were randomly generated in plasmid vector pTTQ18* using restriction enzyme Sau3a. Immunoblotting of subclones demonstrated that the LDS28-reactive antigen was coded for by an open reading frame predicted to specify a protein of 455 amino acids (50 kDa). This open reading frame was designated pgaA (Porphyromonas gingivalis antigen). The predicted amino acid sequence of PgaA contains a putative ABC signature for binding NTPs as well as a predicted transmembrane domain. Minicell labelling of pGPR2.1-encoded proteins and subclone derivatives revealed that pgaA directs expression of protein of multiple molecular weights (31-46 kDa) from its own promoter in E. coli, and that some of these forms may be caused by proteolysis of a 50-kDa precursor which itself shows a reduced apparent molecular weight (46 kDa) on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. PMID- 10669092 TI - Increased blood flow and nerve firing in the cat canine tooth in response to stimulation of the second premolar pulp. AB - Mustard oil or mechanical stimulation was applied to maxillary second premolar tooth pulps and pulpal blood flow and or intradental nerve activity in the ipsilateral canine tooth were recorded in the cat. Mustard oil application to the second premolar pulp significantly increased blood flow in the canine tooth pulp to 162.0+/-65.8% (n = 16) of the prestimulation flow compared to control data obtained with application of mineral oil (107.0+/-5.1%, n = 6) (Mann-Whitney U test, p = 0.0009). Sectioning of the infraorbital nerve and its branches on the experimental side (n = 4) did not affect this increase in pulpal blood flow. The paraperiosteal injection of 2% lidocaine (1.0 ml) without vasoconstrictor significantly inhibited the increase in canine pulpal blood flow induced by mustard oil application to the second premolar pulp (109.8+/-6.8% of the prestimulation level, n = 7) (Mann-Whitney U-test, p = 0.0013). Sporadic firing or sometimes bursts of action potentials in the canine pulp nerves were recorded during and/or after the mustard oil application to the second premolar pulp in three of 16 cases. Four single pulp nerve units firing in synchrony with the mechanical stimulation of the second premolar pulp were recorded in two of eight canines, which substantiated the existence of branched afferents innervating both teeth. These findings suggest that stimulation of the second premolar pulp may induce axon reflex-related vasodilation and intradental nerve firing in the canine pulp via branched afferent fibres innervating both the second premolar and canine teeth. PMID- 10669093 TI - Expression of cathepsin K mRNA during experimental tooth movement in rat as revealed by in situ hybridization. AB - The expression of cathepsin K. a novel collagenolytic enzyme specifically expressed in osteoclasts, was investigated in the rat maxillary dentoalveolar unit during experimental tooth movement by in situ hybridization histochemistry with a non-radioisotopic cRNA probe for rat cathepsin K. Orthodontic elastics were inserted into the interproximal space between the maxillary first and second molars of 7-week-old male SD rats according to Waldo's method and sections prepared from tissues obtained at 12 hr, 1, 2, 3, 4, 7, and 12 days after orthodontic force application. Cathepsin K mRNA expression was detected in the mono- and multinuclear osteoclasts on the pressure side of the alveolar bone at 12 hr after force application, and the distribution and number of cathepsin K mRNA-positive osteoclasts increased time-dependently on the pressure side. At 3-4 days, a marked increase in cathepsin K mRNA-positive osteoclasts was found not only on the pressure side but also on the tension side of the alveolar bone in response to tooth movement. At 7-12 days, the cathepsin K mRNA-positive osteoclasts on both sides had disappeared. These findings suggest that the recruitment of osteoclasts on the pressure side begins during the initial stage of orthodontic tooth movement and the site-specific early induction of cathepsin K mRNA may cause an imbalance in the relative resorption activities on the pressure and tension side incident to such movement. PMID- 10669094 TI - Expression of myogenic regulatory factors during the development of mouse tongue striated muscle. AB - While the role of myogenic regulatory factors (MRFs) in skeletal myogenesis has been well evaluated in limb and trunk muscles, very little is known about their role in tongue myogenesis. Here the expression of MRF mRNA in mouse tongue muscle was examined during development from embryonic day (E)11 to birth and compared them with that in hind-limb muscle. Desmin, muscle creatine kinase and troponin C mRNAs were used as markers for myoblast determination, myotubule formation and myofibre maturation, respectively. The mRNA quantities were determined by competitive reverse transcriptase-polymerase chain reaction. The expression profile of desmin mRNA indicated that myoblast determination occurred before E11 in both the tongue and hind-limb muscles; the profile of muscle creatine kinase and troponin C mRNAs indicated that myotubule formation and myofibre maturation began between E11 and 13 in both tongue and hind-limb muscles, but ended 2 days earlier in the tongue than in the hind limb. Expression of myoD and myogenin mRNAs began at E11, increased, and showed peak values earlier in the tongue muscle (E13) than in the hind-limb muscle (E15). Expression of MRF4 mRNA appeared earlier in the tongue (E13) than in the hind-limb muscle (E15) and increased in both muscles after that. These results suggest that myotubule formation and myofibre maturation in the tongue muscle progress faster than in the hind-limb muscle, a result of earlier expression of myoD, myogenin, and MRF4 in response to earlier functional demands such as suckling immediately after birth. PMID- 10669095 TI - Mutational analysis of X-linked amelogenesis imperfecta in multiple families. AB - Seven mutations in the amelogenin gene are associated with X-linked amelogenesis imperfecta. These mutations can produce reductions in the amount of enamel and the degree of mineralization. Two families have been identified from western North Carolina exhibiting features of amelogenesis imperfecta, characterized by brown enamel in affected males and interposed vertical bands of normal appearing and brown enamel in presumably heterozygous females. Mutational analysis reveals a C-A mutation in exon 6 at codon 41 of the X-chromosomal amelogenin gene, resulting in a pro-thr change in all individuals having the amelogenesis imperfecta phenotype. This mutation was previously reported in a family with X linked hypomaturation amelogenesis imperfecta. There is no known relationship between any of the three families but the presence of similar phenotypes and common mutations suggests they may be distantly related. For individuals from all three families, the haplotype for six highly polymorphic loci flanking the amelogenin gene was determined. A common haplotype was demonstrated among two of the three families, suggesting that the mutation may have been inherited from a common ancestor. The finding that the third family had a distinct haplotype may indicate that the C-A mutation at codon 41 represents a mutational hotspot that occurs with greater frequency than other known amelogenin gene mutations. The phenotype resulting from this mutation was highly consistent in affected male members of the same family and between families. PMID- 10669096 TI - The Twirler mouse, a model for the study of cleft lip and palate. AB - Twirler (Tw) is a semidominant mutation in the mouse affecting the embryonic development of the midfacial region. Most heterozygous Tw mice, +/-, become obese at adulthood with a concomitant decrease in fertility. Homozygous mice have clefts of the midfacial region and a disrupted nasal cavity. Midfacial clefts included clefts of the palate combined with either unilateral or bilateral clefts of the lip. The clefts of the lip were either complete or incomplete. The palatal shelves in Tw/Tw were very much reduced. Apart from these defects, homozygous Tw looked normal, and were born alive, although they reportedly die within 24 h after birth. It is proposed that the Twirler model can be used to improve understanding of the genetic mechanisms involved in the normal development of the midfacial region. PMID- 10669097 TI - Cryptosporidiosis in birds--a review. AB - The morphology, life cycle, maintenance, host specificity, incidence of Cryptosporidium species infecting birds, as well as the epidemiology, clinical signs, pathology, immunology, diagnosis, therapy, and control of avian cryptosporidiosis are reviewed. Based on the accepted criteria used for differentiation of Cryptosporidium isolates into valid species, this review places the validity of C. meleagridis in doubt and suggests that C. meleagridis isolated from birds is very closely related to, or identical with C. parvum infecting more than 100 species of mammals. PMID- 10669098 TI - Rate of infection and of reinfection by Echinococcus granulosus in rural dogs of the province of Rio Negro, Argentina. AB - Hydatidosis (cystic echinococcosis) constitutes a serious public health problem in the Province of Rio Negro, Argentina. In the present work it was intended to evaluate the prevalence of the canine echinococcosis in rural areas of the Province of Rio Negro and studied the reinfection rate in dogs after treatment with Praziquantel during the period 1980-1997. A total of 496 dogs were studied in 18 canine concentrations in order to establish the initial prevalence rate which was 42.3%. From 1980 onward dogs should have been systematically treated with anthelmintic every 2 months in rural areas and every 6 months in urban areas. We estimated that approximately 65% of dogs were treated. To determine the reinfection rate, 476 dogs (1984) and 598 dogs (1996) were studied after anthelmintic treatment during two sequential time periods (0-45; 46-90 days). In both cases treated animals were compared with untreated dogs. Prevalences were 3.5%, 6.7% and 21.3% in 1984 and 0.8%, 4.0% and 10.0% in 1996. For the purpose of surveillance a total number of 21,444 dogs were studied during 18 years. Prevalence of Echinococcus granulosus decreased significantly in the first year from 42.3% to 6.1%. PMID- 10669099 TI - Dynamics of the humoral immune response of calves infected and re-infected with Cooperia punctata. AB - The dynamics of the humoral immune response of calves were analysed after primary infection and re-infection with the intestinal nematode Cooperia punctata. 12 male 5 month-old Holstein-Friesian calves were randomly divided into two groups A and B. At the beginning of the experiment Group A animals were each infected experimentally with a single oral dose of 130,000 infective third stage larvae (L3) of C. punctata. The animals of Group B were kept as non-infected controls. The two calves from Group A with the highest infections died of cooperiosis at 32 and 44 days after infection (DAI), respectively. On DAI 100 the calves were treated with the recommended dose of oxfendazole. On DAI 180 the remaining four calves of Group A and three animals of Group B (B1) were infected with 260,000 L3 of C. punctata, while the other three calves of Group B (B2) served as non infected controls. Monitoring of the humoral immune response predominantly demonstrated an IgG1 response against both adult and L3 antigen of C. punctata. Moreover, re-infections increased the levels of these immunoglobulins. IgA levels were less increased than IgG1 and no significant increase was observed in IgG2 and IgM levels. Immunoblotting analysis showed that total IgG present in the serum of the primary infected animals mainly reacted against adult proteins of 12 14 and 17-20 kDa and against L3 proteins of 33 and 43 kDa. After re-infection total IgG reacted with the same adult proteins but also with an adult 29 kDa protein. PMID- 10669100 TI - Establishment and characterization of an Oklahoma isolate of Anaplasma marginale in cultured Ixodes scapularis cells. AB - Anaplasma marginale is a tick-borne hemoparasite of cattle worldwide. The Virginia isolate of A. marginale was propagated previously in a cell line derived from embryos of the tick, Ixodes scapularis. The cultured Anaplasma (VA-tc) was passaged continuously for over 4 years and retained its infectivity for cattle and antigenic stability. We report herein the continuous in vitro cultivation of a second isolate of A. marginale derived from a naturally infected cow in Oklahoma (OK-tc). Blood from the infected cow was subinoculated into a splenectomized calf and blood collected at peak parasitemia was frozen, thawed and used as inoculum on confluent tick cell monolayers. Colonies of Anaplasma were apparent in low numbers at 9 days post exposure (PE) and infection in monolayers reached 100% by 4-5 weeks PE. Cultures were passaged by placing supernatant onto fresh tick cell monolayers at a dilution of 1:5 or 1:10. By the third passage development of the OK-tc was similar to that of the VA-tc and a 1:5 dilution resulted in 100% infection in 10-12 days. Inoculation of OK-tc into a splenectomized calf caused clinical anaplasmosis and Dermacentor ticks that fed on this calf transmitted the organism to a second susceptible calf. Major surface proteins (MSPs) 1-5 of the OK-tc were compared with homologous proteins present on VA-tc and the erythrocytic stage of the Oklahoma isolate. The MSPs 1, 2, 4, 5 were conserved on the OK-tc but there was evidence for structural variation in MSP3 between the cultured and erythrocytic stage of Anaplasma. MSP2 and MSP3 were the major proteins recognized by serum from infected cattle. Two-dimensional gels also identified positional differences between VA-tc and OK-tc in MSP2 and MSP3. The OK-tc may have potential to be used as antigen for development of an improved vaccine for anaplasmosis in the South Central United States. PMID- 10669101 TI - A model to simulate the effect of vaccination against Boophilus ticks on cattle. AB - This paper describes a vaccination model to simulate the effect of cattle vaccination with concealed antigens on Boophilus tick spp. The model considers the vaccination effect in three parts: antibody titer, accumulation of damaging vaccination effects by parasite stages, and the effect of accumulated damage on all tick life stages. Biological parameters for ticks and hosts, as well as parameters describing tick-host interaction, were included. The validity of this model, integrated with the TICKSIM simulation program, was demonstrated for the Bm86-containing vaccine Gavac by comparing simulated and real data for several geographic locations in the Americas. All model parameters were estimated using field data collected in the different geographic locations. The model sensitivity to changes in antibody titer level and titer half-life was studied, and the impact on tick population density of changes in these parameters was evaluated. Simulation results showed that to achieve a higher level of tick control, an increase in the maximum antibody titer levels was more important than extending titer half-life in geographical locations with short seasonal peaks of tick infestation. The TICKSIM program, integrated with the new vaccination model, proved to be a framework for designing and evaluating tick control strategies, including vaccination with GavacTM. PMID- 10669102 TI - Comparative distribution of ivermectin and doramectin to parasite location tissues in cattle. AB - Pharmacokinetic studies have been used traditionally to characterize drug concentration profiles achieved in the bloodstream. However, endectocide molecules exert their persistent and broad spectrum activity against parasites localized in many different tissues. The aim of this study was to compare the distribution of ivermectin (IVM) and doramectin (DRM) to different tissues in which parasites are found following subcutaneous administration to calves. Holstein calves weighing 120-140 kg were injected in the shoulder area with commercially available formulations of IVM (Ivomec 1% MSD AGVET, NJ, USA) (Group A) or DRM (Dectomax 1%, Pfizer, NY, USA) (Group B). Two treated calves were sacrificed at 1, 4, 8, 18, 28, 38, 48 or 58 days post-treatment. Plasma, abomasal and small intestinal fluids and mucosal tissues, bile, faeces, lung and skin samples were collected, extracted, derivatized and analyzed by high performance liquid chromatography (HPLC) with fluorescence detection to determine IVM and DRM concentrations. IVM and DRM were distributed to all the tissues and fluids analyzed. Concentrations >0.1 ng/ml (ng/g) were detected between 1 and 48 days post-treatment in all the tissues and fluids investigated. At 58 days post treatment, IVM and DRM were detected only in bile and faeces, where large concentrations were excreted. Delayed Tmax values for DRM (4 days post administration) compared to those for IVM (1 day) were observed in the different tissues and fluids. High IVM and DRM concentrations were measured in the most important target tissues, including skin. The highest IVM and DRM concentrations were measured in abomasal mucosa and lung tissue. Enhanced availabilities of both IVM (between 45 and 244%) and DRM (20-147%) were obtained in tissues compared to plasma. There was good correlation between concentration profiles of both compounds in plasma and target tissues (mucosal tissue, skin, and lung). Drug concentrations in target tissues remained above 1 ng/g for either 18 (IVM) or 38 (DRM) days post-treatment. The characterization of tissue distribution patterns contributes to our understanding of the basis for the broad-spectrum endectocide activity of avermectin-type compounds. PMID- 10669103 TI - Claudin-1 contributes to the epithelial barrier function in MDCK cells. AB - Tight junctions (TJs) create a paracellular permeability barrier and also act as a fence preventing intermixing of proteins and lipids between the apical and basolateral plasma membranes. Recently, claudin-1 has been identified as an integral membrane protein localizing at TJs, and introduced claudin-1 can form TJ like networks in fibroblasts. To investigate the function of claudin-1, MDCK cells were transfected with a mammalian expression vector containing myc-tagged mouse claudin-1, and four stable clones were obtained. The myc-tagged claudin-1 precisely colocalized with both occludin and ZO-1 at cell-cell contact sites, indicating that exogenous claudin-1 was properly targeted to the TJs. Immunoblot analysis revealed that overexpression of claudin-1 increased expression of ZO-1 but not of occludin or ZO-2. The barrier functions of these cells were evaluated by transepithelial electrical resistance (TER) and paracellular flux. Claudin-1 expressing cells exhibited about four times higher TER than wild-type MDCK cells. Consistent with the increase of TER, the cells overexpressing claudin-1 showed reduced paracellular flux, estimated at 4 and 40 kD FITC-dextrans. These results suggest that claudin-1 is involved in the barrier function at TJs. PMID- 10669104 TI - Dynamics of gap junctions observed in living cells with connexin43-GFP chimeric protein. AB - To study the aggregation of cell-to-cell channels into gap junctions at individual cell-cell contacts, we transfected cells with an expression vector for a chimeric protein composed of the cell-to-cell channel protein connexin43 and a green fluorescent protein. The chimeric channel protein was visualized in the fluorescence microscope and was found to form gap junctions at the cell-cell contacts just like wild-type connexin43. Cells expressing the chimeric protein had functional cell-to-cell channels. Using timelapse videomicroscopy on live cells we observed individual gap junctions over long periods and recorded the time course of aggregation of the chimeric channel protein into gap junctions at newly formed cell-cell contacts. We found that individual small gap junctions were very dynamic, moving about or becoming assembled and disassembled in the course of minutes. Larger gap junctions were more stable than small punctate ones. In control condition, stable new gap junctions were not formed during observation times of 30 min or longer. But at elevated levels of cyclic adenosine monophosphate, the chimeric channel protein began aggregating at new junctions 5 10 minutes after cell-cell contact and continued to concentrate there for at least one hour. Also already established junctions grew in size. The fluorescent chimeric channel protein will be an excellent tool to investigate the regulation of trafficking of connexin from and to the membrane and the mechanism of connexin channel aggregation into gap junctions. PMID- 10669105 TI - Transforming growth factor-beta1 regulates basement membrane formation by alveolar epithelial cells in vitro. AB - Immortalized alveolar type II epithelial (SV40-T2) cells formed a continuous, thin lamina densa when they were cultured on collagen fibrils with the supplement of 1.0 ng/ml TGF-beta1. Corresponding to lamina densa formation, immunohistochemical analysis of laminin, type IV collagen, perlecan, and entactin (nidogen) indicated integration of these components in a linear array beneath the SV40-T2 cells. Synthesis of these basement membrane constituents was significantly enhanced by TGF-beta1 in a dose-dependent manner. On the other hand, TGF-beta1 did not affect the synthesis of extracellular matrix-regulatory enzymes and their inhibitors, such as type II transglutaminase, matrix metalloproteinase-2, plasminogen activator inhibitor-1, or tissue inhibitor of matrix metalloproteinase-1. These results indicate that basement membrane formation in the presence of 1.0 ng/ml TGF-beta1 is attributable to enhanced synthesis of basement membrane constituents. However, formation of a continuous basement membrane was inhibited at a TGF-beta1 concentration of 5.0 ng/ml. Synthesis of the basement membrane constituents was further enhanced at this concentration and the extracellular matrix-regulatory enzymes remained unchanged. The deposits of cellular fibronectin and type I collagen beneath SV40-T2 cells were significantly augmented. Thus excessive production of interstitial extracellular matrix components appears to obstruct the integration of basement membrane constituents into a continuous architecture. These results indicate that the basement membrane formation by SV40-T2 cells is achieved at the optimal TGF beta1 concentration. PMID- 10669106 TI - Activity-independent cell adhesion to tissue-type transglutaminase is mediated by alpha4beta1 integrin. AB - Transglutaminases (TGases) are enzymes which catalyze cross-link formation between glutamine residues and lysine residues in substrate proteins. We have previously reported that one of the TGases, blood coagulation factor XIIIa (FXIIIa), is capable of mediating adhesion of various cells. In this paper, we report for the first time that tissue-type transglutaminase (TGc) also has cell adhesion activity. TGc-coated plastic surface promoted adhesion and spreading of cells in a TGc concentration-dependent manner. However, there are some obvious differences between cell adhesion mediated by TGc and FXIIIa. As was reported previously, the adhesion to FXIIIa is dependent on its TGase activity. In contrast, the TGc-mediated cell adhesion is independent of its TGase activity: 1) The modification of the active center cysteine with iodoacetamide blocked the enzyme activity without any effect on cell adhesion; 2) the addition of Mg2+ did not induce the enzyme activity, but it was as effective as Ca2+ for cell adhesion; 3) the addition of NH4+ inhibited the enzyme activity but did not affect the cell adhesion significantly. The integrins involved in these cell adhesions are quite different. In the case of FXIIIa, alpha vbeta3 and alpha5beta1 integrins are involved and consequently the RGD peptide substantially inhibited the adhesion. On the other hand, the cell adhesion to TGc is mediated by alpha4beta1 integrin but not alpha5beta1; a CS-1 peptide, which represents the binding site of fibronectin to alpha4beta1 integrin, completely inhibited the cell adhesion to TGc. It is possible that TGc and FXIIIa may mediate cell adhesion under different physiological and pathological situations. PMID- 10669107 TI - Carboxypeptidase E does not mediate von Willebrand factor targeting to storage granules. AB - Sorting of von Willebrand factor precursor (pro-vWf) from the trans-Golgi network to secretory granules (Weibel-Palade bodies) is critical for its conversion to the biologically active highly multimeric form, as well as for regulated secretion by the endothelial cells. When expressed in hormone-secretory cells, vWf is also recognized as a stored protein and is directed to storage granules. Recently, carboxypeptidase E (CPE) was proposed as a granular sorting receptor for prohormones (Cool et al., Cell 88: 73, 1997). To explore whether CPE is also involved in pro-vWf sorting, we initially examined its expression in human umbilical vein endothelial cells. A specific message for CPE and the protein itself were detected making it a plausible candidate as a targeting receptor for vWf in endothelium. To investigate this possibility, we used mice lacking CPE. The highly multimeric forms, subunit composition and plasma levels of vWf in CPE deficient mice were similar to those of their wild-type littermates. vWf was also found in alpha-granules of platelets and in Weibel-Palade bodies of endothelial cells obtained from the CPE-deficient mice. Furthermore, vWf was released from the cultured CPE-deficient endothelial cells after stimulation with a secretagogue. We conclude that CPE is not essential for sorting vWf to the regulated secretory pathway. Thus, a CPE-independent mechanism must exist for protein sorting to storage granules. PMID- 10669108 TI - Comparative analysis of Ca2+ and H+ flux magnitude and location along growing hyphae of Saprolegnia ferax and Neurospora crassa. AB - Calcium and proton ion fluxes were mapped at the growing apices of two hyphal organisms, the oomycete Saprolegnia ferax and the ascomycete Neurospora crassa and pseudohyphal Saccharomyces cerevisiae using self-referencing ion-selective probes. S. ferax exhibited well-defined transport zones absent in N. crassa. Ca2+ fluxes were located within 8 microm of the growing hyphal tip; the net Ca2+ flux was either inward (75% of all experiments) or outward. The inward component of the net flux was inhibited by Gd3+, known to inhibit Ca2+ permeable stretch activated channels. Because the Ca2+ flux is located at the region of maximal hyphal expansion, exocytosis may contribute to Ca2+ efflux, in addition to the stretch-activated channel mediated influx. Maximal inward H+ flux was observed 10 30 microm behind the hyphal tip where peak mitochondria densities taper off at the onset of a vacuolation zone, presumably due to highly localized H+ cotransporter activity. By contrast, N. crassa exhibited no net Ca2+ flux and a consistently inward H+ flux (93% of all experiments) that was homogeneously distributed up to 60 microm behind the hyphal apex. Both hyphal organisms have similar tip morphology and growth rates, and are reported to have tip-high cytosolic Ca2+ gradients associated with growth. Only S. ferax exhibited tip localized Ca2+ fluxes and a well defined H+ influx zone just behind the tip. Differences in ecological habitats and cytology--S. ferax is an aquatic organism that grows as a migrating plug of cytoplasm while N. crassa is normally terrestrial with a cytoplasm-rich mycelium and highly active cytoplasmic streaming behind the growing margin--may account for the differences in the 'architecture' of ion transport occurring during the process of tip growth. Net Ca2+ efflux and H+ influx of growing S. cerevisiae pseudohyphae were also measured but localization was not possible due to small cell size. PMID- 10669109 TI - Caulerpenyne blocks MBP kinase activation controlling mitosis in sea urchin eggs. AB - In a previous study, we demonstrated that caulerpenyne (Cyn), a natural sesquiterpene having an antiproliferative potency, blocked the mitotic cycle of sea urchin embryos at metaphase and inhibited the phosphorylation of several proteins, but did not affect histone H1 kinase activation (Pesando et al, 1998, Eur. J. Cell Biol. 77, 19-26). Here, we show that concentrations of Cyn that blocked the first division of the sea urchin Paracentrotus lividus embryos in a metaphase-like stage (45 microM) also inhibited the stimulation of mitogen activated protein kinase (MAPK) activity in vivo as measured in treated egg extracts using myelin basic protein (MBP) as a substrate (MBPK). However, Cyn had no effect on MBP phosphorylation when added in vitro to an untreated egg extract taken at the time of metaphase, suggesting that Cyn acts on an upstream activation process. PD 98059 (40 microM), a previously characterized specific synthetic inhibitor of MAPK/extracellular signal-regulated kinase-1 (MEK1), also blocked sea urchin eggs at metaphase in a way very similar to Cyn. Both molecules induced similar inhibitory effects on MBP kinase activation in vivo, but had no direct effect on MBP kinase activity in vitro, whereas they did not affect H1 kinase activation neither in vivo nor in vitro. As a comparison, butyrolactone 1 (100 microM), a known inhibitor of H1 kinase activity, did inhibit H1 kinase of sea urchin eggs in vivo and in vitro, and blocked the sea urchin embryo mitotic cycle much before metaphase. Immunoblots of mitotic extracts, treated with anti active MAP-kinase antibody, showed that both Cyn and PD 98059 reduced the phosphorylation of p42 MAP kinase (Erk2) in vivo. Our overall results suggest that Cyn blocks the sea urchin embryo mitotic cycle at metaphase by inhibiting an upstream phosphorylation event in the MBPK activation pathway. They also show that H1 kinase and MBPK activation can be dissociated from each other in this model system. PMID- 10669110 TI - Actin-rich structures formed during the invasion of cultured cells by infective forms of Trypanosoma cruzi. AB - Previous work has shown that Trypanosoma cruzi extracellular amastigotes as well as metacyclic trypomastigotes infect cultured cells in a highly specific parasite form-cell type interaction. In this work we have investigated the mode of interaction of both forms with HeLa and Vero cells using scanning electron and confocal fluorescence microscopy. We examined the distribution of several host cell components as well as extracellular matrix elements during cell invasion by both T. cruzi infective forms. Scanning electron microscopy showed that membrane expansions formed during the invasion of cells by extracellular amastigotes. These expansions correspond to small cup-like structures in HeLa cells and are comparatively larger "crater"-like in Vero cells. We detected by confocal microscopy actin-rich structures associated with the internalisation of both infective forms of the parasite that correspond to the membrane expansions. Confocal fluorescence microscopy combining DIC images of cells labelled with monoclonal antibodies to phosphotyrosine, cytoskeletal elements, integrins, and extracellular matrix components revealed that some of the components like gelsolin and alpha-actinin accumulate in actin-rich structures formed in the invasion of amastigotes of both cell types. Others, like vinculin and alpha2 integrin may be present in these structures without evident accumulation. And finally, some actin-rich processes may be devoid of components like fibronectin or alphaV integrin. These studies provide evidence that the repertoire of host cell/extracellular matrix components that engage in the invasion process of T. cruzi forms is cell type- and parasite form-dependent. PMID- 10669111 TI - The Lps locus: genetic regulation of host responses to bacterial lipopolysaccharide. AB - Lipopolysaccharide (LPS), an abundant glycolipid of the outer membrane of gram negative bacteria, is able to provoke a generalized proinflammatory response in the infected host. Genetic regulation of this trait has been localized to the Lps locus on mouse chromosome 4. Several inbred mouse strains, including C3H/HeJ, C57BL/10ScNCr and C57BL/10ScCr, bear mutations at the Lps locus (Lps(d)) that confer hyporesponsiveness to the immunostimulatory properties of LPS and susceptibility to overwhelming gram-negative bacterial infection. The phenotypic expression of Lps(d) is pleiotropic, affecting several cell types crucial to host defense, including the macrophage. By positional cloning, Toll-like receptor 4 (Tlr4), a transmembrane protein with a cytoplasmic domain that bears homology to the Interleukin-1 receptor, has been identified as the gene encoded by Lps. Tlr4 is a member of a novel gene family that participates in host defense against microbial infection in plants, invertebrates and mammals. Discovery of the molecular basis of the Lps mutation represents a significant advance in defining the fundamental mechanisms of cellular activation by LPS. PMID- 10669112 TI - A new class of COX-2 inhibitor, rutaecarpine from Evodia rutaecarpa. AB - OBJECTIVE AND DESIGN: We investigated the effect of a new class of COX-2 inhibitor, rutaecarpine, on the production of PGD2 in bone marrow derived mast cells (BMMC) and PGE2 in COX-2 transfected HEK293 cells. Inflammation was induced by lambda-carrageenan in male Splague-Dawley (SD) rats. MATERIAL: Rutaecarpine (8,13-Dihydroindolo[2',3':3,4]pyridol[2,1-b]quinazolin -5(7H)-one) was isolated from the fruits of Evodia rutaecarpa. BMMC were cultured with WEHI-3 conditioned medium. c-Kit ligand and IL-10 were obtained by their expression in baculovirus. METHODS: The generation of PGD2 and PGE2 were determined by their assay kit. COX 1 and COX-2 protein and mRNA expression was determined by BMMC in the presence of KL, LPS and IL-10. TREATMENT: Rutaecarpine and indomethacin dissolved in 0.1% carboxymethyl cellulose was administered intraperitoneally and, 1 h later, lambda carrageenan solution was injected to right hind paw of rats. Paw volumes were measured using plethysmometer 5 h after lambda-carrageenan injection. RESULTS: Rutaecarpine inhibited COX-2 and COX-1 dependent phases of PGD2 generation in BMMC in a concentration-dependent manner with an IC50 of 0.28 microM and 8.7 microM, respectively. It inhibited COX-2-dependent conversion of exogenous arachidonic acid to PGE2 in a dose-dependent manner by the COX-2-transfected HEK293 cells. However, rutaecarpine inhibited neither PLA2 and COX-1 activity nor COX-2 protein and mRNA expression up to the concentration of 30 microM in BMMC, indicating that rutaecarpine directly inhibited COX-2 activity. Furthermore, rutaecarpine showed in vivo anti-inflammatory activity on rat lambda-carrageenan induced paw edema by intraperitoneal administration. CONCLUSION: Anti inflammatory activity of Evodia rutaecarpa could be attributed at least in part by inhibition of COS-2. PMID- 10669113 TI - Eosinophils and neutrophils in biopsies from the middle ear of atopic children with otitis media with effusion. AB - OBJECTIVE AND DESIGN: The majority of patients with otitis media with effusion (OME) and atopy have been shown to have elevated levels of eosinophil cationic protein (ECP) in their middle ear fluid. The mechanism underlying these elevated levels of ECP is not clear. The purpose of this study was to investigate the feasibility of a quantitative determination of eosinophils and neutrophils in the middle ear lining by specific immunocytochemical markers, in order to study the extent of the involvement of these cells in patients with OME. METHODS: Bilateral middle ear biopsies from five children with persistent OME and atopy confirmed by in vitro testing were evaluated for the presence of eosinophils and neutrophils with monoclonal antibodies against specific granule proteins. Five subjects who had no signs of effusion or infection but were undergoing routine tympanoplasty for dry perforations served as controls. The biopsies were embedded in a plastic resin to improve the structural preservation of the target cells and to increase the resolution in the light microscope. Dual markers were used to determine which marker was better for eosinophils and neutrophils, respectively. The following markers were used: eosinophil cationic protein (EG2), and eosinophil peroxidase (EPO) for eosinophils and myeloperoxidase (MPO) and human neutrophil lipocalin (HNL) for neutrophils. RESULTS: Antibodies against EPO gave a more localized and intense staining than antibodies against EG2. Antibodies against HNL appear more specific to neutrophils than antibodies against MPO that also recognize monocytes. The number of cells was determined both in the tissue and in the mucus covering the epithelium. Eosinophils and neutrophils were present in the subepithelial connective tissue and in the mucus blanket in the middle ear of patients with OME in significantly higher number than in the control group. In general, there were more inflammatory cells in the mucus than in the tissue itself, but the number of inflammatory cells in the mucus showed a significant positive correlation with the number of inflammatory cells in the tissue. There was a significant positive correlation between the number of neutrophils and the number of eosinophils in the tissue as well as in the mucus, irrespective of which marker was used. CONCLUSION: The results of this study show the feasibility of using specific antibodies to identify eosinophils and neutrophils in the middle ear. The initial data suggest that atopic children with OME have higher numbers of such cells as compared to non-OME controls. PMID- 10669114 TI - Eicosanoid release in the endotoxin-primed isolated perfused rat lung and its pharmacological modification. AB - OBJECTIVE: Recent observations have demonstrated a central role of the "inducible" isoform of the cyclooxygenase (COX), COX-2, in the rat lung. Therefore, the reported capacity of selective COX-2 inhibitors to potentiate the formation of leukotriene (LT) B4 may raise concern about pro-inflammatory side effects of such drugs in the respiratory system. The present study was aimed at determining the effects of the COX-2 inhibitor NS-398 on the release of COX and 5 lipoxygenase (LOX) metabolites of arachidonic acid in isolated perfused lungs obtained from endotoxin-treated rats before and after stimulation with the leukocyte secretagogue N-formyl-methionyl-leucyl-phenylalanine (FMLP). METHODS: Two hours after rats had received endotoxin i.v., the lung was dissected and perfused via the pulmonary artery with physiological salt solution. After an equilibration period of 20 min the outflow was collected (5-min fractions). In the respective treatment groups, indomethacin, NS-398, or the 5-LOX inhibitor MK886 were present throughout the experiment, while FMLP was added to the perfusate during a single 5-min period. The concentration of eicosanoids in the outflow was determined by radioimmunoassay. RESULTS: Endotoxin treatment of rats resulted in increased expression of COX-2 mRNA in lung tissue, and an elevated basal release of the prostaglandin (PG)I2 metabolite 6-keto PGF1alpha, without a detectable increase of leukotriene (LT) formation. In-vitro exposure to FMLP stimulated LT and prostanoid release, which was significantly enhanced in endotoxin-primed lungs, and was suppressed by the 5-LOX inhibitor MK-886 (3 microM) and the COX-inhibitor indomethacin (5 microM), respectively. Either compound showed selective inhibition of the respective pathway of arachidonic acid metabolism. In endotoxin-primed lungs, the COX-2 inhibitor NS-398 (0.3-1.0 microM) depressed basal as well as FMLP-stimulated release of 6-keto PGF1alpha, but did not cause a significant increase of LTB4 or cysteinyl-LT release. CONCLUSIONS: These results suggest that FMLP, presumably acting on inflammatory cells trapped in the pulmonary circulation of endotoxin treated rats, induced prostanoid formation mainly via the COX-2 pathway, and that its inhibition by NS 398 had no detectable potentiating effect on LTB4 or cysteinyl-LT biosynthesis. PMID- 10669115 TI - Chemoattractant-induced release of elastase by tumor necrosis factor-primed human neutrophils: auto-regulation by endogenous adenosine. AB - OBJECTIVE AND DESIGN: In the present work, we studied the role of cell-derived adenosine in both the physiologic regulation and pharmacologic control of the exocytosis of azurophilic granules of neutrophils exposed to tumor necrosis factor alpha (TNF) and stimulated with some chemoattractants. MATERIAL AND METHODS: Human neutrophils were pre-incubated in the absence or presence of TNF. Thereafter, the appropriate chemoattractant was added to the cells. After incubation, the cell-free supernatant was collected for testing elastase activity and intracellular cAMP levels. Results, expressed as mean +/- 1 SD, were evaluated by unpaired, two-tailed Student's t-test and by analysis of variance followed by Student-Newman-Keuls multiple comparisons test. RESULTS: Neutrophil incubation with 10 ng/ml TNF or 0.1 micromol/l N-formyl-met-leu-phe (fMLP) failed to release elastase activity (NE) (NE in absence of stimulus: 23.1 +/- 5.7 nmol/h; TNF-induced NE: 26.4 +/- 14.4 nmol/h; fMLP-induced NE: 27.0 +/- 9.9 nmol/h). Neutrophils, pre-exposed to various amounts of TNF, released elastase in response to 0.1 micromol/l fMLP in a dose-dependent manner (NE in presence of 10 ng/ml TNF and 0.1 micromol/l fMLP: 133.7 +/- 24.0 nmoles/h). As compared with fMLP, C5a had lower activity (NE in presence of 10 ng/ml TNF and 0.1 micromol/l C5a: 66.4 +/- 25.1 nmoles/h), whereas interleukin-8, platelet activating factor and leukotriene B4 were ineffective. The secretory response of TNF-primed neutrophils to fMLP was inhibited by adenosine in a dose-dependent manner (IC50 = 5.18 +/- 7.1 micromol/l). The addition of adenosine deaminase (ADA) to TNF-primed neutrophils resulted in increased secretory response to fMLP (NE in absence and presence of 0.25 U/ml ADA: 71.5 +/- 11.0 and 107.3 +/- 18.6 respectively, P = 0.060). Moreover, two inhibitors of phosphodiesterase type IV (RO 20-1724 and nimesulide) reduced the elastase release only in the absence of ADA (RO 20-1724: percent inhibition in absence or presence of ADA = 20.2 +/- 15.0 and 4.4 +/- 5.1 respectively; nimesulide: percent inhibition in absence or presence of ADA = 22.2 +/- 19.6 and 0.8 +/- 3.0 respectively). Similarly, RO 20-1724 and nimesulide increased intracellular cAMP levels only in absence of ADA (RO 20-1724: percent cAMP increment in absence or presence of ADA = 215.4 +/- 97.5 and 47.3 +/- 53.3 respectively; nimesulide: percent cAMP increment in absence or presence of ADA = 177.7 +/- 19.0 and 19.5 +/- 29.3 respectively). CONCLUSIONS: Endogenous adenosine down-regulates the cell secretory response and is instrumental in uncovering the susceptibility of azurophilic granule exocytosis to control by inhibitors of phosphodiesterase type IV. PMID- 10669116 TI - Differential effects of interleukin-4 and interleukin-10 on nitric oxide production by murine macrophages. AB - OBJECTIVE: To study the effect of interleukin (IL)-4 and IL-10 on nitric oxide (NO) production by macrophages. MATERIALS AND METHODS: Elicited or resident peritoneal macrophages (PMO) and a macrophage cell line Raw264.7 were primed by IL-4 or IL-10 for 6 hours, and were further incubated in the presence of interferon (IFN)-gamma and/or lipopolysaccharide (LPS) for 48 hours. NO2- accumulation in the supernatant of cultured cells was used as an indicator of NO production and was determined by the standard Griess reaction adapted for microplates. The amount of tumor necrosis factor (TNF)-alpha in the culture supernatants was determined with a commercially available ELISA kit. The absorbance was measured at 450 nm with a microplate photometer. RESULTS: IL-4 inhibited NO production by murine macrophages of different sources and the macrophage cell line Raw264.7. In contrast, different macrophage populations showed differential responses to IL-10. After stimulation with LPS or IFN-gamma, IL-10 suppressed NO production by elicited PMO but enhanced NO production by resident PMO or by Raw264.7. Both IL-4 and IL-10 inhibited the production of TNF alpha, which has been shown to play a crucial role in NO production. In the presence or the absence of blocking antibody to TNF-alpha, IL-10 always enhanced NO production by resident PMO. This result suggests that the inhibition of TNF alpha production and the enhancement of NO production by resident PMO stimulated with IL-10 are independent, coexisting events. CONCLUSIONS: Factors other than TNF-alpha have been suspected to influence NO production by macrophages, and this study indicates that IL-10 may be a candidate cytokine for resident PMO. PMID- 10669117 TI - Detection of an approximately 100 kD protein with strong immunoreactivity to antibodies specific for inducible nitric oxide synthase (iNOS) but without NOS activity in neutrophils of patients suffering from sepsis: results of a preliminary study. AB - OBJECTIVE: The study was designed to evaluate the expression of inducible nitric oxide synthase (iNOS) in activated human neutrophils. SUBJECTS AND METHODS: By Western blotting and immunocytochemical staining, iNOS expression was analyzed in neutrophils from patients with sepsis who we classified by high plasma nitrate as subjects with activated NO metabolism. For comparison, rats treated with Zymosan documented to induce iNOS were analyzed. RESULTS: Strong immunoreactivity to antibodies for iNOS was detected in leukocytes of Zymosan treated rats and in neutrophils of septic patients. Such immunoreactivity was absent in untreated rats and healthy subjects. In rats, the immunoreactivity was associated to the 130 kD protein as expected for iNOS. In contrast, the intense iNOS staining in neutrophils of septic patients was associated to an approximately 100 kD protein. Despite the iNOS staining, no increased NOS activity could be demonstrated in the neutrophils of septic patients. CONCLUSIONS: The detection of an approximately 100 kD protein with immunoreactivity to antibodies recognizing human iNOS but without NOS activity in neutrophils of patients with sepsis should initiate studies to elucidate the origin and function of this protein. PMID- 10669118 TI - Glucocorticoids, antioxidants and staurosporine modulate the adherence between monocytes and malaria infected erythrocytes. AB - OBJECTIVE AND DESIGN: Adherence interactions involving monocytes are important for cell-cell interaction as an integral component of immune function. The adherence of malaria parasitised red cells to monocytes was determined after monocytes were treated with dexamethasone, cortisol, ambroxol, danazol, probucol and staurosporine. MATERIALS: Human peripheral blood monocytes isolated by density gradient centrifugation and adherence to glass cover slips. METHODS: The adherence of malaria parasitised red cells to monocytes was determined after the monocytes were incubated for 24 h in the presence of each of the 6 drugs. RESULTS: The two glucocorticoids and staurosporine reduced the adherence of malaria infected erythrocytes to monocytes in a dose dependent manner at concentrations from 10(-10) M and above and ambroxol, danazol, and probucol at 10(-5) M. Staurosporine was the most effective of the drugs studied, completely abolishing adherence at 10(-6) M. CONCLUSION: The adherence of malaria infected erythrocytes to monocytes is reduced in response to glucocorticoids (dexamethasone and cortisol), anti-oxidants (probucol and ambroxol), danazol and staurosporine. PMID- 10669119 TI - Pharmacological modulation of the IFNgamma-induced accessory function of alveolar macrophages and peripheral blood monocytes. AB - OBJECTIVE AND DESIGN: Although alveolar macrophages (AM) are poor accessory cells, alveolar macrophages of patients with sarcoidosis or tuberculosis show an elevated accessory function indicating that the accessory function (AF) of AM can be upregulated. MATERIALS AND METHODS: We examined whether the AF of AM and peripheral blood monocytes (PBM) can be increased by interferon-gamma (IFNgamma) and whether immunomodulating drugs like cyclosporine A, cyclophosphamide, dexamethasone, and ambroxol are able to modulate the accessory function and the expression of accessory molecules. RESULTS: IFNgamma increased the AF of AM and PBM up to 309 +/- 122% and 152 +/- 25%, respectively (p < 0.02, unstimulated control = 100%, in all cases). In alveolar macrophages this increase is most efficiently prevented by cyclosporine A (31 +/- 13%), followed by cyclophosphamide (64 +/- 20%) and dexamethasone (66 +/- 20%). In monocytes the IFNgamma-induced increase in accessory function is prevented only by cyclosporine A (17 +/- 7%) and dexamethasone (59 +/- 9%). Cyclosporine A was also the most effective drug downregulating the expression of accessory molecules (CD54, CD58, CD80 and CD86). CONCLUSIONS: In summary, the accessory function of alveolar macrophages and monocytes is upregulated by IFNgamma and can be controled by immunomodulating drugs. PMID- 10669120 TI - Trends in presentation of congenital heart disease in a population-based study in Malta. AB - Differing pathological haemodynamics in cardiac malformations lead to varying modes and timings of presentation. This study identifies historical trends in presentation of congenital heart disease in a population-based study. All patients diagnosed as having congenital heart disease in Malta between 1960-1994 were included (n = 868). Analysis was carried out on trends in referral sources, modes of presentation and birth prevalence. The number of patients diagnosed with congenital heart disease increased over the period under study. For both patients not requiring intervention (n = 283) and those requiring intervention (n = 585), the proportion diagnosed prior to hospital discharge increased (p < or = 0.005). There was a decreasing trend for general practitioners to refer cases (p < 0.0001), and an increasing trend for paediatricians to refer such patients (p < or = 0.0003). The commonest presentation to the general practitioner was an incidental finding (92%), while paediatricians referred more patients for cyanosis or heart failure (p < or =, 0.005). For lesions not requiring intervention, the commonest lesion referred was ventricular septal defect from all sources. For lesions requiring intervention, the commonest lesion detected prior to hospital discharge was tetralogy of Fallot. Atrial septal defects were the commonest lesions detected after discharge by both paediatricians and general practitioners. An increase in the proportion of hospital diagnoses is attributed to increasing rate of hospital delivery, and greater training and experience in doctors performing neonatal examinations prior to discharge. Patients diagnosed after discharge are increasingly diagnosed by paediatricians due to an increasing pool of paediatricians and better parent awareness and education. PMID- 10669121 TI - Time trends in incidence of cervical cancer in Lithuania from 1983 to 1997. AB - This report examines time trends in both age-specific and age-standardized cervical cancer (ICD9-180) incidence rates among Lithuanian women. The study covers the period from 1983 to 1997. In case of the age-specific rates Poisson regression revealed the controversial results for the different age groups. The incidence rates increased (beta > 0) for the women below 50 years and decreased (beta < 0) for the women above 50 years. The average annual percentage changes were estimated using the age-drift model. In the age groups from 30 to 49 years old the incidence rate increased by 3.3% per year with the 95% confidence interval (CI) being 2.2-4.4. In the age groups from 50 to 69 years old the incidence rate decreased by 2.9% (95% CI: 1.9-3.8). In case of the age standardized rates during the last few years some increase was observed, although no strict conclusions could be drawn. This observation together with the results of the age-specific rate studies predicts that in the near future the age standardized rate will start growing. PMID- 10669122 TI - Antihypertensive drug therapy and hypoglycemia in elderly diabetic patients treated with insulin and/or sulfonylureas. Gruppo Italiano di Farmacovigilanza nell'Anziano (GIFA). AB - We performed this case control study to evaluate the risk of hypoglycemia associated with the use of antihypertensive drugs in older hospitalized diabetic patients treated with sulfonylureas and/or insulin. All diabetic patients admitted during 4 months in 1988, month in 1991, 4 months in 1993 and 4 months in 1995 (n = 3477, mean age 71.4 +/- 0.2 years, 1542 males and 1935 females) were enrolled in the study. During the four annual surveys 86 patients (mean age 71.1 +/- 1.4 years, 33 males and 53 females) presented hypoglycemia during hospital stay. The patients who presented hypoglycemia were less frequently users of sulfonylureas and more frequently users of a combination of insulin and sulfonylureas. Use of antihypertensive drugs was similar in the two groups studied, and among potentially interacting drugs considered in the analysis, sulfonamides were more frequently used in patients who experienced hypoglycemia. Moreover, patients with hypoglycemia used a higher number of drugs, had a longer length of stay and had a greater prevalence of hypoglycemia as admission problem. Finally, although not significant, liver and renal diseases were more frequent among patients with hypoglycemia. In the multivariate analysis, contemporary use of insulin and sulfonylureas, liver disease and length of stay were significantly associated with hypoglycemia, while none of the antihypertensive drugs showed a significant association with the occurrence of hypoglycemia during hospital stay. Our results indicate that antihypertensive drugs do not increase the risk of hypoglycemia in elderly diabetic patients. PMID- 10669123 TI - Prevalence of antibodies to hepatitis A among children and adolescents in Larnaca area, Cyprus. AB - The prevalence of antibodies to hepatitis A virus was investigated in 385 children and adolescents (52.2% males), aged 6 to 18, in the Larnaca area of Cyprus. This is the first study investigating the prevalence of hepatitis A in Cyprus for this age group. The population was stratified into two groups: 6 to 12 years old and 13 to 18 years old. None of the subjects in the first group were positive. The prevalence of hepatitis A in the age of group 13 tol8 was 1.6%. In conclusion, the low prevalence of anti-HAV demonstrates the susceptibility of young Cypriots to hepatitis A. This is a cause for concern as these unprotected young adults are frequently exposed to potentially infected individuals. PMID- 10669124 TI - Maternal knowledge and environmental factors associated with risk of diarrhea in Israeli Bedouin children. AB - Diarrhea is still a major cause of morbidity and mortality among children in developing countries. As it is due to multiple causative agents including viruses, bacteria and parasites, biological interventions are not currently available to markedly reduce incidence and severity. We examined maternal knowledge and reported behavior during diarrheal episodes, as well as environmental factors to determine their association with diarrhea. The children and mothers were from a Bedouin township in southern Israel, which has developed preventive and curative health care facilities. The Bedouin population in Israel is in transition from a nomadic to a settled life style. While almost all mothers exhibited good knowledge regarding food storage and prevention of diarrhea episodes in the children, the rate of illness in the children remained relatively high (two episodes per child year of observation). In a multivariate analysis, cessation of breastfeeding during diarrhea, child sleeping with siblings and lack knowledge about risk factors, were the major risk factors for illness with odds ratios (OR): 4.6, p = 0.02, 5.6, p = 0.03 and 1.7, p = 0.06, respectively. These data indicate that even in this population with free access to preventive medical care, greater efforts should be made to educate mothers regarding risk factor for diarrheal disease identification and the benefits of maintaining breastfeeding during diarrhea episodes. PMID- 10669125 TI - Prevalence of bacterial vaginosis and correlation of clinical to Gram stain diagnostic criteria in low risk pregnant women. AB - The present study aimed to evaluate the prevalence of bacterial vaginosis, and the correlation of clinical Amsel criteria with Gram Nugent criteria for the diagnosis of bacterial vaginosis in a low risk population of pregnant women. Pregnant women under 28 weeks of gestation who were followed in the low risk clinics at two centers were evaluated for the presence of bacterial vaginosis using the Amsel clinical criteria, and underwent vaginal samples for Gram staining. Gram smears were examined for the diagnosis of bacterial vaginosis on the basis of Nugent's criteria. A total of 492 women were included in the study. Bacterial vaginosis was diagnosed in 1.6% (8/492) women on the basis of clinical criteria, and in 4.5% (22/492) according to Gram stain. The sensitivity and specificity of Amsel criteria compared with Gram stain were 35% and 99%, respectively. In accordance with other recent reports, the prevalence of bacterial vaginosis appears to be much lower in certain areas than figures previously suggested. In these populations, the correlation of composite clinical criteria defined in groups with high prevalence of bacterial vaginosis appears to be also poor. PMID- 10669126 TI - Botulism surveillance in Italy: 1992-1996. AB - Though a relatively rare disease, botulism can be a serious problem of public health, particularly when connected with the consumption of industrial canned food; moreover, in the last years the shortage of botulism antitoxin has caused some concern in the Public Health Authorities. This work presents the results of a five-year surveillance of botulism in Italy, with the distribution of the cases by Regions (first level administrative units which, in Italy, have administrative and legislative competencies in the sanitary field) and by vehicle of transmission. All the relevant and confirmed botulism outbreaks that occurred in the period under consideration are described. PMID- 10669127 TI - A review of zygomycosis due to Basidiobolus ranarum. AB - Zygomycosis due to Basidiobolus ranarum (entomophthoromycosis basidiobolae, subcutaneous zygomycosis, subcutaneous phycomycosis, basidiobolomycosis) is a granulomatous infection of the skin and subcutaneous tissues characterized by the formation of fluctuant firm and non-tender swellings, generally on the extremities, trunk and rarely other parts of the body. The causative agent is common in soil, decaying vegetable matter, and the gastrointestinal tracts of amphibians, reptiles, fish and bats. It is presumed that infection is acquired through exposure to B. ranarum following minor trauma to skin or insect bites. The disease usually occurs in children, less often in adolescents and rarely in adults. Males are much more frequently affected than females. Laboratory diagnosis is based on histopathology and culture. The typical histopathological feature is the presence of thin-walled, broad often aseptate hyphae or hyphal fragments with an eosinophilic sheath, frequently phagocytized within giant cells. Basidiobolus ranarum is known to produce several enzymes, e.g. lipase and protease that probably play roles in the pathogenesis of infections caused by this mould. An immunological test has been developed for specific diagnosis of the disease. Though potassium iodide (KI) has been the traditional drug employed in the treatment of infections by B. ranarum, several other drugs, viz amphotericin B, cotrimoxazole, ketoconazole, itraconazole and fluconazole have been successfully tried. PMID- 10669128 TI - Canine neosporosis: clinical and pathological findings and first isolation of Neospora caninum in Germany. AB - Neosporosis was diagnosed in an 11-week-old puppy of the breed Kleiner Munsterlander with progressive hindlimb paresis. Pathohistological and immunohistological examinations revealed a disseminated infection with Neospora caninum. Parasitic stages were demonstrated in the brain, spinal cord, retina, muscles, thymus, heart, liver, kidney, stomach, adrenal gland, and skin. Immunohistochemistry investigations were carried out using polyclonal rabbit antisera developed against N. caninum tachyzoites and the recombinant bradyzoite specific antigen BAG-5 of Toxoplasma gondii, which is known to cross-react with N. caninum bradyzoites. BAG-5 antibodies recognized tissue cysts within the CNS and some protozoan stages that were not surrounded by a visible cyst wall. All parasite clusters in the retina and some in muscle tissue stained positively with the BAG-5 antiserum. N. caninum was isolated in cell culture and mice inoculated with brain and spinal cord of the puppy. The new isolate is the first reported in Germany and is designated NC-GER1. PMID- 10669129 TI - Blastocystis hominis: origin and significance of vacuolar and granular forms. AB - Using supravital neutral red staining and light microscopy, individual Blastocystis organisms, subcultured from clinical isolates in modified monophasic Robinson's medium, were followed over various periods on glass slides. A rapid transition from uniformly stained to granular and vacuolated forms preceded the organism's death as evidenced by pale staining and Brownian motion in the cell's interior. Granular and vacuolar forms of Blastocystis may indicate degenerative changes in individual cells, fixation artifact, or both. PMID- 10669130 TI - Labeled probes inserted in the macrophage membrane are transferred to the parasite surface and internalized during cell invasion by Toxoplasma gondii. AB - Tachyzoites of Toxoplasma gondii attach to the macrophage surface and are internalized either by a phagocytic process, which can be inhibited by cytochalasin D, or by an active process, independent of host cell actin. Previous studies have shown that parasite attachment induces the secretion of macromolecules found in the apical organelles (micronemes and rhoptries) and subsequent/concomitant parasite internalization with the formation of a membrane bound vacuole known as the parasitophorous vacuole. In the present study we labeled the macrophage surface with fluorescent probes that bind to proteins (DiIC16) and lipids (DTAF) and then allowed control or cytochalasin-D-treated cells to interact with untreated or antibody-coated tachyzoites of T. gondii. The interaction was interrupted at different time points by fixation and the distribution of the probes was analyzed by confocal laser scanning microscopy. Following attachment of the parasites to the macrophage surface, intense labeling of the parasite surface was observed, suggesting transfer of components of the macrophage surface to the parasite surface. Nonadherent parasites were not labeled. Immediately after attachment, most of the parasites were internalized and labeling of the internalized parasites as well as of the parasitophorous vacuole, probably of its membrane, was evident, indicating that surface components of the macrophage are involved in the formation of the parasitophorous vacuole. PMID- 10669131 TI - Ribosomal RNA of Nosema algerae and phylogenetic relationship to other microsporidia. AB - Microsporidia are intracellular parasites that are common in invertebrates. Taxonomic classification is mostly restricted to morphologic and physiologic data. Limited data are available about taxonomic classification using DNA sequence data for analysis. We examined the small-subunit (SSU) rDNA, the intergenic spacer (ITS) region, and a part of the large-subunit (LSU) rDNA of Nosema algerae, a parasite of mosquitoes, taken from a laboratory colony of Anopheles stephensi. Target gene amplifications were done by polymerase chain reaction (PCR) and, after cloning, DNA fragments were sequenced. The SSU-rDNA sequence obtained was aligned with several other microsporidian SSU-rDNA sequences available from the GenBank or EMBL data bases and was analyzed by different methods. On the basis of the results of our phylogenetic analysis, we suggest that our N. algerae isolate is not closely related to other microsporidia belonging to the genus Nosema. PMID- 10669132 TI - A cDNA encoding a nuclear hormone receptor of the steroid/thyroid hormone receptor superfamily from the human parasitic nematode Strongyloides stercoralis. AB - A cDNA encoding a nuclear hormone receptor of the steroid/thyroid receptor superfamily was obtained from third-stage larvae(L3) of the parasitic roundworm Strongyloides stercoralis. A recombinant clone was isolated via screening of an S. stercoralis cDNA library with a polymerase chain reaction (PCR)-generated probe. The insert of 2,583 bp contained the complete coding sequence of the receptor homologue. The conceptually translated amino acid sequence of this open reading frame encodes a 753-amino-acid-residue protein with an apparent molecular weight of 83.6 kDa and a predicted isoelectric point (pI) of 8.52. The coding sequence is 69% AT and the noncoding sequence is 72% AT, reflecting a characteristic A/T codon bias of S. stercoralis. In this report the amino acid sequence of the S. stercoralis nuclear hormone receptor of the steroid/thyroid receptor superfamily is compared with that of nuclear hormones of Caenorhabditis elegans, human orphan nuclear receptors, and insect ecdysone receptors. The potential role of steroids in the induction of hyperinfection syndrome is also discussed. PMID- 10669133 TI - Immunolocalization of two hydrogenosomal enzymes of Trichomonas vaginalis. AB - Three monoclonal antibodies specific for malic enzyme and for the alpha- and beta subunits, respectively, of the succinyl-coenzyme A (CoA) synthetase of Trichomonas vaginalis were used to immunolocalize these proteins in the cell. All antibodies labeled the hydrogenosome matrix as determined both by immunofluorescence and by immunogold staining. There was no labeling on the cell surface or in any other cell compartment. These results support the idea that these proteins are restricted to a hydrogenosomal function and do not play a role as adhesins at the plasma membrane surface. PMID- 10669134 TI - Differential protein kinase C phosphorylation sites in the L17 ribosomal protein from Leishmania infantum. AB - Leishmania infantum, the protozoan parasite responsible for leishmaniasis in Europe, is capable of undergoing developmental changes in vitro and provides an excellent model for the study of cell differentiation processes. We have cloned the gene encoding the L17 ribosomal protein. The LiL17 protein family belongs to the macrolide binding site, related to the peptidyl transferase center of the ribosome. Its comparison with other members of the protein family shows several structural differences that may reflect functional variations. The protein kinase C phosphorylation sites display an intermediate pattern involving differences in location and type of residue with respect to all the species considered. Gene structural analysis suggests the existence of two different encoding genes. The expression of the genes seem to be different with the distinct growth phases of the parasite. PMID- 10669135 TI - Otobothrium cysticum (Cestoda: Trypanorhyncha) from the muscle of butterfishes (Stromateidae). AB - On the basis of the tentacular armature, surface ultrastructure, and morphological measurements of plerocerci obtained from the musculature of butterfishes (Stromateidae), we corroborate an earlier proposal that Otobothrium crenacolle, a commonly reported trypanorhynch cestode from the northwestern Atlantic coast, is a junior synonym of O. cysticum. This action exemplifies at least an Atlantic Ocean and Indian Ocean distribution for O. cysticum. The infection in commercially important butterfishes shows that an otobothriid trypanorhynch may heavily infect fish flesh and influence the market value of some fish species yet also be restricted to the body cavity of other fish intermediate hosts. Infections of O. cysticum in the flesh of Peprilus burti (Gulf butterfish) and P. alepidotus (harvestfish) in the Gulf of Mexico has varied annually since 1970, with samples ranging in prevalence between 20% and 100% and in mean intensity between 1 and 3,500 or more plerocerci per fish. Comparative infections in P. burti from the Gulf of Mexico and P. triacanthus (butterfish) from the Atlantic Ocean demonstrate a present geographic difference in infections. The prevalence and mean intensity in 4 collections of butterfishes ranged from 9% to 98% of the fish and from 1 to 678 plerocerci in a subsample of tissue, respectively, with prevalent and heavy infections being observed in the Gulf of Mexico fish and relatively few individuals being infected with few worms in the Atlantic fish. A slight host response in the butterfishes involving some fatty infiltration and inflammatory infiltration was associated with the metacestode. In some larger fish, encapsulations were yellow, and in a few cases, worms had degenerated. This finding and an increase in intensity with fish weight suggest a continual accumulation of the worms in association with little host resistance. PMID- 10669136 TI - Ultrastructure of the Endolimax nana cyst. AB - This is the first report on the ultrastructure of the Endolimax nana cyst. These cysts are mostly ovoid in shape and have a distinct cyst wall measuring 80 nm. The nuclear membrane is without pores or associated chromatin deposits. The cytoplasm does not have mitochondria or a Golgi apparatus but shows elongated tubular structures made up of a double row of ribosome-like particles. The nature and function of this structure is not known, but it appears to be characteristic of this species and has not been reported from any other intestinal ameba. PMID- 10669138 TI - Restriction-fragment-length polymorphism analysis of small-subunit rRNA genes of Blastocystis isolates from animal hosts. AB - The anaerobic enteric protozoan organism Blastocystis sp. has been identified from mammalian, avian, reptilian, and arthropod hosts. Eight Blastocystis isolates from five animal host species (cow, goat, sheep, guinea pig, and rhea) were compared by small-subunit ribosomal RNA (ssu rRNA) restriction-fragment length polymorphism (RFLP) analyses using five restriction endonucleases. The isolates sorted into five genotypes. Multiple genotypes were found in isolates from a single animal host species, and multiple host species shared a single genotype. A molecular method such as RFLP analysis of ssu rRNA genes facilitates the characterization of Blastocystis isolates from various host species. PMID- 10669137 TI - Restriction-fragment-length polymorphism analysis of small-subunit rRNA genes of Blastocystis hominis isolates from geographically diverse human hosts. AB - Genomic diversity among 14 isolates of Blastocystis hominis from 4 different geographic locations was examined by small-subunit rRNA (ssu rRNA) restriction fragment-length polymorphisms (RFLP) using 5 different restriction endonucleases. On the basis of the observed RFLP patterns among the isolates, a total of 12 genotypes were identified, with 7 isolates exhibiting mixed RFLP genotypes. There was no correlation between B. hominis geographic origin and RFLP banding pattern or genotype. PMID- 10669139 TI - Assessment of a recombinant antigen versus natural hypodermin C for the serodiagnosis of hypodermosis in cattle. AB - An indirect ELISA test using as antigen a recombinant parasite protein, hypodermin C, was developed to measure Hypoderma-specific antibodies in cattle sera and compared with natural hypodermin C. To evaluate the field efficacy of the ELISA test, 334 serum samples were collected from cows raised at farms in Galicia for a serological survey. Compared with an ELISA based on natural parasite antigen, the recombinant hypodermin C gave excellent results, with a sensitivity of 95.8% and a specificity of 95.7%. Considering the cut-off point, with the recombinant hypodermin C, 70.9% of the animals had positive levels of antibodies to Hypoderma and with natural hypodermin C, 73.6%. Recombinant hypodermin C appears to be a useful alternative to the natural parasite antigen for the serodiagnosis of Hypoderma sp in cattle. PMID- 10669140 TI - Variability of Fasciola hepatica infection in Lymnaea ovata in relation to snail population and snail age. AB - Bimiracidial infections of Lymnaea ovata with Fasciola hepatica were performed under laboratory conditions to determine the susceptibility of snails from six French populations to trematode infection. In five populations of L. ovata the prevalence of infection in the 1-mm groups ranged between 2.7% and 43.7% at day 35 postexposure; it decreased in the 2-mm snails and was zero in larger groups. In the snails from Thenay (periodically polluted brook) the prevalence of F. hepatica infection decreased from the 1-mm group to the 8-mm group (from 23.9% to 1.0%) and was zero in the 10-mm L. ovata. The total number of cercariae shed per snail was 18.3 in the 1-mm group, increasing to 117 in the 8-mm group. The latter findings could be interpreted as a consequence of periodic pollution in the brook of Thenay; pollution might disrupt the defense system of L. ovata and facilitate the subsequent larval development of F. hepatica. PMID- 10669141 TI - A monoclonal antibody against Schistosoma haematobium soluble egg antigen: efficacy for diagnosis and monitoring of cure of S. haematobium infection. AB - A monoclonal antibody (mAb), 2F/11F, raised against Schistosoma haematobium soluble egg antigen (SEA) was found to be nonreactive with S. mansoni SEA or other parasite antigens (Fasciola hepatica, Echinococcus granulosus). This IgG1 mAb recognized a repetitive epitope on S. haematobium SEA in the molecular-weight regions of 70, 42, and 35 kDa. It was employed as both an antigen-capture and a biotinylated detection antibody in a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of circulating schistosome antigen (CSA) and had a detection limit of <1 ng S. haematobium SEA/ml. CSA levels were measured in serum and urine samples from 116 S. haematobium-infected rural students before therapy and at 4, 8, and 12 weeks after praziquantel treatment. Serum and urine samples from 50 S. mansoni -infected patients, 15 patients harboring other parasites, and 30 noninfected individuals were also assessed. CSA was detected in 90.5% of serum samples and 94% of urine samples from S. haematobium-infected patients. CSA was undetectable in serum from the 15 patients harboring other parasites and in 94% of serum samples and 84% of urine samples from S. mansoni-infected patients. In the S. haematobium-infected group a positive correlation was detected between CSA levels in serum and urine samples and the egg load per 10 ml urine. A significant reduction in CSA levels was detected in serum and urine samples after praziquantel therapy. CSA was undetectable in 87% of serum samples and 81.5% of urine samples from schistosomiasis haematobium patients at 12 weeks post treatment. These data demonstrate that the use of mAb 2F/11F for detection of CSA provides a sensitive method for the immunodiagnosis and monitoring of cure of schistosomiasis haematobium. PMID- 10669142 TI - Appearance of a stage-specific immunodominant glycoprotein in encysting Entamoeba invadens. AB - The appearance of cyst-specific proteins in encysting Entamoeba invadens and their immunogenicity were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting using an axenic encystation system in vitro. A rabbit antiserum against trophozoites of E. invadens reacted with a number of proteins of cysts after 1-4 days of incubation. Thus, a number of cyst proteins remained antigenically unchanged as common antigens of the two forms after transformation from trophozoites to cysts. A rabbit antiserum against cysts also reacted with the trophozoite proteins as well as the cyst proteins. The most interesting result was that the rabbit anticyst serum reacted predominantly with an 88-kDa protein of cysts after 1 day of incubation. The 88-kDa protein reacted with the anticyst serum absorbed with trophozoite proteins and was thus cyst specific. The reactivity of the 88-kDa protein of cysts with the absorbed anticyst serum decreased as encystation proceeded. When soluble and particulate fractions prepared from cysts after 1 day of incubation were examined by electrophoresis and immunoblotting, the 88-kDa protein that had reacted with the absorbed anticyst serum was found to be present in the particulate fraction, which was rich in cell-wall fragments, and stained with periodic acid-Schiff's reagent, indicating that it is a glycoprotein. The results indicate that encystation is accompanied by appearance of the cyst-specific 88-kDa glycoprotein, which is immunodominant and most abundantly expressed in cysts after 1 day of incubation and appears to be associated with the cyst wall. PMID- 10669143 TI - High levels of factor VIII and venous thrombosis. PMID- 10669144 TI - How much factor V is enough? PMID- 10669145 TI - High plasma concentration of factor VIIIc is a major risk factor for venous thromboembolism. AB - BACKGROUND: Established risk factors, including deficiencies of protein C, protein S or antithrombin and the factor V Leiden and prothrombin mutation, are present in about one third of unselected patients with venous thromboembolism. In addition to these inherited thrombophilic defects, elevated plasma levels of factor VIIIc have been suggested to be important in the pathogenesis of (recurrent) venous thromboembolism. The objective of this study was to assess the relevance of factor VIIIc plasma concentration in consecutive patients with venous thromboembolism. METHOD: We studied the prevalence of elevated plasma levels of factor VIIIc in 65 patients with a proven single episode and in 60 matched patients with documented recurrent venous thromboembolism. The reference group consisted of 60 age- and sex-matched patients who were referred for suspected venous thromboembolism, which was refuted by objective testing and long term clinical follow-up. To minimalize the influence of the acute phase, blood was obtained at least 6 months after the thromboembolic event and results were adjusted for fibrinogen and C-reactive protein. Factor VIIIc was re-determined several years after the first measurement in a subset of patients to evaluate the variability over time. To study a possible genetic cause, a family study was done. FINDINGS: In the control, single and recurrent episode group, the prevalences of plasma levels of factor VIIIc above 175 IU/dl (90th percentile of controls) were 10% (95% CI: 4 to 21%), 19% (95% CI: 10 to 30%) and 33% (95% CI: 22 to 47%), respectively. For each 10 IU/dl increment of factor VIIIc, the risk for a single and recurrent episode of venous thrombosis increased by 10% (95% CI: 0.9 to 21%) and 24% (95% CI: 11 to 38%), respectively. Both low and high plasma levels of factor VIIIc were consistent over time (R = 0.80, p = 0.01). A family study indicated a high concordance for elevated factor VIIIc plasma concentrations among first degree family members. Adjustment for fibrinogen, C reactive protein and known thrombophilic risk factors did not change the observed association of elevated factor VIIIc with thrombosis. INTERPRETATION: Elevated plasma levels of factor VIIIc are a significant, prevalent, independent and dose dependent risk factor for venous thromboembolism. It also predisposes to recurrent venous thromboembolism. PMID- 10669146 TI - Elevation of FVIII: C in venous thromboembolism is persistent and independent of the acute phase response. AB - Recent literature has suggested a role for elevated FVIII:C in venous thromboembolic disease (VTED). However since FVIII:C is known to rise in response to an acute phase reaction, it is difficult to determine whether the increased FVIII:C precedes the thrombosis or represents a secondary reactive phenomenon. In an attempt to address this question, we followed 35 patients with confirmed VTED, raised FVIII:C level (>1.5 iu/ml) and no other thrombotic tendency. Serial measurements of FVIII:C, vWF:Ag, C-reactive protein and fibrinogen were performed. We hypothesized that a persistent increase in FVIII:C in the absence of any other measures of ongoing acute phase response, would support the idea that elevation of FVIII:C is a constitutional phenomenon. Of this initial group, 94% continued to have an elevated FVIII:C level throughout the period of follow up (median 8 months; range 3 to 39 months), with no significant difference between the FVIII:C levels determined at first estimation and those obtained during follow up (p = 0.58). Conversely, only 18% had evidence of an acute phase reaction when first assessed, and nonparametric ranking analysis demonstrated no correlation between FVIII:C and either C-reactive protein or fibrinogen (p = 0.315 and 0.425 respectively).We conclude that increased FVIII:C levels following VTED are persistent, independent of the acute phase reaction, and thus may represent a constitutional risk factor for VTED. PMID- 10669147 TI - Prevention of venous thromboembolism in internal medicine with unfractionated or low-molecular-weight heparins: a meta-analysis of randomised clinical trials. AB - BACKGROUND: The prevention of venous thromboembolic disease is less studied in medical patients than in surgery. METHODS: We performed a meta-analysis of randomised trials studying prophylactic unfractionated heparin (UFH) or low molecular-weight heparin (LMWH) in internal medicine, excluding acute myocardial infarction or ischaemic stroke. Deep-vein thrombosis (DVT) systematically detected at the end of the treatment period, clinical pulmonary embolism (PE), death and major bleeding were recorded. RESULTS: Seven trials comparing a prophylactic heparin treatment to a control (15,095 patients) were selected. A significant decrease in DVT and in clinical PE were observed with heparins as compared to control (risk reductions = 56% and 58% respectively, p <0.001 in both cases), without significant difference in the incidence of major bleedings or deaths. Nine trials comparing LMWH to UFH (4,669 patients) were also included. No significant effect was observed on either DVT, clinical PE or mortality. However LMWH reduced by 52% the risk of major haemorrhage (p = 0.049). CONCLUSIONS: This meta-analysis, based on the pooling of data available for several heparins, shows that heparins are beneficial in the prevention of venous thromboembolism in internal medicine. PMID- 10669148 TI - Factor II G20210A and factor V G1691A gene mutations and peripheral arterial occlusive disease. AB - BACKGROUND: G to A mutations at positions 20210 of the prothrombin gene (F2) and 1691 of the factor V gene (F5) are established risk factors for venous thrombosis. Several factors associated with coagulation and/or fibrinolysis have been associated with arterial occlusive disease, but the role of F2 20210A and F5 1691A for arterial occlusive disease remains unclear. OBJECTIVE: To investigate if F2 20210A and F5 1691A are associated with peripheral arterial occlusive disease (PAOD). METHODS AND RESULTS: We analyzed the prevalence of F2 20210A and F5 1691A alleles in 336 patients with documented PAOD at Fontaine stage II-IV and 300 controls without vascular disease. Allele frequencies in patients and controls were 0.013 and 0.022 for F2 20210A, and 0.042 and 0.045 for F5 1691, respectively, both differences being not statistically significant. CONCLUSION: Our data suggest that mutations F2 G20210A and F5 G1691A are not associated with PAOD. PMID- 10669149 TI - A common polymorphism flanking the ATG initiator codon of GPIb alpha does not affect expression and is not a major risk factor for arterial thrombosis. AB - The platelet membrane glycoprotein (GP) Ib alpha plays a key role in the initial formation of thrombi. Polymorphisms (VNTR and HPA-2) in this receptor are associated with increased risk of coronary heart disease (CHD) and cerebral vascular disease (CVD). We investigated whether a recently described polymorphism (S/R), due to a single base change (T-->C) five nucleotides upstream the initiator codon of GPIb alpha, might influence the expression of the protein, and be implicated in the development of arterial thrombosis. One hundred and thirty nine healthy individuals provided blood samples for DNA analysis of platelet GPIb alpha polymorphisms, and for flow cytometric analysis of the surface expression of the receptor. A group of 20 S/R normal individuals and an identical number of S/S participants, age and sex matched, was investigated for the analysis of the density of various platelet receptors. The distribution of the S/R polymorphism was also analyzed in two case/control studies including 104 CVD patients, 101 CHD patients, and one control age, sex, and environmental risk factors matched for each case patient. Surface density of GPIb alpha showed no wide variations between individuals, was not influenced by the presence of S or R alleles, nor associated with the VNTR or HPA-2 polymorphisms. The prevalence of the S/R genotype among CVD and CHD patients was not distinct from that in the control groups. We conclude that the S/R polymorphism of GPIb alpha, flanking the initiator codon of the receptor, does not seem to be associated with surface levels of the protein, and is not an independent risk factor for arterial thrombosis. PMID- 10669150 TI - Impaired procoagulant-anticoagulant balance during hormone replacement therapy? A randomised, placebo-controlled 12-week study. AB - In this randomised, placebo-controlled 12-week study, sixty healthy postmenopausal women received either placebo (N = 16) or daily 2 mg micronised oestradiol, either unopposed (N = 16, E2 group) or combined with a progestagen for 14 days of each cycle (N = 28, E2+P group). RESULTS: As compared to placebo, plasma levels of AT III were reduced only in the E2 group (approximately 28%), plasma levels of protein C decreased only in the E2+P group (approximately 4%) and plasma levels of protein S decreased in both the E2 and E2+P group (approximately 21%). In both the E2 and E2+P groups, the plasma levels of factor VII (antigen and activity) showed a borderline significant increase (approximately 10%), whereas no significant change was observed in active factor VII. Plasma levels of tissue-type plasminogen activator (approximately 22%), urokinase plasminogen activator (approximately 25%) and plasminogen activator inhibitor type-1 (approximately 43%) decreased in the E2 and E2+P groups, whereas those of plasminogen increased (approximately 12%). Treatment was associated with an increase in levels of prothrombin fragment 1+2 (approximately 31%), but levels of thrombin-antithrombin III complexes, and of plasmin-alpha2-antiplasmin complexes and total fibrin(ogen) degradation products did not change significantly. CONCLUSION: Short-term E2 and E2+P treatment is associated with a shift in the procoagulant-anticoagulant balance towards a procoagulant state. A substantial proportion of women do not have a net increase in fibrinolytic activity. These data may be relevant in explaining the increased risk of venous thromboembolism associated with ERT and HRT, and possibly also in explaining the negative results of the Heart and Estrogen/progestin Replacement Study. PMID- 10669151 TI - Inherited macrothrombocytopenia with distinctive platelet ultrastructural and functional features. AB - We report a family with inherited macrothrombocytopenia and characteristic large membrane complexes in the platelets. Two affected subjects had platelet counts of 40 and 65 x 10(9)/L respectively as assessed by contrast phase microscopy. Ultrastructural studies revealed giant spheroid platelets with characteristic large membrane complexes and/or giant vacuoles containing platelet organelles. Immunohistochemical studies of actin and tubulin showed a disorganization of the microtubule and actin systems. These abnormalities were absent in leukocytes, indicating a platelet-specific cytoskeleton disorder. Platelet autoantibodies were repeatedly absent. Nevertheless, in the peripheral blood we observed several figures of platelet phagocytosis by macrophages and neutrophils. The in vitro aggregometric response of platelets to ADP, collagen, thrombin, ristocetin was present, but shape change was absent. The urinary excretion of thromboxane A2 metabolites of the affected subjects were approximately 2 standard deviations above control values, in spite of a reduced maximal biosynthetic capacity of thromboxane from giant platelets assessed in vitro during whole blood clotting. This inherited platelet disorder shows structural and functional features which allow to distinguish it from other syndromes associated with giant platelets. We also propose to include ultrastructural and cytoskeletal studies in the diagnosis as well as in the classification of inherited giant platelet disorders. PMID- 10669152 TI - Screening for autoimmune markers is unnecessary during follow-up of adults with autoimmune thrombocytopenic purpura and no autoimmune markers at onset. AB - In an attempt to evaluate the frequency of autoimmune markers in autoimmune thrombocytopenic purpura (AITP) and to determine if autoimmune markers in patients with isolated AITP were associated with particular disease manifestations, we analyzed records of 122 consecutive adults with AITP. Twenty nine patients (24%) had significant titers of one or several autoimmune markers at AITP onset. Among them, 16 (13%) had antinuclear antibodies. The presence of autoimmune markers did not correlate with presenting feature, response to treatment or long-term outcome of AITP. Six patients (5%) developed seven autoimmune diseases during follow-up, comprising systemic lupus erythematosus, an antiphospholipid syndrome, autoimmune haemolytic anemia (n = 2), Grave's disease, Hashimoto's disease and primary biliary cirrhosis. At AITP onset, three of these patients had isolated biological markers of the autoimmune disease they later developed. The annual average incidence rate of autoimmune diseases was 1% per patient-year in the entire group and 0.4% in the group of patients with no autoimmune markers at AITP onset. This low rate is probably due to careful assessment at diagnosis for concomitant overt autoimmune disease. We recommend extensive screening for autoimmune markers at AITP onset, and careful follow-up of patients with autoimmune markers. Routine screening for autoimmune markers during AITP follow-up is not necessary for patients with no autoimmune markers at AITP onset. Systemic lupus erythematosus (SLE) and other autoimmune disorders can complicate autoimmune thrombocytopenic purpura (AITP) or be diagnosed concomitantly with otherwise unremarkable AITP (1, 2). However, the frequency and prognostic value of isolated autoimmune markers (i.e. not associated with an autoimmune disorder), particularly antinuclear antibodies (ANA) at AITP onset or during follow-up is controversial (3-8). For example, the committee organized by George et al. (9) to write guideline on the diagnosis and treatment of AITP stated that the search for ANA and lupus anticoagulant were of "uncertain appropriateness at diagnosis and during follow-up". In an attempt to help practicians to make decisions, we analyzed the frequency of autoimmune markers and autoimmune disorders at onset and during the follow-up in 122 adults with AITP and no overt autoimmune disease at diagnosis. These consecutive patients were followed by the same physician for a mean period of 6 years, and had routine screening tests for autoimmune markers and disorders at onset, before steroid therapy, and regularly during follow-up. PMID- 10669153 TI - Fluvastatin decreases soluble thrombomodulin in cardiac transplant recipients. AB - We conducted a randomized, placebo controlled, double-blind, cross-over study, to assess the effects of a 4-week fluvastatin therapy on plasma markers of endothelial activation or injury in 20 transplanted heart recipients. The levels of thrombomodulin and von Willebrand factor antigen were higher at baseline in cardiac transplant recipients than in age and sex-matched healthy controls. Plasma total cholesterol showed a 21% reduction on fluvastatin therapy (p = 0.0001). Fluvastatin treatment had no significant effect on creatininemia, plasma cyclosporine, PAI-1 antigen, PAI-1 activity, tPA antigen, and Von Willebrand factor. However, fluvastatin produced a significant decrease of plasma thrombomodulin (66.7 ng/ml on placebo versus 58.8 ng/ml on fluvastatin, p <0.001), suggesting a rapid improvement of endothelial injury in these patients. PMID- 10669154 TI - The management of oral anticoagulant therapy: the patient's point of view. AB - The aims of this study were to investigate on the general adhesion of the patients to oral anticoagulant therapy, and particularly on the quality of life of our patients, the doctor-patient relationship and the Centre-patient relationship. For this purpose we administered a questionnaire containing 17 main questions each with a maximum of 4 secondary questions. The questionnaire was administered to two groups of 127 and 137 oral anticoagulated patients (127 males and 137 females, mean age 55 +/- 19 years), followed at two Anticoagulation Clinics, in two Italian cities, Cagliari (Sardinia) and Padua (North East Italy). The cities differed in the number of patients monitored and the management modalities of anticoagulation. The results show that oral anticoagulant therapy does not limit the life-style of the patients. Only 11% of the patients complain of limitations to their daily life. Fifty-two percent believe their health has improved, and 87% are not afraid of negative consequences. The doctor-patient relationship is considered very important by 96% of patients. Seventy-eight percent refer to the Anticoagulation Clinic also for other health problems, 93% consider it important to be assessed by the doctor at the Anticoagulation Clinic, while 83% believe the doctor should always hand out the results personally. We conclude that in general oral anticoagulant therapy is accepted by the majority of patients, in spite of the need for periodic monitoring. The doctor-patient relationship should be taken into account, even in the case of a monitored, computer-assisted method of dose-adjustment. PMID- 10669155 TI - Hemostatic parameters and platelet activation marker expression in cyanotic and acyanotic pediatric patients undergoing cardiac surgery in the presence of tranexamic acid. AB - We have investigated hemostatic parameters including platelet activation in 56 pediatric patients with or without cyanosis undergoing cardiopulmonary bypass (CPB) and cardiac surgery to repair congenital defects. Patients were participants in a study assessing the effects of tranexamic acid on surgery related blood loss. Parameters monitored included blood loss, prothrombin F1.2, thrombin-antithrombin complexes, t-PA, PAI-1, plasminogen, fibrin D-dimer, and plasma factor XIII. Additionally, flow cytometry monitored platelet degranulation (P-selectin or CD63), as well as surface-bound fibrinogen, von Willebrand factor and factor XIIIa. Cyanotic patients had evidence of supranormal coagulation activation as both fibrin D-dimer and PAI-1 levels were elevated prior to surgery. While the extent of expression of P-selectin or CD63 was not informative, platelet-associated factor XIIIa was elevated in cyanotic patients at baseline. In both patient groups, CPB altered platelet activation state and coagulation status irrespective of the use of tranexamic acid. PMID- 10669156 TI - Purification and characterization of factor VII inhibitor found in a patient with life threatening bleeding. AB - We recently observed a patient with acquired inhibitor-induced F.VII deficiency whose plasma level of F.VII was < 1.0%. However, the biochemical nature of the inhibitor has not yet been clarified. In the present study, we purified the F.VII inhibitor from the patient's plasma by using activated F.VII (F.VIIa)-conjugated gel and characterized the inhibitor. The results showed that the inhibitor comprised two kinds of antibodies: one was eluted with EDTA (antibody 1) and the other with glycine-HCl buffer (pH 2.3) (antibody 2) from the F.VIIa affinity gel. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting analysis of these inhibitors demonstrated that both antibodies had features of immunoglobulin G1 (IgG1) with kappa and lambda-light chains. Antibody 1 bound to the immobilized F.VIIa with a high affinity in the presence of calcium ion, while antibody 2 bound to the F.VIIa very weakly and the binding was independent of calcium ion. Immunoblotting analysis demonstrated that antibody 1 bound to the light chain of F.VIIa after reduction with 2-mercaptoethanol, while it did not react with either the gamma-carboxyglutamic acid (Gla)-domainless light chain of F.VIIa or the heavy chain with the protease domain. Antibody 1 markedly inhibited the activity of tissue factor-F.VIIa complex. Based on these observations, it is suggested that F.VIIa autoantibody (antibody 1) recognizes the calcium-dependent conformation within or near the Gla domain and inhibits F.VIIa activity by interacting with the light chain. PMID- 10669157 TI - The half-life of infused factor VIII is shorter in hemophiliac patients with blood group O than in those with blood group A. AB - A considerable inter-individual variation in half-life of infused factor VIII is observed among patients with hemophilia A. The factors contributing to this wide range in factor VIII half-life are not known in detail. We analysed the pharmacokinetics of infused factor VIII in 32 patients with hemophilia A, comprising 20 brothers from 10 families, 3 and 4 brothers from 2 families, and 5 patients from 5 single families, respectively. Multiple linear regression analysis was used to assess the effect of several variables on factor VIII half life. We found that the pre-infusion von Willebrand factor antigen levels (vWF:Ag) were positively correlated with factor VIII half-life (r = 0.52, p = 0.002), i.e., each variable was associated with about 27% of the variance of the other. In fraternal pairs, familial clustering was significant for ABO blood group (p < 0.001), but could not be detected for factor VIII half-lives or pre infusion vWF:Ag levels. vWF:Ag level (p = 0.001) and ABO blood group (p = 0.003) significantly determined factor VIII half-life, whereas age, length, bodyweight, the presence or absence of a factor VIII gene inversion, and Rhesus phenotype did not. Patients with blood group O exhibited a statistically significant shorter factor VIII half-life than patients with blood group A (15.3 versus 19.7 h, respectively) (p = 0.003). Patients with blood group A and O differ in respect to the presence of anti-A antibodies in the latter. It is possible that these anti-A antibodies interact with endogenous vWF, thus affecting the half-life time of the factor VIII/vWF complex. PMID- 10669158 TI - Rescue of fatal neonatal hemorrhage in factor V deficient mice by low level transgene expression. AB - Factor V (FV) is a critical component of the coagulation cascade. FV-deficient patients suffer moderate to severe bleeding, though residual FV activity is detectable in nearly all cases. In contrast, FV-deficient mice die either during mid-embryogenesis, or of massive perinatal hemorrhage. In order to examine the requirements for FV in murine embryogenesis and hemostasis, we generated transgenic mouse lines expressing a Fv minigene under control of either the tissue-specific albumin (Malb) or rat platelet factor 4 (Rpf4) promoter. A total of 12 Malb and 3 Rpf4 lines were analyzed. Though expression in the target tissue was detectable in most lines by RT-PCR, only low levels of transgene expression were achieved (<3% of endogenous Fv in all lines). Despite a low level of Fv transgene expression, rescue of the lethal Fv-/- phenotype was observed with one of the Malb transgenic (Tg+) lines. However, rescue appeared to be incomplete with continued loss of >1/2 of expected Tg+,Fv-/- mice in early embryogenesis. Rescued Tg+,Fv-/- mice have undetectable FV (<0.1%) in both plasma and platelet compartments, but survive the perinatal period and mature to adulthood without spontaneous hemorrhage. We conclude that FV present at <0.1% is sufficient to support postnatal survival. Failure of the Malb transgene to rescue the midembryonic block suggests that FV expression is required during mammalian development at higher levels or with a different tissue-specific or temporal pattern. Taken together, these data may explain the observation of residual FV activity in most human FV-deficient patients due to early embryonic lethality in those absolutely deficient, and suggest that minimal levels of FV expression, below the level of detection, also may be sufficient to support survival in humans. PMID- 10669159 TI - Structural basis for hemophilia A caused by mutations in the C domains of blood coagulation factor VIII. AB - Three dimensional homology models for the C1 and C2 domains of factor VIII (FVIII) were generated. Each C domain formed a beta-sandwich, and C1 was covalently connected to C2 in a head-to-head orientation. Of the >250 missense mutations that cause FVIII deficiency and hemophilia A, 34 are in the C domains. We used the FVIII C1-C2 model to infer the structural basis for the pathologic effects of these mutations. The mutated residues were divided into four categories: 15 conserved buried residues that affect normal packing of the hydrophobic side chains, 2 non-conserved buried residues that affect structure, 11 conserved exposed residues and 6 non-conserved exposed residues. The effects of all 34 missense mutations can be rationalized by predictable disruptions of FVIII structure while at most four mutations (S2069F, T2154I, R2209Q/G/L and E2181D) may affect residues directly involved in intermolecular interactions of FVIII/VIIIa with other coagulation factors or vWF. PMID- 10669160 TI - Thirty-three novel mutations in the protein C gene. French INSERM network on molecular abnormalities responsible for protein C and protein S. AB - We analyzed the protein C gene (PROC) with the denaturing gradient gel electrophoresis (DGGE) scanning strategy in a series of 129 patients with suspected protein C (PC) deficiency (93 with low plasma PC levels and 36 with borderline level). At least one sequence variation was found in 104 of the 129 patients. Thirty-nine sequence variations (found in 72 patients) were already reported detrimental mutations. Thirty-three were novel sequence variations, of which 19 (found in 25 patients) were probably detrimental. Five novel mutations (A1T, R9H, S11R, S12R and K193Q) were associated with qualitative plasma PC deficiency, suggesting or confirming the functional importance of amino acids at these positions. This strategy confirmed the diagnosis of inherited PC deficiency in 79/93 (84.9%) patients with low plasma PC levels and 14/36 (38.8%) patients with borderline values. In order to explain abnormal PC levels observed in patients who did not carry detrimental mutations, screening for the -1654C/T and 1641A/G PROC promoter polymorphisms known to influence plasma PC concentrations was performed. The frequency of the CG allele associated with lower PC concentrations was slightly but not significantly lower in 82 heterozygotes for detrimental PROC gene mutations than in 36 patients with no identified detrimental mutations. PMID- 10669161 TI - Characterization of transgenic mice that secrete functional human protein C inhibitor into the circulation. AB - Protein C inhibitor (PCI) is a heparin binding serine protease inhibitor in plasma, which exerts procoagulant activity by inhibiting thrombomodulin-bound thrombin or activated protein C (APC). Since the role of PCI in vivo is largely unknown we generated genetically modified mice with expression of human PCI mRNA in hepatocytes only. Three transgenic lines have been characterized. Transgenic mice did not show gross developmental abnormalities. Two lines showed a pericentral and one line showed a periportal expression pattern of human PCI mRNA in the liver. Genetically modified mice secreted a functional transgenic protein into the circulation (3-5 microg/ml plasma in heterozygous mice and 10 microg/ml in homozygous mice), which inhibited human APC activity in the presence of heparin. Interestingly, transgenic mice in which human PCI was expressed periportally in the liver had the highest specific activity. Endogenous mouse PCI mRNA could only be detected in the male and female reproductive system, but not in the liver, indicating that endogenous PCI levels in the circulation are low or even absent in mice. These results demonstrate that the human PCI transgenic mice are a suitable model for studying the in vivo role of PCI in blood coagulation. PMID- 10669163 TI - A reliable and reproducible ELISA method to measure ristocetin cofactor activity of von Willebrand factor. AB - The ristocetin induced binding of vWF to GPIb, which is routinely tested in a platelet agglutination assay, can be reproducibly studied in an ELISA where plasma vWF binds to a captured rGPIb alpha-fragment (His1-Val289) in the presence of ristocetin. This binding is specific since the vWF-GPIb interaction could (i) be blocked by inhibitory anti-GPIb or anti-(vWF A1 domain) monoclonal antibodies (mAbs) and (ii) be enhanced by an anti-vWF mAb that also facilitates ristocetin induced platelet agglutination. Further studies were undertaken to determine whether the test could be used to differentiate vWF from patients with different types of von Willebrand's disease. The median vWF:RiCof activity in controls (n = 24) was 0.75 U/ml, in type 1 vWD patients (n = 17) 0.28 U/ml, in type 2A (n = 18) 0.055 U/ml, in type 2B (n = 4) 0.094 U/ml and in type 3 (n = 3) <0.0005 U/ml. Moreover, the values correlated well with those obtained from the vWF:RiCof agglutination assay (r = 0.873). The vWF:RiCof-ELISA has several advantages: the use of a recombinant fragment instead of donor platelets results in a more reproducible test with a low inter- and intra-assay variability (<14% CV), the test can further be readily automated and for a single determination, only minimal amounts of patient plasma are required (8 microl). PMID- 10669162 TI - Homozygosity for the protein S Heerlen allele is associated with type I PS deficiency in a thrombophilic pedigree with multiple risk factors. AB - The multifactorial character of thrombotic disease is shown in a Spanish pedigree in which the propositus, with recurrent deep vein thrombosis, inherited the factor V R/Q506 mutation, the prothrombin 20210G/A variant and type III Protein S deficiency. Among 14 relatives carrying one or two of these three risk factors, thrombosis is present in a heterozygote for R/Q506 and in another for 20210G/A, who also had slightly positive antiphospholipid antibodies. Type I PS deficiency was also found in a young asymptomatic woman. PROS1 analysis showed coexistence of type III and type I PS deficiency to be associated with heterozygosity and homozygosity, respectively, for the P460 or PS Heerlen allele of the S/P460 variant. Analysis of PS values in this and other pedigrees segregating this variant revealed that not only free but also mean total PS levels are slightly but significantly lower in the SP460 heterozygotes than in the SS460 homozygotes. These findings strongly suggest a role of the P460 variant in the expression of the PS deficient phenotype. PMID- 10669164 TI - Induction of tissue factor procoagulant activity in myelomonocytic cells inoculated by the agent of human granulocytic ehrlichiosis. AB - Human granulocytic ehrlichiosis (HGE) is a recently recognized rickettsial tick borne febrile illness that may occasionally be complicated by coagulopathy. The agent of HGE (aHGE) is an obligate intracellular pathogen, which replicates in endosomes within neutrophils and their precursors. We hypothesized that aHGE might cause DIC via induction of monocyte tissue factor procoagulant activity (TF PCA). Peripheral blood mononuclear cells (PBMNC) and HL-60 cells were used to model the effect of aHGE infection on monocytes/macrophages. Mononuclear cells inoculated with aHGE in vitro demonstrated approximately a 12-15-fold increase in TF PCA, with peak activity occurring at 8-12 h. HL-60 cells inoculated with aHGE also manifested a 4-6 fold induction of TF PCA, with maximal activity occurring at about 8 h. By comparison, E. Coli lipopolysaccharide (LPS) also induced an increase in TF PCA of an equivalent magnitude, and with a similar time course. Induction of TF did not require inoculation of HL-60 cells with live organism, since heat-inactivated aHGE still stimulated TF PCA expression in the target cells. Furthermore, filtered supernatants from heat-inactivated organisms induced TF PCA suggesting that the effect is due to a soluble mediator produced by the organism. Although aHGE is a gram negative organism, the soluble mediator did not appear to be classic endotoxin in that the supernatants tested negative for endotoxin by the Limulus Amoebocyte assay, and polymixin had no inhibitory effect on aHGE supernatants. We conclude that aHGE induces cells of the myelo-monocytic lineage to synthesize TF, which may contribute to the clinical coagulopathy that can be observed in this condition. An atypical soluble mediator or cellular component of the organism appears to be critically important in TF induction by aHGE. PMID- 10669165 TI - Glycoprotein Ib-binding protein from the venom of Deinagkistrodon acutus--cDNA sequence, functional characterization, and three-dimensional modeling. AB - Agkicetin-C, a potent glycoprotein Ib antagonist from the venom of the Chinese pit viper, Deinagkistrodon acutus, has been purified and characterized (5). It is a disulfide-linked heterodimer containing subunits of 132 and of 123 amino acid residues. Herein, the complete amino acid sequences were resolved by cloning and nucleotide sequencing of the cDNAs. The sequences of its subunits are homologous to those of other snake venom proteins of the C-type (Ca2+-dependent) lectin superfamily. A three-dimensional model of agkicetin-C was constructed based on the crystal structure of habu coagulation factor IX/X-binding protein. By careful alignment of all the related sequences available and comparing the 3D-model of agkicetin-C with structures of other homologous proteins of different functions, some variable residues of agkicetin-C were identified, which possibly are responsible for the specificity of this distinct subtype of the C-type lectin like venom proteins. PMID- 10669166 TI - Collagen binding assay for von Willebrand factor (VWF:CBA): detection of von Willebrands Disease (VWD), and discrimination of VWD subtypes, depends on collagen source. AB - A large number of different collagen preparations [n = 21] have been assessed for their ability to both detect von Willebrands Disease (VWD), and discriminate different VWD subtypes. Collagen preparations were tested at a range of concentrations and included: Type I, III and IV, and various mixtures of these, as aqueous supplied preparations and/or reconstituted from bulk lyophilised stock. Tissue sources for collagens ranged from human placenta to calf skin to equine tendon. Three of the collagen preparations tested did not support von Willebrand factor (VWF) binding in an ELISA process (therefore unable to detect VWD). The ability of the remaining preparations to detect VWF was variable, as was their ability to discriminate VWD subtypes. Detection of VWF and discrimination of VWD subtypes was not mutually inclusive. Thus, some collagen preparations provided excellent detection systems for VWF, but comparatively poorer discrimination of Type 2 VWD, while others provided good to acceptable detection and discrimination. Subtype discrimination was also dependent on the collagen concentration, and some batch to batch variation was evident with some preparations (particularly Type I collagens). Overall, best discrimination was typically achieved with Type I/III collagen mixtures, or Type III collagen preparations (where effectiveness was highly dependent on concentration). Good discrimination was also achieved with a commercial Type III collagen based VWF:CBA kit method. Results of the various 'VWF:CBA assays' are also compared with those using the Ristocetin Cofactor (VWF:RCof) assay (by platelet agglutination) and that using a commercial 'VWF:RCof-alternative/activity' ELISA procedure. These latter methodologies tended to be less sensitive to VWF discordance when compared to that detected by the majority of the VWF:CBA procedures. PMID- 10669167 TI - Von Willebrand Disease type 2M "Vicenza" in Italian and German patients: identification of the first candidate mutation (G3864A; R1205H) in 8 families. AB - Von Willebrand disease type 2M "Vicenza" (VWD 2M V) is characterised by autosomal dominant inheritance, low von Willebrand factor (VWF) and the presence of "supranormal" multimers in plasma. This specific phenotype has been described in Italian and recently also in German patients. The molecular defect is linked to the VWF gene. However, no specific mutations have been identified until now. We analysed the complete coding region and adjacent intron sequences of the VWF gene in Italian families in comparison to German families with VWD 2M V by a PCR-based mutation screening, combined with SSC- and heteroduplex-analysis of exons 2 through 52, followed by direct sequencing. We identified the first heterozygous candidate mutation (G3864A; R1205H) in all affected members of the 7 Italian families and in 1 German patient but not in the unaffected family members nor on 100 chromosomes of normal subjects, suggesting a causal relationship between the mutation and the phenotype. Haplotype identity, with minor deviations in one Italian family, suggests a common but not very recent genetic origin of R1205H. PMID- 10669168 TI - Nucleotide sequence of the gene encoding murine tissue factor pathway inhibitor 2. AB - Tissue factor pathway inhibitor-2 (TFPI-2), also known as placental protein 5, is a 32 kDa extracellular matrix-associated serine proteinase inhibitor consisting of three tandemly-arranged Kunitz-type domains. Two overlapping genomic clones containing sequences encoding murine TFPI-2 were isolated from a lambda FIXII 129 SVJ mouse genomic library, and the complete nucleotide sequence of the gene was determined. The murine TFPI-2 gene spans approximately 9.3 kilobases and consists of five exons and four introns. The nucleotide sequences surrounding all the exon intron boundaries are highly conserved and obey the GT-AG rule. Each Kunitz-type domain is encoded by a single exon, similar to that observed for other Kunitz type proteinase inhibitors. A total of 1,577 bp of the 3'-flanking region contains a probable polyadenylation site (ATTAAA) at +5,759 and an apparent cleavage or termination site (CATTG) at +6,170. The 5'-flanking region of the murine TFPI-2 gene contains a prototypical TATA box, a GC box and two CAAT boxes. In addition, several candidate transcription factor binding sites responsible for placenta-, endothelial cell-, and smooth muscle cell-specific expression of the TFPI-2 gene were also identified. PMID- 10669169 TI - The effect of monoclonal anti-human-platelet antibodies on platelet kinetics in a baboon model: IgG subclass dependency. AB - We assessed the in vivo effect of six intact anti-human antiplatelet antibodies of two major IgG subclasses on platelet kinetics in baboons. Five of the six antibodies tested caused thrombocytopenia of varying degree when injected at a precalculated threshold value. An agglutinating IgG1 antibody (MA-8L4A12) caused a long-lasting, mild thrombocytopenia with a predominant uptake of radiolabelled platelets in the spleen, while the four IgG2 antibodies tested (MA-13G8E1, MA 2M5A6, MA-21K2E8 and MA-22M10) caused a severe, transient thrombocytopenia with uptake of platelets in the liver. Two of the IgG2 antibodies (MA-13G8E1 and MA 2M5A6) caused platelet activation and aggregation in vitro, whilst the other two did not elicit a platelet aggregation response. The platelet survival time was shortened with all five of the thrombocytopenia-inducing antibodies, while only one antibody (MA-2M5A6) had a significant effect on the bleeding time. This study indicates that the IgG subclasss may be a determining factor in the outcome of platelet sequestration in immune-induced thrombocytopenia. PMID- 10669170 TI - Platelets release their lysosomal content in vivo in humans upon activation. AB - Platelets contain, besides alpha- and delta-granules, lysosomes which store glycohydrolases able to degrade glycoproteins, glycolipids and glycosaminoglycans. While several studies have shown that alpha- and delta granule secretion takes place "in vivo" in humans upon platelet activation, no data are available on the "in vivo" release of lysosomes. We have studied the release of platelet lysosomal contents "in vivo" in healthy volunteers at a localized site of platelet activation by measuring markers of lysosomal secretion in the blood oozing from a skin wound inflicted for the measurement of the bleeding-time. The levels of beta-N-acetylhexosaminidase (Hex) were 13.1 +/- 0.85 mU/ml in bleeding-time blood and 10.2 +/- 0.66 mU/ml in plasma (p <0.001). Hex in serum was 16.4 +/- 0.72 mU/ml. The levels of beta-galactosidase were also higher in bleeding-time blood than in plasma (0.85 +/- 0.07 mU/ml vs 0.4 +/- 0.05 mU/ml, p <0.001). In bleeding-time blood collected at one minute intervals, Hex rose progressively consistent with ongoing platelet activation and flow-cytometry showed a progressive increase of the expression of LIMP and LAMP-2, two lysosomal associated proteins. In conclusion, our data demonstrate that platelet lysosomal glycohydrolases are released "in vivo" in humans upon platelet activation. PMID- 10669171 TI - Determination of the putative binding sites for thrombin receptor activating peptide through a hydropathic complementary approach. AB - Putative binding sites in a platelet thrombin receptor (PAR-1) for the activating peptide SFLLRNPNDKYEPF (AP) and for the bradykinin analogue MKRPPGFSPFRSSRIG were revealed using a computer program for identifying complementary peptide segments. The program is based on the assumption that interactions of agonist's peptides and protein's receptors can be elucidated by complementary average hydropathies as much as possible equal by size and opposite by sign. Some of the computer found putative binding sites were close to the supposed AP-PAR-1 contacts in the amino-terminal exodomain and in the second extracellulary loop of PAR-1. Peptides complementary to these binding sites were also computer-designed and were synthesized. They mostly inhibited the aggregation of gel filtered platelets by thrombin (0.025 U/mL) with IC50 in a high micromolar range of concentrations. The peptide complementary to site L258-Y269 of PAR-1 induced aggregation of gel filtered platelets with EC50 = 98 [micromol/L] related to thrombin (0.025 U/mL) aggregation response. PMID- 10669172 TI - Prevention of deep vein thrombosis after hip replacement. PMID- 10669173 TI - Further evidence that activated protein C resistance affects protein C coagulant activity assays. PMID- 10669174 TI - Is there a role for anti-phospholipid-binding protein antibodies in the pathogenesis of thrombosis in Behcet's disease? PMID- 10669175 TI - Transient lupus anticoagulants in children: stepwise disappearance of diagnostic features. PMID- 10669176 TI - GPIaIIa as a candidate target for anti-platelet autoantibody occurring during valproate therapy and associated with peroperative bleeding. PMID- 10669177 TI - Agreement of D-dimer results measured by a rapid ELISA (VIDAS) before and after storage during 24h or transportation of the original whole blood samples. PMID- 10669178 TI - Genotype distribution of the 46C/T polymorphism of coagulation factor XII in the Japanese population: absence of its association with ischemic cerebrovascular disease. PMID- 10669179 TI - Factors affecting prescribing of the newer antidepressants. AB - OBJECTIVE: To survey various prescriber types and specialties to determine whether differences exist in prescribing patterns for the newer antidepressants. DESIGN, SETTING, AND PARTICIPANTS: A survey about prescribing of the newer antidepressants was mailed to 1,500 New York state licensed prescribers who were randomly selected from membership rosters. Nurse practitioners; physician assistants and physicians in family medicine, primary care, general practice, and internal medicine; and psychiatrists were included. MAIN OUTCOME MEASURES: Prescriber responses regarding factors involved with choosing among the newer antidepressants. RESULTS: A total of 508 surveys (36%) were returned, of which 398 (29%) were acceptable for analysis. In choosing among the newer antidepressants, most prescribers ranked patient diagnosis and past success as a high priority, and free drug samples and drug-representative detailing as a low priority. The majority of each prescriber type preferred fluoxetine for major depression and depression associated with fatigue; paroxetine for concomitant anxiety and depression, as well as for panic disorder; and sertraline for geriatric patients and patients with suicidal ideation. Differences existed between the prescriber groups when asked whether prescribing habits for the newer antidepressants were based on familiarity with a particular agent (p = 0.0009) and on labeled indications (p = 0.002). CONCLUSIONS: This is the first study to demonstrate prescribing preferences for the newer antidepressants among different prescriber groups. Additional studies are needed to determine predictors of patient response to newer antidepressants and clinical guidelines for their use. PMID- 10669180 TI - Adverse effects of reduced-dose d-penicillamine in children with mild-to-moderate lead poisoning. AB - BACKGROUND: Oral chelation therapy with d-penicillamine (d-PCN) has been proven to be effective in the treatment of mild-to-moderate lead poisoning. However, d PCN is associated with a relatively high incidence of adverse effects when given in the standard dose of 25-30 mg/kg/d. Lower doses of d-PCN may reduce the rate of adverse effects without a significant reduction in the drug's efficacy. OBJECTIVE: To examine the incidence of rash, white blood cell and platelet count depression, and abnormal urinalysis with d-PCN when given in a dose of 15 mg/kg/d to children with blood lead concentrations <40 microg/dL. METHODS: Retrospective analysis of a clinical treatment course of children who received d-PCN during 1996 in the Lead and Toxicology Clinic of Children's Hospital, Boston. All children were treated under a reduced-dose d-PCN chelation protocol. RESULTS: During the study period, 55 children (mean age 37.4 mo) received 66 courses of d PCN. Mean blood lead concentration before chelation was 24 microg/dL (range 15 37), with a corresponding erythrocyte protoporphyrin concentration of 42 microg/dL. After 77 days of treatment with d-PCN, blood lead concentration was reduced to mean 16 microg/dL (mean fall 35%; p = 0.005) and erythrocyte protoporphyrin was reduced to 28 microg/dL (p = 0.009). During chelation therapy, the white blood cell count fell below 5,000/mm3 in seven cases (9.7%); there were no episodes of platelet counts falling below 150,000/mm3. No cases of abnormal urinalysis were reported; three episodes of rash (4.5%) were recorded. The only patients prematurely terminated from therapy were those who developed rash; in all three cases, drug eruption was an isolated occurrence, which resolved within 48 hours of diphenhydramine therapy. All adverse effects were transient and resolved during or immediately after chelation therapy. CONCLUSIONS: Reduced-dose d-PCN appears to maintain efficacy at reducing blood lead concentrations. Reduced dose d-PCN also appears to be associated with a rate of adverse effects lower than previously reported; observed adverse effects appear to be benign and transient. PMID- 10669181 TI - Treatment of uncomplicated urinary tract infections: exploring differences in adherence to guidelines between three European countries. Drug Education Project Group. AB - OBJECTIVE: To evaluate adherence of general practitioners to treatment guidelines regarding urinary tract infections in three European countries and to investigate whether differences in adherence at the prescribing level within and between countries could be explained by general practitioners' knowledge and attitudes, characteristics, or national setting. DESIGN: Prescribing data collected in 1994 1995 were analyzed regarding use of first-choice drugs and duration of treatment, knowledge and attitudes were assessed with a questionnaire, and multiple regression analysis was used to explain differences in prescribing behavior within and between countries. RESULTS: Our study is based on data from 85.6% of the 584 general practitioners who were scheduled to participate in a continuing education program. The mean proportion of responses in agreement with the guidelines regarding first-choice drugs was 0.69 in Sweden, 0.78 in the Netherlands, and 0.79 in Norway; regarding duration of treatment, the mean proportion was 0.56 in Sweden, 0.67 in the Netherlands, and 0.59 in Norway. The proportion of first-choice drugs prescribed for women (18-75 y) was 0.55 in Sweden, 0.83 in the Netherlands, and 1.00 in Norway (patients >16 y). The duration of treatment was 7.6 defined daily doses per prescription in Sweden, 5.9 in the Netherlands, and 6.6 in Norway. Knowledge and attitudes explained 0-17% of the variation in prescribing. Years in practice explained 0-11%, and the general practitioners' gender had no explanatory value. The national setting explained most of the variation between countries. CONCLUSIONS: Differences in prescribing behavior can be explained only to a small extent by deviations from the guidelines in terms of knowledge and attitudes. Between countries, differences in regulation, marketing, and distribution of drugs seem to be of much greater importance. PMID- 10669182 TI - Renal allograft dysfunction associated with rifampin-tacrolimus interaction. AB - OBJECTIVE: To report an interaction between tacrolimus and rifampin with subsequent adverse effects on renal allograft function. CASE SUMMARY: A 61-year old Chinese man received a cadaveric renal transplant in 1991. Progressive deterioration of allograft function developed during the following six years while the patient was receiving cyclosporine and prednisolone. In January 1998, tacrolimus was substituted for cyclosporine for late biopsy-proven graft rejection, with target trough blood concentrations between 5 and 8 ng/mL. After conversion, serum creatinine fell to 2.0 mg/dL; the nadir was reached within one year. At the same time, rifampin was instituted for controlling tuberculosis and empiric fluconazole was discontinued. Twelve days later, the patient's serum creatinine concentration rose to 2.9 mg/dL and tacrolimus concentration fell to 1.5 ng/mL, along with oliguria. These findings suggested acute rejection, which was successfully reversed by steroid therapy. However, more than a tenfold increase in the tacrolimus dosage was required to maintain the same concentrations during subsequent months, accompanied by an increase in serum creatinine (from 2.0 to 2.6 mg/dL) and decrease in urine excretion. Biopsy at this time demonstrated acute rejection (Banff I), chronic allograft nephropathy (Banff II), and suspected tacrolimus nephrotoxicity. After unsuccessful methylprednisolone recycling, mycophenolate mofetil was introduced to control rejection and facilitate reduction of the tacrolimus dosage to minimize its nephrotoxicity. CONCLUSIONS: As a potent CYP3A4 isoenzyme inducer, rifampin coadministration caused the abrupt decrease in tacroiimus blood concentrations, leading to an approximate tenfold increase in its daily dose, which may be important to subsequent allograft dysfunctions. PMID- 10669183 TI - Toxic epidermal necrolysis associated with acetaminophen ingestion. AB - OBJECTIVE: To report a case of toxic epidermal necrolysis (TEN) associated with acetaminophen ingestion. SUMMARY: A seven-year-old girl developed TEN after acetaminophen ingestion. The diagnosis was based on clinical evaluation and skin biopsy. A later acetaminophen challenge, undertaken by an allergist who questioned the diagnosis, resulted in a similar skin reaction. DISCUSSION: TEN is a severe disease with a high mortality rate. TEN may be either idiopathic or associated with several clinical conditions, such as viral infections, autoimmune disorders, malignancy, and drug hypersensitivity. Because of the rarity of its association with acetaminophen, the diagnosis in our patient was questioned by an allergist who performed an oral acetaminophen rechallenge test despite the potential risk. This caused a severe skin reaction that required rehospitalization. CONCLUSIONS: TEN can be caused by over-the-counter medications such as acetaminophen. Rechallenge with the causative drug carries a risk of severe complications and should be avoided. PMID- 10669184 TI - Fluorouracil-induced neurotoxicity. AB - OBJECTIVE: To report a case of acute neurologic adverse effects related to fluorouracil administration and to review the neurotoxicity of this agent. CASE SUMMARY: A 73-year-old white man with a history of esophageal carcinoma was treated with fluorouracil 1,500 mg iv daily for four days. After completing treatment, he presented with sudden onset of confusion, cognitive disturbances, a cerebellar syndrome, and repeated seizures. A magnetic resonance image of the brain showed no structural abnormalities, and cerebrospinal fluid examination was normal; none of the other laboratory tests provided an explanation for his symptoms. The patient was treated with anticonvulsants, and the cognitive changes resolved in 72 hours. The cerebellar signs, however, did not resolve completely and persisted when the patient was examined two weeks after discharge. DISCUSSION: Fluorouracil can cause both acute and delayed neurotoxicity. Acute neurotoxicity manifests as encephalopathy or as cerebellar syndrome; seizures, as seen in our patient, have rarely been reported. Acute neurotoxicity due to fluorouracil is dose related and generally self-limiting. Various mechanisms for such toxicity have been postulated, and treatment with thiamine has been recommended. Delayed neurotoxicity has been reported when fluorouracil was given in combination with levamisole; this form of subacute multifocal leukoencephalopathy is immune mediated and responds to treatment with corticosteroids. CONCLUSIONS: Clinicians should be aware of the adverse neurologic effects of fluorouracil and should include them in the differential diagnosis when patients receiving the drug present with neurologic problems. PMID- 10669185 TI - The effect of intravenous verapamil on cerebral hemodynamics in a migraine patient with hemiplegia. AB - OBJECTIVE: To describe the use of intravenous verapamil in a migraine patient with hemiplegia to reverse the symptomatology and hemodynamics of the middle cerebral artery as determined by transcranial Doppler. CASE SUMMARY: A 31-year old white woman was admitted with an acute exacerbation of migraine with hemiplegia. A transcranial Doppler showed an increased flow velocity through the middle cerebral artery consistent with a migrainous process. The patient was treated with verapamil 5 mg iv and the hemiplegia gradually resolved. A transcranial Doppler indicated that the flow velocity through the middle cerebral artery was decreased after verapamil administration, indicating reversal of the vasospasm. DISCUSSION: Transcranial Doppler has not been previously used to determine the effect of intravenous verapamil on the migrainous process. Intravenous verapamil reversed the altered hemodynamics of the middle cerebral artery as determined by transcranial Doppler. This finding correlated with the gradual resolution of hemiplegia. Whether both subjective and objective findings in this patient can be attributed to the reversal of the cerebral artery hemodynamics is not known. CONCLUSIONS: Intravenous verapamil appears to reverse the vasospasm that may be associated with a migrainous process. Whether this effect is solely responsible for clinical improvement is not known. Verapamil may be a consideration for the treatment of intractable migraine, especially when there is evidence of spasm of the major cerebral arteries. PMID- 10669186 TI - Dofetilide: a class III-specific antiarrhythmic agent. AB - OBJECTIVE: To review published reports on the pharmacology and clinical use of dofetilide in the management of cardiac dysrhythmias. DATA SOURCES: A MEDLINE search (January 1966-June 1999) was performed using dofetilide and UK-68,798 as key words. English-language articles were identified, and the references of these articles were used to further identify pertinent articles. STUDY SELECTION: All acquired studies and reviews discussing the pharmacology, pharmacokinetics, chemistry, and clinical efficacy of dofetilide were reviewed. DATA EXTRACTION: Articles were selected based on quality of review of the pharmacology and clinical use of dofetilide. Given the paucity of data on the clinical pharmacology and use of dofetilide, most articles obtained were used, including abstracts when full reports were not available. DATA SYNTHESIS: Dofetilide is a relatively specific class III antiarrhythmic agent. It increases action potential duration and effective refractory period without impacting conduction velocity. These actions of dofetilide are explained by its ability to inhibit the rapid component of the delayed, outward-rectifying potassium current, thus blocking the efflux of potassium during repolarization. Introductory investigations suggest that dofetilide may be of use in treating and preventing atrial dysrhythmias such as atrial fibrillation, atrial flutter, and paroxysmal supraventricular tachycardia. Dofetilide may also have a role in preventing ventricular tachycardia from occurring. Some data also suggest that dofetilide may improve the morbidity of heart failure patients. Currently, the most troublesome adverse effect of dofetilide is its propensity to induce ventricular proarrhythmias, especially torsade de pointes. CONCLUSIONS: Based on the data currently available, dofetilide should have a role in the pharmacotherapy of cardiac dysrhythmias, especially those of atrial origin. More data on its efficacy and tolerability are needed, however, to fully delineate dofetilide's role amid currently available antiarrhythmic agents. PMID- 10669187 TI - Homocysteine as a risk factor for atherosclerosis. AB - OBJECTIVE: To review the role of homocysteine as a risk factor in the pathogenesis of atherosclerosis and to provide recommendations for the treatment of hyperhomocysteinemia. DATA SOURCES: A MEDLINE search using key terms such as homocysteine, atherosclerosis, folic acid, vitamin B6, and vitamin B12 was conducted for the time period 1966 through January 1999. STUDY SELECTION: An article was selected for inclusion in this review if it assessed the relationship and proposed mechanisms of hyperhomocysteinemia on the vasculature, physiologic changes due to hyperhomocysteinemia, and outcomes due to hyperhomocysteinemia, such as morbidity and mortality. In addition, studies that assessed the treatment outcomes of hyperhomocysteinemia were evaluated. DATA SYNTHESIS: Studies of patients with cerebral vascular disease reveal elevated homocysteine concentrations in 30-40% of patients compared with controls. Many studies demonstrate a correlation between elevated homocysteine concentrations, risk of myocardial infarction, and mortality. In addition, hyperhomocysteinemia and decreased folic acid concentrations have been identified in end-stage renal disease (ESRD) and type 2 diabetic patients, while both concentrations remained normal in healthy controls. Studies using folic acid 650 microg/d reduced homocysteine concentrations to within normal therapeutic range after two weeks of treatment. Studies with vitamins B6 and B12 have demonstrated that the use of either alone is ineffective, but when combined or administered with folic acid, homocysteine concentrations return to normal. All therapies must be given for the lifetime of the patient. In addition, patients must use discretion in their diet, as common beverages, such as coffee, have a strong correlation with hyperhomocysteinemia, while foods high in folic acid, vitamin B6 and vitamin B12 may reduce homocysteine concentrations. Additional prospective studies are needed to determine effects of treatment of hyperhomocysteinemia and various diets on atherosclerotic morbidity and mortality. CONCLUSIONS: Studies demonstrate a positive correlation between hyperhomocysteinemia and atherosclerosis. The treatment of choice for hyperhomocysteinemia is folic acid. Although the optimal dose is not known, 650 microg/d is the minimum effective dose. To date, no studies have assessed the effects on morbidity and mortality when treating high homocysteine concentrations in atherosclerotic patients. PMID- 10669188 TI - Treatment of hypertension in the perioperative patient. AB - OBJECTIVE: To review studies and drug therapy relating to the treatment of hypertension in perioperative patients. DATA SOURCES: Articles were selected from a MEDLINE search (1966-August 1998), and several textbooks on hypertension and surgery were reviewed. In addition, bibliographies of all articles and textbook chapters were studied for articles not found in the computerized searches. STUDY SELECTION: Clinical studies involving hypertension in the perioperative setting were included. The initial search was limited to studies conducted in humans and published in English. DATA EXTRACTION: Information regarding drug therapy was reviewed and guidelines were constructed for managing surgical patients with acute blood pressure elevations. DATA SYNTHESIS: Although nitroprusside and nitroglycerin, with their short onset of action and duration of effect, are indicated for hypertensive emergencies, a variety of agents are available for hypertensive urgencies. An algorithm that can be used as a template for the development of intrainstitutional guidelines is provided. CONCLUSIONS: Due to the scarcity of comparative trials, decisions involving agents for the treatment of perioperative hypertension must often be made based on combined efficacy, toxicity, cost, and convenience considerations. PMID- 10669189 TI - Gender differences in depression and antidepressant pharmacokinetics and adverse events. AB - OBJECTIVE: To review data generated by studies examining gender differences in the prevalence of depression, as well as in antidepressant pharmacokinetics, pharmacodynamics, and adverse events. DATA SOURCES: Published articles and abstracts were identified through MEDLINE (January 1966-April 1999) using the following search terms: antidepressant, response, gender, pharmacokinetic, pharmacodynamic, female, side effect, and adverse events. All articles that assessed gender differences in antidepressant response, pharmacokinetics, and adverse events, as well as articles that evaluated postulated mechanisms for these differences, were reviewed. Additional articles were identified from bibliographies of retrieved articles. STUDY SELECTION AND DATA EXTRACTION: All relevant abstracts, studies, and review articles were evaluated. DATA SYNTHESIS: Gender differences in the prevalence of depression have been reported and may result from the interaction of several factors. Women have been shown to have a higher incidence of depression, which may be due to artifact, social, or biologic reasons. Studies suggest that the pharmacokinetic disposition of popular antidepressants varies between men and women, and women taking antidepressants may exhibit a different adverse event profile. Only one study specifically evaluated gender differences in antidepressant treatment response. CONCLUSIONS: Further research elucidating gender differences in response to antidepressant treatment and on depression prevalence is needed. Some studies report that the pharmacokinetics of antidepressants may vary between men and women. Therefore, clinicians should be aware that potential differences in antidepressant pharmacokinetics may exist, and a dosage adjustment may be necessary for women to ensure a favorable drug response, compliance, and decreased incidence of adverse events. PMID- 10669190 TI - Hydroxyurea in the treatment of HIV-1. AB - OBJECTIVE: To describe the role of hydroxyurea in the treatment of HIV-1. DATA SOURCES: Published clinical studies using hydroxyurea in HIV treatment were accessed through MEDLINE (January 1994-March 1999) and conference abstracts. All relevant studies were evaluated. DATA SYNTHESIS: Adherence, expense, and resistance limit the pharmacotherapeutic options in the management of patients with HIV. Hydroxyurea may be an alternative to conventional HIV treatments. CONCLUSIONS: Several potential advantages of adding hydroxyurea to antiretroviral treatment regimens include the drug's well-documented toxicity, convenient dosing, good tolerability and low cost, and its unique mechanism of action. Hydroxyurea may have synergistic effects that prove promising in initial and salvage therapy antiretroviral regimens. Larger, well-controlled clinical studies are needed to adequately define the role of hydroxyurea in the treatment of HIV. PMID- 10669191 TI - Amiloride for the prevention of amphotericin B-induced hypokalemia and hypomagnesemia. AB - OBJECTIVE: To review the published clinical data assessing the role of amiloride in the prevention of amphotericin B (AmB)induced electrolyte disorders. DATA SOURCES: A MEDLINE search (January 1966-April 1999) of English-language literature pertaining to AmB, amiloride, potassium, and magnesium was performed. Tertiary sources were also used. DATA EXTRACTION: In vivo and in vitro human and animal data and case reports were included due to the lack of published clinical trials. DATA SYNTHESIS: AmB administration can result in severe hypokalemia and hypomagnesemia requiring chronic supplementation. In one prospective, controlled study of hypokalemia with AmB administration, patients receiving concomitant amiloride had significantly greater potassium concentrations (p < 0.01) and required significantly less potassium supplementation (p < 0.001). Amiloride may also reduce the amount of magnesium supplementation required by sparing elimination through the kidneys. CONCLUSIONS: Amiloride may be considered for the prevention of AmB-induced hypokalemia and hypomagnesemia, especially in patients at high risk for complications resulting from these electrolyte disorders. Further studies are needed to assess concomitant use of other potassium-sparing diuretics and AmB. PMID- 10669192 TI - Sorbitol compared with xylitol in prevention of dental caries. AB - OBJECTIVE: To summarize published data on the comparative efficacy of sorbitol and xylitol for prevention of dental caries. DATA SOURCES: Published double-blind comparative trials, using sorbitol and xylitol products, identified by MEDLINE (January 1966-December 1998) and International Pharmaceutical Abstracts (January 1970-December 1998) searches. DATA SYNTHESIS: Clinical trials generally used sorbitol and xylitol gums, which patients chewed three to five times daily for 20 40 months. Xylitol was superior to sorbitol in two longer, secondary dentition trials (30-63% reductions), but not in two primary dentition trials. CONCLUSIONS: The data suggest that xylitol-containing gums may provide superior efficacy in reducing caries rates in high-risk populations. PMID- 10669193 TI - Medicine information help lines: a survey of hospital pharmacy-based services in the UK and their conformity with guidelines. AB - OBJECTIVE: To describe the prevalence and nature of hospital pharmacy-based medicine help lines for consumers in the UK and to compare service provision with published guidelines. BACKGROUND: Since 1992, telephone help lines for patients have proliferated in hospital pharmacies in the UK. There is no common template for such services with variations in target group, number and type of calls, and arrangements for training and audit. Data on these factors will help guide further development of such services. METHODS: All medicine help lines operating from hospital pharmacies in the UK were identified through the national Drug Information Pharmacists network. They were sent a piloted questionnaire covering many aspects of help line operation, including funding, method of advertisement, procedures, target group, number and nature of calls, and audit procedures. RESULTS: Eighty-two help lines were identified in England, Scotland, Wales, and Northern Ireland. Completed responses were received from 69 help lines (84% response rate). The pharmacy drug information center was the help line site in 57% of hospitals; all other help lines were located in the dispensary. In 55% of cases, help lines were open only to patients of the hospital. In the remainder of help lines, calls from the public were answered (although the majority of help lines only advertised to hospital patients). Calls were answered by pharmacists only in 45% of services, and additional staff training had been provided in 43%. Only 48% of services had written procedures or guidelines for operation of the help line. Forty-six percent of the services received fewer than five calls per week, 31% received between five and 10 calls per week, and 22% received 11 or more calls per week. In 59% of the sites, calls took an average of 10 minutes or less to answer; it took 11-15 minutes in 32% of the sites and >15 minutes in 9% of the sites. The most common queries related to adverse effects, dosage and administration, and interactions (including alcohol). Only 33% of help lines had any auditing or monitoring of the service in place. CONCLUSIONS: The increasing use of the telephone to provide services directly to consumers is reflected in the growth of hospital-based medicine help lines in the UK. The telephone route is likely to become more important as patients' needs for information about their medicines increase. However, the rate of calls is low when compared with the number of patients issued prescriptions; further research is needed to investigate the reasons for this low response. There is currently reason for concern because most help lines lack not only professional training in telephone counseling, but also proper documentation, monitoring, and audit procedures. PMID- 10669194 TI - The role of the pharmacist in humanitarian aid in Bosnia-Herzegovina: the experience of Pharmaciens Sans Frontieres. AB - BACKGROUND: Founded in 1985, Pharmaciens Sans Frontieres (PSF) is a nongovemmental organization of pharmacists involved in humanitarian aid. PSF relied on approximately 100 expatriates in 1998, which included 50 pharmacists distributed throughout 24 missions (i.e., 14 emergency, 7 development, 3 assessment). It is necessary to add 200-250 local staff to this group. OBJECTIVE: To describe PSF's mission in Bosnia-Herzegovina from 1992 to 1999 and to define the pharmacist's impact in the supply of medicines and the development of pharmaceutical care and services. RESULTS: In April 1992, at the beginning of Sarajevo's siege, PSF sent a small team of three volunteer pharmacists to Bosnia Herzegovina. The objective of the emergency phase (1992-1995) was to set up a massive supply program of essential medicines and medical and biologic materials and to implement a distribution system based on existing health centers. The signing of the Dayton peace agreement and a progressive return to peace and stability marked the beginning of the postemergency phase (1995-1997). This phase pursued previous objectives of establishing a distribution network and added the implementation of logistic centers. PSF widened its involvement to medical laboratory analysis, production of medicines, disposal of expired medications sent during the conflict, and the implementation of a national center for quality control. Currently, the development phase's (1998-1999) objective is to provide adequate support for the reorganization of pharmaceutical care and services by establishing pharmacy work groups and developing and maintaining good relationships with the international community and Bosnia-Herzegovina pharmacists. CONCLUSIONS: Humanitarian aid is essential in major conflicts, as seen in the case of Bosnia-Herzegovina. Although it is difficult to evaluate the impact of the distribution network implemented by PSF, it allowed for a better provisioning of medications to the general population. PSF played an important role in such cases. In fact, PSF provides its pharmaceutical expertise to these embattled areas not only by offering financial support to the logistics or supplying of medications, but by offering professional support to the organization/reorganization of the pharmaceutical practice. PMID- 10669195 TI - Glimepiride-induced thrombocytopenic purpura. PMID- 10669196 TI - Effect of azathioprine on the anticoagulant activity of warfarin. PMID- 10669197 TI - Carbamazepine and/or fluvoxamine drug interaction with risperidone in a patient on multiple psychotropic medications. PMID- 10669198 TI - PKBase: a population approach-oriented database. PMID- 10669199 TI - ADEM: literature review and case report of acute psychosis presentation. AB - Acute disseminated encephalomyelitis is a monophasic, immune-mediated disorder that produces multifocal demyelinating lesions within the central nervous system. It is characterized clinically by the acute onset of neurologic abnormalities, including varying degrees of mental state changes ranging from drowsiness to coma. It is unusual for the illness to present as an isolated acute psychosis. The case of a 14-year-old female with biopsy-confirmed acute disseminated encephalomyelitis, who was initially diagnosed with an acute psychiatric disorder, is presented, and published reports on this unusual manifestation are reviewed. A Medline database search was performed from 1965 to 1999, using the terms acute disseminated encephalomyelitis, postvaccinal encephalomyelitis, postinfectious encephalomyelitis, and measles encephalomyelitis, combined with the terms psychosis, psychiatric disorder, and behavioral disorder. Selected cross-referenced reports were also reviewed. Nine patients were identified who presented with acute psychosis. We conclude that, although rare, acute disseminated encephalomyelitis can present as an acute psychosis. This immune mediated condition should be included in the differential diagnosis of neurologic disorders presenting as a psychiatric illness. PMID- 10669200 TI - Intranasal midazolam as a treatment of autonomic crisis in patients with familial dysautonomia. AB - To evaluate the efficacy and safety of intranasal midazolam in the treatment of autonomic crises in children with familial dysautonomia, intranasal midazolam was administered at the hospital to six patients during nine episodes of autonomic crisis. Treatment was successful in seven of nine episodes of autonomic crisis in five of six patients, with a mean interval to response of 9.25 +/- 1.25 minutes. The parents of five patients in whom the treatment was successful were instructed to use midazolam at home. At home, 16 additional episodes were treated by the parents, with successful control achieved in 14 (87%). The mean interval to response was 12.8 +/- 2 minutes. No significant side effects were observed at the hospital or at home after intranasal administration of midazolam. Midazolam, given intranasally, is effective and safe in the management of autonomic crises in patients with familial dysautonomia, either in the hospital or at home by the parents after appropriate instruction. PMID- 10669201 TI - Generalized spike-and-wave patterns in children: clinical correlates. AB - All electroencephalograms performed in our institution between 1980 and 1990 were reviewed. The clinical characteristics of children with epilepsy and generalized spike-and-wave (SW) patterns were analyzed. The SW patterns were classified according to their frequency. Electroencephalograms of 154 children with epilepsy revealed SW patterns. Absence seizures were the most common first seizure, but partial seizures were frequent. More than 40% had several types of seizures. Sixty percent of the epileptic syndromes were generalized, but almost 25% were partial. The typical SW pattern was associated with absence seizures, a normal examination and computed tomographic scan, idiopathic generalized epilepsies, monotherapy, freedom from seizures, and lack of recurrence. The slow SW pattern was associated with West syndrome; a younger age at seizure onset; atonic, myoclonic, tonic, and partial simple seizures; an abnormal examination and computed tomographic scan; cryptogenic or symptomatic generalized epilepsy or symptomatic partial epilepsy; polytherapy; and poor seizure control. The fast SW pattern was associated with secondary generalized, partial, tonic-clonic, and complex partial seizures; a normal computed tomographic scan; cryptogenic partial epilepsy; isolated seizures; and seizure recurrence. Epilepsy with a typical SW pattern should be considered benign, epilepsy with a slow SW pattern malignant, and epilepsy with a fast SW pattern treacherous. PMID- 10669202 TI - Clinical and molecular studies in three Portuguese mtDNA T8993G families. AB - The T8993G mutation in the mitochondrial DNA adenosine triphosphatase 6 gene represents an important cause of maternally inherited Leigh's syndrome. Reported are the clinical findings and mutational loads in three Portuguese T8993G pedigrees. Polymerase chain reaction-restriction fragment length polymorphism analyses demonstrated the T8993G mutation in a high percentage of tissues from all patients (97% +/- 2.3%), but it was less abundant in the blood from 14 maternal relatives. The disease progressed severely in the probands but did not have the fatal course reported by others. To test whether this prolonged course was related to the presence of a specific, disease-associated haplogroup the origin of the mutational event in Portugal was traced. Haplotype investigation revealed an independent occurrence of the mutation in the three probands. These analyses represent the first molecular characterization of Portuguese patients with Leigh's syndrome. PMID- 10669203 TI - Sleep apnea treatment improves seizure control in children with neurodevelopmental disorders. AB - Seizure disorder and sleep apnea are common chronic disorders in children, but the relationship between sleep apnea and seizure control has not been studied in the pediatric population. This retrospective review included nine children with neurodevelopmental disorders who had well-documented sleep apneic episodes and seizure disorders. Seizure frequency was reduced in five patients (56%) in the first 12 months after sleep apnea treatment without changes in their antiepileptic medications. Sleep apnea can be one of the seizure precipitants in children with epilepsy. This study indicates the importance of identifying sleep apnea when treating children with intractable epilepsy, particularly in those who are at high risk. PMID- 10669204 TI - High-dose intravenous immunoglobulin therapy in juvenile myasthenia gravis. AB - Autoimmune neurologic disease management has been significantly modified by the use of high-dose intravenous immunoglobulin (HDIVIG) during the past 15 years. Venous access, readily available IgG (until recently), and the relative lack of serious identifiable complications have prompted its use in myasthenia gravis. In adults, its effectiveness has been inconsistent, with variable acetylcholine receptor (AChR) antibody responses. Ten children were evaluated for clinical responses to, and complications of, HDIVIG. Weekly anti-AChR antibody titers in three patients were obtained. The HDIVIG dosage was 2 gm/kg body weight, infused at variable rates of 2 gm/kg for 1 day, 0.66 gm/kg daily for 3 days, and 0.5 g/kg daily for 4 days; in one patient the total dose was 0.8 gm/kg to correct to the ideal body weight. All children but one tolerated HDIVIG without complications. Eight patients exhibited definite improvement in functional strength after HDIVIG, but a decreasing response to HDIVIG was evident after multiple monthly treatments, warranting the additional use of corticosteroids in two patients. A decrease in anti-AChR antibody levels was observed in the three patients tested, but this decrease was constant in one patient. No correlation was observed between clinical response and antibody titers. HDIVIG is safe and effective in most patients for short-term management of juvenile myasthenia gravis, in myasthenic crises, and in preparing patients for surgery but appears to be of limited long-term benefit. PMID- 10669205 TI - Clinical and cerebral FDG PET scan in a patient with Krabbe's disease. AB - A 2-year, 6-month-old Saudi male with infantile Krabbe's disease was studied with fluorine-18-labeled-2-fluoro-2-deoxyglucose positron emission tomography (FDG PET) scan. The patient presented with a gradual loss of developmental milestones, irritability, and crying. At the advanced stage of the disease, he developed tonic-clonic seizures and became a microcephalic, extremely irritable, blind, spastic quadriplegic child, with no deep tendon reflexes. Laboratory studies revealed normal blood chemistry, muscle enzymes, very long chain fatty acids, and acylcarnitines. No abnormal urinary organic acids were detected. The cerebrospinal fluid protein concentration was increased. Magnetic resonance imaging of the brain revealed mild brain atrophy and white matter disease mainly in the centrum semiovale. Electroretinography was normal; however, electroencephalography and visual-evoked potentials were abnormal. Peripheral nerve conduction studies documented a demyelinating neuropathic process. The FDG PET study of the brain demonstrated a marked decrease in the metabolism of the left cerebral cortex and no uptake in the caudate heads. Normal glucose uptake was observed in the thalami, lentiform nuclei, and cerebellum. The patient did not present for subsequent clinic visits and is presumed dead. PMID- 10669206 TI - Potential hepatotoxicity of lamotrigine. AB - Lamotrigine is a new antiepileptic drug that is effective for a broad range of seizures in adults and children. Three children with seizures of different causes who were treated with lamotrigine and developed reversible hepatotoxicity are reported. In one child, this therapy led to relatively severe hepatic failure that required and responded to aggressive therapy. Unlike most of the previously reported six patients with similar severe hepatic involvement, this patient's liver function and blood hepatic enzymes became normal. All three patients were on multiple drugs, and two were in epilepsia partialis continua secondary to encephalitis. Two of the patients had relatively rapid medication titration schedules. The close time relationship between the initiation of the lamotrigine therapy and the reversal of the liver abnormalities with lamotrigine discontinuation argues against a cause other than the lamotrigine; however, because of the complexity of the reported cases, the causality remains an assumption. Review of the literature revealed six other previously reported patients (five adults and one child) who had hepatotoxicity during lamotrigine therapy, with or without associated multisystem failure, and similar patient profiles. Lamotrigine is generally a safe and effective medication; however, it should be used with caution in patients on polytherapy and in those with complicated acute systemic and central nervous system conditions, such as fever, status epilepticus, epilepsia partialis, and encephalitis. PMID- 10669207 TI - Mitochondrial complex I deficiency in a female with multiplex arthrogryposis congenita. AB - A 10-year-old female with arthrogryposis multiplex congenita is presented. Clinical, neurophysiologic, and histologic findings suggested a mild myopathy. The analysis of enzymatic activity in the homogenate and of mitochondrial function in saponin-permeabilized fibers from the muscle biopsy revealed an approximately twofold-decreased specific activity of the NADH:CoQ oxidoreductase (complex I of the mitochondrial respiratory chain) that was compensated for by an increased number of mitochondria. The complex I deficiency was also detected in cultivated skin fibroblasts of the patient. The observed defect of mitochondrial oxidative phosphorylation in arthrogryposis multiplex congenita may be of pathogenetic relevance. PMID- 10669208 TI - Neuroblastoma associated with seizures and arrested development. AB - Two unrelated cases of childhood peripheral neuroblastoma associated with infantile seizures and developmental problems (but without opsoclonus-myoclonus) are presented. The considerable body of evidence supporting the view that the opsoclonus-myoclonus syndrome associated with neuroblastoma has an immunologic basis is reviewed. Patients with neuroblastoma and opsoclonus-myoclonus syndrome commonly have subsequent developmental problems and, rarely, may have seizures. The authors postulate that the seizures and developmental problems in their two patients may result from an immunologic mechanism similar to that suggested for the opsoclonus-myoclonus syndrome of neuroblastoma. The only laboratory evidence to support an immunologic mechanism in these two patients was the presence of raised cerebrospinal fluid immunoglobulins in Patient 2. Specific antineuronal antibody tests in Patient 2 were negative. It is therefore possible that the association reported in these two unrelated cases is coincidental. However, reasonably extensive investigations did not uncover an alternative etiology for the presence of the seizures and developmental delay. PMID- 10669209 TI - Aggressive behavior in patients with Sotos syndrome. AB - Sotos syndrome is characterized by peculiar facies, prenatal and postnatal overgrowth, and developmental delay. The course of six patients with psychiatric, neurologic, and magnetic resonance imaging evaluations is reported. Three (50%) of the six patients were observed to have a tendency toward aggressiveness, including one who had pyromania. PMID- 10669210 TI - Gabapentin treatment in a child with delayed-onset hemichorea/hemiballismus. AB - A 13-year, 6-month-old female was evaluated for subacute onset of left-sided hemichorea/hemiballismus, with an old, right parietal, cortical, and subcortical stroke as the presumed cause. Treatment with gabapentin was initiated, with good results at 6-month follow-up. Discussion of the differential diagnosis and evaluation of delayed-onset movement disorders in children and the mechanism of action of gabapentin is included. PMID- 10669211 TI - Congenital intracranial teratoma. AB - Congenital intracranial teratoma is a rare disease. A fetus with a congenital intracranial teratoma presenting with a disproportionately enlarged head at 27 weeks gestation is presented. Prenatal ultrasonography and fetal magnetic resonance imaging demonstrate a huge, heterogenous intracranial mass in the left supratentorial region, with the left cerebral hemisphere being compressed and flattened. The infant died of respiratory failure within 24 hours of birth at 28 weeks gestation. On postmortem examination the histologic report revealed an immature teratoma. Fetal MRI is helpful in the prenatal diagnosis and evaluation of intracranial tumor. PMID- 10669212 TI - Plantar grasp reflex in high-risk infants during the first year of life. AB - For most primitive reflexes, retention of the reflex beyond the period when it should no longer be elicited suggests a pathologic process within the central nervous system. However, for certain primitive reflexes, such as the plantar grasp reflex, a negative response within the first months of life is suggestive of a neurologic abnormality. From the results of one prospective and one retrospective study, it is clearly indicated that the absence of the plantar grasp reflex from 3 months of age and on correlates with the development of spastic cerebral palsy. The specific combination of presence or absence of specific primitive reflexes, postural reactions, or both may accurately predict a specific type of cerebral palsy or neurodevelopmental abnormality. PMID- 10669213 TI - Spine imaging. PMID- 10669215 TI - Interventional neuroradiology. PMID- 10669214 TI - Brain imaging. PMID- 10669216 TI - Head and neck imaging. PMID- 10669217 TI - Pediatric neuroradiology. PMID- 10669218 TI - The future of neurologic MR imaging. PMID- 10669219 TI - Implications of a reliable method for quantifying brain injury associated with repair of carotid artery stenosis. PMID- 10669220 TI - Staining for apoptosis: now neuropathologists can "see" leukoaraiosis. PMID- 10669221 TI - The residents did not miss many? Are you kidding? PMID- 10669222 TI - Guidelines for diagnostic neuroangiography: a model to emulate from a neuroradiologist's perspective. PMID- 10669223 TI - Toward an understanding of syringomyelia: MR imaging of CSF flow and neuraxis motion. PMID- 10669224 TI - Ischemia after carotid endarterectomy: comparison between transcranial Doppler sonography and diffusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: Hyperintense signals on diffusion-weighted MR images (DWIs) are believed to correspond accurately with cerebral ischemic events. Intraoperative transcranial Doppler sonography (TCD) can reveal hemodynamic and embolic events during carotid endarterectomy (CEA). Our purpose was to determine whether the occurrence of hyperintense signals on postoperative DWIs corresponds to intraoperative embolic or hemodynamic events. METHODS: Seventy-seven CEAs were monitored intraoperatively with TCD to record blood flow velocity changes after cross clamping to ascertain the presence of adequate collateral flow and to record microembolic signals. DWI was used to classify the hemisphere ipsilateral to the CEA by type: 0, no lesions (n = 51); I, cortical lesions only (n = 2); II, subcortical white matter lesions only (n = 6); III, mixed type with cortical and subcortical lesions (n = 11); IV, large territorial infarcts (n = 6); and V, other types of lesions (n = 1). RESULTS: Neither the five clinical events (one transient ischemic attack, two minor strokes, and two major strokes) nor any DWI type (I-V) showed a relationship to blood velocity decreases after cross clamping or, in patients who were selectively shunted, to total ischemic time necessary for shunt insertion and removal. Total microembolic signal count was significantly higher in the five CEAs with clinical events than in those without. It was also higher on the DWIs showing a hyperintense lesion as compared with DWIs showing no lesion. CONCLUSION: Apart from lesions corresponding to clinical deficits, CEA is associated with a substantial number of small areas of brain tissue at risk for irreversible ischemia. The main cause of intraoperative stroke seems to be embolism, suggesting that microembolic signals in CEA are highly relevant events for brain tissue. PMID- 10669225 TI - Incidence of postangiographic abnormalities revealed by diffusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: Occasionally we have observed anecdotal cases of asymptomatic hyperintensities on diffusion-weighted MR (DW-MR) examinations of the brain of patients who previously underwent routine cerebral angiography. These observations, as well as MR imaging and transcranial Doppler data in the literature suggesting a high rate of procedure-associated emboli, raise concern regarding the underdiagnosis of asymptomatic focal infarction associated with cerebral angiography. In order to determine whether asymptomatic diffusion abnormalities are frequently associated with procedures, we prospectively obtained DW-MR images before and after routine cerebral angiography. METHODS: Twenty consecutive patients, who met protocol criteria and received a routine three- or four-vessel diagnostic cerebral angiogram at our institution, were evaluated. Using a Bayesian estimate to establish an upper bound for the incidence of an event with zero occurrences in a study sample, the study group size was selected to exclude a 10% incidence of abnormalities revealed by DW-MR imaging of patients who underwent previous cerebral angiography. Two neuroradiologists evaluated imaging studies. RESULTS: Neither clinical signs nor abnormalities on DW-MR images were found, which suggested no infarction after angiography in our patient sample. Based on this data, an upper bound of 9% (95% confidence) is predicted for the appearance of abnormalities revealed by DW-MR imaging after cerebral angiography. CONCLUSION: Cerebral angiography is associated with an incidence of asymptomatic cerebral infarction of no more than 9%. It well may be substantially lower than this estimate; a more accurate evaluation of the true incidence would require a significantly larger study population. This test provides a convenient noninvasive means of assessing procedure-related cerebral infarction, such as that which occurs after carotid endarterectomy or vascular angioplasty and stenting. PMID- 10669226 TI - Histopathologic correlates of temporal diffusion changes in a rat model of cerebral hypoxia/ischemia. AB - BACKGROUND AND PURPOSE: Although diffusion-weighted MR imaging is a powerful tool for evaluating brain ischemia, histopathologic correlates of temporal diffusion changes in cerebral hypoxia/ischemia have not been extensively examined. Diffusion-weighted MR imaging was used to evaluate the relationship between the time course of apparent diffusion coefficient (ADC) changes and the histopathologic findings in cerebral hypoxia/ischemia. METHODS: Thirty 3-week-old rats were subjected to either a 15-, 30-, or 60-minute hypoxic/ischemic insult (unilateral common carotid artery ligation and exposure to 8% oxygen), during and after which diffusion- and T2-weighted MR imaging was performed. Each animal was killed 48 hours or 6 hours after the insult, and fixed sections of the parietal cortex were examined by light microscopy. Ten other (control) rats were subjected to only unilateral common carotid artery ligation or hypoxia. RESULTS: The experimental rats showed three patterns of ADC change, depending on the duration of the hypoxic/ischemic insult: transient (15-minute), biphasic (15-, 30-, or 60 minute), and persistent (60-minute) ADC reduction patterns. The transient ADC reduction pattern (reduction during the insult and recovery after resuscitation) was associated with selective neuronal death. The biphasic and persistent ADC reduction patterns (transient recovery and no recovery after resuscitation, respectively) were associated with cerebral infarction. CONCLUSION: Different temporal patterns of ADC change are associated with different histopathologic findings. Although the clinical manifestations of these different histopathologic presentations are not yet defined, this study indicates that sequential diffusion studies are a potentially powerful tool in the evaluation of hypoxic/ischemic brain injury. PMID- 10669227 TI - Diffusion- and perfusion-weighted MR imaging of dural sinus thrombosis. AB - A patient with dural sinus thrombosis had progressively worsening symptoms and signs that resolved after intradural thrombolysis. Intradural sinus pressures were 54 mm Hg. Echo-planar MR imaging revealed complex abnormalities of diffusion and widespread delay in mean transit time that improved immediately after thrombolysis. This case suggests that diffusion- and perfusion-weighted imaging can provide valuable information noninvasively to help triage patients with dural sinus thrombosis between conservative and aggressive management. PMID- 10669228 TI - Cerebral MR venography: normal anatomy and potential diagnostic pitfalls. AB - BACKGROUND AND PURPOSE: MR venography is often used to examine the intracranial venous system, particularly in the evaluation of dural sinus thrombosis. The purpose of this study was to evaluate the use of MR venography in the depiction of the normal intracranial venous anatomy and its variants, to assess its potential pitfalls in the diagnosis of dural venous sinus thrombosis, and to compare the findings with those of conventional catheter angiography. METHODS: Cerebral MR venograms obtained in 100 persons with normal MR imaging studies were reviewed to determine the presence or absence of the dural sinuses and major intracranial veins. RESULTS: Systematic review of the 100 cases revealed transverse sinus flow gaps in 31% of the cases, with 90% of these occurring in the nondominant transverse sinus and 10% in the codominant transverse sinuses. No flow gaps occurred in the dominant transverse sinuses. The superior sagittal and straight sinuses were seen in every venogram; the occipital sinus was seen in only 10%. The vein of Galen and internal cerebral veins were also seen in every case; the basal veins of Rosenthal were present in 91%. CONCLUSIONS: Transverse sinus flow gaps can be observed in as many as 31% of patients with normal MR imaging findings; these gaps should not be mistaken for dural sinus thrombosis. PMID- 10669229 TI - Apoptosis in leukoaraiosis. AB - We report a case of leukoaraiosis that was studied for apoptosis. In the neuropil, the number of cells that showed DNA fragmentation was 2.5 times as great in the area of leukoaraiosis as in the adjacent white matter (P = .004) and 25 times as great as in the nearby cortex (P < .001). Our findings suggest that apoptosis, predominantly of oligodendrocytes, is involved in the pathogenesis of leukoaraiosis. Within the area of leukoaraiosis, we also found numerous small veins that were partially occluded by severe collagenous thickening of the vessel walls. This collagenosis may have contributed to or resulted from chronic ischemia in that area. PMID- 10669230 TI - Proton MR spectroscopy and preoperative diagnostic accuracy: an evaluation of intracranial mass lesions characterized by stereotactic biopsy findings. AB - BACKGROUND AND PURPOSE: MR imaging has made it easier to distinguish among the different types of intracranial mass lesions. Nevertheless, it is sometimes impossible to base a diagnosis solely on clinical and neuroradiologic findings, and, in these cases, biopsy must be performed. The purpose of this study was to evaluate the hypothesis that proton MR spectroscopy is able to improve preoperative diagnostic accuracy in cases of intracranial tumors and may therefore obviate stereotactic biopsy. METHODS: Twenty-six patients with intracranial tumors underwent MR imaging, proton MR spectroscopy, and stereotactic biopsy. MR spectroscopic findings were evaluated for the distribution pattern of pathologic spectra (NAA/Cho ratio < 1) across the lesion and neighboring tissue, for signal ratios in different tumor types, and for their potential to improve preoperative diagnostic accuracy. RESULTS: Gliomas and lymphomas showed pathologic spectra outside the area of contrast enhancement while four nonastrocytic circumscribed tumors (meningioma, pineocytoma, metastasis, and germinoma) showed no pathologic spectra outside the region of enhancement. No significant correlation was found between different tumor types and signal ratios. MR spectroscopy improved diagnostic accuracy by differentiating infiltrative from circumscribed tumors; however, diagnostic accuracy was not improved in terms of differentiating the types of infiltrative or circumscribed lesions. CONCLUSION: MR spectroscopy can improve diagnostic accuracy by differentiating circumscribed brain lesions from histologically infiltrating processes, which may be difficult or impossible solely on the basis of clinical or neuroradiologic findings. PMID- 10669231 TI - Single- and multiple-event paradigms for identification of motor cortex activation. AB - BACKGROUND AND PURPOSE: The "single-event" technique has been used as an alternative to the "block-trial" method to detect activation that may be accompanied by head motion. The purpose of this study was to compare the two methods for measuring activation in the sensorimotor cortex secondary to motor tasks. METHODS: Functional MR imaging data were acquired from six participants as they performed tasks with their fingers, tongues, and toes in a block-trial and a single-event paradigm. For the block trial, the participant was instructed to perform the task when cued at a rapid self-timed rate for 15 seconds, alternating with 15 seconds of rest. Five periods of task performance and six rest periods were included in one acquisition. For the single-event method, the participant performed the task a single time every 15 seconds when cued by the investigator, for a total of 21 times. Using conventional parcellation methods, activation was detected by a cross-correlation technique and was classified as occurring in the sensorimotor cortex, supplementary motor area (SMA), or as nonspecific. Differences between the two acquisition paradigms were tested using the standard t test at a significance level of P < .05. RESULTS: Activation was identified by both the block-trial and the single-event methods for the finger task, for the tongue task, and inconsistently for the toe task. More motion artifact occurred in conjunction with the toe and tongue tasks than with the finger tasks. On average, more activated pixels were identified by the single-event method than by the block-trial method. For these motor tasks, however, a larger percentage of pixels detected by the block-trial method than by the single-event method were specific for the sensorimotor cortex or SMA as sites of activation. CONCLUSION: For the tongue and the toe movement tasks, which may produce some head motion artifacts, the single-event paradigm provides a useful alternative to the block trial method for identifying the sensorimotor cortex or SMA. It does not achieve a greater percentage of activation within primary motor areas. For the finger movement task, which does not usually produce head motion artifacts, the block trial method generally produced a greater percentage of activated pixels in the sensorimotor cortex or SMA than did the single-event method. PMID- 10669232 TI - Brain fluorine-18 fluorodeoxyglucose imaging with dual-head coincidence gamma camera: comparison with dedicated ring-detector positron emission tomography. AB - BACKGROUND AND PURPOSE: Dual-head coincidence gamma camera (DHC) imaging has been proposed as an alternative to dedicated ring-detector positron emission tomography (dr-PET) for clinical fluorodeoxyglucose (FDG) studies. The purpose of this investigation was to assess the quality of DHC images in FDG studies of the human brain. METHODS: Seven healthy volunteers and 12 patients with various cerebral disorders underwent consecutive brain dr-PET and DHC with FDG. All sets of images were compared semiquantitatively using regions of interest. RESULTS: Cortical count ratios to the cerebellum on DHC and dr-PET images did not differ significantly among the volunteers, except in the superior frontal cortex and thalamus. In all studies including those of cerebral disorders, the mean cortical to-cerebellar ratios of DHC and dr-PET images correlated closely. CONCLUSION: FDG imaging with DHC delineated the metabolic distribution of glucose in the brain as well as dr-PET did, except in the superior frontal cortex and thalamus. Therefore, DHC may be a dedicated cost-effective means of detecting metabolic abnormalities in the brain. PMID- 10669233 TI - Visualization of intravenously administered contrast material in the CSF on fluid attenuated inversion-recovery MR images: an in vitro and animal-model investigation. AB - BACKGROUND AND PURPOSE: The FLAIR (fluid-attenuated inversion-recovery) pulse sequence has been shown to be sensitive to abnormalities of the subarachnoid space. Our clinical experience led us to investigate whether intravenously injected contrast material can affect the appearance of the subarachnoid space on FLAIR MR images. METHODS: After noting unexplained high signal in the subarachnoid space on FLAIR images in a patient, we studied two dogs with sequential FLAIR MR imaging after i.v. administration of contrast material. A third dog was studied with a 6-hour delayed FLAIR sequence after triple-dose (0.3 mmol/kg) i.v. contrast administration. CSF was obtained from two animals for measurement of gadolinium concentration. A phantom was developed to determine the lowest concentration at which the effects of gadolinium were evident on FLAIR images in vitro. RESULTS: In all three animals, the appearance of the CSF in the ventricles or subarachnoid space was modified after administration of i.v. contrast. This was most evident on delayed images. The CSF samples showed a gadolinium concentration of 0.007 mmol/L in the dog who received the 0.1 mmol/kg dose and 0.02 mmol/L in the dog who received a triple dose. In our in vitro phantom experiments, gadolinium effects were evident on FLAIR images at a concentration four times lower than those on T1-weighted images. CONCLUSION: I.v. contrast material can cross into the CSF in sufficient concentration to alter the appearance of the subarachnoid space on FLAIR images in normal dogs. Although we encountered two patients with CNS disease in whom enhancement of the CSF was seen on postcontrast FLAIR images, additional investigation is needed in humans to determine whether enhancement may occur at triple dose in healthy subjects. PMID- 10669234 TI - Gender effects on age-related changes in brain structure. AB - BACKGROUND AND PURPOSE: Previous reports have suggested that brain atrophy is associated with aging and that there are gender differences in brain atrophy with aging. These reports, however, neither exclude silent brain lesions in "healthy subjects" nor divide the brain into subregions. The aim of this study is to clarify the effect of gender on age-related changes in brain subregions by MR imaging. METHODS: A computer-assisted system was used to calculate the brain matter area index (BMAI) of various regions of the brain from MR imaging of 331 subjects without brain lesions. RESULTS: There was significantly more brain atrophy with aging in the posterior parts of the right frontal lobe in male subjects than there was in female subjects. Age-related atrophy in the middle part of the right temporal lobe, the left basal ganglia, the parietal lobe, and the cerebellum also was found in male subjects, but not in female subjects. In the temporal lobe, thalamus, parieto-occipital lobe, and cerebellum, brain volume in the left hemisphere is significantly smaller than in the right hemisphere; sex and age did not affect the hemisphere differences of brain volume in these regions. CONCLUSION: The effect of gender on brain atrophy with aging varied in different subregions of the brain. There was more brain atrophy with aging in male subjects than in female subjects. PMID- 10669235 TI - Serial CT and MR imaging of carmustine wafers. AB - BACKGROUND AND PURPOSE: A new option in the treatment of recurrent malignant glioma is surgical placement of chemotherapy-laden biodegradable wafers. We describe the CT and MR appearance of chemotherapy wafers in patients after surgery for recurrent malignant glioma METHODS: Eighteen patients had carmustine (BCNU) wafers implanted during reoperation for malignant glioma; three patients had empty, placebo wafers placed. The 21 patients had a total of 22 CT and 57 MR imaging studies. Repeat CT studies were conducted for up to 6 months, the MR studies for up to 1 year. Examinations were evaluated for attenuation on CT scans, signal abnormalities on MR images, and changing appearance during the follow-up period. Enhancement characteristics were also assessed. RESULTS: On CT scans, 13 of 16 acute (<7 days) cases showed linear high-attenuation wafers, with three showing low attenuation. On MR images, all T1 and T2 studies performed in the acute stage showed decreased signal of the wafers. Eight of 15 studies showed a transient increase in T1 only at about 2 months. Wafers decreased in conspicuity on both CT and MR studies after 2 months. The wafers did not enhance. One postoperative tumor showed a transient increase in edema and increased enhancement at 5 weeks. The presence or absence of BCNU within the wafers did not change their appearance. CONCLUSION: BCNU wafers have a characteristic appearance: in the first 7 days after implantation they are linear, usually of increased attenuation on CT scans, and always show decreased signal on MR images; they do not enhance, and become less conspicuous after 2 months. PMID- 10669236 TI - Clinical consequences of misinterpretations of neuroradiologic CT scans by on call radiology residents. AB - BACKGROUND AND PURPOSE: Studies have looked at the accuracy of radiologic interpretations by radiology residents as compared with staff radiologists with regard to emergency room plain films, emergency room body CT scans, and trauma head CT scans; however, to our knowledge, no study has evaluated on-call resident interpretations of all types of neuroradiologic CT scans. Both as a part of our departmental quality control program and to address concerns of clinical services about misinterpretation of neuroradiologic CT scans by on-call radiology residents, we evaluated the frequency of incorrect preliminary interpretations of neuroradiologic CT scans by on-call radiology residents and the effect of such misinterpretations on clinical management and patient outcome. METHODS: As determined by the staff neuroradiologist the next day, all potentially clinically significant changes to preliminary reports of emergency neuroradiologic CT scans rendered by on-call radiology residents were recorded over a 9-month period. A panel of neuroradiologists reviewed and graded all the changed cases by consensus. An emergency department staff physician reviewed medical records of all submitted cases to determine clinical consequences of the misinterpretations. RESULTS: Significant misinterpretations were made in 21 (0.9%) of 2388 cases during the study period. There was a significant change in patient management in 12 of the cases, with a potentially serious change in patient outcome in two cases (0.08%). CONCLUSION: On-call radiology residents have a low rate of significant misinterpretations of neuroradiologic CT scans, and the potential to affect patient outcome is rare. PMID- 10669237 TI - MR imaging features of clear-cell meningioma with diffuse leptomeningeal seeding. AB - Clear-cell meningioma is a rare disease entity showing a more aggressive nature, clinically, than those of other subtypes of meningioma. It occurs in younger persons and commonly in the spinal canal. The recurrence rate has been reported to be as high as 60%. We present a case of clear-cell meningioma in a 17-year-old man in whom initial MR imaging showed localized leptomeningeal enhancement that had progressed into the entire subarachnoid space after surgical resection of the primary tumor. PMID- 10669238 TI - Reproducibility of magnetization transfer ratio histogram-derived measures of the brain in healthy volunteers. AB - Using two MR scanners, we evaluated the intraobserver, interobserver, image reimage, and interimager variabilities in the assessment of magnetization transfer ratio (MTR) histograms obtained monthly on four occasions from five healthy volunteers. With multiple observers, the mean coefficients of variations ranged from 2.2% to 8.2% for "pure" image-reimage variability, from 1.2% to 4.9% for interobserver variability, and from 2.1% to 4.9% for image-reimage variability. The mean intraobserver coefficients of variations were always lower than 1%. The mean coefficients of variations ranged from 10.2% to 14.6% for pure interimager variability and from 8.6% to 14.3% for interimager variability with multiple observers. Interimager variability accounted for 96.0% of the overall variability of average MTR, for 96.7% of peak location, and for 41.1% of the peak height. The use of different MR scanners is the main source of variability when obtaining MTR histograms. PMID- 10669239 TI - Coil occlusion of the parent artery for treatment of symptomatic peripheral intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Peripheral intracranial aneurysms can be difficult to treat with traditional surgical or embolization techniques that spare the parent vessel. We report the results of our use of coil occlusion of the parent vessel for the treatment of nine peripheral intracranial aneurysms. METHODS: During approximately a 4-year period, nine patients (six men and three women, 27 to 68 years old; average age, 42 years) presented to our institution with peripheral intracranial aneurysms. The aneurysms were located on branches of the right posterior inferior cerebellar artery (n = 2), the right superior cerebellar artery (n = 1), the right anterior inferior cerebellar artery (n = 1), the right posterior cerebral artery (n = 3), the left middle cerebral artery (n = 1), and the left anterior cerebral artery (n = 1). Parent vessel occlusion was performed using microcoils after test injection with amobarbital (Amytal) in eight of the nine cases (one patient was comatose and could not be tested before occlusion). RESULTS: Angiography immediately after the procedure showed aneurysmal occlusion in every patient. Follow-up arteriography, performed in six patients 2 to 12 months after treatment, documented continued aneurysmal occlusion in every case. Three patients exhibited mild, nondisabling neurologic deficits after coil placement; the rest had no new deficits, although one patient was severely disabled from the initial hemorrhage and one patient died of an unrelated cause. CONCLUSION: Our results lend support to the use of parent vessel occlusion for peripheral aneurysms that are difficult to treat surgically or that are not amenable to intra-aneurysmal coil placement. PMID- 10669240 TI - Selective infusion of urokinase and thrombectomy in the treatment of acute cerebral sinus thrombosis. AB - Acute cerebral sinus thrombosis caused a patient to decompensate rapidly and required immediate relief of her venous thrombosis as a life-saving procedure. The thrombus was laced with urokinase and removed from the sinuses with thrombectomy catheters. This reinstituted flow and she recovered full neurologic function within 4 hours. PMID- 10669241 TI - Quality improvement guidelines for adult diagnostic neuroangiography. Cooperative study between the ASNR, ASITN, and the SCVIR. American Society of Neuroradiology. American Society of Interventional and Therapeutic Neuroradiology. Society of Cardiovascular and Interventional Radiology. PMID- 10669242 TI - Phase-contrast MR imaging of the cervical CSF and spinal cord: volumetric motion analysis in patients with Chiari I malformation. AB - BACKGROUND AND PURPOSE: Most previous MR studies of the dynamics of Chiari I malformation have been confined to sagittal images and operator-dependent measurement points in the midline. To obtain a deeper insight into the pathophysiology of the Chiari I malformation, we performed a prospective study using axial slices at the level of C2 to analyze volumetric motion data of the spinal cord and CSF over the whole cross-sectional area. METHODS: Eighteen patients with Chiari I malformation and 18 healthy control subjects underwent cardiac-gated phase-contrast imaging. Cross-sectional area measurements and volumetric flow/motion data calculations were made for the following compartments: the entire intradural space, the spinal cord, and the anterior and posterior subarachnoid space. RESULTS: The most striking feature was an increased early systolic caudal and diastolic cranial motion of the spinal cord in the patients. CSF pulsations in the anterior subarachnoid space were unchanged at systole but showed an impaired diastolic upward flow. In the posterior compartment, the CSF systole was slightly shortened, with an impairment of diastolic upward flow. Fourteen of the 18 patients had associated syringeal cavities. This subgroup showed an increased systolic downward displacement of the cord as compared with patients without a syrinx. CONCLUSION: Obstruction of the foramen magnum in patients with Chiari I malformation causes an abrupt systolic downward displacement of the spinal cord and impairs the recoil of CSF during diastole. PMID- 10669243 TI - Percutaneous vertebroplasty: a special syringe for cement injection. AB - Percutaneous vertebroplasty is an effective treatment for many focal vertebral lesions. Methyl methacrylate is too viscous to be handled without difficulty in the conventional way because injection time is short. The operator is left with little time and must fumble with multiple syringes. We describe a special screw system syringe that decreases the effort needed to inject the cement. In addition, it can standardize the injection pressures and control the injected volume because the threaded plunger affords greater control of injection pressure and volume delivered than does the conventional method. PMID- 10669244 TI - Vascularization of head and neck paragangliomas: comparison of three MR angiographic techniques with digital subtraction angiography. AB - BACKGROUND AND PURPOSE: MR angiography of the head and neck region has been studied widely, but few studies have been performed concerning the efficacy of MR angiography for the identification of the specific vascular supply of the highly vascular head and neck paragangliomas. In this study, we compared three MR angiography techniques with respect to visualization of branch arteries in the neck and identification of tumor feeders in patients with paragangliomas. METHODS: Fourteen patients with 29 paragangliomas were examined at 1.5 T using 3D phase-contrast (PC), 2D time-of-flight (2D TOF), and multi-slab 3D TOF MR angiography. In the first part of the study, two radiologists independently evaluated the visibility of first-, second-, and third-order branch arteries in the neck. In the second part of the study, the number of feeding arteries for every paraganglioma was determined and compared with digital subtraction angiography (DSA), the standard of reference in this study. RESULTS: Three dimensional TOF angiography was superior to the other MR angiography techniques studied (P < .05) for depicting branch arteries of the external carotid artery in the neck, but only first- and second-order vessels were reliably shown. DSA showed a total of 78 feeding arteries in the group of patients with 29 paragangliomas, which was superior to what was revealed by all MR angiography techniques studied. More tumor feeders were identified with 3D TOF and 2D TOF angiography than with 3D PC MR angiography (P < .05), with a sensitivity/specificity of 61%/98%, 54%/95%, and 31%/95%, respectively. Sensitivity was lowest for carotid body tumors. CONCLUSION: Compared with intra arterial DSA, the 3D TOF MR angiography technique was superior to 3D PC and 2D TOF MR angiography for identifying the first- and second-order vessels in the neck. With 3D TOF angiography, more tumor feeders were identified than with the other MR angiography techniques studied. The sensitivity of MR angiography, however, is not high enough to reveal important vascularization. The sensitivity of MR angiography is too low to replace DSA, especially in the presence of carotid body tumors. PMID- 10669245 TI - Imaging and clinical evaluation of isolated atresia of the oval window. AB - BACKGROUND AND PURPOSE: Congenital causes of hearing loss in children commonly are encountered, and imaging aids in diagnosis as well as presurgical evaluation. Atresia of the oval window not associated with atresia of the external auditory canal (EAC) is a rare cause of congenital hearing loss in children. We present the clinical and imaging findings in children with isolated oval-window atresia. METHODS: Atresia of the oval window was defined as the absence of the structure with the presence of a bony plate superimposed between the vestibule and middle ear. The bony plate is within the expected region of the oval window. Using a computerized database, nine patients with isolated oval-window atresia were found. All had been evaluated with high-resolution computed tomography (HRCT) and all had medical records available for review, including audiogram results. Imaging studies were interpreted by the consensus of two pediatric neuroradiologists. RESULTS: Atresia of the oval window was documented in all cases using HRCT criteria. The most common anomalies associated with oval-window atresia were inferomedial malposition of the facial nerve (n = 8), malformed incus (n = 6), and displaced stapes (n = 2). Four patients had symmetric bilateral involvement. Hearing tests were not specific, because conductive, sensorineural, and mixed patterns were found. CONCLUSION: Anomalies of the oval window should be sought in all patients with congenital hearing loss. Associated findings, such as facial nerve aberrancy and ossicular anomalies, are important in both diagnosis and surgical planning. PMID- 10669246 TI - Masticator space abnormalities associated with mandibular osteoradionecrosis: MR and CT findings in five patients. AB - BACKGROUND AND PURPOSE: Imaging of patients with a clinical diagnosis of mandibular osteoradionecrosis (ORN) is often performed to support that clinical suspicion, evaluate the extent of the disease, or exclude coexistent tumor recurrence. The purpose of our study was to describe the clinical, MR imaging, and CT features of five patients with mandibular ORN associated with prominent soft-tissue abnormality in the adjacent masticator muscles. METHODS: The MR and CT examinations of five patients with mandibular ORN associated with soft-tissue abnormalities in the adjacent masticator muscles were reviewed. All patients had received external beam radiotherapy for primary head and neck malignancies, with a total radiation dose range of 60 Gy to 69 Gy in 30 to 38 fractions. RESULTS: CT revealed the typical osseous findings of cortical disruption, trabecular disorganization, and fragmentation in all five patients. Abnormal diffuse enhancement of the adjacent masseter and pterygoid muscles was noted in all patients. Four patients had prominent mass-like thickening of these muscles adjacent to the osseous abnormality. Of the three patients who underwent MR imaging, all showed homogeneous abnormal T1 hypointensity, T2 hyperintensity, and intense enhancement of the bone marrow in the involved mandible. The masticator muscles adjacent to the osseous abnormality also showed abnormal T2 hyperintensity and intense diffuse enhancement on MR images. CONCLUSION: Mandibular ORN can be associated with prominent soft-tissue thickening and enhancement in the adjacent musculature. These changes can appear mass-like and are not related to tumor recurrence or metastatic disease. PMID- 10669247 TI - Age-related expansion and reduction in aeration of the sphenoid sinus: volume assessment by helical CT scanning. AB - BACKGROUND AND PURPOSE: Aeration of the sphenoid sinus expands with the development of the sphenoid bone, but scant detailed volumetric data regarding this process, as it evolves from childhood to old age, exist. Using helical CT scanning, we assessed age-related volumetric changes of the sphenoid sinus. METHODS: We used CT data obtained from 214 patients (age range, 1 to 80 years; 111 male and 103 female subjects) with middle or inner ear disease to assess the extent of sphenoid aeration. We also determined volumes of the sphenoid sinuses on 1.0- or 1.5-mm reformatted images by integrating the sinus air (< or = -900 HU) area. RESULTS: Sphenoid sinus aeration began as a doublet in the anterior boundary of the sphenoid bone by the age of 5 years, with patients more than 6 years old exhibiting varying degrees of aeration. The aeration on both sides continued to expand until the third decade of life. The maximum average volume was 8.2 +/- 0.5 cm3. Thereafter, the volume decreased gradually, with the average volume in the seventh decade of life being 71% of the maximum level. The aeration of the peripheral portions of the sphenoid bone, such as the pterygoid process, anterior clinoid process, and dorsum sella, occurred predominantly after closure of the spheno-occipital suture, and showed a tendency to recede during aging. CONCLUSION: Volumetric assessment of the sphenoid sinus by helical CT scanning revealed age-related expansion and reduction in aeration. PMID- 10669248 TI - In vivo 1H MR spectroscopy of human head and neck lymph node metastasis and comparison with oxygen tension measurements. AB - BACKGROUND AND PURPOSE: Current diagnostic methods for head and neck metastasis are limited for monitoring recurrence and assessing oxygenation. 1H MR spectroscopy (1H MRS) provides a noninvasive means of determining the chemical composition of tissue and thus has a unique potential as a method for localizing and characterizing cancer. The purposes of this investigation were to measure 1H spectral intensities of total choline (Cho), creatine (Cr), and lactate (Lac) in vivo in human lymph node metastases of head and neck cancer for comparison with normal muscle tissue and to examine relationships between metabolite signal intensities and tissue oxygenation status. METHODS: Volume-localized Lac-edited MRS at 1.5 T was performed in vivo on the lymph node metastases of 14 patients whose conditions were untreated and who had primary occurrences of squamous cell carcinoma. MRS measurements were acquired also from the neck muscle tissue of six healthy volunteers and a subset of the patients. Peak areas of Cho, Cr, and Lac were calculated. Tissue oxygenation (pO2) within the abnormal lymph nodes was measured independently using an Eppendorf polarographic oxygen electrode. RESULTS: Cho:Cr ratios were significantly higher in the nodes than in muscle tissue (node Cho:Cr = 2.9 +/- 1.6, muscle Cho:Cr = 0.55 +/- 0.21, P = .0006). Lac was significantly higher in cancer tissue than in muscle (P = .01) and, in the nodes, showed a moderately negative correlation with median pO2 (r = -.76) over a range of approximately 0 to 30 mm Hg. Nodes with oxygenation values less than 10 mm Hg had approximately twice the Lac signal intensity as did nodes with oxygenation values greater than 10 mm Hg (P = .01). Cho signal intensity was not well correlated with pO2 (r = -.46) but seemed to decrease at higher oxygenation levels (>20 mm Hg). CONCLUSION: 1H MRS may be useful for differentiating metastatic head and neck cancer from normal muscular tissue and may allow for the possibility of assessing oxygenation. Potential clinical applications include the staging and monitoring of treatment. PMID- 10669249 TI - Idiopathic sclerotic inflammation of the orbit with left optic nerve compression in a patient with multifocal fibrosclerosis. AB - We present the MR imaging findings in a 43-year-old male patient with bilateral idiopathic sclerosing inflammation of the orbit. Bilateral enhancing retrobulbar masses, with concentric compression of the retrobulbar segment of the left optic nerve, were seen. MR imaging proved to be the only means to distinguish between the different intraorbital structures and to determine the exact site of optic nerve compression. To our knowledge, this is the first documented case of MR imaging findings of this entity. PMID- 10669250 TI - Optic nerve cysticercosis: imaging findings. AB - We present the imaging findings of retrobulbar optic nerve cysticercosis in a 50 year-old woman with a 6-month history of vision loss. Contrast-enhanced CT revealed an approximately 7-mm ring-enhancing cyst with a mural nodule located in the anterior portion of the left optic nerve. A contrast-enhanced MR imaging study revealed a cystic lesion with peripheral enhancement of the mural nodule. Sonography showed a cystic mass with a focal area of increased echogenicity corresponding to the mural nodule. PMID- 10669251 TI - Thermally induced transient trigeminal sensory neuropathy: imaging findings. AB - We report the clinical and imaging features of a patient with transient partial trigeminal sensory neuropathy thought to have been induced by thermal injury to the tongue. Abnormal thickening and enhancement of the mandibular division of the trigeminal nerve was revealed by MR imaging. The diagnostic considerations for mass-like enlargement of the trigeminal nerve should include transient/inflammatory processes, as well as more common and sinister conditions, such as tumor. PMID- 10669252 TI - Increased detectability of alpha brain glutamate/glutamine in neonatal hypoxic ischemic encephalopathy. AB - BACKGROUND AND PURPOSE: Proton MR spectroscopy (MRS) detectability of brain glutamate/glutamine (Glx) is increased in hypoxic-ischemic insults and is implicated in the neuronal injury and death that follows. Our aim was to correlate the proton MRS detectability of alpha-CH protons of Glx (alpha-Glx) with the Sarnat stage of neonatal hypoxic-ischemic encephalopathy (HIE). METHODS: Initial and follow-up proton MRS studies at 1.9 T were performed in 28 neonates aged 1 to 7 days (seven healthy control subjects and 21 with HIE: 10 mild, nine moderate, and two severe) and in 12 neonates aged 13 to 17 days (12 with HIE: eight mild, three moderate, and one severe), respectively. Both point-resolved spectroscopy (PRESS) and stimulated-echo acquisition mode (STEAM) sequences were used. The spectral volume of interest was in the basal ganglia, thalami, and adjoining regions. The detectability of alpha-Glx was assessed by two different parameters: the detection rate of the alpha-Glx peak and the peak-area ratio of alpha-Glx to creatine and phosphocreatine. RESULTS: On both the initial and follow-up PRESS studies, all the neonates with moderate and severe HIE showed an alpha-Glx peak, compared with one healthy control subject in the initial study and one neonate with mild HIE in both the studies. They also demonstrated a significantly higher peak-area ratio of alpha-Glx/(creatine and phosphocreatine) on both the initial and follow-up studies. The peak-area ratios in neonates with HIE positively correlated with the Sarnat stage of HIE on both the initial and follow-up studies. Neonates with moderate and severe HIE also showed a consistently higher alpha-Glx peak on both the initial and follow-up studies with the STEAM sequence. CONCLUSION: Proton MRS detectability of alpha-Glx is increased in moderate and severe HIE and correlates with the Sarnat stage of HIE. PMID- 10669253 TI - Sonography, CT, and MR imaging: a prospective comparison of neonates with suspected intracranial ischemia and hemorrhage. AB - BACKGROUND AND PURPOSE: Sonography, CT, and MR imaging are commonly used to screen for neonatal intracranial ischemia and hemorrhage, yet few studies have attempted to determine which imaging technique is best suited for this purpose. The goals of this study were to compare sonography with CT and MR imaging prospectively for the detection of intracranial ischemia or hemorrhage and to determine the prognostic value(s) of neuroimaging in neonates suspected of having hypoxic-ischemic injury (HII). METHODS: Forty-seven neonates underwent CT (n = 26) or MR imaging (n = 24) or both (n = 3) within the first month of life for suspected HII. Sonography was performed according to research protocol within an average of 14.4 +/- 9.6 hours of CT or MR imaging. A kappa analysis of interobserver agreement was conducted using three independent observers. Infants underwent neurodevelopmental assessment at ages 2 months (n = 47) and 2 years (n = 26). RESULTS: CT and MR imaging had significantly higher interobserver agreement (P < .001) for cortical HII and germinal matrix hemorrhage (GMH) (Grades I and II) compared with sonography. MR imaging and CT revealed 25 instances of HII compared with 13 identified by sonography. MR imaging and CT also revealed 10 instances of intraparenchymal hemorrhage (>1 cm, including Grade IV GMH) compared with sonography, which depicted five. The negative predictive values of neuroimaging, irrespective of technique used, were 53.3% and 58.8% at the 2-month and 2-year follow-up examinations, respectively. CONCLUSION: CT and MR imaging have significantly better interobserver agreement for cortical HII and GMH/intraventricular hemorrhage and can reveal more instances of intraparenchymal hemorrhage compared with sonography. The absence of neuroimaging findings on sonograms, CT scans, or MR images does not rule out later neurologic dysfunction. PMID- 10669254 TI - Hemangioendothelioma of the parotid gland in infants: sonography and correlative MR imaging. AB - BACKGROUND AND PURPOSE: Hemangioendothelioma is the most common parotid gland tumor of childhood, and is diagnosed on clinical grounds, supported by imaging findings. Previous work has suggested that MR is the best imaging technique for assessment of parotid hemangioendothelioma. Demonstration of a reliable sonographic appearance would reduce the need for MR imaging in infants with this lesion. METHODS: We performed high-frequency sonography (including color Doppler and power Doppler imaging) in three patients, each with a diagnosis of parotid hemangioendothelioma confirmed by clinical follow-up. Two patients were also examined with MR imaging and labeled red cell scintigraphy. RESULTS: All sonographic studies showed a homogeneous mass enlarging and replacing most or all of the visualized parotid gland, with a lobular internal structure, fine echogenic internal septations, and a mildly lobulated contour. Color Doppler and power Doppler imaging showed extremely high vascularity within the mass. Correlative MR images in two infants showed a well-defined lesion with uniform intense contrast enhancement. Labeled red cell scintigraphy showed a well-defined area of intense activity. CONCLUSION: In the presence of a typical clinical history, sonography and clinical follow-up alone may prove to be sufficient for safe management of parotid hemangioendothelioma. MR imaging or labeled red cell scintigraphy may only be required if the sonographic features are atypical. These findings require confirmation in a larger series of patients. PMID- 10669255 TI - Leigh syndrome in a 3-year-old boy with unusual brain MR imaging and pathologic findings. AB - We report unusual MR serial imaging and electron microscopy findings in a 3-year old boy who had Leigh syndrome with cytochrome-c oxidase (cox) deficiency. The MR imaging findings included periventricular white matter involvement, posteroanterior progression, and extension through the corpus callosum and internal capsule; however, no basal ganglia or brain stem abnormality was found, which was suggestive of leukodystrophy. The most noteworthy findings were the cystic foci with contrast enhancement in the affected white matter. PMID- 10669256 TI - In re: Bydder GM, Steiner RE, Young IR, et al. Clinical NMR imaging of the brain: 140 cases. Am J Reontgenol 1982;139:215-236. PMID- 10669257 TI - Internal carotid artery narrowing in children with retropharyngeal lymphadenitis and abscess. PMID- 10669258 TI - In re: Basilar artery migraine and reversible imaging abnormalities. PMID- 10669259 TI - Respiratory viral infections in the elderly. AB - Viral respiratory infections represent a significant challenge for those interested in improving the health of the elderly. Influenza continues to result in a large burden of excess morbidity and mortality. Two effective measures, inactivated influenza vaccine, and the antiviral drugs rimantadine and amantadine, are currently available for control of this disease. Inactivated vaccine should be given yearly to all of those over the age of 65, as well as younger individuals with high-risk medical conditions and individuals delivering care to such persons. Live, intranasally administered attenuated influenza vaccines are also in development, and may be useful in combination with inactivated vaccine in the elderly. The antiviral drugs amantadine and rimantadine are effective in the treatment and prevention of influenza A, although rimantadine is associated with fewer side-effects. Recently, the inhaled neuraminidase inhibitor zanamivir, which is active against both influenza A and B viruses, was licensed for use in uncomplicated influenza. The role of this drug in treatment and prevention of influenza in the elderly remains to be determined. Additional neuraminidase inhibitors are also being developed. In addition, to influenza, respiratory infections with respiratory syncytial virus, parainfluenza virus, rhinovirus, and coronavirus have been identified as potential problems in the elderly. With increasing attention, it is probable that the impact of these infections in this age group will be more extensively documented. Understanding of the immunology and pathogenesis of these infections in elderly adults is in its infancy, and considerable additional work will need to be performed towards development of effective control measures. PMID- 10669260 TI - Effect of Hochu-ekki-to (TJ-41), a Japanese herbal medicine, on the survival of mice infected with influenza virus. AB - The antiviral effect of Hochu-ekki-to (TJ-41), a Japanese herbal medicine, was investigated using mice infected with influenza virus. TJ-41 was found to increase the survival rate, prolong the mean survival days, suppress viral growth in bronchoalveolar labage fluid (BALF) and inhibit the lung index (lung consolidation) on day 4 after infection in mice infected with influenza, after the agent had been administered orally once daily from day 7 to 2 before infection and from day 0 to 4 after infection. Administration of TJ-41 decreased the BALF concentrations of IL-1alpha, IL-6 and GM-CSF, but not TNF-alpha or interferon-gamma (IFN-gamma), on day 4 after infection. In addition, TJ-41 elevated the level of IFN-alpha in BALF on day 2 after infection. Yet, TJ-41 did not show any inhibitory effect on the growth of influenza virus in vitro. These results suggest that TJ-41 exerts its inhibitory effect on influenza virus infection via enhancement of the host immune responses in this experimental murine system. PMID- 10669261 TI - Antiviral properties of a mangrove plant, Rhizophora apiculata Blume, against human immunodeficiency virus. AB - A polysaccharide extracted from the leaf of Rhizophora apiculata (RAP) was assessed in cell culture systems, for its activity against human and simian immunodeficiency viruses. RAP inhibited HIV-1 or HIV-2 or SIV strains in various cell cultures and assay systems. It blocked the expression of HIV-1 antigen in MT 4 cells and abolished the production of HIV-1 p24 antigen in peripheral blood mononuclear cells (PBMC); the 50% effective concentration (EC50) of RAP in HIV-1 infected MT-4 cells and in PBMC was 10.7 and 25.9 microg/ml, respectively. RAP (100 microg/ml) completely blocked the binding of HIV-1 virions to MT-4 cells. RAP also reduced the production of viral mRNA when added before virus adsorption. RAP inhibited syncytium formation in cocultures of MOLT-4 cells and MOLT-4/HIV 1(IIIB) cells. RAP did not prolong activated partial thromboplastin time (APTT) up to 500 microg/ml. These properties may be advantageous should RAP be considered for further development. PMID- 10669262 TI - Inhibitory effect of dibenzofuran and dibenzosuberol derivatives on rhinovirus replication in vitro; effective prevention of viral entry by dibenzosuberenone. AB - A series of derivatives of dibenzofuran and dibenzosuberol block rhinovirus replication in vitro as judged by their ability to hinder the cytopathic effect in cells infected with HRV14 or HRV16. Both the number and the size of viral plaques were reduced effectively by treatment with these compounds in a dose dependent fashion, thus affecting viral spread. The compound 2-hydroxy-3 dibenzofuran carboxylic acid was equally effective against HRV16 and HRV14, with IC50 values of 25 microM in cytopathy assays. Dibenzosuberenone showed minor differences in selectivity, with IC50 values of 10 and 30 microM for HRV16 and HRV14 cytopathy, respectively. Likewise, dibenzosuberenone effectively prevented the production of HRV16 proteins, viral RNA, and infectious virus particles when present at concentrations above 30 microM. Time-of-addition experiments show that compounds must be administered before or during the viral adsorption step in order to be effective antivirals. Dibenzosuberenone can block the adsorption of viral particles on to cells, preventing further steps in the replication cycle, but is not effective as a direct inactivating agent. These compounds likely interact with viral capsid proteins, affecting receptor interactions required for attachment and subsequent entry into cells. PMID- 10669263 TI - Heterotypic inhibition of foot-and-mouth disease virus infection by combinations of RNA transcripts corresponding to the 5' and 3' regions. AB - Strategies to inhibit RNA virus multiplication based on the use of interfering nucleic acids have to consider the high genetic polymorphism exhibited by this group of viruses. Here, we report high levels of heterotypic inhibition of foot and-mouth disease virus (FMDV) infective particle formation in cotransfection experiments of susceptible cell lines with infections viral RNA and combinations of viral transcripts. The interfering molecules used include the following regions on type C FMDV RNA: (i) sequences from the 5' region, spanning the proximal part of the internal ribosome entry site element and the two functional initiator AUGs; and (ii) the 3' terminal region including the 3' end of 3D gene and the complete 3' non-coding region. Combination of 5' antisense RNA molecules with either sense or antisense RNA molecules from the 3' region resulted in inhibition of up to 90% of the infectivity of homologous type C FMDV RNA. The inhibition was dose-dependent and specific, as no reduction was observed in the plaque-forming units recovered from RNA of swine vesicular disease virus, a related picornavirus. Interestingly, high levels-of intertypic inhibition, about 60% or higher, were observed when viral RNAs of serotypes O and A were analysed. These levels of inhibition are consistent with the levels of nucleotide homology exhibited by the viruses analysed in the target sequences. Inhibition of virus yield was also observed in FMDV-infected cells transiently expressing the interfering RNAs. Thus, transcripts of the FMDV RNA corresponding to the 5' and 3' regions specifically inhibit FMDV particle formation in a serotype-independent manner. PMID- 10669264 TI - Anaesthesiology into the new millenium. PMID- 10669265 TI - PaO2 during anaesthesia and years of smoking predict late postoperative hypoxaemia and complications after upper abdominal surgery in patients without preoperative cardiopulmonary dysfunction. AB - BACKGROUND: The incidence of late postoperative hypoxaemia and complications after upper abdominal surgery is 20-50% among cardiopulmonary healthy patients. Atelectasis development during anaesthesia and surgery is the main hypothesis to explain postoperative hypoxaemia. This study tested the predictive value of PaO2<19 kPa during combined general and thoracic epidural anaesthesia and the preoperative functional residual capacity (FRC) reduction in the 30 degree head tilt-down position for the development of late prolonged postoperative hypoxaemia, PaO2<8.5 kPa for a minimum of 3 out of 4 days, and other complications. Forty patients without cardiopulmonary morbidity, assessed by ECG, spirometry, FRC and diffusion capacity preoperatively, underwent upper abdominal surgery. PaO2 during anaesthesia and preoperative FRC reduction were compared to known risk factors for the development of hypoxaemia and complications: age, pack years of smoking and duration of operation. The effect of optimizing pulmonary compliance with peroperative positive end-expiratory pressure (PEEP) on postoperative hypoxaemia and complications was evaluated in a blinded and randomized manner. RESULTS: Late prolonged postoperative hypoxaemia and other complications were found in 37% and 38% of the patients, respectively. Patients with PaO2>19 kPa during anaesthesia with F(I)O2=0.33 exhibited a risk, irrespective of PEEP status, of suffering late prolonged hypoxaemia of 0% (0;23) and patients with PaO2<19 kPa a risk of 52% (32;71), P<0.005. Having smoked more than 20 pack-years was associated with a 47% (19;75) higher incidence of postoperative complications than having smoked less than 20 pack-years, P<0.006. CONCLUSIONS: PaO2 during anaesthesia and smoked pack-years provide new tools evaluating patients undergoing upper abdominal surgery in order to predict the patients who develop late postoperative hypoxaemia and complications. PMID- 10669266 TI - Lipid peroxidation induced by an early inflammatory response in endotoxaemia. AB - BACKGROUND: Endotoxaemic challenge promptly causes lipid peroxidation. Porcine endotoxaemia can be used to replicate severe human septic shock. This model was used to evaluate non-enzymatic [8-Iso-prostaglandin F2alpha (8-Iso-PGF2alpha)] and enzymatic [15-keto-13,14-dihydro-prostaglandin F2alpha (15-K-DH-PGF2alpha)] lipid peroxidation, respectively, in relation to survival. The aim of this study was to correlate, if possible, pathophysiologic events during endotoxaemia to the levels of these arachidonic acid metabolites. METHODS: Nineteen pigs were anaesthetised, monitored (circulatory and respiratory variables in relation to lipid peroxidation) and given a continuous 6 h E. coli endotoxin (10 microg x kg( 1) x h(-1)) infusion. All animals were mechanically ventilated at constant tidal volumes and the inspired oxygen fraction was kept constant during the experimental period. RESULTS: This endotoxin infusion caused expressed derangements in all pigs and death in 9 of them. The levels of 8-Iso-PGF2alpha, indicating oxidative injury, were different in time course, magnitude and fashion between survivors and non-survivors. The levels of 15-K-DH-PGF2alpha, indicating inflammatory response, showed a similar pattern. At 1 h the CO2 partial pressure in arterial blood was significantly higher in non-surviving pigs and correlated (r: 0.7; P<0.05) to the levels of 15-K-DH-PGF2alpha. Prostaglandin F2alpha is mainly metabolised in the lung. The lung weights were significantly (P<0.05) higher in non-surviving than in surviving animals. Both free radical and cyclooxygenase catalysed oxidative modification occurs during endotoxaemia. CONCLUSION: Increased metabolism and inflammation, as evaluated by 15-K-DH PGF2alpha, in the group of non-survivors may mediate the increase in arterial CO2. Thus, increased lipid peroxidation seems to be associated with endotoxaemic organ dysfunction and increased mortality. PMID- 10669267 TI - Effects of melagatran, an inhibitor of thrombin, on fibrin deposits, haemodynamics, and platelet count in endotoxaemic pigs. AB - BACKGROUND: Thrombin plays a pivotal role in the development of septic shock. Porcine endotoxaemia can replicate this condition. We wanted to evaluate whether melagatran, a novel inhibitor of thrombin, would counteract some of the endotoxin induced changes in this model. METHODS: Fifteen pigs were anaesthetised, monitored (circulatory and respiratory variables) and subjected to an infusion of E. coli endotoxin at 10 microg x kg(-1) x h(-1). Six pigs were given melagatran during the first 3 h of the 6-h endotoxaemic period. Nine controls were given the corresponding volume of saline instead of melagatran. Specimens from the liver and the left lung were taken for light microscopy post mortem. RESULTS: The endotoxin-induced increase in pulmonary capillary wedge pressure and drop in platelet count were significantly less pronounced in the melagatran-treated pigs. Deposits of pulmonary fibrin were significantly reduced in the melagatran group, without improving oxygenation. Light microscopy revealed no hepatic fibrin in the pigs treated with melagatran in contrast to the endotoxaemic controls. Hepatic neutrophil accumulation was reduced in the melagatran group as compared to controls. Hepatocellular degeneration and plasma levels of tumour necrosis factor alpha (TNF alpha) and bilirubin were of the same magnitude in both groups. CONCLUSION: Melagatran reduced pulmonary capillary wedge pressure, a retrograde reflection of the left ventricular end-diastolic filling pressure, and also pulmonary stasis in pigs subjected to endotoxaemic challenge. Pulmonary and hepatic fibrin depositions were reduced, but PaO2 levels or liver function markers were not affected by melagatran during the early phase of endotoxaemia. Obstruction of the intrahepatic bile ducts, by fibrin depositions, is not responsible for reduced excretion of conjugated bilirubin during endotoxaemia. The beneficial effects of melagatran during endotoxaemia were not due to any reduction of plasma TNF alpha. PMID- 10669268 TI - Dual mode antimony electrode for simultaneous measurements of PO2 and pH. AB - BACKGROUND: In biomedical research and clinical medicine there is a demand for potent sensors to measure the components that make up blood gas analyses. Today, as when the electrochemical PO2, PCO2 and pH electrodes were first introduced, these measurements are usually made with the same type of sensor technology. The aims of the present study were, firstly, to find out whether the platinum cathode in the Clark electrode can be replaced by antimony for oxygen measurements (amperometry (A)); secondly, whether, during oxygen measurements, the inherent corrosion potential of the antimony metal can be used for measurement of pH in the same measurement area (potentiometry (P)). METHODS: An electrode of purified, crystallographically orientated monocrystalline antimony (COMA) connected to a reference electrode (silver-silver chloride) was used for the P measurements. Measurements of A (at -900 mV) and P were made in an aqueous environment regulated for oxygen, pH, and temperature. RESULTS: Reproducible oxygen sensitivities of 0.925 nA/% oxygen (2% CV (coefficient of variation)) (A), 10.7 mV/% (P), and 0.7 mV/% (P) were found in the oxygen range: 0-21%, <5%, and above 5%, respectively. The pH sensitivity was 57 mV/pH unit (P). Oxygen and pH measurements were less accurate at oxygen concentrations close to 0%. CONCLUSIONS: Both the oxygen and pH part of the composite electrode signal can be identified by this dual mode technique (A and P). The sensor seems to be promising as it provides measurements of two separate variables (oxygen and pH) and also has the desirable characteristics of a solid state sensor. PMID- 10669269 TI - Continuous epidural analgesia with bupivacaine-fentanyl versus patient-controlled analgesia with i.v. morphine for postoperative pain relief after knee ligament surgery. AB - BACKGROUND: Both epidural analgesia and intravenous patient-controlled analgesia (PCA) have been found efficacious after various types of surgery. We compared the efficacy, safety, side effects and patient satisfaction of these methods in a randomized double-blind fashion after elective anterior cruciate ligament reconstruction of the knee. METHODS: Fifty-six patients had an epidural catheter placed at the L2-L3 interspace. Spinal anaesthesia with 15 mg of plain bupivacaine 5 mg/ml was performed at the L3-L4 interspace. After surgery the patients were randomly divided into three groups: 19 received a continuous epidural infusion with bupivacaine 1 mg/ml and fentanyl 10 mg/ml (F10), 19 patients received bupivacaine 1 mg/ml and fentanyl 5 microg/ml (F5) and 18 patients received saline (S). The rate of the epidural infusions was 0.1 ml kg( 1) h(-1). Each patient could also use an intravenous (i.v.) PCA device with 40 microg/kg bolus doses of morphine with a lockout period of 10 min and a maximum dose 240 microg kg(-1) h(-1). At the end of surgery ketoprofen 100 mg i.v. was given and continued orally three times a day. Patients were assessed for pain with a visual analogue scale (VAS) at rest and during activity, side effects and satisfaction at 3, 9 and 20 h. RESULTS: Both epidural infusions (F10, F5) provided better analgesia than epidural saline plus i.v. PCA (S) (P<0.05). There was slightly less nausea in the S group (NS). In spite of the difference in the quality of pain relief, there was no difference between the groups in patient satisfaction regarding analgesic therapy. CONCLUSION: Epidural infusion of fentanyl (1 microg kg(-1) h(-1) or 0.5 microg kg(-1) h(-1)) and bupivacaine (0.1 mg kg(-1) h(-1)) provided better pain relief but more side effects than intravenous morphine patient-controlled analgesia after knee ligament surgery. Almost all patients in all groups were satisfied with their pain relief. PMID- 10669270 TI - Patient-controlled versus nurse-controlled pain treatment after coronary artery bypass surgery. AB - BACKGROUND: Pain after coronary artery bypass surgery persists for several days. A continuous intravenous infusion of an opioid adequately accomplishes good pain control in the intensive care unit, but it is often not suitable on the ordinary ward. Patient-controlled analgesia (PCA) with intermittent injections delivered by one of the new devices now available could be an alternative to conventional nurse-controlled analgesia (NCA) based on intermittent injections. The aim was to compare these two techniques with respect to efficacy and the amount of opioid used. METHODS: Forty-eight patients randomly received PCA or NCA with ketobemidone following extubation after coronary artery bypass grafting. Drug consumption, pain assessment with the visual analogue score (VAS) and possible side effects were evaluated from extubation to the end of the second postoperative day. RESULTS: On the day of surgery the VAS scores did not differ between the groups. From the afternoon of the first postoperative day the VAS scores were higher in the NCA group with mean values at 3-4 out of 10 as compared with mean values around 2 in the PCA group (P<0.01). During the study period the patients in the PCA group received more ketobemidone as compared with the NCA group, 61.9+/-24.0 mg and 36.3+/-20.2 mg, respectively (P<0.01). Additional oral analgesics were used in 12 of the patients in the NCA group compared with none in the PCA group. The few side effects reported were equally distributed between the two groups. CONCLUSION: PCA treatment after coronary artery bypass surgery resulted in better pain treatment and the use of more opioid without an increase in side effects compared with traditional NCA treatment. PMID- 10669271 TI - Bradycardia and asystolic cardiac arrest during spinal anaesthesia: a report of five cases. AB - Sudden, severe bradycardia/asystolic cardiac arrest are considered infrequent, but are certainly the most serious complications of spinal anaesthesia. We report four cases of primary asystole and one of severe bradycardia in young to middle aged, healthy patients scheduled for minor surgery at the day surgery unit. Bradycardia/asystole were not related to respiratory depression or hypoxaemia/hypercarbia; they occurred at different time intervals after the onset of spinal anaesthesia (10-70 min) and, apparently, were not dependent on the level of sensory block, which varied between T3 and T8. One patient was nauseated seconds before the asystole, otherwise there was no warning signs. All the patients were easily resuscitated with the prompt administration of atropine and ephedrine and, in the case of cardiac arrest, cardiac massage and ventilation with oxygen. One patient was treated with a small dose of adrenaline. Four patients had the surgery, as planned; one had the surgery postponed. All the patients were discharged from hospital in good health and did not suffer any sequelae. PMID- 10669272 TI - The effects of clonidine on ropivacaine 0.75% in axillary perivascular brachial plexus block. AB - INTRODUCTION: The new long-acting local anesthetic ropivacaine is a chemical congener of bupivacaine and mepivacaine. The admixture of clonidine to local anesthetics in peripheral nerve block has been reported to result in a prolonged block. The aim of the present study was to evaluate the effects of clonidine added to ropivacaine on onset, duration and quality of brachial plexus block. METHODS: Patients were randomly allocated into two groups. In group I brachial plexus was performed using 40 ml of ropivacaine 0.75% plus 1 ml of NaCL 0.9%, and in group II brachial plexus was performed using 40 ml of ropivacaine 0.75% plus 1 ml (0.150 mg) of clonidine. Onset of sensory and motor block of radial, ulnar, median and musculocutaneous nerve were recorded. Motor block was evaluated by quantification of muscle force, according to a rating scale from 6 (normal contraction force) to 0 (complete paralysis). Sensory block was evaluated by testing response to a pinprick in the associated innervation areas. Finally, the duration of the sensory block was registered. Data were expressed in mean+/-SD. For statistical analysis a Student t-test was used. A P-value of < or = 0.05 was considered as statistically significant. RESULTS: The duration of blockade was without significant difference between the groups. Group I: 718+/-90 min; Group II: 727+/-117 min. There was no intergroup difference in sensory and motor onset or in quality of blockade. CONCLUSION: The addition of clonidine to ropivacaine 0.75% does not lead to any advantage of block of the brachial plexus when compared with pure ropivacaine 0.75%. PMID- 10669273 TI - Pain intensity and pain relief after surgery. A comparison between patients' reported assessments and nurses' and physicians' observations. AB - BACKGROUND: Postoperative pain remains a problem for many patients. One of the reasons could lie in the insufficient evaluation of pain and analgesia. This study was designed to obtain more insight in the performance of nurses and physicians in evaluating patients' postoperative pain and pain relief. METHODS: Forty patients hospitalised in one surgical unit and the 8 nurses and the 2 surgical residents in charge of this unit were investigated. Patients were asked to assess on a visual analogue scale the intensity of their pain and their pain relief at rest, on coughing and globally since the operation, on the first and second postoperative days and the day before hospital discharge. Separately, the nurses and the physicians were asked to evaluate the pain intensity and the pain relief for each patient involved. A MANOVA and a multiple comparisons test with Bonferroni adjustment were used. RESULTS: At rest, only nurses underestimated pain intensity on the day before hospital discharge. On coughing, physicians underestimated pain intensity in all 3 assessments, whereas nurses only in the 3rd assessment (on the day before hospital discharge). Globally, physicians underestimated pain intensity in all 3 assessments, nurses in the 2nd and the 3rd assessment. Only physicians overestimated pain relief on coughing on the day before hospital discharge and globally in all 3 assessments. Surprisingly, the pain scores rated by the patients before hospital discharge were high. CONCLUSION: The results of this survey suggest that assessment of pain and pain relief is inadequately done by both physicians and nurses. This emphasises the importance of a better training, and a systematic assessment of pain intensity and pain relief. PMID- 10669274 TI - Pre-incisional epidural ketamine, morphine and bupivacaine combined with epidural and general anaesthesia provides pre-emptive analgesia for upper abdominal surgery. AB - BACKGROUND: Previous studies have shown that N-methyl-D-asparate (NMDA) receptor antagonists provide a pre-emptive analgesic effect in humans. This study investigated the benefits of pre-emptive analgesia for upper abdominal surgery, using pre-incisional epidural ketamine + morphine + bupivacaine (K+M+B) treatment for achieving postoperative pain relief. METHODS: Sixty ASA 1-2 patients scheduled for upper abdominal surgery were allocated to three groups in a randomized, single-blinded study. Patients in the control group (I) received general anaesthesia followed by an infusion of normal saline. Group II and III patients received general anaesthesia with a continuous epidural infusion of 2% lidocaine. Thirty minutes after the incision in groups I and II, an epidural pain control regimen was administered using ketamine (10 mg) and morphine (1 mg) in 10 ml of 0.085% bupivacaine (K+M+B). Group III patients also received K+M+B, but it was administered 10 min after the 2% lidocaine injection and 30 min before skin incision. All patients received an epidural pain control regimen (q12 h) for 3 days after their first injection. Patient-controlled analgesia (PCA) with morphine was used to control subsequent postoperative pain. During the 3-day period following surgery, duration to PCA trigger (h), morphine consumption (mg), pain intensity at rest and when coughing/moving, and analgesic-related adverse effects were recorded. The VAS scale (0-10) was used to assess pain intensity. RESULTS: Median times to first PCA trigger were 1.2 (0.5-2.0) h, 3.0 (0.7-4.2) h, and 4.0 (2.5-7.5) h for groups I, II, and III, respectively. Both the incident and resting pain scores were consistently lower for group III patients than groups I and II. The number of PCA triggers (all attempts/successful triggers) during the day following surgery were 14.0 (3-30)/8.0 (3-24) times, 10.0 (3 23)/6.0 (2-20) times, and 7.0 (3-12)/4.5 (1-10) times for groups I, II, and III. Total morphine consumption for the 3-day observation period was 12.5 (3-42) mg, 10.5 (2-29) mg, and 6.0 (1-20) for groups I, II, and III, respectively. CONCLUSION: Pre-incisional epidural K+M+B treatment combined with continuous epidural anaesthesia and general anaesthesia provides an ideal pre-emptive analgesic therapy, exhibiting better postoperative pain relief than general anaesthesia and post-incisional K+M+B treatment. PMID- 10669275 TI - Platelet dysfunction after intravenous ketorolac or propacetamol. AB - BACKGROUND: Paracetamol is a weak cyclo-oxygenase inhibitor in vitro. A recent study in children has shown that high doses of paracetamol are effective and safe. We studied the effect of propacetamol on haemostasis in adult volunteers. METHODS: Ten volunteers were investigated in a double-blind, randomized, crossover study. They received propacetamol 60 mg kg(-1) or ketorolac 0.4 mg kg( 1) in saline i.v. (30 min) in two different sessions. Platelet function was evaluated before the test infusion (S-0), two (S-2) and 24 h (S-24) after the start of the infusion. Coagulation parameters (PT, APTT, factor V and VII activities) were measured at S-0, S-24 and 48 h (S-48). RESULTS: One of the volunteers had no secondary platelet aggregation in S-0 and was excluded from the final analysis. Two hours (S-2) after propacetamol and ketorolac administration the adrenaline (0.9 microg ml(-1) and 9.0 microg ml(-1)) induced maximal platelet aggregation was decreased compared with S-0. At S-2 platelet aggregation was inhibited more after ketorolac than after propacetamol. At 24 h after ketorolac, but not after propacetamol, there was still a decrease in the adrenaline-induced maximal platelet aggregation. Propacetamol did not affect adenosine diphosphate (ADP)-induced maximal platelet aggregation, whereas ketorolac decreased 3 and 6 microM ADP-induced maximal platelet aggregation at S-2 and S-24. However, 2 h after both ketorolac and propacetamol, thromboxane B2 (TxB2) concentration decreased in platelet rich plasma after 5 min aggregation induced by 8 microM ADP. Coagulation was unaffected. CONCLUSION: Propacetamol 60 mg kg(-1) i.v. causes reversible platelet dysfunction demonstrated by a decrease in maximal platelet aggregation and TxB2 concentration. After 0.4 mg kg(-1) ketorolac i.v. platelet aggregation and TxB2 formation are inhibited more in comparison with propacetamol, and platelet dysfunction is still seen after 24 h. PMID- 10669276 TI - Effects of conventional physiotherapy, continuous positive airway pressure and non-invasive ventilatory support with bilevel positive airway pressure after coronary artery bypass grafting. AB - BACKGROUND: Coronary artery bypass graft (CABG) surgery with the use of mammary arteries is associated with severe alteration of lung function parameters. The purpose of the present study was to compare the effect on lung function tests of conventional physiotherapy using incentive spirometry (IS) with non-invasive ventilation on continuous positive airway pressure (CPAP) and with non-invasive ventilation on bilevel positive airway pressure (BiPAP or NIV-2P), METHODS: Ninety-six patients were randomly assigned to 1 of 3 groups: NIV-2P (1 h/3 h), CPAP (1 h/3 h) and IS (20/2 h). Pulmonary function tests and arterial blood gases analyses were obtained before surgery. On the 1st and 2nd postoperative days, these parameters were collected together with cardiac output and calculation of venous admixture. RESULTS: For the 3 groups a severe restrictive pulmonary defect was observed during the 1st postoperative day. On the 2nd postoperative day, in opposition to IS, intensive use of CPAP and NIV-2P reduced significantly the venous admixture (P<0.001) and improved VC, FEV1 and PaO2 (P<0.01). CONCLUSION: We conclude that preventive use of NIV can be considered as an effective means to decrease the negative effect of coronary surgery on pulmonary function. PMID- 10669277 TI - Dysregulation of immune response following neurosurgical operations. AB - BACKGROUND: Postoperative infections are common and potentially fatal complications in neurosurgical intensive care medicine. An impairment of immune function has been described after central nervous system surgery and in patients harboring malignant brain tumors. The aim of our study was to investigate whether differences in cell-mediated immunity can be found in patients undergoing craniotomy for surgery of glioblastoma or clipping of an intracerebral aneurysm. METHODS: In order to determine the influence of the underlying disease on the immune system, we measured changes in cytokine concentrations (IL-6, IL-10, TGF beta1) and lymphocyte-subsets (CD3+, CD3+HLA-DR+, CD4+, CD8+, CD19+, and CD16+56+) in 8 patients with glioblastoma and in 8 patients with an intracerebral aneurysm before, during and after the neurosurgical procedure. RESULTS: In the comparison of glioblastoma and aneurysm patients, we could show that IL-6 plasma levels were pre- and intraoperatively higher in the aneurysm-group (P<0.05), and the plasma concentrations of IL-10 and TGF-beta were significantly elevated in the glioma-group. The lymphocyte-subsets showed a significantly lower percentage of NK-cells and activated T-cells in the glioma-group. CONCLUSION: Our results document a significant dysregulation of immune response in glioma patients. This may be induced by elevated plasma concentrations of immunoinhibiting cytokines IL 10 and transforming growth factor-beta 1. PMID- 10669278 TI - Effect of pentaglobin and piperacillin on survival in a rat model of faecal peritonitis: importance of intervention timings. AB - BACKGROUND: Faecal peritonitis is a progressive pathophysiological condition which may lead to multiple organ failure and death. The reason for the associated morbidity and mortality could be attributed to the fact that some of the subtle alterations in cellular function that occur during the early stage of peritonitis are unidentified and consequently missed, leading to inadequate or delayed intervention. Recent studies have shown that early treatment with antibiotic and antisera containing antibodies to lipopolysaccharide (immunoglobulin) improve the survival rate in these patients. The present investigation was undertaken to study the effect of pentaglobin and piperacillin with particular attention to time lag of drug intervention on animal survival following experimental peritonitis. METHODS: Experimental peritonitis was produced by inoculating 1 ml/kg of faecal suspension (2:1 w/v in saline) into the peritoneal cavity. Two groups of animals were treated with pentaglobin (4 ml/kg) or piperacillin (1000 mg/kg) respectively, whereas rats in another group received both drugs simultaneously. The first dose of each drug was given at 4 h, 6 h, 8 h and 12 h after faecal inoculation followed by 3 additional doses at 8-h intervals. For biochemical studies, separate groups of animals were used in which the treatment was started 4 h after faecal inoculation and the animals were killed at 12 h after the treatment. RESULTS: Both piperacillin and pentaglobin prolonged survival time of animals which received the treatment within 6 h of faecal insult. The combination of pentaglobin plus piperacillin produced better results as compared to the individual effect. There was a significant decrease in serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and increase in catalase following faeces-induced septicaemia, suggesting a significant increase in oxidative stress. The changes in enzyme levels were significantly attenuated by both the drugs. CONCLUSION: The findings suggest that intervention with a combination of pentaglobin and antibiotics within 6 h of peritonitis might significantly improve survival rate in rat. PMID- 10669279 TI - Nitric oxide does not play a major role in the regulation of systemic hemodynamic responses to acute normovolemic hemodilution. AB - BACKGROUND: The mechanisms of cardiovascular changes following acute normovolemic hemodilution (ANH) have not been fully elucidated. We tested the hypothesis that inhibition of nitric oxide synthesis attenuates ANH-induced cardiovascular responses. METHODS: We observed the effects of N(omega)-nitro-L-arginine methyl ester (L-NAME) pretreatment on ANH-induced cardiovascular responses and compared these effects with those elicited by phenylephrine (PHE). Twenty dogs anesthetized with isoflurane were divided into two groups: one group was pretreated with L-NAME and the other with PHE. Both groups were normovolemically hemodiluted using 6% hydroxyethyl starch to reduce the hemoglobin concentration to approximately 50% of the pretreatment value. RESULTS: Pretreatment with either L-NAME or PHE caused a significant increase in mean aortic blood pressure (MAP) and systemic vascular resistance (SVR) with a significant decrease in cardiac output (CO) and stroke volume (SV). However, no remarkable differences in these variables were seen between groups. In both groups ANH produced increases in heart rate, CO, SV, and maximal left ventricular dP/dt with a significant decrease in SVR. No significant differences in these variables were apparent after ANH except that MAP was decreased in the PHE group but not in the L-NAME group. CONCLUSION: Our results suggest that nitric oxide does not play a major role in mediation or modulation of the systemic vascular responses to ANH. PMID- 10669280 TI - Recovery characteristics of sevoflurane or halothane for day-case anaesthesia in children aged 1-3 years. AB - BACKGROUND: Our objective was to compare the recovery characteristics of sevoflurane and halothane for short day-case anaesthesia in a specifically limited age group of children 1-3 yr. METHODS: Eighty unpremedicated children undergoing day-case adenoidectomy were randomly assigned to receive inhalational induction with either sevoflurane 8% or halothane 5% and nitrous oxide in oxygen (70/30) via a face mask. Tracheal intubation was performed without a muscle relaxant. Anaesthesia was continued with the volatile anaesthetic, adjusted to maintain heart rate and blood pressure within +/-20% of initial values. Recovery was evaluated using a modified Aldrete score, a Pain/Discomfort scale and by measuring recovery end-points. A postoperative questionnaire was used to determine the well-being of the child at home until 24 h after discharge. RESULTS: Emergence and interaction occurred significantly earlier after sevoflurane than halothane but discharge times were similar. More children in the sevoflurane group achieved full Aldrete scores within the first 30 min after anaesthesia, although this group suffered more discomfort during the first 10 min. The amount of postoperative analgesic administered was higher and the first dose given earlier in the sevoflurane group. Postoperative vomiting was more common with halothane, but side-effects in the two groups were otherwise similar in the recovery room and at home. CONCLUSIONS: In children 1-3 yr, sevoflurane provided more rapid early recovery but not discharge after anaesthesia of <30-min duration. Apart from more vomiting with halothane and more discomfort during the first 10 min after awakening with sevoflurane, the quality of recovery was similar with the two anaesthestics. PMID- 10669281 TI - Intramuscular ephedrine reduces emesis during the first three hours after abdominal hysterectomy. AB - BACKGROUND: We tested the hypothesis that intramuscularly administered ephedrine prevents postoperative nausea and vomiting. Ephedrine is cheap, and for this indication poorly documented. METHODS: One hundred and nine patients undergoing elective abdominal hysterectomy under general anaesthesia were studied in a randomized, double-blind placebo-controlled study. Ten minutes before the end of the procedure patients received either ephedrine 0.5 mg/kg i.m. or placebo. The patients were closely observed for 24 h for postoperative nausea or vomiting (PONV) and received a standardized two-step antiemetic treatment of i.v. metoclopramide 10 mg, supplemented with ondansetron 4 mg i.v. if needed. RESULTS: The ephedrine treated patients had significantly less nausea, retching and vomiting, and need of antiemetic rescue during the first 3 h postoperatively compared with the placebo patients. No difference between the groups was evident in the 3-24 h postoperative observation period. All the patients with PONV during 0-3 h experienced PONV in the 3-24 h period. Treatment or prophylaxis with one drug was less efficient than two or more drugs combined. No significant differences in hypotension, tachycardia or other side-effects between the groups were noted. CONCLUSION: Ephedrine 0.5 mg/kg i.m. administered at the end of abdominal hysterectomy has a significant antiemetic effect during the first 3 h after administration with no evident side-effects. PMID- 10669282 TI - Age-related modifications of effects of ketamine and propofol on rat hippocampal acetylcholine release studied by in vivo brain microdialysis. AB - BACKGROUND: We sometimes encounter impairment of learning and memory after general anesthesia in elderly patients. The aim of this study was to examine age related modifications of the effects of ketamine and propofol on rat hippocampal acetylcholine (ACh) release because hippocampal cholinergic neurons are supposed to be involved in learning and memory. METHODS: The experiments were performed on male Wistar young rats (2 months old) and old rats (18 months old), using in vivo brain microdialysis technique under freely moving condition. After initial sampling of three collections, test drugs were administered. The ACh release was determined by the HPLC-ECD method. RESULTS: In old rats, the hippocampal basal ACh release was significantly lower than in young rats. Ketamine (25 and 50 mg kg(-1) i.p.) increased and propofol (25 and 50 mg kg(-1) i.p.) decreased the hippocampal ACh release in both young and old rats. Furthermore, ketamine 50 mg kg(-1) i.p. (anesthetic dose) produced facilitatory effects on the hippocampal ACh release in young rats (193% of the basal release), while in old rats the same dose of ketamine i.p. produced more pronounced facilitatory effects on the hippocampal ACh release (317% of the basal release). On the other hand, propofol 50 mg kg(-1) i.p. (anesthetic dose) produced inhibitory effects on the hippocampal ACh release in young rats (56% of control) and in old rats (77% of control). Although the maximal inhibitory peak effects of propofol 50 mg kg(-1) i.p. did not differ significantly between young rats and old rats, decrease of the hippocampal ACh release in old rats persisted longer than in young rats. CONCLUSION: Ketamine produced more pronounced facilitatory effects on the hippocampal ACh release in old rats, as compared with young rats. On the other hand, propofol has inhibitory effects on the hippocampal ACh release in young and old rats. The aging process may suppress the ability to recover from the inhibitory anesthetic state induced by propofol. PMID- 10669284 TI - Intermittent atrial level right-to-left shunt with temporary hypoxemia in a patient during support with a left ventricular assist device. AB - We report a 56-year-old male patient developing hypoxemia after surgical replacement of infected valves of a left ventricular assist device (LVAD, Novacor) which had supported him during the previous 15 months. Contrast transesophageal echocardiography (TEE) revealed an atrial septal defect with intermittent right-to-left shunt across a patent foramen ovale. We postulate that the shunt detected in this patient occurred as a consequence of reduced pulmonary vascular compliance due to positive end-expiratory pressure (PEEP) and an increase of mean intrathoracic pressure. Furthermore, we hypothesize that synchronized LVAD operation exacerbates any potential right-to-left shunt due to the profound left ventricular unloading which occurs during LVAD support. In this first report of a right-to-left shunt from a previously unrecognized patent foramen ovale in a Novacor patient, the subsequent transient hypoxemia could be managed by avoiding PEEP of more than 3 mmHg, and mean airway pressure of more than 11 mmHg and by careful volume replacement in order to prevent the pump from completely emptying the left ventricle (LV) and the left atrium (LA). Thus, prior to every LVAD implantation a transesophageal contrast echocardiography with Valsalva maneuver should be performed to identify intracardiac right-to-left shunt. PMID- 10669283 TI - Efficacy and safety of premedication with oral ketamine for day-case adenoidectomy compared with rectal diazepam/diclofenac and EMLA. AB - BACKGROUND: Because of its pain-attenuating and sedative properties oral ketamine has been used as premedication in children and adults. We wanted to compare in children scheduled for adenoidectomy safety and efficacy of oral ketamine with a premedication that causes similar preoperative sedation and relief of pain at the venepuncture site. We also evaluated the effect of i.v. glycopyrrolate added to these combinations. METHODS: One hundred children between 10 and 15 kg of body weight scheduled for day-case adenoidectomy were randomly assigned to one of four groups: groups DG and DS received diclofenac 12.5 mg and diazepam 0.5 mg/kg rectally, EMLA cream at the venepuncture site, and placebo orally; groups KG and KS received ketamine 6.0 mg/kg orally, placebo cream at the puncture site, and placebo rectally; additionally, groups DG and KG received glycopyrrolate 5 microg/kg, and groups DS and KS received placebo intravenously. We recorded perioperatively scores (open scale 1-9) for stridor, sedation, bleeding, nausea, pain, heart rate, the need for analgesics and registered psychotomimesis and well being at home. RESULTS: The children of the K-groups became more tearful during separation from their parents (P=0.0072). No other differences were found between the ketamine and diazepam/diclofenac groups before and after premedication until induction of anaesthesia. Oral ketamine produced unpleasant psychotomimesis in four out of 59 children. During the first 10 min postoperatively, the score for stridor was significantly higher in group KS than in the D-groups; stridor scores > or = 6 were seen in one child of the D-groups (DS) and in six children of the K groups (n.s.), of whom three developed laryngospasm (one reintubation). Glycopyrrolate diminished salivation in all groups, but had no effect on stridor scores. Additionally, glycopyrrolate delayed the onset of eating at home. CONCLUSION: Premedication with racemic oral ketamine 6 mg/kg does not seem to be suitable for upper airway procedures. Addition of i.v. glycopyrrolate before the induction of anaesthesia significantly reduced the scores for salivation. PMID- 10669285 TI - Use of solid-phase microextraction for measuring oil-water partition coefficients and correlation with high-performance liquid chromatographic methods for lipophilicity. AB - For flavour compounds, lipophilicity is often estimated by the partition coefficient between oil and water (log Koil-water), which is highly relevant to food. A modification of the shake-flask method is reported here where compounds are quantified in the two phases using solid-phase microextraction (SPME). SPME's highly sensitivity to non-polar compounds facilitates quantification in the water phase. Twelve flavour compounds representing a broad range of lipophilicities and functional groups were analysed by two methods. Their log Koil-water was determined using SPME quantitation and their log k(w) using a reversed-phase HPLC methodology. The isocratic capacity factor at 60% methanol and predicted log P value also showed high correlation factors with other methods. The octadecyl silylated surface of the HPLC column provides a matrix that interacts with lipophilic compounds where the retention time is the indication of lipophilicity. Both methods gave reproducible results (median 3% and 4% RSD) and similar but not identical values for lipophilicity. The relationship between the two methods is log k(w) =0.85 log Koil-water +0.48 with a correlation coefficient of 0.94. The new SPME detection method, with the ability to quantify limonene and 2 pentylfuran at 1 ppm in the water phase, is preferred for flavour compound analysis due to the applicability of oil-water partitioning in food. PMID- 10669286 TI - Liquid chromatography-mass spectrometry method for the determination of aldehydes derivatized by the Hantzsch reaction. AB - A liquid chromatography-mass spectrometry method for the determination of aliphatic aldehydes after derivatization with acetylacetone or dimedone by means of the Hantzsch reaction is presented. Two molecules of a beta-diketone, one molecule of ammonia and an aliphatic aldehyde cyclisate under formation of colored and fluorescent reaction products. Atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) in the positive mode are suitable to ionize the formed dihydropyridine and decahydroacridine derivatives under protonation of their basic secondary amine functionality. The method has been used to identify the oxidation product of the formaldehyde derivatives as side product. The acetaldehyde derivative, presumably formed in the reaction of residual acetaldehyde in the acetic acid or acetate reagents, is mainly responsible for the increasing fluorescence background of the reagent solutions. PMID- 10669287 TI - Molecular mass distribution of sodium alginate by high-performance size-exclusion chromatography. AB - A sensitive high-performance size-exclusion chromatography (HPSEC) method with simple UV detection was developed for the molecular mass analysis of sodium alginate. It was used to evaluate alginates of varying molecular mass and the results were compared with the viscosity measurements. This HPSEC method was sensitive to serve as the stability indicating method for alginate after storage under different conditions. The information of relative molecular mass distribution of alginate was provided with reference to pullulan molecular mass standards. The comparison of the HPSEC chromatograms of alginate, pullulan and dextran revealed the effect of chemical composition of a polysaccharide and its effect on apparent molecular mass distribution. PMID- 10669288 TI - High-performance liquid chromatography method for the characterization of rhamnolipid mixtures produced by pseudomonas aeruginosa UG2 on corn oil. AB - A HPLC method was developed to quantify rhamnolipid species in a bacterial biosurfactant mixture. The biosurfactant mixtures containing mainly 3-[3'-(L rhamnopyranosyl-oxy)decanoyloxy]decanoic acid (RhC10C10), 3-[3'-(2'-O-alpha-L rhamnopyranosyl-oxy)decanoyloxy]decanoic acid (Rh2C10C10), 3-[3'-(2'-O-alpha-L rhamnopyranosyl-oxy)decanoyloxy]dodecanoic acid (Rh2C10C12), and a dehydrogenated variety of the latter, 3-[3'-(2'-O-alpha-L-rhamnopyranosyl oxy)decanoyloxy]dodecenoic acid (Rh2C10C12-H2), were isolated from Pseudomonas aeruginosa UG2 cultures grown on corn oil as sole carbon. The rhamnolipid species were identified and quantified after their derivatization to the corresponding phenacyl esters. To confirm the reliability of the HPLC method, the biosurfactant mixtures and the HPLC isolated species were further analyzed. Mass spectroscopy (electrospray ionization and atmospheric pressure chemical ionization techniques) was used to confirm their molecular mass, gas chromatography to verify their fatty acid content, and a colorimetric assay to quantify the rhamnose content. PMID- 10669289 TI - Determination of octanol-water partition coefficient for terpenoids using reversed-phase high-performance liquid chromatography. AB - Octanol-water partition coefficients (Kow) for 57 terpenoids were measured using a RP-HPLC method. Sample detection was achieved with standard UV and refractive index detectors and required no special column treatment. Measured log Kow values for the terpenoids ranged from 1.81 to 4.48 with a standard error of between 0.03 and 0.08 over the entire range. Partition coefficients determined by the RP-HPLC method were compared against shake flask, atom/fragment contribution, fragment and atomistic methods. The HPLC values were found to give the best correlation with shake flask results. Log Kow values calculated by the atom/fragment contribution method gave the best correlation with the HPLC values when compared to fragment and atomistic methods. PMID- 10669290 TI - Considerations of sample application and elution during size-exclusion chromatography-based protein refolding. AB - A mechanism for size-exclusion chromatography-based protein refolding is described. The model considers the steps of loading the denatured protein onto a gel filtration column, and protein elution. The model predictions are compared with results of refolding lysozyme (10 and 20 mg/ml) using Superdex 75 HR. The main collapse in protein structure occurred immediately after loading, where the partition coefficient of unfolded lysozyme increased from 0.1 to 0.48 for the partially folded molecule. Use of a refolding buffer as the mobile phase resulted in complete refolding of lysozyme; this eluted at an elution volume of 15.6 ml with a final partition coefficient of 0.54. The model predicted the elution volume of refolded lysozyme at 19.3 ml. PMID- 10669291 TI - Eliminating disulfide exchange during glutamyl endopeptidase digestion of native protein. AB - Numerous advantages of using immobilized enzymes over free-solution protein digests have been cited in the literature. This investigation examines both the rate of hydrolysis and the extent of disulfide bond exchange in disulfide bridged dipeptide fragments formed during proteolysis of native protein. Glutamyl endopeptidase as both an immobilized enzyme and in free solution was used in these studies. It was found that extensive hydrolysis of insulin was achieved in 2 min with immobilized enzyme cartridges operated in the stopped-flow mode orders. This is orders of magnitude faster than was seen in free solution. Other advantages ranging from ease of use and reduction in sample size to the potential for automation were also noted with the immobilized enzyme cartridge. Normal free solution proteolysis generally requires 12-24 h, based on the lower enzyme-to substrate ratio in solution. A disturbing feature noted in these lengthy free solution reactions was the tendency to form disulfide bridged peptide artifacts. This could lead to the erroneous conclusion that disulfide bonding in a sample was not that of the native protein. It is concluded that the advantage of immobilized enzymes over free-solution reactions will be most important in the pharmaceutical industry where proteolytic fragment "fingerprinting" of recombinant proteins is being used to confirm structure. PMID- 10669292 TI - Natural poly-histidine affinity tag for purification of recombinant proteins on cobalt(II)-carboxymethylaspartate crosslinked agarose. AB - A natural 19-amino-acid poly-histidine affinity tag was cloned at the N-terminus of three recombinant proteins. The vectors containing the DNA of the fusion proteins were used for transformation of Escherichia coli DH5alpha cells. Each protein was expressed, extracted and purified in one chromatographic step. The purification procedure for each protein can be accomplished in less than 1 h. A new type of immobilized metal ion affinity chromatography adsorbent--Co2+ carboxymethylaspartate agarose Superflow--was utilized at linear flow-rates as high as 5 cm/min. The final preparation of each protein is with purity greater than 95% as ascertained by sodium dodecyl sulfate-electrophoresis. Recovery for each purified protein was higher than 77% of the initial loaded amount as judged by biological activity. The operational capacity of Co2+-carboxymethylaspartate agarose for each protein was determined. PMID- 10669293 TI - A reversed-phase high-performance liquid chromatographic method to analyze retinal isomers. AB - A high-performance liquid chromatographic (HPLC) procedure was developed to separate all-trans-, 13-cis-, 11-cis- and 9-cis-retinal isomers. Two reversed phase Vydac C18 columns in series were used with an isocratic solvent system of 0.1 M ammonium acetate-acetonitrile (40:60, v/v) as mobile phase and all-trans-9 (4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8-no natetraene-1-ol (TMMP) as internal standard. Prior to HPLC, the retinal isomers were efficiently extracted in their original isomeric conformation using dichloromethane-n-hexane in the presence of formaldehyde. This technique is suitable for the assay of 11 cis- and all-trans-retinal isomers in retina. PMID- 10669294 TI - Ion chromatography determination of trace level bromate by large volume injection with conductivity and spectrophotometric detection after post column derivatisation. AB - Bromate is a well known by-product produced by the ozonisation of drinking water; the allowed concentration for human consumption has to be regulated to the low microg l(-1) range. A direct injection, ion chromatographic method was developed using a tetraborate eluent with serially connected conductivity and spectrophotometric detection. Bromate was detected after post-column reaction with fuchsin at 520 nm. Sample capacity was investigated by injecting large volumes (up to 6 ml) using a high total hardness and chloride tap water. Linear correlation of bromate response with volumes from 1 ml to 6 ml was demonstrated, the main limitation being the overlapping of the chloride peak with bromate. Up to 1.5 ml sample can be injected without any pre-treatment. With more than 1.5 ml injection volume, a sample pre-treatment with a cartridge in Ag and H form, followed by a 10 min degassing in an ultrasonic bath, was needed. This method was validated by analysing secondary reference materials and real samples from a drinking water treatment plant. The method was linear from the limit of quantification to 20 microg l(-1). Reproducibilities in tap water were 18% (5 microg l(-1), n=12) and 21% (1 microg l(-1), n=4) respectively for 1.5 and 6 ml injection volumes with conductivity detection, and 17% at 0.5 microg l(-1) (n=9) with spectrophotometric detection. Calculated detection limits were 0.5 microg l( 1) (6 ml) ahd 2 microg l(-1) (1.5 ml) for conductivity detection and 0.3 microg l(-1) (1.5 ml) for spectrophotometric detection. PMID- 10669295 TI - Chiral high-performance liquid chromatography of N-octyl bicycloheptene dicarboximide and confirmatory studies using liquid chromatography-tandem mass spectrometry and two-dimensional nuclear magnetic resonance spectroscopy. AB - N-Octyl bicycloheptene dicarboximide (MGK 264) has exo and endo diastereomers. Each structure has a chiral center at the nitrogen side chain. Enantioselective separation of MGK 264 was achieved by normal-phase high-performance liquid chromatography (HPLC) using cellulose-based Chiralcel OD column with diode-array and optical rotation detectors. Peaks were isolated with the purpose of identifying their stereochemical structures. Molecular mass of the HPLC peaks and their structural information was determined by liquid chromatography-electrospray tandem mass spectrometry (LC-ES-MS-MS). A two-dimensional nuclear magnetic resonance (NMR) spectroscopic technique was used to establish the structural features. Correlation of the data obtained from chiral separation and NMR facilitated in unambiguous assignment of the HPLC peaks. PMID- 10669296 TI - Characterisation of hydrolysable tannins from leaves of Betula pubescens by high performance liquid chromatography-mass spectrometry. AB - A high-performance liquid chromatography-electrospray ionisation mass spectrometry (HPLC-ESI-MS) method, assisted by diode array detection, for the characterisation of individual hydrolysable tannins in birch leaves was developed. With the method, it was found that birch (Betula pubescens) leaves contained an exceptionally complex mixture of hydrolysable tannins; 14 gallotannins and 20 ellagitannins were identified. The developed HPLC-ESI-MS method allows the qualitative and quantitative determination of individual gallotannins and ellagitannins directly from crude birch leaf extract. This is important in studying ecological functions of these phenolic compounds, especially their role in the resistance of birch leaves against insects. PMID- 10669297 TI - Heats of adsorption of some organic compounds on beta-cyclodextrin determined by gas-solid chromatography. AB - Isosteric adsorptive enthalpies have been derived from the temperature dependence of retention volumes determined by eluted pulse gas-solid chromatography. The heat data were obtained for systems using more than 20 organic liquids as adsorbates, and beta-cyclodextrin as adsorbent. The experimental results have been discussed in the light of intermolecular force between molecules of adsorbate and adsorbent. PMID- 10669298 TI - Application of gas chromatography-cryocondensation-Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry to the identification of gas phase reaction products from the alpha-pinene/ozone reaction. AB - The gas phase reaction of alpha-pinene with the atmospheric oxidant ozone was investigated by using the capabilities of both gas chromatography cryocondensation-Fourier transform infrared spectroscopy (GC-FT-IR) and gas chromatography-mass spectrometry (GC-MS), for the identification of the reaction products formed. The reaction was carried out in a flow reaction chamber from where the compounds were sampled on Tenax-containing adsorption cartridges. The reaction mixture was injected onto the column after thermodesorption and analyzed using both GC-IR and GC-MS. Twenty compounds could be detected, including the reactant alpha-pinene and it's impurities tricyclene and camphene. Eleven compounds were identified by spectra comparison with either reference data or spectra obtained from commercial standards. Four compounds were tentatively identified from their IR and MS spectra, while from the remaining two compounds the nature of basic functional groups could be established. PMID- 10669299 TI - Simultaneous determination of phenolic acids and 2,4-dihydroxy-7-methoxy-1,4 benzoxazin-3-one in wheat (Triticum aestivum L.) by gas chromatography-tandem mass spectrometry. AB - A procedure using gas chromatography and tandem mass spectrometry (GC-MS-MS) has been developed for the identification and quantification of some allelochemicals in wheat (Triticum aestivum L.). The quantities of allelochemicals in wheat shoots ranged from 2.9 to 110 mg per kilogram of dry shoot residues. Compared with gas chromatography-mass spectrometry (GC-MS), the GC-MS-MS technique significantly increased instrument selectivity and sensitivity, thereby providing more reliable quantitation results in the determination of the phytotoxic compounds examined during this allelopathy research. PMID- 10669300 TI - Nonaqueous capillary electrophoresis equipped with amperometric detection for analysis of chlorinated phenolic compounds. AB - Nonaqueous capillary electrophoresis (NACE) equipped with amperometric detection has been developed for separation and detection of an 11-member model mixture of chlorinated phenolic compounds. With triacetyl-beta-cyclodextrin (TACD) as a novel selectivity selector, acetonitrile proved to be an excellent solvent for this water-insoluble cyclodextrin derivative. Resolution of the analytes was achieved by using an optimized acetonitrile medium consisting of 500 mM acetic acid, 10 mM sodium acetate, 12 mM TACD and 50 mM tetrabutylammonium perchlorate. Separation of analytes was attributed to differential electrostatic and/or inductive interactions of the analytes with the TACD/TBA+ complex and charged tetrabutylammonium phases. A simple end-column amperometric detector (Pt vs. Ag/AgCl, poised at +1.6 V) in conjunction with NACE was used to analyze chlorophenols. Amperometric detection of such target compounds in acetonitrile based media offers high sensitivity and alleviates electrode fouling compared to aqueous buffers. The detection limits obtained, ranging from 30 nM to 500 nM, are 3-8-fold lower than those obtained with aqueous buffers. PMID- 10669301 TI - Capillary zone electrophoretic analysis of positively charged poly(ethylene oxide) macromolecules using non-covalent polycation-coated fused-silica capillary and indirect UV detection. AB - Capillary zone electrophoresis was used to show the coupling between NH2 terminated poly(ethylene oxide) and oligomers of lactic acid activated by transforming carboxyl chain ends to acyl chloride ones. The demonstration was based on the use of fused-silica capillary physically modified by pre-adsorption of polycations in the reversed polarity mode. As poly(ethylene oxide) macromolecules are UV transparent, indirect UV detection was used. A creatinine solution at pH 4.8 was selected as background electrolyte. Commercially available polycations with different structures were tested. It was shown that the reversed electroosmosis could be modulated according to the structure of the polycation. The method was then applied to analyse a commercial alpha,omega-diamino poly(ethylene oxide), namely Jeffamine ED 600 characterised by a broad mass dispersion. Data showed that the method can detect and separate amino poly(ethylene oxide) of different structures. When applied to analyse post coupling products, no free NH2-terminated poly(ethylene oxide) segments were detected. Moreover, the method allowed detection of water-soluble oligomers generated by partial degradation of lactic segments during the reaction. PMID- 10669302 TI - On the performance and inertness of different materials used for the enrichment of sulfur compounds from air and gaseous samples. AB - The performance of the sorbent polydimethylsiloxane (PDMS) is compared to that of the adsorbents Carbotrap and Tenax for the enrichment of volatile and reactive sulfur compounds. These included: 1- and 2-propanethiol, tetrahydrothiophene, 2 thioethanol and 2-ethylthioethanol. Several artifact-forming reactions were identified on both Tenax and Carbotrap including: H2S elimination and dimerization of thiols. Additionally, permanent adsorption was also observed for heavier solutes. These effects are absent when PDMS is applied. This superior performance is explained by the absence of catalytic or adsorptive activity on PDMS. PMID- 10669303 TI - New gas chromatographic method for residue determination of dithiopyr in soil, wheat grain and straw. AB - A gas chromatographic method has been developed for a new pyridine herbicide, dithiopyr, utilising an electron capture detector. The method makes use of a column (10 m x 0.534 mm I.D.; 1 microm film thickness) containing HP-1 with nitrogen as a carrier gas at a flow rate of 15 ml min(-1) at temperatures of 190, 210, 270 degrees C for oven, injector port and detector, respectively. Soil, wheat grain and straw samples fortified with dithiopyr were extracted with acetone-0.2 M HCl (95:5) and cleaned up by partitioning with hexane. The technique has a sensitivity of 0.05 microg ml(-1) and the recovery of dithiopyr from soil, wheat grain and straw ranged between 80 and 99%. PMID- 10669304 TI - Lipid chain dynamics in stratum corneum studied by spin label electron paramagnetic resonance. AB - The lipid chain motions in stratum corneum (SC) membranes have been studied through electron paramagnetic resonance (EPR) spectroscopy of stearic acid spin labeled at the 5th, 12th and 16th carbon atom positions of the acyl chain. Lipids have been extracted from SC with a series of chloroform/methanol mixtures, in order to compare the molecular dynamics and the thermotropic behavior in intact SC, lipid-depleted SC (containing covalently bound lipids of the corneocyte envelope) and dispersion of extracted SC lipids. The segmental motion of 5- and 12-doxylstearic acid (5- and 12-DSA) and the rotational correlation time of 16 doxylstearic acid (16-DSA) showed that the envelope lipids are more rigid and the extracted lipids are more fluid than the lipids of the intact SC over the range of temperature measured. The lower fluidity observed for the corneocyte envelope, that may be caused mainly due to lipid-protein interactions, suggests a major contribution of this lipid domain to the barrier function of SC. Changes in the activation energy for reorientational diffusion of the 16-DSA spin label showed apparent phase transitions around 54 degrees C, for the three SC samples. Some lipid reorganization may occur in SC above 54 degrees C, in agreement with results reported from studies with several other techniques. This reorganization is sensitive to the presence of the extractable intercellular lipids, being different in the lipid-depleted sample as compared to native SC and lipid dispersion. The results contribute to the understanding of alkyl chain packing and mobility in the SC membranes, which are involved in the mechanisms that control the permeability of different compounds through skin, suggesting an important involvement of the envelope in the skin barrier. PMID- 10669305 TI - Detection of lipid domains in docasahexaenoic acid-rich bilayers by acyl chain specific FRET probes. AB - A major problem in defining biological membrane structure is deducing the nature and even existence of lipid microdomains. Lipid microdomains have been defined operationally as heterogeneities in the behavior of fluorescent membrane probes, particularly the fluorescence resonance energy transfer (FRET) probes 7-nitrobenz 2-oxa-1,3-diazol-4-yl-diacyl-sn-glycero-3-phosphoethan olamine (N-NBD-PE) and (N lissamine rhodamine B sulfonyl)-diacyl-snglycero-3-phosphoethanolamine (N-Rh-PE). Here we test a variety of N-NBD-PEs and N-Rh-PEs containing: (a) undefined acyl chains, (b) liquid crystalline- and gel-state acyl chains, and (c) defined acyl chains matching those of phase separated membrane lipids. The phospholipid bilayer systems employed represent a liquid crystalline/gel phase separation and a cholesterol-driven fluid/fluid phase separation; phase separation is confirmed by differential scanning calorimetry. We tested the hypothesis that acyl chain affinities may dictate the phase into which N-NBD-PE and N-Rh-PE FRET probes partition. While these FRET probes were largely successful at tracking liquid crystalline/gel phase separations, they were less useful in following fluid/fluid separations and appeared to preferentially partition into the liquid-disordered phase. Additionally, partition measurements indicate that the rhodamine containing probes are substantially less hydrophobic than the analogous NBD probes. These experiments indicate that acyl chain affinities may not be sufficient to employ acyl chain-specific N-NBD-PE/N-Rh-PE FRET probes to investigate phase separations into biologically relevant fluid/fluid lipid microdomains. PMID- 10669306 TI - Chemical degradation of liposomes by serum components detected by NMR. AB - Interaction between serum components and liposomes is an oxygen-dependent exothermic process. We studied the interaction of 100 nm extruded liposomes (bearing positive, negative or no charge) with foetal calf serum by 1H NMR and 13C NMR, in order to further our understanding of these reactions. Studies of aqueous or organic extracts obtained after 2 h, 1 day or 1 week, showed hydrolysis to be a degradation process concomitant with the interaction with serum. Oxidation was identified as additional to hydrolysis in the process of degradation. Oxidation produced aldehydes, acids and alcohols, although aldehydes and alcohols were prone to further decomposition and only appeared transiently. Alkenes and other oxidized compounds predominated in those products derived from oxidation. In stearylamine-containing liposomes some aldehydes and a nitroderivative were found as degradation products. Such metabolites are apolar and their presence might explain the intrinsic toxicity of this kind of liposome in cell cultures. The work described in the present study revealed the chemical degradation of liposomes in the serum used. In all cases the results obtained were compared with liposomes not incubated with serum. PMID- 10669307 TI - Interaction of hyaluronic acid-linked phosphatidylethanolamine (HyPE) with LDL and its effect on the susceptibility of LDL lipids to oxidation. AB - The amphiphilic polysaccharide hyaluronic acid-linked phosphatidylethanolamine (HyPE), synthesized by covalently binding dipalmitoyl-phosphatidylethanolamine (DPPE) to short chain hyaluronic acid (mol. wt. approximately = 30 000), interacts with low-density lipoproteins (LDL), to form a 'sugar-decoration' of the LDL surface. This results in an increase in the apparent size of the LDL particles, as studied by photon correlation spectroscopy, and in broadening of the 1H NMR signals of the LDL's phospholipids. Experiments conducted with fluorescently-labeled HyPE indicate that the interaction of HyPE with LDL involves incorporation of the hydrocarbon chains of this amphiphilic polysaccharide into the outer monolayer of the LDL. This interaction also inhibits the copper-induced oxidation of the LDL polyunsaturated fatty acids, avoiding oxidation altogether when the concentration of HyPE is higher than a tenth of the concentration of the LDL's phospholipids. This can not be attributed to competitive binding of copper by HyPE. We propose that the protection of LDL lipids against copper-induced oxidation is due to formation of a sugar network around the LDL. PMID- 10669308 TI - Apolipoprotein A-I localization and dipalmitoylphosphatidylcholine dynamics in reconstituted high density lipoproteins. AB - The structure and molecular dynamics of recombinant high density lipoproteins (rHDL) were studied by non-radiative energy transfer (NRET), fluorescence anisotropy and intensity measurements. The rHDL particles contained human plasma apolipoprotein (apo) A-I and dipalmitoylphosphatidylcholine (DPPC). Fluorescent cis- and trans-parinaric acids were used both as probes of molecular motion in the particle lipid phase and as acceptors in the Forster's energy transfer from apo A-I tryptophan residues to determine particle dimensions, apolipoprotein localization and lipid dynamics. The probes are sensitive to thermal wobbling (macromobility) and conformational deformations (micromobility) of phospholipid acyl chains. The experimental data fitted to various models of the particle structure are compatible with the following: (a) at T < Tt the particles appeared as lens-like discs with a radius of the lipid phase of 5 nm and a mean thickness of 4 nm, the value being more by 20% in the particle centre, the alpha-helices of about 1 nm thickness were located around the edge of the lipid core. Compared to liposomes, both macro- and micromobility of DPPC molecules in rHDL were more rapid due to a significant disorder of the boundary lipid molecules close to the apo A-I molecule. This disorder led to the increase of the specific surface area per one lipid molecule, S(o). The lipid phase can be divided into three regions: (i) zone I of the most tightly packed lipid (0-1.7 nm from the disc axis) with a S(o) value small as 0.5 nm2; (ii) intermediate zone II (from 1.7 to 4.0 nm); and (iii) boundary lipid zone III (4-5 nm) of significantly disordered lipid with a S(o) value large as 0.65 nm2. (b) at T> Tt the S(o) heterogeneity disappeared, the radius of the lipid phase did not increase significantly, not exceeding 5.2 5.4 nm, but protein-induced immobilization of lipid molecules which affected about half or more of the total lipid, became remarkable. The overall effect was the suppression of the transition amplitude in rHDL compared to liposomes. The structural inhomogeneity might underlie the function of the native plasma HDL as the key component of the transport and metabolism of plasma lipids. PMID- 10669309 TI - Two-enzyme system for the synthesis for 1-lauroyl-rac-glycerophosphate (lysophosphatidic acid) and 1-lauroyl-dihydroxyacetonephosphate. AB - A combination of two enzymes, phospholipase D (PL D) and C (PL C), was investigated for the production of two lysophospholipids, 1-lauroyl-rac glycerophosphate (1-LGP) and 1-lauroyl-dihydroxyacetonephosphate (1-LDHAP). The high transphosphatidylation ability of phospholipase D from Streptomyces sp. allowed the formation of 1-lauroyl-phosphatidylglycerol (1-LPG) and 1-lauroyl phosphatidyldihydroxyacetone (1-LPDHA) from phosphatidylcholine (PC) and 1 monolauroyl-rac-glycerol (1-MLG) and 1-lauroyl-dihydroxyacetone (1-MDHA), respectively. A two-phase system, diethyl ether/water, was chosen for the convenience of the recovery of the water insoluble products. A similar two-phase system was used for hydrolysis of the complex phospholipids by phospholipase C form Bacillus cereus, which released both lysophospholipids. Only trace amounts of phosphatidic acid (PA) were detected showing that the enzyme is highly selective for the release of the diacylglycerol and 1-lauroyl-rac glycerophosphate and 1-lauroyl-dihydroxyacetonephosphate. PMID- 10669310 TI - A sensitive endocannabinoid assay. The simultaneous analysis of N acylethanolamines and 2-monoacylglycerols. AB - Mammalian cells produce both N-arachidonoylethanolamine (20:4n-6 NAE, anandamide) and 2-arachidonoylglycerol (2-AG), lipid signaling molecules that activate cannabinoid receptors. Because both agonists occur in the presence of receptor inactive congeners, we have developed a sensitive method for the simultaneous assay of N-acylethanolamines (NAEs) and 2-monoacylglycerols (2-MAG). These lipid classes are isolated from total lipids by solid phase extraction and converted to tert-butyldimethylsilyl (tBDMS) derivatives in the presence of deuterated analogs. The tBDMS derivatives are analyzed by gas chromatography/mass spectrometry using selected ion monitoring programs specific for NAE and 2-MAG. Individual NAEs and 2-MAGs can be quantified in the nanogram and subnanogram range. The NAE and 2-MAG compositions of rat organs and cultured JB6 cells are reported. PMID- 10669311 TI - Cholesterol-induced alterations of the packing properties of gangliosides: an EPR study. AB - The effect of cholesterol (Chol) on two kinds of glycolipid assemblies, one composed of monosialogangliosides (GM1a) and the other formed by a natural mixture of bovine brain gangliosides (TBG), has been analysed. The experimental approach involves spin label electron paramagnetic resonance (EPR) in aqueous lipid dispersions. The employment of a hydrosoluble spin label and a 'quencher' of the EPR signal that is not able to permeate lipid interfaces, allowed us to conclude that GM1a/Chol mixtures give rise to vesicles at Chol proportions for which TBG/Chol mixtures form micelles. The use of different liposoluble spin labels reveals that cholesterol produces a straightening of the hydrocarbon chains in both lipid systems. In GM1a/Chol mixtures, this feature is more pronounced and it is coupled with a decrease in polarity at the chain ends. PMID- 10669312 TI - The structure and function of gramicidin A embedded in interdigitated bilayer. AB - The effects of phase transition from normal to interdigitated lipid bilayer on the function and structure of membrane proteins were studied using linear gramicidin (gramicidin A) as a model. Interdigitated bilayer structure of dipalmitoylphosphatidylglycerol (DPPG) liposomes that was induced by atropine could not be changed notably by intercalating of gramicidin. The K+ transportation of gramicidin in both normal and interdigitated bilayer was assayed by measuring the membrane potential. Results showed that gramicidin in interdigitated bilayer exhibited lower transport capability. Intrinsic fluorescence spectrum of gramicidin in interdigitated bilayer blue-shifted 2.8 nm from the spectrum in normal bilayer, which means that interdigitation provides a more hydrophobic environment for gramicidin. Circular dichroism measurement results indicated that the conformation of gramicidin in interdigitated bilayer is not the typical beta6.3 helix as in the normal bilayer. The results suggested that the interdigitated lipid bilayer might largely affect the structure and function of membrane proteins. PMID- 10669313 TI - Thermal destabilization of transmembrane proteins by local anaesthetics. AB - Local anaesthetics, in addition to anaesthesia, induce the synthesis of heat shock proteins (HSPs), sensitize cells to hyperthermia, and increase the aggregation of nuclear proteins during heat shock. Anaesthetics are membrane active agents, and anaesthesia appears to be due to altered ion channel activity; however, the direct effect of heat shock is protein denaturation. These observations suggest that local anaesthetics may sensitize cells to hyperthermia by interacting with and destabilizing membrane proteins such that protein denaturation is increased. It is shown, using differential scanning calorimetry (DSC), that the local anaesthetics procaine, lidocaine, tetracaine and dibucaine destabilize the transmembrane domains of the Ca2+ -ATPase of sarcoplasmic reticulum and the band III anion transporter of red blood cells. The transmembrane domain of the Ca2+ -ATPase has a transition temperature (Tm) of denaturation of 61 degrees C which is decreased, for example, to 53 degrees C by 15 mM lidocaine. The degree of destabilization (deltaTm) by each anaesthetic is proportional to the lipid to water partition coefficient, and the increased sensitization by anaesthetics with larger partition coefficients and at higher pH suggests that the uncharged forms of the anaesthetics are responsible for destabilization. A Hill analysis of deltaTm for the Ca2+ -ATPase as a function of the concentration of anaesthetic in water gives dissociation constants (Kd) on the order of 10(-4) M, if binding occurs directly from the aqueous phase. This demonstrates moderate affinity binding. However, dissociation constants of 1-3 M are obtained, if binding occurs through the lipid phase, which demonstrates low affinity binding. Thus, the interaction of local anaesthetics with the Ca2+ ATPase may be moderately specific or non-specific depending on the mechanism of interaction. The observation that local anaesthetics also destabilize the transmembrane domain of the band III protein of erythrocytes suggests that destabilization of transmembrane proteins is a general property of anaesthetics, which is at least in part a mechanism of sensitization to hyperthermia. PMID- 10669314 TI - Apoptosis induced by hyperthermia in Dunn osteosarcoma cell line in vitro. AB - The effect of hyperthermia at 43.5 degrees C for 1 h on Dunn osteosarcoma cells was studied. With sham-heated cells (37 degrees C, 1 h) as the control, the hyperthermia treated cells were divided into five groups. Time 0 group was the cells that were harvested immediately after heated at 43.5 degrees C for 1 h. Whereas time 3, 6, 12, and 24 h groups were the cells that were collected respectively after reincubation at 37 degrees C for the above different time periods. The appearance of hyperthermia-induced apoptosis of Dunn osteosarcoma cells was demonstrated to be time dependent. With the confocal microscopic study and TUNEL staining, the morphological characteristics of apoptosis, condensed nuclei and fragmented nuclei were obvious when reincubated at 37 degrees C for 6 h after hyperthermic treatment. This hyperthermia-induced apoptosis was further confirmed by flow cytometric analysis on DNA contents. The sub-G1 region that was proposed as a marker of apoptotic cells was most significantly elevated at 6 h after hyperthermic treatment and, thereafter, decreased to the levels of control values by 24 h, as the apoptotic cells underwent secondary necrosis and degraded to debris. The DNA strand breaks, considered as the key biochemical event of apoptosis, were detected by the TUNEL assay. This study indicated that hyperthermia (43.5 degrees C for 1 h) can induce apoptotic changes on osteosarcoma cells in vitro very rapidly (within 6 h after treatment), and its occurrence might not be detected if the samples are not taken at several early time points after hyperthermia. PMID- 10669315 TI - Comparison of the effects of two different whole body hyperthermia protocols on the distribution of murine leukocyte populations. AB - Two predominant WBH protocols presently being used in clinical trials include a low temperature, long duration (LL) WBH, where core body temperature is raised to 39.5-40 degrees C for 6h or more, and a high temperature, short duration (HS) WBH, where core body temperature is raised to 41.8 degrees C for up to 2h. Here, the effects of LL-WBH and HS-WBH on leukocyte populations in the blood, spleen, lymph node (LN) and peritoneal cavity (PerC) of Balb/c mice were compared using flow cytometry. The total numbers of peripheral blood leukocytes decreased up to 2-fold immediately after LL-WBH, reflecting a decrease of lymphocyte numbers compared to controls. In contrast, the numbers of blood leukocytes are increased 2.7-fold immediately after HS-WBH compared to controls, reflecting an increase in lymphocytes, monocytes and granulocytes. After both LL- and HS-WBH treatment, leukocyte numbers in the spleen are decreased approximately 2-fold, again reflecting decreases in lymphocyte numbers. In the PerC, total numbers of leukocytes are also significantly decreased (2-fold) during LL-WBH but not HS WBH. Total numbers of leukocytes in the LNs were unaffected by both LL- and HS WBH. Overall, these data reveal differential effects of the LL- and HS-WBH protocols on leukocyte populations in the blood, spleen, LN and PerC of Balb/c mice. PMID- 10669316 TI - Fever-range hyperthermia stimulates alpha4beta7 integrin-dependent lymphocyte endothelial adhesion. AB - Migration of blood-borne lymphocytes into lymphoid tissues is initiated by the L selectin and alpha4beta7 integrin adhesion molecules. Previous studies have shown that L-selectin adhesion is dynamically regulated by febrile temperatures. It is now reported that fever-range hyperthermia also acts directly on lymphocytes to enhance selected adhesive functions of alpha4beta7 integrin. Fever-range hyperthermia treatment in vitro (40 degrees C, 12 h) of murine TK1 lymphoma cells and human peripheral blood lymphocytes (PBL) stimulates alpha4beta7 integrin dependent adhesion to high endothelial venules (HEV) in Peyer's patch and mesenteric lymph node frozen sections. TK1 cells are alpha4beta7hi L selectin(lo), allowing for the analysis of alpha4beta7 integrin without contributions from L-selectin. Adhesion was further shown to involve alpha4beta7 integrin and its endothelial counter-receptor, mucosal addressin cell adhesion molecule-1 (MAdCAM-1) using function-blocking antibodies (i.e. DATK32, HP2/1, MECA-367). Fever-range hyperthermia also promotes alpha4beta7 integrin-mediated aggregation of TK1 cells. In sharp contrast, hyperthermia fails to increase alpha4beta7 integrin adhesion to fibronectin by TK1 cells. Expression of the alpha4beta7 heterodimer on TK1 cells or human PBL is not altered by hyperthermia, suggesting that hyperthermia stimulates adhesion by enhancing alpha4beta7 integrin avidity rather than its cell surface density. These results provide a mechanism whereby febrile temperatures during infection or clinical hyperthermia potentially amplify the immune response by stimulating L-selectin and alpha4beta7 integrin-dependent homing of immune effector cells to lymphoid tissues. PMID- 10669318 TI - Acute histological effects of interstitial hyperthermia on normal rat brain. AB - Histological changes in the brains of Fischer rats at different times after interstitial heating with various thermal doses were studied. The brains, subjected to sham-heating, and heating at 39 and 40 degrees C for 30 min showed mild capillary congestion and minimal vacuolation at 4, 24 and 72 h. In the brains heated to 41, 42 and 43 degrees C for 30 min, there was local vascular congestion, petechiae, vacuolation and cellular shrinkage with nuclear pyknosis at 4 h; enhanced congestion and petechiae, acute cellular necrosis, infiltration of polymorphonuclear leukocytes and marked vacuolation at the margin at 24h; total coagulative necrosis of all parenchymal and vascular elements, early liquefaction necrosis and vascular hyperplasia at the margin at 72 h; enhanced vascular hyperplasia at the margin at 120 h and 168 h. The threshold thermal dose for the histopathological damage in the rat brain was heating at 41 degrees C for 30 min. PMID- 10669317 TI - Hyperthermia and paclitaxel--epirubicin chemotherapy: enhanced cytotoxic effect in a murine mammary adenocarcinoma. AB - Multimodality therapy is considered of great interest in the treatment of locally advanced solid tumours. In previous experiments, paclitaxel (TX) and epirubicin (EP) were combined with different schedules, obtaining a superadditive effect on the growth of a murine mammary carcinoma. In the present study, the authors have analysed the possible use of hyperthermia (HT) to increase the efficacy of TX and EP combinations. Tumours were transplanted into the right hind foot of female hybrid (C3D2F1) mice. Both TX and EP were administered i.p in two different doses. Hyperthermia was applied using a water bath at 43.2 degrees C for 1 h. Results were analysed in terms of Tumour Growth Delay (TGD). The maximum tolerated doses in combined protocols were TX 45 mg/kg and EP 9 mg/kg, with an interval time of 24h between the two administrations. TGDs of some of the schedules performed are reported: EP + HT = 11 days, TX + HT = 16 days, TX + EP (with an interval time of 24 h) = 14 days, and TX + EP + HT = 22 days. In the experimental model, HT significantly increases the effects of both TX and EP. TX + EP + HT treatment is the most effective (significantly different from TX + EP), but not in a significant way when compared to TX + HT treatment. These results suggest the possible use of a TX + HT protocol for local tumour response, whereas EP could be added in order to achieve a better systemic control. PMID- 10669319 TI - Hyperthermia and radiotherapy for inoperable squamous cell carcinoma metastatic to cervical lymph nodes from an unknown primary site. AB - INTRODUCTION: Neck node metastases from an unknown primary carcinoma represent an infrequent but challenging problem for oncologists. The management of such patients is controversial, but radiotherapy alone or as part of a multimodal approach is often indicated. Patients with inoperable lesions usually receive radiotherapy alone at palliative doses. In an attempt to increase local control in patients with locally advanced neck disease from an unknown primary carcinoma, local hyperthermia was combined with definitive radiotherapy. MATERIAL AND METHODS: Between 1982 and 1993, radiotherapy and local microwave hyperthermia were used to treat 15 patients with metastatic neck nodes from an unknown primary site. The patients had previously undergone only biopsy or fine needle biopsy, and showed no signs of metastases beyond the clavicle. Radiation to the nodes and the potentially primary sites in the head and neck was delivered by a 6 MV linear accelerator or a Cobalt 60 unit, to a total dose of 57.50-74.40 Gy (median 70 Gy). Hyperthermia was added using a BSD 1000 unit at an operating frequency of 280-300 MHz for 2-7 sessions (mean 3.1; median 2) at a desired minimum temperature of 42.5 degrees C. Two patients also received i.v. cisplatin 20 mg/m2/week as a radiosensitizer. RESULTS: Nine patients achieved a complete, and four a partial response for an overall response rate of 86.5%. Acute and late toxicity was mild: four patients experienced pain during hyperthermia, two moist cutaneous desquamation, and one cutaneous necrosis. The actuarial probability of maintaining local control at 5 years is 64.5% and the actuarial overall survival 29%. Five patients developed distant metastases and died of disease, two experienced nodal recurrence and two died of other unrelated causes. CONCLUSION: The addition of local microwave hyperthermia to radiotherapy in the treatment of metastatic squamous cell carcinoma of the neck in patients with an unknown primary site leads to good local control with moderate toxicity. No definite conclusions are possible because of the small number of patients involved in this phase II trial. PMID- 10669320 TI - Elimination of new chronic hepatitis B virus infections: results of the Alaska immunization program. AB - An immunization assessment and a serologic survey were conducted to evaluate the effectiveness of a hepatitis B immunization program in eliminating hepatitis B virus (HBV) transmission among Alaska Natives in a region in which HBV infection is endemic. Hepatitis B vaccine coverage was 93% among 567 children 50 copies/mL. HIV DNA decreased in patients with <400 RNA copies/mL at baseline and showed an HIV RNA increase after vaccination (n=8) when compared with 8 patients with <50 copies/mL at baseline who lacked viral response to vaccination. Concurrent decreases in proviral DNA and memory phenotype CD4+ cells in association with increased plasma HIV RNA after vaccination in patients with <400 RNA copies/mL at baseline suggest that in vivo mobilization of the latently infected cell reservoir may occur during potent antiretroviral therapy. PMID- 10669336 TI - Secretory anti-human immunodeficiency virus (HIV) antibodies in colostrum and breast milk are not a major determinant of the protection of early postnatal transmission of HIV. AB - The immune response to human immunodeficiency virus (HIV) type 1 was evaluated in breast milk from HIV-infected African mothers who had transmitted and those who had not transmitted HIV to their children through breast-feeding. The levels, specific activities against gp160 and 2 HIV-derived peptides from gp41 and gp120 (V3 loop), and inhibitory activity toward viral transcytosis in vitro of secretory IgA (S-IgA) and IgG purified from breast milk were investigated in 8 transmitting mothers and 18 nontransmitting mothers. S-IgA and IgG antibodies to gp160 and to peptides were found in all breast milk samples. The specific activities of S-IgA and IgG to gp160 and peptides were similar between transmitting and nontransmitting mothers. No difference of the capacity of S-IgA and IgG to block HIV transcytosis in vitro was found between the 2 groups. These results suggest that humoral mucosal immunity to HIV does not appear as a predominant factor for protection against viral transmission through breast milk. PMID- 10669337 TI - A randomized study of the safety and antiretroviral activity of hydroxyurea combined with didanosine in persons infected with human immunodeficiency virus type 1. American Foundation for AIDS Research Community-Based Clinical Trials Network. AB - This randomized open-label trial of human immunodeficiency virus type 1-infected persons compared safety and efficacy for 38 patients receiving hydroxyurea/didanosine combination therapy with findings in 42 persons given didanosine monotherapy for 12 weeks, followed by 12 weeks of hydroxyurea/didanosine combination therapy for all patients. Week 12 on-treatment group comparisons showed a mean decrease in virus load between hydroxyurea/didanosine versus didanosine groups of -0.93 versus -0.74 log10 copies/mL (P=.20); a higher percentage of the hydroxyurea/didanosine group below the assay's detection limit (500 copies/mL), 29% versus 7% (P=.017); and median change in CD4 cells for the hydroxyurea/didanosine versus didanosine group of 0 versus 43 cells/mm3 (P=.045), although median change in CD4 percentage was similar (0.9% vs. 1.2%, P=.64). Week 24 virus load reductions and CD4 cell changes were similar in both groups. Intent-to-treat and on-treatment analyses showed similar results. The hydroxyurea/didanosine combination was well tolerated. PMID- 10669338 TI - Use of human immunodeficiency virus (HIV) human hyperimmune immunoglobulin in HIV type 1-infected children (Pediatric AIDS clinical trials group protocol 273). AB - The clinical, immunologic, and virologic effects and the pharmacokinetics of human immunodeficiency virus (HIV) human hyperimmune immunoglobulin (HIVIG) were assessed in 30 HIV-infected children aged 2-11 years. All had moderately advanced disease with an immune complex-dissociated (ICD) p24 antigen >70 pg/mL and were on stable antiviral therapy. Three groups of 10 children received 6 monthly infusions of 200, 400, or 800 mg/kg of HIVIG, and serial immunologic and virologic assays were performed. HIVIG doses as high as 800 mg/kg were safe and well tolerated. The half-life of HIVIG, determined by serial p24 antibody titers, was 13-16 days, the volume of distribution was 102-113 mL/kg, and clearance was 5.6-6.0 mL/kg/day. Plasma ICD p24 decreased during the infusions, but CD4 cell levels, plasma RNA copy number, cellular virus, immunoglobulin levels, and neutralizing antibody titers were minimally affected by the infusions. Clinical status did not change during the 6-month infusion and 3-month follow-up periods. PMID- 10669339 TI - Genital tract human immunodeficiency virus type 1 (HIV-1) shedding and inflammation and HIV-1 env diversity in perinatal HIV-1 transmission. AB - This study sought to identify genital tract characteristics associated with vertical transmission of human immunodeficiency virus type 1 (HIV-1). HIV-1 DNA and RNA, HIV-1 env diversity, and inflammatory cells were quantified in cervicovaginal lavages (CVLs) of 24 women enrolled in the Women and Infants Transmission Study; 7 women transmitted HIV-1 perinatally. Vaginal candidiasis, HIV-1 culture positivity, levels of HIV-1 DNA and cell-free RNA, and HIV-1 env diversity were significantly higher in the CVLs of transmitters. CVL HIV-1 DNA levels correlated with higher levels of inflammatory cells and cell-free HIV-1 RNA. Of subjects with paired blood and CVL specimens, there was more HIV-1 env heterogeneity between blood and CVLs in transmitters than in nontransmitters. In summary, increased HIV-1 shedding is correlated with a more complex population of HIV-1 quasispecies in the genital tracts of parturient women, which may increase the probability that a fetotropic strain is transmitted. PMID- 10669340 TI - T cells overexpressing interferon-gamma and interleukin-10 are found in both the thymus and secondary lymphoid tissues of feline immunodeficiency virus-infected cats. AB - Similar to human immunodeficiency virus type 1, feline immunodeficiency virus (FIV) replicates in the thymus of infected animals, causing marked alteration in thymic lymphocyte subpopulations. The immune phenotype and cytokine patterns in the thymus and secondary lymphoid tissues of FIV-infected cats were investigated. FIV infection caused an acute-stage transient reduction in CD4CD8 double-positive thymocytes, a marked increase in CD8 single-positive thymocytes, and formation of thymic B cell lymphoid follicles. Interferon (IFN)-gamma and interleukin (IL)-10 mRNA were up-regulated in both the thymus and lymph nodes of FIV-infected cats. Analysis of purified CD4 and CD8 cells revealed that CD4 cells produced most of the IL-10, whereas IFN-gamma was produced by both subsets. Quantitative competitive reverse-transcription polymerase chain reaction analysis revealed that thymocytes, especially CD4CD8 thymocytes, had much greater levels of gag mRNA than did lymph node T cells. Thus, overexpression of IFN-gamma and IL-10 is a feature of the thymus and secondary lymphoid tissues of FIV-infected cats. PMID- 10669342 TI - Intravaginal practices, vaginal flora disturbances, and acquisition of sexually transmitted diseases in Zimbabwean women. AB - One hundred sixty-nine Zimbabwean women were studied to determine whether the use of intravaginal practices (cleaning with the fingers, wiping the vagina, and inserting traditional substances) are associated with disturbances of vaginal flora and acquisition of sexually transmitted diseases (STDs). Subjects were interviewed and received counseling and a pelvic examination at enrollment, 1 month, and 6 months, and vaginal specimens were collected at enrollment and at 6 months. Users were more likely than nonusers to have vaginal flora disturbances but were not more likely to acquire an STD (relative risk [RR], 2.15; P=.188). Certain vaginal flora disturbances were associated with increased STD incidence and HIV prevalence. The absence of lactobacilli from the vaginal flora was associated with being positive for human immunodeficiency virus in baseline (odds ratio [OR], 0.24; P=.001) and 6-month transition multivariate models (OR, 0.39; P=.025). The presence of clue cells at baseline was associated with a higher incidence of STDs (RR, 1. 94; P=.025). PMID- 10669341 TI - Exogenous glucocorticoids alter parameters of early feline immunodeficiency virus infection. AB - Feline immunodeficiency virus (FIV), a lentivirus, causes progressive immunosuppression and neurologic dysfunction in cats. Glucocorticoids are common therapeutic agents that are also immunosuppressive, and their use might enhance the pathogenic effects of lentivirus infections. Methylprednisolone acetate, a long-acting glucocorticoid, was administered to cats before FIV inoculation, and the course of early infection was monitored. The humoral immune response to FIV was not affected by corticosteroid treatment, but CD8+ cell-mediated antiviral activity was poor in cultures from FIV-infected cats treated with methylprednisolone. Steroid-treated cats had higher plasma viral RNA levels than untreated cats during acute viremia. In contrast, FIV-associated changes in brain stem auditory-evoked potentials were slow to develop in the methylprednisolone treated cats. Methylprednisolone treatment of cats with established FIV infections appeared to reverse these neurophysiologic changes. These results emphasize the complexity of host-lentivirus interactions and suggest potential advantages and drawbacks of using glucocorticoids in lentivirus infections. PMID- 10669343 TI - Effects of contraceptive method on the vaginal microbial flora: a prospective evaluation. AB - A prospective evaluation of 331 university women who were initiating use of oral contraceptive pills (OCPs), a cervical cap, diaphragm-spermicide, or other spermicidal methods was done to assess the effects of commonly used contraceptive methods on the vaginal flora. Vaginal introital cultures were performed at baseline and then weekly for 1 month. The prevalence of Escherichia coli vaginal colonization and of abnormal vaginal Gram stain scores (Nugent criteria) increased significantly among women using a cervical cap or diaphragm-spermicide but not among women using OCPs. Women with E. coli colonization were significantly more likely to have an abnormal Nugent score and an absence of lactobacilli. In a multivariate model, use of spermicidal contraception in the preceding week was associated with an abnormal Nugent score and with colonization with E. coli, Enterococcus species, and anaerobic gram-negative rods. Thus, spermicidal methods of contraception are associated with alterations of the vaginal microflora that consequently may predispose women to genitourinary infections. PMID- 10669344 TI - Expression of and cytokine activation by Escherichia coli curli fibers in human sepsis. AB - Curli organelles are expressed by commensal Escherichia coli K12 and by Salmonella typhimurium at temperatures <37 degrees C, which bind serum proteins and activate the contact-phase system in vitro. This study demonstrates, by means of an anti-CsgA (curli major subunit) antibody, that a significant fraction of E. coli isolates (24 of 46) from human blood cultures produce curli at 37 degrees C in vitro. Serum samples from 12 convalescent patients with sepsis, but not serum from healthy controls, contained antibodies against CsgA (n=12). This study further demonstrates that a curli-expressing E. coli strain and a noncurliated mutant secreting soluble CsgA induce significantly (P<.05) higher levels of proinflammatory cytokines (tumor necrosis factor-alpha, interleukin [IL]-6, and IL-8) in human macrophages differentiated from THP-1 cells. These data, therefore, provide direct evidence that curli are expressed in vivo in human sepsis and suggest a possible role for curli and CsgA in the induction of proinflammatory cytokines during E. coli sepsis. PMID- 10669345 TI - Interleukin 10 inhibits the release of CC chemokines during human endotoxemia. AB - Sixteen healthy subjects were intravenously injected with lipopolysaccharide (LPS), once with placebo and once with recombinant human interleukin (IL)-10 (25 microgram/kg), to determine the effect of IL-10 on LPS-induced production of macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and monocyte chemoattractant protein (MCP)-1. LPS induced transient increases in serum MIP 1alpha, MIP-1beta, and MCP-1. Pretreatment with IL-10 inhibited LPS-induced release of MIP-1alpha, MIP-1beta, and MCP-1. In whole blood in vitro, the IL-10 induced inhibition of MIP-1alpha and MIP-1beta release was equally potent in the presence or absence of an anti-tumor necrosis factor (TNF) antibody. Although isolated peripheral blood mononuclear cells produced more MIP-1alpha and MIP 1beta than neutrophils, the latter cells were more sensitive to the inhibiting effect of IL-10. IL-10 attenuates LPS-induced production of CC chemokines in human endotoxemia, whereby in vitro experiments suggest that, in the case of MIP 1alpha and MIP-1beta release, this effect is independent from an inhibitory effect on TNF production. PMID- 10669346 TI - Prognostic value of cytokine concentrations (tumor necrosis factor-alpha, interleukin-6, and interleukin-10) and clinical parameters in severe melioidosis. AB - Raised serum concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, or IL-10 are associated with mortality in patients with sepsis, but it is not known whether elevated cytokine levels are independently predictive of mortality. Cytokine assays (TNF-alpha, IL-6, and IL-10) were performed on admission plasma samples from 172 adult Thai patients with severe melioidosis. Mortality was 31.4%. APACHE II score; septicemia; plasma lactate; TNF-alpha, IL 6, and IL-10 concentrations; and IL-10/TNF-alpha and IL-6/IL-10 ratios were each associated with outcome (P>IgG2a=IgG2b=IgG3) in newborn pups closely resembled that in their mothers. Seven of 9 female baboons given the III-TT vaccine had 5- to 36-fold increases in specific antibody from baseline levels; they transferred 26%-185% of specific antibody to their offspring. Matched maternal and neonatal sera obtained at delivery were functionally equivalent in an in vitro opsonophagocytosis assay. These preclinical studies provide further evidence for effective immunogenicity of GBS conjugate vaccine and efficient transport of functionally active maternal antibody. PMID- 10669350 TI - Role of Streptococcus pneumoniae and Haemophilus influenzae in the development of acute otitis media and otitis media with effusion in a gerbil model. AB - The efficacy of amoxicillin/clavulanate and cefuroxime was determined in a gerbil model of otitis media with a mixed Streptococcus pneumoniae plus Haemophilus influenzae middle ear (ME) infection. Results were compared with those obtained in a previous single H. influenzae model. All untreated animals inoculated with the mixed inoculum developed acute otitis media (AOM), whereas 86.7% of those inoculated with H. influenzae developed otitis media with effusion (OME). Antibiotics eradicated H. influenzae from the ME more efficiently in AOM than in OME, and this difference was highly significant (P80% of animals developed culture-negative OME. PMID- 10669352 TI - Analysis of the pathogenicity locus in Clostridium difficile strains. AB - The genes for Clostridium difficile toxins A and B (tcdA and tcdB) are part of a 19.6-kb pathogenicity locus (PaLoc) that includes the genes tcdD, tcdE, and tcdC. To determine whether the C. difficile PaLoc is a stable and conserved genetic unit in toxigenic strains, a multiplex polymerase chain reaction was used to analyze 50 toxigenic, 39 nontoxigenic, and 2 toxin-defective isolates. The respective amplicons were identified for tcdA-E in the toxigenic isolates; these were absent in the nontoxigenic isolates. C. difficile P-829 lacked at least a fragment of tcdD, tcdB, tcdE, and tcdC, but tcdA was present. C. difficile 8864 had deletions in the tcdA and tcdC genes. These data suggest that the PaLoc is highly stable in toxigenic C. difficile, nontoxigenic isolates lack the unit, and isolates with a defective PaLoc can still cause clinical disease. Further studies are needed to define the role of individual genes in the pathogenesis of C. difficile-associated diarrhea. PMID- 10669353 TI - Quinolone antibiotics induce Shiga toxin-encoding bacteriophages, toxin production, and death in mice. AB - Shiga toxin-producing Escherichia coli (STEC) cause significant disease; treatment is supportive and antibiotic use is controversial. Ciprofloxacin but not fosfomycin causes Shiga toxin-encoding bacteriophage induction and enhanced Shiga toxin (Stx) production from E. coli O157:H7 in vitro. The potential clinical relevance of this was examined in mice colonized with E. coli O157:H7 and given either ciprofloxacin or fosfomycin. Both antibiotics caused a reduction in fecal STEC. However, animals treated with ciprofloxacin had a marked increase in free fecal Stx, associated with death in two-thirds of the mice, whereas fosfomycin did not. Experiments that used a kanamycin-marked Stx2 prophage demonstrated that ciprofloxacin, but not fosfomycin, caused enhanced intraintestinal transfer of Stx2 prophage from one E. coli to another. These observations suggest that treatment of human STEC infection with bacteriophage inducing antibiotics, such as fluoroquinolones, may have significant adverse clinical consequences and that fluoroquinolone antibiotics may enhance the movement of virulence factors in vivo. PMID- 10669355 TI - Genetic evidence for the role of the Lv locus in early susceptibility but not IL 10 synthesis in experimental coccidioidomycosis in C57BL mice. AB - Loci on chromosome 4 near Lv and on chromosome 6 near Tnfr1 are associated with resistance to coccidioidomycosis in mice. To assess the importance of the Lv locus, we compared C57BL/6 (B6) with C57BL/10 (B10), strains that are nearly congenic for the Lv locus. Fourteen days after intraperitoneal infection, B6 mice had nearly 100-fold more Coccidioides immitis in their lungs than did B10 mice (log 6.2 vs. log 4.8). Furthermore, the time to 50% deaths was 15 days for B6 and 22 days for B10. Nevertheless, 90% of B10 mice had died by day 28. In other mouse strains, we found a direct correlation between lung colony-forming units and levels of interleukin (IL)-10 and IL-4 mRNA, but B10 mice had 100-fold higher lung levels of IL-10 and 10-fold higher levels of IL-4 mRNA than did B6 mice, despite having less C. immitis. In the absence of IL-10, B10 mice are resistant to lethal infection. These results suggest that a locus near Lv is responsible for early resistance to coccidioidomycosis but not for modulating the IL-10 and IL-4 responses. This locus is not sufficient to make C57BL mice resistant to coccidioidomycosis. PMID- 10669354 TI - Rapid neutrophil response controls fast-replicating intracellular bacteria but not slow-replicating Mycobacterium tuberculosis. AB - Being one of the first cells to invade the site of infection, neutrophils play an important role in the control of various bacterial and viral infections. In the present work, the contribution of neutrophils to the control of infection with different intracellular bacteria was investigated. Mice were treated with the neutrophil-depleting monoclonal antibody RB6-8C5, and the time course of infection in treated and untreated mice was compared by using intracellular bacterial species and strains varying in virulence and replication rate. The results indicate that neutrophils are crucial for the control of fast-replicating intracellular bacteria, whereas early neutrophil effector mechanisms are dispensable for the control of the slow-replicating Mycobacterium tuberculosis. PMID- 10669356 TI - Host immune reactivity determines the efficacy of combination immunotherapy and antifungal chemotherapy in candidiasis. AB - In immunocompetent mice with candidiasis, successful therapy with amphotericin B and fluconazole relies on the induction of protective, T helper (Th) type 1 responses, an effect potentiated by concomitant interleukin (IL)-4 neutralization. To assess the therapeutic efficacy of combined treatments with antifungals and immunomodulators in conditions of immunosuppression, leukopenic or neutropenic mice with disseminated candidiasis were treated with amphotericin B or fluconazole alone or in combination with soluble IL-4 receptor (sIL-4R) or recombinant (r) IL-12 or IL-10 neutralizing monoclonal antibodies. We found that (1) the synergistic effect of sIL-4R and antifungals is retained in immunocompromised mice; (2) synergism with amphotericin B was superior to that with fluconazole, particularly in leukopenic mice; (3) rIL-12 synergized with fluconazole in neutropenic mice; and (4) IL-10 neutralization was always of limited efficacy. This study indicates that the therapeutic efficacy of antifungals is differentially potentiated by cytokines or cytokine antagonists and is influenced by host immune reactivity. PMID- 10669357 TI - An outbreak of cryptosporidiosis linked to a foodhandler. AB - In September and October 1998, a cryptosporidiosis outbreak occurred on a Washington, DC, university campus. In a case-control study of 88 case patients and 67 control subjects, eating in 1 of 2 cafeterias was associated with diarrheal illness (P<.001). Morbidity was associated with eating dinner on 22 September (odds ratio, 8.1; 95% confidence interval, 3.4-19.5); weaker associations were found for 6 other meals. Cryptosporidium parvum was detected in stool specimens of 16 (70%) of 23 ill students and 2 of 4 ill employees. One ill foodhandler with laboratory-confirmed C. parvum prepared raw produce on 20-22 September. All 25 Cryptosporidium isolates submitted for DNA analysis, including 3 from the ill foodhandler, were genotype 1. This outbreak illustrates the potential for cryptosporidiosis to cause foodborne illness. Epidemiologic and molecular evidence indicate that an ill foodhandler was the likely outbreak source. PMID- 10669358 TI - Interferon-gamma expression in jejunal biopsies in experimental human cryptosporidiosis correlates with prior sensitization and control of oocyst excretion. AB - To investigate the role of interferon (IFN)-gamma in human cryptosporidiosis, jejunal biopsies from experimentally infected volunteers and chronically infected AIDS patients were examined for IFN-gamma expression by in situ hybridization. IFN-gamma expression was compared with oocyst excretion, baseline serum anti Cryptosporidium antibody, and symptoms. IFN-gamma mRNA was detected in biopsies from 13 of 26 volunteers after experimental infection but not in biopsies taken before C. parvum exposure or in biopsies from patients with AIDS-associated cryptosporidiosis. After challenge, 9 of 10 volunteers with baseline C. parvum antibody produced IFN-gamma, compared with 4 of 16 volunteers without baseline antibody (P<.01). Furthermore, IFN-gamma mRNA was detected in 9 of 13 volunteers who did not excrete oocysts, compared with 4 of 13 with organisms (P<.05). Thus, expression of IFN-gamma in the jejunum was associated with prior sensitization and absence of oocyst shedding. IFN-gamma production may explain the resistance to infection noted in sensitized persons but may not be involved in control of human primary infection. PMID- 10669359 TI - Continued transmission of West Nile virus to humans in southeastern Romania, 1997 1998. AB - After an epidemic of West Nile (WN) virus neurologic infections in southeastern Romania in 1996, human and animal surveillance were established to monitor continued transmission of the virus. During 1997 and 1998, neurologic infections were diagnosed serologically as WN encephalitis in 12 of 322 patients in 19 southeastern districts and in 1 of 75 Bucharest patients. In addition, amid a countrywide epidemic of measles, the etiology of the febrile exanthem in 2 of 180 investigated cases was determined serologically to be WN fever; 1 case was complicated by hepatitis. Sentinel chickens placed in Bucharest seroconverted to WN virus during the summer months, indicating their potential value in monitoring transmission. The continued occurrence of sporadic WN infections in southeastern Romania in consecutive years after the 1996 epidemic is consistent with local enzootic transmission of the virus. PMID- 10669360 TI - Selection of multiresistant hepatitis B virus during sequential nucleoside analogue therapy. AB - Hepatitis B virus (HBV) drug resistance to lamivudine is always accompanied by mutations in the viral polymerase gene at position 550, termed group 1 (M550V with L526M) or group 2 (M550I) mutations. The latter mutation has not been associated with famciclovir resistance. Thus, the addition of famciclovir to lamivudine therapy in persons with group 2 lamivudine resistance may lead to virus suppression. The effect of lamivudine/famciclovir combination therapy on HBV infection was monitored in 5 lamivudine-resistant patients by quantitative polymerase chain reaction and polymerase gene sequencing of serum virus. No patients treated with combination therapy had a decline in HBV load >1 log10. Continual evolution of the viral polymerase was detected in association with virologic resistance to both drugs. Cloning experiments identified the preexistence of these multidrug-resistant virus variants as minority species prior to addition of famciclovir therapy. HBV resistance to lamivudine monotherapy is associated with a complex mixture of variants that limit the efficacy of second-line nucleoside-analogue therapy. First-line potent combination therapy may reduce the emergence of HBV drug resistance. PMID- 10669361 TI - Cytomegalovirus (CMV) DNA load predicts relapsing CMV infection after solid organ transplantation. AB - Cytomegalovirus (CMV) DNA load was analyzed as a marker for relapse of CMV infection in 24 solid organ transplant patients with CMV infection or disease who received a fixed 14-day course of intravenous ganciclovir. Viral load was measured in blood samples obtained before and at the completion of treatment. Eight (33%) of 24 patients developed relapsing CMV infection. Median pretreatment viral loads were higher in the relapsing group (80,150 copies/106 leukocytes) than in the nonrelapsing group (5500 copies/106 leukocytes; P=.007). The relapsing group also had persistent detectable viral DNA (median, 5810 copies/106 leukocytes) after treatment, whereas it was undetectable in the nonrelapsing group (P<. 0001). Primary CMV infection (seronegative recipients of seropositive organs, D+R-) was an independent marker for CMV relapse (P=.03), and these patients had higher pre- and posttreatment viral loads than did non-D+/R- patients (P<.0001 and P=.0014, respectively). CMV DNA load is a useful marker for individualizing antiviral treatment of CMV infection in solid organ transplant recipients. PMID- 10669362 TI - Uninfected and cytomegalic endothelial cells in blood during cytomegalovirus infection: effect of acute rejection. AB - After transplantation, human cytomegalovirus (HCMV) infections can cause vascular damage to both the graft and the host. To study a possible relationship between the degree of vascular injury, clinical symptoms of HCMV infection, and transplant rejection, the appearance and numbers of endothelial cells (ECs) in blood of 54 kidney transplant recipients were investigated in a prospective clinical study. Two types of endothelial cells were identified: cytomegalic ECs (CECs) were detected in patients with moderate or high HCMV antigenemia, and uninfected ECs were observed in patients with and without HCMV infection. The incidence of either CECs, ECs, or the combination of both was associated with HCMV-related clinical symptoms (P<.01). Remarkably, the occurrence of rejection episodes before HCMV infection was an important risk factor for the occurrence of ECs in blood (ECs, CECs, or both) during HCMV infection (P<.001). PMID- 10669363 TI - Comparison of the safety, vaccine virus shedding, and immunogenicity of influenza virus vaccine, trivalent, types A and B, live cold-adapted, administered to human immunodeficiency virus (HIV)-infected and non-HIV-infected adults. AB - Fifty-seven human immunodeficiency virus (HIV)-infected (CDC class A1-2) and 54 non-HIV-infected adults, not prescreened for influenza susceptibility, were randomized to receive trivalent live attenuated influenza vaccine (LAIV) or placebo intranasally. LAIV was safe and well tolerated with no serious adverse events attributable to vaccine. Reactogenicity rates were similar in LAIV and placebo recipients except that runny nose/nasal congestion was significantly more common in LAIV recipients regardless of HIV status. No prolonged shedding of LAIV was observed in HIV-infected participants. HIV RNA levels were not increased and CD4 counts were not decreased in HIV-infected LAIV recipients compared with placebo recipients after immunization. Shedding of LAIV and increases in antibody titers were infrequent, consistent with prior experience in unscreened adults. The data suggest that inadvertent vaccination with LAIV in relatively asymptomatic HIV-infected adults would not be associated with frequent significant adverse events. PMID- 10669364 TI - Human immunodeficiency virus-infected persons with mutations conferring resistance to zidovudine show reduced virologic responses to hydroxyurea and stavudine-lamivudine. AB - The baseline predictors of poor virologic response (<0.5 log decrease in plasma virus load) were examined in two 1996 pilot trials of combination nucleoside analogue therapy. One trial examined the addition of hydroxyurea to didanosine therapy; the other examined stavudine-lamivudine in combination. In both, predictors of virologic response included the presence of mutations associated with zidovudine resistance. For hydroxyurea, the odds ratio (OR) of failure to achieve a short-term (4 weeks) virologic response in a bivariate logistic regression model was 30.8 (95% confidence interval [CI], 1.75-543; P=.02) for use of lower dose hydroxyurea (500 mg/day) and 14.7 (95% CI, 1.1-200; P=.04) for the presence of a zidovudine-related mutation. For the stavudine-lamivudine study, the OR of failure to achieve a virologic response at 4 weeks in a multivariate logistic regression model was 23 (95% CI, 2.7-199; P=.004) for the presence of a mutation at codon 215. PMID- 10669365 TI - Effect of interleukin (IL)-15 priming on IL-12 and interferon-gamma production by pathogen-stimulated peripheral blood mononuclear cells from human immunodeficiency virus-seropositive and -seronegative donors. AB - Hypoproduction of the cytokines interleukin (IL)-12 and interferon (IFN)-gamma is thought to contribute to the impaired immunity seen in human immunodeficiency virus (HIV)-infected persons. The effects of priming with IL-15 on the production of IL-12 and IFN-gamma by stimulated peripheral blood mononuclear cells (PBMC) from HIV-seronegative and -seropositive donors were studied. Stimuli included 3 pathogens that commonly infect HIV-positive persons-Cryptococcus neoformans, Candida albicans, and Mycobacterium tuberculosis-plus Staphylococcus aureus. Following IL-15 priming of HIV-negative PBMC, pathogen-stimulated IL-12 and IFN gamma production increased 5-58-fold. However, for the HIV-positive PBMC, IL-15 priming did not lead to significant increases in pathogen-stimulated IL-12 production and caused only modest increases in IFN-gamma production. These data suggest that IL-15 alone may be insufficient to correct the defect in IL-12 and IFN-gamma production in HIV-positive persons. PMID- 10669366 TI - Secretor polymorphism and human immunodeficiency virus infection in Senegalese women. AB - The FUT2 gene encodes the enzyme alpha (1,2) fucosyltransferase, which determines expression of blood-group antigens on mucosal epithelial cell surfaces and in secretions. Homozygotes for a specific stop mutation in FUT2 (nonsecretors) cannot produce this enzyme and thus are unable to express blood group antigens. Nonsecretor status is associated with a decreased risk of several respiratory viral infections. By use of molecular genotyping, 2 populations of Senegalese women were examined for polymorphisms of the FUT2 gene. Among Senegalese commercial sex workers, absence of FUT2 (nonsecretor genotype) was associated with reduced risk of human immunodeficiency virus (HIV) type 1 infection (odds ratio [OR] adjusted for cervical and vaginal infection, 0.18; 95% confidence interval [CI], 0.04-0.90) and HIV-2 infection (adjusted OR, 0.43; 95% CI, 0.13 1.39), although the latter was not statistically significant. Modification of cell surface carbohydrates at mucosal surfaces determined by the FUT2 gene may underlie the protective association against heterosexual HIV infection. PMID- 10669367 TI - Antiretroviral resistance mutations in human immunodeficiency virus type 1 reverse transcriptase and protease from paired cerebrospinal fluid and plasma samples. AB - Twenty-four adults infected with human immunodeficiency virus type 1 (HIV-1) with central nervous system symptoms were studied for antiretroviral resistance mutations in HIV-1 RNA obtained from paired cerebrospinal fluid (CSF) and plasma samples. Paired sequences were obtained from 21 and 13 patients for reverse transcriptase (RT) and for protease, respectively. Mutations conferring resistance to the RT inhibitors zidovudine, lamivudine, or nevirapine were detected in 14 patients, including 11 pretreated and 3 drug-naive subjects. The mutation patterns in the 2 compartments were different in most patients. Genotypic resistance to protease inhibitors was detected in both plasma and CSF from 1 patient treated with multiple protease inhibitors. However, accessory protease inhibitor resistance mutations at polymorphic sites were different in plasma and CSF in several patients. Partially independent evolution of viral quasispecies occurs in plasma and CSF, raising the possibility that compartmentalization of drug resistance may affect response to antiretroviral treatment. PMID- 10669368 TI - Effect of human immunodeficiency virus (HIV) type 1 viral genotype on mother-to child transmission of HIV-1. AB - The objective of this study was to determine whether the maternal infecting human immunodeficiency virus (HIV) type 1 clade affects mother-to-child transmission frequency. Mothers in the mother-to-child HIV-1 transmission study in Nairobi, Kenya, were grouped by HIV-1 status of their first enrolled child: uninfected, perinatally infected, or postnatally infected. Restriction fragment length polymorphism (RFLP) analysis was used to determine HIV-1 viral clades of nested polymerase chain reaction products from HIV-1 protease or p24 genes. When inconclusive, sequencing determined the clade. Clade distributions within the groups were compared. The 3 groups displayed a uniform clade distribution. The predominant clades were A (59%) and D (20%). Clades B, C, F, mixed, and recombinant infections comprised the remainder (21%). No significant association was seen between clades A and D and either frequency or mode of vertical transmission. RFLP analysis revealed 2 clade B infections, 9 mixed, and 5 p24/protease recombinant infections in the study population. PMID- 10669369 TI - Stability of Borrelia burgdorferi outer surface protein C under immune selection pressure. AB - Outer surface protein (Osp) C immune pressure during persistent infection with Borrelia burgdorferi was examined in relation to genetic variation of ospC. Mice were infected with clonal B. burgdorferi sensu stricto (s.s.) N40 or B. afzelii PKo and then were hyperimmunized with homologous recombinant OspC or with decorin binding protein A (DbpA) (controls). After 6 months, B. burgdorferi isolates were subjected to restriction enzyme analysis of the amplified ospC genes and were found to have no differences among 9 B. burgdorferi s.s. N40 and 9 B. afzelii PKo isolates from OspC hyperimmune mice or among 10 B. burgdorferi s.s. N40 and 10 B. afzelii PKo isolates from DbpA hyperimmune mice, compared with input inocula. Comparison of gene sequences among 4 B. burgdorferi s.s. N40 and 9 B. afzelii PKo isolates from OspC-immunized mice revealed no ospC variation from input inocula. Variation in ospC among B. burgdorferi isolates and species during chronic infection is not likely to be an important mechanism for immune evasion. PMID- 10669370 TI - Recombinant human interleukin-11 has anti-inflammatory actions yet does not exacerbate systemic Listeria infection. AB - To determine whether recombinant human (rh) interleukin (IL)-11 disrupts the clearance of microbial pathogens, mice were challenged with Listeria monocytogenes after receiving high-dose rhIL-11, anti-tumor necrosis factor (TNF) monoclonal antibody (MAb), anti-IL-11 MAb, or saline control. The LD50 was not affected by rhIL-11 but was 10-fold lower in the anti-TNF MAb group (P<.001). Plasma IL-6, IL-1beta, and TNF-alpha levels were not different between rhIL-11 treated animals and the control group; however, interferon-gamma levels were significantly reduced by IL-11 treatment (2477 vs. 0 pg/mL, P<.01). Compared with the control group, the quantitative level of L. monocytogenes in hepatic and splenic tissue was unchanged by rhIL-11 but was significantly increased by TNF or IL-11 inhibition. The results indicate that IL-11 down-regulates cytokine production but does not exacerbate systemic infection in the murine Listeria infection model. PMID- 10669371 TI - Pneumococcal vaccine response in cirrhosis and liver transplantation. AB - Cirrhosis is a major risk factor for severe pneumococcal infection, and patients evaluated for liver transplantation routinely receive pneumococcal vaccine. This study followed serologic antibody levels of 45 adults evaluated for transplantation and 13 age-matched control subjects. All received 23-valent pneumococcal polysaccharide vaccine (PPS). Serum anti-PPS levels and antibodies specific for capsular types 3 and 23 were measured by ELISA before and 1 and 6 months after vaccination. Antibody levels for the 25 patients who received transplants also were measured immediately before and 3 months after transplantation. Control subjects had higher IgG responses to the whole vaccine, whereas patients appeared to produce more IgM and IgA. IgA, and possibly IgM levels, also declined faster in patients than in control subjects. All anti-PPS levels were at or below prevaccination baselines by 3 months after transplantation. These data suggest that vaccination with PPS may not be effective for patients during and after liver transplantation. PMID- 10669372 TI - Meningococcal C polysaccharide vaccine induces immunologic hyporesponsiveness in adults that is overcome by meningococcal C conjugate vaccine. AB - Widespread use of meningococcal AC polysaccharide (MACP) vaccines has raised concerns about induction of hyporesponsiveness to C polysaccharide. Whether meningococcal C conjugate (MCC) vaccine overcomes any immunologic refractoriness following MACP vaccination in adults was investigated. University students vaccinated 6 months previously with MACP vaccine were randomized to receive MACP or MCC vaccine, and antibody responses were compared with those of previously unvaccinated students receiving MACP or MCC vaccine. In students primed with MACP vaccine, MCC vaccine induced significantly higher IgG and serum bactericidal antibody levels than did a second dose of MACP vaccine. Responses to a second dose of MACP vaccine were significantly lower than to the first dose. Previous receipt of MACP vaccine reduced serum bactericidal antibody but not IgG responses to MCC vaccine compared with those in previously unvaccinated students. This confirms that MACP vaccine induces immunologic hyporesponsiveness to C polysaccharide in adults, but this can be overcome with MCC vaccine. Repeated vaccination with MACP vaccine may be ineffective, and MCC vaccines should provide better long-term protection. PMID- 10669373 TI - Penetration of clinical isolates of Pseudomonas aeruginosa through MDCK epithelial cell monolayers. AB - Pseudomonas aeruginosa causes both invasive (bacteremic) and chronic noninvasive infections. A simple in vitro system to screen P. aeruginosa clinical isolates for their capacity to penetrate MDCK cell monolayers has been developed. By means of this system, P. aeruginosa clinical isolates, including 32 blood and 45 respiratory isolates, were examined. When monolayers were infected with 3.5x107 cfu of bacteria, significantly more blood (93.7%) than respiratory (54.4%) isolates (P<.001) were detected in the basolateral medium after 3 h. Penetration ability was usually independent of cytotoxicity. Only 8 (4 blood and 4 respiratory) isolates were cytotoxic, possessed exoU, and passed through the monolayer after epithelial cell death, associated with a marked drop in transepithelial electrical resistance. Conversely, noncytotoxic isolates with high penetration ability but without severe epithelial damage were invasive. This system is well suited for screening clinical isolates and their mutants for specific genes conferring the invasiveness phenotype. PMID- 10669374 TI - Poly(sodium 4-styrene sulfonate): an effective candidate topical antimicrobial for the prevention of sexually transmitted diseases. AB - Presently marketed vaginal barrier agents are cytotoxic and damage the vaginal epithelium and natural vaginal flora with frequent use. Novel noncytotoxic agents are needed to protect women from sexually transmitted diseases. One candidate compound is a high-molecular-mass form of soluble poly(sodium 4-styrene sulfonate) (T-PSS). The antimicrobial activity of T-PSS was evaluated in primary culture systems and in a genital herpes murine model. Results obtained indicate that T-PSS is highly effective against herpes simplex viruses, Neisseria gonorrhoeae, and Chlamydia trachomatis in vitro. A 5% T-PSS gel protected 15 of 16 mice from vaginal herpes, compared with 2 of 16 mice treated with a placebo gel. Moreover, T-PSS exhibited little or no cytotoxicity and has an excellent selectivity index. T-PSS is an excellent candidate topical antimicrobial that blocks adherence of herpes simplex virus at low concentrations, inactivates virus at higher concentrations, and exhibits a broad spectrum of antimicrobial activity. PMID- 10669375 TI - Vaccination with FimH adhesin protects cynomolgus monkeys from colonization and infection by uropathogenic Escherichia coli. AB - Escherichia coli FimH adhesin mediates binding to the bladder mucosa. In mice, a FimH vaccine protects against bacterial challenge. In this study, 4 monkeys were inoculated with 100 microgram of FimCH adhesin-chaperone complex mixed with MF59 adjuvant, and 4 monkeys were given adjuvant only intramuscularly. After 2 doses (day 0 and week 4), a booster at 48 weeks elicited a strong IgG antibody response to FimH in the vaccinated monkeys. All 8 monkeys were challenged with 1 mL of 108 E. coli cystitis isolate NU14. Three of the 4 vaccinated monkeys were protected from bacteruria and pyuria; all control monkeys were infected. These findings suggest that a vaccine based on the FimH adhesin of E. coli type 1 pili may have utility in preventing cystitis in humans. PMID- 10669376 TI - Characterization of enterotoxigenic Escherichia coli strains in patients with travelers' diarrhea acquired in Guadalajara, Mexico, 1992-1997. AB - The relationship between enterotoxigenic Escherichia coli (ETEC) and travelers' diarrhea was examined in a high-risk area in 1992-1997. Toxin patterns, colonization-factor antigens (CFAs), and in vitro antimicrobial susceptibility were determined. In total, 928 US students with diarrhea acquired in Guadalajara, Mexico, were screened for enteric pathogens. Diagnosis of ETEC infection was done with oligonucleotide probes. ETEC was isolated in 19.9% of the travelers with diarrhea. CFAs were identified in 51% of the ETEC strains. The highest CFA frequency was observed among heat-stable isolates. Ampicillin, furazolidone, and sulfisoxazole resistance of ETEC increased during the study period. ETEC isolation rates and CFA patterns varied little during the 6 years of the study, which has implications for immunoprophylactic strategies. The finding that differences in the results of ribotyping and plasmid analysis change over time suggests that multiple strains of ETEC were responsible for the illness in the region studied. PMID- 10669377 TI - Inflammatory cytokine mRNA expression during early and persistent Helicobacter pylori infection in nonhuman primates. AB - The role of mononuclear phagocytes in orchestrating the host responses to Helicobacter pylori is inadequately understood. Therefore, gene expression for the monocyte/macrophage-derived cytokines interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha was determined before and during H. pylori infection of rhesus monkeys by use of a highly sensitive quantitative reverse transcriptase polymerase chain reaction. The numbers of molecules of IL-1beta, IL-6, and TNF alpha mRNA in gastric tissue during early infection (7 weeks) significantly exceeded the preinfection numbers (P<.03). Moreover, the numbers of IL-1beta, IL 6, and TNF-alpha mRNA molecules in persistently infected animals (6 years) also were elevated compared with preinfection numbers (P<.02, P=.03, P=.16, respectively). Cytokine gene expression coincided with progressive H. pylori gastritis, confirmed by increased gastritis scores over preinfection scores (P<.005). These findings provide quantitative evidence that H. pylori induces local gene expression of monocyte/macrophage-derived inflammatory cytokines and evokes an innate response in gastric tissue of nonhuman primates. PMID- 10669378 TI - Chlamydia pneumoniae-induced transactivation of the major immediate early promoter of cytomegalovirus: potential synergy of infectious agents in the pathogenesis of atherosclerosis. AB - Both cytomegalovirus (CMV) and Chlamydia pneumoniae have been associated with atherosclerosis. CMV is believed to exist in host tissues in a latent state with periodic reactivation. This study was designed to determine whether C. pneumoniae infection stimulates the expression of CMV genes. Transactivation of the CMV major immediate early promoter (MIEP) is essential for viral gene expression and viral replication. HeLa cells were transfected with a construct containing a reporter gene (chloramphenicol acetyl transferase) controlled by the MIEP. The cells were then infected with Chlamydia at 102-106 infection-forming units (IFU) per well at various times before assay of MIEP activity (72 h after transfection). Peak transactivation occurred 6 h after infection at 104 IFU. C. pneumoniae increased MIEP activity in a dose-response manner; maximal increase was >2-fold. These results suggest that if CMV and C. pneumoniae do indeed contribute to atherosclerosis, their copresence may synergistically contribute to it. PMID- 10669379 TI - Stimulation of macrophage inflammatory protein-1alpha, macrophage inflammatory protein-1beta, and RANTES by Candida albicans and Cryptococcus neoformans in peripheral blood mononuclear cells from persons with and without human immunodeficiency virus infection. AB - The beta-chemokine macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES are critical for recruitment of inflammatory cells into infected tissue. Moreover, by binding to the human immunodeficiency virus (HIV) coreceptor CCR5, release of these chemokines could influence the course of HIV infection. beta chemokine gene expression and release was determined by ELISA and RNase protection assay, respectively, in peripheral blood mononuclear cells (PBMC) from HIV-negative and -positive persons stimulated with Candida albicans and Cryptococcus neoformans, 2 fungi common in HIV-infected persons. Gene expression and/or release of all 3 chemokines was seen in response to both fungi although C. albicans was more potent than C. neoformans. Fungal stimulated chemokine production by HIV-positive PBMC was similar to that in HIV-negative PBMC, suggesting that the scant inflammatory response often seen in AIDS patients with cryptococcosis and candidiasis is not secondary to suboptimal beta-chemokine release. PMID- 10669380 TI - Visceral leishmanicidal activity of hexadecylphosphocholine (miltefosine) in mice deficient in T cells and activated macrophage microbicidal mechanisms. AB - Hexadecylphosphocholine (miltefosine), a membrane-active alkylphospholipid, may be the first effective oral agent for visceral leishmaniasis, an intracellular protozoal infection of tissue macrophages. In vitro, miltefosine stimulates T cells and macrophages to respond to and secrete activating cytokines, including interferon (IFN)-gamma, and enhances macrophage production of microbicidal reactive nitrogen and oxygen intermediates (RNIs and ROIs, respectively). To determine whether these effects mediate miltefosine's in vivo leishmanicidal efficacy, genetically deficient mice were infected with Leishmania donovani. Intracellular visceral killing was retained in mice lacking or deficient in T cells, endogenous IFN-gamma, and macrophage generation of leishmanicidal RNIs and ROIs. Although mutant mice responded to miltefosine in the absence of tissue granulomas, treatment enhanced granuloma assembly in normal animals. These results suggest that miltefosine's visceral leishmanicidal effect does not require host T cell-dependent or activated macrophage-mediated mechanisms; thus, this agent may potentially be useful in treating T cell-deficient patients with kala-azar. PMID- 10669382 TI - Reply. PMID- 10669381 TI - Mastitis and human immunodeficiency virus transmission: chemokines and maternal monocytes. PMID- 10669383 TI - Preliminary falsification of EIA screening is cost-effective in the two-step serodiagnosis of lyme disease. PMID- 10669385 TI - Reply. PMID- 10669384 TI - TT virus: evidence for transplacental transmission. PMID- 10669386 TI - Neglected opportunities. PMID- 10669388 TI - Reply. PMID- 10669387 TI - Helicobacter pylori, lifestyle risk factors, and adenocarcinoma of the esophagus. PMID- 10669389 TI - Reemergence of invasive Haemophilus influenzae type b disease in Alaska: Is it because of vaccination with polyribosylribitol phosphate outer membrane protein complex (PRP-OMPC) or failure to vaccinate with PRP-OMPC? PMID- 10669390 TI - Reply. PMID- 10669391 TI - Mortality in serologically unconfirmed Mediterranean spotted fever. PMID- 10669392 TI - Reply. PMID- 10669393 TI - Oropharyngeal Candida colonization and human immunodeficiency virus type 1 infection. PMID- 10669394 TI - Reply. PMID- 10669396 TI - Bioconversion of D-galactose into D-tagatose by expression of L-arabinose isomerase. AB - D-Tagatose is a potential bulking agent in food as a non-calorific sweetener. To produce D-tagatose from cheaper resources, plasmids harbouring the L-arabinose isomerase gene (araA) from Escherichia coli, Bacillus subtilis and Salmonella typhimurium were constructed because L-arabinose isomerase was suggested previously as an enzyme that mediates the bioconversion of galactose into tagatose as well as that of arabinose to ribulose. The constructed plasmids were named pTC101, pTC105 and pTC106, containing araA from E. coli, B. subtilis and S. typhimurium respectively. In the cultures of recombinant E. coli with pTC101, pTC105 and pTC106, tagatose was produced from galactose in 9.9, 7.1 and 6.9% yields respectively. The enzyme extract of E. coli with the plasmid pTC101 also converted galactose into tagatose with a 96.4% yield. PMID- 10669397 TI - Characterization of N-linked oligosaccharides bearing sialyl lewis x moieties on an alternatively glycosylated form of soluble complement receptor type 1 (sCR1). AB - We sought to produce a complement inhibitory protein possessing oligosaccharides specifically modified to contain the sialyl Lewis x (sLe(x)) moiety. This modified glycoprotein could combine anti-complement activity with the ability to inhibit selectin-mediated interactions and concentrate this activity to sites of activated endothelium where selectins are upregulated. Soluble complement receptor type 1 (sCR1), previously shown to be effective in inhibiting the complement cascade, was produced in a cell line capable of adding fucose to N linked oligosaccharides in the alpha1-3 linkage, which is necessary for sLe(x) glycosylation. The glycoprotein purified from these cells was designated sCR1sLe(x), and may prove to be more effective than sCR1 in some clinical applications. Detailed analysis and characterization of sCR1sLe(x) was performed to confirm that the N-linked oligosaccharides possessed sLe(x) moieties and also to determine the extent of sLe(x) glycosylation. The glycoproteins were characterized by oligosaccharide profiling, sequencing, linkage analysis and quantified by differential enzymic digestion, using fluorophore-assisted carbohydrate electrophoresis. The major glycans were identified as biantennary oligosaccharides (including sialylated and non-core fucosylated glycans). The linkages of sialic acid and the branched fucose were analysed by digestion with linkage-specific enzymes and subsequent separation by electrophoresis. All data were consistent with the presence of sLe(x) moieties on the N-linked oligosaccharides of sCR1sLe(x). sCR1sLe(x) is a prime example of a recombinant protein expressed with oligosaccharides engineered for a specific biological function, and produced using a commercially viable method. PMID- 10669398 TI - Affinity adsorbents for the vancomycin group of antibiotics. AB - The vancomycin group of antibiotics kill Gram-positive bacteria by binding to nascent bacterial cell-wall peptidoglycan bearing the C-terminal sequence-D-Ala-D Ala. In this paper, affinity adsorbents for the vancomycin group of antibiotics were prepared by immobilizing the peptidoglycan analogues -D-Ala-D-Ala, -succinyl D-Ala and -succinyl-Gly on to crosslinked poly(N, N-dimethylacrylamide). The adsorption capacities of the three adsorbents for demethylvancomycin were 0.59, 0.35 and 0.29 mmol/g, respectively. The adsorption capacity of the adsorbent with D-Ala-D-Ala for vancomycin was 0.53 mmol/g. In contrast, the adsorbent bearing succinyl-L-Ala hardly adsorbed demethylvancomycin. Aqueous sodium carbonate (0.4 M)/acetonitrile (7/3, v/v) completely desorbed demethylvancomycin adsorbed on the adsorbents. PMID- 10669399 TI - Selecting and expressing protective single-chain Fv fragment to stabilize L asparaginase against inactivation by trypsin. AB - Four non-inhibitory specific single-chain Fv (sc Fv) fragments directed against L asparaginase (ASNase) of Escherichia coli were selected from a synthetic phage display scFv library. The scFv46 fragment could enhance the resistance of ASNase to trypsin proteolysis, with 70% of the initial ASNase activity present after the ASNase-scFv46 complex had been treated with trypsin for 30 min at 37 degrees C, whereas little residual activity was detected without the scFv46 fragment. The scFv46 gene was cloned to an expression vector pET-21a and expressed at high levels (about 45% of total cell protein) in E. coli BL21 (DE3) as inclusion bodies. The refolded and purified scFv46 fragment was proved to protect ASNase, and the protective effect was further confirmed by SDS/PAGE. It was found that under optimum conditions of molar ratio of scFv to ASNase, incubation time and temperature, the residual activity of the ASNase-scFv46 complex could reach about 78% after treatment with trypsin for 30 min at 37 degrees C. The results demonstrated that scFv fragments prepared by phage-antibody library technology could be used to protect target proteins. PMID- 10669400 TI - Conformational issues in the characterization of proteins. AB - The conformation of a protein refers to the three-dimensional arrangement of its constituent atoms. Since expression of the biological activity of a protein depends on its conformation, it is clear that full characterization of a protein involves an understanding of its three-dimensional structure. This review outlines the principal techniques for determining the conformation of a protein, describes the crucial role played by the flexibility of proteins, and gives an account of current theories of the mechanisms by which proteins fold. A final section deals with strategies that can be adopted to preserve the conformational integrity of proteins; an aspect that is of increasing importance as a greater number of proteins are finding applications in industrial processes and as therapeutic and diagnostic agents. PMID- 10669401 TI - Relationship between conformational stability and lyophilization-induced structural changes in chymotrypsin. AB - The relationship between protein conformational stability in aqueous solution and the magnitude of lyophilization-induced structural changes was investigated employing alpha- and gamma-chymotrypsin. As a measure of the conformational stability the melting temperature T(m) was determined in distilled water at various pH values. The proteins were then lyophilized from those pH values where the conformational stability was maximum (pH 4.5) and minimum (pH 7.8). Protein secondary structure was quantitatively determined utilizing Fourier-transform infrared spectroscopy employing two regions sensitive to protein structure, the amide-I (1600-1700 cm(-1)) and amide-III (1215-1335 cm(-1)). Lyophilization induced significant structural alterations in both proteins, characterized by a slight decrease in the alpha-helix and a significant increase in the beta-sheet content. However, regardless of the pH from which the proteins were lyophilized, the secondary structures in the solid state were indistinguishable. This result shows that there is no relationship between the conformational stability in aqueous solution and the magnitude of lyophilization-induced structural changes. We also investigated whether lyoprotectants could minimize lyophilization-induced structural changes by increasing protein conformational stability in aqueous solution. After having identified trehalose as being efficient in largely preventing lyophilization-induced structural alterations, we conducted co lyophilization experiments from various pH values. The results obtained exclude any contribution from increased protein conformational stability caused by the additive in aqueous solution to the beneficial structural preservation upon lyophilization. This can be understood because the dehydration and not the freezing process, as shown in an air-drying experiment, mainly causes protein structural alterations. PMID- 10669402 TI - Efficient utilization of starch by a recombinant strain of Saccharomyces cerevisiae producing glucoamylase and isoamylase. AB - Two plasmids, designated pRTI and pTI, were constructed to allow the integration of a bacterial isoamylase gene (iso) into Saccharomyces cerevisiae G23-8 chromosome. The integrative plasmid pRTI comprises the iso gene from Pseudomonas amyloderamosa, a portion of S. cerevisiae ribosomal DNA (rDNA), S. cerevisiae trp1 gene deficient in promoter and the bacterial vector pSP72. The structure of plasmid pTI is similar to that of pRTI, except that it lacks an rDNA segment. The Aspergillus awamori glucoamylase and P. amyloderamosa isoamylase genes were expressed in the recombinant strain of S. cerevisiae under the control of the yeast alcohol dehydrogenase gene (adh1) promoter. Southern-blot analysis showed that these plasmids were integrated into the yeast chromosome in tandem repeat and dispersion copies. The recombinant strains could assimilate starch more efficiently than the recipient strain with a conversion rate of greater than 95%. PMID- 10669403 TI - Endoxylanase II from Trichoderma reesei has several isoforms with different isoelectric points. AB - Two minor xylanases present in Trichoderma reesei Rut C30 cultivation broth were purified as a mixture using ion-exchange, hydrophobic-interaction and gel chromatography. The purified enzyme preparation contained two active xylanases with pI values of 7.1 and 8.1. Both components had a molecular mass of 20 kDa. The purified xylanase preparation exhibited properties very similar to those of the previously isolated XYL II (pI 9.0) of T. reesei Rut C30. The activity and stability properties, apparent kinetic parameters as well as the titration curve forms were similar. The major difference in enzymic properties was the significantly lower specific activity of the pI-7.1+8.1 xylanase mixture (3350 nkat/mg) compared with the specific activity of XYL II (13500 nkat/mg). Amino acid sequences of tryptic peptides (34% of the total amino acid sequence was determined) were identical to the amino acid sequence of XYL II. Furthermore, in vitro modification of the pI-9.0 form of XYL II to pI-8.1 and pI-7.1 forms was demonstrated. Thus the purified xylanase preparation most probably contained two modified forms of XYL II. The primary amino acid sequence of XYL II contains 28 glutamine and asparagine residues and theoretically deamination of one of them lowers the pI to 8.06 and deamination of two amino acids lowers the pI to 7.02. PMID- 10669404 TI - Conformational issues in the characterization of proteins. PMID- 10669405 TI - Phase-specific optimization of multiple endotoxin-protein production with genetically engineered Bacillus thuringiensis. AB - An optimization approach was designed to specifically study the toxin-expression phase of the fermentation process of a genetically engineered Bacillus thuringiensis strain expressing dual toxin proteins (CryI and CryIII). The study has resulted in the discovery of important nutrient and process factors affecting toxin-protein yield. The results show that the existence of nitrogen sources in the medium during the toxin-expression phase is detrimental to the toxin-protein expression, while a high carbon-source level (40 g/l) encouraged protein expression. The study also suggests that the depletion of nitrogen source is the trigger for B. thuringiensis to initiate sporulation and toxin expression. A temperature setting of 28 degrees C for B. thuringiensis fermentation processes is optimal for protein yield, and reduces the oxygen requirement. It was found that the optimal conditions for spore yield and for toxin-protein yield were not the same, even though sporulation and toxin formation proceed simultaneously during the fermentation process. Scale-up studies were also conducted to confirm the optimal conditions obtained from a small-scale optimization study. PMID- 10669406 TI - Quantization of multiparticle auger rates in semiconductor quantum dots AB - We have resolved single-exponential relaxation dynamics of the 2-, 3-, and 4 electron-hole pair states in nearly monodisperse cadmium selenide quantum dots with radii ranging from 1 to 4 nanometers. Comparison of the discrete relaxation constants measured for different multiple-pair states indicates that the carrier decay rate is cubic in carrier concentration, which is characteristic of an Auger process. We observe that in the quantum-confined regime, the Auger constant is strongly size-dependent and decreases with decreasing the quantum dot size as the radius cubed. PMID- 10669407 TI - Observation of antiferromagnetic domains in epitaxial thin films AB - Antiferromagnetic domains in an epitaxial thin film, LaFeO(3) on SrTiO(3)(100), were observed using a high-spatial-resolution photoelectron emission microscope with contrast generated by the large x-ray magnetic linear dichroism effect at the multiplet-split L edge of Fe. The antiferromagnetic domains are linked to 90 degrees twinned crystallographic regions in the film. The Neel temperature of the thin film is reduced by 70 kelvin relative to the bulk material, and this reduction is attributed to epitaxial strain. These studies open the door for a microscopic understanding of the magnetic coupling across antiferromagnetic ferromagnetic interfaces. PMID- 10669408 TI - Molecules in a bose-einstein condensate AB - State-selected rubidium-87 molecules were created at rest in a dilute Bose Einstein condensate of rubidium-87 atoms with coherent free-bound stimulated Raman transitions. The transition rate exhibited a resonance line shape with an extremely narrow width as small as 1.5 kilohertz. The precise shape and position of the resonance are sensitive to the mean-field interactions between the molecules and the atomic condensate. As a result, we were able to measure the molecule-condensate interactions. This method allows molecular binding energies to be determined with unprecedented accuracy and is of interest as a mechanism for the generation of a molecular Bose-Einstein condensate. PMID- 10669409 TI - Zener model description of ferromagnetism in zinc-blende magnetic semiconductors AB - Ferromagnetism in manganese compound semiconductors not only opens prospects for tailoring magnetic and spin-related phenomena in semiconductors with a precision specific to III-V compounds but also addresses a question about the origin of the magnetic interactions that lead to a Curie temperature (T(C)) as high as 110 K for a manganese concentration of just 5%. Zener's model of ferromagnetism, originally proposed for transition metals in 1950, can explain T(C) of Ga(1 )(x)Mn(x)As and that of its II-VI counterpart Zn(1-)(x)Mn(x)Te and is used to predict materials with T(C) exceeding room temperature, an important step toward semiconductor electronics that use both charge and spin. PMID- 10669410 TI - Ambipolar pentacene field-effect transistors and inverters. AB - Organic field-effect transistors based on pentacene single crystals, prepared with an amorphous aluminum oxide gate insulator, are capable of ambipolar operation and can be used for the preparation of complementary inverter circuits. The field-effect mobilities of carriers in these transistors increase from 2.7 and 1.7 square centimeters per volt per second at room temperature up to 1200 and 320 square centimeters per volt per second at low temperatures for hole and electron transport, respectively, following a power-law dependence. The possible simplification of the fabrication process of complementary logic circuits with these transistors, together with the high carrier mobilities, may be seen as another step toward applications of plastic electronics. PMID- 10669411 TI - CsBi(4)Te(6): A high-performance thermoelectric material for low-temperature applications AB - Thermoelectric (Peltier) heat pumps are capable of refrigerating solid or fluid objects, and unlike conventional vapor compressor systems, they can be miniaturized without loss of efficiency. More efficient thermoelectric materials need to be identified, especially for low-temperature applications in electronics and devices. The material CsBi(4)Te(6) has been synthesized and its properties have been studied. When doped appropriately, it exhibits a high thermoelectric figure of merit below room temperature (ZT(max) approximately 0.8 at 225 kelvin). At cryogenic temperatures, the thermoelectric properties of CsBi(4)Te(6) appear to match or exceed those of Bi(2-x)Sb(x)Te(3-y)Se(y) alloys. PMID- 10669412 TI - Physics of iron at Earth's core conditions AB - The bulk properties of iron at the pressure and temperature conditions of Earth's core were determined by a method that combines first-principles and classical molecular dynamic simulations. The theory indicates that (i) the iron melting temperature at inner-core boundary (ICB) pressure (330 gigapascals) is 5400 (+/ 400) kelvin; (ii) liquid iron at ICB conditions is about 6% denser than Earth's outer core; and (iii) the shear modulus of solid iron close to its melting line is 140 gigapascals, consistent with the seismic value for the inner core. These results reconcile melting temperature estimates based on sound velocity shock wave data with those based on diamond anvil cell experiments. PMID- 10669413 TI - Surface expression of HLA-E, an inhibitor of natural killer cells, enhanced by human cytomegalovirus gpUL40. AB - The nonclassical major histocompatibility complex (MHC) class I molecule HLA-E inhibits natural killer (NK) cell-mediated lysis by interacting with CD94/NKG2A receptors. Surface expression of HLA-E depends on binding of conserved peptides derived from MHC class I molecules. The same peptide is present in the leader sequence of the human cytomegalovirus (HCMV) glycoprotein UL40 (gpUL40). It is shown that, independently of the transporter associated with antigen processing, gpUL40 can up-regulate expression of HLA-E, which protects targets from NK cell lysis. While classical MHC class I molecules are down-regulated, HLA-E is up regulated by HCMV. Induction of HLA-E surface expression by gpUL40 may represent an escape route for HCMV. PMID- 10669414 TI - Localization of the G protein betagamma complex in living cells during chemotaxis. AB - Gradients of chemoattractants elicit signaling events at the leading edge of a cell even though chemoattractant receptors are uniformly distributed on the cell surface. In highly polarized Dictyostelium discoideum amoebas, membrane associated betagamma subunits of heterotrimeric guanine nucleotide-binding proteins (G proteins) were localized in a shallow anterior-posterior gradient. A uniformly applied chemoattractant generated binding sites for pleckstrin homology (PH) domains on the inner surface of the membrane in a pattern similar to that of the Gbetagamma subunits. Loss of cell polarity resulted in uniform distribution of both the Gbetagamma subunits and the sensitivity of PH domain recruitment. These observations indicate that Gbetagamma subunits are not sufficiently localized to restrict signaling events to the leading edge but that their distribution may determine the relative chemotactic sensitivity of polarized cells. PMID- 10669415 TI - Polarization of chemoattractant receptor signaling during neutrophil chemotaxis. AB - Morphologic polarity is necessary for chemotaxis of mammalian cells. As a probe of intracellular signals responsible for this asymmetry, the pleckstrin homology domain of the AKT protein kinase (or protein kinase B), tagged with the green fluorescent protein (PHAKT-GFP), was expressed in neutrophils. Upon exposure of cells to chemoattractant, PHAKT-GFP is recruited selectively to membrane at the cell's leading edge, indicating an internal signaling gradient that is much steeper than that of the chemoattractant. Translocation of PHAKT-GFP is inhibited by toxin-B from Clostridium difficile, indicating that it requires activity of one or more Rho guanosine triphosphatases. PMID- 10669416 TI - Function of PI3Kgamma in thymocyte development, T cell activation, and neutrophil migration. AB - Phosphoinositide 3-kinases (PI3Ks) regulate fundamental cellular responses such as proliferation, apoptosis, cell motility, and adhesion. Viable gene-targeted mice lacking the p110 catalytic subunit of PI3Kgamma were generated. We show that PI3Kgamma controls thymocyte survival and activation of mature T cells but has no role in the development or function of B cells. PI3Kgamma-deficient neutrophils exhibited severe defects in migration and respiratory burst in response to heterotrimeric GTP-binding protein (G protein)-coupled receptor (GPCR) agonists and chemotactic agents. PI3Kgamma links GPCR stimulation to the formation of phosphatidylinositol 3,4,5-triphosphate and the activation of protein kinase B, ribosomal protein S6 kinase, and extracellular signal-regulated kinases 1 and 2. Thus, PI3Kgamma regulates thymocyte development, T cell activation, neutrophil migration, and the oxidative burst. PMID- 10669418 TI - Central role for G protein-coupled phosphoinositide 3-kinase gamma in inflammation. AB - Phosphoinositide 3-kinase (PI3K) activity is crucial for leukocyte function, but the roles of the four receptor-activated isoforms are unclear. Mice lacking heterotrimeric guanine nucleotide-binding protein (G protein)-coupled PI3Kgamma were viable and had fully differentiated neutrophils and macrophages. Chemoattractant-stimulated PI3Kgamma-/- neutrophils did not produce phosphatidylinositol 3,4,5-trisphosphate, did not activate protein kinase B, and displayed impaired respiratory burst and motility. Peritoneal PI3Kgamma-null macrophages showed a reduced migration toward a wide range of chemotactic stimuli and a severely defective accumulation in a septic peritonitis model. These results demonstrate that PI3Kgamma is a crucial signaling molecule required for macrophage accumulation in inflammation. PMID- 10669417 TI - Roles of PLC-beta2 and -beta3 and PI3Kgamma in chemoattractant-mediated signal transduction. AB - The roles of phosphoinositide 3-kinase (PI3K) and phospholipase C (PLC) in chemoattractant-elicited responses were studied in mice lacking these key enzymes. PI3Kgamma was required for chemoattractant-induced production of phosphatidylinositol 3,4,5-trisphosphate [PtdIns (3,4,5)P3] and has an important role in chemoattractant-induced superoxide production and chemotaxis in mouse neutrophils and in production of T cell-independent antigen-specific antibodies composed of the immunoglobulin lambda light chain (TI-IglambdaL). The study of the mice lacking PLC-beta2 and -beta3 revealed that the PLC pathways have an important role in chemoattractant-mediated production of superoxide and regulation of protein kinases, but not chemotaxis. The PLC pathways also appear to inhibit the chemotactic activity induced by certain chemoattractants and to suppress TI-IglambdaL production. PMID- 10669419 TI - Requirement for DARPP-32 in progesterone-facilitated sexual receptivity in female rats and mice. AB - DARPP-32, a dopamine- and adenosine 3',5'-monophosphate (cAMP)-regulated phosphoprotein (32 kilodaltons in size), is an obligate intermediate in progesterone (P)-facilitated sexual receptivity in female rats and mice. The facilitative effect of P on sexual receptivity in female rats was blocked by antisense oligonucleotides to DARPP-32. Homozygous mice carrying a null mutation for the DARPP-32 gene exhibited minimal levels of P-facilitated sexual receptivity when compared to their wild-type littermates. P significantly increased hypothalamic cAMP levels and cAMP-dependent protein kinase activity. These increases were not inhibited by a D1 subclass dopamine receptor antagonist. P also enhanced phosphorylation of DARPP-32 on threonine 34 in the hypothalamus of mice. DARPP-32 activation is thus an obligatory step in progestin receptor regulation of sexual receptivity in rats and mice. PMID- 10669420 TI - Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome. AB - The deleterious effects of ethanol on the developing human brain are poorly understood. Here it is reported that ethanol, acting by a dual mechanism [blockade of N-methyl-D-aspartate (NMDA) glutamate receptors and excessive activation of GABA(A) receptors], triggers widespread apoptotic neurodegeneration in the developing rat forebrain. Vulnerability coincides with the period of synaptogenesis, which in humans extends from the sixth month of gestation to several years after birth. During this period, transient ethanol exposure can delete millions of neurons from the developing brain. This can explain the reduced brain mass and neurobehavioral disturbances associated with human fetal alcohol syndrome. PMID- 10669421 TI - Evidence for DNA loss as a determinant of genome size. AB - Eukaryotic genome sizes range over five orders of magnitude. This variation cannot be explained by differences in organismic complexity (the C value paradox). To test the hypothesis that some variation in genome size can be attributed to differences in the patterns of insertion and deletion (indel) mutations among organisms, this study examines the indel spectrum in Laupala crickets, which have a genome size 11 times larger than that of Drosophila. Consistent with the hypothesis, DNA loss is more than 40 times slower in Laupala than in Drosophila. PMID- 10669423 TI - Nicotine addiction. PMID- 10669422 TI - Conservation and novelty in the evolution of cell adhesion and extracellular matrix genes. AB - New proteins and modules have been invented throughout evolution. Gene "birth dates" in Caenorhabditis elegans range from the origins of cellular life through adaptation to a soil habitat. Possibly half are "metazoan" genes, having arisen sometime between the yeast-metazoan and nematode-chordate separations. These include basement membrane and cell adhesion molecules implicated in tissue organization. By contrast, epithelial surfaces facing the environment have specialized components invented within the nematode lineage. Moreover, interstitial matrices were likely elaborated within the vertebrate lineage. A strategy for concerted evolution of new gene families, as well as conservation of adaptive genes, may underlie the differences between heterochromatin and euchromatin. PMID- 10669424 TI - Voluntary organisations: from Cinderella to white knight? PMID- 10669425 TI - From CME to CPD: getting better at getting better? PMID- 10669426 TI - Should doctors get CME points for reading? PMID- 10669427 TI - The changing face of refractive surgery. PMID- 10669428 TI - Doctors told to treat nicotine addiction as a disease. PMID- 10669429 TI - Video guide to suicide is shown on television. PMID- 10669431 TI - In brief PMID- 10669430 TI - Breast cancer researcher accused of serious scientific misconduct. PMID- 10669432 TI - Neurosurgical units working beyond safe capacity. PMID- 10669433 TI - Solicitor in litigation case condemns tobacco industry. PMID- 10669434 TI - Oral sex may be important risk factor for HIV infection. PMID- 10669435 TI - Canada faces healthcare crisis PMID- 10669436 TI - Medical research site in Berlin threatened. PMID- 10669437 TI - Convalescent homes to make a comeback. PMID- 10669438 TI - Milburn sets up inquiry into Shipman case. PMID- 10669439 TI - BMA accused of contempt of court. PMID- 10669440 TI - WHO study examines teenage health in 28 countries. PMID- 10669441 TI - Radiation of the arteries can reduce narrowing PMID- 10669442 TI - UK launches initiative on global health PMID- 10669443 TI - Prospective investigation of transfusion transmitted infection in recipients of over 20 000 units of blood. TTI Study Group. AB - OBJECTIVES: To follow up recipients of 20 000 units of blood to identify any transmissions of infections through blood transfusion. DESIGN: Follow up study of recipients of transfusion. SETTING: 22 hospitals in north London. PARTICIPANT: Adult patients who had recently been transfused. MAIN OUTCOME MEASURES: Patients had further blood samples taken at 9 months that were tested for markers of hepatitis B and C and HIV and human T cell leukaemia/lymphoma virus type I or II (HTLV) infections. Recent infections were distinguished from pre-existing infections by comparison with blood samples taken before transfusion. RESULTS: 9220 patients were recruited, and 5579 recipients of 21 923 units of blood were followed up. No transfusion transmitted infections were identified. The incidence of transfusion transmitted infections was 0 in 21 043 units (95% confidence interval for risk 0 to 1 in 5706 recipients) for hepatitis B; 0 in 21 800 units (0 to 1 in 5911 recipients) for hepatitis C; 0 in 21 923 units (0 to 1 in 5944 recipients) for HIV; and 0 in 21 902 units (0 to 1 in 5939 recipients) for human T cell leukaemia/lymphoma virus. Three patients acquired hepatitis B during or after hospital admission but not through transfusion; 176 (3%) had pre-existing hepatitis B infection. Sixteen (0.29%) patients had hepatitis C, and five (0.09%) had human T cell leukaemia/lymphoma virus. CONCLUSIONS: The current risk of transfusion transmitted infections in the United Kingdom is very small, though hospital acquired infections may arise from sources other than transfusion. A considerable proportion of patients have pre-existing infections. PMID- 10669444 TI - Psychological consequences for parents of false negative results on prenatal screening for Down's syndrome: retrospective interview study. AB - OBJECTIVE: To determine the psychological consequences for parents of children with Down's syndrome of having received a false negative result on prenatal screening. DESIGN: Comparison of adjustment of parents who received a false negative result with that of parents not offered a test and those who declined a test. SETTING: Parents were interviewed in their own homes. PARTICIPANTS: Parents of 179 children with Down's syndrome (mean age 4 (range 2-6) years). MAIN OUTCOME MEASURES: Anxiety, depression, parenting stress, attitudes towards the child, and attributions of blame for the birth of the affected child. RESULTS: Overall, regardless of screening history, parents adjusted well to having a child with Down's syndrome. Compared with mothers who declined a test, mothers in the false negative group had higher parenting stress (mean score 81.2 v 71.8, P=0.016, 95% confidence interval for the difference 1.8 to 17.0) and more negative attitudes towards their children (124.9 v 134.2, P=0. 009, -16.2 to -2.4). Fathers in the false negative group had higher parenting stress test scores (77.8 v 70.0, P=0.046, 1.5 to 14.2) than fathers not offered a test. Mothers in the false negative group were more likely to blame others for the outcome than mothers who had not been offered the test (28% v 13%, P=0.032, 3% to 27%). Mothers and fathers in the false negative group were more likely to blame others for this outcome than parents who had declined a test (mothers 28% v 0%, P=0.001, 19% to 37%; fathers 27% v 0%, P=0.004, 17% to 38%). Blaming others was associated with poorer adjustment for mothers and fathers. CONCLUSIONS: A false negative result on prenatal screening seems to have a small adverse effect on parental adjustment evident two to six years after the birth of an affected child. PMID- 10669445 TI - Exposure to foodborne and orofecal microbes versus airborne viruses in relation to atopy and allergic asthma: epidemiological study. AB - OBJECTIVE: To investigate if markers of exposure to foodborne and orofecal microbes versus airborne viruses are associated with atopy and respiratory allergies. DESIGN: Retrospective case-control study. PARTICIPANTS: 240 atopic cases and 240 non-atopic controls from a population sample of 1659 participants, all Italian male cadets aged 17-24. SETTING: Air force school in Caserta, Italy. MAIN OUTCOME MEASURES: Serology for Toxoplasma gondii, Helicobacter pylori, hepatitis A virus, measles, mumps, rubella, chickenpox, cytomegalovirus, and herpes simplex virus type 1; skin sensitisation and IgE antibodies to relevant airborne allergens; total IgE concentration; and diagnosis of allergic asthma or rhinitis. RESULTS: Compared with controls there was a lower prevalence of T gondii (26% v 18%, P=0.027), hepatitis A virus (30% v 16%, P=0.004), and H pylori (18% v 15%, P=0.325) in atopic participants. Adjusted odds ratios of atopy decreased with a gradient of exposure to H pylori, T gondii, and hepatitis A virus (none, odds ratio 1; one, 0. 70; two or three, 0.37; P for trend=0.000045) but not with cumulative exposure to the other viruses. Conversely, total IgE concentration was not independently associated with any infection. Allergic asthma was rare (1/245, 0.4%) and allergic rhinitis infrequent (16/245, 7%) among the participants (245/1659) exposed to at least two orofecal and foodborne infections (H pylori, T gondii, hepatitis A virus). CONCLUSION: Respiratory allergy is less frequent in people heavily exposed to orofecal and foodborne microbes. Hygiene and a westernised, semisterile diet may facilitate atopy by influencing the overall pattern of commensals and pathogens that stimulate the gut associated lymphoid tissue thus contributing to the epidemic of allergic asthma and rhinitis in developed countries. PMID- 10669446 TI - Reanalysis of epidemiological evidence on lung cancer and passive smoking. AB - OBJECTIVE: To assess the epidemiological evidence for an increase in the risk of lung cancer resulting from exposure to environmental tobacco smoke. DESIGN: Reanalysis of 37 published epidemiological studies previously included in a meta analysis allowing for the possibility of publication bias. MAIN OUTCOME MEASURE: Relative risk of lung cancer among female lifelong non-smokers, according to whether her partner was a current smoker or a lifelong non-smoker. RESULTS: If it is assumed that all studies that have ever been carried out are included, or that those selected for review are truly representative of all such studies, then the estimated excess risk of lung cancer is 24%, as previously reported (95% confidence interval 13% to 36%, P<0.001). However, a significant correlation between study outcome and study size suggests the presence of publication bias. Adjustment for such bias implies that the risk has been overestimated. For example, if only 60% of studies have been included, the estimate of excess risk falls from 24% to 15%. CONCLUSION: A modest degree of publication bias leads to a substantial reduction in the relative risk and to a weaker level of significance, suggesting that the published estimate of the increased risk of lung cancer associated with environmental tobacco smoke needs to be interpreted with caution. PMID- 10669447 TI - A randomised controlled trial and economic evaluation of a referrals facilitator between primary care and the voluntary sector. AB - OBJECTIVES: To compare outcome and resource utilisation among patients referred to the Amalthea Project, a liaison organisation that facilitates contact between voluntary organisations and patients in primary care, with patients receiving routine general practitioner care. DESIGN: Randomised controlled trial with follow up at one and four months. SETTING: 26 general practices in Avon. PARTICIPANTS: 161 patients identified by their general practitioner as having psychosocial problems. MAIN OUTCOME MEASURES: Primary outcomes were psychological wellbeing (assessed with the hospital anxiety and depression scale) and social support (assessed using the Duke-UNC functional social support questionnaire). Secondary outcomes were quality of life measures (the Dartmouth COOP/WONCA functional health assessment charts and the delighted-terrible faces scale), cost of contacts with the primary healthcare team and Amalthea Project, cost of prescribing in primary care, and cost of referrals to other agencies, over four months. RESULTS: The Amalthea group showed significantly greater improvements in anxiety (average difference between groups after adjustment for baseline -1.9, 95% confidence interval -3.0 to -0.7), other emotional feelings (average adjusted difference -0.5, -0.8 to -0.2), ability to carry out everyday activities (-0.5, 0.8 to -0.2), feelings about general health (-0.4, -0.7 to -0.1), and quality of life (-0.5, -0.9 to -0.1). No difference was detected in depression or perceived social support. The mean cost was significantly greater in the Amalthea arm than the general practitioner care arm ( pound153 v pound133, P=0. 025). CONCLUSION: Referral to the Amalthea Project and subsequent contact with the voluntary sector results in clinically important benefits compared with usual general practitioner care in managing psychosocial problems, but at a higher cost. PMID- 10669449 TI - In the line of duty PMID- 10669448 TI - Regular review: tumour markers in malignancies. PMID- 10669450 TI - ABC of heart failure. Management: diuretics, ACE inhibitors, and nitrates. PMID- 10669451 TI - Continuing medical education and continuing professional development: international comparisons. PMID- 10669453 TI - Tuberculosis in prisons in countries with high prevalence. PMID- 10669452 TI - Are generalists still needed in a specialised world? The renaissance of general surgery. PMID- 10669454 TI - Human population growth. Rich countries need education on resource conservation. PMID- 10669455 TI - Registering trials is not enough to counter perceived irrelevance of much research. PMID- 10669456 TI - Coping with winter bed crises. Crises do not just happen in winter. PMID- 10669457 TI - Prospective risk of stillbirth. Study's results are flawed by reliance on cumulative prospective risk. PMID- 10669458 TI - Treatment of venous leg ulcers. Nice study, pity about the sample size. PMID- 10669459 TI - Stages of change model for smoking prevention and cessation in schools. Authors applied adult dose for smoking to adolescents when smoking behaviour is different in the two. PMID- 10669460 TI - Medicine to serve an ageing society. Retired doctors could have a role. PMID- 10669461 TI - Removing barriers to career development in public health. PMID- 10669462 TI - Crinkly toenails. Toenail onychomycosis can cause serious problems. PMID- 10669463 TI - Obituaries PMID- 10669464 TI - Many medical chairs remain unfilled PMID- 10669466 TI - The evolution of british general practice 1850-1948 PMID- 10669465 TI - Cardiovascular medicine: enhanced multimedia CD-ROM PMID- 10669467 TI - Home sweet Home? the impact of poor housing on health PMID- 10669468 TI - Exhibiting the mad psychiatrist PMID- 10669469 TI - The new NHS smoking campaign PMID- 10669470 TI - Consumers' health PMID- 10669472 TI - This tablet may save your life PMID- 10669471 TI - It could be you PMID- 10669473 TI - How best to learn PMID- 10669474 TI - Risk of infection from blood transfusion in UK is negligible PMID- 10669475 TI - False negative result on prenatal Down's screening may damage parental adjustment PMID- 10669476 TI - Food hygiene and declining orofecal infections may explain the allergy and asthma epidemic PMID- 10669477 TI - Lung cancer risk from passive smoking may be overestimated PMID- 10669478 TI - Voluntary sector input benefits patients with psychosocial problems in primary care PMID- 10669479 TI - Improving TB control in prisons may lead to better national programmes PMID- 10669480 TI - Ocular gaze is anchored to the target of an ongoing pointing movement. AB - It is well known that, typically, saccadic eye movements precede goal-directed hand movements to a visual target stimulus. Also pointing in general is more accurate when the pointing target is gazed at. In this study, it is hypothesized that saccades are not only preceding pointing but that gaze also is stabilized during pointing in humans. Subjects, whose eye and pointing movements were recorded, had to make a hand movement and a saccade to a first target. At arm movement peak velocity, when the eyes are usually already fixating the first target, a new target appeared, and subjects had to make a saccade toward it (dynamical trial type). In the statical trial type, a new target was offered when pointing was just completed. In a control experiment, a sequence of two saccades had to be made, with two different interstimulus intervals (ISI), comparable with the ISIs found in the first experiment for dynamic and static trial types. In a third experiment, ocular fixation position and pointing target were dissociated, subjects pointed at not fixated targets. The results showed that latencies of saccades toward the second target were on average 155 ms longer in the dynamic trial types, compared with the static trial types. Saccades evoked during pointing appeared to be delayed with approximately the remaining deceleration time of the pointing movement, resulting in "normal" residual saccadic reaction times (RTs), measured from pointing movement offset to saccade movement onset. In the control experiment, the latency of the second saccade was on average only 29 ms larger when the two targets appeared with a short ISI compared with trials with long ISIs. Therefore the saccadic refractory period cannot be responsible for the substantially bigger delays that were found in the first experiment. The observed saccadic delay during pointing is modulated by the distance between ocular fixation position and pointing target. The largest delays were found when the targets coincided, the smallest delays when they were dissociated. In sum, our results provide evidence for an active saccadic inhibition process, presumably to keep steady ocular fixation at a pointing target and its surroundings. Possible neurophysiological substrates that might underlie the reported phenomena are discussed. PMID- 10669481 TI - Damping actions of the neuromuscular system with inertial loads: soleus muscle of the decerebrate cat. AB - A transient perturbation applied to a limb held in a given posture can induce oscillations. To restore the initial posture, the neuromuscular system must provide damping, which is the dissipation of the mechanical energy imparted by such a perturbation. Despite their importance, damping properties of the neuromuscular system have been poorly characterized. Accordingly, this paper describes the damping characteristics of the neuromuscular system interacting with inertial loads. To quantitatively examine damping, we coupled simulated inertial loads to surgically isolated, reflexively active soleus muscles in decerebrate cats. A simulated force impulse was applied to the load, causing a muscle stretch, which elicited a reflex response. The resulting deviation from the initial position gave rise to oscillations, which decayed progressively. Damping provided by the neuromuscular system was then calculated from the load kinetics. To help interpret our experimental results, we compared our kinetic measurements with those of an analogous linear viscoelastic system and found that the experimental damping properties differed in two respects. First, the amount of damping was greater for large oscillation amplitudes than for small (damping is independent of amplitude in a linear system). Second, plots of force against length during the induced movements showed that damping was greater for shortening than lengthening movements, reflecting greater effective viscosity during shortening. This again is different from the behavior of a linear system, in which damping effects would be symmetrical. This asymmetric and nonlinear damping behavior appears to be related to both the intrinsic nonlinear mechanical properties of the soleus muscle and to stretch reflex properties. The muscle nonlinearities include a change in muscle force-generating capacity induced by forced lengthening, akin to muscle yield, and the nonlinear force-velocity property of muscle, which is different for lengthening versus shortening. Stretch reflex responses are also known to be asymmetric and amplitude dependent. The finding that damping is greater for larger amplitude motion represents a form of automatic gain adjustment to a larger perturbation. In contrast, because of reduced damping at small amplitudes, smaller oscillations would tend to persist, perhaps contributing to normal or "physiological" tremor. This lack of damping for small amplitudes may represent an acceptable compromise for postural regulation in that there is substantial damping for larger movements, where energy dissipation is more critical. Finally, the directional asymmetry in energy dissipation provided by muscle and reflex properties must be reflected in the neural mechanisms for a stable posture. PMID- 10669482 TI - Development of glutamatergic synaptic activity in cultured spinal neurons. AB - The development of glutamatergic synapses involves a sequence of events that are still not well understood. We have studied the time course of the development of glutamatergic synapses in cultured spinal neurons by characterizing spontaneous synaptic currents recorded from cells maintained in vitro for different times. At short times in culture (2 days in vitro; DIV2), spontaneous synaptic activity consisted almost solely of N-methyl-D-aspartate (NMDA) receptor (NMDAR) openings. In contrast, older neurons (DIV5 to DIV8) displayed clear alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA) receptor (AMPAR)-mediated synaptic currents, while the NMDAR-mediated activity remained small. Between 8 and 14 days in vitro there was a large increase in the density of synaptically activated NMDARs, although there was no significant increase in the density of the NMDAR mediated current activated by exogenous glutamate. The results indicate that there is a switch in NMDAR targeting from somatic to synaptic regions during the course of the second in vitro week. Finally, our results support the conclusion that the spontaneous synaptic activity displayed in culture depends on ongoing NMDAR-mediated activity, even when the expression of synaptic NMDARs is low. PMID- 10669483 TI - Inactivation properties of human recombinant class E calcium channels. AB - The electrophysiological and pharmacological properties of alpha(1E)-containing Ca(2+) channels were investigated by using the patch-clamp technique in the whole cell configuration, in HEK 293 cells stably expressing the human alpha(1E) together with alpha(2b) and beta(1b) accessory subunits. These channels had current-voltage (I-V) characteristics resembling those of high-voltage-activated (HVA) Ca(2+) channels (threshold at -30 mV and peak amplitude at +10 mV in 5 mM Ca(2+)). The currents activated and deactivated with a fast rate, in a time- and voltage-dependent manner. No difference was found in their relative permeability to Ca(2+) and Ba(2+). Inorganic Ca(2+) channel blockers (Cd(2+), Ni(2+)) blocked completely and potently the alpha(1E,)/alpha(2b)delta/beta(1b) mediated currents (IC(50) = 4 and 24.6 microM, respectively). alpha(1E)-mediated currents inactivated rapidly and mainly in a non-Ca(2+)-dependent manner, as evidenced by the fact that 1) decreasing extracellular Ca(2+) from 10 to 2 mM and 2) changing the intracellular concentration of the Ca(2+) chelator 1. 2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA), did not affect the inactivation characteristics; 3) there was no clear-cut bell-shaped relationship between test potential and inactivation, as would be expected from a Ca(2+)-dependent event. Although Ba(2+) substitution did not affect the inactivation of alpha(1E) channels, Na(+) substitution revealed a small but significant reduction in the extent and rate of inactivation, suggesting that besides the presence of dominant voltage-dependent inactivation, alpha(1E) channels are also affected by a divalent cation-dependent inactivation process. We have analyzed the Ca(2+) currents produced by a range of imposed action potential-like voltage protocols (APVPs). The amplitude and area of the current were dependent on the duration of the waveform employed and were relatively similar to those described for HVA calcium channels. However, the peak latency resembled that obtained for low voltage-activated (LVA) calcium channels. Short bursts of APVPs applied at 100 Hz produced a depression of the Ca(2+) current amplitude, suggesting an accumulation of inactivation likely to be calcium dependent. The human alpha(1E) gene seems to participate to a Ca(2+) channel type with biophysical and pharmacological properties partly resembling those of LVA and those of HVA channels, with inactivation characteristics more complex than previously believed. PMID- 10669484 TI - NMDA-Receptor-dependent synaptic activation of voltage-dependent calcium channels in basolateral amygdala. AB - Afferent stimulation of pyramidal cells in the basolateral amygdala produced mixed excitatory postsynaptic potentials (EPSPs) mediated by N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors during whole cell current-clamp recordings. In the presence of GABA(A) receptor blockade, the mixed EPSPs recruited a large "all-or-none" depolarizing event. This recruited event was voltage dependent and had a distinct activation threshold. An analogous phenomenon elicited by exogenous glutamate in the presence of tetrodotoxin (TTX) was blocked by Cd(2+), suggesting that the event was a Ca(2+) spike. Selective glutamatergic blockade revealed that these Ca(2+) spikes were recruited readily by single afferent stimulus pulses that elicited NMDA EPSPs. In contrast, non NMDA EPSPs induced by single stimuli failed to elicit the Ca(2+) spike even at maximal stimulus intensities although these non-NMDA EPSPs depolarized the soma more effectively than mixed EPSPs. Elongation of non-NMDA EPSPs by cyclothiazide or brief trains of stimulation were also unable to elicit the Ca(2+) spike. Blockade of K(+) channels with intracellular Cs(+) enabled single non-NMDA EPSPs to activate the Ca(2+) spike. The finding that voltage-dependent calcium channels are activated preferentially by NMDA-receptor-mediated EPSPs provides a mechanism for NMDA-receptor-dependent plasticity independent of Ca(2+) influx through the NMDA receptor. PMID- 10669485 TI - Recurrent excitatory connectivity in the dentate gyrus of kindled and kainic acid treated rats. AB - Repeated seizures induce mossy fiber axon sprouting, which reorganizes synaptic connectivity in the dentate gyrus. To examine the possibility that sprouted mossy fiber axons may form recurrent excitatory circuits, connectivity between granule cells in the dentate gyrus was examined in transverse hippocampal slices from normal rats and epileptic rats that experienced seizures induced by kindling and kainic acid. The experiments were designed to functionally assess seizure-induced development of recurrent circuitry by exploiting information available about the time course of seizure-induced synaptic reorganization in the kindling model and detailed anatomic characterization of sprouted fibers in the kainic acid model. When recurrent inhibitory circuits were blocked by the GABA(A) receptor antagonist bicuculline, focal application of glutamate microdrops at locations in the granule cell layer remote from the recorded granule cell evoked trains of excitatory postsynaptic potentials (EPSPs) and population burst discharges in epileptic rats, which were never observed in slices from normal rats. The EPSPs and burst discharges were blocked by bath application of 1 microM tetrodotoxin and were therefore dependent on network-driven synaptic events. Excitatory connections were detected between blades of the dentate gyrus in hippocampal slices from rats that experienced kainic acid-induced status epilepticus. Trains of EPSPs and burst discharges were also evoked in granule cells from kindled rats obtained after > or = 1 wk of kindled seizures, but were not evoked in slices examined 24 h after a single afterdischarge, before the development of sprouting. Excitatory connectivity between blades of the dentate gyrus was also assessed in slices deafferented by transection of the perforant path, and bathed in artificial cerebrospinal fluid (ACSF) containing bicuculline to block GABA(A) receptor-dependent recurrent inhibitory circuits and 10 mM [Ca(2+)](o) to suppress polysynaptic activity. Low-intensity electrical stimulation of the infrapyramidal blade under these conditions failed to evoke a response in suprapyramidal granule cells from normal rats (n = 15), but in slices from epileptic rats evoked an EPSP at a short latency (2.59 +/- 0.36 ms) in 5 of 18 suprapyramidal granule cells. The results are consistent with formation of monosynaptic excitatory connections between blades of the dentate gyrus. Recurrent excitatory circuits developed in the dentate gyrus of epileptic rats in a time course that corresponded to the development of mossy fiber sprouting and demonstrated patterns of functional connectivity corresponding to anatomic features of the sprouted mossy fiber pathway. PMID- 10669486 TI - Vasopressin increases GABAergic inhibition of rat hypothalamic paraventricular nucleus neurons in vitro. AB - This investigation used an in vitro hypothalamic brain slice preparation and whole cell and perforated-patch recording to examine the response of magnocellular neurons in hypothalamic paraventricular nucleus (PVN) to bath applications of vasopressin (VP; 100-500 nM). In 22/38 cells, responses were characterized by an increase in the frequency of bicuculline-sensitive inhibitory postsynaptic potentials or currents with no detectable influence on excitatory postsynaptic events. Perforated-patch recordings confirmed that VP did not have an effect on intrinsic membrane properties of magnocellular PVN neurons (n = 17). Analysis of intrinsic membrane properties obtained with perforated-patch recording (n = 23) demonstrated that all of nine VP-sensitive neurons showed a rebound depolarization after transient membrane hyperpolarization from rest. By contrast, 12/14 nonresponding neurons displayed a delayed return to resting membrane potentials. Recordings of reversed inhibitory postsynaptic currents with chloride-loaded electrodes showed that responses to VP persisted in media containing glutamate receptor antagonists but were abolished in the presence of tetrodotoxin. In addition, responses were mimicked by vasotocin [Phe(2), Orn(8)], a selective V(1a) receptor agonist, and blocked by [beta-Mercapto-beta, beta cyclopentamethylenepropionyl(1),O-Me-Tyr(2), Arg(8)]-VP (Manning compound), a V(1a)/OT receptor antagonist. Neither [deamino-Cys(1),Val(4),D-Arg(8)]-VP, a selective V(2) receptor agonist, nor oxytocin were effective. Collectively, the results imply that VP acts at V(1a) receptors to excite GABAergic neurons that are presynaptic to a population of magnocellular PVN neurons the identity of which features a unique rebound depolarization. Endogenous sources of VP may be VP-synthesizing neurons in suprachiasmatic nucleus, known to project toward the perinuclear regions of PVN, and/or the magnocellular neurons within PVN. PMID- 10669487 TI - Control of cricket stridulation by a command neuron: efficacy depends on the behavioral state. AB - Crickets use different song patterns for acoustic communication. The stridulatory pattern-generating networks are housed within the thoracic ganglia but are controlled by the brain. This descending control of stridulation was identified by intracellular recordings and stainings of brain neurons. Its impact on the generation of calling song was analyzed both in resting and stridulating crickets and during cercal wind stimulation, which impaired the stridulatory movements and caused transient silencing reactions. A descending interneuron in the brain serves as a command neuron for calling-song stridulation. The neuron has a dorsal soma position, anterior dendritic processes, and an axon that descends in the contralateral connective. The neuron is present in each side of the CNS. It is not activated in resting crickets. Intracellular depolarization of the interneuron so that its spike frequency is increased to 60-80 spikes/s reliably elicits calling-song stridulation. The spike frequency is modulated slightly in the chirp cycle with the maximum activity in phase with each chirp. There is a high positive correlation between the chirp repetition rate and the interneuron's spike frequency. Only a very weak correlation, however, exists between the syllable repetition rate and the interneuron activity. The effectiveness of the command neuron depends on the activity state of the cricket. In resting crickets, experimentally evoked short bursts of action potentials elicit only incomplete calling-song chirps. In crickets that previously had stridulated during the experiment, short elicitation of interneuron activity can trigger sustained calling songs during which the interneuron exhibits a spike frequency of approximately 30 spikes/s. During sustained calling songs, the command neuron activity is necessary to maintain the stridulatory behavior. Inhibition of the interneuron stops stridulation. A transient increase in the spike frequency of the interneuron speeds up the chirp rate and thereby resets the timing of the chirp pattern generator. The interneuron also is excited by cercal wind stimulation. Cercal wind stimulation can impair the pattern of chirp and syllable generation, but these changes are not reflected in the discharge pattern of the command neuron. During wind-evoked silencing reactions, the activity of the calling-song command neuron remains unchanged, but under these conditions, its activity is no longer sufficient to maintain stridulation. Therefore stridulation can be suppressed by cercal inputs from the terminal ganglia without directly inhibiting the descending command activity. PMID- 10669488 TI - Neuropeptide Y(5) receptors reduce synaptic excitation in proximal subiculum, but not epileptiform activity in rat hippocampal slices. AB - Neuropeptide Y (NPY) potently inhibits excitatory synaptic transmission in the hippocampus, acting predominantly via a presynaptic Y(2) receptor. Recent reports that the Y(5) receptor may mediate the anticonvulsant actions of NPY in vivo prompted us to test the hypothesis that Y(5) receptors inhibit synaptic excitation in the hippocampal slice and, furthermore, that they are effective in an in vitro model of anticonvulsant action. Two putative Y(5) receptor-preferring agonists inhibited excitatory postsynaptic currents (EPSCs) evoked by stimulation of stratum radiatum in pyramidal cells. We recorded initially from area CA1 pyramidal cells, but subsequently switched to cells from the subiculum, where a much greater frequency of response was observed to Y(5) agonist application. Both D-Trp(32)NPY (1 microM) and [ahx(8-20)]Pro(34)NPY (3 microM), a centrally truncated, Y(1)/Y(5) agonist we synthesized, inhibited stimulus-evoked EPSCs in subicular pyramidal cells by 44.0 +/- 5.7% and 51.3 +/- 3.5% (mean +/- SE), in 37 and 58% of cells, respectively. By contrast, the less selective centrally truncated agonist, [ahx(8-20)] NPY (1 microM), was more potent (66.4 +/- 4.1% inhibition) and more widely effective, suppressing the EPSC in 86% of subicular neurons. The site of action of all NPY agonists tested was most probably presynaptic, because agonist application caused no changes in postsynaptic membrane properties. The selective Y(1) antagonist, BIBP3226 (1 microM), did not reduce the effect of either more selective agonist, indicating that they activated presynaptic Y(5) receptors. Y(5) receptor-mediated synaptic inhibition was more frequently observed in slices from younger animals, whereas the nonselective agonist appeared equally effective at all ages tested. Because of the similarity with the previously reported actions of Y(2) receptors, we tested the ability of Y(5) receptor agonists to suppress stimulus train-induced bursting (STIB), an in vitro model of ictaform activity, in both area CA3 and the subiculum. Neither [ahx(8-20)]Pro(34)NPY nor D-Trp(32)NPY were significantly effective in suppressing or shortening STIB-induced afterdischarge, with <20% of slices responding to these agonists in recordings from CA3 and none in subiculum. By contrast, 1 microM each of [ahx(8-20)]NPY, the Y(2) agonist, [ahx(5-24)]NPY, and particularly NPY itself suppressed the afterdischarge in area CA3 and the subiculum, as reported earlier. We conclude that Y(5) receptors appear to regulate excitability to some degree in the subiculum of young rats, but their contribution is relatively small compared with those of Y(2) receptors, declines with age, and is insufficient to block or significantly attenuate STIB-induced afterdischarges. PMID- 10669489 TI - Na(+) and K(+) concentrations, extra- and intracellular voltages, and the effect of TTX in hypoxic rat hippocampal slices. AB - Severe hypoxia causes rapid depolarization of CA1 neurons and glial cells that resembles spreading depression (SD). In brain slices in vitro, the SD-like depolarization and the associated irreversible loss of function can be postponed, but not prevented, by blockade of Na(+) currents by tetrodotoxin (TTX). To investigate the role of Na(+) flux, we made recordings from the CA1 region in hippocampal slices in the presence and absence of TTX. We measured membrane changes in single CA1 pyramidal neurons simultaneously with extracellular DC potential (V(o)) and either extracellular [K(+)] or [Na(+)]; alternatively, we simultaneously recorded [Na(+)](o), [K(+)](o), and V(o). Confirming previous reports, early during hypoxia, before SD onset, [K(+)](o) began to rise, whereas [Na(+)](o) still remained normal and V(o) showed a slight, gradual, negative shift; neurons first hyperpolarized and then began to gradually depolarize. The SD-like abrupt negative DeltaV(o) corresponded to a near complete depolarization of pyramidal neurons and an 89% decrease in input resistance. [K(+)](o) increased by 47 mM and [Na(+)](o) dropped by 91 mM. Changes in intracellular Na(+) and K(+) concentrations, estimated on the basis of the measured extracellular ion levels and the relative volume fractions of the neuronal, glial, and extracellular compartment, were much more moderate. Because [Na(+)](o) dropped more than [K(+)](o) increased, simple exchange of Na(+) for K(+) cannot account for these ionic changes. The apparent imbalance of charge could be made up by Cl(-) influx into neurons paralleling Na(+) flux and release of Mg(2+) from cells. The hypoxia induced changes in interneurons resembled those observed in pyramidal neurons. Astrocytes responded with an initial slow depolarization as [K(+)](o) rose. It was followed by a rapid but incomplete depolarization as soon as SD occurred, which could be accounted for by the reduced ratio, [K(+)](i)/[K(+)](o). TTX (1 microM) markedly postponed SD, but the SD-related changes in [K(+)](o) and [Na(+)](o) were only reduced by 23 and 12%, respectively. In TTX-treated pyramidal neurons, the delayed SD-like depolarization took off from a more positive level, but the final depolarized intracellular potential and input resistance were not different from control. We conclude that TTX-sensitive channels mediate only a fraction of the Na(+) influx, and that some of the K(+) is released in exchange for Na(+). Even though TTX-sensitive Na(+) currents are not essential for the self-regenerative membrane changes during hypoxic SD, in control solutions their activation may trigger the transition from gradual to rapid depolarization of neurons, thereby synchronizing the SD-like event. PMID- 10669490 TI - Low-voltage-activated calcium current does not regulate the firing behavior in paired mechanosensory neurons with different adaptation properties. AB - Low-voltage-activated Ca(2+) currents (LVA-I(Ca)) are believed to perform several roles in neurons such as lowering the threshold for action potentials, promoting burst firing and oscillatory behavior, and enhancing synaptic excitation. They also may allow rapid increases in intracellular Ca(2+) concentration. We discovered LVA-I(Ca) in both members of paired mechanoreceptor neurons in a spider, where one neuron adapts rapidly (Type A) and the other slowly (Type B) in response to a step stimulus. To learn if I(Ca) contributed to the difference in adaptation behavior, we studied the kinetics of I(Ca) from isolated somata under single-electrode voltage-clamp and tested its physiological function under current clamp. LVA-I(Ca) was large enough to fire single action potentials when all other voltage-activated currents were blocked, but we found no evidence that it regulated firing behavior. LVA-I(Ca) did not lower the action potential threshold or affect firing frequency. Previous experiments have failed to find Ca(2+)-activated K(+) current (I(K(Ca))) in the somata of these neurons, so it is also unlikely that LVA-I(Ca) interacts with I(K(Ca)) to produce oscillatory behavior. We conclude that LVA-Ca(2+) channels in the somata, and possible in the dendrites, of these neurons open in response to the depolarization caused by receptor current and by the voltage-activated Na(+) current (I(Na)) that produces action potential(s). However, the role of the increased intracellular Ca(2+) concentration in neuronal function remains enigmatic. PMID- 10669491 TI - Relationships between odor-elicited oscillations in the salamander olfactory epithelium and olfactory bulb. AB - Oscillations in neuronal population activity, or the synchronous neuronal spiking that underlies them, are thought to play a functional role in sensory processing in the CNS. In the olfactory system, stimulus-induced oscillations are observed both in central processing areas and in the peripheral receptor epithelium. To examine the relationship between these peripheral and central oscillations, we recorded local field potentials simultaneously from the olfactory epithelium and olfactory bulb in tiger salamanders (Ambystoma tigrinum). Stimulus-induced oscillations recorded at these two sites were matched in frequency and slowed concurrently over the time course of the response, suggesting that the oscillations share a common source or are modulated together. Both the power and duration of oscillations increased over a range of amyl acetate concentrations from 2.5 x 10(-2) to 1 x 10(-1) dilution of saturated vapor, but peak frequency was not affected. The frequency of the oscillation did vary with different odorant compounds in both olfactory epithelium and bulb (OE and OB): amyl acetate, ethyl fenchol and d-carvone elicited oscillations of significantly different frequencies, and there was no difference in OE and OB oscillation frequencies. No change in the power or frequency of OE oscillations was observed after sectioning the olfactory nerve, indicating that the OE oscillations have a peripheral source. Finally, application of 1.0 and 10 microM tetrodotoxin to the epithelium blocked OE oscillations in a dose-dependent and reversible manner, suggesting that peripheral olfactory oscillations are related to receptor neuron spiking. PMID- 10669492 TI - Prototypical imidazoline-1 receptor ligand moxonidine activates alpha2 adrenoceptors in bulbospinal neurons of the RVL. AB - Moxonidine is an antihypertensive drug that lowers sympathetic vasomotor tone by stimulating either alpha2-adrenergic (alpha2-AR) or imidazoline I1 receptors within the rostral ventrolateral medulla (RVL). In this study, we investigated the effects of moxonidine (10 microM) on RVL neurons in brain stem slices of neonatal rats. We recorded mainly from retrogradely labeled RVL bulbospinal neurons (putative presympathetic neurons) except for some extracellular recordings. Prazosin was used to block alpha1-adrenoceptors. Moxonidine inhibited the extracellularly recorded discharges of all spontaneously active RVL neurons tested (bulbospinal and unidentified). This effect was reversed or blocked by the selective alpha2-AR antagonist SKF 86466 (10 microM). In contrast, the I1 imidazoline ligand AGN 192403 (10 microM) had no effect on the spontaneous activity. In whole cell recordings (holding potential -70 mV), moxonidine produced a small and variable outward current (mean 7 pA). This current was observed in both tyrosine hydroxylase-immunoreactive and other bulbospinal neurons and was blocked by SKF 86466. Excitatory postsynaptic currents (EPSCs) evoked by focal electrical stimulation were isolated by incubation with gabazine and strychnine, and inhibitory postsynaptic currents (IPSCs) were isolated with 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX). Moxonidine reduced the amplitude of the evoked EPSCs (EC(50) = 1 microM; 53% inhibition at 10 microM) but not their decay time constant (5.6 ms). The effect of moxonidine on EPSCs persisted in barium (300 microM) and was reduced approximately 80% by SKF 86466. Moxonidine also reduced the amplitude of evoked IPSCs by 63%. In conclusion, moxonidine inhibits putative RVL presympathetic neurons both presynaptically and postsynaptically. All observed effects in the present study are consistent with an alpha2-AR agonist activity of moxonidine. PMID- 10669493 TI - Effects of attention on MT and MST neuronal activity during pursuit initiation. AB - The responses of neurons in monkey extrastriate areas MT (middle temporal) and MST (medial superior temporal), and the initial metrics of saccadic and pursuit eye movements, have previously been shown to be better predicted by vector averaging or winner-take-all models depending on the stimulus conditions. To investigate the potential influences of attention on the neuronal activity, we measured the responses of single MT and MST neurons under identical stimulus conditions when one of two moving stimuli was the target for a pursuit eye movement. We found the greatest attentional modulation across neurons when two stimuli moved through the receptive field (RF) of the neuron and the stimulus motion was initiated at least 450 ms before reaching the center of the RF. These conditions were the same as those in which a winner-take-all model better predicted both the eye movements and the underlying neuronal activity. The modulation was almost always an increase of activity, and it was about equally frequent in MT and MST. A modulation of >50% was observed in approximately 41% of MT neurons and 27% of MST neurons. Responses to all directions of motion were modulated so that the direction tuning curves in the attended and unattended conditions were similar. Changes in the background activity with target selection were small and unlikely to account for the observed attentional modulation. In contrast, there was little change in the neuronal response with attention when the stimulus reached the RF center 150 ms after motion onset, which was also the condition in which the vector average model better predicted the initial eye movements and the activity of the neurons. These results are consistent with a competition model of attention in which top-down attention acts on the activity of one of two competing populations of neurons activated by the bottom-up input from peripheral stimuli. They suggest that there is a minimal separation of the populations necessary before attention can act on one population, similar to that required to produce a winner-take-all mode of behavior in pursuit initiation. The present experiments also suggest that it takes several hundred milliseconds to develop this top-down attention effect. PMID- 10669494 TI - Effect of high Ba(2+) on norepinephrine-induced inhibition of N-type calcium current in bullfrog sympathetic neurons. AB - The voltage-dependent inhibition of N-type calcium current by neurotransmitters is the best-understood example of neuronal calcium channel inhibition. One of the mechanisms by which this pathway is thought to inhibit the calcium current is by reducing the permeation of divalent cations through the channel. In this study one prediction of this hypothesis was examined, that high concentrations of divalent cations reduce the maximum neurotransmitter-induced inhibition. Norepinephrine (NE)-induced inhibition was compared in external solutions containing either 2 or 100 mM Ba(2+). Initially, NE dose-response curves were generated by averaging data from many neurons, and it was found that the relationship was right shifted in the high-Ba(2+) external solution without an effect on maximum inhibition. The IC(50) was 0.6 and 3 microM in 2 and 100 mM Ba(2+), respectively. This shift was verified by comparing the effect of NE on single neurons exposed to both 2 and 100 mM Ba(2+). The inhibition induced by 1 microM NE was reduced in 100 mM Ba(2+) compared with that in 2 mM Ba(2+). However, the response to 100 microM NE was identical between high and low Ba(2+). Thus, divalent cations appear to act as a competitive inhibitor of NE binding, which likely results from these ions' interacting with negatively charged amino acids that are important for catecholamine binding to adrenergic receptors. Because the maximum inhibition induced by NE was similar in low and high Ba(2+), the effect of inhibition on single N-type calcium channels was not altered by the divalent cation concentration. PMID- 10669495 TI - Conditioned eyeblink response consists of two distinct components. AB - The aim of these experiments was to obtain a detailed knowledge of how the orbicularis oculi muscle is activated during the execution of a conditioned eyeblink response (CR). This is the first critical step to understand the underlying neural mechanisms involved in the control of the CR. Decerebrate ferrets were trained in a classical conditioning paradigm. The conditioned stimulus (CS) was a train of electrical stimuli (15 pulses, 50 Hz, 1 mA) applied to the forelimb, and the unconditioned stimulus (US) was a train of electrical stimuli (3 pulses, 50 Hz, 3-4 mA) to the periorbital region. The CRs were studied by recording electromyograms (EMGs) from the orbicularis oculi muscle. The eyeblink CR in all animals showed a similar topography with at least two different components, CR1 and CR2, which were expressed at different rates. CR1 appeared first during acquisition, had a shorter onset latency, and was more phasic and more resistant to extinction than CR2. A marked pause in the muscle activity separated the two components. To control that the two-component CR were not species, paradigm or preparation specific, awake rabbits were trained with a tone CS (300 ms, 4 kHz, 64 dB) and a train of periorbital stimuli as US (3 pulses, 50 Hz, 3 mA). CR1 and CR2 were present in the rabbit eyeblink CR. The cerebellum is implicated in the control of CRs and to study whether separate neural pathways were responsible for CR1 and CR2, direct brachium pontis stimulation was used to replace the forelimb CS. CR1 and CR2 were present in the CR elicited by the brachium pontis CS. The presence of CR1 and CR2 after a unilateral lesion of the brachium conjunctivum shows that output from the contralateral cerebellar hemisphere was not the cause for any of the components. Other mechanisms that might be involved in the separation of the CR into two components are discussed. The results show that the eyeblink CR consists of at least two components, CR1 and CR2, which most likely originate either as a direct central command from the cerebellum or in the output pathway before the facial nucleus. PMID- 10669496 TI - Intrinsic dynamics in neuronal networks. I. Theory. AB - Many networks in the mammalian nervous system remain active in the absence of stimuli. This activity falls into two main patterns: steady firing at low rates and rhythmic bursting. How are these firing patterns generated? Specifically, how do dynamic interactions between excitatory and inhibitory neurons produce these firing patterns, and how do networks switch from one firing pattern to the other? We investigated these questions theoretically by examining the intrinsic dynamics of large networks of neurons. Using both a semianalytic model based on mean firing rate dynamics and simulations with large neuronal networks, we found that the dynamics, and thus the firing patterns, are controlled largely by one parameter, the fraction of endogenously active cells. When no endogenously active cells are present, networks are either silent or fire at a high rate; as the number of endogenously active cells increases, there is a transition to bursting; and, with a further increase, there is a second transition to steady firing at a low rate. A secondary role is played by network connectivity, which determines whether activity occurs at a constant mean firing rate or oscillates around that mean. These conclusions require only conventional assumptions: excitatory input to a neuron increases its firing rate, inhibitory input decreases it, and neurons exhibit spike-frequency adaptation. These conclusions also lead to two experimentally testable predictions: 1) isolated networks that fire at low rates must contain endogenously active cells and 2) a reduction in the fraction of endogenously active cells in such networks must lead to bursting. PMID- 10669497 TI - Intrinsic dynamics in neuronal networks. II. experiment. AB - Neurons in many regions of the mammalian CNS remain active in the absence of stimuli. This activity falls into two main patterns: steady firing at low rates and rhythmic bursting. How these firing patterns are maintained in the presence of powerful recurrent excitation, and how networks switch between them, is not well understood. In the previous paper, we addressed these issues theoretically; in this paper we address them experimentally. We found in both studies that a key parameter in controlling firing patterns is the fraction of endogenously active cells. The theoretical analysis indicated that steady firing rates are possible only when the fraction of endogenously active cells is above some threshold, that there is a transition to bursting when it falls below that threshold, and that networks becomes silent when the fraction drops to zero. Experimentally, we found that all steadily firing cultures contain endogenously active cells, and that reducing the fraction of such cells in steadily firing cultures causes a transition to bursting. The latter finding implies indirectly that the elimination of endogenously active cells would cause a permanent drop to zero firing rate. The experiments described here thus corroborate the theoretical analysis. PMID- 10669498 TI - Kinetic and frequency-domain properties of reflex and conditioned eyelid responses in the rabbit. AB - Eyelid position and the electromyographic activity of the orbicularis oculi muscle were recorded unilaterally in rabbits during reflex and conditioned blinks. Air-puff-evoked blinks consisted of a fast downward phase followed sometimes by successive downward sags. The reopening phase had a much longer duration and slower peak velocity. Onset latency, maximum amplitude, peak velocity, and rise time of reflex blinks depended on the intensity and duration of the air puff-evoking stimulus. A flashlight focused on the eye also evoked reflex blinks, but not flashes of light, or tones. Both delayed and trace classical conditioning paradigms were used. For delayed conditioning, animals were presented with a 350-ms, 90-dB, 600-Hz tone, as conditioned stimulus (CS). For trace conditioning, animals were presented with a 10-ms, 1-k/cm(2) air puff, as CS. The unconditioned stimulus (US) consisted of a 100-ms, 3-k/cm(2) air puff. The stimulus interval between CS and US onsets was 250 ms. Conditioned responses (CRs) to tones were composed of downward sags that increased in number through the successive conditioning sessions. The onset latency of the CR decreased across conditioning at the same time as its maximum amplitude and its peak velocity increased, but the time-to-peak of the CR remained unaltered. The topography of CRs evoked by short, weak air puffs as the CS showed three different components: the alpha response to the CS, the CR, and the reflex response to the US. Through conditioning, CRs showed a decrease in onset latency, and an increase in maximum amplitude and peak velocity. The time-to-peak of the CR remained unchanged. A power spectrum analysis of reflex and conditioned blink acceleration profiles showed a significant approximately 8-Hz oscillation within a broadband of frequencies between 4 and 15 Hz. Nose and mandible movements presented power spectrum profiles different from those characterizing reflex and conditioned blinks. It is concluded that eyelid reflex responses in the rabbit present significant differences from CRs in their profiles and metric properties, suggesting different neural origins, but that a common approximately 8-Hz neural oscillator underlies lid motor performance. According to available data, the frequency of this putative oscillator seems to be related to the species size. PMID- 10669499 TI - Activity of reticulospinal neurons during locomotion in the freely behaving lamprey. AB - The reticulospinal (RS) system is the main descending system transmitting commands from the brain to the spinal cord in the lamprey. It is responsible for initiation of locomotion, steering, and equilibrium control. In the present study, we characterize the commands that are sent by the brain to the spinal cord in intact animals via the reticulospinal pathways during locomotion. We have developed a method for recording the activity of larger RS axons in the spinal cord in freely behaving lampreys by means of chronically implanted macroelectrodes. In this paper, the mass activity in the right and left RS pathways is described and the correlations of this activity with different aspects of locomotion are discussed. In quiescent animals, the RS neurons had a low level of activity. A mild activation of RS neurons occurred in response to different sensory stimuli. Unilateral eye illumination evoked activation of the ipsilateral RS neurons. Unilateral illumination of the tail dermal photoreceptors evoked bilateral activation of RS neurons. Water vibration also evoked bilateral activation of RS neurons. Roll tilt evoked activation of the contralateral RS neurons. With longer or more intense sensory stimulation of any modality and laterality, a sharp, massive bilateral activation of the RS system occurred, and the animal started to swim. This high activity of RS neurons and swimming could last for many seconds after termination of the stimulus. There was a positive correlation between the level of activity of RS system and the intensity of locomotion. An asymmetry in the mass activity on the left and right sides occurred during lateral turns with a 30% prevalence (on average) for the ipsilateral side. Rhythmic modulation of the activity in RS pathways, related to the locomotor cycle, often was observed, with its peak coinciding with the electromyographic (EMG) burst in the ipsilateral rostral myotomes. The pattern of vestibular response of RS neurons observed in the quiescent state, that is, activation with contralateral roll tilt, was preserved during locomotion. In addition, an inhibition of their activity with ipsilateral tilt was clearly seen. In the cases when the activity of individual neurons could be traced during swimming, it was found that rhythmic modulation of their firing rate was superimposed on their tonic firing or on their vestibular responses. In conclusion, different aspects of locomotor activity-initiation and termination, vigor of locomotion, steering and equilibrium control-are well reflected in the mass activity of the larger RS neurons. PMID- 10669500 TI - Responses of reticulospinal neurons in intact lamprey to vestibular and visual inputs. AB - A lamprey maintains the dorsal-side-up orientation due to the activity of postural control system driven by vestibular input. Visual input can affect the body orientation: illumination of one eye evokes ipsilateral roll tilt. An important element of the postural network is the reticulospinal (RS) neurons transmitting commands from the brain stem to the spinal cord. Here we describe responses to vestibular and visual stimuli in RS neurons of the intact lamprey. We recorded activity from the axons of larger RS neurons with six extracellular electrodes chronically implanted on the surface of the spinal cord. From these multielectrode recordings of mass activity, discharges in individual axons were extracted by means of a spike-sorting program, and the axon position in the spinal cord and its conduction velocity were determined. Vestibular stimulation was performed by rotating the animal around its longitudinal axis in steps of 45 degrees through 360 degrees. Nonpatterned visual stimulation was performed by unilateral eye illumination. All RS neurons were classified into two groups depending on their pattern of response to vestibular and visual stimuli; the groups also differed in the axon position in the spinal cord and its conduction velocity. Each group consisted of two symmetrical, left and right, subgroups. In group 1 neurons, rotation of the animal evoked both dynamic and static responses; these responses were much larger when rotation was directed toward the contralateral labyrinth, and the dynamic responses to stepwise rotation occurred at any initial orientation of the animal, but they were more pronounced within the angular zone of 0-135 degrees. The zone of static responses approximately coincided with the zone of pronounced dynamic responses. The group 1 neurons received excitatory input from the ipsilateral eye and inhibitory input from the contralateral eye. When vestibular stimulation was combined with illumination of the ipsilateral eye, both dynamic and static vestibular responses were augmented. Contralateral eye illumination caused a decrease of both types of responses. Group 2 neurons responded dynamically to rotation in both directions throughout 360 degrees. They received excitatory inputs from both eyes. Axons of the group 2 neurons had higher conduction velocity and were located more medially in the spinal cord as compared with the group 1 neurons. We suggest that the reticulospinal neurons of group 1 constitute an essential part of the postural network in the lamprey. They transmit orientation-dependent command signals to the spinal cord causing postural corrections. The role of these neurons is discussed in relation to the model of the roll control system formulated in our previous studies. PMID- 10669501 TI - 2-Deoxyglucose-induced long-term potentiation of monosynaptic IPSPs in CA1 hippocampal neurons. AB - In previous experiments on excitatory synaptic transmission in CA1, temporary (10 20 min) replacement of glucose with 10 mM 2-deoxyglucose (2-DG) consistently caused a marked and very sustained potentiation (2-DG LTP). To find out whether 2 DG has a similar effect on inhibitory synapses, we recorded pharmacologically isolated mononosynaptic inhibitory postsynaptic potentials (IPSPs; under current clamp) and inhibitory postsynaptic currents (IPSCs; under voltage clamp); 2-DG was applied both in the presence and the absence of antagonists of N-methyl-D aspartate (NMDA). In spite of sharply varied results (some neurons showing large potentiation, lasting for >1 h, and many little or none), overall there was a significant and similar potentiation of IPSP conductance, both for the early (at approximately 30 ms) and later (at approximately 140 ms) components of IPSPs or IPSCs: by 35.1 +/- 10.25% (mean +/- SE; for n = 24, P = 0.0023) and 36.5 +/- 16.3% (for n = 19, P = 0.038), respectively. The similar potentiation of the early and late IPSP points to a presynaptic mechanism of LTP. Overall, the LTP was statistically significant only when 2-DG was applied in the absence of glutamate antagonists. Tetanic stimulations (in presence or absence of glutamate antagonists) only depressed IPSPs (by half). In conclusion, although smaller and more variable, 2-DG-induced LTP of inhibitory synapses appears to be broadly similar to the 2-DG-induced LTP of excitatory postsynaptic potentials previously observed in CA1. PMID- 10669502 TI - Sacrocaudal afferents induce rhythmic efferent bursting in isolated spinal cords of neonatal rats. AB - The ability of mammalian spinal cords to generate rhythmic motor behavior in nonlimb moving segments was examined in isolated spinal cords of neonatal rats. Stimulation of sacrocaudal afferents (SCA) induced alternating left-right bursts in lumbosacral efferents and in tail muscles. On each side of the tail, flexors, extensors, and abductors were coactive during each cycle of activity. This rhythm originated mainly in the sacrocaudal region because it persisted in sacrocaudal segments after surgical removal of the thoracolumbar cord. Sacrocaudal commissural pathways were sufficient to maintain the left-right alternation of lumbar efferent bursts, because their timing was unaltered after a complete thoracolumbar hemisection. The lumbar rhythm originated in part from sacrocaudal activity ascending in lateral and ventrolateral funiculi, because efferent bursts in rostral lumbar segments were nearly abolished on a particular side by lesions of the lateral quadrant of the cord at the L(4)-L(5) junction. Intracellular recordings from S(2)-S(3) motoneurons, obtained during the rhythm, revealed the presence of phasic oscillations of membrane potential superimposed on a tonic depolarization. Bursts of spikes occurred on the depolarizing phases of the oscillation. Between these bursts the membrane input conductance increased, and hyperpolarizing drive potentials were revealed. The inhibitory drive and the decreased input resistance coincided with contralateral efferent bursts, suggesting that crossed pathways controlled it. Our studies indicate that pattern generators are not restricted to limb-moving spinal segments and suggest that regional specializations of pattern-generating circuitry and their associated interneurons are responsible for the different motor patterns produced by the mammalian spinal cord. PMID- 10669503 TI - Impairments in prehension produced by early postnatal sensory motor cortex activity blockade. AB - This study examined the effects of blocking neural activity in sensory motor cortex during early postnatal development on prehension. We infused muscimol, either unilaterally or bilaterally, into the sensory motor cortex of cats to block activity continuously between postnatal weeks 3-7. After stopping infusion, we trained animals to reach and grasp a cube of meat and tested behavior thereafter. Animals that had not received muscimol infusion (unilateral saline infusion; age-matched) reached for the meat accurately with small end-point errors. They grasped the meat using coordinated digit flexion followed by forearm supination on 82.7% of trials. Performance using either limb did not differ significantly. In animals receiving unilateral muscimol infusion, reaching and grasping using the limb ipsilateral to the infusion were similar to controls. The limb contralateral to infusion showed significant increases in systematic and variable reaching end-point errors, often requiring subsequent corrective movements to contact the meat. Grasping occurred on only 14.8% of trials, replaced on most trials by raking without distal movements. Compensatory adjustments in reach length and angle, to maintain end-point accuracy as movements were started from a more lateral position, were less effective using the contralateral limb than ipsilateral limb. With bilateral inactivations, the form of reaching and grasping impairments was identical to that produced by unilateral inactivation, but the magnitude of the reaching impairments was less. We discuss these results in terms of the differential effects of unilateral and bilateral inactivation on corticospinal tract development. We also investigated the degree to which these prehension impairments after unilateral blockade reflect control by each hemisphere. In animals that had received unilateral blockade between postnatal weeks (PWs) 3 and 7, we silenced on-going activity (after PW 11) during task performance using continuous muscimol infusion. We inactivated the right (previously active) and then the left (previously silenced) sensory motor cortex. Inactivation of the ipsilateral (right) sensory motor cortex produced a further increase in systematic error and less frequent normal grasping. Reinactivation of the contralateral (left) cortex produced larger increases in reaching and grasping impairments than those produced by ipsilateral inactivation. This suggests that the impaired limb receives bilateral sensory motor cortex control but that control by the contralateral (initially silenced) cortex predominates. Our data are consistent with the hypothesis that the normal development of skilled motor behavior requires activity in sensory motor cortex during early postnatal life. PMID- 10669504 TI - Directionality derived from pinna-cue spectral notches in cat dorsal cochlear nucleus. AB - We tested two hypotheses to determine whether dorsal cochlear nucleus (DCN) neurons are specialized to derive directionality from spectral notches: DCN neurons exhibit greater spectral-dependent directionality than ventral cochlear nucleus (VCN) neurons, and spectral-dependent directionality depends on response minima (nulls) produced by coincidence of best frequency (BF) and spectral-notch center frequency. Single-unit responses to 50-ms noise and tone bursts were recorded in barbiturate-anesthetized cats (BFs: 4-37 kHz). Units were classified using BF tone poststimulus time histograms. Pauser, onset-G (type II interneurons), and some chopper units were recorded from the DCN. Primary-like, onset-CIL (onset other than onset-G), and most choppers in the sample were recorded from the VCN. Many pauser and onset-G units were highly directional to noise. Chopper, onset-CIL, and primary-like units (collectively referred to as C O-P units) were not. The difference in directionality depends on a monaural mechanism as pausers were more directional to monaural noise than C-O-P units. Contralateral inhibition produced a small increase in pauser directionality to noise simulation but had no effect on directionality of C-O-P units. Pauser and C O-P units exhibited similar low directionality to BF tone, showing that the difference in noise directionality between groups depends on spectral cues. These results show that spectral-dependent directionality is a DCN specialization. Azimuth functions of highly directional units exhibited response nulls, and there was a linear relationship between BFs in the range of 8-13 kHz and azimuthal locations of nulls. This relationship parallels the known spatial distribution of spectral-notch center frequencies on the horizontal plane. Furthermore spatial receptive fields of pausers show response nulls that follow the expected diagonal trajectory of the spectral notch in this frequency range. These results show that DCN spectral-dependent directionality depends on response nulls produced by coincidence of unit BF and spectral-notch center-frequency. PMID- 10669505 TI - Pharmacological evidence of inhibitory and disinhibitory neuronal circuits in dorsal cochlear nucleus. AB - The dorsal cochlear nucleus (DCN) is rich in both glycine and GABA inhibitory neurotransmitter systems, and the response properties of its principal cells (pyramidal and giant cells) are strongly shaped by inhibitory inputs. For example, DCN principal cells often display highly nonmonotonic (so-called type IV) input-output functions in response to best-frequency (BF) tones. In this study, the inhibitory inputs onto the principal cell types and onto response types of known inhibitory interneurons were compared before and during iontophoretic application of the glycine- and GABA(A)-receptor antagonists, strychnine and bicuculline. Strychnine eliminates the central (on-BF) inhibitory area in type IV units, resulting in monotonic BF rate-level curves. Unexpectedly, bicuculline primarily enhances inhibition in principal-cell types; for example, type IV units are inhibited at lower sound levels in the presence of bicuculline. Principal cell types with weaker inhibitory inputs (type IV-T and type III units) are more strongly inhibited in the presence of bicuculline and usually are converted into type IV units. This enhancement of on-BF inhibition by bicuculline suggests a disinhibitory process involving GABA(A) action on a non-GABA(A)ergic inhibitory pathway. This latter pathway is probably glycinergic and involves type II units (deep-layer vertical cells) and/or complex-spiking units (superficial cartwheel cells) because both of these unit types are disinhibited by bicuculline. One intrinsic GABA(A) source could be the superficial stellate cells in DCN because bicuculline partly blocks the inhibition evoked by somatosensory stimulated activation of the superficial granule-cell circuitry in DCN. Taken together, the results suggest that glycinergic circuits mediate directly the inhibition of DCN principal cells, but that GABA(A)ergic circuits modulate the strength of the inhibition. PMID- 10669506 TI - Contextual influence on orientation discrimination of humans and responses of neurons in V1 of alert monkeys. AB - We studied the effects of various patterns as contextual stimuli on human orientation discrimination, and on responses of neurons in V1 of alert monkeys. When a target line is presented along with various contextual stimuli (masks), human orientation discrimination is impaired. For most V1 neurons, responses elicited by a line in the receptive field (RF) center are suppressed by these contextual patterns. Orientation discrimination thresholds of human observers are elevated slightly when the target line is surrounded by orthogonal lines. For randomly oriented lines, thresholds are elevated further and even more so for lines parallel to the target. Correspondingly, responses of most V1 neurons to a line are suppressed. Although contextual lines inhibit the amplitude of orientation tuning functions of most V1 neurons, they do not systematically alter the tuning width. Elevation of human orientation discrimination thresholds decreases with increasing curvature of masking lines, so does the inhibition of V1 neuronal responses. A mask made of straight lines yields the strongest interference with human orientation discrimination and produces the strongest suppression of neuronal responses. Elevation of human orientation discrimination thresholds is highest when a mask covers only the immediate vicinity of the target line. Increasing the masking area results in less interference. On the contrary, suppression of neuronal responses in V1 increases with increasing mask size. Our data imply that contextual interference observed in human orientation discrimination is in part directly related to contextual inhibition of neuronal activity in V1. However, the finding that interference with orientation discrimination is weaker for larger masks suggests a figure-ground segregation process that is not located in V1. PMID- 10669507 TI - Silent NMDA receptor-mediated synapses are developmentally regulated in the dorsal horn of the rat spinal cord. AB - In vitro whole cell patch-clamp recording techniques were utilized to study silent pure-N-methyl-D-aspartate (NMDA) receptor-mediated synaptic responses in lamina II (substantia gelatinosa, SG) and lamina III of the spinal dorsal horn. To clarify whether these synapses are present in the adult and contribute to neuropathic pain, transverse lumbar spinal cord slices were prepared from neonatal, naive adult and adult sciatic nerve transected rats. In neonatal rats, pure-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) were elicited in SG neurons either by focal intraspinal stimulation (n = 15 of 20 neurons) or focal stimulation of the dorsal root (n = 2 of 7 neurons). In contrast, in slices from naive adult rats, no silent pure-NMDA EPSCs were recorded in SG neurons following focal intraspinal stimulation (n = 27), and only one pure-NMDA EPSC was observed in lamina III (n = 23). Furthermore, in rats with chronic sciatic nerve transection, pure-NMDA EPSCs were elicited by focal intraspinal stimulation in only 2 of 45 SG neurons. Although a large increase in Abeta fiber evoked mixed alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and NMDA receptor-mediated synapses was detected after sciatic nerve injury, Abeta fiber-mediated pure-NMDA EPSCs were not evoked in SG neurons by dorsal root stimulation. Pure-NMDA receptor-mediated EPSCs are therefore a transient, developmentally regulated phenomenon, and, although they may have a role in synaptic refinement in the immature dorsal horn, they are unlikely to be involved in receptive field plasticity in the adult. PMID- 10669508 TI - Effects of intracolonic opioid receptor agonists on polymodal pelvic nerve afferent fibers in the rat. AB - We studied the effects of intracolonic administration of opioid receptor agonists (ORAs) on responses of pelvic nerve afferent fibers to colorectal distension (CRD) and heat. Single-fiber recordings were made from the decentralized S1 dorsal rootlet in the rat. An approximately 7-cm length of descending colon was isolated in situ to permit intracolonic perfusion with Krebs solution, which, when the outflow was clamped, was used to distend the colon. Responses to noxious CRD (40 mmHg, 30 s) were tested after intracolonic instillation of mu-, delta- or kappa-ORAs. Intracolonic administration of the kappa-ORAs EMD 61,753 (n = 5/12) and U62,066 (n = 8/11), but not either the mu-ORA fentanyl or the delta-ORA SNC 80, concentration-dependently inhibited responses of afferent fibers. For fibers unaffected by intracolonic administration of EMD 61,753 or U62,066, intra arterial administration of kappa-ORAs was effective. Forty-one of 54 mechanosensitive fibers also responded to intracolonic instillation of heated Krebs solution (50 degrees C). Intra-arterial injection of fentanyl or SNC-80 did not attenuate responses to heat. Either intracolonic or intra-arterial administration of EMD 61,753 or U62, 066, however, inhibited afferent fiber responses to heat. These results document that mechanical and thermal sensitivity of polymodal pelvic nerve afferent fibers innervating the rat colon can be inhibited peripherally by intracolonic instillation of kappa-ORAs. PMID- 10669509 TI - Precision of the pacemaker nucleus in a weakly electric fish: network versus cellular influences. AB - We investigated the relative influence of cellular and network properties on the extreme spike timing precision observed in the medullary pacemaker nucleus (Pn) of the weakly electric fish Apteronotus leptorhynchus. Of all known biological rhythms, the electric organ discharge of this and related species is the most temporally precise, with a coefficient of variation (CV = standard deviation/mean period) of 2 x 10(-4) and standard deviation (SD) of 0.12-1.0 micros. The timing of the electric organ discharge is commanded by neurons of the Pn, individual cells of which we show in an in vitro preparation to have only a slightly lesser degree of precision. Among the 100-150 Pn neurons, dye injection into a pacemaker cell resulted in dye coupling in one to five other pacemaker cells and one to three relay cells, consistent with previous results. Relay cell fills, however, showed profuse dendrites and contacts never seen before: relay cell dendrites dye coupled to one to seven pacemaker and one to seven relay cells. Moderate (0.1-10 nA) intracellular current injection had no effect on a neuron's spiking period, and only slightly modulated its spike amplitude, but could reset the spike phase. In contrast, massive hyperpolarizing current injections (15-25 nA) could force the cell to skip spikes. The relative timing of subthreshold and full spikes suggested that at least some pacemaker cells are likely to be intrinsic oscillators. The relative amplitudes of the subthreshold and full spikes gave a lower bound to the gap junctional coupling coefficient of 0.01-0.08. Three drugs, called gap junction blockers for their mode of action in other preparations, caused immediate and substantial reduction in frequency, altered the phase lag between pairs of neurons, and later caused the spike amplitude to drop, without altering the spike timing precision. Thus we conclude that the high precision of the normal Pn rhythm does not require maximal gap junction conductances between neurons that have ordinary cellular precision. Rather, the spiking precision can be explained as an intrinsic cellular property while the gap junctions act to frequency- and phase-lock the network oscillations. PMID- 10669510 TI - Gap junction effects on precision and frequency of a model pacemaker network. AB - We investigated the precision of spike timing in a model of gap junction-coupled oscillatory neurons. The model incorporated the known physiology, morphology, and connectivity of the weakly electric fish's high-frequency and extremely precise pacemaker nucleus (Pn). Two neuron classes, pacemaker and relay cells, were each modeled with two compartments containing Hodgkin-Huxley sodium and potassium currents. Isolated pacemaker cells fired periodically, due to a constant current injection; relay cells were silent but slightly depolarized at rest. When coupled by gap junctions to other neurons, a model neuron, like its biological correlate, spiked at frequencies and amplitudes that were largely independent of current injections. The phase distribution in the network was labile to intracellular current injections and to gap junction conductance changes. The model predicts a biologically plausible gap junction conductance of 4-5 nS (200-250 MOmega). This results in a coupling coefficient of approximately 0.02, as observed in vitro. Network parameters were varied to test which could improve the temporal precision of oscillations. Increased gap junction conductances and larger numbers of cells (holding total junctional conductance per cell constant) both substantially reduced the coefficient of variation (CV = standard deviation/mean) of relay cell spike times by 74-85% and more, and did so with lower gap junction conductance when cells were contacted axonically compared with somatically. Pacemaker cell CV was only reduced when the probability of contact was increased, and then only moderately: a fivefold increase in the probability of contact reduced CV by 35%. We conclude that gap junctions facilitate synchronization, can reduce CV, are most effective between axons, and that pacemaker cells must have low intrinsic CV to account for the low CV of cells in the biological network. PMID- 10669511 TI - Modifications of seizure susceptibility in Drosophila. AB - In a given population, certain individuals are much more likely to have seizures than others. This increase in seizure susceptibility can lead to spontaneous seizures, such as seen in idiopathic epilepsy, or to symptomatic seizures that occur after insults to the nervous system. Despite the frequency of these seizure disorders in the human population, the genetic and physiological basis for these defects remains unclear. The present study makes use of Drosophila as a potentially powerful model for understanding seizure susceptibility in humans. In addition to the genetic and molecular advantages of using Drosophila, it has been found that seizures in Drosophila share much in common with seizures seen in humans. However, the most powerful aspect of this model lies in the ability to accurately measure seizure susceptibility across genotypes and over time. In the current study seizure susceptibility was quantified in a variety of mutant and wild-type strains, and it was found that genetic mutations can modulate susceptibility over an extremely wide range. This genetic modulation of seizure susceptibility apparently occurs without affecting the threshold of individual neurons. Seizure susceptibility also varied depending on the experience of the fly, decreasing immediately after a seizure and then gradually increasing over time. A novel phenomenon was also identified in which seizures are suppressed after certain high-intensity stimuli. These results demonstrate the utility of Drosophila as a model system for studying human seizure disorders and provide insights into the possible mechanisms by which seizure susceptibility is modified. PMID- 10669512 TI - Altered regulation of potassium and calcium channels by GABA(B) and adenosine receptors in hippocampal neurons from mice lacking Galpha(o). AB - To examine the role of G(o) in modulation of ion channels by neurotransmitter receptors, we characterized modulation of ionic currents in hippocampal CA3 neurons from mice lacking both isoforms of Galpha(o). In CA3 neurons from Galpha(o)(-/-) mice, 2-chloro-adenosine and the GABA(B)-receptor agonist baclofen activated inwardly rectifying K(+) currents and inhibited voltage-dependent Ca(2+) currents just as effectively as in Galpha(o)(+/+) littermates. However, the kinetics of transmitter action were dramatically altered in Galpha(o)(-/-) mice in that recovery on washout of agonist was much slower. For example, recovery from 2-chloro-adenosine inhibition of calcium current was more than fourfold slower in neurons from Galpha(o)(-/-) mice [time constant of 12.0 +/- 0.8 (SE) s] than in neurons from Galpha(o)(+/+) mice (time constant of 2.6 +/- 0.2 s). Recovery from baclofen effects was affected similarly. In neurons from control mice, effects of both baclofen and 2-chloro-adenosine on Ca(2+) currents and K(+) currents were abolished by brief exposure to external N-ethyl-maleimide (NEM). In neurons lacking Galpha(o), some inhibition of Ca(2+) currents by baclofen remained after NEM treatment, whereas baclofen activation of K(+) currents and both effects of 2-chloro-adenosine were abolished. These results show that modulation of Ca(2+) and K(+) currents by G protein-coupled receptors in hippocampal neurons does not have an absolute requirement for Galpha(o). However, modulation is changed in the absence of Galpha(o) in having much slower recovery kinetics. A likely possibility is that the very abundant Galpha(o) is normally used but, when absent, can readily be replaced by G proteins with different properties. PMID- 10669513 TI - Dynamic properties of recurrent inhibition in primary visual cortex: contrast and orientation dependence of contextual effects. AB - A fundamental feature of neural circuitry in the primary visual cortex (V1) is the existence of recurrent excitatory connections between spiny neurons, recurrent inhibitory connections between smooth neurons, and local connections between excitatory and inhibitory neurons. We modeled the dynamic behavior of intermixed excitatory and inhibitory populations of cells in V1 that receive input from the classical receptive field (the receptive field center) through feedforward thalamocortical afferents, as well as input from outside the classical receptive field (the receptive field surround) via long-range intracortical connections. A counterintuitive result is that the response of oriented cells can be facilitated beyond optimal levels when the surround stimulus is cross-oriented with respect to the center and suppressed when the surround stimulus is iso-oriented. This effect is primarily due to changes in recurrent inhibition within a local circuit. Cross-oriented surround stimulation leads to a reduction of presynaptic inhibition and a supraoptimal response, whereas iso-oriented surround stimulation has the opposite effect. This mechanism is used to explain the orientation and contrast dependence of contextual interactions in primary visual cortex: responses to a center stimulus can be both strongly suppressed and supraoptimally facilitated as a function of surround orientation, and these effects diminish as stimulus contrast decreases. PMID- 10669514 TI - Regional differences in hypoxic depolarization and swelling in hippocampal slices. AB - Pyramidal neurons in the CA1 region of the hippocampus are highly vulnerable to damage from hypoxia-ischemia, whereas neurons in the CA3 region and the dentate gyrus are more resistant. A similar pattern of vulnerability to loss of synaptic and membrane function occurs in the in vitro hippocampal slice preparation, suggesting that intrinsic factors are important in acute neuronal damage. Simultaneous recordings of DC potential and imaging of changes in light transmittance were made in slices from the middle one-third of the hippocampus to characterize the initiation and spread of depolarization and swelling during hypoxia-aglycemia. Hypoxic depolarization (HD) and associated optical changes were initiated simultaneously in the stratum oriens of the CA1 region and thereafter spread to the stratum radiatum of CA1 and later to the upper (inner) blade of the dentate gyrus. A decrease in light transmittance was associated consistently with depolarization in all regions (n = 22 slices). Investigation of the sequence of activation in intact slices showed that activation of the dentate gyrus arose independently of activation of the CA1 region. This was confirmed by recordings made from minislices in which CA1, CA3, and dentate regions were physically separated. HD and optical changes were never observed in the CA3 region, despite exposure to 40-60 min of combined hypoxia and aglycemia. In contrast, exposure to hypoxia after pretreatment of slices with altered tonicity or ion composition, which triggered episodes of spreading depolarization (SD), provoked depolarization and optical changes simultaneously in both CA1 and CA3 regions. Similarly, pretreatment with agents that cause severe metabolic impairment, such as dinitrophenol (DNP), also rendered the CA3 region vulnerable to subsequent hypoxia. This suggests that the CA3 region in hippocampal slices is normally resistant to HD and only becomes vulnerable after severe impairment of metabolic capacity. The similar order of vulnerability of in vitro and in vivo hippocampus to hypoxia-aglycemia supports the use of the hippocampal slice preparation to investigate early changes potentially contributing to hypoxic ischemic injury. PMID- 10669515 TI - Spatial distribution and characteristics of voltage-gated calcium signals within visual interneurons. AB - Most of our knowledge about insect calcium currents is derived from studies on cultured or dissociated somata. So far, only little data on calcium currents are available for neurons including their dendritic and presynaptic structures. Here we combined the switched-electrode voltage-clamp technique with optical recording using calcium-sensitive dyes in identified fly visual interneurons in vivo to characterize the voltage dependence and dynamics of calcium currents quantitatively and in a spatially resolved way. For all three cell types considered, i.e., centrifugal horizontal (CH), horizontal system (HS), and vertical system (VS) cells, the activation curve is rather flat and covers a voltage range from -60 to -20 mV in dendritic as well as presynaptic areas of the cells. The calcium increase is fastest for CH cells with a time constant of approximately 70 ms. In HS and VS cells, the time constant amounts to 400-700 ms. The calcium dynamics as determined in different regions of the cells are similar except for a small segment between the axon and the dendrite in HS and VS cells, where the calcium increase is significantly faster. In summary, the results show the existence of a low-voltage-activated calcium current with little or no inactivation in dendritic as well as presynaptic regions of fly lobula plate tangential cells. PMID- 10669516 TI - Mapping of IP(3)-mediated Ca(2+) signals in single human neuroblastoma SH-SY5Y cells: cell volume shaping the Ca(2+) signal. AB - Fast confocal laser-scanning microscopy was used to study spatiotemporal properties of IP(3)-mediated Ca(2+) release signals in human SH-SY5Y neuroblastoma cells. [Ca(2+)](i) increases were not affected by ryanodine (30 microgM) or caffeine (10 mM) and largely insensitive to removal of external Ca(2+), indicating predominance of IP(3)-induced Ca(2+) release. Ca(2+) signals evoked by high concentration (10 microM) of the muscarinic agonist carbachol appeared as self-propagating waves initiating in cell processes. At low carbachol concentrations (500 nM) Ca(2+) changes in most cells displayed striking spatiotemporal heterogeneity. The Ca(2+) response in the cell body was delayed and had a smaller amplitude and a slower rise time than that in processes. Ca(2+) changes in processes either occurred in a homogeneous manner throughout the whole process or were sometimes confined to hot spots. Regional differences in surface to-volume ratio appear to be critical clues that determine the spatiotemporal pattern of intracellular Ca(2+) release signals. PMID- 10669517 TI - Lateralized tinnitus studied with functional magnetic resonance imaging: abnormal inferior colliculus activation. AB - Tinnitus, the perception of sound in the absence of external stimuli, is a common and often disturbing symptom that is not understood physiologically. This paper presents an approach for using functional magnetic resonance imaging (fMRI) to investigate the physiology of tinnitus and demonstrates that the approach is effective in revealing tinnitus-related abnormalities in brain function. Our approach as applied here included 1) using a masking noise stimulus to change tinnitus loudness and examining the inferior colliculus (IC) for corresponding changes in activity, 2) separately considering subpopulations with particular tinnitus characteristics, in this case tinnitus lateralized to one ear, 3) controlling for intersubject differences in hearing loss by considering only subjects with normal or near-normal audiograms, and 4) tailoring the experimental design to the characteristics of the tinnitus subpopulation under study. For lateralized tinnitus subjects, we hypothesized that sound-evoked activation would be abnormally asymmetric because of the asymmetry of the tinnitus percept. This was tested using two reference groups for comparison: nontinnitus subjects and nonlateralized tinnitus subjects. Binaural noise produced abnormally asymmetric IC activation in every lateralized tinnitus subject (n = 4). In reference subjects (n = 9), activation (i.e., percent change in image signal) in the right versus left IC did not differ significantly. Compared with reference subjects, lateralized tinnitus subjects showed abnormally low percent signal change in the IC contralateral, but not ipsilateral, to the tinnitus percept. Consequently, activation asymmetry (i.e., the ratio of percent signal change in the IC ipsilateral versus contralateral to the tinnitus percept) was significantly greater in lateralized tinnitus subjects as compared with reference subjects. Monaural noise also produced abnormally asymmetric IC activation in lateralized tinnitus subjects. Two possible models are presented to explain why IC activation was abnormally low contralateral to the tinnitus percept in lateralized tinnitus subjects. Both assume that the percept is associated with abnormally high ("tinnitus-related") neural activity in the contralateral IC. Additionally, they assume that either 1) additional activity evoked by sound was limited by saturation or 2) sound stimulation reduced the level of tinnitus-related activity as it reduced the loudness of (i.e., masked) the tinnitus percept. In summary, this work demonstrates that fMRI can provide objective measures of lateralized tinnitus and tinnitus-related activation can be interpreted at a neural level. PMID- 10669519 TI - Human cerebellum plays an important role in memory-timed finger movement: an fMRI study. AB - The purpose of this study was to determine, by using functional magnetic resonance imaging, the areas of the brain activated during a memory-timed finger movement task and compare these with those activated during a visually cued movement task. Because it is likely that subjects engage in subvocalization associated with chronometric counting to achieve accurate timing during memory timed movements, the authors sought to determine the areas of the brain activated during a silent articulation task in which the subjects were instructed to reproduce the same timing as for the memory-timed movement task without any lip movements or vocalization. The memory-timed finger movement task induced activation of the anterior lobe of the cerebellum (lobules IV and V) bilaterally, the contralateral primary motor area, the supplementary motor area (SMA), the premotor area (PMA), the prefrontal cortex, and the posterior parietal cortex bilaterally, compared with the resting condition. The same areas in the SMA and left prefrontal cortex were activated during the silent articulation task compared with the resting condition. The anterior lobe of the cerebellum on both sides was also activated during the silent articulation task compared with the resting condition, but these activations did not reach statistical significance (P < 0.05 corrected). In addition, the anterior cerebellum on both sides showed significant activation during the memory-timed movement task when compared with the visually cued finger movement task. The visually cued finger movement task specifically activated the ipsilateral PMA and the intraparietal cortex bilaterally. The results indicate that the anterior lobe of the cerebellum of both sides, the SMA, and the left prefrontal cortex were probably involved in the generation of accurate timing, functioning as a clock within the CNS, and that the dorsal visual pathway may be involved in the generation of visually cued movements. PMID- 10669520 TI - Characteristics of plateau activity during the latent period prior to epileptiform discharges in slices from rat piriform cortex. AB - The deep piriform region has an unusually high seizure susceptibility. Voltage imaging previously located the sites of epileptiform discharge onset in slices of rat piriform cortex and revealed the spatiotemporal pattern of development of two types of electrical activity during the latent period prior to discharge onset. A ramplike depolarization (onset activity) appears at the site of discharge onset. Onset activity is preceded by a sustained low-amplitude depolarization (plateau activity) at another site, which shows little if any overlap with the site of onset. Because synaptic blockade at either of these two sites blocks discharges, it was proposed that both forms of latent period activity are necessary for the generation of epileptiform discharges and that the onset and plateau sites work together in the amplification of electrical activity. The capacity for amplification was examined here by studying subthreshold responses in slices of piriform cortex using two different in vitro models of epilepsy. Under some conditions electrically evoked responses showed a nonlinear dependence on stimulus current, suggesting amplification by strong polysynaptic excitatory responses. The sites of plateau and onset activity were mapped for different in vitro models of epilepsy and different sites of stimulation. These experiments showed that the site of plateau activity expanded into deep layers of neighboring neocortex in parallel with expansions of the onset site into neocortex. These results provide further evidence that interactions between the sites of onset and plateau activity play an important role in the initiation of epileptiform discharges. The site of plateau activity showed little variation with different stimulation sites in the piriform cortex, but when stimulation was applied in the endopiriform nucleus (in the sites of onset of plateau activity), plateau activity had a lower amplitude and became distributed over a much wider area. These results indicate that in the initiation of epileptiform discharges, the location of the circuit that generates plateau activity is not rigidly defined but can exhibit flexibility. PMID- 10669518 TI - GABA(B) receptors couple to potassium and calcium channels on identified lateral perforant pathway projection neurons. AB - Activation of presynaptic GABA(B) receptors inhibits neurotransmitter release at most cortical synapses, at least in part because of inhibition of voltage-gated calcium channels. One synapse where this is not the case is the lateral perforant pathway synapse onto dentate granule cells in the hippocampus. The current study was conducted to determine whether the neurons that make these synapses express GABA(B) receptors that can couple to ion channels. Perforant pathway projection neurons were labeled by injecting retrograde tracer into the dorsal hippocampus. The GABA(B) receptor agonist baclofen (10 microM) activated inwardly rectifying potassium channels and inhibited currents mediated by voltage-gated calcium channels in retrogradely labeled neurons in layer II of the lateral entorhinal cortex. These effects were reversed by coapplication of the selective GABA(B) receptor antagonist CGP 55845A (1 microM). Equivalent effects were produced by 100 microM adenosine, which inhibits neurotransmitter release at lateral perforant pathway synapses. The effects of baclofen and adenosine on inward currents were largely occlusive. These results suggest that the absence of GABA(B) receptor-mediated presynaptic inhibition at lateral perforant pathway synapses is not simply due to a failure to express these receptors and imply that GABA(B) receptors can either be selectively localized or regulated at terminal versus somatodendritic domains. PMID- 10669521 TI - Saccade-related activity in the parietal reach region. AB - In previous experiments, we showed that cells in the parietal reach region (PRR) in monkey posterior parietal cortex code intended reaching movements in an eye centered frame of reference. These cells are more active when an arm compared with an eye movement is being planned. Despite this clear preference for arm movements, we now report that PRR neurons also fire around the time of a saccade. Of 206 cells tested, 29% had perisaccadic activity in a delayed-saccade task. Two findings indicate that saccade-related activity does not reflect saccade planning or execution. First, activity is often peri- or postsaccadic but seldom presaccadic. Second, cells with saccade-related activity were no more likely to show strong saccadic delay period activity than cells without saccade-related activity. These findings indicate that PRR cells do not take part in saccade planning. Instead, the saccade-related activity in PRR may reflect cross-coupling between reach and saccade pathways that may be used to facilitate eye-hand coordination. Alternatively, saccade-related activity may reflect eye position information that could be used to maintain an eye-centered representation of intended reach targets across eye movements. PMID- 10669522 TI - Difference between visually and electrically evoked gaze saccades disclosed by altering the head moment of inertia. AB - Differences between gaze shifts evoked by collicular electrical stimulation and those triggered by the presentation of a visual stimulus were studied in head free cats by increasing the head moment of inertia. This maneuver modified the dynamics of these two types of gaze shifts by slowing down head movements. Such an increase in the head moment of inertia did not affect the metrics of visually evoked gaze saccades because their duration was precisely adjusted to compensate for these changes in movement dynamics. In contrast, the duration of electrically evoked gaze shifts remained constant irrespective of the head moment of inertia, and therefore their amplitude was significantly reduced. These results suggest that visually and electrically evoked gaze saccades are controlled by different mechanisms. Whereas the accuracy of visually evoked saccades is likely to be assured by on-line feedback information, the absence of duration adjustment in electrically evoked gaze shifts suggests that feedback information necessary to maintain their metrics is not accessible or is corrupted during collicular stimulation. This is of great importance when these two types of movements are compared to infer the role of the superior colliculus in the control of orienting gaze shifts. PMID- 10669523 TI - Modulation of dendritic action currents decreases the reliability of population spikes. AB - During synchronous action potential (AP) firing of CA1 pyramidal cells, a population spike (PS) is recorded in the extracellular space, the amplitude of which is considered a reliable quantitative index of the population output. Because the AP can be actively conducted and differentially modulated along the soma and dendrites, the extracellular part of the dendritic inward currents variably contributes to the somatic PS by spreading in the volume conductor to adjacent strata. This contribution has been studied by current-source density analysis and intracellular recordings in vivo during repetitive backpropagation that induces their selective fading. Both the PS and the ensemble action currents declined during high-frequency activation, although at different rates and timings. The decline was much stronger in dendrites than in the somatic region. At specific frequencies and for a short number of impulses the decrease of the somatic PS was neither due to fewer firing cells nor to decreased somatic action currents but to the blockade of dendritic action currents. The dendritic contribution to the peak of the somatic antidromic PS was estimated at approximately 30-40% and up to 100% at later times in the positive-going limb. The blockade of AP dendritic invasion was in part due to a decreased transfer of current from the soma that underwent a cumulative increase of conductance and slow depolarization during the train that eventually extended into the axon. The possibility of differential modulation of soma and dendritic action currents during APs should be checked when using the PS as a quantitative parameter. PMID- 10669524 TI - Early Life Influences and Interventions in Asthma. Introduction. PMID- 10669525 TI - Prevalence and etiology of asthma. AB - An increased understanding of the causes of asthma is coming from the international comparisons of asthma prevalence, particularly those from the European Community Respiratory Health Survey of asthma prevalence in adults and the International Study of Asthma and Allergies in Childhood. From these and other studies of asthma prevalence, it is possible to draw some tentative conclusions as to the patterns of asthma prevalence worldwide. There are five striking patterns: first, asthma prevalence is increasing worldwide; second, asthma is generally more common in Western countries and less common in developing countries; third, asthma is more prevalent in English-speaking countries; fourth, asthma prevalence is increasing in developing countries as they become more Westernized or communities become urbanized; and fifth, the prevalence of other allergic disorders may also be increasing worldwide. These five key features of the international patterns of asthma prevalence raise major questions about the role of "established" risk factors for the development of asthma. As a result, recent research has expanded to include the study of novel factors that may "program" the initial susceptibility to sensitization or contribute to the development of asthma independent of atopic sensitization. These include various exposures in utero, which are reflected in various perinatal factors measured at birth, and exposures (or lack of exposures) in the early years of life that may make the infant more susceptible to the subsequent development of asthma. These issues are now the focus of an intensive research effort worldwide, and the next few years are likely to see exciting advances in our understanding of the causes of asthma. PMID- 10669526 TI - Issues in understanding childhood asthma. AB - Asthma and related allergic disorders in childhood have increased considerably in prevalence over the last few decades. During the same period of increasing morbidity from childhood asthma in the community, there have been dramatic advances in understanding of the basic immunopathologic features of the disease and consequently the development of a far more rational approach to its treatment. The immunopathologic condition of eosinophil-mediated airway inflammation is established very early in the evolution of asthma in childhood. It may even antedate the onset of symptoms. The present state of the art dictates that early intervention with potent therapies cannot be justified on the basis of symptoms alone and may in any case have no influence on the natural history of the condition. This means that current cautious therapeutic guidelines should continue to be followed. However, with the development of more accurate markers predicting ongoing disease, it will be possible to evaluate a whole range of early interventions in the future. Much evidence, though indirect, points to the possibility that the only true prophylaxis that will affect the natural history of asthma will need to be commenced before clinical features are manifest. PMID- 10669527 TI - Mapping susceptibility genes for asthma and allergy. AB - Allergy and asthma are related conditions caused by a complex interaction of genetic factors and environmental influences. With family data from several different populations, linkage analysis has been performed and used to identify regions of the genome that contain susceptibility genes for these conditions. To date, 4 genome screens have been completed and have successfully identified several chromosomal locations that are likely to contain asthma and allergy genes. Many of these regions contain potential biologic candidate genes that modulate immunologic responses or airways inflammation. By focusing on the common regions that have been replicated in these 4 genome screens, the major susceptibility genes for asthma and allergy should be identified. This will lead to an improved understanding of pathogenic factors that lead to development or progression of asthma and allergic diseases. PMID- 10669528 TI - Strategies for analyzing genotype-phenotype relationships in asthma. AB - Asthma is a genetically complex disease with a multifaceted phenotype. Different approaches including population-based and family-based methods for evaluating genotype-phenotype relationships in asthma are discussed as well as the problems that may obscure these determinations. Examples of similar efforts in cystic fibrosis and breast cancer are considered in addition to interaction between causative genes and etiologically relevant environmental exposure. PMID- 10669529 TI - The pharmacogenetics of beta2-adrenergic receptors: relevance to asthma. AB - The beta(2)-adrenergic receptor (beta(2)AR) is the molecular target for beta agonists used in the treatment of asthma. In the human population, 4 polymorphisms of the beta(2)AR coding block have been found, 3 of which result in receptors that have different properties compared with wild-type. To date, clinical studies suggest that these beta(2)AR polymorphisms may alter asthmatic phenotype and the response to beta-agonist therapy, making these variants the first of undoubtedly several genetic loci that will ultimately be found that will provide for individualized therapy in asthma. PMID- 10669530 TI - Prenatal origins of allergic disease. AB - The prevalence of asthma and related allergic disorders has increased considerably over the last 25 years. Because genetic stock has not changed, environmental factors must have influenced the phenotype. Infants who experience the development of allergy already have an altered immune response at birth. We have investigated the development of immune responses during gestation and the effect of maternal allergen exposure during pregnancy and infant exposure in the first month of life on the development of allergy and disease. There was higher specific peripheral blood mononuclear cell proliferation to house dust mite and birch pollen in the third trimester compared with the second trimester, with the first positive responses seen at 22 weeks gestation. Maternal exposure to birch pollen after 22 weeks resulted in higher infant peripheral blood mononuclear cell responses to birch pollen at birth. Infants born at term, with at least 1 atopic parent with asthma, who experienced the development of allergic symptoms and positive skin prick test by 1 year of age had raised proliferative responses to house dust mite at birth compared with those infants with no symptoms. In genetically predisposed individuals, antenatal factors including maternal and thereby fetal exposure to allergens and materno-placental-fetal immunologic interactions are active in determining whether an allergic predisposition is manifested as disease. PMID- 10669531 TI - Viral and bacterial infections in the development and progression of asthma. AB - Viral respiratory infections produce wheezing illnesses in patients of all ages. In infancy, infections with respiratory syncytial virus and parainfluenza virus are the major cause of bronchiolitis and croup, whereas infections with common cold viruses such as rhinoviruses are the principal triggers for wheezing in older children and adults with asthma. In addition to causing increased wheezing in asthma, there is mounting evidence that infections early in childhood can affect the development of the immune system and thereby modify the risk for the subsequent development of allergies and asthma. Both of these effects appear to be mediated by virus-induced immune responses. Early during the course of viral infection, resident cells in the airway are activated in an antigen-independent fashion, triggering antiviral responses but also activating and recruiting cells to the airway that could contribute to airway obstruction and respiratory symptoms. Virus-specific T- and B-cell responses may also have dual effects in the presence of preexisting airway inflammation. Finally, there is evidence of synergistic interactions between allergen- and virus-induced airway inflammation. It is likely that greater definition of mechanisms of virus-induced inflammation will provide therapeutic targets for the treatment and possibly the prevention of allergies and asthma. PMID- 10669532 TI - The relevance of allergen exposure to the development of asthma in childhood. AB - Sensitization to 1 or more of the common indoor allergens has been consistently associated with asthma among children and young adults (odds ratios for asthma, 3 18). For dust mite and cockroach allergens, there is a dose response relationship between domestic exposure and sensitization. Given that allergen provocation can induce many of the features of asthma, the findings strongly suggest that there is a causal relationship between allergen exposure in the home and asthma. However, it remains unclear at what time the critical exposure occurs (ie, in infancy or later) and what role allergen exposure has played in the increasing prevalence and severity of asthma. Objective evidence of an immune response to allergens is generally not present until after 2 years of age. Viral infections play several different roles in asthma in childhood. In infancy, respiratory syncytial virus infection can induce bronchiolitis and set up recurrent wheezing over the next few years. However, the risk factors for this are maternal smoking and small lungs at birth, rather than allergy. By contrast, the role of rhinovirus in precipitating attacks in children and young adults is strongly associated with allergy. Thus the likely scenario is that allergen exposure over the first few years of life induces sensitization (ie, T(H2) cells and IgE antibodies). Continuing exposure can maintain inflammation in the nose and lungs. However, many other factors contribute to wheezing such that there is no simple relationship between allergen exposure and asthma. Nonetheless, it is clear that the changes that have increased asthma have acted on allergic children. PMID- 10669533 TI - Histopathologic features of early and progressive asthma. AB - During the last decade, morphologic studies on bronchial biopsy specimens have led to our present understanding of asthma as an inflammatory airways disease. However, little knowledge exists about the sequence of cellular events during the disease or of possible mucosal changes early in asthma. So far the primary cause, the site of damage, and the mechanisms inducing the inflammatory reaction remain to be elucidated. A multifactorial genetic susceptibility may be important for the development of asthma. Suggested factors that may trigger changes in the cells' morphologic and functional phenotype are viral infections, allergen exposure, maternal factors, diet, and smoking. Current evidence has implied that interactions between epithelial cells and the subepithelial connective tissue in the mucosa are important for normal homeostatic balance. Changes in airway epithelial phenotype possibly resulting from altered gene expression in its lining cells may be very important even as a first line change in asthma. PMID- 10669534 TI - Structural consequences of airway inflammation in asthma. AB - Asthma represents a chronic inflammatory process of the airways followed by healing, the end-result of which is an altered structure referred to as a remodeling of the airways. Repair usually involves 2 distinct processes: regeneration (which is the replacement of injured tissue by parenchymal cells of the same type) and replacement by connective tissue and its eventual maturation into scar tissue. In many instances both processes contribute to the healing response and inflammation. In asthma the processes of cell dedifferentiation, migration, differentiation, and maturation and connective tissue deposition can be followed either by complete or altered restitution of airway structure and function, the latter often seen as fibrosis and increase in smooth muscle and mucus gland mass. These features result in an increased resistance to airflow, particularly when there is bronchial contraction and bronchial hyperresponsiveness. The effect on airflow is compounded by the presence of increased mucous secretion and inflammatory exudate, which not only blocks the airway passages but also causes an increased surface tension that favors airway closure. PMID- 10669535 TI - The role of proteoglycans in the regulation of airways inflammation and airways remodelling. PMID- 10669536 TI - Asthma: A disease of inflammation and repair. AB - Mucosal inflammation and more recently airway remodeling continue to be a focus of interest when considering both the pathophysiology and treatment of asthma. Although a number of candidate genes relevant to inflammatory cell action have been identified and linked to atopy and airway hyperresponsiveness, it is important to understand genetic factors that might determine the extent of tissue remodeling. The mechanisms regulating the allergic responses in the airways are complex, involving antigen presenting cells and T lymphocytes, which process antigens and orchestrate the response, and mast cells and eosinophils as effector cells. Abundant evidence also points to a proinflammatory role for structural cells, including epithelial and endothelial cells, and smooth muscle. Because of the complex nature of the inflammatory changes in asthma, the exact relation between individual inflammatory cells and their mediators on the one hand and hyperresponsiveness of the airways and clinical manifestations on the other remains unclear. The same applies to the phenomenon of airway remodeling because it is uncertain exactly how the different components of restructuring affect the airway physiology. If progress is to be made in the treatment of asthma, further efforts will be needed to understand the regulation and link between the mechanisms causing inflammation and those leading to fibrotic changes. PMID- 10669537 TI - Mast-cell responses in the development of asthma. AB - Many cells participate in the pathogenesis of asthmatic inflammation. The mast cell is localized at the interface of the internal and external environment within the lung where it may respond to allergens and other exogenous stimuli. The activation of mast cells leads to the release of mediators that contribute to the early phase of asthmatic inflammation. Mast-cell-derived products may also contribute to the late-phase asthmatic response. This review summarizes the developmental biologic features of the mast cell, its receptor-mediated activation, and its range of preformed, newly synthesized, and induced mediators that contribute to asthmatic inflammation. PMID- 10669538 TI - Natural history of chronic asthma and its long-term effects on pulmonary function. AB - Although asthma is a disease that has intrigued physicians since antiquity, its natural history has been incompletely determined. It has long been held that the presence of asthma, per se, does not carry with it any long-term deterioration in lung function, but recently this view has been challenged, and it has become fashionable to define asthma as being only partially reversible. At present, there are limited data to support such a view. All of the available information indicates that the vast majority of patients do not experience the development of a progressive decline in pulmonary mechanics or appear to be at risk for a diminution in life expectancy. PMID- 10669539 TI - Interaction between the growing lung and asthma: role of early intervention. AB - Asthma is a syndrome where an imbalance exists between the forces that maintain airway patency and the forces that act to narrow, or close, airways. The child with asthma is a particular problem because of the rapid growth of the lung during growth that leaves it vulnerable. There is some evidence that asthma leads to impaired lung function in children because those children with untreated asthma show a loss of lung growth velocity. For unclear reasons, asthma is more frequent in boys. What drugs to use to treat childhood asthma is uncertain. Data show that glucocorticoids prevent the structure of the lung from fully developing. In children, the rationale for early intervention seems clear, but the exact means and criteria for initiation of the intervention are uncertain. Finally, childhood asthma raises fundamental issues and questions that are unique to the child with asthma and presents unique and many unresolved treatment dilemmas. PMID- 10669540 TI - Molecular mechanisms of IgE regulation. AB - IgE antibody plays an important role in allergic diseases. IgE synthesis by B cells requires two signals. The first signal is delivered by the cytokines IL-4 or IL-13, which target the Cepsilon gene for switch recombination. The second signal is delivered by interaction of the B cell surface antigen CD40 with its ligand (CD40L) expressed on activated T cells. This activates deletional switch recombination. We review the molecular mechanisms of IL-4 and CD40 signaling that lead to IgE isotype switching and discuss the implications for intervening to abort or suppress the IgE antibody response. PMID- 10669541 TI - The consideration of immunotherapy in the treatment of allergic asthma. AB - Immunotherapy has undergone rigorous trials to assess its therapeutic benefit in the treatment of allergic respiratory disease. The tools of molecular biology have provided a framework with which to begin to understand the mechanistic effects of immunotherapy on the underlying inflammatory component of allergic respiratory disease. The clinical relevance of these observations belies our understanding of allergic inflammation as the subsoil for the development of abnormal airway physiology, heightened bronchial reactivity, and the development of chronic asthmatic symptoms. Immunotherapy provides the potential to downregulate this inflammatory cascade, reduce IgE antibody production, and attenuate symptoms. Conceptually, early intervention of allergic disease holds the most promise as a therapeutic intervention capable of arresting the progression of the disease, altering the severity of the disease, and/or preventing the development of the respiratory disease process. PMID- 10669542 TI - The effects of cromolyn sodium and nedocromil sodium in early asthma prevention. AB - The possibility of irreversible obstruction and therefore the need for early intervention is being much debated. Some investigators suggested that delay in starting inhaled corticosteroids will result in irreversible obstruction. Our own long-term study, specifically designed to detect irreversible obstruction showed that a step-wise approach (starting with cromolyn sodium and switching to inhaled corticosteroids if clinical control and pulmonary function tests are not satisfactorily controlled) resulted in an increase in pulmonary function and not a deterioration. There was no evidence that a delay in starting inhaled corticosteroids will result in irreversible obstruction or clinical worsening. However, delay in starting cromolyn sodium in patients treated with bronchodilators alone did result in worsening pulmonary function tests and worse clinical outcomes. One study from Finland and another study from Australia came to the same conclusion. Even though some studies with cromolyn sodium did not show benefit in the first year of life, other studies did show a good response. The choice between nonsteroidal drugs, such as cromolyn sodium and inhaled corticosteroids as first-line drugs, has to be made on the risk/benefit ratio of these drugs. Although in severe asthma inhaled corticosteroids have greater efficacy, in mild-to-moderate asthma there is comparable efficacy, and the nonsteroidal drugs have better safety. A step-wise approach is still a logical approach PMID- 10669543 TI - Early interventions in asthma with inhaled corticosteroids. AB - We have earlier shown epithelial damage in the airway mucosa in patients with asthma. Later other structural changes have been recognized in asthma, such as deposition of collagen and tenascin in the subepithelial basement membrane and changes in the laminin subchain composition. These processes are modified by an inflammatory process in the airways. Both the United States National Institutes of Health and the British Thoracic Society guidelines on the management of asthma emphasize the need for early use of anti-inflammatory drugs. Many clinical studies that used airway biopsy specimens have shown a decrease in airway inflammatory cell numbers after inhaled corticosteroid therapy. However, there is very little information on the effects of asthma medication on the structural components of the airways. Both the synthesis and degradation of many extracellular matrix components may be affected by the disease process and the drugs resulting in altered remodeling and gene expression in the airways. Because there are only a few studies that try to identify early changes in asthma, it is not known whether the anti-inflammatory treatment of asthma proposed by the guidelines is started early enough. PMID- 10669544 TI - New immunopharmacologic approaches to asthma: role of cytokine antagonism. AB - This article involves an elucidation of the potential inflammatory mechanisms associated with the treatment of allergic disease and asthma, and the possibility of cytokine antagonism as a potential therapeutic mechanism for the treatment of those diseases. There is a review of the role of cytokines in the allergic process and a description of a number of studies done with the capacity of certain cytokine antagonists to develop potential amelioration of immune dysregulation in asthma and atopic states. PMID- 10669545 TI - Signal transduction and cellular radiation responses. AB - Exposure of cells to ionizing radiation results in complex cellular responses resulting in cell death and altered proliferation states. The underlying cytotoxic, cytoprotective and cellular stress responses to radiation are mediated by existing signaling pathways, activation of which may be amplified by intrinsic cellular radical production systems. These signaling responses include the activation of plasma membrane receptors, the stimulation of cytoplasmic protein kinases, transcriptional activation, and altered cell cycle regulation. From the data presented, there is increasing evidence for the functional links between cellular signal transduction responses and DNA damage recognition and repair, cell survival, or cell death through apoptosis or reproductive mechanisms. PMID- 10669546 TI - Alpha-particle-induced changes in the stability and size of DNA. AB - The effect of alpha-particle radiation on the thermal stability and size of calf thymus DNA molecules in deoxygenated aqueous solutions was investigated by thermal transition spectrophotometry, pulsed-field gel electrophoresis, and standard agarose gel electrophoresis. The thermal transition of DNA from helix to coil was studied through analysis of the UV A(260) absorbance. The results obtained for alpha particles of mean LET of 128 keV microm(-1) reveal a dual dose response: a tendency for thermal stability of the DNA helix at "low" doses, followed by an increasing instability at higher doses. The same phenomenon was observed for the mean molecular weight of DNA molecules exposed to alpha particles. The results reported here for alpha particles in the low-dose region of 0-16 Gy are consistent with our previous hypothesis of inter- and intramolecular interactions of a covalent character in gamma-irradiated DNA molecules in the dose region of 0-4 Gy. PMID- 10669547 TI - Iodine-125 decay in a synthetic oligodeoxynucleotide. I. Fragment size distribution and evaluation of breakage probability. AB - Lobachevsky, P. N. and Martin, R. F. Iodine-125 Decay in a Synthetic Oligodeoxynucleotide. I. Fragment Size Distribution and Evaluation of Breakage Probability. Incorporation of (125)I-dC into a defined location of a double stranded oligodeoxynucleotide was used to investigate DNA breaks arising from decay of the Auger electron-emitting isotope. Samples of the oligodeoxynucleotide were also labeled with (32)P at either the 5' or 3' end of either the (125)I-dC containing (so-called top) or opposite (bottom) strand and incubated in 20 mM phosphate buffer or the same buffer plus 2 M dimethylsulfoxide at 4 degrees C during 18-20 days. The (32)P-end-labeled fragments produced by (125)I decays were separated on denaturing polyacrylamide gels, and the (32)P activity in each fragment was determined by scintillation counting after elution of fragments from the gel. The relative fragment size distributions were then normalized on a per decay basis and converted to a distribution of single-strand break probabilities as a function of distance from the (125)I-dC. The results of three to five experiments for each of eight possible combinations of labels and incubation conditions are presented as a table showing the relative numbers of (32)P counts in different fragments as well as graphs of normalized fragment size distributions and probabilities of breakage. The average numbers of single-strand breaks per (125)I decay are 3. 3 and 3.7 in the top strand and 1.3 and 1.5 in the bottom strand with and without dimethylsulfoxide, respectively. Every (125)I decay event produces a break in the top strand, and breakage of the bottom strand occurs in 75-80% of the events. Thus a double-strand break is produced by (125)I decay with a probability of approximately 0.8. PMID- 10669548 TI - Iodine-125 decay in a synthetic oligodeoxynucleotide. II. The role of auger electron irradiation compared to charge neutralization in DNA breakage. AB - Lobachevsky, P. N. and Martin, R. F. Iodine-125 Decay in a Synthetic Oligodeoxynucleotide. II. The Role of Auger Electron Irradiation Compared to Charge Neutralization in DNA Breakage. The dramatic chemical and biological effects of the decay of DNA-incorporated (125)I stem from two consequences of the Auger electron cascades associated with the decay of the isotope: high local deposition of radiation energy from short-range Auger electrons, and neutralization of the multiply charged tellurium atom. We have analyzed the extensive data reported in the companion paper (Radiat. Res. 153, 000-000, 2000), in which DNA breakage was measured after (125)I decay in a 41-bp oligoDNA. The experimental data collected under scavenging conditions (2 M dimethylsulfoxide) were deconvoluted into two components denoted as radiation and nonradiation, the former being attributed to energy deposition by Auger electrons. The contribution of the components was estimated by adopting various assumptions, the principal one being that DNA breakage due to the radiation mechanism is dependent on the distance between the decaying (125)I atom and the cleaved deoxyribosyl unit, while the nonradiation mechanism, associated with neutralization of the multiply charged tellurium atom, contributes equally at corresponding nucleotides starting from the (125)I-incorporating nucleotide. Comparison of the experimental data sets collected under scavenging and nonscavenging (without dimethylsulfoxide) conditions was used to estimate the radiation-scavengeable component. Our analysis showed that the nonradiation component plays the major role in causing breakage within 4-5 nucleotides from the site of (125)I incorporation and produces about 50% of all single-stranded breaks. This overall result is consistent with the relative amounts of energy associated with Auger electrons and the charged tellurium atom. However, the nonradiation component accounts for almost four times more breaks in the top strand, to which the (125)I is bound covalently, than in the bottom strand, thus suggesting an important role of covalent bonds in the energy transfer from the charged tellurium atom. The radiation component dominates at the distances beyond 8-9 nucleotides, and 36% of the radiation-induced breaks are scavengeable. PMID- 10669549 TI - Inducible repair and intrinsic radiosensitivity: a complex but predictable relationship? AB - Two groups have proposed a simple linear relationship between inducible radioresistance in a variety of mammalian cells and their intrinsic radiosensitivity at 2 Gy (Lambin et al., Int.J. Radiat. Biol. 69, 279-290, 1996; Alsbeih and Raaphorst, unpublished results, 1997). The inducible repair response (IRR) is quantified as a ratio, alpha(S)/alpha(R), i.e. the slope in the hypersensitive low-dose region, alpha(S), relative to the alpha(R) term of the classical linear-quadratic formula. These proposals imply that the intrinsic radiosensitivity at clinically relevant doses is directly linked to the cell's ability to mount an adaptive response as a result of exposure to very low doses of radiation. We have re-examined this correlation and found that the more extensive data set now available in the literature does not support the contention of a simple linear relationship. The two parameters are correlated, but by a much more complex relationship. A more logical fit is obtained with a log-linear equation. A series of log-linear curves are needed to describe the correlation between IRR and SF2, because of the spectrum of alpha/beta ratios among the cell lines and hence the confounding effect of the beta term at a dose of 2 Gy. The degree of repair competence before irradiation starts could also be a major factor in the apparent magnitude of the amount of repair induced. There appears to be a systematic difference in the data sets from different series of cell lines that have been obtained using flow cytometry techniques in the laboratory in Vancouver and using dynamic microscope imaging at the Gray Laboratory. We suggest that the use of a brief exposure to a laser beam in flow cytometry before the cells are irradiated might itself partially induce a stress response and change the DNA repair capacity of the cells. The clinical consequences of the relationship for predicting the benefits of altered fractionation schedules are discussed. [ru5] PMID- 10669550 TI - Radiation dose dependences in the atomic bomb survivor cancer mortality data: a model-free visualization. AB - Chomentowski, M., Kellerer, A. M. and Pierce, D. A. Radiation Dose Dependences in the Atomic Bomb Survivor Cancer Mortality Data: A Model-Free Visualization. The standard approach to obtaining nominal risk coefficients for radiation-related cancer involves fitting linear or linear-quadratic dose-response functions. This is usually complemented by a more direct visualization where the data are subdivided into distinct dose categories and the effect level is quantified for each of these categories. Such model-free computations, however, can be quite dependent on the arbitrary choice of the cutpoints in dose. The method proposed here largely avoids this arbitrariness by choosing a dose category width-constant on a log scale-to obtain the desired degree of smoothing, and then superimposing results for all placements of the resulting log-dose grids. The method is applied to leukemia and solid cancer mortality of the A-bomb survivors. PMID- 10669551 TI - Apoptosis and cell cycle progression in an acidic environment after irradiation. AB - Apoptosis and cell cycle progression in HL60 cells irradiated in an acidic environment were investigated. Apoptosis was determined by TUNEL staining, PARP cleavage, DNA fragmentation, and flow cytometry. The majority of the apoptosis that occurred in HL60 cells after 4 Gy irradiation took place after G(2)/M-phase arrest. When irradiated with 12 Gy, a fraction of the cells underwent apoptosis in G(1) and S phases while the rest of the cells underwent apoptosis in G(2)/M phase. The apoptosis caused by 4 and 12 Gy irradiation was transiently suppressed in medium at pH 7.1 or lower. An acidic environment was found to perturb progression of irradiated cells through the cell cycle, including progression through G(2)/ M phase. Thus it was concluded that the suppression of apoptosis in the cells after 4-12 Gy irradiation in acidic medium was due at least in part to a delay in cell cycle progression, particularly the prolongation of G(2)/M-phase arrest. Irradiation with 20 Gy indiscriminately caused apoptosis in all cell cycle phases, i.e. G(1), S and G(2)/M phases, rapidly in neutral pH medium and relatively slowly in acidic pH medium. The delay in apoptosis in acidic medium after 20 Gy irradiation appeared to result from mechanisms other than prolonged G(2)/ M-phase arrest. PMID- 10669552 TI - Dose-dependent biphasic accumulation of TP53 protein in normal human embryo cells after X irradiation. AB - The effects of various doses of X radiation on the kinetics of accumulation of TP53 protein (formerly known as p53) were examined in normal human embryo cells. We found that the rate of accumulation of TP53 protein was biphasic at X-ray doses between 1 and 4 Gy, while monophasic accumulation was observed after X irradiation with doses higher than 6 Gy. The first phase of accumulation was detected within 1 h after irradiation, and a second phase of accumulation was detected between 6 and 12 h after irradiation. The induction of CDKN1A (formerly known as p21(WAF1/CIP1)) and MDM2 proteins was also biphasic after doses of 4 Gy or less, while monophasic accumulation was observed after 6 Gy or higher. We found that the proteasome inhibitor ALLN increased the constitutive level of TP53 protein, and no change was observed in the TP53 level after X irradiation when cells were treated with ALLN. These results indicate that the dose-dependent accumulation of TP53 is due to an inhibition of TP53 degradation, and that the induction of MDM2 might be responsible in part for the different kinetics of accumulation of TP53. PMID- 10669553 TI - Dose-dependent changes in the spectrum of mutations induced by ionizing radiation. AB - We examined the influence of dose on the spectrum of mutations induced at the hypoxanthine guanine phosphoribosyltransferase (Hprt) locus in Chinese hamster ovary (CHO) cells. Independent CHO-K1 cell mutants at the Hprt locus were isolated from cells exposed to 0, 0.5, 1.5, 3.0 and 6.0 Gy (137)Cs gamma rays, and the genetic changes responsible for the mutations were determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. We observed dose-dependent changes in mutation spectra. At low doses, the principal radiation-induced mutations were point mutations. With increasing dose, multibase deletion mutations became the predominant mutation type such that by 6.0 Gy, there were almost three times more deletion mutations than point mutations. The dose response for induction of point mutations was linear while that for multibase deletions fit a linear-quadratic response. There was a biphasic distribution of deletion sizes, and different dose responses for small compared to large deletions. The frequency of large (>36 kb) total gene deletions increased exponentially, implying that they develop from the interaction between two independent events. In contrast, the dose response for deletion mutations of less than 10 kb was nearly linear, suggesting that these types of mutations develop mostly from single events and not the interactions between two independently produced lesions. The observation of dose-dependent changes in radiation-induced mutation spectra suggests that the types of alterations and therefore the risks from low-dose radiation exposure cannot be easily extrapolated from high-dose effects. PMID- 10669554 TI - Inducible heat-shock protein 70 is involved in the radioadaptive response. AB - Park, S-H., Lee, S-J., Chung, H-Y., Kim, T-H., Cho, C-K., Yoo, S-Y. and Lee, Y-S. Inducible Heat-Shock Protein 70 Is Involved in the Radioadaptive Response. The thermoresistant (TR) clone of radiation-induced fibrosarcoma (RIF) cells showed an adaptive response, i.e. a reduced effect, after exposure to a higher challenging dose (4 Gy) when the priming dose (1 cGy) was given 4 or 7 h earlier, but RIF cells did not. Since inducible Hsp70 expression was different in cells of these two cell lines, the role of inducible Hsp70 in the adaptive response was examined. When inducible Hsp70 was transfected into RIF cells, the adaptive response was acquired. Transfection of inducible Hsp70 to NIH 3T3 mouse embryo cells also conferred radioresistance to the cells as assayed by clonogenic survival, [(3)H]thymidine incorporation, and an ELISA cell death detection kit. An increased tendency for the induction of an adaptive response was also observed. Interestingly, basal levels of Ca(2+)-dependent and independent Pkc activities were increased by transfection with inducible Hsp70 compared to those of control vector cells. Irradiation with gamma rays induced activation of Pkc within minutes in control vector cells, while transfection with inducible Hsp70 did not. Cellular redistribution to particulate fractions of Pkca, d and z after exposure gamma rays also was not detected. Furthermore, radioresistance by transfection with inducible Hsp70, as tested by clonogenic survival, disappeared after pretreatment with Pkc inhibitors, 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H7), prolonged treatment with 12-O-tetradecanoylphorbol-13 acetate (TPA), and GF109203X. Taken together, these data suggest that radioresistance inducible by Hsp70 is associated with an elevated level of Pkc activity. PMID- 10669555 TI - Effect of a combination of mild-temperature hyperthermia and nicotinamide on the radiation response of experimental tumors. AB - Ogawa, A., Griffin, R. J. and Song, C. W. Effect of a Combination of Mild Temperature Hyperthermia and Nicotinamide on the Radiation Response of Experimental Tumors. The effect of mild-temperature hyperthermia and nicotinamide individually or combined on tumor radiosensitivity was investigated with SCK tumors grown s.c. in the right hind limbs of A/J mice. An i.p. injection of nicotinamide at 50-250 mg/kg slightly enhanced the cell killing caused by 10-20 Gy of ionizing radiation as determined by the in vivo/in vitro tumor excision assay. Treatment of tumors with mild-temperature hyperthermia at 41.5 degrees C for 60 min prior to tumor irradiation was significantly more effective than nicotinamide and the combination of nicotinamide and hyperthermia was far more effective than nicotinamide or hyperthermia alone in enhancing radiation-induced cell killing. Radiation-induced tumor growth delay was enhanced by a factor of 1.2 by 50 mg/kg nicotinamide, 2.1 by hyperthermia, and 3.6 by the combination of nicotinamide and hyperthermia. Taking these results and those of our previous studies together, we conclude that mild-temperature hyperthermia increases tumor blood flow and oxygenation and that combining mild-temperature hyperthermia and nicotinamide is more effective than either of these alone in increasing tumor radiosensitivity. PMID- 10669556 TI - Immune effects of low-dose radiation: short-term induction of thymocyte apoptosis and long-term augmentation of T-cell-dependent immune responses. AB - We and others have shown that low-dose X or gamma irradiation of mice leads to an increase in their survival after a subsequent lethal high-dose irradiation. The greatest increase in radioresistance appears at a fixed window of dose and time, e.g. 8 weeks after 5-10 cGy or 2 weeks after 50 cGy preirradiation. We show that low-dose irradiation induces thymocyte apoptosis with a maximal level at 6 h postirradiation that returns to background levels after 24 h. At the same time, we observed no morphological alteration of splenocytes and no early modification of the intensity of T-cell-dependent immune responses as measured by plaque forming cell (PFC) counts. Nevertheless, we found that PFCs were increased 2 weeks after 50 cGy irradiation, which is the same time at which mice expressed the optimal increase in survival after a second lethal irradiation. We also examined thymocyte apoptosis and spleen PFCs in mice subjected to other stress inducing pretreatments. Our results emphasize the existence of a lag time between the time of low-dose irradiation in vivo and the appearance of radioresistance. A mechanism that interconnects an environmental stimulus with the response of the animal is proposed based on the evidence presented here and reported in the literature. PMID- 10669557 TI - Early radiation effects on muscarinic receptor-induced secretory responsiveness of the parotid gland in the freely moving rat. AB - Although the salivary glands have a low rate of cell turnover, they are relatively radiosensitive. To study the possible mechanism behind this inherent radiosensitivity, a rat model was developed in which saliva can be collected after local irradiation of the parotid gland without the use of anesthetics or stressful handling. Saliva secretion was induced by the partial muscarinic receptor agonist pilocarpine (0.03-3 mg/kg) with or without pretreatment with the beta-adrenoceptor antagonist propranolol (2.5 mg/kg), or the full muscarinic receptor agonist methacholine (0.16-16 mg/min), and measured during 5 min per drug dose before and 1, 3, 6 and 10 days after irradiation. The maximal secretory response induced by pilocarpine plus propranolol was increased compared to that with pilocarpine alone but did not reach the level of methacholine-induced secretion, which was about five times higher. One day after irradiation a decrease in maximal pilocarpine-induced secretion was observed (-22%) using the same dose of pilocarpine that induces 50% of the maximal response (ED(50)), in both the absence and presence of propranolol, indicating that the receptor-drug interaction was not affected by the radiation at this time. The secretory response to methacholine 1 day after irradiation, however, was normal. At day 3 after irradiation, the maximal methacholine-induced secretion was also affected, whereas pilocarpine (+/-propranolol)-induced maximal secretion decreased further. At day 6 after irradiation, maximal secretory responses had declined to approximately 50% regardless of the agonist used, whereas ED(50) values were still unaffected. No net acinar cell loss was observed within the first 10 days after irradiation, and this therefore could not account for the loss in function. The results indicate that radiation does not affect cell number or receptor-drug interaction, but rather signal transduction, which eventually leads to the impaired response. We hypothesize that the early radiation effect, within 3 days, may be membrane damage affecting the receptor-G-protein signaltransfer. Later critical damage, however, is probably of a different nature and may be located in the second-messenger signal transduction pathway downstream from the G protein, not necessarily involving cellular membranes. PMID- 10669558 TI - Cellular commitment to radiation-induced apoptosis. AB - The basic elements of the machinery of programmed cell death (apoptosis) are built into all mammalian cells and are conserved evolutionarily from nematodes to humans. The workshop on Commitment to Radiation-Induced Apoptosis at the 11th International Congress of Radiation Research in Dublin, Ireland reviewed recent information regarding the basic molecular mechanisms which are fundamental to the understanding of the process of apoptosis after treatment with ionizing radiation and some other agents. PMID- 10669559 TI - Protease inhibitors: current status and future prospects. PMID- 10669560 TI - A novel, potent, and selective 5-HT(7) antagonist: (R)-3-(2-(2-(4-methylpiperidin 1-yl)ethyl)pyrrolidine-1-sulfonyl) phen ol (SB-269970). PMID- 10669561 TI - alpha-Substituted N-(sulfonamido)alkyl-beta-aminotetralins: potent and selective neuropeptide Y Y5 receptor antagonists. PMID- 10669562 TI - Discovery of a novel dopamine transporter inhibitor, 4-hydroxy-1-methyl-4-(4 methylphenyl)-3-piperidyl 4-methylphenyl ketone, as a potential cocaine antagonist through 3D-database pharmacophore searching. Molecular modeling, structure-activity relationships, and behavioral pharmacological studies. AB - A novel, fairly potent dopamine transporter (DAT) inhibitor, 4-hydroxy-1-methyl-4 (4-methylphenyl)-3-piperidyl 4-methylphenyl ketone (3, K(i) values of 492 and 360 nM in binding affinity and inhibition of dopamine reuptake, respectively), with significant functional antagonism against cocaine and a different in vitro pharmacological profile from cocaine at the three transporter sites (dopamine, serotonin, and norepinephrine) was discovered through 3D-database pharmacophore searching. Through structure-activity relationships and molecular modeling studies, we found that hydrophobicity and conformational preference are two additional important parameters that determine affinity at the DAT site. Chemical modifications of the lead compound (3) led to a high affinity analogue (6, K(i) values of 11 and 55 nM in binding affinity and inhibition of dopamine reuptake, respectively). In behavioral pharmacological testing, 6 mimics partially the effect of cocaine in increasing locomotor activity in mice but lacks cocaine-like discriminative stimulus effect in rats. Taken together, these data suggest that 6 represents a promising lead for further evaluations as potential therapy for the treatment of cocaine abuse. PMID- 10669563 TI - Structural basis of the thrombin selectivity of a ligand that contains the constrained arginine mimic (2S)-2-amino-(3S)-3-(1-carbamimidoyl- piperidin-3-yl) propanoic acid at P1. AB - We have studied the thrombin and trypsin complexed structures of a pair of peptidomimetic thrombin inhibitors, containing different P1 fragments. The first has arginine as its P1 fragment, and the second contains the constrained arginine mimic (2S)-2-amino-(3S)-3-(1-carbamimidoyl-piperidin-3-yl)-propano ic acid (SAPA), a fragment known to enhance thrombin/trypsin selectivity of inhibitors. On the basis of an analysis of the nonbonded interactions present in the structures of the trypsin and thrombin complexes of the two inhibitors, the calculated accessible surfaces of the enzymes and inhibitors in the four complexes, data on known structures of trypsin complexes of inhibitors, and factor Xa inhibitory potency of these compounds, we conclude that the ability of this arginine mimic to increase thrombin selectivity of an inhibitor is mediated by its differential interaction with the residue at position 192 (chymotrypsinogen numbering). Thrombin has a glutamic acid at residue 192, and trypsin has a glutamine. The analysis also suggests that this constrained arginine mimic, when present in an inhibitor, might enhance selectivity against other trypsin-like enzymes that have a glutamine at residue position 192. PMID- 10669564 TI - Design and synthesis of piperazine-based matrix metalloproteinase inhibitors. AB - A new generation of cyclic matrix metalloproteinase (MMP) inhibitors derived from dl-piperazinecarboxylic acid has been described. The design involves: incorporation of hydroxamic acid as the bidentate chelating agent for catalytic Zn(2+), placement of a sulfonamide group at the 1N-position of the piperazine ring to fill the S1' pocket of the enzyme, and finally attachment of diverse functional groups at the 4N-position to optimize potency and peroral absorption. A unique combination of all three elements produced inhibitor 20 with high affinity for MMPs 1, 3, 9, and 13 (24, 18, 1.9, and 1.3 nM, respectively). X-ray crystallography data obtained for MMP-3 cocrystallized with 20 gave detailed information on key binding interactions defining an overall scaffold geometry for piperazine-based MMP inhibitors. PMID- 10669565 TI - Molecular docking reveals a novel binding site model for fentanyl at the mu opioid receptor. AB - The ligand binding modes of a series of fentanyl derivatives are examined using a combination of conformational analysis and molecular docking to the mu-opioid receptor. Condensed-phase molecular dynamics simulations are applied to evaluate potential relationships between ligand conformation and fentanyl substitution and to generate probable "bioactive" structures for the ligand series. Automated docking of the largely populated solution conformers identified a common binding site orientation that places the N-phenethyl group of fentanyl deep in a crevice between transmembrane (TM) helices II and III while the N-phenylpropanamide group projected toward a pocket formed by TM-III, -VI, and -VII domains. An analysis of the binding modes indicates the most potent fentanyl derivatives adopt an extended conformation both in solution and in the bound state, suggesting binding affinity may depend on the conformational preferences of the ligands. The results are consistent with ligand binding data derived from chimeric and mutant receptor studies as well as structure-activity relationship data reported on a wide range of fentanyl analogues. The binding site model is also compared to that of N phenethylnormorphine. An overlay of the bound conformation of the opiate and cis 3-methylfentanyl shows the N-phenethyl groups occupy equivalent binding domains in the receptor. While the cationic amines of both ligand classes were found docked to an established anchor site (D149 in TM-III), no overlap was observed between the N-phenylpropanamide group and the remaining components of the opiate scaffold. The unique binding mode(s) proposed for the fentanyl series may, in part, explain the difficulties encountered in defining models of recognition at the mu-receptor and suggest opioid receptors may display multiple binding epitopes. Furthermore, the results provide new insight to the design of experiments aimed at understanding the structural basis to the differential selectivities of ligands at the mu-, delta-, and kappa-opioid receptors. PMID- 10669566 TI - Derivation of pharmacophore and CoMFA models for leukotriene D(4) receptor antagonists of the quinolinyl(bridged)aryl series. AB - The present work focuses on the study of the three-dimensional (3D) structural requirements for the leukotriene D(4) (LTD(4)) antagonistic activity of compounds having the basic quinolinyl(bridged)aryl framework. An approach combining pharmacophore mapping, molecule alignment, and CoMFA models was used to derive a hypothesis for a series of LTD(4) antagonists having the basic diaryl-bridged framework. In this compound series, the produced pharmacophore hypotheses have shown to yield molecule alignments suitable to derive valuable CoMFA models. Model selection focused on (1) obtention of coherent modeling results, (2) consistency with the available SAR data, and (3) ability to predict the activity of an independent set of congeneric molecules. This approach resulted in a combined pharmacophore and CoMFA model that can generally represent the antagonistic activity within a log unit of the measured value for compounds of the series. The resulting pharmacophore (model C) consists of an acidic or negative ionizable function (AC), a hydrogen-bond acceptor (HBA), and three hydrophobic regions (HY) and produces chemically meaningful alignments with the most active compounds of the series mapping the pharmacophore in a extended energetically favorable conformation. PMID- 10669567 TI - Successful virtual screening of a chemical database for farnesyltransferase inhibitor leads. AB - Virtual screening of chemical databases is an emerging approach in drug discovery that uses computers to dock chemicals into the active site of a drug target to identify leads through evaluation of binding affinities of the chemicals. However, there are concerns about the validity and scope of the reported virtual screens due to lack of studies to show that randomly selected chemicals are not equally active and due to the fact that metalloproteins were rarely used as drug targets. We have performed a virtual screening of a chemical database to identify prototypic inhibitors of farnesyltransferase (FT) with zinc present in the active site. Among the 21 compounds identified by computers, four inhibited FT in vitro with IC(50) values in the range from 25 to 100 microM. The most potent inhibitor also inhibited FT in human lung cancer cells. In contrast, none of 21 randomly selected compounds have an IC(50) lower than 100 microM. The results demonstrate the validity of virtual screening and the feasibility of applications of this approach to metalloprotein drug targets, such as matrix metalloproteinases, farnesyltransferase, and HIV-1 integrase, for the treatments of cardiovascular diseases, cancers, and AIDS. PMID- 10669568 TI - Syntheses and structure-activity relationships of 5,6,7, 8-tetrahydro-5,5,8,8 tetramethyl-2-quinoxaline derivatives with retinoic acid receptor alpha agonistic activity. AB - In the course of our studies on retinoic acid receptor (RAR) agonists, we have designed and synthesized a series of quinoxaline derivatives. One of them, 4-[5 (5,6,7,8-tetrahydro-5,5,8, 8-tetramethyl-2-quinoxalinyl)-1H-2-pyrrolyl]benzoic acid (3a), which possesses a 2,5-disubstituted pyrrole moiety, showed selectivity for the RARalpha receptor and exerted highly potent cell-differentiating activity on HL-60 cells. PMID- 10669569 TI - Synthesis, molecular modeling, and pharmacological testing of bis-quinolinium cyclophanes: potent, non-peptidic blockers of the apamin-sensitive Ca(2+) activated K(+) channel. AB - The synthesis and pharmacological testing of two series of novel bis-quinolinium cyclophanes as blockers of the apamin-sensitive Ca(2+)-activated K(+) (SK(Ca)) channel are presented. In these cyclophanes the two 4-aminoquinolinium groups are joined at the ring N atoms (linker L) and at the exocyclic N atoms (linker A). In those cases where A and L contain two or more aromatic rings each, the activity of the compound is not critically dependent upon the nature of the linkers. When A and L each have only one benzene ring, the blocking potency changes dramatically with simple structural variations in the linkers. One of these smaller cyclophanes having A = benzene-1,4-diylbis(methylene) and L = benzene-1, 3-diylbis(methylene) (3j, 6,10-diaza-1,5(1,4)-diquinolina-3(1,3),8(1, 4) dibenzenacyclodecaphanedium tritrifluoroacetate, UCL 1684) has an IC(50) of 3 nM and is the most potent non-peptidic SK(Ca) channel blocker described to date. Conformational analysis on the smaller cyclophanes using molecular modeling techniques suggests that the differences in the blocking potencies of the compounds may be attributable to their different conformational preferences. PMID- 10669570 TI - Synthesis and in vitro and in vivo functional studies of ortho-substituted phenylpiperazine and N-substituted 4-N-(o-methoxyphenyl)aminopiperidine analogues of WAY100635. AB - WAY100635 (2), N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl)cyclohe xanecarboxamide, is a silent serotonin 5-HT(1A) antagonist, which is now widely used to study the 5-HT(1A) receptor both in vivo and in vitro. In this paper, we describe the synthesis and in vitro (5-HT(1A) affinity and pA(2) values at guinea pig ileum strips) and in vivo (hypothermia and ultrasonic vocalization) pharmacology at the serotonin 5-HT(1A) receptor of several closely related analogues of 2. Test compounds 12 and 14, in which the arylpiperazine moiety of 2 has been replaced by an arylaminopiperidine moiety, showed no affinity or antagonistic activity at the 5-HT(1A) receptor. Substitution of the o-methoxy group of 2 by larger fluoroalkoxy or sulfonyloxy substituents did not alter the in vitro or in vivo pharmacology to any great extent; in vivo both the fluoropropyl analogue 5 and the triflate analogue 7 are equipotent to WAY100635 itself. The O-desmethyl analogue 3 proved to be the most potent antagonist at the serotonin 5-HT(1A) postsynaptic receptor sites in this series. PMID- 10669571 TI - Water-soluble phosphate prodrugs of 1-propargyl-8-styrylxanthine derivatives, A(2A)-selective adenosine receptor antagonists. AB - Water-soluble prodrugs of potent, A(2A)-selective adenosine receptor (AR) antagonists were prepared. 8-(m-Bromostyryl)-3, 7-dimethyl-1-propargylxanthine (BS-DMPX, 11) and the analogous 8-(m-methoxystyryl)xanthine derivative (MS-DMPX, 5b) were used as starting points. It was found that polar functional groups suitable for the attachment of a prodrug moiety were tolerated on the styryl ring and even better on the 3-substituent. 8-(m-Hydroxystyryl)-DMPX (7) and 3-(3 hydroxypropyl)-8-(m-methoxystyryl)-1-propargylxanthine (5e, MSX-2) were the most potent and A(2A)-selective compounds and were selected for prodrug formation. For the preparation of 5e a new ring-closure method was applied. Treatment of 6-amino 1-(3-hydroxypropyl)-5-(m-methoxycinnamoylamino)-3-propa rgylur acil with hexamethyldisilazane at high temperature resulted in higher yields of the target xanthine than the standard ring-closure procedure using sodium hydroxide. Phosphate prodrugs were prepared by classical phosphorylation using phosphorus oxychloride and alternatively by using a phosphoramidite method. Phosphates of the aliphatic alcohol 5e could be obtained by both methods in similar yields. The phenolic compound 7, however, could be phosphorylated only by using the phosphoramidite method. The disodium salts of the phosphate prodrugs exhibited high water solubility (8-(m-methoxystyryl)-7-methyl-3-[3-O-phosphatylpropyl]-1- propargylxan thine disodium salt, 9b: 17 mM, 9 mg/mL). Prodrug 9b was found to be stable in aqueous solution (pH 7) but readily cleaved by phosphatases to liberate 5e (MSX-2). Compound 5e showed high affinity for rat A(2A) AR (K(i) = 8 nM), human recombinant A(2A) AR (K(i) = 5 nM), and human native A(2A) AR (K(i) = 15 nM) and was highly selective versus rat A(1) AR (110-fold), human recombinant A(2A) AR (500-fold), human A(2B) AR (>2000-fold), and human A(3) AR (>2000-fold). PMID- 10669572 TI - 4-(1,3-Dimethoxyprop-2-ylamino)-2,7-dimethyl-8-(2, 4-dichlorophenyl)pyrazolo[1,5 a]-1,3,5-triazine: a potent, orally bioavailable CRF(1) receptor antagonist. AB - Structure-activity studies in the pyrazolo[1,5-a]-1,3,5-triazine series led to the discovery that compound 11i (DMP696) is a potent hCRF(1) receptor antagonist (K(i) = 1.7 nM vs 7.5 nM for alpha-hel-CRF(9-41), hCRF(1) adenylate cyclase IC(50) = 82 nM vs 286 nM for alpha-hel-CRF(9-41)). Compound 11i has excellent oral pharmacokinetic profiles in rats and dogs (37% and 50% oral bioavailabilities, respectively). This compound displays good activity in the rat situational anxiety model (MED = 3 mg/kg (po)), whereas a literature standard 1 (CP154526-1) was inactive (MED > 30 mg/kg (po)). Analogue 11i reduced stereotypical mouth movements in rhesus monkeys by 50% at 21 mg/kg (po) using the human intruder paradigm. Overall, the profile of pyrazolotriazine 11i indicates that hCRF(1) receptor antagonists may be anxiolytic agents, which have reduced motor side effect profiles. PMID- 10669573 TI - Design of cancer-specific antitumor agents based on aziridinylcyclopent[b]indoloquinones. AB - The merits of N-unsubstituted indoles and cyclopent[b]indoles as DNA-directed reductive alkylating agents are described. These systems represent a departure from N-substituted and pyrrolo[1, 2-a]-fused systems such as the mitomycins and mitosenes. The cyclopent[b]indole-based aziridinylquinone system, when bearing an acetate leaving group with or without an N-acetyl group, was cytotoxic and displayed significant in vivo activity against syngeneic tumor implants. These analogues were superior to the others studied in terms of both high specificity for the activating enzyme DT-diaphorase and high percent DNA alkylation. Alkylation by a quinone methide intermediate as well as by the aziridinyl group could lead to cross-linking. The possible metabolites of the most active indole species were prepared and found to retain cytotoxicity, suggesting that in vivo activity could be sustained. The indole systems in the present study display selectivity for melanoma and, depending on the substituents present, selectivity for non-small-cell lung, colon, renal, and prostate cancers. The cancer specificities observed are believed to pertain to differential substrate specificities for DT-diaphorase. PMID- 10669574 TI - Acid-catalyzed degradation of clarithromycin and erythromycin B: a comparative study using NMR spectroscopy. AB - One of the major drawbacks in the use of the antibiotic erythromycin A is its extreme acid sensitivity, leading to degradation in the stomach following oral administration. The modern derivative clarithromycin degrades by a different mechanism and much more slowly. We have studied the pathway and kinetics of the acid-catalyzed degradation of clarithromycin and of erythromycin B, a biosynthetic precursor of erythromycin A which also has good antibacterial activity, using (1)H NMR spectroscopy. Both drugs degrade by loss of the cladinose sugar ring and with similar rates of reaction. These results suggest that erythromycin B has potential as an independent therapeutic entity, with superior acid stability compared with erythromycin A and with the advantage over clarithromycin of being a natural product. PMID- 10669575 TI - Drug delivery systems based on trimethyl lock lactonization: poly(ethylene glycol) prodrugs of amino-containing compounds. AB - A novel methodology for the synthesis of poly(ethylene glycol) (PEG) prodrugs of amino-containing compounds has been developed which is based on the trimethyl lock lactonization reaction. These PEG-modified double prodrugs are water soluble, and by selective modification of the specifier or trigger, plasma half lives can be adjusted at will to result in a wide range of values. Facile syntheses of ester, carbonate, and carbamate functionalities were accomplished and combined with greater or lesser degrees of steric hindrance in the spacer group, or on the aromatic framework, to achieve predictable ranges of drug concentration in plasma. In vivo screening of PEG prodrugs was done using a M109 syngeneic solid mouse tumor model. One of the PEG-daunorubicin prodrugs, with a half-life of 2 h, was evaluated in an in vivo solid tumor panel and found to be more efficacious against ovarian tumors (SKOV3) than equivalent amounts of daunorubicin. PMID- 10669576 TI - Potent and selective non-peptidic inhibitors of endothelin-converting enzyme-1 with sustained duration of action. AB - Potent and selective non-peptidic inhibitors of human endothelin-converting enzyme-1 (ECE-1) have been designed as potential modulators of endothelin (ET-1) production in vivo. Because of its unique structural characteristics and long duration of action in vivo, the dual ECE-1 and neutral endopeptidase 24.11 (NEP) inhibitor, CGS 26303, was selected as an attractive lead for further optimization of potency and selectivity. Replacement of the P(1)' biphenyl substituent of CGS 26303 by a conformationally restricted 3-dibenzofuranyl group led to more potent and more selective ECE-1 inhibitors, such as the tetrazole 27. The remarkable effect of this P(1)' modification allowed for the first time phosphonomethylcarboxylic acids, such as 29, to display both potent (IC(50) = 22 nM) and selective (104-fold vs NEP) ECE-1 inhibition. Chemoenzymatic syntheses of the new alpha-amino acid (S)-3-dibenzofuran-3-ylalanine intermediate were developed, and improved procedures to generate substituted alpha aminoalkylphosphonic acids were devised to support the production of various analogues. Although additional gains in intrinsic ECE-1 inhibitory potency could occasionally be achieved by addition of a P(1) side chain, these compounds (e.g. 43a) showed poor functional activity in vivo in the big ET-1 pressor test. Phosphonoalkyl dipeptides featuring 3-dibenzofuranyl groups in both the P(1)' and P(2)' positions were also very potent ECE-1 inhibitors, albeit lacking the desired selectivity against NEP. Functionally, 27and 29 were the two most efficacious compounds from this study, producing sustained inhibition of ECE-1 activity in rats, as measured by their ability to block the hypertensive effects induced by big ET-1. This profile was similar to that of a potent ET(A)/ET(B) dual receptor antagonist, SB 209670. Due to their favorable in vitro and in vivo profiles, 27 (CGS 34043) and 29 (CGS 35066) constitute new pharmacological tools useful in assessing the role of ECE-1 in pathological conditions. PMID- 10669577 TI - Antimalarial activity of compounds interfering with Plasmodium falciparum phospholipid metabolism: comparison between mono- and bisquaternary ammonium salts. AB - On the basis of a previous structure-activity relationship study, we identified some essential parameters, e.g. electronegativity and lipophilicity, required for polar head analogues to inhibit Plasmodium falciparum phospholipid metabolism, leading to parasite death. To improve the in vitro antimalarial activity, 36 cationic choline analogues consisting of mono-, bis-, and triquaternary ammonium salts with distinct substituents of increasing lipophilicity were synthesized. For monoquaternary ammonium salts, an increase in the lipophilicity around nitrogen was beneficial for antimalarial activity: IC(50) decreased by 1 order of magnitude from trimethyl to tripropyl substituents. Irrespective of the polar head substitution (methyl, ethyl, hydroxyethyl, pyrrolidinium), increasing the alkyl chain length from 6 to 12 methylene groups always led to increased activity. The highest activity was obtained for the N,N,N-tripropyl-N-dodecyl substitution of nitrogen (IC(50) 33 nM). Beyond 12 methylene groups, the antimalarial activities of the compounds decreased slightly. The structural requirements for bisquaternary ammonium salts in antimalarial activity were very similar to those of monoquaternary ammonium salts, i.e. polar head steric hindrance and lipophilicity around nitrogen (methyl, hydroxyethyl, ethyl, pyrrolidinium, etc.). In contrast, with bisquaternary ammonium salts, increasing the lipophilicity of the alkyl chain between the two nitrogen atoms (from 5 to 21 methylene groups) constantly and dramatically increased the activity. Most of these duplicated molecules had activity around 1 nM, and the most lipophilic compound synthesized exhibited an IC(50) as low as 3 pM (21 methylene groups). Globally, this oriented synthesis produced 28 compounds out of 36 with an IC(50) lower than 1 microM, and 9 of them had an IC(50) in the nanomolar range, with 1 compound in the picomolar range. This indicates that developing a pharmacological model for antimalarial compounds through choline analogues is a promising strategy. PMID- 10669578 TI - New selective and potent 5-HT(1B/1D) antagonists: chemistry and pharmacological evaluation of N-piperazinylphenyl biphenylcarboxamides and biphenylsulfonamides. AB - A series of new analogues of N-[4-methoxy-3-(4-methylpiperazin-1-yl)phenyl] 2' methyl-4'-(5-methyl-1,2,4-oxadiazol-3-yl)biphenyl-4-carboxamide (1; GR127935) as potent and selective 5-HT(1B/1D) antagonists were synthesized and evaluated pharmacologically. Their receptor binding profiles were comparable to that of 1. The 1,3,4-oxadiazole isomer 2 and the 4'-aminocarbonyl and 4'-amidinyl analogues (9 and 10) of 1 had higher affinities at the rat 5-HT(1B) receptor (IC(50) = 0.93, 1. 3, and 0.5 nM, respectively) and calf 5-HT(1D) receptor (IC(50) = 37, 10, and 3 nM, respectively) than did 1 (1.6 and 52 nM for rat 5-HT(1B) and calf 5 HT(1D) receptors, respectively). In the functional in vitro testing of 5 HT(1B/1D) antagonistic properties, 2, 9, 10, 11b (O-demethylated derivative of 2), 13a (O-methylsulfonyl analogue of 2), and 16 (which differs from 2 with a sulfonamide linker) showed more pronounced effects in the K(+)-induced 5-HT release in the cortex of guinea pig than did 1 and 3 (SB224289). Compounds 2, 9, and 10 were equally potent as 1 in rabbit saphenous vein model (pA(2) > 9). A biochemical study of 2 with in vivo microdialysis in the rat brain showed that it is capable of augmenting citalopram (a selective serotonin reuptake inhibitor, SSRI) induced 5-HT release in rat ventral hippocampus, while preventing the decrease in acetylcholine release elicited by citalopram administration. The molecular structure of 2 was determined by single-crystal X-ray analysis. The log P and log D values of these compounds were calculated. This study contributes to the SAR study of N-piperazinylphenyl biphenylcarboxamides as selective and potent 5-HT(1B/1D) antagonists. PMID- 10669579 TI - Integrin-like proteins in Candida spp. and other microorganisms. AB - The vertebrate integrins provide a paradigm for cell surface proteins involved in adhesion and morphogenesis. However, homologs of integrins have been found in more primitive organisms. This review will discuss the evidence for surface proteins in Candida albicans and Candida tropicalis that contain motifs reminiscent of integrins and will analyze the contributions of one of these proteins, Int1p, to adhesion, morphogenesis, and virulence. Other microorganisms thought to express integrin-like proteins will also be addressed. PMID- 10669580 TI - To perforate a leaf of grass. PMID- 10669581 TI - High levels of gene flow and heterozygote excess characterize Rhizoctonia solani AG-1 IA (Thanatephorus cucumeris) from Texas. AB - To date, much of the genetics of the basidiomycete Thanatephorus cucumeris (anamorph = Rhizoctonia solani) remains unknown. Here, we present a population genetics study using codominant markers to augment laboratory analyses. Seven single-copy nuclear RFLP markers were used to examine 182 isolates of Rhizoctonia solani AG-1 IA collected from six commercial rice fields in Texas. Thirty-six multilocus RFLP genotypes were identified. Population subdivision analyses indicated a high degree of gene flow/migration between the six geographic populations. Tests for Hardy-Weinberg equilibrium (HWE) among the 36 multilocus RFLP genotypes revealed that four of the seven loci did not significantly differ from HWE. Subsequent analysis demonstrated that departures from HWE at the three remaining loci were due to an excess of heterozygotes. Data presented here suggest that R. solani AG-1 IA is actively outbreeding (heterothallic). Possible explanations for heterozygote excess, which was observed at all seven RFLP loci, are discussed. PMID- 10669582 TI - Construction of a bacterial artificial chromosome library of Phytophthora infestans and transformation of clones into P. infestans. AB - A bacterial artificial chromosome (BAC) library of Phytophthora infestans was constructed in a derivative of pBELOBACII that had been modified by adding a npt selectable marker gene for transforming P. infestans. A total library of 8 genome equivalents was generated and 16,128 clones with inserts averaging 75 kb (4.9 genome equivalents) were individually picked and stored as an arrayed library in microtiter plates. This coverage was confirmed by screening the library for 11 DNA loci by colony hybridization and by polymerase chain reaction of DNA pools. Transformation of P. infestans with BAC clones containing inserts of 93 to 135 kb was demonstrated. The efficiency of transformation with most BACs was noticeably higher than that with smaller plasmids. Detailed analyses of transformants obtained with a 102-kb BAC indicated that entire inserts were present in about one-quarter of the transformants. PMID- 10669583 TI - Evidence that MRas1 and MRas3 proteins are associated with distinct cellular functions during growth and morphogenesis in the fungus Mucor racemosus. AB - The filamentous fungus Mucor racemosus provides a simple and unique model system for defining the function of individual ras genes in a gene family which is closely related to mammalian ras genes. The current study was designed to investigate the role of Mras1 and Mras3 in different stages of fungal morphogenesis, including sporangiospore germination, sporulation, and dimorphic transitions. The overall patterns of Mras1 and Mras3 transcript and protein accumulation were markedly different but, in general, transcripts and proteins were present at low levels during spherical growth and their accumulated level increased severalfold during polar growth (germ tube emergence and elongation). In contrast to Mras1, relatively high levels of Mras3 transcript accumulated during sporulation and MRas3 protein accumulated in sporangiospores. Transformation of M. racemosus with an activated allele of Mras3 reduced growth rate during aerobic sporangiospore germination, while a dominant-negative allele of Mras3 caused a 40% decrease in viable asexual spores. An activated allele of Mras1 increased growth rate during sporangiospore germination but neither activated nor dominant-negative alleles of Mras1 affected total number of asexual spores. Expression of MRas3 and MRas1 proteins appear to be subject to different regulatory mechanisms: exogenous dibutyryl-cAMP and fusidienol caused a strong repression of the level of MRas3 protein (but not MRas1) concurrent with the inhibition of polar growth. Differential posttranslational modification and intracellular localization of MRas1 and MRas3 proteins were also observed. The data strongly suggest that Mras3 and Mras1 play different roles in regulation of cell growth and morphogenesis in Mucor. PMID- 10669584 TI - Invasive hyphal growth in Wangiella dermatitidis is induced by stab inoculation and shows dependence upon melanin biosynthesis. AB - Stab inoculation of agar medium with yeasts of the human pathogen Wangiella dermatitidis resulted in induction of invasive hyphae. Mechanical penetration of agar was indicated by the observation that an increase in medium gel strength slowed the rate of substrate invasion. A melanized wild-type strain (8656) exhibited much faster invasive growth through 2-8% agar than three melanin deficient mutants. Inhibition of melanin synthesis in strain 8656 using tricyclazole resulted in a decrease in its rate of invasive growth, while scytalone restored melanin synthesis in the albino mel3 strain and boosted its rate of invasive growth. Earlier research established that cellular melanization is also associated with invasive hyphal growth in the mouse brain, and infections with strain 8656 are invariably lethal. Together, these in vitro and in vivo data indicate that biomechanical characteristics of fungi may be important determinants of virulence and disease progression in human and animal mycoses. PMID- 10669585 TI - Blue light signaling chains in Phycomyces: phototransduction of carotenogenesis and morphogenesis involves distinct protein kinase/phosphatase elements. AB - Carotenogenesis and morphogenesis represent two of the several responses sensitive to blue light which characterize the lower eukaryote Phycomyces blakesleeanus. Speculating that reversible phosphorylation may be an intracellular event beyond the photoperception step, we resorted to the use of first-choice inhibitors of protein phosphatases and protein kinases. The mycelial beta-carotene content of dark-grown cultures was induced by all agents administered, while the morphogenic output showed the typical trend effected by light only with one of the protein kinase inhibitors. Our data provide convincing evidence that protein phosphorylation plays a regulatory role in photocarotenogenesis and photomorphogenesis of Phycomyces. According to the model we propose, the putative signaling elements involved are anticipated to have a repressive function in the dark so that the responses are maintained in the "off" mode until the moment photon information has to flow through the regulatory circuit. PMID- 10669586 TI - Major surface glycoprotein genes from Pneumocystis carinii f. sp. ratti. AB - Pneumocystis carinii occurs in a variety of mammals, each of which harbors one or more genetically distinct "special forms" of the microbe. Laboratory rats can be infected by two special forms, P. carinii f. sp. ratti and P. carinii f. sp. carinii. P. carinii f. sp. carinii has a variable antigen, the major surface glycoprotein (MSG), the expression of which is controlled by genetic recombination. Recombination may involve the CRJE, a 23-bp DNA sequence element invariant among P. carinii f. sp. carinii MSG genes. To better understand the role of the CRJE in MSG gene expression and to explore the possible role of MSG in P. carinii infection in rats, P. carinii f. sp. ratti MSG genes were studied. These genes were found to be related to MSG genes of P. carinii f. sp. carinii, but less so than MSG genes from P. carinii f. sp. carinii are to each other. P. carinii f. sp. ratti MSG genes were present throughout the genome and were expressed as an abundant mRNA species slightly smaller than that found in P. carinii f. sp. carinii. P. carinii f. sp. ratti MSG transcripts included a CRJE like sequence only 78% identical to the CRJE of P. carinii f. sp. carinii. Comparison of MSG proteins from the two rat special forms of P. carinii to those from human, ferret, and mouse P. carinii did not support the hypothesis that growth in the rat lung requires certain primary MSG peptide sequences. PMID- 10669587 TI - The frost gene of Neurospora crassa is a homolog of yeast cdc1 and affects hyphal branching via manganese homeostasis. AB - The Neurospora crassa mutant frost has a hyperbranching phenotype that can be corrected by adding Ca(2+), suggesting that characterization of this gene might clarify the mechanism of Ca(2+)-dependent tip growth. The wild-type allele was cloned by sib selection using protoplasts from arthroconidia. RFLP analysis revealed that the cloned DNA fragment mapped to the fr locus. The nucleotide sequence of genomic and cDNA was determined. The deduced amino acid sequence showed homology to the Saccharomyces cerevisiae CDC1 protein, implicated in manganese homeostasis. The fr mutant was sensitive to Mn(2+), and a revertant allele whose product differs by one amino acid was tolerant to Mn(2+). Mn(2+) depletion induced the wild-type strain to hyperbranch, resulting in a morphology similar to that of fr. The fr mutant was also sensitive to calcineurin inhibitors. These results suggest that fr is involved in Mn(2+) homeostasis and point to a role for Mn(2+) in Neurospora branching. PMID- 10669589 TI - Cumulative organism index volumes 26-28 PMID- 10669588 TI - Phylogenetic analysis of a dataset of fungal 5.8S rDNA sequences shows that highly divergent copies of internal transcribed spacers reported from Scutellospora castanea are of ascomycete origin. AB - Using a dataset comprising 5.8S rDNA sequences from a wide range of fungi, we show that some sequences reported recently from the arbuscular mycorrhizal (AM) fungus Scutellospora castanea most likely originate from Ascomycetes. Other ITS and 5.8S sequences which were previously reported are confirmed as being clearly of mycorrhizal origin and are variable within one isolate of S. castanea. However, these results mean that previous conclusions which were drawn regarding the heterokaryotic status of AM fungal spores remain unproven. We provide an enlarged 5.8S rDNA dataset that can be used to check ITS sequences for conflicts with well-established phylogenies of the organisms that they were obtained from. PMID- 10669590 TI - The serine protease inhibitor canonical loop conformation: examples found in extracellular hydrolases, toxins, cytokines and viral proteins. AB - Methods for the prediction of protein function from structure are of growing importance in the age of structural genomics. Here, we focus on the problem of identifying sites of potential serine protease inhibitor interactions on the surface of proteins of known structure. Given that there is no sequence conservation within canonical loops from different inhibitor families, we first compare representative loops to all fragments of equal length among proteins of known structure by calculating main-chain RMS deviation. Fragments with RMS deviation below a certain threshold (hits) are removed if residues have solvent accessibilities appreciably lower than those observed in the search structure. These remaining hits are further filtered to remove those occurring largely within secondary structure elements. Likely functional significance is restricted further by considering only extracellular protein domains. By comparing different canonical loop structures to the protein structure database, we show that the method is able to detect previously known inhibitors. In addition, we discuss potentially new canonical loop structures found in secreted hydrolases, toxins, viral proteins, cytokines and other proteins. We discuss the possible functional significance of several of the examples found, and comment on implications for the prediction of function from protein 3D structure. PMID- 10669591 TI - Stabilization of poliovirus by capsid-binding antiviral drugs is due to entropic effects. AB - When poliovirus attaches to its receptor or is heated in hypotonic buffers, the virion undergoes an irreversible conformational transition from the native 160 S (or N) particle to the 135 S (or A) particle, which is believed to mediate cell entry. The first-order rate constants for the thermally induced transition have been measured as a function of temperature for virus alone and for complexes of the virus with capsid-binding drugs that inhibit the receptor and thermally mediated conversion. Although the drugs have minimum inhibitory concentrations (MIC) that differ by almost three orders of magnitude, the activation energies for the N to A transition for the drug complexes (145 kcal/mol) were indistinguishable from each other or from that of the virus alone. We conclude that the antiviral activity of these drugs derives from a novel mechanism in which drug-binding stabilizes the virions through entropic effects. PMID- 10669592 TI - Structure of human neutral endopeptidase (Neprilysin) complexed with phosphoramidon. AB - Neutral endopeptidase is a mammalian type II integral membrane zinc-containing endopeptidase, which degrades and inactivates a number of bioactive peptides. The range of substrates cleaved by neutral endopeptidase in vitro includes the enkephalins, substance P, endothelin, bradykinin and atrial natriuretic factor. Due to the physiological importance of neutral endopeptidase in the modulation of nociceptive and pressor responses there is considerable interest in inhibitors of this enzyme as novel analgesics and anti-hypertensive agents. Here we describe the crystal structure of the extracellular domain (residues 52-749) of human NEP complexed with the generic metalloproteinase inhibitor phosphoramidon at 2.1 A resolution. The structure reveals two multiply connected folding domains which embrace a large central cavity containing the active site. The inhibitor is bound to one side of this cavity and its binding mode provides a detailed understanding of the ligand-binding and specificity determinants. PMID- 10669593 TI - Solution structure and dynamics of the DNA-binding domain of the adipocyte transcription factor FREAC-11. AB - Transcription factors of the forkhead type share a highly conserved DNA-binding domain of about 100 amino acid residues. FREAC-11, expressed in adipocytes, belongs to this class. Here, we report on NMR studies that established the three dimensional structure of the FREAC-11, DNA-binding domain. Although apparent similarities to the structures of other members within the forkhead family are observed, the structure also reveals some remarkable differences. Along with the complementary dynamics, the data provide insight into the fundamentals of sequence specificity within a highly conserved motif. PMID- 10669594 TI - Salmonella enteritidis fimbriae displaying a heterologous epitope reveal a uniquely flexible structure and assembly mechanism. AB - Two distinct Salmonella fimbrins, AgfA and SefA, comprising thin aggregative fimbriae SEF17 and SEF14, respectively, were each genetically engineered to carry PT3, an alpha-helical 16-amino acid Leishmania T-cell epitope derived from the metalloprotease gp63. To identify regions within AgfA and SefA fimbrins amenable to replacement with this epitope, PCR-generated chimeric fimbrin genes were constructed and used to replace the native chromosomal agfA and sefA genes in Salmonella enteritidis. Immunoblot analysis using anti-SEF17 and anti-PT3 sera demonstrated that all ten AgfA chimeric fimbrin proteins were expressed by S. enteritidis under normal growth conditions. Immunoelectron microscopy confirmed that eight of the AgfA::PT3 proteins were effectively assembled into cell surface exposed fimbriae. The PT3 replacements in AgfA altered Congo red (CR) binding, cell-cell adhesion and cell surface properties of S. enteritidis to varying degrees. However, these chimeric fimbriae were still highly stable, being resistant to proteinase K digestion and requiring harsh formic acid treatment for depolymerization. In marked contrast to AgfA, none of the chimeric SefA proteins were expressed or assembled into fimbriae. Since each PT3 replacement constituted over 10% of the AgfA amino acid sequence and all ten replacements collectively represented greater than 75% of the entire AgfA primary sequence, the ability of AgfA to accept large sequence substitutions and still assemble into fibers is unique among fimbriae and other structural proteins. This structural flexibility may be related to the novel fivefold repeating sequence of AgfA and its recently proposed structure Proper formation of chimeric fimbrial fibers suggests an unusual assembly mechanism for thin aggregative fimbriae which tolerates aberrant structures. This study opens a range of possibilities for Salmonella thin aggregative fimbriae as a carrier of heterologous epitopes and as an experimental model for studies of protein structure. PMID- 10669595 TI - Psoralen adducts induced by triplex-forming oligonucleotides are refractory to repair in HeLa cells. AB - The use of triple helix-forming oligonucleotides constitutes an attractive strategy to regulate gene expression by inhibition of transcription. Psoralen oligonucleotide conjugates form, upon irradiation, covalent triplexes and thereby modify the specific target sequence. The processing of such photoproducts on the promoter of the gene coding for the interleukin-2 receptor alpha chain was investigated in HeLa cells and HeLa nuclear extracts. We demonstrate that psoralen cross-links are not repaired within the cell extracts nor inside cells. The mechanism of repair inhibition was elucidated in vitro: the presence of the third strand oligonucleotide inhibits the incision step of the damaged target by repair endonucleases. These results demonstrate the possibility of using this approach to induce a persistent intracellular DNA damage at a specific site and to afford prolonged transcription inhibition. PMID- 10669596 TI - Bacteriophage P22 Abc2 protein binds to RecC increases the 5' strand nicking activity of RecBCD and together with lambda bet, promotes Chi-independent recombination. AB - Bacteriophage P22 Abc2 protein binds to the RecBCD enzyme from Escherichia coli to promote phage growth and recombination. Overproduction of the RecC subunit in vivo, but not RecB or RecD, interfered with Abc2-induced UV sensitization, revealing that RecC is the target for Abc2 in vivo. UV-induced ATP crosslinking experiments revealed that Abc2 protein does not interfere with the binding of ATP to either the RecB or RecD subunits in the absence of DNA, though it partially inhibits RecBCD ATPase activity. Productive growth of phage P22 in wild-type Salmonella typhimurium correlates with the presence of Abc2, but is independent of the absolute level of ATP-dependent nuclease activity, suggesting a qualitative change in the nature of Abc2-modified RecBCD nuclease activity relative to the native enzyme. In lambda phage crosses, Abc2-modified RecBCD could substitute for lambda exonuclease in Red-promoted recombination; lambda Gam could not. In exonuclease assays designed to examine the polarity of digestion, Abc2 protein qualitatively changes the nature of RecBCD double-stranded DNA exonuclease by increasing the rate of digestion of the 5' strand. In this respect, Abc2-modified RecBCD resembles a RecBCD molecule that has encountered the recombination hotspot Chi. However, unlike Chi-modified RecBCD, Abc2-modified RecBCD still possesses 3' exonuclease activity. These results are discussed in terms of a model in which Abc2 converts the RecBCD exonuclease for use in the P22 phage recombination pathway. This mechanism of P22-mediated recombination distinguishes it from phage lambda recombination, in which the phage recombination system (Red) and its anti-RecBCD function (Gam) work independently. PMID- 10669597 TI - Resolution of tethered antiparallel and parallel holliday junctions by the Flp site-specific recombinase. AB - Members of the integrase family site-specific recombinases (also called the tyrosine family) bring about recombination in two steps by exchanging pairs of single strands at a time. The product of the first exchange reaction is a four way DNA junction, the Holliday intermediate. The conformational dynamics by which the recombination complex "isomerizes" from the Holliday-forming to the Holliday resolving mode are not well understood. Experiments with the lambda Int and Escherichia coli XerC/XerD systems imply that the strand configurations at the branch point of the protein-free junction dictate the resolution mode in the protein-bound junction. We have examined the question of strand bias during resolution for the Flp system by using a series of synthetic Holliday junctions that are conformationally constrained by local sequences or by strand tethering. We have not observed a strong resolution bias in favor of the strands designed to assume the "crossed" configuration within the unbound junction. The resolution patterns with antiparallel junctions in a variety of substrate contexts reveal either parity in strand choice, or only modest disparity. On the other hand, the highly biased resolutions observed in the case of tethered parallel junctions can be explained by the non-equivalence in protein occupancy of the DNA arms of these substrates and/or inefficient conversion of cleavage events to recombinants at the tethered ends. PMID- 10669598 TI - Extensive central disruption of a four-way junction on binding CCE1 resolving enzyme. AB - Junction-resolving enzymes are nucleases that are selective for the structure of the four-way DNA junction that is important in genetic recombination. They exhibit selectivity for the structure of the junction, but they also manipulate the structure. Local disruption of DNA structure around the centre of the junction by CCE1 of Saccharomyces cerevisiae has been investigated using 2 aminopurine fluorescence. On binding CCE1, 2-aminopurine bases located at the point of strand exchange exhibit a large increase in fluorescence intensity (up to 39-fold enhancement), consistent with complete unstacking. This was observed for all positions around the centre of the junction, both 5' and 3' to the point of strand exchange. Thymine bases complementary to the modified adenine bases adjacent to the junction centre were strongly reactive to potassium permanganate. The results indicate that binding of CCE1 results in a complete unpairing of the four central base-pairs of the junction, with a lesser disruption of the next base-pairs. PMID- 10669599 TI - Sequence and expression of the kettin gene in Drosophila melanogaster and Caenorhabditis elegans. AB - Kettin is a large modular protein associated with thin filaments in the Z-disc region of insect muscles. The sequence of a 21.3 kb contig of the Drosophila gene has been determined. The corresponding protein sequence has 35 immunoglobulin like (Ig) domains which are separated by shorter linker sequences, except near the N and C termini of the molecule where linker sequences are short or missing. This confirms a model in which each Ig domain binds to an actin protomer. The Drosophila kettin gene is at 62C 1-3 on the third chromosome. Two P-element insertions, l(3)j1D7 and l(3)rL182 are in the kettin gene, and complementation tests showed that existing l(3)dre8 mutations are in the same gene. The RNA was detected in wild-type Drosophila embryos at stage 11, first in the gut invagination region of the mesoderm, and by stage 13 in both visceral and somatic mesoderm. Somatic mesoderm expression became segmental at stage 13. RNA expression was greatly reduced in embryos of P-element homozygotes but normal in heterozygotes. The structure of the flight muscle in all the heterozygous mutants was normal, including the myofibril-cuticle connections, and they were able to fly. Kettin sequence homologous to the Drosophila protein, was identified in the Caenorhabditis elegans genome database. The RNA was detected in pharyngeal, body wall and anal depressor muscles of larvae and adult worms, as well as in the male gonad. Antibody to insect kettin labelled the pharyngeal, body wall, anal depressor and proximal gonadal muscles in adult worms. Body wall muscles were labelled in an obliquely striated pattern consistent with the Z-disc localisation in insect muscle. The relationship of kettin to D-titin, which has been assigned to the same chromosomal locus in Drosophila, is discussed. PMID- 10669600 TI - Direct visualisation of conformational changes in EF(0)F(1) by electron microscopy. AB - The isolated H(+)-ATPase from Escherichia coli (EF(0)F(1)) was investigated by electron microscopy of samples of negatively stained monodisperse molecules, followed by single-particle image processing. The resulting three-dimensional maps showed that the F(1)-part is connected by a prominent stalk to a more peripheral part of F(0). The F(1)-part showed stain-accessible cavities inside. In three-dimensional maps from selected particles, a second stalk could be detected which was thinner than the main stalk and is thought to correspond to the stator.Three-dimensional maps of the enzyme in the absence and in the presence of the substrate analogue adenyl-beta, gamma-imidodiphosphate (AMP-PNP) were calculated. Upon binding of AMP-PNP the three-dimensional maps showed no significant changes in the F(0)-part of EF(0)F(1), whereas a major conformational change in the F(1)-part was observed. (1) The diameter of the F(1)-part decreased upon binding of AMP-PNP mainly in the upper half of F(1). (2) Enzyme particles prepared in the presence of AMP-PNP had a pointed cap at the top of the F(1)-part which was missing in its absence. (3) The stain-accessible cavity inside the F(1) part altered its pattern significantly. PMID- 10669601 TI - Sepia officinalis hemocyanin: A refined 3D structure from field emission gun cryoelectron microscopy. AB - The extracellular respiratory pigment of the cuttlefish Sepia officinalis was observed by cryoelectron microscopy with conventional LaB(6) and field emission gun electron sources at 100 and 200 kV, respectively. Each image series was used to compute one 3D reconstruction volume with correction of the contrast transfer function by Wiener filtering. A strong boosting of the contrast was corrected by band-pass filtering of the final volumes, and a qualitative gain in resolution was observed when using the field emission gun electron microscope. In this volume, a strong signal is present down to 1/18 A(-1) and some meaningful information is obtained down to 1/12.5 A(-1). The complex is composed of five pairs of polypeptide chains and resembles a hollow cylinder with five wall oblique units and five inner arches. Three types of wall-wall connections termed pillar P1 to P3 are visible in this volume and the four functional units present in the arches are each linked to the wall by two arch-wall connections. The dispositions of the functional units in the arches of Sepia and Octopus hemocyanins share no common feature. PMID- 10669602 TI - An EF-hand phage display study of calmodulin subdomain pairing. AB - The interaction between the two EF-hands, EF3 and EF4, in the C-terminal domain of vertebrate calmodulin is addressed using an EF-hand phage display library. Significant specificity is observed in the presence of Ca(2+), as EF3-EF4 heterodimers are favored over EF3-EF3 and EF4-EF4 homodimers. Primarily EF4-type (and not EF3-type) amino acids are selected when an EF3 peptide is used as the target and vice versa. The results show that this specificity is promoted by several factors. There are three positions, corresponding to Phe89, Ala102, and Leu105, that are strongly selected as EF3-type hydrophobic residues with an EF4 target. When EF3 is the target peptide, EF4-type residues, Ile125, Tyr138 and Phe141, are selected. Remarkably, this subset consists of the same three residue positions in EF3 or EF4 and seems to be involved in specifying the heterodimer preference in both cases. In addition, electrostatic repulsion between the acidic monomers in an EF4 homodimer may further influence the preferred stability of heterodimers. This hypothesis is based on the observation that positively charged residues are strongly selected at four positions when EF4 is the target. A survey of EF-hand pairs suggests that charge separation is a common way to achieve efficient attraction of Ca(2+) without causing electrostatic repulsion between the subdomains. No significant specificity of binding is observed in the ion free state or in the presence of magnesium as no sequence is preferentially selected. The residues at the interface between the two EF-hands are thus highly optimized for the Ca(2+) bound state. At some residue positions, EF3-type amino acids are chosen with EF3-target in the presence of Ca(2+). These residues are not involved in the preference for heterodimer over homodimer formation, but represent key positions to mutate in the intact domain to stabilize its Ca(2+)-bound state. PMID- 10669603 TI - High copy display of large proteins on phage for functional selections. AB - We have isolated mutations in the major coat protein P8 of M13 phage that greatly increase the surface display of monomeric or oligomeric proteins. The monomeric protein, human growth hormone (hGH), was fused to the N terminus of P8; libraries of P8 variants were constructed and variants that increased hGH display were selected by binding to the extracellular domain of the hGH receptor. The hGH-P8 fusion protein was found to be extremely tolerant of mutations, and a number of P8 variants were found that increased display to levels that improved detection of the hGH-P8 fusion by almost 100-fold. The increased display likely results from better accommodation of the hGH-P8 fusion protein in the phage coat. Using this high copy display format, it was possible for the first time to detect variants of hGH with very weak affinities for the hGHbp (K(d)>1 microM). The display of a tetrameric protein, streptavidin (approximately 50 kDa), was also increased, suggesting the approach may be general to many proteins. The initial product of a natural or invented selection from a naive library is often a weakly functioning protein. These improvements in high copy display should facilitate the broader goal for selection of proteins with novel functions. PMID- 10669604 TI - Bacteriophage lambda display of complex cDNA libraries: a new approach to functional genomics. AB - We describe the construction and characterization of two lambda surface displayed cDNA expression libraries derived from human brain and mouse embryo. cDNA inserts were obtained by tagged random-priming elongation of commercially available cDNA libraries and cloned into a novel lambda vector at the 3' end of the D capsid protein gene, which produced highly complex repertoires (1x10(8) and 2x10(7) phage). These libraries were affinity selected with a monoclonal antibody against the neural specific factor GAP-43 and with polyclonal antibodies that recognize the EMX1 and EMX2 homeoproteins. In both cases rapid identification of specific clones was achieved, which demonstrates the great potential of the lambda display system for generating affinity selectable cDNA libraries from complex genomes. PMID- 10669605 TI - Structural basis of RXR-DNA interactions. AB - The 9-cis retinoic acid receptor, RXR, binds DNA effectively as a homodimer or as a heterodimer with other nuclear receptors. The DNA-binding sites for these RXR complexes are direct repeats of a consensus sequence separated by one to five base-pairs of intervening space. Here, we report the 2.1 A crystal structure of the RXR-DNA-binding domain as a homodimer in complex with its idealized direct repeat DNA target. The structure shows how a gene-regulatory site can induce conformational changes in a transcription factor that promote homo-cooperative assembly. Specifically, an alpha-helix in the T-box is disrupted to allow efficient DNA-binding and subunit dimerization. RXR displays a relaxed mode of sequence recognition, interacting with only three base-pairs in each hexameric half-site. The structure illustrates how site selection is achieved in this large eukaryotic transcription factor family through discrete protein-protein interactions and the use of tandem DNA binding sites with characteristic spacings. PMID- 10669606 TI - X-ray structure of simian immunodeficiency virus integrase containing the core and C-terminal domain (residues 50-293)--an initial glance of the viral DNA binding platform. AB - The crystal structure of simian immunodeficiency virus (SIV) integrase that contains in a single polypeptide the core and the C-terminal deoxyoligonucleotide binding domain has been determined at 3 A resolution with an R-value of 0.203 in the space group P2(1)2(1)2(1). Four integrase core domains and one C-terminal domain are found to be well defined in the asymmetric unit. The segment extending from residues 114 to 121 assumes the same position as seen in the integrase core domain of avian sarcoma virus as well as human immunodeficiency virus type-1 (HIV 1) crystallized in the absence of sodium cacodylate. The flexible loop in the active site, composed of residues 141-151, remains incompletely defined, but the location of the essential Glu152 residue is unambiguous. The residues from 210 218 that link the core and C-terminal domains can be traced as an extension from the core with a short gap at residues 214-215. The C(alpha) folding of the C terminal domain is similar to the solution structure of this domain from HIV-1 integrase. However, the dimeric form seen in the NMR structure cannot exist as related by the non-crystallographic symmetry in the SIV integrase crystal. The two flexible loops of the C-terminal domain, residues 228-236 and residues 244 249, are much better fixed in the crystal structure than in the NMR structure with the former in the immediate vicinity of the flexible loop of the core domain. The interface between the two domains encompasses a solvent-exclusion area of 1500 A(2). Residues from both domains purportedly involved in DNA binding are narrowly distributed on the same face of the molecule. They include Asp64, Asp116, Glu152 and Lys159 from the core and Arg231, Leu234, Arg262, Arg263 and Lys264 from the C-terminal domain. A model for DNA binding is proposed to bridge the two domains by tethering the 228-236 loop of the C-terminal domain and the flexible loop of the core. PMID- 10669607 TI - Crystal structure of an active two-domain derivative of Rous sarcoma virus integrase. AB - Integration of retroviral cDNA is a necessary step in viral replication. The virally encoded integrase protein and DNA sequences at the ends of the linear viral cDNA are required for this reaction. Previous studies revealed that truncated forms of Rous sarcoma virus integrase containing two of the three protein domains can carry out integration reactions in vitro. Here, we describe the crystal structure at 2.5 A resolution of a fragment of the integrase of Rous sarcoma virus (residues 49-286) containing both the conserved catalytic domain and a modulatory DNA-binding domain (C domain). The catalytic domains form a symmetric dimer, but the C domains associate asymmetrically with each other and together adopt a canted conformation relative to the catalytic domains. A binding path for the viral cDNA is evident spanning both domain surfaces, allowing modeling of the larger integration complexes that are known to be active in vivo. The modeling suggests that formation of an integrase tetramer (a dimer of dimers) is necessary and sufficient for joining both viral cDNA ends at neighboring sites in the target DNA. The observed asymmetric arrangement of C domains suggests that they could form a rotationally symmetric tetramer that may be important for bridging integrase complexes at each cDNA end. PMID- 10669608 TI - Crystal structures of Toxoplasma gondii adenosine kinase reveal a novel catalytic mechanism and prodrug binding. AB - Adenosine kinase (AK) is a key purine metabolic enzyme from the opportunistic parasitic protozoan Toxoplasma gondii and belongs to the family of carbohydrate kinases that includes ribokinase. To understand the catalytic mechanism of AK, we determined the structures of the T. gondii apo AK, AK:adenosine complex and the AK:adenosine:AMP-PCP complex to 2.55 A, 2.50 A and 1.71 A resolution, respectively. These structures reveal a novel catalytic mechanism that involves an adenosine-induced domain rotation of 30 degrees and a newly described anion hole (DTXGAGD), requiring a helix-to-coil conformational change that is induced by ATP binding. Nucleotide binding also evokes a coil-to-helix transition that completes the formation of the ATP binding pocket. A conserved dipeptide, Gly68 Gly69, which is located at the bottom of the adenosine-binding site, functions as the switch for domain rotation. The synergistic structural changes that occur upon substrate binding sequester the adenosine and the ATP gi phosphate from solvent and optimally position the substrates for catalysis. Finally, the 1.84 A resolution structure of an AK:7-iodotubercidin:AMP-PCP complex reveals the basis for the higher affinity binding of this prodrug over adenosine and thus provides a scaffold for the design of new inhibitors and subversive substrates that target the T. gondii AK. PMID- 10669609 TI - Structures of adenylosuccinate synthetase from Triticum aestivum and Arabidopsis thaliana. AB - Catalyzing the first step in the de novo synthesis of adenylmonophosphate, adenylosuccinate synthetase (AdSS) is a known target for herbicides and antibiotics. We have purified and crystallized recombinant AdSS from Arabidopsis thaliana and Tritium aestivum, expressed in Escherichia coli. The structures of A. thaliana and T. aestivum AdSS in complex with GDP were solved at 2.9 A and 3.0 A resolution, respectively. Comparison with the known structures from E. coli reveals that the overall fold is very similar to that of the E. coli protein. The longer N terminus in the plant sequences is at the same place as the longer C terminus of the E. coli sequence in the 3D structure. The GDP-binding sites have one additional hydrogen-bonding partner, which is a plausible explanation for the lower K(m) value. Due to its special position, this partner may also enable GTP to initiate a conformational change, which was, in E. coli AdSS, exclusively activated by ligands at the IMP-binding site. The dimer interfaces show up to six hydrogen bonds and six salt-bridges more than in the E. coli structure, although the contact areas have approximately the same size. PMID- 10669610 TI - Structural basis for the higher Ca(2+)-activation of the regulated actin activated myosin ATPase observed with Dictyostelium/Tetrahymena actin chimeras. AB - Replacement of residues 228-230 or 228-232 of subdomain 4 in Dictyostelium actin with the corresponding Tetrahymena sequence (QTA to KAY replacement: half chimera 1; QTAAS to KAYKE replacement: full chimera) leads to a higher Ca(2+)-activation of the regulated acto-myosin subfragment-1 ATPase activity. The ratio of ATPase activation in the presence of tropomyosin-troponin and Ca(2+) to that without tropomyosin-troponin becomes about four times as large as the ratio for the wild type actin. To understand the structural basis of this higher Ca(2+)-activation, we have determined the crystal structures of the 1:1 complex of Dictyostelium mutant actins (half chimera-1 and full chimera) with gelsolin segment-1 to 2.0 A and 2.4 A resolution, respectively, together with the structure of wild-type actin as a control. Although there were local changes on the surface of the subdomain 4 and the phenolic side-chain of Tyr230 displaced the side-chain of Leu236 from a non-polar pocket to a more solvent-accessible position, the structures of the actin chimeras showed that the mutations in the 228-232 region did not introduce large changes in the overall actin structure. This suggests that residues near position 230 formed part of the tropomyosin binding site on actin in actively contracting muscle. The higher Ca(2+)-activation observed with A230Y-containing mutants can be understood in terms of a three-state model for thin filament regulation in which, in the presence of both Ca(2+) and myosin heads, the local changes of actin generated by the mutation (especially its phenolic side-chain) facilitate the transition of thin filaments from a "closed" state to an "open" state. Between 394 and 469 water molecules were identified in the different structures and it was found that actin recognizes hydrated forms of the adenine base and the Ca ion in the nucleotide binding site. PMID- 10669611 TI - Bacteriophage T4 gene 59 helicase assembly protein binds replication fork DNA. The 1.45 A resolution crystal structure reveals a novel alpha-helical two-domain fold. AB - The bacteriophage T4 gene 59 helicase assembly protein is required for recombination-dependent DNA replication, which is the predominant mode of DNA replication in the late stage of T4 infection. T4 gene 59 helicase assembly protein accelerates the loading of the T4 gene 41 helicase during DNA synthesis by the T4 replication system in vitro. T4 gene 59 helicase assembly protein binds to both T4 gene 41 helicase and T4 gene 32 single-stranded DNA binding protein, and to single and double-stranded DNA. We show here that T4 gene 59 helicase assembly protein binds most tightly to fork DNA substrates, with either single or almost entirely double-stranded arms. Our studies suggest that the helicase assembly protein is responsible for loading T4 gene 41 helicase specifically at replication forks, and that its binding sites for each arm must hold more than six, but not more than 12 nucleotides. The 1.45 A resolution crystal structure of the full-length 217-residue monomeric T4 gene 59 helicase assembly protein reveals a novel alpha-helical bundle fold with two domains of similar size. Surface residues are predominantly basic (pI 9.37) with clusters of acidic residues but exposed hydrophobic residues suggest sites for potential contact with DNA and with other protein molecules. The N-terminal domain has structural similarity to the double-stranded DNA binding domain of rat HMG1A. We propose a speculative model of how the T4 gene 59 helicase assembly protein might bind to fork DNA based on the similarity to HMG1, the location of the basic and hydrophobic regions, and the site size of the fork arms needed for tight fork DNA binding. The fork-binding model suggests putative binding sites for the T4 gene 32 single-stranded DNA binding protein and for the hexameric T4 gene 41 helicase assembly. PMID- 10669612 TI - Crystal structures of an N-terminal fragment from Moloney murine leukemia virus reverse transcriptase complexed with nucleic acid: functional implications for template-primer binding to the fingers domain. AB - Reverse transcriptase (RT) serves as the replicative polymerase for retroviruses by using RNA and DNA-directed DNA polymerase activities coupled with a ribonuclease H activity to synthesize a double-stranded DNA copy of the single stranded RNA genome. In an effort to obtain detailed structural information about nucleic acid interactions with reverse transcriptase, we have determined crystal structures at 2.3 A resolution of an N-terminal fragment from Moloney murine leukemia virus reverse transcriptase complexed to blunt-ended DNA in three distinct lattices. This fragment includes the fingers and palm domains from Moloney murine leukemia virus reverse transcriptase. We have also determined the crystal structure at 3.0 A resolution of the fragment complexed to DNA with a single-stranded template overhang resembling a template-primer substrate. Protein DNA interactions, which are nearly identical in each of the three lattices, involve four conserved residues in the fingers domain, Asp114, Arg116, Asn119 and Gly191. DNA atoms involved in the interactions include the 3'-OH group from the primer strand and minor groove base atoms and sugar atoms from the n-2 and n-3 positions of the template strand, where n is the template base that would pair with an incoming nucleotide. The single-stranded template overhang adopts two different conformations in the asymmetric unit interacting with residues in the beta4-beta5 loop (beta3-beta4 in HIV-1 RT). Our fragment-DNA complexes are distinct from previously reported complexes of DNA bound to HIV-1 RT but related in the types of interactions formed between protein and DNA. In addition, the DNA in all of these complexes is bound in the same cleft of the enzyme. Through site directed mutagenesis, we have substituted residues that are involved in binding DNA in our crystal structures and have characterized the resulting enzymes. We now propose that nucleic acid binding to the fingers domain may play a role in translocation of nucleic acid during processive DNA synthesis and suggest that our complex may represent an intermediate in this process. PMID- 10669613 TI - Solution structure and dynamics of the Rous sarcoma virus capsid protein and comparison with capsid proteins of other retroviruses. AB - The solution structure and dynamics of the recombinant 240 amino acid residue capsid protein from the Rous sarcoma virus has been determined by NMR methods. The structure was determined using 2200 distance restraints and 330 torsion angle restraints, and the dynamics analysis was based on (15)N relaxation parameters (R(1), R(2), and (1)H-(15)N NOE) measured for 153 backbone amide groups. The monomeric protein consists of independently folded N- and C-terminal domains that comprise residues Leu14-Leu146 and Ala150-Gln226, respectively. The domains exhibit different rotational correlation times (16.6(+/-0.1) ns and 12.6(+/-0.1) ns, respectively), are connected by a flexible linker (Ala147-Pro149), and do not give rise to inter-domain NOE values, indicating that they are dynamically independent. Despite limited sequence similarity, the structure of the Rous sarcoma virus capsid protein is similar to the structures determined recently for the capsid proteins of retroviruses belonging to the lentivirus and human T-cell leukemia virus/bovine leukemia virus genera. Structural differences that exist in the C-terminal domain of Rous sarcoma virus capsid relative to the other capsid proteins appear to be related to the occurrence of conserved cysteine residues. Whereas most genera of retroviruses contain a pair of conserved and essential cysteine residues in the C-terminal domain that appear to function by forming an intramolecular disulfide bond during assembly, the Rous sarcoma virus capsid protein does not. Instead, the Rous sarcoma virus capsid protein contains a single cysteine residue that appears to be conserved among the avian C-type retroviruses and is positioned in a manner that might allow the formation of an intermolecular disulfide bond during capsid assembly. PMID- 10669614 TI - Electrostatic dependence of the thrombin-thrombomodulin interaction. AB - The rate constants for the binding interaction between thrombin and a fully active fragment of its anticoagulant cofactor, thrombomodulin, have been determined by surface plasmon resonance. At physiological ionic strength, the k(a) was 6.7x10(6) M(-1) s(-1 )and the dissociation rate constant was 0.033 s( 1). These extremely fast association and dissociation rates resulted in an overall binding equilibrium constant of 4.9 nM, which is similar to previously reported values. Changing the ionic strength from 100 mM to 250 mM NaCl caused a tenfold decrease in the association rate while the dissociation rate did not change significantly. A similar effect was observed with tetramethylammonium chloride. A Debye-Huckel plot of the data had a slope of -6 and an intercept at 0 ionic strength of 10(9) M(-1) s(-1). The same slope and intercept were obtained for data that was collected in the presence of glycerol to slow the association rates. These results show that the thrombin-TM456 interaction is extremely rapid and nearly completely electrostatically steered. An association model is presented in which TM456 approaches thrombin along the direction of the thrombin molecular dipole. PMID- 10669615 TI - Energetics of a strongly pH dependent RNA tertiary structure in a frameshifting pseudoknot. AB - Retroviruses employ -1 translational frameshifting to regulate the relative concentrations of structural and non-structural proteins critical to the viral life cycle. The 1.6 A crystal structure of the -1 frameshifting pseudoknot from beet western yellows virus reveals, in addition to Watson-Crick base-pairing, many loop-stem RNA tertiary structural interactions and a bound Na(+). Investigation of the thermodynamics of unfolding of the beet western yellows virus pseudoknot reveals strongly pH-dependent loop-stem tertiary structural interactions which stabilize the molecule, contributing a net of DeltaH approximately -30 kcal mol(-1) and DeltaG degrees (37) of -3.3 kcal mol(-1) to a total DeltaH and DeltaG degrees (37) of -121 and -16 kcal mol(-1), respectively, at pH 6.0, 0.5 M K(+) by DSC. Characterization of mutant RNAs supports the presence of a C8(+).G12-C26 loop 1-stem 2 base-triple (pK(a)=6.8), protonation of which contributes nearly -3.5 kcal mol(-1) in net stability in the presence of a wild-type loop 2. Substitution of the nucleotides in loop 2 with uridine bases, which would eliminate the minor groove triplex, destroys pseudoknot formation. An examination of the dependence of the monovalent ion and type on melting profiles suggests that tertiary structure unfolding occurs in a manner quantitatively consistent with previous studies on the stabilizing effects of K(+), NH(4)(+) and Na(+) on other simple duplex and pseudoknotted RNAs. PMID- 10669616 TI - Dimerisation mutants of Lac repressor. II. A single amino acid substitution, D278L, changes the specificity of dimerisation. AB - Assembly of the lactose repressor tetramer involves two subunit interfaces, the C terminal heptad repeats, and the monomer-monomer interface. Dimerisation between two monomers of Lac repressor of Escherichia coli lacking the two C-terminal heptad repeats occurs through the interactions between three alpha-helices of each monomer, which form a highly hydrophobic interface. Residues possibly involved in specific dimer formation are known from X-ray studies and from the phenotypes of more than 4000 single amino acid substitutions. During the examination of numerous mutants within the dimerisation interface of Lac repressor, we found that substitution of one amino acid, D278 to leucine, is sufficient to change the specificity of dimerisation. Analysis of this single substitution indicates that D278L mutant Lac repressor represses like wild-type. However, it no longer forms heterodimers with wild-type Lac repressor. PMID- 10669617 TI - Conformational changes in serpins: I. The native and cleaved conformations of alpha(1)-antitrypsin. AB - The serpins (SERine Proteinase INhibitors) are a family of proteins with important physiological roles, including but not limited to the inhibition of chymotrypsin-like serine proteinases. The inhibitory mechan- ism involves a large conformational change known as the S-->R (stressed-->relaxed) transition. The largest structural differences occur in a region around the scissile bond called the reactive centre loop: In the native (S) state, the reactive centre is exposed, and is free to interact with proteinases. In inhibitory serpins, in the cleaved (R) state the reactive centre loop forms an additional strand within the beta-sheet. The latent state is an uncleaved state in which the intact reactive centre loop is integrated into the A sheet as in the cleaved form, to give an alternative R state. The serpin structures illustrate detailed control of conformation within a single protein. Serpins are also an unusual family of proteins in which homologues have native states with different folding topologies. Determination of the structures of inhibitory serpins in multiple conformational states permits a detailed analysis of the mechanism of the S-->R transition, and of the way in which a single sequence can form two stabilised states of different topology. Here we compare the conformations of alpha(1) antitrypsin in native and cleaved states. Many protein conformational changes involve relative motions of large rigid subunits. We determine the rigid subunits of alpha(1)-antitrypsin and analyse the changes in their relative position and orientation. Knowing that the conformational change is initiated by cleavage at the reactive centre, we describe a mechanism of the S-->R transition as a logical sequence of mechanical effects, even though the transition likely proceeds in a concerted manner. PMID- 10669618 TI - Substrate binding site of naphthalene 1,2-dioxygenase: functional implications of indole binding. AB - The three-dimensional structure of the aromatic hydroxylating enzyme naphthalene dioxygenase (NDO) from Pseudomonas sp. NCIB 9816-4 was recently determined. The refinement of the structure together with cyclic averaging showed that in the active site of the enzyme there is electron density for a flat aromatic compound. This compound appears to be an indole adduct, which in Escherichia coli is derived from tryptophan present in the rich culture medium. An indole-dioxygen adduct has been built which fits the electron density convincingly. Support for this interpretation was obtained from crystals of the enzyme purified from cells grown in the absence of tryptophan which had an empty substrate pocket. These types of crystals were soaked in indole solutions and the position of indole in this complex was similar to the corresponding part in the modelled indole-oxygen adduct. This suggests that a peroxide bound to iron end-on attacks the substrate and forms this intermediate. The substrate position has implications for the substrate specificity of the enzyme. Docking studies with indole, naphthalene and biphenyl inside the substrate pocket of NDO suggest the presence of subpockets where the one close to the active site iron is reserved for the binding of the aromatic ring which is hydroxylated upon catalysis. The plausible location for the binding of dioxygen is between this pocket and the catalytic iron. This is in accordance with the enantiospecificity of the products. PMID- 10669619 TI - Active-site gorge and buried water molecules in crystal structures of acetylcholinesterase from Torpedo californica. AB - Buried water molecules and the water molecules in the active-site gorge are analyzed for five crystal structures of acetylcholinesterase from Torpedo californica in the resolution range 2.2-2.5 A (native enzyme, and four inhibitor complexes). A total of 45 buried hydration sites are identified, which are populated with between 36 and 41 water molecules. About half of the buried water is located in a distinct region neighboring the active-site gorge. Most of the buried water molecules are very well conserved among the five structures, and have low displacement parameters, B, of magnitudes similar to those of the main chain atoms of the central beta-sheet structure. The active-site gorge of the native enzyme is filled with over 20 water molecules, which have poor hydrogen bond coordination with an average of 2.9 polar contacts per water molecule. Upon ligand binding, distinct groups of these water molecules are displaced, whereas the others remain in positions similar to those that they occupy in the native enzyme. Possible roles of the buried water molecules are discussed, including their possible action as a lubricant to allow large-amplitude fluctuations of the loop structures forming the gorge wall. Such fluctuations are required to facilitate traffic of substrate, products and water molecules to and from the active-site. Because of their poor coordination, the gorge water molecules can be considered as "activated" as compared to bulk water. This should allow their easy displacement by incoming substrate. The relatively loose packing of the gorge water molecules leaves numerous small voids, and more efficient space-filling by substrates and inhibitors may be a major driving force of ligand binding. PMID- 10669620 TI - ACE gene polymorphism and coronary artery disease: A question of persuasion or statistical confusion? PMID- 10669621 TI - IRS-1 variant: A new risk factor for coronary artery disease? PMID- 10669622 TI - Congenital disorders of platelet signal transduction. PMID- 10669623 TI - Macrophage scavenger receptor class A: A multifunctional receptor in atherosclerosis. AB - In atherogenesis, elevated plasma levels of low density lipoprotein (LDL) lead to the chronic presence of LDL in the arterial wall. There, LDL is modified (eg, oxidized), and these modified lipoproteins activate endothelial cells, which attract circulating monocytes. These monocytes enter the vessel wall, differentiate into macrophages, and subject the modified lipoproteins to endocytosis through scavenger receptor pathways. This unrestricted uptake, which is not limited by intracellular cholesterol levels, eventually leads to the formation of lipid-filled foam cells, the initial step in atherosclerosis. Macrophage scavenger receptor class A (SRA) is thought to be one of the main receptors involved in foam cell formation, mediating the influx of lipids into the macrophages. In addition to this role in modified lipoprotein uptake by macrophages, the SRA has been shown to be important in the inflammatory response in host defense, cellular activation, adhesion, and cell-cell interaction. Given the importance of these processes in atherogenesis, these latter functions may prove to make the SRA a multifunctional player in the atherosclerotic process. PMID- 10669624 TI - Expression of Fas ligand in arteries of hypercholesterolemic rabbits accelerates atherosclerotic lesion formation. AB - Fas ligand (FasL) is expressed by cells of the arterial wall and is present in human atherosclerotic lesions. However, the role of FasL in modifying the initiation and progression of atherosclerosis is unclear. To investigate the role of arterial FasL expression in the development of atherosclerosis, we first established a model of primary lesion formation in rabbit carotid arteries. In this model, infusion of adenoviral vectors into surgically isolated, nondenuded arteries of hypercholesterolemic rabbits leads to the formation of human-like early atherosclerotic lesions. Expression of FasL in arterial endothelium in this model decreased T-cell infiltration and expression of vascular cell adhesion molecule-1 but did not affect expression of intercellular adhesion molecule-1. Intimal lesions grew more rapidly in FasL-transduced arteries than in arteries transduced with a control adenovirus that did not express a transgene. Total intimal macrophage accumulation was increased in FasL-transduced arteries; however, the proportion of lesion area occupied by macrophages was not elevated. The accelerated lesion growth was primarily due to the accumulation of intimal smooth muscle cells with a synthetic proliferative phenotype. There was no significant apoptosis in FasL-transduced or control arteries and no granulocytic infiltrates. Thus, the net result of elevated FasL expression is to accelerate atherosclerotic lesion growth by increasing lesion cellularity. Vascular expression of FasL may contribute to the progression of atherosclerosis. PMID- 10669625 TI - Vascular endothelial cells and smooth muscle cells differ in expression of Fas and Fas ligand and in sensitivity to Fas ligand-induced cell death: implications for vascular disease and therapy. AB - Fas ligand (FasL) is a death factor that induces apoptosis in cells bearing its receptor, Fas. Fas and FasL have been detected in the vessel wall, and it has been proposed that Fas-mediated apoptosis has a role in physiological and pathological cell turnover in the vasculature. Here, we evaluated the expression of Fas in the presence and absence of cytokines on both endothelial cells (ECs) and vascular smooth muscle cells (VSMCs). We also examined the sensitivity of ECs and VSMCs to Fas-mediated apoptosis induced by exposure to multiple Fas agonists: soluble FasL, anti-Fas antibody, and membrane-bound FasL resulting from transduction with a replication-defective adenovirus expressing FasL (Adeno FasL). Cell-surface FasL expression was detected on human ECs with the use of 4 anti-FasL antibodies, whereas cell-surface FasL expression was not detected on VSMCs. Unstimulated ECs expressed relatively low levels of Fas, but expression was upregulated after treatment with tumor necrosis factor-alpha (TNF-alpha) or interferon gamma (IFN-gamma). In contrast, VSMCs expressed relatively high levels of Fas, and treatment with TNF-alpha or IFN-gamma induced little or no upregulation under the conditions of these assays. ECs were resistant to death after exposure to soluble FasL or agonist anti-Fas antibody and also after infection with Adeno-FasL in the presence or absence of cytokine treatment. In contrast, VSMCs remained viable in the presence of soluble FasL or agonist anti Fas antibody, but they underwent apoptosis after infection with Adeno-FasL. IFN gamma enhanced Adeno-FasL-induced death of VSMCs, but TNF-alpha did not. These findings provide insights about the potential role of Fas-mediated apoptosis in the vessel wall and suggest strategies to treat proliferative vascular diseases by exploiting the differential sensitivity of ECs and VSMCs to FasL-induced cell death. PMID- 10669626 TI - Warfarin-induced artery calcification is accelerated by growth and vitamin D. AB - The present studies demonstrate that growth and vitamin D treatment enhance the extent of artery calcification in rats given sufficient doses of Warfarin to inhibit gamma-carboxylation of matrix Gla protein, a calcification inhibitor known to be expressed by smooth muscle cells and macrophages in the artery wall. The first series of experiments examined the influence of age and growth status on artery calcification in Warfarin-treated rats. Treatment for 2 weeks with Warfarin caused massive focal calcification of the artery media in 20-day-old rats and less extensive focal calcification in 42-day-old rats. In contrast, no artery calcification could be detected in 10-month-old adult rats even after 4 weeks of Warfarin treatment. To directly examine the importance of growth to Warfarin-induced artery calcification in animals of the same age, 20-day-old rats were fed for 2 weeks either an ad libitum diet or a 6-g/d restricted diet that maintains weight but prevents growth. Concurrent treatment of both dietary groups with Warfarin produced massive focal calcification of the artery media in the ad libitum-fed rats but no detectable artery calcification in the restricted-diet, growth-inhibited group. Although the explanation for the association between artery calcification and growth status cannot be determined from the present study, there was a relationship between higher serum phosphate and susceptibility to artery calcification, with 30% higher levels of serum phosphate in young, ad libitum-fed rats compared with either of the groups that was resistant to Warfarin-induced artery calcification, ie, the 10-month-old rats and the restricted-diet, growth-inhibited young rats. This observation suggests that increased susceptibility to Warfarin-induced artery calcification could be related to higher serum phosphate levels. The second set of experiments examined the possible synergy between vitamin D and Warfarin in artery calcification. High doses of vitamin D are known to cause calcification of the artery media in as little as 3 to 4 days. High doses of the vitamin K antagonist Warfarin are also known to cause calcification of the artery media, but at treatment times of 2 weeks or longer yet not at 1 week. In the current study, we investigated the synergy between these 2 treatments and found that concurrent Warfarin administration dramatically increased the extent of calcification in the media of vitamin D-treated rats at 3 and 4 days. There was a close parallel between the effect of vitamin D dose on artery calcification and the effect of vitamin D dose on the elevation of serum calcium, which suggests that vitamin D may induce artery calcification through its effect on serum calcium. Because Warfarin treatment had no effect on the elevation in serum calcium produced by vitamin D, the synergy between Warfarin and vitamin D is probably best explained by the hypothesis that Warfarin inhibits the activity of matrix Gla protein as a calcification inhibitor. High levels of matrix Gla protein are found at sites of artery calcification in rats treated with vitamin D plus Warfarin, and chemical analysis showed that the protein that accumulated was indeed not gamma carboxylated. These observations indicate that although the gamma carboxyglutamate residues of matrix Gla protein are apparently required for its function as a calcification inhibitor, they are not required for its accumulation at calcification sites. PMID- 10669627 TI - Lesions in ryanodine channels in smooth muscle cells exposed to oxidized low density lipoprotein. AB - The purpose of the present investigation was to investigate the subcellular basis responsible for the loss of vasoreactivity in atherosclerotic vessels. We have chosen to focus on the potential of oxidized low density lipoprotein (oxLDL), an important atherogenic agent, to alter sarcoplasmic reticulum (SR) structure and function. Vascular smooth muscle cells (VSMCs) were exposed for 1 to 6 days to low concentrations of minimally oxidized LDL. ATP was used to probe SR function in VSMCs. ATP can increase [Ca(2+)](i) in control VSMCs because of a release of Ca(2+) from the SR. However, after chronic exposure to oxLDL, cells lose their ability to increase [Ca(2+)](i) in response to ATP. These cells also exhibit a depressed rise in [Ca(2+)](i) after exposure to ryanodine. These effects were associated with a decreased immunoreactivity for the ryanodine-sensitive Ca(2+) release channels in the SR of oxLDL-treated cells. Immunohistochemical analysis of aortic sections obtained from rabbits fed a cholesterol-supplemented diet revealed a significant decrease in the immunoreactivity for ryanodine channels in the plaque and in the medial layer underlying the plaque. In summary, our data identify oxLDL as a component within the atherosclerotic milieu capable of inducing a decrease in smooth muscle ryanodine channel density. This alteration is associated with a significant defect in the ability of the SR within the smooth muscle cell to regulate Ca(2+). These lesions may contribute to the altered vasoreactivity exhibited by atherosclerotic vessels. PMID- 10669628 TI - Mouse model of femoral artery denudation injury associated with the rapid accumulation of adhesion molecules on the luminal surface and recruitment of neutrophils. AB - Techniques of arterial injury commonly used in animals to mimic endovascular procedures are not suitable for small mouse arteries. This has limited examination of the response to arterial injury in genetically modified mice. We therefore sought to develop a model of transluminal injury to the mouse femoral artery that would be reproducible and result in substantial levels of intimal hyperplasia. Mice of the C57BL/6 strain underwent bilateral femoral artery denudation by passage of an angioplasty guidewire. Intimal hyperplasia was observed in 10% of injured arteries at 1 week, in 88% at 2 weeks, and in 90% at 4 weeks. The mean intimal-to-medial area ratio reached 1.1+/-0.1 at 4 weeks. No intimal proliferation was found in control sham-operated arteries. One hour after injury, the denuded surface was covered with platelets and leukocytes, predominantly neutrophils. This was associated with the accumulation of P selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule 1. Expression of these adhesion molecules was not seen in the underlying medial smooth muscle cells. At 24 hours, few neutrophils remained on the denuded surface. At 1 week, macrophages and platelets were present in the vessel wall, partially covered by regenerated endothelium. Transluminal wire injury to the mouse femoral artery induces abundant intimal hyperplasia formation by 2 and 4 weeks and elicits the rapid accumulation of leukocytes and adhesion molecules on the denuded luminal surface. This model will be a valuable tool to study arterial injury in genetically modified mouse models. PMID- 10669629 TI - Mouse model of transplant arteriosclerosis: role of intercellular adhesion molecule-1. AB - Transplant-accelerated arteriosclerosis in coronary arteries is the major limitation to long-term survival of patients with heart transplantation. The pathogenesis of this disease is not fully understood. Herein, we describe a simplified model of artery allografts in the mouse that allows us to take advantage of transgenic, knockout, or mutant animals. Common carotid arteries or aortic vessels were end-to-end allografted into carotid arteries between C57BL/6J and BALB/c mice. Neointimal lesions were observed as early as 2 weeks after surgery and had progressed at 4 and 6 weeks postoperatively. The lumen of grafted arteries was significantly narrowed due to neointima hyperplasia 4 weeks after transplantation. Using this model, we studied the role of intercellular adhesion molecule-1 (ICAM-1) in the development of transplant arteriosclerosis in ICAM-1 deficient mice. Neointimal lesions of artery grafts from ICAM-1 -/- C57BL/6J to BALB/c mice were reduced up to 60% compared with wild-type controls. MAC-1 (CD11b/18)-positive cells adhering to the surface of ICAM-1 -/- artery grafts were significantly less as identified by en face immunofluorescence, and these positive cells were more abundant in intimal lesions of artery grafts in wild type mice. Furthermore, the major cell component of neointimal lesions 4 weeks after surgery was found to be alpha-actin-positive smooth muscle cells, which were significantly reduced in lesions of ICAM-1 -/- artery grafts. Thus, this model has been proven to be useful for understanding the mechanism of transplant arteriosclerosis. Our findings demonstrate that ICAM-1 is critical in the development of allograft arteriosclerosis via mediation of leukocyte adhesion to, and infiltration into, the vessel wall. PMID- 10669630 TI - Adhesion of monocyte very late antigen-4 to endothelial vascular cell adhesion molecule-1 induces interleukin-1beta-dependent expression of interleukin-6 in endothelial cells. AB - In atheroma, T cell-derived interferon-gamma (INF-gamma) stimulates endothelial cells and facilitates recruitment of monocytes. We investigated potential mechanisms by which these interactions could contribute to local and systemic inflammatory responses. Specifically, we analyzed the expression of interleukin (IL)-1beta and IL-6 in both cell types after coculture, the relevant adhesion molecules in this interaction, and transcriptional control by NF-kappaB. We studied coculture of purified peripheral blood monocytes with human umbilical vein endothelial cells (HUVECs), which were stimulated with INF-gamma (10(6) U/L) to model the activated endothelium of atherosclerotic lesions. Coculture of monocytes with activated HUVECs resulted in release of IL-1beta (40. 6+/-3 pg/24 h, P=0.002) and IL-6 (46.6+/-7 ng/24 h, P=0.0015). Electrophoretic mobility gel shift assay and Northern blotting in each cell type separately revealed NF-kappaB activation in both cell types, IL-1beta mRNA expression predominantly in monocytes, and IL-6 mRNA expression predominantly in HUVECs. The endothelial IL-6 release was IL-1-dependent, because it was suppressed by IL-1 receptor antagonist. Experiments with blocking antibodies demonstrated that binding of monocyte very late antigen-4 (VLA-4) to endothelial vascular cell adhesion molecule-1 (VCAM-1) was necessary for the induction of IL-1beta in monocytes. Binding of monocyte VLA-4 to endothelial VCAM-1 induces NF-kappaB activation in both cell types with expression and release of IL-1beta by monocytes, which in turn stimulates endothelial release of IL-6. The beta(1)-integrin-mediated expression of IL-1beta and IL-6 could contribute to local and systemic inflammatory reactions in atherosclerosis. PMID- 10669631 TI - CD9 participates in endothelial cell migration during in vitro wound repair. AB - CD9, a widely expressed membrane protein of the tetraspanin family, has been implicated in diverse functions, such as signal transduction, cell adhesion, and cell motility. We tested the effects of an anti-CD9 monoclonal antibody (ALMA.1) on the migration and proliferation of human vascular endothelial cells (ECs) during repair of an in vitro mechanical wound mimicking angiogenic processes. ALMA.1 induced dose-dependent inhibition of wound repair with a 35+/-1.5% decrease at 20 microg/mL. Only cell migration was affected, because the rate of proliferation of ECs at the lesion margin was not modified and because the inhibition of repair was also observed for nonproliferating irradiated ECs. Monoclonal antibodies against CD63 tetraspanin (H5C6) and control mouse IgG (MOPC 21) were inactive. CD9, one of the most abundant proteins at the surface of ECs, colocalized with beta(1) or beta(3) integrins on EC membranes in double-labeling immunofluorescence experiments with ALMA.1 and an anti-beta(1) (4B4) or anti beta(3) (SDF.3) monoclonal antibody. Moreover, ALMA.1 and 4B4 had additive inhibitory effects on lesion repair, whereas 4B4 alone also inhibited EC proliferation. In transmembrane Boyden-type assays, ALMA.1 induced dose-dependent inhibition of EC migration toward fibronectin and vitronectin with 45+/-6% and 31+/-10% inhibition, respectively, at 100 microg/mL. 4B4 inhibited migration toward fibronectin at 10 microg/mL but had no effect in the case of vitronectin. Adhesion of ECs to immobilized anti-CD9 monoclonal antibodies induced tyrosine phosphorylated protein levels similar to those observed during interactions with beta(1) or beta(3) integrins. These results point to the involvement of CD9 in EC adhesion and migration during lesion repair and angiogenesis, probably through cooperation with integrins. As such, CD9 is a potential target to inhibit angiogenesis in metastatic and atherosclerotic processes. PMID- 10669632 TI - Insulin-like growth factor binding protein-4 expression is decreased by angiotensin II and thrombin in rat aortic vascular smooth muscle cells. AB - Insulin-like growth factor-I (IGF-I) is a ubiquitous peptide that regulates cellular growth and differentiation and is involved in vascular proliferative responses. The effects of IGF-I are modulated by several IGF-I binding proteins (IGFBPs), including IGFBP-4, the main IGFBP produced by vascular smooth muscle cells (VSMCs). We have previously shown that angiotensin II (Ang II)-induced and thrombin-induced mitogenesis in VSMCs is dependent on autocrine IGF-I. In addition, we have demonstrated that IGF-I and IGFBP-4 mRNA levels are upregulated in the hypertensive aorta of abdominally coarcted rats, a high-renin hypertension model. To obtain further insight into the IGF-I system and to specifically study changes in IGFBP-4, a known inhibitor of IGF-I action, VSMCs were incubated with Ang II or thrombin. Compared with control, Ang II induced an 87+/-2% downregulation of IGFBP-4 mRNA levels at 24 hours, with a 61+/-6% decrease of IGFBP-4 levels, as determined by Western ligand blot analysis. Thrombin had the same depressor effects (87+/-2% for the mRNA levels and 61+/-3% for the protein levels). Ang II and thrombin coincubation with (125)I-IGFBP-4 in the conditioned media failed to reveal any increase in fragmentation, indicating that proteolytic cleavage of IGFBP-4 was not involved in the observed effects. Exogenous recombinant human IGFBP-4 decreased thrombin-induced DNA synthesis of human aortic VSMCs by 64%, whereas anti-IGFBP-4 antibody potentiated thrombin-induced DNA synthesis. These data suggest that downregulation of IGFBP-4 expression in VSMCs may play a critical role in vascular growth response to Ang II and thrombin in normal and diseased states, by increasing the bioavailability of IGF-I for its cell-surface receptor. PMID- 10669633 TI - Zinc finger transcription factor Egr-1 activates Flt-1 gene expression in THP-1 cells on induction for macrophage differentiation. AB - Activation of macrophages is a hallmark of atherosclerosis. Stimulation of human monocytic leukemia THP-1 cells with phorbol 12-myristate 13-acetate (PMA) is known to induce a variety of genes whose function is relevant to activated macrophages. Flt-1, a receptor tyrosine kinase for vascular endothelial growth factor, is expressed in macrophages as well as in endothelial cells and mediates the biological response to vascular endothelial growth factor. In this study, we investigated the molecular mechanisms underlying the inducible expression of the flt-1 gene during the activation of THP-1 cells. Reverse transcription-polymerase chain reaction analysis showed that exposure of THP-1 cells to PMA increases flt 1 mRNA and protein levels. A transfected reporter gene, consisting of the human flt-1 promoter region coupled to the luciferase gene, indicated a direct effect of PMA on transcriptional activity. Transfection analysis of a series of 5' deletion constructs and site-directed mutants localized the PMA-responsive region to a DNA stretch from -174 to -166, which represents overlapping Egr-1/Sp1 transcription factor-binding sites. Competitive gel mobility shift assays and supershift assays showed that PMA induces the binding of Egr-1 to this site. Consistent with these findings, the Egr-1 expression plasmid strongly induced flt 1 promoter activity in a sequence-specific manner. Taken together, our data demonstrate that PMA induces flt-1 gene transcription through an induction of Egr 1 in THP-1 cells, thus providing new evidence that the flt-1 gene is a direct target of Egr-1, the transcription factor primarily induced on macrophage differentiation. PMID- 10669634 TI - Convergence of redox-sensitive and mitogen-activated protein kinase signaling pathways in tumor necrosis factor-alpha-mediated monocyte chemoattractant protein 1 induction in vascular smooth muscle cells. AB - Monocyte chemoattractant protein-1 (MCP-1) is an important component of the inflammatory response of the vessel wall and has been shown to be regulated by cytokines, such as tumor necrosis factor-alpha (TNF-alpha). However, the precise signaling pathways leading to MCP-1 induction have not been fully elucidated in vascular smooth muscle cells (VSMCs). Cytokine signal transduction involves protein kinases as well as reactive oxygen species (ROS). The relation between these 2 factors is not clear. In this study, we show that TNF-alpha induces a parallel phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38MAPK) and increases MCP-1 mRNA expression in cultured VSMCs. Inhibition of ERK1/2 but not p38MAPK caused a partial attenuation of MCP-1 induction (43+/-10% inhibition). Incubation of VSMCs with multiple antioxidants (diphenylene iodonium, liposomal superoxide dismutase, catalase, N-acetylcysteine, dimethylthiourea, and pyrrolidine dithiocarbamate) had no effect on TNF-alpha-mediated MCP-1 upregulation. However, simultaneous blockade of the ERK1/2 and ROS pathways by using PD098059 combined with diphenylene iodonium or N-acetylcysteine potently enhanced the ability of MAPK kinase inhibitors to abrogate MCP-1 mRNA expression (100+/-2% inhibition). Thus, parallel ROS-dependent and ERK1/2-dependent pathways converge to regulate TNF alpha-induced MCP-1 gene expression in VSMCs. These data unmask a complex but organized integration of ROS and protein kinases that mediates cytokine-induced vascular inflammatory gene expression. PMID- 10669635 TI - Inducible expression of manganese superoxide dismutase by phorbol 12-myristate 13 acetate is mediated by Sp1 in endothelial cells. AB - The expression of manganese superoxide dismutase (Mn-SOD), an important component of the cellular defense system against oxidative stress, is induced in response to a variety of stimuli, including cytokines and phorbol esters, in endothelial cells. To define the molecular mechanisms regulating the expression of Mn-SOD, we have characterized the promoter of the human Mn-SOD gene. In calf pulmonary artery endothelial cells, phorbol 12-myristate 13-acetate (PMA) gradually increased Mn-SOD mRNA levels, with a peak at 6 to 12 hours after stimulation. The increase in Mn-SOD mRNA was significantly inhibited by a protein kinase C (PKC) inhibitor (calphostin C) but not by a mitogen-activated protein kinase kinase-1 inhibitor (PD98059) or a p38 mitogen-activated protein kinase inhibitor (SB203580). By reporter gene transfection experiments of a series of promoter deletions and site-directed mutation constructs, we found 2 consensus Sp1 binding sequences located at -97 and at -77 to play an important role in PMA-induced Mn SOD transcription. Electrophoretic gel mobility shift assays have indicated that this sequence serves as an Sp1 binding site. Northern and Western blot analysis has revealed that PMA-induced promoter activity of Mn-SOD correlates with an increased expression of Sp1. Nuclear proteins from PMA-treated calf pulmonary artery endothelial cells displayed an increased DNA binding to the Sp1 site. Furthermore, the Mn-SOD promoter was activated either by overexpression of Sp1 or the constitutively activated form of PKCbeta in an Sp1 site-dependent manner. These results suggest that PMA stimulates transcription of the Mn-SOD gene through an increase in Sp1 expression and thus implicate Sp1 as an effector mediating the PKC-signaling pathway elicited by extracellular signals. PMID- 10669636 TI - Insulin-mediated stimulation of protein kinase Akt: A potent survival signaling cascade for endothelial cells. AB - Insulin exerts potent antiapoptotic effects in neuronal cells and has been suggested to promote angiogenesis. Therefore, we investigated whether insulin inhibits tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Because insulin has been shown to stimulate the protein kinase Akt, we investigated whether activation of Akt contributes to the apoptosis-suppressive effect of insulin and characterized the downstream signaling pathway. Incubation with insulin dose-dependently prevented apoptosis induced by TNF-alpha (50 ng/mL). The extent of apoptosis suppression by insulin was similar to the effect of vascular endothelial growth factor. Pharmacological inhibition of Akt activation or overexpression of a dominant negative Akt mutant prevented the antiapoptotic effect of insulin. Furthermore, we investigated the effect of TNF-alpha on Akt phosphorylation by Western blot analysis with the use of a phosphospecific Akt antibody. Incubation of HUVECs with TNF-alpha induced a marked dephosphorylation of Akt. Insulin counteracted this TNF-alpha-induced dephosphorylation of Akt. Furthermore, we investigated the downstream signaling events. Akt has been shown to mediate its apoptosis suppressive effects via phosphorylation of Bad or caspase-9. However, incubation with insulin did not lead to enhanced phosphorylation of Bad at Ser 136 or Ser 112. In contrast, insulin inhibited caspase-9 activity and prevented caspase-9 induced apoptosis. Mutation of the Akt site within caspase-9 significantly reduced the apoptosis-suppressive effect of insulin. The present study demonstrates an important role for insulin-mediated Akt activation in the prevention of endothelial cell apoptosis, which may importantly contribute to cell homeostasis and the integrity of the endothelium. In endothelial cells, Akt seems to mediate its antiapoptotic effect, at least in part, via phosphorylation of caspase-9 rather than Bad. PMID- 10669637 TI - Expression of tumor necrosis factor-alpha in cultured human endothelial cells stimulated with lipopolysaccharide or interleukin-1alpha. AB - Tumor-necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine with a wide variety of biological effects. The most important source of this cytokine is monocytes/macrophages. It is a potent agonist in the activation of endothelial cells; however, the precise role of endothelial cells as a source of TNF-alpha is not known. In the present study, we addressed the possibility that TNF-alpha is produced by cultured human umbilical vein endothelial cells (HUVEC) stimulated with factors such as lipopolysaccharide (LPS) or interleukin-1alpha (IL-1alpha). LPS and IL-1alpha induced expression of TNF-alpha mRNA in HUVEC. IL-1alpha induced expression and secretion of TNF-alpha protein, but LPS did not induce production of TNF-alpha protein. Most of the TNF-alpha protein in cell lysate was found in the membrane fraction. The mRNA for TNF-alpha-converting enzyme (TACE) was expressed in unstimulated HUVEC, and its level was not altered by treatment with LPS or IL-1alpha. Transfection of HUVEC with full-length cDNA encoding the precursor TNF-alpha enhanced secretion of TNF-alpha protein by these cells, and treatment of the cells with a TACE inhibitor reduced the secretion. These results suggest that HUVEC produce TNF-alpha and have TACE activity. Secreted TNF-alpha may be involved in autocrine activation of endothelial cells, and TNF-alpha retained in cell membrane may serve as a juxtacrine system to activate target cells on the endothelial surface. PMID- 10669638 TI - Fluid flow releases fibroblast growth factor-2 from human aortic smooth muscle cells. AB - This study tested the hypothesis that fluid shear stress regulates the release of fibroblast growth factor (FGF)-2 from human aortic smooth muscle cells. FGF-2 is a potent mitogen that is involved in the response to vascular injury and is expressed in a wide variety of cell types. FGF-2 is found in the cytoplasm of cells and outside cells, where it associates with extracellular proteoglycans. To test the hypothesis that shear stress regulates FGF-2 release, cells were exposed to flow, and FGF-2 amounts were measured from the conditioned medium, pericellular fraction (extracted by heparin treatment), and cell lysate. Results from the present study show that after 15 minutes of shear stress at 25 dyne/cm(2) in a parallel-plate flow system, a small but significant fraction (17%) of the total FGF-2 was released from human aortic smooth muscle cells. FGF 2 levels in the circulating medium increased 10-fold over medium from static controls (P<0.01). A 50% increase in FGF-2 content versus control (P<0.01) was found in the pericellular fraction (extracted by heparin treatment). Furthermore, a significant decrease in FGF-2 was detected in the cell lysate, indicating that FGF-2 was released from inside the cell. Cell permeability studies with fluorescent dextran were performed to examine whether transient membrane disruption caused FGF-2 release. Flow cytometry detected a 50% increase in mean fluorescence of cells exposed to 25 dyne/cm(2) versus control cells. This indicates that the observed FGF-2 release from human aortic smooth muscle cells is likely due to transient membrane disruption on initiation of flow. PMID- 10669639 TI - Homocysteine-induced inhibition of endothelium-dependent relaxation in rabbit aorta: role for superoxide anions. AB - Hyperhomocysteinemia is associated with endothelial dysfunction, although its mechanism is unknown. Isometric tension recordings and lucigenin chemiluminescence were used to assess the effects of homocysteine exposure on endothelium-dependent and -independent relaxation in isolated rabbit aortic rings and superoxide anion (O(2)(-)) production by cultured porcine aortic endothelial cells, respectively. Homocysteine (0.1 to 10 mmol/L) produced a significant (P<0.001) concentration- and time-dependent inhibition of endothelium-dependent relaxation in response to both acetylcholine and the calcium ionophore A23187. Only the intracellular O(2)(-) scavenger 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron, 10 mmol/L) significantly (P<0.001) inhibited the effect of homocysteine on acetylcholine- and A23187-induced relaxation. Incubation of porcine aortic endothelial cells with homocysteine (0.03 to 1 mmol/L for up to 72 hours) caused a significant (P<0.001) time-dependent increase in the O(2)(-) released by these cells on the addition of Triton X-100 (1% [vol/vol]), with levels returning to values comparable to those of control cells at the 72-hour time point. These changes in O(2)(-) levels were associated with a time-dependent increase in endothelial cell superoxide dismutase activity, becoming significant (P<0.001) after 72 hours. Furthermore, the homocysteine-induced increase in endothelial cell O(2)(-) levels was completely inhibited (P<0.001) by the concomitant incubation with either Tiron (10 mmol/L), vitamin C (10 micromol/L), or vitamin E (10 micromol/L). These data suggest that the inhibitory effect of homocysteine on endothelium-dependent relaxation is due to an increase in the endothelial cell intracellular levels of O(2)(-) and provide a possible mechanism for the endothelial dysfunction associated with hyperhomocysteinemia. PMID- 10669640 TI - In vivo evidence of the importance of cardiac angiotensin-converting enzyme in the pathogenesis of cardiac hypertrophy. AB - Cardiac angiotensin-converting enzyme (ACE) may play an important role in regulating cardiac hypertrophy. Angiotensin II (Ang II) stimulates cardiac hypertrophy as well as the production of extracellular matrix. However, it is still unclear whether Ang II exerts a direct effect on cardiac hypertrophy independent of its effect on blood pressure or the circulating renin-angiotensin system. Although ACE inhibitors and/or Ang II receptor antagonists have regressed cardiac hypertrophy, classic pharmacological experiments cannot exclude the contribution of hemodynamics and the circulating renin-angiotensin system. In vivo gene transfer provides the opportunity of assessing the effects of increased cardiac angiotensin in the intact animal without circulating angiotensin or blood pressure. Therefore, we used a "gain of function" approach to obtain local overexpression of cardiac ACE. Transfection of the human ACE vector into rat myocardium resulted in a significant increase in cardiac ACE activity (P<0.01). More interestingly, morphometry at 2 weeks after transfection revealed a significant increase in the thickness and areas of cardiac myocytes in hearts transfected with the ACE vector (P<0.01). In addition, transfection of the ACE vector also resulted in a significant increase in collagen content (P<0.01). This increase in cardiac hypertrophy was abolished by the administration of perindopril. Local transfection of the ACE vector into the heart did not result in systemic effects such as increased blood pressure, heart rate, or serum ACE activity. In summary, we have demonstrated that increased autocrine/paracrine angiotensin can directly cause cardiac hypertrophy independent of systemic factors and hemodynamic effects. This approach has important potentials for defining the role of autocrine/paracrine substances in cardiovascular disease. PMID- 10669641 TI - Complete atherosclerosis regression after human ApoE gene transfer in ApoE deficient/nude mice. AB - The apolipoprotein E (apoE)-deficient mouse is a relevant animal model of human atherosclerosis. Although the prevention of atherosclerosis development has been documented after somatic gene transfer into animal models, regression of lesions remains to be demonstrated. Thus, we used this genetically defined mouse model nn the nude background to show atherosclerosis regression. ApoE-deficient nude mice were infected with 5 x 10(8) or 10(9) plaque-forming units of a first-generation adenovirus encoding human apoE cDNA. The secretion of human apoE resulted in a rapid decrease of total cholesterol, which normalized the hypercholesterolemic phenotype within 14 days (from 600+/-100 to <100 microg/mL). Transgene expression was observed during a period of >4 months, with a normalization of cholesterol and triglyceride levels during 5 months. At that time, we successfully reinjected the recombinant adenovirus and observed the appearance of the human protein as well as the correction of lipoprotein phenotype. In mice killed 6 months-after the first infection, we observed a dose-dependent regression of fatty streak lesions in the aorta. We showed sustained expression of a transgene with a first generation adenoviral vector and a correction of dyslipoproteinemia phenotype leading to lesion regression. These data demonstrate that somatic gene transfer can induce plaque regression. PMID- 10669642 TI - Leptin, peroxisome proliferator-activated receptor-gamma, and CCAAT/enhancer binding protein-alpha mRNA expression in adipose tissue of humans and their relation to cardiovascular risk factors. AB - Obesity is a prevalent disorder that increases the risk for premature cardiovascular disease. The adipose tissue itself plays an active role in the regulation of fuel metabolism and energy homeostasis by expressing a number of regulatory genes, such as leptin, peroxisome proliferator-activated receptor gamma (PPARgamma), and CCAAT/enhancer binding protein-alpha (C/EBPalpha). To study the in vivo relationships among these genes and their associations with cardiovascular risk factors, plasma levels of leptin, lipids, apolipoproteins (apo), insulin, and glucose were measured in 216 obese, 165 nonobese, and 36 weight-losing postobese subjects. mRNA expression of leptin, PPARgamma, and C/EBPalpha in the extraperitoneal and intraperitoneal adipose tissue was quantified in subsets of subjects. In obese individuals, plasma leptin was associated with apoA-I (r=0.2346, P<0.001) and insulin (r=0.2125, P<0.002). Leptin and C/EBPalpha mRNA expression in extraperitoneal and intraperitoneal adipose tissue of obese patients was higher than in the respective tissues of nonobese or postobese subjects. No significant differences among the study groups were found for PPARgamma mRNA expression. Leptin, PPARgamma, and C/EBPalpha mRNA levels correlated with each other in the intraperitoneal and extraperitoneal fat of obese subjects, but multivariate analysis revealed that only C/EBPalpha was a predictor of leptin expression in extraperitoneal tissue (partial r=0.6096, P<0.001). Intraperitoneal PPARgamma expression was inversely related to fasting insulin (r=-0.2888, P<0.017) and a fasting insulin resistance index (r=-0.2814, P<0.021) in obese subjects. In postobese patients, intraperitoneal PPARgamma expression was associated with plasma HDL cholesterol (r=0.5695, P<0.018) and apoA-I (r=0.6216, P<0.008) but was inversely related to LDL cholesterol (r= 0.5101, P<0.03) and apoB (r=-0.6331, P<0.007). These findings suggest a relationship between plasma leptin and HDL metabolism as well as adipose-tissue site-dependent associations among leptin, C/EBP-alpha, and PPAR-gamma mRNA expression. Furthermore, our results suggest that C/EBP-alpha enhances leptin expression in vivo and that PPARgamma mRNA expression is inversely associated with cardiovascular risk factors. PMID- 10669643 TI - LCAT modulates atherogenic plasma lipoproteins and the extent of atherosclerosis only in the presence of normal LDL receptors in transgenic rabbits. AB - Elevated low density lipoprotein cholesterol (LDL-C) and reduced high density lipoprotein cholesterol (HDL-C) concentrations are independent risk factors for coronary heart disease. We have previously demonstrated that overexpression of an enzyme with a well established role in HDL metabolism, lecithin:cholesterol acyltransferase (LCAT), in New Zealand White rabbits not only raises HDL-C concentrations but reduces those of LDL-C as well, ultimately preventing diet induced atherosclerosis. In the present study, the human LCAT gene (hLCAT) was introduced into LDL receptor (LDLr)-deficient (Watanabe heritable hyperlipidemic) rabbits to (1) investigate the role of the LDLr pathway in the hLCAT-mediated reductions of LDL-C and (2) determine the influence of hLCAT overexpression on atherosclerosis susceptibility in an animal model of familial hypercholesterolemia. Heterozygosity or homozygosity for the LDLr defect was determined by polymerase chain reaction, and 3 groups of hLCAT-transgenic (hLCAT+) rabbits that differed in LDLr status were established: (1) LDLr wild type (LDLr+/+), (2) LDLr heterozygotes (LDLr+/-), and (3) LDLr homozygotes (LDLr /-). Data for hLCAT+ rabbits were compared with those of nontransgenic (hLCAT-) rabbits of the same LDLr status. Plasma HDL-C concentrations were significantly elevated in the hLCAT+ animals of each LDLr status. However, LDL-C levels were significantly reduced only in hLCAT+/LDLr+/+ and hLCAT+/LDLr+/- rabbits but not in hLCAT+/LDLr-/- rabbits (405+/-14 versus 392+/-31 mg/dL). Metabolic studies revealed that the fractional catabolic rate (FCR, d(-1)) of LDL apolipoprotein (apo) B-100 was increased in hLCAT+/LDLr+/+ (26+/-4 versus 5+/-0) and hLCAT+/LDLr+/- (4+/-1 versus 1+/-0) rabbits, whereas the FCR of LDL apoB-100 in both groups of LDLr-/- rabbits was nearly identical (0.16+/-0.02 versus 0.15+/ 0.02). Consistently, neither aortic lipid concentrations nor the extent of aortic atherosclerosis was significantly different between hLCAT+/LDLr-/- and hLCAT /LDLr-/- rabbits. Significant correlations were observed between the percent of aortic atherosclerosis and both LDL-C (r=0.985) and LDL apoB-100 FCR (-0.745), as well as between LDL-C and LDL apoB-100 FCR (-0.866). These data are the first to establish that LCAT modulates LDL metabolism via the LDLr pathway, ultimately influencing atherosclerosis susceptibility. Moreover, LCAT's antiatherogenic effect requires only a single functional LDLr allele, identifying LCAT as an attractive gene therapy candidate for the majority of dyslipoproteinemic patients. PMID- 10669644 TI - The Arg123-Tyr166 central domain of human ApoAI is critical for lecithin:cholesterol acyltransferase-induced hyperalphalipoproteinemia and HDL remodeling in transgenic mice. AB - High density lipoprotein (HDL) metabolism and lecithin:cholesterol acyltransferase (LCAT)-induced HDL remodeling were investigated in transgenic mice expressing human apolipoprotein (apo) AI or an apoAI/apoAII chimera in which the Arg123-Tyr166 domain of apoAI was substituted with the Ser12-Ala75 domain of apoAII. Expression of apoAI and of the apoAI/apoAII chimera resulted in a respective 3. 5-fold and 2.9-fold increase of HDL cholesterol. Human LCAT gene transfer into apoAI-transgenic mice resulted in a 5.1-fold increase of endogenous LCAT activity. This increase was associated with a 2. 4-fold increase of the cholesterol ester-to-free cholesterol ratio of HDL, a shift from HDL(3) to HDL(2), and a 2.4-fold increase of HDL cholesterol levels. Agarose gel electrophoresis revealed that human LCAT gene transfer into human apoAI transgenic mice resulted in an increase of pre-beta-HDL and of pre-alpha-HDL. In contrast, human LCAT gene transfer did not affect cholesterol levels and HDL distribution profile in mice expressing the apoAI/apoAII chimera. Mouse LCAT did not "see" a difference between wild-type and mutant human apoAI, whereas human LCAT did, thus localizing the species-specific interaction in the central domain of apoAI. In conclusion, the Arg123-Tyr166 central domain of apoAI is not critical for in vivo lipoprotein association. It is, however, critical for LCAT induced hyperalphalipoproteinemia and HDL remodeling independent of the lipid binding properties of apoAI. PMID- 10669645 TI - Glucosylated glycerophosphoethanolamines are the major LDL glycation products and increase LDL susceptibility to oxidation: evidence of their presence in atherosclerotic lesions. AB - Glycation of both protein and lipid components is believed to be involved in LDL oxidation. However, the relative importance of lipid and protein glycation in the oxidation process has not been established, and products of lipid glycation have not been isolated. Using glucosylated phosphatidylethanolamine (Glc PtdEtn) prepared synthetically, we have identified glycated diacyl and alkenylacyl species among the ethanolamine phospholipids in LDL. Accumulation of these glycation products in LDL incubated with glucose showed a time- and glucose concentration-dependent increase. LDL specifically enriched with Glc PtdEtn (25 nmol/mg protein) showed increased susceptibility to lipid oxidation when dialyzed against a 5-micromol/L Cu(2+) solution. The presence of this glucosylated lipid resulted in a 5-fold increase in production of phospholipid-bound hydroperoxides and 4-fold increase in phospholipid-bound aldehydes. Inclusion of glucosylated phosphatidylethanolamine in the surface lipid monolayer of the LDL resulted in rapid loss of polyunsaturated cholesteryl esters from the interior of the particle during oxidation. Glycated ethanolamine phospholipids were also isolated and identified from atherosclerotic plaques collected from both diabetic and nondiabetic subjects. The present findings provide direct evidence for the previously proposed causative effect of lipid glycation on LDL oxidation. PMID- 10669646 TI - Intravascular ultrasound combined with Raman spectroscopy to localize and quantify cholesterol and calcium salts in atherosclerotic coronary arteries. AB - Coronary intravascular ultrasound (IVUS) can assess arterial wall architecture and localize large intravascular deposits, but it does not provide quantitative chemical information, which is essential in the evaluation of atherosclerotic lesions. Previously, it has been shown that Raman spectroscopy can be used to accurately quantify the relative weights of cholesterol, calcium salts, triglycerides, and phospholipids in homogenized arterial tissue. In the present study, we explore some benefits of combining IVUS and Raman spectroscopy to evaluate the intact arterial wall. IVUS images were collected in vitro from human coronary arterial segments in various stages of disease (n=7). The images were divided into radial segments (11 to 28 per image, 332 in total), each of which was classified visually as calcified or noncalcified tissue. The arteries were opened longitudinally, and Raman spectra were collected from locations at 0. 5-mm intervals across the arterial luminal circumference. The spectra were used to calculate the chemical composition of the arterial wall at the examined locations. Generally, locations containing large amounts of calcium salts, as determined with Raman spectroscopy, were classified as calcified with IVUS. However, small calcific deposits (<6% of weight) were not readily detected with IVUS. The amounts and location of cholesterol determined with Raman spectroscopy were correlated closely with the presence of cholesterol observed by histochemistry, but these deposits could not be located accurately by IVUS. The combination of Raman spectroscopy and IVUS applied in vitro provides detailed information about the amount and location of calcific deposits and lipid pools in atherosclerotic plaques. Future advances in optical fiber technology may allow simultaneous collection of Raman spectra and IVUS images through the same catheter in vivo. PMID- 10669647 TI - ACE gene polymorphism in cardiovascular disease: meta-analyses of small and large studies in whites. AB - The objective of this study was to assess the influence of the ACE gene insertion (I)/deletion (D) polymorphism on plasma ACE activity; blood pressure; and risk of myocardial infarction, ischemic heart disease, and ischemic cerebrovascular disease by comparing small and large studies. The meta-analyses are based on a literature search of MEDLINE up until April 1998 and assessment of bibliographies of published studies and reviews. Forty-six studies were selected, including a total of 32 715 white individuals. Plasma ACE activity was increased 40% and 71% for ID and DD versus II in small studies and 21% and 48% in large studies (small versus large: P<0.001 and P<0.001). Blood pressure was not influenced by genotype. Risk of myocardial infarction and ischemic heart disease was increased by 47% and 29%, respectively, for DD versus ID and II genotypes in small studies but not in large studies (small versus large: P<0.001 for risk of myocardial infarction and P=0.01 for risk of ischemic heart disease). Risk of ischemic cerebrovascular disease was not increased either in the small or in the largest study. In conclusion, the ACE gene polymorphism affects plasma ACE activity but not blood pressure and is not associated with increased risk of myocardial infarction, ischemic heart disease, or ischemic cerebrovascular disease in the largest studies. PMID- 10669648 TI - Homocysteine and lipoprotein(a) interact to increase CAD risk in young men and women. AB - A biochemical link between homocysteine (tHcy) and lipoprotein(a) [Lp(a)] related to fibrin binding has been proposed. This hypothesis has not been specifically examined in human subjects. We sought to determine in a clinical setting whether these risk factors would interact to increase coronary artery disease (CAD) risk. We performed a cross-sectional analysis of 750 men and 403 women referred to a preventive cardiology clinic at the Cleveland Clinic Foundation, in whom baseline tHcy and Lp(a) data were available. Logistic regression after adjusting for standard cardiovascular risk factors was used to estimate the relative risk of CAD in patients with an Lp(a) >/=30 mg/dL and a tHcy >/=17 micromol/L. Neither isolated high tHcy (odds ratio [OR]=1.06, P=0.89) nor isolated high Lp(a) (OR=1.15, P=0.60) appeared to be associated with CAD in women. However, strong evidence of an association was seen when both risk factors were present (OR=4.83, P=0.003). Moreover, this increased risk showed evidence of an interactive effect beyond that attributable to either additive or multiplicative effects of tHcy and Lp(a) (P=0.03). In contrast, both elevated tHcy (OR=1.93, P=0. 05) and elevated Lp(a) (OR=1.87, P=0.01) showed evidence of being independent risk factors for CAD in men. The presence of both risk factors in men did not appear to confer additional risk (OR=2.00, P=0.09), even though ORs as high as 12.4 were observed within specific age intervals. Consistent with prior studies, tHcy and Lp(a) are risk factors, either independently or in concert, for CAD in this clinical population. More significantly, we found evidence that when both risk factors were present in women, the associated risk was greater than what would be expected if the 2 risks were simply acting independently. The absence of such an interactive effect in men may be due to the confounding effects of age manifested as "survivor bias." These clinical findings provide insights into the potential roles of both tHcy and Lp(a) in the pathogenesis of atherosclerosis. PMID- 10669649 TI - Stanol ester margarine alone and with simvastatin lowers serum cholesterol in families with familial hypercholesterolemia caused by the FH-North Karelia mutation. AB - In heterozygous familial hypercholesterolemia (FH), serum low density lipoprotein (LDL) cholesterol levels are already elevated at birth. Premature coronary heart disease occurs in approximately 30% of heterozygous untreated adult patients. Accordingly, to retard development of atherosclerosis, preventive measures for lowering cholesterol should be started even in childhood. To this end, 19 FH families consumed dietary stanol ester for 3 months. Stanol ester margarine lowers the serum cholesterol level by inhibiting cholesterol absorption. Each individual in the study replaced part of his or her daily dietary fat with 25 g of 80% rapeseed oil margarine containing stanol esters (2.24 g/d stanols, mainly sitostanol). The families who consumed this margarine for 12 weeks included 24 children, aged 3 to 13 years, with the North Karelia variant of FH (FH-NK), 4 FH NK parents, and 16 healthy family members, and a separate group of 12 FH-NK adults who consumed the margarine for 6 weeks and who were on simvastatin therapy (20 or 40 mg/d). Fat-soluble vitamins were measured by high-pressure liquid chromatography, and cholesterol precursor sterols (indexes of cholesterol synthesis) and cholestanol and plant sterols (indexes of cholesterol absorption efficiency) were assayed by gas-liquid chromatography. No side effects occurred. Serum LDL cholesterol levels were reduced by 18% (P<0.001), 11%, 12% (P<0.001), and 20% (P<0.001) in the 4 groups, respectively. The serum campesterol-to cholesterol ratios fell by 31% (P<0.001), 29%, 23% (P<0.001), and 36% (P<0.001), respectively, suggesting that cholesterol absorption efficiency was inhibited. Serum lathosterol ratios were elevated by 38% (P<0.001), 11%, 15% (P<0.001), and 19% (P<0.001), respectively, suggesting that cholesterol synthesis was compensatorily upregulated. The FH-NK children increased their serum lathosterol ratio more than did the FH-NK adults treated with stanol ester margarine and simvastatin (P<0.01). In the FH-NK children, serum retinol concentration and alpha-tocopherol-to-cholesterol ratios were unchanged by stanol ester margarine, but alpha- and beta-carotene concentrations and ratios were decreased. As assayed in a genetically defined population of FH patients, a dietary regimen with stanol ester margarine proved to be a safe and effective hypolipidemic treatment for children and adults. In FH-NK adults on simvastatin therapy, serum LDL cholesterol levels could be reduced even further by including a stanol ester margarine in the regimen. PMID- 10669650 TI - New functional promoter polymorphism, CETP/-629, in cholesteryl ester transfer protein (CETP) gene related to CETP mass and high density lipoprotein cholesterol levels: role of Sp1/Sp3 in transcriptional regulation. AB - A new polymorphism located at position -629 (CETP/-629A/C) in the promoter of the cholesteryl ester transfer protein (CETP) gene is described. The -629A allele was associated with lower CETP mass (P<0. 0001) and higher high density lipoprotein cholesterol (P<0.001) than the C allele in a sample of 536 control subjects from the ECTIM study. Transfection studies in HepG2 cells with a luciferase expression vector incorporating a 777-bp fragment of the CETP promoter and containing either A or C at position -629 showed significantly lower luciferase activity with the promoter fragment of the A allele (-25%, P<0.05). By gel-shift assay, DNA-protein interactions were evaluated in nuclear extracts of HepG2 cells with the use of 2 probes (A or C probe) composed of 20 bp of the promoter sequence surrounding the polymorphic site. Two specific complexes of distinct migration rate were identified with the A and the C probe. Competition with an excess of oligonucleotide containing the Sp1 consensus binding site showed that a protein(s) of the Sp transcription factor family was implicated in complex formation with the A probe but not with the C probe. Incubation with specific antibodies indicated that Sp1 and Sp3 bound specifically to the A probe. We introduced mutations in the -629-Sp1 binding site to test its functionality and to define the characteristics of transcription factor binding. We showed, by gel shift assay, that no nuclear proteins bound to the mutated sequence. Transient transfection of HepG2 cells revealed that the expression of the mutated fragment was significantly increased compared with that of the A promoter fragment (25%, P<0.05). The mutated fragment displayed the same activity as that of the C promoter. These results indicate that Sp1 and/or Sp3 repress CETP promoter activity, whereas nuclear factors binding the C allele are without effect on promoter expression. PMID- 10669651 TI - Promoter polymorphisms of human paraoxonase PON1 gene and serum paraoxonase activities and concentrations. AB - Paraoxonase (PON) is a serum enzyme with a wide species distribution. It protects lipoproteins from toxic oxidative modifications and is an antiatherogenic mechanism of major potential. Activity levels of PON are major determinants of the protective function; consequently, factors that influence PON levels are of particular relevance. The present study has identified 3 polymorphisms in the promoter region of the human PON1 gene. Cell transfection studies have revealed their variable impact on promoter activity, with up to 2-fold differences in reporter gene expression. Genotyping studies have established that the polymorphisms are frequent in the population, a finding that is consistent with a major impact on PON concentrations. The physiological relevance of the polymorphisms was underlined by showing that they are associated with highly significant differences in serum concentrations and activities of PON. The study thus firmly establishes a genetic basis for variations in serum PON levels and, consequently, serum PON activity. It is consistent with the suggestion that variations in a major antioxidant function of high density lipoprotein are, to an important degree, genetically determined. PMID- 10669652 TI - Lipoprotein(a) in homozygous familial hypercholesterolemia. AB - Lipoprotein(a) [Lp(a)] is a quantitative genetic trait that in the general population is largely controlled by 1 major locus-the locus for the apolipoprotein(a) [apo(a)] gene. Sibpair studies in families including familial defective apolipoprotein B or familial hypercholesterolemia (FH) heterozygotes have demonstrated that, in addition, mutations in apolipoprotein B and in the LDL receptor (LDL-R) gene may affect Lp(a) plasma concentrations, but this issue is controversial. Here, we have further investigated the influence of mutations in the LDL-R gene on Lp(a) levels by inclusion of FH homozygotes. Sixty-nine members of 22 families with FH were analyzed for mutations in the LDL-R as well as for apo(a) genotypes, apo(a) isoforms, and Lp(a) plasma levels. Twenty-six individuals were found to be homozygous for FH, and 43 were heterozygous for FH. As in our previous analysis, FH heterozygotes had significantly higher Lp(a) than did non-FH individuals from the same population. FH homozygotes with 2 nonfunctional LDL-R alleles had almost 2-fold higher Lp(a) levels than did FH heterozygotes. This increase was not explained by differences in apo(a) allele frequencies. Phenotyping of apo(a) and quantitative analysis of isoforms in family members allowed the assignment of Lp(a) levels to both isoforms in apo(a) heterozygous individuals. Thus, Lp(a) levels associated with apo(a) alleles that were identical by descent could be compared. In the resulting 40 allele pairs, significantly higher Lp(a) levels were detected in association with apo(a) alleles from individuals with 2 defective LDL-R alleles compared with those with only 1 defective allele. This difference of Lp(a) levels between allele pairs was present across the whole size range of apo(a) alleles. Hence, mutations in the LDL-R demonstrate a clear gene-dosage effect on Lp(a) plasma concentrations. PMID- 10669653 TI - Distinct risk profiles of early and advanced atherosclerosis: prospective results from the Bruneck Study. AB - Most epidemiological surveys on risk factors of atherosclerosis were cross sectional in design and did not consider the existence of pathologically distinct processes. The Bruneck Study is a prospective survey in the general community (age range, 40 to 79 years). The baseline examination and first reevaluation were performed in the summers of 1990 and 1995 (participation, 92%; follow-up, 96%). Carotid atherosclerosis was monitored with high-resolution duplex ultrasound. Early (incidence and/or extension of nonstenotic lesions) and advanced (incidence and/or progression of stenosis >40%) stages of atherogenesis were differentiated. The risk profile of early atherogenesis consists of traditional risk factors, such as hypertension, hyperlipidemia, and cigarette smoking (pack-years), supplemented by a variety of less well-established risk conditions, including high body iron stores, hypothyroidism, microalbuminuria, and high alcohol consumption. In contrast, the risk profile of advanced atherogenesis includes markers of enhanced prothrombotic capacity, attenuated fibrinolysis, and clinical conditions known to interfere with coagulation: high fibrinogen, low antithrombin, factor V Leiden mutation, lipoprotein(a) >0.32 g/L, high platelet count, cigarette smoking, and diabetes. Hyperlipidemia and hypertension were of only minor relevance. These findings, along with the epidemiological features of advanced atherogenesis and emergence of an elevated fibrin turnover, suggest atherothrombosis to be a key mechanism in the development of advanced stenotic atherosclerosis. Supplementary 6-category logistic regression models illustrate the changing association between major risk predictors and atherosclerosis of increasing severity and substantiate appropriateness of the 40% threshold applied for the definition of advanced stenotic atherosclerosis. Atherosclerosis is a heterogeneous process that subsumes etiologically and epidemiologically distinct disease entities. The multifactorial etiology of atherosclerosis, which goes far beyond the traditional risk factors, has not yet achieved adequate attention in clinical practice and disease prevention. PMID- 10669654 TI - Insulin resistance syndrome predicts the risk of coronary heart disease and stroke in healthy middle-aged men: the 22-year follow-up results of the Helsinki Policemen Study. AB - The interpretation of conventional multivariate analyses concerning the relation of insulin to the risk of atherosclerotic disease is complex because of correlations of insulin with other risk factors. Therefore, we applied factor analysis to study the clustering of risk factors in the baseline data of the Helsinki Policemen Study (970 healthy men aged 34 to 64 years) and investigated whether these clusterings predict coronary heart disease (CHD) and stroke risk. Areas under the glucose and insulin response curves (AUC glucose and AUC insulin) were used to reflect glucose and insulin levels during oral glucose tolerance tests. During the 22-year follow-up, 164 men had a CHD event, and 70 men had a stroke. Factor analysis of 10 risk factor variables produced 3 underlying factors: insulin resistance factor (comprising body mass index, subscapular skinfold, AUC insulin, AUC glucose, maximal O(2) uptake, mean blood pressure, and triglycerides), lipid factor (cholesterol and triglycerides), and lifestyle factor (physical activity and smoking). In multivariate Cox models, the age adjusted hazard ratio for insulin resistance factor during the 22-year follow-up was 1.28 (95% CI 1.10 to 1.50) with regard to CHD risk and 1.64 (95% CI 1.29 to 2.08) with regard to stroke risk. Lipid factor predicted the risk of CHD but not that of stroke, and lifestyle factor predicted a reduced CHD risk. Factor analysis including only 6 risk factor variables proposed to be central components of insulin resistance syndrome (body mass index, subscapular skinfold, AUC insulin, AUC glucose, mean blood pressure, and triglycerides) produced only a single insulin resistance factor that predicted the risk of CHD and stroke independently of other risk factors. PMID- 10669655 TI - Insulin therapy improves endothelial function in type 2 diabetes. AB - A total of 75 in vivo endothelial function tests (intrabrachial artery infusions of endothelium-dependent [acetylcholine] and -independent [sodium nitroprusside] vasoactive agents) were performed in 18 type 2 diabetic patients (aged 58+/-2 years, body mass index 28.5+/-0.6 kg/m(2), and fasting plasma glucose 229+/-11 mg/dL) and 27 matched normal subjects. These tests were performed before and 6 months after combination therapy with insulin and metformin and before and 6 months after metformin therapy only. Before insulin therapy, blood flow responses to acetylcholine (15 microg/min) were significantly blunted in type 2 diabetic patients (7.5+/-0.7 mL x dL(-1) x min(-1)) compared with normal subjects (11.6+/ 0.9 mL x dL(-1) x min(-1), P<0.01). During insulin therapy, the acetylcholine response increased by 44% to 10.8+/-1.6 mL x dL(-1) x min(-1) (P<0.05). Insulin therapy also significantly increased the blood flow responses to both low and high doses of sodium nitroprusside. We conclude that insulin therapy improves endothelium-dependent and -independent vasodilatation. These data support the idea that insulin therapy has beneficial rather than harmful effects on vascular function. PMID- 10669656 TI - HMG CoA reductase inhibitors reduce plasminogen activator inhibitor-1 expression by human vascular smooth muscle and endothelial cells. AB - The clinical benefit of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) may derive from a qualitative, functional change in atherosclerotic lesions in addition to their lipid-lowering properties. We examined whether statins altered expression of the major determinants of fibrinolytic balance, plasminogen activator inhibitor-1 (PAI-1), and tissue-type plasminogen activator (tPA) in human vascular smooth muscle (SMC) and endothelial (EC) cells. Simvastatin reduced levels of PAI-1 antigen released from SMCs and ECs stimulated with platelet-derived growth factor or transforming growth factor beta (IC(50) approximately 1 micromol/L). Levels of EC-derived tPA increased 2 fold over the same concentrations of simvastatin that inhibited release of PAI-1. Simvastatin's inhibitory effect was mimicked by C3 exoenzyme and prevented by geranylgeranyl pyrophosphate, but not by farnesyl pyrophosphate, suggesting the involvement of geranylgeranyl-modified intermediates. Decreased PAI-1 antigen was correlated with reduced mRNA transcription and activity of the PAI-1 promoter. By inhibiting expression of PAI-1 from SMCs and ECs while increasing expression of tPA from ECs, simvastatin may alter the local fibrinolytic balance within the vessel wall toward increased fibrinolytic capacity that, in turn, would reduce thrombotic risk after plaque rupture. PMID- 10669657 TI - Interaction of anti-phospholipid antibodies with late endosomes of human endothelial cells. AB - Anti-phospholipid antibodies (APLAs) are associated with thrombosis and/or recurrent pregnancy loss. APLAs bind to anionic phospholipids directly or indirectly via a cofactor such as beta(2)-glycoprotein 1 (beta(2)GPI). The lipid target of APLA is not yet established. Recently, we observed that APLAs in vitro can bind lysobisphosphatidic acid (LBPA). The internal membranes of late endosomes are enriched in this phospholipid. The current study was undertaken to determine to what extent binding of APLA to LBPA is correlated with binding to cardiolipin and to beta(2)GPI and to determine whether patient antibodies interact with late endosomes of human umbilical vein endothelial cells (HUVECs) and thus modify the intracellular trafficking of proteins. Binding of patient immunoglobulin G (n=37) to LBPA was correlated significantly with binding to cardiolipin. Although LBPA binding was correlated to a lesser extent with beta(2)GPI binding, we observed that beta(2)GPI binds with high affinity to LBPA. Immunofluorescence studies showed that late endosomes of HUVECs contain LBPA. Patient but not control antibodies recognized late endosomes, but not cardiolipin rich mitochondria, even when we used antibodies that were immunopurified on cardiolipin. Incubation of HUVECs with patient plasma samples immunoreactive toward LBPA resulted in an accumulation of the antibodies in late endosomes and led to a redistribution of the insulinlike growth factor 2/mannose-6-phosphate receptor from the Golgi apparatus to late endosomes. Our results suggest that LBPA is an important lipid target of APLA in HUVECs. These antibodies are internalized by the cells and accumulate in late endosomes. By modifying the intracellular trafficking of proteins, APLA could contribute to several of the proposed pathogenic mechanisms leading to the antiphospholipid syndrome. PMID- 10669658 TI - Effect of individual plasma lipoprotein(a) variations in vivo on its competition with plasminogen for fibrin and cell binding: An in vitro study using plasma from children with idiopathic nephrotic syndrome. AB - Simultaneous natural changes in lipoprotein(a) [Lp(a)] and plasminogen occur in the nephrotic syndrome and offer a unique opportunity to investigate their effects on plasminogen activation under conditions fashioned in vivo. Plasminogen, Lp(a), and apolipoprotein(a) in plasma were characterized, and their competitive binding to carboxy-terminal lysine residues of fibrin and cell membrane proteins was determined in nephrotic children during a flare-up of the disease (61 cases) and after 6 weeks (33 cases) and 6 months (42 cases) of remission. Low plasminogen concentrations (median 1.34 micromol/L, range 0.39 to 1.96 micromol/L) and high Lp(a) levels (median 0.27 g/L, range 0.07 to 2. 57 g/L) were detected at flare-up. These changes were associated with an increased Lp(a) binding ratio onto fibrin (3.13+/-0.48) and cells (1.53+/-0.24) compared with binding ratios of control children (1.31+/-0.19 and 1.05+/-0.07, respectively) with normal plasminogen and low Lp(a) (median 0.071 g/L). After 6 weeks and 6 months of remission, the values for net decrease in Lp(a) binding to fibrin were 1.7+/-0.22 (after 6 weeks) and 1.88+/-0.38 (after 6 months) and were correlated with low Lp(a) concentrations (median 0.2 g/L, range 0.07 to 0.8 g/L; and median 0.12 g/L, range 0.07 to 1.34 g/L) and inversely associated with increased plasminogen levels (median 1.82 micromol/L, range 1.4 to 2.1 micromol/L; and median 1.58 micromol/L, range 1.1 to 2.1 micromol/L). These studies provide the first quantitative evidence that binding of Lp(a) to lysine residues of fibrin and cell surfaces is directly related to circulating levels of both plasminogen and Lp(a) and that these glycoproteins may interact as competitive ligands for these biological surfaces in vivo. This mechanism may be of relevance to the atherothrombotic role of Lp(a), particularly in nephrotic patients. PMID- 10669659 TI - Protective effect of a thrombin receptor (protease-activated receptor 1) gene polymorphism toward venous thromboembolism. AB - The human protease-activated receptor 1 (PAR-1) is activated by thrombin at the surface of platelets and endothelial cells, 2 cells that are implicated in hemostasis and thrombosis. We studied the PAR-1 gene in a large case-control study from the Paris Thrombosis Study (PATHROS), and the possible implication of polymorphisms in venous thromboembolism was evaluated. Two polymorphisms were found in the 5' regulatory region. The first is a C to T transition that is 1426 nucleotides upstream from the translation start site (-1426 C/T), and the second is a 13-bp insertion repeating the preceding -506 5'-CGGCCGCGGGAAG-3' sequence ( 506 I/D, where I indicates insertion and D indicates deletion), a putative cis acting element of the Ets family. The third polymorphism is an A to T transversion in the intervening sequence (IVS) that is 14 nucleotides upstream from the exon 2 start site (IVS-14 A/T). The distribution of the 3 polymorphisms was otherwise similar in the 250 cases and the 1214 controls. A noteworthy sex heterogeneity led us to analyze men and women separately with regard to the -506 I/D polymorphism. We found that allele I was less frequent in male cases than in male controls (0.154 versus 0.247, P<0.01), with an odds ratio at 0.52 (95% CI 0. 32 to 0.82, P<0.01). Furthermore, a reduction of prothrombin fragment 1+2 levels was observed in homozygous carriers of allele -506 I (P=0.04). Altogether, these data suggested a protective effect in men of -506 I/D polymorphism for venous thromboembolism. PMID- 10669660 TI - Factor VII gene polymorphism, factor VII levels, and prevalent cardiovascular disease: the Framingham Heart Study. AB - Elevated factor VII levels have been associated with increased cardiovascular risk in some studies. The arginine/glutamine (Arg/Gln) polymorphism of the factor VII gene has been previously shown to modify factor VII levels. However, the presence of a gene/environment interaction on factor VII levels or a link with cardiovascular disease (CVD) remains uncertain. We studied subjects from the Framingham Heart Study to determine (1) the extent to which this genetic polymorphism affects factor VII levels; (2) whether interactions exist between this polymorphism and environmental factors on factor VII levels; and (3) the association between the polymorphism and CVD. Genotype data and factor VII antigen levels were available in 1816 subjects. Factor VII levels differed significantly among genotypes in an additive fashion: Gln homozygous, 82.7+/ 2.5%; heterozygous, 92.2+/-0.7%; and Arg homozygous, 100. 5+/-0.4% (P<0.0001). The polymorphism was the strongest, single predictor of factor VII levels, explaining 7.7% of the total variance of factor VII levels, whereas other traditional risk factors combined explained an additional 11.5% of the variance. There was an interaction (P=0.02) between the genotype and total cholesterol on factor VII levels, such that the correlation coefficient and slope (factor VII level/total cholesterol) were greatest in Gln/Gln subjects. Among 3204 subjects characterized for genotype and CVD, there was no significant relationship between the genotype and CVD (P=0.12). In the Framingham Heart Study, the Arg/Gln polymorphism was significantly associated with factor VII antigen levels. The strength of the association suggests that genetic variation plays an important role in determining factor VII levels. However, despite being associated with factor VII levels, the Arg/Gln polymorphism was not associated with prevalent CVD. PMID- 10669661 TI - Determinants of population changes in fibrinogen and factor VII over 6 years: the Atherosclerosis Risk in Communities (ARIC) Study. AB - Although numerous cross-sectional studies have identified possible determinants of plasma fibrinogen and factor VII levels, few prospective studies exist. We assessed the longitudinal relation of changes in fibrinogen and factor VII over 6 years with changes to other cardiovascular risk factors in a sample of 440 men and 549 women from the Atherosclerosis Risk in Communities (ARIC) study. Fibrinogen increased more in older participants, those with or who developed diabetes, those who at any time smoked, and those whose plasma HDL cholesterol or triglycerides decreased and increased less in female participants who started hormonal replacement therapy. Factor VII coagulant activity increased more in younger participants, women, those who gained greater weight or developed diabetes, those who quit smoking, those in whom plasma triglycerides decreased, and female participants who received hormonal replacement therapy. Thus, our longitudinal data suggest with some exceptions that adverse changes in cardiovascular risk factors are accompanied by increases in plasma levels of fibrinogen and factor VII. PMID- 10669662 TI - Left atrial appendage myopathy: the importance of serial transesophageal assessment in atrial fibrillation. PMID- 10669663 TI - Transesophageal endoscopic ultrasound-guided mediastinal lymph node aspiration: does the end justify the means? PMID- 10669664 TI - Disorders of ventilation : weakness, stiffness, and mobilization. PMID- 10669665 TI - Diagnosis and natural history of pulmonary infections in transplant recipients. PMID- 10669666 TI - Epidural analgesia and cardiac surgery: worth the risk? PMID- 10669667 TI - Unconventional cancer therapies : what We need is rigorous research, not closed minds. PMID- 10669668 TI - Association of follow-up change of left atrial appendage blood flow velocity with spontaneous echo contrast in nonrheumatic atrial fibrillation. AB - STUDY OBJECTIVES: To evaluate the time-related change of left atrial (LA) appendage flow velocity in chronic atrial fibrillation (AF) by follow-up transesophageal echocardiography (TEE) and to investigate its association with the occurrence of LA spontaneous echo contrast. DESIGN: Prospective follow-up study. SETTING: University-based, tertiary referral medical center. PATIENTS: Forty-seven patients with chronic nonrheumatic AF. INTERVENTIONS: All studied patients underwent both a baseline and follow-up TEE during a mean period of 13 +/- 7 months. MEASUREMENTS AND RESULTS: Baseline TEE revealed that LA spontaneous echo contrast was present in 28 patients (group 1) and was absent in 19 patients (group 2). The LA appendage flow velocity profiles at baseline were significantly lower in group 1 than in group 2; on follow-up, the appendage flow velocities decreased significantly in group 2, but were not significantly changed in group 1. Follow-up TEE revealed that spontaneous echo contrast was persistent in all group 1 patients. In group 2, LA spontaneous echo contrast was newly observed in 9 patients (group 2A) but was persistently absent in 10 patients (group 2B). In group 2A, all of the LA appendage flow velocity profiles decreased significantly at the follow-up study. In group 2B, however, only LA appendage inflow velocity integral showed significant decrease on follow-up; there were no significant changes in LA appendage outflow velocity indexes and peak inflow velocity. CONCLUSIONS: LA appendage flow velocity may decrease with time in some patients with AF, and this change is associated with a new occurrence of LA spontaneous echo contrast. For patients without LA spontaneous echo contrast, serial follow up of the LA appendage flow velocity profiles may be useful for predicting future development of spontaneous echo contrast. Once LA spontaneous echo contrast occurs in AF patients, it tends to persist with time and the LA appendage is usually under a persistently low flow state. PMID- 10669669 TI - Left ventricular thrombus and subsequent thromboembolism in patients with severe systolic dysfunction. AB - STUDY OBJECTIVES: To determine the frequency of left ventricular (LV) thrombi by echocardiography and to define the predictors of LV thrombus and subsequent thromboembolism. DESIGN: Retrospective case-control design. SETTING: Single tertiary care center. PATIENTS: Twenty-eight patients with LV thrombus in a consecutive series of 144 patients with severe LV dysfunction and follow-up period for a mean of 27.6 months. MEASUREMENTS AND RESULTS: Thirty-five clinical and echocardiographic variables were evaluated. The mean age of patients with (n = 28) vs patients without (n = 116) LV thrombus was 50.3 +/- 11.0 years vs 54.2 +/- 11.1 years (p = 0.09), with 22 patients (78.6%) and 78 patients (67.2%) being male (p = 0.24), respectively. The mean ejection fraction (EF) for those with vs those without LV thrombus was 17.5 +/- 5.5 vs 20.0 +/- 6.9 (p = 0. 08), with 16 patients (57.1%) and 42 patients (36.2%) having an EF < 20% (p = 0.04), respectively. The groups were similar with respect to other baseline characteristics, comorbid illnesses, and drug therapies other than anticoagulants. All 28 patients with LV thrombus (100%) and 54 of those without LV thrombus (46.6%) were treated with warfarin. Ischemic etiology of the cardiomyopathy (odds ratio, 4.78; 95% confidence interval, 1.51 to 15.11; p = 0.008) and increased LV internal diastolic dimension (LVIDD; odds ratio, 1.10; 95% confidence interval, 1.03 to 1.18; p = 0.004) were found to be independent predictors of thrombus formation. Peripheral embolism occurred in 5 patients (17.9%) vs 13 patients (11.2%) of those with and without LV thrombi, respectively (p = 0.35). Ischemic etiology of the cardiomyopathy (odds ratio, 3.79; 95% confidence interval, 1. 13 to 12.64; p = 0.03) and EF (odds ratio, 0.91; 95% confidence interval, 0.82 to 1.00; p = 0.04) were found to be independent predictors of systemic embolization. The patients with an embolic event suffered a significantly higher mortality (7 of 18 patients; 38.9%) during the follow-up period when compared to those without an embolic event (13 of 126 patients; 10.3%; p < 0.0001). CONCLUSIONS: We conclude that ischemic cardiomyopathy and dilated LV chamber sizes (LVIDD > 60 mm) are independently associated with LV thrombi. A peripheral embolic event is related to poor long-term survival in this patient group. PMID- 10669670 TI - Ventilatory constraints during exercise in patients with chronic heart failure. AB - We examined the degree of ventilatory constraint in patients with a history of chronic heart failure (CHF; n = 11; mean +/- SE age, 62 +/- 4 years; cardiac index [CI], 2.0 +/- 0.1; and ejection fraction [EF], 24 +/- 2%) and in control subjects (CTLS; n = 8; age, 61 +/- 5 years; CI, 2.6 +/- 0.3) by plotting the tidal flow-volume responses to graded exercise in relationship to the maximal flow-volume envelope (MFVL). Inspiratory capacity (IC) maneuvers were performed to follow changes in end-expiratory lung volume (EELV) during exercise, and the degree of expiratory flow limitation was assessed as the percent of the tidal volume (VT) that met or exceeded the expiratory boundary of the MFVL. CHF patients had significantly (p < 0.05) reduced baseline pulmonary function (FVC, 76 +/- 4%; FEV(1), 78 +/- 4% predicted) relative to CTLS (FVC, 99 +/- 4%; FEV(1), 102 +/- 4% predicted). At peak exercise, oxygen consumption (VO(2)) and minute ventilation (V(E)) were lower in CHF patients than in CTLS (VO(2), 17 +/- 2 vs 32 +/- 2 mL/kg/min; VE, 56 +/- 4 vs 82 +/- 6 L/min, respectively), whereas VE/carbon dioxide output was higher (42 +/- 4 vs 29 +/- 5). In CTLS, EELV initially decreased with light exercise, but increased as VE and expiratory flow limitation increased. In contrast, the EELV in patients with CHF remained near residual volume (RV) throughout exercise, despite increasing flow limitation. At peak exercise, IC averaged 91 +/- 3% and 79 +/- 4% (p < 0.05) of the FVC in CHF patients and CTLS, respectively, and flow limitation was present over > 45% of the VT in CHF patients vs < 25% in CTLS (despite the higher VE in CTLS). The least fit and most symptomatic CHF patients demonstrated the lowest EELV, the greatest degree of flow limitation, and a limited response to increased inspired carbon dioxide during exercise, all consistent with VE constraint. We conclude that patients with CHF commonly breathe near RV during exertion and experience expiratory flow limitation. This results in VE constraint and may contribute to exertional intolerance. PMID- 10669671 TI - Characteristics and prognosis of myocardial infarction in patients with normal coronary arteries. AB - STUDY OBJECTIVES: Myocardial infarction with angiographically normal coronary arteries (MINC) is a life-threatening event with many open questions for physicians and patients. There are little data concerning the prognosis for patients with MINC. DESIGN: Retrospective follow-up study. SETTING: Tertiary referral center. PATIENTS: Patients with MINC were investigated and compared to age- and sex-matched control subjects with myocardial infarction due to coronary artery disease (CAD). The patients were examined clinically using stress exercise and hyperventilation tests. Migraine and Raynaud's symptoms were determined by means of a standardized questionnaire. Serum lipoproteins; the seroprevalence of cytomegalovirus, Helicobacter pylori, and Chlamydia pneumoniae infections; and the most frequent causes of thrombophilia were assessed. MEASUREMENTS AND RESULTS: From > 4,300 angiographies that were performed between 1989 and 1996, 21 patients with MINC were identified. The mean +/- SD patient age at the time of myocardial infarction was 42 +/- 7.5 years. When compared to control subjects (n = 21), patients with MINC had fewer risk factors for CAD. In contrast, MINC patients had more frequent febrile reactions prior to myocardial infarction (six patients vs zero patients; p < 0.05), and the migraine score was significantly higher (7.1 +/- 6.3 vs 2.2 +/- 4.1; p < 0.01). The seroprevalence of antibodies against cytomegalovirus, C pneumoniae, and H pylori tended to be higher in patients with MINC and CAD as compared to matched healthy control subjects. Three patients with MINC vs none with CAD had coagulopathy. During follow-up (53 +/- 37 months), no major cardiac event occurred in the MINC group; no patients with MINC vs nine with CAD (p = 0.0001) underwent repeated angiography. CONCLUSION: High migraine score and prior febrile infection together with a lower cardiovascular risk profile are compatible with an inflammatory and a vasomotor component in the pathophysiology of the acute coronary event in MINC patients. The prognosis for these patients is excellent. PMID- 10669672 TI - Role of transesophageal endosonography-guided fine-needle aspiration in the diagnosis of lung cancer. AB - STUDY OBJECTIVE: Bronchoscopic methods fail to diagnose lung cancer in up to 30% of patients. We studied the role of transesophageal endosonography (EUS)-guided fine-needle aspiration (FNA; EUS-FNA) in such patients. DESIGN: Prospective study. The final diagnosis was confirmed by cytology, histology, or clinical follow-up. SETTING: University hospital. PATIENTS: Thirty-five patients (30 male and 5 female; mean age, 60.9 years; range, 34 to 88 years) with suspected lung cancer in whom bronchoscopic methods failed. Patients with a known diagnosis, recurrence of lung cancer, or mediastinal metastasis from an extrathoracic primary were excluded. INTERVENTIONS: EUS and guided FNA of mediastinal lymph nodes. RESULTS: The procedure was uneventful, and material was adequate in all. The final diagnosis by EUS-FNA was malignancy in 25 patients (11 adenocarcinoma, 10 small cell, 3 squamous cell, and 1 lymphoma) and benign disease in 9 patients (5 inflammatory, 2 sarcoidosis, and 2 anthracosis). Another patient with a benign result had signet-ring cell carcinoma diagnosed on pleural fluid cytology (probably false-negative in EUS-FNA). The sensitivity, specificity, accuracy, and positive and negative predictive values were 96, 100, 97, 100, and 90%, respectively. There were no complications. Reviewing the EUS morphology, the nodes were predominantly located in levels 7 and 8 of American Thoracic Society mediastinal lymph node mapping (subcarinal and paraesophageal region). In seven patients, the punctured nodes were < 1 cm (four malignant and three benign), which are difficult to sample by other methods. The malignant nodes had a hypoechoic, homogenous echotexture. CONCLUSIONS: EUS-FNA is a safe, reliable, and accurate method to establish the diagnosis of suspected lung cancer when bronchoscopic methods fail, especially in the presence of small nodes. PMID- 10669673 TI - Decision-tree sensitivity analysis for cost-effectiveness of chest 2-fluoro-2-D [(18)F]fluorodeoxyglucose positron emission tomography in patients with pulmonary nodules (non-small cell lung carcinoma) in Japan. AB - CONTEXT: Recent studies have demonstrated the potential cost-effectiveness of using 2-fluoro-2-D-[(18)F]fluorodeoxyglucose (FDG) positron emission tomography (PET) in the management of non-small cell lung carcinoma (NSCLC), but because of differences in health-care systems, those findings may not hold true in a Japanese hospital. OBJECTIVE: To assess the cost-effectiveness of the chest CT plus chest FDG-PET strategy in Japan. DESIGN: Decision-tree sensitivity analysis based on the two competing strategies of chest CT-alone vs chest CT plus chest FDG-PET. STUDY SELECTION: A simulation of 1,000 patients in whom NSCLC, stage IIIB or less, was suspected was created using baselines of other relevant variables in regard to sensitivity, specificity, mortality, life expectancy, and cost from published data. METHODS: We surveyed the relevant literature for the choice of variables. MAIN OUTCOME MEASURES: Expected marginal cost and expected life expectancy gain for NSCLC patients. RESULTS: The chest CT plus chest FDG-PET strategy yielded an expected life expectancy gain of 0.607 years (7.3 months) per patient, compared with the alternative strategy of chest CT-alone. Using an FDG PET examination cost of 1.0 x 10(5) yen (around $700 US) per study, the cost increment was 2.18 x 10(5) yen/yr/patient. CONCLUSIONS: The chest CT plus chest FDG-PET strategy in patients with NSCLC is unlikely to be cost-effective in Japan. However, patient life expectancy gain would increase as a result of improved staging of NSCLC. These preliminary results should be confirmed by further studies for specific environments. PMID- 10669674 TI - Non-small cell lung cancer in very young and very old patients. AB - STUDY OBJECTIVE: A cancer registry was analyzed to determine if the clinicopathologic characteristics, treatment modalities, and prognosis of non small cell lung cancer (NSCLC) patients < 40 years of age at diagnosis differed from patients > 80 years of age at diagnosis. DESIGN: Retrospective review of patients with NSCLC diagnosed between 1987 and 1996. SETTING: General teaching hospital in Taiwan. PATIENTS: There were 6,048 cases of NSCLC diagnosed during this period. Among them, 127 patients were < 40 years old and 184 patients were > 80 years old. These patients were selected for our study. MEASUREMENTS: Data regarding demographics, presentation symptoms, histology, tumor staging, treatment modality, and survival were obtained from all patients. Pearson's chi(2) test and the Kaplan-Meier method with a log-rank test were used for statistical analysis. RESULTS: We found significantly more female patients (p < 0.001) and adenocarcinoma (p < 0.001) in the younger group, when compared with the older patients. Cough was the most frequent presenting symptom in both age groups, followed by dyspnea, chest pain, and hemoptysis. There was no statistical difference in the severity of the disease in terms of staging between the two age groups. Young patients received surgical intervention more frequently than the aged (p = 0.025). The older patients received only supportive care more frequently (p = 0.011) than the younger patients. Survival was better in young patients, when compared with other patients or aged patients (p < 0.001). CONCLUSIONS: The female sex and adenocarcinoma were predominant in young NSCLC patients, when compared with the older patients. Young NSCLC patients tended to receive more aggressive treatment and had better survival. PMID- 10669675 TI - Final results of phase III trial in regionally advanced unresectable non-small cell lung cancer: Radiation Therapy Oncology Group, Eastern Cooperative Oncology Group, and Southwest Oncology Group. AB - STUDY OBJECTIVES: The purpose of this phase III clinical trial was to test whether chemotherapy followed by radiation therapy resulted in superior survival to either hyperfractionated radiation or standard radiation in surgically unresectable non-small cell lung cancer. DESIGN: Patients were prospectively randomized to 2 months of cisplatin, vinblastine chemotherapy followed by 60 Gy of radiation at 2.0 Gy per fraction or 1.2 Gy per fraction radiation delivered twice daily to a total dose of 69.6 Gy, or 2.0 Gy per fraction of radiation once daily to 60 Gy. Patients were enrolled from January 1989 through January 1992, and followed for a potential minimum period of 5 years. SETTING: This trial was an intergroup National Cancer Institute-funded trial within the Radiation Therapy Oncology Group, the Eastern Cooperative Oncology Group, and the Southwest Oncology Group. PATIENTS: Patients with surgically unresectable non-small cell lung cancer, clinical stage II, IIIA, and IIIB, were required to have a Karnofsky Performance Status of > or = 70 and a weight loss of < 5% for 3 months before study entry. Four hundred ninety patients were registered on trial, of which 458 patients were eligible. CONCLUSION: Overall survival was statistically superior for the patients receiving chemotherapy and radiation vs the other two arms of the study. The twice-daily radiation therapy arm, although better, was not statistically superior in survival for those patients receiving standard radiation. Median survival for standard radiation was 11.4 months; for chemotherapy and irradiation, 13.2 months; and for hyperfractionated irradiation, 12 months. The respective 5-year survivals were 5% for standard radiation therapy, 8% for chemotherapy followed by radiation therapy, and 6% for hyperfractionated irradiation. PMID- 10669676 TI - Serum interleukin-10 levels as a prognostic factor in advanced non-small cell lung cancer patients. AB - STUDY OBJECTIVE: To investigate the prognostic significance of interleukin (IL) 10 serum levels in advanced non-small cell lung cancer (NSCLC) patients. DESIGN: IL-10 serum levels were measured before chemotherapy, on completion of therapy, and at follow-up by means of a commercially available enzyme-linked immunoassay. The results were then analyzed in comparison with other prognostic variables, and a model predicting overall survival (OS) and time to treatment failure (TTF) was finally generated. SETTING: University hospital. PATIENTS: Sixty consecutive patients with TNM stage III or IV NSCLC undergoing conventional platinum-based regimens. RESULTS: Elevated levels of serum IL-10 were found in cancer patients with respect to healthy control subjects (17.7 +/- 4.4 vs 9.2 +/- 1.5 pg/mL, respectively; p < 0.05), with patients with metastatic disease showing significantly higher levels than patients with undisseminated cancer (21.0 +/- 4.2 vs 14.3 +/- 1.2 pg/mL, respectively; p < 0.05). Following completion of treatment, patients were classified as responders if they had achieved either one of the following: complete response, partial response, or stable disease; and nonresponders, in case of progressive disease. Retrospective analysis of basal IL 10 serum levels in these two subgroups showed a significant difference between responders and nonresponders (15.2 +/- 2.2 vs 21.4 +/- 4.2 pg/mL, respectively; p < 0.05). Moreover, a further significant increase in IL-10 serum levels was observed in nonresponders at the end of therapy (21.4 +/- 4.2 vs 26.0 +/- 4.3 pg/mL, prechemotherapy and postchemotherapy, respectively; p < 0.05), whereas values in responders were found to have significantly decreased (15.2 +/- 2.2 vs 14.8 +/- 2.2 pg/mL, prechemotherapy and postchemotherapy, respectively; p < 0.05). Using univariate and multivariate analyses, both OS and TTF were shown to be affected by the mean pathologic levels of IL-10. Stepwise regression analysis identified IL-10 serum level and stage as the prognostic factors related to OS, and IL-10 serum level and performance status as the prognostic factors related to TTF. CONCLUSIONS: In conclusion, this study shows that the measurement of pretreatment IL-10 serum levels is of independent prognostic utility in patients with NSCLC and may be useful for detection of disease progression. PMID- 10669677 TI - Prognostic assessment of 2,361 patients who underwent pulmonary resection for non small cell lung cancer, stage I, II, and IIIA. AB - STUDY OBJECTIVES: Staging and classification in lung cancer are important for both patient management and clinical research. Results of survival after resection in patients with primary non-small cell lung cancer (NSCLC) are analyzed in order to validate recent refinements of the staging system. DESIGN: Retrospective study; period from 1970 to 1992; follow-up > or = 5 years. PATIENTS: A total of 2,361 previously untreated patients who underwent resection for stage I, II, or IIIA primary NSCLC. MEASUREMENTS: Survival was estimated from the date of operation using the Kaplan-Meier survival analysis method. Deaths within 30 days of operation were excluded. Survival comparisons of different surgical-pathologic TNM classification (based on pathologic examination of resected specimens) as well as further discriminative factors were analyzed by log-rank test. RESULTS: Postoperative death occurred in 3.9% of patients. For survival analyses, 2,263 patients were included. The overall 5-year survival was 937/2,263 (41.4%). Five-year survival in stage IA was 255/404 (63%); in stage IB, 367/797 (46%); in stage IIA, 43/83 (52%); in stage IIB, 210/642 (33%); and in stage IIIA, 63/337 (19%). No significant difference in survival was demonstrated between stages IB and IIA. Until 4 years after surgery, age at operation did not influence survival; after 5 years, patients > 65 years old had a significantly lower survival. CONCLUSION: The TNM staging system accurately reflects the prognosis in primary NSCLC, but some stage definitions can be discussed. Despite the fact that the staging system is built on clinical data, the present analysis, which includes postsurgical data, confirms the similar survival of patients with T2N0M0 and T1N1M0. These results also stress the use of two separate substages, especially because these patients are offered surgery when possible. PMID- 10669678 TI - Nosocomial tuberculosis prevention measures among two groups of US hospitals, 1992 to 1996. AB - OBJECTIVE: To compare trends in nosocomial tuberculosis (TB) prevention measures and health-care worker (HCW) tuberculin skin test (TST) conversion of hospitals with HIV-related Pneumocystis carinii pneumonia (PCP) patients and other US hospitals from 1992 through 1996. DESIGN AND SETTING: Surveys in 1992 and 1996 of 38 hospitals with PCP patients in four high-HIV-incidence cities and 136 other US hospitals from the American Hospital Association membership list. PARTICIPANTS: Twenty-seven hospitals with PCP patients and 103 other US hospitals. RESULTS: In 1992, 63% of PCP hospitals and other US hospitals had rooms meeting Centers for Disease Control and Prevention (CDC) criteria (ie, negative air pressure, six or more air exchanges per hour, and air directly vented to the outside) for acid fast bacilli isolation; in 1996, almost 100% had such isolation rooms. Similarly, in 1992, nonfitted surgical masks were used by HCWs at 60% of PCP hospitals and 68% at other US hospitals, while N95 respirators were used at 90% of PCP hospitals and 83% of other US hospitals in 1996. There was a significant decreasing trend in TST conversion rates among HCWs at both PCP and other US hospitals; however, this trend varied among all hospitals. HCWs at PCP hospitals had a higher risk of TST conversion than those at other US hospitals (relative risk, 1.71; p < 0.0001). CONCLUSION: From 1992 through 1996, PCP and other US hospitals have made similar improvements in their nosocomial TB prevention measures and decreased their HCW TST conversion rate. These data show that most hospitals are compliant with CDC TB guidelines even before the enactment of an Occupational Safety and Health Administration TB standard. PMID- 10669679 TI - Bronchoscopic assessment of the evolution of endobronchial tuberculosis. AB - BACKGROUND: We previously classified forms of endobronchial tuberculosis (EBTB) into seven subtypes by bronchoscopic finding: actively caseating, edematous hyperemic, fibrostenotic, tumorous, granular, ulcerative, and nonspecific bronchitic. STUDY OBJECTIVE: To evaluate the value of this classification in predicting the therapeutic outcome of EBTB. DESIGN: A prospective study with serial bronchoscopy performed from the diagnosis of EBTB to the completion of antituberculosis chemotherapy. PARTICIPANTS: Eighty-one patients with biopsy proven EBTB. INTERVENTIONS: Fiberoptic bronchoscopy was done every month until there was no subsequent change in the endobronchial lesions, every 3 months thereafter, and at the end of treatment. RESULTS: Twenty-two of the 34 cases of actively caseating EBTB changed into the fibrostenotic type, and the other 12 healed without sequelae. Seven of the 11 cases of edematous-hyperemic EBTB changed into the fibrostenotic type, and the other 4 healed. Nine of the 11 cases of granular EBTB, 6 cases of nonspecific bronchitic EBTB, and 2 cases of ulcerative EBTB resolved completely. However, the other two cases of granular EBTB changed into the fibrostenotic type. Seven cases of fibrostenotic EBTB did not improve despite antituberculosis chemotherapy. These various changes in bronchoscopic findings occurred within 3 months of treatment. In 10 cases of tumorous EBTB, 7 progressed to the fibrostenotic type. In addition, new lesions appeared in two cases, and the size of the initial lesions increased in another two cases, even at 6 months after treatment. CONCLUSIONS: The therapeutic outcome of each subtype of EBTB can be predicted by follow-up bronchoscopy during the initial 3 months of treatment, with the exception of the tumorous type. In tumorous EBTB, close and long-term follow-up is advisable because the evolution of the lesions during treatment is very complicated and bronchial stenosis may develop at a later time. PMID- 10669680 TI - Frequency of subspecialty physician care for elderly patients with community acquired pneumonia. AB - STUDY OBJECTIVES: Specialty societies have developed practice guidelines for the treatment of community-acquired pneumonia (CAP). To aid in adapting specialty recommendations for a pneumonia practice guideline at Intermountain Health Care, we investigated which physicians care for pneumonia patients in Utah. We wanted to understand who provides pneumonia care so as to appropriately target the guideline and design tools for implementation. DESIGN: Retrospective observational study. SETTING: Inpatient and outpatient multicenter. PATIENTS: The study population comprised 13,919 (16,420 episodes of pneumonia) Utah resident Medicare beneficiaries > or = 65 years of age who had CAP. Nursing home residents were excluded. MEASUREMENTS: We used Health Care Financing Administration billing records from 1993 through 1995 to identify the physicians involved in the care of pneumonia patients by self-designated specialty. We linked patterns of physician involvement to age, sex, residential zip code, 30-day mortality rate, and whether or not the patient was hospitalized. RESULTS: The involvement of a pneumonia specialist was limited to 11.7% of episodes, with involvement of a pulmonary specialist in 10.6%, an infectious disease (ID) specialist in 0.9%, and the involvement of both specialties in 0.2% of episodes. Greater specialty involvement was observed in episodes resulting in pneumonia hospitalization (20.0% vs 8.6%, respectively; p < 0.0001), death (20.5% vs 11.2%, respectively; p < 0.0001), and episodes among patients with urban county residential zip codes (13.7% vs 7.5%, respectively; p < 0.0001). CONCLUSION: Most episodes of pneumonia, including those with serious consequences, are treated by primary care physicians with little or no involvement from pulmonary or ID specialists. It is not known whether greater or lesser specialty physician involvement would change pneumonia costs or clinical outcomes. PMID- 10669681 TI - The radiologic manifestations of Legionnaire's disease. The Ohio Community-Based Pneumonia Incidence Study Group. AB - STUDY OBJECTIVES: To study the serial radiographic manifestations of Legionnaire's disease from the initial presentation on admission to recovery using strict criteria for the diagnosis of infection. MATERIALS AND METHODS: We prospectively studied the chest radiographs of patients hospitalized with a diagnosis of community-acquired pneumonia in Summit County, Ohio between November 1990 and November 1992. Forty-three patients fulfilled strict criteria for legionellosis. The diagnosis of infection was based on the criteria of "definite" diagnosis as defined by the Ohio Community-Based Pneumonia Incidence Study Group report. The criteria included the isolation of the microorganism, the presence of a significant antibody rise, or the presence of Legionella antigen in the urine. RESULTS: Forty of 43 patients had admission radiographs interpreted as compatible with pneumonia. In spite of appropriate antimicrobial therapy, worsening of the infiltrates was found in more than half of the patients within the first week. Twenty-seven patients were observed to have pleural effusion during the course of hospitalization: 10 effusions were found on admission, another 14 developed during the first week, and 3 new effusions were discovered after the first week. Cavitation was found in only one patient. None of the patients had apical involvement. CONCLUSION: This study confirms previous reports using less stringent etiologic diagnosis criteria that chest radiographic findings in Legionnaire's disease are not specific. Even with appropriate therapy, more than half of the patients will have worsening of the infiltrates during the first week. Pleural effusion is common among our patients, and it is frequently detected during the serial radiographic studies during the first week of hospitalization. Chest radiography in Legionnaire's disease is useful only for the monitoring of disease progression and not for diagnostic purposes. In addition, worsening of infiltrates and pleural effusion are seen in more than half of the patients in spite of appropriate therapy and clinical improvement. PMID- 10669682 TI - Miliary coccidioidomycosis in the immunocompetent. AB - BACKGROUND: Miliary coccidioidomycosis indicates hematogenous or lymphatic spread of Coccidioides immitis and is characterized by the development of multiple small granulomas throughout the lungs and other organs. Previous reports have suggested that this disorder occurs almost exclusively in immunocompromised patients, with most patients succumbing to progressive respiratory failure. In this article, we describe the largest series of immunocompetent patients with miliary coccidioidomycosis, define clinical characteristics, and outline important aspects of diagnosis and treatment. MATERIALS AND METHODS: We identified eight patients (five men and three women; age range, 23 to 65 years) with miliary coccidioidomycosis diagnosed at Kern Medical Center (located in an endemic area) from 1990 to 1997. Four of the patients were white, two were African American, and two were Hispanic. A miliary pattern was defined as the presence of discrete 2- to 10-mm lesions diffusely distributed throughout both lung fields, as shown on chest radiograph. Microscopic examination and culture of C immitis from sputum, tissue, or body fluid confirmed diagnosis. Patients with HIV were excluded. RESULTS: These patients constituted approximately 1% of those admitted to our institution for coccidioidomycosis from 1990 to 1997. Four patients had symptoms for < or = 1 week before admission (acute), and four had symptoms for between 5 and 12 weeks (chronic). Four patients demonstrated a miliary pattern on initial chest radiograph, and two of these patients received an initial diagnosis of miliary coccidioidomycosis. Five patients required mechanical ventilation. Arterial blood gas measurements revealed a mean PO(2) of 54.2 +/- 8. 6 mm Hg and a mean PCO(2) of 32.5 +/- 3.2 mm Hg. Five patients developed ARDS. Five patients had extrapulmonary involvement, with the meninges (n = 4) and skin (n = 4) being the most common sites. All patients were treated with fluconazole and/or amphotericin B. Three patients died; all had chronic involvement and received mechanical ventilation. CONCLUSION: We present eight immunocompetent patients with a lower mortality rate and better outcome than previously reported. In our series, miliary coccidioidomycosis manifested as either an acute respiratory illness or an advanced stage of a chronic illness occurring in the context of widespread dissemination. All who died had chronic involvement. Prompt recognition of miliary coccidioidomycosis is crucial, but may be hindered by the large differential diagnosis. Important diagnostic factors include a history of travel through endemic areas, ethnicity, immunologic status, involvement of multiple organ sites, and pronounced hypoxemia not accounted for by the degree of pulmonary involvement seen on chest radiograph. PMID- 10669683 TI - Intrathoracic Kaposi's sarcoma in women with AIDS. AB - STUDY OBJECTIVE: To describe the radiographic features of intrathoracic Kaposi's sarcoma in women with AIDS. SUBJECTS AND METHODS: From 1987 to 1998, we identified seven women with biopsy-proven (n = 4) or autopsy-proven (n = 3) pulmonary Kaposi's sarcoma. Charts were reviewed for HIV risk factors, cutaneous and/or oropharyngeal Kaposi's sarcoma, CD4 cell count, and differential diagnosis of pulmonary disease prior to the diagnosis of pulmonary Kaposi's sarcoma. Chest radiographs (n = 6), chest CT scans (n = 3), and reports of unavailable chest radiograph (n = 1) closest to the time of diagnosis of pulmonary Kaposi's sarcoma were reviewed for the following: nodular and peribronchovascular opacities; thickened interlobular septa; pleural effusions; lymphadenopathy; and radiographic stage. RESULTS: Mean patient age was 33 years (range, 27 to 42 years). HIV risk factors were IV drug use (n = 2), heterosexual contact (n = 3), and both (n = 2). All patients had prior opportunistic infections. The median CD4 cell count was 18 /microL (mean, 63/microL; range, 5 to 210/microL). Cutaneous Kaposi's sarcoma was diagnosed prior to pulmonary Kaposi's sarcoma in four patients, subsequently in two patients, and not identified in one patient. Oropharyngeal Kaposi's sarcoma was diagnosed prior to pulmonary Kaposi's sarcoma in three patients. Only infection was considered in the differential diagnosis of the patients' pulmonary disease in five patients. One patient presented with acute hemoptysis and died, and one patient recently received a diagnosis of pulmonary Karposi's sarcoma at another hospital. Chest radiographic findings were the following: nodular opacities in five of seven patients (71%); peribronchovascular opacities in six of seven patients (86%); thickened interlobular septa in two of seven patients (29%); pleural effusion in three of seven patients (43%); and lymphadenopathy in two of seven patients (29%). Five of seven patients (71%) were determined to be in radiographic stage 3, one patient in stage 1, and one patient in stage 2. CT demonstrated additional lymphadenopathy in three of three patients, thickened interlobular septa in two of three patients, and pleural effusion in one of three patients, but it did not change the staging of disease in any patient. CONCLUSION: Pulmonary Kaposi's sarcoma can cause diffuse lung disease in women with AIDS. The disease is usually mistaken clinically for pulmonary infection. PMID- 10669684 TI - Distribution of alpha(1)-antitrypsin alleles in patients with bronchiectasis. AB - STUDY OBJECTIVES: Bronchiectasis has been reported in a few patients with homozygous alpha(1)-antitrypsin (AAT) deficiency, but the distribution of AAT alleles among bronchiectatic patients is not known. PATIENTS AND DESIGN: Two hundred two patients, 104 men and 98 women, with a mean age (SD) of 63.7 +/- 15.4 years, had bronchiectasis diagnosed by CT scan alone (n = 178), bronchography with or without CT scan (n = 17), or radiography alone (n = 7). AAT phenotypes (classified according to the protease inhibitor [PI] system) were determined by isoelectric focusing in blood samples obtained from all patients. Bronchiectasis was primary in 121 cases and secondary in 81 patients. Allele and phenotype frequencies were compared retrospectively between bronchiectatic patients and healthy blood donors living in the same geographic area. RESULTS: The PI phenotype frequencies among patients were the following: MM, 81.18%; MS, 11.88%; MZ, 3.46%; IZ, 0.49%; IM, 0.49%; SS, 1.48%; SZ, 0.49%; and ZZ, 0.49%. The allelic frequencies among patients were the following: M, 89.1%; S, 7.67%; Z, 2.72%; and I, 0.49%. There was no difference in the distribution of alleles or phenotypes either between patients and control subjects or between patients with secondary and primary bronchiectasis. A significant difference was found between bronchiectatic patients with and without coexisting emphysema (p = 0.028). This difference was caused by an overrepresentation of PI*Z alleles in bronchiectatic patients with coexisting emphysema. CONCLUSIONS: Our results do not support a physiopathologic implication of the AAT genes in the development of bronchiectasis. We suggest that bronchiectasis may be a consequence of emphysema in PI*Z patients rather than a primary effect. PMID- 10669685 TI - Sputum elastase in steady-state bronchiectasis. AB - STUDY OBJECTIVES: To study the correlations between sputum elastase output with clinical and sputum inflammatory and microbial factors in steady-state bronchiectasis. DESIGN: Prospective recruitment of patients with bronchiectasis (17 women; 48.5 +/- 16.5 years old; FEV(1)/FVC, 1.3 +/- 0.6/2.1 +/- 0.9) for assessment of 24-h sputum output of elastase, bacteria, leukocytes, interleukin (IL)-1beta, IL-8, tumor necrosis factor-alpha, and leukotriene B(4). Clinical variables assessed concomitantly included 24-h sputum volume, lung spirometry, number of lung lobes affected by bronchiectasis, and exacerbation frequency. SETTING: Consecutive recruitment of outpatients (n = 30) in steady-state bronchiectasis. MEASUREMENTS AND RESULTS: Twenty-four-hour sputum elastase output correlated with 24-h sputum volume (r = 0.79, p = 0.0001); number of bronchiectatic lung lobes (r = 0.54, p = 0.0026); percent predicted FEV(1) (r = 0. 48, p = 0.0068); percent predicted FVC (r = -0.49, p = 0.001); and leukocyte output (r = 0.75, p = 0.0001). There was no correlation between the sputum output of bacteria with either inflammatory or enzymatic factors (p > 0.05). CONCLUSION: Our data highlight the importance of elastase and the possibility of independent roles for enzymatic, inflammatory, and microbial components in the pathogenesis of bronchiectasis. Further research on novel therapy targeting each of these components should be pursued. PMID- 10669686 TI - The effect of regular salbutamol on lung function and bronchial responsiveness in patients with primary ciliary dyskinesia. AB - STUDY OBJECTIVE: There is growing evidence that regular beta(2)-agonist use in patients with asthma is associated with decreased airway caliber and increased bronchial responsiveness. The aim of this study was to determine whether regular treatment with beta(2)-agonists induces changes in lung function and bronchial responsiveness in patients with primary ciliary dyskinesia. DESIGN: A randomized, double-blind, placebo-controlled, crossover study. PATIENTS: Nineteen children with primary ciliary dyskinesia. INTERVENTIONS: Subjects received inhaled salbutamol or identical placebo (2 x 100 microg qid) for periods of 6 weeks with a wash-out period of 4 weeks. MEASUREMENTS AND RESULTS: FEV(1) was measured before and 3 weeks and 6 weeks after salbutamol or placebo treatment. High-dose methacholine inhalation tests were performed before and 6 weeks after each treatment. The provocative concentration of methacholine producing a 20% fall in FEV(1) (PC(20)) and maximal airway narrowing (MDeltaFFEV(1)) was measured. No significant change in FEV(1) was observed during the salbutamol or placebo periods. No significant differences in the parameters of bronchial responsiveness (PC(20) and MDeltaFFEV(1)) were noted as the result of either salbutamol or placebo treatment. CONCLUSION: Our data have shown that salbutamol, inhaled regularly for 6 weeks, did not cause either a decline in lung function or an increase in bronchial responsiveness in subjects with primary ciliary dyskinesia. PMID- 10669687 TI - Perceptions of asthma by adolescents at home. AB - OBJECTIVES: To test symptom perception in asthma under natural circumstances and to establish relationships between changes in airway obstruction as indicated by wheeze, dyspnea, general sensations, and emotional state. DESIGN: Continuous in vivo monitoring. METHOD: Symptom perception was tested in 30 adolescents with severe, unstable asthma. They were continuously monitored in their homes for 72 h. Symptom perception was defined as the relation between self-reported dyspnea and airway obstruction as evident from audible wheeze. Tracheal sounds were continuously recorded with wireless telemetry for wheeze assessment. Dyspnea was assessed four times per day on a Likert-type 10-point scale, as well as four times randomly after pager remote command. The subjects kept records of use of medication, daily activities, general symptoms, and mood state in a diary. RESULTS: There were nine subjects with one or two wheeze episodes, another three subjects with three or four episodes, and one subject with almost continuous wheeze. The presence of wheeze in general related significantly to a rise (from individual baseline) in dyspnea of > 2.5 scale points. Acute wheeze was the best predictor of a rise in dyspnea, but prolonged wheeze correlated significantly with negative mood and general symptoms. CONCLUSION: Patients with prolonged airway obstruction perceived symptoms less well and were more vulnerable to negative effects of asthma than patients with acute onset airway obstruction. PMID- 10669688 TI - Low-dose budesonide with the addition of an increased dose during exacerbations is effective in long-term asthma control. On behalf of the Italian Study Group. AB - OBJECTIVES: This study was designed to compare the effects of a 6-month treatment with budesonide 100 microg bid (low dose) and 400 microg bid (standard reference dose) in controlling symptoms and lung function in a group of asthmatics with moderate asthma (baseline FEV(1) > or = 50% and < or = 90% of predicted values) previously treated with inhaled beclomethasone dipropionate (500 to 1,000 microg/d). Moreover, we investigated whether or not asthma exacerbations could be treated by a short-term increase in the daily dose of budesonide. METHODS: After a 2-week run-in period and 1-month treatment with a high dose of budesonide (800 microg bid), 213 patients with moderate asthma were assigned to randomized treatments. Daily treatment included budesonide (bid) plus an additional treatment in case of exacerbation (qid for 7 days). Treatments were as follows: budesonide 400 microg plus placebo (group 1); budesonide 100 microg plus budesonide 200 microg (group 2); and budesonide 100 microg plus placebo (group 3). Symptoms and a peak expiratory flow (PEF) diary were recorded and lung function was measured each month. An exacerbation was defined as a decrease in PEF > 30% below baseline values on 2 consecutive days. RESULTS: We found that that 1-month treatment with a high budesonide dose remarkably reduced all asthma symptoms. Moreover, symptoms were under control in all treatment groups throughout the study period. Similarly, lung function improved and remained stable, and no relevant differences between groups were observed. In each treatment group, the majority of patients had no exacerbations. In patients treated with the standard budesonide dose (group 1), the number of exacerbations and days with exacerbations were significantly lower than in group 3 (intention to-treat analysis). Additionally, patients treated with low budesonide dose plus budesonide (group 2) experienced a significantly lower number of exacerbations and days with exacerbations compared to group 3 (per-protocol analysis). CONCLUSIONS: This study demonstrates that when patients with moderate asthma had reached a stable clinical condition with a high dose of budesonide, a low dose of budesonide (200 microg/d) is as effective as the standard dose (800 microg/d) in the control of symptoms and lung function over a period of several months. Furthermore, results showed that the addition of inhaled budesonide (800 microg/d) at onset of an asthmatic exacerbation has a beneficial clinical effect. PMID- 10669689 TI - Mechanism of CO(2) retention in patients with neuromuscular disease. AB - BACKGROUND: In many studies of patients with muscle weakness, chronic hypercapnia has appeared to be out of proportion to the severity of muscle disease, indicating that factors other than muscle weakness are involved in CO(2) retention. In patients with COPD, the unbalanced inspiratory muscle loading-to strength ratio is thought to trigger the signal for the integrated response that leads to rapid and shallow breathing and eventually to chronic hypercapnia. This mechanism, although postulated, has not yet been assessed in patients with muscular dystrophy. SUBJECTS: Twenty consecutive patients (mean age, 47.6 years; range, 23 to 67 years) were studied: 11 patients with limb-girdle dystrophy, 3 with Duchenne muscular dystrophy, 1 with Charcot-Marie-Tooth syndrome, 1 with Becker muscular dystrophy, 1 with myotonic dystrophy, 1 with facioscapulohumeral dystrophy, and 2 with amyotrophic lateral sclerosis, without any respiratory complaints. Seventeen normal subjects matched for age and sex were studied as a control group. METHODS: Routine spirometry and arterial blood gases, maximal inspiratory and expiratory muscle pressures (MIP and MEP, respectively), and pleural pressure during maximal sniff test (Pplsn), were measured. Mechanical characteristics of the lung were assessed by evaluating lung resistance (RL) and dynamic elastance (Eldyn). Eldyn was assessed as absolute value and as percent of Pplsn; Eldyn (%Pplsn) indicates the elastic load per unit of inspiratory muscle force. Breathing pattern was assessed in terms of time (inspiratory time [TI]; respiratory frequency [Rf]) and volume (tidal volume [VT]) components of the respiratory cycle. RESULTS: A rapid shallow breathing pattern, as indicated by a greater Rf/VT ratio and a lower TI, was found in study patients compared to control subjects. Eldyn was greater in study patients, while MIP, MEP, and Pplsn were lower. PaCO(2) inversely related to VT, TI, and Pplsn (p = 0.012, p = 0.019, and p = 0.002, respectively), whereas it was directly related to Rf, Rf/VT, Eldyn, and Eldyn (%Pplsn) (p < 0.004 to p < 0.0001). Also Eldyn (%Pplsn) inversely related to TI, and the latter positively related to VT. In other words, increase in Eldyn (%Pplsn) was associated with decrease in TI, and the latter was associated with lower VT and greater PaCO(2). Mechanical and breathing pattern variables were introduced in a stepwise multiple regression that selected Eldyn (%Pplsn) (p < 0.0001; r(2) = 0.62) as a unique independent predictor of PaCO(2). CONCLUSIONS: The present study shows that in patients with neuromuscular disease, elastic load and respiratory muscle weakness are responsible for a rapid and shallow breathing pattern leading to chronic CO(2) retention. PMID- 10669690 TI - Hypercapnic ventilatory response in patients with and without obstructive sleep apnea: do age, gender, obesity, and daytime PaCO(2) matter? AB - STUDY OBJECTIVE: To evaluate the relationship between obstructive sleep apnea (OSA) and ventilatory responsiveness to carbon dioxide in both men and women. DESIGN: An analysis of 219 patients referred to an university-based sleep center between 1989 to 1994 was conducted (104 with OSA and 115 without OSA; 43 women and 176 men). These patients had spirometry and a daytime hypercapnic ventilatory response (HCVR) test that was corrected to the patient's ability to attain maximal ventilation. Comparisons between OSA and no-OSA groups, as well as between men and women, were made using multivariate modeling techniques. RESULTS: There was no significant difference in the slope of correlated HCVR (cHCVR) between those with and without OSA (1.57 +/- 0.57 vs 1.63 +/- 0.66; p = 0.48). In men, an inverse correlation between daytime PCO(2) and cHCVR was observed in both crude and multivariate analyses (crude beta-coefficient = - 0.04 +/- 0.02, p = 0.02; adjusted beta-coefficient = 0.07 +/- 0.02, p < 0.01). Although age and cHCVR did not share a significant relationship in the crude analysis (crude beta coefficient = - 0.01 +/- 0.01, p = 0.10), with adjustments for confounding variables, a significant inverse relationship between age and cHCVR was observed (beta-coefficient = - 0.02 +/- 0.01, p = 0.04). On the other hand, in women, only body mass index (BMI) was positively correlated with cHCVR (crude beta coefficient = 0.03 +/- 0.01, p = 0.01; adjusted beta-coefficient = 0.04 +/- 0.01, p < 0.01). CONCLUSION: OSA disorder is not associated with a blunted ventilatory chemoresponsiveness to carbon dioxide. Elevated PaCO(2) and older age are significant correlates for a low cHCVR in men. For women only, BMI was associated with cHCVR. These findings suggest that men and women may have different ventilatory control mechanisms. PMID- 10669691 TI - Effects of sleep stage and age on short-term heart rate variability during sleep in healthy infants and children. AB - STUDY DESIGN: Power spectrum analysis of heart rate variability (HRV) is a noninvasive technique that provides a quantitative assessment of cardiovascular neural control. Using this technique, we studied the autonomic nervous system changes induced by sleep in 14 healthy subjects: 7 infants (mean age, 9.40 +/- 2.32 months) and 7 children (mean age, 8.93 +/- 0.65 years) during a standard all night polysomnographic recording. Our primary aim was to assess the effect of sleep stage and age on short-term HRV during sleep in healthy infants and children. Power spectral density was estimated by autoregressive modeling over 250 consecutive R-R intervals. In this study, we mainly considered two spectral components: the high-frequency (HF) component (0.15 to 0.40 Hz), which reflects parasympathetic cardiovascular modulation; and the low-frequency (LF) component (0.04 to 0.15 Hz), generally considered due to both parasympathetic and sympathetic modulation. RESULTS: Heart rate was higher (p < 0.01 in all sleep stages) and total power lower (p < 0. 02) in infants than in children. HF power was higher in children than in infants (p < 0.05). In infants and children, the ratio between LF and HF powers changed with the various sleep stages (p < 0.02 in infants; p < 0.01 in children): it decreased during deep sleep and increased during rapid eye movement sleep. However, it was invariably lower in children than in infants. CONCLUSION: These findings show that the sleep stage and age both significantly influence short-term HRV during sleep in healthy infants and children. Hence, to provide unbiased results, HRV studies investigating the effects of age on autonomic nervous system activity should segment sleep into the five stages. In addition, despite a relatively small study sample, our data confirm greater parasympathetic control during sleep in children than in infants. PMID- 10669692 TI - The effect of respiratory therapist-initiated treatment protocols on patient outcomes and resource utilization. AB - CONTEXT: Physicians frequently prescribe respiratory treatments to hospitalized patients, but the influence of such treatments on clinical outcomes is difficult to assess. OBJECTIVE: To compare the clinical outcomes of patients receiving respiratory treatments managed by respiratory care practitioner (RCP)-directed treatment protocols or physician-directed orders. DESIGN: A single center, quasi randomized, clinical study. SETTING: Three internal medicine firms from an urban teaching hospital. PATIENTS: Six hundred ninety-four consecutive hospitalized non ICU patients ordered to receive respiratory treatments. MAIN OUTCOME MEASURES: Discordant respiratory care orders, respiratory care charges, hospital length of stay, and patient-specific complications. Discordant orders were defined as written orders for respiratory treatments that were not clinically indicated as well as orders omitting treatments that were clinically indicated according to protocol-based treatment algorithms. RESULTS: Firm A patients (n = 239) received RCP-directed treatments and had a statistically lower rate of discordant respiratory care orders (24.3%) as compared with patients receiving physician directed treatments in firms B (n = 205; 58.5%) and C (n = 250; 56.8%; p < 0.001). No statistically significant differences in patient complications were observed. The average number of respiratory treatments and respiratory care charges were statistically less for firm A patients (10.7 +/- 13.7 treatments; $868 +/- 1,519) as compared with patients in firms B (12.4 +/- 12.7 treatments, $1,124 +/- 1,339) and C (12.3 +/- 13.4 treatments, $1, 054 +/- 1,346; p = 0.009 [treatments] and p < 0.001 [respiratory care charges]). CONCLUSIONS: Respiratory care managed by RCP-directed treatment protocols for non-ICU patients is safe and showed greater agreement with institutional treatment algorithms as compared with physician-directed respiratory care. Additionally, the overall utilization of respiratory treatments was significantly less among patients receiving RCP directed respiratory care. PMID- 10669693 TI - Osteoporosis and lung transplantation: a prospective study. AB - STUDY OBJECTIVE: Osteoporosis is a well-recognized complication of lung transplantation that may significantly impair the quality of life of transplant recipients. We performed a prospective study of bone mineral density (BMD) before and after transplantation to determine the degree of bone mass loss associated with lung transplantation Patients and design: We conducted a prospective study of BMD in 28 patients with various end-stage respiratory diseases pretransplantation and 6 to 12 months posttransplantation. The BMD of the lumbar spine (LS) and femoral neck (FN) were measured. All 28 patients were treated only with vitamin D and calcium supplementation posttransplant. The primary endpoint was the percentage change in BMD. The secondary endpoint was the incidence of fractures posttransplant. A univariate analysis was conducted to determine the various risk factors associated with bone mass loss pretransplant and posttransplant. RESULTS: Prior to transplantation, moderate to severe bone disease was evident. The mean (+/- SD) pretransplant T score (the number of SDs from the peak bone mass) and Z score (the number of SDs from the age-matched mean) for the LS were -1.72 +/- 1.37 and -1.44 +/- 1.31, respectively. The mean pretransplant T score and Z score for the FN were -2.65 +/- 1.01 and -1.5 +/- 1.43, respectively. Within 6 to 12 months posttransplant, the mean BMD for the LS decreased by 4.76% (p < 0.001), while the mean BMD for the FN decreased by 5.3% (p < 0.001). Five of the 28 patients (18%) suffered osteoporotic fractures posttransplant, while no fractures were documented pretransplant. The cumulative steroid dose posttransplant was associated with a drop in BMD for the LS and FN (r = 0.39, p = 0.039 and r = 0.63, p < 0.001, respectively), while a negative association was found between cumulative steroid use pretransplant and baseline LS and FN T scores (r = -0.4, p = 0. 02 and r = -0.43, p = 0.023, respectively). CONCLUSION: Within 6 to 12 months after lung transplantation, there is a significant decrease in BMD at both the LS and FN levels (approximately 5%) despite vitamin D and calcium supplementation. This drop in BMD is associated with a relatively high incidence of osteoporotic fractures posttransplant. PMID- 10669694 TI - Mediastinal fibrosis is associated with human leukocyte antigen-A2. AB - OBJECTIVE: To determine the association between mediastinal fibrosis and human leukocyte antigen (HLA) genes. DESIGN: Case-control study. SETTING: Vanderbilt University Medical Center. SUBJECTS: Nineteen consecutive patients with mediastinal fibrosis who presented to the pulmonary clinic at Vanderbilt University Medical Center from 1987 to 1996. The control subjects were 21,086 whites who were cadaveric kidney donors from October 1987 through December 1993. MEASUREMENTS: HLA testing was performed on blood samples from all 19 cases. Information on HLA typing for the control subjects was obtained from the United Network for Organ Sharing. Frequency of HLA class I and II antigens found in the cases was compared with the frequency in the control subjects. RESULTS: The relative risk of mediastinal fibrosis among subjects with the HLA-A2 antigen was 3.32 times that of those who lacked this antigen (95% confidence interval, 1.19 to 9. 2). CONCLUSION: HLA-A2 was strongly associated with mediastinal fibrosis, suggesting that an abnormal immune response is important in the pathogenesis of this disease. Key words: Histoplasma capsulatum; human leukocyte antigen-A2; mediastinal fibrosis PMID- 10669695 TI - Asbestos in extrapulmonary sites: omentum and mesentery. AB - STUDY OBJECTIVES: Asbestos fibers have not been reported in tissues from the peritoneal cavity. Therefore, omentum, mesentery, and lung tissues from 20 individuals in whom mesothelioma was diagnosed were analyzed for asbestos bodies and asbestos fibers. DESIGN: Tissue was digested and prepared filters were analyzed by light microscopy and analytical transmission electron microscopy. RESULTS: Asbestos bodies were found in the lungs of 18 individuals, mesentery samples from 5, and omentum samples from 2. Uncoated asbestos fibers were found in lungs of 19 patients, 17 of whom had fibers in at least one extrapulmonary site. The most common asbestos in the omentum and mesentery was amosite. Several features of asbestos found in lung influenced the likelihood of amphibole fibers being found in the omentum or mesentery. Lung features included total amphibole fiber burden, length, aspect ratio, and ferruginous body burden. An increased total ferruginous body burden was strongly associated with increased likelihood of detecting amphiboles in the omentum (p < 0. 05). CONCLUSION: Asbestos fibers reach areas in the peritoneal cavity where some mesotheliomas develop. This study suggests their presence can be predicted based on concentrations and characteristics of fiber burdens in lung tissue. PMID- 10669696 TI - Etiology and microbial patterns of pulmonary infiltrates in patients with orthotopic liver transplantation. AB - STUDY OBJECTIVE: To evaluate the etiology and microbial patterns of pulmonary infiltrates in liver transplant patients using a bronchoscopic diagnostic approach and the impact of diagnostic results on antimicrobial treatment decisions. DESIGN: A prospective cohort study. SETTING: A 1,000-bed tertiary-care university hospital. PATIENTS AND METHODS: Fifty consecutive liver transplant patients with 60 episodes of pulmonary infiltrates (33 episodes during mechanical ventilation) were studied using flexible bronchoscopy with protected specimen brush (PSB) and BAL. RESULTS: A definite infectious etiology was confirmed in 29 episodes (48%). Eighteen episodes corresponded to probable pneumonia (30%), 10 episodes had noninfectious etiologies (17%), and 3 remained undetermined (5%). Opportunistic infections were the most frequent etiology (16/29, 55%, including 1 mixed etiology). Bacterial infections (mainly Gram-negative) accounted for 14 of 29 episodes (48%), including 1 of mixed etiology. The majority of bacterial pneumonia episodes (n = 10, 71%) occurred in period 1 (1 to 28 days posttransplant) during mechanical ventilation, whereas opportunistic episodes were predominant in periods 2 and 3 (29 to 180 days and > 180 days posttransplant, respectively; n = 14, 82%). Microbial treatment was changed according to diagnostic results in 21 episodes (35%). CONCLUSIONS: Microbial patterns in liver transplant patients with pulmonary infiltrates corresponded to nosocomial, mainly Gram-negative bacterial pneumonia in period 1, and to opportunistic infections in period 2 and, to a lesser extent, period 3. A comprehensive diagnostic evaluation including PSB and BAL fluid examination frequently guided specific antimicrobial therapy. PMID- 10669697 TI - Assessment of prognosis in patients with community-acquired pneumonia who require mechanical ventilation. AB - STUDY OBJECTIVES: Knowing that mortality is high in patients who require mechanical ventilation patients with community-acquired pneumonia (CAP), we hypothesized that the severity of acute lung injury could be used along with nonpulmonary factors to identify patients with the highest risk of death. We formulated a prediction model to quantitate the risk of hospital mortality in this population of patients. DESIGN: Historical prospective study using data collected over the first 24 h of mechanical ventilation. We utilized a hypoxemia index-(1 - lowest [PaO(2)/PAO(2)]) x (minimum fraction of inspired oxygen to maintain PaO(2) at > 60 mm Hg) x 100], where PAO(2) is the alveolar partial pressure of oxygen-to grade the severity of acute lung injury on a scale from 0 to 100. SETTING: Tertiary care university hospital ICU. PATIENTS: One hundred forty-four adult patients mechanically ventilated for respiratory failure caused by CAP. MEASUREMENTS AND RESULTS: Hospital mortality was 46% (n = 66). Multivariate logistic regression analysis revealed five independent predictors of hospital mortality: (1) the extent of lung injury assessed by the hypoxemia index; (2) the number of nonpulmonary organs that failed; (3) immunosuppression; (4) age > 80 years; and (5) medical comorbidity with a prognosis for survival < 5 years. At a 50% mortality threshold, the prediction model correctly classified outcome in 88% of cases. All patients with > 95% predicted probability of death died in hospital. CONCLUSIONS: Based on clinical parameters measured over the first 24 h of mechanical ventilation, this model accurately identified critically ill, mechanically ventilated patients with CAP for whom prolonged intensive care may not be of benefit. PMID- 10669698 TI - Topical antibiotics on tracheostoma prevents exogenous colonization and infection of lower airways in children. AB - INTRODUCTION: Patients requiring long-term ventilation are at high risk of lower airway infections, generally of endogenous development. Patients on long-term ventilation, in particular via a tracheostomy, may develop tracheobronchitis or pneumonia of exogenous pathogenesis, ie, caused by microorganisms not carried in the oropharynx. The frequency of exogenous colonization or infection has previously been reported to be as high as 33%. A prospective observational cohort study of 2 years was undertaken to evaluate the efficacy of topical antibiotics in the prevention of exogenous colonization or infection of the lower airways. The antibiotic combination of polymyxin E and tobramycin in a 2% paste was applied four times a day on the tracheostoma. MATERIALS AND METHODS: A total of 23 children (median age, 4.1 months; range, 0 to 215 months) were enrolled in the study from September 1, 1996, until August 30, 1998. Surveillance samples of the oropharynx were obtained before tracheostomy and thereafter twice weekly. Diagnostic samples of the lower airways were taken once weekly and on clinical indication. RESULTS: Fourteen children (61%) had a total of 16 episodes of tracheal colonization or infection with 20 potentially pathogenic microorganisms. Only one child had tracheobronchitis with Streptococcus pneumoniae and Haemophilus influenzae during the 2-year study. Of the 16 colonization episodes, 12 (75%) were of primary endogenous pathogenesis, ie, caused by microorganisms present in the oropharynx at the time of tracheostomy. Community microorganisms including S pneumoniae, H influenzae, Moraxella (Branhamella) catarrhalis, and Staphylococcus aureus were the predominating bacteria. Three patients acquired nosocomial bacteria Pseudomonas aeruginosa and Hafnia alvei in the oropharynx, subsequently followed by secondary colonization of the lower airways. There was one failure of the prophylaxis: one patient (4%) had exogenous colonization with Pseudomonas pickettii. CONCLUSION: Topical antibiotics applied to the tracheostoma were found to be effective in reducing the exogenous route of colonization of the lower respiratory tract, compared with clinical experience and the literature. This promising technique requires further evaluation in randomized trials. PMID- 10669699 TI - beta-agonistic bronchodilators: comparison of dose/response in working rat hearts. AB - STUDY OBJECTIVES: Different beta-agonists are compared with regard to their cardiodepressive side effects. DESIGN: The metaphenolic bronchodilators reproterol, salbutamol, fenoterol, and terbutaline were introduced at a dosage of 0.0005 micromol to a maximum of 10 micromol per gram of heart tissue into the isolated working rat heart under hypoxic conditions, and the response was observed during subsequent reoxygenation. As an index of external heart work, aortic flow was measured. Heart rate, coronary flow, and developed pressure were recorded. At the end of heart perfusion, mitochondria were isolated and analyzed for adenosine triphosphatase activity, adenosine triphosphate (ATP) synthesis, and membrane fluidity. Moreover, intact mitochondria and lipid peroxidation were investigated using a model system. MEASUREMENTS AND RESULTS: Compared to controls, reproterol gave the most favorable results, with an increase of 25 to 30% of aortic flow during reoxygenation at a concentration of 10 micromol/g heart tissue. In contrast, both fenoterol and salbutamol at a concentration of 1 micromol/g heart tissue decreased aortic flow during reoxygenation, whereas terbutaline had a negative influence on aortic flow at 0.01 to 0.1 micromol/g heart tissue. Mitochondria of these hearts were isolated at the end of the experiment. Mitochondrial ATP synthesis was increased above controls at nearly all concentrations of reproterol. ATP synthesis was decreased at 1 micromol and 10 micromol fenoterol. As little as 0.0005 micromol terbutaline decreased ATP synthesis by 50%. In intact mitochondria, adenosine diphosphate (ADP) to oxygen ratios were found to be increased with terbutaline and fenoterol, indicating ADP consumption by myokinase activation. Lipid peroxidation was increased in a model system between concentrations of 0.002 micromol/mg and 0.04 micromol/mg phosphatidylcholine by fenoterol and terbutaline, whereas a decrease was noted with reproterol. Membrane fluidity was found increased after addition of reproterol, which supports the evidence of efficient ATP synthesis by this compound. CONCLUSIONS: Cardiodepressive side effects and greater toxicity of fenoterol and terbutaline were found under the conditions of our experiment. Salbutamol and, in particular, reproterol appear much better tolerated. In addition to partial beta-adrenergic agonism, reproterol may exert an inhibitory influence on adenosine receptor sites and phosphodiesterase, which could result in membrane stabilization by saving cyclic adenosine monophosphate or ATP. PMID- 10669700 TI - The drug-resistant pneumococcus: clinical relevance, therapy, and prevention. AB - Streptococcus pneumoniae has been known for > 100 years as the most important bacterial pathogen of the respiratory tract in adults and children. In recent years, the pneumococcus has begun to exhibit increasing resistance to antimicrobial agents. Because of the huge number of infections caused by this organism, the development of resistance has changed the approach to many infectious disease problems, particularly with regard to empiric antibiotic therapy and prophylaxis. In our review of the antibiotic-resistant pneumococcus, we review the microbiologic basis for resistance, risk factors for and clinical relevance of infection by a resistant organism, and infection control measures. PMID- 10669701 TI - Inhaled corticosteroids for asthma therapy: patient compliance, devices, and inhalation technique. AB - BACKGROUND: Patient compliance, inhalation devices, and inhalation techniques influence the effectiveness of inhaled medications. METHODS: This article presents the results of a systematic literature review of studies measuring compliance with inhaled corticosteroids, measuring inhalation technique with different inhalation devices, and estimating the proportion of inhaled drug that is deposited in the lung. RESULTS: Overall, patients took the recommended doses of inhaled medication on 20 to 73% of days. Frequency of efficient inhalation technique ranged from 46 to 59% of patients. Education programs have been shown to improve compliance and inhalation techniques. The lung deposition achieved with different inhalers depends on particle size as well as inhaler technique. CONCLUSION: This review demonstrates that multiple factors may come between a prescription of an inhaled corticosteroid and the arrival of that medicine at its target organ, the lung. PMID- 10669702 TI - Neuraxial blockade and hematoma in cardiac surgery: estimating the risk of a rare adverse event that has not (yet) occurred. PMID- 10669703 TI - Long-term value of exercise testing after acute myocardial infarction: influence of thrombolytic therapy. AB - OBJECTIVES: To evaluate the long-term predictive value of exercise testing performed early after acute myocardial infarction (AMI) in patients receiving thrombolytic therapy. DESIGN: Nonblinded prospective follow-up study. SETTING: Cardiac rehabilitation unit in a 900-bed university hospital. SUBJECTS: Four hundred forty-three patients allowed to perform exercise testing 3 weeks after AMI were followed for a median of 75 months; 183 received IV thrombolysis and 263 did not. RESULTS: Cardiac death hazard ratios were significantly increased in the presence of reduced physical working capacity on exertion, left ventricular dysfunction, and > or = 1-mm (but < 2-mm) ST-segment depression on exertion. In the group receiving thrombolytic therapy, no patient with > or = 2-mm ST-segment depression on exercise died; this group was characterized by a high rate of revascularization, whereas the group with > or = 1-mm but < 2-mm ST-segment depression was not. No parameter related to clinical or exercise testing predicted recurrent infarction in the group receiving thrombolytic therapy. Among patients not receiving thrombolysis, cardiac death was significantly related to > or = 2-mm ST-segment depression on exertion, to reduced physical working capacity, and to the lack of revascularization during follow-up. CONCLUSION: Exercise test-derived parameters have variable value in predicting long-term survival of patients performing exercise test after AMI depending on the following: (1) whether thrombolytic therapy was given or not; (2) the degree of ST-segment depression during exercise testing; and (3) the rate of revascularization. PMID- 10669704 TI - Clinical conference on management dilemmas: progressive infiltrates and respiratory failure. PMID- 10669705 TI - A prospective study of fever and bacteremia after flexible fiberoptic bronchoscopy in children. AB - STUDY OBJECTIVES: To assess the incidence of fever and bacteremia after fiberoptic bronchoscopy in immunocompetent children. DESIGN: Prospective study. PATIENTS: Immunocompetent children undergoing fiberoptic bronchoscopy between January 1997 and June 1998. MEASUREMENTS AND RESULTS: Ninety-one children were included in the study. Forty-four children (48%) developed fever within 24 h following bronchoscopy. Bacteremia was not detected in any of the cases at the time of the fever. Children who developed fever were younger than those who remained afebrile (mean age, 2.4 +/- 3.6 years vs 4.2 +/- 3.7 years; p = 0.025). In the fever group, 66% of the bronchoscopies were considered abnormal, compared to 45% in the nonfever group (p = 0.04). Of the fever group, 40.5% of BAL fluid cultures had significant bacterial growth, significantly higher compared to the nonfever group (13.2%; p = 0.006). Of the 80 patients in whom BAL was performed, fever occurred in 52.5% compared to only 18.2% in those who did not have BAL (p = 0.03). BAL fluid content of cell count, lipid-laden macrophages, and interleukin 8 were not significantly different in both groups. In a logistic regression analysis, the significant predictors for developing fever were positive bacterial culture (relative risk, 5.1; 95% confidence interval, 1.6 to 16.4; p = 0.007) and abnormal bronchoscopic findings (relative risk, 3.1, 95% confidence interval, 1.2 to 8.3; p = 0.02). When age < 2 years was included in the model, this factor became highly significant (relative risk, 5.01; 95% confidence interval, 1.83 to 13.75; p < 0.002). CONCLUSIONS: Fever following fiberoptic bronchoscopy is a common event in immunocompetent children and is not associated with bacteremia. Risks to develop this complication are age < 2 years, positive bacterial cultures in BAL fluid, and abnormal bronchoscopic findings. PMID- 10669706 TI - The invaluable pressure-volume curve. AB - We present a case in which the pressure-volume (P-V) curve proved invaluable in the diagnostic workup of a patient. The patient was a 43-year-old man who presented with progressive dyspnea on exertion, restrictive spirometry, exercise desaturation, and an unremarkable CT scan. Because of the unexpected finding of an unremarkable CT scan, we wanted more data assuring the presence of an indication for lung biopsy. Detailed pulmonary function tests, including a P-V curve, were administered. The P-V curve was abnormal, thus prompting a biopsy, which revealed hypersensitivity pneumonitis. In this report, we discuss the use of P-V curves and the clinical presentation of hypersensitivity pneumonitis. PMID- 10669707 TI - A 69-year-Old woman with CREST syndrome, dyspnea, and a mosaic CT attenuation pattern. PMID- 10669708 TI - Persistant right lower lobe consolidation. PMID- 10669709 TI - Complete remission of pulmonary spindle cell carcinoma after treatment with oral germanium sesquioxide. AB - Spindle cell carcinoma (SCC) is a rare form of lung cancer representing 0.2 to 0.3% of all primary pulmonary malignancies. Even with combined surgery, chemotherapy, and radiation therapy, these tumors are associated with a poor prognosis and only 10% of patients survive 2 years after diagnosis. We describe a patient with an unresectable SCC who, following no response to conventional treatment with combined modality therapy, chose to medicate herself with daily doses of germanium obtained in a health food store. She noted prompt symptomatic improvement and remains clinically and radiographically free of disease 42 months after starting her alternative therapy. PMID- 10669710 TI - Asthma and Cushing's syndrome. AB - A female patient was treated with high-dose inhaled fluticasone propionate for her asthma. Over 2 years, she developed features of Cushing's syndrome with proximal myopathy, osteopenia, hypertension, depressive psychosis, and cushingoid appearance. She had biochemical evidence of marked adrenal suppression with a 9:00 AM serum cortisol of 20 nmol/L that returned to normal (315 mol/L) after her therapy was changed to budenoside, 0.8 mg/d. Her appearance, mental state, and myopathy also improved with no loss of asthma control. This case illustrates the potential for developing clinically relevant adverse effects of inhaled corticosteroids when given at licensed doses. PMID- 10669711 TI - Acute massive pulmonary embolism in a Jehovah's witness: successful treatment with catheter thrombectomy. AB - A 71-year-old woman presented with an acute, massive pulmonary embolism. As a Jehovah's Witness, she was not willing to accept thrombolysis because of the potential risk of bleeding requiring blood transfusion. The patient was successfully treated with catheter thrombectomy, using rheolytic and fragmentation devices. (CHEST 2000; 117:594-597) PMID- 10669712 TI - Carbamazepine-induced systemic lupus erythematosus presenting as cardiac tamponade. AB - Here we report the case of a patient who presented with acute cardiac tamponade due to drug-induced systemic lupus erythematosus (SLE). The patient had been treated for a seizure disorder with carbamazepine, a drug that has previously been demonstrated to cause SLE-like syndromes. Further serologic analysis demonstrated the likelihood of drug-induced SLE in this patient, with the rare presentation of cardiac tamponade. PMID- 10669713 TI - Pedal (99m)Tc-sulfur colloid lymphoscintigraphy in primary isolated chylopericardium. AB - Primary isolated chylopericardium is a rare disorder in which chylous fluid accumulates in the pericardial space. In this case report of a 61-year-old man with chylopericardium, pedal (99m)Tc-sulfur colloid (SC) lymphoscintigraphy was performed after emergent pericardiocentesis, and when there was a recurrent massive pericardial effusion. The results showed that (99m)Tc-SC lymphoscintigraphy can clearly reveal the lymphodynamics in patients with primary isolated chylopericardium. This noninvasive investigation is valuable and can be easily performed either before or after pericardiocentesis. PMID- 10669714 TI - A large false aneurysm of the right ventricle within a giant epicardial lipoma. AB - Lipomas, which account for approximately 10% of all neoplasms of the heart, may be detected in asymptomatic patients by chance during echocardiography, CT scan, or MRI scan. Occasionally, lipomas are complicated by arrhythmias. We describe a patient who presented with severe cardiomegaly and paroxysmal supraventricular tachycardia. An MRI scan showed a large intrapericardial lipoma with two large cavities inside communicating with each other and with the right ventricular chamber through a defect of the right ventricular wall. The mass was partially removed, and the right ventricle was patched. Surgery combined with antiarrhythmic therapy resulted in a good short-term result. PMID- 10669715 TI - Paraneoplastic pemphigus associated with bronchiolitis obliterans. AB - Paraneoplastic pemphigus (PNP) is an autoantibody-mediated mucocutaneous blistering disease associated with underlying neoplasms. Autoantibodies of PNP bind to the plakin family of cytoplasmic proteins and desmogleins of cell-surface target antigens. We describe a 36-year-old female patient with PNP who had non Hodgkin's lymphoma, and who developed bronchiolitis obliterans and died of respiratory failure. Autopsy findings confirmed luminal narrowing of bronchioles by scarring, which is a histopathologic features of bronchiolitis obliterans. After the onset of respiratory failure, the reaction of autoantibodies against the plakins detected by immunoprecipitation at the onset of PNP disappeared with negative immunofluorescence within the bronchial epithelium. It is thought that autoantibodies against some of these antigens play a role in causing acute inflammation of the respiratory epithelium. In treating PNP, the possibility of the patient developing the lethal complication bronchiolitis obliterans should be kept in mind. Furthermore, prevention of the initial autoantibody-mediated injury to the respiratory epithelium should be an important treatment goal. PMID- 10669716 TI - Factors contributing to pneumothorax after thoracentesis. PMID- 10669717 TI - Longitudinal data on positive tuberculin skin tests from three US states. PMID- 10669718 TI - Salmeterol and tolerance. PMID- 10669719 TI - Use of fluticasone in asthma. PMID- 10669720 TI - Confident diagnosis of solitary fibrous tumor of the pleura using cutting-needle biopsy. PMID- 10669721 TI - Fatal pulmonary hemorrhage during high-dose valproate monotherapy. PMID- 10669722 TI - The mechanism of hypoxemia in liver disease with pulmonary hypertension. PMID- 10669723 TI - Community-acquired chlamydia pneumoniae pneumonia. PMID- 10669725 TI - TATA-binding protein mutants that increase transcription from enhancerless and repressed promoters in vivo. AB - Using a genetic screen, we isolated three TATA-binding protein (TBP) mutants that increase transcription from promoters that are repressed by the Cyc8-Tup1 or Sin3 Rpd3 corepressors or that lack an enhancer element, but not from an equivalently weak promoter with a mutated TATA element. Increased transcription is observed when the TBP mutants are expressed at low levels in the presence of wild-type TBP. These TBP mutants are unable to support cell viability, and they are toxic in strains lacking Rpd3 histone deacetylase or when expressed at higher levels. Although these mutants do not detectably bind TATA elements in vitro, genetic and chromatin immunoprecipitation experiments indicate that they act directly at promoters and do not increase transcription by titration of a negative regulatory factor(s). The TBP mutants are mildly defective for associating with promoters responding to moderate or strong activators; in addition, they are severely defective for RNA polymerase (Pol) III but not Pol I transcription. These results suggest that, with respect to Pol II transcription, the TBP mutants specifically increase expression from core promoters. Biochemical analysis indicates that the TBP mutants are unaffected for TFIID complex formation, dimerization, and interactions with either the general negative regulator NC2 or the N-terminal inhibitory domain of TAF130. We speculate that these TBP mutants have an unusual structure that allows them to preferentially access TATA elements in chromatin templates. These TBP mutants define a criterion by which promoters repressed by Cyc8-Tup1 or Sin3-Rpd3 resemble enhancerless, but not TATA-defective, promoters; hence, they support the idea that these corepressors inhibit the function of activator proteins rather than the Pol II machinery. PMID- 10669724 TI - Regulatory and signaling properties of the Vav family. PMID- 10669726 TI - Insulin-like growth factor I-induced degradation of insulin receptor substrate 1 is mediated by the 26S proteasome and blocked by phosphatidylinositol 3'-kinase inhibition. AB - Insulin receptor substrate 1 (IRS-1) is a critical adapter protein involved in both insulin and insulin-like growth factor (IGF) signaling. Due to the fact that alteration of IRS-1 levels can affect the sensitivity and response to both insulin and IGF-I, we examined the ability of each of these ligands to affect IRS 1 expression. IGF-I (10 nM) stimulation of MCF-7 breast cancer cells caused a transient tyrosine phosphorylation of IRS-1 that was maximal at 15 min and decreased thereafter. The decrease in tyrosine phosphorylation of IRS-1 was paralleled by an apparent decrease in IRS-1 levels. The IGF-mediated decrease in IRS-1 expression was posttranscriptional and due to a decrease in the half-life of the IRS-1 protein. Insulin (10 nM) caused tyrosine phosphorylation of IRS-1 but not degradation, whereas high concentrations of insulin (10 microM) resulted in degradation of IRS-1. IGF-I (10 nM) stimulation resulted in transient IRS-1 phosphorylation and extracellular signal-related kinase (ERK) activation. In contrast, insulin (10 nM) caused sustained IRS-1 phosphorylation and ERK activation. Inhibition of 26S proteasome activity by the use of lactacystin or MG132 completely blocked IGF-mediated degradation of IRS-1. Furthermore, coimmunoprecipitation experiments showed an association between ubiquitin and IRS 1 that was increased by treatment of cells with IGF-I. Finally, IGF-mediated degradation of IRS-1 was blocked by inhibition of phosphatidylinositol 3'-kinase activity but was not affected by inhibition of ERK, suggesting that this may represent a direct negative-feedback mechanism resulting from downstream IRS-1 signaling. We conclude that IGF-I can cause ligand-mediated degradation of IRS-1 via the ubiquitin-mediated 26S proteasome and a phosphatidylinositol 3'-kinase dependent mechanism and that control of degradation may have profound effects on downstream activation of signaling pathways. PMID- 10669727 TI - Distinct roles for Galpha(i)2 and Gbetagamma in signaling to DNA synthesis and Galpha(i)3 in cellular transformation by dopamine D2S receptor activation in BALB/c 3T3 cells. AB - Control of cell proliferation depends on intracellular mediators that determine the cellular response to external cues. In neuroendocrine cells, the dopamine D2 receptor short form (D2S receptor) inhibits cell proliferation, whereas in mesenchymal cells the same receptor enhances cell proliferation. Nontransformed BALB/c 3T3 fibroblast cells were stably transfected with the D2S receptor cDNA to study the G proteins that direct D2S signaling to stimulate cell proliferation. Pertussis toxin inactivates G(i) and G(o) proteins and blocks signaling of the D2S receptor in these cells. D2S receptor signaling was reconstituted by individually transfecting pertussis toxin-resistant Galpha(i/o) subunit mutants and measuring D2-induced responses in pertussis toxin-treated cells. This approach identified Galpha(i)2 and Galpha(i)3 as mediators of the D2S receptor mediated inhibition of forskolin-stimulated adenylyl cyclase activity; Galpha(i)2 mediated D2S-induced stimulation of p42 and p44 mitogen-activated kinase (MAPK) and DNA synthesis, whereas Galpha(i)3 was required for formation of transformed foci. Transfection of toxin-resistant Galpha(i)1 cDNA induced abnormal cell growth independent of D2S receptor activation, while Galpha(o) inhibited dopamine induced transformation. The role of Gbetagamma subunits was assessed by ectopic expression of the carboxyl-terminal domain of G protein receptor kinase to selectively antagonize Gbetagamma activity. Mobilization of Gbetagamma subunits was required for D2S-induced calcium mobilization, MAPK activation, and DNA synthesis. These findings reveal a remarkable and distinct G protein specificity for D2S receptor-mediated signaling to initiate DNA synthesis (Galpha(i)2 and Gbetagamma) and oncogenic transformation (Galpha(i)3), and they indicate that acute activation of MAPK correlates with enhanced DNA synthesis but not with transformation. PMID- 10669728 TI - Caveolin 1-mediated regulation of receptor tyrosine kinase-associated phosphatidylinositol 3-kinase activity by ceramide. AB - Previous studies have indicated that proapoptotic stresses downregulate the phosphatidylinositol 3-kinase [PI(3)K]/Akt survival pathway via the activation of acid-sphingomyelinase (A-SMase) and ceramide production. Ceramide induces apoptosis and inhibits PI(3)K activity without altering expression, association, or phosphorylation of receptors, adapter proteins, or PI(3)K subunits. PI(3)K inhibition by ceramide is associated with recruitment of caveolin 1 to PI(3)K associated receptor complexes within lipid raft microdomains. Overexpression of caveolin 1 alone is sufficient to alter PI(3)K activity and sensitizes fibroblasts to ceramide-induced cell death. Most importantly, antisense expression of caveolin 1 dramatically reduces ceramide-induced PI(3)K deregulation and results in a loss-of-function stress response similar to that in A-SMase-deficient cells. Stress-induced recruitment of caveolin 1 to receptor complexes was found to be dependent on A-SMase since cell lines deficient in A SMase did not exhibit caveolin 1 association with PI(3)K receptor complexes. Thus, a genetic link between A-SMase activation and caveolin 1-induced inhibition of PI(3)K activity exists. These results led us to propose that stress-induced changes in raft microdomains lead to altered receptor tyrosine kinase signal transduction through the modulation of caveolin 1 by ceramide. PMID- 10669729 TI - Complex protein interactions within the human polyadenylation machinery identify a novel component. AB - Polyadenylation of mRNA precursors is a two-step reaction requiring multiple protein factors. Cleavage stimulation factor (CstF) is a heterotrimer necessary for the first step, endonucleolytic cleavage, and it plays an important role in determining the efficiency of polyadenylation. Although a considerable amount is known about the RNA binding properties of CstF, the protein-protein interactions required for its assembly and function are poorly understood. We therefore first identified regions of the CstF subunits, CstF-77, CstF-64, and CstF-50, required for interaction with each other. Unexpectedly, small regions of two of the subunits participate in multiple interactions. In CstF-77, a proline-rich domain is necessary not only for binding both other subunits but also for self association, an interaction consistent with genetic studies in Drosophila. In CstF-64, a small region, highly conserved in metazoa, is responsible for interactions with two proteins, CstF-77 and symplekin, a nuclear protein of previously unknown function. Intriguingly, symplekin has significant similarity to a yeast protein, PTA1, that is a component of the yeast polyadenylation machinery. We show that multiple factors, including CstF, cleavage polyadenylation specificity factor, and symplekin, can be isolated from cells as part of a large complex. These and other data suggest that symplekin may function in assembly of the polyadenylation machinery. PMID- 10669730 TI - Molecular mechanism for the Shp-2 tyrosine phosphatase function in promoting growth factor stimulation of Erk activity. AB - We have previously shown that activation of extracellular signal-regulated kinase (Erk) by epidermal growth factor (EGF) treatment was significantly decreased in mouse fibroblast cells expressing a mutant Shp-2 molecule lacking 65 amino acids in the SH2-N domain, Shp-2(Delta46-110). To address the molecular mechanism for the positive role of Shp-2 in mediating Erk induction, we evaluated the activation of signaling components upstream of Erk in Shp-2 mutant cells. EGF stimulated Ras, Raf, and Mek activation was significantly attenuated in Shp-2 mutant cells, suggesting that Shp-2 acts to promote Ras activation or to suppress the down-regulation of activated Ras. Biochemical analyses indicate that upon EGF stimulation, Shp-2 is recruited into a multiprotein complex assembled on the Gab1 docking molecule and that Shp-2 seems to exert its biological function by specifically dephosphorylating an unidentified molecule of 90 kDa in the complex. The mutant Shp-2(Delta46-110) molecule failed to participate in the Gab1 organized complex for dephosphorylation of p90, correlating with a defective activation of the Ras-Raf-Mek-Erk cascade in EGF-treated Shp-2 mutant cells. Evidence is also presented that Shp-2 does not appear to modulate the signal relay from EGF receptor to Ras through the Shc, Grb2, and Sos proteins. These results begin to elucidate the mechanism of Shp-2 function downstream of a receptor tyrosine kinase to promote the activation of the Ras-Erk pathway, with potential therapeutic applications in cancer treatment. PMID- 10669731 TI - Nck-interacting Ste20 kinase couples Eph receptors to c-Jun N-terminal kinase and integrin activation. AB - The mammalian Ste20 kinase Nck-interacting kinase (NIK) specifically activates the c-Jun amino-terminal kinase (JNK) mitogen-activated protein kinase module. NIK also binds the SH3 domains of the SH2/SH3 adapter protein Nck. To determine whether Nck functions as an adapter to couple NIK to a receptor tyrosine kinase signaling pathway, we determined whether NIK is activated by Eph receptors (EphR). EphRs constitute the largest family of receptor tyrosine kinases (RTK), and members of this family play important roles in patterning of the nervous and vascular systems. In this report, we show that NIK kinase activity is specifically increased in cells stimulated by two EphRs, EphB1 and EphB2. EphB1 kinase activity and phosphorylation of a juxtamembrane tyrosine (Y594), conserved in all Eph receptors, are both critical for NIK activation by EphB1. Although pY594 in the EphB1R has previously been shown to bind the SH2 domain of Nck, we found that stimulation of EphB1 and EphB2 led predominantly to a complex between NIK/Nck, p62(dok), RasGAP, and an unidentified 145-kDa tyrosine-phosphorylated protein. Tyrosine-phosphorylated p62(dok) most probably binds directly to the SH2 domain of Nck and RasGAP and indirectly to NIK bound to the SH3 domain of Nck. We found that NIK activation is also critical for coupling EphB1R to biological responses that include the activation of integrins and JNK by EphB1. Taken together, these findings support a model in which the recruitment of the Ste20 kinase NIK to phosphotyrosine-containing proteins by Nck is an important proximal step in the signaling cascade downstream of EphRs. PMID- 10669732 TI - Recruitment of CREB binding protein is sufficient for CREB-mediated gene activation. AB - Phosphorylation of the transcription factor CREB leads to the recruitment of the coactivator, CREB binding protein (CBP). Recent studies have suggested that CBP recruitment is not sufficient for CREB function, however. We have identified a conserved protein-protein interaction motif within the CBP-binding domains of CREB and another transcription factor, SREBP (sterol-responsive element binding protein). In contrast to CREB, SREBP interacts with CBP in the absence of phosphorylation. We have exploited the conservation of this interaction motif to test whether CBP recruitment to CREB is sufficient for transcriptional activation. Substitution of six nonconserved amino acids from SREBP into the activation domain of CREB confers high-affinity, phosphorylation-independent CBP binding. The mutated CREB molecule, CREB(DIEDML), activates transcription in F9 teratocarcinoma and PC12 cells even in the absence of protein kinase A (PKA). Addition of exogenous CBP augments the level of transcription mediated by CREB(DIEDML), and adenovirus 12S E1A blocks transcription, implicating CBP in the activation process. Thus, recruitment of CBP to CREB is sufficient for transcriptional activation. Addition of PKA stimulates transcription induced by CREB(DIEDML) further, suggesting that a phosphorylation event downstream from CBP recruitment augments CREB signaling. PMID- 10669733 TI - Timing of developmentally programmed excision and circularization of Paramecium internal eliminated sequences. AB - Paramecium internal eliminated sequences (IESs) are short AT-rich DNA elements that are precisely eliminated from the germ line genome during development of the somatic macronucleus. They are flanked by one 5'-TA-3' dinucleotide on each side, a single copy of which remains at the donor site after excision. The timing of their excision was examined in synchronized conjugating cells by quantitative PCR. Significant amplification of the germ line genome was observed prior to IES excision, which starts 12 to 14 h after initiation of conjugation and extends over a 2- to 4-h period. Following excision, two IESs were shown to form extrachromosomal circles that can be readily detected on Southern blots of genomic DNA from cells undergoing macronuclear development. On these circular molecules, covalently joined IES ends are separated by one copy of the flanking 5'-TA-3' repeat. The similar structures of the junctions formed on the excised and donor molecules point to a central role for this dinucleotide in IES excision. PMID- 10669734 TI - Terminally differentiated human neurons repair transcribed genes but display attenuated global DNA repair and modulation of repair gene expression. AB - Repair of UV-induced DNA lesions in terminally differentiated human hNT neurons was compared to that in their repair-proficient precursor NT2 cells. Global genome repair of (6-4)pyrimidine-pyrimidone photoproducts was significantly slower in hNT neurons than in the precursor cells, and repair of cyclobutane pyrimidine dimers (CPDs) was not detected in the hNT neurons. This deficiency in global genome repair did not appear to be due to denser chromatin structure in hNT neurons. By contrast, CPDs were removed efficiently from both strands of transcribed genes in hNT neurons, with the nontranscribed strand being repaired unexpectedly well. Correlated with these changes in repair during neuronal differentiation were modifications in the expression of several repair genes, in particular an up-regulation of the two structure-specific nucleases XPG and XPF/ERCC1. These results have implications for neuronal dysfunction and aging. PMID- 10669735 TI - Nuclear import of IkappaBalpha is accomplished by a ran-independent transport pathway. AB - The inhibitor of kappa B alpha (IkappaBalpha) protein is able to shuttle between the cytoplasm and the nucleus. We have utilized a combination of in vivo and in vitro approaches to provide mechanistic insight into nucleocytoplasmic shuttling by IkappaBalpha. IkappaBalpha contains multiple functional domains that contribute to shuttling of IkappaBalpha between the cytoplasm and the nucleus. Nuclear import of IkappaBalpha is mediated by the central ankyrin repeat domain. Similar to previously described nuclear import pathways, nuclear import of IkappaBalpha is temperature and ATP dependent and is blocked by a dominant negative mutant of importin beta. However, in contrast to classical nuclear import pathways, nuclear import of IkappaBalpha is independent of soluble cytosolic factors and is not blocked by the dominant-negative RanQ69L protein. Nuclear export of IkappaBalpha is mediated by an N-terminal nuclear export sequence. Nuclear export of IkappaBalpha requires the CRM1 nuclear export receptor and is blocked by the dominant-negative RanQ69L protein. Our results are consistent with a model in which nuclear import of IkappaBalpha is mediated through direct interactions with components of the nuclear pore complex, while nuclear export of IkappaBalpha is mediated via a CRM1-dependent pathway. PMID- 10669736 TI - Transient expression of cellular polypyrimidine-tract binding protein stimulates cap-independent translation directed by both picornaviral and flaviviral internal ribosome entry sites In vivo. AB - The regulation of cap-independent translation directed by the internal ribosome entry sites (IRESs) present in some viral and cellular RNAs is poorly understood. Polypyrimidine-tract binding protein (PTB) binds specifically to several viral IRESs. IRES-directed translation may be reduced in cell-free systems that are depleted of PTB and restored by reconstitution of lysates with recombinant PTB. However, there are no data concerning the effects of PTB on IRES-directed translation in vivo. We transfected cells with plasmids expressing dicistronic transcripts in which the upstream cistron encoded PTB or PTB deletion mutants (including a null mutant lacking amino acid residues 87 to 531). The downstream cistron encoded a reporter protein (chloramphenicol acetyltransferase [CAT]) under translational control of the poliovirus IRES which was placed within the intercistronic space. In transfected BS-C-1 cells, transcripts expressing wild type PTB produced 12-fold more reporter protein than similar transcripts encoding the PTB null mutant. There was a 2.4-fold difference in CAT produced from these transcripts in HeLa cells, which contain a greater natural abundance of PTB. PTB similarly stimulated CAT production from transcripts containing the IRES of hepatitis A virus or hepatitis C virus in BS-C-1 cells and Huh-7 cells (37- to 44 fold increase and 5 to 5.3-fold increase, respectively). Since PTB had no quantitative or qualitative effect on transcription from these plasmids, we conclude that PTB stimulates translation of representative picornaviral and flaviviral RNAs in vivo. This is likely to reflect the stabilization of higher ordered RNA structures within the IRES and was not observed with PTB mutants lacking RNA recognition motifs located in the C-terminal third of the molecule. PMID- 10669737 TI - The phosphorylation status of a cyclic AMP-responsive activator is modulated via a chromatin-dependent mechanism. AB - Cyclic AMP (cAMP) stimulates the expression of numerous genes via the protein kinase A (PKA)-mediated phosphorylation of CREB at Ser133. Ser133 phosphorylation, in turn, promotes recruitment of the coactivator CREB binding protein and its paralog p300, histone acetyltransferases (HATs) that have been proposed to mediate target gene activation, in part, by destabilizing promoter bound nucleosomes and thereby allowing assembly of the transcriptional apparatus. Here we show that although histone deacetylase (HDAC) inhibitors potentiate target gene activation via cAMP, they do not stimulate transcription over the early burst phase, during which CREB phosphorylation and CBP/p300 recruitment are maximal. Rather, HDAC inhibitors augment CREB activity during the late attenuation phase by prolonging CREB phosphorylation on chromosomal but, remarkably, not on extrachromosomal templates. In reconstitution studies, assembly of periodic nucleosomal arrays on a cAMP-responsive promoter template potently inhibited CREB phosphorylation by PKA, and acetylation of these template bound nucleosomes by p300 partially rescued CREB phosphorylation by PKA. Our results suggest a novel regulatory mechanism by which cellular HATs and HDACs modulate the phosphorylation status of nuclear activators in response to cellular signals. PMID- 10669738 TI - The putative pore-forming domain of Bax regulates mitochondrial localization and interaction with Bcl-X(L). AB - Bax is a proapoptotic member of the Bcl-2 family of proteins which localizes to and uses mitochondria as its major site of action. Bax normally resides in the cytoplasm and translocates to mitochondria in response to apoptotic stimuli, and it promotes apoptosis in two ways: (i) by disrupting mitochondrial membrane barrier function by formation of ion-permeable pores in mitochondrial membranes and (ii) by binding to antiapoptotic Bcl-2 family proteins via its BH3 domain and inhibiting their functions. A hairpin pair of amphipathic alpha-helices (alpha5 alpha6) in Bax has been predicted to participate in membrane insertion and pore formation by Bax. We mutagenized several charged residues in the alpha5-alpha6 domain of Bax, changing them to alanine. These substitution mutants of Bax constitutively localized to mitochondria and displayed a gain-of-function phenotype when expressed in mammalian cells. Furthermore, substitution of 8 out of 10 charged residues in the alpha5-alpha6 domain of Bax resulted in a loss of cytotoxicity in yeast but a gain-of-function phenotype in mammalian cells. The enhanced function of this Bax mutant was correlated with increased binding to Bcl X(L), through a BH3-independent mechanism. These observations reveal new functions for the alpha5-alpha6 hairpin loop of Bax: (i) regulation of mitochondrial targeting and (ii) modulation of binding to antiapoptotic Bcl-2 proteins. PMID- 10669739 TI - Cubitus interruptus requires Drosophila CREB-binding protein to activate wingless expression in the Drosophila embryo. AB - CREB-binding protein (CBP) serves as a transcriptional coactivator in multiple signal transduction pathways. The Drosophila homologue of CBP, dCBP, interacts with the transcription factors Cubitus interruptus (CI), MAD, and Dorsal (DL) and functions as a coactivator in several signaling pathways during Drosophila development, including the hedgehog (hh), decapentaplegic (dpp), and Toll pathways. Although dCBP is required for the expression of the hh target genes, wingless (wg) and patched (ptc) in vivo, and potentiates ci-mediated transcriptional activation in vitro, it is not known that ci absolutely requires dCBP for its activity. We used a yeast genetic screen to identify several ci point mutations that disrupt CI-dCBP interactions. These mutant proteins are unable to transactivate a reporter gene regulated by ci binding sites and have a lower dCBP-stimulated activity than wild-type CI. When expressed exogenously in embryos, the CI point mutants cannot activate endogenous wg expression. Furthermore, a CI mutant protein that lacks the entire dCBP interaction domain functions as a negative competitor for wild-type CI activity, and the expression of dCBP antisense RNAs can suppress CI transactivation in Kc cells. Taken together, our data suggest that dCBP function is necessary for ci-mediated transactivation of wg during Drosophila embryogenesis. PMID- 10669741 TI - Two novel Drosophila TAF(II)s have homology with human TAF(II)30 and are differentially regulated during development. AB - TFIID is a multiprotein complex composed of the TATA binding protein (TBP) and TBP-associated factors (TAF(II)s). The binding of TFIID to the promoter is the first step of RNA polymerase II preinitiation complex assembly on protein-coding genes. Yeast (y) and human (h) TFIID complexes contain 10 to 13 TAF(II)s. Biochemical studies suggested that the Drosophila (d) TFIID complexes contain only eight TAF(II)s, leaving a number of yeast and human TAF(II)s (e.g., hTAF(II)55, hTAF(II)30, and hTAF(II)18) without known Drosophila homologues. We demonstrate that Drosophila has not one but two hTAF(II)30 homologues, dTAF(II)16 and dTAF(II)24, which are encoded by two adjacent genes. These two genes are localized in a head-to-head orientation, and their 5' extremities overlap. We show that these novel dTAF(II)s are expressed and that they are both associated with TBP and other bona fide dTAF(II)s in dTFIID complexes. dTAF(II)24, but not dTAF(II)16, was also found to be associated with the histone acetyltransferase (HAT) dGCN5. Thus, dTAF(II)16 and dTAF(II)24 are functional homologues of hTAF(II)30, and this is the first demonstration that a TAF(II)-GCN5-HAT complex exists in Drosophila. The two dTAF(II)s are differentially expressed during embryogenesis and can be detected in both nuclei and cytoplasm of the cells. These results together indicate that dTAF(II)16 and dTAF(II)24 may have similar but not identical functions. PMID- 10669740 TI - Akt suppresses apoptosis by stimulating the transactivation potential of the RelA/p65 subunit of NF-kappaB. AB - It is well established that cell survival signals stimulated by growth factors, cytokines, and oncoproteins are initiated by phosphoinositide 3-kinase (PI3K)- and Akt-dependent signal transduction pathways. Oncogenic Ras, an upstream activator of Akt, requires NF-kappaB to initiate transformation, at least partially through the ability of NF-kappaB to suppress transformation-associated apoptosis. In this study, we show that oncogenic H-Ras requires PI3K and Akt to stimulate the transcriptional activity of NF-kappaB. Activated forms of H-Ras and MEKK stimulate signals that result in nuclear translocation and DNA binding of NF kappaB as well as stimulation of the NF-kappaB transactivation potential. In contrast, activated PI3K or Akt stimulates NF-kappaB-dependent transcription by stimulating transactivation domain 1 of the p65 subunit rather than inducing NF kappaB nuclear translocation via IkappaB degradation. Inhibition of IkappaB kinase (IKK), using an IKKbeta dominant negative protein, demonstrated that activated Akt requires IKK to efficiently stimulate the transactivation domain of the p65 subunit of NF-kappaB. Inhibition of endogenous Akt activity sensitized cells to H-Ras(V12)-induced apoptosis, which was associated with a loss of NF kappaB transcriptional activity. Finally, Akt-transformed cells were shown to require NF-kappaB to suppress the ability of etoposide to induce apoptosis. Our work demonstrates that, unlike activated Ras, which can stimulate parallel pathways to activate both DNA binding and the transcriptional activity of NF kappaB, Akt stimulates NF-kappaB predominantly by upregulating of the transactivation potential of p65. PMID- 10669742 TI - CIZ, a zinc finger protein that interacts with p130(cas) and activates the expression of matrix metalloproteinases. AB - p130(cas) (Cas) is a docking protein that contains an SH3 domain and multiple tyrosine residues. p130(cas) is located at focal adhesions, is tyrosine phosphorylated in response to integrin stimulation, and is thought to transmit signals, via c-Crk and other proteins, for the remodeling of actin stress fibers and cell movement. In a search for the ligands of the SH3 domain of p130(cas) by far-Western screening, we cloned a novel protein named CIZ (for Cas-interacting zinc finger protein). CIZ consists of the following: a putative leucine zipper; a serine/threonine-rich region; a proline-rich sequence; five, six, or eight Kruppel-type C(2)H(2) zinc fingers; and the glutamine-alanine repeat. CIZ binds Cas in cells and is located in the nucleus and at focal adhesions. We showed that CIZ is a nucleocytoplasmic shuttling protein, by using the transient interspecies heterokaryon formation assay. In order to search for the targets of CIZ in nucleus, we determined the DNA binding consensus of CIZ as (G/C)AAAAA(A) by cyclic amplification and selection of targets analysis. The consensus-like sequences are found in several promoters of matrix metalloproteinases (MMPs), which are the enzymes used to degrade the extracellular matrix proteins. CIZ binds to a consensus-like sequence in the MMP-1 (collagenase) promoter. Overexpression of CIZ upregulates the transcriptions from MMP-1, MMP-3 (stromelysin), and MMP-7 (matrilysin) promoters, and this transactivation was enhanced in the presence of Cas. Furthermore, the stable overexpression of CIZ promoted the production of MMP-7 in culture medium. In summary, CIZ, a novel zinc finger protein, binds Cas, is a nucleocytoplasmic shuttling protein, and regulates the expression of MMPs. PMID- 10669743 TI - Control of human telomere length by TRF1 and TRF2. AB - Telomere length in human cells is controlled by a homeostasis mechanism that involves telomerase and the negative regulator of telomere length, TRF1 (TTAGGG repeat binding factor 1). Here we report that TRF2, a TRF1-related protein previously implicated in protection of chromosome ends, is a second negative regulator of telomere length. Overexpression of TRF2 results in the progressive shortening of telomere length, similar to the phenotype observed with TRF1. However, while induction of TRF1 could be maintained over more than 300 population doublings and resulted in stable, short telomeres, the expression of exogenous TRF2 was extinguished and the telomeres eventually regained their original length. Consistent with their role in measuring telomere length, indirect immunofluorescence indicated that both TRF1 and TRF2 bind to duplex telomeric DNA in vivo and are more abundant on telomeres with long TTAGGG repeat tracts. Neither TRF1 nor TRF2 affected the expression level of telomerase. Furthermore, the presence of TRF1 or TRF2 on a short linear telomerase substrate did not inhibit the enzymatic activity of telomerase in vitro. These findings are consistent with the recently proposed t loop model of telomere length homeostasis in which telomerase-dependent telomere elongation is blocked by sequestration of the 3' telomere terminus in TRF1- and TRF2-induced telomeric loops. PMID- 10669744 TI - Sarcospan-deficient mice maintain normal muscle function. AB - Sarcospan is an integral membrane component of the dystrophin-glycoprotein complex (DGC) found at the sarcolemma of striated and smooth muscle. The DGC plays important roles in muscle function and viability as evidenced by defects in components of the DGC, which cause muscular dystrophy. Sarcospan is unique among the components of the complex in that it contains four transmembrane domains with intracellular N- and C-terminal domains and is a member of the tetraspan superfamily of proteins. Sarcospan is tightly linked to the sarcoglycans, and together these proteins form a subcomplex within the DGC. Stable expression of sarcospan at the sarcolemma is dependent upon expression of the sarcoglycans. Here we describe the generation and analysis of mice carrying a null mutation in the Sspn gene. Surprisingly, the Sspn-deficient muscle maintains expression of other components of the DGC at the sarcolemma, and no gross histological abnormalities of muscle from the mice are observed. The Sspn-deficient muscle maintains sarcolemmal integrity as determined by serum creatine kinase and Evans blue uptake assays, and the Sspn-deficient muscle maintains normal force and power generation capabilities. These data suggest either that sarcospan is not required for normal DGC function or that the Sspn-deficient muscle is compensating for the absence of sarcospan, perhaps by utilizing another protein to carry out its function. PMID- 10669745 TI - Tyrosine phosphorylation mediates both activation and downmodulation of the biological activity of Vav. AB - Vav works as a GDP/GTP exchange factor for Rac GTPases, thereby facilitating the transition of these proteins from the inactive (GDP-bound) into the active (GTP bound) state. The stimulation of Vav exchange activity during cell signaling is mediated by tyrosine phosphorylation. To understand the roles of phosphorylation in the regulation of Vav activity, we have initiated the characterization of the residues of Vav that are phosphorylated during signal transduction. Here we show that a Y-to-F mutation in one of these residues, Y174, leads to the oncogenic activation of Vav and to the enhancement of other Vav-mediated signals such as those for cytoskeletal reorganization, JNK activation, and stimulation of the nuclear factor of activated T cells. The effect induced by the Y174F mutation is further accentuated by mutations in residue Y142 or Y160. The Y174F mutation has no effect on the exchange activity of Vav in vitro but results in higher levels of phosphorylation in vivo. Using a phosphospecific antibody, we found that Y174 is phosphorylated following stimulation of mitogenic and antigenic receptors. This phosphorylation event is conserved in Vav-2 and Vav-3, the other two members of the Vav family. These results identify a previously unknown mechanism for the oncogenic activation of Vav and suggest that the activity of this exchange factor is modulated by two antagonistic phosphorylation events, one involved in Vav activation and a second one implicated in Vav inactivation. PMID- 10669746 TI - Inhibition of IkappaB kinase and IkappaB phosphorylation by 15-deoxy-Delta(12,14) prostaglandin J(2) in activated murine macrophages. AB - Activation of the macrophage cell line RAW 264.7 with lipopolysaccharide (LPS) and gamma interferon (IFN-gamma) induces the expression of gene products involved in host defense, among them type 2 nitric oxide synthase. Treatment of cells with 15-deoxy-Delta(12,14)-prostaglandin J(2) (15dPGJ(2)) inhibited the LPS- and IFN gamma-dependent synthesis of NO, a process that was not antagonized by similar concentrations of prostaglandin J(2), prostaglandin E(2), or rosiglitazone, a peroxisomal proliferator-activated receptor gamma ligand. Incubation of activated macrophages with 15dPGJ(2) inhibited the degradation of IkappaBalpha and IkappaBbeta and increased their levels in the nuclei. NF-kappaB activity, as well as the transcription of NF-kappaB-dependent genes, such as those encoding type 2 nitric oxide synthase and cyclooxygenase 2, was impaired under these conditions. Analysis of the steps leading to IkappaB phosphorylation showed an inhibition of IkappaB kinase by 15dPGJ(2) in cells treated with LPS and IFN-gamma, resulting in an impaired phosphorylation of IkappaBalpha, at least in the serine 32 residue required for targeting and degradation of this protein. Incubation of partially purified activated IkappaB kinase with 2 microM 15dPGJ(2) reduced by 83% the phosphorylation in serine 32 of IkappaBalpha, suggesting that this prostaglandin exerts direct inhibitory effects on the activity of the IkappaB kinase complex. These results show rapid actions of 15dPGJ(2), independent of peroxisomal proliferator receptor gamma activation, in macrophages challenged with low doses of LPS and IFN-gamma. PMID- 10669747 TI - Cloning and characterization of SCHIP-1, a novel protein interacting specifically with spliced isoforms and naturally occurring mutant NF2 proteins. AB - The neurofibromatosis type 2 (NF2) protein, known as schwannomin or merlin, is a tumor suppressor involved in NF2-associated and sporadic schwannomas and meningiomas. It is closely related to the ezrin-radixin-moesin family members, implicated in linking membrane proteins to the cytoskeleton. The molecular mechanism allowing schwannomin to function as a tumor suppressor is unknown. In attempt to shed light on schwannomin function, we have identified a novel coiled coil protein, SCHIP-1, that specifically associates with schwannomin in vitro and in vivo. Within its coiled-coil region, this protein is homologous to human FEZ proteins and the related Caenorhabditis elegans gene product UNC-76. Immunofluorescent staining of transiently transfected cells shows a partial colocalization of SCHIP-1 and schwannomin, beneath the cytoplasmic membrane. Surprisingly, immunoprecipitation assays reveal that in a cellular context, association with SCHIP-1 can be observed only with some naturally occurring mutants of schwannomin, or a schwannomin spliced isoform lacking exons 2 and 3, but not with the schwannomin isoform exhibiting growth-suppressive activity. Our observations suggest that SCHIP-1 interaction with schwannomin is regulated by conformational changes in schwannomin, possibly induced by posttranslational modifications, alternative splicing, or mutations. PMID- 10669748 TI - Inhibition of c-Jun N-terminal kinase 2 expression suppresses growth and induces apoptosis of human tumor cells in a p53-dependent manner. AB - c-Jun N-terminal kinase (JNK) plays a critical role in coordinating the cellular response to stress and has been implicated in regulating cell growth and transformation. To investigate the growth-regulatory functions of JNK1 and JNK2, we used specific antisense oligonucleotides (AS) to inhibit their expression. A survey of several human tumor cell lines revealed that JNKAS treatment markedly inhibited the growth of cells with mutant p53 status but not that of cells with normal p53 function. To further examine the influence of p53 on cell sensitivity to JNKAS treatment, we compared the responsiveness of RKO, MCF-7, and HCT116 cells with normal p53 function to that of RKO E6, MCF-7 E6, and HCT116 p53(-/-), which were rendered p53 deficient by different methods. Inhibition of JNK2 (and to a lesser extent JNK1) expression dramatically reduced the growth of p53 deficient cells but not that of their normal counterparts. JNK2AS-induced growth inhibition was correlated with significant apoptosis. JNK2AS treatment induced the expression of the cyclin-dependent kinase inhibitor p21(Cip1/Waf1) in parental MCF-7, RKO, and HCT116 cells but not in the p53-deficient derivatives. That p21(Cip1/Waf1) expression contributes to the survival of JNK2AS-treated cells was supported by additional experiments demonstrating that p21(Cip1/Waf1) deficiency in HCT116 cells also results in heightened sensitivity to JNKAS treatment. Our results indicate that perturbation of JNK2 expression adversely affects the growth of otherwise nonstressed cells. p53 and its downstream effector p21(Cip1/Waf1) are important in counteracting these detrimental effects and promoting cell survival. PMID- 10669749 TI - Overexpression of kinase-associated phosphatase (KAP) in breast and prostate cancer and inhibition of the transformed phenotype by antisense KAP expression. AB - Accumulating evidence suggests that phosphatases play an important role in regulating a variety of signal transduction pathways that have a bearing on cancer. The kinase-associated phosphatase (KAP) is a human dual-specificity protein phosphatase that was identified as a Cdc2- or Cdk2-interacting protein by a yeast two-hybrid screening, yet the biological significance of these interactions remains elusive. We have identified the KAP gene as an overexpressed gene in breast and prostate cancer by using a phosphatase domain-specific differential-display PCR strategy. Here we report that breast and prostate malignancies are associated with high levels of KAP expression. The sublocalization of KAP is variable. In normal cells, KAP is primarily found in the perinuclear region, but in tumor cells, a significant portion of KAP is found in the cytoplasm. Blocking KAP expression by antisense KAP in a tetracycline regulatable system results in a reduced population of S-phase cells and reduced Cdk2 kinase activity. Furthermore, lowering KAP expression led to inhibition of the transformed phenotype, with reduced anchorage-independent growth and tumorigenic potential in athymic nude mice. These findings suggest that therapeutic intervention might be aimed at repression of KAP gene overexpression in human breast and prostate cancer. PMID- 10669750 TI - A combinatorial code for gene expression generated by transcription factor Bach2 and MAZR (MAZ-related factor) through the BTB/POZ domain. AB - Bach2 is a B-cell- and neuron-specific transcription repressor that forms heterodimers with the Maf-related oncoproteins. We show here that Bach2 activates transcription by interacting with its novel partner MAZR. MAZR was isolated by the yeast two-hybrid screen using the BTB/POZ domain of Bach2 as bait. Besides the BTB/POZ domain, MAZR possesses Zn finger motifs that are closely related to those of the Myc-associated Zn finger (MAZ) protein. MAZR mRNA was coexpressed with Bach2 in B cells among hematopoietic cells and in developing mouse limb buds, suggesting a cooperative role for MAZR and Bach2 in these cells. MAZR forms homo- and hetero-oligomers with Bach2 through the BTB domain, which oligomers bind to guanine-rich sequences. Unlike MAZ, MAZR functioned as a strong activator of the c-myc promoter in transfection assays with B cells. However, it does not possess a typical activation domain, suggesting a role for it as an unusual type of transactivator. The fgf4 gene, which regulates morphogenesis of limb buds, contains both guanine-rich sequences and a Bach2 binding site in its regulatory region. In transfection assays using fibroblast cells, the fgf4 gene was upregulated in the presence of both MAZR and Bach2 in a BTB/POZ domain-dependent manner. The results provide a new perspective on the function of BTB/POZ domain factors and indicate that BTB/POZ domain-mediated oligomers of transcription factors may serve as combinatorial codes for gene expression. PMID- 10669751 TI - Multiple mitogen-activated protein kinase signaling pathways connect the cot oncoprotein to the c-jun promoter and to cellular transformation. AB - The serine/threonine kinase Cot is a member of the mitogen-activated protein kinase (MAPK) kinase kinase family implicated in cellular transformation. Enhanced expression of this protein has been shown to activate both the MAPK and the c-Jun N-terminal kinase (JNK) pathways and to stimulate the nuclear factor of activated T cells and NF-kappaB-dependent transcription. However, the nature of the normal functions of the Cot protein and the molecular mechanisms responsible for its oncogenic potential are still largely unknown. Here, we show that overexpression of the cot proto-oncogene is sufficient to stimulate the expression of c-jun and that, in turn, the activity of c-Jun is required for Cot induced transformation. These observations prompted us to explore the molecular events by which Cot regulates c-jun expression. We found that Cot potently stimulates the activity of the c-jun promoter utilizing JNK-dependent and independent pathways, the latter involving two novel members of the MAPK family, p38gamma (ERK6) and ERK5. Molecularly, this activity was found to be dependent on the ability of Cot to activate, in vivo, members of each class of the MAPK kinase superfamily, including MEK, SEK, MKK6, and MEK5. Furthermore, the use of dominant interfering molecules revealed that Cot requires JNK, p38s, and ERK5 to stimulate the c-jun promoter fully and to induce neoplastic transformation. These findings indicate that Cot represents the first example of a serine/threonine kinase acting simultaneously on all known MAPK cascades. Moreover, these observations strongly suggest that the transforming ability of Cot results from the coordinated activation of these pathways, which ultimately converge on the regulation of the expression and activity of the product of the c-jun proto oncogene. PMID- 10669752 TI - Sequential requirements of the N-terminal palmitoylation site and SH2 domain of Src family kinases in the initiation and progression of FcepsilonRI signaling. AB - Initial biochemical signaling originating from high-affinity immunoglobulin E receptor (FcepsilonRI) has been ascribed to Src family kinases. To understand the mechanisms by which individual kinases drive the signaling, we conducted reconstitution experiments: FcepsilonRI signaling in RBL2H3 cells was first suppressed by a membrane-anchored, gain-of-function C-terminal Src kinase and then reconstructed with Src family kinases whose C-terminal negative regulatory sequence was replaced with a c-myc epitope. Those constructs derived from Lyn and Fyn, which are associated with detergent-resistant membranes (DRMs), physically interacted with resting FcepsilonRI and reconstructed clustering-induced signaling that leads to calcium mobilization and ERK1 and -2 activation. c-Src derived construct, which was excluded from DRMs, failed to interact with FcepsilonRI and to restore the signaling, whereas creation of palmitoylatable Cys3 enabled it to interact with DRMs and with FcepsilonRI and to restore the signaling. Deletion of Src homology 3 (SH3) domain from the Lyn-derived construct did not alter its ability to transduce the series of signaling. Deletion of SH2 domain did not affect its association with DRMs and with FcepsilonRI nor clustering-induced tyrosine phosphorylation of FcepsilonRI beta and gamma subunits, but it almost abrogated the next step of tyrosine phosphorylation of Syk and its recruitment to FcepsilonRI. These findings suggest that Lyn and Fyn could, but c-Src could not, drive FcepsilonRI signaling and that N-terminal palmitoylation and SH2 domain are required in sequence for the initial interaction with FcepsilonRI and for the signal progression to the molecular assembly. PMID- 10669753 TI - Mouse A6/twinfilin is an actin monomer-binding protein that localizes to the regions of rapid actin dynamics. AB - In our database searches, we have identified mammalian homologues of yeast actin binding protein, twinfilin. Previous studies suggested that these mammalian proteins were tyrosine kinases, and therefore they were named A6 protein tyrosine kinase. In contrast to these earlier studies, we did not find any tyrosine kinase activity in our recombinant protein. However, biochemical analysis showed that mouse A6/twinfilin forms a complex with actin monomer and prevents actin filament assembly in vitro. A6/twinfilin mRNA is expressed in most adult tissues but not in skeletal muscle and spleen. In mouse cells, A6/twinfilin protein is concentrated to the areas at the cell cortex which overlap with G-actin-rich actin structures. A6/twinfilin also colocalizes with the activated forms of small GTPases Rac1 and Cdc42 to membrane ruffles and to cell-cell contacts, respectively. Furthermore, expression of the activated Rac1(V12) in NIH 3T3 cells leads to an increased A6/twinfilin localization to nucleus and cell cortex, whereas a dominant negative form of Rac1(V12,N17) induces A6/twinfilin localization to cytoplasm. Taken together, these studies show that mouse A6/twinfilin is an actin monomer-binding protein whose localization to cortical G actin-rich structures may be regulated by the small GTPase Rac1. PMID- 10669754 TI - Sequestration and inhibition of Daxx-mediated transcriptional repression by PML. AB - PML fuses with retinoic acid receptor alpha (RARalpha) in the t(15;17) translocation that causes acute promyelocytic leukemia (APL). In addition to localizing diffusely throughout the nucleoplasm, PML mainly resides in discrete nuclear structures known as PML oncogenic domains (PODs), which are disrupted in APL and spinocellular ataxia cells. We isolated the Fas-binding protein Daxx as a PML-interacting protein in a yeast two-hybrid screen. Biochemical and immunofluorescence analyses reveal that Daxx is a nuclear protein that interacts and colocalizes with PML in the PODs. Reporter gene assay shows that Daxx drastically represses basal transcription, likely by recruiting histone deacetylases. PML, but not its oncogenic fusion PML-RARalpha, inhibits the repressor function of Daxx. In addition, SUMO-1 modification of PML is required for sequestration of Daxx to the PODs and for efficient inhibition of Daxx mediated transcriptional repression. Consistently, Daxx is found at condensed chromatin in cells that lack PML. These data suggest that Daxx is a novel nuclear protein bearing transcriptional repressor activity that may be regulated by interaction with PML. PMID- 10669755 TI - Arrest of G(1)-S progression by the p53-inducible gene PC3 is Rb dependent and relies on the inhibition of cyclin D1 transcription. AB - The p53-inducible gene PC3 (TIS21, BTG2) is endowed with antiproliferative activity. Here we report that expression of PC3 in cycling cells induced accumulation of hypophosphorylated, growth-inhibitory forms of pRb and led to G(1) arrest. This latter was not observed in cells with genetic disruption of the Rb gene, indicating that the PC3-mediated G(1) arrest was Rb dependent. Furthermore, (i) the arrest of G(1)-S transition exerted by PC3 was completely rescued by coexpression of cyclin D1 but not by that of cyclin A or E; (ii) expression of PC3 caused a significant down-regulation of cyclin D1 protein levels, also in Rb-defective cells, accompanied by inhibition of CDK4 activity in vivo; and (iii) the removal from the PC3 molecule of residues 50 to 68, a conserved domain of the PC3/BTG/Tob gene family, which we term GR, led to a loss of the inhibition of proliferation as well as of the down-regulation of cyclin D1 levels. These data point to cyclin D1 down-regulation as the main factor responsible for the growth inhibition by PC3. Such an effect was associated with a decrease of cyclin D1 transcript and of cyclin D1 promoter activity, whereas no effect of PC3 was observed on cyclin D1 protein stability. Taken together, these findings indicate that PC3 impairs G(1)-S transition by inhibiting pRb function in consequence of a reduction of cyclin D1 levels and that PC3 acts, either directly or indirectly, as a transcriptional regulator of cyclin D1. PMID- 10669756 TI - A DNA helicase required for maintenance of the functional mitochondrial genome in Saccharomyces cerevisiae. AB - A novel DNA helicase, a homolog of several prokaryotic helicases, including Escherichia coli Rep and UvrD proteins, is encoded by the Saccharomyces cerevisiae nuclear genome open reading frame YOL095c on the chromosome XV. Our data demonstrate that the helicase is localized in the yeast mitochondria and is loosely associated with the mitochondrial inner membrane during biochemical fractionation. The sequence of the C-terminal end of the 80-kDa helicase protein is similar to a typical N-terminal mitochondrial targeting signal; deletions and point mutations in this region abolish transport of the protein into mitochondria. The C-terminal signal sequence of the helicase targets a heterologous carrier protein into mitochondria in vivo. The purified recombinant protein can unwind duplex DNA molecules in an ATP-dependent manner. The helicase is required for the maintenance of the functional ([rho(+)]) mitochondrial genome on both fermentable and nonfermentable carbon sources. However, the helicase is not essential for the maintenance of several defective ([rho(-)]) mitochondrial genomes. We also demonstrate that the helicase is not required for transcription in mitochondria. PMID- 10669757 TI - Calcium depletion dissociates and activates heterodimeric notch receptors. AB - Notch receptors participate in a highly conserved signaling pathway that regulates morphogenesis in multicellular animals. Maturation of Notch receptors requires the proteolytic cleavage of a single precursor polypeptide to produce a heterodimer composed of a ligand-binding extracellular domain (N(EC)) and a single-pass transmembrane signaling domain (N(TM)). Notch signaling has been correlated with additional ligand-induced proteolytic cleavages, as well as with nuclear translocation of the intracellular portion of N(TM) (N(ICD)). In the current work, we show that the N(EC) and N(TM) subunits of Drosophila Notch and human Notch1 (hN1) interact noncovalently. N(EC)-N(TM) interaction was disrupted by 0.1% sodium dodecyl sulfate or divalent cation chelators such as EDTA, and stabilized by millimolar Ca(2+). Deletion of the Ca(2+)-binding Lin12-Notch (LN) repeats from the N(EC) subunit resulted in spontaneous shedding of N(EC) into conditioned medium, implying that the LN repeats are important in maintaining the interaction of N(EC) and N(TM). The functional consequences of EDTA-induced N(EC) dissociation were studied by using hN1-expressing NIH 3T3 cells. Treatment of these cells for 10 to 15 min with 0.5 to 10 mM EDTA resulted in the rapid shedding of N(EC), the transient appearance of a polypeptide of the expected size of N(ICD), increased intranuclear anti-Notch1 staining, and the transient activation of an Notch-sensitive reporter gene. EDTA treatment of HeLa cells expressing endogenous Notch1 also stimulated reporter gene activity to a degree equivalent to that resulting from exposure of the cells to the ligand Delta1. These findings indicate that receptor activation can occur as a consequence of N(EC) dissociation, which relieves inhibition of the intrinsically active N(TM) subunit. PMID- 10669758 TI - The neuron-enriched splicing pattern of Drosophila erect wing is dependent on the presence of ELAV protein. AB - Although the Drosophila melanogaster erect wing (ewg) gene is broadly transcribed in adults, an unusual posttranscriptional regulation involving alternative and inefficient splicing generates a 116-kDa EWG protein in neurons, while protein expression elsewhere or of other isoforms is below detection at this stage. This posttranscriptional control is important, as broad expression of EWG can be lethal. In this paper, we show that ELAV, a neuron-specific RNA binding protein, is necessary to regulate EWG protein expression in ELAV-null eye imaginal disc clones and that ELAV is sufficient for EWG expression in wing disc imaginal tissue after ectopic expression. Further, analysis of EWG expression elicited from intron-containing genomic transgenes and cDNA minitransgenes in ELAV deficient eye discs shows that this regulation is dependent on the presence of ewg introns. Analyses of the ewg splicing patterns in wild-type and ELAV deficient eye imaginal discs and in wild-type and ectopic ELAV-expressing wing imaginal discs, show that certain neuronal splice isoforms correspond to ELAV levels. The data presented in this paper are consistent with a mechanism in which ELAV increases the splicing efficiency of ewg transcripts in alternatively spliced regions rather than with a mechanism in which stability of specific splice forms is enhanced by ELAV. Additionally, we report that ELAV promotes a neuron-enriched splice isoform of Drosophila armadillo transcript. ELAV, however, is not involved in all neuron-enriched splice events. PMID- 10669759 TI - Molecular cloning of apobec-1 complementation factor, a novel RNA-binding protein involved in the editing of apolipoprotein B mRNA. AB - The C-to-U editing of apolipoprotein B (apo-B) mRNA is catalyzed by a multiprotein complex that recognizes an 11-nucleotide mooring sequence downstream of the editing site. The catalytic subunit of the editing enzyme, apobec-1, has cytidine deaminase activity but requires additional unidentified proteins to edit apo-B mRNA. We purified a 65-kDa protein that functionally complements apobec-1 and obtained peptide sequence information which was used in molecular cloning experiments. The apobec-1 complementation factor (ACF) cDNA encodes a novel 64.3 kDa protein that contains three nonidentical RNA recognition motifs. ACF and apobec-1 comprise the minimal protein requirements for apo-B mRNA editing in vitro. By UV cross-linking and immunoprecipitation, we show that ACF binds to apo B mRNA in vitro and in vivo. Cross-linking of ACF is not competed by RNAs with mutations in the mooring sequence. Coimmunoprecipitation experiments identified an ACF-apobec-1 complex in transfected cells. Immunodepletion of ACF from rat liver extracts abolished editing activity. The immunoprecipitated complexes contained a functional holoenzyme. Our results support a model of the editing enzyme in which ACF binds to the mooring sequence in apo-B mRNA and docks apobec 1 to deaminate its target cytidine. The fact that ACF is widely expressed in human tissues that lack apobec-1 and apo-B mRNA suggests that ACF may be involved in other RNA editing or RNA processing events. PMID- 10669760 TI - Mouse Zac1, a transcriptional coactivator and repressor for nuclear receptors. AB - Transcriptional activation by nuclear hormone receptors is mediated by the 160 kDa family of nuclear receptor coactivators. These coactivators associate with DNA-bound nuclear receptors and transmit activating signals to the transcription machinery through two activation domains. In screening for mammalian proteins that bind the C-terminal activation domain of the nuclear receptor coactivator GRIP1, we identified a new variant of mouse Zac1 which we call mZac1b. Zac1 was previously discovered as a putative transcriptional activator involved in regulation of apoptosis and the cell cycle. In yeast two-hybrid assays and in vitro, mZac1b bound to GRIP1, to CREB-binding protein (CBP) and p300 (which are coactivators for nuclear receptors and other transcriptional activators), and to nuclear receptors themselves in a hormone-independent manner. In transient transfection assays mZac1b exhibited a transcriptional activation activity when fused with the Gal4 DNA binding domain, and it enhanced transcriptional activation by the Gal4 DNA binding domain fused to GRIP1 or CBP fragments. More importantly, mZac1b was a powerful coactivator for the hormone-dependent activity of nuclear receptors, including androgen, estrogen, glucocorticoid, and thyroid hormone receptors. However, with some reporter genes and in some cell lines mZac1b acted as a repressor rather than a coactivator of nuclear receptor activity. Thus, mZac1b can interact with nuclear receptors and their coactivators and play both positive and negative roles in regulating nuclear receptor function. PMID- 10669761 TI - The coactivator PGC-1 cooperates with peroxisome proliferator-activated receptor alpha in transcriptional control of nuclear genes encoding mitochondrial fatty acid oxidation enzymes. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) plays a key role in the transcriptional control of genes encoding mitochondrial fatty acid beta oxidation (FAO) enzymes. In this study we sought to determine whether the recently identified PPAR gamma coactivator 1 (PGC-1) is capable of coactivating PPARalpha in the transcriptional control of genes encoding FAO enzymes. Mammalian cell cotransfection experiments demonstrated that PGC-1 enhanced PPARalpha mediated transcriptional activation of reporter plasmids containing PPARalpha target elements. PGC-1 also enhanced the transactivation activity of a PPARalpha Gal4 DNA binding domain fusion protein. Retroviral vector-mediated expression studies performed in 3T3-L1 cells demonstrated that PPARalpha and PGC-1 cooperatively induced the expression of PPARalpha target genes and increased cellular palmitate oxidation rates. Glutathione S-transferase "pulldown" studies revealed that in contrast to the previously reported ligand-independent interaction with PPARgamma, PGC-1 binds PPARalpha in a ligand-influenced manner. Protein-protein interaction studies and mammalian cell hybrid experiments demonstrated that the PGC-1-PPARalpha interaction involves an LXXLL domain in PGC 1 and the PPARalpha AF2 region, consistent with the observed ligand influence. Last, the PGC-1 transactivation domain was mapped to within the NH(2)-terminal 120 amino acids of the PGC-1 molecule, a region distinct from the PPARalpha interacting domains. These results identify PGC-1 as a coactivator of PPARalpha in the transcriptional control of mitochondrial FAO capacity, define separable PPARalpha interaction and transactivation domains within the PGC-1 molecule, and demonstrate that certain features of the PPARalpha-PGC-1 interaction are distinct from that of PPARgamma-PGC-1. PMID- 10669762 TI - The BIR motifs mediate dominant interference and oligomerization of inhibitor of apoptosis Op-IAP. AB - The defining structural motif of the inhibitor of apoptosis (iap) protein family is the BIR (baculovirus iap repeat), a highly conserved zinc coordination domain of approximately 70 residues. Although the BIR is required for inhibitor-of apoptosis (IAP) function, including caspase inhibition, its molecular role in antiapoptotic activity in vivo is unknown. To define the function of the BIRs, we investigated the activity of these structural motifs within Op-IAP, an efficient, virus-derived IAP. We report here that Op-IAP(1-216), a loss-of-function truncation which contains two BIRs but lacks the C-terminal RING motif, potently interfered with Op-IAP's capacity to block apoptosis induced by diverse stimuli. In contrast, Op-IAP(1-216) had no effect on apoptotic suppression by caspase inhibitor P35. Consistent with a mechanism of dominant inhibition that involves direct interaction between Op-IAP(1-216) and full-length Op-IAP, both proteins formed an immunoprecipitable complex in vivo. Op-IAP also self-associated. In contrast, the RING motif-containing truncation Op-IAP(183-268) failed to interact with or interfere with Op-IAP function. Substitution of conserved residues within BIR 2 caused loss of dominant inhibition by Op-IAP(1-216) and coincided with loss of interaction with Op-IAP. Thus, residues encompassing the BIRs mediate dominant inhibition and oligomerization of Op-IAP. Consistent with dominant interference by interaction with an endogenous cellular IAP, Op-IAP(1-216) also lowered the survival threshold of cultured insect cells. Taken together, these data suggest a new model wherein the antiapoptotic function of IAP requires homo oligomerization, which in turn mediates specific interactions with cellular apoptotic effectors. PMID- 10669763 TI - Posttranslational modification of Bcl-2 facilitates its proteasome-dependent degradation: molecular characterization of the involved signaling pathway. AB - The ratio of proapoptotic versus antiapoptotic Bcl-2 members is a critical determinant that plays a significant role in altering susceptibility to apoptosis. Therefore, a reduction of antiapoptotic protein levels in response to proximal signal transduction events may switch on the apoptotic pathway. In endothelial cells, tumor necrosis factor alpha (TNF-alpha) induces dephosphorylation and subsequent ubiquitin-dependent degradation of the antiapoptotic protein Bcl-2. Here, we investigate the role of different putative phosphorylation sites to facilitate Bcl-2 degradation. Mutation of the consensus protein kinase B/Akt site or of potential protein kinase C or cyclic AMP dependent protein kinase sites does not affect Bcl-2 stability. In contrast, inactivation of the three consensus mitogen-activated protein (MAP) kinase sites leads to a Bcl-2 protein that is ubiquitinated and subsequently degraded by the 26S proteasome. Inactivation of these sites within Bcl-2 revealed that dephosphorylation of Ser87 appears to play a major role. A Ser-to-Ala substitution at this position results in 50% degradation, whereas replacement of Thr74 with Ala leads to 25% degradation, as assessed by pulse-chase studies. We further demonstrated that incubation with TNF-alpha induces dephosphorylation of Ser87 of Bcl-2 in intact cells. Furthermore, MAP kinase triggers phosphorylation of Bcl-2, whereas a reduction in Bcl-2 phosphorylation was observed in the presence of MAP kinase-specific phosphatases or the MAP kinase-specific inhibitor PD98059. Moreover, we show that oxidative stress mediates TNF-alpha-stimulated proteolytic degradation of Bcl-2 by reducing MAP kinase activity. Taken together, these results demonstrate a direct protective role for Bcl-2 phosphorylation by MAP kinase against apoptotic challenges to endothelial cells and other cells. PMID- 10669764 TI - Insect cells as HLA-restricted antigen-presenting cells for the IFN-gamma elispot assay. AB - Measurement of specific cellular immune responses in patients undergoing immunotherapy is difficult. Established approaches, including cytotoxicity (e.g., 51Cr release) and cytokine release assays, require in vitro culturing for several weeks or more of patients' peripheral blood mononuclear cells (PBMC) and the addition of exogenous cytokines. Therefore, the immunological response does not reflect in vivo conditions. To address these disadvantages, we have used an interferon-gamma (IFN-gamma) Elispot assay for detecting peptide-specific CD8(+) lymphocytes in PBMC. A limitation of this assay is the lack of a reproducible source of antigen-presenting cells (APCs). Currently available APCs often lead to significant background levels. It has been shown that transfected insect cells can express empty MHC class I molecules on their surface. We have transfected Drosophila melanogaster S2 cells and the Lepidopteran line Sf9 with the gene encoding human HLA-A2.1. We demonstrate that insect cells expressing a human HLA molecule effectively function as APCs in the IFN-gamma Elispot assay. Initially the feasibility of the assay was assessed using CD8(+) T cells from HLA-A2.1(+) donors with known reactivity against an HLA-A2.1-binding epitope of the influenza matrix protein. Use of insect cells as APCs abrogated background spots, increasing sensitivity. We further observed that a short-term prestimulation of PBMC with peptide-pulsed insect cells markedly enhanced the frequency of peptide specific T cells that could be measured in the Elispot assay without increasing the background. This approach was then used to measure CD8(+) T cell reactivity to a peptide from tyrosinase, an antigen that is processed and presented by melanoma cells. Insect cells expressing human HLA molecules provide a standard APC for monitoring CD8(+) T cell responses to tumor and viral peptides during immunotherapy. PMID- 10669765 TI - Measurement of T-helper cytokines secreted by cord blood mononuclear cells in response to allergens. AB - It has been proposed that in utero factors may predispose towards the development of childhood atopy. To test this hypothesis, it will be necessary to measure T helper cell (Th) cytokines secreted by human cord blood mononuclear cells (CBMC) stimulated by allergens. However, to date, it has proven impossible to measure allergen-specific CBMC secretion of the key Th cytokine interleukin-4 (IL-4) using conventional sandwich ELISA techniques. We report for the first time the successful measurement of IL-4 secreted by CBMC stimulated by the allergens timothy grass pollen and house dust mite extract. The method is an adaptation of a novel cell-based ELISA (celELISA), which demonstrated an increased (up to 20 fold) sensitivity to detect IL-4. The method is simple, precise, is no more costly than a conventional ELISA, and can identify individuals in a general population whose CBMC exhibit different cytokine biases in response to allergens. The frequency distribution of IL-4 and interferon-gamma (IFN-gamma) CBMC responses to allergens in the general population approximates to a log-normal distribution, which will permit the application of linear regression techniques in the identification of in utero factors which influence Th bias. PMID- 10669766 TI - Use of linear and multiple antigenic peptides in the immunodiagnosis of acute hepatitis A virus infection. AB - The reactivities of two panels of anti-HAV human sera from geographically distinct areas (Chile and Spain) to synthetic peptides from the VP1, VP2 and VP3 hepatitis A virus capsid proteins were examined by an enzyme-linked immunosorbent assay (ELISA) procedure. Two and four branched multiple antigenic peptides (MAPs) and palmitoylated peptides were compared with free synthetic sequences for the detection of IgM anti-HAV antibodies in the two panels of human sera. Our results showed that acute hepatitis A patient sera recognized preferentially homogeneous two branched MAPs and palmitic acid conjugated peptides. The palmitoyl-derived VP3(110-121) peptide and the corresponding dimeric MAP were the most sensitive and appropriate for serological studies of HAV-infected patients by ELISA, sensitivity and specificity being higher than 90% and 95%, respectively. These peptide-based tests open up new avenues in the development of peptide-based immunosorbent assays for the detection of acute HAV disease. PMID- 10669767 TI - A low budget luminometer for sensitive chemiluminescent immunoassays. AB - We have designed a simple luminometer based on a reasonably priced Peltier-cooled charge-coupled device (CCD) camera, housed in a light-tight box, with straightforward lens imaging and a simple platform for a microtitre or other assay format. The quantitative readout of the CCD image is recorded on a PC using customised software. The instrument can be assembled in a standard university workshop for under pound3000, compared with the cheapest commercial instruments retailing at pound10,000 and above. Consistent with the general view on chemiluminescent assays, the sensitivity is 10-100 times greater than that obtained with parallel ELISA's using a chromogenic substrate. A unique feature of the CCD format is that it enables assays to be carried out on arrays of minidots and even nanodots of antigen on the floor of each microtitre well. This permits direct comparison and standardisation of reactivity of a single sample against several antigens and economy in the use of reagents, test sample and technician time; finger-prick samples of blood can be analysed. The instrument should have widespread applicability in developing countries and, indeed, in any laboratories with hard-pressed budgets. PMID- 10669768 TI - Zinc has no effect on IL-3-mediated apoptosis of BAF-3 cells but enhances CD95 mediated apoptosis of jurkat cells. AB - The feasibility of using a zinc-inducible gene expression system for the study of apoptosis-controlling genes in BAF-3 murine B cells and Jurkat human T cells was evaluated. Initially, cell sensitivity to a range of zinc concentrations was examined. It was found that zinc concentrations above 60 microM were toxic to BAF 3 cells and those above 50 microM were toxic to Jurkat cells. Secondly, the zinc concentration required to achieve maximal gene expression was examined. BAF-3 cells transiently transfected with the pMTCB6+/luciferase vector were exposed to zinc concentrations ranging from 0-120 microM, whilst stably transfected Jurkat cells were exposed to 0-70 microM zinc. At zinc concentrations nontoxic to each cell type, the maximum induction achieved was 20-fold (at 60 microM) in BAF-3 cells, and 7.5-fold (at 50 microM) in Jurkat cells. Thirdly, the effect of zinc on apoptosis was examined. It was shown that exposure to nontoxic zinc concentrations had no effect on IL-3 withdrawal-mediated apoptosis of BAF-3 cells. However, in the case of Jurkat cells, pre-exposure to zinc augmented CD95 mediated apoptosis. These results illustrate the importance of characterizing individual cell lines when using zinc-inducible gene expression systems. PMID- 10669769 TI - Development of a new method for the determination of immune responses in the human stomach. AB - The discovery of the gastric pathogen Helicobacter pylori has created a need for accurate methods to study immune responses locally in the human stomach. Therefore, we have developed a quick and easy method for extraction of antibodies from gastric biopsies using saponin and compared this method with the more laborious analysis of antibody-secreting cells (ASCs) from gastric biopsies. We have also analyzed the antibody content in gastric aspirates, saliva and plasma. There was a strong correlation between the total IgA levels in the biopsy extracts and the frequencies of IgA-secreting cells. In addition, the IgA and IgG levels against a H. pylori whole membrane preparation and purified urease in the biopsy extracts correlated well with the frequencies of specific IgA and IgG secreting cells. However, the antibody levels in gastric aspirates, saliva and plasma specimens did not correlate with the frequencies of corresponding ASC in the gastric biopsies. Thus, the saponin extraction method is suitable for monitoring local antibody responses in the stomach, while analyses of gastric aspirates, saliva or plasma are not appropriate for this purpose. PMID- 10669770 TI - A simplified procedure for the preparation of MHC/peptide tetramers: chemical biotinylation of an unpaired cysteine engineered at the C-terminus of MHC-I. AB - Recently, a powerful approach for the detection of MHC/peptide-specific T cells has been made possible by the engineering of soluble-tetrameric MHC/peptide complexes, consisting of singly biotinylated MHC/peptide molecules bound to fluorescent-labeled streptavidin. These tetrameric molecules are thought to compensate for the low affinity and relative fast dissociation rate of the TCR/MHC-peptide interaction by increasing the avidity of this interaction, thus allowing the stable binding of MHC/peptide tetramers to TCR expressing cells. Here we describe a new more simplified procedure for obtaining MHC/peptide tetramers using the well-characterized H-2K(b)/VSV system. This procedure consists of the incorporation of an unpaired cysteine residue at the C-terminus of the H-2K(b) molecule, allowing site-specific biotinylation by a -SH-specific biotinylating reagent. The H-2K(b)/VSV tetramers bound only to hybridomas expressing H-2K(b)/VSV-specific TCRs. When coated on a plate, these tetramers were able to induce IL-2 release by those hybridomas. Furthermore, H-2K(b)/VSV tetramers bound to CTL populations obtained from mice immunized with VSV-peptide. The specificity of the binding was further refined by studying cross-recognition of VSV by CTL populations obtained from mice immunized with single amino acid substituted VSV peptide variants. H-2K(b)/VSV tetramers bound only to those CTL populations that cross-reacted with the wild-type VSV peptide. Our method provides a simple, efficient and inexpensive procedure for making MHC/peptide tetramers, a highly specific and very useful reagent with a number of important applications in basic and clinical T cell research. PMID- 10669771 TI - High-performance sodium dodecyl sulfate-capillary gel electrophoresis of antibodies and antibody fragments. AB - High-performance sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) has been used to separate antibodies and their fragments according to size. Under non-reduced conditions, murine monoclonal antibodies generally show a predominant peak with five to six apparent fragment peaks. The magnitude of the apparent fragmentation is temperature-dependent and is more pronounced with rabbit, sheep and bovine immunoglobulin G's than murine monoclonal antibodies. In addition to temperature, pH and buffer also affect the fragmentation. Without heat treatment during the preparation of the SDS-antibody complexes, the observed fragments become nearly absent; however, some murine monoclonal antibodies exhibit several peaks that group near the expected migration time of an immunoglobulin G, presumably due to their anomalous interaction with sodium dodecyl sulfate. The method can also be used to monitor the progress of peptic digestions to generate murine F(ab')(2) antibody, to perform "gel-shift" assays, and to separate mouse ascitic fluid. This high-performance electrophoretic technique is suitable for quality control as well as the characterization of the antibodies under experimental conditions. PMID- 10669772 TI - Perfusion chromatography for very rapid purification of class I and II MHC proteins. AB - Major histocompatibility complex (MHC) proteins are surface glycoproteins that are strongly associated with either self or foreign peptides. Their interaction with the T-cell receptor on the T-cells initiates an immune response and help in discriminating between self and non-self, respectively. We describe here a novel means of rapidly purifying human MHC molecules on either small scale or large scale from the cell lysate of lymphoblastoid B cell line and from insect cell culture supernatants by using affinity perfusion chromatography. As representative cases HLA-B2705, a class I MHC molecule, and HLA-DR1, a class II MHC molecule were purified from EBV-transformed human lymphoblastoid B cells, LG2. Soluble HLA-DR1 was also purified from the cell culture supernatant of insect cells. The peptides eluted from the purified HLA-B2705 were pool sequenced and found to have the same motif as has previously been published. This new method provides a very rapid means of purifying MHC protein molecules, applicable to both large scale and small scale purification, which in turn greatly enhances the accuracy of further analysis of the associated peptides through mass spectrometry. PMID- 10669773 TI - Comparative study of two cytofluorometric methods of analysis. AB - Cytofluorometry of lymphocytes is an important technique in experimental biology and clinical medicine. One source of variability in the results with this technique stems from the difficulty of delineating cell subpopulations on a visual basis. We have evaluated the performance of a novel software method (Attractor), which introduces cluster analysis for the more precise definition of cell populations. Using 115 blood samples from patients with various immunological diseases, we compared the results obtained for 19 lymphocyte cell populations employing either the Attractor program or a conventional program (Cellquest). The analysis focused on inter-observer and inter-method variability and comparability. Inter-observer variability was significantly lower with the Attractor software, particularly when quantifying small cell populations such as activated subsets. The results obtained with both methods showed high correlation coefficients except for some cell populations that were either very small or which had to be calculated from the sum of other counts. The performance of the novel flow cytometric software is similar to software programs currently in use, but it offers an advantage for the definition of small and/or activated lymphocyte subpopulations. Moreover, the consistency of the measurements is better. A major disadvantage for statistical analysis, however, is that the Attractor program has not been adapted for non-parametric data. PMID- 10669774 TI - ELISA for a complexed antigen with a monoclonal antibody blocking reaction with the free antigen-assay-specific for complexed prostate-specific antigen. AB - We have developed an enzyme-linked immunoassay (ELISA) for serum complexed antigen (prostate-specific antigen; PSA)(c-PSA) with simultaneous blocking of free-antigen (PSA)(f-PSA). The assay utilizes three different monoclonal antibodies (MAbs) recognising three distinct PSA epitopes. The detection limit was established as 0.19 microg/l (n=20, mean of zero standard+2S.D.) and the average recovery of f-PSA was 98-100%. The within-run and between-day coefficients of variation (CV) ranged from 2.1% to 3.2% and 2.8% to 6.3%, respectively. There was a good correlation between serum c-PSA measured by the present ELISA and PSA-alpha(1)-antichymotrypsin complex (PSA-ACT) concentrations (r=0.991). This method should provide a better tool for discriminating between benign and malignant prostatic disease. PMID- 10669776 TI - Identification of human TGF-beta1 signal (leader) sequence polymorphisms by PCR RFLP. AB - Links between disease susceptibility and genetically determined variation in human cytokine expression have recently been described. This has led to a demand for simple methods of identifying cytokine gene polymorphisms of potential clinical relevance. Here, we describe a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying two human transforming growth factor beta1 (TGF-beta1) signal (leader) sequence polymorphisms, T869C (Leu10Pro) and G915C (Arg25Pro). This permits simple and robust identification of TGF-beta1 leader sequence genotypes and demonstrates the physical linkage in cis between T869C (Leu10Pro) and G915C (Arg25Pro). The method does not require previously genotyped standards. The efficacy of enzyme digestion is internally controlled by the presence of conserved restriction sites. PMID- 10669775 TI - Generation of high-affinity rabbit polyclonal antibodies to the murine urokinase receptor using DNA immunization. AB - The urokinase receptor (uPAR) is a glycolipid anchored cell surface glycoprotein that plays a central role in extracellular proteolysis during tissue remodeling processes and cancer invasion. By intramuscular (i.m.) injection of rabbits with plasmid DNA coding for a carboxy-terminally truncated secreted form of the murine uPAR (muPAR), specific anti-sera with a titer of 64,000, as measured by ELISA, have been obtained. Rabbits received a total of 10 monthly injections of 1 mg DNA in phosphate-buffered saline. The antibody titer peaked between the 5th and 7th injection and slowly declined after the 8th injection. After the final immunization the immune response persisted for at least 6 months without further injections. The antibodies generated by DNA immunization were useful for immunohistochemistry and immunoblotting, recognizing the antigen both in its native and in its reduced and alkylated form. Using the antibodies in immunoblotting muPAR was identified in lysates of peritoneal macrophages, spleen and lung tissue. Both the intact and cleaved form of muPAR were identified in lysates of a murine monocyte cell line P388D.1. No cross-reaction with human uPAR was observed. In immunohistochemical analysis of normal mouse lung tissue uPAR immunoreactivity was located in the alveoli and pulmonary vessels, whereas the bronchial epithelium was negative. These results demonstrate that DNA immunization of rabbits using i.m. injection is a very effective and easy method to raise polyclonal antibodies which can be used for characterization and localization of muPAR in mouse tissue. PMID- 10669777 TI - Production of monoclonal antibodies using recombinant baculovirus displaying gp64 fusion proteins. AB - Generation of protein immunogens is often a rate-limiting step in the production of monoclonal antibodies (Mabs). Expressing domains of proteins as fusions to the baculovirus surface glycoprotein gp64 displays foreign proteins on the surface of the virion. Antigen is produced by inserting a gene fragment in-frame between the signal sequence and the mature protein domain of the gp64 nucleotide sequence. This method allows immunization with whole virus, eliminating the need for purification of target antigens. Affinity-matured Mabs to the human nuclear receptors LXRbeta and FXR have been produced using baculovirus particles displaying gp64/nuclear receptor fusion proteins as the immunizing agent. Immunizations were performed directly with pelleted virus using the Repetitive Immunization Multiple Sites (RIMMS) immunization strategy for rapid Mab production. All Mabs were identified using insect cells infected with the immunizing virus. Characterization of these antibodies shows them to be class switched and specific for LXRbeta or FXR. Additionally, high affinity antibodies that recognize gp64 and neutralize baculovirus infection of insect cells were isolated. Use of the recombinant baculovirus gp64 display system makes possible the production of Mabs once a partial DNA sequence is known. This allows the generation of antibodies prior to the isolation of purified protein, in turn providing antibodies to facilitate purification, characterization and immunolocalization of proteins. PMID- 10669778 TI - Transient expression of bacterial gene fragments in eukaryotic cells: implications for CD8(+) T cell epitope analysis. AB - CD8(+) T cells are potent effectors of acquired immunity against some viruses and intracellular bacterial pathogens. Antigens recognized by CD8(+) T cells are small, 8-9 amino acid peptides derived from proteins produced by the pathogen. These peptides are presented by MHC class I molecules on the surface of the infected cell. When characterizing the CD8(+) T cell response to a bacterial or viral pathogen, it is often necessary to express an antigenic protein in a eukaryotic host cell that is capable of processing and presenting peptide epitopes to antigen-specific CD8(+) T cells. We describe a system designed to transiently express bacterial polypeptides and MHC class I molecules in eukaryotic cells. Recognition of these peptide-MHC complexes stimulates TNF production by antigen-specific CD8(+) T cell lines. This system should be useful for analysis of CD8(+) T cell epitope-containing bacterial gene fragments when expression of the entire bacterial protein is detrimental to the eukaryotic cell, or when overexpression of the bacterial gene is detrimental to the bacterial cloning strain. Furthermore, this system can be used for the rapid mapping of CD8(+) T cell epitopes within a protein. PMID- 10669779 TI - A polymorphism in the mouse crg-2/IP-10 gene complicates chemokine gene expression analysis using a commercial ribonuclease protection assay. AB - The use of multiprobe RPAs is becoming an increasingly popular method for the detection and quantitation of RNA levels in cells and tissues. Here we report that due to a polymorphism in the 3'-noncoding region of the mouse Crg-2/IP-10 gene, the mCK-5 chemokine probe set available from Pharmingen can yield aberrant signal patterns with RNA samples from BALB/c, MRL and possibly other mouse strains that may lead to false conclusions regarding expression of the Crg-2/IP 10 and MCP-1 genes. PMID- 10669780 TI - Cell-ELISA using beta-galactosidase conjugated antibodies. AB - Cell-enzyme-linked immunosorbent assay (cell-ELISA) is a technique for the rapid, convenient, and quantitative detection of molecules expressed on the cell surface. Here we present an evaluation of beta-galactosidase as an antibody-tag for cell-ELISA. In contrast to substrates for horseradish peroxidase (HRP) and alkaline phosphatase, murine splenocytes do not hydrolyze the beta-galactosidase substrate chlorophenolred-beta-D-galactopyranoside (CPRG). beta-Galactosidase antibody conjugates show much lower background binding to murine T cells than conjugates with HRP or alkaline phosphatase. We describe step-by-step procedures for direct and indirect beta-galactosidase based cell-ELISA to quantitate the expression of molecules on the surface of unfixed, live cells. Variations of the basic protocol are suitable for adherent and non-adherent cells, large scale screening for expression of cell surface molecules, and the screening of hybridomas for production of antibodies to cell surface epitopes. Since relatively few beta-galactosidase conjugated antibodies are commercially available, we describe an efficient method to couple beta-galactosidase to antibodies using a novel water soluble heterobifunctional crosslinker, sulfosuccinimidyl 4-[N-maleimidomethyl]-cyclohexane-1-carboxylate (sulfo-SMCC). We demonstrate the utility of this method by conjugating F(ab')(2) fragments of an anti-B7-2 antibody, and using this conjugate to assay B7-2 on Fc-receptor bearing cells. PMID- 10669781 TI - The polyprotein lipid binding proteins of nematodes. AB - The nematode polyprotein allergens/antigens (NPAs) are specific to nematodes, and are synthesised as tandemly repetitive polypeptides comprising 10 or more repeated units. The polyproteins are post-translationally cleaved at consensus sites to yield multiple copies of the approximately 15-kDa NPA units. These units can be highly diverse in their amino acid sequences, but absolutely conserved signature amino acid positions are identifiable. NPA units are helix-rich and possibly fold as four helix bundle proteins. The NPA units have relatively non specific lipid binding activities, binding fatty acids and retinoids, with dissociation constants similar to those of lipid transport proteins of vertebrates. Fluorescence-based analysis has indicated that, like most lipid transport proteins, the ligand is taken into the binding site in its entirety, but the binding site environment is unusual. NPAs are synthesised in the gut of nematodes, and presumably act to distribute small lipids from the gut, via the pseudocoelomic fluid, to consuming tissues (muscles, gonads, etc.). In some species, one of the units has a histidine-rich extension peptide which binds haems and certain divalent metal ions. NPAs appear to be released by parasitic nematodes, and may thereby be involved in modification of the local inflammatory and immunological environment of the tissues they inhabit by delivering or sequestering pharmacologically active lipids - they are known to bind arachidonic acids and some of its metabolites, lysophospholipids, and retinoids. NPAs are the only known lipid binding protein made as polyproteins, and are exceptions to the rule that repetitive polyproteins are only produced by cells undergoing programmed cell death and producing specialist products. PMID- 10669782 TI - A conformational transition between an open and closed form of human pancreatic lipase revealed by a monoclonal antibody. AB - The interfacial activation of human pancreatic lipase (HPL) probably involves the motion of a lid covering the active site of the enzyme. Here we observed that the presence of either bile salts or a small proportion of water-miscible organic solvents (called activator compounds) considerably enhances the enzymatic activity of HPL on a monomeric solution of tripropionin. This finding suggests that the activator compounds may favor the opening of the lid. This hypothesis was checked by comparing the immunoreactivity of HPL and HPL with a mini-lid (HPL(-lid)) towards anti-HPL monoclonal antibodies (mAbs), in the presence and absence of the activator compounds. A single conformational mAb (248-31) fails to immunoprecipitate HPL in the presence of activator compounds and HPL covalently inhibited with diethyl p-nitrophenyl phosphate (DP.HPL). This loss of recognition of HPL by mAb 248-31 was probably due to the motion of the lid, since HPL(-lid) was always recognized in the presence or absence of activator compounds. Furthermore, two other mAbs (81-23 and 146-40) immunoprecipitated HPL similarly whether or not the activator compounds were present. MAb 248-31 therefore specifically recognizes HPL in the closed but not the open conformation. PMID- 10669783 TI - Tryptophanyl contributions to apomyoglobin fluorescence resolved by site-directed mutagenesis. AB - The individual emission properties of the two tryptophanyl residues of sperm whale apomyoglobin have been resolved by examining the fluorescence variations induced by denaturants, i.e., acid and guanidine, on apomyoglobin mutants W7F and W14F. The fluorescence changes have been correlated to the conformational transitions undergone by apomyoglobin on increasing denaturant concentration. The results indicate that the fluorescence decrease, observed for sperm-whale apomyoglobin on going from pH 8.0 to pH 6.0, cannot be ascribed to the formation of a charge transfer complex between a nearby histidine residue and W14 as reported in earlier papers but rather to minor structural changes affecting the microenvironments of both residues. The formation of the acidic partly folded state around pH 4.0 determines an increase of the fluorescence yield and a small red shift (5 nm) of W7 due to removal of sterically interacting K79, which is able to attenuate the emission of this residue in the native state. The fluorescence intensity of the other residue, i.e., W14, is not affected by the acidic transition. Guanidine denaturation experiments revealed an increase of fluorescence yield of W14 upon the intermediate formation, whereas the fluorescence of the other residue remained constant. The results suggest that the unfolding pathway may be different depending on the chemical nature of the denaturant used. PMID- 10669784 TI - Implications of the S-shaped domain in the quaternary structure of human arginase. AB - Arginase I is a homotrimeric protein with a binuclear manganese cluster. At the C terminus of each monomer, the polypeptide chain forms an unusual S-shaped oligomerization motif where the majority of intermonomer contacts are located [Z.F. Kanyo, L.R. Scolnick, D.E. Ash, D.W. Christianson, Nature 383 (1996) 554 557]. In order to study the implication of this motif in the quaternary structure of human arginase I, we have constructed a truncated arginase lacking the 14 C terminal amino acids, leaving Arg-308 as the last residue in the sequence. The resulting protein retains its trimeric structure, as determined by gel filtration (molecular mass 94 kDa). The same result was obtained in the presence of high ionic strength (KCl 0.5 M). Both data indicate that neither the S-shaped motif nor Arg-308 are fundamental in keeping the trimeric quaternary structure. Data obtained from intrinsic anisotropy and fluorescence intensity studies allow us to predict that the distance between the two unique tryptophans in the sequence is 2.9 nm in the native arginase and 4.1 nm for the truncated mutant. These distances allow us to assume a different conformational state in the truncated arginase without any change in its quaternary structure, suggesting that the carboxy-terminal motif is not the most prominent domain implicated in the quaternary structure of human arginase. Collisional quenching studies reinforce this possibility, since using I(-) as quenching molecule we were able to distinguish the two tryptophans in the truncated arginase. Moreover, kinetic studies show that the truncated mutant was fully active. In summary, the main conclusion about the structure of the human arginase I, derived from our study, is that the C-terminal S-shaped motif is not basic to the maintenance of the quaternary structure nor to the activity of the protein. PMID- 10669785 TI - Characterisation of endoglucanases EGB and EGC from Fibrobacter succinogenes. AB - The enzymatic properties of two endoglucanases from Fibrobacter succinogenes, EGB and EGC, were analysed. EGB and EGC were purified from recombinant Escherichia coli cultures expressing their gene. The failure of purification of EGB by classical techniques led us to produce antipeptide antibodies that allowed immunopurification of the protein from E. coli as well as its detection in F. succinogenes cultures. Synthetic peptides were selected from the predicted primary structure of EGB, linked to bovine serum albumin and used as immunogens to obtain specific antibodies. One of the polyclonal antipeptide antisera was used to purify EGB. EGC was purified by affinity chromatography with Ni-NTA resin. The endo mode of action of the two enzymes on carboxymethyl-cellulose was different. The values of K(m) and V(max) were respectively 13.6 mg/ml and 46 micromol/min mg protein for EGB, and 7 mg/ml and 110 micromol/min mg protein for EGC. The reactivity of the antipeptide and the anti-EGC sera with F. succinogenes proteins of molecular mass different from that of EGB and EGC produced in E. coli suggested post-translational modification of the two enzymes in F. succinogenes cultures. Expression of endB and endC genes in F. succinogenes was confirmed by RT-PCR. PMID- 10669786 TI - Engineering a compact non-native state of intestinal fatty acid-binding protein. AB - The last three C-terminal residues (129-131) of intestinal fatty acid-binding protein (IFABP) participate in four main-chain hydrogen bonds and two electrostatic interactions to sequentially distant backbone and side-chain atoms. To assess if these interactions are involved in the final adjustment of the tertiary structure during folding, we engineered an IFABP variant truncated at residue 128. An additional mutation, Trp-6-->Phe, was introduced to simplify the conformational analysis by optical methods. Although the changes were limited to a small region of the protein surface, they resulted in an IFABP with altered secondary and tertiary structure. Truncated IFABP retains some cooperativity, is monomeric, highly compact, and has the molecular dimensions and shape of the native protein. Our results indicated that residues 129-131 are part of a crucial conformational determinant in which several long-range interactions, essential for the acquisition of the native state, are established. This work suggests that carefully controlled truncation can populate equilibrium non-native states under physiological conditions. These non-native states hold a great promise as experimental models for protein folding. PMID- 10669787 TI - Inactivation of N-terminal signaling domain of Sonic hedgehog by forming a disulfide bond. AB - The N-terminal domain of mouse Sonic hedgehog (Shh-N) expressed in mammalian cells showed four-fold bands on non-reduced SDS-PAGE, though it was homogeneous under reduced conditions. It contains three cysteine residues, Cys-25, Cys-103, and Cys-184, which may be concerned with this heterogeneity. Therefore, we examined the formation of a disulfide bond in the recombinant Shh-N and identified three kinds of disulfides with a combination of peptide mapping and NH(2)-terminal amino acid sequencing analysis. Among them, one type of the Shh-N containing a disulfide bond of Cys-103/Cys-184 could be separated from the other Shh-Ns using reverse phase HPLC and had no activity of alkaline phosphatase induction in C3H10T1/2 cells. This molecule could also be made by denaturation of the purified Shh-N with guanidine-HCl under non-reduced conditions. On the other hand, the reduced Shh-N and the reduced S-methylated Shh-N at cysteine residues showed approximately 10-fold higher activity compared to the originally purified Shh-N. These results suggested that Shh-N was synthesized as an active form whose three cysteine residues did not form disulfide and inactivated finally by forming a disulfide bond between Cys-103 and Cys-184. PMID- 10669788 TI - Aggregation of an amyloidogenic fragment of human islet amyloid polypeptide. AB - Native human islet amyloid polypeptide (hIAPP) has been identified as the major component of amyloid plaques found in the pancreatic islets of Langerhans of persons affected by type 2 diabetes mellitus. Early studies of hIAPP determined that a segment of the molecule, amino acids 20-29, is responsible for its aggregation into amyloid fibrils. The present study demonstrates that the aggregation of hIAPP 20-29-Trp is a nucleation-dependent process, displaying a distinct lag time before the onset of rapid aggregation. Moreover, the lag time can be eliminated by seeding the sample of unaggregated peptide with preformed fibrils. In contrast to the expectation from the conventional model of nucleation dependent aggregation, however, the lag time of hIAPP aggregation does not depend on peptide concentration. To explain this observation, a modified version of the standard model of nucleation-dependent aggregation is presented in which the monomeric peptide concentration is buffered by an off-aggregation-pathway formation of peptide micelles. PMID- 10669789 TI - Escherichia coli cytosine deaminase: the kinetics and thermodynamics for binding of cytosine to the apoenzyme and the Zn(2+) holoenzyme are similar. AB - Recombinant Escherichia coli cytosine deaminase is purified as a mixture of Zn(2+) and Fe(2+) forms of the enzyme. Fe(2+) is removed readily by o phenanthroline to yield apoenzyme (apoCDase) that contains <0.2 mol of Zn(2+)per mol of subunit. ApoCDase was efficiently reconstituted to Zn(2+)CDase by treatment with ZnCl(2). The interaction of cytosine with apoCDase and Zn(2+)CDase was investigated at pH 7.5 and 25 degrees C by monitoring changes in intrinsic protein fluorescence. The values for the kinetic data K(1), k(2), and k(3) for Zn(2+)CDase were 0.25 mM, 80 s(-1), and 38 s(-1), respectively. The value for k( 2) was statistically indistinguishable from zero. The analogous values for K(1), k(2), and k(-2), (k(3)=0) for apoCDase were 0.157 mM, 186 s(-1) and approximately 0.8 s(-1), respectively. The overall dissociation constant of apoCDase for cytosine was 0.00069 mM, whereas the K(m) of Zn(2+)CDase for cytosine was 0.20 mM. The pre-steady state phase of the reaction was associated with an absorbance increase at 280 nm that was attributed to solvent perturbation of the spectrum of cytosine or enzyme. Formation of the Fe(2+)CDase-cytosine complex was too rapid to monitor by these techniques. PMID- 10669790 TI - Construction and characterization of a catalytic fusion protein system: P 450(11beta)-adrenodoxin reductase-adrenodoxin. AB - Cortisol is an important intermediate for the production of steroidal drugs and can only be synthesized chemically by rather complicated multi-step procedures. The most critical step is the 11beta-hydroxylation of 11-deoxycortisol, which is catalyzed by a mitochondrial enzyme, P-450(11beta). Various fusion constructs of P-450(11beta) with its electron transfer components, adrenodoxin and adrenodoxin reductase, were produced by cDNA manipulation and were successfully expressed in COS-1 cells from which the hydroxylation activities were assayed. It was demonstrated that the fusion protein required both adrenodoxin reductase and adrenodoxin for its activity and was not able to receive electrons from an external source. The fusion protein with all three components had less activity than P-450(11beta) alone, receiving electrons from coexpressed or internal electron transfer components. The activities of the fusion proteins were determined mainly by the fusion sequence. The fusion protein with a sequence of P 450(11beta)-adrenodoxin reductase-adrenodoxin was more active than that of P 450(11beta)-adrenodoxin-adrenodoxin reductase, 1.5- and 3-fold for bovine and human P-450(11beta), respectively. Modification of the linker region by extending the size of the linker with various peptide sequences in the bovine P-450(11beta) adrenodoxin reductase-adrenodoxin fusion protein indicated that the linker did not have significant effect on the P-450 activity. Taken together, the fusion protein obtained here can serve as a model for the investigation of electron transfer in P-450 systems and is of potential importance for biotechnological steroid production. PMID- 10669791 TI - Non-oxidative modification of lens crystallins by kynurenine: a novel post translational protein modification with possible relevance to ageing and cataract. AB - In humans, the crystallin proteins of the ocular lens become yellow-coloured and fluorescent with ageing. With the development of senile nuclear cataract, the crystallins become brown and additional fluorophores are formed. The mechanism underlying crystallin colouration is not known but may involve interaction with kynurenine-derived UV filter compounds. We have recently identified a sulphur linked glutathionyl-3-hydroxykynurenine glucoside adduct in the lens and speculated that kynurenine may also form adducts with GSH and possibly with nucleophilic amino acids of the crystallins (e.g. Cys). Here we show that kynurenine modifies calf lens crystallins non-oxidatively to yield coloured (365 nm absorbing), fluorescent (Ex 380 nm/Em 450-490 nm) protein adducts. Carboxymethylation and succinylation of crystallins inhibited kynurenine-mediated modification by approx. 90%, suggesting that Cys, Lys and possibly His residues may be involved. This was confirmed by showing that kynurenine formed adducts with GSH as well as with poly-His and poly-Lys. NMR studies revealed that the novel poly-Lys-kynurenine covalent linkage was via the epsilon-amino group of the Lys side chain and the betaC of the kynurenine side chain. Analysis of tryptic peptides of kynurenine-modified crystallins revealed that all of the coloured peptides contained either His, Cys or an internal Lys residue. We propose a novel mechanism of kynurenine-mediated crystallin modification which does not require UV light or oxidative conditions as catalysts. Rather, we suggest that the side chain of kynurenine-derived lens UV filters becomes deaminated to yield an alpha,beta-unsaturated carbonyl which is highly susceptible to attack by nucleophilic amino acid residues of the crystallins. The inability of the lens fibre cells to metabolise their constituent proteins results in the accumulation of coloured/fluorescent crystallins with age. PMID- 10669792 TI - The binding of Na(+) to apo-enolase permits the binding of substrate. AB - Enolase from rabbit muscle (betabeta-enolase) is inactivated by NaClO(4). Enolase free of divalent cations is more susceptible to inactivation by NaClO(4) than is enolase in the presence of Mg(2+). We find that substrate protects apo-enolase against inactivation, indicating that substrate can bind to enolase in the absence of a divalent cation. This binding is not due to contamination by trace levels of divalent cations since (1) it occurs even in the presence of EDTA or EGTA and (2) metal analysis by ICP (inductively coupled plasma) mass spectrometry did not reveal sufficient contamination to account for the protection. The binding of PGA to apo-enolase did require Na(+). When TMAClO(4) was used instead of NaClO(4), there was no protection by PGA. Protection was restored when TMAClO(4) plus NaCl were used. The inactivation of apo-enolase by NaClO(4) is due to dissociation into inactive monomers. We conclude that Na(+) binds to apo enolase, permitting substrate to then bind. Of the three known Me(2+) binding sites on enolase, we believe the most likely binding site for Na(+) is the carboxylate cluster of site 1, the highest affinity site of enolase. PMID- 10669793 TI - Novel allosteric effectors of the tryptophan synthase alpha(2)beta(2) complex identified by computer-assisted molecular modeling. AB - Tryptophan synthase is a pyridoxal 5'-phosphate-dependent alpha(2)beta(2) complex catalyzing the formation of L-tryptophan. The functional properties of one subunit are allosterically regulated by ligands of the other subunit. Molecules tailored for binding to the alpha-active site were designed using as a starting model the three-dimensional structure of the complex between the enzyme from Salmonella typhimurium and the substrate analog indole-3-propanol phosphate. On the basis of molecular dynamics simulations, indole-3-acetyl-X, where X is glycine, alanine, valine and aspartate, and a few other structurally related compounds were found to be good candidates for ligands of the alpha-subunit. The binding of the designed compounds to the alpha-active site was evaluated by measuring the inhibition of the alpha-reaction of the enzyme from Salmonella typhimurium. The inhibition constants were found to vary between 0.3 and 1.7 mM. These alpha-subunit ligands do not bind to the beta-subunit, as indicated by the absence of effects on the rate of the beta-reaction in the isolated beta(2) dimer. A small inhibitory effect on the activity of the alpha(2)beta(2) complex was caused by indole-3-acetyl-glycine and indole-3-acetyl-aspartate whereas a small stimulatory effect was caused by indole-3-acetamide. Furthermore, indole-3 acetyl-glycine, indole-3-acetyl-aspartate and indole-3-acetamide perturb the equilibrium of the catalytic intermediates formed at the beta-active site, stabilizing the alpha-aminoacrylate Schiff base. These results indicate that (i) indole-3-acetyl-glycine, indole-3-acetyl-aspartate and indole-3-acetamide bind to the alpha-subunit and act as allosteric effectors whereas indole-3-acetyl-valine and indole-3-acetyl-alanine only bind to the alpha-subunit, and (ii) the terminal phosphate present in the already known allosteric effectors of tryptophan synthase is not strictly required for the transmission of regulatory signals. PMID- 10669794 TI - Heat shock protein (hsp90) interacts with smooth muscle calponin and affects calponin-binding to actin. AB - Interaction of smooth muscle calponin with 90 kDa heat shock protein (hsp90) was analyzed by means of native gel electrophoresis and affinity chromatography. Under conditions used, calponin and hsp90 form a complex with an apparent dissociation constant in the micromolar range. The major hsp90-binding site is located in the N-terminal (residues 7-144) part of calponin. Addition of calponin to actin-tropomyosin complex results in formation of actin bundles. Hsp90 partially prevents bundle formation without affecting the molar ratio calponin/actin in single actin filaments or actin bundles. At low ionic strength, calponin induces polymerization of G-actin. Hsp90 decreases calponin-induced polymerization of G-actin. It is supposed that hsp90 may be involved in the assembly of actin filaments. PMID- 10669795 TI - Primary structure determinants of the pH- and temperature-dependent aggregation of thioredoxin. AB - Thioredoxins are small proteins found in all living organisms. We have previously reported that Chlamydomonas reinhardtii thioredoxin h exhibited differences both in its absorption spectrum and its aggregation properties compared to thioredoxin m. In this paper, we demonstrate, by site-directed mutagenesis, that the particularity of the absorption spectrum is linked to the presence of an additional tryptophan residue in the h isoform. The pH and temperature dependence of the aggregation of both thioredoxins has been investigated. Our results indicate that the aggregation of TRX is highly dependent on pH and that the differences between the two TRX isoforms are linked to distinct pH dependencies. We have also analyzed the pH and temperature dependence of 12 distinct variants of TRX engineered by site-directed mutagenesis. The results obtained indicate that the differences in the hydrophobic core of the two TRX isoforms do not account for the differences of aggregation. On the other hand, we show the importance of His-109 as well as the second active site cysteine, Cys-39 in the aggregation mechanism. PMID- 10669796 TI - Probing the interaction between N(1),N(4)-dibenzylputrescine and tRNA through (15)N NMR: biological implications. AB - NMR spectroscopy was used to characterize the binding properties of polyamines to Escherichia coli tRNA. The (15)N NMR spectra of three (15)N-enriched N substituted putrescine derivatives (DMP, DEP and DBP) were recorded in the presence of tRNA, and the spin relaxation times of the nitrogen nuclei were measured. From these data, the activation parameters for the rotational correlation times of the (15)N nuclei were determined. The present data indicate that the nature of the amino substituents does play a relevant role in controlling the polyamine-tRNA interaction. This study also provides a rationale for the in vivo antiproliferative effect of DBP against tumoral cells. PMID- 10669797 TI - MALDI-TOF mass spectrometry analysis of substrate specificity of lebetase, a direct-acting fibrinolytic metalloproteinase from Vipera lebetina snake venom. AB - Lebetase is a direct-acting fibrinolytic zinc metalloendopeptidase related in amino acid sequence to reprolysins which include both hemorrhagic and non hemorrhagic proteinases. Despite apparent structural similarities, fibrinolytic and hemorrhagic proteinases differ significantly in substrate specificity. In this study, we have examined the activity of lebetase I against biologically active peptides (bradykinin, kallidin, substance P) and 6-10 amino acid residues containing peptides synthesized according to cleavage regions of alpha(2) macroglobulin, pregnancy zone protein (PZP) and fibrinogen. Lebetase was found to have no activity against studied hexapeptides. Surprisingly, the best substrates for lebetase were substance P, and peptide fragment of PZP, both were cleaved at position Pro-Gln. Identification of the hydrolysis products of 15 peptides by MALDI-TOF mass spectrometry analysis indicates that lebetase possesses broad substrate specificity. The MALDI-TOF MS technique was proven to be highly efficient for the recovery and identification of the peptides released by lebetase hydrolysis. PMID- 10669798 TI - Anterograde axonal transport of Boc-Arg-Val-Arg-Arg-MCA hydrolyzing enzyme in rat sciatic nerves: cleavage occurs between basic residues. AB - Axonal transport of Boc-Arg-Val-Arg-Arg-MCA hydrolyzing enzyme activity was studied in rat sciatic nerves from 12 to 120 h after double ligations. The anterograde axonal transport increased and peaked 72 h after ligation. The optimum pH for Boc-Arg-Val-Arg-Arg-MCA hydrolyzing enzyme activity was 6.5 to 6.9 and did not require Ca(2+) for the activity. Two molecular forms with enzyme activity were identified by size-exclusion chromatography and the molecular masses of the two enzymes were estimated to be 98 and 52 kDa. Two enzyme activities were strongly inhibited by Hg(2+), Cu(2+) and trypsin inhibitors such as TLCK, antipain and leupeptin. It cleaved the substrate, Boc-Arg-Val-Arg-Arg MCA, between the dibasic sequence Arg-Arg, and needed a support of aminopeptidase B-like enzyme activity for the liberation of 7-amino-4-methylcoumarin. These results suggest that the enzyme is transported in rat sciatic nerves and involved in the post-translational processing of precursor proteins under the anterograde axonal transport. But there is absolutely no evidence for a role in precursor processing and such a putative role is purely speculative. PMID- 10669799 TI - Conformational studies of Actinobacillus actinomycetemcomitans leukotoxin: partial denaturation enhances toxicity. AB - A 114 kDa, water-soluble, cytotoxin secreted by the Gram-negative bacterium Actinobacillus actinomycetemcomitans (Aa) is similar in sequence to Escherichia coli alpha-hemolysin, but is non-hemolytic, killing leukocytes of select species, including humans. In this work, we investigated aspects of the water-soluble conformation of Aa toxin which relate to its biological, pore-forming activity. The toxin has five native tryptophans and fluorescence spectra were monitored in aqueous solutions in the presence of varying denaturants. Significant changes in the fluorescence spectra, without significant wavelength shifts, were induced by small additions of denaturants and changes in the temperature or pH. The fluorescence changes suggested that small perturbations in the aqueous environment resulted in structural changes in the toxin related not to a large unfolding but to more subtle conformational changes. Analytical ultracentrifugation showed the toxin to be a globular monomer in dilute aqueous solution. Circular dichroism spectroscopy showed about 25% alpha-helical structure which is largely maintained up to a temperature (65 degrees C) known to deactivate toxin activity. Changes in the cytotoxic properties of the toxin were monitored with flow cytometric analysis following preincubation of the toxin under mild conditions similar to those used in the fluorescence studies. These experiments showed that the pretreated toxin exhibited enhanced cell-killing potency on toxin-sensitive cells. The correlation of cytotoxicity with the changes in Trp fluorescence is consistent with the idea that partial unfolding of Aa toxin is an early, obligate step in toxin-induced cell kill. PMID- 10669800 TI - Rabbit phosphoglucose isomerase/neuroleukin/autocrine motility factor: cloning via interspecies identity. AB - Phosphoglucose isomerase is the first committed enzyme of glycolysis. The protein also has a variety of biological activities on mammalian cells. The molecular basis of these extracellular functions is unclear, and the high resolution three dimensional structure of a mammalian enzyme has not been described. We report here the cDNA and protein sequence for phosphoglucose isomerase from rabbit muscle. The sequence was obtained directly by PCR without the need to screen clones from a cDNA library and encoded active enzyme when expressed in bacterial cells. The 558 amino acid rabbit coding sequence is the same length as and highly similar (92% residue identity) to the sequences from human and pig and less so (88%) to the mouse enzyme. Non-conservative amino acid changes between the four mammalian sequences are concentrated in the first 35 and last five residues. The rabbit protein has an additional Cys residue and amino acid changes at five positions otherwise invariant in the mammalian enzymes. PMID- 10669801 TI - Characterization of the gene encoding the [Fe]-hydrogenase from Megasphaera elsdenii. AB - The gene encoding the [Fe]-hydrogenase from the anaerobic bacterium Megasphaera elsdenii has been cloned and sequenced. The gene is monocistronic, in keeping with the protein being a monomer. The translated protein sequence (484 residues, M(r)=53 kDa) comprises a small 2[4Fe-4S] ferredoxin-like domain and a large domain containing the catalytic site. Comparisons with other [Fe]-hydrogenase sequences, including two of which the crystal structures are known, show that the M. elsdenii protein is among the smallest of these enzymes and provide useful indications regarding the basic structural core common to all [Fe]-hydrogenases. It is, nevertheless, to be noted that the genome of Thermotoga maritima encodes a putative [Fe]-hydrogenase that would consist of only 301 residues. PMID- 10669802 TI - A common motif in proparts of Cnidarian toxins and nematocyst collagens and its putative role. AB - In Cnidarians, cnidoblast cells contain organelles called cnidocysts, which are believed to be the product of an extremely complex regulated secretory pathway. When matured, these stinging organelles are capable of storing and delivering toxins. We hypothesized that translated nematocyst proteins might comprise specific sequences serving as signals in sorting to the organelle. A sodium channel neurotoxin from the sea anemone Actinia equina was cloned and the toxin precursor sequence was compared to those of nematocyst collagens, pore-forming toxins and ion channel neurotoxins. It was found that all the analyzed sequences possess a highly conserved stretch of nine amino acid residues ending with Lys Arg N-terminally of the mature region. PMID- 10669803 TI - Cloning and nucleotide sequence of a gene encoding a glycogen debranching enzyme in the trehalose operon from Arthrobacter sp. Q36. AB - A gene located just upstream of the treYZ operon was isolated from Arthrobacter sp. strain Q36. The gene, designated treX, encoded an 823-amino acid protein. The amino acid sequence of the protein had 50% identity with the TreX protein (isoamylase) from Sulfolobus acidocaldarius ATCC 33909 which has a treZXY operon on the genome. We suggest that Arthrobacter treX is an isoamylase gene, and that it is a component of a treXYZ operon. PMID- 10669804 TI - Crystallization and preliminary X-ray crystallographic analysis of alginate lyase A1-II from Sphingomonas species A1. AB - Alginate lyase A1-II of Sphingomonas species A1 was purified and crystallized using the hanging drop vapor-diffusion method in 0.1 M Tris-HCl buffer containing 43% saturated ammonium sulfate, 8% polyethylene glycol 4000 and 0.2 M Li(2)SO(4) at pH 8.5 and 20 degrees C. The crystals are tetragonal and belong to the space group P4(3)2(1)2 or P4(1)2(1)2 with unit cell dimensions of a=b=144.07 and c=296.38 A. The diffraction data up to 2.8 A were collected by a synchrotron radiation source at SPring-8 in Japan. PMID- 10669805 TI - [Chemical weapons: antidotes. View about the real means, perspectives]. AB - Chemical methods remain a credible threat in 1999. The doctrine for their use not only includes the battlefield but also domestic terrorism as was disclosed during the Tokyo metro attempt in 1995. International Treaties have not yet proven their efficacy. The arsenal of chemical weapons has changed little since the second World War but is now dispersed into many high-risk zones throughout the world. There has also been little change in antidotes: therapeutic prevention with pyridostigmine against organo-phosphorus compounds, protective treatment for seizure-induced brain lesions using anticonvulsants in association with oxime for acetylcholinesterase reactivation, and atropine are combined in a three compartment syringe. Preventive measures against vesicants and other suffocating or toxic intracellular substances (CN, AsH(3), fluorocarbons.) can only be achieved with protective skin covering or protective breathing devices. There is no specific treatment and we often have to use symptomatic medications. Future perspectives include: phosphotriesterases as organo-phosphorus scavengers, huperzine as pretreatment and gacyclidine (GCK 11) which would effectively complete emergency multiple drug therapy against nerve agents. A new two compartment syringe is now prepared with atropine, avisafone and HI6 or pralidoxine. A gel made of cyclodextrines for external and eventually internal use is under study. PMID- 10669806 TI - [Pretreatment of organophosphate poisoning: potential interests of huperzine A]. AB - Pyridostigmine which is widely used as pretreatment of organophosphate poisoning protects cholinesterases of peripheral nervous system. Other molecules able to also protect the central nervous system are under study and, among them, huperzine A. This paper gives an overview of the current investigations about the efficacy and the innocuity of this molecule (study of the mechanisms of action, biological targets, behavioural manifestations) and brings out its potential interests. PMID- 10669807 TI - [Interest in an glutamate antagonistic compound for the treatment of organophosphate poisoning]. AB - Classical emergency treatment of organophosphate poisoning includes the combined administration of a cholinesterase reactivator (an oxime), a muscarinic cholinergic receptor antagonist (atropine) and a benzodiazepine anticonvulsant (diazepam). In subjects taking pyridostigmine and trained to autoadminister at least one autoinjector at the first signs of poisoning, classical emergency treatment ensures survival but only an antiglutamatergic compound like gacyclidine appears to be able to ensure optimal management of nerve agent poisoning in terms of rapid normalization of EEG activity, clinical recovery and total neuroprotection. All of this reinforces the therapeutical value of gacyclidine, which is in the process of approval for human use in France for treatment of head injury, as a central nervous system protective agent for the treatment of OP poisoning. PMID- 10669808 TI - [Substituted cyclodextrins as chelating reagents for ethers, thioethers and yperite]. AB - The complexation of mustard gas Cl(CH(2))(2)S(CH(2))(2) Cl, HD, yperite) and of ethers and thioethers derivatives by cyclodextrins: natural alpha-cyclodextrin (ACD) and substituted B-cyclodextrins was studied by NMR. A 1/1 stoechiometry was found in all cases, while affinity constants were found relatively weak (from 5 M(-1) to 100 M(-1)). However, these results show that chelation of HD by cyclodextrins can be reasonably expected, especially if chemical modifications provide stronger affinity constants. PMID- 10669809 TI - [The threat represented by toxins: myth or reality?]. AB - The reasons underlying our fear of the use of toxins as new weapons different from chemical agents are discussed together with the conditions required for their use on the battlefield. The undeniable scientific contribution of toxins as pharmacological tools for the study of neurosciences and disease genesis and their prophylaxis is emphasized. In conclusion, the threat for mankind constituted by proliferation of this novel class of agents is stressed. PMID- 10669810 TI - [Natural biological risks and military biological risks]. AB - The Iraqi biological program, the activities of sect Aum in Japan and the extensive endemicity of plague prove the existence of military, terrorist and natural biological risks. Among the agents of natural risk (viruses, bacteria.), plague is induced by modification of the ecosystem. Present since 1921 in the high plateau of Madagaskar, the disease evolves under two modes, endemic (natural) or epidemic (urban). Since the control of endemicity is impossible, the decrease of incidence will be obtained by the control of the animal reservoir. The military risk is part of the history of armed conquests. Anthrax and botulinum toxins, are the most toxic agents, banned by the Convention of London (1972). In 1995, 4 years after the end of Gulf war, UNSCOM obtained from authorities the inventory of Iraqi biological program, with details on the militarization of toxins and spores. These furtive weapons, are produced with limited technological skills, often in dual manufactures and are difficult to control. PMID- 10669811 TI - [Risks related to the uncontrolled development of pharmacognosy in the United States and France]. AB - A number of plants of ancestral tradition exhibit properties deserving therapeutic applications. However, their use in conditions which do not guarantee sufficient safety may lead to incidences and sometimes serious accidents. In the US the weakness of the present provisions of the federal regulation do not allow a satisfactory and preliminary control. Moreover, these plants are mostly prescribed by non-medicals and outside the authority of pharmacists. This situation would be highly prejudicial if it would be implemented in Europe. France has a well conceived and satisfactory system, but which is unable to benefit from new accessions, namely exotic plants. To ignore them would lead to strengthening the already existing and parallel organizations. The authors suggest that the health authorities should create a complementary system open to new medicinal plants, avoiding uncontrolled sales. They suggest also that before any acceptance a realistic probationary period would be applied in order to evaluate the quality but overall the eventual toxicity of these plants. PMID- 10669812 TI - [Pharmaceutical development concerning diseases predominating in tropical regions: the concept of indigent drugs]. AB - When the WHO certified the eradication of smallpox in 1981, there was a general impression that the fight against infectious diseases which began with Jenner and Pasteur was entering a phase of achievement: poliomyelitis, dracunculasis, leprosy, Chagas' disease and neonatal tetanus were also responding to eradication campaigns. However, in 1995, infectious diseases are still an important cause of mortality and morbidity and the rising incidence of emerging or re-emerging diseases remains a matter of great concern. Although this situation can be explained, at least partly, by the deterioration of health care systems and diverse socio-economic and ecological disorders, important changes occurring in the drug industry since 1980 have also played a role due to changes in pharmaco epidemiology and new policies of drug development. Among the 1061 new drugs developed from 1975 to 1994, less than 2.7% concern tropical diseases. Since praziquantel, novel drugs have issued from veterinary medicine (ivermectin), military research (halofantrine, mefloquine) or fortuitous analysis of pharmacopoeia (artesunate). The cost of investments and the lack of market potential and market security in developing countries have dampened interest in developing drugs for tropical diseases. Observing the combined effect of deficient pharmaceutical development, drug wear due to chemoresistance (chloroquine, sulfadoxine-pyrimethamine, aminopenicillins), the cost barrier (second generation molecules) and the potential abandon of major drugs (eflornithine, melarsoprol) has led us to establish a classification of these "indigent" drugs (in opposition to "orphan" drugs) into five classes: true indigent drugs (eflornithine), indigent drugs by indication (pentamidine), indigent drugs by function (ceftriaxone), indigent drugs by formulation (melarsoprol) and indigent drugs by default (suramin). This analysis can serve as a basis for a search for solutions (regulatory, administrative and financial incentives) favoring a reactivation of drug development for diseases predominating in intertropical regions. PMID- 10669813 TI - Interactions between chitosan and glycosaminoglycans (chondroitin sulfate and hyaluronic acid): physicochemical and biological studies. AB - Chitosan is a polysaccharide well known for its numerous and interesting biological properties, indeed, it is a biocompatible, bioresorbable and bioactive biopolymer. It is therefore often introduced in the human body, where it happens to be in contact with glycosaminoglycans (GAG's), especially chondroitin sulfates and hyaluronic acid which play important part in living media. Thus it seems interesting to consider systems associating chitosan and GAG's. The aim of our work is to study the mechanism of formation of biomaterials constituted of chitosan and GAG's which could have interesting biological properties such as improving the wound-healing acceleration and the cellular assistance for skin and cartilage recovery. In the same time, it was interesting to try to understand what can happen when chitosan is introduced in a natural surrounding containing GAG's. In a first part, the complexes between chitosan and GAG's were characterised by various physicochemical techniques: pH-metry, conductometry and Infra-red spectrometry. They appeared to be polyelectrolyte complexes obtained by the formation of polyanion-polycation interactions held between ammonium functions of chitosan and carboxylate and/or sulfate groups of GAG's. The complexes formed are very strong and can, in some cases, even conduct to the deprotonation of carboxylic functions. In a second part we considered the biological properties of the complexes previously obtained. The hydrolysis activity in presence of specific enzymes was investigated. We also studied the cytocompatibility of chitosan alone and its complexes with the two considered GAG's toward two kinds of cells (chondrocytes and keratinocytes). At the end of the work, a short animal experimentation on rats was performed allowing the comparison between in vitro and in vivo results. PMID- 10669814 TI - [Transgenic mice expressing MTCP1: an animal model for T-cell prolymphocytic leukemia]. AB - T-prolymphocytic leukemia (T-PLL) is a rare form of mature T-cell leukemia associated with chromosomal rearrangements of band 14q11, containing the gene TCRA/D, and bands 14q32.1 and Xq28, where the TCL1 and MTCP1 putative oncogenes have been identified. These genes encode two homologous proteins, p14(TCL1) and p13(MTCP1) respectively, which share no similarity with other known proteins. To determine the oncogenic role of MTCP1, transgenic mice with an expression of MTCP1 targetted to the T-cells were generated. A lymphoid malignancy similar to Human T-PLL occurred in to independent transgenic lines with a high level of expression of the transgene. The cumulative incidence of the disease at 20 months was 100% and 50% respectively, and null in the control group. The oncogenic role of MTCP1 is demonstrated, and the p13(MTCP1) and p14(TCL1) proteins form a new oncoprotein family. The long latency period before emergence of tumors suggests that activation of MTCP1 is not sufficient to generate the malignant transformation. The secondary genetic events implicated in tumoral progression remain to be elucidated, in order to reconstruct the molecular history of the disease. PMID- 10669816 TI - [ [In Process Citation] PMID- 10669817 TI - [ [In Process Citation] PMID- 10669815 TI - [The prescription in dispensary practice]. AB - Although the medical prescription is a well-established process, prescribing in the pharmacy does not follow a well-defined set of rules as, in official terms, there is no such thing as a pharmaceutical prescription. Despite this fact, daily pharmacy practice involves giving professional advice and dispensing drugs not only in fulfillment of a medical practitioner's prescription but also by selling drugs the pharmacist personally advises use of. In this particular case, the pharmacist must assess the patients desire for a specific treatment and determine whether it is well founded. The pharmacist has to decide whether he/she can deal with the presenting symptom or whether the person should be referred to a practising physician. For example, often-recurring mouth disease such as mouth ulcers and gingivitis can be handled within the pharmacy. The pharmacist may think of implementing a suitable treatment involving dietary and hygienic counselling and dispense carefully selected drugs. The situation is similar in many other clinical conditions. As the pharmacist has had professional training in drugs at the university and teaching hospital, he/she is the expert in drugs and can rightly claim entitled to prescribing drugs within reasonable limits. PMID- 10669818 TI - [2000-2010: bone and joint decade]. PMID- 10669819 TI - [Radiographic assessment of polyethylene wear in machined versus molded polyethylene in total hip prosthesis]. AB - PURPOSE OF THE STUDY: Wear of polyethylene acetabular components is an important issue in total hip arthroplasty. This study was designed to evaluate differences in polyethylene wear rates between machined and direct compression molded acetabular cups. METHODS: Two hundred thirty-nine prostheses underwent radiographic evaluation using the technique of Chevrot-Kerboull. One hundred thirty-one were all-polyethylene cups machined from extruded bar stock, and one hundred eight were all-polyethylene cups direct compression molded. Both groups, all operated on in 1988, were similar in the acetabular and femoral components, were all cemented, and the acetabular components were all-polyethylene, non metal backed. The femoral components were all Charnley-Kerboull MK III type. Patient weight, average weight, the post-operative PMA score and the duration of the follow-up were similar for the two groups (mean 7.4 years). RESULTS: Results showed a mean linear wear rate of 0.06 mm per year for compression-molded polyethylene and 0.08 mm per year for machined polyethylene. Results were not significantly different. The number of acetabular radiolucencies in zone I (5.5 versus 5.3 p. 100) and the amount of lysis of the proximal part of the femur did not differ between the two groups. The number of excessive wears did not differ either. DISCUSSION AND CONCLUSION: Although polyethylene wear was lower with compression molded acetabular cups than with ram-extruded acetabular cups, results of this study suggest a non significant difference between the two types of components. A more controlled experiment would have to be performed to attribute a difference between two different types of UHMPE processing. PMID- 10669820 TI - [Oxidation of ultra high molecular weight polyethylene as a result of sterilization]. AB - PURPOSE OF THE STUDY: To improve (UHMWPE) Ultra High Molecular Weight Polyethylene quality for use in arthroplastic components, sterilization related oxidative changes were investigated in raw and manufactured material. MATERIAL: To evaluate sterilization related effects, 15 x 15 mm samples of UHMWPE were taken from a defined area of raw manufactured UHMWPE plates. The raw manufactured plates were produced using the compression molding method. For sterilization, gamma irradiation with and without air, ETO-sterilization and autoclave sterilization were performed. METHODS: Infrared spectroscopy was used to detect oxidation damage of the implant in superficial and deep layers of the material. RESULTS: Gamma sterilization on air showed 40% more oxidation compared with air free sterilization. The superficial oxidation on air was 0.33 and showed the lowest value without air on argon with 0.155. Using ethylenoxide sterilization, no great changes in superficial oxidation (0.229) were observed; the smallest change (0.07) was observed at a depth of 2.79 mm. Autoclave sterilization caused the greatest superficial oxidation with 0.622 but low oxidation at 0.094 mm depth. In all samples, further oxidation and ongoing crystallinity was seen with increasing storage time. CONCLUSION: Ethylenoxide sterilization should be recommended for UHMWPE since oxidative changes is lowest. Gamma sterilization can only be recommended without air and at low doses. PMID- 10669821 TI - [Osteolysis after total knee prosthesis]. AB - PURPOSE OF THE STUDY: The aim of our work was to study X-rays showing osteolysis after 5 years and more in 122 prosthesis and to try and assess such complication, often described in the United States but seldom in Europ. MATERIAL: We are dealing here with 122 retaining posterior cruciate ligament, mostly cementless prothesis implanted between 1985 and 1992 84 chromium-cobalt prosthesis (PCA and Themis) implanted in 34 males and 88 females with an average age of 67 (45-81), 87,7 p. 100 had femoral cementless components and 70 p. 100 tibial cementless components. METHODS: All patients were examined and had X-rays at an average of 6,9 years. Specially considered were X-rays showing a possible osteolysis. We looked for possible complication (external laxity, anterior femoral dislocation and polyethylene wear), assessment of the mechanical axis and for clinical results (Hungerford score) RESULTS: Revisions: 19 arthroplasties were revised for PE wear tibial loosening metallosis or patella problems The postoperative score according to Hungerford was 84,5 p. 100 for PCA prosthese and 87 p.100 for the Themis. On the X-rays were only few osteolysis to be found: 9 cases (7,3 p. 100). For the PCA series: 3 femoral osteolysis, 1 tibial at 12 years, and one patellar osteolysis. For the Themis series: no femoral osteolysis, 3 tibial and one patellar osteolysis. Osteolysis are apparent on X-rays in profile for the femur and the patella, and in both profile and frontal X-rays for the tibia. Clinicaly 4 osteolysis really asymptomatic were not re-operated. 5 were revised: one 11 years later for femoral and tibial loosening, two for a patellar loosening, the other two patients had to be reoperated on for metallism (titanium's femoral component) and for those two instances osteolysis were discovered during the complication. DISCUSSION: Osteolysis after TKA appears unusual in our experience without bearing on frequency finded by american authors with a lesser follow-up (Engh 11,1 p. 100 after 4,5y, Peters 16 p. 100 after 2,9 y, Robinson 9,18 p. 100 after 4,6 y). American litteratur analysis shows that the important number of osteolysis is due to: - either to dual-metal using (Co-Cr component with Titanium screws for exemple), - or bad quality of polyethylene (compressed), - or a bad design of former prosthesis. CONCLUSION: Interface illness, linked to the production of wear debris, osteolysis after total knee arthroplasty is rarer than after a hip one, probably because size of debris is different, larger in knee than in hip. It is likely that the improvement of PE quality, design of prosthesis, as well as a better knowledge of osteolysis mechanism will allow to delay this complication wich is in a long term ineluctable. PMID- 10669822 TI - [Tibial fracture with intact fibula treated by reamed nailing]. AB - PURPOSE OF THE STUDY: The main difficulties encountered in the orthopedic treatment of leg fractures with intact fibula are reduction of the tibial and an unusually high rate of varus unions and non-unions. The aim of this retrospective study was to assess the outcome after reamed nailing of tibial fractures with an intact fibula. MATERIAL AND METHOD: Between 1986 and 1997, 38 fractures of the tibia with an intact fibula were treated by first intention centromedullar nailing. There were 28 men and 10 women, mean age 28 years, with a single fracture in 25 cases. There were 25 motor vehicle accidents (17 two-wheel, 8 four wheel), 5 sports accidents, 2 home falls, and 6 others. Fracture of the tibial diaphysis was associated with a homolateral femoral fracture in 7 cases, 7 fractures were open (7 type 1, 2 type 2, 1 type 3), 7 fractures were associated with abrasive skin lesions. Using the AO classification, the tibial fracture was type A in 26 cases, type B in 11, and type C in 1. The fracture was in the middle third of the tibia in 21 cases, the distal third in 15 and in the proximal third. Grosse and Kempf nails were used exclusively. Static nailing was used in 27 cases, dynamic nailing in 8, and the nail was not locked in 3 cases. Nails of diameter 9 to 13 were implanted after reaming 1 mm more. RESULTS: The fracture gap increased during the reaming in 5 patients; 2 patients had to undergo a secondary aponeurectomy due to a postoperative compartment syndrome and had no further sequela. Consolidation was achieved after the first intention treatment in 30 patients, after dynamization in 6. A non-union in 2 patients was also successfully managed with new nailing and dynamization. Delay to consolidation was a mean 175 days (range 60 - 480). Transverse fractures consolidated more rapidly (mean 122 days). At last follow-up (minimum 1 year), active knee and ankle mobility were normal in all patients. Nineteen patients complained of pain at the site of the nail insertion, evaluated at 1 on a 10-point analogie scale by 10 of them and at 2 by the 9 others. Eight out of 10 patients felt cure had been achieved 5 months postoperatively. DISCUSSION: These rapidly obtained clinical results and the relatively low rate of non-union (5 p. 100) should be attributed to the reamed nailing technique. We discuss the frequency of tibial fractures with intact fibula and the underlying circumstances. The lack of patent fibular fracture does not signify the fibula is intact. Trauma-induced tibio-fibular dislocation (1 case in our series) can occur. A review of the literature emphasizes the frequency of non unions and misalignment after orthopedic treatment. The most widely used surgical technique is reamed nailing. This technique has the inconvenience of possible pain at the insertion site which usually disappears after ablation of the nail and also a compartment syndrome where reaming is a possible aggravating factor. CONCLUSION: Nailing is a reliable technique for the treatment of tibial fractures with an intact fibula. Weight bearing should be encouraged as early as possible. The indication for a locked nail depends on the anatomic type of the tibial fracture and its localization. Immediate weight bearing should be recommended. Strict surveillance allows dynamization with fibulotomy in case of late consolidation. Prospective randomized studies comparing nailing with other therapeutic methods are needed to confirm these data. PMID- 10669823 TI - [Subcutaneous tenotomy of Achille's tendon in adults for ankle stiffness. A review of 80 cases]. AB - PURPOSE OF THE STUDY: Lengthening of Achille's tendon is part of surgical program for ankle stiffness in equinus deformity. Usually this lengthening is done by opened surgery with all the well-knowned advantages in term of adjustment. We used a percutaneous method for Achille lengthening with a two stages tenotomy. The purpose of this study is to evaluate advantages, inconvenients and efficiency of the subcutaneous method versus the open method. MATERIALS AND METHODS: We present a retrospective study of a 80 percutaneous lengthening of Achille's tendon in 78 patients done between August 85 and January 96. All patients who went to surgery during this period were reviewed. Mean age was 36 years old and there was 39 left sides and 41 right sides. We separated the extra-articular stiffness (48 cases) and the intra-articular stiffness (32 cases) because in extra-articular etiology the stiffness is a consequence of a primitive neuro muscular disease as the ankle joint is healthy. 46 ankles had a past history of surgery. The kind of surgery was directely related to the etiology of the equinus. Most of the time, Achille lengthening was the last time of joint mobilisation. Only 15 times lengthening of the equinus tendon was done isolated. We looked at the early results for all patients and late results were only evaluated for the patients who had an isolated lengthening of the Achille tendon the for stastitical reasons. RESULTS: Median follow-up is two and an half years. There was preoperatively 59 ankles equinus and correction was constantly obtained with surgery postoperative improvment was 12 degrees of dorsal ankle flexion. In sub population of isolated achillus lengthening mean gain is 17 degrees. Only one patient had a per-operative complication with an heel anesthesia. We had no late complication related to the method. DISCUSSION: Results of this reviewal confirms efficency of the percutaneous technique in adult. Litterature is very poor concerning Achille's tendon lengthening in adult surgery. It is a very widelly spread method in children. Only few authors have published about it in adults mainly about hemiplegic patients. Our method saves tourniquet time in heavy ankle surgery. There are no painful and sticky scar. Morbidity is very low because we only had one complication related to the method. Healing up of tendon is very good, all patients being able to rase up on their toes. CONCLUSION: A review of our cases showed us the real efficency of this technique. It's a very simple and quick method giving good results with low morbidity. For us there is no indication of open surgery for achille's tendon lengthening. PMID- 10669824 TI - [Neurogenic mixed evoked potential monitoring during scoliosis surgery: retrospective analysis of 149 cases]. AB - INTRODUCTION: We report a retrospective analysis of spinal cord monitoring with neurogenic mixed evoked potentials (NMEPs) combined with somatosensory evoked potentials (SSEPs) in 149 patients undergoing surgery for spinal deformity. MATERIAL AND METHODS: 149 patients (104 females and 45 males), mean age 28 yrs (13-72 yrs) were studied. NMEPs were elicited by electrical spinal cord stimulation in the rostral part of the surgical field, via two needle electrodes set in the epidural space and in the interspinous ligament above. They were recorded from the sciatic nerve at the knee and the sural nerve at the ankle. SSEPs were recorded from the scalp after stimulation of the posterior tibial nerve at the ankle. A decrease in amplitude of more than 50 p. 100 and/or an increase in latency of more than 10 p. 100 were defined as significant warning criteria. RESULTS: No false-negative result was observed. NMEP modifications did not reach critical value in 143 cases. In 6 cases, significant changes were observed. Moving the stimulation electrodes along the spinal cord allowed spinal lesion localization and helped the surgeon to perform the adapted maneuver, clearly avoiding the occurrence of postoperative neurological defect in 5 of the 6 cases. CONCLUSION: NMEP monitoring is a sensitive and specific method useful for detecting an impending lesion of the spinal cord. NMEPs are also helpful in localizing the spinal level of the lesion. They represent a primary choice tool for neuromonitoring during scoliosis surgery. PMID- 10669825 TI - [Surgical treatment of hallux valgus in children and adolescents: 46 cases treated with the Mitchell technique]. AB - PURPOSE OF THE STUDY: The aim of this study is to discuss special features of hallux valgus in children and assess place of a surgical procedure in some patients. MATERIAL AND METHOD: A retrospective study of 40 children (46 feet) operated for hallux valgus was conducted. An etiology was identified in 10 per cent of cases and 90 per cent were considered as idiopathic. From a clinical and radiological analysis of 46 feet, we describe the anatomical particularities and the indications of the surgical treatment. The Mitchell's technique is described in detail. The correction of the metatarsal varus and the metatarsophalangial valgus was obtained without breaking off the growth plate of the first metatarsus. RESULTS: Clinically, results were satisfactory in 84 per cent of the cases. Radiologically, results were excellents in 45 per cent and good in 35 per cent. DISCUSSION AND CONCLUSION: Hallux valgus in the child is often underestimated; operative treatment should be proposed in some patients. Mitchell's technique gives good results with an early and simply correction. PMID- 10669826 TI - [Primary leiomyosarcoma of bone. Report of 5 anatomo-clinical cases and review of the literature]. AB - INTRODUCTION: Leiomyosarcoma is a malignant smooth muscle tumor occurring most frequently in uterus or soft tissues and more rarely in bone. MATERIALS AND METHODS: We report the clinicopathologic, immunohistochemical and ultrastructural findings of five cases of primary leiomyosarcoma of bone treated in our Department between 1991 and 1994. The pertinent medical literature is discussed. RESULTS: The tumors were located respectively in the distal tibia (n=2), the distal femur, the sternum and the ilium (n=1). Four lesions were high-grade and one low-grade. All patients (3 women and 2 men) underwent wide surgical resection associated with polychemotherapy in four cases. Two patients died of metastatic disease, two had local recurrence and one is alive with no evidence of disease at the last follow-up. DISCUSSION: Excluding cases which involve the facial skeleton, there are to our knowledge 95 cases of primary leiomyosarcoma of bone reported in the literature. This tumor arises more commonly in adults (mean age: 49 years) with an equal gender distribution and involves predominantly the long bones near the knee. In the majority of cases, plain X-rays exhibit an osteolytic lesion with cortical penetration and indistinct margins. The diagnosis is based on microscopic features demonstrating fusiform tumor cells arranged in interwoven bundles, and the immunohistochemical results of widespread cytoplasmic positivity for smooth muscle actin. The best pronostic parameter is the histologic grade correlated with both the recurrence and metastatic rates as well as the survival rate. Surgery constitutes the main treatment since chemotherapy or radiotherapy did not provide an improved prognosis over a wide resection. PMID- 10669827 TI - [Pseudotumoral subacute osteomyelitis: a series of 41 children]. AB - PURPOSE OF THE STUDY: To confirm that subacute osteomyelitis in children is a real entity, and give the main characteristics of this disease MATERIAL AND METHODS: We reviewed 41 cases as well as the litterature. RESULTS: Pain without fever was the only constant symptom. The erythrocyte sedimentation rate was increased slightly. XRays were the most important investigation, showing a geode in the metaphysis or more rarely in the epiphysis evoking a benign bone tumor in a child. DISCUSSION: The treatment was most often surgical: scraping permits the isolation of the microbe in 50 per cent of cases (staphylococcus most often) and usualy leady to recovery. CONCLUSION: This form of osteomyelitis is relatively frequent and must be discussed in front of a cystic tumoral image of the child's long bone. PMID- 10669828 TI - [Primary carpal bone defect]. AB - We present a review of management options in case of carpal bone defect, a relatively frequent discovery. In the literature, diagnosis is usually a fortuitous radiographic finding showing one or several images of carpal defect. Pain is observed in some cases, more exceptionally pathological fracture. The scaphoid, lunatum and hamatum are most frequently involved. Bilateral defects may be observed. Different mechanisms have been put forward to explain the development of intraosseous defects in the carpal bones including intraosseous penetration of synovial tissue, or in situ metaplasia of bone tissue. The main differential diagnoses are osteonecrosis sequellae (for the lunatum and the scaphoid), subchondral defects due to hyperpression and arthropathies in dialysis patients. All authors propose simple surveillance for asymptomatic images. In case of pain, with soft tissue swelling or pathological fractures, filling excision is warranted depending on the severity of the clinical signs. Prognosis is generally good and recurrence exceptional. PMID- 10669829 TI - [Extravasation injuries and subcutaneous aspiration in children]. AB - Complications of extravasation of intravenous fluid are not rare in pediatric practice, and their local (skin necrosis) and regional (vasculo nervous) implications can be severe. Two cases are presented in order to discuss the different treatments options and to describe the subcutaneous aspiration technique, which can be immediately efficient if proposed in a short delay. PMID- 10669830 TI - [Double dislocation of the fifth metacarpal. A case report and review of the literature]. AB - A case of traumatic double dislocation of the fifth metacarpal is reported. Both dislocations, hamatometacarpal and metacarpophalangeal, were dorsal. This rare combination of injury has been reported twice only. Our case was managed successfully by closed reduction and immobilization. Eight month later, the patient had a full range of wrist and finger movement, he was pain-free without any residual disability. PMID- 10669831 TI - [Aggressive osteoblastoma of the carpal scaphoid bone]. AB - We report a case of aggressive osteoblastoma of the scaphoid bone and discuss the features of this tumor. The few reported cases emphazise the rarity of osteoblastoma in hand and wrist localizations. Pathological examination is mandatory before treatment due to lack of distinctive clinical and radiological features. In our case, the rapidiy aggressive progression, similar to a malignant tumor, required radical treatment with scaphoidectomy. PMID- 10669832 TI - [Elastofibroma in the scapular region. A case report and review of the literature]. AB - PURPOSE OF THE STUDY: We report a case of elastofibroma and have collected 280 cases in the literature. MATERIAL AND METHODS: A 56 year-old man presented with a right subscapular mass. The patient was asymptomatic but he reported a "clicking"sensation associated with mobilization of the shoulder. Physical examination revealed a round mass clearly demonstrated with forward elevation of the arm. The MRI scan showed a heterogeneous soft tissue composed of inhomogeneous density with areas of more intense signal suggesting adipose tissue. The tumor was surgically excised and the diagnosis of elastofibroma was established by histopathologic examination. RESULTS: Six months after removal of the mass, there were no functional complications. DISCUSSION: Elastofibromas usually occur in active patients generally older than 55 years of age. They are typically located in the right subscapular region. The tumor remains asymptomatic in more than 50 percent of cases. 25 percent of the patients may report a simple discomfort sometimes with a "clicking" or "catching" sensation associated with mobilization of the arm. Pain is present in less than 10 percent of cases. Physical examination may reveal a rubbery, asymptomatic mass located in the subscapular region and barely noticable when the arm lies again the chest. Plain radiographs and preoperative laboratory data were unremarkable. CT scan or MRI scan may show an heterogeneous fibrous mass of intermediate density with entrapped signals of higher intensity. However, a definitive diagnosis requires a biopsy showing the distinctive feature of elastofibroma: elastic fibers in a collagenized fibrous tissue with entrapped adipose tissue. Pathogenesis of elastofibromas may result from the friction of the scapula against the thorax thus generating tumor growth. CONCLUSION: Complete surgical excision in symptomatic patients is considered to be the treatment of choice. However, once the diagnosis of elastofibroma has been established, excision of lesions smaller than 5 cm can be avoided in asymptomatic patients. PMID- 10669833 TI - [Re: "Cadaver study of acetabular cup mobility in the healthy hip and hip prosthesis by monopodal pressure simulation"]. PMID- 10669835 TI - [Pierre teinturier] [In Process Citation] PMID- 10669834 TI - [Re: "Free musculo-tendinous flap stabilization-interposition of the palmaris longus tendon after trapezium surgery in rhizoarthrosis]. PMID- 10669837 TI - Epithelial-mesenchymal interactions in the pathogenesis of asthma. AB - During lung development, repair, and inflammation, local production of cytokines (eg, transforming growth factor-beta) and growth factors (eg, epidermal growth factor) by epithelial and mesenchymal cells mediate bidirectional growth control effectively creating an epithelial-mesenchymal trophic unit. In asthma the bronchial epithelium is highly abnormal, with structural changes involving separation of columnar cells from their basal attachments and functional changes including increased expression and release of proinflammatory cytokines, growth factors, and mediator-generating enzymes. Beneath this damaged structure there is an increase in the number of subepithelial myofibroblasts that deposit interstitial collagens causing thickening and increased density of the subepithelial basement membrane. Our recent studies suggest that the extent of epithelial damage in asthma may be the result of impaired epidermal growth factor receptor-mediated repair. In view of the close spatial relationship between the damaged epithelium and the underlying myofibroblasts, we propose that impaired epithelial repair cooperates with the T(H)2 environment to shift the set point for communication within the trophic unit. This leads to myofibroblast activation, excessive matrix deposition, and production of mediators that propagate and amplify the remodeling responses throughout the airway wall. PMID- 10669838 TI - Parasites and asthma/allergy: what is the relationship? AB - Asthma prevalence is increasing in Western industrialized countries. The infectious theory of asthma onset hypothesizes that lower levels of IL-12 result in reduced T(H)1 stimulation and failure of the neonate to deviate from its T(H)2 bias at birth. Helminthic infections may influence T(H)2 immune responses and hence immune development. Although ecologic data would support a protective effect of parasitic infection on asthma development, this may be due to other exposures. To date, there is no conclusive evidence that parasitic infection protects against asthma development. PMID- 10669840 TI - Urbanization and childhood asthma: an African perspective. AB - The increasing prevalence of childhood asthma in the developed world is a cause for concern. Much research is currently being conducted in an attempt to identify possible reasons for this occurrence. A so-called Western lifestyle has been the factor most commonly cited to explain this worrying increase in asthma prevalence. In essence, this implies a way of life where children are exposed from early infancy to a wide range of foods, infections, indoor and outdoor allergens, and irritants and to the effects of motor vehicle pollution. Until fairly recently, children in many African countries lived mainly in rural areas and were not exposed to the effects of a Western lifestyle. Early studies in a limited number of African countries showed a very low rural prevalence of childhood asthma, especially where children lived according to a traditional lifestyle. These same studies showed that asthma was not uncommon in urbanized African children. There has been an increasing tendency over the past 20 years for those in rural communities to move to the large urban centers. More recent childhood asthma prevalence studies, especially those from Kenya and Ghana, have confirmed the urban-rural differences but have shown a much narrower gap. In part this may be the result of exposure of rural children to agricultural pesticides and irritants as well as of an increasing tendency to adopt a more Westernized lifestyle such as the use of beds with mattresses, pillows, and blankets. These circumstances on the African continent provide a natural laboratory in the quest for factors that influence the development of asthma in susceptible children. Once more fully elucidated, it is possible that much valuable information will be available to combat the relentless increase in childhood asthma both here as well as in the developed world. PMID- 10669839 TI - Molecular pathology of allergic disease. II: Upper airway disease. AB - Allergic upper airway diseases such as allergic rhinitis and chronic sinusitis are an increasing problem. Although the pathogenesis remains elusive, an individual's genetic predisposition as well as exposure to the allergen are currently considered factors in their development. Clinical symptoms of sneezing, rhinorrhea, and congestion are primarily a consequence of granulocyte release of chemical mediators such as histamine, prostanoids, and leukotrienes as well as the infiltration of inflammatory cells. Observations subsequent to allergen provocation are comparable to natural exposure and as such much of our understanding of allergic responses is derived from this model. A prominence of CD4(+) T cells and eosinophils, synthesis and release of T(H)2 cytokines, and the coordinate expression of chemokines and adhesion molecules are all characteristic of the allergic response observed in rhinitis and sinusitis. Corticosteroids and immunotherapy target these inflammatory processes and have been observed to successfully reduce and shift the predominantly T(H)2 environment toward T(H)1 cytokine expression. As our understanding of the pathophysiologic features of allergic upper airway disease improves, as well as the relationship between their development and that of lower airway disease, new strategies of diagnosis and treatment will allow for more effective modulation of the allergic process and associated morbidity. PMID- 10669841 TI - IL-11 expression is increased in severe asthma: association with epithelial cells and eosinophils. AB - BACKGROUND: IL-11 is a pleiotropic cytokine produced by a variety of stromal cells. Targeted overexpression of this cytokine in mice results in a remodeling of the airways and the development of airway hyperresponsiveness and airway obstruction. OBJECTIVES: Because these alterations mimic important pathologic and physiologic changes in the airways of some asthmatic patients, we investigated the expression of IL-11 messenger RNA (mRNA) within the airways of patients with mild to severe asthma and nonasthmatic control subjects. METHODS: Fiberoptic bronchoscopy to obtain bronchial biopsy specimens was performed on patients with mild (n = 13), moderate (n = 10), and severe (n = 9) asthma and on nonasthmatic control subjects (n = 9). RESULTS: These patients differed in their extent of airway fibrosis with types I and III collagens being noted in greater quantities in the biopsy specimens from the severe and moderate asthmatics than in those from controls (P <.05). IL-11 mRNA expression was observed in the epithelial and subepithelial layers of asthmatic and nonasthmatic control subjects. The number of cells within the epithelium and subepithelium expressing IL-11 mRNA was greater in those with moderate and severe asthma compared with mild asthma and nonasthmatic subjects (P <.001). There were also greater numbers of IL-11 mRNA positive cells within the subepithelium in severe compared with moderate asthma (P <.001). Immunostaining for IL-11 within the airway tissues confirmed translation of the mRNA into IL-11-immunoreactive protein in airway epithelial cells. Colocalization of IL-11 mRNA and immunoreactivity with resident inflammatory cells demonstrated that this cytokine was also expressed by major basic protein-positive eosinophils. CONCLUSION: These results suggest that IL-11 is involved in the chronic remodeling seen in asthmatic airways and is associated with increasing severity of the disease. PMID- 10669842 TI - Bronchoalveolar lavage in bronchiolitis obliterans organizing pneumonia primed by radiation therapy to the breast. AB - BACKGROUND: Growing evidence indicates that unilateral lung irradiation for breast cancer may "prime" the development of migratory lung infiltrates with histologic features of bronchiolitis obliterans organizing pneumonia. OBJECTIVE: Our purpose was to evaluate the cytologic and immunocytologic features of bronchoalveolar lavage in this condition. METHODS: We analyzed the profile bronchoalveolar lavage cell differentials and lymphocyte subpopulations of 11 women with bronchiolitis obliterans organizing pneumonia syndrome after radiation therapy for breast cancer in comparison to 9 healthy women. RESULTS: The bronchoalveolar lavage analysis demonstrated a significant increase in the percentage of lymphocytes (36.7% +/- 5.4% vs 8.6% +/- 1.1%, P =.0002), neutrophils (3.8% +/- 1.2% vs 0.6% +/- 0.2%, P =.005), eosinophils (2.4% +/- 1% vs 0.3% +/- 0.1%, P =.01), and mast cells (1.4% +/- 0.6% vs 0.1% +/- 0.02%, P =.05) with a significant decrease in the percentage of macrophages (56.1% +/- 6% vs 90.3% +/- 1.4%, P =.0002) in patients with bronchiolitis obliterans organizing pneumonia compared with the control subjects. The percentage of CD3(+) cells was significantly increased in patients with bronchiolitis obliterans organizing pneumonia (93.7% +/- 1.3% vs 70.9% +/- 4%, P =.0004), with a significant decrease in CD4(+) cells (32.7% +/- 4.7% vs 55.4% +/- 2. 6%, P =.002) and a significant increase in CD8(+) cells (61.2% +/- 4. 8% vs 37.5% +/- 2.9%, P =.003) in comparison to control subjects. The CD4/CD8 ratio was significantly reduced in patients with bronchiolitis obliterans organizing pneumonia compared with control subjects (0.6% +/- 0.1% vs 1.5% +/- 0.1%, P =.001). CONCLUSION: These data add to the view that unilateral lung irradiation for breast cancer may "prime" the development of a syndrome quite similar to idiopathic bronchiolitis obliterans organizing pneumonia. PMID- 10669843 TI - Evaluation of a short form for measuring health-related quality of life among pediatric asthma patients. AB - BACKGROUND: This study was undertaken to derive and validate a short form parent completed questionnaire to measure health-related quality of life (HRQL) in pediatric asthma patients. OBJECTIVE: The objectives of this study were to (1) use stepwise analysis to derive a shorter questionnaire from the original long form questionnaire and (2) determine the tradeoff in precision between the long- and short-form surveys. METHODS: One hundred eighty-one pediatric asthma patients were enrolled from 4 sites. A parent of each patient completed a general and an asthma-specific questionnaire during routine office visits from June 1995 to January 1997. The questionnaire included the Child Health Questionnaire Parent Form 50, a general HRQL survey, and a 17-item asthma-specific battery assessing daytime symptoms, nighttime symptoms, and functional limitations. All scales were scored from 0 to 100, with higher scores indicating better HRQL. Analysis of variance models were used to derive short-form scales from the 17-item long-form scales, and the final asthma-specific short-form scale structure was confirmed with use of stepwise regression. Scale reliability was assessed with Cronbach's alpha. Validity of the short-form questionnaire was assessed by comparing mean scale scores according to the level of asthma severity defined by several clinical criteria. Asthma severity was assessed with use of percent predicted FEV(1), frequency and type of symptoms, parent rating of disease severity, physician rating of disease severity, and resource use (emergency department use and hospitalizations). The relative validity of each of the short-form scales was measured by comparing the proportion of variance explained by each of the short form scales compared with the respective long-form scales. RESULTS: The 17-item asthma-specific battery was reduced to 8 items, the Integrated Therapeutics Group Child Asthma Short Form. The daytime and nighttime symptom scales for each contain 2 items and the functional limitations scale 4 items. Reliability was greater than 0.70 for each of the short-form scales. The absence of ceiling and floor effects indicates each scale's ability to detect changes at both low and high levels of functioning. Lower (poorer) mean HRQL scores for severe cases compared with mild cases, for all disease severity indicators, demonstrated clinical validity. Relative validity estimates, comparing the proportion of explained variance of the short-form scales with that of the long-form scales, ranged from 0. 85 to 1.20, indicating a similar ability to measure change. CONCLUSIONS: This study documents the development of a brief, multidimensional, 8 item questionnaire for measuring HRQL in pediatric asthma patients. The brevity of the questionnaire makes it practical for use in practice settings and to monitor patients. PMID- 10669844 TI - Manchester Asthma and Allergy Study: low-allergen environment can be achieved and maintained during pregnancy and in early life. AB - BACKGROUND: Early exposure to dust mite allergens may be critical for primary sensitization. Reducing exposure may offer a realistic chance for primary prevention of sensitization and asthma, but it is essential to implement measures that can achieve and maintain the low-allergen environment. OBJECTIVE: Our purpose was to assess the effectiveness of mite allergen avoidance measures in achieving and maintaining a low-allergen environment during pregnancy and in the first year of life. METHODS: The Manchester Asthma and Allergy Study is a prospective, prenatally randomized study that follows the development of asthma and atopy in a cohort of infants at high risk (both parents atopic) who are randomly allocated to full mite allergen avoidance or to a normal regimen. Avoidance measures comprise (1) mite-proof covers (mattress, pillow, and quilt) for parental bed, (2) high-filtration vacuum cleaner, (3) vinyl flooring in infant's bedroom, (4) new crib and portable crib mattresses encased in mite-proof material, (5) benzyl benzoate (Acarosan) applied on carpets and soft furniture, (6) bed linens washed in hot water weekly, and (7) washable soft toys. Dust samples from the parental bed, bedroom floor, living room floor, infant's mattress, and nursery floor were collected between the 10th and 14th weeks of pregnancy, immediately after birth, and then at age 6 months and 1 year, and Der p 1 levels were determined by mAb-based ELISA. RESULTS: Recovered Der p 1 from maternal mattress was reduced by 97. 25% (95% confidence interval [CI] 95.25% 98.41%) during the second and third trimesters of pregnancy, with the effect persisting for 6 months (98% reduction, 95% CI 97.25%-99.1%) and 12 months (97.6% reduction, 95% CI 95.7%-98.6%) after the birth (active vs control, P <.000001). Total Der p 1 from bedroom floor in the active group was reduced by 53.7% (95% CI 25.7%-71.2%) in samples collected within 4 weeks of the child's birth, with the percentage reduction being 62. 8% (95% CI 39.3%-77.2%) at 6 months and 26.5% (95% CI -24% to 57.1%) at 1 year (active compared vs control, P <.007). Der p 1 levels in crib mattress and nursery floor in the active group were extremely low (crib mattresses geometric mean [95% CI] 2.3 ng [1.6-3.4] at birth, 6.8 ng [4.5-10] at age 6 months, and 15.6 ng [9.8-24.8] at age 1 year [active vs control, P =.001]; nursery 1 ng [0.9-1.1] at birth, 1.7 ng [1.2-2.5] at age 6 months, and 2 ng [1.3 3.5] at age 1 year [active vs control, P <.00001]). The total amount of allergen recovered at age 1 year was 29-fold (95% CI 15.1- to 56.7-fold) higher in the control group than in the active group. CONCLUSIONS: The avoidance measures used in this study achieved and maintained a low mite allergen environment during pregnancy and in the first year of life in homes of infants at risk of atopy. PMID- 10669845 TI - Longitudinal growth in infants and young children treated with budesonide inhalation suspension for persistent asthma. AB - BACKGROUND: Results of recent growth studies suggest that inhaled glucocorticosteroids may affect growth in children. OBJECTIVE: Three 52-week, open-label extension studies (studies A, B, and C) were conducted to compare the effects of budesonide inhalation suspension (BIS) with conventional asthma therapy (CAT) on long-term safety, including intermediate-term growth, in 3 different pediatric asthma populations. METHODS: Pediatric asthma patients (ages 6 months to 8 years) from 3 multicenter, randomized, 12-week, double-blind, placebo-controlled studies were eligible to enroll in the 52-week, open-label extension studies. The extension studies were multicenter, randomized, open label, active-controlled, parallel-group studies performed at 26 centers in the United States. Subjects in each extension study were randomized in a 2:1 ratio to receive either BIS or CAT. BIS was initially administered at a dose of 0.5 mg once (studies A and C) or twice daily (study B), with attempts made at each clinical visit to gradually reduce the dose to the minimum effective dose that maintains asthma control, as judged by the investigator. CAT consisted of any available therapy for asthma, including inhaled glucocorticosteroids in studies B and C only. Height SD scores, growth velocity, and skeletal age (only in studies B and C) were examined. RESULTS: In total, 670 subjects were randomized; 223 subjects received CAT and 447 received BIS. Mean ages at entry were 63.0 months and 60.9 months in CAT and BIS groups, respectively. Median total daily doses of BIS ranged from 0.5 to 1. 0 mg and the mean duration of treatment exposure was 304 +/- 119 days and 342 +/- 83 days in CAT and BIS groups, respectively. Changes in height SD scores differed significantly between the BIS and CAT groups in study A (-0.19, P =.003), and there was a small, statistically significant decrease in growth velocity (-0.8 cm/y, P =.002) in the BIS-treated group compared with the CAT group. No significant differences were observed between BIS and CAT groups in the changes in height SD scores or in growth velocities in studies B (+0.10 and +0.7 cm/y, respectively) and C (+0.12 and +0.8 cm/y, respectively). No differences in skeletal age were observed between BIS and CAT groups in studies B and C. CONCLUSION: There was a small, statistically significant decrease in growth velocity in the BIS-treated group compared with the CAT group in the study (study A) where inhaled glucocorticosteroid use was prohibited before entry and in the CAT group during the study. In the studies (B and C) where inhaled glucocorticosteroids were allowed in the CAT group, no differences were observed in height SD scores or growth velocity. The clinical relevance of these effects, including impact on final adult height, remain to be determined in prospectively planned studies that assess growth in children. PMID- 10669846 TI - Carbon monoxide is endogenously produced in the human nose and paranasal sinuses. AB - BACKGROUND: Carbon monoxide (CO) has recently emerged as an endogenously produced gaseous mediator that, like nitric oxide (NO), appears to be involved in both upper and lower airway inflammation. In healthy subjects a large part of the exhaled NO seems to originate from the nasal airways, and the paranasal sinuses have been described as a dominating site for NO production. OBJECTIVE: The current study was designed to investigate whether CO could also be produced in the nose and paranasal sinuses. METHODS: The occurrence in the nasal mucosa of the enzyme heme oxygenase, the rate limiting step for CO production, was analyzed with use of immunocytochemistry. CO in exhaled and sampled air was measured with an infrared analyzer. Forty-two healthy subjects and two patients with a tracheostoma volunteered for the study. RESULTS: Heme oxygenase-like immunoreactivity was seen in the respiratory epithelium, in connection with seromucous glands and in the vascular smooth muscle of the nose. When CO was continuously sampled from one nostril during normal breathing through the mouth, stable levels of CO could be measured within 40 seconds in all subjects tested (n = 33). Repeated measurements indicated only minor variations in the values obtained. Sampling through a drainage tube inserted into the maxillary sinus revealed CO levels comparable to the levels obtained by sampling through the nose (n = 6). Breathing through the nose increased the CO levels obtained in the exhaled air (n = 33, P <. 001). CONCLUSION: These results imply that the nose and paranasal sinuses contribute to the CO production of the human airways. PMID- 10669847 TI - Does allergy in parents depend on allergy in their children? Recall bias in parental questioning of atopic diseases. Multicenter Allergy Study Group. AB - BACKGROUND: A positive atopic family history has proved to be one important risk factor for the development of atopic diseases in offspring. However, many epidemiologists are concerned about the accuracy and reliability of data because responses to questionnaires can be biased for many reasons. OBJECTIVE: The study investigated whether responses of parents questioned about their atopic diseases change depending on the development of atopic symptoms in their children. METHODS: During a prospective birth cohort study on atopy in children (the Multicenter Allergy Study) parents filled out questionnaires twice within 2 years about their atopic diseases. Differences between the 2 responses were examined by log-linear and logistic regression models depending on the diagnosed atopy status of the study children. RESULTS: Mothers tended to report more atopic diseases in the second questioning than in the first, indicating a nondifferential misclassification. Fathers were influenced by the development of atopic diseases in their children: they reported significantly more atopic diseases if the child developed atopic illness with atopic dermatitis. CONCLUSION: In parental questioning about atopic diseases, a recall bias must be considered for the association of atopic family history and atopy in children. Especially in case control and cross-sectional studies, such misclassifications can result in biased estimates of prognosis and risk factors. PMID- 10669848 TI - Escherichia coli expression and purification of recombinant dog albumin, a cross reactive animal allergen. AB - BACKGROUND: Animal hair-dander represents an important source of indoor allergens. Diagnosis and therapy of animal allergy would benefit from the availability of defined recombinant allergens. They may be preferred to animal derived proteins, particularly for in vivo application in patients. OBJECTIVE: The purpose of this study was to express and purify recombinant dog albumin, an important cross-reactive animal allergen. METHODS: Complementary (c)DNA sequences coding for dog albumin were obtained by reverse transcription and subsequent PCR amplification from dog liver RNA. Dog albumin-encoding cDNA sequences were inserted into phage lambdagt11, and IgE-reactive phage clones were isolated by immunoscreening with serum IgE from a patient with dog allergy. Dog albumin was expressed as IgE-reactive recombinant protein in Escherichia coli and purified by inclusion body preparation, resolubilization, and diethylaminoethyl cellulose chromatography. Cross-reactivity of dog albumin with cat and human albumin was examined by immunoblot, as well as ELISA inhibition experiments. RESULTS: A cDNA sequence coding for complete dog albumin was obtained by reverse transcription and subsequent PCR amplification from dog liver. The cDNA and deduced amino acid sequence of dog albumin was highly homologous to the sequences of albumins from animals to human subjects, thus explaining the extensive cross-reactivities among albumins. Recombinant dog albumin was expressed in E coli and purified. It reacted with serum IgE from patients allergic to dog albumin and a monoclonal anti-human albumin antibody. Immunologic competition experiments performed with serum IgE from patients allergic to dog albumin and a mouse monoclonal antihuman albumin antibody indicated the presence of similar epitopes on dog, cat, and human albumin. CONCLUSION: Recombinant dog albumin may be used for diagnostic purposes to identify patients who are cross-sensitized to many animal species and perhaps for specific immunotherapy of sensitized individuals. PMID- 10669849 TI - Purification and characterization of an 18-kd allergen of birch (Betula verrucosa) pollen: identification as a cyclophilin. AB - BACKGROUND: Five birch pollen allergens have been identified so far. In a previous study we detected new birch pollen allergens with an isoelectric point in the range 9.0 to 9.3, present only in extracts prepared at controlled basic pH. OBJECTIVE: The purpose of the current study was to purify and characterize those allergens. METHODS: The target allergens were purified by ion exchange and hydrophobic interaction chromatography. Analyses were carried out by SDS-PAGE, isoelectric focusing, immunoblotting, and amino acid sequencing. The in vivo reactivity of the allergens was evaluated by skin testing. RESULTS: An 18-kd protein, which we named Bet v 7, was purified. This 18-kd protein corresponded to 3 bands on isoelectric-focusing immunoblots that probably represent isoforms. On immunoblots up to 20.8% of birch pollen-allergic patients recognized those allergens. The clinical relevance of Bet v 7 was demonstrated by positive immediate-type skin testing on a patient allergic to birch pollen. Sequencing of an internal peptide yielded an amino acid sequence showing high homology with various plant cyclophilins. The rotamase activity of the protein, inhibited by cyclosporin A, further confirmed that Bet v 7 belongs to the group of cyclophilins. CONCLUSION: We have purified a novel allergen of birch pollen, Bet v 7, belonging to the cyclophilin family. Because cyclophilins are highly conserved proteins over the phylogeny, we may postulate that Bet v 7 is a member of a new family of panallergens. PMID- 10669850 TI - Genetic restrictions in olive pollen allergy. AB - BACKGROUND: The major antigen of olive tree pollen, Ole e 1, produces an IgE response restricted by DQ2. OBJECTIVE: Our purpose was to further analyze the genetic restrictions associated with IgE and IgG antibodies against Ole e 1 and IgE against the recently described antigen Ole e 3. METHODS: Twenty-two nuclear olive pollen-allergic families (n = 88) were selected. DRB1 and DQB1, TCR-Valpha 8.1, the high-affinity receptor of IgE (FcepsilonRI-beta) Rsa I exon 7 and intron 2 and TNF-beta (LTalpha-Nco I) polymorphisms were determined by PCR and analyzed for association with allergic traits by the multiallelic transmission disequilibrium test. RESULTS: Significant associations were found among HLA DQB1*0201 (n = 29) and high levels of IgG (P =.023) and IgE (P =.0136) antibodies to Ole e 1 and with IgE specific to Ole e 3 (P =.0368). DRB1*0701 was associated with high levels of total serum IgE (P =.04) and IgG against Ole e 1 (P =.025). The FcepsilonRI-beta Rsa I exon 7, allele 1 (n = 39), was associated with high levels of total serum IgE (P =. 01), IgE antibodies against Olea europaea extract (P =.004), and specific antibodies to Ole e 1, IgG (P =.04), and IgE (P =.006). The FcepsilonRI-beta Rsa I intron 2, allele 2 (n = 33), was associated with IgE antibodies to O europaea extract (P =.003) and specific antibodies to Ole e 1, IgG (P =.025), and IgE (P =.05). CONCLUSIONS: We found a new association between IgE antibody response to Ole e 3 and DQB1*0201 and verified the previously reported association between Ole e 1-specific response and DQB1*0201. Also, the association between FcepsilonRI-beta and IgE antibodies against Ole e 1 was demonstrated. PMID- 10669851 TI - The role of IL-12 in the induction of late-phase cellular infiltration in a murine model of allergic conjunctivitis. AB - BACKGROUND: The applied murine model of allergic conjunctivitis mimics human disease, and an immediate hypersensitivity reaction (IHR) and a late-phase cellular reaction typically develop in sensitized mice after topical challenge with the allergen. OBJECTIVE: We investigated the role of IL-4, IFN-gamma, and IL 12 in the early and late phases of ocular allergy with use of cytokine knockout (KO) mice and neutralizing antibodies. METHODS: Ragweed-sensitized wild-type or IL-4KO, IL-12KO, IFN-gamma KO, anti-IL-12 mAb-treated, recombinant murine IL-12 treated, and anti-IFN-gamma mAb-treated mice were challenged with the allergen 10 days after the immunization. IHR, cellular infiltration, lymphoproliferative response, and cytokine production from draining lymph nodes were recorded and compared among groups. RESULTS: We show that IL-12KO mice and anti-IL-12 antibody treated wild-type animals failed to have a cellular infiltration into the conjunctiva. Treatment with recombinant murine IL-12 also reduced the number of infiltrating PMNs but increased the percentage of mononuclear cells in the conjunctiva compared with controls. IFN-gamma KO mice had a significantly stronger IHR and prolonged infiltration into the conjunctiva after challenge with ragweed than controls. CONCLUSION: Our data suggest that the presence of IL-12, although better known as a T(H)1-inducing cytokine, is important for the development and the regulation of the late-phase pathologic features in ocular allergy. Furthermore, IFN-gamma is a limiting factor in the late phase of allergy and thus may be important in preventing chronic allergic disease. PMID- 10669852 TI - Blood eosinophils from atopic donors express messenger RNA for the alpha, beta, and gamma subunits of the high-affinity IgE receptor (Fc epsilon RI) and intracellular, but not cell surface, alpha subunit protein. AB - BACKGROUND: Blood eosinophils from hypereosinophilic donors were previously reported to possess the functional high-affinity IgE receptor (Fc epsilon RI), so providing a potential mechanism to account for eosinophil degranulation in atopic allergic disease. Furthermore, tissue eosinophils from allergic tissue reactions were shown to be mRNA(+) for the alpha, beta, and gamma subunits of Fc epsilon RI and gave positive immunostaining with an anti-Fc epsilon RI-alpha antibody. Recent studies, however, revealed negative surface staining on peripheral blood eosinophils, but intracellular Fc epsilon RI-alpha protein was identified by Western blot analysis. OBJECTIVE: Our purpose was to examine on peripheral blood eosinophils from atopic subjects (1) surface expression and mRNA for Fc epsilon RI-alpha, (2) up-regulation of Fc epsilon RI-alpha by allergy-associated tissue factors, and (3) Fc epsilon RI-alpha-dependent release of eosinophil peroxidase (EPO). METHODS: We measured (1) Fc epsilon RI mRNA expression by in situ hybridization, (2) Fc epsilon RI-alpha by flow cytometry and immunocytochemistry (with use of nonpermeabilized and permeabilized cells), and (3) Fc epsilon RI alpha-dependent release of EPO. RESULTS: Eosinophils from atopic donors had negligible surface expression of Fc epsilon RI-alpha, which was not enhanced by culture with IgE, IL-3, IL-4, IL-5, GM-CSF, or fibronectin or coculture with fibroblasts. Permeabilization, however, revealed appreciable intracellular staining for Fc epsilon RI-alpha. The majority of eosinophils were mRNA(+) for the alpha, beta, and gamma subunits of Fc epsilon RI. Small but significant (P =.03) increases in alpha chain mRNA expression were observed after coculture of eosinophils with fibroblasts but not with IgE, IL-4, or fibronectin. Cross linking of Fc epsilon RI on the surface of eosinophils from atopic donors did not lead to detectable EPO release. CONCLUSION: Human blood eosinophils express negligible, nonfunctional membrane Fc epsilon RI-alpha but have intracellular Fc epsilon RI-alpha protein and mRNA expression for the alpha, beta, and gamma subunits. PMID- 10669853 TI - Corticosteroids inhibit rhinovirus-induced intercellular adhesion molecule-1 up regulation and promoter activation on respiratory epithelial cells. AB - BACKGROUND: Rhinoviruses are associated with the majority of asthma exacerbations. To date, the pathogenesis of virus-induced asthma exacerbations is still unclear, and no safe effective therapy is available. Intercellular adhesion molecule-1 (ICAM-1) has a central role in inflammatory cell recruitment to the airways in asthma and is the receptor for 90% of rhinoviruses. We have previously shown that rhinovirus infection of lower airway epithelium induces ICAM-1 expression by a transcriptional mechanism that is critically nuclear factor kappaB-dependent. OBJECTIVE: The purpose of this study was to investigate the effect of systemic (hydrocortisone [HC], dexamethasone [DM]) and topical (mometasone furoate [MF]) corticosteroids on rhinovirus-induced ICAM-1 up regulation. METHODS: Cultured primary bronchial or transformed (A549) respiratory epithelial cells were pretreated with corticosteroids for 16 hours and infected with rhinovirus type 16 for 8 hours. ICAM-1 surface expression was evaluated by flow cytometry. In A549 cells ICAM-1 messenger RNA was evaluated by specific reverse transcription-PCR and promoter activation by chloramphenicol acetyltransferase assay. RESULTS: We observed inhibition of rhinovirus-induced ICAM-1 up-regulation with corticosteroid pretreatment in both primary bronchial epithelial and A549 cells. In A549 cells systemic and topical corticosteroids demonstrated a dose-dependent inhibition with similar efficacy (inhibitory concentration 50% 10(-10) mol/L, 10(-11) mol/L, and 10(-11) mol/L for HC, DM, and MF respectively). MF also inhibited ICAM-1 messenger RNA induction by rhinovirus infection in a dose-dependent manner. MF completely inhibited rhinovirus-induced ICAM-1 promoter activation. HC, DM, and MF had no direct effect on rhinovirus infectivity and replication in cultured cells. CONCLUSION: Corticosteroids decrease rhinovirus-induced ICAM-1 up-regulation in respiratory epithelial cells and modulate pretranscriptional mechanisms. This effect may be important for the therapeutic control of virus-induced asthma exacerbations. PMID- 10669854 TI - IL-4 production by PBMCs on stimulation with mite allergen is correlated with the level of serum IgE antibody against mite in children with bronchial asthma. AB - BACKGROUND: Although IL-4, IL-13, and IFN-gamma are known to affect IgE synthesis, it remains unclear which one plays the most important role in in vivo IgE synthesis in atopic patients. OBJECTIVE: The aim of this study is to clarify the difference in importance among these cytokines in up-regulation of IgE synthesis in atopic patients. METHODS: We measured IL-4, IL-13, and IFN-gamma production by PBMCs on stimulation with house dust mite (HDM) in 23 children, 3 to 15 years old, with bronchial asthma (BA) and analyzed the correlation with HDM specific IgE antibody levels expressed as HDM IgE radioallergosorbent test (RAST) results. RESULTS: The production of IL-4 and IL-13 by PBMCs on stimulation with HDM was significantly higher in children with BA than in nonatopic control subjects (IL-4, 752.9 +/- 365.9 vs 312.3 +/- 230.0 fg/mL, P <.001; IL-13, 21.9 pg/ml [<12.0-77.6] vs <12.0, P <.01). IL-4 production showed a close positive correlation with HDM IgE RAST (r = 0.71, P <.001), which was distinctly stronger than that between IL-13 production and HDM IgE RAST (r = 0.46, P <.05). IFN-gamma production was neither different between children with BA and nonatopic control subjects (7. 24 [1.54-33.90] pg/mL vs 11.2 [1.66-75.9] pg/mL) nor correlated with HDM IgE RAST levels. Essentially the same result was obtained by stimulation of PBMCs with a purified HDM major allergen Der f 1. CONCLUSION: IL-4 is likely to be the most important cytokine in up-regulation of in vivo IgE synthesis against HDM in children with BA. PMID- 10669856 TI - In vitro diagnosis of cypress pollen allergy by using cytofluorimetric analysis of basophils (Basotest). AB - BACKGROUND: Cupressaceae pollen allergy is a worldwide pollinosis, but its in vitro diagnosis is notoriously difficult. The Basotest is a newly available in vitro test for the detection of allergen-specific IgE based on the level of cellular activation of basophils by using flow cytometry. OBJECTIVES: The Basotest was compared with the measurement of cypress pollen-specific IgE in highly selected patients. METHODS: We analyzed 34 patients allergic to cypress pollen selected on the basis of a suggestive clinical history and positive skin test and nasal challenge responses to cypress pollen extract. We also analyzed 8 patients with positive skin test responses to cypress pollen extract who did not present symptoms during the pollen season (intermediate group) and 33 control subjects. Sensitivity, specificity, and efficiency of the Basotest and serum specific IgE levels measured by using the CAP System were determined in patients allergic to cypress pollen. Histamine release was studied in a selected group of patients. RESULTS: The Basotest was more sensitive (91.2%) than the CAP System (76%) for the in vitro diagnosis of cypress pollen allergy. A dose-response curve was observed in basophils obtained from patients allergic to cypress pollen. There were no false-positive results with either test (specificity 100%). The results of the Basotest or those of the CAP System did not correlate with the patients' in vivo threshold sensitivity assessed by skin tests and nasal challenge. CONCLUSIONS: The Basotest was found to be an effective diagnostic test in patients allergic to cypress pollen. PMID- 10669855 TI - Chemokine- and cytokine-induced expression of endothelin 1 and endothelin converting enzyme 1 in endothelial cells. AB - BACKGROUND: Endothelin 1 (ET-1) is a product of endothelial and many other cell types that possesses a wide range of actions, including vasoconstriction, bronchoconstriction, and mitogenic activity on smooth muscle cells and fibroblasts. ET-1 release and expression is induced in several disease conditions associated with inflammation and cellular injury. OBJECTIVE: The purpose of this study is to determine the effects of alpha-chemokines (IL-8 and melanoma growth stimulating activator), beta-chemokines (monocyte chemotactic protein 1, macrophage inflammatory protein 1alpha [MIP-1alpha], MIP-1beta, and RANTES), and proinflammatory cytokines (IL-1beta, TNF-alpha, and IFN-gamma) on the expression of both ET-1 and endothelin-converting enzyme 1 (ECE-1) by human umbilical vein endothelial cells. METHODS: Subconfluent monolayers of human umbilical vein endothelial cells were incubated with each chemokine individually for 24 hours or with a mixture (cytomix) of TNF-alpha, IL-1beta, and IFN-gamma for 6 and 24 hours. RESULTS: Incubation with the alpha-chemokines melanoma growth-stimulating activity and IL-8 did not significantly increase ET-1 and ECE-1 messenger (m)RNA expression and had no effect on ET-1 release. Monocyte chemotactic protein 1 exerted the most potent increase in ET-1 and ECE-1 mRNA and ET-1 release among all chemokines studied (P <.05). MIP-1alpha and RANTES exerted a moderate, but significant, increase on the ET system (P <.05). The cytomix resulted in a significant increase in ET-1 and ECE-1 mRNA expression (P <.05). CONCLUSION: These data demonstrate that, like cytokines, chemokines can induce endothelial ET 1 and ECE-1 in vitro and suggest a possible role for these inflammatory mediators in the induction of the ET system in inflammatory and vascular diseases. PMID- 10669857 TI - Helper T-cell responses elicited by Der p 1-pulsed dendritic cells and recombinant IL-12 in atopic and healthy subjects. AB - BACKGROUND: Environmental allergens, such as Dermatophagoides pteronyssinus group 1 antigen (Der p 1), induce T(H2)-type responses in atopic patients, whereas healthy individuals have T(H1)-type responses to the same antigens. Because of their efficient synthesis of IL-12, dendritic cells (DCs) are potent inducers of T(H1)-type immune responses. OBJECTIVE: We sought to determine whether DCs would skew allergen-specific T(H2)-type responses from atopic individuals. METHODS: Purified CD4(+) T cells from healthy donors or atopic individuals were cultured in the absence or presence of recombinant (r)IL-12 with DCs derived from PBMCs and pulsed with Der p 1. Supernatants of DC-T cell cocultures were assayed by ELISA for IL-5 and IFN-gamma. RESULTS: A T(H1)-type response developed in purified CD4(+) T cells from healthy donors in response to Der p 1-pulsed DCs, as indicated by high levels of IFN-gamma in culture supernatants. In contrast, CD4(+) T cells from atopic donors displayed a T(H2)-type profile characterized by high levels of IL-5 and low levels of IFN-gamma. The addition of rIL-12 (10 ng/mL) to DC-T cell cocultures resulted in the induction of IFN-gamma secretion by Der p 1-specific CD4(+) T cells from atopic patients, whereas their production of IL-5 was not inhibited. Using flow cytometry after intracytoplasmic staining, we found that IFN-gamma and IL-5 were secreted by distinct CD4(+) T-cell subpopulations. CONCLUSION: The cytokine profile of Der p 1-specific T(H2)-like cells from atopic individuals is maintained when the allergen is presented by DCs, even in the presence of exogenous rIL-12. PMID- 10669858 TI - Urinary eosinophil protein X and serum eosinophil cationic protein in infants and young children with atopic dermatitis: correlation with disease activity. AB - BACKGROUND: Eosinophil cationic protein (ECP) and eosinophil protein X (EPX) or eosinophil-derived neurotoxin (EDN) are released by eosinophil granulocytes in allergic diseases. Serum ECP (s-ECP) levels have been correlated with disease activity in atopic dermatitis (AD) in adults and young patients, and high urinary EPX (u-EPX) levels in asthmatic patients seem to reflect active disease. A relationship between AD severity and u-EPX concentration in young children has not been previously studied. OBJECTIVE: This study was performed to evaluate whether the severity of AD in infants and young children was correlated with s ECP and u-EPX levels. METHODS: Fifty-four infants and children (mean age, 17.7 months; range, 4-48 months) with AD and without other allergic conditions were evaluated. The severity of AD was measured by using the SCORAD index. S-ECP, serum total IgE, serum-specific IgE for common allergens, and peripheral blood eosinophil counts (PBECs) were determined. In forty-two children u-EPX was also measured. Seven age-matched control patients underwent the same determinations. RESULTS: S-ECP and u-EPX were significantly higher in children with AD than in control children (mean, 23.9 vs 3.5 microg/dL [P <.001] and 57.7 vs 6.0 microg/mmol creatinine [P <.001]). A significant correlation was found between SCORAD and s-ECP (P =.002), u-EPX (P =.01), and PBECs (P =.01) and between symptom index and uEPX (P =.0004). PBECs were strongly correlated to s-ECP and u EPX (P <.0001). However, 5 patients with moderate and severe AD (11.9%) showed low levels of s-ECP, u-EPX, and PBECs. CONCLUSION: S-ECP and u-EPX were useful markers of AD activity in infants and young children. When taken together, the two determinations could give more information about the clinical course of the illness. Some patients seemed to have clinical exacerbations without an involvement of eosinophils and their products. PMID- 10669859 TI - Carbon dioxide inhalation challenges in idiopathic environmental intolerance. AB - BACKGROUND: Idiopathic environmental intolerance (IEI) is associated with unexplained physical symptoms, which overlap considerably with those of panic disorder (PD). OBJECTIVE: This study tested the hypothesis that patients with symptoms to suggest IEI exhibit features of PD in response to nonnoxious environmental stimuli. METHODS: A single-blind, case-control 35% carbon dioxide inhalation challenge was conducted at a university-based occupational health unit with the use of standardized psychologic questionnaires involving 36 patients with IEI and 37 healthy control subjects. The main outcome measures included panic attack symptoms and scores on the Anxiety Sensitivity Index, a measure of panic-related anxiety. RESULTS: Patients with IEI scored significantly higher on the Anxiety Sensitivity Index than control subjects did (P <.05). Significantly more patients with IEI (71%) than control subjects (26%) fulfilled panic attack criteria after carbon dioxide (P <.001). Physiologic responses to the challenge were not significantly different between groups. CONCLUSIONS: Results suggest that, similar to patients with PD, patients with IEI display high anxiety sensitivity and in response to carbon dioxide inhalation tend to experience heightened anxiety and panic attacks. PMID- 10669860 TI - Exposure-effect relationship of platinum salt allergy in a catalyst production plant: conclusions from a 5-year prospective cohort study. AB - BACKGROUND: There is a high incidence of occupational asthma and rhinitis caused by platinum (Pt) salts in precious-metal refineries. OBJECTIVE: We sought to assess exposure to Pt salts and the incidence of Pt salt allergy in a catalyst production plant. METHODS: A 5-year prospective cohort study was performed in 159 catalyst production workers (94.6% of recruited), 50 craftsmen (92. 6% of recruited), and 66 control subjects (76.7% of recruited) at yearly intervals. Subjects were assigned to exposure categories of high levels of Pt (n = 115), persistently low levels of Pt (n = 51), intermittently low levels of Pt (n = 61), or no Pt (n = 48) after the initial survey according to job title and job location. Skin prick test conversion from a negative response to a 4 mm or larger wheal response with a 10(-2) mol/L hexachloroplatinic acid solution was chosen as the outcome variable. RESULTS: Exposure assessment of airborne Pt and Pt in the serum of workers demonstrated clear differences between exposure categories. The threshold limit value of 2 microg/m(3) for soluble Pt was exceeded in 3 (4%) of 78 measurements. Thirteen subjects assigned to high exposure showed skin test conversion, and new allergic symptoms were associated with exposure. Among the high-exposure category, the incidence rate of skin prick test conversion was 5.9 per 100 person-years for newly employed subjects (n = 79) and 2.1 per 100 person years for those who had already been employed at the time of the initial survey (n = 36). A predicting factor for skin test conversion in highly exposed subjects was smoking status (relative risk, 3.9; 95% confidence interval, 1.6-9.7) but not atopy or bronchial hyperresponsiveness. CONCLUSION: Sensitization to Pt salts may develop in workers of catalyst production plants. Both the exposure to Pt salts and the incidence of Pt salt allergy were lower compared with reported data from precious-metal refineries. PMID- 10669861 TI - Evidence for a lipid transfer protein as the major allergen of apricot. AB - BACKGROUND: Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach. PMID- 10669862 TI - Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K. AB - BACKGROUND: Peanuts and soybeans are 2 foods that have been shown to be responsible for many atopic disorders. Because of their nutritional benefit, soybean proteins are now being used increasingly in a number of food products. Previous studies have documented multiple allergens in soybean extracts, including glycinin, beta-conglycinin, and the P34/Gly m Bd 30K protein. OBJECTIVE: Our overall goal was to identify soybean-specific allergens to begin to understand molecular and immunochemical characteristics of legume proteins. The specific aim of the current investigation was to identify the essential amino acid residues necessary for IgE binding in the 5 distinct immunodominant epitopes of P34/Gly m Bd 30K. METHODS: Serum IgE from 6 clinically sensitive soybean allergic individuals was used to identify P34/Gly m Bd 30K in the native and single amino acid substituted peptides with use of the SPOTS peptide synthesis technique to determine critical amino acids required for IgE binding. RESULTS: The intensity of IgE binding and epitope recognition by serum IgE from the individuals varied substantially. With use of serum from 6 clinically soybean sensitive individuals, 2 of the 5 immunodominant epitopes could be mutagenized to non-IgE binding peptides. CONCLUSIONS: Single-site amino acid substitution of the 5 immunodominant epitopes of Gly m Bd 30K with alanine revealed that IgE binding could be reduced or eliminated in epitopes 6 and 16 in the serum obtained from 6 soybean-sensitive patients. PMID- 10669863 TI - Survey of patients after discontinuing venom immunotherapy. AB - BACKGROUND: Venom immunotherapy rapidly reduces the risk of a systemic sting reaction in adults from 30% to 70% to less than 2%. When venom immunotherapy is stopped after 5 years or longer, the risk of a systemic sting reaction is 5% to 15% during the first few years after stopping treatment. It is uncertain whether systemic sting reactions will occur more than 5 years after discontinuing venom immunotherapy and whether treatment can be safely stopped in some patients after less than 5 years. OBJECTIVE: The purpose of this study is to estimate the risk of systemic reaction to a sting 10 years after discontinuing treatment and the relative risk after 3 years of treatment compared with that after 5 years or more of treatment. METHODS: Among all patients who had venom immunotherapy at our center, we identified 395 patients who stopped treatment: some had dropped out of therapy early (6-24 months), some stopped after 3 to 4 years, and most completed 5 years or more of venom immunotherapy and were advised to stop by the allergist (many as part of our reported studies of discontinuing venom immunotherapy). RESULTS: Contact was made with 194 patients, including telephone interviews for sting history and requests to visit the office for skin testing and blood sampling. Of these patients, 74 had been included in our original study of patients who had 5 years or more of venom immunotherapy and had sting challenges after 1 to 5 years off venom immunotherapy, as previously reported. Of the 74 in that original study, 61 were reached for this survey, and 30 reported recent stings, with 5 systemic sting reactions. Another 133 patients who had stopped venom immunotherapy were reached: 82 had 5 or more years of venom immunotherapy, 20 had 3 to 4 years of venom immunotherapy, and 31 had less than 2 years of venom immunotherapy. Of 51 patients stung from this group, 27 had 5 or more years of venom immunotherapy (no systemic sting reactions), and 24 had less than 5 years of venom immunotherapy (3 systemic sting reactions). We have now observed a total of 113 patients who had 5 or more years of venom immunotherapy and were stung after stopping. Sixteen (14%) had systemic sting reactions; most were mild, but 4 were severe. Systemic sting reactions occurred in 12 (10.7%) of 112 patients stung in the first 4 years off venom immunotherapy and 5 (10%) of 50 stung more than 5 years off venom immunotherapy. In 4 of 8 patients with current systemic sting reactions, the skin test response was negative, although the venom-IgE response was positive at the previous encounter. All systemic sting reactions were similar in pattern and severity to prevenom immunotherapy reactions in the same patient. CONCLUSIONS: We conclude that the risk of systemic sting reactions when venom immunotherapy is stopped after 5 years or longer remains in the reported range of 5% to 15% in the 5 to 10 years after stopping venom immunotherapy. This risk of systemic sting reactions does not seem to decrease over time, unlike the progressive decline in immunologic markers (skin test and venom-IgE responses). To prospectively assess the risk of recurrent systemic sting reactions, there is a need for sting challenge studies of patients who have been off venom immunotherapy for 5 to 10 years and patients who have stopped venom immunotherapy after just 3 to 4 years treatment. PMID- 10669864 TI - Lack of correlation between Chlamydia pneumoniae antibody titers and adult-onset asthma. PMID- 10669865 TI - Body build and atopy. PMID- 10669867 TI - Sequence analysis of two cosmids from the right arm of the Schizosaccharomyces pombe chromosome II. AB - We report the complete sequence of two cosmids, SPBC19C7 (34815 bp insert, Accession No. AL023859) and SPBC15D4 (33203 bp insert, Accession No. AL031349), localized on chromosome II of the S. pombe genome. Twelve open reading frames (ORFs) were identified in SPBC19C7 and 16 in SPBC5D4. Two known genes were found on each cosmid: cyr1 and uve1 on SPBC19C7, encoding adenylate cyclase and a UV endonuclease, respectively, and gpt and pho2 on SPBC15D4, encoding an N acetylglucosamine-1-phosphate transferase and a4-nitrophenylphosphatase, respectively. Five ORFs similar to known proteins were found on SPBC19C7, and six on SPBC15D4. They include putative genes for a ubiquitin protein ligase, a prolyl tRNA synthetase, a tRNA splicing endonuclease, a voltage-gated chloride channel, a mannosyl transferase, a kinesin-like protein, a histone transcriptional regulator, an N-acetyltransferase, a cystathionine gamma-synthase and a TFIID subunit. Two ORF products of SPBC15D4 do not have clear homologues: one encodes a putative transcriptional regulator with a binuclear zinc domain and the other a protein with six transmembrane domains. Two ORFs from SPBC15D4 are similar to unknown ORFs, one from Saccharomyces cerevisiae and the other from Caenorhabditis elegans. Finally, two ORFs of SPBC19C7 and six of SPBC15D4 correspond to orphan genes. The frequent occurrence of introns and the short and degenerated intron exon boundaries consensus sequences significantly complicated ORF predictions. Two potential ORF-free regions spanning several kb were predicted, and a clustering of ORFs transcribed in the same orientation was observed. PMID- 10669868 TI - Isolation and molecular characterization of KlCOX14, a gene of Kluyveromyces lactis encoding a protein necessary for the assembly of the cytochrome oxidase complex. AB - The yeast Kluyveromyces lactis was mutagenized with ethyl methane sulphonate and mutants unable to grow on respiratory carbon sources were isolated. Functional complementation of one of these mutants led to the isolation of KlCOX14, a gene encoding a 64 amino acid protein which is the functional homologue of Saccharomyces cerevisiae Cox14p, a protein necessary for the assembly of the cytochrome oxidase holoenzyme (Glerum et al., 1995). The disruption of KlCOX14 resulted in the absence of the absorption bands relative to cytochromes a and a(3) and in the complete loss of respiratory activity. Klcox14 mutants display the typical phenotype of pet mutants and have a reduced growth rate. In addition, unlike the wild-type, Klcox14 mutants are able to grow by fermentation also in the presence of low glucose. The nucleotide sequence of KlCOX14 has been deposited in the EMBL databank with Accession No. AJ238801. PMID- 10669870 TI - CIp10, an efficient and convenient integrating vector for Candida albicans. PMID- 10669869 TI - Secretory production of recombinant human chymase as an active form in Pichia pastoris. AB - We succeeded in expressing in a Pichia pastoris (P. pastoris) host a cDNA encoding a mature human chymase (h-chymase) which was secreted directly into the culture medium. Recombinant human heart chymase (rh-chymase) was purified from the culture medium via a single one-step heparin-agarose column chromatography tracing, using succinyl-Ala-Ala-Pro-Phe-para-nitroanilide (Suc-AAPF-pNA) hydrolysing activity. On SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the rh-chymase showed a diffused protein band with molecular weight of 32-37 kDa. After deglycosylation, however, rh-chymase changed to a sharp protein band with molecular weight 28 kDa, which is equal in size to deglycosylated h-chymase. The rh-chymase had an activity to convert one of the natural substrates, angiotensin I, to angiotensin II. Double reciprocal plot analysis revealed that the K(m) value ofrh-chymase against Suc-AAPF-pNA was approximately 5.1 mM, which is close to that of purified h-chymase. PMID- 10669871 TI - Protein disulphide isomerase genes of Kluyveromyces lactis. AB - Two genes of Kluyveromyces lactis, KlPDI1 and KlMPD1, were studied. They code for a protein disulphide isomerase and its structural and functional homologue, respectively. The KlPDI1 product was 52.6% identical to Pdi1p and the KlMPD1 product 47% identical to Mpd1p of S. cerevisiae. Both genes contained the thioredoxin-active site-related signature. Their C-termini showed a new variant of the endoplasmic reticulum-retention signal, QDEL. A single copy of KlPDI1 was able to complement the growth defect of a pdi1 mutation. KlMPD1 on a multicopy vector partially suppressed the klpdi1 and pdi1 mutations. The Klpdi1 null mutation was lethal. The klmpd1 disruptant was viable, but showed an increased sensitivity to high temperature. Several stress response motifs were present in the upstream sequence of KlMPD1, but not of KlPDI1, whilst the opposite is known for the S. cerevisiae homologues. The viability of the klmpd1 mutant under starvation for nitrogen or carbon source was not different from that of the wild type. The syntenic relationship is discussed for the KlPDI1 gene regions with respect to the duplicated segments PDI1/EUG1 in S. cerevisiae. PMID- 10669872 TI - The ubiquitin-encoding genes of Kluyveromyces lactis. AB - The ubiquitin encoding genes of Kluyveromyces lactis were cloned. Three genes, KlUBI1, KlUBI3 and KlUBI4, were found in this yeast, while in Saccharomyces cerevisiae there are four genes, UBI1, -2, -3 and -4. The UBI1/UBI2 duplication is thus absent from the K. lactis genome. General structural features of ubiquitin genes were very similar in these two species (presence of an intron in KlUBI1, fusion to ribosomal protein genes in KlUBI1 and KlUBI3, spacer-less polyubiquitin repeats in KlUBI4). Disruption or deletion of K. lactis ubiquitin genes showed that: (a) disruption of KlUBI1 was lethal (in S. cerevisiae, ubi1/ubi2 double deletion is lethal); (b) KlUBI3 is also an essential gene for cell growth; (c) deletion of KlUBI4 led to an increased sensitivity to high temperature, similar to the ubi4 mutation in S. cerevisiae, but, in contrast to the latter, the klubi4 mutant was not sensitive to carbon or nitrogen source starvation. The syntenic relationship of ubiquitin loci between K. lactis and S. cerevisiae genomes is also described. PMID- 10669873 TI - Identification of functionally important regions of the Saccharomyces cerevisiae mitochondrial translational activator Cbs1p. AB - Translation of cytochrome b mRNA in yeast mitochondria requires activation by the nuclear-encoded Cbs1p. According to the current model, Cbs1p tethers cytochrome b mRNA to the inner mitochondrial membrane via interaction with the 5'-untranslated leader. Cbs1p is predicted to be a hydrophilic protein with two hydrophobic segments near the carboxyl-terminal end, which are both too short to span the membrane. Nevertheless Cbs1p is tightly associated with the mitochondrial membrane, as shown by its behaviour in extraction experiments with taurodeoxycholate. In an attempt to define functionally important regions of Cbs1p, we created a number of mutant alleles by random and directed mutagenesis. We report that a Cbs1p mutant protein lacking the mitochondrial presequence is still able to complement a Deltacbs1 strain, suggesting that the presequence does not contain essential mitochondrial targeting information. Mutations in a cluster of positively charged amino acids at the extremeC-terminus have no effect on Cbs1p function, but removal of this segment severely impairs Cbs1p function. Truncation of 12 or more amino acids from the C-terminus results in a completely defective protein. We further show that both short hydrophobic regions are essential for Cbs1p function, although membrane association is observed even in the absence of these regions. PMID- 10669874 TI - Disruption and phenotypic analysis of six open reading frames from the left arm of Saccharomyces cerevisiae chromosome VII. AB - Six open reading frames (ORFs) from Saccharomyces cerevisiae chromosome VII were deleted using the kanMX4 module and the long-flanking homology-PCR replacement strategy in at least two different backgrounds. Among these ORFs, two of them (YGL100w and YGL094c) are now known genes which encode well-characterized proteins (Seh1p, a nuclear pore protein, and Pan2p, a component of Pab1p stimulated poly(A) ribonuclease, respectively). The other four ORFs (YGL101w, YGL099w, YGL098w and YGL096w) code for proteins of unknown function, although the protein encoded by YGL101w has a strong similarity to the hypothetical protein Ybr242p. Gene disruptions were performed in diploid cells using the KanMX4 cassette, and the geneticin (G418)-resistant transformants were checked by PCR. Tetrad analysis of heterozygous deletant strains revealed that YGL098w is an essential gene for vegetative growth in three backgrounds, whereas the other five genes are non-essential, although we have found some phenotypes in one of them. YGL099wDelta strain did not grow at all at 15 degrees C and showed a highly impaired sporulation and a significantly lower mating efficiency. The other three deletants did not reveal any significant differences with respect to their parental strains in our basic phenotypic tests. PMID- 10669875 TI - Disruption of six novel genes from chromosome VII of Saccharomyces cerevisiae reveals one essential gene and one gene which affects the growth rate. AB - Six ORFs of unknown function located on chromosome VII of Saccharomyces cerevisiae were disrupted in two different genetic backgrounds, and the phenotype of the generated mutants was analysed. Disruptions of ORFs YGR256w, YGR272c, YGR273c, YGR275w and YGR276c were carried out using the disruption marker kanMX4 flanked by short homology regions, whereas ORF YGR255c was inactivated with a long flanking homology (LFH) disruption cassette (Wach et al., 1994). Tetrad analysis of the heterozygous disruptants revealed that ORF YGR255c, previously identified as COQ6 and encoding a protein involved in the biosynthesis of coenzime Q (Tzagoloff and Dieckmann, 1990), is an essential gene. The same analysis also revealed that sporulation of the ygr272cDelta heterozygous diploid produced two small colonies per ascus that were also G418-resistant, indicating that the inactivation of ORF YGR272c could result in a slower growth rate. This result was confirmed by growth tests of the haploid disruptants and by complementation of the phenotype after transformation with a plasmid carrying the cognate gene. No phenotypes could be associated to the inactivation of ORFs YGR256w, YGR273c, YGR275w and YGR276c. Two of these genes have recently been further characterized: ORF YGR255w, renamed RTT102, encodes a regulator of the Ty1-element transposition, whereas ORF YGR276c was found to encode the 70 kDa RNase H activity and was renamed RNH70 (Frank et al., 1999). PMID- 10669876 TI - Electrospray multistep ion trap mass spectrometry for the structural characterisation of poly AB - Electrospray 'soft' ionisation (ESI) and multistep mass spectrometry (MS(n)) techniques enable characterisation of a bioactive polymer, poly[(R,S)-3 hydroxybutanoic acid] (a-PHB), containing covalently bonded benzylpenicillin. The chemical structures of individual mass-selected bioactive macromolecules have been determined, and their fragmentation mechanisms have been compared with those of pure penicillin G. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669877 TI - Studies in organic mass spectrometry. Part 25. Benzyl ion formation in chemical ionisation (methane or isobutane) of some ortho-alkylhetero-substituted diphenylcarbinols AB - The behaviour of some ortho-alkylhetero-substituted diphenylcarbinols, including deuterium labelled derivatives, under chemical ionisation (methane or isobutane) conditions has been investigated. It has been determined that ortho-alkylhetero diphenylmethyl cations formed by water elimination from the protonated molecule undergo consecutive rearrangements which afford benzyl (or tropylium) ions previously observed for ortho-substituted diphenylcarbenium ions generated by electron ionisation. This reaction also occurs under low-energy collision conditions. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669878 TI - Validation of higher-throughput high-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry assays to conduct cytochrome P450s CYP2D6 and CYP3A4 enzyme inhibition studies in human liver microsomes. AB - In the early stage of drug discovery, thousands of new chemical entities (NCEs) may be screened before a single drug candidate can be identified for development. In order to accelerate the drug discovery process, we have developed higher throughput enzyme assays to evaluate the inhibition of cytochrome P450 isoforms 2D6 (CYP2D6) and 3A4 (CYP3A4) in human liver microsomes. The assays are based on high-performance liquid chromatography/tandem mass spectrometry (LC/MS/MS) techniques. The analysis time for each sample was reduced from approximately 20 minutes for the conventional HPLC assay to 30 seconds for the LC/MS/MS assay. For both LC/MS/MS assays, the linearity (r(2) > 0.99), precision (%CV < 15%) and accuracy (% bias <15%) for both inter- and intraday validations were satisfactory. Since the implementation of the LC/MS/MS assays, our sample throughput has increased by over 40-fold. PMID- 10669879 TI - Theoretical and numerical analysis of the behavior of ions injected into a quadrupole ion trap mass spectrometer AB - A numerical simulation method has been developed for the analysis of trapping ions injected into an ion trap mass spectrometer. This method was applied to clarify the effects of the following parameters on trapping efficiencies: (1) initial phase of the radio frequency (RF) drive voltage, (2) ion injection energy, and (3) RF peak voltage while injecting ions. The following conclusions were obtained by theoretical and simulation approaches. 1. The second and third dominant oscillations contribute significantly to the trapping mechanism of the injected ions, even for low q values. 2. A formula relating the operating parameters, which gives the maximum trapping efficiency, is derived. 3. Based on the above-mentioned formula, an advanced injection method is proposed, in which the RF peak voltage is decreased while injecting ions. The ability of this method to solve the problem of unequal sensitivity among different ion species is indicated by numerical simulation. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669880 TI - Determination of disulfide bonds in highly bridged alpha-dendrotoxin by matrix assisted laser desorption/ionization mass spectrometry AB - A methodology which combines tryptic digestions, 1,4-dithiothreitol reduction, and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) allows verification of the location of the disulfide bridges in alpha-dendrotoxin (C7-C57, C16-C40 and C32-C53). Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669881 TI - High throughput on-line solid phase extraction/tandem mass spectrometric determination of paclitaxel in human serum. AB - The feasibility of high throughput on-line solid phase extraction/tandem mass spectrometry (SPE/MS/MS) is tested for target analysis of paclitaxel in human serum. The use of a dual Prospekt system, with parallel SPE and elution directly to the mass spectrometer, resulted in a cycle time of 80 seconds for the entire, fully automated assay. The assay proved to be linear from 1 to 1000 ng/mL. Cartridges packed with small sorbent particles functioned both as SPE cartridges and as short analytical columns. PMID- 10669882 TI - Analysis of solid-supported oligonucleotides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry AB - This study presents matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) as a powerful tool to analyze and characterize oligonucleotides covalently linked to a solid support during their synthesis. The analysis of the fragment ions generated either in negative or positive mode allows direct and easy access to the nucleotide sequence and identification of the internucleosidic linkage. The mechanisms of the fragmentation of the solid supported oligonucleotides induced by MALDI-TOFMS are discussed. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669883 TI - Electrospray ionization mass spectrometry of synthetic oligonucleotides using 2 propanol and spermidine AB - Oligonucleotides have become widely used tools in molecular biology and molecular diagnostics. Their parallel synthesis in large numbers and the increasing interest in microarray technology has raised the requirement for fast and informative analytical tools for their quality control. A direct injection electrospray ionization mass spectrometry (ESI-MS) technique based on the use of aqueous 2-propanol as running eluent, and spermidine (or triethylamine) as DNA modifiers, has been applied to analyze a large set of samples (about 200 synthetic oligonucleotides) ranging from 5 to 15 kDa (17-51mers) with good results in terms of sensitivity, suppression of sodium adduct formation, and speed of analysis. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669884 TI - Ion trap MS(n) genealogical mapping-approaches for structure elucidation of novel products of consecutive fragmentations of morphinans AB - The analysis of data within multi-generational, genealogical, electrospray ionization/MS(n) fragmentation maps is discussed in reference to the structure elucidation of morphinans, an important class of pharmacological compounds. Various general approaches to separate and understand observed processes are discussed. These include: (1) Simple synthetic schemes incorporating deuterium and (13)C into morphinans to study later-generation, O-methyl group migration; (2) labeling to understand 'intense' signals for even-electron to odd-electron ion 'switching' events; (3) gas phase 'synthesis' of proposed MS(3) ions via an independent route, using chemical ionization (CI) MS/MS of naphthalenes and analysis via a bench-top El/Cl ion trap; (4) a useful synthetic paradigm for generating proposed carbonium ion structures at MS(4) via electrospray ionization (ESI) of easily synthesized amines; (5) the analysis of an oxidation product and correlation of MS(n) data of a switched, odd-electron species with electron ionization and low-pressure, low-energy charge exchange data; and (6) a new way of summarizing MS(n) data. Copyright -Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669886 TI - Origin of mass shifts in the quadrupole ion trap: dissociation of fragile ions observed with a hybrid ion trap/mass filter instrument AB - A novel hybrid tandem mass analyzer, coupling a quadrupole ion trap with a quadrupole mass filter, has been constructed to permit mass analysis of ions ejected from the ion trap. The initial application of this instrument is the investigation of the origin of mass shifts in the ion trap due to ion fragility. We hypothesize that fragile ions undergo mass shifts, characterized by peak fronting, due to early ejection from the quadrupole ion trap. As these ions come into resonance with the ejection frequency, they gain kinetic energy, collide with buffer gas molecules and thus can dissociate to produce fragment ions. These fragment ions will not be stable within the ion trap as they are situated past the stability boundary at q(z) = 0. 908. Consequently the fragment ions are ejected prematurely. This results in an apparent mass shift due to peak fronting. The experiments reported here clearly document the production of fragment ions as the origin of mass shifts during the resonant ejection of fragile ions. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669885 TI - Elucidation of peptide metabolism by on-line immunoaffinity liquid chromatography mass spectrometry. AB - An immunoaffinity chromatography extraction capillary liquid chromatography separation has been coupled to electrospray ionization mass spectrometry for on line characterization of drug metabolites of a therapeutic peptide in plasma. It is demonstrated that the selectivity, sensitivity and molecular weight data provided by immunoaffinity chromatography coupled to liquid chromatography/mass spectrometry provides a means of rapidly achieving qualitative determinations of small amounts of material in complicated biological matrices such as plasma. The ability to detect the peptide in rat plasma at a level of 10 ng/mL is demonstrated using this method. In addition, experiments to study the epitope of the peptide by enzymatic digestion and mass spectrometry are also discussed. The method is proposed as an alternative approach to studying the metabolism of therapeutic peptides. PMID- 10669887 TI - Frozen shell approximation violation in negative ion formation from halogenated benzenes via dissociative attachment AB - A series of benzene derivatives (R(1)C(6)H(4)R(2)) has been studied by means of electron capture negative ion mass spectrometry (ECNI-MS), and PM3 quantum chemical calculations. The dissociation channel M(-.) --> Hal(-) + (M - Hal). is analysed from the point of view of symmetry conservation. Generally, a symmetry ban on dissociation may be avoided in at least two ways: (i) out-of-plane vibrations of the halogen atom in the molecular negative ion (MNI), mixing pi- and sigma-states of the anion; (ii) symmetrical in-plane vibration of the C-Hal bond, changing the order of the empty levels in the MNI with subsequent radiationless conversion into a sigma-state. Our analysis shows that neither of them provides a satisfactory explanation of the ECNI mass spectra for chlorobenzene, if one retains the usual assumption that an additional electron goes into the LUMO of the neutral molecule. Thus, it may be concluded that in this case electron capture causes a significant perturbation of the energy ordering of vacant orbitals, thus making the frozen shell approximation inapplicable. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669888 TI - Determination of tetracyclines in bovine kidney by liquid chromatography/tandem mass spectrometry with on-line extraction and clean-up. AB - A novel, sensitive, high performance liquid chromatography/tandem mass spectrometric (i.e. mass spectrometry/mass spectrometry) method with on-line extraction and clean-up for the screening and confirmation of residues of tetracyclines in kidney has been developed. After liquid extraction of homogenised kidney with McIlvain buffer, an aliquot of the extract is directly injected on the LC/MS/MS system with further extraction and clean-up of the sample on-line. Detection of the analytes was achieved by positive electrospray ionization followed by multiple reaction monitoring. For each tetracycline the collisional decomposition of the protonated molecule to a unique, abundant fragment ion was monitored. The method has been validated for tetracycline, oxytetracycline, chlortetracycline and doxycycline. Calibration curves resulting from spiked blank kidney samples at the 100-1200 microgram/kg level showed good linear correlation. At the level of 600 microgram/kg both within- and between-day precision, as measured by relative standard deviation (RSD), were less than 7%. The limits of detection (LODs) for tetracycline, oxytetracycline, chlortetracycline and doxycyline were 18, 23, 24 and 21 microgram/kg, respectively. The limits of quantification (LOQs) for tetracycline, oxytetracycline, chlortetracycline and doxycyline were 36, 46, 47 and 42 microgram/kg, respectively. The recoveries ranged from 71 to 91%. The procedure provides a rapid, reliable and sensitive method for the determination of residues of tetracyclines in bovine kidney. The advantage of this method over existing methods is its decreased sample preparation and analysis time, which makes the method more suitable for routine analysis. PMID- 10669889 TI - On-line size-exclusion chromatography/electrospray ionisation mass spectrometry of aquatic humic and fulvic acids AB - Isolated aquatic humic and fulvic acids were analysed with on-line size exclusion chromatography/electrospray ionisation mass spectrometry (SEC/ESI-MS). An eluent composition which enabled electrospray ionisation was identified. The SEC separation improved interpretability of mass spectra and may open up new possibilities for molecular weight determination of humic substances. A linear dose-response relationship over a factor of 20 was obtained and the limit of detection was 50ng/uL for humic and fulvic acids. Spectral changes due to different ionisation conditions (pH and cone voltage) were investigated. A natural water sample from a Swedish lake was analysed. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669890 TI - Sample vial influences on the accuracy and precision of carbon and oxygen isotope ratio analysis in continuous flow mass spectrometric applications AB - Due to the small amounts of sample gas involved in continuous flow mass spectrometric analysis, care should be taken to evaluate the influence of sample containers on the carbon and oxygen isotope ratios of samples. Data indicate that Na-glass and borosilicate glass vials, equipped with butyl rubber septa, can cause significant changes in the isotopic composition of CO(2) gas, even where sample gases are stored within the vial for less than one day. The magnitude of these changes varies from vial to vial. Given the leverage that contamination can potentially exert on small gas samples, each researcher should carefully evaluate the effect of sample vials in order to eliminate unknown and unwanted changes in the composition of samples. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669891 TI - Characterization of organometallic coordinative cluster compounds of silver by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was successfully applied to characterize the organosilver coordinative cluster compounds, silver phenylacetylide and three silver thiolates, namely, silver tertiary butylthiolate, silver 2,6 dimethylbenzenethiolate, and silver 2, 6-dichlorobenzenethiolate. Samples and dithranol matrix were finely dispersed in 1:1 tetrahydrofuran (THF)/chloroform (CHCl(3)) mixed solvent. In most cases the monomer units remained intact during ionization, and the oligomeric molecular ions were produced through silver cationization, with a general molecular ion formula [nM + Ag](+). This was further verified by the relative abundances of the isotopic peaks within the molecular ion clusters, which were in close agreement with those theoretically calculated for nM cationized with one silver ion. In the case of silver 2, 6 dichlorobenzenethiolate, in addition to the dominant [nM + Ag](+) peaks, weak peaks corresponding to the successive losses of hydrogen chloride molecules were observed. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10669892 TI - Density-dependent dynamics and superinfection in an epidemic model. AB - A mathematical model of the interaction between two pathogen strains and a single host population is studied. Variable population size, density-dependent mortality, disease-related deaths (virulence), and superinfection are incorporated into the model. Results indicate that coexistence of the two strains is possible depending on the magnitude of superinfection. Global asymptotic stability of the steady-state that gives coexistence for both strains under suitable and biologically feasible constraints is proved. PMID- 10669893 TI - A Kermack-McKendrick model applied to an infectious disease in a natural population. AB - The dynamics of a fatal infectious disease in a population regulated by density dependent constraints are represented as a system of nonlinear integral equations. Survival probabilities and disease transmission coefficients may vary with the time elapsed since infection, and horizontal and vertical modes of transmission are allowed for. Criteria for the existence and stability of steady states are derived, and an example based on the dynamics of tuberculosis is presented. Finally, the relative merits of this approach, and the familiar compartmental models based on differential equations are discussed. PMID- 10669894 TI - Multihost, multiparasite systems: an application of bifurcation theory. AB - The local analysis of multihost multiparasite models has been hampered by algebraic intractability. There have been two responses to this difficulty: extensive numerical investigation, and simplification to a level where analytical techniques work. In this paper we describe another approach, based on bifurcation theory, in which the qualitative properties of the model equilibrium structure are realized on an array of maps drawn in parameter space. This approach is described in the context of two models: the basic two-host shared microparasite S I model and the single-host two-microparasite S-I (susceptible-infective) model. The procedure involved does not require model simplification through a reduction in dimensionality. It can handle intraspecific as well as parasite-mediated competition and, in the second model, single-host parasite coexistence. The map arrays provide a concise catalogue of the possible modes of behaviour of a system and an explanation for changes in that behaviour. In particular, the reasons why the conjectures made about the behaviour of the first of these models do not hold throughout parameter space are immediately clear from the map structure, as are the conditions for collusive and competitive behaviour between the two types of parasite in the second model. PMID- 10669895 TI - Modelling corneal epithelial wound closure in the presence of physiological electric fields via a moving boundary formalism. AB - A new framework for the modelling of corneal epithelial wound healing is presented, which can include the presence of a physiological electric field. The difficulty inherent in the inclusion of this biological phenomenon motivates our use of a moving boundary formalism. A key conclusion is that the model predicts a linear relation between the wound healing speed and the physiological electric field strengths over a physiologically large range of electric field strength. Another key point is that this linear relationship between electric field strength and wound healing speed is robust to variations in critical parameters that are difficult to estimate. The linearity is also robust to different realizations of the modelling framework presented. PMID- 10669896 TI - On a branching model of division-within-division. AB - We consider a deterministic version of a stochastic model for division-within division processes described by Kimmel (1997, In: Proceedings of the IMA Workshop 'Classical and Modern Branching Processes' (K. Arthreya and P. Jagers, eds.)???? :????). It is shown that the behaviour of the deterministic model can be analyzed by using an associated Markov chain, using the methods of Barbour et al. PMID- 10669897 TI - Pharmacokinetics of benflumetol given as a fixed combination artemether benflumetol (CGP 56697) in Thai patients with uncomplicated falciparum malaria. AB - The pharmacokinetics of benflumetol as a fixed combination, artemether benflumetol (CGP 56697), following three regimens [regimen A: four tablets at 0, 8, 24 and 48 h (320 mg artemether, 1,920 mg benflumetol); regimen B: two tablets at 0, 8, 24 and 48 h (160 mg artemether, 960 mg benflumetol); regimen C: four tablets at 0, 8 and 24 h (240 mg artemether, 1,440 mg benflumetol)] were investigated in 39 patients with acute uncomplicated falciparum malaria. All patients showed a rapid initial response with a median parasite clearance time of 40, 41 and 39.5 h and a fever clearance time of 27.8, 32 and 24.5 h for regimens A, B and C, respectively. In nine patients (two, four and three patients in regimens A, B and C, respectively), however, parasitemia reappeared in the peripheral blood smear between days 9 and 23. The pharmacokinetics of benflumetol were highly variable, with coefficients of variation in pharmacokinetic parameters ranging from 14.9% to 144%. Absorption and elimination of benflumetol were relatively slow. Median Cmax per dose (first dose) was significantly higher in regimen B (6.29 ng/ml/mg dose) than in regimen A (2.6 ng/ml/mg dose) and regimen C (3.06 ng/ml/mg dose). Mean T1/2z in regimen C (2.65 h) was significantly shorter than in regimen A (4.5 h) and regimen B (3.89 h). In patients on regimens A and B who showed a sensitive response, plasma concentrations of benflumetol were significantly higher than in those with treatment failure. PMID- 10669898 TI - Analgesic effects of diclofenac suppository and injection after preoperative administration. AB - Diclofenac sodium (100 mg) has been introduced in the Caribbean as a suppository formulation. In a randomized single-blind (observer-blind) clinical trial, the postoperative analgesic efficacy of diclofenac administered either as a conventional intramuscular injection (75 mg) or as the available suppository formulation (100 mg) was studied in 44 adult male patients undergoing herniorrhaphy in same day surgery. Diclofenac was administered preoperatively at induction of anesthesia to patients (grades ASA I and II) after they had given informed consent. Evaluation of analgesia on the visual analog scale (VAS) did not differ significantly between the two treated groups at three assessment times: on admission to the recovery room, the postoperative ward and at discharge. The times for requests for additional analgesia and the number of patients requesting further analgesia did not differ. Patients who received the suppository were discharged earlier than those who received the injection (40 min vs. 65 min p = 0.02). This preliminary study of the two marketed formulations of diclofenac demonstrated that both preparations provided equivalent analgesia but patients who received the suppository preparation were discharged earlier. PMID- 10669899 TI - Effect of a mineral water on gastric emptying of patients with idiopathic dyspepsia. AB - The antidyspeptic property of mineral waters has for many years been based on empirical data. In the present paper we evaluated the effects of one type of mineral water, Tettuccio water from Montecatini, on gastric emptying in patients with idiopathic dyspepsia. Fourteen subjects, eight patients with idiopathic dyspepsia and delayed gastric emptying at scintigraphy and six healthy subjects with normal gastric emptying were studied. The gastric emptying of mineral water was studied with a scintigraphic method and compared with that of tap water. In patients with idiopathic dyspepsia, gastric emptying of both waters was slower than in controls but the gastric emptying of mineral water was significantly faster than that of tap water, both in dyspeptic patients and in healthy subjects. In conclusion, this mineral water stimulates gastric emptying. Further studies are needed on the possible role of this water in the management of chronic idiopathic dyspepsia. PMID- 10669900 TI - Efficacy and safety of sumatriptan 50 mg in patients not responding to standard care, in the treatment of mild to moderate migraine. The Sumatriptan 50 mg Italian Study Group. AB - The tolerability and efficacy of oral sumatriptan 50 mg for the treatment of mild to moderate migraine attacks were assessed in a double-blind, multicenter placebo controlled study on a group of patients who had not responded sufficiently to analgesic preparations. Three-hundred-and-twenty-eight migraine sufferers treated a first migraine attack with a nontriptan standard care medication: a mixture containing phenazone, butalbital and caffeine (optalidon) or indomethacin plus prochlorperazine plus caffeine (difmetre) or paracetamol 100 mg (tachipirine), depending on their habits. Of these patients, 32.6% reported headache relief with this treatment and were not included in phase II of the study. The 219 patients not reporting relief during the first phase of the study entered the second phase and were randomized to sumatriptan 50 mg or to placebo; 167 of these patients treated a second attack according to the protocol and were evaluated for efficacy. Of the patients with migraine taking sumatriptan, 58% reported headache relief compared with 35% of placebo-treated patients (p = 0.008). The reduction of nausea and vomiting was significantly better in the sumatriptan group. No differences were detected for the recurrence rate, while rescue medication was used more by the placebo group. The safety profile of sumatriptan 50 mg was confirmed. This study demonstrates the usefulness of this dose of oral sumatriptan against the pain and the accompanying symptoms of mild and moderate migraine. PMID- 10669901 TI - Proliferation of spleen cells in culture stimulated by 7-thia-8-oxoguanosine: evidence that both B- and T-cells are the targets of its action. AB - 7-thia-8-oxoguanosine (immunosine) is a nucleoside analog showing efficient antiviral activity in rodent models as a consequence of enhancement of the immune response. However, little is known about the mechanisms of its action. In this work the effect of immunosine on proliferation of mouse and rat splenocytes in culture was studied. It was found that the compound stimulated proliferation of lymphocytes in a dose-dependent manner without any additional stimuli. The effect is predominantly mediated by interleukin-2 (IL-2) as judged by increased IL-2 production, upregulation of IL-2 receptor alpha (IL-2R alpha) expression and by significant inhibition (60-75%) of cell proliferation by anti-IL-2R alpha monoclonal antibodies (mAbs). Immunosine also stimulated proliferation both of T- and B-cells purified by immunomagnetic sorting. The response of B-cells was much higher than that of T-cells. The stimulatory effect of immunosine on both lymphocyte subpopulations was further increased by the addition of enriched splenic antigen-presenting cells or purified dendritic cells. Proliferation of purified T-cells to immunosine was also significantly potentiated by an anti alpha beta T-cell receptor mAb (R 73). All these data suggest that T-, B- and accessory cells in splenic cultures are the targets of immunosine action. PMID- 10669902 TI - Effects of newly synthesized analogs of MIF-1 containing unnatural amino acids on electrically evoked smooth muscle contractions. AB - New MIF-1 (Pro-Leu-Gly-NH2) analogs containing unnatural amino acids such as L canavanine (Cav) and L-cysteic acid S-(2-aminoethyl)amide (sLys) have been synthesized and in vitro experiments were performed to study their action on neurotransmission in target tissues with adrenergic and cholinergic neurotransmission. The experiments were carried out on electrically stimulated proximal guinea pig ileum (GPI) and the prostatic part of rat and rabbit vasa deferentia (VDR, VDRabb). The present results show that the newly synthesized Cav2-MIF and sLys2-MIF might affect electrically evoked smooth muscle contractions. PMID- 10669903 TI - Involvement of spinal delta 1-opioid receptors in forced walking stress-induced antinociception in the tail-flick test in mice. AB - The purpose of this study was to elucidate the involvement of spinal delta-opioid receptor subtypes in forced walking stress-induced antinociception mice. We first confirmed that forced walking stress produced walking duration-dependent antinociception in mice as determined by the tail-flick test. Intrathecal treatment with 7-benzylidenenaltrexone, a selective delta 1-opioid receptor antagonist, significantly attenuated forced walking stress-induced antinociception. In contrast, intrathecal treatment with naltriben, a selective delta 2-opioid receptor antagonist, had no significant effect on forced walking stress-induced antinociception. Intracerebroventricular treatment with either 7 benzylidenenaltrexone or naltriben had no effect on the forced walking stress induced antinociception. These results suggest that forced walking stress-induced antinociception is mediated by spinal delta 1-opioid receptors in mice. PMID- 10669904 TI - Effects of repeated selective serotonin reuptake inhibitor paroxetine treatments on mouse forced swimming. AB - Studies were performed in the mouse forced swimming model, a well known experimental depression model, in order to detect the mechanism of the antidepressive effects induced by repeated serotonin reuptake inhibitor (SSRI) dosing. Five-day repeat dosing of a typical SSRI, paroxetine, increased climbing, a distinctive antidepressive behavior, 1 h after but not 1 h before treatment. The coinjection of paroxetine and serum in mice treated with four repeated doses of paroxetine distinctively increased the behavior, but the coinjection of paroxetine and serum in mice without paroxetine did not. These results indicate that repeated dosing of paroxetine produces a serum substance related to the antidepressive effects induced by serotonin neuron activities. Furthermore, the behavior induced by 5-day repeated dosing of paroxetine was decreased by 100 and 10 micrograms/kg of ketanserin (5-HT2 antagonist) and 100 micrograms/kg of LY 278584 (5-HT3 antagonist). The present findings strongly suggest that repeated dosing of paroxetine produces a serum substance stimulating the antidepressive neuronal pathway sensitively mediated by 5-HT2 and 5-HT3 receptor activity. PMID- 10669905 TI - Quinpirole, 8-OH-DPAT and ketanserin modulate catalepsy induced by high doses of atypical antipsychotics. AB - The effect of the selective dopamine D2 receptor agonist quinpirole, the selective 5-HT1A receptor agonist 8-OH-DPAT and the selective 5-HT2A receptor antagonist ketanserin on catalepsy induced by atypical antipsychotics clozapine, risperidone, olanzapine and sertindole at higher doses was studied in rats. Haloperidol (0.5, 1 and 2 mg/kg), clozapine (50 and 75 mg/kg) and olanzapine (15 and 30 mg/kg) produced catalepsy dose-dependently while sertindole at doses up to 40 mg/kg failed to produce catalepsy in rats. However, sertindole (15, 30 and 45 mg/kg) produced a cataleptic effect in mice in a dose-dependent manner. At a high dose (5 mg/kg), risperidone produced catalepsy in rats. Quinpirole (0.05 and 0.1 mg/kg) reversed the cataleptic effect of haloperidol (2 mg/kg), risperidone (5 mg/kg), olanzapine (30 mg/kg) and sertindole (45 mg/kg). Quinpirole (0.05 and 0.1 mg/kg) reversed clozapine (75 mg/kg)-induced catalepsy. 8-OH-DPAT (0.15 and 0.3 mg/kg) dose-dependently reversed catalepsy induced by haloperidol (2 mg/kg) and risperidone (5 mg/kg) without affecting the cataleptic effect of olanzapine. However, the higher dose (0.45 mg/kg) of 8-OH-DPAT reversed it significantly. 8 OH-DPAT (0.3 mg/kg) reversed clozapine (75 mg/kg)-induced catalepsy. 8-OH-DPAT (0.15, 0.3 and 0.45 mg/kg) failed to reverse sertindole-induced catalepsy. Ketanserin (0.75 and 1.5 mg/kg) completely reversed catalepsy induced by haloperidol (2 mg/kg) and risperidone (5 mg/kg). Ketanserin (0.75 and 1.5 mg/kg) dose-dependently reversed olanzapine (30 mg/kg) and sertindole (45 mg/kg)-induced catalepsy without any effect on clozapine (75 mg/kg)-induced catalepsy. A higher dose (3 mg/kg) of ketanserin reversed clozapine-induced catalepsy. The present study suggests that atypical antipsychotics show fewer extrapyramidal symptoms (EPS) due to greater modulation of the serotonergic system. Therefore, an antipsychotic with dopamine D2/5-HT2A antagonistic action and 5-HT1A agonistic action may prove to be superior to the existing antipsychotics. PMID- 10669906 TI - Role of nitric oxide in electroshock and pentylenetetrazole seizure threshold in rats. AB - There are contradictory reports on whether nitric oxide (NO) is a proconvulsant or anticonvulsant. Hence a study was designed to investigate the effect of NO donor l-Arginine and NO synthesis inhibitor N omega-nitro-L-arginine (NOARG) on electroshock- and pentylenetetrazole (PTZ)-induced seizure threshold in rats. L arginine was tested in three doses (75, 150 and 300 mg/kg), and NOARG was administered in doses of 4, 8 and 16 mg/kg. L-Arginine increased the intensity of current required to produce a threshold seizure, whereas NOARG had the opposite effect. In PTZ-induced seizures, L-arginine significantly decreased the dose of PTZ required to produce a threshold seizure, while NOARG increased it. Hence, it was concluded that NO synthase inhibition had the opposite effect in electroshock and PTZ-induced seizures, meriting further studies on the mechanism of effect. PMID- 10669907 TI - Effects of histamine and related compounds on regional cerebral blood flow in rats. AB - The effects of histamine and related compounds on regional cerebral blood flow (rCBF) in the hippocampus of conscious rats were studied. Intracerebroventricular injection of histamine caused a dose-dependent increase in rCBF in the hippocampus, and similar findings were observed with not only the H1 agonist, 2 thiazolylethylamine, but also the H2 agonist, dimaprit. Intraperitoneal injection of L-histidine also resulted in an increase in rCBF in the hippocampus, in parallel with elevation of histamine content in the brain. The increase in rCBF in the hippocampus induced by L-histidine was antagonized by both H1 and H2 antagonists (diphenhydramine, pyrilamine and zolantidine). In addition, when both antagonists were injected simultaneously, an additive effect was observed in antagonism of the L-histidine-induced increase in rCBF. L-Histidine caused no marked changes in blood pressure even at a dose of 1,500 mg/kg, which showed an increase in rCBF in the hippocampus. These results indicate that histamine elicited an increase in rCBF via both H1 and H2 receptors. PMID- 10669908 TI - Enhanced cardiovascular reactivity to desmopressin in water-restricted rats: facilitatory role of immunosuppression. AB - There is considerable evidence suggesting that vasopressin may play an important role in regulating cardiovascular homeostasis. In the present study the effects of immunosuppressant drugs, ciclosporin and tacrolimus on arterial blood pressure (BP) and heart rate (HR) responses to desmopressin (DDAVP) were compared in anesthetized normally hydrated and 24-h water-restricted rats. When injected intravenously, single bolus doses of DDAVP (0.5-8.0 micrograms/kg) elicited dose dependent decreases in BP and also attenuated the HR. The enhancement of the vasodepressor response to DDAVP (4 micrograms/kg) was produced 15 min after the intravenous injection of either ciclosporin (3 mg/kg) or tacrolimus (25 micrograms/kg) accompanied by reduction in HR in water-restricted animals, but the response in euhydrated animals remained less modified after the bolus injections of immunosuppressants and DDAVP. To elucidate the possible contribution of the renal vascular system in water-restricted rats, ciclosporin was administered intravenously, followed 15 min later by 4 micrograms/kg injections of arginine vasopressin (AVP) or DDAVP into the renal artery, but there was no significant change in both the cardiovascular parameters as compared with the responses obtained with agonists alone. The present results indicate that the augmentation of the cardiovascular response to DDAVP by ciclosporin and tacrolimus in water-restricted anesthetized rats appears to be mediated by the release of endogenous vasodilatory mediators. PMID- 10669909 TI - Bioequivalence study of rifampicin in fixed-dose combination of rifampicin and isoniazid vs. separate formulations. AB - The benefits of fixed-dose combinations of antituberculosis agents are well recognized by the World Health Organization (WHO) and International Union Against Tuberculosis and Lung Disease (IUATLD) and preferred over separate formulations. Therefore, a comparative bioequivalence study of rifampicin and isoniazid together in a fixed-dose combination and separately (at the same dose levels) was performed on a group of 12 healthy subjects. The study was designed as a single blind, crossover experiment. Nine blood samples were collected from each subject over a period of 24 h. The plasma concentrations of rifampicin were assessed by a method developed in this laboratory. Various pharmacokinetic parameters of rifampicin such as AUC, Cmax, Tmax and t1/2 were also calculated. The study demonstrates that a fixed-dose combination (test formulation) and separate formulations (standard formulations) are bioequivalent for rifampicin. PMID- 10669910 TI - Lack of pharmacokinetic interaction between grapefruit juice and phenytoin in healthy male volunteers and epileptic patients. AB - The present study was undertaken to determine the effects of grapefruit juice (GFJ) on the oral bioavailability of phenytoin (DPH). Ten healthy male volunteers received a single dose of 300 mg of DPH orally with 300 ml of water or fresh GFJ in a randomized crossover fashion. In both instances, blood sampling was done at 0, 0.5, 1, 2, 4, 6, 9, 12, 24, 48, 72, 96 and 120 h. The study was also carried out in 10 epileptic patients with uncontrolled seizures even after 3 to 4 weeks regular therapy of DPH (100 mg three times a day). Blood samples were drawn at 0, 1, 2, 3, 4, 5, 6 and 8 h after DPH with 300 ml of water or fresh GFJ. No significant difference was observed in any of the pharmacokinetic parameters like Cmax, Cmin, Tmax, t1/2a, t1/2e and AUC of DPH after GFJ administration as compared to control values in healthy volunteers and epileptic patients. It is concluded that GFJ does not affect the oral bioavailability of DPH. PMID- 10669911 TI - Double-blind placebo-controlled study with citicoline in APOE genotyped Alzheimer's disease patients. Effects on cognitive performance, brain bioelectrical activity and cerebral perfusion. AB - Cytidine 5'-diphosphocholine (citicoline) is a an endogenous intermediate in the biosynthesis of structural membrane phospholipids and brain acetylcholine. Citicoline has been extensively used for the treatment of neurodegenerative disorders associated with head trauma, stroke, brain aging, cerebrovascular pathology and Alzheimer's disease. In this study we have investigated the efficacy and safety of the treatment with citicoline versus placebo in patients with Alzheimer disease. Thirty patients (age = 73.0 +/- 8.5 years; range = 57-87 years) with mild to moderate senile dementia (GDS: stages 3-6) of the Alzheimer type were included in a double-blind, randomized and placebo-controlled clinical trial. After a 2-week period of drug washout, patients were treated with i) placebo (n = 17; age = 73 +/- 5 years) or ii) 1,000 mg/day of citicoline (n = 13; age = 76 +/- 9 years) for 12 weeks (84 days). Examinations were done at baseline (T0) and after the 12 weeks of treatment (T12). As compared to placebo, citicoline improved cognitive performance in Alzheimer's disease patients with APOE E4 (ADAS: difference between groups = -3.2 +/- 1.8 scores, p < 0.05; ADAS cog: difference between groups = -2.3 +/- 1.5, ns); and this improvement on cognition was more pronounced (ADAS, p < 0.01; ADAS-cog: difference between groups = -2.8 +/- 1.3, p < 0.06) in patients with mild dementia (GDS < 5). Citicoline also increased cerebral blood flow velocities in comparison with placebo (p < 0.05) when transcranial Doppler recordings from both hemispheres were considered together, as well as diastolic velocity in the left middle cerebral artery (p < 0.05). Patients treated with citicoline showed an increase in the percentage of brain bioelectrical activity of alpha (occipital electrodes) and theta type (left side electrodes), accompanied by a decrease in relative delta activity particularly marked in the left temporal lobe. Significant differences with respect to placebo (p < 0.05) were observed for theta activity in several fronto-parieto-temporal electrodes of the left hemisphere. Treatment with citicoline tended to reduce serum IL-1 beta levels, mainly after 4 weeks of administration, with no modified blood histamine content. In addition, neither adverse side effects nor alterations in biological and hematological parameters were induced by citicoline. The present data indicate that citicoline (1,000 mg/day) is well tolerated and improves cognitive performance, cerebral blood perfusion and the brain bioelectrical activity pattern in AD patients. According to our results, it seems that citicoline might be a useful treatment in Alzheimer's disease, and that the efficacy of this compound is greater in patients with mild mental deterioration and/or bearing the epsilon 4 allele of the APOE. PMID- 10669912 TI - Porcine insulin biodegradable polyester microspheres: stability and in vitro release characteristics. AB - The stability of porcine insulin in biodegradable polymers, i.e., poly(DL-lactide co-glycolide) 50:50 (50:50 DL-PLGA) and poly(L-lactide) (L-PLA) was investigated. Insulin encapsulated microspheres were fabricated from both polymers using double emulsion-solvent evaporation and emulsion-solvent evaporation techniques and subjected to accelerated stability studies at 40 degrees C and 75% relative humidity. Porcine insulin was found to degrade in all microsphere formulations with an average of < 50% of the initial loading amount remaining intact at the end of 4 weeks. The two major degradation products observed in these formulations were determined to be A-21 desamido insulin and covalent insulin dimer with trace amounts of high molecular weight transformation products. In vitro release studies in phosphate buffered saline at 37 degrees C resulted in very slow and incomplete (< 30% in 30 days) release kinetics for all microsphere formulations. Extraction and analyses of the unreleased insulin within the microspheres revealed that an average of approximately 11% of the encapsulated insulin remained intact. The degradation products observed consisted of approximately 15% of three distinct deamidated hydrolysis products including A-21 desamido insulin, approximately 22% covalent insulin dimer, and trace amounts of high molecular weight transformation products. The degradation of porcine insulin within biodegradable polyester microspheres during stability and release studies can be attributed to the gradual decrease in the pH within the microspheres due to progressive polymer hydrolysis resulting in the production of DL-lactic and glycolic acids. The encapsulation of an acid-base indicator, bromophenol blue, in 50:50 PLGA microspheres (as a probe to estimate pH within the microspheres during accelerated stability studies) indicated that the pH decreased to approximately 3.8 after 3 weeks. PMID- 10669913 TI - An evaluation of fluid bed drying of aqueous granulations. AB - The purpose of the work described was twofold: (a) to apply heat and mass balance approaches to evaluate the fluid bed drying cycle of an aqueous granulation, and (b) to determine the effect of the temperature and relative humidity of the drying air on the ability to meet a predetermined moisture content specification. Water content determinations were performed using Karl Fischer titration, and Computrac and Mark 1 moisture analyzers. The water vapor sorption isotherms were measured using a gravimetric moisture sorption apparatus with vacuum-drying capability. Temperature, relative humidity, and air flow were measured during the drying cycle of a production-scale fluid bed dryer. Heat and mass balance equations were used to calculate the evaporation rates. Evaporation rates calculated from heat and mass balance equations agreed well with the experimental data, whereas equilibrium moisture content values provided useful information for determination of the upper limit for inlet air humidity. Increasing the air flow rate and inlet temperature reduced the drying time through the effect on the primary driving force. As expected, additional drying of granules during the equilibration period did not show a significant impact on reducing the final moisture content of granules. Reducing the drying temperature resulted in measurement of higher equilibrium moisture content for the granules, which was in good agreement with the water vapor sorption data. Heat and mass balance equations can be used to successfully model the fluid bed drying cycle of aqueous granulations. The water vapor sorption characteristics of granules dictate the final moisture content at a given temperature and relative humidity. PMID- 10669914 TI - Effects of the metal type and the roughness of the die wall on the expended work for tablet ejection. AB - The aim of the present work was to obtain an impression about the consequences for the tableting process when dies of different quality are used. Two hard metal dies and one die made from tool steel, each with a distinct roughness of the die wall, were compared by compacting dry-blended powder mixtures on an eccentric tablet press. The feasibility of the tableting process was assessed by quantifying the ejection force and the work expended by the lower punch during a first phase of the ejection process. With decreasing amounts of lubricant, abrupt permanent impacts were observed leading to conditions at which an ejection was no longer possible. This observation did not depend on the roughness of the die wall used or on the compaction pressure, but strongly depended on the metal type of the die used and on the tested formulation. For the tool steel no difference was found in this respect between the two tested formulations (0.5% versus 2% silica aerogel). In either case, a concentration of 0.3% magnesium stearate was sufficient; however, a concentration of 0.2% magnesium stearate was not sufficient. For both hard metal dies, concentrations of 0.3% (0.5% silica aerogel) and 0.5% magnesium stearate (2% silica aerogel) were definitely not sufficient. Within the ranges above a minimum lubrication, the ejection force and the ejection work increased with the degree of the die wall roughness on a scale comparable to that of the tested formulation factors. In particular, if changes of the die tooling are likely to occur in the life cycle of a product, it is highly recommended that the quality of the dies be considered in the development phase. PMID- 10669915 TI - Influence of crystal habit on the surface free energy and interparticulate bonding of L-lysine monohydrochloride dihydrate. AB - The objective of the present study was to apply a technique to measure the surface energy of crystalline powders without changing the surface properties by compaction, and to relate such measurements to crystal habit and orientation. The surface free energy of uncompacted L-lysine monohydrochloride dihydrate (LH), determined using a modified sessile-drop method, reflected a combined value for the various faces, and was influenced by the relative size of the faces and the orientation of the crystals. The surface free energy values obtained from contact angle measurements were within the possible range calculated from the crystal structure. Discrepancies between the theoretical estimates of interparticulate cohesive strengths and those measured from the tensile strength of powder compacts were used to estimate the flaw sizes (or gaps between the particles) that act as stress concentrators and reduce the tensile strength of the compacts. The flaw sizes indicate packing and compressibility of the various crystal habits. In the absence of compressive load, compacts made out of the equidimensional crystals have the larger flaw sizes (wider cracks or wider gaps between the particles). At higher compaction pressures, the compacts from long rod-shaped crystals have longer crack lengths. The weakness of the compacts made from the long rods at the higher compaction pressures may be because of the longer crack length along the interparticulate boundary, which may result in a higher stress intensity at the crack tip and increased fracture propensity. PMID- 10669916 TI - Formation and stability of the dispersed particles composed of retinyl palmitate and phosphatidylcholine. AB - The purpose of this study was to develop an intravenous formulation composed of retinyl palmitate (RP) for the treatment of cancer. RP was dispersed with soybean phosphatidylcholine (PC) using sonication and the dispersal mechanism was evaluated by characterizing the dispersed particles using dynamic light scattering, fluorescence spectroscopy, and surface monolayer techniques. The dispersions in the RP mole fraction range of 0.1-0.8 were stable at room temperature for 3 days. A limited amount of RP was incorporated into PC bilayer membranes (approximately 3 mol%). The excess RP separated from the PC bilayers was stabilized as emulsion particles by the PC surface monolayer. When the PC content was less than the solubility in RP, the PC monolayer did not completely cover the hydrophobic RP particle surfaces and separation into oil/water occurred. The miscibility between RP and PC and the lipid composition were critically important for the stability of the dispersed particles (coexistence of emulsion particles [surface monolayer of PC + core of RP] with vesicular particles [bilayer]) of the lipid mixtures. PMID- 10669917 TI - Characteristics of pharmaceutical grade phyllosilicate powders. AB - The purpose of this paper was to evaluate the possible use of three types of pharmaceutical grades of phyllosilicates as pharmaceutical excipients. Seven samples (two smectites, three palygorskites, and two sepiolites) were studied. A complete mineralogical and chemical characterization of each material was made and powder texture was established by image analysis of scanning electron micrographs. Powder flow properties were then determined and the influence of textural characteristics on powder rheology and particle packing was assessed. Mineral contents were adequate for the sepiolites (> 90% of main mineral) and smectites (78 and 95%), but should be improved on the palygorskite samples (around 50% of main mineral in two of the studied samples). According to the textural characterization, the samples presented almost equal size distributions, but exhibited two different morphologies (i.e., laminar for smectites and fibrous for sepiolites and palygorskites). Both sepiolites possessed high flowability, whereas this factor varied from medium to low in the other materials. The compressibility of smectites was adequate, whereas that of palygorskites was high and that of the sepiolites was low. All of the samples studied would be useful as solid dosage form excipients (some samples would require improvement of their flow properties or treatment to reduce the amount of mineral impurities). PMID- 10669918 TI - Characteristics of pharmaceutical grade phyllosilicate compacts. AB - The purpose of this paper was to determine the possible employment of three groups of phyllosilicates (two smectites, three palygorskites, and two sepiolites) as direct-compression tablet excipients. Compact lots of each material were obtained after application of three different compression loads and the elastic recoveries and tensile strengths of each lot were then determined. The results showed that, in comparison with laminar minerals (smectite samples), fibrous minerals (palygorskites and sepiolites) were found to have lower elastic recovery, regardless of the compression load. Almost all elastic recovery happened immediately after the ejection of the compacts. Then, and at least during the next 24 hr, smectite compacts maintained their dimensions without any significant variation, whereas fibrous compacts tended to continue increasing slightly their dimensions. On the other hand, compacts made with laminar particles had higher mechanical strength than those made with fibrous particles, except for one sepiolite sample in which the strength was similar. Both particle morphologies and the presence or absence of hydration water molecules were taken into account in order to explain these differences. According to the compact properties measured, at least two of the noncommercial samples (smectite from Gador, Spain, and sepiolite from Vicalvaro, Spain) would be useful as direct compression tablet excipients. PMID- 10669919 TI - Tablet and capsule hydrophilic matrices based on heterodisperse polysaccharides having porosity-independent in vitro release profiles. AB - The purpose of the study was to investigate the release-controlling action of a swellable hydrophilic material based on heterodisperse polysaccharides (HP) in relation to the initial pore structure of the formulations. HP-based granules were produced under carefully controlled conditions and compacted into matrix tablets having equivalent tablet thickness. Quantification of pore structure using mercury porosimetry showed that the tablets had substantially different pore volumes and pore size distributions. Dissolution studies demonstrated that release of a water-soluble model compound, benzamide, from swollen matrices was affected neither by total porosity nor median pore diameter of the initial dry matrix. To extend the concept of porosity-independent release further, HP-based formulations containing either diclofenac sodium or propranolol HCl were contained within hard gelatin capsules in the form of uncompacted granules. This produced a dosage form with a high intraparticulate porosity in the dry state. Equivalent weights of the same formulations were also compacted into tablets. The in vitro release profiles from matrix tablets compacted from any of the formulations did not differ significantly from release profiles obtained when the same materials were contained uncompacted in hard gelatin capsules. PMID- 10669920 TI - Waxy corn starch: a potent cofiller in pellets produced by extrusion spheronization. AB - The purpose of this study was to assess the usefulness of waxy corn (maize) starch as a cofiller and diluent in pellets produced by aqueous extrusion spheronization. Waxy corn starch was combined with microcrystalline cellulose (MCC) in the range of 20-50% of the entire composition. Pellets containing ordinary corn starch or lactose with MCC were used as reference. The shape of pellets was characterized using an optical microscopic image analysis system. The surface and cross-sectional structure were investigated by means of scanning electron microscopy (SEM). The replacement of ordinary corn starch by waxy corn starch made it possible to increase the amount of starch from 20 to 40%. The pellets containing 50% waxy corn starch were of poorer quality but superior to those containing 30% corn starch. The surface structure became slightly more irregular with respect to the amount of either starch, and a cavity was formed inside the pellet during the spheronization. The origin of starch did not affect the surface structure of the pellets. Waxy corn starch is a potential cofiller: the amount of MCC can be reduced in pellets produced by extrusion-spheronization by using waxy corn starch as a cofiller. This enables the reduction of the manufacturing cost of pellets with low drug load. PMID- 10669922 TI - Color as an indicator of the organization and compactibility of binary powder mixes. AB - The aim of this study was to relate the color of several binary mixes to their organization as observed by scanning electronic microscopy, and to their compactibility. Binary mixes of niflumic acid (yellow) with ethyl cellulose, hydroxypropylmethylcellulose, low-substituted hydroxypropylcellulose (L-HPC), and ibuprofen (all white) were prepared using different particle size ranges. Colors of the mixes were determined by diffuse reflectance spectroscopy using a chromameter. Linear correlation was observed between the yellowness index/whiteness index ratio (Y/W ratio) defined by the American Society for Testing and Materials (ASTM) standards and the mean particle size difference of the materials which governs the organization of the blend. Except for the least interacting mix, the niflumic acid/L-HPC series, the color of the blend was also related to the tensile strength of the tablets made from the binary mixes. Color could be an interesting indicator of the organization of a powder mix. Diffuse reflectance spectroscopy could be used as a quality control tool because any modification of the color of the mix may be an indicator of a modification of its compactibility. PMID- 10669923 TI - Adsorption of esters of p-hydroxybenzoic acid by filter membranes: mechanism and effect of formulation and processing parameters. AB - The adsorption of esters of p-hydroxybenzoic acid (parabens) by filter membranes was studied by a flow-through technique. The hydrophobic effect was the major mechanism of adsorption. Propylparaben was more extensively adsorbed by all the membranes than was methylparaben. Hydrophobic membranes exhibited the greatest degree of adsorption, whereas adsorption was minimal for hydrophilic membranes. The charge of the filter membrane did not affect paraben adsorption. Formulation factors studied included the concentration of paraben, the presence of a tonicity modifying agent (sodium chloride, mannitol, glycerin), and the presence of a chelating agent (edetate sodium). Paraben adsorption was directly related to paraben concentration. The presence of a tonicity modifier or chelating agent did not alter paraben adsorption to membrane filters. Processing parameters studied included flow rate, temperature, autoclaving, flow interruption, and filter membrane presaturation. Presaturation of the filter membranes for up to 1 hr reduced but did not eliminate paraben adsorption during simulated use. Interrupting the flow of the paraben solution through the filter membrane allowed for additional paraben adsorption and caused the concentration of paraben in the restarted filtrate to be less than 100% of theory. PMID- 10669921 TI - Mucoadhesive DL-lactide/glycolide copolymer nanospheres coated with chitosan to improve oral delivery of elcatonin. AB - The purpose of this work was to develop a novel mucoadhesive DL-lactide/glycolide copolymer (PLGA) nanosphere system to improve peptide absorption and prolong the physiological activity following oral administration. The desired PLGA nanospheres with elcatonin were prepared by the emulsion solvent diffusion method to coat the surface of the resultant nanospheres with a mucoadhesive polymer such as chitosan, poly(acrylic acid), and sodium alginate. Their mucoadhesive properties were evaluated by measuring the nanospheres adsorbed to a rat everted intestinal sac (in vitro). The chitosan-coated nanospheres showed higher mucoadhesion to the everted intestinal tract in saline than the other polymer coated nanospheres. There was no mucoadhesion site-specificity of the chitosan coated nanospheres between duodenal, jejunal, and ileal sacs. The payload of drug in the chitosan-coated nanospheres was successfully increased by using the solvent diffusion method in oil. The pattern of drug release of the resultant nanospheres did not differ markedly from that of uncoated nanospheres. The chitosan-coated nanospheres with elcatonin were administered intragastrically to fasted Wistar rats. The chitosan-coated nanosphere reduced significantly the blood calcium level compared with elcatonin solution and uncoated nanospheres, and the reduced calcium level was sustained for a period of 48 hr. Even under nonfasting conditions, the mucoadhesion of chitosan-coated nanospheres was unaltered and the reduction in blood Ca levels was maintained satisfactorily. PMID- 10669924 TI - Hydraulic high-pressure nebulization of solutions and dispersions for respiratory drug delivery. AB - The purpose of this investigation was to assess hydraulic high-pressure nebulization as a means for respiratory drug delivery. A hydraulic high-pressure nebulizer was designed and constructed. In a design study, the output efficiency and the aerosol particle size were determined for the nebulizer as a function of nozzle diameter (5, 10, and 20 microns), gas flow rate (2 and 8 l/min), applied hydraulic pressure (2200 and 4000 psig), and distance between the nozzle orifice and impaction surface (0.25-4 cm) with an aqueous solution of fluorescein. The output efficiency was also measured with an ethanol solution and an aqueous phospholipid dispersion of liposomes. For the design study, each factor had an effect. The efficiency tended to increase with a decrease in the nozzle diameter, although the differences between the 5- and 10-micron nozzle were more sensitive to the air flow rate and nozzle-to-impaction-surface distance. Greater efficiencies were always obtained at the higher ancillary air flow rates. Operating the nebulizer at different pressures caused a change in the functional relationship between the efficiency and the nozzle-to-impaction-surface distance. For the 5-micron nozzle at high pressure, efficiency fell with increasing nozzle to-impaction-surface distance, whereas for the data obtained with the 20-micron nozzle, the efficiency increased with nozzle-to-impaction-surface distance, with lower efficiencies obtained at the higher pressures. For the remaining observations made with the 5- and 10-micron nozzles, the efficiency as a function of nozzle-to-impaction-surface distance appeared to be variable. For the 5- and 10-micron size nozzle, there was no significant effect of the air flow rate, pressure, or nozzle-to-impaction-surface distance on the mass median aerodynamic diameter and geometric standard deviation. For the 20-micron size nozzle, the particles were not completely dried. Ethanol solutions gave somewhat higher efficiencies, whereas the phospholipid dispersion gave efficiencies comparable to the aqueous solutions nebulized under similar conditions. The efficiency of the hydraulic high-pressure nebulizer appears to be correlated with the calculated properties of the liquid jet. For respiratory drug delivery, the hydraulic high pressure nebulizer provides reasonably high outputs of respirable particles independent of time from a single pass of liquid through the nebulizer. PMID- 10669925 TI - The effect of lyophilization on plasmid DNA activity. AB - The effect of lyophilization of plasmid DNA's ability to express an encoded protein was studied. Plasmid DNA, pRL-CMV expressing Renilla luciferase, was purified and stored in Tris-ethylenedi-aminetetraacetic acid (EDTA) buffer. Aliquots of the plasmid were lyophilized using analytical equipment, both alone and in the presence of carbohydrate. Samples were rehydrated and subject to functional and structural analyses. Analytical techniques included transfection efficiency in COS-1 cells, agarose gel electrophoresis, dimethylethylenediamine (DMED) assay for abasic sites, circular dichroism measurement, and UV spectroscopy. The lyophilization of pRL-CMV plasmid DNA resulted in a statistically significant loss of transfection efficiency (p < 0.05). Mono- and disaccharides could completely restore transfection efficiency. Agarose gel electrophoresis and the DMED assay demonstrated no change in gross plasmid structure or increase in abasic sites during lyophilization, respectively. Changes in DNA form, as measured by a change in ellipsisity, were observed on lyophilization. However, these changes were transient and were not shown to be responsible for loss of transfection efficiency. A hyperchromic effect was observed at 260 nm after lyophilization and could be reversed by the presence of carbohydrates. Lyophilization causes a decrease in plasmid DNA activity as measured by an in vitro transfection assay. Carbohydrates can ameliorate this decreased activity, which may be due to structural changes seen during the lyophilization process. PMID- 10669926 TI - Assessment of the myotoxicity of pharmaceutical buffers using an in vitro muscle model: effect of pH, capacity, tonicity, and buffer type. AB - The purpose of the present study was to investigate the myotoxicity of three buffers containing carboxylic acid groups (i.e., acetate, succinate, and citrate) as a function of their pH, capacity, and tonicity. The myotoxicity of these buffers in the range of pH 2-6 and 0.001-0.1 M buffer capacity was assessed using cumulative creatine kinase (CK) release from an isolated rodent muscle model following injection. Phenytoin and 0.9% NaCl injection were used as positive and negative controls, respectively. Buffer solutions were prepared. A lower pH and higher buffer capacity was linked to increased myotoxicity for the acetate buffers. However, for succinate and citrate buffers, pH appeared to influence the extent of myotoxicity, whereas buffer capacity did not seem to have an effect. When either NaCl or trehalose was used as a tonicity-adjusting agent at pH 6, isotonic 0.01 M buffer solutions dramatically lowered the cumulative CK release compared to those that were not isotonic. Isotonic succinate buffers displayed the lowest myotoxicity, whereas citrate buffers displayed the highest values. Citrate buffers containing three carboxylic acid groups showed higher myotoxicity than succinate buffers and acetate buffers at 0.001 and 0.01 M buffer capacities, whereas acetate buffer produced higher cumulative CK release than citrate and succinate buffers at 0.1 M buffer capacity. The myotoxicity of pharmaceutical buffers containing carboxylic acid groups appears to be directly affected by lowering the pH of the solution. PMID- 10669927 TI - Influence of drying temperature and granulation liquid viscosity on the inter- and intragranular drug migration in tray-dried granules and compacts. AB - The influence of the drying temperature and granulation liquid viscosity on the inter- and intragranular migration of a poorly water-soluble compound in a granulation mass and in a compact was quantitatively assessed. The intergranular migration kinetics were investigated by evaluating the drug distribution at different drying-time intervals. The results were analyzed by use of two-factor, three-level, face-centered, central composite designs. Riboflavin was mixed with alpha-lactose monohydrate 90 M and granulated with distilled water, except for the viscosity experiments in which an aqueous polyvinylpyrrolidone (PVP) (Kollidon K90) solution was used. The wet granules were dried in a hot-air oven or compacted prior to drying. The drug concentration at different locations inside the granulated mass and the compacts after drying was determined spectrophotometrically and by use of diffuse light reflectance measurements. The riboflavin distribution in the granulation masses and in the compacts was characterized by drug-enriched outer layers and drug-depleted inner regions, indicating a strong migration phenomenon. It was clear that the drying temperature had no influence on the inter- and intragranular drug distribution. The intergranular migration was avoided using the PVP as a binder in the granulation liquid, whereas a minimal granulation liquid viscosity of 100 mPa.sec was necessary to avoid the intragranular migration. The diffuse light reflectance measurements can be used for the in-process control of granule samples containing low drug concentrations without the destruction of the samples. PMID- 10669928 TI - Biochemical and morphological identification of ceramide-induced cell cycle arrest and apoptosis in cultured granulosa cells. AB - We have investigated the effects of ceramide on the progression of cell cycle and on apoptotic cell death in ovarian cultured granulosa cells. Rates of cellular proliferation were measured by immunocytochemical staining for proliferating cell nuclear antigen (PCNA) and flow cytometric cell cycle analysis. We also examined for morphological and biochemical signs of apoptosis. The PCNA expression was downregulated in a dose-dependent manner after treatment with C6-ceramide. Flow cytometric analysis demonstrated that the exposure of granulosa cells to C6 ceramide markedly decreased the population associated with G0/G1 DNA content and the reduction of cell numbers in G0/G1 phase was accompanied by the elevation of the A0 phase. The exposure of granulosa cells to exogenous C6-ceramide induced drastic morphological changes including cytoplasmic- or nuclear condensation and typical apoptotic DNA degradation. We also observed that phorbol 12-myristate 13 acetate, a protein kinase C (PKC) activator, significantly inhibited the ceramide induced apoptosis. These results suggested that ceramide might block the progression of cell cycle at G0/G1 phase and as a consequence, granulosa cells would be committed to apoptosis. Our findings also indicated that down-regulation of the PKC activity might be involved in the ceramide-induced apoptosis in cultured granulosa cells. PMID- 10669929 TI - Increased expression of VE-cadherin correlates temporally with differentiation of a restrictive endothelial barrier during normal angiogenesis in vivo. AB - The purpose of this study was to evaluate temporal expression of VE- and N cadherins within the angiogenic chick chorioallantoic membrane (CAM). Whether their relative patterns of expression changed in conjunction with abrupt differentiation of the restrictive CAM endothelial barrier between days 4.5 and 5.0 of the 21 days gestation was evaluated. Immunoblots against VE-cadherin depicted an increase of VE-cadherin expression between days 4.5 and 5.0, but no change in expression was detected between days 5.0 and 6.0. N-cadherin expression, on the other hand, remained uniform from day 4.5 to day 6.0. Immunogold-labeled anti-VE-cadherin was found exclusively on the CAM endothelium, and principally along the lateral inter-endothelial junctions. Hence, VE-cadherin expression by the angiogenic endothelium was similar to that of adult endothelium. That VE-cadherin expression by the CAM endothelium was increased between days 4.5 and 5.0 serves to suggest a temporal correlation with the ontogeny of restrictive barrier function in angiogenic endothelium in vivo. PMID- 10669930 TI - Human chondrocyte cell lines from articular cartilage of metatarsal phalangeal joints. AB - Chondrocytes can be isolated from human adult cartilage from metatarsal phalangeal joints. After enzymatic digestion to isolate viable cells, confluent monolayers were obtained 2-4 weeks after the start of cell division. Chondrocytes cultures, initiated and maintained in HAM's F12 with bovine fetal serum without the addition of other growth factors, produced in vitro a matrix rich in collagen and proteoglycans. Although several studies reported phenotypic instability, our results showed that the cell retain for more than 5 months in culture their differentiated characteristics, including the ability to produce cartilage specific molecules. Chondrocyte cell lines should be useful in studying the functions of these cells from normal and abnormal tissue and for pharmacological studies in vitro. PMID- 10669931 TI - Tight junction of sinus endothelial cells of the rat spleen. AB - The fine structure of the tight junctions between sinus endothelial cells of the rat spleen and the permeability of such sinus endothelial cells were examined by transmission electron microscopy, using freeze-fracture, triton extraction, and lanthanum-tracer techniques. In freeze-fracture replicas, the segmented strands and grooves of the tight junctions were frequently observed on the basolateral surfaces of the sinus endothelial cells irrespective of the location of the ring fiber. There were one or two wavy-strands or grooves which were, for the most part, oriented parallel to the long cell axis thus forming networks at places. In addition, some strands or grooves were discontinuous while some networks of the junctional strands were not closed. These strands also occasionally lacked intramembranous particles in the tight junctions. The junctional strands run apicobasically at certain sites. In the vertical sections of the sinus endothelial cells treated with lanthanum nitrate, although no tight junctions were observed wherever the endothelial cells were apposed, most of them were situated on the basal part of the lateral surfaces of the adjacent endothelial cells. Several fusions of the junctional membranes were observed in a vertical section of the lateral surfaces of the adjacent endothelial cells. The intercellular spaces of the adjacent endothelial cells except for the fusion of the junctional membranes, were electron dense and the infiltration of lanthanum nitrate was found not to be interrupted by these tight junctions. Based on these observations, the molecular 'fence' and paracellular 'gate' functions of the tight junctions in the sinus endothelial cells are discussed. PMID- 10669932 TI - Effects of prolonged ACTH-stimulation on adrenocortical accumulation of lipofuscin granules in aged rats. AB - Subcellular deposition of lipofuscin granules is a marker of aging. Human and rodent adrenal cortices accumulate lipofuscin granules with age, but the mechanism that leads to the accumulation is not known. The ultrastructural appearance of lipofuscin granules resembles that of secondary lysosomes. Since adrenocortical subcellular events are predominantly influenced by ACTH action, we therefore studied the effect of prolonged ACTH-stimulation on adrenocortical accumulation of secondary lysosome-like granules, designated herein as lipofuscin granules. Using aged Fischer 344 male rats as a model, we found that a 7 day ACTH stimulation exerts a reducing effect on adrenocortical lipofuscin accumulation. Thus, adrenocortical accumulation of lipofuscin granules with age in vivo may not be an irreversible process. PMID- 10669933 TI - Desmosomal proteins in cultured and intact human periodontal ligament fibroblasts. AB - This study examined the kinds of desmosomal proteins in the human periodontal ligament fibroblasts (PDLFs). The PDLFs obtained from young and older patients were cultured and the amounts of desmosomal proteins were measured by ELISA with antibodies to desmoplakins, desmogleins, and desmocollins. Cultured cells and tissue sections of the human periodontal ligament were immunostained with the same antibodies. Expression of desmosomal proteins in the PDLFs was clearly demonstrated both by ELISA and the immunohistochemical studies, suggesting the existence of desmosome-like junctions in the PDLFs. The junctions are considered to protect gap junctions in the PDLFs against cell transformation caused by cell contraction, which may relate to tooth eruption and repair of periodontal tissue, and/or strong occlusal forces. Statistically significant differences (P < 0.0001) in the expression of desmoplakins and desmogleins between younger and older patients were observed in this study. PMID- 10669934 TI - Distribution of actin bundles in Bowman's capsule of rat kidney. AB - In this study we define the distribution of actin bundle arrangement in Bowman's capsule of rat renal corpuscles. Parietal cells of Bowman's capsule were examined by conventional light microscopy, electron microscopy and confocal microscopy. Within each parietal cell individual actin bundles are arranged in a parallel fashion running the length of the cell. Computer reconstructions obtained using confocal microscopy clearly show the lengths of actin bundles to be arranged, on a capsule level, end-to-end, at angles and perpendicular to bundles in adjacent cells. The bundles stain positively for non-muscle myosin and vinculin. The presence and arrangement of actin bundles in parietal cells is consistent with a role in reinforcing capsule structure. PMID- 10669935 TI - Immunolocalization of androgen receptors in testicular cells during postnatal development of the bank vole (Clethrionomys glareolus, S.). AB - Determination of the cellular distribution of the androgen receptors within the testis is of great importance for an understanding of their essential role in mediating of androgen action in the male gonad. In bank voles, which are seasonally breeding rodents, photoperiod is one of the most important factors inducing profound changes in the morphology and hormonal activity of the testes. Immunolocalization of androgen receptors was found in all somatic cells such as Sertoli cells, Leydig cells, and peritubular-myoid cells, however, distribution of the androgen receptors in various cell types depended on age of animals. Intensity of immunoreactivity was noticed as age and photoperiod-dependent. Males reared under different light regimes showed a significant correlation between the length of light and sexual maturation. Therefore, morphology of the testis from young and adult bank voles was also presented. PMID- 10669936 TI - The current prospects for neutrophil transfusions for the treatment of granulocytopenic infected patients. AB - Infection continues to be a major cause of morbidity and mortality in patients undergoing aggressive chemotherapy and hematopoietic stem cell transplantation. The provision of normal neutrophils to such patients is a logical therapeutic approach, the success of which will likely be dependent on the dose of neutrophils provided. Stimulation of normal leukapheresis donors with G-CSF or dexamethasone causes marked neutrophilia and results in the collection of greatly increased numbers of neutrophils. Transfusion recipients, on average, exhibit large posttransfusion neutrophil increments that are sustained for 24 hours. These cells are capable of migrating to extravascular sites. Although the preliminary clinical impressions are encouraging, the clinical efficacy of transfusing large numbers of neutrophils will have to be determined by randomized controlled clinical trials. PMID- 10669937 TI - Mechanisms and new approaches for the allogeneic blood transfusion-induced immunomodulatory effects. PMID- 10669938 TI - Prestorage versus poststorage white cell reduction for the prevention of the deleterious immunomodulatory effects of allogeneic blood transfusion. PMID- 10669939 TI - Leukocyte reduction of blood components: public policy and new technology. PMID- 10669940 TI - Camouflaged blood cells: low-technology bioengineering for transfusion medicine? AB - The small number of studies done on the covalent modification of RBC with PEG, or PEG-derivatives, suggests that the immunocamouflage of intact cells significantly reduces the antigenicity and immunogenicity of the foreign cell. Importantly, this protective immunologic effect can be accomplished without adversely affecting the structure, function, or viability of the modified cell (e.g., RBCs and lymphocytes). As a consequence, PEG-RBC may have significant practical value in the treatment of the chronically transfused patient as a prophylactic measure against allosensitization. The PEG-RBC also may be useful in treating the already allosensitized individual. As shown, preexisting antibodies do not effectively recognize nor bind to the modified donor cells. A finding of further interest to transfusion medicine is that pegylation of contaminating lymphocytes within RBC products may prove efficacious in preventing graft-versus-host disease in the immunocompromised patient. However, the main emphasis of our research continues to be the immunocamouflage of RBC for use in chronic transfusion therapy of the SCD and thalassemic patient. PMID- 10669941 TI - Risk of hepatitis A virus infection in persons with hemophilia receiving plasma derived products. PMID- 10669942 TI - What we can learn from national and international platelet serology workshops. PMID- 10669943 TI - Position-independent expression of transgenes in zebrafish. AB - The variability in expression patterns of transgenes, caused by the influence of neighboring chromatin, is called 'position effect'. Border elements are DNA sequences, which have the ability to alleviate position effects. The abilities of two types of border elements, scs/scs' from the D. melanogaster 87A7 heat shock locus and the A-element from the chicken lysozyme gene, to protect transgenes from position effects were quantified in developing zebrafish embryos. The transgenic construct used was FV3CAT, which consists of the carp beta-actin transcriptional regulatory region, the chloramphenicol acetyltransferase (CAT) gene and the 3'-untranslated region from the Chinook salmon growth hormone gene. FV3CAT constructs flanked by either scs/scs'-elements or A-elements were introduced into zebrafish chromosomes and the spatial and temporal expression patterns of the transgenes were quantified in multiple generations of transgenic zebrafish. Levels of transgene expression were uniform in the pre-differentiated and fully differentiated populations of cells present during embryonic development. Levels of transgene expression were proportional to the numbers of integrated transgenes. Expression of transgenes per cell varied less than two fold in different transgenic lines. Both types of border elements were able to prevent the influences of neighboring chromatin on transgene expression through three generations of fish. The results are consistent with the ability of border elements to function with equal efficiencies in the many cell types found in vertebrates. Thus, inclusion of border elements in genetic constructs can provide reliable and reproducible levels of gene expression in multiple lines of fish. PMID- 10669944 TI - Zona pellucida glycoprotein mZP3 produced in milk of transgenic mice is active as a sperm receptor, but can be lethal to newborns. AB - Mouse egg zona pellucida glycoprotein mZP3 (approximately 83 kDa M(r)) serves as a species-specific sperm receptor and acrosome reaction-inducer during fertilization in mice. These biological activities are dependent on certain mZP3 serine/threonine- (O-) linked oligosaccharides present at the combining-site for sperm. In an attempt to produce large amounts of biologically active mZP3, we generated several transgenic mouse lines carrying the full-length mZP3 gene fused to the beta-casein gene promoter and transcription termination sequence. We found that different transgenic mouse lines have different amounts of recombinant mZP3 (approximately 63 kDa M(r)) in milk of lactating females, from approximately 0.3 to 3.5 micrograms/microliter of milk. In all cases, purified milk-mZP3 is active as a sperm receptor and acrosome reaction-inducer in vitro. Unexpectedly, we also found that development of litters from these transgenic mice is related to the amount of mZP3 in the mother's milk. In the most extreme case, litters from the highest expressers fail to live beyond about day-7 post partum unless placed immediately after birth with surrogate wild-type mothers. Litters from lower expressers initially display a complex phenotype that includes effects on hair and body growth, but some of the mice survive and, in time, are restored to a wild-type phenotype. These results demonstrate that relatively large amounts of biologically active mZP3 can be produced in transgenic mouse milk for structural and other studies, but that the presence of mZP3 in milk has dramatic developmental effects on litters, ranging from retarded hair and body growth to death of newborn pups. PMID- 10669945 TI - Codon optimization, genetic insulation, and an rtTA reporter improve performance of the tetracycline switch. AB - The objective of this work was to further develop a tetracycline repressor (TetR) protein system that allows control of transgene expression. First, to circumvent the need for a binary approach, a single plasmid design was constructed and tested in tissue culture. To indirectly assay integrations that express the synthetic transcription factor (rtTA), a bicistronic gene was built which included an internal ribosome entry site (IRES) and a green fluorescent protein coding region (GFP) on the same expression cassette as the coding region of rtTA (pTetGREEN). This construct did not produce fluorescent colonies when stably integrated and provided minimal expression of GFP in the face of adequate expression of rtTA. The coding region for TetR was then altered by introducing 156 silent point mutations to simulate mammalian genes. Replacement of wild-type TetR gene (tetR) in pTetGREEN with 'mammalianized' tetR provided GFP expression. Adjustment of codon usage in the tetR region of rtTA nearly doubled the expression level of functional rtTA. To increase the number of rtTA expressing lines, the chicken egg-white lysozyme matrix attachment region (MAR) was introduced into the single plasmid design just upstream of the tetracycline operators (tetO). Inclusion of the MAR doubled the number of colonies that expressed rtTA (44% vs 88%). With the modifications described here, the number of lines that express rtTA and provide induction from a single plasmid design can be increased by the inclusion of a MAR and the level of rtTA expression can be further increased by adjusting the base composition of the TetR coding region. The MAR also insulates the inducible gene from the promoter driving rtTA. PMID- 10669946 TI - Production of transgenic rats using cryopreserved pronuclear-stage zygotes. AB - We investigated the application of cryopreserved pronuclear-stage zygotes for the production of transgenic rats. Most of the pronuclear-stage zygotes cryopreserved by conventional two-step freezing or vitrification appeared morphologically normal, but the proportion of frozen zygotes that developed into fetuses following transfer (59.7-60.2%) was higher than that of vitrified zygotes (5.5 22.1%). When the frozen-thawed zygotes were used for DNA microinjection, 97.5% survived after DNA microinjection and 25.1% of the transferred zygotes developed into fetuses. These proportions were comparable to those of the fresh control zygotes (97.0 and 30.0%, respectively). The integration efficiency of the exogenous DNA into fetuses was similar between the frozen group (3.3% per injected zygote) and the control group (3.5%). These results indicate that pronuclear-stage rat zygotes can be successfully cryopreserved by conventional two-step freezing for production of transgenic rats. PMID- 10669947 TI - Angiogenesis at the interface between basic and clinical research. AB - The field of antiangiogenesis has shown a remarkably rapid evolution from the discovery at the bench to translation into the clinic. Currently a wide variety of compounds are in clinical trial as inhibitors of angiogenesis, and new compounds are being frequently added. The target cell of most angiogenesis inhibitors is the endothelial cell, with inhibitors that selectively affect a number of endothelial cell functions acquired during angiogenesis, including activation, proliferation, migration, invasion and survival. The endothelial cell may also be targeted by chemotherapeutic agents currently in use. The high doses and intermittent treatment schedules used to fight resistant tumor cells may be altered towards lower doses and chronic administration to obtain selective inhibition of angiogenic factor-stimulated endothelial cells as adjuvant therapy. Finally, gene therapy is a promising route for the delivery of novel protein inhibitors of angiogenesis, and is actively being investigated. PMID- 10669948 TI - Models for studying angiogenesis in vivo. AB - In vivo and in vitro techniques are available for research on the functions of endothelial cells during angiogenesis. In this review we describe and evaluate the methodology and specific features of some of the most frequently used in vivo assays. PMID- 10669949 TI - Biological parameters for the choice of antiangiogenic therapy and efficacy monitoring. AB - Angiogenesis is a tightly controlled process which depends on the balance between stimulating and inhibiting factors. When this balance is disrupted, angiogenesis acquires a pathological meaning. The list of molecules able to induce angiogenesis is heterogeneous with respect to their chemical characteristics and biological properties. Quantitative measurement of tumor angiogenesis is necessary for the choice of therapeutic strategies and as an endpoint for antiangiogenic therapy. We are developing a quantitative RT-PCR with measures the expression of specific factors in real time. With the use of this rapid technique, measurement of the expression of the angiogenic factors and inhibitors is also possible in specimens as small as biopsies. PMID- 10669950 TI - Assessment of tumor vascularization: immunohistochemical and non-invasive methods. AB - Growth of solid tumors beyond a certain mass is dependent on the vascular bed from pre-existing host vasculature. The process of angiogenesis is essential not only for primary tumor growth but also for metastasis. The number of microvessels within the invasive component of a primary tumor reflects the degree of tumor angiogenesis. At present the most widely used method to assess neovascularization is the quantitation of intratumoral microvessel density (IMD) by immunohistochemical methods in which specific markers for endothelial cells are employed. In this paper we analyze the different methods used to assess IMD, as well as their advantages and potential methodological pitfalls. Several studies have shown a close correlation between IMD, tumor growth and the occurrence of metastasis, suggesting that IMD is a prognostic indicator of clinical relevance. Furthermore, preliminary studies suggest that determination of angiogenesis may predict responsiveness to some forms of conventional anticancer therapy. Although the histological microvessel density technique is the current gold standard to characterize tumor angiogenesis, it may not be the ideal tool for clinical purposes because it needs to be performed on biopsy material and does not assess the functional pathways involved in the angiogenic activity of tumors. Non invasive assessment of tumor vascularity is possible in vivo by means of Doppler sonography, dynamic contrast-enhanced magnetic resonance imaging (MRI) and positron emission tomography (PET). These methods may be preferable to histological assay because they are non-invasive, survey the entire tumor, reflect both anatomic and physiologic characteristics, and may be useful to monitor the activity of antiangiogenic therapies. PMID- 10669951 TI - MMP inhibitors: experimental and clinical studies. AB - Matrix metalloproteases (MMPs) are a family of structurally related enzymes that are capable of degrading proteins of the extracellular matrix. These enzymes play a role in tissue remodelling associated with both physiological and pathogenic processes. A high expression of MMPs is associated with cancer malignancy: it is related to the tumor's ability to metastasize and to the process of angiogenesis. Treatment with MMP inhibitors alone or in combination with cytotoxic therapy is an interesting novel approach to control tumor progression. The expected mechanism of action of these compounds and the difference in side effects compared to cytotoxic drugs make the definition of endpoints and the assessment of response difficult. Furthermore, it is not yet clear whether tumor vascularization or, more specifically, MMP expression/activation should be a criterion of eligibility for this kind of treatment. This review provides an overview of the characteristics of MMPs and their role in tumor progression, metastasis and angiogenesis. Preclinical and clinical studies with synthetic MMP inhibitors are described. The presence of MMPs in biological fluids of patients and their use in prognostic evaluation and in determining the efficacy of treatment with MMP inhibitors is discussed. PMID- 10669952 TI - Angiosuppression and chemotherapy: strategies aimed at their integration in cancer patients. AB - A number of antiangiogenic agents have been developed as pharmaceuticals and are currently being tested in clinical studies. Potential strategies to enhance the activity of angiogenesis inhibitors could be to combine them, or better still, to administer them either sequentially or concurrently with cytotoxic drugs. Chemotherapy would be a more appropriate initial choice for patients with advanced disease since cytostatic agents can induce a fast regression of the tumor and cancer-related symptoms. Antiangiogenic treatment could be used after chemotherapy in patients who achieve disease remission to prolong the time to progression, the symptom-free interval and the overall survival. Antiangiogenic treatment is likely to attain an important role in the adjuvant setting. In fact, it could be used for prolonged periods after radical surgery to maintain dormancy of residual tumor cells. In spite of these promising preclinical data, several points need to be clarified before the initiation of clinical trials. In fact, certain misconceptions may interfere with their optimum design and result analysis. PMID- 10669953 TI - In vitro models of angiogenesis: the use of Matrigel. AB - Tumor-induced angiogenesis is a key event for neoplastic progression. In vitro assays are important for identification of potential angiogenic agents and rapid screening for pharmacological inhibitors. The increased interest in this field of study has generated several in vitro assays that recapitulate the steps of endothelial cell activation and differentiation. In this short report we emphasize the utility of Matrigel, a reconstituted basement membrane, to define two different steps in the angiogenic process: invasion in response to growth factors and organization of microvessels into a network with branching morphology on a Matrigel substrate. PMID- 10669954 TI - Quantitative RT-PCR assay for VEGF mRNA in human tumors of the kidney. AB - Angiogenesis is the formation of new capillaries from pre-existing vessels, and recent evidence has demonstrated that tumor growth is controlled mainly by angiogenesis. Vascular endothelial growth factor (VEGF) is an endothelium specific growth factor which is strongly angiogenic in vitro and in vivo. We have developed a quantitative RT-PCR assay for the measurement of VEGF mRNA expression using a real-time procedure based on the use of fluorogenic probes and the ABI PRISM 7700 Sequence Detector System. The assay performance of this method in terms of practicability and reliability is reported with results that seem promising for its widespread use in the clinical laboratory. The method has been applied to the measurement of mRNA of VEGF in human renal cell carcinomas (RCC). Preliminary results show a significantly higher VEGF mRNA expression (ratio values between 181 and 2222) in tumor specimens compared to non-adjacent, non tumoral tissue of the same subjects. PMID- 10669956 TI - Inhibition of angiogenesis by type I interferons in models of Kaposi's sarcoma. AB - Kaposi's Sarcoma (KS) is a pathology which occurs with increased frequency and in a particularly aggressive form in AIDS patients. The HIV-1 Tat protein appears to be an important co-factor in the induction of the extensive neo-vascularization associated with AIDS-KS. Tat acts as a chemoattractant for endothelial cells in vitro, inducing both chemotactic and invasive responses. Several clinical trials have been performed testing the effectiveness of diverse biological agents in therapy of KS, among these the type I interferons. Type I IFNs have diverse biological functions besides their anti-viral activity, including anti-angiogenic properties. We have shown that IFN alpha and IFN beta are potent inhibitors of both primary and immortalized endothelial cell migration and morphogenesis in vitro as well as neo-angiogenesis induced by HIV-1 Tat in vivo. The inhibitory effect of IFN class I on HIV-Tat associated angiogenesis further supports its use as a therapy for epidemic Kaposi's sarcoma. The use of recombinant IFNs at the levels required to obtain a therapeutic effect are associated with side effects and toxicity, therefore we are now developing a gene therapy approach for constant and local delivery type I IFNs. PMID- 10669955 TI - Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy. AB - Antiangiogenic therapy may represent a promising approach to cancer treatment. Indeed, the efficacy of endogenous angiogenesis inhibitors, including angiostatin, endostatin and TIMPs, has been demonstrated in many types of solid tumors in animal models. In view of the possible problems associated with long term administration of inhibitors as recombinant proteins, we propose their delivery as nucleic acids through a gene therapy approach. To this end, eukaryotic expression constructs for murine angiostatin and endostatin as well as human TIMP-2 were generated, and characterized in vitro. All constructs carry the relevant cDNAs under the control of the strong HCMV promoter/enhancer, and cleavable leader signals to allow protein secretion. Expression of the angiogenesis inhibitors was detected by in vitro transcription/translation experiments as well as transfection of 293T cells, followed by Western blotting (WB) or radioimmunoprecipitation analysis of both cell lysates and supernatants (SNs). These constructs might be used for in vivo intramuscular delivery of plasmid DNA and as a set of reagents for the development of retroviral as well as adeno-associated viral (AAV) vectors expressing angiogenesis inhibitors. PMID- 10669957 TI - Endostatin: a promising drug for antiangiogenic therapy. AB - Angiogenesis, the formation of new blood vessels from existing capillaries, is critical for tumors to grow beyond a few in size. Tumor cells produce one or more angiogenic factors including fibroblast growth factor and vascular endothelial growth factor. Surprisingly, antiangiogenic factors or angiogenesis inhibitors have been isolated from tumors. Some angiogenesis inhibitors, such as angiostatin, are associated with tumors while others, such as platelet-factor 4 and interferon-alpha are not. Endostatin, a C-terminal product of collagen XVIII, is a specific inhibitor of endothelial cell proliferation, migration and angiogenesis. The mechanism by which endostatin inhibits endothelial cell proliferation and migration is unknown. Endostatin was originally expressed in a prokaryotic system and, late, in a yeast system, thanks to which it is possible to obtain a sufficient quantity of the protein in a soluble and refolded form to be used in preclinical and clinical trials. PMID- 10669958 TI - The role of the thiol N-acetylcysteine in the prevention of tumor invasion and angiogenesis. AB - We have extensively studied the effects of N-acetylcysteine (NAC), a cytoprotective drug that can prevent in vivo carcinogenesis. Here we review our findings NAC completely inhibits gelatinolytic activity of metalloproteases and chemotactic and invasive activities of tumor cells. In addition, NAC reduces the number of lung metastases when malignant murine melanoma cells are injected into nude mice. NAC treatment decreases the weight of primary tumors and produces a dose-related increase in tumor latency. Moreover, oral administration of NAC reduces the formation of spontaneous metastases. In experimental metastasis assays, we have found a synergistic reduction in the number of lung metastases after treatment with doxorubicin (DOX) and NAC in nude mice. In tumorigenicity and spontaneous metastasis assays, the combined administration of DOX and oral NAC again has shown synergistic effects on the frequency and weight of primary tumors and local recurrences and completely prevented the formation of lung metastases. The addition of NAC to endothelial cells strongly reduces their invasive activity in response to angiogenic stimuli. NAC inhibited the degradation and release of radiolabeled type IV collagen by activated endothelial cells, indicating that NAC blocks gelatinase activity. Oral administration of NAC reduces the angiogenic response induced by KS tumor cell products, confirming the ability of NAC to inhibit the invasive activity of endothelial cells in vivo and thereby blocking angiogenesis. PMID- 10669959 TI - Student report--the fight against cancer and angiogenesis inhibitors: are we entering a new era? AB - Chemoresistance is currently the main cause of failure in the treatment of cancer which, despite extensive research, remains unsolved. In this report the theoretical assumptions underlying antiangiogenic therapy are described and future perspectives and limits are discussed. PMID- 10669960 TI - Psychoanalysis, psychoanalytic psychotherapy and supportive psychotherapy: contemporary controversies. AB - The author explores the controversies involving psychoanalytic psychotherapy from conceptual, clinical and educational perspectives. He proposes an integrated concept of psychoanalytic modalities of treatment, and their subdivision into standard psychoanalysis, psychoanalytic psychotherapy and psychoanalytically based supportive psychotherapy. Indications and contra-indications for these therapeutic approaches are outlined in the light of clinical experience and psychoanalytic research on these issues. It is proposed that psychoanalytic institutes teach psychoanalytic psychotherapy to candidates in psychoanalytic training. The author stresses that we now possess a broad spectrum of psychoanalytically based approaches to patients that significantly expand the therapeutic effectiveness of our profession, and thus can strengthen the social impact of psychoanalysis. PMID- 10669961 TI - Disappointment and disappointedness. AB - The author argues that the widespread affective experience of disappointment has not received the analytic attention it deserves, and that this is particularly the case for disappointedness as an outstanding feature of a way of life. Disappointedness is presented as a pathological organisation or character disorder that expresses specific unconscious fantasies and gives rise to disruptive transference-countertransference manifestations. The author singles out disappointedness as a special problem rather than, in the usual way, subsuming it under depressiveness or masochism and then assigning it a subsidiary or merely descriptive role. With the help of a case example, he attempts to illustrate the benefits of heightened clinical awareness of disappointment and disappointedness. These benefits include increased access to the compromise formations that can stand in the way of effective analytic work. PMID- 10669962 TI - Male gender identity and sexual behaviour. AB - One consequence of a heightened interest in intersubjectivity in the current psychoanalytic literature has been a relative neglect of the examination of unconscious fantasies. Presenting material from the analysis of three males, each of whom, in childhood and/or adolescence, hid his penis between his legs and looked at himself in a mirror naked, the author demonstrates the importance of attending to both unconscious fantasies and their manifestations within the interactive field of analysis. The first patient is a young child with a gender identity disorder, whose wish to be like his mother was a response to the emotional loss of her during early childhood. The second patient is an adolescent, whose behaviour in front of a mirror was a manifestation of his desire to possess his mother and be her, to humiliate and sadistically control her, and at the same time, to experience the masochistic sexual gratification of being a seemingly helpless victim. The third patient, a 48-year-old male, came to analysis filled with suicidal impulses and self-hatred related to homosexual impulses. His repeated examination of himself in a mirror, with penis hidden, reflected severe castration anxiety, related to an ambivalent relationship with an angry mother and a longing for attention from an unavailable father. The article closes with a description of the similarities and differences in the dynamics of these three males as well as a discussion of the meaning of similar behaviour in other males seen in consultation. PMID- 10669963 TI - Erotic complications. AB - The author argues that erotic transference-countertransference dynamics present particular complexities when they develop between gender constellations other than male analyst and female patient. She addresses the dynamics of a complicated erotic transference in concert with an aversive countertransference response as it evolved between a female analyst and female patient. The intense erotic transference that developed defied classification as either maternallerotic or oedipallerotic, and instead included both features in a rapidly shifting process that was difficult to address analytically. The analyst's confused, often aversive, response to her patient's erotic wishes ultimately revealed a subtle re enactment involving split-off and erotised experiences of emotional penetration and scrutiny. When these issues were addressed, the erotic transference dissolved, and the analyst's experience of her patient shifted rather dramatically. It is suggested that complex erotic transference sometimes contains within it evidence of previously repressed object experiences that were not primarily sexual in nature. PMID- 10669964 TI - Narcissistic resistances in the analytic experience. AB - The narcissistic resistances in the clinical experience and the supposed autonomy of the patient to reproduce the narcissistic stage are studied in this paper. The problem is examined through the analysis of a patient, who masturbated compulsively. The relationship between the symptom and the transference is investigated because the symptom related to specific behaviour from the object, and in order to understand the meaning of the phantasy that underlines the auto erotic activity. It is pointed out that, in order for defensive activity to fulfil its sense of narcissistic refuge, an other (distinct from the subject) must carry out an essential function of support of the narcissistic defence. Narcissistic self-sufficiency is governed by a paradox, as it needs an object to demonstrate that it can subsequently dispense with this object. A second paradox occurs when the narcissistic subject can only succeed in dispensing with the object if the object gives some sign indicating that he is affected by such 'doing without'. In the analytical relationship, this type of defence manifests itself as narcissistic resistance. The analyst participates in the covering-up of the dependency that the defensive system has on the external object and this is a prerequisite for establishing the resistance. PMID- 10669965 TI - The impact of a war experience on the inner world of a young child. AB - The author uses material from the treatment of a young child whose development became seriously arrested from the time of a traumatic experience during a war. This experience, combined with the impact of the birth of a younger sibling while he was still a baby himself, had catastrophic consequences. He could only communicate his feelings of terror and fear of dying through the use of projective identification. The persecutory anxiety of this child was such that for a while he could not tolerate hearing the interpretations, and was prone to violent outbursts. The author describes how she had to bear the projections and manage the physical attacks while trying to maintain the capacity to observe and think clearly. This setting produced an experience of containment that gradually allowed this child to accept and understand interpretations which diminished the power of feelings that had overwhelmed him in the past. Through introjection of the experience of being understood during his sessions, he began to develop the capacity to think about his feelings. This allowed him to gradually recover, develop, and make use of his intelligence and imagination, and the behavioural difficulties that brought him to treatment disappeared. PMID- 10669966 TI - 'My bad diagnostic error': once more about Freud and Emmy v. N. (Fanny Moser). AB - The author examines two aspects of the case of Fanny Moser (Emmy von N. in the 'Studies on Hysteria'): first, a hitherto unknown letter written by Sigmund Freud to Fanny Moser's daughter in 1935, in which he revises his diagnosis of half a century before, describing it as 'a bad diagnostic error' and apologising to the daughter; second, the dates of Fanny Moser's treatment. All previous attempts have failed to date it consistently. The author combines all the information about the case of Fanny Moser and the circumstances of Freud's life in that period and shows that Freud took over the case in 1888, not in 1889. In addition, he suggests that Emmy von N. has had a significant influence on Freud's papers on 'Hysteria' and on his comparative study 'Organic and hysterical motor paralyses'. PMID- 10669967 TI - Some reflections on identification. AB - The author presents a view of identification based on a rereading of two of Freud's key texts and an approach derived from an academic interpretation of Hegel dating from the 1930s. These aspects are considered at length. The importance of the human and anthropogenic element is stressed. The human subject is presented as coming into being through language; being called upon to be what he is not and not to be what he is, the subject appears as wishful in nature, desiring the wish of the other at the same time as he desires the object of the other's wish. The author argues that identification as a problem arises only in a human being who speaks or has received an injunction to speak; this raises the question of who or what he is and of being as such. Analytic treatment may in his view therefore proceed in one of two directions, one based on the interplay of projection and introjection with identification as an end, and the other on resistance and repression where the Oedipus complex is seen as the nuclear issue. Identification is seen in terms of overcoming the negative identity of not being all other subjects, and identity is found to be a conscious response that might even have a political element. PMID- 10669968 TI - Tender love and transference: unpublished letters of C. G. Jung and Sabina Spielrein. AB - The author dissents from the widely accepted interpretation that the relationship between Sabina Spielrein and Carl Jung in the years 1904-1910 included sexual intercourse and constituted an ethical breach of the doctor-patient boundary in the course of a treatment. Spielrein declared that her treatment ended with her discharge from the Burgholzli hospital as Jung's patient in 1904-1905. Jung maintained he 'prolonged the relationship' in order to prevent a relapse and also referred to it as a friendship. Materials published in 1994 (letters, drafts, diaries, hospital chart) and unpublished letters recently found by the author in the Claparede archive in Geneva shed new light on previously published documents and interpretations by Carotenuto that have dominated the secondary literature since 1980. The new materials provide a more nuanced view of the Spielrein-Jung relationship and point to the function of non-erotic love in the therapeutic relationship. A new look at the Freud-Jung correspondence about the Spielrein Jung relationship shows that Jung's perception that a sex scandal was initiated by Spielrein resulted from Jung's misreading of rumors concerning another woman; the episode had no ill effect on the relationship between Freud and Jung. PMID- 10669969 TI - Psychoanalysis and the neurosciences: a topical debate on dreams. AB - The author begins by pointing out that, whereas Freud first turned his attention to dreams in 1895, they became an object of neuroscientific interest only in the 1950s, after the discovery of rapid-eye-movement (REM) sleep and the observation that a subject woken in an REM phase could remember and narrate them. He discusses the various brain structures found by the neuroscientists to be implicated in dreaming and the associated hypotheses about their involvement in the processes of remembering dreams, their spatial construction and semantic organisation, and the dreamer's emotional participation in and narration of dreams. Attention is drawn to recent psychophysiological research findings indicating that dreaming occurs in all sleep phases and not only in REM episodes. The cognitivist contribution is also discussed. The author goes on to demonstrate the difference between the neuroscientific and psychoanalytic approaches to dreams. Whereas the neuroscientists are interested in the structures involved in dream production and in dream organisation and narratability, psychoanalysis concentrates on the meaning of dreams and on placing them in the context of the analytic relationship in accordance with the affective history of the dreamer and the transference. The brain structures and functions of interest to the neurosciences, while constituting the physical and biological substrate of these aspects, are stated to be irrelevant to their psychoanalytic understanding. PMID- 10669970 TI - The importance of capacities in psychoanalysis and the language of human development. AB - The author explores the human capacity to contain or hold experience--for self and other--by analysing the word capacity itself. Underlying the discussion is the proposition that hidden in the word capacity is a particular perspective on mental mechanisms fundamental to object-relations theory and that specific consideration of the word may, therefore, add to an understanding of these mechanisms. The author suggests that the term may have entered the language of psychoanalysis 'by mistake'. He then looks at its meaning in terms of its etymology, grammatical 'flavour', metaphorical significance and conceptual use, in particular by Bion and Winnicott. It is argued that psychoanalysis is part of, and makes a significant contribution to, a long tradition of learning from experience, in which development is understood as the progressive emergence or evolution of capacities. The underlying meaning of the word suggests that development is expansive rather than linear, step-by-step or cyclical (unlike, for example, stage, level, phase, position or loop). The author suggests that the notion of capacities may provide a framework for applying insights from the theory and practice of psychoanalysis in other organisational and societal contexts. PMID- 10669971 TI - Knowledge, consensus and uncertainty. AB - Some months ago the editors of this journal asked me if I would undertake a series of short entries of a general sort on philosophical topics germane to current discussions in psychoanalysis. Both authors and topics were left to my discretion. I thought the series was a good idea and gladly agreed to do it. To my surprise and pleasure, all the philosophers I invited accepted I am only sorry that the series could not be longer as there are other philosophers as well who would have been splendid participants, and other topics I would like to have addressed. The essays that will follow in subsequent issues represent by and large the tradition of analytic philosophy, though this has come in the last few decades to comprise many of the themes we used to associate with the Continental tradition. Future entries, by James Conant, Donald Davison, Pascal Engel, Dagfinn Follesdal, James Hopkins, Ernest Le Pore, Jeffrey Malpas, Jerome Neu, Brian O'Shaughnessy, Richard Rorty and Richard Wollheim, will address the following topics: intersubjectivity, meaning and language, consciousness and perception, pragmatism, knowledge and belief, norms and nature, metaphor, hermeneutics, truth, self-deception, the emotions. The essay below on knowledge, which will also be the topic of another entry by a different author later on, is the only one in the series that I will write. PMID- 10669972 TI - 'Getting in on the act: the hysterical solution'. PMID- 10669973 TI - 'The Piggle'. PMID- 10669974 TI - A neurocorrective approach for MMPI-2 use with brain-damaged patients. AB - Conventional administration of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) to aetiologically distinct brain-damaged out-patients (n = 137) revealed significant indications of psychological maladjustment. An adjustment for the endorsement of aetiology-specific items pertaining to traumatic brain injury (TBI), stroke, and whiplash was considered necessary, however, because these items may represent potentially valid symptoms or manifestations of neurological damage or dysfunction. These so-called neurologically relevant items (NRIs) were identified in a previous study. With this corrective approach, based on the complete MMPI-2 item pool, it was shown that T-score elevations could at least in part be attributed to symptoms associated with brain injury, regardless of the type of brain damage. Similarly, after prorated correction for the endorsement of NRIs, code-typing appeared to be substantially changed with respect to both occurrence and content of the MMPI-2 defined code-types. The validity of the NRI concept was supported by comparing NRI/non-NRI endorsement ratios of traumatically brain-injured patients with those of non-neurological patients, and with those having anxiety and somatoform disorders. To prevent unjustified interpretations when administering the MMPI-2 to brain-damaged patients, an adjustment procedure for NRI-endorsement is proposed, and difficulties in interpretation are discussed. PMID- 10669975 TI - Distribution of psychological aspects in subgroups of chronic low back pain patients divided on the score of physical performance. AB - This study investigates whether different subgroups of chronic low back pain patients (CLBPs) differ in psychological aspects assessed with the Symptom Checklist (SCL-90) and the Multidimensional Pain Questionnaire (MPI-DLV). Four subgroups of CLBPs are discerned using the results of lumbar dynamometry: 1. Patients with performances lower than healthy subjects (expected performance; n = 45). 2. Patients with performances comparable to healthy subjects (normal performance; n = 18). 3. Patients with inconsistent test behaviour (submaximal performance; n = 6). 4. Patients with performances that could be either maximal or submaximal (gray-zone performance; n = 10). Significant differences in psychological aspects were found between patients with submaximal and patients with expected performances but not between patients with normal and patients with expected performances. All patients with submaximal performance report a high degree of psychological distress, in contrast to 30% of those with normal performance and 20% of those with expected performance. Because of the differences found in psychological aspects between the CLBP subgroups, it is thought that a physical screening together with a psychological screening provides better insight in the two aspects of the deconditioning syndrome and thus can give better treatment indications than a physical screening alone. PMID- 10669976 TI - A multidimensional longitudinal analysis of family coping with brain injury. AB - Longitudinal research is needed to advance knowledge and practice in the area of family coping with brain injury. The purpose of this investigation was to examine the dimensions that underlie family coping with brain injury across a 2-year time period (1995 to 1997). Two-dimensional configurations of 30 family coping behaviours indicated a like family coping pattern of cognitive versus behavioural coping (Dimension 1) in both 1995 and 1997. The data also showed that families had a differing, or changed, family coping pattern along Dimension 2 from brain injury-focused coping versus family-to-community fit in 1995 to seeking professional help versus intra-family coping in 1997. Results thus indicated both changes and consistent coping patterns within the same group of families across time. Such information can be used to guide clinical family intervention, the development of long-term family support services, and future brain injury family research. PMID- 10669977 TI - Risk factors for disability pension among unemployed women on long-term sick leave. AB - BACKGROUND: In 1996 some 520,000 people (13% of Sweden's working force) were either long-term sick-listed or on disability pensions. To reintroduce sick and injured people to the workforce, vocational rehabilitation has received increasing emphasis. Unemployed women seem particularly difficult to rehabilitate. One explanation could be that unemployed women have more complex problems than others; another could concern the selection of cases for vocational rehabilitation programmes. AIM: The primary aim of this study was to test the hypothesis that unemployed women on sick-leave have more severe problems than others. A second purpose was to investigate whether the manner of selection for vocational rehabilitation programmes is different for unemployed women than for others. MATERIAL AND METHOD: The study analysed 364 registered long-term sick leave cases (90 days or more) initiated during 1992-1994 in the city of Stockholm, Sweden. RESULT: Our hypothesis was supported by the study. Unemployed women were exposed to more risk factors than unemployed men or employed men and women. Regarding the selection of cases for rehabilitation, no difference was present between unemployed women and others. A finding, however not statistically significant, was that people in vocational rehabilitation, regardless of sex and employment status, were less exposed to risk factors than people not undergoing rehabilitation. PMID- 10669978 TI - Collaborative partnerships in evaluation and experimental rehabilitation research. AB - Changes in rehabilitation and clinical practice, the recognition of the role of clients in achieving rehabilitation goals, and client participation in decision making and administration of service delivery have contributed to the development of new methodologies for research and evaluation. In this paper, the traditional relationship between researchers and subjects in traditional rehabilitation evaluation and research is contrasted with relationships between researchers and clients-consumers in newer methodologies. The limitations and challenges to these models are discussed. Collaborative research, characterized by shared power and non-hierarchical authority between client-consumers and researchers, is described as an alternative model for experimental and evaluative research. Collaborative relationships provide a structure for building upon the knowledge and expertise of each research partner. The authors draw upon their own experience of collaborative research partnerships and models of research partnerships are described. Research developed and conducted in a collaborative partnership can include quantitative and qualitative approaches, maintain traditional scientific perspectives of objectivity, reliability and replicability, and improve participation rates, continuity of involvement for longitudinal studies and utilization of researched methods into practice. PMID- 10669979 TI - Patients with metastatic breast cancer: their physical and psychological rehabilitation needs. AB - Few studies have monitored the physical and psychological sequelae of a diagnosis and treatment of metastatic breast cancer and the findings of the present study enables members of the rehabilitation team to understand the range of problems patients encounter, refer patients to other rehabilitation, and plan appropriate treatment. The present study aimed to describe the levels of anxiety and depression; to describe their rehabilitation status; and to ascertain whether any relationship exists between mood disturbance and physical rehabilitation status in a sample of women with metastatic breast cancer. Eighty patients with staging confirmed metastatic breast cancer were interviewed at home every eight weeks from diagnosis of metastases, on eight occasions using the Cancer Rehabilitation Evaluation System--Short Form, the Hospital Anxiety and Depression Scale, and an interview schedule to ascertain demographic and disease details. Results suggested that patients had a range of rehabilitation needs throughout the course of their disease. Mood disturbance was a significant problem in this sample of patients with many patients scoring in the case range for anxiety and depression on the HAD. A positive and strong relationship was found to exist between mood disturbance and physical rehabilitation status. The results of the study are discussed in the context of previous research and future research and clinical implications for the rehabilitation team are discussed. PMID- 10669980 TI - Psycho-social dysfunctions in patients after recovery from mania and depression. AB - Contrary to popular opinion, complete functional recovery does not occur in approximately 25% of patients with a diagnosis of mood disorders. The current study aimed at finding the recovery status in major mood disorders. A sample group of 122 patients (77 bipolar and 45 major depression) was selected from the outpatient department, fulfilling the DSM-IV diagnostic criteria. All patients had their index episodes at least one year prior to their date of inclusion and were either asymptomatic or mildly symptomatic during that time. Manics and depressives were rated with the Bech Raefelson Mania Scale (BRMS) and Hamilton Depression Rating Scale (HDRS) respectively. All the patients were also rated on the Brief Psychiatric Rating Scale (BPRS), Dysfunction Analysis Questionnaire (DAQ) and Global Evaluation Scale of Disability Assessment Schedule by WHO (GES/DAS). They were compared with 40 age and sex matched normal controls. It was found that the symptomatic recovery was better than the functional recovery in both manics and depressives and patients with major depression were marginally more dysfunctional compared to those with mania. It is concluded that a majority of patients of both mania and depression do not achieve complete functional recovery and are in need of on-going psychosocial rehabilitation. PMID- 10669981 TI - A survey of rehabilitative services and people coping with physical disabilities in Uganda, East Africa. AB - The impact of physical disability in the developing world is a tremendous health issue. The developing world presents many challenges in the lives of these people, many who cope without rehabilitative care. To determine how disabled people manage in this setting, 49 mobility-impaired Ugandans were interviewed. Functional capacity was assessed by determining the severity of lower extremity impairment and identifying the use of assistive devices and personal assistance needed in activities of daily living. Males appeared to have greater access to rehabilitative equipment, namely wheelchairs, and to personal assistance. The use of assistive devices was indicative of greater mobility, which subsequently provided greater opportunities, such as formal education or employment. Subjects with severe impairments without access to rehabilitative equipment were dependent upon crawling for mobility and begged for economic survival. Although there were a mix of responses towards perceived attitudes towards disabled subjects, more reported being viewed in a positive than in a negative light. Findings suggest that gender-related issues may limit access to rehabilitative equipment, due to limited financial resources of women living in the developing world. Uganda utilizes a viable political organization of people with disabilities to raise awareness and to empower its citizens coping with disability. PMID- 10669982 TI - Discharge to home among Hispanic and non-Hispanic stroke survivors: does family make a difference? PMID- 10669983 TI - Aberrant behaviour of persons with developmental disabilities as a function of the characteristics of training tasks. PMID- 10669985 TI - Social support in different communication environments. PMID- 10669984 TI - Outcome of obturator nerve block with alcohol for the treatment of hip adductor spasticity. PMID- 10669986 TI - Rehabilitation needs as a result of firearm-related injury; a four-year retrospective study from an inner-city hospital in the United States. PMID- 10669987 TI - Educational and behavioural characteristics of autistic children in the United Arab Emirates. PMID- 10669988 TI - Previous endoscopic treatment does not affect complication rate and outcome of laparoscopic Heller myotomy and anterior fundoplication for oesophageal achalasia. AB - BACKGROUND: Anedoctal reports suggest a detrimental effect of pneumatic dilation and botulinum toxin injection in patients who are potential candidates for Heller myotomy. AIMS: To assess symptomatic and objective outcome in patients undergoing Heller myotomy as a primary procedure or after failed endoscopic treatment. PATIENTS: Between November 1992 and December 1998, 92 patients with oesophageal achalasia were treated. Sixty patients had primary surgery; 32 patients had surgery after unsuccessful pneumatic dilation (n = 22), or botulinum toxin injection (n = 10). METHODS: Laparoscopic Heller myotomy plus Dor fundoplication with routine intraoperative endoscopy. Operative records, symptoms, and results of radiological, manometric and scintigraphic assessment in the two groups of patients were compared. RESULTS: The mean operative time, the rate of intraoperative mucosal tears and the incidence of postoperative dysphagia were similar in the two groups. Mucosal tears occurred more frequently during the first 30 operations (p < 0.05). Median follow-up was 28 months (range 4-76). An abnormal oesophageal acid exposure was documented in 2 patients in the primary surgery group (7.7%), and in 2 patients in the pneumatic dilation/botulinum toxin group (13.3%) (p = ns). Lower oesophageal sphincter pressure significantly decreased in both groups (p < 0.01). The mean percentage of radionuclide residual activity in the oesophagus at 1 and 10 minutes significantly decreased in both groups (p < 0.01). CONCLUSIONS: There was only a trend, although not statistically significant, towards an increased risk of complications and adverse effects in patients previously treated by pneumatic dilation or botulinum toxin. The higher incidence of mucosal tears during the first 30 operations suggests the effect of the learning curve. PMID- 10669989 TI - Helicobacter pylori eradication with dual and low-dose triple therapy in patients with liver cirrhosis. AB - BACKGROUND AND AIMS: Few data exist on the treatment of Helicobacter pylori infection in cirrhotic patients. In this study we assessed the efficacy of standard dual and one-week low-dose triple therapy on Helicobacter pylori eradication in cirrhotics. PATIENTS AND METHODS: In a prospective study, 83 cirrhotic patients with epigastric pain were randomised to receive either a two week course of dual therapy, composed of omeprazole 20 mg b.d. plus amoxycillin 1 g b.d. (n = 41) or a one-week course of triple therapy, composed of omeprazole 20 mg b.d., clarithromycin 250 mg b.d., and tetracycline 500 mg b.d (n = 42). Helicobacter pylori infection at entry and eradication 6-8 weeks after the end of therapy were assessed by rapid urease test and histology on biopsies from the antrum and corpus. When eradication did not occur with either dual or triple therapy, patients were given the alternative regimen. Helicobacter pylori eradication in these patients was assessed 6-8 weeks after the end of treatment by a further endoscopy. RESULTS: Helicobacter pylori eradication was achieved in 87.8% (36 out of 41; 95% confidence interval 77.8-97.8%) of patients after dual therapy and in 85.7% (36 out of 42; 95% confidence interval 75.1-96.3%) of patients treated with triple therapy (p = NS). In patients in whom initial eradication was unsuccessful, re-treatment eradicated Helicobacter pylori in 4 out of 5 patients given the triple regimen and in all 5 patients who received the dual therapy. One patient was lost to follow-up. No major side-effects were reported for either treatment regimen. CONCLUSIONS: Our data show that both dual and triple therapies are effective in Helicobacter pylori eradication in cirrhotics as well as in eradication failure patients. Therefore, the use of the dual therapy regimen is strongly suggested as an initial treatment for Helicobacter pylori eradication in cirrhotic patients. PMID- 10669990 TI - Differential features of gastric cancer patients, either Helicobacter pylori positive or Helicobacter pylori negative. AB - BACKGROUND: Helicobacter pylori infection is associated with an increased risk of gastric cancer. In Helicobacter pylori negative patients, factors different from those in Helicobacter pylori positive patients may be involved in gastric carcinogenesis. METHODS: Thirty-nine recently diagnosed consecutive patients with gastric cancer were investigated. Gastric biopsies were obtained for detection of Helicobacter pylori (by immunohistochemistry), non-Helicobacter pylori flora (by modified Giemsa and culture) and histological assessment according to the Sydney classification by Haematoxylin-Eosin staining. In serum samples, Helicobacter pylori antibodies were determined by IgG enzyme-linked immunosorbent assay, IgA enzyme-linked immunosorbent assay, and Western blotting. Furthermore, serum gastrin, pepsinogen A and C and plasma chromogranin A were determined. RESULTS: Helicobacter pylori was detected by immunohistochemistry in 53.8%, by IgG in 56.4%, by IgA in 33.3%, and by Western blotting in 74.4% of the 39 patients. Ten patients (25.6%) were negative by both histology and serology. Non-Helicobacter pylori flora was detected in 27 of the 39 patients (69.2%) with a similar frequency in Helicobacter pylori positive and negative patients. Helicobacter pylori positivity was found significantly more often in diffuse than intestinal type carcinoma patients (p < 0.05). Elevated gastrin levels and antrum-sparing atrophic gastritis were more frequent in Helicobacter pylori negative than in Helicobacter pylori positive patients (p < 0.05). CONCLUSIONS: In 10 out of 39 gastric cancer patients, no evidence of previous or current Helicobacter pylori infection could be demonstrated. Non-Helicobacter pylori was found in 69.2% of patients regardless of the Helicobacter pylori status. Further studies are needed to establish the contribution of non-Helicobacter pylori flora as well as antrum sparing atrophic gastritis with hypergastrinaemia to the development of gastric cancer. PMID- 10669991 TI - Is autoimmunity involved in the relationship between Helicobacter pylori infection, atrophic gastritis and gastric cancer? PMID- 10669992 TI - Macroamylase detection in serum using selective precipitation: a rapid and reliable assay. AB - BACKGROUND AND AIM: Available assays for measurement of pancreatic isoamylase in serum based on specific immunoinhibition of salivary fraction are unable to detect macroamylase. We combined a polyethylene glycol test which identifies macroamylase by selective precipitation with an automated assay for total amylase and pancreatic isoamylase measurement. METHODS: We analysed 24 sera proven positive for macroamylase using gel filtration chromatography and 80 negative sera. Precipitation of macroamylase with polyethylene glycol, colourimetric measurement of total amylase activity and immunoinhibition for the determination of pancreatic isoamylase were carried out. RESULTS: Macroamylasaemic sera showed precipitation of at least 71% of the amylase activity, while sera with normal sized amylase exhibited a maximum of 61%. In all the macroamylasaemic sera but two, the immunoinhibition test showed a rise in pancreatic isoamylase, which was found to be the prevalent fraction in 16. In 21 out of 24 sera with macroamylase and 67 out of 80 with normal-sized amylase, the precipitated amylase activity was also measured after immunoinhibition of non pancreatic activity. In macroamylasaemic sera, the percentage of precipitated pancreatic isoamylase activity ranged from 75% to 98%, while in samples with normal-sized amylase it was less than 71%. CONCLUSIONS: Polyethylene glycol precipitation can easily be combined with automated assays for the determination of pancreatic isoamylase and should be carried out whenever dealing with hyperamylasaemia of unclear origin. PMID- 10669993 TI - Clinical features and genotype-phenotype correlations in 41 Italian families with adenomatosis coli. AB - BACKGROUND: Familial Adenomatous Polyposis in an autosomal dominant disease in which the large bowel is carpeted by polyps of various dimensions appearing during the second or third decade of life. Several extracolonic manifestations complete the clinical spectrum of Familial Adenomatous Polyposis. If untreated, the disease leads invariably to colorectal cancer. The gene responsible for the disease, adenomatous Polyposis Coli, has been localized at chromosome 5q21. AIMS: To describe the clinical features of 156 Familial Adenomatous Polyposis patients (from 41 families) and to analyze possible correlations between genotype and phenotype. PATIENTS AND METHODS: Familial Adenomatous Polyposis was defined as the presence of 100 or more polyps in the large bowel. In 17 families (41%), the proband was the only affected individual (single cases). Adenomatous Polyposis Coli gene mutations were studied on DNA extracted from peripheral white blood cells and evaluated by polymerase chain reaction single strand conformation polymorphism, followed by direct sequencing of samples showing abnormal banding at single strand conformation polymorphism. RESULTS: The large majority of Familial Adenomatous Polyposis patients underwent surgery; colectomy with ileorectal anastomosis was the most frequent approach, however, cancer of the rectal stump developed in 11.6% of patients submitted to colectomy and ileorectal anastomosis. Adenomas were rare in the stomach (8.8%), but their frequency increased in the duodenum (33.8%) and jejunum (55.0%, chi 2 for trend 23.7, p < 0.001). Desmoid tumours were diagnosed in 17 patients (10.9% of the total) and in 6 families. Mutations of the Adenomatous Polyposis Coli gene were studied in 20 out of 25 families (80%) and on a total of 75 individuals. The most frequent alterations were 1 to 5 bp deletions leading to stop codons and truncated proteins. Desmoid tumours, presence of duodenal or jejunal adenomas were associated with an ample range of mutations, from codon 215 to codon 1464. In contrast, particularly severe polyposis (mean age at appearance of polyps 11-16 years, and of cancer development 27-32 years) was associated with a "hotspot" mutation site at codons 1303-1309. CONCLUSIONS: In patients with Familial Adenomatous Polyposis, subtotal colectomy with ileorectal anastomosis is still the treatment of choice. Adenomatous lesions seem to show a "gradient" distribution from the stomach to the large bowel. Desmoid tumours are relatively common, though their incidence is limited to some of the families. Constitutional mutations can be detected in 80% of the investigated families. Genotype-phenotype correlations showed a hot-spot at codons 1303-1309, frequently associated with severe polyposis. PMID- 10669994 TI - Circulating pro- and antioxidant factors in iron and porphyrin metabolism disorders. AB - BACKGROUND: Porphyria cutanea tarda and haemochromatosis are taken to be spontaneous human models of oxidative cellular damage, with an increased risk of fibrosis and cancer evolution. AIM: To define the relative pro-oxidant roles of porphyrin and iron, in their different molecular forms, and their effects on antioxidant biological systems. PATIENTS: A group of 17 patients with porphyria cutanea tarda and a group of 14 patients with primary and secondary haemochromatosis, were compared with 21 healthy controls. METHODS: Plasma retinol, tocopherol, alpha- and beta-carotene, ascorbic acid, glutathione, malonyldialdehyde and red blood cell free iron were determined using high performance liquid chromatography. RESULTS: Only a modest increase in iron stores was demonstrated in the porphyria cutanea tarda group; in the haemochromatosis patients ferritin levels were almost seven times higher. By contrast, there was a sharp and virtually identical increase in red blood cell free iron and malonyldialdehyde in both the patient groups. A significant reduction was observed in retinol, alpha-, beta-carotene and red blood cell glutathione levels being more marked in porphyria cutanea tarda than in haemochromatosis patients. CONCLUSIONS: The study confirms the strong pro-oxidant effects of porphyrins in vivo, through an induction of the free toxic iron form, even though the total iron pool is not greatly expanded. The additional free-iron and porphyrin oxidant effects are documented both in red blood cell and plasma in the porphyria cutanea tarda group. It confirmed that aging exerts a negative influence in terms of pro- and antioxidant balance in all cases, but particularly in the haemochromatosis group. PMID- 10669995 TI - Small bowel schwannoma with diffuse subcutaneous lipomatosis. Case report and literature review. AB - A case of a small-bowel schwannoma with diffuse familiar lipomatosis is described. This case underlines the rarity of the neoplasm and its probably chance association with subcutaneous lipomatosis. The intestinal neoplasm was diagnosed preoperatively by upper gastrointestinal endoscopy and a small-bowel enema; computed tomography scan confirmed the intestinal lesion. Attention is focused on the morphological features of intestinal schwannomas and their biological behaviour. PMID- 10669996 TI - Ischaemic necrotizing pancreatitis after cardiac surgery. A case report and review of the literature. AB - Ischaemia is a rare but often lethal aetiology of pancreatitis. A 67-year-old man underwent aortocoronary by-pass. Postoperatively, he developed atrial fibrillation and possibly acute myocardial infarction. Later, he had acute pancreatitis and underwent laparotomy for purulent peritonitis due to a ruptured pancreatic abscess. Cholesterolosis was found but no gallstones. The postoperative period was heavily complicated and the patient eventually died due to multiorgan failure. The occurrence of ischaemic pancreatitis should be more readily suspected in patients with abdominal symptoms following surgery that induces ischaemia of the pancreas. It is possible that delay in diagnosis accounts for the high death rate of such postoperative complication. PMID- 10669997 TI - Electrohydraulic lithotripsy treatment of gallstone after disimpaction of the stone from the duodenal bulb (Bouveret's syndrome). AB - A 75-year-old man with right upper quadrant abdominal pain was diagnosed by gastroscopy to have an impacted gallstone in the duodenal bulb. Using the polypectomy loop, the stone was extracted from the bulbus and mobilized into the stomach. After failure to remove the stone from the stomach as well as fragmentation by mechanical lithotripsy, electrohydraulic lithotripsy was used to break up the stone, parts of which passed spontaneously through the bowel. Thus, it was unnecessary to proceed with surgical enterolithotomy to remove, from the duodenal bulb, the impacted gallstone responsible for the gastric outlet obstruction. PMID- 10669998 TI - Current role of magnetic resonance cholangiopancreatography in the diagnosis of common bile duct and pancreatic diseases. AB - Magnetic resonance cholangiopancreatography is a noninvasive procedure that is increasingly used in patients with hepatobiliary and pancreatic diseases. The accuracy of magnetic resonance cholangiopancreatography has dramatically improved during the last few years, and there is no doubt that magnetic resonance cholangiopancreatography will have a major impact on the gastroenterologist's diagnostic work-up of patients. The key for success in dealing with hepatobiliary and pancreatic diseases is the precise knowledge of their course, the indications for treatment, and the therapy available. Thus, a team approach with strong representation from gastroenterology and radiology will be the most optimal path to an improved understanding of the value of Magnetic Resonance technology. PMID- 10670000 TI - Evolving concepts on inflammatory bowel disease. Are we happy with the present nosology? AB - The term inflammatory bowel disease traditionally comprises ulcerative colitis, Crohn's disease and indeterminate colitis, an intermediate variant of the two major forms. The term is commonly used in the literature and in clinical practice even though it has never been revised in a Consensus Conference. The present nosology of inflammatory bowel disease seems not to be entirely satisfactory as it is limited to chronic diseases only and does not include several recently described idiopathic inflammatory bowel disorders. Although the aetiology of inflammatory bowel disease remains unknown, both ulcerative colitis and Crohn's disease are characterized by a similar pathogenesis which consists in a persistent intestinal inflammation resulting from disregulation of the gut mucosal immune system. The pathogenetic mechanisms could, therefore, provide a suitable criterion for the classification of idiopathic inflammatory bowel disease. A revised classification of inflammatory bowel disease is thus proposed. It seems reasonable to subclassify inflammatory bowel disease into acute and chronic forms. Acute forms should include the sudden attacks of ulcerative colitis and Crohn's disease with rapid and complete resolution and the so-called "acute self-limited colitis". The chronic forms should comprise, besides the classical forms of ulcerative colitis, Crohn's disease and indeterminate colitis, also other idiopathic inflammatory bowel conditions such as collagenous colitis, lymphocytic colitis and eosinophilic gastroenteritis. PMID- 10669999 TI - Echinococcal liver cysts: treatment with echo-guided percutaneous puncture PAIR for echinococcal liver cysts. AB - In spite of recent progress, treatment for liver echinococcal cysts is still far from satisfying. In recent years, percutaneous drainage has been increasingly used for this purpose and it has been shown to be an effective alternative to surgery and chemotherapy alone. This technique is known as PAIR, from Puncture, Aspiration, Injection (of a scolecidal agent), Reaspiration: here we present our experience and the state of the art of PAIR. Patients from Italy and Turkana (Kenya), harbouring 233 Gharbi type I, II and III echinococcal cysts were successfully treated with PAIR: it was performed according to protocols established at the Division of Infectious and Tropical Diseases, IRCCS- Policlinico S. Matteo, University of Pavia. In Italy, one relapse was recorded, four years after the procedure; the patient was treated again with PAIR; no cases of anaphylactic shock or peritoneal dissemination were observed in a follow-up of 10 years; only 10 minor complications (biliary fistula, urticarioid reaction, abscessualization of the cyst, anaphylactoid reactions) were reported. Long-term results indicate that in Gharbi type I, II and III echinococcal cysts, and in developing countries, in particular, PAIR is a first choice method for the treatment of abdominal localizations of this disease. PMID- 10670001 TI - Renovascular impedance after total paracentesis. PMID- 10670002 TI - Conservative non-surgical management of congenital oesophageal stenosis associated with oesophageal atresia. PMID- 10670003 TI - AIDS care nursing: looking forward, thinking back. PMID- 10670004 TI - An investigation of the relationship between vitamin B12 deficiency and HIV infection. AB - The symptoms of vitamin B12 deficiency and several symptoms common to HIV infection overlap. Additionally, persons with HIV infection have frequently been found to have vitamin B12 deficiency. Therefore, the issue of concern is the prevalence of B12 deficiency in HIV-infected persons. A retrospective study of 63 medical records in a midwestern urban veterans affairs medical center provided the data for this study. Data were collected and analyzed for relationships among B12 levels, certain hematologic components, medications, symptomatology, and immune status. A chi-square goodness-of-fit test demonstrated a significant prevalence of B12 deficiency among persons with HIV disease, and chi-square two way tables demonstrated significant relationships between B12 deficiency and weight loss and diarrhea. Prevalence of B12 deficiency in persons with HIV infection has been shown to be significant, indicating that B12 deficiency screening in persons with HIV infection may need be done to aid the health care provider in planning the best possible care. Further research is needed to corroborate these findings. PMID- 10670005 TI - The Client Adherence Profiling-Intervention Tailoring (CAP-IT) intervention for enhancing adherence to HIV/AIDS medications: a pilot study. AB - This article describes the Client Adherence Profiling-Intervention Tailoring (CAP IT) intervention designed to enhance adherence to HIV/AIDS medications and reports the results of a pilot study aimed at assessing the feasibility of CAP IT. Initially, CAP-IT was designed to be implemented by nurse case managers during regularly scheduled home visits; it is currently under revision for use in an outpatient, ambulatory care setting. CAP-IT is an innovative, structured nursing assessment and care-planning activity that allows a standardized assessment of client needs and tailored highly active antiretroviral therapy adherence intervention strategies. CAP-IT is significantly different from the current standard nursing case management practice. Pilot study results in a sample of 10 home care patients suggests that clients have knowledge and skill deficits related to adherence and in the management of the side effects of medications. In addition, the pilot study supported the acceptability of the protocol to clients and the feasibility of integrating CAP-IT into nurse case manager practice. The pilot study results also provided evidence for the efficacy of CAP-IT. The next steps include testing CAP-IT in a randomized clinical trial to determine its effectiveness. PMID- 10670006 TI - A team approach to the treatment of AIDS wasting. AB - Despite the aggressive use of antiretroviral agents, AIDS wasting (AW) affects many persons infected with HIV. AW is characterized by a disproportionate loss of metabolically active tissue, specifically body cell mass--tissue involved with glucose oxidation, protein synthesis, and immune system function. AW correlates with poor quality of life and clinical outcomes. This condition requires a multidisciplinary team approach for effective management. Optimal maintenance of lean body mass and reversal of AW involves a combination of appropriate antiretroviral use, opportunistic infection prophylaxis, optimal nutrition, exercise, body composition monitoring, anabolic agents including growth hormone (rhGH[m]) and testosterone supplementation, mental health support, economic aid, and legal assistance. The team approach to treatment of AW requires the coordinated activity of nurses, dietitians, physicians, pharmacists, social workers, case managers, reimbursement personnel, caregiver(s), physical therapists, and the patient. This article, based on clinical experience treating AW, explains how the condition is managed using a multidisciplinary team approach. PMID- 10670007 TI - The utility of the Transtheoretical Model of behavior change for HIV risk reduction in injection drug users. AB - The spread of HIV among injection drug users (IDUs) is the second most common mode of transmission next to sexual contact. Although HIV infections can be prevented by changing high-risk behaviors such as needle sharing, these high-risk behaviors are highly complex. Initially developed for smoking cessation, Prochaska's Transtheoretical Model (TTM) is well-suited to the IDU population because it recognizes that chronic behavior patterns are usually under some combination of biological, social, and self-control. The objective of this article is to examine the utility of the TTM for promoting risk reduction behaviors among IDUs. This article will outline (a) the challenges of applying the TTM to IDU behaviors with respect to HIV prevention, (b) the four major components of the TTM as they relate to IDUs, (c) how risk reduction practitioners are currently using the TTM, and (d) current and future research using the TTM. PMID- 10670008 TI - The Transtheoretical Model and stress management practices in women at risk for, or infected with, HIV. AB - Although scientific inquiry using the Transtheoretical Model of Behavior (TTM) supports various behavior changes in multiple samples, no research was found that examined this model with women already infected with HIV. This article provides a brief overview of the related literature and describes a pilot study that evaluated TTM concepts in women at risk for, or infected with, HIV. The pilot study examined preliminary psychometrics of the research measures in women at risk for (n = 9), or infected with, HIV (n = 10), and examined predicted differences in situational confidence and stress management practices by HIV serostatus (positive vs. negative) and stage of change (precontemplation and contemplation vs. preparation, action, and maintenance) implied by the TTM. This pilot study supports use of the TTM to examine readiness to use stress management behavior in women regardless of their HIV serostatus. Further TTM stress management inquiry is encouraged to extend the knowledge base needed in caring for this vulnerable population. PMID- 10670009 TI - Immune reconstitution in the HAART era, Part 1: Immune abnormalities in HIV/AIDS. AB - Highly active antiretroviral therapy (HAART) has dramatically altered the course of HIV infection. HAART has been associated with a substantial decrease in HIV related morbidity and mortality and an improved quality of life for many persons living with HIV/AIDS. This improvement is due to suppression of viral replication and subsequent repletion of CD4+ T lymphocytes. However, it is not known if HAART can restore the deficits in immune function that are induced by HIV infection, such as anergy and disturbances in cellular phenotypes. In this first part of a two-part series, the immune abnormalities associated with HIV infection will be reviewed, and the concept of immune reconstitution will be introduced. In Part 2, the clinical significance of immune reconstitution will be discussed. PMID- 10670010 TI - Nevirapine prevents vertical HIV-1 transmission. PMID- 10670011 TI - Developing your idea for a research project. PMID- 10670012 TI - Program offers recognition of extraordinary partnerships in HIV care. PMID- 10670013 TI - Puncture-resistant gloves introduced. PMID- 10670014 TI - Life: past, present and future. AB - Molecular methods of taxonomy and phylogeny have changed the way in which life on earth is viewed; they have allowed us to transition from a eukaryote-centric (five-kingdoms) view of the planet to one that is peculiarly prokarote-centric, containing three kingdoms, two of which are prokaryotic unicells. These prokaryotes are distinguished from their eukaryotic counterparts by their toughness, tenacity and metabolic diversity. Realization of these features has, in many ways, changed the way we feel about life on earth, about the nature of life past and about the possibility of finding life elsewhere. In essence, the limits of life on this planet have expanded to such a degree that our thoughts of both past and future life have been altered. The abilities of prokaryotes to withstand many extreme conditions has led to the term extremophiles, used to describe the organisms that thrive under conditions thought just a few years ago, to be inconsistent with life. Perhaps the most extensive adaptation to extreme conditions, however, is represented by the ability of many bacteria to survive nutrient conditions not compatible with eukaryotic life. Prokaryotes have evolved to use nearly every redox couple that is in abundance on earth, filling the metabolic niches left behind by the oxygen-using, carbon-eating eukaryotes. This metabolic plasticity leads to a common feature in physically stratified environments of layered microbial communities, chemical indicators of the metabolic diversity of the prokaryotes. Such 'metabolic extremophily' forms a backdrop by which we can view the energy flow of life on this planet, think about what the evolutionary past of the planet might have been, and plan ways to look for life elsewhere, using the knowledge of energy flow on earth. PMID- 10670015 TI - Unanswered questions in ecology. AB - This is very much a personal view of what I think are some of the most important unanswered questions in ecology. That is, these are the questions that I expect will be high on the research agenda over the coming century. The list is organized hierarchically, beginning with questions at the level of individual populations, and progressing through interacting populations to entire communities or ecosystems. I will try to guess both at possible advances in basic knowledge and at potential applications. The only thing that is certain about this view of the future is that much of it will surely turn out to be wrong, and many of the most interesting future developments will be quite unforeseen. PMID- 10670016 TI - Theoretical biology in the third millennium. AB - During the 20th century our understanding of genetics and the processes of gene expression have undergone revolutionary change. Improved technology has identified the components of the living cell, and knowledge of the genetic code allows us to visualize the pathway from genotype to phenotype. We can now sequence entire genes, and improved cloning techniques enable us to transfer genes between organisms, giving a better understanding of their function. Due to the improved power of analytical tools databases of sequence information are growing at an exponential rate. Soon complete sequences of genomes and the three dimensional structure of all proteins may be known. The question we face in the new millennium is how to apply this data in a meaningful way. Since the genes carry the specification of an organism, and because they also record evolutionary changes, we need to design a theoretical framework that can take account of the flow of information through biological systems. PMID- 10670017 TI - Developmental biology and the redirection or replacement of cells. AB - The aim of developmental biology is to understand how an egg converts itself into a complete organism through the processes of cell differentiation, morphogenesis and size regulation. The principles that have emerged over recent decades include the constancy of the genome in nearly all cells of an individual, the existence of stem cells in many organs and the overwhelming importance of signalling between cells for the determination of their fate. These and other characteristics of development are discussed here in relation to the prospect of achieving cell and tissue correction or replacement with the help of nuclear transplantation and signalling factors. Nuclear transplantation offers a one-step procedure for generating multipotent embryo cells from the cells of an adult tissue such as skin. It should be possible to proliferate the resulting cells as can be done for mouse embryonic stem cells. Embryo cells can be made to differentiate in many directions by exposing them to various agents or to different concentrations of a single factor such as the transforming growth factor beta class signalling molecule activin. The possibility of a cancerous condition being acquired during these experimental manipulations can be guarded against by transfecting cells with a conditional suicide gene. Thus it may be possible to generate replacement cells or tissues from an adult human for transplantation back to the original donor, without the disadvantage of any genetic incompatibility. PMID- 10670018 TI - Structural biology. AB - Protein crystallography has become a major technique for understanding cellular processes. This has come about through great advances in the technology of data collection and interpretation, particularly the use of synchrotron radiation. The ability to express eukaryotic genes in Escherichia coli is also important. Analysis of known structures shows that all proteins are built from about 1000 primeval folds. The collection of all primeval folds provides a basis for predicting structure from sequence. At present about 450 are known. Of the presently sequenced genomes only a fraction can be related to known proteins on the basis of sequence alone. Attempts are being made to determine all (or as many as possible) of the structures from some bacterial genomes in the expectation that structure will point to function more reliably than does sequence. Membrane proteins present a special problem. The next 20 years may see the experimental determination of another 40,000 protein structures. This will make considerable demands on synchrotron sources and will require many more biochemists than are currently available. The availability of massive structure databases will alter the way biochemistry is done. PMID- 10670019 TI - Drugs for a new millennium. AB - A millennium, a century, even a decade is a long time-frame for speculation about anything. Advances in biomedical research in the last few decades have been so extraordinary and escalating at an ever-accelerating pace that any prophecy is a risky proposition. However, it is possible to divine the big, unanswered questions and envisage ways in which they might reasonably be approached in the next few decades, a task which I will try to essay. So many drugs treat so many different medical conditions that a detailed and comprehensive coverage would probably be tiresome. Instead, I will address certain broad themes and diseases that offer both immense challenges and great potential for advances. Rather than review detailed experimental issues, I will confine myself to the 'big picture' issues, providing examples of specific research only in a few instances drawing largely from areas I know best. PMID- 10670020 TI - From genotype to phenotype: genetics and medical practice in the new millennium. AB - The completion of the human genome project will provide a vast amount of information about human genetic diversity. One of the major challenges for the medical sciences will be to relate genotype to phenotype. Over recent years considerable progress has been made in relating the molecular pathology of monogenic diseases to the associated clinical phenotypes. Studies of the inherited disorders of haemoglobin, notably the thalassaemias, have shown how even in these, the simplest of monogenic diseases, there is remarkable complexity with respect to their phenotypic expression. Although studies of other monogenic diseases are less far advanced, it is clear that the same level of complexity will exist. This information provides some indication of the difficulties that will be met when trying to define the genes that are involved in common multigenic disorders and, in particular, in trying to relate disease phenotypes to the complex interactions between many genes and multiple environmental factors. PMID- 10670021 TI - Levels and loops: the future of artificial intelligence and neuroscience. AB - In discussing artificial intelligence and neuroscience, I will focus on two themes. The first is the universality of cycles (or loops): sets of variables that affect each other in such a way that any feed-forward account of causality and control, while informative, is misleading. The second theme is based around the observation that a computer is an intrinsically dualistic entity, with its physical set-up designed so as not to interfere with its logical set-up, which executes the computation. The brain is different. When analysed empirically at several different levels (cellular, molecular), it appears that there is no satisfactory way to separate a physical brain model (or algorithm, or representation), from a physical implementational substrate. When program and implementation are inseparable and thus interfere with each other, a dualistic point-of-view is impossible. Forced by empiricism into a monistic perspective, the brain-mind appears as neither embodied by or embedded in physical reality, but rather as identical to physical reality. This perspective has implications for the future of science and society. I will approach these from a negative point-of-view, by critiquing some of our millennial culture's popular projected futures. PMID- 10670022 TI - The impact of molecular biology on neuroscience. AB - How our brains work is one of the major unsolved problems of biology. This paper describes some of the techniques of molecular biology that are already being used to study the brains of animals. Mainly as a result of the human genome project many new techniques will soon become available which could decisively influence the progress of neuroscience. I suggest that neuroscientists should tell molecular biologists what their difficulties are, in the hope that this will stimulate the production of useful new biological tools. PMID- 10670024 TI - Splendours and miseries of the brain. AB - In this speculative essay, I examine two evolutionary developments underlying the enormous success of the human brain: its capacity to acquire knowledge and its variability across individuals. A feature of an efficient knowledge-acquiring system is, I believe, its capacity to abstract and to formulate ideals. Both attributes carry with them a clash between experience of the particular and what the brain has developed from experience of the many. Both therefore can lead to much disappointment in our daily lives. This disappointment is heightened by the fact that both abstraction and ideals are subject to variability in time within an individual and between individuals. Variability, which is a cherished source for evolutionary selection, can also be an isolating and individualizing feature in society. Thus the very features of the human brain which underlie our enormous evolutionary success can also be a major source of our misery. PMID- 10670025 TI - The future of philosophy. AB - There is no sharp dividing line between science and philosophy, but philosophical problems tend to have three special features. First, they tend to concern large frameworks rather than specific questions within the framework. Second, they are questions for which there is no generally accepted method of solution. And third they tend to involve conceptual issues. For these reasons a philosophical problem such as the nature of life can become a scientific problem if it is put into a shape where it admits of scientific resolution. Philosophy in the 20th century was characterized by a concern with logic and language, which is markedly different from the concerns of earlier centuries of philosophy. However, it shared with the European philosophical tradition since the 17th century an excessive concern with issues in the theory of knowledge and with scepticism. As the century ends, we can see that scepticism no longer occupies centre stage, and this enables us to have a more constructive approach to philosophical problems than was possible for earlier generations. This situation is somewhat analogous to the shift from the sceptical concerns of Socrates and Plato to the constructive philosophical enterprise of Aristotle. With that in mind, we can discuss the prospects for the following six philosophical areas: (1) the traditional mind-body problem; (ii) the philosophy of mind and cognitive science; (iii) the philosophy of language; (iv) the philosophy of society; (v) ethics and practical reasons; (vi) the philosophy of science. The general theme of these investigations, I believe, is that the appraisal of the true significance of issues in the philosophy of knowledge enables us to have a more constructive account of various other philosophical problems than has typically been possible for the past three centuries. PMID- 10670023 TI - The past, the future and the biology of memory storage. AB - We here briefly review a century of accomplishments in studying memory storage and delineate the two major questions that have dominated thinking in this area: the systems question of memory, which concerns where in the brain storage occurs; and the molecular question of memory, which concerns the mechanisms whereby memories are stored and maintained. We go on to consider the themes that memory research may be able to address in the 21st century. Finally, we reflect on the clinical and societal import of our increasing understanding of the mechanisms of memory, discussing possible therapeutic approaches to diseases that manifest with disruptions of learning and possible ethical implication of the ability, which is on the horizon, to ameliorate or even enhance human memory. PMID- 10670026 TI - Smell diskettes as screening test of olfaction. AB - A screening test of olfaction was developed with reusable diskettes as applicators of 8 different odorants. Using a questionnaire with illustrations, the test was designed as a triple forced multiple choice test resulting in a score of 0 to 8 correct answers. To validate the test, 102 volunteers with normal olfaction, as well as 22 patients with subjective hyposmia or anosmia, were tested. To compare the developed test with an already validated method, the same persons also performed the sniffin' sticks screening test. The results indicate that the screening test with smell diskettes recognizes patients with normal olfaction and consistently distinguishes them from patients with hyposmia or anosmia. PMID- 10670027 TI - A new approach to improving illumination in the nose during endonasal surgery. AB - Despite improvements in light sources the problem of illumination during endonasal surgery persists. This is particularly so in the presence of blood which absorbs light and renders the operative field dark as a consequence. This paper describes a series of in vitro experiments that show how improved illumination is possible using readily available, inexpensive, sterilisable and flexible materials. The hypothesis tested was that white coloured materials, when placed into the nasal cavity during endonasal surgery, improve illumination of the operative field by reflecting light onto the area of surgical interest. This hypothesis was tested with the use of a light proof box into which were introduced blood coloured and reflective materials. The light reflected back from a fixed blood coloured surface within the box was measured. The introduction of white materials into the box provided greater illumination than blue or foil surfaces. PMID- 10670028 TI - Management of sinonasal hemangiopericytomas. AB - The purpose of the present study is to report four cases of sinonasal hemangiopericytoma (HP) diagnosed and treated in our department between 1987 and 1998. The pretreatment findings and the treatment are described and discussed in the light of the literature. HP are unusual vascular tumors, featuring pericytes distributed around normal vascular channels. Two of these four cases were located in the nasal cavity and the other two were located in the maxillary sinus. Inside the nasal cavity, HP presented as a protruding reddish-gray mass with marked bleeding on contact. Electron microscopy and immunohistochemical techniques are essential for diagnosis and to distinguish HP from other sarcomatous tumors. Preoperative assessment included routine CT, MRI, arteriography and selective embolization. These tumors must be treated surgically with complete excision. An endonasal approach was performed in two cases of intranasal HP, while a combined external-endonasal approach was required for the other two cases of HP. PMID- 10670029 TI - p53 over-expression and its correlation with PCNA index in nasal polyps. AB - Our knowledge about the etiopathogenesis of nasal polyps (Nps) is still limited. In this study, in order to define the biological features of these neoformations, we investigated with immunohistochemistry the p53 over-expression and the proliferating cell nuclear antigen (PCNA) in 32 cases of Nps and in normal mucosa of 11 control cases. The evaluation of PCNA showed a wide range of indices (0.5 18.2%) with a mean value (6.8%) significantly higher than in normal mucosa (2.9%). Over-expression of the p53 oncoprotein, observed in 50% of Nps, was statistically related to a high PCNA-index (> 6.8%). Our results suggest that Nps can behave, in a high percentage of cases, like tumours. PMID- 10670030 TI - How to manage patients with hard-to-recognize postnasal drip? AB - Postnasal drip (PND), commonly regarded as a phenomenon wherein nasal fluid drips into the pharynx, is one of the main symptoms of chronic sinusitis and other nasal lesions. This definition is controversial, however, because some patients have PND with no evidence of fluid either in the oropharynx or around the choanae. Among 220 patients in whom PND was diagnosed at the University of Tokyo Hospital between January 1996 and December 1997, 19 (8.6%) had hard-to-recognize PND on routine examination. Through careful observation, responsible lesions could be identified in all patients with hard-to-recognize PND. Seven patients had latent chronic sinusitis, 5 had nasopharyngeal lesions, such as Tornwaldt's cyst and inflammation, 3 had "old man's PND", 2 had "reflux PND", and 2 had polyps around the sphenoid ostium. Five patients received no treatment. In the other patients, medical therapy, mainly long-term treatment with low doses of macrolides, now regarded as a standard regimen for intractable chronic sinusitis in Japan, was effective in alleviating symptoms regardless of the cause. PMID- 10670031 TI - Non-specific nasal provocation test with histamine. Analysis of the dose-response curve. AB - Non-Specific Nasal Hyper-reactivity (NSNH) is described as a clinical condition characterized by the presence of rhinitic symptoms that are a consequence of non specific stimulations. Because of its effects on vascular, epithelial, and glandular receptors, NSNP Test (NSNPT) with histamine allows the study of NSNH. The aims of this study are 1. to analyze the behavior of NSNH both in non allergic chronic vasomotory patients and in healthy control subjects 2. to correlate total nasal resistance(TNR) to each dosage of histamine to derive the dose/response curves and 3. to study these curves to analyze and possibly define different stages according to the intensities of response of NSNH. We have studied 26 subjects affected by non-allergic vasomotor rhinitis and 10 healthy control subjects. We sprayed a NSNPT with histamine-phosphate (0.2-0.3-0.4-0.5 0.6-0.8 mg) in different sessions to avoid accumulation phenomena. Five minutes before and five minutes after each challenge, TNR was determined by active anterior rhinomanometry. TNR was correlated to the doses of histamine by an empirical equation. The most important results of this study are as follows: a) the variation of TNR follows a model of exponential curve, b) it is possible to classify NSNH, as a function of the regression b coefficient belonging to the empirical equation used, in reactivity classes, c) from one reactivity class to another, post-stimulation TNRs double; 0.5 mg of histamine of the NSNPT is the optimal dose, d) there is an overlap between the responses of some normal subjects and rhinopathic patients that will be the subject of a further study. Finally, our data suggest that, in a future perspective, it is possible to use the NSNPT with histamine for diagnostic, prognostic and therapeutic control purposes. PMID- 10670032 TI - Experimentally induced nasal irritation. AB - The aim of this study was to develop a method that is suited for the induction of nasal irritation. For this purpose inflammatory responses were analysed after challenging the nasal mucosa with experimentally induced cold, dry air (8 l/min, 22 degrees C, 20 %RH). To assess inflammatory effects we determined inflammatory mediators (prostaglandin E2 [PGE2], thromboxane B2 TXB2[, peptide leukotrienes pLT: LTC4, LTD4, LTE4[) in nasal lavage fluid which was sampled before, immediately after suprathreshold stimulation, and one hour after termination of the stimulation. In addition, subjects estimated the intensity of pain during the stimulation. Cold, dry air produced strong painful sensations which increased throughout the stimulation period. A significant increase of the inflammatory mediator pLT was observed after stimulation; mean concentrations of PGE2 and TXB2 also showed a tendency to increase. One hour after termination of the stimulation the concentration of these inflammatory mediators returned to baseline which indicates the reversibility of the effects of nasal irritation. These data suggest, that this model may be a useful tool in investigations of mucosal irritation as, for example, induced by environmental agents. PMID- 10670033 TI - Technical problems with protein extraction of chemokines featuring RANTES. AB - Chemokines are known to be one of the sources for eosinophilic tissue infiltration in eosinophilic inflammation. Detection of beta-chemokines such as RANTES was possible in nasal tissue with or without eosinophilic infiltration. The concentration of chemokines which has been measured in the same tissue differs often in the literature. Aim of this study was to compare the different techniques of protein extraction and help to understand and interpret the investigation on RANTES secretion. Tissue of nasal polyps, inferior and middle turbinate was cut into halves and every half on its own pulverized using liquid nitrogen. The protein extraction was performed either with citric acid solution (pH 2.5) or phosphate buffered saline (PBS). The samples were then lyophilized. The concentration of RANTES was measured by a specific double sandwich ELISA. Using the citric acid technique the average concentration of RANTES in middle turbinates was 1.3 ng/mg, in inferior turbinates 1.6 ng/mg and in polyps 2.6 ng/mg tissue, using the PBS technique respectively 0.6 ng/mg, 0.5 ng/mg and 0.8 ng/mg tissue. Our data revealed a mismatch of 3.3:1 for polyps (citric acid: PBS), 3.2:1 for inferior and 2.2:1 for middle turbinates, respectively. Consequent comparison between the results of different techniques was not possible. Of special interest was also the fact that different techniques had different efficiencies of protein extraction in different tissues. Present statements on RANTES concentrations as a prognostic factor in nasal tissues need a technically careful standardization as far as this study shows. PMID- 10670034 TI - An algorithm for the management of CSF rhinorrhoea illustrated by 36 cases. AB - The diagnosis of cerebrospinal fluid (CSF) rhinorrhoea should be established beyond reasonable doubt before surgical intervention is embarked upon. It is important not to miss the diagnosis in view of the real potential complication of meningitis if it is left untreated. We describe a management algorithm which centers around the selective use of nasal endoscopy, immunofixation of beta2 transferrin, high resolution coronal CT scans, and fluorescein lumbar puncture. This management strategy is illustrated with 36 cases. We have developed a minimally invasive endoscopic technique to repair CSF leaks, and in 30 patients we had a success rate of 93% after one procedure. PMID- 10670035 TI - Intrasphenoidal encephalocele and spontaneous CSF rhinorrhoea. AB - Intrasphenoidal encephalocele is a rare clinical entity. In the international literature only 16 cases have been reported up today, with female predominance. Clinically they manifest at middle and advanced ages (40-67 years), when spontaneous CSF rhinorrhoea or recurrent meningitis occurs. We present our case, a 46 years old female, who had CSF rhinorrhoea from the right vestibule for 10 months. The diagnosis was based on the history and the high-resolution brain and skull base CT-scanning in conjunction with opaque fluid injection in the subarachnoidal space through a lumbar puncture. She was successfully treated with an operation, through an endonasal trans-ethmoid microendoscopic approach, using the Draf and Stammberger technique. We discuss the pathogenesis of the intrasphenoidal encephalocele, the existence of small occult defects in the skull base, which cause, at the middle and advanced ages, CSF fistula with spontaneous CSF rhinorrhoea and/or recurrent meningitis. Finally we emphasize the advantages of the endonasal surgical approach for the treatment of this condition. PMID- 10670036 TI - Paranasal mucous cyst: a rare finding following septorhinoplasty. AB - Postoperative niucous cysts of the facial soft tissue are a rare complication after septorhinoplasty. We present a case of postseptorhinoplasty mucous cyst with a paranasal localisation. According to the literature available to us this localisation is extremely rare and has not been described before. Aetiology and possibilities to decrease the risk of such complications are discussed. PMID- 10670037 TI - [New recommendations for the treatment of the hypertensive patient: a theoretical concept or a useful tool in practice?]. PMID- 10670038 TI - [Highlights of rheumatology 1999: new developments between immunology and pain of movement mechanisms]. PMID- 10670039 TI - [The craft of robotic orthopedic surgery and Y2K]. PMID- 10670040 TI - [Advances in urology: the responsibility of family practice development]. PMID- 10670041 TI - [Oncology 1999: advances in immunotherapy]. PMID- 10670042 TI - [Medical treatment of heart failure: old facts in new lights]. PMID- 10670043 TI - [Hematology: clarity on the 75th birthday--a start in the new year with new ideas]. PMID- 10670044 TI - [Nephrology 1999: substantial Swiss contributions]. PMID- 10670045 TI - [Gynecology 1999]. PMID- 10670046 TI - [Hepatitis B: when to use lamivudine?]. PMID- 10670047 TI - [Arterioportal fistula as a cause of esophageal varices bleeding]. PMID- 10670048 TI - Imaging of middle ear pathology. PMID- 10670049 TI - Imaging of salivary gland disease. PMID- 10670050 TI - Oral cavity, oropharynx, and hypopharynx. PMID- 10670051 TI - Imaging of the larynx. PMID- 10670052 TI - Imaging of lymph nodes in the neck. PMID- 10670053 TI - Appropriate use of ultrasonography in the neck. PMID- 10670054 TI - Suprahyoid spaces of the head and neck. PMID- 10670055 TI - Applications of interventional neuroradiology in the head and neck. PMID- 10670056 TI - Imaging of postoperative larynx and neck. PMID- 10670057 TI - Post-therapeutic imaging of upper aerodigestive tract tumors. PMID- 10670058 TI - [Mobility of the back of the foot and myoelectric activity pattern in jumping exercises under increased stretching]. PMID- 10670059 TI - [Achilles tendon rupture: epidemiology, etiology, diagnosis and current treatment possibilities]. AB - Treatment of Achilles tendon rupture is up to date not standardized. Open surgical repair in different techniques, percutaneous repair and conservative treatment is controversial in discussion. The following paper shows the different techniques used today in Achilles tendon rupture. Results are represented and discussed. Followed by a recommendation of treatment in acute Achilles tendon ruptures. PMID- 10670060 TI - [Muscle strength after surgical treatment of ruptures of the long biceps tendon by refixation to the short head]. AB - Operative treatment for ruptures of the long biceps tendon still is discussed controversially. In the present literature surgical repair is advised for sporting and high demand patients due to favourable functional results and low loss of strength. Until now only few objective and reproducible results were published. In the present literature the keyhole-technique is recommended due to favourable bio-mechanical conditions. To evaluate postoperative strength 19 patients were investigated after an average follow-up of 6.5 years, by clinical examination and isokinetic measurement. Compared with the non-operated shoulder isokinetic determination of isometric maximal peak torque and strength during concentric stress for elbow-flexion, shoulder-abduction and shoulder-flexion yield to almost identical results for the operated shoulder. According to the criteria of the Constant-Score all patients achieved very good and good results. Refixation to the short head can be advised for treatment of ruptures of the long biceps tendon due to the certain technique with a low complication rate and very good functional outcome. PMID- 10670061 TI - [Years of training: a new risk factor in acute badminton injuries]. AB - The incidence of badminton injuries is low compared with other sports, but acute injuries are generally more severe. Little is known about the risk of competitive badminton players to get an acute badminton injury. The purpose of this study was to define for the first time "years of playing badminton on a competitive level" as a risk factor for acute badminton injuries. 179 badminton injuries of 102 Austrian competitive badminton players, some of them being elite players in the european championships, were retrospectively registered. The years 1993, 1994 and 1995 were covered by our investigation. Injury incidences were defined as injuries/1000 h and were calculated separately for the 0. through the 21st year of competitive badminton. The incidence of acute badminton injuries increased constantly from the 0 to the 7th year of competitive badminton. The 6th and 7th year of competitive badminton showed a threefold increased incidence of acute badminton injuries when compared with the first year and the late years of the career as a competitive badminton player. We found no correlation between the incidence of acute badminton injuries and age, gender, hours played per year or time of warming up. We conclude that players being engaged in competitive badminton for 5 to 8 years represent a high risk population for acute badminton injuries. In contrast players at the beginning and towards the end of the competitive career showed a markedly lower risk for acute badminton injuries. We recommend additional training efforts concerning technical skills and endurance as well as proper rehabilitation of sports injuries for the high risk group of badminton players. PMID- 10670062 TI - [Injury and exertion patterns in football on artificial turf]. AB - This controlled non-selected cross-sectional study supplies a basic survey on the topic analysing 1783 injuries in 433 of 736 athletes out of a closed collective. Aged 11 to 40 years and having played an average of 3.7 years an artificial turf the players had sustained 38% skin injuries (58% in the legs), 28% sprains (64% in the ankle joints, 21% in the knee joints) and 17% muscle injuries. 76% of all injuries were minor, i.e. leading to an interruption of under one week, only 8% were severe with a break of over 3 weeks. The average risk of injury was 6 per 1000 hours of participation, similar to that in football on natural grass. More than half of the players protocol pain in the joints, muscles or column persisting even one day after the game, which only led to medical assistance in 3% of all cases. Playing football on artificial grass displays a specific pattern of injuries and exertion syndromes without a higher rate or grade of injuries and therefore shows no medical need for restriction. PMID- 10670063 TI - [Injury mechanisms in windsurfing regatta]. AB - In a retrospective study, we evaluated the injuries of 44 semi-professional competitors for the German Windsurf Cup, which were suffered from during one windsurfing season. This Cup is the national qualification tour for the annual "production fun board world championship". The subjects, participating in our study were randomly chosen. There were no surf-specific differences between the two groups. The average age was 24.63% had competitive surfing as their hobby, 37% were professional or semi-professional board sailors. The subjects surfed an average of 85 days in 1995. 23 (52%) windsurfers did not get hurt during the entire season. 21 (48%) of them got injured during the 1995 windsurf season. This is an incidence of only one injury per 174 windsurfing days. Only three windsurfers were injured during a competition. The other 18 occurred during training sessions. Most accidents happened because of an overpower situation, i.e. the sail was too big for the wind force (43%), or through negligence on the part of the windsurfer (19%). The most frequent type of the accident was the so called catapult crash (57%). The most common injuries were ligament ruptures of the lower leg (33%) and head burst wounds (19%). Compared with other competitive fun sports (e.g. snow boarding), windsurfing has a lower injury risk. In regard to the injury mechanisms, prophylactic recommendations are made. PMID- 10670064 TI - [Extra-articular localization of nodular synovitis: a case report]. AB - INTRODUCTION: The extra-articular nodular synovitis is regarded as a rare case of the common pigmented villonodular synovitis. The following report points out a case with wide extension in a young man's proximal lower leg. CASE-REPORT: A 24 year-old man complained about a non-painful tumor at his left proximal lower leg without any clinical handicap. Some years before he got a fracture of the proximal left tibia caused by an adequate sports trauma. So he had to undergo two operative investigations. The radiographs showed a transparent soft tissue tumor, which displaced the corticalis of the bone for some centimeters. The magnetic resonance imaging did not hint at a malignant process. After operative treatment the diagnosis was confirmed by histology. CONCLUSION: By operative and histological investigation advice could given to a benign tumor. The etiology of the extra-articular nodular synovitis is to be regarded as unknown. In our case a dislocation of synovial cells to extra-articular should be discussed. PMID- 10670065 TI - [Innovations in technique and use of ultrasound]. PMID- 10670066 TI - [Medical time requirements for abdominal ultrasound diagnosis]. PMID- 10670067 TI - [Ultrasound-guided transthoracic puncture]. AB - The diagnosis of peripheral lung foci may prove difficult. In addition to transthoracic puncture under X-ray fluoroscopy or CT control, ultrasound-guided puncture was shown to be a useful alternative. A prerequisite, however, is that the lesion should extend up to the pleura. This overview covers 97 original papers, of which 26 mainly consisted of lung punctures in a total of 1876 patients. The accuracy in carcinomas and metastases was 70 to 97%, on average markedly higher than 90%. Benign lesions are histologically more difficult to distinguish; here the accuracy is 70%. Partly due to pre-selection the method has a very low rate of complications. The rate of pneumothorax is 2.6%, those requiring drainage are about 1%. Haemoptyses occur 1-2% of the punctures, most commonly in cases of chronic pneumonia; colour-coded Duplex sonography is especially recommended in these cases because of the strong and regular vascularisation. The rate of complication increases in direct proportion to needle thickness. The possibilities of ultrasound-guided lung abscess drainage are also discussed. An intrathoracic lesion that is accessible to ultrasound imaging should be punctured today under ultrasound guidance, as this procedure is minimally stressful for the patient, is accurate, has a low rate of complications and is also cost effective. PMID- 10670068 TI - [Project graph technique for time management in abdominal ultrasound evaluations]. AB - PURPOSE: German insurance companies are cutting down the time required for ultrasound examinations. To determine the minimal examination time to perform an ultrasound examination a project graph technique was applied. MATERIALS AND METHODS: Time measurements of abdominal ultrasound examinations were performed by two independent observers. The different jobs for the performance of an ultrasound examination were determined and the critical pathway method applied. The total available time for abdominal ultrasound examinations (leeway) was determined, the minimal time to perform each job was measured and the critical time required for the procedure was calculated. RESULTS: 14 different jobs were identified to complete one abdominal ultrasound examination. The project graph displayed the shortest possible time of 24 minutes to perform an ultrasound examination. The pure ultrasound exam without colour Doppler examination was 6 minutes ("hands on the ultrasound probe"). The jobs performed by the physician were fully within the critical period. In consequence, the physician had no leeway or time lag in relation to a total time of 24 minutes for an ultrasound examination, whereas by contrast the nurse has a total leeway of 7.5 minutes. CONCLUSIONS: The applied project graph technique is an effective instrument for the purpose of quality management for hospitals as well as in private practice. The workflow and actions necessary to perform a treatment or examination can be analysed. Human resources management and cost planning should be performed on the basis of project graphs. PMID- 10670069 TI - [Intestinal B-mode sonography in patients with endemic sprue. Intestinal sonography in endemic sprue]. AB - AIM: The value of ultrasonography in the diagnosis, follow-up and for the detection of complications in patients with celiac sprue has not yet been sufficiently evaluated. A pronounced back and forth motility with echo-rich hump reflexes in a fluid-filled small bowel with a reduction of Kerckring's plicae circulares and with a loss of their density and uniformity was empirically defined as a diagnostic sign of celiac sprue. In the present study, the sonographic signs of celiac sprue were examined as an indicator of active sprue. METHOD: 50 patients with histologically proven celiac sprue were examined with real time ultrasonography (3.5-7 MHz). The detection or exclusion of the defined sonographic signs of celiac sprue with intensified motility and reduction of Kerckring's plicae circulares with a loss of their density and uniformity were evaluated by two independent examiners and documented without knowledge of the clinical findings. The clinical activity (active vs. remission) was assessed according to clinical criteria (diarrhea, steatorrhea, weight loss). 38 healthy subjects and 50 patients with Crohn's disease served as controls. RESULTS: In all 138 patients and controls adequate visualization of the bowel was achieved. In 16/50 (32%) patients with active celiac sprue changes of motility and reduction of Kerckring's plicae circulares with a loss of their density and uniformity were detected, whereas all 34/50 (68%) of patients with celiac sprue in remission did not have this pattern. In none of the controls with Crohn's disease or in the healthy subjects comparative sonographic signs of active celiac sprue were observed. In four patients with active celiac sprue a circumscript echopoor tumor of the small bowel wall could be sonographically detected, which turned out to be T-cell lymphoma in three and a carcinoma of the small intestine in one patient. An increased number of and/or enlarged mesenteric lymph nodes were found in patients with active celiac sprue. CONCLUSION: Changes of motility and reduction of Kerckring's plicae circulares with loss of density and uniformity at ultrasonography are a reliable indicator of active celiac sprue. PMID- 10670070 TI - [A test object for quality control of the instrument for doppler (duplex) ultrasonography, based on the Draft IEC 61685 Standard]. AB - PURPOSE: The authors, forming part of a multicenter project funded by the European Community, summarize the validation of a tissue-mimicking flow Doppler test object and of procedures for testing medical diagnostic Doppler equipment. The results of the project are expected to contribute to a future international IEC Standard concerning flow Doppler test objects (Draft IEC 61685 Standard) and for the European Medical Device Directive (MD 93/42/EEC). METHODS: Within this project a test protocol was developed that includes a set of different procedures, suitable for checking Spectral and Colour Doppler systems. The performance parameters for describing the image quality as well as the accuracy and the correct functioning of a system are in accordance with the definitions made in the Draft IEC 61685 Standard. RESULTS: A survey of the design and materials used for this Doppler test object will be presented with a special emphasis on the suitability of the procedures for routine measurement of performance parameters in hospitals. CONCLUSION: The test object satisfies the requirements of the Draft IEC 61685 Standard. The test procedures in combination with this test object can be used for checking different transducer models with nominal frequencies between 2.5-10.0 MHz. PMID- 10670071 TI - [Deep vein thrombosis following total arthroplasty hip replacement. Ultrasonographic screening in the rehabilitation phase]. AB - Deep vein thrombosis requires intensive therapy--either the patients require hospitalisation or their rehabilitation scheme must be adjusted and medical treatment applied. The total number of patients with deep vein thrombosis undetected before admission to rehabilitation facilities is unknown. AIM: Study parameters: 1. The occurrence of undetected deep vein thrombosis upon admittance to a rehabilitation facility. 2. The value of sonography in diagnosing, localising and determining the extent of deep vein thrombosis and 3. the influence of prophylactic antithrombotic treatment applied in the surgical hospital, as well as during the intermittent outpatient phase before admittance to the rehabilitation centre, on the occurrence of deep vein thrombosis. METHODS: 305 patients were examined between Nov. 95 and Dec. 96. The first check was carried out on the day of admission (on the 26th postoperative day, on average). The second check was done on the day of discharge from the rehabilitation centre. RESULTS: Deep vein thrombosis was found in 30 patients (10%). In 27 cases the thrombosis was located in the thigh, in one case it was found in the contralateral leg. 18 of these patients had been discharged from the surgical hospital on the 22nd postoperative day, for an average time of 7 days of outpatient care, before being admitted to the rehabilitation centre. 11 of these patients had not received any prophylactic antithrombotic treatment. One patient developed deep vein thrombosis of the thigh during the rehabilitation phase despite of prophylactic treatment. CONCLUSION: Postoperative sonographic screening of all patients undergoing hip surgery is a useful method to detect deep vein thrombosis. Prophylactic postoperative antithrombotic treatment in the rehabilitation phase is mandatory. PMID- 10670072 TI - [Clinical manifestations of schizencephaly and its sonographic diagnosis]. AB - Schizencephaly is defined as a cerebral malformation of the CNS with various clefts of the cerebral cortex. We report on two patients referred to our department with neurological abnormalities. In both cases the cranial sonography already provided for the clinical picture of schizencephaly. A MR-scan confirmed the diagnosis. In addition one of the patients proved to have a migrational disorder. The analysis of these cases and the relevant literature point out how difficult the etiologic differentiation is and, on the other hand, how various the manifestations of the malformation can be. The important role of cranial sonography as a screening method is shown. PMID- 10670073 TI - Pitfall: a pseudo tumor within the left liver lobe presenting with abdominal pain, jaundice and severe weight loss. AB - A 51 year old male patient with a history of chronic alcohol consumption and recurrent pancreatitis was referred to our hospital with jaundice, epigastric pain, severe diarrhoea and weight loss of 28 kg within the last 12 months. A CT scan of the abdomen 4 months before admission had shown a pancreatitis with free fluid around the corpus and tail of the pancreas as well as dilated intrahepatic bile ducts and a cavernous transformation of the portal vein. Moreover, a tumor (3.5 x 3.0 x 3.6 cm) with irregular contrast enhancement was seen within the left liver lobe. The patient was referred to us for further evaluation and treatment. The initial B-Mode sonogram revealed a bull's eye like well defined lesion (8.1 x 7.5 x 7.0 cm) within the left liver lobe, consistent with a tumour or abscess. Prior to a diagnostic needle biopsy a PTCD was performed in this case presenting with dilated intrahepatic bile ducts and having a history of Billroth II operation. An additional colour coded Duplex Doppler ultrasonography demonstrated a visceral artery aneurysm and prevented us from performing the diagnostic puncture. The aneurysm was assumed to originate from a variant or a branch of the left hepatic artery. Angiography revealed a pseudoaneurysm of the pancreaticoduodenal artery and coil embolization was performed because of the increasing size and the risk of a bleeding complication. Postinterventional colour duplex ultrasound measurement showed no blood flow within the aneurysm. Retrospectively, the pseudoaneurysm must have led to a compression of the common bile duct, since the patient did not develop cholestasis after embolization and removal of the PTCD. Thus, a pseudoaneurysm of the pancreaticoduodenal artery must be included in the differential diagnosis of liver tumours in patients with chronic pancreatitis, despite its unusual localization near the liver. Therefore, we suggest that colour coded ultrasonography should be applied to any unclear, bull's eye like lesion, even though this method alone cannot exactly determine the origin of the pseudoaneurysm. Interventional angiography remains the gold standard for the diagnosis and therapy of visceral artery aneurysm. PMID- 10670074 TI - [On the article: Bock, U., Mollhoff, T., Forster, R.: Ultrasonography guided versus anatomically oriented puncture of the internal jugular vein for central venous catheterization]. PMID- 10670075 TI - Adaptation of pregnant ewes to an exclusive onion diet. AB - A diet consisting entirely of cull onions fed to pregnant ewes produced Heinz body hemolytic anemia in all sheep after 21 d. After 28 d of daily consumption of 20 kg of onions/ewe, the anemia stabilized, and for the remaining 74 d the packed cell volume increased in the majority of sheep, although it did not return to normal. Compared to control ewes fed an alfalfa and grain diet, the onion-fed ewes had comparable body condition scores and fleece weights. There was no significant difference (alpha = 0.05) in pregnancy or lambing rate, number of lambs born/ewe exposed, or number of lambs born/ewe lambing. Greater numbers of sulfate-reducing bacteria (Desulfovibrio spp) and more ruminal hydrogen sulfide were present in onion-fed sheep compared to controls. Although an average 27% reduction in packed cell volume and Heinz body anemia developed in the onion-fed ewes, on the basis of this study it appears that pregnant ewes may be fed a pure onion diet with minimal detrimental effects. This adaptation to a pure onion diet is in part likely due to the apparent ability of the sheep's rumen to quickly develop a population of sulfate-reducing bacteria that decrease the toxicity of onion disulfides. PMID- 10670076 TI - Histopathological effects of cyphenothrin on the gills of Lebistes reticulatus. AB - We observed the histopathological effects of several doses of cyphenothrin. Adult guppies (Lebistes reticulatus) were placed in water containing 34.8, 46.5, 53.5 or 60 micrograms cyphenothrin/L for 96-h and their gills examined. The most common changes at all cyphenothrin doses were the lifting of the epithelial layer from gill lamellae and some necrosis. Degeneration of secondary lamellae due to edema, the shortening of secondary lamellae, and club-shaped lamellae were also observed. The 96-h LC50 for cyphenothrin to Lebistes reticulatus was 48.7 micrograms/L. PMID- 10670077 TI - Black walnut induced laminitis. AB - A 5-y-old Paint horse gelding was evaluated for acute laminitis after exposure to black walnut shavings. The gelding's feet were previously soaked in an ice bath continuously for approximately 24 h. Treatment consisted of anti-inflammatory and vasodilator therapy. Serial radiographs revealed progressive palmar deviation of the third phalanx and subsolar abscesses in both forefeet. The gelding developed purulent discharge from the right coronary band and the hoof wall detached circumfrentially. Euthanasia was elected after 54 days. Continual exposure of the gelding's feet to ice water temperatures may have caused decreased perfusion and increased edema formation in the laminae resulting in decreased blood flow and exacerbating the existing ischemic necrosis. PMID- 10670078 TI - Anaphylactoid reaction to rattlesnake envenomation. AB - The clinical manifestations of an anaphylactoid reaction are identical to true anaphylaxis; however, a previous exposure to the offending agent is not needed to manifest these symptoms. We present a case of an anaphylactoid reaction in a 62-y o female following a first-time envenomation by a rattlesnake. The patient required s.c. epinephrine and i.v. diphenhydramine, methylprednisolone, and ranitidine. She had not been envenomated by a rattlesnake previously or received any horse-derived antivenins in the past. PMID- 10670079 TI - Cestrum laevigatum poisoning in goats in southeastern Brazil. AB - Natural and experimental poisonings by Cestrum laevigatum are described in goats. Histologically, livers had marked centrolobular and midzonal coagulative necrosis and hemorrhage. Spontaneous toxicosis by this plant in goats has not been previously reported. PMID- 10670080 TI - Topical absorption of isopropyl alcohol induced cardiac and neurologic deficits in an adult female with intact skin. AB - Topical exposure to isopropyl alcohol has been reported in the literature to be toxic if sufficient isopropyl alcohol is absorbed (1-5). A clinical case is reported where a 48-y-old female presented with multiple unexplained cardiac and neurological deficits. The woman had developed the deficits over a 6-mo period in which she had been soaking towels with isopropyl alcohol and applying then to her skin overnight to ease arm pain she was experiencing. Cessation of the isopropyl alcohol exposure resolved her deficits within 3 d. A controlled repeat dermal exposure to isopropyl alcohol under clinical observation reproduced the deficits noted with corresponding serum and urine concentrations of isopropyl alcohol and acetone. Cessation of topical isopropyl alcohol exposure lead to subsequent resolution of all toxicities. PMID- 10670081 TI - Weakness, tremors, and depression associated with macadamia nuts in dogs. AB - The ASPCA National Animal Poison Center managed 29 cases of ingestion of commercially available macadamia nuts in dogs during a 5-y period. Clinical signs included, from most to least, weakness, depression, vomiting, ataxia, tremor, hyperthermia, abdominal pain, lameness, stiffness, recumbency, and pale mucous membranes. The onset of clinical signs was reported as < 12 h in 79% of the cases. The duration of clinical signs for the majority of cases was < 24 h. The amount of macadamia nuts ingested was estimated in 72% of the calls with a mean of 11.7 g/kg bw. In an attempt to reproduce the syndrome, 4 dogs were gavaged with 20 g macadamia nuts/kg bw in a water slurry. The experimentally dosed dogs developed weakness, manifested by the inability to rise 12 h after dosing, mild central nervous system depression, vomiting, and hyperthermia, with rectal temperatures up to 40.5 C. Mild elevations in serum triglycerides and serum alkaline phosphatase were detected. Lipase values peaked sharply at 24 h and returned to normal by 48 h after dosing. Other serum biochemical and electrolyte determinations were unremarkable. Serum lipoprotein electrophoresis determinations were unchanged from baseline. The mechanism of the syndrome is unknown. All field and experimental dogs recovered uneventfully within 1 to 2 d whether treated by a veterinarian or not. PMID- 10670082 TI - Mercury contamination of heavy metal collection containers. AB - We investigated discordant urinary mercury testing results from 2 patients with potential mercury exposures. Two patients had mercury levels of 634 and > 1,000 micrograms/L respectively. Although repeat 24 h urine mercury levels were elevated, spot urines were negative. Investigation revealed that technical HCl with high mercury content had been added to the 24 h urine collection containers. Subsequently, 20 hospitals were contacted to determine their heavy metals testing procedure and to analyze the acid used for mercury. Most hospitals contacted used acid in the preparation of their urine heavy metal collection containers. Of 13 HCl samples tested, 5 had low levels of mercury and 1 had heavy mercury content. Acid added to heavy metal collection containers should be of high purity grade to avoid mercury contamination of samples. PMID- 10670083 TI - A newly emerging toxic dinoflagellate, Pfiesteria piscicida: natural ecology and toxicosis to fish and other species. AB - Pfiesteria, a toxic dinoflagellate, recently has emerged as a cause of fish kills near the East Coast. Recent research into one species. Pfiesteria piscicida, has revealed a complex life cycle of at least 24 stages. Metamorphosis of one stage to another often depends on presence or absence of fish. Growth of P piscicida is promoted both directly and indirectly by nutrients such as inorganic phosphate and nitrate, as well as organic phosphate, and may be related to effluent-induced blooms. Sewage and agricultural runoff flowing into estuaries often provide these nutrients and may be correlated with the majority of fish kills in the Atlantic coastal region of the US (5). P piscicida is extremely toxic, with a low density capable of killing fish within 3 minutes (1,3,12). Fish exposed to sublethal doses of the toxin have prominent lesions. The syndrome leads to population level death losses and associated economic losses in local fisheries. PMID- 10670084 TI - Ivermectin: an assessment of its pharmacology, microbiology and safety. AB - Ivermectin (IVM) is the drug of choice for a variety of parasitic diseases due to its broad spectrum of activity and wide margin of safety. More than 18 million people are treated with IVM each year. Delivery modes include oral, topical, and s.c. injections. Its anti-parasitic activity depends upon species and developmental stages. Although IVM is believed to act mainly through interactions with invertebrate glutamate-gated chloride (GluCl) channel, other targets such as spleen cells and aminobutyric acid receptors may play important roles in the anti parasitic activity of IVM. As several organisms have evolved resistance to IVM through mutations in p-glycoproteins and/or the GluCl channel itself, research continues on improvement of IVM either through mode of administration or the feasibility of alternative macrolides. An understanding of IVM's pharmacology is essential before improved therapeutics are created. PMID- 10670085 TI - Finding an optimal dose: considerations in accurate opioid dispensing. AB - Morphine sulfate is widely prescribed for the relief of moderate to severe acute and chronic pain. Unfortunately, medication errors associated with morphine sulfate can result in patient harm; profound respiratory depression can result from excessive doses in opioid-naive patients. An actual medication order containing a potentially toxic overdose of morphine sulfate is examined. Patients habituated to high doses of morphine sulfate are at risk from underdosing, a hazard also of concern. PMID- 10670086 TI - Examining the contribution of infant walkers to childhood poisoning. AB - Parents frequently utilize baby walkers in their infants of approximately 5-15 mo of age and create opportunities for traumatic accidents. Healthcare professionals have tried to increase awareness of their dangers; despite this, between 1986 and 1991 reported walker-related accidents rose 45%. We determined if walkers were a significant contributor to childhood poisonings and what toxins were encountered most commonly. A 14-mo prospective study in a regional poison information center determined the prevalence of accidental pediatric poisonings in children aged 5 15 mo old who suffered their exposure while in a baby walker. The regional poison information center managed 7.058 poisoning exposures, 2.8% of which occurred while the child was in an infant walker. The mean age was 8.25 mo (range 5-14 mo), with 96% less than 12 mo. Substances involved were: plants 56.7%, cleaning products 9.9%, cosmetics 5.5%, construction supplies 5.0%, cigarettes 4.5%, topicals 4.5%, oral medications 2.0%, chalk 2.0% and miscellaneous 9.9%. The majority (95%) of children were asymptomatic. Infant walkers contributed substantially less to infant poisonings than was anticipated. Despite the innocuous nature of exposures, a vulnerable population was exposed to potential poisons within reach of their grasp. Baby walker injuries are not limited to trauma, and accidental poisonings should be included in the admonitions that accompany their use. PMID- 10670087 TI - Lead contamination of imported candy wrappers. AB - Lead toxicity in a young Hispanic woman from sucking on a terra cotta candy container led to investigating lead contamination in candy packaging materials imported from Mexico. Printed cellophane candy wrappers may present a significant risk for lead exposure. PMID- 10670088 TI - Characterization of US poison centers: a 1998 survey conducted by the American Association of Poison Control Centers. AB - A 1998 survey of all 73 US poison centers, including 52 certified centers and 21 noncertified centers, is presented. Despite a continued decline of the number of poison centers operating in the US, the volume of calls has steadily increased. In 1997 these centers handled 3.65 million telephone consultations, including 2,475,010 human poison exposure cases, 134,646 animal poison exposures, and 1,036,148 information calls. Nearly the entire US population had access to a poison center (99.9%), although only 78.5% of the US population was served by a certified center. Certified poison centers handled 83.6% of human poison exposure cases reported to US poison centers. Calls to certified centers were twice as likely to be handled by staff who were certified as specialists in poison information. On average, poison center utilization was 9.2 human exposure consultations/1,000 population. Total national poison center expenses approached $81 million. The average cost/human exposure case was $33.30 in certified centers, a substantial savings when compared with the alternative of emergency department management. State governments provided the single largest source of funding. Poison center funding remains unstable, with 41% of centers reporting a possible or definite budget reduction anticipated in the next budget year. In the past 5 y, 47.9% of centers faced threat of closure. Center certification and increased public education activity, especially the distribution of poison prevention materials and number of media contacts, were associated with greater utilization of the poison center in the region served. PMID- 10670089 TI - A very long trip to gastric decontamination! PMID- 10670090 TI - [Surgical infections. When is a minimally invasive procedure indicated?]. PMID- 10670091 TI - [Multiorgan surgery in rectal cancer--extended therapy or improvement of prognosis?]. AB - About 10% of rectal cancers have to be treated by a multivisceral resection. Within a period of 14 years we performed 103 of these operations, 60 for primary cancers, 43 for recurrent cancers with a significant increase in the latter. The total or partial removal of 204 extra-rectal organs allowed for a R0 resection in 67% of cases. 69% were confirmed as being pT4 by histologic examination. In comparison with conventional rectal resection morbidity is higher at 32% while mortality is identical with 4%. A significant benefit (p < 0.05) in terms of survival is found for primary cancers as opposed to recurrences, R0 resections compared to R1 or R2 resections, lymphnode negative compared to lymphnode positive patients and patients who underwent IORT. The number of additionally removed organs did not influence survival. Compared with the natural course even palliative resections carry a survival benefit and allow the treatment of the often devastating specific morbidity of the disease. Despite adjuvant multimodal therapies the high rate of local and distant recurrences after multivisceral resection of the rectum still poses a major problem. PMID- 10670092 TI - [Tumor genesis and prognostic factors in colorectal carcinoma with special consideration of tumor localization]. AB - The rising incidence of colorectal carcinoma, particularly in the industrial nations of Europe and USA, directs the attention to the aetiological factors of these tumors: nutrition, the association with colorectal adenoma, familiarly genetic disorders as familiar adenomatous polyposis and hereditary non polyposis colorectal cancer. Some aspects of molecular biology are discussed. Furthermore, right and left part of the colon (divided by the Cannon-Boehm point) and the neoplasms of these sections of the colon are different in embryology, function and morphology. The cancers of the right colon develop without polypoid changes, those of the left part in majority via the adenoma-carcinoma sequence. It is possible to demonstrate differences between these two localizations in the DNA content (diploid tumors on the right side), in the lost of allels (especially distal tumors), in proliferation activity (lower in right side tumors) and in the expression of oncofetal antigens. Besides, there are some histological differences between neoplasms of the right and left colon (production of mucin, "Crohns like lymphoid reaction" histological grading). In our own material of 262 patients with resected colon carcinoma we have investigated the distribution of carcinoma in the right and left colon, furthermore the T-classification, histological grading and the proportion of mucinous carcinoma in the different tumor localizations. PMID- 10670093 TI - [Surgical management of locoregional recurrence of gastric carcinoma]. AB - About half of the patients with gastric cancer subjected to total gastrectomy in curative intention die of recurrence within a few years. Most of these local recurrences occur in the first 2 years postoperatively. In an historic analysis 133 patients of the years 1985-1997 were investigated. Local recurrence was observed in 29 cases within 60 months on average. An intensive follow-up will not affect the long-term outcome of local recurrence. Improved results may be expected only if more effective therapeutic strategies for local recurrence will be developed. PMID- 10670094 TI - [Demand for and use of blood supply for elective surgical procedures]. AB - In elective surgical operations on thyroid gland and breast gland, in cholecystectomy, axillary or inguinal dissections and hernioplasties blood units are usually ordered for the operation. The aim of the study was to analyse the real requirement of transfusions during several years and to show that in the above-mentioned operations only in exceptional cases blood units must stand by. METHODS: At the Surgical Hospital I of the University Leipzig, a retrospective analysis of the anaesthetic records and patient documentations from 1994 to 1997 was performed with regard to intraoperative blood transfusions. RESULTS: There were 1122 operations on the thyroid gland (119 of it as total thyroidectomy), 465 operations on the breast gland, 413 cholecystectomies, 70 axillary and 60 inguinal dissections and 445 hernioplasties. Intraoperative transfusions were necessary in nine operations on the thyroid gland (0.8%), in six operations on the breast gland 1.3%), twice in cholecystectomy (0.5%) and only once in an axillary dissection (1.4%). The analysis of the patients' records showed in almost all of these cases special risk factors such as disorders of blood coagulation or thyroidal function, anaemia, serious other diseases or a necessary extension of the operation. CONCLUSION: It is justified to perform the above mentioned operations without a routine order of blood units. This would lead to enormous financial savings. Because the optimal care for the patients has priority, it is necessary to estimate the individual risk of a required transfusion preoperatively and to keep low the blood loss by the surgeon during the operation. PMID- 10670095 TI - [Is there a chronic appendicitis in childhood? Analysis of pediatric surgical patients from 1993-1997]. AB - The right lower quadrant abdominal pain and the "chronic" appendicitis in childhood and adolescence are frequently connected with a lot of different diagnostical problems for the treating physician. Since the introduction of diagnostical laparoscopy and laparoscopical appendectomy in our hospital the rate of appendectomy has been increased to 35% in case of the histological diagnosis of "chronic" appendicitis. A special problem in childhood and adolescence is the request of the parents for clarification of chronic recurrent pain in their children and therefore the demand of laparoscopy and not seldom appendectomy. After appendectomy with the pathological-histological findings of "chronic" appendicitis children are in 83% without any abdominal pain. Therefore the question arises whether there exists a "chronic" appendicitis in childhood justifying surgery in these cases. Although the rate of complications and conversions (3.8%) in diagnostical laparoscopy and laparoscopical appendectomy is quite low the indication should only be made after careful examination considering differential diagnostical problems. In our hospital 56.5% of the children with the histological diagnosis of "chronic" appendicitis suffered retrospectively on other diseases, that had been caused the symptoms of appendicitis. Before operation the patients should be informed on the complications in detail. PMID- 10670096 TI - [Chronic appendicitis. Recurrent abdominal pain in the right lower quadrant from the viewpoint of the internist]. AB - There is no clear scientific evidence for a clinically relevant chronic form of appendicitis in the absence of acute flares. Lacking typical symptoms of acute appendicitis or corresponding imaging findings, no indication is given for appendectomy from the internal medicine point of view. By contrast, chronic or recurrent right lower quadrant pain is often of functional origin and may be part of the Irritable Bowel Syndrome or the Functional Abdominal Pain Syndrome. These syndromes are linked to a higher rate of appendectomies in the medical history. The Irritable Bowel Syndrome may be diagnosed based on clinical symptoms alone. But in doubt and in considering malignancy, the indication for diagnostic imaging is given, after ultrasound particularly by colonoscopy. For positively diagnosing these functional syndromes, the typical clinical presentation, extraintestinal pain syndromes, and psychic factors should be evaluated. The visceral hypersensitivity is the predominant pathophysiologic finding and measured by rectal distention stimuli. Medical treatment comprises relaxatives of smooth muscle and low dose antidepressants as modulators of visceral perception. These are supplemented by the psychosocial management. PMID- 10670097 TI - [Differential diagnostic problems of hepatocellular carcinoma in an abnormal additional liver lobe in the lower abdomen]. AB - This case report describes a 36 years old patient with a known tumor in her lower abdominal cavity which increased in size in the last 16 years. Intraoperatively an additional liver lobe of the left liver located in the lower abdominal cavity was found in combination with a multifocal hepatocellular carcinoma which could be treated curatively. PMID- 10670098 TI - [Localized Castleman disease. Diagnostic difficulties in a surgically treatable disease]. AB - Castleman's disease is a rare cause for a retroperitoneal mass. The disorder can be classified into two histopathological groups: the hyalin-vascular and plasma cell types. The former type, particularly in its localized form, is often asymptomatic and benign, the surgical therapy may cure the patient. The latter type is frequently associated with systemic manifestations and an uncertain prognosis. We present a case in which the diagnosis of localized Castleman's disease proved to be extremely difficult. In these cases, the correct diagnosis, however, is indispensable, as these patients can be cured by radical surgical tumor excision. PMID- 10670099 TI - [Thoracoscopic treatment of pleural empyema]. AB - From August 1991 to May 1997 46 patients with pleural empyema in the fibrinopurulent phase underwent thoracoscopic surgery. There were 36 men and 10 women with an average age of 47 years ranging from 18 to 84. The average operating time was 77 minutes. When only one thoracostomy drain was inserted, the drainage time was 8.5 days, if two or three drainage tubes were used it was 10.5 days. The average hospital stay was 18.1 day (range from 7 to 45). We observed ten complications. Four operations had to be converted to an open procedure because of massive thickening and fibrosis of the pleura. Three patients did not tolerate one lung ventilation, once the lung did not collapse due to technical reasons and in one patient each we observed a laceration of the parenchyma and bleeding from the parenchyma. In both cases the problem was dealt with thoracoscopically. We observed a recurrent pleural empyema in four patients which occurred between the 28th and 77th postoperative day. In summary, thoracoscopic surgery in patients with pleural empyema in the fibrinopurulent phase is an effective and well tolerated alternative to open thoracotomy. PMID- 10670100 TI - [Laparoscopic cholecystectomy. A recommendable indication in acute cholecystitis?]. AB - While laparoscopic cholecystectomy has become the procedure of choice for the elective treatment of symptomatic cholecystolithiasis the question whether patients with acute cholecystitis should be operated laparoscopically or conventionally is still debated. Nevertheless, more and more surgeons tend to use the laparoscopic approach even in patients with acute cholecystitis. Of 1006 laparoscopic cholecystectomies performed at our hospital 42 were done for acute cholecystitis. Conversion to an open procedure was necessary in only one patient because of severe inflammatory changes. The overall mortality was zero. The average age was 45.9 years for all patients and 50.4 years for those with acute cholecystitis. The average operating time in patients with acute cholecystitis was 81 minutes compared to 62 minutes in patients who underwent elective laparoscopic cholecystectomy. The complication rate and the average hospitalization time did not differ significantly between the two groups. Our own data as well as the data retrieved from the literature seem to indicate that laparoscopic cholecystectomy is superior to the open procedure in the treatment of acute cholecystitis. Prerequisite is that the operation is performed less than 72 hours after the onset of the symptoms by an experienced operating team and the readiness to convert to open procedure if necessary. Under those circumstances laparoscopic cholecystectomy seems to be the treatment of choice for acute cholecystitis. PMID- 10670101 TI - [Role of laparoscopy in the management of acute appendicitis]. AB - Minimal invasive surgery had a considerable impact on common surgical techniques and has almost replaced established operative procedures such as cholecystectomy. However, the laparoscopic approach for the treatment of acute appendicitis is still not very popular. We discuss the role of laparoscopy for appendectomy and include three studies from our institution (University Hospital Zurich, Switzerland) and prospective studies reported in the literature. We conclude that laparoscopic appendectomy, when compared with the open approach, has the following advantages for the diagnosis and treatment of acute appendicitis. (1) Diagnostic laparoscopy is an effective and relatively atraumatic tool to investigate the abdominal cavity, which results in a sensitivity of almost 100%. This allows for accurate decision making, which is especially advantageous in young women and obese patients. (2) Prospective studies demonstrate that laparoscopic appendectomy is at least as good as open appendectomy and that the laparoscopic approach results in a reduced postoperative infection rate. (3) The similar complication rate after laparoscopic appendectomy, when performed by residents rather than staff surgeons, underlines the feasibility and teaching potential of this minimal invasive procedure. PMID- 10670102 TI - [Laparoscopy in intra-abdominal infection. Its diagnostic value and therapeutic possibilities]. AB - In 550 patients with the clinical features of acute abdomen a surgical laparoscopy was performed. In 121 cases there was found an unspecific reason of the acute abdominal disease that did not require surgical therapy. In 349 cases a regional peritonitis was found, 80 times a diffuse peritonitis. The diagnostic validity of laparoscopy was 96% as compared to 42% for sonography. The laparoscopic access resulted in a complication rate of 0.2%. In 239 cases (43%) the disease could be managed laparoscopically, 190 cases (35%) required open surgery. PMID- 10670103 TI - [Laparoscopic management of echinococcal cysts of the liver]. AB - Even though echinococcal cysts have been cured by puncture and instillation of scolicidal medications, surgery is still the mainstay of the treatment of hydatid disease. The aim of the surgical treatment is the elimination of scolices, the removal of all viable parts of the cyst and the obliteration of the remaining cavity. This can be achieved by resective procedures, but also by a more conservative approach with drainage and obliteration of the cyst. The latter procedure can be done by open surgery or laparoscopically. The disadvantages of the laparoscopic approach are the increased danger of contamination of the abdominal cavity with scolices and difficulties to aspirate a highly viscous cyst content. Furthermore, cysts which are located deep in the parenchyma of the liver should not be approached laparoscopically because of the significant danger of hemorrhage. The advantage of the laparoscopic approach in selected cysts, i.e. those which are located superficially and having a liquid content, are a shorter hospital stay, lower incidence of wound infection and the ability of the surgeon to inspect the inside of the cyst more thoroughly and rule out daughter cysts and connections to the biliary tract. A review of the literature (n = 76) indicates that in most laparoscopically treated hydatid cysts of the liver a simple drainage (59%) or an unroofing (31%) is performed. The complication rate is 21%. Because there are no longterm observations after laparoscopic operations for hydatid disease the question of recurrence cannot yet be answered. PMID- 10670104 TI - [Laparoscopic sigmoid resection in diverticulitis]. AB - It is still difficult to determine the exact indication for a laparoscopic sigmoid resection for diverticular disease. Frequently, the severity of diverticulitis is not sufficiently defined. For this reason a modification of the Hinchey classification is proposed to which a stage II b for fistula formation and a differentiation between acute and chronic disease have been added. Another problem is the lack of criteria which define a "laparoscopic" resection. A sigmoid resection should be called "laparoscopic" if the mobilization of the sigmoid colon, the transsection of the mesenteric vein and artery and the mesentery itself and the distal transsection of the bowel are done laparoscopically. The resection of the bowel and the introduction of the anvil of the stapler device can be done extraabdominally, however, the anastomosis again should be performed laparoscopi-cally. A so defined sigmoid resection can be done in the chronic stage I. In the chronic stage II a there will be significant problems due to adhesion formation, and in the acute stages II a and II b as well as in the chronic stage II b a laparoscopic resection should not be attempted. PMID- 10670105 TI - ["Identification of of the recurrent laryngeal nerve through intraoperative neuro monitoring."]. PMID- 10670106 TI - ["Medical malpractice in laparoscopic cholecystectomy."]. PMID- 10670107 TI - [Therapeutic principles in extensive soft tissue injuries and basic principles of plastic defect covering]. PMID- 10670108 TI - [General infection prevention in abdominal surgery with special reference to intestinal decontamination]. AB - In surgery prophylaxis for infection is necessary, because patients are immunocompromised due to the underlying disease and the operation while at the same time being increasingly exposed to potentially pathogenic germs. Prophylaxis is based on the control of endogenous and exogenous microorganisms. For this purpose either systemic or locally active topical agents may be employed. Systemically active substances are applied with the aim to kill and eliminate invasive microorganisms in deep tissue levels, either by their own biological activity or by stimulating specific or unspecific host immune reactions. Local topical measures in contrast are to prevent the primary contact between microorganisms and host. The central pillar of systemic measures is the perioperative systemic antibiotic prophylaxis, immunonutrition is beginning to gain importance, and in the future possibly substances such as G-CSF, which directly stimulate the immune system, may be employed. Standard topical measures are sterilization and desinfection while decontamination of the digestive tract has until now not found a wide spread acceptance. For certain indications especially high risk surgical resections with anastomoses at the level of the oesophagus or the lower rectum it is possible to eliminate endogenous intestinal microorganisms effectively using topical decontamination in combination with systemic antibiotics and improve the surgical results, especially anastomotic healing. PMID- 10670109 TI - [Perioperative infection prophylaxis in gastrointestinal surgery]. AB - Antibiotic perioperative prophylaxis is known to reduce postoperative infections and is generally administered as single-dose regimen today. Effective prophylaxis requires plasma- and tissue concentrations above the MIC of the expected bacterial spectrum throughout the whole operation from skin incision to wound closure. In longlasting operations a second dose after 4 hours is recommended. Indications for antibiotic prophylaxis are clean-contaminated or contaminated procedures according to Cruse and in patients with elevated risk of infection. Generally antibiotic prophylaxis is administered intravenously. Chinolons achieving high tissue- and plasmalevels and showing a broad antibacterial spectrum seem appropriate to be used for oral prophylaxis. In a prospective study in 36 patients pharmocokinetics of ofloxacin were measured perioperatively and proper plasma- and tissue concentrations were attained. A following prospective randomized study in 56 patients undergoing colonic or pancreatic resections oral prophylaxis did not show a higher infection rate compared with a standard intravenous prophylaxis. Therefore oral prophylaxis seems to be an attractive option, its effectivity needs to be proven by studies with sufficiently high patient numbers. PMID- 10670110 TI - [The influence of taurolidine on physiological and pathological blood coagulation and implications for its use]. AB - Taurolidine is an anti-infective agent with an unusually broad spectrum of effectivity against gram-positive and gram-negative bacteria, anaerobic organisms, and fungi. The effective principle is explained by the decomposition of the substance and the transfer of methylol groups to specific molecular structures of the cell walls of microbes. The acceptors are amino groups of the amino sugars and the lysine residues of glycoproteids. More recent investigations have shown that the effect of taurolidine is not limited to microorganisms, but can be detected in cells of the macrorganism as well. Here the influence of taurolidine on different blood clotting factors is described. The results can be explained by a transfer of methylol groups to residues of arginine and histidine in the active region, in analogy to the transfer to lysine residues in microorganisms. It is therefore to be expected that taurolidine will influence other vital systems of the macroorganism, dependently of concentration, as long as their biological function is connected to residues of arginine or histidine. Examples are the complement system and the fibrinolysis system. The implications of these observations have to do with new indications in connection with clotting phenomena in extracorporal circulatory systems or with thrombolysis, as well as with known indications in cases of shock and sepsis. PMID- 10670111 TI - [Treatment of septic complications od secondary peritonitis]. AB - The authors review the current management goals of surgical and antibiotic therapy in secondary peritonitis. Basic therapeutic regimen is the surgical elimination of the infectious source by means of a rational and risk-adapted operative procedure. Other technical procedures (such as intra- and postoperative lavage, drainage of the abdominal cavity, decompression, etc.) are critically reviewed and reduced to the scientific background. A calculated antibiotic therapy should be performed to eliminate the leading pathogens. The golden standard seems to be the short-term application of a third generation cephalosporin in combination with metronidazole. Antibiotic-induced endotoxinemia, bacterial shifting to grampositive species, and escalating antibiotic resistance require the examination of new therapeutic regimens (e.g. with quinolones). PMID- 10670112 TI - [Prophylaxis and therapy of infectious complications of lung surgery]. AB - Postoperative infections are a dreaded complication in pulmonal surgery. Besides the optimal preparation of the patients and careful operative technique, perioperative antibiotic prophylaxis represents an important factor in avoiding infectious consequences. Owing particularly to the high proportion of patients with malignant, consumptious illnesses in thorax surgery, immune deficiencies must be reckoned with in this group of patients. The spectrum of germs to be expected within the framework of pulmonal surgery determines to some extent which antibiotic shall be used. We have investigated the efficacy of a standardized antibiotic prophylaxis using cefotaxime (Claforan) in 200 pulmonal patients. Pleural empyema is a rare, but nonetheless important infectious illness, as a consequence of pulmonal operations, or also following pneumonia. Whilst the early stages of an empyema can often be successfully treated using only drainage treatment, chronic empyema usually requires a thoracotomy with empyema dissection and excortication, as well as subsequent irrigation-suction drainage treatment. In spite of specific surgical sanitation and irrigation-suction drainage treatment, therapy is often complicated by persistent germs in the thoracic cavity. Instillation therapy with taurolidine can lead to faster healing of the infection in such cases. Purulent mediastinitis is an extremely rare illness, but dreaded owing to its high mortality. The causes of the illness lie in injuries of the trachea, of the bronchial tubes, and of the oesophagus. With the introduction of medial sternotomy as operative entry, mediastinitis as a postoperative complication has increased noticeably in frequency. Mediastinitis occurs as a descending infection as a consequence of odontogenic affections. Owing to frequently late diagnosis, infection is usually advanced, so that simple drainage treatment of the mediastinum no longer suffices in many cases. We introduce our concept of treatment using our own patient collective. PMID- 10670113 TI - The role of NMDA antagonist in perioperative pain management. PMID- 10670114 TI - Comparison of rocuronium and vecuronium pretreatment for prevention of fasciculations, myalgia and biochemical changes following succinylcholine administration. AB - BACKGROUND: The purpose of this study was to assess the effect of rocuronium pretreatment on the succinylcholine-induced fasciculations, myalgia and biochemical changes, and to compare it with vecuronium pretreatment. METHODS: We have studied 60 female patients undergoing minor elective surgery, in a prospective double blinded method. Three groups of 20 patients each was pretreated with either saline (control group), rocuronium 0.05 mg/kg (rocuronium group) or vecuronium 0.007 mg/kg (vecuronium group). Three min after the pretreatment, 1.5 mg/kg succinylcholine was injected. Single twitch responses to electrical stimulation were measured. Serum potassium and creatine kinase were respectively measured 5 min after succinylcholine and 24 h after operation. Fasciculations and myalgia on postoperative day 1 and day 2 were evaluated. RESULTS: The incidence of fasciculations was lowest in the rocuronium group, followed by the vecuronium group, and was highest in the control group. The incidence of myalgia on postoperative day 1 was lower in the rocuronium and vecuronium groups than the control group. The increase of serum creatine kinase was similar among the three groups, but there was no increase in serum potassium concentration in any group. No differences of the single twitch responses to electrical stimulation were found between the rocuronium and vecuronium groups. CONCLUSIONS: Rocuronium pretreatment was more effective in reducing fasciculations than was vecuronium pretreatment, but both were equally effective in preventing myalgia on postoperative day 1. This difference may reflect the differential activities of rocuronium and vecuronium at the neuromuscular junction. The increase of creatine kinase was not attenuated by any regimen. PMID- 10670115 TI - Postoperative intramuscular dextromethorphan injection provides postoperative pain relief and decreases opioid requirement after hemorrhoidectomy. AB - BACKGROUND: Previous studies have shown that dextromethorphan (DM) produces an analgesic/antihyperalgesic effect. This study was designed to examine whether postoperative DM intramuscular (i.m.) injection could reduce post hemorrhoidectomy pain. METHODS: At the end of the surgery, patients in the study group (n = 30) were given an intramuscular injection of 40 mg DM and 20 mg chlorpheniramine (CPM) while in the study group (n = 30), the patients were given intramuscular 20 mg CPM only. Pethidine (1 mg/kg, i.m.) was prescribed for postoperative pain relief if required. The time to first pethidine injection, total pethidine consumption, worst pain score, and pethidine-related side effects were recorded for 48 h postoperatively. RESULTS: The time from the end of operation to the first pethidine injection was 5.4 +/- 1.6 h and 17.8 +/- 3.7 h (P = 0.006) in the control group and the study group, respectively. Total pethidine consumption was 139.5 +/- 11.5 mg and 77.5 +/- 12.2 mg (P < 0.001) in the control group and the study group, respectively. The worst VAS score was 7.5 +/- 0.2 and 7.1 +/- 0.2 (P = 0.09) in the control and the study groups, respectively. The number of patients who required pethidine injection was 29 and 21 (P < 0.005) in the control and the study groups, respectively. The number of patients who suffered pethidine-related side effects was 7 and 1 (P < 0.025) in the control and the study groups, respectively. CONCLUSIONS: We found that intramuscular DM given at the end of operation could provide good postoperative pain relief and decrease the pethidine requirement after hemorrhoidectomy. PMID- 10670116 TI - Is sedation necessary for demented patients under infiltration anesthesia? AB - BACKGROUND: Sedation for intraoperative patients under infiltration anesthesia is often used, however, disadvantages of sedation for demented patients may sometimes exceed its advantages because some of the demented patients are already apathetic about their surroundings. The authors prospectively investigated whether sedation for alert or demented patients receiving surgery under infiltration anesthesia is useful and safe for intraoperative management. METHODS: Sixty patients undergoing irrigation and drainage of chronic subdural hematoma under infiltration anesthesia were divided into four groups. Patients in Group A were non-sedated alert patients. Patients in Group B were non-sedated demented patients. Patients in Group C were sedated alert patients. Patients in Group D were sedated demented patients. Intraoperative variables, adverse effects, and the postoperative satisfaction represented by five-point score were recorded. RESULTS: The heart rate in group A during operation was significantly faster than the control value. The postoperative satisfaction score in group A was significantly lower than any other groups. A patient in group D required a nasal airway to improve respiration during operation. CONCLUSIONS: These results suggest that there is no evidence to show that advantages of sedation for demented surgical patients under infiltration anesthesia excel the disadvantages although sedation for alert patients may be an effective and rational conduct in intraoperative management. PMID- 10670117 TI - The effects of tramadol versus fentanyl in attenuating hemodynamic response following tracheal intubation. AB - BACKGROUND: Tramadol is a novel central acting analgesic. It has been used as a complement to general anesthesia and an effective agent for postoperative analgesia. However, the influence of tramadol on the hemodynamic response following laryngoscopy and tracheal intubation is less known. METHODS: Forty patients of both sexes, 16-50 year old, ASA physical status I or II, scheduled for elective surgery were randomly divided into equal groups in this prospective, double blind study. After obtaining the baseline data, the patient was given 3 micrograms/kg fentanyl (Group F) or 3 mg/kg tramadol (Group T). Then induction of anesthesia in a uniform and standardized manner was carried out by an anesthesiologist who was blind to the medication. The hemodynamic parameters were measured and recorded immediately after induction but prior to laryngoscopy, 3, 6, and 9 min after intubation, and before incision. We also observed any unusual effect in the postoperative care unit. Chi-square test, Student's t-test and paired t-test were used for statistical comparison. A P less than 0.05 was considered statistically significant. RESULTS: All patients had a successful induction and intubation. Differences in baseline values were not significant, nor were the differences in the values following induction. After laryngoscopy and intubation, heart rate increased significantly above the baseline level in both groups. The increase of heart rate was significantly more at 6 and 9 min (P < 0.05) and lasted longer in the tramadol group. After intubation, systolic, mean and diastolic arterial pressure (SAP, MAP, DAP) increased significantly above baseline in both groups too, except for DAP in fentanyl group. At 6 and 9 min, the MAP and DAP were significantly higher in tramadol than in fentanyl group (P < 0.05). Six patients in tramadol group had mild pain on injection of tramadol. CONCLUSIONS: When administered right before thiopental induction, 3 mg/kg tramadol did not display a better attenuation against the increase of hemodynamic profiles than did 3 micrograms/kg fentanyl following tracheal intubation. PMID- 10670118 TI - New frontiers in mechanisms and therapy of painful peripheral neuropathies. AB - Ten years ago we had no more than informed speculations about the pathogenesis of neuropathic pain, and our ability to give these patients relief was so limited that neuroablative therapy or even deep brain stimulation via an implanted electrode was often the only hope. We now have a thorough emperically based, although undoubtedly still incomplete, understanding of the pathophysiology. Moreover, we have discovered new classes of drugs that specifically relieve neuropathic pain. It is not unreasonable to hope that in the near future patients with painful peripheral neuropathy will never again have to suffer unrelieved pain. PMID- 10670119 TI - Acute fatal vasoplegia and asystole induced by intravenous amiodarone after cardiopulmonary bypass in a patient with preoperative cardiogenic shock. AB - Single dose intravenous amiodarone has been widely used and shown to be effective to treat supraventricular and ventricular arrhythemias in cardiac surgery. We, herein, report a 60-year-old female patient, sustaining cardiogenic shock in the course of percutaneous transluminal coronary angioplasty (PTCA) for unstable angina unrelieved by medication including nitroglycerin, succumbed to a life saving emergent coronary artery bypass grafting (CABG) operation at the end of cardiopulmonary bypass (CPB) following a 180 mg bolus dose of amiodarone (3 mg/kg) directed at the ventricular arrhythmias, triggered by protamine and unresponsive to lidocaine treatment. Amiodaroneinduced asystole and vasoplegia were thought to be the causation of the failure of resuscitation. The causes of the development of these complications, the potential hazards of its use and the management relative to the consequential complications are reviewed and discussed. PMID- 10670120 TI - Postoperative myocardial infarction in a patient with perioperative ST-depression -a case report. AB - Often we ignore electrocardiogram (EKG) evidence of ischemia and no adverse events occur. However, once in a while these ischemic episodes will turn into a full-blown myocardial infarction. Therefore, studying perioperative events which tilt the balance over to postoperative myocardial infarction (PMI) can enlighten our knowledge in postoperative myocardial infarction (MI) prevention. We present a case of ST depression in perioperative EKG evolving into postoperative MI. In this paper we attempt to explore various possibilities which could have altered this patient from her ischemic state into an infracted event. PMID- 10670121 TI - Postoperative right atrial and pulmonary embolism after prolonged spinal surgery. AB - Perioperative pulmonary thromboembolism can proceed rapidly with grave prognosis, in which immediate or accurate diagnosis and management is not easy. According to the literatures, patients receiving spinal surgery are at relatively lower risk of developing thromboembolism. We would like to present a case of postoperative pulmonary thromboembolism which developed after a prolonged lumbar spinal surgery. Tachycardia and unstable hemodynamics were noted postoperatively. Pulmonary and right atrial thrombi were disclosed by transesophageal echocardiography. Although cardiotomy and thrombectomy were immediately performed, the patient finally died 3 days after the operation. The pathogenesis of venous thromboembolism (VTE) in the surgical patients, the risk factors which predispose a patient to VTE, diagnosis, and treatment as well as the prophylactic measures of VTE are herein reviewed and discussed. PMID- 10670122 TI - Unilateral vocal cord paralysis following endotracheal intubation--a case report. AB - A 41-year-old man of ASA physical status class I was scheduled to receive the video-assisted thoracoscopic T2 sympathectomy for hyperhidrosis palmaris. The elective surgery was performed smoothly under general anesthesia with endotracheal intubation. However, the patient complained of hoarseness in the postoperative period. A stroboscopic examination showed that the left vocal cord remained stationary in the paramedian position, signifying left vocal cord paralysis. In the case, we believed it was most likely that endotracheal intubation might be responsible for the unilateral vocal cord paralysis. The possible cause was that during placement or thereafter during positioning, the endotracheal tube was malposed or slipped upward, rendering its inflated cuff to rest against the vocal cords. Another reason was that the cuff which was over inflated made the vocal cords under constant pressure. Both conditions may cause damage to the anterior branch of the recurrent laryngeal nerve. We also discussed the general management and prophylaxis for the unilateral vocal cord paralysis. PMID- 10670123 TI - [Immunohistochemistry applied to urology]. PMID- 10670124 TI - [Radical prostatectomy in clinically localized prostatic adenocarcinoma. Study of patients with positive margins and their impact on survival free from biochemical progression]. AB - OBJECTIVES: Margins involvement in T1-T2 patients undergoing radical prostatectomy is a negative prognostic factor. We aimed to: a) Study the clinical and pathological features of patients with surgical margins involvement; b) Elucidate the influence of margins involvement on the progression-free survival. MATERIAL AND METHOD: The study included the group with "positive margins" out of a series of 160 patients with localised prostate adenocarcinoma who underwent radical prostatectomy at the Clinica Universitaria de Navarra between 1988-1997. statistics used: Fisher's or Pearson's test for qualitative variables. Kaplan Meyer, Log-rank and Cox's multivariate tests for the survival study. RESULTS: The group accounts for 28% (45/158) of all patients undergoing radical prostatectomy. Mean PSA (22 +/- 14 ng/ml) is similar to the remaining group although there is greater significant rates of PSA > 15 ng/ml (p: 0.006), worse Gleason (p: 0.01), higher proportion of T2bc (p: 0.003) and node involvement (0.001). Progression free survival (BPFS) is significantly lower in this group (32 +/- 12% vs 61 +/- 6% at 5 years). Margins are the single factor with higher influence (RR:5) in the multivariate study. Influence is clear in patients with Gleason < 5 (0% vs 87%) and PSA < 30 ng/ml (33 +/- 14 vs 70 +/- 7%), but has no influence on BPFS of patients with PSA > 30 ng/ml or Gleason 5-10. CONCLUSIONS: Positive margins in patients undergoing radical prostatectomy is associated to higher PSA, worse Gleason and higher stage. They are the most significant independent risk factor (except for PSA > 30 ng/ml) for biochemical progression-free survival as evidenced in the multivariate study, although it is likely this influence is diluted in patients with PSA > 30 ng/ml and/or Gleason 5-10. PMID- 10670125 TI - [Randomized prospective study comparing BCG and BCG plus oral tegafur in the prevention of stage T1 superficial tumors of the bladder. Results after 2.5 years]. AB - We present our experience in eighty patients with superficial bladder cancer stage T1. They have been randomized to receive BCG 27 mg weekly x 6 and monthly until complete one year (Group A) or the same schedule plus Tegafur 800 mg daily until complete one year. Results are similar in both groups. With a median follow up of two years and a half, 33% in Group A and 20% in Group B have had recurrence; 7.6% in group A and 3% in group B have had progression in stage. Differences are not significant in both cases. Tolerance of Tegafur is good with only 11% of secondary effects. We concluded that there are no differences in both treatments but there is a trend to better results with combinant therapy. It is necessary more patients to achieve definitive results. PMID- 10670126 TI - [Incidental prostatic cancer]. AB - OBJECTIVE: To evaluate the incidence of incidental prostate cancer and PSA ability to predict its presence. MATERIAL AND METHOD: Retrospective study of 862 patients undergoing prostate surgery between 1994 and 1997, both inclusive. Digital rectal examination provided no suspicion of neoformation. Mean age was 68 +/- 7.5 years. Mean PSA, 8.3 +/- 10 ng/ml (Hybritech). 15% patients had previously undergone at least one ultrasound-guided biopsy in the peripheral area. 55% patients underwent retropubic surgery and the remaining 45% prostate transurethral resection. Ultrasound prostate volume for both patient groups was 107 +/- 63 cc and 45 +/- 25 cc, respectively. RESULTS: Incidental cancer was found in 6% patients; 65% were T1a and 35% T1b. Mean PSA concentration in cancer patients was almost significantly (p = 0.05) higher than in patients with BPH. Patients with PSA > 10 ng/ml presented a significantly higher incidence of cancer (p = 0.02). Patients with previous prostate biopsy showed a cancer incidence rate of 12% versus 5% patients with no previous biopsy (p = 0.001). CONCLUSIONS: Incidence of incidental prostate cancer was 6%. PSA was not a good predictor of incidental cancer. Patients with PSA > 10 ng/ml, showed higher incidence of cancer. Younger patients with PSA > 10 ng/ml, and at least one negative biopsy of the peripheral area should undergo biopsy of the transitional area prior to surgery. PMID- 10670127 TI - [Bone abnormalities. Muscular dystrophy and lithiasis: lithogenic factors and therapeutic difficulties]. AB - Duchenne's muscular dystrophy with kypho-scoliosis, progressive muscle weakness and abnormal fatigue of the muscles results in an immobilisation syndrome with increased bone resorption and hypercalciuria. The accompanying chest deformity alters the respiratory capacity, causing pulmonary insufficiency, acidosis and acid urine. Dorso-lumbar kypho-scoliosis, occasionally very serious, alters the status of the upper urinary tract affecting urine transportation (stasis). Thus, hypercalciuria, urinary acidosis, stasis and infection will determine the formation of urinary lithiasis that can take place in these patients. MATERIAL AND METHODS: 15 patients with a variety of myopathies (Duchenne's disease, Myasthenia gravis,...) or serious skeletal deformities with metabolic renal lithiasis (pyelic or calyceal) were seen by our group. Other patients presented post-traumatic (paraplegia, hemiplegia,...) or poliomyelitic skeletal sequels or Pott's disease, with septic lithiasis. After evaluating all likely approaches including ESWL, the latter was chosen being the least aggressive. Conventional surgery, either percutaneous or endoscopic, foretells technical problems in terms of lithiasis approach. Both the case introducing the subject, Duchenne's muscular dystrophy, with bilateral renal lithiasis and the others are a reflection of complexity of finding the righ approach for these patients, including ESWL. RESULTS: Scoliosis was not a technical obstacle, since patients could be placed in lateral/oblique position to situate the stone in the right spot for lithotrity. Debris removal was easy, with no obstructive complications, in spite of the significant immobilisation of these patients. CONCLUSION: Immobilisation syndrome, acidosis, stasis and infection could jointly determine the lithogenesis mechanism in patients with muscle diseases or serious skeletal deformities and with renal lithiasis. ESWL has an opportunity in serious cases, where other techniques including surgery have major difficulties. PMID- 10670128 TI - [Our clinical experience with the use of ++sildenafil citrate to treat erectile dysfunction]. AB - OBJECTIVE: To evaluate the response and adverse effects to treatment with Sildenefil in those patients with an erectile dysfunction who, according to our protocol, were considered as candidates for this therapeutic option. MATERIALS AND METHODS: We reviewed the clinical histories of 180 patients seen in our service from november 1988 to February 1999 as a result of an erectile dysfunction. Those patients in whom the use of Sildenefil was not contraindicated were prescribed this product at does of 50 mg. The response to treatment was subjectively evaluated based on the option of the patient when comparing his quality of life before and after treatment. RESULTS: Out of 180 patients, 144 started treatment as indicated. Of these, 67% expressed a positive response, while 33% did not respond to treatment. Adverse effects were notice by 26% of patients, but in 97% of them they were not severe enough to withdraw the medication. CONCLUSIONS: Two-thirds of 144 patients with erectile dysfunctions of heterogeneous origin responded positively to treatment with Sildenefil. One fourth of them reported some ort of adverse effect, but almost none of them stopped the medication for this reason. PMID- 10670129 TI - [Impact of transplantectomy of the first graft on the clinical course of cadaver renal retransplantation]. AB - PURPOSE: At the present time a number of prediction factors on the outcome of second cadaver renal transplants are known, in particular the influence of recipient response to the first graft. However, we do not know if retaining or removing the non-functioning organ has any repercussion on subsequent transplants. This is the object of the present study. MATERIAL AND METHODS: A retrospective study was carried out on the clinical records of 80 patients who had undergone a second cadaver renal transplant under CyA between 1985-1995 at Hospital 12 de Octubre. Data on the characteristics and outcome of the first and second transplants were collected, plus those concerning the recipient and donor. These variables were used to construct a multivariant analysis model of graft survival. The nephrectomy of the non-functioning graft was only carried out when absolutely necessary (n = 58). RESULTS: The multivariant analysis showed that the nephrectomy of the non functioning graft did not modify the risk of recipient sensitization or the probability of developing acute or chronic rejection of the second graft. However, it reduced the risk of losing this second graft at medium or long term significantly (p = 0.009). CONCLUSIONS: These data constitute the first solid argument in favor of elective nephrectomy in candidates for a second cadaver renal transplant. Nevertheless, a more thorough study should be carried out of the immunological mechanism involved before recommending this as a general procedure. PMID- 10670130 TI - [Meningeal carcinomatosis in bladder tumor]. AB - We report a strange case of a bladder whose first metastasic manifestation, after two years of the diagnosis, was a peripheric polyneuropathia. This patient was treated with immunotherapy with BCG for superficial carcinoma of the bladder during one year. Gradually central neurological symptoms appeared and the patient died one month later. A meningeal carcinomatosis was identify as the cause. No bone metastases existed, which is the most frequent way of tumours extension towards leptomeninges. We argue about the way to arrive at meninges. PMID- 10670131 TI - [Report of a new case of small-cell carcinoma of the bladder and review of the literature]. AB - We report a case of small cell carcinoma of the urinary bladder in 71-year-old male patient. Oat cell of the urinary bladder is extremely uncommon, and up to date only 135 cases have been reported in word literature. Histologic, microscopic, and immunohistochemical characteristics are similar to oat cell carcinoma of the lung and other extrapulmonary oat cell carcinomas. We conclude this report with immunohistochemical study with PGP 9.5, neuron-specific enolase a synaptophisine. PMID- 10670133 TI - [Encrusted alkaline cystitis and malacoplakia]. AB - Urinary infection due to urea splitting bacteria leads to a rise in urinary pH, favouring the precipitation of calcium salts and struvita crystals. If deposited on the surface of a bladder with chronic inflammation or some other previous lesion, may produce an alkaline encrusted cystitis, now a rare condition. In the case here presented, occurred in a 69-year-old male. Corynebacterium urealyticum grown in the urine, and some foci of malakoplakia were found in the area of encrustation endoscopically excised. This case seems to be the third example of alkaline encrusted cystitis associated with malakoplakia reported in the bibliography. These two conditions share similar clinical signs and may probably have a common aetiopathogenesis. PMID- 10670132 TI - [Extragonadal germ cell tumor with "burned-out" phenomenon in the testis]. AB - The "burned-out" phenomenon in germ-cell neoplasias is defined by the presence of an extragonadal germ-cell tumour with no tumour at the testis level where a series of distinctive histological lesions can be detected indicative of the earlier presence of an already disappeared testicle tumoration. Extragonadal germ cell tumours with "burned-out" phenomenon show better evolution than their primary counterparts and are treated similarly to primary tumours of the testis. Currently, in the presence of retroperitoneal tumoration, a scrotal ultrasound study with high frequency transducers can lead to a suspected picture of tumoral involution. This paper contributes one retroperitoneal seminoma with "burned-out" phenomenon in the homolateral testis in a 35-year old patient. Available clinical and radiological criteria were enough to reach a suspected diagnosis. Homolateral orchiectomy and biopsy of retroperitoneal tumoration were performed, rounding treatment up with polychemotherapy. Evolution was good with immediate complete response. PMID- 10670134 TI - [Ask-Upmark kidney: description of a case and review of the literature]. AB - We present a 15-year-old male patient diagnosed histopathologically as suffering from Ask-Upmark kidney, in the absence of vesicoureteral reflux and with hypertension. The first clinical manifestation was completely atypical: right loin pain, with so many agudisation treated at our emergency serve that justified a thorough study. The pathogenesis of the Ask-Upmark kidney is still unknown; some authors defend the congenital malformation hypothesis, as it was first described in 1929, but there are groups who support the Ask-Upmark kidney as a form of reflux nephropathy. After our description we present a review of the literature. PMID- 10670135 TI - [Acute urologic symptoms associated with uterine myoma]. AB - While prevalence of uterine leiomyoma is high, its presentation affecting the urinary tract is uncommon. We contribute the cases of two adult women with symptoms of nephritic colic and urinary retention. Etiology was acute obstruction of the urinary tract due to previously asymptomatic urine myomas. Management in both patients was surgery, using hysterectomy to resolve the urinary obstruction. A brief review of the literature is included. PMID- 10670136 TI - [Tuberculous orchiepididymitis as clinical onset of human immunodeficiency virus (HIV) infection]. AB - The emergence of acquired immunodeficiency syndrome has changed the natural history of tuberculosis which has now become the second most common infection associated to human immunodeficiency virus infection. It is only rarely that a tuberculous infection has an urogenital location, and extrapulmonary locations are generally related to severe immunosuppression. This paper presents one case of tuberculous orchitis that presented as the clinical onset of acquired immunodeficiency syndrome. Discussion of the clinical evolution and the therapeutic approach that consisted in orchiectomy associated to treatment with tuberculostatics. PMID- 10670137 TI - [Penile fracture involving cavernous bodies and urethra]. AB - Reports on fracture of the penis are scarce in the spanish urological literature and in most cases injury appear limited to the corpora cavernosa. In this report a case of rupture of both corpora cavernosa and the urethra during sexual intercourse is presented. Early surgical treatment rendered good results. A literature review on currently recommended diagnostic and treatment practices is presented. PMID- 10670138 TI - [Hemorrhaging eso-gastro-duodenal ulcers: epidemiology and management. A multicenter prospective study]. AB - The aim of this study was to estimate the incidence, and to describe the characteristics and medical care in patients with bleeding upper gastrointestinal ulcers in the general population. PATIENTS AND METHODS: A study was performed over six months in 1996 in 4 French geographical areas: Finistere, Gironde, Seine Maritime, and the Somme (3 million people minimum 18 years). All public or private hospitals, and specialist gastroenterologists in private practice participated in the study, based on a standardized questionnaire. RESULTS: Over 6 months 793 patients with bleeding ulcers were identified, corresponding to 27 per 100,000 inh./year or 24,000 cases in France. Most patients were men (60%) and 40.1% were 75 years and older. The ulcer was oesophageal (6%), gastric (47%), or duodenal (69%). In 406 patients (51.2%) a chronic disease was present (cancer, cirrhosis, circulatory, respiratory or cardiac disease). In 237 cases (29.9%) the ulcer occurred in patients, 453 patients (57.1%) were admitted and 103 patients (13%) were managed as outpatients. Gastrotoxic drugs were taken by 349 patients (44%): non steroidal anti-inflammatory drugs (18.7%), aspirin (21.2%, including 2/3 with doses under 330 mg/day), corticosteroids (7.8%) and 24.3% had anticoagulant therapy. Patients were managed in university hospitals (39.3%), other public or non profit hospitals (44.2%) or private hospital (16.5%) with geographical differences between the 4 areas. Therapeutic endoscopy was performed in 16.9% and a surgical procedure was performed in 5.9%. The mortality rate (outpatients excluded) was 13.5% (n = 93), but only 2% (n = 16) of death were associated with a bleeding ulcer: mortality was higher in inpatients (24.1%) than in out patients (8.1%). A chronic disease was also associated with higher mortality (17.9% versus 8.1%). CONCLUSION: Bleeding ulcers are frequent and severe, especially in inpatients or associated with chronic conditions. A gastrotoxic drug used is found in about fifty percent of the cases. PMID- 10670139 TI - [Results and indications of lateral ileostomy functionally terminated in colorectal surgery]. AB - Loop ileostomy (LI) ensures fecal diversion to protect an anastomosis or anatomic colorectal or ano-perineal damage. The aim of this retrospective study was to evaluate loop ileostomy morbidity in emergency and planned colorectal surgery. PATIENTS AND METHODS: From 1991 to 1996, 145 loop ileostomies were performed in 139 patients, 77 men and 62 women with a mean age of 48.7 years (15-82). The etiology was a rectal tumor (cancer or large villous tumor n = 47), inflammatory bowel disease (n = 47, ulcerative colitis = 37 and Crohn's disease = 10) Familial Adenomatous Polyposis (n = 13) and other diseases (n = 32). 80% LI (n = 116) protected ileo-anal anastomoses (n = 46) colo-anal anastomoses (n = 45, 26 with colonic pouch), ileo-rectal anastomoses (n = 11) and other anastomoses (n = 15). 20% LI (n = 29) dysfunctional ano-perineal lesions (n = 8), anastomosis leak (n = 4) or distal bowel without intestinal resection (n = 17). RESULTS: 7 deaths were not stoma-related. 91% LI were closed after a mean diversion time of 3.6 months. LI closure was performed by a parastomal (n = 128) or laparotomy procedure (n = 4). Morbidity during LI diversion was observed in 24 patients (16.5%) 12 of whom (8.3%) were operated for small bowel obstruction (n = 6; 4.2%) stoma revision (n = 5; 3.5%) and prolapse (n = 1; 0.7%). 2 patients had peristomal skin excoriations, and 5 patients required readmission for dehydration due to high LI output. Morbidity after LI closure was observed in 12 patients (8.6%) 5 of whom were operated for anastomotic leak (n = 4) or small bowel obstruction (n = 1). Low morbidity and defunctioning efficiency confirm the indications for LI. LI is our first-line stoma in planned or emergency colorectal surgery. PMID- 10670140 TI - [Regulatory aspects of disinfection of endoscopes]. AB - The circular on the sterilization and the law of july 1, 1998 enact clearly that the medical devices that support steam sterilization must be sterilized with an organization of sterilization that ensure quality. Endoscope that enter normally sterile tissues should be subjected to a sterilization procedure before each use; if this is not feasible, they should receive high-level disinfection to destroy bacterial spores. The endoscope must be immersed for at least one hour in aqueous solution of 2% glutaraldehyde. This lengthened duration of processing must be integrate in the organization of medical department. The endoscopes not penetrating in a sterile cavity are disinfected with manual processing according to the protocol of the circular of april 1996, or with automated endoscope reprocessing machine. The different types of automated machine used to wash and disinfect endoscope must now answer to criterion concerning their design described in the circular of july 15, 1998. A significant work has to be made in hospitals to conform automated machine and procedure, to control the risk dependent with their use. PMID- 10670141 TI - [Traceability of sterilizable medical devices]. AB - Device traceability means recording and storing of data collected during the various steps of reprocessing of medical devices. We describe the purpose and methods to achieve device traceability. PMID- 10670142 TI - [Role of surgery in the treatment of refractory ascites in cirrhotic patients]. AB - Ascites, generally directly reflecting portal hypertension, is the commonest cause of hospitalisation in patients with cirrhosis. In almost 10% of patients with ascites, optimal medical treatment combining bed rest, salt and water restriction, and diuretic treatment, is unable to induce sodium excretion and decrease the volume of the ascites, corresponding to the definition of refractory ascites. In other cases, it is the treatment of ascites itself (salt and water restriction and diuretics) which induce complications: water and electrolyte disturbances, functional renal failure, encephalopathy, the development of which also corresponds to refractory ascites. The therapeutic armamentarium for the management of refractory ascites remains varied, with the use of aspiration of ascites with compensation, peritoneovenous shunts, transhepatic or surgical porto systemic anastomoses, and finally, liver transplantation. At the present time, each therapeutic measure must be taken while keeping in mind the possibility of subsequent liver transplantation and the potential risk of compromising liver transplantation by inappropriate treatments. In this context, the authors review and analyse the respective places of the various therapeutic modalities in the management of refractory ascites in cirrhotic patients. PMID- 10670143 TI - [Colorectal surgery]. PMID- 10670144 TI - [Hereditary colorectal cancer without associated polyadenomatosis]. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant condition in which affected individuals develop colorectal cancer or extracolonic cancer, most commonly endometrial, at an early age. Recent advances in molecular genetics have led to the identification of a germline mutation of DNA mismatch repair genes to be responsible for the majority of HNPPC cases. Since clinical screening of gene carriers can help to prevent cancer, it is important to devise strategies applicable to this syndrome. Recommendations for current management, especially screening and surgical treatment are under study, as they have not yet been clearly established. This paper reviews the clinical presentation, the molecular genetic diagnosis and therapeutic approaches of this syndrome including the controversies concerning prophylactic colectomy for cancer or gene carriers. PMID- 10670145 TI - [Hereditary colorectal cancer associated with polyposis syndromes]. AB - This study reviews different aspects of hereditary colorectal cancer associated with three polyposis syndromes: familial adenomatous polyposis, juvenile polyposis coli and Peutz-Jeghers syndrome. All these syndromes share some similarities: low incidence, autosomal dominant inheritance, genetic predisposition to colorectal cancer and/or other extracolonic cancers. Classical familial adenomatous polyposis is clinically defined by the presence of hundreds of adenomatous polyps in the colon and rectum, whereas less than 100 polyps are found in attenuated familial adenomatous polyposis. Without prophylactic colectomy, colorectal cancer develops inevitably by the age of 40. Restorative proctocolectomy with ileal anal-pouch anastomosis is the operation of choice in familial adenomatous polyposis. In juvenile polyposis coli, 50-200 hamartomatous polyps are found in the colon, rectum, stomach and small bowel. Life-time cumulative risk for colorectal cancer is estimated to be 50%. Prophylactic colectomy is required only in cases in which endoscopic surveillance is not able to control polyp development. Hereditary mixed polyposis syndrome is a variant form of juvenile polyposis coli, consisting of multiple mixed adenomatous, hyperplastic and hamartomatous polyps. Peutz-Jeghers syndrome is characterized by multiple hamartomatous polyps located in the small bowel, colon and stomach. Small bowel follow through and colonoscopy is advised for surveillance. Surgery is warranted only in cases of polyps larger than 1 cm. The causative genes of these syndromes have been cloned. Molecular genetic testing of affected and at risk individuals is proposed in order to advise surveillance and management. PMID- 10670146 TI - [Surgical anatomy of pelvic nerves]. AB - A good knowledge of the anatomy of the mesorectum and pelvic autonomic nerves allows the colorectal surgeon to reconcile both oncologic and functional results in rectal cancer excision. The author describes the anatomy of the systemic and autonomic pelvic nerves and describes techniques designed to avoid nerve damage during rectal cancer excision. PMID- 10670147 TI - [Surgery of rectal cancer: total exeresis of the mesorectum]. AB - The authors review the recent literature about total mesorectal excision for rectal cancer. They report the actual management of such patients. PMID- 10670148 TI - Total mesorectal excision with autonomic nerve preservation: a new foundation for the evaluation of multi-disciplinary adjuvant therapy in the management of rectal cancers. AB - Local and distant recurrence rates and disease-free and overall survival are markedly improved by total mesorectal excision, with little increase in morbidity, compared with other techniques of resection of rectal cancer. Adjuvant therapy is associated with significant morbidity and initial results suggest it may not be beneficial in the aggregate. Adjuvant therapy must be re-evaluated in trials using TME as standard operative technique. Different subgroups of patients, defined by clinical and pathological criteria will be best served by different forms of therapy and should be studied based on rates of local and distant recurrence. Selected groups of patients will be best served by undergoing no adjuvant therapy of any kind. PMID- 10670149 TI - [Preoperative radiotherapy of rectal cancer. The Lyons experience, 1985-1996. Prognostic study apropos of 312 patients]. AB - AIM OF STUDY: Preoperative radiotherapy is used increasingly in rectal cancer in Europe. This study is a retrospective analysis of a series of 312 patients with rectal adenocarcinoma treated by preoperative radiotherapy. MATERIAL AND METHOD: From 1985 to 1996, 312 patients were included in this study. Preoperative staging was: T2: 83, T3: 192 et T4: 21. On digital rectal examination, 25 patients were classified as N1. Endorectal sonographic staging was: uT1: 3, uT2: 77, uT3: 163, uN0: 122, uN1-2: 127. After surgery, pathological staging was: pT0: 43 (14%), pT1 24, pT2: 81, pT3: 151, pN0: 229, pN1-2: 81. Radiotherapy was delivered to the posterior pelvis with an accelerated schedule 39 Gy/13 fractions/17 days with x 18 MV. RESULTS: Median follow-up is 54 months. For pM0 patients (297 patients), the overall 5-year survival rate is 67%. Local failure rate is 9%. Since 1986, the rate of sphincter saving surgery is close to 65%. Various parameters related to the tumor were found to be significant prognostic factors on multivariate analysis in relation to 5-year overall survival rate: the T stage as judged by digital rectal examination and endorectal sonography, the N stage as evaluated on digital rectal examination but not with endorectal sonography. Pathological examination of the operative specimen retains a very strong prognostic value for pT and pN. CONCLUSION: Pathological examination of the specimen of rectal carcinoma retains a very strong prognostic value after preoperative radiation therapy. Endorectal sonography is of interest to evaluate T staging of the tumor but is not reliable for N stage. PMID- 10670150 TI - [Established data and practical recommendations concerning pre- and postoperative chemotherapy of rectal cancer]. AB - Adjuvant chemotherapy appears to be active in stage II-III rectal cancers; the NSAPB R01 trial demonstrated a survival advantage for patients receiving chemotherapy using the MOF protocol and 3 meta-analyses are in favor of the efficacy of adjuvant chemotherapy in rectal cancer. Three randomized trials have also demonstrated that combinations of radiation and chemotherapy are superior to surgery alone or adjuvant radiotherapy and demonstrated the major role of systemic chemotherapy combined with radiotherapy. However this efficacy of adjuvant chemotherapy alone or combined with radiation therapy is still debated and specific trials must be conducted to test the value of chemotherapy using more active regimens than those previously tested and taking into account the quality of surgery and radiotherapy; such trials are in progress, especially the trial conducted by the EORTC and the FFCD. The efficacy of neoadjuvant chemotherapy has never been clearly demonstrated, although a combination of radiotherapy and chemotherapy as first-line treatment for locally advanced rectal cancer and in the case of synchronous metastasis seems to facilitate surgical resection. It is a reasonable and tolerable approach with manageable toxicity which gives substantial results in 2/3 of patients. This strategy also allows better selection of patients likely to benefit from surgical resection of their primary tumor and in some cases of their synchronous metastases. However, the efficacy of perioperative treatments should not decrease the quality of the surgical resection and especially mesorectal excision as well as the need for high quality pathological examination which must be very thorough with analysis of a sufficient number of lymph nodes. The efficacy of combined treatment in advanced rectal cancers is a major argument in favor of the multidisciplinary coordination required for optimal treatment of patients with rectal cancer. PMID- 10670151 TI - [Role of subtotal/total colectomy in emergency treatment of occlusive cancer of the left colon]. AB - PURPOSE: The aim of this study was to evaluate the results of management of acutely obstructed carcinoma of the left colon by emergency subtotal/total colectomy (STC) with immediate anastomosis without diversion. METHODS: STC was performed in 60 consecutive patients (mean age 72 years). Inclusion criteria were reasonable operative risk, resectable acutely obstructed carcinoma, massively distended colon of dubious viability, signs of impending cecal perforation. RESULTS: Postoperative mortality was 6.6% (n = 4): 3 patients over 85 years of age died postoperatively as a result of cardiopulmonary complications; an 83 year old female died as a result of an anastomotic dehiscence. Morbidity was 10% (n = 6) including one fistula which recovered without surgery. There were 5 synchronous colon cancers. Six months after surgery, the mean daily stool frequency was 2 after STC, and 3 after TC. CONCLUSION: Emergency STC achieves one stage relief of bowel obstruction and tumor resection by encompassing a massively distended and fecal-loaded colon with ischemic lesions, ensures restoration of gut continuity via a "safe" anastomosis and removes a possible synchronous carcinoma. PMID- 10670152 TI - [Left colectomy with immediate anastomosis in emergency surgery]. AB - PURPOSE OF THE STUDY: A retrospective study of our experience with one-stage left colectomy for acute diverticulitis and obstruction with a review of the literature to more clearly define the indications of this procedure. PATIENTS AND METHODS: 30 patients were operated for acute diverticulitis (group 1) and 47 for obstruction (group 2). Only 7 patients (23%) of group 1 had an intraoperative colonic lavage while this was performed for all the patients of group 2. RESULTS: The postoperative morbidity and mortality for the patients of group 1 and 2 were 37 and 28%, and 7 and 11% respectively. None of the patients of group 1 had clinical anastomotic leak, while this occurred in 2 patients (4%) of group 2. The mean hospital stay was 26 days for patients of group 1 and 17 days for patients of group 2. CONCLUSIONS: Bowel obstruction should be treated by one-stage left colectomy and intraoperative colonic lavage for patients with low anaesthetic risks (ASA 1 and 2). Immediate anastomosis protected by colostomy or ileostomy could be proposed for patients with an intermediate risk (ASA 3). Patients with acute diverticulitis and a localized abscess or peritonitis should be treated with one-stage colectomy; an immediate protected anastomosis could be performed in patients with generalized purulent peritonitis while a Hartmann's type colectomy should be the reasonable option for fecal generalized peritonitis. Intraoperative colonic lavage does not seem mandatory. PMID- 10670153 TI - [Self-expandable metal stent in the treatment of obstructive cancer of the left colon. Preliminary results and review of the literature]. AB - AIM: To report our preliminary experience with self-expandable metal stent in the treatment of acute malignant obstruction of the left colon and to review the literature on this specific subject. PATIENTS AND METHODS: From March to September 1999, 8 consecutive patients with a mean age 71 were admitted as an emergency for acute malignant obstruction of the left colon. A self-expandable metal stent was inserted under radioscopic and, in 4 cases, endoscopic guidance. The patients then underwent bowel preparation before operation, if required. RESULTS: There was no mortality. Bowel preparation was satisfactory in 6 cases. Complications occurred in 1 patient, who was operated on day one for peritonitis due to perforation of the tumour by the prosthesis inserted after dilatation. Another six patients were operated: 2 had resection followed by anastomosis; 3 had resection and anastomosis protected by ileostomy; 2 had Hartmann's procedure. The last patient retained the prosthesis as palliation. In the literature, self expandable metal stent application in obstructed carcinoma of the left colon gives satisfactory results. CONCLUSION: Based on our experience and a review of the literature, we provide practical recommendations when inserting self expandable metal stents for acute malignant left colonic obstruction. PMID- 10670154 TI - [Colonic diverticulosis and laparoscopy. Analysis of a series of 60 cases]. AB - AIM: This is a retrospective analysis of a series of 60 cases diverticular disease. MATERIAL AND METHOD: From May 1991 to April 1999, 60 laparoscopic colorectal resections were performed for diverticulitis. RESULTS: Conversion to a classical procedure was necessary in 3 patients (5%). There was no mortality and 9 postoperative complications (3 reoperations). The mean length of hospital stay was 9 days, and 6.3 days for patients in whom surgery was performed after January 1998. CONCLUSION: Laparoscopic surgery for diverticular disease is associated with acceptable morbidity and mortality rates and a short median postoperative stay. PMID- 10670155 TI - [Laparoscopic ileo-colic resection in Crohn's disease]. AB - The objective was to evaluate the reliability and safety of laparoscopic ileocolic resection for Crohn's disease. PATIENTS AND METHODS: From June 1995 to February 1999, 40 patients underwent a laparoscopic ileocolic resection for Crohn's disease. Fistulizing disease, phlegmons and patients with previous laparotomy were excluded. Early morbidity, postoperative comfort and clinical recurrence were rates evaluated. RESULTS: No intra-operative incident or conversion occurred. Mean operating time was 163 min. Complications occurred in three patients: 1 pelvic hematoma with super-infection, 1 protracted ileus (7 days), 1 venous thrombosis. Opiate analgesics were used for a mean period of 3.1 days. Delay before bowel movements was 3.2 days. Post-operative hospital stay was 8 days. Mean size of the wound was 4.1 cm. Twelve patients (30%) developed long term clinical recurrence; the mean disease-free interval was 10 months. No patient required secondary re-operation. CONCLUSION: Laparoscopic ileocolic resection was reliable and safe in the treatment of Crohn's ileal strictures. The possible role of this method in the treatment of fistulizing disease or recurrence has to be evaluated. PMID- 10670156 TI - [Spontaneous colonic ischemia]. AB - Spontaneous ischemic colitis is a frequent disease, affecting mostly elderly people and predominantly localised on the left and sigmoid colon. It is due to alterations of small vessels of the colon and/or modifications of the splanchnic blood flow. Spontaneous evolution is generally favorable with restitution ad integrum of the disease colon. The diagnosis of ischemic colitis is established from the clinical picture, CT-scan and colonoscopy. Surgery, under the form of colonic resection, is required in 15% of cases approximately, in case of colonic necrosis. PMID- 10670157 TI - [Concerning "Intraperitoneal polyglactin 910 absorbable prosthesis: a risk-free preventive method?"]. PMID- 10670158 TI - [Interest in ophthalmologic examination for familial adenomatous polyposis]. AB - We describe the clinical history of four patients belonging to a familial adenomatous polyposis (FAP) family. We stress the importance of a fundus examination for the screening of carriers of the gene responsible for familial adenomatous polyposis. PMID- 10670159 TI - [Congenital toxoplasmosis: presentation of a case]. AB - We report a case of a 4 months old boy with esotropia of the left eye with large bilateral chorioretinal toxoplasmic macular scars. Chorioretinal scars are the most common eye finding in congenital toxoplasmosis and are often located in the macular region. Most infants with congenital infection are asymptomatic at birth but will develop retinal and/or neurologic damage later in life with consequent loss of vision. A routine examination of the fundus and computed tomography of the head can be negative. Serologic testing is essential for the diagnosis and the follow-up of the infection. Every infant with evident or suspected congenital infection by Toxoplasma gondii must be treated by Pyrimethamine, Sulphadiazine and Folinic Acid during at least the first year of life with regular serologic testing and ophthalmologic examination. Neurologic outcome is better with treatment and the risk of chorioretinitis seems reduced. PMID- 10670160 TI - Silicone oil vs. gas for the treatment of full-thickness macular hole. AB - The purpose of this study is to evaluate the anatomic and visual outcomes, as well as the complications, of macular hole surgery with SF6-gas tamponade versus silicone-oil tamponade. Fifty-four (54) eyes with idiopathic macular hole underwent vitrectomy and peeling of the internal limiting membrane (ILM) around the hole. Nineteen (19) eyes were treated with SF6-gas tamponade (group 1) and the other thirty-five (35) eyes with silicone-oil tamponade (group 2). An excellent anatomic success rate was obtained in both groups (94.7% in group 1 and 97.1% in group 2). Nevertheless, the postoperative visual acuity (VA) in the group treated with silicone-oil tamponade was significantly better than in the group treated with gas tamponade (P = 0.0217). Forty-seven (47) of the eyes in group 1 and 74% in group 2 achieved a VA = 0.4 or better. The most frequent potentially vision threatening complication we observed was RPE alterations in 35% of the eyes in group 1 and in only one eye in group 2. None of the eyes developed a retinal detachment during the follow-up period. In conclusion, the treatment of idiopathic macular holes by vitrectomy and ILM peeling provides a very good anatomic success rate. An excellent recovery of visual acuity, up to 1.0, was more frequently observed in the group treated with silicone oil tamponade. PMID- 10670161 TI - [Placoid pigment epitheliopathy and cerebral vasculitis: a clinical case]. AB - We report the case of a 21 year old man who has severe headache and blurred vision since 2 weeks. Ophthalmologic examination discloses typical lesions of acute posterior multifocal placoid pigment epitheliopathy and an homonymous right inferior quadrantanopsia. An inflammatory syndrome and a cerebrospinal fluid lymphocytosis are found. Cerebral imagery is normal. Headache improves only with corticotherapy. We conclude that the neurological attack associated with this acute posterior multifocal placoid pigment epitheliopathy is most likely due to a cerebral vasculitis. PMID- 10670162 TI - Radiation-associated choroidal neovasculopathy, exudative detachment and neovascular glaucoma. A case report. AB - Radiotherapy remains a controversial type of therapy for subfoveal neovascularization. Recently a peculiar pattern of neovascular growth of the irradiated choroidal neovascular membrane has been described. This evolution may be associated with extensive exudative reaction. In one of our patients with this complication, the disease progressed to a total exudative retinal detachment and neovascular glaucoma. PMID- 10670163 TI - Target pressures in glaucoma. AB - Despite exciting progress in the field of neuro-protection, lowering the intraocular pressure is still the only available option to treat glaucoma patients. The level to which the intraocular pressure should be lowered is different for each individual patient. The formula proposed to calculate the "target pressure" takes into account the pressure at which the glaucomatous damage presumably occurred (the "maximum pressure") and the risk of future damage. This target pressure should be re-evaluated periodically. PMID- 10670164 TI - Acanthamoeba keratitis: a review. AB - Acanthamoeba keratitis is caused by protozoa and characterised by a protracted course. All patients presenting with a therapy-resistant keratitis, even non contact lens wearers, should be examined for the presence of Acanthamoeba by means of specific cultures, histopathological stainings and--if necessary--a corneal biopsy. The combination of clinical signs, such as excessive pain, a radial keratoneuritis and in a later phase a stromal ring infiltrate, together with a suggestive history (contact lenses, polluted water) is an important factor for the early diagnosis. Because of improved clinical detection and earlier diagnosis, the infection can often be controlled with a combination therapy of polyhexamethylene biguanide or chlorhexidine with propamidine and neomycine. This results in a better visual prognosis and a decreased need for therapeutic keratoplasty. PMID- 10670165 TI - Comparison of the effect of Healon vs. Viscoat on endothelial cell count after phacoemulsification and posterior chamber lens implantation. AB - To evaluate the protective effect of 1% sodium hyaluronate (Healon) vs. a mixture of 4% chondroitin sulfate and 3% sodium hyaluronate (Viscoat) on the central corneal endothelium during surgery, we prospectively examined 20 eyes of 20 patients who underwent clear corneal phacoemulsification and implantation of an intracapsular posterior chamber intraocular lens (PC-IOL) with either Healon (10 eyes) or a combined use of Viscoat and Healon (10 eyes) as viscoelastic material. We compared the central endothelial cell counts, recorded by specular microscopy, preoperatively and 6 weeks postoperatively. Our results suggest that Viscoat offers no significant better endothelial cell protection during phacoemulsification than Healon. PMID- 10670166 TI - A one year experience with the multislice helical CT. AB - New detector morphology and increased computer power have led to a second leap in CT technology. With multislice helical CT, four slices per rotation are reconstructed resulting in a 3 to 6 times reduction of the examination time. During our one-year experience with the multislice CT, we have learnt that the major advantage of the technique is the possibility to cover a large volume with thin slices. Combination of both makes it possible to scan an entire anatomic region during the optimal enhancement phase (after i.v. contrast injection) with a high spatial resolution. New diagnostic possibilities can be explored by CT together with improved and more detailed post-processing tools. PMID- 10670167 TI - [Alveolar proteinosis: signs and prognosis using high-resolution computed tomography in 5 patients]. AB - Pulmonary alveolar proteinosis is a rare idiopathic diffuse airspace disease characterized by intraalveolar accumulation of large quantities of lipoproteinaceous material. The clinical presentation and course are variable and the definite diagnosis is made by biopsy or broncho-alveolar lavage (BAL) that also constitutes the unique and empiric treatment. We report the extremely typical HRCT features of the disease found in a series of five patients diagnosed in our institution over a ten-year period. The HRCT signs and their evolution are discussed. A continuous spectrum of findings going from isolated ground glass opacities to lobular consolidation is found, but the most common and invariably present pattern consists of ground glass areas with superimposed smooth septal thickening; these areas have a patchy or geographic distribution--often termed the "crazy-paving" pattern--and are unlike pulmonary findings in any other disease. PMID- 10670168 TI - An undescribed coexistence of a subserosal exophytic gastric leiomyoma with a serous microcystic pancreatic adenoma. AB - We present the case of a 66-year-old woman with complaints of odynophagia but normal gastro-oesophagoscopic findings. On computed tomographic examination, a presumptive diagnosis of a cystic liver tumor and a microcystic pancreatic adenoma was made. Histopathological examination of the hepatic mass, however, revealed a subserosal exophytic gastric leiomyoma. Considerations to avoid the potential pitfall of diagnosing an exogastric leiomyoma as a liver tumor are emphasized. Moreover, the unique association of an exogastric leiomyoma and a serous microcystic pancreatic adenoma is discussed. PMID- 10670169 TI - Leptomeningeal metastases from ethmoid sinus adenocarcinoma: clinico-radiological correlation. AB - A patient with sinonasal adenocarcinoma is presented with leptomeningeal metastases affecting multiple cranial nerves and spinal nerve roots. Head and neck cancer is known to be an extremely rare source for leptomeningeal metastatic spread. The cranial nerves, the spinal cord and roots and the cerebral hemispheres can be affected in case of leptomeningeal metastatic spread. Examination of the CSF is the hallmark of the diagnosis if leptomeningeal metastatic spread is suspected, but this case illustrates that the combination of specific clinical features on one hand and specific lesions on the Gd-enhanced T1 weighted MRI study on the other hand is reliable enough to make a presumed diagnosis if the CSF analysis remains negative. We suggest that in our patient direct leptomeningeal spread occurred through the cribriform plate to the CSF, followed by further spread in a gravity dependent way. PMID- 10670170 TI - Confrontation of mammography systems in flanders with the European Guidelines for Quality Assurance in mammography screening. Analysis of initial results. AB - From May 1997 to April 98, 30 radiological centers made an agreement with the Leuvens Universitair Centrum voor Kankerpreventie (LUCK) for the work out of physical and technical quality control in mammography screening. A protocol was used based on the European Guidelines for Quality Assurance. The reports of all 30 acceptance tests were retrospectively reviewed. The following parts showed to be most critical: alignment of radiation field and film in the bucky, tube voltage precision, automatic exposure controller, average optical density of a standard exposure, dark room and its safe lights. The mean film gradient was in the majority of the centers higher than what is prescribed in the European document. This is acceptable and even desirable whenever daily quality control shows that the development system is sufficiently stable. Although it is difficult to compare the scores for the specific tests with results in other countries, there is evidence that the tendencies are very similar. PMID- 10670171 TI - Tumours of the oropharynx and oral cavity: perineural spread and bone invasion. AB - Clinical examination of the oral cavity and oropharynx provides essential information in the assessment of neoplastic lesions. A precise evaluation of their deep spread along the most common growth pathways can be achieved by imaging, ranging from the basic, but nowadays incomplete, information of conventional X-ray, to the sophisticated details obtained by MR. Three oncological questions must be faced: the three dimensional evaluation of primary tumour spread; the assessment of nodal involvement; the post-treatment survey with the early detection of local recurrences, during the follow up. Either CT or MR accurately assesses the deep extension of neoplasms, nevertheless, the most cost-effective protocol is provided by a combination of CT and ultrasound (staging respectively T and N). MR is the technique of first choice when an infiltration of the base of the tongue or perineural spread is suspected, because of its superior ability to detect muscular invasion and segmental abnormalities of cranial nerves. Bone involvement can be adequately showed by MR not only because focal erosions of the cortical rim are well demonstrated, but also by means of the early demonstration of bone marrow abnormalities. Moreover, MR plays an essential role during the follow up, as it is the only morphological imaging technique permitting to differentiate recurrent tumour and necrosis from scar tissue. PMID- 10670172 TI - Cross-sectional anatomy of the facial nerve. AB - The length and complexity of the anatomical course of the facial nerve explains the difficulty of its accurate morphologic evaluation. CT and MR appear to be complementary techniques to precisely depict the nerve from its pontine origin to the parotid gland. Anatomical variations exist in length or thickness of all intrapetrous segments or as frequent dehiscences which can lead to false positive results or at the opposite falsely negative diagnoses. Close relations with the antero-inferior cerebellar artery in the intracisternal and intracanalicular segments must be known. Gadolinium enhancement is usual in the fallopian canal with variable intensity and thickness and should be differentiated from pathological enhancement. Finally the intrapetrous course of the chorda tympani can be precisely displayed on CT in the intra-osseous canal and in the middle ear near the ossicles. PMID- 10670173 TI - Aorto-enteric fistula. PMID- 10670174 TI - Clinical aspects of pulmonary sarcoidosis. AB - Pulmonary sarcoidosis has a variable natural course from an asymptomatic state to a progressive life-threatening condition. Radiographic abnormalities are frequently an important clue to the diagnosis. The diagnosis usually requires a tissue biopsy that demonstrates noncaseating granulomas that cannot be ascribed to another clinical condition. The lung may be biopsied, but extrapulmonary sites may be biopsied for the diagnosis if such sites are involved with disease. When the lung is biopsied, a transbronchial lung biopsy with a flexible bronchoscope is the procedure of choice, even if the chest radiograph shows thoracic adenopathy alone without obvious parenchymal infiltration. On occasion the diagnosis can be made on clinical grounds without biopsy when the presentation is highly specific for sarcoidosis, such as Lofgren's Syndrome. Treatment for pulmonary sarcoidosis has not been standardized. Since many patients have spontaneous remissions and the benefits of therapy do not affect the long-term outcome, therapy is reserved for patients with severe or progressive pulmonary symptoms and/or pulmonary dysfunction. Corticosteroids are the primary therapy for pulmonary sarcoidosis. Corticosteroid therapy involves six phases: initial high dose, taper to a maintenance dose, a maintenance dose, taper off corticosteroids, monitor off therapy, and treatment of relapse if it occurs. PMID- 10670175 TI - Alzheimer's disease in South Carolina, 1998. PMID- 10670176 TI - Curricular renewal for the new millennium at the Medical University of South Carolina College of Medicine. AB - The MUSC College of Medicine is engaged in a curricular renewal process that emphasizes increased integration of the basic and clinical sciences throughout all four years of the curriculum, more self-directed learning, and earlier patient contact for students. Several basic science courses have been modified and a new "Doctoring Curriculum" has been introduced to develop students' clinical skills and preparation for medical practice. Changes to the third year of the curriculum include the new Deans' Rural Primary Care Clerkship. Other third-year curricular changes include small-group case discussion sessions that emphasize the integration of basic and clinical sciences in medical practice, and the incorporation of nutrition throughout all fours of the curriculum. The changes described in this manuscript are designed to address a wide range of educational needs of future physicians, including the acquisition of the attributes endorsed by the AAMC MSOP--altruism, knowledge, skillfulness and dutifulness. This new curriculum will evolve over time and the goal remains to help equip future physicians with the requisite knowledge, skills and attitudes for medical practice in the new millennium. PMID- 10670177 TI - Y2K.1. The tree of knowledge and the tree of life. PMID- 10670178 TI - [Levels of total plasma homocysteine after methionine intake in a group of healthy subjects in Northern + Italy. Relationship with age, gender, and nutritional status]. PMID- 10670179 TI - [Risk factors of cardiovascular pathology after kidney transplantation. New perspectives]. PMID- 10670180 TI - [I/D polymorphism of the ACE gene and A1166C of the AT1R gene as risk factors for restenosis after coronary angioplasty]. PMID- 10670181 TI - [The genes that codify angiotensin converting enzyme and type 1 receptor for angiotensin II have a different effect on longevity]. PMID- 10670182 TI - Molecular evidences for a hypercoagulable state in non insulin diabetes mellitus patients with a history of thrombotic episodes. PMID- 10670183 TI - [Vascular disease and asymptomatic diabetic patients]. PMID- 10670184 TI - [Apolipoprotein-E4, lipid profile and vascular dementia]. PMID- 10670185 TI - [Vascular risk factors, cognitive status, and affective tone in 2 groups of elderly patients]. PMID- 10670186 TI - [Thrombophilic states. Prevalence and clinical correlations]. PMID- 10670187 TI - [Lp(a): a new risk factor for deep venous thrombosis?]. PMID- 10670188 TI - [The C1166 allele of the AT1R gene associated with ACE DD phenotype increases the risk for deep venous thrombosis]. PMID- 10670189 TI - [Deep vein thrombosis in patients undergoing medical treatment. One-year experience at a rehabilitation hospital]. PMID- 10670190 TI - [Important risk factors for venous thromboembolism. Analysis of 758 patients studied at a thrombosis center]. PMID- 10670191 TI - [Diagnostic accuracy of various methods for the measurement of D-dimers in patients with suspected deep venous thrombosis]. PMID- 10670192 TI - [Asymptomatic deep vein thrombosis in the bedridden elderly. Role of D-dimer as screening test]. PMID- 10670193 TI - [Recurrences and new vein thrombotic events during oral anticoagulant therapy in patients treated for previous vein thromboembolism. Data from the literature and results of the ISCOAT study]. PMID- 10670194 TI - [Pretreatment with monoclonal antibody anti-CD18 prevents the trans-cellular biosynthesis of sulfidopeptide-leukotrienes in vitro and in vivo and protects from the subsequent functional changes in the isolated rabbit heart]. PMID- 10670195 TI - [Marfan's syndrome. Clinical and molecular characterization of 51 Italian patients]. PMID- 10670196 TI - [Hyperhomocysteinemia and abdominal aortic aneurysm]. PMID- 10670197 TI - [Thoracic aortic aneurysm. New evidence for fibrillin-1 involvement]. PMID- 10670198 TI - [Changes in fibrinolysis after primary PTCA]. PMID- 10670199 TI - [Persistent generation of thrombin after coronary angioplasty]. PMID- 10670200 TI - [Effects of an angiotensin II receptor antagonist (valsartan) on macro- and microcirculatory hemodynamics of the arm in hypertensive patients]. PMID- 10670201 TI - [Passive smoking and endothelial dysfunction]. PMID- 10670202 TI - [Does the presence of obliterative arteriopathy of the legs change the long term mortality and survival of patients surgically treated for abdominal aortic aneurysm?]. PMID- 10670203 TI - [Preliminary experience in the treatment of thoracic aortic aneurysm with covered self-expandable endoprosthesis]. PMID- 10670204 TI - [Aneurysm of the lumbar aorta treated with a transluminal approach. Experience at one center in 24 months]. PMID- 10670205 TI - [Use of endoprosthesis in aortoiliac aneurysms]. PMID- 10670206 TI - [Aneurysm of popliteal artery caused by extrinsic compression]. PMID- 10670207 TI - [Surgical treatment of aneurysm of the popliteal artery. Immediate and long-term results]. PMID- 10670208 TI - [Significance of early diagnosis in vascular prosthetic infections]. PMID- 10670209 TI - [Late arterial thrombolysis. Indications, methods, and results of our experience]. PMID- 10670210 TI - [Structural changes of the radial artery in patients with peripheral atherosclerotic arteriopathy]. PMID- 10670211 TI - [Peripheral arteriopathy in pseudoxanthoma elasticum]. PMID- 10670212 TI - [Atherogenic risk factors in patients with obliterative arteriopathy of the legs]. PMID- 10670213 TI - [Prevalence, topographic distribution, and pathogenesis of macro- and microangiopathic vascular lesions in patients with diabetes mellitus. Retrospective study]. PMID- 10670214 TI - [Relationship between hyperhomocysteinemia and endothelial activation in patients with obliterative arteriopathy of the legs]. PMID- 10670215 TI - [Hyperhomocysteinemia and endothelial damage in abdominal aortic aneurysm]. PMID- 10670216 TI - [Cardiopulmonary baroreceptors activity and skin microcirculation in patients with peripheral obliterative arteriopathy]. PMID- 10670217 TI - [Prognostic value of transcutaneous oximetry in critical ischemia]. PMID- 10670218 TI - [Proposed guidelines for the correct application of ultrasonography methods in the diagnosis of deep venous thrombosis of the legs]. PMID- 10670219 TI - [Sural vein thrombosis and recurrence prevention. Which therapy?]. PMID- 10670220 TI - [Radiology in the prevention of pulmonary embolism. Vena cava filters]. PMID- 10670221 TI - [Home use of an ambulatory monitor for the determination of prothrombin time (coaguchek) in patients treated with oral anticoagulants. Multicenter prospective study]. PMID- 10670222 TI - [Venous and arterial thromboembolism: autoimmune syndrome with heparin-induced thrombocytopenia. Unusual severe adverse reaction or the tip of the iceberg?]. PMID- 10670223 TI - [Quantification of venous reflux of the legs with scanner duplex method]. PMID- 10670224 TI - [Role of color Doppler ultrasonography in the study of venous ulcer of the leg]. PMID- 10670225 TI - [ULTRO protocol in the study and treatment of stasis ulcer of the leg]. PMID- 10670226 TI - [Changes of endothelium-dependent vasodilation in healthy subjects with familial atherosclerosis of the supra-aortic branches]. PMID- 10670227 TI - [Dermal microcirculatory response to various stimulation tests assessed with Doppler laser]. PMID- 10670228 TI - [Quantitative assessment of irrigation dermal microcirculation with computerized biomicroscopy]. PMID- 10670229 TI - [Endothelial function of dermal microcirculation in patients with essential arterial hypertension]. PMID- 10670230 TI - [Site specificity of biomicroscopic pattern in diabetic patients]. PMID- 10670231 TI - [Local anesthesia in carotid surgery]. PMID- 10670232 TI - [Indications and results of carotid TEA in the 90's]. PMID- 10670233 TI - [Perioperative myocardial ischemia in patients treated with carotid surgery. Impact of the type of anesthesia (locoregional vs general]. PMID- 10670234 TI - [Selective use of shunts in carotid surgery. Personal experience]. PMID- 10670235 TI - [Carotid stenting. Immediate and long-term results]. PMID- 10670236 TI - [Incidence of atherogenic risk factors, carotid atherosclerosis, and eventual correlations between etiology and clinical outcome in a group of patients with cerebral ischemia. Retrospective study]. PMID- 10670237 TI - [Carotid atherosclerosis and juvenile stroke. First results of a longitudinal study]. PMID- 10670238 TI - [Helicobacter pylori, Chlamydia pneumoniae, reactive C protein, and cerebrovascular diseases]. PMID- 10670239 TI - [In situ activation of metalloproteinases by the fibrinolytic system as pathogenetic mechanism of rupture of the atherosclerotic plaque]. PMID- 10670240 TI - [Ultrasonography tissue characterization of the carotid wall with integrated backscatter signal analysis. A new alternative for the early diagnostic assessment of atherosclerosis]. PMID- 10670241 TI - [Thickness of the intima media and coronary disease]. PMID- 10670242 TI - [Doppler color ultrasonography in the assessment of epiaortic vessels in giant cell arteritis]. PMID- 10670243 TI - [Transcranial color coded sonography (TCCS) in the diagnosis and follow up of spontaneous dissection of the internal carotid artery]. PMID- 10670244 TI - [Comparison between L-propionyl carnitine and physical-rehabilitative exercise in diabetics with obliterative arteriopathy of the legs (Fontaine's stage IIa)]. PMID- 10670245 TI - [Alprostadil combined with antithrombotic drugs. Effects on hemorrhage time]. PMID- 10670246 TI - [Effects of infusion treatment once a day for 4 weeks with alprostadil-alpha cyclodextrin on blood levels of endothelin-1 in patients with chronic obliterative arteriopathy of the legs at Leriche-Fontaine stage 2]. PMID- 10670248 TI - Short term treatment with prostanoids in critical limb ischaemia: three months follow-up. PMID- 10670247 TI - [Shunt effect in iloprost treatment of obliterative arteriopathy]. PMID- 10670249 TI - [Treatment with prostanoid drugs. Analysis of 131 cases]. PMID- 10670250 TI - [Medical treatment of peripheral arteriopathies with alprostadil-alpha cyclodextrin. Comparative analysis at national level of the expense/reimbursement ratio]. PMID- 10670251 TI - [Gene therapy of critical ischemia of the legs. Improvement of genetic transfection efficiency of endothelial cells in vitro, using cationic liposomes and protamine sulfate]. PMID- 10670252 TI - Intimal-medial thickness of the common carotid arteries and lower limbs atherosclerosis in the elderly. AB - BACKGROUND: In the present study, the authors consider the possible association between intimal-media thickness of the common carotid arteries and lower limb atherosclerosis, in a group of elderly patients; the authors also consider the presence of cardiovascular risk factors. METHODS: B-mode ultrasound measurement of the intimal-media thickness of the common carotid arteries was performed on 80 subjects. Lower limbs atherosclerosis was defined as the presence of intermittens claudicatio and/or ankle-arm index < 0.9. Baseline clinic examination and blood tests were performed in all subjects to consider the presence of cardiovascular risk factors. Linear regression analysis was used to assess the linear relationship between intimal-media thickness of the common carotid arteries and lower limb atherosclerosis. RESULTS: Linear regression analysis showed a statistical association between increased values of intimal-media thickness of the common carotid arteries and lower limb atherosclerosis. Sixty-four (80%) subjects presented one or more associated cardiovascular risk factors. CONCLUSIONS: The results of the the present study confirm that intimal-media thickness of the common carotid arteries is a marker for the identification of generalized atherosclerosis and may be useful for the identification of subjects, even at early stages, at risk of cardiovascular diseases. PMID- 10670253 TI - Recurrent aneurysms: late complication for patients previously submitted to graft replacement for abdominal aortic aneurysm. AB - BACKGROUND: The possibility to perform easy and cheap ultrasonographic examinations of the abdomen allows us to face the problem of possible development of new paraanastomotic aneurysms or aneurysms on native arteries in subjects already submitted to endoaneurysmectomy of the abdominal aorta. The idea of planning accurate programs of ultrasonographic follow-up of operated patients is sustained by the hypothesis that the aortic aneurysm is an expression of a systemic disease due to a connectival defect of genetic nature. METHODS: Two different follow-up experiences are described: the first was a simple follow-up based only on clinical evaluation of patients previously submitted to aortic surgery in the last ten years at the Department of General and Cardiovascular Surgery of the University of Milan, and the second experience a planned follow-up programme based on instrumental evaluation of the patients. RESULTS: Thirty-one cases of recurrent aneurysms of native arteries in 13 subjects already operated for AAA from 3 to 13 years before are described. Despite the patients were evenly inserted in a clinical follow-up, as a matter of fact, 25 of these lesions were detected during occasional investigations performed for other reasons or for rupture, while during the last two years, the planned ultrasonographic follow-up of 95 patients detected 6 new aneurysms. CONCLUSIONS: Incidental detections of new paraanastomotic true aneurysms or ectasia of native arteries far from the graft are more and more frequent in patients already submitted to aortic replacement for abdominal aneurysm. In the international literature only few papers have been published about perspective studies based on instrumental follow up of operated patients. The planning of such controls is mandatory, at present, also in order to evaluate the clinical development of aneurysmatic disease. PMID- 10670254 TI - Relationships between serum hyperhomocysteinemia and carotid atherosclerosis in geriatric patients. AB - BACKGROUND: Evaluate the relationships between serum total hyperhomocysteinemia and carotid atherosclerosis. METHODS: The 102 consecutive patients over the age of 65 epiaortic vessels were examined by means of a high-resolution echo-Doppler and Doppler cw. Depending on the size of the atherosclerotic plaques in the carotid the patients were divided into two groups: a) patients with lesions at high thromboembolic risk (heterogeneous and/or ulcerated plaques, hemodynamically significant stenoses > 70%); b) individuals with low risk carotid involvement (IMT normal and/or thickened by > 0.8 mm and/or lumen stenosed by homogeneous plaques < 70%). The quantitative assay of serum total homocysteinemia (tHCY) was carried out by means of an immunofluorescent method (FPIA). Depending on their tHCY values, the patients were broken down into those with normal tHCY (cut-off 14 mumol/l for the women and 16 mumol/l for the men; 62 patients, M/F = 34/28, mean age 71 +/- 4); patients with mild hyperhomocysteinemia (tHCY 14-18 mumol/l for the women; 16-20 mumol/l for the men; 19 patients, M/F = 11/8; mean age 74 +/ 7); patients with moderate hyperhomocysteinemia tHCY 18 mumol/l for the women; > 20 mumol/l for the men; 21 patients, M/F = 18/3; mean age 74 +/- 7). RESULTS: As against the individuals with normal serum concentrations of HCY (Fig. 2; chi 2: p < 0.05), the patients with mild hyperhomocysteinemia (Odds ratio: OR = 1.48) and, above all, patients with moderate hyperhomocysteinemia (OR = 4.6) were found to have a greater prevalence of carotid lesions at high thromboembolic risk. No significant differences within the three groups were found with regard to distribution by age, gender and the prevalence of the more common cardiovascular risk factors (smoking, diabetes mellitus, arterial hypertension, hyperdyslipidemia). CONCLUSIONS: Hyperhomocysteinemia is associated with highly severe carotid lesions at higher risk for cerebrovascular events. PMID- 10670255 TI - [Carotid lesions and vascular risk factors]. AB - BACKGROUND AND AIMS: Recent studies have underlined a significant incidence of peripheral arterial occlusive disease (PAOD) of the lower limbs in the general population which is often wrongly diagnosed. The "classic" risk factors--like dyslipidemia--are not always present in significant percentages in patients suffering from PAOD of the lower limbs. The aim of this study was to evaluate the incidence of the most common vascular risk factors (smoking, hypertension, hyperglycemia, dyslipidemia) in patients suffering from stenosing lesions of the extracranial carotid axes, comparing the data with similar findings in lower limbs. Moreover, the authors evaluated the association between these risk factors, carotid atheromatous lesions and ischemic cardiomyopathy (CI). METHODS: A retrospective study was performed to evaluate the data from 1643 patients examined consecutively. A total of 636 (age > 40, carotid stenosis > 40%, presence of risk factors) were considered eligible. RESULTS: The results showed that, contrary to the findings in patients suffering from PAOD, diabetes was not among the most frequently associated risk factors, whereas a relatively large number of patients had a history of smoking, including both smokers and ex smokers. CONCLUSIONS: The difference in the most frequent risk factors identified for PAOD and carotid lesions suggests different etiopathogenetic mechanisms for the two districts. PMID- 10670256 TI - Hemorheology and tissue oxygenation in hypertensives with lipoidoproteinosis and peripheral occlusive arterial disease (POAD) treated with sulodexide and pravastatine and evaluated with laser assisted optical rotational red cell analyzer (LORCA) and transcutaneous oxymetry. AB - BACKGROUND: During arterial hypertension it is often possible to find other factors like lipoidoproteinosis and peripheral arterial disease (POAD), which can accentuate blood rheological abnormalities in hypertensive subjects. A group of hypertensives with lipoidoproteinosis (LP) and POAD were therefore examined to evaluate the relationship between these factors and blood rheological disorders and, if possible, to correct it. METHODS: We studied a group of 27 hypertensives with LP and POAD (15 males and 12 females in menopause for at least 1 year, aged 48 +/- 4 years), with WHO stage I hypertension, obesity (BMI = 30 +/- 2), stage II type "a" POAD, class 2 type "b" lipoidoproteinosis (acc. to Fredrick-son's classification) and hyperfibrinogenemia. All patients received oral medication with 500 lipidic units (ULS) sulodexide a day, 20 mg pravastatin o.d. orally, and were put on a low-salt and low-calorie diet (1400 kcal/day) during a follow-up of 60 days. Blood rheology status was evaluated before and after treatment (red blood cell--RBC--deformability and aggregability) using a new computerized instrument, which uses laser rays: the laser assisted optical rotational red cell analyzer (LORCA) (acc. to Hardeman) and RBC deformability using optical microscopy under immersion (acc. to Zipursky and Forconi). Transcutaneous oxymetry was also used to evaluate tissue oxygenation. RESULTS AND CONCLUSIONS: At the end of the study a significant improvement (p < 0.01) was noted in the blood rheological patterns of peripheral perfusion and tissue oxygenation. This underlined the positive influence of sulodexide with pravastatin in improving hemorheological patterns and modulating hypercholesterolemia and hyperfibrogenemia in hypertensives with POAD II "a" and LP 2 "b" and blood rheology disorders. PMID- 10670257 TI - [Formulating a non-fat diet to induce essential fatty acid deficiency in rats]. AB - OBJECTIVE: To describe the manufacture of an experimental diet to cause essential fatty acids deficiencies in rats. MATERIAL AND METHODS: Rats. We used Wistar rats that were given a diet consisting of skimmed milk, starch, and dextrino maltose. The fatty acids were measured by means of gas chromatography. RESULTS: The prepared diet is considerably cheaper than laboratory animal feed and it is effective in causing a fatty acid deficiency. PMID- 10670258 TI - [Descriptive analysis of the nutritional support in a polyvalent intensive care unit. Complications of enteral nutrition]. AB - OBJECTIVE: Description of the nutritional support in an intensive care unit. REFERENCE POPULATION: Patients hospitalized in our ICU over a period of 48 months (October 1994-September 1998). INTERVENTIONS: The study was carried out by means of a review of the two data bases generated, one by using the clinical history management program, and the other by using the artificial nutrition program. RESULTS: Nutritional support is used in 31% of the non-coronary patients, predominantly medical (61%), and followed by surgical (29%) and trauma (9%) cases. These patients presented an APACHE (17.7 +/- 15), a hospitalization (15.8 +/- 14.9) and a mortality (26%) that was greater than that in non-coronary patients who did not require the nutritional support. The delay in starting the nutritional support is 2.8 +/- 1.9 days. In decreasing order, the nutritional support is most used in medical (42%), trauma (37%) and surgical (18%) patients. The access route is similar, enteral in 55% of the cases, with a predominance of medical patients, and parenteral in 45% of the cases, with a predominance of surgical patients. In 100 patients with a nutritional support in excess of 10 days, it was found that 87% at some time were given this enterally. In this group we studied the gastrointestinal complications, finding these in 61% of these patients, with the most frequent complication being an increase in the gastric residue (44%). Diarrhea was found in 14% and broncho-aspiration in 3.4%. The enteral route as the initial access failed in 25% of these cases, thus requiring parenteral nutrition. CONCLUSIONS: In our unit we used nutritional support in 31% of the non coronary patients, and these presented a greater severity, longer hospitalization, and higher mortality than those patients who did not require this. The beginning of the nutritional support is relatively early. The gastrointestinal complications derived from enteral nutrition are very common, with a predominance of gastric retention. In 25% of the critical patients who begin enteral nutrition, this fails, and thus they require parenteral nutrition. PMID- 10670259 TI - [The usual diet of a group of adolescents from Valencia]. AB - Adolescents are considered a high nutritional risk group because their nutritional needs are increased with respect to other age groups, and because this period of life coincides with changes in life style that affect, often negatively, their eating habits. Our overall goal is to study the usual eating pattern of Valencian adolescents together with their drink and tobacco consumption, but the first stage focuses on setting up, validating and correcting methods to be applied. The present study was therefore carried out in a sample composed of 64 adolescents, ranging from 16 to 20 years of age. A self administered survey developed in our Department was used to explore their food preferences, eating habits, smoking habits and alcohol and coffee intake. The following results were obtained: The number of daily meals was of 3.7 +/- 0.9. The 91% takes the breakfast daily (milk with cereals or sweet rolls) and the majority of the students eat a second mid-morning breakfast. Lunch is of the traditional type, consisting of two courses. The first is rice or pasta, followed by meat, fish or eggs usually accompanied by a side dish or salad. Fresh fruit is the dessert eaten, almost daily by the majority of the sample. The most usual drink is water. The 70% of the sample have one supplementary afternoon intake "merienda". All of the surveyed people like fruits, pasta and chicken meat. While, liver and legumes are disliked by the majority. The intakes of soft drinks, snacks, alcohol, coffee and tobacco are moderate, being, all these products mainly consumed during the weekend. The mean diet offers an excess of proteins and saturated fat, while complex carbohydrates and dietetic fiber are scarce. Nutritional intakes of iron, magnesium and zinc in girls, and magnesium, folates and vitamin A in boys are estimated insufficient to fulfil their needs. PMID- 10670260 TI - [Methods for registering food intake in laboratory animals]. AB - Registering the ingestive activity of laboratory animals can be carried out by using electronic scales connected to personal computers, thus gathering data files that allow calculation of the daily ingestion standards. The ingestion parameters most often used in the standard analysis include the number, size, and duration of the meals, the speed of ingestion, and intervals between feedings, and the hunger and satiation indexes. It is necessary to know a minimum interval between meals to distinguish between true meals and pauses within one and the same meal. PMID- 10670261 TI - [A patient with visceromegaly: risk factor for performing percutaneous endoscopic gastrostomy. A clinical case]. AB - Percutaneous endoscopic gastrostomy is an easy and safe technique to provide an enteral access for patients needing long-term enteral nutrition. Minor complications may occur in 9% to 13% of patients. Life-threatening complications appear in 1-3% of cases. Perforation of a hollow viscus leading to peritonitis is a rare condition; hepatic perforation after placing a percutaneous endoscopic gastrostomy (PEG) has not been reported previously. In patients with massive visceromegaly an abdominal ultrasound may help in localizing the place of punction avoiding surrounding organs. PMID- 10670262 TI - [Anesthetic status monitoring: reality or pending matter?]. PMID- 10670263 TI - [Comparison of sevoflurane and propofol in the maintenance of and recovery from anesthesia]. AB - OBJECTIVE: To compare the recovery of patients after anesthesia with sevoflurane or propofol during open urological surgery or lumbar column surgery of intermediate duration. PATIENTS AND METHODS: Thirty-six ASA I, II or II patients were enrolled prospectively and randomly assigned to two groups to receive either sevoflurane (n = 19) or proporol (n = 17). Anesthetic induction was accomplished with thiopental, fentanil and vecuronium. During anesthetic maintenance a mixture of 60% nitrous oxide in oxygen plus the drug under study was adjusted to keep blood pressure and/or heart rate within +/- 20% of baseline. After surgery we recorded time until eye opening, spontaneous breathing, extubation, orientation, and identification of parts of the body. Side effects were likewise recorded. In the postanesthetic recovery ward patient condition was assessed using the Aldrete scale, the Newman-Trieger test and a visual analog scale for postoperative pain. Consumption of analgesic during the first 24 h after surgery was monitored. RESULTS: No significant differences were found in demographic data; duration of anesthesia; anesthetic doses; or time until spontaneous breathing, extubation, orientation or identification of parts of the body. Only time until eye opening was shorter in the sevoflurane group than in the propofol group (6.9 +/- 3.3 vs 11.5 +/- 4.8 min; p < 0.05). No differences were recorded on scales reflecting intermediate-term recovery. Analgesic consumption and the incidence of side effects were similar in both groups. CONCLUSIONS: Sevoflurane and propofol are comparable for anesthetic maintenance in urological and neurological procedures of intermediate duration. PMID- 10670264 TI - [Efficacy of 0.1 mg of subarachnoid morphine combined with bupivacaine on postoperative analgesia in total hip arthroplasty]. AB - OBJECTIVES: To analyze the analgesic efficacy, safety and side effects of subarachnoid morphine (0.1 mg) with bupivacaine in patients undergoing total hip arthroplasty. PATIENTS AND METHODS: Thirty patients scheduled for total hip replacement under spinal anesthesia with bupivacaine were randomly assigned to two groups according to whether local anesthetic with 0.1 mg subarachnoid morphine was also provided or not (group M [n = 15] and group S n = 15[, respectively). Top-up analgesia with morphine was available through a patient controlled device. Postoperative pain was assessed on a visual analogue scale (VAS) and consumption of intravenous morphine in the first 48 hours after surgery was recorded. RESULTS: VAS scores (mean +/- SD) were significantly lower in the first six hours in group M, but no differences between the two groups were observed thereafter. Total consumption of morphine at 48 hours was much lower in group M (6.80 +/- 7.74 mg) than in group S (31.38 +/- 13.17 mg). The incidence of nausea was high in both groups (46%). Slight pruritus affected 26.6% of patients in group M. Urinary retention necessitating temporary placement of a catheter was observed only in group M, where the incidence was 35.7%. No cases of respiratory depression occurred. Drowsiness was observed in 26.6% of patients in group S in comparison with 6.6% in group M. CONCLUSIONS: Combining 0.1 mg morphine and bupivacaine for total spinal anesthesia during hip arthroplasty significantly decreased the consumption of intravenous morphine during the first 48 hours after surgery. No respiratory depression occurred and the only side effects were urinary retention and mild pruritus and drowsiness. PMID- 10670265 TI - [Changes in oxygen saturation in the jugular bulb during cardiac surgery]. AB - OBJECTIVE: Heart surgery with cardiopulmonary bypass (CPB) leads to changes in supply and consumption of cerebral oxygen (DO2 and VO2C). Monitoring jugular bulb oxygen saturation (SjO2) detects changes in the DO2C/VO2C ratio that occur in patients undergoing heart surgery. The objective of this study was to determine the evolution of SjO2, of the arteriovenous difference of cerebral oxygen and of cerebral oxygen extraction, as well as the possible relation between those variables and changes in mean arterial pressure, hemoglobin counts and temperature in patients undergoing heart surgery with CPB. PATIENTS AND METHOD: A prospective study carried out in 31 patients who underwent coronary valve surgery. To monitor SjO2, each patient's internal jugular vein was cannulated with an oximetric catheter in a retrograde direction to monitor SjO2. RESULTS: Baseline SjO2 (68 +/- 7.4%), obtained after anesthetic induction, was similar to SjO2 before (65 +/- 6%) and after (67 +/- 8.2%) CPB. However, SjO2 upon starting CPB (60 +/- 8.6%) and during rewarming (63 +/- 3%) were significantly lower than at baseline. SjO2 was significantly higher during hypothermic bypass (78 +/- 5%) than at baseline. SjO2 ranged from a low of 60 +/- 8% as CPB was initiated to a high of 78 +/- 5% during hypothermic CPB. Mean arterial pressure was significantly lower at the start of bypass (44 +/- 6 mmHg) than anesthetic induction (83.5 +/- 13.1 mmHg) and the decrease correlated with a significant decrease in SjO2. Changes in mean arterial pressure were unrelated to significant changes in SjO2 at other moments, however. Nor was there a significant relation between changes in temperature or hemoglobin and the evolution of SjO2. At least one episode of SjO2 desaturation (= 50%) occurred in 29% of the patients, with the lowest values being recorded at the start of CPB and during rewarming. CONCLUSIONS: The greatest risk of cerebral oxygen imbalance between supply and demand occurs at the start of CPB and during rewarming, as shown by decreases in SjO2 levels below baseline at those times. PMID- 10670266 TI - [Transfusion needs during intraoperative and immediate postoperative periods in arthroplasty of the hip and knee]. AB - OBJECTIVES: To determine the factors associated with immediate perioperative transfusion requirements of hip or knee arthroplasty patients who have not been enrolled in a blood salvage program. PATIENTS AND METHODS: This prospective study collected demographic (age, sex, weight, height, etc.), physiological (hemoglobin levels, coagulation times, preoperative platelet counts, etc.), clinical history and anesthetic and surgical data (type of anesthesia, surgical diagnosis, duration of procedure) in 112 patients undergoing orthopedic surgery: 19 cases of primary knee arthroplasty, 77 cases of hip arthroplasty and 16 replacements of hip arthroplasty. Logistic regression analysis of the aforementioned variables was performed to search for factors related to transfusional needs during and after hip arthroplasty or after knee arthroplasty, which was performed with a tourniquet applied to render intraoperative transfusion unnecessary. RESULTS: The variables that increased the risk of transfusion during surgery were duration of procedure exceeding 120 min (OR 15.24; p = 0.01) and loss of over 500 ml of blood during surgery (OR 11.4; p = 0.02). The variables associated with perioperative transfusion were loss of over 500 ml in the postanesthetic recovery room (OR 12.6; p < 0.0001), hypotensive episodes during recovery (OR 11.7; p = 0.0001), prosthetic replacement (OR 6.33; p = 0.005), height < 160 cm (OR 5.03; p = 0.02), preoperative hemoglobin level < 13.5 g/dl (OR 4.97; p = 0.02), and surgery for reasons other than osteoarthritis (arthritis, pathological fractures, etc.) (OR 4.60; p = 0.04). Variables associated with transfusion of over two units of packed red cells were a history of neoplastic disease unrelated to arthroplasty (OR 378.67; p = 0.005), prosthetic replacement (OR 49.71; p = 0.009), diabetes (OR 36.49; p = 0.02) and a hypotensive event while in the postanesthetic recovery room (OR 29.12; p = 0.02). CONCLUSION: These results suggest that certain modifiable factors increase the risk of blood transfusion in knee and hip arthroplasty. Specifically, they are duration of surgery, intra- and postoperative bleeding, preoperative hemoglobin level and instances of perioperative hypotension. Other factors outside our control are height or patient clinical history. PMID- 10670267 TI - [Ropivacaine]. AB - This paper describes the pharmacological, pharmacokinetic and pharmacodynamic features of ropivacaine on both an experimental and clinical level. Ropivacaine is an amide-type local anesthetic whose chemical structure is related to that of mepivacaine and bupivacaine and whose duration of effect falls between the two. Ropivacaine is a less potent effector of motor blockade than bupivacaine, and its toxic effects on the central nervous system and myocardial tissue is likewise less. Ropivacaine has been employed for epidural anesthesia and analgesia (including through a caudal approach), for peripheral motor blockade, for local infiltration (in gel form), and for intravenous regional anesthesia in ophthalmologic surgery involving peri- and retrobulbar blockade. Subarachnoid use has not yet been accepted in Spain, although phase IV clinical trials of this application have begun. Concentrations of 2 mg/ml (0.2%) and 7.5 mg/ml (0.75%) and 10 mg/ml (1%) are available in Spain. PMID- 10670268 TI - [Headache caused by endocranial hypotension secondary to sacrococcygeal trauma in a patient with a congenital arachnoid cyst]. AB - Headache caused by intracranial hypotension after sacrococcygeal trauma, is a rare syndrome with very similar symptoms and physiopathology to post dural puncture headache. In both situations, cerebrospinal fluid (CSF) leaks through a dural tear, leading to a decrease in its pressure. A 13 year old patient presented a frontal and occipital positional headache, after undergoing sacrococcygeal trauma. In magnetic resonance images, the presence of a spinal arachnoid diverticulum (cyst) and CSF leak were confirmed. After the establishment of conservative medical treatment, complete clinical remission was observed. The diagnosis of intracranial hypotension headache syndrome is mainly clinical, once other possible headache etiologies have been discarded. Magnetic resonance imaging can be used to detect small CSF leaks, and in this case, magnetic resonance imaging clearly showed the existence of an arachnoid cyst. The presence of some meningeal diverticulums such as arachnoid cyst, constitute a risk to undergo this syndrome, due to the possibility of its rupture by minor traumatisms. The initial treatment should be conservative, as the dural tear seals spontaneously and the liquid is reabsorbed. PMID- 10670269 TI - [Remifentanil and sevoflurane in laparoscopic bariatric surgery]. PMID- 10670270 TI - [Paradoxical air embolism during catheterization of the right subclavian vein]. PMID- 10670271 TI - [Pre-eclampsia and treatment with urapidil]. PMID- 10670272 TI - [Difficult intubation in a case of ankylopoeitica spondylitis]. PMID- 10670273 TI - [Complete obstruction of a urinary catheter caused by urate deposits associated with a continuous perfusion of propofol (ivofol)]. PMID- 10670274 TI - [Anterior bridges with the IPS-Empress-2 System after alveolar ridge augmentation. A case report]. AB - The success of a prosthesis is judged according to optimal function, good chewing comfort, adequate phonetics and white and pink esthetics. The aim of a treatment is to approach the perfection of nature. For anterior bridgework, the all-ceramic System IPS Empress 2 offers light transmission and reflection comparable to that of natural teeth, provided that the pink esthetics are optimised in the preprosthetic phase. The provision of an anterior bridge in the IPS Empress 2 system is presented here in the form of a case report. After extraction of the anterior teeth, a ridge augmentation including preparation of the pontic bed was carried out. The type of post and core, preparation and cementation are important parameters for the success of all-ceramic restorations. PMID- 10670275 TI - [The pregnant patient in dental care. Survey results and therapeutic guidelines]. AB - In a telephone survey using a standardized questionnaire, 78 resident dentists in Germany, Switzerland and Austria were interviewed with respect to several aspects of the dental treatment of pregnant women. Only 58% of the interviewees decided clearly in favour of local anaesthetics, 59% supported the use of analgesics, 70% a possible antibiotic therapy and 33% a radiological examination during pregnancy. In addition, according to references in the specialist literature guidelines for the dental treatment, drug therapy and radiological diagnosis of pregnant women are presented. The local anaesthetics should have a high plasma protein bonding (articain, bupivacain, etidocain) and a minimum adrenaline concentration. Paracetamol is the analgesic of choice. If an antibiotic treatment is required, penicillin, cephalosporin and erythromycin are recommended. In particular during the first three-month period, radiological examinations should be restricted to the absolute minimum and performed only if no reasonable alternative is available, even though the radiological burden on the foetus falls 500,000 times short of the limit value of 50 mgray (5 rad) in the case of a microradiogram, and 50,000 times short of the limit value in the case of an orthopantomogram. PMID- 10670277 TI - [Antibiotic resistance: current data from around the world and Tunisia]. PMID- 10670276 TI - [Consumption of antibiotics in Tunisia]. PMID- 10670278 TI - [Methods for estimating the cost of nosocomial infections]. PMID- 10670279 TI - [Markers of viral hepatitis in blood donors. Study apropos of 300 donors]. AB - In order to locate the rate of positivity of the different markers of hepatitis that can be transmitted through blood (Ag HBs, anti-HBc Ab, anti-HCV Ab and transaminases (ALAT)), we have conducted a study over 300 blood donors and who are sale, equally shared between the parts of age and of sex. The ALAT were superior to the normalcy in 10.66% of the cases, the male sex being 3 times more affected than that of the female. The Ag HBs was positive in 4.66% of the cases. The male sex is twice as affected as the female sex. One of three persons who donated blood (37.33%) had anti-HBc Ab. Finally none of the 300 blood donors has the anti-HCV Ab. There's a very weak recovering between the presence of Ag HBs and of ALAT superior to the normalcy, a weak recovering between Ag HBs and anti HBc Ab and a certain recovering between ALAT superior to the normalcy and the presence of anti-HBc Ab. There is no recovering between the anti-HCV Ab and the indirect markers. Eventually, the systematization of all these tests over each donation engenders the elimination of more than 2/5 of the collected products. PMID- 10670280 TI - [Diagnostic and therapeutic aspects of lymphatic tuberculosis: apropos of 42 cases]. AB - We report a retrospective study of 42 cases of lymph node tuberculosis. We noted symptoms of tuberculosis impregnation in 92%, cervical localization in 71%, positive tuberculin intra-dermo-reaction in 77%, and accelerated erythrocyte sedimentation rate in 73% of the cases. Koch bacillus was detected in expectoration, urine or gastric liquid at the rate of 11% of the cases. Lymph node function was suggestive in 4 out of 12 patients, showing giant cells with or without caseum. Lymph node biopsy, performed in 32 patients, was contributive in 94% of them. Another tuberculous localization was found in 14 cases mainly pulmonary (8 cases). PMID- 10670281 TI - [Detection and molecular typing of human papillomaviruses: prevalence of cervical infection in the Tunisian central region]. AB - There are compelling molecular and epidemiological data which indicate that infection with certain genital human papillomaviruses (HPVs), such as HPV 16 and HPV 18, has a critical role in initial changes that lead to cervical and probably other anogenital cancers. These observations prompted us to investigate the prevalence of cervical infection with genital human papillomaviruses in Tunisia. We used the polymerase chain reaction (PCR) to detect and type HPV DNA. The prevalence of HPV infection in a population of 106 Tunisian women recruited at the Offices Nationaux de la Famille et de Population (ONFP) was 13.6%. Molecular HPV typing indicated a high prevalence of HPV at high oncogenic risk; Our results indicate that the infection with genital human papillomaviruses is frequent in the Tunisian population. PMID- 10670282 TI - [Cavopulmonary shunts: apropos of 40 observations]. AB - Fourty patients with univentricular heart, underwent a cavopulmonary shunt procedure. The majority have an excellent hemodynamic status with ventricular end diastolic pressure > 12 mmHg and a mean pulmonary artery pressure > 15 mmHg. However, 11 patients have ventricular dysfunction, 9 have an incompetent systemic atrio-ventricular valve and 6 have mean pulmonary artery pressure > 15 mmHg. The pulmonary arteries were of a good size in all cases with a Nakata index > 100 mm2/m2. Cavopulmonary connections are satisfactory palliative procedures in the treatment of univentricular cardiac disease. PMID- 10670283 TI - [Spontaneous contrast of the left atrium in mitral valve stenosis]. AB - The left atrial spontaneous contrast is uncommon finding during transthoracic echocardiography. Transesophageal echocardiography provides superior imaging of the left atrium and left spontaneous contrast has been detected more frequently by this technique in rheumatic mitral stenosis. In order to analyze the significance of spontaneous contrast, we have studied 100 patients with mitral stenosis. Trans thoracic and transesophageal echocardiography were performed in all patients. Left atrial spontaneous echo contrast was detected in 60 patients (group A) and was absent in 40 patients (group B) The mean of mitral valvular area was 0.9 10.2 mm in group A and 48 9.5 mm in group B (p < 0.001). Ther was a high incidence of atrial fibrillation in group A; 63% VS 12% in group B (p < 0.001). A systemic embolization was noted in 20% of patients in group A; No one of group B has presented this complication (p < 0.001). PMID- 10670284 TI - [Prognosis of intraventricular septal rupture in myocardial infarct: apropos of 13 cases]. AB - The authors report the experience about 13 cases of postinfarction ventricular septal rupture. Patients (9 men and 4 women) aged 60 to 76 years average 68 years. The site of necrosis was the anterior wall in 8 cases and the posterior wall in 5 cases. The diagnosis of septal rupture was confirmed by echocardiography and/or angiography. 6 patients died after averages: 7 days. Surgical intervention was performed in 7 cases after a mean of 51 days following the date of the myocardial infarction after an intensive care. We emphasize to demonstrate the helpful of intensive care and the role of early surgery for a good prognosis. PMID- 10670286 TI - [Postpartum myocardial infarct (apropos of a case)]. AB - We report the case of a 26 years old woman, who was hospitalized for an acute anterior myocardial infarction, which happened 17 days after delivery. The culprit lesion was a coronary dissection observed on the coronary angiogram which was performed on the 7th day. This dissection cicatrised spontaneously at the control coronarography performed 3 months later. Conventional medical treatment seems to be sufficient, although the use of thrombolysis in our case was safe and successful. PMID- 10670285 TI - [Mixed form of total anomalous pulmonary venous connection: apropos of a case]. AB - As opposed to partial anomalous of pulmonary venous connection, it is frequent and benigns, the total anomalous of pulmonary venous connection is extremely rare and more serious. The anomalous is severe because all pulmonary venous connection, instead of left heart it go to the right heart. The age of diagnosis is closely tied up anatomics characteristics, so various clinical cases are present. The TAPVC of new born is a surgical emergency, especially where it's block up and release++ cardiorespiratory distress syndrome secondary to OAP. For great children, the total anomlous of pulmonary venous connection can be assumed to a case of atrial septal defect. This study intend to clear up this clinical and anatomical polymorphism and to report an exceptional-form of this congenital anomaly which is mixed total anomalous of pulmonary venous connection. PMID- 10670287 TI - [Colonic tuberculosis: an exceptional cause of a massive surgical hemorrhage apropos of a case and review of the literature]. AB - Colonic tuberculosis rarely causes massive bleeding, as a matter of fact less than twenty cases have been reported in medical literature. Resorting to surgery in this context is even more exceptional. We have not found any case operated on in the literature we have consulted. Endoscopy together with histological study of biopsies is the best exam for the diagnosis of this disease. We report the case of a 37 year old male, coronarian, carrying a caecal lesion evocative of tuberculosis at the endoscopic exam, who had an emergency right hemicolectomy for massive bleeding caused by caecal tuberculosis. The surgical indication was dictated by the abundance and the persistence of the bleeding. The post-operative course was uneventful. PMID- 10670288 TI - [Effect of fasting on glycemia and proteinemia in young and adult rats]. AB - The aim of Our work has consisted of studying the effects of a severe three-day fast on the evolution of the body weight, on glycemia and on proteinemia among the young and mature male rats. The decrease in body weight which is more marked among young rats after starvation shows that the loss of body mass due to fasting decreases with the age of the rats. The physiological perturbations of nutritive constants that have been recorded during this period of fast show that the mature rat first attacks its non-protein stocks or glucides, which leads to a decrease in its glycemia and in its lipids; whereas the young rat uses its proteins after only three days of starvation because of its small stock of lipids. PMID- 10670289 TI - Healthcare professionals must learn to say sorry. PMID- 10670290 TI - Can doctors' work really be expert nursing? PMID- 10670291 TI - Case 7: breach of confidentiality. Psychiatric nurse talks about a client to a newspaper and radio. PMID- 10670292 TI - Evaluating the Pegasus Trinova: a data hierarchy approach. AB - Understanding the efficacy of patient support surfaces is essential if pressure sore management is to be both efficient and effective. However, laboratory and clinical studies in this area are fraught with well recognized problems. This investigation reports a combination of laboratory, randomized controlled trial (efficacy data) and measures of effectiveness to illustrate the beneficial role of a new dynamic integrated mattress and seat cushion system: the Pegasus Trinova. Successful prevention of sores among a vulnerable patient population, along with positive comments regarding the system's comfort and 'user friendliness' are supported by laboratory measures of interface pressure to provide a hierarchy of data. Such an approach may present one solution to the lack of timeliness of most mattress clinical trials, thus allowing decisions regarding new support surfaces to be made upon the basis of evidence, not on anecdote or solely upon marketing claims. PMID- 10670293 TI - A practical approach to dressing wounds in difficult positions. AB - Despite the availability of an increasing number of innovative dressing products, many patients' wounds remain a challenge to dress in practice. However, the creative use of widely available products and other resources can help to address some of the difficulties that are regularly encountered. If different sizes of dressing (to allow for cutting and shaping) or more innovatively shaped products (which may be more expensive per item) are used, this can result in a better dressing fit and therefore increase product wear time which is both clinically effective and cost-effective. When a good solution is found for dealing with certain wound types then great benefit can be gained from publishing the information to prevent others from struggling to overcome the same difficulties. PMID- 10670294 TI - Family carers. 4: Designing services to support family carers in Sweden. AB - This, the last of four articles focusing on family caregiving in Sweden, considers ways in which services can be designed to better support and complement care provided by the family. Drawing on the results presented in the previous three articles (Vol 8(9): 582-8; Vol 8(10): 647-52; Vol 8(11): 735-40) the need to provide flexible and responsive services will be highlighted. While acknowledging the importance of services which help with the physical aspects of caring, emphasis will be placed on the need to develop an expanded range of interventions which recognize and respond to the social and emotional impact of caring. PMID- 10670295 TI - Confidentiality. 4: Patient confidentiality and the courts. PMID- 10670296 TI - Autonomy or non-compliance in adolescent diabetes? AB - Many physical, psychological, social and cognitive changes take place in adolescence. Emerging personal values and beliefs, an acute awareness of body image and a desire for peer conformity and increasing independence can make the transition to adulthood troublesome. For teenagers with diabetes, usual adolescent needs and concerns are complicated by the demands of a complex treatment regimen. This can result in non-adherence to the recommended programme of care and, consequently, poor glycaemic control, which increases the risks for diabetes-related complications in later life. This situation poses a major problem for healthcare professionals. Teenagers, however, are frequently preoccupied with the present, and are unable to perceive, or will ignore, their vulnerability to long-term health problems. This article discusses these difficult issues and, taking into account the increasing need for autonomy and independence in adolescence, makes recommendations for nursing practice. PMID- 10670297 TI - The benefits of preparing children and parents for day surgery. AB - Hospital admission for children inevitably provokes feelings of anxiety for both parent and child. The development of day surgery has in some respects eased many of these anxieties; however, without the support of both verbal and written information and the development of quality preparation from children's nurses anxiety will continue to exist. This article seeks to explore the benefits of preparing children and parents for day surgery using preadmission education, and suggests how improvements may be made to practice, education and research. PMID- 10670298 TI - Cervical cancer. 2: Colposcopy, treatment and patient education. AB - In the first article in this two-part series (Vol 8(11): 730-34) the issue of cervical screening and the cervical smear test was described and discussed. This article looks at colposcopy. When a woman receives an abnormal smear test result, she will often require further examination of the cervix in order to identify the degree and area of dyskaryosis. Colposcopy can often provoke feelings of fear and anxiety in women and the nurse must help to alleviate these fears and anxieties. Colposcopy and cervical intraepithelial neoplasia are discussed and the treatment (if any) is outlined. PMID- 10670299 TI - Cardiopulmonary resuscitation: are practitioners being realistic? AB - Cardiopulmonary resuscitation (CPR) is now established medical practice for all in-hospital cardiac arrests except where a specific 'do not resuscitate' (DNR) order is in place. This article explores many of the ethical and moral issues surrounding CPR and the use of DNR orders. It examines the success rate of in hospital CPR and raises the question of what constitutes outcome success by illustrating that at best only 15% of resuscitated patients survive to hospital discharge. The article proposes that both patients and healthcare professionals grossly overestimate the success of CPR and suggests that many elderly patients might choose not to be resuscitated if they were allowed to make an informed choice. It concludes by suggesting that further work needs to be undertaken with regard to early assessment of all in-hospital patients, combined with realistic and frank communication between healthcare professionals and patients if futile, undignified and costly deaths are to be avoided. PMID- 10670300 TI - EasiCath: an advanced alternative to indwelling urethral catheters. AB - This article looks at EasiCath, the single-use hydrophilic catheter manufactured by Coloplast Ltd. EasiCath is now widely used throughout the UK and Europe. Since the launch of EasiCath, Coloplast has continued to develop its lubricated catheters and with the recent inclusion of the 30 cm EasiCath on the Drug Tariff, Coloplast now provides the most extensive range of lubricated catheters in the UK. In recent trials looking at the cell count on the surface of hydrophilic catheters following catheterisation, EasiCath performed well, indicating that it is a safe choice of lubricated catheter (Biering-Sorensen et al, 1999). This, combined with the benefits of the precoated, polished eyelets and the potential cost saving to the GP (4.60 Pounds per box of 25) over a number of other catheters available, means that EasiCath has become the first choice of hydrophilic catheter for many patients. PMID- 10670301 TI - Healthcare risk management: 18. Clinical audit systems. PMID- 10670302 TI - Nurse anaesthetists should not be encouraged. PMID- 10670303 TI - Nutrition and hydration are not options but rights. PMID- 10670304 TI - Protecting people with disabilities from abuse. PMID- 10670305 TI - Case 8: misappropriation of ward funds. Ward manager who took money from the ward and a patient. PMID- 10670306 TI - Measuring the size of the leg ulcer problem in an acute trust. AB - The majority of patients with leg ulceration are cared for in the community. There is insufficient evidence of the numbers of these patients who are being cared for in acute hospitals. This study sought to identify the size of the problem in one acute trust by means of a point prevalence survey. From a study population of 931 patients, 17 had leg ulcers, giving a prevalence of 1.8%. Out of 52 wards and units, staff on 44 (84.6%) said that patients with leg ulcers were admitted on occasion. In the case of 19 (36.5%) wards and units this was as infrequently as one or two patients a year. The results of this survey have considerable implications for ensuring that all patients receive optimum care. PMID- 10670307 TI - Inflammatory bowel disease. 1: Aetiology and pathogenesis. AB - Inflammatory bowel disease, a chronic lifelong condition, affects between 15 and 30 people per 10,000 of the UK population. Despite the large number affected, there seems to be a paucity of information on this subject in the British nursing press. Hence, nurses have little or no understanding of the impact of this illness, and there is a danger that the distress and debilitation that inflammatory bowel disease can have on people's lives is underestimated. However, a healthcare professional, armed with the appropriate knowledge and attitude, can make a real difference to the quality of care that these patients receive. This article is the first in a series of three that shall review the aetiology, treatment and concept of patient participation in inflammatory bowel disease. The series aims to enhance nurses' understanding and stimulate further investigation into this chronic illness. PMID- 10670308 TI - Confidentiality. 5: The rights of transplant recipients and donors. PMID- 10670309 TI - Learning disabilities: supporting nurses in delivering primary care. AB - This article introduces an educational project supported by The Queen's Nursing Institute and sponsored by the Hertfordshire Nursing Trust. It reports on the initial results from a literature review and its implications for nursing. There is broad acknowledgement of a problem in the delivery of primary care nursing for people with a learning disability, but little consensus on how to move either medical or nursing practice forward. It was proposed, therefore, to systematically assess the learning needs of nurses delivering primary care and to develop an evaluated methodology for responding to those needs. A comprehensive literature review was undertaken. Three consistent themes emerged: knowledge and training needs; role definitions and liaison; and clinical issues. Implications for nursing, and the future development of the project are considered. PMID- 10670310 TI - Preventing rape and sexual assault of people with learning disabilities. AB - This article will explore the increased prevalence of rape and sexual assault among people with a learning disability and identify factors that exist which increase this client group's vulnerability to becoming a victim of sexual abuse. There is significant under-reporting of such incidents and very few cases end up in prosecution. Within the nursing profession, there is much more that can be done to help decrease the level of vulnerability of people with a learning disability and to work with other agencies to advocate for legal justice and therapeutic redress for individuals. PMID- 10670311 TI - Nocturia, nocturnal polyuria and secondary nocturnal voiding. AB - Getting up once to the toilet at night is normal. Getting up more than once may be abnormal. Potential causes include: nocturia, which is bladder related; nocturnal polyuria, which is cardiac in origin; and being awake for a variety of reasons that are not linked to the bladder. Bladder problems, therefore, are not always the reason for frequency in micturition at night and treatment will only be effective if the correct cause and/or causes are identified. The most useful investigation to help with diagnosis is a frequency volume chart linked to an assessment identifying causes of secondary nocturnal voiding and nocturnal polyuria. Getting up to the toilet at night will not always be successfully treated by anticholinergics. Charting is the key to diagnosis, appropriate interventions and successful outcomes. This article will focus on working definitions, prevalence, causes, investigations and treatment options for nocturia, nocturnal polyuria and secondary nocturnal voiding. PMID- 10670312 TI - Current recommendations for isolation practices in nursing. AB - Attitudes have changed drastically over the centuries towards people with infections and how to contain them. Only as we approach the end of the 20th century are we starting to base our practices on scientific evidence and not on ritual, although rational thought is still not found in many practices and confusion surrounds the terminology used. With the introduction of clinical governance, and the statutory duty of health organizations to provide a quality service for patients supported by evidence-based practice, this article discusses isolation practices. PMID- 10670313 TI - Ethics and nursing research. 1: Development, theories and principles. AB - This article, the first of two looking at nursing ethics and research, outlines the foundations and development of an ethical framework for nursing research. The two dominant theories of ethics--utilitarianism and deontology--are described as they relate to the rights of individuals undergoing the research. Each of these approaches has limitations and in some instances choosing the right action may be difficult. The guiding ethical standards of beneficence/non-maleficence, respect for human dignity, justice, informed consent and vulnerable subjects are reviewed for the reader as they relate to undertaking research. This knowledge will help nurses conduct, participate in, or use research that is based on ethically sound principles. The second article will explore and explain the relationship between these guiding principles and the elemental steps of the research process. PMID- 10670314 TI - Clinical nurses urgently need a career structure. PMID- 10670315 TI - Cultural competency in nursing. PMID- 10670316 TI - The skin's antioxidant systems. AB - The skin is supplied with an antioxidant system that includes enzymatic and nonenzymatic components. This complex system is the skin's first-line defense against free-radical attacks. Antioxidants are essential in protecting the epidermis from damage by free radicals generated both by environmental and endogenous factors. This role of free radicals in etiology of disease and how antioxidants are used to offset or prevent oxidative damage are discussed. PMID- 10670317 TI - Heparin-induced skin necrosis: nurses beware. AB - As the prophylactic use of heparin continues to increase, nurses must be aware that heparin use may cause heparin-induced skin necrosis--a rare but serious complication. Although even more severe complications may occur from heparin use, this discussion will focus on skin necrosis caused by subcutaneous heparin. Should heparin-induced skin necrosis develop, heparin therapy must be discontinued immediately. This case presentation illustrates one patient's reaction to this complication. PMID- 10670318 TI - What's your assessment? Nummular eczema with eczema craquele. PMID- 10670319 TI - Pain management for dermatologic laser surgery: a nursing perspective. AB - Pain after laser surgery is one of the most commonly experienced patient problems. It is a pervasive part of laser treatment and a problem with which nurses and physicians are expected to deal. The nurse's most important role is preoperative patient teaching. Prior to undergoing the prescribed treatment, patients must have a realistic understanding of the laser procedure, including the expected sensations associated with the surgery. Education will not only correct misconceptions and reduce anxiety, but will also allow patients to play an active role in their surgery and to optimize fully from pain-reducing techniques and strategies. PMID- 10670320 TI - Dermatology nurse certified--now what does that mean? AB - What is certification and what does it mean to professionalization of dermatology nursing? The certification process is explored through one personal experience. Some discussion of the Dermatology Nurses Association's role in professionalism is included. PMID- 10670321 TI - A guideline for high-quality patient care. PMID- 10670322 TI - Autoimmune bullous diseases: diagnosis and management. AB - Bullous diseases constitute a wide group of disorders in which blistering of the skin and/or mucous membranes occurs. Patients may develop other lesions in addition to blisters. It is important for dermatology nurses to understand the pathogenesis of these disorders, the means for making an accurate diagnosis, and systemic and local management of the diseases. PMID- 10670323 TI - The beginnings of dermatology: a brief review. AB - Dermatology emerged as a distinct medical specialty in the mid-1800s. The background that led to this development, and the contributions made to the specialty by pioneering physicians in England, France, the German-speaking countries, and the United States are discussed. PMID- 10670324 TI - A review of cutaneous disease in African-American patients. AB - It is often difficult to determine the role race may play in diagnosis, most notably in patients with deeply pigmented skin. Among the members of any population, there are a significant range of skin colors and hair types which makes generalizing impossible. Deep pigmentation can mask anticipated cutaneous reactions or predispose to severe postinflammatory hypopigmentation and hyperpigmentation. It is imperative that professionals in dermatology recognize reactions in patients with deeply pigmented skin in order to diagnose and treat effectively. PMID- 10670325 TI - What's your assessment? Streptococcal sepsis. PMID- 10670326 TI - Traumatic wound care. AB - The scope and importance of traumatic wound care, assessment, debridement, pre and postoperative management, and subsequent skin care during the course of treatment cannot be over-emphasized, and indeed, are the most important considerations for functional and cosmetic outcome. Care begins in the emergent phase and continues through acute and convalescent phases. Efforts are directed at methods and techniques which prevent infection, facilitate wound healing, promote comfort, and at the same time, maintain optimal function and minimize deformities. PMID- 10670327 TI - Lasers for vascular lesions. AB - Recent developments in laser technology have revolutionized the treatment of various cutaneous disorders. Lasers provide effective and safe treatment of many conditions for which previous therapy was either unavailable, ineffective, or unacceptable. Basic laser principles, laser safety, available laser systems for treatment vascular lesions, clinical applications, preoperative considerations, anesthesia, postoperative changes, side effects/complications, nursing measures, and patient education are described. PMID- 10670328 TI - A new treatment option for acne scars: allograft dermis. AB - Unsightly facial scars resulting from acne can be the source of painful emotional trauma. A new option for treating acne scars is described. It is pre-packaged dermal graft material that is easy to use, safe, and effective. A review of current treatment options and the steps for using this material are presented. PMID- 10670329 TI - Wound assessment and evaluation. Diabetic neuropathic ulcer. PMID- 10670330 TI - Patient education for adults with chronic eczema. AB - Seven patients with severe eczema participated in an individual education project. The aim of the education program was to maintain and improve health by providing knowledge in self-care treatment. Patients were provided with new knowledge of their disease and received sufficient information to enable them to apply self-care of their eczema. PMID- 10670331 TI - What's your assessment? The diagnosis is herpes simplex. PMID- 10670332 TI - Alternative therapies, should we discourage or educate? PMID- 10670333 TI - Managing atopic dermatitis. AB - "Itchy" rashes occurring in patients (usually an infant or child) with a personal and/or family history of itching, sneezing, and/or wheezing should be considered part of the spectrum of skin problems labeled "atopic dermatitis" (AD). Multiple factors influence AD, and effective management requires that none be overlooked. The complicated nature of treatment means parents and family members require repeated reinforcement and education, particularly in avoiding "triggers of itch;" and use of medications. When all the aspects of AD are adequately addressed, more than 90% of patients with AD can be effectively managed. PMID- 10670334 TI - What's your assessment? Epidermoid inclusion cyst. PMID- 10670335 TI - Biomedical ethics and the role of the nurse: a case study and discussion. AB - Ethical issues are becoming of increasing concern to dermatology nurses as they struggle to maintain their role as patient advocate in the midst of cost constraints in the health care system. A case study is presented of one such dilemma. Suggestions are given for keeping nursing principles and advocacy at the forefront of patient care. PMID- 10670336 TI - Wound assessment and evaluation. Unstageable pressure ulcer due to eschar formation. PMID- 10670337 TI - Update on acne therapy. AB - Current therapy of acne vulgaris is very effective. It consists of a combination of topical comedolytic agents, antibacterial agents, and combinations of both. The use of systemic therapy with antibiotics, isotretinoin, and hormones is necessary for cystic acne. The management of patients with the various combinations of topical and systemic medications is discussed. PMID- 10670338 TI - The soul of managed care. PMID- 10670339 TI - Patient compliance: an issue for nursing. PMID- 10670340 TI - Emergency room dermatology. AB - Dermatologic problems are seen very often in the urgent care setting. Common acute problems such as urticaria, pruritus, and less common but potentially devastating dermatologic diseases will be discussed in this review. Emphasis will be placed on an efficient and cost-effective initial approach to diagnosis and treatment. PMID- 10670341 TI - Development of dermatology nursing certification. AB - In February 1998 the first Dermatology Nursing Certification exam was administered to registered nurses. This added credential beyond licensure serves as a demonstration that Dermatology Nurse Certified (DNC) nurses have acquired a core body of specialized knowledge and adhere to specialized nursing standards. In addition to providing a sense of professional achievement, certification can serve other purposes: peer and collegial respect, documentation of advanced knowledge and skills, increased earning power, increased job satisfaction, and in the future, third-party reimbursement. PMID- 10670342 TI - What's your assessment? Impetigo. PMID- 10670343 TI - Alopecia areata: identification and current treatment approaches. AB - Alopecia areata is a disease which occurs in 1.7% of the population (Safavi et al., 1995), often with devastating effects to patients and their families. In the past, this condition has been misunderstood and treated inadequately. New treatment modalities and support systems are offering hope to patients with alopecia areata. PMID- 10670344 TI - Wound assessment and evaluation. Venous stasis ulceration. PMID- 10670345 TI - Lymphedema awareness. PMID- 10670346 TI - Competency in the 21st century. PMID- 10670347 TI - Health care needs better towers. PMID- 10670348 TI - The relationship between skin cancer knowledge and preventive behaviors used by parents. PMID- 10670349 TI - Brown recluse spider bites. AB - The bite of the brown recluse spider (BRS) is the most severe arthropod cause of necrotic skin lesions. The severity of cutaneous injury due to BRS bites vary from mild erythema to severe necrosis. Rarely, severe systemic reactions occur. Diagnosis may be difficult, since the victim may not feel the bite or see the spider. Treatment must be individually tailored. Proper treatment, reassurance, and rapid followup are helpful in reducing the cutaneous and psychologic discomfort associated with BRS bites. PMID- 10670350 TI - Wound assessment and evaluation. Bullous pemphigoid. PMID- 10670351 TI - What's your assessment? Fixed drug eruption. PMID- 10670352 TI - Study habits and test-taking tips. AB - Everyone experiences test anxiety to some degree. Helpful suggestions about how to reduce test anxiety, how to study for tests, and how to develop good study habits can reduce this anxiety. Studying is different for different types of tests. It is important to learn how to study for multiple choice, true/false, short answer, and essay tests. These skills are also helpful when preparing for a certification examination such as the Dermatology Nurses Certification Exam. Use these tips and techniques to become better prepared for tests, and the suggested methods for reducing anxiety prior to and during tests. PMID- 10670353 TI - Achieving excellence in health care. PMID- 10670354 TI - What's wrong with this picture? PMID- 10670355 TI - Epidermolysis bullosa: a nursing perspective. AB - Caring for patients affected with epidermolysis bullosa (EB) can be one of the biggest challenges a dermatology nurse can face. Being familiar with EB and its associated complications is of utmost importance in caring for those individuals, otherwise patient care can be compromised. To provide the quality care that these individuals need, education is required for those affected and their family members as well as for health care providers involved in their care. PMID- 10670356 TI - The biopsychosocial burden of genital herpes: evidence-based and other approaches to care. AB - Genital herpes, an incurable disease, is increasing in epidemic proportions in the United States (Eng & Butler, 1997). Updated information on the epidemiology and psychosocial aspects of genital herpes is necessary to understand the biopsychosocial burden of living with the disease. Data from both qualitative and quantitative research are used to support evidence-based and other approaches to care. PMID- 10670357 TI - What's your assessment? Bowen's disease. PMID- 10670358 TI - The diagnosis of skin disease. AB - While there are certain common features in taking a history and doing a physical examination, every health care specialty has its own approach to gathering information. Dermatology is no exception. The purpose of this article is to outline in some detail the history taking and the physical examination commonly used to make a dermatologic diagnosis. PMID- 10670360 TI - Wound assessment and evaluation. Brown recluse spider bite. PMID- 10670359 TI - Patient education in the long-term management of atopic dermatitis. AB - Among the various forms of eczema, atopic dermatitis is the most common, distinctive, and difficult to manage. Educating patients about avoiding flare factors and proper use of moisturizers along with rational use of topical corticosteroids is crucial to effective long-term management. Openly reviewing facts and fallacies about allergies can help keep the focus on necessary skin care while dealing with patients' allergy concerns. The team approach of physician and nurse helps clarify confusing issues and maximizes patients' ability to effectively treat a chronic but controllable condition. PMID- 10670361 TI - Nursing problems. A study. PMID- 10670362 TI - Nurses and fight against tuberculosis--II. PMID- 10670363 TI - The Fifth Central Pay Commission. Salaries and allowances of nursing personnel. PMID- 10670364 TI - Nursing care of alcoholic liver disease. PMID- 10670365 TI - A survey on yellow oleander poisoning. PMID- 10670366 TI - Safe motherhood. A matter of human rights and social justice. PMID- 10670367 TI - Reproductive child health programme. An approach to reach 'Health for All by 2000 AD'. PMID- 10670368 TI - Geriatric problems and their management. The nursing perspective. AB - Geriatric problems and its Nursing care is not like any other speciality which deals with one group of diseases. Older adults suffer from various disabilities and diseases. Geriatric group will lose memory power, bearing, eyesight, etc. The Nurse who deals with geriatric patients is expected to be well versed in all the aspects. She approaches the patients through Nursing process. She plans and directs the day-to-day Nursing care according to the situation that arises. Because of the scientific knowledge and modern techniques acquired by the Nurses and health care workers they are able to give a fairly comfortable life to the elderly people. PMID- 10670369 TI - World Health Day. April 7, 1998. Safe motherhood. PMID- 10670370 TI - Building partnerships for community health. PMID- 10670371 TI - The community as main partner in health. PMID- 10670372 TI - Job stress perception among nurses. The ICU scenario. PMID- 10670373 TI - Knowledge of urban mothers about high risk conditions during pregnancy. A case study. PMID- 10670374 TI - Oxygen therapy for children. Need-based preparation and evaluation of a self instructional module for staff nurses on care of a child receiving oxygen therapy. AB - The present study was carried out with the objectives of determining the learning needs of Staff Nurses regarding care of children receiving Oxygen Therapy; finding association between learning needs and selected variables: age, total years of experience, experience in Paediatric Ward, married with or without children; determining validity of self-instructional module, on "care of child receiving Oxygen Therapy", and evaluating the effectiveness of the self instructional module or SIM. The study was conducted in two phases. A survey approach was used for Phase-I and one group pre-test post-test design was adopted for Phase-II. The total sample of the study was 30 Staff Nurses, of 6 months experience in Paediatric Ward. The findings of the study showed high learning need status in most of the areas and the Staff Nurses also expressed the desirable need for learning in detail. It was found that age, total years of experience, experience in Paediatric Ward and married with or without children were independent of their learning need. SIM was effective in terms of gain in knowledge score as well as acceptability and utility scores of Staff Nurses. PMID- 10670376 TI - The Trained Nurses' Association of India. State SNA advisors list. PMID- 10670375 TI - Awareness effect on nursing students of planned teaching programmes on AIDS. PMID- 10670377 TI - Nurse's role in pain management.2. PMID- 10670378 TI - Hygienic measures by mothers during breast feeding. PMID- 10670379 TI - Infant and mental health. PMID- 10670380 TI - Nurses as health educators. A special teachers' day feature. PMID- 10670381 TI - Nurses' role in pain management. AB - In the first part of this article (NJI, August, 1998), the author defined 'Pain' and dwelt on Pain Characteristics, Pain Components, Pain Theories and some Factors Affecting Pain. In this concluding part she describes other factors and goes on to deal with the issues regarding assessment of the pain experience and the aspect of precention of intensification of pain and the Nurses' role in providing pain relief. PMID- 10670382 TI - Holistic nursing. From knowledge of disease to knowledge of human beings. PMID- 10670383 TI - Growth pattern of children between 1 and 3 years. PMID- 10670384 TI - Nursing service administration for better patient care. A report on the TNAI Workshop. Trained Nurses' Association of India. PMID- 10670385 TI - Importance of human relations in nursing. PMID- 10670386 TI - Shaping future dietetics professionals: what's in it for you? PMID- 10670387 TI - Institute of Medicine urges Medicare coverage of medical nutrition therapy. PMID- 10670388 TI - To read or not to read original research articles: it should not be a question. PMID- 10670389 TI - Foodservice benchmarking: practices, attitudes, and beliefs of foodservice directors. AB - OBJECTIVES: To identify foodservice directors' use of performance measures and to determine their current practices of, and attitudes and beliefs about, benchmarking. DESIGN: A survey was conducted using a researcher-developed questionnaire that had been validated in a pilot-test. The questionnaire was mailed to 600 randomly selected foodservice directors; 247 (41%) responses were analyzed. SUBJECTS/SETTING: Subjects were foodservice directors in the United States from 4 categories of foodservice operations: college/university, correctional, health care, and school. STATISTICAL ANALYSES: Results were analyzed using descriptive statistics and chi 2 tests to investigate associations between variables of interest. RESULTS: The most common performance measures used by foodservice directors were food cost percentage, cost per unit or area of service, and meals per labor hour. Internal benchmarking had been used by 71% of the respondents, external benchmarking by 60%, and functional/generic by 25%. Seventy-seven percent of the respondents thought benchmarking had some or great importance in their jobs. Category of foodservice operation was associated with type of benchmarking partner and was related to certain performance measures. Sixty-one percent of respondents reported needing knowledge and skills about benchmarking. APPLICATIONS/CONCLUSIONS: Foodservice directors, regardless of category of foodservice operation, perceive benchmarking as a useful management tool to improve processes, products and services. Foodservice directors can use benchmarking to compare their financial performance with that of other organizations and learn how to improve their facility by examining best-practice processes of successful organizations. PMID- 10670390 TI - Computer-based simulations enhance clinical experience of dietetics interns. AB - OBJECTIVE: To determine the impact of computer-based simulations on the performance of dietetics interns in initial clinical rotations. DESIGN: Interns used either a simulation program (Care Planning Simulation System CPSS[) or a computer-based tutorial (Nutrition Care Planning Tutorial NCPT[) during their orientation. Performance of these interns on nutrition care skills was evaluated during their initial clinical rotations. SUBJECTS/SETTING: Participants were 108 dietetics interns from 8 different programs. Each internship had at least 8 interns, and none of the internships awarded a graduate degree. INTERVENTION: Subjects in the experimental group completed nutrition assessment and care planning activities for 3 simulated patients. Subjects in the control group completed a tutorial on assessment and care planning. MAIN OUTCOME MEASURES: Likert scale ratings of 31 behaviors were recorded by clinical preceptors. Behavior statements were grouped into 8 categories and average ratings for each category were determined. STATISTICAL ANALYSIS: Repeated-measures analysis of variance and linear regression were used to compare performance ratings between groups. RESULTS: There were no differences in overall evaluations of the simulation and tutorial groups for the 8-week period. Interns who started clinical rotations immediately after orientation (CPSS-I and NCPT-I groups) were rated lower in all categories than those who began their clinical rotations later (CPSS-D and NCPT-D groups). Maturation and acquisition of general skills likely influenced ratings of interns with delayed clinical rotations. For most categories of behavior the rate of improvement in rating scores was greatest for interns who used CPSS. APPLICATIONS: Computer-based simulations are a promising supplement to current techniques in didactic instruction and may be useful in both didactic and practice settings. Computer-based simulations can provide more varied practice experiences to didactic students and interns in preparation for more skilled entry-level positions in dietetics. PMID- 10670391 TI - Genetic taste markers and preferences for vegetables and fruit of female breast care patients. AB - OBJECTIVE: To explore links between genetic responsiveness to the bitter taste of 6-n-propylthiouracil (PROP) and self-reported preferences for vegetables and fruit of female breast care patients. METHODS: PROP tasting was defined by detection thresholds and by perceived bitterness and hedonic ratings for PROP solutions. Nontasters, medium tasters, and supertasters were identified by their PROP thresholds and by the ratio of perceived bitterness of PROP to the perceived saltiness of sodium chloride solutions. Subjects rated preferences for vegetables and fruit using 9-point category scales. SUBJECTS/SETTING: A clinical sample of 170 patients with newly diagnosed breast cancer and 156 cancer-free control subjects were recruited from the University of Michigan Breast Care Center. STATISTICAL ANALYSES: Principal components factor analysis, one-way analyses of variance, and Pearson correlations and chi 2 tests were used to analyze taste and food preference data. RESULTS: Genetic responsiveness to PROP was associated with lower acceptance of cruciferous and selected green and raw vegetables (P < .05). Women who reported disliking such foods were medium tasters or supertasters of PROP. Preference ratings for fruit were unrelated to PROP taster status. APPLICATIONS/CONCLUSIONS: Women who are PROP tasters may be less likely to comply with dietary strategies for cancer prevention that emphasize consumption of cruciferous vegetables and bitter salad greens. Alternatively, PROP-sensitive women may seek to reduce bitter taste by adding fat, sugar, or salt. PMID- 10670392 TI - Early adopters of olestra-containing foods: who are they? AB - OBJECTIVE: To identify the characteristics of people consuming olestra-containing foods when first introduced at a test-marketing site. DESIGN: Data are from the Olestra Postmarketing Surveillance Study (OPMSS). After the introduction of olestra into a large test-marketing site, study participants received 3 follow-up telephone calls, at 3-month intervals, in which they were questioned about their diets during the previous month. SUBJECTS/SETTING: 1,007 adults in Indianapolis, Ind, who participated in a baseline clinic visit (before introduction of olestra into the food market) and completed at least 2 of 3 follow-up telephone calls (after the introduction of olestra into the market). STATISTICAL ANALYSES PERFORMED: Logistic regression was used to examine associations between olestra consumption and sociodemographic characteristics, health conditions, attitudes toward health and diet, and health-related behaviors. RESULTS: Olestra consumption on at least 1 of the follow-up telephone calls was reported by 41.5% of the study sample, and consumption on 2 or more telephone calls was reported by 20.0% of the sample. Factors associated with early adoption of olestra-containing foods included white ethnicity, higher education, overweight, absence of diabetes, attitudes indicative of diet and health concerns (e.g.; perceptions that there is a strong relationship between diet and disease), and a lower fat intake. APPLICATIONS/CONCLUSIONS: In spite of the controversy surrounding the introduction of olestra into the food market persons with attitudes indicative of diet and health concerns were likely to be early adopters of olestra-containing foods. Dietitians and other health care providers should inquire about intake levels of foods with fat substitutes and ensure that these foods are not being consumed in excessive amounts or being consumed instead of nutrient-dense foods that are naturally low in fat. PMID- 10670393 TI - Assessing food selection in a health promotion program: validation of a brief instrument for American Indian children in the southwest United States. AB - OBJECTIVE: Brief dietary assessment instruments are needed to evaluate behavior changes of participants in dietary intervention programs. The purpose of this project was to design and validate an instrument for children participating in Pathways to Health, a culturally appropriate, cancer prevention curriculum. DESIGN: Validation of a brief food selection instrument, Yesterday's Food Choices (YFC), which contained 33 questions about foods eaten the previous day with response choices of yes, no, or not sure. Reference data for validation were 24 hour dietary recalls administered individually to 120 students selected randomly. SUBJECTS: The YFC and 24-hour dietary recalls were administered to American Indian children in fifth- and seventh-grade classes in the Southwest United States. STATISTICAL ANALYSES PERFORMED: Dietary recalls were coded for food items in the YFC and results were compared for each item using percentage agreement and the kappa statistic. RESULTS: Percentage agreement for all items was greater than 60%; for most items it was greater than 70%, and for several items it was greater than 80%. The amount of agreement beyond that explained by chance (kappa statistic) was generally small. Three items showed substantial agreement beyond chance (kappa > or = 0.6); 2 items showed moderate agreement (kappa = 0.40 to 0.59) most items showed fair agreement (kappa = 0.20 to 0.39). The food items showing substantial agreement were hot or cold cereal, low-fat milk, and mutton or chile stew. Fried or scrambled eggs and deep-fried foods showed moderate agreement beyond chances. CONCLUSIONS: Previous development and validation of brief food selection instruments for children participating in health promotion programs has had limited success. In this study, instrument-related factors that apparently contributed to poor agreement between data from the YFC and 24-hour dietary recall were inclusion of categories of foods vs specific foods; food knowledge, preparation, and vocabulary, item length, and overreporting of attractive foods. Collecting and scoring the 24-hour recall data may also have contributed to poor agreement. Further development of brief instruments for evaluating changes in children's behavior in dietary programs is necessary. Factors related to the YFC that need further development may be issues that are also important in the development of effective, brief dietary assessments for children as individual clients or patients. PMID- 10670394 TI - Randomized multicenter trial documenting the efficacy and safety of a lactose free and a lactose-containing formula for term infants. AB - OBJECTIVE: To evaluate the efficacy and safety of a new lactose-free infant formula. DESIGN: Randomized, prospective, double-blind, controlled, outpatient, multicenter, parallel 12-week trial. SETTING: Ambulatory-care facilities of the participating centers. SUBJECTS: 137 healthy term infants (approximately 7 days old at the time of study enrollment). INTERVENTION: Healthy term infants, whose mothers had decided not to breast-feed, were randomly assigned 1 of the 2 study formulas. MAIN OUTCOME MEASURES: Weight, length, and occipitofrontal circumference measurements were obtained at baseline and when the infant was 2, 4, 8, and 12 weeks old. Formula acceptance and tolerance were also assessed at weeks 2, 4, 8, and 12. Serum albumin concentration, creatirune level, and blood urea nitrogen were determined at baseline and week 12. Adverse events were assessed throughout the study. STATISTICAL ANALYSES PERFORMED: Each baseline anthropometric and laboratory variable was analyzed for comparability between groups using the Student t test and was also analyzed using a repeated-measures analysis of variance method. Covariance analysis was applied to the final laboratory data using the respective baseline data as covariates. Decisions about equality of mean responses to formula effects were based on the .05 level of significance in all cases. RESULTS: One hundred four infants completed the study. No significant differences between the 2 formula groups were noted for any of the growth and blood parameters. APPLICATIONS: This new formula is an effective and safe lactose-free nutrition alternative for infants who require such a diet. PMID- 10670395 TI - Dietitian intervention improves lipid values and saves medication costs in men with combined hyperlipidemia and a history of niacin noncompliance. AB - We asked if medical nutrition therapy (MNT) administered by registered dietitians could lead to beneficial clinical and financial outcomes in men with combined hyperlipidemia (hypercholesterolemia and hypertriglyceridemia). A retrospective chart review was conducted on 73 men with combined hyperlipidemia who were being considered for statin therapy because of a previous history of noncompliance with niacin therapy. Subjects participated in an 8-week dietitian intervention program as a qualifying requirement, before statin therapy. Patient records were reviewed to determine the beginning and ending serum lipid concentrations and the number and length of dietitian sessions. Complete information was available on 43 subjects, aged 60.7 +/- 10.1 years (mean +/- standard deviation). Total dietitian intervention time was 169 +/- 19 minutes in 2.7 +/- 0.6 sessions (range = 2 to 4 sessions) during 6.5 +/- 2.2 weeks of MNT (range = 4 to 8 weeks). MNT lowered levels of total cholesterol 11% (P < .001), low-density lipoprotein cholesterol 9% (P < .001), and triglycerides 22% (P < .0001) and body mass index 2% (P < .0001); MNT raised high-density lipoprotein cholesterol levels 4%. After dietitian intervention, only 15 of 30 eligible patients required antihyperlipidemic medications, which led to an annual cost savings of $27,449.10 or $638.35 per patient. A cost saving of $3.03 in statin therapy was realized for each dollar spent on MNT. We conclude that an average of 3 individualized dietitian visits of 1 hour each over an 8-week period has a beneficial effect in treating patients with combined hyperlipidemia and recommend consideration of MNT as a cost-effective intervention. PMID- 10670396 TI - Diabetes training for dietitians: needs assessment, program description, and effects on knowledge and problem solving. AB - Recent changes in management and medical nutrition therapy for diabetes mellitus have produced a need to retrain many practicing dietitians. To meet this need, a multidisciplinary group experienced in medical nutrition therapy and educational methods used a formal needs-assessment process to design a new training program. Sugar is Not a Poison (SNAP): The Dietitian's New Role in Diabetes Management is a 2 1/2-day program that uses written text, didactic presentation, and exercises that simulate patient encounters to accomplish 12 learning objectives. Program evaluations show high levels of participant satisfaction. Mean (+/- standard deviation) scores on pre- and postests of knowledge and problem solving were 69 +/- 13% and 86 +/- 9%, respectively (P < 0.01). The SNAP program needs assessment, training methods, and knowledge problem-solving test are relevant to all types of education programs in clinical dietetics. PMID- 10670397 TI - Caffeine intake in young children differs by family socioeconomic status. PMID- 10670398 TI - Serving portion size influences 5-year-old but not 3-year-old children's food intakes. PMID- 10670399 TI - Overconcern about thinness in 10- to 14-year-old schoolgirls in Taiwan. PMID- 10670400 TI - Comparison of body weight and body fat classifications of competitive school-age club swimmers. PMID- 10670401 TI - Validity and reliability of qualitative dietary fat index questionnaires: a review. PMID- 10670402 TI - Ground ostrich: a comparison with ground beef. PMID- 10670403 TI - Position of the American Dietetic Association: food irradiation. AB - Food irradiation has been identified a sa safe technology to reduce the risk of foodborne illness as part of high-quality food production, processing, handling, and preparation. Food irradiation's history of scientific research , evaluation, and testing spans more than 40 countries around the world and it has been endorsed or support by numerous national and international food and organizations and professional groups. Food irradiation utilizes a source of ionizing energy that passes through food to destroy harmful bacteria and other organism. Often referred to as "cold pasteurization," food irradiation offers negligible loss of nutrients or sensory qualities in food as it does not substantially raise the temperature of the food during processing. Food irradiation does not replace proper food production, processing, handling, or preparation, nor can it enhance the quality of or prevent contact with foodborne bacteria after irradiation. In the United States, manufacturers are required to identify irradiated food sold to consumers with an international symbol (Radura) and and terminology describing the process on product labels. In addiction, food irradiation facilities are thoroughly regulated and monitored for worker and environmental safety. Members of The American Dietetic Association (ADA) and other food, nutrition, and health professionals have a responsibility to educate consumers, food processors, manufacturers and retailers about the safety and application of the technology. When consumers are educated about food irradiation, many prefer irradiated products because of their increased safety. It is the position of ADA that food irradiation enhances the safety and quality of the food supply and helps protect consumers from foodborne illness. The ADA encourages the government, food manufactures, food commodity groups, and qualified food and nutrition professionals to work together to educate consumers about this additional food safety tool and make this choice available in the marketplace. PMID- 10670404 TI - Case problem: presenting conventional and complementary approaches for relieving nausea in a breast cancer patient undergoing chemotherapy. PMID- 10670405 TI - Severe hypophosphatemia. Pathophysiologic implications, clinical presentations, and treatment. AB - We conducted this review to heighten the awareness and describe pathologic manifestations of hypophosphatemia. We present 3 cases of varied manifestations of hypophosphatemia where recognition was delayed. In certain settings, severe hypophosphatemia has significant morbidity and potential mortality. Appreciation of the pathophysiologic basis for organ dysfunction in severe hypophosphatemia should result in early recognition and treatment. We reviewed the English language literature for reported cases and research studies dealing with pathophysiologic mechanisms subserving clinical manifestations. We observed that depletion of adenosine triphosphate (ATP) would explain most of the derangement noted in cellular functions. Phosphate plays a key role in the delivery of oxygen to the tissue. Lack of phosphate, therefore, leads to tissue hypoxia and hence disruption of cellular function. Severe hypophosphatemia becomes clinically significant when there is underlying phosphate depletion. Otherwise, short-term acute hypophosphatemia is not usually associated with any specific disorder. Chronic hypophosphatemia, on the other hand, results in hematologic, neuromuscular, and cardiovascular dysfunction, and unless corrected, the consequences can be grave. Most of the time hypophosphatemia results from renal loss of phosphate, diagnosed by a fractional secretion of phosphate > 5%. It is hard to provide precise estimates of how many patients are seen with hypophosphatemia annually at academic medical centers. This is complicated by use of chemistry panels that do not measure inorganic phosphate unless specifically ordered. This often leads to delay in correct diagnosis, and, therefore, additional delay in providing appropriate management. A high index of suspicion alone avoids the unnecessary withholding of treatment that can be life saving. PMID- 10670406 TI - Assessing the effects of thyroid suppression on benign solitary thyroid nodules. A model for using quantitative research synthesis. AB - Systematic review of the available information with a modified, largely quantitative method of research synthesis disclosed that an initial trial of thyroid hormone suppression therapy leads to clinically significant (> or = 50%) reduction of nodule size or arrest of nodule growth in a subset of patients with benign solitary thyroid nodules. In fact, in addition to objective improvements due to decreasing nodule size, L-T4 suppression therapy may benefit patients by reducing perinodular thyroid volume. Consequently, both pressure symptoms and cosmetic complaints may improve (9, 68). Additional studies for the assessment of the risks versus benefits of supraphysiologic doses of L-T4, the optimal level of thyroid suppression and the dose needed to achieve this magnitude of reduction, the optimal length of the initial trial, and the conditions for the continuation of L-T4 thyroid suppression therapy, as well as the identification of markers for patients most likely to respond to this therapy, are warranted. Finally, quantitative assessment of available evidence as described here may be applicable to the review of other controversial issues as well. PMID- 10670407 TI - The adult patient with Ebstein anomaly. Outcome in 72 unoperated patients. AB - Knowledge of the long-term outcome in unoperated adult patients with Ebstein anomaly is limited, and the therapeutic approach is still controversial. We studied unoperated adult patients with Ebstein anomaly to define the patterns of presentation, anatomic characteristics, outcome, and predictive factors for survival. Seventy-two unoperated survivors of Ebstein anomaly aged over 25 years attended from 1972 to 1997 were reviewed and followed-up from 1.6 to 22.0 years. Patients were classified in 3 groups of severity according to the echocardiographic appearance of the septal leaflet attachment of tricuspid valve. The mean age at diagnosis was 23.9 +/- 10.4 years, and the most common clinical presentation was an arrhythmic event (51.4%). There were 30 (42%) deaths, including 6 from arrhythmia, 12 related to heart failure, 7 sudden, 2 unrelated, and 3 unascertained. According to Cox regression analysis, predictors of cardiac related death included age at diagnosis (hazard ratio 0.89 for each year of age, 95% confidence intervals CI[ 0.84-0.94), male sex (3.93, 95% CI, 1.50-10.29), degree of echocardiographic severity (3.34, 95% CI, 1.78-6.24), and cardiothoracic ratio > or = 0.65 (3.57, 95% CI, 1.15-11.03). During follow-up, morbidity was mainly related to arrhythmia and refractory late hemodynamic deterioration. The magnitude of tricuspid regurgitation, cyanosis, and the New York Heart Association (NYHA) functional class at time zero were significant risk factors according to the univariate analysis, but not after multivariable confrontation. The results of this study suggest that pattern of presentation, clinical course, and prognosis of unoperated adult patients with Ebstein anomaly are influenced by several factors. Although the initial symptoms are usually mild and commonly related to supraventricular arrhythmias, these are not associated with the long-term outcome. The severity of the morbid anatomy was the main determinant of survival only in extreme cases, but not in those with mild or moderate deformations, which are more common in adults. Other independent risk factors such as cardiothoracic ratio, sex, age at diagnosis, and the echocardiographic evaluation may help to determine the therapeutic approach. Adult patients with Ebstein anomaly should not be considered as a simple low-risk group. PMID- 10670408 TI - Pyoderma gangrenosum. A comparison of typical and atypical forms with an emphasis on time to remission. Case review of 86 patients from 2 institutions. AB - Pyoderma gangrenosum (PG) is an idiopathic, inflammatory, ulcerative disease of undetermined cause. The diagnosis is based on clinical and pathologic features and requires exclusion of conditions that produce ulcerations. An atypical bullous variant (atypical pyoderma gangrenosum, APG) exists with clinical features similar to those of Sweet syndrome. Because PG is a rare disease, few large case series have been reported. Pyoderma gangrenosum was first recognized as a unique disease entity in the first half of the 20th century. Cumulative knowledge of PG is based on a handful of case series and multiple individual case reports. To augment that knowledge, we present our experience with a large number of patients over a significant time. We performed a retrospective analysis of the medical records of 86 patients with PG who were evaluated and treated over 12 years at 2 university-based dermatology departments. The mean (+/- standard deviation) age of onset of PG and APG, respectively, was 44.6 +/- 19.7 years and 52.2 +/- 15.3 years. Lower extremity involvement was most common in PG, whereas upper extremity involvement was most common in APG. Associated relevant systemic diseases were seen in 50% of patients. Inflammatory bowel disease was the most common association in patients with PG, whereas hematologic disease or malignancy was most common in those with APG. Although a few patients were managed with local measures or nonimmunosuppressive treatment, the majority required oral corticosteroid therapy, often with systemic immunosuppressive treatment. PG patients required a mean 11.5 +/- 11.1 months of treatment to achieve remission compared with 9.0 +/- 13.7 months for patients with APG. Five patients (5.8%) had disease that was extremely refractory to multiple intensive therapies. The prognosis and disease associations for PG and APG appear to be different. Compared with PG, APG is more often associated with hematologic disease or malignancy, and remits more quickly. PMID- 10670409 TI - Extrahepatic manifestations associated with hepatitis C virus infection. A prospective multicenter study of 321 patients. The GERMIVIC. Groupe d'Etude et de Recherche en Medecine Interne et Maladies Infectieuses sur le Virus de l'Hepatite C. AB - From January 1996 to January 1997, 321 patients with an average age of 46 +/- 16 years and chronically infected with hepatitis C virus (HCV) were prospectively enrolled in a study designed to determine the prevalence of extrahepatic manifestations associated with HCV infection in a large cohort of HCV patients, to identify associations between clinical and biologic manifestations, and to compare the results obtained in human immunodeficiency virus (HIV)-positive versus HIV-negative subsets. In a cross-sectional study, clinical extrahepatic manifestations, viral coinfections with HIV and/or hepatitis B virus, connective tissue diseases, and a wide panel of autoantibodies were assessed. Thirty-eight percent (122/321) of patients presented at least 1 clinical extrahepatic manifestation including arthralgia (60/321, 19%), skin manifestations (55/321, 17%), xerostomia (40/321, 12%), xerophthalmia (32/321, 10%), and sensory neuropathy (28/321, 9%). Main biologic abnormalities were mixed cryoglobulins (110/196, 56%), thrombocytopenia (50/291, 17%), and the presence of the following autoantibodies: antinuclear (123/302, 41%), rheumatoid factor (107/280, 38%), anticardiolipin (79/298, 27%), antithyroglobulin (36/287, 13%) and antismooth muscle cell (27/288, 9%). At least 1 autoantibody was present in 210/302 (70%) of sera. By multivariate logistic regression analysis, 4 parameters were significantly associated with cryoglobulin positivity: systemic vasculitis (p = 0.01, odds ratio OR[ = 17.3), HIV positivity (p = 0.0006, OR = 10.2), rheumatoid factor positivity (p = 0.01, OR = 2.8), and sicca syndrome (p = 0.03, OR = 0.27). A definite connective tissue disease was noted in 44 patients (14%), mainly symptomatic mixed cryoglobulinemia and systemic vasculitis, HIV coinfection (23%) was associated with 3 parameters: anticardiolipin (p = 0.003, OR = 4.18), thrombocytopenia (p = 0.01, OR = 3.56), and arthralgia or myalgia (p = 0.017, OR = 0.23). HIV-positive patients presented more severe histologic lesions (p = 0.0004). Extrahepatic clinical manifestations in HCV patients involve primarily the skin and joints. The most frequent immunologic abnormalities include mixed cryoglobulins, rheumatoid factor, antinuclear, anticardiolipin, and antithyroglobulin antibodies. Cryoglobulin positivity is associated with systemic vasculitis and rheumatoid factor and HIV positivity. HIV coinfection is associated with arthralgia or myalgia, anticardiolipin antibodies, and thrombocytopenia. PMID- 10670410 TI - Can neurologic manifestations of Hughes (antiphospholipid) syndrome be distinguished from multiple sclerosis? Analysis of 27 patients and review of the literature. AB - Hughes (antiphospholipid) syndrome (APS) can mimic multiple sclerosis (MS). We analyzed the clinical, laboratory, and imaging findings of MS-like expression in a cohort of patients with APS in an attempt to identify parameters that might differentiate the 2 entities. We studied 27 patients who were referred to our unit with the diagnosis of probable or definite MS made by a neurologist. All patients were referred to our lupus clinic because of symptoms suggesting an underlying connective tissue disease, uncommon findings for MS on magnetic resonance imaging (MRI), atypical evolution of MS, or antiphospholipid antibody (aPL) positivity. aPL, antinuclear antibody (ANA), anti-dsDNA, and anti extractable nuclear antigen (ENA) antibodies were measured by standard methods. MRI was performed in every patient and compared with MRI of 25 definite MS patients who did not have aPL. An index severity score was calculated based on the size and number of increased signal intensity areas in MRI. In the past medical history, 8 patients with primary APS and 6 with APS secondary to systemic lupus erythematosus (SLE) had had symptoms related to these conditions. Neurologic symptoms and physical examination of the patients were not different from those common in MS patients. Laboratory findings were not a useful tool to distinguish APS from MS. When MRI from APS patients was compared globally with MRI from MS patients, MS patients had significantly increased severity score in white matter (p < 0.001), cerebellum (p = 0.035), pons (p < 0.015), and when all areas were taken together (p < 0.001). Patients with APS had significantly increased scores in the putamen (p < 0.01). No differences were noticed in the degree of atrophy. When taken individually, MRI from APS patients could not be distinguished from MRI from MS patients. Most of the patients with primary APS showed a good response to oral anticoagulant treatment. In patients with secondary APS, the outcome was poorer. Hughes syndrome (APS) and MS can be difficult to distinguish. A careful medical history, a previous history of thrombosis and/or fetal loss, an abnormal localization of the lesions in MRI, and the response to anticoagulant therapy might be helpful in the differential diagnosis. We believe that testing for aPL should become routine in all patients with MS. PMID- 10670411 TI - Delta-opioid receptor-mediated increase in cortical extracellular levels of cholecystokinin-like material by subchronic morphine in rats. AB - Numerous pharmacological data indirectly support the idea that interactions between cholecystokinin (CCK) and opioids participate in the development of tolerance to morphine. Biochemical investigations were performed with the aim of directly assessing the status of such interactions in morphine treated rats. Tolerance to the alkaloid after s.c. implantation of morphine pellets for three days was not associated with any change in the levels of both CCK like-material (CCKLM) and proCCK mRNA in the frontal cortex. However, microdialysis in the freely moving rat showed that this morphine treatment produced a significant increase (+40%) of the cortical spontaneous CCKLM outflow, which could be completely prevented by intracortical infusion of naloxone (10 microM). The opioid receptors responsible for morphine-induced cortical CCKLM overflow appeared to be of the delta type because intracortical infusion of selective delta-opioid receptor antagonists such as naltriben (10 microM) and 7 benzylidenenaltrexone (10 microM) also prevented the effect of morphine, whereas CTOP (10 microM), a selective mu-opioid receptor antagonist, and nor binaltorphimine (10 microM), a selective K-opioid receptor antagonist, were inactive. These data indicate that morphine tolerance is associated with delta opioid receptor mediated activation of cortical CCKergic systems in rats. PMID- 10670412 TI - Pharmacological comparison of P2X receptors on rat coeliac, mouse coeliac and mouse pelvic ganglion neurons. AB - Characteristics of P2X receptors on neurons of the rat coeliac, mouse coeliac and mouse pelvic ganglia have been studied using the whole cell voltage-clamp technique. Fast application of ATP (100 microM) on to isolated neurons voltage clamped at -70 mV induced a slowly desensitising inward current in 96% of the cells tested. Concentration-response curves for ATP yielded EC50 values of 86 microM, 64 microM and 123 microM, for rat coeliac, mouse coeliac and mouse pelvic ganglion neurons, respectively, while alpha,beta-methylene ATP was inactive. The response to ATP was antagonised by suramin, Cibacron blue and pyridoxalphosphate 6-azophenyl-2',4'-disulphonic acid (PPADS). The potency of ATP was increased by extracellular acidification and by co-application of micromolar concentrations of Zn2+, while raising pH decreased it. On rat coeliac ganglion neurons, the EC50 values for ATP were 35 microM and 253 microM at pH 6.8 and 8.0, respectively. On mouse coeliac and pelvic ganglion neurons, altering the pH produced comparable changes. In conclusion, our results indicate that, in contrast to the guinea-pig coeliac ganglion, the characteristics of the P2X receptors present on rat coeliac, mouse coeliac and mouse pelvic ganglia are all identical to those present on rat pelvic ganglion, i.e. they are homomeric P2X2 receptors, or heteromultimers with P2X2 being the dominant subunit. PMID- 10670413 TI - The prostaglandin E series modulates high-voltage-activated calcium channels probably through the EP3 receptor in rat paratracheal ganglia. AB - The modulation of high-voltage-activated (HVA) Ca2+ channels by the prostaglandin E series (PGE1 and PGE2) was studied in the paratracheal ganglion cells. Prostaglandin E1, E2, STA2 (a stable analogue of thromboxane A2), 17-phenyl trinor-PGE2 (an EP1-selective agonist) and sulprostone (an EP3-selective agonist) inhibited the HVA Ca2+ current (HVA ICa) dose-dependently, and the rank order of potency to inhibit HVA Ca2+ channels was sulprostone>PGE2, PGE1>STA2>>17-phenyl trinor-PGE2. SC-51089 (10(-5) M), a selective EP1-receptor antagonist, showed no effect on the PGE1- or PGE2-induced inhibition of the HVA ICa, thereby indicating that PGE1- and PGE2-induced inhibition of the HVA Ca2+ channels is possibly mediated by the EP3 receptor. The PGE1-sensitive component of the current was markedly reduced in the presence of omega-conotoxin-GVIA (3x10(-6) M), but not with nifedipine (3x10(-6) M). PGE1 and PGE2 also inhibited the remaining ICa in a saturating concentration of nifedipine, omega-conotoxin-GVIA and omega-conotoxin MVIIC, suggesting that R-type Ca2+ channels are involved. The inhibitory effect of PGE1 or sulprostone was prevented by pretreatment with pertussis toxin [islet activating protein (IAP)] or phorbol-12-myristate-13-acetate (PMA), and the protein kinase C (PKC) inhibitor chelerythrine blocked the action of PMA. It was concluded that PGE1 selectively reduces both N- and R-type Ca2+ currents by activating a G-protein probably through the EP3 receptor in paratracheal ganglion cells. PMID- 10670414 TI - Dopamine activates inward rectifier K+ channel in acutely dissociated rat substantia nigra neurones. AB - The effect of dopamine (DA) was investigated on acutely dissociated rat substantia nigra pars compacta (SNc) neurones by using patch clamp recording. The SNc neurones could be classified into two groups. About 75% of large neurones (>30 microm in diameter) were tyrosine hydroxylase (TH) positive while almost all small neurones (<20 microm) were TH negative. In the large neurones, DA hyperpolarized the membrane, resulting in a reduction of the frequency of spontaneous action potentials in current-clamp mode and induced an inward rectifier K+ current in voltage-clamp mode. Quinpirole, a D2 receptor agonist, mimicked the DA action. S(-)-sulpiride, a D2 receptor antagonist, inhibited the DA-induced current (I(DA)) more effectively than SKF83566, a D1 receptor antagonist. Intracellular application of either guanosine 5'-O-(2 thiodiphosphate) (GDP-betaS) or pertussis toxin (IAP) suppressed I(DA). Guanosine 5'-O-(3-thiotriphosphate) (GTP-gammaS) sustained the DA response. Modulators for cAMP such as forskolin and isobutylmethylxathine, H-89, a protein kinase A inhibitor, and chelerythrine, a protein kinase C inhibitor, had no effect on I(DA). The frequency of DA-induced single channel currents in the inside-out patch configuration, for which the unitary conductance was 56.6pS, was greatly reduced by the replacement of GTP with GDP perfused at the cytosolic side. These results suggest that DA acts on a D2-like receptor and activates directly an IAP sensitive G protein coupled with inward rectifier K+ channels, resulting in a decrease in the spontaneous firing activities of rat SNc dopaminergic neurones. PMID- 10670415 TI - Sensitivity of native and cloned hippocampal delayed-rectifier potassium channels to verapamil. AB - The effects of the phenylalkylamine verapamil on native and cloned hippocampal voltage-operated potassium channels were investigated. Native channels were studied in acutely isolated CA1 neurons from the guinea pig with the whole-cell patch-clamp technique. Cloned channels were expressed in oocytes of Xenopus laevis and studied with the two-electrode voltage-clamp technique. Native potassium channels: Verapamil suppressed the potassium currents in micro- and submicromolar concentrations. The current suppression increased during the voltage step. The IC50 value of verapamil was 3 micromol/l and the Hill coefficient was 0.5 indicating a mixed population of potassium channels with distinct verapamil sensitivity. Cloned potassium channels: The hippocampal potassium channels Kv1.1, Kv1.2, Kv1.3, Kv2.1, Kv3.1 and Kv3.2 were affected by verapamil in micromolar concentrations. The effect increased with depolarization time, was voltage-dependent, reached 90% of the maximum within around 40 s after start of verapamil application, recovered slowly after wash-out and did not reach control values even after wash-out times of six minutes. The IC50 values differed markedly and were 35 micromol/l for the Kv1.1 channel, 98 micromol/l for the Kv1.2 channel, 12 micromol/l for the Kv1.3 channel, 226 micromol/l for the Kv2.1 channel, 6 micromol/l for the Kv3.1 channel and 11 micromol/l for the Kv3.2 channel. PMID- 10670416 TI - Neuroprotective effects of an AMPA receptor antagonist YM872 in a rat transient middle cerebral artery occlusion model. AB - The neuroprotective effects of YM872 ([2,3-dioxo-7-(1H-imidazol-1-yl)6-nitro 1,2,3,4-tetrahydro-1-quinoxal inyl]acetic acid monohydrate), a novel alpha-amino 3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor antagonist with high water solubility, were examined in rats with transient middle cerebral artery (MCA) occlusion. The right MCA of male SD rats was occluded for 3 h using the intraluminal suture occlusion method. YM872 significantly reduced the infarct volume 24 hours after occlusion, at dosages of 20 and 40 mg/kg/h (iv infusion) when given for 4 h immediately after occlusion. Furthermore, delayed administration of YM872 (20 mg/kg/h iv infusion for 4 h, starting 2 or 3 h after the occlusion) also reduced the infarct volume and the neurological deficits measured at 24 h. Additionally, the therapeutic efficacy of YM872 persisted for at least seven days after MCA occlusion in animals treated with YM872 for 4 h starting 2 h after MCA occlusion. These data demonstrate that AMPA receptors contribute to the development of neuronal damage after reperfusion as well as during ischemia in the focal ischemia models and that the acute effect of the blockade of AMPA receptors persists over a long time period. YM872 shows promise as an effective treatment for patients suffering from acute stroke. PMID- 10670417 TI - Dynamics of NMDAR-mediated neurotoxicity during chronic ethanol exposure and withdrawal. AB - We have utilized a hippocampal brain slice explant system to assess cellular and synaptic mechanisms underlying the expression of alcohol withdrawal hyperexcitability. Previously, we observed a role for NMDA receptors in the expression of electrographic seizures (EGS) observed immediately upon withdrawal from chronic ethanol exposure in this system. One possible cellular mechanism responsible for these prior results involves NMDAR-mediated neurotoxicity, which was assessed in the present study. Explants were exposed to 35 or 75 mM ethanol for 6 or 12 days and incubated with propidium iodide (PI) to label non-viable cells and then imaged digitally. PI labeling was significantly reduced (36% of control levels) following chronic ethanol exposure (75 mM). When tested following ethanol withdrawal, PI labeling remained significantly reduced in the 75 mM exposed group. We next assessed the effect of an NMDA challenge 24 h following withdrawal. The 35 mM and 75 mM ethanol exposed groups displayed significant 6 fold and 13-fold NMDAR-mediated increases in PI labeling respectively; control explants displayed a 3-fold increase. These data suggest that chronic ethanol exposure prior to withdrawal has a minor neuroprotective effect that slightly diminishes within 24 h of ethanol withdrawal. Furthermore, the data indicate that direct NMDAR activation is required for induction of ethanol withdrawal neurotoxicity. PMID- 10670418 TI - Chronic modulation of the GABA(A) receptor complex regulates Y1 receptor gene expression in the medial amygdala of transgenic mice. AB - NPY exerts anxiolytic effects, which are mediated by activation of Y1 receptors in the amygdala. It has been shown that diazepam counteracts the anxiogenic effect of Y1 receptor antagonists, suggesting that NPYergic and GABAergic systems are coupled in the regulation of anxiety. We used a transgenic mouse model, expressing a mouse Y1 receptor-beta-galactosidase fusion gene (Y1R/LacZ), to study the effect of positive or negative modulators of GABA(A) receptors on Y1 receptor gene expression. Mice were treated for 14 days with diazepam (4 or 20 mg/kg), the anxiolytic beta-carboline-derivative abecarnil (0.3 or 6 mg/kg) and the anxiogenic beta-carboline FG7142 (20 mg/kg). Transgene expression was determined by quantitative analysis of beta-galactosidase histochemical staining in the medial amygdala and in the medial habenula as a control region. Chronic treatment with 20 mg/kg diazepam or 6 mg/kg abecarnil significantly increased, whereas FG 7142 decreased, transgene expression in the medial amygdala. A transient decrease in transgene expression was observed in the medial amygdala six hours after the acute treatment with 20 mg/kg FG 7142 but not with diazepam or abecarnil. No significant changes were observed in the medial habenula. These data suggest that modulation of GABA(A) receptor function may regulate Y1 receptor gene expression in medial amygdala. PMID- 10670419 TI - Loreclezole inhibition of recombinant alpha1beta1gamma2L GABA(A) receptor single channel currents. AB - Loreclezole had two different effects on GABA(A) receptor (GABAR) currents. When applied to GABARs that contained a beta2 or beta3 subunit subtype, but not a beta1 subtype, loreclezole potentiated the peak current evoked by sub-maximal concentrations of GABA. Loreclezole also increased the rate and degree of apparent desensitization of GABAR whole-cell currents, an effect that was independent of the beta subunit subtype, suggesting that potentiation and inhibition of GABAR current by loreclezole occurred through separate sites. We used patch-clamp recording from outside-out and inside-out patches from L929 fibroblasts transiently transfected with rat GABAR subunits to examine the properties of inhibition of alpha1beta1gamma2L single channel currents by loreclezole. Loreclezole decreased the mean open time of the channel by decreasing the average durations of the open states. Loreclezole also increased the occurrence of a closed component with an average duration near 20 ms. Inhibition by loreclezole was not voltage-dependent. Loreclezole was equally effective when applied to the intracellular side of the receptor, suggesting that its binding site was readily accessible from both sides of the membrane. Pre application of loreclezole effectively inhibited the GABAR current in macropatches, indicating that binding did not require an open channel. These findings were consistent with a mechanism of allosteric modulation at a site formed by the membrane spanning regions of the receptor. PMID- 10670420 TI - Impaired inhibition of epileptiform activity by baclofen, but not by adenosine in the weaver hippocampus. AB - The weaver defect results in a loss of baclofen- and adenosine-gated K+ conductance in the hippocampus of adult homozygous (wv/wv) mice. In addition, suppression of hippocampal epileptiform activity by baclofen is impaired (Jarolimek, W., Baurle, J., Misgeld, U., 1998. Pore mutation in a G protein-gated inwardly rectifying K+ channel subunit causes loss of K+ dependent inhibition in weaver hippocampus. Journal of Neuroscience 18, 4001-4007). We used wv/wv and wild-type (+/+) mice to determine whether K+ conductance increases are essential for the suppression of epileptiform activity by R-baclofen and adenosine in disinhibited hippocampal slices. In wv/wv mice R-baclofen was less potent by two orders of magnitude in reducing the frequency of spontaneous synchronous burst discharges than in +/+ mice. Endogenous adenosine and adenosine A1 receptor agonists differed only slightly in their efficacy to inhibit spontaneous synchronous burst discharges in wv/wv and +/+ mice. The findings on adenosine A1 receptors suggest that the varied efficacy of R-baclofen in wv/wv and +/+ mice may not be explained solely on the basis of a loss of ligand-gated K+ conductance. Therefore, we investigated the affinity of GABA(B) receptors for the antagonist CGP55845A in wv/wv and +/+ hippocampi. Schild plot analysis revealed a K(D) for the GABA(B) antagonist CGP55845A 10 fold higher in wv/wv than in +/+ mice. The data suggest that an alteration of GABA(B) receptors could contribute to the reduced efficacy of R-baclofen to suppress hippocampal epileptiform activity in weaver mice, while the suppression by adenosine remains largely unaffected. PMID- 10670421 TI - Effects of valproate derivatives I. Antiepileptic efficacy of amides, structural analogs and esters. AB - Derivatives of the antiepileptic drug valproate (VPA, 2-propylpentanoic acid) have been synthesized and tested in order to improve the intracellular availability of VPA. The buccal ganglia of Helix pomatia were used as a test nervous system and antiepileptic efficacies were reconfirmed using rat cortex in vivo. Epileptiform activities consisted of typical paroxysmal depolarization shifts (PDS) which appeared in the identified neuron B3 with application of pentylenetetrazol. Epileptiform activities were found to be accelerated, unaffected or blocked. (i) The Amide-derivatives 2-propylpentanamide and N,N dipropyl-2-propylpentanamide, and short chain ester derivatives 1-O-(2 propylpentanoyl)-2,3-propandiol, 2,2-di(hydroxymethyl)-1-O-(2-propylpentanoyl) 1,3-propanediol and 2,2-di(hydroxymethyl)-1,3-di-O-(2-propylpentanoyl)-1,3 propanediol accelerated epileptiform activities. Membrane potential often shifted to a permanent depolarization which corresponded to the PDS-inactivation level. (ii) The structural analogs 1-cycloheptene-1-carboxylic acid and cyclooctanecarboxylic acid accelerated epileptiform activities only slightly or were without effects. (iii) The small VPA-ester, 2-propylpentanoic acid ethyl ester, decreased the epileptiform activities in a way that is comparable to the effects of VPA well known from previous studies. It thus could be thought as a VPA-pro-drug. (iv) The mannitol-esters 1-O-(2-propylpentanoyl)-D-mannitol and 3,4;5,6-Di-O-isopropylidene-1-O-(2-propylpentanoyl)-D-mannitol blocked the PDS in a way which is different from the known effects of VPA. These substances are interpreted not to exert their effects after being metabolized to VPA and thus they are thought to be new antiepileptic substances. PMID- 10670422 TI - Effects of valproate derivatives II. Antiepileptic efficacy in relation to chemical structures of valproate sugar esters. AB - The structure effect relationships of derivatives of the antiepileptically active ester of valproate (VPA) 3,4:5,6-Di-O-isopropylidene-1-O-(2-propylpentanoyl)-D mannitol (1) have been studied using intracellular recording to record the membrane potential of single neurons (buccal ganglia, Helix pomatia). Epileptiform activity was induced by the epileptogenic drug pentylenetetrazol. The effects of several derivatives on epileptiform activity were compared with those of the relay compound 1. Most of the synthesized agents decreased the duration of paroxysmal depolarization shifts (PDS) and increased their repetition rate. It was considered that a decreased the duration of PDS is antiepileptic and an increased repetition rate is pro-epileptic. Compared with the effects of compound 1, the following relationships were found: (1) Derivatives containing glucitol or galactitol were of similar antiepileptic potency. (2) Introduction of pyranoses or furanoses rendered the substances inactive or even pro-epileptic. (3) VPA in position 1 and 6 at the sugar acted as an antiepileptic whereas in position 3 and 4 it proved to be ineffective. (4) Replacement of VPA by ethylhexanoyl reduced the antiepileptic potency slightly and pivaloyl strongly. (5) Replacement of isopropylidene bridges by penta-O-acetyl or cyclohexylidene residues led to largely inactive substances. (6) Compounds having isopropylidene bridges in position 2,4;3,5 proved to be antiepileptic whereas bridges especially in positions 2,3:4,5 slightly enhanced epileptic activities. PMID- 10670423 TI - Agonist-stimulated [35S]GTPgammaS binding in brain modulation by endogenous adenosine. AB - Coupling of receptors to G-proteins can be assessed by the ability of specific agonists to stimulate [35S]GTPgammaS binding in both brain membranes and sections in the presence of excess GDP. In some brain regions, however, high basal activity makes it difficult to detect agonist-stimulated [35S]GTPgammaS binding. The present study suggests a modification of the assay to reduce basal [35S]GTPgammaS binding and thus increase the signal:noise ratio. Adenosine A1 receptors belong to the class of G-protein-coupled receptors that activate Gi/Go proteins in brain. In the present study, the A1 agonist R(-)N6-(2 phenylisopropyl)adenosine (R-PIA) stimulated [35S]GTPgammaS binding in brain regions known to contain A1 receptors, including cerebellum, hippocampus and dentate gyrus, medial geniculate body, superior colliculus, certain thalamic nuclei, cerebral cortex, piriform cortex, caudate-putamen, and nucleus accumbens. Treatment of sections and membranes with adenosine deaminase (ADase), which is typically used in adenosine assays to eliminate endogenous adenosine, reduced basal [35S]GTPgammaS binding. In addition, for cannabinoid and mu-opioid agonists, the percent stimulation of [35S]GTPgammaS binding was approximately doubled when ADase was included in the assay. These results suggest that endogenous adenosine contributes significantly to basal [35S]GTPgammaS binding in certain brain regions, and that this activity may be reduced by the addition of ADase, thus improving the signal:noise ratio of agonist-stimulated [35S]GTPgammaS binding. PMID- 10670424 TI - Nicotine-evoked [3H]5-hydroxytryptamine release from rat striatal synaptosomes. AB - The aim of this study was to characterize the pharmacology of presynaptic nicotinic cholinoceptors (nAChRs) that modulate release of 5-hydroxytryptamine (5 HT) from superfused rat brain synaptosomes preloaded with [3H]5-HT. Nicotine increased 5-HT release from striatal synaptosomes (maximally by 15-30%) but not from cerebral cortex or hippocampal synaptosomes. Release of striatal 5-HT was increased in a concentration-dependent manner by nicotine, epibatidine, cytisine, and ACh (with added esterase inhibitor and muscarinic antagonist). Respective EC50 values were: 0.5, 0.003, 0.1 and 0.7 microM. The maximal effect of each agonist was virtually completely blocked by a high concentration of the insurmountable nicotinic antagonist mecamylamine; at a higher concentration of epibatidine (3 microM), a mecamylamine-insensitive effect was revealed. Nicotine, ACh and epibatidine appeared equally efficacious, whereas cytisine was of lower efficacy (60-70% of ACh). Release evoked by a half-maximal concentration of nicotine was inhibited by the nicotinic antagonists dihydro-beta-erythroidine (IC50 0.04 microM) and methyllycaconitine (IC50 0.06 microM). Nicotine-evoked 5 HT release was not reduced by tetrodotoxin given in a concentration that blocked veratridine-evoked release. These findings provide functional evidence for a direct action of nicotine on 5-HT neurons in the brain. The presynaptic nAChRs that modulate striatal 5-HT release appear to possess a novel pharmacological profile. PMID- 10670425 TI - Anxiogenic effects of nicotine in the dorsal hippocampus are mediated by 5-HT1A and not by muscarinic M1 receptors. AB - After direct administration into the dorsal hippocampus nicotine decreased the time spent in social interaction, without changing locomotor activity, indicating an anxiogenic effect. The possibility that post-synaptic M1 muscarinic receptors mediated this effect was examined by determining whether dorsal hippocampal administration of a specific M1 receptor agonist (McN-A-343) had anxiogenic effects, and whether the anxiogenic effect of nicotine could be reversed by co administration of the M1 receptor antagonist, pirenzepine. McN-A-343 (0.3, 1.6, 3.2, 15.8 nmol) was without effect on social interaction, and pirenzepine (0.7 and 2.4 nmol) injection into the dorsal hippocampus failed to reverse the decrease in social interaction caused by nicotine (6.3 nmol) injection into this area. However, the decrease in social interaction after nicotine (50 nmol) was completely reversed by the specific 5-HT1A receptor antagonist, WAY 100635 (0.4 nmol) after co-administration of both drugs into the dorsal hippocampus. Thus, the anxiogenic effect of nicotine in this brain region seems to be mediated by 5 HT1A, but not M1, receptors. In contrast to the effect of nicotine in naive animals, those retested after a second injection of 50 nmol did not show a significant anxiogenic effect. The theoretical implications of this are discussed and from a practical point of view this suggests caution in the retesting of animals after central injections. PMID- 10670426 TI - Exon-intron organization of the human 5-HT3A receptor gene. AB - The gene structure of the human 5-HT3A receptor gene was analyzed by exon to exon polymerase chain reaction and subsequent sequencing. The results were confirmed by restriction analysis and genomic Southern blotting. The coding region of the human gene was found to be split by eight introns at identical positions as in the murine 5-HT3A receptor gene. All exon-intron boundaries exhibited fully conserved splice donor and acceptor consensus sequences. The alternative splice acceptor in intron eight of the murine gene was not found in the human counterpart. The length of particular introns differs markedly from the murine gene. With the exception of intron 5, all human introns are longer than their murine counterparts. From the start to the stop codon the human gene stretches over about 14.5 kb. The human exon sequences confirm one of three published human 5-HT3A receptor cDNA sequences. Knowledge of the gene structure, including 1.9 kb of the 5' noncoding region, all introns and the exon-intron boundaries of the human 5-HT3A receptor gene should facilitate investigation of its potential role in psychiatric disorders. PMID- 10670427 TI - The effect of the NK1 receptor antagonist CP-99,994 on emesis and c-fos protein induction by loperamide in the ferret. AB - The site of the anti-emetic action of the neurokinin1 receptor antagonist CP 99,994 was studied in the ferret using the centrally acting opiate receptor agonist loperamide at a dose (0.5 mg/kg s.c.) which induced emesis in all animals tested. CP-99,994 (1 mg/kg, s.c.x2) abolished the emetic response (retching and vomiting) and the behaviours (licking, wet dog shakes, mouth scratching and gagging) induced by loperamide over a 2-h observation period. The enantiomer of this compound CP-100,263 (1 mg/kg, s.c.x2) did not have any significant effect on emesis or related behaviours. Loperamide (0.5 mg/kg s.c.) administration (but not its vehicle) resulted in dense fos-like immunoreactivity (FLI) mainly throughout the rostro-caudal extent of the nucleus tractus solitarius but not the area postrema. Although CP-99,994 (1 mg/kgx2) abolished the loperamide-induced emesis, it did not have any statistically significant effect on FLI in the brainstem. In loperamide and CP-100,263 (1 mg/kg, s.c.x2) treated animals FLI was comparable to that in animals treated with loperamide and CP-99,994. The results from this study taken together with those from previous studies indicate that loperamide exerts its emetic effect via nucleus tractus solitarius dendrites projecting into the area postrema. The lack of significant effect of CP-99,994 on the FLI induced by loperamide in this nucleus suggests that it is acting at a site "deep" in the nucleus tractus solitarius or elsewhere. The marked reduction in behaviours associated with loperamide administration by CP-99,994 provides a preliminary indication that NK1 receptor antagonist (as represented by CP-99,994) may in the clinic have effects on behaviours induced by emetic agents in addition to their previously described effects on retching and vomiting. PMID- 10670428 TI - Rolipram suppresses experimental autoimmune neuritis and prevents relapses in Lewis rats. AB - Rolipram, a phosphodiesterase type 4 inhibitor, can markedly down-regulate antigen-driven T cell proliferation and suppress TNF-alpha production in vitro and in vivo. Here we report the effects of Rolipram on experimental autoimmune neuritis (EAN), which can be induced by immunization with myelin components of the peripheral nervous system (PNS) combined with Freund's complete adjuvant (FCA), and which represents a CD4+ T cell-mediated animal model for human Guillain-Barre syndrome. EAN induced in Lewis rats by inoculation with the PNS P2 protein peptide 57-81 and FCA was strongly suppressed by Rolipram administered twice daily intraperitoneally from day 9 post immunization (p.i.), i.e. after onset of clinical EAN. Suppression of EAN was associated with down-regulated myelin antigen-induced T cell responses as well as down-regulated IFN-gamma and TNF-alpha production. A relapse of clinical EAN occurred upon treatment of a short duration (7 days), while prolongation of treatment resulted in the prevention of clinical EAN relapse. There was no relationship between clinical EAN relapse and high levels of TNF-alpha. The immunomodulatory effects of Rolipram call for further research into the potential role of drugs acting on the immune system in the treatment of autoimmune diseases. PMID- 10670429 TI - A unique hormonal and behavioral hyporesponsivity to both forced novelty and d amphetamine in periadolescent mice. AB - The identification of critical ontogenetic periods of increased vulnerability to the effects of drugs of abuse could have a great psychobiological and clinical therapeutical importance. Potential age-related differences in the response of the hypothalamic-pituitary-adrenal (HPA) axis to both stress and psychostimulants has been tested here in an animal model of adolescence. Periadolescent (PND 33 43) and Adult (PND>60) mice of both sexes were injected with d-amphetamine (AMPH, 0, 2, or 10 mg/kg i.p.) and immediately faced with a mild psychological stress experience, i.e. placement in a novel environment. A detailed time-course analysis of both hormonal and behavioral profiles was performed, with animals being sacrificed for trunk-blood collection at different time-points during the test (before the injection, NT group; 15, 30, or 120 min after the injection). Basal corticosterone (CORT) levels (NT group) were consistently higher in periadolescents than in adults. As a whole, a marked increment of blood CORT levels was found in mice of both ages exposed to forced novelty. However, important age-related differences were also observed, with Saline-injected periadolescents still exhibiting elevated levels of locomotion at the end of the 120-min test session and failing to show the increasing profile of CORT release over the baseline that was typical of adults. Upon an AMPH 2 administration, periadolescents exhibited a much lower profile of locomotor hyperactivity than adults, and also failed to show an increase across the course of the session in CORT release, that was observed in adults. When treated with the high AMPH 10 dose, a marked locomotor hyperactivity was found in periadolescents, which however showed much lower levels of the stereotyped licking and gnawing behavior, that was typical of adults. The present results suggest a unique profile of integrated behavioral and physiological hyporesponsivity in mice during periadolescence. The latter also represents a very useful model for the study of the issue of psychobiological risk factors involved in vulnerability to drugs of abuse in human adolescents. PMID- 10670430 TI - Involvement of tyrosine kinase in the pyrogenic fever exerted by NOS pathways in organum vasculosum laminae terminalis. AB - Nitric oxide synthase (NOS) is an enzyme which has a distinct cytokine-inducible isoform (iNOS). Many cytokine receptors have an intracellular tyrosine kinase domain. Here we have used two tyrosine kinase inhibitors (genistein and lavendustin A) to investigate the potential role of tyrosine kinase activation in the induction on both iNOS and fever caused by lipopolysaccharide (LPS) in rabbits. Direct administration of LPS into the organum vasculosum laminae terminalis (OVLT) increased iNOS expression. These increases paralleled the increase in deep body temperature in unanesthetized rabbits. Pretreatment with genistein or lavendustin A not only reduced the fever but also attenuated the iNOS expression in the OVLT following an intra-OVLT dose of LPS. These results suggest that tyrosine phosphorylation is part of the signal transduction mechanism that mediates the induction of both iNOS and fever elicited by LPS in the OVLT of rabbit brain. PMID- 10670431 TI - Developmental neuroplasticity in a model of cerebral hemispherectomy and stroke. AB - Cerebral hemispherectomy, a last resort treatment for childhood epilepsy, is a standard procedure which dramatically illustrates the resilience of the brain to extensive damage. If this operation, also mimicking long-term, extensive unilateral capsular stroke, is performed in postnatal cats of up to 60 days of age, there is a remarkable recovery/sparing of neurological functions that is not seen when the lesion occurs during late fetal life or in adulthood. A long-term effect at all ages is loss of neurons in bilateral brain areas remote from the resection site. This is pronounced in adult cats and shows intriguing, paradoxical features in fetal animals, but is substantially attenuated in neonatal cats. Similarly, large-scale reinnervation of subcortical sites (sprouting) by neurons of the remaining, intact hemisphere is prominent in young cats, but not in fetal or adult animals. These and other restorative processes (described herein) in young postnatal animals are matched by relatively higher rates of local cerebral glucose utilization, supporting the notion that they underlie the improved behavioral outcome. Thus, during a critical, defined stage of maturation, presumably common to higher mammals including humans, the brain entirely remodels itself in response to extensive but focal injury. Perhaps the molecular environment allowing for rescue of neurons and enhanced reinnervation at a specific developmental stage could be recreated in subjects with brain lesions at less favorable ages, thereby helping to restore circuitry and spare neurons. However, replacement via transplantation of neurons eliminated by the damage appears to be crucial in attempts to further preserve cells located remotely but yet destined to die or decrease in size. This article presents abundant evidence to show that there is a surprisingly comprehensive long-term morphological remodeling of the entire brain after extensive unilateral damage and that this occurs preferentially during a discrete period of early life. Additional evidence strongly suggests that the remodeling underlies the outstanding behavioral and functional recovery/sparing following early cerebral hemispherectomy. We argue that this period of reduced brain vulnerability to injury also exists in other higher mammals, including man, and suggest ways to enhance restorative processes after stroke/hemispherectomy occurring at other ages. PMID- 10670432 TI - Impaired neurotransmitter release and elevated threshold for cortical spreading depression in mice with mutations in the alpha1A subunit of P/Q type calcium channels. AB - The P/Q type voltage-gated Ca2+ channels are involved in membrane excitability and Ca2+-dependent neurotransmitter release within the CNS. Mutations in the CacnalA gene encoding the alpha1A subunit of the P/Q type Ca2+ channel have recently been reported in tottering mice and a more severely affected allele, leaner. Here we show using in vivo cortical microdialysis that evoked increases of extracellular glutamate levels are markedly attenuated in both mutants upon KCl-induced depolarization compared with wild-type mice. Tottering and leaner mice also show a 10-fold resistance to cortical spreading depression induced by cortical electrical stimulation or KCl application to the pial surface. A slower transcortical propagation speed and failure to sustain regenerative spread of the depolarizing wave were more pronounced in leaner neocortex. Both signaling defects appeared unrelated to the developmental history of repeated cortical spike-wave discharges, since neither were observed in the stargazer mouse, a Ca2+ channel gamma2 subunit mutant with a similar seizure phenotype. These data demonstrate two cortical excitability defects revealed by prolonged depolarization in cerebral networks expressing mutant P/Q type Ca2+ channels, and are the first to identify a gene linked to a spreading depression phenotype. PMID- 10670433 TI - The medial prefrontal cortex plays an important role in the excitation of A10 dopaminergic neurons following intravenous muscimol administration. AB - Intravenous muscimol administration increases the activity of dopaminergic neurons of the A10 cell group, located in the ventral tegmental area. Evidence suggests that this increase in activity is produced by disinhibition following the inhibition of GABAergic ("non-dopaminergic") cells in the ventral tegmental area. We hypothesized that the activation of A10 cells by muscimol is likely to be at least partly caused by the action of excitatory afferents. To verify this, A10 cells were isolated from ipsilateral afferent sources which utilise excitatory amino acids (which play an important role in the activity of these neurons), using hemisections at the level of the subthalamic nucleus (or just anterior to the subthalamic nucleus), electrolytic lesions of the pedunculopontine tegmental nucleus, or a combination of both. Following hemisections, and hemisections combined with lesions of the pedunculopontine tegmental nucleus, muscimol inhibited rather than excited A10 dopaminergic neurons. The pedunculopontine tegmental nucleus itself appeared to make little intrinsic contribution to muscimol-induced excitation, although the results suggested that part of the excitation which originates in the forebrain may be conducted to A10 cells via the pedunculopontine tegmental nucleus. The source of the effective forebrain excitation was investigated using electrolytic lesions of documented sources of excitatory amino acidergic afferents to the ventral tegmental area: the medial prefrontal cortex, certain nuclei of the amygdalar complex and the lateral habenular nucleus. In the medial prefrontal cortex lesioned group, muscimol again produced inhibition, an effect qualitatively and quantitatively similar to that in the hemisected groups. Habenular lesions blocked muscimol-induced excitation without producing inhibition, whilst amygdalar lesions produced no significant change in the effects of muscimol. The results suggest that under normal circumstances, an active excitation counteracts and exceeds the direct inhibitory effects of muscimol on the activity of A10 dopaminergic neurons. Furthermore, this activation appears to be produced by the action of excitatory (probably excitatory amino acidergic) afferents arising from the medial prefrontal cortex, and possibly the lateral habenular nucleus. Insofar as the excitation of A10 dopaminergic neurons, which is produced by certain drugs of abuse, and which may play a crucial role in their sustained use, has its basis in excitation following disinhibition, this excitation may provide a novel target for therapeutic intervention in addiction. PMID- 10670434 TI - Sensorimotor cortical influences on cuneate nucleus rhythmic activity in the anesthetized cat. AB - This work aimed to study whether the sensorimotor cerebral cortex spreads down its rhythmic patterns of activity to the dorsal column nuclei. Extracellular and intracellular recordings were obtained from the cuneate nucleus of chloralose anesthetized cats. From a total of 140 neurons tested (106 cuneolemniscal), 72 showed spontaneous rhythmic activity within the slow (< 1 Hz), delta (1-4 Hz), spindle (5-15 Hz) and higher frequencies, with seven cells having the delta rhythm coupled to slow oscillations. The spindle activity recorded in the cuneate was tightly coupled to the thalamo-cortico-thalamic spindle rhythmicity. Bilateral or contralateral removal of the frontoparietal cortex abolished the cuneate slow and spindle oscillations. Oscillatory paroxysmal activity generated by fast electrical stimulation (50-100 Hz/1-2 s) of the sensorimotor cortex induced burst firing synchronized with the paroxysmal cortical "spike" on all the non-lemniscal neurons, and inhibitory responses also coincident with the cortical paroxysmal "spike" in the majority (71%) of the cuneolemniscal cells. The remaining lemniscal-projecting neurons showed bursting activity (11%) or sequences of excitation-inhibition (18%) also time-locked to the cortical paroxysmal "spike". Additionally, the cerebral cortex induced coherent oscillatory activity between thalamic ventroposterolateral and cuneate neurons. Electrolytic lesion of the pyramidal tract abolished the cortically induced effects on the contralateral cuneate nucleus, as well as on the ipsilateral medial lemniscus. The results demonstrate that the sensorimotor cortex imposes its rhythmic patterns on the cuneate nucleus through the pyramidal tract, and that the corticocuneate network can generate normal and abnormal patterns of synchronized activity, such as delta waves, spindles and spike-and-wave complexes. The cuneate neurons, however, are able to generate oscillatory activity above 1 Hz in the absence of cortical input, which implies that the cerebral cortex probably imposes its rhythmicity on the cuneate by matching the intrinsic preferred oscillatory frequency of cuneate neurons. PMID- 10670435 TI - Cytotoxic lesion of the medial prefrontal cortex abolishes the partial reinforcement extinction effect, attenuates prepulse inhibition of the acoustic startle reflex and induces transient hyperlocomotion, while sparing spontaneous object recognition memory in the rat. AB - The partial reinforcement extinction effect refers to the increase in resistance to extinction of an operant response acquired under partial reinforcement relative to that acquired under continuous reinforcement. Prepulse inhibition of the acoustic startle response refers to the reduction in startle reactivity towards an intense acoustic pulse stimulus when it is shortly preceded by a weak prepulse stimulus. These two behavioural phenomena appear to be related to different forms of attentional processes. While the prepulse inhibition effect reflects an inherent early attentional gating mechanism, the partial reinforcement extinction effect is believed to involve the development of acquired inattention, i.e. the latter requires the animals to learn about what to and what not to attend. Impairments in prepulse inhibition and the partial reinforcement extinction effect have been independently linked to the neuropsychology of attentional dysfunctions seen in schizophrenia. The proposed neural substrates underlying these behaviourial phenomena also appear to overlap considerably: both focus on the nucleus accumbens and emphasize the functional importance of its limbic afferents, including that originating from the medial prefrontal cortex, on accumbal output/activity. The present study demonstrated that cytotoxic medial prefrontal cortex lesions which typically damaged the prelimbic, the infralimbic and the dorsal anterior cingulate areas could lead to the abolition of the partial reinforcement extinction effect and the attenuation of prepulse inhibition. The lesions also resulted in a transient elevation of spontaneous locomotor activity. In contrast, the same lesions spared performance in a spontaneous object recognition memory test, in which the lesioned animals displayed normal preference for a novel object when the novel object was presented in conjunction with a familiar object seen 10 min earlier within an open field arena. The present results lend support to the hypothesis that medial prefrontal cortex dysfunction might be related to some forms of attentional abnormality central to the symptomatology of schizophrenia. Relevance of the present findings in relation to the neural substrates underlying the partial reinforcement extinction effect and prepulse inhibition is further discussed. PMID- 10670436 TI - Orbital cortex neuronal responses during an odor-based conditioned associative task in rats. AB - Neuronal activity in the rat orbital cortex during discrimination of various odors [five volatile organic compounds (acetophenone, isoamyl acetate, cyclohexanone, p-cymene and 1,8-cineole), and food- and cosmetic-related odorants (black pepper, cheese, rose and perfume)] and other conditioned sensory stimuli (tones, light and air puff) was recorded and compared with behavioral responses to the same odors (black pepper, cheese, rose and perfume). In a neurophysiological study, the rats were trained to lick a spout that protruded close to its mouth to obtain sucrose or intracranial self-stimulation reward after presentation of conditioned stimuli. Of 150 orbital cortex neurons recorded during the task, 65 responded to one or more types of sensory stimuli. Of these, 73.8% (48/65) responded during presentation of an odor. Although the mean breadth of responsiveness (entropy) of the olfactory neurons based on the responses to five volatile organic compounds and air (control) was rather high (0.795), these stimuli were well discriminated in an odor space resulting from multidimensional scaling using Pearson's correlation coefficients between the stimuli. In a behavioral study, a rat was housed in an equilateral octagonal cage, with free access to food and choice among eight levers, four of which elicited only water (no odor, controls), and four of which elicited both water and one of four odors (black pepper, cheese, rose or perfume). Lever presses for each odor and control were counted. Distributions of these five stimuli (four odors and air) in an odor space derived from the multidimensional scaling using Pearson's correlation coefficients based on behavioral responses were very similar to those based on neuronal responses to the same five stimuli. Furthermore, Pearson's correlation coefficients between the same five stimuli based on the neuronal responses and those based on behavioral responses were significantly correlated. The results demonstrated a pivotal role of the rat orbital cortex in olfactory sensory processing and suggest that the orbital cortex is important in the manifestation of various motivated behaviors of the animals, including odor-guided motivational behaviors (odor preference). PMID- 10670437 TI - Identification of two persistently activated neurotrophin-regulated pathways in rat hippocampus. AB - Brain-derived neurotrophic factor contributes profoundly to modulate activity dependent synaptic plasticity in adult brain areas such as the hippocampus, but the mechanisms underlying this important role still remain unclear. Recently, we have shown that two serine/threonine kinases, calcium/calmodulin-dependent protein kinase-2 and casein kinase-2, are capable of mediating brain-derived neurotrophic factor responses in adult rat hippocampus. In the present study, using hippocampal slices from adult rat, we show that phospholipase C-regulated calcium signals couple the brain-derived neurotrophic factor receptor to two distinct pathways: a pathway in which calcium/calmodulin-dependent protein kinase 2 stimulates a signalling module involving the p38 subfamily of mitogen-activated protein kinases and its downstream target, usually named mitogen-activated protein kinase-activated protein kinase-2; and a pathway in which the extracellular signal-regulated kinase subfamily of mitogen-activated protein kinases activates casein kinase-2. Our results suggest that: (i) extracellular signal-regulated kinase is activated by B-Raf in response to a calcium-sensitive adenylate cyclase; and (ii) extracellular signal-regulated kinase activates casein kinase-2 via a protein phosphatase(s) that may be of the PP1 and/or PP2A type. Interestingly, we also show that neurotrophin-induced activation of the two signalling cascades promotes a sustained activation of mitogen-activated protein kinase-activated protein kinase-2 and casein kinase-2 in slices. Considering the ability of these two kinases to be persistently activated, and that most of the protein kinases which lie in these pathways are believed to be important for multiple events underlying neuronal plasticity, it is suggested that the mechanisms described here might contribute both to rapid synaptic changes through local effects and to long-lasting synaptic responses through new gene transcription in the hippocampus. PMID- 10670438 TI - The histological validation of post mortem magnetic resonance imaging-determined hippocampal volume in Alzheimer's disease. AB - For 11 AD cases and four normal elderly controls, post mortem volumes of the hippocampal subdivisions were calculated by using magnetic resonance imaging and histological sections. After at least six weeks of fixation in formalin, brains were examined on a 1.5-T Philips Gyroscan imager producing T1-weighted coronal images with a 3-mm slice thickness. Brains were then processed and embedded in paraffin. Serial coronal sections, 3 mm apart and stained with Cresyl Violet, were used for the planimetry and unbiased estimation of the total numbers of neurons in the hippocampal subdivisions. For all 15 cases, magnetic resonance imaging- and histology-based measurements were performed along the whole rostrocaudal extent of the hippocampal formation and included three subvolumes: (i) the hippocampus (CA1-CA4 and the dentate gyrus); (ii) hippocampus/subiculum; and (iii) hippocampus/parahippocampal gyrus. After controlling for shrinkage, strong correlations were found between magnetic resonance imaging and histological measurements for the hippocampus (r = 0.97, P < 0.001), hippocampus/subiculum (r = 0.95, P < 0.001) and hippocampus/parahippocampal gyrus (r = 0.89, P < 0.001). We also calculated the total number of neurons in the hippocampus and hippocampus/subiculum subvolumes. Strong correlations between the magnetic resonance imaging subvolumes and neuronal counts were found for the hippocampus (r = 0.90, P < 0.001) and the hippocampus/subiculum subvolume (r = 0.84, P < 0.001). We conclude that very accurate volumetric measurements of the whole hippocampal formation can be obtained by using a magnetic resonance imaging protocol. Moreover, the strong correlations between magnetic resonance imaging based hippocampal volumes and neuronal numbers suggest the anatomical validity of magnetic resonance imaging volume measurements. PMID- 10670439 TI - The effect of insulin and glucose on the plasma concentration of Alzheimer's amyloid precursor protein. AB - The deposition of beta amyloid is a critical event in the pathogenesis of Alzheimer's disease. This peptide is a metabolite of the amyloid precursor protein. Recent research suggests that there is a correlation between plasma insulin and glucose concentrations and memory performance in Alzheimer's disease sufferers. Additionally, in vitro evidence suggests that both insulin and glucose may affect the metabolism of amyloid precursor protein and therefore the production of beta amyloid--however, to our knowledge no in vivo data have yet been published. We investigated the effect of elevated plasma levels of glucose and insulin on the plasma concentration of amyloid precursor protein in non Alzheimer's disease subjects. As would be expected following ingestion of a glucose drink, blood insulin and glucose levels significantly increased. Interestingly, however, plasma amyloid precursor protein concentration decreased. Whilst no correlation was observed between insulin or glucose levels and plasma amyloid precursor protein concentration, the decrease in plasma amyloid precursor protein concentration was affected by the apolipoprotein E genotype of the subject. Possession of an epsilon4 allele resulted in a reduced decrease in plasma amyloid precursor protein in response to glucose ingestion when compared to non-epsilon4 subjects. We conclude that glucose ingestion, and the subsequent elevation of plasma levels of glucose and insulin leads to a decrease in plasma amyloid precursor protein concentration. Further studies are required to determine the clinical significance of these physiological changes in plasma amyloid precursor protein and the implications for Alzheimer's disease pathogenesis. PMID- 10670440 TI - Status epilepticus results in an N-methyl-D-aspartate receptor-dependent inhibition of Ca2+/calmodulin-dependent kinase II activity in the rat. AB - Status epilepticus is a major medical emergency that results in significant alteration of neuronal function. Status epilepticus involves seizure activity recurring frequently enough to induce a sustained alteration in brain function. This study was initiated to investigate how status epilepticus affects the activity of calcium and calmodulin-dependent kinase II in the brain. Calcium and calmodulin-dependent kinase II is a neuronally enriched signal transducing system involved in the regulation of neurotransmitter synthesis and release, cytoskeletal function, gene transcription, neurotransmitter receptor function and neuronal excitability. Therefore, alteration of this signal transduction system would have significant physiological effects. Status epilepticus was induced in rats by pilocarpine injection, allowed to progress for 60 min and terminated by repeated diazepam injections. Animals were killed at specific time-points and examined for calcium and calmodulin-dependent kinase II activity. Calcium and calmodulin-dependent kinase II activity was significantly reduced in cerebral cortex and hippocampal homogenates obtained from status epilepticus rats when compared with control animals. Once established, the status epilepticus-induced inhibition of calcium and calmodulin-dependent kinase II activity was observed at all time-points tested following the termination of seizure activity. However, calcium and calmodulin-dependent kinase II activity was not significantly decreased in thalamus and cerebellar homogenates. In addition, status epilepticus induced inhibition of calcium and calmodulin-dependent kinase II activity was dependent upon activation of N-methyl-D-aspartate subtype of glutamatergic receptors. Thus, status epilepticus induced a significant inhibition of calcium and calmodulin-dependent kinase II activity that involves N-methyl-D-aspartate receptor activation. The data support the hypothesis that inhibition of calcium and calmodulin-dependent kinase II activity may be involved in the alteration of neuronal function following status epilepticus. PMID- 10670441 TI - Ca2+ channels that activate Ca2+-dependent K+ currents in neostriatal neurons. AB - It is demonstrated that not all voltage-gated calcium channel types expressed in neostriatal projection neurons (L, N, P, Q and R) contribute equally to the activation of calcium-dependent potassium currents. Previous work made clear that different calcium channel types contribute with a similar amount of current to whole-cell calcium current in neostriatal neurons. It has also been shown that spiny neurons possess both "big" and "small" types of calcium-dependent potassium currents and that activation of such currents relies on calcium entry through voltage-gated calcium channels. In the present work it was investigated whether all calcium channel types equally activate calcium-dependent potassium currents. Thus, the action of organic calcium channel antagonists was investigated on the calcium-activated outward current. Transient potassium currents were reduced by 4 aminopyridine and sodium currents were blocked by tetrodotoxin. It was found that neither 30 nM omega-Agatoxin-TK, a blocker of P-type channels, nor 200 nM calciseptine or 5 microM nitrendipine, blockers of L-type channels, were able to significantly reduce the outward current. In contrast, 400 nM omega-Agatoxin-TK, which at this concentration is able to block Q-type channels, and 1 microM omega Conotoxin GVIA, a blocker of N-type channels, both reduced outward current by about 50%. These antagonists given together, or 500 nM omega-Conotoxin MVIIC, a blocker of N- and P/Q-type channels, reduced outward current by 70%. In addition, the N- and P/Q-type channel blockers preferentially reduce the afterhyperpolarization recorded intracellularly. The results show that calcium dependent potassium channels in neostriatal neurons are preferentially activated by calcium entry through N- and Q-type channels in these conditions. PMID- 10670442 TI - The immunophilin ligand FK506, but not GPI-1046, protects against neuronal death and inhibits c-Jun expression in the substantia nigra pars compacta following transection of the rat medial forebrain bundle. AB - The immunophilin ligand FK506 (Tacrolimus) is used for prevention of graft rejection following organ transplantation. FK506 is a high-affinity ligand for FK506-binding proteins, an immunophilin subgroup of peptidyl-prolyl-cis/trans rotamases abundant in the mammalian brain. Here, we demonstrate that FK506 is a potent survival factor that prevents neuronal cell death following axotomy of central intrinsic neurons. Administration of FK506 (2 mg/kg, s.c., per day for two days pre-axotomy and for up to eight days post-axotomy) effectively delayed and reduced the death of axotomized neurons in the substantia nigra pars compacta following transection of the medial forebrain bundle. In saline-treated controls, 75%, 89% and 92% of nigral neurons died after 25, 50 and 60 days post-axotomy, respectively. In contrast, application of FK506 resulted in survival of 46%, 44% and 28% of the axotomized nigral neurons, and the majority of these surviving neurons showed continuous expression of tyrosine hydroxylase, the pacemaker enzyme for dopamine synthesis. Moreover, FK506 significantly reduced the expression of the inducible transcription factor c-Jun and its N-terminal phosphorylation and prevented the axotomy-induced suppression of the constitutive transcription factor ATF-2 in neurons of the substantia nigra and mammillary body. The latter is also axotomized by the coincident transection of the mammillothalamic tract, but the mammillary neurons survive the axotomy. In contradistinction to FK506, the non-immunosuppressive FK506-binding protein ligand GPI-1046 (25 or 12.5 mg/kg, applied once or twice per day for two days pre axotomy and for eight days post-axotomy) was completely ineffective for all these parameters investigated. Finally, FK506, but not GPI-1046, impressively accelerated the recovery from surgery. Our data provide the first evidence that FK506 acts as a neuroprotective molecule that rescues axotomized otherwise degenerating central intrinsic neurons in the adult mammalian brain by mechanisms that interfere with the transcriptional program of the axotomy-induced cell body response, such as activating transcription factor-2 suppression and c-Jun expression and phosphorylation. PMID- 10670443 TI - Serotonergic transmission in the periaqueductal gray matter in relation to aversive behaviour: morphological evidence for direct modulatory effects on identified output neurons. AB - Intracellular recordings were made from 21 cells in the dorsolateral periaqueductal gray matter in coronal midbrain slices. In the majority (n = 20) bath application of 5-hydroxytryptamine (30 or 150 mM) evoked either hyperpolarizing (n = 11) or depolarizing (n = 9) responses. Reconstructions of 11 neurons in the dorsolateral periaqueductal gray matter after filling with biocytin revealed a population of output neurons whose axons followed a dorsolateral trajectory towards the perimeter of the ipsilateral periaqueductal gray matter. In seven cells, the axon could be followed into the adjacent mesencephalic reticular formation. At the light microscopic level, immunostaining for 5-hydroxytryptamine revealed immunoreactive processes throughout the dorsolateral periaqueductal gray matter but no labelled somata or dendrites. Close associations (i.e. no discernible gap) were observed between serotonergic profiles and the somata and dendrites of biocytin-filled cells. At the ultrastructural level, serial sections through 21 appositions on to biocytin filled dendrites in three slices revealed 19 true appositions (i.e. having closely parallel plasma membranes with no intervening glial cell profiles) with the biocytin-filled dendrite. Only four of the appositions (21%) showed evidence of synaptic specializations which included aggregations of synaptic vesicles, and some thickening of the apposing membrane. The dense reaction product in the biocytin-filled cells precluded identification of the ultrastructure of postsynaptic elements. However, examination of contacts between 5 hydroxytryptamine-immunoreactive profiles and unlabelled elements in material taken from the contralateral side of the periaqueductal gray matter (i.e. no biocytin present) or in material taken from perfusion-fixed whole brain, in which ultrastructural preservation was superior compared with slices, revealed a similar incidence (21% and 23%, respectively) of synaptic specializations. The data indicate that serotonergic transmission on to output neurons in the dorsolateral periaqueductal gray matter is largely mediated by non-junctional contacts, suggesting that the actions of 5-hydroxytryptamine on these cells are mediated predominantly by volume rather than wiring transmission. PMID- 10670444 TI - Activation of the parapyramidal region in the ventral medulla stimulates gastric acid secretion through vagal pathways in rats. AB - Neurons synthesizing thyrotropin-releasing hormone, substance P and serotonin in the medullary caudal raphe nuclei project to the dorsal vagal complex and play a role in the central vagal regulation of gastric function. Neurons in the parapyramidal region in the ventral medulla share similar biochemical coding and projections as those in the caudal raphe nuclei. The role of the parapyramidal region in the autonomic regulation of gastric acid secretion was investigated in urethane-anesthetized rats. Unilateral microinjection of kainate into the parapyramidal region at 10, 15 and 20 ng induced a dose-related stimulation of gastric acid secretion (net increases: 22.2+/-11.2, 40.5+/-8.5 and 89.8+/-19.4 micromol/60 min, respectively), while injection of vehicle had no effect (net change: -0.1+/-1.4 micromol/60 min). Time-course studies showed a nine-fold peak increase over basal at 30 min after parapyramidal injection of kainate (20 ng) and acid secretion returned to basal level at 70 min. Microinjections of kainate (15-20 ng) outside the parapyramidal region or into the parapyramidal region in vagotomized rats had no effect. Exposure to cold (4 degrees C) for 2 h, which is known to induce vagally mediated gastric secretory and motor responses through medullary thyrotropin-releasing hormone pathways, increased the number of Fos positive cells in the caudal, middle and rostral parts of the parapyramidal region to 4.3+/-0.4, 9.4+/-0.9 and 18.4+/-1.6/section, respectively, compared with 0.1+/-0. 1, 0.1+/-0.0 and 0.7+/-0.6/section, respectively, in rats maintained at room temperature. Most of the Fos-labeled cells co-expressed pro thyrotropin-releasing hormone messenger RNA signal and/or were serotonin immunoreactive. These data show that chemical activation of neurons in the parapyramidal region results in a vagal-dependent stimulation of gastric acid secretion and that acute cold exposure activates parapyramidal neurons containing pro-thyrotropin-releasing hormone and/or serotonin, suggesting a potential role of the parapyramidal region, in addition to the caudal raphe nuclei, as medullary sites involved in the vagal regulation of gastric function. PMID- 10670445 TI - Characterization of spinal amino acid release and touch-evoked allodynia produced by spinal glycine or GABA(A) receptor antagonist. AB - Intrathecal strychnine (glycine antagonist) or bicuculline (GABA(A) antagonist) yields a touch-evoked agitation that is blocked by N-methyl-D-aspartate receptor antagonism. We examined the effects of intrathecal strychnine and bicuculline on touch-evoked agitation and the spinal release of amino acids. Fifty-two Sprague Dawley rats were prepared under halothane anesthesia with a lumbar intrathecal catheter and a loop dialysis catheter. Four days after implantation, rats were randomized to receive an intrathecal injection of N-methyl-D-aspartate (3 microg), strychnine (3 microg) or bicuculline (10 microg), or a combination of N methyl-D-aspartate with bicuculline or strychnine. The agitation produced by brief light tactile stroking of the flank (tactile allodynia), and the spontaneous spinal release of glutamate, taurine and serine was measured. Intrathecal N-methyl-D-aspartate, strychnine and bicuculline produced similar touch-evoked allodynia. Intrathecal bicuculline and N-methyl-D-aspartate alone evoked a transient spinal release of glutamate and taurine, but not serine, in the 0- 10 min sample, while strychnine did not affect spinal transmitter release at any time. As GABA(A) but not glycine receptor inhibition at equi-allodynic doses increases glutamate release, while the allodynia of both is blocked by N methyl-D-aspartate receptor antagonism, we hypothesize that GABA(A) sites regulate presynaptic glutamate release, while glycine regulates the excitability of neurons postsynaptic to glutamatergic terminals. PMID- 10670446 TI - Evaluation of the activity of a novel metabotropic glutamate receptor antagonist (+/-)-2-amino-2-(3-cis and trans-carboxycyclobutyl-3-(9-thioxanthyl)propionic acid) in the in vitro neonatal spinal cord and in an in vivo pain model. AB - The cyclobutylglycine (+/-)-2-amino-2-(3-cis and trans-carboxycyclobutyl-3-(9 thioxanthyl)propionic acid) (LY393053) has been identified as a functionally potent metabotropic glutamate receptor antagonist. It is most potent on the two group I metabotropic glutamate receptors, 1alpha and 5alpha, with IC50 values of 1.0+/-0.4 microM and 1.6+/-1.4 microM, respectively. In this study, LY393053 has also been evaluated electrophysiologically on native group I metabotropic glutamate receptors in an in vitro spinal cord preparation as well as behaviourally, in a mouse model of visceral pain. LY393053 dose-dependently antagonised group I agonist, (RS)-3, 5-dihydroxyphenylglycine, or a broad spectrum agonist (1S,3R)-amino-1,3-cyclopentanedicarboxylic acid-induced depolarisation of spinal motoneurons. The apparent Kd values were estimated to be 0.3 microM against (RS)-3, 5-dihydroxyphenylglycine-induced depolarisation and 0.5 microM against (1S,3R)-amino-1,3-cyclopentanedicarboxylic acid-induced depolarisation, respectively. On the other hand, the dorsal root-ventral root potential elicited at 8 x threshold was depressed by LY393053 with IC50 values of 9.0+/-0.7 microM and 12.7+/-1.7 microM on monosynaptic and polysynaptic responses, respectively. When investigated using the mouse acetic acid writhing test, LY393053 showed significant analgesic effects at doses of 1-10 mg/kg intraperitoneally. An ED50 value of 6.0 mg/kg was obtained in this test. By revealing a potent effect of LY393053 in antagonising the native group I metabotropic receptor-mediated responses in the spinal cord in rodents, and an antinociceptive efficacy in a mouse visceral pain model, these results, therefore, provide additional evidence in support of the analgesic potential of metabotropic glutamate receptor antagonists. PMID- 10670447 TI - Sensory thresholds and the antinociceptive effects of GABA receptor agonists in mice lacking the beta3 subunit of the GABA(A) receptor. AB - A line of mice was recently created in which the gabrb3 gene, which encodes the beta3 subunit of the GABA(A) receptor, was inactivated by gene-targeting. The existence of mice with a significantly reduced population of GABA(A) receptors in the CNS enabled an investigation of the role of GABA and GABA(A) receptors in nociception. The present study examined the sensory thresholds of these mice, as well as the antinociceptive effects of subcutaneously or intrathecally administered GABA(A) and GABA(B) receptor agonists. Homozygous null (beta3-/-) mice displayed enhanced responsiveness to low-intensity thermal stimuli in the tail-flick and hot-plate test compared to C57BL/6J and 129/SvJ progenitor strain mice, and their wild-type (beta3+/+) and heterozygous (beta3+/-) littermates. The beta3-/- mice also exhibited enhanced responsiveness to innocuous tactile stimuli compared to C57BL/6J, 129/SvJ and to their beta3+/+ littermates as assessed by von Frey filaments. The presence of thermal hyperalgesia and tactile allodynia in beta3-/- mice is consistent with a loss of inhibition mediated by presynaptic and postsynaptic GABA(A) receptors in the spinal cord. As expected, subcutaneous administration of the GABA(A) receptor agonist 4,5,6,7-tetrahydroisoxazolo-(5,4 c)pyridin-3-ol did not produce antinociception in beta3-/- mice, whereas it produced a dose-dependent increase in hot-plate latency in C57BL/6J, 129/SvJ, beta3+/+ and beta3+/- mice. However, the antinociceptive effect of the GABA(B) receptor agonist baclofen in the tail-flick and hot-plate tests was also reduced in beta3-/- mice compared to the progenitor strains, beta3+/+ or beta3+/- mice after either subcutaneous or intrathecal administration. This finding was unexpected and suggests that a reduction in GABA(A) receptors can affect the production of antinociception by other analgesic drugs as well. PMID- 10670448 TI - Antinociceptive effects of the neuroactive steroid, 3alpha-hydroxy-5alpha-pregnan 20-one and progesterone in the land snail, Cepaea nemoralis. AB - Results of investigations with vertebrates have implicated neuroactive steroids and in particular 5alpha-reduced metabolites of progesterone such as 3alpha hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP/3A5P and originally allopregnanolone) in the rapid modulation of diverse functions including that of nociceptive sensitivity. These effects have been indicated to involve modulation of GABA receptors. Results of recent phylogenetic studies have revealed the presence of GABA receptors in invertebrates that may also be subject to modulation by steroids and neuroactive steroids. The present study examined the effects of the neuroactive steroid, 3alpha-hydroxy-5alpha-pregnan-20-one, as well as progesterone on aversive thermal (nociceptive) responses in a mollusc, the land snail, Cepaea nemoralis. 3alpha-Hydroxy-5alpha-pregnan-20-one had significant dose-related (0.01-1.0 microg) antinociceptive effects in Cepaea increasing the latency of response to a 40 degrees C surface, with maximum effects being evident 15-30 min after administration. These effects of 3alpha hydroxy-5alpha-pregnan-20-one were stereospecific, with the stereoisomer 3beta hydroxy-5alpha-pregnan-20-one (3B5P) failing to affect nociceptive responses. Progesterone also had significant dose-related (0.10-10 microg) antinociceptive effects that, however, were delayed in onset and relatively prolonged (60-120 min), suggestive of the formation of active metabolites. The presence of endogenous progesterone (12.36+/-0.17 ng/g tissue) was ascertained by a radioimmunoassay further supporting a functional role for steroids in Cepaea. The antinociceptive effects of 3alpha-hydroxy-5alpha-pregnan-20-one and progesterone were blocked by the GABA antagonists, bicuculline and picrotoxin, while being relatively insensitive to opioid and N-methyl-D-aspartate antagonists. These results suggest an early evolutionary development and phylogenetic continuity of neuroactive steroid and GABA involvement in the mediation of nociception. PMID- 10670449 TI - Tachykinin receptor inhibition and c-Fos expression in the rat brain following formalin-induced pain. AB - Recent pharmacological evidence has implicated substance P and neurokinin A, natural ligands for neurokinin-1 and neurokinin-2 receptors, respectively, as neurotransmitters in brain neuronal circuits activated upon noxious stimulation. The expression of the inducible transcription factor, c-Fos, was used to identify areas in the brain activated by a noxious stimulus (the subcutaneous injection of formalin), and to investigate the effects of intracerebroventricular administration of selective, nonpeptide antagonists for neurokinin-1 and neurokinin-2 tachykinin receptors on the neural activity in these areas and on the behavioural response to formalin-induced pain. Formalin (5%, 50 microl), injected subcutaneously through a chronically implanted catheter in the region of the lower hindlimb, increased c-Fos expression in a number of brain areas related to nociceptive transmission or the integration of stress responses. Grooming behaviour, licking and biting directed to the injected site, was the most frequent behavioural response. Intracerebroventricular pretreatment of rats with either RP 67580 (500 pmol), the active enantiomer of a neurokinin-1 receptor antagonist, or with SR 48968 (500 pmol), the active enantiomer of a neurokinin-2 receptor antagonist, reduced the formalin-induced c-Fos staining in the prefrontal cortex, dorsomedial and ventromedial nuclei of the hypothalamus, the locus coeruleus and the periaqueductal gray. The neurokinin-1, but not the neurokinin-2, receptor antagonist attenuated the formalin-induced activation of c Fos in the paraventricular nucleus of the hypothalamus. Simultaneous intracerebroventricular pretreatment with both neurokinin-1 and neurokinin-2 receptor antagonists did not produce any additional inhibitory effect on the post formalin c-Fos expression. None of the tachykinin receptor antagonists had an effect on the formalin-induced c-Fos expression in the septohypothalamic nucleus, medial thalamus, parabrachial nucleus and central amygdaloid nucleus, indicating that neurotransmitters other than neurokinins are most probably responsible for the activation of these areas in response to noxious stimulation. While both tachykinin receptor antagonists reduced the grooming behaviour to formalin, the neurokinin-1 receptor antagonist was clearly more effective than the neurokinin-2 receptor antagonist. Intracerebroventricular pretreatment of rats with the inactive enantiomers of the tachykinin receptor antagonists, RP 68651 and SR 48965, was without effect. Our results show that (i) the modified formalin test elicited an intense grooming behaviour and expression of c-Fos in numerous forebrain and brainstem areas, (ii) both tachykinin receptor antagonists were able to attenuate the behavioural response to pain and to reduce the formalin induced c-Fos expression in some, but not all, brain areas, and (iii) the neurokinin-1 antagonist, RP 67580, was more effective in inhibiting the behavioural response to formalin and the pain-induced activation of c-Fos than the antagonist for neurokinin-2 receptors, SR 48968, indicating that neurokinin-1 receptors are preferentially activated in neurokinin-containing pathways responding to noxious stimuli. Our results demonstrate that blockade of brain tachykinin receptors, especially of the neurokinin-1 receptor, reduces the behavioural response to pain and the pain-induced c-Fos activation in distinct brain areas which are intimately linked with nociceptive neurotransmission and the initiation and integration of central stress responses. Together with the previous findings of the inhibition of hypertensive and tachycardic responses to pain, the present data indicate that tachykinin receptor antagonists can effectively inhibit the generation of an integrated cardiovascular and behavioural response pattern to noxious stimuli. PMID- 10670450 TI - Heme oxygenase-1, heme oxygenase-2 and biliverdin reductase in peripheral ganglia from rat, expression and plasticity. AB - The expression of inducible and constitutive heme oxygenase and biliverdin reductase was studied in normal and cultured peripheral ganglia from adult rats, using immunocytochemistry and in situ hybridization. Dramatic changes were induced by one to two days' culturing of dorsal root ganglia, nodose ganglia, otic ganglia, sphenopalatine ganglia and superior cervical ganglia. An up regulation of inducible heme oxygenase was found in satellite cells of the cultured nodose ganglia, dorsal root ganglia, sphenopalatine ganglia and otic ganglia, whereas only a few satellite cells in the superior cervical ganglia responded with an increase in inducible heme oxygenase immunoreactivity. In the superior cervical ganglia inducible heme oxygenase also appeared in a subpopulation of macrophages. During culturing, expression of inducible heme oxygenase immunoreactivity also increased in axons and in nerve cell bodies. In situ hybridization corroborated the immunocytochemical findings, revealing a strong up-regulation of inducible heme oxygenase messenger RNA in satellite cells, and less pronounced up-regulation in nerve cell bodies. Constitutive heme oxygenase immunoreactivity was found in most neurons in all of the ganglia studied. No significant changes in constitutive heme oxygenase immunoreactivity could be observed in cultured ganglia. Biliverdin reductase immunoreactivity was barely detectable in any of the normal ganglia; however, after culturing it appeared in axons, single nerve cell bodies and nerve cell nuclei. The results show that inducible heme oxygenase is up-regulated in peripheral ganglia after axonal injury, and suggest a role for carbon monoxide in cellular signaling and a requirement for the antioxidant (bilirubin) during the regeneration process. PMID- 10670451 TI - Selective neuroprotective effects with insulin-like growth factor-1 in phenotypic striatal neurons following ischemic brain injury in fetal sheep. AB - Severe perinatal asphyxia can lead to injury and dysfunction of the basal ganglia. Post insult administration of insulin-like growth factor-1 is neuroprotective, particularly in the striatum. Insulin-like growth factor-1 is also known to be a neuromodulator of several types of striatal neurons. The striatum comprises various phenotypic neurons with a complex neurochemical anatomy and physiology. In the present study, we examined the specificity of neuronal rescue with insulin-like growth factor-1 on different striatal neurons. Bilateral brain injury was induced in near term fetal sheep by 30 min of reversible carotid artery occlusion. A single dose of 3 microg of insulin-like growth factor-1 was infused over 1 h into the lateral ventricle 90 min following ischemia. The histological and immunohistochemical outcome were examined after 4 days recovery using paraffin tissue preparations. Insulin-like growth factor-1 treatment (n = 11) significantly reduced the percentage of neuronal loss in the striatum compared with the vehicle treated group (n = 10, 28.3+/-5.1% vs 55.5+/ 17.3%, P < 0.005). Immunohistochemical studies showed that ischemia resulted in a significant loss of calbindin-28kd, choline acetyltransferase, parvalbumin, glutamate acid decarboxylase, neuronal nitric oxide synthase and neuropeptide Y immunopositive neurons, compared with sham controls. Insulin-like growth factor-1 markedly prevented the loss of calbindin-28kd (n = 7, P < 0.05), choline acetyltransferase (n = 7, P < 0.05), neuropeptide Y (n = 7, P < 0.05), neuronal nitric oxide synthase (n = 8, P < 0.05) and glutamate acid decarboxylase (n = 9, P < 0.05) immunopositive neurons, but failed to protect parvalbumin (n = 6) immunopositive neurons. The present study indicates that the therapeutic effect of insulin-like growth factor-1 in the basal ganglia is selectively associated with cholinergic and some phenotypic GABAergic neurons. These data suggest a potential role for insulin-like growth factor-1 in preventing cerebral palsy due to perinatal asphyxia. PMID- 10670452 TI - Distribution of glutamate and preproenkephalin messenger RNAs following transient focal cerebral ischemia. AB - Middle cerebral artery occlusion may result in increased activation of N-methyl-D aspartate- or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-type receptors by glutamate and lead to neuronal cell death. To characterize molecular events that precede cell death following transient focal ischemia, in situ hybridization histochemistry was used to measure levels of glutamate receptor subunit 1 (GluR1), GluR2, GluR3, N-methyl-D-aspartate receptor subunit 1 (NR1) and preproenkephalin messenger RNAs in adult rats at various recirculation times (1.5, 3 and 24 h) following a 90-min period of middle cerebral artery occlusion. At 1.5 and 3 h recirculation, autoradiography showed pronounced but differential decreases in AMPA, NR1 and preproenkephalin messenger RNA expression throughout the infarcted ipsilateral striatum. Non-uniform patterns of in situ hybridization grains emerged such that many striatal neurons were depleted of AMPA and preproenkephalin messenger RNAs, while others retained control levels. In cortical regions destined to undergo infarction, GluR2 and NR1 messenger RNAs were preferentially reduced relative to the contralateral side (to 75+/-8.5% and 66+/-4.5%, respectively); GluR1, GluR3 and preproenkephalin messenger RNAs were unaltered. At 24 h recirculation, depletion of striatal and cortical messenger RNAs became less selective. GluR3 and preproenkephalin messenger RNAs were up regulated in ipsilateral spared regions of the striatum, and GluR1 and GluR2 messenger RNAs increased bilaterally in the cingulate cortex and in selective nuclei of the amygdala. Histological cell death or neurodegeneration was not detected in areas of reduced glutamate and preproenkephalin messenger RNA expression in either the ipsilateral striatum or cortex before 24 h. These findings suggest that complex and long-lasting decreases in messenger RNA expression occur prior to significant cell loss in regions destined to undergo infarction. Increased formation of Ca2+-permeable AMPA receptor assemblies may occur in "unspared" and "spared" regions via different mechanisms and contribute to alterations in post-ischemic synaptic activity. The possibility arises that there may be altered relationships between glutamatergic and enkephalin synapses, since the dorsolateral striatum, where preproenkephalin messenger RNA expression is acutely reduced, receives innervation by the affected ipsilateral cortical region. PMID- 10670454 TI - Patterns of developmental expression of the RNA editing enzyme rADAR2. AB - To date, two structurally related RNA-editing enzymes with adenosine deaminase activity have been identified in mammalian tissue: ADAR1 and ADAR2 [Bass B. I. et al. (1997) RNA 3, 947-949]. In rodents, ADAR2 undergoes alternative RNA splicing, giving rise to two splice variants that differ by the presence or absence of a 10 amino-acid insert in the carboxy-terminal catalytic domain. However, the physiological significance of the splicing and its regional and developmental regulation are as yet unknown. The present study examined spatial and temporal patterns of ADAR2 gene transcripts within specific neuronal populations of rat brain. The two rodent ADAR2 isoforms were expressed at comparable levels at all ages examined. rADAR2 messenger RNA expression was first detectable in the thalamic nuclei formation at embryonic day E19. The rADAR2b insert and rADAR2a splice probes produced images similar to that of the rADAR2 pan probe. At birth, rADAR2a messenger RNA splice variants were abundantly expressed in the thalamic nuclei. No signal for any probe was detectable in other brain regions, including neocortex, hippocampus, striatum and cerebellum at this stage of development. During the first week of postnatal life, rADAR2 messenger RNA expression (detected with the pan probe) increased gradually in several brain regions, with low expression detected at postnatal day P7 in the olfactory bulb, inferior colliculus, and within the pyramidal and granule cell layers of the hippocampus. Hybridization patterns of the rADAR2a variant probe reached peak expression at about the second week of life, while peak expression of the rADAR2b probe was reached at about the third week of life. At the end of the first week of life (P7), expression of both splice variants was strongest in the thalamic nuclei. By P14, rADAR2 messenger RNA expression was more consolidated in the deeper structures, including the thalamic nuclei and the granule cell layer of the cerebellum. By P21, maximal levels of rADARb expression were observed in the thalamic nuclei, inferior colliculus, cerebellum and pontine nuclei. In the adult, rADAR2 messenger RNA expression was of highest intensity in the thalamic nuclei, with high levels of expression in the olfactory bulb, inferior colliculus, cerebellum and pontine nuclei. At the level of the hippocampus, positive labelling was restricted to the CA3 region of the Ammon's horn and the dentate gyrus, with weak signals in the CA1 subfield. rADAR2 pan expression was at near background levels throughout the neocortex and caudate putamen. In summary, our study shows that ADAR2 messenger RNA expression is regulated in a cell-specific manner throughout development. At early ages, ADAR2 messenger RNA is expressed only within (and restricted to) the thalamic nuclei. By the third postnatal week, expression of the editase enzyme is more widely distributed throughout the olfactory bulb, CA3 and dentate gyrus of the hippocampus, thalamus, inferior colliculus and the molecular cell layer of the cerebellum. ADAR2 is thought to act at specific nucleotide positions in primary transcripts encoding glutamate receptor subunits, thereby altering gating and ionic permeability properties of AMPA- and kainate-activated channels. ADAR2 also acts at pre-messenger RNA encoding the serotonin 5HT-2C receptor to alter G-protein coupling. Thus, RNA editing may be an important mechanism for fine-tuning of the physiological and pharmacological properties of transmitter receptors of the central nervous system. PMID- 10670453 TI - Induction of gap junctional intercellular communication, connexin43 expression, and subsequent differentiation in human fetal neuronal cells by stimulation of the cyclic AMP pathway. AB - Expression of gap junction proteins and cell-cell communication was studied in the human neural-glial cell line, SVG, as a first step in defining whether the SVG cells could be used as a model system to study the role of gap junctions in neuronal precursor cells. SVG cells were found to express connexin43 protein that co-migrated with WB-F344 rat liver connexin43 and that reacted with connexin43 specific antibodies on Western blots. However, fluorescence recovery after photobleaching analysis of 5,6-carboxyfluorescein-loaded cells failed to show significant dye coupling. Agents that stimulate the adenylyl cyclase/cAMP pathway were used to induce gap junctional intercellular communication in the SVG cultures. A 24-48 h treatment of SVG cells with 5 microM forskolin or 5 microM forskolin + 200 microM 3-isobutyl-1-methylxanthine increased the percentage of dye-coupled cells from 5-65%, using the fluorescent recovery after photobleaching method. The increase in dye coupling induced by forskolin or forskolin + 3 isobutyl-1-methylxanthine was inhibited by octanol, which is known to block gap junction-mediated cell communication. Western blot analysis of total protein extracts revealed the appearance of a higher molecular weight connexin43 protein band after treatment of SVG cells with forskolin or forskolin + 3-isobutyl-1 methylxanthine, that was not observed in vehicle-treated controls. Alkaline phosphatase treatment of total protein extracts from forskolin or forskolin + 3 isobutyl-1-methylxanthine-treated cells reduced the higher molecular weight band to approximately 41,000 the same as observed in the control extracts. The alkaline phosphatase treatment demonstrates that the higher molecular weight band was due to a phosphorylation event stimulated by forskolin or the forskolin + 3 isobutyl-1-methylxanthine combination. In addition, treatment of the SVG cells with the forskolin or forskolin + 3-isobutyl-l-methylxanthine stimulated outgrowth of neurite-like processes from the cell body which immunostained positive for the connexin43 protein as well as protein markers for neurons and oligodendrocytes. We hypothesize that the SVG cells may represent a neuronal progenitor cell population that has the ability to differentiate when exposed to the appropriate signals. PMID- 10670455 TI - Immunolabeling of the rat central nervous system with antibodies partially selective of the short form of the 5-HT3 receptor. AB - Polyclonal antibodies were raised against a synthetic hexadecapeptide corresponding to the portion of the second intracytoplasmic loop of the short form of the mouse 5-hydroxytryptamine-3A receptor subunit (5-HT3A-S), which differs from the long form (5-HT3A-L) by the removal of six amino acids. Antibodies were detected by enzyme-linked immunosorbent assay as soon as two months after the first injection to rabbits of the peptide coupled to keyhole limpet hemocyanin. Immunoblot detection of fusion proteins comprising glutathione S-transferase and the second intracellular loop of 5-HT3A-S or 5-HT3A-L, and immunoprecipitation of cloned receptors showed that antibodies exhibited some selectivity for the short variant. Affinity chromatography allowed the purification of selective anti-5-HT3A-S antibodies which yielded a strong positive labeling of plasma membrane, reticulum and Golgi apparatus of COS-7 cells expressing murine 5-HT3A-S. In contrast, COS-7 cells expressing similar levels of 5-HT3A-L exhibited only a very weak labeling. Selectivity was also observed on immunoblots of cloned receptors transiently expressed in COS-7 cells, or stably expressed in CHO cells, both systems showing an immunolabeled component at 53,000-54,000 mol. wt. Immunoautoradiographic labeling of central nervous system sections showed that 5-HT3A-S-like immunoreactivity was found mostly within the nucleus of the solitary tract, the nucleus of the spinal tract of the trigeminal nerve, and the dorsal horn of the the spinal cord in the rat. After unilateral ablation of the nodose ganglion, 5-HT3A-S-like immunoreactivity decreased markedly in the ipsilateral part of the nucleus of the solitary tract, as expected of the presynaptic localization of 5-HT3 receptors. Finally, immunohistochemistry at the light and electron microscope levels revealed that 5 HT3A-S-like immunoreactivity was associated essentially with terminals and axonal profiles. All these results demonstrate that the immunolabeling exhibited by these antibodies is consistent with a specific and partially selective recognition of the short isoform of the 5-HT3A subunit. Because the pattern of immunoautoradiographic labeling matches the distribution previously established with selective radioligands, it can be inferred that these antibodies probably recognized the same fully assembled form of the 5-HT3A-S receptor subunit. PMID- 10670456 TI - Immunolocalization of the arachidonic acid and mechanosensitive baseline traak potassium channel in the nervous system. AB - TRAAK is the sole member of the emerging class of 2P domain K+ channels to be exclusively expressed in neuronal cells. TRAAK produces baseline K+ currents which are strongly stimulated by arachidonic acid and by mechanical stretch, and which are insensitive to the classical K+ channel blockers tetraethylammonium, Ba2+, and Cs+. This report describes the immunolocalization of TRAAK in brain, spinal cord, and retina of the adult mouse. The most striking finding is the widespread distribution of the TRAAK immunoreactivity, with a prominent staining of the cerebellar cortex, neocortex, hippocampus, dentate gyrus, subiculum, the dorsal hippocampal commissure, thalamus, caudate-putamen, olfactory bulb, and several nuclei in the brainstem. Virtually all neurons express TRAAK, and the highest immunoreactivity was seen in soma, and to a lesser degree in axons and/or dendrites in most areas in brain and spinal cord. In the retina, the TRAAK protein is concentrated to the soma of ganglion cells and to the dendrites of all other neurons. Taken together, these results show a wide distribution of TRAAK, a mechanosensitive and arachidonic acid-stimulated neuron-specific baseline K+ channel, in brain, spinal cord and retina. PMID- 10670457 TI - Regional brain variations of cytochrome oxidase activity and motor co-ordination in staggerer mutant mice. AB - A mutant mouse with cerebellar cortical atrophy, staggerer, was examined in tests of motor activity and co-ordination as well as in regional brain metabolism as assessed by cytochrome oxidase activity. Compared with non-ataxic controls, staggerer mutants had inferior performances in the open field, the wooden beam, the wooden edge, and the rotorod tests. An increase in cytochrome oxidase activity in the deep cerebellar nuclei and in some cerebellar efferent regions, such as the lateral vestibular nucleus, the parvicellular red nucleus, and the ventral tegmental area, was found in staggerer mutant mice. Abnormally high cytochrome oxidase activity in the interpositus and the dentate nuclei was linearly correlated with poor performance on the wooden beam and on the rotorod. High cytochrome oxidase activity in the lateral vestibular nucleus was also associated with poor performance on the wooden beam. Moreover, high cytochrome oxidase activity in the fastigial nucleus was associated with poor performance on the wooden beam but with high motor activity in the open field. These results indicate that a lack of innervation of Purkinje cells to the deep cerebellar nuclei is in part the cause of motor co-ordination deficits in staggerer mutant mice. PMID- 10670458 TI - Separate roles for beta2- and beta3-adrenoceptors in memory consolidation. AB - Consolidation of a labile memory which would not normally be stored can be achieved by intracerebral administration of noradrenaline. In a series of experiments using discriminated, one trial passive avoidance learning with the day-old chick, the effect of noradrenaline has been shown to be due to actions at different subtypes of adrenoceptors. The effect of noradrenaline is dose dependent, with a moderate dose producing memory consolidation. However, higher doses of noradrenaline (0.3-10 nmol/hemisphere) prevent consolidation, an effect not seen with isoprenaline suggesting that these doses stimulate alpha adrenoceptors. The promotion of memory consolidation by noradrenaline or isoprenaline at low doses was attributable to beta3-adrenoceptors and at medium doses to beta2-adrenoceptors. At higher doses of noradrenaline, there was alpha1 adrenoceptor-mediated inhibition of memory consolidation. Consolidation can also be achieved by administration of either beta2- or beta3-adrenoceptor agonists at specific times after training. Although these two adrenoceptors both promoted memory consolidation, there was a differential action on the stages of memory formation. The dose-response curve to the beta3- and the beta2-agonists was shifted by the appropriate antagonist but not by the antagonist at the other beta adrenoceptor. Although beta1-adrenoceptors are present in chick brain, they do not seem to have a role in memory formation. These results explain why noradrenaline, acting at different adrenoceptors, can have different effects on memory formation with memory being either consolidated or inhibited depending on the dose. The findings also demonstrate a role in memory formation for beta3 adrenoceptors found in the brain. Agonists acting specifically at beta2- or beta3 adrenoceptors may be of value in diseases involving cognitive impairment. PMID- 10670459 TI - Long-term depolarization changes morphological parameters of PC12 cells. AB - It is well known that neuronal differentiation is strongly dependent on the intracellular level of free calcium ions ([Ca2+]i). In the present study the morphological and intracellular free calcium concentration changes were compared on PC12 pheochromocytoma cells cultured in control conditions and in a medium with high KCl level. Culturing PC12 cells in a medium with 20-30 mM KCl deprived of nerve growth factor supported cell proliferation and rapid growth of small neurite-like processes. However, their lengths did not increase with prolongation of the time of culturing. During culturing with 40 mM KCl the growth of these processes became blocked; the cells stopped proliferating and showed signs of degeneration. Measurements of [Ca2+]i level during the first days of PC12 cells culturing in a hyperpotassium medium indicate that such changes in this level could be an important factor in the induction of the observed morphological alterations; however, other effects induced by membrane depolarization may also be responsible for them. PMID- 10670460 TI - Biochemistry of visual pigment regeneration: the Friedenwald lecture. PMID- 10670461 TI - Self-destructive and self-protective processes in the damaged optic nerve: implications for glaucoma. PMID- 10670462 TI - Genes upregulated in the human trabecular meshwork in response to elevated intraocular pressure. AB - PURPOSE: To identify genes upregulated in perfused, intact human trabecular meshwork (TM) in response to elevated intraocular pressure (IOP). METHOD: Two pairs of anterior segments of normal human eyes from postmortem donors were placed in culture and perfused 24 hours at constant flow (3 microl/min). After reaching baseline, the flow of one eye from each pair was raised to obtain an incremental pressure (deltaP) of 50 mm Hg for 6 hours. The anterior segments were then quickly frozen in liquid nitrogen, and their TMs were dissected for RNA extraction. SMART cDNA libraries were generated from control and high-pressure human TM RNAs and hybridized to sets of identical high-density cDNA gene arrays. These arrays contained 18,376 human expressed sequence tags (ESTs), corresponding to both characterized and unknown genes. Differentially expressed genes were identified by different-intensity hybridization signals and confirmed by semi quantitative polymerase chain reaction. RESULTS: Eleven genes were found to be consistently upregulated in the human TM by elevated IOP: interleukin-6, preprotachykinin-1, secretogranin-II, cathepsin-L, stromelysin-1, thymosin-beta4, alpha-tubulin, alphaB-crystallin, glyceraldehyde-3-phosphate dehydrogenase, metallothionein and Cu/Zn superoxide dismutase. The products of these genes are involved in vascular permeability, secretion, extracellular matrix remodeling, cytoskeleton reorganization, and reactive oxygen species scavenging. CONCLUSIONS: Elevated IOP induced specific upregulation of 11 physiologically relevant genes. On the basis of their known activities, the products of each of these genes might predict homeostatic mechanisms similar to those involved in the regulation of blood vessel permeability. We hypothesize that similar mechanisms might be involved in regulating flow through Schlemm's Canal endothelium. PMID- 10670463 TI - Advanced glycation end products in diabetic corneas. AB - PURPOSE: Corneal complications are often associated with diabetes mellitus and can be vision threatening. Corneas in diabetic patients are exposed to increased glucose concentration despite cornea's avascular property, and this condition may contribute to the accumulation of advanced glycation end products (AGEs). The focus of this study was to examine the role of AGEs in the pathogenesis of diabetic keratopathy. METHODS: An anti-AGE monoclonal antibody (6D12), which recognizes a N(epsilon)-carboxymethyl lysine (CML)-protein adduct as an epitope, was prepared. Immunohistochemical localization of CML was examined in human age matched diabetic and nondiabetic corneas (8 of each). In vitro, type I collagen-, type IV collagen-, or laminin-coated 96-well plates were glycated by glucose phosphate. In some experiments, aminoguanidine was present in the incubation mixture. The amounts of CML-protein adducts in the extracellular matrix (ECM) were determined by enzyme-linked immunosorbent assay using 6D12. SV40 immortalized human corneal epithelial cells were seeded onto modified or unmodified ECM in 96-well plates and allowed to attach for 3 hours. Attached cells were fixed, and the areas of attached cells in each condition were measured. Attached cells without fixation were removed, and cell number was counted. RESULTS: In all of the 8 diabetic corneas, CML immunoreactivity was observed in the epithelial basement membrane, whereas CML immunoreactivity was not found in the corresponding area in 7 of 8 nondiabetic corneas. In vitro, nonenzymatic glycation of laminin on the culture dish attenuated adhesion and spreading of corneal epithelial cells. The presence of amninoguanidine in the incubation mixture during glycation inhibited CML formation and promoted the adhesion and spreading of corneal epithelial cells in a dose-dependent manner. CONCLUSIONS: The accumulation of AGEs on the basement membrane, particularly on laminin, may play a causative role in the corneal epithelial disorders of diabetic patients. PMID- 10670464 TI - Corneal stromal changes induced by myopic LASIK. AB - PURPOSE. Despite the rapidly growing popularity of laser in situ keratomileusis (LASIK) in correction of myopia, the tissue responses have not been thoroughly investigated. The aim was to characterize morphologic changes induced by myopic LASIK in human corneal stroma. METHODS: Sixty-two myopic eyes were examined once at 3 days to 2 years after LASIK using in vivo confocal microscopy for measurement of flap thickness, keratocyte response zones, and objective grading of haze. RESULTS: Confocal microscopy revealed corneal flap interface particles in 100% of eyes and microfolds at the Bowman's layer in 96.8%. The flaps were thinner (112 +/- 25 microm) than intended (160 microm). The keratocyte activation in the stromal bed was greatest on the third postoperative day. Patients with increased interface reflectivity due to abnormal extracellular matrix or activated keratocytes at > or = 1 month (n = 9) had significantly thinner flaps than patients with normal interface reflectivity (n = 18; 114 +/- 12 versus 132 +/- 22 microm, P = 0.027). After 6 months the mean density of the most anterior layer of flap keratocytes was decreased. CONCLUSIONS: Keratocyte activation induced by LASIK was of short duration compared with that reported after photorefractive keratectomy. The flaps were thinner than expected, and microfolds and interface particles were common complications. The new findings such as increased interface reflectivity associated with thin flaps and the apparent loss of keratocytes in the most anterior flap 6 months to 2 years after surgery may have important clinical relevance. PMID- 10670465 TI - Experimental corneal endotheliitis in rabbit. AB - PURPOSE: Corneal endotheliitis may cause permanent visual loss due to endothelial decompensation. The pathogenesis underlying this distinct clinical entity is not known. In the current study, a rabbit herpetic corneal endotheliitis model was made of induced anterior chamber-associated immune deviation (ACAID). METHODS: One group of rabbits received left-eye intracameral inoculation of UV-inactivated herpes simplex virus (HSV)-1 (strain McKrae). The second group received cell medium in the same manner as the first group. The third group subcutaneously received the same inoculum as the first group. Seven days later, all right eyes were intracamerally infected with 2.5 x 10(4) plaque-forming units of infectious HSV-1. Eyes were evaluated by slit lamp examination. Two weeks after infection, rabbits were killed, and right eyes were examined by immunohistochemical staining and electron microscopy. Aqueous humor was detected for HSV-1 DNA and antibody. RESULTS: Nonspecific inflammation occurred in the anterior segments of the eyes from the second and third groups. In contrast, at 14 days after infection, the first group of rabbits showed a specific pattern of inflammation that greatly resembled clinical features of corneal endotheliitis. Viral antigen was detected only in the endothelial layer. Electron microscopy revealed enlarged intercellular gaps and infiltration of inflammatory cells that are characteristic of endothelial defects. HSV-1 DNA was detected at a significantly higher number in the aqueous humor aspirates from endotheliitis rabbits. In addition, ACAID was shown to be induced in the rabbits with corneal endotheliitis. CONCLUSIONS: HSV-1 infection can induce corneal endotheliitis and ACAID may play the pivotal role in this entity. PMID- 10670466 TI - Cholinergic-induced Ca2+ elevation in rat lacrimal gland acini is negatively modulated by PKCdelta and PKCepsilon. AB - PURPOSE: To investigate the role of protein kinase C (PKC) in cholinergic agonist induced Ca2+ elevation in lacrimal gland acini. METHODS: Lacrimal gland acini were prepared by collagenase digestion, and changes in intracellular Ca2+ ([Ca2+]i) were measured using fura-2 as a fluorescent probe. RESULTS: Preactivation of PKC by phorbol 12-myristate 13-acetate (PMA), or inhibition of protein phosphatase type 1/2A (PP1/2A) by calyculin A, decreased both the [Ca2+]i transient and the plateau of [Ca2+]i induced by increasing concentrations of carbachol, a cholinergic agonist. Staurosporine, an inhibitor of PKC, completely reversed the effect of PMA. Inhibition of the Ca(2+)-independent PKC isoforms PKCdelta and -epsilon, but not the Ca(2+)-dependent isoform PKCalpha substantially reversed the inhibitory effect of PMA on cholinergic agonist induced Ca2+ elevation. The inhibitory effect of PMA was obtained only in the presence of extracellular Ca2+, suggesting that PKC inhibits the influx of Ca2+. PMA completely inhibited the cholinergic agonist-induced plateau of [Ca2+]i. PMA and calyculin A decreased both the [Ca2+]i transient and the plateau of [Ca2+]i induced by thapsigargin, further supporting the idea that PKC modulates the entry of Ca2+. CONCLUSIONS: In the lacrimal gland, agonist-induced changes in [Ca2+]i are negatively regulated by PKC-dependent phosphorylation of a target protein(s) that is sensitive to PP1/2A. PMID- 10670467 TI - Effect of myopic LASIK on corneal sensitivity and morphology of subbasal nerves. AB - PURPOSE: To investigate whether the morphology of the subbasal nerves corresponds to corneal sensitivity after laser in situ keratomileusis (LASIK). METHODS: In a case series study, 59 patients were examined at 2 to 4 hours, 3 days, 1 to 2 weeks, 1 to 2 months, 3 months, or 6 or more months after undergoing LASIK for myopia, by using a Cochet-Bonnet esthesiometer and an in vivo confocal microscope, and were compared with control subjects. Corneal sensitivity and confocal images of subbasal nerves were obtained centrally and 2 mm nasally and temporally. Subbasal nerve fiber bundles (NFBs) were grouped as follows: corneas with no nerve images; corneas with short (<200 microm), unconnected NFBs; corneas with long (> or =200 microm) NFBs without interconnections; and corneas with long NFBs with interconnections. RESULTS: Corneal sensitivity was at its lowest at 1 to 2 weeks after LASIK. Sensitivity of the hinge area was higher than temporal or central areas at every time point. At 6 or more months the sensitivity values were comparable with the values observed in control subjects. The central area showed mainly short, unconnected subbasal NFBs, even at 6 months. In general, the temporal area presented with long NFBs from 3 months onward, whereas the nasal area showed long NFBs at every time point. CONCLUSIONS: The results suggest that the corneal areas with no nerve images or short, unconnected NFBs are associated with lower sensitivities than corneal areas with long NFBs with or without interconnections. In vivo confocal microscopy reveals LASIK-induced alterations of subbasal nerve morphology and thus enables a direct comparison of corneal sensory innervation and sensitivity. PMID- 10670468 TI - Membrane-associated mucins in normal human conjunctiva. AB - PURPOSE: To examine the presence of specific membrane-associated mucins in normal human conjunctiva. METHODS: Glycoconjugates were extracted from membranes with two detergents: octylglucoside and Triton X114. Mucins were separated by cesium chloride density gradient centrifugation. Size was assessed by gel filtration on Sepharose CL2B and charge by ion-exchange chromatography on MonoQ. Cross reaction with antibodies against mucin gene products was assessed in blots of electrophoresis gels. RESULTS: Extraction of total tissue membranes yielded material with a buoyant density typical of mucins. Gel filtration showed material reacting with antimucin antibodies in a range of molecular sizes. Agarose electrophoresis confirmed the presence of MUC1 and MUC4 and the absence of MUC2 or MUC5AC. Isolation of membrane mucins by sequential, exhaustive extraction with octylglucoside followed by Triton X114 suggested the existence of mucins in different membrane environments. Reagents to carbohydrate epitopes revealed high mobility material, comigrating with MUC1 and MUC4. Low mobility membrane-bound mucins did not cross-react with any antibodies to mucin genes known to be expressed in human conjunctiva. CONCLUSIONS: Membrane-associated mucins are distinct from secreted mucins in normal human conjunctiva. MUC1 and MUC4 mature products decorate the membranes of conjunctival epithelial cells. Their segregation between octyl glucoside and the detergent and aqueous phases of Triton X114 suggests a variety of membrane anchoring modes. PMID- 10670470 TI - Motion VEPs, stereopsis, and bifoveal fusion in children with strabismus. AB - PURPOSE: The link between nasal-temporal motion asymmetries and anomalous binocular sensory function in infantile esotropia (ET) has led to the idea that visual evoked potential responses to horizontal motion (mVE) is an alternative measure of sensory binocularity to stereopsis. A second hypothesis is that the mVEP response is a marker for bifoveal fusion. The purpose of this study was to directly evaluate these two hypotheses by examining the correspondence between the mVEP response and both stereoacuity and bifoveal fusion in a cohort of strabismic patients with variable binocular sensory function. METHODS: Motion VEPs, random dot stereopsis, and bifoveal fusion were measured in 94 children: 20 with infantile ET, 16 with infantile accommodative ET, 22 with late-onset accommodative ET, 10 with intermittent infantile strabismus, and 26 normal control participants. RESULTS: Patients with infantile ET and infantile accommodative ET had high concordance between mVEP responses and stereoacuity and mVEP responses and bifoveal fusion. Asymmetric mVEP responses were highly concordant with both no measurable stereopsis and an absence of fusional vergence. Patients with late-onset accommodative ET and intermittent infantile strabismus revealed discordance between the mVEP response and stereoacuity and high concordance between the mVEP response and bifoveal fusion. Asymmetric mVEP responses were highly concordant with the absence of bifoveal fusion and the minimum-size prism to elicit fusional vergence. CONCLUSIONS: The qualitative and quantitative relationship between the mVEP response and fusional vergence suggests that the mVEP response is an objective measure of bifoveal fusion. The availability of such a test will facilitate studies of normal development of bifoveal fusion and development of monofixation syndrome in strabismus. PMID- 10670469 TI - Overexpression of MMP-1 and MMP-3 by cultured conjunctivochalasis fibroblasts. AB - PURPOSE: To determine whether conjunctivochalasis, denoting redundant, loose, nonedematous inferior bulbar conjunctiva, is associated with increased expression and activity of matrix metalloproteinases (MMPS) over their tissue inhibitors (TIMPs). METHODS: Expression of transcripts and proteins of MMPs, TIMPs, and urokinase plasminogen activator (uPA) by cultured normal human conjunctival and conjunctivochalasis fibroblasts was determined by Northern hybridization, enzyme linked immunosorbent assay (ELISA), and Western blot analysis, respectively. Gelatin and casein zymography and quantitative collagenase activity assay were performed in the serum-free conditioned media. RESULTS: Compared with normal conjunctival fibroblasts from six subjects, conjunctivochalasis fibroblasts from eight patients showed markedly increased transcript expression of MMP-1 (5- to 32 fold) and MMP-3 (4 to 30-fold), whereas that of MMP-2, TIMP-1, TIMP-2, and uPA was similar between the two groups. Protein levels were increased in the serum free conditioned media of conjunctivochalasis fibroblasts for MMP-1 (3.5- to 7.6 fold) and MMP-3 (2.3- to 13-fold), determined by ELISA and Western blot analysis. There was increased caseinolytic activity of MMP-3 and collagenolytic activity of MMP-1 (2.2-fold) by conjunctivochalasis fibroblasts, whereas no difference was noted between these two types of fibroblasts in the protein and gelatinolytic activity of MMP-2 or expression of TIMP-1 and TIMP-2 proteins, although that of TIMP-1 transcript was slightly higher in some conjunctivochalasis fibroblasts. No expression of MMP-9 was detected. CONCLUSIONS: Overexpression of MMP-1 and MMP-3 mRNA by conjunctivochalasis fibroblasts is correlated with their increased protein levels and proteolytic activities. Collectively, these data help explain how conjunctivochalasis manifests excessive degradation of the conjunctival matrix and Tenon's capsule. PMID- 10670471 TI - Response variability in the visual field: comparison of optic neuritis, glaucoma, ocular hypertension, and normal eyes. AB - PURPOSE: To compare the relationship between sensitivity and response variability in the visual field of normal eyes and eyes with optic neuritis (ON), glaucoma (POAG), and ocular hypertension (OHT). METHODS: Frequency-of-seeing (FOS) data were collected from four visual field locations in one eye of 71 subjects (12 ON, 25 POAG, 11 OHT, and 23 normal), using a constant stimulus method on an Henson 4000 perimeter (Tinsley Instruments, Croydon, UK). At each location, at least 20 stimuli (subtending 0.5 degrees) were presented for 200 ms at six or more intensities above and below the estimated threshold. The mean and SD of the probit fitted cumulative Normal function were used to estimate sensitivity and response variability. Cluster regression analysis was carried out to determine whether there were differences in the sensitivity-log (variability) relationship between the four groups. RESULTS: Variability was found to increase with decreased sensitivity for all four groups. The combined data from the four groups was well represented (R2 = 0.57) by the function log(e)(SD) = A.sensitivity (dB) + B, where the constants A and B were -0.081 (SE, +/-0.005) and 3.27 (SE, +/ 0.15), respectively. Including other statistically significant covariates (false negative errors, P = 0.004) and factors (diagnosis, P = 0.005) into the model increased the proportion of explained variance to 62% (R2 = 0.62). Stimulus eccentricity (P = 0.34), patient age (P = 0.33), fixation loss rate (P = 0.10), and false-positive rate (P = 0.66) did not reach statistical significance as additional predictors of response variability. CONCLUSIONS: The relationship between response variability and sensitivity is similar for ON, POAG, OHT, and normal eyes. These results provide supporting evidence for the hypothesis that response variability is dependent on functional ganglion cell density. PMID- 10670472 TI - Mediation of laser trabeculoplasty-induced matrix metalloproteinase expression by IL-1beta and TNFalpha. AB - PURPOSE: Laser trabeculoplasty of the anterior uveal region of the trabecular meshwork induces sustained matrix metafloproteinase expression within the juxtacanalicular region of the meshwork. Studies were conducted to test the hypothesis that a factor mediates this response and to identify the factor. METHODS: Human anterior segment organ cultures were subjected to laser treatment using standard clinical parameters and were returned to culture for 8 hours. The resultant 8-hour-conditioned culture medium was then tested for factor activity by evaluating its ability to produce two typical trabecular responses to laser treatment, that is, to induce stromelysin expression or to trigger cell division, when applied to fresh organ cultures or to cell cultures. Confocal immunohistochemistry of the laser-treated organ cultures and western immunoblot analysis of the conditioned medium were used to evaluate changes in potential candidates for the factor activity. The ability of the interleukin (IL)-1 receptor antagonist (IL-1ra)- and of tumor necrosis factor alpha (TNFalpha)- blocking antibodies to eliminate the stromelysin induction was evaluated. RESULTS: Medium conditioned for 8 hours induced typical trabecular cell division in anterior segment organ cultures. Medium conditioned for 8 hours, but not for 30 minutes, induced typical increases in stromelysin expression in these organ cultures and in cell cultures. After 8 hours, both trabecular cells in laser treated organ cultures and in the conditioned medium contained elevated levels of IL-1beta and TNFalpha. The laser-treated organ cultures contained elevated levels of IL-1alpha, but it was not secreted into the medium. The ability of conditioned media to induce stromelysin expression was partially blocked by either the IL-1ra or the TNFalpha-blocking antibody. CONCLUSIONS: Laser trabeculoplasty induces the expression and secretion of both IL-1beta and TNFalpha within the first 8 hours after treatment. These cytokines then mediate increased trabecular stromelysin expression. Putatively, this initiates remodeling of the juxtacanalicular extracellular matrix, a likely site for the aqueous outflow resistance, and thus restores normal outflow facility. PMID- 10670473 TI - Chronology of optic nerve head and retinal responses to elevated intraocular pressure. AB - PURPOSE: To determine the chronology of optic nerve head and retinal responses to elevated intraocular pressure (IOP). METHODS: After 1 to 39 days of unilaterally elevated IOP, experimental and fellow rat eyes were examined for morphology and immunohistochemical labeling alterations and for ganglion cell DNA fragmentation. RESULTS: Mean IOP for the experimental eyes was 36 +/- 8 mm Hg, an approximately 15-mm Hg elevation above normal values. By 7 days of pressure elevation above 40 mm Hg, endogenous immunostaining for brain-derived neurotrophic factor and neurotrophin 4/5 was absent from the nerve head and superior retina, whereas normal labeling was present in the inferior retina and distal optic nerve of these same eyes. These changes were preceded by a loss of gap junctional connexin43 labeling and astrocytic proliferation in the nerve head and by increased retinal ganglion cell layer apoptosis in the retina. Nerve head depletion of neurotrophins coincided with evidence of axonal degeneration, loss of astrocytic glial fibrillary acidic protein staining, and spread of collagen VI vascular immunolabeling. After longer durations at these same pressures, neurotrophin labeling returned to nerve head glia and scattered retinal ganglion cells. CONCLUSIONS: Optic nerve head and retinal responses, including the depletion of endogenous neurotrophins, are readily identified in the rat eye after experimental IOP elevation. However, the apparent chronology of these responses suggests that the withdrawal of neurotrophic support was not the only determinant of retinal ganglion cell apoptosis and axonal degeneration in response to pressure. PMID- 10670474 TI - Epithelium-deficient corneal allografts display immune privilege beneath the kidney capsule. AB - PURPOSE: To determine whether corneal tissue as an allograft displays immune privilege in a nonprivileged site and, if so, whether CD95 ligand expression contributes to the privileged status. METHODS: Syngenic and allogeneic corneal tissues deprived of epithelium were transplanted beneath the kidney capsule of normal mice. Syngeneic BALB/c, allogeneic C57BL/6, and allogeneic B6Smn.C3H-gld (CD95 ligand-deficient) mice were used as donors for BALB/c recipients, and syngeneic C3H/HeJ-gld (CD95 ligand-deficient) mice were used for normal C3H/HeJ recipients. Allogeneic conjunctival segments served as positive grafting controls. Graft fate was assessed by visual inspection at 4, 7, 14, and 21 days and was confirmed by histologic study. Viability of graft endothelium was assessed by immunocytochemical analysis. RESULTS: Syngeneic corneas and C57BL/6 corneas survived almost indefinitely beneath the kidney capsule. Both the stroma and the endothelial layers remained healthy and intact. Allogeneic conjunctiva evoked a strong inflammatory response attended by neovascularization. Similarly, B6-gld corneas deficient in CD95 ligand expression showed acute destruction beneath the kidney capsule. Circumstantial evidence implicates alloimmune rejection as the mechanism. CONCLUSIONS: Epithelium-deprived corneas from normal mice possess inherent immune privilege that protects them from alloimmune rejection even at nonprivileged sites. Constitutive expression of CD95 ligand contributes to the privileged status. It is inferred that the extraordinary success of orthotopic corneal allografts owes as much to the qualities of the graft as an immune-privileged tissue as to the qualities of the eye as an immune privileged site. PMID- 10670475 TI - Bystander activation of CD4+ T cells accounts for herpetic ocular lesions. AB - PURPOSE: Stromal keratitis is an immunopathologic consequence of herpes simplex virus (HSV) infection of the cornea. The lesion is immunopathologic, but the identities of molecules that drive the reaction remain unresolved. To exclude viral antigen recognition as a necessary step in the disease process, ocular HSV infection was followed in Tg-RAG mice (OVA-TCR transgenic mice crossed to RAG2 deficient mice) whose limited T-cell repertoire did not include immune responsiveness to HSV. METHODS: Mice with T-cell specificity to OVA peptide (Tg RAG mice) as well as control DO11.10 and BALB/c mice were infected with HSV on the scarified cornea and subjected to clinical, histologic, and immunologic analysis. To evaluate involvement of OVA-specific CD4+ T cells in lesion development in Tg-RAG mice, monoclonal antibody to CD4+ T cells was used for in vivo CD4+ T-cell depletion. RESULTS: Tg-RAG mice were capable of eliciting ocular lesions in the absence of detectable reactivity to viral antigens. Lesion manifestation in Tg-RAG mice was CD4+ T-cell dependent and the cellular infiltrates and their inflammatory products in the HSV-infected cornea were comparable to similarly infected BALB/c and DO11.10 mice. CONCLUSIONS: The authors conclude that mechanisms other than viral antigen recognition, and hence molecular mimicry, are at play and are sufficient to cause HSV-induced stromal keratitis. The data imply a significant role for non-virus-specific CD4+ T cells that could become activated by an inflammatory milieu consisting of enhanced accessory molecules and proinflammatory cytokines in the cornea. PMID- 10670476 TI - Efficacy of topical cidofovir on multiple adenoviral serotypes in the New Zealand rabbit ocular model. AB - PURPOSE: The goal of the present study was to determine the efficacy of topical 0.5% cidofovir twice daily for 7 days on the replication of multiple adenovirus (Ad) serotypes of subgroup C (Ad1, Ad5, Ad6) in the New Zealand rabbit ocular model. METHODS: In duplicate experiments for each serotype, a total of 20 rabbits (Ad5) or 16 rabbits each (Ad1 and Ad6) were inoculated topically in both eyes, with 1.5 X 10(6) pfu/eye of the appropriate virus. Twenty-four hours later, the rabbits in each serotype group were randomly divided into two topical treatment groups: I, 0.5% cidofovir; II, control vehicle. Treatment was twice daily for 7 days. All eyes were cultured for virus on days 0, 1, 3, 4, 5, 7, 9, 11, and 14. RESULTS: Compared to the control, treatment with 0.5% cidofovir reduced the following: mean Ad titer (days 1 to 7) for Ad1 (6.3 +/- 20 x 10(1) versus 2.5 +/- 3.9 X 102 pfu/ml; P < 0.0003), Ad5 (3.4 +/-5.8 x 102 versus 1.6 +/- 2.0 x 10(3) pfu/ml; P < 0.000001), and Ad6 (1.2 +/- 5.1 x 10(2) versus 5.5 +/-14 x 10(2) pfu/ml; P = 0.015); reduced Ad-positive eyes/total for Adl [45/128 (35%) versus 84/128 (66%); P = 0.000002], Ad5 [84/160 (53%) versus 131/152 (86%); P < 0.000001], and Ad6 [36/128 (28%) versus 82/128 (64%); P < 0.000001]: and reduced the duration of Ad shedding forAdl (4.9 +/-1.9 versus 9.3 +/- 3.3 days; P < 0.00007), Ad5 (6.4 +/- 2.8 versus 11.5 +/- 2.3 days; P < 0.0001), and Ad6 (4.4 +/ 2.1 versus 8.4 +/- 2.5 days; P < 0.00004). CONCLUSIONS: Topical 0.5% cidofovir twice daily for 7 days demonstrated significant antiviral activity against multiple adenoviral serotypes (Ad1, Ad5, and Ad6) in the New Zealand rabbit ocular model. These in vivo data expand in vitro studies indicating the efficacy of cidofovir against different adenovirus serotypes and support its use in clinical trials. PMID- 10670477 TI - Truncated forms of Pax-6 disrupt lens morphology in transgenic mice. AB - PURPOSE: Extensive literature shows that Pax-6 is critical for lens development and that Paxb mutations can result in aniridia in humans. In addition, it has been reported that truncated Pax-6 molecules can act as dominant-negative repressors of wild-type Pax-6 activity in cultured cells. This study was designed to determine whether Pax-6 molecules without either the activation domain (AD) or the homeodomain (HD) and the AD can function as dominant-negative repressors in vivo and alter the phenotype of the lens. METHODS: Transgenic mice were created harboring the alphaA-crystallin promoter linked to a cDNA encoding either a truncated Pax-6 without the C terminus (paired domain [PD] + homeodomain) or Pax 6 consisting of only the PD. The phenotype of the resultant animals was investigated by light and electron microscopy as well as atomic absorption spectroscopy. RESULTS: Two lines of PD + HD mice and three lines of PD mice were generated, all of which exhibit posterior nuclear and/or cortical cataracts of variable severity. The lenses from mice transgenic for either Pax-6 truncation are smaller and more hydrated than normal. Morphologically, the mice expressing the PD + HD of Pax-6 have swollen lens fibers with attenuated ball-and-socket junctions. In contrast, the lenses from mice overexpressing the PD of Pax-6 have posterior nuclear cataracts composed of cell debris, whereas the remaining fiber cells appear generally normal. CONCLUSIONS: The presence of truncated Pax-6 protein in the lens is sufficient to induce cataract in a wild-type genetic background. The simplest explanation for this phenomenon is a dominant-negative effect; however, a number of other possible mechanisms are presented. PMID- 10670478 TI - Noncontact specular microscopy of human lens epithelium. AB - PURPOSE: To obtain in vivo specular images of human lens epithelial cells (LECs) from persons with or without age-related cataract (ARC); to identify features that describe individual aspects of these complex images; to develop feature scales to quantify the severity of each feature; and to study the association of these features with LEC count, age, Lens Opacity Classification System III (LOCS III) classifications and microscopic features of lens epithelium in ARC. METHODS: One hundred fifty-two individuals underwent ophthalmic examinations and LOCS III cataract classifications. Specular images of lenses were captured using a modified noncontact corneal specular microscope (SML-2; Konan, Hyogo, Japan). Enhanced images were graded in a masked fashion, and the presence or absence and severity of each of four features in the specular image ("columnar organization," "linear furrows," "puffy clouds," and "black holes") was graded on a four-step scale. The generalized linear model with intraclass correlation was used to ascertain the statistical significance of associations between age, sex, LOCS III grade, cell count, and feature grade. Capsulorrhexis specimens from 29 patients were studied with correlative light and electron microscopy. RESULTS: LEC density declined with age and was inversely correlated with the scalar grade for puffy clouds and for the size and number of black holes. The scalar grade for columnar organization was inversely associated with the severity of posterior subcapsular and nuclear cataracts, which was the only feature associated with the LOCS III grade of ARC. No statistically significant associations were found between average cell count and LOCS III grade. CONCLUSIONS: With the use of the corneal specular microscope excellent in vivo specular images of the LECs were obtained, the features in these images that correlated well with microscopic findings were classified, and cell density in vivo was estimated. PMID- 10670479 TI - Use of a lipophilic cation to monitor electrical membrane potential in the intact rat lens. AB - PURPOSE: Tetraphenylphosphonium (TPP+) is a permeant lipophilic cation that accumulates in cultured cells and tissues as a function of the electrical membrane potential across the plasma membrane. This study was undertaken to determine whether TPP+ can be used for assessing membrane potential in intact lenses in organ culture. METHODS: Rat lenses were cultured in media containing 10 microM TPP+ and a tracer level of 3H-TPP+ for various times. 3H-TPP+ levels in whole lenses or dissected portions of lenses were determined by liquid scintillation counting. Ionophores, transport inhibitors, and neurotransmitters were also added to investigate their effects on TPP+ uptake. RESULTS. Incubation of lenses in low-K+ balanced salt solution and TC-199 medium, containing physiological concentrations of Na+ and K+, led to a biphasic accumulation of TPP+ in the lens that approached equilibrium by 12 to 16 hours of culture. The TPP+ equilibrated within 1 hour in the epithelium but penetrated more slowly into the fiber mass. The steady state level of TPP+ accumulation in the lens was depressed by 90% when the lenses were cultured in a medium containing high K+. The calculated membrane potential for the normal rat lens in TC-199 was -75 +/- 3 mV. Monensin (1 microM) and nigericin (1 microM), Na+H+ and K+H+ exchangers respectively, as well as the protonophore carbonylcyanide-m-chlorophenylhydrazone (CCCP, 10 microM) and the calcium ionophore A23187 (10 microM), abolished TPP+ accumulation and caused cloudiness of the lenses. The neurotransmitter acetylcholine at 50 microM decreased TPP+ accumulation in the lens, but this effect could be prevented by simultaneous application of 1 mM atropine. CONCLUSIONS: TPP+ accumulation can be used as an indicator of changes in membrane potential in intact lenses, but because of the long time required to reach steady state, its utility is limited. The slow accumulation of TPP+ and its slow efflux from the lens under conditions known to depolarize membranes are consistent with a diffusion barrier in the deep cortex and nucleus of the lens. PMID- 10670481 TI - Autofluorescence distribution associated with drusen in age-related macular degeneration. AB - PURPOSE: To determine whether drusen in patients with age-related maculopathy and macular degeneration (ARM/AMD) are associated with focal changes in retinal pigment epithelium (RPE) lipofuscin fluorescence. METHOD: A new autofluorescence imaging device was used to study lipofuscin distribution associated with individual drusen in 20 patients with ARM/AMD. Paired monochromatic and autofluorescence fundus images were used for detailed analysis of the topography of autofluorescence at specific sites containing drusen. In four eyes, image analysis was used to compare the spatial distribution of the autofluorescence with the location of drusen and to quantify the autofluorescence distribution over individual drusen (54 drusen). REsuLTs. A specific pattern of autofluorescence was frequently found to be spatially associated with hard drusen and soft drusen between 60 and 175 microm in size. The pattern is characterized by a central area of decreased autofluorescence surrounded, in most cases, by an annulus of increased autofluorescence. The location of this pattern was highly correlated with the position of individual distinct drusen. The central low autofluorescence focus was on average 16% below the surrounding background, and the annulus, when present, was on average 6% more fluorescent than the background. Soft drusen larger than 175 microm and confluent soft drusen show either multifocal areas of low autofluorescence or a more heterogeneous distribution. CONCLUSIoNs. Autofluorescence imaging permits measurement of RPE lipofuscin at specific sites. RPE overlying drusen have altered autofluorescence, suggesting changes in RPE health. PMID- 10670480 TI - Basolateral Na(+)-K(+)-2Cl(-) cotransport in cultured and fresh bovine corneal endothelium. AB - PURPOSE: To examine whether Na(+)-K(+)-2Cl(-) cotransport has the potential to contribute to corneal endothelial ion and fluid transport in cultured and fresh bovine corneal endothelial cells. METHODS: Cl- and Na+ sensitive fluorescent dyes were used to measure furosemide-dependent ion fluxes in cultured and fresh endothelial cells. Immunoblot analysis and immunofluorescence were used to determine expression and location of the Na(+)-K(+)-2Cl(-)cotransporter (NKCC1). RESULTS: Application of furosemide (50-100 microM) reduced Cl- and Na+ influx in approximately 50% of trials using cultured cells and only 10% of trials with fresh cells; however, in all cases pretreatment with furosemide slowed Cl- efflux when cells were bathed in Cl(-)-free Ringer's. Double-sided perfusion of cultured cells indicated that furosemide-sensitive Cl- fluxes were located on the basolateral side. Immunoblot analysis revealed 174-kDa bands in both fresh and cultured cells, but the bands were denser in fresh endothelial cells. Immunofluorescence showed distinct lateral membrane staining in addition to significant amounts of perinuclear staining. CONCLUSIONS: The Na(+)-K(+)-2Cl(-) cotransporter is present in both fresh and cultured bovine corneal endothelium, and the expression is apparently higher in the fresh cells. The cotransporter is present on the lateral membrane consistent with a role in loading endothelial cells with Cl-, thereby possibly contributing to a transendothelial Cl- flux. However, in the resting cell, net flux through the transporter is often not apparent. PMID- 10670482 TI - Erbium: YAG laser ablation of retinal tissue under perfluorodecaline: determination of laser-tissue interaction in pig eyes. AB - PURPOSE: To evaluate the effect of Er;YAG laser on pig retina using a perfluorodecaline/retina interphase with the goal of precisely determining the extent of retinal tissue ablation. METHODS: Free running (tau = 250 microsec) Er:YAG laser pulses were transmitted through a zirconium fluoride (ZrF4) fiber guarded by quartz rod (d = 1000 microm). Laser pulses were applied to the retinal surface of enucleated pig eyes. Eyes were mounted in a specially designed rotating sample holder. The fiber probe was elevated 1.0 +/- 0.3 mm above the retinal surface with perfluorodecaline serving as transmitting medium. The laser energy was applied in a circular pattern with a radius of 3.0 mm. Radiant exposures were set to 1, 3, 5, and 10 J/cm2. RESULTS: Tissue ablation linearly increased with radiant exposure from 3.2 +/- 3.7 microm at 1 J/cm2 up to 40.9 +/- 12.9 microm at 10 J/cm2. Thermal tissue changes extended 70 +/- 10 microm vertically into the retina and 25 +/- 5 microm horizontally. Distortion of outer photoreceptor segments was noticed when the retina was exposed to radiant exposures of 3 J/cm2 or higher. CONCLUSIONS: The Er:YAG laser in combination with perfluorodecaline produced precise ablation of the pig retina, which suggests the feasibility of this technique for safe ablation of epiretinal membranes. PMID- 10670483 TI - Multifocal electroretinogram in occult macular dystrophy. AB - PURPOSE: Occult macular dystrophy (OMD) is an unusual macular dystrophy presenting with an essentially normal fundus and fluorescein angiography but with progressive central visual loss. The authors studied the function of local retinal areas in the posterior pole of patients with OMD using multifocal electroretinograms (ERGs). METHODS: Multifocal ERGs were recorded using the Visual Evoked Response Imaging System with 61 hexagonal elements within a visual field of 30 degrees radius from 8 OMD patients and 20 age-matched, normal subjects. The amplitudes and implicit times of the patients and normal control subjects were compared at the various retinal eccentricities. RESULTs. The amplitudes of the multifocal ERGs in the OMD patients were markedly reduced in the central 7 degrees of the fovea. The difference of the ERG amplitudes between OMD and normal subjects became smaller toward the peripheral retina. Most OMD patients had slight but significantly delayed implicit times across the whole testing field, and the differences between the OMD and the normal subjects did not change with retinal eccentricity. CONCLUSIONS: Our results for multifocal ERG amplitudes support the idea that OMD patients have localized retinal dysfunction distal to the ganglion cells in the central retina. The delayed implicit times across the whole test field suggest that the retinal dysfunction has a broader boundary than expected by ERG amplitudes and psychophysical perimetric results. PMID- 10670484 TI - Schwann cell grafting into the retina of the dystrophic RCS rat limits functional deterioration. Royal College of Surgeons. AB - PURPOSE: To examine whether congenic Schwann cells grafted into the subretinal space of dystrophic Royal College of Surgeons (RCS) rats can prevent photoreceptor loss and maintain visual function. METHODS: Purified neonatal Schwann cells derived from congenic rats were grafted into the subretinal space of 3- to 4-week-old dystrophic RCS rats. Graft placement was confirmed using Schwann cells labeled in vitro with the fluorescent dye Hoechst 33342 or in grafted eyes processed for electron microscopy (48-hour to 1-month survival). At longer intervals, up to 9 months after surgery, animals were examined for photoreceptor survival; preservation of a visual reflex, head-tracking to moving stripes; and preservation of visual receptive fields associated with the region of graft placement. RESULTS: One week after the graft was performed, Schwann cells had integrated into the subretinal space with little evidence of a reactive response. When screened for head-tracking to moving stripes, Schwann cell-grafted animals performed better than sham-treated or control dystrophic animals. Threshold sensitivity measurements and visual field assessment made by recording from the superior colliculus also showed a significant level of preserved function compared with control animals. Functional rescue was correlated with photoreceptor survival and could be observed for at least 9 months after grafting. CONCLUSIONS: Schwann cells injected into the subretinal space limit functional deterioration and prolong photoreceptor survival. It is suggested that they act by local release of growth factors that either support photoreceptors directly and/or stimulate phagocytosis in RPE cells. PMID- 10670485 TI - Tractional force generation by porcine Muller cells: paracrine stimulation by retinal pigment epithelium. AB - PURPOSE: To examine the ability of retinal pigment epithelial (RPE) cells to modulate Muller cell extracellular matrix contraction through secreted promoters. METHODS: Freshly isolated RPE cells were maintained in continuous culture until the morphologic and immunocytochemical changes associated with myofibroblastic dedifferentiation were complete. Secretory products collected from these cells during extended incubations in serum-free medium and at different stages of dedifferentiation were examined for the ability to promote extracellular matrix contraction by Muller cells. The contributions of specific growth factors to RPE secreted activity were examined with growth factor-neutralizing antibodies. RESULTS: Secretory products from RPE cells throughout dedifferentiation contained biologically active quantities of Muller cell contraction promoters. Secretory activity increased during extended incubation in serum-free medium and during myofibroblastic dedifferentiation. Growth factor-specific neutralizing antibodies enabled the determination that insulin-like growth factor- and platelet-derived growth factor-related proteins were the secreted species to which Muller cells responded. Finally, gene expression of insulin-like growth factor 1 and platelet derived growth factor A chain by porcine RPE cells was confirmed using reverse transcription-polymerase chain reaction. CONCLUSIONS. RPE cells are a viable source of biologically active quantities of two growth factors that stimulate extracellular matrix contraction by Muller cells. This secretory profile persists for extended periods in an otherwise serum-free environment and is enhanced during myofibroblastic dedifferentiation. PMID- 10670486 TI - Progressive optic axon dystrophy and vacuslar changes in rd mice. AB - PURPOSE: To examine how the vascular plexuses in the rd mouse retina are affected by the loss of photoreceptors and how this compares with the Royal College of Surgeons (RCS) rat. To examine whether the profound effects of vascular pathology on retinal ganglion cells (RGCs) and their axons seen in RCS rats are also found in rd mice. METHODS: Vascular patterns were studied in flatmounted and sectioned retinas using either nicotinamide adenine dinucleotide phosphate(NADPH) diaphorase histochemistry or vessel filling with horseradish peroxidase. Optic axons were visualized using RT97 (an antibody against the 200-kDa neurofilament subunit), and RGCs were labeled by retrograde transport of fluorescence label, the Fluorogold, applied to the superior colliculus. RESULTS: The present study showed that in the rd mouse, similar to the RCS rat, vascular complexes developed in association with retinal pigment epithelial cells at the outer border of the retina. The number and distribution of complexes were very different from the rat, but as in the rat, progressive axonal dystrophy was seen in the optic fiber layer. RGC loss, rather than being local was more broadly distributed, but some, at least, appeared to be secondary to axonal dystrophy caused by vessels supplying vascular formation. CONCLUSIONS: Photoreceptor loss in the rd mouse leads to RGC axonal dystrophy and loss. The lesser degree and different distribution of RGC loss caused by abnormal vasculature associated with vascular formations in the outer retina in the rd mouse may be due to the early atrophy of the deep vascular plexus in this animal. PMID- 10670487 TI - Differential expression of cadherin adhesion receptors in neural retina of the postnatal mouse. AB - PURPOSE: To determine the expression pattern of multiple subtypes of cadherin adhesion receptor in postnatal mouse neural retina. METHODS: The expression of N cadherin, R-cadherin, cadherin-6, cadherin-8, and cadherin-11 in retinas at postnatal days 0 to 42 was analyzed by in situ hybridization of mRNA as well as by immunohistochemistry. RESULTS: Each cadherin was expressed by different cell populations of the retina, and the following expression patterns were established by postnatal day 14: in the ganglion cell layer, all these molecules were expressed, but each occurred only in a subset of the cells. Likewise, in the inner nuclear layer, R-cadherin and cadherin-6 and -8 were expressed by a restricted population of amacrine cells, and cadherin-8 also by a subpopulation of bipolar cells. All horizontal cells expressed R-cadherin, and Muller cells expressed N-cadherin and cadherin-11. Proteins of R-cadherin and cadherin-6 were concentrated in neuropil layers. CONCLUSIONS: The pattern of differential expression of the five cadherins supports the idea that these molecules may play a role in selective cell interactions within the heterogeneous cell pool of the neural retina. PMID- 10670488 TI - Expression of ciliary neurotrophic factor activated by retinal Muller cells in eyes with NMDA- and kainic acid-induced neuronal death. AB - PURPOSE: To elucidate the role of retinal Muller cells in N-methyl-D-aspartate (NMDA)- or kainic acid (KA)induced retinal damage. METHODS: In experimental eyes, NMDA or KA was injected into the vitreous of rat eyes. Immunohistochemistry and western blot analysis were conducted to elucidate expression and localization of glial fibrillary acidic protein (GFAP) and ciliary neurotrophic factor (CNTF). In addition, the neuroprotective effects of CNTF were calculated by counting cells in the ganglion cell layer (GCL) and by measuring the thickness of the various retinal layers. RESULTS: Morphometric analysis of retinal damage in NMDA- and KA injected eyes showed significant cell loss in the GCL and thinning of the inner plexiform layer (IPL) of the retina, but not of other retinal layers. Immunohistochemistry demonstrated disappearance and/or decrease in immunoreactivities of calbindin- and calretinin- positive cells and their neurites and upregulated expression of both GFAP and CNTF in experimental eyes. Western blot analysis showed an increase in protein expression for CNTF in retinas of experimental eyes. Confocal images and sequential localization demonstrated colocalization of CNTF and GFAP in the inner retinal layer and possibly in Muller cells. In addition, pretreatment with CNTF (1 microg) before the intravitreal injection of NMDA (or KA) demonstrated that CNTF has neuroprotective effects against NMDA- or KA-induced neuronal death in the retina. CONCLUSIONS: These studies revealed the upregulated expression of CNTF and GFAP in Muller cells in response to NMDA- and KA-induced neuronal death, suggesting that production of CNTF in Muller cells may be a part of the endogenous neuroprotective system in the retina. PMID- 10670489 TI - Production and accumulation of thrombospondin-1 in human retinal pigment epithelial cells. AB - PURPOSE: To investigate the production and release of thrombospondin-1 (TSP-1), a natural inhibitor of angiogenesis, by human retinal pigment epithelial (RPE) cells to clarify the possible role of TSP-1 in maintaining intraocular angiogenesis. METHODS: Human RPE cells were isolated from a human cadaveric eye and cultured in medium with 5% newborn calf serum. TSP-1 messages in the purified RNA of RPE cells were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Intracellular TSP-1 peptides were detected by cytofluorographic analysis. TSP-1 peptides in the culture medium on RPE cells were measured by sandwich enzyme-linked immunosorbent assay (ELISA). TSP-1 specific immunofluorescent staining was tested in RPE cells cultured on glass slides and in a human retinal tissue specimen. RESULTS: mRNA specific for TSP-1 was found in RT-PCR products from RPE cells, and it showed a time-dependent increase from the beginning of the culture. Intracellular staining for TSP-1 was identified by flow cytometry. The sandwich ELISA identified a time-dependent increase of TSP-1 peptides in the culture medium of RPE cells. Immunostaining for TSP-1 was observed in the cytoplasm of RPE cells cultured on glass slides. Positive immunostaining of TSP-1 was observed in the cytoplasm of the RPE layer in the human retinal tissue specimen. CONCLUSIONS: RPE cells can produce and release TSP 1 in vitro, and TSP-1 accumulates in the cytoplasm of RPE cells in vivo as well as in vitro. The production of TSP-1 by RPE cells is influenced by the state of proliferation and/or cell density. TSP-1 appears to be an important control factor in retinal and choroidal neovascularization. PMID- 10670490 TI - Rapid upregulation of fibroblast growth factor receptor 1 (flg) by rat photoreceptor cells after injury. AB - PURPOSE: To determine the mechanism by which basic fibroblast growth factor (bFGF) exerts its neuroprotective effects on degenerating or injured photoreceptors. METHODS: Confocal immunofluorescence microscopy was used to identify sites of bFGF and FGF receptor 1 (FGFR1) expression after focal injury or experimental retinal detachment in adult rats. FGFR1 expression was analyzed immunohistochemically and at the transcription level in single photoreceptor cells, after reverse transcription (RT), using the polymerase chain reaction (PCR). Real time quantitative RT-PCR was used to measure changes in FGFR1 mRNA levels in the retina in response to injury or detachment. RESULTS: Confocal immunofluorescence observations showed that FGFR1 immunoreactivity in the rat retina is concentrated primarily in the perinuclear cytoplasm of photoreceptor cell bodies. Reverse transcription of total RNA derived from dissociated rat photoreceptor cells, followed by amplification of FGFR1 cDNA using the PCR, verified the presence of FGFR1 transcripts in normal rat photoreceptor cells; in contrast, no evidence of bFGF transcription was detected. Collectively, these results provide compelling evidence for FGFR1 gene expression by rat photoreceptors in situ. Within hours after experimental retinal detachment or focal injury, there is a twofold increase in FGFR1 immunoreactivity in the outer nuclear layer that persists for at least 7 days; a similar increase in bFGF immunoreactivity in the interphotoreceptor matrix is also apparent. This increase in FGFR1 protein levels after detachment and injury also was confirmed by western blot analysis. Real time quantitative RT-PCR analyses revealed that a rapid upregulation of FGFR1 mRNA occurred within 12 hours after retinal injury/detachment, but then declined to near baseline levels by 24 hours. CONCLUSIONS: This body of evidence strongly suggests that the photoreceptor rescue effect elicited by retinal injury as well as by injection of exogenous bFGF is mediated, at least in part, by upregulation of the FGFR1 by the photoreceptor cells. PMID- 10670491 TI - The use of adenovirus-mediated gene transfer to develop a rat model for photoreceptor degeneration. AB - PURPOSE: To investigate the effects of recombinant adenovirus-mediated downregulation of cathepsin S (CatS) on the retinal pigment epithelium and/or neural retina in vivo. METHODS: The expression of green fluorescent protein (gfp) after subretinal injection of a recombinant adenovirus, Ad.gfp, into rat eyes was first established by in vivo fundus fluorescence photography and fluorescence microscopy. The autofluorescent debris accumulation in Ad.CatSAS (recombinant adenovirus carrying the antisense CatS gene)injected rat eyes was monitored by fluorescence microscopy, and the antisense CatS RNA expression was demonstrated by in situ hybridization. Changes in the retinal morphology were assessed by light microscopy. ResuLTS. The gfp expression was present in 30% to 90% of the injection area at 3 days and was absent 9 days after Ad.gfp injection. In Ad.CatSAS-injected eyes, the expression of antisense CatS RNA was demonstrated by in situ hybridization. Autofluorescent debris accumulation was significantly higher in Ad.CatSAS-injected eyes than in control eyes. The shortening of photoreceptor outer segments in Ad.CatSAS-injected eyes coincided with intense autofluorescent debris accumulation. The number of layers of photoreceptor cells decreased with time and were 11, 9, and 8 at 7, 14, and 28 days after Ad.CatSAS injection, respectively. In control eyes, the number of layers of photoreceptor cells (14) remained unchanged. CONCLUSIONS: These results demonstrate that recombinant adenovirus-mediated transient modulation of gene expression in retinal pigment epithelial (RPE) cells could induce changes in the retina, and, in spite of the low expression of endogenous CatS in RPE cells, this enzyme plays an important role in maintenance of normal retinal function. PMID- 10670492 TI - Antisense knockdown of GLAST, a glial glutamate transporter, compromises retinal function. AB - PURPOSE: To elucidate the role of the glial glutamate transporter GLAST, in the regulation of retinal function. METHODS: Antisense oligonucleotides to GLAST were injected intravitreally into the left eye of Wistar rats. Sense oligonucleotides (control) were injected into the right eye over a period of 3 days. Scotopic flash electroretinograms were recorded over a 20-day period. To assay whether the antisense oligonucleotides caused a reduction in the expression or the activity of GLAST, retinas were exposed to D-aspartate, a nonendogenous substrate of glutamate transporters. The retinas were immunolabeled with specific antibodies for D-aspartate. Retinal GLAST and glutamate distributions also were determined immunocytochemically. RESULTS: Antisense oligonucleotides markedly suppressed the electroretinogram b-wave, whereas sense oligonucleotides had no significant effect. Significant changes in the electroretinogram were apparent 5 days after injection of antisense oligonucleotide and were sustained for at least 20 days. A marked reduction of D-aspartate uptake into Muller cells of retinas that had been exposed to the antisense oligonucleotides 5 days previously suggests a reduction of GLAST activity. The retinas, however, displayed no evidence of excitotoxic neuronal degeneration, and the distribution of glutamate was unaffected by antisense treatment. CONCLUSIONS: The observed lack of neuronal degeneration suggests that reduced glutamate uptake into Muller cells does not cause excitotoxic tissue damage. A direct perturbation of glutamatergic signaling is more likely, because the rapid clearance of glutamate is necessary for light elicited signaling between photoreceptors and bipolar cells. This suggests that GLAST is essential for the maintenance of normal retinal transmission. PMID- 10670493 TI - Bone morphogenetic proteins-2 and -4: negative growth regulators in adult retinal pigmented epithelium. AB - PURPOSE: To determine the relative level and localization of bone morphogenetic protein (BMP-4 mRNA in the retina and retinal pigmented epithelium (RPE) under normal and pathologic conditions, to seek clues regarding possible functions. METHODS: Clones isolated from an RPE cDNA library were sequenced and used as probes for northern blot analysis. Expression in the retina and RPE was investigated in mouse models using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. The effect of recombinant proteins on RPE proliferation was investigated by thymidine incorporation. RESULTS: Bovine clones with high homology to BMP-2 and BMP4 were isolated from a subtracted RPE cDNA library. Northern blot analysis using the clones as probes demonstrated abundant and differential expression in adult bovine RPE, but with RT-PCR and in situ hybridization, expression was also demonstrated in mouse retinal neurons. In mice with oxygen-induced ischemic retinopathy there was a striking decrease in BMP-4 mRNA in the retina within 6 hours of the onset of hypoxia that was maintained for at least 5 days. In mice with inherited photoreceptor degeneration, there was a dramatic decrease in BMP4 mRNA in retina and RPE during and after the degeneration. mRNA for the type II BMP receptor was observed in freshly isolated and cultured RPE cells, isolated retina, and freshly isolated bovine aortic endothelial cells. Thymidine incorporation in early-passage RPE cells showed a 14-fold stimulation above control with 5% serum that was decreased to 322%, 393%, and 313% in the presence of BMP-2 (10 ng/ml), BMP4 (10 ng/ml), and transforming growth factor (TGF)-,1 (2 ng/ml), respectively. CONCLUSIONS: BMP-2 and BMP-4 may serve as negative growth regulators in the retina and RPE that are downregulated by injury, to allow tissue repair. Modulation of expression of the BMPs may provide a means to control the exaggerated wound repair that occurs in proliferative retinopathies. PMID- 10670494 TI - Human platelet suspension stimulates porcine retinal glial proliferation and migration in vitro. AB - PURPOSE: To characterize the cellular and molecular mechanisms underlying the efficacy of autologous platelet suspension adjuvant therapy in the treatment of macular hole. METHODS: Platelet suspensions were: paid from whole blood samples obtained from informed volunteers. For proliferation assays, platelet suspensions or purified growth factors were added to semi-confluent cultures of porcine real glial cells for 24 hours, followed by [3H]thymidine for 15 hours, after which time cells were washed, solubilized, and counted for uptake of radioactive tracer. For cell migration assays, confluent glial cultures were scrape wounded and maintained in the presence or absence of platelet suspension or identified platelet constituents. Cell migration into the denuded area was scored as a function of time. In certain cases, specific pharmacologic inhibitors of growth factor action were added at the same time as platelet adjuvant or growth factors. RESULTS: Platelet suspension adjuvant induced strong mitogenic and chemotactic responses in cultured glia, in a dose-dependent manner. Maximal incorporation of thymidine was two- to threefold that of control levels, with an ED50 approximately 5 x 10(6) platelets/ml, and migration was enhanced up to 80-fold after 48 hours. Platelet suspension-induced proliferation was completely blocked by addition of 25 microM genistein, a tyrosine kinase receptor inhibitor. However, the same concentration only partially blocked the cell migration response. Addition of any single growth factor or protein identified from ELISA analysis, or a combination of all factors, did not significantly stimulate proliferation or cell migration. CONCLUSIONS: Human platelet suspensions exert both proliferative and chemotactic influences on retinal glial cells in vitro, suggesting that the same responses may occur in platelet-induced macular hole repair in humans. Growth factors or proteins that have been identified within the suspensions do not mimic these responses in vitro, implying that additional currently unidentified trophic activities are also present. PMID- 10670495 TI - Noninvasive assessment of retinal function in rats using multifocal electroretinography. AB - PURPOSE: To assess the applicability of multifocal electroretinograms (mfERGs) for evaluation of function in this small-eyed animal with a rod-dominant retina that is often used to model retinal diseases. METHODS: Noninvasive monocular mfERGs were recorded in anesthetized albino (Sprague-Dawley) and pigmented (Long Evans) rats. Achromatic stimuli subtending a 49 degrees by 53 degrees field consisted of 61 hexagons that were generated and presented (at varying rates and luminances) using a Visual Evoked Response Imaging System (VERIS; EDI, San Mateo, CA). The VERIS also was used to calculate individual responses and for analysis. RESULTS: mfERGs were recorded from pigmented and albino rats by slowing the rate of stimulus presentation to allow for the slow recovery time of the rod system. In each rat strain, responses varied systematically with changes in stimulus parameters. Peak response amplitude increased as the rate of stimulation was slowed and as stimulus luminance was increased. Response latency decreased as stimulus intensity was increased. The local nature of the response was assessed by several independent measures. CONCLUSIONS: The present work demonstrated the feasibility and limitations of using mfERG to assess topographical changes in the rat retina. It showed that despite the problems of the unavoidable self-adapting nature of the stimulus, the small eye of the animal, and the high potential for light scatter within the retina, multifocal responses with a good signal-to-noise ratio can be obtained from the rat. PMID- 10670496 TI - Comments on vasectomy closure techniques. PMID- 10670498 TI - Comments on vasectomy closure techniques. PMID- 10670497 TI - Comments on vasectomy closure techniques. PMID- 10670499 TI - Who should operate in carotid disease? PMID- 10670500 TI - Controversy in otitis media management: should we follow the CDC recommendations? PMID- 10670501 TI - Acute otitis media caused by resistant pneumococci. PMID- 10670502 TI - Topical therapy for acne. AB - Acne is a common problem in adolescents and young adults. The disorder is caused by abnormal desquamation of follicular epithelium that results in obstruction of the pilosebaceous canal. This obstruction leads to the formation of comedones, which can become inflamed because of overgrowth of Propionibacterium acnes. Topical retinoids such as tretinoin or adapalene are effective in many patients with comedonal acne. Patients with inflammatory lesions benefit from treatment with benzoyl peroxide, azelaic acid or topical antibiotics. Frequently, the use of comedonal and antibacterial agents is required. PMID- 10670503 TI - Urinary catheter management. AB - The use of urinary catheters should be avoided whenever possible. Clean intermittent catheterization, when practical, is preferable to long-term catheterization. Suprapubic catheters offer some advantages, and condom catheters may be appropriate for some men. While clean handling of catheters is important, routine perineal cleaning and catheter irrigation or changing are ineffective in eliminating bacteriuria. Bacteriuria is inevitable in patients requiring long term catheterization, but only symptomatic infections should be treated. Infections are usually polymicrobial, and seriously ill patients require therapy with two antibiotics. Patients with spinal cord injuries and those using catheters for more than 10 years are at greater risk of bladder cancer and renal complications; periodic renal scans, urine cytology and cystoscopy may be indicated in these patients. PMID- 10670504 TI - Update on the prevention and treatment of sexually transmitted diseases. AB - The Centers for Disease Control and Prevention updated its guidelines for the treatment of sexually transmitted diseases. The guidelines include the following information: recommendations for hepatitis A immunization and expanded indications for hepatitis B vaccination; updated diagnostic criteria for pelvic inflammatory disease and parenteral treatment regimens; information on two additional antiviral agents for the treatment of genital herpes; a recommendation for use of a single 1-g dose of azithromycin (Zithromax) to treat urethritis and chlamydial cervicitis; information on the use of quinolones in the treatment of gonococcal infections; information on podofilox and imiquimod, which are both patient-applied medications, in the treatment of noncervical human papillomavirus infection; updated guidelines for the prevention and detection of congenital syphilis; and information on how to prevent the spread of sexually transmitted diseases by educating patients about the importance of changing their sexual behaviors. To have a significant impact on the current rate of transmission of sexually transmitted diseases, family physicians should develop a plan to integrate the guidelines into their practices. PMID- 10670505 TI - Perianal streptococcal dermatitis. AB - Perianal streptococcal dermatitis is a bright red, sharply demarcated rash that is caused by group A beta-hemolytic streptococci. Symptoms include perianal rash, itching and rectal pain; blood-streaked stools may also be seen in one third of patients. It primarily occurs in children between six months and 10 years of age and is often misdiagnosed and treated inappropriately. A rapid streptococcal test of suspicious areas can confirm the diagnosis. Routine skin culture is an alternative diagnostic aid. Treatment with amoxicillin or penicillin is effective. Follow-up is necessary, because recurrences are common. PMID- 10670506 TI - When to operate in carotid artery disease. AB - Carotid endarterectomy has proved to be beneficial in the prevention of stroke in selected patients. The procedure is indicated in symptomatic patients with carotid-territory transient ischemic attacks or minor strokes who have carotid artery stenosis of 70 to 99 percent. With a low surgical risk, carotid endarterectomy provides modest benefit in symptomatic patients with carotid artery stenosis of 50 to 69 percent. Platelet antiaggregants and risk factor modification are recommended in symptomatic patients with less than 50 percent stenosis. In the Asymptomatic Carotid Atherosclerosis Study, carotid endarterectomy was beneficial in patients who had asymptomatic carotid artery stenosis of 60 percent or greater and whose general health made them good candidates for elective surgery, provided that the arteriographic and surgical complication rates were low. However, in asymptomatic patients, surgery reduced the absolute risk of stroke by only 1 percent per year. PMID- 10670507 TI - Knee braces: current evidence and clinical recommendations for their use. AB - Methods of preventing and treating knee injuries have changed with the rapid development and refinement of knee braces. Prophylactic knee braces are designed to protect uninjured knees from valgus stresses that could damage the medial collateral ligaments. However, no conclusive evidence supports their effectiveness, and they are not recommended for regular use. Functional knee braces are intended to stabilize knees during rotational and anteroposterior forces. They offer a useful adjunct to the treatment and rehabilitation of ligamentous knee injuries. Patellofemoral knee braces have been used to treat anterior knee disorders and offer moderate subjective improvement without significant disadvantages. Additional well-designed studies are needed to demonstrate objectively the benefits of all knee braces. Knee braces should be used in conjunction with a rehabilitation program that incorporates strength training, flexibility, activity modification and technique refinement. PMID- 10670508 TI - Smell and taste disorders: a primary care approach. AB - Smell and taste disorders are common in the general population, with loss of smell occurring more frequently. Although these disorders can have a substantial impact on quality of life and may represent significant underlying disease, they are often overlooked by the medical community. Patients may have difficulty recognizing smell versus taste dysfunction and frequently confuse the concepts of "flavor" and "taste." While the most common causes of smell disturbance are nasal and sinus disease, upper respiratory infection and head trauma, frequent causes of taste disturbance include oral infections, oral appliances (e.g., dentures), dental procedures and Bell's palsy. Medications can interfere with smell and taste, and should be reviewed in all patients with reported dysfunction. In addition, advancing age has been associated with a natural impairment of smell and taste ability. A focused history and a physical examination of the nose and mouth are usually sufficient to screen for underlying pathology. Computed tomographic scanning or magnetic resonance imaging of affected areas, as well as commercially available standardized tests, may be useful in selected patients. The causes of olfactory dysfunction that are most amenable to treatment include obstructing polyps or other masses (treated by excision) and inflammation (treated with steroids). Enhancement of food flavor and appearance can improve quality of life in patients with irreversible dysfunction. PMID- 10670509 TI - 1999 USPHS/IDSA Guidelines for the Prevention of Opportunistic Infections in Persons Infected with HIV: Part II. Prevention of the first episode of disease. U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention. U.S. Public Health Service/Infectious Diseases Society of America. PMID- 10670510 TI - The U.S. blood supply. PMID- 10670511 TI - American Thoracic Society issues consensus statement on sarcoidosis. PMID- 10670512 TI - ACOG practice bulletin on management of herpes in pregnancy. American College of Obstetricians and Gynecologists. PMID- 10670513 TI - AHA examines cardiovascular problems in diabetes. American Heart Association. PMID- 10670514 TI - Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. PMID- 10670515 TI - Risky business? Helping the HIV-infected have babies. PMID- 10670516 TI - Rationale, interpretation, validation, and uses of sperm function tests. PMID- 10670517 TI - DNA integrity in human spermatozoa: relationships with semen quality. AB - The literature contains conflicting evidence regarding the existence of DNA damage in spermatozoa from infertile male patients. To examine this phenomenon, we have studied ejaculated spermatozoa from normozoospermic semen donors and from a group of the unselected male partners of couples attending an infertility clinic for initial investigation. Classical semen analysis according to World Health Organization (WHO) guidelines was undertaken with computer-assisted sperm analysis (CASA). Spermatozoa were prepared by sequential washing and centrifugation and were analyzed for DNA fragmentation using three assays: 1) a single-cell gel electrophoresis (comet) assay, 2) in situ nick translation with prior chemical decondensation (ISNT-decondensed), and 3) in situ nick translation without prior chemical decondensation (ISNT-condensed). In addition, reactive oxygen species (ROS) generation by spermatozoa was measured, and seminal plasma was analyzed for its total reactive antioxidant potential (TRAP). When the donor and patient groups were compared, the latter had lower levels of semen quality and higher levels of DNA damage, which was particularly apparent using the comet assay. Highly significant negative correlations were observed between DNA fragmentation, detected by all three assays, and semen quality, particularly sperm concentration. In addition, multiple regression analysis indicated that other attributes of semen quality, such as sperm movement and ROS generation, were also related to DNA damage. We conclude that a significant proportion of infertile men have elevated levels of DNA damage in their ejaculated spermatozoa. PMID- 10670518 TI - In vitro and xenogenous capacitation-like changes of fresh, cooled, and cryopreserved stallion sperm as assessed by a chlortetracycline stain. AB - Like the human female, the mare experiences reproductive tract pathology that may sometimes be circumvented by the use of assisted reproductive technologies (ARTs). One such technology, gamete intrafallopian transfer (GIFT), may be used in mares that exhibit ovulatory, oviductal, or uterine abnormalities that limit the use of common ARTs, such as embryo transfer. Homologous GIFT has been successfully performed in the horse; however, the logistics, costs, and associated risks of surgically transferring gametes to the oviducts of a recipient mare are considerably high. Use of a less costly species in a heterologous or xenogenous procedure would therefore be beneficial. This study represents the preliminary investigation into the use of sheep as recipients for xenogenous GIFT procedures using equine gametes. We investigated the capacitation response of fresh, cooled, or frozen stallion sperm after 1) in vivo incubation in the reproductive tract of estrous and anestrous ewes as well as 2) in vitro incubation in a modified Krebs/ Ringer extender at 37 degreesC with and without the addition of heparin at 10 IU/mL for up to 8 hours. A chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm. Findings indicated that oviductal fluid samples recovered from estrous ewes had significantly higher numbers of sperm exhibiting capacitation-like staining patterns when compared to samples recovered from anestrous ewes (P < .05). Fresh semen yielded higher capacitation-like staining patterns after in vivo incubation than did frozen-thawed or cooled samples. A transition from majority CTC unreacted sperm to majority CTC non-acrosome intact sperm was demonstrated for both in vivo and in vitro studies. In vitro incubation of stallion sperm with heparin did not result in an increased capacitation-like staining response over time when compared with nonheparinized samples. Results from this study suggest that xenogenous capacitation of stallion sperm may occur in the estrous ewe. PMID- 10670519 TI - Comparison of zinc concentrations in blood and seminal plasma and the various sperm parameters between fertile and infertile men. AB - The aim of the study was to examine the relationships between concentrations of zinc in blood and seminal plasma and sperm quality among infertile and fertile men. One hundred seven male (infertile group) partners of couples who were undergoing investigation for infertility with no known cause for the infertility and 103 men (fertile group) whose wives were pregnant at the time of the study were recruited. The subjects' blood and seminal plasma concentration of zinc were determined by atomic absorption spectroscopy. Except for semen volume, all the other semen parameters for the infertile men were significantly lower than those for the fertile group. The geometric means of the seminal plasma zinc concentration were significantly lower in the infertile group compared with those in the fertile group; 183.6 mg/L (range, 63-499) versus 274.6 mg/L (range, 55 420). There were no significant differences in the geometric means of the blood zinc concentration between the 2 groups. Seminal plasma zinc concentration was significantly correlated with sperm density (r = 0.341, P < .0001), motility (r = 0.253, P < .0001), and viability (r = 0.286, P < .0001). On the basis of the findings of this study and those of other reports, zinc may contribute to fertility through its positive effect on spermatogenesis. PMID- 10670520 TI - Spontaneous hyperplasia of the ventral lobe of the prostate in aging genetically hypertensive rats. AB - Recent studies have shown that the prostatic autonomic innervation takes part in its homeostasis and growth. Other works showed that spontaneously hypertensive rats (SHR) show excessive sympathetic activity, accompanied by lower urinary tract symptoms, increased growth capacity of prostatic stromal cells, and increased levels of androgens and their receptors. Furthermore, young SHR were reported to present incipient stages of benign prostatic hyperplasia (BPH). The aim of the present study was to examine whether this strain indeed develops spontaneous BPH with age, and can thus serve as a genuine natural model for this disorder. For this purpose, ventral lobes of prostates of one-year-old, male SHR and their normotensive counterparts, Wistar Kyoto (WKY) rats, were examined histopathologically, and the degree of hyperplasia was evaluated according to a score-chart protocol (histoscore). SHR exhibited severe adenomatous spontaneous BPH, characterized by piling-up of epithelial cells, with papillary formations, accompanied by a mild increase in the amount of fibrocytes and smooth muscle cells in the stroma. This was reflected by histoscore values of 38 +/-2. Thickening of prostatic arterioles also was noted, as well as mild chronic inflammatory exudate. WKY rats did not show any of these features of BPH despite their age (histoscore 17 +/- 3, significantly different from that of SHR). We conclude that SHR can serve as a rodent model for the spontaneous development of BPH with age, most probably due to the excessive neuroendocrine activity characteristic of this rat strain. PMID- 10670521 TI - Correlation of CASA velocity and linearity parameters with sperm mobility phenotype in turkeys. AB - Since all domestic turkeys are produced through artificial insemination, a measurable sperm characteristic that would be predictive of fertility would allow for the culling of poor males, resulting in improved reproductive efficiency. The sperm mobility test (SMT), which quantifies sperm penetration into an Accudenz solution, has been shown to correlate highly with fertilization potential of individual turkeys. Since this sire-selection test is based on the differences in sperm mobility between whole ejaculates from individual males, the objective of this study was to determine whether specific sperm velocity parameters would correlate with the SMT and to determine whether these characteristics could account for phenotypic differences in sperm mobility observed between males. The SMT was used to rank males within a flock (n = 110) in triplicate and to classify them into high, average, and low sperm mobility phenotypes on the basis of the sperm mobility index. Several sperm velocity parameters were evaluated for each male by a computer-aided sperm analysis (CASA) system, the Hobson Sperm Tracker. The types of measurements taken of 200 sperm tracks/ejaculate included the following: curvilinear velocity (VCL), average path velocity (VAP), straight-line velocity (VSL), linearity (LIN), beat-cross frequency (BCF), and mean angular displacement (MAD). Significant positive correlations were found between VSL, LIN, BCF, and sperm mobility, and a significant negative correlation was seen between MAD and sperm mobility. Subpopulations of sperm that had penetrated the Accudenz solution were isolated from each mobility phenotype and were analyzed by CASA, and significant correlations were again observed between VSL, LIN, BCF, and sperm mobility. We conclude that sperm velocity and linearity contribute to overall sperm mobility phenotype and are important characteristics of turkey sperm function. Key words: Motility, computer, spermatozoa. PMID- 10670522 TI - Age-related decrease in hypothalamic gonadotropin-releasing hormone (GnRH) gene expression, but not pituitary responsiveness to GnRH, in the male Brown Norway rat. AB - As is the case in humans, aging male Brown Norway (BN) rats exhibit both primary and secondary (hypothalamic/pituitary) testicular failure. We hypothesized that secondary testicular failure in aging BN rats is due to alterations in both hypothalamic and pituitary function. In order to determine whether gonadotropin releasing hormone (GnRH) gene expression is altered with aging, we compared hypothalamic preproGnRH (ppGnRH) mRNA by in situ hybridization histochemistry and GnRH peptide content in microdissected brain areas by radioimmunoassay in intact (or sham-operated) young, middle-aged, and old male rats. In addition, we determined hypothalamic-pituitary responsiveness to the removal of testicular feedback by comparing ppGnRH messenger RNA (mRNA) and gonadotropin levels in sham operated and orchidectomized young, middle-aged, and old rats. In sham-operated rats, both the cellular ppGnRH mRNA content and the number of neurons expressing ppGnRH mRNA were lower in old compared with young and middle-aged rats. In addition, GnRH content decreased with aging in intact rats in 2 of the 3 brain areas examined, and GnRH content tended to decrease with aging in the third region. Morning serum luteinizing hormone (LH) levels were unchanged with aging, whereas follicle-stimulating hormone (FSH) was significantly increased in old compared with younger intact rats. The cellular ppGnRH mRNA content also decreased with aging in orchidectomized rats, although the number of neurons expressing ppGnRH mRNA was unchanged with aging in these rats. Within age groups, the cellular ppGnRH mRNA content was higher in orchidectomized than in sham operated rats, though there was no effect on the number of neurons expressing GnRH. In a second study, we compared pituitary responsiveness to GnRH by measuring serum LH and FSH levels after GnRH administration in intact BN rats of different ages. The LH response to GnRH was unchanged with aging, whereas the FSH response to GnRH tended to increase with aging. Despite similar LH responses, the testosterone (T) response to GnRH declined progressively with aging. A third study assessed age-related changes in the circadian rhythm of circulating LH, T, and corticosterone (B) levels. LH levels over a 24-hour period decreased with aging and tended to be lower in the morning hours in all age groups, and circadian rhythmicity was blunted in middle-aged and old compared with young rats. T levels over 24 hours declined progressively with aging, and these levels showed a bimodal diurnal variation in young rats, a variation that was not evident in older animals. B levels over a 24-hour period were lower in old than in younger animals, and with aging, there was dampening of the amplitude of the circadian rhythm of B. Taken together, these findings suggest that secondary testicular failure in aging male BN rats is due in part to decreased GnRH gene expression rather than to decreased pituitary responsiveness to GnRH. This reduction in GnRH gene expression with aging is not dependent on testicular feedback factors. Finally, the blunted circadian rhythmicity of LH and T secretion with aging provides further evidence of altered hypothalamic regulation of gonadal hormone secretion in old animals. PMID- 10670523 TI - Transdermal electromotive multi-drug administration for Peyronie's disease: preliminary results. AB - The purpose of this study was to clarify the actual therapeutic potential of a new transdermal drug delivery system (electromotive drug administration; EMDA) for selected patients with Peyronie's disease. Forty patients with Peyronie's disease were treated by electromotive administration of the 3-drug association orgotein-dexamethasone-lidocaine in a double-blind, placebo-controlled, partial crossover study (study 1). Another 25 patients were treated by EMDA with a combination of verapamil-dexamethasone in an uncontrolled study (study 2). Treatment sessions lasted 20 minutes each and took place 3 times a week for 3 weeks with a current of 3 mA. Patients were assessed before treatment and at 1- and 3-month follow-up examinations. Assessments were based on sexual history, physical examination, and dynamic color Doppler ultrasonographic results. Adverse effects of EMDA were not reported. In study 1, the clinical results observed after treatment proved to be significantly better than those of the placebo. Penile pain disappeared in all patients in both studies. Penile lesion (nodule or plaque) either disappeared or significantly improved in 79% and 90% of patients treated by the 3- and 2-drug association, respectively. The improvement of penile deformity also was notable although it did not match the effect observed on penile nodules or plaque (62% and 88%, in studies 1 and 2, respectively). In both studies, more than 80% of patients reported a definite amelioration of penile rigidity, which paralleled the improvement of penile dynamic color Doppler ultrasonographic parameters. Overall, the combination of verapamil-dexamethasone achieved better clinical results than the 3-drug combination. Electromotive drug administration is a novel technique capable of safely achieving satisfactory results in selected patients with Peyronie's disease not only in terms of improvement of patient's symptoms but also due to the reduced need for penile surgery. PMID- 10670524 TI - Androgen receptor gene polymorphism and prostate zonal volumes in Australian and Chinese men. AB - Prostate diseases are age and androgen dependent. The evolution of clinically overt pathology requires decades of exposure to adult male levels of circulating testosterone, but the precise relationship between age and androgen circulation remains poorly understood. A marker of integrated androgen action over prolonged periods would therefore be a valuable tool for clinical and epidemiologic research into the origins of prostate disease. In order to evaluate these 2 factors, we have studied the CAG-repeat length polymorphism of the androgen receptor gene and the size of the total, central, and peripheral zones of the prostate, estimated by planimetric ultrasound in 2 populations with widely different susceptibility to death from invasive prostate cancer. From a larger epidemiologic study of the effects of ethnicity and migration on the origins of prostate disease, a nested-case control study was undertaken with 50 Chinese men living in Yue Yang, China and 50 non-Chinese men living in Sydney, Australia. All men had undergone planimetric transrectal prostate ultrasound together with blood sampling to determine CAG-repeat length by polymerase chain reaction and immunoassay of plasma testosterone, estradiol, dihydrotestosterone (DHT), sex hormone-binding globulin (SHBG), and prostate-specific antigen (PSA). Australian men had larger central (7.9 +/- 0.4 vs 3.3 +/- 0.3 mL) and total (29.8 +/- 1.2 vs 25.5 +/- 1.1 mL) but not peripheral (22.0 +/- 0.9 vs 22.2 +/- 0.8 mL) prostate volumes compared with Chinese men. Even after adjustment for differences in body size (the Australian men were taller and heavier), the central-zone volume remained lower by approximately 50% in Chinese men (P < 0.001), whereas testis and total-prostate volumes were no longer significantly different. The length of CAG repeats was no different between Australian men (22.5 +/- 0.5 repeats) and Chinese men (22.5 +/- 0.5 repeats), and there was no correlation within or between populations in CAG repeats or any measure of prostate volume or hormones. DHT concentration was 20% lower in Chinese men compared with Australian men (1.6 +/- 0.1 vs 2.0 +/- 0.1 nmol/L, P = 0.005), a difference that persisted after age adjustment (P = 0.039) but that was removed by adjustment for differences in total-prostate size (P = 0.12). Blood testosterone, estradiol, SHBG, and PSA concentrations were not different between the 2 populations. Hence, the hypothesis is refuted that the CAG repeat polymorphism in the androgen receptor gene (within the nonpathologic range) and the central-prostate zone volume might be markers of long-term androgen sensitivity. Whether either factor alone may constitute a marker of androgen sensitivity remains to be established by other means, and a long-term marker of integrated androgen action suitable for clinical and epidemiologic research is still lacking. PMID- 10670525 TI - The mouse sperm glycine receptor/chloride channel: cellular localization and involvement in the acrosome reaction initiated by glycine. AB - Previously, we reported that glycine initiates the in vitro acrosome reaction (AR) of porcine and human sperm by a mechanism that includes the glycine receptor/Cl- channel (GlyR) and that this receptor/channel is required for the zona-pellucida-initiated AR. Because mouse sperm are important tools in the study of fertilization, we investigated whether glycine initiated the mouse sperm AR and whether the sperm GlyR was involved in that initiation. Glycine (250 microM to 1 mM) initiated the AR of capacitated but not noncapacitated mouse sperm. The glycine-initiated AR was significantly inhibited by 50 nM strychnine, a neuronal GlyR antagonist. The neuronal GlyR agonists taurine and beta-alanine did not initiate the AR at 1 mM or 5 mM. A monoclonal antibody against the rat spinal cord GlyR significantly inhibited the glycine-initiated AR but not the spontaneous AR. Indirect immunofluorescence localization studies with that monoclonal antibody and postfixed live sperm detected 3 patterns of immunoreactivity involving 2 sites in the periacrosomal plasma membrane. These patterns were as follows: type A localization on the plasma membrane overlying the tip of the anterior acrosomal region; type B localization on the plasma membrane overlying the posterior part of the acrosomal equatorial segment and/or, in acrosome-reacted sperm, the posterior part of the modified equatorial segment; and type C localization that included both type A and type B. Type A and type C localization were only observed on the acrosome-intact sperm. During capacitation, the number of the sperm showing type A localization increased. Our results demonstrate that mouse sperm provide an excellent model for studying the role of the GlyR in the acrosome reaction. PMID- 10670526 TI - Morphologic changes in efferent ductules and epididymis in estrogen receptor alpha knockout mice. AB - Estrogen has been shown to have an important role in fluid reabsorption in efferent ductules of the testis. Our previous study of the estrogen receptor alpha knockout mouse (ERKO) showed that the efferent ductules and rete testis were primary targets of estrogen receptor function. In the present study, a more comprehensive evaluation of the ERKO male reproductive tract was performed to determine the severity of effects in efferent ductules as well as the epididymis. The following observations were found in ERKO males: 1) blind-ending efferent ductules were more prevalent in ERKO than in wild type (WT) tissues; 2) glycogen containing cells were observed at the rete testis-efferent ductule junction; 3) the tubular diameters of the efferent ductules and initial segment epididymides were dilated; 4) efferent ductules were dilated between 130 to 300% over wild type ductules; 5) efferent ductule epithelial height was reduced nearly 50%; 6) microvilli of nonciliated cells of efferent ductules were 64% shorter in length; 7) cilia were reduced in number; 8) initial segment epithelium was displaced into regions adjacent to the rete testis and in short segments of the common region of efferent ductule; 9) apical, narrow, and clear cells of the epididymis also were abnormal in some regions; 10) in the corpus and cauda regions, sperm granulomas were noted in one third of the ERKO males. In conclusion, the entire reproductive tract is affected in ERKO males. The cells showing the greatest effects were estrogen receptor-positive cells. It appears that in the ERKO mouse there are developmental anomalies that must be considered separately from adult dysfunctional changes in the male reproductive tract. PMID- 10670527 TI - Circulating and luminal testicular factors affect LRP-2 and Apo J expression in the epididymis following efferent duct ligation. AB - Apolipoprotein J (clusterin or sulfated glycoprotein-2) has been shown to be secreted by the epididymal principal cells, whereupon it binds to sperm in the lumen. Apolipoprotein J also is endocytosed by principal cells along the epididymis. Recently, it has been demonstrated that low-density lipoprotein receptor-related protein-2 (LRP-2) mediates the endocytosis of Apo J and is present in the epididymis. The purpose of the present study was to determine the factors regulating the synthesis of these 2 proteins in various experimentally treated animals. The epididymides of adult rats were fixed with Bouin's fluid and examined with anti-Apo J and anti-LRP-2 antibodies by a light microscope immunocytochemical method. In normal adult animals, expression of Apo J was evident in principal cells of all epididymal regions except the proximal initial segment. Diffuse cytoplasmic staining indicated Apo J secretion. Reactive apical vesicles, presumably endosomal in nature, suggested endocytosis of Apo J. Lipoprotein receptor-related protein-2 expression was solely apical in nature and was seen as an intense apical band in principal cells of all regions except the proximal and distal initial segment and distal caput regions of the epididymis. Hypophysectomy, up to 28 days after the procedure, did not affect expression of Apo J or LRP-2 in principal cells along the entire epididymis. Orchidectomy, with or without testosterone replacement at all time intervals examined, also did not affect LRP-2 expression along the entire epididymis. This also was noted for Apo J expression in all regions except the proximal initial segment. Thus, expression of these 2 proteins does not appear to be regulated by testicular or pituitary factors. In contrast, bilateral as well as unilateral (intact and ligated sides) efferent duct ligation resulted in dramatic differences in LRP-2 and Apo J expression in principal cells in the various epididymal regions. In the case of LRP-2, a complete absence of reaction was noted in principal cells along the entire epididymis. As for Apo J, expression in the distal initial segment, intermediate zone, and caput region remained unchanged compared with that in normal adult animals, whereas in the corpus and cauda epididymides, results of cytoplasmic staining were negligible. These results suggest that under conditions of efferent duct ligation, a circulating factor emanates from the testis to inhibit expression of LRP-2 and Apo J in these epididymal regions. Furthermore, because Apo J was affected in a region-specific manner, unlike the case for LRP 2, different factors appear to be involved for each protein. These factors may be produced to inhibit proteins from being synthesized by the epididymis in the absence of luminal testicular input and may exist in cases of congenital and pathologic epididymal tubule blockages as well as after vasectomy. In the case of immunostaining for Apo J in the proximal initial segment only, normally unreactive principal cells in control adult animals became intensely reactive after orchidectomy as well as bilateral and unilateral (ligated side only) ligation. As this was not the case for hypophysectomized animals and the intact side of unilateral efferent duct-ligated animals, it is suggested that a testicular factor entering via the lumen of the efferent ducts serves to inhibit Apo J expression in this area. The present data also reveal that after efferent duct ligation, there are circulating factors that inhibit Apo J expression in a region-specific manner (corpus and cauda) and that inhibit LRP-2 expression along the entire epididymis and that these are derived from the testis. Furthermore, the data reveal that a testicular luminal factor appears to inhibit Apo J expression in the proximal initial segment of normal adult animals. Key words: Principal cells, orchidectomy, glycoprotein 330, clusterin, sulfated glycoprotein 2. PMID- 10670528 TI - Semen quality and human fertility: a prospective study with healthy couples. AB - Measures of semen quality are used as surrogate measures of male fertility in clinical andrology, reproductive toxicology, epidemiology, and risk assessment. However, only limited data are available to relate those measures to fertility. This prospective study with 210 reproductive-age couples was conducted to provide information on the value of semen quality measures for predicting human male fertility potential and for development of models to estimate the effects of changes in semen quality on fertility in a given population for risk assessment. Couples without known risk factors for infertility and who had discontinued contraception to have a child were accepted. The study followed each couple for up to 12 menstrual cycles while they attempted to conceive and evaluated semen quality measures from multiple ejaculates per man with known abstinence intervals. For each cycle, the day of ovulation was predicted, and the couple was advised to have intercourse multiple times on that day and on the days around it. Among the demographic variables assessed, parity, contraception status prior to entering the study, male education level, and male smoking were associated significantly with 12-cycle pregnancy rate. Several semen quality measures also were associated significantly with pregnancy rate, with percentage morphologically normal sperm by strict criteria and measures involving total number of sperm showing particularly strong associations. Localized regression smoothing plots of semen quality data against proportion of couples pregnant suggested levels below which fertility declines for several semen quality measures. These results have applications in both clinical andrology and in assessment of risk to male fecundity from environmental or pharmaceutical exposures. In particular, they contribute information on behavior of fertility with varying semen quality and can allow development of models to predict effects on fertility in populations from decrements in semen quality. PMID- 10670530 TI - Establishment of the Male Reproductive Biology Club. PMID- 10670529 TI - Maturation of epididymal spermatozoa in the nondomesticated guinea pigs Cavia aperea and Galea musteloides. AB - The physiological changes occurring in spermatozoa in the male reproductive tract of 2 nondomesticated species of South American guinea pigs with different mating systems were studied. Cavia aperea, the wild ancestor of the domesticated guinea pig, has a polygynous mating system, whereas Galea musteloides exhibits promiscuous mating behavior. The epididymis of both species resembled that of the domesticated guinea pig, with a swathe of tubule convolutions (linking the 2 major parts of the organ) that was of smaller size in Cavia but not Galea. Higher relative epididymal weight was demonstrated in the promiscuous species. During their journey through the epididymis, spermatozoa from Galea developed their potential for motility expression more proximally than did those of Cavia, but motility developed into forward progression in the same region in both species. The maximal velocities exhibited by mature Cavia sperm in vitro were greater than those of Galea. Spermatozoa from Cavia were twice the length of those from Galea, they had larger heads, and the acrosomes of single sperm were more sensitive to disruption during morphological preparation. Only in Cavia did agglutination of sperm into rouleaux occur, after the potential for motility had been developed. Migration of the cytoplasmic droplet along the midpiece occurred in the same regions in both species and before agglutination in Cavia. It is suggested that the male's reproductive strategy (polygyny vs promiscuity) dictates the size of the testis and epididymis, whereas the female's reproductive physiology (induced ovulation vs cyclicity) influences the posttesticular development of sperm morphology and motility in the epididymis. PMID- 10670531 TI - The American Society of Andrology. PMID- 10670532 TI - Over population is the biggest problem faced by our planet. PMID- 10670533 TI - Andrology as a discipline. PMID- 10670534 TI - The future for andrology. PMID- 10670535 TI - The future of andrology in the next millennium. PMID- 10670536 TI - The future of andrology. PMID- 10670537 TI - What may transpire in the field of andrology in the early part of the next century. PMID- 10670538 TI - The future of ASA. American Society of Andrology. PMID- 10670539 TI - The future of andrology is bright. PMID- 10670541 TI - The future of andrology. PMID- 10670540 TI - The future of andrology. PMID- 10670542 TI - The future of andrology. PMID- 10670544 TI - The future of andrology. PMID- 10670543 TI - Andrology at the time of the second millennium. PMID- 10670545 TI - Risk factors for human herpesvirus 8 seropositivity and seroconversion in a cohort of homosexual men. AB - Sexual and nonsexual modes of transmission of human herpesvirus 8 (HHV8) have been suggested, but specific routes remain unclear. Therefore, the objective of this study was to assess risk factors for HHV8 seropositivity and determine specific sexual practices associated with HHV8 seroconversion. Sera from 1,458 homosexual men (Amsterdam Cohort Study, 1984-1996) were tested for antibodies to HHV8 with a modified version of an enzyme immunoassay, using recombinant HHV8 lytic phase capsid (ORF65) and latent phase nuclear (ORF73) proteins. HHV8 seroprevalence at study entry was 20.9% (305/1,458); was highest among those with positive human immunodeficiency virus (HIV) status, no steady partner, and southern European or Latin American nationality; and increased with older age and higher number of sexual partners. During follow-up, 215 men seroconverted for HHV8 (incidence: 3.6/100 person-years). Both prevalence and incidence rates remained more or less stable during the study period. Orogenital insertive sex (odds ratio (OR) = 5.95; 95% confidence interval (CI): 2.88, 12.29) or orogenital receptive sex (OR = 4.29; 95% CI: 2.11, 8.71) with more than five partners in the past 6 months, older age (OR = 2.89; 95% CI: 1.13, 7.34, when older than 45 years), and preceding HIV infection (OR = 2.47; 95% CI: 1.53, 3.99) were independent predictors for HHV8 seroconversion. The authors found strong evidence for orogenital transmission of HHV8 among homosexual men. PMID- 10670546 TI - Invited commentary: determining specific sexual practices associated with human herpesvirus 8 transmission. AB - Laboratory and epidemiologic studies have established human herpesvirus 8 (HHV8) as an etiologic agent of Kaposi's sarcoma. With strong evidence linking HHV8 infection with the number of sexual partners among homosexual men, the challenge now is to determine the specific sexual acts associated with HHV8 transmission. Initial studies of specific practices, however, have differed in their conclusions; the paper by Dukers et al. in this issue of the Journal is the first to associate penile-oral intercourse with HHV8 transmission. Many sources of bias may contribute to the conflicting findings of studies reported to date: HHV8 research still lacks an adequately specific and sensitive serologic assay; identification of relevant exposure periods and measurement of sexual practices are imperfect; and sufficient adjustment for confounding is problematic. These numerous potential biases may be particularly important when trying to detect underlying associations that may be of low-order magnitude. The study by Dukers et al. (Am J Epidemiol 2000;151:213-24) is an important contribution to research on HHV8 transmission, but we do not yet know enough about the possible sexual routes of transmission to recommend avoiding any single behavior. For now, the best prevention advice is to reinforce the more general safe sex practices that have been promoted to prevent human immunodeficiency virus and other sexually transmitted diseases. PMID- 10670547 TI - Paternal military service and risk for childhood leukemia in offspring. AB - To assess the association between paternal military service and risk for childhood leukemia, the authors analyzed data from three case-control studies conducted by the Children's Cancer Group from 1983 to 1993. A total of 605 acute myeloid leukemia (AML, age < or = 18 years) cases, 2,117 acute lymphoblastic leukemia (ALL, age < or = 14 years) cases, and 3,155 individually matched controls were included in these studies. Paternal military history and other exposure data were obtained in 2,343 matched case-control sets, including 1,805 ALL and 528 AML cases. Paternal general military service was not associated with the leukemia risk. A small, but significant, increase in the risk for AML was seen, however, among offspring of veterans who had served in Vietnam or Cambodia (odds ratio (OR) = 1.7; 95% confidence interval (CI): 1.0, 2.9), after adjustment for paternal education, race, income, smoking, X-ray exposure, and marijuana use. The risk was predominantly present in children diagnosed before the age of 2 (OR = 4.6; 95% CI: 1.3, 16.1), although there were inconsistencies in the risks associated with length of time served and interval between service and diagnosis of leukemia. Military service in Vietnam or Cambodia was unrelated to the risk for ALL. The etiologic importance, if any, of these observations has yet to be determined. PMID- 10670548 TI - Environmental tobacco smoke and lung cancer: a case-control study in Germany. AB - To assess the association between exposure to environmental tobacco smoke (ETS) and lung cancer, the authors personally interviewed 292 lifelong nonsmoking lung cancer cases (recruited from 15 hospitals in the study area) and 1,338 nonsmoking controls (randomly selected by population registries) between 1990 and 1996 in Germany. Subjects were asked by a standardized questionnaire about exposure to ETS in childhood, by spouse, at work, and in transportation and social settings. Several indicators of these different sources of exposure were investigated, using not or low exposed subjects as the reference category. The most informative quantification index was weighted duration of exposure (hours x level of smokiness). No effect of ETS exposure during childhood and no clear effect of spousal ETS were observed. However, for the highest category of exposure, clear effects of ETS at the workplace (odds ratio (OR) = 1.93; 95% confidence interval (CI): 1.04, 3.58), in vehicles (OR = 2.64; 95% CI: 1.30, 5.36), and from all sources combined (OR = 1.39; 95% CI: 0.96, 2.01) were found. Adjustment for occupational carcinogens, radon, and diet did not appreciably change the results. These findings suggest that exposures to high levels of ETS at the workplace and in other public indoor settings appear to be important risk factors for lung cancer risk in nonsmokers. PMID- 10670549 TI - Saliva cotinine levels in smokers and nonsmokers. AB - The authors collected by mail self-reported data on smoking habits and saliva samples that were analyzed for cotinine concentration in 222 smokers and 97 nonsmokers. Participants were members of the University of Geneva (Switzerland) in 1995. The 207 cigarette-only smokers smoked on average 10.7 cigarettes/day and had a median concentration of cotinine of 113 ng/ml. The cotinine concentration was moderately associated with the number of cigarettes smoked per day (+14 ng/ml per additional cigarette, p < 0.001, R2 = 0.45) and was 54 ng/ml higher in men than in women after adjustment for cigarettes per day and for the Fagerstrom Test for Nicotine Dependence. The cotinine level was not associated with the nicotine yield of cigarettes (r= 0.08). In nonsmokers, the median concentration of cotinine was 2.4 ng/ml. The cotinine concentration was 1.5 times higher in nonsmokers whose close friends/spouses were smokers than in nonsmokers whose close friends/spouses were nonsmokers (p = 0.05). A cutoff of 7 ng/ml of cotinine distinguished smokers from nonsmokers with a sensitivity of 92.3% and a specificity of 89.7%; a cutoff of 13 ng/ml provided equally satisfactory results (sensitivity, 86.5%; specificity, 95.9%). This study provides evidence for the construct validity of both questionnaires and saliva cotinine for the assessment of active and passive exposure to tobacco smoke. PMID- 10670550 TI - Environmental exposure to tremolite and respiratory cancer in New Caledonia: a case-control study. AB - A case-control study on respiratory cancers was conducted in New Caledonia (South Pacific), where a high incidence of malignant pleural mesothelioma had been observed. The disease pattern suggested an environmental exposure to asbestos. The first results showed that, in some areas, tremolite asbestos derived from local outcroppings was used as whitewash (locally named "po"). All cases diagnosed between 1993 and 1995 (including 15 pleural mesotheliomas, 228 lung cancers, and 23 laryngeal cancers) and 305 controls were included in the study. Detailed information on past or present use of the whitewash, residential history, smoking, diet, and occupation was collected. The risk of mesothelioma was strongly associated with the use of the whitewash (odds ratio (OR) = 40.9; 95% confidence interval (CI): 5.15, 325). All Melanesian cases had been exposed. Among Melanesian women, exposure to the whitewash was associated with an increased risk of lung cancer (OR = 4.89; 95% CI: 1.13, 21.2), and smokers exposed to po had an approximately ninefold risk (OR = 9.26; 95% CI: 1.72, 49.7) compared with women who never smoked and had never used the whitewash. In contrast, no association was noted between exposure to po and lung cancer risk among Melanesian men, probably because of lower exposure levels. Among non Melanesians, the numbers of exposed subjects were too small to assess the effect of exposure to po. There was no indication of elevated risks for the other cancer sites. PMID- 10670551 TI - An iatrogenic epidemic of benign meningioma. AB - Head irradiation, the acceptable mode of treatment for tinea capitis in the past, is recognized today as a causative factor for meningioma. This treatment was applied en mass to immigrants coming to Israel from North Africa and the Middle East during the 1950s. In order to estimate the effect of the differential radiation treatment on the rates of meningioma in the total population, the authors assessed time trends of this disease in Israel over the past 40 years by main ethnic origin. Cohort analysis shows a marked incidence rise in the North African-born cohorts born in 1940-1954 starting from the 1980s. A similar pattern is seen in the Middle Eastern born, although the increase is not as sharp. In consequence, there is a crossover of the interethnic incidence curves in the 1940 1949 cohort. Comparison of the relative risk between 1940-1954 cohorts that comprised most of the irradiated with 1930-1939 cohorts, who were largely free of the radiation, shows that the North African born have the largest relative risk of 4.62, followed by the Middle Eastern born, with a relative risk of 1.95, while the European-American born have a relative risk close to 1. The differences between the three areas of birth are statistically significant. The data illustrate the potential risk of administering highly potent therapy for an essentially benign disease that led, in turn, to a drastic change in the national meningioma pattern. PMID- 10670552 TI - Examination of the relation between periodontal health status and cardiovascular risk factors: serum total and high density lipoprotein cholesterol, C-reactive protein, and plasma fibrinogen. AB - Using data from the Third National Health and Nutrition Examination Survey (1988 1994), the authors examined the relation between periodontal health and cardiovascular risk factors: serum total and high density lipoprotein cholesterol, C-reactive protein, and plasma fibrinogen. A total of 10,146 participants were included in the analyses of cholesterol and C-reactive protein and 4,461 in the analyses of fibrinogen. Periodontal health indicators included the gingival bleeding index, calculus index, and periodontal disease status (defined by pocket depth and attachment loss). While cholesterol and fibrinogen were analyzed as continuous variables, C-reactive protein was dichotomized into two levels. The results show a significant relation between indicators of poor periodontal status and increased C-reactive protein and fibrinogen. The association between periodontal status and total cholesterol level is much weaker. No consistent association between periodontal status and high density lipoprotein cholesterol was detectable. Similar patterns of association were observed for participants aged 17-54 years and those 55 years and older. In conclusion, this study suggests that total cholesterol, C-reactive protein, and fibrinogen are possible intermediate factors that may link periodontal disease to elevated cardiovascular risk. PMID- 10670553 TI - Social patterning of myocardial infarction and stroke in Sweden: incidence and survival. AB - Cardiovascular disease morbidity and mortality rates show marked social patterning in industrialized countries. The aim of this study was to analyze if not only incidence but also survival after acute myocardial infarction (AMI) and stroke differ among socioeconomic groups. Within the framework of the population based World Health Organization's Multinational Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA) Project, all first-ever AMI (ages 25-64 years) and stroke (ages 25-74 years) events were recorded in northern Sweden during the period 1985-1994. The numbers of first-ever AMI and stroke patients included in the study were 3,466 and 4,215, respectively. Incidence rates for both AMI and stroke showed a distinct social pattern, with high rates in workers and self-employed nonprofessionals and low rates in professionals. The pattern was similar in men and women. In men, early survival after an AMI follows the same socioeconomic pattern, whereas it is less clear if socioeconomic differences in survival contribute to explain differences in mortality in AMI among women and mortality in stroke (both sexes). The high case fatality among male workers and self-employed professionals with AMI is, in turn, attributed to a very marked increase in the risk for sudden death. PMID- 10670554 TI - Associations of light, moderate, and vigorous intensity physical activity with longevity. The Harvard Alumni Health Study. AB - Physical activity is associated with better health; however, the optimal intensity of activity remains unclear. A total of 13,485 men (mean age, 57.5 years) from the Harvard Alumni Health Study reported their walking, stair climbing, and sports/recreation in 1977. Between 1977 and 1992, 2,539 died. After adjusting for the different activity components, distance walked and storeys climbed independently predicted longevity (p, trend = 0.004 and <0.001, respectively). Light activities (<4 multiples of resting metabolic rate (METs)) were not associated with reduced mortality rates, moderate activities (4-<6 METs) appeared somewhat beneficial, and vigorous activities (> or =6 METs) clearly predicted lower mortality rates (p, trend = 0.72, 0.07, and <0.001, respectively). These data provide some support for current recommendations that emphasize moderate intensity activity; they also clearly indicate a benefit of vigorous activity. PMID- 10670555 TI - Deaths attributable to childbearing in Matlab, Bangladesh: indirect causes of maternal mortality questioned. AB - Little is known about the nature of diseases aggravated by pregnancy or the magnitude of mortality from causes indirectly related to pregnancy. This study aims at clarifying the contribution of indirect causes to maternal mortality by analyzing the problem from an epidemiologic perspective, using population-based data from Matlab, Bangladesh, for the period 1976-1993. The time spent during pregnancy and the puerperium was considered a transitory exposure period in women's lives, and death rates were calculated for women aged 15-44 years, while exposed and while not exposed. During or shortly after pregnancy, death rates from all causes are more than twice as high as outside this period. Once direct obstetric causes and injuries are excluded, the death rates among women while exposed are substantially lower than the death rates among women while not exposed. Several interpretations of this finding are discussed, particularly the role of selective factors ("healthy pregnant woman effect"?). This study highlights the complexity of the concept of indirect causes of maternal mortality and clearly illustrates the inherent difficulties in estimating the excess risk of death attached to pregnancy and the puerperium. PMID- 10670556 TI - US Department of Veterans Affairs medical care system as a resource to epidemiologists. AB - Epidemiologists have utilized several health care systems with large numbers of enrollees and centralized databases to achieve their research aims. Although containing many of the features that have made certain health care systems valuable to the conduct of epidemiologic research, the US Department of Veterans Affairs (VA) medical care system has not been well utilized by epidemiologists. This article will describe existing and planned features of this health care system that should be of interest to epidemiologists, including centralized databases that capture hospital discharge and outpatient clinic diagnostic data, a planned enrollment file that would contain all persons eligible for VA medical care, and the size and national dispersion of VA medical care facilities. Also, VA leadership has demonstrated an interest in the promotion of epidemiologic research by initiating several new programs, including the creation of three Epidemiologic Research and Information Centers (ERICs) to foster VA epidemiologic research, and announcing a program to support investigator-initiated epidemiologic research projects with VA funding. Epidemiologists with interests in medical problems that afflict veterans should consider partnerships with VA investigators to achieve their research aims. PMID- 10670557 TI - Do health interview surveys yield reliable data on chronic illness among older respondents? AB - Previous research evaluating quality of health interview survey data has generally relied upon comparisons of household interview data with medical records or other external sources of information. However, "gold standards" are not always satisfactory or available. This paper illustrates an alternative approach to the evaluation of data quality-examination of the reliability of reports of chronic conditions in longitudinal surveys. The data come from national samples of older Americans (First National Health and Nutrition Examination Survey Epidemiologic Followup Study, 1971-1975, 1982-1984, 1986) and older Taiwanese (Survey of Health and Living Status of the Elderly, 1989, 1993, 1996). The results show that, among respondents who reported a chronic condition at a given interview, the likelihood that the condition was acknowledged at the subsequent interview was higher for hypertension and diabetes than for arthritis and stroke. Low levels of consistency for stroke appear to result partly from the poor wording of questions. In Taiwan, younger, more educated persons and those experiencing severe conditions were somewhat more likely to acknowledge the condition at follow-up compared with their respective counterparts. Women and persons of high cognitive status in the United States and respondents in both countries who used a proxy to report the occurrence of a stroke were also more likely to acknowledge conditions at follow-up. PMID- 10670558 TI - Genetic distances for the study of infectious disease epidemiology. AB - Molecular epidemiologic studies of infectious pathogens 1) generate genetic patterns from a collection of microorganisms, 2) compare the degree of similarity among these patterns, and 3) infer from these similarities infectious disease transmission patterns. The authors propose a quantitative approach using genetic distances to study the degree of similarity between patterns. Benefits of such genetic distance calculations are illustrated by an analysis of standard DNA fingerprints of Mycobacterium tuberculosis in San Francisco collected during the period 1991-1997. Graphical representation of genetic distances can assist in determining if the disappearance of a specific pattern in a community is due to interruption of transmission or ongoing evolution of the microorganism's fingerprint. Genetic distances can also compensate for varying information content derived by DNA fingerprints of contrasting pattern complexity. To study demographic and clinical correlates of transmission, the authors calculated the smallest genetic distance from each patient sample to all other samples. With correlation of genetic distances and nearest genetic distances with previously understood notions of the epidemiology of M. tuberculosis in San Francisco, factors influencing transmission are investigated. PMID- 10670559 TI - Re: "Biased tests of association: comparisons of allele frequencies when departing from Hardy-Weinberg proportions". PMID- 10670560 TI - Reducing the use of antimicrobial agents in animals and man. PMID- 10670561 TI - Expression of putative virulence factors by clinical isolates of Klebsiella planticola. AB - A total of 92 clinical isolates of Klebsiella planticola from man was examined with respect to the production of haemagglutinins and siderophores, serum resistance and distribution of capsular types. For comparison, a group of 207 clinical isolates of K. pneumoniae was also studied. The percentages of K. planticola strains able to express mannose-sensitive haemagglutination, indicating type 1 fimbriae (83%) and mannose-resistant and Klebsiella-like agglutination, indicating type 3 fimbriae (69%), as well as to produce the siderophores enterobactin (100%) and aerobactin (2.2%) were almost identical to those of the K. pneumoniae strains. Similarly, the proportion of serum-resistant strains (30%) was comparable to that of K. pneumoniae (25%). The capsule types most often detected in K. planticola were K14 (13%), K2 (9%) and K70 (9%). The incidence of K2, which is the predominant capsular type in K. pneumoniae, was similar in both species. These findings show that K. planticola, which is being detected with increasing frequency in clinical specimens from man, has the ability to express similar putative virulence factors to K. pneumoniae, suggesting that they may have similar pathogenicity. PMID- 10670562 TI - Production of an enterotoxin by a gastro-enteritis-associated Aeromonas strain. AB - The potential of motile Aeromonas species to cause human gastrointestinal infections has been recognised recently. Considerable worldwide epidemiological, microbiological and clinical investigations have shown that some strains of the different motile aeromonads are of increasing enteropathogenic significance, especially in children, the elderly and in immunocompromised individuals. Some of the diarrhoeal symptoms of Aeromonas-associated gastro-enteritis have been attributed to enterotoxins. In this study, 15 Aeromonas isolates from clinical and non-clinical sources, representing the three motile aeromonads commonly associated with gastro-enteritis (A. caviae, A. hydrophila and A. veronii biovar sobria), were tested for their ability to cause fluid accumulation in infant mice by the suckling mouse technique. Eight isolates were found to produce enterotoxin. Of these, an A. veronii biovar sobria strain (AS15), isolated from lamb kidney, was found to produce the highest enterotoxin score. An enterotoxin of c. 40 kDa produced by A. veronii biovar sobria AS15 was purified by Sephacryl S-100 gel filtration and high-performance liquid chromatography. This enterotoxin caused marked fluid accumulation in infant mice by the suckling mouse technique. The purified enterotoxin cross-reacted with cholera toxin antibodies and was readily inactivated by heating at 56 degrees C for 10 min. The production of a 'cholera-like' enterotoxin by Aeromonas isolates from samples of animal origin suggests that these organisms could be of public health significance in food products. PMID- 10670563 TI - Biological activities of lipopolysaccharides of Proteus spp. and their interactions with polymyxin B and an 18-kDa cationic antimicrobial protein (CAP18)-derived peptide. AB - The saccharide constituents of lipopolysaccharides (LPS) of Proteus spp. vary with the strain and contain unique components about which little is known. The biological activities of LPS and lipid A from S- and R-forms of 10 Proteus strains were examined. LPS from all S-form Proteus strains was lethal to D-(+) galactosamine (GalN)-loaded, LPS-responsive, C3H/HeN mice, but not to LPS-hypo responsive C3H/HeJ mice. P. vulgaris 025 LPS evoked strong anaphylactoid reactions in N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP)-primed C3H/HeJ mice. LPS from S- and R-form Proteus strains induced production of nitric oxide (NO) and tumour necrosis factor (TNF) by macrophages isolated from C3H/HeN but not C3H/HeJ mice. Lipid A from Proteus strains also induced NO and TNF production, although lipid A was less potent than LPS. The effects of LPS were mainly dependent on CD14; LPS-induced NO and TNF production in CD14+ J774.1 cells was significantly greater than in CD14-J7.DEF.3 cells. All LPS from Proteus strains, and especially from P. vulgaris 025, exhibited higher anti-complementary activity than LPS from Escherichia coli or Pseudomonas aeruginosa. Polymyxin B inactivated proteus LPS in a dose-dependent manner, but these LPS preparations were more resistant to polymyxin B than E. coli LPS. CAP18(109-135), a granulocyte-derived peptide, inhibited proteus LPS endotoxicity only when the LPS:CAP18(109-135) ratio was appropriate, which suggests that CAP18(109-135) acts through a different mechanism than polymyxin B. The results indicate that LPS from Proteus spp. are potently endotoxic, but that the toxicity is different from that of LPS from E. coli or Salmonella spp. and even varies among different Proteus strains. The variation in biological activities among proteus LPS may be due to unique components within the respective LPS. PMID- 10670564 TI - Ultrastructural study of Mycobacterium avium infection of HT-29 human intestinal epithelial cells. AB - Mycobacterium avium is a common pathogen in AIDS patients and, in a large percentage of those patients, M. avium infection appears to be acquired via the gastrointestinal tract. M. avium is able to bind to and enter human and murine intestinal epithelial cells in vitro and in vivo. The invasion by and intracellular fate of M. avium in the HT-29 intestinal epithelial cell line was examined in an ultrastructural study. Bacterial contact with polarised cells was observed 10-15 min after monolayer infection and in polarised monolayers this always occurred in areas lacking microvilli. Contact with HT-29 cells did not appear to take place in a preferential area on the bacterial cell. Following invasion, M. avium was encountered within vacuoles containing either single or multiple bacteria; the latter evolved to contain only an individual bacterium. Vacuoles containing more than one bacterium were seen early in the infection and eventually underwent segmentation, with each bacterium occupying a vacuole. No bacteria were observed outside vacuoles up to 5 days after infection. PMID- 10670565 TI - Development and evaluation of a solid-phase enzyme immunoassay based on Andes hantavirus recombinant nucleoprotein. AB - Hantavirus pulmonary syndrome (HPS) with high mortality rate has been reported in five countries in South America. Rapid accurate methods are important both for monitoring acute infections and for epidemiological studies. The Andes virus nucleoprotein amino acid sequence has a high identity percentage compared with other sequences of this region and has been chosen for the development of diagnostic reagents. Andes nucleoprotein expressed in Escherichia coli was applied as antigen in IgG, IgA and mu-capture IgM enzyme-linked inmunosorbent assays (ELISAs). An evaluation of this reagent was conducted to establish its usefulness for differential diagnosis of HPS and seroprevalence studies. Samples from 135 reverse transcription (RT)-PCR-confirmed HPS cases, 77 individuals with other respiratory infections and 957 healthy inhabitants from endemic and non endemic areas were analysed. The hantavirus-infected patients had an early and strong IgM, IgG and IgA serum antibody response, in most of the cases as early as 1, 7 and 1 days following onset of symptoms, respectively. IgM and IgG detection showed a specificity and sensitivity of 100%. Andes-specific IgM antibodies were found in all patients in the first available sample, which remained detectable for at least 43 days. Specific IgA antibodies were also detected in saliva of patients with acute HPS. The short duration of the disease and the risk for contacts due to person-to-person transmission of Andes virus necessitate the use of highly sensitive tests which might lead to earlier detection of infected people and improve the treatment and management of patients with HPS. PMID- 10670567 TI - Identification and partial characterisation of a new protective antigen of Brucella abortus. AB - Two novel Brucella abortus proteins were isolated from B. abortus strain RB51 and their immunological properties were determined. These proteins precipitated in the 40-60% saturated concentration range of ammonium sulphate and had a molecular mass of 32.2 kDa and 22.9 kDa, respectively. Both were able to induce a strong in vitro blast transformation in lymphoid cells obtained from mice previously sensitised with a crude brucella protein extract. The protection studies showed that the 22.9-kDa protein used as a protective immunogen was as effective as the live B. abortus RB51 vaccine but the 32.2-kDa protein had a poor protective effect under similar conditions. The amino-terminal sequence of the 22.9-kDa and 32.2-kDa proteins was determined and analysed in a database. The lack of homology with other known B. abortus proteins indicated that both proteins were novel antigens. PMID- 10670566 TI - A nasal whole-cell pertussis vaccine induces specific systemic and cross-reactive mucosal antibody responses in human volunteers. AB - A whole-cell pertussis vaccine, each dose consisting of 250 microg of protein, was given intranasally four times at weekly intervals to six adult volunteers. All vaccinees responded with increases in nasal fluid IgA antibodies to Bordetella pertussis whole-cell antigen. Three vaccinees with high nasal antibody responses also developed increased serum IgA and IgG antibodies to this antigen. Salivary antibody responses to the whole-cell antigen, as well as antibodies in serum and secretions to pertussis toxin (PT) and filamentous haemagglutinin (FHA) were negligible, except for a moderate increase in nasal fluid antibodies to FHA. Unexpectedly, the same vaccinees developed significant rises in nasal and salivary IgA antibodies to meningococcal outer-membrane antigens, whereas corresponding serum IgA and IgG antibodies were unchanged. Thus it appears that mucosal immunisation may induce secretory antibodies with broader specificities than can be found in serum. PMID- 10670568 TI - Role of interleukin-6 in determining the course of murine Tyzzer's disease. AB - Clostridium piliforme is an obligately intracellular bacterium that causes enterohepatic disease in many domestic and laboratory animal species. Susceptibility to infection is known to vary with the host immune status, species and strain, but little is known about specific immune mechanisms that regulate this disease. Subclinical infection was induced in weanling C. piliforme susceptible DBA/2 or resistant C57BL/6 mice with either a toxic or a non-toxic C. piliforme isolate. Hepatic lesions and bacteria were evident in both mouse strains for 14 days after inoculation with the toxigenic bacterial isolate, but were never demonstrated following inoculation with the non-toxigenic isolate. All mice demonstrated increased interleukin-6 (IL-6) levels that were largely independent of host strain susceptibility to infection or virulence of the bacterial isolate. The severity of C. piliforme-induced hepatic lesions was increased by polyclonal anti-IL-6 treatment in both resistant (DBA/2) and susceptible (C57BL/6) mouse strains. These data indicate that IL-6 is important in mediating the course of murine C. piliforme infections but is not involved in determining host susceptibility to acute infection, nor is it influenced by the virulence of the C. piliforme isolate. PMID- 10670569 TI - Clonal groups of enteropathogenic Escherichia coli isolated in case-control studies of diarrhoea in Bangladesh. AB - Recent case-control studies in Bangladesh showed a high prevalence of enteropathogenic Escherichia coli (EPEC) strains (identified by DNA probes for virulence genes) associated with childhood diarrhoea. However, the clonal status of these strains is not known. A total of 94 EPEC isolates from 80 children with diarrhoea and 14 healthy matched controls isolated during 1991-1992 and 1993-1994 was characterised by serogrouping, enterobacterial repetitive intergenic consensus sequence PCR, and by a biochemical fingerprinting method (the phene plate or PhP system). Twelve O serogroups were found with O114 (n = 19) and O127 (n = 23) being the dominant serogroups. Most strains of O114 belonged to the same PhP/PCR types. Strains of O127 contained 16 that produced cytolethal distending toxin (CDT) and seven that did not; both were found among patients as well as controls. Results of PCR and PhP typing showed that CDT-positive strains belonged to the same clonal group and were related to one of the two PhP/PCR types of CDT negative O127 strains. Thirty-one EPEC strains were O non-typable and 21 strains belonged to other less prevalent serogroups. These strains belonged to diverse PhP/PCR types and did not show any similarity to the strains of two major serogroups, O114 and O127. The results suggest that two clonal groups of EPEC strains are predominantly associated with childhood diarrhoea in Bangladesh. PMID- 10670570 TI - Spread of the Brazilian epidemic clone of a multiresistant MRSA in two cities in Argentina. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is recognised as an important cause of nosocomial infection. The spread of some MRSA epidemic clones is well documented. In Brazil, and more recently in Portugal, a considerable number of hospital infections has been caused by a unique multiresistant MRSA clone designated as the Brazilian epidemic clone. This paper describes the spread of this clone in hospitals in two cities in Argentina. PMID- 10670571 TI - Distribution and expression of bca, the gene encoding the c alpha protein, by Streptococcus agalactiae. AB - A total of 52 clinical isolates of group B streptococci (GBS) was tested for expression of the c protein c(alpha) by a fluorescent antibody test (FAT) and by PCR amplification of a 202-bp stretch within the repeat unit of the bca gene encoding the c(alpha) protein. The strains were categorised as follows: c(alpha) FAT positive and PCR positive with amplification products of multiple sizes (category A, n = 12); FAT negative and with PCR products of multiple sizes (category B, n = 11); FAT negative and with a single PCR product of c. 200 bp (category C, n = 5); negative in both tests (category D, n = 24). A single amplification product of minimum size and additional products of larger sizes corresponded to one and more bca repeats, respectively. Five of the 11 category B strains showed expression of low Mr c(alpha) in whole cell-based Western blotting. The results showed that a proportion of the GBS isolates harboured bca gene elements that either were not expressed or they expressed c(alpha) molecular variants which could not be detected by the whole cell-based FAT. This genotype/phenotype discrepancy should be considered in relation to GBS typing, including the selection of antibody reagents and the technical approach to c(alpha) protein detection. PMID- 10670572 TI - Phenotypic characterisation of Candida albicans isolated from chronic hyperplastic candidosis. AB - The phenotypes of 35 Candida albicans isolates from 19 patients with chronic hyperplastic candidosis (CHC) and 35 isolates from 30 patients with non-CHC infections were compared. Typing was based on carbohydrate assimilation, chemical sensitivity and serology. Eight carbohydrate assimilation profiles were evident with the API-20C system and a single profile predominated for isolates from CHC (17 of 19 patients; 89%) and non-CHC (18 of 30 patients; 63%). Chemical sensitivity tests revealed four profiles with no significant difference between CHC and non-CHC isolates. Serotype A predominated for isolates from both CHC (15 of 19 patients; 79%) and non-CHC (25 of 30 patients; 83%) infections. Boric acid resistance was more prevalent in CHC isolates, although a significant difference was not apparent. In summary, there was no overall difference in the phenotypes of isolates from CHC and non-CHC patients, and clonal restriction of CHC isolates was not demonstrated. PMID- 10670573 TI - c-Kit and c-kit mutations in mastocytosis and other hematological diseases. AB - Mast cells (MC) are tissue elements derived from hematopoietic stem cells. Their differentiation and proliferation processes are under the influence of cytokines, including one of utmost importance known as stem cell factor (SCF). SCF receptor is encoded by the protooncogene c-kit, belongs to the type III receptor tyrosine kinase subfamily, and is also expressed on other hematopoietic or non hematopoietic cells. Ligation of c-kit receptor by SCF induces its dimerization, followed by induction of multiple intracellular signaling pathways leading to cell proliferation and activation. Mastocytosis, a relatively rare group of diseases characterized by accumulation of MC in various tissues, are found isolated or sometimes associated with other hematological malignancies in humans. Although the initial events leading to mastocytosis are not yet unraveled, alterations of the c-kit gene have been described. Particularly interesting are acquired mutations resulting in a constitutively activated receptor, possibly involved in the increased numbers of MC in tissues. For this reason, future strategies might be envisaged to target specifically the mutated c-kit and/or its intracellular signaling. PMID- 10670574 TI - Extracellular matrix moieties, cytokines, and enzymes: dynamic effects on immune cell behavior and inflammation. AB - Tissue injury caused by infection or physical damage evokes inflammatory reactions and events that are necessary for regaining homeostasis. Central to these events is the translocation of leukocytes, including monocytes, neutrophils, and T lymphocytes, from the vascular system, through endothelium, and into the extracellular matrix (ECM) surrounding the injured tissue. This transition from the vasculature into the site of inflammation elicits remarkable changes in leukocyte behavior as cells adhere to and migrate across ECM before carrying out their effector functions. Growing evidence suggests that, through its interactions with cytokines and degradative enzymes, the ECM microenvironment has a specialized role in providing intrinsic signals for coordinating leukocyte actions. Recent advances also reveal that enzymatic modifications to ECM moieties and cytokines induce distinctive cellular responses, and are likely part of the mechanism regulating the perpetuation or arrest of inflammation. This article reviews the findings that have elucidated the dynamic relationships among these factors and how they communicate with immune cells during inflammation. PMID- 10670575 TI - Reduced brain edema after traumatic brain injury in mice deficient in P-selectin and intercellular adhesion molecule-1. AB - Platelet (P-) selectin and intercellular adhesion molecule-1 (ICAM-1) mediate accumulation of neutrophils in brain. However, the mechanisms regulating neutrophil accumulation and damage after traumatic brain injury (TBI) are poorly defined. We hypothesized that mice deficient in both P-selectin and ICAM-1 (-/-) would have decreased brain neutrophil accumulation and edema, and improved functional and histopathological outcome after TBI compared with wild-type (+/+). In Protocol I, neutrophils and brain water content were quantified at 24 h after TBI. No difference in brain neutrophil accumulation was observed between groups; however, brain edema was decreased in dual P-selectin and ICAM-1 -/- (P < 0.05 vs. +/+ mice). In Protocol II, after TBI, tests of motor and memory function and histopathology were assessed over 21 days. No difference in motor or memory function or histopathological damage was observed between +/+ and -/- mice. A role for adhesion molecules in the pathogenesis of brain edema independent of leukocyte accumulation in brain is suggested. PMID- 10670576 TI - Homing and adhesion molecules in autoimmune gastritis. AB - The pathogenesis of autoimmune gastritis is the result of lymphocyte infiltration of the gastric mucosa, however, the events leading to the selective extravasation of autoreactive lymphocytes are unclear. Here we have examined the expression of adhesion molecules in the gastric mucosa of BALB/c mice with neonatal thymectomy induced gastritis. The overall area of vascular endothelium was not significantly different between gastritic and non-gastritic mice. However, a significant increase in the area of mucosal endothelium expressing MAdCAM-1 in gastritic mice was observed. Treatment of neonatally thymectomized BALB/c mice with a MAdCAM-1 specific monoclonal antibody (MECA 367) reduced the incidence of autoimmune gastritis from 80 to 26%. Treatment with a monoclonal antibody (R1-2) directed to the MAdCAM-1 ligand, alpha4beta7, also resulted in a reduction in the incidence of gastritis to 40%. These findings identify the alpha4beta7/MAdCAM-I interaction as a pivotal event in the initiation of autoimmune gastritis. PMID- 10670577 TI - In vivo administration of GM-CSF promotes the clearance of apoptotic cells: effects on monocytes and polymorphonuclear leukocytes. AB - The clearance of apoptotic cells is crucial to avoid chronic inflammation and autoimmunity. Little is known about the factors that regulate it in vivo. We show that granulocyte-macrophage colony-stimulating factor (GM-CSF) administration to carcinoma patients confers to their leukocytes a significantly higher ability to phagocytose apoptotic cells than before (P < 0.005). GM-CSF increased the concentration of monocytes and polymorphonuclear leukocytes in the peripheral blood and activated circulating polymorphonuclear leukocytes. Both effects abated early after treatment, whereas phagocytosis of apoptotic cells was still significantly higher after 18 days compared with basal values (P < 0.005 and P < 0.025 for monocytes and polymorphonuclear leukocytes, respectively). On in vitro phagocytosis of apoptotic cells monocytes, but not polymorphonuclear leukocytes, up-regulated MHC class II membrane expression. These findings are consistent with the possibility that GM-CSF endows both scavenger and antigen-presenting leukocytes with the ability to internalize apoptotic tumor cells. PMID- 10670578 TI - Serglycin secreted by leukocytes is efficiently eliminated from the circulation by sinusoidal scavenger endothelial cells in the liver. AB - This study was undertaken to determine the fate of the circulating chondroitin sulfate proteoglycan serglycin. The human monocytic cell line THP-1 was cultured under serum-free conditions in the presence of [35S]sulfate. The conditioned medium was harvested and 35S-macromolecules were purified by Q-Sepharose anion exchange chromatography and Superose 6 gel chromatography. After labeling with 125I, the purified material was treated with chondroitinase ABC and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. A major band with mr of approximately 14 kDa appeared, consistent with the core protein of serglycin. The identity of the proteoglycan was confirmed by amino-terminal amino acid sequencing. Purified serglycin, labeled either with [35S]sulfate or 125I and fluorescein isothiocyanate, was injected intravenously into rats. The blood content of radiolabeled serglycin fell by 50% from 1 to 2.4 min after injection, indicating an initial t1/2 of 1.4 min or shorter. Approximately 90% of the recovered radioactivity was localized in the liver, 5% in the blood, and 5% altogether in urine, kidneys, and spleen about 30 min after injection. Isolation of liver cells at the same time point showed that 70% of the radioactivity was taken up by the sinusoidal scavenger endothelial cells, and 23 and 7% by the hepatocytes and Kupffer cells, respectively. When excess amounts of unlabeled hyaluronan was coinjected with radiolabeled serglycin, the elimination of serglycin was significantly inhibited, indicating that the hyaluronan receptor on the sinusoidal scavenger endothelial cells is responsible for the elimination of serglycin. PMID- 10670579 TI - Role of endothelins on lymphocyte accumulation in allergic pleurisy. AB - Endothelins participate in different aspects of inflammatory reactions, including edema formation and eosinophil accumulation in allergic reaction. In this study, we demonstrated a role for endogenous endothelins in eosinophil and T lymphocyte recruitment and cytokine secretion in a murine model of allergic inflammation. Intrathoracic stimulation with endothelin-1 triggered a neutrophil accumulation at 4 h, concomitant with an increase of CD4+ and CD8+ T lymphocyte populations. Antigen challenge in sensitized animals leads to an increase in eosinophil and mononuclear cell numbers at 24 h. Treatment with ETA receptor antagonist (BQ123) inhibited antigen-induced eosinophil and mononuclear cell migration, whereas the selective ETB receptor antagonist BQ-788 was ineffective. The latter effect of BQ 123 was due to inhibition of CD4+ and CD8+ T lymphocytes. Treatment with BQ-123 also inhibited interleukin-5 levels in the exudate and plasma as well as intracellular staining of interleukin-4, interleukin-5, and interferon-gamma in CD4+ lymphocytes. These findings suggest that endogenous endothelins contribute to allergic inflammation by modulating lymphocyte recruitment and cytokine production. PMID- 10670580 TI - CD11b/CD18-coated microspheres attach to E-selectin under flow. AB - Neutrophils can attach to E-selectin under flow. Proposed ligands for E-selectin carry SLe(x)-type glycans. The leukocyte beta2 integrins are glycosylated with SLe(x). Thus, we speculated that beta2 integrins could support attachment to E selectin. To test this hypothesis, we coated 10-microm-diameter microspheres with purified CD11b/CD18 (alphaMbeta2) and investigated the adhesion of the resulting alphaMbeta2 microspheres to E-selectin. Under in vitro flow conditions, the alphaMbeta2 microspheres attached to Chinese hamster ovary cells expressing E selectin (CHO-E) and 4-h interleukin-1beta-activated human umbilical vein endothelial cells (HUVEC). At a shear stress of 1.8 dynes/cm2, the attachment events were eliminated by pretreatment of the cellular monolayers with a mAb to E selectin. alphaMbeta2 microspheres did not attach to untransfected CHO cells or unactivated HUVEC at 1.8 dynes/cm2. Taken together, the results strongly suggest that the CD11b/CD18-E-selectin bond has sufficient biophysical properties to mediate attachment of neutrophil-sized particles to E-selectin under flow. PMID- 10670581 TI - Langerhans cells acquire a CD8+ dendritic cell phenotype on maturation by CD40 ligation. AB - Dendritic cell (DC) reconstitution experiments and phenotypic analysis of DC subpopulations have allowed the definition in the mouse of two main DC categories: CD8+ lymphoid DCs and CD8- myeloid DCs. With regard to Langerhans cells (LCs), which represent immature DCs differentiating into mature DCs on migration to the lymph nodes after an antigenic stimulation, although classically considered as myeloid DCs, there is no experimental evidence of their origin. It has been recently shown that mouse LCs, negative for CD8 and LFA-1, undergo CD8/LFA-1 up-regulation on migration, suggesting that LCs belong to the CD8+ lymphoid DC lineage. To further reinforce this hypothesis, we have analyzed the modulation of CD8 expression by LCs on culture with molecules known to induce LC maturation. Our results show that LC acquired a CD8+ lymphoid phenotype on CD40 ligation. PMID- 10670582 TI - P47(phox)-deficient NADPH oxidase defect in neutrophils of diabetic mouse strains, C57BL/6J-m db/db and db/+. AB - Deficiencies in neutrophil NADPH oxidase proteins have been demonstrated in humans with chronic granulomatous disease. However, no spontaneous mutation in murine NADPH oxidase has been reported. In this study we report that neutrophils from the diabetic mouse strains, C57BL/6J-m heterozygous lean (lepr(db/+)) and homozygous obese (lepr(db/db)) mice produced no superoxide on stimulation. An absence of intact p47(phox) but not other oxidase proteins was observed in both mouse strains through the use of immunoblotting. Molecular analysis by reverse transcriptase-polymerase chain reaction identified three abnormal p47phox mRNA transcripts. Sequencing of genomic DNA of p47(phox) revealed a point mutation at the -2 position of exon 8, which is consistent with aberrant splicing of the p47(phox) transcript. These results indicate that the C57BL/6J-m db/db and db/+ mice are the first spontaneously derived murine model of NADPH oxidase deficiency involving a p47(phox) mutation. PMID- 10670583 TI - Differential role of tyrosine phosphorylation in adhesion-induced transcription, mRNA stability, and cytoskeletal organization in human monocytes. AB - Monocyte adhesion resulted in rapid tyrosine phosphorylation and subsequent cytokine mRNA induction. The objective of this study was to determine the role of specific tyrosine phosphorylation events, particularly those involving members of the MAP kinase family, in regulating adhesion-induced cytokine expression. Using nuclear run-on analyses, we demonstrated that on adhesion, monocytes rapidly transcriptionally activated numerous cytokine mRNAs, coincident with the activation of the transcription factors NF-KB and AP-1. Both an inhibitor of tyrosine phosphorylation, genistein, and the cytoplasmic tyrosine phosphatase PTP1B, were unable to prevent adhesion-mediated transcriptional activation. However, both blocked adhesion-induced ERK and JNK but not p38 kinase activation and at the same time decreased the stability of interleukin-1beta (IL-1beta) and IL-8 transcripts. In addition, whereas adhesive events occurred in the presence of genistein and PTP1B, monocyte spreading was markedly inhibited. Our results suggest that the majority of protein phosphorylation events are associated with adhesion-induced cytokine expression through transcript stabilization and cytoskeletal organization. A minority of protein phosphorylation events, not sensitive to genistein or PTP1B exposure, may be instrumental in regulating transcription. Thus the spectrum of protein tyrosine kinases required for transcription appear distinct from those involved in maintaining the stability of some cytokine mRNAs and the integrity of the cytoskeleton to which mRNA destined for translation must be associated. PMID- 10670584 TI - Particulate adjuvants can induce macrophage survival, DNA synthesis, and a synergistic proliferative response to GM-CSF and CSF-1. AB - The mode of action of immunological adjuvants is not yet completely understood. Many are particulate. Certain antigen-presenting (dendritic) cell populations belong to the monocyte/macrophage lineage and, like other members of the lineage, in some tissues appear to be short-lived. We report that many poorly degradable, particulate adjuvants, for example, aluminum hydroxide, oil-in-water emulsions, calcium phosphate, and silica, enhance murine bone marrow-derived macrophage survival; induction of DNA synthesis was even observed. No evidence could be found for a requirement for endogenous granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage-CSF (M-CSF or CSF-1). Synergy for the proliferative effects was noted in the presence of added GM-CSF or CSF-1. It is suggested from these in vitro findings that one function of certain particulate adjuvants may be to increase by enhanced survival or even proliferation the number of cells available for subsequent antigen presentation and cytokine production. PMID- 10670585 TI - Antimicrobial effects of alpha-MSH peptides. AB - The presence of the ancient anti-inflammatory peptide alpha-melanocyte stimulating hormone [alpha-MSH (1-13), SYSMEHFRWGKPV] in barrier organs such as gut and skin suggests a role in the nonspecific (innate) host defense. alpha-MSH and and its carboxy-terminal tripeptide (11-13, KPV) were determined to have antimicrobial influences against two major and representative pathogens: Staphylococcus aureus and Candida albicans. alpha-MSH peptides significantly inhibited S. aureus colony formation and reversed the enhancing effect of urokinase on colony formation. Antimicrobial effects occurred over a broad range of concentrations including the physiological (picomolar) range. Small concentrations of alpha-MSH peptides likewise reduced viability and germ tube formation of the yeast C. albicans. Antimicrobial influences of alpha-MSH peptides could be mediated by their capacity to increase cellular cAMP. Indeed, this messenger was significantly augmented in peptide-treated yeast and the potent adenylyl cyclase inhibitor dideoxyadenosine (ddAdo) partly reversed the killing activity of alpha-MSH peptides. Reduced killing of pathogens is a detrimental consequence of therapy with anti-inflammatory drugs. Because alpha MSH has potent anti-inflammatory effects we determined influences of alpha-MSH on C. albicans and S. aureus killing by human neutrophils. alpha-MSH peptides did not reduce killing but rather enhanced it, likely as a consequence of the direct antimicrobial activity. alpha-MSH peptides that combine antipyretic, anti inflammatory, and antimicrobial effects could be useful in treatment of disorders in which infection and inflammation coexist. PMID- 10670586 TI - Expression of serum- and glucocorticoid-regulated kinase (sgk) mRNA is up regulated by GM-CSF and other proinflammatory mediators in human granulocytes. AB - Stimulation of human peripheral blood granulocytes with the proinflammatory cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF), increases incorporation of [3H]uridine into RNA. We investigated the nature of the RNA synthesized under these conditions. Using transcription inhibitors, gel electrophoresis, and high-salt precipitation, it was concluded that as much as 90% of this radiolabeled RNA represents polymerase II transcripts. Differential display reverse transcription-polymerase chain reaction was used to identify and clone GM-CSF-responsive mRNAs. Serum- and glucocorticoid-regulated kinase (sgk) mRNA was identified that could be up-regulated 10- to 20-fold by > or =0. 1 ng/mL recombinant human GM-CSF. The 2.6-kb sgk mRNA was induced rapidly (within 30 min) by GM-CSF and remained at high levels for at least 12 h. Up-regulation was blocked completely by the transcription inhibitor, actinomycin D, but not by the translation inhibitor, cycloheximide, nor by the tyrosine kinase inhibitor, genistein. Up-regulation did not appear to be caused by enhanced mRNA stability. Other inflammatory mediators could also increase sgk mRNA levels (GM-CSF > > lipopolysaccharide > fMLP = tumor necrosis factor alpha). The function of sgk in granulocytes remains unknown. PMID- 10670587 TI - Rottlerin, a PKC isozyme-selective inhibitor, affects signaling events and cytokine production in human monocytes. AB - The implication of select protein kinase C (PKC) isoenzymes in cytokine production by human monocytes was investigated using an isozyme-selective inhibitor of PKC, rottlerin. We found that lipopolysaccharide (LPS) triggers cytosol-to-membrane translocation of PKCalpha and delta isoenzymes, whereas phorbol ester (PMA) induces translocation of several PKC isoforms. Moreover, we show that in LPS- and PMA-stimulated monocytes rottlerin affects several cellular responses. (1) At low (15 microM) concentration it blocks translocation of PKCdelta, diminishes DNA binding activity of AP-1 transcription factor, and attenuates cytokine production [tumor necrosis factor alpha (TNF-alpha) > interleukin-1beta (IL-1beta)]. (2) At high (50 microM) concentration it prevents translocation of PKCalpha, and subsequently inhibits ERK1/ERK2 phosphorylation, DNA binding activities of AP-1 and nuclear factor-KB transcription factors, and the production of both tested cytokines. Thus, we propose that cytosol-to membrane translocation of PKCalpha and PKdelta isoenzymes may represent early steps in the signaling cascades that lead to TNF-alpha and IL-1beta production in human monocytes. PMID- 10670589 TI - Facing the problems of faculty recruitment and retention. PMID- 10670588 TI - AP-1 activity is negatively regulated by cannabinol through inhibition of its protein components, c-fos and c-jun. AB - Regulation of the activator protein-1 (AP-1) complex is very intricate because it involves phosphorylation state, protein-protein, and protein-DNA interactions. In these studies, the regulation of AP-1 activity, with emphasis on c-fos and c-jun regulation, was investigated using cannabinol (CBN) in primary mouse splenocytes in vitro. Cannabinoid compounds exhibit immunosuppressive actions that are putatively mediated through Gi-protein coupled receptors that negatively regulate adenylate cyclase. However, recent studies suggest that cannabinoids modulate other signaling cascades. Indeed, we demonstrate that CBN inhibited binding to AP 1-containing sites from the interleukin-2 promoter. This inhibition of binding was, in part, due to decreased nuclear expression of c-fos and c-jun. We further determined that the effects of CBN were due to posttranslational modifications of these phosphoproteins and showed that CBN inhibited the activation of ERK MAP kinases. Thus, cannabinoid-induced immunosuppression involves disruption of the ERK signaling cascade. PMID- 10670590 TI - Anticoagulation and minor oral surgery: should the anticoagulation regimen be altered? AB - PURPOSE: This study was performed to assess the extent of bleeding in anticoagulated patients undergoing minor oral surgery procedures when compared with patients who stop their anticoagulation regimen before surgery and patients who have never been anticoagulated. MATERIALS AND METHODS: Patients on anticoagulant medications undergoing dentoalveolar surgery procedures either stopped their anticoagulation regimen 72 to 96 hours before the planned surgical intervention or continued their regular regimen throughout the time of surgery. Blood loss was measured by weighing sponges used in the procedures, and groups were compared for differences in blood loss. They were also compared with an additional control group that had never been on anticoagulant therapy. RESULTS: There was no difference in blood loss detected among any of the experimental or control groups. No bleeding complications occurred in any anticoagulated patient. CONCLUSIONS: The data suggest that many patients can safely undergo routine outpatient oral surgical procedures without alteration of their regular therapeutic anticoagulation regimens and without additional medical intervention. However, a larger experimental population may be needed to elucidate the appropriateness of this approach to perioperative care. PMID- 10670591 TI - Obesity: prevalence and considerations in oral and maxillofacial surgery. AB - PURPOSE: This article examines the prevalence of obesity in an urban hospital based office population and describes the associated health risks and systemic complications. PATIENTS AND METHODS: A total of 225 randomly selected patient charts were reviewed. The patients' height and weight were recorded, and a body mass index (BMI) was calculated. Patients were considered to have a normal weight with a BMI between 20 and 24.9 kg/m2, to be overweight with a BMI between 25 and 29.9 kg/m2, and to be obese with a BMI of 30 kg/m2 or greater. Comparisons between sex, age, and race were examined. RESULTS: Eighty-five males and 140 females were included in the study. The ages ranged from 9 to 86 years, with a mean age of 37.4+/-16.4 years. The BMIs ranged from 13.9 to 57.7 kg/m2, with a mean BMI of 26.5+/-6.8 kg/m2. Over half (51%) of the population studied was overweight, and 23% were obese. Forty-three percent of males were considered overweight, and 18.8% of males were obese. Women tended to be more overweight: 55.7% of women were overweight and 25.7% were obese. The African-American females had mean BMIs that were considered overweight in all age-groups and obese in most age-groups, which included the less than 29-year-old and over 50-year-old groups. Caucasian females had normal mean BMIs at all ages except for the 30- to 39-year old and 40- to 49-year-old groups. CONCLUSION: The increasing prevalence of overweight and obese populations has several considerations in oral and maxillofacial surgery. The associated health risks and increased morbidity and mortality pose a serious threat to the patient being treated in an outpatient setting. PMID- 10670592 TI - Cervical necrotizing fasciitis of odontogenic origin: a case report and review of 12 cases. AB - PURPOSE: This article reviews the demographics, presentation, cause, clinical findings, and treatment of 12 cases of cervical necrotizing fasciitis of odontogenic origin. PATIENTS AND METHODS: A retrospective chart review of 12 cases treated between 1987 and 1997 was done. RESULTS: Most cases resulted from an abscessed mandibular molar. The most common significant medical conditions in the patient's history were diabetes, hypertension, obesity, and substance abuse. All patients were treated surgically within 24 hours of admission. Hyperbaric oxygen (HBO) was used as adjunctive treatment in all cases. The average length of hospital stay was 31 days. All patients recovered. CONCLUSION: Early surgical intervention and the use of HBO decreases morbidity and improves the clinical outcome. PMID- 10670593 TI - Nutritional status of substance abusers with mandible fractures. AB - PURPOSE: The purposes of this study were 1) to assess the validity of patient self-report in identifying illegal substance abuse and 2) to identify nutritional deficiencies in substance abusers presenting for treatment of mandible fractures. PATIENTS AND METHODS: To address the research purposes, a prospective cohort study was conducted of patients presenting for treatment of mandible fractures. A urine drug screen was used to determine the validity of patient self-report of substance abuse. For purposes of assessing nutritional status, 2 categories of substance abusers were identified: illegal and legal (alcohol). The nutritional status was measured using various laboratory markers. RESULTS: The sample was composed of 93 subjects. Urine drug studies were available for 32 patients. Of the 22 patients who denied illegal drug use, 12 (55%) had a positive drug screen. Of the 10 patients reporting a positive history of illicit drug use, 7 (70%) had a positive urine drug screen (P = .47). A positive correlation was found between alcohol exposure and serum aspartate aminotransferase, mean corpuscular volume, and lactate dehydrogenase. Positive drug screens also were associated with increased serum ferritin levels. CONCLUSIONS: The results of this study suggest that patient self-report of illicit drug use may be unreliable. The findings also suggest that legal and illegal substance abusers presenting for treatment of mandible fractures have minimal nutritional deficiencies. PMID- 10670594 TI - The use of the buccal fat pad for reconstruction of oral defects: review of the literature and report of 15 cases. AB - PURPOSE: Although the buccal fat pad (BFP) was originally used as an alternative method for the closure of small to medium-sized oroantral and oronasal communications, its use has now been extended to use after excision of oral malignancies. This report describes experience with this technique. PATIENTS AND METHODS: The BFP was used as a pedicled graft to reconstruct medium-sized surgical defects of the oral soft and hard tissues in 15 patients suffering from oral malignant tumors. Six of the defects were in the maxilla, 3 in the retromandibular area, and 6 in the cheek and oral commissure. The BFP was left uncovered to epithelialize in 14 patients, and in one it was covered with lyophilized dura. RESULTS: The BFP healed without complications within 3 to 4 weeks in 13 patients, whereas in 2 patients with maxillary defects there was partial loss of the graft, requiring the additional use of an obturator in 1 case and a tongue flap in another to prevent oronasal leakage. Harvesting the graft proved to be extremely easy, and care was only necessary to avoid severing the supporting vascular plexus and the thin capsule covering the BFP. CONCLUSIONS: The findings support the view that the BFP is a useful, easy, and uncomplicated alternative method for the reconstruction of small to medium-sized surgical defects of the oral hard and soft tissues. PMID- 10670595 TI - Free fat transplantation for facial tissue augmentation. AB - PURPOSE: This article reviews the lipofilling technique using a fat tissue suspension obtained by manual or vacuum machine-assisted suction. PATIENTS AND METHODS: Between 1989 and 1997 14 patients were treated by lipofilling. All patients were female; 6 were treated for cheek and zygomatic area augmentation for Romberg syndrome (2 cases) and bilateral facial lipodystrophy (4 cases); 4 were treated for nasolabial fold correction, 3 for lip augmentation, and 1 for correction of systemic lupus erythematosus (LES) scars. RESULTS: Good results were obtained in the treatment of the moderate Romberg syndrome case, with 3 stages of fat tissue transplantation from the abdominal area, in the severe Romberg syndrome case that was treated with 3 stages of lipofilling after a primary surgical procedure consisting of augmentation of the zygomatic region using a composite temporal flap of calvaria bone and temporal muscle, in the 4 cases of bilateral facial lipodystrophy, and in the 4 cases of nasolabial fold correction with 3 to 4 stages of lipofilling. Poor results were obtained in lip augmentation, and bad results were obtained in correction of the LES scars. CONCLUSIONS: Using the lipofilling technique with cannulas of 3 to 4 mm in diameter and low-power manual aspiration to preserve the integrity of the fat cells, good results were achieved in 10 of the 14 patients treated. The poor results occurred in areas of reduced vascularity and high motility. PMID- 10670596 TI - Force level and strain patterns during bilateral mandibular osteodistraction. AB - PURPOSE: Recent reports have demonstrated that device orientation is important during mandibular distraction osteogenesis. The purpose of this study was to evaluate the force level and strain patterns on the mandible during bilateral osteodistraction with devices oriented either parallel to the body of the mandible or parallel to the sagittal axis of distraction. MATERIALS AND METHODS: Five unembalmed human cadaver mandibles were placed in a specially designed apparatus for stabilization of the proximal segments during distraction. A force transducer was attached to the lateral aspect of the inferior ramus, and strain gauges were attached to the mandibular bone segments proximal and distal to the distraction device. Lateral force and bone strains were then measured at 5 and 10 mm of distraction. Osteodistraction proceeded first with the devices placed parallel to the mandibular body, then parallel to the axis of distraction. RESULTS: Significantly greater lateral forces were seen when the devices were oriented parallel to the mandibular body. With this device orientation, increased tensile strains were seen at the labial symphysis and medial ramus, and increased compressive strains were found at the lingual symphysis and lateral ramus. However, when the devices were oriented parallel to the axis of distraction, the forces and strains were not detected. CONCLUSIONS: The results suggest that device orientation has important biomechanical effects on lateral forces and strain patterns during mandibular osteodistraction. PMID- 10670597 TI - Prostaglandin E2 in temporomandibular joint synovial fluid and its relation to pain and inflammatory disorders. AB - PURPOSE: The aim of this study was to investigate temporomandibular joint (TMJ) synovial fluid (SF) levels of prostaglandin E2 and its relation to general inflammatory activity and its influence on specific TMJ pain in patients with inflammatory TMJ disorders. PATIENTS AND METHODS: The study comprised 24 patients (30 joints) with inflammatory TMJ disorders and 4 healthy persons (6 joints). TMJ pain at rest, tenderness to palpation of the TMJ, and TMJ pressure pain threshold, as well as pain during joint movements (PM), were assessed. PGE2 levels were analyzed in synovial fluid samples (SF-PGE2) and blood plasma (P PGE2). The erythrocyte sedimentation rate (B-ESR) as well as the serum levels of C-reactive protein (S-CRP) and antinuclear antibodies were determined. RESULTS: PGE2 was undetectable in the plasma and in the TMJ SF of the healthy persons. In the patients, PGE2 was detectable in 20 of the 30 (67%) SF samples. SF-PGE2 was significantly and positively correlated to PM in the patients. There were significant correlations between P-PGE2 and B-ESR as well as the S-CRP. CONCLUSIONS: This study shows that the synovial fluid in patients with TMJ inflammatory disorders frequently has a detectable level of PGE2 that is related to TMJ allodynia. The plasma levels of PGE2 seem to be related to the general inflammatory activity in these patients. PMID- 10670598 TI - Effect of estrogen replacement on temporomandibular joint remodeling in ovariectomized rats. AB - PURPOSE: The investigation was performed to elucidate the effect of estrogen on the temporomandibular joint (TMJ) and to evaluate the therapeutic effect of 17beta-estradiol replacement in growing rats. MATERIALS AND METHODS: Thirty 4 week-old female albino Wistar rats were divided into 3 groups. Ten rats were ovariectomized followed by intramuscular administration of 17beta-estradiol for hormone replacement (OVX + E2), 10 were sham operated (CTL), and 10 were ovariectomized without hormone replacement (OVX). Five rats from each group were killed at 1 and 2 weeks postoperatively, and the serum estrogen was determined to verify the adequacy of replacement. The temporomandibular joints of the age matched sham-operated control and ovariectomized groups were histomorphometrically evaluated at the same periods. RESULTS: In OVX animals, the thickness of the articular soft tissue was increased by a concomitant increase of the transitional and cartilage zones in the anterior and posterior portions at 1 and 2 weeks postoperatively. However, the bone volume was decreased in the anterior and posterior portions at 2 weeks after the surgery and the condyle was flattened. Replacement with 17beta-estradiol restored most of the histomorphometric parameters. The thickness of articular soft tissue was increased in the anterior portion by an increase in the cartilage zone in the OVX + E2 group at 2 weeks postoperatively. Increase of bone volume was found at 2 weeks after hormone replacement with a corresponding increased osteoid surface and decreased quiescent surface in the central portion at 1 week postoperatively. A flattened condyle was still noted at 2 weeks postoperatively in the OVX + E2 animals despite the hormone replacement. CONCLUSIONS: Estrogen in a physiologic concentration may play an important role in TMJ remodeling. Progesterone may be indispensable for remodeling, particularly contributing to morphogenesis. PMID- 10670599 TI - Management of heparin therapy in the high-risk, chronically anticoagulated, oral surgery patient: a review and a proposed nomogram. AB - PURPOSE: This study analyzes the use of a standard nomogram that can help reduce the level of anticoagulation preoperatively to effectively manage perioperative heparin therapy in chronically anticoagulated oral surgery patients who are at high risk for thromboembolism. PATIENTS AND METHODS: Twenty patients with significant cardiovascular disease, ranging in age from 56 to 79 years and requiring oral surgery, were randomly divided into 2 groups. All patients were on chronic warfarin therapy, and perioperative heparinization was recommended by their cardiologist. Group A (n = 10) had their anticoagulation therapy managed with the use of a standard nomogram. The heparin therapy for group B (n = 10) was managed without the use of the nomogram. The records of all patients were analyzed for therapeutic efficacy of heparinization, number of laboratory tests required, duration of hospitalization, and complications related to heparinization. RESULTS: Patients in group A did significantly better in all parameters when compared with group B patients. There were no complications in group A, whereas there was a 20% incidence of complications related to anticoagulation therapy in group B. CONCLUSIONS: The use of a standard nomogram to manage anticoagulation therapy in the oral surgery patient requiring heparinization is strongly recommended. This provides optimal therapeutic benefit, decreases the incidence of complications, and makes the hospitalization less costly and more comfortable for the patient. PMID- 10670600 TI - Tender swelling of the chin 40 years after genioplasty. PMID- 10670601 TI - Current trends in the treatment of maxillofacial injuries in the United States. AB - This report presents the results of a 1997 survey of the members of the America Association of Oral and Maxillofacial Surgeons to assess the current national and regional trends in the management of maxillofacial trauma. Comparisons are made with the nearly identical survey done in 1987. The results show practitioner age related differences and changes in reported practice across time. The significance of these findings are discussed. PMID- 10670602 TI - Ganglion cyst of the temporomandibular joint: report of case and review of literature. PMID- 10670603 TI - Management of early relapse after a sagittal split ramus osteotomy by gradual callus distraction: a case report. PMID- 10670604 TI - Surgical management of lingual thyroid: a report of four cases. PMID- 10670605 TI - Joint formation between an osteochondroma of the coronoid process and the zygomatic arch (Jacob disease): report of case and review of literature. PMID- 10670606 TI - Brown tumor of the maxilla in a patient with secondary hyperparathyroidism: a case study involving immunohistochemistry and electron microscopy. PMID- 10670607 TI - Traumatic intracranial impaction of the zygoma: case report. PMID- 10670608 TI - Descending necrotizing mediastinitis caused by an odontogenic infection: a case report. PMID- 10670609 TI - Is trabecular bone in the mandible different? PMID- 10670610 TI - Pesticide safety training. PMID- 10670611 TI - Survivors of torture. PMID- 10670612 TI - Involving a community in a marine safety investigation. PMID- 10670613 TI - 44.3 million in US lack health insurance. PMID- 10670615 TI - NIH launches new publishing website. PMID- 10670614 TI - GAO looks at state Medicaid enrollment post-welfare reform. PMID- 10670616 TI - Providers, health plans clash over patient care. PMID- 10670617 TI - Infertility: from a personal to a public health problem. AB - The inability to conceive a child is most often viewed as a private matter, but public health perspectives and skills can contribute greatly to our knowledge about infertility, and the development of effective and rational public policy for prevention, access to health care, and regulation of new technologies. We offer a primer of public health aspects of infertility in an effort to encourage the broad spectrum of public health professionals to become more knowledgeable about these topics and join in the national debate about preventive strategies, cost-benefit assessment, resource allocation, and ethics. PMID- 10670618 TI - Mercury risks: controversy or just uncertainty? PMID- 10670619 TI - Winston's "No Additives" campaign: "straight up"? "no bull"? AB - OBJECTIVE: The author used data from a larger study to examine adolescents' and adults' responses to Winston cigarettes' "No Additives" advertising campaign. METHODS: The author analyzed responses from 400 adolescents ages 12-17 and 203 adults ages 30-50 who were asked what they believed the meaning of the "No Additives" slogan to be. The author also analyzed adolescents' responses to questions about four specific Winston "No Additives" ads. RESULTS: Two-thirds of adolescents and 27% of adults believed that "No Additives" meant one or more of the following: that Winston cigarettes are healthier than other cigarettes, that they are less likely to harm health, or that they are less likely to be addictive. Adolescents perceived the models in three ads to be younger than 25 years old. Among adolescent respondents, smokers were more likely than nonsmokers to like the ads and to believe the ads made smoking more appealing. CONCLUSIONS: The "No Additives" slogan was perceived by a majority of adolescents and about a quarter of adults as implying one or more health claims. The results of this analysis suggest that the Federal Trade Commission's action in requiring a disclaimer on the "No Additives" ads is well founded but the disclaimer should be strengthened. PMID- 10670620 TI - A community strategy for Medicaid child dental services. AB - OBJECTIVES: The authors present second-year utilization data and first- and second-year cost data for a community-based program in Spokane County, Washington, designed to increase access to dental care for Medicaid-enrolled children from birth to 60 months of age. METHODS: The authors used Medicaid eligibility and claims data for 18,727 children 5 years of age and younger to determine utilization of dental care from January 15, 1996, through January 15, 1997. They also used accounting records from the agencies involved to calculate the first- and second-year costs of the program. RESULTS: A child in the ABCD program was 7.2 times as likely to have at least one dental visit as a Medicaid enrolled child not in the program. Estimated costs per child with at least one dental visit (in 1995 dollars) were $54.30 for the first year and $44.38 for the second year, or $20.09 per enrolled child for the first year and $18.77 for the second year. CONCLUSION: Public-private joint efforts are effective in improving access to dental care for Medicaid-enrolled children. PMID- 10670621 TI - Reducing firearm injuries: the role of local public health departments. AB - OBJECTIVE: The purpose of this study was to gather data regarding local public health departments' involvement in activities to prevent firearm-related morbidity and mortality. METHODS: A questionnaire was sent to local public health departments serving cities with populations > or =60,000 to assess their perceptions of the magnitude of the firearm injury problem in their jurisdictions and the activities in which they were engaged to reduce firearm-related injuries. RESULTS: Almost half (49.7%) of respondents said that their departments had not seriously thought about being involved in activities to reduce firearm-related injuries, and fewer than one in five (17.8%) reported that their departments were involved in such activities. Respondents identified three barriers to involvement in activities to reduce firearm injuries: limited financial resources (62.7% of respondents), lack of expertise (50.8%), and not enough time (47%). CONCLUSIONS: Despite the extent of firearm injuries in the US, systematic collection of local data on firearm morbidity and mortality to help guide policy development is lacking. PMID- 10670622 TI - Welfare reform: advocacy and intervention in the health care setting. AB - Welfare reform has drastically altered the lives of poor families in the US. In its wake, many former recipients are not receiving whatever transitional benefits and other safeguards to which they remain entitled under federal and state laws. Families are losing access to Medicaid and are not receiving the child care assistance or Food Stamps for which they continue to be eligible. Ill-served by stringent time limits and work requirements, lack of child care assistance, and lack of training and educational opportunities for the development of skills that will lead to better jobs, families need help to navigate the complexities of the new welfare system. Boston Medical Center's Department of Pediatrics has instituted a welfare screening project to educate families about their rights under welfare reform and assist them in advocating for themselves and their children. PMID- 10670623 TI - Alaska's model program for surveillance and prevention of occupational injury deaths. AB - The National Institute for Occupational Safety and Health (NIOSH) established its Alaska Field Station in Anchorage in 1991 after identifying Alaska as the highest risk state for traumatic worker fatalities. Since then, the Field Station, working in collaboration with other agencies, organizations, and individuals, has established a program for occupational injury surveillance in Alaska and formed interagency working groups to address the risk factors leading to occupational death and injury in the state. Collaborative efforts have contributed to reducing crash rates and mortality in Alaska's rapidly expanding helicopter logging industry and have played an important supportive role in the substantial progress made in reducing the mortality rate in Alaska's commercial fishing industry (historically Alaska's and America's most dangerous industry). Alaska experienced a 46% overall decline in work-related acute traumatic injury deaths from 1991 to 1998, a 64% decline in commercial fishing deaths, and a very sharp decline in helicopter logging-related deaths. Extending this regional approach to other parts of the country and applying these strategies to the entire spectrum of occupational injury and disease hazards could have a broad effect on reducing occupational injuries. PMID- 10670624 TI - A forgotten enemy: PHS's fight against the 1918 influenza pandemic. PMID- 10670625 TI - A collaborative effort to develop a data collection system. PMID- 10670627 TI - Transplantation of highly differentiated immortalized human hepatocytes to treat acute liver failure. AB - BACKGROUND: Temporary support of a damaged liver by a bioartificial liver (BAL) devise is a promising approach for the treatment of acute liver failure. Although human primary hepatocytes are an ideal source of hepatic function in BAL, shortage of human livers available for hepatocyte isolation is the limiting factor for the use of this modality. A clonal human hepatocyte cell line that can grow economically in culture and exhibit liver-specific functions should be an attractive solution to this problem. METHODS: To test this alternative, primary human fetal hepatocytes were immortalized using Simian virus 40 large T antigen. To investigate the potential of the immortalized cells for BAL, we transplanted the cells into the spleen of adult rats and performed a 90% hepatectomy 12 hr later. RESULTS: One of the cloned human liver cell lines, OUMS-29, showed highly differentiated liver functions. Intrasplenic transplanting of 20x10(6) OUMS-29 cells protected the animals from hyperammonemia and the associated hepatic encephalopathy. Survival was significantly prolonged in 90% of hepatectomized rats receiving OUMS-29 cells. CONCLUSIONS: A highly differentiated immortalized human hepatocyte cell line, OUMS-29, was able to provide metabolic support during acute liver failure induced by 90% hepatectomy in rats. Essentially unlimited availability of OUMS-29 cells may be clinically useful for BAL treatment. PMID- 10670626 TI - Long-term functional islet mass and metabolic function after xenoislet transplantation in primates. AB - BACKGROUND: Pancreatic islet transplantation (PIT) is an attractive alternative for patients with type I diabetes mellitus. PIT is not yet an effective clinical reality due in part to the high incidence of rejection and early loss of functional islet mass. In addition, current immunosuppressive drugs have toxic effects on islets and increase the risk of morbidity and mortality. In the present study, the effects of PIT on glycemic parameters were assessed in spontaneously diabetic primates. METHODS: Five insulinopenic nonhuman primates (three Macacca fascicularis, one Ceropithecus aethiops, and one Macacca mulatta) were studied. All required twice-daily treatment with 4-10 U of insulin. For immunosuppression, the animals received anti-CD3-immunotoxin (100 microg/kg(initially infused 2 hr before transplantation and again on day +1), cyclosporine (CsA) (20 mg/kg(i.v./2 hr before transplantation), cyclosporine microemulsion (Neoral) 60 mg/kg/b.i.d. on days +1 to +3 with dose adjusted by blood levels, and methylprednisolone (15 mg/kg day 0 to +3). Three recipients were given islets from a single donor (M mulatta). The islets were prepared by a semiautomated technique using Liberase. A mean of 13,136 islet equivalents/kg was infused into the portal vein. Two animals (M fascicularis and M mulatta) were used as a diabetic, nontransplanted control. Several metabolic parameters were evaluated. RESULTS: All monkeys that underwent transplantation experienced reversal of diabetes mellitus with normalization of all diabetic glycemic parameters. In the nontransplanted primates given the same immunosuppression but no PIT, diabetic metabolic parameters were unchanged after 9 months of follow-up. In contrast, all three PIT recipients established fasting and nonfasting euglycemia within 1-2 weeks, and none required exogenous insulin after day 10. Normal intravenous glucose tolerance tests were observed at day 15, and no significant differences in the glucose disappearance rate (Kg) were observed at days 15, 45, 190, and 365 days after transplantation. The acute insulin response to glucose indicated no significant reduction of functional islet mass. CONCLUSIONS: PIT in severely insulinopenic type I diabetes mellitus primates resulted in restoration of normal glycemic parameters and durable islet mass. Operational tolerance was achieved with only 4 days of drug administration, sparing the animals from chronic exposure to potentially diabetogenic immunosuppressive drugs. These results offer an exciting new potential for type I diabetes mellitus treatment. PMID- 10670628 TI - IG-therasorb immunoapheresis in orthotopic xenotransplantation of baboons with landrace pig hearts. AB - BACKGROUND: The major problem of xenotransplantation is, that hyperacute xenograft rejection (HXR) causes graft failure within minutes or a few hours because of natural antibodies and activation of the complement system. As a preclinical model we transplanted pig hearts orthotopically into baboons. To prevent HXR after orthotopic xenotransplantation (oXHTx), the immunoglobulins (Ig) and natural antibodies were adsorbed to reusable Ig-Therasorb immunoadsorption (IA) columns. METHODS: We performed three oXHTx of landrace pig hearts into baboons (19+/-6.8 kg), using extracorporeal circulation (ECC) connected to the IA unit. After separating the recipient's blood into plasma and cellular fraction by a plasma filter, plasma flow was directed to the Ig Therasorb column coated with polyclonal sheep-antibodies against human IgG, IgM, and IgA. Intraoperative treatment consisted of 4 cycles of IA. For a control, we transplanted one pig heart into a baboon (16.9 kg) without applying IA. Perioperatively, serum concentrations of Ig, anti-pig-antibodies, complement and cardiac enzymes were determined. Tissue samples of myocardium were collected at the end of the study for immunohistochemical examinations, light microscopic examination (LM) and electron microscopic examination (EM). For cardiac monitoring after oXHTx, we used ECG, echocardiography, and invasive measurement of cardiac output. To prevent a mismatch of donor and recipient heart size, the donor pig had a 30-40% lower body weight than the recipient baboon. RESULTS: Four cycles of IA removed >80% of IgG, IgM, and IgA from plasma. The graft of the control animal failed after 29 min. The first oXHTx with IA was intentionally terminated after 100 min, the second oXHTx after 11 hr and the third oXHTx after 21 hr. All xenografts showed no histological signs of HXR. After weaning off ECC, these donor hearts worked in sinus rhythm without electrocardiographic ST-segment elevation. An excellent cardiac output was measured by echocardiography and thermodilution (2 L/min). Serological parameters indicating cardiac damage were significantly lower after IA if compared with the control experiment. Macroscopically, the xenograft of the control animal showed massive hemorrhage in comparison with the almost inconspicuous grafts after IA. The myocardium of the IA group demonstrated fewer deposits of Ig and complement components compared with the control animal. CONCLUSION: Baboons do not hyperacutely reject a porcine xenograft after antibody depletion by the Ig-Therasorb column. In our experiment only 4 cycles of immunoapheresis effectively prevented HXR after oXHTx of baboons. The Ig-Therasorb column is a reusable device, which can be handled easily in combination with the ECC. IA must be tested in oXHTx longterm survival experiments, especially in combination with transgenic pig organs, which could be a reliable preclinical approach for future clinical xenotransplantation. PMID- 10670629 TI - Hepatocyte growth factor is essential for amelioration of hyperglycemia in streptozotocin-induced diabetic mice receiving a marginal mass of intrahepatic islet grafts. AB - BACKGROUND: It is crucial for clinical islet transplantation to find a procedure to improve the success rate of insulin independence after islet transplantation. In the present study, we determined whether hepatocyte growth factor (HGF) has a favorable effect on amelioration of hyperglycemia in streptozotocin (STZ, 200 mg/kg)-induced diabetic mice (C57BL/6) receiving a marginal mass of intrahepatic islet isografts. METHODS: Isolated syngeneic islets were transplanted into the liver of recipients. HGF with dextran sulfate (DS) was administered intraperitoneally once a day at day 0, 2, 4, 6, and 8 relative to islet transplantation. DS has been known to enhance the effect of HGF. RESULTS: It was found that the number of 250 islets was a marginal mass as donor islets in this model, in which 2 out of 14 diabetic mice receiving 250 islets became normoglycemic by 90 days after transplantation. The treatment with HGF (100 microg) in conjunction with DS (200 microg) produced normoglycemia in all mice (n = 5). Morphological study as well as intraperitoneal glucose tolerance test revealed the beneficial effects of HGF. To our surprise, six out of nine mice receiving 250 islets and treated with DS alone became normoglycemic. Additional anti-HGF antibody treatment (100 microg, day -1, 0, 2, 4, 6, and 8) abolished the effects of DS, indicating that the effect by DS is mediated via the endogenous HGF. The effects of DS were not observed when the renal subcapsular space was the site of islet transplantation. There was a significant increase in plasma HGF levels in mice after the intrahepatic grafts but not the renal subcapsular one. CONCLUSIONS: These findings demonstrate that HGF is essential for amelioration of hyperglycemia in STZ-induced diabetic mice when a marginal mass of islets was grafted into the liver. As the liver is the site of clinical islet transplantation and the inability to achieve insulin independence after transplantation is a major obstacle for successful transplantation, HGF may facilitate to overcome such an important issue for clinical islet transplantation. PMID- 10670630 TI - Major histocompatibility complex class I peptide-pulsed host dendritic cells induce antigen-specific acquired thymic tolerance to islet cells. AB - BACKGROUND: As T-cell receptor-major histocompatibility complex (MHC) class I/self peptide interaction regulates T-cell development in the thymus, we reasoned that presentation of peptides by self dendritic cells (DC) to developing T cells in the thymus might induce acquired thymic tolerance. This hypothesis is based on the finding that intrathymic injection of allopeptides in the adult animal induces acquired tolerance. To examine this hypothesis, we studied the effects of intrathymic (IT) injection of a single immunodominant Wistar-Furth (WF) MHC class I (RT1.Au) peptide-pulsed host DC on islet allograft survival in the WF-to-ACI rat combination. METHODS: Bone marrow-derived ACI DC expressing MHC class I and II, OX62, and ED2 present allopeptides to naive and specifically peptide-primed syngeneic T cells in mixed lymphocyte reaction. Host DC pulsed with RT1.Au peptide 5 (residues 93-109) were injected into the thymus of streptozotocin-induced diabetic ACI that were transplanted 7 days later with donor-type (WF) or third-party (Brown Norway [BN]) islets. RESULTS: Whereas IT injection of 300 microg of peptide 5 alone led to normoglycemia and permanent islet survival in three of six diabetic ACI recipients, similar treatment combined with simultaneous intraperitoneal injection of 0.5 ml of anti-lymphocyte serum (ALS) on day -7 led to 100% permanent islet allograft survival (>200 days) compared to a mean survival time of 15.0+/-2.3 days in controls treated with ALS alone. In contrast, similarly prepared animals rejected the third-party (BN) islets in an acute fashion. To address the question of indirect allorecognition in acquired thymic tolerance, we examined the effect of peptide-pulsed host DC on graft survival. Whereas IT injection of peptide-pulsed host DC alone resulted in permanent islet survival in two of five animals, IT injection of peptide-pulsed host DC combined with 0.5 ml of ALS induced 100% donor-specific permanent islet allograft survival in the WF-to-ACI rat combination. These results suggest that thymic DC take up, process, and present the administered peptide to the developing T cells by the indirect allorecognition pathway in the induction of acquired thymic tolerance. CONCLUSION: We have demonstrated a novel approach to inducing transplant tolerance to islet allografts with IT injection of allopeptide-pulsed host DC. This finding suggests that immunization strategies using DC expressing MHC allopeptides or peptide analogue might be potentially useful in the treatment of autoimmune diabetes mellitus. PMID- 10670631 TI - NOX 100, a nitric oxide scavenger, enhances cardiac allograft survival and promotes long-term graft acceptance. AB - BACKGROUND: We examined the role of nitrosative stress in allograft destruction. METHODS: Rats undergoing cardiac transplants received NOX-100, a water-soluble nitric oxide (NO) scavenger with antioxidant properties, with or without low-dose cyclosporine (CsA). Graft survival, NO production, and nuclear factor kappa B (NF kappaB) activity were studied. RESULT: Using NOX-100 daily until rejection prolonged graft survival (11.6+/-0.6 vs. 7.4+/-0.2 days; P<0.05). Daily low-dose CsA (2.5 mg/kg im) for 7 days or until rejection also prolonged survival (12.6+/ 0.5 and 21.6+/-1.6 days, respectively; P<0.01 vs. Controls). Low-dose CsA for 7 days and NOX-100 for 30 days prolonged graft survival (45.0+/-4.7 days; P<0.01 vs. all groups.). NOX-100 had no effect on whole blood CsA levels. Combination therapy until Day 100 resulted in 1 graft loss at Day 116 and indefinite survival in 3 animals (>300 days), which accepted a second WF strain heart without further immunosuppressive therapy but promptly rejected a third party (ACI) cardiac allograft. NOX-100 and CsA reduced nitrate and nitrite, and combination therapy completely normalized NO through to Day 30. Electron paramagnetic resonance spectroscopic analysis demonstrated reduction of signals for nitrosylmyoglobin and nitrosyl-heme with NOX-100 and elimination of signals with CsA alone or combination therapy. Activity of myocardial NF-kappaB decreased with monotherapy vs. untreated allografts. Combination therapy resulted in further inhibition of NF-kappaB up to Day 30. The extent of graft survival correlated with the extent of NO scavenging and NF-kappaB inhibition. Short-term combination therapy had no effect on graft lymphocytic infiltrate on Days 15, 20, and 30. CONCLUSION: These data support a role for both oxidative and nitrosative stress in rejection and the immunoregulatory potential of antioxidant therapy after transplantation. PMID- 10670632 TI - Effect of surrogate tolerogenesis on the vascular rejection of pig heart xenografts. AB - BACKGROUND: Organ xenografts are fulminantly rejected by antibody-mediated vascular rejection. Surrogate tolerogenesis (ST), the induction of tolerance within the donor, is effective with aorta xenografts. This preliminary study assesses the effect of ST on preformed antibodies and rejection of porcine heart xenografts. METHODS: Tolerance to the donor pig was induced by infusing recipient marrow into fetal pigs. Later, pig splenocytes were transfused and heterotopic pig hearts transplanted using chimeric or nonchimeric pigs. Anti-pig antibodies were assessed. RESULTS: With ST alone, xenografts developed cellular rejection at 4-6 days, whereas control grafts developed vascular rejection at 3-4 days (cellular vs. vascular, P<0.03). There was a reduction in preformed antibodies (P<0.03). ST combined with moderate cyclosporine prevented rejection at 9+ and 25 days in sensitized recipients compared with vascular rejection at 0.5-2 days for controls (P<0.07). CONCLUSIONS: ST seems to provide protection against vascular rejection. The cellular rejection seems sensitive to cyclosporine. PMID- 10670633 TI - Prolongation of canine liver allograft survival by a novel immunosuppressant, FTY720: effect of monotherapy and combined treatment with conventional drugs. AB - BACKGROUND: The immunosuppressive effect and other properties of a novel immunosuppressant, FTY720, have been studied mostly in the experimental transplantation of various extrahepatic organs. In this experiment, we evaluated the antirejection potency and adverse effects of this agent on liver grafts using a canine liver transplantation model. METHODS: Forty-eight orthotopic liver transplantations were performed by the standard technique under a veno-venous bypass. Liver recipients were divided into two studies: a single-dose study with FTY720 at various doses and a combined dose study with conventional immunosuppressants (cyclosporine or tacrolimus) alone and combined with FTY720. Survival, biochemical and hematological tests, blood levels of immunosuppressants, and postmortem histology were determined. RESULTS: The median survival of untreated control animals was 9 days, whereas treatment with FTY720 at a dose of 0.1 mg/kg/day prolonged graft survival to 49.5 days. FTY720 at 1 mg/kg/day showed a slight but insignificant prolongation to 16 days, but when the dose was increased to 5 mg/kg/day, the graft was rejected at 10 days. The combination of FTY720, 0.1 mg/kg/day, with a subtherapeutic dose of cyclosporine, 5 mg/kg/ day, prolonged median animal survival from 40 days with cyclosporine alone to 74 days. A combination of FTY720 (0.1 mg/kg/day) with tacrolimus (0.5 mg/kg/ day) compromised animal survival, reducing survival from 83.5 days with tacrolimus alone to 30.5 days due to infectious complication and emaciation by overimmunosuppression. No evident drug-induced side effects were observed. CONCLUSIONS: FTY720 has a potent immunosuppressive effect when used alone at 0.1 mg/kg/day in canine liver transplantation. FTY720 is a promising candidate for future clinical application in orthotopic liver transplantation. PMID- 10670634 TI - Bile acids in xenogeneic ex-vivo liver perfusion: function of xenoperfused livers and compatibility with human bile salts and porcine livers. AB - BACKGROUND: In recent years, hepatic support systems using xenogeneic cells have been developed to support patients in fulminant hepatic failure. The extent to which xenogeneic hepatocytes metabolize and excrete human organic anions is unclear. In these studies we examined the ability of the ex vivo porcine liver to clear human bile acids during extracorporeal liver perfusion (ELP). METHODS: Four patients with fulminant hepatic failure underwent extracorporeal liver perfusion with 9 porcine livers. The venovenous circuit was designed as previously described (NEJM,1994,331:234) as were the immunologic features (Transplantation 1994,58:1162). Bile from the porcine liver and serum samples were collected hourly during perfusion. Three bile acids (glycocholic, glycodeoxycholic, taurodeoxycholic acid) were selected as markers for human bile and three (glycohyocholic, glycohyodeoxycholic, and glyco-3alpha-hydroxy-6-oxo-5beta cholanoic acid) for markers of pig bile. Bile acids from both serum and bile were processed and analyzed through high performance liquid chromatography. The Students' t test was used for statistical analysis. RESULTS: The mean duration of perfusions was 4.1+/-1.5 hr. The mean total bile acid clearance from serum (243+/ 44 micromol/h) was similar to the total bile acid biliary excretion (286+/-84 micromol/hr, P = 0.06). After 1 hr of perfusion, bile samples demonstrated a predominance of pig bile salts (65%). After 3 hr of perfusion, human bile acids made up 85% of total biliary bile acids. Pig bile acids appeared in patients' sera after 1 hr of perfusion, and after 3 hr, 35% of serum bile salts were pig specific. CONCLUSIONS: Porcine livers perfused with human blood can clear the serum of potentially toxic human bile acids and excrete them into bile. Simultaneously, the percentage of pig-specific bile acids in patient serum increases during xenogeneic perfusion for unknown reasons. The relative hepatic uptake of bile acid from serum is similar to bile acid excretion in bile. Further development of systems using porcine livers or hepatocytes is warranted. PMID- 10670635 TI - The influence of pulsatile preservation on renal transplantation in the 1990s. AB - BACKGROUND: Unlike simple cold storage (CS), pulsatile machine preservation (MP) of kidneys for transplantation permits pharmacologic manipulation of the perfusate and aids in the pretransplant assessment of the kidney graft. These characteristics of MP may have importance in the era of increasing use of extended criteria donor kidneys. The overall aim of this article is to critically assess practices at our preservation unit with respect to graft function. Specific aims are to (1) compare the influence of MP versus CS on graft function, (2) determine which pretransplant variables have significance in pretransplant assessment, and (3) determine whether pharmacologic manipulation during MP is advantageous. METHODS: There were 650 consecutive kidneys preserved in our laboratory between January 1, 1993 and March 1, 999, by either MP or CS. All MP kidneys were preserved by continuous hypothermic pulsatile perfusion using Belzer MPS or Belzer II solution. Perfusion parameters and electrolytes were measured serially during pulsatile perfusion. All CS kidneys were stored in University of Wisconsin solution. All kidneys obtained from donors exhibiting extended criteria features underwent pretransplant frozen section biopsies. Transmission electron microscopy (EM) was performed on a subset of kidneys undergoing pharmacologic manipulation. Four agents were assessed prospectively for their ability to influence MP characteristics when added to perfusate: PGE1, trifluoperazine, verapamil, and papaverine. RESULTS: MP was associated with improved immediate, 1 , and 2-year graft function and reduced length of initial hospital stay when compared with CS grafts. Changes in the machine perfusion variables flow and resistance, and the [Ca++] in perfusate, were significantly associated with delayed graft function (DGF) after the transplant. Biopsy information was not predictive of DGF. The addition of PGE1 to perfusate improved MP characteristics, reduced the release of [Ca++] into perfusate, and ameliorated mitochondrial ischemic injury in transmission EM images. Early graft function was improved in the presence of PGE1+MP, compared with function in the presence of other pharmacologic agents or CS alone. CONCLUSIONS: MP is associated with improved early and long term renal function. Moreover, PGE1 augments MP in improving graft function. The combination of MP+PGE1 may be important in optimizing the ability to use extended donor criteria kidneys and, thereby, improve the overall efficiency of cadaveric renal transplantation. PMID- 10670636 TI - Right lobe graft in living donor liver transplantation. AB - BACKGROUND: For the sake of donor safety in living donor liver transplantation (LDLT), the left lobe is currently being used most often for the graft. However, size mismatch has been a major obstacle for an expansion of the indication for LDLT to larger-size recipients, because a left lobe graft is not safe enough for them. METHODS: In 1998, LDLT using a right lobe graft was introduced and performed on 26 recipients to overcome the small-for-size problem. The right lobe, which does not include the middle hepatic vein of the donor, was used. Initially, indication for right lobe LDLT was basically defined as an estimated left lobe graft volume/recipient body weight ratio (GRWR) of <0.8%, which was later raised to <1.0%. RESULTS: All the donors recovered from the operation without persistent complications. Two donors with transient bile leakage were successfully treated with a conservative approach. A right lobectomy resulted in more blood loss (337+/-175 ml), and a longer operative time (6.67+/-0.85 hr) than a lateral segmentectomy, but not a left lobectomy. Grafts with a GRWR >0.8% were implanted in all recipients, except for two, who received relatively smaller right lobes (GRWR of 0.68% and 0.66%). In one of these two, the right lobe from the donor was used as the orthotopic auxiliary graft. Postoperative transitory increases in total bilirubin and aspartate transaminoferase for right lobe donors were higher than those for the left lateral segmentectomy. Nineteen recipients (73.1%) were successfully treated with this procedure. The causes of death were not specific for right lobe LDLT, except for one patient with a graft that had multiple hepatic venous orifices. These multiple and separate anastomoses of the hepatic veins caused an outflow block as a result of a positional shift of the graft, which finally led to graft loss. CONCLUSION: Our experience suggests that right lobe grafting is a safe and effective procedure, resulting in the expansion of the indication for LDLT to large-size recipients. How to deal with the possible variation in the anatomy of the right lobe graft should be given attention throughout the procedure. PMID- 10670637 TI - Results of pancreas transplantation after steroid withdrawal under tacrolimus immunosuppression. AB - PURPOSE: The results of steroid withdrawal in pancreas transplant recipients under tacrolimus immunosuppression were analyzed. METHODS: From July 4, 1994 until April 30, 1998, 147 pancreas transplantations were performed in 141 patients, including 126 simultaneous pancreas-kidney transplantations, 13 pancreas after kidney transplantation, and 8 pancreas transplantations alone. Baseline immunosuppression consisted of tacrolimus and steroids without antilymphocyte induction. Twenty-three patients were excluded from analysis because of early graft loss in 17 cases, retransplantation in 5 cases, and simultaneous pancreas-kidney transplantation after heart transplantation in 1 patient. RESULTS: With a mean follow-up of 2.8+/-1.1 years (range 1.0 to 4.8 years), complete steroid withdrawal was achieved in 58 (47%) patients with a mean time to steroid withdrawal of 15.2+/-8 months (range 4 to 40 months after transplantation). Of the entire cohort of 141 patients, overall 1-, 2-, and 4 year patient survival rates were 98%, 95.5%, and 86%, respectively. Overall 1-, 2 , and 4-year graft survival rates were 83%, 80%, and 71% (pancreas) and 95%, 91%, and 84% (kidney), respectively. Of the 124 patients analyzed for steroid withdrawal, 1-, 2-, and 4-year patient survival rates were 98%, 97%, and 92%, respectively. Overall 1-, 2-, and 4-year graft survival rates were 98%, 91.5%, 83% (pancreas) and 97%, 95%, and 91% (kidney). Patient, pancreas, and kidney survival rates at 1 year were 100%, 100%, and 98% (off steroids) versus 97%, 91%, and 96% (on steroids, all NS) and at 4 years were 100%, 94%, and 95% (off steroids) versus 78%, 68%, and 85% (on steroids, P = 0.01, 0.002, and NS, respectively). The cumulative risk of rejection at the time of follow-up was 76% for patients on steroids versus 74% for patients off steroids (P = NS). Seven patients originally tapered off steroids were treated for subsequent rejection episodes, which were all steroid sensitive, and two of these seven patients are currently off steroids. Thirteen patients received antilymphocyte therapy for steroid-resistant rejection, five of whom are now off steroids. Tacrolimus trough levels were 9.3+/-2.4 ng/ml (off steroids) and 9.7+/-4.3 (on steroids, P = NS). Mean fasting glucose levels were 98+/-34 mg/dl (off steroids) and 110+/-41 mg/dl (on steroids, P = NS). Mean glycosylated hemoglobin levels were 5.2+/-0.9% (off steroids) and 6.2+/-2.1% (on steroids, P = 0.02), and mean serum creatinine levels were 1.4+/-0.8 mg/dl (off steroids) and 1.7+/-1.0 mg/dl (on steroids, P = 0.02). CONCLUSION: These data show for the first time that steroid withdrawal can be safely accomplished in pancreas transplant recipients maintained on tacrolimus based immunosuppression. Steroid withdrawal is associated with excellent patient and graft survival with no increase in the cumulative risk of rejection. PMID- 10670638 TI - Liver allotransplantation after extracorporeal hepatic support with transgenic (hCD55/hCD59) porcine livers: clinical results and lack of pig-to-human transmission of the porcine endogenous retrovirus. AB - BACKGROUND: Whole organ extracorporeal perfusion of a genetically modified humanized (transgenic) pig liver has been proposed as a technology that may sustain patients with severe liver failure while awaiting human liver transplantation. METHODS: We report on two cases of successful extracorporeal perfusion of a transgenic pig liver in patients awaiting transplantation for fulminant hepatic failure. The pig livers used were transgenic for human CD55 (decay-accelerating factor) and human CD59. These transgenic modifications are designed to reduce or eliminate the hyperacute rejection inherent in pig-to primate xenotransplants. We also report on the results of serial surveillance testing for presence of the porcine endogenous retrovirus (PoERV) in these two patients. RESULTS: Extracorporeal perfusion in two patients was performed for 6.5 and 10 hr, respectively, followed by the successful transplantation of a human liver and resultant healthy patients (18 and 5 months later as of this writing). The porcine livers showed evidence of synthetic and secretory function (decreasing protime and bilirubin, bile production). Serial polymerase chain reaction analysis of these patients' peripheral blood mononuclear cells has failed to show presence of PoERV DNA sequences. CONCLUSIONS: The CD55/CD59 transgenic porcine liver appears capable of safely "bridging" a patient to liver transplantation. Human PoERV infection from these livers has yet to be demonstrated. PMID- 10670639 TI - Outcome in recipients of dual kidney transplants: an analysis of the dual registry patients. AB - BACKGROUND: A novel but controversial method to increase the utilization of aged donor kidneys is the transplantation of both kidneys as a dual transplant. Initial single-center reports demonstrated outcomes similar to single kidneys from younger donors. In this report, we compare outcome in recipients of kidneys from donors > or =54 years of age who received a single kidney transplant reported to the United Network for Organ Sharing Scientific Registry versus a dual kidney transplant reported to the Dual Kidney Registry. METHODS: A retrospective analysis was performed, comparing four donor and nine recipient and outcome variables between recipients of a single versus a dual transplant between March 1993 and March 1999. RESULTS: Dual versus single transplants from donors > or =54 years of age have a significantly decreased incidence of delayed graft function, and lower serum creatinines up to 2 years after transplant despite having kidneys from significantly older donors with poorer HLA matching. CONCLUSIONS: Dual kidney transplants improve graft performance and outcome in recipients of kidneys from donors > or =54 years of age. PMID- 10670640 TI - The kinetics of tolerance induction by nondepleting anti-CD4 monoclonal antibody (RIB 5/2) plus intravenous donor alloantigen administration. AB - BACKGROUND: CD4+ T cells play an essential role in allograft rejection. The monoclonal anti-rat CD4 antibody, RIB 5/2, has been shown to modulate the CD4 glycoprotein without eliminating the recipient T cells. We have successfully induced tolerance to rat heart allografts by recipient pretreatment with a single dose of RIB 5/2 plus intravenous administration of donor splenocytes. In this study, we explored whether this potent regimen could induce tolerance to the more resistant kidney and skin allografts. Furthermore, we examined the kinetics and requirements for tolerance to be met by a single dose of RIB 5/2 plus i.v. alloantigen. METHODS: The efficacy of a single i.p. dose of 20 mg/kg RIB 5/2 plus i.v. donor antigen (25x10(6) splenocyte) pretreatment 0, 21, or 40 days before receipt of an MHC-mismatched Lewis (RT1l) to Buffalo (RT1b) rat cardiac, renal, or skin allograft was studied. Another group of Buffalo recipients treated with RIB 5/2 plus an i.v. alloantigen +/-thymectomy received kidney transplants after 40 days. Attempts to prevent tolerance used interleukin-2 or prior sensitization. Mixed lymphocyte cultures, cytotoxic assays, and precursor frequencies of helper and cytotoxic cells, by limiting dilution analysis, serially measured in vitro cell-mediated immunity. RESULTS: RIB 5/2 administration combined with i.v. alloantigen 21 days before induced tolerance to heart and kidney allografts but did not prolong skin graft survival. In contrast, kidney allografts delayed for 40 days after pretreatment were acutely rejected and survival was not affected by the thymectomy. MLC, CTL, and pTH, and pCTL precursor frequencies from recipients of long-term grafts were specifically suppressed to donor, but not third party, alloantigen. CONCLUSION: A single dose of the nondepleting anti-CD4 monoclonal antibody, RIB 5/2, plus i.v. alloantigen is a potent inducer of tolerance to heart and kidney, but not skin, allografts. The RIB 5/2-induced donor unresponsiveness to a delayed kidney or cardiac allograft is time dependent but can be prolonged if specific alloantigen is present. Suppression of cell-mediated allo-immune responsiveness correlates with allograft acceptance. PMID- 10670641 TI - Brief cyclosporine treatment prevents intrathymic (IT) tolerance induction and precipitates acute rejection in an IT rat cardiac allograft model. AB - BACKGROUND: Intrathymic (IT) alloantigen combined with administration of rabbit anti-rat anti-lymphocyte serum (ALS) intraperitoneally induces donor-specific tolerance to rat cardiac transplants. The purpose of this study was to examine the effect of a brief course (4 days) of cyclosporine (CsA) on the development of IT tolerance. METHODS: Buffalo (BUF) (RT1b) rats were given 25x10(6) fully MHC mismatched Lewis (LEW) (RT1l) splenocytes by IT injection plus 1.0 ml of ALS intraperitoneally. Twenty-one days later, IT donor-specific LEW (group 1) or third-party (ACI, RT1a) (group 2) hearts were heterotopically transplanted to the abdominal aorta A third group of BUF (group 3) were given daily CsA (10 mg/kg) by oral gavage for 4 days before administration of IT LEW cells and ALS. Rejection as defined by the cessation of a palpable heartbeat was confirmed by histology. Cytokine profiles of allografts from all groups were then analyzed using a multi probe RNase protection assay. RESULTS: Sixty-seven percent of IT/ALS-treated BUF recipients not pretreated with CsA accepted LEW heart grafts for greater than 90 days. However, 86% of animals treated with CsA for 4 days before IT injection and ALS rejected allografts at 10.7+/-3.2 days. Third-party allografts (ACI) were uniformly rejected (7.0+/-0.0 days). Histology confirmed cellular rejection in CsA-treated allografts and cytokine analysis detected increased interleukin (IL) 3, IL-5, and tumor necrosis factor-alpha when compared to increased IL-2 and interferon-gamma in rejecting untreated controls. CONCLUSIONS: CsA can prevent the induction of intrathymic alloantigen tolerance. These results support the development of a CsA-sensitive, but IL-2-independent, active regulatory mechanism after intrathymic exposure to donor-specific alloantigen and depletion of mature peripheral T cells. PMID- 10670642 TI - Both ischemic and pharmacological preconditioning decrease hepatic leukocyte/endothelial cell interactions. AB - BACKGROUND: Ischemic preconditioning has been shown to protect some tissues from ischemia/reperfusion (I/R) injury. Adenosine is believed to play an important role by attenuating leukocyte-endothelial cell adhesive interactions. Dipyridamole increases adenosine bioavailability. The purpose of this study was to evaluate the effects of mechanical (MPC) and pharmacological preconditioning (PPC) on leukocyte endothelial cell interaction in hepatic I/R injury. METHODS: C57BL6 mice were subjected to 30 min of ischemia to the left lobe of the liver. Groups tested at 30 min, 2, 5, 12, and 24 hr of reperfusion had 1) sham laparotomy (n = 10, 2) I/R (n = 25), 3) ischemic preconditioning with 5 min of ischemia and 10 min reperfusion before I/R (n = 25), and 4) (PPC) with dipyridamole (n = 25). Intravital microscopic examination was used to assess leukocyte/endothelial cell adhesion. Blood was drawn for leukocyte counts and liver function tests. RESULTS: A significant decrease in leukocyte rolling was observed at 30-min and 5-hr reperfusion intervals in the PPC and ischemic preconditioning groups compared with the I/R group. A significant decrease in leukocyte saltation was also observed in the PPC and MPC groups at 2, 5, and 12 hr of reperfusion when compared with the I/R group. aspartate aminotransferase was significantly decreased in the 5-hr preconditioning groups. There was not a significant decrease in the white blood cell count because of PPC or MPC vs. I/R CONCLUSIONS: Preconditioning decreases endothelial/ leukocyte interaction and reduces liver damage as measured by aspartate aminotransferase. These data prove that IPC and PPC provide some degree of hepatic protection in I/R injury. PMID- 10670643 TI - Defining hepatocellular chimerism in a liver failure patient bridged with hepatocyte infusion. AB - BACKGROUND: A practical method of monitoring engraftment by transplanted hepatocytes for the purpose of bridging human liver failure to native regeneration is described. METHODS: A previously healthy 37-year-old female with a 2-week history of a febrile illness presented with fulminant liver failure. Findings on admission included the following: illicit drug use, serum hepatitis B surface antigen positive, grade 1 encephalopathy, prothrombin time (pt) >100 sec, F7<1%, NH3 150 micromol/L, alanine aminotransferase 4079 U/L, total bilirubin level 11.4 mg/dl, and glucose 70 mg/dl (on IV D10). With immunosuppression, 8.8x10(8), 96% viable human hepatocytes were intraportally infused. Clinical chemistries, total sHLA class I, and ELISA to measure donor-specific sHLA-A1 and B8 were recorded. Serial transjugular liver biopsies were performed and pooled for histological examination, DNA extraction, and HLA DNA typing. RESULTS: The patient fully recovered. At months 3 and 4 with donor biopsy specimen class I HLA DNA no longer detectable, immunosuppression was tapered off. The patient is clinically normal, serum hepatitis B surface antigen negative at 10 months of follow-up. CONCLUSIONS: Bridging liver failure with donor hepatocytes with HLA class I antigen disparate from recipients is clinically feasible, and allows for a marker, combined with serial graft histology, to safely wean immunosuppression when native liver regeneration succeeds. PMID- 10670644 TI - Experience with daclizumab in liver transplantation: renal transplant dosing without calcineurin inhibitors is insufficient to prevent acute rejection in liver transplantation. AB - BACKGROUND: Daclizumab is a monoclonal antibody directed against the alpha chain of the interleukin 2 receptor. We review our experience with the use of daclizumab in liver transplant recipients. METHODS: Thirty-two patients were given daclizumab as induction therapy in the setting of hepatic transplantation. Seven of these patients were enrolled in a pilot study to determine the efficacy of daclizumab in conjunction with corticosteroids and mycophenolate mofetil without the initial use of calcineurin inhibitors (CI). The remaining 25 patients received daclizumab, mycophenolate mofetil, and steroids, with the institution of CI generally within the first postoperative week. The majority of these patients (n = 17) had some degree of renal insufficiency. RESULTS: The pilot study was halted after the first seven patients were enrolled because of an unacceptably high rate of rejection (7/7 = 100%). The patients outside of this pilot study, however, had a much lower rate of rejection (36%). The incidence and severity of rejection correlated with the delay in institution of CI. The described dosing schedule resulted in subtherapeutic daclizumab levels in liver transplant recipients. CONCLUSIONS: Daclizumab used in liver transplant recipients without any CI was ineffective and can potentially lead to steroid-resistant rejection. The dosing regimen used in renal transplant recipients is most likely insufficient for liver transplant patients. However, daclizumab can be used safely in patients with preexisting or postoperative renal dysfunction in conjunction with low doses of CI given within the first week postoperatively. PMID- 10670645 TI - Cadaveric versus living donor kidney transplantation: a Medicare payment analysis. AB - BACKGROUND: We found previously that the clinical advantages of living donor (LD) renal transplantation lead to financial cost savings compared to either cadaveric donation (CAD) or dialysis. Here, we analyze the sources of the cost savings of LD versus CAD kidney transplantation. METHODS: We used United States Renal Data System data to merge United Network for Organ Sharing registry information with Medicare claims data for 1991-1996. Information was available for 42,868 CAD and 13,754 LD transplants. More than 5 million Medicare payment records were analyzed. We calculated the difference in average payments made by Medicare for CAD and LD for services provided during the first posttransplant year. RESULTS: Average total payments were $39,534 and $24,652 for CAD and LD, respectively (P<0.0001) during the first posttransplant year. The largest source of the difference in payments was in inpatient hospitals, representing $10,653.67 (P<0.0001). For patients who had Medicare as the primary payer, average transplant charges were significantly higher for CAD donation ($79,730 vs. $69,547, P<0.0001); average transplant payments demonstrated no statistical differences ($28,483 vs. $28,447, P = 0.858). Therefore, inferred profitability was significantly higher for LD. CONCLUSIONS: Medicare payments are remarkably lower for LD compared to CAD in every category. The single largest cost saving comes from inpatient hospital services. A portion of the savings from LD could be invested in programs to expand living kidney donation. PMID- 10670646 TI - Influence of surgical procedures on interleukin-6 and monocyte chemotactic and activating factor responses: CABG vs. valvular surgery. AB - Interleukin-6 (IL-6) and monocyte chemotactic and activating factor/monocyte chemoattractant protein-1 (MCAF/MCP-1) play pivotal roles in systemic inflammation, immune response, and tissue damage after cardiopulmonary bypass (CPB). Previous reports have described transient rises in IL-6 and MCAF after CPB, but the data seem to vary according to the different surgical procedures used. To evaluate the influence of the different surgical procedures on the proinflammatory cytokine responses, we compared perioperative serum IL-6 and MCAF release in coronary artery bypass grafting (CABG) and valvular surgery cases. Eighteen CABG (CABG group) and 7 single valvular cardiac surgery patients (valve group) were included in this study. Blood samples were taken to measure the serum concentrations of IL-6 at the induction of anesthesia, at the removal of the aortic cross-clamp, at the end of CPB, at the end of surgery, and 24 h after the termination of surgery. Serum IL-6 and MCAF were assayed by ELISA. Serum IL-6 increased immediately after aortic declamping and reached its peak at the end of surgery in both groups. Serum IL-6 concentrations at the end of surgery and 24 h after surgery were significantly higher in the valve group than in the CABG group (123.9 +/- 21.7 pg/ml vs. 79.7 +/- 10.4 pg/ml, p = 0.049; 113.6 +/- 25.0 pg/ml vs. 39.9 +/- 11.5 pg/ml, p = 0.006, respectively). Serum MCAF increased immediately after aortic declamping, and the MCAF level at the end of surgery was significantly higher in the valve group than in the CABG group (1118.4 +/- 353.9 pg/ml vs. 241.0 +/- 71.2 pg/ml, p = 0.002, respectively). IL-6 and MCAF may play important roles in the pathophysiology of surgical damage with CPB, and the different surgical procedures appear to affect the proinflammatory cytokine release after cardiac surgery differently. PMID- 10670647 TI - Orthodontic movement induces high numbers of cells expressing IFN-gamma at mRNA and protein levels. AB - Cytokines are important signaling proteins that are liberated during immune challenges and exhibit many modulatory activities. However, their role in periodontal modeling during orthodontic tooth movement is not fully understood. The aim of this study was to analyze effects of mechanical force during orthodontic tooth movement, in the pressure zone, on the induction of interferon gamma (IFN-gamma) as a proinflammatory cytokine of Th1 type and interleukin-4 (IL 4)/IL-10 as anti-inflammatory cytokines of Th2 type. In 12 Wistar rats 40-45 days old, the maxillary first molar was moved mesially by means of a closed coil spring for 3, 7, and 10 days. The contralateral side served as a control. IFN gamma, IL-4, and IL-10 mRNA were determined by in situ, hybridization, and protein levels of IFN-gamma was measured by immunohistochemistry. Induction of IFN-gamma at both mRNA and protein levels was significantly higher on the experimental side than on the contralateral control side on day 3. The signal gradually became stronger on day 7 and remained high on day 10. Cytokines of the Th2 type (IL-4 and IL-10) were not detected at all examined time points in both pressure and contralateral control sides. Considering the potential immunoregulatory roles played by IFN-gamma, our data suggest that IFN-gamma may be involved in periodontium remodeling during orthodontic tooth movement. PMID- 10670648 TI - A Stat1alpha factor regulates the expression of the human vimentin gene by IFN gamma. AB - Vimentin is an intermediate filament protein normally expressed in cells of mesenchymal origin. Here, we report an increase in vimentin gene transcription induced by the cytokine interferon-y (IFN-gamma). Northern blot analysis and reporter gene assays reveal that IFN-gamma induces vimentin gene transcription in HeLa cells. However, no increase in vimentin mRNA synthesis was observed de novo in MCF-7 cells, which do not already express vimentin. Band shift analysis shows that the Stat1alpha protein mediates vimentin induction by IFN-gamma. A human mutant fibroblast cell line (U3A), which lacks Stat1alpha but expresses vimentin mRNA, yields no increase in vimentin mRNA levels on the addition of IFN-gamma. These results suggest that the induction of vimentin gene expression might be an important part of a complex cellular response to IFN-gamma. PMID- 10670649 TI - Transduction and utility of the granulocyte-macrophage colony-stimulating factor gene into monocytes and dendritic cells by adeno-associated virus. AB - The genetic manipulation of antigen-presenting dendritic cells (DC) offers promise for stimulating the immune response, in particular for anticancer and antiviral protocols. As adeno-associated virus (AAV) has shown promise as a gene delivery vector for transducing a variety of hematopoietic cell types, we have investigated AAV's ability to genetically alter DC. In this analysis, we modified the standard granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) treatment of adherent monocytes to generate DC. In our protocol, adherent monocytes were first infected with an AAV/GM-CSF/Neo vector, and the addition of IL-4 was delayed for 2 days to allow for a brief period of monocyte proliferation. AAV-mediated transduction of the GM-CSF and Neo genes into monocytes/DC precursors was demonstrated by G418 selection, GM-CSF secretion, GM-CSF RNA expression (reverse transcriptase-polymerase chain reaction amplification [RT-PCR]), and cell proliferation. Cells resulting from infection with AAV/GM-CSF/Neo virus, and subsequent IL-4 and tumor necrosis factor-alpha (TNF-alpha) treatment, displayed multiple classic markers consistent with mature DC. Finally, chromosomal integration of the AAV vector was also demonstrated in sorted CD83+ DC. These data strongly suggest that AAV vectors will be useful for the genetic manipulation of DC and suggest that the transduction of the GM-CSF gene was able to fully replace the need for exogenous GM-CSF in the production of mature DC. PMID- 10670650 TI - Restoration of cytotoxic T lymphocyte function in malignant pleural effusion: interleukin-15 vs. interleukin-2. AB - The present study attempts to define the role of interleukin-15 (IL-15), as compared with IL-2, in generating cytotoxic T lymphocytes (CTL) from the malignant effusions of cancer patients. Effusion-associated lymphocytes (EAL) from malignant effusion were incubated with IL-15 or IL-2 with or without alphaCD3. Proliferation and cytotoxicity assays were performed. IL-15 was found to have at least an equivalent, if not higher, activity to IL-2 in terms of lymphocyte proliferation and generation of CTL from EAL. The proliferative response of EAL, cocultured with IL-15, with or without alphaCD3, was partly inhibited by pretreatment with an anti-IL2 receptor beta chain monoclonal antibody (mAb). The proliferative response of EAL, cocultured with alphaCD3, IL 2, or both, was partly inhibited by pretreatment with an anti-IL-2 receptor alpha chain mAb. Overnight [5lCr] release assays against K562, Daudi, and the patients' autologous tumor cells were done to evaluate EAL's cytolytic activity. MHC class I Ab blocked the stimulated cytolytic activity of EAL against autologous tumors. An mAb depletion assay showed that the phenotype of the restored EAL was CD16-CD4 CD8+; thus, the restored activity of EAL was CTL activity. The results suggest that both IL-15 and IL-2 can restore CTL activity from EAL in the presence of T cell receptor (TCR)-CD3 engagement, but the effect of IL-15 was superior. PMID- 10670651 TI - Acute-phase proteins and hematologic values in ovine lentivirus-infected lambs treated with recombinant ovine IFN-tau. AB - To evaluate changes in complete blood cell (CBC) counts, haptoglobin and fibrinogen in ovine lentivirus (OvLV)-infected lambs treated with recombinant ovine interferon-tau (rOVIFN-tau), 24 lambs were allocated to one of four groups (n = 6 per group): (1) virus + rOvIFN-tau, VI, (2) virus + placebo, VP, (3) no virus + rOVIFN-tau, NVI, and (4) no virus + placebo, NVP. Three lambs in each group were treated once a day for 12 weeks, and the remaining 3 lambs were treated for 33 weeks. Blood was collected at days 0, 7, and 10 and at weeks 2-10, 12, 32, and 33 to determine CBC counts, as well as haptoglobin and fibrinogen levels. Hematologic values remained within normal limits in all groups. However, hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and packed cell volume (PCV) values decreased (p < 0.05) in the two rOvIFN-tau-treated groups (VI and NVI) compared with the placebo-treated (VP and NVP) groups. Both rOvIFN-upsilon and OvLV had a mild negative effect on neutrophil numbers. Although Hb, MCV, MCHC, PCV, and neutrophil values declined in the rOvIFN-tau-treated lambs compared with the placebo-treated lambs, these values remained within the reference range for sheep. Experimental lambs did not show adverse clinical signs associated with OvLV infection or as a result of rOvIFN-tau treatment. The lack of significant side effects of high-dose rOvIFN tau in sheep and previous reports of broad-spectrum and cross-species antiviral activity suggest that rOvIFN-tau warrants further investigation as an antiviral therapeutic agent. PMID- 10670652 TI - Characterization of the cytokine network at a single cell level in mice with collagen-induced arthritis using a dual color ELISPOT assay. AB - Collagen-induced arthritis (CIA) in mice has been classified as a Thl-mediated disease. However, most evidence for this has been obtained by indirect experiments; for example, the administration of neutralizing anti-interferon gamma (IFN-gamma) antibody reduced the severity of arthritis. To obtain direct evidence about the cytokine balance in CIA mice, we analyzed the cytokine secreting cell in CIA mice at the single cell level using a dual color enzyme linked immunospot (ELISPOT) assay, which enabled us to analyze interleukin-2 (IL 2)-secreting cells or IL-4-secreting cells or both simultaneously. Furthermore, to characterize the cytokine network in the pathogenesis of CIA, the frequency of the cells secreting IL-12, which induced the development of naive Th cells into Th1 cells, was analyzed. The results show that in the prearthritic phase, the number of IL-12-secreting cells in spleen and peritoneal exuded cells is increased, and Th1 cells in lymph node and spleen are dominant. In contrast, after the onset of clinical arthritis, the number of IL-12-secreting cells in spleen, lymph node, and peritoneal exuded cells is decreased, and there is a shift from a Thl-dominant to a Th2-dominant state in lymph node and spleen. The results indicated that the pathogenesis of CIA is associated with a disruption in the normal ratio of Th1/Th2 at cell level. PMID- 10670653 TI - The capacity to produce IFN-gamma rather than the presence of interleukin-4 determines the resistance and the degree of susceptibility to Leishmania donovani infection in mice. AB - The immune response against Leishmania donovani infection has been investigated in one resistant mouse strain (C3H/HeJ) and three susceptible mouse strains (C57BL/6, BALB/c, and B10D2/n). In order to correlate the strain-specific course of infection with the individual T cell response phenotype, the ex vivo cytokine secretion patterns of splenic lymphocytes were assessed by ELISA (interferon-y [IFN-gamma], interleukin-4 [IL-4], IL-10) or by bioassay (IL-2). The strain dependent differences in the course of infection correlated closely with the potency of T cells to produce IFN-gamma. C3H/HeJ mice produced high amounts of IFN-gamma before and during infection, whereas susceptible mice produced low amounts of IFN-gamma early during L. donovani infection. However, C57BL/6 mice, which recovered from the infection rapidly after the acute stage, developed marked IFN-gamma response within the first 30 days of infection. In contrast, in BALB/c and B10D2/n mice, the IFN-gamma production diminished during the acute stage, and this was associated with a delay in recovery and with subsequent switching into the chronic stage. Interestingly, CD8+ T cells contributed significantly to IFN-gamma production during this phase. In contrast to IFN-y, the levels of IL-4 in response to antigen or mitogen ex vivo were always very low. Moreover, neutralization of endogenous IL-4 in vivo by treatment with soluble murine IL-4 receptor did not result in significant decreases in the parasite burdens in spleen and liver but did cause a decrease in the serum IgE level of L. donovani-infected BALB/c mice. These results confirm that in visceral leishmaniasis a Thl-dominated immune response is protective against the L. donovani parasites and, furthermore, that the capacity to produce IFN-gamma rather than the presence of IL-4 determines the efficacy of the immune response in susceptible mice. The data show that CD8+ T cells represent an important source of IFN-gamma during L. donovani infection in susceptible mice, implying a role for this cell type in healing and development of protective immunity. PMID- 10670654 TI - A sensitive and versatile bioassay for ligands that signal through receptor clustering. AB - The induced expression of xanthine-guanine phosphoribosyl transferase (XGPRT) by low concentrations (-2 pg/ml) of interferon-alpha (IFN-alpha) or IFN-beta, in the 2fTPGH cell line caused a 50% cytotoxicity when these cells were grown in medium containing 6-thioguanine. We extended the application of this sensitive, reliable, and easy bioassay to other members of the cytokine family. To activate the IFN signaling pathway, we made receptor chimeras, consisting of the IFN type I receptor intracellular and transmembrane domains, fused to either the interleukin-5 (IL-5) receptors or erythropoietin (Epo) receptor extracellular domains as model systems. 2fTGH cells, stably transfected with these receptor chimeras, responded to very low concentrations of IL-5 or Epo (IC50 values of approximately 15 pg and 3 pg/ml, respectively) and thus can be used as a very sensitive bioassay for both ligands. Background activity of IL-5, Epo, tumor necrosis factor (TNF), IL-6, or leptin on cells that did not carry the receptor chimeras was very low. This methodology can in principle be extended to any ligand that acts via clustering of its receptors. PMID- 10670655 TI - Resistance to Paracoccidioides brasiliensis infection is linked to a preferential Th1 immune response, whereas susceptibility is associated with absence of IFN gamma production. AB - The secretion of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-4, IL-5, and IL-10 by antigen-stimulated lymph node cells, eosinophil maturation, and the antibody isotypes produced were examined during intraperitoneal infection of susceptible (B10.A) and resistant (A/Sn) mice with Paracoccidioides brasiliensis. Lymph node cells from resistant mice produced early and sustained levels of IFN gamma and IL-2, whereas susceptible animals secreted low to undetectable amounts of these type 1 cytokines. Both mouse strains presented late and transient production of IL-4, whereas IL-10 was produced constantly throughout the course of disease. Resistant animals produced increasing levels of IL-5 in the chronic phase of the infection (from the eighth week on), whereas susceptible mice showed two peaks of IL-5 production, at the first and twelfth weeks after infection. Only the susceptible strain presented medullary and splenic eosinophilia concomitant with the raised IL-5 production. In resistant mice, the levels of IgG2a antibodies were significantly higher than those observed in susceptible mice, which preferentially secreted IgG2b and IgA isotypes. Taken together, these results demonstrate that a sustained production of IFN-gamma and IL-2 and a predominant secretion of IgG2a antibodies are associated with resistance to P. brasiliensis. In contrast, the production of low levels of IFN-gamma, early secretion of high levels of IL-5 and IL-10, eosinophilia, and a preferential secretion of IgG2b and IgA isotypes characterize the progressive disease in susceptible animals. PMID- 10670656 TI - Leptin expression is reduced with acute endotoxemia in the pig: correlation with glucose, insulin, and insulin-like growth factor-1 (IGF-1). AB - Leptin has been implicated in the regulation of anorexia associated with cachexia in rodents and humans. Regulation of leptin expression is under complex endocrine and metabolic control. To determine if leptin expression is regulated by acute inflammation and to define the endocrine and metabolic factor(s) that regulates leptin expression during acute inflammation, castrate male pigs (ad libitum fed, used as their own controls) were treated with saline (control period) and endotoxin (lipopolysaccharide [LPS] period). Frequent blood samples were collected to identify dynamic changes in hormones and metabolites that are known to regulate leptin expression. LPS caused fever and elevated plasma cortisol (p < 0.0004), tumor necrosis factor-alpha (TNF-alpha) (p < 0.0001), and plasma nonesterified fatty acids (NEFA) (p < 0.001) compared with control. Circulating insulin (p < 0.01), glucose (p < 0.003), and insulin-like growth factor-1 (IGF-1) (p < 0.0001), as well as adipose leptin mRNA abundance (p < 0.01), were profoundly reduced following LPS treatment compared with control. Our data indicate that during acute endotoxemia (1-10 h after injection), leptin gene expression is decreased compared with ad libitum fed animals and is more closely related to energy homeostasis than cytokine profiles in plasma. PMID- 10670657 TI - IFN-alpha1 plasmid construct affords protection against HSV-1 infection in transfected L929 fibroblasts. AB - The purpose of the present study was to evaluate the resistance against herpes simplex virus type 1 (HSV-1) using an interferon-alpha1 (IFN-alpha1) transgene in specifically targeted cells in vitro. Transfection of mouse fibroblast L929 cells with an IFN-alpha1 plasmid construct reduced viral load and viral gene expression in a time-dependent fashion. Supernatants from IFN-alpha1-transfected cells augmented natural killer (NK) cell activity, and such an effect was antagonized with neutralizing antibody to IFN-alpha/beta. In addition, transfected cells displayed an increase in the IFN inducible genes (2',5'-oligoadenylate synthetase [2',5'-OAS], T cell-specific guanine nucleotide triphosphate-binding protein, IFN regulatory factor 1 [IRF-1], and major histocompatibility complex [MHC] class I) compared with plasmid vector-treated controls. Collectively, these results show that IFN-alpha1 transfection of cells in vitro induces or upregulates a spectrum of IFN-regulated genes involved in the direct or indirect antiviral action of this cytokine. In addition, the transgene significantly increases the resistance of transfected cells in vitro to HSV-1 infection. PMID- 10670658 TI - Models for predicting subjective quality of life in individuals with traumatic brain injury. AB - The objective of this study was to compare the utility of ICIDH-based models and needs-based models for predicting subjective quality of life in individuals with traumatic brain injury (TBI). Using an existing data base of individuals with TBI living in the community, seven predictive models were tested using multiple regression analyses. In comparing adjusted R2 associated with each of seven models, it was concluded that needs-based models using subjective indicators clearly predict more variance in measures of life satisfaction, or subjective well-being, than do either type of model relying on objective measures. It is suggested that, in documenting 'outcomes' of rehabilitation, the degree to which the focal individual's important needs are met defines more directly his/her well being than do measures of impairment, disability or handicap. PMID- 10670659 TI - The sensitivity and specificity of self-reported symptoms in individuals with traumatic brain injury. AB - In this study, self-reported symptoms (cognitive, physical, behavioural/affective) from the TIRR Symptom Checklist are compared across six panels: 135 individuals with mild TBI, 275 with moderate/severe TBI, 287 with no disability, 104 with spinal cord injury, 197 who are HIV positive and 107 who had undergone liver transplantation. Participants with TBI and SCI were at least 1 year post-injury. Individuals with TBI reported significantly more symptoms than other panels. Symptom reports in the TBI panels were independent of demographic variables (gender, education, income, ethnicity, age), as well as time since injury and depression. Five of the 67 symptoms were found to be sensitive/specific to TBI in general; 25 symptoms were sensitive/specific to mild TBI (23 were cognitive, one physical and one behavioural/affective). Implications of these results in terms of current debates about the 'reality' of symptom reports in individuals with mild TBI are discussed, as well as implications for using symptom checklists for TBI screening. PMID- 10670660 TI - The unveiling of traumatic brain injury in an HIV/AIDS population. AB - This study examines the frequency of traumatic brain injury (TBI) in an HIV/AIDS population and its associated symptomatology. A panel of 173 individuals with HIV were split into two groups--those who have experienced a blow to the head within their lifetime (n = 128) and those who have not (n = 45). Self-reported symptoms from the TIRR Symptom Checklist were compared across both HIV panels, individuals who identified as traumatically brain injured (n = 416), and individuals with no disability (n = 282). Six clusters of symptoms (total, cognitive, physical, affective/behavioural, five symptoms sensitive and specific to TBI in general and 25 symptoms sensitive and specific to mild TBI) were analysed in a MANOVA, controlling for the demographic variables that were correlated with total symptoms, including panel membership, education, annual household income and substance use history. Significant main effects were found for panel membership. Individuals with HIV and a history of blow to the head reported a higher number of total symptoms and the 25 symptoms specific to mild TBI. The significance of these findings acknowledges the need to recognize the frequency of TBI in an HIV population and the subsequent need to provide the appropriate interventions that will lead to an enhanced overall quality of life. PMID- 10670661 TI - Axis II psychopathology in individuals with traumatic brain injury. AB - PRIMARY OBJECTIVES: To determine the frequency and nature of post-TBI personality disorders (PDs) in a community-based sample of individuals with TBI. RESEARCH DESIGN: One hundred individuals with TBI were administered a structural clinical interview to determine Axis II psychopathology. METHODS OF PROCEDURES: The Structured Clinical Interview for DSM-IV Personality Disorders, Clinician Version (SCID II) was used to determine 12 Axis II personality disorders. SCID II questions were modified so that symptom onset could be rated as occurring pre injury vs. post-TBI. Data were analysed using student T-tests, chi-square analysis and one way analyses of variance. OUTCOMES AND RESULTS: Pre-TBI PDs were diagnosed in 24% of the sample; antisocial PD and obsessive-compulsive PD were the most common diagnoses. Post-TBI, 66% of the sample met criteria for at least one PD, with PDs independent of TBI severity, age at injury, and time since injury. The most common post-TBI PDs were: borderline, avoidant, paranoid, obsessive-compulsive and narcissistic. Men were more likely to be diagnosed with antisocial PD and narcissistic PD. Individuals with pre-TBI PDs were at greater risk of acquiring additional psychopathology post-TBI. Personality traits endorsed by more than 30% of the sample post-TBI reflected loss of self confidence, attempts to cope with cognitive and interpersonal failures and negative affect. CONCLUSION: These findings argue against a specific TBI personality syndrome, but rather a diversity of personality disorders reflective of the persistent challenges and compensatory coping strategies developed by individuals post-TBI. Prospective need for clinical assessment, pro-active education and focused treatment approaches are discussed. PMID- 10670662 TI - Predictive validity of the Neurobehavioural Cognitive Status Examination (NCSE) in a post-acute rehabilitation setting. AB - Within the context of a post-acute rehabilitation setting, association and agreement between results from the Neurobehavioural Cognitive Status Examination (NCSE) and from the neuropsychological (NP) evaluation are examined. All participants (n = 48) had sustained a severe traumatic brain injury and NCSE testing preceded NP testing by an average of 1 month. A significant relationship and fair classification agreement (i.e. presence or absence of cognitive impairment) was found between the overall results from NCSE and NP evaluation. Significant relationships were also observed between most NCSE subtests and paired NP tests thought to be assessing the same cognitive domains. However, the classification agreement (i.e. the presence or absence of deficient performance) between most NCSE subtests and paired NP tests was poor. The findings are discussed from the standpoint of individual treatment planning. PMID- 10670663 TI - Correlates of depression in adults with severe traumatic brain injury and their carers. AB - Although it is generally accepted that social support plays a role in the maintenance of psychological well-being, there has been relatively little direct investigation of the role that social support may play in affecting post-injury depressive symptoms and mediating the effects of traumatic brain injury (TBI). Consequently, social support was selected as the framework within which to investigate possible indicators of depression in adults with severe TBI and their carers. The authors were interested in the degree of association between social support and the criterion variable of depression in the context of demographic and disability-related variables that have been identified as significant correlates of depression. Thirty-five adults with severe TBI (PTA > 7 days) and their primary carers participated in the study. Time post-injury ranged from 3.5 10 years and all the participants were living in the community. Fifty-seven per cent of the adults with TBI and 60% of their carers were classified as showing significant symptoms of depression. As hypothesized, social support contributed significantly to the prediction of depression. In particular, strong-tie support appeared to be an important indicator of well-being for both the participants with TBI and their carers. PMID- 10670664 TI - Interpretation and comprehension of linguistic humour by adolescents with head injury: a group analysis. AB - The aim of the present study was to determine the ability of adolescents with a head injury to interpret and comprehend linguistic humour. Nine adolescents with head injury aged between 12 years 1 month and 15 years 4 months, and nine individually matched adolescents aged between 12 years 1 month and 16 years 1 month were administered a humour test, a standard language battery, the CELF-3, and the Self-Esteem Index. The test of humour abilities required each subject to recognize and select an explanation from a group of three, as to what made each item funny. Items were based on morphological, semantic and syntactic humour elements. Comparison at a group level demonstrated that adolescents with head injury performed significantly poorer in the interpretation and comprehension of linguistic humour than a group of individually matched peers. Contrary to expectations, a relationship between the level of self-esteem and humour comprehension did not exist. The findings of the present study suggest that further research into the effects of head injury on linguistic humour in adolescents is warranted, particularly from a case-by-case perspective. PMID- 10670665 TI - Feather pecking in groups of growing bantams in relation to floor litter substrate and plumage colour. AB - 1. As part of a programme investigating the causation of pecking damage in fowls, this experiment tested a proposal that birds may receive more feather pecks when their plumage colour contrasts with floor litter colour, because litter particles on plumage (as a consequence of dustbathing) may then have greater stimulus value. 2. Groups consisting of 7 light- and 7 dark-coloured bantams were reared from 1 to 11 weeks of age in pens with either wood shavings (light coloured, n=6) or peat (dark coloured, n=6) floor litter. 3. Feather loss from pecking commenced in the 3rd week of life and increased thereafter, but observed pecking damage scores were not consistent with the hypothesis being tested. 4. Despite many more pecks at birds being seen (over 10 weeks) in the wood shavings groups' (661 at particles on plumage, 1795 not at particles) than in peat groups (205, 787), there was no effect of litter substrate on pecking damage. The only evidence supporting the proposal was the finding that, in groups on wood shavings, significantly more pecks at particles on plumage were directed from light coloured birds towards dark ones, than from light to light, dark to light, or dark to dark. 5. Feather eating was confirmed from the presence of feather material in 2% to 15% of faecal droppings collected from each group at 11 weeks, but these proportions were not correlated with pecking damage scores. 6. The results imply that only some feather pecks/pulls were damaging and only some eaten feathers were pulled from other birds. PMID- 10670666 TI - Day length affects feeding behaviour and food intake in adult male emus (Dromaius novaehollandiae). AB - 1. In south-western Australia, male and female emus decrease their food intake when they start breeding in early winter and increase their intake during spring and summer when the breeding season and egg incubation are finished. 2. This annual feeding cycle seems to be under the influence of several environmental factors. Here, we tested the importance of photoperiod using male emus kept in light-controlled rooms with ad libitum access to food and water. 3. Long days increased food intake whereas short days decreased it. Emus fed only during the light hours. 4. Frequency of meals was similar under the 2-day lengths but meal duration was shorter when the emus were on short days than when they were on long days. Thus, day length seemed to affect appetite but not interest in food. 5. Further investigations are needed to test whether these changes in feeding behaviour are a direct consequence of day length or if they are secondary to photoperiod-driven changes in sexual activity. PMID- 10670667 TI - Incidence of pecking damage in growing bantams in relation to food form, group size, stocking density, dietary tryptophan concentration and dietary protein source. AB - 1. This paper reports 4 experiments with groups of 10 to 20 growing bantams in multi-unit brooders, which investigated effects of certain environmental and dietary factors on development of feather pecking damage to 6 weeks of age. Damage was assessed according to a subjective scoring system. 2. A test of food form (pellets, mash, mash diluted with 100 g/kg powdered cellulose) confirmed that pecking damage tends to be greater with pellets than with mash but there was no significant difference between the low damage scores associated with undiluted and diluted mash treatments. 3. A test of group size (10, 20 birds) and stocking density (744, 372, 186 cm2/bird) showed that variation in pecking damage was associated with group size x density interactions. 4. A test of dietary supplementation with L-tryptophan (0, 10, 20 g/kg) showed suppression of pecking damage with the higher (20 g/kg) dose, compared with the control (0 g/kg) treatment. 5. A test of dietary protein source (plant, mainly animal, mainly semipurified) showed no difference in pecking damage scores between treatments. PMID- 10670668 TI - Stress-induced oviposition delays in laying hens: duration and consequences for eggshell quality. AB - 1. This study investigated relationships between the timing of stress, duration of oviposition delays and consequences for eggshell quality, in 2 experiments with ISA Brown hens. 2. Periods of up to 6 h of environmental stress (relocation from an individual cage to a larger one containing 3 unfamiliar hens), commencing up to 4.5 h before predicted oviposition time, reliably induced oviposition delays. Many hens still retained their egg when stress ended. 3. Ovipositions that occurred during stress were never delayed for more than 3.0 h beyond the expected time. Ovipositions that occurred after the period of stress ended usually did so after <1 h if the delay at the end of stress was less than about 2.4 h. Delays that ended either during stress or <2 h after stress ended were classified as short-term. 4. If an oviposition delay was more than about 2.4 h when stress ended, the egg concerned was usually laid much later, after a delay of 7 to 15 h. Delays that ended >5 h after stress ended were classified as long term. 5. Eggs delayed long-term were white-banded and the subsequent egg was slab sided, or occasionally soft-shelled. Short-term delays often caused eggshell dusting with varying amounts of superficial calcification. Hence, duration of oviposition delay affects both the number of abnormal eggshells and the degree of abnormality. 6. The present findings are of potential importance to both the egg industry and breeders, because abnormal eggshells cause downgrading and can impair embryonic development. Also, numbers of abnormal eggshells and degree of abnormality can be used as indicators of environmental stress. PMID- 10670669 TI - Effects of environmental enrichment, fluorescent and intermittent lighting on injurious pecking amongst male turkey poults. AB - 1. Under commercial and experimental conditions domestic turkeys often cause injuries to pen-mates by repeated pecking, sometimes fatally. Environmental enrichment or lighting manipulations might be used to mitigate such injurious pecking. 2. This study examined responses to 4 treatments (2 rooms/treatment) of 8 groups of 100, non-beak trimmed, non-desnooded, male domestic turkeys from 1 to 35 d of age. 3. Birds of 1 treatment were reared under conditions approximating to commercial rearing (12L:12D incandescent, Control) whereas the experimental treatments were 12L:12D incandescent plus supplemental ultraviolet radiation, straw supplementation of litter, pecking substrates and visual barriers (Enriched), 12L:12D fluorescent lighting (Fluorescent), and 2(2L:3D):2L:12D incandescent (Intermittent). 4. Compared to control birds, the incidence of injuries caused by wing or tail pecking were both lower in the Enriched but not significantly different in the Fluorescent or Intermittent. 5. Injuries caused by head pecking did not occur in the Enriched rooms but were observed in at least 1 of the rooms with Control, Fluorescent and Intermittent treatments. 6. Despite considerable environmental differences between treatments, there was remarkable consistency within each type of injurious pecking in age at which injuries were 1st recorded (wing pecking, 9.38+/-1.31 d; tail pecking, 20.43+/-2.42 d; head pecking, 27.8+/-2.13 d). The roles of feather emergence, hierarchy formation in wild turkey poults and appearance of feathers are discussed as possible explanations of these consistencies. PMID- 10670670 TI - Analysis of risk factors for the occurrence of feather pecking in laying hen growers. AB - 1. Potential risk factors for the occurrence of feather pecking in laying hen growers raised under commercial conditions were investigated on Swiss farms with more than 500 rearing places. On-farm interviews were conducted on a sample of 64 flocks which represented 42.6% of all farms concerned. 2. All variables considered were dichotomised and their univariate correlation with the occurrence of feather pecking was tested for significance at P <0.20 using chi2 tests. Logistic regression with backward elimination was then used with the significant variables to identify the potentially most important factors influencing feather pecking. These variables included stocking density, light intensity, intensity of care, access to elevated perches, access to a roofed and littered outdoor area ('bad weather run'), time of access to the feeding facilities of the housing system, stocking density in the restricted area at the beginning of the rearing period, additional open feeding areas in the beginning and air quality. 3. The final model contained stocking density and access to elevated perches as significant factors (P<0.05). Flocks kept in high density (> or = 10 birds per m2) and with no access to elevated perches were 6.4 (95% Confidence interval 1.7 to 24.2) and 4.0 (95% Confidence interval 1.2 to 12.9) times more likely to be affected by feather pecking, respectively. 4. The study identified 2 risk factors for the occurrence of feather pecking in flocks of laying hen growers reared under commercial conditions. It is concluded that in order to reduce feather pecking chicks should be reared at low density and with access to elevated perches. PMID- 10670671 TI - Heritabilities and genetic correlations of body weights and feather length in growing Muscovy selected in Taiwan. AB - 1. Heritabilities and genetic correlations in the base population of a closed strain of Muscovy duck, moderately selected for body weight at 10 weeks of age, have been estimated from the data of 9 successive generations for the following traits: male and female body weight at 10 and 18 weeks of age (BW10m, BW18m, BW10f, BW18f) and length of the 8th primary feather at 10 weeks of age (F110m, F110f). 2. Multivariate REML with an animal model was used, pooling data from the 9 generations (3283 and 3289 male and female offspring respectively). The same trait expressed in male and female was considered as 2 different traits. 3. The 8th primary feather was longer in females than in males by 6% to 22% at 10 weeks of age. Body weight was heavier in males than in females by 42% to 58% at 10 weeks of age and by 57% to 75% at 18 weeks of age. 3. The heritability estimates for body weight traits showed moderate values, being a little higher for females than for males at the same age, increasing with age from h2=0.24 at BW10m to h2=0.43 at BW18f. 4. The heritability estimates for feather length showed that a greater response would be obtained in selection for male feather length (h2=0.37) than for female length (h2=0.14). Both have high genetic correlations with body weight so they could be indirectly improved. 5. Heritabilities of the difference in body weights between males and females at 10 weeks (h2=0.07) and 18 weeks of age (h2=0.10) were small, as well as for feather length (h2=0.10). It would probably be difficult to modify sexual dimorphism in body weight through selection. 6. Genetic correlations between BW10m, BW18m and BW10f, BW18f were respectively r(g)=0.77 and r(g)=0.80. They were larger for body weight at the same ages between males and females, r(g)=0.90 (r(g)=0.88 between F110m and F110f). Body weight in males and females at the same age should be better considered as 2 different traits in a selection programme. 7. The cumulated predicted genetic gains expressed per unity of the genetic standard deviation (sigma(g)) over the 8 generations of selection were 1.3 sigma(g) and 1.4 sigma(g) respectively for the BW10m and BW10f. The predicted correlated responses were 1.2 sigma(g) for body weights at 18 weeks of age, 0.9 sigma(g) and 0.7 sigma(g) for F110f and F110m respectively. PMID- 10670672 TI - Haematological and immunological variables in a domesticated and wild subspecies of ostrich (Struthio camelus). AB - 1. Domesticated ostriches have been selected rigorously for productive traits with little concern for immunological responses, in contrast to wild ostriches. 2. We hypothesised that the immunological responses of wild and domesticated ostriches would differ. Total leucocyte counts, differential counts, heterophil: lymphocyte ratios, phagocytic activity, lysosome levels and anti-sheep red blood cell (SRBC) antibody titres (total, IgG, IgM) were compared between domesticated (n=3) and wild (n=3) ostrich subspecies. 3. Total leucocytes, lymphocytes and heterophils were similar in the 2 subspecies, but basophils and eosinophils were lower in the wild than in the domesticated ostriches. Lysosome concentrations and phagocytic activities were higher in the wild ostriches. 4. Total and IgM antibody titres to SRBC reached peak values quicker in the domesticated than in wild ostriches. IgG development patterns were similar. 5. The results suggest that a stronger non-specific immune response was shown by the wild ostriches (higher phagocytosis and lysozymes) whereas a stronger specific immune response was shown by the domesticated ostriches (peak values of anti-SRBC antibody titres were reached more quickly). PMID- 10670673 TI - Efficacy of a commercial competitive exclusion product against Campylobacter jejuni. AB - 1. Newly-hatched broiler chicks were treated orally with a commercial competitive exclusion product (Broilact) in 3 replicate trials 2. After 24 h the treated chicks and untreated control chicks were challenged orally with approximately 10(4) cfu of Campylobacter jejuni. 3. The caeca of the birds were examined quantitatively for campylobacter 12 d after the challenge. 4. In 3 separate trials, the treatment prevented or reduced colonisation of the challenge organisms in the caeca. The percentage of colonised birds varied from 0% to 62% in the treated groups and was 100% in the control groups. The average number of campylobacter was considerably lower in the treated groups than in the control groups. PMID- 10670674 TI - Tenderness, moisture loss and post-mortem metabolism of broiler Pectoralis muscle from electrically stimulated and air chilled carcases. AB - 1. This study was to evaluate the effects of post-mortem electrical stimulation (ES) on carcase moisture loss and on tenderness, R-value and pH of breast fillets following air chilling. 2. In each of 4 replications, 8 birds were electrically stimulated and 12 birds were controls. The ES birds were stimulated at the head in a 1% saline bath (450 V, 0.45 A, 2 s on/1 s off for 7 pulses). After evisceration the carcases were air chilled in a cooler at 1 to 2 degrees C with an average relative humidity of 91% and an air speed of 44 m/min. 3. Breast fillets were harvested at 2 and 4 h postmortem from both ES and control carcases and also at 8 h postmortem from control carcases to determine moisture loss pH and R-value. 4. Although there was no significant effect of ES on shear value at 2 h postmortem, the ES fillets had a lower shear value mean than the control fillets at 4 h postmortem. There was no significant difference between the shear value means of the ES 4 h fillets and the control 8 h fillets. 5. ES accelerated the normal post-mortem decline in pH at both 2 and 4 h postmortem. 6. The R-value means for the control and ES samples were similar 2 h but the R-value mean of the ES samples was greater than the control at 4 h postmortem. 7. The results suggest that, when followed by air chilling, ES accelerates postmortem metabolism, reduces ageing by up to 50%, and has no effect upon evaporative moisture loss. PMID- 10670675 TI - Transmission electron microscopy of the vertical crystal layer and cuticle of the eggshell of the domestic fowl. AB - 1. Eggshell pieces (1 cm2) were decalcified using a solution of EDTA (200 g/litre, pH 6.9 to 7.0) in paraformaldehyde (2 g/litre) and glutaraldehyde (0.5 g/litre) in phosphate buffer. 2. They were prepared for transmission electron microscopy (TEM). 3. TEM identified a vertically aligned matrix associated with the vertical crystal layer (VCL). It is hypothesised that the vertical orientation of calcite crystals in the VCL is closely linked to this vertical matrix. 4. TEM also revealed the presence of a 2-layered cuticle, the inner layer containing vesicles which were absent in the outer. 5. Cuticular vesicles contain hydroxyapatite and are thought to play a role in the termination of shell formation. The current paper presents data relating to microbial apatitic systems that strengthen this hypothesis. PMID- 10670676 TI - Features of eggshell formation in guinea fowl: kinetics of shell deposition, uterine protein secretion and uterine histology. AB - 1. Rate of calcium carbonate deposition, duration of eggshell formation, organic composition of the uterine fluid, morphology of the egg shells and histochemistry of the uterus were studied in guinea fowl to analyse the origin of such thick, strong egg shells. 2. The egg shell was linearly deposited from 6.4 h to 21.8 h after the oviposition of the previous egg. The rate of egg shell deposition was similar to that in laying hens. However, the duration of linear shell deposition was increased by 2.1 h relative to that in hens. This explained the increased egg shell weight observed in the guinea fowl. 3. Intervals between oviposition of intra-clutch eggs were 24 h throughout the laying period. Ovulation occurred just after oviposition of the previous egg in the guinea fowl, as previously observed in hens but the duration of egg white protein deposition, of plumping and of initiation of shell mineralisation were all 1.5 h shorter than in domestic hen. 4. Uterine fluid can only be collected during the growth and terminal phase of shell formation. The electrophoretic profiles of the uterine fluid differed between phases and were somewhat different from those previously observed in the hen. Ovalbumin and ovocleidin-17 were both present in the uterine fluid and also in egg shell extract. Ovocleidin-17 was predominant during the growth phase. 5. The histology of the uterus differed slightly in guinea fowl compared to hens. Ovocleidin and ovalbumin are both secreted by the tubular glands. 6. Examination of radial ultrathin sections of eggshell showed, above the mammillary layer, intricate interlacing of adjacent exospherite in guinea fowl in contrast to the continuous columnar microstructure in hens. 7. The kinetics of egg shell deposition largely explains the increased egg shell weight of guinea fowl. The organic matrix proteins may be associated with the contrast between the structural organisation of the guinea fowl egg shell and that of the hen egg shell. PMID- 10670677 TI - Composition of vitamin supplements in Spanish poultry diets. AB - 1. The composition of 106 vitamin supplements used in about 85% of the Spanish poultry production diets were studied. Vitamin supplements were grouped by production classes and, for broilers and pullets, also by feeding periods. 2. Four vitamins (niacin, alpha-tocopherol, pantothenic acid, and riboflavin) comprised over 87% of the vitamin supplements by weight (choline excluded), whereas alpha-tocopherol and retinol represented from 51% to 60% of the total vitamin cost. 3. The highest and lowest vitamin supplementation rates were for broilers in the starter and withdrawal periods (106 and 44 mg/kg, respectively) and the mean values for breeders, pullets and layers were 104, 58, and 48 mg/kg, respectively. 4. Supplements with higher vitamin contents showed less variability in their composition. Retinol, cholecalciferol, riboflavin and pantothenic acid showed the lowest variability within supplements (6% to 36% CV), whereas alpha tocopherol, menadione, thiamin and biotin showed the highest (40% to 224% CV). 5. Vitamin supplementation rates were compared with requirements, taking into account the dietary contribution. In general, vitamin fortification exceeded the NRC recommendations, using a high safety margin for some vitamins such as vitamin A (from 2.6 to 7.8) and for some poultry classes such as breeders (3.2). PMID- 10670678 TI - Energy utilisation of carbohydrate, fat and protein sources in newly hatched broiler chicks. AB - 1. TME, TMEn and metabolisability (TME/gross energy) of energy-yielding foodstuffs were determined in 1, 3 and 10 d old broiler chicks, using the assay method developed by Murakami et al. (1994), in order to characterise energy utilisation during the 10 d after hatching. 2. TME, TMEn and metabolisability of dextrin and starch were low in chicks aged 1 d, and increased with age up to 10 d. Energy values of glucose and maltose at days 1 and 3 could not be determined because of the sudden death of birds soon after the feeding. TMEn and metabolisability of cereal sources were lower in d-old chicks than in those aged 3 and 10 d. At all ages, maize was better utilised than wheat and sorghum. 3. In the fat sources (coconut oil, beef tallow and safflower oil) no age dependency was observed in TME, TMEn and metabolisability. 4. Bioavailability of soyabean meal and fish meal was lower at 1 d than at 3 d and 10 d. Energy utilisation from casein was the highest among the protein sources tested and it was not age dependent. 5. It is concluded that energy utilisation of carbohydrate and protein sources during 10 d post-hatch tended to increase with age. Among the energy yielding foodstuffs fat sources seem to be better utilised, with no age dependency. PMID- 10670679 TI - Response of broiler chickens to microbial phytase supplementation as influenced by dietary phytic acid and non-phytate phosphorus contents. I. Effects on bird performance and toe ash. AB - 1. Seven-day old male broilers (n=900) were fed on wheat-sorghum-soyabean meal based diets containing 3 concentrations of phytic acid (10.4, 13.2 and 15.7 g/kg; equivalent to 2.9, 3.7 and 4.4 g/kg phytate phosphorus), 2 of non-phytate phosphorus (2.3 and 4.5 g/kg) and 3 of microbial phytase (Natuphos 5000 L; 0, 400 and 800 FTU/kg) in a 19-d trial. The dietary phytic acid contents were manipulated by the inclusion of rice pollard. 2. Each dietary treatment was fed to 5 pens (10 birds/pen) from 7 to 25 d of age. Records of body weight, food intake and mortality were maintained. On d 25, all surviving birds were killed and toe samples were obtained for toe ash measurements. 3. Increasing dietary phytic acid negatively influenced body weight gain, food intake and food/gain. These adverse effects were partially overcome by the addition of microbial phytase. 4. Supplemental phytase caused improvements in weight gain and food efficiency of broilers but the magnitude of the responses was greater in low non phytate phosphorus diets, resulting in significant non-phytate phosphorus x phytase interactions. 5. Toe ash contents were improved by phytase addition but the response was greater at the highest concentration of phytic acid, resulting in a significant phytic acid x phytase interaction. Responses were also greater in low non-phytate phosphorus diets as indicated by significant non-phytate phosphorus x phytase interaction. 6. In general, there was very little difference in the responses to phytase additions at 400 and 800 FTU/kg. 7. The performance responses to added phytase in birds receiving adequate non-phytate phosphorus diets provide evidence for the influence of the enzyme on animal performance independent of its effect on phosphorus availability. PMID- 10670680 TI - Optimum inclusion of field peas, faba beans, chick peas and sweet lupins in poultry diets. I. Chemical composition and layer experiments. AB - 1. Experiments were undertaken to determine the chemical composition and apparent metabolisable energy (AME) of field peas, faba beans, sweet lupins and chick peas and the production of hens when each was included in nutritionally similar diets at 250 g/kg in 2 experiments. 2. Amino acid composition, crude protein and AME agreed well with previously published measurements. Detailed analysis of the non starch polysaccharides (NSPs) showed that sweet lupins were much higher than the other grain legumes in the soluble NSPs and that the NSPs were particularly high in arabinose. The condensed tannin content was highest in field peas followed by faba beans. 3. In the 1st layer experiment over 40 weeks, hen-day egg production was lowest on the faba bean-based diet and egg weight and egg mass were also lowest. Relative viscosity of digesta in the small intestine of hens fed on the sweet lupin-based diet was highest, followed by that of hens fed on field peas. Enlargement of the pancreas was observed in hens consuming chick peas. 4. In experiment 2, in which only sweet lupins and faba beans were used, steam or cold pelleting showed few effects, nor did dehulling of faba beans but egg weight was lower when diets were steam pelleted. Daily food intake was 5.7 g/bird lower on the steam than cold pelleted diets and food conversion ratio tended to be improved (P=0.082). 5. It was concluded that field peas could support good production at 250 g/kg of layer diet. Although chick peas and sweet lupins supported good performance, there was concern about the increased weight of the pancreas and high gut viscosity respectively. Faba beans showed similar hen-d egg production in the 2nd experiment to that of sweet lupins but egg weight tended to be about 0.8 g lower than when on the sweet lupin-based diets. PMID- 10670681 TI - Optimum inclusion of field peas, faba beans, chick peas and sweet lupins in poultry diets. II. Broiler experiments. AB - 1. Three experiments were undertaken to determine the optimum inclusion rates of field peas, faba beans, chick peas and sweet lupins in broiler starter and finisher diets in amounts up to 360 g/kg. 2. In experiment A chickens in cages grown to 21 d on diets with field peas and faba beans gave better growth rate and feed efficiency than those with sweet lupins and chick peas. Growth rate and Food conversion ratio (FCR) improved with increasing amounts of faba beans in the diet while for chick peas growth rate and FCR declined. Digesta viscosity and excreta stickiness scores were much higher on diets with sweet lupins. Steam pelleting improved growth rate and FCR on all diets. 3. In experiment B birds were in cages and grown from 21 to 42 d. There were no differences between grain legumes (when combined for all inclusions) for growth rate, food intake or FCR. Viscosity was again much higher on the sweet lupin-based diets while the pancreas was significantly enlarged on the diets with chick peas, as observed previously in chickens grown to 21 d. Steam pelleting of diets gave a consistent and positive response for weight gain and FCR. 4. Experiment C was carried out in pens each holding 60 birds under semi-commercial conditions and grown to 42 d on starter and finisher diets with the same grain legumes as used previously but each at 2 rates of inclusion similar to those in commercial practice. Field peas at 200 to 300 g/kg and chick peas at 150 to 220 g/kg gave inferior growth to faba beans (150 to 180 g/kg) and sweet lupins (120 g/kg). 5. The results of these experiments allowed tentative recommendations to be made to industry for inclusion rates of these cultivars of the 4 grain legumes. These were: field peas 300 g/kg; faba beans 200 g/kg, chick peas 100 g/kg and sweet lupins <100 g/kg. Wet droppings and high gut viscosity were serious problems with sweet lupins although these were not so obvious in experiment C. PMID- 10670682 TI - Influence of dietary energy, supplemental fat and linoleic acid concentration on performance of laying hens at two ages. AB - 1. The aim of this study was to investigate the influence of metabolisable energy (ME), supplemental fat (SFAT) and linoleic acid (LIN) content of the diet on the productive performance and weight of eggs and egg components of Isabrown hens of 22 or 74 weeks of age. 2. Six diets were formulated to contain the following concentrations of ME (MJ/kg), SFAT (g/kg) and LIN (g/kg), respectively: A) 11.8, 0 and 11.5; B) 11.8, 40 and 11.5; C) 11.8, 40 and 16.5; D) 11.2, 40 and 16.5; E) 11.2, 40 and 11.5; and F) 11.2, 0 and 11.5. Data were collected for 28 d and analysed using linear contrasts to test the effect of SFAT, LIN, ME and their interactions. 3. When the LIN content of the diets was maintained constant at 11.5 g/kg, an increase in the SFAT from 0 to 40 g/kg increased egg weight (63.8 vs 64.5 g; P<0.05), food intake (119 vs 124 g; P<0.01) and energy intake (1.36 vs. 1.42 MJ/d; P<0.01) and body weight change of the hens (-85 vs. 27 g; P<0.001). Supplemental fat also increased yolk (15.8 vs. 16.3 g; P<0.001) and albumen weight (40.8 vs. 42.3 g; P<0.01) but yolk to albumen ratio was not modified. 4. Egg and albumen weights were improved by SFAT in early but not in late producing hens. As a result, yolk to albumen ratio decreased in the younger hens, from 0.371 to 0.357, but increased in the older hens, from 0.408 to 0.415; P<0.01) with fat addition. 5. An increase in the LIN content of the diets from 11.5 to 16.5 g/kg did not modify any of the traits studied. 6. It was concluded that the LIN requirement of the hens for maximal productivity and weight of eggs is 11.5 g/kg or less. Supplemental fat increased the weight of eggs and albumen in the younger but not in older hens and the beneficial effect was independent of its LIN content. PMID- 10670683 TI - Effect of dietary methionine on performance, carcase characteristics and breast meat composition of heterozygous naked neck (Na/na+) birds under spring and summer conditions. AB - 1. Heterozygous naked neck (Na/na+) birds and their normally feathered counterparts (na+/na+) were fed from 0 to 7 weeks on 3 diets differing in methionine concentrations. From 0 to 3 and 3 to 7 weeks, respectively, the concentrations were: low containing 4.3 and 3.3 g/kg; optimum containing 5.0 and 3.8 g/kg and; high with 5.7 and 4.4 g/kg under spring (optimum ambient temperature) and summer conditions (high ambient temperature). Performance, carcase characteristics and breast meat chemical composition were determined. 2. Summer rearing resulted in a decrease in body weight, body weight gain, food consumption, and yields of carcase and breast. The summer temperature effect was more pronounced in males. Under summer temperatures, the protein content of the breast decreased while the fat content increased compared to birds reared in spring. 3. By 7 weeks of age, both genotypes reached similar body weights in the spring experiment while, in summer Na/na+ birds were 3.3% heavier and gained more in the period from 3 to 7 weeks than na+/na+ birds. Carcase and breast yields of Na/na+ birds were greater than in na+/na+ birds. 3. Second order polynomial coefficients of the dietary methionine effect were found to be significant for body weight at 3 and 7 weeks. Daily body weight gain between 3 and 7 weeks was linearly affected by the dietary methionine concentration. There was no interaction between genotype and methionine. 4. Methionine had no significant effect on carcase yield. Second order polynomial coefficients of the dietary methionine effect were found to be significant for breast yield while the methionine effect on abdominal fat was linear. Na/na+ females fed on the low methionine diet had lower protein content than the Na/na+ males. 5. It is concluded that the methionine requirement of Na/na+ birds did not differ from that of their normally feathered counterparts under either spring or summer ambient temperature conditions. PMID- 10670684 TI - Effect of dried tomato pulp on the performance and egg traits of laying hens. AB - 1. Grouped laying hens were fed on experimental diets containing dried tomato pulp (DTP) at inclusion rates up to 120 g/kg and two control diets (A and A+Carophyll). 2. The addition of DTP did not significantly affect egg production, food consumption and efficiency, egg weight and shell thickness. 3. There was a significant improvement in the egg yolk colour in treatments containing DTP. However, this improvement was significantly lower in comparison to the diet which contained carophyll. PMID- 10670685 TI - Intracerebroventricular injection of orexins does not stimulate food intake in neonatal chicks. AB - 1. Recently, 2 novel neuropeptides were discovered, both derived from the same precursor by proteolytic processing, which bind and activate 2 closely related orphan G protein-coupled receptors, Named orexin-A and -B (Sakurai et al., 1998). Both stimulate food intake when administered centrally to rats. 2. Our aim was to elucidate whether central injection of mammalian orexin-A or -B stimulates food intake in the chick. 3. Under conditions of free access to food, orexin-A did not alter the food intake of chicks, but cumulative food intake was significantly suppressed by orexin-B. 4. The orexin-B was then administered to chicks deprived of food for 3 h to confirm its suppressive effect. No significant effect of orexin-B on food intake was detected. 5. Central injection of orexin-B did not modify food intake when appetite was stimulated by fasting. 6. Neither of these orexins appears to stimulate feeding in chicks. PMID- 10670686 TI - Persistent hypoglycemia induced by continuous insulin infusion in broiler chickens. AB - 1. Persistent hypoglycaemia was experimentally induced by insulin infusion to improve understanding of the regulatory mechanisms of blood glucose concentrations specific to chickens. 2. An osmotic minipump containing bovine insulin was implanted to deliver insulin in vivo at a constant rate (11.25 to 45 U/kg BW/d) for 5 d in 4-week-old broiler chickens force-fed a maintenance diet once a d. Birds infused with the highest dose of insulin died within 3 to 4 d. 3. In chickens continuously infused with insulin at 22.5 U/kg BW/d, fasting glucose concentrations in plasma determined every 3 h during the 3rd day of infusion were consistently and significantly lower than in controls. 4. Continuous infusion of insulin at 22.5 U/kg BW/d induced persistent hypoglycaemia (almost one-half the normal blood glucose concentration) lasting for at least 4 d in broiler chickens. 5. Insulin infusion did not significantly change plasma NEFA or protein concentrations and increased plasma GOT activity only at 1 of the daily experimental sampling points. PMID- 10670687 TI - Production and physical characteristics of composted poultry carcases. AB - 1. Experiments were designed to determine whether composting could be a safe and effective method for the disposal of poultry carcases in the UK climate. Laying hen carcases (125) were composted in a wooden compost bin over autumn and winter months, using the United States Department of Agriculture method. 2. The process took 8 weeks and effectively decomposed the carcases to leave only leg and breast bones. The compost was turned once, which ensured that all the material reached the high temperatures (60 degrees to 70 degrees C) required to control pathogens. Salmonella was fully heat-inactivated, indicating that many poultry-associated bacterial pathogens would also have been inactivated. 3. It is concluded that this method is suitable for use in the UK and provides a sanitised fertiliser supplement. PMID- 10670689 TI - Effects of altering dietary fatty acid composition on prostaglandin synthesis and fertility. AB - Several studies over the past 20 years have demonstrated that subjects on diets composed of substances with high levels of n-3 polyunsaturated fatty acids (PUFAs) (e.g. fish) have a decreased incidence of heart disease. On this basis, a recent report from the Department of Health has advised UK consumers to decrease the proportion of saturated as opposed to unsaturated fats in their diet and to increase the ratio of n-3 to n-6 PUFAs. This could be achieved by altering the amounts of these constituents in milk and meat. n-3 Fatty acids can most easily be added to animal feed as either fish oil or linseed oil and can be increased in the blood and milk of ruminants following protection to avoid hydrogenation in the rumen. In western countries the ratio of consumption of n-6 to n-3 PUFAs is greater than 10 and current evidence tends to suggest that a ratio nearer 5 would be more desirable and compatible with cardiovascular well being. As fertility in the UK dairy herd is already poor, it is important to establish whether alterations in dietary n-3 and n-6 PUFAs affects herd fertility before widespread changes in animal diets are recommended. Therefore, this review considers the role played by PUFAs and eicosanoids in fertility, with particular reference to the implications for farm livestock production. The evidence reviewed shows that alteration of the concentration and ratio of n-6 and n-3 PUFAs in feeds can influence prostaglandin synthesis/metabolism in a number of mammalian systems. The changed patterns of prostaglandin synthesis can as a consequence, affect the diverse functions (e.g. hormone secretion) that are normally mediated via prostaglandins. Similarly, changes in prostaglandin synthesis effected through manipulation of PUFAs has a major bearing on fertility (as PGs affect many reproductive parameters, e.g. ovulation). Several studies in cattle and other mammals, show that feeding or infusing different types of fat with varying PUFA content to females can alter: the number and size of ovarian follicles, the ovulation rate, progesterone production by the corpus luteum, the timing of luteolysis and gestational length. In the male most recent work has focussed on sperm production and experiments in fowl have demonstrated clear effects of dietary PUFAs on both the sperm membrane phospholipid composition and on fertilizing ability. PMID- 10670688 TI - Sphingolipids stimulate cell growth via MAP kinase activation in osteoblastic cells. AB - Ceramide, ceramide-1-phosphate (C1P) sphingosine (SPH) and sphingosine-1 phosphate (S1P) effects on proliferation and extracellular-signal regulated kinases, ERKs (also known as MAPKs), activation were investigated in human and rat osteoblastic cells. MAPK activation was sphingolipid-specific in cells from both species. In human osteoblastic cells, S1P and C1P markedly stimulated ERK2 phosphorylation with a slight increase in phosphorylation of ERK1. SPH nor ceramide induced phosphorylation of either ERK isoform. In rat osteoblastic cells, SIP, ceramide and SPH stimulated phosphorylation of both isoforms. C1P did not induce phosphorylation of ERK1 but produced a mild increase in phosphorylation of ERK2. In human cells, only S1P significantly (P<0.05) increased osteoblastic cell proliferation, while in the rat cells all four sphingolipids significantly (P<0.05) induced proliferation. The calcium channel blocker verapamil blocked (P<0.05) these effects in both cell types. The MAPK inhibitor, PD98059, inhibited (P<0.05) the mitogenic effect of SIP in human cells. In rat cells, PD98059 effects were less substantial but significant for S1P and C1P. This study demonstrates that sphingolipids are mitogens for both human and rat osteoblastic cells with the MAPK pathway and calcium mediating in part these effects in a species specific manner. PMID- 10670690 TI - The effects of a diet rich in docosahexaenoic acid on organ and vascular fatty acid composition in spontaneously hypertensive rats. AB - Previous research has shown that dietary docosahexaenoic acid (DHA) attenuates the development of high blood pressure in young spontaneously hypertensive rats (SHR). The purpose of this study was to investigate the effects of dietary DHA on organ and vascular fatty acid composition in SHR. Given the important structural and functional role of fatty acids in cell membranes, alterations in fatty acid composition may contribute to the antihypertensive effect of DHA. SHR were fed a purified diet containing either a corn/soybean oil mixture (CSO, control) or a DHA-enriched oil for 6 weeks. The DHA diet markedly increased the levels of DHA in the aorta, renal artery, plasma, liver, heart, kidney, and lung by 5-, 15-, 7 , 6-, 3.8-, 3.5-, and 8.8-fold (P<0.001), respectively. The levels of eicosapentaenoic acid were also increased while there was a concomitant reduction in arachidonic and adrenic acids. Therefore, dietary DHA increases the incorporation of omega-3 polyunsaturated fatty acids in specific organs and vascular tissue in SHR at the expense of omega-6 polyunsaturated fatty acids. PMID- 10670691 TI - A comparison of the effects of indomethacin and NS-398 (a selective prostaglandin H synthase-2 inhibitor) on implantation in the rat. AB - Indomethacin (25 microM) inhibited the amount of prostaglandin (PG) E2, PGF2alpha and 6-keto-PGF1alpha synthesized by homogenates of day 5 pregnant rat uterus by 80-92%. In contrast, 25 microM NS-398, a selective inhibitor of prostaglandin H synthase-2 (PGHS-2), inhibited the synthesis of these three prostaglandins by homogenates of the same tissue by only 37-60%. Since it has been reported that idomethacin and NS-398 inhibit PGHS-2 with similar potencies and that indomethacin (unlike NS-398) is a potent inhibitor of PGHS-1, it may be concluded that prostaglandin production by homogenates of the rat uterus on day 5 of pregnancy is due to the activities of both PGHS-1 and PGHS-2. The administration of indomethacin (3 mg/kg) twice daily on days 3 and 4 of pregnancy reduced the implantation rate by 77%, whereas the similar administration of NS-398 (6 mg/kg) had no significant effect on implantation. It may be concluded that either the dose of NS-398 used was too low to affect uterine PGHS-2 activity in vivo sufficiently to prevent implantation, or that inhibiting uterine PGHS-2 activity in vivo has no effect on the implantation mechanisms or is compensated for by increased PGHS-1 activity such that the implantation process is not impaired. PMID- 10670692 TI - Effects of prostaglandin E1 on high density lipoprotein-phospholipid composition. AB - Serum high-density phospholipids (HDL-phospholipids) composition was determined in rats treated with prostaglandin E1 (PGE1) and control group treated with isotonic saline. Total phospholipids and HDL-phospholipids levels at serum were found lower in rats treated with PGE1, than in controls. Considering the individual phospholipid classes of HDL, we observed that phosphatidylocholine (PC), phosphatidylinositol (PI) and diphosphatidylglycerol (DPG) serum concentrations were significantly higher in treated rats than in controls (P<0.001, P<0.005 and P<0.05 respectively). Furthermore, the serum concentrations of lysophosphatidylocholine (LPC) and phosphatidylserine (PS) were significantly lower in treated rats than in controls (P<0.01 and P<0.001 respectively). These findings suggest that PGE1 influences the composition of HDL-phospholipids and possibly modifies their action on lipid metabolism. PMID- 10670693 TI - Prostaglandins and cyclic nucleotides in the urinary bladder of a rabbit model of partial bladder outlet obstruction. AB - Bladder outlet obstruction (BOO) is a common disorder that is associated with altered bladder structure and function. For example, it is well established that BOO results in hypertrophy and hyperplasia of the bladder smooth muscle as well as detrusor instability. Since prostaglandins (PGs) and cyclic nucleotides (cyclic AMP [cAMP] and cyclic GMP [cGMP]) mediate both smooth muscle tone and proliferation, it is reasonable to suggest that changes in their levels may be involved in the pathophysiology of BOO-associated bladder disorders. Hence, the objective of this study was to investigate cyclic AMP, cyclic GMP and prostaglandins in the bladder of a rabbit model of BOO. BOO was induced in adult male New Zealand White rabbits. After 3 weeks, urinary bladders were excised, weighed and cut into segments. They were then incubated with stimulators of PGs, cAMP and cGMP and the formation of PGs, cAMP and cGMP were measured using radioimmunoassays. There was a significant increase in the obstructed bladder weights (P=0.002). The formation of PGE2, PGI2, cAMP and cGMP was significantly diminished in the detrusor (P<0.05) and bladder neck (P<0.05) in the BOO bladders compared to age-matched controls. Since PGE2, PGI2, cAMP and cGMP are known to inhibit the proliferation of smooth muscle cells (SMCs), the decreased synthesis of these factors, in BOO, may play a role in bladder SMC hypertrophy/hyperplasia. Our study points to the possible use of drugs that modulate the NO-cGMP and/or PG cAMP axes in BOO-associated bladder pathology. PMID- 10670694 TI - Involvement of cyclooxygenase-2 in proliferation and morphogenesis induced by transforming growth factor alpha in gastric epithelial cells. AB - Transforming growth factor alpha is one of the most potent growth factors for gastrointestinal epithelium. In this study, we examined the roles of cyclooxygenase-2 on proliferation and morphogenesis of RGM1 rat gastric epithelial cells after stimulation with transforming growth factor alpha in vitro, RGM1 cells increased expression of cyclooxygenase-2 messenger RNA 20-60 min after stimulation with transforming growth factor alpha. Transforming growth factor alpha stimulated [3H]thymidine incorporation and tubulogenesis of RGM1 cells in collagen matrix, both of which were significantly suppressed by treatment with a cyclooxygenase-2 specific inhibitor, NS-398 or cyclooxygenase-2 antisense oligonucleotide. Both of the treatment lowered prostanoid production by enzyme immunoassay. The transforming growth factor alpha-induced expression of cyclooxygenase-2 is followed by cell proliferation and development of tubular morphology of RGM1 gastric epithelial cells. Treatment with cyclooxygenase-2 inhibitor and cyclooxygenase-2 antisense oligonucleotide suppressed these responses induced by transforming growth factor alpha suggesting the involvement of cyclooxygenase-2 in proliferation and morphogenesis in gastric mucosal epithelium. PMID- 10670695 TI - Cytokines regulate prostaglandin H synthase-1 transcription in human amnion derived cells. AB - We have isolated the prostaglandin H synthase-1 (PGHS-1) promoter from the human amnion cell line WISH by long template PCR. The fragment was 1124 base pairs in length and shared a 96% sequence identity with the sequence in GenBank. The putative transcription start site is located 18 bp upstream of the start codon. The sequence is TATA-less, but contains multiple Sp-1 sites and a GC box at -132. The fragment was subcloned into the promoterless reporter construct pBLCAT3 to produce the promoter reporter construct pPGHS1CAT. pPGHS1CAT expression in amnion derived AV3 cells was inhibited by the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). PGHS-1 mRNA levels however, were unchanged over a 16-h time course with either treatment. These results suggest that PGHS-1 transcription is regulated in a negative manner by cytokines in human amnion-derived cells. PMID- 10670696 TI - Cytotoxicity of combined essential fatty acids on a human prostate cancer cell line. AB - In this study the effect of single and concomitantly added n-6 or n-3 polyunsaturated fatty acids (PUFAs) was investigated on human prostate cells. Data obtained from the single fatty acids (FAs) experiments showed that except for oleic acid (OA), arachidonic (AA) and linoleic acid (LA), which had very little (less than 10% cells dead) effect on the cells, an increase in dead cells was observed at physiological concentrations of, eicosapentaenoic acid (EPA), gamma-linolenic acid (GLA) and alpha-linolenic acid (ALA). However, this was not the case when combining these acids at physiological concentrations. A slight increase in cell death was only obtained with three combinations of ALA, namely with AA, OA, or GLA. Other combinations with ALA, such as with LA or EPA, had respectively no effect on cell number or increased the cell number by causing less cells to die. Other PUFAs combinations tested, did not show the three groups mentioned with ALA, but only the last two types, namely, no effect, or a decrease in the amount of cell death. The latter might mean that the FA combination had stimulated the cells, since a decrease in the amount of dead cells was observed. Therefore, it is concluded that the characteristics of combined FAs may differ from single FAs, which may explain some controversies in the literature and in response to treatments. PMID- 10670697 TI - Disease patterns in field and bank vole populations during a cyclic decline in central Finland. AB - Declining field vole (Microtus agrestis) and bank vole (Clethrionomys glareolus) populations were sampled (117 field voles and 34 bank voles) in south-central Finland during the winter of 1988-89. The last surviving field voles were caught in April and bank voles in February. A subsample (16) of the April field voles were taken live to the laboratory for immunosuppression. The histopathology of the main internal organs and the presence of aerobic bacteria and certain parasites were studied. In the lungs, an increase in lymphoid tissue, probably caused by infections, was the most common finding (52% of all individuals). The prevalences in the voles, in the whole material, of Chrysosporium sp. and Pneumocystis carinii in lungs were 13 and 10% in field voles, and 9 and 0% in bank voles, respectively. Cysts of Taenia mustelae (9 and 27%) were the most common pathological changes in the liver. Enteritis was also rather common (14 and 34%). In field voles the prevalences of Frenkelia sp. in the brain and Sarcocystis sp. in leg muscles were low (both 6%). Bordetella bronchiseptica was commonly (31%) isolated from field vole lungs and Listeria monocytogenes from the intestines (34%). Salmonella spp. could not be found. The dynamics and abundance of inflammations in the lungs and intestines, as well as B. bronchiseptica isolations from the lungs, indicate that obvious epidemics took place in declining vole populations. Of the Luhanka subsample of 16 field voles brought to the laboratory in April, one died of listeriosis, two of Bordetella, and five died for unknown reasons. Even if small mustelids are the driving force in microtine cycles, it is possible that diseases also contribute to the decline. PMID- 10670698 TI - Natural killer activity in mice infected with rabies virus and submitted to P. acnes (Propionibacterium acnes) as immunomodulator. AB - The natural killer (NK) activity and lethality were evaluated in swiss mice experimentally infected with street rabies virus and submitted to immunomodulation by P. acnes (formerly Corynebacterium parvum). The infected animals were sacrificed at different times and spleen non-adherent cells were obtained through ficoll-hypaque gradient and depletion of glass-adherent cells. Immunosuppression was observed in rabies virus infected mice correlated with lower NK activity in clinically ill animals. Higher NK activity and percentual of survival were observed in the group submitted to P. acnes. The increased survival correlated with higher NK activity induced by P. acnes suggests a protective role of this natural barrier against rabies virus infection in mice. PMID- 10670699 TI - Identification and localization of a schistosome-associated fucosyllactose determinant expressed by Fasciola hepatica. AB - A Biomphalaria alexandrina-derived lectin (BaSII), of proven specificity to a Schistosoma mansoni-associated fucosyllactose [(Fuc alpha1-2) Gal beta1-4 Glc] determinant, was employed to investigate the putative antigenic cross-reactivity between Schistosoma mansoni and Fasciola hepatica in terms of this structurally defined oligosaccharide sequence. BaSII affinity column chromatography of extracts of adult worms metabolically radiolabelled with 35S-methionine and analysis by two-dimensional gels established the expression of the fucosyllactose determinant in multiple copies among heterogeneous, acidic glycoproteins synthesized by adult Fasciola hepatica. Direct fluorescence microscopy revealed that determinant-bearing glycoproteins were localized to the external glycocalyx and perikarya of the tegument as well as the epithelial lining of the intestinal caeca and vitelline ducts and glands. Determinant expression was also evident in embryonated cells of eggs and miracidia as well as the intermediate cellular wall of encysted metacercariae, suggesting its conservation during the course of development of the parasite. Based on the structural relatedness of the cross reactive fucosyllactose determinant to the antigenic mammalian blood group H trisaccharide, our observations may have implications in serodiagnosis and immunoprophylaxis of schistosomiasis/fascioliasis. PMID- 10670700 TI - Identification of a collectin-like protein in pig serum that binds a component in perienteric fluid from Ascaris suum. AB - A collectin-like protein (CLP) of the acute phase protein family that binds the polysaccharides mannan and alpha-1-6 dextran was isolated from the serum of pigs infected with Ascaris suum. A monoclonal antibody generated against this protein and used to characterize the CLP revealed on SDS-PAGE and western blot analysis that the protein had a molecular weight of approximately 48 kDa under reducing conditions and greater than 100 kDa under nonreducing conditions. Enzyme-linked immunosorbent assay (ELISA) showed that the CLP bound to substances in the perienteric fluid of Ascaris suum (APF). Molecular weight fractionation of APF demonstrated that CLP binds primarily to APF substances of greater than 100 kDa. Binding of CLP to APF was partially blocked by phosphatidylinositol. This is the first report of a porcine CLP and the binding of a CLP to components of the common nematode Ascaris suum. PMID- 10670701 TI - Human contact with cattle, dogs and monkeys and genetic diversity of measles viruses. AB - There is a very close contact among humans, cattle, dogs and monkeys in developing countries. That could lead to genetic alteration in morbillivirus isolates of measles, rinderpest, canine distemper. Emergence of novel morbillivirus strains and failures of prophylactic viral vaccines should be monitored by characterisation of genetic profiles of isolates in different species. Incorporation of additional epitopes in existing prophylactic vaccines should minimise vaccine failures attributable to such novel strains. PMID- 10670702 TI - Selenium and vitamin E effect on antibody production of sheep vaccinated against enzootic abortion (Chlamydia psittaci). AB - The effect of selenium (Se) and vitamin E (vit E) on antibody production of sheep vaccinated against Chlamydia psittaci (ovis) was investigated. Thirty-two sheep, one year old, seronegative to Chlamydia infection, vaccinated against enterotoxemia and dewormed were used. Injectable sodium selenite (0.1 mg/kg b.w.) was given twice to animals of the first group (gSe), with a three week interval. The sheep of the second group (gE) received 1 g vit E each orally, six times at weekly intervals. The animals of the third group (gSeE) were given Se and vit E in doses and routes of administration as in gSe and gE. The animals of the fourth group served as controls (gC) and injected normal saline. The first vaccination was made at the time that the second Se injection was given. Revaccination was made two weeks later. The experiment lasted 29 weeks. The results indicated that Se alone led to a significant increase of Chlamydia antibody response (P < 0.05), but not when it was given in combination with vit E. Animals that received vit E (gE) had much lower titres, just above of those of the controls. PMID- 10670703 TI - Anxiolytic 2,3-benzodiazepines, their specific binding to the basal ganglia. AB - Over the past 20 years, several members of the 2,3-benzodiazepine family have been synthesized. Some of these compounds--tofisopam (Grandaxin), girisopam, nerisopam--exert significant anxiolytic and antipsychotic activities. Sites where actions of 2,3-benzodiazepines are mediated differ from those of 1,4 benzodiazepines. Binding of 2,3-benzodiazepines to neuronal cells in the central nervous system shows a unique and specific distribution pattern: their binding sites are located exclusively to the basal ganglia. Chemical lesioning of the striato-pallido-nigral system, surgical transections of the striato nigral pathway and the activation of c-fos expression in the basal ganglia after application of 2,3-benzodiazepines suggest that these compounds mainly bind to projecting neurons of the striatum. The binding sites are transported from the striatum to the substantia nigra and the entopeduncular nucleus. Recent studies on mechanism of action of 2,3-benzodiazepines indicate their possible role in opioid signal transduction since 2,3-benzodiazepines augment the agonist potency of morphine to induce catalepsy and analgesia, and their action is diminished in morphine tolerant animals. The possible biochemical target of 2,3-benzodiazepines is an alteration in the phosphorylation of protein(s) important in the signal transduction process. Agents affecting emotional responses evoked by endogenous opioids without danger of tolerance and dependence may represent a new therapeutic tool in the treatment of addiction and affective disorders. PMID- 10670704 TI - Ethanol and amino acids in the central nervous system: assessment of the pharmacological actions of acamprosate. AB - Ethanol induces alterations in the central nervous system by differentially interfering with a number of neurotransmitter systems, although the mechanisms by which such effects are executed are not well understood. The present review therefore, is designed to ascertain the effect of ethanol on both excitatory and inhibitory amino acid neurotransmitters, as well as the sulphonated amino acid taurine, assayed by the microdialysis technique within specific brain regions of rat during different types of alcohol intoxication, acute and chronic, as well as during the withdrawal period. Such an understanding of these pharmacological actions of ethanol on neurotransmitters is essential in order to provide the impetus for the development of appropriate therapeutic intervention to ameliorate the multitude of neurochemical disorders induced by ethanol. In addition the possible mode of action of a new therapeutic drug for the treatment of alcoholism, acamprosate will be discussed. The first part of this review will be limited to studies of the effect of ethanol on both amino acid neurotransmitters and the sulphonated amino acid taurine, a possible neuromodulator. While, the second part will seek to establish the possible mechanism of action of a new therapeutic drug, acamprosate, which is used to combat the effects of ethanol, particularly during the craving period, as well as maintaining abstinence in weaned alcoholics. PMID- 10670705 TI - Presenilins and Alzheimer's disease: biological functions and pathogenic mechanisms. AB - Alzheimer's disease (AD) is the most common cause of dementia in the elderly population. Dementia is associated with massive accumulation of fibrillary aggregates in various cortical and subcortical regions of the brain. These aggregates appear intracellularly as neurofibrillary tangles, extracellularly as amyloid plaques and perivascular amyloid in cerebral blood vessels. The causative factors in AD etiology implicate both, genetic and environmental factors. The large majority of early-onset familial Alzheimer's disease (FAD) cases are linked to mutations in the genes coding for presenilin 1 (PS1) and presenilin 2 (PS2). The corresponding proteins are 467 (PS1) and 448 (PS2) amino-acids long, respectively. Both are membrane proteins with multiple transmembrane regions. Presenilins show a high degree of conservation between species and a presenilin homologue with definite conservation of the hydrophobic structure has been identified even in the plant Arabidopsis thaliana. More than 50 missense mutations in PS1 and two missense mutations in PS2 were identified which are causative for FAD. PS mutations lead to the same functional consequence as mutations on amyloid precursor protein (APP), altering the processing of APP towards the release of the more amyloidogenic form 1-42 of Abeta (Abeta42). In this regard, the physical interaction between APP and presenilins in the endoplasmic reticulum has been demonstrated and might play a key role in Abeta42 production. It was hypothesized that PS1 might directly cleave APP. However, extracellular amyloidogenesis and Abeta production might not be the sole factor involved in AD pathology and several lines of evidence support a role of apoptosis in the massive neuronal loss observed. Presenilins were shown to modify the apoptotic response in several cellular systems including primary neuronal cultures. Some evidence is accumulating which points towards the beta-catenin signaling pathways to be causally involved in presenilin mediated cell death. Increased degradation of beta-catenin has been shown in brain of AD patients with PS1 mutations and reduced beta-catenin signaling increased neuronal vulnerability to apoptosis in cell culture models. The study of presenilin physiological functions and the pathological mechanisms underlying their role in pathogenesis clearly advanced our understanding of cellular mechanisms underlying the neuronal cell death and will contribute to the identification of novel drug targets for the treatment of AD. PMID- 10670706 TI - Endothelin and dopamine release. AB - Endothelins and endothelin receptors are widespread in the brain. There is increasing evidence that endothelins play a role in brain mechanisms associated with behaviour and neuroendocrine regulation as well as cardiovascular control. We review the evidence for an interaction of endothelin with brain dopaminergic mechanisms. Our work has shown that particularly endothelin-1 and ET(B) receptors are present at significant levels in typical brain dopaminergic regions such as the striatum. Moreover, lesion studies showed that ET(B) receptors are present on dopaminergic neuronal terminals in striatum and studies with local administration of endothelins into the ventral striatum showed that activation of these receptors causes dopamine release, as measured both with in vivo voltammetry and behavioural methods. While several previous studies have focussed on the possible role of very high levels of endothelins in ischemic and pathological mechanisms in the brain, possibly mediated by ET(A) receptors, we propose that physiological levels of these peptides play an important role in normal brain function, at least partly by interacting with dopamine release through ET(B) receptors. PMID- 10670707 TI - Modeling of separations by closed-loop steady-state recycling chromatography of a racemic pharmaceutical intermediate. AB - Closed-loop steady-state recycling (SSR) is a cyclic, one-column process that is similar to simulated moving bed (SMB) chromatography in several respects. Both processes are cyclic. In both SMB and SSR, fractions are collected from the leading and trailing portions of the circulating chromatographic profile, and fresh sample is injected into the interior of the profile. However, SMB is a continuous process whereas SSR is a discontinuous, repetitive process. This paper presents a model for the closed-loop SSR process and its experimental validation in a case of practical importance. For this last purpose, we used the closed-loop SSR separation of the enantiomers of a racemic pharmaceutical intermediate. The experimental determination of the competitive adsorption equilibria was performed by frontal analysis in a system composed of a chiral HPLC column as the stationary phase and pure acetonitrile as the mobile phase. All the adsorption data were well correlated by the Langmuir model. The Langmuir model was used to calculate overloaded band profiles corresponding to the separation of racemic mixtures at both analytical and preparative scales. Theoretical band profiles were calculated using the equilibrium-dispersive model. With proper corrections for the contributions of the sources of extra-column band broadening, the model properly predicts the experimental band profiles obtained in the closed-loop SSR setup and demonstrates that a cyclic steady-state develops after the completion of a finite number of cycles. The results also show that the extra-column effects must be accounted for in order to model the closed-loop SSR process accurately. PMID- 10670708 TI - Simultaneous optimization of the analysis time and the concentration detectability in open-tubular liquid chromatography. AB - Scott's OT-LC minimal analysis time problem has been solved analytically and has been extended to thick-film and/or large diameter columns. The optimisation analysis has also been applied to a number of relative performance indexes (Cmax/t(anal), Cmax x d/t(anal) and Cmax x u x d2/t(anal) which provide a quantitative insight on the extent to which OT-LC allows to combine short analysis times with a large concentration detectability. PMID- 10670709 TI - Dissociation constants of phenols in methanol--water mixtures. AB - A preferential solvation model that relates solute properties with solvent composition in binary mixtures has been applied to the dissociation pKa values of a set of 28 substituted phenols in methanol-water mixtures. The parameters of the model allow estimation of the pKa value of each phenol for any methanol-water composition. Moreover, it is demonstrated that the pKa values of the whole set of phenols at any methanol-water composition are linearly related to the pKa values of the phenols in water. Equations that relate the correlations' slope and intercept values with the solvent composition have been derived and tested with the set of phenols. The general parameters obtained for these equations allow an accurate calculation of the pKa value of any phenol, even of those not included in the original set, at any methanol-water composition solely from the pKa value of the phenol in water. These calculated pKa values can be used for quantitative structure-HPLC retention relationships. The method is tested by comparison of the calculated pKa values with the HPLC determined pKa values of 26 phenols in a polymeric column with a 50% methanol as mobile phase. PMID- 10670710 TI - Ion exchange on resins with temperature-responsive selectivity IV. Influence of solution and column parameters on efficiency of reagentless separation of copper and zinc using thermo-induced concentration waves technique. AB - The effect of solution and column parameters such as pH, ratio of metal ions and the height of resin bed on formation of the thermo-induced concentration waves (TICW) in the course of the thermostripping process of Cu2+ and Zn2+ ions from iminodiacetic resin Lewatit TP-207 has been studied. The formation of TICW is attributed by both different diffusivities of metal ions and the presence of the third (H+) ion in the system. This effect makes it possible to carry out a reagentless separation of metal ions by using either the metal depletion or metal concentration mode of operation. The efficiency of the TICW-based separation process by using the metal depletion mode appears to be the higher the lower relative concentration of metal ion under removal (slower diffusing ionic species). The mathematical model proposed allows for qualitative and quantitative description of thermostripping processes in a wide range of pH and solution compositions. The physical model of the TICW process proposed makes it possible to account for the influence of different process parameters and generalise the applicability of the TICW separation technique to the binary mixtures of any metal ions with different diffusivities. PMID- 10670711 TI - Visualization of solute migration in chromatographic columns quantitation of the concentration in a migrating zone. AB - The concentration distribution across a zone of iodine migrating along a column made of glass, packed with C18-bonded silica, and eluted with carbon tetrachloride was derived from a quantitative analysis of the photographs of the zone. The photographs were scanned and turned into digital images. The intensity distributions obtained from the measurement of the grayscale intensity were converted into concentration profiles using a calibration method. This procedure is illustrated and suitable corrections are introduced to account for the transverse variation of the optical path length, as a result of using a cylindrical detector cell (the column itself), and for the refraction of light due to the differences between the refraction indices of the glass wall and the liquids involved. An error analysis is also reported. It shows that the method can reliably produce results with a precision of a few percent, allowing on column evaluation of column performance and the derivation of the radial distributions of the column efficiency, the migration velocity of the zone, and the sample distribution at the head of the column. PMID- 10670712 TI - High-sensitivity chromatographic detector incorporating three-dimensional charge coupled device fluorimetry. AB - This article describes a detector for liquid chromatography that provides three dimensional, excitation-emission-intensity data of fluorescent compounds. The instrument achieves multi-wavelength excitation by employing a polychromator that disperses radiation along the vertical portion of a quartz capillary flow cell. The ensuing sample fluorescence is collected in the horizontal plane and dispersed using an emission polychromator to form a focused two-dimensional excitation-emission image at a charge coupled device (CCD) pixel array. Each vertical pixel equates to a narrowly defined excitation wavelength band, whilst horizontal pixels equate to corresponding emission wavelengths, with the intensity of light encroaching onto the CCD pixel elements providing the third dimensional variable in the measurement. It is shown that the extreme sensitivity of CCD detection coupled to the dual polychromator arrangement makes possible the collection of fluorescence excitation-emission-intensity data of polycyclic aromatic hydrocarbons in a flow stream in less than 0.05 s with nanogram per millilitre limits of detection. PMID- 10670713 TI - Size-exclusion chromatography--multiangle laser light scattering analysis of beta lactoglobulin and bovine serum albumin in aqueous solution with added salt. AB - The solution characteristics of beta-LGB (beta-lactoglobulin) and BSA (bovine serum albumin) are reported as determined by size-exclusion chromatography with on-line multiangle laser light scattering, differential refractive index and UV detection. The order of the three in series placed detectors as well as the interdetector volumes have been carefully pointed out. At concentrations below 2.5 mg/ml and at different values of pH the weight-average molecular mass of both proteins have been obtained. They indicate the appearance of monomers, dimers and higher order multimers. For beta-LGB the growth of self-associates could be observed at the isoelectric point over a period of days. The range of applicability of the method is discussed. PMID- 10670714 TI - Enhanced performance of expanded bed chromatography on rigid superporous adsorbent matrix. AB - Rigid spherical macroporous adsorbent beads with surface hydroxyl groups were prepared by cross-linking of cellulose. These beads had diameter in the range 100 200 microm and a mean pore size of about 3 microm with about 60% pore volume. The matrix (bulk density approximately 1600 kg m(-3)) could be expanded into a stable bed and used for protein chromatography. Chromatographic runs were performed on a 10 mm diameter column under non-retaining and retaining conditions on the prepared matrix (called Celbeads) and performance of the runs was measured in terms of the height equivalent to a theoretical plate (HETP). The HETP curves in both packed and expanded bed modes followed profiles typical of macroporous adsorbents, i.e. increasing and levelling with velocity. Unimpaired performance of the matrix at increasing flow-rates permitted expanded bed elution of adsorbed solutes without loss of efficiency in terms of purification factor and product concentration. As a model system, Celbeads was used to purify lactate dehydrogenase from porcine muscle homogenate by dye-affinity chromatography. The prepared matrix provided about 100 theoretical plates per meter for the enzyme system at a linear flow velocity of 1.27 cm x min(-1) in an expanded bed elution mode, and gave enzyme yields of 100% with a purification factor of 31 using an optimized procedure. The adsorbent could be cleaned in place with 5 M urea and used repeatedly without loss of performance. PMID- 10670715 TI - Effect of stationary phase hydrophobicity and mobile phase composition on the separation of carboxylic acids in ion chromatography. AB - In a previous work, we studied the retention behavior of monovalent and divalent carboxylic acids on a highly cross-linked polystryene-divinylbenzene anion exchange column (IonPac AS4A-SC) using a carbonate-based buffer, and a retention model was applied to the chromatographic data obtained. In this work we characterized the retention of carboxylates (formic, acetic, propionic, lactic, pyruvic, oxalic, malonic, succinic, fumaric, maleic, tartaric, glutaric, adipic, malic, mucic, trans-beta-hydromuconic, trans,trans-muconic acids) on a column with higher hydrophilicity (IonPac AS11) according to analyte and stationary phase properties, using previously investigated eluent compositions and comparing the retention data obtained. Moreover, the effect of organic modifiers (CH3OH and CH3CN) in the eluent on the retention factors was also evaluated. The chromatographic data obtained on the IonPac AS11 column were fitted by the retention model and allowed one to obtain and to compare ion-specific selectivity constants (parameters of the model) with the ones obtained with the previous column. PMID- 10670716 TI - Comparative methodology in the determination of alpha-oxocarboxylates in aqueous solution ion chromatography versus gas chromatography after oximation, extraction and esterification. AB - The alpha-oxocarboxylates (alpha-ketocarboxylates) and the corresponding alpha oxoacids (alpha-ketoacids) have been reported as byproducts of ozonation of potable water supplies. Some of these species also occur in biophysiological systems. Five analytes were investigated in this study: oxoethanoate (glyoxylate), 2-oxopropanoate (pyruvate), 2-oxobutanoate (2-ketobutyrate), 2 oxopentanoate (2-ketovalerate) and oxopropanedioate (ketomalonate, mesoxalate). Ion chromatography (IC) and gas chromatography (GC) were evaluated for the quantitation of these analytes at concentrations < or =200 ng ml(-1). For the IC method, the samples are run directly with minimal to no pre-treatment. For the GC method, the analytes must be derivatized with O-(2,3,4,5,6 pentafluorobenzyl)oxylamine to form oximes. The oximes are extracted into tert butyl methyl ether and the carboxylic acid is esterified (methylated) with diazomethane. It was concluded that the ion chromatographic determination is significantly superior to the gas chromatographic method for these analytes. PMID- 10670717 TI - Peptide map procedure using immobilized protease cartridges in tandem for disulfide linkage identification of neu differentiation factor epidermal growth factor domain. AB - Immobilized proteolytic enzyme cartridges were used to rapidly digest neu differentiation factor EGF domain in order to obtain improved peptide maps useful for assignment of disulfide linkages. The procedure described here involves an on line digestion of proteins using immobilized trypsin and endoproteinase Glu-C cartridges connected in series, followed by on-line RP-HPLC separation of the peptides. The entire process can be automated using a commercially available workstation; and the total time required for both proteolytic digestion and the HPLC separation can be shortened to within 1 h. Using these immobilized columns, we demonstrated that disulfide structure assignment of the EGF domains of recombinant human neu differentiation factor can be performed by isolation of individual disulfide-containing peptides followed by assignment of disulfide linkages with prompt fragmentation of peptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The use of immobilized protease cartridges in tandem eliminates undesirable digestion artifacts associated with longer digestion time and higher protease-to-substrate ratio and results in the development of a reproducible and high quality peptide map. PMID- 10670718 TI - New affinity nylon membrane used for adsorption of gamma-globulin. AB - Microporous polyamide membranes were first modified by acid hydrolysis and subsequently bound with hydroxy-ethylcellulose to amplify reactive groups and reduce nonspecific interactions with proteins. Then 1,6-diaminohexane as space arm and phenylalanine as ligand were immobilized onto the nylon membranes by s triazine trichloride activation. Affinity membranes thus obtained were set in a stack and used to adsorb gamma-globulin. The adsorption capacity (qm) of the affinity membrane is 53 micro gamma-globulin per m2 membrane and the desorption constant (Kd) is 2.35 x 10(-6) mol/l. The effects of feed, washing and elution rates on adsorption and desorption behavior were investigated. The results showed that affinity purification through these membranes could not be operated at very high flow-rates. A stack of 20 membranes with 47 mm diameter can adsorb 7.8 mg gamma-globulin with a purity of 91.6% from 4 ml of human plasma in a single-pass mode. PMID- 10670719 TI - Determination of phenoxyalkanoic acids and other herbicides at the ng/ml level in water by solid-phase extraction with poly(divinylbenzene-co-N-vinylpyrrolidone) sorbent and high-performance liquid chromatography--diode-array detection. AB - A method for the determination of phenoxyalkanoic acids and other polar compounds in environmental water samples without pH adjustment before extraction has been developed. Recoveries were calculated from 500 ml of milliQ water spiked at the level of 0.5 ng/ml using solid-phase extraction (SPE) and HPLC-DAD. Different SPE materials (RP-C18, ENV+, ENV+-C8, SAX and Oasis HLB) were tested. After method optimization, 15 of the 16 compounds studied could be extracted with recoveries better than 70% on the most suitable copolymeric poly(divinylbenzene-co-N vinylpyrrolidone) material (Oasis HLB cartridges). PMID- 10670720 TI - Synthesis and characteristics of [60]fullerene polysiloxane stationary phase for capillary gas chromatography. AB - A new type of fullerene-containing polysiloxane was synthesized by reacting [60]fullerene with azidopropyl polysiloxane directly. The polysiloxanes have been used successfully as stationary phases for capillary gas chromatography. They displayed high column efficiency, wide operational temperature and high thermostability, and exhibited unique selectivity for many organic substances, such as alkanes, alcohols, ketones and aromatic compounds. The stationary phase was especially suitable for separation of high boiling-point compounds like polycyclic aromatic hydrocarbons and phthalic esters, etc. It was also found that some alcoholic or aromatic positional isomers could be well separated on the column. The influence of the fullerene content on the separation was also investigated. PMID- 10670721 TI - Identification of compounds and specific functional groups in the wavelength region 168-330 nm using gas chromatography with UV detection. AB - A GC-UV instrumental set up with two different GC units has been used for determination of specific functional groups and compounds in complex mixtures. Separations have been made using a micro gas chromatograph built into a gas flow cell and by means of an external capillary gas chromatograph linked to the same gas flow cell. Four various applications (cigarette smoke, petroleum, dust, flavour) have been performed in order to demonstrate the potential of the GC-UV method. Gas phase UV spectra have been recorded in the region of 168-330 nm. Based on a gas phase spectrum reference library the identification of unknowns as well as the determination of specific functional groups have been achieved. A table showing the spectral shapes and positions of the absorption bands for 50 specific functional groups is presented. The advantage of using derivative spectra in order to amplify spectral details and improve selectivity is discussed. Regarding sensitivity, it has been found that identifications can be made in the mid-pg range and limit of detection for naphthalenes are at a level of 0.5-3 pg/s. PMID- 10670722 TI - On-line coupling of equilibrium-sorptive enrichment to gas chromatography to determine low-molecular-mass pollutants in environmental water samples. AB - On-line combination of equilibrium sorptive enrichment and gas chromatography is used for the analysis of a group of pollutants varying widely in polarity and volatility in aqueous samples at trace levels. For the ESE process open-tubular traps were used. The newly developed hyphenated method shows a high sensitivity for all the compounds under study. The detection limits were typically between 0.1 and 1 microg/l. The sample volumes required for the compounds to reach equilibrium with the stationary phase are in the range of 20 ml for the aromatic hydrocarbons included in the study (benzene, toluene and p-xylene), to 200 ml for epichlorohydrin and dichlorohydrin. Within- and between-day precision of the absolute peak areas varied between 3 and 16%. The performance of the new method was tested by the analysis of different environmental water samples. PMID- 10670723 TI - Ultra trace detection of a wide range of anabolic steroids in meat by gas chromatography coupled to mass spectrometry. AB - The control on use of anabolic agents in meat producing animals is generally based on urine, faeces or hair analysis. This exercise, which is usually performed in slaughterhouses or on farms, is not relevant to imported carcasses or retail meat. A single sensitive method for a wide range of anabolic steroids was developed. After extraction of the lyophilised meat, enzymatic hydrolysis was used for deconjugation. Solid-phase extraction on a polymeric stationary phase was performed prior to hydrolysis of ester residues under alkaline conditions. Liquid-liquid partitioning was used to separate the analytes into two main categories: phenol containing molecules, such as phenolic steroids, resorcylic acid lactones and stilbenes, and delta4-3-one containing molecules, such as most androgens and progestagens. Solid-phase extraction on silica columns was performed before applying a specific derivatisation for each compound sub-group. The combination of high-resolution chromatography with a quadrupole mass spectrometer permitted detection of 23 steroids in the 5-100 ng/kg range. Ion chromatograms for residue positive samples are shown and discussed. PMID- 10670724 TI - Pesticide trace analysis using solid-phase extraction and gas chromatography with electron-capture and tandem mass spectrometric detection in water samples. AB - Gas chromatography (GC) with electron-capture detection (ECD), mass spectrometry (MS) and tandem mass spectrometry (MS-MS) were employed for the identification of 12 pesticides in water samples. For this purpose, a solid-phase extraction procedure with C18 cartridges was used, optimising the breakthrough volume and the saturation concentration. In GC-MS-MS, the lowest detectable concentrations for the pesticides were between 2 and 26 ng l(-1), recoveries ranged from 70 to 133% in water samples spiked at 100 ng l(-1) and the relative standard deviations were in the range 5.3 to 17.4%. The proposed analytical methodology was applied to analyse pesticides in wetland samples from Almeria (Spain). PMID- 10670725 TI - Resolution optimisation in micellar electrokinetic chromatography using empirical models. AB - Theoretical and empirical models can be used to model the migration or separation characteristics in micellar electrokinetic chromatography in order to optimise the resolution. In this paper only empirical models were used, because it is easier and more straightforward to obtain these models. Several empirical approaches for the optimisation of the resolution were compared in order to determine which response should be modelled preferably. The use of models of the effective mobility in combination with average plate numbers proved to be the most suitable approach to optimisation of the resolution, because the relative prediction errors of the models of the effective mobility were a factor of 2-4 smaller than the relative prediction errors of the models of the apparent mobility. Moreover for the least separated peak pair the resolutions based on the models of the apparent and effective mobility showed relative prediction errors that were approximately a factor of 2 smaller than the relative prediction errors of the resolutions based on the models of the resolution and separation factor. The predictions of the separation factor based on the different models generally showed lower prediction errors than the predictions of the corresponding resolutions. Although the relative prediction errors were large, particularly for closely migrating compounds, the empirical approach will probably lead to the optimum separation buffer composition. PMID- 10670726 TI - Simultaneous determination of polycarboxylic acids by capillary electrophoresis with a copper electrode. AB - The simultaneous determination of polycarboxylic acids including oxalic acid, citric acid, malonic acid, malic acid, tartaric acid, aspartic acid and glutamic acid was achieved by capillary electrophoresis with a copper disk electrode (d = 200 microm). In the system. 0.2 mmol/l cetylpridinium bromide (CPB) was used as an electroosmotic flow (EOF) modifier to reverse the direction of EOF. The effects of the solution pH and CPB concentration on separation were evaluated to achieve the optimum separation conditions. At the working potential of +0.14 V (vs. saturated calomel electrode), the calibration curves for all polycarboxylic acids studied were linear with 2 approximately 3-orders of magnitude and all the detection limits (S/N = 3) were below 15 fmol except malonic acid. Furthermore, the oxalic and citric acids in urine were successfully separated and determined with high sensitivity. PMID- 10670727 TI - Microchip capillary electrophoresis using on-line chemiluminescence detection. AB - Chemiluminescence detection was used in capillary electrophoresis integrated on a microchip. Quartz microchips have two main channels and four reservoirs. Dansyl lysine and -glycine were separated and detected with bis[(2-(3,6,9 trioxadecanyloxycarbony)-4-nitrophenyl]oxalate as peroxyoxalate chemiluminescent reagent. These dansyl amino acids came into contact with the chemiluminescence reagent to produce visible light at the interface between the separation channel and chemiluminescence reagent-containing reservoir. The detection limit (S/N = 3) for dansyl-lysine was 1 x 10(-5) M, which corresponded to the very small mass detection limit of ca. 0.4 fmol. However, the concentration sensitivity in the present system was approximately two orders of magnitude lower than that in the conventional capillary electrophoresis-chemiluminescence detection system. The relative standard deviations of migration time and peak height for dansyl-lysine were 4.2 and 4.5%, respectively. A channel conditioning before every run and an appropriate control of voltages were needed for the reproducible results. The present system had advantages in rapid separation time (within 40 s), small (several 10 pI) and accurate sample injection method using a cross-shaped injector, and simplification and miniaturization of the detection device. PMID- 10670728 TI - Determination of phenols using simultaneous steam distillation-extraction. AB - Simultaneous distillation-extraction was proposed as a preconcentration step for the determination of phenol and its derivatives in aqueous and soil samples. Detection limits of 0.01 mg l(-1) (water) and 0.1 mg kg(-1) (soil) were achieved by gas chromatography-flame ionization detection. The described preconcentration procedure was applied for the primary study of the adsorption equilibrium in a water-soil system serving as a model of phenol behaviors in the environment. PMID- 10670729 TI - Development of a protocol for the automated analysis of amino acids in brain tissue samples and microdialysates. AB - An automated precolumn derivatisation method has been developed for the measurement of fourteen amino acids in brain tissue and microdialysate samples. The method involves labelling amino acids with naphthalene-2,3-dicarboxaldehyde (NDA) in the presence of cyanide (CN-). The resulting highly stable N-substituted 1-cyanobenz[f]isoindole (CBI) derivatives were separated using a binary gradient elution profile and detected fluorometrically. The order of elution of the derivatised amino acids was confirmed by using liquid chromatography with fluorescence and mass spectrometric detection in tandem. Linear calibration plots were obtained for all amino acids in the range studied (0.2-12.5 microM). The limit of detection for CBI derivatives of amino acids was in the range 5-20 fmol (S/N=2) using a 5 microl injection volume. The method has been used for the measurement of amino acids in microdialysates from rat brain and tissue homogenates from different regions of mouse brain. PMID- 10670730 TI - Simultaneous determination of the enantiomers of leucine and [2H7]leucine in plasma by capillary gas chromatography-mass spectrometry. AB - A method for the stereoselective assay of D- and L-enantiomers of both leucine and [2H7]leucine in rat plasma was developed using gas chromatography-mass spectrometry-selected-ion monitoring. DL-[2H3]leucine was used as an internal standard. The method involved purification by cation-exchange chromatography using BondElut SCX cartridge and derivatization with hydrochloric acid in methanol to form methyl ester followed by subsequent chiral derivatization with (+)-alpha-methoxy-alpha-trifluoromethylphenylacetyl chloride to form diastereomeric amide. The derivatization made the separation of the leucine enantiomers possible with good gas chromatographic behavior. Quantitation was performed by selected-ion monitoring of the quasi-molecular ions of the diastereomers on the chemical ionization method. The sensitivity, specificity, accuracy and reproducibility of the method were demonstrated to be satisfactory for application to pharmacokinetic studies of leucine enantiomers. PMID- 10670731 TI - Determination of dihydroergotamine in human plasma by high-performance liquid chromatography with fluorescence detection. AB - Dihydroergotamine, a 5-hydroxytryptamine antagonist, is used for the treatment of vascular headaches. A high-performance liquid chromatography assay with fluorescence detection is described for the determination of dihydroergotamine in plasma. The assay was validated over the concentration range 0.1-10 ng/ml plasma and applied to the analysis of plasma samples from subjects treated intramuscularly and intranasally with 2 mg of dihydroergotamine. PMID- 10670732 TI - Determination of nelfinavir, a potent HIV protease inhibitor, and its active metabolite M8 in human plasma by high-performance liquid chromatography with photodiode-array detection. AB - We developed and characterized a high-performance liquid chromatographic assay for the determination of nelfinavir (NFV), a potent HIV protease inhibitor, and its active metabolite M8 in human plasma. Extraction of the internal standard, M8 and NFV from the plasma buffered at pH 9.5 was achieved by a liquid-liquid extraction with a mixture of methyl-tert.-butyl ether and hexane. Following two washes of the reconstituted sample with hexane, separation was achieved on an octadecylsilyl analytical column with a mobile phase containing 0.1% trifluoroacetic acid-acetonitrile-methanol (51:46:5, v/v). Detection was performed using an ultraviolet photodiode-array detector. The signal was monitored at a wavelength of 220 nm. The assay was found to be linear and has been validated over the concentration range of 25 to 3000 microg/l for M8 and 25 to 6000 microg/l for NFV, from 500 microl of plasma. Recoveries were 98.9% (SD 8.9%), and 100.2% (SD 11.7%) for M8 and NFV, respectively. Concentrations that gave a signal-to-noise ratio of three (15 microg/l for both M8 and NFV) were selected to determine the limit of detection. The lower limit of quantification (25 microg/l for both M8 and NFV) was defined as the concentration for which the relative standard deviation and the percent deviation from the nominal concentration were lower than 20%. PMID- 10670733 TI - Determination of carnitine and acylcarnitines in biological samples by capillary electrophoresis-mass spectrometry. AB - Free carnitine and acylcarnitines (carnitine esters) play an important role in the metabolism of fatty acids. Metabolic disorders can be detected by abnormal levels of these compounds in biological fluids. Capillary electrophoresis-mass spectrometry has the advantage of combining an efficient separation technique with highly selective detection. Therefore, we have developed a method for the determination of carnitine and several of its esters implementing electrospray capillary electrophoresis-mass spectrometry in the positive ion selected reaction monitoring mode. A sheath-flow interface with a mixture of 2-propanol or methanol, water and acetic acid as sheath liquid and nitrogen as nebulizing gas was used. The zwitterionic analytes migrated as cations in the applied electric field using ammonium acetate-acetic acid or formic acid electrolytes. Separations were performed in aqueous, mixed organic-aqueous and non-aqueous media. The influence of the electrolyte composition on the separation efficiency was investigated. The electrospray conditions have been optimized regarding ion current stability and sensitivity. Ammonium acetate (10 mmol/l)-0.8% formic acid in water or 6.4% formic acid in acetonitrile-water (1:1) were used as running buffers for the determination of carnitine and acylcarnitines in human biological samples. Methanol extracts of dried blood spots were analyzed as well as urine and plasma following sample preparation via solid-phase or liquid-liquid extraction. Recoveries approaching 100% were achieved depending on the analytes and sample preparation procedures employed. Endogenous carnitine and acetylcarnitine were determined at concentrations between 2.7 and 108 nmol/ml in normal human urine and plasma. Other acylcarnitines were detected at levels of below the limit of detection to 12 nmol/ml. Good precision (0.8 to 14%) and accuracy (85 to 111%) were obtained; the achieved limits of quantitation (0.1 to 1 nmol/ml) are sufficient to characterize carnitine and acylcarnitine levels occurring as markers for metabolic disorders. PMID- 10670734 TI - Gas chromatographic-tandem mass spectrometric determination of anabolic steroids and their esters in hair. Application in doping control and meat quality control. AB - We have developed a powerful and simple sensitive method for testing hair for anabolic steroids and their esters. A 100-mg amount of powdered hair was treated with methanol in an ultrasonic bath for extraction of esters, then alkaline digested with 1 M NaOH for an optimum recovery of other drugs. The two liquid preparations were subsequently extracted with ethyl acetate, pooled, then finally highly purified using a twin solid-phase extraction on amino and silica cartridges. The residue was derivatized with N-methyl-N(trimethylsilyl) trifluoracetamide (MSTFA) prior to injection. Analysis was conducted by gas chromatography coupled to a triple quadrupole mass spectrometer. The generally chosen parent ion was the molecular ion while two daughter ions were selected for each compound with collision energies ranging from -16 to -21 eV. Internal standards were nandrolone d3 for non-esterified drugs and testosterone phenyl propionate for esters. The limits of detection calculated from an analysis of the blanks (n=30) were 0.08 pg/mg for nandrolone, 6.20 pg/mg for boldenone, 0.07 pg/mg for methyl testosterone, 0.15 pg/mg for ethinyl estradiol, 2.10 pg/mg for metandienone, 0.86 pg/mg for testosterone propionate, 0.95 pg/mg for testosterone cypionate, 1.90 pg/mg for nandrolone decanoate, 3.10 pg/mg for testosterone decanoate and 4.80 pg/mg for testosterone undecanoate. Application to doping control has been demonstrated. In a series of 18 sportsmen, two tested positive for anabolic steroids in hair whereas urinalysis was negative for both of them. The first positive case was nandrolone and the second case concerned the identification of testosterone undecanoate. Measured in 10 white males aged between 22 and 31 years, the testosterone concentration was in the range 1.7-9.2 pg/mg (mean=5.0 pg/mg). The method was also applied in meat quality control. Of the 187 analyses realized based upon hair and urine sampling in slaughter houses, 23 were positive for anabolic steroids in hair: one case for boldenone, one case for metandienone, two cases for testosterone propionate, three cases for nandrolone, five cases for testosterone decanoate and 11 cases for methyl testosterone. In the meantime, urinalysis was always negative for these drugs or their metabolites. PMID- 10670735 TI - Immunopurification of the blood group RhD protein from human erythrocyte membranes. AB - Rh proteins are membrane proteins encoded by genes at the blood group RH locus. They are of paramount importance in transfusion medicine, but their function is still unknown. Biochemical and biophysical studies of these proteins are scarce since only minute amounts of the very hydrophobic Rh proteins, can be purified from human erythrocytes. Recently, a human monoclonal antibody (LOR-15C9) was described as having the unique property to recognize the Rh30 protein carrying the major blood group D specificity (RhD protein), either in a membrane detergent extract or when blotted on a membrane. In this report, we describe one-step purification of the RhD protein from detergent extracts of red cell membranes, based on immunoaffinity chromatography carried out with immobilized LOR-15C9 IgG. The technique yielded RhD protein with high purity which was devoid of other associated proteins (RhAG, CD47, LW and GPB) that comprise the Rh complex in the erythrocyte membrane. By contrast immunoprecipitation performed with the same antibody led to co-isolation of both RhD and RhAG. PMID- 10670736 TI - Determination of reduced and oxidized homocysteine and related thiols in plasma by thiol-specific pre-column derivatization and capillary electrophoresis with laser-induced fluorescence detection. AB - A new sensitive and rapid capillary electrophoresis (CE) assay for measuring reduced and oxidized thiols in human plasma has been developed. To prevent oxidation of the thiols, whole blood was immediately centrifuged after collection and the plasma proteins were precipitated with perchloric acid. The reduced thiols in the supernatant were derivatized quantitatively at 25 degrees C, pH 7.5 with a fluorescent reagent, fluorescein-5-maleimide (FM). The total plasma concentration of thiols, including the fraction coupled to proteins, was assayed after an initial reduction of the disulfide linkage in plasma with dithiothreitol. The separation of FM-thiols was performed in an acetonitrile/10 mM sodium phosphate-50 mM SDS buffer [25:75 (v/v); pH 7.0] using a fused-silica capillary (57 cm x 75 microm I.D.) at 45 degrees C. A 3-mW argon-ion laser (lambda(ex) 488 nm/lambda(em) 520 nm) was employed for FM-thiol detection. With the electric field of 530 V/cm, the time needed for the separation of FM homocysteine, FM-glutathione and FM-N-acetylcysteine was less than 8 min. The lower limit of detection was 3 microM for the total thiols and 10 nM for the reduced thiols. The method was applied to, the determination of homocysteine levels in plasma from patients with end-stage renal disease. PMID- 10670737 TI - Routine analysis of amphetamine class drugs as their naphthaquinone derivatives in human urine by high-performance liquid chromatography. AB - We describe a simple HPLC method which is suitable for the routine confirmation of immunoassay positive amphetamine urine samples. The precolumn derivisation method employing sodium naphthaquinone-4-sulphonate was found to have adequate sensitivity, selectivity and precision for the measurement of amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4 methylenedioxyamphetamine (MDA), and 3,4-methylenedioxyethylamphetamine (MDEA) at 500 microg/l cutoff level for confirmatory analysis of amphetamines in urine. The specificity of the method is enhanced by detecting the peaks at two different wavelengths. The ratios of the peak heights measured at the two wavelengths were different for each of the 5 amphetamines analysed. There was no interference from other phenylethylamine analogues that are commonly found in "over the counter" preparations. The HPLC method is compared to a commercial TLC system for detecting amphetamines in urine of drug abusers attending drug rehabilitation programmes. The HPLC confirmatory method described is a viable alternative to GC or to the more complex and costly GC-MS techniques for confirming amphetamine, methamphetamine, MDMA, MDA and MDEA in urine of drug abusers especially when used in a clinical care setting. PMID- 10670738 TI - Simple and reliable high-performance liquid chromatography fluorimetric procedure for the determination of amphetamine-derived designer drugs. AB - The paper describes a HPLC-fluorimetric procedure for the determination of methylenedioxyamphetamine, methylenedioxymethamphetamine, methylenedioxyethamphetamine and N-methyl-1-(1,3-benzodioxol-5-yl)-2-butanamine in urine, serum, saliva and street samples, that features interesting advantages over other procedures previously described. The method requires a very small sample volume (100 microl) and no extraction, lacks matrix effect, and is not time consuming. Linearity was in the range 50-1000 ng/ml regardless of matrix. Sensitivity and detection limit were 50 ng/ml and 10 ng/ml, respectively, but they may reach 10 ng/ml and 2 ng/ml if a slight modification is introduced in the procedure. Intra- and inter-day precision were always within 5% and 8%, respectively. Recovery was satisfactory for all matrices. The described procedure could be successfully used for clinical, epidemiological and forensic applications. PMID- 10670739 TI - Simultaneous assay of morphine, morphine-3-glucuronide and morphine-6-glucuronide in human plasma using normal-phase liquid chromatography-tandem mass spectrometry with a silica column and an aqueous organic mobile phase. AB - Morphine (MOR) is an opioid analgesic used for the treatment of moderate to severe pain. MOR is extensively metabolized to morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G). A rapid and sensitive method that was able to reliably detect at least 0.5 ng/ml of MOR and 1.0 ng/ml of M6G was required to define their pharmacokinetic profiles. An LC-MS-MS method was developed in our laboratory to quantify all three analytes with the required sensitivity and a rapid turnaround time. A solid-phase extraction (SPE) was used to isolate MOR, M3G, M6G, and their corresponding deuterated internal standards from heparinized plasma. The extract was injected on a LC tandem mass spectrometer with a turbo ion-spray interface. Baseline chromatographic separation among MOR, M3G, and M6G peaks was achieved on a silica column with an aqueous organic mobile phase consisting of formic acid, water, and acetonitrile. The total chromatographic run time was 3 min per injection, with retention times of 1.5, 1.9 and 2.4 min for MOR, M6G, and M3G, respectively. Chromatographic separation of M3G and M6G from MOR was paramount in establishing the LC-MS-MS method selectivity because of fragmentation of M3G and M6G to MOR at the LC-MS interface. The standard curve range in plasma was 0.5-50 ng/ml for MOR, 1.0-100 ng/ml for M6G, and 10-1000 ng/ml for M3G. The inter-day precision and accuracy of the quality control (QC) samples were <7% relative standard deviation (RSD) and <6% relative error (R.E.) for MOR, <9% RSD and <5% R.E. for M6G, and <3% RSD and <6% R.E. for M3G. Analyte stability during sample processing and storage were established. Method ruggedness was demonstrated by the reproducible performance from multiple analysts using several LC-MS-MS systems to analyze over one thousand samples from clinical trials. PMID- 10670740 TI - Comparison of different elution conditions for the immunopurification of recombinant hepatitis B surface antigen. AB - An immunoaffinity chromatographic method was developed using a mAb immunosorbent to purify recombinant hepatitis B surface antigen (r-HBsAg) from yeast. Elution conditions using a mAb-coated ELISA were improved to select the best conditions to purify r-HBsAg. The optimum results in terms of total quantitative recovery were obtained using 20 mM Tris pH 11.6. An increase in the CB.Hep-1 mAb (anti HBsAg) useful immunosorbents half-life and in its yield per cycle was obtained when alkaline elution conditions were used. Moreover, the basic conditions do not affect either the antigenic characteristics or the purity or the molecular integrity of r-HBsAg. PMID- 10670741 TI - Simple high-performance liquid chromatographic method for the determination of tocotrienols in human plasma. AB - A simple high-performance liquid chromatographic method using fluorescence detection was developed for the determination of vitamin E especially delta-, gamma- and alpha-tocotrienols in human plasma. The method entailed direct injection of plasma sample after deproteinization using a 3:2 mixture of acetonitrile-tetrahydrofuran. The mobile phase comprised 0.5% (v/v) of distilled water in methanol. Analyses were run at a flow-rate of 1.5 ml/min with the detector operating at an excitation wavelength of 296 nm and emission wavelength of 330 nm. This method is specific and sensitive, with a quantification limit of approximately 40, 34 and 16 ng/ml for alpha-, gamma- and delta-tocotrienol, respectively. The mean absolute recovery values were about 98% while the within day and between-day relative standard deviation and percent error values of the assay method were all less than 12.0% for alpha-, gamma- and delta-tocotrienol. The calibration curve was linear over a concentration range of 40-2500, 30-4000 and 16-1000 ng/ml for alpha-, gamma- and delta-tocotrienol, respectively. Application of the method in a bioavailability study for determination of the above compounds was also demonstrated. PMID- 10670742 TI - Measurement of hyperforin a constituent of St. John's wort in plasma by high performance liquid chromatography. AB - Hyperforin is a constituent of Hypericum perforatum extracts (St. John's wort, H. perforatum), which have antidepressant action. Hyperforin was extracted from plasma utilising a solid-phase extraction procedure. Chromatography was performed by isocratic reversed-phase high-performance liquid chromatography with UV end point detection. The calibration curve was linear over the range 0.15-3 microg per ml of plasma. The sensitivity for hyperforin was 4.5 ng on-column. Mean inter and intra-assay relative standard deviations over the range of the standard curve were less than 5%. The absolute recovery for hyperforin averaged 97.8%. PMID- 10670743 TI - Chromatographic approach to study beta-cyclodextrin as a promoter of the penetration of bifonazole into keratinic tissue. AB - A high-performance liquid chromatographic method for the determination of bifonazole in dimethyl sulfoxide solvent was developed to study its penetration into sheephoof. The analytical method was linear over the concentration range studied, i.e., from 0.1 mg/ml to 1 mg/ml. The relative standard deviation was less than 2%. The data obtained showed that complex forming with beta cyclodextrin greatly improved the penetration of bifonazole. PMID- 10670744 TI - Determination of 5-fluorouracil in microvolumes of human plasma by solvent extraction and high-performance liquid chromatography. AB - In the present study, a new reversed-phase HPLC method has been developed and validated for the quantitative determination of 5-fluorouracil (5-FU) in human plasma using only 100-microl samples. The sample extraction and clean-up procedure involved a simple liquid-liquid extraction after addition of 5 chlorouracil (5-CU), used as internal standard, with 5 ml ethyl acetate. Chromatographic separations were performed on an Inertsil ODS-3 column (250x4.6 mm ID; 5 microM particle size), eluted with a mobile phase composed of acidified water (pH 2.0). The column effluent was monitored by UV absorption measurement at a wavelength of 266 nm. The calibration curves were constructed over a range of 0.20-50.0 microM and were fitted by weighted (1/x) linear regression analysis using the ratio of peak heights of 5-FU and 5-CU versus concentrations of the nominal standards. Extraction recoveries over the total range averaged 92 and 93% for 5-FU and 5-CU, respectively. The lower limit of quantitation was established at 0.20 microM (approximately 26 ng/ml), with within-run and between-run precisions of 4.2 and 7.0%, respectively, and an average accuracy of 109.3%. The within-run and between-run precisions at four tested concentrations analyzed in quintuplicate over a time period of four days were < 1.4 and < 4.4%, respectively. The accuracy at the tested concentrations ranged from 98.4 to 102.3%. Compared to previously described validated analytical methods for 5-FU, our present assay provides equivalent to superior sensitivity, using only microvolumes of sample. PMID- 10670745 TI - Quantitative determination of endogenous tetrahydroisoquinoline salsolinol in peripheral blood mononuclear cells by gas chromatography-mass spectrometry. AB - Endogenous 1-methyl-1, 2, 3, 4-tetrahydro-6,7-dihydroxyisoquinoline (salsolinol) could be a potential marker involved in the etiology of alcoholism. The amount of salsolinol analyzed previously from plasma and urine by different methods depends on several dietary conditions because nutrition has an important influence on salsolinol excretion. Whereas plasma salsolinol is influenced by the diet the salsolinol from peripheral mononuclear cells should be endogenously formed. Therefore, a method for the quantification of S-and R-salsolinol from lymphocytes by using gas chromatography-mass spectrometry was developed. The average amount of salsolinol in 10(6) cells was 1.25 ng corresponding to 2.41 x 10(-5) M and was shown to be much higher than the plasma salsolinol concentration (2.6 x 10(-9) M). PMID- 10670746 TI - Type 1 iodothyronine deiodinase in heart --effects of triiodothyronine and angiotensin II on its activity and mRNA in cultured rat myocytes. AB - We previously demonstrated that iodothyronine 5'-deiodination (5'D) activity is present and increased by triiodothyronine (T3) and angiotensin II (Ang II) in cultured rat cardiac myocytes. To further elucidate the stimulatory mechanism of Ang II, we investigated the effect of intracellular Ca2+ and protein kinase C on myocardial 5'D activity. Moreover, to elucidate the molecular mechanism of the stimulatory effect of T3 and Ang II, we detected the mRNA levels by means of a reverse-transcriptase polymerase chain reaction (RT-PCR). 5'D activity was increased by adding Bay-k 8644, Ca2+ channel agonist and the effect of Bay-k 8644 was completely blocked by nifedipine, a Ca2+ channel antagonist. 12-O tetradecanoylphorbol-13-acetate, a protein kinase C activator, similarly stimulated 5'D activity. The addition of a high concentration (20-40 mM) of K+, which caused the depolarization of the membrane had significant stimulatory effects on 5'D activity. Type 1 deiodinase (D1) mRNA was evident in myocardial cells by RT-PCR in a single 758 bp band similar to that in the liver. Cardiac fibroblasts did not express the D1 mRNA. A significant increase in D1 mRNA was also evident after adding T3 and Ang II. These findings indicate that 5'D activity in myocardial cells is increased by activating the voltage sensitive Ca2+ channel, protein kinase C, and membrane depolarization, and that the D1 mRNA is present in cardiac myocytes and is increased by T3 and Ang II. This study therefore suggests that Ang II could affect the action of thyroid hormone on the heart by increasing the D1 gene expression. PMID- 10670747 TI - Insulin resistance contributes to carotid arterial wall thickness in patients with non-insulin-dependent-diabetes mellitus. AB - The aim of this study was to clarify whether insulin resistance contributes to atherosclerosis in patients with non-insulin-dependent diabetes mellitus (NIDDM). Fifty-three NIDDM patients (36 males and 17 females, 53+/-10 years old (mean+/ SD)) were studied. As an index of atherosclerosis, we measured the average thickness (IMT) as well as basal thickness excluding the maximum thickness and the height of the maximum thickness of the carotid artery wall. Euglycemic hyperinsulinemic glucose clamp was conducted for 90 min to evaluate average glucose infusion rate (GIR) as an index of insulin sensitivity in the peripheral tissues. For another 180 min after intake of oral glucose load with 0.3 g/kg, the euglycemic hyperinsulinemic clamp was continued to measure ratio of splanchnic glucose uptake (SGU) as an index of insulin sensitivity of the liver. The patients were separated into three activity groups according to the grade of their leisure-time physical activity. GIR (r = -0.32, p < 0.05) but not SGU (r=0.139) showed a significant inverse relationship with IMT. Multivariant regression analysis indicated that age and total cholesterol remain as independent risk factors for basal thickness and GIR as only independent risk factor for the height of the maximum thickness. Paralleling the degrees of habitual exercise (low, moderate, and high active group), GIR was higher (6.19+/ 1.02, 6.38+/-1.38, 7.44+/-1.80, respectively) and IMT was lower (1.34+/-0.33 mm, 1.20+/-0.31 mm, and 1.12+/-0.29 mm, respectively) in male NIDDM as well as in female NIDDM. These data suggest that insulin resistance in the peripheral tissues but not the splanchnic tissues may independently contribute to carotid arterial wall thickness and especially to plaque lesion, and that habitual exercise might reduce insulin resistance leading to attenuation of atherosclerosis. PMID- 10670748 TI - A Japanese case with Frasier syndrome caused by the splice junction mutation of WT1 gene. AB - The Wilms' tumor suppressor gene, WT1, plays an important role in the development of the urogenital system and also subsequent normal function of this system. Recently, the splice mutations in intron 9 of WT1 gene have been detected in Frasier syndrome, which is characterized by streak gonads, pseudohermaphroditism, slowly progressive nephropathy and frequent development of gonadoblastoma. Here to elucidate the molecular basis in a Japanese patient of Frasier syndrome, WT1 gene was analyzed by polymerase-chain-reaction (PCR) and direct sequencing. We identified the splice junction mutation in intron 9 of WT1, which is recognized as a mutation hot-spot in intron 9. This finding concludes that 1) the mutation in intron 9 might be the cause of Frasier syndrome, and 2) the mutation hot-spot in Japanese and Caucasian patients is similar. PMID- 10670749 TI - The increase of parathyroid hormone-related peptide and cytokine levels in synovial fluid of elderly rheumatoid arthritis and osteoarthritis. AB - We simultaneously measured the concentrations of parathyroid hormone related peptide (PTHrP) and cytokines in synovial fluid (SF) to clarify the relationship between PTHrP and cytokine network in the SF of elderly patients with arthritis. SF was collected from knee joints of five RA patients aged 66+/-11 years old and nine osteoarthritis (OA) patients aged 80+/-9 years old. PTHrP in SF was measured by enzyme-linked immunosorbent assay (ELISA), whereas tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-8 (IL-8) in SF were all measured by ELISA. The PTHrP levels in the SF of RA patients (2.56+/-0.89 pmol/l) were significantly (p<0.05) higher than those of OA patients (1.66+/-0.17 pmol/l). TNF alpha, IL-1beta, IL-2 and IL-6 concentrations in SF of RA were also significantly higher than those in SF of OA (TNF-alpha 22.5+/-14.8 vs 4.8+/-3.0 pg/ml, p<0.01; IL-1beta 11.8+/-11.4 vs 1.4+/-1.3, p<0.05; IL-2 59.9+/-46.6 vs 12.5+/-8.0 pg/ml, p<0.05; IL-6 18424+/-8901 vs 3547+/-2948 pg/ml, p<0.01). The concentrations of IL 4 and IL-8 in SF of RA were similar to those of OA. Immunohistochemical studies revealed the presence of immunoreactive PTHrP in synovial fibroblasts from RA and OA. Among cytokines, only IL-6 was positively correlated with PTHrP levels in SF (r=0.685, p<0.01). In the culture of synovial cells from RA and OA, PTHrP was produced in RA more than OA after phorbol 12-mysistate 13-acetate (TPA) stimulation. These results indicate that PTHrP and cytokines, especially IL-6, might be involved in the inflammatory processes of elderly RA and OA. This is the first study in which PTHrP and cytokine levels were simultaneously examined in synovial fluid of elderly RA and OA. PMID- 10670750 TI - A novel mutation of the KAL1 gene in Kallmann syndrome. AB - Kallmann syndrome is defined by the association of hypogonadotropic hypogonadism and anosmia, for which three modes of transmission have been described: X-linked, autosomal recessive and autosomal dominant. The KAL1 gene, responsible for the X linked form of the disease, has been isolated and its intron-exon organization determined. We report sequence analysis using PCR-direct sequencing method of the entire coding region and splice site junctions of the KAL1 gene in three males with Kallmann syndrome. We found a novel mutation in one case and no mutation in the other two cases. The mutation consisted of a C to T substitution in exon 1 converting codon 66 (CAG) encoding glutamine into a termination codon (TAG)/(Q66X). As a consequence of this mutation, the function of the KAL1 protein consisting of 680 amino acids was severely truncated so as to be consistent with Kallmann syndrome. As only this patient had unilateral renal hypoplasia among the three cases, this would suggest the existence of KAL1 gene mutation in this abnormality. PMID- 10670751 TI - Effects of intravenous administration of high dose-diethylstilbestrol diphosphate on serum hormonal levels in patients with hormone-refractory prostate cancer. AB - The objective of this study was to elucidate the mechanism underlying the further suppression of serum testosterone (T) by diethylstilbestrol diphosphate (DES-DP) in patients with prostate cancer refractory to hormonal treatment. These patients received an LHRH agonist with or without a non-steroidal androgen-receptor blocker or a gestagen before DES-DP. We measured serum levels of total and free T, dihydrotestosterone (DHT), estradiol (E2), dehydroepiandrosterone sulfate (DHEA-S), dehydroepiandrosterone (DHEA), androstenedione, cortisol, aldosterone before and during intravenous administration of high doses of DES-DP (500 or 1000 mg/day). DES-DP administration suppressed the serum levels of FSH (p=0.04) and total T (p=0.02), and eliminated free T (p=0.04) and E2 (p=0.04) from serum, while reducing serum DHEA-S to approximately two-thirds of the pretreatment level (p=0.03). In contrast, serum levels of SHBG (p=0.02) and cortisol (p=0.02) were markedly increased after DES-DP administration. The latter had no significant effect on serum levels of LH, DHT, ACTH, 17alpha-hydroxypregnenolone, 17alpha hydroxyprogesterone, DHEA, androstenedione, or aldosterone. The results suggest that the potent suppression of circulating total T by DES-DP is caused, in part, by the inhibitory effect of DES-DP on serum DHEA-S level. In most patients, high dose DES-DP treatment completely suppressed the serum level of free T, while possibly elevating serum SHBG and decreasing serum total T. The mechanisms that maintain the serum level of serum DHT during DES-DP treatment require further elucidation. PMID- 10670752 TI - Serum leptin levels and bioelectrical impedance assessment of body composition in patients with Graves' disease and hypothyroidism. AB - We investigated whether thyroid status modulates serum leptin concentrations and body composition as determined by bioelectric impedance analysis (BIA). The percent body fat mass (%FM) in male Graves' disease was significantly lower than that in age- and sex- matched normal subjects, at the levels of 11.4+/-6.4% (mean+/-SD) vs 19.9+/-9.2% for men (n=12, P<0.05) but not for women (22.6+/-7.6% vs 24.9+/-13.1%, n=28). In contrast, in female hypothyroidism (n=11) %FM was significantly higher than that in normal subjects (32.9+/-11.5%, P<0.01). Among other body composition parameters, the percentage of body water (%BW), and lean body mass (LBM) were significantly lower in hypothyroid patients, and the ECM (extracellular mass)/BCM (body cell mass) ratio was significantly (P<0.0001) increased in Graves' disease which was the result of marked depletion of BCM with concomitant expansion of ECM. The serum leptin levels were significantly decreased in male Graves' patients (2.3+/-0.7 ng/ml, P<0.05), whereas in female Graves' patients (8.8+/-5.9 ng/ml) and patients with hypothyroidism (9.5+/-7.6 ng/ml), the levels were not different from those of normal controls matched for BMI or %FM. There was a positive correlation between serum leptin levels and %FM in female Graves' patients (r=0.635, P=0.001) and in hypothyroid patients (r=0.801, P=0.014) but not in male Graves patients. There was no significant relationship between serum leptin levels and thyroid hormones, TRAb, or TSAb. In euthyroid obese subjects there was a positive relationship between serum leptin levels and serum TSH levels (r=0.37, P<0.01). These results suggest that hyperthyroidism is characterized by the decreased fat mass and serum leptin levels in men, but female patients appear to be resistant to the effect of thyroid hormones. Together with previous reports, thyroid status has a minor role in the regulation of serum leptin levels. PMID- 10670753 TI - Human thyrotropin-releasing hormone-associated peptide 3 (hTAP-3) in serum. AB - Human thyrotropin-releasing hormone (TRH)-associated peptide 3 (hTAP-3), one of the cryptic peptides resulting from the proteolytic processing of preproTRH to produce TRH, was measured in human plasma from normal, hyperthyroid, and hypothyroid subjects. The dilution curve of hTAP-3 immunoreactivity in the serum paralleled the standard curve of the radioimmunoassay. HPLC analysis revealed a single strong immunoreactive peak, which corresponded to the authentic peptide, hTAP-3. The half-life of hTAP-3 in serum was approximately 3.5 min, and the addition of aprotinin and EDTA completely prevented its degradation. In hyperthyroid patients, plasma concentrations of hTAP-3 were significantly higher than those in the control group and hypothyroid patients, but no correlation was found between its level and that of thyroid hormone. These findings indicate the existence of intact hTAP-3 in the human serum and increases in plasma hTAP-3 levels in hyperthyroid patients, suggesting that blood hTAP-3 may be derived from the peripheral organs rather than the hypothalamus. PMID- 10670754 TI - Hyperphosphatemia accelerates parathyroid cell proliferation and parathyroid hormone secretion in severe secondary parathyroid hyperplasia. AB - We studied the role of phosphorus retention in parathyroid cell proliferation and parathyroid hormone (PTH) oversecretion in severe secondary parathyroid hyperplasia. Mice transplanted with human parathyroid tissue from a patient who had undergone parathyroidectomy for severe secondary hyperparathyroidism were divided into four groups; each group was given a diet with a different phosphorus content (0.4, 0.7, 1.0, and 1.2%) to alter serum phosphorus concentrations. Histologic examinations of grafts by hematoxylin-eosin or by bromodeoxyuridine (BrdU) immunohistochemical staining were performed to assess parathyroid cell proliferation. Changes in serum phosphorus concentrations unidirectionally affected PTH secretion from the graft, because human PTH did not cross-react with mouse PTH. Serum phosphorus concentrations of 1.0P and 1.2P groups were significantly higher than those of 0.4P and 0.7P groups (p<0.05). Serum phosphorus concentrations were significantly correlated with the gradient of human PTH elevation with a coefficient of 0.48 and a p<0.05. Furthermore, serum phosphorus concentrations and the gradient of human PTH elevation were significantly higher in mice with BrdU-immunoreactive cells in the parathyroid graft than in mice without immunoreactive cells in the graft. These results indicate that uncontrolled hyperphosphatemia may accelerate the proliferation of parathyroid cells, exacerbating PTH oversecretion. PMID- 10670755 TI - A case of Hashimoto's thyroiditis with markedly elevated serum thyroglobulin and evidence of its influence on the measurement of anti-thyroglobulin antibody by highly sensitive assays. AB - We present the case of a 66-year-old woman with Hashimoto's thyroiditis, who showed extremely high concentrations of serum thyroglobulin (Tg). Serum Tg levels were markedly elevated following a slight elevation of serum thyrotropin (TSH) (22,000 ng/mL and 11.1 microU/mL, respectively). Although elevated concentrations of serum Tg declined concomitant with decrease of serum TSH one month later, Tg concentrations remained high (> 948 ng/mL) even at normal or suppressed TSH levels. There was no evidence of massive thyroid tissue damage or thyroid tumor. To our knowledge, there have been no case reports of such high concentrations of serum Tg (> 2 x 10(4) ng/mL) in the clinical course of Hashimoto's thyroiditis. Furthermore, we showed evidence that extremely high Tg levels could possibly influence the measurement of anti-Tg autoantibody using highly sensitive radioimmunoassays. PMID- 10670756 TI - cDNA cloning and chromosomal mapping of rat Smad2 and Smad4 and their expression in cultured rat articular chondrocytes. AB - Smad proteins are known to transduce signalling of TGF-beta receptor superfamily. We report here the entire sequences of rat Smad2 and Smad4 which have not been identified yet. Entire sequences were identified by degenerated polymerase chain reaction and following phage library screening and 5' RACE. The predicted amino acid sequences of rat Smad2 and Smad4 are highly conserved among rat, human and mouse. We also mapped these Smads to chromosome 18q.12.3. Unlike endothelial cells, TGF-beta1 stimulates articular chondrocyte proliferation as well as extracellular matrix production, and acts as a repairing agent against cartilage destruction. Since both Smad2 and Smad4 are essential factors for TGF-beta signalling, we examined their expression and regulation in cultured articular chondrocytes. Northern blot analysis showed that TGF-beta1 significantly increased the mRNA level of Smad2 but not of Smad4 in a dose- and time-dependent manner, suggesting that the augmentation of TGF-beta1 action is caused by increasing the expression of the downstream signalling molecule. PMID- 10670757 TI - Changes in serum leptin concentration during behavioral therapy in obese children. AB - To determine the pathophysiological implications of serum leptin level in obesity, we monitored the changes in serum leptin level during outpatient treatment with life style modification in children. Fifty-five obese Japanese children (34 boys and 21 girls; mean age, 9.64 years) were studied. The control children consisted of 42 nonobese subjects (27 boys and 15 girls). The serum leptin concentration was 4.35 +/- 0.46 ng/ml (mean +/- SEM) in the control girls and 2.93 +/- 0.21 ng/ml in the control boys. The serum leptin concentrations in the obese boys and girls were higher than those in their lean counterparts. The concentration in the obese boys (16.28 +/- 1.41 ng/ml) was similar to that in the obese girls (20.33 +/- 2.0 ng/ml). The logarithmic value of serum leptin concentration at the first blood sampling in obese children was correlated with percent overweight and percent body fat. In 36 obese children (24 boys and 12 girls) whose serum leptin concentrations were monitored serially during treatment of obesity, the percent overweight was significantly decreased after the initial sampling. In each individual, the changes in leptin concentration were roughly parallel to those in percent overweight. The ratio of the leptin concentration at the second blood sampling divided by the one at the first sampling in each individual was closely correlated with the respective delta percent overweight. These results suggest that the preceding course of obesity determines the serum leptin level of obese children on longitudinal basis, and that the leptin level reflects the degree of obesity on cross-sectional basis. PMID- 10670758 TI - Effect of pregnancy, lactation and weaning on bone mineral density in rats as determined by dual-energy X-ray absorptiometry. AB - To elucidate the effect of pregnancy, lactation and weaning on bone mineral density (BMD) in rats, a longitudinal study was done on the same individuals measuring BMD by dual-energy X-ray absorptiometry (DXA) and comparing their profiles with those of nonpregnant controls. Twenty-seven pregnant Wistar rats which had been mated at 11 weeks old (baseline), lactated during the three weeks postpartum period and weaned thereafter. Twenty-four rats of the same age served as nonpregnant controls. BMDs in lumbar spine, distal femur and caudal spine of all rats were measured weekly from 11 to 22 weeks except for the week of parturition (14th week). During pregnancy, BMDs of the three sites increased significantly from the baseline values, but no significant difference was observed in comparison with the control. After parturition and during lactation, BMD of the three sites decreased significantly from the pregnant values and decreased even from baseline values. All the BMD values of the pregnant group were significantly lower than those of the control group. After weaning, BMDs of the three sites increased gradually and caught up to the control group at 22 weeks in the lumbar spine and the femur and at 21 weeks in the caudal spine. In conclusion, pregnancy in itself does not significantly affect maternal BMDs of rats, although the significant bone mineral loss during lactation is not completely restored until at least 5 weeks after weaning. PMID- 10670759 TI - Absence of proteolysis of insulin-like growth factor binding protein-3 in serum from patients with growth hormone deficiency. AB - Insulin-like growth factor-I (IGF-I) is predominantly bound to IGF binding protein-3 (IGFBP-3), and free form of IGF-I (fIGF-I) may be bioactive in the circulation. Proteolysis of IGFBP-3, as reported in pregnant serum, results in the lowering of the affinity for IGF-I, thereby increasing the ratio of fIGF-I to total IGF-I (f/t IGF-I ratio). Conflicting results have been reported regarding the relationship between the proteolysis and growth hormone (GH)-IGF-I axis. Proteolysis of IGFBP-3 was previously reported to be present late at night in serum from pediatric subjects with GH receptor dysfunction (GHRD or "Laron-type dwarfism"). Recently, it was reported that proteolysis of IGFBP-3 could not be detected in adult patients with GH deficiency (GHD). The purpose of this study was to investigate the possible relationship between proteolysis of IGFBP-3 and GH in patients with GHD including pediatric cases. Here, proteolysis of IGFBP-3 measured by Western immunoblotting (ages 4-25 years; n=11) and f/t IGF-I ratio measured by immunoradiometric assay (ages 4-25 years; n=10) were studied in patients with GHD, which is similar to GHRD in terms of lowered GH function. There was no significant proteolysis of IGFBP-3 in the sera from the 11 patients with GHD. No proteolysis of IGFBP-3 was observed during a 24 hour period in sera obtained every two hours from two patients with GHD. f/t IGF-I ratio was not increased in plasma from the 10 patients with GHD. Our data suggest that proteolysis of IGFBP-3 is independent of the GH-IGF-I axis. PMID- 10670760 TI - Troglitazone improves insulin-stimulated glucose utilization associated with an increased muscle glycogen content in obese Zucker rats. AB - Recent studies have demonstrated that troglitazone has the capacity to improve insulin resistance. The present study was undertaken to determine the effect of troglitazone on in vivo insulin action, the activities of the pyruvate dehydrogenase (PDH) complex and 3-hydroxyacyl-CoA dehydrogenase (3-HADH) in muscle, and muscle GLUT-4 and glycogen content in obese and lean Zucker rats. Rats were fed a normal chow diet with and without troglitazone as a food admixture (0.2%) for 3 weeks. In vivo insulin action was measured by the sequential euglycemic clamp technique at two different insulin infusion rates (6 and 30 mU/kg BW/min). At the basal (fasting) state and after the clamp studies, the activities of PDH complex and 3-HADH, and the amounts of GLUT-4 and glycogen contained in the red gastrocnemius muscles were determined. Troglitazone treatment produced a significant rise in the metabolic clearance rate of glucose (MCR) during the 6-mU/kg BW/min insulin clamp study (19.5+/-3.9 vs 9.9+/-1.5 ml/kg BW/min, mean+/-SE, P<0.05) in obese rats, but not in lean rats. Troglitazone significantly increased the muscle glycogen content after the clamp study, compared to non-treated rats, in obese rats (9.9+/-0.5 vs 6.5+/-0.4 mg/g tissue, P<0.05) and has the tendency to increase the activity state of PDH complex in obese and lean rats at the fasting state. However, no effect of the drug on muscle GLUT-4 content was found. These results indicate that troglitazone may improve insulin sensitivity associated with increased muscle glycogen content. PMID- 10670761 TI - A case of ectopic thyroid in lateral neck associated with Graves' disease. AB - Thyroid follicles in the lateral position of the neck are usually thought to represent the metastasis of thyroid carcinoma. Here we present a case of a 28 year-old woman with accessory ectopic thyroid associated with Graves' disease. Despite a history of Graves' disease poorly controlled with large dose propylthiouracil she was found to be pregnant and artificial abortion was planned. Thyroid scintigraphy was carried out, which indicated an uptake into the region above the left lobe as well as into both lobes of the thyroid gland. In order to control hyperthyroidism and to exclude the possibility of metastasis, total thyroidectomy with tumor resection was performed before the artificial abortion. Pathological examinations of the thyroid gland indicated findings compatible with Graves' disease. The lateral neck mass was revealed to be composed of nonneoplastic thyroid tissue, showing similar histological findings to those of the goiter, which were consistent with Graves' disease. Taken together with several previous reports, it appears that there are some cases with lateral ectopic thyroid tissue, whose pathogenetic mechanism remains to be elucidated. PMID- 10670762 TI - A novel sex-determining region on Y (SRY) missense mutation identified in a 46,XY female and also in the father. AB - Mutations in the sex-determining gene SR Y previously identified occur in the 46, XY females. In this study, we investigated whether the SR Y mutation participates in the onset of XY sex reversal. Genomic deoxyribonucleic acids (DNA) from five XY sex-reversed females were analyzed for mutations in SR Y using polymerase chain reaction (PCR) amplification and subsequent DNA sequencing. One of the 46, XY females suffered a novel missense mutation at position 306 of SR Y gene, wherein cytosine was replaced by adenine (CGC-->AGC), resulting in a substitution of serine for arginine at amino acid position 76 of SR Y protein. This mutation was located in Helix I of the high-mobility-group (HMG) domain. No other mutations were found in the remaining regions of the gene. Analysis of the SR Y gene in her father revealed that he carried the identical mutation version. This substitution introduces a large basic for a small polar uncharged amino acid residue in the HMG box. The fact that the father transmits the mutant SR Y copy to his offspring implies that SR Y mutations do not always occur in association with sex reversal, even when the ionic environment is altered. PMID- 10670763 TI - An elderly patient with transient diabetes insipidus associated with lymphocytic infundibulo-neurohypophysitis. AB - We present the eldest case ever reported of central diabetes insipidus (DI) associated with infundibulo-neurohypophysitis. A 77-year old woman, who complained of recent development of excessive thirst, polyuria and polydipsia, was referred to our hospital. The daily urine volume was markedly increased to 6 L. DDAVP administration effectively reduced urine volume and increased urine osmolality. The loading test using high-osmolar sodium chloride showed impaired excretion of vasopressin discordant with plasma osmolar changes. The anterior pituitary function was normal. Pituitary magnetic resonance imaging (MRI) showed thickening of the pituitary stalk and a lack of high-intensity signal of the neurohypophysis on T1-weighted images, suggestive of lymphocytic infundibulo neurohypophysitis. The thickness of pituitary stalk on MRI improved 6 months later. DI was controlled with DDAVP for 40 days. This was followed by stabilization of the daily urine volume to less than 2.5 L without DDAVP. Our case is the eldest case of central DI associated with infundibulo neurohypophysitis. The rapid remission of pituitary changes on MRI provides an insight that spontaneously partial remission of central DI may occur, resulting in transient polyuria and polydipsia. PMID- 10670764 TI - Image analysis method to assess adhesion of Helicobacter pylori to gastric epithelium using confocal laser scanning microscopy. AB - We have used confocal scanning microscopy of FITC-labelled bacteria to assess binding of Helicobacter pylori to stomach sections and to assess the effect of inhibitors on binding to the Lewis antigens. We have quantified the binding using an image manipulation package that is readily available on the web. Our results demonstrate heterogeneity of binding of Helicobacter pylori to tissue sections and that binding can be inhibited using synthetic Lewis B oligosaccharide. PMID- 10670765 TI - An enzyme-release assay for the assessment of the lytic activities of complement or antimicrobial peptides on extracellular Toxoplasma gondii. AB - A method is described which allows the evaluation of the membrane lytic activity of either complement or antimicrobial peptides against the extracellular stage of the human protozoan parasite Toxoplasma gondii. The assay is based on lacZ transgenic parasites, determining the activity of released cytoplasmic beta galactosidase into the culture supernatant upon membrane disintegration. This method was used to evaluate the lytic activities of (i) complement which is a natural defense mechanism in infected hosts against extracellular parasites, and (ii) antimicrobial peptides which have not been evaluated against T. gondii before. The results show that the assay provides a simple and convenient way to assess the membrane lytic activity of such compounds and that T. gondii, like other protozoan parasites, is vulnerable to the membrane-lytic effect of antimicrobial peptides. PMID- 10670766 TI - Identification of major subgroups of ammonia-oxidizing bacteria in environmental samples by T-RFLP analysis of amoA PCR products. AB - A cloning-independent method based on T-RFLP (terminal restriction fragment length polymorphism) analysis of amoA PCR products was developed to identify major subgroups of autotrophic ammonia oxidizers of the beta-subclass of the class Proteobacteria in total community DNA. Based on a database of 28 partial gene sequences encoding the active-site polypeptide of ammonia monooxygenase (amoA), defined lengths of terminal restriction fragments (= operational taxonomic units, OTUs) of amoA were predicted to correlate in TaqI-based T-RFLP analysis with phylogenetically defined subgroups of ammonia oxidizers. Members of the genus Nitrosospira showed a specific OTU of 283 bp in length, while a fragment size of 219 bp was indicative of Nitrosomonas-like sequence types including N. europaea, N. eutropha, and N. halophila. Two amoA sequence clusters designated previously as the lineages 'PluBsee' and 'Schohsee' [Rotthauwe, J.-H., Witzel, K.-P., Liesack, W., 1997. Appl. Environ. Microbiol. 63, 4704-4712] shared a TaqI-based OTU with a fragment size of 48 bp, but sequence types of these two lineages could be differentiated by AluI-based T-RFLP analysis. A survey of various environmental samples and enrichment cultures by T-RFLP analysis and by comparative analysis of cloned amoA sequences confirmed the predicted correlations between distinct OTUs and phylogenetic information. Our data suggest that amoA-based T-RFLP analysis is a reliable tool to rapidly assess the complexity of ammonia-oxidizing communities in environmental samples with respect to the presence of major subgroups, i.e. nitrosospiras versus nitrosomonads. PMID- 10670767 TI - Using a green fluorescent protein gene-labeled p-nitrophenol-degrading Moraxella strain to examine the protective effect of alginate encapsulation against protozoan grazing. AB - A gfp-labeled p-nitrophenol-degrading Moraxella strain G21 was used to study grazing of a Tetrahymena thermophila strain in liquid medium. This allowed visualization of the feeding process. Under an epifluorescent microscope, individual G21 fluorescent cells could be seen in vacuoles within the protozoans. Most of the G21 cells appeared to be lysed by T. thermophila and green fluorescent protein released from the bacteria yielded brightly fluorescent food vacuoles inside the protozoans, Examination of population dynamics of a mixed culture of T. thermophila and Moraxella sp. G21 showed that the protozoan reduced the bacterial density from 7.6 to 5.8 log CFU/ml in 2 days. Encapsulating the bacteria in alginate prevented grazing by the protozoans and the density of G21 cells in the beads increased steadily from about 8.3 to 8.9 log CFU/ml in 15 days regardless of the presence of the protozoans. PMID- 10670768 TI - Comparative study of the abundance of various bacterial morphotypes in an eutrophic freshwater environment determined by AODC and TEM. AB - Transmission electron microscopy (TEM) and epifluorescence microscopy were used to obtain comparative measurements of total bacterial counts, and to enumerate abundances of various bacterial morphotypes in an eutrophic freshwater habitat. Although particulate matter would have been expected to interfere with counting by obscuring large areas of the electron microscope grids, estimates of total bacterial abundance made by TEM were on average 1.2 times greater than those obtained using the acridine orange direct counting method (AODC). However, the precision of the AODC method was greater than that for TEM, with a coefficient of variation (C.V.) of 4.0% versus 8.8%, respectively. The total bacterial abundance ranged from 1.1 to 3.2 x 10(6) ml(-1). As was the case for total bacterial density, the numbers of rod- and vibrio-shaped cells were lower when counted in the epifluorescence microscope, indicating the presence of potential starvation forms or ultramicrobacteria. Greatest variations in counts made by TEM and AODC were found for filamentous and coccoid bacteria. Counts of filamentous bacteria made by AODC were only about half of those detected by TEM. In contrast, cocci were on average 1.5 times greater when counted by AODC compared to TEM estimates. Both counting differences were probably caused by the morphology and low density of filamentous and coccoid bacteria (1.7 and 1.4 x 10(5) ml(-1), respectively), which led to an uneven distribution on polycarbonate filters as well as on electron microscope grids. Besides, cocci might easily be mistaken for large viral particles when counted by AODC. Hence, the study supports the use of TEM over AODC for obtaining accurate estimates of total bacterial abundance and especially bacterial morphotypes in natural waters. PMID- 10670769 TI - Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli. AB - A gas chromatographic analysis method was employed to determine the cellular fatty acid (CFA) profiles of spores and vegetative cells of some aerobic endospore-forming bacilli. The harvests of experimental strains were processed to obtain pure spores and acquire whole cell fatty acid methyl esters for the subsequent gas chromatographic analysis, and the corresponding vegetative cells were set as control. Evaluation of reproducibility of spore CFA components revealed that, provided under standardized experimental procedure, spore CFA composition was stable enough for research purposes. Fatty acids recovered in spores in greater quantities were saturated branched-chain acids containing 15 and 17 carbon atoms, similar to the vegetative cells. Commonly, the proportions of saturated branched-chain acids in spores were greater than in vegetative cells. The dendrograms obtained by cluster analysis provided some meaningful taxonomic information of the experimental strains. The fatty acids analysis of spores seems to be a promising supplementary tool for the chemotaxonomic research of aerobic endospore-forming bacilli. PMID- 10670770 TI - A culture apparatus for maintaining H2 at sub-nanomolar concentrations. AB - We devised a microbial culture apparatus capable of maintaining sub-nanomolar H2 concentrations. This apparatus provides a method for study of interspecies hydrogen transfer by externally fulfilling the thermodynamic requirement for low H2 concentrations, thereby obviating the need for use of cocultures to study some forms of metabolism. The culture vessel is constructed of glass and operates by sparging a liquid culture with purified gases, thereby removing H2 as it is produced. We used the culture apparatus to decouple a syntrophic association in an ethanol-consuming, methanogenic enrichment culture, allowing ethanol oxidation to dominate methane production. We also used the culture apparatus to grow pure cultures of the ethanol-oxidizing, proton-reducing Pelobacter acetylenicus (WoAcy 1), and to study the bioenergetics of growth. PMID- 10670771 TI - Assessment of the intracellular pH of immobilized and continuously perfused yeast cells employing fluorescence ratio imaging analysis. AB - The intracellular pH (pHin) of Saccharomyces cerevisiae was measured employing fluorescence ratio imaging microscopy (FRIM). The yeast cells were fluorescently labeled with the pH dependent probe 5(and-6)-carboxyfluorescein (cF) or 5(and-6) carboxyfluorescein succinimidyl ester (cFSE), and subsequently attached to ferric nitrate pretreated glass slides. The labeled and adhered cells could still divide and were metabolically active. Measurement of the pHin was performed during continuous perfusion of the cells with buffer or medium. Cells labeled with cF are highly fluorescent and in non-energized cells the pHin could be easily measured. However, in energized yeast cells cF was accumulated in the vacuoles and/or exported to the extracellular environment, most likely by an energy dependent transport system, thus limiting the time period over which the pHin can be effectively measured. Therefore, cFSE (which conjugates with aliphatic amines in the cytoplasm) was applied to prevent translocation of fluorescent probe to the vacuole and/or extracellular environment. The continuous perfusion in combination with the cFSE labeling of the immobilized cells was successfully applied to determine the effect of low and high pHin and addition of glucose on the pHin of individual yeast cells over a long time period. PMID- 10670772 TI - Use of macroporous polypropylene filter to allow identification of bacteria by PCR in human fecal samples. AB - The detection of pathogenic bacteria directly in human fecal specimens by PCR, requires removal of PCR-inhibitory substances. To investigate whether five different macroporous filters (polypropylene, nylon, polyester, polyethylene, fluorocarbon) could retain polysaccharides, major PCR inhibitors, an in vitro model and human fecal samples were used. The in vitro model consisted of Xanthum gum solutions (3 mg/ml PBS), a bacterial polysaccharide, to which Helicobacter pylori cells were added. Fecal samples from healthy volunteers were spiked with H. pylori and Mycobacterium paratuberculosis cells. Polysaccharide concentrations were significantly reduced only by the polypropylene but not by the other filters. Accordingly, both Xanthum gum solutions and spiked fecal specimens became PCR positive only after filtration with the polypropylene filter. We conclude that this filter can be used to prepare a bacterial DNA template suitable for PCR analysis from human feces. PMID- 10670773 TI - Migraine in childhood--are periodically occurring migraine attacks related to dynamic changes of cortical information processing? AB - Amplitudes and habituation of contingent negative variation (CNV) were analyzed in relation to spontaneously occurring migraine attacks in ten children suffering from migraine without aura. Recording took place during feedback training and instrumental conditioning of slow brain potentials. Both the amplitude of the early CNV component and its habituation deficit increase during the 5 days prior to a migraine attack, with maximum abnormalities the day before the ictal episode. Abrupt reduction of the amplitude and normalization of the CNV habituation were observed during the attack. This study provides evidence for neurophysiological periodicity in young migraineurs and emphasizes that the time relative to the migraine attacks must be considered in studies of juvenile migraine during the headache-free period. PMID- 10670774 TI - Rat oligodendroglia express c-met and focal adhesion kinase, protein tyrosine kinases implicated in regulating epithelial cell motility. AB - Oligodendrocytes, the myelinating cells of the central nervous system, arise from a profilerating pool of motile progenitor cells. The proliferation and survival of these cells is dependent on signal transduction via several protein tyrosine kinases (PTKs) including receptors for fibroblast growth factor -2, the platelet derived growth factor receptors and the neurotrophin receptor, trkC. We hypothesized that additional PTKs could also influence oligodendroglial development. Utilizing RTPCR, we amplified from post-natal day 6 rat oligodendroglia 17 distinct kinase domain sequences, 14 of which were not previously known to be expressed by oligodendroglia. Amongst the sequences identified were the c-met and Fak genes, whose protein products regulate the motility of other epithelial cell types. Utilizing immunohistochemistry, we confirmed that both c-met and Fak are expressed by cultured oligodendroglia, suggesting that these proteins could also be implicated in regulating the motility of these cells. PMID- 10670775 TI - Lithium alters mu-opioid receptor expression in the rat brain. AB - Lithium can potentiate the effects of antidepressant drugs and alters morphine analgesia and phosphoinositide turnover. Analysis of mu-opioid receptor immunostaining after chronic lithium administration in rats revealed an increase in the density of cells expressing mu-opioid receptors in the caudatus-putamen, the dentate gyrus, the lateral septum and the frontal, parietal and piriform cortices. These data suggest that mu-opioid receptor expression in the rat forebrain is altered by in vivo chronic lithium treatment. This could be a compensatory mechanism, induced in part by the effects of lithium on mu-opioid receptor transduction mechanism. PMID- 10670776 TI - A cluster of single nucleotide polymorphisms in the 5'-leader of the human dopamine D3 receptor gene (DRD3) and its relationship to schizophrenia. AB - The association between schizophrenia and the Ser9Gly variant of the dopamine D3 receptor gene (DRD3) has been the subject of numerous studies. Under meta analysis this site, or one or more in linkage disequilibrium with it, appears to contribute a small increase to the relative risk of schizophrenia. In this study, 768 bp of the 5'-leader region of DRD3 mRNA was screened for polymorphisms to assess their contribution to the association of DRD3 with schizophrenia. A cluster of three single nucleotide polymorphisms (SNPs) was identified in tight linkage disequilibrium with each other and with the Ser9Gly polymorphism. One of the 5'-leader SNPs encodes a Lys9Glu variant within a 36 amino acid residue stretch of an upstream open reading frame (uORF). Two common haplotypes are found in the population examined; one is linked to the Ser9 coding variant and the other to the Gly9 variant. A panel of 73 schizophrenic patients and 56 matched controls recruited from the East Anglia region of the United Kingdom was screened for disease association at these sites. Since the 5'-leader and coding sites are in tight disequilibrium, the combined genotype of all 4 sites was scored for each patient. A significant association was seen between disease and the frequency distribution of these genotypes (chi2 = 13.19, d.f. = 3, P = 0.0042; Cochran method for sparse cells applied). A 20% excess of one of the heterozygous genotypes, in which the sequences differ at three of the four SNPs, including Ser9/Gly9 in the receptor and Lys9/Glu9 in the uORF, was found in the patient group. An absence of association of disease with the Ser9Gly polymorphism had previously been reported for this panel. This suggests that these SNPs and the corresponding coding changes may exert a combined or synergistic effect on susceptibility to schizophrenia. PMID- 10670777 TI - Increase of urocortin-like immunoreactivity in the supraoptic nucleus of Dahl rats given a high salt diet. AB - Urocortin-like immunoreactivity (Ucn-LI) in the supraoptic nucleus (SON) of Dahl rats was examined. Dahl salt-sensitive (S) rats fed with a high salt diet developed hypertension. Numbers of Ucn-LI neurons in the SON in Dahl S on a high salt diet were markedly increased, compared with those in Dahl salt-resistant (R) rats on the same. Sporadic Ucn-LI neurons were found in the SON of both Dahl S and R on a normal diet. Numbers of Ucn-LI neurons in the SON of spontaneously hypertensive rat (SHR) and stroke-prone SHR, genetic models of hypertension, and control rats (Sprague-Dawley and Wistar-Kyoto) were similar. These results suggest that Ucn in the SON is associated with salt loading-induced hypertension rather than spontaneous hypertension. PMID- 10670778 TI - Reduced number of striatal neurons expressing preprosomatostatin mRNA in rats with oral dyskinesias after long-term haloperidol administration. AB - Neuroleptic-induced oral dyskinesia in rats, a putative analogue to human tardive dyskinesia, may be due to degeneration within the striatum. Using unbiased stereological methods, a decreased number of striatal neurons expressing preprosomatostatin mRNA was observed only in rats that developed pronounced oral dyskinesias after 30 weeks of haloperidol administration. The amount of preprosomatostatin mRNA in each striatal neuron, measured in terms of optical densities of individual neurons, was not affected by haloperidol. A tendency toward a reduction in the number of NADPH-diaphorase positive neurons was observed in rats receiving haloperidol. These results indicate that the mechanism by which neuroleptics induce oral dyskinesias in rats, and perhaps tardive dyskinesia in humans, involves a functional disruption and possibly damage of a subpopulation of interneurons in the striatum. PMID- 10670779 TI - Early extrastriate activity without primary visual cortex in humans. AB - Damage to the primary visual cortex (V1) destroys the major source of anatomical input to extrastriate cortical areas (V2, V3, V4 and V5) and produces cortical blindness--an absence of any sensation of light and colour--in the visual field contralateral to the side of the lesion. Neuroimaging studies, nevertheless, have recently demonstrated dorsal and ventral extrastriate activation for stationary stimuli presented to the blind visual field in the absence of V1 activity in human subjects. To clarify the moment in time that visual information reaches extrastriate areas, by means of event-related potentials (ERPs) we tracked the temporal course of responses to complex visual stimuli (faces) presented in the blind field of a hemianopic patient. Stimulation of the normal visual field elicited a positive occipital deflection (P1) at 140 ms. A P1 response was also observed with stimulation of the blind field, although slightly delayed (20 ms) and reduced. Its topography and timing demonstrate that early neural activity for stationary stimuli takes place within extrastriate regions despite V1 denervation. PMID- 10670780 TI - Harmonic partials facilitate pitch discrimination in humans: electrophysiological and behavioral evidence. AB - The effect of the spectral tone structure on pre-attentive and attentive pitch discrimination was investigated. The mismatch negativity (MMN) component was recorded from reading subjects to pitch changes of identical magnitude in pure tones with only one sinusoidal frequency component and in spectrally rich tones with two additional harmonic partials. In a separate condition, subjects were asked to indicate detection of pitch change by a button press. The MMN was elicited with a larger amplitude and shorter latency by change in spectrally rich tones than by change in pure tones. Furthermore, the subjects' behavioral responses were more accurate for spectrally rich tones than for sinusoidal tones. Together these data indicate that pre-attentive and attentive pitch discrimination is facilitated with spectrally rich sounds in comparison to pure sinusoidal tones. PMID- 10670781 TI - Brain-derived neurotrophic factor in patients with frontotemporal dementia. AB - Brain-derived neurotrophic factor (BDNF) promotes survival and growth of various nerve cell populations during normal development and following different insults in the developing and adult brain. BDNF expression is reduced in Alzheimer disease, but little is known about BDNF expression in other types of dementia. Frontotemporal dementia (FTD) is a common cause of mental impairment in old age, which is characterized by neuron loss in the upper cortical layers mainly of the frontal and temporal cortex. BDNF protein expression has been examined by Western blotting and immunohistochemistry in the cerebral cortex of individuals affected by FTD. Examination of pathological samples (n = 8, mean age: 74.7 years; four men, four women) was conducted in parallel with corresponding samples from age matched controls (n = 8; mean age: 72.6 years; three men, five women). Post mortem delay was between 2 and 6 h. Preserved BDNF expression, as revealed by Western blotting, has been observed in the frontal and temporal cortices of patients with FTD. Furthermore, immunohistochemistry has disclosed maintained BDNF immunoreactivity in surviving neurons of the upper cellular layers, as well as in neurons of the inner cellular layers in FTD. These results show that FTD is not associated with a decay of BDNF in cortical neurons, and therefore, that BDNF is differentially regulated in diseases causing dementia. PMID- 10670782 TI - Characterization of [3H]mazindol binding sites in cultured monkey amniotic epithelial cells. AB - Our previous studies showed that monkey amniotic epithelial cells (MAEC) synthesize and release catecholamines and possess D1 and D2 dopamine (DA) receptors (Elwan, M.A., Ishii, T., Ono, F. and Sakuragawa, N., Evidence for the presence of dopamine D1 receptor mRNA and binding sites in monkey amniotic epithelial cells. Neurosci. Lett., 262 (1999) 9-12; Elwan, M.A., Ishii, T. and Sakuragawa, N., Detection of dopamine D2 receptor mRNA and binding sites in monkey amniotic epithelial cells. J. Neurosci. Res., 56 (1999) 316-322; Elwan, M.A., Thangavel, R., Ono, F. and Sakuragawa, N., Synthesis and release of catecholamines by cultured monkey amniotic epithelial cells. J. Neurosci. Res., 53 (1998) 107-113). In the present study we tested the presence of DA transporter (DAT) in MAEC using radioligand binding experiments. Saturation studies showed that [3H]mazindol binds to a high affinity site with K(D) and Bmax values of 7.85 +/- 1.25 nM and 123.22 +/- 18.34 fmol/mg protein, respectively. Competition studies indicated that selective DAT inhibitors are potent displacers of [3H]mazindol binding, compared to inhibitors of other types of transporters. The rank order of potency of the competing drugs is consistent with the pharmacology of DAT. These results provide, for the first time, clear evidence that MAEC natively possess DAT binding sites and suggest that MAEC may provide a potential primate cell model to study DA release and uptake processes and to explore new drugs active at this site. PMID- 10670783 TI - No evidence for long CAG/CTG repeats in families with spastic paraplegia linked to chromosome 2p21-24. AB - Autosomal dominant familial spastic paraplegia (AD-FSP) is a genetically heterogeneous, neurodegenerative disorder characterized by spasticity and progressive weakness in the lower limbs. Anticipation has been suggested to occur and an association between expanded CAG/CTG repeats and AD-FSP linked to the SPG4 locus (2p21-p24) has been described. In this study, 42 affected individuals from six SPG4 families were screened for expanded CAG/CTG repeats using the repeat expansion detection (RED) method. Large RED products (range 180-240 nucleotides) corresponding in size to repeats at the ERDA1 locus were detected in eight patients and at the CTG 18.1 locus in one patient. The large ERDA1 repeats did not segregate with the disorder within families. Mean age at onset and index of severity were not significantly different between patients with or without expanded RED products. Furthermore, no abnormal proteins were found by Western blot in 15 selected patient samples as compared with controls, using the 1C2 antibody, which detects long polyglutamine stretches. Thus, in contrast to previous reports, our study provides evidence against the hypothesis that a large translated CAG repeat expansion is the basis of SPG4. We propose that mechanisms other than large pathogenic CAG/CTG repeats may account for the disease in the SPG4 families tested here. PMID- 10670784 TI - Intraseptal infusions of 8-OH-DPAT in the rat impairs water-maze performances: effects on memory or anxiety? AB - In the rat, 5-HT1A receptors are found on medial septal cholinergic neurons. The effects of intraseptal infusions of the 5-HT1A receptor agonist 8-OH-DPAT (8 hydroxy-2-(di-n-propyl-amino)-tertralin) were assessed on reference memory performances in a water maze. Compared with vehicle infusions, 0.5 and 4 microg of 8-OH-DPAT significantly impaired (but did not prevent) acquisition of the task and probe-trial performances. The results suggest that activation of 5-TH1A receptors in the (medial) septal area impairs spatial learning, perhaps directly by reducing the hippocampal cholinergic tonus, or indirectly by an effect on anxiety. PMID- 10670785 TI - Effects of extracellular Ca2+ on membrane and seal resistance in patch-clamped rat thalamic and sensory ganglion neurons. AB - We studied the effects of [Ca2+]ext changes on seal resistance in patch-clamp experiments. Recordings were made on rat peripheral and thalamic neurons. Increasing [Ca2+]ext from 0.5 to 4.5 mM, reduced the ionic currents evoked at potentials from -100 to +50 mV, in cell-attached recordings, in all the neurons tested. The effect was greater at negative potentials. The change in seal conductance (deltaG) decreased with higher resistance seals and became very low over 1 Gohm (<0.5 nS). However, the ratio deltaG/G(0.5 Ca2+) rose from close to 0 up to 0.6, indicating that Ca2+ has a stronger effect when the microelectrode and the membrane are sealed more tightly. These findings suggest that changes in seal resistance may be misleading in experiments in which extracellular Ca2+ changes are used. PMID- 10670786 TI - Gender and age-related variation in adenylyl cyclase activity in the human prefrontal cortex, hippocampus and dorsal raphe nuclei. AB - The influence of gender and age on adenylyl cyclase activity was investigated, through a Dowex-alumina double step chromatographic procedure, in the prefrontal cortex, hippocampus and dorsal raphe nuclei obtained from autopsy cadavers. Results showed that forskolin-stimulated enzyme activity in raphe nuclei was greater in men than in women; a region-dependent rank order of basal, forskolin induced adenylyl cyclase activity and percentage forskolin-stimulation was observed in women only. Lastly, basal values correlated positively with forskolin stimulated adenylyl cyclase activity in all areas except the prefrontal cortex of the male subjects. Positive significant correlations were also found between both forskolin-stimulated enzyme activity and percentage forskolin stimulation and aging in the prefrontal cortex. Overall, the findings suggest that sex and/or age related differences in brain adenylyl cyclase vary from one cerebral region to the other. PMID- 10670787 TI - Target cells of apoptosis in the adult murine dentate gyrus and O4 immunoreactivity after ionizing radiation. AB - The occurrence of radiation-induced apoptosis and the determination of target cells were investigated by using the TdT-mediated dUTP-biotin nick end labeling assay and immunohistochemical analyses. The O4 immunoreactivity, an oligodendrocytes surface antigen, was also evaluated by using western blotting analysis. C57BL/6J adult female mice were subjected to single dose irradiation of 10 Gy. Eight hours after irradiation, the most significant increase of apoptotic cells was detected in the subgranular zone and the hilus of the dentate gyrus. The target cells of radiation-induced apoptosis are the subgranular progenitor cells and the oligodendrocytes in the hilus. The amount of the O4 immunoreactivity, a marker for premature oligodendrocytes, was unchanged until 8 h but enhanced after 12 h of irradiation. These results are the first to show the increase of the O4 immunoreactivity after irradiation and may be associated with the pathogenesis of radiation injury. PMID- 10670788 TI - Evidence that N-terminal fragments of nociceptin modulate nociceptin-induced scratching, biting and licking in mice. AB - The intrathecal (i.t.) injection of 3.0 fmol nociceptin (orphanin FQ) elicited scratching, biting and licking responses in mice. N-terminal fragments of nociceptin, nociceptin (1-7), nociceptin (1-9) and nociceptin (1-13), induced no characteristic behavioral response. When these N-terminal fragments of nociceptin were injected simultaneously with nociceptin, the behavioral response induced by nociceptin was reduced dose-dependently. Nociceptin (1-13) was much more potent than nociceptin (1-7) and nociceptin (1-9) and antagonized nociceptin-induced response at equimolar doses. No significant effects of the N-terminal fragments were observed against the scratching, biting and licking response elicited by i.t. administration of substance P or N-methyl-D-aspartate. These results suggest that N-terminal fragments formed endogenously in the spinal cord may have an antagonistic effect on nociceptin-induced behavioral responses. PMID- 10670789 TI - S-100 protein-immunoreactive structures in the brains of the elasmobranchs Scyliorhinus torazame and Mustelus manazo. AB - S-100 protein-immunoreactive structures were investigated in the brains of two species of elasmobranchs, Scyliorhinus torazame and Mustelus manazo. In both species, immunoreactivity for S-100 protein was seen widely in the brain. It was localized in astrocytes and tanycytes; superficial glial membrane, vascular endfeet, and radial fibers were recognized clearly. However, no immunoreactivity was found in the ependymal cells of non-tanycyte type, which were located in the choroidal plexus, subcommissural organ, and saccus vasculosus. No neuronal cells showed S-100 protein immunoreactivity. Immunoelectron microscopy revealed that the antigen was present diffusely in the cytoplasmic matrix of the cells and also associated with the plasma membrane, outer membrane of mitochondria, and microtubule-like components. PMID- 10670790 TI - Post-perfusion syndrome and disturbed microcirculation after cardiac surgery: the role of angiotensin-converting-enzyme inhibitors. AB - BACKGROUND: The sympathoadrenal and the renin-angiotensin system (RAS) are involved in blood pressure regulation. They are known to be activated during cardiac surgery. We investigated the influence of preoperative RAS-blockade using angiotensin-converting-enzyme inhibitors (ACEI) on hemodynamic variables and on the perioperative need for exogenous catecholamines. METHODS: 240 patients undergoing coronary artery bypass grafting (CABG) or valve surgery were divided into three matched groups (group A: pre- and postoperative ACEI; group B: ACEI only pre-, not postoperatively; group C: no ACEI). In these three groups we analyzed hemodynamic variables, the need for catecholamines and the incidence of a "post-perfusion syndrome" or systemic inflammatory response syndrome (SIRS) with impaired microcirculation. RESULTS: There were significant differences in the intra- and postoperative need for catecholamines in groups A and B compared to C (intraop. A: 35%, B: 35%, C: 15%; postop. A: 21.2%, B: 16.2%, C: 10%) (p < 0.05). In the ACEI groups (A and B) there were 9 patients with a postoperative SIRS, only 2 cases in group C. Furthermore 4 patients of group B suffered from disturbances of the intestinal microcirculation postoperatively. CONCLUSIONS: Long-term ACEI treatment before cardiac surgery raises the perioperative need for catecholamines. Patients with preoperative long-term use of ACEI who do not receive ACEI postoperatively face an increased risk of impaired microcirculation. The inhibition of angiotensin-II (AT II) generation causes the vasodilatatory effects of ACEI, and could be one reason for a post-perfusion syndrome or a SIRS. PMID- 10670791 TI - Oral administration of the dopamine prodrug docarpamine shortens need for drip infusion of dopamine in patients with low cardiac output syndrome after cardiac surgery. AB - BACKGROUND: Docarpamine (DOC) is a dopamine prodrug which can be orally administered. It has been found that oral docarpamine transforms into dopamine in vivo, and increases cardiac output and renal blood flow as effectively as intravenous dopamine. METHODS: We reviewed the records of 26 patients who had developed low cardiac output syndrome (LOS) after cardiac surgery and received docarpamine during the early postoperative course. Five patients discontinued docarpamine within 2 days due to arrhythmia. There were 3 hospital deaths. The remaining 18 patients were divided into two groups according to the timing of docarpamine administration. In group A docarpamine was administered during and after weaning from intravenous catecholamines, in group B only on demand after weaning from intravenous catecholamines. RESULTS: There were 12 patients in group A and 6 in group B, and the severity of LOS was relatively milder in group B than in group A. Stable hemodynamics and sufficient daily urinary output were maintained by oral administration of DOC in both groups just as well as by drip infusion of catecholamines. CONCLUSION: Sinse continuous drip infusion of catecholamine commonly slows recovery in LOS patients, it is considered that switching from drip infusion of catecholamines to oral DOC administration is safe and useful for earlier recovery in LOS patients after cardiac surgery. PMID- 10670792 TI - Quadrileaflet stentless mitral valve replacement. AB - BACKGROUND: The study evaluates clinical results and hemodynamic parameters one year after implantation of a stentless quadrileaflet mitral valve (QMV). METHODS: Since August 1997 28 patients received the QMV, patient age was 69 +/- 8 years; 13 had predominant mitral stenosis and 15 incompetence, preoperative NYHA functional class was III or IV and cardiac index 1.8 +/- 0.6 L/min/m2. RESULTS: Surgery was performed using a conventional (25) or a minimally invasive approach (3). 20 patients received a medium and 8 a large-size prosthesis, crossclamp time was 58 +/- 19 min. Additional procedures were myocardial revascularization in four, tricuspid repair in two, and left-atrial radiofrequency ablation to restore sinus rhythm in six patients. Perioperative mortality (1) was not valve-related. All other patients were discharged on time. At postoperative, 6-, and 12-months follow-up mean transvalvular pressure gradients were 4.2 +/- 1.5 / 4 +/- 0.9/ 3.8 +/- 1.4 mmHg and mitral valve orifice area index was 1.5 +/- 0.3 / 1.6 +/- 0.3 / 1.6 +/- 0.4, NYHA class was I or II. CONCLUSIONS: The QMV is well suited for mitral valve replacement. The anulo-ventricular continuity is preserved and the QMV function resembles native mitral valve function. If its performance is maintained in the long term the QMV may be the mitral prosthesis of choice. PMID- 10670793 TI - Perioperative factors influencing interleukin-10 release under cardiopulmonary bypass. AB - BACKGROUND: The cytokine response to cardiopulmonary bypass (CPB) is complex and can be modified. Among several mediators, the anti-inflammatory interleukin-10 (IL-10, 'cytokine-secretion inhibitory factor') is particularly interesting because of its ability to counteract pro-inflammatory cytokines triggering endothelial and leukocyte activation in the immediate immune response to CPB. On the other hand, during the delayed phase of the immune response, IL-10 may act as a promotor of immunodeficiency in complicated courses. Therefore, it is of interest to investigate special conditions of CPB that may influence the extent of perioperative release of IL-10. METHODS: We analyzed 20 continuously registered parameters during CPB, including an analysis of subgroups in the case of application of aprotinin or steroids. 30 consecutive adult patients with coronary artery disease (CAD) and normal left-ventricular function undergoing elective CABG were prospectively studied. Arterial blood was sampled perioperatively and levels of IL-10 were determined using ELISA tests. For analysis, the time point of maximum IL-10 release was selected (30 min after end of CPB). Simultaneously, CPB-registration protocols were analyzed concerning standard parameters. RESULTS: We could state an exponential relationship between IL-10 levels 30 min after end of CPB and the ischemia time (r = 0.76), duration of CPB (r = 0.73) and the extent of negative base excess (BE, r = 0.66) in all subgroups. An inverse relationship could be seen between IL-10 plasma levels and venous O2 saturation: low values for O2 saturation correlated with high IL-10 levels as did low mean arterial pressure (MAP). Hypothermia reduced IL-10 release (r = 0.80), whereas a long duration correlated with high IL-10 release (r = 0.67). In the case of longer duration of hypothermia, the protective effect vanished. CONCLUSIONS: The results show a significant rise for IL-10 early after starting CPB. Low values for venous O2 saturation and low MAP correlated with high IL-10 levels. A good correlation could be seen between IL-10 plasma levels and the duration of CPB, ischemia time, and negative base excess. Because of the ability of persisting IL-10 production to induce a higher incidence of septic complications, all actions for maintaining an optimum of perfusion and oxygenation play an important role. PMID- 10670794 TI - A new diagnostic procedure for assessing intracardiac flow disturbances in patients with heart valve disease. AB - BACKGROUND: Until now no diagnostic technique was available for the three dimensional (3D) study of intracardiac blood flow abnormalities in patients with heart valve disease. 3D color Doppler is a new diagnostic technique first developed at our institution. METHODS: The 3D reconstructions of the blood flow velocity data have been obtained from conventional multiplanar transesophageal or transthoracic Doppler echocardiographic examinations. We analyzed 111 reconstructions of color Doppler data obtained from 85 patients with different heart valve diseases who underwent intraoperative transesophageal echocardiography. Sixty-nine patients had a significant mitral regurgitation, 7 mitral stenosis, 9 aortic regurgitation, 12 aortic stenosis, 14 tricuspid regurgitation. Three patients had pulmonary regurgitation associated with mitral valve disease. RESULTS: 3D color Doppler disclosed the complex spatial spreading of the blood flow abnormalities caused by heart valve disease. New patterns of intracardiac blood flow disturbances could be observed and classified. CONCLUSIONS: This paper shows the first clinical applications of 3D color Doppler in patients with heart valve disease. The new insights derived from the 3D study of intracardiac blood flow dynamics revealed a great impact of this technique on the clinical management of patients with heart valve disease. PMID- 10670796 TI - Systemic blood pressure and cerebral blood flow velocity during carotid surgery. AB - BACKGROUND: To evaluate the effect of mean arterial blood pressure (MAP) on cerebral perfusion during carotid surgery, we investigated blood flow velocity in the middle cerebral (Vs,mca) using transcranial Doppler ultrasonography (TCD). METHODS: During carotid crossclamping, treatment included either phenylephrine induced hypertension without shunting (Group XC; n = 11) or insertion of a shunt (Group S; n = 12). RESULTS: Increasing MAP in Group XC before crossclamping (81 + /-13 mmHg to 107 +/- 12 mmHg) caused an increase of Vs,mca (59 +/- 17 cm/s to 75 +/- 20 cm/s; p < 0.001). During crossclamping without a shunt, Vs,mca was not dependent on MAP, and was reduced (mean 47 +/- 24 cm/s) in relation to preclamp values. In Group S, Vs,mca was always dependent on MAP and the preclamp velocity was maintained (before shunt: 75 +/- 26 cm/sec; during shunt: 79 +/- 30 cm/sec). CONCLUSIONS: Although we found an impaired cerebral autoregulation, Vs,mca was independent of MAP during carotid crossclamping. Thus, TCD measurements have to be interpreted with caution during crossclamping, and the effect of induced hypertension has to be confirmed with more invasive measures of cerebral blood flow. PMID- 10670795 TI - Cardiac surgery in patients with previous carcinoma of the breast and mediastinal irradiation: is the internal thoracic artery graft obsolete? AB - BACKGROUND: The increasing number of patients of more advanced age undergoing cardiac surgery means the number of those with previous curative (relapse free) mastectomy and irradiation of the chest is also increasing. A higher incidence of postoperative complications such as sternal infection in these patients is considered possible. Furthermore the question of whether mediastinal irradiation leads to a relevant internal thoracic artery (ITA) gaft damage remains unclear. In this context the benefit of arterial revascularization (CABG) using one or both ITAs is not sufficiently proven by data available from clinical studies. METHOD: 70 patients (49-85 years) with previous mastectomy or Hodgkin/non Hodgkin's disease and mediastinal irradiation underwent CABG (n = 59) or an aortic valve replacement (AVR, n = 11). 20 patients received bilateral internal thoracic artery grafts, 34 a single internal thoracic artery graft, and in 16 patients an internal thoracic artery was not used. Perioperative data and data concerning postoperative complications such as mortality, myocardial infarction, and sternal infection or refixation was gathered and compared with all other patients receiving CABG (n = 5102). An histological investigation of ITA segments was done in 12 patients. RESULTS: There was no significant enhancement of the perioperative risk in comparison with other patients of a corresponding age group. Internal thoracic artery damage induced by irradiation was not present. There was no increased incidence of sternal instability requiring refixation observed. CONCLUSION: In the patient cohort investigated there is in general no need for restrictive use of the ITA in CABG. PMID- 10670797 TI - A human heterotopic transplant on the beating heart: an interim report of a successful technique performed at Kocaeli University. AB - Heterotopic heart transplantation is still indicated in selected patients mainly with pulmonary vascular bed problems. Cardio-pulmonary bypass (CPB) has well known deleterious effects on the pulmonary vascular bed due to leukocyte sequestration as well as on the immune response of the patients. Also the negative effects of the cardioplegia on the native heart is another drawback of the classical heterotopic heart transplantation with the use of CPB. We want to present our new implantation technique for heterotopic heart transplantation without CPB, used successfully in our institution at the Kocaeli University on a patient with resistant pulmonary hypertension and dilated cardiomyopathy. PMID- 10670798 TI - Simultaneous coronary artery bypass grafting and transmyocardial laser revascularization through a small left thoracotomy. AB - We report a patient in whom coronary artery bypass grafting with the left internal mammary artery to the left anterior descending coronary artery and laser transmyocardial revascularization were simultaneously performed through a left small thoracotomy. The patient recovered uneventfully and 9 months following surgery he is free of angina and has increased effort tolerance. This case underlines the feasibility of combining these two minimally invasive procedures through the same approach in selected patients. PMID- 10670800 TI - Respiratory insufficiency caused by an aneurysm with multiple vascular lesions. AB - A 71-year-old woman, who presented tracheobronchial obstruction caused by a thoracic aortic aneurysm, was admitted to our institution. Although she had multiple cerebral infarctions, old myocardial infarction, bilateral iliofemoral atherosclerotic lesions with abdominal aortic aneurysm, and superior vena cava syndrome, aneurysmectomy was undertaken in order to rescue her from respiratory insufficiency. The operation successfully relieved her of exertional dyspnea and dysphagia. PMID- 10670799 TI - Successful streptokinase lysis of a right-atrial thrombus in a heart-transplant recipient. AB - This report describes a mobile right-atrial thrombus formation diagnosed by transthoracic echocardiography in a 22-year-old patient seven months after orthotopic heart transplantation. To prevent embolic complication, systemic streptokinase lysis was performed over 48 hours. Echocardiographic control examinations have shown that this therapy completely removed the thrombus. PMID- 10670801 TI - Tracheobronchial rupture after emergency intubation. AB - Iatrogenic injuries of the trachea are rare. We report a case of tracheobronchial rupture in a 77-year-old woman after emergency intubation. Early bronchoscopy showed a rupture of the posterior wall of the trachea into the right main bronchus with a total length of 9.5 cm. A right thoracotomy was performed and the tracheal rupture was managed successfully by primary suture. Surgical treatment and possible causes of this lesion are discussed with reference to the literature. PMID- 10670802 TI - A surgical management of aortic insufficiency concomitant with mediastinal well differentiated liposarcoma. AB - We present a rare case of a mediastinal liposarcoma concomitant with aortic insufficiency due to myxoid degeneration of the aortic valve. Because the patient's left ventricle was in moderate dilatation and a posterolateral thoracotomy combined with median sternotomy was required in order to perform a complete resection of a mediastinal liposarcoma, it was decided to carry out aortic valve surgery and tumor excision in one operation. PMID- 10670803 TI - Carbon dioxide insufflation aids video-assisted thoracic surgery in a young child. AB - A 3-year-old girl with pectus excavatum successfully underwent exploration using video-assisted thoracic surgery. A complete pericardial defect was identified. The lung was kept totally collapsed during the procedure using low-flow (1 L/min), low-pressure (7 mmHg) carbon dioxide insufflation. This technique is expected to be a safe adjunct to thoracoscopic procedures in infants and small children. PMID- 10670804 TI - A congenital defect of the pericardium. AB - Ten patients with congenital defects of the pericardium were treated in Departments of Cardiac Surgery, Silesian School of Medicine in Zabrze and Katowice between 1989 and 1998. There were eight children and two adults, eight males and two females. In each case the pericardial defect was discovered intraoperatively during surgery for congenital heart defect. There were no cases with clinical symptoms that could be clearly related to the defect of the pericardial sac. In the case of a child with a complete absence of the left pericardial wall the heart was significantly rotated contrary to the defect. The final outcome of the congenital heart defect surgery was satisfactory in each case. An abbreviated historical review of the diagnosis and treatment of the pericardial defects is presented with special attention placed on therapeutic management. Surgical correction of pericardial defects is concluded to be justified in patients with clinical symptoms. In most cases pericardial defects are discovered intraoperatively, but when they are large the said defects do not require any treatment. PMID- 10670805 TI - Current practice of peri- and postoperative antibiotic therapy in cardiac surgery in Germany. Working Group on Cardiothoracic Surgical Intensive Care Medicine of the German Society for Thoracic and Cardiovascular Surgery. AB - BACKGROUND: The increasing development of antimicrobial resistance of common bacterial pathogens presents one of the most significant challenges to clinical medicine, particularly intensive care medicine. One factor which has contributed to this development is the (over)use of antibiotic treatment. Therefore the objective of this study was to scrutinize the current practice of empiric antibiotic therapy in cardiac surgery in Germany for 1) perioperative prophylaxis and 2) postoperative therapy prior to the availability of susceptibility patterns for the infecting pathogen. METHODS: A questionnaire was sent to all centers performing cardiac surgery in Germany. Questions referred to drugs used as well as dosage, homogeneity and duration of antibiotic prophylaxis, time and/or reason for changing this regimen, drugs used for first-, second-, and third-line empiric postoperative antibiotic treatment, and homogeneity of antibiotic usage. RESULTS: All but 3 institutions (96.3%) answered. 1. Perioperative prophylaxis: All but 4 centers (94%) use first- (n = 32 = 43%) or second-generation cephalosporins (n = 38 = 51%) most commonly for 24 hours (n = 60 = 81%). Prophylaxis never exceeds 3 days. 74% of all institutions (n = 55) use the same antimicrobial agent for all cardiac procedures performed, while 26% (n = 19) change their regimen in selected patient groups, most commonly for heart transplantation. The entire prophylaxis is changed mainly according to susceptibility patterns (n = 63 = 85%), 7 centers (10%) change according to a fixed time schedule, while 4 institutions (5%) never change the antimicrobial drug. 2. Empiric postoperative therapy: A total of 29 different antibiotics out of 8 subclasses are used. No major differences between 1st-, 2nd-, and 3rd-line therapy could be detected, with the exception of a decreasing usage of beta-lactams (carbapenems excluded) from 60% in 1st-line to 23% in 3rd-line therapy and an increasing usage of glycopeptides from 5% in 1st line to 18% in 3rd-line therapy. 41 institutions (55%) use the same antibiotic regimen on the intensive care unit and the normal ward, 9 centers (12%) use the same drug for perioperative prophylaxis and postoperative therapy, and 12 institutions (16%) prescribe a combination therapy. CONCLUSIONS: Perioperative prophylaxis in cardiac surgery in Germany is performed on a relatively uniform basis and at low cost. The heterogeneity of antibiotic regimens for postoperative therapy may indicate the need for recommendations and/or guidelines for this type of treatment. The indications for the usage of reserve antibiotics, e.g. vancomycin, implying the possible risk of creating pathogens with untreatable resistance patterns, as well as strategies aimed at preventing the development of resistance should be the subject of further discussions. PMID- 10670806 TI - Kinetic study of the mass transfer of bovine serum albumin in anion-exchange chromatography. AB - A kinetic study was made on the mass transfer phenomena of bovine serum albumin (BSA) in two different anion-exchange columns (Resource-Q and TSK-GEL-DEAE-5PW). The analysis of the concentration dependence of the lumped mass transfer rate coefficient (km,L) provided the information about the kinetics of the several mass transfer processes in the columns and the anion exchangers, i.e., the axial dispersion, the fluid-to-particle mass transfer, the intraparticle diffusion, and the adsorption/desorption. In the Resource-Q column, the intraparticle diffusion had a dominant contribution to the band broadening compared with those of the other processes. The surface diffusion coefficient (Ds) of BSA showed a positive concentration dependence, by which the linear dependence of km,L on the BSA concentration seemed to be interpreted. On the other hand, in the TSK-GEL-DEAE 5PW column, the contribution of the adsorption/desorption was also important and almost same as that due to the intraparticle diffusion. There are some differences between the intrinsic properties of the mass transfer kinetics inside the two anion exchangers. It was likely that the positive concentration dependence of Ds was explained by the heterogeneous surface model. PMID- 10670807 TI - Synthesis of a silica-bonded bovine serum albumin s-triazine chiral stationary phase for high-performance liquid chromatographic resolution of enantiomers. AB - A novel method of synthesizing protein chiral stationary phase (protein-CSP) is proposed with 2,4,6-trichloro-1,3,5-triazine as the activator. The bovine serum albumin (BSA) based chiral columns (150 x 4.6 mm I.D.) were prepared successfully within 8 h. With tryptophan as the probe solute, it was observed that the BSA immobilized by this method had a better ability to distinguish enantiomers than that activated by glutaric dialdehyde. This may be due to the well-maintained BSA conformation and the larger amount of BSA immobilized on the silica gel. The BSA CSP prepared by this method was relatively stable under experimental conditions, and the resolution of 13 chiral compounds was achieved. The coupling reaction in this method is mild, reliable and reproducible; it is also suitable for the immobilization of various biopolymers in the preparation of bioreactor, biosensor and affinity chromatography columns. PMID- 10670808 TI - Comparison of the enantioseparation of racemic uridine analogs on Whelk-O 1 and ChiralPak-AD columns. AB - The commercially available, brush-type (S,S)-Whelk-O 1 chiral stationary phase (CSP) has been used to separate 10 racemates of structurally related uridine analogs, potentially anti-viral agents, under various mobile phase compositions, using various temperatures. The enantioseparation was evaluated by comparing the Whelk-O 1 column performance with that of ChiralPak-AD column, reported previously. The comparison involved the role of some distinctive structural features of the racemates, type and composition of the solvent modifiers, as well as effect of temperature on the chiral discrimination. Despite the fact that both columns separate almost all the uridine analogs, significant differences were observed in their chiral recognition, as revealed from their retention, selectivity, resolution and elution order. The chiral recognition processes, responsible for enantioseparation on the Whelk-O 1 column, were relatively more systematic and easier to manipulate than on ChiralPak-AD column. Enantioseparation on the latter are of more complex nature and frequently gave results that were contradictory to the expectations. On the other hand, the performance in the ChiralPak-AD column was superior to that of the Whelk-O 1 column. Limitations in column handling and maintenance (pressure and temperatures) as well as limited solvent choice lead to the preference of the Whelk-O 1 column, in spite of its lower (but adequate) performance. PMID- 10670809 TI - Tandem solid-phase extraction of atrazine ozonation products in water. AB - The preconcentration of aqueous solutions containing atrazine degradation products was investigated using solid-phase extraction on octadecyl and cation exchanger silica phases. The retention and elution steps were studied and evaluated separately in order to define and optimize the critical experimental parameters involved. A strategy which combines sequentially both phases is proposed to fractionate compounds into two groups of increasing polarities: firstly, the native pesticide, hydroxyatrazine and most chlorotriazines on octadecyl support, and secondly monodealkylated hydroxytriazines, ammeline and ammelide on cation-exchanger. This tandem procedure was successfully applied for analysing and quantifying atrazine ozonation products and its efficiency demonstrated using [U-ring 14C]-labelled atrazine experiments. PMID- 10670810 TI - New operational modes for multidimensional and comprehensive gas chromatography by using cryogenic modulation. AB - Historically, hardware and method-related concerns have limited the use of multidimensional gas chromatography in the routine laboratory. This paper presents a new approach that offers the potential to significantly alter the manner in which multidimensional gas chromatography is conducted, based on the use of a modulated cryogenic trap which can be moved longitudinally along the column. Two columns are directly coupled, and no switching valves are used. It is demonstrated that a heartcut section can be cryofocused and zone-compressed, and then rapidly remobilized at the prevailing column oven temperature without any supplementary heating. A short second dimension column is used, giving fast second dimension analysis. This allows a large number of heartcuts to be programmed for any one analysis. The 'ultimate' manifestation of multidimensional gas chromatography is the comprehensive GC technique (GC X GC). This is now simply effected by performing very rapid heartcuts at intervals on the order of 1/5th of the peak width of primary dimension peaks, and requires that the second dimension be able to complete the analysis of each collected zone on a similar timeframe. This paper uses a semi-volatile aromatic mixture to demonstrate these selected operational modes, that can be achieved with the longitudinal modulation method. The flexibility that arises from this approach is shown by the ability to swap between selected whole-peak enhancement and comprehensive modes during the one analytical run. The increased sensitivity that follows from peak compression is a further advantage, which would be beneficial for trace analysis. PMID- 10670811 TI - Enantiomer separation of polychlorinated biphenyl atropisomers and polychlorinated biphenyl retention behavior on modified cyclodextrin capillary gas chromatography columns. AB - Seven commercially-available chiral capillary gas chromatography columns containing modified cyclodextrins were evaluated for their ability to separate enantiomers of the 19 stable chiral polychlorinated biphenyl (PCB) atropisomers, and for their ability to separate these enantiomers from achiral congeners, necessary for trace environmental analysis of chiral PCBs. The enantiomers of each of the 19 chiral PCBs were at least partially separated on one or more of these columns. Enantiomeric ratios of eleven atropisomers could also be quantified on six columns as they did not coelute with any other congener containing the same number of chlorine atoms, so could be quantified using gas chromatography-mass spectrometry. Analysis of a lake sediment heavily contaminated with PCBs showed enantioselective occurrence of PCB 91, proof positive of enantioselective in situ reductive dechlorination at the sampling site. PMID- 10670812 TI - Gas chromatographic quantification of amino acid enantiomers in food matrices by their N(O,S)-ethoxycarbonyl heptafluorobutyl ester derivatives. AB - Several amino acid enantiomer derivatives were prepared with different chloroformates and analysed by gas chromatography (GC) on a Chirasil-L-Val GC column, at a temperature below 200 degrees C. Among them the N(O,S) ethoxycarbonyl heptafluorobutyl esters presented the best compromise between short retention times, high yield responses and good resolution for almost all the tested amino acids. These derivatives proved to be suited for quantification of amino acids in aqueous media, with L-p-chlorophenylalanine as internal standard. The developed procedure was applied to several food samples for determination of their free amino acid profiles. PMID- 10670813 TI - Selective extraction of hydrocarbons, phosphonates and phosphonic acids from soils by successive supercritical fluid and pressurized liquid extractions. AB - Hydrocarbons, dialkyl alkylphosphonates and alkyl alkylphosphonic acids are selectively extracted from spiked soils by successive implementation of supercritical carbon dioxide, supercritical methanol-modified carbon dioxide and pressurized water. More than 95% of hydrocarbons are extracted during the first step (pure supercritical carbon dioxide extraction) whereas no organophosphorus compound is evidenced in this first extract. A quantitative extraction of phosphonates is achieved during the second step (methanol-modified supercritical carbon dioxide extraction). Polar phosphonic acids are extracted during a third step (pressurized water extraction) and analyzed by gas chromatography under methylated derivatives (diazomethane derivatization). Global recoveries for these compounds are close to 80%, a loss of about 20% occurring during the derivatization process (co-evaporation with solvent). The developed selective extraction method was successfully applied to a soil sample during an international collaborative exercise. PMID- 10670814 TI - Study of dead volume measurement in packed subcritical fluid chromatography with ODS columns and carbon dioxide-modifier mobile phases. AB - Studies were done for providing a simple, rapid and reliable procedure of void volume measurement in packed subcritical fluid chromatography (pSubFC), with CO2 modifier mobile phases containing high modifier amounts. Methods used in RPLC with ODS columns were applied in pSubFC: gravimetric, homologous series linearisation and unretained marker injection. Results lead us to propose the method of marker injection to determine the void volume in pSubFC. Acetonitrile was chosen as the void volume marker among six tested markers. Furthermore, void volume variations vs. the modifier volume (from 5 to 45%) were studied for nine organic modifiers. The void volume variations were related both to adsorption desorption phenomena between the mobile phase and the stationary phase and to mobile phase density changes. These variations allowed the classification of the modifiers into four groups on the basis of the molecular interactions. PMID- 10670815 TI - Micellar electrokinetic chromatography for the analysis of D-amygdalin and its epimer in apricot kernel. AB - We have developed a simple, rapid and reproducible method for the determination of D-amygdalin and its epimer by using micellar electrokinetic chromatography (MEKC). Separation of D-amygdalin was performed in a 20 mM sodium borate buffer (pH 8.5) containing 300 mM sodium dodecyl sulfate using a bare fused-silica capillary. The eluates were monitored by the absorbance at 210 nm. The applied electric field was 278 V/cm, and the time needed for the separation of D amygdalin did not exceed 6 min. The calibration curve for D-amygdalin showed excellent linearity in the concentration range of 5-500 microg/ml. The migration time and the corrected peak area show relative standard deviations (n=6) of 0.86% and 1.48%, respectively. The limit of detection (S/N=3) for D-amygdalin was 2 microg/ml. Under acidic and neutral conditions, amygdalin exists only as the D form; however, under basic conditions, it shows both the D- and L-forms with a concentration ratio of 1:1.3 (D-amygdalin/L-amygdalin). Results of HPLC, UV-Vis spectrophotometry, and mass spectrometry reconfirmed the identification of D amygdalin and its epimer. The number of theoretical plates of D-amygdalin is about 100,000 in MEKC, which is significantly higher than approximately 8,000 of HPLC. This method has been successfully applied to the determination of amygdalin epimers in various apricot kernel extracts and pharmaceutical products. PMID- 10670816 TI - Comparative studies on the analysis of glycosylation heterogeneity of sialic acid containing glycoproteins using capillary electrophoresis. AB - Comparative studies concerning glycoform analysis of sialoglycoproteins by capillary electrophoresis were performed using a few separation modes hitherto reported. Glycoprotein samples examined in the present study were successfully separated to their respective glycoforms using surface-modified capillaries commercially available for capillary gas chromatography in the running buffer near their isoelectric points. The analysis times were less than 50 min and reproducibilities in migration times were excellent (less than 2.0% RSD for both run-to-run and day-to-day analyses). We present a method for the glycoform analysis of alpha1-acid glycoprotein in sera by simple pre-treatment as an application. The present technique will become one of the general methods for the evaluation of glycosylation heterogeneity of commercially available glycoprotein drugs. PMID- 10670817 TI - Analysis of coptisine, berberine and palmatine in adulterated Chinese medicine by capillary electrophoresis-electrospray ion trap mass spectrometry. AB - Chinese medicine preparations contaminated with coptisine, berberine and palmatine were studied by capillary electrophoresis-electrospray ion trap mass spectrometry. The dubious adulterants were identified by their retention times, molecular ions and specific fragment ions produced from collision induced dissociation. The results showed that, in comparison with CE-UV and capillary electrophoresis-electrospray mass spectrometry (CE-ESI-MS), more reliable identification could be achieved with CE-ESI-MS-MS using ion trap mass spectrometry. PMID- 10670818 TI - Chromatographic separation of 3,4-difluorophenylacetic acid and its positional isomers using five different techniques. AB - The separation of five positional isomers from 3,4-difluorophenylacetic acid was investigated using normal- and reversed-phase high-performance liquid chromatography, capillary zone electrophoresis, gas chromatography and supercritical fluid chromatography. Operating parameters of each technique, such as temperature, type of stationary phase, mobile phase pH, ionic strength, organic modifiers and additives were varied in order to elucidate the separation mechanisms. Based on the advantages and disadvantages of each methodology, a simple and practical RPLC method was selected. The method was validated in terms of linearity, limit of detection, accuracy, recovery, ruggedness and precision. PMID- 10670819 TI - Capillary electrophoretic determination of sanguinarine and chelerythrine in plant extracts and pharmaceutical preparations. AB - Capillary electrophoresis was employed to determine the principal quaternary benzo[c]phenanthridine alkaloids, sanguinarine and chelerythrine, in two plant extracts and one oral hygiene product. Phosphate-Tris buffer of pH 2.5 was used as a background electrolyte, limits of detection were 3 micromol/l(-1) (sanguinarine) and 2.4 micromol,l(-1) (chelerythrine) using UV detection at 270 nm. The method, which correlated well with HPLC, is suitable for serial determination of sanguinarine and chelerythrine in plant products and pharmaceuticals. PMID- 10670820 TI - In vitro action of a combination of selected antimicrobial agents and chondroitin sulfate. AB - Chondroitin sulfate (CS), a highly anionic polymer and the most predominant sulfated glycosaminoglycan in connective tissues, was investigated regarding to its interaction with cationic disinfectants, which are used as antiinfectives in humans. Combinations of cetylpyridiniumchloride (CPC), chlorhexidine (CHex), and polyhexamethylene biguanide (PHMB) with CS, respectively, were prepared and the resulting microbicidal activity of the mixtures was tested in the quantitative suspension test without organic matter. Polyvidone-iodine and Ringer's solution were used as controls. Even precipitated, the resulting test combinations behave differently against Staphylococcus aureus, Enterococcus faecium, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans. CPC/CS demonstrated only microbicidal activity against Gram-positive bacteria, and CHex/CS was more active against Gram-negative bacteria and C. albicans. PHMB/CS, especially in combination with CS-A, only revealed an antimicrobial effect against P. aeruginosa after 60 min action. The interaction of cationic disinfectants with CS showed depending on the investigated microorganism a more or less controlled sustained release manner of the microbicidal agent from the precipitated complex, with the only exception of PHMB in combination with CS-C, which is completely neutralized. Polyvidone-iodine and Ringer's solution were not affected by CS. PMID- 10670821 TI - Potentiation of thioacetamide hepatotoxicity by phenobarbital pretreatment in rats. Inducibility of FAD monooxygenase system and age effect. AB - The ability of phenobarbital to induce the expression and activity of microsomal drug monooxygenases in the liver presents one of the most important issues in the field of chemical interactions and in the toxicity of xenobiotics. The model of rat liver injury induced by a single dose of thioacetamide (500 mg/kg intraperitoneally) was used to study the effect of phenobarbital (80 mg/kg/day intraperitoneally) for 5 days prior to thioacetamide. Serum parameters of liver injury such as aspartate aminotransferase activity, gamma-glutamyl transferase activity and the total bilirubin levels, as well as the activities of hepatic FAD and cytochrome P450 microsomal monooxygenases, were assayed in 2- and 12-month old rats. Samples of blood and liver were obtained from controls (injected at 0 h with 0.5 ml of 0.9% NaCl) and at 12, 24, 48, 72 and 96 h of thioacetamide intoxication either to non-treated or phenobarbital pretreated rats. Potentiation of thioacetamide hepatotoxicity by phenobarbital pretreatment was demonstrated at morphological level, and by significant increases in the activities of serum aspartate aminotransferase and gamma-glutamyl transferase, and in the levels of total bilirubin. The extent of potentiation of thioacetamide-induced liver injury by phenobarbital pretreatment was similar in both age groups. Microsomal FAD monooxygenase activity, the enzyme responsible for thioacetamide biotransformation, was significantly enhanced (twofold) by phenobarbital pretreatment, and also underwent a further increase following thioacetamide, preceding the peak of necrosis. Cytochrome P450 monooxygenases were induced by phenobarbital pretreatment more than sixfold, and sharply decreased when phenobarbital was withdrawn and thioacetamide administered, showing at 48 h intoxication values close to basal. Phenobarbital pretreatment potentiated thioacetamide necrogenicity, and this potentiation was parallel to the induction of the microsomal FAD monooxygenase system, both by phenobarbital and by thioacetamide itself. The extent of thioacetamide-induced liver injury was significantly higher in 12-month-old rats, but the effect of phenobarbital pretreatment was similar in both age groups. PMID- 10670822 TI - Conjugation of 1-naphthol in primary cell cultures of rat ovarian cells. AB - The present study concerns conjugation of 1-naphthol in primary cultures of rat ovarian cells. Two phase II enzymes catalyzing conjugation, i.e. phenol sulfotransferase (P-SULT) and phenol UDP-glucuronosyltransferase (P-UGT), were measured using 1-naphthol as substrate. The rates of conjugation by the different cell types of the rat ovary were the same at low concentrations and short incubation times. However, after 20 h of incubation the rate of conjugation in cells isolated from ovaries enriched in corpora lutea (CL) exceeded the rate in cells isolated from ovaries enriched in preovulatory follicles. In addition, when the granulosa cells were removed from the preovulatory follicles, the rate of conjugation was 1.7-fold higher, i.e. in the theca/stroma cells. When the cells were incubated with 1-[14C]naphthol and conjugates were subsequently separated by thin-layer chromatography, naphthyl glucuronide was the only conjugate observed. Pentachlorophenol (PCP), a commonly used inhibitor of P-SULT, inhibited 1 naphthol conjugation 50% in cell cultures, as well as in microsomal preparations. alpha-Naphthoflavone (ANF) and ellipticine (ELP), both cytochrome P450 (CYP) inhibitors, affected the conjugation of 1-naphthol in different ways; ANF did not affect P-UGT activity in microsomal preparations, but inhibited 1-naphthol conjugation in cell cultures by as much as 90%. On the other hand, ELP inhibited the conjugation of 1-naphthol up to 99% in the cell cultures, but only 75% in microsomal fractions. Testosterone (TST) and estradiol inhibited this activity approximately equal 50% in both of these experimental systems. Clomiphene citrate (CLF), a drug used to induce ovulation and demonstrating both estrogenic and antiestrogenic effects, did not influence the conjugation of 1-naphthol significantly in the cell cultures. The present findings demonstrate that P-UGT is by far the major enzyme conjugating 1-naphthol in the rat ovary and that commonly used inhibitors of P-SULT and CYPs also inhibit P-UGT activity, either directly or via other mechanisms. PMID- 10670823 TI - Induction by perfluorinated fatty acids with different carbon chain length of peroxisomal beta-oxidation in the liver of rats. AB - The potency of the induction of peroxisomal beta-oxidation was compared between perfluorinated fatty acids (PFCAs) with different carbon chain lengths in the liver of male and female rats. In male rats, perfluoroheptanoic acid (PFHA) has little effect, although perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA) potentially induced the activity. By contrast, PFHA and PFOA did not induce the activity of peroxisomal beta-oxidation in the liver of female rats while PFNA and PFDA effectively induced the activity. The induction of the activity by these PFCAs was in a dose-dependent manner, and there is a highly significant correlation between the induction and hepatic concentrations of PFCAs in the liver regardless of their carbon chain lengths. These results strongly suggest that the difference in their chemical structure is not the cause of the difference in the potency of the induction. Hepatic concentrations of PFOA and PFNA was markedly higher in male compared with female rats. Castration of male rats reduced the concentration of PFNA in the liver and treatment with testosterone entirely restored the reduction. In contrast to the results obtained from the in vivo experiments, the activity of peroxisomal beta oxidation was induced by PFDA and PFOA to the same extent in cultured hepatocytes prepared from both male and female rats. These results, taken together, indicate that difference in accumulation between PFCAs in the liver was responsible for the different potency of the induction of peroxisomal beta-oxidation between PFCAs with different carbon chain lengths and between sexes. PMID- 10670824 TI - Induction of rat hepatic drug metabolizing enzymes by dimethylcyclosiloxanes. AB - Low molecular weight dimethylcyclosiloxanes (DMCS) are important precursors in the synthesis of polydimethysiloxane polymers widely used in industry, and in medical and personal care products. The objective of this study was to characterize the ability of two DMCS, octamethylcyclosiloxane (D4) and decamethylcyclopentasiloxane (D5) to induce drug metabolizing enzymes in rats. Male and female Sprague-Dawley rats were administered 1, 5, 20, or 100 mg/kg D4 or D5 in corn oil daily by gavage for 4 days. Changes in the levels of activity and/or immunoreactivity of CYP1A1/2, CYP2B1/2, CYP3A1/2 and NADPH cytochrome P450 reductase in liver microsomes were examined. Significant increases were observed in the liver to body weight ratio in female rats administered either D4 or D5 at doses > or = 20 mg/kg. Increases in the liver to body weight ratio were observed in male rats treated with > or = 100 mg/kg D5 but not with D4. Relatively large increases in CYP2B1/2 enzymatic activity and immunoreactive protein were observed with increasing concentrations of both D4 and D5. Significant increases in 7 pentoxyresorufin O-depentylase (PROD) activity were also detected in male and female rats given D4 at doses > or = 5 mg/kg. D5 increased PROD activity in male rats at doses > or = 20 mg/kg and in female rats at doses > or = 5 mg/kg. 7 Ethoxyresorufin O-deethylase (EROD) activity was increased in both male and female rats receiving > or = 20 mg/kg D4 or > or = 5 mg/kg D5; however, no changes were detected in CYP1A1/2 immunoreactive protein in rats of either sex. D4 and D5 caused significant increases in CYP3A1/2 immunoreactive protein in only male rats treated with 100 mg/kg of either compound. However, significant increases were detected in CYP3A1/2 immunoreactive protein in female rats at D4 doses > or = 20 mg/kg and D5 doses > or = 5 mg/kg. Induction of NADPH cytochrome P-450 reductase immunoreactive protein was observed with D4 in female rats and in both male and female rats with D5. Induction of CYP2B/1/2, CYP3A1/2 and NADPH cytochrome P450 reductase was observed in rats treated with 50 mg/kg phenobarbital by intraperitoneal injection. Maximal CYP2B induction detected with D4 was approximately 50% of the increase observed with phenobarbital. In summary, D4 and D5 induced CYP2B1/2 in adult rat liver in a manner similar to that observed with phenobarbital; however, differences were observed between D4 and D5 in their ability to induce CYP3A1/2 and NADPH cytochrome P450 reductase. Female rats were more sensitive to the inductive properties of low doses of both DMCS than male rats whereas male rats were more responsive to phenobarbital induction. PMID- 10670825 TI - Renal production of thromboxane and prostaglandins in a rat model of type 2 diabetes. AB - In an investigation of the involvement of prostanoids in the pathogenesis of nephropathy in type 2 diabetes, we repeatedly measured the urinary excretion of prostanoids in both diabetic and healthy rats as the rats aged. Seven rats of the Otsuka Long-Evans Tokushima Fatty strain were used as rats with a model of type 2 diabetes and seven rats of the Long-Evans Tokushima Otsuka strain were used as rats without diabetes. Thromboxane (TX) B2 and 6-keto-prostaglandin (PG) F1alpha, the amounts of which reflect renal production of TXA2 and PGI2, respectively, and PGE2 in urine collected in metabolic cages were assayed when rats were 14, 30, 46, and 54 weeks old. Plasma glucose and urinary protein excretion also were measured periodically. The mean plasma glucose concentration of the diabetic rats was higher than that of the healthy rats throughout the study. At 30 weeks and later, urinary protein excretion by the diabetic rats was greater than that of the healthy rats, and it increased with age. Urinary excretion of TXB2 by the diabetic rats was higher than that of the healthy rats at 14 weeks (52.4+/-23.5 vs. 27.0+/-2.6 ng/day; mean +/- SD, P = .015) and the difference continued to the end of the experiment. Urinary excretion of 6-keto-PGF1alpha by the diabetic rats was high at 14 weeks (52.3+/-12.8 vs. 26.9+/-4.6 ng/day; mean +/- SD, P<.001) but decreased with age and was the same as that of the healthy rats at 54 weeks. The urinary excretion of PGE2 by the two groups of rats was not significantly different. These results suggest that altered renal production of TXA2 and PGI2 is involved in the pathogenesis of diabetic nephropathy in rats with type 2 diabetes. PMID- 10670826 TI - (N-stearyl, norleucine17) VIP hybrid inhibits the growth of pancreatic cancer cell lines. AB - The effects vasoactive intestinal peptide (VIP) antagonists were investigated on pancreatic cancer cell lines. (N-Stearyl, Norleucine17) VIP hybrid ((SN)VIPhyb) inhibited 125I-VIP binding to human Capan-2 cells with an IC50 value of 0.01 microM whereas VIP hybrid had an IC50 value of 0.2 microM. By RT-PCR and Northern blot, VPAC1 receptor mRNA was detected in CAPAN-2 cells. One microM (SN)VIPhyb and 10 microM VIPhyb inhibited the ability of 30 nM VIP to elevate cyclic AMP and increase c-fos mRNA. (SN)VIPhyb, 1 microM inhibited the clonal growth of CAPAN-2 cells in vitro. In vivo, (SN)VIPhyb (10 microg/day s.c.) inhibited CAPAN-2 xenograft growth in nude mice. These results indicate that (SN)VIPhyb is a pancreatic cancer VPAC receptor antagonist. PMID- 10670827 TI - Somatosensory evoked potential, neurological examination and magnetic resonance imaging for assessment of cervical spinal cord decompression. AB - The present study was designed to determine the relationship between neurological testing, anatomical imaging, and electrophysiological monitoring for assessing outcome of cervical spinal cord decompression. We prospectively studied 28 consecutive patients (age 39-76 yr) who were subjected to presurgical-(1-3 wk) and postsurgical (3-4 mo) neurological examination and recording of the median nerve somatosensory evoked potential (SEP). In 13 patients, magnetic resonance imaging (MRI) was also performed. Changes in neurological function, SEP and MRI were evaluated and graded as (1) improvement,(2) no change or (3) deterioration. Neurological outcome (NO) was based on changes in motor grade strength, sensory, reflexes and gait. The SEP outcome was based on changes in latency and disappearance of SEP waveform components whereas MRI evaluation was based on changes in spinal cord and canal diameters. Significance of association between NO, SEP and MRI was determined by Pearson's Chi-Square statistic (P<.05). The SEP improved in 71% (20/28) and deteriorated in 28% (8/28) of the subjects. An association between SEP changes and NO was found in 82% (23/28) of the subjects (P = .0038). Decompression increased the spinal canal diameter in 92% (12/13), and the spinal cord diameter in 38% (5/13) of the subjects. An association between NO, or SEP and MRI was not detected. Changes in median nerve SEP latency appear to be predictive of the neurological status of patients subjected to cervical spinal cord decompression. Postoperative increments in SEP latency or disappearance of the SEP waves were indicative of poor outcome after surgical decompression of the cervical spinal cord. PMID- 10670828 TI - Differential effects of hemorrhage and LPS on tissue TNF-alpha, IL-1 and associate neuro-hormonal and opioid alterations. AB - LPS administration and hemorrhage are frequently used models for the in vivo study of the stress response. Both challenges stimulate cytokine production as well as activate opiate and neuro-endocrine pathways; which in turn modulate the inflammatory process. Differences in the magnitude and tissue specificity of the proinflammatory cytokine and neuro-hormonal responses to these stressors are not well established. We contrasted the tissue specificity and magnitude of the increase in circulating and tissue cytokine (TNF-alpha, IL-1alpha and IL-1beta) content in response to either fixed-pressure hemorrhage (approximately 40 mm Hg) followed by fluid resuscitation (HEM) or lipopolysaccharide (LPS; 100 microg/100 g BW) administration. LPS and HEM elevated circulating levels of TNF-alpha, while neither stress altered circulating IL-1-alpha and IL-beta. LPS-induced increases in TNF-alpha content were greater than those elicited by HEM in all tissues studied except for the lung, where both stressors produced similar increases. Tissue (lung, spleen and heart) content of IL-1alpha was increased by HEM but was not affected by LPS. Tissue (lung, spleen, and heart) content of IL-1beta was increased by LPS but was not affected by HEM. HEM produced greater increases than LPS in epinephrine (16- vs. 4-fold) and norepinephrine (4-fold vs. 60%) levels and similar elevations in beta-endorphin. LPS produced greater elevation in corticosterone levels (2-fold) than HEM (50%). These results suggest differential tissue cytokine modulation to HEM and LPS, both with respect to target tissue and cytokine type. The hormonal milieu to HEM is characterized by marked catecholaminergic and moderate glucocorticoid while that of LPS is characterized by marked glucocorticoid with moderate catecholaminergic influence. PMID- 10670829 TI - Chinese green tea lowers cholesterol level through an increase in fecal lipid excretion. AB - Lung Chen Tea, a Chinese green tea, has been found to lower serum and liver cholesterol. In this study, its dose response and mechanisms of action on cholesterol lowering in diet-induced hypercholesterolemic Sprague-Dawley rats were investigated. The activities of three major lipid metabolizing enzymes, 3 hydroxy-3-methylglutaryl-coenzyme A (HMG-Co A) reductase, cholesterol 7alpha hydroxylase and fatty acid synthase (FAS), as well as fecal excretion of bile acids and cholesterol were examined. Lung Chen Tea administration for eight weeks significantly lowered the serum cholesterol in the 2% and 4% groups. The activities of the three enzymes were not affected by Lung Chen Tea, but the fecal bile acids and cholesterol excretions were significantly increased. These results demonstrated that Lung Chen Tea lowered plasma cholesterol by increasing fecal bile acids and cholesterol excretion. Further investigation is required to evaluate the exact mechanisms of action of Lung Chen Tea. PMID- 10670830 TI - Augmented platelet calcium uptake in response to serotonin stimulation in patients with major depression measured using Mn2+ influx and 45Ca2+ uptake. AB - There is an augmented platelet intracellular calcium response to serotonin stimulation in major depression. The role that calcium influx has in this process is not known. The objective of this study was to determine platelet calcium influx in response to serotonin by two methods, Mn2+ influx and 45Ca2+ uptake, in order to observe if the uptake response to serotonin was augmented in major depression by comparing the response to normal controls. The use of the two methods of calcium influx showed that serotonin stimulates calcium uptake into platelets. Furthermore, patients with major depression have significantly augmented platelet calcium uptake in response to serotonin. The interesting finding was that calcium uptake into platelets is biphasic, occurring immediately and after five minutes. These results may support the two pool model for calcium oscillations within cells whereby extracellular calcium is needed for intracellular calcium release, and for replenishment of depleted stores once intracellular calcium is released. PMID- 10670831 TI - Antinociceptive effect of (S)-N-desmethyl trimebutine against a mechanical stimulus in a rat model of peripheral neuropathy. AB - Trimebutine (2-dimethylamino-2-phenylbutyl 3,4,5-trimethoxybenzoate, hydrogen maleate) relieves abdominal pain in humans. In the present study, the antinociceptive action of systemic (S)-N-desmethyl trimebutine, a stereoisomer of N-monodesmethyl trimebutine, the main metabolite of trimebutine in humans, was studied in a rat model of neuropathic pain produced by chronic constriction injury to the sciatic nerve. Mechanical (vocalization threshold to hindpaw pressure) stimulus was used. Experiments were performed two weeks after surgery when the pain-related behaviour has fully developed. (S)-N-desmethyl trimebutine (1, 3, 10 mg/kg s.c.) produced dose-dependent antinociceptive effects on the nerve-injured and the contralateral hindpaw. The effect of the lowest dose (1 mg/kg s.c.) of (S)-N-desmethyl trimebutine on the nerve-injured paw was equal to that seen after a ten time stronger dose on the contralateral paw. The effect of (S)-N-desmethyl trimebutine (1 mg/kg) was not naloxone reversible. The results suggest that systemic (S)-N-desmethyl trimebutine may be useful in the treatment of some aspects of neuropathic pain. PMID- 10670832 TI - Effects of ouabain and flecainide on CO(2)-induced slowly adapting pulmonary stretch receptor inhibition in the rabbit. AB - The inhibitory effect of CO2 on slowly adapting pulmonary stretch receptors (SARs) was examined before and after administration of ouabain, a Na+-K+ ATPase inhibitor, and flecainide, a Na+ channel blocker. The experiments were performed in anesthetized, artificially ventilated rabbits after vagus nerve section. CO2 inhalation (maximal tracheal CO2 concentration ranging from 9.2 % to 10.4%) for about 60 sec decreased the receptor activity during both inflation and deflation. The magnitude of decreased SAR activity during deflation was greater than that seen during inflation. Administration of ouabain (25 microg/kg) initially stimulated SAR activities during inflation and deflation, and after 20 min, the SAR response was still kept excitatory in both inflation and deflation phases. Under these conditions, CO2 inhalation inhibited SAR activities during inflation and deflation. Flecainide treatment (3 mg/kg) that abolished veratridine (30 microg/kg)-induced SAR excitation had no significant effect on the inhibitory responses of SAR activity to CO2. These results suggest that the inhibitory effect of CO2 occurs when ouabain results in intracellular Na+ concentration ([Na+]i) increases in the SAR endings, and that CO2-induced SAR inhibition may not be related to the reduction of influx of Na+ through voltage-gated Na+ channels. PMID- 10670833 TI - Acute orexin effects on insulin secretion in the rat: in vivo and in vitro studies. AB - Orexin-A and orexin-B are members of a family of newly described orexigenic hypothalamic neuropeptides. Scanty data are available suggesting the involvement of orexins in regulation of the secretion of pituitary hormones and in control of energy homeostasis. Present studies aimed to explain whether orexins affect blood insulin concentration and insulin secretion in the rat. To check this possibility, adult female rats were subcutaneously injected with different doses (1 or 2 nmol) of orexin-A or orexin-B. A bolus administration of orexin-A resulted in an increase in blood insulin (up to min 120) and glucose (60 min after injection) concentration. The higher dose of orexin-B, on the other hand, exerted effect on insulin secretion only at min 60 of experiment and neither doses changed blood glucose level. Only orexin-A stimulated insulin secretion in an in vitro perfusion system of the rat pancreas preparation, while orexin-B was less effective. The results demonstrate that orexins belong to a group of neuropeptides influencing insulin secretion and acting directly on the pancreas. Direct, at least partial, effect of orexin on insulin secretion may be connected with the regulation of metabolism by this peptide. PMID- 10670834 TI - Oxidized LDL from subjects with different dietary habits modifies atherogenic processes in endothelial and smooth muscle cells. AB - Low-density lipoprotein (LDL) activates a number of processes involved in atherogenesis and vasoconstriction. Evidence suggests that oxidation increases the atherogenicity of LDL. We investigated the effects of oxidized LDL (ox-LDL) on cytotoxicity, prostacyclin (PGI2), and cyclic guanosine-3',5'-monophosphate (cGMP) production in rat vascular smooth muscle cell (VSMC) and rat heart endothelial cell (EC) culture, as well as EC- and VSMC-mediated LDL oxidation. Native LDL (n-LDL) was isolated from subjects on three long-term diets with differing fatty acid content (control diet rich in saturated fat and vegetarian and fish diets). The Cu2+-catalyzed oxidation of n-LDL was monitored using conjugated diene formation and stopped at various time points to achieve 20%, 45%, 70%, and 100% levels of ox-LDL. The lag phase of oxidation by Cu2+ was shortest and thiobarbituric acid-reactive substance (TBARS) formation by VSMC mediated oxidation was highest with n-LDL obtained from the fish diet group. There were no differences between the ox-LDLs obtained from the different diet groups in their cytotoxicity in EC culture. The degree of oxidation did not influence LDL cytotoxicity. In VSMC culture PGI2 production was increased by ox LDLs from all diet groups. In EC culture only the extensively oxidized LDLs obtained from the vegetarian diet group were able to induce PGI2 production. The LDLs did not affect basal cGMP production in either EC or VSMC culture. PMID- 10670835 TI - Thromboxane A2 receptor-mediated tonic contraction is attributed to an activation of phosphatidylcholine-specific phospholipase C in rabbit aortic smooth muscles. AB - Thromboxane A2 (TXA2) analogue STA2 produced a tonic contraction in rabbit aortic smooth muscles. In the present study, we examined phosphatidylcholine (PC) hydrolysis as a signaling pathway for the tonic contraction in rabbit aortic smooth muscles. In the primary cultured cells labeled with [3H]choline, STA2 caused an accumulation of [3H]phosphorylcholine, a metabolite of PC by PC specific PLC, in a concentration-dependent manner. The accumulation of [3H]phosphorylcholine was inhibited by SQ29548, a TXA2 receptor antagonist. In the muscle strips, STA2-induced tonic contraction was potently inhibited by D609, an inhibitor of PC-specific phospholipase C in a concentration-dependent manner with the IC50 of about 10 microM. Norepinephrine-induced tonic contraction was also inhibited by D609 with a weaker potency. These results strongly suggest that stimulation of TXA2 receptor results in the activation of PC-specific phospholipase C to yield diacylglycerol that contributes to the tonic contraction. PMID- 10670836 TI - Grepafloxacin inhibits tumor necrosis factor-alpha-induced interleukin-8 expression in human airway epithelial cells. AB - We examined the effect of grepafloxacin (GPFX), a new fluoroquinolone antimicrobial agent, on interleukin-8 (IL-8) expression in tumor necrosis factor alpha (TNF-alpha)-stimulated human airway epithelial cells (AEC). GPFX inhibited IL-8 protein production as well as mRNA expression in a concentration-dependent manner (2.5 - 25 micro g/ml), but the inhibition of IL-8 expression by corresponding concentrations of GPFX to serum and airway lining fluids was not complete. We discuss the modulatory effect of GPFX on IL-8 production in the context of its efficacy on controlling chronic airway inflammatory diseases. PMID- 10670837 TI - NF-kappaB regulatory mechanisms in alveolar macrophages from patients with acute respiratory distress syndrome. AB - Activation of the nuclear regulatory factor NF-kappaB occurs in the lungs of patients with the acute respiratory distress syndrome (ARDS) and may contribute to the increased expression of immunoregulatory cytokines and other proinflammatory mediators in this setting. Because of the important role that NF kappaB activation appears to play in the development of acute lung injury, we examined cytoplasmic and nuclear NF-kapppaB counterregulatory mechanisms, involving IkappaB proteins, in alveolar macrophages obtained from 7 control patients without lung injury and 11 patients with established ARDS. Cytoplasmic levels of the NF-kappaB subunits p50, p65, and c-Rel were significantly decreased in alveolar macrophages from patients with ARDS, consistent with enhanced migration of liberated NF-kappaB dimers from the cytoplasm to the nucleus. Cytoplasmic and nuclear levels of IkappaBalpha were not significantly altered in alveolar macrophages from patients with established ARDS, compared with controls. In contrast, nuclear levels of Bcl-3 were significantly decreased in patients with ARDS compared with controls (P = 0.02). No IkappaBgamma, IkappaBbeta, or p105 proteins were detected in the cytoplasm of alveolar macrophages from control patients or patients with ARDS. The presence of activated NF-kappaB in alveolar macrophages from patients with established ARDS implies the presence of an ongoing stimulus for NF-kappaB activation. In this setting, appropriate counterregulatory mechanisms to normalize nuclear levels of NF-kappaB and to suppress NF-kappaB-mediated transcription, such as increased cytoplasmic and nuclear IkappaBalpha levels or decreased Bcl-3 levels, appeared to be induced. Nevertheless, even though counterregulatory mechanisms to NF-kappaB activation are activated in lung macrophages of patients with ARDS, NF-kappaB remains activated. These results suggest that fundamental abnormalities in transcriptional mechanisms involving NF-kappaB and important in the inflammatory response occur in the lungs of patients with ARDS. PMID- 10670838 TI - Altered calcium regulation and function of human neutrophils during multiple trauma. AB - Altered intracellular Ca2+ concentration is a pivotal regulatory mechanism of leukocyte function. Since polymorphonuclear neutrophils (PMN) are involved in traumatic organ dysfunction, we prospectively investigated Ca2+ regulation and function of circulating PMN multiple trauma patients (Group A: ISS < 27; Group B: ISS > or = 27). Circulating PMN were isolated during 12 days, followed by determination of formyl-methionyl-leucyl-phenylalanine (fMLP)-induced PMN superoxide production (PMN-SOP) by SOD-inhibitable ferricytochrome C reduction, and PMN cytosolic Ca2+ concentration ([Ca2+]i) by fluorescent fura2/AM (340/380 ratio). PMN-SOP was significantly higher in Group B (mean ISS: 39.9 +/- 2; n = 21) at day of admission than in controls and Group A (mean ISS: 18.2 +/- 1; n = 22) (P< 0.05). In Group B, the significant rise of basal [Ca2+]i between Day 2 and Day 4 was associated with significant lower PMN-SOP during that period (P < 0.05). The fMLP-induced [Ca2+]i response was supranormal in both groups. PMN elastase concentrations were substantially higher in Group B compared with Group A until Day 4. Circulating IL-6, IL-8, and soluble TNF-receptor (55 kD) were significantly increased in Group B compared with Group A at the day of trauma (P < 0.05). Severe trauma is characterized by a biphasic pattern of neutrophil priming characterized by early increase and secondary suppression. The association of depressed neutrophil superoxide production (deactivation) and elevated basal [Ca2+]i suggests Ca2+-mediated disturbance of neutrophil NADPH oxidase metabolism. PMID- 10670839 TI - Peritonitis in the baboon: a primate model which stimulates human sepsis. AB - The physiological, hemostatic, and immunological responses of 12 chronically instrumented conscious baboons with sepsis due to Escherichia coli peritonitis were compared with that of similarly instrumented controls. Chronic indwelling cannulae were placed in the aorta and pulmonary artery to monitor pressure, cardiac output, and obtain blood samples. At t = 0 a sterile or E. coli-laden fibrin clot containing 1.9-6.7 x 10(11) CFU/kg was introduced into the peritoneal cavity. The control animals were group 1 (n = 3). The animals with peritonitis were divided into three groups depending on their clinical response. Group 2 animals (n = 3) were clinically well at the time of sacrifice (day 14), group 3 (n = 4) survived but were obviously sick on day 14, and group 4 (n = 5) died of sepsis. Implantation of a sterile fibrin clot was well tolerated with little hemodynamic change and a transient minimal inflammatory response in group 1. Implantation of an E. coli-containing clot elicited a hyperdynamic cardiovascular response and evoked a marked inflammatory reaction and a disseminated intravascular coagulopathy. Five of 12 (42%) E. coli animals died from sepsis. In general, the physiological, hemostatic, and immunological disturbances tended to be greatest in these animals. Autopsy revealed residual peritoneal inflammation and varying degrees of inflammation in the lungs, adrenal, spleen, liver, and kidneys in all the animals that received E. coli with the inflammatory infiltrate increasing in severity from group 2 through group 4. Tissue necrosis was observed only in the latter group. We conclude that the cardiovascular, hemostatic, and immunological responses of baboons with sepsis due to E. coli peritonitis exhibit a variable course that resembles the clinical manifestations of gram-negative sepsis in humans. PMID- 10670840 TI - Comparison of the mortality and inflammatory response of two models of sepsis: lipopolysaccharide vs. cecal ligation and puncture. AB - Sepsis remains a serious clinical problem despite intense efforts to improve survival. Experimental animal models of sepsis have responded dramatically to immunotherapy blocking the activity of cytokines. Despite these preclinical successes, human clinical trials have not demonstrated any improvement in survival. We directly compared the mortality, morbidity, and immunopathology in two models of sepsis, one due to lipopolysaccharide (LPS) and the other to cecal ligation and puncture (CLP). BALB/c mice were injected intraperitoneally with 250 microg of LPS or subjected to CLP with an 18-gauge needle. Both models yielded similar mortality (> 85%) and morbidity. Additionally, neutropenia and lymphopenia developed in both groups. Plasma and peritoneal levels of cytokines (TNF, IL-1, IL-6, and the chemokines KC and MIP-2) were measured at 1.5, 4, and 8 h after challenge. LPS induced substantially higher levels of cytokines in both compartments with peak levels between 1.5 and 4 h that began to decline at 8 h. In contrast, cytokine levels in the CLP model were continuing to increase at the 8 h-time point and often exceeded the LPS-induced values at this time. Our data demonstrate that the LPS and CLP models have similar mortality but significant differences in the kinetics and magnitude of cytokine production. Immunotherapy for sepsis based on cytokine production after LPS challenge is misdirected because the LPS model does not accurately reproduce the cytokine profile of sepsis. PMID- 10670841 TI - Pulmonary endothelial and epithelial integrity and neutrophil infiltration after endotoxin in interleukin-1 receptor knockout mice. AB - Previously we found the structural integrity of the aortic endothelium was maintained after the administration of endotoxin in type 1 interleukin-1 (IL-1) receptor knockout mice. In this study, we investigated further the integrity of pulmonary vascular endothelium, airway epithelial, pulmonary microvasculature, and neutrophil infiltration into the microvasculature and respiratory air spaces. Adult male C57BL/129J wild-type mice and C57BL/129J knockout mice possessing a homozygous deletion of the type 1 IL-1 receptor received the following intraperitoneal injections; 1) Escherichia coli endotoxin (ENDT) (10 mg/kg), 2) ENDT (2 mg/kg given for 4 days), or (3) saline vehicle. Wild-type and knockout control animals receiving saline vehicle showed normal endothelial and epithelial ultrastructure with intact membranes. Pulmonary endothelial cell damage was found only in wild-type mice given a single 10 mg/kg endotoxin dose. Airway epithelial damage was found only in wild-type mice given a repetitive dose of endotoxin (2 mg/kg for 4 days). Neutrophil infiltration increased only in mice given a single dose of endotoxin (10 mg/kg) with the wild-type increasing by 32% and the knockouts by 6% compared with the saline control for that group respectively. Serum IL-6 and nitric oxide (indicators of septic shock severity and lethality) significantly increased only in the mice given 10 mg/kg of endotoxin. The maintenance of pulmonary endothelial and epithelial cell integrity and the decrease of neutrophil infiltration in the IL-1 knockout mice suggest that IL-1 contributes significantly to the severity of endotoxin-induced sepsis. PMID- 10670842 TI - Effects of nicaraven on nitric oxide-related pathways and in shock and inflammation. AB - Expression of the inducible isoform of nitric oxide (NO) synthase, and the formation of peroxynitrite from NO and superoxide are responsible for some of the pathophysiological alterations seen during reperfusion injury and in various inflammatory conditions. Some of the effects of peroxynitrite are related to DNA single-strand breakage, and activation of poly (ADP-ribose) synthetase. Here we investigated the effect of nicaraven (2(R,S)-1,2-bis(nicotinamido)propane), a known hydroxyl radical scavenger compound and neuroprotective agent, on several NO- and peroxynitrite related pathways in vitro, and in shock and inflammation in vivo. Nicaraven, at 10 microM-10 mM, failed to inhibit the peroxynitrite-induced oxidation of dihydrorhodamine 123, indicating that the agent does not act as a scavenger of peroxynitrite. In RAW murine macrophages stimulated with peroxynitrite, nicaraven caused a dose-dependent, slight inhibition of poly (ADP ribose) synthetase activation, possibly due to a direct inhibitory effect on the catalytic activity of poly (ADP-ribose) synthetase. Nicaraven partially protected against the peroxynitrite-induced suppression of mitochondrial respiration in RAW macrophages and caused a slight, dose-dependent inhibition of nitrite production in RAW macrophages stimulated with bacterial lipopolysaccharide. We next investigated the effect of nicaraven treatment in a variety of models of inflammation and reperfusion injury. Nicaraven (at 10-100 microg/paw) exerted significant protective effects in the carrageenan-induced paw edema model and (at 100 mg/kg i.v.) reduced neutrophil infiltration and histological damage in splanchnic artery occlusion-reperfusion injury. However, nicaraven failed to alter the course of hemorrhagic and endotoxic shock and arthritis in rodent models. The current data indicate the limited role of hydroxyl radicals in the pathogenesis of the inflammatory conditions tested. PMID- 10670843 TI - Nitric oxide synthase inhibitor ameliorates oral total parenteral nutrition induced barrier dysfunction. AB - The expression of inducible nitric oxide synthase (iNOS) is increased in the intestine and results in mucosal damage after endotoxin challenge. Although the oral administration of total parenteral nutrition (TPN) solution promotes bacterial translocation (BT) and increases the intestinal permeability, the role of NO in the nutrition-induced loss of mucosal barrier function remains unclear. The distribution of fluorescein isothiocyanate-dextran (FITC-dextran, 4400) across the lumen of small intestine in rat was examined to investigate the role of NOS activity on the intestinal permeability under oral TPN feeding. Fifty-one rats were randomly divided into 4 groups. Group I (control group) was fed with rat chow, group II received TPN solution orally. Groups III and IV received TPN solution supplemented with NOS inhibitors. On day 9, FITC-dextran was injected into the intestinal lumen. After 30 min, blood samples were taken from portal vein and analyzed for plasma FITC-dextran level by fluorescence spectrophotometry. Samples of small intestine were frozen and sectioned in a cryostat for morphological and NOS histochemical studies. Homogenates of small intestine were used for NOS activity measurement. The plasma level of FITC dextran showed a significant increase (P < 0.05) in rats fed with oral TPN compared with the control ones. Supplement with NOS inhibitors significantly decreased the intestinal permeability in groups III and IV compared with group II. Similarly, the total NOS activities showed a significant 2-fold increase (P< 0.05) in group II, and NOS inhibitors decreased the elevated NOS activity. These data suggest that oral TPN feeding for 9 days leads to an increase in permeability to dextran and the total NOS activity of small intestine, and both induction of the intestinal permeability and NOS activity were inhibited by treatment with NOS inhibitors. Addition of S-methylisothiourea (SMT), an iNOS selective inhibitor, profoundly inhibited 66% of the induced iNOS activity (P < 0.05) and reduced 74% of the diet-induced increase in intestinal permeability (P < 0.05) in group II. The induced permeability change in rats receiving oral TPN is mainly due to the activity of intestinal mucosal iNOS. The induction of iNOS is an important mediator for intestinal barrier dysfunction. Administration of SMT, which specifically decreases iNOS activity, is useful in the prevention of diet-induced barrier failure. PMID- 10670844 TI - Kinetics of P-selectin expression in regional vascular beds after resuscitation of hemorrhagic shock: a clue to the mechanism of multiple system organ failure. AB - Leukocyte-endothelial cell interactions play an important role in mediating organ dysfunctions observed after hemorrhagic shock. P-selectin is the first endothelial cell adhesion molecule to be upregulated after an ischemic insult. The objective of this study was to define kinetics of P-selectin expression in different regional vascular beds of mice exposed to hemorrhagic shock. In-vivo P selectin expressions were determined using dual radiolabeled monoclonal antibody technique in lungs, heart, liver, kidneys, intestinal mesentery, stomach, small bowel, and colon 0.5, 1, 2, 5, 10, and 24 h after resuscitation of 40 mmHg hemorrhagic shock. In another group, P-selectin expression was determined in same organs 5 h after resuscitation of 30 mmHg hemorrhagic shock. Hemorrhagic shock of 40 mmHg caused significant upregulation of P-selectin in lungs and liver at 30 min after resuscitation (P < 0.001). There was a second and more pronounced upregulation of P-selectin in lungs and liver at 5 h after resuscitation (P < 0.001). In heart, intestinal mesentery, stomach, small bowel, and colon, P selectin was not upregulated until 5 h after resuscitation from 40 mmHg hemorrhagic shock (P < 0.001). While hemorrhagic shock of 40 mmHg did not cause P selectin upregulation in kidneys, hemorrhage to 30 mmHg did elicit a significant increase at 5 h after resuscitation (P < 0.001). We conclude that P-selectin is upregulated after resuscitation of hemorrhagic shock in lungs, liver, heart, stomach, and intestines. P-selectin upregulation in kidneys only takes place after more severe hemorrhagic shock. PMID- 10670845 TI - A role for nitric oxide in endotoxin-induced depletion of the peripheral catecholamine stores. AB - Endotoxemia is associated with increased sympathetic nerve activity and depletion of norepinephrine (NE) and epinephrine (EPI) contents in the adrenal gland and in sympathetically innervated tissues. Endotoxin (bacterial lipopolysacchride [LPS]) causes an increased production of nitric oxide (NO) by inducing nitric oxide synthase (iNOS) expression in various tissues. This increased production of NO contributes significantly to the hypotension associated with endotoxemia. At high concentrations, NO also can act as a neurotoxin. In this study we tested the hypothesis that increased production of NO is involved in depletion of catecholamine content in various tissues from rats treated with a nonlethal dose of LPS. Catecholamine content was measured by high-performance liquid chromatography with electrochemical detection (HPLC-EC) and NOS activity was measured by the 3H-I-arginine to 3H-I-citrulline conversion method. The NE content was decreased in rat adrenal gland, lung, spleen, tail artery, and aorta after LPS. The maximal decrease was at 24 h and returned to control levels at 6 days (144 h). There was no depletion of the NE content in the heart. The EPI content in the adrenal gland was greatly depleted (91%) from 12 to 72 h after LPS. LPS increased the NOS activity in all tissues examined. The time course for NOS activity showed an increase at 3 h, a further increase at 6 h, and a return to control level at 48 h after LPS. The increase in NOS activity occurred before maximal catecholamine depletion. Aminoguanidine, a relatively selective iNOS inhibitor, completely prevented NE depletion in all tissues and partially prevented adrenal EPI depletion induced by LPS. These results are consistent with the hypothesis that LPS-induced production of NO plays a role in depletion of tissue NE and EPI. PMID- 10670846 TI - Bovine hemoglobin-based oxygen carrier (HBOC-201) for resuscitation of uncontrolled, exsanguinating liver injury in swine. Carolina Resuscitation Research Group. AB - In the setting of rapidly exsanguinating hemorrhage, resuscitation with intravenous (i.v.) crystalloid solution may not sustain survival before availability of allogenic blood transfusion and surgery. This study tested the hypothesis that bovine hemoglobin-based oxygen carrier, HBOC-201, would improve resuscitation and extend early survival from exsanguinating hemorrhage. This study simulated the prehospital scenario of rapidly exsanguinating hemorrhage with prolonged prehospital time and lack of blood availability. Severe hemorrhagic shock was induced in swine by using multiple liver lacerations. At 9 min after the onset of bleeding, swine were randomized to receive approximately 10 mL/kg/min of i.v. lactated Ringer's solution (n = 10) or HBOC-201 (n = 7) to achieve a mean aortic pressure (MAP) of 60 mmHg. Thereafter, infusion rate was adjusted to maintain MAP at 60 mmHg for up to 2 h. All animals were initially successfully resuscitated. The results showed 2-h survival was 1 of 10 with lactated Ringer's and 7 of 7 with HBOC-201 (P = 0.0004). Nine lactated Ringer's swine had cardiovascular collapse at 36 +/- 10 min. Lactate at 30 min was 18 +/- 3 mmol/L with lactated Ringer's and 12 +/- 2 mmol/L with HBOC-201 (P < 0.05). Hematocrit was <1% in 9 of 10 lactated Ringer's and 6 of 7 HBOC-201 animals. These data indicate that HBOC-201 improved early survival and stabilized hemodynamic and metabolic parameters vs. lactated Ringer's in this swine model of liver injury with uncontrolled, lethal hemorrhage that simulates the prehospital care environment where allogenic blood is unavailable. PMID- 10670847 TI - Suppression of lethal toxicity of endotoxin by biscoclaurine alkaloid cepharanthin. AB - Suppressive effects of Cepharanthin (CE) on lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNFalpha) production followed by liver injury were investigated. Pretreatment with CE reduced limulus activity of LPS. Intraperitoneal treatment with CE 10 min before an i.v. challenge of LPS resulted in protection from LPS lethality in D-galactosamine (GalN)-sensitized BALB/c but not in C57BL/6 and C57BL/10ScSn mice. Treatment with CE before the LPS challenge significantly reduced serum TNF levels in a dose-dependent manner. The suppression was most effective when CE was administered 10 min before the LPS challenge. Increased levels of enzymes released from hepatocytes into the circulation, as a result of LPS-induced liver injury, were reduced by CE administration. Histological evaluation demonstrated that massive cell infiltration after severe injury developed in liver of mice injected with LPS plus D-GalN unless they were pretreated with CE. Apoptotic cells decreased by treatment with CE. Treatment with CE retarded lethal shock induced by an infection with 10(8) CFU Salmonella typhimurium deltaaroA mutant. These results suggest that action of CE is initiated through suppression of LPS-induced TNF production. PMID- 10670848 TI - Strategic planning for survival of surgical societies in the new millennium. PMID- 10670849 TI - Surgical progress and understanding in the treatment of the melanoma epidemic. PMID- 10670850 TI - The role of the gastrointestinal tract in postinjury multiple organ failure. AB - Despite intensive investigation, the pathogenesis of postinjury multiple organ failure (MOF) remains elusive. Laboratory and clinical research strongly implicate that the gastrointestinal tract plays a pivotal role. Shock with resulting gut hypoperfusion appears to be one important inciting event. While early studies persuasively focused attention on bacterial translocation as a unifying mechanism to explain early and late sepsis syndromes that characterize postinjury MOF, subsequent studies suggest that other gut-specific mechanisms are operational. Based on our Trauma Research Center observations and those of others, we conclude that: 1) bacterial translocation may contribute to early refractory shock; 2) for patients who survive shock, the reperfused gut appears to be a source of proinflammatory mediators that may amplify the early systemic inflammatory response syndrome; and 3) early gut hypoperfusion sets the stage for progressive gut dysfunction such that the gut becomes a reservoir for pathogens and toxins that contribute to late MOF. PMID- 10670851 TI - Radiation safety with breast sentinel node biopsy. AB - BACKGROUND: Sentinel lymph node biopsy with Technetium 99m sulfur colloid (Tc99m) is an evolving technique that offers the potential for improved staging of breast cancer with decreased morbidity. However, the use of radioactive materials in the operating room generates significant concern about radiation exposure. The purpose of this study was to evaluate radiation exposure to operating room personnel, pathologist, and equipment from specimens during breast sentinel lymph node biopsy. METHODS: Twenty patients were injected with 0.7 to 1.1 mCi of Tc99m sulfur colloid 1.5 to 3 hours before sentinel lymph node biopsy. A calibrated Geiger counter was used to measure dose rates from the breast injection site before skin incision (n = 20), lumpectomy specimens (n = 8), and sentinel nodes (n = 20) at distances of 3, 30, and 300 cm. This represented exposure to the surgeon's hands, surgeon's torso, and scrub nurse, respectively. Exposure to the pathologist's hands and torso was represented as dose-rate measurements from lumpectomy and nodal specimens. The operative instruments, trash receptacles, suction canisters, pathology slides, and cryostat machines were measured at 3 cm at the conclusion of each procedure. Specimens or equipment emitting radiation doses equal to background levels (0.04 mRem/h) were exempt from special handling and disposal. RESULTS: The highest exposure rate was to the surgeon's hands from the breast injection site before skin incision (34.25 mRem/h). Exposure to the surgeon's torso measured 1.33 mRem/h, and exposure to the scrub nurse's torso measured 0.15 mRem/h from the injection site. Exposure to the pathologist's hands was 18.62 and 0.06 mRem/h from the lumpectomy specimen and sentinel node, respectively. Exposure to the pathologist's torso measured 0.34 and 0.04 mRem/h from the lumpectomy specimen and sentinel node, respectively. One hundred percent of lumpectomy specimens measured above the exempt level. Thirty-two of 46 (70%) sentinel lymph nodes emitted radiation equal to the exempt background level. Seventeen of 20 trash receptacles (85%) and 4 of 12 (33%) suction canisters measured equal to background levels. All operative instruments, pathology slides, and cryostat machines were equal to background levels. CONCLUSIONS: Radiation exposure to operating room personnel, pathologists, and operative equipment during a breast sentinel node biopsy using Tc99m is minimal. A primary surgeon can perform 2,190 hours, a scrub nurse 33,333 hours, and a pathologist 14,705 hours of procedural work before surpassing Occupational Safety and Health Administration limits. Operative instruments, pathology slides, and cryostat machines do not require special handling. All lumpectomy specimens should be stored for decontamination until the dose rate equals background levels. Intraoperative dose-rate monitoring allows selective decontamination of nodal specimens, trash receptacles, and suction canisters, which decreases disposal time and cost. PMID- 10670852 TI - Is laparoscopic approach to lumbar spine fusion worthwhile? AB - BACKGROUND: Laparoscopic lumbar spine fusion has been recently described. The aim of this study is to evaluate the safety and efficacy of this procedure for single and multiple-level degenerative disc disease. METHODS: Twenty-four consecutive laparoscopic interbody lumbar fusions were evaluated prospectively (18 single level were compared with 6 multiple-level procedures). Results of the laparoscopic multiple-level procedures were further compared with 12 open multiple-level operations. RESULTS: Twenty procedures were completed laparoscopically. The conversions were related to iliac vein lacerations (3 cases) and a mesenteric tear. Single-level cases had lower morbidity (22% versus 83%), shorter hospital stay (2 versus 10 days), and higher fusion rate (88% versus 50%) than multiple-level procedures. Overall results in the latter group were worse than in the matched open group. CONCLUSIONS: Laparoscopic single-level fusion (L5-S1) is safe and carries the benefits of minimal access surgery. Morbidity after multiple level approach is high, and this procedure cannot be advocated at this time. PMID- 10670853 TI - Cholecystectomy is an effective treatment for biliary dyskinesia. AB - BACKGROUND: An increasing number of reports indicate symptomatic relief of biliary colic symptoms after cholecystectomy for biliary dyskinesia. Despite this, cholecystectomy as a treatment for biliary dyskinesia remains controversial. Our aim was to determine efficacy of cholecystectomy in alleviating biliary dyskinesia symptoms and the correlation with histologic findings. METHODS: Records of patients with gallbladder ejection fraction <35% between January 1994 and February 1999 were reviewed. Gallbladder pathology and degree of symptomatic improvement were determined on follow-up. RESULTS: Of the 27 cholecystectomy patients, 24 (89%) had significant improvement, 2 (7%) had partial improvement, and 1 (4%) had minimal improvement. Ten patients (43%) had normal gall-bladder, and 9 (90%) of them had significant improvement after cholecystectomy. Of the 6 nonsurgical patients, none had significant improvement, 4 (67%) had partial improvement, and 2 (33%) had minimal improvement. CONCLUSIONS: Biliary dyskinesia patients who underwent cholecystectomy had significantly greater symptom improvement compared with nonsurgical patients. Pathologic correlation suggests chronic inflammation may not be the only cause of gallbladder dysfunction. Cholecystectomy should be a first-line therapy for biliary dyskinesia patients. PMID- 10670854 TI - Breast conservation therapy in affiliated county, university, and private hospitals. AB - BACKGROUND: Breast conservation therapy (BCT) offers equivalent survival to modified radical mastectomy in patients with early-stage (I and IIa) breast cancer, but is utilized in less than 50% of eligible patients. While patient demographics have been linked to BCT rates, we suspected that physician influence was a major factor. The purpose of this study was to compare BCT at three affiliated centers staffed by similarly trained surgeons yet serving widely disparate populations, in order to assess the importance of physician influence on the utilization of BCT. METHODS: Tumor registry data were reviewed from 1993 through 1997 at affiliated city/county (CH), university (UH), and private hospitals (PH). Data were analyzed for clinical stage, treatment, and age of patient. RESULTS: The utilization of BCT for stage I and IIa breast cancer is similar at the three hospitals: 45% of patients at CH, 55% of patient at UH, and 57% of patients at PH (P>0.05). Rates of BCT were similar across all patient age groups at all sites. CONCLUSIONS: Similar BCT utilization rates can be achieved despite widely disparate patient populations. The three affiliated hospitals are staffed by surgeons with similar training, and all offer a multidisciplinary approach to breast cancer care. This suggests that physician influence may override patients' socioeconomic issues in providing optimal breast cancer therapy. PMID- 10670855 TI - Low use of breast conservation surgery in medically indigent populations. AB - BACKGROUND: Breast conservation surgery (BCS) with radiation is an acceptable treatment for early-stage breast cancer. METHODS: Data were obtained from hospital cancer registries on women surgically treated for Stage 0 to II breast cancer from 1993 to 1997. Data on 1,747 patients were analyzed for surgical treatment, hospital type (private versus public), disease stage, and ethnic origin. RESULTS: In this study, 34% of women received BCS. Women treated in private hospitals received BCS more often than women treated in public hospitals. Women with stage II disease received BCS less often than women with earlier stage disease. Hospital type (public versus private) and disease stage were strong, independent predictors for use of BCS. When hospital type and disease stage were statistically controlled, no treatment differences across ethnic groups were identified. CONCLUSIONS: Use of BCS in this study was low compared with National Cancer Database statistics. Women treated in publicly funded hospitals and those with stage II disease were significantly less likely to receive BCS. PMID- 10670856 TI - Accuracy of surgeon-performed gallbladder ultrasound. AB - BACKGROUND: Symptomatic cholelithiasis is among the most common of general surgery referrals. With an appropriate clinical presentation, definitive diagnosis requires documentation of gallstones by ultrasonography (US). The authors evaluated the accuracy of surgeon-performed US for identifying gallstones in patients with a nonacute indication for study. METHODS: Patients referred for symptomatic cholelithiasis and who provided informed consent received an US examination by one or more of the surgical investigators. Surgeon-performed US findings were correlated with radiologist US findings and pathologic diagnoses. RESULTS: Seventy-seven patients received a total of 128 examinations by the investigators. Surgeon-performed US examination agreed with the radiologist US findings for 112 of 122 studies (92%) with a sensitivity of 100% and a specificity of 95%. Surgeon-performed US findings correlated with the pathologic diagnoses for 83 of 86 studies (97%). CONCLUSIONS: Surgeons can perform gallbladder US in the nonacute setting with a high degree of accuracy. PMID- 10670857 TI - The efficacy of magnetic resonance cholangiography for the evaluation of patients with suspected choledocholithiasis before laparoscopic cholecystectomy. AB - BACKGROUND: Endoscopic retrograde cholangiography is the most commonly utilized tool for the identification of common bile duct stones (CBDS) before laparoscopic cholecystectomy, whereas the role of magnetic resonance cholangiography (MRC) for patient evaluation before laparoscopic cholecystectomy is currently undefined. METHODS: We prospectively evaluated the efficacy of MRC for the identification of CBDS among patients with high risk for choledocholithiasis. Patient selection was based on clinical, sonographic, and laboratory criteria. Standard cholangiograms were obtained when possible for verification of MRC results. RESULTS: Ninety-nine patients underwent evaluation with preoperative MRC. CBDS was visualized in 30% of patients. MRC sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 85%, 90%, 77%, 94%, and 89%, respectively. CONCLUSIONS: MRC is useful for the evaluation of patients with suspected choledocholithiasis. Advantages of MRC include its noninvasive nature, ease of application, and accuracy in identifying and estimating the size of CBDS. Application of MRC in this setting reduces the need for diagnostic endoscopic retrograde cholangiography. Future investigations should be directed at the development of cost-effective utilization strategies for MRC application. PMID- 10670858 TI - The role of computed tomography in the diagnosis of acute appendicitis. AB - BACKGROUND: Routine contrast-enhanced computed tomography (CECT) has been described as an accurate diagnostic imaging modality in patients with acute appendicitis. However, most patients with acute appendicitis can be diagnosed by clinical findings and physical exam alone. The role of CECT in patients suspected of having appendicitis but with equivocal clinical exams remains ill defined. METHODS: One hundred and seven consecutive patients who were thought to have appendicitis but with equivocal clinical findings and/or physical exams were imaged by CECT over a 12-month period. Oral and intravenous contrast-enhanced, spiral abdominal and pelvic images were obtained using 7-mm cuts. CECT images were interpreted by a board-certified radiologist. Main outcome measures included CECT sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy in the diagnosis of acute appendicitis, comparing CECT with ultrasound, and determining the impact of CECT on the clinical management of this patient population. RESULTS: A group of 107 patients consisting of 44 males (41%) and 63 females (59%) with a median age of 33 years (range 13 to 89 years) were imaged with CECT to evaluate suspected appendicitis. Of the 107 CECTs performed, 11 false-positive and 3 false-negative readings were identified, resulting in a sensitivity of 92%, specificity of 85%, PPV of 75%, NPV of 95%, and an overall accuracy of 90%. Forty-three patients were imaged with ultrasound and CECT, and CECT had significantly better sensitivity and accuracy (30% versus 92% and 69% versus 88%, P<0.01). With regard to clinical management, 100% (36/36) of patients with appendicitis, and 4.2% (3/71) of patients without appendicitis underwent appendectomy. Therefore, the overall negative appendectomy rate was 7.6% (3/39). CONCLUSIONS: CECT is a useful diagnostic imaging modality for patients suspected of having acute appendicitis but with equivocal clinical findings and/or physical exams. CECT is more sensitive and accurate than ultrasound and is particularly useful in excluding the diagnosis of appendicitis in those without disease. PMID- 10670859 TI - Laparoscopic ultrasound imaging of adrenal tumors during laparoscopic adrenalectomy. AB - BACKGROUND: The purpose of this study was to determine the usefulness of laparoscopic ultrasound (LUS) during laparoscopic adrenalectomy (LA) and to define the ultrasound imaging characteristics of various adrenal tumors. METHODS: LUS was utilized in 27 patients who underwent LA (including one bilateral adrenalectomy) from May 1994 to October 1998. Tumor size ranged from 1.0 to 5.5 cm (mean 3.3 cm), and a transabdominal lateral approach to LA was used. RESULTS: LUS localized the adrenal gland and tumor in all 28 adrenalectomies and demonstrated the relationship of the tumor to the kidney and adjacent vascular structures (renal artery/vein and inferior vena cava). The adrenal vein was visualized sonographically in only six cases (21 %). Pheochromocytomas were mild to markedly heterogenous, whereas most aldosteronomas and cortical adenomas were homogenous. LUS provided useful information to the surgeon in 11 of 28 cases (39%) by: 1) localizing the adrenal gland and tumor and/or guiding the dissection; 2) demonstrating that tumors > or =4 cm were confined to the adrenal gland; and 3) investigating suspected pathology in other organs. Mean operating time for LUS was 10.9 min (range 5 to 24 min) and calculated hospital charges were $602. CONCLUSIONS: LUS accurately localizes adrenal tumors, helps define their relationship to adjacent structures, and provides confirmation that larger tumors are amenable to laparoscopic resection. LUS is a useful adjunct to laparoscopic adrenalectomy in selected patients. PMID- 10670860 TI - Role of breast magnetic resonance imaging in determining breast as a source of unknown metastatic lymphadenopathy. AB - BACKGROUND: Occult primary breast cancer (OPBC) represents less than 1% of breast cancer. In only a third of cases, mammography identifies a primary tumor. We hypothesized that rotating delivery of excitation off-resonance breast magnetic resonance imaging (MRI) would identify or exclude the breast as a primary site in patients with OPBC. METHODS: In a retrospective review, 10 patients were identified with OPBC in which MRI was performed. Malignant appearing lesions were correlated with histopathologic findings at biopsy or surgery. RESULTS: MRI identified the primary site in 8 of 10 cases as breast (80%), and excluded it in 2 cases. The extent of disease and location was accurately predicted when compared with histopathologic specimen. CONCLUSIONS: As we continue to focus on a cure of early breast cancer, it is imperative that diagnostic images become more sensitive and specific. MRI accurately predicted OPBC in this subset of patients. PMID- 10670861 TI - Complete myocardial revascularization on the beating heart. AB - BACKGROUND: Complete myocardial revascularization with excellent visualization, exposure, and stabilization can be accomplished on the beating heart without cardiopulmonary bypass (CPB). METHODS: Three hundred patients were totally revascularized via median sternotomy with myocardial stabilization using the CardioThoracic System. All patients who underwent coronary artery bypass grafting were considered for the off-pump procedure. Pericardial sutures were placed at the level of the left atrial appendage and were pulled upwards to the right. The stabilizer was applied sequentially from circumflex, obtuse marginal, intermediate, diagonal, left anterior descending, and right coronary artery. Coronaries were occluded using the Calafiore technique, and multiple arterial grafts were inserted. RESULTS: The average number of grafts was 3.4 per patient. Six percent had to be converted to standard CPB. Comorbidity was not a limiting factor with 8% redos, 48% having diabetes, and acute myocardial infarctions in 28%. The unadjusted mortality was 2.3%, and stroke rate was 0.7%. CONCLUSIONS: These results indicate that complete revascularization can safely be accomplished without CPB. PMID- 10670862 TI - Comparison of carotid endarterectomy using primary closure, patch closure, and eversion techniques. AB - BACKGROUND: This study was undertaken to evaluate the role of eversion endarterectomy in the management of extracranial carotid occlusive disease. METHODS: A retrospective review was performed of all patients undergoing carotid endarterectomy between July 1994 and July 1998. After reviewing the records, patients were assigned to one of three groups: eversion (ECEA); open with primary closure (CEA); or open with patch closure (CEAP). Statistical comparisons were made. RESULTS: The 190 index cases comprised 33 ECEA (17%), 15 CEA (8%), and 142 CEAP (75%). Both ECEA and CEA were more likely to be done on males versus females compared with CEAP (P = 0.01). For the entire 190 cases, stroke occurred in 1 patient (0.5%); and myocardial infarction in 2 patients (1%), resulting in death in both. Two patients (1.4%) in the CEAP group have undergone redo surgery at 8 and 24 months. CONCLUSIONS: This study demonstrates that eversion endarterectomy achieves early results similar to open endarterectomy with and without patch closure. PMID- 10670863 TI - Surgical complications from hemostatic puncture closure devices. AB - BACKGROUND: For securing immediate hemostasis following percutaneous arterial catheterization, the Food and Drug Administration has approved three hemostatic puncture closure devices. We reviewed our institutional experience with one device (Angio-Seal). METHODS: A retrospective, single-center, nonrandomized observational study was made of all vascular complications following femoral cardiac catheterization. RESULTS: An immediate mechanical failure of the device was experienced in 34 (8%) patients. Surgical repair was required in 1.6% (7 of 425) of patients following Angio-Seal versus 0.3% (5 of 1662) following routine manual compression (P = 0.004). In 5 patients, the device caused either complete occlusion or stenosis of the femoral artery. The polymer anchor embolized in 1 patient and was retrieved with a balloon catheter at surgery. CONCLUSION: During the first year of utilization of a percutaneous hemostatic closure device following cardiac catheterization, we observed a marked increase in arterial occlusive complications requiring surgical repair. Surgeons must be familiar with the design of these devices to achieve precise repair of surgical complications. PMID- 10670864 TI - Optimizing screening for blunt cerebrovascular injuries. AB - BACKGROUND: The recognition that early diagnosis and intervention, prior to ischemic neurologic injury, has the potential to improve outcome following blunt cerebrovascular injuries (BCVI), led to a policy of aggressive screening for these injuries. The resultant epidemic of BCVI has created a dilemma, as widespread screening is impractical. We sought to identify independent predictors of BCVI, to focus resources. METHODS: Cerebral arteriography was performed based on signs or symptoms of BCVI, or in asymptomatic patients with high-risk mechanisms (hyperextension, hyperflexion, direct blow) or injury patterns. Logistic regression analysis identified independent predictors. RESULTS: A total of 249 patients underwent arteriography; 85 (34%) had injuries. Independent predictors of carotid arterial injury were Glasgow coma score < or =6, petrous bone fracture, diffuse axonal brain injury, and LeFort II or III fracture. Having one of these factors in the setting of a high-risk mechanism was associated with 41% risk of injury. Of patients with cervical spine fracture, 39% had vertebral arterial injury. CONCLUSIONS: Patients sustaining high-risk injury mechanisms or patterns should be screened for BCVI. In the face of limited resources, screening efforts should be focused on those with high-risk predictors. PMID- 10670865 TI - The laparoscopic management of appendicitis and cholelithiasis during pregnancy. AB - BACKGROUND: Laparoscopic management of appendicitis and symptomatic cholelithiasis during pregnancy remains controversial. We report the single largest series of laparoscopic cholecystectomies and appendectomies during pregnancy. METHODS: Medical records of all pregnant patients who underwent open or laparoscopic management of appendicitis/cholelithiasis at LDS Hospital from 1990 to 1998 were reviewed. RESULTS: Eighteen open appendectomies (OA) and 13 open cholecystectomies (OC) were performed. Forty-five laparoscopic cholecystectomies (LC) and 22 laparoscopic appendectomies (LA) were performed without birth defects, fetal loss or uterine injury. Preterm delivery rates (PTD) in the LA and OA groups were similar (15.8% versus 11.8%, P>0.9). The PTD rate in the LC group was not significantly different than in the OC group (11.9% versus 10.0%, P>0.9). Neither birth weights nor Apgar scores were significantly different across groups. CONCLUSIONS: Laparoscopic management of appendicitis and symptomatic cholelithiasis during pregnancy can be performed with minimal fetal and maternal morbidity when accepted management guidelines are followed. PMID- 10670866 TI - Laparoscopic splenectomy in patients with hematologic malignancies. AB - BACKGROUND: Although laparoscopic splenectomy (LS) for benign hematologic disease is well accepted, its role in hematologic malignancies is not clearly defined. This study examined the efficacy and feasibility of LS for hematologic malignancies. METHODS: Records were reviewed from patients who underwent LS at two university hospitals. Charts from 77 open splenectomies for malignancy (OM) during the same period were also reviewed. RESULTS: Fifty-three patients underwent LS, 22 for hematologic malignancies (LM) and 31 for benign hematologic disorders (LB). Median splenic weight was greater in the LM group (930 g) than in the LB group (164 g, P = 0.001). LM was associated with longer operations and greater blood loss than was LB. LM had a 41% conversion rate. Morbidity, mortality, and transfusion rates were similar. Median hospital stay was shorter for LM (4 days) than for OM (6 days, P = 0.001). CONCLUSIONS: LS is feasible in hematologic malignancies but is associated with increased operative time and blood loss and a high conversion rate. Morbidity and mortality, however, was similar. Shorter hospital stays for LM compared with OM may translate into earlier recovery and initiation of antineoplastic therapy. PMID- 10670867 TI - A state-wide evaluation of appendectomy in children. AB - BACKGROUND: Traditional management of appendicitis in children involves open appendectomy (OA), an operation that is relatively inexpensive and carries few risks and complications. However, little information is available regarding the use, cost, and complication of laparoscopic appendectomy (LA) in children. METHODS: Our initial aim was to determine if LA is frequently performed in children (<15 years). We then compared the surgical results of OA versus LA. In conjunction with the Missouri Department of Health, we evaluated 793 children treated for appendicitis throughout the state between January 1997 and June 1997. The authors were blinded to the patient, surgeon, and hospital; no children were excluded. RESULTS: LA was infrequently performed in children with advanced disease. Overall, children undergoing LA were older and had a shorter hospitalization but no difference in hospital charge. When separated by child age, LA was associated with a shorter length of stay in all groups (0 to 5, 6 to 10, and 11 to 15 years) but only children in the 6 to 10 year range had a lower hospital charge when compared with patients undergoing OA. CONCLUSIONS: LA is becoming a common surgical approach for older children with simple appendicitis. Furthermore, these data suggest that LA, independent of individual surgeon or medical center, is associated with a decreased length of hospitalization without a significant difference in hospital charge. PMID- 10670868 TI - Laparoscopic refundoplications after failed antireflux surgery. AB - BACKGROUND: Open and laparoscopic antireflux procedures may require reoperation for failures of the initial procedure in about 3% to 6% of cases. The purpose of this study is to describe our operative experiences, postoperative results, and patients' view of outcome following laparoscopic refundoplication. METHODS: Thirty patients (18 men, 12 women), mean age 56 years (range 37 to 77) underwent laparoscopic redo surgery. In 18 patients the initial surgery was done by the open technique, and 3 had surgery twice previously. Twelve patients had previous laparoscopic antireflux surgery. Indications for redo surgery were recurrent reflux (n = 17), dysphagia (n = 6), and the combination of both (n = 7). RESULTS: Twenty-eight patients were completed laparoscopically, 21 with a floppy Nissen and 7 with a Toupet fundoplication. Two patients were converted to the open procedure because of intraoperative technical problems. In 5 cases there was an injury to the stomach wall, successfully managed laparoscopically. Postoperatively 1 patient had dysphagia and required pneumatic dilatation, another had gas bloat. There was a significant increase in lower esophageal sphincter pressure at 3 months (12.4+/-4.8 mm Hg; n = 30) and 1 year (12.3+/-4.5 mm Hg; n = 30). Twenty-four hour pH monitoring showed a decrease of the DeMeester Score at 3 months after surgery from 14.7+/-10.6 (n = 30) and 1 year after surgery from 12.1+/-8.7 (n = 30). Gastrointestinal quality of life index increased from 87 points preoperatively to 121 at 3 months and 123 at 1 year, which is comparable with a healthy population (123 points). CONCLUSIONS: Laparoscopic refundoplication is a feasible and effective procedure with excellent postoperative results, independent of whether the primary procedure was done by the open or laparoscopic technique. PMID- 10670869 TI - Surgical management of biliary gallstone disease during pregnancy. AB - BACKGROUND: Biliopancreatic gallstone disorders (BPD) manifesting during pregnancy are relatively rare. The management of these conditions remains controversial. Although perioperative problems and fetal loss have been reported, recent publications have advocated an early surgical approach. PATIENTS AND METHODS: Thirty-two pregnant women underwent operation for BPD between January 1993 and December 1997. The mean age was 29 years and ranged from 18 to 41 years. RESULTS: Twelve patients underwent a laparoscopic cholecystectomy (LC), and 20 open cholecystectomies (OC), including two conversions from laparoscopic. Seven of the OC patients required additional open CBD exploration and intraoperative choledochoscopy for CBD stones. No maternal mortality was observed. A single fetal demise (3%) occurred for a patient with gallstone pancreatitis who underwent open cholecystectomy during her 14th week of gestation. CONCLUSIONS: Early involvement of the obstetric team, with preoperative and postoperative fetal monitoring, and adequate management of anesthetic and tocolytic agents make cholecystectomy a safe procedure at any stage of pregnancy. PMID- 10670870 TI - Autologous blood transfusion does not reduce postoperative infection rates in elective surgery. AB - BACKGROUND: The influence of blood transfusions in the risk of postoperative infection remains controversial. We examined the association between autologous (AB) and homologous (HB) blood transfusions with postoperative infection in elective surgery. METHODS: The medical records of 991 Medicare patients aged > or =65 years submitted to hysterectomy and hip and knee replacement were reviewed. Logistic regression analysis was used to control for age, comorbidity, year, and type of procedure. RESULTS: Overall, 451 (46%) patients required transfusions. AB was given to 324 (72%), HB to 94 (21%); 33 (7%) patients received both. Forty-two patients (4%) developed postoperative infections. The infection rate was not different among patients receiving HB (7%), AB (5%), AB+HB (0), and nontransfused patients (4%); P = 0.18). After adjustment for confounders, HB and AB remained not associated with infections. CONCLUSION: In elective surgery with small volume transfusion, neither AB nor HB transfusions were associated with an increased risk of postoperative infections. PMID- 10670871 TI - Attenuating tumor necrosis factor alpha does not ameliorate other cytokine and peroxidase products during sepsis. AB - BACKGROUND: Recent trials utilizing single anticytokine agents have shown no consistent survival benefit in improving the outcome of sepsis. Since an entire cascade of mediators contributes to the underlying pathophysiology, it is not surprising that monotherapy has proven unsuccessful. The purpose of this study was to measure the effects of attenuating tumor necrosis factor (TNF)alpha early in sepsis. METHODS: Three groups of Sprague-Dawley rats were studied. All animals were infused with live Escherichia coli, with group I and group II rats additionally receiving a matrix metalloproteinase inhibitor. Serum levels of TNFalpha, interleukin (IL)-6, malondialdehyde (MDA), and lipid hydroperoxide (LOOH) were compared. RESULTS: TNFalpha showed a significant decrease, yet IL-6, MDA, and LOOH (markers of sepsis) levels remained abnormally elevated. CONCLUSION: Despite significantly attenuating TNFalpha, the septic response continued. This supports the concept that in sepsis, monotherapy directed at attenuating a single cytokine cannot overcome the tissue-damaging effects of an entire cascade of mediators. PMID- 10670872 TI - An evaluation of two methods for chronic central venous access device placement. AB - BACKGROUND: Chronic venous access devices (CVADs), placed for phlebotomy and the administration of medications and nutrition, require fluoroscopy to confirm correct catheter position. Long-term central venous catheters placed using an electromagnetic catheter locating system (EMCLS) could result in decreased radiation exposure and decreased cost without compromising accuracy of position. METHODS: Charts of patients who underwent placement of CVADs at University of New Mexico (UNM) Hospital or UNM Cancer Center were reviewed. Inclusion criteria included age >20 years and placement of a central CVAD utilizing fluoroscopy (group 1) or the EMCLS (group 2). Radiation exposure, complications, cost, and accuracy of placement were determined for each technique. RESULTS: Between June 1996 and June 1998, 196 patients underwent placement of CVADs. Complete data sets were available for 46 patients in each group. There were no statistically significant differences in age, gender, complications, or operating room times (P = 0.26). Fluoroscopy and EMCLS were equally accurate for the correct placement of the tip of the line (P = 0.12). Mean patient radiation exposure was EMCLS, 30 mRem, and fluoroscopy, 771 mRem. EMCLS significantly decreased cost (P = 0.025) when compared with fluoroscopic assisted catheter placement. CONCLUSIONS: The use of EMCLS for CVAD placement reduces radiation exposure and cost without compromising the accuracy of placement when compared with standard fluoroscopic assisted placement. PMID- 10670873 TI - Release of anti-inflammatory mediators after major torso trauma correlates with the development of postinjury multiple organ failure. AB - BACKGROUND: Soluble tumor necrosis factor receptor (sTNFr) and interleukin-1 receptor antagonist (IL-1ra) have been identified as endogenous inhibitors of TNF alpha and IL-1beta. While TNF-alpha and IL-1beta levels are not systematically elevated in postinjury patients who developed multiorgan failure (MOF), their involvement at the tissue level has been suggested. Our study hypothesis was that levels of sTNFr-I and IL-1ra would discriminate patients at risk for postinjury MOF. METHODS: Serial plasma levels of sTNFr and IL-1ra were measured in 29 trauma patients at high risk for postinjury MOF. RESULTS: sTNFr-I levels were higher in MOF compared with non-MOF patients at 12, 84, and 132 hours postinjury. MOF patients also had higher IL-1ra values 36, 60, 84, and 132 hours postinjury. CONCLUSIONS: Anti-inflammatory mechanisms are activated after trauma. Since increased levels of sTNFr and IL-1ra correlate with postinjury MOF, they may contribute to our understanding of the pathogenesis as well as prediction of outcome. High levels of antagonists to TNF-alpha and IL-1beta suggest tissue level involvement of these cytokines in postinjury hyperinflammation. PMID- 10670874 TI - Age of transfused blood is an independent risk factor for postinjury multiple organ failure. AB - BACKGROUND: Blood transfusion has repeatedly been demonstrated to be an independent risk factor for postinjury multiple organ failure (MOF). Previously believed to represent a surrogate for shock, packed red blood cell (PRBC) transfusion has recently been shown to result in neutrophil priming and pulmonary endothelial cell activation. We have previously observed that the generation of inflammatory mediators is related to the length of PRBC unit storage. The purpose of this study was to determine if age of transfused PRBC is a risk factor for the development of postinjury MOF. METHODS: Using our prospective database of trauma patients at risk for developing MOF, we identified patients who developed MOF (MOF+) and received 6 to 20 units of PRBCs in the first 12 hours following injury. A similar cohort of patients, matched for ISS and transfusion requirement, who did not develop MOF (MOF-) were also identified. The age of each unit of PRBC transfused in the first 6 hours was determined. Multiple logistic regression was performed to determine if age of transfused blood is an independent risk factor. RESULTS: Sixty-three patients were identified, 23 of whom were MOF+. There was no difference in ISS and transfusion requirement between MOF+ and MOF- groups. MOF+ patients, however, were significantly older (46+/-4.7 years versus 33+/-2.3 years). Moreover, mean age of transfused blood was greater in the MOF+ patients (30.5+/-1.6 days versus 24+/-0.5 days). Similarly, the mean number of units older than 14 and 21 days old were greater in the MOF+ patients. Multivariate analysis identified mean age of blood, number of units older than 14 days, and number of units older than 21 days as independent risk factors for MOF. CONCLUSION: The age of transfused PRBCs transfused in the first 6 hours is an independent risk factor for postinjury MOF. This suggests that current blood bank processing and storage technique should be reexamined. Moreover, fresh blood may be more appropriate for the initial resuscitation of trauma patients requiring transfusion. PMID- 10670875 TI - Attenuation of renal ischemia-reperfusion injury with selectin inhibition in a rabbit model. AB - BACKGROUND: The selectin glycoproteins are involved in the pathogenesis of renal ischemia-reperfusion injury. We investigated the ability of glycyrrhizin, a known selectin inhibitor, to attenuate renal ischemia-reperfusion injury. METHODS: Eighteen New Zealand white rabbits underwent midline laparotomy with renal artery cross-clamping. After 30 minutes of reperfusion, group 1 (control, n = 10) animals received a saline infusion, while group 2 (GLY, n = 8) animals received a glycyrrhizin infusion. Renal function was compared between the two groups after 72 hours of reperfusion. A t test was utilized, with alpha set at P<0.05. RESULTS: Group 1 and group 2 animals had similar baseline renal function. However, after 72 hours of reperfusion, group 1 animals had a significantly higher mean blood urea nitrogen creatinine ratio than group 2 animals (P<0.01), indicating preserved renal function in rabbits treated with glycyrrhizin. CONCLUSIONS: Selectin blockade using glycyrrhizin attenuates renal ischemia reperfusion injury when given 30 minutes after the onset of reperfusion in a rabbit model. PMID- 10670876 TI - Fibrin glue reduces the severity of intra-abdominal adhesions in a rat model. AB - BACKGROUND: The purpose of this study was to determine whether fibrin glue inhibits intra-abdominal adhesions. METHODS: Twenty rats underwent midline laparotomy. To maximize adhesions, bilateral peritoneal muscular defects were created and covered with polypropylene mesh sewn with a braided suture. The bowel was abraded with dry gauze. Rats were randomized to either fibrin glue (FG) sprayed over the mesh or to control (no further treatment) groups. At 1 week, the adhesion density (graded 0 to 3), the percentage of the patch covered by adhesion (0% to 100%), and adhesion type were recorded. RESULTS: The mean adhesion density was 1.45+/-0.33 for FG versus 2.8+/-0.11 for controls (P = 0.001). The mean percentage of adhesions was 36+/-9.9 for the FG group and 94+/-3.7 for controls (P = 0.0002). Bowel or solid organs were adherent to the patch in 6 of 20 (30%) in the FG group versus 12 of 20 (70%) in controls (P = 0.057). CONCLUSIONS: Topical fibrin glue reduces the density and severity of intra-abdominal adhesions in a rat model. PMID- 10670877 TI - Splenectomy in high-risk patients with splenomegaly. AB - BACKGROUND: Splenectomy in patients with massive splenomegaly and hematologic malignancy results in higher morbidity and mortality with primarily palliative benefit. METHODS: From a 14-year experience with 172 splenectomies, the perioperative course of 39 high-risk patients with splenomegaly was reviewed for comorbidities, indications, complications, and mortality. RESULTS: Twenty-three males and 16 females with a mean age of 54.2 years and a mean 12.8-day postoperative length of stay were reviewed. Sixteen patients (41%) had 23 major complications related to age (P = 0.047) and operative time (P = 0.01). Intraoperative transfusion was related to splenic size (P = 0.04), and estimated blood loss (P = 0.02) was inversely related to use of splenic artery preligation. Three perioperative deaths were secondary to sepsis and multi-organ system failure. CONCLUSION: Splenomegaly and comorbidities of the primary disease result in higher morbidity and mortality. Splenic artery preligation is valuable to limit intraoperative blood loss and facilitate splenectomy. PMID- 10670878 TI - Total colectomy versus limited colonic resection for acute lower gastrointestinal bleeding. AB - BACKGROUND: Acute lower gastrointestinal bleeding (ALGB) of the colon can be problematic to diagnose. The purpose of this study was to review our experience with ALGBs and to determine any differences between limited colon resection (LCR) and total/subtotal colon resection (TCR). METHODS: A retrospective study located 77 patients with ALGB, who required 2 or more units of packed red blood cells prior to surgery, and who were taken to the operating room from 1987 to 1997. RESULTS: Fifty LCRs and 27 TCRs were performed during this 10-year period. Recurrent bleeding was significantly more common in the LCR group than in the TCR group (18% versus 4%). Morbidity and mortality were not significantly different. CONCLUSIONS: Owing to the misconception of a higher morbidity with TCR, it has been considered a "last resort" instead of a more expeditious therapy with similar morbidities and mortalities. TCR should be considered more often in the management of these patients. PMID- 10670879 TI - Intraoperative radiofrequency ablation or cryoablation for hepatic malignancies. AB - BACKGROUND: The majority of patients with primary or metastatic malignancies confined to the liver are not candidates for resection because of tumor size, location, multifocality, or inadequate functional hepatic reserve. Cryoablation has become a common treatment in select groups of these patients with unresectable liver tumors. However, hepatic cryoablation is associated with significant morbidity. Radiofrequency ablation (RFA) is a technique that destroys liver tumors in situ by localized application of heat to produce coagulative necrosis. In this study, we compared the complication and early local recurrence rates in patients with unresectable malignant liver tumors treated with either cryoablation or RFA. PATIENTS AND METHODS: Patients with hepatic malignancies were entered into two consecutive prospective, nonrandomized trials. The liver tumors were treated intraoperatively with cryoablation or RFA; intraoperative ultrasonography was used to guide placement of cryoprobes or RFA needles. All patients were followed up postoperatively to assess complications, treatment response, and local recurrence of malignant disease. RESULTS: Cryoablation was performed on 88 tumors in 54 patients, and RFA was used to treat 138 tumors in 92 patients. Treatment-related complications, including 1 postoperative death, occurred in 22 of the 54 patients treated with cryoablation (40.7% complication rate). In contrast, there were no treatment-related deaths and only 3 complications after RFA (3.3% complication rate, P<0.001). With a median follow up of 15 months in both patient groups, tumor has recurred in 3 of 138 lesions treated with RFA (2.2%), versus 12 of 88 tumors treated with cryoablation (13.6%, P<0.01). CONCLUSIONS: RFA is a safe, well-tolerated treatment for patients with unresectable hepatic malignancies. This study indicates that (1) complications occur much less frequently following RFA of liver tumors compared with cryoablation of liver tumors, and (2) early local tumor recurrence is infrequent following RFA. PMID- 10670880 TI - Isoperistaltic bowel lengthening for short bowel syndrome in children. AB - BACKGROUND: Short bowel syndrome, secondary to a variety of causes, can be lethal in infancy and childhood. Isoperistaltic bowel lengthening, performed by longitudinal division of dilated small bowel with end-to-end anastomosis, has shown early promise but long-term outcome is unknown. METHODS: Sixteen infants and children (aged 3 months to 14 years) had short bowel syndrome from necrotizing enterocolitis (8), gastroschisis (4), atresia (2), and volvulus (2). All of these patients were partially or totally dependent on parenteral nutrition and have undergone isoperistaltic bowel lengthening for short bowel syndrome (length <100 cm). Bowel length was increased by 22% to 85% (mean 42%) with the procedure. Studies of intestinal function were performed preoperatively and postoperatively. RESULTS: Isoperistaltic bowel lengthening resulted in significant improvement in stool counts, intestinal transmit time, intestinal clearance of barium, D-xylose absorption, and fat absorption at 6 months and 12 months postoperatively. Fourteen of 16 patients (88%) have been weaned from parenteral nutrition. CONCLUSIONS: These data show that isoperistaltic bowel lengthening can be an effective operation for short bowel syndrome in children, improving absorption and motility, and allowing weaning from parenteral nutrition. PMID- 10670881 TI - Bone substitutes. AB - The search for the ideal bone substitute began hundreds of years ago, and continues today. While numerous choices have been proposed and tested, with varying degrees of success, there remain many challenges related to the use of bone substitutes in craniofacial reconstruction. This paper presents a review of the history of bone substitute research, a discussion of currently popular materials, and elucidation of the challenges to be faced as we approach the new millennium. PMID- 10670882 TI - A new technique for the quantitative analysis of cranial suture biology. AB - OBJECTIVE: Our objective was to assess the ability of the microcomputed tomography scanner to correctly image normal and synostosed cranial sutures at the ultrastructural level. DESIGN AND METHODS: Two specimens of coronal sutures were collected from operative specimens. After appropriate preparation, histological sections were obtained and stained with toluene blue for evaluation. Representative histological sections were compared to microcomputed tomography slices. RESULTS AND CONCLUSIONS: With microcomputed tomography, we successfully imaged one normal and one synostosed human coronal suture and performed a quantitative analysis of these specimens. Microcomputed tomography scanning was found to be a highly accurate imaging device for the evaluation of cranial suture development. Microcomputed tomography offers three-dimensional imaging at the microscopic level and allows for rapid quantitative analysis of bone architecture, including several measurements unavailable through histologic analysis. We believe that microcomputed tomography can play an important role in imaging and in the quantitative analysis of the stereology of bone microarchitecture. Among its advantages, microcomputed tomography is able to image many more slices than are obtainable through histology, and the method is not prone to human error. Microcomputed tomography slices are generated without destruction of the specimen and without loss or corruption of reproducible data. Structure-oriented slices from microcomputed tomography together with cellular oriented sections from histology are complementary in the overall quantitative analysis of cranial sutures. PMID- 10670883 TI - Dentoalveolar relations in children born with a unilateral cleft lip and palate (UCLP) in Western Australia. AB - OBJECTIVE: Our objective was to evaluate complete unilateral cleft lip and palate repair outcome in the Cleft Unit in Perth, Western Australia, by assessment of dentoalveolar relationships. DESIGN: This is a retrospective study. SETTING: Our subjects were individuals under the care of the cleft team in Perth, Western Australia. PARTICIPANTS: All patients with unilateral cleft lip and palate and available 6-year casts who had been born since January 1, 1985, were identified from the cleft unit's database. The nature of the cleft was verified by examination of birth study models and photographs. A total of 54 such patients were identified. MAIN OUTCOME MEASURES: Main outcome measures were identified through dental arch relationship grading of study models using the 5 Year Old Study Model Index. RESULTS: Interexaminer and intraexaminer agreement kappa statistics revealed good to very good agreement using this index. The results indicate that the surgical outcome was graded as excellent, good, or fair for 77% of patients and poor or very poor for 23% of patients. CONCLUSIONS: The results of the Western Australia study compare favorably to the overall U.K. outcome (the Clinical Standards Advisory Group study) but unfavorably to the results of some European centers, such as Oslo. PMID- 10670884 TI - The adult unoperated cleft patient: absence of maxillary teeth outside the cleft area. AB - OBJECTIVE: The purpose of this study was to investigate the possible absence of teeth in the postcanine region of the upper jaw of the unoperated adult cleft patient. METHOD: The study was performed on 266 dental casts of fully unoperated adult cleft patients. The patients were divided into four groups according to the type of the cleft: unilateral cleft lip and alveolus, unilateral cleft lip and palate, bilateral cleft lip and alveolus, and bilateral cleft lip and palate. RESULTS: No absence of permanent teeth in the canine and postcanine area of the upper jaw could be found. CONCLUSION: The results are in contradiction with the established hypothesis that absence of teeth outside the cleft area of the maxilla is due to an unknown congenital factor. On the contrary, the findings support the hypothesis that surgery for the closure of the hard palate in early childhood is the most important etiological factor for the absence of teeth outside the cleft area in the early operated cleft patient. The superficial position of the tooth germs (at the time of the palatal surgery), especially those of the premolars, supports this hypothesis. PMID- 10670885 TI - Radiographic determination of the position of the maxillary lateral incisor in the cleft alveolus and parameters for assessing its habilitation prospects. AB - OBJECTIVE: Develop patient selection criteria and guidelines for the diagnostic work-up for early secondary alveolar bone graft designed to accommodate a lateral incisor in the cleft. DESIGN: A literature review with clinical examples. INTERVENTIONS: For the majority of cleft lip and palate (CLP) patients, the timing of the secondary alveolar bone graft procedure is devised to accommodate the eruption of the maxillary canine. However, when the lateral incisor is present in the cleft area, a decision may be made to perform early grafting in order to salvage an additional tooth. Appropriate case selection for this intervention is the key for successful dental habilitation in the cleft area. RESULTS AND CONCLUSIONS: Attempts to maintain the lateral incisor are recommended only if the tooth contributes to the comprehensive orthodontic and restorative patient care and when the presence of a lateral incisor with adequate size, shape, and proper position may be demonstrated. For optimal decision making, the position, form, and periodontal ligament support of the lateral incisor should be assessed. Evaluation of these parameters is based chiefly on radiographic imaging. Occlusal projections and traditional tomography may augment the information available from periapical and panoramic radiographs. However, each of these has benefits and drawbacks that require consideration. PMID- 10670886 TI - The basis for presurgical orthopedic treatment of infants with unilateral complete cleft lip and palate. AB - OBJECTIVE: The purpose of this study was to describe the posttreatment morphology of the upper part of the oral cavity of complete unilateral cleft lip and palate (UCLP) patients and to compare it to noncleft contemporaries. Patients were treated according to a protocol designed to keep a proper resting posture of the oral cavity. DESIGN: Retrospective study on dental casts. SETTING: The study was performed at a maxillofacial center serving a population of 2 million inhabitants. Data for noncleft subjects are the result of a longitudinal study at the same institution. PATIENTS: Twenty-one Caucasian UCLP patients (13 males, 8 females) aged 5 to 9 years with no other congenital anomalies and no postoperative orthodontic treatment. INTERVENTIONS: Each patient received 5 to 6 months of preoperative orthopedics with a passive plate and external nonelastic strapping with definitive lip repair at age 5 to 7 months, soft palate repair at 11 to 15 months, and hard palate repair with mucoperiosteal closure of the alveolus at 30 to 36 months. Each patient was compared to the mean values obtained from a longitudinal study of a group of 25 healthy noncleft children of the same ethnic group (11 males, 14 females). RESULTS: Analysis of dental casts indicated that 16 patients had a width, depth, and length of the alveolar arch in the range of the mean normal values minus two standard deviations. Their analyzed palates were flatter than normal. Six of 21 children had too small an alveolar arch for their ages, and they did not acquire a correct posture of the oral cavity. CONCLUSION: The results indicate that the upper part of the oral cavity of UCLP patients can reach the dimensions of noncleft contemporaries despite surgery. PMID- 10670887 TI - Genetics of nonsyndromic cleft lip and palate: a review of international studies and data regarding the Italian population. AB - The aims of this review are (1) to illustrate current knowledge of the mode of inheritance and the loci involved in the cleft lip and palate and (2) to summarize the results of our investigations, which were carried out in Italy. It is well established that nonsyndromic cleft of the lip with or without the palate (CL+/-P) and cleft palate only (CPO) are separate entities. Genetic heterogeneity has been observed in CL+/-P, which involves different chromosome regions, mainly 6p23 (OFC1), 2q13 (OFC2), and 19q13.2 (OFC3), as well as other loci, such as 4q25 4q31.3 and 17q21. Furthermore, an interaction between different genes has been suggested in the oligogenic model. In one case at least, an OFC1 and OFC2 interaction has been demonstrated. The mode of inheritance of CPO is compatible with a recessive single major gene model, while an association with a candidate gene, mapping on the chromosome region 2q13/TGFalpha, remains to be confirmed. PMID- 10670888 TI - Associated malformations in cases with oral clefts. AB - OBJECTIVE: Infants with oral clefts (OCs) often have other associated congenital defects. The reported incidence and the types of associated malformations vary between different studies. The purpose of this investigation was to assess the prevalence of associated malformations in a geographically defined population. METHOD: The prevalences at birth of associated malformations in infants with OCs were collected between 1979 and 1996 on all infants born in the area covered by the registry of congenital anomalies of Northeastern France in 238,942 consecutive births. RESULTS: Of the 460 cleft infants born during this period, 36.7% had associated malformations. Associated malformations were more frequent in infants who had cleft palate (46.7%) than in infants with cleft lip and palate (36.8%) or infants with isolated cleft lip (13.6%). Malformations in the central nervous system and in the skeletal system were the most common other anomalies, followed by malformations in the urogenital and cardiovascular systems. Weight, length, and head circumference of children with OCs and multiple associated malformations were lower than in controls, as was the weight of the placenta. Prenatal diagnosis was rarely done by fetal ultrasonographic examination in isolated clefts. However, even in multiple associated malformations, prenatal diagnosis by fetal ultrasonographic examination had a low sensitivity, 31.6%. CONCLUSION: The overall prevalence of malformations, which was one in more than three infants, emphasizes the need for a thorough investigation of infants with clefts. A routine screening for other malformations especially skeletal, central nervous system, and cardiac defects may need to be considered in infants with clefts, and genetic counseling seems warranted in most of these complicated cases. PMID- 10670889 TI - A three-dimensional quantitative analysis of lips in normal young adults. AB - OBJECTIVE: To supply information about (1) sex-related dimensions (linear distances and ratios, vermilion area, volume) of normal adult lips, (2) presence of sexual dimorphism, and (3) correlations between anthropometric characteristics of the lip and nose. METHODS: The three-dimensional coordinates of soft tissue landmarks on the lips and nose were obtained using an optoelectronic instrument in 90 healthy young adult women and 90 healthy young adult men. From the landmarks, several linear distances (mouth width, total vermilion height, nose height, anatomic nose width, total lip height, upper lip height), the ratio of vermilion height to mouth width, and some areas (vermilion of the upper lip, vermilion of the lower lip, total vermilion) and volumes (upper lip volume, lower lip volume, total lip volume) were calculated. Linear correlation analyses between pairs of variables were also conducted within each sex. RESULTS: All lip dimensions (distances, areas, and volumes) were significantly larger in men than in women (p<.005), but no sex differences were found in the vermilion height to mouth width ratio. Overall, mouth and nose dimensions were not significantly correlated, with the exceptions of the upper and lower lip volumes in both sexes and of the mouth and nose widths in the female sample, in which a modest part of the variance in one measurement could be explained by the other. CONCLUSION: The dimensions of the mouth and the nose did not seem to be strictly related. Data collected in the present investigation could represent a database for the quantitative description of human lip morphology in adult subjects. PMID- 10670890 TI - Histological evaluation of autogenous iliac particulate cancellous bone and marrow grafted to alveolar clefts--a preliminary report of five young adult cases. AB - OBJECTIVE: We histologically evaluated iliac particulate cancellous bone and marrow (PCBM) grafted to alveolar clefts in five young adults with cleft lip and palate. PATIENTS AND METHODS: Five young adults with cleft lip and palate underwent secondary alveolar bone grafting. The mean age of the patients was 21.2 years (range = 17 to 27 years). Bone specimens were taken from the graft site 5 to 10 months after the surgery simultaneously with implant fixture placement (two cases), vestibuloplasty (two cases), and rebone grafting (one case). These five bone specimens were prepared and stained with hematoxylin and eosin and examined microscopically. RESULT: One specimen, taken 5 months after surgery, showed immature trabecular bone partially lined by osteoblasts. The remaining four specimens showed well-mineralized trabeculae and fatty change in the marrow space. CONCLUSION: In young adult cases, PCBM remodeled after grafting and became mature bone about 5 to 6 months after the surgery. PMID- 10670891 TI - Laryngeal airway resistance in cleft palate children with complete and incomplete velopharyngeal closure. AB - OBJECTIVE: This study investigated the effect of velopharyngeal insufficiency on aerodynamic measures of laryngeal function in children with cleft palate. DESIGN: Data were analyzed using analysis of covariance. The independent variable was velopharyngeal closure, and the dependent variables were laryngeal resistance, laryngeal airflow, and transglottal pressure. Age and gender were covariates. SETTING: The data were collected at The Craniofacial Center, University of Illinois, a tertiary health care center located in Chicago. PATIENTS: Thirty-six children with cleft palate were recruited from among the patients at The Craniofacial Center. Ten children with velopharyngeal areas >5 mm2 during oral speech were placed in the incomplete closure group, while 26 children with areas <1 mm2 were placed in the complete closure group. OUTCOME MEASURES: The three dependent variables (transglottal pressure, transglottal airflow, and laryngeal resistance) were measured. RESULTS: Laryngeal resistance and transglottal pressure were significantly higher, and transglottal airflow was significantly lower in the group with complete closure. CONCLUSIONS: In summary, cleft palate patients with complete velopharyngeal closure exhibited higher laryngeal resistances than those with incomplete closure. PMID- 10670892 TI - Mental development in infants with cleft lip and/or palate. AB - OBJECTIVE: Investigated mental development in infants and toddlers with cleft lip and/or palate (CLP). DESIGN: This was a retrospective analysis of developmental scores on qualified children between 4 and 36 months of age. Cross-sectional analysis included children in four age groups (6, 12, 18, and 24 months); longitudinal analysis included children at mean age 9.1 (range = 4 to 15) months at Time 1 and 24 months (range = 16 to 36) at Time 2. PARTICIPANTS: Cross sectional analysis included 180 children (59% male participants) in four diagnostic groups (cleft lip only [CL], cleft lip and palate [CLP], cleft palate only [CP], and Pierre Robin). The longitudinal sample included 85 children (64% male children) in the same diagnostic groups. MAIN OUTCOME MEASURES: Mental Scale (MDI) of the Bayley Scales of Infant Development. RESULTS: Mean MDIs were in the average range but decreased significantly between youngest and oldest groups in both cross-sectional (F(3,179) = 4.9, p<.01) and longitudinal samples (F(1,84) = 6.87, p<.01). There was a significant difference among cleft types (F(3,179) = 3.5, p<.025). Infants with CL obtained the highest scores, and infants with Pierre Robin Sequence obtained the lowest. Perceptual-motor development in the first year of life was predictive of developmental status at age 2. CONCLUSIONS: The number of children with CLP who may be at risk for developmental problems during the second year of life is greater than would be expected. Children at greatest risk may demonstrate early delays in acquisition of perceptual-motor skills during the first year of life. PMID- 10670893 TI - Facial growth in a cleft palate patient treated with the Herbst appliance: a long term profile roentgenographic and roentgen stereometric analysis of profile changes and displacement of the jaws. AB - OBJECTIVE: To monitor facial development in a patient with cleft palate who was treated with the Herbst appliance. Monitoring was in terms of changes in the skeletal profile and growth in the circummaxillary sutures and temporomandibular joints (TMJs). DESIGN: Prospective profile roentgenography (between the ages of 6 and 20 years) and roentgen stereometric analysis (between the ages of 8 and 19 years). SETTING: Center for Craniofacial Anomalies and Department of Plastic and Reconstructive Surgery, Malmo University Hospital, Malmo, Sweden. PATIENT: Boy with cleft of the soft and posterior part of the hard palate and marked facial convexity. INTERVENTIONS: Surgical repair of the soft palate at age 9 months, velopharyngeal flap at age 8 years, and insertion of implants under general anesthesia and treatment with the Herbst appliance at age 11 years. Roentgen examinations were performed in connection with continued clinical evaluations and treatment. MAIN OUTCOME MEASURES: Profile roentgenograms were traced and measured by one of the authors using conventional point-based analysis. Stereo roentgenograms were digitized by the Department of Orthopedic Surgery, Malmo University Hospital. RESULTS: The direction of profile changes was partly discordant with the direction of articular growth in the circummaxillary sutures and TMJs. The successful treatment result was accomplished by a temporary influence on sagittal growth direction in the circummaxillary sutures and on rotational growth direction in the TMJs, combined with a favorable natural remodeling and articular growth pattern. CONCLUSIONS: The mode of growth by which treatment aims were reached was partly unexpected, i.e., discordant with the generally accepted principal concept that treatment with the Herbst appliance positions the mandible forward. PMID- 10670894 TI - Crouzon syndrome with acanthosis nigricans: case report and mutational analysis. AB - OBJECTIVE: To describe the 22nd case of Crouzan syndrome with acanthosis nigricans, a hyperkeratotic skin disorder with hyperpigmentation. METHODS: DNA analysis and sequencing of the FGFR3 gene were performed. RESULTS: The 13-year old Japanese boy described here also had dyspnea, facial palsy, sensorineural hearing loss, and skeletal and mental retardation. Examination of a skin biopsy specimen revealed the typical findings of acanthosis nigricans. Genetic analysis revealed the Ala391Glu mutation in one FGFR3 gene. CONCLUSIONS: Crouzon syndrome with acanthosis nigricans is a distinct clinical entity different from classic Crouzon syndrome. PMID- 10670895 TI - Epignathus teratoma: report of three cases with a review of the literature. AB - Three cases of epignathus teratoma associated with other midline anomalies are reported. The first case involved Pierre Robin sequence and a bifid tongue. The second case was characterized by two teratomas, a meningoencephalocele, and a cleft lip and nose. The third case had Pierre Robin sequence associated with duplication of the pituitary gland and hypoplasia of the corpus callosum. PMID- 10670896 TI - Effects of maxillary protraction combined with chin-cap therapy in unilateral cleft lip and palate patients. AB - OBJECTIVE: This study investigated the treatment effects of maxillary protraction combined with chin-cap therapy in complete unilateral cleft lip and palate patients at the deciduous and early mixed dentition stages. METHOD: Twenty-six Japanese children (10 boys and 16 girls) with complete unilateral cleft lip and palate were examined. All had undergone pushback operations for palatal repair at approximately 18 months of age. Maxillary protraction began between 5 and 7 years of age and continued for 10 to 38 months. Lateral cephalograms were used to analyze skeletal changes during the first and second years of treatment, and the relationship between pretreatment midfacial morphology and forward displacement of the maxilla during the first year was investigated. RESULTS AND CONCLUSIONS: During the first year of treatment, the mean increase in the ANB angle for all cases was 2.37 degrees. The forward displacement of the maxilla varied considerably, from 0.23 mm to 3.03 mm. The treatment response was significantly smaller in the second year, and no benefit from treatment longer than 1 year was established. The amount of maxillary forward displacement was significantly correlated with the pretreatment posterior upper facial height. Patients with smaller posterior upper facial height showed a poorer treatment response, whereas patients with a greater posterior upper facial height responded better to treatment. Individual differences in maxillary growth acceleration may be related to growth inhibition associated with postsurgical scar tissue on the palates. PMID- 10670897 TI - Combined bone grafting and delayed closure of the hard palate in patients with unilateral cleft lip and palate: facilitation of lateral incisor eruption and evaluation of indicators for timing of the procedure. AB - OBJECTIVE: To compare outcomes of bone grafting performed before eruption of the lateral incisor to outcomes of grafting performed before eruption of the canine and to evaluate the long-term results of bone grafting combined with delayed closure of the hard palate during mixed dentition. DESIGN: Seventy consecutive patients (52 men and 18 women) with complete unilateral cleft lip and palate were studied. All patients underwent bone grafting with simultaneous closure of the cleft in the hard palate at the stage of mixed dentition. The velum had been repaired in infancy. Mean age for the bone grafting procedure was 8.4 years. Bone grafting was performed to facilitate eruption of the lateral incisor in 43 (61%) of the patients and to facilitate eruption of the canine in the remaining 27 (39%) patients. Intraoral radiographs were used to evaluate the morphologic characteristics of the cleft and the stage of eruption of the permanent lateral incisor and canine before bone grafting. Mean follow-up time was 4.0 years (range, 1-10.1 years). RESULTS: The mean time for the surgery, which included bone grafting and repair of the residual cleft in the hard palate, was 109 minutes, and the mean amount of bleeding was 121 ml. The rate of dehiscence in the flap covering the alveolar bone graft was 14%, and the rate of total failure of bone grafting was 3%. An oronasal fistula developed in the hard palate of 13% of patients, but the fistula was of sufficient size to serve as an indication for reoperation in only 6%. The postoperative alveolar bony height in the cleft area was more than 75% of the normal height in 94% of patients. Closure of the cleft space in the dental arch was performed or planned to be achieved orthodontically in 91% of patients. When bone grafting was performed to facilitate eruption of the lateral incisor, the cleft space was closed orthodontically in 100% of patients. The optimal indicator for timing of the bone grafting procedure from an orthodontic point of view was when the permanent lateral incisor or the canine close to the cleft was covered by a thin shell of bone (i.e., 7-9 years of age). PMID- 10670898 TI - NMR determination of dynamic parameters of CH3 groups in P(CH3)4SbCl6. AB - The longitudinal relaxation time T1 and the second moment M2 of 1H NMR line in a wide temperature range have been measured for P(CH3)4SbCl6. It was found that two different methyl groups in each tetramethylphosphonium cation perform two different rates of C3 motions. The reduction of the proton second moment M2 just below the temperature of the phase transition Tc2 = 350 K may suggest that the isotropic tumbling of the whole cation [P(CH3)4]+ is involved in the structural change of the crystal lattice induced by the movements of the [SbCl6]- anion. PMID- 10670899 TI - CP-MAS 207Pb with 19F decoupling NMR spectroscopy: medium range investigation in fluoride materials. AB - The isotropic chemical shift of 207Pb is used to perform structural investigations of crystalline fluoride compounds (PbF2, Pb2ZnF6, PbGaF5, Pb3Ga2F12 and Pb9Ga2F24) and transition metal fluoride glasses (TMFG) of the PZG family (PbF2-ZnF2-GaF3). Using 207Pb Cross Polarisation Magic Angle Spinning (CP MAS) NMR with 19F decoupling, it is shown that the isotropic chemical shift of 207Pb varies on a large scale (1000 ppm) and that the main changes of its value are not due to the nearest neighbour fluorines but may be related to the number of next nearest neighbour (nnn) Pb2+ ions. In this way, it is demonstrated that 207Pb chemical shift is an interesting probe to investigate medium range order in either crystalline or glassy fluoride systems. The 207Pb delta(iso) parameter has been linearly correlated to the number of nnn Pb2+ ions. PMID- 10670900 TI - Proton magnetic resonance microimaging of human trabecular bone. AB - Proton magnetic resonance (1H magnetic resonance imaging (MRI)) images of human trabecular bone were acquired and discussed for two samples with different porosity. Three-dimensional 3D Spin Echo (3D SE) and Multi-Slice Multi-Echo (MSME) pulse sequences were examined. A very high slice resolution of (38 microm)2 was achieved (MSME). The intensity histograms were found useful for the characterization of the bone porosity. A spatial distribution of the spin-spin relaxation time T2 was monitored with the MSME pulse program. The work demonstrates the great potential of the proton MRI technique in the study of the trabecular bone morphology. PMID- 10670901 TI - NMR microscopy of polyurethane foams. AB - NMR microimaging has been applied to the characterization of foam materials, in order to understand their properties and gain deeper insight into their structures. The structure of "open cell" foams can be visualized with sufficient resolution to obtain the size and shape of cells, as well as their spatial distribution within specimens. Statistics of cell sizes have been calculated from NMR microscopy and are in good agreement with results obtained from electron microscopy. We demonstrate that the local density of foams, which are parameters largely inaccessible by other analytical methods, readily can be determined by NMR microscopy. PMID- 10670902 TI - 207Pb NMR of minium, Pb3O4: evidence for the [Pb2]4+ ion and possible relativistic effects in the Pb-Pb bond. AB - Solid Pb3O4 has been studied with 207Pb nuclear magnetic resonance (NMR) spectroscopy. The 207Pb NMR chemical-shift tensor of the Pb2+ site has principal values of delta11 = 1980 +/- 5 ppm, delta22 = 1540 +/- 5 ppm, and delta33 = -1108 +/- 10 ppm; delta(iso) = 804 +/- 10 ppm. The chemical-shift tensor of the Pb4+ site is axial, with principal values delta(parallel) = -1009 +/- 3 ppm and delta(perpendicular) = 1132 +/- 3 ppm; delta(iso) = -1091 +/- 3 ppm. The Pb4+ Pb2+ scalar coupling constant J(Pb-Pb) = 2.3 +/- 0.1 kHz. The main contribution to the Pb2- chemical-shift anisotropy is proposed to arise from an exchange interaction in the Pb2+-Pb2+ pairs, conventionally regarded as molecular [Pb2]4+ ions. PMID- 10670903 TI - 6Li and 7li solid-state NMR spectroscopy of nitrogen ceramic phases. AB - Two nitrogen ceramic phases, the oxynitride LiSiON and the nitride LiSi2N3, have been studied by 6Li and 7Li NMR. Magic angle spinning (MAS) NMR experiments have been carried out at two magnetic field strengths (7.05 and 14.1 T). The spectra give evidence of the relative effects of the quadrupolar and chemical shift interactions. The electric field gradient tensor of both phases has been determined accurately by iterative fitting of the 6Li and 7Li MAS NMR line shapes at the two magnetic field strengths. Due to the fact that for 7Li the quadrupolar interaction is much larger than the chemical shift interaction, it is shown that neither the small chemical shift anisotropy nor the relative orientation of the two interaction tensors can be determined accurately by 7Li MAS NMR. For 6Li, the two interactions are comparable and the value of these parameters obtained from the fits of the 6Li experimental MAS line shapes are therefore much more reliable. PMID- 10670904 TI - 207Pb chemical shift thermometer at high temperature for magic angle spinning experiments. AB - The temperature dependence of 207Pb chemical shift in magic angle spinning (MAS) NMR spectrum of Pb(NO3)2 provides a sensitive method to calibrate sample temperatures in MAS NMR. The temperature dependence is uniform in the temperature range between 30 degrees C and 400 degrees C. The NMR sensitivity and the line width are also favorable. PMID- 10670905 TI - From crystalline to glassy gallium fluoride materials: an NMR study of 69Ga and 71Ga quadrupolar nuclei. AB - Owing to the implementation of acquisition techniques specific for nuclei with very large quadrupolar interaction (full shifted echo and variable offset cumulative spectra (VOCS)), NMR spectra of 69Ga and 71Ga are obtained in crystallised (PbGaF5, Pb3Ga2F12, Pb9Ga2F24 and CsZnGaF6) and glassy (PbF2-ZnF2 GaF3) gallium fluorides. Simulations of both static (full echo or VOCS) and 15 kHz MAS spectra allow to obtain consistent determinations of isotropic chemical shifts and very large quadrupolar parameters (nuQ up to 14 MHz). In the crystalline compounds whose structures are unknown, the number and the local symmetry of the different gallium sites are tentatively worked out. For the glassy systems, a continuous Czjzek's distribution of the NMR quadrupolar parameters accounts for the particular shape of the NMR spectrum. PMID- 10670906 TI - Nicotinamide inhibits enhanced in vitro production of interleukin-12 and tumour necrosis factor-alpha in peripheral whole blood of people at high risk of developing type 1 diabetes and people with newly diagnosed type 1 diabetes. AB - Macrophages and T lymphocytes are the first cells to appear in pancreatic islets in the development of autoimmune diabetes. It has been suggested that cytokines released by monocytes/macrophages, including interleukin-1beta (IL-1beta), interleukin-12 (IL-12) and tumour necrosis factor-alpha (TNF-alpha) could have an initial role in islet B-cell damage. The aim of the present study was to estimate the effect of human insulin and nicotinamide on the levels of monocyte/ macrophage derived cytokines in the peripheral blood of humans at risk of Type 1 diabetes, and in patients with newly diagnosed Type 1 diabetes compared to healthy control subjects. The study was carried out on three groups of subjects: 20 first degree relatives of people with Type 1 diabetes (with two or more antibodies against pancreatic B-cell antigens); 22 patients with recent onset of Type 1 diabetes (duration of the disease 3-6 months); and 25 age- and sex-matched healthy subjects. Cytokine levels (IL-1beta, IL-12, and TNF-alpha) in the supernatants of whole blood cultures incubated with PHA alone (10 microg/ml), or PHA + human insulin (50 microg/ml), or PHA + nicotinamide (100 micromol/l) were quantified by ELISA. In the cultures with nicotinamide the concentration of IL-12 and TNF-alpha was significantly lower in the prediabetic group, diabetic patients, and the healthy controls than in the cultures with PHA only or with PHA + insulin. There were no significant differences in IL-1beta production in the cultures after incubation with the different stimuli in the studied groups and healthy controls. No significant influence of human insulin on macrophage/monocyte cytokines secretion in in vitro cultures of the peripheral blood was found. This suggests that nicotinamide could influence monocyte/macrophage function in peripheral blood by inhibiting production of IL 12 and TNF-alpha. PMID- 10670907 TI - Methods for assessing diabetic polyneuropathy: validity and reproducibility of the measurement of sensory symptom severity and nerve function tests. AB - The usefulness of sensory symptoms in the assessment of diabetic polyneuropathy is unclear. In the present study, we studied the hypothesis that pain is associated with small nerve fibre function, and that sensory alteration is associated with large nerve fibre function. In addition, we assessed the reproducibility and the ability to detect changes in clinical status over time of the nerve function tests currently used in clinical trials. Patients (78) with stable diabetic polyneuropathy were examined on three separate occasions with a test-retest interval of 17 and 52 weeks. Small nerve fibre function was measured using temperature discrimination thresholds for warmth (TDTwarmth) and cold (TDTcold). Large nerve fibre function was measured by testing sensory and motor nerve conduction velocities (SNCV and MNCV) and vibration perception thresholds (VPT). Neuropathic pain was only significantly associated with TDTcold, and with the MNCV of the tibial nerve. Sensory alteration was associated with almost all nerve function tests except the SNCV and MNCV of the ulnar nerve. The measurements of symptom severity and the nerve function tests all proved to be sufficiently reproducible. The standardized smallest detectable difference on group level (SDD) of the measurement of sensory alteration and neuropathic pain were almost the same (9% and 12%, respectively). Among the nerve function tests, the SNCV and MNCV had the smallest SDD (3-4%), and were, therefore, potentially the most responsive instruments. The SDD of the TDT was greater than the VPT (9 14% vs 21-28%, respectively). In conclusion, neuropathic pain was not associated with small nerve fibre function, and sensory alteration was associated with both large and small fibre function. In addition, the standardized measurement of symptom severity, the SNCV and MNCV tests, and the VPT test appear to be useful for monitoring the course of polyneuropathy in clinical trials. PMID- 10670908 TI - Delapril versus manidipine in hypertensive therapy to halt the type-2-diabetes mellitus-associated nephropathy. AB - Thirty-nine hypertensive patients with type 2 diabetes mellitus were followed under long-term treatment (mean, 20.7 months) with manidipine hydrochloride, a Ca antagonist, or delapril hydrochloride, an ACE inhibitor, at nine institutions. Both the treatments showed similar antihypertensive effects, although slight but significantly larger decreases were observed in systolic and mean blood pressures at months 12 and 24 in the patients treated with manidipine (P < 0.02). The urinary albumin excretion index (AEI) tended to increase throughout the study in both treatment groups, but no significant difference in AEI was observed between the two treatment groups at any time point. Overt albuminuria developed in four patients on manidipine but did not appear in any of the patients on delapril. The risk of progression to overt albuminuria was significantly different between manidipine and delapril groups (P = 0.011). No increase in serum creatinine (Cr) was observed with delapril. The average excretion indexes of tubular markers such as beta2-microglobulin, alpha1-microglobulin, and NAG tended to be higher in the patients on manidipine than in those on delapril. Taken in sum, these findings suggest that the ACE inhibitor delapril is more beneficial than the Ca antagonist manidipine in the treatment of diabetic renal diseases via mechanisms other than the blood pressure regulation, partly through their different effects on tubular function. In conclusion, delapril was significantly more effective than manidipine in inhibiting progression to overt albuminuria in hypertensive type 2 diabetes mellitus patients. PMID- 10670909 TI - Normal pressure hydrocephalus in diabetic patients with recurrent episodes of hypoglycemic coma. AB - The pathophysiology of brain damage induced by severe hypoglycemia is still unknown. We experienced a case with type 1 diabetes and recurrent severe hypoglycemic coma who showed a central brain atrophy and an abnormal cerebrospinal fluid flow, suggesting normal pressure hydrocephalus. Following this case, the CSF flow was studied using 111In-DTPA cisternography in six consecutive diabetic patients admitted for repeated episodes of hypoglycemic coma. All the patients showed the central brain atrophy on computed tomography and four of them (67%) had the ventricular reflux, with delayed clearance of 111In-DTPA. Two patients with abnormal CSF flow showed cognitive dysfunction by WAIS or WAIS-R. In contrast, none of five randomly selected diabetic patients, without hypoglycemic coma showed abnormal CSF flow. Our results suggest the presence of normal pressure hydrocephalus in diabetic patients with recurrent hypoglycemic coma. It may associate with the cognitive dysfunction. PMID- 10670910 TI - Out-patient management does not impair outcome of pregnancy in women with type 1 diabetes. AB - In recent years, out-patient protocols have mainly displaced historical obstetric management of diabetic pregnancy. The impact of the change from centralized in patient to decentralized out-patient treatment on glycaemic control and its effects on the outcome of newborns in diabetic pregnancies was therefore studied using the population-based data on 296 pregnancies in 224 women with type 1 diabetes over 10 years (1986-1995) in the two northernmost provinces of Finland. The area comprises one tertiary level and four other central hospitals. The change of policy was effected in 1990 and to determine the impact of this change, the study period was divided in two (period 1, 1986-1990, n = 135; period 2, 1991 1995, n = 161). At the first antenatal contact (mean 9.9 weeks of gestation) 73% of women had unsatisfactory glycaemic control, but it improved rapidly with pregnancy and was significantly better (P < 0.05) in the second study period. The incidence of congenital malformations was somewhat greater (NS) in period 2 but perinatal mortality did not change. Out-patient management does not impair outcome in type 1 diabetic pregnancy. PMID- 10670911 TI - Serum sialic acid in young type-1 diabetic patients. AB - It is well established that serum total sialic acid (TSA) is elevated in patients with type-2 diabetes mellitus (NIDDM) compared to non-diabetics. However, it is not clear whether serum TSA is also elevated in type-1 diabetic patients (IDDM). Twenty-one type-1 patients were studied along with age and sex matched normal non diabetic subjects (ten males and 11 females). Their ages were 24.8+/-3.4 years (20-30) and 23.5+/-3.9 years (18-30) respectively. The duration of diabetes mellitus was 12.6+/-6.7 years (1-24) with a HBA1c of 9.0+/-2.2% (6.0-14.9). There was no significant difference in serum TSA of the type-1 diabetic patients 689+/ 107 mg/l versus 670+/-119 mg/l in the normal subjects. Nor was there a significant correlation between serum TSA with patient age (r = -0.31), urine albumin/creatinine ratio (ACR) (r = 0.25), HBA1c (r = 0.36), plasma random glucose (r = -0.04) or diabetes duration (r = -0.09) in the diabetic patients. However, there was a significant correlation between serum TSA and mean daily insulin dose (r = 0.51, P<0.02) and also serum cholesterol and triglyceride (r = 0.58, P<0.01 and r = 0.49, P<0.04, respectively) in the type-1 diabetic patients. In summary, we conclude that serum TSA is not elevated in young type-1 diabetic patients compared with normal age and sex matched control subjects. However, the relationship between serum TSA and serum lipids and also mean daily insulin dose merits further research. PMID- 10670912 TI - Painful diabetic polyneuropathy: epidemiology, pain description, and quality of life. AB - A prospective survey study was performed in patients with painful diabetic polyneuropathy (PDN) to assess the nature and scope of their pain. Pain associated with diabetic neuropathy is commonly encountered in clinical practice. Yet, little is known regarding the pain experience and impact on quality of life in persons with painful diabetic neuropathy. These 105 patients noted an average of 6/10 pain, most often described as 'burning', 'electric', 'sharp', and 'dull/ache', which, for most, is worse at night time and when tired or stressed. On average, patients reported that the pain caused substantial interference in sleep and enjoyment of life and moderate interference in recreational activities, normal work, mobility, general activity, social activities, and mood. Unexpectedly, a potential genetic predisposition to the development of painful neuropathy was suggested by the fact that a majority (56%) reported a family member with PDN. Thus, this study found that pain associated with diabetic neuropathy is a significant medical issue that has a substantial impact on the quality of life of many people with this condition. PMID- 10670913 TI - Limitations of glycosylated haemoglobin as an index of glucose intolerance. AB - This study was conducted (a) to establish a normal cut-off value for glycosylated haemoglobin measured as HbA1c in South Indian subjects, and (b) to evaluate its usefulness in demarcating different categories of glucose intolerance. HbA1c measurement was carried out in 1261 cases with no known history of diabetes, while being tested by oral glucose tolerance test (M:F 850:411, mean age 40+/-12 years). An immunoturbidimetric procedure for HbA1c assay (Tina-Quant, Boehringer Mannheim, Germany) was used. The specificity and sensitivity of HbA1c in demarcating normal glucose tolerance (NGT) from abnormal tolerance were calculated using the ROC procedure. By the ROC analysis, a cut-off value of HbA1c > or = 6.0% gave a sensitivity of 88.5% and specificity of 62.8% using the WHO criteria (2-h plasma glucose > or = 200 mg/dl). Using the ADA criterion (fasting plasma glucose > 125 mg/dl) the sensitivity and specificity for the same cut-off value were 85.2 and 61.2%. In NGT, only a small percentage of the variance in HbA1c was explained by the fasting plasma glucose (FPG) values. The overall correlation coefficient between the fasting plasma glucose and HbA1c was r = 0.8, r2 = 0.64 and, in the case of 2-h post glucose, r = 0.82, r2 = 0.67. This showed that more than 35% of the variations in HbA1c were not explained by the plasma glucose values. The study showed that HbA1c values of > or = 6.0% gave a reasonably high sensitivity and specificity for diagnosis using the WHO or ADA criteria. However, nearly 35% of the variations in HbA1c were not explained by the variations in plasma glucose. Wide inter-individual variations even in the normoglycaemic range make the test unsuitable for diagnostic purpose. PMID- 10670914 TI - Prevalence of type 2 diabetes mellitus and impaired glucose tolerance in the Kashmir Valley of the Indian subcontinent. AB - This cross-sectional population survey was undertaken to determine the prevalence of type 2 diabetes and impaired glucose tolerance in subjects aged 40 years or more in Kashmir Valley, India. The study was carried out in two phases. In phase one, 6091 randomly selected subjects, 40 years or older, from all six districts of the valley were surveyed for prevalence of known diabetes mellitus. In phase two, 5083 subjects, 40 years or older, were screened with oral glucose tolerance test for prevalence of undiagnosed (asymptomatic) diabetes mellitus and impaired glucose tolerance. Abnormalities of carbohydrate intolerance were determined as recommended by WHO. Of 6091 subjects interviewed, 115 were known cases of diabetes mellitus with an overall prevalence of 1.89% (1.98% in males and 1.77% in females). Results of glucose tolerance test revealed that mean fasting as well as mean 2 h blood glucose was significantly more in females as compared to males (4.68+/-0.91 and 6.40+/-2.12 vs. 4.49+/-0.96 and 5.94+/-2.03 mmol/l, respectively, P < 0.0001). Of 5083 subjects who were subjected to glucose tolerance test (GTT), 627 (12.34%) had an abnormal test; with 411 (8.09%) having impaired glucose tolerance (IGT) and 216 (4.25%) having diabetes mellitus. The prevalence of IGT as well as of diabetes was significantly more in females as compared to males (P < 0.001). Subjects who had family history of diabetes had a significantly higher prevalence of abnormal GTT. Prevalence of known diabetes as well as that of abnormal GTT steadily increased with age, with a highest prevalence in the age group of > or = 70 years (P < 0.001). Obese subjects had a significantly higher basal as well as 2 h blood glucose in males as well as in females. Subjects with diabetes on GTT had a higher waist/hip ratio. Overall the prevalence of diabetes as well as IGT was significantly higher in the urban population. We conclude that 1.89% of the general population have known diabetes, 4.25% have undiagnosed diabetes and 8.09% have impaired glucose tolerance test; making the total load of abnormal glucose tolerance 14.23% in Kashmir Valley. In subjects greater than 40 years of age having a family history of diabetes, obesity, higher age (50 years or above), female sex, and urban origin have more chance (odds ratio: 4.65, 2.30, 1.87, 1.49 and 1.16, respectively) of developing abnormal glucose tolerance. PMID- 10670915 TI - No difference in serum sialic acid in type 2 diabetic patients from the United Arab Emirates with and without diabetic retinopathy. AB - Serum total sialic acid (TSA) has recently been shown to be related to diabetic retinopathy. However, there is some controversy as this may be true in European Type 2 diabetic patients but not South Asians. There are few data looking at serum TSA expression in Arab Type 2 diabetic patients from the United Arab Emirates (UAE) and we wished to test the hypothesis that there may be different serum TSA expression in Arab Type 2 diabetic patients as regard to retinopathy. Sixty-five Type 2 diabetic patients from the UAE were studied (19 male and 46 female, age 57.5+/-9.8 (45-74) years, duration of diabetes 9.4+/-5.7 (0-22) years. The serum TSA in 13 patients with diabetic retinopathy was 757+/-130 mg/l and 782+/-163 mg/l in those without retinopathy (NS). There was no significant correlation between serum TSA and patient age, serum fructosamine, diabetes duration, or blood pressure. As in South Asians serum TSA does not appear to be elevated in Type 2 diabetic patients from the United Arab Emirates with diabetic retinopathy. PMID- 10670916 TI - On the radioanalytical methods used to assay stainless-steel-encapsulated, ceramic-based 90Sr-90Y intravascular brachytherapy sources. AB - Very quantitative radiochemical procedures for the destructive assay of stainless steel-encapsulated, ceramic-based 90Sr-90Y intravascular brachytherapy sources (termed 'seeds') have been devised. These seeds, developed and provided by Bebig Isotopentechnic und Umweltdiagnostik (Berlin, Germany) in collaboration with the Novoste Corporation (Norcross, GA), are intended for use in the prophylactic treatment of restenosis following balloon angioplasty in heart-disease patients. The procedures were applied to the radionuclidic assay of both the bare-ceramic source materials (of proprietary composition) contained within the seeds and to the stainless-steel (SS) sealed sources. The approach consisted of extracting some arbitrary fraction of the 90Sr activity from the ceramic-like material and assaying the resulting solution by 4pi beta liquid scintillation (LS) spectrometry with 3H-standard efficiency tracing. The fraction of extracted activity was determined by ionization current measurements before and after the chemical extraction. All of the ionization current and LS-based activity determinations were made under the experimentally-verified conditional that 90Y was in radioactive equilibrium with 90Sr. For the assay of the SS-jacketed seeds, the encapsulation was initially dissolved and the resultant solution was also assayed by LS spectrometry to determine the amount of activity removed by the SS dissolution step. The developed protocol included provisions for accounting for all possible losses of 90Sr activity in the chemical and source-handling procedures, for the unrecovered activity in the extracted source material and for any residual activity in the solution-transfer and source-handling tools. These destructive assays were required for relating radiochromic-fill measurements of the absorbed dose spatial distributions for the seeds to theoretic dose modelling and for establishing calibration factors for subsequent non-destructive radionuclidic measurements on the seeds. PMID- 10670917 TI - Separation of 111Ag from neutron irradiated natural palladium using alumina as an adsorbent. AB - A simple method is presented for the separation of no-carrier-added 111Ag from neutron irradiated natural palladium. The method is based on sorption of 111Ag in 0.01 M HCl on alumina. Palladium is removed by washing with 0.1 M HCl and the 111Ag is eluted with 4 M HCl. The overall yields of 111Ag are better than 85% with < 1 microg/ml palladium as an impurity. The whole procedure from dissolving the target to the final 111Ag solution takes about 2 h. PMID- 10670918 TI - Evaluation of the contribution of smoking to total blood polonium-210 in Saudi population. AB - A preliminary study of 210Po concentrations in the blood of some smokers and nonsmokers is presented in order to evaluate the contribution of smoking to total blood 210Po in Saudi population. Blood samples were collected from 30 volunteers and analyzed by high resolution alpha-spectrometry using a radiochemical technique. The technique is based on the separation of polonium from other components of the sample by wet ashing with an HNO3/H2O2 oxidizing mixture and spontaneous deposition on a silver disc under the relevant conditions for alpha particle counting. The results indicated that a significant fraction (about 30%) of blood 210Po is related to smoking. PMID- 10670919 TI - Cleaning of liquid radioactive wastes using natural zeolites. AB - Natural zeolite, clinoptilolite, was used to eliminate liquid radioactive wastes (LRW) 137Cs and 90Sr. The influence of several factors (pH of solution, grain size of the zeolite, etc.) on the process effectivity was studied. It was shown that clinoptilolite is an effective filter of the nuclides above. PMID- 10670920 TI - Potential of rice husks for antimony removal. AB - The adsorption behavior of rice husks for antimony ions from aqueous solutions has been investigated as a function of appropriate electrolyte, equilibration time, hydrogen ions, amount of adsorbent, concentration of adsorbate, effect of diverse ions and temperature. The best conditions in which this material can be used as adsorbent have been explored. The radiotracer technique was employed to determine the distribution of antimony (122Sb) using a batch method. Maximum adsorption was observed at 0.01 mol L(-1) acid solutions (HNO3, HCl, H2SO4 and HClO4) using 1.0 g of adsorbent for 1.92 x 10(-5) mol L(-1) antimony concentration in 10 min equilibration time. Studies show that the adsorption decreases with the increase in the concentrations of all the acids. The adsorption data follow the Freundlich isotherm over the range of 1.92 x 10(-5) to 2.05 x 10(-4) mol L(-1) antimony concentration. The characteristic Freundlich constants i.e., 1/n = 0.82 +/- 0.05 and K = 4.61 +/- 0.07 m mol g(-1) have been computed for the sorption system. The uptake of antimony increases with the rise in temperature (299-323 K). Thermodynamic quantities i.e., deltaG0, deltaS0 and deltaH0 have also been calculated for the system. The sorption process was found to be endothermic. PMID- 10670921 TI - Investigation of defects in high-energy heavy ion implanted GaAs. AB - Undoped semi-insulating GaAs were implanted with 500 MeV Ne ions. Monte Carlo simulation revealed that the largest concentration of vacancies induced was around the end of the Ne ion range. Positron Annihilation measurement showed that after lower dose implantation, divacancies were formed, which coexisted with monovacancies. On increasing the dose, all the monovacancies changed to divacancies. The temperature dependence of positron lifetime suggested the existence of negatively charged antisites GaAs. Near infra-red spectra were also measured to study the implantation induced amorphous layers. PMID- 10670922 TI - Attenuation of fission neutrons by some hydrogeneous shield materials and the exponential dependence of the attenuated total neutron dose rate on the shield thickness. AB - This work deals with the attenuation of fission neutrons by some hydrogeneous shield materials. The attenuated fission neutrons are described by the energy groups (fast, epithermal and thermal). The exponential decrease in the fast flux is represented by the removal cross section concept. Each of the epithermal and thermal fluxes is expressed using the diffusion equation including a pair of arbitrary constants to be determined using the corresponding boundary conditions. The solution obtained for the required arbitrary constants is then approximated in a simplified form such that it may easily replace the corresponding exact solution. The attenuation values, by which the neutron dose rate distributions are exponentially decreased through certain thicknesses are also determined for the given materials. They are compared to the corresponding experimental and theoretical data. The results obtained for the total neutron dose rate distributions in terms of a suitable range of layer thicknesses are then used to determine--for each material--an average value for the total neutron dose rate representing the exponential decrease during passage through the considered range of layer thicknesses. PMID- 10670923 TI - Synthesis of [18F]fluoromethyl iodide, a synthetic precursor for fluoromethylation of radiopharmaceuticals. AB - [18F]fluoroiodomethane was labeled via nucleophilic substitution of diiodomethane with [18F]fluoride, and labeling conditions were optimized. The optimized labeling yield was 40 +/- 8% (decay-corrected). The synthesis and purification of [18F]fluoroiodomethane took 15 min. The reactions of [18F]fluoroiodomethane with amine, carboxylic acid, thiol and phenoxide groups produced fluoromethylated derivatives with various yields (12-95%). The results indicated that [18F]fluoroiodomethane is a valuable synthetic precursor for the introduction of an [18F]fluoromethyl group into radiopharmaceuticals. PMID- 10670924 TI - Preparation and biodistribution of rhenium-188 labeled albumin microspheres B 20: a promising new agent for radiotherapy. AB - Intra-arterial infusion of labeled particles is an effective method for endoradiotherapy of tumors. In this study, we radiolabeled biodegradable HSA microspheres (mean diameter = 25 microm) with the short-lived beta-emitter 188Re available from the aluminia-based 188W/188Re generator system. After 1 h 35-40% of the relative large amount of Sn(II) chloride required for effective reduction of Re(VII) for efficient attachment to the particles is precipitated as an amorphous coat of tin hydroxid colloid on the particle surface. The final 188Re bound to the particles was found to be stable in vitro. The radiolabelling yield was > 90%. The biological half-life was > 250 h and demonstrated sufficient in vivo stability after i.v. injection in Wistar rats. Because of the attractive properties of 188Re and the uniform particle size and stability, in vivo, this new agent is an attractive candidate for endoradiotherapy of tumors after selective catheterization. PMID- 10670925 TI - Syntheses and radiolabeling of cysteine-oximes and pharmacological behaviour of their 99mTc complexes. AB - Synthesis of two novel ligands using 2-oximino-butan-3-one and L-ethyl cysteinate is described. The synthetic procedure involved the formation of Schiffs base by the condensation of the amino group of L-ethyl cysteinate with the carbonyl group of 2-oximino-butan-3-one to provide the ligand I, N'(butan-2-enyl-3-oximino)ethyl cysteinate, followed by reduction of the Schiffs base with sodium borohydride to ligand II, N'(3-oximinobutyl)ethyl cysteinate. The ligands were characterised by NMR spectroscopy. Complexation studies with 99mTc were carried out using stannous tartrate as the reducing agent. The complexes were characterised by paper chromatography, thin layer chromatography and paper electrophoresis techniques. The complexes are formed in high yields when the reactions were carried out at pH 7-9. The 99mTc complex with ligand I is formed instantaneously while the 99mTc complex with ligand II is formed at a slower rate. The complexes were found to be neutral but the lipophilicity of the complex with ligand I was higher than that of the complex of ligand II. The stability of the complex with ligand I was relatively poor as compared to that of the complex with ligand II. Biodistribution studies of the 99mTc complexes of ligand I and II showed rapid blood clearance with hepatobiliary uptake. Renal excretion of the complex of ligand II was more than that observed for the complex of ligand I. The complexes did not show significant uptake in brain in spite of their favourable properties such as neutrality, lipophilicity and structural similarity with both ECD and HMPAO. PMID- 10670926 TI - In-target production of high specific radioactivity [15O]nitrous oxide by deuteron irradiation of nitrogen gas. AB - A simple and rapid production method for high specific radioactivity [15O]N2O has been developed based on the 14N(d,n)15O reaction on high-purity nitrogen gas in a flow-through target irradiated with a 0.5 microA beam of 7 MeV deuterons. The [15O]N2O formed during irradiation is selectively concentrated from the target effluent by adsorption on a zeolite during 150 s and subsequently released by rapid heating into a pulse with a full width at half maximum of 3.5 s. The radioactivity and specific radioactivity in the pulse amount to 4 MBq [15O]N2O and 4.5 x 10(13) Bq/mol respectively with a radiochemical purity >99.95%. A tenfold higher specific radioactivity may be feasible at larger beam currents. It was shown that stable N2O was also formed during irradiation. Based on responses to variations in various parameters during irradiation and on analyses performed on the products, an explanation is given on the mechanisms of in-target [15O]N2O and N2O formation, involving reaction of a particular excited state of O3 with N2. PMID- 10670927 TI - A new, convenient method for the preparation of 4-[18F]fluorobenzyl halides. AB - A convenient method suitable for automated preparation of 4-[18F]fluorobenzyl halides from no-carrier-added [18F]fluoride has been developed. 4 [18F]Fluorobenzaldehyde, synthesized from [18F]fluoride by aromatic nucleophilic substitution on 4-trimethylammoniumbenzaldehyde triflate, was first retained on a C18 cartridge and there efficiently reduced to 4-[18F]fluorobenzyl alcohol simply by flowing an aqueous solution of NaBH4. The conversion of 4-[18F]fluorobenzyl alcohol to 4-[18F]fluorobenzyl halide was investigated using PBr3, PI3, P2I4, Ph3PBr2 and Ph3PI2 in CH2Cl2. 4-[18F]Fluorobenzyl halides were purified by passing through a disposable silica cartridge. The conversion rapidly proceeded in radiochemical yields of nearly 90% at 40 degrees C with P2I4 and almost quantitatively at room temperature with Ph3PBr2. With this last reagent 4 [18F]fluorobenzyl bromide was obtained in overall radiochemical yields of 50-60% within 30 min from EOB. PMID- 10670928 TI - Investigations on thermal neutron reflection by activation method. AB - The prompt neutron detection and the foil activation methods were compared for the determination of the reflection cross-section of thermal neutrons and the hydrogen content of bulk samples. The advantages and limitations of the two methods are discussed. PMID- 10670929 TI - Effect of lithium co-dopant on the thermoluminescence response of some phosphors. AB - The effect of lithium co-dopant on the thermoluminescence of some TL phosphors has been studied. These results show that lithium co-dopant when present in CaSO4:Dy, CaSO4,Tm, CaF2:Mn, MgB4O7:Dy, MgB4O7:Tm, MgB4O7:Tb, MgB4O7:Mn, CaB4O7:Dy and CaB4O7:Tm thermoluminescent phosphors induces higher luminescence efficiency either due to better incorporation of activator ions or due to improvement in energy transfer processes. Co-doping with Li of the above given phosphors resulted in increasing the TL sensitivity from 1.2 to 3.4 times, when compared with that of the TL phosphors without lithium co-dopant. The effect of co-doping with lithium impurities on the thermoluminescence of various TL phosphors is also studied in dependence of Li concentration, of the chemical form of added lithium compounds and of the method of addition of Li co-dopant during the TL phosphor preparation. It is observed that the addition of Li co-dopant induces in some TL phosphors enhancement of the lower temperature peak which is correlated with the increase of TL sensitivity. The high TL output of these TL phosphors could be used in dosimetric practice for special short-term measurements. PMID- 10670930 TI - An iterative approach for TRIGA fuel burn-up determination using nondestructive gamma-ray spectrometry. AB - The purpose of this work is to establish a method for evaluating the burn-up values of the rod-type TRIGA spent fuel by using gamma-ray spectrometry of the short-lived fission products 97Zr/97Nb, 132I, and 140La. Fuel irradiation history is not needed in this method. Short-lived fission-product activities were established by reirradiating the spent fuels in a nuclear reactor. Based on the measured activities, 235U burn-up values can be deduced by iterative calculations. The complication caused by 239Pu production and fission is also discussed in detail. The burn-up values obtained by this method are in good agreement with those deduced from the conventional method based on long-lived fission products 137Cs, 134Cs/137Cs ratio and 106Ru/137Cs ratio. PMID- 10670931 TI - Assessing the information content of phosphor produced medical images: application to Zn2SiO4:Mn phosphor. AB - In this study a method to assess the information content of medical images produced by phosphors is described. The optical signal emitted by the phosphor after X-ray excitation, the detective quantum efficiency (DQE), expressing the signal-to-noise ratio (SNR) transfer efficiency, and the information capacity were experimentally determined. The method was based on light flux and modulation transfer function (MTF) measurements and was used to assess the imaging performance of the Zn2SiO4:Mn phosphor. The latter was employed in the form of laboratory prepared phosphor layers (test screens). Results showed that high values for optical signal emission and DQE were obtained for medium thickness phosphor layers (56 and 89 mg/cm2) at 20 kVp X-ray tube voltage. The information capacity was found to decrease continuously with phosphor coating weight. PMID- 10670932 TI - Borehole parametric study for neutron induced capture gamma-ray spectrometry using the MCNP code. AB - The MCNP Monte Carlo code has been used to simulate neutron transport from an Am Be source into a granite formation surrounding a borehole. The effects of the moisture and the neutron poison on the thermal neutron flux distribution and the capture by the absorbing elements has been calculated. Thermal and nonthermal captures for certain absorbers having resonance structures in the epithermal and fast energy regions such as W and Si were performed. It is shown that for those absorbers having large resonances in the epithermal regions when they are present in dry formation or when accompanied by neutron poisons the resonance captures may be significant compared to the thermal captures. PMID- 10670933 TI - Indoor radon/thoron survey report from Hamirpur and Una districts, Himachal Pradesh, India. AB - A survey of indoor radon and thoron levels has been carried out in a number of villages in the vicinity of uranium bearing sites in the Hamirpur and Una districts of Himachal Pradesh (H.P.), India. Levels were analysed with reference to the nature of building material, soil type and different seasons of the year. The one year average for radon concentration was found to vary from a minimum of 19.7 to a maximum of 146.3 Bq/m3 while the minimum and maximum thoron concentrations were 9.1 and 70.7 Bq/m3, respectively. The dose rate varied from 0.1 to 8.67 microSv/h. These are discussed in the light of ICRP recommendations. PMID- 10670934 TI - Instrumental neutron activation analysis of rocks from Cayajabos petroleum ore. AB - The relative INAA method under statistical control was used to measure the concentrations of Sb, La, Eu, Co, Rb, Tb, Cs, Hf, Th, Cr, Lu, Yb, Ce, Sm, Gd, Nd. Rare earth element (REE) Chondrite-normalized patterns are presented. The La(cond)/Lu(cond) normalized concentration ratios obtained confirm the carbonaceous character of the rocks. PMID- 10670935 TI - Comparison between different types of glass and aluminum as containers for irradiation samples by neutron activation analysis. AB - Three different types of glass and four different kinds of aluminum sheet have been analyzed using neutron activation analysis. The irradiation facilities of the first Egyptian research reactor (ET-RR-1) and a hyper-pure germanium (HPGe) detection system were used for the analysis. Among the 34 identified elements, the isotopes 60Co, 65Zn, 110mAg, 123mTe, 134Cs, 152Eu and 182Ta are of special significance because of their long half-lives, providing a background interference for analyzed samples. A comparison between the different types of containers was made to select the preferred one for sample irradiation. PMID- 10670936 TI - Studies on microencapsulation of 5-fluorouracil with poly(ortho ester) polymers. AB - Microencapsulation of 5-fluorouracil was successfully accomplished with poly(ortho ester) polymers by the emulsification-solvent evaporation method. While actual drug loading increased with increasing drug load (5-15% w/w), the entrapment efficiency remained essentially unaffected, under a given set of experimental conditions. Incorporation of sorbitan sesquioleate enhanced entrapment efficiency, decreased the volume-surface mean diameter of the poly(ortho ester) microspheres and provided controlled release of 5-fluorouracil. The volume of the aqueous phase was more important than the concentration of polyvinyl alcohol in it. The entrapment efficiency improved from 13 to 33% when the volume of the aqueous phase was increased from 20 to 80 ml. The volume of organic phase (methylene chloride) and the concentration of polymer in it played an important role. The use of smaller volumes of more concentrated polymer solution enhanced actual drug loading, entrapment efficiency and produced larger microspheres. The release studies conducted in 0.01 M phosphate buffer at 37+/ 1.0 degrees C demonstrated that the release of 5-FU from the microspheres prepared with sorbitan sequioleate was nearly independent of the initial drug load with a mean zero-order rate constant of 0.0063% per hour. The data suggested that drug release was largely a diffusional process with contributions from dissolution and polymer degradation. PMID- 10670937 TI - In vitro delivery of a sparingly water soluble compound from PLA50 microparticles. AB - The administration of a sparingly soluble drug is always problematic, especially when the drug has to be released from the degradable matrix of a polymeric drug delivery system. Attempts were made to achieve the complete release of 1-[2 (fluorobenzoyl) aminoethyl]-4-(7-methoxynaphtyl)piperazine (FAMP), a potential anxiolytic and antidepressor hydrophobic compound, from racemic poly(lactic acid) (PLA50)-based microparticles, 100% release was required at a low rate in order to allow monthly repeated S.C. or I.M. injections of this potent compound. FAMP polymer combinations were made in the form of microspheres by the solvent evaporation technique. Release profiles were investigated under dynamic conditions by using a constant flow rate of pH 7.4 0.15 M phosphate buffer, used as a model of body fluids. Under these conditions, none of the microsphere compositions led to total release within a month, even when hydrophilic excipients, namely fructose and PEG were added. PLA50-FAMP microparticles with compositions and sizes similar to those of the microspheres, were then made by direct blending in dichloromethane, evaporation of the solvent, grinding and sieving. These formulations also failed in providing total drug release within 30 days, even at a high drug load. FAMP/PLA50/water-soluble additive, ternary grounded particles were finally prepared with fructose, PLA50 oligomers or poly(ethylene glycol) (PEG) as the additive. Only PLA50 grounded particles with percolating FAMP-PEG microdomains allowed 100% release of FAMP over a 30 day period, at a quasi constant rate which depended primarily on solubility and channelling provided the flow was slow enough. Data are discussed in terms of the accessibility of the entrapped drug to the aqueous medium. PMID- 10670938 TI - Preparation and some properties of water-insoluble, whey protein-based microcapsules. AB - A method, consisting of double emulsification and chemical cross-linking with glutaraldehyde was used to prepare whey protein-based microcapsules containing anhydrous milk fat as a model core. Effects of emulsion composition and pH on core retention, microstructure, and water-solubility of microcapsules were investigated. In all cases, core retention higher than 88% was accomplished and, in most cases, was not significantly affected by emulsion composition. In all cases, spherical microcapsules, 10-80 microm in diameter, were obtained. Outer topography and the inner structure of microcapsules were significantly affected by the pH of the emulsion. In all cases, microcapsules were practically water insoluble. Microcapsules similar to the developed prototype may be suitable for controlled core release in application fields where chemical cross-linking is acceptable. PMID- 10670939 TI - Poly(vinylbenzyl chloride) microsphere synthesis and their chemical modifications. AB - Vinylbenzyl chloride (VBC) was dispersion polymerized to give monodisperse microspheres in the presence of poly(vinylpyrrolidone) (PVP) as a steric stabilizer. The effect of PVP concentration on the size and on microsphere stability during the polymerization process was investigated. Microsphere size was examined when co-stabilizer molecules were employed with PVP during the polymerization reaction. The built-in reactive chloromethyl groups of the microspheres were the sites of the nucleophilic reaction of two amino-group model molecules, glucosamine (G), a hexosamine implicated in processes of molecular recognition, and also bovine serum albumin (BSA). Elemental analyses and Fourier transform infrared spectroscopy (FTIR) spectra showed that poly(vinylpyrrolidone) was associated with the microsphere network. Elemental analyses, attenuated total reflection infrared spectroscopy (ATR-FTIR), and zeta potential measurements confirmed G and BSA links at the microsphere surface. PMID- 10670940 TI - Studies on the in vitro release characteristics of ibuprofen-loaded polystyrene microparticles. AB - Ibuprofen-loaded polystyrene microparticles were prepared by the emulsion-solvent evaporation process from an aqueous system. The effects of different parameters on the drug content and on the release of the drug from the microparticles were investigated. The drug content, in all the formulations, was less than the theoretical drug loading. The lower drug content was due to drug partitioning to the external aqueous phase during formulation. Statistical analysis revealed that the variation in the concentrations of the emulsion stabilizer and the organic disperse phase volume did not significantly alter the release of the drug. Although an increase in drug loading increased drug release from the microparticles, a biphasic linear relationship was observed between the time required for 50% drug release and the drug loading. The effect of size of the microparticles on drug release was more important for the low drug-loaded microparticles than that for the high drug-loaded microparticles. Such release behaviour from the microparticles was explained on the basis of the morphological structure of the microparticles. PMID- 10670941 TI - Long-term stability of PBCA nanoparticle suspensions. AB - In this study, the stability of poly(butyl cyanoacrylate) (PBCA) nanoparticle suspensions was examined for up to 1 year by measuring the nanoparticle sizes. The nanoparticles were prepared with different stabilizers (dextran 70.000, poloxamer 188, or polysorbate 85), and the particle size was determined before and after purification by centrifugation and after dilution with different solutions (0.1 N HCl, 0.01 N HCl, H2O, and PBS). The most constant sizes were with the untreated acidic nanoparticle suspensions. In all other cases, agglomeration of the particles occurred: the extent of this agglomeration and the time at which the agglomeration occurred depended on the experimental conditions. Nanoparticle polymer degradation, as indicated by size decrease, was not observed. Thus, PBCA nanoparticles can be stored as suspensions, making the lyophilization and the sometimes problematic resuspension by ultrasonication, unnecessary, which is advantageous for clinical applications. PMID- 10670942 TI - The use of additives to modulate the release of a sparingly water soluble drug entrapped in PLA50 microparticles. AB - One of the major problems raised by the microencapsulation of drugs which are sparingly soluble in water is the difficulty to achieve a controlled and total release of the drug. It was previously shown that the microencapsulation of a model water insoluble drug, namely 1-[2-(4-fluorobenzoyl)aminoethyl]-4-(7 methoxynaphthyl) piperazine hydrochloride (FAMP) with a hydrophilic additive like low molar mass poly(ethylene glycol)s (PEG) can fulfil these requirements, provided all the drug + additive matter is in contact with the surrounding liquid medium via open pores and percolating channels. In this paper, PEG was replaced by other additives, selected because of their potential ability to increase the solubility of FAMP in pH = 7.4 isosomolar phosphate buffer (PBS). The idea was that increasing the solubility locally in microparticles could allow the drug to be released, despite its poor solubility in aqueous media like body fluids, and be absorbed before recrystallization. The solubility in PBS of FAMP mixed with additive, in the form of solid dispersions, was determined for various additives, namely citric acid, dimyristoyl DL-alpha-phosphatidyl choline (DMPC), poloxamer copolymers of different compositions and poly(dodecyl L-lysine citramidate) (PLCAC12(100)), an aggregate-forming hydrophilic polyelectrolyte containing 100%, hydrophobizing ester groups which can accommodate lipophilic compounds in hydrophobic pockets present in the aggregates. PEG was taken as a reference. It was found that DMPC, some poloxamers and the hydrophobized polyelectrolyte do increase the solubility of FAMP in PBS. Investigation was made of the release of FAMP from ground microparticles, whose loads were composed of FAMP combined with these solubilization-promoting additives. It was found that the release rate of FAMP from such systems can be increased and modulated to achieve an in vitro sustained release over a 20-30 day period and secure exhaustion of the particles at the end of this period. PMID- 10670943 TI - The use of additives to modulate the release of a sparingly water soluble drug entrapped in PLA50 microparticles: in vivo investigation. AB - Sustained and total release of the sparingly water soluble compound, namely 1-[2 (4-fluorobenzoyl)aminoethyl]-4-(7-methoxynaphthyl) piperazine hydrochloride (FAM), from poly (DL-lactic acid) (PLA50) microparticles was previously shown to be feasible if the particles are obtained by grinding a solid mixture composed of the polymer and a percolating array of the compound mixed with an additive. Such microparticles, where the additive was poly (ethylene glycol) (PEG), dimyristoylphosphatidylcholine (DMPC), or Poloxamer 6800, were administrated subcutaneously to rats either as depot or using a liquid vehicle. The variations of the plasma concentration vs time determined by high pressure liquid chromatography and fluorometric detection, were plotted for the various microparticle systems, blood being taken twice from each animal and each measurement being triplicated. Data were analysed by non-compartmental analysis, in order to evaluate the elimination constant, the half-life, the area under the curve and the bioavailability for each system. Kinetics experiments were performed over 24h and also for 7 days. It was found that, for the selected formulations, the release of the sparingly water soluble compound depends on the dissolution rate in vivo and on the physicochemical characteristics of the additive, including solubility and micelle formation. Data correlated well with the results of previous in vitro investigation. PMID- 10670944 TI - Preparation of microcapsules for skin allergy testing by the solvent evaporation process. AB - Polyurethanes and polyvinyl alcohol modified by stearyl isocyanate are used as a matrix for microparticles made by a solvent evaporation process to encapsulate allergenic molecules, with petrolatum used as a neutral vehicle. The encapsulation yields, depending on the agent to be encapsulated, vary from 22 to 45%. PMID- 10670945 TI - Pea chloroplast carnitine acetyltransferase. AB - The purpose of this study was to resolve the controversy as to whether or not chloroplasts possess the enzyme carnitine acetyltransferase (CAT) and whether the activity of this enzyme is sufficient to support previously reported rates of fatty acid synthesis from acetylcarnitine. CAT catalyses the freely reversible reaction: carnitine + short-chain acylCoA <--> short-chain acylcarnitine + CoASH. CAT activity was detected in thc chloroplasts of Pisum sativum L. With membrane impermeable acetyl CoA as a substrate. activity was only detected in ruptured chloroplasts and not with intact chloroplasts, indicating that the enzyme was located on the stromal side of the envelope. In crude preparations, CAT could only be detected using a sensitive radioenzymatic assay due to competing reactions from other enzymes using acetyl CoA and large amounts of ultraviolet absorbing materials. After partial purification of the enzyme, CAT was detected in both the forward and reverse directions using spectrophotometric assays. Rates of 100 nmol of product formed per minute per milligram of protein were obtained, which is sufficient to support reported fatty acid synthesis rates from acetylcarnitine. Chloroplastic CAT showed optimal activity at pH 8.5 and had a high substrate specificity, handling C2-C4 acyl CoAs only. We believe that CAT has been satisfactorily demonstrated in pea chloroplasts. PMID- 10670946 TI - Cirri of the stalked crinoid Metacrinus rotundus: neural elements and the effect of cholinergic agonists on mechanical properties. AB - Sea lilies are enigmatic animals due to their scarcity and their biology is comparatively neglected. Cirri, arranged in whorls of five along the sea lily stalk, anchor and support the animal. They consist of ossicles interconnected by collagenous ligaments and by a central canal. Cirri have a well-developed nervous system but lack muscular cells. A light and electron microscopic study was performed to clarify the morphology of the nervous system of the cirri. Two cellular networks were found, one of neuron-like cells and one of cells filled with bullet-shaped organelles. Both networks ramify throughout the cirral ossicles up to the interossicle ligaments. Mechanical tests were performed to analyse the influence of cholinergic agonists on the mechanical properties of these ligaments. In the tests, the cirral ligaments softened after the application of acetylcholine, muscarinic agonists and nicotinic agonists. The reaction time to muscarinic agonists was much slower than to acetylcholine and nicotinic agonists. At low concentrations, muscarinic agonists caused active development of force. No reaction to stimuli was observed in anaesthetized cirri. The data clearly establish the existence of catch connective tissue which can change its mechanical properties under nervous control mediated via nerves with cholinergic receptors. The possible sources of the observed force production are discussed and it is concluded that active contraction of collagenous ligaments causes movement of cirri. PMID- 10670947 TI - Coalition formation in animals and the nature of winner and loser effects. AB - Coalition formation has been documented in a diverse array of taxa, yet there has been little formal analysis of polyadic interactions such as coalitions. Here, we develop an optimality model which examines the role of winner and loser effects in shaping coalition formation. We demonstrate that the predicted patterns of alliances are strongly dependent on the way in which winner and loser effects change with contestant strength. When winner and loser effects decrease with the resource-holding power (RHP) of the combatants, coalitions will be favoured between the strongest members of a group, but not between the weakest. If, in contrast, winner and loser effects increase with RHP, exactly the opposite predictions emerge. All other things being equal, intervention is more likely to prove worthwhile when the beneficiary of the aid is weaker (and its opponent is stronger), because the beneficiary is then less likely to win without help. Consequently, intervention is more probable when the impact of victory on the subsequent performance of a combatant increases with that individual's strength because this selects for intervention in favour of weaker combatants. The published literature on hierarchy formation does not reveal how winner and loser effects actually change with contestant strength and we therefore hope that our model will spur others to collect such data; in this light we suggest an experiment which will help to elucidate the nature of winner and loser effects and their impact on coalition formation in animals. PMID- 10670948 TI - Assessment of the prevalence of vCJD through testing tonsils and appendices for abnormal prion protein. AB - The objective of this study was to determine the age group or groups which will provide the most information on the potential size of the vCJD epidemic in Great Britain via the sampling of tonsil and appendix material to detect the presence of abnormal prion protein (PrP(Sc)). A subsidiary aim was to determine the degree to which such an anonymous age-stratified testing programme will reduce current uncertainties in the size of the epidemic in future years. A cohort- and time stratified model was used to generate epidemic scenarios consistent with the observed vCJD case incidence. These scenarios, together with data on the age distribution of tonsillectomies and appendectomies, were used to evaluate the optimal age group and calendar time for undertaking testing and to calculate the range of epidemic sizes consistent with different outcomes. The analyses suggested that the optimal five-year age group to test is 25-29 years, although a random sample of appendix tissue from all age groups is nearly as informative. A random sample of tonsil tissue from all age groups is less informative, but the information content is improved if sampling is restricted to tissues removed from those over ten years of age. Based on the assumption that the test is able to detect infection in the last 75% of the incubation period, zero detected infections in an initial random sample of 1000 tissues would suggest that the epidemic will be less than 870,000 cases. If infections are detected, then the model prediction suggests that both relatively small epidemics (800+ cases if one is detected or 8300+ if two are detected) and larger epidemics (21,000+ cases if three or more are detected) are possible. It was concluded that testing will be most informative if undertaken using appendix tissues or tonsil tissues removed from those over ten years of age. Large epidemics can only be excluded if a small number of infections are detected and the test is able to detect infection early in the incubation period. PMID- 10670949 TI - Longevity and the costs of reproduction in a historical human population. AB - It has been argued that the priority that natural selection places on reproduction negatively affects other processes such as longevity and the problem posed by this trade-off underlies the disposable soma theory for the evolution of human ageing. Here we examine the relationship between reproduction and longevity in a historical human population (the Krummhorn, north-west Germany 1720-1870). In our initial analyses, we found no support for the hypothesized negative effects of reproduction on longevity: married women who remained childless lived no longer than women who reproduced and women who had few children lived no longer than women who had many children. However, more detailed analyses in relation to socio-economic class revealed that the extent to which reproduction has an effect on longevity is a function of the level of economic deprivation. We found that, when possible sources of confound were controlled for (e.g. duration of marriage and amount of time spent in fecund marriage), there is an increasingly strong relationship between longevity and reproduction with increasing poverty. PMID- 10670950 TI - Resource allocation between reproductive phases: the importance of thermal conditions in determining the cost of incubation. AB - Changes in the resources allocated to particular stages of reproduction are expected to influence allocation to, and performance in, subsequent reproductive stages. Experimental manipulation of individual investment patterns provides important evidence that such physiological trade-offs occur, and can highlight the key environmental variables that influence reproductive costs. By temporarily altering the thermal properties of starling nests, we reduced the energetic demand of first-clutch incubation, and examined the effect of this manipulation on performance during the same and the subsequent reproductive attempts. Compared with controls, starlings investing less in incubation were more successful in fledging young, and were more likely to hatch all their eggs if a subsequent reproductive attempt was made. Our results show that incubation demands can limit reproductive success, and that resources saved during incubation can be reallocated to later stages of the same reproductive attempt and to future reproductive attempts. This study also shows that small changes in thermal environment can affect breeding success by altering the energetic demands imposed on incubating parents, independently of the effect of temperature on other environmental variables such as food supply. PMID- 10670951 TI - Stable isotopes examined across a migratory divide in Scandinavian willow warblers (Phylloscopus trochilus trochilus and Phylloscopus trochilus acredula) reflect their African winter quarters. AB - The C and N isotopes of feathers from two subspecies of willow warblers Phylloscopus trochilus trochilus and Phylloscopus trochilus acredula) are isotopically distinct. Our analysis of 138 adult males from 14 sites distributed across Sweden shows that the mean delta15N and delta13C values of subspecies acredula (from latitudes above 63 degrees N) were significantly higher than the mean delta15N and delta13C values of subspecies trochilus (from latitudes below 61 degrees N). The analysed willow warbler feathers had been moulted in the winter quarters and the observed isotopic signatures should thus reflect the isotopic pattern of food assimilated in Africa. The isotopic data observed in Sweden match the cline in morphology, both showing abrupt changes around 62 degrees N. This result agrees with data from ringing recoveries indicating that the two subspecies occupy geographically and isotopically distinct wintering grounds in Africa. Our isotopic data suggest that analysis of stable isotopes of C and N is a promising method to track wintering quarters of European birds that migrate to Africa. PMID- 10670952 TI - Robustness of reserve selection procedures under temporal species turnover. AB - Complementarity-based algorithms for the selection of reserve networks emphasize the need to represent biodiversity features efficiently, but this may not be sufficient to maintain those features in the long term. Here, we use data from the Common Birds Census in Britain as an exemplar data set to determine guidelines for the selection of reserve networks which are more robust to temporal turnover in features. The extinction patterns found over the 1981-1991 interval suggest that two such guidelines are to represent species in the best sites where they occur (higher local abundance) and to give priority to the rarer species. We tested five reserve selection strategies, one which finds the minimum representation set and others which incorporate the first or both guidelines proposed. Strategies were tested in terms of their efficiency (inversely related to the total area selected) and effectiveness (inversely related to the percentage of species lost) using data on eight pairs of ten-year intervals. The minimum set strategy was always the most efficient, but suffered higher species loss than the others, suggesting that there is a trade-off between efficiency and effectiveness. A desirable compromise can be achieved by embedding the concerns about the long-term maintenance of the biodiversity features of interest in the complementarity-based algorithms. PMID- 10670953 TI - Better red than dead: carotenoid-based mouth coloration reveals infection in barn swallow nestlings. AB - Nestling birds solicit food from their parents by displaying their open brightly coloured gapes. Carotenoids affect gape colour, but also play a central role in immunostimulation. Therefore, we hypothesize that, by differentially allocating resources to nestlings with more brightly coloured gapes, parents favour healthy offspring which are able to allocate carotenoids to gape coloration without compromising their immune defence. We demonstrated that, in the barn swallow Hirundo rustica, (i) parents differentially allocate food to nestlings with an experimentally brighter red gape, (ii) nestlings challenged with a novel antigen (sheep red blood cells, SRBCs) have less bright gape colour than their control siblings, (iii) nestlings challenged with SRBCs but also provided with the principal circulating carotenoid (lutein) have more brightly coloured red gapes than their challenged but unsupplemented siblings and (iv) the gape colour of nestlings challenged with SRBCs and provisioned with lutein exceeds that of siblings that were unchallenged. This suggests that parents may favour nestlings with superior health by preferentially feeding offspring with the brightest gapes. PMID- 10670954 TI - Density-dependent aposematism in the desert locust. AB - The ecological processes underlying locust swarm formation are poorly understood. Locust species exhibit phenotypic plasticity in numerous morphological, physiological and behavioural traits as their population density increases. These density-dependent changes are commonly assumed to be adaptations for migration under heterogeneous environmental conditions. Here we demonstrate that density dependent nymphal colour change in the desert locust Schistocerca gregaria (Orthoptera: Acrididae) results in warning coloration (aposematism) when the population density increases and locusts consume native, toxic host plants. Fringe-toed lizards (Acanthodactylus dumerili (Lacertidae)) developed aversions to high-density-reared (gregarious-phase) locusts fed Hyoscyamus muticus (Solanaceae). Lizards associated both olfactory and visual cues with locust unpalatability, but only gregarious-phase coloration was an effective visual warning signal. The lizards did not associate low rearing density coloration (solitarious phase) with locust toxicity. Predator learning of density-dependent warning coloration results in a marked decrease in predation on locusts and may directly contribute to outbreaks of this notorious pest. PMID- 10670955 TI - Sex-ratio-distorting Wolbachia causes sex-role reversal in its butterfly host. AB - Sex-role-reversed mating systems in which females compete for males and males may be choosy are usually associated with males investing more than females in offspring. We report that sex-role reversal may also be caused by selfish genetic elements which distort the sex ratio towards females. Some populations of the butterflies Acraea encedon and Acraea encedana are extremely female biased because over 90% of females are infected with a Wolbachia bacterium that is maternally inherited and kills male embryos. Many females in these populations are virgins suggesting that their reproductive success may be limited by access to males. These females form lekking swarms at landmarks in which females exhibit behaviours which we interpret as functioning to solicit matings from males. The hypothesis that female A. encedon swarm in order to mate is supported by the finding that, in release recapture experiments, mated females tend to leave the swarm while unmated females remained. This behaviour is a sex-role-reversed form of a common mating system in insects in which males form lekking swarms at landmarks and compete for females. Female lekking swarms are absent from less female-biased populations and here the butterflies are instead associated with resources in the form of the larval food plant. PMID- 10670956 TI - Genetic support for the evolutionary theory of reproductive transactions in social wasps. AB - Recent evolutionary models of reproductive partitioning within animal societies (known as 'optimal skew', 'concessions' or 'transactional' models) predict that a dominant individual will often yield some fraction of the group's reproduction to a subordinate as an incentive to stay in the group and help rear the dominant's offspring. These models quantitatively predict how the magnitude of the subordinate's 'staying incentive' will vary with the genetic relatedness between dominant and subordinate, the overall expected group output and the subordinate's expected output if it breeds solitarily. We report that these predictions accord remarkably well with the observed reproductive partitioning between conesting dominant and subordinate queens in the social paper wasp Polistes fuscatus. In particular, the theory correctly predicts that (i) the dominant's share of reproduction, i.e. the skew, increases as the colony cycle progresses and (ii) the skew is positively associated both with the colony's productivity and with the relatedness between dominant and subordinate. Moreover, aggression between foundresses positively correlated with the skew, as predicted by transactional but not alternative tug-of-war models of societal evolution. Thus, our results provide the strongest (quantitative support yet for a unifying model of social evolution. PMID- 10670957 TI - A simple method of removing the effect of a bottleneck and unequal population sizes on pairwise genetic distances. AB - In this paper, we derive the expectation of two popular genetic distances under a model of pure population fission allowing for unequal population sizes. Under the model, we show that conventional genetic distances are not proportional to the divergence time and generally overestimate it due to unequal genetic drift and to a bottleneck effect at the divergence time. This bias cannot be totally removed even if the present population sizes are known. Instead, we present a method to estimate the divergence times between populations which is based on the average number of nucleotide differences within and between populations. The method simultaneously estimates the divergence time, the ancestral population size and the relative sizes of the derived populations. A simulation study revealed that this method is essentially unbiased and that it leads to better estimates than traditional approaches for a very wide range of parameter values. Simulations also indicated that moderate population growth after divergence has little effect on the estimates of all three estimated parameters. An application of our method to a comparison of humans and chimpanzee mitochondrial DNA diversity revealed that common chimpanzees have a significantly larger female population size than humans. PMID- 10670958 TI - Glucosinolate genetics and the attraction of the aphid parasitoid Diaeretiella rapae to Brassica. AB - The control of insect pests using parasitoids and carnivores has been successfully applied in protected cropping systems, orchards and forestry. Their success in annual field crops has been more limited due largely to the difficulties of attracting and maintaining a sufficient density of parasitoids in the crop before the levls of the insect herbivores become economically damaging. Parasitoids are known to be attracted to host-plant volatiles; thus, manipulating the host-plant chemistry may provide a means of enhancing the attraction of parasitoids to their prey. In this study we describe the differential attraction of the braconid wasp Diaeretiella rapae to two near-isogenic lines of Brassica oleracea which differ in a gene which alters the chemical structure of the isothiocyanates which are emitted following tissue damage. We demonstrate that, by enhancing the production of but-3-enyl isothiocyanate in B. oleracea and Brassica napus (oilseed rape), we can increase the attraction of D. rapae to these plants under standard field conditions. PMID- 10670959 TI - Loss of genetic diversity in the endemic Hector's dolphin due to fisheries related mortality. AB - The endemic New Zealand Hector's dolphin is considered the rarest species of marine dolphin with a total abundance of less than 4000. The species is listed as vulnerable because of fisheries-related mortality due to entanglement in set nets. The vulnerability of this species is further increased by its fidelity to local natal ranges and the genetic isolation of regional populations. Here we present evidence, based on 108 contemporary samples and 55 historical samples dating back to 1870, of a significant loss of mitochondrial DNA (mtDNA) diversity in two regional populations of Hector's dolphin. The haplotype diversity (h) was calculated from sequences of a 206 bp fragment in the mtDNA control region, designed to identify 13 out of the 14 known maternal lineages. Over the last 20 years, the North Island population has been reduced from at least three lineages (h = 0.41) to a single lineage (h = 0; p < 0.05). Given its small size, reproductive isolation and reduced genetic diversity, this population is likely to become extinct. The diversity of the East Coast South Island population has declined significantly from h = 0.65 to h = 0.35 (p < 0.05). Based on trend analysis of the mtDNA diversity, we predict that the East Coast population will lose all mtDNA diversity within the next 20 years. This time-series of reduction in genetic variation provides independent evidence of the severity of population decline and habitat contraction resulting from fisheries and perhaps other human activities. PMID- 10670960 TI - Environmental health research: setting an agenda by spinning our wheels or climbing the mountain? AB - This paper examines the nature and characteristics of research in environmental health, viewed as the effects of the environment on human health. It is argued that most of this work has been predicated on an epidemiological approach which has yielded significant (if sometimes equivocal) findings about exposure-outcome relationships. This discussion, however, concentrates on the limited and somewhat partial view of theory implied in this perspective. It advocates instead a broad based approach to theory as the basis for understanding significant portions of the social world. It posits, as illustrations, several social theories and with examples tries to show how environmental health research might be different. PMID- 10670961 TI - Managing risk in the city: the role of welfare professionals in managing risks arising from vulnerable individuals in cities. AB - Modern cities depend on individualism and the process of contracting. Contracts between individuals contribute to stability and order in cities. However, this is challenged by risks and uncertainties especially those relating to vulnerable individuals who are unable or unwilling to enter into contractual relations. This paper focuses on the role of caring professions and the impact of new strategies for managing risks related to vulnerable adults and children, especially the shift from managing risk in institutions to managing risk in the community. This paper is based on research funded through the ESRC Risk and Human Behaviour Programme. PMID- 10670962 TI - Physician retention in rural communities: the perspective of experiential place integration. AB - Rural communities across the United States continue to struggle in their attempts to recruit and retain physicians. In response to this ongoing problem, health services researchers have focused on the recruitment of physicians. More recently, however, researchers have recognized the importance of physician retention. Nonetheless, there is a scarcity of theories to explain the process of rural physician retention. This paper provides an overview of retention in health services research. It then proposes a theoretical perspective on retention called "experiential place integration." The paper subsequently presents an in-depth qualitative study of rural physician integration in Kentucky from which a practical framework of physician integration is constructed. The framework represents integration as an active developmental process based on the enhancement of security, freedom, identity and meaning in place. PMID- 10670963 TI - Image-diagnosis: visualization of community health levels and the impacts of determinants thereof. AB - The objective of this study is to develop a tool that visualizes health-level of communities and the determinants thereof. We developed Image-Diagnosis, which consists of mapping and star-plotting procedures. This paper presents this method by applying the statistics in Tokyo. The mapping shows the spatial variance of health-level and health-determining factors; star-plotting depicts health-level, as the circle, and health-determining factors, as the arms, in a star-shaped diagram. The animation of star-plots presents variations in health levels resulting from the specified changes in health-determining factors according to the models. Potential contribution to the decision making in urban health policy was discussed. PMID- 10670964 TI - A turning tide? Reflections on ideology and health service restructuring in New Zealand. PMID- 10670965 TI - Social aspects of AIDS-related stigma in rural Uganda. AB - In the process of collecting sexual behaviour data through in-depth interviews, 24 respondents offered information on stigma related to HIV-1 infection. Observations of social relations in public places and families of infected individuals were made. The findings suggest that although HIV/AIDS-related stigma has had adverse effects on treatment seeking behaviour of PWAs and coping mechanisms of their families, a more tolerant attitude is starting to emerge in this area. Probably, due to improvements in counselling services and home care schemes for those with AIDS. This supports the call for increased investments in counselling and community development aimed at caring for people with AIDS (PWAs). PMID- 10670966 TI - Ageing and aged care in the People's Republic of China: national and local issues and perspectives. AB - China's population is rapidly ageing at a time when former socialist collective provision and provision by the state in all sectors, especially in social welfare, is being radically reduced because of economic reform and financial stringency. The traditional Chinese approach to family care for elderly members is being encouraged but may be difficult because of smaller family sizes and the disruption of migration. This paper discusses some urban responses to pressures for change in care of elderly people, drawing on the example of Guangzhou (Canton) in southern China, which typifies many of the problems of caring for elderly people in times of social and economic change. It notes the development of homes and facilities for elderly people and the emergence of some prestige homes, often occupied by the better off, which have received both local and international investment. By contrast, the bulk of elderly people will not be adequately provided for by a declining public/collective sector. The dilemmas faced by the Chinese authorities attempting to stimulate local provision for all elderly people are identified. PMID- 10670967 TI - Happy in castlemilk? Deprivation and depression in an urban community. AB - This paper reports findings from a survey conducted in Autumn 1993 in Castlemilk, Glasgow. The purpose of the survey, commissioned by local service providers, was to estimate the prevalence of sub-clinical depression, identify at-risk groups and to specify how the interplay of different factors impacts on people's mental health. Just under a quarter of residents experienced some emotional distress. The factors associated with the observed distress were located both within the biography of those surveyed and the structural features of the locality in which they lived. It is concluded that service providers can devise strategies to identify individuals at risk of emotional distress to help them confront their private troubles. These should however be supplemented at regional and national levels by a range of policies designed to address structural inequalities. PMID- 10670968 TI - Consumerist ideology and the symbolic landscapes of private medicine. AB - The consumption of health care has generally been examined by geographers in terms of patterns of service utilization by patients. The sites of service provision have been viewed as locations rather than as contributors to, and constituents of, urban landscapes. In this paper we argue that perspectives of the so-called "new" cultural geography can assist in interpreting the urban landscapes of health care. The case of accident and medical clinics in New Zealand is examined and used to argue that the underlying ideologies of private provision and consumerism is reflected in the symbols used in both the built environment and in advertising. PMID- 10670969 TI - Spatial distribution of watery diarrhoea in children: identification of "risk areas" in a rural community in Bangladesh. AB - To assess the geographic variation of acute watery diarrhoea in children 0 to 5 years old in rural Bangladesh, all cases of "cholera-like" diarrhoea were plotted on the map. A clustering pattern was noticed, and validated by a nonparametric clustering test for in-homogeneous population. Several risk areas for the disease were identified. In these areas, the point prevalence (8.7/1000) was notably higher than outside the areas (0.41/1000), (95% confidence interval, 15.55 29.30). Parents' education, population density and use of sanitary latrines were significantly related to the risk areas (p < or = 0.001). The results of this study indicate that computer-assisted mapping may be useful in defining and monitoring risk areas for watery diarrhoea in children. PMID- 10670970 TI - Stigmatization, HIV/AIDS, and communities of color: exploring response to human service facilities. AB - HIV and AIDS are rapidly becoming leading causes of death for men and women in large cities across the US. Epidemiological data indicate that persons of color in particular have been disproportionately affected by HIV/AIDS. The continuing growth in the incidence of HIV/AIDS among persons of color implies that human service facilities will be needed in close proximity. However, there has been little research exploring response to human service facilities associated with HIV/AIDS in communities of color. This paper explores community response to facilities associated with HIV/AIDS by analyzing in-depth interviews with fifteen Vietnamese and Latino/Latina informal opinion leaders in Orange County, California. These interviews indicate that the stigma surrounding HIV/AIDS emanates to a large degree from the social construction of "HIV/AIDS as homosexuality". Even with the deviance and marginalization associated with HIV/AIDS, however, creative coping strategies have been developed by families within the Latino and Vietnamese communities to enable the maintenance of family ties with persons living with HIV/AIDS. PMID- 10670971 TI - Uncertainty, reassurance and pollution: the politics of epidemiology in Teesside. AB - This article focuses upon longstanding controversy surrounding the health impact of air pollution from steel and chemical industries in the conurbation of Teesside, in Northeast England. Drawing on Ulrich Beck's analysis of risk, it presents a commentary on the ways in which epidemiological findings are incorporated or marginalised in conflicting public health narratives in Teesside, taking recent epidemiological research on mortality patterns for detailed scrutiny. I argue that where public health issues are contentious and politicised, inconclusiveness in such research plays a significant part in sustaining a "narrative of reassurance" about the possible health or environmental costs of living close to industrial operations. PMID- 10670972 TI - Quantitative health research in an emerging information economy. AB - This paper is concerned with the changing information environment in the U.K. National Health Service and its implications for the quantitative analysis of health and health care. The traditionally available data series are contrasted with those sources that are being created or enhanced as a result of the post 1991 market-orientation of the health care system. The likely research implications of the commodification of health data are assessed and illustrated with reference to the specific example of the geography of asthma. The paper warns against a future in which large-scale quantitative health research is only possible in relation to projects which may yield direct financial or market benefits to the data providers. PMID- 10670973 TI - Urbanization and mental health in Brazil: social and economic dimensions. AB - While there have been attempts to examine urbanization and the quality of urban life as special risk factors in the generation of mental illness, the issue is controversial and remains largely unresolved. In this article particular consideration is given to the process of contemporary structural transformation in the Brazilian society leading to urbanization. Selected Brazilian studies of mental illness in urban areas are described. Economic displacement, or unemployment, emerges as the most significant risk factor for mental ill-health and is more important than, for example, rural to urban migration. Current studies are then criticized for focusing too much on the search for single risk factors instead of taking the social, structural context into account. PMID- 10670974 TI - The contribution of coterminosity to joint purchasing in health and social care. AB - This article examines the context within which coterminosity (the coincidence of geographical boundaries between two or more organisations) is currently being re defined, given the development of purchasing in health and social care agencies in the British welfare system. It explores the current trends within purchasing and especially the notion of "locality" as a means of promoting "joint working" between purchasing agencies. In particular, the emergence of general practice as a focus of purchasing is explored with reference to its involvement in joint purchasing (or joint commissioning) and its interaction with the "locality". The article concludes that coterminosity has a contribution to purchasing organisations but increasingly at a local level such as the general practice or locality. This local manifestation of coterminosity may minimise the effects of fragmentation and encourage inter-agency collaboration. PMID- 10670975 TI - Telecommunications and disabled people: a rural perspective. AB - In the context of the National Health Service and Community Care Act 1990 this paper focuses on the role of home- and community-based telecommunication services in sustaining the life-style and independence of elderly and disabled people in rural Britain. It draws on findings from a recent survey in the North Cotswolds to evaluate four models of telecommunication support: telephone trees; community alarm systems; community teleservice centres (CTCs); and on-line computer systems. The arguments in favour of a "low-cost-voluntary-care-support-model", a variant of the CTC, are discussed with respect to client need and community setting. PMID- 10670976 TI - Cattle markets and local festivals: development of HIV/AIDS prevention interventions for specific risk situations in rural northeast Thailand. AB - In rural Northeast Thailand, risk of sexual transmission of HIV is popularly perceived to be site-specific. Risk of HIV transmission in local scenarios like cattle markets and village festivals has not been adequately addressed. This paper assesses the use of community consultation and formative research to overcome prevailing assumptions about HIV risk by involving community members in the process of identifying risks and developing HIV prevention strategies. This participatory approach can be used to develop prevention programs that are responsive to the specific context of risk behavior in rural environments. PMID- 10670977 TI - Community pharmacy in South Africa: a changing profession in a society in transition. AB - The analysis of community pharmacy as a profession in transition acquires an additional dimension in South Africa, since it is inextricably linked to its social characteristics as well as to the political transformation taking place. Using data collected by means of a documentary search, interviews with key informants and a survey of community pharmacists, the paper presents the relevant societal features and explores some of the complexities associated with the existing as well as the potential future role of community pharmacy in the context of changing health services in a society in transition. It concludes that the changes in community pharmacy and the role it can play in the provision of Primary Health Care to all the people of South Africa are linked to the greater transition in society and its future health care services. PMID- 10670978 TI - Therapeutic landscapes of the Jola, The Gambia, West Africa. AB - This paper contributes to the 'new' medical geography through its analysis of the therapeutic landscapes of the Jola of The Gambia. The paper advances the debate surrounding the conceptualization of medicine and health through a review of literature on African medicinal systems; it examines in detail the health care system of the Jola of The Gambia, documenting indigenous human and ethnoveterinary medical beliefs and practices and focusing in particular on the role of herbal medicine; and it discusses the interactions and links between indigenous medicine and biomedicine, thus demonstrating the importance of placing an understanding of health care systems in different places within an awareness of global power relations. The paper therefore links cultural perspectives with a political economy analysis, to highlight the importance of place and specificity of cultural context when investigating health care beliefs and practices. The intention of the paper is to present a theoretically informed empirical case study which reinforces the practical value of a 'new' medical geography. PMID- 10670979 TI - Talk about cancer: environment and health in Oceanpoint. AB - Based on an ethnographic study, this paper explores one Australian community's 'popular epidemiology' of the role of the environment on health. Residents express concern about cancer risks due to contamination from a land-fill dump site and from radioactivity from previous mining activities. These concerns carry a range of meanings, not only regarding the threats to the physical place of Oceanpoint, but the social space the community occupies and the values they espouse. The perceptions of health risks in this location involve a contest over the cultural politics of place, the political economy of development and the sense of a former authentic 'community' that is being lost and disempowered from controlling its future. PMID- 10670980 TI - Moving experiences: a qualitative analysis of health and migration. AB - This paper describes a qualitative analysis of the health and health care experiences of South Asian Fijian women now living in the lower mainland area of British Columbia, Canada. A particular focus is put on the health impacts of the migration experience. A thematic analysis of in-depth interviews informs the discussion of individual women's, as well as service providers', views of health meanings, physical and emotional health concerns, experiences with the health care system, and women's roles as care-givers. The findings have implications for how health and illness are conceptualized, and how health services are provided to particular groups in particular places. PMID- 10670981 TI - The politics of methodology in 'post-medical geography': mental health research and the interview. AB - This paper argues that emerging 'post-medical geographies' require attention to the methodological in order to fully appreciate how different geographical knowledges are produced and contextualized within the politics of research relationships. 'Geographies of mental health and illness' are focused upon in order to argue that the 'peopling' of health research should also be accompanied by debate about what sorts of methodologies we employ in accessing these minds/bodies and voices. The research interview is a primary focus here. A critique of psychoanalytic approaches to geographical research argues that such 'models' of interpretation and management can mean that participants or research 'subjects' can be framed in almost diagnostic categories of behaviour. Empirical examples of mental health research in Nottingham are used to argue that more flexible approaches which pay attention to perceived dualisms (such as 'sanity' and 'insanity'), negotiation, embodiment, socio-spatial contexts and content within the interview situation may aid in understanding the politics which encompass geographical health research. PMID- 10670982 TI - Mortality from non-Hodgkin lymphoma and UV exposure in the European Community. AB - There has been a large, unexplained rise in the incidence of non-Hodgkin's lymphoma (NHL) in many countries. It has been hypothesised that increased exposure to solar ultraviolet (UV) radiation may have been a factor in this increase. The hypothesis that exposure to ultraviolet radiation is a factor in NHL can be tested by examining whether geographical variations in UV and in the incidence of the disease are positively correlated. Previous studies have given mixed results but some of these have failed to take into account confounding by socio-economic factors and the multilevel structure of data derived from several different countries. It was therefore decided to carry out a study using data on NHL mortality for the period 1971-80 for level II administrative units in 9 countries in the European Community. Estimated levels of solar UV and per capita GDP were also derived. Poisson regression models of the relationship between NHL mortality, UV and per capita GDP, taking into account the multilevel structure of the data, were fitted using the MLn package. Simple models that did not adjust for the effects of variations in per capita GDP or account adequately for the structure of the data produced apparent negative or quadratic associations between NHL and UV. However, further models show that there is a highly significant positive association between NHL and per capita GDP. Once this is included in the fixed and random parts of the multilevel model the association between NHL and UV becomes positive although non-significant (p = 0.081) at the conventional 0.05 level. These results underline the need to control for socio economic factors and to take into account the multilevel structure of the data. Studies using international data that do not do this run the risk of producing misleading results. PMID- 10670983 TI - Advice-giving in community pharmacy: variations between pharmacies in different locations. AB - The advice and services provided by community pharmacies are viewed by policy makers as having an increasingly important contribution to make as a primary health care resource to local populations. However, little attention has been given to the variations which may exist between pharmacies operating in different localities. Findings from an ethnographic study of pharmacies illuminate differences in the nature and quality of advice and services provided by pharmacies operating in disparate localities. Analysis of qualitative data suggests that differences in the environment within which pharmacies are located and organised influence the type of service provided to local populations. The possibility of an inverse care law operating in relation to the nature of services in poor urban localities compared to those in rural areas is also discussed. PMID- 10670984 TI - Credentialling immigrant physicians in Israel. AB - Credentialling of immigrant physicians is discussed in the context of two sets of high-priority values which have remained constant in Israel since its founding: (a) an open, non-selective migration policy which has resulted in the entry of thousands of immigrant physicians, (b) the high priority accorded to quality health care. These values and their social implications are discussed in terms of the licensing procedures before 1988 and after that date when more stringent procedures were initiated. These processes are discussed with special reference to the large number of immigrant physicians that have come to Israel from the former Soviet Union since 1989. PMID- 10670985 TI - The geography of dementia: an approach through epidemiology. AB - Dementia is an extremely common condition among the elderly. Over 100 epidemiological surveys have now been published from many countries. Methodological differences make comparisons difficult, although meta-studies carried out in Europe provide baseline material for developed countries. Surveys within each geographical area which has been studied are discussed. Differences in the distribution of sub-types of dementia and the resultant challenges to governments and researchers are set out. PMID- 10670987 TI - UK and western European late-age mortality: trends in cause-specific death rates, 1960-1990. AB - Age, sex and cause-specific death rates for the elderly population of 16 western European countries are examined for 1960, 1970, 1980 and 1990. Over the 30 years, the all-cause rates have fallen by around 23-41% depending on age and sex. Mortality from stroke has declined substantially and from cardiovascular disorders has recently fallen, but cancer health rates have increased among men. A comparison of the UK death rates with the west European and Swiss rates finds relative improvement in the UK for male mortality, but that female mortality at the younger ages has worsened sharply. Cardiovascular and stroke mortality is now exceptionally high in the UK among females aged 60-64 years and the 1980s trends for the 60-64 and 70-74 years age groups were unfavourable for several other causes of death. PMID- 10670986 TI - Home is where the governing is: social capital and regional health governance. AB - The relationship between the civic nature of a community and effective political governance by regional health boards in Canada is explored. A model is proposed that identifies components of social capital such as trust, commitment and identity, associationalism, civic participation and collaborative problem solving. These concepts are then theoretically linked to effective governance, in particular to reflection of health needs, policy implementation, population health, fiscal responsibility and administrative efficiency. The generalizability of this model is discussed, as are current research directions and policy implications for governments. The conclusion is that governments might want to incorporate a dual perspective encompassing both the political institutions and the community structure. PMID- 10670988 TI - A death in our street. AB - How do neighbours relate to the dying, and what effect does a death have on a neighbourhood? After briefly reviewing related literature, the article explores the concepts of reciprocity and solidarity, using the recent work of Michael Young to highlight limitations in the work of Philip Abrams. The potential of suffering to generate neighbourhood solidarity in an increasingly affluent and private society is considered. PMID- 10670989 TI - Words in wards: language, health and place. AB - The role of place in medical encounters which involve language is examined using theoretical arguments backed by empirical studies. Links between language and place, health and place and especially language and health are discussed. The language-health link is developed in terms of explanatory models; how language is used in medical encounters; and power, dominance and resistance relationships. Then it is shown how considerations of place enhance knowledge about this link. The paper closes with a set of research questions which focus on the role of place. PMID- 10670990 TI - Guthrie house: a rural community organizing for wellness. AB - This paper is interested in the issue of community participation and empowerment in health care provision and decision-making. In Canada, the present scope for public involvement in planning or managing the state's health and social services system is limited. This poses a particular problem for rural communities--places where the provision of health care services has historically been limited when compared to urban locations. These rural communities are now facing a double burden as public policy moves increasingly towards a retrenchment of the welfare state. This paper examines one rural community's response to this double burden. The village of Elgin in rural Ontario recently established Guthrie House, a community-based resource center for health and wellness services. Community participation in this case involved a level of control whereby local citizens together defined the health and social care services that they saw as best meeting the needs of their community. This form of community participation is considerably different from the forms of public involvement in the established medical system and represents a critical link to 'empowering' the local community as partners in health care. Through an examination of Guthrie House, the paper presents a review of some critical 'characteristics' which mark successful community self-help organizations and concludes with a discussion of the policy implications for greater community participation. It is argued that such community participation in health care is a policy option which government should be paying particular attention to in these times of fiscal constraint, increasing health care needs and increasing consumer dissatisfaction with government service provision mechanisms. PMID- 10670991 TI - National identity and controversy: New Zealand's clean green image and pentachlorophenol. AB - In the 1990s regulatory bodies in New Zealand worked to develop guide-lines to clean up the contamination of land caused by the use of pentachlorophenol in the treatment of timber. In contrast, there has been little effort to identify and compensate workers contaminated at these sites. This paper explores some of the reasons why action over the contaminated land was relatively quickly taken, whilst there was a lingering controversy over the health of the workers who used pentachlorophenol. The case study suggests that symbols of national identity can play an important role in the resolution of controversy. PMID- 10670992 TI - Primary care needs assessment and resourcing: complementary practice and geographic perspectives. AB - This paper considers the use of small area census data in deriving socio-economic health need indices for primary care practices and assesses how far such indices are related to practice activity. A particular goal is the derivation of weightings for resourcing formulae which incorporate the effect of socio-economic deprivation on activity as well as the more usual allowance for patient age alone, so ensuring a closer match between needs and resources across both practices and localities. A case study is undertaken of a health authority in East London with a population of 380 thousand and 97 primary practices. Total rates of inpatient and outpatient referral and of prescribing by practices all show an influence of practice deprivation after allowing for age, and this effect is enhanced for subcategories such as emergency referrals to A&E departments. After taking account of practice age structure and deprivation, practice characteristics themselves (e.g. number of GPs in relation to practice population) are relatively unimportant influences on activity. The paper discusses how variation in need may be incorporated into resourcing via differential weighting. PMID- 10670993 TI - Is there a geography of alternative medical treatment in The Netherlands? AB - An increasing number of people are using alternative medical care. The literature suggests that there are important between place variations, however. This paper tries to assess the extent of these variations and mechanisms behind them for the utilization of homeopathy, paranormal healing and manual therapy. Are these variations a matter of level of supply, degree of urbanization, GP characteristics or simply a matter of composition of populations? Data are derived from the Dutch National Surgery of General Practice and analyzed using multilevel logistic regression models. Between place variation in utilization of homeopathy is mainly a matter of composition of populations with respect to health locus of control and religion. With respect to paranormal healing, it is exclusively a matter of religion. With respect to manual therapy, place variations are a matter of individual, GP, as well as area characteristics, but a relatively large amount remains unexplained. PMID- 10670994 TI - Trauma systems and major injury centres for the 21st century: an option. AB - Trauma, especially involving accidents and falls is now the leading cause of death in the UK in the first four decades of life. This paper looks at the existing pattern of Accident and Emergency Units and suggests that because it has grown up in a fragmented and poorly co-ordinated fashion and because it is subjected to demands that it was not designed for, it is inadequate in dealing with the most severe injury cases. Following collaborative work with researchers at the North Staffordshire Hospital, the authors propose a system of thirty major trauma centres to cover the whole of the UK. Each would deal with the most severe accident cases from surrounding hospitals. Using data from the census and from health authorities, analysed within a GIS environment, a locational pattern for these centres is suggested and their catchment areas are outlined. PMID- 10670995 TI - Melanoma incidence in Connecticut by town of residence. AB - In an analysis of 2,935 cases of invasive melanoma of the skin diagnosed among residents of Connecticut's 169 towns in 1990-1994, 12 of the 17 towns with statistically significantly elevated standardized incidence ratios (SIRs) were on or near the ocean shoreline (in the extreme southern part of the State). The distribution of melanoma thickness did not support the hypothesis of greater detection of clinically less significant cancers in the ocean shoreline area with high SIRs. In multiple regression analyses, town location (on the ocean shoreline) was a statistically significant predictor of SIR by town, independent of socioeconomic indicators and racial composition. Although the effects of shoreline residence and latitude were difficult to separate, the results should be useful in planning analytic epidemiologic (i.e. case-control) studies in specific geographic areas for testing etiologic hypothesis as a basis for planning public health interventions. PMID- 10670996 TI - Environmental health metanarratives: an analysis of policy making in Ontario, Canada. AB - This paper provides a narrative analysis of a policy document, issued by the Ontario Premier's Council on Health, Well-Being and Social Justice, entitled "Our Environment, Our Health". It begins by outlining the background for the production of the document and establishing the nature of narrative analysis. The intention of this method is to reveal dominant and suppressed ideas through identifying narrative, non-stories and counter-stories. It then goes on to provide an analysis of the overall policy piece and its constituent parts, healthy ecosystems, healthy communities and healthy workplaces. The document's power is seen not only in its authorship but also in demonstrating the moral basis of individual responsibility to achieve collective targets and to undertake collection action. PMID- 10670997 TI - Assessing ecologic proxies for household income: a comparison of household and neighbourhood level income measures in the study of population health status. AB - This paper examines the validity of using ecologic measures of socioeconomic status as proxies for individual-level measures in the study of population health. Based on a representative 5% sample of households in a Canadian province, the study integrated three sources of information: administrative records of individual health care utilization, records of deaths and 1986 census records which contained information on household income and average neighbourhood income. Thirteen measures of health status were developed from these sources of information. The hypothesis that risk estimates derived from ecologic income measures will be attenuated relative to estimates obtained from household income was not supported. These results provide evidence for the use of ecologic-level measures of income in studies which do not have access to individual-level income measures. PMID- 10670998 TI - Subject loss in spatial analysis of breast cancer. AB - Possible selection bias from assignment of latitude-longitude coordinates to the place of residence of all Connecticut women diagnosed with breast cancer from 1992 to 1995 (N = 11,470) was evaluated. Exact address-matching was accomplished for 8,121 records (70.8%) and an additional 1,722 records (15.0%) were matched using relaxed criteria. We did not address-match 1,627 records (14.1%) due to missing address information or limitations of the Geographic Information Systems street file. The age-adjusted likelihood of address-matching records was significantly greater for women of color, those born within Connecticut, residents of urban locales or census tracts with low median family incomes and those cases diagnosed nearer to 1992. Few differences in address-matching were attributable to tumor characteristics or therapeutic modality. PMID- 10670999 TI - Sociodemographic and geographic patterns of firearm suicide in the United States, 1989-1993. AB - This study identified the sociodemographic and geographic patterns of using firearms to commit suicide in the United States. Data from the Mortality Detail Files (1989-1993) were analyzed using logistic regression. The adjusted odds of using firearms increased with age among men and decreased among women. Widowed men and married women had the highest odds of using firearms. The odds were highest among those without college education, in nonmetropolitan areas and in the East South Central and West South Central geographic divisions. The likelihood of using firearms to commit suicide varies significantly across sociodemographic and geographic subgroups of the US population and parallels patterns of gun ownership. The results of this study suggest that regional cultural factors play an important role in accounting for the differential rates in suicidal behavior involving firearms. PMID- 10671000 TI - The decrease of life-expectancy at birth in Romania: some crisis contributing factors. PMID- 10671001 TI - Behind the crumbling walls; the re-working of a former asylum's geography. AB - This paper attempts to examine the effects of deinstitutionalization on a former asylum's human and physical geography, through a case study of St Lawrence's Hospital, Bodmin, UK. It researches the inevitably impermanent geography of an asylum in the process of being 'run down'. The paper includes an empirical account of the rationalization of a former asylum, reconstructing an east to west migration of activities across the site. It deals with St Lawrence's as a 'stigmatized' place unable to escape the mark of its asylum past, and as a place that staff and local people cannot imagine being used for anything other than land uses associated with 'mad people' or other marginal groups in society. The paper deals with how St Lawrence's is regarded by some as a 'community', people who recall with a measure of affection how the asylum used to be a 'home' as well as a 'workplace', a nostalgia which rebounds into suspicion about the dismantling process. Fundamentally, the paper aims to tease out and work through the hopes and fears that now surround the former asylum's environment. PMID- 10671002 TI - Integrating space into a reactive theory of the asylum: evidence from post-Civil War Georgia. AB - Absent from the geographical literature on the 19th century asylum is a reactive theory of social control. Such an approach focuses on the importance that families and communities of the insane played in guiding institutionalization. Thus far, reactive theory has emphasized how social distance within communities shaped public reaction to mental illness and hence the admission decision. Unfortunately, it ignores the importance of spatial distance from the asylum in molding utilization patterns. Using evidence from asylum admissions in post-Civil War Georgia, this paper explores the necessity of adopting a reactive theory and the even greater necessity of integrating space into reactive theory. Such a project requires going beyond traditional treatments of spatial distance as a geometric barrier and examining the politics of distance, that is, how the meaning of journey to the asylum was open to competing and conflicting constructions. PMID- 10671003 TI - Sentenced in sorrow: the role of asylum in the Jean Gianini murder defence. AB - This paper describes the role played by the notion of asylum in the legal battle over confessed murderer Jean Gianini's mental competence and commitment. Gianini was a 16 year old who murdered his former teacher in a small upstate New York town in 1914. His trial was the first in the US to employ the Binet-Simon intelligence test as a defence and featured a clash of expert witnesses whose credibility was based upon their residence and work in asylums. The verdict of 'not guilty due to criminal imbecility' was due to the defence team's successful portrayal of the asylum as a punishing prison from which the defendant would never be released. PMID- 10671004 TI - 'She thinks this is the Queen's Castle': women patients' perceptions of an Ontario psychiatric hospital. AB - Drawing on the detailed information contained in patient case files, this study explores women patients' perceptions of the Ontario Hospital, Cobourg, a psychiatric facility for women which operated from 1920 to 1974. The paper concludes that, while twentieth-century psychiatry promoted an image of asylums as modern medical facilities, women patients continued to view these institutions as 'crazy houses', 'havens' and 'prisons'. It was not until the 1950s and 1960s, the eve of the decline of the authority of scientific psychiatry, that some women patients began to conceive of psychiatric hospitals as 'medical' facilities where therapeutic care could be sought. PMID- 10671005 TI - Across the water: reviewing geographical studies of asylums and other mental health facilities. AB - It is possible now to identify a small field of geographical studies exploring how space, place, environment and landscape are bound up in the worlds of people experiencing mental health problems. Some of these studies take seriously the institutions which have been provided to shelter, control, care for and even cure such people, and this interest has often touched upon the rise of the 'asylums' in Europe and North America (chiefly as an eighteenth- and nineteenth-century phenomenon). This paper surveys the geographical literature tackling asylums and other mental health facilities, and then offers an interpretation of the theoretical claims and substantive research undertaken in this respect by Michael Dear and various co-workers. Running through the paper is an argument about the need for studies of 'asylum geographies' to be attentive to what Gunnar Olsson terms 'ontological transformations' between thoughts and things. PMID- 10671006 TI - Moral architecture: the influence of the York Retreat on asylum design. AB - Institutional and architectural history places the asylum alongside the prison and other institutional types whose architectural characteristics emphasized confinement and control. This history obfuscates important differences in how ideas about treatment were represented in the particular design of these institutions; in other words, how the structure of a place became part of its discourse. What becomes obvious in nineteenth-century, asylum architecture is the influence of a small Yorkshire private asylum built by a Quaker, William Tuke, in 1796. The York Retreat, in form, solidified the ideas of 'moral treatment' in design and in turn assumed an exalted character in the design of late nineteenth century asylums. Every researcher working in the field of the history of insanity acknowledges the importance of this event and its impact on the discourse of insanity for the century to follow. Few however talk about how its unique design was incorporated as part of this discourse. PMID- 10671007 TI - The geography of mental health in Dunedin, New Zealand. AB - The process of deinstitutionalization of people with chronic mental disabilities in Western countries has often produced a spatial concentration of ex-psychiatric patients, and of mental health services, in inner city urban neighbourhoods. In this paper, the geography of mental health services and patients in Dunedin is examined, and it is shown that a concentration does exist in one neighbourhood. The history and characteristics of this neighbourhood are described. The key factors in contemporary New Zealand that have generated this spatial pattern are then considered, and Dunedin's centralized mental health geography is contrasted with the North American "zone of dependence" phenomenon. The paper concludes by considering to what extent Dunedin's emergent geography of mental health provides a supportive environment for people with mental illnesses, and exploring the policy implications for health care planners and service providers. PMID- 10671008 TI - Geographical change in residential care provision for the elderly in England, 1988-93. AB - This paper offers an investigation of the spatial consequences of changes in the structural organization of residential care in England between 1988 and 1993. Data from various government publications were analysed using descriptive and spatial statistical methods. While the study period witnessed an overall levelling of residential care growth, the independent (i.e. private and voluntary) sector's share of all elderly residents in England increased from 56% to 73%. At both national and intra-regional scales, the structural changes resulted in an increasing geographical concentration of public sector residents and a moderate trend towards a more uniform spatial distribution of private residents. PMID- 10671009 TI - Dementia and help with household tasks: a comparison of cases and non-cases. AB - This paper is based on data, relating to people aged 75+ living in the community, from the city of Liverpool and from a rural area of North Wales. It compares those suffering from cognitive brain disorders with those identified as being cognitively unimpaired. Levels of need, and sources of help with a range of domestic and home maintenance tasks are identified. Results show that most help for those who are cognitively impaired comes from relatives living in the same household, while help for those who are physically impaired comes primarily from spouses or relatives living in different households. Cases do not demonstrate a higher level of use of formal services than non-cases. Implications for policy and practice are discussed. PMID- 10671010 TI - Gendered health policies and a women's movement: the Gypsy case. AB - Health concepts depend on socially constructed hygiene practices. Many accounts of "traditional" Gypsy hygiene make gender differentiation primary, implying that traditional health practices reinforce patriarchy and emphasizing the special needs of Gypsy women. This paper argues, however, that the position of women is contested in Romani society. Although prioritizing the more apparent health needs of women and small children appeals to health professionals, this cannot by itself create an effective critique of the health system's response to Gypsy needs. The emergence of a Gypsy women's movement has led to a new critique, and challenge to the victim image of women. PMID- 10671011 TI - Nineteenth century Canada: indigenous place of dis-ease. AB - During the nineteenth century period of intensive European Expansion into Canada, place was experienced with dis-ease by indigenous people. Not only was there less land available for people of the First Nations to live on as in the past centuries, but their intimate relationship with the land was disturbed causing a dis-ease, as their ability to experience place through ceremony was denied. The effects of this process of Euro-Canadian invasion within Canada created a sense of dis-ease, a sense of being out of place for indigenous people. It is my contention that in order to understand health, an awareness of the dynamic connection between a people and their land is needed, recognizing that a quintessential human quality is to imbue the world with meaning in the creation of sense of place. Through examination of a variety of published and unpublished material, archival sources, memos, letters, official documentation, I explore the effects of control of the place of First Nations, on the health and healing practices of the indigenous people. PMID- 10671012 TI - An entropy-based algorithm for detecting clusters of cases and controls and its comparison with a method using nearest neighbours. AB - A new method for detecting disease clustering based on entropy is presented. For this method cases and controls are plotted on a map. The map is divided into regions. The entropy of the space is calculated as the log of the number of possible ways of placing the cases and controls in the various regions given the total number of cases and controls and the number of cases and controls in each region. The power of the entropy technique is tested against the power of the nearest neighbour technique (NNT). The entropy method is shown to be substantially more powerful than the NNT when there is more than one cluster in the space or when the clusters are near the boundary of the space. PMID- 10671013 TI - Allocating resources in health care: alternative approaches to measuring needs in resource allocation formula in Ontario. AB - Maintaining or improving the welfare of the population is a complex issue involving individual and collective actions and institutions. Despite questions regarding the relevance of health care systems to these aims, they remain vital policy and treatment arenas with respect to curative and preventative regimes. As a component of social welfare, health care resources should be distributed equitably, according to need for health care. This paper evaluates alternative indicators of health status within Ontario against self-reported health as a means of allocating health care resources. Proxies of need for health care include standardized mortality ratios (based on the population aged 0-64) and a socioeconomic based indicator. Mortality indicators are found to be more closely correlated with self-reported health status than the socioeconomic indicator, suggesting that mortality is better able to reflect variations in health status and health care needs. PMID- 10671014 TI - Social and local variations in the use of urban neighbourhoods: a case study in Glasgow. PMID- 10671015 TI - Health and nature in the 19th century Australian women's popular press. AB - This paper asks how health and nature are represented in the Australian women's press during the late nineteenth century. A time of significant social change during which women, and sympathetic male colleagues, challenged traditional roles as pathological creatures of the domestic sphere, this period is explored through the writing of women working for popular magazines. As women captured, transformed and redeployed stereotypical views of them as essentially and naturally ill, they consolidated their push into the public realm, while also convincing themselves and others of their vital place in the private sphere, but as capable, well and fit creators of people and of a nation. PMID- 10671016 TI - Explaining geographies of health care: a critique. AB - This paper considers the ways in which geographers have sought to explain the spatial organisation of health care services. It does so at three interlocking scales: the global/international, the national, and the local. It considers the substantive adequacy and explanatory problems associated with different perspectives and also discusses the normative implications of alternative interpretations of patterns of health care services. The paper notes the ways in which some conventional geographies of health care, which seemed to postulate convergence towards greater egalitarianism in service provision between and within states, have been challenged by changing economic circumstances, and by a changing political and intellectual agenda. The paper also considers some emerging geographies of community-based struggles around health services and discusses their potential and limitations. Finally there is a discussion of the potential contribution, if any, of a distinctively geographical perspective on health care. PMID- 10671017 TI - Modernization and health care in contemporary China. AB - This paper provides an analysis of the changes that have occurred in the Chinese health care system in the reform era, post-1978. The economic reforms have brought significant benefits to many Chinese people, but the benefits have not been distributed evenly throughout Chinese society. The paper evaluates some of the evidence indicating deterioration in both the quality and quantity of health care services in the reform era. There is some alarming evidence that rural women, especially those living in the country's poorest regions, have been experiencing lower levels of accessibility to health care, and increasing levels of both morbidity and mortality. It is also evident that during the reform era the gap between service levels in rural areas and the cities has widened, and that chronic poverty still exists in large parts of the countryside. It is important to stress that these inequalities were not created by the economic reforms--in fact they were first observed during the supposedly egalitarian era associated with Mao Zedong. The economic reforms and the widespread implementation of market socialism in Chinese society, have, in general, been associated with improved health status and greater access to health care. It is also apparent, however, that pre-existing inequalities have not been eradicated by the reforms, and, particularly along the lines of gender and geography, the evidence points to a widening of inequality. PMID- 10671018 TI - Does housing tenure predict health in the UK because it exposes people to different levels of housing related hazards in the home or its surroundings? AB - In the UK housing tenure (whether the dwelling is owner occupied or rented) has consistently been found to be associated with longevity and with a number of measures of health. It has been argued that it is a good measure of material circumstances, and it is often incorporated into area based measures of social or material deprivation. However there is little published research on whether housing tenure predicts mortality and morbidity simply because it is an indicator of material well being, or whether, in addition, different categories of housing tenure expose people to different levels of health hazards in the dwelling itself or in the immediate environment. In this paper we examine, using data on adults aged 40 and 60 from socially contrasting neighbourhoods in Glasgow, Scotland, whether housing tenure is associated with housing stressors (e.g. overcrowding, dampness, hazards, difficulty with heating the home) and with assessment of the local environment (e.g. amenities, problems, crime, neighbourliness, area reputation and satisfaction), and whether this might help to explain tenure differences in long-standing illness, limiting long-standing illness, anxiety and depression. Controlling for income, age and sex, housing stressors independently predicted limiting long-standing illness; assessment of the area and housing type independently contributed to anxiety; and housing stressors, housing type and assessment of the area independently contributed to depression. This suggests that housing tenure may expose people to different levels of health hazards, and has implications for urban housing policies. PMID- 10671019 TI - Community characteristics as predictors of perceived HMO quality. AB - We model the impact of community characteristics on people's perceptions of the quality of their health care experiences in HMOs. We focus on three community characteristics: sense of community, population density, and population diversity. Sense of community refers to people's perception of interconnection, shared responsibility, and common goals. Population density and population diversity are community characteristics that affect transactions costs in terms of time and energy, and affect people's perceptions of their community. We use data from a 1993 Florida poll to estimate the relationship between HMO members' perceptions of problems with health care experiences (cost, choice, access, satisfaction) and community characteristics. We find that all three community variables are significantly associated with perceptions of health care problems. We also find that effects of community variables operate differently for those in HMOs vs. those under traditional insurance. This study is consistent with research showing that community characteristics impact the health status of community institutions. Results suggest that providers may be able to improve care by being more responsive to individuals' need for community, that providers and communities can mutually gain by collaborating to improve community health, and that it may be cost-beneficial to factor community issues more strongly into health care policy. PMID- 10671020 TI - Diminishing regional contrasts? The east-west-divide in health behaviour among Finnish adolescents. AB - We studied whether differences in health behaviour between Eastern and Western Finland have changed between 1977 and 1993 and whether adjusting for socio economic characteristics changes the relationship between region and health behaviour. The Adolescent Health and Life-style Survey data have been collected biannually by mailed questionnaires from nationally representative samples of 16- and 18-year-olds (n = 27,785). The response rate varied from 79 to 83% by year. According to logistic regression analyses, smoking and physical activity were as prevalent and changes over time similar in both regions. In alcohol, high fat milk and female butter use the East-West-difference persisted over time. Regional differences were mainly explained by socio-economic variation between the two regions. PMID- 10671021 TI - African mortality and the new 'urban penalty'. AB - This paper reviews trends in rural/urban under-5 mortality differentials in Sub Saharan Africa in historical perspective, with particular attention to the case of Kenya. The rural/urban mortality gap has narrowed within the last half century, but while this was largely due to rapidly falling rural infant and childhood mortality over most of the period, in recent years it has been due primarily to a stalling and even upturn in urban under-5 mortality as urban economic and environmental conditions have sharply deteriorated in rapidly growing cities. Policy attention and resources need to be directed to large urban areas to prevent further deterioration of urban mortality and associated health conditions. PMID- 10671022 TI - Measuring access to primary medical care: some examples of the use of geographical information systems. AB - This paper explores the potential for geographical information system technology in defining some variables influencing the use of primary care medical services. Eighteen general practices in Scotland contributed to a study examining the accessibility of their services and their patients' use of the local Accident and Emergency Department. Geo-referencing of information was carried out through analysis of postcode data relating to practices and patients. This information was analyzed using ARC/INFO GIS software in conjunction with the ORACLE relational database and 1991 census information. The results demonstrate that GIS technology has an important role in defining and analyzing the use of health services by the population. PMID- 10671023 TI - Violent death in young people in the city of Sao Paulo, 1991-1993. AB - This paper aims to describe the principal causes of violent deaths among young people in the city of Sao Paulo, Brazil. Data from routine mortality statistics were used in the analysis. Young males were found to have a dramatically increased risk of death from violent causes especially those resident in lower income areas of the city. Possible explanations for these findings include economic instability generating social and cultural inequalities. PMID- 10671024 TI - Reply to Davis and colleagues: Twice weekly, directly observed treatment for HIV infected and uninfected tuberculosis patients: cohort study in rural South Africa. PMID- 10671025 TI - [Neuroprotection and cerebral ischemia. Symposium proceedings]. PMID- 10671026 TI - [Self-expanding prosthesis and esophageal cancer]. PMID- 10671027 TI - Professor Andre Collet 1945-1999. PMID- 10671028 TI - Current awareness in NMR in biomedicine. PMID- 10671029 TI - Mycoplasma pneumoniae-associated Henoch-Schonlein purpura nephritis. PMID- 10671030 TI - Ambulatory blood pressure monitoring for evaluation of hypertension in children. PMID- 10671031 TI - Oxalate nephrocalcinosis in renal tubular dysgenesis. PMID- 10671032 TI - Implementing clinical governance in the United Kingdom: the role of a regional haematology group. Northern Regional Haematologists Group. PMID- 10671034 TI - Selected papers from the 18th Annual Conference on Peritoneal Dialysis. Nashville, Tennessee, USA. February 1998. PMID- 10671033 TI - The geographic pattern of beta-thalassaemia mutations in the Portuguese population. PMID- 10671035 TI - British Society for Haemostasis and Thrombosis autumn meeting. Cambridge, United Kingdom, 22-24 September 1999. Abstracts. PMID- 10671036 TI - Inhibition of xanthine oxidase by flavonoids. AB - Various dietary flavonoids were evaluated in vitro for their inhibitory effect on xanthine oxidase, which has been implicated in oxidative injury to tissue by ischemia-reperfusion. Xanthine oxidase activity was determined by directly measuring uric acid formation by HPLC. The structure-activity relationship revealed that the planar flavones and flavonols with a 7-hydroxyl group such as chrysin, luteolin, kaempferol, quercetin, myricetin, and isorhamnetin inhibited xanthine oxidase activity at low concentrations (IC50 values from 0.40 to 5.02 microM) in a mixed-type mode, while the nonplanar flavonoids, isoflavones and anthocyanidins were less inhibitory. These results suggest that certain flavonoids might suppress in vivo the formation of active oxygen species and urate by xanthine oxidase. PMID- 10671037 TI - [Health promotion and health care in public health service of Bavaria. Proceedings of a meeting. Amberg, 19-21 October 1998]. PMID- 10671038 TI - Commentary: three decades of the inverse care law. PMID- 10671039 TI - Commentary: impact on health needs assessing from different angles. PMID- 10671040 TI - Decisions to withdraw treatment. Treatment can sometimes be withdrawn at home. PMID- 10671041 TI - Decisions to withdraw treatment. Courts can help resolve disagreement in difficult cases. PMID- 10671042 TI - Decisions to withdraw treatment. Most decisions are based on subjective appraisal. PMID- 10671043 TI - Incidence of venous thromboembolism in users of combined oral contraceptives. Risk is particularly high with first use of oral contraceptives. PMID- 10671044 TI - Partnership with patients. Local communities have role in influencing health policy. PMID- 10671045 TI - Partnership with patients. Modern antipaternalism needs to be invigorated. PMID- 10671046 TI - Partnership in patients. Perspectives must be reconciled. PMID- 10671047 TI - Partnership with patients. Doctors in Egypt deal with patients in their own way. PMID- 10671048 TI - Partnership with patients. Family doctors are part of team. PMID- 10671049 TI - Partnership with patients. Teamwork is necessary. PMID- 10671051 TI - Hyperbilirubinaemia in term infants. Hyperbilirubinaemia is a marker for inadequate breast feeding. PMID- 10671050 TI - Partnership with patients. A little knowledge can be a dangerous thing. PMID- 10671052 TI - Intervention for late life depression in residential care. How much trial and error should we tolerate in community trials? PMID- 10671053 TI - Intervention for late life depression in residential care. Important research seems to have been greeted with only two faint cheers. PMID- 10671055 TI - Diagnosing Lyme disease. Patients have to learn to help themselves. PMID- 10671054 TI - Intervention for late life depression in residential care. Cochrane preferred to use "effective" where other people used "efficacious". PMID- 10671056 TI - Intervention for late life depression in residential care. Cochrane may not have been first to define efficacy and effectiveness. PMID- 10671057 TI - Case report of a Thai male cystic fibrosis patinet with the 1898+ 1G-->T splicing mutation in the CFTR gene: a review of East Asian cases. Mutations in brief no. 196. Online. AB - Cystic fibrosis (CF) is the most common fatal autosomal recessive multisystem disorder, which occurs mainly in European-derived populations. The incidence of CF varies between 1 in 2000 to 3000 live-births in various ethnic groups. The disease is rare in East Asians. Here we report a 9 year old Thai male patient, who was diagnosed to have CF based on recurrent pneumonia, a slow weight gain, pancreatic insufficiency and repeatedly elevated sweat chloride levels by two different methods. A comprehensive genetic analysis showed the splicing mutation, 1898+ 1G-->T, which was apparently of maternal origin. Literature search found 39 documented cases of CF patients in East Asians. CFTR (MIM# 602421) genotyping was performed in 14 patients including our patient and in 9 of them a CF allele was identified. The findings seem to indicate that the splicing mutations, 1898+ 1G- >T and 1898+ 5G-->T are more common in East Asian CF patients. PMID- 10671058 TI - Mutation in the nerve-specific 5'non-coding region of Cx32 gene and absence of specific mRNA in a CMTX1 Italian family. Mutations in brief no. 195. Online. AB - Charcot-Marie-Tooth type I demyelinating neuropathies are genetically heterogeneous disorders (chrmosome 17,1,X). There are at least three genes on X chromosome, the more frequently involved being Cx32 in Xq13.1. Cx32 encodes for connexin-32, a gap junction protein of 283 aminoacids. We report the results of molecular studies in a CMTX1 Italian family, in which the mutation, found in the 5'-UTR, resulted in an abnormal mRNA connexin-32 expression. Mutations in PMP22 and P0 genes were also excluded in this family. Cx32 gene analysis carried out by PCR-SSCP on family members genomic DNAs, running a 321 bp fragment spanning the TATA box, the trasciptional start site, and the non coding exon 1b, revealed a shift correlated with a transition from C to T at position 40 of exon 1b of the 12 affected members, while was not found in the controls. Then the RT PCR-SSCP on cDNA from two peripheral nerve biopsies of two heterozygous females of the family were sequenced showing only the wild-type alleles and suggesting that mutated mRNAs were too unstable to be detected. The result also suggests a regulating role of the 5'-UTR of Cx32 mRNA. PMID- 10671059 TI - A novel mutation of the down-regulated in adenoma gene in a Japanese case with congential chloride diarrhea. Mutations in brief no. 198. Online. AB - Congenital chloride diarrhea (CLD) is an autosomal recessive disease characterized by excretion of watery stool with a high chloride content. Pathogenesis of CLD is a deficient absorption of chloride in exchange for bicarbonate in the ileum and the colon. In 1996, it was reported that 36 patients with CLD had mutations in the down-regulated in adenoma (DRA) gene; 32 Finnish patients had a three base deletion (951delGGT), 2 Polish patients had a one base mutation (371AtoT) and 2 Polish patients had a one base deletion (344delT). In this study we analyzed the DRA gene in a Japanese boy patient with CLD and in members of his family. The patient was found to have a two base deletion (TT) at nucleotide 1526-1527 within codon 509 which results in a frameshift leading to a permature stopping at codon 517. The patient was homozygous for the deletion, his parents and brother were heterozygous, and his sister was normal. This is the first case of CLD identified to carry a mutation of the DRA gene in Asia. PMID- 10671060 TI - A novel splice site mutation of the EXT2 gene in a Finnish hereditary multiple exostoses family. Mutations in brief no. 197. Online. AB - Hereditary multiple exostoses is a dominantly inherited disease characterized by multiple benign osteochondromas. The affected individuals have an increased risk of developing sarcoma. A large Finnish family with hereditary multiple exostosis was analyzed to find the disease-causing mutation. Blood samples were obtained from 35 family members, including 21 affected and 14 unaffected individuals. Using 2-point linkage analysis the EXT phenotype was shown to be linked to the recently cloned EXT2 gene on chromosome 11p11. The coding region of the gene was sequenced and a previously unreported splice site mutation found. This G to T transversion within a 5-prime splice donor site following exon 6 was shown to cause aberrant splicing of RNA. The described change is considered to be a novel disease-causing mutation in the EXT2 gene. PMID- 10671061 TI - Lys650Met substitution in the tyrosine kinase domain of the fibroblast growth factor receptor gene causes thanatophoric dysplasia Type I. Mutations in brief no. 199. Online. AB - Thanatophoric dysplasia (TD) is one of the most common neonatal lethal skeletal dysplasias with micromelic shortening of the limbs, relative macrocephaly, flat vertebral bodies and a narrow thorax. TD has been divided into two types, type I (TD1) and II (TD2), based on clinical, radiological, histological, and molecular criteria. We identified a A to T heterozygous base subsitution at position 1949 predicted to cause a Lys650Met substitution in a term infant with TD1. This mutation has been previously described in one case of TD1 and three cases of severe achondroplasia with acanthosis nigricans and mental retardation. Interestingly, all cases with the Lys650Met mutation have the same unusual curvature of the tibia and/or fibula. PMID- 10671062 TI - A genetic polymorphism in the human HOXB1 homeobox gene implying a 9bp tandem repeat in the amino-terminal coding region. Mutations in brief no. 200. Online. AB - In man there are 39 homeobox genes of the HOX family in four loci, HOXA, HOXB, HOXC and HOXD on chromosomes 7, 17, 12, and 2. We discovered the existence of two major alleles, termed a and b, of gene HOXB1. They differ at a specific position in the 5' portion of the coding region. Sequencing the two alleles revealed that the allele HOXB1A, contains two copies of the 9bp sequence 5'ACAGCGCCC3', starting at position 65 of the coding region, whereas the allele HOXB1b contains three copies of this sequence (Fig. 1). As a consequence, the allele HOXB1b encodes a homeoprotein containing two copies of the tripeptide HisSerAla, starting at amino acid residue 25, which is present in only one copy in allele HOXB1a. We analyzed 250 individuals and found that the allelic frequencies of HOXB1a and HOXB1b were 78.8% and 21.2%. The murine homologue contains only one copy of the 9bp repeat (Fig. 1). 7 mouse strains, namely 129, BALB/c, C57BL/6, C57BL/10, CAST/Ei, C3H and SPRET/Ei, are homozygous for this allele. The allele present in gibbon and rhesus monkey appears to be identical to the human HOXB1b (Fig. 1). PMID- 10671063 TI - A new nonamyloid transthyretin variant, G101S, detected by electrospray ionization/mass spectrometry. Mutations in brief no. 201. Online. AB - Familial amyloidotic polyneuropathy is caused by transthyretin (TTR) variants. The identification of new variants with and without amyloidosis may help to clarify the mechanism of amyloid fibril formation. We detected several variant TTRs from patients with and without symptoms of amyloidosis using mass spectrometry (MS). TTR was isolated by mixing test serum with anti-transthyretin antiserum, and the generated immunoprecipitate was analyzed by high performanced liquid chromatography/electrospray ionization (HPLC/ESI) MS. Variant TTRs showed extra peaks in addition to normal TTR peaks. A variant found in nonamyloid group was sequenced by HPLC/ESI tandem MS using peptides obtained by protelytic digestion of TTR and by DNA analysis. The structure was new, [G101S], and was found in a 74 years old Japanese male. This mutation results from substitution in a CpG hot spot. The substitution in the surface loop, 98-102, between F and G b strands may not cause amyloid formation. PMID- 10671064 TI - Hereditary nonpolyposis coloretal cancer: identification of novel germline mutations in two kindreds not fulfulling the Amsterdam criteria. Mutations in brief no. 203. Online. AB - Hereditary nonpolyposis colon cancer results from heritable defects in the MLH1, MSH2, PMS1 and PMS2 genes, which encode proteins involved in the mismatch repair process. In this work we report the identification of two novel germline mutations in the MLH1 gene from two unrelated HNPCC families. The two affected families do not fulfill the Amsterdam criteria. In family 1 we found a missense S93G mutation, which lies in a MLH1 domain critical for its MMR functions. In family 2 we found a two nucleotide insertion (AG) in position 523 from the AUG which determines an early stop codon at position 606 (codon 203). In both families the mutant alleles cosegregate with the cancer phenotype. PMID- 10671065 TI - Identification of 9 novel IDS gene mutations in 19 unrelated Hunter syndrome (mucopolysaccharidosis Type II) patients. Mutations in brief no. 202. Online. AB - Hunter syndrome is an X-linked lysosomal storage disorder caused by a deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). The IDS deficiency can be caused by several different types of mutations in the IDS gene. We have performed a molecular and mutation analysis of a total 19 unrelated MPS II patients of different ethnic origin and identified 19 different IDS mutations, 9 of which were novel and unique. SSCP analysis followed by DNA sequencing revealed four novel missense mutations: S143F, associated with the 562C-->T polymorphism, C184W, D269V and Y348H. Two novel nonsense mutations were found: Y103X (433C-->A) and Y234X (826C-->G). In two patients two novel minor insertions (42linsA and 499insA) were identified. In one patient a complete IDS deletion was found, extending from locus DXS1185 to locus DXS466). PMID- 10671066 TI - Mutations of the human tyrosinase gene associated with tyrosinase related oculocutaneous albinism (OCA1). Mutations in brief no. 204. Online. AB - Mutations in the human tyrosinase gene produce tyrosinase-related oculocutaneous albinism (OCA1, MIM #203100). Tyrosinase is a copper containing enzyme and is responsible for catalyzing the rate limiting step in melanin biosynthesis, the hydroxylation of tyrosine to dopaquinone. We report 13 new mutations in the tyrosinase gene associated with OCA1A (without pigment) and OCA1B (with pigment) including 9 missense mutations (H19Q, R521, R77C, G97R, C289R, L312V, P313R, F340L and H404P), two nonsense mutations (W80X and R116X) and two frameshift mutations (53delG and 223 delG). Our previous work has defined clusters of missense mutations that appear to represent functional domains of the enzyme, and three of the missense mutations fall into these clusters including two (F340L and H404P) that flank the copper B bindng site and the missense mutation R52I that is located in the amino terminal end cluster of the protein. The G97R missense mutation is the first identified within the epidermal growth factor (EGF)-like sequence and the H19Q missense mutation alters the cleavage site of the signal peptide sequence. Mutational analysis can provide a definitive diagnosis of the type of OCA as well as help structure/function analysis. PMID- 10671067 TI - Mutations of the human P gene associated with Type II oculocutaneous albinism (OCA2). Mutations in brief no. 205. Online. AB - Mutations in the human P gene lead to oculocutaneous albinism type 2 (OCA2, MIM #203200), the most common type of albinism in humans. The P gene encodes a 110 kDa protein that is associated with melonosomal membranes and contains 12 potential membrane spanning domains. The specific function of the P protein is currently unknown. We report 7 new mutations in the P gene associated with OCA2. This includes 6 missense mutations (S86R, C112F, A368V, T592I, A724P and A787V) and one frameshift mutation (1047del7). We also report 8 polymorphisms including one amino acid substitution, D/A257. We and others have found many polymorphisms of the P gene in the coding region, several of which result in amino acid substitutions, making molecular diagnosis problematic. In contrast to this is the tyrosinase gene associated with OCA1, with a limited number of polymorphic variations in the coding region. There is also no apparent clustering of P gene missense mutations in contrast to the clustering observed by the tyrosinase gene missense mutations that define functional domains of the protein. Further mutational analysis is needed to help define the critical functional domains of the P protein and to allow a definitive diagnosis of OCA2. PMID- 10671068 TI - Twelve novel RB1 gene mutations in patients with hereditary retinoblastoma. Mutations in brief no. 206. Online. AB - Hereditary predisposition to retinoblastoma is caused by germline mutations in the RB1 gene. Mutation analysis in this gene is important because knowledge of the causative mutation is often required for accurate risk prediction in relatives. We have performed RB1 gene mutation analysis in 45 patients with hereditary retinoblastoma. Screening by heteroduplex and SSCP analysis resulted in the identification of small mutations in 28 (62%) patients. Recurrent mutations, mostly CpG-transitions, were found in 16 patients. Two patients with isolated bilateral retinoblastoma showed missense mutations, S567L and C712R, which have previously been reported in a patient with bilateral tumors and in a family with low penetrance, respectively. Twelve of the mutations identified here have not been reported to date. These include a novel missense mutation, L662P, which was identified in two bilaterally affected siblings and their mother with unilateral retinoma. PMID- 10671069 TI - Proceedings from the XI Conference of Polish Association of Neuropathologists, Part I: Tumors of the Nervous System. Warszawa, May 13-15, 1999. PMID- 10671070 TI - The efficacy and effectiveness of a primary preventive dental programme in non fluoridated areas of Victoria, Australia. PMID- 10671071 TI - Survival after cardiac arrest outside hospital. PMID- 10671072 TI - Exercise four hour redistribution thallium-201 SPECT and exercise induced ST segment elevation in detecting viable myocardium in patients with acute MI. PMID- 10671073 TI - Quality of life four years after myocardial infarction: short form 36 scores compared with a normal population. PMID- 10671074 TI - Exercise testing, symptoms, and clinical outcome in aortic stenosis. PMID- 10671075 TI - Value of echocardiography in predicting long term outcome after heart transplantation. PMID- 10671076 TI - Good outcomes from cardiac surgery in the over 70s. PMID- 10671077 TI - Seeing what Alzheimer saw--with magnetic resonance microscopy. PMID- 10671078 TI - Statins: lower lipids and better bones? PMID- 10671079 TI - China's first international HIV vaccine meeting: the war of the roses. PMID- 10671080 TI - Embedded corneal foreign body in a child? Try a chalazion currette. PMID- 10671081 TI - Cytomegalovirus retinitis in AIDS patients: 1999 update. PMID- 10671082 TI - Bioengineering vs mechanical engineering for cardiovascular surgery in the next century. PMID- 10671084 TI - Report of the 25th Annual Meeting of the European Strabismological Association. Jerusalem, Israel, September 26-29, 1999. PMID- 10671083 TI - Venous thrombosis in traumatic brain injury. PMID- 10671085 TI - Report of the Annual Meeting of the Gunter K. von Noorden Visiting Professorship in Ophthalmology. Houston, Texas, September 16-17, 1999. PMID- 10671086 TI - Report of the Annual Meeting of the Texas Society for Pediatric Ophthalmology. Houston, Texas, September 18, 1999. PMID- 10671087 TI - Asp 280 residue is important in the activity of the Escherichia coli leader peptidase. PMID- 10671088 TI - Embryonic Stem Cells as Developmental Model in vitro. Preface. PMID- 10671090 TI - Planimetric models to evaluate the pennate muscle force length characteristics. PMID- 10671089 TI - Conjugate vaccines. PMID- 10671091 TI - Planimetric models to evaluate the pennate muscle force length characteristics. PMID- 10671092 TI - The worst of both worlds: poverty and politics in the Balkans. PMID- 10671093 TI - Defending JCAHO standards. PMID- 10671094 TI - Recent advances in genetic epidemiology. Special issue in honor of Professor Newton E. Morton's 70th birthday. PMID- 10671095 TI - Eighth International Congress of Pediatric Dermatology, Paris, France, 20 May 1998. PMID- 10671096 TI - Alopecia areata totalis/universalis and systemic corticosteroids. PMID- 10671097 TI - Pemphigus vulgaris associated with systemic lupus erythematosus. PMID- 10671098 TI - Tinea versicolor mimicking pityriasis rotunda. PMID- 10671099 TI - Cutaneous leishmaniasis in Aydin, Turkey. PMID- 10671100 TI - Lichenoid drug eruption due to cyanamide. PMID- 10671101 TI - Aplasia cutis congenita of the scalp associated with meningoencephalocele and situs inversus. PMID- 10671102 TI - Liesegang rings in a giant proliferating trichilemmal tumor. PMID- 10671103 TI - Phenelzine as adjuvant treatment for Behcet's disease? PMID- 10671104 TI - Cutaneous side-effects caused by Tegafur. PMID- 10671105 TI - Mycosis fungoides; evolution towards large-cell lymphoma. PMID- 10671106 TI - Importance of OMT cannot be overemphasized. PMID- 10671107 TI - Take the time to teach to ensure the legacy and future of our profession. PMID- 10671108 TI - Welcome to our future. PMID- 10671109 TI - Dietitians face the challenge of food allergies. PMID- 10671110 TI - Staying on track: the benefits of exercise combined with healthful eating. PMID- 10671111 TI - Role of closed mitral commissurotomy for mitral restenosis. AB - Out of 1184 consecutive cases of rheumatic mitral stenosis treated surgically by closed mitral commissurotomy (CMC) at NRS Medical College and Hospital, Calcutta, 20 (1.68%) were mitral valve restenosis. Twelve cases (60%) were females, The median age was 32 years. Duration between the first operation and reappearance of symptoms varied with a mean of 8 years. The previous operations were digital dilatation and instrumental dilatation in 6 and 14 cases respectively. History of thromboembolism was present in 4 cases. On echocardiography, calcification of the mitral valve was present in 2 cases, left atrial clot in 4 cases, associated mild to moderate mitral regurgitation in 6 cases and mild aortic regurgitation in 4 cases. All cases presented with New York Heart Association (NYHA) III and IV symptoms. Critical stenosis (mitral valve orifice less than 0.5 cm2) was present in 12 cases. Re-do CMC was undertaken in all cases with Tubb's dilator. Median operating time was 2.5 hours. Satisfactory split was achieved in 13 cases. One patient died during surgery. Four cases having less than satisfactory split were asymptomatic on follow-up. In one case no split was possible and in another, gross mitral regurgitation was noted postoperatively. These 2 cases had to undergo open heart surgery. It is concluded that re-do CMC is a feasible and suitable alternative in mitral restenosis even in the presence of complications. PMID- 10671112 TI - Proceedings of the 8th International Symposium on Platinum and Other Metal Coordination Compounds in Cancer Chemotherapy. Oxford, United Kingdom, 28-31 March 1999. PMID- 10671113 TI - Postictal psychosis related regional cerebral hyperfusion. PMID- 10671114 TI - Oncofetal matrix glycoproteins in cerebral arteriovenous malformations and neighbouring vessels. PMID- 10671115 TI - Hashimoto's encephalopathy presenting as "myxodematous madness". PMID- 10671116 TI - Alien hand sign in Creutzfeldt-Jakob disease. PMID- 10671117 TI - Recurrent peripheral neuropathy in a girl with celiac disease. PMID- 10671118 TI - Frontal release signs in older people with peripheral vascular disease. PMID- 10671119 TI - Factitious clock drawing adn constructional apraxia. PMID- 10671120 TI - Anosognosia and mania associated with right thalamic haemorrhage. PMID- 10671121 TI - Epileptic cardiac asystole. PMID- 10671122 TI - Respiratory insufficiency in a patient with hereditary neuropathy with liability to pressure palsy. PMID- 10671123 TI - Spinal accessory neuropathy and internal jugular thrombosis after carotid endarterectomy. PMID- 10671124 TI - Ischaemic stroke in a sportsman who consumed MaHuang extract and creatine monohydrate for body building. PMID- 10671125 TI - Petroclival meningioma as a cause of ipsilateral cervicofacial dyskinesis. PMID- 10671126 TI - Acute multifocal cerebral white matter lesions during transfer factor therapy. PMID- 10671127 TI - Fahr's disease and Asperger's syndrome in a patient with primary hypoparathyroidism. PMID- 10671128 TI - Hypertrophic atlantoaxial ligaments: an unusual cause of compression of the upper spinal cord. PMID- 10671129 TI - Selective hemihypaesthesia due to tentorial coup injury against dorsolateral midbrain: potential cause of sensory impairment after closed head injury. PMID- 10671130 TI - Early diagnosis of subependymal giant cell astrocytoma in children with tuberous sclerosis. PMID- 10671131 TI - Toluene induced postural tremor. PMID- 10671132 TI - Atypical form of amyotrophic lateral sclerosis: a new term to define a previously well known form of ALS. PMID- 10671133 TI - Isolated dysarthria. PMID- 10671134 TI - Motor cortical excitability in Huntington's disease. PMID- 10671135 TI - Critical closing pressure: a valid concept? PMID- 10671136 TI - High frequency stimulation of the subthalamic nucleus and levodopa induced dyskinesias in Parkinson's disease. PMID- 10671137 TI - Nitric oxide in acute ischaemic stroke. PMID- 10671138 TI - Historical note. Arnold Chiari, or "Cruvilhier Cleland Chiari" malformation. PMID- 10671139 TI - The consultation in art. PMID- 10671140 TI - More SANE program listings. PMID- 10671141 TI - [Antineoplastic effect of novel agents, UCN-01 on human tumor cells transplanted in nude mice]. PMID- 10671142 TI - Millennium bugs. AIDS in Africa threatens the United States. It should not act accordingly. PMID- 10671143 TI - Internet fervour hits genomics. PMID- 10671144 TI - Lobbyists anticipate more money for all. PMID- 10671145 TI - Should private enterprise take over. PMID- 10671146 TI - [Is it possible to spare eyebrows while undergoing radiotherapy?]. PMID- 10671147 TI - [Renal tubular acidosis and deafness due to mutation in the gene encoding the B1 subunit of H+ATPase]. PMID- 10671148 TI - [Role and physiologic importance of megalin in the internalization of vitamin D by endocytosis at the luminal pole of the proximal convoluted tubule]. PMID- 10671149 TI - [Does angiotensin II increase or decrease prostaglandin synthesis ?]. PMID- 10671150 TI - [ERA-EDTA Symposium on Renal Ischemia. Rimini, 6-9 June 1998]. PMID- 10671151 TI - Costs of otitis media? PMID- 10671152 TI - Evaluation of vaginal infections in adolescent women: can it be done without a speculum? PMID- 10671153 TI - Cancer in children: is the sample biased. PMID- 10671154 TI - Antibiotic use and parental home otoscopy. PMID- 10671155 TI - Health care for children in Afghanistan. PMID- 10671156 TI - Agricultural research. Windfall breeds fresh but vulnerable crop of grants. PMID- 10671157 TI - E.U. grabs food safety by the horns. PMID- 10671158 TI - Embryos attacked by mom's natural defenses. PMID- 10671159 TI - How rotavirus causes diarrhea. PMID- 10671160 TI - Reaping the plant gene harvest. PMID- 10671162 TI - Taxonomy. Researchers cash in on personalized species names. PMID- 10671161 TI - Meeting. Society for Integrative and Comparative Biology. Integrating the many aspects of biology. PMID- 10671163 TI - Science in response to basic human needs. PMID- 10671164 TI - Childhood cancer. PMID- 10671165 TI - Misallocation of CDC funds. PMID- 10671166 TI - Societal responsibilities. PMID- 10671167 TI - Societal responsibilities. PMID- 10671168 TI - Societal responsibilities. PMID- 10671169 TI - Societal responsibilities. PMID- 10671170 TI - Considering manure and carbon sequestration. PMID- 10671171 TI - Policy forum: genetic technologies. Monitoring and labeling for genetically modified products. PMID- 10671172 TI - Perspectives: plant biology. Some like it hot. PMID- 10671173 TI - Perspectives: transcription. A tail of histone acetylation and DNA recombination. PMID- 10671174 TI - Perspectives: protein synthesis. Unraveling the riddle of ProCys tRNA synthetase. PMID- 10671175 TI - Techview: molecular biology. Bead-based fiber-optic arrays. PMID- 10671176 TI - Discovery of the antidepressant and anti-emetic efficacy of substance P receptor (NK1) antagonists. AB - The development of small-molecule antagonists of the substance P (SP)-preferring tachykinin NK1 receptor during the past decade represents an important opportunity to exploit these molecules as novel therapeutic agents. On the basis of its anatomical localization and function, SP has been implicated in diverse pathophysiologies; of these, diseases of the CNS have been examined in the greatest detail. Although SP is best known as a pain neurotransmitter, it also controls vomiting and various behavioural, neurochemical and cardiovascular responses to stress. Recent clinical trials have confirmed the efficacy of NK1 receptor antagonists to alleviate depression and emesis but, surprisingly, not pain. Thus, multiple clinical trials, targeted to appropriate patient populations, are necessary to define the therapeutic potential of novel neurotransmitter ligands. PMID- 10671177 TI - [Torsion of the small intestine caused by gangrenous Meckel's diverticulum]. PMID- 10671178 TI - [The 75th anniversary of the St. Petersburg Scientific Society of Physiotherapists]. PMID- 10671179 TI - [Review of the materials of the 7th European Conference on Nutrition]. PMID- 10671180 TI - What topics will Acta cover in the new millenium? PMID- 10671181 TI - Microwave emissions from police radar. AB - This study evaluated police officers' exposures to microwaves emitted by traffic radar units. Exposure measurements were taken at approximated ocular and testicular levels of officers seated in patrol vehicles. Comparisons were made of the radar manufacturers' published maximum power density specifications and actual measured power densities taken at the antenna faces of those units. Four speed-enforcement agencies and one transportation research institute provided 54 radar units for evaluation; 17 different models, encompassing 4 frequency bands and 3 antenna configurations, were included. Four of the 986 measurements taken exceeded the 5 mW/cm2 limit accepted by the International Radiation Protection Association and the National Council on Radiation Protection and Measurement, though none exceeded the American Conference of Governmental Industrial Hygienists, American National Standards Institute, Institute of Electrical and Electronic Engineers, or Occupational Safety and Health Administration standard of 10 mW/cm2. The four high measurements were maximum power density readings taken directly in front of the radar. Of the 812 measurements taken at the officers' seated ocular and testicular positions, none exceeded 0.04 mW/cm2; the highest of these (0.034 mW/cm2) was less than 1% of the most conservative current safety standards. High exposures in the limited region directly in front of the radar aperture are easily avoided with proper training. Results of this study indicate that police officer exposure to microwave radiation is apparently minimal. However, because of uncertainty in the medical and scientific communities concerning nonionizing radiation, it is recommended that law enforcement agencies implement a policy of prudent avoidance, including purchasing units with the lowest published maximum power densities, purchasing dash/rear deck-mounted units with antennae mounted outside the patrol vehicle, and training police officers to use the "stand-by" mode when not actually using radar. PMID- 10671182 TI - Highly Unsaturated Fatty Acids in Nutrition and Disease Prevention. Proceedings of an international conference. Barcelona, Spain, November 4-6, 1996. PMID- 10671183 TI - Critical issues in academic orthodontics. PMID- 10671184 TI - More on indirect bonding. PMID- 10671185 TI - [Italian Bone Marrow Donor Registry]. PMID- 10671186 TI - Reductive dechlorination of tetrachloroethene to ethene by a two-component enzyme pathway. AB - Two membrane-bound, reductive dehalogenases that constitute a novel pathway for complete dechlorination of tetrachloroethene (perchloroethylene [PCE]) to ethene were partially purified from an anaerobic microbial enrichment culture containing Dehalococcoides ethenogenes 195. When titanium (III) citrate and methyl viologen were used as reductants, PCE-reductive dehalogenase (PCE-RDase) (51 kDa) dechlorinated PCE to trichloroethene (TCE) at a rate of 20 micromol/min/mg of protein. TCE-reductive dehalogenase (TCE-RDase) (61 kDa) dechlorinated TCE to ethene. TCE, cis-1,2-dichloroethene, and 1,1-dichloroethene were dechlorinated at similar rates, 8 to 12 micromol/min/mg of protein. Vinyl chloride and trans-1,2 dichloroethene were degraded at rates which were approximately 2 orders of magnitude lower. The light-reversible inhibition of TCE-RDase by iodopropane and the light-reversible inhibition of PCE-RDase by iodoethane suggest that both of these dehalogenases contain Co(I) corrinoid cofactors. Isolation and characterization of these novel bacterial enzymes provided further insight into the catalytic mechanisms of biological reductive dehalogenation. PMID- 10671187 TI - Physical training improves flow-mediated dilation in patients with the polymetabolic syndrome. AB - Endothelial dysfunction that can be detected as impaired flow-mediated dilation by ultrasonography is an early event in atherogenesis and has been demonstrated in healthy subjects with risk factors for atherosclerosis many years before the appearance of atheromatous plaques. We examined the influence of physical training on flow-mediated dilation in patients with the polymetabolic syndrome. Twenty-nine asymptomatic men aged 40 to 60 years with the polymetabolic syndrome were randomly divided between the control group and the training group, which trained 3 times a week for 12 weeks. On high-resolution ultrasound images, the diameter of the brachial artery was measured at rest, after reactive hyperemia (causing flow-mediated, endothelium-dependent dilation), and after sublingual glyceryltrinitrate (causing endothelium-independent vasodilation) in all subjects before and after the training period. The training program induced an increase of 18% in physical fitness. Flow-mediated dilation increased from 5.3+/-2.8% to 7.3+/-2.7% (P<0. 05). There was no change in body mass index, blood pressure, insulin resistance, lipids, and big endothelin-1 in either group. Flow-mediated dilation measured before training was negatively correlated with resting heart rate, waist-to-hip ratio, and insulin resistance. Resting heart rate emerged as the only independent determinant, which explained 22% of the variation in flow mediated dilation. In conclusion, our findings suggest that a 3-month physical training program, which improved maximal exercise capacity, enhances flow mediated dilation in patients with the polymetabolic syndrome. PMID- 10671188 TI - Glucocorticoid production in the murine thymus. AB - Glucocorticoid hormones are known to act as important modulatory factors in the development of autoimmune diseases, and to play an important role in thymic T cell selection. There seems to be a finely balanced equilibrium between the apoptosis-inducing effects of glucocorticoid and T cell receptor ligand binding. Here we are investigating whether glucocorticoid-induced T cell apoptosis is mainly dependent on circulating glucocorticoid levels or if the thymus itself is able to produce glucocorticoids. To this end, we attempted to demonstrate enzyme activities of the whole set of steroidogenic enzymes for the synthesis of glucocorticoids in murine thymic tissue. We isolated steroidogenic organelles from thymic tissue, incubated these with radioactive (precursor) steroids in vitro, and visualized the resulting products by thin-layer chromatography. Our results show that the thymus possesses all enzymes and cofactors required for glucocorticoid production. However, an intact thymic architecture is necessary for glucocorticoid production, since 11beta-hydroxylase was not detected in irradiated thymi or in a thymic epithelial cell line. The results of these experiments show that the whole glucocorticoid metabolism takes place within the thymus. This finding provides the biochemical basis for the in situ effects of glucocorticoid hormones on thymocyte development and selection. PMID- 10671189 TI - Endogenous IL-12 synthesis is not required to prevent hyperexpression of type 2 cytokine and antibody responses. AB - Endogenous IL-12 production is hypothesized to play an essential role preventing spontaneous expression of type 2 responses, acting as a natural inhibitor limiting development of immediate hypersensitivity. Here, IL-12-deficient p35(- / -) and p40(- / -) mice were used to examine the role of endogenous IL-12 and p40 homodimer during in vivo development of exogenous antigen-driven responses. In the absence of deliberate immunization, IL-12-deficient mice exhibited greatly reduced serum IgG2a but IgG1 / IgE levels no higher than controls. Immunization to elicit polarized ovalbumin-specific type 1 or type 2 dominant responses, or using Trichinella spiralis extract in the absence of adjuvants, led to IFN-gamma production of approximately 10 % of C57BL / 6 controls yet the kinetics and intensity of primary and secondary type 2 cytokine (IL-4, IL-5, IL-13) and antibody (IgG1, IgE) responses, as well as functional IL-12 receptor expression, were consistently unaltered. Thus, while IL-12 provides an important positive signal for Th1 development, antigen exposure in its absence does not lead to generalized enhancement of type 2 cytokine or antibody responses. The data argue that endogenous IL-12 production is not required as a constitutive negative regulator limiting induction or expression of type 2 effector responses. PMID- 10671190 TI - Genetic dissection of B cell traits in New Zealand black mice. The expanded population of B cells expressing up-regulated costimulatory molecules shows linkage to Nba2. AB - B cell abnormalities are a prominent feature of the immunologic derangement in NZB and NZB / W mice. We recently demonstrated that these mice have an increased proportion of splenic B cells expressing B7.1 and elevated levels of B7.2 and ICAM-1 that possess the characteristics of marginal zone B cells (CD23(low / -) CD5(-) CD44(hi) CD24(hi) IgD(- / low) IgM(hi)) and are found as early as 4 - 6 weeks of age. These findings suggest that activated B cells in NZB and NZB / W mice could serve a costimulatory function leading to activation of autoreactive T cells. However, it remains unclear whether there is any association between B abnormalities and nephritis in these mice. Here we have used genetic mapping techniques to address this issue. We show that increases in the proportion of B cells expressing costimulatory molecules, serum IgM levels, the number of IgM ELISpots, and IgG anti-single-stranded (ss) DNA antibody production, are significantly associated with a chromosomal region that overlaps with Nba2, a genetic locus previously linked to nephritis. Based on these findings we propose that immune mechanisms leading to polyclonal B cell activation and up-regulation of costimulatory molecules in these mice play a central role in the loss of tolerance that leads to production of pathogenic autoantibodies. PMID- 10671191 TI - CD3 activation induces concentrative nucleoside transport in human T lymphocytes. AB - Nucleoside transport, assessed by measuring deoxythymidine influx, was investigated in normal and CD3-activated human peripheral blood mononuclear cells (PBMC) and in the CEM cell line. On both cell types, an equilibrative nitrobenzylmercaptopurine (NBMPR)-sensitive (es) transporter encoded by the hENT(1) gene was identified on resting cells, although the expression level was about 20-fold higher on CEM cells than on resting peripheral T lymphocytes. After stimulation with anti-CD3, a strong increase of nucleoside transport was observed in PBMC accompanied by a mild augmentation of NBMPR binding sites on the cell surface. Most of this improved transport capacity was NBMPR insensitive, dependent on Na(+) concentration in the medium, and displayed the features of a concentrative process. Similar results were obtained with CEM cells despite their high basal es level, indicating that the induction of a concentrative process for nucleoside salvage is a specific metabolic response associated with antigen driven stimulation. In CEM cells, this induction did not affect the growth rate. The concentrative transporter involved does not correspond to any of those which have been cloned so far. Molecular characterization of this transporter should provide a new marker of antigen stimulation and will allow to define whether activation of the corresponding gene is under the control of TCR-CD3-induced second messengers. PMID- 10671192 TI - Different doses of agonistic ligand drive the maturation of functional CD4 and CD8 T cells from immature precursors. AB - MHC molecules are normally required for the development of thymocytes from the CD4(+)CD8(+) double-positive to the CD4 or CD8 single-positive stage. Here we show that mitogenic plant lectins can substitute for MHC molecules in driving the differentiation of phenotypically and functionally mature CD4 as well as CD8 T cells. Interestingly, lectin dosage determines whether CD4 or CD8 cells are generated, indicating that variation of cumulative signal strength (not necessarily signal quality) can result in an apparent switching of lineage preference. Thymocyte perception of differentiation-inducing signals is modulated by the cellular context, since stimuli that yield CD8 cells in the context of the thymic microenvironment fail to do so in suspension culture and generate CD4 progeny instead. Finally, we show that lectin-generated single-positive thymocytes retain the ability to respond to the ligands initially used to drive their differentiation. Our results call into question generalizations and predictions made from other experimental systems and reveal that thymocyte selection is considerably more flexible than had been anticipated. PMID- 10671193 TI - IL-10 deficiency prevents IL-5 overproduction and eosinophilic inflammation in a murine model of asthma-like reaction. AB - Eosinophilic inflammation and bronchial mucus secretion are among the characteristic pathological changes in asthmatic reaction, which is mediated by Th2 type responses. Although it belongs to Th2 cytokines especially in the mouse, IL-10 is often considered an inhibitory cytokine for both Th1 and Th2 cells. In the present study, using a murine asthma model induced by ovalbumin (OVA), we demonstrated that endogenous IL-10 is critical for the development of asthma-like responses. Specifically, in comparison with wild-type controls, IL-10 gene knockout (KO) mice showed significantly reduced IL-5 production, eosinophilic inflammation and mucus production without notable changes in IL-4 and IgE responses following i. p. sensitization and subsequent intranasal challenge with OVA. In addition, Th1-related cytokine (IFN-gamma and IL-12) production in IL-10 KO mice was significantly higher than that in wild-type mice. The results suggest that endogenous IL-10 plays an important role in promoting pulmonary eosinophilic inflammatory reaction and mucus production during asthmatic reaction. The data also argue that IL-10 may be more influential in the development of IL-5 producing Th2 cells which differ from typical Th2 cells producing both IL-4 and IL-5. PMID- 10671194 TI - Co-stimulation of antigen-specific CD4 T cells by 4-1BB ligand. AB - 4-1BB is a member of the TNF receptor family predominantly expressed on activated T cells, and binds an inducible ligand found on B cells, macrophages and dendritic cells. Whereas ligation of 4-1BB has been shown to enhance response of purified CD8 T cells to mitogens, and to augment NK activity and generation of cytotoxic T lymphocytes in vivo, there are little direct data on 4-1BB action during CD4 responses. Using pigeon cytochrome c-presenting fibroblast antigen presenting cells transfected with 4-1BB ligand (4-1BBL), we show that engaging 4 1BB on naive CD4 cells promotes proliferation, cell cycle progression and IL-2 secretion, and suppresses cell death, all to a similar extent as B7-1 engagement of CD28. In addition, 4-1BBL synergizes with B7 and ICAM to enhance naive CD4 proliferation when antigen is limiting. 4-1BBL alone, and to a greater extent with B7, also augmented IL-2 secretion resting antigen-experienced CD4 cells, as typified by T helper clones, whereas short-term effector cells showed similar levels of proliferation and cytokine secretion regardless of whether 4-1BB was engaged. A major role in augmenting IFN-gamma, IL-4 or IL-5 was not demonstrated. Blocking studies with activated B cells presenting antigen showed that 4-1BB participates in promoting IL-2 production by resting CD4 cells, confirming that 4 1BBL can play a role in antigen-specific CD4 T cell responses. PMID- 10671195 TI - Down-regulation of antigen-specific antibody production by TCR antagonist peptides in vivo. AB - The efficacy of TCR antagonist peptides in inhibition of antigen-specific antibody production and T cell responses in vivo was evaluated. Among amino acid substituted analogs of a peptide corresponding to residues 119 - 133 of bovine beta-lactoglobulin (p119 - 133), pR124Q and pD129S, prepared by substitution of Gln and Ser for Arg(124) and Asp(129), respectively, have been shown to display TCR antagonist activity for three out of four distinct p119 - 133-specific T cell clones and for polyclonal T cells derived from p119 - 133-immunized C57BL / 6 mice. Both pD129S and pR124Q inhibited in vivo priming and subsequent activation of T cells by p119 - 133 when co-injected with p119 - 133 into mice, as shown by the decreased proliferation of T cells in response to p119-133 in vitro. pD129S significantly inhibited production of anti-p119 - 113 antibodies of IgG1, IgG2b and IgE isotype in vivo when co-injected into mice together with p119 - 133 at the time of the first immunization. However, pR124Q was totally ineffective in inhibition of the antibody responses. Anti-p119 - 133 antibodies from p119 - 133 immunized mice could bind to pR124Q but not to pD129S, suggesting that the difference in cross-reactivity is responsible for the different effect of these two peptides on specific antibody production. Our findings demonstrate that a single TCR antagonist peptide can inhibit antigen-specific polyclonal antibody production when this antagonist peptide does not cross-react with the antibody elicited in response to an antigenic peptide. PMID- 10671196 TI - Direct anti-inflammatory effect of a bacterial virulence factor: IL-10-dependent suppression of IL-12 production by filamentous hemagglutinin from Bordetella pertussis. AB - IL-12 plays a critical role in protective immunity against intracellular pathogens by promoting the development of Th1 cells. Here we demonstrate that filamentous hemagglutinin (FHA), a virulence factor of Bordetella pertussis, is capable of suppressing IL-12 production by macrophages. FHA inhibited IL-12 secretion by a macrophage cell line or ex vivo alveolar macrophages in response to Escherichia coli or B. pertussis lipopolysaccharide (LPS) and IFN-gamma. Antibodies to FHA or denaturation of FHA abrogated the inhibitory effect. Injection of mice with FHA suppressed IL-12 and IFN-gamma levels in the serum in response to i. v. injection of LPS in a model of septic shock. The suppressive effect of FHA was specific for IL-12, since the production of TNF-alpha, IL-6 and IL-10 was not suppressed, and production of IL-6 and IL-10 was up-regulated. Antibody blocking studies revealed that the inhibitory effect of FHA on IL-12 production was dependent on IL-10. Since FHA is secreted at high levels and local T cell responses are suppressed during B. pertussis infection, the findings suggest that FHA may be a critical virulence factor in facilitating pathogen persistence in the respiratory tract by suppressing or delaying the development of cell-mediated immunity. PMID- 10671197 TI - A subunit vaccine candidate region of the Entamoeba histolytica galactose adherence lectin promotes interleukin-12 gene transcription and protein production in human macrophages. AB - The cysteine-rich region of the 170-kDa subunit galactose-adherence lectin (Gal lectin) of Entamoeba histolytica is a subunit vaccine candidate and a protective antigen in the gerbil model of amebiasis. Macrophage-mediated immunity is important for protection against E. histolytica and is activated by Th1 cytokines. As Th1 differentiation is promoted by IL-12, we investigated what portion of the Gal-lectin could stimulate IL-12 in human THP-1 macrophages. Native Gal-lactin stimulated IL-12 p40 / p35 mRNA expression in a dose- and time dependent manner as measured by reverse transcriptase-PCR. Human immune serum and Gal-lectin mAb inhibition studies identified amino acids (aa) 596 - 998 as immunogenic and containing the IL-12 inducing domain. IFN-gamma priming augmented Gal-lectin-induced IL-12 mRNA expression independent of TNF-alpha and IL-1beta, and was required for IL-12 p70 protein production from macrophages and human peripheral blood mononuclear cells. Gal-lectin plus IFN-gamma stimulated IL-12 p40 and p35 gene transcription with stable mRNA transcripts and a differential requirement for protein synthesis. These results suggest that aa 596 - 998 of the Gal-lectin can confer Th1-mediated protection against amebiasis through IL-12 induction. PMID- 10671198 TI - Pristane-induced arthritis in mice selected for maximal or minimal acute inflammatory reaction. AB - The role of inflammatory and specific immune responses in pristane-induced arthritis (PIA) was investigated in mouse lines produced by bi-directional selective breedings for maximal (AIRmax) or minimal (AIRmin) acute inflammatory reaction, comparing the outcome of PIA and the humoral and cellular response to hsp65. Symptoms of arthritis were detected in 50 % AIRmax mice 120 days after pristane injection, reaching a maximal incidence of 65 %, whereas only 7 % of AIRmin mice developed arthritis within an observation period of 200 days. The production of IgG antibody against hsp65 was found to be similar on both lines, although the IgG1 isotype was predominant in AIRmax, and IgG2a in AIRmin line. In vitro T cell proliferation to hsp65 was similar in the two lines, however, ELISPOT assays carried out soon after pristane treatment, demonstrated higher numbers of IL-6-, TNF-alpha- and IL-4-secreting cells in the spleen of AIRmax than in AIRmin mice, while higher numbers of IFN-gamma-producing cells were found in AIRmin mice. These results suggest a major participation of acute inflammatory mechanisms in the susceptibility to PIA. The genetic background which determines high or low AIR favors a Th2-like response in susceptible AIRmax and Th1-like response in resistant AIRmin mice at the initial phase of arthritis induction. PMID- 10671199 TI - Dichotomic effects of IFN-gamma on the development of systemic lupus erythematosus-like syndrome in MRL-lpr / lpr mice. AB - Systemic lupus erythematosus (SLE)-prone female MRL-lpr / lpr (MRL-lpr) mice were treated with mouse or rat IFN-gamma under different experimental conditions, both prophylactically in 6- to 8 week-old animals and therapeutically in 12- to 18 week-old SLE-affected mice. It was found that IFN-gamma heterogeneously modulated the course of the disease in MRL-lpr mice. When administered prophylactically, IFN-gamma favorably modulated the histological, serological and clinical signs of the disease. Relative to untreated or PBS-treated control animals, the MRL-lpr mice which received IFN gamma were virtually free of inflammatory infiltration of the kidneys and the lungs, had lower levels of azotemia with reduction of both circulating IgG1, IgG2a and IgG3 and anti-double strand (ds) and single strand (ss) DNA antibodies, milder skin vasculitis, significantly reduced enlargement of their lymph nodes and lower weight of the spleens. IFN-gamma also lowered the rate of mortality of MRL-lpr mice. In contrast to these findings, therapeutically administered IFN-gamma worsened the course of the disease in MRL-lpr mice, which exhibited increased proteinuria, higher levels of IgG2a and IgG3 and anti-ds and ss DNA antibodies, more aggressive nephritis and died at an earlier age than PBS treated control mice. The dichotomic effect of IFN-gamma on disease manifestation in MRL-lpr mice offers new insights into the complex role of this cytokine in the regulation of systemic autoimmunity such as SLE. PMID- 10671200 TI - Multiple cross-reactive self-ligands for Borrelia burgdorferi-specific HLA-DR4 restricted T cells. AB - T cell recognition of self antigens is a key event in the pathogenesis of autoimmune diseases. To date, the initial events that trigger autoreactive T cells are unknown. The "molecular mimicry" hypothesis predicts that during an infection T cells that recognize both a microbial antigen and a related self peptide become activated and cause autoimmune disease. We have systematically examined the recognition of self antigens by HLA-DR4-restricted T cells specific for peptides of the outer surface protein A (OspA) of Borrelia burgdorferi, the etiological agent of Lyme disease. We used the peptide spot synthesis technique for complete peptide substitution analyses of two immunodominant OspA epitopes. Each amino acid residue of the epitopes was substituted with all 20 naturally occurring amino acids and the altered peptides were tested for recognition by a panel of OspA-specific T cells. The binding motifs (supertopes) revealed by these analyses were used to screen public databases for matching human or murine peptides. Several hundred peptides were identified by this search and synthesized. Of these, 28 were recognized by OspA-specific T cells. Thus, T cell cross-reactivity is a common phenomenon and the existence of cross-reactive epitopes alone does not imply molecular mimicry-mediated pathology and autoimmunity. PMID- 10671201 TI - The BOB.1 / OBF.1 co-activator is essential for octamer-dependent transcription in B cells. AB - The BOB.1 / OBF.1 / OCA-B protein (henceforth designated as BOB.1 / OBF.1) is a B cell-specific co-activator of the Oct1 and Oct2 transcription factors. It is involved in mediating the transcriptional activity of these proteins. Surprisingly, animals deficient for BOB.1 / OBF.1 showed normal expression of genes that contain an octamer motif in their regulatory regions. Here we have addressed the role of BOB.1 / OBF.1 for octamer-dependent transcription. We show that promoters exclusively dependent on functional octamer motifs are completely inactive in BOB.1 / OBF. 1-deficient B cells. The lack of activity is a direct consequence of lack of the co-activator. To show this, a hormone-regulated conditional allele of BOB.1 / OBF.1 was introduced into the BOB.1 / OBF.1 deficient B cells. This resulted in the hormone-dependent transcriptional activity of octamer-dependent reporters in these cells. The BOB.1 / OBF.1 requirement for octamer promoter function was also observed when an authentic immunoglobulin kappa-promoter was assayed. BOB.1 / OBF.1 dependence could not be overcome by including the strong enhancer element from the immunoglobulin heavy chain gene. Induction of pre-B cells with lipopolysaccharide led to increased Oct2 levels but did not significantly increase octamer-dependent transcription in BOB.1 / OBF.1-deficient B cells. Thus, these results demonstrate that BOB.1 / OBF.1 itself is a non-redundant protein in B cells and absolutely required for octamer-dependent transcriptional activity. PMID- 10671202 TI - Elevated type 1, diminished type 2 cytokines and impaired antibody response are associated with hepatotoxicity and mortalities during Schistosoma mansoni infection of CD4-depleted mice. AB - During murine Schistosoma mansoni infections parasite eggs evoke a type 2 cytokine-dependent and CD4(+) T cell-mediated granulomatous response in the liver. In this study CD4(+) T cell-depleted CBA / Ca mice developed hepatic steatosis and had high mortalities during early acute schistosome infection. CD4 depleted mice had smaller liver granulomas and reduced hepatic fibrosis. The hepatocytotoxicity was characterized by microvesicular steatosis and neutrophil infiltration. The livers of depleted mice had similar levels of apoptosis as control infected mice but had a marked increase in lipid peroxidation indicative of their livers being under oxidative stress. CD4-depleted mice had impaired egg excretion and exacerbated intestinal pathology. A type 1 cytokine-dominated response was present in infected CD4-depleted mice and relatively reduced production of type 2 cytokines. Antibody responses to parasite antigens were also substantially reduced. Transfer of immune serum or IgG significantly delayed mortalities in depleted mice and prevented hepatocyte damage. Although biasing the cytokine dichotomy to a type 1-dominated response during murine schistosome infection is desirable with respect to certain pathological processes, i. e. it will reduce the granulomatous inflammation and hepatic fibrosis, these effects contribute to fatal pathology if there is reduced protective type 2 cytokines and a defect in antibody responses. PMID- 10671203 TI - Mouse FcgammaRII is a negative regulator of FcgammaRIII in IgG immune complex triggered inflammation but not in autoantibody-induced hemolysis. AB - Murine low-affinity receptors for IgG, FcgammaRII and FcgammaRIII, differ by their distinct capacities in mediating down-regulation or activation of cellular effector functions, respectively. In this study, antibodies detecting the mouse Ly-17.1 / 2 alloantigen system are demonstrated to be specific for FcgammaRII with no cross-reactivities to other FcgammaR, including FcgammaRIII. Using these FcgammaRII-specific monoclonal antibodies (mAb), the significance of FcgammaRII inhibition of FcgammaRIII was examined in two models of autoantibody [autoimmune hemolytic anemia (AIHA)]- and IgG immune complex-induced (Arthus reaction) inflammation in C57BL / 6 mice in comparison with FcgammaRII(- / -) and FcgammaRIII(- / -) mice. Our results demonstrate that both FcgammaRIII and FcgammaRII contributed to the binding of erythrocytes opsonized with the pathogenic IgG1 autoreactive anti-murine red blood cell antibody 105-2H. However, the functional blocking with anti-FcgammaRII mAb in C57BL / 6 mice and the lack of FcgammaRII expression in FcgammaRII(- / -) mice, which both lowered the threshold level of FcgammaRIII-triggered phagocytosis in vitro, did not results in enhanced disease development of 105-2H mAb-induced AIHA in vivo. This was in sharp contrast to cutaneous Arthus reaction, where FcgammaRIII-mediated activation was inhibited by FcgammaRII. Together these results show that murine AIHA is markedly different from other FcgammaR-dependent inflammatory diseases where FcgammaRIII is normally counterregulated by FcgammaRII. PMID- 10671204 TI - Human afferent lymph from normal skin contains an increased number of mainly memory / effector CD4(+) T cells expressing activation, adhesion and co stimulatory molecules. AB - We investigated the T cell population in the afferent lymph and the peripheral blood with regard to expression of activation, adhesion and co-stimulatory molecules and cytokine profile by immunohistochemistry and flow cytometry. The majority of the lymphoid cell population in the afferent lymph were CD4(+), CD45RO(+) T cells expressing the alpha beta TCR. An increased percentage of the T cells expressed activation molecules like HLA-DR, CD25, CD26, CD69 as well as adhesion and co-stimulatory molecules like CD54, CD154 / 40 ligand. Furthermore, T cells in the afferent lymph predominantly expressed the type 1 cytokine IFN gamma, whereas type 2 cytokines such as IL-4, IL-5, IL-10 were not or barely detectable. Interestingly, dendritic cells expressing IL-12 were also found in close association with some lymphocytes, indicating that these contacts may be important in promoting Th 1 cells. In conclusion, an increased number of mainly CD4(+) memory / effector T cells, expressing activation, adhesion and co stimulatory molecules migrate through the afferent skin-derived lymph in humans. Furthermore, our data demonstrate the dominance of a type 1 cytokine profile in these T cells and suggest that they have an important function in the immune surveillance against pathogens or other antigens in the skin and its associated lymphoid tissue. PMID- 10671205 TI - Inhibition of Th1 development and treatment of chronic-relapsing experimental allergic encephalomyelitis by a non-hypercalcemic analogue of 1,25 dihydroxyvitamin D(3). AB - 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] inhibits production of IL-12, a cytokine involved in the development of Th1 cells and in the pathogenesis of Th1 mediated autoimmune diseases. Here, we show that 1,25(OH)(2)D(3) and a non hypercalcemic analogue are selective and potent inhibitors of Th1 development in vitro and in vivo without inducing a deviation to the Th2 phenotype. Administration of 1,25(OH)(2)D(3) or its analogue prevents chronic-relapsing experimental allergic encephalomyelitis (CR-EAE) induced by the myelin oligodendrocyte glycoprotein (MOG) peptide 35 - 55 (MOG(35 - 55)) in Biozzi AB / H mice. The inhibition of EAE induction is associated with a profound reduction of MOG(35 - 55)-specific proliferation and Th1 cell development. Importantly, the non-hypercalcemic analogue also provides long-term protection from EAE relapses induced by immunization with spinal cord homogenate when administered for a short time at symptom onset or even after the first peak of disease. Neuropathological analysis shows a reduction of inflammatory infiltrates, demyelinated areas and axonal loss in brains and spinal cords of treated mice. These resuls indicate that inhibition of IL-12-dependent Th1 cell development is associated with effective treatment of CR-EAE and suggest the feasibility of an approach based on low molecular weight inhibitors of IL-12 production in the treatment of multiple sclerosis. PMID- 10671206 TI - A role for MHC class I down-regulation in NK cell lysis of herpes virus-infected cells. AB - NK cells represent an efficient first line of defense against virus infection, preceding the generation of adaptive T cell responses. However, the NK cell receptors involved in the recognition of virus-infected cells remain ill defined. We studied the in vitro response of isolated human NK cell clones to cells infected by the herpes viruses, herpes simplex virus (HSV) and human cytomegalovirus (HCMV). Both HSV and HCMV were found to induce NK cell cytotoxicity by down-regulating HLA-C molecules engaged in the triggering of killer inhibitory receptors (KIR). This conclusion was further substantiated by the finding that expression of viral genes known to interfere with MHC class I expression, such as the TAP inhibitor ICP47 of HSV and the MHC class I-destroying US11 protein of HCMV, was sufficient to trigger the cytotoxicity of NK cell clones expressing an inhibitory KIR for HLA-C. These results show for the first time that MHC class I down-regulation could render cells infected with herpes viruses susceptible to NK cell killing, thus demonstrating a role for KIR in the recognition of virally infected cells. PMID- 10671207 TI - Lactate dehydrogenase A-dependent surface expression of immature thymocyte antigen-1: an implication for a novel trafficking function of lactate dehydrogenase-A during T cell development. AB - A possible involvement of lactate dehydrogenase A (LDHA) in the translocation of a thymic differentiation antigen, immature thymocyte antigen-1 (IMT-1), from cytoplasm to cell surface membrane during thymocyte differentiation is described. Transfection of cDNA for LDHA, but not LDHB, into EL4 cells, which expressed IMT 1 in the cytoplasm but not on the cell surface, induced the expression of IMT-1 on the cell surface. This translocation process seemed to be dependent on the translation of LDHA cDNA in EL4 cells, as well as the native structure of LDHA composing of coenzyme binding domain, catalysis domain, and subunit contact domain. Immunoprecipitation analysis revealed that LDHA could be co-precipitated with IMT-1 from the cell surface of EL4 cells that had been transfected with LDHA cDNA, suggesting that some of LDHA is associated with cell surface IMT-1 on the transfectants. Flow cytometry analysis of thymocyte subpopulations showed that some thymocytes at the CD4(-)CD8(-) double negative stage express both IMT-1 and LDHA on their surface. These data indicate that LDHA, in addition to its function in the metabolism in the glycolytic pathway, may have a novel function in the expression of a cell surface antigen during T cell development. PMID- 10671208 TI - Differential effects on T cell and NK cell development by tissue-specific expression of H-2D(d) transgene. AB - The effect of tissue-specific expression of the MHC class I molecule H-2D(d) on T cell and NK cell specificity was studied in transgenic mice expressing the H 2D(d) gene under the control of the mouse metallothionein-I promoter. MTD mice expressed high amounts of H-2D(d) in the liver, intestine and testis, but only minute amounts in the thymus, spleen and kidney. Zinc administration resulted in a 1.5- and 8.5-fold increase in H-2D(d) expression in the liver and the intestine, respectively, but did not affect expression in the other organs tested. T cell tolerance developed towards H-2D(d) in MTD mice, even in the absence of zinc. In contrast, NK cell-mediated natural resistance against lymphoma grafts was not seen in MTD mice, despite zinc administration. NK cells in MTD mice also failed to develop self tolerance to H-2D(d). The lack of functional effects did not result from inability of NK cells in MTD mice to interact with H-2D(d), as down-regulation of Ly49A receptor expression was observed on liver NK cells in MTD mice. Our data reveal a difference between T cells and NK cells in their requirements for MHC class I molecules in specificity development. PMID- 10671209 TI - The human immunoglobulin loci introduced into mice: V (D) and J gene segment usage similar to that of adult humans. AB - Variable gene segments of the human immunoglobulin loci are represented in the human peripheral repertoire at different frequencies. XenoMouse strains contain approximately 2 megabases of the human immunoglobulin heavy and kappa light chain loci that functionally recapitulate the human humoral immune system. Analysis of human antibody transcripts from XenoMouse spleens and lymph nodes revealed that V, D and J gene segment utilization from these unimmunized animals were nearly identical to the gene segment utilization reported for humans with extensive antigenic histories. PMID- 10671210 TI - Acellular components of Chlamydia pneumoniae stimulate cytokine production in human blood mononuclear cells. AB - Accumulating evidence suggest that infection with Chlamydia pneumoniae is associated with atherosclerosis, but the mechanisms involved remain unclear. Inflammation is important in the initial phase of atherogenesis, and cytokines are important in the initiation and progression of inflammation. The aim of this study was to assess the capacity of acellular components of C. pneumoniae to stimulate the production of pro-inflammatory cytokines and chemokines. Peripheral blood mononuclear cells were stimulated in vitro with sonicated C. pneumoniae. Significant amounts of TNF-alpha, IL-1, IL-6, IL-8, monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1alpha (MIP-1alpha) were produced. Inhibition of endotoxin using polymyxin B revealed that chlamydial endotoxin plays a minor role in the cytokine induction. Neutralization of TNF by TNF-binding protein and blockade of IL-1 receptors by IL-1 receptor antagonist revealed that TNF, IL-1 and IL-6 production was independent from each other, whereas IL-8 synthesis was strongly dependent on endogenous TNF and IL-1. In contrast, synthesis of MCP-1 and MIP-1alpha was dependent on endogenous TNF, but not IL-1. In conclusion, acellular components of C. pneumoniae are a potent stimulus for cytokine production, and this mechanism may have an important role in the inflammatory aspects of atherogenesis. PMID- 10671211 TI - Differential expression of the B cell-restricted molecule CD22 on neonatal B lymphocytes depending upon antigen stimulation. AB - Newborns respond poorly to certain antigens and produce mainly IgM antibodies. By flow cytometry we analyzed on neonatal and adult B cells the expression of CD22, a B cell receptor (BCR)-associated membrane molecule, known as negative modulator of BCR signaling. After T cell-independent (TI-)stimulation with anti-mu F(ab')(2) fragments we found a dramatic decrease in the percentage of neonatal CD22(+) B cells and CD22 mean fluorescence intensity (MFI) shift, whereas adult B cells remained unaffected. Survival and proliferation rates of neonatal B cells were higher compared to adult B cells whereas the degrees of apoptosis and necrosis were comparable. Surprisingly, after stimulation with lower doses of anti-mu apoptosis as well as proliferation increased significantly in contrast to adult B cells. T cell-dependent (TD)-stimulation with anti-CD40 monoclonal antibody and IL-4 resulted in a dramatic increase in the percentage of CD22(+) neonatal B cells in contrast to unaffected adult B cells. CD22 MFI shifts showed no significant changes, respectively. The survival rate was higher for adult B cells, whereas apoptosis and cell death were comparable. These results suggest that TI antigens lower the neonatal BCR signaling threshold via down-regulation of CD22, resulting in hyperresponsive B cells apt to premature apoptosis. On the other hand, up-regulation of CD22 after TD stimulation may allow increased inhibiting influence of CD22 on neonatal BCR signaling, impairing B cell activation and differentiation. PMID- 10671212 TI - CXCR5-deficient mice develop functional germinal centers in the splenic T cell zone. AB - The chemokine receptor CXCR5 is thought to be essential for the migration of B cells into the network of follicular dendritic cells in the spleen. However, as shown here, B cells and follicular dendritic cells do co-localize, albeit aberrantly, even in the absence of CXCR5. In mice lacking CXCR5 both cell types are found in a broad ring around the sinuses of the marginal zones. Upon immunization with the T cell-dependent antigen 2-phenyl-oxazolone, ectopic germinal centers develop in the periarteriolar lymphocyte sheath. A network of follicular dendritic cells forms in the vicinity of the central arteriole within which the antigen-activated B cells proliferate. The analysis of the expressed V gene repertoire revealed that during B cell proliferation, hypermutation is activated and V region genes accumulate somatic mutations. The pattern of somatic mutations suggests that affinity selection may occur. This analysis confirms that in CXCR5-deficient mice, the organization of splenic primary follicles is severely impaired. However, within the T cell zone a micro-environment is built up, which provides all requirements needed for the affinity maturation to take place. PMID- 10671213 TI - Human KLRF1, a novel member of the killer cell lectin-like receptor gene family: molecular characterization, genomic structure, physical mapping to the NK gene complex and expression analysis. AB - The human NK gene complex localized on chromosome 12p12.3 - p13.2 codes for several lectin-like receptor genes expressed by NK cells as well as by other hematopoietic cells. In this study, by using the expressed sequence tag database we identified a novel receptor gene, designated as killer cell lectin-like receptor, subfamily F, member 1 (KLRF1), encoding a putative type II transmembrane glycoprotein. The KLRF1 gene has been localized on the high resolution physical map of chromosome 12p. The genomic structure of the KLRF1 gene and the existence of one spliced variant are also described. KLRF1 was expressed at the mRNA level in peripheral blood leukocytes, activated NK cells, monocytes and NK and myeloid cell lines. The presence of two immunoreceptor tyrosine-based inhibitory-like motifs within the cytoplasmic tail of KLRF1 suggests an inhibitory role in NK cell and monocyte activity. PMID- 10671214 TI - CD7 expression distinguishes subsets of CD4(+) T cells with distinct functional properties and ability to support replication of HIV-1. AB - Enrichment of a subset of CD4(+)CD45R0(+)CD7(-) T cells has been observed in HIV infected individuals. We have investigated the ability of CD7(+) and CD7(-) T cells to support replication of HIV and show that virus replicates preferentially in CD7(+) cells. Several possible mechanisms that may underlie such differences in susceptibility to HIV were studied. Our data demonstrate that mitogen stimulation induces poor expression of CD25 and IL-2 in CD7(-) compared with CD7(+) cells. We also show that uninfected CD7(-) cells are more resistant to mitogen-induced apoptosis than CD7(+) cells. Our data support the view that the CD7(-) subset is inherently resistant to HIV replication and that this is due in part to reduced CD25 expression and IL-2 production. PMID- 10671215 TI - HLA-G inhibits the transendothelial migration of human NK cells. AB - The expression of the non-classical MHC class I molecule HLA-G is normally restricted to the placenta during pregnancy, where it is found on fetal endothelial cells and on invasive cytotrophoblast cells, specifically those at the maternal / fetal interface. Its precise physiological role has yet to be defined. HLA-G may have nonimmune functions relating to angiogenesis and placentation, but most evidence suggests that it protects fetal cells from lysis by maternal uterine NK cells, which are found in large numbers around invading trophoblast cells. This effect is due to specific interaction with inhibitory receptors expressed on NK cells. We have examined the hypothesis that another function of HLA-G is to inhibit NK cell migration. Using an in vitro transmigration assay system, we present data to support this hypothesis. NK cell migration across porcine endothelial cells transfected with HLA-G1 was specifically inhibited compared to migration across HLA-A2-transfected monolayers. HLA- G1 had no influence on the migration of a control T lymphocyte line. These results support the idea that in vivo, HLA-G may inhibit NK cell traffic across the placenta. PMID- 10671216 TI - Heat shock proteins generate beta-chemokines which function as innate adjuvants enhancing adaptive immunity. AB - Heat shock proteins (HSP) are widely distributed and highly immunogenic molecules. A novel property reported here is that stimulation with HSP70 of CD8 enriched T cells derived from naive non-human primates caused a dose-dependent increase in concentrations of the beta-chemokines RANTES, macrophage inflammatory protein (MIP)-1alpha or MIP-1beta. However, the concentrations of these beta chemokines were greatly increased when the CD8 T cells derived from HSP70 immunized non-human primates were stimulated with HSP70. HSP linked to peptides or proteins combined generation of beta-chemokines with an adjuvant function by enhancing specific T cell proliferative responses and IgG and IgA antibodies. The beta-chemokine and adjuvant functions were also elicited by topical mucosal administration of HSP linked to an antigen. We postulate that microbial HSP can stimulate beta-chemokine production which may be responsible for innate adjuvanticity, as was found in cells eluted from normal rectal mucosal tissue, and constitutes a significant component of the mucosal-associated lymphoid system. Furthermore, stimulation of innate immunity may drive adaptive immunity and account for the protective effects of HSP against tumors and viruses. PMID- 10671217 TI - Xenopus NK cells identified by novel monoclonal antibodies. AB - Early-thymectomized (Tx) Xenopus frogs, which are permanently deficient in T cells, are used as a model sytem for the characterization of novel monoclonal antibodies (mAb) which identify candidate NK cells at the amphibian level of evolution. Hybridomas, generated from mice immunized with splenocytes from Tx Xenopus following B cell and thrombocyte depletion, were screened by flow cytometry. Three mAb (1F8, 4D4 and 1G5) were identified that stained increased proportions of splenocytes from Tx compared with control frogs. These mAb identified lymphoid populations from Xenopus spleen, liver and gut which, after 48 h culture in growth factor-rich medium, exhibited spontanous killing of MHC deficient allotumor targets. mAb-defined splenocytes also rapidly induced apoptosis of such tumor targets. Dual color analysis confirmed that NK cells are neither T nor B cells. Cytospins of splenocytes isolated with anti-NK mAb revealed large lymphoid cells with distinct pseudopodia. Immunohistology indicated each anti-NK mAb routinely labeled cells within the gut epithelium but NK cells were difficult to visualize in spleen sections. Western blotting of spleen, liver and intestinal lysates subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that 1G5 reacted strongly with protein bands of approximately 70 - 85 kDa, whereas mAb 1F8 and 4D4 stained less intensely, but identified similar protein bands. PMID- 10671218 TI - On the genetic mechanism of induction of CD3gamma-negative human T cell variants. AB - Invariant CD3gamma molecules are important components in TCR complex formation and function. Both CD3gamma alleles seem to be expressed co-dominantly. In the present report, we present experimental data which indicate that the induction of CD3gamma(-) Jurkat variants occurs with a frequency similar to that of TCRalpha( ) or TCRbeta(-) Jurkat cells. CD3delta(-), CD3epsilon(-) or CD3zeta(-) Jurkat variants were never obtained despite extensive efforts. Our data suggest a possible explanation for this genetic puzzle: the human CD3gamma gene has a mutational hot spot in a nucleotide sequence of nine adenosines (9A) in the exon 3 encoding most of the external CD3gamma domain. Thus, both CD3gamma alleles are easily mutated to either 8A or 10A sequences. Furthermore, absence of CD3gamma molecules in Jurkat T cells causes severe defects in TCR / CD3 assembly and function. PMID- 10671219 TI - Unaltered phenotype, tissue distribution and function of Valpha14(+) NKT cells in germ-free mice. AB - The expression pattern of mouse CD1d and the tissue distribution of CD1d restricted Valpha14-Jalpha281 NKT cells suggest that the liver and the marginal zone of the spleen might be preferred sites of activation of this potent innate pathway of early cytokine secretion. Because these tissues are particularly involved with the filtration of blood-borne pathogens, and because NKT cells with an activated / memory phenotype accumulate over the first weeks of life and their CD1 ligands bind microbial glycolipids, it has been hypothesized that expansion of the NKT cell subset may be driven by exposure to the microbial environment. To test this hypothesis, we analyzed the frequency, surface phenotype and functional properties of NKT cells in normal and in germ-free C57BL / 6 mice. Surprisingly, we found that the NKT cell subset develops in the presence or absence of a microbial environment. Although these results do not rule out the possibility that NKT cells exert a protective function against some microbial agents, they demonstrate that non microbial ligands, possibly self-antigens are sufficient for the generation, maturation and peripheral accumulation of NKT cells. PMID- 10671220 TI - Isolation of the intact white pulp. Quantitative and qualitative analysis of the cellular composition of the splenic compartments. AB - The spleen is anatomically and functionally divided into two compartments: the red pulp, where particles are effectively removed from the blood, and the white pulp, where specific immune responses are generated. Here the isolation of white pulp from red pulp is described, allowing a detailed analysis of the cellular components of both red and white pulp separately. A striking abundance of memory T cells was found in the white and red pulp with an overall ratio of T and B cells in the white pulp being similar to that in lymph nodes. Both NK and gamma delta T cells can be found in white pulp and lymph nodes, but granulocytes are absent. The distribution of dendritic cell subsets showed significant differences between white pulp and lymph nodes. Furthermore, short-term homing experiments showed that migration of lymphocytes into the white pulp greatly exceeded that into lymph nodes, with significant differences in migration of various lymphocytes subsets. This suggests a different migration and retention mechanism in the white pulp. This new isolation technique will allow further analysis of the functional capacities of the splenic compartments. PMID- 10671221 TI - Conversion of p56(lck) to p60(lck) in human peripheral blood T lymphocytes is dependent on co- stimulation through accessory receptors: involvement of phospholipase C, protein kinase C and MAP-kinases in vivo. AB - Activation of T cells requires co-stimulation of the TCR and accessory receptors like CD2, CD4, CD8, CD11a or CD28. Engagement of the TCR without co-stimulation results in anergy / apoptosis. Here we show that induction of the shift of the tyrosine kinase p56lck from 56 kDa to apparent 60 kDa in resting human peripheral blood T cells (PBT) is strictly dependent on co-stimulation through both TCR and accessory receptors. In contrast, triggering of the TCR alone is only sufficient to induce the lck shift in preactivated cells like T cell clones or the T lymphoma line Jurkat. Our studies predict an involvement of a phospholipase C isoform which surprisingly acts downstream of a phorbolester-sensitive, H7 insensitive protein kinase C. Inhibition of the lck shift in vivo by U73122, a specific inhibitor of phospholipase C, correlates with reduced activation of the MAP-kinases ERK1 / 2. Moreover, the MEK1-specific inhibitor PD98059 blocks the lck shift in vivo. These findings demonstrate that activation of the MEK1-ERK1 / 2 pathway is required for lck conversion in vivo. The lck shift is not inducible by co-stimulation through acidic sphingomyelinase or ceramides which even prevent ERK2 activation in PBT. Moreover, it is resistant to treatment with W7, KN62 and cyclosporin A. PMID- 10671222 TI - CD69-triggered ERK activation and functions are negatively regulated by CD94 / NKG2-A inhibitory receptor. AB - CD69 represents a functional triggering molecule on activated NK and T cells, capable of inducing cytotoxic activity and costimulating cytokine production. It belongs to the C-lectin type superfamily, and its gene maps in the NK gene complex, close to other genes coding for NK receptors. CD94 / NKG2-A complex is the inhibitory receptor for the non classical MHC class I molecule HLA-E on human NK cells. To investigate CD69-initiated signal transduction pathways, and to evaluate CD94 / NKG2-A interference on CD69 triggering ability, we have generated transfectants expressing both receptors in the RBL cell line. Here we report that CD69 engagement leads to the activation of extracellular signal-regulated kinase (ERK) enzymes belonging to the MAPK family, and that this event is required for CD69-mediated cell degranulation. Moreover, we show that the co-engagement of CD94 / NKG2-A inhibitory receptor effectively suppresses both CD69-triggered cell degranulation in RBL transfectants, through the inhibition of ERK activation, and CD69-induced cytotoxicity in human NK cells. Thus, here we provide new information on the molecular mechanisms initiated by CD69 activation receptor, and show that CD69-initiated signaling pathways and functional activity are negatively regulated by CD94 / NKG2-A inhibitory complex. PMID- 10671223 TI - A crucial role for p80 TNF-R2 in amplifying p60 TNF-R1 apoptosis signals in T lymphocytes. AB - Tumor necrosis factor-alpha (TNF-alpha) can elicit many cellular responses including programmed cell death or apotosis. TNF-alpha-induced apoptosis has been largely attributed to the p60 TNF-R1 receptor. The role of p80 in TNF-alpha mediated apoptosis is largely unknown. We now present evidence that signaling through p80 switches on the previously dormant apoptotic machinery associated with p60. This effect on p60-associated apoptosis involves the proximal activation of caspases and proceeds in the presence of strong NF-kappaB induction. We evaluated the role of TRAF2 in p80-assisted apoptosis and found that a decrease in TRAF2 protein occurs with p80 but not p60 stimulation. However, the decrease in TRAF2 protein can be dissociated from apoptosis in the presence of a caspase inhibitor. Hence, one means by which p80 TNF-R2 regulates apoptosis is through its ability to amplify internally apoptotic signal transduction from p60 TNF-R1. PMID- 10671224 TI - T cell activation-induced and HIV tat-enhanced CD95(APO-1/Fas) ligand transcription involves NF-kappaB. AB - CD95(APO-1 / Fas) ligand (CD95L) gene expression is critically involved in activation-induced T cell apoptosis. We and other have previously shown that HIV 1 Tat which is essential for efficient HIV gene expression sensitizes CD95 mediated apoptosis and up-regulates CD95L expression in T cells. In the present study we have investigated the regulatory mechanism for CD95L expression. Two NF kappaB binding sites are localized at - 537 to - 521 and - 57 to - 47 (relative to the transcription start site) of the human CD95L promoter. We show that both elements bind to NF-kappaB and SP-1 transcription factors and NF-kappaB is involved in transactivation of the CD95L promoter upon T cell activation. Mutations at each NF-kappaB site by two base pair substitutions resulted in 30 - 70 % reduction of the promoter activity. The effect of Tat on the human CD95L promoter activity was mapped to the same sites. Mutation of each NF-kappaB site also impaired the effect of Tat on CD95L promotor activity. We also show that ectopic expression of Tat protein in Jurkat T cells greatly increases NF-kappaB binding to its target DNA. Our studies provide evidence that Tat-enhanced CD95L expression is regulated at least in part by the NF-kappaB sites of the promoter. PMID- 10671225 TI - Effect of pre-existing cytotoxic T lymphocytes on therapeutic vaccines. AB - Therapeutic vaccines which aim to induce CD8(+) cytotoxic T lymphocyte (CTL) responses will often be required to perform in the presence of pre-existing CTL which recognize epitopes within the vaccine. Here we explore the ability of a viral vaccine vector presenting several co-dominant CTL epitopes to prime CTL responses in animals that have a pre-existing CTL response to one of the epitopes in the vaccine. The vaccine was usually capable of inducing multiple new responses, suggesting that immunodomination effects of pre-existing CTL may generally be minimal following vaccination. However, when large numbers of pre existing CTL were present, a novel type of immune modulation was observed whereby (1) the vaccine failed to prime efficiently new CTL responses that were restricted by the same MHC gene as the pre-existing responses, and (2) vaccine induced CTL responses restricted by other MHC genes were enhanced. These results may have implications for therapeutic multi-epitope vaccines for diseases like HIV and melanoma, which aim to broaden CTL responses. PMID- 10671226 TI - Immune down-regulation and peripheral deletion of CD8 T cells does not require TNF receptor-ligand interactions nor CD95 (Fas, APO-1). AB - TNF receptor-ligand interactions and CD95 (Fas / APO-1) have been demonstrated to be involved in activation-induced death of mature T cells. Here, we examined the role of these molecules in the murine model of lymphocytic choriomeningitis virus (LCMV) infection using LCMV TCR transgenic (tg) mice lacking TNF, TNF receptor I (TNFR1), CD95 or both TNFR1 and CD95. This report demonstrates that neither TNF receptor-ligand interactions nor CD95 was required for down-regulation of LCMV specific CD8 T cells following acute LCMV infection in vivo. Even LCMV-specific CD8 T cells lacking both TNFR1 and CD95 molecules declined after the acute phase of the infection with normal kinetics. Furthermore, peripheral deletion of LCMV specific CD8 T cells induced by LCMV peptide injection or by adoptive transfer of tg spleen cells expressing the corresponding LCMV epitope was not impaired in mice lacking TNF, TNFR1 and / or CD95. Our data speak against an indispensable role of these molecules in antigen-induced apoptosis of CD8 T cells in vivo and suggest that T cell homeostasis after antigen challenge is controlled by additional mechanisms. PMID- 10671227 TI - TNF receptor 1 (TNFR1) and CD95 are not required for T cell deletion after virus infection but contribute to peptide-induced deletion under limited conditions. AB - Deletion of mature T cells maintains cellular homeostasis and is involved in the maintenance of self tolerance to some peripheral self antigens. Previous studies have presented conflicting evidence for a role of the tumor necrosis factor receptor (TNFR) family member CD95 (Fas) in peripheral T cell deletion using CD95 deficient mice. To evaluate cooperation between CD95 and another TNFR family molecule, TNFR1, we generated mice deficient for both CD95 and TNFR1. We showed that TNFR1 and CD95 do not contribute to the decline of antigen-specific cytotoxic T lymphocytes after virus infection. Using TNFR1 / CD95-deficient mice expressing the P14 TCR specific for a lymphocytic choriomeningitis virus-derived peptide (p33) we showed that deletion of p33-specific CD8(+) T cells following high dose p33 administration is also normal. However, in non-TCR-transgenic TNFR1 / CD95-deficient mice treated with the same p33 regimen, tolerance induction was defective. These data indicate that TNFR1 and CD95 are dispensable for deletion of antigen-specific T cells after viral infection. However, under certain conditions, both TNFR1 and CD95 appear to cooperate in CD8(+) T cell deletion. PMID- 10671228 TI - B cell clonal elimination induced by membrane-bound self-antigen may require repeated antigen encounter or cell competition. AB - Transgenic mouse experiments indicate that autoreactive B cells are eliminated upon encounter with membrane self-antigen. In this study we tested how B cell tolerance to MHC class I antigens is affected by altering the frequency of antigen-carrying cells in mixed bone marrow (BM) chimeras. When antigen-bearing cells are present at low frequency, the reactive B cells and their antigens may coexist in the peripheral lymphoid organs, but under these conditions the B cells are functionally anergic and have a shortened lifespan. Such putative anergic cells are strongly deleted in the presence of additional, non-antigen-bearing, non-transgenic B cells. Since the antigen concentration on the surface of each antigen-bearing cell should be high, these results suggest that for efficient deletion of autoreactive B cells multiple antigen encounters may be required, particularly when cellular competition is weak. These results have implications for the therapeutic use of BM chimerism to induce B cell tolerance to grafts. PMID- 10671229 TI - Molecular characterization of two novel C-type lectin-like receptors, one of which is selectively expressed in human dendritic cells. AB - We have identified two human C-type lectin-like receptors, CLEC-1 and CLEC-2. Both display a single carbohydrate recognition domain and a cytoplasmic tyrosine based motif. They are homologous to the NK cell receptors NKG2s and CD94 and also to the oxidized low-density lipoprotein receptor 1. CLEC-1 and CLEC-2 are preferentially transcribed in dendritic cells (DC) and in the liver, respectively. Following transient transfection in COS cells, CLEC-1 is expressed intracellularly, perhaps requiring an associated chain to reach the cell surface. CLEC-2 is expressed on the surface of transfected cells as a protein of approximately 33 kDa. CLEC-1 and CLEC-2 genes map to human chromosome 12, most likely in linkage with the NK gene complex (NKC). Thus, the NKC may encode C-type lectin-like receptors expressed not only in NK cells but also in other cells, and at least one of these is of potential importance in regulating DC function. PMID- 10671230 TI - Dendritic cell maturation is induced by mycoplasma infection but not by necrotic cells. AB - To identify environmental stimuli that induce dendritic cell (DC) maturation, we exposed human monocyte-derived immature DC to apoptotic or necrotic cells and measured the levels of expression of costimulatory molecules and cytokine production. While most necrotic or apoptotic cells did not have any effect, some induced DC maturation as detected by up-regulation of CD83 and B7.2 and production of IL-12 and IL-6. The capacity of these cell lines to induce DC maturation was due to their contamination by mycoplasma, since the maturation inducing effect disappeared when the cells were treated with cyproxin. Furthermore, cell lines deliberately infected with mycoplasma containing supernatant acquired the capacity to induce DC maturation. Our results reveal that DC are able to sense mycoplasma infection and mature as they do in response to most viruses and bacteria. In contrast, apoptotic or necrotic cells fail to induce DC maturation. PMID- 10671231 TI - Activation of Cdk2 is a requirement for antigen-mediated thymic negative selection. AB - Apoptosis plays a critical role in T cell development and thymic selection. Thymocytes which undergo antigen-induced negative selection have been demonstrated to die by apoptosis. Despite this, relatively little is known about the specific apoptotic pathway involved in negative selection. We have examined the role of cyclin-dependent kinase 2 (Cdk2), a key regulator of thymocyte apoptosis, in this process. Stimulation of thymocytes with cognate antigen leads to a large increase in Cdk2 kinase activity. We also show that pharmacological inhibitors of Cdk2 block thymocyte apoptosis in response to antigen. Our data show that Cdk2 activity is essential for the apoptotic pathway used in negative selection. PMID- 10671232 TI - Impact of in utero Th2 immunity on T cell deviation and subsequent immediate-type hypersensitivity in the neonate. AB - It was the aim of this study to analyze the impact of maternal Th2 immune responses on onset and subsequent development of allergen-specific immunity and immediate-type hypersensitivity in early childhood. In a well characterized mouse model of Th2 immunity, BALB / c mice were sensitized to ovalbumin (OVA) before mating followed by allergen aerosol exposure during pregnancy. At the end of pregnancy mice developed allergen-specific Th2 / Th0 immunity and immediate-type hypersensitivity responses to OVA. T cells from these newborns, when restimulated with PMA / ionomycin, demonstrated a lowered capacity to produce IFN-gamma. To assess whether prenatal allergen exposure favors postnatal onset of a Th2-type immune response, these offspring were immunized to a novel antigen by a single injection of beta-lactoglobulin (BLG). In contrast to offspring from non sensitized mothers, offspring from OVA-sensitized mice showed both higher anti BLG immunoglobulin titers and higher frequencies of immediate-type skin test responses. Our data suggest that Th2 / Th0 immunity present during pregnancy has a decisive impact on shaping of the Th1 / Th2 T cell profile in the neonate. Furthermore, this effect favors the development of Th2 immune responses, when mice are exposed to a novel antigen during early childhood. PMID- 10671233 TI - Post-transcriptional regulation of E2A proteins via lipopolysaccharide and CD40 signaling. AB - The transcription factor E2A plays a crucial role in B cell development, the control of immunoglobulin gene rearrangement and expression. Here we report that in primary mouse B cells lipopolysaccharide (LPS) is able to induce the level of E2A protein by over 50-fold in days of culture. In contrast, CD40 signaling is insufficient to cause an E2A increase and can in fact prevent the LPS-mediated induction of E2A. These results suggest that E2A induction requires both proliferation and differentiation. We find that E2A protein induction is regulated post-transcriptionally since E2A mRNA is not induced by LPS. We have thus identified an important additional layer of regulation affecting the activity of E2A transcription factors. PMID- 10671235 TI - Highlights in the Renaissance of Amidometal Chemistry. AB - Turning a disadvantage to an advantage: The rather disappointing reactivity of the metal - nitrogen bond in comparison to the metal - carbon bond resulted in the neglect of amidometal chemistry after its heyday at the end of the 1960s and beginning of the 1970s. Today it is precisely this disadvantage which is being applied through the use of amido ligands to produce inert complex fragments with well-defined reaction centers. In this way the chemistry of the early transition metals has been markedly enriched, and interesting alternatives to the classically regarded cyclopentadienyl ligands are now available. PMID- 10671234 TI - Dissecting the Chemistry of Protein Splicing and Its Applications. AB - Protein splicing, the protein equivalent of RNA splicing, is a newly discovered posttranslational process that proceeds through a branched protein intermediate and produces two separate polypeptides from one gene. The experimental data used to distinguish among the proposed protein-splicing mechanisms are presented along with the progress made towards fully describing the mechanism. Numerous protein engineering applications using modified inteins have been developed, including the generation of alpha-thioesters in proteins, which circumvent the limits of solid-phase peptide synthesis. PMID- 10671237 TI - "Breaking the Rules": A Planar Phosphonium Cation. AB - Long-standing tenets of a discipline must be questioned in an effort to fully understand the fundamentals of science. This is exemplified by the synthesis of planar-tetracoordinate compounds, such as the square-planar phosphonium ion 1 by Driess et al., which provide exceptions to van't Hoff - Le Bel rules. PMID- 10671236 TI - How Long Have Nonlinear Effects Been Known in the Field of Catalysis? AB - The similarities and differences between nonlinear effects in asymmetric synthesis (see diagram), predicted 60 years ago by W. Langenbeck, and the long known amplification phenomenon in stoichiometric reactions with chiral starting materials are discussed. PMID- 10671238 TI - How Many Networks Can Be Made from Four-Coordinate Atoms? AB - In how many ways can three-dimensional space be divided into tiles such that four edges meet at every vertex? A procedure to calculate the number of possibilities was recently presented by the mathematician A. Dress. Some of the tilings correspond to the structures of zeolites and other networks. PMID- 10671239 TI - How to Make Drugs Orally Active: A Substrate Template for Peptide Transporter PepT1. AB - By building key structural features into hydrophilic drugs, they can be recognized by the PepT1 transporter system of the small intestine and rendered orally active. The model shown provides, for the first time, a 3D template for all known substrates of PepT1. PMID- 10671240 TI - Cholesteric Solid Films Formed by Spin-Coating Solutions of Dicholesteryl Esters. AB - Helical ordering of molecules is shown by solid films formed by spin-coating solutions of nonpolymeric, high molecular weight dicholesteryl esters (shown schematically). In these three-dimensionally organized films the axes of the helices are perpendicular to the surface of the glass plate. PMID- 10671241 TI - Al(30): A Giant Aluminum Polycation. AB - Simple hydrothermal treatment of the well-known aluminum polycation varepsilon Al(13) produces the novel Al(30) structure (see picture), the largest polycation yet observed. Its characterization, by X-ray diffraction and NMR spectroscopy, also solved previously unassigned signals in (27)Al NMR spectra of other Al - O species. PMID- 10671242 TI - The Vibrational Inelastic Neutron Scattering Spectrum of Dodecahedrane: Experiment and DFT Simulation. AB - Vibrational frequencies that are forbidden in Raman and IR absorption can be observed by inelastic neutron scattering. In the case of the I(h)-symmetrical molecule dodecahedrane (shown schematically), the resulting spectrum agrees with a DFT calculation. PMID- 10671243 TI - Supramolecular Materials with Electroactive Chemical Functions. AB - A diblock coil with diphenylaminophenyl side chains, oligoethylene oxide segments, and a rigid molecular pentamer of phenylene vinylene are present in the molecule studied here (see formula). When films of this material were investigated, the material was found to organize into nanostructures 12 nm in dimension. PMID- 10671244 TI - Kinetic Footprinting of an RNA-Folding Pathway Using Peroxynitrous Acid. AB - Following footprints to discover a path is easier with peroxynitrous acid. The folding of the Tetrahymena ribozyme was studied using this readily available reagent to generate hydroxyl radicals for kinetic footprinting studies. The different domains of the ribozyme appear to assemble at different rates following an ordered, hierarchical pathway (see scheme). PMID- 10671245 TI - Solvation of the Carbonyl Compound as a Predominant Factor in the Diastereofacial Selectivity of Nucleophilic Addition. AB - Temperature-dependent selectivity in nucleophilic additions is affected by the solvent. The inversion temperature (marked with arrows in the graph) that appears in the nonlinear Eyring plots of ln (anti/syn) versus temperature for the addition of butyllithium to an O-protected alpha-hydroxy aldehyde 1 does not depend on nucleophiles (nBuLi (black triangle), tBuLi (*)), but on the solvent. Its value can be obtained from a plot of the (13)C NMR chemical shift of C=O versus temperature. TBDMS=tBuMe(2)Si. PMID- 10671246 TI - Novel Single- and Double-Layer and Three-Dimensional Structures of Rare-Earth Metal Coordination Polymers: The Effect of Lanthanide Contraction and Acidity Control in Crystal Structure Formation. AB - Lanthanide atom sizes (the lanthanide contraction) directly control the type of structure formed by the coordination of a single multidentate ligand, 3,5 pyrazoledicarboxylic acid (H(3)pdc). Single-layer, double-layer ([Eu(2)(Hpdc)(3)(H(2)O)(6)], see picture), and three-dimensional networks were found. Control of the reaction pH plays a key role in the structure formation in this system. PMID- 10671247 TI - Synthesis of an alpha-(2,3)-Sialylated, Complex-Type Undecasaccharide. AB - Only two building blocks were necessary to provide efficient access to the complex undecasaccharide 1 by means of two stereo- and regioselective glycosylation reactions. Stereocontrolled p-methoxybenzyl-assisted beta mannosylation afforded the critical beta-mannoside linkage in 1, which is a constituent of the human glycoproteins follitropin and gonadotropin. PMID- 10671248 TI - Construction of Heterometallic Cubanes AB - Incorporation of M(CO)(3) fragments by trinuclear Ti complexes [{Ti(3)Cp(u(3)-CR)}(u-O)(3)] and [{Ti(3)Cp(u(3)-N)}(u-NH)(3)] (Cp*=eta(5) C(5)Me(5)) leads to the formation of an unprecedented class of heterometallic clusters with cubane structure [e.g., Eq. (a)]. Density functional calculations on these complexes indicate the existence of electron delocalization in the Ti(3)M cores (M=Cr, Mo, W). PMID- 10671249 TI - Coupling of Alkynes on a Pd-Pd Bond to Generate an Electrophilic u Butenediylidene Moiety. AB - The first dipalladium u-PPh(3) complex (1) was obtained by the facile loss of two CH(3)CN ligands from [Pd(2)(PPh(3))(2)(CH(3)CN)(4)](PF(6))(2) in CH(2)Cl(2). Coupling of p-tolylacetylene (p-TolC identical withCH) with 1 or its precusor afforded the u-butenediylidene complex 2, treatment of which with CH(3)CN resulted in PPh(3) migration to give 3. PMID- 10671250 TI - Evidence for an Equilibrium between an N-heterocyclic Carbene and Its Dimer in Solution. AB - Evidence for the Wanzlick equilibrium between carbene 1 and dibenzotetraazafulvalene (1)(2) at ambient temperature has been found (see scheme). Enetetramines of type (1)(2) can also be cleaved by coordinatively unsaturated transition metal compounds to form dicarbene complexes. PMID- 10671251 TI - Chromophore Alignment in a Chiral Host Provides a Sensitive Test for the Orientation - Intensity Rule of Induced Circular Dichroism. AB - Different inclusion modes can be realized for a bicyclic azoalkane in beta cyclodextrin (see picture). Variations in the alignment of the azo chromophore relative to the host can be detected through NMR spectroscopy and induced circular dichroism. The latter technique produces a positive signal for the lateral conformer and a negative, significantly weaker peak for the frontal one. This is in keeping with the orientation-intensity rule. PMID- 10671252 TI - Macrocyclic Imidazolylboranes. AB - Cycles, not chains: 1-imidazolylboranes exist in polymeric form as a consequence of donor/acceptor interactions. Through the use of suitable substituents and under high dilution tetrameric and pentameric macrocyclic imidazolylboranes were synthesized. The picture shows the structure of the tetrameric 4,5 dimethylimidazolylborane (black: C, gray: B, white: N). PMID- 10671253 TI - Nitrophenolate as a Building Block for Lanthanide Chains and Clusters. AB - Infinite, extended chains or tetradecanuclear lanthanide clusters (see structure) can be formed, depending on conditions, from the reaction of potassium o nitrophenolate with different lanthanide trichlorides. PMID- 10671254 TI - Molecular Recognition of Carbohydrates by Artificial Polypyridine and Polypyrimidine Receptors. AB - Despite their acyclic structure, the simple host molecules 1 and 2 can effectively hydrogen bond to monosaccharides. They show marked binding affinities to glucopyranosides in chloroform, and they are able to participate in three dimensional recognition of monosaccharides. PMID- 10671255 TI - Sequential Nucleophilic Substitution: A Powerful Strategy for the Solid-Phase Production of Diverse Compound Libraries. AB - Polyfunctionalized, unprotected reagents (e.g. amines and thiols) can be used in the production of highly diverse compound libraries by performing sequential nucleophilic substitutions on support-bound polyelectrophiles (see scheme). The procedure reported here enables the efficient preparation of new beta-alanine derivatives which are suitable for lead discovery. E*=polyelectrophile, *=solid support. PMID- 10671256 TI - New Solvatochromic Dyes of the 5-Dimethylamino-5'nitro-2,2'-bithiophene Type. AB - The first serious competitor to the known solvatochromic pyridiniumphenolate betaines has been uncovered with the 5-dimethylamino-5'-nitro-2,2'-bithiophene 1 a (X=Z=S, Y=CH, R=CH(3)), which has the added benefit of being notably sensitive to solvent in acidic media. By simple heterocyclization further compounds of the general type 1 have been synthesized, and their suitability as solvent indicators was investigated. In this way a new record was set with the selenophene analogue of 1 a. PMID- 10671257 TI - Doubly meso-beta-Linked Diporphyrins from Oxidation of 5,10,15-Triaryl Substituted Ni(II)- and Pd(II) - Porphyrins. AB - Lower oxidation potentials than for the monomeric starting materials are displayed by the diporphyrins obtained by one-electron oxidation with tris(4 bromophenyl)ammonium hexachloroantimonate [see, for example, Eq. (1)]. This and the strong red shift observed for the Soret bands of the product are indicative of extensively delocalized pi-electron systems in the fused diporphyrin. Ar=3,5 di-tert-butylphenyl. PMID- 10671258 TI - [Ni] AB - How do [NiFe] hydrogenases activate H(2)? Hydrogenases are key enzymes in the biological hydrogen and energy metabolism; the mechanism of H(2) activation, however, is unclarified. In particular, the oxidation states of the metals involved are discussed controversially. The title complex demonstrates that a distorted diamagnetic Ni(II) center and thiolate donors are sufficient (see picture) to catalyze the key reaction of hydrogenases, the H(2) heterolysis. PMID- 10671259 TI - A New Synthetic Route to Unsymmetrical 1,2-Bis(phosphanyl)ethanes and 1,2 Arsanyl(phosphanyl)ethanes with and without a Stereogenic Center. AB - A one-pot reaction affords unsymmetrical 1,2-bis(phosphanyl)ethanes 2 and 1,2 arsanyl(phosphanyl)ethanes 3 from the cyclic sulfate 1 in high yield. Similarly, chiral 1,2-bis(phosphanyl)ethanes and 1,3-bis(phosphanyl)propanes could be obtained in enantiomerically pure form. R, R'=alkyl, phenyl. PMID- 10671260 TI - A Novel Polymer-Supported Scandium Catalyst Which Shows High Activity in Water. AB - Lewis acids in water on a polymer support (1) have been used, for the first time, as a catalyst. The organic solvent-free catalytic activity, first tested with allylation reactions with tetraallyltin, extended to other Lewis acid catalyzed reactions with a range of carbonyl compounds to afford the corresponding alcohol; reactions of aromatic, aliphatic, heterocyclic aldehydes and even ketones proceeded smoothly. Catalyst recovery was quantitative after simple filtration. PMID- 10671262 TI - Diastereoselective Ce(OiPr)(3)-Catalyzed Pinacol Couplings of Aldehydes. AB - Aliphatic aldehydes as well as aromatic aldehydes can be converted by means of cerium-catalyzed pinacol coupling [Eq. (1)]. The coupling reaction products can be isolated with good to very good diastereoselectivities in favor of rac isomers (rac:meso>88:12). R=alkyl, aryl. PMID- 10671261 TI - A New Variant of the Claisen Rearrangement from Malonate-Derived Allylic Trimethylsilyl Ketene Acetals: Efficient, Highly Enantio- and Diastereoselective Syntheses of (+)-Methyl Dihydroepijasmonate and (+)-Methyl Epijasmonate. AB - Complete chirality transfer occurs in the smooth Claisen rearrangement of the trimethylsilyl (TMS) ketene acetals, which were prepared from allylic malonates (R)-1 (R=pentyl, 2-(Z)-pentenyl). These are in turn accessible by enantioselective reduction/esterification or by enzymatic kinetic resolution. The cis configuration in (+)-3 was achieved by highly syn-selective epoxidation of (+)-2, followed by suprafacial 1,2-H migration. PMID- 10671263 TI - The First Functionalized 6,12-Diazatetrakishomocubanes. AB - The photodimerization of asymmetric 4-aryl-1,4-dihydropyridines results, totally unexpectedly, in the new 6,12-diazatetrakishomocubanes 1. This is in contrast to the previously observed [2+2] photocycloaddition reactions of symmetrical 4-aryl 1,4-dihydropyridines PMID- 10671264 TI - Formation of a Novel Cage Compound with a Pentacyclo AB - Abnormally long C-C single bonds are found in the polycyclic caged diol with a pentacyclo[6.3.0.1(4,11).0(2,6).0(5,10)]dodecane skeleton formed by photolysis (see scheme). This skeleton resembles the structure of diamantane, but instead of having six cyclohexane rings in a chair conformation it contains only two cyclohexane rings in a distorted chair conformation and four cyclopentane rings, which makes it more highly strained than diamantane. PMID- 10671265 TI - How Stable Are Epoxides? A Novel Synthesis of Epothilone B. AB - Remarkable stability of the oxirane function is displayed over a number of synthetic operations in a novel synthesis of the antitumor compound epothilone B (see scheme). The cis-epoxide, generated very early by dihydroxylation of an (E) olefin, was resistant to more than ten synthetic steps under a wide variety of reaction conditions. TBS=tert-butyldimethylsilyl. PMID- 10671266 TI - Identification of Toxic 2,4-Decadienal in Oxidized, Low-Density Lipoprotein by Solid-Phase Microextraction. AB - The oxidation of low-density lipoprotein (LDL) in vitro was studied by a combination of solid-phase microextraction and GC/MS. 2-trans,4-cis-2,4 Decadienal, which is strongly toxic in vitro, was detected as the early oxidation product. This compound is degraded further to hexanal and (by cyclization of 4 hydroxy-2-nonenal) to 2-pentylfuran. PMID- 10671267 TI - Oxidation of Linoleic Acid in Low-Density Lipoprotein: An Important Event in Atherogenesis. AB - Contrary to earlier views the main oxidation products of low-density lipoprotein (LDL) are derived from linoleic acid and not arachidonic acid, as determined by GC/MS investigations of the in vitro oxidation of LDL samples. A similar product spectrum, in which epoxyhydroxyoctadecenoic acids such as 1 and 2 have been identified for the first time, is obtained from minimally oxidized (that is, aged) LDL. Since this is still recognized by the LDL receptor, it is concluded that toxic oxidation products are introduced in endothelial cells in vivo and cause damage there. PMID- 10671268 TI - Radical Intermediates in the Jacobsen - Katsuki Epoxidation. AB - Insight into the controversial mechanism of the Mn - salen-catalyzed epoxidation of olefins is provided in a theoretical study based on density functional theory. The calculations suggest that radical species A, but not manganaoxetanes B, are likely candidates for viable intermediates. PMID- 10671269 TI - Molecular Design of Liquid-Crystalline Block Molecules: Semifluorinated Pentaerythritol Tetrabenzoates Exhibiting Lamellar, Columnar, and Cubic Mesophases. AB - Fluid assemblies of star-shaped molecules like 1 form a range of thermotropic liquid crystalline phases, and represent a borderline case between anisometric mesogens, surfactants, and block copolymers. The low molecular mass molecules (<5.5 kDa) consist of a semipolar central core and a shell of semifluorinated chains. PMID- 10671270 TI - New Building Blocks for the Design of Oligonuclear Copper Complexes Based on Amino Carbohydrates. AB - Remarkably high magnetic coupling and O(2)-activation ability analogous to that of catechol oxidase are characteristics of the first structurally defined, low symmetry oligonuclear copper complexes of tridentate beta-oxoenamine ligands based on amino carbohydrates (the structure of a bis(acetylbutenonylaminoglucosidato)bis(u-acetato)tricopper(II) complex is shown). PMID- 10671271 TI - Simple Synthesis of a Chlorin - Fullerene Dyad with a Novel Ring-Closure Reaction. AB - To model the photosynthetic reaction center, the chlorin - fullerene dyad 1 was synthesized in a one-pot reaction from a linear tetrapyrrole and C(60). It is the first dyad of this type which contains a fullerene and a chlorin unit, the chromophore of naturally occurring photosynthetic systems. PMID- 10671272 TI - The First Crystal Structure of a Germanium(II) Amide with a Germanium - Lithium Bond and Its Behavior Towards Oxygen and Water. AB - The Ge-Li bond in 1 reacts readily with water and oxygen to give 2 (Li/H exchange) and 3 (insertion), respectively. In both cases the partial charge and the oxidation state of the germanium atom changes. All compounds were characterized by X-ray crystallography. PMID- 10671273 TI - Crystal Design of Barium Sulfate using Double-Hydrophilic Block Copolymers. AB - Peach, peanut, fiber, and flower (see picture) crystal morphologies are achieved from the precipitation of simple minerals in the presence of specifically adsorbing polymers. These crystal design effects are illustrated using BaSO(4) and double-hydrophilic block copolymers, the latter featuring carboxylate, sulfonate, phosphonate, and aspartic acid groups. PMID- 10671274 TI - Three-Strand Conducting Ladder Polymers: Two-Step Electropolymerization of Metallorotaxanes. AB - Sequential polymerization of supramolecular metallorotaxane complexes results in three-stranded conducting ladder polymers. The internal polymer is "sandwiched" between the other two polymers to give molecular wires that are effectively insulated when the outer polymers are in their nonconducting state (shown schematically). The intermolecular conductivity can be mediated by the Cu(1+)/Cu(2+) redox couple. PMID- 10671275 TI - Polar Thin Films Produced by Phosphonium Liquid Crystals: Two-Dimensional Self Assembled Ionic Layers with Spontaneous Polarization. AB - Nonlinear optical properties are shown by the smectic phase formed by simple dialkyl phosphonium salts. The molecules form into layers, featuring ion pairs separated by insulating glassy alkyl coating to produce a spontaneous polarization (P(s)) in the sheets (see picture). Applications of this material are currently leading to the development of novel electrical and electrooptical devices. PMID- 10671276 TI - Global Chirality in Rigid Decametallic Ruthenium Dendrimers. AB - Metallodendrimers with ten chiral Ru centers have been prepared in a stereospecific fashion (see picture; *=chiral Ru(diimine)(3) center). These molecules are conformationally rigid and exhibit well-defined global topologies: some diastereomers exhibit macroscopically chiral structures, others show a disklike topology. This difference in global or tertiary structure is exemplified by differences in their colloidal behavior, as observed in electric birefringence measurements. PMID- 10671277 TI - On the Location of Li(+) Cations in the Fast Li-Cation Conductor La(0.5)Li(0.5)TiO(3) Perovskite. AB - Lithium cations sit in the center of the oxygen "windows" defined by the vertex sharing TiO(6) octahedra of the title compound (see picture), as shown from neutron diffraction data. The octahedra are tilted to optimize the bond distances between the atoms. The unusual coordination and the partially unoccupied sublattice (occupancy factors 1/6 and (1/2) for Li and La, respectively) account for the high mobility of the Li(+) cations. PMID- 10671278 TI - New Synthetic Technology for the Rapid Construction of Novel Heterocycles-Part 1: The Reaction of Dess - Martin Periodinane with Anilides and Related Compounds. AB - The unusual behavior of hypervalent iodine reagents, Dess - Martin periodinane and IBX, with an array of anilides leads to the formation of complex heterocycles in only one synthetic operation (see scheme). Furthermore, the substrates for these transformations are available in one step from readily available commercial building blocks. The mechanism by which these periodinanes interact with anilides is also explored. This exciting new class of chemical reactions was discovered during the course of the total synthesis of the CP molecules and leads to compounds which are relevant to chemical biology investigations and pharmaceutical research. PMID- 10671279 TI - New Synthetic Technology for the Rapid Construction of Novel Heterocycles-Part 2. The Reaction of IBX with Anilides and Related Compounds. AB - The unusual behavior of hypervalent iodine reagents, Dess - Martin periodinane and IBX, with an array of anilides leads to the formation of complex heterocycles in only one synthetic operation (see scheme). Furthermore, the substrates for these transformations are available in one step from readily available commercial building blocks. The mechanism by which these periodinanes interact with anilides is also explored. This exciting new class of chemical reactions was discovered during the course of the total synthesis of the CP molecules and leads to compounds which are relevant to chemical biology investigations and pharmaceutical research. PMID- 10671280 TI - Cationic Phosphonolipids Containing Quaternary Phosphonium and Arsonium Groups for DNA Transfection with Good Efficiency and Low Cellular Toxicity** AB - Replacing the ammonium polar head in cationic lipids 1 (A=N) by a phosphonium or an arsonium group (A=P, As) improves their properties as synthetic vectors for DNA transfection. The increased volume of the cationic head is supposed to modify the interactions of the vector with the solvent and DNA. PMID- 10671281 TI - Phosphane Sulfide/Octacarbonyldicobalt-Catalyzed Pauson - Khand Reaction Under an Atmospheric Pressure of Carbon Monoxide. AB - Convenient access to cyclopentenones is provided by catalytic intra- and intermolecular Pauson - Khand reactions in the presence of octacarbonyldicobalt and tributylphosphane sulfide (see scheme). In contrast to other reactions of this type, they proceed under mild conditions (70 degrees C, 1 atm CO), and commercially available [Co(2)(CO)(8)] can be used without further purification. PMID- 10671282 TI - Synthesis and Properties of an Overcrowded Silabenzene Stable at Ambient Temperature. AB - Solid-state, room-temperature stability for silabenzene 1 is granted by the bulky silane rendering kinetic inertness. Silabenzene 1 is synthesized in good yield from the crowded chlorosilane 2 with tert-butyllithium. The delocalized, aromatic nature of 1 was confirmed by NMR spectroscopy and the (1)J(Si-C) value, as well as calculations on an analogous silabenzene. PMID- 10671283 TI - Catalytic Intermolecular Pauson - Khand Reactions in Supercritical Ethylene. AB - Ethylene is not only a substrate, but also a solvent: Catalytic intermolecular Pauson - Khand reactions of terminal alkynes were carried out in supercritical ethylene to provide 2-substituted cyclopentenones in moderate to high yields [Eq. (1)]. Under these conditions, even a low pressure of CO (5 atm) is sufficient for the reaction to take place. PMID- 10671284 TI - The First Red Azo Lake Pigment whose Structure is Characterized by Single Crystal Diffraction. AB - A model for red azo pigment Ca4B was characterized structurally using synchrotron radiation. This highly anisotropic ladder structure represents a new structural class in azo pigment chemistry. The picture shows that the calcium atoms coordinate in a complex manner to three azo ligands (one terdentate, one bidentate, and one monodentate) and two water molecules simultaneously. PMID- 10671285 TI - Detection of Toxoplasma gondii soluble antigen, SAG-1(p30), antibody and immune complex in the cerebrospinal fluid of HIV positive or negative individuals. AB - Active infection by T. gondii was evaluated by immunoassay for soluble SAG-1 (p30), the major surface antigen from T. gondii, specific antibodies and immune complexes in human cerebrospinal fluid (CSF) samples. A total of 263 samples of CSF were collected from hospitalized patients presenting neurological disorders and analyzed for antibodies to HIV. Patients were divided into two groups: HIV positive (n = 96) or HIV negative (n =167). The results of the assays showed that 45% of all samples were positive for soluble SAG-1. Toxoplasma Ag/Ab immune complexes were detected in 19% of the CSF samples and 62% were positive for T. gondii- specific IgG. A combination of these assays in the presence of clinical findings consistent with active Toxoplasma infection may predict the presence of toxoplasmic encephalitis. Moreover, detection of soluble SAG-1 in the CSF of these individuals appears consistent with active infection. PMID- 10671286 TI - Diagnosis of plague and identification of virulence markers in Yersinia pestis by multiplex-PCR. AB - We have developed a procedure for the rapid diagnosis of plague that also allows the identification of prominent virulence markers of Y. pestis strains. This procedure is based upon the use of a single polymerase chain reaction with multiple pairs of primers directed at genes present in the three virulence plasmids as well as in the chromosomal pathogenicity island of the bacterium. The technique allowed the discrimination of strains which lacked one or more of the known pathogenic loci, using as template total DNA obtained from bacterial cultures and from simulated blood cultures containing diluted concentration of bacteria. It also proved effective in confirming the disease in a blood culture from a plague suspected patient. As the results are obtained in a few hours this technique will be useful in the methodology of the Plague Control Program. PMID- 10671287 TI - Partial chemical characterization of antigenic preparations of chromoblastomycosis agents. AB - Antigenic preparations (saline, methylic, metabolic and exoantigens) of four agents of chromoblastomycosis, Fonsecaea pedrosoi, Phialophora verrucosa, Cladophialophora (Cladosporium) carrionii and Rhinocladiella aquaspersa were obtained. Partial chemical characterization of these antigenic preparations was obtained by determination of the levels of total lipids, protein, and carbohydrates, and identification of the main sterols and carbohydrates. Methylic antigens presented the highest lipid contents, whereas metabolic antigens showed the highest carbohydrate content. Total lipid, protein, and carbohydrate levels were in the range of 2.33 to 2.00 mg/ml, 0.04 to 0.02 mg/ml and 0.10 to 0.02 mg/ml, respectively, in the methylic antigens and in the range of 0. 53 to 0.18 mg/ml, 0.44 to 0.26 mg/ml, and 1.82 to 1.02 mg/ml, respectively, in saline antigens. Total lipid, protein, and carbohydrate contents were in the range of 0.55 to 0.20 mg/ml, 0.69 to 0.57 mg/ml and 10.73 to 5.93 mg/ml, respectively, in the metabolic antigens, and in the range of 0.55 to 0.15 mg/ml, 0.62 to 0.20 mg/ml and 3.55 to 0.42 mg/ml, respectively, in the exoantigens. Phospholipids were not detected in the preparations. Saline and metabolic antigens and exoantigens presented hexose and the methylic antigen revealed additional pentose units in their composition. The UV light absorption spectra of the sterols revealed squalene and an ergosterol fraction in the antigens. The characterization of these antigenic preparations may be useful for serological evaluation of patients of chromoblastomycosis. PMID- 10671288 TI - PCR with Paracoccidioides brasiliensis specific primers: potential use in ecological studies. AB - The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis genes. This fungus as well as several other fungi, were grown and their DNA obtained by mechanical disruption and a phenol chloroform isoamylalcohol-based purification method. The DNA served for a PCR reaction that employed specific primers from two P. brasiliensis genes that codify for antigenic proteins, namely, the 27 kDa and the 43 kDa. The lowest detection range for the 27 kDa gene was 3 pg. The amplification for both genes was positive only with DNA from P. brasiliensis; additionally, the mRNA for the 27 kDa gene was present only in P. brasiliensis, as indicated by the Northern analysis. The standardization of PCR technology permitted the amplification of P. brasiliensis DNA in artificially contaminated soils and in tissues of armadillos naturally infected with the fungus. These results indicate that PCR technology could play an important role in the search for P. brasiliensis' habitat and could also be used in other ecological studies. PMID- 10671289 TI - Jorge Lobo's disease: experimental inoculation in Swiss mice. AB - Sixty-four isogenic Swiss mice were intradermically inoculated in both hind foot pads. The inocula, consisting of fungal suspensions from biopsies obtained from Jorge Lobo's Disease patients, had the total number of fungi and the viability index determined using a Neubauer chamber and the fluorescein diacetate-ethidium bromide technique (FD-EB), respectively. The animals were sacrificed at times ranging from ten days to eighteen months after inoculation. The cellular infiltrate, mainly consisting of macrophages containing fungi, increased progressively up to end of the study; however, no macroscopic alterations were observed in the inoculated feet. After nine months, small numbers of Langhans' giant cells started to appear in the infiltrate. A considerable number of fungi was observed at the end of the experimental period, but only a few were viable when stained by the FD-EB technique. This fact suggests that there is a multiplication of fungal cells, which are destroyed by the macrophages but remain in the tissue for a long time due perhaps to the difficulties in their elimination. These findings led us to conclude that in spite of the maintenance of the infection in these animals, Swiss mice cannot be considered an ideal model to study Jorge Lobo's Disease. However, the authors call attention to the possibility of other mouse strains being more susceptible to Paracoccidioides loboi. PMID- 10671290 TI - Nosocomial infection in long-term care facilities. A survey in a Brazilian psychiatric hospital. AB - Nosocomial infection among male patients in a public psychiatric hospital was studied and the definitions for use in long-term care facilities were employed for diagnosis. The overall nosocomial infection rate was 6.7 per 1,000 day inpatients; 55.6% of these infections were identified in the respiratory tract, 50% of them being respiratory viral diseases; 38.9% of the nosocomial infections involved the eyes, ears, nose, throat and mouth, and 5.6% involved the skin and soft tissues. The epidemiological characteristics and the main clinical alterations of these inpatients were also identified. PMID- 10671291 TI - Gamma (60)Co DL(50/30) of Biomphalaria glabrata (SAY, 1818). AB - The variation of resistance to (60)Co gamma-rays of Biomphalaria glabrata was studied. A population of 480 mollusks was observed during 30 days - distributed in 8 groups of snails isolated and 8 groups of snails in colonies - after exposure (30 snails per group per dose) to increasing doses of gamma radiation. Doses of 10, 20, 40, 60, 80, 160, 320 and 640 Gy from a Gamma-cell (60)Co irradiator, were applied to the test groups and two groups control (non irradiated) of snails - isolated and colony - were kept apart. After have been exposed, the snails were drew back to the aquaria where they were maintained before. The survival was estimated on a daily score of the alive animals in each group-dose, starting after the irradiation exposure day. As a result, the survival self-fertilization forms (DL(50/30) = 218.2 Gy) was found greater than in cross-fecundation forms. These data point to a low radio-resistance on the cross-fertilization forms - the sexual reproductive form - which is most found in nature. The lower radio-resistance of the cross-fertilization forms suggests the presence of some sex-linked hormonal factor related to this phenomenon. PMID- 10671292 TI - Listeria monocytogenes in renal transplant recipients. AB - Five cases of Listeria monocytogenes bacteremia were observed from April to December 1985, among renal transplant recipients from the same hospital in Sao Paulo, Brazil. The patients were adults (mean age: 40.6 years), and the basic complaint was fever, with no report of meningeal syndrome. Laboratory tests revealed the presence of two serovars, (1/2)a and 4b, which were classified into three lysotypes. The four strains of serovar 4b showed the same antibiotype, with resistance to cefoxitin, clindamycin, oxacillin and penicillin. PMID- 10671293 TI - Flt3 ligand (FL) and its influence on immune reactivity. AB - Flt3 (fms-like tyrosine kinase 3) ligand (FL) is a potent hematopoietic cytokine that affects the growth and differentiation of progenitor and stem cells both in vivo and in vitro. Its capacity to augment strikingly the numbers of dendritic cells (rare antigen-presenting cells that induce and regulate immune responses) in mice and humans has stimulated considerable interest in its value as an investigational tool and therapeutic agent. In this review, we survey the hematopoietic properties and immunobiology of FL, and examine its therapeutic potential. PMID- 10671294 TI - Cloning, characterization and genomic organization of LCC-1 (scya16), a novel human CC chemokine expressed in liver. AB - By homology search of expressed sequence tags (EST) in GenBank a novel member of the CC chemokine family was identified. The full-length sequence of this liver specific CC chemokine (LCC-1) predicted a mature protein of 97 amino acids with 31-48% identity to other CC chemokines. There was a characteristic amino acid C term extension when aligned with other chemokines. Northern blot analysis from a panel of human tissues revealed that LCC-1 mRNA expression is restricted to adult and fetal liver. Different polyadenylation results in two mRNA species of 1.5 kb and 0.5 kb in size. LCC-1 is constitutively expressed in human HepG2 hepatoma cells and is induced by hypoxic exposure. The promoter region of the LCC-1 gene contains potential HIF-1 binding sites. The EST for LCC-1 has been previously mapped to the CC chemokine cluster on human chromosome 17q11.2. The organization of the LCC-1 gene (scya16) into three exons interrupted by two introns is identical to that found for other members of the CC chemokine family. PMID- 10671295 TI - Effects of stimulus and cell type on the expression of the -308 tumour necrosis factor promoter polymorphism. AB - The authors have previously demonstrated that the tumour necrosis factor (TNF) 308 G/A polymorphism affects the binding of transcription factors. In transient transfection assays in PMA stimulated U937 monocytes and Jurkat T cells, the A containing TNF2 promoter has a 2-3-fold greater transcriptional activity than the TNF1 promoter in the presence of the TNF 3'UTR. In this study it was found that a difference in TNF1 and TNF2 promoter activities was only observed in U937 and Jurkat cells, and not in Raji (B cell line), HeLa (epithelial carcinoma cell line), HepG2 (hepatoma cell line) or THP-1 (monocyte), suggesting cell-type specific transcription factors or modifications may be involved in the formation of the -308 protein/DNA complex. Physiological stimulators, TNF and interferon gamma (IFN-gamma) did not cause differential promoter activity between TNF1 and TNF2, but LPS did with only the TNF2 promoter/3'UTR construct being significantly responsive to lipopolysaccharide (LPS) in U937 cells. In U937 cells, the -308 polymorphism affected transcription following differentiation by phorbol myristate acetate (PMA), retinoic acid, PMA plus LPS and PMA plus retinoic acid with an increase in nuclear factor binding to both TNF1 and TNF2 in the -323 to 285 region being observed. The greatest difference between TNF2 and TNF1 promoter activities (5-fold) was observed following PMA plus retinoic acid treatment of transfected U937 cells for 48h. During this time, U937 differentiated into cells with a macrophage-like morphology. An understanding of the cell type and stimuli specific requirements for differential expression of the -308 polymorphism may help elucidate the role the TNF -308 polymorphism plays in diseases where elevated TNF levels are thought to be important. PMID- 10671296 TI - Pseudoexons and regulatory elements in the genomic sequence of the beta chemokine, alternative macrophage activation-associated CC-chemokine (AMAC)-1. AB - Recently, the authors reported the cloning of a novel human CC chemokine of alternatively activated macrophages (AMAC-1), whose expression is induced by Th2 associated cytokines such as interleukin 4 (IL-4), IL-13 and IL-10; vice versa, AMAC-1 expression is inhibited by Th1-associated cytokines such as interferon gamma (IFN-gamma). In order to study the genomic organization and transcriptional regulation of the AMAC-1 gene, genomic clones were isolated by screening a human lambda genomic library. Sequencing of a clone with a 1.7-kb insert gave a partial genomic sequence for the AMAC-1 gene. The complete AMAC-1 genomic sequence was obtained by bioinformational methods and the whole region spanning the AMAC-1 gene was verified by PCR amplification of subfragments and sequencing. The AMAC-1 gene consists of three exons. Whereas exons 2 and 3 were separated by a small intron of 411 bp, exon 1 and exon 2 were separated by 6 kb of non-translated genomic sequence containing two pseudoexons that are not expressed although they feature intact exon/intron boundaries and complete open reading frames. In order to allow a detailed analysis, a 2.7-kb fragment containing the promoter region and the first exon of AMAC-1 gene was cloned into a reporter gene construct. In the AMAC-1 promoter, two possible transcription start points were identified. In addition, several putative regulatory sequences for IL-4- and IFN-gamma-dependent transcriptional pathways were found including STAT6 and STAT1 binding sites as well as several AP-1 and C/EBP elements. Interestingly, a combined STAT6/STAT1 binding element is located in the direct vicinity of the first putative transcription start point. Competitive binding of IL-4-induced STAT6 versus IFN gamma-induced STAT1 to this site may explain the antagonistic effects these cytokines exert on AMAC-1 expression. PMID- 10671298 TI - Regulated expression of alternate transcripts from the mouse oncostatin M gene: implications for interleukin-6 family cytokines. AB - Oncostatin M (OSM) is a member of the IL-6 family of polyfunctional cytokines. The characterized murine OSM transcript consists of three exons and encodes a secreted protein. Investigations of mOSM expression using the ribonuclease protection assay demonstrated novel sites of expression in undifferentiated but not differentiated pluripotent cells, and revealed the existence of alternatively spliced mOSM transcripts. cDNAs representing a novel mOSM transcript (mOSM 13) containing exon 1 spliced directly to exon 3 were isolated from bone marrow using Rapid Amplification of cDNA Ends (RACE) PCR and RT-PCR approaches. Expression of the mOSM 13 transcript was regulated in a tissue-specific manner and independently of mOSM transcript production, suggesting that its production is biologically significant. Splicing of exon 1 directly to exon 3 disrupts the OSM open reading frame of mOSM 13. Initiation of translation at sites within exon 3 of mOSM 13 would yield N-terminally truncated OSM proteins that are localized within the cell. The omission of exon 2 by alternate splicing and the production of intracellular proteins with alternate biological activities are conserved among several IL-6 family cytokines and are one manifestation of a more general phenomenon; the production of alternate cytokine transcripts encoding intracellular and extracellular proteins. PMID- 10671297 TI - Tyrosine 462 of the membrane-proximal F'-G' loop of murine Mpl is not essential for high-affinity binding of thrombopoietin. AB - The ligand binding site of Mpl, the thrombopoietin (Tpo) receptor, has not been determined. Tyr(462)of murine Mpl corresponds to Tyr(421)of the common beta chain of the human IL-3, IL-5 and GM-CSF receptors. Tyr(421)has been identified as essential for high-affinity ligand binding. To determine whether Tyr(462)is similarly required for Tpo binding, wild-type murine Mpl (Mpl-WT) or mutant receptors containing an alanine (Y462A) or lysine (Y462K) in place of Tyr(462)were expressed in BaF3 cells. In proliferation studies, the Y462A mutation had no effect on Tpo-induced growth. In contrast, the Y462K mutation led to an attenuated proliferative response to Tpo. In single-point binding studies, both Mpl-WT and Y462A cells were able to bind [(125)I]Tpo in a specific manner. In contrast, there was a marked reduction in binding of [(125)I]Tpo by Y462K cells. Mpl-WT cells bound Tpo with a K(d)of approximately 330 pM, while Y462A cells bound Tpo with a K(d)of approximately 268 pM. The binding affinity of Y462K cells was below that quantifiable by Scatchard analysis. This study suggests that unlike the corresponding Tyr(421)of the common human beta chain, Tyr(462)of murine Mpl is not required for high-affinity ligand binding, although it may be located in proximity to the ligand binding site. PMID- 10671299 TI - Tumour necrosis factor alpha enhances the expression of hydroxyl lyase, cytoplasmic antiproteinase-2 and a dual specificity kinase TTK in human chondrocyte-like cells. AB - Tumour necrosis factor alpha (TNF-alpha) is a cytokine with pleiotropic effects on cells ranging from proliferation to apoptosis. These biological effects of TNF alpha are believed to be elicited by the induction or enhancement of the expression of TNF-alpha responsive genes in the target cells. TNF-alpha is pro inflammatory and a principal mediator in the pathogenesis of arthritis. The activation of an inflammatory cascade by TNF-alpha in arthritis results in the degradation of cartilage, joint destruction and loss of function. Because TNF alpha is an important mediator in the pathogenesis of arthritis, the present study addresses the identification of novel TNF-alpha responsive genes in HTB-94 cell line which is of human origin and maintains a chondrocytic phenotype. The three identified cDNAs were previously not known to be induced or upregulated by TNF-alpha in chondrocytes or cells of chondrocytic lineage. One of the identified cDNAs had sequence similarity to human hydroxyl lyase mRNA (PLOD), an enzyme involved in collagen biosynthesis and its metabolism; the second cDNA had sequence similarity to the human cytoplasmic anti-proteinase-2 mRNA (CAP-2), a member of a group of proteins shown to be associated with protecting cells from TNF-alpha-induced apoptosis; and the third cDNA had sequence similarity to a dual specificity kinase, TTK, which is associated with cell proliferation. Relative gene expression level analysis by PCR and by Northern blotting revealed that treatment with TNF-alpha enhanced the expression of PLOD, CAP2 and TTK transcripts which confirmed the results obtained with display gels. Furthermore, TTK mRNA expression was also induced in human articular chondrocytes treated with TNF-alpha but not in untreated chondrocytes. Our results suggest that these genes may play a role in chondrocytic responses to TNF-alpha-mediated stimuli affecting the cartilage homeostasis. PMID- 10671300 TI - Soluble glycoprotein 130 (gp130) attenuates OSM- and LIF-induced cartilage proteoglycan catabolism. AB - Oncostatin M (OSM) and leukaemia inhibitory factor (LIF) exhibit pleiotropic biological activities and share many structural and genetic features. The two cytokines bind with high affinity to the same receptor (LIF/OSM receptor), which consists of the LIF receptor alpha chain (LIFRalpha) and the signal transduction unit gp130. A soluble form of the beta chain of the receptor complex called soluble gp130 (sgp130) has been cloned. In this study, we sought to determine whether recombinant sgp130 or anti-gp130 Ab could attenuate the resorption of proteoglycans induced by OSM and LIF in articular cartilage explants. The results show that at high concentrations sgp130 is capable of attenuating both LIF and OSM mediated resorption. In contrast, anti-gp130 Ab selectively inhibited the stimulation of proteoglycan (PG) release by OSM, albeit minimally. The failure of anti-gp130 to attenuate LIF stimulated PG resorption may be due to the normal interaction of LIF with LIFRalpha and unfettered heterodimerization of LIFRalpha with gp130 in the presence of the antibody. The results indicate that sgp130 and anti-gp130 can modulate cartilage PG metabolism in vitro. Whether sgp130 may have therapeutic activity in models of arthritis or indeed in arthritic diseases remains to be determined. PMID- 10671301 TI - Total serum IL-12 and IL-12p40, but not IL-12p70, are increased in the serum of older subjects; relationship to CD3(+)and NK subsets. AB - Interleukin 12 (IL-12), a central cytokine acting on T and natural killer (NK) cells, directs proliferation of activated T lymphocytes towards a Th1 phenotype. The heterodimeric molecule IL-12p70, equates with IL-12 biological activity, while IL-12p40 may antagonize IL-12 and inhibit cytotoxic T lymphocyte (CTL) generation in vitro. This study characterizes age-related changes in serum total IL-12, IL-12p70 and IL-12p40 relating them with CD3(+), NK and related subsets from subjects, aged 30-96 years. Total IL-12, IL-12p40 and the IL-12p40/IL-12p70 ratio, but not IL-12p70, increased significantly with age (P<0.0001). Increases in total IL-12 and IL-12p40 were negatively associated with CD3(+)(P=0.003, P=0.002), CD3(+)CD4(+)(P=0.004, P=0.003), CD3(+)CD8(+)(P=0.04;P=0. 04) and CD4(+)45RA(+)(P=0.0003;P=0.0007) subsets, respectively. Conversely, increases in IL-12p40 showed a non-significant trend for association with increases in NK(P=0.07) and a related CD8(+low)CD57(+)(P=0.07) subset. These findings may have important implications for understanding the functional activity of IL-12 and its p40 and p70 subunits in vivo and with respect to T-or NK-cell activation in aging. PMID- 10671302 TI - Analysis of soluble interleukin 6 receptor in cerebrospinal fluid in inflammatory and non-inflammatory conditions. AB - The objective of this study was to investigate the pathophysiological roles of soluble interleukin 6 receptor (sIL-6R) in cerebrospinal fluid (CSF). CSF was obtained from patients suspected with meningitis. Eight patients without any meningeal signs or symptoms were enrolled as controls. An additional 34 CSF samples were collected to measure both biologically active and immunoreactive sIL 6R. All CSF samples were proven to be aseptic. IL-6 and sIL-6R were measured using specific ELISAs. Patients were divided into three groups on the basis of cell number in CSF; inflammatory group (cell number >5 microl, mean 241+/-363.1, n=61); non-inflammatory group (cell number < or =5 microl, mean=2.1+/-1.7, n=12) and controls (cell number < or =5 microl, mean=0.3+1.7, n=8). Among these three groups, the differences in protein (F (2,78)=8.274, P<0.0001) and IL-6 concentration (F (2,78)=6.475, P<0.001) were statistically significant but those of sIL-6R concentration were not. There were only weak correlations between log (sIL-6R) versus log (cell number) (r=0.23, P=0.0375), log (protein) (r=0.239, P=0.0358) and log (IL-6) (r=0.27, P=0.0167). Amounts of immunoreactive and biologically active sIL-6R were closely correlated (r=0.62, n=34, P<0.005). It was concluded that sIL-6R is present constitutively in CSF and its level may not increase significantly in inflammatory conditions; infiltrating cells in CSF are not the main source of sIL-6R; and sIL-6R in CSF can bind IL-6. PMID- 10671303 TI - Randomized controlled trial of recombinant human granulocyte-macrophage colony stimulating factor for the treatment of chronic hepatitis c. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) was administered subcutaneously to 45 chronic hepatitis C patients, randomly assigned to receive 0.5, 1 or 2 microg GM-CSF/kg b.w. daily/6 weeks (n=30), or no treatment (n=15). Alanine transaminase (ALT) values normalized in four out of 10 (40%) patients administered 2 microg GM-CSF [1 cleared hepatitis C virus (HCV) RNA] but in none given 0.5 or 1 microg or untreated controls (P=0.0079). Following 4 weeks of rest, patients received 5 million units of interferon (IFN)alpha2b every other day/6 months, alone (n=30), or combined with 2 microg GM-CSF/daily for 3 months (n=15). At treatment end, ALT levels in patients administered the combination normalized more frequently than in those given monotherapy (73% vs 47%, P=0.089). Viraemia decreased significantly in 11/15 (73%) patients administered GM CSF/IFNalpha2b combination (mean log HCV RNA copies/ml+/-SEM: 4.13+/-0.40 vs 5.29+/-0.23;P=0.011), and in 20/30 (67%) receiving IFNalpha2b monotherapy (4.27+/ 0.28 vs 5. 31+/-0.14;P=0.004); 27% and 20% of patients given the combination and monotherapy, respectively, cleared HCV RNA. One patient in each regime had a sustained response after 12 months. 2', 5'-Oligoadenylate synthetase activity (2 5AS) increased during GM-CSF therapy (P=0.033 with the 2 microg dose). 2-5AS increased more in the GM-CSF/IFN-alpha2b combination than with IFNalpha2b monotherapy (P<0.02). GM-CSF provoked a skin reaction at the injection site, accompanied by moderate and reversible rises in eosinophil and leucocyte counts. In summary, daily s.c. GM-CSF administration is safe and shows effects against HCV; the GM-CSF/IFNalpha2b combination has an additional-but transient-antiviral activity in chronic hepatitis C. PMID- 10671304 TI - Linkage disequilibrium testing of four interleukin-1 gene-cluster polymorphisms in Danish multiplex families with insulin-dependent diabetes mellitus. AB - The molecules of the interleukin 1 (IL-1) system have been suggested to play a role in the pathogenesis of insulin-dependent diabetes mellitus (IDDM), and polymorphisms in the genes encoding IL-1beta (IL1B), the IL-1 Type 1 receptor (IL1RTI) and the IL-1 receptor antagonist (IL1RN) molecules have been associated with IDDM in case-control studies. It can be difficult to exclude selection biases in case-control studies leading to spurious association. This risk is eliminated when using the transmission disequilibrium test (TDT). Hence, by means of the TDT we have investigated four intragenic IL-1 gene-cluster polymorphisms, the IL1B AvaI, the IL1B TaqI, the IL1RTI PstI and the IL1RN 2(nd)intron polymorphisms, for linkage and intra-familial association with IDDM in Danish IDDM multiplex family material comprising 245 families. We found no evidence for overall linkage or intra-familial association between any of the polymorphisms and IDDM. In addition, we did not find linkage between any of the polymorphisms and IDDM in HLA-DR3/4 heterozygous or HLA-non-DR3/4 heterozygous IDDM subjects, respectively. In conclusion, by means of intra-familial TDT analysis we found no linkage or intra-familial association between IDDM and the four IL-1 gene-cluster polymorphisms in Danish IDDM multiplex family material. PMID- 10671305 TI - Placental cytokine expression in preterm labour and the fetal inflammatory response. PMID- 10671306 TI - Gap junctions. PMID- 10671307 TI - Use of antibodies in the analysis of connexin 43 turnover and phosphorylation. AB - A series of antipeptide antibodies designed to recognize specific sequences of the gap junction protein connexin 43 (Cx43) were developed and characterized immunochemically and immunohistologically. These antibodies bound to gap junctions and, on Western blots, to 43-kDa (often resolved as a doublet) and 41 kDa proteins in samples from heart, leptomeningeal cells, and brain. Relatively little of the 41-kDa protein was detectable in heart homogenates. Cultured rat leptomeningeal cells expressed high levels of the gap junction protein Cx43 and were used to analyze its turnover and phosphorylation. Pulse-chase experiments in leptomeningeal cells with [(35)S]methionine indicated that the 41-kDa form of connexin 43 was the first immunoprecipitable translation product. Radiolabel subsequently appeared in the lower band of the doublet at 43 kDa, followed by a shift into the higher band and turnover of the protein with a t(1/2) of 2.7 h. Pulse-chase labeling with [(32)P]P(i) indicated that phosphorylation of connexin 43 was limited to the 43-kDa protein, with a t(1/2) of 1.7 h. Treatment with alkaline phosphatase shifted the apparent molecular mass of the 43-kDa protein doublet such that it comigrated with the 41-kDa form. Hence, the 43-kDa protein observed on Western blots of both leptomeningeal cells and heart arises by phosphorylation of the 41 kDa precursor. Phosphorylation of serine residues accounts for most, if not all, of Cx43 phosphorylation in this system. PMID- 10671308 TI - Electron crystallographic methods for investigating gap junction structure. AB - Gap junctions are clusters of closely packed intercellular membrane channels embedded in the plasma membranes of two adjoining cells. The central pore of the membrane channels serves as a conduit between cell cytoplasms for molecules less than 1000 Da in size. Advances in the purification of gap junctions and electron cryocrystallography and computer reconstruction techniques have produced new insights into the intercellular channel structure. Methods are described here for the purification of gap junction membranes, biochemical treatments to produce hemichannel layers ("split junctions"), assessment of the purity of gap junction preparations, electron cryomicroscopy, image processing and reconstruction, three dimensional visualization, and interpretation. The critical step in electron crystallographic structure determination remains the isolation of crystalline material in sufficient and pure quantities for recording of electron microscope images. Along with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the quality of gap junction purification is assessed using electron microscopy of negatively stained preparations. Electron microscopy is also used to assess the crystallinity of the purified gap junctions and split junctions. Electron cryocrystallography is a powerful technique for high resolution structural characterization. Image processing is used to combine and enhance two-dimensional images. Electron crystallographic analysis is used to generate a three-dimensional structure from a set of electron micrographs. This three-dimensional information is extracted from a set of images recorded after tilting the specimen in the electron microscope stage and recombined using Fourier analysis techniques analogous to those used in X-ray crystallography. Computer modeling of the three-dimensional gap junction structures is a useful tool for analyzing hemichannel docking. PMID- 10671309 TI - Analysis of connexin intracellular transport and assembly. AB - Central to understanding the establishment and regulation of gap junction mediated intercellular communication is a detailed knowledge of how these complex structures are assembled. In this article, we describe methods to modulate and/or monitor the transport of connexins from the endoplasmic reticulum to the Golgi complex and from the trans Golgi network to the cell surface. We also present techniques that we have developed to assay assembly of newly synthesized connexin monomers into connexons and gap junctional plaques. PMID- 10671310 TI - Cell-free synthesis for analyzing the membrane integration, oligomerization, and assembly characteristics of gap junction connexins. AB - For gap junction channels to function, their subunit proteins, referred to as connexins, have to be synthesized and inserted into the cell membrane in their native configuration. Like other transmembrane proteins, connexins are synthesized and inserted cotranslationally into the endoplasmic reticulum membrane. Membrane insertion is followed by their assembly and transport to the plasma membrane. Finally, the end-to-end pairing of two half-channels, referred to as connexons, each provided by one of two neighboring cells, and clustering of the channels into larger plaques complete the gap junction channel formation. Gap junction channel formation is further complicated by the potential assembly of homo- as well as heterooligomeric connexons, and the pairing of identical or different connexons into homo- and heterotypic gap junction channels. In this article, I describe the cell-free synthesis approach that we have used to study the biosynthesis of connexins and gap junction channels. Special emphasis is placed on the synthesis of full-length, membrane-integrated connexins, assembly into gap junction connexons, homo- as well as heterooligomerization, and characterization of connexin-specific assembly signals. PMID- 10671311 TI - Connexin and gap junction degradation. AB - Many of the subunit proteins (connexins) that form gap junctions are rather dynamic, with half-lives of only a few hours. Thus, alterations in connexin turnover and degradation may represent significant mechanisms for the regulation of intercellular communication. We describe a pharmacological approach to determining pathways of connexin degradation. Cell cultures are left untreated or treated with inhibitors of lysosomal or proteasomal proteolysis. Changes in connexin levels, localization, or decay curves (derived from pulse-chase experiments) are assessed by immunoblotting, immunofluorescence, and immunoprecipitation, respectively. Such experiments have provided evidence that connexin43 degradation involves both the lysosome and the proteasome. PMID- 10671312 TI - Chemical gating of gap junction channels. AB - Chemical gating of gap junction channels is a complex phenomenon that may involve intra- and intermolecular interactions among connexin domains and a cytosolic molecule (calmodulin?) that may function as channel plug. This article focuses on the methodology we have employed for studying the molecular basis of chemical gating by lowered cytosolic pH. Our approach has combined molecular genetics and biophysics, using exposure to 100% CO(2) for assaying chemical gating efficiency. Chimeras of connexin 32 (Cx32) and connexin 38 (Cx38) and Cx32 mutants modified at residues of the cytoplasmic loop, the initial C-terminus domain, or both have been expressed in Xenopus oocytes, and channel expression and gating have been tested electrophysiologically by double voltage clamp. In addition, various channel forms, including homotypic, heterotypic, and heteromeric channel combinations, have been evaluated for chemical gating sensitivity. PMID- 10671313 TI - Analysis of connexin phosphorylation sites. AB - Most connexins, the proteins that form gap junction channels, are phosphoproteins. Connexin phosphorylation has been thought to regulate gap junctional protein trafficking, gap junction assembly, channel gating, and turnover. Connexin phosphorylation has been investigated in a variety of ways. Some connexins show mobility shifts in sodium dodecyl sulfate-polyacrylamide gel electrophoresis on phosphorylation. Kinase modulators can change the level of connexin phosphorylation and affect gap junctional communication levels. Metabolic labeling of cultured cells has allowed both phosphoamino acid identification and generation of phosphotryptic peptide maps. However, identification of the location of phosphorylated residues within the connexin sequence has required either targeted peptide synthesis, in vitro phosphorylation of known sites, and two-dimensional comigration studies or liquid chromatographic separation and N-terminal sequencing of peptides. In addition to these conventional methods, we discuss new applications of mass spectrometry to the identification of phosphorylated peptides and the specific residues phosphorylated within the connexin-derived peptide. PMID- 10671314 TI - General or cell type-specific deletion and replacement of connexin-coding DNA in the mouse. AB - Here we describe several gene targeting approaches currently used in our laboratory for the generation of deletion or replacement mutants of connexin genes in the mouse and discuss the advantage of the double-replacement strategy for the generation of conditional mutants. For the analysis of complementary functions of connexins, it will be necessary to generate mice with mutations in several connexin genes. We also report how this can be effectively accomplished. The replacement of targeted connexin-coding DNA with a reporter gene, to mimic expression of the deleted gene product, is currently being used in several laboratories. The use of different reporter genes or their differently localized gene products could allow distinction of promoter activity in double or triple connexin mutant mice. PMID- 10671315 TI - Identification of connexin-interacting proteins: application of the yeast two hybrid screen. AB - Protein-protein interactions are recognized as one of the fundamental mechanisms for relaying the intra- and intercellular signals that are required for normal cellular activities affecting growth, development, and maintenance of homeostasis in tissues and organs. The yeast two-hybrid screen has become a valuable tool for identifying protein-protein interactions. The gap junction protein connexin 43 (Cx43) has been implicated in a number of biological processes including development and cellular growth control. To further advance our understanding of the ways in which Cx43 may influence these cellular activities, and to extend our knowledge of the regulation of Cx43 function and/or processing, we have employed the yeast two-hybrid screen technique to identify Cx43-interacting proteins. We present detailed methods for the yeast two-hybrid screen of a mouse embryonic cDNA library using the C terminus of Cx43 as "bait." We also describe additional methods to confirm the interactions between Cx43 and the identified proteins. These methods include in vitro binding assays, coimmunoprecipitation, and subcellular localization using immunofluorescence microscopy. PMID- 10671316 TI - Probing the function of connexin channels in primary tissues. AB - Connexin channels provide for a widespread mechanism of cell-to-cell cross-talk within primary tissues, which is mediated by intercellular exchanges of cytoplasmic ions and molecules. Experimental and clinical studies have recently provided evidence that these exchanges are most likely to play multiple roles, which are critical for the proper development and function of primary tissues. There is also increasing evidence that major clinical disorders may result when the formation and function of connexin channels are altered. Still, the physiological functions that the cell-to-cell communication mediated by connexin channels subserve in most primary tissues are still uncertain. Here, I review two approaches that may aid in identifying these specific functions. PMID- 10671317 TI - Gap junctions and the regulation of cellular functions of stem cells during development and differentiation. AB - In multicellular organisms, the role of gap junction intercellular communication (GJIC) in the regulation of cell proliferation, cell differentiation, and apoptosis is becoming increasingly recognized as one of the major cellular functions from the start of the fertilized egg, through normal development of the embryo and fetus, to the sexual maturation of the adult and ultimately to the maintenance of health of the aging adult. Given that the function of this membrane-associated protein channel is to synchronize electrotonic or metabolic functions, differential regulation of function at the transcriptional, translational, and posttranslational levels of a family of highly evolutionarily conserved genes (connexins) needs to be considered. Both inherited mutations and environmental modulation of GJIC could, in principle, affect the function of gap junctions to control cell proliferation, cell differentiation, and apoptosis, thereby leading to a wide variety of pathologies. We review a few techniques used to characterize the ability of stem and progenitor cells to perform GJIC. PMID- 10671318 TI - Immunohistochemical analysis of cerebral cortical and vascular lesions in the primate Microcebus murinus reveal distinct amyloid beta1-42 and beta1-40 immunoreactivity profiles. AB - Recent reports have shown that amyloid beta deposits in the brains of Alzheimer's disease patients consist mainly of two distinct species of amyloid beta protein (Abeta) with different C-termini, Abeta1-42 (Abeta42) and Abeta1-40 (Abeta40). The nature of the Abeta species in Microcebus murinus brain was investigated immunocytochemically using polyclonal antibodies with clear specificity for the Abeta42 and Abeta40 C-termini. The cortical vascular deposits were immunopositive for both Abeta42 and Abeta40. However, most of the diffuse plaques were strongly positive for Abeta42 whereas only a subset of deposits were positive for Abeta40. Numerous cortical plaques were Abeta42-immunopositive but tested negative for Abeta40. This suggests that Abeta42 is probably associated with early stages of plaque maturation. This neuropathological feature reminiscent of that observed in brains affected by Alzheimer's disease further supports the idea that M. murinus could be used as a potential model of the early stages of this neurological disease. PMID- 10671319 TI - Behavioral disturbances without amyloid deposits in mice overexpressing human amyloid precursor protein with Flemish (A692G) or Dutch (E693Q) mutation. AB - The contribution of mutations in the amyloid precursor protein (APP) gene known as Flemish (APP/A692G) and Dutch (APP/E693Q) to the pathogenesis of Alzheimer's disease and hereditary cerebral hemorrhage with amyloidosis of the Dutch type, respectively, was studied in transgenic mice that overexpress the mutant APP in brain. These transgenic mice showed the same early behavioral disturbances and defects and increased premature death as the APP/London (APP V717I), APP/Swedish (K670N, M671L), and other APP transgenic mice described previously. Pathological changes included intense glial reaction, extensive microspongiosis in the white matter, and apoptotic neurons in select areas of the brain, while amyloid deposits were absent, even in mice over 18 months of age. This contrasts with extensive amyloid deposition in APP/London transgenic mice and less pronounced amyloid deposition in APP/Swedish transgenic mice generated identically. It demonstrated, however, that the behavioral deficiencies and the pathological changes in brain resulting from an impaired neuronal function are caused directly by APP or its proteolytic derivative(s). These accelerate or impinge on the normal process of aging and amyloid deposits per se are not essential for this phenotype. PMID- 10671320 TI - Oxidative insults are associated with apolipoprotein E genotype in Alzheimer's disease brain. AB - The epsilon4 allele of the apolipoprotein E gene (APOE) is associated with sporadic and familial late-onset Alzheimer's disease (AD). Oxidative stress is believed to play an important role in neuronal dysfunction and cell death in AD. We now provide evidence that in the hippocampus of AD, the level of thiobarbituric acid-reactive substances (TBARS) and the APOE genotype are linked. Within AD cases, the levels of TBARS were found to be higher among epsilon4 carriers while the apoE protein concentrations were lower. The relationship between the levels of TBARS and apoE proteins was corroborated by the results from the APOE-deficient mice, in which the levels of TBARS were higher than those in wild-type mice. Among AD cases, tissues from patients with the epsilon4 allele of APOE displayed lower activities of catalase and glutathione peroxidase and lower concentration of glutathione than tissues from patients homozygous for the epsilon3 allele of APOE. Together these data demonstrate that, in AD, the epsilon4 allele of APOE is associated with higher oxidative insults. PMID- 10671321 TI - BDNF blocks caspase-3 activation in neonatal hypoxia-ischemia. AB - Hypoxic-ischemic (H-I) injury to the brain in the perinatal period often leads to significant long-term neurological deficits. In a model of neonatal H-I injury in postnatal day 7 rats, our previous data have shown that cell death with features of apoptosis is prominent between 6 and 24 h after H-I and that neurotrophins, particularly BDNF, can markedly protect against tissue loss. During brain development, caspase-3 is required for normal levels of programmed cell death. Utilizing an antibody specific for the activated form of caspase-3, CM1, we now show that caspase-3 is specifically activated in neuronal cell bodies and their processes beginning at 6 h and peaking 24 h following unilateral carotid ligation and exposure to hypoxia in postnatal day 7 rats. Caspase-3 activation began to occur in cortex at 6 h and in striatum and hippocampus at 12-18 h. Caspase-3 activation was also observed in developing oligodendrocytes. Intracerebroventricular injection of BDNF prior to H-I injury almost completely abolished evidence of H-I-induced caspase-3 activation in vivo. Utilizing a specific molecular marker of an apoptotic pathway, these findings demonstrate that H-I injury to the developing brain is a strong apoptotic stimulus leading to caspase-3 activation, that BDNF can block this process in vivo, and that the ability of BDNF to inhibit caspase activation and subsequent apoptosis likely accounts in large part for its protection against neuronal injury in this model. PMID- 10671322 TI - Enhanced synaptic potentiation in transgenic mice expressing presenilin 1 familial Alzheimer's disease mutation is normalized with a benzodiazepine. AB - Mutations in presenilin 1 (PS1) are the most common causes of familial Alzheimer's disease (FAD). We examined synaptic physiology in hippocampal brain slices of transgenic mice expressing the FAD-linked PS1 deletion of exon 9 variant. Basal excitatory transmission and paired-pulse facilitation in PS1 mutant mice were unchanged. Short- and long-term potentiation of excitatory transmission following high-frequency stimulation were greater in transgenic mice expressing mutant PS1. Mutants had enhanced synaptic inhibition, which may be a compensatory change offsetting an abnormally sensitized plasticity of excitatory transmission. Increasing inhibitory transmission in mutant animals even more with a benzodiazepine reverted synaptic potentiation to the levels of controls. These results support the potential use of benzodiazepines in the treatment of familial Alzheimer's disease. PMID- 10671323 TI - New uses for new and old quinolones and the challenge of resistance. PMID- 10671324 TI - Neutropenia, neutrophil dysfunction, and bacterial infection in patients with human immunodeficiency virus disease: the role of granulocyte colony-stimulating factor. AB - Neutropenia frequently complicates infection due to human immunodeficiency virus (HIV). The etiology of neutropenia in this setting includes bone marrow infection or infiltration, myelosuppressive therapies, the presence of antibodies to HIV, and accelerated apoptosis. Protection against microbial invaders by neutrophils is further compromised by impaired chemotaxis and phagocytosis, production of toxic oxygen species, and expression of cellular adhesion molecules. Neutropenia is a significant risk factor for bacterial infection in HIV-infected patients. Endogenous cytokines, such as granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor, regulate neutrophil count and function. Treatment with recombinant human methionyl G-CSF (filgrastim) has lessened neutropenia in patients with HIV infection. Clinical trials have shown that the incidence of bacterial infections and the number of consequent days of hospitalization for HIV-infected patients receiving filgrastim therapy are lower. Filgrastim treatment also allows administration of larger doses of myelosuppressive agents. PMID- 10671325 TI - Herpes simplex virus type 2 shedding in human immunodeficiency virus-negative men who have sex with men: frequency, patterns, and risk factors. AB - We investigated the frequency, site, and risk factors for herpes simplex virus (HSV) shedding in 30 human immunodeficiency virus (HIV)-negative HSV type 2 (HSV 2)-seropositive men who have sex with men. Subjects collected daily HSV culture samples from genital, perianal, and oral areas for 100 days and maintained diaries of signs and symptoms. Sixteen men (53.3%) shed HSV-2, and 9 (56.3%) of 16 men who were also HSV type 1 (HSV-1)-seropositive shed HSV-1. Overall, HSV-2 was isolated on 3.1% of the days; 68% of the isolations were on days that lesions did not occur. HSV-2 shedding was predominantly perianal (83.3%). HSV-1 was isolated on 2.1% of the days; 23 of 24 HSV-1 isolates were from oral areas. Rates of perianal or genital shedding were 6.6% on the days that participants reported prodromal symptoms and 1.9% on the days that participants did not report prodromal symptoms (P<.001). Men seropositive for both HSV-1 and HSV-2 were significantly more likely to shed HSV-2 (odds ratio, 4.1; 95% confidence interval, 1.4-11.9) than were HSV-2-seropositive men. HSV-2-seropositive men who have sex with men have frequent subclinical HSV-2 shedding, usually from the perianal area, and more frequent prodromal HSV-2 shedding. PMID- 10671326 TI - Asymptomatic herpes simplex virus shedding and Russian roulette. PMID- 10671327 TI - Risk factors for radial artery harvest site infection following coronary artery bypass graft surgery. AB - Radial arteries increasingly are used during coronary artery bypass graft (CABG) surgery. Although risk factors for saphenous vein harvest site infection (HSI) have been reported, rates of and risk factors for radial artery HSI are not well established. We compared rates of radial artery HSI that were detected by 2 surveillance methods, regular and heightened. Risk factors were determined by a case-control study. We identified 35 radial artery HSIs ("case sites") in 26 case patients. The radial artery HSI rate was significantly higher during heightened surveillance than during routine surveillance (12.3% vs. 3.1%, respectively; P=.002). Multivariate analysis showed that diabetes mellitus with a preoperative glucose level >/=200 mg/dL (odds ratio [OR], 4.4; P=. 01) and duration of surgery >/=5 h (OR, 3.1; P=.02) were independent risk factors for radial artery HSI. Infection is a common complication of radial artery harvesting for CABG surgery, and infection rates are dependent on the intensity of surveillance. We identified preoperative hyperglycemia and surgery duration as independent risk factors for radial artery HSI. PMID- 10671328 TI - Invasive disease due to group B Streptococcus in pregnant women and neonates from diverse population groups. AB - From 1993 through 1996, surveillance for invasive disease due to group B Streptococcus (GBS) in neonates aged <7 days and in peripartum pregnant women was performed in a racially and ethnically diverse cohort in 4 cities in the United States. In a birth population of 157,184, 130 neonatal cases (0.8 per 1000) and 54 maternal cases (0.3 per 1000) were identified. Significant correlates with neonatal disease were black or Hispanic race and a birth weight <2500 g. The attack rate for peripartum maternal infection varied widely by city and may have been influenced by the frequency of administration of intrapartum antibiotics or of evaluating febrile women by performance of blood cultures. Pregnancy loss or GBS disease in the infant occurred in 28% of these maternal cases. Among neonatal and maternal GBS isolates, serotypes Ia (34%-37%) and III (25%-26%) predominated, and type V was frequent (14%-23%). These results provide a description of invasive GBS perinatal infection during the period in which guidelines for prevention were actively disseminated. PMID- 10671329 TI - A 5-year review of recurrent group B streptococcal disease: lessons from twin infants. AB - Recurrent invasive disease due to group B Streptococcus (GBS) in twin infants has not been reported. We report 2 cases of recurrent GBS afflicting both siblings of a set of dichorionic twin infants. The maternal and infant colonizing and invasive strains were identical by serotyping and pulsed-field gel electrophoresis (PFGE). Despite attempts at eradication with different antibiotic regimens, the infants remained colonized after treatment of the second episode. A 5-year review of recurrent invasive GBS disease in infants in our affiliated hospitals was undertaken, and 6 further cases were identified. Serotyping and PFGE of isolates from initial and second episodes were genotypically identical for each case. Three infants each had GBS serotype Ia or V disease and 2 had GBS serotype III disease. The exact pathogenesis of recurrent GBS disease remains unclear, but our data support the hypothesis that persistent mucosal colonization with the original GBS strain rather than new acquisition is a pivotal factor in disease recurrence. PMID- 10671330 TI - Early results (at 6 months) with intermittent clarithromycin-including regimens for lung disease due to Mycobacterium avium complex. AB - We initiated a prospective noncomparative trial of treatment for lung disease due to Mycobacterium avium complex (MAC) in human immunodeficiency virus-negative patients, with a regimen of clarithromycin (1000 mg), rifabutin (300-600 mg), and ethambutol (25 mg/kg) administered 3 times per week. Fifty-nine patients were enrolled. Twelve (20%) were lost to follow-up, and 6 (10%) developed clarithromycin intolerance. The remaining 41 patients (69%) completed the initial 6 months of therapy. The sputum of 32 of these patients (78%) converted to negative. When results were compared with the sputum response rates at 6 months in previous studies with a regimen including daily clarithromycin and regimens including intermittent (3 times per week) azithromycin with the same companion drugs, no differences in treatment responses were evident. Adverse reactions related to rifabutin were a major problem, and for 24 (41%) of 59 patients the dosage was decreased or the drug was withdrawn. Intermittent (3 times per week) administration of clarithromycin appears to be as effective as daily administration in effecting sputum conversion in pulmonary MAC disease. PMID- 10671331 TI - A population-based survey of tuberculosis symptoms: how atypical are atypical presentations? AB - There is scant information on tuberculosis symptoms from a population-based perspective. We prospectively identified 526 tuberculosis cases reported in Los Angeles County over a 6-month period. Of 313 persons who completed our questionnaire, 72.7% had cough, 48.2% for >2 weeks, and 52.3% had fever, 29.4% for >2 weeks. Among those with pulmonary disease, only 52.4% had cough for >2 weeks. In a multivariate model, persons with significant symptoms typical of tuberculosis disease (defined as cough or fever for >2 weeks, weight loss, or hemoptysis) were associated with lack of medical insurance, negative tuberculin skin test, diagnosis during a process other than screening, and non-Asian race. In summary, classic symptoms of prolonged cough and fever are insensitive predictors of tuberculosis. Our data suggest that Asians may need to be added to the list of persons who present with tuberculosis atypically. We believe that the Infectious Diseases Society of America guidelines for community-acquired pneumonia should emphasize demographic features in addition to clinical symptoms when suggesting which patients require evaluation for Mycobacterium tuberculosis. PMID- 10671332 TI - A double-blind, randomized, placebo-controlled trial of itraconazole capsules as antifungal prophylaxis for neutropenic patients. AB - To evaluate the efficacy of itraconazole capsules in prophylaxis for fungal infections in neutropenic patients, we conducted a prospective, double-blind, placebo-controlled, randomized trial. Patients with hematologic malignancies or those who received autologous bone marrow transplants were assigned either a regimen of itraconazole (100 mg orally twice daily; n=104) or of placebo (n=106). Overall, fungal infections (superficial or systemic) occurred more frequently in the placebo group (15% vs. 6%; P=.03). There were no differences in the empirical use of amphotericin B or systemic fungal infections. Among patients with neutropenia that was profound (<100 neutrophils/mm3) and prolonged (for at least 7 days), those receiving itraconazole used less empirical amphotericin B (22% vs. 61%; P=.0001) and developed fewer systemic fungal infections (6% vs. 19%; P=.04). For patients with profound and prolonged neutropenia, itraconazole capsules at the dosage of 100 mg every 12 h reduce the frequency of systemic fungal infections and the use of empirical amphotericin B. PMID- 10671333 TI - A prospective search for ocular lesions in hospitalized patients with significant bacteremia. AB - The purpose of this study was to determine the prevalence, risk factors, and prognostic value of ocular lesions in unselected patients with bacteremia. A total of 202 bacteremic patients, 101 nonbacteremic septic patients, and 90 nonseptic control patients were compared in a prospective, controlled, observational study. Ocular lesions related to bacteremia were found in 12% of the bacteremic group, 5% of the septic group, and 2% of the control group. Ocular lesions were significantly more frequent in the bacteremic patients than in the control patients (P=.007). The severity of the clinical condition and the presence of fungemia predict independently a higher risk of ocular lesions. Mortality rates among bacteremic patients with and without ocular lesions were, respectively, 32% and 8% (P<.01; OR, 3.99). The asymptomatic nature of most ocular lesions in patients with bloodstream infections and the impossibility of amelioration in most cases lead us to recommend ophthalmologic examination for bacteremic patients only when prognostic information is needed. PMID- 10671335 TI - An outbreak of Streptococcus pneumoniae serotype 1 in a closed community in southern Israel. AB - An outbreak of Streptococcus pneumoniae serotype 1 occurred in a closed community that was characterized by poverty and crowding. Vaccine was administered to individuals aged >2 years; no new cases occurred among vaccine recipients. Six weeks after vaccination, carriage of serotype 1, but not of other serotypes, decreased 8.8-fold. This suggests that the reduction in serotype 1 carriage reflects the natural course of the outbreak rather than a vaccine effect. Polysaccharide vaccine may be helpful in terminating pneumococcal outbreaks but may not affect pneumococcal carriage. PMID- 10671334 TI - Combination therapy with amprenavir, abacavir, and efavirenz in human immunodeficiency virus (HIV)-infected patients failing a protease-inhibitor regimen: pharmacokinetic drug interactions and antiviral activity. AB - Patients with plasma viral RNA >50,000 copies/mL, despite a protease-inhibitor regimen, received abacavir, amprenavir, and efavirenz to assess efavirenz amprenavir drug interactions and to evaluate safety and antiviral response. Patients first received amprenavir with abacavir and other nucleoside analogs. Amprenavir levels were measured before and after adding efavirenz. Patients then received a second protease inhibitor. There was evidence of genotypic and phenotypic resistance at study entry. No patient had study drugs discontinued because of toxicity. Efavirenz decreased the steady-state area under the curve, maximum plasma concentration, and minimum plasma concentration of amprenavir by 24%, 33%, and 43%, respectively. Three of 10 patients had >1.5 log10 viral response to abacavir and amprenavir. All 8 patients who added efavirenz had >0.5 log10 decline in viral load, and this response lasted >24 weeks for 3 of the patients. A combination regimen that included abacavir, amprenavir, and efavirenz was well tolerated and had sustained activity in some patients. Concomitant efavirenz therapy decreases amprenavir concentrations. PMID- 10671336 TI - Methicillin-resistant Staphylococcus aureus: the other emerging resistant gram positive coccus among liver transplant recipients. AB - We undertook a study of the characteristics and clinical impact of infections due to methicillin-resistant Staphylococcus aureus (MRSA) after liver transplantation. Of 165 patients who received liver transplants at our institution from 1990 through 1998, 38 (23%) developed MRSA infections. The predominant sources of infection were vascular catheters (39%; n=15), wound (18%; n=7), abdomen (18%; n=7), and lung (13%; n=5). A significant increase in MRSA infections (as a percentage of transplant patients infected per year) occurred over time (P=.0001). This increase was greater among intensive care unit patients (P=.001) than among nonintensive care unit hospital patients (P=.17). Cytomegalovirus seronegativity (P=.01) and primary cytomegalovirus infection were significantly associated with MRSA infections (P=.005). Thirty-day mortality among patients with MRSA infections was 21% (8/38). Mortality was 86% in patients with bacteremic MRSA pneumonia or abdominal infection and 6% in those with catheter-related bacteremia (P=.004). Thus the incidence of MRSA infection has increased exponentially among our liver transplant recipients since 1990. These infections have unique risk factors, time of onset, and a significant difference in site-specific mortality; deep-seated bacteremic infections, in particular, portend a grave outcome. PMID- 10671337 TI - Molecular epidemiology of Blastomyces dermatitidis. AB - The inhalation of conidia of Blastomyces dermatitidis, a fungus found in soil, causes disease in humans and animals. We studied the genetic diversity of this pathogen by extracting DNA yeasts and analyzing them with a polymerase chain reaction (PCR)-based typing system we developed, which used restriction fragment analysis of amplicons from the regions between the rDNA repeats and allowed us to class isolates into 3 major groups. Strains were further differentiated by use of PCR fingerprinting with 3 different primers. Fifty-nine isolates collected over 35 years from 15 regions (United States, India, Africa, Canada) were analyzed. Genotypic groups A, B, and C contained 17, 23, and 19 isolates, which were divided into 5, 15, and 12 types, respectively. All 16 isolates from North America in group A were from the upper midwestern United States or Canada, whereas 0 of 20 isolates from the southeastern United States were in group A. Studies of the largest collection from 1 locale (Eagle River, WI), revealed that the soil isolates studied were not responsible for the majority of cases in this outbreak, as previously proposed, and that >1 strain was present in the environment and in patients. Overall, these results provide a tool for the epidemiological study of blastomycosis and illuminate the genetic and geographic diversity of this important pathogen. PMID- 10671338 TI - Bicuspid aortic valve--A silent danger: analysis of 50 cases of infective endocarditis. AB - We analyzed 50 cases of bicuspid aortic valve endocarditis in patients who presented to St. Thomas' Hospital from 1970 through 1998. These represented 12.3% of the 408 cases of native valve endocarditis (NVE). All patients were male, and their mean age was 39 years. Forty-five of the 50 cases were pathologically proven; 47 were clinically definite according to the Duke criteria and 49 according to our modifications of the Duke criteria. Viridans streptococci and staphylococci accounted for 72% of cases. The prevalences of clinical features were similar to those seen in NVE: fever (temperature >/=38 degrees C, 74%) and malaise (70%), although dyspnea was more frequent (36%). There was a high incidence of serious complications (72% heart failure; 30% periannular abscesses). Surgery was required during the initial admission in 82% of cases. Overall mortality was 14%, and surgical mortality was 9%. Few patients knew they had a "heart condition," and a bicuspid aortic valve was detected in only 35% of echocardiograms performed before surgery. PMID- 10671339 TI - Granulocyte-macrophage colony-stimulating factor as immunomodulating factor together with influenza vaccination in stem cell transplant patients. AB - The effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the serological response at influenza vaccination was studied in 117 patients who had undergone stem cell transplantation (SCT). The vaccine response was evaluated as significant increases in levels of influenza hemagglutination-inhibition (HAI) antibodies and of IgG antibodies measured by enzyme-linked immunosorbent assay (ELISA). There was no difference in antibody response to either influenza A or B in 64 patients who received GM-CSF at vaccination, compared with the 53 who did not. In the subgroup of allogeneic SCT patients, HAI showed that the response rate to the influenza B vaccine was significantly higher in the treatment group (P<.05). ELISA showed that autologous SCT patients with breast cancer who received GM-CSF had a better response to influenza A (P<.05) and B (P<.01). At early vaccination, 4-12 months after stem cell transplantation, these responses were more pronounced. GM-CSF appears to improve the response to influenza vaccination in some groups of SCT patients, but only to a limited extent. PMID- 10671340 TI - Hyphal forms in the central nervous system of patients with coccidioidomycosis. AB - Coccidioides immitis is a dimorphic fungus that grows as a filamentous mold in soil and as a spherule at human body temperature. The hyphal or soil form is found rarely in human tissue. We report 5 cases of coccidioidomycosis in which hyphae were found in brain tissue or spinal fluid. The presence of central nervous system plastic devices appears to be associated with morphological reversion to the saprophytic form. This reversion has implications for diagnosis and therapy and may increase the risk of obstruction of the device(s). PMID- 10671341 TI - Nosocomial transmission of Mycobacterium bovis bacille Calmette-Guerin to children receiving cancer therapy and to their health care providers. AB - A previous report of nosocomial infection due to Mycobacterium bovis bacille Calmette-Guerin (BCG) implicated contamination of chemotherapy solutions reconstituted under the same biosafety hood as BCG vaccine used for bladder cancer therapy. We report 3 similar BCG infections in children and describe evidence of respiratory transmission to health care workers (HCWs) from 1 patient. These children were receiving chemotherapy for leukemia when they presented with active tuberculosis. Each isolate was identified biochemically and by both gas-liquid chromatography and major polymorphic tandem repeat-polymerase chain reaction. Pulsed-field gel electrophoresis showed that 2 isolates were identical strains and identical to the Tice and Connaught strains licensed in the United States for bladder chemotherapy. The third isolate differed by a single fragment after DraI restriction. One patient with heavily positive sputum exposed numerous HCWs. Of 41 HCWs, 2 (5%) converted their purified protein derivatives (PPD) skin test. These data underscore the risk of nosocomial BCG transmission by contamination of chemotherapy solutions and demonstrate the potential for transmission to HCWs from patients with active pulmonary disease. PMID- 10671342 TI - Herpes simplex virus DNA in amniotic fluid without neonatal infection. AB - Twenty-one pregnant women were studied to determine the distribution of herpes simplex virus (HSV): 10 had symptomatic genital herpes, including 1 with primary cervical HSV infection, and 11 had asymptomatic genital herpes. Samples from vesicles, the cervix, and amniotic fluid (AF) were analyzed with 2 separate polymerase chain reaction (PCR) tests and with viral culture. For newborns, clinical examinations and pharyngeal HSV cultures were performed, and cord blood IgM antibodies to HSV were measured. HSV DNA was present in the AF of 3 women with symptomatic HSV infection, but all cultures were negative. HSV was detected more often with PCR than with culture, in both vesicle and cervical samples. For the asymptomatic group, all AF samples were negative, whereas 4 cervical samples were positive by PCR (none were positive by culture). All isolates were HSV type 2. All infants were healthy, and none had cord blood IgM antibodies to HSV, including those with PCR-positive AF. PMID- 10671343 TI - Pathogenic significance of methicillin resistance for patients with Staphylococcus aureus bacteremia. AB - To assess whether methicillin resistance is a microbial characteristic associated with deleterious clinical outcome, we performed a cohort study on 908 consecutive episodes of Staphylococcus aureus bacteremia and a case-control study involving 163 pairs of patients matched for preexisting comorbidities, prognosis of the underlying disease, length of hospitalization, and age. Of 908 bacteremic episodes, 225 (24.8%) were due to methicillin-resistant S. aureus (MRSA). Multivariate analysis did not reveal that methicillin resistance was an independent predictor for mortality when shock, source of bacteremia, presence of an ultimately or rapidly fatal underlying disease, acquisition of the infection in an intensive care unit (ICU), inappropriate empirical therapy, female sex, and age were taken into account. Nonetheless, methicillin resistance was an independent predictor for shock. The case-control study could not confirm that shock was linked to MRSA when prior antimicrobial therapy, inappropriate treatment, ICU residence, and female sex were considered. Our data suggest that cohort studies tend to magnify the relationship of MRSA with clinical markers of microbial pathogenicity and that this effect is a shortcoming of these kind of studies that is caused by inadequate control for underlying diseases. PMID- 10671344 TI - Right-side endocarditis in injection drug users: review of proposed mechanisms of pathogenesis. AB - Infective endocarditis of the right-side heart valves occurs commonly in injection drug users. Although a variety of hypotheses have been put forward to explain this clinical observation, no single hypothesis is adequate. In this article, basic scientific, clinical, and microbiological data on this topic are presented. It is apparent that no clear unifying mechanism emerges to explain the well-documented clinical predilection for the infection of the right-side heart valves in this population. Further investigation of this topic utilizing large international clinical registries may help to clarify matters further. PMID- 10671345 TI - Detection of Legionella DNA in peripheral leukocytes, serum, and urine from a patient with pneumonia caused by Legionella dumoffii. AB - The polymerase chain reaction (PCR) has been used to detect Legionella DNA in respiratory tract, serum, and urine samples from patients with pneumonia. In addition, a preliminary study using a guinea pig model suggested that testing of peripheral leukocytes by PCR may be more sensitive than testing of other samples. We used PCR to detect Legionella DNA in serial peripheral leukocyte (buffy coat), serum, and urine samples from a patient with pneumonia caused by Legionella dumoffii. Legionella DNA was detected in all 3 sample types when first collected. Buffy coat and urine samples remained positive up to 56 days after the onset of symptoms, whereas serum samples were positive from 10 up to 16 days after the onset of symptoms. Sequencing of PCR amplicons indicated the presence of L. dumoffii DNA in positive samples. It appears that buffy coat may be a useful sample to test for Legionella DNA, but further study is required to determine the precise sensitivity and to make comparisons with other sample types. PMID- 10671346 TI - Sustained bacteremia associated with transjugular intrahepatic portosystemic shunt (TIPS). AB - Transjugular intrahepatic portosystemic shunt (TIPS) has become a routine procedure in patients with portal hypertension, yet there are few data concerning the incidence of bacteremia associated with this shunt. All patients who underwent TIPS placement at a university hospital from January 1992 through January 1999 were studied. Ninety-nine TIPS were placed, and 10 patients subsequently developed sustained bacteremia; 5 patients had no identifiable source of bacteremia despite rigorous evaluation and were presumed to represent TIPS infections, for an estimated annual incidence of 7 cases/1000 TIPS procedures. Case patients developed bacteremia a median of 100 days after TIPS placement (range, 6-732 days). Bacteremia resolved in all patients after treatment with appropriate intravenous antibiotics (median, 2 weeks of therapy). Although the incidence of TIPS-associated bacteremia appears low, the increasing frequency of this procedure suggests that more information is needed to define this entity and to develop appropriate treatment recommendations. PMID- 10671347 TI - Nose blowing propels nasal fluid into the paranasal sinuses. AB - Intranasal pressures were measured in adults during nose blowing, sneezing, and coughing and were used for fluid dynamic modeling. Sinus CT scans were performed after instillation of radiopaque contrast medium into the nasopharynx followed by nose blowing, sneezing, and coughing. The mean (+/-SD) maximal intranasal pressure was 66 (+/-14) mm Hg during 35 nose blows, 4.6 (+/-3.8) mm Hg during 13 sneezes, and 6.6 (+/-3.8) mm Hg during 18 coughing bouts. A single nose blow can propel up to 1 mL of viscous fluid in the middle meatus into the maxillary sinus. Sneezing and coughing do not generate sufficient pressure to propel viscous fluid into the sinus. Contrast medium from the nasopharynx appeared in >/=1 sinuses in 4 of 4 subjects after a nose blow but not after sneezing or coughing. PMID- 10671348 TI - Residual low-level viral replication could explain discrepancies between viral load and CD4+ cell response in human immunodeficiency virus-infected patients receiving antiretroviral therapy. AB - We report the evolution of chronic infection with human immunodeficiency virus type 1 (HIV-1) in a patient treated with stavudine plus didanosine, whose CD4+ lymphocyte count progressively decreased, despite a sustained plasma viral load <20 copies/mL. After 12 months of therapy, treatment was switched to zidovudine plus lamivudine plus nelfinavir. CD4+ T cell count decreased from 559 x 10(6)/L at month 0 to 259 x 10(6)/L at month 12. Plasma viral load decreased from 21,665 HIV-1 RNA copies/mL at baseline (month 0) to <20 copies/mL after 1 month of therapy with stavudine plus didanosine, and remained below 20 copies/mL until month 12, but always >5 copies/mL. Viral load in tonsilar tissue at month 12 was 125,000 copies/mg of tissue. After the change to triple-drug therapy, the plasma viral load decreased to 5 copies/mL, the CD4+ T cell count increased to 705 x 10(6)/L, and the viral load in tonsilar tissue decreased to <40 copies/mg of tissue at month 24. A low level of HIV-1 replication could explain the lack of immunologic response in patients with apparent virological response. PMID- 10671349 TI - Shiga toxin-producing Escherichia coli urinary tract infection associated with hemolytic-uremic syndrome in an adult and possible adverse effect of ofloxacin therapy. PMID- 10671350 TI - Streptococcus intermedius: A cause of lobar pneumonia with meningitis and brain abscesses. PMID- 10671351 TI - Central nervous system pneumocystosis in AIDS: antemortem diagnosis and successful treatment. PMID- 10671352 TI - Francisella tularensis endocarditis. PMID- 10671354 TI - Cost implications of reporting nonpathogenic protozoa. PMID- 10671353 TI - Trovafloxacin-induced acute hepatitis. PMID- 10671356 TI - Reply PMID- 10671355 TI - Successful treatment of vancomycin-resistant Enterococcus faecium bacteremia with linezolid after failure of treatment with synercid (quinupristin/dalfopristin). PMID- 10671358 TI - Reply PMID- 10671357 TI - Detection of Mycoplasma pneumoniae DNA in cerebrospinal fluid and local immune response. PMID- 10671359 TI - Screening for Chlamydia trachomatis infection in college women with a polymerase chain reaction assay. PMID- 10671360 TI - Factors associated with incomplete virological response to highly active antiretroviral therapy. PMID- 10671361 TI - Rash and opportunistic pneumonia in a malnourished infant adopted from China. PMID- 10671362 TI - Does hepatitis C virus really have no effect on survival in cases of infection with human immunodeficiency virus? PMID- 10671363 TI - Adverse effects of minocycline versus doxycycline in the treatment of Lyme neuroborreliosis. PMID- 10671364 TI - Reply PMID- 10671365 TI - Roles of caspases in apoptosis, development, and cytokine maturation revealed by homozygous gene deficiencies. AB - Caspases are a group of cysteine proteases critical for apoptosis of eukaryotic cells. Deletion of genes that encode murine caspases suggests that caspases are involved not only in apoptosis but also in cytokine maturation and cell growth and differentiation. Among them, caspase-1 and caspase-11 are primarily involved in the processing of pro-inflammatory cytokines. Caspase-3 and caspase-9 are essential for apoptosis during brain development. Caspase-8 is required for the development of heart muscle, cell proliferation in the hematopoietic lineage and death-receptor-mediated apoptosis. These studies suggest that caspases function in cell signaling events including apoptosis, cell growth and differentiation. PMID- 10671366 TI - The actin cytoskeleton of Dictyostelium: a story told by mutants. AB - Actin-binding proteins are effectors of cell signalling and coordinators of cellular behaviour. Research on the Dictyostelium actin cytoskeleton has focused both on the elucidation of the function of bona fide actin-binding proteins as well as on proteins involved in signalling to the cytoskeleton. A major part of this work is concerned with the analysis of Dictyostelium mutants. The results derived from these investigations have added to our understanding of the role of the actin cytoskeleton in growth and development. Furthermore, the studies have identified several cellular and developmental stages that are particularly sensitive to an unbalanced cytoskeleton. In addition, use of GFP fusion proteins is revealing the spatial and temporal dynamics of interactions between actin associated proteins and the cytoskeleton. PMID- 10671367 TI - Epithelial stem cell-mediated development of the human respiratory mucosa in SCID mice. AB - We have developed an in vivo assay for progenitor cells of the human tracheobronchial epithelium relying on the transplantation of human prenatal respiratory tissues into severe combined immunodeficiency mice. Engrafted embryonic or fetal open tracheobronchial rudiments are rapidly closed at each end by a neoformed membrane that we named the operculum. After 2-4 weeks, differentiated human respiratory epithelium covers both the native airway matrix and the new operculum. Human epithelial cells dissociated from either emerging embryonic lung primordia or mature xenografts were seeded in host human airway grafts, of which native epithelium had been eliminated by several cycles of freezing and thawing. All grafts seeded with donor epithelial cells and implanted back into SCID mice recovered a surface mucociliary epithelium expressing expected markers and secreting mucus. Spontaneous epithelium regrowth was never observed in control unseeded, denuded grafts. In some experiments, donor epithelial cells and host denuded airway were sex-mismatched and the donor origin of newly formed epithelial structures was confirmed by sex chromosome detection. After two rounds of seeding and reimplantation, a normal epithelium was observed to line the 3rd generation operculum. These observations substantiate a functional assay for human candidate airway epithelium stem cells. PMID- 10671368 TI - Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nup153 to reforming functional nuclear envelopes. AB - We determined the times when the nuclear membrane, nuclear pore complex (NPC) components, and nuclear import function were recovered during telophase in living HeLa cells. Simultaneous observation of fluorescently-labeled NLS-bearing proteins, lamin B receptor (LBR)-GFP, and Hoechst33342-stained chromosomes revealed that nuclear membranes reassembled around chromosomes by 5 minutes after the onset of anaphase (early telophase) whereas nuclear import function was recovered later, at 8 minutes. GFP-tagged emerin also accumulated on chromosomes 5 minutes after the onset of anaphase. Interestingly, emerin and LBR initially accumulated at distinct, separate locations, but then became uniform 8 minutes after the onset of anaphase, concurrent with the recovery of nuclear import function. We further determined the timing of NPC assembly by immunofluorescence staining of cells fixed at precise times after the onset of anaphase. Taken together, these results showed that emerin, LBR, and several NPC components (RanBP2, Nup153, p62), but not Tpr, reconstitute around chromosomes very early in telophase prior to the recovery of nuclear import activity. PMID- 10671369 TI - Fibronectin fragments induce MMP activity in mouse mammary epithelial cells: evidence for a role in mammary tissue remodeling. AB - Mammary gland form and function are regulated by interactions between epithelium and extracellular matrix. Major glycoprotein components of extracellular matrix have been identified that give survival, proliferation and differentiation signals to mammary epithelial cells. We provide evidence that proteolytic fragments of the extracellular matrix glycoprotein, fibronectin, suppress growth and can promote apoptosis of mouse mammary epithelial cells. During mammary gland involution, total fibronectin and fibronectin fragment levels are increased. The peak levels of fibronectin protein and fragments are observed 4-6 days post weaning, coincident with the peak in epithelial cell death. Using a model for hormone withdrawal-induced death of mammary epithelium, elevated levels of fibronectin proteolytic fragments were associated with apoptosis in TM-6 cells, a tumorigenic mouse mammary epithelial cell line. Treatment of TM-6 cells with exogenous fibronectin fragments (FN120) reduced cell number, and induced apoptosis and matrix degrading protease activity. Inhibition of matrix protease activity rescued TM-6 cell viability, indicating that FN120-induced cell loss is mediated through matrix protease activity. In a three-dimensional model for mammary gland development, FN120 reduced alveolar-like and promoted ductal-like development by a matrix protease-dependent mechanism. These data suggest that during post-lactational involution, fibronectin fragments may contribute to epithelial cell loss and dissolution of mammary alveoli by inducing matrix degrading proteinases. PMID- 10671370 TI - Brain derived versican V2 is a potent inhibitor of axonal growth. AB - In this paper, we identify the chondroitin sulfate proteoglycan versican V2 as a major inhibitor of axonal growth in the extracellular matrix of the mature central nervous system. In immunohistochemical and in situ hybridization experiments we show that this tissue-specific splice variant of versican is predominantly present in myelinated fiber tracts of the brain and in the optic nerve, most likely being expressed by oligodendrocytes. We demonstrate that isolated versican V2 strongly inhibits neurite outgrowth of central and peripheral neurons in stripe-choice assays using laminin-1 as permissive substrate. The inhibitory character of versican V2 is maintained after removal of chondroitin sulfate and N- and O-linked oligosaccharide side chains, but it is abolished after core protein digestion with proteinase-K. Our data support the notion, that intact versican V2 prevents excessive axonal growth during late phases of development and hereby participates in the structural stabilization of the mature central nervous system. PMID- 10671371 TI - Mitotic phosphorylation of SUV39H1, a novel component of active centromeres, coincides with transient accumulation at mammalian centromeres. AB - Centromeres of eukaryotes are frequently associated with constitutive heterochromatin and their activity appears to be coregulated by epigenetic modification of higher order chromatin. Recently, we isolated murine (Suv39h1) and human (SUV39H1) homologues of the dominant Drosophila suppressor of position effect variegation Su(var)3-9, which is also related to the S. pombe silencing factor Clr4. We have shown that mammalian Su(var)3-9 homologues encode novel centromeric proteins on metaphase-arrested chromosomes. Here, we describe a detailed analysis of the chromatin distribution of human SUV39H1 during the cell cycle. Although there is significant heterochromatic overlap between SUV39H1 and M31 (HP1(beta)) during interphase, mitotic SUV39H1 displays a more restricted spatial and temporal association pattern with metaphase chromosomes than M31 (HP1(beta)), or the related HP1(&agr;) gene product. SUV39H1 specifically accumulates at the centromere during prometaphase but dissociates from centromeric positions at the meta- to anaphase transition. In addition, SUV39H1 selectively associates with the active centromere of a dicentric chromosome and also with a neocentromere. Interestingly, SUV39H1 is shown to be a phosphoprotein with modifications at serine and, to a lesser degree, also at threonine residues. Whereas SUV39H1 steady-state protein levels appear constant during the cell cycle, two additional phosphorylated isoforms are detected in mitotic extracts. This intriguing localisation and modification pattern would be consistent with a regulatory role(s) for SUV39H1 in participating in higher order chromatin organisation at mammalian centromeres. PMID- 10671372 TI - Calcium regulation of microtubule sliding in reactivated sea urchin sperm flagella. AB - The changes in the bending pattern of flagella induced by an increased intracellular Ca(2+) concentration are caused by changes in the pattern and velocity of microtubule sliding. However, the mechanism by which Ca(2+) regulates microtubule sliding in flagella has been unclear. To elucidate it, we studied the effects of Ca(2+) on microtubule sliding in reactivated sea urchin sperm flagella that were beating under imposed head vibration. We found that the maximum microtubule sliding velocity obtainable by imposed vibration, which was about 170 180 rad/second in the presence of 250 microM MgATP and <10(-9) M Ca(2+), was decreased by 10(-6)-10(-5) M Ca(2+) by about 15-20%. Similar decrease of the sliding velocity was observed at 54 and 27 microM MgATP. The Ca(2+)-induced decrease of the sliding velocity was due mainly to a decrease in the reverse bend angle. When the plane of beat was artificially rotated by rotating the plane of vibration of the pipette that held the sperm head, the asymmetric bending pattern also rotated at 10(-5) M Ca(2+) as well as at <10(-9) M Ca(2+). The rotation of the bending pattern was observed at MgATP higher than 54 microM ( approximately 100 microM ATP). These results indicate that the Ca(2+)-induced decrease of the sliding velocity is mediated by a rotatable component or components (probably the central pair) at high MgATP, but is not due to specific dynein arms on particular doublets. We further investigated the effects of a mild trypsin treatment and of trifluoperazine on the Ca(2+)-induced decrease in sliding velocity. Axonemes treated for 3 minutes with a low concentration (0.1 microgram/ml) of trypsin beat with a more symmetrical waveform than before the treatment. Also, their microtubule sliding velocity and reverse bend angle were not affected by high Ca(2+) concentrations. Trifluoperazine (25-50 microM) had no effect on the decrease of the sliding velocity in beating flagella at 10(-5) M Ca(2+). However, the flagella that had been 'quiescent' at 10(-4) M Ca(2+) resumed asymmetrical beating following an application of 10-50 microM trifluoperazine. In such beating flagella, both the sliding velocity and the reverse bend angle were close to their respective values at 10(-5) M Ca(2+). Trypsin treatment induced a similar recovery of beating in quiescent flagella at 10(-)(4) M Ca(2+), albeit with a more symmetrical waveform. These results provide first evidence that, at least at ATP concentrations higher than approximately 100 microM, 10(-6)-10(-5) M Ca(2+) decreases the maximum sliding velocity of microtubules in beating flagella through a trypsin-sensitive regulatory mechanism which possibly involves the central pair apparatus. They also suggest that calmodulin may be associated with the mechanism underlying flagellar quiescence induced by 10(-4) M Ca(2+). PMID- 10671373 TI - The large intracytoplasmic loop of the glucose transporter GLUT2 is involved in glucose signaling in hepatic cells. AB - The hypothesis that the glucose transporter GLUT2 can function as a protein mediating transcriptional glucose signaling was addressed. To divert the putative interacting proteins from a glucose signaling pathway, two intracytoplasmic domains of GLUT2, the C terminus and the large loop located between transmembrane domains 6 and 7, were transfected into mhAT3F hepatoma cells. Glucose-induced accumulation of two hepatic gene mRNAs (GLUT2 and L-pyruvate kinase) was specifically inhibited in cells transfected with the GLUT2 loop and not with the GLUT2 C terminus. The dual effects of glucose were dissociated in cells expressing the GLUT2 loop; in fact a normal glucose metabolism into glycogen occurred concomitantly with the inhibition of the glucose-induced transcription. This inhibition by the GLUT2 loop could be due to competitive binding of a protein that normally interacts with endogenous GLUT2. In addition, the GLUT2 loop, tagged with green fluorescent protein (GFP), was located within the nucleus, whereas the GFP and GFP-GLUT2 C-terminal proteins remained in the cytoplasm. In living cells, a fraction (50%) of the expressed GFP-GLUT2 loop translocated rapidly from the cytoplasm to the nucleus in response to high glucose concentration and conversely in the absence of glucose. We conclude that, via protein interactions with its large loop, GLUT2 may transduce a glucose signal from the plasma membrane to the nucleus. PMID- 10671374 TI - Nidogen-1 regulates laminin-1-dependent mammary-specific gene expression. AB - Nidogen-1 (entactin) acts as a bridge between the extracellular matrix molecules laminin-1 and type IV collagen, and thus participates in the assembly of basement membranes. To investigate the role of nidogen-1 in regulating cell-type-specific gene expression in mammary epithelium, we designed a culture microecosystem in which each component, including epithelial cells, mesenchymal cells, lactogenic hormones and extracellular matrix, could be controlled. We found that primary and established mesenchymal and myoepithelial cells synthesized and secreted nidogen 1, whereas expression was absent in primary and established epithelial cells. In an epithelial cell line containing mesenchymal cells, nidogen-1 was produced by the mesenchymal cells but deposited between the epithelial cells. In this mixed culture, mammary epithelial cells express (beta)-casein in the presence of lactogenic hormones. Addition of either laminin-1 plus nidogen-1, or laminin-1 alone, to mammary epithelial cells induced (beta)-casein production. We asked whether recombinant nidogen-1 alone could signal directly for (beta)-casein. Nidogen-1 did not induce (beta)-casein synthesis in epithelial cells, but it augmented the inductive capacity of laminin-1. These data suggest that nidogen-1 can cooperate with laminin-1 to regulate (beta)-casein expression. Addition of full-length nidogen-1 to the mixed cultures had no effect on (beta)-casein gene expression; however, a nidogen-1 fragment containing the laminin-1 binding domain, but lacking the type IV collagen-binding domain, had a dominant negative effect on (beta)-casein expression. These data point to a physiological role for nidogen-1 in the basement membrane-induced gene expression by epithelial cells. PMID- 10671375 TI - Assembly of the exogenous extracellular matrix during basement membrane formation by alveolar epithelial cells in vitro. AB - We found that immortalized alveolar type II epithelial cells (SV40-T2 cells) that were cultured on dense fibrillar collagen supplemented with Matrigel gel formed a thin and continuous lamina densa beneath them. Immunohistochemical analysis of laminin-1, type IV collagen, entactin (nidogen) and perlecan in the culture indicated that all these components were integrated into a sheet structure of basement membrane beneath the cells. Analysis of the temporal and spatial distribution of the basement membrane macromolecules revealed that the initial deposits of laminin-1 and entactin were significantly greater in area in the presence of Matrigel. These globular deposits and the coarse mesh of basement membrane macromolecules developed into a flat membranous basement membrane. In the absence of Matrigel, the SV40-T2 cells failed to form a continuous lamina densa, and the deposits stayed in the coarse mesh. The major biotinylated Matrigel components that were integrated into the basement membrane were laminin 1 and entactin. Furthermore, SV40-T2 cells supplemented with exogenous laminin-1 alone as well as laminin-1 contaminated with entactin formed a continuous lamina densa. These results indicate that the laminin-1 and entactin supplied from the Matrigel were incorporated into a basement membrane beneath the SV40-T2 cells, and contributed to the formation of basement membrane. Therefore, we concluded that the alveolar epithelial cells synthesize laminin-1, entactin, type IV collagen, and perlecan, but that they also needed to assemble exogenous laminin-1 into the basement membrane to complete its formation in vitro. PMID- 10671376 TI - Integrin binding specificity of laminin-10/11: laminin-10/11 are recognized by alpha 3 beta 1, alpha 6 beta 1 and alpha 6 beta 4 integrins. AB - Laminin-10/11, the laminin isoforms containing the alpha 5 chain, are major components of basement membranes of many fetal and adult tissues. Laminin-10/11 purified from the conditioned medium of human lung carcinoma cells were potent in mediating adhesion of the carcinoma cells in an integrin alpha 3 beta 1-dependent manner. To further define the type(s) of integrins involved in cell adhesion to laminin-10/11, we examined the effects of a panel of function-blocking anti integrin antibodies on the adhesion of different cell types to laminin-10/11. Although anti-integrin beta 1 antibody inhibited the adhesion of all cell types tested, anti-alpha 3 antibody inhibited the adhesion of carcinoma and glioma cells but not fibroblastic cells. Adhesion of fibroblastic cells was inhibited, however, by a combination of anti-alpha 3 and anti-alpha 6 antibodies, suggesting that both alpha 3 beta 1 and alpha 6 beta 1 integrins function as laminin-10/11 receptors in these cells. To explore this possibility, we examined the adhesion of K562 leukemic cells transfected with integrin alpha 3 or alpha 6 subunit to laminin-10/11 or other laminin isoforms. Laminin-10/11 were potent adhesive ligands for both the alpha 3 beta 11 and alpha 6 beta 1 transfectants, whereas laminin-5 was the preferred ligand for the alpha 3 beta 1 transfectants. Upon stimulation with the activating anti-integrin beta 1 antibody, both transfectants became more adherent to the substratum regardless of the type of laminins coated, although their preference for laminin isoforms remained unaltered. K562 cells transfected with alpha 6 and beta 4 subunits were also capable of adhering to laminin-10/11, indicating that integrin alpha 6 beta 4 is another receptor for laminin-10/11. Even with lung carcinoma cells, the alpha 6-containing integrins partly contributed to adhesion to laminin-10/11 at higher coating concentrations, although non-integrin receptor(s) might also be involved under such conditions. These results indicated that laminin-10/11 are potent and versatile adhesive ligands in basement membranes capable of binding to both alpha 3 beta 1 and alpha 6 beta 1 integrins with high avidity and also to alpha 6 beta 4 integrin. PMID- 10671377 TI - ARP1 in Golgi organisation and attachment of manchette microtubules to the nucleus during mammalian spermatogenesis. AB - Actin related protein of vertebrate, Arp1, is a major component of the dynactin complex. To characterise and localise Arp1 during mammalian spermatogenesis, polyclonal antibodies were raised against a human recombinant Arp1. Anti-Arp1 antibodies were used for western-immunoblotting, indirect immunofluorescence and immunoelectron microscopy. In round spermatids, Arp1 was detected at the centrosome and at the Golgi apparatus. In elongated spermatids, Arp1 was predominantly found along microtubules of the manchette and at their site of attachment to the nuclear envelope. In maturing spermatids, Arp1 was still present in the pericentriolar material, but in testicular spermatozoa it was not detectable. These various localisations of Arp1 and their changes during spermatid differentiation suggest that the dynactin complex in association with dynein might contribute to several activities: the functional organisation of the centrosome and of the Golgi apparatus and the shaping of the nucleus by manchette microtubules. PMID- 10671378 TI - Selective activation of pre-replication complexes in vitro at specific sites in mammalian nuclei. AB - As the first step in determining whether or not pre-replication complexes are assembled at specific sites along mammalian chromosomes, nuclei from G(1)-phase hamster cells were incubated briefly in Xenopus egg extract in order to initiate DNA replication. Most of the nascent DNA consisted of RNA-primed DNA chains 0.5 to 2 kb in length, and its origins in the DHFR gene region were mapped using both the early labeled fragment assay and the nascent strand abundance assay. The results revealed three important features of mammalian replication origins. First, Xenopus egg extract can selectively activate the same origins of bi directional replication (e.g. ori-beta) and (beta') that are used by hamster cells in vivo. Previous reports of a broad peak of nascent DNA centered at ori (beta/(beta)' appeared to result from the use of aphidicolin to synchronize nuclei and from prolonged exposure of nuclei to egg extracts. Second, these sites were not present until late G(1)-phase of the cell division cycle, and their appearance did not depend on the presence of Xenopus Orc proteins. Therefore, hamster pre-replication complexes appear to be assembled at specific chromosomal sites during G(1)-phase. Third, selective activation of ori-(beta) in late G(1) nuclei depended on the ratio of Xenopus egg extract to nuclei, revealing that epigenetic parameters such as the ratio of initiation factors to DNA substrate could determine the number of origins activated. PMID- 10671379 TI - Import of proteins into the trypanosome nucleus and their distribution at karyokinesis. AB - In all eukaryotic organisms proteins are targeted to the nucleus via a receptor mediated mechanism that requires a specific nuclear localization sequence (NLS) in the protein. Little is known about this process in trypanosomatid protozoa that are considered amongst the earliest divergent eukaryotes. We have used the green fluorescent protein (gfp) and beta-galactosidase reporters to identify the NLS of two trypanosomal proteins, namely the Trypanosoma brucei La protein homologue and histone H2B of T. cruzi. A monopartite NLS was demonstrated at the C terminus of the La protein, whereas a bipartite NLS was identified within the first 40 amino acids of histone H2B. Treatment of live trypanosomes with poisons of ATP synthesis resulted in exit of the La NLS-gfp fusion from the nucleus. Interestingly, this fusion protein accumulated at several discrete sites in the cytoplasm, rather than equilibrating between the nucleus and the cytoplasm. When ATP levels returned to normal, the protein reentered the nucleus, demonstrating that the process was energy dependent. Finally, using fusion proteins that localize to the nucleoplasm or the nucleolus, we identified a subpopulation of mitotic cells in which the chromosomes have segregated but the daughter nuclei remain connected by a thin thread-like structure. We propose that cells containing this structure represent a late stage in nuclear division that can be placed after chromosome segregation, but before completion of karyokinesis. PMID- 10671381 TI - Images in neuroscience. Cognition: procedural memory. PMID- 10671382 TI - How can we learn about developmental processes from cross-sectional studies, or can we? AB - OBJECTIVE: Cross-sectional studies are often used in psychiatric research as a basis of longitudinal inferences about developmental or disease processes. While the limitations of such usage are often acknowledged, these are often understated. The authors describe how such inferences are often, and sometimes seriously, misleading. METHOD: Why and how these inferences mislead are here demonstrated on an intuitive level, by using simulated data inspired by real problems in psychiatric research. RESULTS: Four factors with major roles in the relationship between cross-sectional studies and longitudinal inferences are selection of time scale, type of developmental process studied, reliability of measurement, and clarity of terminology. The authors suggest how to recognize inferential errors when they occur, describe how to protect against such errors in future research, and delineate the circumstances in which only longitudinal studies can answer crucial questions. CONCLUSIONS: The simple conclusion is that one must always use the results of cross-sectional studies to draw inferences about longitudinal processes with trepidation. PMID- 10671383 TI - Late-onset schizophrenia and very-late-onset schizophrenia-like psychosis: an international consensus. The International Late-Onset Schizophrenia Group. AB - OBJECTIVE: Although schizophrenia is generally regarded as an illness with onset in late adolescence or early adult life, a sizeable minority of patients first become ill in middle or old age. Inconsistencies in diagnostic systems and nomenclature, coupled with a tendency among most schizophrenia researchers to ascribe late-onset psychoses to organic factors, have led to such cases occupying an ambiguous position in relation to schizophrenia. Through systematic review of the literature and publication of a consensus statement from an international group of experts in the field, this article aims to clarify the positions of late onset schizophrenia and very-late-onset schizophrenia-like psychosis. METHOD: The authors conducted a MEDLINE literature review and developed a consensus statement summarizing the findings from 2 days of debate and discussion by members of the International Late-Onset Schizophrenia Group. RESULTS: The group achieved consensus on diagnosis, nomenclature, treatment guidelines, and future research directions. CONCLUSIONS: In terms of epidemiology, symptom profile, and identified pathophysiologies, the diagnoses of late-onset schizophrenia (illness onset after 40 years of age) and very-late-onset schizophrenia-like psychosis (onset after 60 years) have face validity and clinical utility. General adoption of these categories will foster systematic investigation of such patients. PMID- 10671384 TI - Risk of recurrence of bipolar disorder in pregnant and nonpregnant women after discontinuing lithium maintenance. AB - OBJECTIVE: Pregnancy poses major challenges for the treatment of bipolar disorder, and information to guide clinical care remains very sparse. The authors sought to determine the illness recurrence risk for women with bipolar disorder who discontinue lithium maintenance during pregnancy. METHOD: The authors retrospectively compared recurrence rates and survival functions for 101 women with DSM-IV bipolar disorder (68 type I, 33 type II) during pregnancy and postpartum (N=42) or during equivalent periods (weeks 1-40 and 41-64) for age matched nonpregnant subjects (N=59) after either rapid (1-14 days) or gradual (15 30 days) discontinuation of lithium. Recurrence rates also were obtained for the year before discontinuing lithium. RESULTS: Rates of recurrence during the first 40 weeks after lithium discontinuation were similar for pregnant (52%) and nonpregnant women (58%) but had been much lower for both in the year before treatment was discontinued (21%). Among subjects who remained stable over the first 40 weeks after lithium discontinuation, postpartum recurrences were 2.9 times more frequent than recurrences in nonpregnant women during weeks 41-64 (70% versus 24%). Depressive or dysphoric-mixed episodes were more prevalent in pregnant than nonpregnant women (63% versus 38% of recurrences). Recurrence risk was greater after rapid than after gradual discontinuation, and for patients with more prior affective episodes, but was similar for diagnostic types I and II. CONCLUSIONS: Rates of recurrence during the first 40 weeks after lithium discontinuation were similar for pregnant and nonpregnant women but then sharply increased postpartum. Risk was much lower during preceding treatment and less with gradual discontinuation. Treatment planning for potentially pregnant women with bipolar disorder should consider the relative risks of fetal exposure to mood stabilizers versus the high recurrence risks after discontinuing lithium. PMID- 10671385 TI - Paroxetine in human breast milk and nursing infants. AB - OBJECTIVE: The purpose of this study was to determine the extent of infant medication exposure through breast-feeding during maternal treatment with paroxetine. METHOD: Breast milk and paired maternal and infant sera were collected after 10 days of maternal treatment with paroxetine at a stable daily dose (10-50 mg/day). All samples were analyzed by means of high-performance liquid chromatography with ultraviolet detection and a limit of detection of 2 ng/ml. RESULTS: Breast milk paroxetine concentrations were highly variable (2-101 ng/ml) and were present in all breast milk samples (N=108). A significant gradient effect was observed, with greater paroxetine concentrations found in later portions of breast milk (hind milk) than in early portions (fore milk). No clear time course of paroxetine excretion into breast milk was demonstrated, although maternal paroxetine daily dose reliably predicted both trough and peak breast milk concentrations over a 24-hour period. In 16 mother and infant serum pairs, no detectable concentrations of paroxetine were found in the serum of the nursing infants. CONCLUSIONS: This study extends previous data by demonstrating the presence of paroxetine in the breast milk of nursing women treated with this medication. The low concentrations of paroxetine in infant serum and lack of any observable adverse effects after maternal use of this medication while breast feeding parallels the available data on other selective serotonin reuptake inhibitors. PMID- 10671386 TI - Further evidence of relation between prenatal famine and major affective disorder. AB - OBJECTIVE: In a previous study, the authors demonstrated an association between prenatal famine in middle to late gestation and major affective disorders requiring hospitalization. In this study, they sought to examine the association by using newly identified cases from the Dutch birth cohort used previously to examine the gender specificity of the association and to assess whether this relation is present for both unipolar and bipolar affective disorders. METHOD: The authors compared the risk of major affective disorder requiring hospitalization in birth cohorts who were and were not exposed, in each trimester of gestation, to famine during the Dutch Hunger Winter of 1944-1945. These cases of major affective disorder requiring hospitalization were newly ascertained from a national psychiatric registry. A larger data set from this registry was used for analysis by gender and diagnostic subtype. RESULTS: For the newly ascertained cases, the risk of developing major affective disorder requiring hospitalization was increased for subjects with exposure to famine in the second trimester and was increased significantly for subjects with exposure in the third trimester, relative to unexposed subjects. For the cases from the entire period of ascertainment, the risk of developing affective disorder was significantly increased for those exposed to famine during the second and the third trimesters of gestation. The effects were demonstrated for men and women and for unipolar and bipolar affective disorders. CONCLUSIONS: These results provide support for the authors' previous findings on the association between middle to late gestational famine and affective disorder. PMID- 10671387 TI - Hypoxic-ischemia-related fetal/neonatal complications and risk of schizophrenia and other nonaffective psychoses: a 19-year longitudinal study. AB - OBJECTIVE: Epidemiologic evidence linking obstetric complications to schizophrenia has been positive but inconclusive. One reason for the lack of conclusive evidence may be the inconsistency in measuring disturbances of fetal/neonatal brain development based on general obstetric markers of maternal health. The authors used data from the National Collaborative Perinatal Project to examine the relationship between schizophrenia and other nonaffective psychoses and a theoretically derived measure of hypoxic-ischemia-related fetal/neonatal complications. METHOD: Six hundred ninety-three men and women (average age 23) born to a community sample of women between 1959 and 1966 were followed up an average of 19 years after early childhood assessments. Subjects with DSM-IV schizophrenia and other nonaffective psychoses were identified using the Diagnostic Interview Schedule and best-estimate consensus diagnoses. RESULTS: Hypoxic-ischemia-related fetal/neonatal complications were associated with a doubling of the risk of developing a psychotic disorder, compared with no relevant complications (6.9% versus 1.4%). When mood disorders were excluded from the group of psychotic diagnoses, the risk of schizophrenia and other nonaffective psychoses associated with hypoxic-ischemia-related fetal/neonatal complications was strikingly elevated, compared with no relevant complications (5.75% versus 0.39%). Nonpsychotic mood disorders were unrelated to these fetal/neonatal complications. Schizophrenia and other nonaffective psychoses were most strongly associated with hypoxic-ischemia-related fetal/neonatal complications of disordered growth and development. CONCLUSIONS: The data show a strikingly elevated, graded, independent risk of schizophrenia and other nonaffective psychoses associated with this classification of antecedent hypoxic ischemia-related fetal/neonatal complications. PMID- 10671388 TI - Relationship of obstetric complications and differences in size of brain structures in monozygotic twin pairs discordant for schizophrenia. AB - OBJECTIVE: The aim of the study was to determine whether a history of obstetric complications and congenital minor physical anomalies are related to differences in the characteristics of brain structures observed within monozygotic twin pairs discordant for schizophrenia. METHOD: The size of the bilateral hippocampi and cerebral ventricles was studied by magnetic resonance imaging in 22 monozygotic twin pairs discordant for schizophrenia. Obstetric complications and minor physical anomalies were independently assessed through parental report and examination, respectively. RESULTS: Compared with the well co-twins, the ill twins consistently had smaller left and right hippocampi as well as larger left lateral ventricles and third ventricles. Relatively small left and right hippocampi were each significantly related to labor-delivery complications and to prolonged labor per se. Relatively large right lateral ventricle size and large total ventricle size were significantly related to labor-delivery complications, prolonged labor, neonatal complications, and total complications for the entire reproductive sequence. In contrast, these brain size differences were not significantly associated with pregnancy complications or minor physical anomalies. CONCLUSIONS: Trauma at the time of labor and delivery and especially prolonged labor appear to be of importance for brain structure anomalies associated with schizophrenia. PMID- 10671389 TI - Phenomenology and outcome of subjects with early- and adult-onset psychotic mania. AB - OBJECTIVE: This study examined clinical differences between subjects with early onset and adult-onset psychotic mania. METHOD: Subjects were from an epidemiologically derived, hospitalized sample who met criteria for definite bipolar disorder after 24 months of follow-up and whose index episode had been manic. Information collected regarding demographic characteristics, psychotic and depressive symptoms, childhood behavior problems and school functioning, substance/alcohol use disorders, and episode recurrence for two subgroups were compared: those whose illness first emerged before age 21 (early onset) (N=23) and those whose first episode occurred after age 30 (adult onset) (N=30). RESULTS: A larger proportion of the early-onset subjects were male, had childhood behavior disorders, had substance abuse comorbidity, exhibited paranoia, and experienced complete episode remission less frequently during 24-month follow-up than the adult-onset subjects. CONCLUSIONS: These data add to the body of evidence that has suggested that many subjects with early-onset psychotic mania have a more severe and developmentally complicated subtype of bipolar disorder. PMID- 10671390 TI - Two-year syndromal and functional recovery in 219 cases of first-episode major affective disorder with psychotic features. AB - OBJECTIVE: Psychotic affective disorders are the most prevalent idiopathic psychoses, but their outcome from onset has rarely been studied. In this study, the authors determined the rate and latency of syndromal recovery and rates of functional recovery after first lifetime hospitalization in patients with first episode psychotic affective disorders. METHOD: From first lifetime hospitalization in 1989-1996, 219 patients with a DSM-IV psychotic affective illness were assessed at intervals over 24 months. Time to syndromal recovery (no longer meeting DSM-IV episode criteria) was assessed by survival analysis, and functional recovery (regaining baseline vocational and residential status) was rated. Factors associated with recovery were identified by bivariate and multivariate methods. RESULTS: By 3, 6, 12, and 24 months after first hospitalization, syndromal recovery was attained by 65.1%, 83.7%, 91.1%, and 97.5%, respectively, of subjects. Time to syndromal recovery (6.1 weeks to 50% of subjects recovered) was shorter for patients who had bipolar disorder, were married, were age 30 or older at onset, lacked comorbidity, required relatively brief hospitalization, and received fewer medicines. Functional recovery by 6 (30.4%) and 24 months (37. 6% of patients) was 2.6-2.7 times less likely than syndromal recovery; 63.1% of those recovering syndromally did not recover functionally by 2 years. Functional recovery was associated with older age at onset and shorter hospitalization. Annual recovery rates remained stable as mean hospital length of stay decreased 3. 6-fold over the 8-year study period. CONCLUSIONS: Syndromal recovery was attained by most psychotic affective disorder patients soon after hospitalization, but only one-third recovered functionally by 24 months. The findings suggest that these very common psychotic illnesses can carry a grave functional prognosis from the initial episode and first hospitalization. PMID- 10671391 TI - Multiple recurrences of major depressive disorder. AB - OBJECTIVE: The authors of this study examined multiple recurrences of unipolar major depressive disorder. METHOD: A total of 318 subjects with unipolar major depressive disorder were prospectively followed for 10 years within a multicenter naturalistic study. Survival analytic techniques were used to examine the probability of recurrence after recovery from the index episode. RESULTS: The mean number of episodes of major depression per year of follow-up was 0. 21, and nearly two-thirds of the subjects suffered at least one recurrence. The number of lifetime episodes of major depression was significantly associated with the probability of recurrence, such that the risk of recurrence increased by 16% with each successive recurrence. The risk of recurrence progressively decreased as the duration of recovery increased. Within subjects, there was very little consistency in the time to recurrence. CONCLUSIONS: Major depressive disorder is a highly recurrent illness. The risk of the recurrence of major depressive disorder progressively increases with each successive episode and decreases as the duration of recovery increases. PMID- 10671392 TI - Lack of seasonal mood change in the Icelandic population: results of a cross sectional study. AB - OBJECTIVE: The prevalence of seasonal affective disorder-as measured by the Seasonal Pattern Assessment Questionnaire-has been found to be unexpectedly low among Icelanders. The aim of this cross-sectional study was to measure seasonal variations in the prevalence of anxiety and depression among Icelanders assessed with the Hospital Anxiety and Depression Questionnaire. METHOD: Four 1, 000 person cohorts, age 20-70 years, selected at random from the Icelandic National Register, were sent the Hospital Anxiety and Depression Scale by mail in either January, April, July, or October. Only responses from the 4-week period after each mailing were considered in the subsequent analysis. RESULTS: The mean anxiety and depression scores in winter were not higher than those in summer for either sex. There was no significant difference between winter and summer in rates of actual or borderline cases of anxiety or depression or for the two categories combined. CONCLUSIONS: This lack of seasonality in anxiety and depression is in sharp contrast to findings from similar cross-sectional studies and may reflect the low propensity for seasonal affective disorder that has been described in the Icelandic population. PMID- 10671393 TI - Female sexual dysfunction associated with antidepressant administration: a randomized, placebo-controlled study of pharmacologic intervention. AB - OBJECTIVE: Few controlled trials of pharmacologic intervention in women with antidepressant-associated sexual dysfunction have been reported, and there is uncertainty about the usefulness of putative treatments and the assessment methodologies. The authors evaluated the efficacy of buspirone and amantadine in the treatment of sexual dysfunction associated with fluoxetine administration. METHOD: Women who had been successfully treated with fluoxetine for at least 8 weeks and who had reported a deterioration in sexual function not present before the initiation of fluoxetine entered a 4-week assessment period. After assessment they were randomly assigned to an 8-week treatment trial with buspirone (N=19), amantadine (N=18), or placebo (N=20). Outcomes were assessed by using a patient rated daily diary and a clinician-rated structured interview. RESULTS: While the amantadine-treated women did report significantly greater improvements in energy levels than women in the placebo group, all treatment groups experienced improvement in overall sexual function as well as in most individual measures. There were no statistically significant differences among the three groups. CONCLUSIONS: Neither buspirone nor amantadine was more effective than placebo in ameliorating antidepressant-associated sexual dysfunction. All groups experienced marked nonspecific improvement during treatment, which suggests the importance of placebo-controlled trials for this condition. PMID- 10671394 TI - Victims of criminal homicide in Sweden: a matched case-control study of health and social risk factors among all 1,739 cases during 1978-1994. AB - OBJECTIVE: The psychiatric and medical characteristics of victims of homicide have not been systematically studied and are often confounded by race. This study was undertaken to determine health and social factors contributing to the risk of being murdered in the Swedish, predominantly Caucasian population. METHOD: All 1,739 homicides between 1978 and 1994 in Sweden were studied in terms of variables in national case registers regarding health, crimes, immigration, and marital status. The same data were extracted for matched comparison persons in the general population, with controls for time of exposure. The data were analyzed by conditional logistic regression on matched pairs. RESULTS: Traumatic brain injury, physical abuse, alcohol dependence, and criminal recidivism conferred risk of being murdered. CONCLUSIONS: To the authors' knowledge, this is the first report of traumatic brain injury, in both men and women, as a risk factor for being murdered. Brain injury may mark risk-taking behavior in general or may cause provocative behavior. PMID- 10671395 TI - Impact of psychiatric conditions on health-related quality of life in persons with HIV infection. AB - OBJECTIVE: Little is known about the impact of comorbid psychiatric symptoms in persons with HIV. This study estimates the burden on health-related quality of life associated with comorbid psychiatric conditions in a nationally representative sample of persons with HIV. METHOD: The authors conducted a multistage sampling of urban and rural areas to produce a national probability sample of persons with HIV receiving medical care in the contiguous United States (N=2,864). Subjects were screened for psychiatric conditions with the short form of the Composite International Diagnostic Interview. Heavy drinking was assessed on the basis of quantity and frequency of drinking. Health-related quality of life was rated with a 28-item instrument adapted from similar measures used in the Medical Outcomes Study. RESULTS: HIV subjects with a probable mood disorder diagnosis had significantly lower scores on health-related quality of life measures than did those without such symptoms. Diminished health-related quality of life was not associated with heavy drinking, and in drug users it was accounted for by presence of a comorbid mood disorder. CONCLUSIONS: Optimization of health-related quality of life is particularly important now that HIV is a chronic disease with the prospect of long-term survival. Comorbid psychiatric conditions may serve as markers for impaired functioning and well-being in persons with HIV. Inclusion of sufficient numbers of appropriately trained mental health professionals to identify and treat such conditions may reduce unnecessary utilization of other health services and improve health-related quality of life in persons with HIV infection. PMID- 10671396 TI - Auditory startle response in trauma survivors with posttraumatic stress disorder: a prospective study. AB - OBJECTIVE: Previous studies have shown elevated autonomic responses to startling tones in trauma survivors with chronic posttraumatic stress disorder (PTSD). The origin of these abnormal responses is obscure. The present study attempted to clarify this issue by prospectively evaluating responses to sudden, loud tones in individuals who arrived at a hospital emergency room after psychologically traumatic events. METHOD: By using a previously established protocol, autonomic and muscular responses to the tones were evaluated at 1 week, 1 month, and 4 months after the traumatic event. Structured diagnostic interviews performed at 4 months classified subjects into groups with (N=36) and without (N=182) PTSD, which were further subdivided according to the presence or absence of major depressive disorder as follows: neither PTSD nor depression (N=166), depression alone (N=16), PTSD alone (N=21), and both PTSD and depression (N=15). RESULTS: The groups showed comparable physiological responses to the tones at 1 week posttrauma. However, at 1 and 4 months posttrauma, the subjects with PTSD showed a greater heart rate response and required more stimulus trials to reach the criteria of skin conductance and orbicularis oculi electromyogram nonresponse. These findings were not significantly influenced by comorbid depression and were not explained by the severity of the traumatic event or by the intensity of the initial symptoms. CONCLUSIONS: Differences in physiological response to startling tones develop along with PTSD in the months that follow a traumatic event. This pattern supports the theories that associate PTSD with progressive neuronal sensitization. PMID- 10671397 TI - Asperger's disorder. PMID- 10671398 TI - Images in psychiatry. Crichton royal hospital. PMID- 10671399 TI - Striatal dopamine transporter binding in neuroleptic-naive patients with schizophrenia studied with positron emission tomography. AB - OBJECTIVE: Recent in vivo imaging studies indicate a dysregulated presynaptic function of the striatal dopaminergic system in patients with schizophrenia. To further explore the basis of this phenomenon, the authors studied brain dopamine transporter binding in vivo in patients with first-episode, never-medicated schizophrenia. METHOD: Nine patients with schizophrenia and nine healthy matched comparison subjects were recruited. Striatal dopamine transporter binding was measured with positron emission tomography and a specific dopamine transporter ligand, [(18)F]CFT, a radiolabeled form of 2beta-carbomethoxy-3beta-(4 fluorophenyl)tropane. RESULTS: Average caudate and putamen dopamine transporter binding potentials were almost identical in the patients and comparison subjects, but the patients lacked the right-left asymmetry of the caudate dopamine transporter binding seen in the comparison group. CONCLUSIONS: Average striatal dopamine transporter density is unaltered in neuroleptic-naive patients with schizophrenia. However, patients lack asymmetry in caudate dopamine transporter binding, which conforms with disrupted brain lateralization in this disorder. PMID- 10671400 TI - Genetic influence on laterality in schizophrenia? A twin study of neurological soft signs. AB - OBJECTIVE: This study explored the genetic basis of neurological soft signs in schizophrenia and addressed disturbed hemispheric lateralization. METHOD: The authors investigated neurological soft signs in 30 monozygotic twin pairs, 13 pairs discordant for schizophrenia or schizoaffective disorder and 17 healthy comparison twin pairs. RESULTS: The twins with schizophrenia showed higher total scores for neurological soft signs than did the comparison subjects. The total scores for neurological soft signs of the nonaffected discordant twins were significantly higher than those of the comparison twins. There was a significant difference between the nonaffected and affected discordant twins in total scores for neurological soft signs. In contrast to the comparison subjects, the nonaffected and affected twins of the discordant pairs showed a trend toward higher scores for neurological soft signs on the left body half. CONCLUSIONS: These results suggest that the occurrence of neurological soft signs and, more specifically, their lateralization to the left body half are genetically transmitted. PMID- 10671401 TI - Verbal working memory impairment in schizophrenia patients and their first-degree relatives: evidence from the digit span task. AB - OBJECTIVE: The evidence for verbal working memory deficits in schizophrenia has been inconsistent. Few studies have evaluated verbal working memory in the first degree relatives of schizophrenia patients, who likely share the genetic diathesis for schizophrenia but not the potential confounds associated with chronic mental illness. METHOD: The Wechsler Digit Span Task was used to investigate verbal working memory in 52 schizophrenia patients, 56 of their first degree relatives, and 73 nonpsychiatric comparison subjects. RESULTS: The nonpsychotic relatives showed no impairment on the forward digit span task, a measure of general attention, but did show impairment on the backward digit span task, a measure of verbal working memory. Schizophrenia patients showed impairment on both the forward and backward digit span tasks. CONCLUSIONS: These results indicate that the forward and backward digit span tasks tap different cognitive abilities that are differentially associated with the diathesis for schizophrenia. Working memory deficits associated with schizophrenia appear to be generalized and not limited to the spatial modality. PMID- 10671402 TI - Alterations in the functional anatomy of working memory in adult attention deficit hyperactivity disorder. AB - OBJECTIVE: The authors used a functional neuroimaging study with a working memory probe to investigate the pathophysiology of attention deficit hyperactivity disorder (ADHD). Their goal was to compare regional cerebral blood flow (rCBF) changes related to working memory in adults with and without ADHD. METHOD: Using [(15)O]H(2)O positron emission tomography (PET) studies, the authors compared the sites of neural activation related to working memory in six adult men diagnosed with ADHD and six healthy men without ADHD who were matched in age and general intelligence. RESULTS: Task-related changes in rCBF in the men without ADHD were more prominent in the frontal and temporal regions, but rCBF changes in men with ADHD were more widespread and primarily located in the occipital regions. CONCLUSIONS: These data suggest the use of compensatory mental and neural strategies by subjects with ADHD in response to a disrupted ability to inhibit attention to nonrelevant stimuli and the use of internalized speech to guide behavior. PMID- 10671403 TI - MRI assessment of children with obsessive-compulsive disorder or tics associated with streptococcal infection. AB - OBJECTIVE: The authors assessed selective basal ganglia involvement in a subgroup of children with obsessive-compulsive disorder (OCD) and/or tics believed to be associated with streptococcal infection. METHOD: Using computer-assisted morphometric techniques, they analyzed the cerebral magnetic resonance images of 34 children with presumed streptococcus-associated OCD and/or tics and 82 healthy comparison children who were matched for age and sex. RESULTS: The average sizes of the caudate, putamen, and globus pallidus, but not of the thalamus or total cerebrum, were significantly greater in the group of children with streptococcus associated OCD and/or tics than in the healthy children. The differences were similar to those found previously for subjects with Sydenham's chorea compared with normal subjects. CONCLUSIONS: These results support the hypothesis that there is a distinct subgroup of subjects with OCD and/or tics who have enlarged basal ganglia. These findings are consistent with the hypothesis of an autoimmune response to streptococcal infection. PMID- 10671404 TI - Altered cAMP-dependent protein kinase A in platelets of patients with obsessive compulsive disorder. AB - OBJECTIVE: The purpose of this study was to assess cAMP-dependent protein kinase A in patients with obsessive-compulsive disorder (OCD). METHOD: The levels and the activity of protein kinase A were evaluated in whole platelets obtained from 12 unmedicated patients with OCD and 15 healthy comparison subjects. RESULTS: The immunolabeling of protein kinase A regulatory subunits type I and II were significantly greater but that of the catalytic subunit significantly lower in patients with OCD than in healthy subjects. The cAMP-stimulated activity in patients with OCD was significantly lower than that in healthy subjects. CONCLUSIONS: These data suggest a possible role of protein kinase A in the pathophysiology of OCD. PMID- 10671405 TI - Use of the selective serotonin 3 receptor antagonist ondansetron in the treatment of neuroleptic-induced tardive dyskinesia. AB - OBJECTIVE: The authors examined the efficacy, tolerability, and safety of ondansetron, a selective serotonin 3 receptor antagonist, in patients with tardive dyskinesia. METHOD: Twenty patients with schizophrenia who had neuroleptic-induced tardive dyskinesia were given 12 mg/day of ondansetron for 12 weeks in an open-label study. RESULTS: Administration of ondansetron resulted in a statistically significant improvement in tardive dyskinesia and psychotic symptoms. CONCLUSIONS: Ondansetron may be an effective and safe therapy to control tardive dyskinesia and psychosis in patients with schizophrenia. PMID- 10671406 TI - Prediction of detached personality in healthy subjects by low dopamine transporter binding. AB - OBJECTIVE: Low striatal dopamine D(2) receptor binding in healthy human subjects has been associated with detached personality in studies using positron emission tomography (PET) and the Karolinska Scales of Personality questionnaire. The authors investigated whether a similar correlation exists between striatal dopamine transporter binding and detached personality. METHOD: Eighteen healthy volunteers participated in a PET study with the specific dopamine transporter ligand [(18)F]CFT ([(18)F]WIN 35,428) and completed the Karolinska Scales of Personality questionnaire form. RESULTS: Age-corrected dopamine transporter binding in the putamen, but not in the caudate, correlated negatively with detachment personality scores, especially in the right hemisphere. CONCLUSIONS: This finding supports the hypothesis that low dopaminergic neurotransmission is associated with detached personality. Furthermore, since [(18)F]CFT binding is thought to reflect the density of dopaminergic nerve terminals in the brain, the authors suggest that the neurodevelopmental formation of the brain dopaminergic system may influence adult personality traits. PMID- 10671419 TI - Bright light therapy's effect on postpartum depression. PMID- 10671420 TI - Risperidone treatment for psychosis in end-stage Friedreich's ataxia. PMID- 10671421 TI - Intoxication with olanzapine. PMID- 10671422 TI - Dextromethorphan-induced psychosis. PMID- 10671424 TI - Cost-effectiveness of psychiatrists. PMID- 10671425 TI - Conflicted caregivers. PMID- 10671427 TI - Magnetic resonance imaging abnormalities and psychiatric illness. PMID- 10671428 TI - Cost-effectiveness of psychiatrists. PMID- 10671429 TI - Cognitive effects of testosterone supplementation. PMID- 10671431 TI - Theoretical-clinical-empirical approach to classifying axis II disorders. PMID- 10671434 TI - Sympathoadrenal hyperactivity and neuroleptic malignant syndrome. PMID- 10671435 TI - Comparison of clozapine and risperidone. PMID- 10671437 TI - Cyanobacterial cell walls: news from an unusual prokaryotic envelope. PMID- 10671438 TI - Role of the H protein in assembly of the photochemical reaction center and intracytoplasmic membrane in Rhodospirillum rubrum. AB - Rhodospirillum rubrum is a model for the study of membrane formation. Under conditions of oxygen limitation, this facultatively phototrophic bacterium forms an intracytoplasmic membrane that houses the photochemical apparatus. This apparatus consists of two pigment-protein complexes, the light-harvesting antenna (LH) and photochemical reaction center (RC). The proteins of the photochemical components are encoded by the puf operon (LHalpha, LHbeta, RC-L, and RC-M) and by puhA (RC-H). R. rubrum puf interposon mutants do not form intracytoplasmic membranes and are phototrophically incompetent. The puh region was cloned, and DNA sequence determination identified open reading frames bchL and bchM and part of bchH; bchHLM encode enzymes of bacteriochlorophyll biosynthesis. A puhA/G115 interposon mutant was constructed and found to be incapable of phototrophic growth and impaired in intracytoplasmic membrane formation. Comparison of properties of the wild-type and the mutated and complemented strains suggests a model for membrane protein assembly. This model proposes that RC-H is required as a foundation protein for assembly of the RC and highly developed intracytoplasmic membrane. In complemented strains, expression of puh occurred under semiaerobic conditions, thus providing the basis for the development of an expression vector. The puhA gene alone was sufficient to restore phototrophic growth provided that recombination occurred. PMID- 10671439 TI - Enhanced nitrogenase activity in strains of Rhodobacter capsulatus that overexpress the rnf genes. AB - In the photosynthetic bacterium Rhodobacter capsulatus, a putative membrane-bound complex encoded by the rnfABCDGEH operon is thought to be dedicated to electron transport to nitrogenase. In this study, the whole rnf operon was cloned under the control of the nifH promoter in plasmid pNR117 and expressed in several rnf mutants. Complementation analysis demonstrated that transconjugants which integrated plasmid pNR117 directed effective biosynthesis of a functionally competent complex in R. capsulatus. Moreover, it was found that strains carrying pNR117 displayed nitrogenase activities 50 to 100% higher than the wild-type level. The results of radioactive labeling experiments indicated that the intracellular content of nitrogenase polypeptides was marginally altered in strains containing pNR117, whereas the levels of the RnfB and RnfC proteins present in the membrane were four- and twofold, respectively, higher than the wild-type level. Hence, the enhancement of in vivo nitrogenase activity was correlated with a commensurate overproduction of the Rnf polypeptides. In vitro nitrogenase assays performed in the presence of an artificial electron donor indicated that the catalytic activity of the enzyme was not increased in strains overproducing the Rnf polypeptides. It is proposed that the supply of reductants through the Rnf complex might be rate limiting for nitrogenase activity in vivo. Immunoprecipitation experiments performed on solubilized membrane proteins revealed that RnfB and RnfC are associated with each other and with additional polypeptides which may be components of the membrane-bound complex. PMID- 10671440 TI - Autoinduction of 2,4-diacetylphloroglucinol biosynthesis in the biocontrol agent Pseudomonas fluorescens CHA0 and repression by the bacterial metabolites salicylate and pyoluteorin. AB - The antimicrobial metabolite 2,4-diacetylphloroglucinol (2,4-DAPG) contributes to the capacity of Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soilborne pathogens. A 2, 4-DAPG-negative Tn5 insertion mutant of strain CHA0 was isolated, and the nucleotide sequence of the 4-kb genomic DNA region adjacent to the Tn5 insertion site was determined. Four open reading frames were identified, two of which were homologous to phlA, the first gene of the 2,4-DAPG biosynthetic operon, and to the phlF gene encoding a pathway specific transcriptional repressor. The Tn5 insertion was located in an open reading frame, tentatively named phlH, which is not related to known phl genes. In wild-type CHA0, 2, 4-DAPG production paralleled expression of a phlA'-'lacZ translational fusion, reaching a maximum in the late exponential growth phase. Thereafter, the compound appeared to be degraded to monoacetylphloroglucinol by the bacterium. 2,4-DAPG was identified as the active compound in extracts from culture supernatants of strain CHA0 specifically inducing phlA'-'lacZ expression about sixfold during exponential growth. Induction by exogenous 2,4-DAPG was most conspicuous in a phlA mutant, which was unable to produce 2, 4-DAPG. In a phlF mutant, 2,4-DAPG production was enhanced severalfold and phlA'-'lacZ was expressed at a level corresponding to that in the wild type with 2,4-DAPG added. The phlF mutant was insensitive to 2,4-DAPG addition. A transcriptional phlA-lacZ fusion was used to demonstrate that the repressor PhlF acts at the level of transcription. Expression of phlA'-'lacZ and 2,4-DAPG synthesis in strain CHA0 was strongly repressed by the bacterial extracellular metabolites salicylate and pyoluteorin as well as by fusaric acid, a toxin produced by the pythopathogenic fungus Fusarium. In the phlF mutant, these compounds did not affect phlA'-'lacZ expression and 2, 4-DAPG production. PhlF-mediated induction by 2,4-DAPG and repression by salicylate of phlA'-'lacZ expression was confirmed by using Escherichia coli as a heterologous host. In conclusion, our results show that autoinduction of 2,4-DAPG biosynthesis can be countered by certain bacterial (and fungal) metabolites. This mechanism, which depends on phlF function, may help P. fluorescens to produce homeostatically balanced amounts of extracellular metabolites. PMID- 10671441 TI - Expression of a new operon from Bacillus subtilis, ykzB-ykoL, under the control of the TnrA and PhoP-phoR global regulators. AB - The ykzB and ykoL genes encode two peptides, of 51 and 60 amino acids, the functions of which are unknown. The ykzB and tnrA genes are contiguous and transcribed divergently. Expression of ykzB and ykoL is induced by glutamate and is under the control of the TnrA global regulator of nitrogen utilization. TnrA regulated its own synthesis in glutamate minimal medium. Two DNA sequences (TnrAB1 and TnrAB2) homologous to the TnrA binding site are present in the region between tnrA and ykzB. Deletion mapping indicated that the TnrAB2 binding site was involved in activation of the ykzB promoter. In addition, transcription of tnrA depends on the presence of the TnrAB1 binding site. The ykzB and ykoL genes are probably in the same transcriptional unit. A single promoter involved in transcription in the presence of glutamate was mapped by primer extension. ykoL expression was induced by phosphate limitation and depended on the PhoP-PhoR two component regulatory system. Its promoter was mapped to the region between ykoL and ykzB. Four boxes similar to the PhoP binding site are present upstream from the ykoL promoter. These boxes are probably recognized by PhoP approximately P during the activation of transcription in phosphate limitation conditions. PMID- 10671442 TI - Characterization and role of tbuX in utilization of toluene by Ralstonia pickettii PKO1. AB - The tbu regulon of Ralstonia pickettii PKO1 encodes enzymes involved in the catabolism of toluene, benzene, and related alkylaromatic hydrocarbons. The first operon in this regulon contains genes that encode the tbu pathway's initial catabolic enzyme, toluene-3-monooxygenase, as well as TbuT, the NtrC-like transcriptional activator for the entire regulon. It has been previously shown that the organization of tbuT, which is located immediately downstream of tbuA1UBVA2C, and the associated promoter (PtbuA1) is unique in that it results in a cascade type of up-regulation of tbuT in response to a variety of effector compounds. In our efforts to further characterize this unusual mode of gene regulation, we discovered another open reading frame, encoded on the strand opposite that of tbuT, 63 bp downstream of the tbuT stop codon. The 1,374-bp open reading frame, encoding a 458-amino-acid peptide, was designated tbuX. The predicted amino acid sequence of TbuX exhibited significant similarity to several putative outer membrane proteins from aromatic hydrocarbon-degrading bacteria, as well as to FadL, an outer membrane protein needed for uptake of long-chain fatty acids in Escherichia coli. Based on sequence analysis, transcriptional and expression studies, and deletion analysis, TbuX seems to play an important role in the catabolism of toluene in R. pickettii PKO1. In addition, the expression of tbuX appears to be regulated in a manner such that low levels of TbuX are always present within the cell, whereas upon toluene exposure these levels dramatically increase, even more than those of toluene-3-monooxygenase. This expression pattern may relate to the possible role of TbuX as a facilitator of toluene entry into the cell. PMID- 10671443 TI - KorSA from the Streptomyces integrative element pSAM2 is a central transcriptional repressor: target genes and binding sites. AB - pSAM2, a 10.9-kb mobile integrative genetic element from Streptomyces ambofaciens, possesses, as do a majority of Streptomyces conjugative plasmids, a kil-kor system associated with its transfer. The kor function of pSAM2 was attributed to the korSA gene, but its direct role remained unclear. The present study was focused on the determination of the KorSA targets. It was shown that KorSA acts as a transcriptional repressor by binding to a conserved 17-nucleotide sequence found upstream of only two genes: its own gene, korSA, and pra, a gene positively controlling pSAM2 replication, integration, and excision. A unique feature of KorSA, compared to Kor proteins from other Streptomyces conjugative plasmids, is that it does not directly regulate pSAM2 transfer. KorSA does not bind to the pSAM2 genes coding for transfer and intramycelial spreading. Through the repression of pra, KorSA is able to negatively regulate pSAM2 functions activated by Pra and, consequently, to maintain pSAM2 integrated in the chromosome. PMID- 10671444 TI - Regulation of cold shock-induced RNA helicase gene expression in the Cyanobacterium anabaena sp. strain PCC 7120. AB - Expression of the Anabaena sp. strain PCC 7120 RNA helicase gene crhC is induced by cold shock. crhC transcripts are not detectable at 30 degrees C but accumulate at 20 degrees C, and levels remain elevated for the duration of the cold stress. Light-derived metabolic capability, and not light per se, is required for crhC transcript accumulation. Enhanced crhC mRNA stability contributes significantly to the accumulation of crhC transcripts, with the crhC half-life increasing sixfold at 20 degrees C. The accumulation is reversible, with the cells responding more rapidly to temperature downshifts than to upshifts, as a result of the lack of active mRNA destabilization and the continuation of crhC transcription, at least transiently, after a temperature upshift. Translational inhibitors do not induce crhC expression to cold shock levels, indicating that inhibition of translation is only one of the signals required to activate the cold shock response in Anabaena. Limited amounts of protein synthesis are required for the cold shock-induced accumulation of crhC transcripts, as normal levels of accumulation occur in the presence of tetracycline but are abolished by chloramphenicol. Regulation of crhC expression may also extend to the translational level, as CrhC protein levels do not correlate completely with the pattern of mRNA transcript accumulation. Our experiments indicate that the regulation of crhC transcript accumulation is tightly controlled by both temperature and metabolic activity at the levels of transcription, mRNA stabilization, and translation. PMID- 10671445 TI - Identification of a chitin-binding protein secreted by Pseudomonas aeruginosa. AB - One of the major proteins secreted by Pseudomonas aeruginosa is a 43-kDa protein, which is cleaved by elastase into smaller fragments, including a 30-kDa and a 23 kDa fragment. The N-terminal 23-kDa fragment was previously suggested as corresponding to a staphylolytic protease and was designated LasD (S. Park and D. R. Galloway, Mol. Microbiol. 16:263-270, 1995). However, the sequence of the gene encoding this 43-kDa protein revealed that the N-terminal half of the protein is homologous to the chitin-binding proteins CHB1 of Streptomyces olivaceoviridis and CBP21 of Serratia marcescens and to the cellulose-binding protein p40 of Streptomyces halstedii. Furthermore, a short C-terminal fragment shows homology to a part of chitinase A of Vibrio harveyi. The full-length 43-kDa protein could bind chitin and was thereby protected against the proteolytic activity of elastase, whereas the degradation products did not bind chitin. The purified 43 kDa chitin-binding protein had no staphylolytic activity, and comparison of the enzymatic activities in the extracellular medium of a wild-type strain and a chitin-binding protein-deficient mutant indicated that the 43-kDa protein supports neither chitinolytic nor staphylolytic activity. We conclude that the 43 kDa protein, which was found to be produced by many clinical isolates of P. aeruginosa, is a chitin-binding protein, and we propose to name it CbpD (chitin binding protein D). PMID- 10671446 TI - FabF is required for piezoregulation of cis-vaccenic acid levels and piezophilic growth of the deep-Sea bacterium Photobacterium profundum strain SS9. AB - To more fully explore the role of unsaturated fatty acids in high-pressure, low temperature growth, the fabF gene from the psychrotolerant, piezophilic deep-sea bacterium Photobacterium profundum strain SS9 was characterized and its role and regulation were examined. An SS9 strain harboring a disruption in the fabF gene (strain EA40) displayed growth impairment at elevated hydrostatic pressure concomitant with diminished cis-vaccenic acid (18:1) production. However, growth ability at elevated pressure could be restored to wild-type levels by the addition of exogenous 18:1 to the growth medium. Transcript analysis did not indicate that the SS9 fabF gene is transcriptionally regulated, suggesting that the elevated 18:1 levels produced in response to pressure increase result from posttranscriptional changes. Unlike many pressure-adapted bacterial species such as SS9, the mesophile Escherichia coli did not regulate its fatty acid composition in an adaptive manner in response to changes in hydrostatic pressure. Moreover, an E. coli fabF strain was as susceptible to elevated pressure as wild type cells. It is proposed that the SS9 fabF product, beta-ketoacyl-acyl carrier protein synthase II has evolved novel pressure-responsive characteristics which facilitate SS9 growth at high pressure. PMID- 10671447 TI - Characterization of a thermostable DNA glycosylase specific for U/G and T/G mismatches from the hyperthermophilic archaeon Pyrobaculum aerophilum. AB - U/G and T/G mismatches commonly occur due to spontaneous deamination of cytosine and 5-methylcytosine in double-stranded DNA. This mutagenic effect is particularly strong for extreme thermophiles, since the spontaneous deamination reaction is much enhanced at high temperature. Previously, a U/G and T/G mismatch specific glycosylase (Mth-MIG) was found on a cryptic plasmid of the archaeon Methanobacterium thermoautotrophicum, a thermophile with an optimal growth temperature of 65 degrees C. We report characterization of a putative DNA glycosylase from the hyperthermophilic archaeon Pyrobaculum aerophilum, whose optimal growth temperature is 100 degrees C. The open reading frame was first identified through a genome sequencing project in our laboratory. The predicted product of 230 amino acids shares significant sequence homology to [4Fe-4S] containing Nth/MutY DNA glycosylases. The histidine-tagged recombinant protein was expressed in Escherichia coli and purified. It is thermostable and displays DNA glycosylase activities specific to U/G and T/G mismatches with an uncoupled AP lyase activity. It also processes U/7,8-dihydro-oxoguanine and T/7,8-dihydro oxoguanine mismatches. We designate it Pa-MIG. Using sequence comparisons among complete bacterial and archaeal genomes, we have uncovered a putative MIG protein from another hyperthermophilic archaeon, Aeropyrum pernix. The unique conserved amino acid motifs of MIG proteins are proposed to distinguish MIG proteins from the closely related Nth/MutY DNA glycosylases. PMID- 10671448 TI - Autophosphorylation of phosphoglucosamine mutase from Escherichia coli. AB - Phosphoglucosamine mutase (GlmM) catalyzes the formation of glucosamine-1 phosphate from glucosamine-6-phosphate, an essential step in the pathway for UDP N-acetylglucosamine biosynthesis in bacteria. This enzyme must be phosphorylated to be active and acts according to a ping-pong mechanism involving glucosamine-1, 6-diphosphate as an intermediate (L. Jolly, P. Ferrari, D. Blanot, J. van Heijenoort, F. Fassy, and D. Mengin-Lecreulx, Eur. J. Biochem. 262:202-210, 1999). However, the process by which the initial phosphorylation of the enzyme is achieved in vivo remains unknown. Here we show that the phosphoglucosamine mutase from Escherichia coli autophosphorylates in vitro in the presence of [(32)P]ATP. The same is observed with phosphoglucosamine mutases from other bacterial species, yeast N-acetylglucosamine-phosphate mutase, and rabbit muscle phosphoglucomutase. Labeling of the E. coli GlmM enzyme with [(32)P]ATP requires the presence of a divalent cation, and the label is subsequently lost when the enzyme is incubated with either of its substrates. Analysis of enzyme phosphorylation by high-pressure liquid chromatography and coupled mass spectrometry confirms that only one phosphate has been covalently linked to the enzyme. Only phosphoserine could be detected after acid hydrolysis of the labeled protein, and site-directed mutagenesis of serine residues located in or near the active site identifies the serine residue at position 102 as the site of autophosphorylation of E. coli GlmM. PMID- 10671449 TI - WhiD and WhiB, homologous proteins required for different stages of sporulation in Streptomyces coelicolor A3(2). AB - The whiD locus, which is required for the differentiation of Streptomyces coelicolor aerial hyphae into mature spore chains, was localized by map-based cloning to the overlap between cosmids 6G4 and D63 of the minimal ordered library of Redenbach et al. (M. Redenbach et al., Mol. Microbiol. 21:77-96, 1996). Subcloning and sequencing showed that whiD encodes a homologue of WhiB, a protein required for the initiation of sporulation septation in S. coelicolor. WhiD and WhiB belong to a growing family of small (76- to 112-residue) proteins of unknown biochemical function in which four cysteines are absolutely conserved; all known members of this family are found in the actinomycetes. A constructed whiD null mutant showed reduced levels of sporulation, and those spores that did form were heat sensitive, lysed extensively, and were highly irregular in size, arising at least in part from irregularity in septum placement. The whiD null mutant showed extreme variation in spore cell wall deposition; most spores had uniformly thin (20- to 30-nm) walls, but spore chains were frequently observed in which there was irregular but very pronounced (up to 170 nm) cell wall thickening at the junctions between spores. whiD null mutant spores were frequently partitioned into irregular smaller units through the deposition of additional septa, which were often laid down in several different planes, very close to the spore poles. These "minicompartments" appeared to be devoid of chromosomal DNA. Two whiD promoters, whiDp1 and whiDp2, were identified, and their activities were analyzed during development of wild-type S. coelicolor on solid medium. Both promoters were developmentally regulated; whiDp1 and whiDp2 transcripts were detected transiently, approximately at the time when sporulation septa were observed in the aerial hyphae. PMID- 10671450 TI - Differential distribution of novel restriction-modification systems in clonal lineages of Neisseria meningitidis. AB - Using representational difference analysis, we isolated novel meningococcal restriction-modification (R-M) systems. NmeBI, which is a homologue of the R-M system HgaI of Pasteurella volantium, was present in meningococci of the ET-5 complex and of lineage III. NmeAI was found in serogroup A, ET-37 complex, and cluster A4 meningococci. NmeDI was harbored by meningococci of the ET-37 complex and of cluster A4, but not by serogroup A meningococci. Two of the R-M systems, NmeBI and NmeDI, were located at homologous positions between the phenylalanyl tRNA synthetase genes pheS and pheT, which appeared to be a preferential target for the insertion of foreign DNA in meningococci. The distribution of the three R M systems was tested with 103 meningococcal strains comprising 49 sequence types. The vast majority of the strains had either NmeBI, NmeAI, or both NmeAI and NmeDI. Using cocultivation experiments, we could demonstrate that NmeBI, which was present in ET-5 complex meningococci, was responsible for a partial restriction of DNA transfer from meningococci of the ET-37 complex to meningococci of the ET-5 complex. PMID- 10671451 TI - Extracellular glycanases of Rhizobium leguminosarum are activated on the cell surface by an exopolysaccharide-related component. AB - Rhizobium leguminosarum secretes two extracellular glycanases, PlyA and PlyB, that can degrade exopolysaccharide (EPS) and carboxymethyl cellulose (CMC), which is used as a model substrate of plant cell wall cellulose polymers. When grown on agar medium, CMC degradation occurred only directly below colonies of R. leguminosarum, suggesting that the enzymes remain attached to the bacteria. Unexpectedly, when a PlyA-PlyB-secreting colony was grown in close proximity to mutants unable to produce or secrete PlyA and PlyB, CMC degradation occurred below that part of the mutant colonies closest to the wild type. There was no CMC degradation in the region between the colonies. By growing PlyB-secreting colonies on a lawn of CMC-nondegrading mutants, we could observe a halo of CMC degradation around the colony. Using various mutant strains, we demonstrate that PlyB diffuses beyond the edge of the colony but does not degrade CMC unless it is in contact with the appropriate colony surface. PlyA appears to remain attached to the cells since no such diffusion of PlyA activity was observed. EPS defective mutants could secrete both PlyA and PlyB, but these enzymes were inactive unless they came into contact with an EPS(+) strain, indicating that EPS is required for activation of PlyA and PlyB. However, we were unable to activate CMC degradation with a crude EPS fraction, indicating that activation of CMC degradation may require an intermediate in EPS biosynthesis. Transfer of PlyB to Agrobacterium tumefaciens enabled it to degrade CMC, but this was only observed if it was grown on a lawn of R. leguminosarum. This indicates that the surface of A. tumefaciens is inappropriate to activate CMC degradation by PlyB. Analysis of CMC degradation by other rhizobia suggests that activation of secreted glycanases by surface components may occur in other species. PMID- 10671452 TI - Partitioning of the linear chromosome during sporulation of Streptomyces coelicolor A3(2) involves an oriC-linked parAB locus. AB - Candidate partitioning genes (parA and parB) for the linear chromosome of Streptomyces coelicolor were identified by DNA sequencing in a series of seven genes located between rnpA and trxA near the chromosomal replication origin. The most likely translation start point of parB overlapped the parA stop codon, suggestive of coregulation, and transcription analysis suggested that the two genes formed an operon. Deletion of part of parB had no effect on the growth or appearance of colonies but caused a deficiency in DNA partitioning during the multiple septation events involved in converting aerial hyphae into long chains of spores. At least 13% of spore compartments failed to inherit the normal DNA allocation. The same phenotype was obtained with a deletion removing a segment of DNA from both parA and parB. Reinforcing the idea of a special role for the par locus during sporulation, the stronger of two parAB promoters was greatly upregulated at about the time when sporulation septation was maximal in colonies. Three copies of a 14-bp inverted repeat (GTTTCACGTGAAAC) were found in or near the parAB genes, and at least 12 more identical copies were identified within 100 kb of oriC from the growing genome sequence database. Only one perfect copy of the 14-bp sequence was present in approximately 5 Mb of sequence available from the rest of the genome. The 14-bp sequence was similar to sequences identified as binding sites for Spo0J, a ParB homologue from Bacillus subtilis believed to be important for DNA partitioning (D. C.-H. Lin and A. D. Grossman, Cell 92:675-685, 1998). One of these sites encompassed the transcription start point of the stronger parA promoter. PMID- 10671453 TI - Novel phospholipase A activity secreted by Legionella species. AB - Bacterial phospholipases are regarded as a major virulence factor in infection. In bacteria associated with pneumonia, destruction of lung surfactant and host cell membranes by bacterial phospholipases secreted during infection is thought to contribute to the disease. Phospholipase C (PLC) activity has been described in several Legionella species (W. B. Baine, J. Gen. Microbiol. 134:489-498, 1988; W. B. Baine, J. Gen. Microbiol. 131:1383-1391, 1985). By using detection methods such as thin-layer chromatography and mass spectrometry, PLC activity could not be detected in several strains of Legionella pneumophila. Instead, phospholipid degradation was identified to be caused by a novel PLA activity. We could demonstrate that PLA secretion starts at the mid-exponential-growth phase when bacteria were grown in liquid culture. Several Legionella species secreted different amounts of PLA. Legionella PLA may act as a powerful agent in the mediation of pathogenicity due to destruction of lung surfactant and epithelial cells. PMID- 10671454 TI - Effect of temperature on stability and activity of elongation factor 2 proteins from Antarctic and thermophilic methanogens. AB - Despite the presence and abundance of archaea in low-temperature environments, little information is available regarding their physiological and biochemical properties. In order to investigate the adaptation of archaeal proteins to low temperatures, we purified and characterized the elongation factor 2 (EF-2) protein from the Antarctic methanogen Methanococcoides burtonii, which was expressed in Escherichia coli, and compared it to the recombinant EF-2 protein from a phylogenetically related thermophile, Methanosarcina thermophila. Using differential scanning calorimetry to assess protein stability and enzyme assays for the intrinsic GTPase activity, we identified biochemical and biophysical properties that are characteristic of the cold-adapted protein. This includes a higher activity at low temperatures caused by a decrease of the activation energy necessary for GTP hydrolysis and a decreased activation energy for the irreversible denaturation of the protein, which indicates a less thermostable structure. Comparison of the in vitro properties of the proteins with the temperature-dependent characteristics of growth of the organisms indicates that additional cytoplasmic factors are likely to be important for the complete thermal adaptation of the proteins in vivo. This is the first study to address thermal adaptation of proteins from a free-living, cold-adapted archaeon, and our results indicate that the ability of the Antarctic methanogen to adapt to the cold is likely to involve protein structural changes. PMID- 10671455 TI - Succinyl coenzyme A synthetase of Pseudomonas aeruginosa with a broad specificity for nucleoside triphosphate (NTP) synthesis modulates specificity for NTP synthesis by the 12-kilodalton form of nucleoside diphosphate kinase. AB - Pseudomonas aeruginosa secretes copious amounts of an exopolysaccharide called alginate during infection in the lungs of cystic fibrosis patients. A mutation in the algR2 gene of mucoid P. aeruginosa is known to exhibit a nonmucoid (nonalginate-producing) phenotype and showed reduced activities of succinyl coenzyme A (CoA) synthetase (Scs) and nucleoside diphosphate kinase (Ndk), implying coregulation of Ndk and Scs in alginate synthesis. We have cloned and characterized the sucCD operon encoding the alpha and beta subunits of Scs from P. aeruginosa and have studied the role of Scs in generating GTP, an important precursor in alginate synthesis. We demonstrate that, in the presence of GDP, Scs synthesizes GTP using ATP as the phosphodonor and, in the presence of ADP, Scs synthesizes ATP using GTP as a phosphodonor. In the presence of inorganic orthophosphate, succinyl-CoA, and an equimolar amount of ADP and GDP, Scs synthesizes essentially an equimolar amount of ATP and GTP. Such a mechanism of GTP synthesis can be an alternate source for the synthesis of alginate as well as for the synthesis of other macromolecules requiring GTP such as RNA and protein. Scs from P. aeruginosa is also shown to exhibit a broad NDP kinase activity. In the presence of inorganic orthophosphate (P(i)), succinyl-CoA, and either GDP, ADP, UDP or CDP, it synthesizes GTP, ATP, UTP, or CTP. Scs was previously shown to copurify with Ndk, presumably as a complex. In mucoid cells of P. aeruginosa, Ndk is also known to exist in two forms, a 16-kDa cytoplasmic form predominant in the log phase and a 12-kDa membrane-associated form predominant in the stationary phase. We have observed that the 16-kDa Ndk-Scs complex present in nonmucoid cells, synthesizes all three of the nucleoside triphosphates from a mixture of GDP, UDP, and CDP, whereas the 12-kDa Ndk-Scs complex specifically present in mucoid cell predominantly synthesizes GTP and UTP but not CTP. Such regulation may promote GTP synthesis in the stationary phase when the bulk of alginate is synthesized by mucoid P. aeruginosa. PMID- 10671456 TI - Characterization of SotA and SotB, two Erwinia chrysanthemi proteins which modify isopropyl-beta-D-thiogalactopyranoside and lactose induction of the Escherichia coli lac promoter. AB - The expression, in Escherichia coli, of variants of the Erwinia chrysanthemi secretion genes outB and outS under the Ptac promoter is toxic to the cells. During attempts to clone E. chrysanthemi genes able to suppress this toxicity, I identified two genes, sotA and sotB, whose products are able to reduce the isopropyl-beta-D-thiogalactopyranoside (IPTG) induction of the E. coli lac promoter. SotA and SotB belong to two different families of the major facilitator superfamily. SotA is a member of the sugar efflux transporter family, while SotB belongs to the multidrug efflux family. The results presented here suggest that SotA and SotB are sugar efflux pumps. SotA reduces the intracellular concentration of IPTG, lactose, and arabinose. SotB reduces the concentration of IPTG, lactose, and melibiose. Expression of sotA and sotB is not regulated by their substrates, but sotA is activated by the cyclic AMP receptor protein (CRP), while sotB is repressed by CRP. Lactose is weakly toxic for E. chrysanthemi. This toxicity is increased in a sotB mutant which cannot efficiently efflux lactose. This first evidence for a physiological role of sugar efflux proteins suggests that their function could be to reduce the intracellular concentration of toxic sugars or sugar metabolites. PMID- 10671457 TI - Feruloyl esterase activity of the Clostridium thermocellum cellulosome can be attributed to previously unknown domains of XynY and XynZ. AB - The cellulosome of Clostridium thermocellum is a multiprotein complex with endo- and exocellulase, xylanase, beta-glucanase, and acetyl xylan esterase activities. XynY and XynZ, components of the cellulosome, are composed of several domains including xylanase domains and domains of unknown function (UDs). Database searches revealed that the C- and N-terminal UDs of XynY and XynZ, respectively, have sequence homology with the sequence of a feruloyl esterase of strain PC-2 of the anaerobic fungus Orpinomyces. Purified cellulosomes from C. thermocellum were found to hydrolyze FAXX (O-(5-O-[(E)-feruloyl]-alpha-L-arabinofuranosyl)-(1-->3) O-beta-D- xyl opyranosyl-(1-->4)-D-xylopyranose) and FAX(3) (5-O-[(E)-feruloyl] [O-beta-D-xylopyranosyl-(1-->2)]-O-alpha-L- arabinofuranosyl-[1-->3])-O-beta-D xylopyranosyl-(1-->4)-D-xylopyranose) , yielding ferulic acid as a product, indicating that they have feruloyl esterase activity. Nucleotide sequences corresponding to the UDs of XynY and XynZ were cloned into Escherichia coli, and the expressed proteins hydrolyzed FAXX and FAX(3). The recombinant feruloyl esterase domain of XynZ alone (FAE(XynZ)) and with the adjacent cellulose binding domain (FAE-CBD(XynZ)) were characterized. FAE-CBD(XynZ) had a molecular mass of 45 kDa that corresponded to the expected product of the 1,203-bp gene. K(m) and V(max) values for FAX(3) were 5 mM and 12.5 U/mg, respectively, at pH 6.0 and 60 degrees C. PAX(3), a substrate similar to FAX(3) but with a p-coumaroyl group instead of a feruloyl moiety was hydrolyzed at a rate 10 times slower. The recombinant enzyme was active between pH 3 to 10 with an optimum between pH 4 to 7 and at temperatures up to 70 degrees C. Treatment of Coastal Bermuda grass with the enzyme released mainly ferulic acid and a lower amount of p-coumaric acid. FAE(XynZ) had similar properties. Removal of the 40 C-terminal amino acids, residues 247 to 286, of FAE(XynZ) resulted in protein without activity. Feruloyl esterases are believed to aid in a release of lignin from hemicellulose and may be involved in lignin solubilization. The presence of feruloyl esterase in the C. thermocellum cellulosome together with its other hydrolytic activities demonstrates a powerful enzymatic potential of this organelle in plant cell wall decomposition. PMID- 10671458 TI - Respiration of 2,4,6-trinitrotoluene by Pseudomonas sp. strain JLR11. AB - Under anoxic conditions Pseudomonas sp. strain JLR11 can use 2,4, 6 trinitrotoluene (TNT) as the sole N source, releasing nitrite from the aromatic ring and subsequently reducing it to ammonium and incorporating it into C skeletons. This study shows that TNT can also be used as a terminal electron acceptor in respiratory chains under anoxic conditions by Pseudomonas sp. strain JLR11. TNT-dependent proton translocation coupled to the reduction of TNT to aminonitrotoluenes has been observed in TNT-grown cells. This extrusion did not occur in nitrate-grown cells or in anaerobic TNT-grown cells treated with cyanide, a respiratory chain inhibitor. We have shown that in a membrane fraction prepared from Pseudomonas sp. strain JLR11 grown on TNT under anaerobic conditions, the synthesis of ATP was coupled to the oxidation of molecular hydrogen and to the reduction of TNT. This phosphorylation was uncoupled by gramicidin. Respiration by Pseudomonas sp. strain JLR11 is potentially useful for the biotreatment of TNT in polluted waters and soils, particularly in phytorhizoremediation, in which bacterial cells are transported to the deepest root zones, which are poor in oxygen. PMID- 10671459 TI - Gene families encoding phase- and size-variable surface lipoproteins of Mycoplasma hyorhinis. AB - A prototype family of seven genes encoding the variable surface lipoproteins (Vlps) of Mycoplasma hyorhinis is characterized in the pathogenic SK76 strain, using long-range PCR to amplify and analyze the single chromosomal region containing expressed genes vlpA to -G, each of which is subject to phase and size variation. Smaller families of vlp genes in subclones of SK76 or in another strain of M. hyorhinis, GDL, can be attributed to deletions of specific vlp genes from the prototype array described here. Two genes, vlpA and the newly revealed vlpG, contain repeat motifs in their 3' coding regions that differ from the short tandem repeats in other vlp genes yet retain structural features common to all vlp gene products. SK76 and GDL vlp gene families are similarly organized and show sequence similarity between corresponding individual vlp genes. In light of the extensive potential for diversity within the vlp gene system, such conservation provides a provisional basis to hypothesize that vlp genes may exist in specific arrays that endow selected functions while retaining common structural features required during phase-variable expression of this set of gene products. PMID- 10671460 TI - The UspA1 protein and a second type of UspA2 protein mediate adherence of Moraxella catarrhalis to human epithelial cells in vitro. AB - The UspA1 and UspA2 proteins of Moraxella catarrhalis are structurally related, are exposed on the bacterial cell surface, and migrate as very high-molecular weight complexes in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Previous analysis of uspA1 and uspA2 mutants of M. catarrhalis strain 035E indicated that UspA1 was involved in adherence of this organism to Chang conjunctival epithelial cells in vitro and that expression of UspA2 was essential for resistance of this strain to killing by normal human serum (C. Aebi, E. R. Lafontaine, L. D. Cope, J. L. Latimer, S. R. Lumbley, G. H. McCracken, Jr., and E. J. Hansen, Infect. Immun. 66:3113-3119, 1998). In the present study, isogenic uspA1, uspA2, and uspA1 uspA2 mutations were constructed in three additional M. catarrhalis strains: 012E, TTA37, and 046E. The uspA1 mutant of strain 012E had a decreased ability to attach to Chang cells. However, inactivation of the uspA1 gene in both strain TTA37 and strain 046E did not cause a significant decrease in attachment ability. Inactivation of the uspA2 gene of strain TTA37 did result in a loss of attachment ability. Nucleotide sequence analysis revealed that the predicted protein encoded by the uspA2 genes of both strains TTA37 and 046E had a N-terminal half that resembled the N-terminal half of UspA1 proteins, whereas the C-terminal half of this protein was nearly identical to those of previously characterized UspA2 proteins. The gene encoding this "hybrid" protein was designated uspA2H. PCR-based analysis revealed that approximately 20% of M. catarrhalis strains apparently possess a uspA2H gene instead of a uspA2 gene. The M. catarrhalis uspA1, uspA2, and uspA2H genes were cloned and expressed in Haemophilus influenzae cells, which were used to prove that both the UspA1 and UspA2H proteins can function as adhesins in vitro. PMID- 10671461 TI - Streptococcus gordonii biofilm formation: identification of genes that code for biofilm phenotypes. AB - Viridans streptococci, which include Streptococcus gordonii, are pioneer oral bacteria that initiate dental plaque formation. Sessile bacteria in a biofilm exhibit a mode of growth that is distinct from that of planktonic bacteria. Biofilm formation of S. gordonii Challis was characterized using an in vitro biofilm formation assay on polystyrene surfaces. The same assay was used as a nonbiased method to screen isogenic mutants generated by Tn916 transposon mutagenesis for defective biofilm formation. Biofilms formed optimally when bacteria were grown in a minimal medium under anaerobic conditions. Biofilm formation was affected by changes in pH, osmolarity, and carbohydrate content of the growth media. Eighteen biofilm-defective mutants of S. gordonii Challis were identified based on Southern hybridization with a Tn916-based probe and DNA sequences of the Tn916-flanking regions. Molecular analyses of these mutants showed that some of the genes required for biofilm formation are involved in signal transduction, peptidoglycan biosynthesis, and adhesion. These characteristics are associated with quorum sensing, osmoadaptation, and adhesion functions in oral streptococci. Only nine of the biofilm-defective mutants had defects in genes of known function, suggesting that novel aspects of bacterial physiology may play a part in biofilm formation. Further identification and characterization of biofilm-associated genes will provide insight into the molecular mechanisms of biofilm formation of oral streptococci. PMID- 10671462 TI - Substrate range and genetic analysis of Acinetobacter vanillate demethylase. AB - An Acinetobacter sp. genetic screen was used to probe structure-function relationships in vanillate demethylase, a two-component monooxygenase. Mutants with null, leaky, and heat-sensitive phenotypes were isolated. Missense mutations tended to be clustered in specific regions, most of which make known contributions to catalytic activity. The vanillate analogs m-anisate, m-toluate, and 4-hydroxy-3,5-dimethylbenzoate are substrates of the enzyme and weakly inhibit the metabolism of vanillate by wild-type Acinetobacter bacteria. PCR mutagenesis of vanAB, followed by selection for strains unable to metabolize vanillate, yielded mutant organisms in which vanillate metabolism is more strongly inhibited by the vanillate analogs. Thus, the procedure opens for investigation amino acid residues that may contribute to the binding of either vanillate or its chemical analogs to wild-type and mutant vanillate demethylases. Selection of phenotypic revertants following PCR mutagenesis gave an indication of the extent to which amino acid substitutions can be tolerated at specified positions. In some cases, only true reversion to the original amino acid was observed. In other examples, a range of amino acid substitutions was tolerated. In one instance, phenotypic reversion failed to produce a protein with the original wild-type sequence. In this example, constraints favoring certain nucleotide substitutions appear to be imposed at the DNA level. PMID- 10671463 TI - Regulation of the cnr cobalt and nickel resistance determinant from Ralstonia sp. strain CH34. AB - Ralstonia sp. strain CH34 is resistant to nickel and cobalt cations. Resistance is mediated by the cnr determinant located on plasmid pMOL28. The cnr genes are organized in two clusters, cnrYXH and cnrCBA. As revealed by reverse transcriptase PCR and primer extension, transcription from these operons is initiated from promoters located upstream of the cnrY and cnrC genes. These two promoters exhibit conserved sequences at the -10 (CCGTATA) and -35 (CRAGGGGRAG) regions. The CnrH gene product, which is required for expression of both operons, is a sigma factor belonging to the sigma L family, whose activity seems to be governed by the membrane-bound CnrY and CnrX gene products in response to Ni(2+). Half-maximal activation from the cnrCBA operon was determined by using appropriate lacZ gene fusions and was shown to occur at an Ni(2+) concentration of about 50 microM. PMID- 10671464 TI - Regulation of the cnr cobalt and nickel resistance determinant of Ralstonia eutropha (Alcaligenes eutrophus) CH34. AB - The linked resistance to nickel and cobalt of Ralstonia eutropha-like strain CH34 (Alcaligenes eutrophus CH34) is encoded by the cnr operon, which is localized on the megaplasmid pMOL28. The regulatory genes cnrYXH have been cloned, overexpressed, and purified in Escherichia coli. CnrY fractionated as a 10.7-kDa protein in in vitro translation assays. CnrX, a periplasmic protein of 16.5 kDa, was overproduced and purified as a histidine-tagged fusion protein in E. coli. His-CnrX was found to possess a secondary structure content rich in alpha-helical and beta-sheet structures. CnrH, a sigma factor of the extracytoplasmic function family, was purified as an N-terminally histidine-tagged fusion. In gel shift mobility assays, His-CnrH, in the presence of E. coli core RNA polymerase enzyme, could retard at least two different promoter DNA targets, cnrYp and cnrHp, localized within the cnrYXH locus. These promoters and their transcription start sites were confirmed by primer extension. Purified His-CnrX did not inhibit the DNA-binding activity of His-CnrH and is therefore unlikely to be an anti-sigma factor, as previously hypothesized (EMBL M91650 description entry). To study the transcriptional response of the regulatory locus to metals and to probe promoter regions, transcriptional fusions were constructed between fragments of cnrYXH and the luxCDABE, luciferase reporter genes. Nickel and cobalt specifically induced the cnrYXH-luxCDABE fusion at optimal concentrations of 0.3 mM Ni(2+) and 2.0 mM Co(2+) in a noncomplexing medium for metals. The two promoter regions P(Y) (upstream cnrY) and P(H) (upstream cnrH) were probed and characterized using this vector and were found to control the nickel-inducible regulatory response of the cnr operon. The cnrHp promoter was responsible for full transcription of the cnrCBA structural resistance genes, while the cnrYp promoter was necessary to obtain metal-inducible transcription from the cnrHp promoter. The zinc resistance phenotype (ZinB) of a spontaneous cnr mutant strain, AE963, was investigated and could be attributed to an insertion of IS1087, a member of the IS2 family of insertion elements, within the cnrY gene. PMID- 10671465 TI - Influence of mutations in the mexR repressor gene on expression of the MexA-MexB oprM multidrug efflux system of Pseudomonas aeruginosa. AB - Several nalB-type multidrug-resistant mutants of Pseudomonas aeruginosa overexpressed MexAB-OprM and carried mutations in the local regulatory gene, mexR. Others, dubbed nalC types, carried mutations elsewhere and overexpressed MexAB-OprM less extensively than the nalB strains. Available evidence showed that MexR acted solely as repressor. Disruption of the mexR gene at various places suggested that the 5' end of mexR may be a part of the mexAB-oprM promoter. PMID- 10671466 TI - Physical map of the chromosome of the apple proliferation phytoplasma. AB - A physical map of the apple proliferation phytoplasma strain AT chromosome was constructed from genomic DNA extracted from diseased tobacco plants. The map was generated with single and double digestions of the chromosome with BssHII, SmaI, MluI, and ApaI restriction endonucleases and resolving the fragments by pulsed field gel electrophoresis. Partial digestion and Southern blot analysis were used to assist in the arrangement of the 14 contiguous restriction fragments obtained. From the restriction fragments generated by double digestions, the size of the circular chromosome was calculated to be approximately 645 kb. Locations of the two rRNA operons, the operon including the fus and tuf genes, and three other genes were placed on the map. Genome sizes and BssHII restriction profiles of apple proliferation strain AP15 and the pear decline and European stone fruit yellows phytoplasmas were different from that of strain AT. PMID- 10671467 TI - Effects of calcium and calcium chelators on growth and morphology of Escherichia coli L-form NC-7. AB - Growth of a wall-less, L-form of Escherichia coli specifically requires calcium, and in its absence, cells ceased dividing, became spherical, swelled, developed large vacuoles, and eventually lysed. The key cell division protein, FtsZ, was present in the L-form at a concentration five times less than that in the parental strain. One interpretation of these results is that the L-form possesses an enzoskeleton partly regulated by calcium. PMID- 10671468 TI - Cpx two-component signal transduction in Escherichia coli: excessive CpxR-P levels underlie CpxA* phenotypes. AB - In Escherichia coli, the CpxA-CpxR two-component signal transduction system and the sigma(E) and sigma(32) response pathways jointly regulate gene expression in adaptation to adverse conditions. These include envelope protein distress, heat shock, oxidative stress, high pH, and entry into stationary phase. Certain mutant versions of the CpxA sensor protein (CpxA* proteins) exhibit an elevated ratio of kinase to phosphatase activity on CpxR, the cognate response regulator. As a result, CpxA* strains display numerous phenotypes, many of which cannot be easily related to currently known functions of the CpxA-CpxR pathway. It is unclear whether CpxA* phenotypes are caused solely by hyperphosphorylation of CpxR. We here report that all of the tested CpxA* phenotypes depend on elevated levels of CpxR-P and not on cross-signalling of CpxA* to noncognate response regulators. PMID- 10671469 TI - Requirement for homologous recombination functions for expression of the mutA mistranslator tRNA-induced mutator phenotype in Escherichia coli. AB - Expression of the Escherichia coli mutA mutator phenotype requires recA, recB, recC, ruvA, and ruvC gene, but not recD, recF, recO, or recR genes. Thus, the recBCD-dependent homologous recombination system is a component of the signal pathway that activates an error-prone DNA polymerase in mutA cells. PMID- 10671470 TI - Novel missense mutations that affect the transport function of MalK, the ATP binding-cassette subunit of the Salmonella enterica serovar typhimurium maltose transport system. AB - We report on novel mutations in the malK gene of Salmonella enterica serovar Typhimurium, encoding the ATPase subunit of the maltose transporter (MalFGK(2)). Biochemical analysis suggests that (i) L86 might be involved in a signaling step during substrate translocation and (ii) E306 may be critical for the structural integrity of the protein. PMID- 10671471 TI - Mutational analysis of ligand recognition by tcp, the citrate chemoreceptor of Salmonella enterica serovar typhimurium. AB - The chemoreceptor Tcp mediates taxis to citrate. To identify citrate-binding residues, we substituted cysteine for seven basic or polar residues that are chosen based on the comparison of Tcp with the well-characterized chemoreceptors. The results suggest that Arg-63, Arg-68, Arg-72, Lys-75, and Tyr-150 (and probably other unidentified residues) are involved in the recognition of citrate. PMID- 10671472 TI - Cytochrome c' from Rhodobacter capsulatus confers increased resistance to nitric oxide. AB - We report the cloning and sequencing of the gene containing cytochrome c' (cycP) from the photosynthetic purple bacterium Rhodobacter capsulatus and the regions flanking that gene. Mutant strains unable to synthesize cytochrome c' had increased sensitivity to nitrosothiols and to nitric oxide (which binds to the heme moiety of cytochrome c'). PMID- 10671473 TI - Utilization of subsidiary chromosomal replication terminators in Bacillus subtilis. AB - The Bacillus subtilis merodiploid strain GSY1127 contains a large nontandem duplication of a portion of its chromosome within its left (anticlockwise) replication segment. This causes displacement of the replication terminus region to a noticeably asymmetric location relative to oriC. The utilization of the subsidiary replication terminators, TerIII and TerV, in the merodiploid strain has been compared with that in B. subtilis 168. It is shown that TerIII is utilized to a significant extent in GSY1127 and that TerV is used only marginally at the most. Neither of these terminators is used to a measurable extent in the 168 strain. It is concluded that TerIII and TerV do indeed function as backups to the major terminator TerI, as has been generally thought. It is further concluded that, in the 168 strain, the vast majority of clockwise forks are arrested at the highly efficient TerI terminator, with fork fusion between the approaching forks occurring frequently while the clockwise fork is stationary at TerI. PMID- 10671474 TI - Stress triggers a process that limits activation of the Bacillus subtilis stress transcription factor sigma(B). AB - Stress-induced activation of the Bacillus subtilis transcription factor sigma(B) is transitory. To determine whether the process that limits sigma(B) activation is itself triggered by stress, B. subtilis strains in which the stress pathway was artificially activated by the induced expression of a positive regulatory protein (RsbT) were exposed to ethanol stress and were monitored for the persistence of sigma(B) activity. Without ethanol treatment, the induced cultures displayed continuously high sigma(B) activity. Ethanol treatment restricted ongoing sigma(B) activity, but only in strains with intact rsbX and -S genes. The loss of other gene products (RsbR and Obg) known to participate in the stress activation pathway had little influence in blocking the ethanol effect. The data argue that stress upregulates the activity of the RsbX-S regulatory pair to restrict sigma(B) induction following stress. PMID- 10671475 TI - A tripartite nuclear localization signal in the PDZ-domain protein L-periaxin. AB - The murine Periaxin gene encodes two PDZ-domain proteins in myelin-forming Schwann cells of the vertebrate peripheral nervous system (Dytrych, L., Sherman, D. L., Gillespie, C. S., and Brophy, P. J. (1998) J. Biol. Chem. 273, 5794-5800). Here we show that L-periaxin is targeted to the nucleus of embryonic Schwann cells. Subsequently, the protein redistributes to the plasma membrane processes of the myelinating Schwann cell where it is believed to function in a signaling complex. In contrast, L-periaxin remains in the nucleus when expressed ectopically in oligodendrocytes, the myelin-forming glia of the central nervous system. The nuclear localization signal (NLS) is basic and tripartite and comprises three signals that act synergistically. Nuclear targeting of L-periaxin is energy-dependent and is inhibited by cell-cell contact. These data show that L periaxin is a member of a growing family of proteins that can shuttle between the nucleus and cortical signaling/adherence complexes. PMID- 10671476 TI - Activation of human T lymphocytes is inhibited by peroxisome proliferator activated receptor gamma (PPARgamma) agonists. PPARgamma co-association with transcription factor NFAT. AB - T lymphocyte activation is highlighted by the induction of interleukin-2 (IL-2) gene expression, which governs much of the early lymphocyte proliferation responses. Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the nuclear receptor superfamily of ligand-activated transcription factors. PPARgamma mRNA expression was found in human peripheral blood T lymphocytes, raising the possibility of PPARgamma involvement in the regulation of T cell function. Here we show that PPARgamma ligands, troglitazone and 15 deoxy-Delta(12,14) prostaglandin J(2), but not PPARalpha agonist Wy14643, inhibited IL-2 production and phytohemagglutinin-inducible proliferation in human peripheral blood T-cells in a dose-dependent manner. This inhibitory effect on IL 2 was restricted to the PPARgamma2-expressing, not the PPARgamma-lacking, subpopulation of transfected Jurkat cells. The activated PPARgamma physically associates with transcriptional factor NFAT regulating the IL-2 promoter, blocking NFAT DNA binding and transcriptional activity. This interaction with T cell-specific transcription factors indicates an important immunomodulatory role for PPARgamma in T lymphocytes and could suggest a previously unrecognized clinical potential for PPARgamma ligands as immunotherapeutic drugs to treat T cell-mediated diseases by targeting IL-2 gene expression. PMID- 10671477 TI - Stimulation of slow skeletal muscle fiber gene expression by calcineurin in vivo. AB - Adult skeletal muscle fibers can be categorized into fast and slow twitch subtypes based on specialized contractile and metabolic properties and on distinctive patterns of muscle gene expression. Muscle fiber-type characteristics are dependent on the frequency of motor nerve stimulation and are thought to be controlled by calcium-dependent signaling. The calcium, calmodulin-dependent protein phosphatase, calcineurin, stimulates slow fiber-specific gene promoters in cultured skeletal muscle cells, and the calcineurin inhibitor, cyclosporin A, inhibits slow fiber gene expression in vivo, suggesting a key role of calcineurin in activation of the slow muscle fiber phenotype. Calcineurin has also been shown to induce hypertrophy of cardiac muscle and to mediate the hypertrophic effects of insulin-like growth factor-1 on skeletal myocytes in vitro. To determine whether activated calcineurin was sufficient to induce slow fiber gene expression and hypertrophy in adult skeletal muscle in vivo, we created transgenic mice that expressed activated calcineurin under control of the muscle creatine kinase enhancer. These mice exhibited an increase in slow muscle fibers, but no evidence for skeletal muscle hypertrophy. These results demonstrate that calcineurin activation is sufficient to induce the slow fiber gene regulatory program in vivo and suggest that additional signals are required for skeletal muscle hypertrophy. PMID- 10671478 TI - Isolation, structural characterization, and bioactivity of a novel neuromedin U analog from the defensive skin secretion of the Australasian tree frog, Litoria caerulea. AB - We report the isolation of a novel bioactive peptide, neuromedin U-23 (NmU-23), from the defensive skin secretion of the Australasian tree frog, Litoria caerulea. The primary structure of the peptide was established by a combination of microsequencing, mass spectroscopy and site-directed antiserum immunoreactivity as SDEEVQVPGGVISNGYFLFRPRN-amide (M(r) 2580.6). A synthetic replicate of frog NmU-23 displaced monoradioiodinated rat NmU-23 from uterine membranes in a dose-dependent fashion indistinguishable from nonisotopically labeled rat NmU-23. In a rat uterine smooth muscle strip preparation, synthetic frog NmU-23 produced dose-dependent contractions identical to porcine NmU-25. However, in a preparation of human urinary bladder muscle strip, the synthetic frog peptide was more potent than porcine NmU-25 in eliciting contraction and produced desensitization of the preparation to the latter peptide. This report demonstrates that the defensive skin secretion of a frog contains a novel peptide exhibiting a high degree of primary structural similarity to the endogenous vertebrate peptide, NmU, and that this frog skin analog displays biological activity in mammalian tissues. PMID- 10671479 TI - Fhit-nucleotide specificity probed with novel fluorescent and fluorogenic substrates. AB - Fhit, a member of the histidine triad superfamily of nucleotide-binding proteins, binds and cleaves diadenosine polyphosphates and functions as a tumor suppressor in human epithelial cancers. Function of Fhit in tumor suppression does not require diadenosine polyphosphate cleavage but correlates with the ability to form substrate complexes. As diadenosine polyphosphates are at lower cellular concentrations than mononucleotides, we sought to quantify interactions between Fhit and competitive inhibitors with the use of diadenosine polyphosphate analogs containing fluorophores in place of one nucleoside. Appp-S-(7-diethylamino-4 methyl-3-(4-succinimidylphenyl)) coumarin (ApppAMC), Appp-S-(4-4-difluoro-5,7 dimethyl-4-bora-3a, 4a-diaza-s-indacine-3-yl) methylaminoacetyl (ApppBODIPY), and GpppBODIPY, synthesized in high yield, are effective Fhit substrates, producing AMP or GMP plus fluorophore diphosphates. GpppBODIPY cleavage is accompanied by a 5.4-fold increase in fluorescence because BODIPY fluorescence is quenched by stacking with guanine. Titration of unlabeled diadenosine polyphosphates, inorganic pyrophosphate, mononucleotides, and inorganic phosphate into fluorescent assays provided values of K(m) and K(I) as competitive inhibitors. The data indicate that Fhit discriminates between good substrates via k(cat) and against cellular competitors in equilibrium binding terms. Surprisingly, pyrophosphate competes better than purine mononucleotides. PMID- 10671480 TI - Plasma membrane calcium channels in human carcinoma A431 cells are functionally coupled to inositol 1,4,5-trisphosphate receptor-phosphatidylinositol 4,5 bisphosphate complexes. AB - In most nonexcitable cells, calcium (Ca(2+)) release from inositol 1,4,5 trisphosphate (InsP(3))-sensitive intracellular Ca(2+) stores is coupled to Ca(2+) influx (calcium release-activated channels (I(CRAC))) pathway. Despite intense investigation, the molecular identity of I(CRAC) and the mechanism of its activation remain poorly understood. InsP(3)-dependent miniature calcium channels (I(min)) display functional properties characteristic for I(CRAC). Here we used patch clamp recordings of I(min) channels in human carcinoma A431 cells to demonstrate that I(min) activity was greatly enchanced in the presence of anti phosphatidylinositol 4, 5-bisphosphate antibody (PIP(2)Ab) and diminished in the presence of PIP(2). Anti-PIP(2) antibody induced a greater than 6-fold increase in I(min) sensitivity for InsP(3) activation and an almost 4-fold change in I(min) maximal open probability. The addition of exogenous PIP(2) vesicles to the cytosolic surface of inside-out patches inhibited I(min) activity. These results lead us to propose an existence of a Ca(2+) influx pathway in nonexcitable cells activated via direct conformational coupling with a selected population of InsP(3) receptors, located just underneath the plasma membrane and coupled to PIP(2). The described pathway provides for a highly compartmentalized Ca(2+) influx and intracellular Ca(2+) store refilling mechanism. PMID- 10671481 TI - Temperature-dependent chaperone activity and structural properties of human alphaA- and alphaB-crystallins. AB - The chaperone activity and biophysical properties of recombinant human alphaA- and alphaB-crystallins were studied by light scattering and spectroscopic methods. While the chaperone function of alphaA-crystallin markedly improves with an increase in temperature, the activity of alphaB homopolymer appears to change very little upon heating. Compared with alphaB-crystallin, the alphaA-homopolymer is markedly less active at low temperatures, but becomes a more active species at high temperatures. At physiologically relevant temperatures, the alphaB homopolymer appears to be modestly (two times or less) more potent chaperone than alphaA homopolymer. In contrast to very similar thermotropic changes in the secondary structure of both homopolymers, alphaA- and alphaB-crystallins markedly differ with respect to the temperature-dependent surface hydrophobicity profiles. Upon heating, alphaA-crystallin undergoes a conformational transition resulting in the exposure of additional hydrophobic sites, whereas no such transition occurs for alphaB-crystallin. The correlation between temperature-dependent changes in the chaperone activity and hydrophobicity properties of the individual homopolymers supports the view that the chaperone activity of alpha-crystallin is dependent on the presence of surface-exposed hydrophobic patches. However, the present data also show that the surface hydrophobicity is not the sole determinant of the chaperone function of alpha-crystallin. PMID- 10671483 TI - Studies on the interleukin-6-type cytokine signal transducer gp130 reveal a novel mechanism of receptor activation by monoclonal antibodies. AB - The transmembrane glycoprotein gp130 belongs to the family of hematopoietic cytokine receptors. It represents the common signal transducing receptor component of the so called interleukin-6-type cytokines. For several cytokine receptors including gp130 it has been shown that receptor activation cannot only be achieved by the natural ligand but also by single monoclonal antibodies raised against the receptor ectodomain. These findings have been interpreted in a way that dimerization of cytokine receptors is sufficient for receptor activation. Here we show that the recently described gp130-activating antibody B-S12 actually consists of two different monoclonal antibodies. By subcloning of B-S12 the monoclonal antibodies B-S12-A5 and B-S12-G7 were obtained. The individual antibodies are biologically inactive, in combination they exert B-S12-like activity on hepatoma cells. On Ba/F3 cells stably transfected with gp130 a combination of B-S12-G7 with another monoclonal gp130 antibody, B-P8, is required to stimulate proliferation. Using gp130 deletion mutants we show that all three antibodies map to domains 2 and 3 of gp130 which constitute the cytokine binding module. The individual antibodies inhibit activation of the signal transducer by interleukin-6 and interfere with binding of interleukin-6 to gp130. Interestingly, the combination of B-S12-G7 and a Fab fragment of B-P8 retains biological activity. We conclude from our data that (i) the monoclonal antibodies activate gp130 by mimicking the natural ligand and (ii) enforcement of gp130 dimerization is not sufficient for receptor activation but additional conformational requirements have to be fulfilled. PMID- 10671482 TI - Identification of Cox20p, a novel protein involved in the maturation and assembly of cytochrome oxidase subunit 2. AB - We have identified Cox20p, a 23.8-kDa protein of the mitochondrial inner membrane that is involved in the biogenesis of the yeast cytochrome oxidase complex. Cytochrome oxidase subunit 2 (Cox2p) accumulates as a precursor in cox20 mutants, suggesting a defect in biogenesis of this mitochondrially encoded protein. The inability of cox20 mutants to process the subunit 2 precursor (pCox2p) is not due to impaired export of the protein across the inner membrane or to an inactive Imp1p/Imp2p peptidase. Rather, Cox20p specifically binds the newly synthesized pCox2p, a step required to present the exported pCox2p as a substrate to the Imp1p peptidase. All of the endogenous pCox2p accumulated in an Deltaimp1 mutant, and a small fraction of Cox2p in wild type yeast, is detected in a complex with Cox20p. Following maturation Cox2p remained associated with Cox20p, prior to assembling into the cytochrome oxidase complex. We propose that Cox20p acts as a membrane-bound chaperone necessary for cleavage of pCox2p and for interaction of the mature protein with other subunits of cytochrome oxidase in a later step of the assembly process. PMID- 10671484 TI - Partial occlusion of both cavities of the eukaryotic chaperonin with antibody has no effect upon the rates of beta-actin or alpha-tubulin folding. AB - The eukaryotic chaperonin containing T-complex polypeptide 1 (CCT) is required in vivo for the production of native actin and tubulin. It is a 900-kDa oligomer formed from two back-to-back rings, each containing eight different subunits surrounding a central cavity in which interactions with substrates are thought to occur. Here, we show that a monoclonal antibody recognizing the C terminus of the CCTalpha subunit can bind inside, and partially occlude, both cavities of apo CCT. Rabbit reticulocyte lysate was programmed to synthesize beta-actin and alpha tubulin in the presence and absence of anti-CCTalpha antibody. The binding of the antibody inside the cavity and its occupancy of a large part of it does not prevent the folding of beta-actin and alpha-tubulin by CCT, despite the fact that all the CCT in the in vitro translation reactions was continuously bound by two antibody molecules. Furthermore, no differences in the protease susceptibility of actin bound to CCT in the presence and absence of the monoclonal antibody were detected, indicating that the antibody molecules do not perturb the conformation of actin folding intermediates substantially. These data indicate that complete sequestration of substrate by CCT may not be required for productive folding, suggesting that there are differences in its folding mechanism compared with the Group I chaperonins. PMID- 10671485 TI - Trypsin stimulates integrin alpha(5)beta(1)-dependent adhesion to fibronectin and proliferation of human gastric carcinoma cells through activation of proteinase activated receptor-2. AB - Trypsin is widely expressed in various non-pancreatic tissues at low levels and overexpressed in some types of human cancers. In the present study, we found that trypsin stimulates integrin-dependent adhesion and growth of MKN-1 human gastric carcinoma cells. MKN-1 cells expressed both proteinase-activated receptor-1 (PAR 1) and PAR-2, which are activated by thrombin and trypsin, respectively. Both trypsin and the PAR-2 ligand SLIGKV promoted integrin alpha(5)beta(1)-mediated adhesion of MKN-1 cells to fibronectin, and less effectively integrin alpha(v)beta(3)-mediated cell adhesion to vitronectin, but not that to type IV collagen or laminin-1 at all. Thrombin and the PAR-1 ligand SFLLRN promoted the cell adhesion to vitronectin more strongly than trypsin or the PAR-2 ligand, but not the cell adhesion to fibronectin at all. The cell adhesion-stimulating effect of the PAR-2 ligand was significantly reduced by the pre-treatment of cells with trypsin, indicating that the effect of trypsin is mediated by PAR-2 activation. The trypsin-stimulated cell adhesion to vitronectin, but not to fibronectin, was effectively inhibited by the G(i) protein blocker pertussis toxin, and both cell adhesions were completely inhibited by the Src kinase inhibitor herbimycin A. Furthermore, trypsin and the PAR-2 ligand stimulated growth of MKN-1 cells more strongly than thrombin or the PAR-1 ligand. These results show that trypsin regulates cellular adhesion and proliferation by inducing PAR-2/G protein signalings, and that the integrin alpha(5)beta(1)- and integrin alpha(v)beta(3) dependent cell adhesions are regulated by different PAR/G protein signalings. PMID- 10671486 TI - Elucidation of the structural features of heparan sulfate important for interaction with the Hep-2 domain of fibronectin. AB - The interaction of fibronectin with cell surface heparan sulfate proteoglycans is important biologically in inducing reorganization of the cytoskeleton and the assembly of focal adhesions. The major heparan sulfate-binding site in fibronectin, which is also implicated in these morphological events, is the COOH terminal Hep-2 domain. We describe the first extensive study of the structural determinants required for the interaction between heparan sulfate/heparin and Hep 2. It is clear that, in heparan sulfate, there is a very prominent role for N sulfate groups, as opposed to a relatively small apparent contribution from carboxyl groups. Furthermore, a minimal octasaccharide binding sequence appeared to contain at least two 2-O-sulfated iduronate residues, but no 6-O-sulfate groups. However, affinity was enhanced by the presence of 6-O-sulfates, and the interaction with Hep-2 also increased progressively with oligosaccharide size up to a maximum length of a tetradecasaccharide. This overall specificity is compatible with recent information on the structure of Hep-2 (Sharma, A., Askari, J. A., Humphries, M. J., Jones, E. Y., and Stuart, D. I. (1999) EMBO J. 18, 1468 1479) in which two separate, positively charged clusters, involving up to 11 basic amino acid residues (mostly arginines with their preferential ability to co ordinate sulfate groups), could form a single extended binding site. PMID- 10671487 TI - GLUTX1, a novel mammalian glucose transporter expressed in the central nervous system and insulin-sensitive tissues. AB - Based on homology with GLUT1-5, we have isolated a cDNA for a novel glucose transporter, GLUTX1. This cDNA encodes a protein of 478 amino acids that shows between 29 and 32% identity with rat GLUT1-5 and 32-36% identity with plant and bacterial hexose transporters. Unlike GLUT1-5, GLUTX1 has a short extracellular loop between transmembrane domain (TM) 1 and TM2 and a long extracellular loop between TM9 and TM10 that contains the only N-glycosylation site. When expressed in Xenopus oocytes, GLUTX1 showed strong transport activity only after suppression of a dileucine internalization motif present in the amino-terminal region. Transport activity was inhibited by cytochalasin B and partly competed by D-fructose and D-galactose. The Michaelis-Menten constant for glucose was approximately 2 mM. When translated in reticulocytes lysates, GLUTX1 migrates as a 35-kDa protein that becomes glycosylated in the presence of microsomal membranes. Western blot analysis of GLUTX1 transiently expressed in HEK293T cells revealed a diffuse band with a molecular mass of 37-50 kDa that could be converted to a approximately 35-kDa polypeptide following enzymatic deglycosylation. Immunofluorescence microscopy detection of GLUTX1 transfected into HEK293T cells showed an intracellular staining. Mutation of the dileucine internalization motif induced expression of GLUTX1 at the cell surface. GLUTX1 mRNA was detected in testis, hypothalamus, cerebellum, brainstem, hippocampus, and adrenal gland. We hypothesize that, in a similar fashion to GLUT4, in vivo cell surface expression of GLUTX1 may be inducible by a hormonal or other stimulus. PMID- 10671488 TI - The ubiquitin-related BAG-1 provides a link between the molecular chaperones Hsc70/Hsp70 and the proteasome. AB - The BAG-1 protein modulates the chaperone activity of Hsc70 and Hsp70 in the mammalian cytosol and nucleus. Remarkably, BAG-1 possesses a ubiquitin-like domain at its amino terminus, suggesting a link to the ubiquitin/proteasome system. Here we show that BAG-1 is indeed associated with the 26 S proteasome in HeLa cells. Binding of the chaperone cofactor to the proteolytic complex is regulated by ATP hydrolysis and is not mediated by Hsc70 and Hsp70. The presented findings reveal a role of BAG-1 as a physical link between the Hsc70/Hsp70 chaperone system and the proteasome. In fact, targeting of BAG-1 to the proteasome promotes an association of the chaperones with the proteolytic complex in vitro and in vivo. A regulatory function of the chaperone cofactor at the interface between protein folding and protein degradation is thus indicated. PMID- 10671489 TI - A redox mechanism controls differential DNA binding activities of hypoxia inducible factor (HIF) 1alpha and the HIF-like factor. AB - Hypoxia-inducible factor 1alpha (HIF-1alpha) and the HIF-like factor (HLF) are two highly related basic Helix-Loop-Helix/Per-Arnt-Sim (bHLH/PAS) homology transcription factors that undergo dramatically increased function at low oxygen levels. Despite strong similarities in their activation mechanisms (e.g. they both undergo rapid hypoxia-induced protein stabilization, bind identical target DNA sequences, and induce synthetic reporter genes to similar degrees), they are both essential for embryo survival via distinct functions during vascularization (HIF-1alpha) or catecholamine production (HLF). It is currently unknown how such specificity of action is achieved. We report here that DNA binding by HLF, but not by HIF-1alpha, is dependent upon reducing redox conditions. In vitro DNA binding and mammalian two-hybrid assays showed that a unique cysteine in the DNA binding basic region of HLF is a target for the reducing activity of redox factor Ref-1. Although the N-terminal DNA-binding domain of HIF-1alpha can function in the absence of Ref-1, we found that the C-terminal region containing the transactivation domain requires Ref-1 for full activity. Our data reveal that the hypoxia-inducible factors are subject to complex redox control mechanisms that can target discrete regions of the proteins and are the first to establish a discriminating control mechanism for differential regulation of HIF-1alpha and HLF activity. PMID- 10671490 TI - Function of the membrane fusion protein, MexA, of the MexA, B-OprM efflux pump in Pseudomonas aeruginosa without an anchoring membrane. AB - Resistance of Pseudomonas aeruginosa to multiple species of antibiotics is largely attributable to expression of the MexA, B-OprM efflux pump. The MexA protein is thought to be located at the inner membrane and has been assumed to link the xenobiotics-exporting subunit, MexB, and the outer membrane channel protein, OprM. To verify this assumption, we analyzed membrane anchoring and localization of the MexA protein. n-[9, 10-(3)H]Palmitic acid incorporation experiments revealed that MexA was radiolabeled with palmitic acid, suggesting that the MexA anchors the inner membrane via the fatty acid moiety. To evaluate the role of lipid modification and inner membrane anchoring, we substituted cysteine 24 with phenylalanine or tyrosine and tested whether or not these mutant MexAs function properly. When the mutant mexAs were expressed in the strain lacking chromosomal mexA in the presence of n-[9,10-(3)H]palmitic acid, we found undetectable radiolabeling at the MexA band. These transformants restored antibiotic resistance to the level of the wild-type strain, indicating that lipid modification is not essential for MexA function. These mutant strains contained both processed and unprocessed forms of the MexA proteins. Cellular fractionation experiments revealed that an unprocessed form of MexA anchored the inner membrane probably via an uncleaved signal sequence, whereas the processed form was undetectable in the membrane fraction. To assure that the lipid-free MexA polypeptide could be unbound to the membrane, we analyzed the two-dimensional membrane topology by the gene fusion technique. A total of 78 mexA-blaM fusions covering the entire MexA polypeptide were constructed, and all fusion sites were shown to be located at the periplasm. To answer the question of whether or not membrane anchoring is essential for the MexA function, we replaced the signal sequence of the MexA protein with that of the azurin protein, which contains a cleavable signal sequence but no lipid modification site. The signal sequence of the azurin-MexA hybrid protein was properly processed and bore the mature MexA, which was fully recovered in the soluble fraction. The transformant, which expressed azurin-MexA hybrid protein restored the antibiotic resistance to a level indistinguishable from that of the wild-type strain. We concluded from these results that the MexA protein is fully functional as expressed in the periplasmic space without anchoring the inner membrane. This finding questioned the assumption that the membrane fusion proteins connect the inner and outer membranes. PMID- 10671491 TI - N(alpha)-acetylation and proteolytic activity of the yeast 20 S proteasome. AB - N(alpha)-acetylation, catalyzed co-translationally with N(alpha) acetyltransferase (NAT), is the most common modifications of eukaryotic proteins. In yeast, there are at least three NATs: NAT1, MAK3, and NAT3. The 20 S proteasome subunits were purified from the normal strain and each of the deletion mutants, nat1, mak3, and nat3. The electrophoretic mobility of these subunits was compared by two-dimensional gel electrophoresis. Shifts toward the alkaline side of the gel and unblocking of the N terminus of certain of the subunits in one or another of the mutants indicated that the alpha1, alpha2, alpha3, alpha4, alpha7, and beta3 subunits were acetylated with NAT1, the alpha5 and alpha6 subunits were acetylated with MAK3, and the beta4 subunit was acetylated with NAT3. Furthermore, the Ac-Met-Phe-Leu and Ac-Met-Phe-Arg termini of the alpha5 and alpha6 subunits, respectively, extended the known types of MAK3 substrates. Thus, nine subunits were N (alpha)-acetylated, whereas the remaining five were processed, resulting in the loss of the N-terminal region. The 20 S proteasomes derived from either the nat1 mutant or the normal strain were similar in respect to chymotrypsin-like, trypsin-like, and peptidylglutamyl peptide hydrolyzing activities in vitro, suggesting that N(alpha)-acetylation does not play a major functional role in these activities. However, the chymotrypsin-like activity in the absence of sodium dodecyl sulfate was slightly higher in the nat1 mutant than in the normal strain. PMID- 10671492 TI - The low M(r) protein-tyrosine phosphatase is involved in Rho-mediated cytoskeleton rearrangement after integrin and platelet-derived growth factor stimulation. AB - The low molecular weight protein-tyrosine phosphatase (LMW-PTP) is an enzyme that is involved in the early events of platelet-derived growth factor (PDGF) receptor signal transduction. In fact, LMW-PTP is able to specifically bind and dephosphorylate activated PDGF receptor, thus modulating PDGF-induced mitogenesis. In particular, LMW-PTP is involved in pathways that regulate the transcription of the immediately early genes myc and fos in response to growth factor stimulation. Recently, we have found that LMW-PTP exists constitutively in cytosolic and cytoskeleton-associated localization and that, after PDGF stimulation, c-Src is able to bind and phosphorylate LMW-PTP only in the cytoskeleton-associated fraction. As a consequence of its phosphorylation, LMW PTP increases its catalytic activity about 20-fold. In this study, our interest was to investigate the role of LMW-PTP phosphorylation in cellular response to PDGF stimulation. To address this issue, we have transfected in NIH-3T3 cells a mutant form of LMW-PTP in which the c-Src phosphorylation sites (Tyr(131) and Tyr(132)) were mutated to alanine. We have established that LMW-PTP phosphorylation by c-Src after PDGF treatment strongly influences both cell adhesion and migration. In addition, we have discovered a new LMW-PTP substrate localized in the cytoskeleton that becomes tyrosine-phosphorylated after PDGF treatment: p190Rho-GAP. Hence, LMW-PTP plays multiple roles in PDGF receptor mediated mitogenesis, since it can bind and dephosphorylate PDGF receptor, and, at the same time, the cytoskeleton-associated LMW-PTP, through the regulation of the p190Rho-GAP phosphorylation state, controls the cytoskeleton rearrangement in response to PDGF stimulation. PMID- 10671493 TI - Potent growth inhibition of leukemic cells by novel ribbon-type antisense oligonucleotides to c-myb1. AB - We studied the effects of antisense oligonucleotides (AS oligos) with a novel structure. The AS oligos were covalently closed to avoid exonuclease activities by enzymatic ligation of two identical molecules. The AS oligos of a ribbon type (RiAS oligos) consist of two loops containing multiple antisense sequences and a stem connecting the two loops. Three antisense sequences targeting different binding sites were placed in a loop that was designed to form a minimal secondary structure by itself. RiAS oligos were found to be stable because they largely preserved their structural integrity after 24 h incubation in the presence of either exonuclease III or serums. When a human promyelocytic cell line, HL-60, was treated with RiAS oligos to c-myb, c-myb expression was effectively ablated. Cell growth was inhibited by >90% determined by both the 3-[4,5-dimethythiazol-2 yl]-2,5-diphenyltetrazolium bromide assay and [(3)H]thymidine incorporation. Further, when the leukemic cell line K562 was treated with c-myb RiAS oligos, colony formation on soft agarose was reduced by 92 +/- 2%. These results suggest that RiAS oligos may be employed for developing molecular antisense drugs as well as for the functional study of a gene. PMID- 10671494 TI - Inhibition of the Staphylococcus aureus NADPH-dependent enoyl-acyl carrier protein reductase by triclosan and hexachlorophene. AB - Enoyl-acyl carrier protein reductase (FabI) plays a determinant role in completing cycles of elongation in type II fatty acid synthase systems and is an important target for antibacterial drugs. The FabI component of Staphylococcus aureus (saFabI) was identified, and its properties were compared with Escherichia coli FabI (ecFabI). ecFabI and saFabI had similar specific activities, and saFabI expression complemented the E. coli fabI(Ts) mutant, illustrating that the Gram positive FabI was interchangeable with the Gram-negative FabI enzyme. However, ecFabI was specific for NADH, whereas saFabI exhibited specific and positive cooperative binding of NADPH. Triclosan and hexachlorophene inhibited both ecFabI and saFabI. The triclosan-resistant ecFabI(G93V) protein was also refractory to hexachlorophene inhibition, illustrating that both drugs bind at the FabI active site. Both the introduction of a plasmid expressing the safabI gene or a missense mutation in the chromosomal safabI gene led to triclosan resistance in S. aureus; however, these strains did not exhibit cross-resistance to hexachlorophene. The replacement of the ether linkage in triclosan by a carbon bridge in hexachlorophene prevented the formation of a stable FabI-NAD(P)(+)-drug ternary complex. Thus, the formation of this ternary complex is a key determinant of the antibacterial activity of FabI inhibitors. PMID- 10671495 TI - Essential role of phosphoinositide 3-kinase in leptin-induced K(ATP) channel activation in the rat CRI-G1 insulinoma cell line. AB - The mechanism by which leptin increases ATP-sensitive K(+) (K(ATP)) channel activity was investigated using the insulin-secreting cell line, CRI-G1. Wortmannin and LY 294002, inhibitors of phosphoinositide 3-kinase (PI3-kinase), prevented activation of K(ATP) channels by leptin. The inositol phospholipids phosphatidylinositol bisphosphate and phosphatidylinositol trisphosphate (PtdIns(3,4,5)P(3)) mimicked the effect of leptin by increasing K(ATP) channel activity in whole-cell and inside-out current recordings. LY 294002 prevented phosphatidylinositol bisphosphate, but not PtdIns(3,4,5)P(3), from increasing K(ATP) channel activity, consistent with the latter lipid acting as a membrane associated messenger linking leptin receptor activation and K(ATP) channels. Signaling cascades, activated downstream from PI 3-kinase, utilizing PtdIns(3,4,5)P(3) as a second messenger and commonly associated with insulin and cytokine action (MAPK, p70 ribosomal protein-S6 kinase, stress-activated protein kinase 2, p38 MAPK, and protein kinase B), do not appear to be involved in leptin mediated activation of K(ATP) channels in this cell line. Although PtdIns(3,4,5)P(3) appears a plausible and attractive candidate for the messenger that couples K(ATP) channels to leptin receptor activation, direct measurement of PtdIns(3,4,5)P(3) demonstrated that insulin, but not leptin, increased global cellular levels of PtdIns(3,4,5)P(3). Possible mechanisms to explain the involvement of PI 3-kinases in K(ATP) channel regulation are discussed. PMID- 10671496 TI - Identification of two major sites in the type I interleukin-1 receptor cytoplasmic region responsible for coupling to pro-inflammatory signaling pathways. AB - Type I interleukin-1 receptor is the prototype for a family of proteins, which play a central role in early responses to injury and infection. The similarity of function across the family is reflected in similarity in signaling: all members tested couple to activation of NFkappaB and stress kinases. The coupling to these pathways is mediated by a 200-residue intracellular domain (the Toll/interleukin 1 receptor domain), in which sequence conservation is primarily confined to three short motifs (boxes 1, 2, and 3) located at amino acid residue positions 10 (box 1), 60 (box 2), and 170 (box 3). We have analyzed the contribution of these motifs to function by alanine scanning mutagenesis of the human interleukin-1 receptor type I. Mutant receptors were tested for expression, ligand binding, activation of receptor-associated kinase(s), NFkappaB, stress kinases, and transcription. Mutations in all three motifs led to low cell surface expression. Mutants in box 3 were, however, wild type for signaling, whereas mutants in boxes 1 and 2 were defective. We conclude that the conserved motifs box 1 and box 2 mediate the coupling of molecules in the family to inflammation signaling pathways. PMID- 10671497 TI - Regulation of cytochrome bd expression in the obligate aerobe Azotobacter vinelandii by CydR (Fnr). Sensitivity to oxygen, reactive oxygen species, and nitric oxide. AB - Azotobacter vinelandii is an obligately aerobic bacterium in which aerotolerant nitrogen fixation requires cytochrome bd. Regulation of cytochrome bd expression is achieved by CydR (an Fnr homologue), which represses transcription of the oxidase genes cydAB. cydAB mRNA was mapped by primer extension; the transcriptional start site was determined, and putative -10 and -35 regions were deduced. Two "CydR boxes," one at the +1 site and one upstream of the -35 region, were identified. Transcriptionally inactive, purified CydR was converted, by adding NifS, cysteine, and Fe(2+), into an active form possessing acid-labile sulfide and spectra suggesting a [4Fe-4S](2+) cluster. Reconstituted CydR specifically bound both CydR boxes cooperatively, with higher affinity for the nearer consensus +1 site. Low concentrations of O(2) or NO ([O(2)]/[[CydR] or [NO]/[CydR] = 0.1-0. 6) elicited loss of the 420 nm absorbance attributed to the [4Fe-4S](2+) cluster, formation of a 315 nm species, and loss of ability to retard DNA migration. Retardation by reconstituted CydR was enhanced by superoxide dismutase and/or catalase, suggesting a role for reactive oxygen species in CydR inactivation. The role of CydR in regulating cydAB expression in the supposedly anoxic cytoplasm of A. vinelandii and similarities to cydAB regulation by Fnr in Escherichia coli are discussed. PMID- 10671498 TI - Molecular aspects of complement-mediated bacterial killing. Periplasmic conversion of C9 from a protoxin to a toxin. AB - As part of the membrane attack complex complement protein C9 is responsible for direct killing of bacteria. Here we show that in the periplasmic space of an Escherichia coli cell C9 is converted from a protoxin to a toxin by periplasmic conditions missing in spheroplasts. This conversion is independent of the pathway by which C9 enters the periplasm. Both, C9 shocked into the periplasm and plasmid expressed C9 targeted to the periplasm via a signal sequence are toxic. Toxicity requires disulfide-linked C9 because export into the periplasm of cells defective in disulfide bond synthesis (dsbA and dsbB mutants) is not toxic unless N acetylcysteine is added externally to promote cystines. A N-terminal fragment, C9[1-144], is not toxic nor is cytoplasmically expressed C9, even in trxB mutants that are able to form disulfide bonds in the cytoplasm. Importantly, expression of full-length C9 in complement-resistant cells has no effect on their viability. Expression and translocation into the periplasm may provide a novel model to identify molecular mechanisms of other bactericidal disulfide-linked proteins and to investigate the nature of bacterial complement resistance. PMID- 10671499 TI - Gbetagamma-dependent phosphoinositide 3-kinase activation in hearts with in vivo pressure overload hypertrophy. AB - Activation of phosphoinositide 3-kinases is coupled to both phosphotyrosine/growth factor and G protein-coupled receptors. We explored the role of phosphoinositide 3-kinase activation in myocardium during in vivo pressure overload hypertrophy in mice. Cytosolic extracts from wild type hypertrophied hearts showed a selective increase in the phosphoinositide 3-kinase gamma isoform. To address the role of G protein-coupled receptor-mediated activation of phosphoinositide 3-kinase, we used transgenic mice with cardiac specific overexpression of a Gbetagamma sequestering peptide. Extracts from hypertrophied transgenic hearts showed complete loss of phosphoinositide 3-kinase activation, indicating a Gbetagamma-dependent process. To determine the class of G proteins that contribute Gbetagamma dimers for in vivo phosphoinositide 3 kinase activation, two strategies were used: 1) transgenic mice with cardiac specific overexpression of a G(q) inhibitor peptide and 2) pertussis toxin treatment prior to pressure overload in wild type mice. Pressure overloaded G(q) inhibitor transgenic mice showed a complete absence of phosphoinositide 3-kinase activation, whereas pretreatment with pertussis toxin showed robust phosphoinositide 3-kinase activation. Taken together, these data demonstrate that activation of the phosphoinositide 3-kinase during in vivo pressure overload hypertrophy is Gbetagamma-dependent and the Gbetagamma dimers arise from stimulation of G(q)-coupled receptors. PMID- 10671500 TI - Evidence for intersubunit communication during acetyl-CoA cleavage by the multienzyme CO dehydrogenase/acetyl-CoA synthase complex from Methanosarcina thermophila. Evidence that the beta subunit catalyzes C-C and C-S bond cleavage. AB - The carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) from Methanosarcina thermophila is part of a five-subunit complex consisting of alpha, beta, gamma, delta, and epsilon subunits. The multienzyme complex catalyzes the reversible oxidation of CO to CO(2), transfer of the methyl group of acetyl-CoA to tetrahydromethanopterin (H(4)MPT), and acetyl-CoA synthesis from CO, CoA, and methyl-H(4)MPT. The alpha and epsilon subunits are required for CO oxidation. The gamma and delta subunits constitute a corrinoid iron-sulfur protein that is involved in the transmethylation reaction. This work focuses on the beta subunit. The isolated beta subunit contains significant amounts of nickel. When proteases truncate the beta subunit, causing the CODH/ACS complex to dissociate, the amount of intact beta subunit correlates directly with the EPR signal intensity of Cluster A and the activity of the CO/acetyl-CoA exchange reaction. Our results strongly indicate that the beta subunit harbors Cluster A, a NiFeS cluster, that is the active site of acetyl-CoA cleavage and assembly. Although the beta subunit is necessary, it is not sufficient for acetyl-CoA synthesis; interactions between the CODH and the ACS subunits are required for cleavage or synthesis of the C-C bond of acetyl-CoA. We propose that these interactions include intramolecular electron transfer reactions between the CODH and ACS subunits. PMID- 10671501 TI - LexA repressor forms stable dimers in solution. The role of specific dna in tightening protein-protein interactions. AB - Cooperativity in the interactions among proteins subunits and DNA is crucial for DNA recognition. LexA repressor was originally thought to bind DNA as a monomer, with cooperativity leading to tighter binding of the second monomer. The main support for this model was a high value of the dissociation constant for the LexA dimer (micromolar range). Here we show that the protein is a dimer at nanomolar concentrations under different conditions. The reversible dissociation of LexA dimer was investigated by the effects of hydrostatic pressure or urea, using fluorescence emission and polarization to monitor the dissociation process. The dissociation constant lies in the picomolar range (lower than 20 pM). LexA monomers associate with an unusual large volume change (340 ml/mol), indicating the burial of a large surface area upon dimerization. Whereas nonspecific DNA has no stabilizing effect, specific DNA induces tightening of the dimer and a 750 fold decrease in the K(d). In contrast to the previous model, a tight dimer rather than a monomer is the functional repressor. Accordingly, the LexA dimer only loses its ability to recognize a specific DNA sequence by RecA-induced autoproteolysis. Our work provides insights into the linkage between protein protein interactions, DNA recognition, and DNA repair. PMID- 10671502 TI - Enhancement of (45)Ca(2+) influx and voltage-dependent Ca(2+) channel activity by beta-amyloid-(1-40) in rat cortical synaptosomes and cultured cortical neurons. Modulation by the proinflammatory cytokine interleukin-1beta. AB - Beta-amyloid protein is thought to underlie the neurodegeneration associated with Alzheimer's disease by inducing Ca(2+)-dependent apoptosis. Elevated neuronal expression of the proinflammatory cytokine interleukin-1beta is an additional feature of neurodegeneration, and in this study we demonstrate that interleukin 1beta modulates the effects of beta-amyloid on Ca(2+) homeostasis in the rat cortex. beta-Amyloid-(1-40) (1 microM) caused a significant increase in (45)Ca(2+) influx into rat cortical synaptosomes via activation of L- and N-type voltage-dependent Ca(2+) channels and also increased the amplitude of N- and P type Ca(2+) channel currents recorded from cultured cortical neurons. In contrast, interleukin-1beta (5 ng/ml) reduced the (45)Ca(2+) influx into cortical synaptosomes and inhibited Ca(2+) channel activity in cultured cortical neurons. Furthermore, the stimulatory effects of beta-amyloid protein on Ca(2+) influx were blocked following exposure to interleukin-1beta, suggesting that interleukin 1beta may govern neuronal responses to beta-amyloid by regulating Ca(2+) homeostasis. PMID- 10671503 TI - Inhibition of the RelA(p65) NF-kappaB subunit by Egr-1. AB - Induction of transcription from the human immunodeficiency virus 1 long terminal repeat by the RelA (p65) NF-kappaB subunit has been shown to be dependent upon an interaction with the zinc finger DNA-binding domain of Sp1. It was unknown, however, whether NF-kappaB could also interact with other zinc finger-containing transcription factors. In this study we demonstrate that the early growth response transcription factor Egr-1, whose DNA-binding domain shares a high degree of homology with that of Sp1, can also interact with RelA in vitro and regulate NF-kappaB transcriptional activity in vivo. Similar to the interaction with Sp1, the Rel homology domain of RelA interacts with the zinc finger domain of Egr-1. Surprisingly, and in contrast to Sp1, Egr-1 specifically represses RelA transcriptional activity through its zinc finger domain. Moreover, the interaction between RelA and the Egr-1 zinc fingers is mutually exclusive with DNA binding suggesting a model in which Egr-1 directly sequesters NF-kappaB from its target promoters. Because Egr-1 is induced by many of the same stimuli that activate NF-kappaB, this novel transcriptional regulatory mechanism has many implications for the involvement of both factors in cellular processes such as apoptosis and the response to stress and infection. PMID- 10671504 TI - Different sterol regulatory element-binding protein-1 isoforms utilize distinct co-regulatory factors to activate the promoter for fatty acid synthase. AB - Sterol regulatory element-binding proteins (SREBPs) activate genes of cholesterol and fatty acid metabolism. In each case, a ubiquitous co-regulatory factor that binds to a neighboring recognition site is also required for efficient promoter activation. It is likely that gene- and pathway-specific regulation by the separate SREBP isoforms is dependent on subtle differences in how the individual proteins function with specific co-regulators to activate gene expression. In the studies reported here we extend these observations significantly by demonstrating that SREBPs are involved in both sterol regulation and carbohydrate activation of the FAS promoter. We also demonstrate that the previously implicated Sp1 site is largely dispensable for sterol regulation in established cultured cells, whereas a CCAAT-binding factor/nuclear factor Y is critically important. In contrast, carbohydrate activation of the FAS promoter in primary hepatocytes is dependent upon SREBP and both the Sp1 and CCAAT-binding factor/nuclear factor Y sites. Because 1c is the predominant SREBP isoform expressed in hepatocytes and 1a is more abundant in sterol depleted established cell lines, this suggests that the different SREBP isoforms utilize distinct co-regulatory factors to activate target gene expression. PMID- 10671505 TI - The monoclonal antibody 1F6 identifies a pH-dependent conformational change in the hydrophilic NH(2) terminus of NhaA Na(+)/H(+) antiporter of Escherichia coli. AB - One of the most interesting properties of the NhaA Na(+)/H(+) antiporter of Escherichia coli is the strong regulation of its activity by pH. This regulation is accompanied by a conformational change that can be probed by digestion with trypsin and involves the hydrophilic loop connecting the transmembrane helices VIII-IX. In the present work we show that a monoclonal antibody (mAb), 1F6, recognizes yet another domain of NhaA in a pH-dependent manner. This antibody binds NhaA at pH 8.5 but not at pH 4.5, whereas two other mAbs bind to NhaA independently of pH. The epitope of mAb 1F6 was located at the NH(2) terminus of NhaA by probing proteolytic fragments in Western blot analysis and amino acid sequencing. The antibody bound to the peptide HLHRFFSS, starting at the third amino acid of NhaA. A synthetic peptide with this sequence was shown to bind mAb 1F6 both at acidic and alkaline pH suggesting that this peptide is accessible to mAb 1F6 in the native protein only at alkaline pH. Although slightly shifted to acidic pH, the pH profile of the binding of mAb 1F6 to the antiporter is similar to that of both the Na(+)/H(+) antiporter activity as well as to its sensitivity to trypsin. We thus suggest that these pH profiles reflect a pH-dependent conformational change, which leads to activation of the antiporter. Indeed, a replacement of Gly-338 by Ser (G338S), which alleviates the pH dependence of both the NhaA activity as well as its sensitivity to trypsin, affects in a similar pattern the binding of mAb 1F6 to NhaA. Furthermore, the binding site of mAb 1F6 is involved in the functioning of the antiporter as follows: a double Cys replacement H3C/H5C causes an acidic shift by half a pH unit in the pH dependence of the antiporter; N-ethylmaleimide, which does not inhibit the wild-type protein, inhibits H3C/H5C antiporter to an extent similar to that exerted by mAb 1F6. PMID- 10671506 TI - Stereospecificity of hydrogen abstraction in the conversion of arachidonic acid to 15R-HETE by aspirin-treated cyclooxygenase-2. Implications for the alignment of substrate in the active site. AB - The initial and rate-limiting step in prostaglandin biosynthesis is stereoselective removal of the pro-S hydrogen from the 13-carbon of arachidonic acid. This is followed by oxygenation at C-11, formation of the five-membered ring, and a second oxygenation at C-15 to yield the endoperoxide product, prostaglandin G(2). Aspirin treatment of cyclooxygenase-2 is known to acetylate an active site serine, block prostaglandin biosynthesis, and give 15R hydroxyeicosatetraenoic acid (15R-HETE) as the only product. 15R-HETE and prostaglandins have opposite stereoconfigurations of the 15-hydroxyl. To understand the changes that lead to 15R-HETE synthesis in aspirin-treated COX-2, we employed pro-R- and pro-S-labeled [13-(3)H]arachidonic acids to investigate the selectivity of the initial hydrogen abstraction. Remarkably, aspirin-treated COX-2 formed 15R-HETE with removal of the pro-S hydrogen at C-13 (3-9% retention of pro-S tritium label), the same stereoselectivity as in the formation of prostaglandins by native cyclooxygenase. To account for this result and the change in oxygenase specificity, we suggest that the bulky serine acetyl group forces a realignment of the omega end of the arachidonic acid carbon chain. This can rationalize abstraction of the C-13 pro-S hydrogen, the blocking of prostaglandin synthesis, and the formation of 15R-HETE as the sole enzymatic product. PMID- 10671507 TI - Characterization of chimeric lipopolysaccharides from Escherichia coli strain JM109 transformed with lipooligosaccharide synthesis genes (lsg) from Haemophilus influenzae. AB - Previously, we reported the expression of chimeric lipopolysaccharides (LPS) in Escherichia coli strain JM109 (a K-12 strain) transformed with plasmids containing Haemophilus influenzae lipooligosaccharide synthesis genes (lsg) (Abu Kwaik, Y., McLaughlin, R. E., Apicella, M. A., and Spinola, S. M. (1991) Mol. Microbiol. 5, 2475-2480). In this current study, we have analyzed the O deacylated LPS and free oligosaccharides from three transformants (designated pGEMLOS-4, pGEMLOS-5, and pGEMLOS-7) by matrix-assisted laser desorption ionization, electrospray ionization, and tandem mass spectrometry techniques, along with composition and linkage analyses. These data show that the chimeric LPS consist of the complete E. coli LPS core structure glycosylated on the 7 position of the non-reducing terminal branch heptose with oligosaccharides from H. influenzae. In pGEMLOS-7, the disaccharide Gal1--> 3GlcNAc1--> is added, and in pGEMLOS-5, the structure is extended to Gal1-->4GlcNAc1-->3Gal1-->3GlcNAc1-->. PGEMLOS-5 LPS reacts positively with monoclonal antibody 3F11, an antibody that recognizes the terminal disaccharide of lacto-N-neotetraose. In pGEMLOS-4 LPS, the 3F11 epitope is apparently blocked by glycosylation on the 6-position of the terminal Gal with either Gal or GlcNAc. The biosynthesis of these chimeric LPS was found to be dependent on a functional wecA (formerly rfe) gene in E. coli. By using this carbohydrate expression system, we have been able to examine the functions of the lsg genes independent of the effects of other endogenous Haemophilus genes and expressed proteins. PMID- 10671508 TI - Transport and processing of endogenously synthesized ApoE on the macrophage cell surface. AB - We have previously established the presence of a pool of apoE sequestered on the macrophage cell surface by demonstrating its displacement from a cell monolayer at 4 degrees C. In this series of experiments, we use a cell surface biotinylation protocol to directly quantitate apoE on the macrophage cell surface and evaluate its transport to and from this cell surface pool. In human monocyte derived macrophages labeled to equilibrium and in a mouse macrophage cell line transfected to constitutively express human apoE3, approximately 8% of total cellular apoE was present on the surface, but only a portion of this surface pool served as a direct precursor to secreted apoE. The half-life of apoE on the macrophage cell surface was calculated to be approximately 12 min. On SDS polyacrylamide gel electrophoresis, the apoE isolated from the surface fraction of cells labeled to equilibrium migrated in an isoform pattern distinct from that observed from the intracellular fraction, with the surface fraction migrating predominantly in a higher molecular weight isoform. Pulse labeling experiments demonstrated that newly synthesized apoE reached the cell surface by 10 min but was predominantly in a low molecular weight isoform. There was also a lag between appearance of apoE on the cell surface and its appearance in the medium. Biotinylated apoE, which accumulated in the medium, even from pulse labeled cells, was predominantly in the high molecular weight isoform. Additional experiments demonstrated that low molecular weight apoE present on the cell surface was modified to higher molecular weight apoE by the addition of sialic acid residues prior to secretion and that this conversion was inhibited by brefeldin A. These results demonstrate an unexpected complexity in the transport and cellular processing of macrophage cell surface apoE. Factors that modulate the size and turnover of the cell surface pool of apoE in the macrophage remain to be identified and investigated. PMID- 10671509 TI - Oxidized low density lipoproteins regulate synthesis of monkey aortic smooth muscle cell proteoglycans that have enhanced native low density lipoprotein binding properties. AB - Oxidized low density lipoproteins (Ox-LDL) affect several biological processes involved in atherogenesis. However, it is not known whether Ox-LDL can regulate proteoglycan expression and thus affect arterial wall lipoprotein retention. This study evaluated whether Ox-LDL, as compared with native LDL, regulates proteoglycan expression by monkey arterial smooth muscle cells in vitro and whether proteoglycans synthesized in the presence of Ox-LDL exhibit altered lipoprotein binding properties. Ox-LDL stimulated glycosaminoglycan synthesis, as measured by (35)SO(4) incorporation, by 30-50% over that of native LDL. The effect was maximal after 72 h of exposure to 5 microg/ml of Ox-LDL. The molecular sizes of versican, biglycan, and decorin increased in response to Ox-LDL, as indicated by size exclusion chromatography and SDS-polyacrylamide gel electrophoresis. These effects could be mimicked by the lipid extract of Ox-LDL. These size increases were largely due to chain elongation and not to alterations in the ratio of (35)SO(4) to [(3)H]glucosamine incorporation. Affinity chromatography indicated that Ox-LDL stimulated the synthesis of proteoglycans with high affinity for native LDL. Ox-LDL also specifically stimulated mRNA expression for biglycan (but not versican or decorin), which was correlated with increased expression of secreted biglycan. Thus, Ox-LDL may influence lipoprotein retention by regulating synthesis of biglycan and also by altering glycosaminoglycan synthesis of vascular proteoglycans so as to enhance lipoprotein binding properties. PMID- 10671510 TI - Protein stability and domain topology determine the transcriptional activity of the mammalian glial cells missing homolog, GCMb. AB - The glial cells missing (GCM) family of transcription factors consists of Drosophila GCM and the mammalian proteins GCMa and GCMb. They are expressed in a highly restricted manner during development and are known or assumed to be important regulators of developmental fate decisions. As the biochemical properties of GCMb have not been studied so far, we have undertaken a detailed structure-function analysis of the mouse GCMb (mGCMb) protein. DNA-binding specificity was very similar to that of other GCM proteins. Nevertheless, mGCMb was only a weak transcriptional activator in a number of different tissue culture systems. Interestingly, this was not due to an intrinsic absence of transactivation potential. In effect, we were able to identify two separate transactivation domains within mGCMb, one carboxyl-terminally adjacent to the DNA binding domain and the second within the extreme carboxyl terminus. Activity of both transactivation domains was, however, modulated by an inhibitory region unique to mGCMb and located between the two transactivation domains. Furthermore, pulse-chase experiments proved that the mGCMb protein has a half-life approximately four times shorter than mGCMa. Introduction of the above mentioned inhibitory domain of mGCMb into mGCMa shortened the half-life of mGCMa to a value typical of mGCMb with a concomitant reduction in transactivation potential. Given the strong correlation between protein stability and transactivation potential, functional differences between the two mammalian GCM homologs are likely due to differences in stability with a single inhibitory region in mGCMb being involved in the reduction of both. PMID- 10671511 TI - Group V phospholipase A(2)-mediated oleic acid mobilization in lipopolysaccharide stimulated P388D(1) macrophages. AB - P388D(1) macrophages prelabeled with [(3)H]arachidonic acid (AA) respond to bacterial lipopolysaccharide (LPS) by mobilizing AA in a process that takes several hours and is mediated by the concerted actions of the group IV cytosolic phospholipase A(2) and the group V secretory phospholipase A(2) (sPLA(2)). Here we show that when the LPS-activated cells are prelabeled with [(3)H]oleic acid (OA), they also mobilize and release OA to the extracellular medium. The time and concentration dependence of the LPS effect on OA release fully resemble those of the AA release. Experiments in which both AA and OA release are measured simultaneously indicate that AA is released 3 times more efficiently than OA. Importantly, LPS-stimulated OA release is strongly inhibited by the selective sPLA(2) inhibitors 3-(3-acetamide-1-benzyl-2-ethylindolyl-5-oxy)propane sulfonic acid and carboxymethylcellulose-linked phosphatidylethanolamine. The addition of exogenous recombinant sPLA(2) to the cells also triggers OA release. These data implicate a functionally active sPLA(2) as being essential for the cells to release OA upon stimulation with LPS. OA release is also inhibited by methyl arachidonyl fluorophosphonate but not by bromoenol lactone, indicating that the group IV cytosolic phospholipase A(2) is also involved in the process. Together, these data reveal that OA release occurs during stimulation of the P388D(1) macrophages by LPS and that the regulatory features of the OA release are strikingly similar to those previously found for the AA release. PMID- 10671512 TI - Characterization of the insulin-regulated endocytic recycling mechanism in 3T3-L1 adipocytes using a novel reporter molecule. AB - The endocytic trafficking of the GLUT4 glucose transporter and the insulin regulated aminopeptidase (IRAP) are regulated by insulin. We have used a chimera between the intracellular domain of IRAP and the extracellular and transmembrane domains of the transferrin receptor (vpTR) to characterize IRAP-like trafficking in 3T3-L1 adipocytes. Our data demonstrate that the cytoplasmic domain of IRAP is sufficient to target vpTR to the insulin-regulated, slow recycling pathway in adipocytes and that the dynamic retention of vpTR is dependent on a di-leucine motif. Our kinetic analysis demonstrates that vpTR recycles as a single kinetic pool and that vpTR is very efficiently sorted from endosomes to the insulin regulated recycling pathway. An implication of these findings is that the key step in the dynamic retention of vpTR occurs within the early endosomal system. We have previously shown that vpTR is trafficked by an insulin-regulated pathway in Chinese hamster ovary cells (Johnson, A. O., Subtil, A., Petrush, R., Kobylarz, K., Keller, S., and Mc Graw, T. E. (1998) J. Biol. Chem. 273, 17968 17977). The behavior of vpTR in Chinese hamster ovary cells is similar to its behavior in 3T3-L1 adipocytes. The main difference is that insulin has a larger effect on the trafficking of vpTR in the adipocytes. We concluded that the insulin-regulated slow recycling endocytic mechanism is expressed in many different cell types and therefore is not a unique characteristic of cells that express GLUT4. PMID- 10671513 TI - Regulation of tumor cell chemotaxis by type IV collagen is mediated by a Ca(2+) dependent mechanism requiring CD47 and the integrin alpha(V)beta(3). AB - Studies from our laboratories demonstrated that synthetic peptides from the non collagenous (NC-1) domain of the alpha3 (IV) chain of type IV collagen (COL IV) enhanced tumor cell adhesion (Han, J., Ohno, N., Monboisse, J. C., Pasco, S., Borel, J. P., and Kefalides, N. A. (1997) J. Biol. Chem. 272, 20395-20401). We have isolated the receptors for the alpha3(IV)185-203 peptide from melanoma and prostate tumor cells and identified them as CD47/integrin-associated protein and the integrin alpha(V)beta(3) (Shahan, T. A., Ziaie, Z., Pasco, S., Fawzi, A., Bellon, G., Monboisse, J. C., and Kefalides, N. A. (1999) Cancer Res. 59, 4584 4590). In the present study we have examined the effect of CD47 and the integrin alpha(V)beta(3) on in vitro tumor cell chemotaxis and Ca(2+)(i) modulation in response to COL IV, from the anterior lens capsule (ALC-COL IV) and peptides from its NC-1 domain. COL IV as well as the alpha3(IV) peptide promoted tumor cell chemotaxis with an immediate increase in intracellular [Ca(2+)]. Treating tumor cells with CD47 and integrin alpha(V)beta(3)-reactive antibodies reduced chemotaxis as well as the rise in [Ca(2+)](i) in response to ALC-COL IV or the alpha3(IV)185-203 peptide but not to Engelbreth-Holm-Swarm-COL IV or fibronectin. The alpha3(IV)185-203 synthetic peptide stimulated an increase in calcium from intracellular stores exclusively, whereas ALC-COL IV, Engelbreth-Holm-Swarm-COL IV, and fibronectin stimulated Ca(2+) flux from both internal and external stores. Furthermore, treatment of the cells with Ca(2+) chelator bis-(O aminophenoxyl)ethane-N,N,N',N'-tetraaceticacid- acetomethoxy ester inhibited chemotaxis toward both ALC-COL IV and the alpha3(IV)185-203 peptide. These data indicate that CD47 and integrin alpha(V)beta(3) regulate tumor cell chemotaxis in response to COL IV and the alpha3(IV)185-203 peptide through a Ca(2+)-dependent mechanism. PMID- 10671514 TI - Differential regulation of the phosphatidylinositol 3-kinase/Akt and p70 S6 kinase pathways by the alpha(1A)-adrenergic receptor in rat-1 fibroblasts. AB - Phosphatidylinositol (PI) 3-kinase and its downstream effector Akt are thought to be signaling intermediates that link cell surface receptors to p70 S6 kinase. We examined the effect of a G(q)-coupled receptor on PI 3-kinase/Akt signaling and p70 S6 kinase activation using Rat-1 fibroblasts stably expressing the human alpha(1A)-adrenergic receptor. Treatment of the cells with phenylephrine, a specific alpha(1)-adrenergic receptor agonist, activated p70 S6 kinase but did not activate PI 3-kinase or any of the three known isoforms of Akt. Furthermore, phenylephrine blocked the insulin-like growth factor-I (IGF-I)-induced activation of PI 3-kinase and the phosphorylation and activation of Akt-1. The effect of phenylephrine was not confined to signaling pathways that include insulin receptor substrate-1, as the alpha(1)-adrenergic receptor agonist also inhibited the platelet-derived growth factor-induced activation of PI 3-kinase and Akt-1. Although increasing the intracellular Ca(2+) concentration with the ionophore A23187 inhibited the activation of Akt-1 by IGF-I, Ca(2+) does not appear to play a role in the phenylephrine-mediated inhibition of the PI 3-kinase/Akt pathway. The differential ability of phenylephrine and IGF-I to activate Akt-1 resulted in a differential ability to protect cells from UV-induced apoptosis. These results demonstrate that activation of p70 S6 kinase by the alpha(1A)-adrenergic receptor in Rat-1 fibroblasts occurs in the absence of PI 3-kinase/Akt signaling. Furthermore, this receptor negatively regulates the PI 3-kinase/Akt pathway, resulting in enhanced cell death following apoptotic insult. PMID- 10671515 TI - A hemoglobin with an optical function. AB - Hemoglobins are best known as oxygen transport proteins. Here we describe a hemoglobin from the parasitic nematode Mermis nigrescens (Mn-GLB-E) that has an optical, light shadowing function. The protein accumulates to high concentration as intracellular crystals in the ocellus of mature phototactic adult females while also being expressed at low concentration in other tissues. It differs in sequence and expression pattern from Mn-GLB-B, a second Mermis globin. It retains the structure and oxygen-binding and light-absorbing properties typical of nematode hemoglobins. As such, recruitment to a shadowing role in the eye appears to have occurred by changes in expression without modification of biochemistry. Both globins are coded by genes interrupted by two introns at the conserved positions B12.2 and G7.0, which is in agreement with the 3exon/2intron pattern model of globin gene evolution. PMID- 10671516 TI - Proton delivery in NO reduction by fungal nitric-oxide reductase. Cryogenic crystallography, spectroscopy, and kinetics of ferric-NO complexes of wild-type and mutant enzymes. AB - Fungal nitric-oxide reductase (NOR) is a heme enzyme that catalyzes the reduction of NO to N(2)O through its ferric-NO complex, the first intermediate of the catalysis. Crystal structures of the ferric-NO forms of wild type (WT) fungal NOR, and of the Ser(286) --> Val and Ser(286) --> Thr mutant enzymes were determined to 1.7-A resolution at cryogenic temperature (100 K). This shows a slightly tilted and bent NO binding to the heme iron, in sharp contrast to the highly bent NO coordination found in ferrous hemoproteins. In the WT structure, a specific hydrogen-bonding network that connects the active site to the solvent was identified, H(2)O(Wat(74))-Ser(286)-H(2)O(Wat(33))-Asp(393)-solvent. Wat(74) is located 3.10 A from the iron-bound NO. Replacement of Ser(286) with Val or Thr scarcely alters the NO coordination structure but expels the water molecules, Wat(74) from the active site. The Asp(393) mutation does not influence the position of Wat(74), but disrupts the hydrogen-bonding network at Wat(33), as evidenced by enzymatic, kinetic, and spectroscopic (resonance Raman and IR) results. The structural changes observed upon the Ser(286) or the Asp(393) mutation are consistent with the dramatic loss of the enzymatic activity for the NO reduction of fungal NOR. We have conclusively identified the water molecule, Wat(74), adjacent to the iron-bound NO as a proton donor to the Fe-NO moiety. In addition, we find the hydrogen-bonding network, H(2)O(Wat(74))-Ser(286) H(2)O(Wat(33))-Asp(393), as a proton delivery pathway in the NO reduction reaction by fungal NOR. PMID- 10671517 TI - ERO1-L, a human protein that favors disulfide bond formation in the endoplasmic reticulum. AB - Oxidizing conditions must be maintained in the endoplasmic reticulum (ER) to allow the formation of disulfide bonds in secretory proteins. Here we report the cloning and characterization of a mammalian gene (ERO1-L) that shares extensive homology with the Saccharomyces cerevisiae ERO1 gene, required in yeast for oxidative protein folding. When expressed in mammalian cells, the product of the human ERO1-L gene co-localizes with ER markers and displays Endo-H-sensitive glycans. In isolated microsomes, ERO1-L behaves as a type II integral membrane protein. ERO1-L is able to complement several phenotypic traits of the yeast thermosensitive mutant ero1-1, including temperature and dithiothreitol sensitivity, and intrachain disulfide bond formation in carboxypeptidase Y. ERO1 L is no longer functional when either one of the highly conserved Cys-394 or Cys 397 is mutated. These results strongly suggest that ERO1-L is involved in oxidative ER protein folding in mammalian cells. PMID- 10671518 TI - A dynactin subunit with a highly conserved cysteine-rich motif interacts directly with Arp1. AB - Dynactin is a multisubunit complex and a required cofactor for most, or all, of the cellular processes powered by the microtubule-based motor cytoplasmic dynein. Using a dynein affinity column, the previously uncharacterized p62 subunit of dynactin was isolated and microsequenced. Two peptide sequences were used to clone human cDNAs encoding p62. Sequence analysis of the predicted human polypeptide of 53 kDa revealed a highly conserved pattern of eleven cysteine residues, eight of which fit the consensus sequence for a Zn(2+)-binding RING domain. We have characterized p62 as an integral component of 20 S dynactin by biochemical and immunocytochemical methods. Affinity chromatography experiments demonstrate that p62 binds directly to the Arp1 subunit of dynactin. Immunocytochemistry with antibodies to p62 demonstrates that this polypeptide has a punctate cytoplasmic distribution as well as centrosomal distribution typical of dynactin. In transfected cells, overexpression of p62 did not disrupt microtubule organization or the integrity of the Golgi but did cause both cytosolic and nuclear distribution of the protein, suggesting that this polypeptide may be targeted to the nucleus at very high expression levels. PMID- 10671519 TI - MAN1, an inner nuclear membrane protein that shares the LEM domain with lamina associated polypeptide 2 and emerin. AB - The "MAN antigens" are polypeptides recognized by autoantibodies from a patient with a collagen vascular disease and localized to the nuclear envelope. We now show that one of the human MAN antigens termed MAN1 is a 82.3-kDa protein with an amino-terminal domain followed by two hydrophobic segments and a carboxyl terminal tail. The MAN1 gene contains seven protein-coding exons and is assigned to human chromosome 12q14. Its mRNA is approximately 5.5 kilobases and is detected in several different cell types that were examined. Cell extraction experiments show that MAN1 is an integral membrane protein. When expressed in transfected cells, MAN1 is exclusively targeted to the nuclear envelope, consistent with an inner nuclear membrane localization. Protein sequence analysis reveals that MAN1 shares a conserved globular domain of approximately 40 amino acids, which we term the LEM module, with inner nuclear membrane proteins lamina associated polypeptide 2 and emerin. The LEM module is also present in two proteins of Caenorhabditis elegans. These results show that MAN1 is an integral protein of the inner nuclear membrane that shares the LEM module with other proteins of this subcellular localization. PMID- 10671520 TI - Molecular cloning of a novel human I-mfa domain-containing protein that differently regulates human T-cell leukemia virus type I and HIV-1 expression. AB - Regulation of viral genome expression is the result of complex cooperation between viral proteins and host cell factors. We report here the characterization of a novel cellular factor sharing homology with the specific cysteine-rich C terminal domain of the basic helix-loop-helix repressor protein I-mfa. The synthesis of this new factor, called HIC for Human I-mfa domain-Containing protein, is controlled at the translational level by two different codons, an ATG and an upstream non-ATG translational initiator, allowing the production of two protein isoforms, p32 and p40, respectively. We show that the HIC protein isoforms present different subcellular localizations, p32 being mainly distributed throughout the cytoplasm, whereas p40 is targeted to the nucleolus. Moreover, in trying to understand the function of HIC, we have found that both isoforms stimulate in T-cells the expression of a luciferase reporter gene driven by the human T-cell leukemia virus type I-long terminal repeat in the presence of the viral transactivator Tax. We demonstrate by mutagenesis that the I-mfa-like domain of HIC is involved in this regulation. Finally, we also show that HIC is able to down-regulate the luciferase expression from the human immunodeficiency virus type 1-long terminal repeat induced by the viral transactivator Tat. From these results, we propose that HIC and I-mfa represent two members of a new family of proteins regulating gene expression and characterized by a particular cysteine-rich C-terminal domain. PMID- 10671521 TI - Tumor necrosis factor-alpha induces differentiation of and bone resorption by osteoclasts. AB - Osteoclast progenitors differentiate into mature osteoclasts in the presence of receptor activator of NF-kappaB (RANK) ligand on stromal or osteoblastic cells and monocyte macrophage colony-stimulating factor (M-CSF). The soluble RANK ligand induces the same differentiation in vitro without stromal cells. Tumor necrosis factor-alpha (TNF-alpha), a potent cytokine involved in the regulation of osteoclast activity, promotes bone resorption via a primary effect on osteoblasts; however, it remains unclear whether TNF-alpha can also directly induce the differentiation of osteoclast progenitors into mature osteoclasts. This study revealed that TNF-alpha directly induced the formation of tartrate resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs), which produced resorption pits on bone in vitro in the presence of M-CSF. The bone resorption activity of TNF-alpha-induced MNCs was lower than that of soluble RANK ligand-induced MNCs; however, interleukin-1beta stimulated this activity of TNF alpha-induced MNCs without an increase in the number of MNCs. In this case, interleukin-1beta did not induce TRAP-positive MNC formation. The osteoclast progenitors expressed TNF receptors, p55 and p75; and the induction of TRAP positive MNCs by TNF-alpha was inhibited completely by an anti-p55 antibody and partially by an anti-p75 antibody. Our findings presented here are the first to indicate that TNF-alpha is a crucial differentiation factor for osteoclasts. Our results suggest that TNF-alpha and M-CSF play an important role in local osteolysis in chronic inflammatory diseases. PMID- 10671522 TI - Carboxypeptidase Z is present in the regulated secretory pathway and extracellular matrix in cultured cells and in human tissues. AB - Carboxypeptidase Z (CPZ) is a newly reported member of the metallocarboxypeptidase gene family, but unlike other members of this family, CPZ contains an N-terminal domain that has amino acid sequence similarity to Wnt binding proteins. In order to gain insights as to the potential function of CPZ, the intracellular localization of this protein was determined in cell culture and in human tissues. When expressed in the AtT-20 mouse pituitary cell line, CPZ protein is routed to the regulated secretory pathway and secreted upon stimulation. A fraction of the secreted CPZ remains associated with the extracellular matrix. Endogenous CPZ in the PC12 rat pheochromocytoma cell line is also associated with the extracellular matrix. In human placenta, CPZ is present within invasive trophoblasts and in the surrounding extracellular space, indicating an association with extracellular matrix. CPZ is also present in amnion cells, but is not readily apparent in the extracellular matrix of this cell type. A human adenocarcinoma of the colon shows expression of CPZ in the extracellular matrix adjacent to malignant cells. Taken together, CPZ appears to be a component of the extracellular matrix in some cell types, where it may function in the binding of Wnt. PMID- 10671523 TI - The molecular structure of hyperthermostable aromatic aminotransferase with novel substrate specificity from Pyrococcus horikoshii. AB - Aromatic amino acid aminotransferase (ArATPh), which has a melting temperature of 120 degrees C, is one of the most thermostable aminotransferases yet to be discovered. The crystal structure of this aminotransferase from the hyperthermophilic archaeon Pyrococcus horikoshii was determined to a resolution of 2.1 A. ArATPh has a homodimer structure in which each subunit is composed of two domains, in a manner similar to other well characterized aminotransferases. By the least square fit after superposing on a mesophilic ArAT, the ArATPh molecule exhibits a large deviation of the main chain coordinates, three shortened alpha-helices, an elongated loop connecting two domains, and a long loop transformed from an alpha-helix, which are all factors that are likely to contribute to its hyperthermostability. The pyridine ring of the cofactor pyridoxal 5'-phosphate covalently binding to Lys(233) is stacked parallel to F121 on one side and interacts with the geminal dimethyl-CH/pi groups of Val(201) on the other side. This tight stacking against the pyridine ring probably contributes to the hyperthermostability of ArATPh. Compared with other ArATs, ArATPh has a novel substrate specificity, the order of preference being Tyr > Phe > Glu > Trp > His>> Met > Leu > Asp > Asn. Its relatively weak activity against Asp is due to lack of an arginine residue corresponding to Arg(292)* (where the asterisk indicates that this is a residues supplied by the other subunit of the dimer) in pig cytosolic aspartate aminotransferase. The enzyme recognizes the aromatic substrate by hydrophobic interaction with aromatic rings (Phe(121) and Tyr(59)*) and probably recognizes acidic substrates by a hydrophilic interaction involving a hydrogen bond network with Thr(264)*. PMID- 10671524 TI - Locking regulatory myosin in the off-state with trifluoperazine. AB - Scallop striated adductor muscle myosin is a regulatory myosin, its activity being controlled directly through calcium binding. Here, we show that millimolar concentrations of trifluoperazine were effective at removal of all regulatory light chains from scallop myosin or myofibrils. More important, 200 microM trifluoperazine, a concentration 10-fold less than that required for light-chain removal, resulted in the reversible elimination of actin-activated and intrinsic ATPase activities. Unlike desensitization induced by metal ion chelation, which leads to an elevation of activity in the absence of calcium concurrent with regulatory light-chain removal, trifluoperazine caused a decline in actin activated MgATPase activity both in the presence and absence of calcium. Procedures were equally effective with respect to scallop myosin, myofibrils, subfragment-1, or desensitized myofibrils. Increased alpha-helicity could be induced in the isolated essential light chain through addition of 100-200 microM trifluoperazine. We propose that micromolar concentrations of trifluoperazine disrupt regulation by binding to a single high-affinity site located in the C terminal domain of the essential light chain, which locks scallop myosin in a conformation resembling the off-state. At millimolar trifluoperazine concentrations, additional binding sites on both light chains would be filled, leading to regulatory light-chain displacement. PMID- 10671525 TI - Pairwise interactions between neuronal alpha(7) acetylcholine receptors and alpha conotoxin PnIB. AB - This work uses alpha-conotoxin PnIB to probe the agonist binding site of neuronal alpha(7) acetylcholine receptors. We mutated the 13 non-cysteine residues in CTx PnIB, expressed alpha(7)/5-hydroxytryptamine-3 homomeric receptors in 293 HEK cells, and measured binding of each mutant toxin to the expressed receptors by competition against the initial rate of (125)I-alpha-bungarotoxin binding. The results reveal that residues Ser-4, Leu-5, Pro-6, Pro-7, Ala-9, and Leu-10 endow CTx PnIB with affinity for alpha(7)/5-hydroxytryptamine-3 receptors; side chains of these residues cluster in a localized region within the three-dimensional structure of CTx PnIB. We next mutated key residues in the seven loops of alpha(7) that converge at subunit interfaces to form the agonist binding site. The results reveal predominant contributions by residues Trp-149 and Tyr-93 in alpha(7) and smaller contributions by Ser-34, Arg-186, Tyr-188, and Tyr-195. To identify pairwise interactions that stabilize the receptor-conotoxin complex, we measured binding of receptor and toxin mutations and analyzed the results by double mutant cycles. The results reveal a single dominant interaction between Leu-10 of CTx PnIB and Trp-149 of alpha(7) that anchors the toxin to the binding site. We also find weaker interactions between Pro-6 of CTx PnIB and Trp-149 and between both Pro-6 and Pro-7 and Tyr-93 of alpha(7). The overall results demonstrate that a localized hydrophobic region in CTx PnIB interacts with conserved aromatic residues on one of the two faces of the alpha(7) binding site. PMID- 10671526 TI - Binding and phosphorylation of a novel male germ cell-specific cGMP-dependent protein kinase-anchoring protein by cGMP-dependent protein kinase Ialpha. AB - cGMP-dependent protein kinase (cGK) is a major cellular receptor of cGMP and plays important roles in cGMP-dependent signal transduction pathways. To isolate the components of the cGMP/cGK signaling pathway such as substrates and regulatory proteins of cGK, we employed the yeast two-hybrid system using cGK Ialpha as a bait and isolated a novel male germ cell-specific 42-kDa protein, GKAP42 (42-kDa cGMP-dependent protein kinase anchoring protein). Although the N terminal region (amino acids 1-66) of cGK-Ialpha is sufficient for the association with GKAP42, GKAP42 could not interact with cGK-Ibeta, cGK-II, or cAMP-dependent protein kinase. GKAP42 mRNA is specifically expressed in testis, where it is restricted to the spermatocytes and early round spermatids. Endogenous cGK-I is co-immunoprecipitated with anti-GKAP42 antibody from mouse testis tissue, suggesting that cGK-I physiologically interacts with GKAP42. Immunocytochemical observations revealed that GKAP42 is localized to the Golgi complex and that cGK-Ialpha is co-localized to the Golgi complex when coexpressed with GKAP42. Although both cGK-Ialpha and -Ibeta, but not cAMP-dependent protein kinase, phosphorylated GKAP42 in vitro, GKAP42 was a good substrate only for cGK Ialpha in intact cells, suggesting that the association with kinase protein is required for the phosphorylation in vivo. Finally, we demonstrated that the kinase-deficient mutant of cGK-Ialpha stably associates with GKAP42 and that binding of cGMP to cGK-Ialpha facilitates their release from GKAP42. These findings suggest that GKAP42 functions as an anchoring protein for cGK-Ialpha and that cGK-Ialpha may participate in germ cell development through phosphorylation of Golgi-associated proteins such as GKAP42. PMID- 10671527 TI - A eukaryotic alanine racemase gene involved in cyclic peptide biosynthesis. AB - The cyclic tetrapeptide HC-toxin is an essential virulence determinant for the plant pathogenic fungus Cochliobolus carbonum and an inhibitor of histone deacetylase. The major form of HC-toxin contains the D-isomers of Ala and Pro. The non-ribosomal peptide synthetase that synthesizes HC-toxin has only one epimerizing domain for conversion of L-Pro to D-Pro; the source of D-Ala has remained unknown. Here we present the cloning and characterization of a new gene involved in HC-toxin biosynthesis, TOXG. TOXG is present only in HC-toxin producing (Tox2(+)) isolates of C. carbonum. TOXG is able to support D-Ala independent growth of a strain of Escherichia coli defective in D-Ala synthesis. A C. carbonum strain with both of its copies of TOXG mutated grows normally in culture, and although it no longer makes the three forms of HC-toxin that contain D-Ala, it still makes a minor form of HC-toxin that contains Gly in place of D Ala. The addition of D-Ala to the culture medium restores production of the D-Ala containing forms of HC-toxin by the toxG mutant. The toxG mutant has only partially reduced virulence. It is concluded that TOXG encodes an alanine racemase whose function is to synthesize D-Ala for incorporation into HC-toxin. PMID- 10671528 TI - Reconciling structure and function in HhaI DNA cytosine-C-5 methyltransferase. AB - Pre-steady state partitioning analysis of the HhaI DNA methyltransferase directly demonstrates the catalytic competence of the enzyme.DNA complex and the lack of catalytic competence of the enzyme.S-adenosyl-L-methionine (AdoMet) complex. The enzyme.AdoMet complex does form, albeit with a 50-fold decrease in affinity compared with the ternary enzyme.AdoMet.DNA complex. These findings reconcile the distinct binding orientations previously observed within the binary enzyme.AdoMet and ternary enzyme. S-adenosyl-L-homocysteine.DNA crystal structures. The affinity of the enzyme for DNA is increased 900-fold in the presence of its cofactor, and the preference for hemimethylated DNA is increased to 12-fold over unmethylated DNA. We suggest that this preference is partially due to the energetic cost of retaining a cavity in place of the 5-methyl moiety in the ternary complex with the unmethylated DNA, as revealed by the corresponding crystal structures. The hemi- and unmethylated substrates alter the fates and lifetimes of discrete enzyme.substrate intermediates during the catalytic cycle. Hemimethylated substrates partition toward product formation versus dissociation significantly more than unmethylated substrates. The mammalian DNA cytosine-C-5 methyltransferase Dnmt1 shows an even more pronounced partitioning toward product formation. PMID- 10671529 TI - Probing the structure of photosystem II with amines and phenylhydrazine. AB - Photosynthetic oxygen evolution is catalyzed at the manganese-containing active site of photosystem II (PSII). Amines are analogs of substrate water and inhibitors of oxygen evolution. Recently, the covalent incorporation of (14)C from [(14)C]methylamine and benzylamine into PSII subunits has been demonstrated (Ouellette, A. J. A., Anderson, L. B., and Barry, B. A. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 2204-2209). To obtain more information concerning these labeling reactions, t-[(14)C]butylamine and phenylhydrazine were employed as probes. Neither compound can be oxidized by a transamination or addition/elimination mechanism, but both can react with activated carbonyl groups, produced as a result of posttranslational modification of amino acid residues, to give amine-derived adducts. (14)C incorporation into the PSII subunits D2/D1 and CP47 was obtained upon treatment of PSII with either t [(14)C]butylamine or [(14)C]phenylhydrazine. For t-butylamine and methylamine, the amount of labeling increased when PSII was treated with denaturing agents. Labeling of CP47, D2, and D1 with methylamine and phenylhydrazine approached a one-to-one stoichiometry, assuming that D2 and D1 each have one binding site. Evidence was obtained suggesting that reductive stabilization and/or access are modulated by PSII light reactions. These results support the proposal that PSII subunits D2, D1, and CP47 contain quinocofactors and that access to these sites is sterically limited. PMID- 10671530 TI - General anesthetic action at an internal protein site involving the S4-S5 cytoplasmic loop of a neuronal K(+) channel. AB - The structural bases of general anesthetic action on a neuronal K(+) channel were investigated using the series of homologous 1-alkanols, electrophysiology, and mutational analysis. Domain swapping between dShaw2 (alkanol-sensitive) and hKv3.4 (alkanol-resistant) and site-directed mutagenesis demonstrated that a 13 amino acid cytoplasmic loop (S4-S5) determines the selective inhibition of native dShaw2 channels by 1-alkanols. The S4-S5 loop may contribute to a receptor for both 1-alkanols and the inactivation particle, because the enhanced 1-alkanol sensitivity of hKv3.4 channels hosting S4-S5 mutations correlates directly with disrupted channel inactivation. Evidence of a discrete protein site was also obtained from the analysis of the relationship between potency and alkyl chain length, which begins to level off after 1-hexanol. Rapid application to the cytoplasmic side of inside-out membrane patches shows that the interaction between dShaw2 channels and 1-alkanols equilibrates in <200 ms. By contrast, the equilibration time is >1000-fold slower when the drug is applied externally to outside-out membrane patches. The data strongly favor a mechanism of inhibition involving a discrete internal site for 1-alkanols in dShaw2 K(+) channels. A new working hypothesis proposes that 1-alkanols lock dShaw2 channels in their closed conformation by a direct interaction at a crevice formed by the S4-S5 loop. PMID- 10671531 TI - In vitro selection of RNA molecules that inhibit the activity of ricin A-chain. AB - The cytotoxin ricin disables translation by depurinating a conserved site in eukaryotic rRNA. In vitro selection has been used to generate RNA ligands (aptamers) specific for the catalytic ricin A-chain (RTA). The anti-RTA aptamers bear no resemblance to the normal RTA substrate, the sarcin-ricin loop (SRL), and were not depurinated by RTA. An initial 80-nucleotide RNA ligand was minimized to a 31-nucleotide aptamer that contained all sequences and structures necessary for interacting with RTA. This minimal RNA formed high affinity complexes with RTA (K(d) = 7.3 nM) which could compete directly with the SRL for binding to RTA. The aptamer inhibited RTA depurination of the SRL and could partially protect translation from RTA inhibition. The IC(50) of the aptamer for RTA in an in vitro translation assay is 100 nM, roughly 3 orders of magnitude lower than a small molecule inhibitor of ricin, pteroic acid, and 2 orders of magnitude lower than the best known RNA inhibitor. The novel anti-RTA aptamers may find application as diagnostic reagents for a potential biological warfare agent and hold promise as scaffolds for the development of strong ricin inhibitors. PMID- 10671532 TI - RNA aptamers that bind to and inhibit the ribosome-inactivating protein, pepocin. AB - Pepocin, isolated from Cucurbita pepo, is a ribosome-inactivating protein (RIP). RIPs site-specifically recognize and depurinate an adenosine at position 4324 in rat 28 S rRNA, rendering the ribosome incapable of interacting with essential elongation factors. Aptamers that target pepocin were isolated from a degenerate RNA pool by in vitro selection. A conserved hairpin motif, quite different from the sequence of the toxin-substrate domain in rat 28 S rRNA, was identified in the aptamer sequences. The aptamers selectively bind to pepocin with dissociation constants between 20 and 30 nM and inhibit the N-glycosidase activity of pepocin on rat liver 28 S rRNA. Competitive binding experiments using aptamer variants suggest that the conserved hairpin region in the anti-pepocin aptamer binds near the catalytic site on pepocin and prevents the interaction of pepocin and 28 S rRNA. Anti-RIP aptamers have potential use in diagnostic systems for the detection of pepocin or could be used as therapy to prevent the action of pepocin in mammalian cells. PMID- 10671533 TI - Benzo[a]pyrene induces the transcription of cyclooxygenase-2 in vascular smooth muscle cells. Evidence for the involvement of extracellular signal-regulated kinase and NF-kappaB. AB - Polycyclic aromatic hydrocarbons, such as benzo[a]pyrene (B[a]P) present in tobacco smoke and tar, have been implicated in the development of atherosclerosis as well as cancer. Increased expression of cyclooxygenase-2 (COX-2) has been detected both in atherosclerotic lesions and in epithelial cancers. To determine whether polycyclic aromatic hydrocarbons might directly affect COX expression in vascular cells, we investigated the effects of B[a]P on COX-2 expression in human and rat arterial smooth muscle cells (SMC). Treatment with B[a]P increased levels of COX-2 protein and mRNA and enhanced prostaglandin synthesis. Nuclear runoff assays and transient transfections revealed increased COX-2 gene transcription after treatment with B[a]P. Experiments were done to define the signaling mechanism by which B[a]P induced COX-2. B[a]P caused a rapid increase in phosphorylation of extracellular signal-regulated kinase (ERK); pharmacologic inhibition of mitogen-activated protein kinase kinase blocked B[a]P-mediated induction of COX-2. Depletion of the intracellular antioxidant, glutathione, with buthionine sulfoximine significantly increased B[a]P-mediated induction of COX-2 while exposure to N-acetylcysteine, a precursor of glutathione, suppressed the induction of COX-2 by B[a]P. Several lines of evidence suggest that the induction of COX-2 by B[a]P is mediated, at least in part, by NF-kappaB. Treatment with B[a]P increased binding of NF-kappaB to DNA. Moreover, B[a]P-mediated stimulation of COX-2 promoter activity was blocked when a construct containing a mutagenized NF-kappaB site was used. Pharmacological inhibitors of NF-kappaB blocked the induction of COX-2 protein and the stimulation of COX-2 promoter activity by B[a]P. Taken together, these data are likely to be important for understanding the atherogenic effects of tobacco smoke. PMID- 10671534 TI - Distinct repair activities of human 7,8-dihydro-8-oxoguanine DNA glycosylase and formamidopyrimidine DNA glycosylase for formamidopyrimidine and 7,8-dihydro-8 oxoguanine. AB - 7,8-dihydro-8-oxoguanine (8-oxoG) and 2,6-diamino-4-hydroxyformamidopyrimidine (Fapy) are major DNA lesions formed by reactive oxygen species and are involved in mutagenic and/or lethal events in cells. Both lesions are repaired by human 7, 8-dihydro-8-oxoguanine DNA glycosylase (hOGG1) and formamidopyrimidine DNA glycosylase (Fpg) in human and Escherichia coli cells, respectively. In the present study, the repair activities of hOGG1 and Fpg were compared using defined oligonucleotides containing 8-oxoG and a methylated analog of Fapy (me-Fapy) at the same site. The k(cat)/K(m) values of hOGG1 for 8-oxoG and me-Fapy were comparable, and this was also the case for Fpg. However, the k(cat)/K(m) values of hOGG1 for both lesions were approximately 80-fold lower than those of Fpg. Analysis of the Schiff base intermediate by NaBH(4) trapping implied that lower substrate affinity and slower hydrolysis of the intermediate for hOGG1 than Fpg accounted for the difference. hOGG1 and Fpg showed distinct preferences of the base opposite 8-oxoG, with the activity differences being 19.8- (hOGG1) and 12 fold (Fpg) between the most and least preferred bases. Surprisingly, such preferences were almost abolished and less than 2-fold for both enzymes when me Fapy was a substrate, suggesting that, unlike 8-oxoG, me-Fapy is not subjected to paired base-dependent repair. The repair efficiency of me-Fapy randomly incorporated in M13 DNA varied at the sequence level, but orders of preferred and unpreferred repair sites were quite different for hOGG1 and Fpg. The distinctive activities of hOGG1 and Fpg including enzymatic parameters (k(cat)/K(m)), paired base, and sequence context effects may originate from the differences in the inherent architecture of the DNA binding domain and catalytic mechanism of the enzymes. PMID- 10671535 TI - Human peroxisomal multifunctional enzyme type 2. Site-directed mutagenesis studies show the importance of two protic residues for 2-enoyl-CoA hydratase 2 activity. AB - Beta-oxidation of acyl-CoAs in mammalian peroxisomes can occur via either multifunctional enzyme type 1 (MFE-1) or type 2 (MFE-2), both of which catalyze the hydration of trans-2-enoyl-CoA and the dehydrogenation of 3-hydroxyacyl-CoA, but with opposite chiral specificity. Amino acid sequence alignment of the 2 enoyl-CoA hydratase 2 domain in human MFE-2 with other MFE-2s reveals conserved protic residues: Tyr-347, Glu-366, Asp-370, His-406, Glu-408, Tyr-410, Asp-490, Tyr-505, Asp-510, His-515, Asp-517, and His-532. To investigate their potential roles in catalysis, each residue was replaced by alanine in site-directed mutagenesis, and the resulting constructs were tested for complementation in a yeast. After additional screening, the wild type and noncomplementing E366A and D510A variants were expressed and characterized. The purified proteins have similar secondary structural elements, with the same subunit composition. The E366A variant had a k(cat)/K(m) value 100 times lower than that of the wild type MFE-2 at pH 5, whereas the D510A variant was inactive. Asp-510 was imbedded in a novel hydratase 2 motif found in the hydratase 2 proteins. The data show that the hydratase 2 reaction catalyzed by MFE-2 requires two protic residues, Glu-366 and Asp-510, suggesting that their catalytic role may be equivalent to that of the two catalytic residues of hydratase 1. PMID- 10671536 TI - Cloning, expression, characterization, and nucleophile identification of family 3, Aspergillus niger beta-glucosidase. AB - The beta-glucosidase from Aspergillus niger (CMI CC 324262) was purified, and an N-terminal sequence and two internal sequences were determined. BglI genomic gene and the cDNA were cloned from a genomic library and by reverse transcriptase polymerase chain reaction, respectively. The cDNA was successfully expressed in Saccharomyces cerevisiae and Pichia pastoris. Sequence analysis revealed that the gene encodes a 92-kDa enzyme that is a member of glycosidase family 3. (1)H-NMR analysis of the reaction catalyzed by this enzyme confirmed that, in common with other family 3 glycosidases, this enzyme hydrolyzes with net retention of anomeric configuration. Accordingly, the enzyme was inactivated by 2-deoxy-2 fluoro beta-glucosyl fluoride, with kinetic parameters of k(i) = 4.5 min(-1), K(I) = 35.4 mM, through the trapping of a covalent glycosyl enzyme intermediate. The catalytic competence of this intermediate was demonstrated by the fact that incubation with linamarin resulted in reactivation, presumably via a transglycosylation mechanism. Peptic digestion of the 2-deoxy-2-fluoroglucosyl enzyme and subsequent analysis of high pressure liquid chromatography eluates by electrospray ionization triple quadrupole mass spectrometry in the neutral loss mode allowed the localization of a 2-deoxy-2-fluoroglucosyl-peptide. Sequence determination of this labeled peptide by tandem mass spectrometry in the daughter ion scan mode permitted the identification of Asp-261 as the catalytic nucleophile within the sequence VMSDW. Asp-261 is fully conserved within this family, consistent with its key role, and aligns with the aspartic acid residue previously identified in the Aspergillus wentii enzyme by labeling with conduritol B epoxide (Bause, E., and Legler, G. (1974) Hoppe-Seyler's Z. Physiol. Chem. 355, 438-442). PMID- 10671537 TI - Mannose trimming targets mutant alpha(2)-plasmin inhibitor for degradation by the proteasome. AB - We have previously characterized the molecular and cellular mechanisms of alpha(2)-plasmin inhibitor (alpha(2)PI) deficiency. The mutant alpha(2)PI-Nara and alpha(2)PI-Okinawa proteins were found to be retained and degraded in cells stably expressing these mutant forms of alpha(2)PI. Degradation of the two mutant alpha(2)PI proteins, mediated by proteasomes, occurred after a lag time of 1.5 h during which glucose trimming took place. The mutant alpha(2)PI proteins were not ubiquitinated. Inhibition of mannosidase activity blocked the degradation of the mutant alpha(2)PI proteins without resulting in any changes in their binding to calnexin. Inhibition of glucose removal completely blocked the interaction between the alpha(2)PI proteins and the molecular chaperone calnexin. Under these conditions, mannose residues were removed from the oligosaccharides even when glucose residues were not processed. With mannose removal, the glucose-untrimmed mutant forms of alpha(2)PI, which failed to bind to calnexin, were degraded by proteasomes. The initiation of mannose trimming was a prerequisite for their degradation. Our findings show that modification of oligosaccharides of the mutant forms of alpha(2)PI determines their recognition by the degradation apparatus and that mannose trimming is important for targeting the mutant alpha(2)PI proteins for the degradation pathway. PMID- 10671538 TI - Cloning and characterization of the alpha(1,3/4) fucosyltransferase of Helicobacter pylori. AB - The gastric pathogen Helicobacter pylori can express the histo blood group antigens, which are on the surface of many human cells. Most H. pylori strains express the type II carbohydrates, Lewis X and Y, whereas a small population express the type I carbohydrates, Lewis A and B. The expression of Lewis A and Lewis X, as in the case of H. pylori strain UA948, requires the addition of fucose in alpha1,4 and alpha1,3 linkages to type I or type II carbohydrate backbones, respectively. This work describes the cloning and characterization of a single H. pylori fucosyltransferase (FucT) enzyme, which has the ability to transfer fucose to both of the aforementioned linkages in a manner similar to the human fucosyltransferase V (Fuc-TV). Two homologous copies of the fucT gene have been identified in each of the genomes sequenced. The characteristic adenosine and cytosine tracts in the amino terminus and repeated regions in the carboxyl terminus are present in the DNA encoding the two UA948fucT genes, but these genes also contain differences when compared with previously identified H. pylori fucTs. The UA948fucTa gene encodes an approximately 52-kDa protein containing 475 amino acids, whereas UA948fucTb does not encode a full-length FucT protein. In vitro, UA948FucTa appears to add fucose with a greater than 5-fold preference for type II chains but still retains significant activity using type I acceptors. The addition of the fucose to the type II carbohydrate acceptors, by UA948FucTa, does not appear to be affected by fucosylation at other sites on the carbohydrate acceptor, but the rate of fucose transfer is affected by terminal fucosylation of type I acceptors. Through mutational analysis we demonstrate that only FucTa is active in this H. pylori isolate and that inactivation of this enzyme eliminates expression of all Lewis antigens. PMID- 10671539 TI - A pattern-recognition protein for beta-1,3-glucan. The binding domain and the cDNA cloning of beta-1,3-glucan recognition protein from the silkworm, Bombyx mori. AB - The beta-1,3-glucan recognition protein (betaGRP) has strong specific affinity for beta-1,3-glucan, a component of the fungal cell wall. Its interaction with beta-1,3-glucan initiates the activation of the prophenoloxidase cascade, which is an important defense system in invertebrates of many species. We cloned the cDNA of the betaGRP of the silkworm Bombyx mori. The betaGRP mRNA transcript was constitutively expressed in the hemocytes, fat body, and epithelial cells of the naive silkworm. At the same time, a bacterial or yeast challenge was indicated to intensify the transcription. Comparison of the deduced amino acid sequence with known sequences revealed that the betaGRP contained a region (Thr(264) to Pro(386)) displaying significant similarity to the catalytic regions of bacterial beta-1,3-glucanases and much higher similarity to the glucanase-like regions of Gram-negative bacteria-binding proteins found in the silkworm B. mori and the mosquito Anopheles gambiae. The region (Thr(264) to Pro(386)) of the betaGRP, however, was demonstrated not to have appreciable affinity for beta-1,3-glucan. A recombinant peptide corresponding to an N-terminal region (Tyr(1) to Ala(102)) of the betaGRP bound strongly to beta-1,3-glucan. These results indicate that the binding domain of the betaGRP for beta-1,3-glucan is located in the N-terminal region. Glucanases and the current pattern-recognition proteins that contain a glucanase-like region seem to have a common origin in their molecular evolution. PMID- 10671540 TI - The regulatory beta subunit of protein kinase CK2 mediates formation of tetrameric CK2 complexes. AB - Protein kinase CK2 is a tetrameric enzyme composed of two catalytic (alpha and/or alpha') subunits and two regulatory (beta) subunits. Because CK2beta is synthesized in excess of CK2alpha, we hypothesized that formation of CK2beta homodimers precedes the incorporation of the catalytic subunits of CK2 into complexes. To test this hypothesis, we cotransfected cells with two epitope tagged variants of CK2beta. The results of these cotransfection studies demonstrate that interactions between two CK2beta subunits take place in the absence of CK2alpha. Together with results from previous biosynthetic labeling studies, these results suggest that formation of CK2beta homodimers occurs before incorporation of catalytic subunits of CK2 into CK2 complexes. We also cotransfected Cos-7 cells with a deletion fragment of CK2beta (i.e. Myc-beta1 166) together with full-length hemagglutinin (HA)-tagged CK2beta and/or CK2alpha'. Although complexes between Myc-beta1-166 and HA-beta were readily detected, we obtained no evidence of direct interactions between Myc-beta1-166 and HA-CK2alpha'. These results suggest that residues within the N-terminal 166 amino acids of CK2beta are sufficient for interactions between CK2beta subunits, whereas the C-terminal domain of CK2beta is required for complex formation with the catalytic subunits of CK2. Finally, we observed that expression of full length HA-beta promotes phosphorylation of Myc-beta1-166 by HA-CK2alpha'. PMID- 10671541 TI - Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes. AB - Adipocyte lipolysis was compared with hormone-sensitive lipase (HSL)/perilipin subcellular distribution and perilipin phosphorylation using Western blot analysis. Under basal conditions, HSL resided predominantly in the cytosol and unphosphorylated perilipin upon the lipid droplet. Upon lipolytic stimulation of adipocytes isolated from young rats with the beta-adrenergic agonist, isoproterenol, HSL translocated from the cytosol to the lipid droplet, but there was no movement of perilipin from the droplet to the cytosol; however, perilipin phosphorylation was observed. By contrast, upon lipolytic stimulation and perilipin phosphorylation in cells from more mature rats, there was no HSL translocation but a significant movement of perilipin away from the lipid droplet. Adipocytes from younger rats had markedly greater rates of lipolysis than those from the older rats. Thus high rates of lipolysis require translocation of HSL to the lipid droplet and translocation of HSL and perilipin can occur independently of each other. A loss of the ability to translocate HSL to the lipid droplet probably contributes to the diminished lipolytic response to catecholamines with age. PMID- 10671542 TI - Fatty acid and lipoic acid biosynthesis in higher plant mitochondria. AB - Fatty acid and lipoic acid biosynthesis were investigated in plant mitochondria. Although the mitochondria lack acetyl-CoA carboxylase, our experiments reveal that they contain the enzymatic equipment necessary to transform malonate into the two main building units for fatty acid synthesis: malonyl- and acetyl-acyl carrier protein (ACP). We demonstrated, by a new method based on a complementary use of high performance liquid chromatography and mass spectrometry, that the soluble mitochondrial fatty-acid synthase produces mainly three predominant acyl ACPs as follows: octanoyl(C8)-, hexadecanoyl(C16)-, and octadecanoyl(C18)-ACP. Octanoate production is of primary interest since it has been postulated long ago to be a precursor of lipoic acid. By using a recombinant H apoprotein mutant as a potential acceptor for newly synthesized lipoic acid, we were able to detect limited amounts of lipoylated H protein in the presence of malonate, several sulfur donors, and cofactors. Finally, we present a scheme outlining the new biochemical pathway of fatty acid and lipoic acid synthesis in plant mitochondria. PMID- 10671543 TI - Estrogen stimulates heat shock protein 90 binding to endothelial nitric oxide synthase in human vascular endothelial cells. Effects on calcium sensitivity and NO release. AB - Estradiol (E(2)) causes endothelium-dependent vasodilation, mediated, in part, by enhanced nitric oxide (NO) release. We have previously shown that E(2)-induced activation of endothelial nitric oxide synthase (eNOS) reduces its calcium dependence. This pathway of eNOS activation is unique to a limited number of stimuli, including shear stress, the response to which is herbimycin-inhibitable. Consistent with this, herbimycin and geldanamycin pretreatment of human umbilical vein endothelial cells (HUVEC) abrogated E(2)-stimulated NO release and cGMP production, respectively. These benzoquinone ansamycins are potent inhibitors of Hsp90 function, which has recently been shown to play a role in stimulus dependent eNOS activation. As in response to shear, E(2) induced an Hsp90-eNOS association, peaking at 30 min and completely inhibited by the conventional estrogen receptor antagonist ICI 182,780. These findings suggest that Hsp90 plays an important role in the rapid, estrogen receptor-mediated modulation of eNOS activation by estrogen. PMID- 10671544 TI - Apoptotic cleavage of scaffold attachment factor A (SAF-A) by caspase-3 occurs at a noncanonical cleavage site. AB - Members of the caspase family of cysteine proteases play essential roles in the disintegration of cellular architecture during apoptosis. Caspases have been grouped into subfamilies according to their preferred cleavage sites, with the "apoptotic executioner" caspase-3 as the prototype of DEXD-dependent proteases. We show here that caspase-3 is more tolerant to variations of the cleavage site than previously anticipated and present an example of a noncanonical recognition site that is efficiently cleaved by caspase-3 in vitro and in vivo. The new cleavage site was identified in human scaffold attachment factor A, one of the major scaffold attachment region DNA-binding proteins of human cells thought to be involved in nuclear architecture by fastening chromatin loops to a proteinaceous nuclear skeleton, the so-called nuclear matrix or scaffold. Using an amino-terminal recombinant construct of scaffold attachment factor A and recombinant caspase-3, we have mapped the cleavage site by matrix-assisted laser desorption ionization/time of flight mass spectrometry and Edman sequencing. We find that cleavage occurs after Asp-100 in a sequence context (SALD) that does not conform to the hitherto accepted DEXD consensus sequence of caspase-3. A point mutation, D100A, abrogates cleavage by recombinant caspase-3 in vitro and during apoptosis in vivo, confirming SALD as a novel caspase-3 cleavage site. PMID- 10671545 TI - Parathyroid hormone regulation of the rat collagenase-3 promoter by protein kinase A-dependent transactivation of core binding factor alpha1. AB - Previously we showed that the activator protein-1 site and the runt domain binding site in the collagenase-3 promoter act cooperatively in response to parathyroid hormone (PTH) in the rat osteoblastic osteosarcoma cell line, UMR 106 01. Our results of the expression pattern of core binding factor alpha1 (Cbfa1), which binds to the runt domain site, indicated that there is no change in the levels of Cbfa1 protein or RNA under either control conditions or after PTH treatment. The importance of posttranslational modification of Cbfa1 in the signaling pathway for PTH-induced collagenase-3 promoter activity was analyzed. PTH stimulation of collagenase-3 promoter activity was completely abrogated by protein kinase A (PKA) inhibition. To determine the role of PKA activity with respect to Cbfa1 activation (in addition to its known activity of phosphorylating cAMP-response element-binding protein to enhance c-fos promoter activity), we utilized the heterologous Gal4 transcription system. PTH stimulated the transactivation of activation domain-3 in Cbfa1 through the PKA site. In vitro phosphorylation studies indicated that the PKA site in the wild type activation domain-3 is a substrate for phosphorylation by PKA. Thus, we demonstrate that PTH induces a PKA-dependent transactivation of Cbfa1, and this transactivation is required for collagenase-3 promoter activity in UMR cells. PMID- 10671546 TI - Distinct central amphipathic alpha-helices in apolipoprotein A-I contribute to the in vivo maturation of high density lipoprotein by either activating lecithin cholesterol acyltransferase or binding lipids. AB - Recombinant adenoviruses with cDNAs for human apolipoprotein A-I (wild type (wt) apoA-I) and three mutants, referred to as Delta4-5A-I, Delta5-6A-I, and Delta6-7A I, that have deletions removing regions coding for amino acids 100-143, 122-165, and 144-186, respectively, were created to study structure/function relationships of apoA-I in vivo. All mutants were expressed at lower concentrations than wt apoA-I in plasma of fasting apoA-I-deficient mice. The Delta5-6A-I mutant was found primarily in the lipid-poor high density lipoprotein (HDL) pool and at lower concentrations than Delta4-5A-I and Delta6-7A-I that formed more buoyant HDL(2/3) particles. At an elevated adenovirus dose and earlier blood sampling from fed mice, both Delta5-6A-I and Delta6-7A-I increased HDL-free cholesterol and phospholipid but not cholesteryl ester. In contrast, wt apoA-I and Delta4-5A I produced significant increases in HDL cholesteryl ester. Further analysis showed that Delta6-7A-I and native apoA-I could bind similar amounts of phospholipid and cholesterol that were reduced slightly for Delta5-6A-I and greatly for Delta4-5A-I. We conclude from these findings that amino acids (aa) 100-143, specifically helix 4 (aa 100-121), contributes to the maturation of HDL through a role in lipid binding and that the downstream sequence (aa 144-186) centered around helix 6 (aa 144-165) is responsible for the activation of lecithin-cholesterol acyltransferase. PMID- 10671547 TI - Removal of a putative inhibitory element reduces the calcium-dependent calmodulin activation of neuronal nitric-oxide synthase. AB - Neuronal nitric-oxide synthase (NOS) and endothelial NOS are constitutive NOS isoforms that are activated by binding calmodulin in response to elevated intracellular calcium. In contrast, the inducible NOS isoform binds calmodulin at low basal levels of calcium in resting cells. Primary sequence comparisons show that each constitutive NOS isozyme contains a polypeptide segment within its reductase domain, which is absent in the inducible NOS enzyme. To study a possible link between the presence of these additional polypeptide segments in constitutive NOS enzymes and their calcium-dependent calmodulin activation, three deletion mutants were created. The putative inhibitory insert was removed from the FMN binding regions of the neuronal NOS holoenzyme and from two truncated neuronal NOS reductase enzymes in which the calmodulin binding region was either included or deleted. All three mutant enzymes showed reduced incorporation of FMN and required reconstitution with exogenous FMN for activity. The combined removal of both the calmodulin binding domain and the putative inhibitory insert did not result in a calmodulin-independent neuronal NOS reductase. Thus, although the putative inhibitory element has an effect on the calcium-dependent calmodulin activation of neuronal NOS, it does not have the properties of the typical autoinhibitory domain found in calmodulin-activated enzymes. PMID- 10671548 TI - Class- and splice variant-specific association of CD98 with integrin beta cytoplasmic domains. AB - CD98 is a type II transmembrane protein involved in neutral and basic amino acid transport and in cell fusion events. CD98 was implicated in the function of integrin adhesion receptors by its capacity to reverse suppression of integrin activation by isolated integrin beta(1A) domains. Here we report that CD98 associates with integrin beta cytoplasmic domains with a unique integrin class and splice variant specificity. In particular, CD98 interacted with the ubiquitous beta(1A) but not the muscle-specific splice variant, beta(1D), or leukocyte-specific beta(7) cytoplasmic domains. The ability of CD98 to associate with integrin cytoplasmic domains correlated with its capacity to reverse suppression of integrin activation. The association of CD98 with integrin beta(1A) cytoplasmic domains may regulate the function and localization of these membrane proteins. PMID- 10671549 TI - Cu,Zn-superoxide dismutase-dependent apoptosis induced by nitric oxide in neuronal cells. AB - Nitric oxide (NO) challenge to human neuroblastoma cells (SH-SY5Y) ultimately results in apoptosis. Tumor suppressor protein p53 and cell cycle inhibitor p21 accumulate as an early sign of S-nitrosoglutathione-mediated toxicity. Cytochrome c release from mitochondria and caspase 3 activation also occurred. Cells transfected with either wild type (WT) or mutant (G93A) Cu, Zn-superoxide dismutase (Cu,Zn-SOD) produced comparable amounts of nitrite/nitrate but showed different degree of apoptosis. G93A cells were the most affected and WT cells the most protected; however, Cu, Zn-SOD content of these two cell lines was 2-fold the SH-SY5Y cells under both resting and treated conditions. We linked decreased susceptibility of the WT cells to higher and more stable Bcl-2 and decreased reactive oxygen species. Conversely, we linked G93A susceptibility to increased reactive oxygen species production since simultaneous administration of S nitrosoglutathione and copper chelators protects from apoptosis. Furthermore, G93A cells showed a significant decrease of Bcl-2 expression and, as target of NO derived radicals, showed lower cytochrome c oxidase activity. These results demonstrate that resistance to NO-mediated apoptosis is strictly related to the level and integrity of Cu,Zn-SOD and that the balance between reactive nitrogen and reactive oxygen species regulates neuroblastoma apoptosis. PMID- 10671550 TI - The differentially conserved residues of carbamoyl-phosphate synthetase. AB - Carbamoyl-phosphate synthetase (CPS) from Escherichia coli is a heterodimeric protein. The larger of the two subunits (M(r) approximately 118,000) contains a pair of homologous domains of approximately 400 residues each that are approximately 40% identical in amino acid sequence. The carboxy phosphate (residues 1-400) and carbamoyl phosphate domains (residues 553-933) also contain approximately 79 differentially conserved residues. These are residues that are conserved throughout the bacterial evolution of CPS in one of these homologous domains but not the other. The role of these differentially conserved residues in the structural and catalytic properties of CPS was addressed by swapping segments of these residues from one domain to the other. Nine of these chimeric mutant enzymes were constructed, expressed, purified, and characterized. A majority of the mutants were unable to synthesize any carbamoyl phosphate and the rest were severely crippled. True tandem repeat chimeric proteins were constructed by the complete substitution of one homologous domain sequence for the other. Neither of the two possible chimeric proteins was structurally stable. These results have been interpreted to demonstrate that the two homologous domains in the large subunit of CPS are functionally and structurally nonequivalent. This nonequivalence is a direct result of the specific functions each of these domains must perform during the overall synthesis of carbamoyl phosphate in the wild type enzyme and the specific structural alterations imposed by the differentially conserved residues. PMID- 10671551 TI - The thioredoxin system of Helicobacter pylori. AB - This paper describes the purification of thioredoxin reductase (TR) and the characterization, purification, and cloning of thioredoxin (Trx) from Helicobacter pylori. Purification, amino acid sequence analysis, and molecular cloning of the gene encoding thioredoxin revealed that it is a 12-kDa protein which possesses the conserved redox active motif CGPC. The gene encoding Trx was amplified by polymerase chain reaction and inserted into a pET expression vector and used to transform Escherichia coli. Trx was overexpressed by induction with isopropyl-1-thio-beta-D-galactopyranoside as a decahistidine fusion protein and was recovered from the cytoplasm as a soluble and active protein. The redox activity of this protein was characterized using several mammalian proteins of different architecture but all containing disulfide bonds. H. pylori thioredoxin efficiently reduced insulin, human immunoglobulins (IgG/IgA/sIgA), and soluble mucin. Subcellular fractionation analysis of H. pylori revealed that thioredoxin was associated largely with the cytoplasm and inner membrane fractions of the cell in addition to being recovered in the phosphate-buffered saline-soluble fraction of freshly harvested cells. H. pylori TR was purified to homogeneity by chromatography on DEAE-52, Cibacron blue 3GA, and 2',5'-ADP-agarose. Gel filtration revealed that the native TR had a molecular mass of 70 kDa which represented a homodimer composed of two 35-kDa subunits, as determined by SDS polyacrylamide gel electrophoresis. H. pylori TR (NADPH-dependent) efficiently catalyzed the reduction of 5,5'-dithiobis(nitrobenzoic acid) in the presence of either native or recombinant H. pylori Trx. H. pylori Trx behaved also as a stress response element as broth grown bacteria secreted Trx in response to chemical, biological, and environmental stresses. These observations suggest that Trx may conceivably assist H. pylori in the process of colonization by inducing focal disruption of the oligomeric structure of mucin while rendering host antibody inactive through catalytic reduction. PMID- 10671552 TI - Casein kinase II phosphorylation of E-cadherin increases E-cadherin/beta-catenin interaction and strengthens cell-cell adhesion. AB - Beta-catenin, a member of the Armadillo repeat protein family, binds directly to the cytoplasmic domain of E-cadherin, linking it via alpha-catenin to the actin cytoskeleton. A 30-amino acid region within the cytoplasmic domain of E-cadherin, conserved among all classical cadherins, has been shown to be essential for beta catenin binding. This region harbors several putative casein kinase II (CKII) and glycogen synthase kinase-3beta (GSK-3beta) phosphorylation sites and is highly phosphorylated. Here we report that in vitro this region is indeed phosphorylated by CKII and GSK-3beta, which results in an increased binding of beta-catenin to E cadherin. Additionally, in mouse NIH3T3 fibroblasts expression of E-cadherin with mutations in putative CKII sites resulted in reduced cell-cell contacts. Thus, phosphorylation of the E-cadherin cytoplasmic domain by CKII and GSK-3beta appears to modulate the affinity between beta-catenin and E-cadherin, ultimately modifying the strength of cell-cell adhesion. PMID- 10671553 TI - Utilization of distinct signaling pathways by receptors for vascular endothelial cell growth factor and other mitogens in the induction of endothelial cell proliferation. AB - This study was initiated to identify signaling proteins used by the receptors for vascular endothelial cell growth factor KDR/Flk1, and Flt1. Two-hybrid cloning and immunoprecipitation from human umbilical vein endothelial cells (HUVEC) showed that KDR binds to and promotes the tyrosine phosphorylation of phospholipase Cgamma (PLCgamma). Neither placental growth factor, which activates Flt1, epidermal growth factor (EGF), or fibroblast growth factor (FGF) induced tyrosine phosphorylation of PLCgamma, indicating that KDR is uniquely important to PLCgamma activation in HUVEC. By signaling through KDR, VEGF promoted the tyrosine phosphorylation of focal adhesion kinase, induced activation of Akt, protein kinase Cepsilon (PKCepsilon), mitogen-activated protein kinase (MAPK), and promoted thymidine incorporation into DNA. VEGF activates PLCgamma, PKCepsilon, and phosphatidylinositol 3-kinase independently of one another. MEK, PLCgamma, and to a lesser extent PKC, are in the pathway through which KDR activates MAPK. PLCgamma or PKC inhibitors did not affect FGF- or EGF-mediated MAPK activation. MAPK/ERK kinase inhibition diminished VEGF-, FGF-, and EGF promoted thymidine incorporation into DNA. However, blockade of PKC diminished thymidine incorporation into DNA induced by VEGF but not FGF or EGF. Signaling through KDR/Flk1 activates signaling pathways not utilized by other mitogens to induce proliferation of HUVEC. PMID- 10671554 TI - Cloning and expression of glycolipid transfer protein from bovine and porcine brain. AB - Glycolipid transfer protein (GLTP) is a small (23-24 kDa), basic protein (pI congruent with 9.0) that accelerates the intermembrane transfer of various glycolipids. Here, we report the first cloning of cDNAs that encode the bovine and porcine GLTPs. The cDNA open reading frame for bovine GLTP was constructed by bridge-overlapping extension polymerase chain reaction (PCR) after obtaining partial coding cDNA clones by hot start, seminested, and rapid amplification of cDNA ends-PCR. The cDNA open reading frame for porcine GLTP was constructed by reverse transcriptase-PCR. The encoded amino acid sequences in the full-length bovine and porcine cDNAs were identical, consisting of 209 amino acid residues, and were nearly the same as the published sequence determined by Edman degradation. The cDNA encoded one additional amino acid at the N terminus (methionine), arginine at positions 10 and 200 instead of lysine, and threonine at position 65 instead of alanine. Expression of GLTP-cDNA in Escherichia coli using pGEX-6P-1 vector resulted in glutathione S-transferase (GST)-GLTP fusion protein. Regulation of growth and induction conditions led to approximately 50% of expressed fusion protein being soluble and active. Proteolytic cleavage of GST GLTP fusion protein (bound to GST-Sepharose) and affinity purification resulted in fully active GLTP. Northern blot analyses of bovine tissues showed a single transcript of approximately 2.2 kilobases and the following hierarchy of mRNA levels: cerebrum > kidney > spleen congruent with lung congruent with cerebellum > liver > heart muscle. Reverse transcriptase-PCR analyses of mRNA levels supported the Northern blot results. PMID- 10671555 TI - E5 oncoprotein mutants activate phosphoinositide 3-kinase independently of platelet-derived growth factor receptor activation. AB - The E5 oncoprotein of bovine papillomavirus type 1 is a Golgi-resident, 44-amino acid polypeptide that can transform fibroblast cell lines by activating endogenous platelet-derived growth factor receptor beta (PDGF-R). However, the recent discovery of E5 mutants that exhibit strong transforming activity but minimal PDGF-R tyrosine phosphorylation indicates that E5 can potentially use additional signal transduction pathway(s) to transform cells. We now show that two classes of E5 mutants, despite poorly activating the PDGF-R, induce tyrosine phosphorylation and activation of phosphoinositide 3-kinase (PI 3-K) and that this activation is resistant to a selective inhibitor of PDGF-R kinase activity, tyrphostin AG1296. Consistent with this independence from PDGF-R signaling, the E5 mutants fail to induce significant cell proliferation in the absence of PDGF, unlike wild-type E5 or the sis oncoprotein. Despite differences in growth factor requirements, however, both wild-type E5 and mutant E5 cell lines form colonies in agarose. Interestingly, activation of PI 3-K occurs without concomitant activation of the ras-dependent mitogen-activated protein kinase pathway. The known ability of constitutively activated PI 3-K to induce anchorage-independent cell proliferation suggests a mechanism by which the mutant E5 proteins transform cells. PMID- 10671556 TI - Catalytic sites for 3' and 5' incision of Escherichia coli nucleotide excision repair are both located in UvrC. AB - Nucleotide excision repair in Escherichia coli is a multistep process in which DNA damage is removed by incision of the DNA on both sides of the damage, followed by removal of the oligonucleotide containing the lesion. The two incision reactions take place in a complex of damaged DNA with UvrB and UvrC. It has been shown (Lin, J. -J., and Sancar, A. (1992) J. Biol. Chem. 267, 17688 17692) that the catalytic site for incision on the 5' side of the damage is located in the UvrC protein. Here we show that the catalytic site for incision on the 3' side is in this protein as well, because substitution R42A abolishes 3' incision, whereas formation of the UvrBC-DNA complex and the 5' incision reaction are unaffected. Arg(42) is part of a region that is homologous to the catalytic domain of the homing endonuclease I-TevI. We propose that the UvrC protein consists of two functional parts, with the N-terminal half for the 3' incision reaction and the C-terminal half containing all the determinants for the 5' incision reaction. PMID- 10671557 TI - The role of disulfide-linked dimerization in interleukin-3 receptor signaling and biological activity. AB - Cysteine residues 86 and 91 of the beta subunit of the human interleukin (hIL)-3 receptor (hbetac) participate in disulfide-linked receptor subunit heterodimerization. This linkage is essential for receptor tyrosine phosphorylation, since the Cys-86 --> Ala (Mc4) and Cys-91 --> Ala (Mc5) mutations abolished both events. Here, we used these mutants to examine whether disulfide-linked receptor dimerization affects the biological and biochemical activities of the IL-3 receptor. Murine T cells expressing hIL-3Ralpha and Mc4 or Mc5 did not proliferate in hIL-3, whereas cells expressing wild-type hbetac exhibited rapid proliferation. However, a small subpopulation of cells expressing each mutant could be selected for growth in IL-3, and these proliferated similarly to cells expressing wild-type hbetac, despite failing to undergo IL-3 stimulated hbetac tyrosine phosphorylation. The Mc4 and Mc5 mutations substantially reduced, but did not abrogate, IL-3-mediated anti-apoptotic activity in the unselected populations. Moreover, the mutations abolished IL-3 induced JAK2, STAT, and AKT activation in the unselected cells, whereas activation of these molecules in IL-3-selected cells was normal. In contrast, Mc4 and Mc5 showed a limited effect on activation of Erk1 and -2 in unselected cells. These data suggest that whereas disulfide-mediated cross-linking and hbetac tyrosine phosphorylation are normally important for receptor activation, alternative mechanisms can bypass these requirements. PMID- 10671558 TI - Direct cleavage by the calcium-activated protease calpain can lead to inactivation of caspases. AB - Caspases, a unique family of cysteine proteases involved in cytokine activation and in the execution of apoptosis can be sub-grouped according to the length of their prodomain. Long prodomain caspases such as caspase-8 and caspase-9 are believed to act mainly as upstream caspases to cleave downstream short prodomain caspases such as caspases-3 and -7. We report here the identification of caspases as direct substrates of calcium-activated proteases, calpains. Calpains cleave caspase-7 at sites distinct from those of the upstream caspases, generating proteolytically inactive fragments. Caspase-8 and caspase-9 can also be directly cleaved by calpains. Two calpain cleavage sites in caspase-9 have been identified by N-terminal sequencing of the cleaved products. Cleavage of caspase-9 by calpain generates truncated caspase-9 that is unable to activate caspase-3 in cell lysates. Furthermore, direct cleavage of caspase-9 by calpain blocks dATP and cytochrome-c induced caspase-3 activation. Therefore our results suggest that calpains may act as negative regulators of caspase processing and apoptosis by effectively inactivating upstream caspases. PMID- 10671559 TI - Cholesterol depletion of enterocytes. Effect on the Golgi complex and apical membrane trafficking. AB - Intestinal brush border enzymes, including aminopeptidase N and sucrase isomaltase, are associated with "rafts" (membrane microdomains rich in cholesterol and sphingoglycolipids). To assess the functional role of rafts in the present work, we studied the effect of cholesterol depletion on apical membrane trafficking in enterocytes. Cultured mucosal explants of pig small intestine were treated for 2 h with the cholesterol sequestering agent methyl beta-cyclodextrin and lovastatin, an inhibitor of hydroxymethylglutaryl-coenzyme A reductase. The treatment reduced the cholesterol content >50%. Morphologically, the Golgi complex/trans-Golgi network was partially transformed into numerous 100 200 nm vesicles. By immunogold electron microscopy, aminopeptidase N was localized in these Golgi-derived vesicles as well as at the basolateral cell surface, indicating a partial missorting. Biochemically, the rates of the Golgi associated complex glycosylation and association with rafts of newly synthesized aminopeptidase N were reduced, and less of the enzyme had reached the brush border membrane after 2 h of labeling. In contrast, the basolateral Na(+)/K(+) ATPase was neither missorted nor raft-associated. Our results implicate the Golgi complex/trans-Golgi network in raft formation and suggest a close relationship between this event and apical membrane trafficking. PMID- 10671560 TI - Functional association between SLAP-130 and SLP-76 in Jurkat T cells. AB - T cell antigen receptor (TCR) engagement results in protein-tyrosine kinase activation which initiates signaling cascades leading to induction of the interleukin-2 gene. Previous studies identified two substrates of the TCR-induced protein-tyrosine kinases, SH2 domain-containing leukocyte specific protein of 76 kDa (SLP-76) and SLP-76-associated phosphoprotein of 130 kDa (SLAP-130). While SLP-76 appears to couple the TCR with downstream signals, SLAP-130 may play a negative regulatory role in T cell activation. In this study, we demonstrate that consistent with its ability to abrogate the SLP-76 augmentation of TCR-induced activation of the NFAT/AP1 region of the interleukin-2 promoter, overexpression of SLAP-130 also interferes with the rescue of signaling in SLP-76-deficient Jurkat cells in co-transfection experiments. The effect of SLAP-130 on SLP-76 function is specific for regulating TCR-induced ERK activation, but not phospholipase Cgamma 1 phosphorylation. By generating both deletion and point mutants of SLAP-130, we identified tyrosine 559 as critical for the interaction between SLP-76 and SLAP-130. We show that mutation of this residue in context of full-length SLAP-130 diminishes the ability of SLAP-130 to abrogate SLP-76 function. These data suggest that the SLAP-130/SLP-76 association is important for the negative regulatory role that SLAP-130 appears to play in T cell signaling. PMID- 10671561 TI - Structure and activity associated with multiple forms of Schizosaccharomyces pombe DNA polymerase delta. AB - DNA polymerase delta (Pol delta) isolated from Schizosaccharomyces pombe (sp) consists of at least four subunits, Pol3, Cdc1, Cdc27, and Cdm1. We have reconstituted the four-subunit complex by simultaneously expressing these polypeptides in baculovirus-infected insect cells. The properties of the purified cloned spPol delta were identical to the native spPol delta isolated from S. pombe cells. In addition, we also isolated a three-subunit complex containing Pol3, Cdc1, and Cdm1. Both three- and four-subunit complexes required replication factor C and proliferating cell nuclear antigen for DNA replication. However, in the presence of low levels of polymerase complexes, the three-subunit complex was less efficient than the four-subunit complex in supporting DNA replication. The inefficient synthesis of DNA by the three-subunit complex can be remedied by the addition of Cdc27, the subunit missing in the three-subunit complex. Gel filtration analysis demonstrated that the three-subunit complex is a monomer of the heterotrimer (Pol3, Cdc1, and Cdm1) and that the four-subunit complex is a dimer of the heterotetramer (Pol3, Cdc1, Cdc27, and Cdm1), similar to the structure of native spPol delta. We have further shown that Cdc1 and Cdc27 interact to form a heterodimeric complex. Gel filtration studies indicate that the structure of this complex is dimeric. These observations suggest that the Cdc27 subunit may play an important role contributing to the dimerization of Pol delta. PMID- 10671562 TI - Effects of jasplakinolide on the kinetics of actin polymerization. An explanation for certain in vivo observations. AB - Jasplakinolide paradoxically stabilizes actin filaments in vitro, but in vivo it can disrupt actin filaments and induce polymerization of monomeric actin into amorphous masses. A detailed analysis of the effects of jasplakinolide on the kinetics of actin polymerization suggests a resolution to this paradox. Jasplakinolide markedly enhances the rate of actin filament nucleation. This increase corresponds to a change in the size of actin oligomer capable of nucleating filament growth from four to approximately three subunits, which is mechanistically consistent with the localization of the jasplakinolide-binding site at an interface of three actin subunits. Because jasplakinolide both decreases the amount of sequestered actin (by lowering the critical concentration of actin) and augments nucleation, the enhancement of polymerization by jasplakinolide is amplified in the presence of actin-monomer sequestering proteins such as thymosin beta(4). Overall, the kinetic parameters in vitro define the mechanism by which jasplakinolide induces polymerization of monomeric actin in vivo. Expected consequences of jasplakinolide function are consistent with the experimental observations and include de novo nucleation resulting in disordered polymeric actin and in insufficient monomeric actin to allow for remodeling of stress fibers. PMID- 10671563 TI - Two heme-binding domains of heme-regulated eukaryotic initiation factor-2alpha kinase. N terminus and kinase insertion. AB - In heme deficiency, protein synthesis in reticulocytes is inhibited by activation of heme-regulated alpha-subunit of eukaryotic initiation factor-2alpha (eIF 2alpha) kinase (HRI). Previous studies indicate that HRI contains two distinct heme-binding sites per HRI monomer. To study the role of the N terminus in the heme regulation of HRI, two N-terminally truncated mutants, Met2 and Met3 (deletion of the first 103 and 130 amino acids, respectively), were prepared. Met2 and Met3 underwent autophosphorylation and phosphorylated eIF-2alpha with a specific activity of approximately 50% of that of the wild type HRI. These mutants were significantly less sensitive to heme regulation both in vivo and in vitro. In addition, the heme contents of purified Met2 and Met3 HRI were less than 5% of that of the wild type HRI. These results indicated that the N terminus was important but was not the only domain involved in the heme-binding and heme regulation of HRI. Heme binding of the individual HRI domains showed that both N terminus and kinase insertion were able to bind hemin, whereas the C terminus and the catalytic domains were not. Thus, both the N terminus and the kinase insertion, which are unique to HRI, are involved in the heme binding and the heme regulation of HRI. PMID- 10671564 TI - Endothelial nitric-oxide synthase (type III) is activated and becomes calcium independent upon phosphorylation by cyclic nucleotide-dependent protein kinases. AB - Endothelial nitric-oxide synthase (NOS-III) is defined as being strictly dependent on Ca(2+)/calmodulin (CaM) for activity, although NO release from endothelial cells has been reported to also occur at intracellular free Ca(2+) levels that are substimulatory for the purified enzyme. We demonstrate here that NOS-III, but neither NOS-I nor -II, is rapidly and strongly activated and phosphorylated on both Ser and Thr in the presence of cGMP-dependent protein kinase II (cGK II) and the catalytic subunit of cAMP-dependent protein kinase (cAK) in vitro. Phosphopeptide analysis by mass spectrometry identified Ser(1177), as well as Ser(633) which is situated in a recently defined CaM autoinhibitory domain within the flavin-binding region of human NOS-III. Phosphoamino acid analysis identified a putative phosphorylation site at Thr(495) in the CaM-binding domain. Importantly, both cAK and cGK phosphorylation of NOS III in vitro caused a highly reproducible partial (10-20%) NOS-III activation which was independent of Ca(2+)/CaM, and as much as a 4-fold increase in V(max) in the presence of Ca(2+)/CaM. cAK stimulation in intact endothelial cells also increased both Ca(2+/)CaM-independent and -dependent activation of NOS-III. These data collectively provide new evidence for cAK and cGK stimulation of both Ca(2+)/CaM-independent and -dependent NOS-III activity, and suggest possible cross-talk between the NO and prostaglandin I(2) pathways and a positive feedback mechanism for NO/cGMP signaling. PMID- 10671565 TI - Alternative splicing of GAD67 results in the synthesis of a third form of glutamic-acid decarboxylase in human islets and other non-neural tissues. AB - Two forms of glutamic-acid decarboxylase (GAD) have been identified in mammalian tissues: a 65-kDa form (GAD65) and a 67-kDa form (GAD67). Alternate splicing produces one or two smaller variants of GAD67 in the brain of embryonic mice and rats. Additionally, a short, heretofore unidentified transcript homologous to GAD67 has been detected in human testis RNA. Because GAD, the enzyme responsible for gamma-aminobutyric acid production and a key autoantigen in type I diabetes, has unclear function in non-neural tissue, it is important to understand its pattern of expression. Unlike GAD65, GAD67 is not produced in human pancreatic islets. Here, we describe a novel splice variant of GAD67 that is produced in human islets, testis, adrenal cortex, and perhaps other endocrine tissues, but not in brain. This transcript directs the synthesis of a protein without GAD enzymatic activity: GAD25. A unique peptide sequence at the carboxyl terminus of GAD25 is highly conserved between mice, rats, and humans. We conclude that humans produce a third form of GAD in non-neural tissues and that human islets, although they do not synthesize full-length GAD67, do express this shortened variant. PMID- 10671566 TI - LeSBT1, a subtilase from tomato plants. Overexpression in insect cells, purification, and characterization. AB - The cDNA of a tomato subtilase designated LeSBT1 was cloned from a tomato flower cDNA library. The deduced amino acid sequence indicated for LeSBT1 the structure of a prepro-protein targeted to the secretory pathway by virtue of an amino terminal signal peptide. LeSBT1 was expressed in the baculovirus/insect cell system and a processed 73-kDa form of LeSBT1, lacking both signal peptide and prodomain, was purified to homogeneity from culture supernatants. This 73-kDa LeSBT1, however, lacked proteolytic activity. Zymogen activation to yield 68-kDa LeSBT1 required the additional processing of an amino-terminal autoinhibitory peptide in a strictly pH-dependent manner. Mature 68-kDa LeSBT1 showed highest activity at acidic pH consistent with its presumed localization in the apoplast of the plant cell. In comparison to other plant subtilases, LeSBT1 exhibited a narrower substrate specificity in that it cleaves only polypeptide substrates preferentially but not exclusively carboxyl-terminal of glutamine residues. The possible involvement of LeSBT1 in selective proprotein processing is discussed with reference to the related mammalian proprotein convertases. PMID- 10671567 TI - Glucose regulation of mouse S(14) gene expression in hepatocytes. Involvement of a novel transcription factor complex. AB - Transcription of genes encoding enzymes required for lipogenesis is induced in hepatocytes in response to elevated glucose metabolism. We have previously mapped the carbohydrate-response elements (ChoREs) of the rat liver-type pyruvate kinase (L-PK) and S(14) genes and found them to share significant sequence similarity. However, progress in unraveling this signaling pathway has been hampered due to the difficulty in identifying the key factor(s) that bind to these ChoREs. To gain further insight into the nature of the carbohydrate-responsive transcription factor, the glucose regulatory sequences from the mouse S(14) gene were examined in primary hepatocytes. Three elements were found to be essential for supporting the glucose response: a thyroid hormone-response element between -1522 and -1494, an accessory factor site between -1421 and -1392, and the ChoRE between -1450 and -1425. Of these, only the accessory factor site was conserved between the rat and mouse S(14) genes. Investigation of the ChoRE sequence indicated that two half E box motifs are critical for the response to glucose. Electrophoretic mobility shift assays revealed a complex formed between the mouse S(14) ChoRE and liver nuclear proteins. This complex was also formed by ChoREs from the rat S(14) and L PK genes but not by mutants of these sites that are inactive in supporting the glucose response. These results suggest the presence of a novel transcription factor complex that mediates the glucose-regulated transcription of S(14) and L PK genes. PMID- 10671568 TI - Requirement for protein-tyrosine phosphatase SHP-2 in insulin-induced activation of c-Jun NH(2)-terminal kinase. AB - Mitogen-activated protein kinases, including extracellular signal-regulated kinases and c-Jun NH(2)-terminal kinases (JNKs), are activated by insulin. Although the mechanism by which the insulin receptor activates extracellular signal-regulated kinases is relatively well defined, the pathway that leads to JNK activation is poorly understood. Overexpression of a catalytically inactive mutant (SHP-2C/S) of the protein-tyrosine phosphatase SHP-2 in Rat-1 fibroblasts that also express human insulin receptors has now revealed that activation of JNKs by insulin and epidermal growth factor, but not that by anisomycin or sorbitol, requires SHP-2. A dominant negative mutant (RasN17) of Ha-Ras blocked insulin-induced JNK activation, whereas a dominant negative mutant (RacN17) of Rac1 or a specific inhibitor (LY294002) of phosphoinositide 3-kinase did not, indicating a role for Ras, but not for Rac or phosphoinositide 3-kinase, in this effect. SHP-2C/S markedly inhibited Ras activation in response to insulin without affecting insulin-induced tyrosine phosphorylation of cellular substrates or the dissociation of the Crk-p130(Cas) complex. In contrast, SHP-2C/S did not inhibit activation of JNKs induced by a constitutively active mutant (RasV12) of Ha-Ras. Furthermore, expression of myristoylated SOS, which functions as a potent activator of Ras, induced JNK activation even when SHP-2 was inactivated. These results suggest that SHP-2 contributes to JNK activation in response to insulin by positively regulating the Ras signaling pathway at the same level as, or upstream from, SOS. PMID- 10671569 TI - Identification of a novel sterol-independent regulatory element in the human low density lipoprotein receptor promoter. AB - The cytokine oncostatin M (OM) activates human low density lipoprotein receptor (LDLR) gene transcription through a sterol-independent mechanism. Previous studies conducted in our laboratory have narrowed the OM-responsive element to promoter region -52 to +13, which contains the repeat 3 and two TATA-like sequences. We now identify LDLR promoter region -17 to -1 as a sterol-independent regulatory element (SIRE) that is critically involved in OM-, transcription factor CCAAT/enhancer-binding protein (C/EBP)-, and second messenger cAMP mediated activation of LDLR transcription. The SIRE sequence overlaps the previously described TATA-like element and consists of an active C/EBP-binding site (-17 to -9) and a functional cAMP-responsive element (CRE) (-8 to -1). We demonstrate that (a) mutations within either the C/EBP or CRE site have no impact on basal or cholesterol-mediated repression of LDLR transcription, but they completely abolish OM-mediated activation of LDLR transcription; (b) replacing the repeat 3 sequence that contains the Sp1-binding site with a yeast transcription factor GAL4-binding site in the LDLR promoter construct does not affect OM inducibility, thereby demonstrating that OM induction is mediated through the SIRE sequence in conjunction with a strong activator bound to the repeat 3 sequence; (c) electrophoretic mobility shift and supershift assays confirm the specific binding of transcription factors C/EBP and cAMP-responsive element-binding protein to the SIRE; (d) cotransfection of a human C/EBPbeta expression vector (pEF-NFIL6) with the LDLR promoter construct pLDLR234 increases LDLR promoter activity; and (e) OM and dibutyryl cAMP synergistically activate LDLR transcription through this regulatory element. This study identifies, for the first time, a cis-acting regulatory element in the LDLR promoter that is responsible for sterol-independent regulation of LDLR transcription. PMID- 10671570 TI - R-Ras contains a proline-rich site that binds to SH3 domains and is required for integrin activation by R-Ras. AB - R-Ras contains a proline-rich motif that resembles SH3 domain-binding sites but that has escaped notice previously. We show here that this site in R-Ras is capable of binding SH3 domains and that the SH3 domain binding may be important for R-Ras function. A fusion protein containing the SH3 domains of the adaptor protein Nck interacted strongly with the R-Ras proline-rich sequence and with the intact protein. The binding was independent of whether R-Ras was in its GDP or GTP form. The Nck binding, which was mediated by the second of the three SH3 domains of Nck, was obliterated by mutations in the proline-rich sequence of R Ras. The interaction of Nck with R-Ras could also be shown in yeast two-hybrid assays and by co-immunoprecipitation in human cells transfected with Nck and R Ras. Previous results have shown that the expression of a constitutively active R Ras mutant, R-Ras(38V), converts mouse 32D monocytic cells into highly adherent cells. Introducing the proline mutations into R-Ras(38V) suppressed the effect of R-Ras on 32D cell adhesion while not affecting GTP binding. These results reveal an unexpected regulatory pathway that controls R-Ras through an SH3 domain interaction. This pathway appears to be important for the ability of R-Ras to control cell adhesion. PMID- 10671571 TI - Platelet-derived growth factor rapidly increases activity and cell surface expression of the EAAC1 subtype of glutamate transporter through activation of phosphatidylinositol 3-kinase. AB - Na(+)-dependent glutamate transporters are the primary mechanism for removal of excitatory amino acids (EAAs) from the extracellular space of the central nervous system and influence both physiologic and pathologic effects of these compounds. Recent evidence suggests that the activity and cell surface expression of a neuronal subtype of glutamate transporter, EAAC1, are rapidly increased by direct activation of protein kinase C and are decreased by wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-K). We hypothesized that this regulation could be analogous to insulin-induced stimulation of the GLUT4 subtype of glucose transporter, which is dependent upon activation of PI3-K. Using C6 glioma, a cell line that endogenously and selectively expresses EAAC1, we report that platelet derived growth factor (PDGF) increased Na(+)-dependent L-[(3)H]-glutamate transport activity within 30 min. This effect of PDGF was not due to a change in total cellular EAAC1 immunoreactivity but was instead correlated with an increase cell surface expression of EAAC1, as measured using a membrane impermeant biotinylation reagent combined with Western blotting. A decrease in nonbiotinylated intracellular EAAC1 was also observed. These studies suggest that PDGF causes a redistribution of EAAC1 from an intracellular compartment to the cell surface. These effects of PDGF were accompanied by a 35-fold increase in PI3 K activity and were blocked by the PI3-K inhibitors, wortmannin and LY 294002, but not by an inhibitor of protein kinase C. Other growth factors, including insulin, nerve growth factor, and epidermal growth factor had no effect on glutamate transport nor did they increase PI3-K activity. These studies suggest that, as is observed for insulin-mediated translocation of GLUT4, EAAC1 cell surface expression can be rapidly increased by PDGF through activation of PI3-K. It is possible that this PDGF-mediated increase in EAAC1 activity may contribute to the previously demonstrated neuroprotective effects of PDGF. PMID- 10671572 TI - Essential role of human leukocyte antigen-encoded proteasome subunits in NF kappaB activation and prevention of tumor necrosis factor-alpha-induced apoptosis. AB - The multisubunit proteasome complex is the principal mediator of nonlysosomal protein degradation. The proteasome subunit varies minimally between cells with the exception of LMP2, LMP7, and LMP10 subunits in rodent and human cells. LMP2 and LMP7 subunits are encoded by the human lymphocyte antigen region, and they optimize proteolytic mediated antigen presentation. The proteasome is also important for the function of transcription factor nuclear factor-kappaB (NF kappaB). It is required for NF-kappaB subunits p50 and p52 generation and catalyzes degradation of phosphorylated IkappaBalpha. These proteasome-mediated reactions have now been shown to be defective in T2 cells, a human lymphocyte cell line that lacks both LMP2 and LMP7. Although T2 cells contain normal expression of p100 and p105, the abundance of p50 and p52 was greatly reduced. Tumor necrosis factor-alpha (TNF-alpha) induced normal phosphorylation of IkappaBalpha but failed to induce degradation of phosphorylated IkappaBalpha. Both DNA binding assays and luciferase assays revealed that TNF-alpha-induced NF kappaB activation is defective in T2 cells. Unlike parental cells, T2 cells were susceptible to TNF-alpha-induced apoptosis. These data indicate human leukocyte antigen-linked proteasome subunits are essential for NF-kappaB activation and protection of cells from TNF-alpha-induced apoptosis. PMID- 10671573 TI - Thoracic radiology: the past 50 years. AB - The radiology of 50 years ago was a primitive science compared with the radiology of today. Hospital departments were small and radiologists few in number. Night call was uncommon. Examinations consisted primarily of radiographs of the chest, bones, and gastrointestinal tract, although some early neuroradiologic studies were performed. Chest fluoroscopy was common. Film processing was done manually, often with poor results. Radiographic examinations of the chest were likewise unsophisticated by today's standards. Chest radiographs were made with low kilovoltage, calcium tungstate phosphors and relatively large focal spots. There were no image intensifiers, nuclear medicine studies, ultrasonography, computed tomography, or magnetic resonance studies. How far we have come! PMID- 10671574 TI - 2000 RSNA leadership PMID- 10671575 TI - An endangered art: teaching. PMID- 10671576 TI - Public policy: the case for lobbying in radiology. PMID- 10671577 TI - MR angiography in the evaluation of atherosclerotic peripheral vascular disease. AB - Magnetic resonance (MR) angiography of lower extremity occlusive vascular disease has evolved into a feasible diagnostic imaging option. The previous emphasis on time-of-flight techniques was associated with lengthy acquisition times and artifactual signal losses. Those limitations presented an obstacle to widespread clinical implementation. However, the emergence of rapid imaging sequences combined with gadolinium chelate enhancement offers time-efficient alternatives that can yield a truer representation of the vascular anatomic structure. The technology is now poised to serve as a routine screening study, provided that radiologists understand all factors needed to generate clinically relevant MR angiograms. This article is intended to provide a useful resource directed toward achieving that understanding. PMID- 10671578 TI - The dog leg sign. PMID- 10671579 TI - Symptomatic vertebral hemangiomas: treatment by means of direct intralesional injection of ethanol. AB - PURPOSE: To describe the technique and results of injecting ethanol directly into symptomatic vertebral hemangiomas. MATERIALS AND METHODS: Eleven patients with paraplegia (n = 6) or radiculopathy (n = 5) due to vertebral hemangioma were treated by means of injecting ethanol (5-50 mL) directly into the lesion with computed tomographic (CT) guidance. CT angiograms were essential prior to treatment to identify functional vascular spaces of the hemangioma and direct needle placement. RESULTS: All hemangiomas were obliterated completely at follow up angiography and gadolinium-enhanced magnetic resonance imaging. Five of six patients with paraplegia recovered completely: One who was treated recently was walking with assistance. Four of five patients with radiculopathy improved. No immediate complications were associated with ethanol injection. The two patients who received the largest volumes of ethanol, 42 and 50 mL, developed pathologic fractures of the involved vertebrae 4 and 16 weeks after treatment. CONCLUSION: Direct injection of ethanol into symptomatic vertebral hemangioma is an effective and safe treatment, provided the dose is less than 15 mL. PMID- 10671580 TI - Transarterial chemoembolization for hepatocellular carcinoma: volumetric and morphologic CT criteria for assessment of prognosis and therapeutic success results from a liver transplantation center. AB - PURPOSE: To evaluate the prognostic value of volumetric computed tomography (CT) for therapy control in patients treated with repeated transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Eighty-five patients with histologically proved HCC underwent 182 TACE procedures with 50 mg/m(2) doxorubicin hydrochloride, 50 mg/m(2) cisplatin, 10 mL/m(2) iodized oil, and amilomer microspheres. The volumes of liver and tumor were measured with a region-of-interest CT technique. Iodized oil retention was estimated with CT performed 24-48 hours after treatment. RESULTS: Tumor volume expressed as a percentage of liver volume was less than 5% in 26, less than 15% in 33, and 15% or greater in 26 patients. The overall 1-year survival rate was 57.6% (mean, 534 days; median, 428 days). There was a statistically significant prolongation of survival when the tumor volume was less than 200 mL (P <.02) and less than 5% of the liver volume (P <.01). Complete (>/=75%) and good (50%-74%) iodized oil retention raised the median survival significantly (P <.001 and P <.07, respectively). Significantly reduced survival correlated with diffuse tumor growth pattern (P <.05) and presence of more than nine lesions (P <.03). CONCLUSION: TACE resulted in significant prolongation of survival in patients with tumor volumes of less than 200 mL, tumor-to-liver volume ratios of less than 5%, and iodized oil retention greater than or equal to 75%. PMID- 10671581 TI - Percutaneous incision of stenotic uroenteric anastomoses with a cutting balloon catheter: long-term results. AB - PURPOSE: To describe the technique and results of incision of strictures in anastomotic urinary diversions with a commercially available cutting balloon catheter. MATERIALS AND METHODS: Thirty-seven stenoses were treated in 32 patients. Most (28 [88%]) of the patients had undergone surgery for bladder cancer 17.7 months +/- 17.4 (SD) (range, 3-72 months) before incision. Thirteen patients had undergone ileal conduit diversion, and nineteen had undergone enterocystoplasty. All stenoses were shorter than 3 cm. The presence of adjacent ileal loops and/or iliac vessels was assessed with computed tomography before incision. The cutting wire was oriented anteriorly or anterolaterally, and the balloon was inflated with diluted contrast material during the incision. A Kaplan Meier survival curve was constructed to illustrate the success rates over time. RESULTS: No major complications occurred. Twelve (32%) stenoses recurred in nine patients 15 months +/- 10 (range, 6-36 months) after stent removal; the failure rate was 53% (eight of 15 stenoses) for ileal conduits and 18% (four of 22 stenoses) for enterocystoplasties. Late failure (>12 months) was observed in four patients. The patency of the other 25 stenoses (23 patients) was checked 25 months +/- 11 after stent removal (range, 5-43 months). The actuarial patency rate was 77% at 1 year, 68% at 2 years, and 62% at 3 years. CONCLUSION: Cutting balloon incision is a safe and simple alternative to surgery, particularly when the urinary diversion is enterocystoplasty. PMID- 10671582 TI - Improved uniformity of aortic enhancement with customized contrast medium injection protocols at CT angiography. AB - PURPOSE: To compare the uniformity of aortoiliac opacification obtained from uniphasic contrast medium injections versus individualized biphasic injections at computed tomographic (CT) angiography. MATERIALS AND METHODS: Thirty-two patients with an abdominal aortic aneurysm underwent CT angiography. In 16 patients (group 1), 120 mL of contrast material was administered at a flow rate of 4 mL/sec. In the other 16 patients (group 2), biphasic injection protocols were computed by using mathematic deconvolution of each patient's time-attenuation response to a standardized test injection. Attenuation uniformity was quantified as the "plateau deviation" of enhancement values, which were calculated as the SD of the time-contiguous attenuation values observed during the 30-second scanning period. RESULTS: Group 2 patients received between 77 and 165 mL (mean, 115 mL) of contrast medium. Initial flow rates ranged from 4.1 to 10.0 mL/sec (mean, 6.8 mL/sec) for the first 4-6 seconds; continuing flow rates ranged from 2.0 to 4.8 mL/sec (mean, 3.1 mL/sec) for the remaining 24-26 seconds. The plateau deviation was significantly smaller in group 2 patients (19 HU) versus group 1 patients (38 HU, P <.001). CONCLUSION: At CT angiography, tailored biphasic injections led to more uniform aortoiliac enhancement, compared with standard uniphasic injections of contrast medium. PMID- 10671583 TI - Renal arterial stenosis: prospective comparison of color Doppler US and breath hold, three-dimensional, dynamic, gadolinium-enhanced MR angiography. AB - PURPOSE: To compare color Doppler ultrasonography (US) with fast, breath-hold, three-dimensional, gadolinium-enhanced magnetic resonance (MR) angiography in detecting renal arterial stenosis. MATERIALS AND METHODS: Forty-five patients with clinical suspicion of renovascular disease were prospectively examined with intra- and extrarenal color Doppler US and breath-hold, gadolinium-enhanced MR angiography. Digital subtraction arteriography (DSA) was the standard of reference in all patients for the number of renal arteries and degree of stenosis. RESULTS: DSA depicted 103 arteries and 52 stenoses. Color Doppler US was nondiagnostic in two examinations. Significantly more of 13 accessory renal arteries were detected with MR angiography (n = 12) than with color Doppler US (n = 3; P <.05). For assessing all stenoses, the sensitivity and accuracy were 94% and 91%, respectively, for MR angiography and 71% and 76%, respectively, for US (P <.05). The sensitivity was higher for MR angiography (100%) than for US (79%; P <.05) in diagnosing stenoses with at least 50% narrowing. The specificity, accuracy, and negative predictive value in diagnosing stenoses of at least 50% narrowing were 93%, 95%, and 100% for MR angiography and 93%, 89%, and 90% for US. CONCLUSION: Breath-hold, gadolinium-enhanced MR angiography is superior to color Doppler US in accessory renal artery detection. Although the specificity of MR angiography is similar to that of color Doppler US, MR angiography has a better sensitivity and negative predictive value in depicting renal arterial stenoses. PMID- 10671584 TI - Vascularity of hepatocellular carcinoma: assessment with contrast-enhanced second harmonic versus conventional power Doppler US. AB - PURPOSE: To compare contrast material-enhanced harmonic power Doppler ultrasonography (US) with conventional power Doppler US in depicting the vascularity of hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Twenty patients with nodular HCCs (2.6-13.2 cm in diameter; mean diameter, 4.8 cm) were prospectively examined with both conventional and harmonic power Doppler US. US was performed with a 2-4-MHz curved linear-array transducer according to a standard examination protocol (1,000-Hz pulse repetition frequency, medium wall filter, and power gain of 55%-84% for conventional power Doppler US; 700-Hz pulse repetition frequency, low wall filter, and power gain of 95%-98% for harmonic power Doppler US). Serial, dynamic scans were obtained before intravenous injection of the contrast agent (SH U 508A) and at 30, 60, 90, 120, 180, 240, and 300 seconds after injection with both techniques. RESULTS: The number of intratumoral power Doppler US signals was similar with both techniques at 30-90 seconds after contrast agent injection; however, after 90 seconds, conventional power Doppler US depicted significantly more signals than did harmonic power Doppler US. Harmonic power Doppler US was superior to conventional power Doppler US in terms of power Doppler artifacts such as "blooming" or motion-related artifacts. CONCLUSION: Although the effective enhancement duration is relatively short compared with that for conventional power Doppler US, contrast-enhanced harmonic power Doppler US can be effective in evaluating the vascularity of HCCs because of the advantage of fewer power Doppler artifacts. PMID- 10671585 TI - Variability of Doppler US measurements along the common carotid artery: effects on estimates of internal carotid arterial stenosis in patients with angiographically proved disease. AB - PURPOSE: To determine the effect of variability of common carotid arterial (CCA) velocities on velocity ratios used to assess internal carotid arterial (ICA) stenosis. MATERIALS AND METHODS: Doppler ultrasonographic (US) velocity measurements were obtained at three levels in the CCA and in the carotid bulb and ICA in all patients referred for carotid US between September 1996 and October 1997. Only ICAs (n = 98, in 57 patients) without ipsilateral CCA disease at angiography were analyzed. The range of CCA peak systolic velocities (PSVs) and end diastolic velocities (EDVs) and velocity ratios were calculated for each CCA measurement. For each ICA/CCA velocity ratio, receiver operating characteristic analysis was performed. RESULTS: CCA PSV and EDV ranges averaged 23.1 cm/sec +/- 15.7 (SD) and 5.1 cm/sec +/- 3.6, respectively. For a given side, the difference averaged 1.0 +/- 1.3 for PSV ratios and 2.7 +/- 6.9 for EDV ratios, depending on where CCA measurements were taken. By using a threshold of 60% stenosis as indication for endarterectomy, variability in CCA velocities could have altered recommendations in 16 (28%) of 57 patients. Receiver operating characteristic analysis showed that ratios made by using the three CCA velocities or their mean were not significantly different. CONCLUSION: Variability in velocity measurements along the course of the CCA in patients with ICA disease can be substantial and can result in inaccuracies in assessment of carotid stenosis. PMID- 10671586 TI - Parathyroid glands: combination of (99m)Tc MIBI scintigraphy and US for demonstration of parathyroid glands and nodules. AB - PURPOSE: To determine the appropriate choice of imaging techniques for localization of nodular lesions of parathyroid glands. MATERIALS AND METHODS: First, computed tomographic (CT), magnetic resonance (MR), ultrasonographic (US), and technetium 99m methoxyisobutyl-isonitrile (MIBI) scintigraphic images in 49 patients with primary hyperparathyroidism were retrospectively evaluated. A single-blind, prospective study that included 16 patients with primary hyperparathyroidism was then conducted. MR, US, scintigraphic, and color Doppler US images of the neck were obtained and analyzed. RESULTS: In the retrospective study, CT, MR imaging, and US had low sensitivity (13%, 17%, and 27%, respectively) and specificity (39%, 65%, and 65%, respectively). Scintigraphy had 57% sensitivity and 85% specificity. In the prospective study, the use of latest generation MR and US equipment and the participation of experienced operators led to improved sensitivity and specificity for these techniques. The combination of US and scintigraphy resulted in improved sensitivity (96%), specificity (83%), and positive and negative predictive values (88% and 94%, respectively), relative to the results obtained with either method alone. Doppler US was of little help in the setting of small glands. CONCLUSIONS: The combination of (99m)Tc MIBI scintigraphy and US performed by well-trained operators with up-to-date instruments appeared to be the best diagnostic tool for the preoperative diagnosis of parathyroid disease. PMID- 10671587 TI - Hippocampal structures: anteroposterior N-acetylaspartate differences in patients with epilepsy and control subjects as shown with proton MR spectroscopic imaging. AB - PURPOSE: To determine the distribution of proton metabolites along the long axis of the hippocampus. MATERIALS AND METHODS: Proton magnetic resonance (MR) spectroscopic imaging measurements were performed in the hippocampi of 14 control subjects and nine patients with unilateral mesial temporal lobe epilepsy. RESULTS: Control subjects showed significantly lower ratios of N-acetylaspartate (NAA) to choline-containing compounds (Ch) and creatine plus phosphocreatine (CR) (NAA/[Cr + Ch]) in the anterior as compared with the posterior part of the hippocampus. Furthermore, a similar anteroposterior (AP) difference in NAA/(Cr + Ch) values was found in both ipsilateral and contralateral hippocampi of patients. In the patients compared with the control subjects, ipsilateral NAA/(Cr + Ch) levels were reduced in every part of hippocampal tissue with an average reduction of 17%, and contralateral NAA/(Cr + Ch) was reduced by about 10%. In the patients compared with the control subjects, the proportional reduction in ipsilateral NAA/(Cr + Ch) was greatest in voxels from anterior hippocampal regions. CONCLUSION: AP differences could be a result of fewer neurons in the anterior compared with the posterior hippocampus or of the increasing thickness of the hippocampus from posterior to anterior, which leads to different contributions from adjacent tissue. Measurements of T2 showed that T2 differences are probably not responsible for these changes. PMID- 10671588 TI - Central nervous system sarcoidosis: follow-up at MR imaging during steroid therapy. AB - PURPOSE: To document the changes observed at sequential magnetic resonance (MR) imaging of sarcoidosis lesions of the central nervous system (CNS) during treatment with corticosteroids. MATERIALS AND METHODS: The abnormalities detected in 24 patients (mean follow-up, 36 months) were compared before and after therapeutic periods (n = 75) that were divided into attack (high-dose), upkeep (decreased-dose), and minimal (low-dose) periods. Parenchymal lesions were classified as type 1 (enhanced with gadolinium), type 2 (demyelinating), or type 3 (lacunar) and were assessed as regressing, stable, or progressing. RESULTS: Seven of the 24 patients had several types of lesions. Isolated type 3 lesions (six patients) were the only lesions not associated with neurologic deficit. Type 1 lesions (13 patients) regressed in 22 of 22 attack periods and progressed in nine of 27 upkeep and minimal periods. MR imaging depicted relapses in patients with multifocal CNS involvement or long-standing CNS impairment or in those who had previously received steroid therapy. Type 2 (seven patients) and type 3 (13 patients) lesions remained stable in 68 of 68 therapeutic periods. Type 1 lesions appeared in three patients with type 2 and type 3 lesions during two upkeep and three minimal periods. Findings at follow-up MR imaging contributed to the reintroduction of high-dose corticosteroid therapy in eight patients. CONCLUSION: MR imaging can be used to differentiate between reversible and irreversible lesions in CNS sarcoidosis. MR imaging can be a useful tool for adjusting treatment to prevent irreversible CNS damage. PMID- 10671589 TI - Hematologic toxic reaction to radiation therapy adjuvant to autologous peripheral blood stem cell transplantation for recurrent or refractory Hodgkin disease. AB - PURPOSE: To evaluate the hematologic toxic reaction to external-beam radiation therapy after high-dose chemotherapy with peripheral blood stem cell (PBSC) support in patients with Hodgkin disease. MATERIALS AND METHODS: A retrospective study of 30 cases of Hodgkin disease in patients who underwent high-dose carmustine, etoposide, and cyclophosphamide chemotherapy with PBSC support was performed. Thirteen patients underwent radiation therapy (28.8-39.0 Gy) a median of 45 days after PBSC repeat infusion. RESULTS: Radiation therapy was delivered as planned, without interruption, in all patients. Five patients developed thrombocytopenia (one with grade 1 thrombocytopenia; two, grade 2; and two, grade 3) and included three with progressive disease prior to radiation therapy and two with a history of prior irradiation. None developed a bleeding complication or required transfusion support. Five patients who underwent irradiation had thrombocytopenia (three with grade 1 and two with grade 2) 100 days after PBSC repeat infusion, compared with three patients (two with grade 1 and one with grade 3) who did not undergo posttransplantation irradiation. At the most recent follow-up, no patient without evidence of disease had a platelet count of less than 100 x 10(9)/L. CONCLUSION: External-beam radiation therapy was well tolerated in the posttransplantation setting in patients with Hodgkin disease. Thrombocytopenia was common but was not related to clinical complications. PMID- 10671590 TI - Multiple pulmonary nodules in AIDS: usefulness of CT in distinguishing among potential causes. AB - PURPOSE: To determine whether the computed tomographic (CT) appearances of multiple pulmonary nodules in patients with acquired immunodeficiency syndrome (AIDS) can help differentiate the potential infectious and neoplastic causes. MATERIALS AND METHODS: The thoracic CT scans obtained in 60 patients with AIDS and multiple pulmonary nodules were reviewed retrospectively by two thoracic radiologists who were blinded to clinical and pathologic data. The scans were evaluated for nodule size, distribution, and morphologic characteristics. CT findings were correlated with final diagnoses. RESULTS: Thirty-six (84%) of 43 patients with opportunistic infection had a predominance of nodules smaller than 1 cm in diameter, whereas 14 (82%) of 17 patients with a neoplasm had a predominance of nodules larger than 1 cm (P <.001). Of the 43 patients with opportunistic infection, 28 (65%) had a centrilobular distribution of nodules; only one (6%) of 17 patients with a neoplasm had this distribution (P <.001). Seven (88%) of eight patients with a peribronchovascular distribution had Kaposi sarcoma (P <.001). CONCLUSION: In patients with AIDS who have multiple pulmonary nodules at CT, nodule size and distribution are useful in the differentiation of potential causes. Nodules smaller than 1 cm, especially those with a centrilobular distribution, are typically infectious. Nodules larger than 1 cm are often neoplastic. A peribronchovascular distribution is suggestive of Kaposi sarcoma. PMID- 10671591 TI - Segmental and subsegmental pulmonary arteries: evaluation with electron-beam versus spiral CT. AB - PURPOSE: To compare contrast agent-enhanced spiral and electron-beam computed tomography (CT) for the analysis of segmental and subsegmental pulmonary arteries. MATERIALS AND METHODS: CT angiography of the pulmonary arteries was performed in 56 patients to rule out pulmonary embolism. Electron-beam CT was performed in 28 patients. The other 28 patients underwent spiral CT with comparable scanning protocols. The depiction of segmental and subsegmental arteries was analyzed by three independent readers. The contrast enhancement in the main pulmonary artery was measured in each patient. RESULTS: Analysis was performed in 1,120 segmental and 2, 240 subsegmental arteries. One segmental (RA7, P =.010) and two subsegmental (LA7b, P =.029; RA6a+b, P =.038) arteries in paracardiac and basal segments of the lung were depicted significantly better with electron-beam CT. There was no statistically significant difference between electron-beam and spiral CT in the total number of analyzable peripheral arteries depicted. The mean contrast enhancement in the main pulmonary artery was 362 HU in electron-beam CT studies versus 248 HU in spiral CT studies. CONCLUSION: Detailed visualization of peripheral pulmonary arteries is well within the scope of advanced CT techniques. Electron-beam CT has minor advantages in analyzing paracardiac arteries, probably because of reduction of motion artifacts and higher contrast enhancement. Further studies are needed to establish whether electron-beam CT allows a more confident diagnosis of emboli in these vessels. PMID- 10671592 TI - Pulmonary lymphangioleiomyomatosis: correlation of ventilation-perfusion scintigraphy, chest radiography, and CT with pulmonary function tests. AB - PURPOSE: To determine the findings on ventilation-perfusion (V-P) scintigrams, computed tomographic (CT) scans, and chest radiographs and correlate them with pulmonary function test results in patients with lymphangioleiomyomatosis. MATERIALS AND METHODS: V-P scintigraphy, chest radiography, conventional and thin section CT, and pulmonary function tests were performed in 39 patients. The images were graded on a scale of 0 (normal) to 3 (severely abnormal). RESULTS: Imaging abnormalities were found on 92% of ventilation scintigrams, 92% of perfusion scintigrams, 79% of chest radiographs, 100% of CT scans, and 100% of thin-section CT scans. On ventilation scintigrams, 28 (72%) patients demonstrated a speckling pattern. On CT scans, all patients had pulmonary cysts. Univariate analysis showed that extent of disease on chest radiographs and CT scans, cyst size, V-P abnormalities, and degree of speckling were inversely correlated with forced expiratory volume in one second (FEV(1)), diffusing capacity of lung for carbon monoxide, and the ratio of FEV(1) to forced vital capacity (FVC) (P <.01) but not with FVC and total lung capacity. Larger cyst size correlated with extent of disease at CT, but not significantly (P =.056). CONCLUSION: Scintigraphic and radiologic abnormalities are seen in a majority of patients with lymphangioleiomyomatosis. On ventilation scintigrams, a frequently seen speckling pattern may be related to accumulation of radionuclide in pulmonary cysts-a hallmark of the disease at CT. Findings with each imaging modality correlate with certain pulmonary functions. PMID- 10671593 TI - Coronary arterial stent patency: assessment with electron-beam CT. AB - PURPOSE: To evaluate electron-beam computed tomography (CT) for stent localization and noninvasive assessment of stent patency in patients with coronary arterial stents and coronary bypass stents. MATERIALS AND METHODS: CT in the single-section volume mode was performed in 202 patients with 321 coronary arterial stents in 221 vessels to localize the stents. Patency was evaluated in the multisection flow mode with an intravenous bolus injection of contrast material. All electron-beam CT images were reviewed by an observer who had no knowledge of the coronary angiographic results. Electron-beam CT findings were then compared with coronary angiographic findings. RESULTS: The stents could be visualized and related to the coronary arterial segments in 216 of 221 vessels with electron-beam CT. Of the 221 vessels, 207 were correctly evaluated with electron-beam CT. Compared with coronary angiography, electron-beam CT permitted the detection of 18 of 23 high-grade stenoses (sensitivity, 78%) and correctly depicted the absence of high-grade stenoses in 189 of 193 vessels with stents (specificity, 98%). Altogether, 18 stenoses were detected correctly at electron beam CT; the interpretation was false-positive in four vessels (positive predictive value, 82% [18/22 vessels]) and false-negative in five (negative predictive value, 97% [189/194 vessels]). CONCLUSION: Electron-beam CT may be helpful in localizing intracoronary stents and assessing stent patency noninvasively to delay the intervals between catheterizations in an increasing number of patients. PMID- 10671594 TI - Three-dimensional systolic strain patterns in the normal human left ventricle: characterization with tagged MR imaging. AB - PURPOSE: To present a database of systolic three-dimensional (3D) strain evolution throughout the normal left ventricle (LV) in humans. MATERIALS AND METHODS: In 31 healthy volunteers, magnetic resonance (MR) tissue tagging and breath-hold MR imaging were used to generate and then detect the motion of transient fiducial markers (ie, tags) in the heart every 32 msec. Strain and motion were calculated from a 3D displacement field that was fit to the tag data. Special indexes of contraction and thickening that were based on multiple strain components also were evaluated. RESULTS: The temporal evolution of local strains was linear during the first half of systole. The peak shortening and thickening strain components were typically greatest in the anterolateral wall, increased toward the apex, and increased toward the endocardium. Shears and displacements were more spatially variable. The two specialized indexes of contraction and thickening had higher measurement precision and tighter normal ranges than did the traditional strain components. CONCLUSION: In this study, the authors noninvasively characterized the normal systolic ranges of 3D displacement and strain evolution throughout the human LV. Comparison against this multidimensional database may permit sensitive detection of systolic LV dysfunction. PMID- 10671595 TI - Arterial switch procedure for D-transposition of the great arteries: quantitative midterm evaluation of hemodynamic changes with cine MR imaging and phase-shift velocity mapping-initial experience. AB - PURPOSE: To evaluate cine magnetic resonance (MR) imaging and phase-shift velocity mapping for assessment of the hemodynamic relevance of stenotic segments or specific hemodynamic changes in the great vessels after an arterial switch procedure for correction of D-transposition of the great arteries. MATERIALS AND METHODS: Twenty consecutive patients (age range, 2-17 years) with an acoustic window that was insufficient for Doppler transthoracic echocardiography were included in the study. Flow and diameter measurements of the pulmonary arterial trunk and its primary branches were performed with phase-shift velocity mapping and cine MR imaging. RESULTS: There were good correlations between pressure gradients in the pulmonary arteries estimated with MR imaging and those measured with Doppler echocardiography (r = 0.83, n = 15) and cardiac catheterization (r = 0.90, n = 13). Cine MR imaging revealed that the diameters of the right and left pulmonary arteries decreased with the expansion of the aorta during systole, which increased the peak velocity. This temporary stenosis was more severe in the right than in the left pulmonary artery and was accompanied by a significantly (P <.05) lower volume flow in the right artery. CONCLUSION: The anatomic situation after arterial switch repair tended to produce temporary stenoses in the primary pulmonary arterial branches, with significant changes in hemodynamics. These changes may affect the long-term outcome and go undetected with other imaging modalities. PMID- 10671596 TI - Pancreatoblastoma: imaging findings in 10 patients and review of the literature. AB - PURPOSE: To describe the features of pancreatoblastoma at magnetic resonance (MR) imaging, computed tomography (CT), and ultrasonography (US). MATERIALS AND METHODS: Imaging and surgical findings in 10 patients (age range, 2-20 years; mean age, 6.8 years) with pathologically proved pancreatoblastoma were reviewed for tumor size, organ of origin, definition and quality of tumor margins, tumor heterogeneity, calcification, enhancement, ascites, biliary and/or pancreatic ductal dilatation, local invasion, adenopathy, vascular invasion, vascular encasement, metastases, and signal intensity on MR images. Results from 10 CT, seven US, and three MR imaging examinations were reviewed. RESULTS: Five of the 10 tumors were pancreatic; four others appeared to be pancreatic or hepatic. Most had well-defined margins (nine of 10), were heterogeneous (nine of 10), and enhanced (10 of 10). Other findings included calcification (two of 10), biliary and pancreatic ductal dilatation (one of 10), and ascites (three of 10). Hepatic (two patients) and pelvic (two patients) metastases were present. Adenopathy (two patients) and vascular invasion (one patient) were not identified radiologically. Tumors had low to intermediate signal intensity on T1-weighted images and high signal intensity on T2-weighted images. CONCLUSION: Pancreatoblastoma is typically a heterogeneous tumor with well-defined margins that may appear to arise from the pancreas or liver. It may behave aggressively, with localized vascular or bowel invasion or with widespread metastatic disease. Although it is rare, it should be considered in the differential diagnosis of an upper abdominal mass in a child. PMID- 10671597 TI - Detection of small, functional islet cell tumors in the pancreas: selection of MR imaging sequences for optimal sensitivity. AB - PURPOSE: To determine the sensitivity and specificity of magnetic resonance (MR) imaging for depicting pancreatic small, functional islet cell tumors and the minimum number of sequences for expedient diagnosis. MATERIALS AND METHODS: Twenty-eight patients clinically suspected to have functional islet cell tumors underwent T1- and T2-weighted spin-echo (SE) MR imaging with and without fat suppression, T2-weighted fast SE imaging, and spoiled gradient-echo (GRE) imaging before and after injection of gadopentetate dimeglumine. Sensitivity, specificity, and the best and minimum number of sequences for definitive diagnosis were determined. RESULTS: MR images depicted proved islet cell tumors in 17 of 20 patients (sensitivity, 85%). Images were true-negative in eight patients with negative follow-up examination results for more than 1 year. Specificity was 100%; positive predictive value, 100%; and negative predictive value, 73%. Among 20 patients with tumor, T1-weighted SE images with fat suppression and nonenhanced spoiled GRE images each showed lesions in 15 (75%); T2-weighted conventional SE with fat suppression, in 13 (65%); gadolinium enhanced spoiled GRE, in 12 (60%); and T2-weighted fast SE, in seven of 10 patients (70%). CONCLUSION: MR imaging accurately depicts small islet cell tumors. T2-weighted fast SE and spoiled GRE sequences usually suffice. Gadolinium enhanced sequences are needed only if MR imaging results are equivocal or negative. PMID- 10671598 TI - Anatomy of the right anterosuperior area (segment 8) of the liver: evaluation with helical CT during arterial portography. AB - PURPOSE: To evaluate the segmental anatomy of the right anterosuperior area (segment 8) of the liver by using helical computed tomography during arterial portography (CTAP). MATERIALS AND METHODS: Twenty-seven patients without lesions at segment 8 underwent helical CTAP. Three-dimensional portograms were reconstructed to verify the course of the portal veins. The number of subsegmental branches, in addition to the branching point and the distribution in segment 8, was assessed. RESULTS: In 25 (93%) patients, the dorsal branch of segment 8 gave rise to dorsally directed branches posterior to the right hepatic vein. In only four (25%) of 16 patients in whom the medial branch of segment 8 arose near the porta hepatis, the long paracaval portal branch of the caudate lobe extended upward above the interval between the middle and right hepatic veins. CONCLUSION: In most of the patients, the dorsal branches of segment 8 supplied the dorsocranial area of the right lobe posterior to the right hepatic vein. The paracaval portion of the caudate lobe was limited to below the interval between the middle and right hepatic veins in the majority of patients who showed medial branches of segment 8 arising near the porta hepatis. Recognition of this vascular anatomy is clinically important for preoperative evaluation of hepatic tumors in segment 8 because it may contribute to a safer surgical approach. PMID- 10671599 TI - Diagnosis of gastric cancers: comparison of conventional radiography and digital radiography with a 4 million-pixel charge-coupled device. AB - PURPOSE: To evaluate the differences in accuracy and observer performance at conventional radiography and at digital radiography with a 4 million-pixel charge coupled device (CCD) for the diagnosis of gastric cancers. MATERIALS AND METHODS: A prospective study was performed of 225 patients with suspected gastric cancer who were referred to our hospital from January 1997 through February 1997. One hundred twelve patients were examined at conventional radiography and 113 were examined at digital radiography, and 24 and 27 patients had gastric cancer, respectively. Six radiologists interpreted the images, with attention to tumor findings. They were blinded to the clinical details, and their interpretations were rated against those of three other radiologists who examined the patients and who were aware of the clinical information such as endoscopic features and/or histopathologic findings in biopsy specimens. Receiver operating characteristic (ROC) analysis was used to compare the differences in observer performance for the diagnosis of gastric cancers at conventional radiography and at digital radiography. RESULTS: The overall sensitivity was 64.6% at conventional radiography versus 75.3% at digital radiography (P =. 287); specificities were 84.5% and 90.5%, respectively (P =.011); and the positive predictive values were 53.1% and 71.3%, respectively (P =.036). ROC analysis clearly showed higher diagnostic performance at digital radiography than at conventional radiography. CONCLUSION: The data demonstrate the high diagnostic value of digital radiography with a 4 million-pixel CCD for gastric cancers. The technique has considerable potential as an alternative to conventional gastrointestinal radiography. PMID- 10671600 TI - MR imaging for the preoperative planning of sphincter-saving surgery for tumors of the lower third of the rectum: use of intravenous and endorectal contrast materials. AB - PURPOSE: To evaluate the value of magnetic resonance (MR) imaging with a flexible surface coil in predicting the resectability of tumors in the lower rectum and the feasibility of sphincteral salvage. MATERIALS AND METHODS: In a prospective study, 61 patients with histologically proved primary adenocarcinoma of the lower or middle third of the rectum (<12 cm from the pectinate line) were examined at double-contrast-material-enhanced MR imaging with a circular polarized flexible surface coil. RESULTS: Assessment of anal sphincteral infiltration at MR imaging was excellent, with a specificity of 98% and a sensitivity of 100%. In the determination of tumor infiltration into adjacent organs (T4), the specificity was 100%, and the sensitivity was 90%, with surgical and histologic findings as the standards. While MR imaging showed negative nodes in 40 patients (stage N0 at MR imaging), histologic examination showed negative nodes in 27 patients and positive nodes in 34. At MR imaging, sensitivity was 68%, and specificity was 24%. CONCLUSION: While preoperative staging at MR imaging according to the TNM system still has limited value and accuracy, MR imaging provides the surgeon with valuable information regarding the presence of sphincteral invasion and the surrounding structures in patients with cancers in the lower third of the rectum. PMID- 10671601 TI - Ileal endometriosis: radiographic findings in five cases. AB - PURPOSE: To determine the radiographic findings in five patients with ileal endometriosis. MATERIALS AND METHODS: A search of radiology files revealed five patients with surgically proved endometriotic implants in the ileum at enteroclysis (three patients), at small-bowel follow-through (one patient), and at double-contrast barium enema study (one patient). The radiographic findings were reviewed retrospectively. Clinical, surgical, and histopathologic findings were also reviewed. RESULTS: All five patients were nulliparous women (mean age, 34.4 years; age range, 28-41 years). Four patients presented with abdominal and/or pelvic pain, but only one of these four had cyclic pain that coincided with menstruation. Barium studies revealed endometriotic implants in the terminal ileum within 10 cm of the ileocecal valve in four patients and in the mid-ileum in one. The radiographic findings consisted of extrinsic mass effect with variable spiculation and tethering of folds in two patients, annular lesions with spiculated folds and abrupt or tapered borders in two, and a plaque-like lesion in one. In four patients who underwent double-contrast barium enema studies, associated endometriotic implants were found in the rectosigmoid colon. CONCLUSION: Ileal endometriosis usually involves the terminal ileum within 10 cm of the ileocecal valve and manifests as a spectrum of findings on barium studies. Ileal endometriosis should therefore be considered when these findings are present in young, nulliparous women with abdominal or pelvic pain. PMID- 10671602 TI - Isolated infarction of the cecum: CT findings in two patients. AB - Colonic ischemia isolated to the cecum is a rare entity. The authors evaluated two patients who underwent computed tomography (CT) because appendicitis was suspected at clinical examination. CT findings were suggestive of isolated cecal ischemia or infarction. Surgical-histopathologic findings helped confirm the presumptive CT diagnoses. Isolated cecal infarction should be included in the differential diagnosis of acute right lower quadrant pain. PMID- 10671603 TI - Mucosal detail at CT virtual reality: surface versus volume rendering. AB - PURPOSE: To evaluate computed tomographic virtual reality with volumetric versus surface rendering. MATERIALS AND METHODS: Virtual reality images were reconstructed for 27 normal or pathologic colonic, gastric, or bronchial structures in four ways: the transition zone (a) reconstructed separately from the wall by using volume rendering; (b) with attenuation equal to air; (c) with attenuation equal to wall (soft tissue); (d) with attenuation halfway between air and wall. The four reconstructed images were randomized. Four experienced imagers blinded to the reconstruction graded them from best to worst with predetermined criteria. RESULTS: All readers rated images with the transition zone as a separate structure as overwhelmingly superior (P <.001): Nineteen cases had complete concurrence among all readers. The best of the surface-rendering reconstructions had the transition zone attenuation equal to the wall attenuation (P <.001). The third best reconstruction had the transition zone attenuation equal to the air attenuation, and the worst had the transition zone attenuation halfway between the air and wall attenuation. CONCLUSION: Virtual reality is best with volume rendering, with the transition zone (mucosa) between the wall and air reconstructed as a separate structure. PMID- 10671604 TI - Case 23(1) PMID- 10671605 TI - Diagnosis please. Case 19: enteroliths in a Meckel diverticulum. PMID- 10671606 TI - Primary hyperaldosteronism (Conn syndrome): MR imaging findings. AB - PURPOSE: To describe the magnetic resonance (MR) imaging features of the adrenal glands in primary hyperaldosteronism and assess MR imaging in the detection and characterization of aldosterone-producing adenoma (APA). MATERIALS AND METHODS: The authors retrospectively reviewed the cases of 20 patients (13 female and seven male patients; age range, 14-67 years; median age, 46 years) with primary hyperaldosteronism who underwent 1.5-T MR imaging between 1995 and 1998. All patients underwent transverse T1- and T2-weighted imaging, and chemical shift imaging was performed in 17 patients. Imaging results were correlated with findings at biochemical testing, venous sampling, or surgery. RESULTS: Among the 20 patients, 10 (50%) had APA and 10 (50%) bilateral adrenal hyperplasia (BAH). In the detection of APA, MR imaging had a sensitivity of 70%, specificity of 100%, and accuracy of 85%. APAs (mean size, 20 x 16 mm) were iso- or hypointense relative to the liver on T1-weighted images and slightly hyperintense on T2 weighted images. With chemical shift imaging, the signal intensity decreased on the out-of-phase images in six of seven (86%) patients with APA and in eight of nine (89%) patients with BAH. CONCLUSION: MR imaging has a high specificity in the detection of APA. As with nonhyperfunctioning adenoma, APA and BAH show evidence of intracellular lipid at chemical shift imaging. PMID- 10671607 TI - Cerebellar and frontal lobe hypoplasia in fetuses with trisomy 21: usefulness as combined US markers. AB - PURPOSE: To confirm that cerebellar hypoplasia is ultrasonographically recognizable in second-trimester fetuses with Down syndrome and determine whether the combination of frontal lobe shortening and cerebellar hypoplasia is superior to either measurement alone as a marker of this abnormality. MATERIALS AND METHODS: The frontothalamic distance (FTD) and transcerebellar diameter (TCD) were measured in 52 middle-trimester fetuses with euploid karyotypes and in 52 fetuses with Down syndrome. Receiver operating characteristic (ROC) curves were constructed with various thresholds for observed-to-expected ratios (O/Es) of the FTD, TCD, and average of these two parameters. RESULTS: The area under the average ROC curve, 0.80, was greater than that for either the FTD alone (0.75) or the TCD alone (0.76). At a 6% false-positive rate, the sensitivity for the detection of Down syndrome obtained with the average parameter was 34% better than that obtained with only the FTD and 12% better than that obtained with only the TCD. With an O/E threshold of 0.92 for the average parameter, an odds ratio of 16.3 and positive predictive value of 12.7% in the high-risk population were achieved. CONCLUSION: Although both measurements are individually statistically significant, the combination of TCD and FTD measurements may be superior to the use of either parameter alone as a marker of trisomy 21. PMID- 10671608 TI - Cartilaginous tumors: fast contrast-enhanced MR imaging. AB - PURPOSE: To differentiate between benign and malignant cartilaginous tumors with fast contrast material-enhanced magnetic resonance (MR) imaging. MATERIALS AND METHODS: In 37 patients, fast contrast-enhanced MR images were obtained in eight enchondromas, 11 osteochondromas, and 18 chondrosarcomas. Start of enhancement early, within 10 seconds after arterial enhancement; delayed, between 10 seconds and 2 minutes; late, after 5 minutes on spin-echo images-and progression of enhancement were represented with three types of time-signal intensity curves. Findings were correlated with the surgical specimen in 27 cases, curettage material in three cases, and biopsy combined with long-term follow-up findings in seven cases. RESULTS: Start of enhancement and the combination of start and progression of enhancement correlated significantly (P <.001) with benign and malignant tumors. Early enhancement was seen in chondrosarcoma, not seen in enchondroma, and seen in osteochondroma only when growth plates were unfused. The sensitivity was 89%, specificity 84%, positive predictive value 84%, and negative predictive value 89%. Differentiation of malignancy from benignity on the basis of early and exponential enhancement was possible with a sensitivity of 61%, specificity 95%, positive predictive value 92%, and negative predictive value 72%. CONCLUSION: Preliminary results show that in the adult population fast contrast-enhanced MR imaging may assist in differentiation between benign and malignant cartilaginous tumors. PMID- 10671609 TI - Rates and correlates of discomfort associated with mammography. AB - PURPOSE: To explore the rates and correlates of discomfort at mammography in asymptomatic women aged 50-74 years from six San Diego, Calif, mammography facilities. MATERIALS AND METHODS: Subjects (N = 1,800) completed a 43-item telephone interview approximately 3 weeks after screening mammography. Bivariate associations between variables were analyzed with chi(2) analysis. Logistic regression was used to assess the independent predictors of discomfort at mammography while controlling for all other factors. RESULTS: Nine hundred thirty three (52%) women reported moderate to extreme discomfort at mammography. Discomfort was not related to the intention to undergo future mammography (P =.95). Factors that were significantly associated with discomfort in multivariate analyses were facility (P <.001), satisfaction with care (P <.04), and perception of the technologist's "roughness" (P <.001). CONCLUSION: Discomfort, although not related to the intention to undergo future mammography, had a relatively high incidence. PMID- 10671610 TI - Gynecomastoid hyperplasia: imaging findings in six patients. AB - This case series describes the radiologic appearances of gynecomastoid hyperplasia of the breast in our experience. The clinical histories, breast images, and histopathologic findings in six women were reviewed. At mammography, there was no abnormality in two women, an enlarging asymmetric density in three women, and a nodule in one woman. Breast ultrasonography showed a hypoechoic nodule in one woman. Gynecomastoid hyperplasia has a varied radiologic appearance. PMID- 10671611 TI - Bovine type I collagen as an endovascular stent-graft material: biocompatibility study in rabbits. AB - PURPOSE: To study the biocompatibility of a bovine type I collagen preparation as a material for small-vessel stent-grafts in rabbits. MATERIALS AND METHODS: A composite nitinol-collagen endovascular stent-graft with a 4-mm inner diameter was deployed in the abdominal aorta in nine rabbits. Angiography was performed, and the rabbits were sacrificed at 1, 2, and 7 days and at 1 and 3 months. The portion of the aorta containing the stent-graft was excised and was histologically evaluated. RESULTS: All stent-grafts were patent at all time points. On days 1, 2, and 7 after implantation, scattered red and white blood cells adhered to the stent-graft. At 1 month, the stent-graft was endothelialized and was infiltrated with fibroblasts that deposited collagen within the interstices of the implanted collagen material. At 3 months, there was additional collagen deposition within the interstices of the stent-graft that did not narrow the lumen of the stent-grafts. CONCLUSION: Type I collagen as a intravascular stent-graft material is biocompatible for at least 3 months in rabbits. It is rapidly endothelialized and does not cause reactive stenosis. As a versatile and biocompatible polymer, collagen is potentially useful in the construction of endovascular stent-grafts for use in human arteries. PMID- 10671612 TI - Effect of barium sulfate on wound healing in the gastrointestinal tract of the rat. AB - PURPOSE: To study the effect of barium sulfate on wound healing in the gastrointestinal tract of the rat. MATERIALS AND METHODS: Sixty rats weighing approximately 320 g were divided into four groups: Fifteen control rats had gastric, small-bowel, and colonic incisions; 15 rats had gastric incision; 15 rats had small-bowel incision; and 15 rats had colonic incision. Barium sulfate was placed into the incision before closure in all rats except those in the control group, and the effects were documented clinically and histopathologically for 3 months. Autopsy was performed in five rats from each group at 1, 4, and 12 weeks. The incisions in the rats receiving barium sulfate were compared with those in the control rats. RESULTS: There was no difference in the clinical course (weight gain, activity, and viability) between the control and experimental groups. Early and late autopsy findings and histopathologic grading of healing and inflammatory response were similar for both the control and experimental groups. CONCLUSION: Under the conditions of this study, the effect of barium sulfate on visceral transmural wound healing in the gastrointestinal tract of the rat was minimal. PMID- 10671613 TI - Tumoral distribution of long-circulating dextran-coated iron oxide nanoparticles in a rodent model. AB - PURPOSE: To investigate the accumulation and cellular uptake of long-circulating dextran-coated iron oxide (LCDIO) particles in malignant neoplasms in vivo. MATERIALS AND METHODS: A gliosarcoma rodent model was established to determine the distribution of a model LCDIO preparation in tumors. LCDIO accumulation in tissue sections was evaluated with multichannel fluorescence microscopy with rhodaminated LCDIO, green fluorescent protein as a tumor marker, and Hoechst 33258 dye as an intravital endothelial stain. Uptake into tumor cells was corroborated with results of immunohistochemical and cell culture uptake experiments. The effect of intratumoral LCDIO uptake on magnetic resonance (MR) imaging signal intensity was evaluated with a 1.5-T superconducting magnet. RESULTS: Tumoral accumulation of LCDIO was 0.11% +/- 0.06 of the injected dose per gram of tissue in brain tumors and was sufficient for detection at MR imaging. In tumor sections, LCDIO was preferentially localized in tumor cells (49.0% +/- 4.6) but was also taken up by macrophages in tumors (21.0% +/- 3.1) and by endothelial cells in the areas of active angiogenesis (6.5% +/- 1.4). In cell culture, LCDIO uptake was strongly correlated with growth rate of tumor cell lines. CONCLUSION: Tumoral LCDIO accumulation was not negligible and helped explain MR imaging signal intensity changes observed in clinical trials. Microscopically, LCDIO accumulated predominantly in tumor cells and tumor associated macrophages. Uptake into tumor cells appeared to be directly proportional to cellular proliferation rates. PMID- 10671614 TI - Optimal section spacing in single-detector helical CT. AB - To define the section spacing that maximizes longitudinal resolution without needless section overlap, the optimal percentage of overlap was computed theoretically and expressed as a constant relative to the effective section thickness. For imaging applications that require maximal longitudinal resolution, single-detector helical computed tomographic images should be reconstructed with at least 60% overlap relative to the effective section thickness. PMID- 10671615 TI - US with extended field of view: phantom-tested accuracy of distance measurements. AB - To evaluate the accuracy of distance measurements obtained with the extended field-of-view (FOV) software of a commercially available ultrasonographic scanner, two custom-designed phantoms that allowed scanning of flat and curved surfaces were used. Five hundred forty measurements of various known distances in the phantoms were made by three examiners using various transducers. Although minor differences were observed between operators and transducers, 99.4% (537 of 540) of the distance measurements were accurate within plus or minus 4%. This extended-FOV technology provides accurate measurements of large objects in vitro. PMID- 10671616 TI - Lesions entirely removed during stereotactic biopsy: preoperative localization on the basis of mammographic landmarks and feasibility of freehand technique- initial experience. AB - Seven patients with mammographic lesions entirely removed at percutaneous core needle biopsy that required wider excision underwent freehand localization of the site of the prior lesion with orthogonal and reproducible mammographic landmarks to guide needle placement. Successful excision was accomplished in all cases, as evidenced by similar histopathologic findings, fibrin bands or collagen, and core needle biopsy tract at microscopy. PMID- 10671617 TI - Head and neck tumors: fractionated frameless stereotactic interstitial brachytherapy-initial experience. AB - The authors used a frameless stereotactic navigation system, the Vogele-Bale Hohner head holder, and a targeting device to reproducibly position brachytherapy needles for fractionated interstitial brachytherapy in 12 patients with inoperable cancers of the head and neck. In all cases, deviations of the needle relative to the planned position were within 1-15 mm depending on the location of the tumor. PMID- 10671618 TI - Helical CT cholangiography with oral cholecystographic contrast material. AB - Twenty asymptomatic volunteers underwent helical computed tomographic (CT) cholangiography 10-12 hours after ingesting iopanoic acid. Three observers assessed the images for the extent of bile duct visualization and image quality. The common bile duct and common hepatic duct were adequately visualized in 19 (95%) subjects. Helical CT cholangiography with oral cholecystographic contrast material is feasible and deserves further clinical studies. PMID- 10671619 TI - Potential hazards in the use of tungsten mechanical detachable coils. PMID- 10671620 TI - Proper CT viewing. PMID- 10671621 TI - Compression of the celiac trunk by the median arcuate ligament. PMID- 10671622 TI - Crescent sign origin and the thrombus-to-lumen ratio in abdominal aortic aneurysm. PMID- 10671623 TI - Guide wire-assisted placement of non-end-hole nasoenteric feeding tubes. PMID- 10671624 TI - Abstracts of current literature PMID- 10671625 TI - Renal nutrition in the new millennium. PMID- 10671626 TI - Further analysis in renal nutrition. PMID- 10671627 TI - The best way to manage hypertension after renal transplantation. AB - Hypertension in renal allograft recipients is a common problem arising from multiple factors, including peripheral vascular damage caused by pretransplant hypertension, side effects of immunosuppressive medications, allograft dysfunction, renal artery stenosis, recurrent glomerulonephritis, synthesis of vasoconstrictive hormones by the native kidneys, and excessive dietary salt intake. Identification of modifiable factors causing hypertension and concurrent medical conditions, and measurement of glomerular filtration rate, cyclosporine/tacrolimus blood levels, and magnitude of proteinuria are essential to tailor treatment for an individual patient. Lifestyles that exacerbate hypertension should be modified. For pharmacological therapy, diuretics and calcium channel blockers are first-line agents in patients on cyclosporine shortly after transplant. Angiotensin-converting enzyme inhibitors are good choices for patients with significant proteinuria. Reduction of immunosuppression will improve hypertension in some patients, but entails a potential risk of rejection or graft loss. Angioplasty is necessary in patients with a functionally significant stenosis of the allograft renal artery. Other patients on maximal medical therapy may benefit from native nephrectomy. PMID- 10671628 TI - Protein-energy malnutrition as a risk factor for increased morbidity in long-term hemodialysis patients. AB - This prospective nonintervention single-center study was undertaken to investigate the role of protein-energy malnutrition (PEM) as a risk factor for morbidity in patients on long-term hemodialysis. Thirty-seven patients from the renal unit of Tygerberg Hospital, Tygerberg, South Africa, were studied for a mean period of 26 months. Morbidity was the main outcome and was defined as the number of hospitalizations and days of hospitalization per patient per year. Investigations included 4-monthly determinations of interdialytic protein catabolic rate (PCR), dietary intake of protein and energy, blood levels of albumin and urea, lymphocyte count, adequacy of dialysis (Kt/V), body weight, intradialytic weight loss, fat mass (FM), fat-free mass (FFM), body mass index (BMI), and bone-free arm muscle area (BF-AMA). A PEM composite score was derived from postdialysis serum albumin, BF-AMA, FM, FFM, and BMI. All-cause morbidity as defined by number of hospitalizations (see text for other definitions of morbidity) showed a significant correlation with the mean and baseline PEM score (P <.01), and a negative correlation with predialysis and postdialysis serum albumin (P <.05) and age (P <.05). There was no significant relationship with PCR, percentage intradialytic weight loss, Kt/V, reuse of dialyzer, period on maintenance hemodialysis, sex, race, and type of dialyzer membrane. When "only infection-related" morbidity was considered, the factors that showed a significant correlation were the mean (P <. 001) and baseline PEM score (P <.01), and percentage intradialytic weight loss (P <.01). There was no significant deterioration in the nutritional status of patients followed up for at least 24 months. It is concluded that infection-related morbidity was associated most strongly with the PEM score and the percentage intradialytic weight loss. The results suggest that PEM is one of the important contributing factors to morbidity, possibly via an effect on the immune system and infection. PMID- 10671629 TI - Selenate-supplemented nutritional formula increases plasma selenium in hemodialysis patients. AB - OBJECTIVE: The purpose of this study was to determine the short-term effect of feeding selenium-supplemented formulas on the selenium status of end-stage renal disease patients on hemodialysis. DESIGN AND SETTING: The prospective, randomized, single-blind study of parallel design was conducted at three hemodialysis clinics. PATIENTS: A total of 79 hemodialysis patients were randomly assigned into one of three groups. INTERVENTION: Liquid nutritional formula supplemented with either selenite (28 microg Se/8 oz, n = 26), selenate (28 microg Se/8 oz, n = 26), or nonfortified (7 microg Se/8 oz, n = 27) was fed to hemodialysis patients as their sole source of nutrition for 14 days. MAIN OUTCOME MEASURE: Plasma and red blood cell (RBC) selenium and glutathione peroxidase (GPX) activities were measured in predialysis blood both before (day 1) and after (day 8) a 7-day baseline period, and after subjects received the formula as the sole source of nutrition (approximately 35 kcal/kg/d) for 14 days (day 22). RESULTS: Selenium intake (Mean +/- SEM, microg/d) was 134 +/- 9, 140 +/- 9, and 35 +/- 2 for patients receiving selenite-, selenate-, or non-supplemented formula, respectively. On day 22, plasma selenium (micromol/L) was greater (P <.032) in the selenate-supplemented group (1.5 +/- 0.1) compared with the nonsupplemented group (1.2 +/- 0.1), but not compared with the selenite supplemented group (1.4 +/- 0.1). Plasma GPX activity was 44% to 60% that of healthy controls and not different among groups. RBC selenium and GPX activities were within the normal range and were not different among groups. CONCLUSION: The results of this study indicate that a liquid formula supplemented with selenium as selenate is successful at maintaining selenium concentrations within normal range, as well as significantly increasing plasma selenium levels compared with nonsupplementation. PMID- 10671630 TI - Oral vitamin intake in children receiving long-term dialysis. AB - OBJECTIVE: To evaluate dietary and oral supplement vitamin intake in children submitted to dialysis (peritoneal dialysis and hemodialysis). DESIGN: Prospective clinical trial in a 12-month follow-up period. SETTING: Children with end-stage renal disease (ESRD) who attended the pediatric nephrology clinic of Universidade Federal de Sao Paulo-Escola Paulista de Medicina (UNIFESP-EPM), Sao Paulo, Brazil. PATIENTS: Thirty children (18 girls, 23 in peritoneal dialysis, 7 in hemodialysis) with age 9.3 +/- 7.4 years. INTERVENTION METHODOLOGY: Six successive assessments of both anthropometric indexes and 3-day dietary diaries in children receiving a daily dose of oral water-soluble vitamin supplement. MAIN OUTCOME MEASURES: Anthropometric indexes (weight/age [W/A], height/age [H/A], midarm muscle area/age [MAMA/A], and fat area/age [FA/A]) and dietary adequacy-% recommended dietary allowance (RDA) (computerized nutritional analysis from 3-day dietary intake diary). RESULTS: Anthropometric indexes analysis showed that 53% of children were <-2.0 standard deviation score (SDS) of W/A, 63% were <-2.0 SDS of H/A, and 43.3% were <-1.65 SDS of MAMA/A, suggesting growth deficit and low muscle wasted. Total caloric intake was lower than 100% of RDA in 90% of children. Dietary intake of water-soluble vitamins was <100% of RDA in the majority of children, as follows: vitamin C (24/30), B1 (28/30), B2 (22/30), B3 (27/30), B6 (26/30), B12 (1/30), pantothenic acid (24/30), and folic acid (9/30). The combined dietary and vitamin supplement intake resulted in excessive oral intake for almost all the vitamins. CONCLUSION: Dietary intake of water-soluble vitamins is lower than the RDA in the majority of children with ESRD; supplementation is necessary to reach the RDA. The use of the available vitamin supplement resulted in vitamin intakes that exceeded the RDA for almost all of the vitamins. However, we do not know if these intakes exceeded the children's requirements, nor whether they had any clinically significant harmful effects. PMID- 10671631 TI - Can low-fat/cholesterol nutrition counseling improve food intake habits and hyperlipidemia of renal transplant patients? AB - OBJECTIVE: To assess the impact of low-fat/cholesterol nutrition counseling on food intake habits and blood lipid levels of renal transplant patients. DESIGN: Prospective practice-based outcome study. SETTING: Acute care hospital post-renal transplant outpatient clinic. PATIENTS: Forty-three renal transplant patients not on lipid-lowering medications referred to the renal dietitian for low fat/cholesterol nutrition counseling between September 1994 and September 1997. INTERVENTION: Individual assessment and counseling using the Healthy Heart Nutrition Guidelines Step 1 diet (<30% of total calories from fat, <300 mg cholesterol, and <10% of total calories from saturated fatty acids). MAIN OUTCOME MEASURES: Three-day food records precounseling and 3-day food records (n = 13) or descriptive intake changes (n = 30) postcounseling (time interval: 2 to 8 months). Fasting/random serum total cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein-cholesterol (LDL-C), as available. RESULTS: Total cholesterol (n = 43) and LDL-C (n = 22) decreased significantly (0. 54 mmol/L P <.000 and 0.53 mmol/L P <.000, respectively). There were no significant changes in HDL-C and triglycerides. Twenty percent of patients (n = 43) reached target levels of total cholesterol <5.2 mmol/L, and 35% of patients (n = 22) reached target levels of LDL-C <3.4 mmol/L. Percentage of total calories from fat decreased significantly (7.58% P <.03). Descriptive intake changes of lower fat choices were reported in the no post-food records group. CONCLUSION: Patients can make changes in food intake habits after nutrition counseling. Serum lipid levels can improve after nutrition counseling, but many patients may still require lipid lowering medications to reach target levels. Nutrition counseling should be considered for the initial treatment of hyperlipidemia in renal transplant patients. PMID- 10671632 TI - The effects of megestrol acetate on nutritional parameters in a dialysis population. AB - OBJECTIVE: To examine the effect of megestrol acetate on nutritional parameters in a hemodialysis population. DESIGN: Prospective case studies of hemodialysis patients. SETTING: A freestanding, nonprofit, hemodialysis unit. SUBJECTS: Seventeen patients were studied. They were included regardless of gender, age, or cause of renal disease. They had to be on dialysis for at least 2 months, had a serum albumin <3.5 g/dL for these 2 months, and had to be at high nutritional risk. There were 8 women and 9 men. Ages were 44 to 87 years. Eight were diabetics, and 9 were nondiabetics. INTERVENTIONS: Megestrol acetate 400 mg orally twice daily was prescribed, and patients were studied for 6 months. OUTCOME MEASURES: Pre-evaluation and postevaluation were performed by patient questionnaire, Subjective Global Assessment (SGA), dry weight, and anthropometric measurements. Monthly laboratory monitoring included albumin, prealbumin, blood urea nitrogen (BUN), cholesterol, triglycerides, carbon dioxide, platelets, hematocrit, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gammaglutamyl transpeptidase (GGT), lactate dehydrogenase (LDH), alkaline phosphatase, and glucose. Glycohemoglobin and hemoglobin A1c were monitored in diabetic patients. RESULTS: Three patients were able to take megestrol acetate for 5 to 6 months. They reported improved appetite and showed an increase in dry weight. The annualized mortality rate was about 59%. Side effects included diarrhea, confusion, hyperglycemia, headaches, dizziness, and elevated LDH. CONCLUSION: Megestrol acetate may help stimulate appetite in the hemodialysis patient, but it is risky and must be monitored closely. Eight hundred milligrams per day is probably too large a dose for the end-stage renal disease (ESRD) patient. PMID- 10671633 TI - Computers and the Internet: tools for lifelong learning. AB - Mastery of computer technology and the resources it brings to renal dietitians will be essential for future practice. The Internet, probably the most valuable resource computers bring to clinical practice, is a potential tool to fulfill goals of lifelong learning. A basic introduction to the Internet and a review of search engines and successful search strategies are provided. Major flaws of the Internet include uncertain quality and accuracy of many materials it provides. Guidelines to evaluate World Wide Web (WWW) materials are similar to guidelines dietitians use to assess written materials. The identification of certain anchor WWW sites with reliable information is a key to successful information retrieval on the Internet. PMID- 10671635 TI - Message from the chairperson PMID- 10671634 TI - Grocery list for people on hemodialysis! (Food your patients CAN eat). PMID- 10671636 TI - Duplicated regions of AF-4 intron 4 at t(4;11) translocation breakpoints. AB - BACKGROUND: AF-4 is a common partner gene of MLL. AF-4 breakpoints occur in introns, but most AF-4 introns are uncharacterized. METHODS AND RESULTS: We cloned AF-4 intron 4 and examined the frequency of breakpoints in this intron. The 5.8-kb intron is rich in repeat sequences and was the site of translocation in 3 of 17 leukemias with t(4;11). We cloned the der (11) and der (4) breakpoints and isolated the fusion transcripts in the cell line MV4-11 and in a de novo acute lymphoblastic leukemia (ALL). Both translocations joined MLL intron 6 and AF-4 intron 4. In MV4-11, 249 bases from AF-4 were present in both derivative chromosomes, indicating duplication. In the de novo ALL, duplication of 446 bases from MLL and AF-4 occurred. Reciprocal fusion transcripts were expressed. CONCLUSIONS: Intronic sequence of AF-4 is useful for molecular diagnosis of t(4;11). Duplicated intronic regions suggest staggered chromosomal breakage. PMID- 10671638 TI - Pharmacogenetics becomes pharmacogenomics: wake up and get ready. PMID- 10671637 TI - Improved detection of human immunodeficiency virus type 1 variants by analysis of replicate amplification reactions: relevance to studies of human immunodeficiency virus type 1 vertical transmission. AB - BACKGROUND: Human immunodeficiency virus type 1 (HIV-1)-infected individuals typically harbor mixtures of HIV-1 variants. For HIV-1 transmission studies, methods used for genotypic analysis should reliably detect variant mixtures. Such studies typically analyze complementary DNAs (cDNAs) from a single polymerase chain reaction (PCR) amplification. This approach may fail to detect variant mixtures in some samples because of analytic bias. METHODS: To evaluate the impact of analytic bias on the detection of HIV-1 variants, we analyzed samples from a mother and infant known to contain both subtypes A and D HIV-1. The env third variable region of HIV-1 gp120 (V3 region) was amplified and cloned in five replicate experiments using a single plasma sample from each individual. Ten cDNAs from each experiment were analyzed. RESULTS: The subtype mixture was detected in only four of 10 amplification experiments (three of five for the mother and one of five for the infant). Sequencing of uncloned PCR products showed that a single subtype, either A or D, was preferentially amplified in each experiment. However, the subtype mixture was detected for each sample by analyzing the five replicate experiments as a group. CONCLUSIONS: This shows that mixtures of HIV-1 variants may be more readily detected when replicate amplification reactions are analyzed. This approach may be useful for characterizing HIV-1 variants for studies of HIV-1 transmission. PMID- 10671639 TI - Impact of pharmacogenomics on the clinical laboratory. AB - Clinical pharmacogenomics promises to increase the safety and efficacy of drug prescription, decrease the incidence of adverse drug reactions, help improve public health, and presage in an era of personalized, predictive, and prophylactic medicine. Clinical pharmacogenomics stands to be broad based and include the following laboratory components: new and expanded pharmacogenetic tests, disease profiles, chemopredictive testing, and risk profiling. There is a growing body of evidence that variable drug responsiveness is caused by polymorphisms within multiple genes, protein products of which are involved in critical metabolic and/or physiologic pathways relevant for drug action. Different pharmacogenomic approaches will be used to discover a new generation of unique and highly predictive pharmacogenetic tests that the clinical laboratory will employ to help identify patient responder populations. Disease profiling and chemopredictive testing will routinely be applied to accurately screen for disease and help guide therapeutic course of action. A growing number of risk profiling tests will assist in predicting a patient's predisposition to disease. Clinical pharmacogenomics stands to become the basis for the new millennium's practice of medicine and have a profound impact on the clinical laboratory. PMID- 10671640 TI - Populations and genetic polymorphisms. AB - BACKGROUND: Population frequencies of many polymorphic genes of pharmacogenetic interest depend on race or ethnic specificity. Association of these genes with person-to-person differences in drug effectiveness (hypersensitivity or resistance) and drug toxicity may also depend on the racial or ethnic characteristics of a population. Information about ethnic specificity is an integral part of pharmacogenetics because it can suggest a starting point for further study of these traits, tailoring drug therapy to the individual patient, and rational development and clinical trials of new drugs. Ethnic specificities of several medically important metabolic traits serve to illustrate these ideas. Among the traits considered is primaquine sensitivity, a sex-linked trait attributed to glucose-6-phosphate dehydrogenase deficiency that mainly affects males among African, Mediterranean, and Oriental people. Additional examples include the remarkable sensitivity of the Japanese to alcohol (ethanol) compared with whites; the ethnic specificity of the cytochrome P-450 enzyme CYP2D6* (debrisoquine/sparteine) polymorphism that results in poor, extensive, and ultrarapid metabolizers of at least 30 drugs; the CYP2C19* (mephenytoin) polymorphism that accounts for variable metabolism of proguanil, omeprazole, and certain barbiturates; and the polymorphic (NAT2*) acetylation of hydrazine and aromatic amine drugs, such as isoniazid, hydralazine, and sulfasalazine. PMID- 10671641 TI - Single-nucleotide polymorphisms, haplotypes, and their relevance to pharmacogenetics. AB - Recognition that there is a vast quantity of human genetic variation has had a pervasive impact on modern medicine, facilitating the identification of scores of genes that underlie monogenic clinical disorders, as well as genes involved in complex disease processes. The next logical step for human genetics is the exploration and elucidation of genes involved in differential pharmacological response: responders, nonresponders, and those with adverse side effects. An understanding of the role that genes have in pharmacological response is the cornerstone of personalized medicine. Pharmacogenetic activities have swiftly embraced these tenets, leading to a proliferation of resources and approaches meant to enable and expedite targeted drug discovery and development. To realize the potential of these efforts, it will be necessary to incorporate a better understanding of the population genetic and evolutionary processes that have shaped genetic variation in modern humans. This article introduces these concepts to provide context and guidelines for the use of this variation (primarily single nucleotide polymorphisms and haplotypes) in pharmacogenetics. PMID- 10671642 TI - Mining the Swedish clinical archives to develop pharmacogenomic tests. AB - Eurona Medical is a Swedish company that develops diagnostic tests to predict response to drugs or treatment. Sweden offers unparalleled retrospective clinical data resources, with epidemiological registers and collections of tissue samples built up over decades. Efficient pharmacogenomic research can be performed using these registers and sample collections in collaboration with experienced medical researchers. Eurona's tests are based on Genetic Signatures, groups of polymorphic, polygenic genomic positions linked to and therefore predictive of drug response. These are elucidated from complex data sets using unique applications of multivariate and combinatorial statistics and a multigenic approach. The company develops tests applicable to current medical practice and is preparing to launch its first within the hypertension field. Quality control (including ISO9001 certification) and clinical regulatory compliance are applied throughout all programs to produce data that can be directly translated into clinical tests. PMID- 10671643 TI - Inherited variations in drug-metabolizing enzymes: significance in clinical oncology. AB - Pharmacogenetics has emerged as a novel and challenging area of interest in oncology. Cancer chemotherapy is characterized by major intersubject variability in tumor responses and host toxicity. This variation may be caused by genetic differences in the enzymes involved in the metabolism of anticancer agents. Anticancer agents, such as 6-mercaptopurine, 5-fluorouracil, and irinotecan, have a narrow therapeutic index that can sometimes result in severe life-threatening toxicities. The impact of polymorphisms in metabolizing enzymes (thiopurine S methyltransferase, dihydropyrimidine dehydrogenase, and uridine diphosphate glucuronosyltransferase) that participate significantly in the disposition of these anticancer agents is discussed. PMID- 10671644 TI - Apolipoprotein E: a pharmacogenetic target for the treatment of Alzheimer's disease. AB - BACKGROUND: The discovery that the apolipoprotein E4 (apoE4) allele is strongly linked to both sporadic and familial late-onset Alzheimer's disease (AD) raises the possibility that a dysfunction of the lipid transport system could seriously affect lipid homeostasis in the brain. We recently proposed that the abnormally low concentrations of apoE observed in the brains of apoE4 AD subjects could compromise cholesterol, fatty acid, and phospholipid transport in the central nervous system. This, in turn, would indirectly impair the cholinergic system, which, in contrast to other neurotransmitters in the central nervous system, relies heavily on lipids to synthesize acetylcholine. Several independent investigators have now confirmed the original observation of an inverse relationship between apoE4 allele copy number and residual brain choline acetyltransferase activity and nicotinic-receptor binding sites in the brains of subjects with AD. More importantly, it has been shown that the presence of the apoE4 allele differentially affects the quality and size of drug responsiveness in subjects with AD treated with cholinomimetic and noncholinomimetic agents. We also examine the role of apoE as a potent therapeutic target for AD. PMID- 10671645 TI - Technologies for detecting genetic polymorphisms in pharmacogenomics. AB - BACKGROUND: Pharmacogenomics is an emerging scientific discipline examining the genetic basis for individual variations in response to therapeutics. METHODS AND RESULTS: Genetic polymorphisms are a major cause of individual differences in drug response. Metabolic phenotyping can be accomplished by administering a probe drug or substrate and measuring the metabolites and clinical outcomes. However, this approach tends to be labor intensive and requires repeated sample collection from the individual being tested. Alternatively, genotyping allows determination of individual DNA sequence differences for a particular trait. Commonly used genotyping methods include gel electrophoresis-based techniques, such as polymerase chain reaction (PCR) coupled with restriction fragment length polymorphism analysis, multiplex PCR, and allele-specific amplification. Fluorescent dye-based high-throughput genotyping procedures are increasing in popularity, including oligonucleotide ligation assay, direct heterozygote sequencing, and TaqMan (Perkin Elmer, Foster City, CA) allelic discrimination. High-density chip array and mass spectrometry technologies are the newest advances in the genotyping field, but their wide application is yet to be developed. Novel mutations/polymorphisms also can be identified by conformation based mutation screening and direct high-throughput heterozygote sequencing. CONCLUSIONS: Rapid and accurate detection of genetic polymorphisms has great potential for application to drug development, animal toxicity studies, improvement of human clinical trials, and postmarket monitoring surveillance for drug efficacy and toxicity. PMID- 10671646 TI - Clinical, genetic, and pharmacogenetic applications of the Invader assay. AB - The Invader technology has been developed for the detection of nucleic acids. It is a signal amplification system able to accurately quantify DNA and RNA targets with high sensitivity. Exquisite specificity is achieved by combining hybridization with enzyme recognition, which provides the ability to discriminate mutant from wild-type at ratios greater than 1/1000 (mutant/wt). The technology is isothermal and flexible and incorporates a homogeneous fluorescence readout. It is therefore readily adaptable for use in clinical reference laboratories, as well as high-throughput applications using 96-, 384-, and 1,536-well microtiter plate formats. The molecular mechanism of the system and specific applications for use in clinical and research laboratories are described. These include direct analysis of unamplified human genomic DNA to detect mutations and single nucleotide polymorphisms associated with factor V Leiden, factor II, cystic fibrosis, and apolipoprotein E, and gene expression assays that quantify messenger RNA levels in cells using direct lysates. PMID- 10671647 TI - Pharmacogenetics in the practice of laboratory medicine. AB - BACKGROUND: The clinical laboratory forms an essential bridge between fundamental discoveries in biological sciences and their transition into effective medical practice. The genetic basis for individuality in drug metabolism and response is the result of a finite number of inherited sequence variants (alleles) of genes encoding drug-metabolizing enzymes and drug receptors. Pharmacogenetics (PG) links differences in gene structure with pharmacological differences in pharmacokinetics and pharmacodynamics. The next step in the process of applying PG (or pharmacogenomic) information to individualized therapeutic management is dissemination of this information to practicing physicians by clinical laboratorians. Transitioning PG analysis into clinical practice will require professionals in laboratory medicine to identify relevant polymorphisms, develop sensitive and specific testing strategies, and, in conjunction with physicians and pharmacologists, communicate interpretive guidelines regarding appropriate indications for testing and rational dose adjustment. We review these concepts and provide examples of how PG can be applied to support therapeutic decision making. PMID- 10671648 TI - The use of aptamers in large arrays for molecular diagnostics. AB - BACKGROUND: Aptamers are single-stranded oligonucleotides derived from an in vitro evolution protocol called systematic evolution of ligands by exponential enrichment (SELEX). They bind tightly and specifically to target molecules; most aptamers to proteins bind with Kds (equilibrium dissociation constant) in the range of 1 pM to 1 nM. METHODS AND RESULTS: The SELEX protocol has been automated; therefore, hundreds to thousands of aptamers can be made in an economically feasible fashion. Blood and urine can be analyzed on chips that capture and quantitate proteins. SELEX has been adapted to the use of 5-bromo (5 Br) and 5-iodo (5-I) deoxyuridine residues. These halogenated bases can be specifically cross-linked to proteins. Selection pressure during in vitro evolution can be applied for both binding specificity and specific photo-cross linkability. These are sufficiently independent parameters to allow one reagent, a photo-cross-linkable aptamer, to substitute for two reagents, the capture antibody and the detection antibody, in a typical sandwich array. After a cycle of binding, washing, cross-linking, and detergent washing, proteins will be specifically and covalently linked to their cognate aptamers. CONCLUSIONS: Because no other proteins are present on the chips, protein-specific stain will now show a meaningful array of pixels on the chip. Learning algorithms and retrospective studies should lead to a robust, simple, diagnostic chip. PMID- 10671649 TI - Whither the chips may fall? PMID- 10671650 TI - Overestimation of HFE C282Y homozygous hemochromatosis prevalence as the result of a common primer-binding site polymorphism. PMID- 10671651 TI - Gynecologic cancer: evolving management issues. Introduction. PMID- 10671652 TI - Complete surgical staging of early endometrial adenocarcinoma: optimizing patient outcomes. AB - Endometrial adenocarcinoma is the most common gynecologic malignancy. Strategies for treatment of this disease should not only emphasize quality of care resulting in cure of disease, but also use health care resources in the most efficient manner possible. Based on available data, we recommend that all patients with the diagnosis of endometrial carcinoma undergo complete surgical staging with lymph node dissection. Radiation therapy is reserved only for patients with evidence of extrauterine disease. This approach maximizes the amount of information available for treatment planning and offers the potential therapeutic advantage of lymph node dissection. Additionally, in a cost analysis, this approach appears to be the most cost-effective. PMID- 10671653 TI - Complete surgical staging in endometrial cancer provides prognostic information only. AB - Endometrial cancer is the most common gynecologic malignancy in the United States. Surgical resection provides the best chance for cure and guides the potential need for adjuvant therapy. Controversy exists regarding the extent of surgical staging, in particular, the necessity and rationale for removing or sampling the draining lymphatics. An emerging practice in the United States is to perform more aggressive lymph node dissections. The data indicate that surgical staging provides prognostic information only and may indirectly influence survival only through guiding adjuvant therapy. PMID- 10671654 TI - The role of radiotherapy for high-risk endometrial cancer. AB - High-risk endometrial cancer comprises an uncommon group of tumors, which includes pathological stage III adenocarcinoma and all stages of papillary serous carcinoma. Optimal management of this class of malignant female genital neoplasms is surgical resection, including debulking of any gross abdominopelvic disease. This article analyzes the literature concerning the use of adjunctive radiotherapy. The data presented suggest that postoperative whole abdominal radiotherapy may improve outcome in selected subsets of patients within this high risk group. Future clinical investigations will greatly benefit from the anticipated published results of two completed prospective cooperative group clinical trials that involve whole abdominal irradiation. PMID- 10671655 TI - Adjuvant chemotherapy for high-risk endometrial cancer. AB - Identification of histopathologic factors that predict the risk of tumor recurrence allows for selection of women with endometrial cancer who might benefit from adjuvant therapy. Most studies of adjuvant treatment have focused on external-beam irradiation or oral progestational agents and have failed to document a survival advantage for treated patients. Although recurrent or metastatic endometrial tumors often respond to salvage treatment with cytotoxic agents, there is relatively little experience with postoperative systemic chemotherapy used in an adjuvant setting. A few nonrandomized trials-using doxorubicin/platinum-based regimens-have suggested that adjuvant chemotherapy may be beneficial in some patient subsets. Data from larger-scale, randomized trials do not exist. Additional clinical experience is needed before a definite role for adjuvant chemotherapy can be established. PMID- 10671656 TI - Tailoring radiation to the extent of disease for uterine-confined endometrial cancer. AB - More than 50 years ago, endometrial cancer was found to be sensitive to radiation, and adjuvant radiation was observed to decrease the incidence of pelvic recurrences. Over the last 2 decades, substantial progress has been made in the understanding of prognostic factors for survival and patterns of disease recurrence for patients with endometrial cancer. Few randomized trials have been done because of the relatively few patients who are at risk of recurrence and the strong bias of many oncologists toward the use of adjuvant radiation. Principles guiding treatment recommendations are based predominately on retrospective publications containing variance in pathological evaluation, surgical evaluation, and patient selection. Preliminary analysis of a randomized Gynecologic Oncology Group trial is reviewed. Optimal therapy for many patients remains to be better defined. PMID- 10671657 TI - Optimizing radiation parameters for cervical cancer. AB - The treatment of cervical cancer has become increasingly sophisticated with evidence-based guidelines generated from randomized trials directing combined modality programs. The radiotherapeutic guidelines have been derived largely from single institutional experiences coupled with data from the Patterns of Care Studies. The design of external-beam fields has improved because of a better understanding of the anatomy of the cervix, uterus, parametrium, and draining lymph nodes from surgical, lymphangiogram, computed tomography scan, and magnetic resonance imaging series. An improvement in survival and local control with dose intensity (a reduction in overall treatment time and increase in overall dose) has been shown for cervical cancer, especially for locally advanced disease, and these series are highlighted. The use of intracavitary radiation, which is technically accurate, dramatically improves outcome and allows for dose intensity. Alternative brachytherapy techniques, such as high-dose-rate, and interstitial, are discussed. Ways to improve the therapeutic ratio for radiation, including biological factors, are reviewed. More research is necessary to understand the complex dose distribution of external-beam and intracavitary brachytherapy and its relationship to tumor control. PMID- 10671658 TI - Concurrent chemotherapy and radiation for locally advanced cervical cancer: the new standard of care. AB - Radical pelvic irradiation has constituted the definitive therapy for patients with large cervical cancers. No substantial improvements have been made in treatment outcomes. In the past year, however, a series of large, well-conducted randomized trials has evaluated the role of concurrent chemotherapy with pelvic irradiation in cervical cancer. These trials include definitive treatment of patients with stage IB2 to IVA disease and adjuvant treatment after radical surgery in stage IB2-IIA disease. Five trials have shown a consistent 30% to 50% reduction in the risk of death from disease when concurrent chemotherapy is used. Questions still remain as to what constitutes the best chemotherapy dose and schedule. In all of the positive trials, cisplatin was used, but three also used 5-fluorouracil. The level of survival improvement that occurs when chemotherapy is added to optimal irradiation and whether patients with stage IIIB and IVA benefit are also unclear. Improvements in survival rates for patients with solid tumors occur slowly. Based on the evidence, it is likely that concurrent chemotherapy with radiation will become the new standard of care for bulky and advanced cervix cancer. PMID- 10671659 TI - Adjuvant therapy for high-risk, early stage cervical cancer. AB - The identification of various pathologic risk factors after primary surgical management of early stage cervical cancer portends a higher rate of relapse and decreased survival. Historical attempts to improve outcome focused mainly on the use of adjuvant pelvic radiation, with limited success overall. Analysis of patterns of failure after radical hysterectomy led to better stratification of patients into risk groups and incorporated testing of systemic agents in those considered at high risk of distant failure. Two recently reported randomized, clinical trials have greatly advanced our understanding of the role of postoperative therapy in cervix cancer. In patients with positive nodes, the use of combined adjuvant chemotherapy and radiation significantly improves relapse free survival and overall survival, compared with radiation alone. For node negative patients with other primary tumor risk features, pelvic radiation significantly improves relapse-free survival, compared with no further therapy. An observed improvement in survival for irradiated patients awaits statistical confirmation after maturation of the data. Further improvements in adjuvant therapy for high risk, early stage cervical cancer will come from enhanced definition of prognostic variables, better patient selection, and refinements in both local and systemic therapies. PMID- 10671660 TI - Integrating radiation therapy in the curative management of ovarian cancer: current issues and future directions. AB - Although important advances in surgery, chemotherapy (CT), and radiation therapy (RT) have been made, overall survival for patients with ovarian cancer (OC) has not changed significantly. Despite its long history in the treatment of OC and its proven curative role in patients with microscopic or minimal residual disease, the proper role of RT in the management of OC is not clearly established. Although the use of primary adjuvant RT (whole abdominal irradiation) has declined in the last 15 years, there has been a resurgence of interest in RT as part of a combined modality approach and as salvage therapy for patients with small-volume persistent disease after primary cytoreductive surgery and platinum-based CT. This article reviews the evidence supporting the use of RT alone or combined with chemotherapy as primary adjuvant therapy or in the salvage setting. Current issues in the radiotherapeutic management are discussed along with ideas for future clinical research directions. PMID- 10671661 TI - Differential effects of retinoids on nitric oxide production by promonocytic U937 cells and ZR-75-1 human breast cancer cells. AB - We demonstrated the presence of inducible and endothelial nitric oxide synthases in histiocytic lymphoma U937 cells by staining with anti-iNOS and anti-eNOS antibodies. We also investigated the effects of retinol and retinoic acid on nitric oxide production by both U937 cells and ZR-75-1 human breast cancer cells. U937 cells which had been treated with either retinol or retinoic acid (10-10-10 6 M) exhibited no significant difference in nitric oxide secretion into conditioned medium. Conversely, for ZR-75-1 cells, both retinol and retinoic acid (10-10-10-6 M) caused a significant (p<0.001) increase in the amount of nitrite secreted. Our results indicate that retinoid induced growth inhibition of breast cancer cells is associated with an increase in NO production, however, an increase in NO synthesis does not mediate retinoid induced differentiation of U937 cells. PMID- 10671662 TI - A novel monoclonal antibody, H9, directed against the core protein of MUC1 mucin. AB - MUC1 mucin is a target protein for many monoclonal antibodies. Human MUC1 detected by a murine anti-KL-6 monoclonal antibody that recognizes a sialylated carbohydrate chain has been designated KL-6/MUC1. Given the heterogeneous antigenicity of KL-6/MUC1, we established a new murine monoclonal antibody, H9, that reacts with epitope DTRP (Asp-Thr-Arg-Pro) peptides within the immunodominant region of the tandem repeat of MUC1 mucin. The reactivity of the H9 antibody differs from that of other previously reported antibodies that recognize the tandem repeat region of MUC1. Immunohistochemical experiments indicate that the reactivity of the H9 antibody is similar to that of other antibodies directed against MUC1 core proteins. A new cancer-associated protein detected by a sandwich assay using the H9 antibody as a catcher and the KL-6 antibody as a tracer is designated HK9. Serum HK9 levels showed a high expression level in lung cancer: 51% (19/37 cases) for adenocarcinoma, 39% (11/28 cases) for squamous cell carcinoma, and 67% (10/15 cases) for small cell carcinoma. The HK9 expression in lung cancer increased with cancer progression. These findings suggest monoclonal antibody H9 to be a novel antibody that reacts with an epitope within the tandem repeat region of MUC1, and that the cancer-associated antigen HK9 may have useful tumor-associated properties. PMID- 10671663 TI - pS2 expression as a possible diagnostic marker of colorectal carcinoma in ulcerative colitis. AB - This study was performed to evaluate the significance of pS2 and MUC1 expressions in ulcerative colitis (UC)-associated colorectal neoplasias. Tissues were collected from 6 patients with UC-associated colorectal neoplasias treated surgically. Specimens were 13 adenocarcinomas, 40 dysplasias (20 high-grade dysplasias, 20 low-grade dysplasias), and 60 normal mucosae. Tissues were also collected from patients without UC treated surgically or endoscopically. pS2, p53, and MUC1 expressions were examined immunohistochemically and compared. The K ras codon 12 mutation was investigated by single-strand conformation polymorphism analysis. In patients with UC, the incidence of pS2 expression was significantly higher (p<0.01) in adenocarcinomas than it was in dysplasias, and no pS2 expression was seen in normal mucosae. p53 overexpression was detected in 50% (10/20) even in low-grade dysplasias. MUC1 expression was seen only in invasive carcinomas, but it was seen in 100% of cases (3/3). K-ras gene mutations were detected in 2 (20%) of 10 carcinomas. In low and high-grade dysplasias, the incidences of pS2 expression were significantly (p<0.01) lower than the incidences of p53 overexpression, however, in UC-associated carcinomas there was no significant difference; pS2 expression and p53 overexpression were detected in 13 of 13 (100%) cases and in 12 of 13 (92%) cases, respectively. These results suggest that p53 overexpression may be a diagnostic marker of neoplasia, and that pS2 expression may be a diagnostic marker of colorectal carcinoma in case of UC. PMID- 10671664 TI - A favorable impact of preoperative FPLC chemotherapy on patients with gastric cardia cancer. AB - The aim of this study is to evaluate the effects of preoperative chemotherapy with fluorouracili polyphase liposome composita pro orale (FPLC) on the tumour cells and the survival rate of the patients with gastric cardia cancer. Sixty patients with gastric cardia cancer were randomly divided into two groups. Thirty patients were treated with FPLC prior to surgical resection, the other 30, as controls, did not receive the preoperative chemotherapy. Pathological responses of the tumours to the FPLC chemotherapy were determined by gross and microscopic assessments of tumour size, tumour emboli, cell degeneration and necrosis. Expressions of nm23 and CD44 were detected by flow cytometry. All patients were followed up to 5 years. In the FPLC-treated patients, the tumour size (p<0. 01), the number of tumour emboli (p=0.04) and the intensity of CD44 expression (p<0.001), were significantly reduced, while cell degeneration (p<0.001), necrosis (p<0.01) and the expression of nm23 (p<0.001) were increased, when compared with those observations seen in the controls. The postoperative 5-year survival rate was 40% in the FPLC-treated group and 23% in the controls (p=0.17). Preoperative FPLC chemotherapy might improve the survival rate of patients with gastric cardia cancer by inhibiting tumour proliferative, invasive and metastatic activities, and stimulating the patient's immune system. PMID- 10671665 TI - Is a woman's date of birth related to her risk of developing breast cancer? AB - Some previous reports have shown seasonality in the date of birth in patients with breast cancer, others have not. In this study of 1,110 women with unilateral breast carcinoma significantly more cases had been born in the first than the second half of the year. This finding could, however, not be linked to the later development of breast cancer as their case-matched controls showed a similar trend. These results underline the importance of using relevant control material in such studies. PMID- 10671666 TI - A simultaneous monitoring of Lens culinaris agglutinin A-reactive alpha fetoprotein and des-gamma-carboxy prothrombin as an early diagnosis of hepatocellular carcinoma in the follow-up of cirrhotic patients. AB - To elucidate the risk factors for developing hepatocellular carcinoma (HCC) during the follow-up of patients with liver cirrhosis (LC), outpatients with LC were examined periodically by means of serum biochemical assessments, ultrasonography, and computed tomography. Risk factors for HCC were statistically analyzed. We also examined an efficacy of Lens culinaris agglutinin A-reactive profiles of alpha-fetoprotein (AFP-L3%) and des-gamma-carboxy prothrombin (DCP) value using a highly sensitive DCP determination kit (ED036) for the early recognition of HCC. The AFP-L3% and the ED036 value were retrospectively determined with stored serum samples. HCC was diagnosed in 21 of the 78 patients with LC during the follow-up period (mean follow-up period: 42 months). The estimated cumulative incidence of HCC was 25% with 3 years and 48% with 5 years. The most significant risk factor for the development of HCC in LC patients was found to be the mean serum AFP concentration from the year before the HCC detection (p=0.02). At the time of the recognition of HCC, the positive rates of the tumor markers were: serum AFP concentration 14%, serum DCP value 5%, AFP-L3% was 33%, and that of ED036 43%. The positive rate in collaborative use of AFP-L3% and ED036 was 67%. The simultaneous determination of the AFP-L3% and the ED036 value was shown to be effective for the early detection of HCC. PMID- 10671667 TI - In vivo chemosensitivity of human malignant cystosarcoma phyllodes xenografts. AB - Malignant cystosarcoma phyllodes (MCSP) is a rare breast tumor. Chemotherapeutic regimens for treatment of MCSP have not been established. We previously established an MCSP xenograft line MC-3-JCK. In this study, we established a new MCSP xenograft line, MC-10-JCK, by serial transplantation in nude mice. We studied the chemosensitivity of these two MCSP tumor xenografts to anticancer drugs in vivo. We also examined the expression of multidrug resistance-related proteins such as p-glycoprotein (Pgp) and multidrug resistance-associated protein (MRP) by immunohistochemical analysis. These two xenografts were sensitive to doxorubicin, vincristine and cyclophosphamide in vivo. Immunohistochemically, clinical specimens and xenografts were negative for Pgp and MRP expression. These results are consistent with the chemosensitivity of human MCSP to lipophilic anticancer compounds. PMID- 10671668 TI - Induction of p53-dependent apoptosis in vivo by nedaplatin and ionizing radiation. AB - p53 protein expression, apoptosis and growth delay induced by nedaplatin, a novel platinum compound, were investigated in vivo, and compared with those induced by ionizing radiation. A human ependymoblastoma with wild-type p53 was transplanted subcutaneously to the thighs of nude mice. The incidences of p53 protein-positive cells and apoptosis in tumors increased following exposure to ionizing radiation. In tumors treated with nedaplatin, they also increased, but the incidences of p53 protein-positive cells and apoptosis induced by 32 mg/kg nedaplatin, 1/2 LD50, were lower than those induced by 1 Gy irradiation. However, growth-delay assay showed no significant difference between the efficacy of 32 mg/kg nedaplatin and that of 1 Gy irradiation. These results suggest that the main antineoplastic activity caused by nedaplatin may be mediated through different mechanisms than those of the p53-dependent early apoptosis. PMID- 10671669 TI - Comparison of effects of doxorubicin and radiation on p53-dependent apoptosis in vivo. AB - The effects of doxorubicin and radiation on apoptosis, p53 expression, and tumor growth in human tumor xenografts were investigated. Human ependymoblastoma (NNE), primitive neuroectodermal tumor (YKP), glioblastoma (KYG) and small cell lung carcinoma (GLS) that are all transplantable to nude mice were treated with doxorubicin (8 mg/kg) or radiation (1 Gy). The histological study was performed by using TUNEL and p53 staining. Cytotoxic effects of doxorubicin and radiation were compared with no-treatment group by the growth curves and apoptotic index of tumor to each treatment. In NNE with wild-type p53, doxorubicin induced growth delay of tumors (tumor volume doubling time; 13.7+/-3.3 days in control group vs 30.4+/-1.5 days in doxorubicin group), but no growth delay of tumors in KYG and GLS with mutant type p53. While radiation-induced apoptosis appeared most frequently at 6 h after irradiation, doxorubicin-induced apoptosis had a tendency to appear later. Furthermore, although the frequency of doxorubicin-induced apoptosis was lower than that of apoptosis by 1 Gy irradiation, apoptotic cells appeared for many hours after the treatment. Doxorubicin-induced apoptosis may be correlated with p53 phenotype because apoptosis was induced only in tumor with wild-type p53, but it appeared less frequently and later than radiation-induced apoptosis. PMID- 10671670 TI - Perioperative quantitative analysis of cytokeratin 20 mRNA in peripheral venous blood of patients with colorectal adenocarcinoma. AB - Hematogenous dissemination is a significant short-coming of colorectal carcinoma treatment. To screen patients with high risk for such blood-borne metastasis, we previously developed a highly sensitive system for the detection of cytokeratin 20 (CK-20) mRNA in blood. For a more practical application, we improved this system by making it quantitative and capable of analyzing peripheral venous blood for the detection of perioperative changes in CK-20 mRNA. CK-20 mRNA was not always detected in the preoperative blood, even in patients in an advanced stage, but it was identified without fail in intra- and post-operative blood. In addition, more copies of CK-20 mRNA were observed in the intra-operative blood than in pre- and post-operative blood. This study suggests that analysis of perioperative changes may provide important information for the precise evaluation of hematogenous dissemination and of the effect of surgical maneuvers on recurrence. PMID- 10671671 TI - Expression of estrogen and progesterone receptors in carcinomas of the female breast in Tanzania. AB - In Africa breast cancer has been reported to occur frequently in young females and to show an aggressive histological and clinical picture, suggesting that this malignancy might have a different biology from this disease in Western females. To investigate this, the present study assessed by immunohistochemistry the expression of estrogen receptors (ER) and progesterone receptors (PgR) in 60 fresh frozen breast cancer tissues from indigenous Tanzanian patients. This prospective study collected tissues from routine patients treated at the Muhimbili Medical Center, Dar es Salaam, Tanzania. These markers have not been previously investigated in indigenous sub-Saharan females with breast cancer. Patients in this study expressed lower frequencies of ER (33%) and PgR (18%) as compared to literature reports including those about African-Americans. Expression of these markers, however, correlated with the demographic, clinical and histological characteristics in a similar way as observed elsewhere. A compounding effect of younger patients' age, advanced disease or late stage at hospital presentation and race in this geographical region could be responsible for the poor expression of hormonal receptors in the majority of patients as observed in this study. A surprising finding was that the proportion of hormonal receptor positive tumors increased with disease duration. In view of the low frequency of expression of hormonal markers, only 26.7% of the patients would be expected to benefit from hormonal therapy based on their expression of the hormone receptors. There is great need to undertake an inter-African study that would evaluate the hormonal status of more African women with breast cancer in different geographical regions of sub-Saharan Africa and document the true picture of their hormonal status. The outcome of these results could be important for treatment strategies for the second most common cancer among African women. PMID- 10671672 TI - Bcl-2 related proteins are dramatically induced at the early stage of differentiation in human liver cancer cells by a histone deacetylase inhibitor projecting an anti-apoptotic role during this period. AB - Expression of the Bcl-2 family members in a human hepatocellular carcinoma cell line (HCC-T) after sodium butyrate-treatment was investigated. Sodium butyrate, a histone deacetylase inhibitor, induced differentiation of the cell line into its normal counterpart without inducing apoptosis at the concentration of 2 mmol/l. Since sodium butyrate has effects on both differentiation and apoptosis, we investigated the expression profile of bcl-2 related genes in HCC-T. The expression of Bcl-2 and Mcl-1/EAT was up-regulated 4-12 h after the treatment while Bcl-XL was up-regulated 2-3 days after the stimulation. On the other hand, the expression levels of Bax protein remained unchanged during differentiation. The HCC-T cells entered a cell cycle arrest at G1 and showed neither cellular fragmentation nor apoptosis during this period, which was concomitantly associated with up-regulated expression of a cell cycle regulator, p21WAF-1. These results demonstrate that induction of anti-apoptotic bcl-2 related proteins at an early stage of differentiation is important for the maintenance of HCC-T cell differentiation by antagonizing pro-apoptotic molecules such as Bax. PMID- 10671673 TI - Experimental study of the effects on apoptosis of docetaxel alone and in combination with irradiation. AB - The effects of docetaxel alone and in combination with irradiation were experimentally investigated in terms of induction of apoptosis. A human ependymoblastoma was transplanted into nude mice, and they were intravenously injected with docetaxel, followed or preceded by irradiation with a single dose of 2 Gy. Apoptosis was less common in the docetaxel-alone group than in the irradiation-alone group. When administration of docetaxel was followed by irradiation, apoptosis was equal to or less common than in the irradiation-alone group. Apoptosis was most common in the irradiation-followed-by-docetaxel group. The incidence of mitosis was lower in the irradiation-alone group and the irradiation-followed-by-docetaxel group than in the docetaxel-alone group and the docetaxel-followed-by-irradiation group. The combined effect of irradiation and docetaxel appears to be useful in inducing apoptosis, but the sequence of irradiation and docetaxel is important. PMID- 10671674 TI - Immunohistochemical study on primary and recurrent tumors in patients with local recurrence in the conserved breast. AB - One hundred and seventy patients received breast-conserving therapy in the Second Department of Surgery, Gunma University School of Medicine. Six (3.5%) out of the 170 patients showed breast recurrence. We investigated the breast recurrent cases clinicopathologically. The age at the initial operation ranged from 38 to 78 (mean 57) years. One patient was clinical stage I and the others were clinical stage II. Surgical margin at the initial operation was negative in two patients and positive in four. Histological type was invasive ductal cancer in all cases. Three patients had lymph node involvement. The interval from the initial operation to breast recurrence ranged from 19 to 68 months. Five cases were nodular type and one was diffuse type of breast recurrence. Histological type of breast recurrence was the same as the initial one. We performed salvage surgery for all breast recurrent patients, mastectomy for four patients and local resection for two. One patient who showed diffuse type of recurrence could not be controlled with any surgical treatment, and later died of breast cancer. We investigated the expression of estrogen receptor, progesterone receptor, pS2, c erbB-2 and p53 on both initial and recurrent specimens of the six patients. The expression of each protein on the recurrent specimens was the same as the initial one. We conclude that breast recurrence after breast-conserving therapy has its origin in the residue of cancer cells at the initial operation, even if surgical margins are histopathologically negative. PMID- 10671675 TI - DNA double-strand break rejoining in human follicular lymphoma and glioblastoma tumor cells. AB - Follicle center cell lymphoma is among the most radioresponsive of human cancers. To assess whether this radioresponsiveness might be a result of a compromised ability of the tumor cells to accomplish the biologically-effective repair of DNA double-strand breaks (DSBs), we have measured i) the extent of the mechanical rejoining of radiation-induced DSBs in biopsy-derived follicle center cell lymphoma cells and ii) the fidelity with which nuclear protein extracts from these cells rejoin restriction enzyme-induced DSBs. Cell suspensions derived from two lymphoma biopsies, designated FCL1 and FCL2, as well as two established human glioblastoma cell lines, M059J and M059K, were exposed to 30 Gy of gamma-rays and evaluated for their ability to rejoin DSBs using a Southern transfer-pulsed-field gel electrophoresis assay. The fidelity of rejoining of restriction enzyme induced DSBs was assessed using a cell-free plasmid reactivation assay. Both lymphoma suspensions rejoined DSBs relatively slowly and exhibited a similar phenotype to the known DSB-rejoining deficient M059J line. The level of DSB mis rejoining in the cell-free plasmid reactivation assay was also similar in M059J and FCL2 cells and was considerably ( approximately 6-fold) higher than in M059K cells. Because of insufficient numbers of cells, we were unable to perform this assay with the FCL1 lymphoma. These limited data suggest that follicle center cell lymphoma cells may be intrinsically deficient in performing the biologically effective rejoining of DSBs. Such a deficiency might contribute to the radioresponsiveness of this disease and may be exploitable in the development of improved treatment strategies, such as radioimmunotherapy. PMID- 10671676 TI - Dose-intensive weekly alternating chemotherapy for patients with small cell lung cancer: randomized trial, can it improve survival of patients with good prognostic factors? AB - We conducted a randomized trial of dose-intensive weekly alternating chemotherapy (CAV/PE-W) and standard alternating chemotherapy (CAV/PE) in small cell lung cancer (SCLC) patients with good prognostic factors. A total of 76 patients with SCLC was randomized. The CAV/PE-W consisted of 4 alternating cycles of cyclophosphamide: 500 mg/m2, doxorubicin: 30 mg/m2, and vincristine: 1 mg/m2 (day 1) and cisplatin: 50 mg/m2 (day 8) and etoposide: 75 mg/m2 (days 8 and 9). The CAV/PE consisted of 2 alternating cycles of cyclophosphamide: 800 mg/m2, doxorubicin: 50 mg/m2, and vincristine: 1.4 mg/m2 (day 1), cisplatin: 100 mg/m2 (day 22) and etoposide: 100 mg/m2 (days 22, 23 and 24). Eligibility criteria were no prior therapy, no active concomitant malignancy, ECOG PS of 0 or 1, age < or =75, adequate hematologic functions and no brain metastasis. The complete response (CR) rate for CAV/PE-W (14/38, 36.8%) was significantly higher than that for CAV/PE (6/38, 15.8%, chi2; p=0. 032). However, the response rate in patients on CAV/PE-W (36/38, 94. 7%) was not significantly higher than the rate for CAV/PE (31/38, 81. 6%, chi2; p=0.076). Progression-free survival for patients on CAV/PE W was significantly longer than that of patients on CAV/PE (41.4 weeks vs. 21.3 weeks, log-rank; p=0.0007, generalized Wilcoxon; p=0.0034) as was overall median survival (67.0 weeks vs. 51.2 weeks, log-rank; p=0.028). Actual dose-intensity of CAV/PE-W was 1.74 times that of CAV/PE. Hematological toxicities were equally frequent and G-CSF contributes to treatment efficacy by allowing administration of dose-intensive chemotherapy. The CAV/PE-W achieved a higher CR rate and longer survival, than the CAV/PE. PMID- 10671677 TI - Retrospective study of the prognostic factors of remission induction for single agent chemotherapy with cisplatin in advanced epithelial ovarian cancer. AB - A retrospective study to explore the prognostic factor was conducted in 39 patients with advanced epithelial ovarian cancer, FIGO stage III-IV, who underwent single-agent adjuvant chemotherapy with cisplatin following primary debulking surgery. The survival rate following the adjuvant chemotherapy was 50.9% after 3 years and 44. 7% after 5 years, with a median survival time of 40.4 months. The actual dose intensity of the cisplatin ranged from 14.38 to 46.30 mg/m2/week, and the total dose/m2 was 143.79 to 645.83 mg/m2. Under these therapeutic conditions, the actual dose intensity was found to correlate with survival time (p<0.05), but there was no correlation between the total dose/m2 and survival time. PMID- 10671678 TI - Genomic DNA analyses of spontaneous hepatocellular carcinomas in LEC rat liver using a new technique. AB - An inbred rat strain, LEC (long evans cinnamon) has been used as a model of human Wilson's disease. This animal suffers from a severe type of hepatitis, the clinical manifestations of which are similar to human fulminant hepatitis for 4-5 months which is caused by accumulation of copper in the liver. The surviving rats develop chronic hepatitis, followed by the development of spontaneous hepatoma. In contrast to studies with hepatocellular carcinomas (HCCs), the studies have great advantages in that the animals have identical genetic background, can be raised under a fixed condition, and the development of HCC is reproducible. We took two HCC samples and analysed their genomic DNA using RLGS (restriction landmark genomic scanning), which involves two-dimensional electrophoresis of genomic DNA allowing the survey of some 1,000 NotI sites throughout the genome. Using this technique, we discovered landmark spots that were either decreased or increased in intensity in HCC and compared them with the RLGS profile obtained from the DNA of control normal LEC rat liver. Approximately 1,300 spots were compared, and the intensity of two spots was found to be decreased about half and one was increased 1.3-1.7 folds. Although the mechanism of these changes and the properties of the changed DNA are yet to be studied, recurrent genomic changes in the LEC rat HCC could prove to be a good model system for elucidating the essential genetic events in association with hepatocarcinogenesis. PMID- 10671679 TI - Prognostic comparison between peripheral and central types of squamous cell carcinoma of the lung in patients undergoing surgical resection. AB - In order to define whether the location of the tumor [peripheral (P) or central (C)] may have some influence on the prognosis for patients with squamous cell carcinoma of the lung, we analyzed 235 patients under 80 years of age (P-group = 129, C-group = 106) who had undergone surgical resection between January 1985 and December 1997. There was no significant difference in the prognosis between the two groups with stages I(0)-IIIB of the disease. We concluded that as a whole the location of the tumor may not have significant influence on the prognosis in patients with squamous cell carcinoma of the lung undergoing surgical resection. PMID- 10671680 TI - Mutational analysis of the CTNNB1 (beta-catenin) gene in human endometrial cancer: frequent mutations at codon 34 that cause nuclear accumulation. AB - Recently, CTNNB1 (beta-catenin) has been found to function as an oncoprotein that works in the Wnt signaling pathway, and mutation of this gene has been reported in various human cancers. In this study, we analyzed 44 endometrial cancers and found somatic missense mutations in five (11%) tumors. Interestingly, four (80%) of the five tumors with mutations would cause amino acid alterations at residues next to Ser 33, one of the targets for phosphorylation of glycogen synthase kinase (GSK)-3beta. The tumors with mutations showed accumulation of the CTNNB1 protein in cytoplasm and nucleus. This is the first report of frequent somatic mutation of the CTNNB1 gene at codons adjacent to those encoding to Ser/Thr residues in endometrial cancer. PMID- 10671681 TI - Exploratory study of effective chemotherapy to clear cell carcinoma of the ovary. AB - Although clear cell carcinoma of the ovary is considered to be a tumor with poor prognosis, the clinical characteristics has not been defined. The aim of this study was to evaluate the response of clear cell carcinoma of the ovary to first and second-line chemotherapy and explore effective chemotherapy. Fifty-three patients with clear cell carcinoma of the ovary were enrolled between 1988 and 1997 at our department. Since taxol was not available in Japan at that time, cisplatin-based combination chemotherapy has been exclusively used as a standard first-line chemotherapy. Retrospective analyses of clinical characteristics and the response to first or second-line chemotherapy were performed. Median age was 52 years (range 27-71 years). Tumors were 34% (18/53) stage I, 19% (5/53) stage II, 38% (20/53) stage III, and 9% (5/53) stage IV. All patients with I or II stage disease had optimal cytoreduction. Out of 25 patients with III or IV stage disease 20% (5/25) had negative residual tumor, 36% (9/25) had <2 cm residual tumor, and 44% (11/25) had >/=2 cm residual tumor. All patients received postoperative platinum-based chemotherapy. Of 23 patients with measurable residual tumor 8.7% (2/23) completely and 13% (3/23) partially responded to first line chemotherapy consisting of cisplatin, adriamycin and cyclophosphamide (CAP) or cisplatin and cyclophosphamide (CP) by CT scan or second look laparotomy. Presence of endometriosis was 55% (29/53) but was not a prognostic factor. Although overall response rate of ovarian clear cell carcinoma to first-line chemotherapy by CAP or CP was about 22%, EP or EJ consisting of etoposide and cisplatin or carboplatin used as a second-line chemotherapy showed 29% response rate, while CPT-P consisting of CPT-11 and cisplatin showed 40% response rate. Clear cell carcinomas were frequently present at early stage, with association of endometriosis and with poor overall prognosis. Although patients with advanced ovarian clear cell carcinoma seemed to have better response to CPT-P than conventional platinum-based chemotherapy, further studies are required with larger number of patients to draw firm conclusions. PMID- 10671682 TI - Clinical significance of serum vascular endothelial growth factor in colorectal cancer patients: correlation with clinicopathological factors and tumor markers. AB - Vascular endothelial growth factor (VEGF) is known as a potent inducer of angiogenesis in various human cancers. Serum VEGF concentrations of colorectal cancer patients was assessed for their clinical significance as a tumor marker. Serum samples were obtained at admission from 24 healthy volunteers and 111 patients with colorectal cancer. Preoperative serum VEGF concentrations, which are significantly higher than those of healthy controls, reflect clinical stage progression, depth of invasion, liver metastasis, lymph node metastasis and lymphatic invasion. Consequently, detection of VEGF could serve as a clinically useful marker for colorectal cancer progression and metastasis independent of other markers. PMID- 10671683 TI - Neoadjuvant chemotherapy for osteosarcoma of the extremities with synchronous lung metastases: treatment with cisplatin, adriamycin and high dose of methotrexate and ifosfamide. AB - We report on the clinical course and outcome of 28 patients, treated at The Istituti Ortopedici Rizzoli between 1995 and 1997 for osteosarcoma of the extremities metastatic to the lung at presentation. The treatment for these patients was the following: primary chemotherapy with cisplatin, adriamycin and high dose of methotrexate and ifosfamide followed by simultaneous resection of primary and metastatic lesions (when feasible), and further chemotherapy. After primary chemotherapy, lung metastases disappeared in 6 patients, whereas metastases in 3 remained surgically unresectable. These 9 patients received surgical treatment of the primary tumor only. In the remaining 19 patients, after chemotherapy, a simultaneous resection of the primary and metastatic tumor was performed. The resection of metastatic lesions was complete in 18 cases and incomplete in one. Three of the 4 patients who did not achieve a tumor-free status died in a few months and one is still alive with uncontrolled disease. With a median follow-up of 32 months (19-43) of the 24 patients who achieved remission, 12 (55%) remained continuously free of disease, 11 relapsed with new metastases and 1 died of chemotherapy-related toxicity. The 2-year DFS and OS were 36% and 53% respectively. These results are much worse than those achieved in 114 contemporary patients with localised disease (2-year DFS: 81%) treated in the same period and they are superimposible to the results achieved in 23 patients previously treated with the same protocol, but with standard dose of ifosfamide (2-year DFS: 32%). However, it must be underlined that, as regards prognosis, patients with metastatic disease at presentation are a hetero-geneous group. The DFS was significantly higher for patients with only one or two metastatic lesions than for patients with 3 or more lesions (2 year DFS: 78% vs. 28%). In 12 of the 19 patients who had a complete simultaneous resection of the primary and metastatic tumor, a strong correlation between the degree of necrosis of the primary and metastatic lesions was found. We conclude that in patients with osteosarcoma of the extremity with lung metastases at presentation: a) the combination of aggressive chemotherapy with simultaneous resection of primary and metastatic tumors works very well only for those patients who present with one or two metastatic nodules whereas for patients with 3 or more pulmonary metastases the prognosis is very poor; b) within the 4-drug regimen used in this study, the increment of ifosfamide dose from 10 g/m2 to 15 g/m2 for cycle does not improve prognosis; c) the strong correlation found between the histologic response of the primary tumor and metastases supports the strategy, largely used nowadays in the neoadjuvant treatment of osteosarcoma, of tailoring postoperative chemo-therapy on the basis of the primary tumor histologic response to preoperative chemotherapy. PMID- 10671684 TI - Antiproliferative action of melatonin on human prostate cancer LNCaP cells. AB - Recent experimental evidence suggests that melatonin, the major pineal hormone, might possess oncostatic properties. The present experiments were performed to verify whether melatonin might modulate the growth of androgen-dependent prostate cancer cells (LNCaP) and to obtain information on its possible mechanism of action. We have shown that melatonin, when given in the nanomolar range, significantly inhibits the proliferation of LNCaP cells; moreover, the pineal gland hormone affects cell cycle distribution by inducing an accumulation of the cells in G0/G1 and a decrease in S phase. To investigate the mechanism of action of melatonin, by RT-PCR analysis we were able to demonstrate the expression, in prostate cancer cells, of a mRNA coding for the membrane Mel1a melatonin receptor. However, by radioreceptor assay, no detectable binding of 2 [125I]iodomelatonin could be observed in membrane preparations from these cells, suggesting that the levels of translation of the mRNA for Mel1a are possibly too low to mediate the antiproliferative action of the hormone. This hypothesis is further supported by the following observations: i) melatonin analogs, specifically acting through membrane receptors (i.e., 2-bromomelatonin), were completely ineffective in modulating prostate cancer cell proliferation; ii) melatonin failed to prevent forskolin-induced cAMP accumulation. These results indicate that melatonin, at nanomolar concentrations, exerts a direct antiproliferative action on androgen-dependent prostate cancer cells, significantly affecting their distribution throughout the cell cycle. Membrane receptors do not seem to be involved in the oncostatic action of the pineal gland hormone. PMID- 10671685 TI - Correlation of immunohistochemical staining and mutations of p53 in human hepatocellular carcinoma. AB - Mutations of the p53 tumor suppressor gene are common in hepatocellular carcinomas (HCCs). Detection of mutations by sequencing provides more information than immunohistochemical staining, but the equipment needed and the time required make it less practical for use in large-scale studies or in studies in developing countries. The degree of correlation between results obtained with these two methods has been studied in various tumors but has not been well-established in human HCCs. Paraffin sections of HCCs of 28 patients from Qidong, China were immunohistochemically stained using monoclonal antibody to p53. In addition, exons 5-8 of the p53 gene were sequenced in these HCCs. Of the 28 HCCs, nine had 0-9% of nuclei stained for p53, and 19 had 50-95% stained. Mutations in p53 exons 5-8 were found in 17/28 (61%) HCCs, including 15 at codon 249 (exon 7), one at codon 198 (exon 6), and one at codon 175 (exon 5). Among these 17 cases with p53 mutations, 16 cases (94%) had 50-95% of nuclei stained. Among 11 HCCs with no mutations by sequencing, 8 were also negative by immunohistochemistry (0-9% of nuclei stained) (73%) (the five HCCs with no staining whatsoever all had wild type p53). Immunohistochemical staining to detect p53 mutations in human HCCs detected most mutations that were detected by sequencing (94% sensitivity, 73% specificity), and this method is therefore suitable when sequencing cannot be performed. PMID- 10671686 TI - Serum interleukin-10 is an independent prognostic factor in advanced solid tumors. AB - Interleukin (IL)-10 is a Th2 type pleiotropic cytokine that has been found to be produced at the tumor site and to be increased in sera of patients suffering from different types of cancer. IL-10 has been shown to hinder a number of immune functions, i.e., T lymphocyte proliferation, Th1 type cytokine production, antigen presentation, and lymphokine-activated killer cell cytotoxicity. To assess its prognostic value, we measured serum levels of IL-10 in 118 patients with advanced solid tumors before treatment, after completion of therapy, and during follow-up. Other prognostic variables, to which IL-10 results were compared, were analyzed as well. IL-10 serum levels were found significantly elevated in cancer patients with respect to healthy controls. Of interest, a significant decrease in IL-10 serum levels was observed in the responder group, whereas a significant increase was recorded in the non-responder group. Using univariate and multivariate analyses, a significant relationship was shown between IL-10 serum levels and both overall survival (OS) and time to treatment failure (TTF). Stepwise regression analysis selected IL-10 serum level, performance status (PS), and stage as the best association of variables with significant impact on OS and TTF. In conclusion, this study shows that IL-10 has an independent prognostic significance in patients with advanced solid tumors and may be useful for monitoring disease progression. PMID- 10671687 TI - Bovine seminal ribonuclease inhibits in vivo growth of human neuroblastoma cells. AB - Bovine seminal ribonuclease (BS-RNase) is a homologue of RNase A with specific antitumor activities. It is selectively toxic for neuroblastoma (NB) cells in vitro with no significant effects on the viability of normal human cells. We evaluated the antitumoral effects of BS-RNase on human NB xenografts from UKF-NB 3 cells in athymic (nude) mice. The efficacy of direct intraneoplastic, subcutaneous and systemic delivery of BS-RNase was explored. Systemic administration of BS-RNase (12.5 mg/kg/day intraperitoneally, for 20 days in the course of four weeks) suppressed tumor growth but was not able to induce any cures. Subcutaneous injections (12.5 mg/kg/day for 20 days in the course of four weeks) and intratumoral BS-RNase treatment using the same schedule resulted in complete tumor regression. During 30 days following cessation of treatment no tumor regrowth was observed and animals were free of tumors. Toxic effects of BS RNase (e.g., on bone marrow and inner organs) were not apparent. This data indicates that BS-RNase fulfills important criteria for a candidate antitumor agent specific for NB. PMID- 10671688 TI - Frequent genotype changes at -308 of the human tumor necrosis factor-alpha promoter region in human uterine endometrial cancer. AB - This study was designed to test the hypothesis that genotypic changes of tumor necrosis factor alpha (TNF-alpha) promoter region are associated with endometrial cancer. TNF-alpha gene encodes a cytokine involved in angiogenesis and oncogenesis of several cancers. Some uncommon alleles are reported to be associated with increased production of TNF-alpha and the onset of various cancers. Studies of such alleles are lacking in endometrial cancer. In this study, we examined the genetic changes at the polymorphic 3 loci of TNF-alpha promoter regions (-238, -308 and 488) in 41 Japanese patients with uterine endometrial carcinomas (UEC). Nine of 41 UEC samples (22%) had genotype changes from GA to A at -308 promoter region of the TNF-alpha gene. There were no genotype changes at -238 and 488 regions of TNF-alpha in UEC samples. The results of our study demonstrate for the first time that genotype changes from GA to A in UEC patients. We conclude that the genotype changes at -308 promoter region of TNF-alpha may play an essential role in the malignant transformation of endometrial cells. PMID- 10671689 TI - The effect of external beam irradiation after endoscopic palliation of esophageal carcinoma. AB - The aim of this study was to verify the value of additional external beam irradiation (EBR) after endoscopic palliation, regarding quality of life and survival rate. From January 1988 to December 1995, 99 patients with esophageal carcinoma (squamous cell carcinoma 61; adenocarcinoma 38) were reviewed, there were 84 males (mean age: 67 years) and 16 females (mean age: 65 years). Seventeen patients were in stage IIb, 45 stage III and 37 patients in stage IV. HDR brachyradiotherapy (mean: 14.7 Gy) was carried out in all patients. Additional EBR (mean: 47.8 Gy) after endoluminal palliation was done in 51 cases. At 6 months follow-up swallowing of a semi-solid diet at least was possible in all patients and dysphagia was found with significant difference in favour to EBR only in stage IV (p=0.011). The Karnovsky performance status showed a difference in favour of EBR for stage III and IV (p=0.040 and p=0. 049, respectively). The median overall survival for EBR compared to no EBR was 10 and 7 months, with a 12 months survival rate of 60% and 16% (p=0.0012). However, considering different stages and EBR versus no EBR a significant difference in survival could only be found for stage IIb (p=0.031), a trend in favour of EBR could be found for stage III (p=0.0985) and stage IV (p=0.0543). Tumor regrowth 6-12 months after treatment occurred in 31 cases and was successfully treated with Nd-YAG laser in 25 and stenting in 6 cases. Postirradiation fibrotic stenosis occurred in 12 cases. Improved survival rates after additional EBR can only be expected in stage IIb. However, in case of advanced esophageal carcinoma and fair performance status, EBR after endoluminal palliation help to maintain quality of life. PMID- 10671690 TI - p53 inactivating mutations in chinese breast carcinomas. AB - While previous reports on breast cancers indicate that Caucasian women have a low frequency of p53 mutations, higher frequencies of mutations are reported in some Japanese populations. There are few reports regarding p53 mutations in Chinese breast carcinomas. Using a previously established sensitive p53 yeast functional assay, we screened 23 Chinese breast carcinomas for mutations. The p53 was mutated in 5/23 (21.7%) specimens. Two of these mutations were detected in exon 5 and one was detected in each of exons 6, 7 and 8. All of these mutations have previously been shown to be mutated in Caucasian and Japanese breast cancers, but two have not previously been observed in Chinese breast cancers and one has also not been observed in Japanese. PMID- 10671692 TI - High fat diet elevates the activity of inducible nitric oxide synthase and 1,2 dimethylhydrazine-induced aberrant crypt foci in colon of rats. AB - Rats were examined for the effect of dietary level of fat (beef tallow or safflower oil) on colonic activity of inducible nitric oxide synthase (iNOS) which has been suggested to be associated with colon carcinogenesis. In experiment 1, feeding high fat diets (20%) for 3 weeks caused higher activity of the iNOS compared to feeding low fat diets (5%). In experiment 2, rats were fed 20% or 5% beef tallow diet for 11 or 32 days, and given an injection of a carcinogen, 1,2-dimethylhydrazine on day 4. The activity of colonic iNOS and the development of colonic aberrant crypt foci were enhanced by high fat diet. PMID- 10671691 TI - Susceptibility to the induction of glutathione S-transferase positive hepatic foci in offspring rats after gamma-ray exposure during gestation. AB - Glutathion S-transferase positive (GST-P+) hepatic foci development was used as a means of determining whether the offspring of gestating maternal rats, which were subjected to genetically-damaging levels of gamma-ray radiation, were more susceptible to the development of cancer after treatment with diethylnitrosamine (DEN), a known carcinogen. A single dose of 10, 30, 60, and 90 rads involving whole body exposure to gamma-rays was given to pregnant rats at day 14, and during postnatal week 4. DEN was intraperitoneally injected to their offspring twice in one week. Thirteen weeks after birth, the rats were sacrificed. Irradiation of maternal rats with 30 rad gamma-rays before mating significantly increased both the incidence and the size of GST-P+ foci in the livers of both male and female pups, when combined with DEN treatment, whereas other dose levels had no such effect. Using a rat-liver model, the results of this study indicate that a low dose of radiation during the embryonic stage increases the susceptibility to carcinogens. In addition, under certain circumstances low doses of radiation, an externally applied cancer-inducing stimulus, may increase the likelihood of cancer, whereas higher doses may not. PMID- 10671693 TI - Efficacy and safety profile of amifostine in the preoperative combined therapy of esophageal cancer patients. AB - The aim of this study was to assess the protective effect and the safety profile of amifostine in 16 esophageal cancer patients undergoing neoadjuvant chemo radiation therapy (group A) compared to 21 matched patients (group B), treated with the same schedule without receiving amifostine, and considered as controls. Haematological and extra-haematological toxicity were evaluated according to WHO criteria and considered as result of amifostine activity. The bone marrow toxicity was globally lower in group A than in group B. We recorded 4 cases of mucosities in group B compared to 1 case in group A. amifostine-related side effects were few (2 cases of hypotension and 1 of vomiting), mild, and well controlled. In conclusion, amifostine seems to be effective and safe when used as protective agent also in esophageal cancer. PMID- 10671694 TI - Cytogenetic and molecular abnormalities in astrocytic gliomas (Review). AB - In recent years, there have been great advances in our understanding of the genetic events and the molecular biology of human brain gliomas. Cytogenetic information has suggested that a pattern of non-random abnormalities involving numerical deviations such as the gain, partial deletion, or total loss of chromosomes as well as translocations and structural rearrangements of certain chromosome lesions are characteristic features for some tumors. In addition, the somatic activation of cellular oncogenes and inactivation of tumor suppressor genes represent important genetic alterations leading to progressive disorder of normal cellular growth control mechanisms. This review describes the abnormal chromosomal and molecular abnormalities that occur during formation of brain tumors of astrocytic origin, particularly fibrillary astrocytic neoplasms. The most frequent genetic alterations include inactivation of the p53, p16, Rb and PTEN genes, and overexpression of the CDK4, EGFR and VEGF genes. Other less well defined abnormalities include aberrations in chromosomes 1, 9, 10, 11, 19 and 22. PMID- 10671695 TI - Giant cell tumor of tendon sheath, tenosynovial giant cell tumor, and pigmented villonodular synovitis: defining the presentation, surgical therapy and recurrence. AB - Giant cell tumor of the tendon sheath (GCTTS), tenosynovial giant cell tumor (TGCT), and pigmented villo-nodular synovitis (PVNS) are the common names for a group of rare proliferative disorders that involve synovial joints and tendon sheaths. Considerable confusion exists about the surgical treatment and diagnosis of these disorders. This review evaluates the presentation, surgical therapy and recurrence of these three proliferative disorders. We retrospectively reviewed the clinical data from all cases of GCTTS, TGCT, and PVNS from 1985 to 1997. A total of 35 patients were identified: GCTTS (n=8), TGCT (n=1), and PVNS (n=26), there were 18 men and 17 women. The median age was 48 years (range 6-84 years). The most common site of involvement was the knee (15), followed by wrist (7), elbow (4), and hip (4). Seven patients had extra-articular involvement, and 19 were found incidentally at operations for other reasons. Among the 4 patients who developed recurrent disease, 2 had extra-articular disease at the time of their original diagnosis. None died, and none required major amputation. One patient was treated with adjuvant radiotherapy following resection of a recurrence. It is important to distinguish between focal and diffuse forms of synovial involvement. If focal, simple surgical excision is appropriate. If diffuse, complete synovectomy is indicated for disease confined to the joint, and resection of all gross disease is indicated for extra-articular disease. Radical resection with negative margins is not necessary in most instances. In rare aggressive cases, local recurrence may necessitate more extensive resection and radiation therapy. PMID- 10671696 TI - Alterations in the retinoblastoma pathway of cell cycle control in parathyroid tumors. AB - Mutations in proto-oncogenes and tumor suppressor genes have been associated with tumor development and/or progression in many neoplasms. It has been reported that parathyroid tumors have deletions affecting the retinoblastoma gene (RB), and overexpression of cyclin D1 (Cyc D1). The aim of the present study was to evaluate the alterations in the components of the pRB pathway in parathyroid adenomas and parathyroid aggressive tumors, including patterns of expression of pRB, Cyc D1, and p16/INK4A. Paired normal and tumor DNA from 6 parathyroid adenomas and 5 aggressive tumors were analyzed for loss of heterozygosity (LOH) at the RB locus. The expression of pRB, Cyc D1 and p16 was studied in 4 adenomas and 5 aggressive tumors. RB LOH was found in 1 of 6 adenomas, and in 1 of 2 informative aggressive tumors. Immunohistochemical analysis revealed undetectable pRB in 4 of 5 aggressive tumors and presence of pRB in all adenomas. Conversely, Cyc D1 expression was found in 3 of 4 aggressive tumors, but was undetectable in the adenomas. Expression of p16 was identified only in one aggressive tumor. Thus, alterations in the pRB pathway seem to prevail in the aggressive form of parathyroid neoplasms. Our results warrant further investigation of these cell cycle regulators in order to determine their potential role as tumor markers in parathyroid tumors. PMID- 10671697 TI - Allelic imbalance at NBS1 is frequent in both proximal and distal colorectal carcinoma. AB - Nijmegen breakage syndrome (NBS) is a hereditary disorder involving chromosomal instability, cancer risk and radiosensitivity. NBS carriers have an increased risk of cancer, though the significance of mutations in the NBS1 gene in sporadic cancer has not yet been investigated. Because the loss of NBS1 is associated with increased chromosomal re-arrangements, and tumors of the colon are particularly prone to chromosomal anomalies, we have begun to study the NBS1 locus in colorectal cancer (CRC). DNA was isolated from 99 microdissected colorectal tumors, and microsatellite markers flanking the NBS1 locus at 8q21.3 as well as elsewhere on 8q were analyzed. Normal lymphocyte DNA from each patient served to normalize the amplification of each allele, and a reduction of at least 35% in the intensity of one allele was taken as evidence of allelic imbalance (AI). In proximal and distal CRCs we found 25.9 and 36.2% with AI at 8q21.3, respectively. AI in proximal CRC tended not to extend to marker D8S555 at 8q24.1, whereas in distal CRC the region of AI frequently included all the informative markers. AI of 8q21.3 was not associated with any clinical variable. These results suggest that 8q21.3 contains a tumor suppressor gene involved in proximal CRC, possibly NBS1. The large regions of AI make it difficult to determine the importance of AI at the NBS1 locus in distal CRC. PMID- 10671698 TI - High-resolution DNA flow cytometry in oral verrucous carcinoma. AB - The classification of verrucous carcinoma as an entity unto itself or as a variant of well differentiated squamous cell carcinoma is controversial. To contribute new insights into the biological behavior of this rare tumor, we applied DNA flow cytometry to three node-negative verrucous carcinomas of the oral cavity. All tumors expressed a single aneuploid cell population. One of the patients experienced three courses with local recurrence. All secondary tumors retained the initially established aneuploid clone. The development of aneuploidy is thus a cytogenetic event common to both verrucous and squamous carcinoma of the oral cavity. PMID- 10671699 TI - Homeopathic treatment of mild traumatic brain injury: A randomized, double-blind, placebo-controlled clinical trial. AB - BACKGROUND: Mild traumatic brain injury (MTBI) affects 750,000 persons in the United States annually. Five to fifteen percent have persistent dysfunction and disability. No effective, standard pharmacological treatment exists specifically for this problem. We designed a pilot research project to study the clinical effectiveness of homeopathic medicine in the treatment of persistent MTBI. METHOD: A randomized, double-blind, placebo-controlled trial of 60 patients, with a four-month follow-up (N = 50), was conducted at Spaulding Rehabilitation Hospital (SRH). Patients with persistent MTBI (mean 2.93 years since injury, SD 3.1) were randomly assigned to receive a homeopathic medicine or placebo. The primary outcome measure was the subject-rated SRH-MBTI Functional Assessment, composed of three subtests: a Difficulty with Situations Scale (DSS), a Symptom Rating Scale (SRS), and a Participation in Daily Activities Scale (PDAS). The SRH Cognitive-Linguistic Test Battery was used as the secondary measure. RESULTS: Analysis of covariance demonstrated that the homeopathic treatment was the only significant or near-significant predictor of improvement on DSS subtests (P =.009; 95% CI -.895 to -.15), SRS (P =.058; 95% CI -.548 to.01) and the Ten Most Common Symptoms of MTBI (P =.027; 95% CI -.766 to -.048). These results indicate a significant improvement from the homeopathic treatment versus the control and translate into clinically significant outcomes. CONCLUSIONS: This study suggests that homeopathy may have a role in treating persistent MTBI. Our findings require large-scale, independent replication. PMID- 10671700 TI - Measuring psychosocial recovery after traumatic brain injury: psychometric properties of a new scale. AB - OBJECTIVES: To determine the psychometric properties of the Sydney Psychosocial Reintegration Scale (SPRS), an instrument developed to quantify disability and handicap in persons with traumatic brain injury (TBI). DESIGN: Descriptive correlational study. SETTING: Brain Injury Rehabilitation Unit and Brain Injury Outpatient Clinic. PARTICIPANTS: Two samples, a "subacute" group (n = 20) and a "long-term" group (n = 40), were studied to examine responsiveness (subacute group), reliability, and validity (long-term group) of the SPRS. MAIN OUTCOME MEASURE: The SPRS is a 12-item questionnaire measuring three domains of everyday living commonly disrupted after severe TBI: occupational activities, interpersonal relationships, and independent living skills. PROCEDURE: Patients in the subacute group were rated with the SPRS by a clinician at admission to the rehabilitation unit and again three months later or at discharge from the unit (whichever occurred first). For individuals from the long-term group attending the outpatient clinic, a close relative was interviewed with the SPRS and other validating measures. The SPRS was readministered one month later. RESULTS: Internal consistency of the SPRS was high (alpha coefficient = .90), as was agreement between raters and stability over a one-month period (r(i) = .95 and .90, respectively). Reliability and stability coefficients for the three domains of the scale were also high, ranging from.86 to.94 for reliability and.77 to.93 for stability. Preliminary evidence for construct validity was established with a number of standard instruments, with evidence of both convergent and discriminant construct validity from the Sickness Impact Profile (SIP). The SPRS was sensitive to group differences on the Glasgow Outcome Scale (GOS) and to changes occurring during the period of active recovery. CONCLUSIONS: The results suggest that the SPRS has sound psychometric properties, being a reliable, stable, sensitive, and valid instrument. It is potentially useful in both clinical and research settings. PMID- 10671701 TI - Functional outcome of individuals with traumatic brain injury and lower extremity deep venous thrombosis. AB - OBJECTIVE: To determine the impact of acute lower extremity (LE) deep venous thrombosis (DVT) on functional outcome after traumatic brain injury (TBI). SETTING: Tertiary university medical center rehabilitation unit. SUBJECTS: Ninety two TBI rehabilitation patients (46 patients with DVT and 46 patients without DVT). Forty-six TBI patients with a diagnosis of LE DVT were 1:1 matched with non DVT TBI patients. Matching criteria included: primary diagnosis of TBI, admission Functional Independence Measure (FIM), Glasgow Coma Scale (GCS), and age. OUTCOME MEASURES: FIM (admission, discharge, change, and efficiency), FIM subscores (activities of daily living [ADL], mobility, cognition), length of stay ([LOS] acute and rehabilitation), and discharge living disposition. DESIGN: Cohort study utilizing prospectively collected data. DVT diagnoses were made upon rehabilitation admission using color flow duplex Doppler ultrasonography. Descriptive statistics were run on demographic variables. Analyses of variance (ANOVAs) were performed on the sample with regard to outcome measures, including FIM scores, FIM subscores, and LOS (acute and rehabilitation). RESULTS: No significant between-group differences were found concerning LOS, rehabilitation costs, FIM total, or FIM subgroup scores. Chi-squared analyses revealed significant differences between groups with regard to discharge living disposition (chi(2) = 4.7, P <.03). CONCLUSION: Lower extremity DVT does not appear to interfere with functional outcome after TBI. The data suggest that this patient population is appropriate for admission or continued participation in acute inpatient rehabilitation, despite the presence of LE DVT. PMID- 10671702 TI - Sex offending as a psychosocial sequela of traumatic brain injury. AB - OBJECTIVE: To describe the nature and extent of sexual offending after traumatic brain injury (TBI). DESIGN: Retrospective file review. SETTING: A brain injury unit providing inpatient and outpatient rehabilitation services. PARTICIPANTS: A review of five years of admissions to the Brain Injury Rehabilitation Unit (N = 477) identified a sample of 29 males who committed 128 incidents of sex offending. MAIN OUTCOME MEASURES: A protocol to record data on demographic, injury, radiological, and psychosocial variables and offending behaviors. RESULTS: Of the total population of 445 clients with TBI, 6.5% (n = 29) were identified as having committed some form of sexual offense. Alcohol was a factor in only three (2.3%) of the incidents, and only two clients had a preinjury history of sexual offending. The most common offenses were the "touching" offenses, followed by exhibitionism and overt sexual aggression. Staff members were the most common targets of the offenses, followed by members of the general public, other people with TBI, and family members. CONCLUSIONS: Sex offending is a significant clinical problem among a small minority of men after TBI. The absence of alcohol and preinjury histories of sexual offending suggest that the brain injury and contingent sequelae were a significant etiological factor underlying the offenses. A number of implications for the clinical management of clients with sexually aberrant behaviors is identified and discussed. PMID- 10671703 TI - Neuropsychological significance of anosmia following traumatic brain injury. AB - OBJECTIVES: To investigate the incidence of anosmia following traumatic brain injury (TBI) using a standardized instrument and to test hypotheses that post-TBI anosmics perform significantly more poorly than do post-TBI normosmics on measures of executive skills and functional outcome. DESIGN: Prospective quasi experimental between-groups design. PARTICIPANTS: Sixty-eight adults diagnosed with TBI. SETTING: Brain injury rehabilitation program based at a Midwestern medical center. MAIN OUTCOME MEASURES: University of Pennsylvania Smell Identification Test (UPSIT), selected neuropsychological measures of executive skills, the Disability Rating Scale (DRS), and the Community Integration Questionnaire (CIQ). RESULTS: Forty-four subjects (65%) demonstrated impaired olfaction; only 13 (30%) acknowledged smell dysfunction. Anosmic and normosmic groups did not differ in demographics, IQ, chronicity, or admission Glasgow Coma Scale (GCS). Anosmics had longer coma (P =. 01), more severe deficits in complex attention (Trailmaking Test, Part B, P =.01), new learning/memory (California Verbal Learning Test Trial V [CVLT-V], P =.001), and problem solving (Wisconsin Card Sorting Test [WCST], P =.001), leading to greater functional impairment (Disability Rating Scale [DRS], P =.003). No differences emerged on the CIQ. CONCLUSIONS: Anosmia is a common sequela of TBI, although only a minority of patients are aware of this deficit. Further, anosmics demonstrated greater impairment in a variety of frontal-lobe mediated executive functions, as well as greater functional disability. PMID- 10671704 TI - Interaction of posttraumatic stress disorder and chronic pain following traumatic brain injury. AB - OBJECTIVE: To investigate the association between posttraumatic stress disorder (PTSD) and chronic pain in patients who had sustained a severe traumatic brain injury (TBI). DESIGN: Correlational relationships between pain variables and PTSD measures were examined in a cohort study. SETTING: An adult tertiary care center brain injury clinic. PATIENTS: Ninety-six persons with severe TBI. OUTCOME MEASURES: The Posttraumatic Stress Disorder Interview (PTSD-I), a modified McGill Pain Questionnaire, the Beck Depression Inventory (BDI), the General Health Questionnaire (GHQ), the Community Integration Questionnaire (CIQ), the Satisfaction with Life Scale (SWL), and the Coping Style Questionnaire (CSQ). RESULTS: More persons with chronic pain reported PTSD than did those without pain. The relationship between pain severity and depression, functional adjustment, and satisfaction with life was mediated by severity of PTSD. Pain severity was significantly associated with an avoidant coping style. CONCLUSIONS: Effective rehabilitation of persons with chronic pain following severe TBI should recognize the role of posttraumatic stress in the maintenance of dysfunctional reactions. Specific interventions that address adaptive coping mechanisms to reduce PTSD may enhance rehabilitation for persons with TBI who suffer chronic pain. PMID- 10671705 TI - Comparison of remedial and compensatory interventions for adults with acquired brain injuries. AB - OBJECTIVE: To examine the effects of a compensatory intervention versus a remedial intervention for deficits in visual processing of adults with acquired brain injuries (ABI). SETTING: A cognitive rehabilitation program at a large comprehensive rehabilitation hospital in the New York City metropolitan area. PATIENTS: Thirty adults with ABI were matched according to severity of injury, gender, age, and time post-injury, and randomly assigned to the remedial or compensatory group. INTERVENTIONS: The remedial intervention consisted of four 45 minute sessions (once weekly) of participation in computer tasks without instruction in compensatory strategies. The compensatory intervention consisted of four 45-minute sessions of instruction in the use of three internal compensatory strategies, including verbalization, chunking, and pacing. MAIN OUTCOME MEASURES: Pretest/posttest measures included three functional computer tasks. Weekly measures included a computerized version of the Paced Auditory Serial Addition Task (PASAT) and two computerized matching tasks. RESULTS: Both groups exhibited statistically significant improvement of comparable degree on posttests and weekly measures. Further analysis revealed that 80% of both groups used compensatory strategies, regardless of intervention method. Those who used strategies demonstrated better performance than those who did not. CONCLUSION: The ability to use internal compensatory strategies may be a significant confound in research examining the effects of the various cognitive rehabilitation intervention methods. PMID- 10671706 TI - Traumatic brain injury in the United States: A public health perspective. AB - Traumatic brain injury (TBI) is a leading cause of death and disability among persons in the United States. Each year, an estimated 1.5 million Americans sustain a TBI. As a result of these injuries, 50,000 people die, 230,000 people are hospitalized and survive, and an estimated 80,000-90,000 people experience the onset of long-term disability. Rates of TBI-related hospitalization have declined nearly 50% since 1980, a phenomenon that may be attributed, in part, to successes in injury prevention and also to changes in hospital admission practices that shift the care of persons with less severe TBI from inpatient to outpatient settings. The magnitude of TBI in the United States requires public health measures to prevent these injuries and to improve their consequences. State surveillance systems can provide reliable data on injury causes and risk factors, identify trends in TBI incidence, enable the development of cause specific prevention strategies focused on populations at greatest risk, and monitor the effectiveness of such programs. State follow-up registries, built on surveillance systems, can provide more information regarding the frequency and nature of disabilities associated with TBI. This information can help states and communities to design, implement, and evaluate cost-effective programs for people living with TBI and for their families, addressing acute care, rehabilitation, and vocational, school, and community support. PMID- 10671707 TI - Mapping variation in brain structure and function: implications for rehabilitation. AB - The recognition of structural and functional variability of the human brain promoted the development of a system to organize and analyze the rapidly growing body of knowledge acquired among diverse disciplines. The Human Brain Project (HBP) was initiated and has evolved as a means of establishing an information infrastructure through tools related to the emerging science of neuroinformatics. This article will briefly describe the implications that such an endeavor has for the field of traumatic brain injury (TBI) rehabilitation. PMID- 10671708 TI - [Q.A.L.Y. index and assessment of cost efficiency of the cochlear implant in acquired profound deafness]. AB - Cost utility analysis is a method of cost-effectiveness analysis which provides results in terms of cost per quality-adjusted life-year (Q.A.L.Y.). This method is progressively largely used for medical technology assessment. It permits cost effectiveness comparisons between medical interventions. The cost per Q.A.L.Y. of cochlear implant was determined in 30 adult patients suffering from acquired profound deafness. This study indicates that this technology provides significant improvements and is quite cost-effective. However, as far as profound deafness is concerned, the reliability of Q.A.L.Y. needs to be improved. PMID- 10671709 TI - [Signs and symptoms, etiologies and clinical course of parosmia + in a series of 84 patients]. AB - Eighty-four patients (72% females and 28% males) consulted between January 1995 and January 1998 for olfactory disorders with parosmia (erroneous olfactory response to stimuli). Parosmia occurred immediately after or during the course of acute rhinitis (n=70, 83%), head trauma (n=7, 9%), naso-sinus polyposis (n=5, 6%), chronic rhinitis (n=1) or frontal tumor (n=1). Quantitative and qualitative olfactory disorders were analyzed and products producing the parosmia were identified. The only cases where parosmia regressed concerned patients who developed parosmia after acute rhinitis (n=28 cases, 33%). The prognosis of parosmia appeared to be better when it was a secondary phenomenon: i) the percentage of improvements was higher though not significant (41% versus 26.7% compared with primary parosmia), ii) delay to improvement was shorter (8.4 +/- 2.1 months versus 14.5 +/- 4.4 months for primary parosmia, p<10(-4) ), iii) there were no cases of persistent parosmia where quantitative disorders improved (compared with 7 cases of persistent primary parosmia, p<0.05). In addition, the prognosis of associated quantitative disorders was the same for both primary and secondary parosmia. Only the delay to improvement appeared to be shorter in case of secondary parosomia (though the difference was not significant). Products which produced the parosmic perception were identified by nearly all the patients (85%). The most frequently cited products were coffee, perfume, certain fruits (melon, banana, citric fruits), tobacco or chocolate. All these products contain tannic acid, a water-soluble polyphenol with many biological properties (influence feeding habits and metabolism in the rate, antioxidant and antimutagenic properties). Thus acid tannic could induce parosmic perception due to its antioxidant properties susceptible of integrating the P-450 cytochromes of the mucosal cells or olfactory neuroepithelium supporting cells. PMID- 10671710 TI - [Orbital complications of sinusitis in adults]. AB - Orbital complications of sinusitis are rare in adults but delayed diagnosis is vision and life threatening. We report our experience in 6 patients to present clinical history, bacteriology and discuss the modality of treatment. There were 4 young men and 2 women, aged from 16 to 79 years old. Only one patient had an immunocompromised underlying condition (HIV infection). Four patients had preseptal abscesses and three post septal cellulitis or abscess (one patient had preseptal abscess and post septal abscess and hematoma). Two patients had a complete unilateral loss of light perception. Pathogens encountered were Streptococcus species: 4, strict anaerobes: 1, Pseudomonas aeruginosa: 1 (patient with AIDS). Patients recovered from infection with antibiotics in 6 and surgery in 5 but sequellar blindness occurred in 2 patients. Our experience emphasizes the necessity of antibiotic treatment in bacterial sinusitis and importance of early diagnosis and appropriate management of complications. PMID- 10671711 TI - [Swallowing disorders in unilateral recurrent laryngeal nerve paralysis]. AB - Swallowing difficulties in isolated recurrent laryngeal nerve paralysis are rarely reported in the literature, although it is frequently mentioned in lesions of both recurrent laryngeal and superior laryngeal nerves. The aim of this prospective study was to evaluate the incidence of aspiration in patients with unilateral recurrent laryngeal nerve paralysis after head and neck or thoracic surgery. Eighteen patients were included and evaluated within the first week and eleven two months postoperatively. Position, tone and tension of the vocal fold as well as assessment of the glottic axis, arytenoid position and mobility, laryngeal sensibility, status of the pyriform sinus and salivary stasis were studied. Swallowing evaluation was performed using flexible fiberoptic videolaryngoscopy during dry swallowing, thick cream and methylene blue liquid swallowing. Five patients had symptomatic aspiration, one silent aspiration and twelve patients had no aspiration. The type of regimen feeding used was classified as a normal, mixed, or blended diet. Added specific treatment performed was also described. We conclude that aspiration may occur in unilateral recurrent laryngeal nerve paralysis and have to be systematically evaluated after pneumomectomy, because adequate treatment can be proposed. PMID- 10671712 TI - [Replacement of tracheo-esophageal Provox prosthesis]. AB - OBJECTIVES: To compare anesthesic techniques used between 1992 and 1997 at Laennec Hospital for replacement by tracheo-esophageal Provox prosthesis: local and general anesthesia. Theoretical financial cost for replacement was estimated according to anaesthetic techniques. PATIENTS AND METHODS: Provox in situ lifetime was calculated in 58 patients who underwent 115 and 49 replacements under general and local anaesthesia respectively. Age, sex, surgical and radiotherapy backgrounds, complications and anaesthetic techniques were studied as potential factors correlated with Provox in situ lifetime. Theoretical financial cost for replacement was estimated according to anaesthetic techniques. RESULTS: In 1992, 12% of Provox prosthesis were inserted under local anaesthesia and 54% in 1997. Provox in situ lifetime was either not influenced by anaesthetic techniques or other factors under analysis. The theoretical financial cost was estimated at 14, 341 FFrs and 6,048 FFrs for replacement under general and local anaesthesia respectively. CONCLUSION: Due to increased control of health care costs, we advocated local anaesthesia for Provox prosthesis replacement if control endoscopy is not required. PMID- 10671713 TI - [Thyroid cancer treatment]. AB - INTRODUCTION: Thyroid diseases are common although cancer is rare. There are some controversial issues concerning the extent of surgical treatment of thyroid cancer. MATERIALS AND METHODS: We have studied 614 cases of thyroidectomy inducing 82 of mostly papillary and follicular thyroid cancers. RESULTS AND DISCUSSION: We observed that differentiated thyroid cancer has a predilection for the right lobe and one third of papillary tumors are multifocal. We have seen that fine needle aspiration cytology is the most useful preoperative study. Intraoperative frozen biopsy has a good specificity but sensitivity is low in our series, specially for follicular neoplasms. The treatment in our series consisted in total thyroidectomy and, in differentiated cancers, postoperative I-131. Survival is very good for differentiated cancers. Prognosis is poor for anaplastic carcinoma in the short term. PMID- 10671714 TI - [Interest of free flaps for cranial base reconstruction]. AB - We describe our experience in treating 7 patients who underwent skull base reconstruction with free flap (6 latissimus dorsi, 1 rectus abdominis) between October 1996 and November 1998. Four patients underwent temporal bone resection with auricular resection, 2 patients underwent anterior and middle cranial fossa resection, 1 patient underwent frontotemporal resection. There have been no failures of the free flaps and one cerebrospinal fluid leak. We advocate free flap reconstruction after temporal bone resection with auricular resection, and after anterior or middle cranial fossa resection when local flap options are not available or with complex dead space. PMID- 10671715 TI - [Endoscopic resection of the frontal sinus floor]. PMID- 10671716 TI - [ [In Process Citation] PMID- 10671717 TI - [ [In Process Citation] PMID- 10671718 TI - [Cardiac MR: morphological and functional imaging]. PMID- 10671719 TI - [Abdominal complications of Crohn's disease: CT features]. AB - Crohn's disease is characterized by transmural inflammation and chronic disorder of the gastrointestinal tract. Abdominal complications of Crohn's disease are frequent and quite variable and their diagnosis is commonly performed with CT. The purpose of this article is to review the CT features of the abdominal complications of Crohn's disease, including ileocolitis, abscess, phlegmon, fistula, bowel obstruction, portal vein gas, colonic distention, as well as urinary, hepatobiliary and pancreatic complications. PMID- 10671720 TI - [A joint continuing medical education program for practitioners and radiologists: a positive preliminary experience]. AB - PURPOSE: A common reflective continuing medical education (CME) approach has been promoted by the French National CME Association (UNAFORMEC) and French Radiologic Organizations to improve imaging prescriptions performed by general practitioners (GP) as well as their relationships with radiologists to fit the best decision resources for an optimal patient care. This new practice based CME is a multidisciplinary learning model. MATERIALS AND METHODS: A learning model that could satisfy GP and favor improved communication between them and radiologists was elaborated. Two organization groups worked simultaneously during one year before the first CME session could be achieved. This kind of CME involves a small group (30 participants, 3/4 of general practitioners and 1/4 of radiologists), and includes 3 sessions; on the basis of fictive medical situations, the first one addresses to the formulation of a pertinent imaging prescription, including data that are useful to the radiologist; the second one involves a critical point of view of the imaging interpretation by the radiologist; the last one emphasizes the communication between the patient, the GP and the radiologist. RESULTS: At the present time two CMEs have been performed. Immediate participant appreciation was enthusiastic and only some minor revisions were needed. Further evaluation upon changes of practices has now to be made. CONCLUSION: This CME deserves to provide not only a new practice learning model, but also a humanistic perspective. PMID- 10671721 TI - [Value of brain MR imaging in infants with a severe idiopathic apparent life threatening event]. AB - OBJECTIVE: Prognostic value of a magnetic resonance imaging (MRI) scoring system in infants with a severe apparent life threatening event (ALTE). METHODS: Ten infants with an ALTE (aged between 6 and 31 weeks) were clinically graded according to the PRISM score and evaluated with EEG, evoked potentials and MRI. The 18 MRIs obtained were distributed in 3 classes according to the delay after which they were obtained; class A (n=5): within the first 48 hours after the event, class B (n=7): between day 3 and 8 and class C (n=6): between day 9 and 50. The 18 MRIs were evaluated retrospectively using a scoring system based on 3 categories of lesions: edema, basal ganglia injury and watershed injuries. Five infants died between day 2 and day 15 after the event. The five surviving infants had follow up neurodevelopmental testing after 38 to 77 months. RESULTS: There was no correlation between the 5 MRIs of class A and the neurological outcome. For the MRIs of class B and C, the scoring system can be of great value when combined with the scores of EEG, EP and PRISM. CONCLUSIONS: The scoring system for MRI performed within 48 hours after the event is falsely reassuring. MRI can be helpful as early as 3 days after the event when combined with the score of the electrophysiological investigations and the PRISM. PMID- 10671723 TI - [Carney's triad: update and report of one case]. AB - The Carney's syndrome associates three different tumors on the same subject, a young woman generally: a gastric leiomyosarcoma, a pulmonary chondroma and a non adrenal paraganglioma. The authors report a new case of that unfrequent syndrome of unexplained pathogeny. PMID- 10671722 TI - [Lumbar intraspinal synovial cysts: imaging and treatment by percutaneous injection. Report of thirteen cases]. AB - PURPOSE: The CT and MR imaging findings in 13patients with lumbar intra spinal synovial cysts were retrospectively analysed and the results of facet corticosteroid injection were evaluated. PATIENTS AND METHODS: Over a 7 year period, 13patients with radicular pain were identified as having lumbar intra spinal synovial cysts. They ranged from 42 to 87 years of age. All patients were evaluated by CT without contrast material and underwent facet arthrography associated with corticosteroid injection and CT arthrography. MR imaging was performed in all patients either before or after percutaneous treatment. CT scans and MR images were reviewed and patient outcome was evaluated at 1and 6month followup. RESULTS: CT scan revealed a cystic structure adjacent to a degenerated facet joint in 9 patients (69% sensitivity). MRI showed more accurately the cyst on T2 weighted and/or axial images. Complete or good relief of radicular pain and functional restrictions were achieved in 9 patients (69%) at 1 month follow up, still to be found in 6patients (46%) at 6months. CONCLUSION: In patients with radiculopathy and facet degenerative changes, intra spinal synovial cysts must be looked for. Facet corticosteroid injection is a useful alternative to surgical removal. PMID- 10671724 TI - [Atypical sonographic findings of bacterial colitis]. AB - We report three pediatric cases of infectious colitis that were misinterpreted on US examination as Crohn's disease. These colitis were limited to the left colon and presented with transmural hypoechoic thickening of the wall and homogenous hyperechoic appearance of the surrounding fat. PMID- 10671725 TI - [Angiosarcoma of the breast. Radiological aspects. About one case]. AB - Angiosarcoma of the breast in an uncommon entity. The authors report a well documented case of angiosarcoma. The patient, a 23 years old woman, had a voluminous mass of the left breast without inflammation. The evolution was rapidly fatal. Physical examination revealed a large painful breast mass with purplish discoloration cutaneous area in front of the lesion. The palpation revealed a thrill. The mammography had shown an area of increased density in the left breast. A complementary ultrasound examination detected a well circumscribed voluminous mass with hypoechogenic heterogenous echostructure. The Doppler examination detected a venous blood flow in the tumor. This finding was compatible with vascular tumor as angioma or angiosarcoma. The angioscanner showed a peripheral vascular enhancement and a centripetal diffusion of the contrast product. This vascular kinetic is seen commontly in the angioma. Magnetic resonance imaging in the T1 and T2 relaxation times detected an intermediate signal mass with bleeding areas. The patient had had a mastectomy and the histopathologic examination confirmed the diagnostic of angiosarcoma. The radiological aspacts of angiosarcoma of the breast are discussed depending on the literature data. PMID- 10671726 TI - [Cortical septic osteitis: two cases]. AB - Cortical septic osteitis is defined as a predominant or exclusive infection of the cortex which may lead to diagnosis pitfalls. With the two cases presented here, principal radiological features allowing a correct diagnosis are recalled. PMID- 10671727 TI - [What is it? Left-sided appendicitis]. PMID- 10671728 TI - [Percutaneous osteosynthesis of pelvic fractures with CT control]. AB - Pelvis fractures, most often multiple, are frequently unstable. Orthopedic treatment is hardly bearable (traction in bed sometimes up to 45 days), the open reduction and internal fixation (ORIF) is heavy. Percutaneous fluoroscopy guided fixation lacks precision in depth. Percutaneous screw fixation with CT scan control answers these drawbacks and represents a quick solution, with few hazard when performed by a trained team and allows a very early resumption of standing. PMID- 10671729 TI - [Resection of osteoid osteoma]. PMID- 10671749 TI - Embryonic stem cells and nuclear transfer strategies. Present state and future prospects. PMID- 10671750 TI - A role for epithelial-mesenchymal interactions in tail growth/morphogenesis and chondrogenesis in embryonic mice. AB - Neurulation involves development from primary germ layers before any differentiation of embryonic mesenchyme. Subsequently, secondary organogenesis is via epithelial-mesenchymal interaction. It is unclear whether formation of the caudal body axis and tail bud in vertebrate embryos is by temporal and causal extension of primary neurulation, by secondary neurulation, or by secondary induction (epithelial-mesenchymal interactions) as seen in organogenesis of the limb buds, kidneys, heart and other embryonic regions. Reports of a ventral ectodermal ridge (VER) associated with tail bud development in rodent embryos imply that tail bud development may share features with limb bud development, in which the apical ectodermal ridge (AER) directs limb bud outgrowth and skeletal patterning. Organ culture or grafting to the chorioallantoic membranes of host chick embryos, of tail bud mesenchyme with or without tail epithelium, demonstrates that both survival and growth of tail mesenchyme depend on the presence of tail epithelium. Initiation of chondrogenesis of tail mesenchyme was similarly dependent on tail epithelium until 10.5 days of gestation, which is when the VER is at its maximal extent. Initiation of myogenesis was independent of the presence of tail epithelium. These results are discussed in relation to the similarity of tail bud to limb bud developed, and to the different mechanisms employed in differentiation of the cranial and caudal ends of vertebrate embryos. Secondary induction of the caudal body region is argued to be fundamental in vertebrate embryogenesis. PMID- 10671751 TI - Leptin expression during the differentiation of subcutaneous adipose cells of human embryos in situ. AB - In this study, the immunocytochemical expression of leptin in the developing subcutaneous tissue of human embryos at 6-10 weeks of gestation was investigated using the avidin-biotin peroxidase method. Immunocytochemical staining for leptin was observed in the cytoplasm of the differentiating preadipose cells. The other cells present in the embryonal subcutis (mesenchymal cells differentiating into fibroblasts, fibrocytes, endothelial cells) were leptin-negative. The developing skin epithelium lying above it and the blood cells in the capillaries also did not express leptin. The results suggest that the leptin is produced by the developing fat cells from the beginning of lipidogenesis and differentiation. It possibly acts as a hormonal factor regulating the embryonal growth, development and body fat storage. PMID- 10671752 TI - Intraepithelial leucocytes in the bovine uterine tube. AB - The epithelium of the uterine tube consists of ciliated cells and secretory cells. Basal cells are a third cell type observed in tubal epithelium and they are located principally in the basal part of the epithelium. The objectives of this study were to characterize these basal cells in normal and superovulated heifers and to determine whether they participate in the replacement of the ciliated and secretory cell populations. All heifers received cloprostenol (PG) to induce oestrus (day 0). Superovulated heifers received 24 mg pFSH at doses of 4.5, 3.5, 2.5 and 1.5 mg given twice daily. Control and superovulated heifers were slaughtered on days 1, 3, 5 and 7 of the oestrous cycle. Another group of normal cycling heifers was slaughtered on days 2-3 and 11-13 of the oestrous cycle and used for immunocytochemistry. Samples from ampulla, pre-isthmus and isthmus of the uterine tube were collected and processed for light and transmission electron microscopy. Quantitative examination by light microscopy showed that there was a significant difference in the number of basal cells between the regions of the heifers' uterine tube. On the basis of ultrastructure two populations of basal cells were observed. One (type I) had a nucleus with much condensed heterochromatin and very sparse cytoplasmic organelles. The second cell (type II) had a nucleus with heterochromatin typically clumped around the nuclear envelope. Its cytoplasm contained many organelles including a number of lysosomes. The ultrastructural features of these cells were similar in all regions and at all days of the oestrous cycle examined. Immunocytochemistry revealed that type I basal cells were lymphocytes and type II basal cells were macrophages. PMID- 10671753 TI - Immunocytochemical and immunoelectron-microscopic study of somatotrophs in ICR and nonobese diabetic mice. AB - Somatotrophs (GH cells) were classified immunoelectron microscopically into three types mainly on the basis of the size of secretory granules in the mouse pituitary of ICR strain. Type I cells contained large secretory granules. Type II cells contained both large secretory granules and small secretory granules. Type III cells contained small secretory granules. All three types of GH cells were found from the neonatal ages to adult. The relative proportion of three types did not change with age, and no sex differences in the relative proportion of the cell types were detected. Type I cells predominate in all age groups observed. In 60-day-old male mice percentages of each type were as follows: type I 93.7 +/- 0.1%, type II 5.4 +/- 0.8%, and type III 0.9.0 +/- 0.4% (n = 5), and in 60-day old female mice type I 95.2 +/- 0.1%, type II 2.7 +/- 0.5%, and type III 2.1 +/- 0.9% (n = 5). The maximum diameters of the large secretory granules increased from 7 to 60 days of age. The small secretory granules similarly increased in size in female mice, but those in male mice did not change. In the diabetic female mice of the nonobese diabetic (NOD) strains, GH cells in diabetic mice became smaller, and the number and size of secretory granules decreased, indicating diminished GH secretion. However, the relative proportion of each subtype of GH cells did not differ irrespective of the occurrence of diabetes. PMID- 10671754 TI - Distribution of interglobular dentine in human tooth roots. AB - The present study was designed to examine the distribution of interglobular dentine in human tooth roots. The material comprised 17 teeth, of which 3 were premolars extracted for orthodontic reasons from children 10-12 years of age and the other teeth (4 incisors, 3 canines and 7 molars) were extracted for periodontitis from individuals aged 32-63 years. All teeth were free of caries and cervical dentine defects. Ground sections of the teeth cut longitudinally were stained with basic fuchsin and observed by fluorescence and confocal microscopy as well as transmitted light microscopy. Basic fuchsin stained the dentinal tubules, interglobular dentine and the granular layer of Tomes. These structures appeared intense blue to faint violet with transmitted light microscopy, whereas their staining displayed intense fluorescence with fluorescence microscopy. Therefore, the interglobular dentine could be detected more sensitively with fluorescence and confocal microscopy than with transmitted light microscopy. Typical interglobular dentine was present in coronal dentine in most of the teeth. In the radicular dentin, position and size of the interglobular dentine was different among the teeth examined. Most of the teeth had the interglobular dentine in the cervical part of the roots (type A). Two premolars displayed the interglobular dentine in the coronal half of the root (type B). The types A and B contained large interglobular areas. A small amount of interglobular dentine was restricted to the apical half of the roots of two canines and one molar (type C). In contrast to types A and B which were seen at both labial or buccal and lingual sides of roots, the interglobular dentine of type C was seen only at one side, labial or lingual. Some of the tooth roots did not show any interglobular dentine (type D). Most of the incisors, canines and premolar were types A, B, and C, respectively, and the molars were mixed types A, C, and D. These results suggest that the factors affecting dentinogenesis during root formation are unique for each tooth. PMID- 10671755 TI - Effects of muscle contraction on the load-strain properties of frog aponeurosis and tendon. AB - The mechanical properties of the frog semitendinosus (ST) tendon and aponeurosis were measured during passive tensile loading to a force equal to ST maximum tetanic tension and during active isometric muscle contraction. During active contraction, both the tendon and aponeurosis regions initially strained at rates exceeding 400%/s while near the end of the muscle contraction, strain rates were nearly zero. At this point, the strain in the tendon region was equal to that observed during slow passive loading to the same tension level. However, for the aponeurosis, even near the zero strain rate, strain at the end of the active contraction was significantly below that observed during slow passive loading (p < 0. 001). Specifically, when aponeurosis strain rate was almost zero, aponeurosis strain was 13.8 +/- 3% (means +/- SEM, n = 10), which was significantly below that measured during passive loading (23.7 +/- 5%) suggesting that active contraction actually altered aponeurosis material properties. These data demonstrate that, while the tendon and aponeurosis regions have different passive biomechanical properties and both demonstrate viscosity typical of other connective tissues, the aponeurosis region of the frog ST actually changed its intrinsic properties during muscle contraction. Thus, extrapolation of biomechanical data obtained at nonphysiological strain rates or under conditions where the muscle-tendon junction has been interrupted should be made with caution. PMID- 10671756 TI - Evaluation of neutropenic fever: value of serum and plasma parameters in clinical practice. AB - Treatment-related mortality due to infectious complications following potentially curable aggressive chemotherapy remains a major clinical problem. However, the diagnosis of neutropenic infections is difficult. Although it is common practice to institute empirical broad-spectrum antibiotics in neutropenic fever, liberal use of antibiotics may contribute to increasing resistance and superinfection such as systemic mycosis. Clinicians are searching for a highly specific and sensitive marker indicating early infection. Serum concentrations of several acute-phase proteins (C-reactive protein, serum amyloid A), proinflammatory cytokines (TNFalpha, IL-1, IFNgamma, IL-6, IL-8), soluble adhesion molecules (soluble E-selectin, vascular cell adhesion molecule 1, intercellular adhesion molecule 1) and more recently procalcitonin have been investigated as to whether these may contribute to identifying infections as the cause of neutropenic fever. Unfortunately, at present, based on the small and inconsistent amount of data available from the literature one is tempted to conclude that the predictive values of all these parameters are too low to influence the clinically based initial treatment decisions in patients with neutropenic fever. PMID- 10671757 TI - Flucytosine: correlation between toxicity and pharmacokinetic parameters. AB - Flucytosine (5-fluorocytosine, 5-FC) is a systemic antimycotic drug the major toxicities of which are bone marrow depression and hepatotoxicity. The purpose of this observational and retrospective study was to assess a possible relationship between toxicity and 5-FC pharmacokinetics within a group of 53 intensive care unit patients. The presented results reveal that thrombocytopenia is associated with a decreased 5-FC clearance and that the thrombocyte nadir is linearly related to the 5-FC clearance. Furthermore, patients experiencing 5-FC levels exceeding 100 mg 5-FC/l were found to be at a higher risk of developing thrombocytopenia and hepatotoxicity as compared to those not exceeding this level. PMID- 10671758 TI - In vitro and in vivo antibacterial activities of biapenem in the fields of obstetrics and gynecology. AB - Biapenem is a new injectable carbapenem antibiotic which has favorable pharmacokinetic properties, and is stable to hydrolysis by dehydropeptidase I. Biapenem inhibited more than 90% of clinical isolates of Streptococcus agalactiae, Escherichia coli, Peptostreptococcus magnus, Bacteroides fragilis and Prevotella bivia at the concentration of 3.13 mg/l. The MIC(90) of biapenem against Pseudomonas aeruginosa was lower than that of panipenem, equivalent to that of imipenem, and greater than that of meropenem. The in vivo efficacy of biapenem was evaluated using the experimental infection model of uterine endometritis. The accumulation of neutrophils in the uterus in the biapenem- treated group was less marked than in the nontreated group, as well as bacteriological response. These results suggest that the new antimicrobial agent biapenem might be useful for the treatment of polymicrobial infections in the fields of obstetrics and gynecology. PMID- 10671759 TI - Fluconazole inhibits pseudohyphal growth in Saccharomyces cerevisiae. AB - The ergosterol-depleting antifungal fluconazole, when given at concentrations not affecting growth, inhibited pseudohyphal growth in Saccharomyces cerevisiae, a genetically tractable fungus closely related to the human pathogen Candida albicans. These results suggest that S. cerevisiae could serve as a useful genetic system to study the morphogenetic effects induced by azoles in pathogenic yeast. PMID- 10671760 TI - In vitro susceptibilities of enterococcal blood culture isolates from the Hamburg area to ten antibiotics. AB - Treatment of enterococcal infections is often difficult because of intrinsic and acquired resistance to a variety of antimicrobial agents. Between January 1993 and May 1997, enterococci were isolated from blood cultures of 117 patients at the Institute of Medical Microbiology and Immunology, University Hospital Eppendorf, Hamburg, Germany. Eightynine (76%) isolates were phenotypically identified as Enterococcus faecalis, and 24 (21%) as Enterococcus faecium. All E. faecalis isolates, but only 17% of the E. faecium isolates were susceptible to ampicillin. Two E. faecium isolates (8%) but no E. faecalis were vancomycin resistant (vanA genotype). Quinupristin/dalfopristin shows a high degree of susceptiblity of E. faecium (79%) and may be suitable for the therapy of infections caused by glycopeptide-resistant E. faecium strains. PMID- 10671761 TI - Effect of Varidase (streptokinase) on biofilm formed by Staphylococcus aureus. AB - Staphylococcus aureus forms a fibrin-rich biofilm in the presence of plasma which is highly resistant to attack by the human immune system and to chemotherapy. Varidase, composed mainly of streptokinase, is used for hydrolyzing clots. In this study, we attempted to destroy the biofilm of S. aureus with Varidase and to apply this drug in the treatment of staphylococcal infections. Four clinical isolates were used in the experiments. These organisms formed a several millimeter-thick biofilm on type IV collagen coated coverslips in trypticase soy broth containing 50% human plasma. The biofilm was composed of bacterial cell which adhered to fibrillar fibers and of sediment derived from plasma. 10,000 U/ml of Varidase, the dose which is used clinically, removed the sediment and reduced the number of live bacteria in biofilms to less than 20% of control. 200 U/ml of Varidase was also effective against biofilms of the organisms. An equal combination of Varidase and ofloxacin had an additive effect on the bacteria. The results of this study demonstrate that Varidase is highly effective in destroying biofilms of S. aureus in vitro and suggest that this drug would be useful for treating staphylococcal infections. PMID- 10671762 TI - Efficacy of amikacin, ofloxacin, pefloxacin, ciprofloxacin, enoxacin and fleroxacin in experimental left-sided Pseudomonas aeruginosa endocarditis. AB - The efficacies of amikacin, ofloxacin, pefloxacin, ciprofloxacin, enoxacin and fleroxacin, each as monotherapy, were evaluated in a rabbit model of induced left sided Pseudomonas aeruginosa endocarditis. Therapy started 48 h after infection and lasted 5 days. All agents were given intramuscularly; amikacin at 7 mg/kg/12 h, and each quinolone at 35 mg/kg/12 h. All animals survived except for 1 of the group that received amikacin, and 2 of the untreated control group. No sterile vegetations were found in the untreated group and the group of fleroxacin, while 3 animals from the amikacin, ofloxacin, and enoxacin groups, and 2 from the ciprofloxacin and pefloxacin groups had sterile vegetations. All agents used significantly reduced the number of CFU per gram of vegetation versus untreated controls. Enoxacin and ciprofloxacin were equipotent and more effective than pefloxacin, ofloxacin and amikacin. Fleroxacin had a weaker activity. PMID- 10671763 TI - Bactericidal activity of gatifloxacin (AM-1155) against Pseudomonas aeruginosa and Enterococcus faecalis in an in vitro bladder model simulating human urinary concentrations after oral administration. AB - The bactericidal activity of gatifloxacin, a new 6-fluoro-8-methoxy quinolone, was determined in a dynamic in vitro model mimicking complicated lower urinary tract infection. Strains of Pseudomonas aeruginosa and Enterococcus faecalis with different susceptibility were exposed to changing gatifloxacin concentrations, simulating human urinary concentrations afer oral treatment with 200 mg twice daily for 3 consecutive days. Bacterial numbers of P. aeruginosa (minimal inhibitory concentrations, MIC: < or =32 microg/ml) and of E. faecalis (MIC: 16 microg/ml) were reduced to undetectable levels during exposure. For the strains with lower susceptibility, gatifloxacin showed bactericidal activity, but eradication was not complete. Thus, in a complicated urinary tract infection model, breakpoint MICs of gatifloxacin for uropathogenic organisms were presumed to range from 16 to 32 microg/ml. At least 86% of recent clinical isolates of P. aeruginosa and E. faecalis were inhibited at its breakpoint MIC. These results suggest that gatifloxacin may be useful in the treatment of urinary tract infections. PMID- 10671764 TI - Effects of antibiotics on adherence of Pseudomonas aeruginosa and Pseudomonas fluorescens to A549 pneumocyte cells. AB - The aim of this study was to evaluate the effect of antibiotics at subminimal inhibitory concentrations (sub-MIC) on fluorescent pseudomonas adherence to A549 pneumocyte cells. Pseudomonas fluorescens MF0 isolated from contaminated raw milk and Pseudomonas aeruginosa NK125502 isolated from a cystic fibrosis patient's lung adhered to A549 cells. As previously shown for P. aeruginosa, P. fluorescens bound to A549 cells in a dose-dependent manner over a wide range of bacterial concentrations. Bacterial growth in the presence of polymyxin B or gentamicin at MIC/2 had no effect on the adherence of NK125502 and MF0 to A549 cells. Instead, MIC/2 and MIC/8 of cefsulodin or chloramphenicol decreased the adherence of the two strains. A decrease in MF0 adherence was also observed with cefsulodin at MIC/32. We conclude that, in addition to their antibacterial activity, cefsulodin and chloramphenicol could be effective in preventing Pseudomonas adherence to respiratory epithelium. PMID- 10671765 TI - In vitro effects of meropenem and imipenem/cilastatin on some functions of human natural effector cells. AB - Meropenem, a new carbapenem antibiotic, was assessed to evaluate its effects on some functional parameters of human polymorphonuclear (PMN) and natural killer (NK) cells in comparison with imipenem/cilastatin. Both drugs significantly inhibited PMN phagocytosis and chemotaxis at concentrations of 2,000 and 4,000 microg/ml. They affected PMN microbicidal activity, evaluated against Candida albicans, only at 4,000 microg/ml. A study of the effects of both drugs on peripheral NK populations and the human NK line (NK-92) showed that even at 4,000 microg/ml there was no effect on antitumor activity. These data indicate that meropenem can reduce some PMN antimicrobial functions only at very high concentrations like imipenem/cilastatin, whereas no concentration influenced NK activity. PMID- 10671766 TI - In vitro cytotoxicity of three 4,9-diazapyrenium hydrogensulfate derivatives on different human tumor cell lines. AB - DNA intercalating agents interfere with DNA's role as a template in replication and transcription by inserting an intercalator molecule between adjacent base pairs. We synthesized three potential novel intercalators, 4,9-diazapyrenium hydrogensulfate derivatives: 5, 10-diphenyl-4,9-dimethyl-4,9-diazapyrenium hydrogensulfate (FDAP), 4, 9-dimethyl-4,9-diazapyrenium hydrogensulfate (GDAP) and 2,4,7, 9-tetramethyl-4,9-diazapyrenium hydrogensulfate (MDAP) and tested their biological effects in vitro on four human tumor cell lines (SKBr3: breast carcinoma, HeLa: cervical carcinoma, CaCo2: colon carcinoma and SW620: poorly differentiated cells from lymph node metastasis of colon carcinoma). Cytotoxic effects on cell growth and viability were determined using tetrazolium dye (MTT) assay. DNA synthesis and proliferation of treated cells were studied by the [(3)H]-thymidine incorporation test. DNA fragmentation was analyzed by agarose gel electrophoresis. The growth inhibitory effect was cell-specific and dose dependent. The most pronounced antiproliferative effect was observed on SKBr3 cells for FDAP (10(-5) M) 91.8%, for MDAP (10(-5) M) 85.3% and on SW620 cells for GDAP (10(-5) M) 65.3%. The DNA ladder fragmentation of treated HeLa and SKBr3 cells, as a hallmark of apoptosis, was observed. Based on specific DNA fragmentation, morphological changes (reduced cell volume, round cell shape, condensed chromatin) and growth inhibition of treated human tumor cells we conclude that tested substances induced apoptotic cell death. PMID- 10671767 TI - Adequate levofloxacin treatment schedules for uterine cervicitis caused by Chlamydia trachomatis. AB - The in vivo efficacy of levofloxacin (LVFX), one of the most standard new quinolone antimicrobial agents, in the different treatment schedules of Chlamydia trachomatis uterine cervicitis in women was evaluated. Cervical C. trachomatis was detected by polymerase chain reaction. LVFX at a dosage of 300 mg t.i.d. for 5, 7 and 14 days was orally administered to 18, 33 and 35 Japanese patients, respectively. The eradication rate and the recurrence rate in the different treatment schedules of C. trachomatis were evaluated. The eradication rate in 5-, 7- or 14-day cases was 44.4, 87.9 or 88.6%, respectively. The recurrence rate of 5-, 7- or 14-day cases was 50.0, 0 or 0%, respectively. Seven-day treatment with LVFX is adequate for and effective in C. trachomatis uterine cervicitis. PMID- 10671768 TI - Barrett's esophagus: disregulation of cell cycling and intercellular adhesion in the metaplasia-dysplasia-carcinoma sequence. AB - The incidence of both esophageal adenocarcinoma and Barrett's esophagus, a premalignant condition predisposing to this cancer, is rising rapidly. There is growing evidence that both of these conditions are related to the reflux of acid and bile into the esophagus. This results in inflammation and cell damage which initiates a sequence of events termed the metaplasia-dysplasia-carcinoma sequence in which the squamous epithelium is replaced by columnar epithelium exhibiting increasing degrees of dysplasia and overt malignancy. This sequence of events is underpinned by changes in cell cycling, such as accumulation of p16 and p53 mutations and increased cyclin D1 activity. Progression along this pathway is characterized by changes in intercellular adhesion, in particular, loss of adenomatous polyposis coli, reduced cadherin expression and increased catenin phosphorylation resulting in its nuclear translocation. Herein, we detail these molecular defects and propose how they may interrelate in an ordered progression in the development of esophageal adenocarcinoma. PMID- 10671769 TI - Gastroesophageal reflux disease: prevalence, clinical, endoscopic and histopathological findings in 1,128 consecutive patients referred for endoscopy due to dyspeptic and reflux symptoms. AB - BACKGROUND AND AIMS: Gastroesophageal reflux disease (GERD) reportedly has increased in prevalence while Helicobacter pylori infection and peptic ulcer disease have been on the decrease. The aim of the present study was to examine the prevalence of GERD as well as the clinical, endoscopic and histologic variables that associate with GERD in patients referred for endoscopy. PATIENTS AND METHODS: The study population was drawn from 1,562 consecutive patients referred for endoscopy. The exclusion criteria were previous H. pylori eradication, gastric surgery, anemia and weight loss. Thus 1,128 patients were enrolled in the present study. RESULTS: Of the 1,128 patients, 199 (18%) were referred for endoscopy due to heartburn and/or regurgitation. GERD, defined as chronic (>6 months) heartburn and/or regurgitation with or without erosive esophagitis, Barrett's esophagus, esophageal ulcer or stricture, was detected in 248 (22%) patients. Of the 248 GERD patients, 81 (33%) had endoscopy-negative GERD, but of those aged <50 years (n = 67), 57 (85%) were endoscopy-negative. The overall incidence of GERD was 307 per 100,000 population/year and that of endoscopy-positive GERD 207/100,000/year. The positive and negative predictive values of heartburn and regurgitation for endoscopy-positive GERD were 0.37 (95% CI 0.31-0.44) and 0.90 (95% CI 0.88-0.92), respectively. Independent risk factors for GERD were male sex (OR 1.9, 95% CI 1.3-2.7), previous medication for upper gastrointestinal symptoms (OR 2.7, 95% CI 1.7-4.1), the use of nonsteroidal anti inflammatory drugs (NSAIDs; OR 2.0, 95% CI 1.3-3. 0), histologic esophagitis (OR 2.2, 95% CI 1.5-3.2) and incomplete intestinal metaplasia at the gastroesophageal junction (OR 1.7, 95% CI 1.0-3.1). Chronic gastritis was protective against GERD (OR 0.7, 95% CI 0.5-0.9). No association was observed between GERD and H. pylori infection. The risk of patients aged <50 years (n = 407) of having major lesion (Barrett's esophagus, esophageal stricture, peptic ulcer, esophageal/gastric carcinoma) was significantly lower than that of patients aged >50 years (n = 721; OR 0.5, 95% CI 0.3-0. 9, p = 0.01). CONCLUSIONS: The correlation between reflux symptoms and endoscopy-positive GERD is poor and most GERD patients aged <50 years have endoscopy-negative GERD. The use of NSAIDs is a risk factor for GERD, whereas chronic gastritis, but not H. pylori infection, may protect against GERD. Incomplete intestinal metaplasia at the gastroesophageal junction is associated with GERD. PMID- 10671770 TI - In search of immunogenic Helicobacter pylori proteins by screening of expression library. AB - BACKGROUND: Prevention of Helicobacter pylori infection may help to control related gastritis, peptic ulcer and cancer. Of the possible preventive measures, immunization was successfully employed in various animal studies. However, no immunization protocol has been accepted for humans. A better characterization of the immune response against the pathogen may be required before a human vaccine is developed. AIM: To identify bacterial proteins which induce an immune response in infected humans or H. pylori-immunized rabbits. METHODS: An expression library of H. pylori genes was screened with sera from infected humans and from immunized rabbits. Positive clones were partially sequenced and identified on the basis of a homology search of a H. pylori genome database. Encoded proteins were expressed directly from positive clones and analyzed by SDS-PAGE/Western blot techniques. RESULTS: 114 positive clones were isolated: 79 by screening with human sera and 35 by screening with rabbit sera. Western blot analysis demonstrated that selected clones encoded one or more strongly immunoreactive proteins. 64 clones selected with human sera had no counterparts among clones from screening with rabbit serum. 13 of these clones encoded a total of 21 unknown H. pylori proteins. 17 clones selected with rabbit sera were not immunostained with human sera. They represent 2 various regions of the H. pylori genome which encoded 3 bacterial proteins of unknown function. CONCLUSIONS: Screening of H. pylori expression library identified immunogenic proteins - potential vaccine antigens. PMID- 10671771 TI - 40- to 100-kD protein(s) of Helicobacter pylori stimulate DNA synthesis in epithelial cell lines without affecting apoptosis. AB - BACKGROUND: Previous in vitro studies have demonstrated that water extracts and sonicates of Helicobacter pylori increase DNA synthesis in a small intestinal epithelial cell line. The aim of this study was to identify mitogenic factor(s) in a water extract of a H. pylori strain and to examine their effects on DNA synthesis and apoptosis in vitro. METHODS: IEC-6 and FHs 74 cells were incubated for 24 h with different dilutions of a water extract of H. pylori (cytotoxic strain 88-23) or with 6 protein fractions obtained by gel filtration. Cells were labeled with tritiated thymidine and processed for autoradiography. DNA synthesis was evaluated by the labeling index (LI%). The proportion of IEC-6 cells undergoing apoptosis and/or necrosis was evaluated by flow cytometry using fluorescein isothiocyanate (FITC)-labeled annexin-V and propidium iodide. In vitro caspase activity was also determined as an alternative method for detection of apoptosis. RESULTS: The water extract of H. pylori 88-23 markedly increased DNA synthesis in both epithelial cell lines (p < 0.01). A marked stimulation of DNA synthesis was also observed in IEC-6 cells incubated with fraction II- containing proteins of a molecular weight ranging between 40 and 100 kD (p < 0.01). A lesser stimulation of DNA synthesis was observed in cells incubated with higher concentrations of the other protein fractions (p < 0.01). Neither the water extract of H. pylori 88-23 nor the protein fraction II (40-100 kD) induced apoptosis in IEC-6 cells. CONCLUSION: A water extract of H. pylori 88-23 and a protein fraction containing proteins with molecular weights of 40-100 kD stimulate DNA synthesis in a rat and human small intestinal cell line. Apoptosis was unaffected by the water extract and by protein fraction II, which indicate that the H. pylori-derived mitogen(s) have the capacity to directly enhance epithelial cell proliferation in vitro. PMID- 10671772 TI - Increased mitogen-activated protein kinase activities stimulated with interleukin 1-beta and mechanism(s) of the kinase signaling pathways in rat gastric epithelial cells. AB - Interleukin (IL)-1beta, a multifunctional cytokine, is associated with inflammatory gastric mucosa, but the responses of gastric epithelial cells stimulated by IL-1beta are not known. We determined whether IL-1beta activates the two subfamilies of mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinases (ERKs) and c-Jun NH(2)-terminal kinases (JNKs), in rat gastric epithelial cells (RGM1) using in-gel kinase assays. In addition, we examined the mechanism(s) underlying their signaling pathways and the effect on proliferation of these cells. IL-1beta (0-5 x 10(3) pg/ml) dose dependently induced activation of ERKs (p44ERK and p42ERK) and JNKs (p46JNK and p55JNK) in RGM1 cells; maximal activities were attained with 1,000 pg/ml of IL-1beta. These activities were increased with time, and were maximal 20 min after stimulation with IL-1beta (100 pg/ml). Pretreatment with neutralizing antibody against IL 1beta inhibited IL-1beta-induced activation of ERKs and JNKs. Genistein (100 microM), a tyrosine kinase inhibitor, and GF109203X (2 microM), a protein kinase C inhibitor, inhibited the IL-1beta-induced activation of ERKs and JNKs. Six- hour pre-incubation with IL-1beta inhibited proliferation of these cells by 40% at 24 h of incubation, but the inhibition was recovered at 48 h. These findings suggest that IL-1beta activated ERKs and JNKs in rat gastric epithelial cells and inhibited cell proliferation, possibly via the specific receptor for IL-1beta. Activation of MAP kinases by IL-1beta may be mediated by tyrosine kinase and protein kinase C. PMID- 10671773 TI - Protective effect of lafutidine against indomethacin-induced intestinal ulceration in rats: relation to capsaicin-sensitive sensory neurons. AB - BACKGROUND/AIM: We examined the prophylactic effect of lafutidine, a novel histamine H(2)-receptor antagonist [(+/-)-2-(furfurylsulfinyl)-N-[4-[4 (piperidinomethyl)-2-pyr idyl]oxy- (Z)-2 butenyl]acetamide], on indomethacin induced small intestinal ulcers in rats and investigated the relation of this action to capsaicin-sensitive sensory neurons. METHODS AND RESULTS: Subcutaneously administered indomethacin (10 mg/kg) provoked ulceration in the small intestine, mainly the jejunum and ileum, accompanied by increases in myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) activities as well as the enterobacterial numbers invading the mucosa. Intestinal ulcerogenic response to indomethacin was prevented by 16,16-dimethyl prostaglandin E(2) (10 microg/kg, p.o.) and capsaicin (10 mg/kg, p.o. ) as well as ampicillin (800 mg/kg, p.o.), but not omeprazole (100 mg/kg, p.o.). Likewise, lafutidine (1-10 mg/kg, p.o.), but not cimetidine (100 mg/kg, p.o.), reduced the occurrence of intestinal ulcers in response to indomethacin in a dose-dependent manner, and a significant effect was observed at 3 mg/kg or greater. The protective action of lafutidine as well as capsaicin was almost totally abolished by chemical ablation of capsaicin-sensitive sensory neurons. Both lafutidine and capsaicin significantly suppressed the increases in MPO and iNOS activities as well as enterobacterial numbers in the mucosa. These agents also significantly enhanced mucus secretion in the small intestine. CONCLUSION: These results suggest that lafutidine protects the small intestine against ulceration via stimulation of capsaicin-sensitive sensory neurons. This action may be attributable to inhibition of enterobacterial invasion in the intestinal mucosa, probably by increasing the mucus secretion. PMID- 10671774 TI - Intracellular transport of high-density lipoprotein 3 in intestinal epithelial cells (Caco-2) is tubulin associated. AB - BACKGROUND: A retroendocytotic pathway for high-density lipoprotein 3 (HDL(3)) in cultured intestinal epithelial cell lines has been described. In small intestinal crypt cells and Caco-2, HDL(3) is internalized, transported to lipid droplets and, after solubilization of these lipid droplets, resecreted. In the present study we examined the mechanisms of intracellular transport of HDL(3) in the Caco 2 cell line. METHODS: Apolipoprotein E free HDL(3 )was gold-labeled for transmission electron microscopy and 1, 1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine iodide [DiI(3)] labeled for fluorescence and confocal laser scanning microscopy. For tubulin desintegration Caco-2 cells were incubated with taxol, colchicine and beta- and gamma-lumicolchicine. Tubulin staining was performed using a FITC labeled antibody. Uptake of HDL(3) was quantified by FACS analysis. RESULTS: HDL(3) was rapidly internalized and found to be in contact with lipid droplets in the perinuclear region after 10 min. By transmission electron microscopy a frequent colocalization of HDL(3)-containing vesicles and tubular structures was demonstrated. The close association of HDL(3)-containing vesicles with fluorescence stained tubulin could be confirmed by confocal laser scanning microscopy. Preincubation of the cells with taxol and colchicine did not completely prevent internalization but reduced it during a 2-hour incubation period to less than 50% of the control cells. The transport of DiI(3)-labeled HDL(3) to the lipid droplets in the perinuclear region was almost completely blocked by taxol and colchicine. CONCLUSION: Internalization and intracellular transport of HDL(3) in intestinal epithelial cells (Caco-2) is dependent on a tubulin-mediated mechanism. PMID- 10671775 TI - Regulation of PepT1 peptide transporter expression in the rat small intestine under malnourished conditions. AB - BACKGROUND AND AIMS: Many investigations suggested that peptide nutrition had a clinical advantage for nitrogen absorption. Recently, the cDNA encoding the H(+)/peptide cotransporter PepT1 was cloned. However, the regulatory mechanism of PepT1 expression under malnourished conditions has not been elucidated. The aim of this study was to clarify regulatory mechanisms of PepT1 expression. METHODS: Sprague-Dawley rats were starved for 4 days, semistarved (50% amount of control) for 10 days, or given total parenteral nutrition (TPN) for 10 days. Rats with free feeding were used as control. Among those groups, the changes of PepT1 mRNA level in the jejunal mucosa and PepT1 protein density at the brush-border membranes were examined by Northern blot and by Western blot analysis, respectively. RESULTS: Both starvation and TPN treatment caused a significant decrease in mucosal weight by 41 and 50% respectively. PepT1 mRNA level increased to 179% in the starved group and also to 161 and 164% in the TPN and semistarved groups, respectively. In contrast, sodium-dependent glucose transporter 1 mRNA expression showed no significant change. PepT1 protein density showed similar changes with the mRNA. CONCLUSIONS: PepT1 gene expression was significantly enhanced under the malnourished conditions in spite of atrophic changes of intestinal mucosa. PMID- 10671776 TI - Tumor necrosis factor-alpha in ileal mast cells in patients with Crohn's disease. AB - BACKGROUND: Reports that both intestinal and extraintestinal Crohn's disease (CD) had healed successfully after treatment with anti-tumor necrosis factor-alpha (TNF-alpha) antibody have strengthened the hypothesis that it has a role in the treatment of CD. The macrophage is one source of TNF-alpha. Intestinal mast cells are also thought to have a role in CD, but it is not known if human ileal mast cells express TNF-alpha. AIM: To find out whether TNF-alpha is expressed by mast cells in the ileal wall in CD patients and controls. METHODS: TNF-alpha was sought immunohistochemically in full thickness specimens of ileal wall from patients with CD (histologically normal, n = 9; inflamed, n = 6) and controls (patients with colonic cancer, n = 8). Mast cells were identified by metachromasia and anti-mast cell tryptase immunoreactivity. RESULTS: In all layers of the ileal wall, and in every specimen investigated, mast cells were the main cell type that expressed TNF-alpha immunoreactivity out of the TNF-alpha labelled cells. The number of TNF-alpha- labelled mast cells was greater in the muscularis propria in patients compared with controls, both in uninflamed (1.7 fold, p < 0.05) and in inflamed bowel (4.6-fold, p < 0.002); greater in the submucosa in inflamed compared with uninflamed CD (1.6-fold, p < 0. 01), and less in the lamina propria in inflamed compared with uninflamed CD (0.4-fold, p < 0.05). CONCLUSION: Mast cells are an important source of TNF-alpha in all layers of the ileal wall, and the increased density of TNF-alpha-positive mast cells in the submucosa and muscularis propria may contribute to the tissue changes and symptoms in CD. PMID- 10671777 TI - Current status of minimally invasive treatment options for localized prostate carcinoma. AB - INTRODUCTION: Prostate cancer is the leading malignancy in men today and an increase in detected localized prostate cancers is expected in the years to come. Even though radical prostatectomy is an effective treatment, it is associated with a considerable morbidity in some cases and efforts are made to provide minimally invasive alternative treatment options with equal efficacy but fewer side effects. METHODS: Cryosurgical ablation of the prostate (CSAP), brachytherapy, high-intensity focused ultrasound (HIFU) and radiofrequency interstitial tumor ablation (RITA) were evaluated after a literature review from a Medline Search (1966-1998). Furthermore, personal experience and latest data from the authors were taken into account. RESULTS: All alternative treatments nowadays make use of sophisticated technology, including the latest ultrasound devices for exact planning and monitoring of treatment, leading to increased safety compared to treatments in the 1960s and 1970s. Five-year results of CSAP show a PSA <1 ng/ml in 60% of cases whereas brachytherapy is able to achieve PSA <1 ng/ml in 80% of cases in a selected group. Recent outcome data come close to results of radical prostatectomy series. HIFU and RITA are promising new technologies that proved to be able to induce extensive necrosis, but the follow up is too short to determine their definite places in the treatment of prostate cancer. CONCLUSION: Two alternative treatment options for localized prostate carcinoma, CSAP and brachytherapy, have been studied with a sufficient number of patients and an adequate follow-up. The overall results of brachytherapy are favorable when compared to CSAP and are in the same range as the outcome after radical prostatectomy. HIFU and RITA are relatively new techniques based on sophisticated technology that are very promising at present, but a longer follow up is mandatory. PMID- 10671778 TI - Intrathecal catheter with subcutaneous port for clonidine test bolus injection. A new route and type of treatment for detrusor hyperreflexia in spinal cord-injured patients. AB - INTRODUCTION AND OBJECTIVES: The objective of this study was to assess the feasibitity, technical data and use of intrathecal catheter implantation with subcutaneous port for clonidine test injections and individual evaluation. METHODS: According to approval of the local ethics committee, 9 consecutive SCI patients (6 men, 3 women) had catheter and port implantation between January 1998 and May 1999. All did not respond to systemic drug therapy in combination to self clean intermittent catheterisation (SCIC). Implantation was done under general anesthesia. Needle and catheter were Medtronic Infusion Synchromed Intraspinal catheter (Induratrade mark, 8703W). Clonidine test injections were allowed at D5. RESULTS: There were no complications during operation. Follow-up was 8.2 months (0.5-17). After clonidine bolus injection test and validation, 6 patients decided to have permanent pump implantation, 2 chose other therapies and one did not tolerate clonidine intrathecal injections for blood arterial pressure side effects. CONCLUSIONS: Intrathecal clonidine may represent a useful conservative treatment of both severe bladder hyperreflexia and spinal spasticity. Its short term effects can be individually evaluated through bolus injection in subcutaneous port before definitive pump implantation. PMID- 10671779 TI - Natural history of residual renal stone fragments after ESWL. AB - OBJECTIVE: After extracorporeal shock wave lithotripsy (ESWL), residual fragments (RF) 4 mm or less are usually considered as clinically insignificant. We retrospectively reviewed the natural history and clinical significance of 97 noninfected and isolated RF (4 mm or less) observed 3 months after the last ESWL session on renal tomography. PATIENTS AND METHODS: They represented 83 among 1, 216 patients treated by ESWL over a 9-year period (1989-1997). These RF were mostly localized in the inferior calyx (62%). Median follow-up was 40.6 months (range: 7-96 months). Renal tomography was always performed at the end of follow up. RESULTS: Stone-free status, or a decreased, stable or increased amount of residual stone occurred in 27 (33%), 1 (1%), 24 (29%) and 31 (37%) of the 83 patients, respectively. During this study, 18 patients (22%) were proposed for a complementary treatment related to a size increase of the residual fragments (13 ESWL, 1 retrograde endoscopy, 3 percutaneous nephrolithotomy, and 1 polar inferior nephrectomy). CONCLUSION: The term clinically insignificant should not be employed to describe RF after ESWL. Efforts should be performed to obtain true stone-free status after ESWL. PMID- 10671780 TI - Increase in the prevalence of symptomatic upper urinary tract stones during the last ten years. AB - PURPOSE: In industrialized countries the prevalence of upper urinary tract stones has continually increased during the 20th century, but there are considerable differences between countries and also within the same country. To study whether there is still an increase in the frequency of renal stones, an investigation was undertaken to determine the prevalence of stone formers in a village near Milan, Italy, during two time periods, with an interval of 12 years. MATERIALS AND METHODS: Questionnaires were administered in 1986 and 1998 to all adult (age >25 years) occupants of two random samples of households in the village. Participants were asked whether they had experienced a kidney stone during their lifetime. RESULTS: The overall prevalence of stone formers among males was 6.8% in 1986 and 10.1% in 1998; that among females was 4.9% in 1986 and 5.8% in 1998. In all age classes, the respondents in the 1998 survey more frequently reported a history of stones than in 1986, but the prevalence of renal stones was significantly higher in 1998 than in 1986 only among males aged 31-40 and 51-60 years. The yearly incidence was estimated at 0.4%, with 0.6 and 0.18% in men and women, respectively. CONCLUSIONS: This marked increase in renal stones could be the result of environmental factors such as dietary habits and lifestyle, in particular the influence of an increased consumption of animal protein should be considered. PMID- 10671781 TI - Extracorporeal shockwave lithotripsy for ureterolithiasis in patients with urinary bilharziasis: efficacy and variables that influence treatment outcome. AB - OBJECTIVES: Schistosomiasis affecting the ureter is commonly accompanied by ureteric dilatation with or without ureteric stricture and altered ureteric wall motility that can influence extracorporeal shockwave lithotripsy (ESWL) results. This study attempts to identify variables that may influence the outcome of ESWL in the treatment of ureterolithiasis in patients with urinary bilharziasis. PATIENTS AND METHODS: Forty-three patients with urinary schistosomiasis and ureterolithiasis treated with ESWL were reviewed. The study data include characteristics of patients, stones, urinary tract treated and details of ESWL treatment. RESULTS: Thirty-five patients (81.3%) were stone-free at 3 months. Multivariate analysis with logistic regression identified two significant variables that influenced treatment outcome, namely the presence of ureteric stricture (p = 0.004) and the ESWL voltage (p = 0.003). Ten ureteric strictures were encountered in 9 patients (21%), the majority of these were diagnosed post ESWL when patients failed to pass well-fragmented stones in spite of pre-ESWL evaluation. CONCLUSIONS: In situ ESWL is a safe and effective first line of treatment for urinary stones in bilharzial ureters. The presence of concomitant bilharzial stricture is a significant variable which affects the treatment outcome. Every effort should be made to rule out and deal with possible complicating factors such as ureteric strictures in the pretreatment period. PMID- 10671782 TI - Prevalence and risk factors for urinary incontinence in Italy. AB - OBJECTIVES: To analyze the frequency and risk factors for urinary incontinence (UI) in Italy. METHODS: Eligible for this cross-sectional study were men aged >/=50 years and women aged >/=40, randomly identified among registered subjects of a network of general practitioners during the period March-October 1997. All subjects were invited by telephonic interview to determine the presence of UI, reported by the subjects as loss of urine in the last year. The subjects with UI were further questioned at home for evaluation of the type, degree and frequency of UI episodes. RESULTS: Of the 5,488 subjects interviewed (2,767 women and 2,721 men), 92 (3%) men and 316 (11%) women reported at least one episode of UI during the year before the interview. The frequence of UI increased with age both in men and women, being 2 and 11% in men and women, respectively, aged 50-60 years and 7 and 16% in those aged >/=70. Of the subjects with UI identified, 229 women and 64 men and a group of 289 subjects without UI were questionned at home using a detailed questionnaire. Six and 55% of men and women, respectively, reported stress incontinence, 20 and 12% urge incontinence and 20 and 24% mixed incontinence. The risk of UI increased with body mass index in women. A history of recurrent urinary infection was associated with UI in men and less markedly in women. No association emerged between education, smoking and alcohol or coffee consumption and risk of UI. Parity was directly associated with the risk of UI in women. CONCLUSIONS: The study offers a quantitative estimate of the prevalence of UI and its main risk factors in this Italian population. PMID- 10671783 TI - A comparison of health care financing policies for incontinence products in European countries. AB - OBJECTIVES: This paper offers an overview of the different health care financing policies for incontinence products in 16 European countries and provides health care decision-makers with a framework for positioning their financing policy for incontinence products versus other European countries. METHODS: A questionnaire was sent to institutions or persons acquainted with the health care financing system towards incontinence products in 19 countries. Further details were collected by additional telephone interviews and information from several informants. Three countries did not provide information. RESULTS: Financing systems for incontinence products differ widely from country to country. In all countries, hospitalized incontinent patients are better covered than patients residing in institutions for geriatric care. It is furthermore a common phenomenon that patients living at home receive even less coverage. Moreover, most countries apply a fairly uniform type of financing system, meaning that, once assessed in need, financial coverage is very similar for all patients (i.e. not very much differentiated with respect to the nature and severity of their incontinence problems). CONCLUSION: Given the serious potential impact of incontinence on citizens' quality of life and given the substantial variations in degree of incontinence, most countries could improve their utilization of (scarce) health care resources devoted to incontinence by developing more 'selective' payment policies, whereby reimbursement is 'tailored' to patients' needs. PMID- 10671784 TI - Frequency and determinants of erectile dysfunction in Italy. AB - OBJECTIVE: To analyze the prevalence and risk factors for erectile dysfunction (ED) in Italy in a cross-sectional study. METHODS: Eligible for the study were men aged 18 years or more, randomly identified by 143 general practitioners among their registered patients during the period January 1996 to February 1997. ED was defined as the impossibility to achieve and maintain an erection sufficient for satisfactory sexual performance. RESULTS: Of the 2, 010 men interviewed, 257 (12.8%) reported ED. The prevalence increased with age, from 2% in men aged 18-39 to 48% in those >70 years (tested for trend, p = 0.0001). A history of cardiopathy, diabetes, hypertension, neuropathy, thrombotic/hemorrhagic stroke, peripheral vascular disorders, pelvic/medullary injury and pelvic surgery/radiation all increased the risk of ED. The association of hypertension and diabetes tends to increase the risk of ED. In comparison with nondiabetic and nonhypertensive men, the odds ratio (OR) was 1.4 (95% confidence interval (CI), 0.7-3.2) for hypertensive men without diabetes, 4.6 (95% CI, 1.6-13.7) for diabetic men without hypertension and 8.1 (95% CI, 1.2-55.0) for men with diabetes and hypertension. In comparison with never smokers, the OR of ED was 1.7 (95% CI, 1.2-2.4) for current smokers and 1.6 (95% CI, 1.1-2.3) for ex-smokers and increased with duration of the habit. CONCLUSIONS: The study offers a quantitative estimate of the prevalence of ED and of its main risk factors in Italian men. PMID- 10671785 TI - AMS three-piece inflatable implants for erectile dysfunction: a long-term multi institutional study in 200 consecutive patients. AB - OBJECTIVES: The aim of this study was to assess the longterm mechanical reliability of AMS (American Medical Systems) three-piece inflatable implants and their impact on patient-partner satisfaction in 200 consecutive patients with erectile dysfunction who underwent surgery in five different institutions. METHODS: Patient charts included in the study were collected and extensively assessed to record pre- and intraoperative data and postoperative complications. All patients and 120 partners were then seen often in the office at a mean follow up of 59 months (range 6-130) and they were extensively questioned about function of the device and its impact on the couple's sexual life. RESULTS: At the long term follow-up, 185 patients (92.5%) were still engaging in sexual intercourse with a mean frequency of 1.7/week. Patients and partners reported prosthetic erections as excellent, satisfactory or poor in 96 (48%), 100 (50%) and 4 (2%) cases, and in 20 (17%), 80 (66%) and 20 (17%) cases, respectively. Postoperative sexual activity was considered excellent, satisfactory or poor by 140 (70%), 44 (22%) and 16 (8%) patients and by 34 (28%), 81 (68%) and 5 (4%) partners, respectively. Reasons for patients' complaints included postoperative penile shortening in 60 (30%) cases and poor glandular engorgement in 40 (20%) cases. Partners' main complaint was unnaturalness of the prosthetic erection, a factor reported by 30 (25%) subjects. Complications requiring surgical exploration included infection in 12 patients (6%) and mechanical failure in 8 patients (4%). Kaplan-Meier estimates demonstrated significantly decreased mechanical survival for the Ultrex type of cylinders compared to the CX type of cylinders. CONCLUSIONS: AMS three-piece inflatable implants provide an overall patient and partner satisfaction rate of 92 and 96%, respectively. However, postoperative penile shortening and poor glandular engorgement were the causes of some complaints among the patient population as well as the unnaturalness of prosthetic erection among female partners. In the long-term, mechanically speaking, CX cylinders seem to be more reliable than the Ultrex ones. PMID- 10671786 TI - Glide wires for delayed catheterization of severely obstructed ureters. AB - OBJECTIVE: Failure to retrogradely catheterize an obstructed ureter may require a nephrostomy. We describe an endoscopic technique to catheterize a severely obstructed ureter when the obstruction does not permit passage of a ureteral catheter over a wire. METHODS: Using a cystoscope, a wire is passed beyond the obstruction. When the attempt to insert a ureteral catheter over the wire fails, the wire is left in place, fixed externally to a Foley catheter. A second attempt to insert a ureteral catheter is carried out 24-48 h later. RESULTS: The procedure was performed in 5 patients. Ureteral catheterization, which was initially impossible, was performed successfully and without complications. CONCLUSIONS: Maintaining a wire in place over a short period of time facilitates a subsequent ureteral catheterization. PMID- 10671787 TI - Nephrectomy - indications, complications and postoperative mortality in 646 consecutive patients. AB - OBJECTIVE: To gain information about the indications for and complications of conventional nephrectomy, also to create standards for future evaluation of nephrectomies performed by minimal invasive techniques. METHODS: We present a historical 20 years' series of 646 consecutive nephrectomies performed in the period of 1978-1997. Malignant disease led to the operation in 437 cases, of which 98 were urothelial tumors in the renal pelvis or ureter. 209 kidneys were removed due to benign conditions. The incidence of nephrectomy for benign conditions has declined from 75 in the first 5-year period to 32 in the last. RESULTS AND DISCUSSION: Postoperative complications occurred in 100 patients (15.5%). Nephrectomy for malignant disease had a significantly higher rate of complications than operations for benign conditions (p<0.001), especially hemorrhagic complications and pneumonias were more frequent. There were no differences as a result of the operative approach. Reoperation was carried out in 3.0% of the cases. Overall mortality rate (<30 days) was 3.1%. PMID- 10671788 TI - Comparative study of two different TRUS-guided sextant biopsy techniques in detecting prostate cancer in one biopsy session. AB - OBJECTIVE: The aim of this study was to compare a transrectal ultrasound (TRUS) guided sextant biopsy technique, which puts more emphasis on the apical region of the prostate where most prostate carcinomas (PCs) develop, with the standard sextant biopsy technique. METHODS: A total of 280 patients with suspected PC were included in this analysis. Twelve biopsy cores were obtained from all patients. Six biopsy cores were taken within a lateral parasagittal plane from each lobe at the apex, middle and basis, with an angle of approximately 45 degrees (technique A), and 6 further biopsy cores were taken from the left to the right lateral margin always penetrating the prostate in the apex with the same angle (socalled fan-shaped technique, technique B). Technique A predominantly samples in the sagittal and technique B samples more in the transversal plane with emphasis on the apical region where most PCs develop. The sensitivity in detecting PCs for both techniques was calculated and correlated to the serum prostate-specific antigen (PSA) levels. RESULTS: A total of 72 PCs (25.7%) were diagnosed. We subsequently performed subgroup analysis depending on the serum PSA levels: in patients with a PSA of 10 ng/ml (n = 45) technique A has a PC sensitivity of 93.3% (p = 0.083) and technique B 88.8% (p = 0.023) as compared to our reference standard. The number of positive core biopsies using technique A was superior in 14 cases as compared to 12 with technique B (p = 0.154). In 19 patients the number of positive biopsies was identical. CONCLUSION: Our data suggest that in patients with PSA of 0.05). To our knowledge, this is the first study to document marked expression of eNOS in leiomyomatous tissue, compared to parental myometrium. We also conclude that the mechanism(s) of the growth-promoting effect of estrogen on leiomyomata is mediated by more synthesis of NO. PMID- 10671825 TI - Severe unilateral hydrothorax as the only manifestation of the ovarian hyperstimulation syndrome. AB - BACKGROUND: Unilateral hydrothorax is rarely the sole manifestation of the ovarian hyperstimulation syndrome (OHSS) and is suggestive of the severity of the disease. CASE: A 35-year-old woman presented with mild dyspnea 2 weeks after ovarian stimulation with hMG and hCG and IVF-ET. Chest X-ray revealed a large pleural effusion on the right side. Three consecutive thoracocenteses were needed to drain a total of 6,800 cm(3) of fluid. Following drainage, the respiratory symptoms disappeared. An uneventful pregnancy is in progress. CONCLUSIONS: Thoracocentesis is safe and efficient for the treatment of hydrothorax and may be repeated as often as necessary. Clinicians should be aware of the possibility of unilateral hydrothorax as the sole symptom of OHSS. PMID- 10671826 TI - Magnetic resonance imaging diagnosis of metastasis of chronic myelocytic leukemia to the uterus. AB - We report a case of chronic myelocytic leukemia (CML) in which the initial manifestation of blastic crisis was massive genital bleeding. The bleeding was diagnosed as being caused by CML metastasis to the uterus on the basis of the magnetic resonance imaging findings. PMID- 10671827 TI - Cognitive behavior therapy in panic disorder and comorbid major depression. A naturalistic study. AB - BACKGROUND: There is a lack of evidence about the effectiveness of cognitive behavior therapies (CBT) in settings of routine clinical care as well as in the treatment of panic and comorbid disorders. METHODS: We investigated a group oriented CBT approach for 80 patients with panic disorder including 35 patients with current comorbid major depression. Assessments took place 6 months before treatment, at the beginning and end of treatment, and 1 year later. Structured interviews and multiple clinical self-rating scales were used. RESULTS: Panic patients with comorbid major depression showed higher anxiety-specific and nonspecific pathology. The most striking benefits were in reducing avoidance behavior, while improvements concerning catastrophic beliefs were smaller, but still significant. For most self-rating scale results, patients with and without comorbid depression improved to a comparable degree. However, the end-state functioning of patients with panic disorder and current comorbid depression at admission is significantly lower than for patients with panic disorder alone. CONCLUSIONS: The results point to the necessity to develop and improve treatment approaches for patients with comorbidity of panic disorder and current major depression. PMID- 10671828 TI - Impact of management change and an intervention program on health care personnel. AB - BACKGROUND: There has been a major shift in the organization and responsibility for the provision of geriatric care in Sweden. This was believed to be stressful. We therefore decided to launch a controlled intervention program on health care personnel aimed at enhancing their adaptation and ability to cope. The purpose of this study was to assess the impact of management change on psychosocial parameters of health care personnel and the effects, if any, of a structured intervention program. METHODS: Two separate wards were randomly allocated to be either intervention (I) or control (C) ward. The I-ward personnel were subjected to a psychologist's structured 10-session intervention program for 20 weeks. The program consisted of an initial educational part followed by a practical problem solving discussion part. A structured questionnaire on psychosocial issues was answered by the participants before (0 weeks), immediately after (20 weeks), and 10 weeks after the intervention (30 weeks). RESULTS: There were no significant changes in the psychosocial parameters of the C-ward personnel. In the I-ward, however, there was a significant increase in work demand as well as in positive feelings about work, as compared to the C-ward at the 30-week follow-up. There was also significantly better work comfort in the I-ward. CONCLUSIONS: We did not find any anticipated negative psychosocial effects on health care personnel undergoing an organizational change. However, by offering a structured intervention program to one group of health care personnel, we found some positive psychosocial effects. Future research is needed to pinpoint which factor or factors in the intervention program were the most crucial for the effects to occur. PMID- 10671829 TI - Coping with unfair treatment at work--what is the relationship between coping and hypertension in middle-aged men and Women? An epidemiological study of working men and women in Stockholm (the WOLF study). AB - BACKGROUND: An important hypothesis in psychosomatic medicine is that exposure to psychosocial factors that arouse anger may accelerate the onset of hypertension, particularly if the subject is not allowed to show anger or to deal constructively with the factor that evoked it. For working men and women, being treated in an unfair way at work may be crucial. The present study was designed to answer the question whether the pattern of coping - primarily directed towards the aggressor (open) or directed inwards or towards others (covert) - is associated with hypertension among working men and women. STUDY GROUP: Five thousand seven hundred and twenty working men and women aged 15-64 participated in the study. The participation rate was 76%. METHODS: The coping pattern was studied by means of a Swedish version of a self-administered questionnaire that was originally introduced by Harburg et al. RESULTS: Significant results were confined to the age group 45-54. All analyses were adjusted for age and body mass index. Smoking habits and social class had no effect on the relationships. Low scores (lowest quartile) for open coping tended to be associated with an elevated prevalence ratio (PR) of hypertension both among men (PR 1.3, 95% confidence interval, CI, 0.9-1.7) and women (PR 1.4, 95% CI 1.0-2.0). High scores for covert coping (highest quartile) were associated with an elevated PR of hypertension among men (PR 1.6, 95% CI 1.2-2.2) but not in women. If the analysis was confined to cases without medication, the relationship between a high level of covert coping and high blood pressure was still significant for men. For women, however, no significant findings were made after this operation. Accordingly, the relationship between a low level of open coping and hypertension in women was confined to women with medication. Coping patterns were correlated with psychosocial work environment factors, in particular decision latitude. CONCLUSION: In men, covert coping was associated with prevalence of hypertension. In women, there tended to be a relationship between low scores for open coping and hypertension. PMID- 10671830 TI - Social support and psychological distress in primary care attenders. Ferrara SIMG Group. AB - BACKGROUND: Growing evidence has been collected about the high prevalence of psychological distress in primary care. The role of psychosocial variables, namely the role of support from the patients' interpersonal ties, has not been extensively explored. METHODS: The study investigated the relationship between perceived social support, as evaluated by the Multidimensional Scale of Perceived Social Support, and psychological distress, as measured by the Brief Symptom Inventory, in 1,341 primary care attenders. RESULTS: In comparison with highly supported patients, lowly supported subjects showed higher scores on distress dimensions (e.g. depression, anxiety, phobia) and a higher prevalence of psychological morbidity ('caseness' = 37.3 vs. 11%). CONCLUSION: The findings suggest that, in the setting of primary care, the GPs' assessment of their patients' social support system may be useful in identifying those more vulnerable to psychological stress. PMID- 10671831 TI - The relationship between semantic organization and memory in obsessive-compulsive disorder. AB - BACKGROUND: A variety of evidence suggests that frontostriatal dysfunction is involved in obsessive-compulsive disorder (OCD). This evidence includes both neuroimaging findings and results from studies using neuropsychological assessments. Previous studies have documented nonverbal memory deficits in individuals with OCD, whereas verbal learning and memory were less affected. METHODS: The present study examined both verbal and nonverbal memory in a sample of 17 untreated outpatients with OCD. We also evaluated the effects of encoding strategies which are believed to be mediated by frontostriatal system functioning. RESULTS: OCD patients were significantly impaired in both verbal and nonverbal memory performance. This deficit was correlated with impairments in organizational and semantic clustering strategies at the time of encoding. CONCLUSIONS: Deficits in organizational strategies are consistent with frontostriatal dysfunction models in OCD. PMID- 10671832 TI - Dissociative disorder associated with a colloid cyst of the third ventricle: organic or psychogenic amnesia? PMID- 10671833 TI - A case report showing that subclinical hypothyroidism rather than hyperthyroidism stabilizes rapidly cycling bipolar illness. PMID- 10671834 TI - Stress in trainee anaesthetists. PMID- 10671835 TI - The New Deal - a poor deal for service and training? PMID- 10671836 TI - Airway obstruction with cricoid pressure. AB - Cricoid pressure may cause airway obstruction. We investigated whether this is related to the force applied and to the technique of application. We recorded expired tidal volumes and inflation pressures during ventilation via a face-mask and oral airway in 52 female patients who were anaesthetised and about to undergo elective surgery. An inspired tidal volume of 900 ml was delivered using a ventilator. Ventilation was assessed under five different conditions: no cricoid pressure, backwards cricoid pressure applied with a force of 30 N, cricoid pressure applied in an upward and backward direction with a force of 30 N, backwards cricoid pressure with a force of 44 N and through a tracheal tube. An expired tidal volume of < 200 ml was taken to indicate airway obstruction. Airway obstruction did not occur without cricoid pressure, but did occur in one patient (2%) with cricoid pressure at 30 N, in 29 patients (56%) with 30 N applied in an upward and backward direction and in 18 (35%) patients with cricoid pressure at 44 N. Cricoid pressure applied with a force of 44 N can cause airway obstruction but if cricoid pressure is applied with a force of 30 N, airway obstruction occurs less frequently (p = 0.0001) unless the force is applied in an upward and backward direction. PMID- 10671837 TI - A comparison of patient-controlled epidural analgesia following gynaecological surgery with and without a background infusion. AB - We conducted a randomised, controlled study to investigate the effect of adding a background infusion to patient-controlled epidural analgesia for postoperative pain relief. Forty-two patients scheduled for elective lower abdominal gynaecological surgery received patient-controlled epidural analgesia postoperatively using a mixture of 0.2% ropivacaine and 2.0 microg x ml-1 fentanyl. Patients in group B (n = 20) were given a background infusion of 5 ml x h-1, whereas those in group N (n = 21) were not. There was no difference in pain scores or patient satisfaction scores between the two groups. Patients in group B had a higher total drug consumption (156.8 +/- 34.8 ml vs. 89.5 +/- 41.0 ml; p < 0.0001) and incidence of side-effects (71.4% vs. 30.0%; p = 0.007). Motor blockade during the 24-h study period was also greater in group B (median [range] area under the curve 7.5 [0.0-39.0] h vs. 3.0 [0.0-36.0] h; p = 0.035). We conclude that the addition of a background infusion to patient-controlled epidural anaesthesia is not recommended as it confers no additional benefits. PMID- 10671838 TI - Haemoconcentration by gelatin-induced acceleration of erythrocyte sedimentation rate. AB - Erythrocyte sedimentation rates from 40 suspensions of packed red blood cells in modified fluid gelatin, 4% albumin solution, 6% hydroxyethyl starch and normal saline were measured at room temperature using Westergren's method. The erythrocyte sedimentation rate was extremely high in gelatin and this increase was significant after 10-60 min when compared with the other fluids. Erythrocyte sedimentation rates in albumin, hydroxyethyl starch and normal saline were low and there were no differences between these fluids [erythrocyte sedimentation after 60 min, median (interquartile range): gelatin 128 (111.2-130.0) mm, albumin 2 (1.5-2.0) mm, hydroxyethyl starch 1.5 (1.0-1.6) mm, normal saline 2 (1.9-2.5) mm, p < 0.0001]. The addition of twice the volume of modified fluid gelatin to a volume of red blood cells leads to rapid acceleration of the erythrocyte sedimentation rate. This is caused by increased erythrocyte aggregation, and in clinical practice this effect may be useful for the haemoconcentration of diluted blood from cardiopulmonary bypass circuits or cell-saver autotransfusion in paediatric surgery. PMID- 10671839 TI - Risk factors for pressure sores in the critically ill. AB - Pressure sore development in the critically ill is a well-recognised problem and several risk factors have been put forward as being relevant; however, none has been proved valid in this population. This study examines the effects of specific risk factors for the development of pressure sores in the critically ill. Data on 22 specific risk factors were recorded every 8 h. Of 286 patients who were identified as having a minimum set of three predetermined risk factors, 77 developed pressure sores. Using univariate regression analysis, 18 of the 22 specific factors were identified as being significant (p < 0.05) in the development of pressure sores. Multivariate analysis identified five of these 18 specific risk factors as being independently significant (p < 0.05) in pressure sore development. These five factors were norepinephrine infusion, APACHE II score, faecal incontinence, anaemia and length of stay. PMID- 10671840 TI - Evoked potential monitoring in anaesthesia and analgesia. AB - Electrophysiological monitoring of selected neural pathways of the brain, brainstem, spinal cord and peripheral nervous system has become mandatory in some surgery of the nervous system where preventable neural injury can occur. Evoked potentials are relatively simple methods of testing the integrity of various aspects of the nervous system. This review covers the variety of evoked potentials that can be monitored and outlines the principles of their measurement. Their use in specific situations and how factors such as anaesthesia might affect them is presented. PMID- 10671841 TI - Bispectral index monitoring: comparison of two types of electrode. AB - Bis-monitoring is a new method of monitoring anaesthetic depth. Bis-monitoring is easy to perform, but the Bis-monitor and the original, disposable electrodes are expensive. The aim of this study was to determine whether the original Zipprep electrodes could be replaced by the much cheaper electrocardiogram electrodes. We compared bispectral index measurements, conducted using both types of electrode in the same patients before anaesthesia, and during light and deep anaesthesia, in patients randomly allocated to receive either sevoflurane or propofol anaesthesia. We found very good agreement between the measurements from the two different sets of electrodes. The impedance in the electrocardiogram electrodes was higher than in the Zipprep electrodes, but this did not affect the bispectral index. No other problems with either type of electrode were detected. It is concluded that Zipprep electrodes can be replaced by electrocardiogram electrodes in normal clinical practice. PMID- 10671842 TI - Evaluation of a needle-free injection system for local anaesthesia prior to venous cannulation. AB - We evaluated a single-use, disposable, carbon-dioxide-powered, needleless injector (J-Tip, National Medical Products Inc., CA, USA), which is claimed to deliver a virtually painless, subcutaneous injection. Seventy-two patients undergoing various types of surgery had a large-bore intravenous cannula inserted prior to induction of general anaesthesia. Three minutes beforehand, a subcutaneous injection of 0.3 ml of 1% plain lidocaine was administered. Subjects were randomly allocated to receive the lidocaine either by the needleless injector or from a conventional syringe and a 25 G needle. Pain scores were recorded on injection of the lidocaine and on insertion of the cannula. There was significantly less pain on injection with the needleless injector than with the 25 G needle (p < 0.001) but, surprisingly, there was more pain on cannulation (p < 0. 001). We conclude that the device certainly delivers a less painful subcutaneous injection than a 25 G needle, but perhaps provides less effective skin anaesthesia for venous cannulation at sites where the subcutaneous space is small; its use might be better suited to areas where the subcutaneous space is deeper. PMID- 10671843 TI - An assessment of the thermal safety of microwave warming of crystalloid fluids. AB - We performed an in vitro study to determine the thermal safety of a domestic microwave to warm intravenous crystalloid solutions. Five-hundred-millilitre bags of crystalloid, randomly allocated to groups which differed in power setting, timer setting and whether or not agitation was performed after warming, were heated in a microwave oven to a calculated temperature of 39 degrees C. Timer accuracy was checked by stopwatch. Bag temperature was measured using an infrared tympanic temperature probe and fluid temperature was measured with an in-line thermocouple. Mean times measured by stopwatch were higher than set. No in-line temperatures reached 40 degrees C. Wider overall ranges and a higher mean were found with the tympanic probe compared with in-line temperature measurement. There were significant differences between the in-line temperatures of shaken and unshaken bags at each power setting, but not when groups were added together. There was no change in colour or odour of bags or fluid. One bag developed a pinhole leak when the packaging was removed. PMID- 10671844 TI - Target-controlled infusion of propofol and remifentanil combined with bispectral index monitoring for awake craniotomy. AB - We describe the target-controlled administration of propofol and remifentanil, combined with monitoring of the bispectral index, during an awake craniotomy for removal of a left temporo-parietal tumour near the motor speech centre. Target concentrations of the two drugs were adjusted according to the patient's responses to painful stimuli and surgical events, and the need for speech testing. Allowing the effect-site concentrations of propofol and remifentanil to decrease during surgery allowed the performance of cortical speech mapping and the testing of the patient's ability to speak. Although the bispectral index was not used as a guide for the administration of the drugs, its value correlated better with the patient's responsiveness than did the predicted effect-site concentrations of propofol. Side-effects, comprising hypotension, respiratory depression and airway obstruction, were related to rapid increases in drug infusion rates and were easily managed. PMID- 10671845 TI - Towards a pain-free venepuncture. AB - A randomised, prospective trial was conducted to assess the efficacy of various means of alleviating the pain of subcutaneous lidocaine infiltration. One hundred and twenty-two patients were randomly allocated to different groups to receive buffered lidocaine 1%, warmed lidocaine 1% or infiltration by the counter irritation technique. A visual analogue pain score was recorded at different stages of cannulation and results showed that pain scores were significantly lower in the group receiving buffered lidocaine 1% (p < 0.02) and in the counter irritation group (p < 0.05). Thus buffering lidocaine 1% and administration of lidocaine 1% by the counter-irritation technique is effective in relieving the pain of lidocaine infiltration. PMID- 10671846 TI - The effect of cricoid pressure on the cricoid cartilage and vocal cords: an endoscopic study in anaesthetised patients. AB - Cricoid pressure is used to protect the lungs from contamination with gastric contents during tracheal intubation. We studied the effect of cricoid pressure applied with a yoke on 30 anaesthetised patients examined fibreoptically through a laryngeal mask airway. We assessed the effect of 20, 30 and 44 N on the internal appearance of the cricoid and vocal cords. Difficulty in ventilation was also recorded. At 44 N, cricoid deformation occurred in 27/30 patients (90%) and 15/30 (50%) had cricoid occlusion [13/30 (43%) had cricoid occlusion at 30 N and 7/30 (23%) at 20 N]. Associated difficulty in ventilation was present in 15 patients (50%) and 18/30 (60%) had vocal cord closure with associated difficult ventilation, at forces up to 44 N. Cricoid occlusion was unrelated to age and body mass index but females were at greater risk. Orthodox values of cricoid pressure, applied with a yoke, may produce obstruction at the level of the cricoid cartilage or vocal cords, with implications for tracheal intubation and ventilation by mask. PMID- 10671847 TI - Effect of clonidine premedication on haemodynamic responses to fibreoptic bronchoscopy. AB - The usual haemodynamic response to fibreoptic bronchoscopy is an increase in heart rate and blood pressure. We therefore compared, in a prospective, randomised, double-blind study, the effect of two doses of oral clonidine premedication (150 microg or 300 microg) with placebo (control group) on the haemodynamic alterations in 62 patients who underwent elective fibreoptic bronchoscopy. Significant increases in blood pressure and heart rate were observed during fibreoptic bronchoscopy only in the control group. Clonidine 150 microg blunted the haemodynamic response to fibreoptic bronchoscopy (p < 0.05). Significant decreases in systolic blood pressure (< 90 mmHg) were observed in all patients premedicated with 300 microg clonidine. Throughout the study nine patients (75%) in the 300 microg clonidine group were treated at least once for hypotension. Compared with the control group, time to awakening was significantly longer only in patients premedicated with 300 microg clonidine. In conclusion, premedication with 150 microg oral clonidine attenuates haemodynamic responses to fibreoptic bronchoscopy, without causing excessive haemodynamic depression and sedation. These data encourage the administration of clonidine as premedication in patients undergoing fibreoptic bronchoscopy, particularly in those with, or at risk for, coronary artery disease. PMID- 10671848 TI - A new practical classification of laryngeal view. AB - A new practical classification of laryngeal view at laryngoscopy is presented and evaluated. The best laryngeal view obtained with or without anterior laryngeal pressure is recorded. The laryngeal view is easy (E) when the laryngeal inlet is visible. The view is restricted (R) when the posterior glottic structures (posterior commissure or arytenoids) are visible or the epiglottis is visible and can be lifted; this includes some grade 2 and some grade 3 views as classified by Cormack and Lehane. A difficult (D) view is present when the epiglottis cannot be lifted or when no laryngeal structures are visible. Five hundred patients were studied. Laryngoscopy, with the patient anaesthetised and paralysed, was performed with a Macintosh laryngoscope. If the vocal cords were not visible, a gum elastic bougie was used to aid intubation. Other aids were used only if this did not allow intubation. Each laryngeal view was graded according to the new classification and that of Cormack and Lehane. Intubation was timed and the equipment needed to facilitate intubation was recorded. The new classification stratified increasing difficulty with intubation (time for intubation longer and increasingly complex methods needed) better than the Cormack and Lehane classification. The new classification is as sensitive and more specific than the Cormack and Lehane classification in predicting difficult intubation. It is also more sensitive and more specific in predicting easy intubation. PMID- 10671849 TI - Haemodynamic changes during laparoscopic anterior fundoplication measured by trans-oesophageal Doppler ultrasound. AB - We investigated the cardiovascular effects of pneumoperitoneum and steep head-up tilt during laparoscopic fundoplication using an intra-oesophageal Doppler ultrasound probe. Repositioning of the probe proved sufficient to maintain the signal throughout the procedure despite the pneumomediastinum. There was a statistically significant increase in mean arterial blood pressure and a fall in stroke distance but not in systemic vascular resistance. Increasing or decreasing the blood pressure with drugs improved stroke distance. The oesophageal Doppler ultrasound proved a satisfactory method for assessing cardiovascular changes during fundoplication. PMID- 10671865 TI - BJU international volume 85, number 2, january 2000 european urology update series 2000:1 prostatitis: lessons from the 20th century PMID- 10671850 TI - Pretreatment with ketorolac and venous occlusion to reduce pain on injection of propofol. AB - We performed a randomised, double-blind, prospective trial to discover whether intravenous ketorolac 10 mg made up to 2 ml with saline, with or without venous occlusion for 2 min, reduces the pain on injection of propofol. In 90 patients, pain scores were obtained during injection of propofol following pretreatment of the vein with saline, ketorolac or ketorolac with venous occlusion. Pain on injection of ketorolac was more common than with saline (p = 0.02). The incidence of severe pain following propofol was reduced by ketorolac with venous occlusion (p = 0.019) compared with saline or ketorolac without venous occlusion. There was no difference in venous sequelae at 7 days postoperatively between the groups. Our results suggest that pain on injection of propofol may be related to release of local kininogens and that nonsteroidal anti-inflammatory drugs may have a role in reducing that pain. PMID- 10671866 TI - Prostatitis: lessons from the 20th century. PMID- 10671867 TI - Acute urinary retention. PMID- 10671868 TI - Transurethral electrovaporization and vapour-resection of the prostate: an appraisal of possible electrosurgical alternatives to regular loop resection. PMID- 10671869 TI - Human papillomavirus and urological tumours: II. Role in bladder, prostate, renal and testicular cancer. PMID- 10671870 TI - Drugs and intravenous contrast media. PMID- 10671871 TI - Unilateral pedal lymphography in patients with filarial chyluria. AB - OBJECTIVE: To evaluate the usefulness of unilateral pedal lymphography in patients with filarial chyluria. PATIENTS AND METHODS: Of 114 patients with filarial chyluria, all underwent lymphography (unilateral pedal in 106) and 55 underwent selective ureteric sampling for chyle. RESULTS: Unilateral pedal lymphography in the 106 patients detected lymphaticorenal fistulae (LRF) in 104 (98%). Lymphatic crossover was seen in all 106 patients, from the second sacral segment to the first lumbar segment. The most frequent crossover site was at the L5 level (87%). There was complete correlation between the side of LRF and the side of chyluria as assessed by selective ureteric sampling. CONCLUSION: Unilateral pedal lymphography can detect LRF via lymphatic crossover even when it is on the opposite side from that injected with contrast agent. The advantages of unilateral lymphography over bilateral procedures are that it is easy to identify crossover channels, and the discomfort for the patient is reduced because there are fewer incisions and it is quicker. Unilateral lymphography is recommended as the initial method when lymphography is indicated in filarial chyluria. PMID- 10671872 TI - Is bowel preparation useful before radiography of the renal tract in patients with spinal cord injury? AB - OBJECTIVES: To assess, in a blinded study, the usefulness of bowel preparation in improving the quality of radiography of the renal tract in patients with spinal cord injury (SCI). PATIENTS AND METHODS: Plain abdominal radiographs of 56 patients with SCI were selected; 24 of the patients had received bowel preparation and 32 had not. The films were independently assessed by one radiologist and one urologist unaware of the treatment and identity of the patients. Each film was divided into five regions of interest and scores of 1-4 (1 for least and 4 for best visibility) assigned to each area. In films with a low aggregate visibility score (/= 5.5 points on a scale of 0-10, and were included in a further analysis. Based on levels-of-evidence criteria, there is weak evidence to suggest that bladder (re)training is more effective than no treatment (controls), and that bladder (re)training is better than drug therapy. Stimulation types and parameters in the studies of electrical stimulation were heterogeneous. There is insufficient evidence that electrical stimulation is more effective than sham electrical simulation. To date there are too few studies to evaluate effects of PFM exercise with or without biofeedback, and of toilet training for women with UUI. CONCLUSION: Although almost all studies included reported positive results in favour of physical therapies for the treatment of UUI, more research of high methodological quality is required to evaluate the effects of each method in the range of physical therapies. PMID- 10671879 TI - The use of cadaveric fascia lata in the treatment of stress urinary incontinence in women. PMID- 10671880 TI - The immunohistochemical assessment of occult regional lymph node metastases in patients with T3pN0M0 prostate cancer before definitive radiotherapy. AB - OBJECTIVE: To detect occult regional lymph node metastases in patients with T3pN0M0 prostate cancer not recognized by routine haematoxylin and eosin staining, and to evaluate the clinical relevance of this finding. PATIENTS AND METHODS: Formalin-fixed and paraffin-embedded pelvic lymph nodes (1118) from 92 patients were evaluated by immunohistochemistry using antibodies for prostate specific antigen (PSA) and pancytokeratin (AE1/AE3). Of the tumours, 14% were well, 69% moderately and 17% poorly differentiated. The extent of tumour was categorized as T3pN0M0 in all patients, who were referred for definitive radiotherapy after pelvic staging lymphadenectomy. The median (range) serum PSA value before treatment was 18.5 (0.4-342) microg/L. After radiotherapy, the patients were followed by assessing biochemical progression, pelvic recurrence and/or development of distant metastases. The median (range) observation time for all patients was 61 (16-136) months. RESULTS: Occult lymph node metastases were detected in four (4.4%) of the 92 patients. Patients with or without occult metastases had similar serum PSA levels and histological grades. None of the four patients with occult metastases progressed, compared with 37 of the 88 (42%) with no such metastases. CONCLUSION: Using immunohistochemistry the detection rate of occult lymph node metastases in patients with T3pN0M0 prostate cancer is low. The occurrence of such metastases is probably unrelated to the serum PSA value before treatment. The short-term outcome of patients subsequently treated with definitive radiotherapy does not seem to be associated with the finding of occult lymph node metastases, but long-term follow-up is needed. So far, the results do not justify the search for occult lymph node metastases as a routine procedure in these patients PMID- 10671881 TI - Elevated serum vascular endothelial growth factor in patients with hormone escaped prostate cancer. AB - OBJECTIVE: To investigate the role of serum vascular endothelial growth factor (VEGF) in the assessment of patients with prostate cancer. Patients, subjects and methods Serum from 78 men was assayed for VEGF using a commercially available enzyme-linked immunosorbent assay kit. Forty-eight patients had a histopathological diagnosis of prostate cancer (16 local disease, 32 metastatic), nine had benign prostatic hyperplasia (BPH) and 21 were healthy controls. RESULTS: The mean serum VEGF level was significantly higher in patients with hormone-escaped prostate cancer than in all other groups (P /= 0.2 ng/mL above the PSA nadir. RESULTS: The complications were rectal pain (26%), urinary infection (3%), scrotal oedema (12%), haematuria (6%) and incontinence (6%). Complication rates were higher in those patients who failed after radiation therapy than in those who did not receive radiation (incontinence 11% vs 0%, rectal pain 37% vs 12%) but the difference was not statistically significant. Twenty-two patients (63%) had an undetectable serum PSA nadir (< 0.1 ng/mL) after cryotherapy and 30 (84%) patients had a PSA value of < 1.0 ng/mL. After a mean follow-up of 8.3 months (range 0.2-18), nine patients had biochemical recurrence. The biochemical recurrence-free survival (BRFS) was 70% at 9 months. Patients who had an undetectable PSA nadir had a statistically higher BRSF at 9 months than did patients who had a detectable PSA nadir (89% vs 55%, respectively, P = 0.03). Similarly, patients with a preoperative serum PSA level of < 10 ng/mL had a statistically higher BRFS than patients who had a PSA level of > 10 ng/mL (86% vs 42% at 9 months, P < 0.001). CONCLUSION: A PSA level before cryotherapy of < 10 ng/mL and an undetectable PSA nadir after cryotherapy were associated with the highest BRFS. Cryoablation of the prostate, with low morbidity, seems to be a viable option in managing patients by salvage therapy after radiation therapy and for the primary treatment of clinically localized prostate cancer. PMID- 10671883 TI - Radical retropubic prostatectomy: time trends, morbidity and mortality in Japan. AB - OBJECTIVES: To assess the time trends, morbidity and mortality of contemporary anatomical radical retropubic prostatectomy (RRP) in a multi-institutional study in Japan, where RRP has become more popular in the last decade. PATIENTS AND METHODS: Between January 1991 and August 1998, 638 patients underwent RRP at seven urological centres in Japan. Major complications (within 30 days of surgery) and the 30-day mortality were reviewed retrospectively. Of the patients, 12.9% were < 60 years old, 56.3% were 60-69 years old and 30.9% were >/= 70 years old (median age 67). Results The number of RRPs increased markedly, by more than sevenfold, from 1991-92 to 1996-97, mainly because there were more patients undergoing RRP in their sixth decade. The contribution of T1c disease increased in absolute and relative terms, from 13.9% in 1991-92 to 37.9% in 1997-98. Over time, the mean blood loss and the allogeneic transfusion rate decreased steadily. There was a trend toward more favourable outcomes for pathological variables (an increased percentage of organ-confined disease, decreased margin positivity and a decreased incidence of positive lymph node metastasis). The most common complications were wound-related (7.5%), or anastomotic leakage (4.1%). Major cardiopulmonary complications occurred in only two patients (0.31%, both pulmonary embolisms). One patient died from cerebral haemorrhage within 30 days of surgery, giving a mortality rate of 0.16%. CONCLUSION: s This study indicates a trend towards selecting patients most likely to benefit from RRP. Although the procedure is technically demanding, it can have an acceptably low rate of early complications, little mortality and need for allogeneic transfusion. The assessment of morbidity suggests a lower incidence of catastrophic thrombo embolic and cardiac complications in Japanese patients than in Western men. The present data may be useful in decision-analysis models evaluating the role of therapy for Asian men with early-stage prostate cancer. PMID- 10671884 TI - Genital self-mutilation: there is no method in this madness. PMID- 10671885 TI - The histological extent of the local spread of carcinoma of the penis and its therapeutic implications. AB - OBJECTIVE: To explore the possibility of reducing the margin of clearance at surgery for carcinoma of the penis without causing an increase in the incidence of local tumour recurrence, so that the functional and cosmetic compromise associated with penectomy might be minimized. PATIENTS AND METHODS: Sixty-four patients underwent partial or total penectomy based on the extent of tumour. The specimens were evaluated histologically for grade and for proximal microscopic extensions beyond the grossly visible tumour margin, by examining serial proximal 5 mm sections. The histological grade of the lesion was correlated with its clinical site, morphology and proximal microscopic spread. Differences were assessed using the chi-squared test. RESULTS: Of 64 tumours, 31% were grade 1, 50% grade 2 and the remaining 19% grade 3. Higher grade lesions were more likely to involve the penile shaft. The maximum proximal histological extent was 5 mm for grades 1 and 2, and 10 mm for grade 3 tumours; there was no discontinuous spread. CONCLUSIONS: Histological grading is mandatory in the management of carcinoma of the penis. A 10-mm clearance is adequate for grade 1 and 2 lesions, and 15 mm for grade 3 tumours. This approach would qualify more patients for partial rather than total amputation; the residual length of the penis would then be cosmetically and functionally more acceptable. PMID- 10671886 TI - Testicular adenomatoid tumours: clinical and ultrasonographic characteristics. AB - OBJECTIVE: To determine if benign testicular tumours can be identified clinically and ultrasonographically before surgery. PATIENTS AND METHODS: The clinical and ultrasonographic findings of six patients with testicular adenomatoid tumours and 16 with testicular germ cell tumours were assessed retrospectively and compared. RESULTS: All adenomatoid tumours were characterized on physical examination by a well-defined, painful and unfixed nodule, contrasting with the painless and ill defined malignant lesions. On ultrasonography, of the six adenomatoid tumours, two appeared hypoechoic, one hyperechoic, two isoechoic and one was normal, whereas none of the 16 tumours appeared normal or isoechoic. CONCLUSIONS: While isoechogenicity was not apparent in the group of testicular malignancies, the two groups had hypo- and hyperechoic patterns. Small, superficial, painful and unfixed testicular tumours that appear isoechoic on ultrasonography should be biopsied through an inguinal approach, with frozen sections assessed, instead of the patients undergoing immediate radical orchidectomy. PMID- 10671887 TI - Low-grade left varicocele in patients over 30 years old:the effect of spermatic vein ligation on fertility. AB - OBJECTIVES: To evaluate the effect of spermatic vein ligation in patients over 30 years old and with low-grade left varicocele, and thus help to establish whether such patients might benefit from surgery. PATIENTS AND METHODS: A randomized study was conducted on 68 infertile patients (30-38 years old) with evidence of sperm abnormalities and who had low-grade varicocele (grade I according to Hirsch), comparing left spermatic vein ligation with no treatment. The outcome was assessed by standard sperm analysis and eventual paternity. RESULTS: There was no improvement in sperm quality in either of the groups one year after surgery, and no significant difference in paternity. CONCLUSIONS: Left spermatic vein ligation for low-grade varicocele in patients more than 30 years old cannot be recommended. PMID- 10671888 TI - The role of the renal resistive index ratio in diagnosing obstruction and in the follow-up of children with unilateral hydronephrosis. AB - OBJECTIVE: To assess the role of the renal resistive index ratio (RIR) in discriminating equivocal upper urinary tract dilatation in children, and thus in establishing the need for surgery, in comparison with traditional diagnostic tools. PATIENTS AND METHODS: The study comprised 40 children with unilateral hydronephrosis unrelated to vesico-ureteric reflux, posterior urethral valves, megaureter or a duplex system. In all patients one or more of the following 'indices of obstruction' were positive; an anteroposterior renal pelvic diameter of >20 mm, a half-time diuretic 'washout' (T/2) of > 20 min, a separate renal function of < 40%, and symptoms of obstruction (pain, sepsis). All these variables were measured on admission and after a mean (range) follow-up of 9 (2 24) months. After this period, all patients who were symptomatic or with two or more of the variables above the normal range were considered as obstructed and underwent a dismembered pyeloplasty. The variables were then re-assessed 6 months after surgery. The RIR was evaluated using duplex Doppler ultrasonography with a 3.5-5 MHz transducer, by the same operator. Differences between obstructive and unobstructive unilateral hydronephrosis were estimated from the mean values of the variables assessed and Student's t-test used to determine significant differences. The correlation between the T/2 and RIR before and after surgery was also evaluated. RESULTS: During follow-up the RIR was abnormal in 27 of 30 patients with hydronephrosis who were considered to be 'obstructed'. Twenty-three of these patients, selected for surgery, had a positive diuretic renogram; 11 had loss of differential renal function and 16 had recurrent clinical symptoms. There were significant differences in the mean RIR and T/2 between obstructed and unobstructed patients. Six months after dismembered pyeloplasty, the RIR returned to normal in all patients except three in whom it was previously > 1.1. The diuretic renogram, if initially showing pathology, always became normal. The RIR did not change in patients with an unchanged and severe loss of differential renal function before and after surgery. CONCLUSIONS: In this study the RIR was a good index of obstruction in children with unilateral hydronephrosis and it correlated well with the results of diuretic renography. The specificity of the RIR was reduced whenever there was severe renal damage. PMID- 10671889 TI - The efficacy and parenchymal consequences of extracorporeal shock wave lithotripsy in infants. AB - OBJECTIVES: To determine the efficacy of extracorporeal shock-wave lithotripsy (ESWL) in young children and to evaluate, using renal scintigraphy, any possible adverse effects on renal parenchyma. PATIENTS AND METHODS: From January 1991 to October 1998, 19 infants (aged 5-24 months) underwent ESWL for kidney urolithiasis using a Sonolith 3000 (14 kV, Technomed Corp, Lyon, France) or a Nova (14-20 kV, Direx Medical Systems, Paris, France) lithotripter. The treatment and its effects were evaluated using a physical examination, conventional imaging (plain abdominal X-ray and ultrasonography) and renal scintigraphy 24 h before ESWL and again at least 6 months after the last session of treatment. RESULTS: Ten children were rendered stone-free by ESWL after one session and 18 after two sessions. At the follow-up (8 months to 8 years, mean 36 months) no hypertension was recorded and no acquired parenchymal damage was detected with conventional imaging. No scars or significant variation of differential function attributable to ESWL were identified on renal scintigraphy. CONCLUSION: ESWL is clearly effective for treating infant urolithiasis. There were no renal parenchymal lesions associated with ESWL, even in previously damaged kidneys or after the treatment of staghorn calculi. A long-term follow-up (assessing blood pressure) is mandatory and renal scintigraphy before and 6 months after ESWL in infants is recommended to confirm these results in a larger series. PMID- 10671890 TI - Schistosoma haematobium infection in children in Britain. AB - OBJECTIVE: To highlight the existence of Schistosoma haematobium in certain ethnic minority groups in Britain and in English citizens who have recently visited Africa and the Middle East, so that general practitioners and paediatric nephrologists/urologists are aware of its occurrence and consider it among the differential diagnoses in children presenting with haematuria. PATIENTS AND METHODS: Over a 2-year period, six consecutive boys (mean age 13.5 years, range 8 15) presented with haematuria and were subsequently diagnosed to be infected with S. haematobium. All patients were from Africa and had recently visited their native country. There had all reported paddling in freshwater lakes and streams. RESULTS: Dysuria and haematuria was noted 2-3 months after the infection. Terminal urine samples taken after exercise at midday were positive for S. haematobium ova. Praziquantel anti-schistosomal chemotherapy was effective in treating the infection. CONCLUSION: S. haematobium infection is treatable in the early stages and the changes are reversible before the development of fibrotic lesions, which may result in anatomical obstruction. A terminal urine sample taken at midday after exercise was diagnostic in showing Schistosoma ova in all cases. This infection must be considered in the differential diagnoses of haematuria in some ethnic minority British citizens and in those Britons who have visited Africa or the Middle East in the recent past. PMID- 10671891 TI - In vitro contractile responses of detrusor to carbachol and neurokinin A, in children with recurrent urinary tract infection or day wetting. AB - OBJECTIVE: To investigate whether a history of recurrent urinary tract infection (UTI) and/or the presence of day-wetting/urge symptoms might influence the contractile responses to the cholinergic agonist carbachol or to the sensory neuropeptide neurokinin A (NKA) in the urinary bladder in children. PATIENTS AND METHODS: Small detrusor strips were taken from the margin of the cystotomy incision of the bladder dome during surgery to correct vesico-ureteric reflux (VUR) in 62 children (aged 4 months to 12 years) or for unrelated bladder conditions in five controls (aged 3 months to 13 years). Concentration-response curves to carbachol and NKA were constructed using organ-bath techniques, and results compared for age, sex, weight of the detrusor strip, UTI history or day wetting syndrome. RESULTS: The contractile responses to NKA were no different for any of the features investigated. The contractile response to carbachol and NKA in detrusor from control and VUR patients was not significantly different. The children with a history of UTI were significantly older than those without. The contractility in response to carbachol was greater in older girls (aged 4-12 years) than younger girls (< 4 years) and than in all boys (< 4 years and 4-12 years; ANOVA P = 0.013). The mean (SEM) maximum contractile response to carbachol in the group of 20 young children (4-30 months) with previous UTI was 3.0 (0.3) g, whereas the maximum response in the age-matched group of 11 without UTI was 1.8 (0.3) g (P = 0.046). There were no significant differences in maximum responses between those with day-wetting and those without (aged > 4 years), although there was a significant difference in pD2 value, at 6.19 (0.13) and 5.58 (0.14), respectively (P = 0.018). CONCLUSION: Carbachol produced a larger contractile response in detrusor from children with a history of UTI than from those without, indicating possible alterations in muscarinic receptor characteristics. An increased sensitivity to muscarinic stimulation in day wetting children was also suggested, whereas NKA is unlikely to be involved in any of these pathophysiological conditions. PMID- 10671892 TI - Apoptosis in the erectile tissues of diabetic and healthy rats. AB - OBJECTIVE: To compare the frequency of apoptosis in the erectile tissue of chronic diabetic and healthy rats. MATERIALS AND METHODS: Fourteen chronic diabetic and 10 healthy Sprague-Dawley rats were killed, their penises harvested and stored at -70 degrees C until staining and flow cytometric analysis for apoptosis. A cell suspension was obtained from the penile tissue by scraping the inside of the cavernosum with a scalpel and filtering through a mesh. Samples of the cell suspension (0.5 x 106 cells) were stained with Annexin V (an indicator of apoptosis) and propidium iodide (PI, which stains dead cells), incubated for 15 min at room temperature and analysed by flow cytometry. The DNA content was also analysed in each sample. RESULTS: In normal erectile tissue, a mean of 6.2% of cells were stained with Annexin V, while only 2.7% were stained with PI; DNA content analyses showed 7.5% were hypodiploid cells. In diabetic rats 19.5% of cells were stained with Annexin V and 5.2% with PI; 22.9% of cells were hypodiploid. CONCLUSION: The ratio of apoptotic cells in the erectile tissues of diabetic rats was significantly greater than in normal rats. The high rate of apoptosis in diabetic rats may play a role in the pathophysiology of erectile dysfunction. PMID- 10671893 TI - Do experimentally induced ipsilateral testicular torsion, vas deferens obstruction, intra-abdominal testis or venous obstruction damage the contralateral testis through a common mechanism? AB - OBJECTIVE: To evaluate if various conditions affecting the ipsilateral testis which also damage the contralateral testis share a common pathway for their effects. MATERIALS AND METHODS: The study comprised five groups of 10 adult rats which underwent surgery to produce (on their left sides); group 1, a sham operation (control); group 2, testicular torsion; group 3, vas deferens obstruction; group 4, an intra-abdominal testis; and group 5, venous obstruction. The ipsilateral and contralateral testes were harvested 4 weeks after surgery. The relative proportions of haploid cells, the mean seminiferous tubular diameter (MSTD), mean testicular biopsy scores (MTBS), and lactate and hypoxanthine levels were determined and compared. RESULTS: The proportions of haploid cells in the ipsilateral and the contralateral testes of groups 2-5 were significantly lower than those of the corresponding testes of the control group. The MSTD and MTBS of the ipsilateral testes in groups 2-5 were also significantly lower than the ipsilateral testes of controls and the contralateral testes within the same groups. While the MSTD and MTBS of the contralateral testes of groups 1 and 5 were not significantly different, those of the contralateral testes of groups 2-4 were significantly less than that of group 1. The lactic acid and hypoxanthine levels of the ipsilateral and contralateral testes were significantly increased in groups 2 and 3. While only the hypoxanthine level of group 5 increased significantly, both variables were not significantly different between the ipsilateral testes of groups 1 and 4. CONCLUSIONS: These four treatments damaged both the ipsilateral and contralateral testes. As the lactic acid and hypoxanthine levels within the contralateral testis were greater than in the controls, testicular torsion and vas deferens obstruction seem to share a common pathway (which may be a reflex decrease in contralateral testicular blood flow) for their effects on the contralateral testis. PMID- 10671894 TI - Increased intra-abdominal pressure alters the contractile properties of rabbit bladder. AB - OBJECTIVE: To evaluate the effects of increased intra-abdominal pressure (IAP) on the contractility of the rabbit bladder, as the dynamics of the bladder may be impaired in conditions associated with a high IAP, e.g. constipation and pregnancy. Material and methods The study comprised 22 adult male New Zealand rabbits; six served as the control group, eight had an IAP of 7 cmH2O imposed for 10 days by instilling air into the abdominal cavity and this IAP was maintained for 60 days in a further eight rabbits. After treatment, the rabbits were killed, and the bladders removed and cut into 3 x 12 mm strips. The contractile activity of the muscle strips was then recorded isometrically. Electrical field stimulation (EFS) was applied using a pair of platinum ring electrodes in trains of 3 s duration every 100 s (1 ms, 100 V, 2-100 Hz). Contractile responses to carbachol and isotonic KCl were also evaluated. RESULTS: EFS induced a frequency dependent increase in contractile activity in all bladder strips. Ten days of high IAP resulted in an increased responsiveness to EFS, but high IAP for 60 days reduced the EFS-induced responses to the control levels. Carbachol (10-9-10-3 mol/L) elicited concentration-dependent contractions in all groups. From the concentration-response curves of carbachol, the log EC50 values (the concentration producing half the maximum effect) of the control and 60-day treated animals were comparable, at -6.24 (0.05) and -6.25 (0.04), respectively. However, the log EC50 of the 10 day-treated group was -4.97 (0.08) and significantly (P < 0.01) lower than that of both groups. Isotonic KCl produced contractions in all preparations; these contractions in the control and 60-day treated animals were similar, while the 10 day-treated group had significantly (P < 0.05) higher contraction amplitudes. CONCLUSION: Increased IAP alters the contractile properties of the bladder and its responsiveness to carbachol and KCl. As the intravesical pressure closely reflects the IAP, both should be increased in the present experimental design. PMID- 10671895 TI - Continence and some properties of the urethral striated muscle of male greyhounds. AB - OBJECTIVE: To determine the properties of the striated muscle of the greyhound (dog) urethra and to consider its role in maintaining continence. Materials and methods The thickness of the muscle layers and the muscle types were determined by examining sections stained with haematoxylin and eosin or Masson's trichrome. These factors were correlated with the mechanical and electrical responses of muscle strips to nerve stimulation, and compared with muscle from other breeds of dog and other parts of the animal. RESULTS: The striated muscle formed approximately 70% of the membranous urethra and was predominantly (68%) type IIa muscle (i.e. fast but fatigue-resistant). The mean resting membrane potential was -74 mV; nerve stimulation produced an action potential with a mean amplitude of 97 mV and contraction lasting about 200 ms. All responses were abolished by D tubocurarine. The contractions were well maintained with continuous or intermittent stimulation. The properties were intermediate between those of the anconeus (slow) and the extensor carpi radialis (fast) muscles. CONCLUSIONS: The distribution, fibre type and contractile characteristics would enable the striated urethral muscle to maintain tension for continence at rest and provide additional continence during sprints. PMID- 10671896 TI - The effect of pregnancy and delivery on the function and ultrastructure of the rat bladder and urethra. AB - OBJECTIVE: To examine the effect of pregnancy and delivery on the function and ultrastructure of the bladder and urethra in rats. Material and methods The study comprised six virgin and 18 pregnant rats; both groups underwent cystometry (at the 19th day of gestation, and 2 days and 6 weeks after parturition). Tissues from the bladder and urethra were collected for electron microscopy, western blotting and immunostaining for caveolin-1 and caveolin-3. RESULTS: The bladder capacity was greater and the modified leak-point pressures lower in pregnant and 2-day postpartum rats than in virgin and 6-week postpartum rats. The residual volume was significantly higher in the pregnant group. Electron microscopy showed more sarcolemmal caveolae in the smooth muscle cells of both the bladder and urethra of virgin rats than in the other groups. Lipid droplets and subsarcolemmal mitochondria accumulated in pregnant and 2-day postpartum rats. Caveolin-1 protein was detected in the cytoplasmic membrane of urethra and bladder smooth muscle cells. Caveolin-3 was detected in the membrane of striated muscle in the intrinsic sphincter. Western blotting showed increased caveolin-1 protein expression in the bladder and urethra of 2-day postpartum rats; in contrast, levels of caveolin-1 were lower in pregnant rats than in virgin and 6 week postpartum rats. CONCLUSION: s During pregnancy there was a significant decrease in sarcolemmal caveolae and caveolin-1 in the smooth muscle cells of the rat bladder and urethra. The changes in caveolae and the membrane protein caveolin may play a role in the functional changes associated with pregnancy and after delivery. PMID- 10671897 TI - Homologous bladder augmentation in dog with the bladder acellular matrix graft. AB - OBJECTIVE: To determine the functional potential and antigenicity of the homologous bladder acellular matrix graft (BAMG) in a dog model. MATERIALS AND METHODS: Seven mongrel dogs underwent partial cystectomy (20-50%) and grafting with an equal-sized BAMG; two control animals underwent partial cystectomy (40%) only. The dogs were killed after 30 (one), 120 (one) and 210 days (five dogs). Blood samples were obtained before and at 1, 2, 4, 7, 14, 30, 90 and 210 days after surgery. The dogs underwent cystography, intravenous pyelography and ultrasonography before and after surgery, and on the day they were killed, with cystoscopy carried out just before death. The grafted tissue was assessed using routine and immunohistochemical techniques. RESULTS: All the dogs survived surgery; a complete blood cell count, chemical panel and white blood cell count showed no significant difference between the experimental and control animals. Cystography, cystoscopy and ultrasonography revealed no pathological changes in the upper urinary tract. After 7 months, the mean bladder capacity in the augmented dogs was significantly higher (P = 0.035) than in the controls (264 vs 172 mL). Histological evaluation showed an invasion of all bladder wall components during the first month; at 7 months, the morphological examination showed essentially complete regeneration. CONCLUSION: In this dog model, the potential of the BAMG as a bladder augmentation graft was confirmed, having minimal antigenicity with maximal acceptance. The reconstructed bladder matched the morphological and functional properties of the normal bladder. PMID- 10671898 TI - The treatment of chronic radiation proctitis with hyperbaric oxygen in patients with prostate cancer. PMID- 10671899 TI - Vesicovaginal fistula associated with a vaginal foreign body. PMID- 10671900 TI - Numb-chin syndrome: an unusual presentation of metastatic prostate cancer. PMID- 10671901 TI - Drug trials in children: problems and the way forward. PMID- 10671902 TI - The effect of staggered dosing of sucralfate on oral bioavailability of sparfloxacin. AB - AIMS: To investigate the effect of sucralfate on sparfloxacin absorption when administered concurrently or at strategically spaced dosing times designed to avoid the potential interaction. METHODS: The study was a four-way crossover design where eight healthy Japanese volunteers were randomized to one of four treatment sequences at entry. A 300 mg dose of sparfloxacin was administered alone for treatment A (control). Treatments B, C and D included sucralfate 1.5 g every 12 h for five doses. For treatment B, the fifth dose of sucralfate was administered concurrently with sparfloxacin 300 mg. For treatment C, 300 mg sparfloxacin was given 2 h prior to the fifth dose of sucralfate. Treatment D consisted of sparfloxacin 300 mg given 4 h prior to the fifth dose of sucralfate. Blood and urine samples were collected at predetermined time intervals for 72 h. Sparfloxacin concentrations in plasma and urine and the concentrations of sparfloxacin metabolite in urine were determined by high performance liquid chromatography assays. RESULTS: Sucralfate administrated concurrently with sparfloxacin decreased the mean AUC(0,infinity) of sparfloxacin 2-fold (P<0.001) and the mean Cmax 2.1-fold (P<0.001) compared with sparfloxacin alone. When sucralfate was administrated 2 h after sparfloxacin, the mean AUC(0,infinity) was decreased 1.5-fold (P<0.01) and the mean Cmax 1.4-fold (P<0.01). Sucralfate did not alter the extent of absorption of sparfloxacin when it was given 4 h after the administration of sparfloxacin. The relative bioavailabilities for treatments B, C and D were 0.50 (95% CI: 0.35-0.65), 0.64 (95% CI: 0. 51-0.77), and 0.92 (95% CI: 0.81-1.03), respectively, relative to sparfloxacin alone. The mean percentage of the sparfloxacin dose recovered in urine was significantly lower after sparfloxacin was administered with sucralfate than after sparfloxacin was administered alone or 2 h before the sucralfate dose (P<0.001). Treatments B, C and D were demonstrated to be equivalent to treatment A in the rate of absorption. The tmax, CLr and t1/2 were not significantly affected by sucralfate. The metabolism of sparfloxacin was not altered in the presence of sucralfate. CONCLUSIONS: Oral administration of sucralfate with sparfloxacin or 2 h after sparfloxacin, decreased the extent of sparfloxacin absorption. When both drugs are to be administered together, sucralfate should be administered 4 h after sparfloxacin, allowing thus sufficient time for sparfloxacin absorption prior to the sucralfate dose and thereby minimizing the chance of a significant interaction. PMID- 10671903 TI - Non-linear pharmacokinetics of MDMA ('ecstasy') in humans. AB - AIMS: 3,4-Methylenedioxymethamphetamine (MDMA, commonly called ecstasy) is a synthetic compound increasingly popular as a recreational drug. Little is known about its pharmacology, including its metabolism and pharmacokinetics, in humans in controlled settings. A clinical trial was designed for the evaluation of MDMA pharmacological effects and pharmacokinetics in healthy volunteers. METHODS: A total of 14 subjects were included. In the pilot phase six received MDMA at 50 (n=2), 100 (n=2), and 150 mg (n=2). In the second phase eight received MDMA at both 75 and 125 mg (n=8). Subjects were phenotyped for CYP2D6 activity and were classified as extensive metabolizers for substrates, such as MDMA, whose hepatic metabolism is regulated by this enzyme. Plasma and urine samples were collected throughout the study for the evaluation of MDMA pharmacokinetics. Body fluids were analysed for the determination of MDMA and its main metabolites 3,4 methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxy-methamphetamine (HMMA) and 4 hydroxy-3-methoxy-amphetamine (HMA). RESULTS: As the dose of MDMA administered was increased, volunteers showed rises in MDMA concentrations that did not follow the same proportionality which could be indicative of nonlinearity. In the full range of doses tested the constant recovery of HMMA in the urine combined with the increasing MDMA recovery seems to point towards a saturation or an inhibition of MDMA metabolism (the demethylenation step). These observations are further supported by the fact that urinary clearance was rather constant while nonrenal clearance was dose dependent. CONCLUSIONS: It has previously been postulated that individuals genetically deficient for the hepatic enzyme CYP2D6 (about 10% of the Caucasian people) were at risk of developing acute toxicity at moderate doses of MDMA because the drug would accumulate in the body instead of being metabolized and inactivated. The lack of linearity of MDMA pharmacokinetics (in a window of doses compatible with its recreational use) is a more general phenomenon as it concerns the whole population independent of their CYP2D6 genotype. It implies that relatively small increases in the dose of MDMA ingested are translated to disproportionate rises in MDMA plasma concentrations and hence subjects are more prone to develop acute toxicity. PMID- 10671904 TI - Effects of sibutramine alone and with alcohol on cognitive function in healthy volunteers. AB - AIMS: To investigate the effects of sibutramine in combination with alcohol in a double-blind, randomised, placebo-controlled, four-way crossover study in 20 healthy volunteers. METHODS: On each study day each volunteer received either: sibutramine 20 mg+0.5 g kg-1 alcohol; sibutramine 20 mg+placebo alcohol; placebo capsules+0.5 g kg-1 alcohol; or placebo capsules+placebo alcohol. Alcohol was administered 2 h following ingestion of the study capsules. During each study day, assessments of cognitive performance were made prior to dosing, and at 3, 4.5, 6 and 10 h post dosing. Blood alcohol concentration was estimated using a breath alcometer immediately prior to each cognitive performance test session. Each study day was followed by a minimum 7 day washout period. RESULTS: Alcohol was found to produce statistically significant impairments in tests of attention (maximum impairment to speed of digit vigilance=49 ms) and episodic memory (maximum impairment to speed of word recognition=74 ms). Alcohol also increased body sway (maximum increase 17.4 units) and lowered self rated alertness (maximum decrease 13.6 mm). These effects were produced by an inferred blood alcohol level of 53.2 mg dl-1. Sibutramine was not found to potentiate any of the effects of alcohol. There was a small, yet statistically significant, interaction effect observed on the sensitivity index of the picture recognition task. In this test, the combined effects of sibutramine and alcohol were smaller than the impairments produced by alcohol alone. Sibutramine, when dosed alone, was associated with improved performance on several tasks. Sibutramine improved attention (mean speed of digit vigilance improved by 21 ms), picture recognition speed (improvement at 3=81) and motor control (tracking error at 3 h reduced by 1.58 mm). Also sibutramine improved postural stability (reducing body sway at 3 h by 14.2 units). Adverse events reported were unremarkable and consistent with the known pharmacology of sibutramine and alcohol. CONCLUSIONS: There was little evidence of a clinically relevant interaction of sibutramine with the impairment of cognitive function produced by alcohol in healthy volunteers. The single statistically significant interaction indicated a reduction, rather than a worsening, of alcohol-induced impairment when sibutramine is taken concomitantly. Sibutramine when administered alone is associated with improved performance on several tasks. PMID- 10671905 TI - Pharmacodynamics of milnacipran in young and elderly volunteers. AB - AIMS: To investigate the pharmacodynamics of milnacipran in healthy young and elderly volunteers. METHODS: Randomized double-blind crossover designs were employed and a standardized psychometric battery was administered pre and post dose for both studies. In the first study 10 healthy young volunteers received milnacipran 12.5 mg, 25 mg, 50 mg, 100 mg as a single dose or matched placebo. The test battery was administered at baseline and at 1, 2, 4 and 6 h post dose. The second study compared the effects of milnacipran 75 mg (50 mg+25 mg) per day, amitriptyline 50 mg (25 mg+25 mg) per day and placebo for 3 days' dosing in healthy volunteers aged over 65 years. The test battery was administered at baseline and at 2, 10 and 24 h post dose. The psychometric battery included critical flicker fusion (CFF), choice reaction time (CRT), compensatory tracking (CTT) and tests of short-term memory (STM), subjective sedation (LARS) and subjective sleep (LSEQ). RESULTS: Milnacipran produced no significant dose related effects in the young volunteers. For the elderly, milnacipran significantly (P<0.05) raised CFF scores compared with placebo but had no significant effects on any of the other measures used. Amitriptyline, in contrast, significantly (P<0. 05) lowered CFF threshold, lengthened CRT and increased error on the CTT. On the subjective variables, LARS and LSEQ, amitriptyline increased ratings both of sedation and of difficulty in waking from sleep. CONCLUSIONS: The results showed that milnacipran at single doses of up to 100 mg in healthy young volunteers is free from disruptive effects on cognitive function and psychomotor performance. In addition, milnacipran 75 mg (50+25 mg) appears to be free of negative effects on cognitive function in elderly volunteers, where it seemingly improves performance on CFF. In contrast, the tricyclic antidepressant amitriptyline, used here as a positive internal control, significantly impaired performance in the elderly on the majority of psychometric measures used in this study. This finding not only validated the sensitivity of this current test battery but also indicates the potential behavioural toxicity of amitriptyline in clinical use in the elderly. PMID- 10671906 TI - Comparison of the vasoconstrictor effects of the selective 5-HT1D-receptor agonist L-775,606 with the mixed 5-HT1B/1D-receptor agonist sumatriptan and 5-HT in human isolated coronary artery. AB - AIMS: Vasoconstriction in human coronary artery can be mediated via activation of both 5-HT2 and 5-HT1B-receptors. Coronary vasoconstriction is a rare, but potential adverse effect of the antimigraine drug sumatriptan. In order to investigate the receptor population involved we compared the vasoconstrictor effects of sumatriptan (a mixed 5-HT1B/1D-receptor agonist) with those of L-775, 606 (a selective 5-HT1D-receptor agonist) and 5-HT (the endogenous ligand) in human isolated coronary arteries. METHODS: Coronary arteries were obtained from human hearts removed prior to transplant surgery. Several endothelium denuded ring segments (4 mm in length) were obtained from each artery and mounted for isometric tension recording. Each segment was first exposed to 45 mm KCl and then to 5-HT (1 nm-100 microm ). Concentration-effect curves to L-775,606 (1-(3-(5 (1,2, 4-triazol-4-yl)-1H-indol-3-yl)propyl)-4-(2-(3-fluorophenyl)ethyl)p ipe razine) and sumatriptan were then performed in a consecutive and random manner. The response to repeated application of 5-HT was obtained in separate segments. RESULTS: Twenty-five segments from seven different coronary arteries were studied. Concentration-effect curves were fitted to the data using nonlinear regression analysis. The maximum contraction for L-775,606 was significantly less than that for sumatriptan with Emax values (% relative to 45 mm KCl=100%) of 30.1+/-4.22 and 41.5+/-2.7, respectively. L-775,606 was significantly (30-fold) less potent than sumatriptan in causing contraction compared with sumatriptan (EC50 values were 6.0 microm and 0.2 microm, respectively). For comparison the Emax value for 5-HT was 77.2% and the EC50 value was 0.2 microm. CONCLUSIONS: The selective 5-HT1D-receptor agonist L-775,606 has less propensity towards vasoconstriction in human isolated coronary artery (endothelium-denuded) than was mixed 5-HT1B/1D-receptor agonist sumatriptan. The contractions produced were at concentrations where L-775,606 would be expected to occupy 5-HT1B-receptors. PMID- 10671907 TI - Pharmacokinetics and pharmacodynamics of R- and S-gallopamil during multiple dosing. AB - AIMS: Using a stable isotope technique we investigated the pharmacokinetics and pharmacodynamics of gallopamil after administration of 50 mg pseudoracemic gallopamil every 12 h for 7 doses (72 h). METHODS: Six male healthy volunteers were studied. After the seventh dose the pharmacokinetics and pharmacodynamics were assessed. Serum levels of gallopamil were measured by gas chromatography/mass spectrometry. Effects of gallopamil were measured by ECG recording. RESULTS: The apparent oral clearances (R: 4.8 l min-1 (95% CI: 2.9 6.8); S: 5.5 l min-1 (95% CI: 2.5-8.5)) and half-lives (R: 6.2 h; S: 7.2 h) of R- and S-gallopamil were similar (P >0.05). The serum protein binding (fu R: 0.035 (95% CI: 0.026-0. 045); S: 0.051 (95% CI: 0.033-0.069)) and the renal elimination (% of dose R: 0.49%; S: 0.71%) were enantioselective. Gallopamil had a potent effect on the PR interval (% prolongation 35.7% (95% CI: 14. 0-57.3)). No changes in other electrocardiographic or cardiovascular parameters were observed. CONCLUSIONS: The pharmacokinetics and bioavailability of the racemic drug gallopamil are not stereoselective at steady-state and are therefore not substantially altered compared with the single dose administration of gallopamil. PMID- 10671908 TI - Formation of omeprazole sulphone but not 5-hydroxyomeprazole is inhibited by grapefruit juice. AB - AIMS: To determine the effect of grapefruit juice on omeprazole metabolism in vivo. METHODS: This was a randomized crossover study with a 2 week washout period. Omeprazole (20 mg) was taken orally by 13 healthy volunteers after an overnight fast with either grapefruit juice or water. Serial blood samples were obtained over 12 h and standardized meals were served 3 and 10 h after the administration of omeprazole. Plasma concentrations of omeprazole and its major metabolites, 5-hydroxyomeprazole and omeprazole sulphone, were determined by high performance liquid chromatography (h.p.l.c.). RESULTS: Mean area under the plasma concentration vs time curve (AUC) between 0 and 12 h for omeprazole sulphone was approximately 20% lower (P<0.01) in the group receiving grapefruit juice. There was no significant difference in the mean AUC of 5-hydroxyomeprazole or omeprazole. The AUC ratio of omeprazole sulphone to omeprazole, an index of CYP3A4 activity, was decreased by 33% (P<0.001) after administration of grapefruit juice whereas the AUC ratio of 5-hydroxyomeprazole to omeprazole, an index of CYP2C19 activity, did not differ between the two phases of the study. Although the time to peak concentration (tmax ) and terminal half-life (t1/2,z) of omeprazole and its two main metabolites were not altered, the peak concentration (Cmax ) of omeprazole sulphone was significantly reduced after administration of grapefruit juice. CONCLUSION: Administration of grapefruit juice decreased the formation of omeprazole sulphone but not 5-hydroxyomeprazole. These results indicate that activities of CYP3A4, but not of CYP2C19, are inhibited by the simultaneous administration of grapefruit juice. PMID- 10671909 TI - Phenotypic polymorphism and gender-related differences of CYP1A2 activity in a Chinese population. AB - AIMS: To investigate the distribution characteristics of CYP1A2 in a Chinese population, and to examine gender-related differences in CYP1A2 activity. METHODS: Two hundred and twenty-nine healthy subjects, 120 men and 109 women, were enrolled in this study. CYP1A2 activity was measured by plasma paraxanthine/caffeine (1,7X/1,3,7X) ratio 6 h after administration of 300 mg caffeine. The concentrations of paraxanthine and caffeine in plasma were detected by h.p.l.c. RESULTS: A 16-fold variation of CYP1A2 activity (range 0. 09 to 1.46) was shown in this study. The coefficient of variation (CV %) of CYP1A2 activity was 62.9%. Non-normal distribution of CYP1A2 activity was indicated by the Shapiro-Wilk test (P<0.001). Probit plots of CYP1A2 activity revealed a bimodal distribution with breakpoint of 1,7X/1,3,7X ratio of 0.12. The percentage of poor metabolizers (PMs) was 5.24% (95% CI: 2.35% approximately 8.13%) in this Chinese population. Residual analysis of the data also supported bimodality (P<0.01). The CYP1A2 activity of men was higher than that of women (median: 0.33 vs 0.23, P<0.001). A probit plot of CYP1A2 activity in men was shifted to the left compared with that in women. Based on phenotype, the gender-related difference was observed in extensive metabolizers (EMs) (P<0.001), but not in PMs (P >0.1). In addition, there was no sex-related difference in the incidence of PMs (P >0.1). CONCLUSIONS: There is a phenotypic polymorphism in CYP1A2 activity in this Chinese population, and CYP1A2 activity is higher in men than that in women. PMID- 10671910 TI - Stereoselective glucuronidation of formoterol by human liver microsomes. AB - AIMS: Formoterol is a beta2-adrenoceptor agonist marketed as a racemic mixture of the active (R; R)- and inactive (S; S)-enantiomers (rac-formoterol). The drug produces prolonged bronchodilation by inhalation but there is significant interpatient variability in duration of effect. Previous work has shown that in humans formoterol is metabolized by conjugation with glucuronic acid but little is known about the stereoselectivity of this reaction. The aim of the present study was to investigate the glucuronidation of formoterol enantiomers in vitro by human liver microsomes. METHODS: The kinetics of formation of formoterol glucuronides during incubation of racemate and of single formoterol enantiomers with human liver microsomes (n=9) was characterized by chiral h.p.l.c. assay. RESULTS: The kinetics of glucuronidation of the two formoterol enantiomers obeyed the Michaelis-Menten equation. Glucuronidation of formoterol was stereoselective and occurred more than two times faster for (S; S)-formoterol than for (R; R) formoterol. In incubations with single formoterol enantiomers, the median (n=9) Km values for (R; R)-glucuronide and (S; S)-glucuronide were 827.6 and 840.4 microm, respectively, and the median V max values were 2625 and 4304 pmol min-1 mg-1, respectively. Corresponding values determined in incubations with rac formoterol were 357.2 and 312.1 microm and 1435 and 2086 pmol min-1 mg-1 for (R; R)- and (S; S)-glucuronide, respectively. Interindividual variation was large with the ratio of V max/Km (S; S/R; R) ranging from 0.57 to 6.90 for incubations with rac-formoterol. CONCLUSIONS: Our study demonstrates that glucuronidation of formoterol by human liver microsomes is stereoselective and subject to high interindividual variability. These findings suggest that clearance of formoterol in humans is subject to variable stereoselectivity which could explain the variation in duration of bronchodilation produced by inhaled formoterol in patients with asthma. PMID- 10671911 TI - Epidemiology of drug exposure and adverse drug reactions in two swiss departments of internal medicine. AB - AIMS: To explore drug exposure, frequency of adverse drug reactions (ADRs), types of ADRs, predisposing risk factors and ADR-related excess hospital stay in medical inpatients. METHODS: Structured data regarding patient characteristics, 'events' (symptoms, laboratory results), diagnoses (ICD10) and drug therapy were collected using a computer-supported data entry system and an interface for data retrieval from electronic patient records. ADR data were collected by 'event monitoring' to minimize possible bias by the drug monitor. The causality of each event was assessed in relation to disease(s) and drug therapy. RESULTS: The analysis included 4331 (100%) hospitalizations. The median observation period was 8 days. The median number of different drugs administered per patient and day was 6 and varied between 4 (Q1 ) and 9 (Q3 ) different drugs in 50% of all hospital days. In 41% of all hospitalizations at least one disease-unrelated event could be possibly attributed to drug therapy. Clinically relevant ADRs occurred in 11% of all hospitalizations. In 3.3% of all hospitalizations ADRs were the cause of hospital admission. The incidence of possibly ADR-related deaths was 1.4. Factors predisposing for clinically relevant ADRs were female gender and polypharmacy. ADR-related excess hospital stay accounted for 8. 6% of hospital days. CONCLUSIONS: These data demonstrate the feasibility of the developed 'event monitoring' system for quantitative analysis of ADRs in medical inpatients. With increasing numbers of recorded patients the pharmacoepidemiological database provides a valuable tool to study specific questions regarding drug efficacy and safety in hospitalized patients. PMID- 10671912 TI - Modification of general practitioner prescribing of antibiotics by use of a therapeutics adviser (academic detailer). AB - AIMS: This was a pilot study of the use of a clinical pharmacist as a therapeutics adviser (academic detailer) to modify antibiotic prescribing by general practitioners. METHODS: Following a visit by the adviser (March-May), 112 general practitioners were recruited and randomised to control or active groups. A panel of experts prepared a best practice chart of recommended drugs for upper and lower respiratory tract infections, otitis media and urinary tract infections. The adviser made a 10-15 min visit to each prescriber in the active group (June-July), gave them the chart and discussed its recommendations briefly. Doctors in the control group were not visited nor given the chart. Prescription numbers for all prescribers were obtained from the Commonwealth Health Insurance Commission for the pre(March-May) and postdetailing (August-September) periods using a three month lag time for data collection. Data for total numbers of prescriptions and for selected individual antibiotics used in these two periods were analysed using nonparametric statistics. RESULTS: Prescribing patterns were similar for the control and active groups in the predetailing period. For both groups, there were significant (P<0.03) increases (45% for control and 40% for active) in total number of antibiotic prescriptions in the post compared with the predetailing period. This trend was anticipated on the basis of the winter seasonal increase in respiratory infections. In line with the chart recommendations for first-line treatment, doctors in the active group prescribed significantly more amoxycillin (P<0.02) and doxycycline (P<0.001) in the post vs predetailing periods. By contrast, doctors in the control group prescribed significantly more cefaclor (P<0.03) and roxithromycin (P<0.03), drugs that were not recommended. The total cost of antibiotics prescribed by doctors in the control group increased by 48% ($37 150) from the preto postdetailing periods. In the same time period, the costs for the active group increased by only 35% ($21 020). CONCLUSIONS: We conclude that the academic detailing process was successful in modifying prescribing patterns and that it also decreased prescription numbers and costs. Application of the scheme on a nationwide basis could not only improve prescriber choice of the most appropriate antibiotic but also result in a significant saving of health care dollars. PMID- 10671913 TI - Sleeping with the enemy? A randomized controlled trial of a collaborative health authority/industry intervention to influence prescribing practice. AB - AIMS: To evaluate the effectiveness of a health authority/pharmaceutical company collaborative intervention to influence the choice of proton pump inhibitors METHODS: Randomized controlled trial, with general practices forming the unit of allocation and analysis. RESULTS: Constructive working relationships were achieved with five of six pharmaceutical companies involved. One hundred and two out of 140 practitioners in intervention group practices received at least one visit from an industry representative. There were no reports of representatives operating outside their agreed remit. Prescribing in both the intervention and control group moved towards that recommended by the guidelines but there was no difference between the groups in either the proportion of prescriptions in line with the guidelines or the overall cost. CONCLUSIONS: Health authorities can achieve professional working relationships with the pharmaceutical industry although no changes in practice attributable to the intervention are achieved. Further work is required to develop effective means to influence prescribing in line with independent guidelines especially in the context of the development of Primary Care Groups. PMID- 10671915 TI - Neuroendocrine tumours of the pancreas. PMID- 10671914 TI - Inhibition of debrisoquine hydroxylation with quinidine in subjects with three or more functional CYP2D6 genes. AB - AIMS: To study whether the CYP2D6 capacity in ultrarapid metabolizers of debrisoquine due to duplication/multiduplication of a functional CYP2D6 gene, can be 'normalised' by low doses of the CYP2D6 inhibitor quinidine and whether this is dose-dependent. METHODS: Five ultrarapid metabolizers of debrisoquine with 3, 4 or 13 functional CYP2D6 genes were given single oral doses of 5, 10, 20, 40, 80 and 160 mg quinidine. Four hours after quinidine intake, 10 mg debrisoquine was given. Urine was collected for 6 h after debrisoquine administration. Debrisoquine and its 4-hydroxymetabolite were analysed by h.p.l.c. and the debrisoquine metabolic ratio (MR) was calculated. RESULTS: Without quinidine the MR in the ultrarapid metabolizers ranged between 0.01 and 0.07. A dose-effect relationship could be established for quinidine with regard to the inhibitory effect on CYP2D6 activity. To reach an MR of 1-2, subjects with 3 or 4 functional genes required a quinidine dose of about 40 mg, while the sister and brother with 13 functional genes required about 80 mg quinidine. After 160 mg quinidine, the MRs, in the subjects with 3, 3, 4, 13 and 13 functional genes, were 12.6, 10.1, 9.2, 2.4 and 2.2, respectively. CONCLUSIONS: A dose-effect relationship could be established for quinidine inhibition of CYP2D6 in ultrarapid metabolizers. The clinical use of low doses of quinidine as an inhibitor of CYP2D6 might be considered in ultrarapid metabolizers taking CYP2D6 metabolized drugs rather than giving increased doses of the drug. Normalizing the metabolic capacity of CYP2D6, by giving a low dose of quinidine, may solve the problem of 'treatment resistance' caused by ultrarapid metabolism. PMID- 10671916 TI - What should surgeons write? PMID- 10671917 TI - COPE: Committee on Publication Ethics. Guidelines on good publication practice. PMID- 10671918 TI - The 'failed' embolectomy. PMID- 10671919 TI - Xenotransplantation. AB - BACKGROUND: The success of clinical transplantation has led to a large discrepancy between donor organ availability and demand; considerable pressure exists to develop an alternative source of organs. The use of animal organs for donation is a possible solution that is not yet clinically applicable. METHODS AND RESULTS: A literature review was performed based on a Medline search to find articles on xenotransplantation. Keywords included hyperacute, acute vascular, xenograft rejection combined with concordant and discordant. Additional references cited in these articles from journals not included in Medline were obtained from the British Library. Limited information on unpublished, preliminary work has been included from sources known to the authors, based on their research work in the field. One hundred and forty-six references and four personal communications have been included in this review article. CONCLUSION: A greater understanding of the pathogenesis of xenograft rejection is developing rapidly. Strategies to abrogate hyperacute rejection have proved successful, but control of antibody-driven acute vascular rejection has not yet been achieved. The safety and viability of xenotransplantation as a therapeutic modality are still unproven. PMID- 10671920 TI - Genetic predisposition to breast cancer: a surgical perspective. AB - BACKGROUND: Molecular alterations in proto-oncogenes, tumour suppressor genes, and genes that function in DNA damage recognition and repair are considered to be hallmarks of a carcinogenic process, including breast carcinogenesis. METHODS: A computer-assisted search of the English literature (Medline database, 1990-1999) was performed, followed by a manual search of the reference list of pertinent articles retrieved. RESULTS: Hereditary breast cancer accounts for 5-10 per cent of all breast cancer cases. About 90 per cent of hereditary breast cancers involve mutation of the BRCA1 and/or BRCA2 genes. Other cancer-related genes (including myc, c-erbB2, Tsg101 and Mdgi) are involved in breast carcinogenesis, but they do not give rise to familial breast cancer syndromes. Risk estimation is the most important clinical implication. Management options for the high-risk mutation carriers include cancer surveillance and preventive strategies (prophylactic surgery or chemoprevention). CONCLUSION: Despite inadequate knowledge about the genetic predisposition to breast cancer and its clinical implications, the demand for genetic testing is likely to expand rapidly. In addition to risk estimation, cancer surveillance and preventive strategies, gene therapy offers a new and theoretically attractive approach to breast cancer management. PMID- 10671921 TI - Management of the axilla in operable breast cancer treated by breast conservation: a randomized clinical trial. Edinburgh Breast Unit. AB - BACKGROUND: In the treatment of operable breast cancer by breast conservation, the extent of axillary dissection, the need for radiotherapy to the axilla and the morbidity associated with these procedures have not been assessed adequately. METHODS: Patients with operable breast cancer were randomized to have level III axillary node clearance (232 patients) or axillary node sample (234 patients). Radiotherapy to the axilla was given selectively. Radiotherapy was not given to those who had an axillary clearance. In the early part of the study all patients who had node sample were treated by radiotherapy (54 patients); subsequently this was modified to include only those who were node positive. The morbidity to the shoulder and arm was assessed before and after operation by measuring upper limb volume and circumference, and combined glenohumeral and scapular movement and muscle power. RESULTS: Comparing the two surgical policies, no difference was found in local (axillary clearance 14 versus sample 15), axillary (eight versus seven) or distant (29 versus 29) recurrence. There was no statistically significant difference in 5-year survival rate (clearance 82.1 versus sample 88.6 per cent). Morbidity was least in those who had a node sample and no radiotherapy to the axilla. Radiotherapy to the axilla in patients who had a node sample resulted in a significant reduction in range of movement of the shoulder, e.g. mean(s.e.) 2.2(0.6) cm reduction in lateral rotation at 3 years. Surgical axillary clearance was associated with significant lymphoedema of the upper limb, e.g. 4.1(0.7) per cent increase in arm volume at 3 years. CONCLUSION: A selective policy for the management of the axilla is associated with no increase in axillary recurrence or mortality rate compared with routine axillary node clearance. Patients who are node negative after axillary sample can avoid radiotherapy or axillary clearance. PMID- 10671922 TI - Mutations of the cationic trypsinogen gene in patients with hereditary pancreatitis. AB - BACKGROUND: Hereditary pancreatitis has been shown to be caused by one of two mutations (R117H and N21I) of the cationic trypsinogen gene (PRSS1). Families with hereditary pancreatitis in the north of England were investigated for these mutations. The clinical features associated with each mutation were compared. METHODS: In individuals from nine families with hereditary pancreatitis, DNA was screened for the R117H and N21I mutations. All five exons of the cationic trypsinogen gene were also sequenced to search for additional mutations. Haplotype analysis was carried out to identify common ancestors. Clinical data were collected. RESULTS: The R117H mutation was identified in three families and N21I in a further five. The R117H mutation was associated with a more severe phenotype than N21I in terms of mean(s.d.) age of onset of symptoms (8.4(7.2) versus 16. 5(7.1) years; P = 0.007) and requirement for surgical intervention (eight of 12 versus four of 17 patients respectively; P = 0.029). Haplotype analysis suggested that each mutation had arisen more than once. CONCLUSION: Two mutations in the cationic trypsinogen gene cause hereditary pancreatitis in eight of nine families originating in this region. The R117H mutation is associated with a more severe form of the disease in terms of age at onset of symptoms and requirement for surgical intervention. PMID- 10671923 TI - Role of relaparoscopy in the management of minor bile leakage after laparoscopic cholecystectomy. AB - BACKGROUND: Bile leakage in the absence of major ductal injury may occur from the liver bed or from the cystic duct remnant after cholecystectomy. The early limitations of minimally invasive surgery led to reliance on endoscopic methods to manage this complication. However, repeat laparoscopy permits drainage of the bile collection and direct control of the site of leakage in selected situations. METHODS: Details of 15 patients with bile leakage after laparoscopic cholecystectomy were recorded prospectively and are reviewed. RESULTS: Postoperative bile leakage occurred after 15 (0.8 per cent) of 1779 laparoscopic cholecystectomies. Two patients with bile in drainage fluid had spontaneous resolution. Ten patients with a subvesical duct leak had repeat laparoscopy. The leak was successfully controlled by suturing in eight patients, and by a laparoscopically placed drain in two. One patient required a subsequent laparotomy for a loculated pelvic collection. Three patients had cystic duct stump leakage. This was managed successfully by laparoscopy in one case but required endoscopic management in two. CONCLUSION: Laparoscopy is useful in the management of minor bile leaks after laparoscopic cholecystectomy. Selection of appropriate patients relies on a characteristic clinical presentation after an otherwise uncomplicated cholecystectomy. PMID- 10671924 TI - Role of biliary scintiscan in predicting the need for cholangiography. AB - BACKGROUND: Currently used predictors for bile duct calculi in patients undergoing cholecystectomy have low specificity resulting in unnecessary cholangiograms being performed. The role of biliary scintiscan in predicting the presence of bile duct calculi was assessed. METHODS: Seventy-five patients with symptomatic gallstone disease were studied prospectively regard- ing the value of a history of jaundice or acute pancreatitis, raised serum bilirubin and serum alkaline phosphatase levels, and visualization of stones or presence of dilated bile ducts on ultrasonography (standard criteria) in detecting bile duct calculi. Results of biliary scintiscan were evaluated against a combination of standard criteria. The 'gold standard' for evaluation was endoscopic or peroperative cholangiography. RESULTS: Biliary scintiscan had a higher sensitivity and specificity (93 and 94 per cent) than a combination of the above standard and modified predictors for biliary calculi (89 and 71 per cent). A combination of ultrasonography and selective use of scintiscan, in the absence of bile duct dilatation only, had higher values (96 and 98 per cent). CONCLUSION: A combination of ultrasonography and biliary scintiscan can accurately predict bile duct calculi and could be used as a guide for selective cholangiography. PMID- 10671925 TI - Spontaneous carotid artery aneurysms. AB - BACKGROUND: Spontaneous carotid artery aneurysms are infrequently reported, and are almost always non-atherosclerotic. METHODS: The records of 29 patients with a spontaneous carotid aneurysm treated in an academic vascular unit between 1990 and 1998 were reviewed. RESULTS: All 29 patients were black South Africans; three had bilateral aneurysms. There were 24 men and five women, of mean age 35 (range 13-62) years. Some 25 aneurysms involved the common carotid artery, 12 of which affected the bifurcation, and seven were located in the internal carotid artery. Twenty-five aneurysms were managed surgically, four of which were ligated owing to sepsis. Histo- logical evaluation showed human immunodeficiency virus-related arteritis in four, tuberculous aneurysms in ten, Takayasu's arteritis in two, atherosclerosis in three and non-specific chronic inflammation in four patients. Microbiological examination was negative in all but one patient who had Salmonella sp. cultured. Outcome was generally favourable, but one patient died from massive hemispheric infarction. There were no other new neurological deficits. CONCLUSION: Carotid aneurysms pose a considerable surgical challenge but are amenable to operative intervention with good result. Ligation appears to be well tolerated in this group of predominantly non-atherosclerotic aneurysms. PMID- 10671926 TI - Influence of gender on outcome from ruptured abdominal aortic aneurysm. AB - BACKGROUND: The aim of the present study was to compare outcomes following ruptured abdominal aortic aneurysm (AAA) in men and women. METHODS: Overall mortality from ruptured AAA was compared in men and women using the Western Australia Health Services Research Database. The linked chains of de-identified hospital morbidity and death records were selected using the ICD-9-CM (International Classification of Diseases - Clinical Modification) diagnostic and procedure codes pertaining to AAA. Cases were divided into three groups for analysis: patients who died without admission to hospital, those admitted to hospital with a ruptured AAA but who did not undergo operation, and patients who underwent operation for ruptured AAA. RESULTS: Ruptured AAA occurred in 648 men and 225 women over the age of 55 years during the decade 1985-1994. Only 50 per cent of women, compared with 59 per cent of men, were admitted to hospital. Of those admitted to hospital only 37 per cent of women underwent operation, compared with 63 per cent of men. The overall mortality rate from ruptured AAA was 90 per cent in women and 76 per cent in men (chi2 = 50.34, 1 d.f., P < 0.0001). Although women were, on average, 6 years older than men, this unfavourable pattern occurred across all age groups. CONCLUSION: Women with a ruptured AAA are more likely to die than men. More research is required to identify the causes of this sex difference. PMID- 10671927 TI - Quantifying the risks of hypertension, age, sex and smoking in patients with abdominal aortic aneurysm. AB - BACKGROUND: The prevalence of abdominal aortic aneurysm (AAA) in a community based sample of men and women aged 65-79 years was correlated with known risk factors. In addition, the effect of high blood pressure and the use of antihypertensive medication on growth of AAAs were studied. METHODS: Aortic diameter was assessed by ultrasonography and data on risk factors were collected by self-administered questionnaire for 5356 men and women as part of a randomized controlled trial. RESULTS: Current hypertension increased the risk of having an aortic aneurysm by 30-40 per cent while use of antihypertensive medication increased the risk by 70-80 per cent, adjusting for current blood pressure. There was no clear relationship between hypertension and growth rates of existing aneurysms in this study, although these results were largely from data on small aneurysms. Men were nearly six times more likely to develop an AAA than women; the risk increased by 40 per cent every 5 years after the age of 65 years. Smoking was an independent risk factor for AAA, with level of exposure being more significant than duration. CONCLUSION: Male sex, smoking and hypertension are strong risk factors for the development of AAA. In this study hypertension did not significantly increase the growth rate of existing aneurysms. Smoking remains the most important avoidable risk factor for AAA. The analyses presented here suggest that selection for screening, other than by age and sex, is not worthwhile. PMID- 10671928 TI - Diaphragmatic dysfunction secondary to experimental lower torso ischaemia reperfusion injury is attenuated by thermal preconditioning. AB - BACKGROUND: Preconditioning describes the process whereby tissue exposure to a subcritical stress confers protection from subsequent injuries. This study assessed diaphragmatic muscle function after lower torso ischaemia-reperfusion (IR) and the role of thermal preconditioning in attenuation of this injury. METHODS: Sprague-Dawley rats were randomized into three groups (24 per group): a control group, an IR group that had aortic cross-clamping for 1 h followed by reperfusion, and a third group that received thermal preconditioning 18 h before IR. Diaphragmatic function was assessed at 24 h, 48 h and 7 days. RESULTS: IR resulted in significant diaphragmatic twitch and tetanic dysfunction compared with control muscle. Thermal preconditioning significantly attenuated this injury (P < 0.05). Mean(s.e.m.) muscle twitch and tetanic forces in the IR group were 204.9(17.2) and 282.7(19.2) g respectively at 24 h. Corresponding twitch and tetanic forces in preconditioned muscle were 270.4(25.1) and 552.0(35.2) g. CONCLUSION: This study demonstrated that systemic IR injury produced a respiratory muscle mechanical dysfunction that was attenuated by thermal preconditioning, at 24 h, 48 h and 7 days. Preconditioning may have a role in clinical practice, particularly before elective surgery. PMID- 10671929 TI - Bladder and sexual dysfunction after mesorectal excision for rectal cancer. AB - BACKGROUND: Urinary and sexual dysfunction are recognized complications of rectal excision for cancer. The aim of this study was to examine the frequency of such complications after mesorectal excision, shortly after this method was introduced. METHODS: Spontaneous flowmetry, residual volume of urine measurement and urodynamic examination, including cystometry and simultaneous detrusor pressure and urinary flow recording, was carried out before and 3 months after curative rectal excision. Urinary symptoms and sexual function were evaluated by means of questionnaires before and after operation. Each patient served as his or her own control. RESULTS: Forty-nine consecutive patients, 39 of whom had a total mesorectal excision (TME) and ten a partial mesorectal excision, were examined before surgery and 35 again after operation. In two patients, a weak detrusor was detected before operation. Two patients developed signs of bladder denervation after operation. Transitory moderate urinary incontinence appeared in four other women. Six of 24 men reported some reduction in erectile function and one became impotent. Two men reported retrograde ejaculation. All the complications were seen in the TME group. CONCLUSION: Mesorectal excision for rectal cancer resulted in a low frequency of serious bladder and sexual dysfunction. PMID- 10671930 TI - Prolonged ambulatory recording of antroduodenal motility in slow-transit constipation. AB - BACKGROUND: Slow-transit constipation may be part of a pan-enteric motor disorder. To test this hypothesis 24-h ambulatory antroduodenal manometry was performed and orocaecal transit time determined in patients with slow-transit constipation and in healthy controls. METHODS: Antroduodenal motility was recorded with a five-channel solid-state catheter. Postprandial motility was recorded after consumption of two standardized test meals and interdigestive motility was recorded nocturnally. Manometry tracings were analysed for quantitative and qualitative abnormalities. Orocaecal transit time was determined by means of the lactulose hydrogen breath test. RESULTS: Postprandial motility was no different between patients and controls. However, some minor changes of interdigestive motility were observed. The proportion of phase II activity of the nocturnal cycles of the interdigestive migrating motor complex was increased in patients while phase I activity was decreased. The total number of observed phase III fronts was no different in patients and controls, although the number of phase III fronts with antral onset was decreased. Furthermore, the amplitude of phase III activity of duodenal onset was also decreased. Specific motor abnormalities such as retrograde propagation of phase III fronts were more frequent in patients. Orocaecal transit time was delayed in patients. CONCLUSION: In patients with slow-transit constipation, orocaecal transit time is delayed but antro- duodenal motility is generally well preserved with only minor alterations. Presented as a poster to the Digestive Disease Week meeting in New Orleans, Louisiana, USA, May 1998, and published in abstract form as Gastroenterology 1998; 114: A820 PMID- 10671931 TI - Evaluation of Delorme's procedure as a treatment for full-thickness rectal prolapse. AB - BACKGROUND: Delorme's procedure is a well tolerated perineal operation for full thickness rectal prolapse. However, prolapse recurrence is common and reported recurrence rates vary widely. This study attempted to standardize outcome assessment for recurrence following primary and subsequent Delorme's operations. Patient and operative factors were analysed to identify any that might improve patient selection. METHODS: Some 101 primary and 17 secondary Delorme's procedures were carried out on 113 consecutive patients presenting with rectal prolapse, who were followed for a minimum of 12 months, unless death or recurrent prolapse intervened. The rate of prolapse recurrence was calculated using the Kaplan-Meier method of analysis. Patient age, sex, grade of incontinence, presence of diverticular disease, length of mucosal resection and position in the operative series were analysed to identify factors affecting recurrence. RESULTS: The predicted recurrence-free period for 50 per cent of patients undergoing primary and secondary Delorme's procedures was 91 (95 per cent confidence interval 77-105) and 27 (15-39) months respectively. None of the patient or operative factors analysed was related to recurrent prolapse. CONCLUSION: Delorme's procedure is a simple operation with satisfactory functional results which can be considered in all patients of all ages. However, high recurrence rates for primary and repeat operations should be explained to patients when planning their surgical management. PMID- 10671932 TI - Lymphocyte activation after non-thermal trauma. AB - BACKGROUND: Severe injury causes immunological changes that may contribute to a poor outcome. Longitudinal characterization of lymphocyte response patterns may provide further insight into the basis of these immunological alterations. METHODS: Venous blood obtained seven times over 2 weeks from 61 patients with injury severity scores above 20 was assessed for lymphocyte phenotypic and activation markers together with serum levels of interleukin (IL) 2, IL-4, soluble IL-2 receptor (sIL-2R), soluble CD4 (sCD4), soluble CD8 (sCD8) and interferon gamma. RESULTS: Severe injury was associated with profound changes in the phenotypic and activation profile of circulating lymphocytes. Activation was indicated by increased numbers of T cells expressing CD25, CD69 and CD71, and raised serum levels of IL-2, sIL-2R, sCD4 and sCD8. Relatively higher levels of sIL-2R and sCD4 were found in patients with sepsis syndrome. CONCLUSION: Polytrauma is associated with dramatic alterations in the phenotypic and activation profile of circulating lymphocytes which are generally independent of clinical course. In contrast, several lymphocyte soluble factors, including sCD4 and sIL-2R, paralleled the clinical course. These data provide new insight into lymphocyte responses after injury and suggest that further assessment of soluble factors as clinical correlates, including those related to lymphocyte activation or generalized inflammation, may be warranted. PMID- 10671933 TI - Acrylate yellow filters in operating lights protect against photosensitization tissue damage. AB - BACKGROUND: Photosensitized patients are exposed to bright lights when undergoing intraoperative photodynamic therapy or fluorescence measurements. Acrylate yellow filters might reduce unwanted tissue damage. METHODS: To investigate the protective value of these filters, the spectral power distribution of the operating lights and light energy densities with and without an acrylate yellow filter were measured. Subsequently the effects of light exposure on the survival of a human hepatocellular carcinoma cell line and the photodamage induced in pig tissues after the administration of 5-aminolaevulinic acid were also studied. RESULTS: The light energy density in the ultraviolet and blue region of the light spectrum emitted by the operating light was reduced up to 50 per cent by the acrylate yellow filter. The survival of photosensitized cells was longer and photodamage induced in pig tissues was less when exposed to filtered light. CONCLUSION: Photodamage induced by operating lights can be reduced by filtering out ultraviolet and blue light by means of acrylate yellow filters. PMID- 10671934 TI - Recurrence following curative resection for gastric carcinoma. AB - BACKGROUND: The diagnosis and treatment of recurrent gastric cancer remains difficult. The aim of this study was to determine the risk factors for recurrence of gastric cancer and the prognosis for these patients. METHODS: Of 2328 patients who underwent curative resection for gastric cancer from 1987 to 1995, 508 whose recurrence was confirmed by clinical examination or reoperation were studied retrospectively. The risk factors that determined the recurrence patterns and timing were investigated by univariate and multivariate analysis. RESULTS: The mean time to recurrence was 21.8 months and peritoneal recurrence was the most frequent (45.9 per cent). Logistic regression analysis showed that serosal invasion and lymph node metastasis were risk factors for all recurrence patterns and early recurrence (at 24 months or less). In addition, independent risk factors involved in each recurrence pattern included younger age, infiltrative or diffuse type, undifferentiated tumour and total gastrectomy for peritoneal recurrence; older age and larger tumour size for disseminated, haematogenous recurrence; and older age, larger tumour size, infiltrative or diffuse type, proximally located tumour and subtotal gastrectomy for locoregional recurrence. Other risk factors for early recurrence were infiltrative or diffuse type and total gastrectomy. Reoperation for cure was possible in only 19 patients and the mean survival time after conservative treatment or palliative operation was less than 12 months. CONCLUSION: The risk factors for each recurrence pattern and timing of gastric cancer can be predicted by the clinicopathological features of the primary tumour. Since the results of treatment remain dismal, studies of perioperative adjuvant therapy in an attempt to reduce recurrence are warranted. PMID- 10671935 TI - Surgical treatment for recurrent gastro-oesophageal reflux disease after failed antireflux surgery. AB - BACKGROUND: Recurrent or persistent symptoms occur in 10-15 per cent of patients after antireflux surgery. Failure of surgery is not uniform in its presentation. The cause of failure is not easily detected and even harder to treat. Different approaches have been proposed and few reports are available on the objective and subjective outcome of reoperation. METHODS: This study focuses on 30 patients (16 men and 14 women; age range 20-69 years) with recurrent symptomatic gastro oesophageal reflux disease (GORD) resistant to medical treatment. In all patients reoperation was by the Belsey Mark IV antireflux operation. A clinical history, endoscopy and oesophageal manometry were obtained in all patients, and 24-h pH monitoring was performed in 27 of 30 before and in most patients after the Belsey procedure. RESULTS: Symptomatic improvement was reported in 24 of 30 patients. Oesophagitis (present before operation in 19 patients) was cured or remained absent in 24 of 30 patients, stabilized in one, improved in four and deteriorated in one. Relief of symptoms combined with absence of oesophagitis was obtained in 21 of 30 patients, with concomitant normalization of the 24-h pH profile in 11 of 22 patients. The median basal lower oesophageal sphincter (LOS) pressure increased significantly from 6. 9 to 9.0 mmHg (P < 0.01). Redo surgery had no effect on oesophageal body motility. CONCLUSION: Reoperation performed for documented recurrent GORD had a good and lasting effect on symptoms, on oesophagitis (both in 24 of 30 patients) and on the combination of both (21 of 30). In these patients reoperation increased basal LOS pressure and decreased reflux time. Overall, the results approximate to those of primary operation. PMID- 10671936 TI - Screening children at risk of developing inherited endocrine neoplasia syndromes. PMID- 10671937 TI - New causes of Cushing's syndrome: abnormal adrenal regulation of cortisol production. PMID- 10671938 TI - CTLA4 variants in type 1 diabetes: some stirrups serve better backing endocrine autoimmunity. PMID- 10671939 TI - Application of a disease-specific, quality-of-life measure (QoL-AGHDA) in growth hormone-deficient adults and a random population sample in sweden: validation of the measure by rasch analysis. PMID- 10671940 TI - Application of a disease-specific, quality-of-life measure (QoL-AGHDA) in growth hormone-deficient adults and a random population sample in Sweden: validation of the measure by rasch analysis. AB - OBJECTIVE: Growth hormone deficiency (GHD) in adults has been associated with impaired health status and quality of life (QoL) in several studies using generic measures, and in a few studies using recently developed disease-specific measures. Theoretically, disease-specific measures may be more sensitive and succinct than generic measures, and hence prove convenient for general use in clinical practice. The present study sought to validate the scaling properties of the disease-specific QoL-AGHDA measure through the implementation of Rasch analysis. The study also sought to compare, by using the QoL-AGHDA, the QoL of a relatively large Swedish cohort of adults with untreated GHD with that of a reference population also from Sweden. PATIENTS: The QoL of 111 adults with untreated GHD from Stockholm and Goteborg was compared with that of 1448 adult subjects randomly selected from the population of Goteborg. MEASUREMENTS: The scaling properties of the QoL-AGHDA were assessed by investigating its fit to a dichotomous Rasch model. Rasch-transformed QoL scores from the QoL-AGHDA questionnaire were stratified by age and gender, and 95% confidence intervals were calculated. RESULTS: Rasch analysis of the QoL-AGHDA indicated the measure to be robust in terms of its unidimensionality and ordering properties, and lack of differential item functioning. The raw scores produced by the QoL-AGHDA are at the ordinal level. Non-overlapping 95% confidence intervals of Rasch-transformed interval scores in most age categories indicated that men and women with GHD had significantly lower QoL than the reference population. CONCLUSION: The Swedish QoL-AGHDA has good scaling properties, and hence can be considered a robust measure. It is suitable for assessing quality of life in adults with GH deficiency, and for making comparisons with adults who are not growth hormone deficient. Adult GH deficiency is associated with a significant impairment in QoL. PMID- 10671941 TI - Association of CTLA4 gene A-G polymorphism with type 1 diabetes in Chinese children. AB - OBJECTIVE: The CTLA4 (cytotoxic T lymphocyte associated antigen-4) gene encodes the T cell receptor involved in the control of T cell proliferation and mediates T cell apoptosis. Thus it is a strong candidate gene for T cell-mediated autoimmune disease. There is polymorphism at position 49 in exon 1 of the CTLA4 gene, providing a A-G exchange. This polymorphism is reportedly associated with type 1 diabetes in Caucasians but not in a small data set of Chinese. We wished to test this polymorphism in a larger and more homogeneous data set of Chinese children with type 1 diabetes and normal adult controls. DESIGN: A population based case-control study of a CTLA4 gene 49 A-G polymorphism was performed to look for an association with type 1 diabetes in Chinese children. PATIENTS: We analysed this polymorphism in 253 unrelated children (128 boys) with type 1 diabetes (age at diagnosis 7.1 +/- 3.7 years) and 91 randomly selected normal adults. All individuals were Han Chinese. RESULTS: The genotype and gene frequencies of children with type 1 diabetes differed significantly from those of adult controls (P = 0.0091 and P = 0.0051, respectively). Genotype CTLA4 49 G/G and G allele conferred a risk of type 1 diabetes (RR = 2.13, 95% CI = 1.31-3.46, P = 0.0022; RR = 1.68, 95% CI = 1.17-2.43, P = 0.0051, respectively). CONCLUSIONS: This study demonstrates that CTLA4 49 A-G polymorphism is associated with type 1 diabetes in Han Chinese children. The CTLA4 49 G allele confers an increased risk of type 1 diabetes. PMID- 10671942 TI - Insulin sensitivity and secretion influence the relationship between growth hormone-binding-protein and leptin. AB - BACKGROUND: A direct relationship between body mass index (BMI), visceral adipose tissue, insulin levels and growth hormone-binding protein (GHBP) activity has consistently been reported. It was recently described that GHBP directly depends on serum leptin levels. Since leptin co-varies with insulin secretion and/or sensitivity, we aimed to study the influence of these variables on plasma GHBP activity. SUBJECTS: In order to isolate the effects of obesity per se from those of insulin secretion, three groups of subjects were prospectively studied: 14 lean, 10 obese and nine obese subjects with glucose intolerance. MEASUREMENTS: The percentage of body fat was measured through bioelectric impedance. Insulin sensitivity and secretion were determined through a frequently sampled intravenous glucose tolerance test with minimal model analysis. Serum leptin was measured by radioimmunoassay. GHBP activity was determined by the high performance liquid chromatography-gel filtration method. RESULTS: Plasma GHBP activity was found to correlate with BMI (r = 0. 65, P < 0.0001), fat mass (r = 0.51, P = 0.003), waist circumference (r = 0.64, P < 0.0001), waist-to-hip ratio (r = 0.42, P = 0.01), insulin sensitivity (SI, r = - 0.61, P = 0.0001), insulin secretion (expressed as the acute insulin response to intravenous glucose, AIRg) (r = 0.48, P = 0.006) and leptin concentration (r = 0.49, P = 0.004). The associations with SI (r = - 0.42, P = 0.02) and AIRg (r = 0.38, P = 0.03) persisted even after controlling for fat mass. Since insulin secretion and insulin sensitivity usually covary in glucose tolerant subjects (an increased insulin secretion is necessary to compensate a decreased insulin sensitivity), we constructed a multiple linear regression to predict GHBP activity. In this model, SI (P = 0.005), AIRg (P = 0.02) and SD score-leptin (P = 0.03) independently contributed to 34, 10 and 8% of the variability in serum GHBP activity. CONCLUSIONS: Our results suggest that plasma GHBP activity is simultaneouslly influenced by insulin secretion and sensitivity and leptin. Perhaps leptin, through increased insulin secretion, might induce GHBP/GH secretion, explaining the normal to high insulin-like growth factor (IGF)-I levels found in overnutrition. PMID- 10671943 TI - Serum levels of growth hormone binding protein in children with normal and precocious puberty: relation to age, gender, body composition and gonadal steroids. AB - AIM: To study the regulation of GHBP serum levels by gonadal steroids in normal and precocious puberty. STUDY PROTOCOL: We studied GHBP levels in relation to age, sex, pubertal maturation, body composition as well as to circulating IGF-I and gonadal steroid levels in 320 healthy children. Furthermore, we studied the regulation of circulating GHBP in 33 girls with central precocious puberty before and during gonadal suppression with GnRH agonist. METHODS: GHBP was determined by a time-resolved fluoroimmunoassay (GHBP TR-FIA) based on a commercially available immunoassay for GH, the DELFIA GH assay. RESULTS: In healthy children GHBP levels were significantly higher in normal girls compared with boys, and there was no significant increase in GHBP in puberty in both sexes. GHBP levels did not correlate with height (SDS), age, pubertal stage, IGF-I or testosterone/oestradiol levels in boys and girls, respectively. There were significant correlations between BMI and GHBP in boys and girls (R 2 = 0.14 and R 2 = 0.12, both P < 0.0001). Furthermore, GHBP correlated highly significantly with the percentage body fat, determined by BIA in 43 healthy girls (R 2 = 0. 40, P < 0.0001). GHBP levels were significantly higher in girls with central precocious puberty (CPP) (1.31 SDS (1.26), mean (SD)) compared to prepubertal controls (P < 0.0001), and above + 2 SD in 10 out of 33 patients. In girls with CPP, GHBP correlated inversely with oestradiol before treatment (R 2 = 0.26, P < 0.01) and there was a tendency towards a positive correlation with BMI (R 2 = 0.13, P = 0.078). By contrast, there were no signficant correlations between GHBP and IGF-I or height SDS. Gonadal suppression with GnRH agonist treatment caused a transient significant increase of 0.57 SD after 2 months of treatment (P < 0.001), but decreased to baseline levels hereafter. CONCLUSION: We conclude that in children, as in adults, body fat is the primary determinant for the circulating level of GHBP, and that the difference in body fat is probably the main factor for the higher levels of serum GHBP in girls compared with boys, as well as for the negative influence of testosterone levels in boys and of oestrogen levels in girls. The elevation in GHBP levels observed in girls with central precocious puberty is probably due their higher body fat content. PMID- 10671944 TI - Administration of recombinant human growth hormone on alternate days is sufficient to increase whole body protein synthesis and lipolysis in growth hormone deficient adults. AB - OBJECTIVE: At present, the duration of the effect of recombinant human growth hormone (rhGH) on the rates of protein synthesis and lipolysis in GH deficient (GHD) adults is unknown. This study was designed to establish the frequency of rhGH administration necessary to provide the beneficial metabolic effects of the hormone in GHD adults. DESIGN AND PATIENTS: Two different studies (A and B) were performed in two groups of five GHD men. In study A, whole body protein and lipid kinetics was determined in the basal state (Bas), 12 (GH12h) and 36 (GH36h) h after the last of seven injections of rhGH (3.3 microg/kg), given at bedtime on alternate days. In study B, the same parameters were determined in the basal state (Bas), 60 (GH60h) and 84 (GH84h) h after the last of seven injections of rhGH (3.3 microg/kg), given at bedtime at 3 day intervals. MEASUREMENTS: The rates of protein metabolism were estimated by infusing [1-13C]leucine, and those of lipolysis by infusing [1,1,2,3, 3-D5]glycerol. RESULTS: Leucine oxidation decreased (P < 0.01) by approximately 30% after GH12h and GH36h but did not change after GH60h and GH84h. Non-oxidative leucine disposal increased after GH12h and GH36h by approximately 13% (P < 0.05) whereas it did not change after GH60h and GH84h. Glycerol appearance increased (P < 0. 01) by approximately 45% after GH12h and GH36h but did not change after GH60h and GH84h. CONCLUSIONS: The effects on protein and lipid metabolism following the injection of rhGH last longer than 36 and less than 60 h. In fact, rhGH administration on alternate days induced a sustained increase in the rates of protein synthesis and lipolysis of GHD adults, whereas a longer interval of administration (3 days) had no effect by 60 h. PMID- 10671945 TI - Insulin-like growth factor I administration induces fluid and sodium retention in healthy adults: possible involvement of renin and atrial natriuretic factor. AB - OBJECTIVE: Growth hormone induces fluid and sodium retention. The underlying mechanism is, however, incompletely understood. A possible mediator could be IGF I. To investigate the impact of IGF-I administration on body fluid distribution and sodium homeostasis in healthy subjects, we examined normal subjects during six days IGF-I treatment and during a six-day control period. DESIGN AND MEASUREMENTS: Eight normal male subjects aged 23-30 years were randomised to receive IGF-I 50 microg/kg subcutaneously thrice daily during a six day study period, and to a six day control period. After each study period, extracellular volume and plasma volume (ECV, PV) were determined using 82Br and 125I-albumin. Blood samples, urinary sodium excretion, and bioimpedance were measured every second day of each study period. RESULTS: Serum IGF-I (microg/l) increased during active treatment (control, 293 +/- 9; IGF-I, 628 +/- 42; P < 0.01). ECV (l) was expanded by IGF-I (control, 18.42 +/- 0.28; IGF-I, 19.72 +/- 0.50; P < 0.05) whereas PV (l) remained unaffected (control, 3.76 +/- 0.11; IGF-I, 3.80 +/- 0.16; n.s.). Likewise, bioimpedance and body weight were unchanged by IGF-I. Plasma renin (mU/l) increased but not significantly during IGF-I (control, 28.7 +/- 2.7; IGF-I, 39.9 +/- 4.3; P = 0.08), and plasma aldosterone was unaffected by IGF-I. N Terminal proANF (pmol/l) was suppressed during IGF-I administration (control, 422 +/- 32; IGF-I, 330 +/- 20; P < 0.05). Diurnal sodium excretion (mmol) was reduced during IGF-I administration (control, 151 +/- 8; IGF-I, 124 +/- 7; P < 0.05). CONCLUSION: IGF-I treatment causes fluid and sodium retention. This may be mediated by increased renin release and suppression of atrial natriuretic factor. The present data suggest that the fluid and sodium retaining effect of GH is at least partly mediated through IGF-I. PMID- 10671946 TI - Association of elevated insulin-like growth factor binding protein-1 with insulin resistance in hyperthyroidism. AB - OBJECTIVE: Insulin-like growth factor binding-protein-1 (IGFBP-1) has a role in glucose homeostasis and is present at high concentrations in hyperthyroidism. We have investigated the relationship between IGFBP-1 concentration and glucose homeostasis in hyperthyroidism. DESIGN: Patients and controls had intravenous glucose tolerance tests (IVGTT) and/or oral glucose tolerance tests (OGTT). Patients were tested when hyperthyroid and when euthyroid whilst the controls were tested once. The IVGTT was used to assess insulin sensitivity and the OGTT to establish that the study group had abnormal glucose tolerance. The hyperthyroid patients were treated with methimazole to restore euthyroidism. PATIENTS: Ten patients (9 females) and 13 healthy controls (9 females) consented to the study. Ten patients and nine controls (7 females) had IVGTT. Six patients (5 females) and six controls (4 females) had OGTT. MEASUREMENTS: Glucose, insulin, glucagon, GH and IGFBP-1 were measured during GTT. IGF-I, free thyroid hormones, and TSH concentrations were measured basally. RESULTS: Hyperthyroid subjects were insulin resistant and 67% had impaired glucose tolerance. Fasting IGFBP-1 levels were doubled in hyperthyroid subjects compared to healthy controls and correlated positively with free T4 (r = 0.84, P < 0.0001), with peak glucose during the OGTT (r = 0.68, P < 0.005) with peak insulin during the IVGTT (r = 0.51, P < 0.005) and negatively with glucose disappearance constant (r = - 0.52, P < 0.005). IGFBP-1 was highly phosphorylated in hyperthyroid and control subjects. Fasting insulin and IGFBP-1 levels were unrelated but IGFBP-1 suppressed acutely during GTT in all groups. GH levels fell less in patients with hyperthyroidism than in normals during GTTs. CONCLUSIONS: We conclude that in hyperthyroidism thyroid hormones directly increase fasting IGFBP-1 concentration but acute regulation of IGFBP-1 by insulin is normal and that elevated fasting phosphorylated IGFBP-1 concentration is associated with insulin resistance. PMID- 10671947 TI - Testicular dysfunction in men with primary hypothyroidism; reversal of hypogonadotrophic hypogonadism with replacement thyroxine. AB - OBJECTIVE: Primary hypothyroidism can cause disturbances in normal gonadal function. The aim of this study was to investigate the relationship in men between hypogonadism and primary hypothyroidism and the extent to which free and total testosterone levels rose after introduction of replacement thyroxine. DESIGN: Paired study of patients in a hypothyroid and thyroxine treated state. PATIENTS: Ten men with primary hypothyroidism. MEASUREMENTS: Free and total testosterone, gonadotrophin and prolactin levels before and after thyroxine replacement therapy. RESULTS: Low free testosterone levels (161 +/- 62 pmol/l) demonstrated at the time the men were hypothyroid rose significantly with the commencement of thyroxine replacement (315 +/- 141 pmol/l; P < 0.001). Gonadotrophin levels were not elevated consistent with hypogonadotrophic hypogonadism. Hyperprolactinaemia, which can occur in primary hypothyroidism and cause hypogonadotrophic hypogonadism, was not present in the majority of these patients. However a reduction in prolactin level was evident with thyroxine replacement and a rise in free testosterone levels. CONCLUSION: This suggests an effect of hypothyroidism on gonadotrophin secretion at the level of the hypothalamus-pituitary, either directly or through modulation of prolactin secretion. Low free testosterone may also be a contributing factor to some of the symptoms and signs of hypothyroidism in men. PMID- 10671948 TI - Parathyroidectomy for primary hyperparathyroidism induces positive uncoupling and increases bone mineral density in cancellous bones. AB - OBJECTIVE: Osteopenia is an important feature of primary hyperparathyroidism (PHP). However, little is known about the change of bone mineral density (BMD) in PHP after surgery. The aim was to investigate the mechanisms of increased BMD after parathyroidectomy in patients with PHP. DESIGN: Prospective observational study. PATIENTS: Ten patients with PHP (7 women, 3 men; mean age 53.2+/-9.1 years). All patients underwent parathyroidectomy for excision of parathyroid adenoma. MEASUREMENTS: BMDs of two cancellous bone-rich sites (L2-L4 lumbar spine and ultra-distal end of the radius, RUD) and one cortical bone-rich site (distal third of the radius, R33%) were measured using dual energy X-ray absorptiometry, before, and 3, 6 and 12 months after surgery. Serum intact PTH, intact osteocalcin, bone type alkaline phosphatase (b-ALP), alkaline phosphatase, calcium, and urinary deoxypyridinoline (Dpd) were measured before, and 1 and 3 days, and 1, 2, 3, 4, 6, 8, 12, and 24 weeks after surgery. RESULTS: Parathyroidectomy resulted in a significant increase in BMDs of L2-L4 and RUD at 3 months postoperatively. Urinary Dpd levels decreased within a few days after surgery, while b-ALP and osteocalcin decreased more slowly throughout the first few months after surgery. The ratio of osteocalcin/Dpd at 1 week after surgery correlated significantly with the percentage change in BMD of L2-L4 at 3 and 6 months after surgery. The ratio of osteocalcin/Dpd at 2 weeks correlated significantly with the percentage change in BMD of L2-L4 at 3, 6 and 12 months after surgery. The preoperative values of osteocalcin, b-ALP, PTH and calcium were positively correlated with the change in BMD of RUD at 3 months and L2-L4 at 12 months, RUD at 6 months, RUD at 3 months and L2-L4 at 12 months, respectively. CONCLUSIONS: In primary hyperparathyroidism patients, the major increase in bone mineral density following parathyroidectomy occurs within 3 months. Parathyroidectomy resulted in a marked increase in bone mineral density of cancellous bones compared to that of cortical bones. The early increase in bone mineral density was due to a preferential activation of bone formation over bone resorption as evidenced by changes in bone metabolic markers. Our results also showed that the preoperative levels of bone metabolic markers may predict the gain in bone mineral density after parathyroidectomy. PMID- 10671949 TI - Consequences of vitamin D receptor gene polymorphisms for growth inhibition of cultured human peripheral blood mononuclear cells by 1, 25-dihydroxyvitamin D3. AB - OBJECTIVE: In the vitamin D receptor (VDR) gene a BsmI restriction fragment length polymorphism (RFLP) in intron 8 and a translational start-site polymorphism, identified as a FokI RFLP, have been described. Crucial for a proper interpretation of these polymorphisms in association studies is the knowledge whether they have direct consequences for 1,25-(OH)2D3 action at cellular level. The present study was designed to assess functional significance of the FokI and BsmI VDR gene polymorphisms in peripheral blood mononuclear cells (PBMC) with a natural occurring VDR genotype for cell growth inhibition by 1,25 (OH)2D3. DESIGN: PBMC of women were isolated, VDR genotyped and in vitro inhibition by 1,25-(OH)2D3 of Phytohemagglutinin (PHA)-stimulated growth of PBMC was examined in relation to VDR genotype. RESULTS: PHA-stimulated growth and maximal growth inhibition were independent of VDR genotype. However, the FF genotype had a significant lower ED50 than the Ff genotype corresponding to an allele dose effect of 0.32 nM per f allele copy (P = 0.0036). For BsmI genotypes no differences in ED50 were observed. CONCLUSION: The present study demonstrates for the first time in cells with a natural VDR genotype a direct functional consequence of the VDR gene translational start-site polymorphism for the action of 1,25-(OH)2D3. Especially under conditions of vitamin D insufficiency these findings might have clinical implications. PMID- 10671950 TI - Nasogastric feeding enhances nutritional status in paediatric liver disease but does not alter circulating levels of IGF-I and IGF binding proteins. AB - OBJECTIVE: Complications of childhood cirrhosis include abnormal growth and malnutrition, associated with abnormalities in circulating IGFs and IGFBPs. Controlled studies suggest that intensive enteral feeding enhances nutritional status. The aim was to ascertain whether nasogastric feeding improves nutritional status in clinical practice and to assess the effect of feeding on serum IGF-I and IGFBPs. PATIENTS: Thirty-three children (median age 0.6 years) with biliary atresia and failure to thrive who were treated with nasogastric feeding. MEASUREMENTS: Height, weight and triceps skin fold thickness were measured prior to feeding and regularly for 1 year or until feeding was stopped. Serum IGF-I and IGFBPs were measured by immunoassay at the same intervals. RESULTS: The median duration of feeding was 3.7 months. Twenty-two stopped feeding after liver transplantation, while 10 stopped electively and 1 boy died. Before feeding, the children were losing weight and height centile. Triceps skin fold thickness, weight and height SD scores improved with feeding. Baseline serum IGF-I and IGFBP 3 were low, while IGFBP-1 and IGFBP-2 were raised. IGF-I and IGFBP-1 did not change with feeding. IGFBP-2 fell and reached a nadir by 3 months, while IGFBP-3 rose temporarily for 4-6 weeks. CONCLUSIONS: Nasogastric feeding improves body composition in paediatric liver disease but circulating IGF-I and IGFBPs remain abnormal and do not play a major role in mediating these changes. This does not exclude a paracrine or autocrine effect of IGF-I. PMID- 10671951 TI - Raloxifene reduces atherosclerosis: studies of optimized raloxifene doses in ovariectomized, cholesterol-fed rabbits. AB - OBJECTIVE: We have previously shown that raloxifene, a selective oestrogen receptor modulator, 35 mg/day inhibits atherosclerosis in ovariectomized, cholesterol-fed rabbits. This effect was only partial as compared to 17beta oestradiol 4 mg/day; however, plasma raloxifene concentrations were low relative to those obtained in raloxifene-treated women. We therefore investigate the effects of raloxifene at higher doses. DESIGN: The study on atherosclerosis in ovariectomized, cholesterol-fed rabbits (n = 80) compared raloxifene 70 mg/day and 210 mg/day to 17beta-oestradiol 4 mg/day and placebo. RESULTS: After 48 weeks of therapy, the aortic cholesterol content in the 70 mg/day and 210 mg/day raloxifene treatment groups were 471 +/- 56 nmol/mg protein and 456 +/- 56 nmol/mg protein, respectively. This was significantly less than in the placebo group (654 +/- 69 nmol/mg protein; P < 0.05). In the oestrogen-treated group, the aortic cholesterol content was 357 +/- 62 nmol/mg protein (P < 0.01 as compared to placebo). Differences in serum lipids between the treatment groups could only partly explain the effect on aortic cholesterol content, indicating that additional anti-atherogenic mechanisms may contribute to the decrease in aortic atherosclerosis. This anti-atherosclerotic activity of raloxifene was observed at plasma concentrations comparable to those in postmenopausal women during raloxifene treatment. CONCLUSIONS: We conclude that clinically relevant raloxifene treatment inhibits aortic atherosclerosis in ovariectomized, cholesterol-fed rabbits. PMID- 10671952 TI - GIP-dependent adrenal Cushing's syndrome with incomplete suppression of ACTH. AB - ACTH-independent Cushing's syndrome may be due to the development of ectopic hormone receptors in adrenal tissue. Thus, in food-dependent Cushing's syndrome the adrenals aberrantly express receptors for gastric inhibitory polypeptide (GIP). We present the case of a 60-year-old woman with food-dependent Cushing's syndrome whose cortisol levels increased after stimulation with CRH. In this patient with Cushing's syndrome the finding of low basal plasma cortisol levels in the late night and early morning as well as a paradoxical rise of plasma cortisol during a 7-h infusion with dexamethasone (carried out without any restriction in food intake), suggested that cortisol production was stimulated at times of food intake. Hourly measurements of plasma cortisol for 48 h revealed prominent meal-related peaks. A plasma cortisol response, elicited by oral glucose administration, could be prevented by octreotide. Plasma ACTH was low or undetectable. CRH administration was followed by a ACTH response from 3 to 16 ng/l and a plasma cortisol response from 230 to 680 nmol/l. Octreotide treatment for nearly five months induced a partial clinical and biochemical remission. Total bilateral adrenalectomy was performed. The left adrenal was grossly enlarged (7 x 5.5 x 4 cm) and the right adrenal was slightly enlarged (6 x 4 x 1.8 cm). Microscopy revealed bilateral nodular hyperplasia. Cell suspensions of adrenal tissue from the patient did respond in a dose-dependent fashion to stimulation with GIP and were very sensitive to stimulation with synthetic ACTH1 24. However, CRH had no significant effect on cortisol production in vitro. Using RT-PCR amplification and cDNA hybridization, GIP receptor was found to be overexpressed in the left and right adrenal tissues from this patient as compared to adrenal tissues from a normal individual or from non GIP-dependent adrenal Cushing's syndrome. There was no evidence of presence of adrenal CRH receptors. Thus, in this patient with food-dependent Cushing's syndrome, the CRH-induced plasma ACTH and cortisol response is probably mediated by an incomplete suppression of the HPA axis as a result of the intermittent food-dependent nature of Cushing's syndrome. PMID- 10671953 TI - Complete resolution of protease inhibitor induced diabetes mellitus. PMID- 10671954 TI - Metformin and polycystic ovary syndrome. PMID- 10671955 TI - Metformin and ovarian steroidogenesis in PCOS women. PMID- 10671956 TI - Metformin and polycystic ovary syndrome PMID- 10671957 TI - Carbenoxolone effects in congenital adrenal hyperplasia. PMID- 10671958 TI - Carbenoxolone effects in congenital adrenal hyperplasia PMID- 10671959 TI - Screening for diabetes in obese women: comparison between the New American Diabetes Association and WHO criteria. PMID- 10671960 TI - Interferon-alpha therapy for melanoma. AB - Although surgery may be curative in early malignant melanoma, its effect on survival lessens with each succeeding stage of the disease. A wide variety of immunological strategies have therefore been used to improve the prognosis of patients with malignant melanoma, but adjuvant therapy with interferon (IFN) alpha is the only treatment to show a therapeutic benefit in randomized controlled studies. The current data indicates that where IFN-alpha is used at low dose, its main effect is on disease-free survival, whereas high-dose regimens may improve overall survival as well. This paper will review the published data on IFN-alpha therapy in patients with intermediate and high-risk melanoma and explore future avenues for managing patients with this difficult disease. PMID- 10671961 TI - Treatment options for giant congenital naevi. AB - Giant congenital naevi (GCN) are disfiguring, potentially malignant pigmented naevi present at birth. The naevus cells in GCN are found throughout the dermis and sometimes penetrate the subcutaneous septa. It is claimed that superficial, more heavily pigmented and biologically different naevus cells reside in the upper dermis. Partial removal of these superficial naevus cells by dermabrasion, laser therapy, curettage or shave excision is less traumatic than excision surgery and produces an acceptable cosmetic result. However, none of these techniques or excision of GCN to superficial fat completely removes the risk of malignant transformation. PMID- 10671962 TI - An evaluation of educational requirements for community nurses treating dermatological patients. AB - In a questionnaire to community nurses treating dermatological patients, 14 out of 69 (20%) either treated children or gave advice to parents regarding childhood eczema, 35 (51%) treated adult eczema, 11 (16%) treated psoriasis, 55 (80%) treated leg ulcers, and 30 (43%) treated other dermatological problems. Specific questions regarding confidence to treat or educate were analysed in relation to the tasks being performed. All but 15% (8/55) treating leg ulcers were confident about their ability to apply four-layer bandaging. However, 8 out of 11 (72%) respondents treating psoriasis were not confident about their ability to treat scalp scaling, 11 out of 14 (79%) of those treating childhood eczema were not confident about applying body suiting, and 26 out of 36 (72%) of those treating eczema (any age), were not confident about ability to recognize infection as a cause or complication of dermatoses. The favoured educational modalities were visits to the local dermatology department (60/69, 87%), availability of a dermatology Nurse Practitioner or Liaison Nurse, or access to a hospital nurse run dermatology clinic (both 44/69, 63%), or attendance at courses (36/69, 52%). Community nurses have an important role in treating and educating patients who may not require or be able to attend hospitals for treatment; they will achieve this best by provision of relevant locally based education, with allocation of adequate study time. PMID- 10671963 TI - Chloroxylenol and zinc oxide containing cream (Nels cream) vs. 5% benzoyl peroxide cream in the treatment of acne vulgaris. A double-blind, randomized, controlled trial. AB - Forty-one subjects completed a double-blind controlled randomized study comparing the following: (i) Nels cream (containing chloroxylenol and zinc oxide); (ii) 5% benzoyl peroxide cream; and (iii) the vehicle of the Nels cream. Patients applied the medications twice daily for 8 weeks. At the end of the test period there was no significant difference in the reduction of inflammatory and noninflammatory lesion counts achieved by Nels cream and benzoyl peroxide. Both creams proved superior to the vehicle. Efficacy grading by subjects and investigators showed no significant difference between Nels cream and benzoyl peroxide. However, side effects such as peeling and dryness caused by the treatment were significantly less in the Nels cream group. PMID- 10671964 TI - Yellow nail syndrome: the nail that grows half as fast grows twice as thick. AB - We report a case of a 51-year-old man with yellow nail syndrome (YNS).1 During a 23-week period of study, the dynamics of thumbnail growth were compared between one affected thumb and the normal contralateral thumb. Longitudinal nail growth was normal (0.46 mm/week) in the normal thumb and double that of the affected thumb (0.23 mm/week). Thickness of nail at the free edge in the affected thumb (0.97 mm) was twice that of the normal thumb (0.57 mm). Within the nail plate in the dorso-ventral axis there were 50% more cells in the affected thumb (358) in comparison with the contralateral control (242). This case illustrates that rate of longitudinal growth does not necessarily reflect nail plate production. PMID- 10671965 TI - Pulmonary toxicity in a patient with psoriasis receiving methotrexate therapy. AB - We report a 34-year-old woman with psoriasis who developed shortness of breath during methotrexate therapy. Methotrexate had been started 4 months earlier and the patient had ingested a cumulative dose of 232 mg. Pulmonary function tests showed a reduction in transfer factor to 76% of predicted. Methotrexate was stopped and her symptoms rapidly resolved. Pulmonary function tests deteriorated further despite stopping methotrexate but with no recurrence of symptoms with a transfer factor of 66% of predicted 2 months later. At 5 months after stopping methotrexate the patient remained well and pulmonary function had improved with a transfer factor of 79% of predicted. Pulmonary toxicity is a rare but important adverse effect of methotrexate therapy in patients with psoriasis. PMID- 10671966 TI - Welding and non-melanoma skin cancer. AB - This paper reports on five cases of nonmelanoma skin cancer (NMSC) occurring in welders, an occupation in which there is potential for intense exposure to nonsolar ultraviolet irradiation. The status of ultraviolet light as the major aetiological factor in NMSC is well established, and in the great majority of cases solar irradiation is the source. The possible role of nonsolar sources of ultraviolet irradiation should not be overlooked. PMID- 10671967 TI - Protean manifestations of lipoid proteinosis in a 16-year-old boy. AB - We report a 16-year-old Japanese male with lipoid proteinosis showing various skin manifestations. The patient was born to nonconsanguineous parents and none of his relatives was similarly affected. The patient suffered from a hoarse voice and refractory temporal epilepsy from early childhood. Computed tomography scanning of the brain showed bilateral calcification in the temporal lobes, a characteristic feature of lipoid proteinosis. On physical examination, various skin manifestations, including papules and haemorrhagic blisters, acne-like scars at sites of minor trauma or friction, and beads of small papules along the free margins of the eyelids were noted. A skin biopsy showed deposits of homogeneous hyaline-like material, positive on periodic acid-Schiff staining, throughout the dermis, particularly around small blood vessels. It is noteworthy that a range of characteristic skin lesions can be present in a patient with lipoid proteinosis even with mild systemic involvement. PMID- 10671968 TI - Renal angiogram abnormalities in a case of cutaneous polyarteritis nodosa. AB - The existence of a limited cutaneous form of polyarteritis nodosa remains controversial. It has been characterized and contrasted with systemic polyarteritis nodosa by running a chronic course and having a relatively good long-term prognosis. We report a case with clinical features fitting the criteria for cutaneous polyarteritis nodosa but also showing evidence of associated renal aneurysms on selective visceral angiography. These findings suggest that the differentiation between systemic and cutaneous forms might be an oversimplification and we discuss the relationship between the two forms. PMID- 10671969 TI - Oesophageal lichen planus. AB - We present a case of lichen planus affecting the oesophagus of an 80-year-old woman. Symptomatically, the lesions manifested themselves as dysphagia requiring repeated oesophageal dilatations. The patient was unable to tolerate several conventional treatments and so pulsed methylprednisolone was given with some beneficial short-term effects. Due to potential for malignant change in lichen planus of the mucous membranes it is important to remember this complication and investigate patients with oesophageal symptoms. PMID- 10671970 TI - Combined cutaneous hamartoma encompassing benign melanocytic naevus, vellus hair cyst and epidermoid cyst. AB - We describe a combined cutaneous hamartoma in a 57-year-old man, which comprised a vellus hair cyst, an epidermoid cyst and an intradermal melanocytic naevus in a single facial tumour. The vellus hair cyst was filled with keratinous material with unusual slit-like lacunae. While numerous cases of epidermoid cyst have been reported in association with an intradermal melanocytic naevus, our case is a rare example of a lesion combining several hamartomatous elements. PMID- 10671971 TI - Epidermodysplasia verruciformis: association with isolated IgM deficiency and response to treatment with acitretin. AB - We describe a 25-year-old woman, who had extensive, large viral warts consistent with epidermodysplasia verruciformis (EV) since she was 6-year-old. Laboratory studies revealed an isolated IgM-deficiency, but the patient demonstrated no other abnormalities. She was treated with oral acitretin (0.5-1 mg/kg/day) for six months and her skin lesions improved slightly. However, after discontinuing the treatment, the lesions came back but she declined further treatment. PMID- 10671972 TI - Association of multiple familial cutaneous leiomyoma with a uterine symplastic leiomyoma. AB - We describe a patient who has familial cutaneous leiomyoma in association with a symplastic uterine leiomyoma. This association has not been described previously. PMID- 10671973 TI - Facial cutaneous tuberculosis: an unusual presentation. AB - Periocular cutaneous tuberculosis is a rare occurrence. We describe a 75-year-old Caucasian woman with tuberculosis of the left periocular region which responded completely to standard antituberculosis therapy. We hypothesize this unusual presentation may be due to minor trauma followed by inoculation. PMID- 10671974 TI - Phakomatosis pigmentovascularis IIb with renal anomaly. AB - Phakomatosis pigmentovascularis (PPV) is a rare congenital naevoid syndrome; most case reports originate in Japan. The major clinical manifestations consist of coexisting extensive naevus flammeus and pigmentary naevus with or without systemic involvement. We report an 8-year-old Taiwanese boy, who was born with extensive naevus flammeus and other anomalies comprising persistent aberrant Mongolian spot-like pigmentary patches, leg-length discrepancy, pelvic obliquity, scoliosis and bilateral melanosis oculi bulbi. Further investigation also revealed agenesis of the right kidney. The cutaneous lesions remained unchanged over a 3-year follow-up period. Within the classification of PPV, this boy's disorder represents an example of PPV IIb. Right kidney agenesis, which has never been observed in PPV, may be an incidental finding. PMID- 10671975 TI - Subcutaneous sarcoidosis associated with vitiligo, pernicious anaemia and autoimmune thyroiditis. AB - We report a patient with pernicious anaemia, primary autoimmune hypothyroidism and vitiligo, who presented with subcutaneous nodules. Histopathology of the nodules revealed noncaseating granulomas, consistent with a diagnosis of sarcoidosis. Mild pulmonary sarcoid was also detected. Although an association between sarcoidosis and other autoimmune diseases is well-recognized, the presence of the particular autoimmune diseases in our patient and the involvement of subcutaneous fat in the sarcoidal inflammation, appears to represent a most unusual clinicopathological combination. PMID- 10671976 TI - Bacterial superantigens and inflammatory skin diseases. AB - Bacteria seem to play an important role in the induction and maintenance of inflammatory skin diseases such as psoriasis and atopic dermatitis. Toxins from bacteria including Streptococcus and Staphylococcus aureus, have been shown to function as a new type of allergen termed 'superantigen'. Superantigens bypass the normal control of T-cell activation and activate all T-cell clones bearing certain types of variable chain on the T-cell receptor: this leads to vigorous T cell activation and cytokine release. These bacterial superantigens may be involved in induction and aggravation of inflammatory skin diseases. Guttate psoriasis is often preceded by a streptococcal throat infection and T cells specific for streptococcal superantigens have been identified in the skin of patients. The skin of patients with atopic dermatitis is often colonized with superantigen-releasing Staph. aureus, and application of a staphylococcal superantigen to human skin induces an eczematoid reaction. PMID- 10671977 TI - Population-based surveys on the frequency of common skin diseases in adults--is there a risk of response bias? AB - Population-based surveys on the frequency of common skin diseases are important in determining the health needs of a community. As they rely on voluntary presentation, there is a risk of response bias which may compromise the quality of the data obtained. The aim was to determine in what way and to what degree response bias may occur in a population-based survey on the frequency of common skin diseases in adults. A follow-up study was conducted on 1043 out of 2500 adults who did not attend for examination as part of a randomized population based survey on the frequency of common skin diseases amongst adults in Maryborough, Central Victoria, Australia. Nonrespondents were more likely to be at the extremes of age, retired, unemployed and less likely to report that they had a history of skin disease than the respondents. Subsequent examination of a sample of the nonrespondents revealed they were more likely to have skin cancers and Campbell de Morgan angiomas than the respondents. These differences cannot be fully explained on the basis of an age-related response bias alone. Response bias is a risk in population-based surveys of common skin diseases which rely on voluntary presentation. Some attempt should be made to sample the nonrespondents in these surveys to determine the nature and extent of any bias and to adjust for it, if necessary. PMID- 10671978 TI - Evaluation of a contact allergy clinic. AB - This study aims to assess the value of patch testing in a specialist contact allergy clinic as compared to testing carried out by general dermatologist for the diagnosis of contact allergy. One hundred consecutive patients referred for patch testing were tested to the European standard and specialized series. A comparison was made between patch tests read and interpreted by a consultant dermatologist with an interest in contact allergic dermatitis and by an experienced registrar who had not yet received any formalized specialized training. Results of testing with additional allergens acquired following the establishment of the specialist clinic were examined. In addition patients with a hand problem and/or symptoms suggestive of contact urticaria underwent skin prick testing. There was a variation in patch test reading and interpretation between the consultant and registrar in 10 patients. Patch testing using allergens from the additional series revealed additional relevant contact allergies in 19 patients. In 14 patients, allergies would have been missed by a limited additional series. Contact urticaria was detected in seven patients. This study has confirmed the value of a specialist contact clinic in the diagnosis of contact allergy. The importance of formalized training in patch test reading and interpretation, testing with additional series and prick testing for the diagnosis of contact urticaria are highlighted. PMID- 10671979 TI - Two type XVII collagen (BP180) mRNA transcripts in human keratinocytes: a long and a short form. AB - We have analysed BP180 mRNA expression in normal human keratinocytes. Here we report the presence in normal keratinocytes of two COL17A1 transcripts which differ by 0.6 kb in length. Both mRNAs hybridized on Northern blot with probes directed to sequences encoding intracellular and extracellular fragments of BP180. By BLAST homology search alignments we extended the 3' untranslated region (3'UTR) of the known BP180 mRNA sequence by 877 bases to completion. Three of 20 cDNAs identified by BLAST searches contained a 610 bp deletion in this new 3'UTR sequence. Northern blot analysis with a probe complementary to this deleted sequence showed binding only to the larger mRNA. The deletion of 610 nucleotides in the smaller mRNA was verified by reverse transcription-PCR and sequencing. Genomic PCR showed the new sequence to be an extension of exon 56 of the COL17A1 gene which suggests that the second mRNA is generated by differential splicing. In normal keratinocytes the level of the smaller transcript was 5-15% of that of the larger transcript whereas in a squamous cell carcinoma cell line this ratio was reversed, the smaller mRNA being three times more abundant than the larger mRNA. The biological significance of this newly identified transcript in protein synthesis and tissue expression or in cell differentiation, proliferation or adhesion is as yet unknown. PMID- 10671980 TI - A 20-year epidemiological study of cutaneous melanoma in the Rijeka district of Croatia. AB - A retrospective study was made of the incidence of cutaneous melanoma among the population of the district of Rijeka (Croatia) during the period 1977-96. A total of 397 patients with cutaneous melanoma was documented during this period. Over the 20-year period the incidence of the tumour increased, the mean annual rate being 4. 8 (4.0 in males and 5.4 in females) in the first 10-year period and 7.16 in the second (7.1 in males and 7.3 in females). The number of registered cases in males and females was almost identical (1 : 1.05). The number of melanoma cases increased with age in both sexes, whereas it was rare in children. The most affected anatomical location was the trunk in males and the lower limbs in females. The results indicate the need for active prevention and educational programmes in this population. PMID- 10671981 TI - Choosing a dermatological hero for the millennium. Erasmus Wilson (1809-1884). PMID- 10671982 TI - Erasmus Wilson (1809-1884). PMID- 10671983 TI - Choosing a dermatological hero for the millennium. Robert Willan (1757-1812). PMID- 10671984 TI - Choosing a dermatological hero for the millennium. Hippocrates of Cos (460-377 BC). PMID- 10671985 TI - Choosing a dermatological hero for the millennium. Daniel Turner (1667-1741). PMID- 10671986 TI - Choosing a dermatological hero for the millennium. Jean-Louis Alibert (1768 1837). PMID- 10671987 TI - Choosing a dermatological hero for the millennium. William Shakespeare (1564 1616). PMID- 10671988 TI - Autologous blood transfusion. PMID- 10671989 TI - Lifetime treatment costs of beta-thalassaemia major. AB - Beta-thalassaemia major is a serious genetic disorder, which results in a considerable increase in both acute and chronic morbidity, and mortality. Although beta-thalassaemia major is a rare disease affecting approximately 600 people in the UK, treatment is intensive and predictions of the costs incurred may aid health care planning. In this report, the cost to the health service of providing treatment services for beta-thalassaemia major patients, over the course of a lifetime, is calculated in order to assist resource allocation decisions. A cost model was developed, incorporating data from disparate sources. The undiscounted lifetime cost of treating a beta-thalassaemia major patient was estimated to be pound 803,002, although when the costs were discounted at a rate of 6%, the lifetime cost was reduced to pound 219,068. Within sensitivity analyses, the discounted cost ranged from approximately pound 188,000 to pound 226,000. This report may act as a guide to those involved in the planning of health care provision with regard to the resources required to treat beta thalassaemia major patients. Such information may also be incorporated into the decision-making process for the provision of antenatal screening programmes for beta-thalassaemia major. PMID- 10671990 TI - Highly fluorescent reticulocyte count predicts haemopoietic recovery after immunosuppression for severe aplastic anaemia. AB - Highly fluorescent reticulocyte (HFR) counts are the most reliable and sensitive index of haemopoietic recovery after bone marrow or peripheral blood stem cell transplantation. We report the behaviour of HFRs during haemopoietic recovery in two patients who were affected by severe aplastic anaemia (SAA) and treated with horse antithymocyte globulin (ATG), cyclosporin A (CsA) and granulocyte colony stimulating factor (G-CSF). A HFR value > 5% of the total reticulocyte count, a reticulocyte count > 30 x 10(9)/l, and a polymorphonuclear (PMN) count > 0.5 x 10(9)/l were found after 9 and 8, 20 and 46, and 16 and 22 days, respectively, after the end of ATG. HFR recovery to > 5% anticipated the rise of PMN > 0.5 x 10(9)/l by at least 7 and 14 days, respectively. Thus, HFR evaluation could be used as a reliable and early marker of response to immunosuppression in severe aplastic anaemia. PMID- 10671991 TI - Microvolume fluorimetry for the determination of absolute CD4 and CD8 lymphocyte counts in patients with HIV: a comparative evaluation. AB - This study compared CD4 and CD8 lymphocyte counts obtained by microvolume fluorimetry (MVF) with those derived by flow cytometry (FC). Samples from 192 patients with known or suspected HIV were analysed, and the distribution of CD4 counts for these samples ranged from 0 and 1,279/microl, with 142/192 (74%) of the samples having CD4 values of less than 400/microl. Good agreement between FC and MVF CD4 counts was found (MVF = 0.98 x FC + 7.30) although there was a minor constant inter-method bias of approximately +7 cells/microl for the MVF data. For CD8 counts there was a constant bias between the two methods of approximately +23 cells/microl for FC. Most outliers were associated with higher FC CD8 counts. Supplementary analyses showed a high level of agreement between FC and MVF methods for the CD4:CD8 ratios (MVF = 0.98 x FC). This suggests that observed discrepancies between FC and MVF methods were almost certainly a result of the influence of the absolute lymphocyte counts obtained from the haematology analyser. The results confirm that the IMAGN 2000 microvolume fluorimeter system can be used as an alternative to conventional flow cytometry for the enumeration of CD4 and CD8 counts. PMID- 10671992 TI - Wiskott Aldrich syndrome presenting as congenital thrombocytopenia. AB - The application of molecular biology to haematology has provided the opportunity to revisit previous diagnoses, many of which can now be redefined. This report is on a local family previously diagnosed and published as having X-linked thrombocytopenia in 1974, and shows how the application of molecular screening has confirmed the true diagnosis of Wiskott Aldrich syndrome. PMID- 10671993 TI - Factor V Leiden and allogeneic bone marrow transplantation: chimerism as a confounding factor in genetic test interpretation. AB - Factor V Leiden is one of the most common genetic conditions predisposing to venous thrombosis. Diagnosis is currently made by plasma activity assay for activated protein C (APC) resistance or polymerase chain reaction (PCR)-based DNA assay. The occurrence of factor V Leiden is reported in a patient affected by acute myeloid leukaemia submitted to allogeneic bone marrow transplantation from an HLA identical sister. The donor was not affected by the factor V mutation. The patient did not develop thrombosis during induction and consolidation chemotherapy and the post-transplantation course was not complicated by thrombosis or veno-occlusive disease. At engraftment, PCR analysis showed the disappearance of factor V Leiden. Genetic tests on DNA after allogeneic marrow transplantation should be carefully interpreted as a result of donor chimerism. PMID- 10671994 TI - Paediatric myelodysplastic syndrome (MDS) and juvenile chronic myelogenous leukaemia (JCML) detected by cytogenetic and FISH techniques. AB - This report presents two rare cases, one of paediatric myelodysplastic syndrome (MDS) and the other juvenile chronic myeloid leukaemia (jCML). In the first case, there were clinical and biological features of MDS-refractory anaemia with excess blasts (RAEB). The bone marrow (BM) karyotype demonstrated a monosomy 7 which was confirmed by fluorescence in situ hybridization (FISH). In addition, FISH analysis showed that an alpha-satellite DNA sequence had been transferred from chromosomes 13/21 to one homologue of chromosomes 22. The BCR-ABL rearrangement was negative. In the second case, at diagnosis, the karyotype was 46,XX. FISH analysis with the simultaneous and individual application of abl and bcr probes for chromosome 9 and 22, respectively, revealed the presence of the BCR-ABL rearrangement in addition to an extra ABL sequence locating chromosome 20. A clone that was BCR-ABL gene rearrangement negative but with an extra ABL DNA sequence on chromsome 20, and another clone that was BCR-ABL gene rearrangement negative were detected by DC-FISH and uni-colour (UC-) FISH analysis. No monosomy 7 was detected by conventional cytogenetic or FISH analyses. PMID- 10671995 TI - Delayed haemolytic transfusion reaction caused by anti-M antibody in a patient receiving interleukin-2 and interferon for metastatic renal cell cancer. AB - Anti-M is usually a naturally occurring cold-reactive immunoglobulin M (IgM) antibody, often with an immunoglobulin G (IgG) component, and is seldom implicated in delayed haemolytic transfusion reactions (DHTR). However, cases have been reported. In the majority, a DHTR is not suspected until further blood is requested and a new antibody is detected on pretransfusion testing. We describe the case of a young man receiving therapy with interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) for metastatic renal cell cancer who developed a clinically suspected DHTR that was confirmed serologically to be caused by anti M, reactive at 37 degrees C. We discuss the possible role of his biochemotherapy in the development of the DHTR. PMID- 10671996 TI - Sickle cell disease and nitrous oxide-induced neuropathy. AB - We report three cases of peripheral neuropathy in patients with sickle cell disease. All had a history of frequent painful crises and regular attendance at our Accident and Emergency department where nitrous oxide analgesia was administered for prolonged periods. All three patients (one male and two females) presented with difficulty in walking associated with paraesthesiae, and neurological examination revealed signs compatible with a peripheral sensorimotor neuropathy, later confirmed by nerve conduction studies. Serum vitamin B12 levels were mildly reduced in two patients and very low in one patient (< 10 ng/l). Haemoglobin levels in all the patients were unchanged compared with their steady state levels but one had developed a macrocytosis (103 fl). Schilling tests were normal in two patients, and two patients had negative gastric parietal antibodies. All three patients were given intramuscular vitamin B12 in addition to avoiding further exposure to nitrous oxide, and their neurological symptoms improved completely. As nitrous oxide is known to cause a neuropathy similar to that seen in pernicious anaemia, we postulate that nitrous oxide analgesia combined with low B12 levels was the cause of the marked neuropathy in these patients. As a result of our observations and the probable association, we now do not use nitrous oxide analgesia in the management of patients with sickle cell disease. PMID- 10671997 TI - Systemic lupus erythematosus presenting with haemorrhagic manifestation. AB - A 26-year-old female presented with an episode of severe mucus membrane bleeding. Investigations revealed prolonged prothrombin time (PT), and partial thromboplastin time (PTT), normal thrombin time (TT) and reptilase time, thrombocytopenia, a positive test for lupus anticoagulant (LA), as well as anti cardiolipin antibodies (ACL). A toxicology screen for toxic drugs and coumadin was negative. Coagulation factor assays revealed low levels for factor II and XII. Low level inhibitor to factor II was demonstrated. Patient had a negative VDRL test and positive anti-nuclear antibodies (ANA). The diagnosis of acquired hypoprothrombinaemia secondary to circulating inhibitor induced by LA was made, and then the patient was started on prednisone, which led to cessation of the bleeding and normalization of PT and PTT, as well as an increase of factor II and factor XII levels. A few months later, the patient developed arthralgia and alopecia, and antibodies against double-stranded DNA were detected, and the diagnosis of systemic lupus erythematosis (SLE) was confirmed. The patient continued to have mild prolongation of PT and PTT while on a low dose of prednisone, but she had no bleeding symptoms. A computed tomography scan of the brain was carried out for unexplained central nervous system (CNS) symptoms, and it revealed mild hydrocephalus, which was thought to be part of the CNS manifestations of SLE. It was concluded that patients with SLE may present with haemostatic defects that are a result of either platelet-related causes (quantitative or qualitative) or coagulation factor deficiency secondary to circulating inhibitor, or both, in the absence of other features of SLE which may appear later. PMID- 10671998 TI - A change in reaction specificity of sheep liver serine hydroxymethyltransferase. Induction of NADH oxidation upon mutation of His230 to Tyr. AB - Both serine hydroxymethyltransferase and aspartate aminotransferase belong to the alpha-class of pyridoxal-5'-phosphate (pyridoxalP)-dependent enzymes but exhibit different reaction and substrate specificities. A comparison of the X-ray structure of these two enzymes reveals that their active sites are nearly superimposable. In an attempt to change the reaction specificity of serine hydroxymethyltransferase to a transaminase, His 230 was mutated to Tyr which is the equivalent residue in aspartate aminotransferase. Surprisingly, the H230Y mutant was found to catalyze oxidation of NADH in an enzyme concentration dependent manner instead of utilizing L-aspartate as a substrate. The NADH oxidation could be linked to oxygen consumption or reduction of nitrobluetetrazolium. The reaction was inhibited by radical scavengers like superoxide dismutase and D-mannitol. The Km and kcat values for the reaction of the enzyme with NADH were 74 microM and 5. 2 x 10-3 s-1, respectively. This oxidation was not observed with either the wild type serine hydroxymethyltransferase or H230A, H230F or H230N mutants. Thus, mutation of H230 of sheep liver serine hydroxymethyltransferase to Tyr leads to induction of an NADH oxidation activity implying that tyrosyl radicals may be mediating the reaction. PMID- 10671999 TI - A switch in the direction of the effect of insulin on the partitioning of hepatic fatty acids for the formation of secreted triacylglycerol occurs in vivo, as predicted from studies with perfused livers. AB - The direct effects of insulin on hepatic triacylglycerol secretion are important because they may determine the degree of postprandial hyperlipidaemia, a known risk factor for the development of atherosclerotic lesions. Previous work from this laboratory, conducted on isolated perfused rat livers [Zammit, V.A., Lankester, D.J., Brown, A.M. & Park, B.S. (1999) Eur. J. Biochem. 263, 859-864], has indicated that the effect of insulin on hepatic triacylglycerol secretion is dependent on the prior physiological state of the donor animals. In this paper, we demonstrate that a switch in the direction of insulin action on hepatic partitioning of fatty acyl moieties towards triacylglycerol secretion also occurs in vivo between the fed, normoinsulinaemic state and the fasted or severely insulin-deficient states. The partitioning of fatty acids in the liver of awake, unstressed rats was studied using selective labelling of hepatic fatty acids during hyperinsulinaemic-euglycaemic clamps achieved through the use of hepatocyte-targeted liposome-encapsulated insulin preparations. The data show that, whereas in the fed, normoinsulinaemic state, insulinization of the liver raises the proportion of fatty acids directed towards secreted triacylglycerol, in the fasted or insulin-deficient states, insulin inhibits the partitioning of acyl moieties into secreted triacylglycerol. These data show that observations on the direction of insulin action on hepatic triacylglycerol secretion obtained using isolated perfused rat livers are reflected in the effects of the hormone on hepatic fatty acid partitioning in vivo. They offer an explanation for the positive relationship between chronic hyperinsulinaemia, hepatic VLDL triacylglycerol secretion and hypertriglyceridaemia observed previously in insulin-resistant states. PMID- 10672000 TI - Regiospecific glycosidase-assisted synthesis of lacto-N-biose I (Galbeta1 3GlcNAc) and 3'-sialyl-lacto-N-biose I (NeuAcalpha2-3Galbeta1-3GlcNAc). AB - The all-transglycolytic synthesis of lacto-N-biose I (Galbeta1-3GlcNAc) and 3' sialyl-lacto-N-biose I (NeuAcalpha2-3Galbeta1-3GlcNAc) was performed. The disaccharide lacto-N-biose I was obtained by use of p-nitrophenyl beta-D galactopyranoside as the donor, 2-acetamido-2-deoxy-D-glucopyranose as the acceptor and Xanthomonas manihotis beta-D-galactosidase as the catalyst. The reaction was shown to be regiospecific, with a high molar yield (about 55%) with respect to the donor. Lacto-N-biose I obtained by this method was used as the acceptor for a subsequent enzymatic reaction catalyzed by Trypanosoma cruzi trans sialidase in which 2'-(4-methylumbellyferyl)-alpha-D-N-acetylneuraminic was used as the donor of the N-acetylneuraminil moiety. The reaction generated the product, 3'-sialyl-lacto-N-biose I, regiospecifically and with a molar yield of about 35%. PMID- 10672001 TI - Electron transfer rates and equilibrium within cytochrome c oxidase. AB - Intramolecular electron transfer (ET) between the CuA center and heme a in bovine cytochrome c oxidase was investigated by pulse radiolysis. CuA, the initial electron acceptor, was reduced by 1-methyl nicotinamide radicals in a diffusion controlled reaction, as monitored by absorption changes at 830 nm. After the initial reduction phase, the 830 nm absorption was partially restored, corresponding to reoxidation of the CuA center. Concomitantly, the absorption at 445 nm and 605 nm increased, indicating reduction of heme a. The rate constants for heme a reduction and CuA reoxidation were identical within experimental error and independent of the enzyme concentration. This demonstrates that a fast intramolecular electron equilibration is taking place between CuA and heme a. The rate constants for CuA --> heme a ET and the reverse (heme a --> CuA) process were found to be 13 000 s-1 and 3700 s-1, respectively, at 25 degrees C and pH 7.4. This corresponds to an equilibrium constant of 3.4 under these conditions. Thermodynamic and activation parameters of the ET reactions were determined. The significance of these results, particularly the observed low activation barriers, are discussed within the framework of the known three-dimensional structure, ET pathways and reorganization energies. PMID- 10672002 TI - Insulin stimulates NHE1 activity by sequential activation of phosphatidylinositol 3-kinase and protein kinase C zeta in human erythrocytes. AB - The signaling cascade linking insulin receptor stimulation to the activation of Na/H exchanger (NHE) was investigated in human erythrocytes, a simple cell model expressing the NHE1 isoform and protein kinase C (PKC) alpha and zeta isoforms only. Our results demonstrate the presence of phosphatidylinositol (PtdIns) 3 kinase in these cells and its activation by insulin. With a similar time-course, insulin also promoted both the translocation and activation of PKC zeta, but had no effect on PKC alpha. Inhibition of PtdIns 3-kinase with wortmannin prevented the activation of PKC zeta by insulin. Stimulation of NHE1 was observed after 10 min of insulin treatment and persisted for at least 60 min. This effect was totally abolished by wortmannin or GF 109203X, an inhibitor of all PKC isoforms, but not by Go 6976, a specific inhibitor of conventional and novel PKCs (e.g. PKC alpha). These data indicate that PKC zeta activation is mediated by a PtdIns 3 kinase-dependent mechanism and that NHE1 stimulation involves the sequential activation of PtdIns 3-kinase and PKC zeta. In addition, insulin stimulation of NHE1 occurred without altering the phosphorylation state of the exchanger, suggesting that the phosphorylation of an ancillary protein by PKC zeta would be responsible for activation of the transporter. PMID- 10672003 TI - Regulation of phenobarbital induction of the cytochrome P450 2b9/10 genes in primary mouse hepatocyte culture. Involvement of calcium- and cAMP-dependent pathways. AB - Phenobarbital (PB) has long been known as an inducer of drug-metabolizing enzymes in liver, but the molecular mechanism underlying this induction is still poorly understood. Using primary mouse hepatocyte culture, we have investigated the possible involvement of different regulatory pathways in PB action, by exposing PB-treated cells to various protein kinase/phosphatase modulators. Our results showed a negative role of the cAMP-dependent pathway, as treatment with cAMP dependent protein kinase (PKA) activators (10 microM dibutyryl-cAMP and 50 microM forskolin) dramatically inhibited PB-induced Cyp2b9/10 mRNA accumulation, whereas PKA inhibitor potentiated the PB responsiveness of this gene. The cGMP-dependent protein kinase (PKG) seems to play a positive role as PKG inhibitor reduced the PB-induced level of Cyp2b9/10 mRNA. We also obtained two lines of evidence for the involvement of Ca2+ in modulating PB action. Firstly, measurements of intracellular Fura-2 fluorescence ratio in murine hepatocytes showed that long term PB incubation (24 and 48 h) led to a significant increase of [Ca2+]i. Secondly, treatment with an intracellular Ca2+ chelator (BAPTA-AM) nearly completely abolished PB-induced Cyp2b9/10 expression. Ca2+ thus appeared to mediate PB action likely via Ca2+/calmodulin-dependent protein kinase II, as KN62, a specific inhibitor of this enzyme, also dramatically inhibited PB induction of the Cyp2b9/10 genes. PMID- 10672004 TI - Transforming growth factor beta induction of insulin gene expression is mediated by pancreatic and duodenal homeobox gene-1 in rat insulinoma cells. AB - Although transforming growth factor-beta (TGF-beta) stimulates pancreatic islet cells to synthesize and secret insulin, the mechanism underlying this effect is not known. To investigate this question, we examined the insulin promoter activity focusing on a transcription factor, pancreatic and duodenal homeobox gene-1 (PDX-1) that binds to the A3 element of the rat insulin promoter. Studies performed using the rat insulinoma cell line, INS-1 showed that TGF-beta stimulation of endogenous insulin mRNA expression correlated with increased activity of a reporter construct containing the insulin promoter. A potential mechanism for this increase arose from, electrophoretic mobility shift assay showing that the nuclear extract from TGF-beta treated cells contained higher levels of A3 binding activity. Western blot analysis confirmed that PDX-1 was increased in the nuclear extract from INS-1 cells treated with TGF-beta. As expected, a mutant insulin promoter that lacked the PDX-1 binding site was not stimulated by TGF-beta. In summary, the results of these studies show that TGF beta stimulates the transcription of insulin gene and this action is mediated by the transcription factor, PDX-1. PMID- 10672005 TI - Activity and stability of a neutral protease from Vibrio sp. (vimelysin) in a pressure-temperature gradient. AB - The apparent second-order rate constant of hydrolysis of Fua-Gly-LeuNH2 by vimelysin, a neutral protease from Vibrio sp. T1800, was measured in a variable pressure-temperature gradient (0. 1-400 MPa and 5-40 degrees C). The apparent maximum rate was observed at approximately 15 degrees C and 150-200 MPa; the pressure-activation ratio (kcat/Km(max)/kcat/Km(0.1 MPa)) was reached about sevenfold. The pressure dependence of the kcat and Km parameters at constant temperature (25 degrees C) revealed that the pressure-activation below 200 MPa was mainly caused by a change in the kcat parameter. The change in the intrinsic fluorescence intensity of vimelysin was also measured in a pressure-temperature plane (0.1-400 MPa and -20 to +60 degrees C). The fluorescence intensity was found to decrease by increasing pressure and temperature, and the isointensity contours were more or less circular. The tangential lines to the contours at high temperatures and low to medium pressures seem to have slightly positive slopes, which was reflected by the higher residual activities left after incubations at higher temperatures and medium pressure (200 MPa and 50 degrees C) and by the almost intact secondary structure left after 1 h of incubation at 200 MPa and 40 degrees C, as studied by circular dichroism. These results were compared with the corresponding results for thermolysin, a moderately thermostable protease from Bacillus thermoproteolyticus. Apparent differences that might be related to the temperature adaptations of the respective source microbes are also discussed. PMID- 10672006 TI - Inhibition of phosphatidylserine synthesis during Jurkat T cell activation. The phosphatase inhibitor, sodium ortho-vanadate bypasses the CD3/T cell receptor induced second messenger signaling pathway. AB - Sodium ortho-vanadate (Na3VO4), an inhibitor of protein tyrosine phosphatase, induces a rapid (15 min) and strong inhibition of phosphatidylserine synthesis with an IC50 = 100 microM. The mode of action of Na3VO4 was compared to that of CD3 mAbs. It was found that Na3VO4 bypasses the major CD3-induced T cell activation signals including protein tyrosine phosphorylation, p56lck activation and the generation of second messengers including inositol phosphates and its subsequent Ca2+ mobilization as well as diacylglycerol production. These facts were confirmed by using a panel of Jurkat clones that differs by the expression of either tyrosine kinases involved in the CD3-induced T cell activation pathway such as p56lck, p72syk and ZAP-70 or some cell surface receptors such as the CD3/TCR complex or the CD45 phosphatase. PMID- 10672007 TI - Sulfite and membrane energization induce two different active states of the Paracoccus denitrificans F0F1-ATPase. AB - Activation of the latent ATPase activity of inside-out vesicles from plasma membranes of Paracoccus denitrificans was studied. Several factors were found to induce activation: heat, membrane energization by succinate oxidation, methanol, oxyanions (sulfite, phosphate, arsenate, bicarbonate) and limited proteolysis with trypsin. Among the oxyanions, sulfite induced the higher increase in ATPase activity. Sulfite functioned as a nonessential activator that slightly modified the affinity for ATP and increased notoriously the Vmax. There was a competitive effect between sulfite, bicarbonate and phosphate for ATPase activation; their similar chemical geometry suggests that these oxyanions have a common binding site on the enzyme. Dithiothreitol did not affect the ATPase activity. ATPase activation by sulfite was decreased by uncoupler, enhanced by trypsin and inhibited by ADP, oligomycin and venturicidin. In contrast, activation induced by succinate was less sensitive to ADP, oligomycin, venturicidin and trypsin. It is proposed that the active states induced by sulfite and succinate reflect two conformations of the enzyme, in which the inhibitory subunit epsilon is differently exposed to trypsin. PMID- 10672008 TI - Dual regulatory effects of nitric oxide on plasminogen activator inhibitor type 1 expression in endothelial cells. AB - In this report we compared the mechanism by which nitric oxide (NO), generated exogenously and endogenously, affects the plasminogen activator inhibitor type 1 (PAI-1) expression in endothelial cells. For this purpose, we stimulated the endothelial cell line EA.hy 926 with tumour necrosis factor alpha (TNFalpha) in the presence of the exogenously NO-releasing donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine, or regulators of nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine-methyl ester hydrochloride and substrate L-Arg. Expression of PAI-1 in EA.hy 926 cells was determined by measuring the level of mRNA, using relative quantitative reverse transcriptase PCR, and protein, using ELISA. In addition, we estimated the level of activation of two mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK1/2), in the cells before and after treatment with TNFalpha, in the presence or absence of NO donors and inhibitors. In contrast to exogenously released NO that significantly reduced mostly basal PAI-1 expression, endogenously generated NO by NOS potentiated TNFalpha-induced upregulation of PAI 1 expression. Exogenously and endogenously generated NO causes different effects on activation of the MAPKs ERK1/2 and JNK1/2. Specifically, the SNP-released NO activates only ERK1/2, while endogenously generated NO in a pathway induced by TNFalpha activates both MAPKs. Thus our data indicate that due to different cellular locations and mechanisms of generation, NO may participate in various signalling pathways leading to opposite effects on PAI-1 expression in endothelial cells. PMID- 10672009 TI - High resolution solution structure of the protein part of Cu7 metallothionein. AB - The three-dimensional solution structure of the protein part of Cu7 metallothionein (Cu7MT) of Saccharomyces cerevisiae has been attempted by 1H two dimensional NMR spectroscopy at 800 MHz. The protein part constitutes 53 amino acids. A total of 1192 NOEs, of which 1048 are meaningful, were used to determine the solution structure of the first 40 residues, the last 13 residues being disordered. A family of 30 structures was generated. Root-mean-square deviation (rmsd) values from the average structure of 0.32 +/- 0.13 A and 0.61 +/- 0.15 A for backbone and all heavy atoms, respectively, were obtained for the residues 2 40. The ten copper-coordinating cysteine sulfurs and the empty spaces around them are well defined. The structure of the protein part is similar but not identical to the available ones of the same holoprotein and of the Ag7 metallothionein, and is qualitatively superior. If the same metal-sulfur connectivities reported in the literature from 1H-109Ag heteronuclear multiple quantum coherence spectroscopy are assumed to hold for the present copper derivative, a peptide structure is obtained which is again similar, but still not identical, within indetermination, to that available. The structure of the copper polymetallic center may well be different from that proposed for the silver derivative, and indeed a number of different arrangements of the seven copper ions are consistent with the present highly refined structure of the protein part. PMID- 10672010 TI - Specific inhibition of barley alpha-amylase 2 by barley alpha-amylase/subtilisin inhibitor depends on charge interactions and can be conferred to isozyme 1 by mutation. AB - alpha-Amylase 2 (AMY2) and alpha-amylase/subtilisin inhibitor (BASI) from barley bind with Ki = 0.22 nM. AMY2 is a (beta/alpha)8-barrel enzyme and the segment Leu116-Phe143 in domain B (Val89-Ile152), protruding at beta-strand 3 of the (beta/alpha)8-barrel, was shown using isozyme hybrids to be crucial for the specificity of the inhibitor for AMY2. In the AMY2-BASI crystal structure [F. Vallee, A. Kadziola, Y. Bourne, M. Juy, K. W. Rodenburg, B. Svensson & R. Haser (1998) Structure 6, 649-659] Arg128AMY2 forms a hydrogen bond with Ser77BASI, while Asp142AMY2 makes a salt-bridge with Lys140BASI. These two enzyme residues are substituted by glutamine and asparagine, respectively, to assess their contribution in binding of the inhibitor. These mutations were performed in the well-expressed, inhibitor-sensitive hybrid barley alpha-amylase 1 (AMY1)-(1 90)/AMY2-(90-403) with Ki = 0.33 nM, because of poor production of AMY2 in yeast. In addition Arg128, only found in AMY2, was introduced into an AMY1 context by the mutation T129R/K130P in the inhibitor-insensitive hybrid AMY1-(1-161)/AMY2 (161-403). The binding energy was reduced by 2.7-3.0 kcal.mol-1 as determined from Ki after the mutations R128Q and D142N. This corresponds to loss of a charged interaction between the protein molecules. In contrast, sensitivity to the inhibitor was gained (Ki = 7 microM) by the mutation T129R/K130P in the insensitive isozyme hybrid. Charge screening raised Ki 14-20-fold for this latter mutant, AMY2, and the sensitive isozyme hybrid, but only twofold for the R128Q and D142N mutants. Thus electrostatic stabilization was effectively introduced and lost in the different mutant enzyme-inhibitor complexes and rational engineering using an inhibitor recognition motif to confer binding to the inhibitor mimicking the natural AMY2-BASI complex. PMID- 10672011 TI - A class of zinc fingers involved in protein-protein interactions biophysical characterization of CCHC fingers from fog and U-shaped. AB - Zinc fingers (ZnFs) are extremely common protein domains. Several classes of ZnFs are distinguished by the nature and spacing of their zinc-coordinating residues. While the structure and function of some ZnFs are well characterized, many others have been identified only through their amino acid sequence. A number of proteins contain a conserved C-X2-C-X12-H-X1-5-C sequence, which is similar to the spacing observed for the 'classic' CCHH ZnFs. Although these domains have been implicated in protein-protein (and not protein-nucleic acid) interactions, nothing is known about their structure or function at a molecular level. Here, we address this problem through the expression and biophysical characterization of several CCHC type zinc fingers from the erythroid transcription factor FOG and the related Drosophila protein U-shaped. Each of these domains does indeed fold in a zinc dependent fashion, coordinating the metal in a tetrahedral manner through the sidechains of one histidine and three cysteine residues, and forming extremely thermostable structures. Analysis of CD spectra suggests an overall fold similar to that of the CCHH fingers, and indeed a point mutant of FOG-F1 in which the final cysteine residue is replaced by histidine remains capable of folding. However, the CCHC (as opposed to CCHH) motif is a prerequisite for GATA-1 binding activity, demonstrating that CCHC and CCHH topologies are not interchangeable. This demonstration that members of a structurally distinct subclass of genuine zinc finger domains are involved in the mediation of protein-protein interactions has implications for the prediction of protein function from nucleotide sequences. PMID- 10672012 TI - Structural comparison of psychrophilic and mesophilic trypsins. Elucidating the molecular basis of cold-adaptation. AB - Structural rationalizations for differences in catalytic efficiency and stability between mesophilic and cold-adapted trypsins have been suggested from a detailed comparison of eight trypsin structures. Two trypsins, from Antarctic fish and Atlantic cod, have been constructed by homology modeling techniques and compared with six existing X-ray structures of both cold-adapted and mesophilic trypsins. The structural analysis focuses on the cold trypsin residue determinants found in a more extensive comparison of 27 trypsin sequences, and reveals a number of structural features unique to the cold-adapted trypsins. The increased substrate affinity of the psychrophilic trypsins is probably achieved by a lower electrostatic potential of the S1 binding pocket particularly arising from Glu221B, and from the lack of five hydrogen bonds adjacent to the catalytic triad. The reduced stability of the cold trypsins is expected to arise from reduced packing in two distinct core regions, fewer interdomain hydrogen bonds and from a destabilized C-terminal alpha-helix. The helices of the cold trypsins lack four hydrogen bonds and two salt-bridges, and they have poorer van der Waals packing interactions to the body of the molecule, compared to the mesophilic counterparts. PMID- 10672013 TI - Resonance Raman study of multihemic c-type cytochromes from Desulfuromonas acetoxidans. AB - Two multihemic cytochromes c from the sulfur reducing bacteria Desulfuromonas acetoxidans have been studied by optical and resonance Raman spectroscopy: cytochrome c551.5, a trihemic cytochrome and cytochrome c Mr 50 000, a recently isolated high molecular mass cytochrome. The redox and Raman characteristics of cytochrome c551.5 are compared to those of the tetrahemic cytochromes c3 from Desulfovibrio. While the redox behavior, followed by spectroelectrochemistry, is similar to that of cytochrome c3, showing the same conformational change after reduction of the highest potential heme, the Raman data show a contribution from a His- form of the axial ligands and lead to the assignment of a band at 218 cm-1 to the Fe(III)-(His)2 stretching vibration. The Raman data on cytochrome c Mr 50 000 are in favor of an entirely low spin species with two different sets of axial ligands. A partially reduced state is easily accessible by ascorbate addition. PMID- 10672014 TI - Complementation of NADPH oxidase in p67-phox-deficient CGD patients p67-phox/p40 phox interaction. AB - Chronic granulomatous disease (CGD) is due to a functional defect of the O2- generating NADPH oxidase of phagocytes. Epstein-Barr-virus-immortalized B lymphocytes express all the constituents of oxidase with activity 100 times less than that of neutrophils. As in neutrophils, oxidase activity of Epstein-Barr virus-immortalized B lymphocytes was shown to be defective in the different forms of CGD; these cells were used as a model for the complementation studies of two p67-phox-deficient CGD patients. Reconstitution of oxidase activity was performed in vitro by using a heterologous cell-free assay consisting of membrane-suspended or solubilized and purified cytochrome b558 that was associated with cytosol or with the isolated cytosolic-activating factors (p67-phox, p47-phox, p40-phox) from healthy or CGD patients. In p67-phox-deficient CGD patients, two cytosolic factors are deficient or missing: p67-phox and p40-phox. Not more than 20% of oxidase activity was recovered by complementing the cytosol of p67-phox-deficient patients with recombinant p67-phox. On the contrary, a complete restoration of oxidase activity was observed when, instead of cytosol, the cytosolic factors were added in the cell-free assay after isolation in combination with cytochrome b558 purified from neutrophil membrane. Moreover, the simultaneous addition of recombinant p67-phox and recombinant p40-phox reversed the previous complementation in a p40-phox dose-dependent process. These results suggest that in the reconstitution of oxidase activity, p67-phox is the limiting factor; the efficiency of complementation depends on the membrane tissue and the cytosolic environment. In vitro, the transition from the resting to the activated state of oxidase, which results from assembling, requires the dissociation of p40-phox from p67-phox for efficient oxidase activity. In the process, p40-phox could function as a negative regulatory factor and stabilize the resting state. PMID- 10672015 TI - Cytochrome cM from synechocystis 6803. Detection in cells, expression in Escherichia coli, purification and physical characterization. AB - Based on DNA sequence data a novel c-type cytochrome, cytochrome cM, has been predicted to exist in the cyanobacterium Synechocystis 6803. The precursor protein consists of 105 amino acids with a characteristic heme-binding motif and a hydrophobic domain located at the N-terminal end that is proposed to act as either a signal peptide or a membrane anchor. For the first time we report the detection of cytochrome cM in Synechocystis 6803 using Western blot analysis. The soluble portion cytochrome cM has been overexpressed in Escherichia coli in two forms, one with a poly histidine tag to facilitate purification and one without such a tag. The overexpressed protein has been purified and shown to bind heme, exhibiting an absorption peak in the Soret band near 416 nm and a peak in the alpha band at 550 nm. The extinction coefficient of cytochrome cM is 23.2 +/- 0.5 mM-1.cm-1 for the reduced minus oxidized alpha band peak (550-535 nm). The isoelectric point of cytochrome cM is 5.6 (without the histidine tag), which is significantly lower than the pI of 7.2 predicted from the amino acid sequence. The redox midpoint potential of cytochrome cM expressed in E. coli is 151 +/- 5 mV (pH 7.1), which is quite low compared to other c-type cytochromes in which a histidine and a methionine residue serve as the axial ligands to the heme. This work opens the way for determining the three-dimensional structure of cytochrome cM and investigating its function in cyanobacteria. PMID- 10672016 TI - Contribution of Are1p and Are2p to steryl ester synthesis in the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, two acyl-CoA:sterol acyltransferases (ASATs) that catalyze the synthesis of steryl esters have been identified, namely Are2p (Sat1p) and Are1p (Sat2p). Deletion of either ARE1 or ARE2 has no effect on cell viability, and are1are2 double mutants grow in a similar manner to wild-type despite the complete lack of cellular ASAT activity and steryl ester formation [Yang, H., Bard, M., Bruner, D. A., Gleeson, A., Deckelbaum, R. J., Aljinovic, G., Pohl, T. M., Rothstein, R. & Sturley, S. L. (1996) Science 272, 1353-1356; Yu, C., Kennedy, J., Chang, C. C. Y. & Rothblatt, J. A. (1996) J. Biol. Chem. 271, 24157-24163]. Here we show that both Are2p and Are1p reside in the endoplasmic reticulum as demonstrated by measuring ASAT activity in subcellular fractions of are1 and are2 deletion strains. This localization was confirmed by fluorescence microscopy using hybrid proteins of Are2p and Are1p fused to green fluorescent protein (GFP). Lipid analysis of are1 and are2 deletion strains revealed that Are2p and Are1p utilize sterol substrates in vivo with different efficiency; Are2p has a significant preference for ergosterol as a substrate, whereas Are1p esterifies sterol precursors, mainly lanosterol, as well as ergosterol. The specificity towards fatty acids is similar for both isoenzymes. The lack of steryl esters in are1are2 mutant cells is largely compensated by an increased level of free sterols. Nevertheless, terbinafine, an inhibitor of ergosterol biosynthesis, inhibits growth of are1are2 cells more efficiently than growth of wild-type. In a growth competition experiment are1are2 cells grow more slowly than wild-type after several rounds of cultivation, suggesting that Are1p and Are2p or steryl esters, the product formed by these two enzymes, are more important in the natural environment than under laboratory conditions. PMID- 10672017 TI - Changes in integrity and association of eukaryotic protein synthesis initiation factors during apoptosis. AB - Induction of apoptosis results in inhibition of the rate of overall protein synthesis in a variety of cell types. We have shown previously that polypeptide chain initiation factor eIF4GI is rapidly cleaved by caspase-3, whereas other components of the eIF4F complex are much more stable during apoptosis in BJAB and Jurkat cells. We have now extended our analysis to other factors involved in the initiation of protein synthesis and we report here that eIF4B, the p35 subunit of eIF3, and minor proportions of the alpha subunit of eIF2 and the eIF4E-binding protein 4E-BP1 are also cleaved to give rise to discrete fragments. These cleavages occur with delayed kinetics relative to that seen for eIF4GI, and eIF2beta and eIF2gamma levels also decrease at a relatively late stage of apoptosis. In contrast, the second form of eIF4G described recently, eIF4GII, is cleaved as rapidly as eIF4GI under the same conditions. Purified recombinant caspase-3 is able to degrade eIF4B and eIF3(p35) in vitro, producing fragments of the same sizes as those seen in intact cells. Induction of apoptosis also results in a biphasic change in the association of 4E-BP1 with eIF4E. Thus the progress of apoptosis is characterized by a complex programme of changes in several initiation factors, including the specific fragmentation or complete degradation of some and alterations in the association status of others. These events are likely to contribute to the inhibition of protein synthesis seen under these conditions. PMID- 10672018 TI - Functional characterization and mechanism of action of recombinant human kynurenine 3-hydroxylase. AB - The mitochondrial outer membrane enzyme kynurenine 3-hydroxylase (K3H) is an NADPH-dependent flavin mono-oxygenase involved in the tryptophan pathway, where it catalyzes the hydroxylation of kynurenine. K3H was transiently expressed in COS-1 cells as a glutathione S-transferase (GST) fusion protein, and the pure recombinant protein (rec-K3H) was obtained with a specific activity of about 2000 nmol.min-1.mg-1. Rec-K3H was shown to have an optimum pH at 7.5, to use NADPH more efficiently than NADH, and to contain one molecule of non-covalently bound FAD per molecule of enzyme. The mechanism of the rec-K3H-catalyzed reaction was investigated by overall initial-rate measurements, and a random mechanism in which combination of the enzyme with one substrate does not influence its affinity for the other is proposed. Further kinetic studies revealed that K3H activity was inhibited by both pyridoxal phosphate and Cl-, and that NADPH catalyzed oxidation occurred even in the absence of kynurenine if 3 hydroxykynurenine was present, suggesting an uncoupling effect of 3 hydroxykynurenine with peroxide formation. This observation could be of clinical interest, as peroxide formation could explain the neurotoxicity of 3 hydroxykynurenine in vivo. PMID- 10672019 TI - The N-terminal beta-barrel structure of lipid body lipoxygenase mediates its binding to liposomes and lipid bodies. AB - Phospholipase A2 and a particular isoform of lipoxygenase are synthesized and transferred to lipid bodies during the stage of triacylglycerol mobilization in germinating cucumber seedlings. Lipid body lipoxygenase (LBLOX) is post translationally transported to lipid bodies without proteolytic modification. Fractionation of homogenates from cucumber cotyledons or transgenic tobacco leaves expressing LBLOX showed that a small but significant amount was detectable in the microsomal fraction. A beta-barrel-forming N-terminal domain in the structure of LBLOX, as deduced from sequence data, was shown to be crucial for selective intracellular transport from the cytosol to lipid bodies. Although a specific signal sequence for targeting protein domains to the lipid bodies could not be established, it was evident that the beta-barrel represents a membrane binding domain that is functionally comparable with the C2 domains of mammalian phospholipases. The intact beta-barrel of LBLOX was demonstrated to be sufficient to target in vitro a fusion protein of LBLOX beta-barrel with glutathione S transferase (GST) to lipid bodies. In addition, binding experiments on liposomes using lipoxygenase isoforms, LBLOX deletions and the GST-fusion protein confirmed the role of the beta-barrel as the membrane-targeting domain. In this respect, the cucumber LBLOX differs from cytosolic isoforms in cucumber and from the soybean LOX-1. When the beta-barrel of LBLOX was destroyed by insertion of an additional peptide sequence, its ability to target proteins to membranes was abolished. PMID- 10672020 TI - Characterization of hydroxylaminobenzene mutase from pNBZ139 cloned from Pseudomonas pseudoalcaligenes JS45. A highly associated SDS-stable enzyme catalyzing an intramolecular transfer of hydroxy groups. AB - Hydroxylaminobenzene mutase is the enzyme that converts intermediates formed during initial steps in the degradation of nitrobenzene to a novel ring-fission lower pathway in Pseudomonas pseudoalcaligenes JS45. The mutase catalyzes a rearrangement of hydroxylaminobenzene to 2-aminophenol. The mechanism of the reactions and the properties of the enzymes are unknown. In crude extracts, the hydroxylaminobenzene mutase was stable at SDS concentrations as high as 2%. A procedure including Hitrap-SP, Hitrap-Q and Cu(II)-chelating chromatography was used to partially purify the enzyme from an Escherichia coli clone. The partially purified enzyme was eluted in the void volume of a Superose-12 gel-filtration column even in the presence of 0.05% SDS in 25 mM Tris/HCl buffer, which indicated that it was highly associated. When the enzymatic conversion of hydroxylaminobenzene to 2-aminophenol was carried out in 18O-labeled water, the product did not contain 18O, as determined by GC-MS. The results indicate that the reaction proceeded by intramolecular transfer of the hydroxy group from the nitrogen to the C-2 position of the ring. The mechanism is clearly different from the intermolecular transfer of the hydroxy group in the non-enzymatic Bamberger rearrangement of hydroxylaminobenzene to 4-aminophenol and in the enzymatic hydroxymutation of chorismate to isochorismate. PMID- 10672021 TI - The wide binding properties of a wheat nonspecific lipid transfer protein. Solution structure of a complex with prostaglandin B2. AB - The 3D solution structure of wheat nonspecific lipid transfer protein (ns-LTP) complexed with prostaglandin B2, a lipid with both vinyl and hydroxylated groups, has been determined by 1H 2D NMR. The global fold of the protein is close to the previously published structures of wheat, maize, barley and rice ns-LTPs. The ligand is almost completely embedded in the hydrophobic core of the protein. Structure comparisons of free and bound wheat ns-LTP reveal that the binding of prostaglandin B2 hardly affects the global fold of the protein. The structural data on this unusual complex are discussed and compared with other known ns-LTP lipid-complexes. PMID- 10672022 TI - The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 degrees C. AB - The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 degrees C. It also showed high stability of 18.3 kcal.mol 1 (76.5 kJ.mol-1) at 25 degrees C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 degrees C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 degrees C. The cooperativity of ATP hydrolysis and ssDNA-binding ability of the protein above 75 degrees C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 degrees C being the critical threshold temperature. PMID- 10672023 TI - Insertion of light-harvesting chlorophyll a/b protein into the thylakoid topographical studies. AB - The major light-harvesting chlorophyll a/b-binding protein (Lhcb1,2) of photosystem II is inserted into the thylakoid via the signal recognition particle dependent pathway. However, the mechanism by which the protein enters the membrane is at this time unknown. In order to define some topographical restrictions for this process, we constructed several recombinant derivatives of Lhcb1 carrying hexahistidine tags at either protein terminus or in the stromal loop domain. Additionally, green fluorescent protein (GFP) was fused to either terminus. None of the modifications significantly impair the pigment-binding properties of the protein in the in vitro reconstitution of LHCII. With the exception of the C-terminal GFP fusion, all mutants stably insert into isolated thylakoids in the absence of Ni2+ ions. The addition of low concentrations of Ni2+ ions abolishes the thylakoid insertion of C-terminally His-tagged mutants whereas the other His-tagged proteins fail to insert only at higher Ni2+ concentrations. The C-terminus of Lhcb1 must cross the membrane during protein insertion whereas the other sites of Lhcb1 modification are positioned on the stromal side of LHCII. We conclude that a Ni2+-complexed His tag and fusion to GFP inhibit translocation of the protein C-terminus across the thylakoid. Our observations indicate that the N-terminal and stromal domain of Lhcb1 need not traverse the thylakoid during protein insertion and are consistent with a loop mechanism in which only the C-terminus and the lumenal loop of Lhcb1 are translocated across the thylakoid. PMID- 10672024 TI - Reciprocal relationship between alpha1,2 mannosidase processing and reglucosylation in the rough endoplasmic reticulum of Man-P-Dol deficient cells. AB - The study of the glycosylation pathway of a mannosylphosphoryldolichol-deficient CHO mutant cell line (B3F7) reveals that truncated Glc(0-3)Man5GlcNAc2 oligosaccharides are transferred onto nascent proteins. Pulse-chase experiments indicate that these newly synthesized glycoproteins are retained in intracellular compartments and converted to Man4GlcNAc2 species. In this paper, we demonstrate that the alpha1,2 mannosidase, which is involved in the processing of Man5GlcNAc2 into Man4GlcNAc2, is located in the rough endoplasmic reticulum. The enzyme was shown to be inhibited by kifunensine and deoxymannojirimycin, indicating that it is a class I mannosidase. In addition, Man4GlcNAc2 species were produced at the expense of Glc1Man5GlcNAc2 species. Thus, the trimming of Man5GlcNAc2 to Man4GlcNAc2, which is catalyzed by this mannosidase, could be involved in the control of the glucose-dependent folding pathway. PMID- 10672025 TI - Sulfakinin neuropeptides in a crustacean. Isolation, identification andtissue localization in the tiger prawn Penaeus monodon. AB - The sulfakinin (SK) family of neuropeptides are characterized by a C-terminal octapeptide sequence that begins with two acidic residues (most commonly DD), and ends with YGHMRF-NH2, usually with the tyrosyl residue sulfated. So far, sulfakinins have only been identified in insects and the present study was initiated to investigate if the family is more widely distributed within the arthropods. Purification of an extract of the central nervous system of the giant tiger prawn Penaeus monodon has revealed three novel members of the sulfakinin peptide family. One of the peptides, Pem SKI, has the sequence 95%) for a major clade of Pao-like retrotransposons, which includes five mosquito sequences and the recently discovered Drosophila retrotransposons BEL and ninja. This appears to represent a new family of Pao like LTR-retrotransposons distinct from the Copia and Gypsy families. PMID- 10672080 TI - Editorial PMID- 10672079 TI - Edgewalking to internationalize nursing. PMID- 10672081 TI - Best hope or last hope: access to phase III clinical trials of HER-2/neu for advanced stage breast cancer patients. AB - Breast cancer is a major public health problem, with a 12% incidence among women. The over-expression of the proto-oncogene HER-2/neu is associated with 30% of breast and ovarian cancers that are very aggressive and do not respond to standard therapeutic regimens. Entrance into clinical trials can represent the best hope and even the last hope for these patients. Entrance, however, is based on satisfying eligibility criteria. In examining advanced stage breast cancer patients' access to phase III clinical trials for HER-2/neu, two specific arguments regarding eligibility will be addressed. First, if research is to provide the utilitarian goal of the 'greatest good to the greatest number', delineation of the population receiving the 'good', rather than a homogeneous sub set of this population, must be addressed, along with patients' values and goals, very relevant to determining a 'good life', how to achieve it, and whether a treatment is a part of that process. Second, the 'good' being generated should involve realistic, practical values of quality ways of living with advanced breast cancer and not just increased survival, or cure. Arguments for relaxing criteria are based on an accurate versus over-simplified interpretation of utilitarian principles and concepts of human flourishing. Only through addressing these issues can the true 'good' of clinical trials and research be given to the greatest number of people. PMID- 10672082 TI - Equipoise: an appropriate standard for ethical review of nursing research? AB - When using control-group methodologies, the provision of different forms of treatment, which may vary in efficacy, to similar groups of research subjects must be ethically justified. The conventional justification is the existence of a state of clinical equipoise, defined as a lack of consensus among the expert community on the relative efficacy of the treatments. Control groups are justified when the profession does not know which treatment confers more benefit; otherwise, the provision of treatment known to be less effective than other available treatment is unethical. The concept of equipoise was developed to justify clinical trials of medical interventions. Equipoise has proven a durable standard in studies of interventions that confer benefit regardless of the subject's perception. However, equipoise does not apply when a substantive benefit is conferred by the subject's perception. In psychosocial interventions, a subject's experience of the intervention confers benefit and is used to evaluate benefit. Therefore the subject is in a valid position to prefer one treatment over the other and equipoise does not apply. PMID- 10672083 TI - Advanced nursing practice: the case of nurse practitioners in three Australian states. AB - This paper provides insight into the work of nurse practitioners in three Australian states. Using a case study approach, the aim of the study is to give an account of some of the types of cases/patients who consult with the nurse practitioners in the states visited and to offer insight into one of the policy changes required to support the introduction of the nurse practitioner role. A snowball sampling technique was used to obtain the sample population of both 10 nurse practitioners and other health care personnel. Semi-structured interviews were conducted with the 39 respondents in both samples. The analysis of the 10 nurse practitioner interviews reported here shows that these nurses undertook a wide range of roles. The results of the semi-structured interviews with the nurse practitioners are described using four of the 10 interviews and reporting them as case studies. The results highlighted the need to address legislation issues and to prepare nurses adequately for their clinical setting and address the difficulties of continuing education of rural nurses. PMID- 10672084 TI - Assessment of older people: politics and practice in primary care. AB - This paper reviews some of the key policy changes in the United Kingdom in relation to older people that have influenced assessment practice across health and social care, and have implications for nursing within emerging primary care groups. The role of assessment in purchasing care packages raises tensions and paradoxes for health and social care professionals that the paper sets out to elucidate with particular reference to district nurses. The issues for nursing have not been well described previously and questions such as key accountability, rationing, multiagency and interprofessional working, and the consumer voice, are discussed. PMID- 10672085 TI - Graduate nurses: critical thinkers or better decision makers? AB - This study evaluates the difference in development of critical thinking across four groups of nurses at different stages of the academic process and their perception of their decision-making ability in practice. With the move of nurse education into institutes of higher education nationally, there are no empirical data in the UK to suggest that graduates practice any differently from their non graduate colleagues. An opportunistic sample of 82 nurses, was chosen from recent admission on a pre-registration degree programme, to mature graduates, as well as a group of experienced, non-graduate practitioners. A quasi-experimental, between subjects design was used. A series of one-way ANOVAs was used to analyse the difference in critical thinking across all four groups, employing the Watson Glaser Critical Thinking Appraisal. Additionally, the Jenkins Clinical Decision Making in Nursing Scale was used to determine the differences in decision-making ability in practice across three of the groups with clinical experience. Furthermore, a correlation was undertaken to determine what relationship, if any, existed between critical thinking and decision-making in practice. It was found that there was no significant difference in the critical thinking skills across all groups studied, supporting the findings of other studies in the USA, which examined the cognitive skills of students undertaking graduate programmes. However, in their practice, it was found that those exposed to the academic process were significantly better at decision-making than their non-academic colleagues. Finally, no relationship could be found between the development of critical thinking and decision-making in practice, suggesting that more work needs to be done to look carefully at both critical thinking skills and decision making in practice and the tools used to measure these. PMID- 10672086 TI - Implementing contract learning in a clinical context: report on a study. AB - This paper reports on a study of the implementation and evaluation of learning contracts in a clinical context for a group of students who were in their third year of study in a pre-registration bachelor of nursing degree programme in Hong Kong. A learning contract was implemented as a learning tool in the students' clinical placement in mental health nursing. An action research approach was used in this study. Data were collected from questionnaires and interviews with students and clinical instructors. Results showed that students and clinical instructors generally agreed that there was an increase in students' autonomy and motivation in learning with the use of a learning contract. It also increased the sharing between students and clinical instructors. The findings of the questionnaire and interviews supported each other. However, the lack of experience in using contract learning and the limited time in the clinical areas created difficulties for both students and clinical instructors. Despite the limitations, contract learning is considered beneficial to students' learning and has the potential to be used in clinical learning. PMID- 10672087 TI - The minimization to clients of screen-detected breast cancer: a qualitative analysis. AB - Previous research has shown a low incidence of psychological morbidity in women with screen-detected breast cancer when compared to women with symptomatic breast cancer. Farmer et al. suggested that this might be due to the way the diagnosis of breast cancer is given to women with screen-detected disease. In order to test this hypothesis a detailed, in-depth, qualitative study was undertaken. The sample consisted of women with symptomatic breast cancer (n=5), women with screen detected invasive breast cancer (n=6) and women with screen-detected in-situ breast cancer (n=5). The 'bad news consultations' with the surgeons, and subsequent meetings with the breast care nurses (BCN), were tape recorded. The women were also interviewed in their own homes within 7 days. The results suggested that the women with screen-detected breast cancer received more reassurance than the women with symptomatic breast cancer and that the benefits of breast screening were emphasized by the surgeons and the BCNs. This led to minimization of the significance of screen-detected disease. Women were found to draw on a new conceptual model of early curable breast cancer which appears to be associated with a low incidence of psychological morbidity. PMID- 10672088 TI - How women receiving adjuvant chemotherapy for breast cancer cope with their treatment: a risk management perspective. AB - Retrospective interviews were undertaken with 12 women who had received an 18 week course of adjuvant chemotherapy for positive node breast cancer 1 year previously, and who had not experienced cancer recurrence. The nonstandardized interviews covered women's preconceptions about adjuvant chemotherapy, their information needs, and the impact of treatment. The qualitative data analysis drew upon the theoretical ideas of patient career, trajectory projection and qualitative risk analysis. Some women regarded adjuvant chemotherapy as no more than an 'insurance policy'. This perception may have arisen because doctors, attempting to minimize patient anxiety, did not discuss the high risk of disease recurrence which they faced. Other women equated adjuvant with curative chemotherapy, and anticipated hair loss or almost certain death. The women tried to cope with the physical and mental suffering associated with adjuvant chemotherapy through normalizing strategies, such as keeping a brave face, maintaining previous patterns of life, looking for humour and restructuring time. However, the rapid alterations in physical and mental state resulting from cycles of adjuvant chemotherapy resulted in a 'rollercoaster' experience for women which made normalization more difficult. Health professionals caring for women who must cope with uncertain future trajectories need to manage a risk communication dilemma. A strategy of fully informing women about the risks they face may cause anxiety or depression, and even impede recovery, given the evidence for psychological influences on health outcomes. But, if women do not understand the medical thinking on which their treatment is based, their misconceptions may be equally damaging. PMID- 10672089 TI - The patient's thoughts and feelings about their transfer from intensive care to the general ward. AB - This qualitative, phenomenological study was undertaken to find out how patients felt about being transferred from the intensive care unit to the ward. The six patients who took part in the study, had been in intensive care for more than 4 days. Their transfer was planned as part of their recovery from a critical illness. Following transfer to the ward, the patients were interviewed using a semi-structured approach. The interviews were audio-taped, and transcriptions from the tapes were analysed for meaning and interpretation. Findings supported the literature that was reviewed, in that transfer from intensive care to the ward was a traumatic experience for the patient, who often found it confusing, stressful and tiring. Improvements to the transfer process can be guided using the information from the literature review and findings of the study. PMID- 10672090 TI - Children visiting family and friends on adult intensive care units: the nurses' perspective. AB - Recent surveys show that children are still restricted from visiting their critically ill family and friends on many adult intensive care units throughout the country. The purpose of this small-scale exploratory pilot study was to examine and describe the experiences and perceptions of trained nurses towards children visiting within this setting. The aim of the study was to gain greater insight and understanding into the reason why, despite evidence to support the benefits to children of visiting their critically ill family and friends, they remain discouraged and restricted. It is hoped that the study will act as an initial enquiry to generate themes and further research questions. A qualitative research approach was adopted and in-depth focused interviews used as a method of data collection. The participants of the study were trained nurses working on an adult intensive care unit in a district general hospital in England. A total of 12 individual interviews were conducted which were audiotaped in full and analysed using a method of thematic content analysis. The value of the research is to promote family-centred care within an adult intensive care environment to meet the neglected needs of the well children of the critically ill person. The findings suggest that the participants in the study attempted to offer valuable support to children visiting their critically ill family and friends, but, despite an open visiting policy, children rarely visited within this setting. The desire of the well parent to protect and shield the child from the crisis of critical illness was perceived by the participants to be the main reason why they did not visit. To provide family-centred care within an adult intensive care setting has many implications for practice and several of these important issues are discussed. These include the educational and training needs of nursing staff and the importance of adopting a collaborative team approach to providing care for the critically ill person and their family. The need to generate research and literature from within the United Kingdom's health care system has also been identified and recommendations for further studies are proposed. PMID- 10672091 TI - The development of competency standards for specialist critical care nurses. AB - In defining the contemporary role of the specialist nurse it is necessary to challenge the concept of nursing as merely a combination of skills and knowledge. Nursing must be demonstrated and defined in the context of client care and include the broader notions of professional development and competence. This qualitative study sought to identify the competency standards for nurse specialists in critical care and to articulate the differences between entry-to practice standards and the advanced practice of specialist nurses. Over 800 hours of specialist critical care nursing practice were observed and grouped into 'domains' or major themes of specialist practice using a constant comparison qualitative technique. These domains were further refined to describe attributes of the registered nurses which resulted in effective and/or superior performance (competency standards) and to provide examples of performance (performance criteria) which met the defined standard. Constant comparison of the emerging domains, competency standards and performance criteria to observations of specialist critical care practice, ensured the results provided a true reflection of the specialist nursing role. Data analysis resulted in 20 competency standards grouped into six domains: professional practice, reflective practice, enabling, clinical problem solving, teamwork, and leadership. Each of these domains is comprised of between two and seven competency standards. Each standard is further divided into component parts or 'elements' and the elements are illustrated with performance criteria. The competency standards are currently being used in several Australian critical care educational programmes and are the foundation for an emerging critical care credentialling process. They have been viewed with interest by a variety of non-critical care specialty groups and may form a common precursor from which further specialist nursing practice assessment will evolve. PMID- 10672092 TI - Family members' experience of familial amyloidotic polyneuropathy disease--an infernal struggle and a fact of life. AB - Familial amyloidotic polyneuropathy is a fatal, hereditary, systemic, progressive amyloidosis. No previous qualitative study of the family members' experience of the disease has been published. The purpose of this phenomenological study was to understand the lived experience of family members whose nearest and dearest suffered from familial amyloidotic polyneuropathy. In-depth interviews were conducted with six family members. The analysis of the data was inspired by Colaizzi's method. Two major theme categories, difficult to accept and forced to accept, emerged from the interviews. Implications for nursing practice, such as genetic counselling and support, are discussed. PMID- 10672093 TI - Illness narratives of persons with post-polio syndrome. AB - This qualitative study investigated the lifetime illness experience of individuals with the 'late effects' of polio or post-polio syndrome. Fifteen individuals were interviewed twice about their illness experience and the interviews were transcribed verbatim. The empirical material first underwent a categorization process. The preliminary categories generated through this analysis were then condensed into broader categories which in the final analysis gave rise to the following temporal pattern or stages of the illness experience: (1) the acute phase of polio and subsequent treatment and care; (2) rehabilitation and care at institutions for the disabled; (3) adaptation to a new life; (4) living with the post-polio syndrome today, and finally, (5) memories of the past and apprehensions concerning the future. In spite of the difficult experiences of falling ill and slowly recovering from a life-threatening disease, these individuals have had a good life and accomplished most of their ambitions in the areas of work and family life. Their present psychosocial situation is complicated by the symptoms of the post-polio syndrome which make them more vulnerable to stress, but they are able to handle this burden except when any added strain makes it overwhelming. This potential vulnerability may sometimes express itself as a sudden flashback to traumatic polio experiences and it is therefore important that nurses are aware of the illness history of this patient group. PMID- 10672094 TI - Psychiatric nurses' thoughts and feelings about restraint use: a decision dilemma. AB - Patients continue to be physically restrained in psychiatric in-patient units. Studies concerned with staff-related variables have suggested that the emotional reactions of professionals to violent or potentially violent patients may influence their use of restrictive measures. However, no research existed that described psychiatric nurses' thoughts and feelings while they were involved in restraint situations nor what effects their thoughts and feelings had on their decision to restrain. Therefore, an ethnographic qualitative study was conducted in order to describe systematically nurses' thoughts and feelings toward restraint use in the in-patient psychiatric setting. The conceptual approach guiding the study was Etzioni's (1992) theoretical work on the role of normative affective factors in decision making. Following ethical approval of the study, ethnographic interviews were conducted with six nurses from an in-patient psychiatric unit who had participated in a situation involving the physical restraint of a patient. The analysis of the nurses' thoughts and feelings revealed that the restraint situation represented a decision dilemma for them. This overall finding was supported by four themes: (1) the framing of the situation: the potential for imminent harm; (2) the unsuccessful search for alternatives to physical restraints; (3) the conflicted nurse; and (4) the contextual conditions of restraint. The results indicated that restraint use is more complex than is currently conveyed in the literature in that normative affective factors influenced nurses' restraint decisions. The findings advance our understanding of why restraints continue to be used in psychiatric units. Further research is necessary to examine the findings in other settings and with a larger and more diverse population in order to draw definitive conclusions about the continued use of physical restraints in the care of patients on psychiatric units in hospitals. PMID- 10672095 TI - Patients' perceptions of seclusion: a qualitative investigation. AB - Twelve patients receiving acute in-patient psychiatric care in Queensland, Australia, participated in semi-structured interviews to elicit their perceptions of seclusion. All respondents had experienced time in seclusion within the 7 days prior to interview. Interviews were audiotaped, transcribed and analysed using content analysis. Five major themes emerged: use of seclusion, emotional impact, sensory deprivation, maintaining control and staff-patient interaction. The prevailing negativity towards seclusion underscores the need for ongoing critical review of its use. In particular, the relationship between patient responses to seclusion and the circumstances in which seclusion takes place requires greater consideration. Interventions such as providing information to patients about seclusion, increased interaction with patients during seclusion, attention to privacy and effective debriefing following seclusion may help to reduce the emotional impact of the practice. PMID- 10672096 TI - What do people need psychiatric and mental health nurses for? AB - The study reported here aimed to describe, by consulting with psychiatric practitioners of different disciplines, what people in contact with mental health services need nurses for, in terms of core nursing activity. Yet, recent trends have also been towards consumer-led definitions of good practice. The views of service and ex-service users can contribute much to an exploration of the role of psychiatric and mental health nurses and these perspectives were incorporated into the study. Given the lack of existing theory, a qualitative, grounded theory methodology was selected. In order to generate data rich enough for the analysis, focus groups of psychiatric nurses, social workers, service users, psychiatrists, carers and professions allied to medicine were sampled (13 groups, n=92) on the basis of the themes emerging from the data. Using critical incident technique (Flanagan 1954), the groups were invited to give examples of effective and ineffective nursing interventions, in relation to specific patient needs. The taped material was transcribed and analysed with the help of a computer package (QSR NUD.IST). This led to the selection of a core category, 'knowing you, knowing me', which described service users' and professionals' expectations that nurses are best placed to second guess the needs of patients and present themselves accordingly. Thus, nurses were expected, moment by moment, to know whether to be the patient's friend, a friendly professional, or take a more distant professional stance. The continuum entails different levels of knowledge and power, different language forms and different approaches to structuring time. Nurses themselves are most likely to prefer a position of friendly professional, from which they can move to a more intimate or distant role. Further study is needed to explore how nurses predict patients' expectations of them using a symbolic interactionist framework. PMID- 10672097 TI - Psychiatric nursing and compulsory psychiatric care. AB - Community psychiatric nurses (CPNs) have long been involved in the operations of the English Mental Health Acts. Research has shown that compulsory detention is not used uniformly or consistently. Rates of involuntary hospitalization are reported to vary widely across Europe, but there is some consensus on patient profiles. The ethnicity, social status and gender of the patient, the involvement of the police, the availability of care, problems caused to relatives, and the country and particular legislative system where these judgements take place, all influence who is compulsorily detained. This article reviews recent evidence from Europe and argues that involuntary psychiatric care can no longer be seen as entirely dependent on the symptoms and behaviour of the patient and that CPNs should be aware of and reflect upon these factors before invoking the detention process. PMID- 10672098 TI - Are patients who self-administer their medicines in hospital more satisfied with their care? AB - Patient self-administration of medicines in hospital prior to discharge is being seen increasingly as good practice by health professionals. Previous studies have looked for increased compliance and knowledge or asked whether patients liked self-administration and have not clearly demonstrated the benefits. This study surveys patients' views on self-administration and on their care. In particular it looks at the discharge process and the way information was given to the patient on discharge. Questionnaires were distributed to 309 patients being discharged from general medical wards of a teaching hospital in Central England to be completed by the patients after their discharge from hospital. Of these, 202 were returned to a separate university department for analysis. Although this study has been undertaken on only two wards, the survey has potential for large scale use. Findings which were consistent with previous work were that a great majority of patients who had self-administered their medicines would like to again, whilst those who had not had that opportunity were less likely to choose to self-administer in the future. However, this study found that a majority of patients under 60-years-old would choose to self-administer their medicines in hospital even if they had not been given the opportunity to do so recently. It was also found that patients who had administered their own medicines in hospital were more likely to report their overall care as excellent and were more satisfied with the discharge process than patients who had not. PMID- 10672099 TI - An evaluation of the safety and effectiveness of telephone triage as a method of patient prioritization in an ophthalmic accident and emergency service. AB - Service changes in the accident and emergency service at Manchester Royal Eye Hospital in England resulted in a telephone triage-based referral service for health care professionals. It became clear that this service needed evaluation in order to assure both providers and users of the service that this referral strategy, based on experienced nurse practitioners making decisions about patient priority, was safe and effective. The evaluation was extended to encompass the other area of the service where telephone referrals tend to be directly from patients. A mixed method was used. Information gained within the telephone triage conversation was compared with a final diagnosis through retrospective analysis of secondary data; the documentation of those patients who were not given access to the service initially was followed-up to ensure that this decision was safe and a number of nurses were questioned about telephone information gathering, using partially structured interviews. The study showed that nurse practitioners within the accident and emergency service were able to elicit accurate information from the telephone triage conversation on which to base a decision about patient access in most cases (76% over the whole service). This resulted in the appropriate prioritization of patients. No patient who needed urgent access to the service was denied it. The decisions made to deny urgent appointments to a number of patients were safe in all cases. It appears that one of the problem areas in the gathering of information for prioritization purposes is in the nurses' telephone triage discussions with some, but by no means all, doctors. Some general practitioners seemed unwilling to discuss the patient and give accurate information to a nurse and this is an area which appears to need some further work. PMID- 10672100 TI - Exploring the risks associated with induction of labour: a retrospective study using the NIMATS database. Northern Ireland Maternity System. AB - Induction of labour is a valuable obstetric procedure, providing obstetricians with the means to intervene should the health of the fetus be in jeopardy. Currently the most common reason for induction of labour is prolonged pregnancy, as obstetricians and midwives are concerned about the risks of postmaturity such as stillbirth, intrapartum asphyxia and birth trauma which are often associated with prolonged pregnancy (Lagrew & Freeman 1986). A retrospective comparative study was carried out in a large maternity unit to identify whether or not there was clinical evidence to support a policy of elective induction for post-term pregnancy. Three years' data were extracted from the Northern Ireland Maternity System (NIMATS) by writing new queries to the system. These data on 3262 women who delivered during 1994-96 were analysed to compare the outcomes for women who were induced with women who delivered spontaneously. Although the findings from the study in many instances failed to demonstrate statistical significance between the groups they did however, have important clinical significance. For example, those women who were induced had a 5% higher rate of caesarean section, 17% higher rate of epidural analgesia and on average a greater estimated blood loss. Statistical significance was evident when the apgar scores of the infants were compared; those induced had lower Apgars at 1 minute (7. 78 in the induced group compared to 7.9 in the spontaneous group [P < 0.01]) and at 5 min (8.99 in the induced group compared to 9.05 in the spontaneous group [P < 0.02]). PMID- 10672101 TI - Normal labour: a concept analysis. AB - Midwives practice within the normal childbirth paradigm. It is argued that midwives failure to define normality has allowed increasing technicalization and medicalization of the normal physiological process of birth because doctors so closely define abnormality. A concept analysis model is used to clarify what is meant by the term 'normal labour'; the emphasis being on understanding what normal labour is as it applies to midwifery practice today. This analysis highlights the importance of movement and the sequential nature of normal labour, and reveals how this is implicit within the other uses of both the words normal and labour. The final definition of normal labour offered is intended to be complimentary to existing medical determinants of progress of normal labour, because as the body of the text stresses, medical knowledge is fundamentally enmeshed in midwifery care. PMID- 10672102 TI - Critical thinking and nursing scripts: the case for the development of both. AB - It is argued that contemporary nursing education's emphasis on the conscious development of nurses' critical reasoning skills fails to take account of the complexity of both human cognition and clinical nursing practice. Human cognition centrally includes unconscious or tacit processes and clinical nursing practice is typified by the simultaneous presentation of clinical and non-clinical cues and competing clinical goals. Contemporary emphases on conscious critical thinking are largely consistent with the differing theoretical frameworks which have informed the study of nurses' clinical reasoning in the last 30 years, most of which permit the use of simulated case study. These frameworks, that is, decision theory, information processing and skills acquisition theory, are described and their limitations highlighted. In addition, an alternative theoretical framework, that of schema or script theory, which does take account of the complexities of cognition and practice, is discussed. Its implications for nursing practice and education are also outlined. PMID- 10672103 TI - Dialysis as 'deus ex machina': a critical analysis of haemodialysis. AB - This paper examines the context of dialysis from a critical and philosophical standpoint. The traditional attitude of dialysis as an all-saving medical wonder is examined. An alternative interpretation is offered using the analogy of 'experiment' to emphasize the uncertainty, power and control surrounding the treatment of end-stage renal disease. In order for the experiment (of dialysis) to occur, variables (patients) need to be controlled. Control can be found in the social structures around the experiment of dialysis which provide an environment conducive to its success. PMID- 10672104 TI - Cognitive behaviour therapy: teaching a client the ABC model--the first step towards the process of change. AB - Research suggests that clients need to be made aware of the relationship between thinking and emotional and behavioural reactions at the start of the therapeutic work. This paper looks at the clinical processes of educating the client about this relationship in a seven-step ABC model. Rationales for each of the seven steps are given. The model illustrates the process of generating alternative beliefs in relation to a negative event and shows consequences of holding helpful and unhelpful beliefs. PMID- 10672105 TI - Aggression and violence in health care professions. AB - Although violence is increasing in most workplaces, it has become a significant problem in health care professions. Not only has the number of incidents increased but also the severity of the impact has caused profound traumatic effects on the primary, secondary and tertiary victims. More health care professionals than ever are suffering from symptoms of post-traumatic stress disorder. Addressing the problem of violence in the workplace has been exacerbated by a lack of a clear definition of what constitutes aggression and violence. As a result, some administrators have been slow to commit resources to prevent further incidents and mitigate the impact. This article describes the magnitude of the problem from both an academic research and an operational perspective. A definition is presented as an initial step towards standardizing the research, and establishing an appropriate baseline upon which intervention policies and procedures can be created. This benchmark will also help to encourage empirical research into aggression and violence in health care professions and other professions. Further research needs to be conducted to create a comprehensive instrument that can more accurately measure the range of incidents and the severity of the impact. PMID- 10672106 TI - Educational input and patient outcomes: exploring the gap. AB - Over the last two decades, the health care professions in the United Kingdom have seen an unparalleled expansion of continuing professional education (CPE) and development (CPD) programmes; however, there is little empirical evidence that these enhance the care delivered to patients. Further research is also needed to demonstrate that these initiatives are linked to improved patient outcomes. If health care educators are to move towards an 'evidence-based curriculum', some restructuring of courses may be needed. Priorities should be set and decisions made, based on the results of reliable and valid research into the clinical outcomes of CPE. To evaluate courses and demonstrate educational effectiveness solely in terms of student satisfaction is not enough; to survive in the world of evidence-based care, educators must also demonstrate their contribution to clinical effectiveness. However, the neoteric field of impact evaluation lacks not only validated research methods, but also an agreed agenda for future research. Drawing on interviews undertaken with nurses who have participated in education evaluations and the relevant literature, this paper discusses the available data collection instruments and the development of viable research designs and methods, which are urgently needed to assess the outcomes of professional education programmes. PMID- 10672107 TI - Community development: a critical review of approaches to evaluation. AB - With the growing interest in community-based initiatives, this discussion paper focuses upon the evaluation of community development. It reviews three areas and highlights the complex and contentious nature of evaluation in this field. Commencing with approaches to evaluation, it critically reviews proposed methods and suggests that, rather than provide clarity and guidance, the competing designs are confusing for the potential researcher. Second, the discussion moves to evaluative terms that are applied to community development, suggesting that there is a mismatch between professional and lay interpretations with an ultimate impact upon the validity of their measurement. It uses the terms community and empowerment to make these differences more explicit. Finally, the paper argues that, although the aim of community-based initiatives is to bring about community health gain, gauging the pervasiveness of discreet project work is testing. It concludes by suggesting that a move away from an evaluative model based upon empowerment towards one which considers health and social capital may be a way forward. PMID- 10672108 TI - Validation of a new method for patient classification, the Oulu Patient Classification. AB - At Vasa Central Hospital in Western Finland a further development of the Oulu Patient Classification (OPC) has been made by the development of weight coefficients and by estimating the nursing care intensity per nurse. The daily level of nursing care intensity of a ward is expressed by the number of nursing care intensity points per nurse. This article presents results from a validity test of the OPC at Vasa Central Hospital. The test was carried out by comparing the daily patient classifications by means of the OPC against measurements made by means of a new measuring instrument, the 'Professional Assessment of Optimal Nursing Care Intensity Level' (PAONCIL) developed at the Vasa Central Hospital. The study was implemented in eight wards during a period of 3 months. The data material consisted of two parts, the daily patient classifications based on the OPC (n = 19 324) and the measurements by means of the PAONCIL forms (n = 8458). Simple and multiple linear regression analyses were used as statistical methods in quantifying the linear relationship between the two interval-scaled variables. In the test of concurrent validity the coefficient of determination was 0.366, i.e. the association between these two indicators is fairly strong (36.6%). The testing of construct validity showed that the construct validity of the indicator hardly deteriorates as a result of the patients being placed in separate nursing care intensity categories. There was a clear correlation between the scores allotted by the indicator to the six different sub-areas of nursing care. When examining the construct validity of the OPC, no factors with independent explanatory power in predicting PAONCIL values were discovered other than those of the OPC. The OPC proved on the basis of this research material and these statistical methods to possess fairly adequate validity, and thus there is a good basis for further research and a development of nursing care. PMID- 10672109 TI - Patient preferences for care by general internists and specialists in the ambulatory setting. AB - OBJECTIVE: To investigate patients' preferences for care by general internists and specialists for common medical conditions. DESIGN: Telephone interview. SETTING: A convenience sample of general internal medicine practices at 10 eastern academic medical centers. PATIENT/PARTICIPANTS: A probability sample of 314 participants who had at least one visit with their primary care physician during the preceding 2 years. MEASUREMENTS AND MAIN RESULTS: Items addressed patients' attitudes concerning continuity of care, preferences for care by general internists or specialists for common medical problems, and perceptions about the competency of general internists and specialists to manage these problems. Continuity was important to participants, with 63% reporting they preferred having one doctor. Respondents were willing to wait 3 or 4 days to see their regular doctor (85%) and wanted their doctor to see them in the emergency department (77%) and monitor their care while in the hospital (94%). A majority (>60%) preferred care from their regular doctor for a variety of new conditions. Though respondents valued continuity, 84% felt it was important to be able to seek medical care from any type of physician without a referral, and 74% responded that if they needed to see a specialist, they were willing to pay out of-pocket to do so. Although most participants (98%) thought their regular doctor was able to take care of usual medical problems, the majority thought that specialists were better able to care for allergies (79%) and better able to prescribe medications for depression (65%) and low-back pain (72%). CONCLUSIONS: Participants preferred to see their general internist despite their perceptions that specialists were more competent in caring for the conditions we examined. However, they wanted unrestricted access to specialists to supplement care provided by general internists. PMID- 10672110 TI - Screening and intervention for alcohol problems. A national survey of primary care physicians and psychiatrists. AB - OBJECTIVE: To describe adult primary care physicians' and psychiatrists' approach to alcohol screening and treatment, and to identify correlates of more optimal practices. DESIGN: Cross-sectional mailed survey. PARTICIPANTS: A national systematic sample of 2,000 physicians practicing general internal medicine, family medicine, obstetrics-gynecology, and psychiatry. MEASUREMENTS: Self reported frequency of screening new outpatients, and treatment recommendations in patients with diagnosed alcohol problems, on 5-point Likert-type scales. MAIN RESULTS: Of the 853 respondent physicians (adjusted response rate, 57%), 88% usually or always ask new outpatients about alcohol use. When evaluating patients who drink, 47% regularly inquire about maximum amounts on an occasion, and 13% use formal alcohol screening tools. Only 82% routinely offer intervention to diagnosed problem drinkers. Psychiatrists had the most optimal practices; more consistent screening and intervention was also associated with greater confidence in alcohol history taking, familiarity with expert guidelines, and less concern that patients will object. CONCLUSIONS: Most primary care physicians and psychiatrists ask patients about alcohol use, but fewer use recommended screening protocols or offer formal treatment. A substantial minority of physicians miss the opportunity to intervene in alcohol problems. Efforts to improve physicians' screening and intervention for alcohol problems should address their confidence in their skills, familiarity with expert recommendations, and beliefs that patients object to their involvement PMID- 10672111 TI - Bridging cultural differences in medical practice. The case of discussing negative information with Navajo patients. AB - BACKGROUND: Cultural differences between doctors and their patients are common and may have important implications for the clinical encounter. For example, some Navajo patients may regard advance care planning discussions to be a violation of their traditional values. OBJECTIVE: To learn from Navajo informants a culturally competent approach for discussing negative information. DESIGN: Focused ethnography. SETTING: Navajo Indian reservation, northeast Arizona. PARTICIPANTS: Thirty-four Navajo informants, including patients, traditional healers, and biomedical health care providers. MEASUREMENT: In-depth interviews. MAIN RESULTS: Strategies for discussing negative information were identified and organized into four stages. Assessment of patients is important because some Navajo patients may be troubled by discussing negative information, and others may be unwilling to have such discussions at all. Preparation entails cultivating a trusting relationship with patients, involving family members, warning patients about the nature of the discussion as well as communicating that no harm is intended, and facilitating the involvement of traditional healers. Communication should proceed in a caring, kind, and respectful manner, consistent with the Navajo concept k'e. Reference to a third party is suggested when discussing negative information, as is respecting the power of language in Navajo culture by framing discussions in a positive way. Follow-through involves continuing to care for patients and fostering hope. CONCLUSIONS: In-depth interviews identified many strategies for discussing negative information with Navajo patients. Future research could evaluate these recommendations. The approach described could be used to facilitate the bridging of cultural differences in other settings. PMID- 10672112 TI - Intensive care unit use and mortality in the elderly. AB - OBJECTIVE: To examine utilization and outcomes of intensive care unit (ICU) use for the elderly in the United States. DESIGN: We used 1992 data from the Health Care Financing Administration to examine ICU utilization and mortality by age and admission reason for hospitalizations of elderly Medicare beneficiaries. MAIN RESULTS: Use of the ICU was least likely for the oldest elderly overall (85+ years, 21.1% of admissions involved ICU; 75-84 years, 27.9%; 65-74 years, 29.7%), but more likely during surgical admissions. Eighty-three percent of the Medicare patients who received intensive care survived at least 90 days. Of the oldest elderly, 74% survived. Even among the 10% most expensive ICU hospitalizations, 77% of all patients and 62% of those 85 years and older survived at least 90 days. CONCLUSIONS: The likelihood of ICU use among these elderly decreased with age, especially among those 85 years or older. Diagnostic mix importantly influenced ICU use by age. The great majority of the elderly, including those 85 years and older and those receiving the most expensive ICU care, survived at least 90 days. PMID- 10672113 TI - The impact of leaving against medical advice on hospital resource utilization. AB - OBJECTIVE: To assess the effect of hospital discharge against medical advice (AMA) on the interpretation of charges and length of stay attributable to alcoholism. DESIGN: Retrospective cohort. Three analytic strategies assessed the effect of having an alcohol-related diagnosis (ARD) on risk-adjusted utilization in multivariate regressions. Strategy 1 did not adjust for leaving AMA, strategy 2 adjusted for leaving AMA, and strategy 3 restricted the sample by excluding AMA discharges. SETTING: Acute care hospitals. PATIENTS: We studied 23,198 pneumonia hospitalizations in a statewide administrative database. MEASUREMENTS AND MAIN RESULTS: Among these admissions, 3.6% had an ARD, and 1.2% left AMA. In strategy 1 an ARD accounted for a $1,293 increase in risk-adjusted charges for a hospitalization compared with cases without an ARD ( p =.012). ARD-attributable increases of $1,659 ( p =.002) and $1,664 ( p =. 002) in strategies 2 and 3 respectively, represent significant 28% and 29% increases compared with strategy 1. Similarly, using strategy 1 an ARD accounted for a 0.6-day increase in risk adjusted length of stay over cases without an ARD ( p =.188). An increase of 1 day was seen using both strategies 2 and 3 ( p =.044 and p =.027, respectively), representing significant 67% increases attributable to ARDs compared with strategy 1. CONCLUSIONS: Discharge AMA affects the interpretation of the relation between alcoholism and utilization. The ARD-attributable utilization was greater when analyses adjusted for or excluded AMA cases. Not accounting for leaving AMA resulted in an underestimation of the impact of alcoholism on resource utilization. PMID- 10672114 TI - Cost-effectiveness of low-molecular-weight heparin in the treatment of proximal deep vein thrombosis. AB - OBJECTIVE: To estimate the cost-effectiveness of low-molecular-weight heparin (LMWH) in the treatment of proximal lower extremity deep venous thrombosis. DESIGN: Cost-effectiveness analysis that includes the treatment of the index case and simulated 3-month follow-up. SETTING: Acute care facility. PATIENTS AND PARTICIPANTS: Hypothetical cohorts of 1,000 patients who present with proximal deep venous thrombosis. INTERVENTIONS: Intravenous unfractionated heparin (UH), LMWH (40% at home, 60% in hospital), or selective UH/LMWH (UH for hospitalized patients and LMWH for patients treated at home). MEASUREMENTS AND MAIN RESULTS: The outcomes were recurrent thrombosis, mortality, direct medical costs, and marginal cost-effectiveness ratios from the payer's perspective. At the base-case and under most assumptions in the sensitivity analysis, the LMWH and the selective UH/LMWH strategies dominate the UH strategy i.e., they result in fewer cases of recurrent thrombosis and fewer deaths, and they save resources. The savings occur primarily by decreasing the length of stay. The LMWH strategy resulted in lower costs as compared with the UH strategy when the proportion of patients treated at home was more than 14%. Treating 1, 000 patients with the LMWH strategy as compared with the UH/LMWH strategy would result in 10 fewer cases of recurrent thrombosis, 1.2 fewer deaths, at an additional cost of $96,822; the cost-effectiveness ratio was $9,667 and $80,685 per recurrent thrombosis or death prevented, respectively. CONCLUSIONS: Treatment with LMWH leads to savings and better outcomes as compared with UH in patients with lower extremity deep venous thrombosis. The selective UH/LMWH strategy is an alternative option. PMID- 10672115 TI - Health and health care among housestaff in four U.S. internal medicine residency programs. AB - BACKGROUND: Although there have been many studies of the health care services that resident physicians provide, little is known about the health care services they receive. OBJECTIVE: To describe residents' perceptions of the health care they receive. DESIGN: Anonymous mailed survey. SUBJECTS: All 389 residents in four U.S. categorical internal medicine training programs. MAIN RESULTS: Three hundred sixteen residents responded (83%). In aggregate, 116 (37%) reported having no primary care physician, and 36 (12%) reported that they are their own primary care physician. These figures varied substantially across the four programs. Most residents reported receiving basic screening and preventive services; however, their attitudes toward their health and health care differed across postgraduate level, gender, and program. Many residents reported that their long and unpredictable hours interfered with their ability to schedule clinician visits, that their health had declined because of residency, that programs and other residents were unsupportive of residents' health care needs, and that residency raised special issues of privacy that limited access to health care. CONCLUSIONS: Despite high rates of receipt of preventive services, these internal medicine residents identified several barriers that limited their access to health care. Program directors should explore these barriers and, at the same time, reevaluate the messages being sent to resident physicians about maintaining their health and health care. PMID- 10672116 TI - Is the professional satisfaction of general internists associated with patient satisfaction? AB - BACKGROUND: The growth of managed care has raised a number of concerns about patient and physician satisfaction. An association between physicians' professional satisfaction and the satisfaction of their patients could suggest new types of organizational interventions to improve the satisfaction of both. OBJECTIVE: To examine the relation between the satisfaction of general internists and their patients. DESIGN: Cross-sectional surveys of patients and physicians. SETTING: Eleven academically affiliated general internal medicine practices in the greater-Boston area. PARTICIPANTS: A random sample of English-speaking and Spanish-speaking patients (n = 2,620) with at least one visit to their physician (n = 166) during the preceding year. MEASUREMENTS: Patients' overall satisfaction with their health care, and their satisfaction with their most recent physician visit. MAIN RESULTS: After adjustment, the patients of physicians who rated themselves to be very or extremely satisfied with their work had higher scores for overall satisfaction with their health care (regression coefficient 2.10; 95% confidence interval 0.73-3.48), and for satisfaction with their most recent physician visit (regression coefficient 1.23; 95% confidence interval 0.26-2.21). In addition, younger patients, those with better overall health status, and those cared for by a physician who worked part-time were significantly more likely to report better satisfaction with both measures. Minority patients and those with managed care insurance also reported lower overall satisfaction. CONCLUSIONS: The patients of physicians who have higher professional satisfaction may themselves be more satisfied with their care. Further research will need to consider factors that may mediate the relation between patient and physician satisfaction. PMID- 10672117 TI - Evidence-based medicine training in internal medicine residency programs a national survey. AB - To characterize evidence-based medicine (EBM) curricula in internal medicine residency programs, a written survey was mailed to 417 program directors of U.S. internal medicine residency programs. For programs offering a freestanding (dedicated curricular time) EBM curriculum, the survey inquired about its objectives, format, curricular time, attendance, faculty development, resources, and evaluation. All directors responded to questions regarding integrating EBM teaching into established educational venues. Of 417 program directors, 269 (65%) responded. Of these 269 programs, 99 (37%) offered a freestanding EBM curriculum. Among these, the most common objectives were performing critical appraisal (78%), searching for evidence (53%), posing a focused question (44%), and applying the evidence in decision making (35%). Although 97% of the programs provided MEDLINE, only 33% provided Best Evidence or the Cochrane Library. Evaluation was performed in 37% of the freestanding curricula. Considering all respondents, most programs reported efforts to integrate EBM teaching into established venues, including attending rounds (84%), resident report (82%), continuity clinic (76%), bedside rounds (68%), and emergency department (35%). However, only 51% to 64% of the programs provided on-site electronic information and 31% to 45% provided site specific faculty development. One third of the training programs reported offering freestanding EBM curricula, which commonly targeted important EBM skills, utilized the residents' experiences, and employed an interactive format. Less than one half of the curricula, however, included curriculum evaluation, and many failed to provide important medical information sources. Most programs reported efforts to integrate EBM teaching, but many of these attempts lacked important structural elements. PMID- 10672118 TI - Racial differences in the utilization of oral anticoagulant therapy in heart failure: a study of elderly hospitalized patients. AB - To assess racial differences in the use of oral anticoagulant therapy for patients with heart failure, we conducted a cohort study of 30 hospitals in northeast Ohio. For 12,911 Medicare enrollees consecutively admitted in 1992 through 1994 with heart failure, crude and adjusted odds of being on oral anticoagulation were determined. The crude and adjusted odds of being African Americans on oral anticoagulant therapy relative to whites were 0.57 (95% confidence interval 0.47-0.69) and 0.55 (95% confidence interval 0. 45-0.67), respectively. African-Americans with heart failure were much less likely than whites to receive oral anticoagulant therapy, even after adjusting for other variables associated with anticoagulant use. PMID- 10672119 TI - Entering the electronic age: risks and challenges for JGIM. PMID- 10672121 TI - Reflections PMID- 10672120 TI - I can't get no patient or practitioner satisfaction. PMID- 10672122 TI - Reflections PMID- 10672123 TI - Journal of internal medicine and the new century. PMID- 10672124 TI - Internal medicine in the 21st century. Introduction. PMID- 10672125 TI - The educated patient: new challenges for the medical profession. AB - The medical profession is facing significant changes in the way the rest of society relates to it. Mass education, mass media and mass consumerism have boomed in the 20th century, putting an increasing amount of pressure on professionals to meet rising public expectations. If doctors are to continue to provide a service that meets the demands of citizens and taxpayers, they need to develop a new relationship with patients, acting not as instructors but as guides, to help people make decisions about their own health. They will have to be more accountable for the quality of care they provide and work with a wider range of health and non-health professionals to meet patients' needs. Doctors need not only to accept the consumer society but also, I will argue, to encourage it. They can work to ensure that the benefits of the information revolution are felt by people excluded from consumerism because of poverty and social isolation, working to create an empowered, informed public whose members are given the best opportunity to look after their own health. PMID- 10672126 TI - Epidemics of vascular toxicity and pulmonary hypertension: what can be learned? AB - Epidemics of vascular disease caused by toxins and infectious agents affecting both humans and animals have been common in history. Examples of agents implicated include anorexients, ergotamine, mercury, arsenic, vinyl chloride, thorotrast, plant alkaloids, nitrites, toxic oil, tryptophan and bacterial, viral and parasitic infections. A major characteristic of these disorders is endothelial dysfunction, which may manifest itself in vasospastic disorders, sclerodermiform skin lesions, fibrosis, osteolytic lesions, polyneuropathy and portal and pulmonary hypertension. Angiosarcoma may also be a late outcome. These diseases are more common than is generally appreciated. The aetiology is usually multifactorial. This and other factors contribute to delayed recognition. PMID- 10672127 TI - Long-term outcome of the Malmo preventive project: mortality and cardiovascular morbidity. AB - OBJECTIVES: To analyse the effects on mortality and cardiovascular morbidity in a population-based sample, invited to an intervention programme incorporating a baseline screening examination and treatment programmes for subjects with cardiovascular risk factors, high alcohol intake and, in women, suspicion of breast cancer on mammography. SETTING: Section of Preventive Medicine, Department of Medicine, University Hospital, Malmo, Sweden. SUBJECTS: Birth cohorts (aged 32 51 years) invited to screening examination (men = 9. 923; women = 4.422) were compared to birth cohorts not invited (men = 6.655; women = 4.290). Mean participation rate in the invited cohorts was 71% (range 64-78%). SCREENING EXAMINATION: Between 1974 and 1992 a baseline screening including a physical examination, blood pressure, a questionnaire regarding, e.g. family history, lifestyle, and socio-economic factors, laboratory tests of serum cholesterol, triglycerides, gamma-glutamyl-transferase, blood glucose before and after an oral glucose load, as well as a mammography examination in women, was performed. INTERVENTIONS: Subjects with hypertension; hyperlipidaemia; diabetes or glucose intolerance; high alcohol intake; or, in women, suspicion of breast cancer were referred to special outpatient clinics. MAIN OUTCOME MEASURES: Total and cause specific mortality, nonfatal myocardial infarction, and stroke, from the screening examination until the end of 1995, was followed in both the intervention and control groups, using national and/or local registries. RESULTS: Total mortality did not differ significantly between the intervention group and control group. Cause-specific deaths were also similar except for 'other' deaths amongst men being significantly lower in the intervention group, mainly due to a lower mortality from 'other' causes (suicide, alcohol related deaths) in men under 40 years of age at baseline. Women under 40 years of age had a significantly lower mortality from cancer in the intervention group than in the control group. Nonfatal myocardial infarction and stroke did not differ between intervention and control group in either sex. Within the invited birth cohorts, nonparticipants had a higher total and cause-specific mortality. CONCLUSIONS: Risk factor screening for major diseases such as cardiovascular disease, alcohol abuse, diabetes mellitus and breast cancer, and subsequent treatment of the detected risk factors/diseases - The Malmo Preventive Project - did not reduce total mortality in the intervention group as a whole. In subjects under 40 years of age at entry, total mortality was lower in the intervention group than in the control group. In men, this seemed to be due to a reduction of alcohol-related deaths, whilst in women death from cancer was reduced. PMID- 10672128 TI - Meal pattern and risk factor evaluation in one-year completers of a weight reduction program for obese men - the 'Gustaf' study. AB - OBJECTIVES: To evaluate changes in meal patterns and in obesity related risk factors after 1 year of treatment in obese men. DESIGN: Data from two 24-h dietary recalls, performed at base-line and after 1 year of treatment, were related to changes in medical risk factors. SETTING: Academic obesity unit. SUBJECTS: Sixty-three men, aged 44 (eight) years (mean [SD]) and Base-line Body Mass Index (BMI) 37.4 (4.6) kg m-2, who had completed 1 year of treatment. The men were subdivided by tertiles according to weight change: tertile I (n = 21), mean +0.3 kg, tertile II (n = 21), mean -5.8 kg and tertile III (n = 21), mean 14.2 kg. MAIN OUTCOME MEASURES: Weight loss, changes in meal patterns and in obesity related medical risk factors. RESULTS: The reported mean energy intake decreased after treatment in tertiles II and III by 700 (1300) kcal (P < 0.05) and 700 (900) kcal (P = 0.001), respectively. In tertile III the energy-% from fat decreased (P < 0.05) with a reciprocal increase in energy-% from protein (P < 0.05). The frequency of snacks of a low nutritional quality decreased (P < 0.01) in tertile III together with an increase in energy-% from 'hot meals of good quality' (P < 0.05). Obesity related risk factors (anthropometry, blood pressure, serum lipid concentrations, blood glucose and plasma insulin) improved in a beneficial way only in tertile III. CONCLUSIONS: The weight loss in the successful tertile III men was to a great extent explained by fewer low quality snacks but more energy from high quality meals. These changes reflected the behaviour modification strategy recommended. PMID- 10672129 TI - Relation between incidence of pneumonia and protective reflexes in post-stroke patients with oral or tube feeding. AB - OBJECTIVES: Although attenuated protective reflexes have been implicated in the development of aspiration pneumonia, the relation between the incidence of pneumonia and the state of these reflexes has not been investigated. Furthermore, the role of feeding tube placement in preventing pneumonia in patients with attenuated protective reflexes is unknown. We studied the relationship between the incidence of pneumonia and the state of cough and swallowing reflexes in post stroke patients with oral or tube feeding. DESIGN: The incidence of pneumonia was prospectively analysed for 1 year in three groups of post-stroke patients on the basis of the following clinical conditions: oral feeding without dysphagia (n = 43); oral feeding with dysphagia (n = 48); and nasogastric tube feeding with dysphagia (n = 52). We also studied the incidence of pneumonia in bedridden patients with nasogastric tube feeding (n = 14). Before the start of the study, the swallowing and cough reflexes of each patient were measured. The swallowing reflex was evaluated according to latency of response, which was timed from the injection of 1 mL of distilled water into the pharynx through a nasal catheter to the onset of swallowing. The cough threshold of citric acid aerosols was defined as the concentration at which the patient coughed five times. RESULTS.;: The incidence of pneumonia was observed in patients having both a latency of response longer than 5 s and a cough threshold for citric acid higher than a concentration of 1.35 (log mg mL-1). The incidence of pneumonia was significantly higher in patients with oral feeding than in those with tube feeding (54.3 vs. 13.2%, P < 0.001). In bedridden patients with tube feeding, the latency of response was longer than 20 s and no patient coughed at the highest concentration of citric acid. The incidence of pneumonia was 64.3% in such patients. CONCLUSIONS: The state of protective reflexes had a significant relation to the incidence of pneumonia. Feeding tube placement may have a beneficial role in preventing aspiration pneumonia in mildly or moderately disabled post-stroke patients with attenuated protective reflexes. Bedridden patients who were tube-fed had the highest incidence of pneumonia. PMID- 10672130 TI - Plasma lipoprotein particle concentrations in postmenopausal women with unstable coronary artery disease. Analysis of diagnostic accuracy using receiver operating characteristics. AB - BACKGROUND: The contribution of plasma lipids to cardiovascular risk is usually evaluated by measuring plasma concentrations of total cholesterol, triglycerides and HDL cholesterol, and calculating LDL cholesterol concentration. We investigated plasma concentrations of apolipoproteins and lipoprotein particles in women with unstable coronary artery disease (CAD) to evaluate whether these, better than the routine lipid status, could differentiate women with and without coronary atherosclerosis. METHODS: Blood samples for lipid analyses were collected from 119 angiographically examined postmenopausal 49-79-year-old women with unstable CAD, and from 101 age-matched controls. Mean plasma concentrations were compared and the discriminatory ability of the different variables were tested using receiver operating characteristics (ROC). RESULTS: At coronary angiography 19% had normal vessels and 81% had coronary atherosclerosis. A disturbed triglyceride metabolism was the most pronounced lipid abnormality in women with unstable CAD and coronary atherosclerosis. ROC showed that none of the evaluated variables had a particularly high discriminatory power regarding unstable CAD or coronary atherosclerosis. The ratio cholesterol/HDL cholesterol was best with an ROC area of 0.79. Furthermore, the newer lipid variables, i.e. lipoprotein particles and apolipoproteins, were no better than the traditional variables. CONCLUSION: Lipoprotein changes reflecting a disturbed triglyceride metabolism are most pronounced in women with unstable CAD and coronary atherosclerosis. Lipoprotein particles and apolipoproteins alone were no better than lipids and lipoproteins in separating women with from those without coronary atherosclerosis. Our study does not support the measurement of apolipoproteins and lipoprotein particles on the basis of diagnostic accuracy alone. PMID- 10672131 TI - Trends in lipid levels and hypercholesterolemia in hypertensive and normotensive finnish adults from 1982 to 1997. AB - OBJECTIVES: To assess the trends in lipid levels and awareness of hypercholesterolemia in hypertensive and normotensive population in Finland from 1982 to 97. DESIGN: Four independent cross-sectional population surveys conducted in 1982, 1987, 1992 and 1997. SETTING: The provinces of North Karelia and Kuopio in eastern Finland and the region of Turku-Loimaa in south-western Finland. SUBJECTS: Men and women aged 25-64 years, selected randomly from the national population register. The subjects were classified to four groups according to their blood pressure level and treatment status: normotensive, unaware hypertensive, aware but not treated hypertensive and treated hypertensive. The total number of participants was 24 083. MAIN OUTCOME MEASURES: We assessed the mean serum total cholesterol and HDL cholesterol concentrations, the prevalence of hypercholesterolemia (total cholesterol >/=6.5 mmol L-1 or the use of lipid lowering drugs), the prevalence of a high ratio of total cholesterol to HDL cholesterol (ratio >/=5) and the awareness of hypercholesterolemia amongst the four study groups. RESULTS: Mean total cholesterol, the prevalence of hypercholesterolemia and the prevalence of a high ratio of total to HDL cholesterol decreased, whereas the awareness of hypercholesterolemia increased significantly in all study groups. The decline in mean total cholesterol was largest in treated hypertensive subjects of both sexes (13% in men, 14% in women). Mean HDL cholesterol increased significantly in all study groups except in the unaware hypertensive men, but it remained significantly lower in treated hypertensive patients in both sexes compared with the other groups (P < 0.001). CONCLUSIONS: The lipid profile of both the hypertensive and normotensive population has significantly improved in both the hypertensive and normotensive population in Finland from 1982-97. The introduction of newer antihypertensive drugs has not had any effect on the HDL cholesterol level amongst treated hypertensive patients at the population level. PMID- 10672132 TI - Altered bone metabolism in inflammatory bowel disease: there is a difference between Crohn's disease and ulcerative colitis. AB - OBJECTIVES: The aims of this study were to assess bone metabolism in inflammatory bowel disease (IBD) patients and to evaluate potential differences between Crohn's disease (CD) and ulcerative colitis (UC) with respect to the mechanisms underlying bone loss in this group of diseases. DESIGN AND SETTING: This was a cross-sectional study which started in 1992. Patients were randomly selected for invitation to participate and were examined during the years 1992-95 in one research clinic in Milan. SUBJECTS AND METHODS: Fifty-one patients suffering from CD (30 women and 21 men, mean age 38.7 +/- 13.2 years) and 40 with UC (15 women and 25 men, mean age 34.4. +/- 12.5 years) entered the study. Thirty healthy subjects were selected as sex- and age-matched controls (C). Spine and femoral neck bone mineral density (expressed as T score), calciotropic hormones (parathyroid hormone, PTH; 25-hydroxycholecalciferol, 25(OH)D3; 1,25 hydroxycholecalciferol, 1, 25(OH)D3) and biochemical markers of bone turnover (ostecalcin, OC; total alkaline phosphatase, ALP; type I collagen C-terminal telopeptide, ICTP) were evaluated. RESULTS: Spine and femur T scores were similar in the two groups (spine: CD = -1.49 +/- 1.46; UC = -1. 67 +/- 1.13; femur: CD = 1.80 +/- 1.36; UC = -1.60 +/- 1.03). Based upon the WHO guidelines, only 8% of CD patients and 15% of UC patients had a normal bone mineral density (BMD), 55% (CD) and 67% (UC) were osteopenic, and 37% (CD) and 18% (UC) were osteoporotic. The distribution amongst the three different diagnostic groups was not significantly different between CD and UC groups (P = 0.11). PTH and 25(OH)D3 concentrations were not significantly different between CD and UC patients and controls, whilst 1,25(OH)D3 concentrations were significantly lower in both CD and UC patients compared with controls (P < 0.05). Bone turnover was increased in UC but not in CD patients, as shown by significantly increased concentrations in UC patients of both OC (CD = 7.77 +/- 5.06, UC = 10.03 +/- 6.24, C = 6. 58 +/- 2.87, P < 0.05 vs. C) and ICTP (CD = 5.74 +/- 3.94, UC = 10.2 +/- 8.47, C = 3.48 +/- 0.95, P < 0.05 vs. CD and C). In a stepwise regression that included age, sex, disease duration and cumulative prednisolone dose as independent variables, the femur T score was significantly inversely related to disease duration (r2 = 0.125, F = 6.06) in CD patients. In UC patients, the spine T score was inversely related to age (r2 = 0.107, F = 5.49) and significantly related to sex (more negative in males: r2 = 0.3, F = 16.1); the femur T score was significantly related to sex (more negative in males) and inversely related to the cumulative prednisolone dose (r2 = 0.283, F = 7.3). CONCLUSIONS: These data show that IBD patients have a diffuse osteopenia, the degree of which is not different in CD and UC; however, bone turnover is significantly higher in UC. Finally, osteopenia is related to disease duration in CD, whilst it is related to the male sex and glucocorticoid treatment in UC. PMID- 10672133 TI - Angiotensin converting enzyme (ACE) gene polymorphism in sarcoidosis in relation to associated autoimmune diseases. AB - OBJECTIVES: To investigate the significance of ACE gene insertion/deletion (I/D) polymorphism in the frequency of autoimmune manifestations in sarcoidosis. DESIGN: In patients with sarcoidosis the ACE gene I/D polymorphism was detected with PCR on genomic DNA. The patients with sarcoidosis were divided according to the presence (n = 30) or absence (n = 32) of autoimmune manifestations. The former group was subdivided into thyroid autoimmunity (n = 10), gluten immune reactivity (n = 10) and gastric autoimmunity (n = 17). SETTINGS: The patients were recruited at the Department of Pulmonary Medicine, and the study was conducted at the Department of Endocrinology, University of Lund, Malmo University Hospital, Malmo, Sweden. SUBJECTS: Sixty-two patients with documented sarcoidosis (30 females, 32 males, median age/range at diagnosis of sarcoidosis 31.5/19-75 years, median age/range at study 47.5/22-81 years) were examined. A total of 107 healthy unrelated subjects without sarcoidosis (60 females, 47 males, median age/range at study 58/40-82 years) served as controls. RESULTS: S ACE values were significantly increased in patients compared to controls (P = 0.00001). The same was true in the subgroup of sarcoidosis patients with associated autoimmunity compared with those with isolated sarcoidosis (P = 0.0328). A significant association was seen between ACE gene polymorphism (II, ID, DD genotypes) and S-ACE levels in both patients and controls according to the order II < ID < DD. The observed genotype frequency distributions in the different study groups agreed the Hardy-Weinberg equilibrium without significant differences between the patients and the controls. Within the group with autoimmune manifestations the DD genotype was significantly over-represented in X ray stage III compared to the other X-ray stages (P = 0.0181) and a significant increase in the DD genotype in X-ray stage III (P = 0.035) in the group with autoimmune manifestations compared to isolated sarcoidosis was detected. CONCLUSION: We confirmed that the S-ACE levels corresponded to the order II < ID < DD in patients with sarcoidosis as well as in healthy controls. S-ACE levels were significantly higher in sarcoidosis patients with autoimmune manifestations. The frequency of the DD genotype was significantly increased in patients with autoimmune manifestations and major granuloma mass (X-ray stage III). The ACE D allele in its homozygous form may confer susceptibility for autoimmune manifestations in sarcoidosis, possibly via the high levels of S-ACE it encodes. PMID- 10672134 TI - Hypomagnesemia in heart failure with ventricular arrhythmias. Beneficial effects of magnesium supplementation. AB - OBJECTIVE: To assess the role of electrolyte imbalance in cardiac arrhythmias associated with congestive heart failure. DESIGN: Serum magnesium and potassium levels, urine magnesium excretion and the incidence of ventricular arrhythmias were assessed throughout the study. The patients who displayed complex arrhythmias after the first week of hospital medication were randomized 2:1 to double-blind magnesium supplementation or placebo. SETTING: The study was carried out in one municipal hospital, providing primary care. SUBJECTS: A total of 588 consecutive patients were screened for eligibility (clinical heart failure >/=6 months; NYHA class II-IV; left ventricular ejection fraction /=15 mmHg after intervention. RESULTS: A DBP response was seen in 24 patients. In 11 patients, invasive treatment did not result in a DBP response or failed technically. Compared with these patients, the responders were younger (55 +/- 11 vs. 66 +/- 8 years, P = 0.001) and tended to have higher DBP (100 +/- 8 vs. 93 +/- 11 mmHg, P = 0.065). The function of the affected kidney, or that of the more affected kidney if RAD was bilateral, was better preserved in responders (relative clearance on captopril renography 23 +/- 15 vs. 8 +/- 4%, P = 0.008). A response was more often seen in unilateral than in bilateral RAD (81% vs. 33%, P = 0.015). A relative clearance of 6% uncorrected sequence divergence) was found to exist between populations found southeast of the Colorado River (eastern Arizona, Colorado, Wyoming and New Mexico) and all other western populations. Other well-supported subclades were composed of samples from: (i) the islands and north coast of southeast Alaska; (ii) eastern Alaska, British Columbia, Washington and Oregon; and (iii) northern California, Idaho and Montana. Within subclades, divergence was low. Our results suggest that the close relationships among haplotypes within northern subclades are a result of recent colonization, whereas higher among-subclade divergence is caused by genetic differentiation during prolonged periods of isolation, possibly as a result of mid-Pleistocene climatic events. PMID- 10672161 TI - Species-specific oligonucleotide probes for macroalgae: molecular discrimination of two marine fouling species of Enteromorpha (Ulvophyceae). AB - The green seaweeds Enteromorpha intestinalis and E. compressa are important fouling organisms commonly found in polluted and nutrient-enriched marine and brackish water habitats, where they are used in environmental monitoring. Discrimination of the two species is extremely difficult because of overlapping morphological characters. In this study a quick molecular method for species identification was developed based on the nuclear rDNA ITS2 sequence data of 54 E. intestinalis samples and 20 E. compressa samples from a wide geographical range. Oligonucleotide probes were designed for species-specific hybridization to dot-blots of the PCR-amplified ITS1, 5.8S gene and ITS2 fragment of both E. intestinalis and E. compressa. Specificity of the oligonucleotide probes was confirmed by tests with taxonomically diverse species that could morphologically be confused with E. intestinalis or E. compressa. This is the first use of species-specific probes for macroalgae. The restriction endonuclease NruI digested specifically the amplified PCR product from E. compressa into two fragments detectable on agarose gels, but no suitable restriction sites were identifiable in the PCR product of E. intestinalis. PMID- 10672162 TI - Sex ratio and fledging success of supplementary-fed Tengmalm's owl broods. AB - A nest box population of Tengmalm's owls (Aegolius funereus) in northern Sweden was studied to investigate the effects of extra food on the sex ratio between hatching and fledging in this sexually size-dimorphic species. The brood size and brood sex ratio of supplementary-fed and control broods were compared. Newly hatched nestlings were blood sampled and sexed by polymerase chain reaction (PCR) amplification of the sex-linked CHD1Z and CHD1W genes. The brood sex ratio at hatching was strongly male biased (65%); this was also the case in broods where all eggs hatched (72%). There was no relationship between hatch order and sex ratio, and hatching sex ratio did not vary significantly with laying date. Brood size decreased between hatching and fledging, but did not differ between fed and control broods at either stage. Brood sex ratio did not differ between hatching and fledging, and fledging sex ratio did not differ between fed and control broods. It was concluded that, at least during the year in which the study was carried out, feeding had no effect on brood reduction, and that male and female nestlings did not show any differential mortality. The mechanisms behind the male biased sex ratio at hatching, and any possible adaptive reasons for it, are not known. PMID- 10672163 TI - A phylogenetic analysis of body size evolution in the Anolis roquet group (Sauria: Iguanidae): character displacement or size assortment? AB - The important role that competition plays in structuring communities is well documented; however, the role of competition in an evolutionary context remains unclear. Evolutionary investigations into the role of competition have often focused on the process of character displacement, and a good example of this is the evolution of body size in the Anolis lizards of the Caribbean islands. Previous work on the A. roquet species group has taken a phylogenetic approach and concluded that patterns of body size differences are not caused by character displacement but are a result of size assortment. Using a phylogenetic reconstruction based on the sequence of the cytochrome b gene (cyt-b) and ancestral character-state reconstruction methods, we investigated the roles of character displacement and size assortment. Our results indicated that size assortment alone was insufficient to explain the observed patterns of body size differences. Furthermore, we found that change in body size was associated with a change in allopatry/sympatry, thus supporting the character-displacement hypothesis. We conclude that patterns of body size differences in the A. roquet species group appear to be the result of a combination of character displacement and size assortment because character displacement was only found to be possible on three occasions. PMID- 10672164 TI - Sex, parthenogenesis and genetic structure of rotifers: microsatellite analysis of contemporary and resting egg bank populations. AB - Cyclically parthenogenetic rotifers are a valuable model for investigating the relationship between reproductive mode and population structure, although advances in this field have been hindered by low allozyme variability in these organisms. A high genotypic diversity is predicted after population establishment, which would be eroded by clonal selection during the parthenogenetic phase. The resting egg bank, produced sexually, is presumed to store high levels of genetic diversity, with subsequent effects on planktonic population structure. Here, we provide the first application of microsatellite markers to a rotifer planktonic population and its associated resting egg bank. Seven polymorphic microsatellite loci were screened in populations of the rotifer Brachionus plicatilis in a temporary pond to analyse: (i) the genetic structure of the resting egg bank; (ii) the changes in the genetic structure of rotifer populations during the parthenogenetic phase; and (iii) the population structure after its initiation from resting eggs. Microsatellites proved to be a useful tool for clone identification, revealing a surprisingly high clonal diversity in rotifer populations. The last sample in the parthenogenetic phase showed evidence of clonal selection, as indicated by a low observed clonal diversity and the appearance of linkage disequilibria. The resting egg bank, analysed comprehensively for the first time in any zooplankter, is in Hardy-Weinberg and linkage equilibrium, and contains a high genotypic diversity. Unexpectedly, the resting egg bank differed from the planktonic population in its allelic composition, suggesting that resting egg hatching is biased. PMID- 10672165 TI - Screening of Mhc variation in Atlantic salmon (Salmo salar): a comparison of restriction fragment length polymorphism (RFLP), denaturing gradient gel electrophoresis (DGGE) and sequencing. AB - We compared three different molecular methods currently used for screening of Mhc variation in population studies of Atlantic salmon. Restriction fragment length polymorphism (RFLP) of the entire class II gene detected 22 haplotypes. Seventeen exon 2 sequences were obtained from individuals carrying the 22 haplotypes, two of which had not been detected by RFLP. The six alleles (27%) detected by RFLP and not by exon 2 sequencing probably resulted from sequence variation outside exon 2. Within exon 2, RFLP differentiated 88% of the sequences. Alternatively, denaturing gradient gel electrophoresis (DGGE) performed under two run conditions detected 94% of the sequence variation. Both RFLP using different probes, and the two PCR-based methods using three different primer pairs, suggest that there is only a single Mhc class II B gene in the Baltic populations of Atlantic salmon. PMID- 10672166 TI - Genetic divergence between Atlantic and Indo-Pacific stocks of bigeye tuna (Thunnus obesus) and admixture around South Africa. AB - Two mitochondrial DNA segments of the bigeye tuna (Thunnus obesus) were amplified by polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLP) analyses of these segments were used for the genetic stock study. The variation in a segment flanking the ATPase and COIII genes was low; only two genotypes (alpha and beta) were detected by RsaI digestion. Yet a large difference in the genotype distribution was observed between ocean basin samples. The alpha type predominated in four Atlantic samples, where 178 of 244 individuals were the alpha type. In contrast, only one of 195 individuals collected in the Indo-Pacific was the alpha type? The frequency of the alpha type varied considerably from 0 to 80% among seven samples collected off the Cape of Good Hope. The variation found in the other segment, containing the D-loop region, was much higher; two endonucleases (DpnII and RsaI) detected five genotypes each and 15 composite genotypes. A highly significant difference in genotype frequencies was observed between the Atlantic and Indo-Pacific samples, but no heterogeneity was observed among the four Atlantic or among four Indo Pacific samples. These results clearly indicate that not only gene flow, but also fish migration, between the Atlantic and Indian Oceans are severely restricted, and that fishes from these distinct stocks are intermingling around South Africa. The simple and diagnostic genetic marker found in this study can be used to estimate mixing ratios between Atlantic and Indian stocks around South Africa. PMID- 10672167 TI - The use of random amplified polymorphic DNA (RAPD) markers to identify strawberry varieties: a forensic application. AB - The random amplified polymorphic DNA (RAPD) technique was applied to settle a lawsuit involving unauthorized commercialization of a patented strawberry variety of high economical relevance ('Marmolada'). Because of economical involvements, the molecular approach was added to the more traditional morphological examination in a double-blind test. All plants belonging to the patented variety were unambiguously identified (13 plants among a total of 31 plants examined). The results were accepted as evidence in the court. This study confirms that the RAPD technique is especially suitable for identification of asexually reproduced plant varieties for forensic or agricultural purposes. PMID- 10672168 TI - Screening for intron-length polymorphisms in penaeid shrimps using exon-primed intron-crossing (EPIC)-PCR. PMID- 10672169 TI - Polymorphic microsatellites in the common cuttlefish Sepia officinalis (Cephalopoda). PMID- 10672170 TI - Development of tri- and tetranucleotide repeat microsatellite loci in Atlantic cod (Gadus morhua). PMID- 10672171 TI - Characterization of microsatellite loci in Sebastes alutus and their conservation in congeneric rockfish species. PMID- 10672172 TI - Highly polymorphic microsatellite markers in the white-lined bat (Saccopteryx bilineata). PMID- 10672173 TI - beta-1,6-Glucan synthesis in Saccharomyces cerevisiae. AB - beta-1,6-Glucan is an essential fungal-specific component of the Saccharomyces cerevisiae cell wall that interconnects all other wall components into a lattice. Considerable biochemical and genetic effort has been directed at the identification and characterization of the steps involved in its biosynthesis. Structural studies show that the polymer plays a central role in wall structure, attaching mannoproteins via their glycosylphosphatidylinositol (GPI) glycan remnant to beta-1,3-glucan and chitin. Genetic approaches have identified genes that upon disruption result in beta-1,6-glucan defects of varying severity, often with reduced growth or lethality. These gene products have been localized throughout the secretory pathway and at the cell surface, suggesting a possible biosynthetic route. Current structural and genetic data have therefore allowed the development of models to predict biosynthetic events. Based on knowledge of beta-1,3-glucan and chitin synthesis, it is likely that the bulk of beta-1,6 glucan polymer synthesis occurs at the cell surface, but requires key prior intracellular events. However, the activity of most of the identified gene products remain unknown, making it unclear to what extent and how directly they contribute to the synthesis of this polymer. With the recent availability of new tools, reagents and methods (including genomics), the field is poised for a convergence of biochemical and genetic methods to identify and characterize the biochemical steps in the synthesis of this polymer. PMID- 10672174 TI - A bacterial genome in flux: the twelve linear and nine circular extrachromosomal DNAs in an infectious isolate of the Lyme disease spirochete Borrelia burgdorferi. AB - We have determined that Borrelia burgdorferi strain B31 MI carries 21 extrachromosomal DNA elements, the largest number known for any bacterium. Among these are 12 linear and nine circular plasmids, whose sequences total 610 694 bp. We report here the nucleotide sequence of three linear and seven circular plasmids (comprising 290 546 bp) in this infectious isolate. This completes the genome sequencing project for this organism; its genome size is 1 521 419 bp (plus about 2000 bp of undetermined telomeric sequences). Analysis of the sequence implies that there has been extensive and sometimes rather recent DNA rearrangement among a number of the linear plasmids. Many of these events appear to have been mediated by recombinational processes that formed duplications. These many regions of similarity are reflected in the fact that most plasmid genes are members of one of the genome's 161 paralogous gene families; 107 of these gene families, which vary in size from two to 41 members, contain at least one plasmid gene. These rearrangements appear to have contributed to a surprisingly large number of apparently non-functional pseudogenes, a very unusual feature for a prokaryotic genome. The presence of these damaged genes suggests that some of the plasmids may be in a period of rapid evolution. The sequence predicts 535 plasmid genes >/=300 bp in length that may be intact and 167 apparently mutationally damaged and/or unexpressed genes (pseudogenes). The large majority, over 90%, of genes on these plasmids have no convincing similarity to genes outside Borrelia, suggesting that they perform specialized functions. PMID- 10672175 TI - Acid- and multistress-resistant mutants of Lactococcus lactis : identification of intracellular stress signals. AB - Lactococcus lactis growth is accompanied by lactic acid production, which results in acidification of the medium and arrest of cell multiplication. Despite growth limitation at low pH, there is evidence that lactococci do have inducible responses to an acid pH. In order to characterize the genes involved in acid tolerance responses, we selected acid-resistant insertional mutants of the L. lactis strain MG1363. Twenty-one independent characterized mutants were affected in 18 different loci, some of which are implicated in transport systems or base metabolism. None of these genes was identified previously as involved in lactococcal acid tolerance. The various phenotypes obtained by acid stress selection allowed us to define four classes of mutants, two of which comprise multistress-resistant strains. Our results reveal that L. lactis has several means of protecting itself against low pH, at least one of which results in multiple stress resistance. In particular, intracellular phosphate and guanine nucleotide pools, notably (p)ppGpp, are likely to act as signals that determine the level of lactococcal stress response induction. Our results provide a link between the physiological state of the cell and the level of stress tolerance and establish a role for the stringent response in acid stress response regulation. PMID- 10672176 TI - Genetic and biochemical evidence of a Campylobacter jejuni capsular polysaccharide that accounts for Penner serotype specificity. AB - Campylobacter jejuni, a Gram-negative spiral bacterium, is the most common bacterial cause of acute human gastroenteritis and is increasingly recognized for its association with the serious post-infection neurological complications of the Miller-Fisher and Guillain-Barre syndromes. C. jejuni lipopolysaccharide (LPS) is thought to be involved in the pathogenesis of both uncomplicated infection and more serious sequelae, yet the LPS remains poorly characterized. Current studies on C. jejuni suggest that all strains produce lipooligosaccharide (LOS), with about one-third of strains also producing high-molecular-weight LPS (referred to as O-antigen). In this report, we demonstrate the presence of the high-molecular weight LPS in all C. jejuni strains tested. Furthermore, we show that this LPS is biochemically and genetically unrelated to LOS and is similar to group II and group III capsular polysaccharides. All tested kpsM, kpsS and kpsC mutants of C. jejuni lost the ability to produce O-antigen. Moreover, this correlated with serotype changes. We demonstrate for the first time that the previously described O-antigen of C. jejuni is a capsular polysaccharide and a common component of the thermostable antigen used for serotyping of C. jejuni. PMID- 10672177 TI - Trypanosomes lacking trypanothione reductase are avirulent and show increased sensitivity to oxidative stress. AB - In Kinetoplastida, trypanothione and trypanothione reductase (TRYR) provide an intracellular reducing environment, substituting for the glutathione-glutathione reductase system found in most other organisms. To investigate the physiological role of TRYR in Trypanosoma brucei, we generated cells containing just one trypanothione reductase gene, TRYR, which was under the control of a tetracycline inducible promoter. This enabled us to regulate TRYR activity in the cells from less than 1% to 400% of wild-type levels by adjusting the concentration of added tetracycline. In normal growth medium (which contains reducing agents), trypanosomes containing less than 10% of wild-type enzyme activity were unable to grow, although the levels of reduced trypanothione and total thiols remained constant. In media lacking reducing agents, hypersensitivity towards hydrogen peroxide (EC50 = 3.5 microM) was observed compared with the wild type (EC50 = 223 microM). The depletion of TRYR had no effect on susceptibility to melarsen oxide. The infectivity and virulence of the parasites in mice was dependent upon tetracycline-regulated TRYR activity: if the trypanosomes were injected into mice in the absence of tetracycline, no infection was detectable; and when tetracycline was withdrawn from previously infected animals, the parasitaemia was suppressed. PMID- 10672178 TI - Differential post-transcriptional regulation of yeast mRNAs in response to high and low glucose concentrations. AB - Glucose regulates yeast gene expression at both transcriptional and post transcriptional levels. Glucose strongly represses the transcription of the gluconeogenic genes, FBP1 and PCK1, and accelerates the degradation of their mRNAs. Together these mechanisms are responsible for the rapid decrease in gluconeogenic enzyme synthesis when yeast cells switch to glycolytic metabolism. In this study, we show that accelerated gluconeogenic mRNA degradation can be triggered by low concentrations of glucose (<0. 02%). This sets the FBP1 and PCK1 mRNAs apart from other glucose-sensitive mRNAs, such as the Ip mRNA, which only responds to high glucose concentrations (>1%). We also show that accelerated gluconeogenic mRNA degradation is co-ordinated with transcriptional repression by common signalling components that include sugar kinases and Ras-cAMP signalling. Furthermore, the ability of the low glucose signal to trigger accelerated gluconeogenic mRNA degradation depends upon the low glucose sensor, Snf3p, but not on the high glucose sensor, Rgt2p. Also, this response is influenced by reg1 and ume5 mutations, but not by grr1 or rgt1 mutations. Our data suggest that several signalling pathways co-ordinate differential post-transcriptional and transcriptional responses in yeast, depending upon the amount of glucose available in the medium. PMID- 10672179 TI - A genomic analysis of two-component signal transduction in Streptococcus pneumoniae. AB - A genomics-based approach was used to identify the entire gene complement of putative two-component signal transduction systems (TCSTSs) in Streptococcus pneumoniae. A total of 14 open reading frames (ORFs) were identified as putative response regulators, 13 of which were adjacent to genes encoding probable histidine kinases. Both the histidine kinase and response regulator proteins were categorized into subfamilies on the basis of phylogeny. Through a systematic programme of mutagenesis, the importance of each novel TCSTS was determined with respect to viability and pathogenicity. One TCSTS was identified that was essential for the growth of S. pneumoniaeThis locus was highly homologous to the yycFG gene pair encoding the essential response regulator/histidine kinase proteins identified in Bacillus subtilis and Staphylococcus aureus. Separate deletions of eight other loci led in each case to a dramatic attenuation of growth in a mouse respiratory tract infection model, suggesting that these signal transduction systems are important for the in vivo adaptation and pathogenesis of S. pneumoniae. The identification of conserved TCSTSs important for both pathogenicity and viability in a Gram-positive pathogen highlights the potential of two-component signal transduction as a multicomponent target for antibacterial drug discovery. PMID- 10672180 TI - The Brucella abortus Lon functions as a generalized stress response protease and is required for wild-type virulence in BALB/c mice. AB - The gene encoding a Lon protease homologue has been cloned from Brucella abortus. The putative Brucella abortus Lon shares > 60% amino acid identity with its Escherichia coli counterpart and the recombinant form of this protein restores the capacity of an Escherichia coli lon mutant to resist killing by ultraviolet irradiation and regulate the expression of a cpsB:lacZ fusion to wild-type levels. A sigma32 type promoter was identified upstream of the predicted lon coding region and Northern analysis revealed that transcription of the native Brucella abortus lon increases in response to heat shock and other environmental stresses. ATP-dependent proteolytic activity was also demonstrated for purified recombinant Lon. To evaluate the capacity of the Brucella abortus Lon homologue to function as a stress response protease, the majority of the lon coding region was removed from virulent strain Brucella abortus 2308 via allelic exchange. In contrast to the parent strain, the Brucella abortus lon mutant, designated GR106, was impaired in its capacity to form isolated colonies on solid medium at 41 degrees C and displayed an increased sensitivity to killing by puromycin and H2O2. GR106 also displayed reduced survival in cultured murine macrophages and significant attenuation in BALB/c mice at 1 week post infection compared with the virulent parental strain. Beginning at 2 weeks and continuing for 6 weeks post infection, however, GR106 and 2308 displayed equivalent spleen and liver colonization levels in mice. These findings suggest that the Brucella abortus Lon homologue functions as a stress response protease that is required for wild-type virulence during the initial stages of infection in the mouse model, but is not essential for the establishment and maintenance of chronic infection in this host. PMID- 10672181 TI - A folding variant of alpha-lactalbumin with bactericidal activity against Streptococcus pneumoniae. AB - This study describes an alpha-lactalbumin folding variant from human milk with bactericidal activity against antibiotic-resistant and -susceptible strains of Streptococcus pneumoniae. The active complex precipitated with the casein fraction at pH 4.6 and was purified from casein by a combination of anion exchange and gel chromatography. Unlike other casein components, the active complex was retained on the ion-exchange matrix and eluted only with high salt. The eluted fraction showed N-terminal and mass spectrometric identity with human milk alpha-lactalbumin, but native alpha-lactalbumin had no bactericidal effect. Spectroscopic analysis demonstrated that the active form of the molecule was in a different folding state, with secondary structure identical to alpha-lactalbumin from human milk whey, but fluctuating tertiary structure. Native alpha lactalbumin could be converted to the active bactericidal form by ion-exchange chromatography in the presence of a cofactor from human milk casein, characterized as a C18:1 fatty acid. Analysis of the antibacterial spectrum showed selectivity for streptococci; Gram-negative and other Gram-positive bacteria were resistant. The folding variant of alpha-lactalbumin is a new example of naturally occurring molecules with antimicrobial activity. PMID- 10672182 TI - The cell wall architecture of Candida albicans wild-type cells and cell wall defective mutants. AB - In Candida albicans wild-type cells, the beta1, 6-glucanase-extractable glycosylphosphatidylinositol (GPI)-dependent cell wall proteins (CWPs) account for about 88% of all covalently linked CWPs. Approximately 90% of these GPI-CWPs, including Als1p and Als3p, are attached via beta1,6-glucan to beta1,3-glucan. The remaining GPI-CWPs are linked through beta1,6-glucan to chitin. The beta1,6 glucanase-resistant protein fraction is small and consists of Pir-related CWPs, which are attached to beta1,3-glucan through an alkali-labile linkage. Immunogold labelling and Western analysis, using an antiserum directed against Saccharomyces cerevisiae Pir2p/Hsp150, point to the localization of at least two differentially expressed Pir2 homologues in the cell wall of C. albicans. In mnn9Delta and pmt1Delta mutant strains, which are defective in N- and O-glycosylation of proteins respectively, we observed enhanced chitin levels together with an increased coupling of GPI-CWPs through beta1,6-glucan to chitin. In these cells, the level of Pir-CWPs was slightly upregulated. A slightly increased incorporation of Pir proteins was also observed in a beta1, 6-glucan-deficient hemizygous kre6Delta mutant. Taken together, these observations show that C. albicans follows the same basic rules as S. cerevisiae in constructing a cell wall and indicate that a cell wall salvage mechanism is activated when Candida cells are confronted with cell wall weakening. PMID- 10672183 TI - The putative DNA translocase SpoIIIE is required for sporulation of the symmetrically dividing coccal species Sporosarcina ureae. AB - The spoIIIE gene of Sporosarcina ureae encodes a 780-residue protein, showing 58% identity to the SpoIIIE protein of Bacillus subtilis, which is thought to be a DNA translocase. Expression of the S. ureae spoIIIE gene is able to restore sporulation in a B. subtilis spoIIIE mutant. Inactivation of the S. ureae spoIIIE gene blocks sporulation of S. ureae at stage III. Within the limits of detection, the sporulation division in S. ureae shows the same symmetry, or near symmetry, as the vegetative division (in contrast to the highly asymmetric location of the sporulation division for B. subtilis), and so it is inferred that SpoIIIE facilitates chromosome partitioning during sporulation, even when the division is not grossly asymmetric. It is suggested that chromosome partitioning lags behind division during sporulation but not during vegetative growth. PMID- 10672184 TI - Promoter discrimination by the related transcriptional activators MarA and SoxS: differential regulation by differential binding. AB - MarA and SoxS are closely related proteins ( approximately 45% identical) that transcriptionally activate a common set of unlinked genes, resulting in multiple antibiotic and superoxide resistance in Escherichia coli. Both proteins bind as monomers to a 20 bp degenerate asymmetric recognition sequence, the 'marbox', located upstream of the promoter. However, the proteins differ widely in the extents to which they activate particular promoters, with the consequence that overexpression of SoxS leads to greater superoxide resistance than does overexpression of MarA. This 'discrimination' between activators by promoters was demonstrated in vivo, using promoters fused to lacZ, and in vitro, using purified RNA polymerase, promoter DNA and MarA or SoxS. The marbox was found to be a critical element in discrimination by in vivo and in vitro assays of hybrid promoters containing the marbox from one gene and the core promoter from another. Furthermore, by sequential mutation of its marbox, a promoter that discriminated 35-fold in favour of SoxS was converted into one that did not discriminate. The relative activation of a promoter by MarA or SoxS was paralleled by the relative binding of the two activators to the promoter's marbox as assayed by band shift experiments. Thus, differential recognition of closely related marbox sequences by the closely related activators is the primary basis for promoter discrimination. Discrimination enables the cell to customize its response to the stresses that trigger synthesis of the activators. PMID- 10672186 TI - BasT, a membrane-bound transducer protein for amino acid detection in Halobacterium salinarum. AB - Halophilic archaea, such as eubacteria, use methyl-accepting chemotaxis proteins (MCPs) to sense their environment. We show here that BasT is a halobacterial transducer protein (Htp) responsible for chemotaxis towards five attractant amino acids. The C-terminus of the protein exhibits the highly conserved regions that are diagnostic for MCPs: the signalling domain for communication with the histidine kinase and the methylation sites that interact with the methylation/demethylation enzymes for adaptation. Hydropathy analysis predicts an enterobacterial-type transducer protein topology for BasT, with an extracellular putative ligand-binding domain flanked by two transmembrane helices and a cytoplasmic domain. BasT-inactivated mutant cells are missing a membrane protein radiolabelled with L-[methyl-3H]-methionine in wild-type cells, confirming that BasT is methylatable and membrane bound. Behavioural analysis of the basT mutant cells by capillary and chemical-in-plug assays demonstrates complete loss of chemotactic responses towards five (leucine, isoleucine, valine, methionine and cysteine) of the six attractant amino acids for Halobacterium salinarum, whereas they still respond to arginine. The volatile methyl group production assays also corroborate these findings and confirm that BasT signalling induces methyl group turnover. Our data identify BasT as the chemotaxis transducer protein for the branched chain amino acids leucine, isoleucine and valine as well as for methionine and cysteine. Thus, BasT and the arginine sensor Car cover the entire spectrum of chemotactic responses towards attractant amino acids in H. salinarum. PMID- 10672185 TI - Characterization of two novel regulatory genes affecting Salmonella invasion gene expression. AB - A Salmonella typhimurium chromosomal deletion removing approximately 19 kb of DNA at centisome 65 reduces invasion of cultured epithelial cells as well as the expression of lacZY operon fusions to several genes required for the invasive phenotype. As the deleted region contains no genes previously known to affect Salmonella invasion, we investigated the roles of individual genes in the deleted region using a combination of cloning, complementation and directed mutation. We find that the deletion includes two unrelated regulatory genes. One is the Salmonella homologue of Escherichia coli barA (airS ), which encodes a member of the multistep phosphorelay subgroup of two-component sensor kinases. The action of BarA is coupled to that of SirA, a member of the phosphorylated response regulator family of proteins, and includes both HilA-dependent and HilA independent components. The other regulatory gene removed by the deletion is the Salmonella homologue of E. coli csrB, which specifies a regulatory RNA implicated in controlling specific message turnover in E. coli. These results identify a protein that is likely to play a key role in the environmental control of Salmonella invasion gene expression, and they also suggest that transcriptional control of invasion genes could be subject to refinement at the level of message turnover. PMID- 10672187 TI - The response regulator RssB, a recognition factor for sigmaS proteolysis in Escherichia coli, can act like an anti-sigmaS factor. AB - sigmaS (RpoS) is the master regulator of the general stress response in Escherichia coli. Several stresses increase cellular sigmaS levels by inhibiting proteolysis of sigmaS, which under non-stress conditions is a highly unstable protein. For this ClpXP-dependent degradation, the response regulator RssB acts as a recognition factor, with RssB affinity for sigmaS being modulated by phosphorylation. Here, we demonstrate that RssB can also act like an anti-sigma factor for sigmaS in vivo, i.e. RssB can inhibit the expression of sigmaS dependent genes in the presence of high sigmaS levels. This becomes apparent when (i) the cellular RssB/sigmaS ratio is at least somewhat elevated and (ii) proteolysis is reduced (for example in stationary phase) or eliminated (for example in a clpP mutant). Two modes of inhibition of sigmaS by RssB can be distinguished. The 'catalytic' mode is observed in stationary phase cells with a substoichiometric RssB/sigmaS ratio, requires ClpP and therefore probably corresponds to sequestering of sigmaS to Clp protease (even though sigmaS is not degraded). The 'stoichiometric' mode occurs in clpP mutant cells upon overproduction of RssB to levels that are equal to those of sigmaS, and therefore probably involves binary complex formation between RssB and sigmaS. We also show that, under standard laboratory conditions, the cellular level of RssB is more than 20-fold lower than that of sigmaS and is not significantly controlled by stresses that upregulate sigmaS. We therefore propose that antisigma factor activity of RssB may play a role under not yet identified growth conditions (which may result in RssB induction), or that RssB is a former antisigma factor that during evolution was recruited to serve as a recognition factor for proteolysis. PMID- 10672188 TI - Homologous gene knockout in the archaeon Halobacterium salinarum with ura3 as a counterselectable marker. AB - To facilitate the functional genomic analysis of an archaeon, we have developed a homologous gene replacement strategy for Halobacterium salinarum based on ura3, which encodes the pyrimidine biosynthetic enzyme orotidine-5'-monophosphate decarboxylase. H. salinarum was shown to be sensitive to 5-fluoroorotic acid (5 FOA), which can select for mutations in ura3. A spontaneous 5-FOA-resistant mutant was found to contain an insertion in ura3 and was a uracil auxotroph. Integration of ura3 at the bacterioopsin locus (bop ) of this mutant restored 5 FOA sensitivity and uracil prototrophy. Parallel results were obtained with a Deltaura3 strain constructed by gene replacement and with derivatives of this strain in which ura3 replaced bop. These results show that H. salinarum ura3 encodes functional orotidine-5'-monophosphate decarboxylase. To demonstrate ura3 based gene replacement, a Deltabop strain was constructed by transforming a Deltaura3 host with a bop deletion plasmid containing a mevinolin resistance marker. In one approach, the host contained intact ura3 at the chromosomal bop locus; in another, ura3 was included in the plasmid. Plasmid integrants selected with mevinolin were resolved with 5-FOA, yielding Deltabop recombinants at a frequency of > 10-2 in both approaches. These studies establish an efficient new genetic strategy towards the systematic knockout of genes in an archaeon. PMID- 10672189 TI - A chromosomally encoded regulator is required for expression of the Yersinia enterocolitica inv gene and for virulence. AB - The primary invasion factor of Yersinia enterocolitica, invasin, is encoded by inv. inv expression is regulated in response to pH, growth phase and temperature. In vitro, inv is maximally expressed at 26 degrees C, pH 8.0, or 37 degrees C, pH 5.5, in early stationary phase. At 37 degrees C, pH 8.0, inv is weakly expressed. To identify which gene(s) are required for inv regulation, we screened for transposon insertions that decreased expression of an inv-'phoA chromosomal reporter at 26 degrees C. Of 30 000 mutants screened, two were identified that had negligible inv expression in all conditions tested. Both of these independent mutants had an insertion into the same gene, designated rovA (regulator of virulence). RovA has 77% amino acid identity to the Salmonella typhimurium transcriptional regulator SlyA. Complementation with the wild-type rovA allele restores wild-type inv expression as monitored by Western blot analysis, tissue culture invasion assay and alkaline phosphatase assay. There is also a significant decrease in invasin levels in bacteria recovered from mice infected with the rovA mutant; therefore, RovA regulates inv expression in vivo as well as in vitro. In the mouse infection model, an inv mutant has a wild-type LD50, even though the kinetics of infection is changed. In contrast, the rovA mutant has altered kinetics, as well as a 70-fold increase in the LD50 compared with wild type. Furthermore, because the rovA mutant is attenuated in the mouse model, this suggests that RovA regulates other virulence factors in addition to inv. Analysis of other proposed virulence factors such as Ail, YadA and the Yop proteins shows no regulatory role for RovA. The more severe animal phenotype combined with the lack of impact on known virulence genes aside from inv suggests RovA regulates potentially novel virulence genes of Y. enterocolitica during infection. PMID- 10672190 TI - A NapC/NirT-type cytochrome c (NrfH) is the mediator between the quinone pool and the cytochrome c nitrite reductase of Wolinella succinogenes. AB - Wolinella succinogenes can grow by anaerobic respiration with nitrate or nitrite using formate as electron donor. Two forms of nitrite reductase were isolated from the membrane fraction of W. succinogenes. One form consisted of a 58 kDa polypeptide (NrfA) that was identical to the periplasmic nitrite reductase. The other form consisted of NrfA and a 22 kDa polypeptide (NrfH). Both forms catalysed nitrite reduction by reduced benzyl viologen, but only the dimeric form catalysed nitrite reduction by dimethylnaphthoquinol. Liposomes containing heterodimeric nitrite reductase, formate dehydrogenase and menaquinone catalysed the electron transport from formate to nitrite; this was coupled to the generation of an electrochemical proton potential (positive outside) across the liposomal membrane. It is concluded that the electron transfer from menaquinol to the catalytic subunit (NrfA) of W. succinogenes nitrite reductase is mediated by NrfH. The structural genes nrfA and nrfH were identified in an apparent operon (nrfHAIJ) with two additional genes. The gene nrfA encodes the precursor of NrfA carrying an N-terminal signal peptide (22 residues). NrfA (485 residues) is predicted to be a hydrophilic protein that is similar to the NrfA proteins of Sulfurospirillum deleyianum and of Escherichia coli. NrfH (177 residues) is predicted to be a membrane-bound tetrahaem cytochrome c belonging to the NapC/NirT family. The products of nrfI and nrfJ resemble proteins involved in cytochrome c biogenesis. The C-terminal third of NrfI (902 amino acid residues) is similar to CcsA proteins from Gram-positive bacteria, cyanobacteria and chloroplasts. The residual N-terminal part of NrfI resembles Ccs1 proteins. The deduced NrfJ protein resembles the thioredoxin-like proteins (ResA) of Helicobacter pylori and of Bacillus subtilis, but lacks the common motif CxxC of ResA. The properties of three deletion mutants of W. succinogenes (DeltanrfJ, DeltanrfIJ and DeltanrfAIJ) were studied. Mutants DeltanrfAIJ and DeltanrfIJ did not grow with nitrite as terminal electron acceptor or with nitrate in the absence of NH4+ and lacked nitrite reductase activity, whereas mutant DeltanrfJ showed wild-type properties. The NrfA protein formed by mutant DeltanrfIJ seemed to lack part of the haem C, suggesting that NrfI is involved in NrfA maturation. PMID- 10672191 TI - Ascaris lumbricoides infection is associated with protection from cerebral malaria. AB - Following reports of increased IgE in severe malaria and hypothesizing that helminth coinfections could modify its outcome, we conducted a retrospective case control study to establish whether helminths affect the evolution of Plasmodium falciparum malaria. Some 182 severe cases, 315 mild controls and 40 controls with circulating schizonts were examined for intestinal helminths. Comparing cerebral malaria with mild controls, Ascaris lumbricoides was associated with a protective adjusted odds ratio (OR) of 0.58 (0. 32-1.03) P = 0.06, for coinfection with Ascaris and Necator americanus, OR = 0.39 (0.17-0.88) P = 0.02. Protection followed a dose-effect trend (P = 0.008). When comparing cerebral malaria cases and controls with circulating schizonts the OR was 0.25 (0.009-0.67) P = 0.006. We hypothesized that Ascaris infected patients may have had decreased cyto adherence, possibly through endothelial cell receptor downregulation and/or decreased splenic clearance leading to the absence of selection of virulent P. falciparum strains. IgE-anti-IgE immune complexes resulting from helminth preinfection may have an important role in influencing clinical presentation of severe malaria, and in establishing malaria tolerance, through the CD23/NO pathway. PMID- 10672192 TI - Secretion of IL-12 by murine macrophages activated by immunoglobulin receptor mediated internalization of the surface coat of Trichinella spiralis larvae. AB - Trichinella spiralis larvae incubated with a rabbit antiserum raised against the larval surface coat bound murine macrophages to the parasite surface. Cell binding was not observed without the antisurface coat serum, or with incubation of larvae in normal rabbit serum, or with antibodies to keyhole limpet haemocyanin which identify a cryptic T. spiralis larval antigen. Cell adherence to the larval surface was lost by treatment of the cells with the lysosomotropic drug primaquine, implicating a receptor-mediated mechanism. Cells adhering to the parasite surface internalized parasite surface coat material, which was subsequently concentrated into endosomes. Culture supernatants from these cells contained enhanced levels of IL-12. Thus, the initial Th1 response to T. spiralis infection may be explained by these data. PMID- 10672193 TI - Costimulatory signal through CD86 is important in Th2 response in Trichinella spiralis infected mice. AB - In order to study the role of the costimulatory signals in Th2 cytokine production, monoclonal antibodies (mAbs) against cell adhesion molecules (CAMs) were added to cultured cells obtained from the mesenteric lymph nodes (MLNs) of mice infected with Trichinella spiralis, followed by a determination of interleukin (IL)-5 and IL-4 in the culture supernatant. IL-5 production by MLN cells stimulated with somatic antigen was significantly reduced by addition of anti-CD86 but not by anti-CD80 mAb. Combination of anti-CD80 and anti-CD86 mAbs reduced IL-5 production most effectively. IL-4 production induced by anti-CD3 mAb was suppressed only by the addition of anti-CD86 mAb. Blockade of the ICAM-1/LFA 1 and VCAM-1/VLA-4 interactions was less effective on the production of IL-5 and IL-4 than the addition of anti-CD86 mAb alone. In contrast to the in vitro cytokine production, intraperitoneal injection of anti-CD80, anti-CD86 mAb, or both, similarly suppressed the peak of the eosinophilia on day 21. Elevation of somatic antigen-specific IgE and IgG1 levels as well as total IgE was not inhibited by the administration of anti-CD80, anti-CD86 mAb or both. In-vitro and in-vivo effects of CTLA-4 immunoglobulin were similar to those of combined treatment with anti-CD80 and anti-CD86 mAbs. These results suggest that the interaction between antigen-presenting cells and CD4 T cells through CD86 are most important in Th2 response during T. spiralis infection. PMID- 10672195 TI - Relationship between parasite-specific antibody responses and intensity of Opisthorchis viverrini infection in hamsters. AB - The kinetics of parasite-specific antibody responses in relation to worm burden and egg output were investigated in hamsters infected with 25, 50 and 100 Opisthorchis viverrini metacercariae (MC). Levels of antibody to egg, excretory secretory (ES) and somatic antigens were examined by ELISA on days 1, 3, 7, 14 and month 1 postinfection (p.i.), and repeated monthly up to 6 months. The antibody responses were first detected as early as 14 days after infection. Hamsters that were infected with 100 MC and 50 MC showed higher antibody levels than those of 25 MC, during early infection until 1 month p.i. Then, the antibody levels were increased rapidly to a plateau at approximately month 2 p.i. and, subsequently, were relatively stable in all groups. The average antibody levels to egg and somatic, but not to ES antigens, were significantly higher in hamsters infected with 25 MC than those of 50 MC and 100 MC. These antibody responses, particularly to egg and ES antigens, were not correlated with worm burden or egg output. Overall, higher antibody responses were found in the order: ES, somatic and egg antigens. The significant lower antibody responses in chronic and heavy infections than those with mild infection may a result of immunosuppression. PMID- 10672194 TI - Vaccination of Manchego lambs against Haemonchus contortus with a somatic fraction (p26/23) of adult parasites. AB - A low molecular weight fraction from adult Haemonchus contortus containing two peptides (p26/23) was used to vaccinate Manchego female lambs between 3.5 and 5 months of age. Immunizing injections were given three times on days 0, 14 and 28 of the experiment. On day 43, lambs were challenged with 400 third stage larvae/kg live weight. Vaccination induced a lengthening of prepatent periods, significant reduction (> 60%) in mean faecal egg counts and smaller variations in packed cell volume values. At necropsy, average worm burden in the vaccinated lambs was significantly lower (61.6%) than that found in unvaccinated challenged animals. A clear correlation was found between protection and serum antibody response in immunized lambs. PMID- 10672196 TI - Fasciola hepatica infection downregulates Th1 responses in mice. AB - Immune responses induced with helminth parasites have been extensively studied, but there is limited information on those to Fasciola hepatica, especially on the subtype of T cell induced with this parasite. We investigated the local and systemic T cell responses of different strains of mice following oral infection with doses of metacercariae from F. hepatica. Spleen cells from BALB/c and 129Sv/Ev mice given a low-dose (5 metacercariae) infection exhibited a Th2 response, producing high levels of the cytokines IL-4 and IL-5, and low levels of IFN-gamma and IL-2. In contrast, C57BL/6 mice showed a mixed Th1/Th2 response. A more marked polarization to a Th2 response was observed in BALB/c, 129Sv/Ev exposed to a high-dose (15 metacercariae) infection and the C57BL/6 mice also exhibited a clear Th2 response. IL-4 defective (IL-4-/-) C57BL/6 mice infected with 5 metacercariae produced less IFN-gamma and more IL-5 compared to their wild type C57BL/6 counterparts, suggesting that IL-4 is important in establishing the Th2 type response in murine fasciolosis. However, the secretion of IFN-gamma and IL-2 was completely suppressed in the high-dose infection and this was also observed in IL-4-/- mice. Thus, liver flukes may secrete molecules that downregulate Th1 responses. T cell responses in the mesenteric (MLN) and hepatic lymph nodes (HLN) were also examined since newly excysted juveniles infect through the intestinal wall of their host before migrating to the hepatic tissue. Cells from both MLN and HLN secreted higher levels of IL-4 and IL-5 compared to spleen cells. We also observed a difference in cytokine profiles secreted by the MLN and HLN, which may reflect responses to antigens liberated by newly excysted juveniles and hepatic stage parasites, respectively. PMID- 10672197 TI - Interferon-gamma-independent CD8+ T cell-mediated protective anti-malaria immunity elicited by recombinant adenovirus. AB - Recombinant adenovirus, expressing the CS protein of Plasmodium yoelii, AdPyCS, was shown to induce a comparable degree of T cell-mediated protection against malaria as a single dose of irradiated P. yoelii sporozoites, causing inhibition of liver stage development. We now report that differently from sporozoite induced immunity, interferon (IFN)-gamma does not mediate the protective immunity induced by AdPyCS, since a similar degree of protection was observed in AdPyCS immunized mice lacking IFN-gamma-/- and the IFN-gamma receptor (IFN-gammaR-/-) compared to that in wild-type mice. Depletion of CD8+ T cells from these immunized mice almost completely abolished the AdPyCS-induced immunity, indicating that the immunization with AdPyCS induces CD8+ T cell-mediated protective anti-malaria immunity, which is independent of IFN-gamma. PMID- 10672198 TI - Do we negotiate human health? PMID- 10672199 TI - Reliability of malaria microscopy in epidemiological studies: results of quality control. AB - To assess the interrater reproducibility of malaria microscopy in epidemiological studies, 711 thick blood films from population-based surveys were randomly selected and reread by 4 experienced microscopists. Sample estimates of the prevalence of P. falciparum infection, geometric mean parasite density and the proportion of samples above various parasite density cut-off levels were almost identical in the routine and quality control readings. Differences were, however, encountered in the sample estimates for gametocyte ratio, proportion of mixed infection and average density index. In all three cases the quality control result was significantly higher than the routine evaluation. On the level of the individual slide there was good interrater agreement for the presence of P. falciparum infections (Kappa index kappa = 0.79) which was even better when parasite densities between 4 and 100/microl were excluded (kappa = 0.94). With respect to the assessment of parasite density, a high level of disagreement was found. While the mean difference between the two readings was not different from 0, the second reading was between 0.12 and 10 times that of the first. However, the level of disagreement significantly fell with increasing parasite densities. Thus malaria microscopy is very reliable for the estimation of parasite ratios and geometric mean parasite densities within and between studies as long as the same methodology is used, but tends to underestimate the gametocyte ratio and proportion of mixed infections. Care must be taken, however, when individual parasite density is related to other explanatory variables, due to the high degree of variability in the parasite enumeration. PMID- 10672200 TI - Severe anaemia in Zambian children with Plasmodium falciparum malaria. AB - BACKGROUND: Severe anaemia and cerebral malaria are highly prevalent complications of Plasmodium falciparum malaria among African children. The mechanisms of severe malarial anaemia, and the relative importance of this condition in comparison to cerebral malaria, are not known for many regions of Africa. METHODS We reviewed the records of 6200 children up to 6 years of age admitted to one rural Zambian hospital between 1994 and 1996. Severe malarial anaemia was defined as an haemoglobin concentration < 5.0 g/dl in a patient with asexual forms of P. falciparum in the peripheral blood. Cerebral malaria was defined as impaired consciousness (Blantyre coma score < 5) not attributable to any other cause in a patient with a positive malaria smear. RESULTS Severe malarial anaemia was found in 590 children (9.5% of paediatric admissions) and strictly defined cerebral malaria occurred in 286 children (4.6% of paediatric admissions); 98 of these patients had the combination of both complications. Severe malarial anaemia correlated strongly with the degree of parasitaemia, with malnutrition as indicated by low weight for age, with absence of fever and with presentation late in the malaria season. In comparison, patients with cerebral malaria were more often febrile and presented earlier in the malaria season. The case fatality rate of severe malarial anaemia (0.088) was about half that of cerebral malaria (0.189), but because severe malarial anaemia was more common, these two forms of complicated malaria were implicated in similar numbers of in hospital paediatric deaths. CONCLUSION Severe anaemia is a more common complication of P. falciparum malaria in hospitalized Zambian children than cerebral malaria and is associated with a similar number of deaths. Malnutrition and changes in immune response patterns due to prolonged exposure to P. falciparum may contribute to the development of this complication. PMID- 10672201 TI - Malaria prevalence and a brief entomological survey in a village surrounded by rice fields in Khammouan province, Lao PDR. AB - We surveyed Nongceng, a village in a south-eastern province of Lao PDR, for malaria and its vectors. Nongceng is situated in a basin and surrounded by rice fields. In February 1998 (dry season), 28.6% of 126 villagers were infected with malaria, and in September 1998 (rainy season), 16.3% of 147 villagers. The prevalence of malaria infection was consistently high in children under 10, and the predominant malaria species was Plasmodium falciparum. In brief surveys of the mosquitoes performed on the same day as the malaria surveys, 2007 Anopheles females from 12 species were collected by means of human bait, animal bait and resting collections. Of the vector species known to be important in transmitting malaria in neighbouring Thailand - An. minimus, An. dirus, and An. maculatus groups - only An. minimus was found. Its density was, however, very low in both seasons and it was therefore unlikely to be the vector. In fact, An. nivipes accounted for more than 65% of all mosquitoes collected and was the most common species collected from human baits. The results of this study show that endemic areas of malaria in Lao PDR are not necessarily related to forest. Rather, An. nivipes is suspected to be the most important vector. PMID- 10672202 TI - Domestic hygiene and diarrhoea - pinpointing the problem. AB - Improving domestic hygiene practices is potentially one of the most effective means of reducing the global burden of diarrhoeal diseases in children. However, encouraging behaviour change is a complex and uncertain business. If hygiene promotion is to succeed, it needs to identify and target only those few hygiene practices which are the major source of risk in any setting. Using biological reasoning, we hypothesize that any behaviours which prevent stools from getting into the domestic arena, the child's main habitat, are likely to have a greater impact on health than those practices which prevent pathogens in the environment from being ingested. Hence safe stool disposal, a primary barrier to transmission, may be more important than hand-washing before eating, which constitutes a secondary barrier, for example. We review the epidemiological evidence for the effect of primary and secondary barrier behaviours and suggest that it supports this conclusion. In the absence of local evidence to the contrary, hygiene promotion programmes should give priority to the safe disposal of faecal material and the adequate washing of hands after contact with adult and child stools. PMID- 10672203 TI - Validity of data-derived algorithms for ascertaining causes of adult death in two African sites using verbal autopsy. AB - background Verbal autopsy (VA) is used to ascertain causes of death using information obtained from bereaved relatives. Causes of death can be ascertained from VA questionnaires by a panel of physicians or from predefined algorithms. In a previous study, we developed data-derived algorithms using VA data from 796 adult deaths in hospitals in Tanzania, Ethiopia, and Ghana (primary sites). These computerized algorithms accurately estimated the cause-specific mortality fractions (CSMFs) for deaths due to injuries, meningitis, TB/AIDS and diarrhoeal diseases in the primary sites. Since the same data were used to generate and to validate the algorithms, the accuracy of our algorithms may have been overestimated. We report here on the validity of the algorithms when they were applied to VA data from two secondary sites in Ghana and Tanzania. Here, 'validity' is taken to mean the degree to which the algorithms replicated the physician-generated CSMF for major causes of death, when applied to the same VA data. methods VA interviews were conducted in two secondary sites: in Navrongo, Ghana, on 406 adult deaths, where three local physicians independently reviewed the questionnaires and assigned a cause of death. In Morogoro, Tanzania, VA interviews were conducted on 209 adult deaths, and a panel of physicians independently reviewed the VA questionnaires together with the hospital death certificates or hospital records to determine the cause of death. The CSMF obtained using each algorithm was compared with the CSMF obtained using physician review. results For injuries and meningitis, the algorithms and physician review estimated a similar CSMF in the Morogoro and Navrongo data. For TB/AIDS, the algorithm estimated a similar CSMF as the physicians in Morogoro. The algorithm for diarrhoeal diseases did not agree closely with the physicians in Morogoro or Navrongo. conclusions In general, our data-derived algorithms for assigning causes of death due to injuries, meningitis, and TB/AIDS estimated a similar CSMF as the physicians in the secondary sites. Recommendations for further validation and refinement are discussed. Computerized algorithms offer a potentially quick, affordable, and feasible method for assigning causes of death in mortality surveillance or studies using VA. PMID- 10672204 TI - HIV heterogeneity and proximity of homestead to roads in rural South Africa: an exploration using a geographical information system. AB - objective To describe heterogeneity of HIV prevalence among pregnant women in Hlabisa health district, South Africa and to correlate this with proximity of homestead to roads. methods HIV prevalence measured through anonymous surveillance among pregnant women and stratified by local village clinic. Polygons were created around each clinic, assuming women attend the clinic nearest their home. A geographical information system (GIS) calculated the mean distance from homesteads in each clinic catchment to nearest primary (1 degrees ) and to nearest primary or secondary (2 degrees ) road. results We found marked HIV heterogeneity by clinic catchment (range 19-31% (P < 0.001). A polygon plot demonstrated lower HIV prevalence in catchments remote from 1 degrees roads. Mean distance from homesteads to nearest 1 degrees or 2 degrees road varied by clinic catchment from 1623 to 7569 m. The mean distance from homesteads to a 1 degrees or 2 degrees road for each clinic catchment was strongly correlated with HIV prevalence (r = 0.66; P = 0.002). conclusions The substantial HIV heterogeneity in this district is closely correlated with proximity to a 1 degrees or 2 degrees road. GIS is a powerful tool to demonstrate and to start to analyse this observation. Further research is needed to better understand this relationship both at ecological and individual levels, and to develop interventions to reduce the spread of HIV infection. PMID- 10672205 TI - Acceptance and use of communal filtration units in guinea worm eradication. AB - The use of cloth to filter drinking water for guinea worm prevention is a long standing control strategy and part of a mixed approach that includes the provision of wells, chemical treatment of ponds and protection of water supplies. As the goal of eradication nears, filters are a useful component of the quick response needed to implement case containment at village level. Various designs of filters have been used. Individual hand-sewn filters (HSFs) using monofilament nylon cloth have played a central role in village-based control to date. Problems such as the need to continually reinforce correct habitual filtering behaviour have led to the design and testing of communal filtration units (CFUs) made from metal oil drums with filter cloth inserted in the top and spigots at the bottom. Approximately one year after the introduction of CFUs in the South-western Zone of Nigeria, village surveys were conducted to determine opinions about the two types of filters and reported use. Percentage use was calculated by dividing the number of times water was filtered in the week preceding the survey by the number of times water was collected in that week. Those respondents with access to CFUs filtered an average of 91.9% of the time compared to 75.7% of those with HSFs. Using the village as level of analysis since it was the main level of intervention, the average percent of times villagers in CFU villages filtered was 91.1% compared to 77.8% in HSF villages. Although CFUs were more expensive in the short run, their greater acceptance by villagers is a factor to recommend their wider implementation to speed up elimination of guinea worm from Nigeria. PMID- 10672206 TI - Control of urinary schistosomiasis: an investigation into the effective use of questionnaires to identify high-risk communities and individuals in Niger State, Nigeria. AB - Schistosomiasis is a public health problem in Nigeria. Although there is a national programme for its control, there is the need for reliable and simple means of rapidly diagnosing communities to provide a detailed map on the distribution of the disease in the country, in order to prioritize control activities, as well as to monitor the effectiveness of control operations. A rapid assessment technique using school questionnaires was tested in Borgu Local Government Area (LGA), Niger State, north-western Nigeria. Following a series of focus group discussions, the questionnaires were adapted before they were administered through the school system to 60 primary schools in Borgu LGA. Correctly completed questionnaires were returned from 58 schools (97%) within 4 weeks. Questionnaires were validated by reagent stick tests performed by trained teachers. Their results proved to be reliable compared to those obtained by our research team in 20 randomly selected schools. Overall prevalences of microhaematuria at 1+ and 2+ levels were 45.7% and 27. 1%, respectively. Highly significant correlations were obtained between school prevalence of microhaematuria and reported schistosomiasis, as well as reported blood in urine. The diagnostic performance of the questionnaires at the 2+ level of microhaematuria was very good. The design of our study also allowed data analysis on an individual level, and multivariate analysis revealed highly significant odds ratios for reported schistosomiasis and reported blood in urine to detect an individual with urinary schistosomiasis. Our results are in good agreement with reports from other African countries, and questionnaires can be recommended for rapid identification of communities at highest risk of urinary schistosomiasis in Nigeria, so that scarce resources of the national control programme can be used most effectively. PMID- 10672207 TI - Development of rapid assessment procedures for the delimitation of lymphatic filariasis-endemic areas. AB - Lymphatic filariasis caused by Wuchereria bancrofti is a major public health problem in 73 tropical and subtropical countries including India. Delimitation of endemic areas is essential to plan control operations. The current method of night blood survey (NBS) for delimitation is cumbersome, time-consuming and expensive. Therefore, there is a need to develop assessment procedures which can rapidly delimit endemic areas. For this purpose we evaluated three procedures: direct interviewing of key informants using structured questionnaires, an indirect method of a self-administered questionnaires to key informants and physical examination by health workers for the presence of chronic filarial disease. Thirty rural communities in a filariasis-endemic region in Cuddalore district in Tamil Nadu State in southern India constituted the study population. The determination of filariasis endemicity in the village communities assessed by the above procedures was compared in terms of rapidity, specificity, sensitivity and cost with the microfilaria rate and disease rate obtained by night blood sample survey and clinical examination by physicians. Prevalence score, control preference score and weighted mean number of cases with filarial disease per village were calculated using the key informant questionnaire techniques. While the prevalence and control preference score showed low sensitivity and moderate specificity, weighted mean number of cases showed high sensitivity and moderate specificity in identifying endemic villages. The prevalence of disease as determined by the physical examination of a sample population by health workers was highly sensitive in identifying communities endemic for filariasis. The degree of association between the disease rates estimated by physician and trained health workers was significant (r = 0.56; P < 0.05). These observations suggest that the weighted mean number of cases per village obtained through key informant techniques may be considered at a primary level to crudely identify endemic areas, followed by physical examination by health workers for filariasis, since it is relatively cheap and rapid. PMID- 10672208 TI - Detection of praziquantel resistance in schistosomes. PMID- 10672209 TI - [Diversity of feeding behavior of Glossina palpalis palpalis in the forest belt of the Ivory Coast: relation to the prevalence of human African trypanosomiasis]. AB - The feeding habits of Glossina palpalis palpalis, the main vector of human African trypanosomiasis (HAT) were retrospectively analysed using data collected between 1984 and 1994 in five areas in the forest belt in the mid-west of Cote d'Ivoire. The authors compare the feeding habits of the vector in these different foci. This analysis is aimed at determining if there is any relationship between the feeding pattern of tsetse-flies and the prevalence rates of HAT. The feeding pattern was measured using two indices: the conventional index of Shannon and Weaver (Ish) and a new one, the zoophily/anthropophily index (Za). The latter is an estimate of the ratio of the percentage of animal blood meals divided by the percentage of human blood meals. There was no correlation between apparent density and prevalence rate. A high Ish and a high Za were observed in the foci of Vavoua, Zoukougbeu and Sinfra where prevalence rates of HAT were high. Conversely, a low Ish and a low Za were observed in the hypoendemic areas of Daniafla and Gagnoa. Both indices are highly but not significantly correlated with prevalence rates. The Za index seemed to be more strongly correlated to the disease rate as compared to the Ish index. The epidemiological significance of these observations is discussed. PMID- 10672210 TI - Vertical dimension is a compounding problem. PMID- 10672211 TI - Eruption Guidance Appliance effects in the treatment of Class II, Division 1 malocclusions. AB - The objective of this research was to cephalometrically evaluate the possible effects of the Eruption Guidance Appliance on the craniofacial complex in a sample of 30 patients, over a treatment period of 26 months. The experimental sample consisted of 30 patients (13 females and 17 males), 27 of which presented with a Class II, Division 1 malocclusion and 3 with a Class I malocclusion. The mean initial chronologic age was 9 years; the treatment period lasted 26 months. A control group was used for comparison and consisted of 30 subjects (13 females and 17 males) of similar ages and spanned a similar observation period. Twenty six subjects of this control group had Class II, Division 1 malocclusions, and 4 had Class I malocclusions. Lateral cephalometric headplates were obtained for the experimental group initially and after 26 months of treatment. The subjects in the control group were randomly selected from a serial growth study sample from the Orthodontic Department at Bauru Dental School, University of Sao Paulo, for whom cephalometric headplates were obtained annually from 4 to 18 years of age. Comparative statistics were used to assess possible differences between the experimental and control groups during the 26-month period of observation. Results demonstrated statistically significant increases in mandibular growth, degree of mandibular protrusion, lower anterior and total anterior face height, mesial migration of the lower molars, and mandibular posterior dentoalveolar height. There was also lingual tipping and retrusion of the upper incisors, linear protrusion of the lower incisors, improvement in the maxillomandibular relationship and in molar relationship, as well as a significant decrease in the overjet and overbite and an inhibition of the vertical development of the upper incisors. The study demonstrated no significant changes in maxillary growth during the evaluation period. It was concluded from these results that the effects of the Eruption Guidance Appliance during this time period were mostly dentoalveolar, with a smaller, but significant, skeletal effect. PMID- 10672212 TI - Two-year follow-up of distraction osteogenesis: its effect on mandibular ramus height in hemifacial microsomia. AB - Distraction osteogenesis has been used to lengthen the mandible in patients with hemifacial microsomia. Questions regarding soft tissue and skeletal growth after distraction osteogenesis have not been clearly elucidated in the literature. In this case report, a 2-year follow-up of distraction osteogenesis in a 7 year old boy is documented with lateral and posterior/anterior cephalometric analysis. The analysis was performed preoperatively and at specific postsurgical periods to evaluate the facial soft tissue and skeletal growth patterns. Objective analysis of this growing patient after distraction osteogenesis clearly demonstrates that the anteroposterior elongation of the mandible is relatively stable, whereas the vertical lengthening and soft tissue effects are minimal. Critical evaluation of other patients who have undergone distraction osteogenesis is needed to determine if this was an isolated incident or the expected result in similar patients. PMID- 10672213 TI - An inference modeling of human visual judgment of sagittal jaw-base relationships based on cephalometry: part I. AB - When there appears to be a contradiction between a cephalometric tracing and what is seen, the clinician tends to place more reliance on the appearance of the face in profile than on a formal cephalometric evaluation of the sagittal jaw relationship. With this in mind, we derived a multiple regression model with cephalometric variables to explain the visual influences that affect the subjective classification of sagittal jaw relationships made by experienced orthodontists. The regression model that showed the highest coefficient of determination, 0.90, was constructed from a linear combination of the angle ANB, the anteroposterior position of gnathion, the SN length, and the mandibular body length. The performance reliability of the inference system has been tested for 175 female adult cases and will be reported in Part II of this article. PMID- 10672214 TI - Cleft lip and palate management with maxillary expansion and space opening for a single tooth implant. AB - An adult Class I malocclusion with a unilateral cleft lip and palate is presented. The maxillary transverse deficiency was managed with orthopedic expansion and the missing lateral incisor with space opening, bone grafting, and single tooth implant. The mild maxillary retrognathia and deficient lip support was managed with dental compensation. PMID- 10672215 TI - Reconstruction of an alveolar cleft for orthodontic tooth movement. AB - Bone grafting to repair an alveolar cleft has long been an integral part of the treatment of persons with unilateral and bilateral clefts of the lip and alveolus. The presence of the cleft places a limitation on the orthodontist who would like to move teeth in the area of the cleft. Various grafting materials have been placed in alveolar clefts in an attempt to solve this problem. The case to be presented is a patient with a Class II, Division 2, malocclusion with a left unilateral alveolar cleft and a repaired cleft lip. Ten months after initiating orthodontic treatment, a free gingival graft procedure was performed because of insufficient vestibular depth and the narrow width of the keratinized attached gingiva at the left maxillary lateral and central incisor region. Two months after periodontal surgery, a mix of decalcified freeze-dried bone allograft and a granular bioactive glass graft material (1:1) were applied subperiostally on the buccal aspect of the edentulous cleft region. Six months later, the teeth adjacent to the grafted alveolar cleft were orthodontically moved into the edentulous area. The treatment results indicated that orthodontic, periodontal, and surgical interventions resulted in a successful closure of the alveolar cleft as well as improved periodontal conditions of the teeth adjacent to the cleft area. From the orthodontic point of view, tooth movement can be achieved successfully into a bone graft made of freeze-dried bone and bioactive glass. PMID- 10672216 TI - Effect of light-cure time on the initial shear bond strength of a glass-ionomer adhesive. AB - With the introduction of photosensitive (light-cured) restorative materials in dentistry, various methods were suggested to enhance the polymerization of these materials including layering and the use of more powerful light-curing devices. The purpose of this study was to determine the effects of increasing the light cure time on the initial shear bond strength (in the first half hour) of a resin modified glass-ionomer adhesive. Eighty-six teeth were divided into 4 groups according to either; (1) the adhesive system used, namely resin, reinforced glass ionomer, or composite, and (2) the light-cure time for the glass ionomer adhesive, namely 40, 45, and 50 seconds. The bonding approach followed the manufacturer's instructions unless otherwise specified. The results of the analysis of variance comparing the 4 experimental groups (F = 19.4) indicated the presence of significant differences between the groups (P =. 0001). In general, the shear bond strength was greater for the composite adhesive system (?x(-) = 5.2 +/- 2.9 MPa), followed by the 2 groups bonded with the resin-reinforced glass ionomer adhesive and light cured for 50 seconds (?x(-) = 3.8 +/- 1.1 MPa) and 45 seconds (?x(-) = 3.4 +/- 2.7 MPa). On the other hand, the shear bond strength was significantly lower for the group bonded with the glass ionomer adhesive and light cured for 40 seconds only (?x(-) = 0.4 +/- 1.0 MPa). The present findings indicated the following: (1) the resin-reinforced glass-ionomer adhesive has a significantly lower shear bond strength in the first half hour after bonding when compared to a composite resin adhesive; (2) the initial bond strength of the glass-ionomer adhesive was significantly increased by increasing the light-cure time for an additional 5 to 10 seconds; (3) the mean increase in the shear bond strength between 5 and 10 seconds of additional light curing was not significant but the variability was less with the longer cure time. PMID- 10672217 TI - Interarch tooth size relationships of 3 populations: "does Bolton's analysis apply?". AB - This study evaluates whether Bolton's interarch ratios extend across populations and genders. The data were derived from systematically collected preorthodontic casts of 180 patients, including 30 males and 30 females from each of 3 populations (black, Hispanic, and white). Forty-eight mesiodistal contact points were digitized on each model, and the lengths of the anterior, posterior, and overall arch segments were calculated. The results showed significant (P <.05) ethnic group differences in all 6 arch segment lengths and in all 3 interarch ratios. Whites displayed the lowest overall ratio (92.3%), followed by Hispanics (93.1%), and blacks (93.4%). The group differences were due primarily to the relationships between the posterior segments. The arch segments of males were significantly larger than females; the overall and posterior ratios were also significantly larger in males than in females. Multiple regression analyses showed that individual differences in the overall ratio were most closely associated with the size of the lower second premolar, followed by the upper lateral incisors, upper second premolars, and the lower central incisors. In combination, these 4 teeth explained approximately 50% of the variation in the overall ratio between subjects. We conclude that interarch tooth size relationships are population and gender specific. Bolton ratios apply to white females only; the ratios should not be indiscriminately applied to white males, blacks, or Hispanics. PMID- 10672218 TI - Human tooth movement in response to continuous stress of low magnitude. AB - Conventional orthodontic therapy often uses force magnitudes in excess of 100 g to retract canine teeth. Typically, this results in a lag phase of approximately 21 days before tooth movement occurs. The current project was undertaken to demonstrate that by using lower force magnitudes, tooth translation can start without a lag phase and can occur at velocities that are clinically significant. Seven subjects participated in the 84-day study. A continuous retraction force averaging 18 g was applied to 1 of the maxillary canines, whereas a continuous retraction force averaging 60 g was applied to the other. The magnitude was adjusted for each canine to produce equivalent compressive stresses between subjects. Estimated average compressive stress on the distal aspect of the canine teeth was 4 kPa or 13 kPa. The moment-to-force ratios were between 9 and 13 mm. Tooth movement in 3 linear and 3 rotational dimensions was measured with a 3-axis measuring microscope and a series of dental casts made at 1- to 14-day intervals. The results showed a statistical difference in the velocity of distal movement of the canines produced by the 2 stresses (P =.02). The lag phase was eliminated and average velocities were 0.87 and 1.27 mm/month for 18 and 60 g of average retraction force. Interindividual velocities varied as much as 3 to 1 for equivalent stress conditions. It was concluded that effective tooth movement can be produced with lower forces and that because loading conditions were controlled, cell biology must account for the variability in tooth velocities measured in these subjects. PMID- 10672220 TI - AAO foundation: A celebration of success: preliminary program of the 100th annual session, april 28 - may 3, 2000 PMID- 10672221 TI - George S. Uchiyama, DDS, MS, 1922 to 1999 PMID- 10672219 TI - The effect of tooth size discrepancy on occlusion: An experimental study. AB - The purposes of this experimental study are the following: (1) to compare the anterior and overall tooth size ratios reported by Bolton to values reported in epidemiologic studies, (2) to assess the accuracy of tooth size discrepancy measurements, (3) to investigate to what extent generalized tooth size discrepancy affects occlusion, (4) to investigate the effect of leveling the curve of Spee, and (5) to evaluate the effect of extraction therapy of 4 premolars on occlusion. For the first part of the study, Bolton's mean anterior and overall tooth size ratio (as well as the extraction values) were compared with calculations derived from 4 publications reporting mean mesiodistal tooth width by using the t test (P Leb was shifted to the left, indicating a shortened life span on the CD1 background. Furthermore, the CD1 < Sod2-/- > Leb mice develop metabolic acidosis at an early stage as was observed with CD1 < Sod2-/- > Cje. When Sod2tmlCje was placed on C57BL/6J (B6) background, the -/- mice were found to die either during midgestation or within the first 4 days after birth. However, when the B6 < Sod2 -/+ > Cje were crossed with DBA/2J (D2) for the generation of B6D2F2 < Sod2-/- > Cje mice, an entirely different phenotype, similar to that described by Lebovitz et al., was observed. The F2 Sod -/- mice were able to survive up to 18 days, and the animals that lived for more than 15 days displayed neurological abnormalities including ataxia and seizures. Their hearts were not as severely affected as were those of the CD1 mice, and neurological degeneration rather than heart defect appears to be the cause of death. PMID- 10672233 TI - The blood-brain barrier and cerebrovascular pathology in Alzheimer's disease. AB - The pathology of Alzheimer's disease (AD) is not limited to amyloid plaques and neurofibrillary tangles. Recent evidence suggests that more than 30% of AD cases exhibit cerebrovascular pathology, which involves the cellular elements that represent the blood-brain barrier. Certain vascular lesions such as microvascular degeneration affecting the cerebral endothelium, cerebral amyloid angiopathy and periventricular white matter lesions are evident in virtually all cases of AD. Furthermore, clinical studies have demonstrated blood-brain barrier dysfunction in AD patients who exhibit peripheral vascular abnormalities such as hypertension, cardiovascular disease and diabetes. Whether these vascular lesions along with perivascular denervation are coincidental or causal in the pathogenetic processes of AD remains to be defined. In this chapter, I review biochemical and morphological evidence in context with the variable but distinct cerebrovascular pathology described in AD. I also consider genetic influences such as apolipoprotein E in relation to cerebrovascular lesions that may shed light on the pathophysiology of the cerebral vasculature. The compelling vascular pathology associated with AD suggests that transient and focal breach of the blood-brain barrier occurs in late onset AD and may involve an interaction of several factors, which include perivascular mediators as well as peripheral circulation derived factors that perturb the endothelium. These vascular abnormalities are likely to worsen cognitive disability in AD. PMID- 10672234 TI - Neurological changes induced by stress in streptozotocin diabetic rats. AB - Previous studies from our laboratory demonstrated that chronic stress produces molecular, morphological, and ultrastructural changes in the rat hippocampus that are accompanied by cognitive deficits. Glucocorticoid impairment of glucose utilization is proposed as a causative factor involved in stress-induced changes. Current studies have examined the neurological changes induced by stress in rats with a preexisting strain upon their homeostatic load--namely, in streptozotocin (stz)-diabetic rats. Administration of stz (70 mg/kg, i.v.) produced diabetic symptoms such as weight loss, polyuria, polydipsia, hyperglycemia, and neuroendocrine dysfunction. Morphological analysis of hippocampal neurons revealed that diabetes alone produced dendritic atrophy of CA3 pyramidal neurons, an effect potentiated by 7 days of restraint stress. Analysis of genes critical to neuronal homeostasis revealed that glucose transporter 3 (GLUT3) mRNA and protein levels were specifically increased in the hippocampus of diabetic rats, while stress had no effect upon GLUT3 expression. Insulin-like growth factor (IGF) receptor expression was also increased in the hippocampus of diabetic rats subjected to stress. In spite of the activation of these adaptive mechanisms, diabetic rats subjected to stress also had signs of neuronal damage and oxidative damage. Collectively, these results suggest that the hippocampus of diabetic rats is extremely susceptible to additional stressful events, which in turn can lead to irreversible hippocampal damage. PMID- 10672235 TI - Functional brain imaging in the resting state and during activation in Alzheimer's disease. Implications for disease mechanisms involving oxidative phosphorylation. AB - In vivo brain imaging of patients with Alzheimer's disease (AD) using positron emission tomography (PET) demonstrates progressive reductions in resting-state brain glucose metabolism and blood flow in relation to dementia severity, more so in association than primary cortical regions. During cognitive or psychophysical stimulation, blood flow and metabolism in the affected regions can increase to the same extent in mildly demented AD patients as in age-matched controls, suggesting that energy delivery is not rate limiting. Activation declines with dementia severity, and is markedly reduced in severely demented patients. These results suggest that there is an initial "normal" functionally-responsive stage in AD, followed by a late less responsive stage. Studies of biopsied and postmortem brain indicate that the initial stage is accompanied by selective and potentially reversible down-regulation of the brain enzymes, including cytochrome oxidase, which mediate mitochondrial oxidative-phosphorylation. PMID- 10672236 TI - Cellular and molecular mechanisms underlying perturbed energy metabolism and neuronal degeneration in Alzheimer's and Parkinson's diseases. AB - Synaptic degeneration and death of nerve cells are defining features of Alzheimer's disease (AD) and Parkinson's disease (PD), the two most prevalent age related neurodegenerative disorders. In AD, neurons in the hippocampus and basal forebrain (brain regions that subserve learning and memory functions) are selectively vulnerable. In PD dopamine-producing neurons in the substantia nigra striatum (brain regions that control body movements) selectively degenerate. Studies of postmortem brain tissue from AD and PD patients have provided evidence for increased levels of oxidative stress, mitochondrial dysfunction and impaired glucose uptake in vulnerable neuronal populations. Studies of animal and cell culture models of AD and PD suggest that increased levels of oxidative stress (membrane lipid peroxidation, in particular) may disrupt neuronal energy metabolism and ion homeostasis, by impairing the function of membrane ion-motive ATPases and glucose and glutamate transporters. Such oxidative and metabolic compromise may there-by render neurons vulnerable to excitotoxicity and apoptosis. Studies of the pathogenic mechanisms of AD-linked mutations in amyloid precursor protein (APP) and presenilins strongly support central roles for perturbed cellular calcium homeostasis and aberrant proteolytic processing of APP as pivotal events that lead to metabolic compromise in neurons. Specific molecular "players" in the neurodegenerative processes in AD and PD are being identified and include Par-4 and caspases (bad guys) and neurotrophic factors and stress proteins (good guys). Interestingly, while studies continue to elucidate cellular and molecular events occurring in the brain in AD and PD, recent data suggest that both AD and PD can manifest systemic alterations in energy metabolism (e.g., increased insulin resistance and dysregulation of glucose metabolism). Emerging evidence that dietary restriction can forestall the development of AD and PD is consistent with a major "metabolic" component to these disorders, and provides optimism that these devastating brain disorders of aging may be largely preventable. PMID- 10672237 TI - Use of cytoplasmic hybrid cell lines for elucidating the role of mitochondrial dysfunction in Alzheimer's disease and Parkinson's disease. AB - There is substantial evidence of mitochondrial defects in neurodegenerative disorders such as Alzheimer's and Parkinson's diseases (AD and PD). We have probed the molecular implications of mitochondrial dysfunction in these diseases by transferring mitochondria from platelets obtained from disease and control donors into mitochondrial DNA-depleted recipient neuron-based cells (rho 0 cells). This process creates cytoplasmic hybrid (cybrid) cells where the mitochondrial DNA (mtDNA) from the donor is expressed in the nuclear and cellular background of the host rho 0 cell. Differences in phenotype between disease and control groups can thus be attributed to the exogenous mitochondria and mtDNA. Key methodological issues relating to this approach were addressed by demonstrating that recipient rho 0 cells have < 1 mtDNA copy/cell, and that exclusive repopulation with donor mtDNA occurs in cybrid cells. Further, we describe that sampling of heterogeneous cell populations is a valid approach for cybrid analysis. Our studies show that the focal respiratory chain defects reported in platelets of AD and PD cybrids can be recapitulated in AD and PD cybrids. In addition, both AD and PD cybrids display increased oxidative stress and perturbations in calcium homeostasis. These data suggest that the transfer of a mtDNA defect from disease donor platelets is the likely cause of the cybrid biochemical phenotype, and highlight the potential value of these cell lines as cellular disease models. PMID- 10672238 TI - Polyglutamine domain proteins with expanded repeats bind neurofilament, altering the neurofilament network. AB - Proteins with expanded polyglutamine (polyQ) repeats cause eight inherited neurodegenerative diseases. Nuclear and cytoplasmic polyQ protein is a common feature of these diseases, but its role in cell death remains debatable. Since the neuronal intermediate filament network is composed of neurofilament (NF) and NF abnormalities occur in neurodegenerative diseases, we examined whether pathologic-length polyQ domain proteins interact with NF. We expressed polyQ green fluorescent fusion proteins (GFP) in a neuroblast cell line, TR1. Pathologic-length polyQ-GFP fusion proteins form large cytoplasmic aggregates surrounded by neurofilament. Immunoisolation of pathologic-length polyQ proteins co-isolated 68 kD NF protein demonstrating molecular interaction. These observations suggest that polyQ interaction with NF is important in the pathogenesis of the polyglutamine repeat diseases. PMID- 10672239 TI - Bioenergetics in Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant inherited neurodegenerative disorder with relentless course and prototypical clinical symptoms. In 1993 HD was associated with an expanded CAG triplet repeat stretch on chromosome 4 in the coding region of its target protein, huntington. The length of the resulting polyglutamine++ extensions correlates with lower age of onset and a higher density of ubiquitin-positive neuronal intranuclear inclusions. Recently it has been proposed that mutant huntington induces progressive neuronal cell death by an apoptotic mechanism. There is strong evidence that disturbances in cellular energy homeostasis and oxidative damage contribute to neurodegeneration. This review will summarize and discuss the current concepts that point towards an involvement of free radical-induced oxidative stress, glutamate excitotoxicity and mitochondrial respiratory chain defects in pathogenesis of HD. PMID- 10672240 TI - An integrated strategy for evaluation of metabolic and oxidative defects in neurodegenerative illness using magnetic resonance techniques. AB - The number of physiologic and metabolic phenomena amenable to analysis using magnetic resonance (MR) techniques is increasing every year. MR techniques can now evaluate tissue parameters relevant to TCA cyclemetabolism, anerobic glycolysis, ATP levels, blood-brain barrier permeability, macrophage infiltration, cytotoxic edema, spreading depression, cerebral blood flow and volume, and neurotransmitter function. The paramagnetic nature of certain oxidation states of iron leads to the ability to map out brain function using deoxyhemoglobin as an endogenous contrast agent, and also allows for mapping of local tissue iron concentrations. In addition to these metabolic parameters, the number of ways to generate anatomic contrast using MR is also expanding; and in addition to conventional anatomic scans, mapping of axonal fiber tracts can also be performed using the anisotropy of water diffusion. A strategy for integration of these multifarious parameters in a comprehensive neurofunctional exam in neurodegenerative illness is outlined in this paper. The goals of the integrated exam, as applied to a given neurodegenerative illness, can be subdivided into three categories: etiology, natural history, and therapeutic end points. The consequences of oxidative stress and/or mitochondrial dysfunction are explored in the context of the various parameters that can be measured using the integrated MR exam. PMID- 10672241 TI - Apoptosis and necrosis in cerebrovascular disease. AB - Neuronal death following ischemic insults has been thought to reflect necrosis. However, recent evidence from several labs suggests that programmed cell death, leading to apoptosis, might additionally contribute to this death. We have used both in vitro and in vivo models to study the role of apoptosis in ischemic cell death. Some features of apoptosis (TUNEL staining, internucleosomal DNA fragmentation, sensitivity to cycloheximide) were observed following transient focal ischemia in rats. Brief transient focal ischemia was followed by delayed infarction more than 3 days later; this delayed infarction was sensitive to cycloheximide. A cycloheximide-sensitive component of neuronal cell death was also observed in cultured murine neocortical neurons deprived of oxygen-glucose in the presence of glutamate receptor antagonists. This presumed ischemic apoptosis was attenuated by caspase inhibitors, or by homozygous deletion of the bax gene. Neurons may undergo both apoptosis and necrosis after ischemic insults, and thus it may be therapeutically desirable to block both processes. PMID- 10672242 TI - Effects of ebselen, a glutathione peroxidase mimic, in several models of mitochondrial dysfunction. PMID- 10672243 TI - EDTA-induced monovalent fluxes through the Ca2+ uniporter in brain mitochondria. PMID- 10672244 TI - Signaling events in NMDA receptor-induced apoptosis in cerebrocortical cultures. PMID- 10672245 TI - In vitro and in vivo protein oxidation induced by Alzheimer's disease amyloid beta-peptide (1-42). PMID- 10672246 TI - Depolarization of in situ mitochondria by hydrogen peroxide in nerve terminals. PMID- 10672247 TI - Monoamine oxidase inhibits mitochondrial respiration. PMID- 10672248 TI - Enhanced acetate and glucose utilization during graded photic stimulation. Neuronal-glial interactions in vivo. PMID- 10672249 TI - Calcium overload triggers rod photoreceptor apoptotic cell death in chemical induced and inherited retinal degenerations. PMID- 10672250 TI - Molecular mechanisms of free radical production and protective efficacies of antioxidants in in vitro ischemia-reperfusion. PMID- 10672251 TI - A disturbance in the neuronal insulin receptor signal transduction in sporadic Alzheimer's disease. AB - Disturbances of glucose and energy metabolism are hypothesized as pathogenetic factors in sporadic dementia of Alzheimer type (SDAT). Insulin and is receptors play an important role in the regulation of brain glucose metabolism and neuronal growth. In postmortem brain cortex in SDAT, the densities of brain insulin receptors were decreased compared to adult controls, but were increased in relation to aged controls. Tyrosine kinase activity, a signal transduction mechanism common to insulin and IGF-1 receptors, was reduced in SDAT in comparison to middle-aged and age-matched control groups. The data are consistent with a neurotrophic role of insulin in the human brain and an upregulation of insulin receptors is SDAT brain as a compensatory mechanism, possibly due to impaired signal transduction mechanism. PMID- 10672252 TI - Modulation of presenilin-1 processing by nitric oxide during apoptosis induced by serum withdrawal and glucose deprivation. PMID- 10672253 TI - Metabolic and glutamatergic disturbances in the Huntington's disease transgenic mouse. PMID- 10672254 TI - Inhibition of the neuronal insulin receptor causes Alzheimer-like disturbances in oxidative/energy brain metabolism and in behavior in adult rats. PMID- 10672255 TI - Temporary axonal conduction block and axonal loss in inflammatory neurological disease. A potential role for nitric oxide? PMID- 10672256 TI - Maturational changes in rabbit brain phosphocreatine and creatine kinase. PMID- 10672257 TI - Neurotoxicity and oxidative damage of beta amyloid 1-42 versus beta amyloid 1-40 in the mouse cerebral cortex. AB - Senile plaques (SP), a neuropathological hallmark of Alzheimer's disease (AD), are characterized by extracellular accumulations of beta amyloid (A beta). SP predominantly contain A beta 42 with a small amount of associated A beta 40. We determined the neurotoxic properties of A beta 42 as compared to A beta 40 by injections into the frontal cortex of three month old C57BL/6 mice. A beta 42 was associated with a significantly larger area of glial fibrillary acidic protein (GFAP) immunoreactivity and a greater density of reactive astrocytes than A beta 40. Immunohistochemical staining for markers of oxidative damage against 3 nitrotyrosine (3-NT) and 8-hydroxydeoxyguanosine (8-OHDG) were significantly more intense around the A beta 42 injection compared to the A beta 40 injection sites. These findings are consistent with previous in vitro studies and suggest that A beta 42 is more neurotoxic and may generate more free radical damage than A beta 40. PMID- 10672258 TI - Ganglioside GD3, the mitochondrial permeability transition, and apoptosis. PMID- 10672259 TI - Interactions between melatonin, reactive oxygen species, and nitric oxide. AB - Accumulation of reactive oxygen species is critical for the neuropathology of Alzheimer's disease. Melatonin hormone, an antioxidant, could play a key role in aging and senescence. Nitric oxide, a biologically active unstable radical, is synthesized by nitric oxide synthase when converting L-arginine to L-citrulline. We have investigated whether the treatment of cultured cells with melatonin could possibly reduce the release of free radicals and other ROS. We assayed NO indirectly by measuring the level of its stable end products, nitrite/nitrate (NOx), using the Griess reagent. When the neuroblastoma cells such as N1E-115 were treated with a NO donor such as sodium nitroprusside (SNP), a significant level of NOx was detected in a time- and dose-dependent manner in the conditioned medium compared to the untreated cells or SNP-containing media. In neuroblastoma cells, the release of NOx as mediated by SNP was significantly inhibited by treatment with (i) carboxy-PTIO, a NO scavenger; (ii) SOD-1, superoxide dismutase; and (iii) melatonin. In these cells SNP-mediated NOx release was mediated by superoxide ions and/or free radicals that can be inhibited by melatonin. The ROS-scavenging function of melatonin along with its neuroprotective and neurodifferentiating role can be utilized for the prevention of neurodegenerative disorders such as AD. PMID- 10672260 TI - Effect of oxidative stress on DNA damage and beta-amyloid precursor proteins in lymphoblastoid cell lines from a Nigerian population. AB - The epsilon 4 allele of apolipoprotein E (APOE) is strongly associated with late onset Alzheimer's disease (AD) in Caucasian populations, but our studies suggest that APOE epsilon 4 is not a risk factor for AD in Nigerian blacks and is a weak risk factor in African-Americans. The prevalence of AD is lower in Nigerians than in African-Americans. Increased oxidative damage to macromolecules in brain tissue by reactive oxygen species (ROS) has been reported in AD. Here we examined the effects of endogenous and induced oxidative stress on total (nuclear and mitochondrial) DNA damage in lymphoblastoid cell lines (5 probable AD and 3 controls) from Ibadan, Nigeria. Cells were exposed to 200 microM t-butyl peroxide (a generator of ROS) for 4 hours. Total DNA was isolated and digested with nuclease P1 and alkaline phosphatase. DNA fragments were separated by HPLC and the levels of 8-hydroxy-2'-deoxyguanosine (OH8dG, an indicator of DNA damage) and deoxyguanosine (dG) determined. We did not detect a significant difference in the OH8dG/dG ratio in untreated or treated cell lines in the two groups, and this was independent of APOE genotype. We also examined, by Western blotting, the level of beta-amyloid precursor protein (APP) which is involved in AD. The level of the heat shock protein (HSP-70) was examined as a control. There was a slight decrease in levels of APP and HSP-70 following treatment. Studies in cell lines from Caucasian subjects have shown an increase in mitochondrial DNA damage following oxidative challenge. Our preliminary results suggest that African populations are less vulnerable to chemical-induced oxidative DNA damage. PMID- 10672261 TI - Evidence for energy failure following irreversible traumatic brain injury. PMID- 10672262 TI - Regulation of mitochondrial gene expression in differentiated PC12 cells. PMID- 10672263 TI - Does dopamine contribute to striatal damage caused by impaired mitochondrial function? PMID- 10672264 TI - Discordance between traditional pathologic and energy metabolic changes in very early Alzheimer's disease. Pathophysiological implications. AB - These results suggest that neither the loss of entorhinal efferents nor cholinergic deficit explains all the metabolic features seen in very early AD. Given recent immunohistological evidence of massive glutamatergic synaptic alteration in early AD cortex and insights into neuronal and glial mechanisms of glucose metabolism, very early metabolic changes in AD probably reflect a significant impairment of glycolytic activities in the cortico-cortical glutamatergic systems in a preclinical stage of the disease. However, the exact mechanisms of such impairment in these neurons are yet to be determined. PMID- 10672265 TI - Low frequencies of mitochondrial DNA mutations cause cardiac disease in the mouse. PMID- 10672266 TI - Effects of the solvent 1,2,4-trimethylcyclohexane on respiratory burst in human neutrophil granulocytes. A chemiluminescence and electron paramagnetic resonance spectrometry study. PMID- 10672267 TI - Neuronal RNA oxidation in Alzheimer's disease and Down's syndrome. PMID- 10672268 TI - Ca(2+)-dependent permeability transition and complex I activity in lymphoblast mitochondria from normal individuals and patients with Huntington's or Alzheimer's disease. PMID- 10672269 TI - Mitochondrial porin, a novel target to prevent ischemia-induced neurodegeneration? PMID- 10672270 TI - Bcl-2 and p53: role in dopamine-induced apoptosis and differentiation. AB - The fate of a neuron in the developing brain to multiply, differentiate, or die in an apoptotic manner depends on the expression of genes that are involved in regulating the cell cycle. Recent studies determined the involvement of several genes, including cyclin A and B2, in dopamine-induced apoptosis in cultured chick sympathetic neurons. Another gene that plays a role in apoptosis and differentiation of neurons, oligodendrocytes and PC12 cells is p53. It is also known that DNA damage increases p53 levels, triggering repair or apoptosis in response to moderate or severe damage, respectively. NMB cells express active and inducible forms of p53, thus being particularly suitable to analyze the role of this gene in dopamine-induced apoptosis and differentiation. The main observation of this work is that low concentrations of dopamine induce differentiation while high concentrations induce apoptosis, and that concentrations of dopamine that induce apoptosis increased p53 levels. There peak increase in p53 was within 3-6 h, before cell death. Thus, treatment with a high dopamine concentration may result in oxidation products and/or free radicals that heavily damage DNA, thus increasing p53 levels and initiating a cascade of events leading to apoptosis. Lower concentrations of dopamine apparently have a milder damaging effect on the DNA and induce growth arrest and differentiation. In various systems Bcl-2 inhibits cell death, being apoptotic or necrotic. Bcl-2, and other members of the family, such as Bax, are located downstream to p53 in the apoptotic pathway, and they contain negative or positive p53 response elements. Bcl-2 also protects cells by acting as antioxidant. Neuronal differentiation may be accompanied with an increase in Bcl-2, though it was suggested that the role of Bcl-2 in differentiation is less critical than in apoptosis. Herein, Bcl-2 was found to inhibit dopamine neurotoxicity. Whether the expression of Bcl-2 is regulated by different dopamine concentrations, or by dibutyryl-cAMP and DMSO, remains to be determined. PMID- 10672271 TI - Mitochondria mediate nitric oxide-induced cell death. PMID- 10672273 TI - Measurement of oxidative DNA damage in the human p53 and PGK1 gene at nucleotide resolution. PMID- 10672272 TI - Induction of apoptosis and necrosis in human neuroblastoma cells by cholesterol oxides. PMID- 10672274 TI - The glucose paradox in cerebral ischemia. New insights. AB - The present in vivo findings that lactate, accumulated during an ischemic episode, is an essential aerobic energy substrate during the initial postischemic period are in full agreement with out in vitro findings. Moreover, the beneficial effects of hyperglycemia are also in agreement with our and others' in vitro results that have demonstrated a neuroprotective effect of glucose against hypoxic change. The aggravation of ischemic delayed neuronal damage by glucose loading 15 min prior to the ischemic insult is likely the result of glucose induction of a short-acting (30 to 60 min) systemic factor (hormonal?) that, when combined with an ischemic insult, potentiates the ischemic damage. PMID- 10672275 TI - Interactions of chloromethyltetramethylrosamine (Mitotracker Orange) with isolated mitochondria and intact cells. PMID- 10672276 TI - Attenuation of neuronal death by NMDA and oxygen-glucose deprivation in cortical neurons maintained in high glucose. PMID- 10672277 TI - Astrocyte nitric oxide causes neuronal mitochondrial damage, but antioxidant release limits neuronal cell death. PMID- 10672278 TI - Mechanisms of selective neuronal cell death due to thiamine deficiency. AB - Multiple mechanisms contribute to the selective brain lesions observed in WKS and experimental thiamine deficiency. Recent evidence of early microglial activation and increased free radical production suggest that oxidative stress processes play an important early role in the brain damage associated with thiamine deficiency. PMID- 10672279 TI - Inhibition of alpha-ketoglutarate dehydrogenase due to H2O2-induced oxidative stress in nerve terminals. PMID- 10672280 TI - ATPases of synaptic plasma membranes and vesicles from rat cerebral cortex during aging and hypoxia. PMID- 10672281 TI - Mitochondrial compartmentation at the cellular level: astrocytes and neurons. PMID- 10672282 TI - Detection of respiratory chain defects in cultivated skin fibroblasts and skeletal muscle of patients with Parkinson's disease. PMID- 10672283 TI - Prevention of neurodegeneration by a neuroprotective radical scavenger. PMID- 10672284 TI - Mitochondria, neurodegenerative diseases, and selective neuronal vulnerability. PMID- 10672285 TI - Regional differences in mortality: a comparison between Austria, Hungary and Switzerland. AB - BACKGROUND: Important differences in mortality rates exist even between neighbouring countries. This should facilitate the identification of the lifestyle parameters underlying these differences. The mortality rates obtained in Hungary, Austria and Switzerland were compared. METHODS: The mortality rates for all-cause, total cardiovascular, total cancer and stroke mortality were obtained from a special tape from WHO. Nutritional data were obtained from FAO food balance sheets and from dietary surveys. Gompertz and polynomial equations were calculated from the age-specific mortality rates. FINDINGS: Great differences in mortality exist between the three countries. In the period from 1950 until 1995 mortality decreased in Austria and Switzerland, but increased in Hungary. In men, total cancer and total cardiovascular mortality also increased markedly in Hungary during the last 20 years. Hungary has a lower dietary P/S ratio, a higher level of animal/vegetal fat and a lower consumption of fruit than Austria and Switzerland, combined with a high level of salt consumption. The level of cigarette smoking is similar in Hungary and Switzerland. The increase in mortality rate in Hungary is less pronounced in women than in men. INTERPRETATION: The major differences in lifestyle between the three countries concern socio-economic and nutritional patterns. Epidemiological evidence favours nutrition as the most important determinant of the differences in mortality rates between the three countries. PMID- 10672286 TI - The effects of halothane and sevoflurane on QT dispersion. AB - QT dispersion is defined as the difference between QT (max) and QT (min) in the 12-lead surface ECG. It has been shown to reflect regional variations in ventricular repolarization and is significantly greater in patients with arrhythmic events than in those without them. The aim of this study was to examine the effects of halothane and sevoflurane on QT and QTc dispersion during inhalational induction of anaesthesia. The effects on QT and QTc dispersion of halothane and sevoflurane have been investigated during induction of anaesthesia. Forty-six ASA (American Society of Anaesthesiologists) physical status I-II patients, aged 16-50 years, undergoing general anaesthesia were randomly allocated to receive either halothane or sevoflurane. The mean baseline values for QT and QTc dispersion were not significantly different between the two groups (P > 0.05). QT dispersion was increased with halothane compared with baseline values (50 +/- 16 ms vs. 29 +/- 9 ms, P < 0.01) and after sevoflurane compared with baseline (48 +/- 15 vs. 33 +/- 8 ms, P < 0.01). Also, QTc dispersion was increased with halothane compared with baseline values (48 +/- 13 ms vs. 31 +/- 9 ms, P < 0.001) and after sevoflurane compared with baseline (50 +/- 14 vs. 40 +/- 11 ms, P < 0.01). The QTc interval did not change by both sevoflurane (443 +/- 7 vs. 431 +/- 21 ms, P > 0.05) and halothane (419 +/- 33 vs. 431 +/- 19 ms, P > 0.05) compared with baseline. Both halothane and sevoflurane cause myocardial repolarisation abnormalities in man in terms of increased QTc dispersion. This may be relevant in the aetiology of arrhythmias in patients during anaesthesia with halothane or sevoflurane. PMID- 10672287 TI - Structural heart adaptations in triathletes. AB - OBJECTIVE: To perform a triathlon in aerobic conditions, a variety of cardiovascular, haemodynamic and metabolic adaptations are required. The heart is the central concern and also the most important limiting factor. In this study we investigate the structural and functional heart adaptations of a group of triathletes. METHODS AND RESULTS: A group of 52 male triathletes was divided into 4 subgroups in function of their athletic results and compared with a control group of 22 healthy, very active but no athletic men. The groups had comparable anthropometric and general physical characteristics. Very significant differences in cardiac structure and cardiac function were observed between the groups. In the triathletes, we registered distinct signs of significantly mixed eccentric and concentric hypertrophy. Unlike the findings in a pathological left ventricular hypertrophy, the diastolic left ventricular function in triathletes was completely normal and even better than in the control group. The late passive diastolic filling period of the triathlete, in particular, seemed to have specific characteristics. The comparison between the subgroups of triathletes shows us that genetic factors probably play an important role in the cardiac adaptations in triathletes. CONCLUSIONS: In our opinion the "athletic heart" in triathletes is not a specific "physiological entity" but is a transitional phase to a dilated hypertrophic cardiomyopathy. Our study yields some arguments for the following proposition: "People are born as elite athletes, with specific characteristics of the left ventricle and with a specifically supernormal diastolic left ventricular function." PMID- 10672288 TI - Secular trends in cardiovascular mortality in Iran, with special reference to Isfahan. AB - OBJECTIVE: There has been a general decline in mortality from cardiovascular diseases (CVDs) in most of the developed countries since the beginning of the 1970s. Still, in recent years developing countries have seen an increasing frequency in CVD mortality. However, mortality rate studies in these populations are scarce. Here we report all-cause and CVD mortality rates for men and women aged 25-74 years over a 16-year period in 24 cities in Iran with special reference to the city of Isfahan. METHODS AND RESULTS: The study was based on national death records using the ninth international classification of diseases and age standardization was performed using the total population of Iran in 1985 as a standard. Due to limitations in available data, mortality rates for the specific categories of CVD for the whole country could not be provided. The in hospital death rates following myocardial infarction in coronary care units (CCUs) and cardiology departments in Isfahan hospitals were also assessed. The completed medical records from hospitals or the relatives of decedents were reviewed by physicians certified in internal medicine, cardiology and neurology to assess the reliability of death certificate data regarding CVD by determining the sensitivity and specificity of the death certificates against the standard of the reviewers. The official circulatory diseases proportional mortality ratio continues to rise since 1981 with a steep increase since 1987, constituting 26.6% and 47.3% of all deaths in 1981 and 1995, respectively. Age-adjusted all-cause and CVD mortality data were decreasing since 1981 and increasing since 1990. During those years age-adjusted CVD, stroke and other CVD mortality rates were decreasing in Isfahan with a slight increase in ischaemic heart disease (IHD) death rates in both sexes. Mortality rates based on sex showed a 38% and 24.8% decline in all-cause and CVD mortality in men between 1981 to 1995, and a 35% and 34.9% decline for female mortality rates for the same period, respectively. The in-hospital death rate following myocardial infarction in Isfahan was increasing between 1993 and 1995 with a slight decrease thereafter. The results of death certification assessment showed a specificity of 0.89 and a sensitivity of 0.43 with the positive and negative predictive values of 0.82 and 0.57, respectively. CONCLUSION: These data indicate that circulatory diseases remain a serious public health threat in Iran. It suggests the ongoing need for more regular, systematic and innovative surveillance data to improve the capability of measuring, explaining and predicting the disease trend on which the national public health policy depends. PMID- 10672289 TI - The effect of aminophylline infusion on the exercise capacity in patients with syndrome X. AB - Aminophylline was known to decrease effort angina and ischaemia in patients with coronary artery disease and stable angina. This effect has been explained by the antagonization of the vasodilatation caused by adenosine and the prevention of the transmural myocardial maldistribution (steal phenomenon). In this study, treadmill exercise tests (with Bruce protocol) were performed in 14 patients (11 women, 3 men, mean age 51 +/- 9.9 years) diagnosed with syndrome X (typical anginal chest pain, abnormal stress test, normal coronary angiography) before (basal condition) and after the acute i.v. aminophylline infusion (total dosage 6 mg/kg within 15 minutes). It was shown that aminophylline lengthened the time before the occurrence of ischaemia in patients with syndrome X by increasing the ischaemia threshold in spite of the occurrence of tachycardia but it had no effect on the total exercise duration. Aminophylline infusion also exerted a beneficial effect on exercise-induced chest pain. It has been suggested that the role of the adenosine receptor could be important in the pathogenesis of syndrome X. PMID- 10672290 TI - Implantation of 500 consecutive cardiac pacemakers in the electrophysiology laboratory. AB - OBJECTIVE: The purpose of this study was to examine the practice of cardiac bradycardia pacing in an electrophysiology laboratory and to evaluate the safety and efficacy of pacemaker implantation by cardiologists in this setting. METHODS AND RESULTS: We evaluated a consecutive series of the first 500 permanent pacemakers (mean age of patients: 74 +/- 14 years) implanted entirely in the electrophysiology laboratory at our institution. All procedures were performed by two cardiologists. Three-hundred and four patients (60.8%) received a dual chamber device. After 3 months of follow-up, procedure-related complications occurred in 19 patients (3.8%). The most frequent complication was lead dislodgement (1.4%). There were 2 postoperative infections (0.4%). The mean time to hospital discharge after the implant was 2.9 +/- 1.6 days. CONCLUSIONS: These results show that permanent pacemaker implantation may be successfully carried out by cardiologists in the electrophysiology laboratory with a low complication rate. Atrial lead dislodgement seems to be the most frequent complication in centres with a high proportion of dual-chamber pacemaker implantations. PMID- 10672291 TI - Prediction of peak exercise oxygen uptake by cardiopulmonary measurements at rest in heart transplant candidates. AB - OBJECTIVE: Peak oxygen uptake (VO2) is a powerful prognostic index, but maximal exercise testing in heart transplant candidates has a number of disadvantages. It is unknown whether it is possible to predict peak VO2 from a comprehensive dataset with parameters of heart and lung function at rest. METHODS: One hundred adult patients in sinus rhythm and with either idiopathic or ischaemic heart failure performed a graded cycle ergometer test until volitional fatigue and underwent radionuclide ventriculography, heart catheterization, and lung function measurements at rest. RESULTS: Weight, height, age, gender and aetiology of heart failure explained 48% of the variance of peak VO2. On top of these anthropometric, demographic and clinical patient characteristics, 12% of the variance of peak VO2 was additionally explained by all resting measurements combined, i.e. radionuclide left ventricular ejection fraction, peak ejection rate, peak filling rate, cardiac frequency, mean right atrial pressure, pulmonary capillary wedge pressure, pulmonary artery pressures, cardiac output, forced vital capacity, forced expiratory volume in one second, and pulmonary diffusing capacity (cumulative R2 = 0.60); among these, pulmonary vascular resistance was the most important predictor (+6%; P < 0.001). Analyses in a subset of 43 male patients pointed out that systemic pressures and vascular resistance were not related to peak VO2. CONCLUSION: On the basis of resting left ventricular function, haemodynamics, and routine pulmonary measurements, it is unlikely to accurately predict exercise tolerance in the majority of heart transplant candidates, i.e. patients with either idiopathic or ischaemic heart failure and able to exercise until exhaustion. PMID- 10672292 TI - Coexistence of a left posteroseptal tract with persistent left superior vena cava. Ablation through an anomalous superior vena cava. AB - A case of a young male with WPW syndrome due to a left posteroseptal tract associated with a persistent left superior vena cava is described. After unsuccessful ablation attempts with a number of different approaches at conventional target sites, the accessory connection was successfully ablated within the coronary sinus. This was achieved only when the ablation catheter was introduced through the persistent left superior vena cava. PMID- 10672293 TI - Reversible hypertrophic cardiomyopathy in congenital hyperinsulinism. AB - A neonate with congenital focal hyperinsulinism developed hypertrophic cardiomyopathy that progressed until partial pancreatectomy was performed. Surgery was followed by complete resolution of the condition. Hyperinsulinism may be a cause of treatable cardiomyopathy. PMID- 10672294 TI - Transoesophageal echocardiographic diagnosis of a dissected aorticovenous anastomosis causing cardiac tamponade. PMID- 10672295 TI - Intra-operative premature rupture of the cerebral aneurysms. Analysis of the causes and management. AB - The causes and management of intra-operative premature rupture are analysed and discussed. During the past 6 years, the authors, performed 398 consecutive direct surgical interventions for ruptured cerebral aneurysms. Intra-operative premature rupture is defined as a rupture which occurs before the securing of the parent arteries or the neck of the aneurysm and is out of control, at least temporarily. The causes and management were retrospectively analyzed by reviewing video tape recordings. Intra-operative premature ruptures which met the above definition occurred in 24 cases (6.0%). The causes were as follows: 1.) dural opening and arachnoid opening (8.3%), 2.) haematoma removal (12.5%), 3.) brain retraction (16.7%), 4.) aneurysm dissection (62.5%). A double suction technique was used to control bleeding and haemostasis with a small piece of cotton or a temporary clip, performed in 20 cases (83.3%). However, in cases with premature rupture immediately after the dural or arachnoid opening, the extension of the haematoma into the subarachnoid space resulted in severe brain swelling and partial resection of the brain had to be done to secure temporary clipping. The double suction technique and primary haemostasis using a small piece of cotton or temporary clip resulted in good outcome even in cases with premature rupture. However, very early premature rupture also occurred although its incidence was extremely rare. The removal of part of the brain can secure the working space but the outcome was poor. PMID- 10672296 TI - Complications associated with transvenous embolisation of cavernous dural arteriovenous fistula. AB - Results are presented of transvenous embolisation, via either the inferior petrosal sinus (IPS) or the superior ophthalmic vein (SOV), for 19 patients with cavernous dural arteriovenous fistula with special emphasis on complications. In 17 patients (89%) there was complete angiographic elimination of the shunts and resolution of the symptoms. The remaining two patients also improved clinically, regardless of the minimal residual shunts. Complications included forehead dysaesthesia in one patient, blepharoptosis in two, and transient abducens nerve palsy in three. Injury of the supra-orbital nerve and levator muscle occurred in association with the exposure of the SOV in the patient with dysaesthesia of the forehead and in those with blepharoptosis, respectively. In two patients, abducens nerve palsy resulted from coil over-packing in the cavernous sinus and from dissection of the clival dura during guidewire penetration of the thrombosed IPS in one patient. We found that the complication rate decreased with time, because we became better with this procedure. We believe that transvenous embolisation is the best available treatment modality if one pays careful attention to avoid complications related to the procedure. PMID- 10672297 TI - Does additional discectomy and the degree of dural sac compression influence the outcome of decompressive surgery for lumbar spinal stenosis? AB - The discussion regarding factors that reliably predict the long-term surgical results in patients with lumbar spinal stenosis is still going on. This retrospective study analyses the relation between the dimensions of the dural sac and patients' clinical status before and after decompressive operations performed with or without additional discectomy. The type of surgery performed in 134 patients and the dural sac dimensions measured on postmyelograpic computed tomography in 100 of these patients were related to the Prolo scores before surgery and at follow-up (mean 46 months). The degree of dural sac compression correlated significantly with the patients' postoperative Prolo score and with the difference between the pre- and postoperative scores. The dural sac diameters predicted outcome after surgery more reliably than the preoperative Prolo scores. There was no statistically significant difference in the outcome when comparing patients with and without additional discectomy. The results presented suggest that the relief of symptoms after decompressive surgery for lumbar spinal stenosis correlates with the degree of the dural sac compression and that the simultaneous presence of disc herniation necessitating additional discectomy does not influence the postoperative outcome. However, these results have to be confirmed by prospective studies. PMID- 10672298 TI - Radiosurgery of vestibular schwannomas: a minimally invasive alternative to microsurgery. AB - From April 1992 till December 1998 stereotactic radiosurgery (Gamma Knife) was applied to 192 patients with vestibular schwannomas. 56 of them had radiosurgery as primary treatment modality and were followed-up for at least 4 years (48-80 months, median 62). Without fatal complications, control of tumour growth was achieved in all but three cases, useful hearing being preserved in more than one half of the patients (62%). The neurological state improved in 30 patients (54%). Irradiation-associated adverse effects (18%) comprised neurological signs (incomplete facial palsy, four cases (two recovered completely), and mild trigeminal neuropathy, three cases, respectively) and morphological changes (three patients) marked by an enlargement of pre-existing cystic components calling for additional surgical treatment: Microsurgical decompression was performed in two cases, the third patient underwent a shunting procedure because of hydrocephalus formation. Based on the present data, radiosurgery represents an effective treatment for vestibular schwannomas associated with an exceptionally low mortality rate and a good quality of life. With respect to the preservation of cranial nerve function, results are comparable to microsurgical resection. A short duration of hospitalization and a quick return to normal activities constitute further advantages and contribute to cost effectiveness in public health care. PMID- 10672299 TI - Prolactin secreting pituitary adenomas: analysis of 429 surgically treated patients, effect of adjuvant treatment modalities and review of the literature. AB - OBJECTIVE: We performed this retrospective analysis to determine the efficacy of surgery and radiotherapy over hormonal and volumetric control of prolactinomas, many of which had failed during dopa-agonist therapy. In the same analysis, the efficiency of topical bromocriptine application as a preliminary study was compared with standard treatment modalities. MATERIALS AND METHODS: Between 1982 1997, 429 prolactinoma patients who underwent surgery at Hacettepe University Neurosurgery Department and at Bayindir Medical Center were included in this study. All patients were classified according to Hardy's classification scheme and were further divided into 'invasive' and 'non-invasive' groups based on this radiological classification system. The mean follow-up time was 38.4 months. One hundred and thirty five patients had peroperative bromocriptine application into the sellar cavity and these, either receiving radiotherapy (RT) or not, were analysed separately from the other 294 patients. In the early post-operative period, 104 of these patients were given conventional radiotherapy with median dose of 4500 cGy. We focused on the effects of surgery and radiotherapy over volumetric and hormonal tumour control on the basis of invasion characteristics and the early results of topical bromocriptine application in macroprolactinoma patients; and compared our results with the literature. RESULTS: Statistical analysis revealed that radiotherapy was not effective over hormonal and volumetric tumour control for prolactinomas. We did not observe any correlation to dural invasion of the sellar floor, recurrence, and the disease-free survival time. Topical bromocriptine application seemed to improve the volumetric control in 135 selected macroprolactinoma patients but not hormonal response compared with the standard treatment modalities. CONCLUSION: Conventional radiotherapy is not as effective as expected for prolactinomas and should not be preferred considering its adverse effects. Tumoural infiltration of the sellar dura mater is not a prognostic criterion for recurrence expectation and, therefore, should not be a criterion for radiotherapy after surgery. After subtotal removal, postoperative dopa-agonist therapy should be considered even if the patient was intolerant or resistant to previous treatment since surgery seems to improve patients' drug tolerance and cooperation due probably to the lower dose requirement. The early results of topical bromocriptine application seem to improve volumetric tumour control but this should not be accepted as a judgement since we need to wait for later results and to expand the sample size for more reliable interpretation. PMID- 10672300 TI - Comparison between monopolar and bipolar electrical stimulation of the motor cortex. AB - Intra-operative neurophysiological techniques allow reliable identification of the sensorimotor region and make their anatomical and functional preservation feasible. Monopolar cortical stimulation has recently been described as a new mapping technique. In the present study this method was compared to the "traditional" technique of bipolar stimulation. Functional mapping of the motor cortex was performed in 35 patients during surgery in the central region. The central sulcus (CS) was identified by somatosensory evoked potential (SEP) phase reversal. Cortical motor mapping was first performed by monopolar anodal stimulation with a train of 500 Hz (7-10 pulses) followed by bipolar stimulation (pulses at 60 Hz with max. 4 sec train duration). Surgery was performed under general anaesthesia without muscle relaxants. Of 280 motor responses elicited by bipolar cortical stimulation, 54.23% [152] were located in the primary motor cortex (PMC), 37.85% 106[ outside the motor strip in the secondary motor cortex (SMC), and 8% 22[ posterior to the CS. Of 175 motor responses elicited by monopolar cortical stimulation. 68.57% 120[ were located in the SMC, 23.42% 41[ in the SMC and 8% 14[ posterior to the CS. Contrary to the general clinical view, there is considerable overlapping of primary motor units over a cortical area much broader than the "classical" narrow motor strip along the CS. Bipolar cortical stimulation is more sensitive than monopolar for mapping motor function in the premotor frontal cortex. Both methods are equally sensitive for mapping the primary motor cortex. PMID- 10672301 TI - Nosocomial infections in a neurosurgery intensive care unit. AB - In order to identify overall and site-specific nosocomial infection (NI) rates in patients receiving neurosurgical intensive care therapy, a prospective study was started in February 1997 in the eight-bed neurosurgical ICU of the University Hospital of Freiburg, Germany. Case records were reviewed twice a week, all microbiology reports were reviewed and ward staff was consulted. NI were defined according to the CDC-criteria and were categorised into specific infection sites. Within 20 months, 545 patients with a total of 5,117 patient days were investigated (mean length of stay: 9.4 days). 113 NI were identified in 90 patients (72 pts. with one, 13 with two and 5 with three infections, respectively). A moderate to high overall incidence (20.7/100 pts.) and a moderate incidence density (22.1/1,000 patient days) of NI in the neurosurgical ICU could be documented; these figures are well within the range of published data. Site specific incidence rates and incidence densities were: 1 bloodstream infection per 100 patients (0.9 central line-associated BSIs per 1,000 central line-days), 9 pneumonias per 100 patients (15.1 ventilator-associated pneumonias per 1,000 ventilator-days), 7.3 urinary tract infections per 100 patients (8.5 urinary catheter-associated UTIs per 1,000 urinary catheter-days). Additionally, 1.1 cases of meningitis, 0.7 brain abscesses/ventriculitis, and 1.7 other infections (surgical site infection, bronchitis, catheter related local infection, diarrhoea) were documented per 100 patients, respectively. 14.6% of isolated pathogens were E. coli, 10.2% enterococci, 9.6% S. aureus, 6.4% CNS, 6.4% Klebsiella spp., 5% Enterobacter spp. and 5% Pseudomonas spp. In 11 cases of NI no pathogen could be isolated. PMID- 10672302 TI - DREZ surgery on conus medullaris (after failed implantation of vascular omental graft) for treating chronic pain due to spine (gunshot) injuries. AB - The results of DREZlesioning procedure used for the treatment of chronic intractable pain due to deafferentation caused by gunshot injuries at the thoracolumbar (T10-L1) spine level are reported in six patients. The specificity of these cases arises from the fact that all the patients underwent, after decompressive laminectomy, an implantation of vascularized omental graft on the injured cord segments, 4-17 months after injury. Because of the failure of this method, which did not improve spinal function nor hinder the development of pain, surgery in the DREZ was performed 2-5 years after implantation. The results of the microsurgical DREZotomy procedure in those patients, 7-12 months after the surgery were: 4 patients with complete pain relief and 2 patients with pain relieved of 80%. All the patients with well-confined segmental pain were completely cured. PMID- 10672303 TI - Neurological complications of childhood malignancies. AB - Between Jan 1982 to Jun 1994, 154 children with malignant non-central nervous system tumors, excluding leukemias and lymphomas, were admitted and treated at the UKMC. Fifty-one (33%) of these cases suffered with 64 neurological complications during the course of their diseases. Nine cases suffered with multiple neurological complications. Nervous system metastasis was the most common neurological complication (n = 24; 15.6%), which was followed by nervous system infection (n = 17; 11%). Twelve (7.7%) cases had treatment related peripheral or cranial neuropathies. Seven (4.5%) cases had new onset of grand mall seizures. One case had paraneoplastic syndrome, one case had panhypopituitarism secondary to whole brain radiation, and one case had Horner's syndrome secondary to tumor removal. Ten cases suffered with neurological sequelae secondary to neurological complications. Three of these cases suffered with developmental delay and mental retardation. Fifty-one patients with neurological complications were followed for 9 to 102 months. While 30 (19.7%) patients were alive, 20 (13%) patients died and one case was lost during the analysis of the results. Neuroblastoma/ganglioneuroblastoma has the highest rate for causing neurological complication. IN CONCLUSION: neurological complications were seen on 33% of childhood solid malignant tumors. Nervous system metastasis had the worst prognosis and the most frequent neurological complication. Neurological complications did not increase the mortality rate, but one-third of surviving patients with neurological complications suffered with neurological sequelae. PMID- 10672304 TI - Reversal of cerebral vasospasm by the nitric oxide donor SNAP in an experimental model of subarachnoid haemorrhage. AB - The constant release of nitric oxide (NO) is essential to maintain basal cerebrovascular tone. Oxyhaemoglobin, liberated by lysis of red blood cells after subarachnoid haemorrhage binds NO and prevents its entry into vascular smooth muscle cells. While endothelium-dependent vasoconstriction is preserved, decreased levels of NO inhibit endothelium-dependent relaxation and may cause vasospasm. S-nitrosothiols are potent vasodilators and precursors of NO. The authors' aim was to determine whether S-nitroso-N-acetylpenicillamine (SNAP), a stable S-nitrosothiol compound, could reverse vasospasm in an experimental vasospasm model in rabbit. Experimental subarachnoid haemorrhage (SAH) was induced in 37 New Zealand white rabbits. The animals were divided into four groups. Control (no SAH), SAH only, SAH plus saline and SAH plus SNAP. SNAP (15 micrograms/kg/min) or 0.09% saline (equal volume) was infused 46 hours after induction of SAH. All animals were killed by perfusion fixation 48 hours after SAH occurred. Basilar arteries were removed, sectioned and their cross sectional areas were evaluated in a blind manner, by light microscopy and by using computer assisted morphometry. Experimental SAH elicited vasospasm in all animals of SAH only and SAH plus saline group. In animals treated with SNAP, arterial narrowing was markedly attenuated without producing systemic hypotension. This widening achieved statistical significance when compared to the arteries of the SAH only and SAH plus saline group (p < 0.01). This study indicates that the NO donor SNAP is a potentially useful drug to reverse cerebral vasospasm due to SAH. PMID- 10672305 TI - Experimental study of intracisternal administration of tissue-type plasminogen activator followed by cerebrospinal fluid drainage in the ultra-early stage of subarachnoid haemorrhage. AB - This experimental study evaluated the effect of intrathecal injection of tissue type plasminogen activator followed by cisternal drainage in the ultra-early stage of aneurysmal subarachnoid haemorrhage to prevent vasospasm. Twenty Japanese white rabbits were divided into five groups. Either tPA (groups A, B, and E) or saline (groups C and D) was injected intrathecally 1 hour (groups A, B, C, and D) or 21 hours (group E) after the intrathecal injection of blood. Cerebrospinal fluid was drained 2, 4, and 6 hours after the intrathecal injection of blood (groups A, C, and E). On day 4, the angiographic caliber of the basilar artery in each group was as follows (mean +/- SD): A, 85.9 +/- 5.0%; B, 74.6 +/- 5.3%; C, 69.1 +/- 2.7%; D, 64.0 +/- 4.9%; E, 80.2 +/- 2.7% (compared with baseline). In the two groups in which CSF was drained (groups A and C), fibrinolysis with tPA significantly suppressed vasospasm. In the two groups treated with tPA (groups A and B), cisternal drainage significantly suppressed vasospasm. In the two groups treated with saline (groups C and D), however, cisternal drainage did not suppress vasospasm. Examination of the series of CSF samples (groups A and C) showed that fibrinolysis with tPA effectively cleared clots early. In the two groups treated with tPA and CSF drainage (groups A and E), early removal of subarachnoid clots reduced the degree of vasospasm. Early fibrinolysis with tPA and early removal of subarachnoid clots by drainage is effective for preventing vasospasm. PMID- 10672306 TI - Ganglioside profiles in human gliomas: quantification by microbore high performance liquid chromatography and correlation to histomorphology and grading. AB - The composition and the content of gangliosides changes during physiological growth and differentiation as well as in neoplastic cell transformation. In order to determine if ganglioside profiles correlate with brain tumour malignancy, the ganglioside distribution was determined in 31 gliomas of astrocytic origin and in non-tumour tissue by a recently developed microbore high performance liquid chromatography (HPLC) method. Glioma malignancy was graded according to the grading system proposed by the World Health Organization (WHO) in 1993. In general, an increase of GD3 and a decrease of normal brain gangliosides correlated with a higher grade of malignancy. Pilocytic astrocytomas Grade I had a distinctive ganglioside profile, histologically as well as biochemically. Although they are low-grade gliomas, the pilocytic astrocytomas exhibited a GD3 content comparable to anaplastic gliomas and could only be biochemically distinguished from other tumour grades by relatively high type "b" ganglioside levels. Thus, ganglioside composition not only reflects anaplasia but can also be used to indicate biological characteristics of tumours of different histogenetic origin. PMID- 10672307 TI - Cerebellopontine angle lipoma: case report and review of the literature. AB - Intracranial lipomas located in the cerebellopontine angle are extremely rare. These tumours are mal-developmental lesions which can cause slowly progressive neurological symptoms. The clinical management of these tumours differs significantly from other lesions in this region. A 27 year old woman presented with a 2-month history of vertigo and a slowly progressive deterioration of hearing in the left ear. Computed tomography (CT) revealed a large low-density mass in the left cerebellopontine angle without any contrast-enhancement. In T1 weighted magnetic resonance imaging (MRI) the lesion was hyper-intense and did not enhance after application of gadolinium. Areas of lower signal intensity inside of the lesion were suggested as incorporated cranial nerves. A left retro sigmoidal approach in a semi-sitting position was chosen to expose the tumour. After reducing the tumour mass, the tumour was dissected from the cranial nerves which were incorporated into the tumour. The residual tumour was adherent to the brain stem and the encased lower cranial nerves, allowing only a near subtotal resection of the highly vascularized tumour in order to avoid neurological deficits. The histological examination revealed a lipoma. Attempts at complete removal of cerebellopontine angle lipomas usually result in severe neurological deficits. Conservative treatment should therefore be preferred. Limited surgery is indicated if the patients suffer from disabling neurological symptoms and signs e.g., vertigo, nausea, trigeminal neuralgia, facial weakness or facial spasm. PMID- 10672308 TI - Replantation of a totally avulsed scalp without microvascular anastomosis. PMID- 10672309 TI - Malignant non-Hodgkin lymphoma presenting as a nerve root tumour. PMID- 10672310 TI - Simultaneous transcranial and endoscopic transnasal approach for recurrent huge pituitary adenoma. PMID- 10672311 TI - Transient mutism after brain stem infarction. PMID- 10672312 TI - Referring to the posterior fossa cranioectomy and tonsillar resection in order to treat Chiari I malformation with syringomyelia. PMID- 10672313 TI - Selective distribution of matrix metalloproteinase-3 (MMP-3) in Alzheimer's disease brain. AB - A growing amount of evidence indicates that matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of Alzheimer's disease (AD). Stromelysin-1 (MMP-3) plays a central role in activating latent-type MMPs, which are originally secreted as proenzymes. We examined MMP-3 immunoreactivity in the brains of patients who had suffered from Alzheimer's disease and in those of neurologically normal persons. The interstitium between myelinated axons and astrocytes in the white matter of all brain tissues, and senile plaques in the gray matter of the patients with AD were stained with a monoclonal antibody to MMP-3. Comparison of the number of senile plaques stained with the antibody against MMP-3 in the parietal cortex with that in the hippocampus showed that fewer plaques were stained in the hippocampus. The selective distribution of MMP 3 in the human brain suggests that MMP-3 might play an important role in the pathogenesis of AD, especially in the degradation of beta-amyloid protein. PMID- 10672314 TI - Differential expressions of protein kinase C isozymes during proliferation and differentiation of human skeletal muscle cells in vitro. AB - The mechanism of skeletal muscle regeneration in vivo can be well modeled in vitro by culturing skeletal muscle cells. In these cultures mononuclear satellite cells fuse to form polynuclear myotubes by proliferation and differentiation. The aim of this study was to determine how the different protein kinase C (PKC) isozymes were expressed during differentiation of human skeletal muscle in vitro. The expressions of desmin, used as a muscle-specific intermediate filament protein marker of differentiation, and of different PKC isozymes were detected by single and double immunohistochemical labeling, and by Western blot analysis. In skeletal muscle cells we could identify five PKC isozymes (PKC alpha, -gamma, etha, -theta and -zeta). The expressions of PKC alpha and -zeta did not change significantly during differentiation; their levels of expression were high in the early immature cells and remained unchanged in later phases. In contrast, the expression levels of PKC gamma and -etha increased with differentiation. Furthermore, the cellular localization of PKC gamma markedly altered during differentiation, with a perinuclear-nuclear to cytoplasmic translocation. The change in the level of expression of PKC theta during differentiation showed different pattern; its expression was high during the early phases, but a decreased immunostaining was detected in the matured, well-differentiated myotubes. We conclude, therefore, that cultured human skeletal muscle cells possess a characteristic PKC isozyme pattern, and that the different phases of differentiation are accompanied by different expression patterns of the various isozymes. These data suggest the possible functional and differential roles of PKC isozymes in human skeletal muscle differentiation. PMID- 10672315 TI - Morphological alterations of the choroid plexus in late-onset Alzheimer's disease. AB - Anomalies of the cerebrospinal fluid flow rate and composition that have been reported in patients suffering from Alzheimer's disease (AD) could be related to alterations of the choroid plexuses (CD). Here we report a photonic and electron morphometric study in which we compared the height of CP epithelial cells and the thickness of their basement membrane on post-mortem samples from AD patients, age matched controls and two newborns. Ageing appeared associated with epithelial atrophy and basement membrane thickening, but these features were significantly accentuated in AD. These data suggest that a dramatic alteration of the secretion and filtration could be involved in the multiparametric pathogenesis of late onset AD. PMID- 10672316 TI - Time course of ultrastructural changes and immunoelectron microscopic localization of neurocalcin in motor endplates of the lumbrical muscles of rats given a single administration of 2,5-di(tert-butyl)-1,4-hydroquinone. AB - A time-course study of ultrastructural changes and immunoelectron microscopic localization of neurocalcin was performed on motor endplates of the lumbrical muscles of female Wistar rats given a single oral administration of 2,5-di(tert butyl)-1,4-hydroquinone (DTBHQ) at a dose of 120 mg/kg. Toxic signs such as salivation and muscle weakness of the hind legs appeared from 3 h after DTBHQ administration. No remarkable macroscopic or light microscopic changes were noted in the lumbrical muscles of the treated rats. At the ultrastructural level, neurotoxicity characterized by a decreases or loss of synaptic vesicles and mitochondria was observed after 24 h and at the 1-week time point, nerve endings had disappeared in some of the motor endplates, while many neurite nerve endings suggestive of early stage regeneration were apparent. After 6 weeks, newly formed reinnervated endplates were observed. Immunoelectron microscopically, the synaptic vesicle membranes were heavily labeled for neurocalcin in the control rats, but not at 24 h after DTBHQ treatment. Synaptic vesicle membranes in the DTBHQ group were weakly labeled at 1 week, but strongly at 6 weeks. The results strongly suggest that DTBHQ targets the motor endplates in the rat lumbrical muscles, causing depletion of neurocalcin in the synaptic vesicles followed by their loss. PMID- 10672317 TI - Tissue tears in the white matter after lateral fluid percussion brain injury in the rat: relevance to human brain injury. AB - A characteristic feature of severe diffuse axonal injury in man is radiological evidence of the "shearing injury triad" represented by lesions, sometimes haemorrhagic, in the corpus callosum, deep white matter and the rostral brain stem. With the exception of studies carried out on the non-human primate, such lesions have not been replicated to date in the multiple and diverse rodent laboratory models of traumatic brain injury. The present report describes tissue tears in the white matter, particularly in the fimbria of Sprague-Dawley rats killed 12, 24, and 48 h and 7 days after lateral fluid percussion brain injury of moderate severity (2.1-2.4 atm). The lesions were most easily seen at 24 h when they appeared as foci of tissue rarefaction in which there were a few polymorphonuclear leucocytes. At the margins of these lesions, large amounts of accumulated amyloid precursor protein (APP) were found in axonal swellings and bulbs. By 1 week post-injury, there was macrophage infiltration with marked astrocytosis and early scar formation. This lesion is considered to be due to severe deformation of white matter and this is the first time that it has been identified reproducibly in a rodent model of head injury under controlled conditions. PMID- 10672318 TI - New haplotype of familial Creutzfeldt-Jakob disease with a codon 200 mutation and a codon 219 polymorphism of the prion protein gene in a Japanese family. AB - We report a new haplotype of familial Creutzfeldt-Jakob disease (CJD) with a codon 200 mutation and a codon 219 polymorphism of the prion protein gene in a Japanese family. There were four cases diagnosed with CJD neuropathologically, one of which was identified with a codon 200 mutation (glutamic acid to lysine) and a codon 219Lys polymorphism on the same allele. Clinicopathologically, two cases had a long clinical course, whereas the others were similar to the cases with a codon 200 mutation. Three cases was diagnosed with the panencephalopathic type CJD neuropathologically and the other was diagnosed with the subacute spongiform encephalopathy, a subtype of CJD. We consider that the clinicopathological features in familial CJD are not steadily uniform and that it is impossible to state definitely from this study whether the codon 219 polymorphism influences the clinicopathological aspects in familial CJD with a codon 200 mutation (glutamic acid to lysine). PMID- 10672319 TI - Glial cell line-derived neurotrophic factor (GDNF) and its receptor (GFR-alpha 1) are strongly expressed in human gliomas. AB - Glial cell line-derived neurotrophic factor (GDNF), a sequence-related factor of the transforming growth factor-beta family, has been identified as a potent neurotrophic factor for a variety of neuronal cell populations. At present, it is still unknown whether human gliomas in vivo are also capable of producing GDNF. We studied the expression of GDNF in 14 human glioblastomas, 1 gliosarcoma and 5 astrocytomas. Using an enzyme-linked immunosorbent assay, the amount of GDNF was quantified in human gliomas and compared to GDNF-expression in C6 glioma cells, mouse fibroblasts and normal human and rat brain. Mean concentration of GDNF in gliomas was 937 +/- 140 pg GDNF/g tissue (n = 20). C6 cells revealed the highest expression levels of 2,837 +/- 813 pg/g, whereas mouse 3T3 fibroblasts showed no detectable GDNF protein. Mean GDNF tissue levels in normal human and rat brain were significantly lower. Using reverse transcriptase-polymerase chain reaction, GDNF mRNA was detected in human gliomas and in rat C6 cells. Immunohistochemistry revealed strong GDNF- and GDNF receptor-alpha 1-expressing tumor cells in human glioma tissue. These results show that glial tumors, even in the most dedifferentiated form of glioblastoma, express GDNF at concentrations up to five times higher compared to normal human brain. This overexpression of GDNF may be of biological relevance for proliferation of glial tumors in humans. PMID- 10672320 TI - Up-regulation of the hyaluronate receptor CD44 in canine distemper demyelinated plaques. AB - CD44 antigen (CD44), the principle cell surface receptor for hyaluronate, is up regulated in the human demyelinating disease multiple sclerosis on fibrous astrocytes. As astrocytes are the main target cell of canine distemper virus (CDV), the consequences of a CDV infection on the CD44 expression and distribution in brains with spontaneous demyelinating canine distemper encephalitis (CDE) were of interest. Thirteen acute, 35 subacute, and 11 chronic plaques of nine dogs with immunohistologically confirmed CDE and brains of control dogs were included in the study. For light microscopy, 5-micron-thick serial sections were stained with H&E and incubated with monoclonal antibodies (mAbs) against CD44 and canine distemper virus nucleoprotein and polyclonal antibodies (pAbs) against glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP). For immunoelectron microscopy, 90-nm-thick sections were double stained with anti-GFAP and anti-CD44 mAbs to specify CD44-expressing structures. In controls, CD44 was diffusely distributed in the white matter and single meningeal cells exhibited a marginal expression of the antigen. In acute and more prominently in subacute demyelinating encephalitis, there was a plaque-associated up-regulation of CD44 which paralleled GFAP. In chronic demyelinating lesions, a reduction of CD44 associated with a loss of GFAP-positive astrocytes was noted. Additionally, in chronic plaques, CD44 was expressed on the cell membrane of perivascular mononuclear cells. Immunoelectron microscopically, in controls, CD44 was rarely demonstrated on astrocytic cell processes. In contrast, in brains with CDE CD44 was found on the cell membrane of broadened astrocytic cell processes. In summary, CD44 is up-regulated on astrocytes in the early phase of CDE and seems to represent a marker for the activation of immune cells in the late phase of the infection. PMID- 10672321 TI - Microglial and astrocytic reactions prior to onset of thalamic cell death after traumatic lesion of the rat sensorimotor cortex. AB - The temporospatial relationship between microglial and astrocytic reactions and delayed thalamic cell death was examined 1-7 days following a traumatic cold lesion of the rat sensorimotor cortex using immunocytochemistry in combination with terminal deoxynucleotidyltransferase-mediated biotinylated dUTP nick end labeling (TUNEL) of nuclear DNA fragmentation. No or only occasional TUNEL positive cells were found in the thalamic relay nuclei up to 3 days after trauma. After 7 days, on the other hand, a considerable number of TUNEL-positive cells were seen in the ventrobasal, the ventrolateral and posterior thalamic nuclei. Already 3 days after trauma, i.e., before cell injury was detectable, many protoplasmic astrocytes, which were reactive for glial fibrillary acidic protein, and ramified microglia, which were positive for complement receptor type 3b (CR3b) but negative for major histocompatibility complex (MHC) class II antigen, were noticed in the thalamus. The number of labeled astro- and microglia further increased after 7 days, when DNA fragmentation became evident. At this time, the morphology of microglia shifted towards bushy and rod-like cells, and microglia became also reactive for MHC class II antigen. Clusters of CR3b- and MHC class II positive microglia were found in the ventrobasal thalamus. The present findings demonstrate that trauma-induced microglial and astrocytic reactions appear in the thalamus prior the onset of cell damage. PMID- 10672322 TI - alpha-Synuclein is expressed in a variety of brain tumors showing neuronal differentiation. AB - alpha-Synuclein is presynaptic nerve terminal protein and its immunoreactivity has been observed in such neurodegenerative structures as senile plaques of Alzheimer's disease or Lewy bodies of Parkinson's disease. The physiological role of alpha-synuclein is still unknown. It is speculated that alpha-synuclein may be expressed in brain tumors, especially in those showing neuronal differentiation. We examined the immunohistochemical localization of alpha-synuclein in 77 human brain tumors. alpha-Synuclein was widely distributed in the brain tumors showing neuronal differentiation. As a result, positive immunostaining for alpha synuclein was observed in ganglioglioma, medulloblastoma, neuroblastoma, primitive neuroectodermal tumor, pineocytoma/pineoblastoma, and central neurocytoma. Compared with other neuronal markers, the positive ratio of alpha synuclein was not as high as synaptophysin, microtubule-associated protein 2, neuron-specific enolase and tau, but it was higher than neurofilament and chromogranin A. The expression of synaptophysin was diffusely observed in the cytoplasm, cellular processes and nucleus in tumors showing neuronal differentiation; however, the expression of alpha-synuclein was predominantly observed in the cytoplasm of the tumors as well as in the cellular processes. On the other hand, non-neuronal brain tumors such as astrocytic tumors or meningiomas were totally negative for alpha-synuclein. In conclusion, the appearance of an alpha-synuclein-positive structure was not limited to neurodegenerative diseases, but could also be detected in neoplastic cells showing neuronal differentiation. PMID- 10672323 TI - Distribution of enzyme-bearing cells in GM2 gangliosidosis mice: regionally specific pattern of cellular infiltration following bone marrow transplantation. AB - Tissue distribution of beta-hexosaminidase was investigated using 5-bromo-4 chloro-3-indolyl N-acetyl beta-D-glucosaminide (X-Hex) as substrate in wild-type mice, four GM2 gangliosidosis model mice (Hexa-/-, Hexb-/-, Gm2a-/- and Hexa-/ Hexb-/-) and Hexb-/- mice that received bone marrow transplantation (BMT). In wild-type mice histochemical localization of beta-hexosaminidase was detected in the perikarya of the majority of neurons, small process-bearing microglial cells, perivascular macrophages, and macrophages in the choroid plexus and leptomeninges. X-Hex positivity was also noted in the renal tubular epithelium and macrophages in the liver and spleen. The staining pattern in the Gm2a-/- and Hexa-/- mice was generally similar to those of wild type, but in these mice, X Hex stain was also noted in some storage neurons with swollen perikarya. No X-Hex positive cells were detected in Hexb-/- or Hexa-/-Hexb-/- (DKO) mice. In Hexb-/- mice that received wild-type BMT (Hexb-/- +WBMT), many X-Hex-positive cells were detected in the spleen, and to a far lesser extent, in liver and kidney. In the CNS of these mice, X-Hex-positive cells were largely detected in the leptomeninges and choroid plexus. Some positive cells were also detected, mostly in the perivascular regions of the cerebrum, in particular in the regions of the posterior thalamus, brain stem and spinal cord. Some of X-Hex-positive cells were immunoreactive with Mac-1 and F4/80 antibodies and, thus, were cells of microglia/macrophage lineage. X-Hex-positive staining was not detected in neurons in these mice despite clinical improvement following BMT. This is the first time, as far as we know, that the regional distribution of the donor cells in the CNS has been investigated in a model of neuronal storage disease. Our study indicated that donor-derived cells of microglia/macrophage lineage infiltrated the CNS in a regionally specific manner following the BMT. PMID- 10672324 TI - Postnatal profiles of myogenic regulatory factors and the receptors of TGF-beta 2, LIF and IGF-I in the gastrocnemius and rectus femoris muscles of dy mouse. AB - The dystrophin-deficient mdx mouse presents muscle fiber necrosis but active muscle regeneration, probably due to an extensive recruitment of myogenic regulatory factors (MRF), several growth factors and cytokines, and favorable interaction of satellite cells. In contrast, the laminin alpha 2 (merosin) deficient dy mouse shows progressive muscle fiber necrosis and ineffective muscle regeneration. Using Western blot and immunohistochemical analyses, we investigated the adaptive changes in MRF, growth factors and cytokines and their receptors in the muscles of dy mice during postnatal growth. The relative volume of MyoD, myogenin and Myf-5 proteins was markedly lower in the gastrocnemius and rectus femoris muscles of dy mice. Transforming growth factor-beta 2, leukemia inhibitory factor (LIF) and basic fibroblast growth factor were not up-regulated in the muscles of dy mice. The levels of the LIF receptor and insulin-like growth factor-I receptor levels were markedly decreased in the muscles of dy mice during the entire postnatal period observed in this study. Therefore, unlike the situation in mdx mice, the milieu of regeneration following repetitive damage seems to be degraded in the muscles of dy mice. PMID- 10672325 TI - The adaptive response of transforming growth factor-beta 2 and -beta RII in the overloaded, regenerating and denervated muscles of rats. AB - Using a muscle cell line and satellite cell cultures, it has been shown that transforming growth factor-beta (TGF-beta) has a powerful inhibitory effect on myoblast replication and differentiation. However, little work has been done on the possible role of TGF-beta in adult muscle in vivo. Using Western blot and immunohistochemical analyses, we investigated normal distribution of TGF-beta 2 and TGF-beta RII proteins between slow and fast-type muscles, and the adaptive response of these proteins in the mechanically overloaded muscles, in the regenerating muscles following bupivacaine injection and in the denervated muscle after section of sciatic nerve. Slight TGF-beta 2 immunoreactivity was detected both in slow- and fast-type muscles of mature rat. The amount of TGF-beta RII protein was markedly greater in fast-type muscles. In the overloaded muscle, immunohistochemical analysis showed a marked increase in TGF-beta 2 immunoreactivity in the mononuclear cells (probably endothelial and perithelial or smooth muscle cells of endomysial capillaries) of the extracellular space at 3 and 6 days post surgery. Rapid increase of TGF-beta 2 protein and concomitant decrease of the receptor (TGF-beta RII) were observed in the mechanically overloaded and regenerating muscles. On the other hand, denervation of slow- and fast-type muscles showed a rapid increase in TGF-beta 2 protein, but did not elicit a concomitant decrease of TGF-beta RII. These results indicate that TGF beta RII is preferentially distributed in fast-type muscles. Furthermore, TGF beta 2 may play an important role in muscle hypertrophy and regeneration by the usage of TGF-beta RII. PMID- 10672326 TI - Single-fiber analysis of mitochondrial A3243G mutation in four different phenotypes. AB - Five unrelated patients harboring the A3243G mutation in the mitochondrial DNA (mtDNA) but presenting with different clinical phenotype were studied for their percentage of mutation at the single muscle fiber levels. One patient had a clinically and pathologically defined Leigh syndrome (LS), two showed mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), another showed progressive external ophthalmoplegia (PEO), and the other showed mitochondrial diabetes mellitus (MDM). The mutation load was greater in the muscle from the patient with LS (92%), who showed more than 80% even in the non-ragged red fibers (RRF) and also presented the highest proportion of RRF. The patients with MELAS had lower mutation levels as well as a lower proportion of RRF, and these two parameters were even lower in the PEO and MDM patients. These results were consistent with the concept that differences in the mutation load and in the somatic distribution of the mutation among different cells and tissues are responsible for the differences in phenotypical expression of the disease. PMID- 10672327 TI - Expression of p67 (Munc-18) in adult human brain and neuroectodermal tumors of human central nervous system. AB - p67 (Munc-18), is a neuron-specific protein of 67 kDa, known for its ability to bind with syntaxin and also to copurify with neuronal cdc2-like kinase. Earlier, in situ hybridization and immunocytochemical analysis of rat trigeminal ganglion and hippocampal cells demonstrated the specific localization of p67 in nerve cells and its rich distribution in axons. In the present study, we have looked for p67 expression in normal human brain and various neuroectodermal tumors. Immunohistochemical and Western immunoblot analysis of normal human brain tissue using antibodies against the N- and C-termini of p67 demonstrated the specific localization of this protein in postmitotic neurons but not in glia. Among neuroectodermal tumors, expression of p67 was observed in 100% of the tumors of neuronal origin studied, especially in the mature neuronal cell population of these tumors. Western immunoblot analysis of non-neuronal neuroectodermal tumors failed to reveal the expression of this protein in majority of cases. However, in gliomas and meningiomas, mild cytoplasmic immunohistochemical staining of neoplastic cells was noted in 64.7% and 25% of cases, respectively. Observed mild immunohistochemical staining of these tumors could be due to immunoreactivity to low molecular weight degraded products of p67, as seen on Western blot. The findings suggest that p67, by virtue of its ability to be expressed in postmitotic neurons of adult human brain and in tumors of neuronal origin, may serve as a molecular tool to understand the growth and differentiation of the nervous system in general. PMID- 10672328 TI - Practical measures to simplify the Braak tangle staging method for routine pathological screening. AB - The examination of neurofibrillary tangles is now recommended for the diagnosis of Alzheimer's disease as their location and density can distinguish early, intermediate and late disease stages. While the Braak tangle staging protocol can identify these stages, it uses an uncommon silver stain and hippocampal sample. The present study evaluates the Braak protocol using commonly used methods and cases fulfilling either CERAD criteria for Alzheimer's disease, criteria for dementia with Lewy bodies or without neurological disease. Temporal and occipital cortices from 72 cases were stained using tau immunohistochemistry and the Gallyas and modified Bielschowsky silver stains. The modified Bielschowsky silver stain was equivalent to the Gallyas silver stain for tangle staging. Semiquantitative evaluation of neurofibrillary tangles in the hippocampus and the inferior temporal cortex provided equivalent information to that obtained using the original Braak tangle staging protocol (kappa statistic of 0.97). Comparison of this modification with the CERAD criteria provided moderate agreement (0.51) between diagnostic categories when cases with dementia with Lewy bodies were included, but substantially increased agreement (0.74) when they were excluded. This simplification of the Braak tangle staging protocol is easy to apply, can be readily incorporated into existing CERAD procedures, and helps to distinguish cases with neurofibrillary tangles from those with Lewy bodies. PMID- 10672329 TI - The cerebellar and thalamic degeneration in Fukuyama-type congenital muscular dystrophy. AB - We report a male autopsy case of Fukuyama-type congenital muscular dystrophy (FCMD), with unusual neuropathological findings. The patient was a Japanese man aged 26 years at the time of death. He had shown severe psychomotor retardation and muscular dystrophy since early infancy, and was diagnosed as having FCMD at the age of 5 years. He died of respiratory failure. The main neuropathological finding was extensive cerebral and cerebellar cortical dysplasia, characteristic of this disorder. In addition, degeneration of the cerebellar efferent pathway, including the dentate nucleus, superior cerebellar peduncle, and red nucleus, and that of the lateral thalamic nucleus were observed. These findings suggest the possibility that the long survival can clarify the latent neurodegeneration in the cerebellum and thalamus in FCMD, in addition to congenital malformations. The system degeneration should be carefully evaluated in the pathological examination of this disorder. PMID- 10672330 TI - Eosinophilic inclusions in ependymoma represent microlumina: a light and electron microscopic study. AB - A study was undertaken to determine the pathological significance of previously unrecognized intracytoplasmic eosinophilic inclusions (IEIs) in ependymoma. The study group consisted of 58 ependymomas, all of which were pathologically characterized and graded according to the 1993 WHO classification. Electron microscopic studies were performed in 16 cases. The study showed that 33 (57%) ependymomas had IEIs and that in 8 cases these were abundant. Round and eosinophilic, their sizes varied from 10 microns to a tiny dot. Similar eosinophilic bodies were also observed between tumor cells. The inclusions were weakly PAS positive. On immunostains, IEIs were frequently positive for glial fibrillary acidic protein, less often for S-100 protein, and for epithelial membrane protein and CAM 5.2. They were negative for AE1/AE3, carcinoembryonic antigen and Ber-EP4. Ultrastructurally, IEIs represented intracytoplasmic lumens containing microvilli and cilia. These microlumina also frequently contained granulo-tubular materials. With reference to tumor subtypes, IEIs occurred most frequently in ordinary and clear cell ependymomas. IEIs were also present in 4 of 6 anaplastic ependymomas studied. In conclusion, IEIs represent microlumina and occur in more than a half of ependymomas including malignant examples. Their finding is a helpful diagnostic feature of ependymoma. PMID- 10672331 TI - Histological and immunocytochemical data on the differentiation of intestinal endocrine cells in human fetus. AB - The differentiation of the argyrophil, argentaffin, 5-hydroxytryptamine (5-HT)-, somatostatin (SOM)-, cholecystokinin (CCK)-, substance P (SP)-, methionine enkephalin (Met-Enk)- and vasoactive intestinal polypeptide (VIP)-immunoreactive entero-endocrine cells (EECs) was examined in human fetuses. A great increase in the frequency of EECs in the duodenum and the rectum was observed between the 7th and 12th gestation week. The differentiation of the EECs advanced distally in the small intestine and proximally in the large intestine. In 24-25-week-old fetuses the frequency of the EECs was also increased in the ileum and the colon. A different time-course of the appearance and differentiation of the types of EECs was observed. Met-Enk- and VIP-immunoreactive endocrine cells were not detected at any age. A regional difference in the frequency and morphology of the endocrine cell types was observed in the eldest fetuses. PMID- 10672332 TI - Renal excretory function in conscious Long Evans and vasopressin deficient (Brattleboro) rats after endothelin-A receptor inhibition. AB - All experiments were performed on conscious, freely moving male Long Evans as well as Diabetes incipidus (Brattleboro) rats (300-320 g). The endothelin-A (ETA) receptor antagonist BQ-123 (Neosystem) was administered through femoral vein cannula. Arterial blood pressure was measured trough femoral artery catheter. The bladder was cannulated for urine collection via a small suprapubic incision. After a 40 min control period BQ-123 infusion (16.4 nmol/kg/min, 25 microliters/min) was started and continued for 50 min. The effect of 32.8 nmol/kg/min BQ-123 infused in conscious Brattleboro rats was also investigated. Plasma and urine concentrations of sodium, potassium and chloride as well as osmolality were determined. Glomerular filtration rate (GFR) was estimated using the clearance of endogenous creatinine. Endothelin-A receptor inhibition by 16.4 nmol/kg/min BQ-123 infusion in conscious Long-Evans rats decreased urine flow rate by 38.4% (p < 0.02) and increased urine osmolality by 30.3% (p < 0.05). Sodium, potassium, chloride excretion did not alter. Endothelin-A receptor inhibition by 16.4 nmol/kg/min and by 32.8 nmol/kg/min BQ-123 infusion in conscious Brattleboro rats did not produce any change in urine flow rate, urine osmolality or excretion of the electrolytes studied. Endothelins acting via ETA receptors may function as an inhibitor of water reabsorption in the kidneys of conscious rats. PMID- 10672333 TI - Spectral analysis of heart rate and arterial pressure variability after nitric oxide synthase inhibition. AB - The experiments were performed on male, conscious Wistar rats. Femoral arterial pressure was registered by Statham GOULD P23 ID pressure transducer connected to MP 100WS BIOPAC work station after analog to digital conversion during 40 minutes long control period. Nitric oxide synthase inhibition was performed by injection of 100 microliters, 10 mg/kg b.w. N-omega-nitro-L-arginine methyl ester (L-NAME) in saline through femoral vein catheter. Twenty minutes later arterial pressure registration was started and was continued for 40 minutes. The pulse-by-pulse values of systolic, diastolic and mean arterial pressure as well as the pulse intervals were measured by peak and rate detectors of the AcqKnowledge 2.0 software. Row data were processed using a virtual instrument developed in our laboratory in the graphical programming environment Lab VIEW 3.1.1. L-NAME increased systolic, diastolic and mean arterial pressure by 16.6%, 25% and 35%, respectively. The PMF/PHF ratio in heart rate spectrum decreased, indicating an increased vagal effect on the heart. Nitric oxide synthase inhibition increased the low-frequency component of systolic arterial blood pressure variability by 39.5%. Nitric oxide is a physiological regulator of rapid fluctuations of arterial blood pressure. PMID- 10672334 TI - Neuropeptides of the cholecystokinin group: effects and mechanisms of action on the gastro-intestinal and gall bladder motility. AB - The neuropeptides of the cholecystokinin (CCK) group belong to the substances usually referred to as "brain-gut" neuropeptides. They are synthesized in neurons of the central nervous system, in the peripheral and in the autonomous nervous systems, in endocrine cells (types "I", "K" and "A"), as well as in the enteric nervous system of the gastro-intestinal tract and of the pancreas. The CCK-group peptides realize their effects via several different mechanisms (Fig. 1): endocrine or neuroendocrine (classic hormonal mechanism)--the peptide, released by the endocrine cell or by the nerve terminal, is carried by the circulation to the remote target organs; paracrine or neuroparacrine--the peptide, released in the intercellular space, reaches the target effector cells via diffusion. Similarly to the classic neurotransmitters, CCK and its analogues could play a neurotransmitter role, also modulating the release of acetylcholine (ACh) and of other neurotransmitters in enteric and CNS neurons. In the present review article some smooth-muscle and neuromodulatory effects of CCK are described and compared to the results of the authors' studies on the problem. PMID- 10672335 TI - Does nitric oxide participate in the mechanism of action of dotarizine? AB - Behavioral and nociceptive effects of dotarizine (DOT) and other substances acting on migrainous attacks and nitric oxide (NO) metabolism were studied in comparative experiments on rats. Behavioral effects were evaluated by the changes induced in ambulations and rearings of rats in the Opto-Varimex apparatus; effects on nociception were determined by the changes of pain threshold in growing mechanical pressure on one of the rat paw. The data showed that (1) NO did not participate directly in the mechanism of the behavioral actions of DOT. A role could be ascribed to the modulating influence of DOT on the changes in NO formation induced by other agents; (2) the NO system did not participate in the mechanisms of the responses to the painful mechanical pressure on the rat paw; (3) the behavioral effects of the substances with facilitating or inhibitory action on the migrainous process (m-CPP and ergotamine) and the influence of substances proved to affect NO formation (L-arginine, histamine, L-NAME) on these effects suggest a role for NO as a modulating but not a basic factor in the mechanisms of action of these pro- and antimigrainous substances; and (4) the behavioral effects of DOT were similar to the effects of the antimigrainous drug ergotamine and different from the promigrainous drug meta-chlorophenyl-piperazine (m-CPP)--which suggest an antimigrainous activity of dotarizine. PMID- 10672336 TI - An apparatus for measuring the optical density of human ocular media to blue light. AB - The paper describes an apparatus constructed by the authors and aimed at evaluating the optical density of human ocular media by a non-invasive psychophysical method described previously (Sample et al. 1988). The device has two light sources within the wavelength bands of 437 +/- 6 nm and 557 +/- 6 nm, respectively. They are a square-wave alternated at 1 Hz. The radiance of each can be controlled by the subject and adjusted at visibility threshold or at subjective equiluminance. Upon scotopic vision, the ratio between the two thresholds (or ratio of radiances at equiluminance) is a function of the ocular optical density at the short wavelength. The psychophysical procedures of adjustment of the 437 nm source at the absolute threshold and at equality with the 557 nm source had been tested with volunteers aged between 24 and 67 years. The dependence of blue-light sensitivities on age, obtained with the two procedures, suggests, in agreement with the literature, two main sources of sensitivity decline with age: senile myosis and ocular-media density increase at the short wavelength end of the visible spectrum. PMID- 10672337 TI - Frog muscle fibre action potential and different extracellular calcium concentration at lowered pH in the medium. AB - This article is mainly concerned with the influence of Ca2+[o in acidified extracellular medium on the intracellular action potentials (ICAPs) and total ionic current (Ii) during ICAP of isolated skeletal muscle fibre. The bundles of frog muscle fibres were bathed in Ringer's solution with standard Ca2+[o at pH 6.5 after which the fibres were exposed for 30 min to Ca(2+)-free solution and Ca(2+)-enriched solution at pH 6.5. The ICAPs in standard Ca2+[o solution (control) and after exposure for 30 min to Ringer's solutions with different Ca2+[o at pH 6.5 were recorded and the Ii during ICAP was calculated. The ICAP amplitude from the fibres in Ca(2+)-free solutions at pH 6.5 showed a significant increase vs. control, while the time characteristics if the ICAPs in different Ca2+[o decreased except for the ICAP depolarization phase duration in Ca(2+) enriched solution. The Ii alterations reflect ICAP changes. It was suggested that the changed Ca2+[o at pH 6.5 compensated to some extent the observed inhibitory effect of lowered pH on ICAP parameters in solution with standard Ca2+[o. PMID- 10672338 TI - The "small" polydisperse cytoplasmic extrachromosomal DNA of chicken leukaemic myeloblasts and the avian myeloblastosis virus core-bound DNA seem to descend from origin regions of chromosomal DNA replication. AB - Nucleotide sequences are presented for 12, 7 and 12 cloned extrachromosomal DNAs by nature harbored in nucleoprotein (NP) complexes forming chicken leukaemic myeloblast (CHLM) post microsomal sediment (POMS) components A, B and C, respectively, and for 11 cloned avian myeloblastosis virus (AMV) DNAs. Analysis of the abundance of sequence motifs significant for eukaryotic chromosomal DNA replication origin (ori) regions (and their initiation zones) has shown that these DNAs are reminiscent of cell DNA fragments enriched in ori sequences (Rao et al., 1990) and/or sequence features of several eukaryotic chromosomal oris containing clusters of modular sequence elements (Dobbs et al., 1994). Accordingly, these DNAs, with an (A + T) content prevalently higher than that of the total cell DNA, revealed the presence of asymmetrically distributed (A + T) rich stretches, scaffold attachment region (SAR) T consensuses, polypyrimidine nucleotide (poly(Py)) tracts and minimal Saccharomyces cerevisiae autonomously replicating sequence (ARS) consensus, in abundance comparable with that of these sequences of DNA fragments enriched in oris. All these DNAs were found to be enriched also in sequence elements held as primase (Pr) attachment sites. Moreover, DNAs of POMS component B and those of AMV DNA were found to be enriched in the asymmetric pyrimidine (Py) heptanucleotide motif of Waltz et al. (1996) occurring in the initiation zones of ori region. Consequently, these extrachromosomal DNAs, portion of which represents a precursor of AMV DNA, seem to descent from initiation zones of various ori regions of an early replicating chromosomal myeloblast DNA. In addition, a possible explanation of the inclination of these DNAs to form multimers is presented. PMID- 10672339 TI - Resistance to Friend leukemia virus in transgenic mice expressing the native Fv-4 gene. AB - Fv-4 is a truncated ecotropic retrovirus gene that codes for an envelope protein under control of a cellular promoter. It confers resistance to ecotropic murine leukemia viruses. Transgenic mice were derived using the native Fv-4 gene as the construct for microinjection. Two founder mice were derived. In both founder lines, there was no detectable expression of the transgene or resistance to Friend murine leukemia virus (FrMLV) in hemizygotes. In one line, the resistance was observed in homozygotes with Fv-4 RNA formation in the thymus but not in the spleen or in other tissues. In the other founder line, a homozygous male was identified. Double integrants, derived from breeding this homozygous male to homozygous females from the other founder line, were also resistant. These results indicate that the native gene confers the resistance in homozygous transgenic mice or double integrants derived from different founders but not hemizygotes. PMID- 10672340 TI - A Fe(3+)/DNA complex induces an anti-human immunodeficiency virus factor(s) in CD4+ lymphocyte cell lines. AB - Numerous cytokines and chemokines are involved in inflammatory and immune response. Whereas some of them inhibit virus replication in vitro directly or increase the patients' T4-lymphocyte level, others effects are not so clear. Using human immunodeficiency virus (HIV) and cell cultures we have studied the antiviral effect of complexes of salmon DNA with metals and of a new factor(s) (antiviral factor, AVF) induced in cells by the complexes. The Fe3+/DNA complex possessed the highest antiviral activity. It was found that MT-2, MT-4, CEM and Jurkat cells treated with the complexes secreted AVF which inhibited the replication of nine HIV-1 isolates, was noncytotoxic and stimulated cell proliferation. AVF did not inactivate HIV. The molecular mass analysis of AVF showed that its antiviral activity is associated with its fraction of M(r) of 3 K. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of mRNA from MT-4 cells treated with the complexes showed an increase in the the expression of genes for interleukin-1 alpha (IL-1 alpha), tumour necrosis factor alpha (TNF alpha) and TNF-beta while expression of genes for IL-1-beta, IL-2, IL-4, IL-6, IL 8. IL-10, IL-12; 35p, 40p, IL-13, GMCSF, GSF and RANTES was not detected at all. However, the anti-HIV activity of the cell culture supernatant in vitro cannot be explained by mere presence of the inflammatory substances mentioned above, because they do not possess such activity and their M(r) is higher than that of AVF. Our findings raise the possibility that AVF(s) may be involved in the mechanism of cell resistance against HIV. PMID- 10672341 TI - Accumulation of helper component/proteinase and coat protein of turnip mosaic virus in intact plants. AB - The helper component/proteinase (HC/Pro) protein of turnip mosaic virus (TuMV) was fused with glutathione S-transferase (GST) and expressed as a fusion protein in Escherichia coli. The quality of antiserum raised against the GST-HC/Pro fusion protein was compared to that of antiserum raised against coat protein (CP) by image analyser. The result showed that these antisera were of similar quality. Then the both antisera were used to follow the time course of accumulation of HC/Pro protein and CP in intact TuMV-infected leaves. CP appeared first at day 3 post inoculation (p.i.) and gradually accumulated in uninoculated upper leaves, whereas HC/Pro protein appeared first at day 4 p.i., accumulated up to day 7 p.i. and then gradually decreased. Potyvirus proteins are encoded by a single translation unit spanning most of the genome and are presumably synthesized in equimolar ratios. Therefore, the reduced accumulation of HC/Pro protein in relation to CP at one month p.i. in infected plants is presumed to be the result of its degradation. PMID- 10672342 TI - Interaction of V-Myb oncoprotein with spread chromatin of avian haematopoietic cells. AB - Interactions of v-Myb oncoprotein with spread chromatin of avian LSCC-BM2 cells expressing v-myb oncogene were studied by means of immunoelectron microscopy. The application of this technique using anti-Myb polyclonal antibody combined with the Miller type spreading for visualisation of chromatin revealed the presence of Myb protein on stretched chromatin fibres. Intense labelling was apparent on the chromatin dispersed by hypotonic treatment, where the label was present frequently in clusters, although individual marks along the fibrillar molecules were also found. The combination of hypotonic and detergent treatment resulted in better dispersal of chromatin, more frequent detection of active transcription units, but also in removal of some proteins from chromatin fibres. The labelling of chromatin with anti-Myb antibody was substantially reduced in this case and was dependent on detergent concentration used. The marker was found less frequently on chromatin fibres usually present in clusters on remaining protein structures. Our findings confirmed direct interaction of v-Myb protein with chromatin structure. This interaction is apparently affected by detergent treatment. PMID- 10672343 TI - Detection of single-stranded cohesive ends in the genome of Bacillus thuringiensis temperate phage KK-88. AB - Cohesive ends (cos sites) were detected in the genome of temperate KK-88 phage in Bacillus thuringiensis after analysis of the phage DNA generated by the 3'-5' exonuclease activity of Klenow fragment of DNA polymerase I. In addition, unlike the 5'-protruding ends of coliphage lambda genome, the ends of KK-88 phage genome were found to be 3'-protruding. The restriction map of the phage genome was also constructed on the basis of position of the cos fragments. PMID- 10672344 TI - A note on outbreaks caused by mixed foot-and-mouth disease virus infections. AB - Two outbreaks of foot-and-mouth disease (FMD) in vaccinated cattle were investigated wherein a mixed infection due to FMD virus (FMDV) types O and Asia 1 was detected by sandwich enzyme-linked immunosorbent assay (ELISA) and confirmed by antigen capture polymerase chain reaction (PCR). The clinical picture and the epidemiological data on these outbreaks are presented. The isolated virus strains were compared to the respective vaccine strains by means of monoclonal antibody (MAb) profiling and nucleotide sequence analysis. The probable cause of the mixed FMDV infection and its significance in disease control are discussed. PMID- 10672345 TI - Genetic homogeneity of human serotype G1 rotaviruses isolated during a single epidemic season: implications for vaccine strategies. AB - The extent of genetic variation of rotavirus isolates representing the common serotype G1 circulating in Australian urban centres during 1996 was investigated. The sequences of the major outer capsid glycoprotein, VP7, of three isolates from Brisbane, Sydney and Melbourne were determined and found to be highly conserved (> 99% nucleotide identity, > 98.3% amino acid identity and > 99% amino acid similarity). In particular, the sequences of the neutralisation epitope regions were absolutely conserved. These strains and those from other cities were analysed by Northern blot hybridisation using a probe derived from a prototype serotype G1 virus. For all strains, the eleven genomic RNA segments hybridised with the probe indicating that these strains were not derived from genetic reassortment between different rotavirus geno-groups. These results suggest that, within a single epidemic season, rotavirus strains circulating in geographically diverse communities share common genetic features. PMID- 10672346 TI - Nucleotide sequences of the cylindrical inclusion protein genes of two Japanese zucchini yellow mosaic virus isolates. AB - The nucleotide sequences of the cylindrical inclusion protein (CIP) genes of two Japanese zucchini yellow mosaic virus (ZYMV) isolates (ZYMV-169 and ZYMV-M) were determined. The CIP genes of both isolates comprised 1902 nucleotides and encoded 634 amino acids containing consensus nucleotide binding motif. The sequence similarities between the two isolates at the nucleotide and amino acid levels were 91% and 98%, respectively. When the CIP gene sequences of the Japanese ZYMV isolates were compared with those of previously reported ZYMV isolates, the nucleotide and amino acid sequence similarities ranged between 81% and 97%, and between 95% and 97%, respectively. Phylogenetic analysis of the deduced amino acid sequences of the CIP genes indicated that the Japanese ZYMV isolates were closely related to those of other ZYMV isolates. PMID- 10672347 TI - Immunogenicity of interferon-alpha 2 in therapy: structural and physiological aspects. AB - Recombinant human interferon (rIFN)-alpha 2 has been approved for therapeutic application in a range of human oncological and viral diseases. However, some patients can develop strictly specific antibody response to rIFN-alpha 2, which may diminish its therapeutic potential. Such humoral response appears to be quite complex and obviously depends on multiple parameters. Our review is aimed primarily to factors associated with structural modifications of rIFN-alpha 2 that we consider crucial for formation of therapy-induced antibodies. These factors are either related to inherent conformational differences between three IFN-alpha 2 subvariants or to immunogenically active contaminating derivatives resulting from production, purification and storage of this recombinant protein. In addition, the role of treatment regimen and physiological variables modulating the immune response to rIFN-alpha 2 in the challenged organism are mentioned. PMID- 10672348 TI - [Stoke: acute emergency, not fatal disease]. PMID- 10672349 TI - [Preclinical treatment of patients with stroke]. AB - Stroke is the third leading cause of death and number one cause of disability in industrialised countries. Studies into the pathophysiology of acute ischaemic stroke have indicated that treatment options are likely to be optimized when early signs of stroke are recognized and treatment is initiated within 3 hours from symptom onset. Therefore, new conceptions heading towards early diagnosis, fast preclinical treatment, structured diagnostics, immediate initiation of acute therapy as well as early initiation of rehabilitation are required. It is well known that, for most patients, there is a long delay between the onset of symptoms and the start of therapy. Many factors are responsible for the time delay:signs and symptoms often go unrecognized and/or are minimized by patients, relatives and bystanders. Unlike trauma or myocardial infarction, stroke is not given a high priority by medical staff and/or emergency medical services (EMS). Although a small number of stroke patients is treated as emergency and attended to by the emergency medical services within this time window, this number could easily be increased by intensified public and emergency personnel education. At present the standard of care by the EMS personnel includes adequate cerebral oxygenation, treatment of cardiac arrhythmia, management of hypertension as well as therapy of hyperglycemia and hyperthermia. For the future, we hope that emergency medical services will be able to initiate therapies which must be administered within the first few hours of acute stroke after onset of symptoms. Early notification of hospitals would enable a particular stroke team to be present at the patient's admission. PMID- 10672350 TI - [Respiratory pattern and respiratory strain in automatic tube compensation and inspiratory pressure support]. AB - STUDY OBJECTIVE: To investigate whether automatic tube compensation (ATC) or conventional pressure support (PS) is suitable to compensate for the work of breathing imposed by the breathing circuit without altering the breathing pattern. METHODS: Breathing pattern and work of breathing were measured in healthy volunteers. After a 20 min period of quiet breathing through a mouth piece (control) the volunteers were breathing through a 8.0 mm ID endotracheal tube (ETT) with four different settings: CPAP at 0 mbar, ATC, PS 5 mbar, PS 10 mbar. Each mode was applied for a 20 min period. At the end of each period data from 10 consecutive breaths were analyzed and averaged. Tidal volume (VT), breathing frequency (f), and minute ventilation (Ve) were determined from the stored gas flow tracings. Work of breathing was assessed as the pressure time product (PTP) calculated from the transdiaphragmatic pressure (Pdi) using a combined esophageal and gastric balloon catheter. RESULTS: During the control period the breathing pattern was as follows: VT = 882 +/- 277 ml, f = 13.7 +/- 5/min, Ve = 11.5 +/- 4.2 L/min. Maximal Pdi was 9.2 +/- 5.4 mbar and PTP was 11.3 +/- 7.1 mbar x s. Breathing CPAP through the ETT resulted in a slight increase in Pdi (10.8 +/- 5.4 mbar) and PTP (14.8 +/- 10.4 mbar x s) with an unchanged breathing pattern. However, for the same amount of unloading from respiratory workload ATC did not alter the breathing pattern, whereas PS 5 mbar and PS 10 mbar resulted in a clear increase in VT (1014 +/- 202 ml, 1336 +/- 305 ml, respectively). CONCLUSION: From the presented data in healthy volunteers it might be concluded that ATC and PS 5 mbar and 10 mbar are suitable modes for unloading the respiratory system from work imposed by the breathing circuit. ATC does not alter the breathing pattern in contrast to PS which results in an increased tidal volume. Therefore, the exact compensation of the work imposed by the ETT during ATC seems to be advantageous over ATC to assess the actual breathing pattern. PMID- 10672351 TI - [Intraoperative blood requirements and allogeneic blood transfusion in cardioanesthesia. Data analysis of 7729 patients in 12 cardiac surgical clinics]. AB - Allogeneic blood requirements in cardiac surgery shows a wide variation even for comparable procedures. The aim of the present study was to compare the intraoperative allogeneic blood requirement in defined cardiac operations among 12 cardiac centers in Germany. METHOD: A data set with 25 variables concerning the intraoperative course in adult cardiac patients with myocardial revascularization, valve replacement (aortic or/and mitral valve) or combined procedures was distributed to the participating centers. The data of all patients between January 1th 1998 and June 30th 1998 were included. Besides demographic data, the intraoperative transfusion of allogeneic and autologous blood, fresh frozen plasma and the concomitant hematocrit values were registered. Data were analyzed for all centers and separated for each center. RESULTS: The data of 7,729 patients were analyzed. The intraoperative allogeneic blood requirement was 0.6 +/- 1.3 units for all patients. It varied among the centers from 0.25 +/- 0.6 units to 0.97 +/- 1.6 units (P < 0.05). The percentage of patients receiving allogeneic blood was 27% and differed among the centers from 17% to 35%. Female patients were transfused in 53% (36-39%) compared to male patients with 16% (9 20%) (P < 0.05). The rate of autologous blood predonation varied from 0.5% to 23%. Patients without autologous predonation were transfused in 28% compared to 4% in patients with predonation (P < 0.05). In patients with autologous predonation the intraoperative transfusion of allogeneic blood was significantly reduced (0.1 +/- 0.39 vs 0.6 +/- 1.4 units, P < 0.05). However, some centers with a high percentage of autologous predonation also demonstrated a high rate of perioperative allogeneic transfusion. CONCLUSION: The incidence of allogeneic blood transfusion in cardiac surgery depends on the institution and not on the surgical procedure. A common threshold value of hemoglobin for the transfusion of blood trigger even for comparable procedures could not be detected among the centers. Especially in female patients, there was a wide variation in allogeneic blood transfusion. Autologous blood predonation reduces blood requirement significantly, however, it is practiced with variing intensity. The data set did not include information about transfusion regimen in the postoperative period, thus, these data do not allow to draw conclusions for the whole perioperative period. PMID- 10672352 TI - [Postoperative cognition disorders in elderly patients. The results of the "International Study of Postoperative Cognitive Dysfunction" ISPOCD 1)]. AB - OBJECTIVE: Cognitive dysfunction is a known problem after operations and may be especially relevant in the elderly. The aim of this international multicentre study was to investigate short- and long-term cognitive dysfunction in elderly patients and to elucidate the relevance of hypoxaemia and hypotension as causative factors. METHODS: 1218 patients aged 60 years and older and scheduled for major non-cardiac surgery under general anaesthesia were investigated. Oxygen saturation was measured by continuous pulse oximetry before surgery and throughout the day of and the first 3 nights after surgery. Blood pressure was recorded every 3 minutes during the operation and every 15-30 min for the rest of that day and night. Cognitive testing was performed before and 1 week and 3 months after the operation. Cognitive dysfunction was identified with neuropsychological tests compared with controls recruited from the UK (n = 176) and the same countries as study centres (n = 145). RESULTS: Postoperative cognitive dysfunction was present in 25.8% of patients 1 week after surgery and in 9.9% 3 months after surgery, compared with 3.4% and 2.8%, respectively, of the UK controls. Increasing age and duration of anaesthesia, little education, a second operation, postoperative infections, and respiratory complications were the risk factors for early postoperative cognitive dysfunction, but only age was a risk factor for long-term postoperative cognitive dysfunction. Hypoxaemia and hypotension were not significant risk factors at any time. CONCLUSION: With this investigation long-term cognitive dysfunction could be proven definitively for elderly patients after major operations under general anaesthesia. No factors with prophylactic or therapeutic influence were detectable so that aetiology and pathophysiology of POCD could not be further determined. PMID- 10672353 TI - [Inadvertent potassium chloride infusion in an epidural catheter]. AB - In a 65 years old male patient 38 cc of a 7.45% potassium chloride-solution was inadvertently infused within 3 hours into an epidural catheter on the first postoperative day. The epidural potassium chloride administration resulted in a paresis and painful paraesthesia of the patient's legs and a level of sensory blockade to TH 11. Furthermore vegetative symptoms like hypertension and tachycardia were observed. For therapy a single bolus of 40 mg dexamethasone was administered intravenously followed by an epidural infusion of sodium chloride 0.9% 99 cc/h for several hours. About 6 hours after the start of infusion all symptoms had disappeared. It is proposed that the use of colour-coded epidural catheter devices and coloured electrolyte solutions as well as infusion-pumps with a larger reservoir that reduce the frequency of syringe changes would be helpful in avoiding such complications. PMID- 10672354 TI - [Bilateral hydrothorax with hydromediastinum after accidental catheter dislocation]. AB - We report a complication during the insertion time of a central venous catheter in a patient with intracerebral bleeding. This complication was caused by an inadvertent dislocation of a subclavian catheter. Hydromediastinum and bilateral hydrothorax developed. There was a clear diagnosis followed by removal of the central venous catheter after radiological investigations could explain the cause of the complication and the clinical symptoms. In the course of events mediastinitis was diagnosed. The clinical condition improved under antibiotic therapy. The patient could be transferred to another clinical unit for endovascular treatment of an arterio-venous cerebral malformation. PMID- 10672355 TI - [Is the laryngeal mask a minimally invasive instrument for securing the airway? Supplementary remarks on the paper "Injures and dangers in the use of the laryngeal mask" by V. Hempel, Anaesthesist (1999)48:399-402]. AB - Minor laryngopharyngeal complaints following anaesthesia considerably determine postoperative patient comfort. They cannot be eliminated but reduced by experience and careful preparation and insertion technique. The incidence of minor laryngopharyngeal symptoms following the use of the laryngeal mask airway is similar to that following endotracheal intubation. However, there seems to be a distinct pattern of complaints: discomfort with swallowing is more frequent after LMA, whereas dysphonia is more often observed following endotracheal intubation. The significance of LMA cuff pressures in the pathogenesis of postoperative throat complaints remains unclear. There is sound evidence that cuff pressure is not a representative measure for the effective pressure load upon the pharyngeal mucosa. Measurement of cuff pressure is not obligatory, instead reduction of cuff volume to a "just seal" situation seems to be a reasonable approach. The laryngeal mask airway has definitely changed anaesthesiology airway management. Whether this is due to its supposedly less invasiveness compared to endotracheal intubation has not been proven by scientific investigations. PMID- 10672356 TI - [Cost control in the hospital. An introduction to cost and performance management]. PMID- 10672357 TI - [Excitation following sevoflurane: a problem in pediatric anesthesia?]. PMID- 10672358 TI - [The history of the development of intensive care in Germany--contemporary reflections. 12. The development of intravenous infusion techniques from personal experience]. PMID- 10672359 TI - [The implementation of the International Classification of Diseases (ICD-10) in Germany. Tools and information on the internet]. AB - ICD-10 is to be implemented for morbidity coding in Germany soon. The electronic versions of ICD-10 are introduced. For everyday work with the classification and for epidemiological research further tools are provided by DIMDI:ICD-10 meta files, ICD conversion tables, ICD-10 thesaurus of diagnostic terms. All files are available free of charge via the INTERNET. PMID- 10672360 TI - [Perioperative hypothermia]. PMID- 10672361 TI - A comparative Golgi study of chicken (Gallus domesticus) and homing pigeon (Columba livia) hippocampus. AB - Neurons and fibres in the chick and homing pigeon hippocampus were described following Golgi impregnation. Two principal classes of neurons were distinguished: projection neurons with distant projecting axons and spiny dendrites, and local circuit neurons. In the homing pigeon and chicken hippocampus there are three types of projection neurons: pyramidal, pyramidal like and multipolar. The pyramidal and pyramidal-like neurons are only found in the central 'pyramidal' layer of the hippocampus whereas multipolar neurons are present in the suprapyramidal, pyramidal and infrapyramidal layers. The axon of projection neurons typically emits several varicose collaterals from the initial section. Most of these collaterals extend along the infrapyramidal layer of the hippocampus, while others ascend to the pyramidal and suprapyramidal layers where they branch. The number of impregnated axon collaterals was higher in the homing pigeon than in the chick hippocampus. A variety of multi-angular/ovoid local circuit neurons ranging from small to large size are found in the homing pigeon and chick hippocampus. Their axons develop local arborization of varicose branches, the extent of which varies with the type of local circuit neurons. The density of GABA immunopositive local circuit neurons was found to be greater in the homing pigeon than in the chick. The profuse arborization of projection neuron axon collaterals and the higher density of GABA-immunopositive local circuit neurons in the homing pigeon hippocampus may underlie the differences in hippocampal function between the homing pigeon and chick, and this complex local connectivity may contribute to the ability of spatial orientation and memory. PMID- 10672362 TI - Immunolocalization of the transcription factor Slug in the developing avian heart. AB - Slug is a transcription factor involved in processes such as the formation of mesoderm and neural crest, two developmental events that imply a transition from an epithelial to a mesenchymal phenotype. During late cardiac morphogenesis, mesenchymal cells originate from two epithelia--epicardial mesothelium and cushion endocardium. We aimed to check if Slug is expressed in these systems of epithelial-mesenchymal transition. We have immuno-located the Slug protein in the heart of quail embryos between Hamburger and Hamilton stages HH16 and HH30. In the proepicardium (the epicardial primordium), Slug was detected in most cells, mesothelial as well as mesenchymal. Slug immunoreactivity was strong in the mesenchyme of the endocardial cushions and subepicardium from its inception until HH24, but the immunoreactivity disappeared in later embryos. Only a small portion of the endocardial cells located in the areas of epithelial-mesenchymal transition (atrioventricular groove and outflow tract) were immuno-labelled, mainly between HH16 and HH20. Endocardial cells from other cardiac segments were always negative, except for a transient, weak immunoreactivity that coincided with the development of the intertrabecular sinusoids of the ventricle. In contrast, virtually all cells of the epicardial mesothelium were immunoreactive until stage HH24. The mesenchymal cells that migrate to the heart through the spina vestibuli were also conspicuously immunoreactive. The myocardium was not labelled in the stages studied. Our results stress the involvement of Slug in the epithelial to mesenchymal transition. We suggest that Slug can constitute a reliable marker of the cardiac epithelial cells that are competent to transform into mesenchyme as well as a transient marker of the epithelial-derived mesenchymal cells in the developing heart. PMID- 10672364 TI - The formation of the chick ileal muscle layers as revealed by alpha-smooth muscle actin immunohistochemistry. AB - The genesis of intestinal smooth muscle layers was immunohistochemically investigated by use of an antibody to alpha-smooth muscle actin (alpha-SMA) in the developing chick ileum. Myoblast cells positive for alpha-SMA were already found in the presumptive circular muscle layer on E 8.5. On E 11.5 radially oriented muscle fibers were protruded from the outermost layer of the developing circular musculature and then formed a tuft-like aggregates. These radial muscle bundles were bent into an L-shape. The long distal extension of muscle bundles run parallel to the long axis of the ileal loop and developed into the longitudinal muscle layer. The obliquely oriented muscle fibers, locating at the intermuscular space of the muscularis propria, probably are to be considered a remnant of the short extension of radial muscle bundles. The muscularis mucosae was formed by the processes equivalent to the genesis of longitudinal muscle layer. On E 14.5 centripetally oriented muscle fibers emerged from the innermost layer of circular musculature. The long distal extension of centripetal fibers lay along the inner surface of developing circular musculature. On E 19.5 the longitudinal muscle layer of the muscularis mucosae was newly formed by separating from the circular musculature. The villous myoblast cells initially developed from the innermost layer of the muscularis mucosae on E 18.5, and were widely distributed in the lamina propria mucosae on E 20.5. Temporal and chronological pattern in expression of alpha-SMA was observed during the development of the chick intestinal smooth muscle. By E 14.5 the entire layer of the muscularis propria was intensely immunostained for alpha-SMA, but from E 15.5 onward the staining intensity gradually began to decrease from the outer half of the circular musculature. Finally, the immunoreactivity was localized in the inner layer of circular muscle and the longitudinal muscle layer. A possible functional role of this inner layer of circular muscle is discussed. PMID- 10672363 TI - "Rodent-like" and "primate-like" types of astroglial architecture in the adult cerebral cortex of mammals: a comparative study. AB - Previous observations disclosed that astroglia with interlaminar processes were present in the cerebral cortex of adult New and Old World monkeys, but not in the rat, and scarcely in the prosimian Microcebus murinus. The present report is a more systematic and comprehensive comparative analysis of the occurrence of such processes in the cerebral cortex of several mammalian species. Brain samples were obtained from adult individuals from the following orders: Carnivora (canine), Rodentia (rat and mouse), Marsupialia (Macropus eugenii), Artiodactyl (bovine and ovine), Scandentia (Tupaia glis), Chiroptera (Cynopteris horsfieldii and C. brachyotis), and Primate: Prosimian (Eulemur fulvus), non-human primate species (Cebus apella, Saimiri boliviensis, Callithrix, Macaca mulatta, Papio hamadryas, Macaca fascicularis, Cercopithecus campbelli and C. ascanius) and from a human autopsy. Tissues were processed for immunocytochemistry using several antibodies directed against glial fibrillary acidic protein (GFAP), with or without additional procedures aimed at the retrieval of antigens and enhancement of their immunocytochemical expression. The cerebral cortex of non-primate species had an almost exclusive layout of stellate astrocytes, with only the occasional presence of long GFAP-IR processes in the dog that barely crossed the extent of lamina I, which in this species had comparatively increased thickness. Species of Insectivora and Chiroptera showed presence of astrocytes with long processes limited to the ventral basal cortex. Interlaminar GFAP-IR processes were absent in Eulemur fulvus, at variance with their limited presence and large within- and inter-individual variability as reported previously in Microcebus murinus. In New World monkeys such processes were absent in Callithrix samples, at variance with Cebus apella and Saimiri boliviensis. Overall, the expression of GFAP-IR interlaminar processes followed a progressive pattern: bulk of non-primate species (lack of interlaminar processes)--Chiroptera and Insectivora (processes restricted to allocortex) < strepsirhini < haplorhini (platirrhini < catarrhini). This trend is suggestive of the emergence of new evolutionary traits in the organization of the cerebral cortex, namely, the emergence of GFAP-IR long, interlaminar processes in the primate brain. Interlaminar processes may participate in a spatially restricted astroglial role, as compared to the one provided by the astroglial syncytium. It is proposed that the widely accepted concept of an exclusively astroglial syncytium is probably linked with a specific laboratory animal species ("rodent-type" or, rather, "general mammalian-type" model) that misrepresents the astroglial architecture present in the cerebral cortex of most anthropoid adult primates ("primate-type" model), including man. PMID- 10672365 TI - Confocal microscopy of Tomes' granular layer in dog premolar teeth. AB - Tomes' granular layer is the hypomineralized area of radicular dentin, but knowledge concerning it is limited. The present study was designed to investigate the structural characteristics of Tomes' granular layer in the dog's teeth by confocal microscopy. Permanent premolars of four beagles, two at 7 months and the other two at 14 months of age, were used for observation. During premolar root formation, the 7-month-old dogs were injected with calcein and alizarin red S for vital staining of dentin, and ground sections of the teeth were prepared. Both ground and decalcified-paraffin sections were made from the teeth of the 14-month old dogs and stained with basic fuchsin or with hematoxylin and eosin. All sections were examined by fluorescence and confocal microscopy. In the ground sections, granules of Tomes' layer and dentinal tubules were stained with basic fuchsin and with calcein. The granules of Tomes' layer stained with calcein were seen only near the labeling lines by calcein. The granules of Tomes' layer appeared as bright spots in cross sections, and as lines in longitudinal sections. When the sections were cut tangentially through the surface of dentin, the granules of Tomes' layer showed a reticular structure. Most of the dentinal tubules were seen to pass between the granules and terminated in the dentin cementum junction. Looped tubules were not found in this area. In the paraffin sections stained with hematoxylin and eosin, extracellular matrix of dentin showed fluorescence of various intensities and dentinal tubules appeared dark. At the surface of the radicular dentin, the granules of Tomes' layer appeared as fluorescent fibers running parallel to the surface of dentin in the longitudinal sections. The fibers appeared as bright spots in the cross sections and as a mesh in the tangential sections. In the periodontal ligament, collagen fibers showed intense fluorescence, whereas most cells were negative. From these results we conclude that Tomes' granular layer of dog's teeth may be the collagen fiber bundles that remained uncalcified or hypocalcified within the radicular dentin. PMID- 10672366 TI - Development of full-length Trk B-immunoreactive structures in the prefrontal and visual cortices of the macaque monkey. AB - Distribution and morphological changes of cells containing the signal transducing neurotrophin receptor, full-length Trk B (fl-Trk B), were investigated in the prefrontal cortex (area FD) and the primary visual cortex (area OC) of the macaque monkey between embryonic day 140 and the adult stage. In area FD at the adult stage, fl-Trk B immunoreactivity was mainly observed in the pyramidal cells in layers II/III, V and VI. Small numbers of granule cells in layer IV were immunopositive. Bipolar and multipolar cells in layer II were rarely immunoreactive. At embryonic day 140, the number of fl-Trk B immunoreactive pyramidal cell was high, and gradually decreased until the adult stage. In layer IV, the number of fl-Trk B-ir cells was also high at embryonic day 140, and decreased remarkably from postnatal day 7 to the adult stage. On the other hand, in area OC at the adult stage, cells in layers II/III, IV, V and VI were fl-Trk B immunopositive. From embryonic day 140 until adulthood, the cells in layer IVc were fl-Trk B immunoreactive. The strongest fl-Trk B immunoreactivity in areas FD and OC occurred at postnatal month 6, coinciding with the time of the synapse overproduction. These findings suggest that ligands of fl-Trk B, such as BDNF and NT4/5 may be involved in the development and maintenance of the monkey cerebral cortices. PMID- 10672367 TI - Screening for surreptitious laxative abuse. PMID- 10672368 TI - Clinical governance. PMID- 10672369 TI - Principles, practice and paracetamol: when units matter. PMID- 10672370 TI - Pitfalls in forensic toxicology. PMID- 10672371 TI - Standardization of HbA1c measurements: a consensus statement. PMID- 10672372 TI - The clinical need for HbA1c standardization. PMID- 10672373 TI - Adult reference ranges for serum cystatin C, creatinine and predicted creatinine clearance. AB - Serum cystatin C measurement has been previously shown by ourselves and others to be a better indicator of changes in glomerular filtration rate (GFR) than serum creatinine. However, the available literature on reference values for cystatin C concentration remains surprisingly sparse; we thus set out to determine an adult reference range. Blood was taken from 309 healthy blood donors and creatinine and cystatin C concentrations were measured using commercially available automated methodologies. In addition, predicted creatinine clearances were calculated using the Cockcroft and Gault formula. The 95% reference intervals for creatinine, predicted creatinine clearance and cystatin C for all blood donors, regardless of gender, were 68-118 mumol/L, 58-120 ml/min/1.73 m2 and 0.51-0.98 mg/L, respectively. For women, the intervals were 68-98 mumol/L, 60-119 ml/min/1.73 m2 and 0.49-0.94 mg/L; for men, they were 78-123 mumol/L, 57-122 ml/min/1.73 m2 and 0.56-0.98 mg/L. This mean 95% reference interval for cystatin C in all donors under 50 years of age was 0.53-0.92 mg/L; for those over 50 years of age it was 0.58-1.02 mg/L. The small difference between make and female ranges meant that a single reference range for cystatin C could be established for all adults under 50 years of age without adjustment for body surface area. Serum cystatin C measurement offers a simpler and more sensitive screening test than serum creatinine for early changes in GFR. PMID- 10672374 TI - Glomerular filtration rate by single-injection inulin clearance: definition of a workable protocol for children. AB - The total serum clearance of inulin after a single intravenous injection is an established method for the determination of the glomerular filtration rate (GFR). To optimize this procedure in children, we analyzed 117 serum inulin time-decay curves in 59 children (age range 1.3-15.2 years)with renal disease. It was found that the two-compartment model fitted statistically better to the disappearance curves of serum inulin than the one-compartment model. The sampling frequency could be reduced from 12 to a minimum of seven data points (t=0, 10, 20, 30, 65, 120 and 240 min) in 4 h without a relevant change in the fit of the two compartment model to the date or in the calculated clearance of serum inulin. We conclude that, by using a standardized 'seven-sample schedule', followed by a two compartment analysis, the single-injection serum clearance of inulin is a suitable compromise between practical convenience and clinical accuracy for the determination of the GFR in children. PMID- 10672375 TI - Enzyme inhibitory antibody to pyruvate dehydrogenase: diagnostic utility in primary biliary cirrhosis. AB - In primary biliary cirrhosis, autoantibodies are produced to the family of 2 oxoacid dehydrogenase complexes. These 'anti-mitochondrial' antibodies are traditionally detected by immunofluorescence but this method of detection is subjective and labour-intensive. We assessed an enzymatic mitochondrial antibody (EMA) assay based on antibody inhibition of enzymatic activity of pyruvate dehydrogenase complex in wells of microtitre plates with a colorimetric read-out. We tested 48 Australian and 1947 Japanese patients with primary biliary cirrhosis, 306 normal subjects and 691 patients with various hepatic and non hepatic diseases. The overall sensitivity of the EMA for the diagnosis of primary biliary cirrhosis, 82%, was slightly lower than that of immunofluorescence, 90% The advantages of the EMA test include high specificity, >99%, and semi-automated features facilitating objectivity, rapidity, simplicity and economy. The EMA test could be particularly applicable to population screening for early primary biliary cirrhosis. PMID- 10672376 TI - Quantification of urinary oxalate with a chloride- and nitrate-insensitive oxalate oxidase. PMID- 10672377 TI - Hypertonic versus isotonic salt bridges and the measurement of ionized calcium. PMID- 10672378 TI - Calculation of renal tubular reabsorption of phosphate: the algorithm performs better than the nomogram. PMID- 10672379 TI - Enzymatic assay for magnesium in serum. PMID- 10672380 TI - Diagnostic dilemmas in Cushing's syndrome. PMID- 10672381 TI - Traumatic chylous ascites. PMID- 10672382 TI - A case of factitious hypercalcaemia. PMID- 10672383 TI - Glycation rates in type 1 diabetes mellitus. PMID- 10672384 TI - Creatinine clearance and glomerular filtration rate. PMID- 10672385 TI - A pilot study of the efficacy and tolerability of intralesional recombinant human beta-interferons in cervical intraepithelial neoplasia. AB - Beta-interferons possess anti-viral, cell proliferation inhibition and immunomodulatory characteristics which may be of use in the treatment of cervical intraepithelial neoplasia (CIN). Intralesional administration may avoid systemic side effects. Twenty-six women with cervical punch biopsy proven CIN I and II were treated by interferon injection into the cervical transformation zone according to three dosage regimens--6 million international units (IU) weekly for six weeks, 9 million IU weekly for six weeks and 12 million IU bi-weekly for three weeks. At the last treatment visit, cervical cytology and biopsy were taken to ensure no disease progression and large loop excision of the transformation zone (LLETZ) was carried out six months after treatment. Therapy was well tolerated with 93% of the scheduled 156 treatments given. Side effects which caused cessation of treatment included severe myalgia, headaches and prolonged flu-like symptoms. The 2 patients who failed to attend for LLETZ at six months and another 2 patients who received fewer than five scheduled treatments were excluded from analysis. LLETZ histology was negative in 12 patients (54%), showed inflammatory changes or squamous metaplasia in 4 (18%), was unchanged in 4 patients and had progressed in 2 (10%). Whilst intralesional beta-interferon clearly has activity in CIN, the response rate is less than that seen for excisional or ablative procedures. Nevertheless, it may have a role in the management of CIN where, for medical reasons of patient preference, there is a desire to avoid surgical therapy. PMID- 10672386 TI - The effect of fenofibrate on insulin sensitivity and plasma lipid profile in non diabetic males with low high density lipoprotein/dyslipidaemic syndrome. AB - Non-diabetic males, particularly those with very low levels of high density lipoprotein (HDL) type cholesterol and high levels of very low density type lipoprotein (VLDL), are associated with insulin resistance and decreased insulin sensitivity. The evidence that elevation of HDL cholesterol and diminution of triglycerides with drugs, with improvement in insulin sensitivity is still lacking. In the treatment of the dyslipidaemic syndromes with hypolipidaemic drugs, the associated metabolic abnormality of insulin resistance/sensitivity has to be addressed. We investigated the degree of decreased insulin sensitivity in 23 patients with low HDL and/or raised triglycerides by measuring the fasting, first and second hour insulin levels during an oral glucose tolerance test (OGTT) and repeated the measurements after a 6-month course of fenofibrate. The insulin levels were correlated with the OGTT, blood pressure, total cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides measured before and at the end of the trial. The serum insulin at the second hour of the OGTT fell from 100.79 +/- 42.79 mU/l to 54.56 +/- 25.43 mU/l (P < 0.0005) even though there was no change in the blood glucose level at this point. Our study shows that fenofibrate (Lipanthyl) 300 mg daily significantly raises the pretreatment low HDL cholesterol (from 0.64 +/- 0.1 mmol/l to 0.99 +/- 0.2 mmol/l) as well as lowers the triglyceride level (from 2.17 +/- 1.1 mmol/l to 1.43 +/- 0.64 mmol/l) in patients with low HDL/dyslipidaemic syndrome. The data also support the conclusion that treatment with fenofibrate increases insulin sensitivity as measured by the corresponding insulin levels of the OGTT in the study subjects who presented with very low HDL cholesterol level. There was also a decrease in blood pressure readings in our study subjects. Throughout the trial, there was no significant change in body weight or exercise level in the subjects studied. PMID- 10672388 TI - Excimer laser phototherapeutic keratectomy for recurrent corneal erosions. AB - The purpose of this paper was to review the Singapore National Eye Centre's (SNEC) experience with excimer laser phototherapeutic keratectomy (PTK) for treating recurrent corneal erosions (RCE). Thirty-five eyes of 32 patients who had PTK between 1992 and 1997 in SNEC were studied retrospectively. There was a history of previous ocular trauma in 15 eyes. Sixteen eyes had anterior corneal dystrophy. All had received conventional therapy without improvement in symptoms. The mean duration of symptoms prior to PTK was 19 months (range 1 to 71 months). PTK was performed with one of two 193 nm excimer lasers (Summit UV200LA, Summit Technology, Waltham, USA or Visx 20/20B, Visx Corp, Santa Clara, USA). An average of 30 laser pulses were delivered to Bowman's membrane after debridement of the corneal epithelium. The mean follow-up time was 12 months (range 0 to 56 months). Among those with adequate length of follow-up, 26/27 eyes (96%) were symptom-free for 3 months, 19/22 eyes (86%) were symptom-free for 6 months and 9/13 eyes (69%) were symptom-free for 1 year. Three eyes had repeat PTK. Mild corneal haze was seen in 3 eyes. No visually threatening complications were encountered. PTK is a safe and effective procedure for RCE refractory to conventional treatment. PMID- 10672387 TI - Use of EMLA cream or alfentanil for analgesia during ophthalmic nerve blocks. AB - This prospective double-blind study compared the effectiveness of EMLA with alfentanil and placebo in reducing the overall pain during ophthalmic nerve blocks. Seventy-five patients scheduled for cataract surgery were divided into three groups. Patients in the EMLA group had EMLA cream applied over skin areas corresponding to injection sites for retrobulbar and facial nerve blocks one hour before the nerve blocks, and placebo intravenous normal-saline injection 2 minutes before the first nerve block. The alfentanil group had placebo cream applied and intravenous alfentanil 10 micrograms.kg-1 while patients in the placebo group received placebo cream and intravenous normal-saline at similar time intervals prior to the nerve blocks. Patients then received facial nerve blocks and retrobulbar block by the same surgeon. Pain scores by patients and independent observers were significantly lower in the EMLA and alfentanil groups compared to placebo (P < 0.005) with no significant difference between the EMLA and alfentanil groups. PMID- 10672389 TI - Transscaphoid perilunate fracture/dislocations--results of surgical treatment. AB - Sixteen cases of dorsal transscaphoid perilunate fracture/dislocations in 15 patients were treated by open reduction and internal fixation of the scaphoid with a Herbert screw and/or Kirschner wires. All patients were male, with a mean age of 31 years. The average follow-up period was 3 years. These perilunate dislocations were transscaphoid in 13 cases, and transstyloid and transscaphoid in 3 cases. There was one case of median nerve deficit preoperatively. Open reduction was performed through a volar approach in all cases. Herbert screw fixation of the scaphoid was performed on 13 cases, of which supplemental Kirschner wires were used in 2 cases. Three cases had fixation with Kirschner wires only. There were 2 cases of non-union which required bone grafting on follow-up. A clinical evaluation scoring system assessing pain, occupational ability, range of motion and grip strength was used. Based on this, there were 10 excellent to good (62.5%) and 6 fair results. For the majority of our patients, surgical outcome is characterised by acceptable relief of pain, functional motion and grip strength. PMID- 10672390 TI - Perinatal drug abuse in KK Women's and Children's Hospital. AB - No local figures are available in Singapore on the incidence of perinatal drug abuse and its effect on the foetus and the neonate. The objectives of this study were to determine the incidence of perinatal drug abuse and neonatal abstinence syndrome; to identify a maternal profile at high risk for substance abuse and to document the presenting features and treatment of infants with neonatal abstinence syndrome. Out of 14,690 births during the period January 1994 to December 1996, 38 (0.25%) had evidence of perinatal drug abuse. The study revealed that a high-risk maternal profile for drug abuse comprised of single mothers (52%); history of smoking (52%); no antenatal care (37%) and belonging to the Malay ethnic group (82%); and younger maternal age. Self-reporting was uncommon, occurring only in 8% and in 40% of cases, there was no known history of maternal drug addiction. The drug abused in all cases was heroin. Human immunodeficiency virus (HIV) screening was done only in a minority (21%) of the mothers and it was negative in all. Eighteen (47%) infants had evidence of neonatal abstinence syndrome with neurological manifestations being the commonest. Urine toxicology screening was positive in 26% of cases and had only 70% sensitivity and 41% positive predictive value. On follow up, default rate was high with 42% babies not attending follow up at the outpatient clinic. In conclusion, there is a need to maintain a high index of suspicion of substance abuse in those with high-risk maternal profile and their neonates should be closely watched for features of neonatal abstinence syndrome. Alternative methods of toxicology screening apart from urine need to be evaluated in order to improve the drug detection rate. PMID- 10672391 TI - Is staging of benign prostatic hyperplasia (BPH) feasible? AB - With better understanding of the natural history of benign prostatic hyperplasia (BPH), the treatment can be tailored to the severity of the disease. The aims of this study were to determine the feasibility of staging BPH according to its severity and choose the optimal therapeutic tool for each category, and for comparing results of various modalities of treatment. Two hundred and twenty-five patients with clinical BPH were seen between October 1994 and July 1995. Initial assessment included the International Prostatic Symptom Score, and the quality of life index, digital rectal examination, urinalysis, prostate specific antigen, uroflow and residual urine estimation. Patients were then divided into: Stage 1, those with no bothersome symptoms and no significant obstruction, they can generally be watched. Stage 2, those with bothersome symptoms but without significant obstruction, they can be treated with pharmacotherapy/thermotherapy. Stage 3, those with significant obstruction defined as uroflow of less than 10 ml/s with persistent residual urine of > 100 ml, transurethral prostatic resection (TURP) would be recommended. Stage 4, those with complications of BPH such as chronic retention of urine and bladder stone, they would need TURP. One hundred and fifty-nine patients had complete follow up data of at least 2 years. Of the 70 patients who were originally in Stage 1, 59 (89%) remained in status quo, 6 patients developed acute retention of urine and only 1 required TURP. Of the 38 patients in Stage 2, 24 were down-staged to Stage 1 after medication and thermotherapy but 4 still remained in Stage 2 and the other 10 had worsening of symptoms requiring surgery. Of the 46 patients in Stage 3, 30 (65%) had TURP and all except 1 were down-staged to Stage 1. All patients in Stage 4 had TURP and improved. We conclude that staging of patients with clinical BPH is feasible. It serves as a useful guide for management and improves cost effectiveness. PMID- 10672392 TI - Focused abdominal sonography for trauma (FAST). AB - This study aims to examine the feasibility of applying the focused ultrasound examination (FAST) in the initial assessment of the trauma patient. Its advantage over diagnostic peritoneal lavage lies in the speed in which it can be performed, its non-invasiveness and repeatability. Over the period January 1997 to July 1998, the FAST examination (Acuson 128XP/10, 3.5 MHz probe) was used to assess the presence of haemopericardium or haemoperitoneum in 38 multi-trauma patients admitted to the Singapore General Hospital. The sample comprises mainly males (82%) with a mean age of 34 years (range 17 to 77 years). The mechanism of injury was predominantly blunt (95%). Mean Injury Severity Score (ISS) was 22. Eleven patients presented in shock and 16 patients had abdominal tenderness on examination. A single surgeon did the examination, which was performed during the secondary survey phase of resuscitation. Time taken for the FAST examination averaged 2.3 +/- 1.3 minutes. The results of the examination were compared to diagnostic peritoneal lavage, CT scan, operative findings, serial examination and/or post-mortem findings. Overall sensitivity was 67% and specificity 97%. Although the FAST examination missed the small amount of free fluid seen in the CT scan of 2 patients, these patients did not have to undergo laparotomy as their abdominal examination was normal. We conclude that the FAST examination is feasible and should be part of a general surgeon's armamentarium in the initial assessment of trauma. PMID- 10672393 TI - Outcome of tunnelled central venous haemodialysis catheters inserted by radiologists. AB - Radiologists have only recently been involved in the percutaneous placement of tunnelled central venous haemodialysis catheters. We report our initial experience with our first 60 catheters. All catheters were successfully inserted. Immediate complications encountered included puncture site haemorrhage in 3 patients (5%) and puncture of the left brachiocephalic vein in 1 patient (1.7%). These were managed conservatively without any clinical sequelae. About 80% of the catheters were uncomplicated and removed electively. Slightly more than 80% of the catheters were in place for more than 30 days. Infection and blocked catheters were the most common short-term complications. Ten catheters (17%) were infected resulting in premature removal of 9. There was 1 death from presumed line sepsis. Mean duration before the onset of infection was 53 days; the rate of infection was 0.28 episodes per 100 catheter days. Five catheters (8%) were blocked or had poor flow. The mean duration before the onset of blockage was 39 days and the rate of blockage was 0.14 episodes per 100 catheter days. A higher proportion of catheters inserted from the left encountered complications. In conclusion, percutaneous insertion of tunnelled haemodialysis catheters by radiologists is safe and effective. The right internal jugular vein should be the preferred access site. Precautions should be taken to avoid infectious complications given the high rate of catheter removal amongst infected catheters. PMID- 10672394 TI - Video-assisted thoracoscopy: role in the management of intrathoracic pathology. AB - A retrospective study was done from October 1992 to December 1996 of 48 patients who underwent video-assisted thoracoscopy. A male to female ratio of 3.7:1 was seen, with a mean age of 50 +/- 19 years. The underlying diagnoses included spontaneous pneumothorax (n = 23), malignant pleural effusion (n = 16), lung disease (n = 4), trauma (n = 2), empyema (n = 2) and oesophageal carcinoma (n = 1). The use of this modality is discussed in the treatment of various intrathoracic pathologies. PMID- 10672395 TI - An in vitro evaluation of epidural catheters: tensile strength and resistance to kinking. AB - The tensile strengths of 12 commercially available brands of epidural catheters were assessed using an Instron material testing device. The mean values of the tensile strengths ranged from 1.89 to 3.74 kilogram force. The extent of catheter occlusion due to kinking was also studied using an in vitro apparatus designed to simulate drug delivery at various degrees of flow restriction. It was determined that reinforced catheters were less likely to be occluded secondary to kinking. PMID- 10672396 TI - Repigmentation of vitiligo with autologous blister-induced epidermal grafts. AB - Treatment of vitiligo can sometimes be difficult and disappointing. PUVA treatments give fairly good results. However, acral regions like the hands or feet or areas over bony prominences like the elbow, are resistant to PUVA. Blister-induced epidermal grafts have been used to repigment vitiligo skin. This study was carried out on patients with vitiligo areas unresponsive to either PUVA treatments or who had segmental vitiligo. PUVA treatments were resumed after skin grafting. Twenty-five patients with stable vitiligo were grafted with blister induced epidermal grafts. Up to 70% of the whole vitiligo areas were grafted in one sitting. A total of 105 grafts were done. In 9 grafts, no repigmentation was seen. The remaining 96 grafts resulted in pigmentation. Twelve had partial and 84 had complete repigmentation. No Koebner phenomenon was noted in both the recipient sites or the donor sites. Blister-induced epidermal graft is an effective alternative to repigment stable vitiligo areas. It is easy to do and results are good. In this study, 96 out of 105 (91%) grafts had repigmentation. PMID- 10672397 TI - One-year review of pityriasis rosea at the National Skin Centre, Singapore. AB - Pityriasis rosea is a common, acute, self-limited papulosquamous eruption of possible viral aetiology. The aim of this study was to describe the profile of pityriasis rosea seen at a referral skin centre in Singapore. A retrospective chart review was conducted of all the patients with a diagnosis of pityriasis rosea seen during 1996. There were 368 patients; their ages ranged from 9 months to 82 years with a peak in the 20 to 29 years age group. There were slightly more males (1.2:1). The clinic incidence was 6.5 per 1000 cases. No ethnic predilection was noted and cases were seen evenly throughout the year. About a quarter of the patients gave a history of a viral infection shortly before or during the occurrence of the rash. Most cases had a typical truncal distribution. The herald patch was observed in 63 patients (17%). Inverse distribution involving mainly the extremities were seen in 22 cases (6%) and eczematised lesions were noted in 20 cases (5.4%). The two main differential diagnoses considered were tinea infection and secondary syphilis. A fungal scrape and a rapid plasma reagin test were done in 58 and 59 patients, respectively, and the results were negative. Treatment was symptomatic and consisted of topical steroids and antihistamines. Thirty patients (8%) were given oral prednisolone for extensive eruptions. The pattern of pityriasis rosea in Singapore is similar to that reported in other countries except for a male predominance and absence of monthly variation. A lower incidence and an older group of patients were also seen in our series as compared to African patients. PMID- 10672398 TI - Intracoronary brachytherapy: the beginning of the end of restenosis? AB - Restenosis remains a major problem 20 years after the introduction of coronary angioplasty. Pharmacological therapy has generally been disappointing in reducing the incidence of neointimal hyperplasia. Intracoronary brachytherapy is the latest anti proliferative agent that is showing promise in the fight against restenosis. Radiation therapy has been used for decades in the treatment of benign proliferative disorders. Animal studies have consistently demonstrated the efficacy of radiation therapy in inhibiting neointimal hyperplasia. Clinical trials, using different radioactive isotopes and radiation sources, are now showing dramatic reduction in neointimal hyperplasia and restenosis rates following balloon angioplasty and stenting. Intracoronary brachytherapy is beginning to show promise as one of the truly effective agent against neointimal hyperplasia and restenosis. PMID- 10672399 TI - Somatisation among Asian refugees and immigrants as a culturally-shaped illness behaviour. AB - Epidemiological studies indicate a high prevalence of major depression and anxiety disorder (including post-traumatic stress disorder) among Asian refugees and immigrants living in North America. Yet there exists an alarming underutilization of mental health services and underdiagnosis of psychiatric illness in this rapidly growing minority group. In order to investigate a culturally-derived basis for these observations, a critical review was conducted on descriptive epidemiologic, sociologic, and anthropologic studies of psychiatric illness among Asians and Asian refugees and immigrants reported in the general psychiatric and trans-cultural psychiatric literature of the past forty years. Studies examining the mode of illness presentation among Asian refugees seeking medical care suggest a marked tendency to articulate somatic rather than affective complaints when serious underlying psychiatric conditions exist. In this context, somatisation among Asian refugees and immigrants may reflect culturally-shaped beliefs regarding notions of disease aetiology and treatment as well as what is deemed culturally-appropriate help-seeking behaviour during illness. Misdiagnosis and underdiagnosis of psychiatric illness in this and other minority populations can be minimised by establishing pluralistic norms and multidimensional criteria which take into account the ethnically diverse manifestations of illness behaviour encountered increasingly in Western primary care and psychiatry clinics. PMID- 10672400 TI - Atypical mycobacterium infection with sporotrichoid spread in a patient with human immunodeficiency virus. AB - A case of Mycobacterium marinum infection presenting with a sporotrichoid spread in a HIV positive Chinese male is presented. The patient responded to oral cotrimoxazole treatment. A brief review of the literature concerning atypical mycobacterial infection presenting in such a fashion is discussed. PMID- 10672402 TI - Bullous dermatomyositis associated with nasopharyngeal carcinoma--a case report. AB - Cutaneous manifestations of dermatomyositis commonly include Gottron's papules, heliotrope rash, photosensitivity, poikiloderma and nailfold telangiectasia. Vesicles and bulla are rare. We report a patient with dermatomyositis who presented with blisters and oral ulcers. It is important to recognise this bullous variant in order to avoid a delay in diagnosis. Bullous dermatomyositis may also portend a poorer prognosis. Our patient was subsequently diagnosed to have undifferentiated nasopharyngeal carcinoma. PMID- 10672401 TI - Case reports of linear IgA bullous dermatosis of childhood. AB - Linear IgA bullous dermatosis of childhood (LADC) is an uncommon acquired blistering skin disorder which affects young children. It is characterised by a linear band of IgA at the epidermal basement membrane on direct immunofluorescence. From 1984 to 1988 at the previous skin centre at Middle Road Hospital, and 1989 to 1997 at the present National Skin Centre, a total of 4 cases were seen. All the patients were female and their ages ranged from 5 to 14 years (mean 8 years). There were 2 Chinese, 1 Caucasian and 1 Malay. All presented with an acute onset of blistering of 1 to 3 weeks' duration. Their skin biopsies showed subepidermal bullae with neutrophils, with/without eosinophils and direct immunofluorescence tests revealed linear deposits of IgA along the dermoepidermal junction. All were treated with prednisolone and dapsone with good results. One patient developed dapsone-induced haemolysis and treatment was changed to colchicine. We review the epidemiology, aetiology, current concepts and treatment modalities of this condition. PMID- 10672403 TI - Chryseobacterium meningosepticum (Flavobacterium meningosepticum)--a report of five cases in a local hospital. AB - Chrysobacterium meningosepticum (Flavobacterium meningosepticum) is a known cause of meningitis in premature and newborn infants. Infection due to this organism in adults is uncommon. We report 5 cases of Chrysobacterium meningosepticum in adult patients. Most of these patients were elderly and had underlying co-morbidities. PMID- 10672404 TI - Chronic subdural haematoma presenting with transient ischaemic attacks--a case report. AB - We report a middle-aged man who presented with repeated episodes of transient ischaemic attacks (TIA) in the setting of a chronic subdural haematoma. This case report discusses the various pathophysiologic mechanisms whereby such TIA may occur in chronic subdural haematoma. We also highlight the importance of cranial imaging in cases of TIA. PMID- 10672405 TI - Haemorrhage into non-functioning adrenal cysts--report of two cases and review of the literature. AB - Adrenal cysts are a rare condition and are usually non-functioning and asymptomatic. Most of the reported cases were incidental findings or discovered at autopsy. However, large cysts have a tendency to develop complications such as intracystic haemorrhage and rupture, which can present as an acute surgical emergency. We report two cases of adrenal cysts with intracystic haemorrhage. One patient presented with persistent non-specific upper abdominal pain, investigations with ultrasound (US) scan and computed tomographic (CT) scan revealed a left adrenal cyst and gallstones. Simultaneous cholecystectomy and adrenalectomy was performed with resultant relief of symptoms. The second patient presented with acute abdominal pain simulating acute surgical abdomen. Preoperative CT scan showed a large cystic lesion in the region of the tail of the pancreas with radiological evidence of haemorrhage but was unable to confirm its origin. The cyst was found to have arisen from the left adrenal gland at laparotomy; left adrenalectomy with complete excision of the cyst was done. Histology showed pseudocyst with haemorrhage in both cases. Pseudocyst is the commonest histological type encountered clinically. We believe the second case is related to pregnancy and childbirth as the patient presented during puerperium and the cyst, even though very large in size (25 x 15 x 15 cm), was not noted during antenatal screening with US scan. PMID- 10672406 TI - A case report of adult T-cell leukaemia/lymphoma (ATLL). AB - Adult T-cell leukaemia/lymphoma (ATLL) is a unique disease with distinct manifestations, a characteristic phenotype and a likely retroviral aetiology. It is unusual in Southeast Asia, though more commonly seen in some countries like Japan. We report a case of this disease in a 71-year-old man in Singapore who presented with papular erythematous eruptions for 6 years and subsequently developed generalised lymphadenopathy. He was diagnosed to have ATLL and, despite an initial response, became resistant to combination chemotherapy. We discuss the aetiology, characteristics and management of this interesting disease. PMID- 10672407 TI - Incessant ectopic atrial tachycardia and tachycardia-related cardiomyopathy: therapeutic options and potential for cure. AB - Incessant ectopic atrial tachycardia (IEAT) is a rare cause of cardiomyopathy. Cardiomyopathy is reversible by curative ablation using surgery or radiofrequency current. We report our experience with 5 patients with IEAT. Three patients presented with palpitations and were diagnosed to have paroxysmal supraventricular tachycardia (2 patients) and atrial flutter with 1:1 conduction (1 patient), but 2 presented insidiously with congestive cardiac failure. All the initial echocardiograms showed left ventricular dysfunction. The patients underwent electrophysiological studies which confirmed the diagnosis of IEAT. The first patient had surgical cryoablation and the other patients had successful radiofrequency catheter ablation. Follow-up for 2 to 7 years has shown no recurrences. All patients had significant improvement in left ventricular function on echocardiography. In conclusion, curative ablation by surgery or radiofrequency current is safe and effective. Because of its low morbidity, radiofrequency catheter ablation should be the treatment of choice for IEAT, especially if complicated by tachycardia-related cardiomyopathy. PMID- 10672408 TI - Reconstruction of a supinated hypoplastic thumb with combined Huber transfer and derotation osteotomy: a case report. AB - The use of the abductor digiti minimi transfer to restore opposition in patients with hypoplasia of the thumb has been widely described in the literature. It has been found to be effective in restoring abduction, but less so in restoring the rotational component of opposition. In cases where there is concomitant supination of the thumb, abductor digiti minimi transfer alone would not result in a good pinch as the thumb pulp is rotated away from the opposing finger. We present a case of a 6-year-old girl with a hypoplastic supinated left thumb which resembled a digit. There was also hypoplasia of the index finger. The hand had poor function as a result of lack of opposition of the thumb. The thumb function was restored by combining a derotational osteotomy (80 degrees) with the abductor digiti minimi transfer originally described by Huber. Patient was able to hold small object using key pinch and she could pinch with opposition of the thumb pulp to the middle, ring and little finger pulps when reviewed 2 years post surgery. PMID- 10672409 TI - Osteoid osteoma after a stress fracture of the tibia: a case report. AB - A 24-year-old man presented with a stress fracture over his left tibia following an infantry training 6 years ago. X-rays of his left tibia revealed a stress fracture and bone scan showed marked tracer uptake at the fracture. He was treated conservatively but his pain persisted since then. Five years later, X rays and histological examination confirmed the diagnosis of osteoid osteoma. The stress fracture may act as a trigger for the formation of osteoid osteoma and caused a delay in diagnosis. PMID- 10672410 TI - Multivariate statistical analysis: a brief introduction. PMID- 10672411 TI - 9th Seah Cheng Siang Memorial Lecture: gastric cancer--where are we now? AB - Gastric cancer, the second most common cancer in the world, kills about one million people a year, almost half of whom are Chinese. Chinese, Japanese and Koreans as well as east Europeans top the list with over 40 per 100,000 population per year, with a wide margin over Americans, Indians and Zimbabweans in whom the rates are below 1 per 100,000. The excellent prognosis of early gastric cancer is well established, and survival of cancer involving beyond the submucosa remains poor and there is little new in management. However, recent years have witnessed a breakthrough in the understanding of causative factors and molecular genetic abnormalities in gastric cancer that should pave the way for prevention, early detection and prognostication. Established carcinogens for gastric cancer now include Helicobacter pylori and N-nitroso compounds; other causative factors include salt and salted food intake, cigarette smoking, male sex, and familial genetic abnormalities. H. pylori infection increases cancer risk by about 5 in a 10-year period. Diet high in salt carries a relative risk of up to 6, and a highly significant correlation between 24 h urinary salt content and incidence of gastric cancer has been shown in 24 countries. The risk from smoking and male sex is under 2. Many N-nitroso compounds, which come from nitrites, which in turn come from nitrates in food following bacterial transformation in a hypochlorhydric environment, are established carcinogens in animals, but their risk for human gastric cancer is still debatable. The intestinal type of gastric cancer, according to Correa's hypothesis, develops from chronic inflammation leading to intestinal metaplasia, dysplasia and cancer, and is more associated with H. pylori and early gastric cancer. The diffuse type of gastric cancer does not go through these precancerous conditions and moves straight from inflammation to cancer. Associated with inflammation are an increase in proliferation and apoptosis, and this fine balance between proliferation and apoptosis may be uncoupled by genetic mutations. It is believed that as a result of the accumulation of molecular genetic abnormalities, a cancer eventually develops and metastasizes. p53 mutation, cyclin overexpression (especially in intestinal type), microsatellite instability, down regulation of E cadherin (especially in diffuse type), and telomerase reactivation are some prominent examples. These molecular abnormalities have the potential for screening, early detection and prognostication. Fruits and vegetables, green tea, alpha-tocopherol and other micronutrients such as selenium have been shown to reduce the risk for gastric cancer. In fact, it has been reported that diet consisting of vegetables and fruits, low in salt, together with the avoidance of cigarette smoking would prevent two-thirds to three-quarters of gastric cancer. Furthermore, eradication of H. pylori, and for that matter future vaccination, has the theoretical potential of preventing gastric cancer, and the potential use of COX2 inhibiting NSAID in inducing apoptosis may reverse precancerous conditions of the stomach. Both approaches are being intensely studied. PMID- 10672412 TI - Early 21st century professional practice: change and challenge. PMID- 10672413 TI - Current continuing medical education provision in Singapore. PMID- 10672414 TI - Dimerization kinetics of violamycin BI anthracycline--the influence of ionic strength. AB - Violamycin BI is an anthracycline derivative with two sugars hanging on, each of them carries one positive charge. It dimerizes under conditions, which depend on the concentration of the antibiotic, pH and the ionic strength of the solution. By keeping a constant pH in a phosphate-EDTA buffer, the rate constants of violamycin BI dimerization were determined at various ionic strengths by temperature jump method. The dimerization constant Kd, resulting from the ratio of these rate constants, confirmed the values obtained spectrophotometrically in this study or elsewhere. The influence of ionic strength (0.02-0.2 M) on the rate constant values suggested to us some speculations on the reaction mechanism of the dimerization, in which, the specific mutual orientation of the monomers in the encounter, and perhaps a specific conformation of their side groups is required before a stabilizing action of the binding forces sets in. PMID- 10672415 TI - Pathways involved in trifluoperazine-, dibucaine- and praziquantel-induced hemolysis. AB - This work elucidates differences in the hemolytic pathway developed by the antipsychotic trifluoperazine (TFP), the local anesthetic dibucaine (DBC) and the antihelminthic praziquantel (PZQ). Their partition coefficients (P) were measured at pH 7.4 between n-octanol, microsomes, liposomes, erythrocyte ghosts and n octanol/water. The effective drug:lipid molar ratios for the onset of membrane solubilization (ReSAT) and complete hemolysis (ReSOL) were calculated from the experimental P values and compared with a classical surface-active compound treatment Lichtenberg, D. Biochim. Biophys. Acta 821 (1985) 470-478[. The contribution of charged/uncharged forms of TFP and DBC for the hemolytic activity was also analyzed. In all cases the hemolytic phenomena could be related to the monomeric drug insertion into the membrane. Only for TFP at isosmotic condition lysis occurs at concentrations beyond the CMC of the drug, indicating that micellization facilitates TFP hemolytic effect, while DBC and PZQ reach a real membrane saturation at their monomeric form. PMID- 10672416 TI - A spectroscopic study of the molecular interactions of harmane with pyrimidine and other diazines. AB - FTIR, UV-vis, steady state and time-resolved fluorescence measurements show that harmane (1-methyl-9H-pyrido/3,4-b/indole) interacts with pyrimidine and its isomers pyrazine and pyridazine in its ground and lowest singlet states. The mechanisms of interaction are dependent on both the structure of the diazine and the nature of the solvent. Thus, in a low polar solvent such as toluene, harmane forms ground state 1:1 hydrogen-bonded complexes with all the diazines. These complexes quench the fluorescence of harmane and diminish its fluorescence lifetime. Conversely, in buffered (pH 8.7) aqueous solutions, pyrimidine behaves differently from the other diazines. Thus, whereas pyrimidine only interacts with harmane in its ground state, pyrazine and pyridazine also interact in the excited state. The harmane-pyrimidine ground state interaction is an entropic controlled process. Therefore, we propose the formation of pi-pi stacked 1:1 complexes between these substrates. Association constants for the different types of complexes and quenching parameters are reported. PMID- 10672417 TI - In vitro effects of sodium fluoride and sodium dichromate on dynamic properties of human erythrocyte membrane. AB - Sodium fluoride (NaF) and sodium dichromate (Na2Cr2O7) are two different toxic compounds which are used as a dental caries prophylactic and as an oxidising agent in various industrial areas, respectively. However, accidental fluoride and chromate poisoning is not a rare occurrence, even death may result from cardiac or respiratory failure. In the present work, alterations produced by NaF, Na2Cr2O7 and temperature changes in the molecular dynamics of the human erythrocyte membrane were studied, in vitro, by the spin-labelling ESR technique. Human intact erythrocyte cells spin labelled with 5- and 16-doxyl stearic acids (5-DSA and 16-DSA) and treated with 40 microM NaF and 5 microM Na2Cr2O7 at 37 degrees C were used to quantify membrane fluidity. This was performed by measuring the changes in the order parameter (S), correlation time (tau) and phase transition temperature using recorded electron spin resonance (ESR) spectra. Experimental results show that 5 microM Na2Cr2O7 and 40 microM NaF do not produce any significant effects on the order parameter of 5-DSA spin label while they cause appreciable changes in the correlation time of the same label. As for 16-DSA, while Na2Cr2O7 does not produce any measurable effect on the order parameter of this label, NaF does in a certain extent. Although weak, the effects of both compounds on the correlation time of 16-DSA are found to be well above the experimental error limits. Change in temperature was observed to alter significantly S and tau parameters which show biphasic character in the temperature range of 5-50 degrees C. Activation energies of the hydrocarbon chains above and below transition temperatures were also determined for untreated and NaF or Na2Cr2O7 treated erythrocyte cells and the effect of NaF and Na2Cr2O7 on these energies and transition temperatures were discussed. PMID- 10672418 TI - Self-assembled complexes of oligopeptides and metalloporphyrins: measurements of the reorganization and electronic interaction energies for photoinduced electron transfer reactions. AB - Cationic porphyrins form ground state electrostatically associated complexes with anionic oligo-electrolytes such as those formed by a series of glutamic acid (E) residues. Temperature dependencies were measured of the rate constants for intra complex electron transfer to the triplet state of Pd(II)TMPyP4+ from a tyrosine (tyr, Y) or tryptophan (trp, W) moiety connected to a glutamic acid tetramer. In complexes such as YE4, E2YE2, YE4G10E (G, glycine), and WE4 these data were used to estimate the reorganization energy (lambda) and electronic interaction energy (HDA) relevant to the process. For all tyr-peptide complexes, lambda values were found to be large (lambda approximately 1.60 +/- 0.06 eV), reflecting a relatively high medium polarity in the vicinity of tyr residues. It further indicates that the tyr residues in all oligo-peptides are exposed to the aqueous medium in a similar way irrespective of the position of the aromatic moiety in the peptide chain. A significantly lower lambda value (lambda = 1.08 eV) was derived for the tryptophan-containing peptide complex, indicating a relatively higher hydrophobic character of trp compared to tyr. The electronic coupling matrix elements (HDA) derived for tyr-peptide complexes (5.1 meV for YE4, 5.4 meV for YE4G10E and 7.5 meV for E2YE2) were larger than that found for WE4 (1.1 meV). Molecular dynamics calculations were employed to obtain structural features of the porphyrin-peptide complexes. These showed average distances between the center of mass (COM) of the porphyrin ring and the center of mass of the amino acid aromatic ring of 816 +/- 140 pm (YE4), 800 +/- 80 pm (E2YE2), 900 +/- 130 pm (YE4G10E) and 970 +/- 160 pm (WE4). The molecular dynamics calculations were shown to be in good agreement with the experimentally determined electronic interaction energies, strongly suggesting that HDA is primarily responsible for the dependence of the electron-transfer rate constant (KET) on the donor-acceptor separation distance and relative orientation. The higher HDA (7.55 meV) derived for tyr incorporated into the middle of the peptide backbone (E2YE2) was presumed to be associated with a higher degree of orbital overlap due to a more favorable ring-ring orientation. Overlap parameters (beta derived for all peptide-porphyrin complexes were similar (approximately 0.95 +/- 0.06 A-1), being in good agreement with most literature values for similar systems. Finally, the intra-complex electron-transfer ratio (ktrp/ktyr) derived from flash photolysis experiments and the corresponding ratio derived from Marcus' theory combined with experimental data from the temperature-dependence investigations and electrochemical measurements were found to be in excellent agreement. This same consistency was found for the couple E4Y and E2YE2. The empirical expression (Moser and Dutton) governing the intraprotein electron-transfer rate constant in native systems combined with our experimental data (kET, lambda, delta G0) yielded tunneling pathway distances in excellent agreement with those arising from the molecular modeling studies. The exception was for the long peptide YE4G10E, for which the Quenched Molecular Dynamic (QMD) sampling technique was complicated and is probably inadequate. PMID- 10672419 TI - Complexes produced by associated forms of porphyrin bound with hydrophobic hydrophilic copolymer and transition metal ions. AB - Properties of protonated dimeric forms of meso-tetraphenylporphine (TPP) and meso tetra(p-aminophenyl)porphine (TAPP) bound with copolymer and also complexes produced by associated TAPP bound with copolymer, Mn2+, and Fe3+ are investigated by absorption, luminescence, and Raman spectroscopy. According to absorption spectra of protonated dimers of TPP, three dimeric forms of the porphyrin are observed in the ground state. However, selective excitation of these forms according to the fluorescence spectra reveals only two dimeric forms in the excited state. In contrast, similar selective excitation of TAPP bound with copolymer in aqueous-dioxane solution results in weak changes in the fluorescence spectra, nevertheless, there is strong interaction between porphyrin and macromolecular carboxyl groups in the ground state. In the case of the formation of the complexes between associated TAPP bound with copolymer, Mn2+ and Fe3+, a new band in the near IR region with a maximum at 840 nm is built up in the fluorescence spectrum. However, this near IR emission is completely quenched when new strong vibrational bands at approximately 1800 and 1900 cm-1 are revealed in the resonance Raman spectra of the complexes. The observed effects are explained in terms of direct participation of water molecules involved in the water porphyrin dimeric complex in the processes of transformation of excitation energy. The involvement of water in this dimeric complex can lead to redistribution of flows of the energy degradation when transition metal ions play a role of the agent which enhances the trapping properties of the porphyrin-metal ions complexes. PMID- 10672420 TI - Oscillations and bistability predicted by a model for a cyclical bienzymatic system involving the regulated isocitrate dehydrogenase reaction. AB - We analyze the dynamics of a bienzymatic system consisting of isocitrate dehydrogenase (IDH, EC. 1.1.1.42), which transforms NADP+ into NADPH, and of diaphorase (DIA, EC 1.8.1.4), which catalyzes the reverse reaction. Experimental evidence as well as a theoretical model showed the possibility of a coexistence between two stable steady states in this reaction system G.M. Guidi et al. Biophys. J. 74 (1998) 1229-1240[, owing to the regulatory properties of IDH. Here we extend this analysis by considering the behavior of the model proposed for the IDH-DIA bienzymatic system in conditions where the system is open to an influx of its substrates isocitrate and NADP+ and to an efflux of all metabolic species. The analysis indicates that in addition to different modes of bistability (including mushrooms and isolas), sustained oscillations can be observed in such conditions. These results point to the isocitrate dehydrogenase reaction coupled to diaphorase as a suitable candidate for further experimental and theoretical studies of bistability and oscillations in biochemical systems. The results obtained in this particular bienzymatic system bear on other enzymatic systems possessing a cyclical nature, which are known to play significant roles in a variety of metabolic and cellular regulatory processes. PMID- 10672421 TI - Pneumocephalus associated with aqueductal stenosis: three-dimensional computed tomographic demonstration of skull-base defects. AB - Ventriculoperitoneal (VP) shunt placement in patients with aqueductal stenosis has recently been reported as a cause of pneumocephalus. We report on a patient with pneumocephalus associated with aqueductal stenosis treated by VP shunting. A 29-year-old woman who had undergone a shunt operation for aqueductal stenosis 7 years previously sustained a whiplash injury in a minor traffic accident. Computed tomography (CT) revealed massive subdural pneumocephalus, and three dimensional reconstructions of CT images clearly demonstrated defects in the skull base overlying the ethmoid sinuses. Both endoscopic III ventriculostomy and placement of external ventricular drainage were came free of symptoms and rhinorrhea ceased. Three-dimensionally reconstructed CT images were useful in detecting the extent of the patient's skull base defect. III Ventriculostomy was not effective in this case. Direct closure of the skull base by craniotomy was not necessary, and a programmable valve system was effective in preventing recurrence of either pneumocephalus or rhinorrhea. PMID- 10672422 TI - The treatment of Candida albicans shunt infections. AB - Cerebrospinal fluid shunting procedures are performed for the treatment of hydrocephalus. Infection of ventriculoperitoneal shunts may create significant clinical management issues in these patients. The majority of these infections are bacterial, but occasionally a Candida albicans shunt infection may occur. We report two patients who acquired Candida albicans shunt infection and discuss their clinical presentation, management, and successful outcome. The treatment with or without removal of the shunt and the correct dosage and route of administration of the antifungal agents is not well documented. The dilemma of treatment of Candida albicans shunt infections in these patients and review of the limited literature on this subject are the subjects of this report. PMID- 10672423 TI - The management of desmoplastic neuroepithelial tumours in childhood. AB - The authors report on the clinicopathological aspects of and management strategies for the group of rare, large hemispheric childhood tumours recently classified as desmoplastic infantile ganglioglioma (DIGG), desmoplastic astrocytoma of infancy (DACI) and pleomorphic xanthoastrocytoma (PXA). Between 1985 and 1997, ten children (4 with DACIs, 4 with DIGGs and 2 with PXAs) with a median age of 9.5 months were operated on. All these patients had complete surgical resections, with two having a preoperative biopsy. This led to an erroneous diagnosis in both cases of malignant grade 4 astrocytoma. As a result, one patient had preoperative chemotherapy with no effect. There was one perioperative death. Histology revealed heterogeneous tumours with malignant looking areas in 8 of the specimens. None of the patients has had any postoperative adjuvant treatment. All surviving patients are alive at follow-up (median 4.2 years). Despite their often malignant appearance, these tumours have an excellent prognosis, but they can present formidable surgical challenges when they occur in very young age patients. We believe that surgical excision can offer a cure and that adjuvant treatment is not necessary. Finally, biopsy is of little value and may even lead to an erroneous diagnosis and subsequent mismanagement. PMID- 10672424 TI - Management of medulloblastoma and ependymoma in infants: a single-institution long-term retrospective report. AB - To reduce the sequelae from CNS irradiation (RT), 16 children younger than 3 years with medulloblastoma-PNET (13 cases) and ependymoma (3 cases) were treated between 1987-1993 according to different postsurgical chemotherapy (CT) programs. None of these patients presented with metastases. Eleven patients were rendered disease-free by surgery, while 5 had residual tumor. Adjuvant therapy depended on patients' age, postsurgical status and parents' consent to radiotherapy (RT). Nine of the 16 infants remained alive in continuous complete remission from the first neoplasm (median follow-up 7 years). Three of them had been treated with CT alone and 6 with combined CT + RT (posterior fossa 4, whole CNS 2). Seven patients relapsed a median of 13 months after diagnosis, and all 7 of them died of their disease. Despite the omission of RT in 6 of the 16 patients and administration of only focal RT in 8 of the 16, the outcome of this series was satisfactory. Local failure (in 5/7 patients) was the major problem, despite the high dose of RT used in 2 of these 5. In 4 of 6 evaluable children school performance was satisfactory. One child in whom the entire CNS was irradiated developed glioblastoma multiforme 120 months after the first diagnosis of medulloblastoma. PMID- 10672425 TI - Traumatic brain stem lesions in children. AB - Glasgow Coma Scale (GCS) scores on admission may be predictors of outcome in patients with brain injuries. This study correlated the outcomes of children with traumatic brain stem lesions with their initial GCS scores and morphological patterns of injury as shown on computed tomography (CT) or magnetic resonance (MR) imaging. During the last 16 years, we have treated 1,108 children with brain injuries. The entire series included only 21 (1.9%) children who had clinical signs of brain stem lesions with morphological correlates on CT or MR imaging. Clinical findings were assessed according to the GCS and compared with scores on the Glasgow Outcome Scale (GOS). Of these 21 children, 16 (76%) had morphological lesions seen on CT scans. In 5 (24%) of the children only the MR images revealed brain stem lesions and their CT scans were negative. Generalized severe brain swelling was present in 6 cases (28%). There was a significant difference in GOS scores between patients with initial GCS scores of 3 and 4 and those with GCS scores between 5 and 7 (P < 0.02). Children with intracranial pressure higher than 40 mmHg had poorer outcomes than patients whose intracranial pressure was lower, but the differences were not significant. Outcome did not correlate significantly with morphological patterns of injury or the presence of extracranial injuries. The GCS is a reliable indicator of severity of injury and of outcome in children with brain stem injuries. MR imaging was more sensitive than CT in detecting brain stem lesions. PMID- 10672426 TI - Acute neuroradiologic findings in young children with inflicted or noninflicted traumatic brain injury. AB - Acute CT/MRI findings were examined in a prospective, longitudinal study of 60 children 0-6 years of age hospitalized for moderate to severe traumatic brain injury (TBI). TBI was categorized as either inflicted (n = 31) or noninflicted (n = 29). Glasgow Coma Scale scores and perinatal history were comparable in both groups. Acute CT/MRI studies were visually inspected by a radiologist blind to group membership. Compared with the noninflicted TBI group, the inflicted TBI group had significantly elevated rates of subdural interhemispheric and convexity hemorrhages as well as signs of pre-existing brain abnormality, including cerebral atrophy, subdural hygroma, and ex vacuo ventriculomegaly. Intraparenchymal hemorrhage, shear injury, and skull fractures were more frequent after non-inflicted TBI. Subarachnoid hemorrhage and infarct/edema occurred with comparable frequency in both groups. Characteristic acute neuroimaging findings of inflicted TBI included multiple extraaxial hemorrhages in addition to the mild atrophy, subdural hygromas, and ventriculomegaly that suggest prior brain abnormality. PMID- 10672427 TI - Stormy onset of benign childhood epilepsy with occipital paroxysmal discharges. AB - We studied six children with ages ranging from 4 to 10 years who were affected by childhood epilepsy with occipital paroxysms and presented after a stormy onset with prolonged loss of consciousness for 6-14 h. In all these patients, seizures were preceded by visual symptoms in the form of colored circular disks. A CT scan was performed immediately after the onset of symptoms and was normal in all patients. Routine laboratory and cerebrospinal fluid examinations were normal in all patients. The interictal EEG was characterized by continuous or subcontinuous occipital spike wave discharges, which disappeared after the patients' eyes opened. We carried out a 7-year follow-up of all these patients. Only two patients were treated with antiepileptic drugs. The therapy (phenobarbital, clobazam) in the two patients did not induce changes in the EEG pattern. The first did not suffer any further seizures. The second patient had two more seizures (at 8 and 18 months from the onset) with phosphenes, confusional state, and involuntary movements followed by loss of consciousness. Among the other four patients, who did not receive any treatment, only one had any other seizures. The stormy onset of the syndrome described in our six patients emphasizes the extreme variability in the presentation of this type of childhood epilepsy. Our follow-up confirms the good prognosis of this epilepsy even when it has a stormy onset. PMID- 10672428 TI - Brain death in children: clinical, neurophysiological and radioisotopic angiography findings in 125 patients. AB - The objective of this study was to determine the main clinical, neurophysiological and angiographic findings in brain death (BD) in children seen at the Instituto Nacional de Pediatria, a third-level facility in Mexico City, between 1991 and 1996. The following variables were retrospectively analyzed: sex, age, etiology, associated morbidity, duration of stay in hospital, and the results of two of three confirmatory studies (electroencephalogram, evoked potentials, radioisotopic angiography). In all, 125 patients were studied 78 male, median age 2 years (range: 18 days to 17 years)[. The most frequent etiology was infection (34%); 57% of the children developed associated morbidity. In 111 of 122 patients electrocerebral silence was observed; 100 of 107 had brain stem and somatosensory evoked potentials affording conclusive evidence of BD; and 83 of 90 patients had a positive radioisotopic angiography indicating BD. In 76 patients all three confirmatory studies were performed: for 15 there was at least one false-negative test result. Our age cohort showed a predominance of children less than 2 years old. BD etiologies in developing countries differ from those reported in developed countries. PMID- 10672429 TI - Period shift induction by intermittent stimulation in a Drosophila model of PER protein oscillations. AB - PER protein circadian oscillations in Drosophila have been described by Goldbeter according to a five-dimensional model that includes the possibility of genetic mutation described by changing one parameter, the maximum degradation rate of the PER protein. Assuming that, in a mutant Drosophila this parameter is unreachable, we modify another parameter, the translation rate between the mRNA and the nonphosphorylated form of PER protein, by periodic intermittent activation or inhibition. We show how such a modification, simulated in the model by a periodic, on/off, piecewise constant stimulation (which increases or decreases this parameter) allows the entrainment of oscillations exactly at, or close to, a desired period. In a different context, this suggests that some diseases may be corrected using pharmacological agents according to specific periodic delivery schedules. PMID- 10672430 TI - Pineal influence on annual nuclear volume changes in ventromedial hypothalamic nucleus (VMH) neurons of the male Wistar rat. AB - The ventromedial hypothalamic nucleus (VMH) regulates various autonomic, endocrine, and behavioral activities. These activities show annual changes, and the pineal gland is involved in their adjustment to environmental cues. Therefore, this study investigated whether the VMH belongs to the effector structures of the pineal gland. To abolish the rhythmic melatonin release, male Wistar rats were subjected to pinealectomy (PX) or ganglionectomy (sympathetic denervation of the pineal gland, GX) regularly at the beginning of any of the four seasons. Brains from animals of PX-, GX-, and sham-operated control groups were prepared 3 months later for measurement of the nuclear volume, which changes according to the general gene activity. At each of the four seasons, 2000 nuclei of VMH neurons stemming from 18 animals per group were measured to obtain both seasonal daily mean values and annual mean values, respectively, as well as to calculate annual curves of the nuclear volume using empirical regression and locally adjusted polynomial approximation. The major findings are the following. First, inactivation of the pineal function influences the nuclear activity of VMH neurons, (2) PX and GX mainly depress the nuclear activity, indicating that the pineal influence on the VMH may predominantly be a stimulatory one. Third, size and direction of the changes caused by PX and GX vary in a seasonally dependent manner. Fourth, the annual rhythm of the nuclear activity of the VMH is modified by PX and GX. To explain how the pineal effects on the VMH may be mediated, a possible inhibitory influence of the suprachiasmatic nucleus (SCN), which has been activated in the same animals following both PX and GX, is discussed. In conclusion, the results confirm that the nuclear activity of VMH neurons underlies pineal influences. This also indicates an involvement of the pineal gland in many VMH-regulated functions. PMID- 10672431 TI - Quantitative tests of a dual circalunidian clock model for tidal rhythmicity in the sand beach isopod Cirolana cookii. AB - In constant conditions (constant darkness DD[, 20 degrees C), the sand beach isopod Cirolana cookii exhibits spontaneous rhythmic swimming activity with an average free-running period of 12.5 h. The rhythms are seen as temporal adaptations to a complex interidal environment. These results support a dual circalunidian clock model for tidal rhythms in which two components of the rhythm have characteristic periods and active phase lengths and are hypothesized to be controlled by separate circalunidian clocks. A quantitative model successfully simulates many of the properties of endogenous swimming rhythms of C. cookii, including free-running behavior, entrainment, and phase-response curves (PRCs). PMID- 10672432 TI - Evidence of circadian rhythm of electric discharge in Eigenmannia virescens system. AB - The gymnotid electric fish, Eigenmannia virescens, exhibits electric discharge rhythmicity both in alternate light-dark (LD; 12 h light, 12 h dark [LD 12:12]) and in constant dark (DD) conditions. It suggests that the electric discharge rhythm is under control of the circadian clock. The free-running periods (FRPs) of electric discharge rhythms at 21 degrees C in DD are greater than, but close to, 24 h. The maximum of the electric discharge in the Eigenmannia system peaks approximately at circadian time 6 (CT6) in the middle of the subjective day. The circadian oscillator in the system is temperature compensated. This original report reveals the relationship between electric discharge activity and the circadian pacemaker in Eigenmannia and provides an alternative system to investigate circadian rhythms in vertebrates. PMID- 10672433 TI - The sleep of healthy people--a diary study. AB - To provide baseline data for various research studies at the University of Pittsburgh over a 10-year period, 266 healthy subjects (144 male, 122 female, aged 20-50 years) meeting certain criteria each completed a 14-night sleep diary. For each night, the diary allowed the subjective measurement of bedtime, wake time, time in bed (TIB), sleep efficiency, number of minutes of wake after sleep onset (WASO), alertness on awakening, and percentage of morning needing an alarm (or a person functioning as one). Weeknight versus weekend night differences in TIB (TIBdiff), weekday altertness, and reliance on alarms were examined as possible indicators of sleep debt. In addition, general descriptive data were tabulated. On average, bedtimes were at 23:48 and wake times at 07:23, yielding a mean TIB of 7 hours 35 minutes. As expected, bedtimes and wake times were later on weekend nights than on weeknights. Bedtimes were 26 minutes later, wake times 53 minutes later, yielding a mean weekend TIB increase of 27 minutes. Overall, subjects perceived their sleep latency to be 10.5 minutes, reported an average of one awakening during the night (with an average of 6.4 minutes of WASO), had a diary sleep efficiency of 96.3%, and awoke with an alterness rating of 69.5%. These variables differed little between weeknight and weekend nights. Subjects used an alarm (or a person functioning as an alarm) on 60.9% nights overall, 68.3% on weeknights, 42.5% on weekends. When TIBdiff was used as an estimate of sleep debt (comparing subjects with TIBdiff > 75 minutes with those with a TIBdiff < 30 minutes), the group with more "catch-up sleep" on weekends had shorter weeknight TIB durations (by about 24 minutes) and relied more on an alarm for weekday waking (by about 22%), indicating the possible utility of these variables as sleep debt indices. PMID- 10672434 TI - Day-night pattern in accidental exposures to blood-borne pathogens among medical students and residents. AB - The purpose of this study was to determine whether the occurrence of accidental blood-borne pathogen exposure incidents in medical students and residents in training varies during the 24 h. A retrospective review of reported exposures was conducted in a large urban teaching institution--the University of Texas Health Science Center in Houston--between November 1993 and July 1998. Professional level (year of student or level of resident), time of exposure, means/route of exposure (needle stick, laceration, or splash), and type of medical service were recorded. Analysis of the clock time of the 745 reported blood-borne pathogen exposures showed they occurred more frequently during the day than night. Over the nearly 5-year span, 531 incidents took place between 06:00 and 17:59 in comparison to only 214 between 18:00 and 05:59. To account for the day-night difference in medical student and resident hospital staffing, the data were reexpressed as exposure rates, that is, in terms of the number of events per hour per 1000 medical students and residents. Based on the total number of reported exposures over the almost 5-year span of data collection, the average rate was 40 accidents per hour per 1000 doctors in training during the 12 h daytime span (6:00-17:59). It was 50% greater at night (18:00-05:59), with 60 incidents per hour per 1000 doctors in training. The day-night difference in rate of exposures was statistically significant (p < .04). The relative risk ratio for residents and students when working during the day shift compared to working the night shift was 0.67. This means that doctors in training are at a 1.50 higher risk of sustaining a blood-borne pathogen exposure when working nights than when working days. PMID- 10672435 TI - Effect of melatonin on sleep quality of COPD intensive care patients: a pilot study. AB - Sleep deprivation is extremely common in the intensive care unit (ICU), and this lack of sleep is associated with low melatonin secretion. The objective of the current study was to explore the effect of exogenous melatonin administration on sleep quality in patients hospitalized in the pulmonary intensive care unit (ICU). We performed a double-blind, placebo-controlled study in the pulmonary ICU of a tertiary care hospital. Eight adult patients hospitalized in the pulmonary ICU with respiratory failure caused by exacerbation of chronic obstructive pulmonary disease (COPD) or with pneumonia were studied. Patients received either 3 mg of controlled-release melatonin or a placebo at 22:00, and sleep quality was evaluated by wrist actigraphy. Treatment with controlled-release melatonin dramatically improved both the duration and quality of sleep in this group of patients. Our results suggest that melatonin administration to patients in intensive care units may be indicated as a treatment for sleep induction and resynchronization of the "biologic clock." This treatment may also help in the prevention of the "ICU syndrome" and accelerate the healing process. PMID- 10672436 TI - Algorithm for application of Fourier analysis for biorhythmic baselines of pharmacodynamic indirect response models. AB - The change of an indirect pharmacological response R(t) can be described by a periodic time-dependent production rate kin(t) and a first-order loss constant kout. If kin(t) follows some biological rhythm (e.g., circadian), then the response R(t) also displays a periodic behavior. A new approach for describing the input function in indirect response models with biorhythmic baselines of physiologic substances is introduced. The present approach uses the baseline (placebo) response Rb(t) to recover the equation for kin(t). Fourier analysis provides an approximate equation for Rb(t) that consists of terms (usually two or three) of the Fourier series (harmonics) that contribute most to the overall sum. The model differential equation is solved backward for kin(t), yielding the equation involving Rb(t). A computer program was developed to perform the square L2-norm approximation technique. Fourier analysis was also performed based on nonlinear regression. Cortisol suppression after inhalation of fluticasone propionate (FP) was modeled based on the inhibition of the secretion rate kin(t) using ADAPT II. The pharmacodynamic parameters kout and IC50 were estimated from the model equation with kin(t) derived by the new approach. The proposed method of describing the input function needs no assumption about the behavior of kin(t), is as efficient as methods used previously, and is more flexible in describing the baseline data than the nonlinear regression method. PMID- 10672437 TI - Chronic fatigue syndrome beginning suddenly occurs seasonally over the year. AB - The fact that many patients with chronic fatigue syndrome (CFS) have an infectious like sudden onset to their illness has led to the hypothesis that CFS is a medical illness. If CFS were, on the other hand, a psychiatric disorder related to symptom amplification, one would expect illness onset to occur randomly over the calendar year. This study tested that hypothesis with 69 CFS patients whose illness was on the more severe side of the illness spectrum; all patients reported sudden illness onset with the full syndrome of sore throat, fatigue/malaise, and diffuse achiness developing over no longer than a 2-day period. Date of illness onset was distinctly nonrandom. It peaked from November through January and was at its lowest from April through May. These data support the hypothesis that an infectious illness can trigger the onset of CFS. PMID- 10672438 TI - Examination of the transcriptional specificity of an enological yeast. A pilot experiment on the chromosome-III right arm. AB - The adaptation of yeasts to industrial environments is thought to be largely dependent on gene-expression specificity. To assess the transcriptional specificity of an enological strain, we performed a pilot experiment and examined the transcript level of 99 ORFs of the chromosome-III right arm with two strains, an enological-derived strain and a laboratory strain, grown under three different physiological conditions: respiration, standard alcoholic fermentation and enological alcoholic fermentation. The use of 99 single ORF-derived probes led to the detection of 49 transcripts, most of which were present at low levels and were not regulated. Ethanol respiration induced transcripts, in a similar manner with both strains. While standard alcoholic fermentation led to only minor regulations, the enological fermentation conditions triggered the expression of different genes. In addition, a specific transcriptional response to these conditions was observed with the enological-derived strain. The known or predicted functions of several genes induced under enological conditions is related to either alcoholic fermentation or stress, suggesting that their specific induction could reflect adaptation of the strain to the enological environment. Our data suggest that systematic transcriptional studies are an effective way to assess the molecular basis of yeast adaptation to industrial environments. PMID- 10672439 TI - The cyanobacterial origin and vertical transmission of the plastid tRNA(Leu) group-I intron. AB - We have surveyed the distribution and reconstructed the phylogeny of the group-I intron that is positioned in the anticodon loop of the tRNA(Leu) gene in cyanobacteria and several plastid genomes. Southern-blot and PCR analyses showed that the tRNA(Leu) intron is found in all 330 land plants that were examined. The intron was also found, and sequenced, in all but one of nine charophycean algae examined. Conversely, PCR analyses showed that the tRNA(Leu) group-I intron is absent from the red, cryptophyte and haptophyte algae, although it is present in three members of the heterokont lineage. Phylogenetic analyses of the intron indicate that it was present in the cyanobacterial ancestor of the three primary plastid lineages, the Rhodophyta, Chlorophyta, and Glaucocystophyta. Its present day distribution in plastids is consistent with a history of strictly vertical transmission, with no losses in land plants, several losses among green algae, and nearly pervasive loss in the Rhodophyta and its secondary derivatives. PMID- 10672440 TI - Recombinant mitochondrial DNA molecules suggest a template switching ability for group-II-intron reverse transcriptase. AB - Degenerative processes in the filamentous fungus Podospora anserina are strongly correlated with the instability of the mitochondrial genome. Among the sources of instability is the mobile group-II intron COX1-i1, also called intron alpha, which encodes a protein with a reverse transcriptase activity. In this paper we characterize, through PCR experiments, mitochondrial recombinant DNA molecules joining the 5' end of intron alpha to the 3' end of tRNA sequences including the CCA motif. The structure of these junctions led us to propose that they were most probably initiated by a RNA template switching of the reverse transcriptase encoded in COX1-i1. This activity might be involved in a number of mitochondrial rearrangements occurring in degenerative syndromes and in some long-lived mutants. PMID- 10672441 TI - Phylogenetic analysis of diatom coxI genes and implications of a fluctuating GC content on mitochondrial genetic code evolution. AB - In order to address the relationships among diatom groups and to investigate possible changes in their mitochondrial (mt) genetic codes, we have analyzed a 1.1-kb region of the cytochrome c oxidase subunit I (coxI) gene from eight diverse diatom species. A phylogenetic analysis of these coxI sequences including representative species of the Phaeophyta, Xanthophyta, Eustigmatophyta and Haptophyta showed that the diatoms (Bacillariophyta) formed a well-supported monophyletic group. Of the eight species investigated, four have been classified together as radial centric diatoms based on morphology. However, in our coxI tree, the two radial centrics belonging to the order Thlassiosirales (Skeletonema costatum and Thalassiosira nordenskioldii) were placed as the sister group to the multipolar centric diatoms, while the other two radial centrics (Melosira ambigua and Rhizosolenia setigera) were in another clade. Also, in two species of the Tharassiosirales we found UGA codons that occur at conserved tryptophan (Trp) sites in the coxI sequences, strongly indicating that UGA codes for Trp in these diatoms. No evidence of a deviant genetic code was detected in the other analyzed diatom species. There was no apparent relationship between the nucleotide third position GC content of mtDNA (based on the sequenced coxI region) and the presence of a deviant genetic code. PMID- 10672442 TI - Variable number of tandem repeat loci in the mitochondrial genomes of beets. AB - We found four unrelated tandem repeat loci (TR1, TR2, TR3 and TR4) in the mitochondrial genomes of beets, with the TR1 locus embedded within a three membered family of recombining repeat sequences (the rrn26-repeat). TR1 is composed of an array of 32-bp tandem repeats, the number of which varies from 2 to 13 among the seven beet genotypes examined. It is interesting to note that TR1 has 7-bp direct repeats flanking the array, which may be involved in the generation of the tandem repeat array. Such striking features are shared by the remaining TR loci, and this is thus the first description of minisatellite nucleotide sequences from a higher-plant mitochondrial genome. PMID- 10672443 TI - Kalilo plasmids are a family of four distinct members with individual global distributions across species. AB - Kalilo is a linear 9-kb plasmid, isolated originally from Hawaiian strains of the heterothallic fungus Neurospora intermedia. Its properties include terminal inverted repeats, two ORFs coding for a presumptive DNA and an RNA polymerase, and the ability to cause senescence in its original host and in the closely related species Neurospora crassa. We have examined natural isolates alleged to contain plasmids homologous to kalilo. Most of these isolates do in fact contain plasmids with so close an identity to kalilo as to be certain relatives. We found a new case of kalilo in Neurospora tetrasperma from Moorea-Tahiti, and a new case of LA-kalilo (previously found only in N. tetrasperma) in N. crassa from Haiti. A previously unreported, substantially shorter, kalilo variant has been found in three geographically separate isolates of the heterothallic species Neurospora discreta. Therefore, if the previously reported kalilo variant from the genus Gelasinospora is included, in all there are four members of the kalilo plasmid family. The main differences between these plasmids are in the terminal inverted repeats (TIRs). The phylogeny of the TIR sequences is largely congruent with that of nuclear DNA in the species in which they are found, suggesting that the plasmids are related by vertical descent throughout the evolution of these species. However, there are two cases of a plasmid found in a heterothallic and a pseudohomothallic species in the same global area; these cases might have arisen from more recent horizontal transmission or introgression. PMID- 10672444 TI - Transcripts and sequence elements suggest differential promoter usage within the ycf3-psaAB gene cluster on mustard (Sinapis alba L.) chloroplast DNA. AB - The mustard chloroplast DNA region spanning the ycf3 gene and part of the psaAB operon was investigated. The ycf3 gene reveals two class-II introns that are removed during processing to give a mature 0.7-kb transcript, but no RNA editing seems to be involved. RNase protection and RT-PCR experiments suggest cotranscription of ycf3 with the downstream psaA gene, possibly from a NEP promoter upstream of ycf3, whereas distinct ycf3 and psaA transcripts are each initiated from PEP promoters. This situation is reminiscent of that for the trnK psbA gene region. The implications for light-regulated versus light-independent expression of photosystem core-protein genes are discussed. PMID- 10672445 TI - Maternal inheritance of chloroplasts in the horsetail Equisetum variegatum (Schleich.). AB - Reliable data concerning the transmission of chloroplasts in the Pteridophyta are needed both for phylogenies based on chloroplast DNA (cpDNA) sequences and in order to study the evolution of this trait in conjunction with the evolution of the life cycle and the sexual reproduction of land plants. For the first time, this paper describes organelle transmission in the division Sphenophyta, represented by the extant genus Equisetum. By following the fate of polymorphic cpDNA during three intraspecific reciprocal crosses we found no trace of paternal transmission in Equisetum variegatum. The seemingly strict maternal transmission of cpDNA in this species suggests that uniparental chloroplast inheritance preceded the evolution of heterospory in the seed-plant lineage. PMID- 10672446 TI - Molecular characterization of a PDI-related gene prpA in Aspergillus niger var. awamori. AB - A gene (prpA) homologous to the protein disulfide isomerase gene was isolated from Aspergillus niger by Southern hybridization using the pdi1 gene isolated from Trichoderma reesei as a DNA probe. The corresponding cDNA of the prpA gene has also been isolated from an A. niger var. awamori cDNA library. The prpA gene does not belong to any currently recognized family of protein disulfide isomerases since it contains only a single conserved thioredoxin domain at the N terminus of the protein. The C-terminal two-thirds of the protein has no homology to any known proteins in the database. The PRPA protein contains an ER retention signal (HDEL) at its C-terminal end suggesting that it is located in the ER. Southern hybridization at high stringency showed that it was present as a single copy in the genome. Northern hybridization indicated that the transcript level of the prpA gene was higher if the cells were secreting a heterologous protein, bovine prochymosin. However, over-expression of the prpA gene from a multicopy integrated vector had little effect on chymosin secretion. A strain containing a deletion of the prpA gene was viable. However, deletion of the prpA gene appeared to cause a reduction of bovine chymosin production. PMID- 10672448 TI - The 20th century--the frame that we leave. PMID- 10672447 TI - Cytochrome P450 oxidoreductase gene and its differentially terminated cDNAs from the white rot fungus Phanerochaete chrysosporium. AB - The white rot fungus Phanerochaete chrysosporium metabolizes a range of xenobiotics via P450 mono-oxygenation, particularly under peroxidase-suppressing culture conditions. Here we report the cloning and analysis of the gene from this fungus for the cytochrome P450 oxidoreductase (CPR) and its differentially terminated cDNAs. Using a PCR-based approach with degenerate primers, a 285-bp genomic fragment was isolated from the two widely studied strains BKM-F 1767 and ME 446, and was identified as a CPR gene segment based on sequence comparison with the database. A clone containing the full-length CPR gene was isolated from a BKM-F 1767 genomic library using the PCR-generated segment as a probe, and the 3937-bp insert was sequenced by gene walking. Based on the detection of conserved CPR motifs, a coding region of 2381 bp was identified with a 991-bp segment 5' to the putative ATG start codon. Two cDNAs with differentially terminated transcripts were isolated and sequenced. Comparison of the gene and the cDNA sequences confirmed the presence of three introns (62 bp, 50 bp, and 58 bp). Sequence identity and a phylogenetic comparison of the deduced protein (736 aa) with other CPRs in the database suggested that P. chrysosporium CPR is the largest CPR known and is more closely related to animal (36-38%) and yeast (37 38%) CPRs than to plant CPRs (33-35%). The availability of this gene will facilitate further studies on understanding the potent xenobiotic mono oxygenation systems in this model white rot fungus. PMID- 10672449 TI - The cost-effectiveness of different management strategies for type I diabetes: a Swiss perspective. AB - AIMS/HYPOTHESIS: A computer model was developed to determine the health outcomes and economic consequences of different combinations of diabetes interventions in newly diagnosed patients with Type I (insulin-dependent) diabetes in Switzerland. METHODS: We modelled seven complications of diabetes: hypoglycaemia, ketoacidosis, acute myocardial infarction, stroke, lower extremity amputation, nephropathy, and retinopathy. Transition probabilities and costs were taken from published literature. The Swiss health insurance payer perspective was taken. Various combinations of diabetes management strategies, including intensive or conventional insulin therapy and screening and treatment strategies for renal and eye disease were defined. Life expectancy, cumulative incidences of complications, and mean expected total lifetime costs per patient were calculated under six different management strategies. Incremental cost-effectiveness ratios were calculated in terms of costs per life-year gained compared with conventional insulin therapy alone. RESULTS: The addition of screening for microalbuminuria and retinopathy followed by appropriate treatment, if detected, were cost saving, with reduction in cumulative incidence of end stage renal disease and blindness respectively, and, in the case of microalbulminuria screening and treatment, an improvement in life expectancy. Intensive therapy improved life expectancy but increased total lifetime costs. CONCLUSION/INTERPRETATION: Optimal management of Type I diabetic patients, including secondary and tertiary prevention, leads to reduced complications and improved life expectancy, with the increased costs of prevention offset to varying degrees by cost savings due to complications avoided. PMID- 10672450 TI - Effects of diabetic cardiomyopathy on regional electrophysiologic characteristics of rat ventricle. AB - AIMS/HYPOTHESIS: To identify the possible causes of the lengthening of the action potential duration described in patients affected by diabetes mellitus. METHODS: We studied the effects of streptozotocin-induced diabetes on the current density of the repolarising potassium currents It(o), IK, Iss and IK1 in enzymatically isolated myocytes from three different regions of rat heart: total right ventricle, subepicardium at the apex of the left ventricle and subendocardium at the base of the left ventricle. RESULTS: No changes in IK1 were found due to diabetes, but there was a uniform decrease in It(o) (50%) and Iss (40%) current densities in the three regions. In contrast, IK diminished unevenly, with the greatest decrease in the subendocardium at the base of the left ventricle (48%), followed by the subepicardium at the apex of the left ventricle (32%) and right ventricle (10%). CONCLUSION/INTERPRETATION: These findings suggest the existence of regional differences in ion channel expression associated with diabetes. The decrease of these repolarising currents could account for the lengthening of action potential and the consequent change in the Q-T interval of the ECG observed in diabetic rats. PMID- 10672451 TI - Functional and structural abnormalities in the nerves of type I diabetic baboons: aminoguanidine treatment does not improve nerve function. AB - AIMS/HYPOTHESIS: To improve understanding of the pathophysiology of diabetic neuropathy and to establish a primate model for experimental studies, we examined nerve changes in baboons with Type I (insulin-dependent) diabetes mellitus. We also examined the effect of aminoguanidine (an inhibitor of the formation of advanced glycation end products) on nerve function. METHODS: Male baboons (Papio hamadryas) were assigned to four groups; control, diabetic, control and diabetic treated with aminoguanidine. Diabetes was induced with streptozotocin (60 mg/kg, intravenous). Insulin and aminoguanidine (10 mg/kg) were injected subcutaneously daily. Motor and sensory nerve conduction velocity was measured using standard techniques. Autonomic function was examined by measuring heart rate response to positional change. Sural nerve morphometry was analysed in the diabetic group (mean duration 5.5 years) along with their age-matched controls. RESULTS: The diabetic groups were smaller in size with a mean HbA1c of 8.9 +/- 1.2%. The nerve conduction velocity and heart rate response was reduced in the diabetic groups. Morphometric analysis of the diabetic sural nerve showed smaller axon diameter (2.99 +/- 0.06 microns vs 3.29 +/- 0.06 microns; p < 0.01) accompanied by thinner myelin (1.02 +/- 0.02 microns vs 1.15 +/- 0.02 microns, p < 0.01) with no change in the axon density. Treatment with aminoguanidine for 3 years had no effect on glycaemic control and did not restore conduction velocity or autonomic dysfunction in the diabetic animals, contrary to the studies in rats. CONCLUSIONS/INTERPRETATION: These results show that the primate is a good model to study diabetic neuropathy and suggest that the accumulation of advanced glycation end products are not an early mechanism of nerve damage in this disorder. PMID- 10672452 TI - Excessive fat accumulation is associated with the TNF alpha-308 G/A promoter polymorphism in women but not in men. AB - AIMS/HYPOTHESIS: Tumour necrosis factor alpha (TNF alpha) is a candidate gene for the development of obesity, which in turn is a major risk factor for diabetes mellitus. The aim of our study was to investigate whether a previously known NcoI sensitive polymorphism (-308 G/A) in the promoter region of the TNF alpha gene was related to body weight. METHODS: Genotyping was done in 239 male and 342 female non-diabetic subjects with a marked variation in body mass index (BMI). RESULTS: We found three genotypes; AA (n = 13), AG (n = 158) and GG (n = 410). When the material was divided according to sex, allele specific phenotypic differences were confined to women. The female subjects carrying the AA genotype were markedly more obese than both the AG and GG carriers (mean BMI = 41.4 vs 32.3 and 31.7 kg/m2, respectively, p = 0.02). The body fat content of female AA carriers was increased by one-third compared with AG/GG carriers (p = 0.02). We found no differences between genotypes with respect to waist-to-hip ratio, blood pressure or metabolic variables. Among obese female subjects (BMI > 27 kg/m2), we also found that the BMI and body fat content of AA carriers (n = 7) were also higher than for AG/GG carriers. CONCLUSION/INTERPRETATION: The AA-variant at position -308 in the promoter region of the TNF alpha gene could be an important genetic factor behind excessive fat accumulation in women. PMID- 10672453 TI - Cloning of cDNA and the gene encoding human hepatocyte nuclear factor (HNF)-3 beta and mutation screening in Japanese subjects with maturity-onset diabetes of the young. AB - AIMS/HYPOTHESIS: Molecular defects of the genes for transcription factors, hepatocyte nuclear factor (HNF)-4 alpha, HNF-1 alpha, HNF-1 beta and insulin promoter factor-1 cause maturity-onset diabetes of the young (MODY1, 3, 5, and 4, respectively). This suggests the HNF-related transcription cascade is important in insulin secretion which is induced by glucose. These genes and the gene encoding glycolytic enzyme glucokinase (MODY2) are, however, responsible for only 15-20% of cases of MODY in the Japanese. Searching for a novel form of MODY in this population, we cloned a new candidate gene encoding human HNF-3 beta, a winged helix transcription factor, which also belongs to the same HNF transcription cascade. METHODS: The cDNA clone for human HNF-3 beta was isolated from a liver cDNA library. The gene was also cloned from a genomic library and its organization and chromosomal localization were determined. We screened 68 Japanese subjects with MODY/early-onset diabetes for mutations in this gene. RESULTS: Human HNF-3 beta is composed of 457 amino acids. The human gene, which was mapped to the segment 30 cR from SHGC-37039 on chromosome 20p by radiation hybrid mapping, spans approximately 4.5 kb and consists of three exons. Direct sequencing of the exons and flanking regions identified one missense mutation A328 V and seven polymorphisms, although the functional significance of the mutation in the pathogenesis of diabetes is not known. CONCLUSION/INTERPRETATION: The characterization of the structure of the HNF-3 beta gene and its mapping in the framework of markers will be helpful in genetic studies of the various forms of diabetes mellitus. PMID- 10672454 TI - Are randomized controlled trials sufficient evidence to guide clinical practice in type II (non-insulin-dependent) diabetes mellitus? AB - Randomized controlled trials (RCTs) are often considered the standard for defining the practice of evidence-based medicine. Taken alone, they are, however, often insufficient to guide clinical care. Randomized controlled trials are clearly the best method to determine whether interventions are efficacious. They have, however, numerous limitations which make them difficult to carry out or limit applicability to routine clinical practice. Although observational studies also have inherent limitations, they provide data which can help to further explain the results of randomized controlled trials. The use of observational studies to frame randomized trials can allow better application of randomized controlled trial results to individual patients and can thus help to optimize delivery of care, inform clinical practice and determine the need for further such trials. PMID- 10672455 TI - Hepatocyte nuclear factor-1 beta (MODY5) gene mutations in Scandinavian families with early-onset diabetes or kidney disease or both. PMID- 10672456 TI - Accuracy of fasting glucose to diagnose diabetes in Brazilian subjects. PMID- 10672457 TI - Comments on the relation among GAD-65, IA-2 antibodies and body adiposity. PMID- 10672458 TI - 125 years of enantiomers--back to the roots Jacobus Henricus van't Hoff 1852 1911. PMID- 10672459 TI - Developing a chiral toolbox for asymmetric catalytic reactions. AB - During the last several decades, chemists have made major progress in discovering man-made catalysts to perform challenging asymmetric transformations. The research in our group addresses fundamental and practical problems in this field by developing a diverse set of chiral ligands that combine with transition metals to form highly enantioselective catalysts. Families of tridentate ligands have been developed for the enantioselective hydrogenation of unfunctionalized substrates. In addition, we have developed several new bidentate phosphine ligands for asymmetric catalysis. The common feature of these ligands are that they contain rigid aromatic backbones or ring structures which restrict conformational flexibility of the ligands. Several asymmetric reactions have been studied: asymmetric hydrogenation of functionalized substrates such as N acylaminoacrylic acids, enamides and enol acetates; asymmetric hydrogenation of simple ketones, and imines; and asymmetric carbon-carbon bond forming reactions. In addition, we have designed and synthesized several novel chiral monophosphines for asymmetric catalytic reactions. Transition metal complexes with these monophosphines have also been developed for asymmetric transformations. PMID- 10672460 TI - Stereoselective conjugate additions to enoates. PMID- 10672461 TI - Chemical process evolution of efavirenz, a potent non-nucleosidal HIV reverse transcriptase inhibitor. PMID- 10672462 TI - The origins of heterogeneous catalysis by platinum: Johann Wolfgang Dobereiner's contributions. AB - A number of examples of what Jons Jacob Berzelius was later (1835) to call catalysis, have been known from antiquity. One of the most prominent of these, dating from the early eighteenth century, was the action of platinum black on hydrogen observed in 1823 by Johann Wolfgang Dobereiner (1780-1849), Professor of Chemistry and Technology at the Universitat Jena (1810-1849), who is regarded as the discoverer of platinum catalysis. His startling discovery led to his invention of the widely used pneumatic gas lighter (Dobereinersches Feuerzeug) and was cited by Berzelius in his formulation of the concept of catalysis. PMID- 10672463 TI - Control of enantioselectivity in catalytic metal carbene reactions. AB - Chiral dirhodium(II) complexes constructed from 2-oxopyrrolidine, 2 oxazolidinone, N-acyl-2-imidazolidinone, or 2-azetidinone ligands are exceptional catalysts for enantioselective metal carbene transformations which provide lactones and lactams via cyclopropanation, cyclopropenation, C-H insertion, and ylide reactions with enantiomeric excesses greater than 90% and, usually, in very high yields. These chiral catalysts present high diastereo-, regio- and chemoselectivities, turnover numbers up to 1000, as well as recoverability and reuse. PMID- 10672464 TI - Mapping of c-Fos in the trigeminal sensory nucleus following high- and low intensity afferent stimulation in the rat. AB - Although previous studies have suggested that nociceptive afferents from intra oral and facial structures are organized differently in the trigeminal sensory nucleus (TSN), more detailed data are needed. The present study aimed to fill this gap, by examining the changes in the expression of c-Fos within the rat TSN following high- and low-intensity electrical stimulation applied to the Gasserian ganglion (GG). A low-intensity stimulus (0.1 mA) induced c-Fos in many neurons in the dorsomedial subdivision (Vodm) of the oral subnucleus (Vo; mean +/- SEM in a certain segment = 163.0 +/- 42.7), in the medial part of the dorsomedial subdivision (Vidm) of the interpolar subnucleus (Vi; 120.5 +/- 40.1), in the medial corner of the magnocellular zone (VcIII/IV; 47.5 +/- 10.5), and in the superficial layers (VcI/II; 1330.0 +/- 65.6) along the entire length of the dorsomedial-ventrolateral axis of the caudal subnucleus (Vc). A modest number of Fos-positive neurons were induced in the dorsal principal subnucleus (Vp; 10.0 +/ 4.9) and in the lateral VcIII/IV (11.5 +/- 1.6). A high-intensity stimulus (1.0 mA) significantly increased the number of Fos-positive neurons in each subdivision compared with the low-intensity stimulus (Vp 32.3 +/- 10.8; Vodm 270.3 +/- 75.3; Vidm 189.3 +/- 38.5; medial VcIII/IV 77.5 +/- 18.2; lateral VcIII/IV 24.8 +/- 9.3; VcI/II, 2155.8 +/- 470.2). At both low- and high-intensity stimulation, the fields where Fos-positive neurons appeared are restricted to the dorsal or dorsomedial subdivisions of the rostral subnuclei, Vp, Vo and Vi, where the main projectional fields of primary afferents from the intraoral structures are found, while Fos-positive neurons were distributed in the entire VcI/II, along the dorsomedial-ventrolateral axis of Vc, where the main projectional fields of primary afferents from the facial skin are found. The threshold to induce c-Fos is, however, different according to the fields. These results suggest that nociceptive processing in the intra-oral region is mediated through the entire length of the rostro-caudal axis of TSN, but is mediated primarily through VcI/II in the facial region. PMID- 10672465 TI - Effects of stimulus size and eccentricity on horizontal and vertical vergence. AB - We measured the gain and phase of horizontal and vertical vergences of five subjects as a function of stimulus area and position. Vergence eye movements were recorded by the scleral search coil method as subjects observed dichoptic displays oscillating in antiphase either from side to side or up and down with a peak-to-peak magnitude of 0.5 degree at either 0.1 Hz or 1.0 Hz. The stimulus was a central textured disc with diameter ranging from 0.75 degree to 65 degrees, or a peripheral annulus with outer diameter 65 degrees and inner diameter ranging from 5 degrees to 45 degrees. The remaining field was black. For horizontal vergence at both stimulus frequencies, gain and the phase lag were about the same for a 0.75 degree stimulus as for a 65 degrees central stimulus. For vertical vergence, mean gain increased and mean phase lag decreased with increasing diameter of the central stimulus up to approximately 20 degrees. Thus, the stimulus integration area is much smaller for horizontal vergence than for vertical vergence. The integration area for vertical vergence is similar to that for cyclovergence, as revealed in a previous study. For both types of vergence, response gains were higher and phase lags smaller at 0.1 Hz than at 1.0 Hz. Also, gain decreased and phase lag increased with increasing occlusion of the central region of the stimulus. Vergence gain was significantly higher for a 45 degrees central disc than for a peripheral annulus with the same area. Thus, the central retina has more power to evoke horizontal or vertical vergence than the same area in the periphery. We compare the results with similar data for cyclovergence and discuss their ecological implications. PMID- 10672466 TI - Instructing subjects to make a voluntary response reveals the presence of two components to the audio-vocal reflex. AB - Previous findings have shown that subjects respond to an alteration, or shift, of auditory feedback pitch with a change in voice fundamental frequency (F0). When pitch shifts exceeding 500 ms in duration were presented, subjects' averaged responses appeared to consist of both an early and a late component. The latency of the second response was long enough to be produced voluntarily. To test the hypothesis that there are two responses to pitch-shift stimuli and to clarify the role of intention, subjects were instructed to change their voice F0 in the opposite direction of the pitch-shift stimulus, in the same direction, or not to respond at all. In a second group, subjects were tested under the above conditions as well as under instructions to raise voice F0 or to lower F0 as rapidly as possible upon hearing a pitch shift. Results showed that, when given instructions to produce a voluntary response, subjects made both an early vocal response (VR1) and a later vocal response (VR2). The second response, VR2, was almost always made in the instructed direction, whereas VR1 was often made incorrectly. The latency of VR1 was reduced under instructions to respond to feedback pitch shifts by changing voice F0 in the opposite direction, compared with that when told to ignore the pitch shifts. Latency and amplitude measures of VR2 differed under the various experimental conditions. These results demonstrate that there are two responses to pitch-shift stimuli. The first is relatively automatic but may be modulated by instructions to the participant. The second response is probably a voluntary one. PMID- 10672467 TI - Effects of systemic 3-nitropropionic acid-induced lesions of the dorsal striatum on cannabinoid and mu-opioid receptor binding in the basal ganglia. AB - Systemic administration of 3-nitropropionic acid (3NPA) in experimental animals produces bilateral striatal lesions similar to those seen in Huntington's disease (HD) caudate and putamen. 3H[-CP55,940 binding to cannabinoid receptors in human basal ganglia nuclei has been shown to be highly susceptible to the earliest pathological changes in the HD brain. In this study, to assess further the suitability of 3NPA-induced striatal lesions as a model for HD neuropathology, we examined the effects of striatal lesions induced by the systemic administration of 3NPA on the binding of 3H[-CP55,940 to pre- and postsynaptic cannabinoid receptors in striatum, globus pallidus, entopeduncular nucleus and substantia nigra pars reticulata and also the effect of 3NPA-induced striatal lesions on the binding of 3H[-DAMGO to mu-opioid receptors in striatal striosomes. Systemic administration of 3NPA induced bilateral and symmetrical lesions in dorsolateral striatum. Within the lesion core, 3H[-CP55,940 and 3H[-DAMGO binding density was reduced to background levels. Beyond the immediate borders of the central core of the 3NPA-induced lesion, striatal binding density was not significantly different from that measured in unlesioned rats. 3H[-CP55,940 binding in globus pallidus, entopeduncular nucleus and substantia nigra in 3NPA-lesioned rats was significantly reduced compared to controls, and the individual decreases were similar for each site. However, these reductions were statistically marginal. These data suggest that, while producing striatal lesions which bear some similarity to those seen in HD, the consequences of 3NPA for striatopallidal and striatonigral efferent projections do not reflect the reported neurodegenerative changes seen in the HD brain. PMID- 10672468 TI - Responses of vestibular nucleus neurons to tilt following chronic bilateral removal of vestibular inputs. AB - Recordings were made from the vestibular nuclei of decerebrate cats that had undergone a combined bilateral labyrinthectomy and vestibular neurectomy 49-103 days previously and allowed to recover. Responses of neurons were recorded to tilts in multiple vertical planes at frequencies ranging from 0.05 to 1 Hz and amplitudes up to 15 degrees. Many spontaneously active neurons were present in the vestibular nuclei; the mean firing rate of these cells was 43 +/- 5 (SEM) spikes/s. The spontaneous firing of the neurons was irregular: the coefficient of variation was 0.86 +/- 0.14. The firing of 27% of the neurons was modulated by tilt. The plane of tilt that elicited the maximal response was typically within 25 degrees of pitch. The response gain was approximately 1 spikes/s/degree across stimulus frequencies. The response phase was near stimulus position at low frequencies, and lagged position slightly at higher frequencies (average of 35 +/ 9 degrees at 0.5 Hz). The source of the inputs eliciting modulation of vestibular nucleus activity during tilt in animals lacking vestibular inputs is unknown, but could include receptors in the trunk or limbs. These findings show that activation of vestibular nucleus neurons during vertical rotations is not exclusively the result of labyrinthine inputs, and suggest that limb and trunk inputs may play an important role in graviception and modulating vestibular elicited reflexes. PMID- 10672469 TI - Voluntary modification of automatic arm movements evoked by motion of a visual target. AB - We have investigated whether the processes underlying the visually evoked, automatic adjustments to a reach are: (1) modifiable by the subject's intention, and (2) available to initiate movement of a stationary arm. Unpredictable movement of a target (80 m/s through 10 cm, left or right in a third of trials) either evoked a mid-flight adjustment of a reaching movement or else acted as a trigger to start an arm movement. Subjects were instructed to respond as rapidly as possible by moving their finger either in the same or in the opposite direction to the target. The target shift evoked an early (125-160 ms) and/or a later (> 160 ms) class of response in the reaching arm. The early response was highly automatic in that it could not be reversed (move opposite) by the subjects' intention. However, the subjects' intention did influence the frequency of occurrence and the size of this early response. The later response was totally modifiable in that it changed direction according to the subjects' intention. Similar classes of response were observed in stationary limbs, but the early, more automatic response was substantially weaker than that elicited during a reach. Two possible mechanisms are proposed to explain these results. The first is a dual-pathway model, which assumes that the two response classes are each generated by separate visuo-motor processes with different properties. The second model assumes both responses are generated by a single visuo-motor mechanism that is under the control of a higher, attentional process. PMID- 10672470 TI - Flexibility of saccade adaptation in the monkey: different gain states for saccades in the same direction. AB - Saccadic accuracy, measured as the ratio of the size of a saccade to the size of the target step that elicits it, i.e., saccade gain, can be altered by jumping the target surreptitiously during the targeting saccade. The gain change produced by this paradigm does not generalize or transfer to saccades of all sizes. Instead, the amount of transfer decreases the more the tested saccade differs in amplitude and direction from that adapted. Here, we tested the limits of this saccade-size specificity by attempting to impose quite different gain states on saccades in the same direction. We altered the saccadic gain by intrasaccadic target jumps of 30% of the initial target step, either forward to produce a gain increase or backward to produce a gain decrease. Three different conditions were studied: (1) saccades to target steps of 20 degrees or 7 degrees were adapted in individual sessions with backward and forward jumps, respectively; (2) saccades to target steps of 20 degrees caused backward target jumps during the same session in which saccades to 7 degrees target steps caused forward steps; (3) the target jumps accompanying 20 and 7 degrees saccades were the same as in (2), but in addition, there were intermediate-sized saccades to 13.5 degrees target steps with no intrasaccadic target jumps. Saccadic gain adaptation was quite flexible. In condition 2, we could simultaneously increase the gain of saccades to 7 degrees target steps while decreasing the gain of saccades to 20 degrees steps in the same direction. Intermediate horizontal saccades to 13.5 degrees target steps experienced gain reductions (average: 6.9%), which were not the sum of gain changes expected from separate 20 degrees gain decreases and 7 degrees gain increases alone, as predicted from condition 1. If adaptation at 20 degrees and 7 degrees occurred while an animal also tracked a non-adapting 13.5 degrees target step (paradigm 3), the gain reduction of saccades to the 13.5 degrees step was reduced considerably (3.4%). Thus, the mechanism that adapts saccade size can support a robust gain increase for saccades of one size while simultaneously supporting a robust gain decrease for saccades only 13 degrees larger. Furthermore, the presence during adaptation of a non-adapted target step with a size intermediate to the two adapting steps reestablishes a nearly normal gain within only 6.5 degrees of a robust gain increase and decrease. These data indicate that saccadic gain adaptation can set very different gain states for saccades with rather similar vectors. PMID- 10672471 TI - Electrophysiological correlates of human intrasaccadic processing. AB - Visual discrimination performance is thought to be suppressed during saccades in order to contribute to space constancy. However, under certain experimental conditions, visual inhibition may not take place, suggesting a more complex underlying mechanism. We tested the discrimination ability of 20 healthy subjects during visually guided horizontal saccades and recorded simultaneously the evoked brain activity from 30 channels over occipital, parietal, and temporal areas. During the execution of saccadic eye movements, visual stimuli were presented for 30 ms. In order to prevent retinal afterimages, stimuli were followed by a visual mask. In a control condition, the same stimuli were presented with stationary eyes. Electro-oculogram (EOG) and electroencephalogram (EEG) signals were recorded continuously together with information about the stimuli and the subject's response. Evoked potentials were computed offline, and component latency, field strength (global field power), and topography were compared between conditions. During saccades, subjects showed only slightly reduced discrimination performance which remained very high above the chance level; thus, there was no evidence for strong saccadic suppression with the supra-threshold stimuli employed. However, the cortical activation patterns exhibited large alteration when a physically identical stimulus was presented during the eye movement: around 130 ms latency, field strength was significantly smaller than when stationary targets were processed, and scalp topography was also different. These effects on evoked field distributions may be attributed to neural interactions of an efference copy signal (linked to the oculomotor command) with the afferent excitation following the visual stimulus. PMID- 10672472 TI - Descending corticofugal neurons in layer 5 of rabbit S1: evidence for potent corticocortical, but not thalamocortical, input. AB - Extracellular recordings were obtained from descending corticofugal neurons of layer 5 (CF-5 neurons) in primary somatosensory cortex (S1) of awake rabbits. These cells were identified by antidromic activation via stimulation sites in ventrobasal (VB) thalamus. Recordings were also obtained from putative GABA-ergic interneurons (suspected inhibitory interneurons, SINs) located in the same microelectrode penetrations, and in close proximity (+/- 300 microns) to the CF-5 neurons. In some experiments, the above populations were recorded simultaneously with neurons in the topographically aligned VB thalamic barreloid. Each of several experimental strategies failed to reveal evidence of monosynaptic thalamic input to CF-5 neurons, but revealed a clear monosynaptic input to neighboring SINs: (1) whereas CF-5 neurons responded at very long synaptic latencies to intense electrical stimulation of VB thalamus, neighboring SINs responded at short latencies; (2) whereas cross-correlations between CF-5 neurons and topographically aligned VB neurons failed to show significant peaks indicative of monosynaptic VB input, neighboring SINs did show such peaks; and (3) whereas CF-5 neurons were unresponsive to microstimulation of topographically aligned VB thalamic barreloids, neighboring SINs were very responsive to such stimulation. Both CF-5 neurons and neighboring SINs responded to electrical stimulation of the corpus callosum with a robust, short-latency synaptic response. This finding demonstrates that CF-5 neurons are capable of vigorous, short-latency responses to excitatory synaptic input. These data suggest considerable specificity in the thalamocortical connectivity of subpopulations within layer 5, and support the notion that CF-5 neurons are dominated by corticocortical rather than thalamocortical input. PMID- 10672473 TI - Oscillations in the premotor cortex: single-unit activity from awake, behaving monkeys. AB - We examined single-unit activity in the dorsal premotor cortex for evidence of fast neuronal oscillations. Four rhesus monkeys performed a task in which visuospatial instruction stimuli indicated the direction of forelimb movement to be executed on each trial. After an instructed delay period of 1.5-3 s, movements to either the right or left of a central origin were triggered by a second visuospatial stimulus. From a database of 579 single units, 78 units (13%) contained periodic peaks in their autocorrelation histograms (ACHs), with oscillation frequencies typically 20-30 Hz (mean 27 Hz). An additional 26 units (5%) had oscillatory features that were identified in joint interspike-interval (ISI) plots. Three observations, taken together, suggest entrainment by rhythmic drive extrinsic to these neurons: shuffling ISIs attenuated ACH peaks, indicating a dependency on serial-order effects; oscillation frequency did not change during either increases or decreases in firing rate; and joint ISI plots contained features consistent with a rhythmicity interrupted by intervening discharges. In some cells, oscillations occurred for only one of the two directions of movement. During the delay period, such directional selectivity was observed in 37 units (60% of delay-period oscillators). For at least 17 of these units, we could exclude the possibility that oscillatory directional selectivity resulted from the difficulty in detecting oscillations due to low discharge rates (for one of the two movement directions). Directional selectivity in fast oscillations shows that they can reflect specific aspects of an intended action. PMID- 10672474 TI - Transfer of tritiated water, tyrosine, and propanol from the nasal cavity to cranial arterial blood in rats. AB - Respiration cools the nasal vein blood. This may, via counter current transfer in the cavernous sinus/carotid artery complex, cool the brain arterial blood and, therefore, decrease the brain temperature during heat stress. We investigated the possible local transfer of substances from the nasal cavities to the brain via a similar transfer between nasal venous blood and brain arterial blood. Tritiated water (TW), 3H-tyrosine (T) and 14C-propanol (P) were infused into the nasal cavity of anesthetized rats that had two catheters inserted into the same, occluded carotid artery, one pointing towards the head, the other towards the heart. Continuous, parallel blood samples were collected throughout 30-s periods for 10 min, and the radioactivity measured in the separated plasma. After nasal application of TW, the radioactivity increased in the head arterial plasma samples compared with the parallel heart samples. When recirculation to the general circulation was diminished, a larger and significant difference was found for all three substances during the last 5 min of the collection period. No difference between the parallel samples was observed after intravenous administration of the three substances. Absorption of all three substances from the nasal cavity was rapid and high. The results indicate that local transfer takes place between the venous and arterial blood in the head, probably in the area of the cavernous sinus-carotid artery complex, which in rat and man lacks a Rete Mirabile. The local transfer raises the possibility of treating brain diseases in man locally through nasal application of drugs. PMID- 10672475 TI - Striate cortex in humans demonstrates the relationship between activation and variations in visual form. AB - Electrophysiologic and functional imaging studies have shown that the visual cortex produces differential responses to the presence or absence of structure within visual textures. To further define and characterize regions involved in the analysis of form, functional magnetic resonance imaging (fMRI) was used to detect changes in activation during the viewing of four levels of isodipole textures. The texture levels systematically differed in the density of visual features such as extended contours and blocks of solid color present within the images. A linear relationship between activation level and density of structure was observed in the striate cortex of human subjects. This finding suggests that a special subpopulation of striate cortical neurons participates in the ability to extract and process structural continuity within visual stimuli. PMID- 10672476 TI - Activity in deep intermediate layer collicular neurons during interrupted saccades. AB - The activity of neurons located in the deep intermediate and adjacent deep layers (hereafter called just deep intermediate layer neurons) of the superior colliculus (SC) in monkeys was recorded during saccades interrupted by electrical stimulation of the brainstem omnipause neuron (OPN) region. The goal of the experiment was to determine if these neurons maintained their discharge during the saccadic interruption, and thus, could potentially provide a memory trace for the intended movement which ends accurately on target in spite of the perturbation. The collicular neurons recorded in the present study were located in the rostral three-fifths of the colliculus. Most of these cells tended to show considerable presaccadic activity during a delayed saccade paradigm, and, therefore, probably overlap with the population of SC cells called buildup neurons or prelude bursters in previous studies. The effect of electrical stimulation in the OPN region (which interrupted ongoing saccades) on the discharge of these neurons was measured by computing the percentage reduction in a cell's activity compared to that present during non-interrupted saccades. During saccade interruption about 70% of deep intermediate layer neurons experienced a major reduction (30% or greater) in their activity, but discharge recovered quickly after the termination of the stimulation as the eyes resumed their movement to finish the saccade on the target. Therefore, the pattern of activity recorded in most of the deep intermediate layer neurons during interrupted saccades qualitatively resembled that previously reported for the saccade-related burst neurons which tend to be located more dorsally in the intermediate layer. In contrast, some of our cells (30%) showed little or no perturbation in their activity caused by the saccade interrupting stimulation. Because all the more dorsally located burst neurons and the majority of our deep intermediate layer neurons show a total or major suppression in their discharge during interrupted saccades, it seems unlikely that the colliculus by itself could maintain an accurate memory of the desired saccadic goal or the remaining dynamic motor error required to account for the accuracy of the resumed movement which occurs following the interruption. However, it remains possible that the smaller proportion of our neurons whose activity was not perturbed during interrupted movements could play a role in the mechanisms underlying saccade accuracy in the interrupted saccade paradigm. Interrupted saccades have longer durations than normal saccades to the same target. Therefore, we investigated whether the discharge of our deeper collicular cells was also necessarily prolonged during interrupted saccades, and, if so, how the prolongation compared to the prolongation of the saccade. Sixty percent of our sample neurons showed a prolongation in discharge that was approximately the same as the prolongation in saccade duration (difference < 15 ms in magnitude). The, observation that temporal discharge in our neurons was perturbed to roughly match saccadic temporal perturbation suggests that dynamic feedback about ongoing saccadic motion is provided to the colliculus, but does not necessarily imply that this structure is the site responsible for the computation of dynamic motor error. PMID- 10672477 TI - Functional reorganisation of the corticomotor projection to the hand in skilled racquet players. AB - While it is known that relatively rapid changes in functional representation may occur in the human sensorimotor cortex in short-term motor-learning studies, there have been few studies of changes in organisation of the corticomotor system associated with the long-term acquisition of motor skills. In the present study, we have used transcranial magnetic stimulation (TMS) to investigate the corticomotor projection to the hand in a group of elite racquet players, who have developed and maintained a high level of skill over a period of many years, and have compared the findings with those in a group of social players and a group of non-playing control subjects. Increased motor-evoked-potential (MEP) amplitudes and shifts in the cortical motor maps for the playing hand were found in all of the elite players and cortical motor thresholds were reduced in some players, whereas in the social players all parameters were within the normal range. The findings in the elite players are interpreted as being indications of a process of functional reorganisation with the motor cortex or corticomotor pathway that are associated with the acquisition and retention of complex motor skills. PMID- 10672478 TI - Cooling of the brain through oxygen flushing of the nasal cavities in intubated rats: an alternative model for treatment of brain injury. AB - Local cooling of the brain by respiration has been found in several animal species with a rete mirabile in the carotid artery/cavernous sinus complex. The present experiment was made to investigate whether a similar cooling could be found in the rat, which does not have a rete. Eleven rats were anesthetized and intubated. Three thermoprobes were inserted into the brain (two probes) and rectum, and the temperatures measured continuously. The nasal cavities were flushed with oxygen (250-1000 ml/min) during 15-min periods, interrupted by 15 min control periods. The mean brain temperature decreased by 0.43 +/- 0.03 degree C (n = 86, P < 0.005) with individual values up to 1.11 degrees C during the flushing periods. The decrease was oxygen-flow dependent, but not correlated to the rectal temperature. It is concluded that even an animal species without a rete mirabile is able to decrease the brain temperature through nasal cooling. The cooling was probably connected to the blood flow. If the results can be extrapolated to man (no rete mirabile), brain temperature can be decreased by nasal flushing with air or oxygen in intubated patients with hyperthermia. We also suggest that this simple treatment will reduce the infarct volume after head injury, trauma, or brain ischemia. PMID- 10672479 TI - Intentional on-line adaptation of stride length in human walking. AB - The intentional control of stride length is a fundamental basis for the adaptation of the stride to environmental constraints (obstacle avoidance, for example). Controlling the propulsive forces during the stance and/or controlling the pendular movement of the oscillating leg constitute the two potential and non exclusive mechanisms underlying intentional stride length modulation. The present experiment was conducted in order to determine if these two mechanisms contribute to voluntary length modulation and, if so, how they cooperate according to whether the subject has to lengthen or shorten a stride and how these mechanisms are implemented at the neuromuscular level. Subjects had to produce a temporarily modulated stride of the same length, but originating from two different initial steady-states: one from shorter stride length and one from longer stride length. We found that the shortening was essentially realized by a swing-duration decrease (an increased activity in the hip extensor--biceps femoris--during the swing of the ipsilaterally shortened stride stopped the pendular leg movement earlier). The lengthening was realized by two mechanisms: (1) an increase in the propulsive forces (via an increased activity of the ankle extensor muscles- soleus--and the hip extensors--biceps femoris--from the stance of the ipsilaterally modulated stride, which was prolonged during the following stance of the contralateral leg), and (2) an increase in swing duration on the ipsilateral leg (an increased activity in hip and ankle flexors--rectus femoris and tibialis anterior--maintained the ipsilateral leg in flexion during the lengthened swing so that the foot landed later). In this experiment, the subjects were faced with a spatial constraint of the same magnitude in the direction of stride lengthening and stride shortening. However, under these conditions, subjects used a different balance between swing control (that directly modifies the foot trajectory without affecting the trajectory of the head-arm-trunk system) and/or the control of propulsive forces (that indirectly influences foot trajectory by modifying the trajectory of the head-arm-trunk system). In the first case, this concerns a voluntary control of gesture produced by the legs and usually implicated in the locomotor pointing; in the second case, this concerns a voluntary control of propulsive forces. PMID- 10672480 TI - Estimating the components of the gap effect. AB - The gap effect refers to the finding that saccadic latencies are typically reduced when a fixation point is removed prior to the appearance of a peripheral target. This reduction in saccadic reaction time (SacRT) is thought to be due to a general warning effect and an oculomotor specific fixation offset that occur when the fixation point is removed. In order to estimate the contribution of each of these effects to the overall gap effect, this paper introduces a new manipulation, the partial-gap trial, where the fixation point undergoes a change in size prior to the presentation of the target. The partial-gap trial is presumed to provide the visual warning effect of the fixation offset (i.e. similar to that in a gap trial) but does not provide the fixation offset effect (FOE). When the fixation point was abruptly reduced in size before the presentation of the target, the estimated decrease in SacRT due to the visual warning effect was 5-7% and did not differ in the presence or absence of an auditory warning signal. It was found that auditory warning effect and the FOE interacted in reducing SacRTs. Additionally, when the fixation point was abruptly increased in size before the presentation of the target, SacRTs were slower than when the fixation point did not change in size and remained present for the entire trial (i.e. an overlap trial). We conclude that this new partial-gap paradigm is a useful method for researchers wishing to separately examine FOE and visual warning effects. PMID- 10672481 TI - Inhibition of return in saccadic eye movements. AB - Inhibition of return (IOR) is a phenomenon in which responses generated to targets at previously attended locations are delayed. It has been suggested that IOR affords a mechanism for optimizing the inspection of novel locations and that it is generated by oculomotor reflexes mediated by the superior colliculus. In this investigation, we measured the effects of IOR on the metrics of saccadic eye movements made to novel and previously attended locations. Saccades made to cued target locations, as well as to other targets within the same hemifield, had longer latencies than saccades made towards the novel, uncued hemifield. We further found that the amplitudes of saccades towards the cued hemifield were more hypometric, but only when the amplitude could not be pre-programmed. These results provide evidence that IOR influences spatial, as well as temporal, parameters of saccadic eye movements and suggest that the exogenous orienting of attention, in addition to influencing target detection, also influences oculomotor programming. PMID- 10672482 TI - Visual and haptic feedback in the control of force. AB - The ability to control index finger and elbow flexion forces was measured while subjects used either haptic feedback or both haptic and visual feedback to control the forces exerted. Over a 120-s time period subjects were able to control the finger forces ranging from 2 to 6 N to within 1 N using only haptic feedback, and elbow flexion forces to within 4.5 N over a force range of 10-30 N. At the same force amplitude there was no significant difference between the two muscle groups in the precision or accuracy with which the force could be controlled, suggesting that there is not a proximal to distal gradient in force control as has been found for the control of limb movement and position. PMID- 10672483 TI - The Judd illusion: evidence for two visual streams or two experimental conditions? AB - In the Judd illusion, observers inaccurately bisect the shaft located between two arrowheads pointing in the same direction. The magnitude of error is greater when verbal judgements are compared to action based responses (reaching out and grasping the centre of the bar). This difference has been attributed to the presence of two visual streams within cortical processing. In contrast, we provide evidence that the improved accuracy in the reaching condition may be due to occlusion of the illusory background during the transport phase of the movement. We suggest that caution is required when interpreting performance differences between two conditions that are not strictly equivalent. PMID- 10672484 TI - [Different clinical courses in borreliosis are caused by different pathogens]. PMID- 10672485 TI - Occupational and environmental medicine in Sweden. AB - Great changes have taken place in the fields of occupational and environmental medicine in Sweden during the past decade. Unemployment, work stress, and indoor climate problems have become increasingly common. Chemical exposures in industry and serious work accidents have continued to decrease. State subsidies to occupational health services have been withdrawn and the legislation concerning, for example, occupational diseases has been changed in order to decrease state expenditure. Research has focused on, for instance, ergonomic and psycho-social factors, electromagnetic fields and exposures causing allergy. There is a growing awareness of the need for more emphasis on prevention, which should act in favour of both hospital-based occupational and environmental medicine and the occupational health services sector. PMID- 10672486 TI - Environmental epidemiology--strengths and weaknesses. PMID- 10672487 TI - Possible effects of environmental cadmium exposure on kidney function in the Japanese general population. AB - OBJECTIVES: To examine whether the current level of environmental exposure to cadmium (Cd) is associated with kidney dysfunction among general populations in Japan. METHODS: A nationwide survey was conducted in Japan from 1991 to 1997 at 30 survey sites (with no known environmental heavy metal pollution), by the collection of 24-h food-duplicate samples, peripheral blood specimens and morning spot urine samples. In practice, 607 non-smoking adult women provided these samples. After being wet-ashed, the samples were analyzed for Cd in food duplicates (Cd-F), in blood (Cd-B) and urine (Cd-U) by inductively-coupled plasma mass spectrometry (ICP-MS). Urine samples were also analyzed for alpha 1 microglobulin (alpha 1-MG), beta 2-microglobulin (beta 2-MG) and retinol-binding protein (RBP), creatinine (cr) and specific gravity. Possible tubular dysfunction in association with Cd exposure was examined by simple, multiple and logistic regression analyses, and comparison among three different Cd-dose groups. To minimize the confounding effects of aging, 367 women from 41 to 60 years old were selected and subjected to the same statistical analyses. RESULTS: The analysis of a whole population of 607 women showed that alpha 1-MG and possibly beta 2-MG increased as a function of Cd-F, Cd-B and Cd-U. When the analysis was repeated with the selected population of 367 women aged 41-60, the Cd dose-dependent changes in alpha 1-MG and beta 2-MG became less evident. The distribution of the selected population with alpha 1-MG above two low cut-off values of > 4.9 and > 8.4 mg/g cr or with beta 2-MG above the lowest cut-off value of > 400 micrograms/g cr, was biased toward the group with higher Cd-Ucr, but such bias was not significant for both alpha 1-MG and beta 2-MG when higher cut-off values were employed. No bias was detected with RBP. Logistic regression analysis with alpha 1-MG, beta 2-MG and RBP (with cut-off values given above) in combination with age, Cd-F, Cd-B and Cd-Ucr gave essentially the same results. CONCLUSIONS: The evidence for kidney dysfunction was of borderline significance in the present study population for which geometric mean Cd-F, Cd-B and Cd-U were 24.7 micrograms/day, 1.76 micrograms/l, and 3.94 micrograms/g cr, respectively. The findings might suggest at the same time that the safety margin is small for the Japanese general population regarding environmental Cd exposure. PMID- 10672488 TI - Cadmium exposure of women in general populations in Japan during 1991-1997 compared with 1977-1981. AB - OBJECTIVES: The Japanese people are known to have high environmental exposure to cadmium (Cd). The present survey was initiated to elucidate possible changes in the intensity of Cd exposure to the population by comparison of the present exposure level with the situation some 15 years ago. METHODS: During 1991-1997, 24-h food-duplicate samples, peripheral blood specimens and morning spot urine samples were collected from 588 non smoking women from 27 survey sites in six regions, where food-duplicate and blood samples had also been obtained during 1977-1981 from 399 women. The samples were wet-ashed (after homogenization in the case of food-duplicates), and Cd in the wet-ashed samples was analyzed by inductively-coupled plasma mass spectrometry for Cd intake via foods (Cd-F), Cd concentration in blood (Cd-B) and Cd concentration in urine (Cd-U). The Cd-F and Cd-B were compared with the Cd-F and Cd-B obtained at the same sites in the 1977 1981 survey. RESULTS: The exposure levels during 1991-1997 were such that Cd-F, Cd-B and Cd-Ucr (Cd-U after correction for creatinine concentration) were 25.5 micrograms/day, 1.90 micrograms/l and 4.39 micrograms/g creatinine. Comparison with the 1977-1981 survey results (i.e., 37.5 micrograms/day for Cd-F and 3.47 micrograms/l for Cd-B) showed that there were significant reductions (by 32 and 45%) in both parameters respectively during the last 15 years. The dietary route was an almost exclusive (i.e., 99% of the sum of dietary and respiratory uptake) route of Cd uptake, of which Cd in rice (11.7 micrograms/day) contributed about 40% of the total dietary intake. When compared among survey sites, inter-site variation in dietary Cd intake was primarily due to differences in the intake through boiled rice. Despite the recent reduction in Cd exposure, the current exposure level for Japanese people is still higher than the levels among other rice-dependent populations in Asia as well as in other parts of the world. Comparison was made between the present findings in general populations and observations among known Cd-pollution cases in Japan. CONCLUSIONS: Dietary uptake is an almost exclusive route of Cd exposure in the general Japanese population. Boiled rice is a strong determinant of variation in dietary Cd intake. Whereas there was a substantial reduction in Cd exposure among Japanese populations in the last 15 years, the current level is still high when compared internationally. PMID- 10672489 TI - Lumbar spine loads during education and training with self-contained breathing apparatus. AB - OBJECTIVE: German fire fighters often complain of lumbar back pain after training with a self-contained breathing apparatus while using a device called 'Schlaghammer' (SH). We investigated whether this training produces an excessive load on the lumbar spine. METHOD: We developed a model to estimate the load on the lumbar spine using a vector model similar to Jager et al.'s model. The force at the SH's cable was recorded on-line, and the position of each subject was registered with synchronized series of photos. The influence of the different parameters was calculated by varying these data (+/- 10%) and computing the model. RESULTS: There is no dangerous load for healthy persons using a correct technique. Improper technique, however, caused by exhaustion or ignorance, can result in high peaks of lumbar spine load. CONCLUSIONS: The lumbar spine loads involved are acceptable if the exercise is performed with a straight back without bending of the lumbar spine and without sudden acceleration. The direction of the pull should be towards the lumbar spine. PMID- 10672490 TI - The mechanism of a human reaction to vibration stress by palmar sweating in relation to autonomic nerve tone. AB - OBJECTIVES: To clarify the mechanism of a human reaction to vibration stress by palmar sweating in relation to the autonomic nerve tone. METHODS: The autonomic nerve tone was divided into four types by using digital photoelectroplethysmography (PTG) with auditory stimuli: normal (N), hyperreactive (I and D), and hyporeactive (P) types. Palmar sweating and digital PTG were simultaneously measured on the right palm and middle finger, respectively, in 20 healthy men. The left hand gripping the handle with a grasp strength of 49 N was exposed to vibration at a frequency of 125 Hz and acceleration magnitudes of 0 m/s2 (as a control), 30 m/s2, or 50 m/s2 for 3 min. The volume of palmar sweating was recorded before, during, and 30 min after vibration load. Three kinds of drugs related to the autonomic nervous system were orally administered to the subjects. Then 80 min after administration, the experiments were repeated. RESULTS: Of 20 subjects, 17 showed normal autonomic nerve tone (N type), and 3 hyperreactive (I type). The palmar sweating reaction to vibration in I-type subjects was greater and lasted longer than that in N-type subjects. Vibration with an acceleration of 50 m/s2 produced the greatest reaction which was about 7 times larger than that at 0 m/s2 and 2.5 times that at 30 m/s2 (P < 0.01). Sulpiride decreased palmar sweating during vibration, while prazosin and scopolamine inhibited it. CONCLUSIONS: The palmar sweating reaction to vibration stress was related to the background level of the autonomic nerve tone. The sweating volume was in direct proportion to the acceleration magnitude of vibration. The reaction of palmar sweating to vibration stress may be mediated through both the adrenergic and cholinergic fibers of the autonomic nervous system. PMID- 10672491 TI - Questionnaire surveys on health and working conditions: development of an instrument for risk assessment in companies. AB - Periodic Occupational Health Surveys (POHS) are frequently used by occupational health and safety services in the Netherlands as a risk assessment instrument. These surveys include a questionnaire on work and health. Systematic attention is paid in this questionnaire to a broad range of working conditions and health complaints. In this article a method is presented to identify and evaluate work risks and health problems in groups of workers. Working conditions and health in any given company or department are assessed by comparing questionnaire data from its worker populations with data from one or more reference populations. Significant differences are interpreted as signals for both adverse working conditions and health problems. Considerations and choices with regard to the technical, operational and strategic quality of the method are elucidated. Probabilities of alpha- and beta-errors, choice of significance levels, and selection of reference populations are dealt with. Finally, a way of presentation of the results is shown. The method is considered to be part of a broader approach toward risk assessment. We recommend the combined use of questionnaire results and other available information, such as workplace surveys and sickness absence data. Questionnaires about work and health can be seen as one step in a multi-phase design: like in many diagnostic processes, the latter phases can enhance the precision of previous results. Recommendations are made for validating and evaluating this instrument. PMID- 10672492 TI - Evaluation of half-mask respirator protection in styrene-exposed workers. AB - OBJECTIVE: The protection afforded by respirators to styrene (St)-exposed workers varies considerably. Our objective was to study the effective 'in the field' reduction in St exposure obtained by negative-pressure half-mask respirators worn by a group of fiberglass-reinforced plastics (FRP) workers. Protection was evaluated by measuring the reduction in urinary St (StU) excretion. METHODS: Seven FRP workers not using respiratory protection devices were studied for a week. External exposure to St was evaluated by personal passive sampling, and the internal dose by StU measurement. Then workers were asked to use a half-mask respirator for a week for the entire morning half-shift, and St exposure and internal dose were re-assessed. RESULTS: Mean environmental levels of St during the morning half-shift were 230-280 mg/m3, i.e., about three times the current limit proposed by ACGIH; the difference among days was not significant. Using respirators was accompanied by a large inter-individual and also intra-individual variability: the estimated reduction of StU values ranged from 30% to 90% (mean 60%). Mean StU values increased by 50% from Monday to Friday, while environmental St concentrations remained steady. Furthermore, the proportion of workers exceeding the biological equivalent exposure limit (BEEL) was 14% on Monday, double (33%) on Thursday, and triple (43%) on Friday. These data suggest a decrease of protection during the week. CONCLUSIONS: The protection afforded by negative-pressure half-mask respirators varies widely, which stresses the need to assess the effective reduction of exposure whenever these devices are introduced for St-exposed workers. If respirators are to be re-used for several days, their performance must be evaluated during the last shift of use. Measurement of urinary excretion of unmodified St proved a useful tool for the evaluation of respirator effectiveness in exposed workers. PMID- 10672493 TI - Aluminium dust-induced lung disease in the pyro-powder-producing industry: detection by high-resolution computed tomography. AB - OBJECTIVE: The aim of this case study was to investigate the suitability of high resolution computed tomography (HRCT) for detecting early stages of lung fibrosis induced by aluminium (Al) dust. METHODS: A 40-year-old worker was studied who had worked as a stamper for 14 years in a plant producing aluminium powder and had been exposed to high levels of aluminium dust during this time. The investigation included the collection of general data on health and details on occupational history, immunological tests, a physical examination, lung function analysis, biological monitoring of Al in plasma and urine, chest X-rays and HRCT. RESULTS: For many years the man has suffered from an exercise-induced shortness of breath. Lung function analysis revealed a reduction of the vital capacity to 57.5% of the predicted value. The Al concentration in plasma was 41.0 micrograms/l (upper reference value 10 micrograms/l) and in urine 407.4 micrograms/l upper reference value 15 micrograms/l, biological tolerance (BAT) value 200 micrograms/l[ at the time of diagnosis. Chest X-ray showed unspecific changes. HRCT findings were characterised by small, centrilobular, nodular opacities and slightly thickened interlobular septae. Exposure to other fibrotic agents could be excluded. CONCLUSIONS: HRCT was more sensitive than chest X-rays for detecting this early stage of Al-dust-induced lung disease. The suitability of HRCT in the surveillance of workers highly exposed to aluminium powder should be evaluated in further studies. Biological monitoring can be used to define workers at high risk. PMID- 10672494 TI - Urinary nickel as bioindicator of workers' Ni exposure in a galvanizing plant in Brazil. AB - OBJECTIVES: We measured urinary nickel (U-Ni) in ten workers (97 samples) from a galvanizing plant that uses nickel sulfate, and in ten control subjects (55 samples) to examine the association between occupational exposure to airborne Ni and Ni absorption. METHODS: Samples from the exposed group were taken before and after the work shift on 5 successive workdays. At the same time airborne Ni (A Ni) was measured using personal samplers. Ni levels in biological material and in the airborne were determined by a graphite furnace atomic absorption spectrometry validated method. In the control group the urine samples were collected twice a day, in the before and after the work shift, on 3 successive days. RESULTS: Ni exposure low to moderate was detected in all the examined places in the plant, the airborne levels varying between 2.8 and 116.7 micrograms/m3 and the urine levels, from samples taken postshift, between 4.5 and 43.2 micrograms/g creatinine (mean 14.7 micrograms/g creatinine). Significant differences in U-Ni creatinine were seen between the exposed and control groups (Student's t test, P < or = 0.01). A significant correlation between U-Ni and A-Ni (r = 0.96; P < or = 0.001) was detected. No statistical difference was observed in U-Ni collected from exposed workers in the 5 successive days, but significant difference was observed between pre- and postshift samples. CONCLUSIONS: Urinary nickel may be used as a reliable internal dose bioindicator in biological monitoring of workers exposed to Ni sulfate in galvanizing plants regardless of the day of the workweek on which the samples are collected. PMID- 10672495 TI - Ribosomal proteins in cell proliferation and apoptosis. AB - Ribosomal proteins have the complex task of coordinating protein biosynthesis to maintain cell homeostasis and survival. Recent evidence suggests that a number of ribosomal proteins have secondary functions independent of their involvement in protein biosynthesis. A number of these proteins function as cell proliferation regulators and in some instances as inducers of cell death. Specifically, expression of human ribosomal protein L13a has been shown to induce apoptosis, presumably by arresting cell growth in the G2/M phase of the cell cycle. In addition, inhibition of expression of L13a induces apoptosis in target cells, suggesting that this protein is necessary for cell survival. Similar results have been obtained in the yeast Saccharomyces cerevisiae, where inactivation of the yeast homologues of L13a, rp22 and rp23, by homologous recombination results in severe growth retardation and death. In addition, a closely related ribosomal protein, L7, arrests cells in G1 and also induces apoptosis. Thus, it appears that a group of ribosomal proteins may function as cell cycle checkpoints and compose a new family of cell proliferation regulators. PMID- 10672496 TI - The immunomodulatory role of CD4-positive cytotoxic T-lymphocytes in health and disease. AB - Among the CD4-positive (CD4+) T-lymphocytes a population exists which expresses cytolytic activity. These 'killer' cells belong to the T helper type 1 (Th1) subset and if activated, express Fas-ligand (FasL) which induces apoptosis in Fas positive target cells. The major targets of these CD4+ cytotoxic T-lymphocytes (CTL) are cells of the immune system, such as T, B cells and macrophages which express Fas upon activation. Thus, CD4+ CTL play a major immunoregulatory part through the elimination of activated myeloid and lymphoid cells during and upon completion of an immune response. In certain diseases, such as in HIV-infection and some autoimmune disorders, the functional activity of CD4+ CTL is disturbed preferentially at the level of FasL-Fas interaction, further emphasizing their important immunoregulatory role. Furthermore, Fas-ligand expressing tumors can evade the attack of Fas-positive CD4+ CTL and other effector cells, thereby giving them an opportunity to expand. PMID- 10672497 TI - Apoptosis of murine neonatal T cells. AB - Recent evidence supports the idea that T cells in neonatal animals are developmentally mature in their capacity to mount protective helper and cytotoxic responses. Nonetheless, neonates fall prey to infections which have little effect on adults and they often fail to mount mature responses to environmental, experimental, or vaccine antigens. To reconcile these observations, it may be important to consider the potential role of apoptosis in neonatal immune responses. In adults, apoptosis is well established as a centrally important process in the homeostasis of cellular immune responses. Activated T cells deprived of IL-2 undergo cytokine withdrawal-induced apoptosis. Previously activated T cells can also be triggered by secondary stimulation to undergo activation induced apoptosis. This review summarizes our current state of knowledge of apoptosis of murine neonatal T cells and discusses the possible impact(s) of this apoptosis on neonatal immune responses in vivo. PMID- 10672498 TI - Molecular and cellular mechanisms regulating T and B cell apoptosis through Fas/FasL interaction. AB - Fas (CD95) and Fas ligand (FasL) are a receptor/ligand pair critically involved in lymphocyte homeostasis and peripheral tolerance such that genetic defect in either Fas or FasL results in an autoimmune lymphoproliferative syndrome. Fas is a type I transmembrane protein and a member of the tumor necrosis factor receptor (TNFR) family whereas FasL is a type II transmembrane protein and a member of TNF family. Binding of Fas by FasL induces apoptosis of the Fas-expressing cells. In the past few years, Fas/FasL interaction has been connected to a series of important phenomena previously viewed as independent immune processes. The activation-induced T cell death (AICD) and the FasL-mediated cytotoxicity by activated T cells are two critical mechanisms that can account for most of these phenomena. It is in the context of the two mechanisms that we discuss in this review the molecular and cellular events that occur during T/T and T/B interactions that account for the down-regulation of the immune response. We have also discussed recent advances in the areas of FasL gene regulation, lymphokine regulation of AICD, and regulation of B cell susceptibility to FasL. Investigation in these areas should help elucidate the role of Fas/FasL in the complex network of regulatory mechanisms that control immune response and autoimmunity. PMID- 10672499 TI - Macrophage suppression of T cell activation: a potential mechanism of peripheral tolerance. AB - The mechanisms of induction and maintenance of tolerance in self-reactive T cells in the periphery are poorly understood. Current models assume that successful T cell activation only occurs if ligation of the T cell receptor (signal 1) by antigen presenting cells (APCs) is accompanied by a costimulatory signal (signal 2), and that signal 1 in the absence of signal 2 is either ignored or is tolerizing. However, there is also evidence for the existence of macrophages (M phi) capable of suppressing T cell activation both in vitro and in vivo. The possibility of a more actively induced tolerance exists, in which the M phi itself responds to T cell-mediated signals in a tolerogenic fashion. This would help to resolve the paradox that tissue M phi, which act as scavengers of self antigen, can also act as professional APCs. The ability of tissue macrophages to actively suppress T cells would further underscore the importance of the innate immune system in regulating adaptive immune responses. PMID- 10672500 TI - Involvement of Fas-Fas ligand interactions in graft rejection. AB - The Fas/Fas ligand (FasL) pathway has been shown to be important in T lymphocyte mediated cell death and is a key peripheral immunoregulatory mechanism that limits expansion of antigen-activated lymphocytes. The expression of Fas by commonly transplanted organs such as the heart, lung, kidney, and liver suggests that these tissues may be targets of FasL-expressing allospecific cytotoxic T lymphocytes. In this review the current literature examining the Fas/FasL system as a potential cellular effector pathway in tissue injury is discussed. In addition to a deleterious role in destruction of graft tissue, Fas/FasL interactions may have a beneficial role in transplantation. Recent studies suggest that modulation of FasL in target tissue leads to deletion, via apoptosis, of graft infiltrating lymphoid cells. However, an equally compelling series of reports indicate that overexpression of FasL can lead to a heightened immune response. These data are reviewed in the context of strategies to achieve long term allograft survival. PMID- 10672501 TI - Apoptosis and CD95 ligand in immune privileged sites. PMID- 10672502 TI - Apoptosis in human autoimmune diseases. AB - The description of apoptosis or programmed cell death nearly thirty years ago did not initially stimulate a great deal of interest. However, the ways cells die is clearly an essential part of biological homeostasis and well worth of study in its own right as the enormous literature on the subject in the past 15 years confirms. In the past decade new avenues of apoptosis research have opened up as the relationship between this form of cell death and autoimmune disease has come under increasing scrutiny. Although most research to date has been in animal study models, there are a variety of studies which have begun to explore links between apoptosis and a wider range of human autoimmune conditions. In this review we analyse briefly the background to what is known about apoptosis and focus on the increasing likelihood that abnormalities in apoptosis are contributory factors in the development of human autoimmunity. PMID- 10672503 TI - When immunization leads to autoimmunity: chronic inflammation as a result of thymic and mucosal dysregulation in IL-2 knock-out mice. PMID- 10672504 TI - Physician-hospital partnerships: incentive alignment through shared governance within a performance improvement structure. AB - BACKGROUND: Producing accessible, appropriate, and accountable medical care that improves the health of the populations served requires collaborative physician organization relationships within which performance measurement across the continuum of care occurs. Governance and shared responsibility for performance improvement (PI) through organizational structure and process have proven to be particularly complex challenges. REDESIGN OF THE PI SYSTEM: The Health Alliance of Central New York, based in Syracuse, New York, which consists in part of Crouse Hospital; ambulatory medical care sites and physicians; a physician organization, a physician-hospital organization, and an independent practice association; in February 1997 established a plan for a redesign of the PI system. IMPLEMENTING THE MODEL: In April 1998 the development of joint performance indicators, the Family of Measures, was undertaken. Recommendations for improvements necessary to correct process failures are referred to the medical staff executive committee and/or the appropriate coordinating committee, which then charges the appropriate service-line PI Council(s) with the responsibility for making those improvements. DISCUSSION: Systemwide PI with collaborative decision making by process stakeholders has been a major cultural transition requiring a degree of organizational readiness. Support of the most senior levels of management is critical. Institutional silos do not support shared, participatory decision making and cannot be overcome without strong support from senior management and in many cases the direct support and involvement of the CEO. Integrating information systems represents a considerable challenge: to find hardware and software that will interface properly to produce desired results, to successfully interface computer support personnel into the PI process, and to ensure the commitment to the financial resources to meet the information system requirements. In addition, meaningful and material reengineering requires substantial physician input. Simply reducing length of stay or cost per case is not an outcome that is by and large a strong motivator for physicians. Projects must have meaning at the level of the individual physician to raise interest and create buy-in. Enduring success will be achieved only through achievement of material and salient improvements (for both physicians and the institution) in combination with careful alignment of physician and institutional incentives. PMID- 10672505 TI - Continuous self-improvement: systems thinking in a personal context. AB - BACKGROUND: Continuous quality improvement (CQI) thinking and tools have broad applicability to improving people's lives--in continuous self-improvement (CSI). Examples include weight loss, weight gain, increasing exercise time, and improving relationship with spouse. In addition, change agents, who support and facilitate organizational efforts, can use CSI to help employees understand steps in CQI. A STEP-BY-STEP APPROACH: Team members should be involved in both the definition of the problem and the search for the solution. How do everyday processes and routines affect the habit that needs to change? What are the precursors of the event? Clients list possible solutions, prioritize them, and pilot test the items selected. One needs to change the daily routines until the desired behavior is accomplished habitually and with little external decision. DISCUSSION: CSI is successful because of its emphasis on habits embedded in personal processes. CSI organizes support from process owners, buddies, and coaches, and encourages regular measurement, multiple small improvement cycles, and public reporting. PMID- 10672506 TI - Does this patient need to be evaluated today? Designing a guideline-driven triage process to determine the timing of care for adults with respiratory infection symptoms. AB - BACKGROUND: Physicians and nurses often make judgments about the urgency with which patients require evaluation, yet few explicit process-of-care criteria are available to guide these decisions. Using a multidisciplinary expert physician panel and explicit, quantitative group judgment methods, standardized, clinically detailed deferred care criteria were developed to guide emergency department and ambulatory care triage decisions for same-day versus deferred care for patients with respiratory infection symptoms. METHODS: Using a modified Delphi process, an eight-member multidisciplinary expert physician panel rated the safety of deferred care for standardized clinical scenarios. The ratings were converted into explicit criteria and then compared with usual implicit judgment in terms of nurse triage times. RESULTS: The panel achieved 100% consensus on 36 critical clinical factors, each of which precludes deferring care for a patient with respiratory infection symptoms. Based on combinations of 12 additional clinical factors, 48 clinical scenarios were created that the panel rated for deferred care safety. Panelists' ratings agreed for 90% of clinical scenarios. These were formatted into screening criteria. Near-perfect interrater agreement (kappa = 0.9) was found in reproducibility testing. The difference in mean nurse triage times using the criteria compared with implicit nurse judgment was 0.4 minutes (95% confidence interval = -2.1 to 2.9 minutes). CONCLUSIONS: Application of explicit criteria for deferring care of patients with respiratory infection symptoms did not lengthen triage time. This approach may facilitate more efficient resource management for ambulatory settings. However, widespread use before these criteria's, our systematic criteria-based triage should be validated in multicenter clinical trials against an outcome standard and the more common implicit approach. PMID- 10672507 TI - Setting thresholds for quality indicators derived from MDS data for nursing home quality improvement reports: an update. AB - BACKGROUND: Determining meaningful thresholds to reinforce excellent performance and flag potential problem areas in nursing home care is critical for preparing reports for nursing homes to use in their quality improvement programs. This article builds on the work of an earlier panel of experts that set thresholds for quality indicators (QIs) derived from Minimum Data Set (MDS) assessment data. Thresholds were now set for the revised MDS 2.0 two-page quarterly form and Resource Utilization Groups III (RUGS III) quarterly instrument. SETTING THRESHOLDS: In a day-long session in October 1998, panel members individually determined lower (good) and upper (poor) threshold scores for each QI, reviewed statewide distributions of MDS QIs, and completed a follow-up Delphi of the final results. REPORTING MDS QIS FOR QUALITY IMPROVEMENT: The QI reports compiled longitudinal data for all residents in the nursing home during each quarter and cumulatively displayed data for five quarters for each QI. A resident roster was provided to the nursing home so that the quality improvement team could identify the specific residents who developed the problems defined by each QI during the last quarter. Quality improvement teams found the reports helpful and easy to interpret. SUMMARY AND CONCLUSIONS: As promised in an earlier report, to ensure that thresholds reflect current practice, research using experts in a panel to set thresholds was repeated as needed. As the MDS instrument or recommended calculations for the MDS QIs change, thresholds will be reestablished to ensure a fit with the instrument and data. PMID- 10672508 TI - Periodic solutions: a robust numerical method for an S-I-R model of epidemics. AB - We describe and analyze a numerical method for an S-I-R type epidemic model. We prove that it is unconditionally convergent and that solutions it produces share many qualitative and quantitative properties of the solution of the differential problem being approximated. Finally, we establish explicit sufficient conditions for the unique endemic steady state of the system to be unstable and we use our numerical algorithm to approximate the solution in such cases and discover that it can be periodic, just as suggested by the instability of the endemic steady state. PMID- 10672509 TI - Transport effects on surface-volume biological reactions. AB - Many cellular reactions involve a reactant in solution binding to or dissociating from a reactant confined to a surface. This is true as well for a BIAcore, an optical biosensor that is widely used to study the interaction of biomolecules. In the flow cell of this instrument, one of the reactants is immobilized on a flat sensor surface while the other reactant flows past the surface. Both diffusion and convection play important roles in bringing the reactants into contact. Usually BIAcore binding data are analyzed using well known expressions that are valid only in the reaction-limited case when the Damkohler number Da is small. Asymptotic and singular perturbation techniques are used to analyze dissociation of the bound state when Da is small and O(1). Linear and nonlinear integral equations result from the analysis; explicit and asymptotic solutions are constructed for physically realizable cases. In addition, effective rate constants are derived that illustrate the effects of transport on the measured rate constants. All these expressions provide a direct way to estimate the rate constants from BIAcore binding data. PMID- 10672511 TI - Measured and modeled properties of mammalian skeletal muscle. II. The effects of stimulus frequency on force-length and force-velocity relationships. AB - Interactions between physiological stimulus frequencies, fascicle lengths and velocities were analyzed in feline caudofemoralis (CF), a hindlimb skeletal muscle composed exclusively of fast-twitch fibers. Split ventral roots were stimulated asynchronously to produce smooth contractions at sub-tetanic stimulus frequencies. As described previously, the peak of the sub-tetanic force-length relationship was found to shift to longer lengths with decreases in stimulus frequency, indicating a length dependence for activation that is independent of filament overlap. The sub-tetanic force-velocity (FV) relationship was affected strongly both by stimulus frequency and by length; decreases in either decreased the slope of the FV relationship around isometric. The shapes of the force transients following stretch or shortening revealed that these effects were not due to a change in the instantaneous FV relationship; the relative shape of the force transients following stretch or shortening was independent of stimulus frequency and hardly affected by length. The effects of stimulus frequency and length on the sub-tetanic FV relationship instead appear to be caused by a time delay in the length-dependent changes of activation. In contrast to feline soleus muscle, which is composed exclusively of slow-twitch fibers, CF did not yield at sub-tetanic stimulus frequencies for the range of stretch velocities tested (up to 2 L0/s). The data presented here were used to build a model of muscle that accounted well for all of the effects described. We extended our model to account for slow twitch muscle by comparing our fast-twitch model with previously published data and then changing the necessary parameters to fit the data. Our slow-twitch model accounts well for all previous findings including that of yielding. PMID- 10672512 TI - Troponin C regulates the rate constant for the dissociation of force-generating myosin cross-bridges in cardiac muscle. AB - It is well known that cardiac troponin C (cTnC) regulates the association of force-generating myosin cross-bridges. We report here evidence for an additional role for cTnC. This hypothesis states that Ca2+ binds more strongly to cTnC when force-generating myosin cross-bridges are attached to actin and that removal of this bound Ca2+ accelerates the dissociation of force-generating myosin cross bridges. Intact Fura-2-loaded rat papillary muscles and skinned (permeabilized) ventricular preparations were used. The preparations were mounted in the Guth Muscle Research System which is capable of measuring simultaneously fluorescence and force in response to length perturbations. All mechanical perturbations of muscle length (isotonic shortening, quick stretches and releases, and length vibrations) which cause dissociation of force-generating myosin cross-bridges during a twitch resulted in Ca2+ being released from troponin as judged from changes in the Ca2+ transients (Fura-2 (340/380) fluorescence ratio). Thus dissociation of force-generating myosin cross-bridges cause Ca2+ to be released from cTnC. Conversely, it would be expected that removal of strongly bound Ca2+ from cTnC would result in an increase in the rate of dissociation of force generating myosin cross-bridges. To test this hypothesis actomyosin ATPase (NADH fluorescence change) and isometric force were measured in skinned cardiac preparations. The ratio of the ATPase/Force is proportional to the rate constant (gapp) for the dissociation of force-generating myosin cross-bridges. The data showed that decreasing the amount of Ca2+ bound to cTnC in skinned cardiac fibers caused an increase in the ratio of ATPase/Force, the rate of dissociation (gapp) of force-generating myosin cross-bridges. PMID- 10672513 TI - The structure of the avian fast skeletal muscle troponin T gene: seven novel tandem-arranged exons in the exon x region. AB - To elucidate the mechanism that produces enormous molecular diversity in troponin T (TnT) of fast skeletal muscle, we determined the 5'-half genomic sequence of the chicken fast muscle TnT gene. The sequence of ca. 16 kb included seven exons (exons 1, 2, 3, 4, w, 5, and 6), which have been reported previously and presumed by sequencing TnT cDNAs. Additionally we found six 15 nt and one 18 nt sequences in the region between exons 5 and 6 (i.e. the exon x region). They were encompassed by consensus splice donor and acceptor sites and preceded by putative branch sites, and designated herein as exons xa to xg. Our result shows that the sequence derived from exons x1, x2, and x3, the exons presumed previously by cDNA sequencing, is actually encoded by the seven exons xa to xg, establishing the precise gene structure in the exon x region. Based on our data, together with that on the 3'-half genomic sequence of the quail fast muscle TnT gene, we conclude that the avian fast skeletal muscle TnT gene includes 27 exons, 16 of which are alternatively spliced. PMID- 10672510 TI - Understanding dystrophinopathies: an inventory of the structural and functional consequences of the absence of dystrophin in muscles of the mdx mouse. PMID- 10672514 TI - Nspl1, a new Z-band-associated protein. AB - Molecular characterization of a novel gene designated Neuroendocrine-Specific Protein-Like-1 (Nspl1) had revealed that this gene is expressed as two transcripts, a 1.2 kb transcript found predominantly in skeletal muscle and a 2.1 kb transcript expressed in the brain. The exceptionally high level of skeletal muscle expression prompted us to determine where the protein is localized to skeletal muscle. In vitro studies were performed using two plasmid constructs that generate full-length Nspl1 muscle-specific protein fused to the green fluorescent protein (GFP). In one construct, the GFP cDNA was fused to the N terminus of the Nspl1 cDNA while in the second construct, the GFP cDNA was fused to the C-terminus of the Nspl1 cDNA. Transfection of either plasmid into mononucleated myoblasts showed that the Nspl1-GFP chimeric protein was associated with intermediate filaments. This was confirmed by using an antibody to stain desmin and finding that GFP-Nspl1 colocalizes with desmin. Chick primary myoblasts were transfected with the chimeric cDNAs and allowed to differentiate into mature myotubes. Results from this analysis and the use of monoclonal antibody to stain alpha-actinin, further localized the Nspl1 protein to the Z band of mature myotubes. Confocal microscopy of the myotubes containing Nspl1-GFP demonstrates that Nspl1 is distributed continuously throughout the Z-disks. PMID- 10672515 TI - Expression and localisation of annexin VII (synexin) isoforms in differentiating myoblasts. AB - Annexin VII exists in a 47 kDa and a 51 kDa isoform with the 51 kDa protein being the only isoform present in skeletal muscle. Expression of the 51 kDa isoform during myogenesis and localization was studied in cells after conversion into myogenic cells by transduction with MyoD and in mouse and human myogenic cell lines. MyoD expression in NIH3T3 and C3H10T1/2 fibroblasts led to disappearance of the mRNA specific for the 47 kDa isoform and appearance of the 51 kDa isoform specific mRNA. The overall amount of annexin VII protein was reduced in myogenic converted cells. Both in undifferentiated and differentiated cells annexin VII was localized by immunofluorescence microscopy to punctate structures which were distributed all over the cell. A GFP annexin VII fusion protein showed a similar distribution. Cell fractionation studies indicated that annexin VII is equally distributed between cytosol and membrane fractions in undifferentiated cells, while in differentiated cells it is exclusively present in the membrane fraction. By sucrose gradient centrifugation of postnuclear supernatants we identified two distinct annexin VII-containing membrane populations that cofractionated with caveolin 3- and sorcin-containing membranes. PMID- 10672516 TI - Differences in the profile of unfused tetani of fast motor units with respect to their resistance to fatigue in the rat medial gastrocnemius muscle. AB - In most studies performed on motor units in mammalian muscles the division of these units into fast and slow types has been based on the 'sag' visible in the profile of unfused tetanus. The time course of the sag in unfused tetani of fast motor units was analysed in the present study. Fast units of rat medial gastrocnemius muscle were classified as fast fatigable (FF) or fast resistant to fatigue (FR) on the basis of a fatigue index calculated during the standard fatigue test. In middle-fused tetani (fusion index 0.25-0.75), it was observed that for FF motor units the sag was shorter and occurred earlier than for FR units. Moreover, in FF units, the sag was followed by potentiating tension, whereas for FR units this potentiation was weaker or even absent. A tetanus shape index, which expressed the ratio of the area of the first part of the tetanus record (between the tension record and the baseline, from the beginning of tetanus up to the lowest point during the sag in the tension record) to the area under the second part of tetanus (from this lowest point up to the end of the record) was introduced. For FF units, this index ranged from 0.13 to 0.47, whereas for FR units it ranged from 0.54 to 17.8 (with one exception). These results showed that the difference in unfused tetanus expressed in this tetanus shape index could be used as an accurate alternative method of dividing fast units into FF and FR groups. Moreover, the difference in sag time course in FF and FR groups. Moreover, the difference in sag time course in FF and FR units suggests that the metabolism responsible for this contractile phenomenon is significantly different time courses in IIa and IIb muscle fibres, constituting FF and FR units, respectively. PMID- 10672517 TI - An electrophoretic study of myosin heavy chain expression in skeletal muscles of the toad Bufo marinus. AB - In this study we developed an SDS-PAGE protocol which for the first time separates effectively all myosin heavy chain (MHC) isoforms expected to be expressed in iliofibularis (IF), pyriformis (PYR), cruralis (CRU) and sartorius (SAR) muscles of the toad Bufo marinus on the basis of previously reported fibre type composition. The main feature of the method is the use of alanine instead of glycine both in the separating gel and in the running buffer. The correlation between the MHC isoform composition of IF, SAR and PYR muscles determined in this study and the previously reported fibre type composition of IF and SAR muscles in the toad and of PYR muscle in the frog was used to tentatively identify the MHC isoforms expressed by twitch fibre types 1, 2 and 3 and by tonic fibres. The alanine-SDS electrophoretic method was employed to examine changes in the MHC composition of IF, PYR, CRU and SAR muscles with the ontogenetic growth of the toad from post-natal life (body weight < 1 g) to late adulthood (body weight 200 450 g). The developmental changes in the MHC isoform composition of the toad IF muscle observed in this study are in very good agreement with those in the fibre type composition of the developing IF muscle reported in the literature. PMID- 10672518 TI - Junctions between subsynaptic folds and rough sarcoplasmic reticulum of muscle fibres. AB - Serial sections through motor end plate regions of mouse muscle fibres demonstrated junctions between the subsynaptic folds and the rough sarcoplasmic reticulum of the sole plate nuclei. The shape of these structures resembles that of the well-known peripheral couplings, diads and triads of muscle fibres. However, the location of the new junctions between the surface membrane and the sole plate nuclei at a large distance from myofibrils, indicates a different function. The connection with the rough sarcoplasmic reticulum possibly influence the regulation of fibre protein metabolism, for example, gene expression of acetylcholine receptor synthesis. PMID- 10672519 TI - The interface between MyBP-C and myosin: site-directed mutagenesis of the CX myosin-binding domain of MyBP-C. AB - Myosin-binding protein-C (MyBP-C or C-protein) is a ca. 130 kDa protein present in the thick filaments of all vertebrate striated muscle. The protein contains ten domains, each of ca. 90-100 amino acids; seven are members of the IgI family of proteins, three of the fibronectin type III family. The motifs are arranged in the following order (from N- to C-terminus): Ig-Ig-Ig-Ig-Ig-Fn-Fn-Ig-Fn-Ig. The C terminal Ig motif (domain X or CX) contains its light meromyosin-binding site. A recombinant form of CX, beginning at Met-1027, exhibits saturable binding to myosin with an affinity comparable to the C-terminal 13 kDa chymotryptic fragment of native MyBP-C. To identify the surface in CX involved in its interaction with myosin, nine site-directed mutants (R37E, K43E, N49D, E52R, D56K, R73E, R74E, G80D and R103E) were constructed. Using a new assay for assessing the binding of CX with the light meromyosin (LMM) portion of myosin, we demonstrate that recombinant CX, just as the full-length protein, is able to facilitate LMM polymerization. Moreover, we show that residues Arg-37, Glu-52, Asp-56, Arg-73, and Arg-74 are involved in this interaction with the myosin rod. All of these amino acids interact with negatively charged residues of LMM, since the mutants R37E, R73E and R74E are unable to bind myosin, whereas E52R and D56K bind myosin with higher affinity than wild-type CX. Residues Lys-43 and Arg-103 show a small but significant influence on the binding reaction; residues Asn-49 and Gly-80 seem not to be involved in this interaction. Based on these data, a model is proposed for the interaction between MyBP-C CX and myosin filaments. In this model, CX interacts with four molecules of LMM at four different sites of the binding protein, thus explaining the effects of MyBP-C on the critical concentration of myosin polymerization. PMID- 10672520 TI - Comparison of cross-bridge cycling kinetics in neonatal vs. adult rat ventricular muscle. AB - The developmental shift in contractile protein isoform expression in the rodent heart likely affects actin-myosin cross-bridge interactions. We compared the Ca2+ sensitivity for force generation and cross-bridge cycling kinetics in neonatal (postnatal days 0-3) and adult (day 84) rats. The force-pCa relationship was determined in Triton-X skinned muscle bundles activated at pCa 9.0 to 4.0. In strips maximally activated at pCa 4.0, the following parameters of cross-bridge cycling were measured: (1) rate of force redevelopment following rapid shortening and restretching (ktr); and (2) isometric stiffness at maximal activation and in rigor. The fraction of attached cross-bridges (alpha fs) and apparent rate constants for cross-bridge attachment (fapp) and detachment (gapp) were derived assuming a two-state model for cross-bridge cycling. Compared to the adult, the force-pCa curve for neonatal cardiac muscle was significantly shifted to the left. Neonatal cardiac muscle also displayed significantly smaller alpha fs, slower ktr and fapp; however, gapp was not significantly different between age groups. These data indicate that weaker force production in neonatal cardiac muscle involves, at least in part, less efficient cross-bridge cycling kinetics. PMID- 10672521 TI - A note on the caldesmon sequence. PMID- 10672522 TI - Measurement of UV radiation using suspensions of microorganisms. AB - The measurement of solar UV radiation is usually performed using physical devices like photodiodes or photomultipliers or with chemical substances (actinometry). The application of biological material such as microorganisms for this purpose has gained increasing importance in the last few years. The microorganisms may be dried and spread on a flat surface or they may be in aqueous suspensions contained in UV-transparent vessels. If the measurements are done on flat surfaces, the irradiance weighted by the action spectrum of the dried microorganism used is the result of the measurement. If aqueous suspensions of microorganisms are used, contained for instance in spherical vessels, the fluence weighted by the action spectrum of the microorganisms in the aqueous suspension is the result. A problem of this method of measurement can be that inside the vessel the distribution of UV radiation is usually not homogeneous, causing distributions of fluences among the irradiated microorganisms, which may result in variation of the results depending on the mixing characteristics of the suspension during irradiation. PMID- 10672523 TI - Comparative measurements of solar UV radiation with spore dosimetry at three European and two Japanese sites. AB - Spore dosimetry of solar UV radiation has been employed in several field intercomparison campaigns carried out in summer months in Japan and Europe. The dose-rate profiles, total daily doses and personal daily exposures have been determined and compared at five sites based on the spore inactivation dose (SID). The maximum dose rate (0.83 SID/min) and the maximum daily dose (197 SID) observed at subtropical Naha (26.2 degrees N) are about three times those observed at subarctic Abisko (68.4 degrees N). The amounts of personal exposure during three European campaigns are moderate and show a tendency of inverse relationships with the daily doses. PMID- 10672524 TI - 'Vitamin D' biodosimeter: basic characteristics and potential applications. AB - The biologically important process of endogenous synthesis of vitamin D under UV solar irradiation is widespread in the biosphere and inherent to most animals and plants. A new method of biological dosimetry of UV radiation based on an in vitro model of vitamin D synthesis ('D-dosimeter') is discussed. Unlike the vast majority of biodosimeters, the action of which depends on the UV sensitivity of DNA and thus reflects damaging effects of UV radiation, the process of vitamin D synthesis is beneficial by its nature. To date, the complex network of photo- and thermoreactions of vitamin D synthesis in vitro is well understood, and an adequate mathematical model is available, ensuring a link between biological and physical units. Original spectral analysis of the multicomponent photoisomer mixture has been specially designed to provide the most effective use of the D dosimeter in situ. Spectral selectivity (exceptional sensitivity of certain parameters to the spectral composition of UV radiation) extends the usefulness of the method. PMID- 10672525 TI - The role of the spectral sensitivity curve in the selection of relevant biological dosimeters for solar UV monitoring. AB - To estimate the risk of enhanced UV-B radiation due to stratospheric ozone depletion, phage T7 and uracil thin-layer biological dosimeters have been developed, which weight the UV irradiance according to induced DNA damage. To study the molecular basis of the biological effects observed after UV irradiation, the spectral sensitivity curves of the two dosimeters and induction of the two major DNA photoproducts, cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts ((6-4)PDs), in phage T7 have been determined for polychromatic UV sources. CPDs and (6-4)PDs are determined by lesion-specific monoclonal antibodies in an immunodotblot assay. Phage T7 and uracil biological dosimeters together with a Robertson-Berger (RB) meter have been used for monitoring environmental radiation from the polar region to the equator. The biologically effective dose (BED) established with the three different dosimeters increases according to the changes in the solar angle and ozone column, but the degree of the change differs significantly. The results can be explained based on the different spectral sensitivities of the dosimeters. A possible method for determining the trend of the increase in the biological risk due to ozone depletion is suggested. PMID- 10672526 TI - Influence of spectral and angular sensitivity on the readout of biological dosimeters. AB - Biological systems used as biological dosimeters can possess different angular sensitivities from the detectors usually used in physical devices. A simple experimental setup has been developed and used to measure the angular sensitivity of uracil thin-layer biological dosimeters. Results of angular sensitivity measurements for uracil thin-layer dosimeters are presented using a Xe arc lamp as the UV source. According to the experiments described here, uracil thin-layer dosimeters show a cosine-type angular dependence. In several indoor experiments broadband UV meters are used to control the applied dose rate from a given artificial UV source. The experimental setup has been designed and used to verify experimentally the importance of spectral and angular sensitivity differences of biological and physical UV meters applied in biological experiments. Model calculations for two different irradiation systems, using different geometrical arrangements of artificial UV sources, are also presented. For these arrangements relative dose rates that could be measured with dosimeters of arbitrary spectral, but different angular sensitivity have been calculated. PMID- 10672527 TI - Biological UV dosimeters in the assessment of the biological hazard from environmental radiation. AB - To determine the impact of environmental UV radiation, biological dosimeters that weight directly the incident UV components of sunlight have been developed, improved and evaluated in the frame of the BIODOS project. Four DNA-based biological dosimeters ((i) phage T7, (ii) uracil thin layer, (iii) spore dosimeter and (iv) DLR-biofilm) have been assessed from the viewpoint of their biological relevance, spectral response and quantification of their biological effectiveness. The biological dosimeters have been validated by comparing their readings with weighted spectroradiometer data, by comparison with other biological doses, as well as with the determined amounts of DNA UV photoproducts. The data presented here demonstrate that the biological dosimeters are potentially reliable field dosimeters for measuring the integrated biologically effective irradiance for DNA damage. PMID- 10672528 TI - Photodynamic parameters in the chick chorioallantoic membrane (CAM) bioassay for topically applied photosensitizers. AB - The relative efficacy of Photofrin-based photodynamic therapy (PDT) has been compared with that of the second-generation photosensitizers 5-aminolevulinic acid (ALA), sulfonated chloro-aluminum phthalocyanine (AlPcSn), benzoporphyrin derivative monoacid ring A (BPD-MA), and lutetium texaphyrin (Lutex). PDT-induced vascular damage in the chick chorioallantoic membrane (CAM) is measured following topical application of the photosensitizers. In order to make meaningful comparisons, care is taken to keep treatment variables the same. These include light dose (5 and 10 J/cm2), power density (33 and 100 mW/cm2), and drug uptake time (30 and 90 min). The drug dose ranges from 0.1 microgram/cm2 for BPD to 5000 micrograms/cm2 for ALA. Results are also analyzed statistically according to CAM vessel type (arterioles versus venules), vessel diameter, and vessel development (embryonic age). For each photosensitizer, the order of importance for the various PDT parameters is found to be unique. The differences between the sensitizers are most likely due to variation in biophysical and biochemical characteristics, biodistribution, and uptake kinetics. PMID- 10672529 TI - Comparison of phototoxicity mechanism between pulsed and continuous wave irradiation in photodynamic therapy. AB - A study has been conducted in which HeLa cells are incubated with hematoporphyrin derivative (HpD) for 1 h (1 microgram/ml of HpD in PBS) to compare the use of continuous wave (CW) and pulsed laser (10 Hz repetition rate and 7-9 ns pulse width) light for photodynamic therapy. Cytotoxic effects on the cells are evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2-5-diphenyl-2H-tetrazolium bromide (MTT) assay and the fluorescein diacetate (FDA)/propidium iodide (PI) stain method using a flow cytometer. The type of cell death is estimated by analysis of the DNA content and observation of the nuclear morphology. The cytotoxicity ratio of cells irradiated by pulsed laser light is estimated to be lower than that for CW laser light. The viability of cells that received pulsed laser light gradually decreases, whereas no significant changes are found in the cells irradiated with CW laser light with the elapse of post-irradiation time. The type of cell death differs between the pulsed and CW laser light irradiations. These findings suggest that the cytotoxic efficacy of the excitation light source is displayed by the difference in the type of cell death, namely apoptosis or necrosis. PMID- 10672530 TI - A theoretical and experimental study of two thiazole orange derivatives with single- and double-stranded oligonucleotides, polydeoxyribonucleotides and DNA. AB - The effect of interaction with DNA and oligonucleotides on the photophysical properties of two thiazole orange (TO) derivatives, with different side chains ( (CH2)3-N+(CH3)3 and -(CH2)6-I)) linked to the nitrogen of the quinoline ring of the thiazole orange, is presented here. The first one called TO-PRO1 is a commercially available dye, whereas the second one called TO-MET has been specially synthesized for further covalent binding to oligonucleotides with the aim of being used for specific in situ detection of biomolecular interactions. Both photophysical measurements and molecular calculations have been done to assess their possible mode of interaction with DNA. When dissolved in buffered aqueous solutions both derivatives exhibit very low fluorescence quantum yields of 8 x 10(-5) and 2 x 10(-4), respectively. However, upon binding to double stranded DNA, large spectroscopic changes result and the quantum yield of fluorescence is enhanced by four orders of magnitude, reaching values up to phi F = 0.2 and 0.3, respectively, as a result of an intercalation mechanism between DNA base pairs. A modulation of the quantum yield is observed as a function of the base sequence. The two derivatives also bind with single-stranded oligonucleotides, but the fluorescence quantum yield is not so great as that when bound to double-stranded samples. Typical fluorescence quantum yields of 7 x 10( 3) to 3 x 10(-2) are observed when the dyes interact with short oligonucleotides, whereas the fluorescence quantum yield remains below 10(-2) when interacting with single-stranded oligonucleotides. This slight but significant quantum-yield increase is interpreted as a folding of the single strand around the dye, which reduces the internal rotation of the two heterocycles around the central methine bridge that links the two moieties of the dye. From these properties, it is proposed to link monomer covalently to oligonucleotides for the subsequent detection of target sequences within cells. PMID- 10672531 TI - Horizontal and sun-normal spectral biologically effective ultraviolet irradiances. AB - The dependence of the spectral biologically effective solar UV irradiance on the orientation of the receiver with respect to the sun has been determined for relatively cloud-free days at a sub-tropical Southern Hemisphere latitude for the solar zenith angle range 35-64 degrees. For the UV and biologically effective irradiances, the sun-normal to horizontal ratio for the total UV ranges from 1.18 +/- 0.05 to 1.27 +/- 0.06. The sun-normal to horizontal ratio for biologically effective irradiance is dependent on the relative effectiveness of the relevant action spectrum in the UV-A waveband. In contrast to the total UV, the diffuse UV and diffuse biologically effective irradiances are reduced in a sun-normal compared with a horizontal orientation by a factor ranging from 0.70 +/- 0.05 to 0.76 +/- 0.03. PMID- 10672532 TI - Photodynamic therapy using 5-aminolaevulinic acid for oesophageal adenocarcinoma associated with Barrett's metaplasia. AB - Photodynamic therapy (PDT) is a novel technique for local endoscopic treatment of gastrointestinal neoplasia. Current photosensitisers for PDT may cause prolonged skin phototoxicity. 5-Aminolaevulinic acid (ALA), a precursor of the photosensitiser protoporphyrin IX (PpIX), is more acceptable because of its short half-life and preferential accumulation in mucosa and mucosal tumour. We have treated 12 patients, median age 73 years (range 55-88) with oesophageal adenocarcinoma arising from Barrett's metaplasia (two carcinomas-in-situ, grade 0; 10 carcinomas, grade 1-11A based on endoluminal ultrasound in two and CT scanning in 10 patients). ALA (60 and 75 mg/kg body weight) was given orally in two or five equally divided doses. The PpIX distribution in stomach, normal oesophagus, Barrett's mucosa and carcinoma was measured by quantitative fluorescence photometry. PDT was performed using laser light (630 nm) delivered via a cylindrical diffuser 4-6 h after the first dose of ALA. The patients received one to four sessions of PDT. PpIX accumulation in the mucosa was two to three times that in the lamina propria. The differential distribution between carcinomatous and normal oesophageal mucosa was less marked (carcinoma:normal mucosa ratio = 1.4). Higher doses of ALA increased PpIX accumulation in all tissues but did not increase the differential PpIX distribution between tumour and normal oesophageal mucosa. After PDT using ALA (ALA/PDT), all mucosa showed superficial white necrotic changes and the histology confirmed fibrinoid necrosis. One patient with carcinoma-in-situ had the tumour eradicated after one treatment with no recurrence at 28 months. Another patient with a small T1 tumour required four ALA/PDT treatments, and died of other disease after 36 months. There was no evidence of recurrence. The tumour bulk in the other carcinomas was not significantly reduced. ALA/PDT has a potential for the eradication of small tumours but careful patient selection with endoluminal ultrasound is needed when using ALA/PDT to treat oesophageal cancer. PMID- 10672533 TI - Acid-base properties of chlorin e6: relation to cellular uptake. AB - Chlorins are attractive compounds for photodynamic therapy because of their high absorption in the red spectral region. In this study, the absorbance, fluorescence excitation and fluorescence emission spectra of chlorin e6 have been recorded as functions of pH in phosphate-buffered saline (PBS) solution with and without fetal calf serum (FCS). For pure PBS solutions, variation of the pH of the solution results in a shift of both the absorption and the fluorescence spectrum as well as in a decrease of the fluorescence intensity. Spectrophotometric and fluorimetric titration curves, based on observed changes, have been plotted. There is an indication of aggregate formation at low pH values (pH < 5). The presence of 5% FCS results in a shift of the titration curve, from an inflection point at about 6.5 to one at about 7.6. Pronounced spectral changes of the fluorescence emission spectra of protein-bound chlorin e6 (change of spectral shape, decrease of peak intensity) are also observed. The partition coefficients in the 1-octanol-water system increase with decreasing pH. Thus, relatively more of the drug is incorporated in the octanol phase at low pH. Cellular uptake of chlorin e6 in the presence of serum is significantly higher at pH 6.7 as compared with that at 7.3 and 7.6. We conclude that a change in the pH value of the surrounding medium leads to a change in the lipophilicity of chlorin e6. Such a change is likely to influence its binding to the serum proteins as well as its interaction with the plasma membrane of cells and may also be related to the selective tumor uptake of the drug. PMID- 10672534 TI - Synthesis, photochemistry and application of (7-methoxycoumarin-4-yl)methyl-caged 8-bromoadenosine cyclic 3',5'-monophosphate and 8-bromoguanosine cyclic 3',5' monophosphate photolyzed in the nanosecond time region. AB - New caged derivatives of hydrolysis-resistant 8-bromoadenosine cyclic 3',5' monophosphate (8-Br-cAMP) and 8-bromoguanosine cyclic 3',5'-monophosphate (8-Br cGMP) are described. The compounds are the axial and equatorial isomers of the (7 methoxycoumarin-4-yl)methyl (MCM) esters of cyclic nucleotides. Synthesis is accomplished by treatment of 4-bromomethyl-7-methoxycoumarin with the tetra-n butylammonium salts of the 8-bromo-substituted cyclic nucleotides or with the free acids of 8-Br-cAMP and 8-Br-cGMP in the presence of silver(I) oxide. MCM caged 8-Br-cAMP and MCM-caged 8-Br-cGMP liberate 8-Br-cAMP and 8-Br-cGMP during irradiation with ultraviolet light within a few nanoseconds. They show favorable absorption properties and quantum yields and are resistant to hydrolysis in aqueous buffer solutions. The moderate fluorescence properties of the caged compounds in comparison with the strongly fluorescent 4-hydroxymethyl-7 methoxycoumarin (MCM-OH) photoproduct allow the indirect estimation of the amount of photolytically released cyclic nucleotides in aqueous buffer solutions using fluorescence measurements. Their usefulness for physiological studies has been examined in a mammalian cell line expressing the cyclic nucleotide-gated ion channel of bovine olfactory sensory neurons using the patch-clamp technique and confocal laser scanning microscopy. The caged compounds serve as efficient and rapid intracellular sources of 8-Br-cAMP and 8-Br-cGMP. However, at least in HEK 293 cells, fluorescence signals cannot be used to monitor the photolysis of MCM caged 8-Br-cAMP and 8-Br-cGMP, due to quenching of the fluorescence of MCM-OH. PMID- 10672535 TI - Photothermal sensitization of amelanotic melanoma cells by Ni(II)-octabutoxy naphthalocyanine. AB - Incubation of B78H1 amelanotic melanoma cells with a potential photothermal sensitizer, namely, liposome-incorporated Ni(II)-octabutoxy-naphthalocyanine (NiNc), induces an appreciable cellular accumulation of the naphthalocyanine, which is dependent on both the NiNc concentration and the incubation time. No detectable decrease in cell survival occurs upon red-light irradiation (corresponding to the longest-wavelength absorption bands of NiNc) in a continuous-wave (c.w.) regime of the naphthalocyanine-loaded cells. On the other hand, 850 nm irradiation with a Q-switched Ti:sapphire laser operating in a pulsed mode (30 ns pulses, 10 Hz, 200 mJ/pulse) induces an efficient cell death. Thus, ca. 98% decrease in cell survival is obtained upon 5 min irradiation of cells that have been incubated for 48 h with 5.1 microM NiNc. The efficiency of the photoprocess is strongly influenced by the NiNc cell incubation time prior to irradiation. Photothermal sensitization with NiNc appears to open new perspectives for therapeutic applications, as suggested by preliminary in vivo studies with C57/BL6 mice bearing a subcutaneously implanted amelanotic melanoma. PMID- 10672536 TI - Skin photosensitization with topical hypericin in hairless mice. AB - Hypericin, a naturally occurring photosensitizer, exhibits interesting in vitro photobiological activities, which suggest that the compound is a potential antipsoriatic agent. In this study, the possibility of hypericin penetrating the skin in photo-active concentrations has been studied. Hypericin is incorporated in either emulsifying ointment supplemented with solketal (hypericin content: 0.05%) or in polyethylene glycol (PEG) ointment (hypericin content: 0.5%) and applied to the skin of hairless mice for 4 h. After removing excess ointment, the mice are then irradiated with different light doses using a 500 W halogen lamp. As a positive control, intraperitoneally (i.p.) administered hypericin (10 and 40 mg/kg) has also been tested. Erythema, desquamation and erosions are demonstrated in the mice treated with hypericin in emulsifying ointment with solketal using a light dose of at least 4.5 J/cm2. In general, these reactions correlate well with those of i.p. administered hypericin (40 mg/kg), indicating that hypericin incorporated in emulsifying ointment with solketal is well absorbed by the skin of the mice. However, for the i.p. administered hypericin (40 mg/kg), we could not evaluate phototoxic reactions in the group of animals that received a light dose of 108 J/cm2, as they all died 12-24 h after irradiation, indicating extreme photosensitization with systemic hypericin at higher light doses. On the contrary, there is no measurable skin photosensitivity induced by hypericin when incorporated in PEG ointment or when 10 mg/kg hypericin is i.p. administered. Our results show that hypericin incorporated in a suitable vehicle can be delivered to the skin in photo-active concentrations. Using a vehicle such as emulsifying ointment with solketal, it will be possible to explore the photo-activity of hypericin in the treatment of psoriasis and other skin diseases. PMID- 10672537 TI - Time-resolved detection of singlet oxygen luminescence in red-cell ghost suspensions: concerning a signal component that can be attributed to 1O2 luminescence from the inside of a native membrane. AB - For about ten years, it has been debated whether in principle it is possible to detect 1O2 located within the cell membrane by performing experiments with cell suspensions or even in tissue. In this paper we present our investigations on photosensitized red-cell ghost suspensions (RCGSs) and our strategy for the detection of luminescence of singlet oxygen (1O2) from the inside of the cell membrane. Using a very sensitive apparatus for time-resolved 1O2 detection, a very promising sensitizer and an adequate experimental strategy, a very small amount of the detected luminescence indeed can be attributed to 1O2 from the inside of the ghost membrane. PMID- 10672538 TI - Analysis of UV-B-induced DNA damage and its repair in heat-shocked skin cells. AB - The heat-shock response is a cellular defence mechanism against environmental stresses that is evolutionarily conserved from bacteria to man. Numerous reports demonstrate the beneficial effects of heat-shock protein induction on cell survival under toxic or oxidative stress, e.g., in cardiac and cerebral ischemia or prior to organ transplantation. However, there is little data on the effects of heat treatment on damage caused by UV irradiation. Applying three independent techniques, we have tested the influence of thermal pretreatment of skin cells (1 h, 43 degrees C) on the initial extent of UV-B-induced DNA damage and its subsequent repair. For cultured human epidermal keratinocytes and dermal fibroblasts we can show reduced levels of nucleotide-excision-repair-associated DNA strand incision in the comet assay. Moreover, immunostaining and flow cytometric quantitation of thymidine dimers immediately and one day after irradiation, respectively, reveal that the initial DNA damage is not (keratinocytes) or only moderately (fibroblasts) lower in heat-shocked cells as compared to untreated controls. However, excision repair of dimers is significantly attenuated during the first 24 h in both cell types. Furthermore, using a modified host-cell reactivation assay, we are able to demonstrate that repair of UV-B-damaged plasmid DNA is lower if the transfected cells are previously heat shocked. In summary, heat treatment (1 h, 43 degrees C) inducing heat-shock proteins reduces nucleotide excision repair of UV-B-mediated DNA lesions in fibroblasts and keratinocytes during the following 24 h. This is not necessarily caused by elevated heat-shock protein levels themselves. Possibly the direct thermal damage of repair enzymes is more severe than the potential protective effects of heat-shock proteins. PMID- 10672539 TI - In vivo detection and staging of epithelial dysplasias and malignancies based on the quantitative assessment of acetic acid-tissue interaction kinetics. AB - A novel approach to the problem of non-destructive detection and staging of tissue lesions is presented. The method relies on the in vivo quantitative assessment of the spatial and temporal alterations of light-scattering properties, induced in epithelial dysplasias and malignancies of the cervix and larynx, after topical application of acetic acid solution. Initial clinical trials show that the method is capable of detecting incipient lesions and that differences in the dysplasia and malignancy grade are clearly manifested in the measured temporal characteristics of the phenomenon. PMID- 10672540 TI - Effects of partial blood engorgement and pretest carbohydrate availability on the repellency of deet to Aedes albopictus. AB - The pretest availability of 10% sucrose solution and/or partial blood engorgement in Aedes albopictus Skuse significantly influenced mosquito attack rates and the time of repellent protection in laboratory bioassays. In 46 cm L x 38 cm W x 37 cm H cages used in USDA repellent tests, non-blood-fed and partially blood-fed mosquitoes attempted to bite at similar rates. In small cages (5 cm dia. x 4 cm H), holding individual females, mean mosquito attack rates were reduced when females were partially blood fed, compared with those not blood fed. The protection period from bites by Ae. albopictus using 25% ethanolic deet (N,N diethyl-3-methylbenzamide) increased significantly in small and USDA standard cages when females had pretest access to sucrose solution, compared with females starved for 12 h. Partial blood engorgement in mosquitoes affected repellent protection time in USDA standard test cages but not in small cages. PMID- 10672541 TI - A world checklist of genera, subgenera, and species of ticks (Acari: Ixodida) published from 1973-1997. AB - Researchers on ticks and tickborne diseases have been extremely fortunate in having at their fingertips the tick bibliographies produced by Harry Hoogstraal and his coworkers at the U.S. Naval facility at Cairo, Egypt, and by Mildred Doss and her colleagues at the U.S. Department of Agriculture laboratory at Beltsville, Maryland, USA. The Doss checklist of tick families, genera, species, and subspecies is now 25 years out of date, and the following checklist of one new genus, nine new subgenera, and 110 new species of Ixodida brings together the nomenclature on ticks produced during the last quarter century. PMID- 10672542 TI - Larval habitats of anopheline mosquitoes in the Upper Orinoco, Venezuela. AB - Survey of larval habitats of anopheline mosquitoes was conducted in Ocamo in the State of Amazonas, southern Venezuela. The sampled habitats belonged to three different hydrological types: lagoons (26 habitats), forest pools including flooded forest (16 habitats), and forest streams (4 habitats). Out of 46 habitats surveyed, 31 contained anopheline larvae. Six species were found: Anopheles darlingi, Anopheles triannulatus, Anopheles oswaldoi, Anopheles peryassui, Anopheles punctimacula, and Anopheles mediopunctatus. Anopheles triannulatus was the most abundant species. Significantly higher numbers of anopheline larvae, in general, and of An. triannulatus specifically were found in lagoons with submersed macrophytes and sparse emergent graminoids than in forest pools with detritus. PMID- 10672543 TI - Mosquito control and bacterial flora in water enriched with organic matter and treated with Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus formulations. AB - Three tests were conducted during July 17 to October 30, 1998 to study the impact of two mosquitocidal microbial agents on mosquito larvae and their contribution to bacterial flora in aquatic microcosms. Formulations of Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus strain 2362 (Bsph) were applied at various rates to outdoor tubs enriched with rabbit pellets and filled with irrigation water from a reservoir. Mosquito larvae were effectively controlled by all treatments; the magnitude of initial and persistent control depended on materials and dosages applied. Bacterial flora were assessed in the irrigation water as well as water in the enriched tubs before and after treatment with the microbial agents. The irrigation water contained 800-1000 total bacterial cells/ml. The populations of total bacteria and spore formers peaked on day 3 after enriching and filling the tubs, then declined progressively to the low levels at the end of the tests. After treatment, the numbers of Bti and Bsph spores in treated tubs prevailed at a dosage-dependent manner, their populations peaked at three hours after treatment, and declined progressively thereafter. The contribution of Bti and Bsph spores to the total bacterial flora was negligible but significant to the counts of spore-forming bacteria. The gram-negative bacteria made up more than 80% of the total bacterial flora during the test periods; and, of these, gram-negative rods constituted the greatest proportion, gradually increasing from the time of flooding to the end of the tests. Gram negative cocci also occurred in relatively great proportion, but showed a reverse trend as compared with the gram-negative rods, declining gradually from pretreatment to the end of the tests. Gram-positive rods (spore formers), including Bti and Bsph, occurred in low numbers in all the tests but increased slightly in treated tubs due to the addition of Bti and Bsph spores. Gram positive cocci occurred occasionally in some water samples. PMID- 10672544 TI - Field efficacy of fipronil 3G, lambda-cyhalothrin 10%CS, and sumithion 50EC against the dengue vector Aedes albopictus in discarded tires. AB - The efficacy of three insecticides, fipronil 3G, lambda-cyhalothrin 10%CS, and sumithion 50EC were evaluated against the dengue vector Aedes albopictus in discarded tires in Kuala Lumpur, Malaysia. The dosage given for each insecticide was 0.01 g of active ingredient/m2. Fipronil 3G was the most effective larvicide with a residual activity of up to two weeks, causing 88% mortality in Aedes albopictus. Lambda-cyhalothrin 10%CS was effective for one week causing 92% larval mortality and two weeks with 63% larval mortality. Sumithion 50EC had a residual efficacy of one week with 79% larval mortality. PMID- 10672545 TI - Reproductive biology of Lutzomyia shannoni (Dyar) (Diptera: Psychodidae) under experimental conditions. AB - Baseline biological growth data of Lutzomyia shannoni (Dyar) were compared under two experimental conditions within insulated styrofoam chests and in standard laboratory incubators. The developmental time from egg to adult was 67 and 52 days, respectively. Based on cohorts of 100 females in each experiment, horizontal life tables were constructed. The following predictive parameters were obtained under each of the two conditions: net rate of reproduction (23.5 and 18.0 females per cohort female), generation time (11.4 and 9.4 weeks), intrinsic rate of population increase (0.27 and 0.30), and finite rate of population increment (1.31 and 1.36). The reproductive value for each class age of the cohort females was calculated. The observed parameters were obtained under each experimental condition: net rate of reproduction (1.9 and 2.5 females per cohort female), generation time (11.7 and 9.6 weeks), intrinsic rate of population increase (0.05 and 0.09), and finite rate of population increment (1.06 and 1.10). Vertical life tables were elaborated and mortality was described for every generation in each cohort. In addition, for two successive generations, additive variance and heritability for fecundity were estimated. PMID- 10672546 TI - The fleas (Siphonaptera) of South Carolina with an assessment of their vectorial importance. AB - We document 25 species of fleas from South Carolina including new state records for two species, the ctenophthalmids Epitedia cavernicola and Rhadinopsylla orama. Host and other collection data, by county, are provided, including many new records amassed from 1992-1998 and some older records gleaned from the Clemson University Arthropod Collection. One flea species, the rhopalopsyllid Polygenis gwyni, is especially common and widespread in South Carolina, particularly on the cotton rat (Sigmodon hispidus) in coastal plain habitats. The largest number of flea species (8) from a single host species was recorded from the cotton mouse, Peromyscus gossypinus. Several flea species of potential medical or veterinary importance were recorded, some of which are potential vectors of pathogens, such as the agents of murine typhus, murine typhus-like disease, sylvatic epidemic typhus, cat scratch disease, and rodent bartonellosis. A host-flea list for South Carolina is included. PMID- 10672547 TI - Experimental studies of interactions between wild turkeys and black-legged ticks. AB - Wild turkeys are increasing in abundance and distribution in eastern North America, but their potential role as hosts for ticks, or as predators on ticks, is unknown. We performed two experiments, one to determine whether juvenile black legged ticks (Ixodes scapularis) feed successfully on turkeys, and the other to determine if turkeys depredate adult black-legged ticks in forest habitats. Of 550 larval ticks placed directly on 5 captive wild turkeys, none engorged and only 7 (1.3%) were recovered; the remainder apparently were consumed during preening. Of 165 nymphal ticks placed on the turkeys, 5 engorged and 8 unengorged ticks were collected; 152 (93.3%) were apparently consumed. Of 250 adult ticks introduced into forest enclosures exposed to turkey foraging, 89.5% were recaptured, which was not significantly different from the 92.2% recaptured in control enclosures from which turkeys were excluded. We conclude that wild turkeys are unlikely to host juvenile black-legged ticks in nature, and that turkey foraging is unlikely to reduce local density of adult ticks. PMID- 10672548 TI - Susceptibility of the malaria vector Anopheles culicifacies (Diptera: Culicidae) to DDT, dieldrin, malathion, and lambda-cyhalothrin. AB - The susceptibility of the malaria vector Anopheles culicifacies to DDT, dieldrin, malathion, and lambda-cyhalothrin was determined in Karnal, Yamunanagar, and Ambala districts of Haryana State, India. The vector population showed a high degree of resistance to DDT and dieldrin. The test mortality to DDT and dieldrin ranged from 25% to 28% and 18% to 20%, respectively, in Nadasahib of Ambala districts. The mortality of An. culicifacies to malathion ranged between 65% and 68%. All the tests with lambda-cyhalothrin resulted in 100% mortality of An. culicifacies. DDT and dieldrin resistance did not confer cross-resistance to lambda-cyhalothrin in An. culicifacies. PMID- 10672549 TI - Bacteria and mosquito abundance in microcosms enriched with organic matter and treated with a Bacillus thuringiensis subsp. israelensis formulation. AB - Bacteria and mosquito abundance were studied in outdoor tubs unenriched and enriched with 0.04% rabbit pellets during the winter, 1999. The irrigation water used to fill the tubs contained a total bacterial count of 1.15-1.35 x 10(3) cells/ml. Adding rabbit pellets for enrichment yielded a total bacterial count of 5.50-7.63 x 10(5) cells/g. Bacterial densities in unenriched water were significantly lower than in enriched tubs on every sampling day. When bacterial densities in both enriched and unenriched regimens reached peak populations on day 3 post-flooding, their numbers in enriched water were 25-fold higher than the unenriched water. Under cool weather conditions, mosquito oviposition activity was low and larval development was very slow. Egg raft counts and larval densities in enriched water were nevertheless higher than those in unenriched water. After reaching peak populations on day 3 post-flooding, the natural decline in bacterial densities in the top portion of enriched water without mosquito larvae was lower compared with that in water with larvae. In water with larval present, the decline of bacterial levels in top water was greater than in bottom water on day 7 post-flooding. VectoBac G, a granular formulation of Bacillus thuringiensis subsp. israelensis caused a reduction in larval numbers of 80, 93, 73% at the rate of 5.5 lb/ac and a reduction of 94, 93, 86% at the rate of 10.6 lb/ac on days, 1, 3, 7 posttreatment, respectively. After treatment, the reductions of bacterial densities in untreated tubs were greater than treated tubs. These results indicate that mosquito larvae play an important role in the decline of bacterial populations by their feeding activity. PMID- 10672550 TI - Effects of neem products containing azadirachtin on blood feeding, fecundity, and survivorship of Culex tarsalis and Culex quinquefasciatus (Diptera: Culicidae). AB - When late 3rd or early 4th-instar larvae of Culex tarsalis Coquillett and Culex quinquefasciatus Say mosquitoes were treated with sublethal dosages of neem insecticide until pupation, the blood-feeding activity of the resulting adults was essentially the same as that of untreated controls. In contrast, blood feeding activity was suppressed when newly emerged adults were fed continuously on 10 parts per million (ppm) or 50 ppm azadirachtin (AZ) in 10% sucrose solution for seven days. Fecundity was also reduced by the various neem treatments. When late 3rd or early 4th-instar larvae were treated with 0.010 ppm AZ to pupation, the resultant females had a lower rate of oviposition than did the untreated controls after a full blood meal. When late instar larvae were treated at 0.005 ppm and 0.010 ppm AZ, the resultant females produced smaller egg rafts after a full blood meal, as compared to the controls, but egg viability was not affected. In newly emerged adults feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose for seven days (before blood feeding), the oviposition rate, size of egg raft, and hatching rate of the eggs after a full blood meal were all reduced. When newly blood-fed adults were fed continuously on 10 ppm and 50 ppm AZ in 10% sucrose for five days, their oviposition rate was lower than controls in most cases, but the egg raft size and viability of eggs were not affected. In freshly blood-fed females topically treated with AZ with 1 or 5 micrograms/female, the oviposition rate and size of egg rafts were generally reduced. The females receiving topical treatment laid eggs and their hatching was not affected. The longevity of adult females feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose solution after emergence was reduced, whereas, the longevity of males was only affected at the higher concentration. PMID- 10672551 TI - Compatibility of Bacillus thuringiensis serovar israelensis and chemical insecticides for the control of Aedes mosquitoes. AB - The compatibility of the commercial aqueous Bacillus thuringiensis serovar israelensis (B.t.i.) formulation, Vectobac 12AS, with the chemical insecticides Actellic 50EC, Aqua Resigen, Resigen, and Fendona SC, for the simultaneous control of Aedes larvae and adults was studied by dispersing nine different formulations using a portable mist blower, in single story half-brick houses. The effectiveness of the treatment was evaluated by measuring the larval mortality, adult mortality, and droplet analysis at varying distances from the sprayer. Persistence of the larvicidal activity of the chemical insecticides and B.t.i was also determined by measuring the larval mortality in the test samples 7 days posttreatment. The sprayed particles in all the trials were 50-60 microns in size, indicating that the particles were those of mist spray. Test samples placed within 3 m from the sprayer gave the maximum larval and adult mortality. Chemical insecticides exhibited maximum larval mortality in the 1 h posttreatment test samples and it was comparable to the larvicidal activity of B.t.i. The larvicidal toxins of B.t.i were more stable and were able to affect sufficient larval mortality for 7 days posttreatment. The larvicidal activity of the mixtures, i.e., chemical insecticides with B.t.i, in the 1 h posttreatment test samples was not significantly different from the larvicidal activity of the chemical insecticides and it was comparable to the larvicidal activity of B.t.i alone. However, the larvicidal activity of the mixtures was significantly more than the chemical insecticides alone in the 7 days posttreatment test samples except for the Actellic 50EC and Vectobac 12AS mixture. In all the trials, with or without B.t.i, there was no significant difference in adult mortality, indicating that this B.t.i formulation, Vectobac 12AS, was not antagonistic to the adulticidal activity of the chemical insecticides. From this study, it can be concluded that chemical insecticides can be used effectively for both adult and larval control, but the chemical insecticides do not exhibit residual larvicidal activity. Hence, for an effective control of both Aedes larvae and adults, it is advisable to add B.t.i. to the chemical insecticides, as B.t.i is specifically larvicidal and is also able to effect extended residual larvicidal activity. PMID- 10672552 TI - Ticks of South Carolina (Acari: Ixodoidea). AB - County and host records are reported for 19 species of ticks from South Carolina: Amblyomma americanum, Amblyomma maculatum, Amblyomma tuberculatum, Aponomma latum, Boophilus annulatus, Boophilus microplus, Dermacentor albipictus, Dermacentor variablis, Haemaphysalis leporispalustris, Ixodes affinis, Ixodes brunneus, Ixodes cookei, Ixodes marxi, Ixodes minor, Ixodes scapularis, Ixodes texanus, Ixodes woodi, Rhipicephalus sanguineus, and Ornithodoros capensis. Ixodes woodi is recorded from South Carolina for the first time. Boophilus annulatus and Boophilus microplus probably no longer exist in South Carolina, and Aponomma latum is an exotic species that is not established in South Carolina. Brief notes follow each species. PMID- 10672553 TI - [Seasonal variations in stroke incidence in North-Eastern Poland]. AB - The authors analysed seasonal and monthly incidence of and mortality from ischaemic stroke (IS) in the region of Bialystok (North-Eastern Poland). 839 cases of IS (437 men and 402 women), aged 22 to 94 years, were hospitalised in the Department of Neurology, Medical Academy in Bialystok during the analysed period (1990-1997). Significant seasonality in IS incidence was observed in both sex groups, with a nadir in summer. The lowest occurrence of infarcts was observed in August in women, and in June in men. The peak month for IS, independently of sex, was January. 240 patients (28.6%) died of stroke or its complications during the analysed period. The occurrence of fatal IS followed also a clear seasonal pattern with peak in autumn. The authors attempt to explain this seasonal incidence and mortality pattern of IS in relation to variation in temperature, diet, way of life (holidays in summer), and biochemical blood changes, which occur in different seasons of the year. PMID- 10672554 TI - [Clinical significance of oxidation and acetylation genetic polymorphism in patients with Parkinson's disease]. AB - The relationship between genetically determined polymorphic oxidation and acetylation and susceptibility to some disease has aroused much interest. The aim of our study was to evaluate whether patients with Parkinson's disease differ from healthy persons in their ability to oxidize sparteine and acetylate sulfadimidine as model drugs. Oxidation and acetylation phenotypes were estimated in 50 patients with Parkinson's disease. The control group consisted of 160 healthy volunteers for comparison of oxidation phenotype and 60 healthy volunteers for comparison of acetylation phenotype. The phenotyping of oxidation revealed two distinct populations among 50 patients with Parkinson's disease: 47 persons (94%) were extensive metabolizers of sparteine and 3 persons (6%) were poor metabolizers. In 160 healthy persons, 146 persons (91.2%) were extensive metabolizers of sparteine and 14 persons (8.8%) were poor metabolizers. The difference between frequency distribution of PMs and EMs in healthy persons and in patients with Parkinson's disease was not statistically significant. The phenotyping of acetylation showed among 50 patients with Parkinson's disease 38 persons (76%) slow acetylators and 12 persons (24%) rapid acetylators. In 60 healthy volunteers the phenotype of slow acetylation was observed in 29 persons (48.3%) and rapid acetylation in 31 persons (51.7%). The prevalence of slow acetylators among patients with Parkinson's disease in comparison to healthy volunteers was statistically significant (chi 2 = 8.7677/p < 0.003). The results of our study may suggest that the slow acetylation phenotype is associated with increased risk of the development of Parkinson's disease. PMID- 10672555 TI - [Increased expression of aminopeptidase N on lymphocytes in multiple sclerosis]. AB - Aminopeptidase N is an ectoenzyme. Its expression on lymphocytes is the effect of cell activation. We studied APN expression on peripheral blood lymphocytes of multiple sclerosis (MS) patients and in the control group of patients with other neurological diseases (OND). We observed increased APN expression on lymphocytes of MS patients during acute exacerbation and in the course of chronic progressive MS compared to MS remission and OND groups. No such differences were found in CD11a molecule of LFA-1 integrin expression on lymphocytes. We suppose that APN expression on lymphocyte surface can be a sensitive marker of these cell activation in the course of MS. PMID- 10672556 TI - [Pineal cyst in MRI: clinical symptoms and analysis]. AB - Application of magnetic resonance imaging (MRI) in radiology allows to estimate and analyse pineal gland and pineal region pathology more precisely. We report 47 MRI brain studies of patients in whom pineal cyst was recognized as the only pathologic finding. MRI of the brain was performed because of clinical symptoms as headaches (32%), vertigo (26%) and altered behaviour (13%). Because of the common occurrence of pineal cyst in MRI brain imaging it seems to be important to decide whether these patients need neurosurgical intervention, especially if together with morphologic abnormality definite clinical symptoms exist. PMID- 10672557 TI - [Recording of vestibular myogenic evoked potentials in patients with disturbed inner function]. AB - In this study, we measured VEMP in patients with disturbed function of inner ear. We studied 4 patients with unilateral sensorineural deafness and normal excitability of vestibular organs; 6 persons with unilateral weakness of vestibular function and 10 healthy subjects. We found that in patients with unilateral weakness of vestibular function VEMPs had short latencies and diminished amplitudes whereas in patients with unilateral deafness VEMPs were unchanged. These data strongly suggested that VEMPs were generated by stimulation of vestibular and not cochlear part of inner ear-sacculus. Our data confirm that VEMPs are a highly objective technique, simple, secure and comfortable for patients to assess the vestibulo-spinal connections. PMID- 10672558 TI - [Results of postoperative treatment of patients after brain tumor removal based on the cerebral circulatory pressure index (CCPI) in comparison with ICP and CPP]. AB - The aim of the study was the assessment of prognostic value of CCPI in relation to the results of treatment of patients with increased intracranial pressure after brain tumour operative treatment. The results of treatment of 107 patients with intracerebral tumours according to neurological state at the introduction of treatment (GCS) and initial values of ICP, CPP and CCPI as well as final results of treatment (GOS) are analysed. A special normogram collecting all of these parameters was constructed, as well as the directions of treatment depending on the areas of MABP, ICP and CPP were established. The CCPI seems to be more a sensitive and earlier signals suggesting the urgency and the direction of treatment than ICP and especially CPP alone. There seems to be a great consistence of the patient state evaluation according to GCS, GOS and CCPI which is also a prognostic factor. All the patients with initial CCPI below 1.5 died while all the patients with CCPI over 3.0 in good and very good neurological state, and all the patients with initial CCPI between 1.5 and 3.0 had neurological deficits. PMID- 10672559 TI - [Analysis of own clinical material treated by conservative and aggressive control of intracranial hypertension]. AB - These problems arising during the treatment of increased intracranial pressure (ICP) were analysed on the basis of own clinical material with consideration of different therapeutic methods--pharmacological and aggressive. Statistical analysis (contingency tables) showed that the treatment consisting of dexamethason, mannitol and furosemid was the most efficient in the group of patient with increased ICP in the range 15-30 mmHg. In patients with ICP in the range 30-50 mmHg, where other aggressive methods of treatment had failed, the most efficient treatment was cranio-dural decompression. PMID- 10672560 TI - [The proliferative potential of human pituitary macroadenomas on the basis of morphometric analysis of nucleolus organizer regions]. AB - Cell kinetic information is an important adjuvant to histological typing and grading of human brain tumours. In the current study detection of the proliferating potential of adenomas was attempted using silver colloid technique for argyrophilic protein associated with nucleolar organizer regions (AgNORs). The expression of nucleolar organizer regions was assessed on paraffin sections in 28 pituitary macroadenomas which included all the main types. Nucleolar organiser regions have been quantified with computer-aided image analysis system in order to evaluate the mean area of AgNORs and their number relative to an internal reference, the number and areas of clusters of AgNORs and the area of the nucleus. In the pituitary macroadenomas there were 1.48 +/- 0.30 (mean +/- SEM) AgNORs per cell nucleus and corrected values were 1.66 +/- 0.27 (mean +/- SEM) AgNORs. The mean nucleus area was 867.18 +/- 203.91 pixels. The mean AgNORs size was 54.50 +/- 34.80 pixels. Correlation of nucleus area/AgNORs area was: r = 0.7085. On the number of the AgNORs it was possible to distinguish 3 subgroups of macroadenoma with different proliferative potential: low < = 1.367 (1.479); medium 1.367-1.733 (1.479-1.864) and high > 1.733 (1.864) AgNORs per cell nucleus and corrected number of AgNORs per cell nucleus respectively. Thus, this method provides useful information about the proliferative potential of human pituitary adenoma. PMID- 10672561 TI - [Dynamics of blood flow velocity in middle cerebral arteries in dyslexic persons]. AB - The aim of our study was to investigate the blood flow in middle cerebral arteries during the different forms of cognitive activity in dyslectic persons. Two group of subjects were tested. The first group consisted of 10 students with school difficulties, diagnosed neuropsychologically as having a particular form of dyslexia, i.e. dysgraphy or dysorthography. 6 of them were right lateralized and 4--left lateralized. The second contained 10 students without such problems. 7 of them were right lateralized and 3--left lateralized. We used four kinds of cognitive tasks, during which the blood flow velocity in MCA in left and right hemisphere was measured with the Transcranial Doppler method. The analysis of the results showed differences between the groups of blood flow velocity levels, without a difference in performance profile. The dysgraphic persons had significantly higher blood flow velocity in the right hemisphere compared to the reference group. The results suggest particularly important role of right hemisphere in dyslexic persons, which is consistent with results obtained by other authors. The analysis of lateralization showed that this factor influences significantly the blood flow velocity level--the left lateralized persons showed lower rise of blood flow velocity than the right lateralized, regardless of the kind of task and measured hemisphere. PMID- 10672562 TI - [Clinical autosomal dominating arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL)]. AB - CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is a diffuse disease of small arteries, predominating in the brain. It starts during mid-adulthood and is characterized by recurrent ischaemic events (transient or permanent), attacks of migraine with aura, severe mood disorders, subcortical dementia and at MRI white periventricular leukoencephalopathy. CADASIL is an autosomal dominant disease. The gene Notch3 on which the mutation was detected is located on chromosome 19. There is so far no specific treatment and death occurs after a mean of twenty years. PMID- 10672563 TI - [Radiological evaluation of pineal pathology and its regions]. AB - Contemporary neuroimaging procedures facilitate the differential diagnosis of pineal region pathology. They are of particular importance in malignant pineal tumours which may have non characteristic clinical presentation. Computed tomography and magnetic resonance imaging with intravenous infusion of paramagnetic contrast allow to determine tumour size and localization as well as its morphologic features and growth dynamics. The diagnosis obtained is a valid basis for treatment strategies. PMID- 10672564 TI - [Neuropsychological rehabilitation of patients with cognitive function disturbances in organic brain damage]. AB - A new neuropsychological method of rehabilitation of brain-injured patients with cognitive disorders is presented in the paper. The so called cognitive rehabilitation is based on computer programmes, prepared for typical syndromes of cognitive disorders. Research studies in this review refer to the effectiveness of that kind of therapy, indicating its confirmed values and some aspects to be explained yet. PMID- 10672565 TI - [Fixating techniques of craniocervical junction: selection criteria]. AB - Adequate choice of fixation technique at craniocervical junction depends on many factors: anatomical conditions at fusion site (e.g. anterior dislocations of the odontoid and rupture of the transverse ligament are contraindications for direct odontoid screw fixation. Sublaminar wiring and interlaminar clamps are useless in case of deficiency of posterior bony elements of C1 and C2 whether a result of laminectomy or destruction), bone quality (osteopenic bone is contraindication for screw techniques either transarticular or transpedicular). Enclosing of occipital bone into instrumentation may be difficult in wire and clamping techniques. In contrast screw techniques allow for easy grip the occipital bone. Screw techniques seem ideal in cases requiring enclosing of the occipital bone. The fusion rate at C1/C2 level seems independent of fixation techniques. When supplemented with external immobilization even biomechanically inferior wiring or interlaminar clamping provide nearly 100 rate of fusion. Screw techniques are technically demanding but they seem the method of choice when occipital bone is to be enclosed in instrumentation. PMID- 10672566 TI - [Sequence of clinical changes and evolution of MR images in a case of chronic SSPE]. AB - Slow course case of subacute sclerosing panencephalitis is reported. The onset was at the age of 16 years and death followed after 5 years. He was observed during nine successive hospitalizations and the authors describe the changing clinical symptomatology parallelly with successive EEG records and brain MR imaging evolution. Successive phases of the disease were associated with hyperintense changes in T2-weighted.images, and were situated initially in occipital lobes, and involved later on periventricular areas and finally the whole white matter of both cerebral hemispheres. Subcortical brain atrophy developed parallelly. PMID- 10672567 TI - [Adrenomyeloneuropathy: a form of X-linked adrenoleukodystrophy. Report of a family]. AB - A family with adrenoleucodystrophy linked to chromosome X (X-ALD) is reported. Three patients, one man (proband) and two female monozygotic twins, had adrenomyeloneuropathy (AMN) which is a form of the disease. The proband had characteristic changes in MRI with demyelination of the white matter in the cerebral hemispheres. Both women (one of them was proband's mother) has somewhat less severe AMN form, but in her twin sister the syndrome was much more intense. The clinical diagnosis of the disease was confirmed by biochemical investigations -determination of the level of very long chain fatty acids, ALDP protein and the activity of peroxysomal beta-oxidation. PMID- 10672568 TI - [Cervical pain as the only complaint in a patient with metastatic advanced lung carcinoma (case report)]. AB - Description of a case of metastatic advanced squamous cell lung carcinoma without any symptoms of the original focus. The only complaint of the patient almost throughout the course of the disease was cervical pain. Despite fast clinical course of the disease considerable adaptation of spinal cord to compression by metastatic focus in the vertebral column, may occur. Also numerous metastatic in bones or large foci in suprarenal glands may remain silent for a longer time of the disease. Examinations carried out: bons scinticsanning, CT of the abdominal cavity and MRI of cervical spine allowed to suspect that process with great probability, what was confirmed by autopsy. PMID- 10672569 TI - [Hemicrania continua: a case report]. AB - Hemicrania continua is a rare idiopathic headache of unknown etiology. The clinical course is characterized by usually unilateral, continuous headache. There are some clinical variants of pain character and other symptoms. Indomethacin (50-150 mg per day, rarely higher) leads to complete remission in all patients. Current diagnostic procedures (including neuroimaging) should be recommended in all cases to exclude organic cause of headache. The authors report a case of a 46-year old woman with 3 years history of drug resistant, unilateral headache. Complete remission after administration of indomethacin (75 mg TID) was achieved. Problems of diagnostic, clinical course and treatment of hemicrania continua are discussed. PMID- 10672570 TI - [Trans-sternal approach to the cervicothoracic junction]. AB - Cervicothoracic junction and upper thoracic spine down to T4 can be reached through anterior approach via sternotomy. Transsternal approach is the best route to gain access to lesions localized within vertebral bodies of the upper thoracic spine allowing for their resection, interbody fusion and replacement with bone cement. Consecutive modifications of transsternal approach evolved toward less extensive osteotomy from full median sternotomy, through manubriotomy with clavicle resection and partial lateral manubriotomy. Less extensive modifications provide limited lateral exposure of the spine and are more demanding technically. We present two cases of upper thoracic spine tumours operated on through full medial sternotomy. We believe that median sternotomy has several advantages over less extensive modifications: it is technically simple to perform for trained thoracic surgeon, safer as it provides better exposure of the mediastinum and thus sufficient control of great vessels including subclavian ones, gives better exposure of T3, T4 and even T5 vertebral bodies, allows perpendicular sight and attack to anterior surface of the upper thoracic spine and therefore good visualizing of the posterior longitudinal ligament and dura, do not destabilize shoulder girdle nor affect function of the upper limb. Additional caudal exposure of the thoracic spine as down as T5 can be obtained by dissecting a plane between the brachiocephalic vein, vena cava superior and ascending aorta. PMID- 10672571 TI - [Instability of atlanto-axial complex in a boy with isolated odontoid process]. AB - The authors present a case of a 14-years old boy, in whom, after a mild trauma, massive signs of cervical spinal cord injury appeared. Isolated odontoid process with instability of atlanto-axial complex proved to be the cause. In course of therapy with methylprednisolone and rehabilitation full recovery was achieved. The operative management consisted in posterior interlaminar stabilization C1-C2 with autologous bone transplant and titanium cable (Sof'wire). The etiology of this disturbance is discussed and operation method is presented. PMID- 10672572 TI - [Obituary to Professor Henryk Wisniewski]. PMID- 10672573 TI - [Report from the XVth day of child neurosurgery in Poznan on the topic of "Neonatal neurosurgery" (March 8, 1999)]. PMID- 10672574 TI - Hydrocephalus enters the new millennium: an overview. PMID- 10672575 TI - The CSF accumulator. AB - The ability of the central nervous system (CNS) to store and release fluid energy plays an important role in both health and disease. The stored fluid energy is the product of the fluid volume and pressure. How changes in CNS fluid (CSF, blood, or extracellular fluid) energy are distributed is determined by the compliance of the fluid containers and their arrangement. Hydrocephalus and related diseases not only interfere with the absorption of CSF but also interfere with the exchange of CSF in response to positional changes, cardiorespiratory and intraperitoneal energy changes. While shunts allow for the diversion of CSF when the intracranial energy exceeds the absorbing receptacle energy, they do not normalize the return of CSF to the intracranial compartment as needed with the intracranial blood volume or pressure decreases (the accumulator function of the CNS's CSF). A CSF shunt that has an artificial accumulator proximal to the valve can potentially restore the accumulator function towards normal and prevent some of the complications associated with CSF overdrainage. PMID- 10672576 TI - Brain biomechanics: mathematical modeling of hydrocephalus. AB - The considerable amount of literature on mathematical models of hydrocephalus and other brain abnormalities is critically reviewed. These models have various degrees of mathematical sophistication, and have influenced not only the diagnosis of hydrocephalus, but also its treatment with CSF shunts. The mathematical models are classified into two classes, pressure-volume models, and consolidation models. Advantages and disadvantages of both types are pointed out with a view to removing the confusion frequently generated by the technical aspects of the subject. The conclusion is reached that, while none of the current models are good enough to be of immediate use to the neurosurgeon, mathematical models are likely in the future to be a powerful tool for the understanding and the treatment of hydrocephalus, as well as other conditions related to brain biomechanics. The amount of mathematics has been kept to the absolute minimum, but it is cited and appended for those who would like to dig further into this fascinating area of research. PMID- 10672577 TI - Post-traumatic hydrocephalus. AB - The syndrome of post-traumatic hydrocephalus (PTH) has been recognized since Dandy's report in 1914. The incidence of symptomatic PTH ranges from 0.7%-29%. If CT criteria of ventriculomegaly are used the incidence has been reported to be from 30%-86%. Differences in diagnostic criteria and classification have contributed to the variation in reported incidence. The diagnosis of PTH is established using a combination of clinical, imaging and physiologic data. Symptomatic PTH is to be distinguished from post-traumatic ventriculomegaly resulting from atrophy. Symptomatic PTH patients are likely to improve when treated by shunting. Ventriculomegaly secondary to atrophy is less likely to respond to shunting. A series of traumatic brain injury patients at Wayne State University has been followed since 1989. The overall incidence of shunt placement in this group is 3.65%. Future studies of PTH should be aimed at refining diagnostic classification and criteria. Analysis of a large PTH population may then identify alterable risk factors in the early post-traumatic brain injury period. Minimizing these factors will help prevent subsequent PTH and obviate the need for shunting. PMID- 10672578 TI - Pathology of the hippocampus in experimental feline infantile hydrocephalus. AB - Hydrocephalus is responsible for many pediatric neurological deficits presumed to be caused by neocortical pathophysiology. Relatively little is known about the role of non-neocortical CNS structures in this condition. In the present work experimental infantile hydrocephalus produced by intracisternal kaolin injection was studied in a neonate kitten model. The hippocampal formation was processed for electron microscopy, and the neuropil of the CA3 region was examined in untreated, severely hydrocephalic and age-matched normal animals. Both macroscopically and microscopically the thickness of the hippocampus was not decreased. Hippocampal pyramidal neurons were found in varying stages of cytoplasmic densification, and dendritic and axonal processes exhibited hydropic cellular deterioration. The number of synaptic contacts was decreased. However, there was no indication of edematous extracellular space and the ependymal covering of the hippocampus was intact. The macroscopic structural integrity of the hippocampus, as well as the dendritic, axonal and synaptic alterations, suggest that the dark pyramidal neurons are the result of deafferentation, which may have profound effects on learning and memory. PMID- 10672579 TI - Pre-natal ventriculomegaly and hydrocephalus. AB - Ultrasonic imaging of the human fetal brain has allowed ventriculomegaly and hydrocephalus to be categorized. In this study 40 fetuses with ventriculomegaly and 21 with an Arnold-Chiari malformation and a myelomeningocele had ventriculomegaly that resolved, stabilised or progressed in utero. Within the progressive group were those with hydrocephalus, hydrocephalus being defined as expansion of the cerebral ventricular atria together with disproportionate increase in the head circumference. The prognosis for fetuses with resolving and stable ventriculomegaly was good, reflecting the fact that the ventricular dilatation in these cases was probably caused by delayed parenchymal and cerebrospinal fluid pathway development. Whereas the prognosis for progressive ventriculomegaly was generally poor, suggesting that the causes were likely to have been chromosomal, genetic, an infective agent or a catastrophic event which had an adverse effect on parenchymal development. The causes of hydrocephalus also adversely affected brain development but additional damage was caused by raised intracranial pressure. PMID- 10672580 TI - Cerebrospinal fluid flow through an implanted shunt. AB - Although the circulation of cerebrospinal fluid enjoys an apparent simplicity, its underlying basis is amazingly complex. Many factors influence how CSF flows through the human central nervous system. In the presence of hydrocephalus and subsequent shunt placement, this system is further complicated. The challenge before us then is to advance our understanding of the mechanisms that underlie CSF circulation, to place within that context the implications of shunt dependency, and to design a shunt system capable of simultaneous redirection of CSF and restoration of the CNS toward normal. Such a new shunt system would have to overcome not only those complications that result from hydrocephalus, but also those that result from the shunt's very placement. The purpose of this publication is 1. to provide an overview of the CSF circulatory system in the context of hydrocephalus and shunt dependency; 2. to explore resultant complications within the framework of those factors that influence CSF circulation within the human biological system as well as those that arise from the factors that influence CSF flow through a shunt; and 3. to propose in brief an alternative shunt valve designed to restore the accumulator function of the CNS toward normal. PMID- 10672581 TI - NMR spectroscopic evaluation of cerebral metabolism in hydrocephalus: a review. AB - Cerebral ischemia contributes to cerebral damage in hydrocephalus. Many studies have reported changes in cerebral blood flow and metabolism, supporting this hypothesis. Magnetic resonance spectroscopy (MRS) enables us to investigate cerebral metabolism in a non-invasive and longitudinal manner, thereby providing a promising way of evaluating pathophysiological changes in experimental and clinical hydrocephalus. In this review, the potential of 1H (proton) and 31P (phosphorus) MRS in the assessment of cerebral metabolism will be summarized, and a synopsis of in vitro and in vivo MRS studies in experimental and human hydrocephalus will be presented. Changes in high-energy phosphate metabolism, intracellular pH and lactate production in several MRS studies are presumed to reflect cerebral ischemia. In vivo information on neuronal damage, maturational delay and membrane phospholipid metabolism may also be derived from 1H and 31P MRS data. Technical, methodological and pathophysiological considerations, which are important for a correct interpretation and comparison of different MRS studies, will be discussed. Finally, we will draw some conclusions on the significance of these MRS findings and the applicability of MRS in the diagnosis and evaluation of clinical hydrocephalus. PMID- 10672582 TI - The initial treatment of hydrocephalus: an assessment of surgeons' preference between third ventriculostomy and shunt insertion. AB - Third ventriculostomy is an option for patients who have traditionally received a ventriculoperitoneal shunt. This study has been conducted to determine: 1. How common is third ventriculostomy as the initial treatment of hydrocephalus? 2. Does the frequency of third ventriculostomy vary among surgeons? 3. What factors influence surgeons' decision to choose third ventriculostomy? Surgeons completed a questionnaire addressing patient selection and technique factors. Nine case scenarios were reviewed by surgeons who were then asked to choose a ventriculoperitoneal shunt or a third ventriculostomy as the initial treatment. Forty-three responses were received. The proportion of new patients treated with third ventriculostomy varied widely (0%-100%, median 13%). This was not related to years in practice, type of training or presence of residents/fellows. Factors that increased the chance of a third ventriculostomy were triventricular hydrocephalus on CT/MR, isolated aqueduct stenosis, thin ballooned floor and tectal tumor. Factors that decreased the chance of a third ventriculostomy were dilated subarachnoid spaces, meningitis and head injury. The presence of myelomeningocele or age < 1 year were less likely to influence the choice of operation. Variation in the rate of third ventriculostomy as the first treatment for hydrocephalus is large. It is unlikely that this degree of variation can be explained by differences in patient populations. Further work to refine and disseminate the indications for third ventriculostomy is warranted. PMID- 10672583 TI - Hydrocephalus and the reproductive health of women: the medical implications of maternal shunt dependency in 70 women and 138 pregnancies. AB - An increasing number of women with cerebrospinal fluid shunts are surviving to child-bearing age, and are making independent decisions in regard to planning their families. As a result, a broad range of interdisciplinary health care professionals will require information about the management of these patients, especially during pregnancy and delivery. The purpose of this ongoing study is to gather comprehensive data from shunted women regarding their clinical history during pregnancy and within the six-month post-partum period. As part of this study, the following questions were addressed: 1. How does maternal shunt dependency influence the course of pregnancy and pregnancy outcomes? 2. What neurosurgical complications characterize this population of patients? 3. What complications of shunt dependency influence obstetrical management including pre natal testing and delivery? 4. What are the implications of shunt dependency with respect to general reproductive health concerns within this population? A total of 70 respondents, 18-41 years old and accounting for 138 pregnancies, completed a questionnaire providing information on maternal background, medical history, shunt performance during pregnancy, management of delivery, pregnancy outcomes, and unusual complications. One hundred three (103) pregnancies resulted in 105 live births including two surviving sets of twins; of these, 84 occurred in women with ventriculoperitoneal shunts (including both mothers who gave birth to live twins). Four women underwent therapeutic abortions, five delivered pre-term, one mother delivered a stillborn infant, and 16 experienced 32 miscarriages (including two ectopic pregnancies, and 33 fetal losses). Three women had seizures during pregnancy. Nine mothers reported an increase in headache activity during pregnancy. Twelve described abdominal pains during the course of pregnancy with anecdotal reports of increased frequency of painful episodes during the first and third trimesters. Twelve babies were diagnosed with congenital defects, including one pair of fraternal twins individually diagnosed with symmetric parietal foramina. Seven additional children were diagnosed with developmental disabilities including attention deficit disorder (ADD), attention deficit hyperactivity disorder (ADHD), pervasive developmental delay (PDD), and autism. Shunt malfunctions and revisions occurred seven times (four women) during pregnancy, and in 24 pregnancies (13 women) within six months of delivery. One malfunction and revision followed the miscarriage of twins at 12 gestational weeks. No acute malfunctions requiring immediate revision occurred during delivery, although two women reported severe headaches during labor. Transient signs of raised intracranial pressure occurred in 15 mothers over the course of 19 pregnancies which did not require surgical revision of the shunt following delivery or termination of pregnancy. No signs of shunt malfunction were identified in 100 of the pregnancies described in this series; 31 of these resulting in miscarriage and 69 resulting in live births. This study extends observations made previously to a larger population of shunt dependent mothers, and nearly doubles the amount of data available in our last publication. The results suggest that maternal shunt dependency carries a relatively high incidence of complications for some patients, but that proper management of these patients can lead to normal pregnancy and delivery. PMID- 10672584 TI - Progressive tissue injury in infantile hydrocephalus and prevention/reversal with shunt treatment. AB - Infantile hydrocephalus, despite shunt treatment, can leave children with a variety of persistent neurological deficits. A rat strain (H-Tx) with inherited fetal-onset hydrocephalus, is a natural model for the study of progressive tissue changes resulting from hydrocephalus and the effects of shunt placement. The cerebral cortex of rat pups has been studied at post-natal day 4 (P4), early stage hydrocephalus and equivalent to a third trimester human fetus, at P11, intermediate stage hydrocephalus and equivalent to a newborn human infant, and at P21 at advanced stage hydrocephalus. At P4, there is interstitial edema (increased water, sodium and chloride) and a non-reversible change in membrane lipids, particularly the phosphomonoesters. By P11, there are additional, non reversible, changes in intracellular potassium and energy metabolites (ATP and phosphocreatine). At P21, the cells are severely damaged and further intracellular changes include a decrease in N-acetylaspartate (NAA) and loss of amino acids and many organic osmolytes. The interstitial edema is approximately 75% reversed after shunt treatment. The loss of energy metabolites, NAA and osmolytes can be prevented by early shunt treatment at P4, but the subsequent potassium loss is not prevented. Shunt at P11 does not prevent loss of NAA or aspartate, but osmolytes are normalized. It is concluded that persistent tissue damage is initiated by changes in cell membrane components leading to a decrease in energy metabolism and loss of cell homeostasis. A more complete understanding of the mechanisms involved could lead to new approaches for therapy. PMID- 10672585 TI - Hydrodynamic principles in hydrocephalus: the engineer's perspective. AB - There have been significant improvements in the prognosis for patients suffering from hydrocephalus stemming from the introduction of the cerebrospinal fluid (CSF) shunt some 40 years ago. Currently, one of the major obstacles to effective shunt treatment is the mismatch between the physiology of the patient and the hydraulics of the shunt system. In order to maintain the proper relationship between CSF and cerebrovascular pressures, the implanted shunt needs to establish normal CSF outflow (absorption) and storage (compliance). Many of today's shunts establish a limited range of normal CSF outflow (absorption) and storage (compliance) once implanted, but a mismatch between CSF and cerebrovascular pressures may exist when the patient changes body position during daily activities. An uncoupling of these pressures creates mechanical strains within cerebral tissues, which are implicated in pathologies related to shunt malfunction. We suggest that re-establishment of normal CSF outflow resistance, which by definition is an indicator of both absorption and compliance, is a fundamental requirement for shunt treatment under most conditions. PMID- 10672586 TI - Fibroblast growth factor and hydrocephalus. AB - The immunohistochemical localization of basic fibroblast growth factor (bFGF) was studied in ventricular ependyma and choroid plexus of aged-matched normotensive and spontaneously hypertensive rats at different ages using a polyclonal antibody against bFGF. The bFGF-like immunoreactivity was observed in brain ependyma and choroid plexus of young and old normotensive rats. However, a progressive loss of immunoreactivity was observed with age in spontaneously hypertensive rats, that was associated with a progressive cerebroventricular dilation. These results show a new neuroendocrine anomaly to be added to the many others previously observed in this rat strain, when they develop hydrocephalus as they age. PMID- 10672587 TI - The living shunt: a tissue engineering approach in the treatment of hydrocephalus. AB - Tissue engineering is the use of cultured cells seeded into biodegradable polymers to create custom designed, living implantable devices. As a first approach to the use of this technique in the treatment of hydrocephalus, we have prepared chondrocyte-seeded polyglycolic acid (PGA) tubes coated with polylactic glycolic acid (PGLA), implanted initially with thin silastic stents removed four weeks after shunt insertion. The use of bovine xenograft cells in athymic (nude) rats resulted in more efficient seeding with chondrocytes, stiffer tube walls, and better patency. When implanted in 6-week-old rats made hydrocephalic by cisternal injection of kaolin at 4 weeks of age, six of eight 'living shunts' remained patent to radio-opaque contrast injection at two weeks after stent removal. At four weeks after stent removal, all four of the shunts had occluded at the ventricular end, three of the four apparently due to growth of the animal. We conclude that polymer type, cell type, and cell density will require considerable optimization, but a working tissue engineered shunt is feasible and may one day address some problems of interactions of living tissue and inert polymer. PMID- 10672588 TI - Magnetic resonance imaging study of extracellular fluid tracer movement in brains of immature rats with hydrocephalus. AB - Hydrocephalus is associated with brain compression and accumulation of neurotransmitter waste products in the brain and cerebrospinal fluid. We postulated that the extracellular compartment is compressed and specifically hypothesized that extracellular fluid tracer movement through brain would differ between control and hydrocephalic rats. Gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA) was injected into the cerebral cortex of 4-week-old rats, 7-11 days after induction of hydrocephalus by kaolin injection into the cisterna magna. The movement of this soluble paramagnetic compound was followed over successive timed intervals from 20 min to 180 min with T1-weighted magnetic resonance imaging. Non-hydrocephalic controls exhibited greater spread of the tracer and greater change in T1-weighted signal intensity in the ipsilateral cortex than hydrocephalic animals. Hydrocephalic animals exhibited preferential accumulation of tracer in edematous white matter. Gd-DTPA penetrated the lateral ventricles within 30 min in both control and hydrocephalic rats. The results suggest that there is a relative impairment of extracellular fluid movement through the cerebral cortex of young hydrocephalic rats. PMID- 10672589 TI - Genetic and embryological approaches to studies of neural tube defects: a critical review. NTD Collaborative Group. AB - Experimental embryological models have suggested that the morphology and quantity of neural tube defects may be governed by their position along the anteroposterior axis of the embryo. Inductive interactions and genetic regulation during axis development may play a role in the patterning of neural tube defects. A major challenge in the study of human neural tube defects is determining whether the spectrum of developmental neural tube anomalies found in individuals and their families mirror experimental models and are regulated by similar processes. We have found that the various neural tube defect phenotypes can be clustered according to their position along the anteroposterior axis. The findings correlate well to the pattern of early genes expression, inductive models of the embryonic axis, and mutant NTD animal models. We suggest that NTD should be studied by their location along the anteroposterior axis and that specific mutant genes may be identified by the observed pattern of NTD in an individual or a family. PMID- 10672590 TI - A brief review of the effects of chronic hydrocephalus on the gonadotropin releasing hormone system: implications for amenorrhea and precocious puberty. AB - Precocious puberty and amenorrhea have been associated with hydrocephalus, but the pathogenesis has not been determined. Approximately 22 cases of amenorrhea, and a few cases of precocious puberty, have been reported in hydrocephalic patients. Shunt treatment leads to initiation and maintenance of normal reproductive cycles in most cases. An underlying mechanism responsible for reproductive dysfunction may involve the role of gonadotrophin releasing hormone (GnRH). The exact pathway by which hydrocephalus disrupts the hypothalamic GnRH system is unknown. However, compressive forces, ischemia, and impairment of neurotransmitter feedback loops are likely candidates. PMID- 10672591 TI - Actualities in hydrocephalus classification and management possibilities. AB - Retrospective analysis in co-operative study of hydrocephalus at institutions of members of the Research Committee on Intractable Hydrocephalus sponsored by the Ministry of Health and Welfare of Japan was performed to determine the functional prognosis. For clinico-epidemiological study we classified non-tumoral hydrocephalus into eight types based on its etiology and the time of onset. Analysis of the 1,450 cases of hydrocephalus stored in the database obtained from the study was performed in order to find intractable factors in terms of factors related to patients and management. Analysis of the cases stored in the database revealed that the following types and conditions were found to be intractable factors: 1. Early fetal hydrocephalus. 2. Overt neonatal hydrocephalus. 3. Hydrocephalus associated with such severe brain malformations as hydranencephaly, holoprosencephaly and lissencephaly. 4. Hydrocephalus associated with severe brain damage. 5. Hydrocephalus associated with epilepsy. 6. Hydrocephalus shunted late after detection. 7. Hydrocephalus complicated by a shunting operation. It is impossible to determine prior to treatment whether or not a shunting operation is indicated for the patient with intractable factors, however, they may be a useful pre-operative indicator of prognosis. For the management of hydrocephalus, secondary intractable hydrocephalus may be preventable if we treat it appropriately before it becomes intractable. PMID- 10672593 TI - [The 1998 domestic state of development of cognitive enhancers]. AB - Recently, there is a great social problem that geriatric disorders, especially senile dementia, are growing rapidly with the increasing percentage of aged individuals in the total population. However, there is yet no drug that has reliable effects on senile dementia on the market. Therefore, society requires the development of new drugs that can support patients so that they can smoothly live their daily lives by themselves. In this study, we attempted to investigate the 1998 domestic state of development of cognitive enhancers by summarizing many publications and by gathering questionnaires from pharmaceutical, food, synthetic fiber and chemical manufacturing companies. There were 40 currently investigated cognitive enhancers in Japan as of the end of March, 1999 including 37 newly synthesized compounds and 3 new dosage forms or applications of additional effects. On the classification according to the mechanism of drug action, 12 of the investigated drugs are cholinomimetic agents, 12 are ameliorators of neuronal transmission, 1 is an intracellular mediative substance, 3 are neuropeptides, 2 are cerebral metabolic activators, 2 are cerebral circulation enhancers and 7 are neuronal cell protectors, and 1 is another type. For dementia of the Alzheimer's type, there are 1, 3, 10 and 2 drugs in prerecognition, phase late II, early II and I of clinical trials, respectively. For cerebrovascular dementia and cerebrovascular disease, there are 19 drugs being investigated. Seven of these compounds such as E2020 (donepedil HCl), DM-9384 (nefiracetam), TA-0910 (taltirelin), NS-3 (montirelin), TTC-909 (clinprost), DR-3305 (ebselen) and AVS (nicaraven) are at the prerecognition stage for marketing. It is important that effective cognitive enhancers will be supplied for clinical stage use as soon as possible. PMID- 10672592 TI - The effects of multiple shunt revisions on neuropsychological functioning and memory. AB - The focus of this study was to determine the effect of multiple shunt revisions on cognition and memory. The present study attempted to document a discrepancy in the functioning of children with hydrocephalus having numerous shunt revisions compared to those with only an initial shunt surgery. Researchers have found an increasing number of children with hydrocephalus requiring shunt revisions. In the current literature there are many conflicting views regarding the effects of hydrocephalus on cognition and memory. Many researchers report that properly treated hydrocephalus will not have a negative impact on cognitive functioning. Furthermore, researchers found that factors such as the total number of shunt revisions do not negatively impact global intellectual ability. Forty-six subjects between the ages of six and 16 years participated in the study. The subjects were recruited from the Department of Neurosurgery at an urban pediatric hospital. Specific inclusion and exclusion criteria were met. Independent variables for the study included shunt revisions, seizures, and Attention Deficit Hyperactivity Disorder (ADHD). Subject groupings were based on whether the subjects required multiple shunt revisions or single shunt placement and the presence or absence of seizures and ADHD. Dependent variables included the subject's performance on measures of cognition and memory. Measures of functioning included the Wechsler Intelligence Scale for Children--Third Edition and the Wide Range Assessment of Learning and Memory-Screener. The results of this study did not support the presence of cognitive or memory impairments as a result of multiple shunt revisions. Anecdotal findings noted that seizures were the only independent variable to significantly account for the observed variance in scores of cognition, specifically Full Scale IQ, Verbal Comprehension, and Perceptual Organization. PMID- 10672594 TI - [Molecular cell biology of presenilins]. AB - Alzheimer's disease (AD), the most common cause of dementia in the elderly, is a progressive neurodegenerative disorder characterized pathologically by the presence of senile plaques and neurofibrillary tangles in the brains of affected individuals. Senile plaques are composed of amyloid-beta peptides (A beta), a proteolytic derivative of the beta-amyloid precursor protein (beta APP). A subset of AD is inherited as an autosomal dominant trait (familial AD, FAD). Mutations in genes encoding beta APP, presenilin (PS) 1 and PS2 are known to cause FAD. Genetic mutations in all three genes that cosegregate with FAD increase the production of the most amyloidogenic species, A beta 42. Moreover, PS1-deficient neurons exhibit severe defects in the production of A beta, suggesting that PS1 plays an important role in gamma-cleavage that liberates the C terminus of A beta. The physiological role of PS is still unknown, but data from studies in C. elegans, Drosophila and PS1 knock out mice suggests that PS1 plays a crucial role in Notch signaling, and recently it was shown that PS1 is required for the proteolytic release of the intracellular domain of Notch following activation of Notch by its ligand. Further studies on PS-mediated intra- and jaxtamembranous proteolysis will lead to the understanding of the pathological mechanism of AD as well as of a novel mode of membrane protein processing. PMID- 10672595 TI - [Molecular pharmacology and physiology of nociceptin]. AB - The family of the G protein-coupled opioid receptors was recently extended by a novel member that did not bind any of the typical opioid receptor ligands. Identification of the orphan receptor in this way led to the advent of "reverse pharmacology" to identify the corresponding physiological ligands. Nociceptin, a heptadecapeptide, which was discovered as an endogenous ligand, first, attracted us by its reported nociceptive or anti-opioid actions. However, following studies revealed that this peptide has both nociceptive and antinociceptive actions under different conditions; e.g., administration routes or doses affect its actions. In our recent studies using a unique peripheral peripheral nociception test, nociceptin given locally at lower doses was found to produce nociception through substance P release from nociceptor endings, while at higher doses, it produced antinociceptive actions through an inhibition of phospholipase C activity stimulated by nociceptive substances. Such hypothetical mechanisms can be applied to the mechanisms of nociceptin-induced paradoxical actions in the central nervous system. The physiological role of nociceptin has recently been reported using nociceptin receptor knock-out mice. Following the report of a hearing problem in such mice, the nociceptin receptor was found to be involved in the development of morphine analgesic tolerance. In this review, more findings on the physiological roles of nociceptin or its receptor, such as pain control and memory-learning, are discussed on the basis of reports using nociceptin receptor knock-out mice. PMID- 10672596 TI - [Analysis of intracellular calcium dynamics in enterochromaffin cells of small intestine]. AB - Although serotonin (5-HT) release from enterochromaffin (EC) cells is considered to be regulated by multiple receptor-mediated mechanisms, little is known about the signal transduction. Here, we introduce the methods to isolate the mouse ileal crypts, which consist of various types of cells including EC cells, and to analyze the intracellular calcium dynamics. Ileal tissue was inserted with a plastic straw and the smooth muscle layers were peeled off. The mucosa were digested with collagenase and then suspended by moderate pipetting. Ileal crypts were separated by stepwise filtrations through 2 different nylon-meshes. The isolated crypt contained 0-3 EC cells as identified by immunostaining using anti 5-HT antibody followed by confocal microscopy. Isolated crypts were attached to a coverglass by an adhesive material (Cell-Tak) and loaded with fura-2/AM. Intracellular calcium dynamics in EC cells were obtained by digital video-imaging analysis of fura-2 fluorescence followed by the identification of EC cells with immunostaining of 5-HT granules. By these methods, it was suggested that norepinephrine elicited a transient elevation of intracellular calcium concentration in EC cells via alpha 2-adrenoceptors. These methods could be also useful to analyze the signal transduction system in intestinal endocrine cells that contain various intestinal hormones such as gastrin, cholecystokinin or secretin. PMID- 10672597 TI - [Usefulness of conditioned place preference (CPP) paradigm and its practical application]. AB - The conditioned place preference (CPP) paradigm is used to evaluate motivational properties such as rewarding or aversive effects of drugs. It was first introduced in the early 1980s to compensate for methodological and interpretive difficulties associated with the self-administration technique, the conventional method for assessing rewarding properties of drugs. The CPP paradigm has become the most frequently used method and its use has been reported more frequently than the self-administration paradigm. Although the CPP paradigm has mainly been performed in the rat, we have successfully established the CPP paradigm for the mouse. Thus, the CPP paradigm is now widely accepted as the behavioral approach to pave the way for the neural mechanisms of rewarding effect and for screening drugs for abuse liability. In this review, I focused on the significance, one's way of thinking, methodology, application and controversial points of the CPP paradigm. PMID- 10672598 TI - [Effects of nitrendipine on the development of hypertension and renal failure in Dahl salt-sensitive rats]. AB - The present study investigated the development of hypertension and the functional and morphological changes in the kidney in Dahl salt-sensitive (Dahl S) rats fed with a normal salt diet during aging. Furthermore, the effects of calcium channel antagonists nitrendipine and nicardipine on these changes were examined. The rats showed proteinuria from 6 weeks of age and gradually developed hypertension accompanied by the decrease in the glomerular filtration rate during aging. Glomerular screlosis and degeneration of the renal tubule were found by histological examinations at 17 weeks of age. Nitrendipine (20 mg/kg chow), given from 7 weeks of age for 10 weeks, inhibited the elevation in systolic blood pressure from 3 weeks after the dosing, whereas nicardipine (20 mg/kg chow) inhibited it only at 5 weeks after dosing. Both drugs decreased glomerular sclerosis, but did not affect the glomerular filtration rate, urine volume, urinary excretion of protein and N-acetyl-beta-D-glucosaminidase and serum concentrations of creatinine and urea nitrogen. These results demonstrated that Dahl S rats fed with a normal salt diet spontaneously developed the renal disorder in the early stage of hypertension and reinforce the validity of nitrendipine for the treatment of hypertensive patients with renal failure. PMID- 10672599 TI - [Correction of deformities of extremities I]. PMID- 10672600 TI - [Guided transosseous osteosynthesis. Modelling the size and shape of anatomical structures of the spine]. PMID- 10672601 TI - [Behavior of soft tissue structures in surgical leg lengthening]. AB - Experimental animal studies and clinical investigations show three processes going on during extremity lengthening with the Ilizarov-method. At first degenerative changes including cell necroses in the muscles, the nerves and the tendons [corrected] occur as well as denervation of muscle fibers resulting in neurogenous muscle atrophies. These alterations are followed by reperative and regenerative processes as well as the reinnervation of the denervated muscle fibers. Secondly histoneogenesis occur which leads to a high increase of tissue specific cells and a growth of the muscles, tendons and vessels. Therefore tensile-stress is an important factor of tissue growth. Thirdly adaptive processes proceed during leg lengthening. For example the high biosynthetic activity during tissue growth seems to indicate an increase of vasa vasorum. PMID- 10672602 TI - [Sonographic imaging of leg geometry]. AB - Posttraumatic malalignments are a frequent sequlae of IM nailing of lower extremity fractures. Conventional US has proven to be inferior to CT determinations of tibial or femural length and torsion. A new 3-D US method is presented that allows for accurate single step determination of lower extremity length and torsion without ionizing radiation. A regular US machine with a 5 Mhz linear probe is combined with an US localizer. Reference markers affixed to the lower extremity eliminate errors associated with patient position or motion. The 3-D US method was compared against CT (Ulm's method) in the measurement of torsion and length of the tibia and femur in 50 adults and 50 children. In both methods, the maximum difference of the intraindividual torsional angles and length measurements was 7 degrees and 7 mm. The maximum standard deviation for reproducibility in length measurement was 1.6 mm and 1.5 degrees for angular torsion. The new 3-D US technique was superior to CT in terms of reliability and reproducibility. Clinical advantages of the 3-D US technique include rapidity, independence from patient motion or positioning and the avoidance of ionizing radiation. Indications for 3-D torsional and length determinations include follow up evaluation of adult and pediatric tibial and femoral fractures, pediatric limb and gait evaluations, and osteotomy planning. PMID- 10672603 TI - [Correction of leg deformities. Definition, estimation and realignment of axis deviation and misalignment]. AB - The operative realignment of mechanical axis deviations (MAD) is necessary to prevent early joint degeneration and to reach normal load-bearing. Modern types of external fixation systems allow an alignment of the mechanical axis to exact degrees. Predominantly this are corrections of angular deformities. In some cases the analysis of the MAD does not show any angular deformity, but a parallel staggering of the mechanical axis lines of a bone. Such parallel staggering of the mechanical axis lines is defined as a translation deformity of the bone. In combined deformities with angulation and translation the centre of deformity can be established proximal or distal out of the limit of the bone. In translation deformities the realignment of the mechanical axis requires a parallel re staggering made by a translation-osteotomy or by a counter-angulated double osteotomy. In complex deformities with angulation and translation, the translation requires a separate corrective planning. By using external fixator systems to perform acute or progressive corrective osteotomies the position of the Axis of Correction of Angulation (ACA) in relation to the n-CORA of the deformity has to be considered. If ACA position is not conform with the n-CORA position relevant geometric effects in relation of length and translation occur. These geometric effects to corrective osteotomies can be calculated by using simple trigonometric formulas or graphical methods. Possibilities to compensate translation and length effects are shown by using unilateral external fixator systems. PMID- 10672604 TI - [Management of fibular hemimelia]. AB - Fibular hemimelia is a congenital longitudinal deficiency that represents a spectrum of deformities. The management of this condition is controversial and our treatment options are changing with developments in limb reconstruction techniques. For the severely affected child with a predicted limb length discrepancy greater than 25 cm at maturity and with a poor foot and ankle amputation is generally agreed to be the best option. For less severely affected limbs, particularly those with a predicted limb discrepancy of 10 cm or less and with a foot with 3 or more rays which can be made plantigrade, limb reconstruction is recommended. Controversy remains about the best way to manage children with an intermediate deformity. PMID- 10672605 TI - [Cogenital femoral defect. Indication, therapy and complication management]. AB - Congenital femoral deficiencies remain a surgical challenge. There is no common classification: both radiological and clinical methods are recommended. Depending on the severity of the deformity, reconstructive and lengthening techniques are performed, while accompanying deformations are taken into consideration. In addition, amputation or fusion techniques together with orthotic devices are used. A total of 35 patients (37 extremities) with congenital femoral deficiencies have been treated at the Orthopaedic Hospital Vienna-Speising, from 1982 to 1998. 24 extremities were treated with reconstructive and/or lengthening techniques. Results and complications are reported. PMID- 10672607 TI - [Leg lengthening and correction of deformity using the femoral Albizzia nail]. AB - The albizzia femoral nail allows for lengthenings up to 10 cm. It can be used in achondroplastic patients. Multiplane corrections can be addressed with a special nail (3D-Albizzia) if multiple osteotomies are performed (e.g. for proximal and distal femoral varus). A derotation and a flexion/extension osteotomy can also be associated. IM nailing is suitable for patient with previous external fixators. Surgical planning should be careful. The operation is currently performed percutaneously, with a 1 to 2 cm skin incision for nail insertion, and a 6 mm incision for distal femoral dome osteotomies. CPM machine is applied in recovery room and rehabilitation resumes on day 1, allowing more than 120 degrees of knee flexion by day 1. Ratcheting for gradual lengthening is begun on day 5 at a rate of 1 mm/day. Muscle stretching and strengthening are maintained for one year. In bilateral cases, a preoperative strengthening program is set. A 120 degrees knee range of motion can be maintained all during lengthening. Intramedullary lengthening allows for maintaining muscle and soft tissue suppleness which protects from hyperpressure over joints and from long-term muscle waisting. The Albizzia is currently a good tool once the surgeon get used to its physiotherapy aspect. Previously reported general anaeshtesia for getting the ratcheting is barely needed, when the technique has been done percutaneously and when the full range of knee motion is recovered on day 1 and maintained thereafter. PMID- 10672606 TI - [Fully implantable intramedullary distraction nail in shortening deformity and bone defects. Spectrum of indications]. AB - Since the first clinical experiences with the fully implantable programmable distraction nail nearly ten years ago, the system has been improved in Munich and meanwhile used in 26 patients. During the first 10 cases there has been highest interest in the reliability of the system, while in the following the expansion of indications was more important. At the thigh a good indication beside shortening is the combination of shortening and axis deviation, even if the center of deviation is located near to the knee joint in the supracondylar area. According to preoperative planing the deformity correction can be done acutely while the lengthening procedure follows postoperatively automatically at night time. If the stabilization with an intramedullary nail is possible, large bone defects can be treated by bone transport using this system also. The fully implantable intramedullary nail has proved its variable functions in cases of large bone defects combined with shortening of the femur. The system is able to perform the bone transport at first and the lengthening procedure automatically without any further operation thereafter. PMID- 10672608 TI - [External transpedicular fixation in the treatment of injuries of the spinal column and spinal cord]. AB - Injuries to the spine and the spinal cord are an important problem. In the present paper the application of external transpedicular fixation of the spinal column for surgical treatment of patients in the acute phase of spinal cord injuries is described. The results of treatment obtained in 54 patients in the acute phase are presented. Decompression of the spinal cord can be accomplished by way of a dorsal approach. In this way it is possible to remove the compressing factor in stages. Positive results were achieved in the late phase in 84.22% of patients with acute spinal and spinal cord injuries. PMID- 10672609 TI - Introduction: magnetic resonance imaging. PMID- 10672610 TI - From signal to image: magnetic resonance imaging physics for cardiac magnetic resonance. AB - Magnetic resonance imaging (MRI) is a powerful tool which enables the visualization of anatomy and the assessment of many physiological aspects of organ function. MRI and magnetic resonance angiography and magnetic resonance spectroscopy will play critical roles in cardiac applications during the next millennium. Thus, it is important to have a basic understanding of the most important physical processes in MR--the generation of nuclear magnetic resonance signals and their transformation into images. A conceptual description of these processes is the primary focus of this article. Also discussed are some additional signal manipulations specific to the needs of cardiac MRI, a field readily identified as the most significant in modern MRI technology development. PMID- 10672611 TI - Assessment of cardiovascular anatomy in patients with congenital heart disease by magnetic resonance imaging. AB - The following discussion addresses the assessment of cardiovascular anatomy in patients with congenital heart disease by magnetic resonance (MR). The focus of this review is on the techniques of performing the MR examination. In particular, individual pulse sequences are described and illustrated with their strengths and weaknesses. Imaging strategies using the described pulse sequences are proposed. The pulse sequences described are widely available on most MR scanners. Therefore, the proposed imaging strategies are clinically proven to be simple and effective ways to perform cardiac MR examination for the assessment of cardiovascular anatomy in patients with congenital heart disease. Functional imaging, such as flow analysis and ventricular function assessment, are discussed elsewhere in this issue. PMID- 10672612 TI - Three- and four-dimensional visualization of magnetic resonance imaging data sets in pediatric cardiology. AB - The purpose of medical imaging technology in pediatric cardiology is to provide clear representations of the underlying anatomy and physiology of the cardiovascular system--representations that are easily understood and that facilitate clinical decision making. However, standard projective and tomographic imaging methods often yield results that are intelligible only to imaging specialists. Three- and four-dimensional reconstructions from projective and tomographic data sets are an alternative form of image display. Often, these reconstructions are more readily comprehensible as representations of the reality apparent in the operating room or the pathology laboratory than are the original data sets. Furthermore, viewing of these reconstructions is much more time efficient than viewing hundreds of separate tomographic images. Magnetic resonance imaging inherently provides three-, four-, and even higher dimensional data, and magnetic resonance data sets are commonly used to generate volumetric reconstructions. This review will focus on the practical application of magnetic resonance imaging to yield three- and four-dimensional reconstructions of pediatric cardiovascular disorders. PMID- 10672613 TI - The range of normal values of cardiovascular structures in infants, children, and adolescents measured by magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) is a powerful diagnostic technique and research tool for assessment of congenital heart disease due to its ability to accurately assess anatomy, function, and flow in any orientation in the thorax. However, little data exist on normative reference values for cardiac structures, except in small study populations, and even fewer data exist for pediatric populations. In this review, MRI acquisition and analysis methods for assessment of aortic size, pulmonary artery size, and right and left ventricular function, volume, and mass are presented along with reference data obtained in pediatric populations by MRI. Where MRI data are not available, reference data obtained by echocardiography or angiography are included. PMID- 10672614 TI - Blood flow measurement by magnetic resonance imaging in congenital heart disease. AB - Investigation of blood flow in the heart and vessels may provide insight into the function of the cardiovascular system and aid patient management decisions. Phase velocity cine magnetic resonance imaging (PVC MRI) is a powerful and accurate noninvasive technique to quantitate and analyze blood flow. This article describes the principles, performance, and potential limitations of PVC MRI measurements. Clinical applications of PVC MRI are then reviewed with an emphasis on the assessment of congenital heart disease. PMID- 10672615 TI - Assessment of cardiac function by magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) is a unique and insightful tool for the assessment of physiology and function in congenital heart disease, in both the preoperative and postoperative state. MRI can accurately measure the volume and mass of unusual ventricular shapes, perform myocardial tissue and blood tagging, and can measure velocity and flow using phase-encoded velocity mapping. This has added new dimensions to research in pediatric cardiology. Newer techniques such as oxygen-sensitive MRI and echo-planar MRI promise further advances in the field. This article describes contemporary MRI studies of the physiology of complex congenital heart disease. PMID- 10672616 TI - Clinical applications of cardiac magnetic resonance imaging after repair of tetralogy of Fallot. AB - In the past 15 years, cardiovascular magnetic resonance (MR) has evolved into an imaging technique that provides adequate, and in part unique, information on residual problems in the follow-up of patients operated for tetralogy of Fallot. Spin-echo or gradient-echo cine magnetic resonance imaging allow detailed assessment of intracardiac and large vessel anatomy, which is particularly helpful in Fallot patients with residual abnormalities of right ventricular outflow and/or pulmonary artery. Multisection gradient-echo cine MRI can be used to obtain accurate measurements of biventricular size, ejection fraction, and wall mass. This allows serial follow-up of biventricular function. MR velocity mapping is the only imaging technique available that provides practical quantification of pulmonary regurgitation volume. MR velocity mapping can also be used to quantify right ventricular diastolic function in the presence of pulmonary regurgitation. PMID- 10672617 TI - Real-time magnetic resonance imaging: diagnostic and interventional applications. AB - The advent of ultra-fast imaging techniques has extended the utility of magnetic resonance imaging (MRI) from a static and purely diagnostic status to an imaging modality ideally suited for a number of therapeutic applications. These advances- along with the recent development and refinement of miniature intravascular imaging catheters and MRI-compatible guidewires, balloon catheters, and radiofrequency ablation catheters--have created an exciting forum of novel approaches for detecting and treating both acquired and congenital cardiovascular disease. This review covers the current state of the art in fast cardiovascular MRI, catheter-tracking techniques for MR fluoroscopy, and currently available interventional MRI systems. Early diagnostic and therapeutic applications, such as high-resolution intravascular and intracardiac imaging, balloon angioplasty, stent placement, and radiofrequency ablation techniques, are discussed and extended to several potential approaches specific to pediatric cardiac therapeutic catheterization. Lastly, safety aspects of MR-guided interventional procedures are presented. PMID- 10672618 TI - Microscopic and histochemical manifestations of hyaline cartilage dynamics. AB - Structure and function of hyaline cartilages has been the focus of many correlative studies for over a hundred years. Much of what is known regarding dynamics and function of cartilage constituents has been derived or inferred from biochemical and electron microscopic investigations. Here we show that in conjunction with ultrastructural, and high-magnification transmission light and polarization microscopy, the well-developed histochemical methods are indispensable for the analysis of cartilage dynamics. Microscopically demonstrable aspects of cartilage dynamics include, but are not limited to, formation of the intracellular liquid crystals, phase transitions of the extracellular matrix and tubular connections between chondrocytes. The role of the interchondrocytic liquid crystals is considered in terms of the tensegrity hypothesis and non-apoptotic cell death. Phase transitions of the extracellular matrix are discussed in terms of self-alignment of chondrons, matrix guidance pathways and cartilage growth in the absence of mitosis. The possible role of nonenzymatic glycation reactions in cartilage dynamics is also reviewed. PMID- 10672619 TI - Self-mutilation, masochism, and rigid character. PMID- 10672620 TI - Flesh made word: cutting back to the mother. PMID- 10672621 TI - Erotized transference and self-mutilation. PMID- 10672622 TI - Some reflections on self-mutilation. PMID- 10672623 TI - Further thoughts on "self-cutting": the intersubjective context of self experience and the vulnerability to self-loss. PMID- 10672624 TI - The lie: anorexia and the paternal metaphor. PMID- 10672625 TI - "A right-sided facial neuralgia": or fragmenting the history of the "Dora" manuscript. PMID- 10672626 TI - Abuse liability assessment of sibutramine, a novel weight control agent. AB - RATIONALE: Sibutramine (Meridia) is a serotonin and norepinephrine reuptake inhibitor marketed for weight control. Previous studies demonstrated low abuse potential for 20 and 30 mg sibutramine (doses near the therapeutic range); however, no data existed on supratherapeutic doses. This study, therefore, examined 25 and 75 mg sibutramine in humans compared to d-amphetamine (20 mg) as a positive control and placebo as a negative control. OBJECTIVES: The study examined the acute subjective, reinforcing, and physiological effects of sibutramine to assess its abuse liability. METHODS: Twelve polydrug abusers with no history of drug dependence participated in this double-blind, inpatient/outpatient study. Volunteers participated in four drug sessions, in which they completed subjective effects scales including the Profile of Mood States (POMS), Visual Analog Scales (VAS), and the Addiction Research Center Inventory (ARCI). The Multiple Choice Procedure (MCP) was used to evaluate reinforcing efficacy. RESULTS: Sibutramine 25 mg produced subjective effects that were indistinguishable from placebo. Sibutramine 75 mg produced significant unpleasant effects, such as Anxiety, Confusion, and decreased Vigor. On the MCP, volunteers chose to give up an average of $4.04 from their study pay rather than receive the higher dose of sibutramine again. In contrast, d-amphetamine 20 mg produced positive mood changes and was well liked. CONCLUSIONS: These data indicate sibutramine lacks amphetamine-type abuse liability when administered acutely. PMID- 10672627 TI - Activation of the mesocorticolimbic dopaminergic system by stimulation of muscarinic cholinergic receptors in the ventral tegmental area. AB - OBJECTIVE: To investigate the function of muscarinic receptors in the ventral tegmental area in vivo, the release of endogenous monoamines was simultaneously measured in the somatodendritic (ventral tegmental area) and terminal (frontal cortex and nucleus accumbens) regions of the mesocorticolimbic dopaminergic system in rats, using dual probe microdialysis. METHODS: Rats were implanted with dual microdialysis probes ipsilaterally into the ventral tegmental area (VTA) and nucleus accumbens (NAC) or frontal cortex (FC). RESULTS: Intrategmental infusion of the muscarinic agonist oxotremorine M (OXO M, 0.1 and 1 mM) increased extracellular levels of dopamine and serotonin, but not noradrenaline, in the VTA to a maximum of 200% over baseline in both urethane-anaesthetized and unanaesthetized rats. In freely moving animals, this effect was accompanied by strong motor agitation. Both VTA dopamine and serotonin levels dropped to 60% or less of baseline when the perfusion medium was replaced by a calcium-free medium containing OXO M. In the NAC and FC, a similar increase in extracellular dopamine, but not serotonin and noradrenaline, was observed during OXO M infusion in the VTA. The removal of calcium during OXO M infusion in the VTA did not cause a decrease in NAC dopamine levels. Activation of serotonin and dopamine release by OXO M in the VTA and FC was dramatically reduced or prevented by the co infusion of the muscarinic antagonist N-methylscopolamine (0.1 mM). CONCLUSION: These data demonstrate that VTA dopamine cells possess functional muscarinic receptors whose activation stimulates the release of dopamine in the VTA, NAC and FC. These results also suggest that muscarinic receptors may modulate the synaptic release of serotonin in the VTA. PMID- 10672628 TI - Sub-chronic inhibition of nitric-oxide synthesis modifies haloperidol-induced catalepsy and the number of NADPH-diaphorase neurons in mice. AB - RATIONALE: NG-nitro-L-arginine (L-NOARG), an inhibitor of nitric-oxide synthase (NOS), induces catalepsy in mice. This effect undergoes rapid tolerance, showing a significant decrease after 2 days of sub-chronic L-NOARG treatment. Nitric oxide (NO) has been shown to influence dopaminergic neurotransmission in the striatum. Neuroleptic drugs such as haloperidol, which block dopamine receptors, also cause catalepsy in rodents. OBJECTIVES: To investigate the effects of subchronic L-NOARG treatment in haloperidol-induced catalepsy and the number of NOS neurons in areas related to motor control. METHODS: Male albino Swiss mice were treated sub-chronically (twice a day for 4 days) with L-NOARG (40 mg/kg i.p.) or haloperidol (1 mg/kg i.p.). Catalepsy was evaluated at the beginning and the end of the treatments. Reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry was also employed to visualize NOS as an index of enzyme expression in mice brain regions related to motor control. RESULTS: L-NOARG sub-chronic administration produced tolerance of L-NOARG and of haloperidol-induced catalepsy. It also induced an increase in the number of NADPH d-positive cells in the dorsal part of the caudate and accumbens nuclei compared with haloperidol and in the pedunculopontine tegmental nucleus compared with saline. In contrast, there was a decrease in NADPH-d neuron number in the substantia nigra, pars compacta in both haloperidol-treated and L-NOARG-treated animals. CONCLUSIONS: The results give further support to the hypothesis that NO plays a role in motor behavior control and suggest that it may take part in the synaptic changes produced by antipsychotic treatment. PMID- 10672629 TI - A lack of tolerance to the anxiolytic effects of diazepam on the plus-maze: comparison of male and female rats. AB - RATIONALE: The demonstration of tolerance to the anxiolytic effects of benzodiazepines remains inconsistent. OBJECTIVES: The present study tested the hypothesis that intact and gonadectomized male and female rats might exhibit differential tolerance to the anxiolytic effects of diazepam (DZ). METHODS: Following acute (3 days) or chronic (3 weeks) DZ exposure, all animals were tested on the elevated plus-maze and immediately sacrificed for analysis of corticosterone, adrenocorticotropin hormone, estrogen and progesterone levels in serum. In experiment 2, following acute or chronic DZ exposure, animals were treated with a DZ challenge dose on the test day. RESULTS: In experiment 1, both acute and chronic DZ treatment similarly enhanced percentage open arm time and entries, regardless of the hormonal status of the animal. The results of experiment 2 showed that both acute and chronic DZ-treated animals exhibited a significantly higher percentage open arm time than control animals after the DZ challenge dose, and males and females did not differ in their responses to DZ exposure. CONCLUSIONS: The findings from these experiments suggest that tolerance to the anxiolytic effects of DZ did not develop in males or females, and that the hormonal status of the animal does not significantly alter the anxiolytic effects of DZ following either acute or chronic exposure. Following plus-maze exposure, females had significantly higher corticosterone levels than males and acute DZ treatment diminished this stress response. PMID- 10672630 TI - Caffeine withdrawal increases cerebral blood flow velocity and alters quantitative electroencephalography (EEG) activity. AB - RATIONALE: Cessation of daily caffeine consumption produces a withdrawal syndrome comprised of subjective symptoms and functional impairment. Few controlled studies have examined the physiological effects of caffeine withdrawal. OBJECTIVE: The present study examined the effect of caffeine withdrawal on cerebral blood flow velocity and quantitative EEG. METHODS: Ten volunteers reporting moderate caffeine intake (mean 333 mg/day) participated in this double blind study. Subjects completed several tests when maintaining their normal diet (baseline period) and during two 1-day periods during which they consumed caffeine-free diets and received capsules containing placebo (placebo test session) or caffeine (caffeine test session) in amounts equal to their baseline daily caffeine consumption. Blood flow velocity was determined for four arteries: right and left middle (MCA), and right and left anterior (ACA) cerebral arteries using pulsed transcranial Doppler sonography. EEG was recorded for 3 min from eight scalp sites while subjects sat, with eyes closed, in a sound-attenuated electronically shielded chamber. Subjective effects were assessed with questionnaires. RESULTS: Results showed an effect of the placebo (21-h withdrawal) condition compared to the caffeine condition. Placebo significantly increased the mean velocity, systolic velocity and diastolic velocity (cm/s) in all four cerebral arteries. In the MCA, the pulsatility index was significantly decreased following placebo. Placebo significantly increased EEG theta power. Placebo also produces subjective effect changes, including increases in heavy feelings in arms and legs and decreases in ability to concentrate. The caffeine and baseline conditions produced similar results on both the physiological and subjective measures. CONCLUSION: Cessation of daily caffeine consumption produced changes in cerebral blood flow velocity and quantitative EEG. These changes may be related to classic caffeine withdrawal symptoms of headache, drowsiness and decreased alertness. PMID- 10672631 TI - Effect of adjunctive paroxetine on serum levels and side-effects of tricyclic antidepressants in depressive inpatients. AB - RATIONALE: Previous studies showed that adjunctive paroxetine increases tricyclic antidepressant (TCA) serum levels by inhibiting cytochrome P4502D6. This effect has, however, been examined only in experimental studies using low doses of TCAs in healthy volunteers. OBJECTIVE: The present study investigated TCA serum level changes and side-effects after the addition of paroxetine in depressed patients treated with doses customarily used for inpatients. METHODS: 14 patients who had a moderate or severe depressive episode according to ICD-10 and who had not sufficiently responded (< or = 25% reduction of the Hamilton depression scale) to 3-week monotherapy with amitriptyline (n = 9) or imipramine (n = 5) with daily doses between 125 and 200 mg/day, received 20 mg/day paroxetine additionally under steady state conditions. RESULTS: After 2 weeks the serum levels of the metabolites nortriptyline (from 88 +/- 49 ng/ml to 176 +/- 57 ng/ml) and desipramine (from 152 +/- 78 ng/ml to 338 +/- 104 ng/ml) had risen to a significantly greater extent than those of the parent compounds amitriptyline (123 +/- 50 ng/ml to 195 +/- 128 ng/ml) and imipramine (from 75 +/- 36 ng/ml to 98 +/- 51 ng/ml). It is noteworthy that, with the exception of one case of incipient delirium, the combination therapy was well tolerated despite high TCA serum level rises. CONCLUSION: The higher increase of the metabolites as compared with the parent compounds can be explained by a paroxetine-induced inhibition of the liver enzyme cytochrome P4502D6, which catalyses the second step of the TCA metabolism, i.e. the hydroxylation of the metabolites. Blood levels should be meticulously monitored, if TCAs are combined with paroxetine. PMID- 10672632 TI - Effect of zopiclone and temazepam on sleep EEG parameters, psychomotor and memory functions in healthy elderly volunteers. AB - RATIONALE: The increased prevalence of sleep disturbance in old age is accompanied by a higher prescription rate of hypnotics, predominantly benzodiazepines in the elderly. In young volunteers zopiclone exerts a beneficial effect on sleep continuity without suppression of SWS and REM sleep; psychomotor performance and vigilance seemed to be less impaired than under classical benzoediazepines. OBJECTIVE: The present study investigates the effects of zopiclone on sleep EEG and cognitive performance in comparison to temazepam and placebo in the elderly population. METHODS: Single oral doses of zopiclone (7.5 mg), temazepam (20 mg) and placebo were administered in a randomized double blind, completely counterbalanced cross-over design to 12 healthy elderly men and women (65.9 +/- 3.6 years, range 60-70 years). On each of the 3 study nights a sleep EEG was registered from 10 p.m. to 6.30 a.m. and cognitive performance tests were applied at 8 p.m., 2 a.m. (when subjects were awake for 30 min), 7 a.m. and 9 a.m. RESULTS: After zopiclone treatment, sleep continuity had significantly improved and sleep stage 4 was increased compared to temazepam and placebo. In addition, both active substances significantly reduced REM density. Neither active compound substantially altered psychomotor and memory performance. CONCLUSIONS: Zopiclone and temazepam can be considered as effective hypnotics in elderly subjects when administered in that dosage. The superiority of zopiclone on sleep architecture may be related to a more specific action of zopiclone at the GABA-A benzodiazepine receptor complex. The suppression of REM density by both compounds and their subtle effects on cognition may reflect a GABAergic mediated reduction of cholinergic neuro-transmission. PMID- 10672633 TI - Effects of a low dose of alcohol on recollective experience of illusory memory. AB - RATIONALE: Memory illusions are currently a focus of memory research. Studies using the Deese/Roediger and McDermott paradigm have shown a differential pattern of illusory memories is associated with amnesia and ageing. The effects of pharmacological agents in this paradigm are not yet known. OBJECTIVE: Using this paradigm, the present study investigated the effects of a low dose of alcohol upon recollective experience of illusory memories. METHODS: A double-blind cross over design was used to compare the effects of alcohol (0.26-0.28 g.kg-1) with a matched placebo drink. RESULTS: High levels of false recognition were obtained across both treatments, replicating previous results. Although the small dose of alcohol employed did not produce gross changes in measures of false memory, it did modify the pattern of recollective experience in terms of remember and know responses. Specifically, it increased the level of remember responses for falsely recognised items (critical lures). CONCLUSION: These results are discussed in terms of ethanol's effects on false recognition of information which was not presented during the study episode. The effects of low dose alcohol on illusory memory are similar to the pattern found in ageing rather than that found in organic amnesia. PMID- 10672634 TI - Acamprosate, but not naltrexone, inhibits conditioned abstinence behaviour associated with repeated ethanol administration and exposure to a plus-maze. AB - RATIONALE: Drugs that reduce relapse in alcoholics are thought to inhibit either positive reinforcement for drinking (e.g. naltrexone) or negative reinforcement (e.g. acamprosate), and may reduce the impact of conditioned stimuli associated with previous alcohol use. We have developed a model for such conditioning by repeatedly pairing ethanol administration with plus-maze exposure. Substitution of saline for ethanol greatly increased stretched-attend postures and time in the central square, conditioned to the environment. OBJECTIVE: To test the hypothesis that if this behaviour indicates a negative affective state caused by the expectation of ethanol, it should be inhibited by drugs that reduce negative, but not positive, reinforcement. METHODS: The effects of naltrexone and acamprosate on alcohol-conditioned abstinence behaviour were compared. RESULTS: Acute administration of either drug alone produced no significant effects on plus-maze behaviour in naive mice. Naltrexone had no significant effect on the alcohol conditioned abstinence behaviour, but acamprosate reduced the incidence of stretched-attend postures. CONCLUSIONS: The experiments replicated previous findings for alcohol/environment conditioned behaviour, and demonstrated, as predicted, that this was decreased by acamprosate but not by naltrexone. Effects of acamprosate on conditioned negative reinforcement may be the cause of this effect, but more work is required to establish the usefulness of this model in evaluation of anti-relapse drugs. PMID- 10672635 TI - 5-HT3- and 5-HT2C-antagonist properties of cyamemazine: significance for its clinical anxiolytic activity. AB - RATIONALE: Cyamemazine is a neuroleptic compound which possesses anxiolytic properties in humans. On the other hand, 5-HT3- and 5-HT2C-receptors have been implicated in anxiety disorders and a previous binding study has shown that cyamemazine possesses high affinity for both serotonin receptor types. OBJECTIVE: The present study was undertaken to establish whether cyamemazine antagonizes 5 HT3- and/or 5-HT2C-mediated responses, and whether it compares with reference compounds. METHODS: Cyamemazine was tested for its ability to antagonize: (i) 5 HT3-dependent contraction of the isolated guinea-pig ileum and bradycardic responses in the rat and (ii) 5-HT2C-dependent phospholipase C (PLC) stimulation in rat brain membranes. RESULTS: In isolated guinea-pig ileum, cyamemazine potently and competitively antagonized 5-HT-dependent contractions (pA2 = 7.52 +/ 0.08; n = 5). In this test, cyamemazine was 5-7 times more potent (pIC50 = 6.75 +/- 0.13) than tropisetron (pIC50 = 6.02 +/- 0.04). In rats, cyamemazine i.v. antagonized 5-HT-dependent bradycardic responses with ID50% = 3.2 +/- 1.5 mg/kg (n = 4). Finally, in rat brain membranes cyamemazine antagonized 5-HT2C-dependent PLC stimulation with Ki = 424 nM (mianserin exhibits a Ki = 113 nM). CONCLUSIONS: Cyamemazine behaves as an antagonist at both 5-HT3- and 5-HT2C-receptors, which compares well with reference compounds. These 5-HT3- and 5-HT2C-antagonistic actions of cyamemazine can be involved, at least in part, in its beneficial therapeutic actions in anxiety disorders. PMID- 10672636 TI - Reduction of drug self-administration by an alternative non-drug reinforcer in rhesus monkeys: magnitude and temporal effects. AB - RATIONALE: Recent studies have shown that non-drug alternative reinforcers reduce drug self-administration. A goal of the present study was to explore factors such as magnitude of the alternative reinforcer and inter-session access to the alternative to identify conditions that lead to optimal reductions in drug intake. OBJECTIVES: To evaluate the effects of increasing the volume/delivery (v/d) of saccharin on oral phencyclidine (PCP) self-administration in rhesus monkeys given continuous access to PCP and saccharin during daily sessions using a behavioral economic analysis. The effects of availability of a saccharin solution during the inter-session period on session PCP consumption in drug experienced monkeys was also investigated. METHODS: Subjects had access to PCP (0.25 mg/ml) and either water or saccharin (0.03%) from two drinking spouts under concurrent and independent fixed-ratio (FR) schedules during daily 3-h sessions. The FR requirements for both available liquids were simultaneously increased (FR4 64). The v/d of saccharin or water was increased (from 0.3 ml to 1.2 ml), while the v/d of PCP remained constant (0.6 ml). In a second experiment, subjects had access to water or saccharin and water during the inter-session period (17.5 h) under an FR1 schedule. PCP and water were available during daily 3-h sessions under concurrent FR schedules. The FR for both liquids was increased (FR16-128). RESULTS: PCP intake was reduced at all FRs and magnitude conditions when saccharin (versus water) was concurrently available. Varying the v/d of saccharin only had a modest effect on the extent to which PCP intake was decreased at the higher FR values. Inter-session saccharin availability (versus water) reduced session PCP intake and the magnitude of this effect was also greater at the higher FR values. CONCLUSIONS: The magnitude of the saccharin delivery had an effect on PCP consumption at higher FRs, suggesting that economic factors such as high drug cost (FR) and low cost (responses/ml) of the alternative reinforcer (saccharin) interact to produce a maximum suppression of drug intake. Between session availability of saccharin also effectively reduced drug intake, and it had a greater effect on the maintenance levels of drug self-administration when the unit price of drug was high. PMID- 10672637 TI - 3'- and 4'-chloro-substituted analogs of benztropine: intravenous self administration and in vitro radioligand binding studies in rhesus monkeys. AB - RATIONALE: The reinforcing effects of many psychomotor stimulants have been related to increased dopaminergic neurotransmission. Drugs that block dopamine (DA) uptake have generally been found to function as positive reinforcers. Benztropine (BZT) and several of its halogenated analogs have previously been characterized as potent DA-uptake inhibitors with behavioral profiles that indicate diminished psychomotor stimulant effects relative to cocaine. OBJECTIVES: The present experiments were designed to examine, in rhesus monkeys, the reinforcing effects of the DA-uptake inhibitor BZT and two chloro-analogs 3' Cl-BZT and 4'-Cl-BZT, and to compare self-administration and binding profiles. METHODS: Four rhesus monkeys self-administered cocaine i.v. under a fixed-ratio 10 (FR10) schedule until stable responding was established. Saline, and various doses of cocaine, BZT, and the BZT analogs were then made available for self administration. Binding of these compounds to monoaminergic and cholinergic sites in monkey brain were determined using standard radioligand binding techniques. RESULTS: Self-administration was maintained by both 3'-Cl-BZT and 4'-Cl-BZT, but not by BZT. Results suggested that 3'-Cl-BZT and 4'-Cl-BZT were weak positive reinforcers. BZT and analogs bound DA transporters (DAT) with affinities higher than that of cocaine and had affinity for muscarinic binding sites. CONCLUSIONS: Surprisingly, high affinity at DATs was associated with weak or no reinforcing effects. The mechanism(s) that may underlie this dissociation between DAT actions and reinforcing effects remains to be established. These data support the proposal that a lead for the discovery of a pharmacotherapeutic agent for cocaine abuse may come from this group of compounds. PMID- 10672638 TI - A behavioural model to reveal place preference to delta 9-tetrahydrocannabinol in mice. AB - RATIONALE: The rewarding properties of delta 9-tetrahydrocannabinol (THC) are difficult to demonstrate in rodents using standard procedures. OBJECTIVE: To evaluate the motivational responses of THC in the place conditioning paradigm in mice after minimizing the dysphoric effects of the first drug exposure and/or the consequences of its pharmacokinetic properties. METHODS: Mice were conditioned to THC (1 or 5 mg/kg) using an unbiased procedure with an elevated number of pairings and long conditioning time. RESULTS: A place aversion was observed with 5 mg/kg THC using a standard protocol. Similar results were obtained when the CB 1 receptor antagonist SR 141716A (1 mg/kg) was administered immediately after each THC conditioning period. However, mice receiving a priming THC injection and conditioned 24 h later showed a place preference with 1 mg/kg THC and no effect with 5 mg/kg THC. CONCLUSION: THC produces a clear place preference in mice by using a long period of conditioning and avoiding the possible dysphoric consequences of the first drug exposure. PMID- 10672640 TI - The 1999 Crafoord Prize Lectures. The idea of information in biology. PMID- 10672639 TI - Experimental evidence that the aggressive effect of tryptophan depletion is mediated via the 5-HT1A receptor. PMID- 10672641 TI - The 1999 Crafoord Prize lectures. An evolutionist's perspective. PMID- 10672642 TI - The 1999 Crafoord Prize lectures. The Tithonus error in modern gerontology. AB - Tithonus asked Aurora for eternal life, when he meant eternal youth. Modern gerontological research makes the same mistake in its preoccupation with death, as if it were a programmed event in an organism's life history. Gerontology ought instead to investigate senescence, the decreasing effectiveness of mechanisms by which adult organisms avoid death or loss of fitness. Such studies should measure rates of decline in a diversity of adaptations and compare them within and between individuals and relate these rates and their correlations to genetic and environmental factors. The death of a studied organism must necessarily end its usefulness in providing valuable data. It is of little scientific significance. PMID- 10672643 TI - The 1999 Crafoord Prize lectures. Neo-Lamarckian experimentalism in America: origins and consequences. AB - The 1890s and the first decades of the twentieth century saw a vigorous debate about the mechanisms of evolutionary change. On one side, August Weismann defended the selectionist hypothesis; on the other, Herbert Spencer defended neo Lamarckian theory. Supporters of Spencer, notably the American paleontologist and evolutionary theorist Henry Fairfield Osborn, recognized that the questions raised by Weismann and Spencer could only be settled experimentally. They called for the application of experimental methods, and the establishment of a new institution for the purpose of confirming the inheritance of acquired characters. To a great extent, the experimental program championed by Osborn and others was implemented and, although it failed to reveal soft inheritance and was soon eclipsed by Mendelian and chromosomal genetics, it did make significant and lasting contributions to evolutionary biology. Thus the importance of methodological and institutional innovation and theoretical pluralism to the progress of science is illustrated and underscored. PMID- 10672644 TI - [Inseparable by definition and history, maxillofacial surgery and stomatology must build a program for the future together]. PMID- 10672645 TI - [What does the future hold for young maxillofacial surgeons?]. PMID- 10672646 TI - [Two or three things about the PMSI in stomatology and maxillofacial surgery]. AB - The Information Systems Medicalization Program (PMSI in French) was created in 1985 for Public Health Service Hospitalisation Structures. It appeared to be directly derived from the North-American Diagnosis Related Groups (DRGs) system. Since them, the PMSI has been progressively developed in private structures as well. The authors have had the opportunity to use the latest version of the computer program which was elaborated in order to share the patients into more than 500 different groups of pathology. These groups were called "Homogeneous Patients Groups" (GHM in French). To each group corresponds a "Synthetic Activity Index" number (ISA in French). It is supposed to be representative of the average cost of the management of each kind of patients, based on the diagnosis and the surgery possibly done. Several astonishing findings have been made. Some of them can be summarized as mentioned below: In maxillofacial Surgery, each group (GHM) seems in fact to be extremely inhomogeneous: for example, total parotidectomy with preservation of the facial nerve belongs to the same group as accessory salivary gland exeresis. Total skin graft is in the same group as free composite osseous flap with vascular anastomosis. Coding a surgical procedure leads often to reduce the ISA number in comparison with the same patient without surgery: "impacted third molar" gives 754 points without surgery but only 658 if surgery is performed. Carcinologic surgery is wholly grouped in the same category, even for rather short procedures as isolated partial glossectomy. This group corresponds to a great number of ISA points (6486) while bimaxillary surgery or free flap transfer gives less than 2500 points. In conclusion, the use of the PMSI to allocate financial means can be extremely dangerous for maxillofacial surgery units and consequently for the quality of the medical care in our Specialty. Further studies are obviously necessary to complete a critical analysis of the current system and to improve it. PMID- 10672647 TI - [The P.M.S.I. and maxillofacial surgery]. PMID- 10672648 TI - [Arteriovenous malformations in the jaws. The place of intravascular therapy. Apropos of 14 cases]. AB - PURPOSE: To analyze arteriovenous malformations (AVMs) of maxillo-mandibular arcades seen in our department since 1977, and to determine adequate treatment. MATERIAL AND METHODS: Fourteen AVMs were reviewed to determine their revelation, their semiology, the treatment applied and the results that could be obtained. All these lesions were true AV shunts involving bone with or without soft tissue extension. RESULTS: Eight patients belonged to the pediatric population. Six lesions were maxillary and 8 mandibular. All lesions were revealed during or after puberty by local swelling, pain, mass effect or bruit. Hemorrhage was the most frequent (71%) symptom. Teeth instability was detected in all these patients and was origin of the bleeding. The lesions were suspected clinically and confirmed radiologically. Angiography analyzed properly the architecture of the lesion (4 arteriovenous fistulas). Embolization was the clinical treatment in all patients: particles helped to stabilize the acute situations but failed to offer stable results, necessitating complementary embolizations and/or surgery (hemimandibulectomy in 2 patients). The use of acrylic glue (Histoacryl), injected percutaneously (5 patients), or transarterially in the AVM (4 patients)) provided cure in 6 of these lesions (43%) and long term stabilization for all the other AVMs. Teeth extraction could be performed thereafter in good conditions. Antibiotics and anti-inflammatory treatment helped to stabilize the evolutive risk of these lesions. CONCLUSION: Embolization is the therapy of choice in these lesions. Appropriate use of glue offers a high rate of cure and/or clinical stabilizations and avoids unneeded surgery. PMID- 10672649 TI - Antimicrobial effectiveness of 2% glutaraldehyde versus other disinfectants for hospital equipment, in an in vitro test based on germ-carriers with a high microbial contamination. AB - 2% glutaraldehyde is the reference disinfectant for hospital instruments. However, its high environmental toxicity makes desirable to search for alternatives. We compare the antimicrobial activity of 2% glutaraldehyde with 0.44% N-duopropenide (NDP), 0.66% NDP in 48 degrees alcoholic solution (NDP-alc), 0.13% glutaraldehyde-phenate, 1% or 3% persulphate (Virkon) and 0.1% or 0.5% chlorhexidine, using a model that mimics non-regular surface instruments contaminated with microbial strains (44 bacteria, 6 of which were Mycobacterium). The contaminated carrier is soaked in the disinfectant solution. After 5 or 20 minutes contact the disinfectant is neutralized. The overall results on all microorganisms in 20 minutes, show similar antibacterial activity for 2% glutaraldehyde and 0.66% NDP-alc, followed by 0.44% NDP and after by the two concentrations of Virkon and 0.5% chlorhexidine. The 0.13% glutaraldehyde-phenate and 0.1% chlorhexidine exhibited significantly less effect than any other disinfectant. 0.66% NDP-alc was faster antimicrobial activity than 2% glutaraldehyde, destroying totally the inoculum in 5 minutes. Activity on Mycobacterium showed great differences between 2% glutaraldehyde and the rest of products (> 5 log versus < 3 log reduction in 20 minutes), with an exception: NDP alc, with similar and faster activity (> 5 log in 5 minutes) than 2% glutaraldehyde. With human blood, the survival microorganisms increase 0.3 log (average) in all the disinfectants used. The aggressiveness on metallic devices was greater in Virkon than in the other disinfectants. We conclude that NDP (alone or in alcoholic solution) may be a good alternative to glutaraldehyde in hospital instruments disinfection. PMID- 10672650 TI - [Mandibular fractures in sports. Retrospective study of 48 cases]. AB - We present a retrospective study of 48 isolated mandibular fractures related to athletic activities. We studied patient age, sex, sport involved, mandibular location of the fracture and the therapeutic implication. The sex ratio was 4/1 and mean age 24 years. Rugby and cross-country biking were the more frequently involved sports (79%). We recall preventive measures. Miniplate osteosynthesis was used as often as possible in order to avoid intermaxillary fixation (IMF) (40%) or to limit the duration of IMF. This allowed early return to sport activities. PMID- 10672651 TI - [Sports and facial injuries. Feasibility and limitations of an epidemiologic study]. AB - Our aim was to conduct an epidemiological study in order to better assess the frequency of maxillo-facial sport injuries. Only those activities that are practised within a sport or game association are included in the study. A standard letter was sent to all French sport and game associations (Olympic or not) as well as to some major insurance companies (73 sport and game associations and 11 insurance companies). However the scope of the study rapidly proved limited since over the two-month period considered only 16 sport associations and one insurance company returned an answer. The exploitable results are presented and a brief overview of the recent relevant international studies is given. PMID- 10672652 TI - [An unusual fracture of the orbital floor]. AB - The various anatomopathologic forms of orbital fractures correspond to precise pathophysiologic mechanisms. We would like to detail an exceptional case of an orbital fracture in professional a diver. This case can be associated to barotraumatic accidents categories. After a brief review of the anatomy and the pathophysiological fractures of the orbit, we examine the different mechanisms which can be linked to this particular and unexpected pathology. PMID- 10672653 TI - [Pulsed dye laser: principles and indications]. PMID- 10672654 TI - [Eversion carotid endarterectomy: advantages and disadvantages]. AB - A comparative analysis of results of 70 operations fulfilled by the method of classical with autovein plasty of the internal carotid artery (ICA) and of 103 operations by the method of eversion carotid endarterectomy (CEAE) was made. The time of compression of the carotid artery during the eversion CEAE proved to be considerably less (22.5 +/- 6.5) min, than when using the "classical" method (32.5 +/- 5.3) min. In the nearest postoperative period no neurological complications of embologenic genesis or those associated with acute thrombosis of ICA after the eversion CEAE were noted while after the "classical" method they developed in 3 patients and 2 of them died. After the eversion CEAE 1 patients died of myocardial infarction. In remote periods after the "classical" CEAE the restenosis and reocclusions appeared in 6 patients, while after the eversion method--in 3 patients. CEAE fulfilled by the eversion technique is an effective operation reducing the amount of postoperative neurological complications as well as of late restenosis and reocclusions. It can be used without the applying of the internal bypass. PMID- 10672655 TI - [Clinico-morphological evaluation of physical methods of dissection and coagulation in stomach surgery (a preliminary report)]. AB - The article presents preliminary results of experimental investigations of clinico-morphological alterations in the gastric wall occurring during operations on the stomach performed with using high frequency electrosurgical apparatuses. The authors stress the expedience of using the apparatuses with the working frequency of electric current higher than 3 MHz for the abdominal surgery. PMID- 10672656 TI - [Motor-evacuation disorders of the digestive tract in the early period after stomach surgery. 3. Motor-evacuation disorders of the large intestine]. AB - The balloonographic method was used for studying the motor function of the sigmoid in 15 healthy volunteers, 15 patients with peptic ulcer before operation and in 25 patients at early terms after truncal vagotomy or resection of the stomach. During the first 2-3 days independent of the type of surgical intervention and of the degree of extra-organic denervation of the digestive tract, the motor activity of the sigmoid has the autonomic regimen of regulation, pathognomic structure of the motor cycle and is characterized by a sufficiently high contractile activity at the expense of frequent and regular generation of the migrating motor complexes. The restoration of the motor function is completed by the 5th-6th day. The states of normo-, hypo- and adynamia of the sigmoid were established on the basis of the contraction activity coefficient. The functional motor-evacuatory disorders of the intestine of the middle and grave degree are always accompanied by hypodynamia and transitory adynamia of the sigmoid. PMID- 10672657 TI - [Pancreatoduodenal resection in cancer of the head of the pancreas]. AB - The nearest results of surgical treatment of 375 patients with carcinoma of the pancreatic head and of 104 patients with cancer of other organs of the pancreatoduodenal area are described. Radical operations were made on 62 of them. Resectability was 16.5%, lethality--9.7%, postoperative complications were noted in 35.5% of the patients. PMID- 10672658 TI - [Cytoreductive surgery in the treatment of stage IV colorectal cancer]. AB - An analysis of immediate and long-term results of treatment of 34 patients with colorectal cancer of the IV stage has shown the expedience of cytoreductive operations in combination with postoperative chemotherapy and immuno-correction. Such complex treatment allows to increase the average survival to (27.8 +/- 4) months. PMID- 10672659 TI - [Ways of improving the results of surgical treatment of colonic obstruction of tumorous genesis]. AB - The work presents results of examinations and treatment of 522 patients with intestinal obstruction due to stenosing cancer of different parts of the colon. Operations were performed on 486 (93.1%) patients. It was established that the widening of indications to radical operative interventions on patients of all age groups independent of the level of localization of stenosing cancer of the colon was one of perspective ways for the improvement of results of surgical treatment. In patients with the acute obstruction of the left half of the colon it is expedient to complete radical operations making primary anastomoses. PMID- 10672660 TI - [Hemodynamic characteristics of the state of musculo-venous pump of the lower limbs and pelvis in patients with varicose veins]. AB - The method of phlebotonometry developed by the authors was used in an analysis of results of the state of musculo-venous pump of lower extremities and pelvis in 106 patients with varicose disease. The dynamics of phlebotonogram indices was studied during isolated functional exercises of the muscles of prelum abdominale, femur, leg and foot. In the first and second stages of the disease the function of both the single pumps and the whole pump system is disturbed but mildly and has no stage distinctions. Their pronounced impairment appears but in cases of a complicated course of the disease. In valvular incompetence of deep veins the musculo-venous pump looses its function: the greater the degree of valvular incompetence, the worse the functioning of this structure. PMID- 10672661 TI - [State and prospects of the development of war traumatology and orthopedics]. AB - The article deals with the achievements of the department of military traumatology and orthopedics in treatment of different traumas of the locomotor system, complications of these traumas and with different questions of treatment of orthopedic pathologies. It is shown that organisation of specialized orthopedic-traumatological centers in the system of military forces is necessary. PMID- 10672662 TI - [Injuries of the duodenum]. AB - During 10 years there were 94 patients aged 16-68 years with injuries of the duodenum. Their case histories were studied in order to establish the causes of complications and lethality. Among them there were 48 patients with knife wounds, 5 patients with gunshot wounds, closed trauma of the abdomen was diagnosed in 40 patients, 1 patient had a iatrogenic wound of the duodenum. There were 17 isolated injuries of the gut and 77 combined and multiple injuries. Patients with traumatic perforations in the duodenum made up 89.4%, 31% of them died. In 70 patients suture of the duodenum was put during operation, 10 patients had sutures and intubation of the duodenum, in 6 patients the injured gut was excluded, in 1 patient resection of the duodenum was made and primary anastomosis was formed. 28 patients (29.8%) died. Among the causes of the deaths were non-compensated blood loss resulting from hemorrhage from the vessels of the liver and other organs, combined and multiple injuries. Pyo-septic complications led to death of 12 patients, duodenal fistula was found in 4 patients. An analysis of the material has shown that most patients with traumas of the duodenum could be cured by suturing the incised or lacerated wound of the duodenum. Exclusion of the duodenum is thought to be a helpful addition to operation on the injured duodenum, especially on large wounds. Pancreatoduodenectomy is necessary but seldom in surgery of such traumas. PMID- 10672663 TI - [Fractures of the zygomatic-orbital complex: diagnosis and choice of the treatment method]. AB - The article presents an analysis of surgical treatment of 113 patients with fractures of zygomatic-orbital complex. Operations were fulfilled on 77 patients with using miniplates from titanium, on 25 men with application of the osteosuture, in 7 cases with a combination of the osteosuture and the miniplate, intrasinusal fixation was done on 4 patients. The problem of using computer tomography in a pilot survey of the patients is considered. A possibility to apply absorbable polymeric seam material (PDS-II) was shown in 24 observations. PMID- 10672664 TI - [Surgical treatment of patients with injuries of the spine of the thoracic and lumbar localization]. AB - Surgical treatment of 276 patients with uncomplicated (71.7%) and complicated (28.3%) injuries of the spine of the thoracic and lumbar localization was taken as the material for the article. The authors propose a rational strategy and tactics of operative treatment of patients with traumas of the spine and also they make an assessment of efficiency of some new surgical technologies. The volume and optimum terms of the decompressive-stabilizing operations on the spine are determined depending on the kind of the vertebral injury and the general state of the patient. The effectiveness of the present-day systems of stabilization of the spine (transpedicular constructions and plates for the frontal fixation of the spine) was demonstrated. PMID- 10672665 TI - [Characteristics of differential diagnosis and tactics of treatment of patients with fractures of the condyles of the femur and tibia]. AB - The author has shown that the traditional conservative and operative methods of treatment having a great number of unsatisfactory results and long period of treatment do not conform to the tasks which, in the ideal variant, must result in liquidation of consequences of the injury. Moreover, one of the main elements of these methods of treatment--continuous plaster immobilization--should be recognized as contraindicated. The program of treatment should include the use of special apparatuses allowing the reposition, firm lateral compression of the fractured fragments without an additional external immobilization, early movements in the injured joint which make optimum conditions for the consolidation of the fracture and simultaneous restoration of the joint function. PMID- 10672666 TI - [Hemorrhage in cerebral gliomas]. AB - A study of 48 clinical cases with supra- and subtentorial localization to reveal poor prognostic factors leading to intratumoral haemorrhage is offered. Haemorrhage most frequently occurs in patients older than 40 years and characterized by insult-like course. Poor prognostic factors of such condition are: astrocytic type of structure, giant size of tumor, cystic degeneration, high vascularity and partial removal of the tumour. PMID- 10672667 TI - [Injuries the esophagus]. AB - An experience with treatment of 30 patients with ruptures of the esophagus of different genesis is generalized. The differentiated approach to choice of the method of treatment and character of surgical procedures, wider indications to plastic interventions on the esophagus wound with its suturing layer-after-layer, wide disclosure of the mediastinum and its purposeful drainage, use of the modern detoxication means favoured the achievement of quite satisfactory results of treatment. Lethality was 6.7%. PMID- 10672668 TI - [Effects of ultraviolet irradiation of blood and sodium hypochlorite on aromatic amino acid metabolism in phlegmons of the maxillofacial region]. AB - The investigation was performed in 39 patients with the severe course of phlegmons of the maxillofacial area. The concentration of the phenyl-pyruvic and para-hydroxyphenyl-pyruvic acids in the diurnal urine was found to be elevated which suggested the disturbed functional state of the liver. Treatment including the ultraviolet irradiation of blood and sodium hypochlorite resulted in rapid normalization of the level of para-hydroxyphenyl-pyruvic acid and in lower concentration of phenyl-pyruvic acid. PMID- 10672669 TI - [Sphincter-valve gastroduodenal anastomosis in surgery of stomach ulcer]. AB - A method of gastroduodenoanastomosis has been developed which allows exclusion or considerable reduction of the pathological effect of the duodenogastric reflux upon the gastric stump mucosa after distal resection of the stomach in patients with gastric ulcer. Complex examinations of the patients after operations have shown that the developed by the authors sphincter-valvular gastroduodenoanastomosis facilitates the rhythmic-portion evacuation of the gastric contents and prevents the reflux of the duodenal contents into the gastric stump. PMID- 10672670 TI - [Does a sector resection of the breast cure nodal mastopathy?]. AB - Results of the clinico-morphological investigation of 265 patients with localized mastopathy who were submitted to sectorial resection showed that in the margins of the operative wound there were morphological signs of mastopathy in 252 (95.1%) patients. The results obtained confirm the opinion that structural alterations of the tissues known to be the essential feature of fibroadenomatosis can not be local, they are of diffuse character. So, the sectorial resection performed for localized mastopathy can not be radical and is of no therapeutic significance. The indication to surgical intervention must be determined not so much by the necessary treatment as by the real risk of hypo-diagnosis of breast cancer. So, there is no need to fulfil the sectorial resection for localized mastopathy. It is enough to make operation of less volume (excision biopsy). PMID- 10672671 TI - [Mediastinal lymphosarcoma complicated by recurrent hydropericardium]. PMID- 10672672 TI - [Effectiveness of the use of solcoseryl in intensive therapy of hemorrhagic shock]. PMID- 10672673 TI - [Two-stage rupture of the spleen]. PMID- 10672674 TI - [Aneurysm of the splenic artery in liver cirrhosis and portal hypertension]. PMID- 10672675 TI - [Perforation of the Meckel's diverticulum by a pine needle]. PMID- 10672676 TI - [Incarcerated right-sided direct inguinal hernia with incarceration of the Meckel's diverticulum and necrosis of its apex]. PMID- 10672677 TI - [Deontological views of surgeons-angiologists (data of a sociological questionnaire)]. PMID- 10672678 TI - [Development of surgery in the Sakha Republic (Iakutia) during the last 75 years]. PMID- 10672679 TI - [The results of the scientific activities of the organizations under the Ministry of Public Health of Ukraine in 1998]. AB - Data are submitted on the scientific potential of the system of the Ministry of Health of Ukraine, its constituent parts, some features of financing and volume of scientific research work done in 1998. Results are presented of scientific investigations in basic fields of medicine and priority lines of activity of medical science. The importance of introduction of scientific developments into the health care practice is emphasized. Prerequisites for further development of sectorial medical science under present-day social and economic conditions are analyzed. PMID- 10672680 TI - [The clinical course and histological characteristics of medullary hematopoiesis in acute leukemia in those who worked in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. AB - The course is analyzed of myeloblastic leukemia in twelve participants in the elimination of the aftermath of the Chernobyl Atomic Power Plant breakdown. The iliac trepanobiopsies bone-marrow hemopoiesis is studied. A gross depression of normal hemopoiesis has been revealed together with a fibrous transformation of the bone marrow, which fact suggests a profound damage to the hemopoietic micro surroundings, being a cause of ineffective hemopoiesis, of grave course of the illness, and of failure to respond to cytostatic therapy. PMID- 10672681 TI - [The immunohistochemical determination of p53 and of proliferating cell nuclear antigen in the epithelial nuclei of benign prostatic hyperplasia following the accident at the Chernobyl Atomic Electric Power Station]. AB - With the purpose of studying into the morphogenesis and proliferous activity of the prostatic epithelium under a long-term exposure to low doses of ionizing radiation there have been conducted comparative histological and immunohistochemical (expression of p53 and proliferous cellular nuclear antigen PCNA) investigations designed to study benign prostatic hyperplasia in patients living in those Ukraine territories affected by radionuclide contamination (group III), residents of Kiev (group II), and patients having been operated on before the Chernobyl accident, having constituted the control group I. It has been found out that the incidence of prostatic intraepithelial neoplasia (PIN), the level of nuclear expression of proteins p53 (in the PIN epithelium) and PCNA (in the epithelium of both benign prostatic hyperplasia and PIN) of patients in groups II and III are by far higher as compared with those in group I. The stroma of benign prostatic hyperplasia in patients of groups II and III was clearly different from that in the control group in that the former was characterized by apparent phenomena of hyalinosis, sclerosis, fibrosis, and extensive inflammatory infiltration, which changes can be explained by a long-term systematic exposure of prostatic tissue to low doses of ionizing radiation. PMID- 10672683 TI - [The sanogenic effects of reflex acupuncture therapy in regard to the pathogenesis of hypertension]. PMID- 10672682 TI - [The importance of endothelin-1 in regulating the functions of the cardiovascular system]. PMID- 10672684 TI - [Helicobacter pylori in the etiology and pathogenesis of chronic diseases of the digestive tract]. PMID- 10672685 TI - [The biological properties and clinical significance of interleukin-8]. PMID- 10672686 TI - [The clinical problems of autoimmune thyroiditis]. PMID- 10672687 TI - [The characteristics of left ventricular remodelling in hypertensive heart patients and the possibilities for its drug correction]. AB - In 85 patients with mild and moderately severe arterial hypertension presenting with signs of hypertensive heart, processes of remodelling of the left ventricle (LV) of the heart were studied using combined echocardiographic criteria. An evaluation was done of effects of different groups of antihypertensive drugs on the revealed changes in the LV structure and geometry. The patients were divided into four groups depending on the antihypertensive drug taken (atenolol, cozaar, norvask, enalapril). All groups patients were found to derive benefit from a 4 month monotherapy: against the background of normalization of arterial pressure (AP) structural-and-geometric indices for LV have gotten improved together with those for pulmonary gas exchange. The antagonist of receptors to angiotensin-II (cozaar) and the inhibitor of the angiotensin-converting enzyme (enalapril) were found to be the most efficacious drugs in the correction of the revealed disturbances. Improvement in LV geometry and regression of its hypertrophy were not directly related to AP normalization, witch fact indicates that there is no relationship between antihypertensive and cardioreparative activities of the drugs studied. PMID- 10672688 TI - [The mechanism of the development of a fat microembolism in myocardial infarct]. AB - It was found out with the aid of morphological methods that during the acute phase of myocardial infarction there develops fat embolism of the vascular bed of not only pulmonary circulation but also systemic circulation. One of the sources of fat embolism is the degenerated myocardium: during the process of its contraction fat drops move out of the cardiomyocytes and may penetrate into the capillary network. PMID- 10672690 TI - [The use of angiotensin-converting enzyme inhibitors in patients with chronic glomerulonephritis]. AB - Results are analyzed of a prolonged treatment with inhibitors of the angiotensin converting enzyme (captopril, Capoten, 100 to 150 mg daily, Renitec, 10 to 20 mg daily) in 53 patients with chronic glomerulonephritis. Of these, 23 patients presented with nephrotic syndrome in prehypertensive stage, 30 were in the stage of chronic renal insufficiency. The time-related course of proteinemia was studied as were indices for systemic hemodynamics, azotemia. Shown in the study was a significant effect of ACE inhibitors on proteinuria, indicators of systemic hypertension. A tendency toward decline in indices for creatinemia was noted. A concept is considered of multifactor effect of ACE inhibitors resulting in inhibition of progression of renal insufficiency. PMID- 10672689 TI - [Dynamic interphasic tensiometry of the blood in chronic hepatitis and liver cirrhosis]. AB - Studied with the aid of the computerized tensiometer MPT2-Lauda (Germany) was dynamic surface tension (ST) of blood serum in patients with chronic hepatitis (ChH), hepatic cirrhosis (HC), and in essentially healthy people, with n = 44, 32, and 68 respectively. ChH was shown to be accompanied by a significant increase in ST indices in the region of short (t = 0.01 sec) times of the surface life as well as by a decrease in that of medium (t = 1 sec) and long (t- >infinity) times. Rise in parameters of interphase tensiograms is a prognosis negative sign with respect to the development of HC. Correlations have been established of indices of blood ST to its content of protein, lipid, and inorganic substances and to morphological signs of affection of the liver as well. PMID- 10672691 TI - [The communicative form of sexual dysadaptation in neurosis in men]. AB - Forty-one couple with the communicative form of sexual dysadaptation including sexual aversion in which men suffered from different forms of neurosis (neurasthenia, hysteric neurosis, obsessional neurosis) were examined using the method of systems-structural analysis. The results of the studies made revealed causes and mechanisms of development of the considered form of sexual dysadaptation, with manifestations and consequences of the above dysadaptation related disorders of sexual health of the family described in some detail. A relation was found out between the form of neurosis in men and development of sexual dysadaptation. The necessity is substantiated to consider the revealed regularities in making a diagnosis or performing a correction of aversion in male subjects with neurosis. PMID- 10672692 TI - [Ultrasonic dopplerography in the diagnosis of vascular headache in children against a background of thiotriazoline treatment]. AB - Overall twenty-four children aged 10 to 15 years suffering from headache were examined. Doppler sonography technique employed in the study of vessels of the brain and spine reveals a high level of anomalous vessels of the brain and spine in the absence of manifest hemodynamic disturbances in the arterial bed of the vertebral artery and in apparent changes in the venous system. Those patients with headache having been exposed to thiotriasoline treatment demonstrated a positive dynamics of indices for the venous circulation as evidenced by dopplerography. PMID- 10672693 TI - [Computed tomography of the brain in suppurative meningoencephalitis]. AB - The studies made showed that one-tenth patient with purulent meningoencephalitis (PME) needs to be examined by axial computerized tomography (ACT) techniques. The following items are indications for ACT: apparent or progressive neurological symptomatology, protracted or relapsing course of PME, formation of the intracranial liquor hypertension, apparent changes in echoencephalography, and congestive disorders in the eyegrounds. Brain ACT permits finding a relevant policy of treating patients (surgery or conservative option) together with predicting the course and outcome of PME. PMID- 10672694 TI - [The possibilities for preventive diagnosis and nondrug treatment in drug allergy]. AB - The paper is dedicated to the matter of topical interest--that of diagnosing allergic reactions during the preclinical stage and of potentialities of clearing away adverse reactions to drugs without employment of medicinal agents. A characteristic feature of the paper is an original approach based on the authors' method. Reflected in the table are observations of the authors in respect of six entities versus analogous diagnosis without allergy and essentially healthy subjects--blood donors. The development of allergy is accompanied by a striking augmentation of CIC, which fact is to be considered in routine medical practice as a diagnostic criterion to be used together with the tests suggested by the authors. PMID- 10672695 TI - [An analysis of surgical treatment methods in femoral neck fractures based on a patent information search]. AB - Patent inventive discoveries in the USSR over the period 1949-1989 are analyzed together with the world patent discoveries, inventions, designs (Ukraine since 1989 included) over the period 1978-1998, which are divided into several groups different in principle, criteria of analysis are singled out, new notions of "cruel", dosage, dynamic compression are determined. By making use of principles of biological low-invasive metallo-osteosynthesis and new notions of "cruel", dosage, and dynamic compression optimal options were found for fixation of fractures of the femoral neck: low-traumatic nail set in the neck at an angle of 130 degrees fit with a small-sized diaphyseal put-in lamina and a local stimulation of union or spongiosa for the neck of the femur screw with an attachment preventing caput femoris rotation, and a springy element combined with a small-sized diaphyseal put-in lamina (that is, a modified system of the dynamic screw for the neck of the femur). PMID- 10672696 TI - [The conservative treatment of trophic soft-tissue lesions in patients with inveterate complicated spinal trauma]. AB - The paper presented is based on the analysis of 216 patients who had suffered a complicated spinal injury presenting with trophic lesions of soft tissues. The above pathology is found with very high frequency in patients with a severe degree of the spinal cord injury (groups A, B, C, and D according to Francel). Large soft tissue trophic lesions are long noted to be present being characterized by a high degree of microbial contamination. Our objective in this study is improvement of results of treatment of patients with a complicated spinal injury and the presence of trophic lesions of soft tissues with the aid of conservative methods of treatment such as electrostimulation and external laser therapy. A positive result of treatment was recordable in 30.9 percent of cases. PMID- 10672697 TI - [The effect of vertebral artery deformation by the osteophytes of uncovertebral areas on cerebral hemodynamics]. AB - Based on the analysis of 37 cases, an ultrasonic picture is described together with the character of a change in dopplerographic indicators in compression of the vertebral artery from osteophytes of the uncovertebral regions. In most cases (94.6%) the deformation is not accompanied by a hemodynamically significant compression (stenosing) of the artery and does not affect the indices for the intracranial bloodflow. The vertebral artery deformation from osteophytes affects parameters characterizing cerebral bloodflow when it is associated with other pathology. Movements in the cervical region of the spine do not increase compression of the vertebral artery from osteophytes of the uncovertebral region. Significance of the vertebral artery compression in the extracranial region can be adequately assessed mostly by indices for intracranial bloodflow. Thus, some other factors or their association that are most often not taken account of can play the leading part in the development of cerebral vascular insufficiency, such as occlusive lesions, deformations of arteries, anomalies of development, reflex vascular pathology etc. As a matter of fact, it is on extremely rare occasions that osteophytosis has a pronounced hemodynamically significant effect on the vertebral artery, it is not considered to be the only cause of cerebral circulatory disorders in people under 58-60 years of age. PMID- 10672698 TI - [The use of differential reflectometry in the EHF range as a new method in the diagnosis of ischemic heart disease]. AB - Described in the article is a new method of EHF differential reflectometry for ischemic heart disease (IHD), which relies on measurement of the anisotropy factor (AF) in sites of precardiac integumentum of patients in the millimeter range. A total of 30 patients were examined divided into six groups according to different functional classes. It is for the first time that anisotropy zones have been identified in IHD patients. The reflexogenic zone in the III intercostal space along the parasternal line was found to be of the most informative value. It is in this very zone that the largest AFmax values are achieved for myocardial infarction (MI) group exceeding the corresponding values for the rest of stenocardia groups by 20 to 100%. It follows from the foregoing that our method of MM-reflectometry permits diagnosing IHD together with differentiating the state of those patients presenting with angina pectoris and MI. PMID- 10672699 TI - [The medical rehabilitation characteristics of patients after therapeutic prophylactic operations in the early stages of rheumatoid arthritis]. AB - Overall 119 treatment and prophylactic operations were performed on those patients in early phases of rheumatoid arthritis who derived no benefit from a combined conservative treatment. With preliminary medial instability of the joint in 19 patients the surgical intervention involved some elements making for reinforcement of the capsular ligamentous apparatus. Good results of the above operations occurred in 78% of cases. Relapses of synovitis were encountered in eight percent of cases, which fact is related to the generalized advancing disease. Satisfactory results were obtained in those patients (14%) who were free from the origination of synovitis, which fact can be explained by lack of an efficient system of complex medical rehabilitation in the outpatient stage. A scheme is suggested of medical rehabilitation after treatment and prophylactic interventions for patients with early stages of rheumatoid lesions of the knee joint, that permits improving results of treatment in the above category of patients. PMID- 10672700 TI - [Intrathoracic compression of the respiratory tracts caused by broncho- and enterogenic mediastinal cysts in children]. AB - Over the period 1983-1999 examination was done together with a surgical treatment of ten children with bronchogenic (BG) and enterogenic (EG) cysts of the mediastinum, their ages ranging between 2 months to 13 years, having been kept under medical surveillance in the clinic for thoracoabdominal surgery of developmental anomalies in children. In 80 percent of patients intrathoracic compression of the respiratory tracts caused by the above cysts was diagnosed. 87.5 percent of patients had inflammatory bronchopulmonary problems, 12.5 percent had secondary tracheomalacia as complications of BG and EG cysts. BG and EG complications of the mediastinum affect the outcome of surgical treatment. PMID- 10672701 TI - [The comparative characteristics of the autonomic indices of the cardiovascular system in patients with combined gastroenterological diseases]. AB - As many as 32 patients with associated pathology of the digestive system were followed up. Studied in the above series was the cardiovascular system vegetal regulation-depended bodily adaptability. Those patients suffering from chronic gastroduodenitis with concurrent chronic cholecystitis and chronic pancreatitis were found out to have more manifest reactions of compensation and adaptation than patients presenting with duodenal ulcer with concomitant chronic cholecystitis and chronic pancreatitis. PMID- 10672702 TI - [The differential diagnostic laparoscopy of acute appendicitis and acute diseases of the internal genitalia in women]. AB - A total of 350 diagnostic laparoscopies were performed in the patients, we suspected, had acute appendicitis or acute surgical pathology of organs of the true pelvis. In patients with uncertain clinical signs laparoscopy yields a valuable information that allows the significant differential diagnosis to be carried out, with the optimal variant of the policy of treating patients to be selected in a timely fashion. PMID- 10672703 TI - [The characteristics of the course and treatment of caseous pneumonia in diabetic patients]. AB - Highlighted in the paper are particular features of the clinical course of caseous pneumonia in 18 patients with diabetes mellitus. An inter-aggravating character of the two illnesses is shown, especially in those patients presenting with a grave form of diabetes. The majority of patients have not derived much benefit from intensive chemotherapy against the background of alimentary therapy and insulinization. We consider it important to carry out surgical treatment in a timely fashion in a combined therapy of the above patients. PMID- 10672704 TI - [An analysis of the manifestations of the side effects of low-frequency ultrasonic therapy in pulmonary tuberculosis patients]. AB - It has been ascertained that employment of low-frequency ultrasound therapy in patients with pulmonary tuberculosis may provoke the appearance of such side effects as total and focal reactions. These side events are of temporary character, they are not considered to be contraindications to continuation of treatment with ultrasound. The focal reaction needs to be assessed x-ray carefully since its appearance can provoke aggravation of the specific process. PMID- 10672705 TI - [The efferent substitution correction of blood trace element disorder in patients with chronic kidney failure]. AB - Reasons are given for expediency of correction of dystraceelementemia in chronic renal insufficiency. Following exposure to the treatment with the multivitaminotraceelement drug three-we PLUS and alpha-erythropoietin eprex patients with terminal renal failure prolonged by chronic hemodialysis (n = 16) demonstrated a decrease in blood concentration of toxic trace elements and in dysbalance of essential trace elements, alleviation of clinical symptoms, increased tolerance to physical loads, improvement of hematological indices and quality of life. PMID- 10672706 TI - [The short-term effect of measured normobaric hypoxia on autonomic nervous system function in persons with hypo- and hyperacidic syndromes]. AB - The functional activity of the vegetative nervous system was studied before and after the exposure to graded normobaric hypoxia having been realized as breathing with an artificial hypoxic mixture (AHM) during 10 to 40 minutes in subjects presenting with hypo- and hyperacidic syndromes. A relationship was established between activation of the sympathetic or parasympathetic regions and aggravation of the antecedent history (bad habits). It is suggested that a short-term AHM breathing be used as a test for preservation/failure of mechanisms of adaptation. PMID- 10672707 TI - [The forensic psychiatric expertise of patients with epilepsy of traumatic origin and moderately pronounced mental disorders]. AB - A total of 17 patients were examined with epilepsy of traumatic genesis presenting with moderately severe mental disorder and unfavourable course of the illness. Criteria were defined more exactly of forensic psychiatric assessment of chronic mental disorders in this most challenging "borderline" (in the expert respect) group of patients. Established in these cases were conditions of use of juristical (psychological) criterion of the diminished responsibility formula: the presence of manifest changes in the personality with moderately severe ("borderline") deterioration of intellect, social and occupational disadaptation, predominance of psychopathological mechanisms in behaviour of patients including those during their committing illegal actions, rapidly progredient type of epilepsy course, and chronic alcoholism going in with the underlying condition. PMID- 10672708 TI - [The complex diagnosis and treatment of sexually transmitted infections and their complications]. AB - The article is dedicated to problems of diagnosis and treatment of those infections transmitted by sexual intercourse (STI) and their complications. A well-advised and feasible scheme of examining patients having applied for medical advice is suggested. Principal groups of drugs are described together with particular representatives of the above groups, the use of which will, we believe, help in delivering medical care to STI patients in a most efficient way. Levels of quality of delivering medical services are worked out depending on optimality of employment of different groups of drugs and their combinations in the treatment of STI and complications thereof. The data obtained during the studies made are to be used in the following circumstances: prescription by medical practitioners of particular schemes of STI treatment; control of quality of the health services delivery in medical settings dealing with those patients presenting with STI and complications thereof; planning of drug purchases by medical institutions providing profile medical care. PMID- 10672709 TI - [The effect of transcranial electroanalgesia on the neurophysiological indices in patients with cerebral venous dystonia]. AB - In the paper, results are presented of investigation into the effects of transcranial electroanalgesia (TCEA), a novel nonmedicamentous method of treatment, on certain neurophysiological parameters in patients with cerebral venous dystonia. TCEA is shown to be an efficient treatment option in the above group of patients. PMID- 10672710 TI - [The effect of chronic alcoholic intoxication on the development and course of diphtheritic polyneuropathy]. AB - The article reports in some detail on pathogenetic features of affections of the nervous system in those persons overindulging in alcohol. It is shown that the existing pathologies in the nervous system of the above patients get strikingly aggravated in diphtheria. Investigations designed to study the impact of chronic alcohol intoxication in 30 diphtheria patients overindulging in alcohol showed that there was a high case-fatality rate among persons falling into this category secondary to affections of the nervous system. Also common in this patient population were early developing grave lesions presenting as associations of the bulbar syndrome and peripheral disorders. PMID- 10672711 TI - [The effect of the initial level of immunity on the efficacy of antidiphtheria inoculations in children and adults]. AB - Based on results of examination in the passive hemagglutination test of 1440 subjects at different ages, several distinguishing features were revealed of formation of artificial active antidiphtheria immunity depending on the basic level of immunity. Single revaccination of those subjects presenting with the basic immunity of less than 0.03 IU/ml provides defence against diphtheria in only 33.3 percent of adults and 50 percent of children, in those subjects presenting with immunity between 0.03 to 0.99 IU/ml it is highly effective, in the immunity 1 IU/ml and beyond the effect of further immunization is very low since 25 to 33.3 percent of subjects demonstrate enhancement of immunity, whereas 16.7 to 25 percent show lowering of it. The analysis of the immunological structure of the population shows that 45 to 60 percent of adults in different age groups need to be exposed to single revaccination, 14 to 37 present will find it insufficient, 3 to 36 percent redundant. We suggest that revaccination against diphtheria be conducted under control of the level of antitoxic immunity. PMID- 10672712 TI - [The systems analysis of protective-adaptive reactions in immunopathology]. AB - In the systems approach to the study into the closed control circuits (CCC) of the self-regulation apparatus two combinations have been established of functioning (variable) quantities. A decrease in somatotropin and hypoglycemia and correspondingly an increment in somatostatin and insulin determine the redundant measure of defence against injury, the converse values in the analyzed parameters determine autoimmune processes. Alternation of the above parameters accompanied by exacerbation-remission and remission-exacerbation is directed to securing an adaptive final effect--recovery. Apparent structural-and-functional disturbances in certain regulators of any CCC are accompanied by persisting pathological conditions listed for each heading. PMID- 10672713 TI - [Hereditary predisposition in the formation of allergic diseases in children]. AB - Results are submitted of investigations designed to gain insights into the part hereditary aggravation (HA) in respect of atopy plays in the origination of allergic disorders (AD) in early childhood and to study objective markers of atopy and bronchial asthma (BA). Close correlation (P < 0.01) was established between HA and AD development, increase in the total IgE level and skin scarification tests. In BA, manifest bronchial hyperreactivity is accompanied by rise in total IgE, which observation can be related to their associated inheritance. PMID- 10672714 TI - [The use of Doppler echography in the diagnosis of diffuse toxic goiter]. AB - Echodopplerography demonstrated a significant increase in the velocities of bloodflow and indices for the thyroid gland in diffuse toxic goiter. The presence of the increased velocity indexes and coefficients during remission permits predicting recurrent course of the affliction. PMID- 10672715 TI - [Ozone hemo- and antioxidant therapy and endogenous intoxication in gestosis]. AB - The content was studied of the endogenous intoxication syndrome markers (medium size molecules) in red cells, blood plasma and urine of patients with grave gestoses in view of the fact that underestimation of the syndrome considered tends to significantly narrow the broadness of pathogenetic notions about the given pathology and to negatively influence the effectiveness of intensive therapy of patients. Besides, the problem gets more complicated because of pathogenetic specificities of severe gestoses (the use of the well-known methods of detoxication is made difficult in the patients). Ozone hemotherapy as an efferent method of detoxication and employment of antioxidant in the complex of intensive therapy of patients with severe gestoses make for lowering the degree of catabolic disturbances by decreasing the level of medium-size molecules in erythrocytes, blood plasma and urine. PMID- 10672716 TI - [The mechanisms for the loss of correction in the surgical treatment of severe forms of scoliosis]. AB - Of the 2719 patients presenting with grave forms of scoliosis who had undergone 4377 correction operations, 85 patients demonstrated serious complications in respect of loss of correction, with six of these having developed myelopathy in the long-term follow-up. Causes of the above problem included failure of internal fixation with distractors through the breakage of the construction or supporting structures of vertebrae and disturbances in external immobilization. As to its mechanism, loss of correction occurred because of transplant incapsulation or rearrangement of bone and plastic material, with the Loozer's zones, pseudarthroses, or defects in spondylodesis being developed. PMID- 10672717 TI - [The role of determining specific pregnancy proteins in the diagnosis of progressing tubal pregnancy]. AB - The importance was studied of measurement of certain specific pregnancy proteins (b1-glycoproteid, SP-1, PAPP-A, PP-5) for the diagnosis of advancing fallopian pregnancy. The data secured suggest a statistically significant decrease in SP-1 and PAPP-A concentration in tubal implantation of a fetal egg as compared to uterine pregnancy. Changes in blood serum concentration of PP-5 in women with different variants of the fetal egg implantation were statistically insignificant, which fact suggests that measurement of the quantitative indices of the pregnant women blood serum SP-1 and PAPP-A may be regarded as an investigational method of sufficient information value to be used in detection and differential diagnosis of progressive pregnancy in a timely fashion. PMID- 10672718 TI - [The clinico-hemodynamic effects of losartan in treating patients with chronic heart failure]. AB - Clinical and hemodynamic effects were studied of the blockader of AII receptors lozartan in patients with chronic cardiac insufficiency (ChCI). Inclusion of losartan into the conventional therapy of ChCI is accompanied by a lowering of the functional class in patients and improvement of haemodynamic maintenance of the physical exercise performance as evidenced by veloergometry. An 8-week's course of lozartan therapy leads also to a decrease in the anterior-posterior dimensions of the left auricle and an increase in the left ventricular ejection fraction in ChCI patients. PMID- 10672719 TI - [The effect of enalapril on heart rate variability in patients with arterial hypertension]. AB - Spectral indices were studied for variability of the heart's rhythm (HRV) in patients with arterial hypertension (AH), depending on age, degree of severity of hypertension under exposure to the enalapril maleate therapy. In AH patients, HRV indices suggested high risk of sudden cardial death, especially in those persons presenting with severe AH. Under exposure to enalapril maleate patients with isolated systolic AH, severe and moderately severe AH demonstrated changes in their HRV indices reflecting rise in the activity of the parasympathetic section of the vegetative nervous system, which fact can serve as a positive prognostic index of mortality in AH patients. PMID- 10672720 TI - [Forlax in the treatment of the constipation syndrome in children with combined digestive organ pathology]. AB - Submitted in the paper are data on efficacy of the drug Forlax (Beaufour Ipsen, France) in the treatment of the constipation syndrome in those children presenting with associated pathology of the digestive system. The drug was found out to be effective in children, its therapeutic effect being recordable over one month in 76% of patients, which fact warrants further study of the curative potencies of the drug in children. PMID- 10672721 TI - [The combined treatment of calculous pyelonephritis with the use of perfusion of the kidney cavitary system with the preparation Palisan in the postoperative period]. AB - Results are submitted of use of perfusion of the kidney cavitary system with the drug Palisan in the postoperative period in a multimodality treatment of calculous pyelonephritis. On the basis of bacteriological investigations a high efficacy of the drug in carrying out perfusion of the caliceal-and-pelvic system in treating the inflammatory process in the kidneys has been ascertained. An optimal methodical approach to the carrying out of perfusion has been developed. Also emphasized in the paper is value of perfusion of the kidney cavitary system in metaphylaxis of urolithiasis. PMID- 10672722 TI - [The efficacy of the preparation Fastum-gel in the combined treatment of patients with different clinical syndromes of lumbar osteochondrosis]. AB - As a result of clinical and functional investigations conducted in 47 patients with reflex and compression and radicular syndromes of lumbar osteochondrosis efficacy was substantiated of inclusion into the combined physiobalneological treatment of applications with the drug Factum-gel (Berlin-Chemie, Menarini Group). The author suggests that quantitative electromyographic determinants be used permitting the differentiation between reflex and compression and radicular syndromes and allowing the follow-up to be carried out of those patients presenting with the above clinical syndromes of lumbar osteochondrosis. PMID- 10672723 TI - [The efficacy of amizon in the combined treatment of patients with epidemic parotitis]. AB - Efficacy was studied of a new anti-inflammatory interferon-inducing drug preparation amizon in a combined therapy of epidemic parotiditis in 118 patients versus the group of comparison consisting of 147 patients. A positive effect was ascertained of the drug on clinical parameters, decrease in the incidence rate and in severity of complications (orchitis, pancreatitis), immune indices. PMID- 10672724 TI - [Current aspects of the classification of cholelithiasis and the principles of the therapeutic procedure in its different forms]. AB - A classification is submitted of cholelithiasis, taking account of stages of the illness, its main clinical forms and complications, with the diagnostic criteria and policy of treating it being pointed out. The use of the above classification ensures succession of surgeons' and therapists' work in questions of diagnosis, prophylaxis, and treatment of patients with cholelithiasis. PMID- 10672725 TI - [The classification of cystic lesions of the pancreas]. AB - The paper focuses on different classifications of cystic affections of the pancreas. The most simplified classification is proposed to be used in routine practice, which facilitates clinical recognition and permits the selection of optimum, in the first place, surgical, tactics. PMID- 10672726 TI - [The use of preparations of plant origin in treating and rehabilitating elderly patients with chronic hepatitis]. AB - The article contains data on use of vegetal drugs Hepatofalk Planta, chicory, Achillea millefolium in the treatment and rehabilitation of chronic hepatitis in elderly subjects. PMID- 10672727 TI - [The principles, levels and sources of the financing for the medical care system under the new socioeconomic conditions (a review of the literature)]. AB - A review is submitted of home and foreign literature on the state of financing of the medical industry. The data presented reveal modern principles, levels, and sources of financing of public health care systems in our country and abroad. The experience studied suggest that unification of scientific and practical achievements of Ukraine and economically developed countries, together with further improvements in financing of the medical industry might contribute to formation of national efficiently functioning public health care system. PMID- 10672728 TI - [Morbidity and temporary loss of work capacity as a consequence of arterial hypertension in rural inhabitants]. PMID- 10672729 TI - [The pharmacoeconomics of testicular cancer in Ukraine]. AB - An ever-growing cost of health care in Ukraine attest to the need for an optimum employment of resources in diagnosing and treating disorders of major public health significance, testicular carcinoma (TC) among their number. The authors consider it expedient to ascertain TC costs and design an analytical framework for the diagnostic and therapeutic options to be assessed properly. The following combination chemotherapy regimens were found to be associated with high response rates and acceptable longterm survival: BEP, PE programmes for primary treatment and PEI programme for recurrent or refractory TC. The minimum cost of the above regimens in one course of treatment (with 2 to 4 usually prescribed) for one patient, his height 175 cm, weight 75 kg--S = 1.9 m2, is US$ 613.0, 385.9, 813.5 for BE100P, PE100, and PEI respectively. In the Lviv region with the population about 2,750,000 TC cytostatic therapy is estimated to be about US$ 75,168.6 this being 62.6% of the annual budget appropriation for drug therapy of all malignant diseases taken together. The importance is emphasized of pharmaco-economic principles in the clinical management of patients allowing for decisions to be made with taking account of the cost and expected outcome of the therapy instituted. PMID- 10672730 TI - [The age-related structure of the morbidity of background and precancerous cervix uteri pathology]. AB - For all breakthroughs in the treatment of cancer of the uterine cervix diagnosis of the incipient stages of malignancy still poses unique problems. The studies made on the basis of the clinical hospital N 16 and the Oncologic Centre of the Ministry of Health of Ukraine, histological investigations conducted on the basis of the clinical hospital N 25 and the Ukraine MH Oncological Centre permitted the examination of 209 women who had applied for medical advice for background and pretumour disorders of the neck of the womb. On having done the statistical processing of data, analysis of the secured findings was performed. The choice of the relevant method and theoretical preparation for the regression analysis were put through as recommended by Lakin G. F. Thus, the uterine cervix background and pretumour morbidity has two age peaks of different degree: the first--the main peak--falls on age 25, the second peak--that of a weaker degree--falls on age 38 years. The above two peaks fall on the active reproductive period of life (25 to 40 years of age)--just that time in women's life during which high case rates are recordable with background and pretumour pathology. PMID- 10672731 TI - On the theory of behavioral mechanics. AB - The Theory of Behavioral Mechanics is the behavioral analogue of Newton's laws of motion, with the rate of responding in operant conditioning corresponding to physical velocity. In an earlier work, the basic relation between rate of responding and sessions under two FI schedules and over a range of commonly used session values had been shown to be a power function. Using that basic relation, functions for behavioral acceleration, mass, and momentum are derived here. Data from other laboratories also support the applicability of a power function to VI schedules. A particular numerical value is introduced here to be the standard reference value for the behavioral force under the VI-60-s schedule. This reference allows numerical values to be calculated for the behavioral mass and momentum of individual animals. A comparison of the numerical values of the momenta of two animals can be used to evaluate their relative resistances to change, e.g., to extinction, which is itself viewed as a continuously changing behavioral force being imposed on the animal. This overall numerical approach allows behavioral force-values to be assigned to various experimental conditions such as the evaluation of the behavioral force of a medication dosage. PMID- 10672732 TI - Psychological variables associated with adolescents' identification with significant groups. AB - This research describes associations of identification of 300 adolescents with significant group concepts and variables correlated with their development such as self-esteem, cognitive moral reasoning, and self-perceptions of personality. PMID- 10672733 TI - Holding emotional and linguistic rulers up to the poetry of Robert Frost. AB - The words in a large and representative sample of Robert Frost's poetry were compared with information in several data bases which provided estimates of the poetry's pleasantness, arousal, emotionality, imagery, and linguistic complexity. Findings confirmed that Frost's poetry was linguistically simple and emotionally restrained in comparison to that of his cohort. They also highlighted the increasing variability in Frost's poetry across time and its decreasing imagery. Frost's poetry was more restrained than that of his male cohort but similar to the poetry of his female cohort in its linguistic simplicity. Frost's acknowledged death poems were in fact quite pleasant and passive in emotional tone. PMID- 10672734 TI - Girls' fearfulness as a product of mothers' fearfulness and fathers' authoritarianism. AB - Data from 27 girls of 10 to 12 years were obtained on the Fear Survey Schedule for Children--Revised and from their parents on the Fear Survey Schedule for Adults-III and the California F Scale. Analyses indicated fathers' authoritarianism scores and mothers' fear scores have independent associations with children's fears and suggested greater behavioural overcontrol by fathers and the greater propensity of mothers to communicate threatening information. PMID- 10672735 TI - Psychosocial factors associated with peptic ulcer in aged persons. AB - The relationship between psychosocial factors and the occurrence and aggravation of peptic ulcer was studied in elderly people. Thirty-nine (14 male) ulcer patients and 79 (30 male) elderly people aged 65 years and over and living in the community were tested. Information on health status, medication, lifestyle, and psychosocial status was obtained by means of interviews using a questionnaire and from the clinical records of the ulcer patients. The same questionnaire was distributed to the residents. Logistic regression analysis adjusting for age disclosed that peptic ulcer was significantly associated with having an occupation and low exercise practice in men. However, the relationship weakened below statistical significance after adjusting for some physical risk factors besides age. Low education was significant but low instrumental support fell short of statistical significance with peptic ulcer in women even after adjustment for several physical risk factors besides age. Thus, the specific psychosocial factors might be independently associated with the occurrence and aggravation of peptic ulcer in at least elderly women. PMID- 10672736 TI - Some social correlates of homicide rates in Italy. AB - In Italy, as elsewhere, homicide rates are higher in poorer regions. Unemployment rates associated strongly and positively with homicide rates, explaining up to 54% of variance in regional distribution of homicide rates across the 20 Italian administrative regions. PMID- 10672737 TI - Perceived control, autonomy, and self-regulated learning strategies among Japanese high school students. AB - This study investigated relations among measures of perceived control, autonomy, and self-regulated learning strategies for 228 junior high school (90 in Grade 7 and 138 in Grade 8) and 306 senior high school (184 in Grade 11 and 122 in Grade 12) students. Participants completed three self-report questionnaires designed to measure control beliefs, strategy beliefs, capacity beliefs, seven types of motivation, and two types of self-regulated strategies. Confirmatory factor analysis identified the structure of perceived control modeled by Skinner, Chapman, and Baltes (1988), the seven-factor structure of autonomy by Vallerand, Pelletier, Blais, Briere, Senecal, and Vallieres (1992, 1993), and the two types of self-regulated learning strategies by Pintrich and De Groot (1990). Significant "grade" differences were obtained in several measures. Canonical correlation was used to investigate the relations between perceived control and autonomy measures. Finally, multiple regression analysis was used to investigate the relations between perceived control and self-regulated learning strategies and between autonomy and self-regulated learning strategies. Implications of the results are presented. PMID- 10672738 TI - Obsession-compulsion in college students in the United States and Kuwait. AB - American (n = 132) and Kuwaiti (n = 204) undergraduates responded to the Arabic Obsessive-Compulsive Scale in English and Arabic languages, respectively. Students in Kuwait scored as more obsessive-compulsive than their American counterparts. PMID- 10672739 TI - Job satisfaction and intent to continue working among individuals with serious mental illness. AB - This study investigated the relationship between job satisfaction and the intention to continue working in a sample of 87 individuals with psychiatric disabilities who worked in supported or supervised employment. Data were collected from respondents in Columbus, Ohio in 1996. After controlling for selected correlates (education, length of employment, contract type, social support, and attitudes concerning pay and toward work), standard multiple regression analyses indicated that job satisfaction was a significant predictor of intent to continue working. Analyses also indicated that respondents who completed high school and who said that they worked primarily for the money were less likely to want to continue working at current jobs. These findings may be used to improve the vocational success of individuals with psychiatric disabilities who work in supported and supervised employment. PMID- 10672740 TI - Methodological note on use of the Center for Epidemiological Studies--Depression Scale with older samples. AB - Samples of older adults (ns = 600, 251, and 214) given the Center for Epidemiological Studies--Depression Scale by telephone seem to have composite scores that are much lower than those who are interviewed face-to-face. PMID- 10672741 TI - Religiosity and pathology. AB - In a sample of 109 undergraduate students, religiosity was not associated with the fear of death or with scores on a manic-depressive tendencies scale. PMID- 10672742 TI - Forming clinical impressions during the first five minutes of the counseling interview. AB - The influence of earlier formed clinical impressions of the client on later judgments within a counseling session were examined. It was predicted that redundancy in judgments of counselors' clinical impressions of the client after 5 min. and after 30 min. of an interview would be greater for counselor-trainees who are (a) more self-confident about their judgments and (b) lower in cognitive complexity, than other counselor-trainees. A stimulus tape in which a male counselor conducted an initial 30-min. interview with a depressed female client was shown to 80 counselor-trainees. At the 5- and 30-min. marks, the tape was stopped, and the counselor-trainees indicated their clinical impressions of the client. These impressions included written thoughts and ratings of clinical characteristics of the client. Analyses showed that both their self-confidence and cognitive complexity were not associated with differences in redundant judgments. PMID- 10672743 TI - Effect of reduced immunity on murine behaviors. AB - Reduction of immunity in 11 mice of the CD1 strain by administration of methotrexate did not alter some reliable behaviors in the open-field (ambulation, rearing, defecation) compared with 10 matched mice injected with saline. Perhaps moderate depression of immunity may not affect some characteristic behaviors. PMID- 10672744 TI - Beliefs in the paranormal: age and sex differences among elderly persons and undergraduate students. AB - Beliefs in the paranormal were rated stronger in younger as compared to elderly adults by Emmons and Sobal in 1981, and sex correlates of paranormal beliefs appeared to be stronger in women than in men by Irwin in 1994. This research studied possible linkages between age and sex with a comparative analysis between results of Vitulli and Luper's 1998 survey among undergraduate students and data from elderly men (M = 72 yr., SD = 9.2, n = 21) and women (M = 69.3 yr., SD = 7.7, n = 55). Crawford and Christensen's 1995 12-item Extrasensory Perception Survey was administered to elderly persons living in apartment complexes and private homes, participating in activities in a recreation center, or attending a continuing-education seminar. A 2 x 2 multivariate analysis of variance from responses on the 12-item survey showed that undergraduate men and elderly women had the highest ratings on paranormal beliefs. The self-selecting characteristics of a segment of the elderly sample led to a post hoc univariate analysis of variance by partitioning that sample into those who were attending a continuing education seminar versus all other elderly persons. Summated ratings (total scores) for this survey showed main effects for these subsamples and for sex. Sex and age differences were discussed in the context of the hypothesis of social marginality. PMID- 10672745 TI - Domestic social integration and suicide in France. AB - In 95 French Departements in 1970, suicides rates were higher where measures of domestic social integration (marriage and divorce rates) were greater. PMID- 10672746 TI - Chronological age and performance of persons with mental retardation on verbal subtests of the Wechsler Intelligence Scale for Children--Revised, French Version. AB - The Influence of chronological age on verbal intelligence of persons with metal retardation was studied using the French version of the Wechsler Intelligence Scale for Children--Revised. Participants were 1,101 children and adolescents divided into two groups according to the severity of metal retardation. The first group included 551 participants with moderate mental retardation, the second 550 participants with mild mental retardation. Analyses indicated a significant association between chronological age and raw scores on the four Verbal Comprehension subtests, even with performance on Perceptual Organization and Freedom from Distractibility subtests held constant. This finding suggests the relationship between chronological age and verbal intelligence is valid for persons with moderate mental retardation as well as for those with mild metal retardation. PMID- 10672748 TI - Panel-touch behavior of horses established by an autoshaping procedure. AB - Panel-touch behavior of 3 geldings was successfully established by a response termination type of autoshaping procedure. An omission or negative contingency introduced after the training of an animal, however, decreased the response rate to a near-zero level. PMID- 10672747 TI - Correlations between parents' and teachers' ratings of social skills for a group of developmentally disabled children in Iran. AB - Parents' and teachers' ratings of social skills and behavioural problems for 89 special education students aged 8 to 15 yr. were measured using the Social Skills Rating System of Gresham and Elliott. The sample was selected in a school for educable mentally retarded children in Shiraz, Iran. The low to moderate correlations between the two sets of ratings suggest that assessment of social skills and behavioural problems should include the use of different rating scales in more than one setting. Sex differences were not significant for parents' and teachers' ratings of these disabled children. The scores showed high internal consistency. PMID- 10672749 TI - Economic consequences and lack of respite care. AB - The study examined the influence of a lack of availability of respite care on the careers of 574 parents having children with disabilities. Among these parents, there was a relationship between age of child, severity of disability, and parents missing hours of work or passing up occupational opportunities. These findings have implications for advocating more available and accessible respite services and more in-depth study of the cost effectiveness of respite care on parental income and career progression. PMID- 10672750 TI - Children and electronic games: a comparison of parents' and children's perceptions of children's habits and preferences in a United States sample. AB - Despite the popularity of violent electronic games, anecdotal evidence suggests that many parents lack even basic information about children's game-playing habits. The goal of the present study was to examine parental knowledge of children's electronic game-playing habits by assessing the congruence between children's and parents' perceptions of child's playing time, parental supervision, game preference, and reaction to actual game-playing. 35 children in Grades 3 to 5 and one parent of each child completed a background questionnaire and played either a violent or nonviolent electronic game. In paired comparisons, parents reported significantly higher estimates of supervision time than children. Most parents either named an incorrect game or were not able to guess their child's favorite game. In 70% of these incorrect matches, children described their favorite game as violent. Parents may underestimate their child's exposure to violence in electronic games. After playing the same electronic game as part of the study, parents reported significantly higher frustration than children. Higher frustration with game-playing could contribute to deficits in parental knowledge of children's playing habits. PMID- 10672751 TI - The Depression-Happiness Scale: test-retest data over two weeks. AB - The present aim was to examine further the psychometric properties of the Depression-Happiness Scale. Test-retest data over 2 wk. are reported for this scale for a sample of 54 female Northern Irish university students. Stability was fairly high as r = .70, and there was no significant change between Time 1 and Time 2 (M = 49.7 and 50.8, respectively). The data suggest that the Depression Happiness Scale might be considered a trait measure of happiness rather than a state measure. PMID- 10672752 TI - Attitudes toward abortion, capital punishment, and assisted suicide. AB - In 51 college students, attitudes toward abortion, capital punishment, and assisted suicide were not consistently anti- or pro-death. PMID- 10672753 TI - Smokeless tobacco use among urban white and black South Africans. AB - A telephone survey was conducted to compare the extent of smokeless tobacco use and perception of related health risks by white and black urban South Africans. Using systematic random sampling, one out of every 20 phone numbers was selected from the Seshego (blacks) and Pietersburg (whites) telephone directory until 300 tobacco users in each site were identified. Among the white group, cigarette smoking was clearly predominant (290) and only 10 used snuff, whereas among the black sample almost half (46.7%) of the tobacco users used snuff, especially women (40%). Although a majority acknowledged negative effects of snuff use on their health and its addictive character, 42% either do not believe or do not know that snuff contains nicotine and causes cancer. PMID- 10672754 TI - Factor analysis of the continuous performance test and parent-teacher reports of attention deficit disorder. AB - The relationships between a computerized measure of attention deficit disorder and scores from two commonly used parent-teacher reports were investigated. A factor analysis of the raw omission and commission scores provided by the Continuous Performance Test and Conners' Parent Rating Scale and the ADD-H Comprehensive Teacher Rating Scale indicated that for a sample of 54 children the Continuous Performance Test was most closely associated with measures of impulsivity and hyperactivity provided by the Conners' rating. This finding was congruent with the use of the Continuous Performance Test in the evaluation as a measure of Attention Deficit Hyperactivity Disorder and suggestive of a positive and significant relation between this computerized measure of behavior and parents' perception of behavior. Little association was detected between scores on the teachers' scale and omission and commission scores. PMID- 10672755 TI - Commentary on the possibility that Viagra may contribute to transmission of HIV and other sexual diseases among older adults. AB - Many older male adults experiencing impotence are being treated with Viagra. Scientific and media reports indicate that this medication is effective in re establishing sexual relationships among these men. Despite the benefits that Viagra may have in the treatment of impotence among older adults, increased sexual activities could also facilitate the spread of HIV infection and other sexually transmitted diseases. This holds for older adults who may lack factual knowledge of HIV transmission and perceive themselves as nonsusceptible to this disease. Prescription of Viagra in combination with HIV/AIDS prevention programs may be helpful. PMID- 10672756 TI - A new beginning in application of Frankl's work to emerging issues in psychotherapy. AB - Recent interest in Frankls' system of thought, supported by Langle, paves the way for its critical and fruitful appropriation in psychotherapy and existential analysis. PMID- 10672757 TI - Relation of functioning with beliefs about coping, and future time perspective. AB - 51 college students were administered the Reasons for Living Inventory, Zimbardo's Time Perspective Inventory, and the Environmental Deprivation Scale Questionnaire. Pearson product-moment correlations indicated that those functioning well (low scores on the Environmental Deprivation Scale Questionnaire) tended to have high scores on the future subscale of the Zimbardo Time Perspective Inventory (r = -.45, p < .001). There were no significant correlations with the Reasons for Living Inventory's total score or its Survival and Coping Beliefs subscale (rs = .01 and .05, respectively). The results suggest the use of multidimensional cognitive and behavioral data to guide intervention to improve one's level of functioning. A limitation of the study is the small number of participants. PMID- 10672758 TI - The Leiden Quality of Work Questionnaire: its construction, factor structure, and psychometric qualities. AB - Based on two leading models in occupational stress research, the Job Demand Control-Support model and the Michigan model, a comprehensive quality of work questionnaire, was constructed--the Leiden Quality of Work Questionnaire. The factor structure of this questionnaire was assessed and cross-validated in two sub-samples of 2,000 men and women from a large sample of the Dutch working population. Analysis indicated that the questionnaire measures 11 work characteristics of Skill Discretion, Decision Authority, Task Control, Work and Time Pressure, Role Ambiguity, Physical Exertion, Hazardous Exposure, Job Insecurity, Lack of Meaningfulness, Social Support from Supervisor and Social Support from Coworkers, and the outcome variable of Job Satisfaction in a reliable way. PMID- 10672759 TI - Alexithymic characteristics of HIV-positive patients. AB - Despite extensive psychiatric research on HIV-positive patients, there are no published studies on alexithymia. Alexithymic characteristics and related factors were examined in a sample of 81 HIV-positive patients using the modified Beth Israel Psychosomatic Questionnaire and the Social Support of Stress and Coping Inventory. The severity of alexithymia was significantly higher in HIV patients than healthy controls, suggesting the presence of secondary alexithymia. Scores on two alexithymic characteristics, affect awareness and operational thinking, significantly correlated with ratings of poor utilization and perception of social support. As the severity of HIV infection progressed, affect awareness was higher, controlling for poor utilization and perception of social support. These results suggest that secondary alexithymia, associated with poor utilization and perception of social support, may be manifest as a state reaction to approaching death. PMID- 10672760 TI - Relationship between belief in good luck and general health. AB - 62 undergraduate university students were administered the 12-item Belief in Good Luck Scale of Darke and Freedman and the General Health Questionnaire of Goldberg and Williams. Scores on belief in good luck showed a significant correlation of .29 with anxiety and -.35 with depression but correlations were not significant for somatic symptoms (.15) and social dysfunction (.15). PMID- 10672761 TI - A cross-cultural evaluation of depression in children in Egypt, Kuwait, and the United States. AB - The English version of the Arabic Children's Depression Inventory, constructed by Abdel-Khalek, was applied to a sample of 535 U.S. students (11 to 18 years old). Cronbach coefficients alpha were .88, .90, and .89 for boys, girls, and all subjects, respectively. Seven factors were extracted by principal axis factor analysis (Negative mood and self-depreciation, Fatigue, Lack of loneliness, Sleep problems, Weak concentration, Pessimism, and Feeling happy), denoting clear factorial structure; however, the scale was intended to be unidimensional. Sex and racial differences for this American sample were not statistically significant but the correlation of depression scores with age was .22. The scale appears useful in studying depression in American school children and adolescents. Also, cross-cultural differences in childhood depression between samples from Egypt and Kuwait of previous studies and the present American sample were examined. Based on the effect size, female Kuwaiti had a lower mean depression score than either the Egyptian or American groups. The scale can be used in cross-cultural research. PMID- 10672762 TI - Job stress among general practitioners and nurses in primary care in England. AB - This is a description comparing job stress, job satisfaction, and mental well being of general practitioners (n = 205) and practice nurses (n = 119) in England, based on responses to a questionnaire. General practitioners reported lower job satisfaction and significantly greater pressure at work than did the practice nurses. Also, male general practitioners had significantly higher scores on anxiety and depression than a British normative population. Practice nurses, on the other hand, reported lower scores on anxiety and depression. The results should be interpreted with caution as the study is based on a small sample limited to the northwest region only; however, it does provide information which has important implications for the well-being of doctors and nurses in primary care. PMID- 10672763 TI - Three screening tests for patients with psychological problems who undergo plastic and reconstructive surgery. AB - To investigate their quality of life, screening tests of emotion, coping with stress, and personality were developed for use with patients in plastic and reconstructive surgery who had psychological problems. The validity and reliability of each test were examined in a sample of 329 patients who were admitted to the Department of Plastic and Reconstructive Surgery. The reliability of each test was supported by factor analysis, adequate internal consistency, and test-retest correlations. Also, validity was acceptable. Although further examination is required, these screening tests seem suitable for use in research among this type of patient in Japan. PMID- 10672764 TI - Initial evaluation of the Secondary Trauma Questionnaire. AB - Many measures exist to evaluate posttraumatic stress disorder (PTSD), but there are few ways of assessing secondary traumatic stress disorder and these are limited to specific populations. Secondary traumatic stress disorder involves the transfer of trauma symptoms from those who have been traumatized to those who have close and extended contact with trauma victims. Thus, family members of those who have been traumatized and therapists who treat trauma survivors are vulnerable to developing secondary traumatic stress disorder. In this initial evaluation of the newly developed Secondary Trauma Questionnaire, 261 mental health professionals and 157 college students were evaluated. Analysis indicated that the questionnaire showed good internal consistency and was significantly correlated with known measures of trauma. The Secondary Trauma Questionnaire is presented as a promising way to measure secondary trauma symptoms and further research using this questionnaire appears to be warranted. PMID- 10672765 TI - Changes in women's fear of success and fear of appearing incompetent in business. AB - The primary purpose of this research was to examine whether fear of success and of appearing incompetent among women have changed recently. Another purpose was to examine whether such fears differed among women who hold Traditional views and those who hold Progressive views about the roles of women in the workplace. The Fear of Success Scale, the Fear of Appearing Incompetent Scale, and the Attitude Toward Women Scale were completed by 61 male and 52 female graduating seniors. Significant differences were found between the groups for scores on the Attitude Toward Women Scale, but none between the sexes for scores on the Fear of Success Scale or the Fear of Appearing Incompetent Scale. Significant differences were found, however, on the latter two scales when women were separated into Traditional and Progressive groups. PMID- 10672766 TI - Relationship between properties of words and elicitation of skin conductance response. AB - We investigated the association of subject-rated imagery, subject-rated concreteness, subject-rated emotionality, frequency, date of entry into the language, and word length with emotional imagery as measured by the skin conductance response elicited by that word. 50 words in a list of 25 word-pairs were rated by 96 university students; then their skin conductance response of each word was measured for each word. In each pair, one word was concrete and one was abstract but with related meaning, e.g., adolescent and adolescence. Stepwise multiple regression indicated that 30% of variance in the later skin conductance response was explained by imagery and subject-rated emotionality. Imagery alone explained 24% of variance. PMID- 10672767 TI - Accurate knowledge about suicide. AB - Accurate knowledge about suicide among 150 English students was not associated with age, sex, personality, or attitudes toward suicide. PMID- 10672768 TI - Locus of control as a dispositional determinant of men's traditional sex-role attitudes. AB - The associations among measures of locus of control and attitudes toward women who work were assessed using data from the National Longitudinal Survey of Youth for 1,229 young adult males. Significant positive correlations, ranging from .09 to .14, were found among locus of control and men's negative stereotypes of women who work, which suggests that males who feel a lack of personal control may oppose women working outside of the home. The results also indicate that men with greater personal control may be more accepting of women in nontraditional sex roles than men with an external locus of control. Negative stereotypes of women who work and male self-preservation may explain these relationships. PMID- 10672769 TI - Description of older adults as depicted in magazine advertisements. AB - Negative attitudes about aging have been widespread and films, television, radio, and print media may serve as an important source of socialization or reflect the current views of older adults. This study focused on examination of the frequency of depictions of older men and women in 765 advertisements appearing in Time and Newsweek national weekly news magazines, and on an analysis of their roles suggested in photographs depicting a total of 2,505 persons. These were collected over a one-year period and coded by three persons. Analysis indicated that older adults, especially older women, were not only presented infrequently but, when presented roles, were often passive or dependent as is consistent with social stereotypes. PMID- 10672770 TI - Comment on "manic-depressiveness and its correlates". AB - Data from a sample of 612 college students showed that the Depression subscale of the Manic-Depressiveness Scale had greater internal reliability than the Mania subscale. Furthermore, correlates of the total scale score did not appear to provide as useful information as use of the Depression and Mania subscales separately. PMID- 10672771 TI - [Apocalypse 2000: the end of science and the golden age of pseudo-medicine]. PMID- 10672772 TI - [Digestive emergencies of elderly patients]. AB - Evaluation of an elderly patient with acute abdominal conditions presents a challenge to the physician on account of the general poverty of history and clinical signs and the poor reliability of diagnostic procedures. The management of these pathologies are analyzed: cholecystitis, appendicitis, incarcerated hernia, small bowel obstruction, colonic diverticular disease, intestinal ischemia and gastroduodenal ulcer. An early elective treatment of chronic pathologies like gallstones, hernias and intestinal ischemia trends to avoid complications. PMID- 10672773 TI - [Autosomal dominant spinocerebellar ataxia]. AB - The autosomal dominant spinocerebellar ataxias (SCA) are a heterogeneous group of degenerative diseases presenting with ataxic gait, limbs ataxia, dysarthria and cerebellar oculomotor disturbances. Usually, cerebellar signs are associated with pyramidal signs, extra-pyramidal signs, spinal signs and signs of peripheral neuropathy. Neuropathological studies have disclosed an involvement of the cerebellum and its afferent/efferent pathways, of the brainstem and of the spinal cord. Distinct entities are now recognized: SCA1, SCA2, SCA3/Machado-Joseph disease, SCA4, SCA5,SCA6, SCA7 and dentatorubropillidoluysian atrophy (DRPLA). In most cases, a CAG trinucleotide repeat expansion has been demonstrated by genetic investigations. Moreover, recent studies have shown that autosomal dominant spinocerebellar ataxias are characterized by intra-nuclear inclusions containing polyglutamine in affected cells. These complexes might pl ay a determinant role in the neurodegenerative process. Cell death could be due to accumulation of a polyglutamine as a result of trinucleotide repeats. PMID- 10672774 TI - [Adenopathy and eosinophilia]. AB - The exercise topic is the persistence of adenopathies for many years in a young marocan boy who also presents with peripheral blood eosinophilia. PMID- 10672775 TI - [Reboxetine (Edronax)]. AB - Reboxetine is a new antidepressant acting by selective inhibition of noradrenaline reuptake (NARI) at the synaptic cleft. The efficacy of reboxetine is similar to other antidepressants such as tricyclic and Selective Serotonin reuptake inhibitors once (SSRI). A faster onset of action (in a mean of 10 days) is suggested by preliminary studies. Reboxetine seems also to improve social adjustment of the treated depressed patients. Due to its selectivity, reboxetine presents a favourable safety profile. Treatment of depression will certainly benefit from this new compound. However, some issues need to be clarified such as the use of reboxetine in combination with other antidepressants. PMID- 10672776 TI - [The search for medical information on the World Wide Web]. AB - The internet has experienced tremendous growth over the past few years and has currently many resources in the field of medicine. However, many physicians remain unaware of how to gain access to this powerful tool. This article briefly describes the World Wide Web and its potential applications for physicians. The basics of web search engines and medical directories, as well as the use of advanced search with boolean operators are explained. PMID- 10672777 TI - [Ventricular repolarization anomalies in an asymptomatic patient]. PMID- 10672778 TI - [Albert Brachet, the "Standard of Anatomy"]. PMID- 10672779 TI - Exposure to environmental tobacco smoke and the risk of lung cancer: a meta analysis. AB - A meta-analysis was carried out to calculate a pooled estimate of relative risk of lung cancer following exposure to environmental tobacco smoke (ETS) and to determine whether there was any heterogeneity in the pooled estimates according to selected characteristics of the studies. A total of 35 case-control and five cohort studies providing quantitative estimates of the association between lung cancer and exposure to ETS published between January 1981 and March 1999 were identified. Using fixed- and random-effects models, we calculated pooled estimates of relative risk for exposure to ETS from subjects' parents (during childhood), spouses, and coworkers. As well, we investigated whether the pooled estimates of relative risk varied by study location, degree of control of potential confounding variables, proportion of cases confirmed histologically, proportion of surrogate respondents, nonresponse rates, and year of publication. The relative risk of lung cancer among non smoking women ever exposed to ETS from their husbands' smoking was 1.20 (95% confidence interval (CI): 1.12-1.29). The pooled relative risk was 1.19 (95% CI: 1.10-1.29) for case-control studies and 1.29 (95% CI: 1.04-1.62) for cohort studies. In various subgroup and meta regression analyses, we found no statistically significant differences by selected characteristics of the studies. In addition, we found that the risk of lung cancer increased consistently with increasing levels of exposure. The 11 studies reporting relative risks among male non smokers yielded a pooled relative risk of 1.48 (95% CI: 1.13-1.92) for ever exposed to ETS, and the relative risk of lung cancer for ever being exposed to ETS at work was a 1.16 (95% CI: 1.05 1.28). These results are consistent with the hypothesis that exposure to ETS increases the risk of lung cancer. While there may be alternative explanations to the data, it is more likely that the observed association is not an artifact and that ETS causes lung cancer in non smokers. PMID- 10672780 TI - Prognostic impact of cathepsin B and matrix metalloproteinase-9 in pulmonary adenocarcinomas by immunohistochemical study. AB - The expression of Cathepsin B (CB) and matrix metalloproteinase-9 (MMP-9) in extirpated tissues of adenocarcinomas in non-small cell lung cancer from 90 cases was investigated immunohistologically, and the correlations between the extent of the expression and the clinicopathological features were assessed for investigating the process of tumor metastasis. It is important to reveal the mechanisms of destruction of the basal membrane and infiltration of tumor cells at the primary lesion. Sections were obtained from 10%-formalin-fixed and paraffin-embedded tissues. They were reacted with an anti-human CB polyclonal antibody or an anti-human MMP-9 polyclonal antibody. Of 90 patients, 58 (64.4%) and 39 (48.3) cases were found to be positive for CB and MMP-9 expression, respectively. A significantly higher extent of the CB expression was observed in the tissues of patients who showed postoperative recurrence of the tumor (P = 0.013). Especially, a similar observation was obtained among early cases of T1N0 (P = 0.023). In contrast, no such tendency was demonstrated in the expression profile of MMP-9. Furthermore, the enzyme expressions were compared among different types of metastases. Patients with higher extents of CB expression tended to show significantly higher rates of hematogenous and intrapulmonary metastases (P = 0.023 and P = 0.010, respectively). However, there was no significant correlation between MMP-9 expression and the prognostic factor of the patients. Therefore, we suggested that evaluation of CB expression in the tumor tissue might be useful as a postoperative prognostic factor of pulmonary adenocarcinoma. Especially, early cancer of T1N0 cases showing higher expression of CB may need postoperative adjuvant chemotherapy. PMID- 10672781 TI - Immune cell infiltrates and prognosis in primary carcinoma of the lung. AB - The prognostic significance of immune cell infiltrates in surgically resected human lung cancer was investigated in 710 patients. Lymphoid infiltrates were quantified on both standard H&E stained sections and, in a subset of 95 cases, using immunohistochemistry and antibodies to CD3, CD8, CD57, CD68, CD79a and S100 to identify various immune cell types. Subjective grading (low, moderate, high) of lymphoid cell infiltrates on H&E sections of tumour and measurement, using image analysis, of overall level of tumour infiltration by any of the immunohistochemically labelled specific immune cell types of the stained sections showed no prognostic significance. However, when a distinction between peritumoural and intratumoural infiltration by particular cell types was made, intratumoural infiltration by high levels of CD3+ and S100+ cells was associated with longer post-operative survival (P = 0.02 and P = 0.045, respectively). In lung cancer, subjective assessment of tumour lymphoid infiltration and overall levels of infiltration by particular immune cell types carries no prognostic significance. Intratumoural infiltration by relatively high numbers of CD3+ T lymphocytes and Langerhans cells (S100+) is associated with a better patient outcome. PMID- 10672782 TI - Phase II trial of induction high-dose chemotherapy followed by surgical resection and radiation therapy for patients with marginally resectable non-small cell carcinoma of the lung. AB - The combination of carboplatin and paclitaxel is an active regimen in non-small cell lung cancer (NSCLC). Historically, patients with stage III disease have manifested higher response rates than patients with metastatic disease, and patients achieving a pathologic complete response to induction chemoradiation therapy prior to surgery have shown better long-term outcome. Based upon our pilot data using high-dose carboplatin and paclitaxel, we designed a phase II trial in patients with marginally resectable stage IIIA NSCLC. Ten patients, with bulky nodal stage IIIA disease, initially received etoposide (2 g/m2) and granulocyte colony-stimulating factor (G-CSF) to mobilize peripheral blood stem cells (PBSC). Two cycles, 28 days apart, of carboplatin (AUC 12 in seven patients; AUC 16 in three patients) and paclitaxel (250 mg/m2) were administered with filgrastim (5 microg/kg) and PBSC support. After re-evaluation, patients underwent a thoracotomy followed by radiotherapy (44-60 Gy) if deemed resectable, or radiotherapy alone (60 Gy) if not resectable. The median age was 58.5 years (48-66) with a median ECOG performance status of 0 (0-1). Histology was adenocarcinoma in seven patients; the remainder had either squamous cell, large cell or bronchoalveolar carcinoma. Based on CT radiography, the overall response rate was 40%. Eight of ten patients underwent resection with four right pneumonectomies, three right upper lobectomies and one wedge resection of the right upper lobe. Six patients had a complete resection. Of eight patients resected, four were downstaged by induction therapy, three remained unchanged and one was found to have more extensive disease. The remaining two patients developed metastatic disease while receiving chemotherapy. The median dose of postoperative radiotherapy was 54 Gy (35-66 Gy). Actual median follow-up for all patients was 89 weeks (25 to 136+). The actuarial median overall survival was 124 weeks (25 to 136+) and time to progression was 57 weeks (17 to 136+). The median dose of carboplatin delivered expressed as mg/m2 was 779 (615-1540). Neutropenic fever occurred in two patients during the initial mobilization cycle only. The median number of units of RBC and/or platelets transfused was 0 (0-2 and 0-6, respectively). There were no significant non-hematologic toxicities. High-dose induction chemotherapy with stem cell rescue is feasible and safe with an acceptable response rate. Thoracotomy, including pneumonectomy and postoperative radiotherapy, were well tolerated by patients after undergoing high-dose induction chemotherapy with no apparent increase in peri-operative morbidity. The pathologic complete response rate was low--one out of ten patients. These results indicate that dose escalation of induction chemotherapy does not improve response rates even in this highly selected patient population. Accordingly, the complexity and potential toxicity of high-dose chemotherapy, as delivered in this trial as neoadjuvant treatment of non-small cell lung cancer, is not warranted. PMID- 10672783 TI - Second-line treatment with gemcitabine and vinorelbine in non-small-cell lung cancer (NSCLC) cisplatin failures: a pilot study. AB - PURPOSE: This pilot study was designed to evaluate the efficacy and toxicity of the gemcitabine/vinorelbine combination in non-small-cell lung cancer (NSCLC) patients who had failed cisplatin-based first-line chemotherapy. PATIENTS AND METHODS: Eligible patients had refractory or resistant NSCLC, WHO performance status 0-2, adequate hematologic parameters and normal hepatic, renal and cardiac function. Gemcitabine (1200 mg/m2) was administered on days 1, 8 and 15; vinorelbine (25 mg/m2) was administered on days 1 and 8, every 4 weeks until patients experienced disease progression. RESULTS: From September 1997 to March 1998, 16 patients were enrolled (six: stage IIIB: ten: stage IV). All 16 patients were assessable for toxicity and evaluated for response. One complete (6.25%) response and no partial responses were observed. Median survival was 25 weeks (95% CI 19-30). A median of 3.31 courses per patient was administered, and the median interval between courses was 28 days. The median delivered dose was 664.85 mg/m2 per week of gemcitabine and 10.71 mg/m2 per week of vinorelbine, corresponding to a relative dose intensity of 0.73 and 0.85, respectively. Grade 2/3 thrombocytopenia occurred in 13 (24.52%) of 53 courses administered. Neutropenia Grade 2/3 occurred in 14 courses (26.41%). There were seven (13.20%) episodes of fever related to the drug administration. Mild asthenia was observed in six (37.5%) patients. Other toxicities were mild to moderate. CONCLUSION: These results suggest that this gemcitabine/vinorelbine combination is not an active salvage regimen in patients with refractory NSCLC. The combination merits further evaluation with modified regimens. PMID- 10672784 TI - Primary primitive neuroectodermal tumor of the lung: report of two cases. AB - Two cases of primitive neuroectodermal tumor of the lung are reported. The first case is a 41-year-old man with a tumor in the left upper lung, and the second case is a 30-year-old woman with a tumor in the right lower lung. In both cases, the tumors originated in the lung but not in the chest wall. No distant metastasis was detected. In case 1, transcutaneous fine-needle biopsy (TCNB) revealed small round cell proliferation, although bronchoscopic examination showed no abnormal findings. Both the expression of Mic2 protein and t(11;22)(q24;q12) translocation were proved in the tumor cells. The tumor cells were positive for periodic acid-Schiff (PAS), neuron-specific enolase (NSE), and vimentin, but negative for Leu7, chromogranin A, and pro-gastrin-releasing peptide (ProGRP). In case 2, bronchoscopic examination showed only compressive change in right lower lobe bronchi. TCNB revealed small round tumor cells expressing Mic2 protein. The tumor cells were negative for leukocyte common antigen, S100 protein, pankeratin, chromogranin A, and desmin, but weakly positive for NSE and moderately positive for Ki-67 (MIB1). Both patients were successfully treated by the combination of surgical resection and chemotherapy, and are alive with no sign of recurrence for approximately 22 months in case 1 and 16 months in case 2. PMID- 10672785 TI - A pharmacokinetic model for multiple sites discontinuous gastrointestinal absorption. AB - PURPOSE: To build a pharmacokinetic model taking into account a discontinuous absorption along the gut, from n successive sites, a non-absorbing intestinal segment being always in between two successive sites. To solve the mathematical model linked with the pharmacokinetic model to obtain the concentration and contribution of each site to absorption, area under curve and bioavailability. METHODS: Whatever the number n of sites, we obtained the Laplace transform of amounts of drug in each site, then plasma concentration, so that concentration or AUC were expressed analytically. When only two absorption sites are present, concentration is obtained from Heaviside's theorem; but for n> or =3, Bromwich's theorem is necessary, a pole being of the order of more than two. RESULTS: Simulations performed with data gathered from the literature allow to find, with n=2 sites, the particular case used for ranitinine and to show the efficacy of each site. For n=3 sites, real data exhibiting three peaks of various magnitude were fitted on our model. CONCLUSION: This general discontinuous oral absorption pharmacokinetic model may be taken as a possible tool to characterize each site of absorption and to estimate the area under curves or bioavailability. PMID- 10672786 TI - Two methods for the measurement of voluntary contraction torque in the biceps brachii muscle. AB - Appropriate measurement of maximal voluntary contraction force of a single limb muscle or of a muscle group is important in clinical and research situations. To measure muscle force, one segment of an isometric measuring brace is fixed to a support and force is applied to the other. The output of this arrangement is affected by additional contributions such as pushing or pulling with the whole body. This paper quantifies the differences between measurements of torque produced by the biceps brachii with the brace fixed versus those produced when the elbow joint was isolated by suspending the brace from cables. No statistically significant differences were found between MVC values observed with the two methods within subjects. However, a statistically significant difference in EMG fatigue indices was observed and attributed to a different sharing of force production among different muscles in the two conditions. We conclude that different brace arrangements may lead to the same maximal force but to different rates of myoelectric manifestations of muscle fatigue since the effort may be shared differently among the muscles of synergic or stabilizing groups. PMID- 10672787 TI - Ultrasound velocity and attenuation in relation to maximum trabecula stress in the patella. AB - The ultrasound velocity and attenuation were examined in 16 sets of human patellae. The average ultrasound velocity of patella was shown to be greater in the superior/inferior direction than in the anterior/posterior and medial/lateral directions. The distribution of bone mineral density (BMD) was also examined. The BMD of the patella varied with location. BMD values were largest at the superior and lateral regions and decreased inferiorly and medially. A two-dimensional finite element analysis was performed on each patella. The maximum von Mises stress occurred along the cortical shell on the non-articular surface. The trabecular von Mises stress existed in the posterior region of the patella. Correlation study showed that patellar BMD was significantly associated with each of three directional ultrasound velocities. The relationship between BMD and ultrasound attenuation was not significant. It was also found that the ultrasound velocity and attenuation were not significantly correlated with the maximum von Mises stress. PMID- 10672788 TI - Effect of aneurysmectomy on left ventricular shape and function: case studies. AB - The three dimensional (3D) conformational changes in three patients with large anterior aneurysm in the left ventricle (LV) were examined before and two years after aneurysmectomy by using 3D Cine-computerized tomography (CT). Endocardial and epicardial tracings of 6-9 short axis images encompassing the entire LV were used to reconstruct the LV in 3D. Thickness and percent thickening were calculated using our 3D-volume element approach. A regional wall stress index (stress/pressure) was calculated from regional curvature and thickness. The analysis showed that following resection of the aneurysm the end-diastolic volume was reduced from 257+/-39 to 183+/-39 ml, end-systolic volume from 172+/-39 to 92+/-46 ml and, ejection fraction increased from 34+/-7 to 51+/-13%. The endocardial aneurysm area decreased from 19.7+/-15.9 to 10.1+/-6.5 cm2, whereas the normal zone area was minimally reduced from 87.4+/-17.6 to 79.8+/-10.8 cm2. The percent thickening of the normal zone increased significantly. It is documented here for the first time by detailed 3D analysis that the resection of the LV aneurysm reduces the aneurysmal area and LV size and improves the global and regional function of the remote normal zone. Therefore, the 3D approach can help to design better surgical technique for this complex operation. PMID- 10672789 TI - Analysis of volition latency on antisaccadic eye movements. AB - The antisaccadic paradigm can be applied to test the suppression of reflexive saccades and the activation of volitional saccades simultaneously. The impaired frontal cortex has been shown to have difficulty in suppressing reflexive saccade (prosaccade) to make a successful antisaccade. Degraded antisaccade performance can also be observed in patients with Parkinson's disease (PD). The studies of PD based on the prosaccadic and antisaccadic paradigms have shown controversial findings; the latency between patients and age-matched controls could be either with or without significant difference. Even with this inconsistency, our previous study and recent analysis have supported that the latency of both prosaccade and antisaccade increases significantly for patients with PD. The objective of this study is to investigate whether prolonged antisaccade latency is caused by the affected volitional decision process (volition latency) or simply by the delayed initiation of saccade with direction opposite to the cue, by measuring prosaccade and antisaccade latency from the intermingled paradigms. Eleven mildly affected patients with idiopathic PD and eight age-matched normal subjects were tested in this study. As compared to the age-matched control, the results showed that prosaccade, antisaccadic, and volition latency of the patients was significantly elevated (P<0.01). We conclude that antisaccade performance for the PD patients was degraded for both the volition decision process and the initiation of saccade with direction opposite to the cue. Also, volition latency analysis is a more objective method than prosaccade and antisaccade latency analysis, which can be compared among results obtained from different analysis methodologies. PMID- 10672790 TI - A 16-channel SQUID-device for biomagnetic investigations of small objects. AB - Biomagnetic investigations in basic physiological research using animals require measurement devices different from commercial biomagnetometers used in human investigations. Two major problems have to be tackled in the design of such biomagnetometers. First, the spatial sampling needs to be much higher. Second, the distance between pick-up coils and the sources needs to be much shorter in order to compensate the worse signal-to-noise ratio (SNR) due to the smaller pick up coils. We designed and built a 16-channel biomagnetic measurement system meeting these design criteria. The pick-up coil diameter of this new biomagnetometer is 6.7 mm, thus allowing 16 channels on an area of 3.2x3.2 cm2. The pick-up coils are located 3 mm above the dewar outer bottom, hence the closest distance to the cortical surface can be a few millimetres. We provide as an example of first measurements performed with the new biomagnetometer investigations of epileptic spikes in adult rabbits by simultaneous magnetoencephalogram (MEG) and electrocorticogram (ECoG) recordings. The high SNR of the recorded MEG and the simultaneously detected electric potentials allow investigations of the spatio-temporal pattern of neuronal processes of epileptiform spikes with signal strengths of about 3.5 pT. PMID- 10672791 TI - The incidence of radio-frequency impulsive noise within hospital buildings: initial measurements in the 450 MHz band. AB - This paper reports on the incidence and level of man-made electrical impulse noise capable of causing interference to low-power, ASK and FSK radio biotelemetry links. Measurements were made at two acute hospitals, one in a residential area of Belfast, Northern Ireland and the other in an industrial part of Berlin, Germany. Monitoring was effected in the 450 MHz band using a communications receiver with its AGC line disabled and AM-detector AM output fed to a PC-based logger. The RF bandwidth was 20 kHz and a rotatable folded-dipole receiving antenna was used. Seven-day impulse counts were recorded at each location, for separate RF input thresholds of -100, -110 and -120 dBm and a common trigger window of 10 micros-10 ms. Histograms depicting the average pulse count per minute are presented; values range from a maximum of 55x10(3)/min for 120 dBm sensitivity in Berlin, to a minimum of 1.2/min for -100 dBm sensitivity in Belfast, both with vertical polarisation. PMID- 10672792 TI - Variation of the spatial position computed by Roentgen Stereophotogrammetric Analysis (RSA) under non-standard conditions. AB - Roentgen Stereophotogrammetric Analysis (RSA) has been applied to different kinds of research in order to obtain important information in the biomechanics field. Operative requirements change according to the type of investigation and sometimes practical conditions do not always permit one to respect standard specifications. The aim of this paper is to verify the reliability of the system under non-standard conditions, studying the effects of the focus-to-film distance on the determination of 3D co-ordinates of markers. The application of statistical analysis, consisting of two and three way ANalysis Of VAriance (ANOVA), showed that focus-to-film distance is not a significant source of variation, i.e., the system can correctly compute the marker position inside the calibrated space under all conditions of magnification. Since we obtained a fluctuation due to focal distance change of 10(-5) mm (LSD for 95% confidence level), we can assert that this source causes a variation strictly within the limits required by clinical investigations. Thus, this study contributes to improving the flexibility of RSA in clinical applications, making the technical requirements of the radiographic set-up less constraining. PMID- 10672793 TI - Calculation of extracellular potentials produced by an inclined muscle fibre at a rectangular plate electrode. AB - Generally the anatomy of muscles is rather complex, and the fibres have various inclination angles within the muscles. We suggest a fast and reliable way to calculate extracellular potentials produced at a point or rectangular plate electrode by a muscle fibre of finite length with an arbitrary inclination. A muscle fibre was considered to be a linear timeshift-invariant system of potential generation. Then, similar to the fibre without inclination, the extracellular potential produced by an inclined fibre was represented as the output signal of the system; it was calculated as the convolution of the input signal and impulse response. Irrespective of the inclination, the input signal of the system was the first temporal derivative of the intracellular action potential. The impulse response of the system differed for the fibres with inclination. This required a new method of analytical integration over the rectangular electrode area. The approach provides a chance to simulate and analyze motor unit potentials or F-, H- or M-responses produced by muscles of complicated anatomy (circum-pennate or complex pennate type) at electrodes of actual size and location in normals and patients with neuro-muscular disorders. PMID- 10672794 TI - Dimensional change in muscle as a control signal for powered upper limb prostheses: a pilot study. AB - The vast majority of externally powered prostheses are controlled from the myoelectric signal, measured at the skin surface using socket-located electrodes. This signal has been well researched and sophisticated signal processing methods developed. Nevertheless, the inherent properties of the signal, such as its broad bandwidth and low voltage amplitude, make its use less than straightforward in the control of low frequency activity such as powered prosthetic hand movement. This paper reports on a pilot study of an alternative, a signal derived from dimensional change in muscle. A new socket-located sensor was designed to measure dimensional change in muscle, the linearised output of which is termed the myokinemetric (MK) signal. This was used in a series of tasks aimed at investigating the potential for its use in upper-limb prosthesis control. Six amputee subjects were tested, of whom one was a regular user of the myoelectric hand, one had some experience, and four had little or no previous experience of controlling devices using their residual limb. Data is presented on the problems of shift in signal range with time and socket donning and doffing and on the ability of subjects to control the amplitude of the signal. The results show that subjects were able to control the magnitude of the MK signal to a significant degree, with typical errors averaging 0.1-0.3 mm, around 10% of the signal range. The principal problem encountered was the shift in signal with time and socket donning and doffing. PMID- 10672795 TI - Bone donor site. Iliac crest or distal radius? AB - In a series of 18 patients requiring bone-grafting procedures to the wrist, bone structure and bone turnover characteristics were compared between samples of bone harvested from the iliac crest and the distal radius. Histologically and biologically the iliac crest bone was found to be superior to distal radial bone. Axial bone was also shown to be more resistant to the ravages of increasing age and chronic disease. However, in spite of these laboratory findings, clinically, radial bone is equally successful as graft material. This, in all probability, is due to the greater porosity of radial bone which allows better compaction, more accurate filling of defects and easier nourishment of the graft by surrounding body fluids. PMID- 10672796 TI - Early active mobilization of primary repairs of the flexor pollicis longus tendon. AB - This study reports the treatment of divided flexor pollicis longus (FPL) tendons from 1989 to 1998. The first 30 patients, in whom the tendon was repaired with a Kessler suture and simple epitendinous suture and mobilized using early active motion with only the thumb splinted, achieved 70/73% (White/Buck-Gramcko assessments respectively), excellent or good results and a rupture rate of 17%. The next 39 patients underwent the same treatment but in a splint with the thumb position altered and the fingers also splinted, with 67/72% excellent or good results and a rupture rate of 15%. The next 49 patients underwent repair with a Kessler suture and a reinforced epitendinous suture and the same mobilization as group 2, with 76/80% excellent or good results and a rupture rate of 8%. The final combination of repair and early active mobilization for primary repair of FPL tendons compares favourably with previous methods of treatment. PMID- 10672797 TI - Flexor tendon repair using a stainless steel external splint. Biomechanical study on human cadaver flexor tendons. AB - A stainless steel external tendon splint was used in repair of cadaver tendons and compared with standard tendon repairs with suture. The splint was combined with a Kessler repair and tested against the Kessler, Becker, and Savage repairs in fresh human cadaver flexor digitorum profundus tendons. Biomechanical testing was done on a tensile testing machine, and load-displacement curves were generated. The repairs using the external tendon splint demonstrated a range of improvement of 32 to 146% in mean maximal tensile strength and a 20 to 185% improvement of mean ultimate tensile strength compared with all other repairs. The external tendon splint is relatively easy to apply to a tendon. The repair is strengthened and becomes capable of withstanding early active range of motion exercises. In vivo testing will be needed to assess the potential clinical usefulness of such a device. PMID- 10672798 TI - Cytotoxicity of cyanoacrylate adhesives to cultured tendon cells. AB - The in vitro cytotoxicity of four cyanoacrylate adhesives was tested using cultures of cells derived from human tendons. All four were found to be cytotoxic, even at concentrations as low as 1.7%, over the experimental period of up to 18 weeks. This study shows that such adhesives in their present state may not be suitable for re-joining cut tendons as their initial and long-term toxicity may hinder the slow healing process of tendons. PMID- 10672799 TI - Open and closed arthrodesis of the rheumatoid wrist using a modified (Stanley) Steinmann pin. AB - In a series of 21 patients (22 wrists) with rheumatoid arthritis, arthrodesis of the wrist was done using a modified Steinmann pin (Stanley) either by an open or closed technique. The open technique, which included fragmenting the carpal bones (12 cases), was mainly used when additional procedures were needed simultaneously. The closed technique simply required insertion of the Stanley pin under fluoroscan control through a small incision over the metacarpal head. Nine out of 12 wrists treated with the open technique and nine out of ten of those treated by the closed technique were successfully fused. Complications were few. A single patient was dissatisfied due to continuing pain. Two out of the four pins that migrated (both involving the open technique) have been removed. PMID- 10672800 TI - Comparative results of resection of the distal ulna in rheumatoid arthritis and post-traumatic conditions. AB - The purpose of this study was to determine whether the results of resection of the distal ulna differed depending upon the underlying aetiology of the condition. Patients with rheumatoid arthritis were compared with patients with post-traumatic wrist complaints. Fifty resections in 40 patients (eight male, 32 female) were assessed with respect to pain, range of motion, and grip strength. Of the 23 rheumatoid wrists, 86% were pain-free following surgery; however, only 36% of the patients in the trauma group reported pain relief postoperatively. Pain relief in post-traumatic patients was more predictable when distal radioulnar joint arthrosis was identified as the sole cause of wrist pain. PMID- 10672801 TI - Results of the wafer procedure for ulnar impaction syndrome in the ulnar negative and neutral wrist. AB - Thirteen wrists with ulnar neutral or negative variance were treated by open distal ulna excision (the wafer procedure). The mean follow-up was 25 months (range, 12-38). At final follow-up grip strength had increased a mean of 14 kgf and 12 of the 13 patients were very satisfied with the functional outcome and pain relief. In treatment of the ulnar impaction syndrome, the wafer procedure provides excellent pain relief and functional restoration particularly in patients with ulnar neutral or negative wrists in whom triangular fibrocartilage tears have not yet developed. PMID- 10672802 TI - Internal fixation of scaphoid fractures with an AO mini-fragment lag screw, using temporary interoperative AO mini external fixation. PMID- 10672803 TI - Repair of central slip avulsions using Mitek Micro Arc bone anchors. An in vitro biomechanical assessment. AB - In this study we evaluated the pullout strength of the Mitek Micro Arc anchor for the reconstruction of central slip avulsions at the proximal interphalangeal joint of the finger. Forty paired fresh frozen cadaver fingers were randomized into treatment (anchor) and control groups (horizontal mattress repair) and subjected to tensile loading to failure. The mean (SD) failure loads of the repairs were: Mitek repair group 22.3 (4.7) N, and control group 24.7 (5.5) N. There were no statistically significant differences between the failure loads or the failure mechanisms of the two repairs. The pullout strength of the isolated anchor-bone complex was evaluated by refitting five anchors with stainless steel wire. The mean failure load of the isolated anchor was 400% higher than the tendon-suture-anchor complex, indicating that the weakest link of the system is not the bone-anchor interface. PMID- 10672804 TI - Fixation of metacarpal fractures using absorbable hemi-cerclage sutures. AB - We retrospectively reviewed the use of biodegradable hemi-cerclage sutures in the treatment of 79 metacarpal fractures in 66 patients. The polyglycolic acid hemi cerclages achieved sufficient fracture fixation to permit early motion exercises, but fractures were also immobilized for a mean of 3.7 (range, 1.5-6) weeks postoperatively, during which time physiotherapy was given. Adequate bony stability was achieved after a mean of 4.5 (range, 3.5-7) weeks and fracture redisplacement occurred in only one case. PMID- 10672805 TI - Internal suture for mallet finger fracture. AB - An internal suture technique has been used for mallet finger fractures involving at least 30% of the articular surface. It provides fixation without a button or transfixion of the fragment. An independent retrospective review was conducted of ten patients at a mean follow-up of 17 months. Mean visual analogue score (0 to 10) for pain was 2.4 and satisfaction 7.9. Mean active range of motion was 13 to 49 degrees, passive motion was 2 to 56 degrees, pinch strength of effected finger to thumb was 3.8 kgf (81% of the opposite finger), grip strength 37.9 kgf (95% of the opposite hand). All fractures united and there were no neuromas. Complications included two nail deformities, a superficial infection and a pin track infection. One patient with a crush injury continued to have pain despite an arthrodesis. PMID- 10672806 TI - Nonunion following subcapital (neck) fractures of the proximal phalanx of the thumb in children. AB - Six cases of nonunion of subcapital (neck) fractures of the proximal phalanx of the thumb in children were seen over a period of 5 years. Ages at the time of injury ranged between 2 and 3 years. Entrapment of the thumb in a closing door was the mechanism of injury in all cases. All fractures were closed and were significantly displaced. Immediate management was by closed reduction and splinting in four cases, closed reduction and K-wire fixation in one case and no treatment in one case, which was later treated by delayed open reduction and K wire fixation. Only two of the six ununited fractures were eventually treated with bone grafts and both fractures united resulting in a stable thumb but with a limited range of flexion of the interphalangeal joint. Factors that may increase the risk of nonunion of these fractures in children are discussed. PMID- 10672807 TI - Simultaneous fracture-dislocations of the distal and proximal interphalangeal joints. AB - Sixteen cases of simultaneous fracture-dislocations of both the distal interphalangeal (DIP) and proximal interphalangeal (PIP) joints in the same finger that were treated during the past 10 years were classified into three types: the swan-neck injury (dorsal fragment of the base of the distal phalanx at the DIP joint and palmar fragment of the base of the middle phalanx at the PIP joint); the double-hyperextension injury (palmar fragments at the DIP and PIP joints); and the straight-finger injury (with dorsal and palmar bone fragments at the DIP joint). The results of treatment were more satisfactory in PIP joints than in DIP joints. PMID- 10672808 TI - The use of silicone tubing in the late repair of the median and ulnar nerves in the forearm. AB - A silicone tube segment was used for repairing the median and ulnar nerves in the forearm. This study includes 26 patients (20 male and six female), with a mean age of 23 years (range, 18-26). Injuries were caused by saw, knife and glass accidents, the latter being most frequent. The mean interval between the injury and repair was 101 days. Fourteen patients had median nerve injuries, eight had ulnar nerve injuries and four had both median and ulnar nerve injuries. The technique was effective in the repair of peripheral nerve injuries with gaps of up to 3 cm, with better results in the ulnar nerves than in the median nerves. PMID- 10672809 TI - The influence of delay and the effect of fibrin sealant on the cut surface of the peripheral nerve. An experimental study in the rat. AB - Thirty-two sciatic nerves in 16 rats were divided to investigate the effect of delay in fixation and the use of fibrin sealant at the site of the division on the nerve end. Specimens were assessed by morphological and morphometric criteria using scanning electron microscopy and longitudinal sections. All specimens showed a protruded nerve end. Wrapping the nerve with fibrin sealant before division and immediate fixation of the specimen resulted in less protrusion. PMID- 10672810 TI - Psychological aspects of toe to hand transfer in children. Comparison of views of children and their parents. AB - Thirty-seven children with congenital (n = 32) or post-traumatic (n = 5) hand anomalies underwent unilateral or bilateral toe transfers. All had undergone preoperative counselling. After rehabilitation and more than 1 year after surgery, the children and their parents were reviewed by a clinical psychologist to assess the psychosocial outcome of the surgery. A high level of satisfaction was reported with regard to the surgery, in terms of function, cosmesis, donor site, psychosocial wellbeing and the reactions of others. This was true regardless of the gender of the child. However, there was a tendency for the children to be more positive in their responses than their parents. PMID- 10672811 TI - 4.5 year follow-up after surgical correction of upper extremity deformities in spastic cerebral palsy. AB - Reconstructive surgery was carried out on 27 upper extremities in 24 children with deformities due to spastic cerebral palsy. Functional evaluation of the affected extremities was made preoperatively, at 6 months and at a mean of 4.5 years postoperatively using a score added to the assessment system described by the Committee on Spastic Hand Evaluation. According to the score, dysfunction of the arm was significantly reduced 6 months after the reconstructive surgery and the improvements remained essentially unchanged at the later follow-up. The addition of a score to the original assessment system facilitated the overall assessment of postoperative results. PMID- 10672812 TI - Optimal position for the one-bone forearm. An analysis using a hinged brace in normal subjects. AB - The purpose of the present study was to determine whether there is an optimal position for fixation of the one-bone forearm. Eight normal individuals were fitted with a hinged brace which fixed the position of pronation and supination and underwent functional hand testing using the tests of Jebsen et al. Our results indicate that a one-bone forearm in a position of 30 degrees of pronation will provide the best function for writing and working with small objects using the dominant arm. PMID- 10672813 TI - The role of Mepitel silicone net dressings in the management of fingertip injuries in children. AB - Forty-five children with isolated fingertip injuries were randomized for treatment with either Mepitel silicone net dressings or paraffin gauze dressings. Over a 4 week period, the objective adherence of the dressing, and the perceived level of stress caused to the child by the dressing change were scored by linear analogue scales. The wounds were also assessed for the progress of healing and presence of infection. Twenty children received Mepitel dressings and 25 had paraffin gauze dressings. There was no difference in duration of healing or complication rates between the two groups. Statistically lower scores were seen for the Mepitel group for the first 3 weeks in both adherence and stress scores. These results suggest that silicone net dressings may be a less adherent and less painful method of dressing fingertip injuries in children. PMID- 10672815 TI - Congenital aneurysm of the ulnar artery in the palm. AB - An aneurysm of the palmar ulnar artery is rare and usually follows penetrating or blunt trauma to the hypothenar eminence. We report a case in which there was no history of trauma and the aneurysm may have been of congenital origin. PMID- 10672814 TI - Hypothenar hammer syndrome. Retrospective study of nine cases. AB - The hypothenar hammer syndrome is an uncommon lesion of the ulnar artery caused by repetitive trauma to the ulnar portion of the hand. It characteristically occurs in the dominant hand of middle-aged craftsmen, but also in athletes practising various types of sports. We present a retrospective study of nine patients between 1988 and 1999. The follow-up ranged from 1 to 10 years. We recommend surgical treatment, by resection of the involved arterial segment and revascularization either by direct anastomosis or by means of a venous interpositional graft. PMID- 10672816 TI - Sympathetic block significantly improves reperfusion in skeletal muscle following prolonged use of tourniquet. AB - The effects of guanethidine sympathetic nerve blocks on reperfusion of skeletal muscle was studied in rats. After 3 hours of ischaemia reperfusion was significantly better in animals that had received guanethidine. PMID- 10672817 TI - Delayed sequential ruptures of the index and thumb flexor tendons due to an occult scaphoid nonunion. AB - A case of sequential spontaneous loss of flexion in the thumb and index finger over a 10 year period is reported. Previously unrecognized carpal collapse as a consequence of a longstanding scaphoid nonunion was identified as the cause. Fusion of the interphalangeal joint of the thumb and suture of the index profundus tendon to the middle profundus tendon resulted in a good outcome. PMID- 10672818 TI - Proximal row transcarpal fracture from a punching injury. AB - We describe an unusual case of a 31-year-old woman who injured the right dominant wrist when she punched an assailant's shoulder. She described a mechanism of direct compression, with the wrist in hyperextension, radial deviation and the forearm in pronation. She sustained an oblique transverse fracture of the proximal pole of the scaphoid and a coronal plane fracture of the lunate and the triquetrum. This unusual proximal row transcarpal fracture is in conflict with the Mayfield sequence and was caused by a low velocity injury. PMID- 10672819 TI - Periosteal chondroma of the distal radius. AB - We report a case of periosteal chondroma in an unusual location at the distal end of the radius. PMID- 10672820 TI - Unusual complication of ligation of rudimentary ulnar digit. AB - We report a case of rudimentary ulnar polydactyly of the hand of a 7-year-old female child. Histological examination revealed a central traumatic neuroma which branched into five digit-like projections covered with hyperkeratotic epidermis. We think this was a result of suture ligation during the postnatal period. PMID- 10672821 TI - Dupuytren's disease in Marfan's syndrome. AB - We report a case of histologically confirmed Dupuytren's disease of the hand in a young man with Marfan's syndrome. PMID- 10672822 TI - Procedures on the distal end of the ulna are given the name of the surgeon or surgeons who described them. PMID- 10672824 TI - The Mantero technique is inappropriate because of a risk of infection due to external material and burying it can overcome that problem PMID- 10672823 TI - The Mantero flexor tendon repair in zone 1. PMID- 10672825 TI - An aid to digit replantation: a new use for the Harris silicone tile. PMID- 10672826 TI - Recurrent osteoblastoma of the hamate bone. PMID- 10672827 TI - Classical Barrett esophagus contrasted with Barrett-type epithelium at normal appearing esophagogastric junction. Central Finland Endoscopy Study Group. AB - BACKGROUND: Incomplete intestinal metaplasia or specialized columnar epithelium (SCE) is the histologic hallmark of Barrett esophagus (BE), but it may also occur at a normal-appearing gastroesophageal junction without BE. We studied whether differences occur between BE patients and those with SCE at the squamocolumnar junction but without BE (abbreviated JSCE), in terms of endoscopic and histologic signs of gastroesophageal reflux disease (GERD) and Helicobacter pylori gastritis. METHODS: A total of 1059 consecutive patients referred for endoscopy in one hospital district in Finland were enrolled in the study. Biopsy specimens (at least two from each site) were obtained from the gastric antrum and the corpus of the stomach and from the esophagogastric junction and distal esophagus. RESULTS: Classical BE was detected in 25 (2%) and JSCE in 99 (9%) patients. Dysplasia in the metaplastic mucosa was observed in six BE patients but in none of the JSCE patients (P < 0.001). In multivariate analysis the independent risk factors for BE were endoscopic erosive esophagitis (odds ratio (OR), 6.08; 95% confidence interval (CI), 2.50-14.82), male sex (OR, 3.02; 95% CI, 1.20-7.65), and age (OR, 1.02 per year; 95% CI, 1.00-1.06). The independent risk factors for JSCE were endoscopic erosive esophagitis (OR, 1.88; 95% CI, 1.08-3.29) and age (OR, 1.03; 95% CI, 1.02-1.05) but not H. pylori infection (OR, 1.57; 95% CI, 0.83 2.97) or chronic gastritis (OR, 0.88; 95% CI, 0.44-1.75). In univariate analysis, however, JSCE was associated with antral-predominant atrophic gastritis (77% H. pylori-positive). Unlike in JSCE patients, male sex strongly predominated among BE patients (P = 0.01). The mean ages of BE and JSCE patients did not differ. CONCLUSIONS: Both BE and JSCE without BE increase in prevalence with age, and both associate with endoscopic erosive esophagitis but not with H. pylori gastritis. However, because of the marked sex disparity, JSCE cannot be a direct precursor of BE, and some factors other than GERD alone also play a role in the pathogenesis of BE. Compared with BE, dysplasia is a rare finding in JSCE, and endoscopic surveillance with biopsy specimens from JSCE patients without dysplasia is not recommended. PMID- 10672828 TI - Screening for oesophageal adenocarcinoma: an evaluation of a surveillance program for columnar metaplasia of the oesophagus. AB - BACKGROUND: Screening patients with columnar metaplasia of the oesophagus for adenocarcinoma is controversial owing to the low cancer incidence and diverging opinions as to whether screening improves the prognosis of these patients. Our aim was to evaluate a screening program for adenocarcinoma in patients with columnar metaplasia in the oesophagus, with focus on cancer incidence and costs. METHODS: One hundred and ninety-nine patients with columnar metaplasia of the oesophagus were identified through an endoscopy database, and the original patient records were reviewed. RESULTS: The patients were followed up for 797 years in total and during this time were subjected to 1071 upper gastrointestinal endoscopies. During the screening period 5 patients presented with adenocarcinoma; thus the cancer-incidence was 1 in 159 patient-years. The cost of detecting one cancer was 294,950 SEK (US$ 37,815). However, only four of the five patients were suitable for oesophagectomy, and of these, one patient turned out to have an advanced cancer. All patients developing cancer had columnar metaplasia of the oesophagus longer than 3 cm and specialized columnar epithelium (intestinal metaplasia/Barrett oesophagus). CONCLUSIONS: Low cancer incidence, high costs, and the doubtful prognosis for the patients with identified cancer question the benefits and cost-effectiveness of cancer screening among patients with columnar metaplasia in the oesophagus. PMID- 10672829 TI - The esophageal Z-line appearance correlates to the prevalence of intestinal metaplasia. AB - BACKGROUND: Intestinal metaplasia at the gastroesophageal junction is associated with Barrett esophagus, gastric cardiac cancer, and gastritis. The aim of this study was to determine the prevalence of intestinal metaplasia among patients with symptoms suggestive of gastroesophageal reflux disease (GERD) and to study clinical, endoscopic, and histologic associations with intestinal metaplasia at the squamocolumnar junction. METHODS: One hundred and eighty-six patients with symptoms indicating gastroesophageal reflux were included in the study. A new classification of the Z-line appearance was used. RESULTS: The Z-line appearance was found to correlate with the prevalence of intestinal metaplasia at the squamocolumnar junction (P = 0.0001). Intestinal metaplasia at the squamocolumnar junction was found in 15.0% of the patients. There was a statistically significant association between intestinal metaplasia at the squamocolumnar junction and tongues of columnar epithelium at the Z-line (P = 0.020), intestinal metaplasia in the cardia (P = 0.020), positive CLO test (P = 0.026), smoking (P = 0.041), and age (P = 0.050). There was no association with endoscopic or histologic signs of esophagitis or with the severity or duration of GERD symptoms. CONCLUSION: Intestinal metaplasia at the squamocolumnar junction correlates with the Z-line appearance, which would justify a new classification. PMID- 10672830 TI - Expression of hepatocyte growth factor and c-met receptor in gastric mucosa during gastric ulcer healing. AB - BACKGROUND: Recent studies have shown that hepatocyte growth factor (HGF) has a mitogenic and motogenic activity for gastric mucosal cells. The aim of this study is to ascertain the clinical significance of the expression of HGF and its receptor (c-met) during the course of gastric ulcer healing. METHODS: Ten patients who underwent endoscopic mucosal resection (EMR) and developed acute gastric ulcer and 20 patients with peptic ulcer of the stomach were enrolled in this study. Tissue samples were serially obtained from both the edge of the ulcer and its neighboring mucosa 0, 2, and 8 weeks after EMR or diagnosis. Gene expression of HGF, c-met, c-myc, and proliferating cell nuclear antigen (PCNA) was semiquantitatively examined by reverse transcription polymerase chain reaction (RT-PCR) using beta-actin as an internal control. In addition, expression of HGF and c-met mRNA with regard to the presence or absence of Helicobacter pylori infection was studied. RESULTS: In patients after EMR HGF, c met, c-myc, and PCNA mRNA in the ulcer lesion at 0 week was at the peak of its expression and then decreased with time (P < 0.01, P < 0.05, P < 0.05, and P < 0.05, respectively). In patients who underwent EMR the intensity of HGF mRNA was correlated with that of c-met, c-myc, and PCNA mRNA (P < 0.0001, P = 0.0095, and P = 0.0009, respectively) and that of c-met mRNA was correlated with that of c myc and PCNA mRNA (P = 0.012 and P < 0.0001, respectively). In patients with peptic ulcer, expression of HGF mRNA decreased with time (0 week versus 8 weeks, P < 0.05; 0 week versus 8 weeks, P < 0.01). c-Met expression was lower at 8 weeks than at 0 week (P < 0.05). The intensity of c-met mRNA was correlated with that of PCNA mRNA (P < 0.0001). H. pyiori-infected mucosa had a higher level of HGF mRNA than non-infected mucosa. CONCLUSION: The present study suggests that HGF is one of the growth factors acting together during healing of gastric ulcer. PMID- 10672831 TI - Alpha1-antitrypsin deficiency may be a risk factor for duodenal ulcer in patients with Helicobacter pylori infection. AB - BACKGROUND: Most individuals with Helicobacter pylori infection in Western countries have no evidence of peptic ulcer disease (PUD). We therefore assessed the PiZ deficiency variant of the major plasma protease inhibitor alpha1 antitrypsin (alpha1AT) as a risk factor for PUD in H. pylori-infected individuals. METHODS: The cohort comprised 100 patients with endoscopically or surgically proven PUD (30 patients with duodenal ulcer (DU) and 70 patients with gastric ulcer (GU)) and 162 age- and sex-matched controls with PUD-negative endoscopic findings and no history of PUD. Plasma samples were screened for alpha1AT deficiency (PiZ) with an enzyme-linked immunosorbent assay (ELISA) and phenotyped by isoelectric focusing. H. pylori infection was evaluated with an IgG ELISA technique. RESULTS: Among the 262 patients 17 (6.5%) were positive for the PiZ alpha1AT deficiency, a frequency of the same magnitude as in the Swedish general population (4.7%). Of the PiZ carriers 76% (13 of 17) had H. pylori antibodies compared with 61% (151 of 245) of the non-PiZ carriers (NS). The prevalence of DU tended to be higher in H. pylori-positive PiZ carriers than in non-PiZ carriers (15.4%, 4 of 26 versus 0 of 4). Furthermore, among patients with DU a high PiZ allele frequency (13.3%, 4 of 30) was found compared with the general population (4.7%) (odds ratio (OR), 3.2; 95% confidence interval (CI), 1.09-8.94; P = 0.02). All DU patients carrying the PiZ allele were positive for H. pylori. In addition, four of five PiZ carriers with H. pylori infection and PUD had DU. CONCLUSIONS: The PiZ allele may be a contributing factor in the development of DU in H. pylori-positive individuals. PMID- 10672832 TI - Evaluation of the clinical relevance of the iceA1 gene in patients with Helicobacter pylori infection in Japan. AB - BACKGROUND: A novel Helicobacter pylori gene, iceA, has two allelic variants, and the iceA1 strain is associated with peptic ulcer disease. The aims of this study were to evaluate whether the possession of iceA1 gene is associated with gastric cancer or the severity of gastritis. METHODS: Ninety-seven subjects (46 patients with early gastric cancer and 51 control subjects) infected with H. pylori were studied. DNA was extracted from isolated H. pylori strains, and the presence of the iceA1 gene was examined by polymerase chain reaction. The features of gastritis were graded in accordance with the updated Sydney System, using gastric biopsy specimens. RESULTS: iceA1 was found in 61% of patients with gastric cancer and 53% of control subjects (NS). The grade of gastritis in iceA1-positive and negative gastric mucosa was compared. Higher polymorphonuclear cell infiltration was observed in iceA1-positive subjects (P < 0.05). However, no significant difference was observed in the grades of mononuclear cell infiltration, glandular atrophy, and H. pylori density. CONCLUSIONS: Our results suggest that the iceA1 gene is not associated with the development of gastric cancer in Japan, whereas the iceA1-positive strain may induce more enhanced active gastric inflammation in cagA-positive and vacA s1/m1 strains. PMID- 10672833 TI - Apoptosis of human gastric epithelial cells via caspase-3 activation in response to Helicobacter pylori infection: possible involvement of neutrophils through tumor necrosis factor alpha and soluble Fas ligands. AB - BACKGROUND: Infection with Helicobacter pylori activates a proinflammatory gene program in human gastric epithelial cells and neutrophils and is associated with significant epithelial cell damage, including an increased level of apoptosis. We evaluated whether immune mediators produced by neutrophils could modulate gastric epithelial cell apoptosis in response to H. pylori infection. METHODS: After gastric epithelial cells were infected with H. pylori in the presence of immune mediators, including tumor necrosis factor alpha (TNF-alpha) and Fas ligand (FasL), apoptosis and caspase-3 activity were assessed. The neutrophils were obtained from healthy volunteers, and Western blot for FasL and quantitative reverse transcription polymerase chain reaction for TNF-alpha transcripts were performed. Fas expression in gastric epithelial cells was explored by flow cytometric analysis. RESULTS: Activation of caspase-3 was first apparent 12 h after bacterial infection, and the phenotypic expression of apoptosis was first apparent 18 h after bacterial infection. The extent of apoptosis was similar in cases of cagA+ cytotoxin+, cagA+ cytotoxin-, cagA- cytotoxin- H. pylori-infected gastric epithelial cells. Approximately 20% of the Hs746T cells expressed Fas within 24 h of H. pylori infection. The soluble FasL was upregulated in neutrophils after treatment with H. pylori-soluble proteins for 24 and 48 h. The addition of TNF-alpha and the soluble form of FasL, produced by neutrophils, significantly increased H. pylori-infected cell apoptosis and caspase-3 activation. However, the combination of these two immune mediators showed only an additive increase. CONCLUSION: These results suggest that H. pylori-induced gastric epithelial cell apoptosis and caspase-3 activation can be modulated by the immune response of neutrophils. PMID- 10672834 TI - Usefulness of a novel enzyme immunoassay for the detection of Helicobacter pylori in feces. AB - BACKGROUND: In this study we assessed the reliability of a newly developed enzyme immunoassay (HpSA) kit for detecting Helicobacter pylori antigen in stool. METHODS: This study included 309 patients, 147 of whom were defined as positive and 162 as negative by the 13C-urea breath test, rapid urease test, and pathologic findings. From these patients fresh stool specimens were collected for HpSA. RESULTS: When 0.100 was adopted as the cut-off value, in accordance with the manufacturer's recommendations, the sensitivity, specificity, and accuracy of the HpSA were 98.0%, 87.0%, and 92.2%, respectively. However, these values were much improved when a cut-off value of 0.300 was adopted, which was obtained with our receiver-operator characteristics curve; with this value the sensitivity, specificity, and accuracy of HpSA were 93.9%, 95.7%, and 94.8%, respectively. CONCLUSION: These results indicate that HpSA is a highly reliable diagnostic method for H. pylori infection and is useful in confirming eradication. PMID- 10672835 TI - Screening of patients with acute infectious diarrhoea: evaluation of clinical features, faecal microscopy, and faecal occult blood testing. AB - BACKGROUND: For optimal management of acute infectious diarrhoeal diseases, it is necessary to utilize a screening process to distinguish between invasive and non invasive diarrhoeas. The aim of this study was to compare the diagnostic utilities of clinical features, faecal microscopy (FM), and faecal occult blood testing (FOBT) in distinguishing invasive diarrhoeas from non-invasive ones. METHODS: A total of 1008 patients with acute diarrhoea were evaluated. Rectal swabs were cultured for Salmonella, Shigella, and Vibrio species; rectal swabs from 109 of these patients were also examined for Campylobacter, enterotoxigenic Escherichia coli, and rotavirus species. Isolation of faecal enteropathogens served as the gold standard. FOBT was performed with a commercial modified guaiac test. Specificity, sensitivity, positive predictive value (PPV), negative predictive value (NPV), accuracy, and likelihood ratio were compared. RESULTS: Among the 1008 patients 402 with a single identified enteropathogen were available for analysis. Invasive and non-invasive enteropathogens were isolated from 262 (65.2%) and 140 (34.8%) cases, respectively. The presence of visible blood in faeces was almost a pathognomonic sign of invasive diarrhoea but had poor sensitivity. Clinical features were useful but inadequate in differentiating patients with non-bloody diarrhoea (74% of patients) into invasive and non invasive categories. The sensitivities, specificities, PPVs, and NPVs of FM and FOBT were 75%, 77%, 58%, 88%, and 85%, 68%, 53%, and 91%, respectively. CONCLUSION: The presence of visible blood in faeces is a highly specific clinical feature of invasive diarrhoea but suffers from low sensitivity. In non-bloody diarrhoea FOBT is a valuable screening test and is comparable to FM, particularly when interpreted in the clinical context. PMID- 10672836 TI - No polymorphism in the tissue transglutaminase gene detected in coeliac disease patients. AB - BACKGROUND: The autoantigen for the anti-endomysial antibody (AEA) found in coeliac disease has recently been reported to be the enzyme tissue transglutaminase (tTG). Polymorphisms in the gene for tTG would result in different enzymic isoforms being expressed. Certain isoforms may interact with gliadin to create antigenic neoepitopes, which could then generate an immune response in genetically predisposed individuals possessing major histocompatibility complex (MHC) class II DQ2. METHODS: We have sequenced the coding region of tTG in coeliac patients and normal controls. Total RNA was extracted from mucosal biopsies from eight AEA-positive histologically proven coeliac disease patients and four control patients with a histologically normal duodenum and a negative AEA. The 2-kb coding region of tTG was amplified in overlapping fragments by reverse transcription polymerase chain reaction (PCR), using five sets of PCR primers. Each overlapping PCR fragment was sequenced using the fmol DNA sequencing system. RESULTS: tTG transcripts were detected in all samples. There was no difference in the coding sequence of tTG between the four control and eight coeliac patients, even though we observed differences in sequence between our study and the original published sequence. These differences have also been reported in sequences published subsequent to the original description. CONCLUSIONS: Polymorphisms in the tTG gene have not been observed in coeliac disease patients and therefore cannot explain the creation of neoepitopes. PMID- 10672837 TI - Relationship between intestinal permeability and calprotectin concentration in gut lavage fluid. AB - BACKGROUND: Calprotectin is released from neutrophils and monocytes, and increased calprotectin levels in stool may serve as a marker of intestinal inflammation. Intestinal permeability is increased in inflammatory bowel diseases, especially in Crohn disease. We studied the relationship between intestinal permeability and calprotectin concentration in intestinal lavage fluid in patients with known or suspected inflammatory bowel disease (IBD). METHODS: Thirty-eight patients were examined; 17 had Crohn disease; 3, ulcerative colitis; and 18, irritable bowel syndrome. Intestinal lavage was performed by means of a nasojejunal tube positioned by gastroduodenoscopy. By means of a peristaltic pump 2 l isotonic polyethylene glycol solution (MW, 3350) containing 50 microCi 51Cr labelled ethylenediaminetetraacetic acid (EDTA) were administered through the tube over a period of 40 min. The first clear fluid passed per rectum was collected and analysed for calprotectin levels with an enzyme-linked immunosorbent assay method. Urine was collected for 5 h and analysed for gamma radioactivity. 51Cr-EDTA excretion in urine was expressed as percentage of dose administered (that is, intestinal permeability). RESULTS: Both intestinal permeability and calprotectin concentration were significantly higher in patients with IBD than in patients with functional conditions. In Crohn disease the values depended on disease activity but not on whether the disease was located in the small or in the large bowel. There was a highly significant correlation between calprotectin concentration in gut lavage fluid and intestinal permeability (r=0.79, P<0.0001). CONCLUSION: The significant correlation between calprotectin concentration in gut lavage fluid and intestinal permeability supports the view that increased intestinal permeability in IBD might, at least in part, be a consequence of increased transepithelial migration of neutrophils. PMID- 10672838 TI - Abdominal symptoms: do they predict gallstones? A systematic review. AB - BACKGROUND: Our objective was to evaluate the diagnostic accuracy of abdominal symptoms in gallstones in studies using ultrasonography or oral cholecystography as the reference standard and to assess the extent to which variability in diagnostic accuracy is explained by patient selection and other characteristics of study design. METHODS: A Medline search (1966-1998) was conducted in combination with reference checking for further relevant publications. Two independent assessors selected controlled studies that included patients > or =18 years of age. Articles were excluded if sensitivity and specificity could not be extracted or the included patients were at extraordinary risk for gallstones. Seven abdominal symptoms were evaluated. Modification of the diagnostic accuracy by clinical setting, extent of the disease, blinding, age, and sex was analysed by using logistic regression. RESULTS: A total of 24 publications were included. The symptoms 'biliary colic', 'radiating pain', and 'analgesics used' were consistently related to gallstones. The setting of the study had a significant effect on the diagnostic accuracy of these symptoms. The unadjusted, pooled diagnostic odds ratios, however, were low (2.6 (95% confidence interval, 2.4 2.9), 2.8 (2.2-3.7), and 2 (1.6-2.5), respectively). The diagnostic odds ratio of biliary colic increased with the extent of gallstone disease (13.3 (4.2-42). CONCLUSIONS: Although biliary colic was specific for gallstones, 80% of the referred patients with gallstones presented with other abdominal symptoms. There is no current evidence that justifies the use of single abdominal symptoms, other than biliary colic, in the diagnosis of symptomatic gallstones. Further research should focus on the prognosis of patients with non-specific abdominal symptoms and gallstones. PMID- 10672839 TI - Preoperative diagnosis of bile duct strictures--comparison of intraductal ultrasonography with conventional endosonography. AB - BACKGROUND: The accuracy of intraductal ultrasonography (IDUS) and endoscopic ultrasonography (EUS) were compared in diagnosing biliary obstruction and in predicting surgical resectability. METHODS: Fifty-six patients with biliary obstruction were investigated preoperatively with both conventional EUS and IDUS. The ultrasonographic miniprobe was inserted into the bile duct system through the working channel of the duodenoscope during endoscopic retrograde cholangiopancreatography (ERCP). Conventional endosonography was performed with echoendoscopes in a standard technique. Images of endoluminal ultrasonography were prospectively reviewed and compared with intraoperative findings and resection specimen analyses. RESULTS: IDUS exceeded EUS in terms of accuracy (IDUS, 89.1%; EUS, 75.6%; P < 0.002), sensitivity (IDUS, 91.1%; EUS, 75.7%; P < 0.002), specificity (IDUS, 80%; EUS, 75%; NS), and T-staging (IDUS, 77.7%; EUS, 54.1%; P < 0.001). In bile duct carcinomas the accuracy rate for lymph node staging using IDUS (60%) is comparable with that using EUS (62.5%). In pancreatic carcinomas, however, lymph node staging using IDUS (13.3%) is significantly (P < 0.002) inferior to EUS (69.2%). Endoluminal ultrasonography may predict the potential resectability of bile duct tumors (IDUS, 81.8%; EUS, 75.6%; P < 0.002). CONCLUSIONS: IDUS proved to be accurate in preoperative diagnosing and T-staging of malignant biliary strictures, whereas it is not suitable for lymph node staging. IDUS using miniprobes during ERCP exceeds conventional EUS in terms of depiction of bile duct obstruction, diagnostic accuracy, and sensitivity and in the prediction of surgical tumor resectability. Additionally, different to EUS, IDUS can conveniently be performed during ERCP in one and the same session. PMID- 10672840 TI - Diet restriction increases ubiquinone contents and inhibits progression of hepatocellular carcinoma in the rat. AB - BACKGROUND: The aim of the present study was to evaluate the effect of a moderate diet restriction on the progression of preneoplastic foci into hepatocellular carcinomas (HCCs) and whether such an effect was related to altered cell proliferation, apoptosis, and/or tumour contents of lipid-soluble antioxidants. METHODS: Male Wistar rats were exposed to diethylnitrosamine as initiator and 2 acetylaminofluorene plus partial hepatectomy as promoter. Six weeks after initiation the animals were given a diet restricted to 75%-80% of that given to controls until being killed 45 weeks later. Macroscopic liver tumours were histologically classified. In hepatocellular carcinomas the numbers of S-phase (labelling index) and DNA-fragmented (apoptotic index) nuclei were calculated immunohistochemically, and the tumour contents of alpha-tocopherol and ubiquinone were determined. RESULTS: The number of animals with HCC and the number of HCCs per animal were significantly reduced in restricted-diet animals compared with controls. In HCCs the contents of ubiquinone-9 and -10 were significantly increased, labelling indices were enhanced 3-fold, and apoptotic indices 12-fold as a response to food restriction. Neither the size nor the differentiation of HCCs was altered by food restriction. The numbers and areas of preneoplastic foci were similar in restricted-diet animals compared with those of controls. CONCLUSION: Moderate, long-term food restriction inhibits the progression of preneoplastic liver foci into HCC. Possible mechanisms of this inhibition are a shift in the balance between apoptosis and cell division towards cell death and an adaptive response to oxidative stress by increased tumour contents of ubiquinones. PMID- 10672841 TI - C-met protooncogene expression and its regulation by cytokines in the regenerating pancreas and in pancreatic cancer cells. AB - BACKGROUND: Activation of the receptor c-met stimulates motility, mitosis, morphogenesis, processes involved in organ regeneration, or progression of malignancies. In the present study we investigated the expression of c-met protein in the regenerating pancreas and characterized the influence of cytokines on c-met expression. METHODS: Acute pancreatitis was induced in rats by cerulein injection. Rat acini and rat and human pancreatic cancer cells were stimulated with interleukin-1alpha (IL-1alpha), IL-6, tumor necrosis factor-alpha (TNF alpha) or transforming growth factor-beta1 (TGF-beta1). C-met expression was analyzed by means of Western blotting and localization in pancreatic tissue by immunohistochemistry. RESULTS: C-met protein expression was significantly upregulated in the regenerating pancreas and localized in areas of regenerating tissue. Stimulation with cytokines resulted in a two- to threefold increase of c met expression in vitro. CONCLUSION: Enhanced c-met expression after acute pancreatitis suggests that HGF/met has an important role in pancreatic regeneration, which is probably mediated by cytokines. This regulatory mechanism is also of importance in pancreatic cancer. PMID- 10672843 TI - Ultrasound-guided biopsies of abdominal organs with an automatic biopsy system. A retrospective analysis of the quality of biopsies and of hemorrhagic complications. AB - BACKGROUND: Ultrasound-guided biopsies of abdominal organs are not without risks for the patients; in particular, hemorrhagic complications may occur. Thus, over the last few years, automatic biopsy guns have been developed to facilitate the biopsy process. METHODS: The aim of our retrospective study was to examine the quality of specimens and the complication rate of ultrasound-guided biopsies of abdominal organs carried out in our institution using the automatic Autovac biopsy system during a period of 1.5 years. Of the total number of 321 biopsies, 290 were performed with the 1.2-mm Autovac needle, and in 31 cases the 0.95-mm needle was used. Among the 321 biopsies there were 211 of the liver parenchyma (66%), 47 of a liver tumor (14%), 38 of the pancreas (12%), 15 of the kidney parenchyma (5%), and 10 of a retroperitoneal tumor (3%). RESULTS: In 310 of the 321 biopsies it was possible to obtain sufficient diagnostically usable material for the pathologist (96.6%). In the other 11 cases the material obtained did not enable proper histologic diagnosis (3.4%). Two of these 11 biopsies were carried out with the 0.95-mm needle, and the other 9 with the 1.2-mm needle. Twenty-four hours after the biopsy each patient underwent routine ultrasound examination to exclude a possible bleeding. In eight cases an afterbleeding occurred (total hemorrhagic rate, 2.5%), four times without clinical consequences. The other four bleeding complications were more serious (1.2% of all taps), and all occurred after liver biopsies in patients with a history of liver complaints and abnormal clotting variables. There were no fatalities among our biopsies (mortality rate, 0%). CONCLUSION: The automatic Autovac biopsy system is suitable and relatively safe for obtaining sufficient histopathologic material from intra-abdominal organs. PMID- 10672842 TI - Vagal withdrawal during endoscopic retrograde cholangiopancreatography. AB - BACKGROUND: Patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) are at risk of developing cardiorespiratory complications, but the mechanism is still unknown. Treatment with metoprolol 2 h before the endoscopy has been shown to decrease the incidence of myocardial ischaemia during ERCP. The present study evaluated whether the endoscopic stress would decrease vagal tone and whether metoprolol given before the procedure could prevent this defence-like reaction. METHODS: Thirty-eight patients were randomized to receive either placebo or 100 mg metoprolol 2 h before ERCP. During ERCP the patients were monitored with a Holter tape recorder. Holter tapes from 31 patients (16 receiving metoprolol) were available to analyse the ratio of the standard deviations of the RR intervals (SDRR) to the mean RR intervals (measure of vagal tone) during ERCP. RESULTS: A decreased vagal tone during the ERCP was found, but we observed no difference between the metoprolol and the placebo group. For both groups the lowest vagal tone occurred at maximum heart rate during endoscopy. The SDRR/meanRR ratio returned towards base line for the subsequent 60 min after endoscopy. CONCLUSIONS: The existence of a defence-like reaction ('vagal withdrawal') during ERCP has been shown. Metoprolol given 2 h before the procedure did not affect the occurrence of this phenomenon. The interaction of other periendoscopic factors is still unclear and should be studied further. PMID- 10672844 TI - Actinomycosis of the colon: a rare form of presentation. AB - Actinomycosis is an uncommon entity, caused by an anaerobic bacterium, Actinomyces israelii, which is a component of the human oral and gastrointestinal flora. The cervicofacial region is the commonest site of disease, and the abdomen is the second commonest. In this situation the disease is almost always unifocal and restricted to the right colon, especially to the cecum. We report here the case of a patient with a very rare form of this entity, characterized by multiple foci of abdominal involvement with the most severe lesions localized in the transverse and sigmoid colon. The clinical presentation resembled a picture of colon perforation by cancer or diverticulitis, and the diagnosis was made by histopathologic examination of the lesions removed at surgery. No predisposing factor was found. The infection was successfully treated with a prolonged course of penicillin, after the surgical removal of the lesions. PMID- 10672845 TI - Food intolerance and gallstones: erratum and update of a meta-analysis. PMID- 10672846 TI - The role of Helicobacter pylori infection in gastroesophageal reflux disease. PMID- 10672847 TI - Tetrahydrobiopterin improves endothelial function in patients with coronary artery disease. AB - Tetrahydrobiopterin (BH4) is an essential cofactor for nitric oxide synthase (NOS) and a scavenger of oxygen-derived free radicals. Decreased availability of BH4 leads, under in vitro conditions, to reduced nitric oxide (NO) production and increased superoxide formation. We studied the effect of exogenous BH4 on endothelial function of angiographically normal vessel segments in patients with coronary artery disease. Nineteen patients with coronary artery disease underwent quantitative coronary angiography with simultaneous coronary flow velocity measurements (Cardiometrics FloWire). Data were obtained in angiographically normal segments of the left coronary artery at baseline, after intracoronary (i.c.) administration of acetylcholine (Ach; 10(-4) M), after infusion of BH4 (10(-2) M), and after co-infusion of ACh and BH4. At the end of the study, 300 microg nitroglycerin (NTG) i.c. was administered to obtain maximal vasodilation. At each step, flow velocity was determined before and after 18 microg adenosine i.c. to assess coronary flow velocity reserve. In 15 patients, ACh induced coronary vasoconstriction of -18 +/- 3% (endothelial dysfunction; p < 0.0001 vs. baseline), and in four patients, vasodilation of +39 +/- 20%. In the 15 patients with endothelial dysfunction, BH4 alone did not influence vessel area but prevented vasoconstriction to ACh (+2 +/- 3%, NS, vs. baseline). Correspondingly, calculated volume flow showed the highest value after co-infusion of ACh and BH4. Coronary flow velocity reserve was comparable during the various infusion steps. BH4 prevents ACh-induced vasoconstriction of angiographically normal vessels in patients with coronary artery disease. Thus substitution of this cofactor of NOS may represent a new approach for the treatment of endothelial dysfunction. PMID- 10672848 TI - Treatment of atherosclerosis in apolipoprotein E-deficient mice with 4-(3 Bromobenzoyl)-6,7-dimethoxyquinazoline (WHI-P164), a potent inhibitor of triglyceride synthesis. AB - We identified a novel organic compound, 4-(3'-bromobenzoyl)-6,7 dimethoxyquinazoline (compound WHI-P164), as a potent inhibitor of triglyceride (TG) synthesis. In an in vitro model of lipid synthesis, WHI-P164 (but not any one of the three structurally similar control dimethoxyquinazoline compounds) inhibited the accumulation of TG-rich intracellular lipid droplets in Caco-2 human intestinal cells in a concentration-dependent fashion. WHI-P164 caused no acute toxicity associated with morbidity or mortality in mice when administered at dose levels ranging from 0.5 to 80 mg/kg. In pharmacokinetic studies in mice, WHI-P164 was rapidly eliminated from plasma with a terminal elimination half-life of 26.1 +/- 1.3 min after intraperitoneal administration and 33.3 +/- 11.3 min after intravenous administration. Treatment with 40 mg/kg WHI-P164 (but not one of three structurally similar control dimethoxyquinazoline compounds) administered intraperitoneally once daily for 7 consecutive treatment days blocked the in vivo hepatic TG synthesis in both apoE-deficient and wild-type C57B1/6 mice. In apoE-deficient mice maintained on a high-fat/high-cholesterol Western diet, WHI-P164 substantially reduced the lipid accumulation in the liver after 7 days of treatment and the lipid accumulation in the aorta after 1 month of treatment. Our results in apoE-deficient mice show that lipid accumulation in hepatocytes and foam cells are related events, and inhibiting TG synthesis with WHI-P164 offers an effective means to treat atherosclerosis. PMID- 10672849 TI - Effects of adenosine receptor agonists on efferent renal nerve activity in anesthetized rats. AB - The aim of this study was to investigate the effects of A1 and A2 adenosine receptor activation on the sympathetic nervous system. The effects on efferent renal nerve activity of selective A1 (CCPA; 2-chloro-N-6-cyclopentyladenosine) and A2 (2HE-NECA; 2-hexynyl-5'-N-ethylcarboxamidoadenosine) adenosine-receptor agonists were studied in anesthetized rats either with intact baroreflexes (intact rats) or with bilateral sinoaortic denervation and vagotomy (denervated rats). After a control period of 5 min, A1 or A2 agonist or vehicle were intravenously infused for 8 min in separate groups of intact or denervated rats, in which arterial pressure and heart rate were continuously recorded. CCPA (5.0 microg/kg/min) and 2HE-NECA (0.7 microg/kg/min) were selected to obtain comparable blood pressure changes over the period of observation. Arterial pressure significantly and equally decreased during the A1 (-41 +/- 8%), and A2 ( 35 +/- 5%) agonist administration. Heart rate significantly decreased during A1 agonist infusion, but it did not change during A2 agonist administration. Bilateral sinoaortic denervation and vagotomy did not modify the hemodynamic responses to both drugs. The A1 and A2 administration caused a large and significant increase in efferent renal nerve activity (+66 +/- 22% and +76 +/- 15%, respectively), and this effect was entirely abolished in denervated rats. A linear relation with a significant negative slope between changes in arterial pressure and changes in neural discharge was observed for each treatment. The comparison of the regression slopes showed that the reflex increase of efferent sympathetic activity caused by the administration of both agonists was significantly smaller than the increment induced by equipotent hypotensive dose of sodium nitroprusside (10 microg/kg). These data show that the selective activation of A1 and A2 receptors elicits a reflex increase in efferent renal nerve activity. This neural activation is smaller as compared with the effect of equihypotensive doses of sodium nitroprusside, thus indicating a blunting effect of both adenosine agonists on baroreceptor sensitivity. PMID- 10672850 TI - Amlodipine enhances NO production induced by an ACE inhibitor through a kinin mediated mechanism in canine coronary microvessels. AB - Our previous study found that angiotensin-converting enzyme (ACE) inhibitors and amlodipine induce NO release from coronary microvessels through a kinin-dependent mechanism. The goal of this study was to determine whether amlodipine could potentiate NO formation during ACE inhibition. Coronary microvessels were isolated from 16 mongrel dogs. Nitrite, the hydration product of NO, from coronary microvessels was quantified by using the Griess reaction. Bradykinin and kallikrein all significantly increased nitrite release from coronary microvessels in a concentration-dependent manner. The ACE inhibitor, ramiprilat, potentiated these effects. Amlodipine also markedly potentiated nitrite production by ramiprilat. For instance, amlodipine (10(-10) M) enhanced nitrite release induced by ramiprilat (10(-7) M) from 122 +/- 9 to 168 +/- 14 pmol/mg (p < 0.05 vs. ramiprilat). Nitrite release potentiated by ramiprilat and amlodipine was entirely blocked by N(omega)-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase), HOE 140 (Icatibant, a specific B2-kinin receptor antagonist), and dichloroisocoumarin (DCIC, a serine protease inhibitor that blocks local kinin formation). These results clearly show that there is a synergistic effect on NO formation when amlodipine is combined with ACE inhibition. Our data suggest that kinin-mediated coronary NO production may contribute importantly to the beneficial therapeutic action of ACE inhibitors, especially in combination with amlodipine in the treatment of heart disease. PMID- 10672851 TI - The protein kinase inhibitor fasudil protects against ischemic myocardial injury induced by endothelin-1 in the rabbit. AB - Endothelin-1 (ET-1) induces severe pathologic conditions such as coronary spasm followed by vasospastic angina pectoris and acute myocardial infarction. The related pathophysiologic mechanisms have remained obscure. Endothelin-1 receptor (ET(A) and ET(B)) is reported to couple with several types of G protein-involved pathways that participate in phospholipase C activation and atrial myofibrils organization into sarcomeric units. Here we demonstrate that ET-1 induces histologic and pathologic dysfunction in the rabbit myocardium and that such pathologic events are prevented by the Rho-kinase inhibitor fasudil. Although the bolus injection of ET-1 (1.4 nmol/kg) via the auricular vein of the rabbit induced only transient T-wave elevation, irreversible, severe histologic changes were observed in papillary muscles of the ventricle, and multifocal myocardial necrosis with infiltration of neutrophils and macrophages in the left ventricle occurred. Oral administration of fasudil (10 mg/kg) significantly reduced the occurrence of myocardial injury determinants, whereas conventional Ca2+ channel blockers (nifedipine, diltiazem) and a K+ channel opener (nicorandil; 10 mg/kg, p.o. each) showed a lesser or no effect on such determinants. These results suggest that ET-1 induces severe myocardial dysfunction based not only on the occurrence of vasospastic ischemia but also on its direct effects on the myocardium. PMID- 10672852 TI - Methoxamine inhibits transient outward potassium current through alpha1A adrenoceptors in rat ventricular myocytes. AB - alpha1-Adrenoceptor agonists are known to reduce transient outward potassium current (I(to)) in the heart. The aim of this study was to analyze the effect of methoxamine (mtx) on I(to) and to elucidate which adrenoceptor subtype was involved in this effect. We used the whole-cell configuration of the patch-clamp technique to record I(to). Our experiments confirm that mtx induces a dose dependent decrease of I(to) that is characterized by an acceleration of time to peak (3.5 +/- 0.2 and 2.3 +/- 0.3 ms for control and mtx, respectively), and a decrease in both inactivation time constants (T(fast) was reduced from 20.8 +/ 2.6 to 14.9 +/- 1.1 ms, and tau(slow) was reduced from 138 +/- 32.1 to 114 +/- 28.7 ms; n = 7). All these effects were antagonized by prazosin and the alpha1A antagonist 5-methylurapidil but not by the irreversible alpha1B-antagonist chloroethylclonidine. These data indicate that stimulation of alpha1A adrenoceptor subtype is involved in the methoxamine-induced reduction of I(to) in rat ventricular myocytes. PMID- 10672853 TI - Nitroprusside reverses lengthened time of contraction in stunned canine cardiac myocytes. AB - We tested the hypothesis that stunning reduces the function of isolated canine ventricular myocytes and that nitroprusside (NP) reverses this effect. After stunning (15 min occlusion, 45 min reperfusion), isolated myocytes were prepared from control (circumflex artery) and stunned (left anterior descending) regions of the hearts of seven dogs. The myocytes were examined at baseline and with NP (10(-6,-5,-4) M) for oxygen consumption (MVO2, nl O2/min/10(5) cells), cyclic guanosine monophosphate (cyclic GMP; fmol/10(5) cells), and cell contraction. Basal MVO2 was not significantly different between control and stunned myocytes (888 +/- 108 vs. 716 +/- 94). NP caused a dose dependent decrease in MVo2 (control, 262 +/-51; stunned, 287 +/- 59, NP 10(-4) M). Basal cyclic GMP levels were comparable between control and stunned myocytes (117 +/-28 vs. 124 +/- 18). NP produced a similar dose-dependent increase in cyclic GMP in control and stunned myocytes. Baseline cell shortening (%) was similar in control vs. stunned myocytes (12.1 +/- 1.2 vs. 11.0 +/- 0.9). NP reduced shortening (6.9 +/- 0.3 vs. 7.3 +/- 0.5, NP 10(-4) M). There was no baseline difference in maximal rate of shortening (microm/s) between control and stunned myocytes (164 +/- 14, 157 +/- 20). With NP, a decrease in the maximal rate of shortening was seen in both groups (128 +/- 12, 139 +/- 21, NP 10(-4) M). The time of contraction (s) was significantly longer in stunned (0.20 +/- 0.03) versus control (0.13 +/- 0.01). NP significantly lengthened the time of contraction in controls in a dose dependent manner (0.33 +/-0.05, NP 10(-4) M). In stunned myocytes, however, low dose NP (10(-6) M) caused a decrease in the time of contraction (0.15 +/-0.03). High-dose NP (10(-4) M) did not significantly lengthen time of contraction in stunned cells (0.23 +/- 0.02). The time of relaxation followed a similar pattern. We conclude that part of the effect of NP in low doses in stunned myocardium is to reduce the lengthened time of contraction and relaxation characteristic of stunning. PMID- 10672854 TI - Genistein potentiates the relaxation induced by beta1- and beta2-adrenoceptor activation in rat aortic rings. AB - In rat aortic rings, genistein, an inhibitor of tyrosine kinase, but not daidzein, an inactive analogue of genistein, potentiated the relaxation induced by isoproterenol. Atenolol, a beta1-adrenoceptor antagonist, or ICI-118,551, a beta2-adrenoceptor antagonist, inhibited the relaxation induced by isoproterenol. The potentiating effect of genistein on the relaxation induced by isoproterenol in the presence of ICI-118,551 was apparently greater than that in the presence of atenolol. In the presence of ICI-118,551, theophylline, an inhibitor of cyclic adenosine monophosphate (cAMP)-dependent phosphodiesterase (cAMP-PDE), markedly inhibited the potentiating effect of genistein on the isoproterenol-induced relaxation, whereas in the presence of atenolol, theophylline only partly inhibited the potentiating effect of genistein. The relaxation induced by forskolin, an activator of adenylyl cyclase, was potentiated by genistein or theophylline. In the presence of theophylline, the relaxation induced by forskolin was not further affected by genistein. Genistein also inhibited the activities of cAMP-PDE. In the presence of atenolol, but not ICI-118,551, iberiotoxin, an inhibitor of Ca-activated K channels, inhibited the relaxation induced by isoproterenol and the potentiating effect of genistein. In the presence of atenolol, quinacrine, an inhibitor of phospholipase A2, and metyrapone, an inhibitor of P-450 enzymes, but not alpha-naphthoflavone, an inhibitor of P-450 enzymes, indomethacin, a cyclooxygenase inhibitor, or AA861, a 5-lipoxygenase inhibitor, inhibited the potentiating effect of genistein. These results suggest that the potentiation of the beta1-adrenoceptor-induced relaxation by activation of genistein may mostly be due to inhibition of cAMP-PDE activities. In addition, the potentiation of the relaxation induced by activation of beta2-adrenoceptors by genistein may be related to the inhibition of arachidonic acid metabolism and cAMP-PDE activities. PMID- 10672856 TI - Beneficial effects of the T- and L-type calcium channel antagonist, mibefradil, against exercise-induced myocardial stunning in dogs. AB - Abnormalities in calcium homeostasis such as calcium overload have been shown to participate in the pathogenesis of myocardial stunning. The goal of this study was to investigate the effects of mibefradil, a mixed T- and L-type calcium channels antagonist on exercise-induced ischemia (i.e., high-flow ischemia). Nine dogs were permanently instrumented to measure left ventricular wall thickening (Wth) and coronary blood flow (Doppler). Infusion of saline or mibefradil (30 and 40 microg/kg/min, i.v., for 20 min) was started 10 min before exercise (10 min, 14 km/h; slope, 13%) and stopped at its end. Circumflex coronary artery stenosis (pneumatic occluders) was set up 5 min before exercise to suppress exercise induced increase in mean coronary blood flow without simultaneously affecting Wth at rest. Mibefradil (30 microg/kg/min) was also administered at the beginning of the recovery period in a subset of four dogs. During exercise with saline, Wth was dramatically reduced (-77 +/- 7%; p < 0.05) and recovered only after 24 h. Mibefradil at both doses significantly limited tachycardia during exercise (211 +/- 7 and 210 +/- 5 beats/min vs. 240 +/- 8 beats/min for mibefradil, 30 microg/kg/min, mibefradil, 40 microg/kg/min, and saline, respectively) but exerted no negative inotropic effects. Mibefradil at both doses significantly reduced the intensity of myocardial stunning and the time to recovery in Wth (3 h). Administration of mibefradil at the beginning of the recovery period did not protect against myocardial stunning. Administration of a mixed T- and L-type calcium channel antagonists before ischemia confers cardioprotection against exercise-induced myocardial stunning. This may potentially be related to the limitation of exercise-induced tachycardia and/or the prevention of calcium overload. PMID- 10672855 TI - Butanolic fraction from Cuphea carthagenensis Jacq McBride relaxes rat thoracic aorta through endothelium-dependent and endothelium-independent mechanisms. AB - This study evaluated the vasorelaxant properties of the crude hydroalcoholic extract (CE) of Cuphea carthagenensis, as well as its butanolic (BF) and ethyl acetate (EA) fractions, in rings of rat thoracic aorta. In endothelium-intact rings contracted with phenylephrine (30-100 nM), cumulative additions of increasing concentrations of CE, BF, and EA of C. carthagenensis (0.1 microg/ml-3 mg/ml) caused graded relaxations, with BF displaying the lowest median inhibitory concentration (IC5; mean, 6.8 microg/ml; 95% confidence limits, 3.3-14.2). BF induced relaxations of endothelium-intact rings were virtually abolished by prior incubation with the NO-synthase inhibitor N(omega)-nitro-L-arginine (L-NOARG; 10 or 30 microM), and were markedly reduced after guanylate cyclase inhibition with either methylene blue (10 microM) or ODQ (1 microM; 1H[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one). The inhibition of BF-induced relaxation by L-NOARG was prevented to a large extent by simultaneous incubation with L-arginine (1 mM). In endothelium-denuded rings contracted with phenylephrine, CE and BF caused graded relaxations only at doses >100 microg/ml, whereas the NO-donors SNAP (S-nitroso-N acetyl-penicillamine) and SIN-1 (3-morpholino-sydnonimine) induced full relaxation at 1 microM. BF (100 microg/ml), which caused little relaxation per se of endothelium-denuded rings, potentiated the relaxant effects of SNAP and even more so of SIN-1 (which, unlike SNAP, also releases superoxide anion O2- in addition to NO), in a manner qualitatively similar to that seen with SOD (superoxide dismutase) against SIN-1. These data indicate that the BF of C. carthagenensis induces relaxation of the rat thoracic aorta by two mechanisms: (a) an endothelium-dependent component of action, which clearly depends on the NO/cyclic guanosine monophosphate (cGMP) pathway and can be attributed, at least in part, to free radical-scavenging properties; and (b) an endothelium independent component of action, which becomes evident at higher doses (> or = 100 microg/ml) and remains to be further characterized. These results suggest that this native South American plant might be beneficial in cardiovascular disease. PMID- 10672857 TI - Intracoronary enalaprilat improves metabolic coronary vasodilation in patients with idiopathic dilated cardiomyopathy. AB - Coronary flow reserve is reduced in patients with idiopathic dilated cardiomyopathy (DCM). We examined acute effects of intracoronary enalaprilat on metabolic coronary vasodilation during pacing tachycardia in patients. Coronary blood flow (Doppler guidewire) and diameter (quantitative angiography) were measured in seven patients with DCM and seven control subjects. In the DCM group, tachypacing increased coronary blood flow by 37 +/- 22% from the baseline before enalaprilat and by 65 +/- 22% (p < 0.01 vs. before treatment) after enalaprilat (0.5 microg/kg/min for 5 min, i.c.) at comparable double product. Pacing-induced dilation of the epicardial coronary artery also was greater after enalaprilat (p < 0.05). Effects of enalaprilat on coronary blood flow and diameter during pacing tachycardia were abolished by pretreatment with intracoronary administration of the nitric oxide (NO) synthesis inhibitor, N(G)-monomethyl-L-arginine. These beneficial effects of enalaprilat on large and small coronary vasodilation were not observed in control patients. Thus, intracoronary enalaprilat acutely augmented dilator responses of the large and small coronary arteries to pacing tachycardia in patients with DCM, and NO appeared to play an important role in mediating the effects of enalaprilat. These favorable effects of enalaprilat on the coronary circulation may be of clinical significance in patients with heart failure due to nonischemic DCM. Further long-term studies of the effects of angiotensin-converting enzyme inhibition on coronary vasodilation will be needed in this population. PMID- 10672858 TI - Lipophilic HMG-CoA reductase inhibitors increase myocardial stunning in dogs. AB - Pretreatment of dogs with simvastatin, a lipophilic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, increases myocardial contractile dysfunction during reperfusion after ischemia (stunning), with reduction of tissue adenosine triphosphate (ATP). This was thought to be a consequence of prevention of ubiquinone biosynthesis by the lipophilic inhibitor in the myocardial cell. We examined whether other lipophilic HMG-CoA reductase inhibitors also influence myocardial stunning in dogs. Vehicle, atorvastatin (2 mg/ kg/day), fluvastatin (4 mg/kg/day), or cerivastatin (40 microg/kg/ day) was orally administered for 3 weeks. Hydrophilic pravastatin (4 mg/kg/day) also was given. After 3 weeks, pentobarbital-anesthetized dogs were subjected to 15-min left anterior descending coronary artery occlusion followed by 2-h reperfusion. Myocardial segment function was determined by sonomicrometry. Tissue levels of ATP were determined in 2-h reperfused hearts. All inhibitors significantly decreased serum cholesterol level. The three lipophilic inhibitors resulted in a worsening of segment function in the reperfused myocardium, as compared with the vehicle group. The levels of ATP in the atorvastatin, fluvastatin, and cerivastatin groups were significantly lower than that in the vehicle group. These results confirm that lipophilic HMG-CoA reductase inhibitors enhance myocardial stunning in association with ATP reduction after ischemia and reperfusion. PMID- 10672859 TI - Myocardial protection with red wine extract. AB - Cardioprotective action of red wine was studied by preperfusing isolated rat hearts with ethanol-free red wine extract for 15 min before subjecting them to 30 min of global ischemia followed by 2 h of reperfusion. Four other group of rats were studied under identical conditions, of which one served as control; one was treated with 10 microM trans-resveratrol (RVT), one of the major antioxidants found in red wines; another, with 0.07% ethanol; and another, with 0.07% ethanol plus 10 microM RVT. The results of our study demonstrated that both red wine extract and RVT were equally cardioprotective, as evidenced by their abilities to improve postischemic ventricular functions including developed pressure and aortic flow. Developed pressure values at 60 min after reperfusion were 81.8 +/- 1.2 and 68.8 +/- 4.1 mm Hg for the red wine extract and RVT groups, respectively, versus 49.7 +/- 2.7 mm Hg for the control group. These compounds also reduced myocardial infarct size compared with the control hearts (20.1 +/- 0.5% and 10.5 +/- 0.3% for red wine extract and RVT groups, respectively, vs. 29.9 +/- 3.1% for the control group). The ethanol-treated group displayed slightly better functional recovery, which deteriorated sharply toward the end of the reperfusion period, and the extent of infarction was comparable to that of the control group (31.5 +/- 0.9%). In the ethanol plus RVT group, postischemic contractile function was significantly better than control, and infarct size also was reduced to 20.9 +/- 0.7%. The amount of malonaldehyde formation in the postischemic myocardium was reduced by red wine extract and RVT, indicating a reduction of oxidative stress developed in the ischemic reperfused myocardium. In vitro studies revealed that red wine extract is a potent antioxidant as evidenced by its ability to scavenge peroxyl radical in vitro. Taken together, the results of our study indicate that red wines are cardioprotective by their ability to function as an in vivo antioxidant. PMID- 10672860 TI - A peripheral site of action for the attenuation of baroflex-mediated bradycardia by intravenous mu-opioid agonists. AB - We previously reported that i.v. DAMGO (Tyr-D-Ala-Gly-NMePhe-Gly-ol), a selective mu-opioid agonist, causes an increase in blood pressure with no change in heart rate in unanesthetized sheep and subsequently demonstrated that DAMGO attenuates baroreflex-mediated bradycardia. To determine the site and mechanism by which mu agonists inhibit baroreflex sensitivity, we have carried out further investigations by using DAMGO and another mu-agonist, DALDA (Tyr-D-Arg-Phe-Lys NH2). The bradycardic response to norepinephrine (NE) was significantly blunted after i.v. DAMGO or DALDA in both nonpregnant and pregnant sheep. In contrast, the tachycardic response to sodium nitroprusside (SNP) remained unchanged in the presence of DAMGO or DALDA. In view of the highly restricted distribution of DALDA across the blood-brain barrier (BBB), we hypothesized that the blunting of reflex-mediated bradycardia by mu-opioid agonists can occur peripherally. Pretreatment with the quaternary opioid antagonist, naloxone methiodide (NM), completely blocked the attenuation of baroreflex sensitivity by DAMGO and DALDA in both nonpregnant and pregnant animals. These data suggest that in addition to central mechanisms, mu-opioid agonists can inhibit baroreflex sensitivity at a peripheral site, most likely by inhibiting vagal influence on heart-rate control rather than by acting directly at baroreceptors. PMID- 10672861 TI - Insulin-like growth factor-I improves recovery of cardiac performance during reperfusion in isolated rat heart by a wortmannin-sensitive mechanism. AB - Insulin-like growth factor-I (IGF-I) has been shown to produce a short-term positive inotropic effect (PIE) in the myocardium under nonischemic conditions. IGF-I also conferred cytoprotection against ischemia and reperfusion injury in various organs. IGF-I may, therefore, facilitate the recovery of postischemic cardiac function. Isolated and crystalloid-perfused rat heart was subjected to 25 min of normothermic ischemia followed by 30 min of reperfusion. IGF-I produced PIE in a dose-dependent manner at concentrations ranging between 1 and 100 nM under nonischemic conditions. Although 1 nM isoproterenol produced much greater PIE and myocardial energy conversion efficiency (MECE) than did 65 nM IGF-I in this condition, the same concentration of IGF-I administered during reperfusion conferred better recovery of left ventricular function and MECE compared with isoproterenol. The improved cardiac performance by IGF-I was associated with lower release of creatine kinase (CK). Wortmannin (100 nM), a specific inhibitor of phosphatidylinositol kinase (PI-3 kinase), abrogated IGF-I-induced improvement of contractile function and inhibition of CK release in the postischemic heart. We conclude that IGF-I administered during reperfusion accelerates recovery of cardiac performance and mitigates myocardial injury through a wortmannin sensitive mechanism. PMID- 10672862 TI - The role of cromakalim and a nitric oxide synthase blocker in cardiac arrhythmia in the intact baboon model. AB - The arrhythmogenic effect of adenosine triphosphate (ATP)-sensitive potassium channel openers is controversial and may be dependent on the type of animal model used. Information on the effect of these drugs in the normal primate model is limited. The purpose of this study was first to determine the arrhythmogenic properties of cromakalim in the baboon and second to determine whether N-omega nitro-L-arginine methyl ester (L-NAME) has any effect on the induced arrhythmia. Adult (2-4 years old) baboons (Papio ursinus) were anesthetized with a continuous i.v. infusion of ketamine (100 mg/ ml), diazepam (5 mg/ml), and saline (ratio 2:2:50) at a rate of 40-60 ml/h. Sympathetic responses were inhibited by administration of propranolol (1 mg/kg) before the start of the experiments. Cromakalim (30 microg/kg) was administered before and after L-NAME (7.5 mg/kg), and the parameters were monitored for 15 min after each intervention. A Millar double-tipped microcatheter was used to record left ventricular and aortic pressures. Lead II of the ECG was monitored. During a 15-min period after administration of cromakalim, 22.3 +/- 6.0 abnormal ventricular complexes were recorded. L-NAME administration significantly reduced these abnormal complexes to 4.5 +/- 2 (paired t test, p < or = 0.05). We therefore conclude that cromakalim has arrhythmogenic properties in the baboon and that these can be attenuated by L NAME. PMID- 10672863 TI - Effects of the K+ channel opener KRN4884 on the cardiovascular metabolic syndrome model in rats. AB - We examined the effects of the potassium channel opener KRN4884 (5-amino-N-[2-(2 chlorophenyl)ethyl]-N'-cyano-3-pyridinecarboxamidine ) on cardiovascular metabolic syndrome (i.e., syndrome X), in rats. High-fructose diet rats developed hypertension, hypertriglyceridemia, increased total cholesterol/HDL (high-density lipoprotein)-cholesterol ratio, and hyperinsulinemia, KRN4884 (0.3-3.0 mg/kg, twice a day for 14 days, p.o.) alleviated the risk factors in fructose-fed rats. Furthermore, fructose-fed rats exhibited impairment of glucose tolerance and excess insulin secretion when loaded with glucose orally. Treatment with KRN4884 (1.0 mg/kg, twice a day for 14 days, p.o.) improved the glucose intolerance and inhibited hypersecretion of insulin in the glucose-loaded, fructose-fed rats. In contrast, KRN4884 (0.3-1.0 mg/kg, twice a day for 10 days, p.o.) did not affect serum triglyceride, cholesterol, glucose, or insulin concentrations in normal rats. LPL (lipoprotein lipase) activities in skeletal muscle and adipose tissue, and HTGL (hepatic triglyceride lipase) activity in liver were measured after administration of KRN4884 or vehicle twice a day for 14 days in fructose-fed rats. KRN4884 caused a significant increase in LPL activity in muscle and tended to increase LPL activity in adipose tissue in fructose-fed rats. HTGL was decreased in fructose-fed rats as compared with normal controls and was unaffected by KRN4884. These findings suggested that KRN4884 enhances insulin sensitivity and LPL activity, which are related to glucose and lipid metabolism and may be useful for the treatment of syndrome X. PMID- 10672864 TI - Sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, inhibits serotonin-induced coronary artery spasm in a porcine model. AB - Serotonin is one of the most important vasoactive substances and has been implicated in the pathogenesis of coronary artery spasm and of acute coronary syndrome. We have recently demonstrated that local and long-term treatment with interleukin-1beta(IL-1beta) causes coronary arteriosclerotic changes and hyperconstrictive responses to serotonin in pigs in vivo. However, it remains to be examined which serotonergic (5-HT) receptor subtype mediates coronary spasm and whether alterations in serotonergic receptors are involved in the abnormality. In this study, we examined the inhibitory effect of sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, on the serotonin-induced coronary spasm as well as the possible alterations of serotonergic receptors in our porcine model. A segment of the porcine coronary artery was carefully dissected and aseptically wrapped with cotton mesh absorbing IL-1beta-bound microbeads from the adventitia. Two weeks after the procedure, angiographic study was performed, followed by binding assay for 5-HT1B and 5-HT2A serotonergic receptors and reverse transcription-polymerase chain reaction (RT-PCR) analysis for mRNA of those receptors. Angiographic study showed coronary vasospastic responses to serotonin at the IL-1beta-treated site. Sarpogrelate dose dependently inhibited the serotonin-induced coronary spasm, but it did not affect the prostaglandin F2alpha-induced vasoconstriction. Radiolabeled receptor-binding assay showed that receptor affinity or receptor number of the 5-HT1B, or 5-HT2A receptors did not differ significantly between the spastic and the control sites. Furthermore, RT-PCR analysis showed that the expression of neither 5-HT2A nor 5 HT1B receptor mRNA was significantly altered at the spastic site. These results indicate that serotonin-induced coronary spasm is mediated primarily by 5-HT2A receptor in our porcine model, although the 5-HT2A receptor was not up-regulated, suggesting that alteration in the signal-transduction pathway for vascular smooth muscle contraction beyond the 5-HT2A receptor plays a primary role in the pathogenesis of coronary spasm in our porcine model. PMID- 10672865 TI - RAR gamma agonists inhibit proliferation of vascular smooth muscle cells. AB - The multifactorial and unpredictable nature of human restenosis will probably necessitate interventional strategies that target multiple processes involved in neointimal proliferation. Retinoids represent a growing class of pleiotropic biologic response modifiers with demonstrable efficacy in managing several pathologic conditions pertaining to neointimal proliferation. However, retinoid treatment is associated with a high incidence of adverse effects. The action of all-trans-retinoic acid is mediated by two families of nuclear receptors, RARs and RXRs, each containing three isoforms alpha, beta, and gamma. Because synthetic retinoids that are receptor and function specific have been shown to differ from each other by several orders of magnitude in their potencies and are associated with limited adverse effects, we examined the effect of synthetic retinoids on serum- and serotonin-induced vascular smooth muscle cell (VSMC) proliferation. Naturally occurring retinoids were used as controls. All-trans retinoic acid at nanomolar concentrations inhibited smooth muscle cell proliferation. In this study, we report that RAR gamma subgroup-specific agonists are the most potent inhibitors of serum and serotonin VSMC proliferation, as compared with other RAR pan-agonists and naturally occurring retinoids tested. Our results indicate that RAR gamma subgroup-specific agonists should be assessed further in in vivo models of neointimal proliferation. PMID- 10672866 TI - Efficacy of the novel calcium antagonist R(+)-semotiadil in limiting the ventricular rate during atrial flutter in isolated guinea pig hearts. AB - Ca2+ antagonists slow the ventricular rate by blocking conduction in the anterograde direction through the atrioventricular (AV) node. The aim of this study was to investigate the efficacy of the novel Ca2+ antagonist semotiadil compared with verapamil and diltiazem on the filtering capacity of the AV node during simulated atrial flutter in isolated guinea-pig hearts perfused by the method of Langendorff. During sinus rhythm, semotiadil as well as verapamil and diltiazem induced comparable depressant effects on AV-nodal conduction time and, during tachycardia, a comparable enhancement of this effect. The time constant (tau-on) for the drug-specific rate-dependent effect on AV-nodal conduction slowing was longest in the presence of verapamil compared with the long tau-on of semotiadil and the short tau-on of diltiazem. Verapamil and semotiadil exhibited a significantly greater effect than diltiazem on the mean ventricular cycle length (VCLmeun), on the maximal ventricular cycle length (VCLmax) and on the standard deviation of the VCL (SD(VCL)) during atrial flutter. Therefore the kinetics of the rate adaptation of AV-nodal conduction time in the presence of Ca2+ antagonists predicts the filtering capacity of the AV node during atrial flutter. Semotiadil has a verapamil type of action on ventricular cycle length during atrial flutter, whereas the disadvantageous prolongation of maximal VCL as well as the dispersion of VCL with semotiadil was only about half those found with verapamil. PMID- 10672867 TI - Role of NPY Y1 receptors in cardiovascular control in the conscious rabbit. AB - Prejunctional neuropeptide Y (NPY) Y1 receptors on cardiac sympathetic neurons mediate transient inhibition of chronotropic responses in rabbit isolated right atria. The function of these receptors remains speculative. We investigated a possible functional role for these receptors in modulation of the baroreceptor heart rate (HR) reflex in the conscious rabbit. Mean arterial pressure (MAP) responses to a range of doses of the Y1 receptor agonist [Leu31,Pro34]NPY (1-8 microg/kg, i.v.) were constructed in ganglion-blocked rabbits. After administration of the selective Y1 receptor antagonist GR231118(150 microg/kg, i.v.), two-point [Leu31,Pro34]NPY dose-pressor responses were assessed. Linear regression analysis of the relation between the shift in the [Leu31,Pro34]NPY dose-pressor response lines against time was used as an estimate of the functional half-life of GR231118. GR231118 shifted the two-point [Leu31,Pro34]NPY dose-pressor response relation by 10- to 30-fold. A single estimate of the functional half-life of a bolus dose of GR231118 was 25 +/- 2 min. This determination allowed a steady-state Y1-receptor blockade to be established by a bolus and infusion. In a separate group of rabbits, the baroreceptor-HR reflex was assessed before and 30 min after administration of GR231118 (150 microg/kg bolus, then 150 microg/ kg/h, i.v.). GR231118 caused an initial transient pressor response and bradycardia, followed by a depressor response and a more sustained tachycardia. Infusion of GR231118 had no effect on the baroreceptor-HR reflex. Prejunctional Y1 receptors appear not to mediate a tonic inhibition of cardiac sympathetic neurotransmission in the conscious rabbit during physiological manipulations in MAP. However, activation of postjunctional Y1 receptors by neuronal or circulating NPY may be important in maintenance of vascular tone in the conscious rabbit. PMID- 10672868 TI - Antagonism of coronary artery relaxation by adenosine A2A-receptor antagonist ZM241385. AB - We have tested the existence of functional A2A adenosine receptor in porcine coronary artery using, for the first time, the new A2A antagonist ZM241385. Nonselective agonist NECA and A2A-selective agonist CGS21680 produced concentration-dependent relaxation of prostaglandin F2alpha (PGF2alpha) precontracted endothelium intact (E+) and denuded (E-) rings. Relaxation was significantly greater in E+ rings than in E-rings. A2A adenosine receptor selective antagonist, ZM241385 (10(-6) M), significantly attenuated the relaxation responses. The antagonism of ZM241385 was compared with that of SCH58261 (10(-6)M), another A2A adenosine receptor-selective antagonist, which also significantly attenuated the relaxation response to both agonists. However, ZM241385 produced a significantly greater shift of the relaxation-response curves to the right compared with SCH58261 both in E+ and E- rings. The data show for the first time that ZM241385 is a potent A2A-receptor antagonist in porcine coronary artery and a useful tool to study A2A-receptor function. PMID- 10672869 TI - Relaxation to flavones and flavonols in rat isolated thoracic aorta: mechanism of action and structure-activity relationships. AB - The mechanism of the relaxant action and the structure-activity relation of flavonols (fisetin, quercetin, and 3,3',4'-trihydroxyflavone) and flavones (apigenin, chrysin, and luteolin) were examined in rat isolated thoracic aorta. The control responses to flavonols and flavones were compared with responses observed after the removal of the endothelium or in the presence of the L-type Ca2+ channel blocker, nifedipine (10(-7) M). The effects of flavonoids on contraction caused by the influx of extracellular Ca2+ and agonist-induced release of intracellular Ca2+ also were investigated. The flavones exhibited endothelium-independent vasorelaxation, whereas the removal of the endothelium significantly decreased the sensitivity of the relaxant responses to the flavonols without affecting the maximal relaxation. In the presence of nifedipine, the responses to apigenin, luteolin, and quercetin were significantly inhibited, but relaxation to chrysin, fisetin, and 3,3',4'-trihydroxyflavone was unaffected. All flavonols and flavones caused concentration-dependent inhibition of the contractile responses to exogenous application of Ca2+ and the release of intracellular Ca2+ stimulated by phenylephrine. Of the six flavonoids examined, 3,3',4'-trihydroxyflavone was the most potent when causing vasorelaxation or inhibition of contraction caused by the influx or release of Ca2+. In conclusion, these studies provide evidence that the hydroxyl substitution in the carbon 3 position that characterizes the flavonols is important in stimulating endothelium dependent vasorelaxation, and the absence of hydroxyl substitution on the A phenolic ring enhances the relaxant action. PMID- 10672870 TI - Analysis of the electrophysiologic effects of short-term oxybutynin on guinea pig and rabbit ventricular cells. AB - The objective of this study was to investigate the cardioactive properties of oxybutynin, a drug that is widely prescribed for management of voiding dysfunction. Membrane currents were recorded from whole-cell-configured guinea pig ventricular myocytes, and action potentials were recorded from guinea pig and rabbit papillary muscles. L-type Ca2+ current (I(Ca),L), inward-rectifier K+ current (I(K1)), and delayed-rectifier K+ current (I(K)) were unaffected by < or = 1 microM oxybutynin, and inhibited by higher concentrations. The concentrations that reduced the currents to one-half of predrug control amplitude (K0.5) were as follows: 1(Ca),L, 16.1 microM, I(K1), 18.2 microM, rapidly activating I(K)(I(Kr)), 11.4 microM, and slowly activating I(K)(I(Ks)), 28.7 microM. Action potential durations at 20 and 90% repolarization (APD20, APD90) were unaffected by oxybutynin < or =3 microM in guinea pig papillary muscles driven at 1 Hz; higher concentrations selectively shortened the APD20 by as much as 25% (100 microM), and caused moderate reductions in maximal upstroke velocity. Changes in the action potentials of rabbit papillary muscles were even smaller than in the guinea pig muscles. Because the peak therapeutic plasma concentration of oxybutynin is in the 0.01-0.1 microM range, the results suggest that the drug is highly unlikely to have adverse effects on cardiac electrical activity. PMID- 10672871 TI - Perillyl alcohol, an inhibitor of geranylgeranyl transferase, induces apoptosis of immortalized human vascular smooth muscle cells in vitro. AB - The isoprenoids geranylgeraniol and farnesol are required for lipid modification of several important cellular proteins. In this study the effect of perillyl alcohol (PA), an inhibitor of geranylgeranyl transferase, was examined on proliferation and apoptosis of immortalized human vascular smooth muscle cells (HVSMCs) derived from saphenous vein. PA induced a time- and concentration dependent inhibition of cell proliferation over 3 days and 48 h exposure to PA inhibited [methyl-3H-thymidine incorporation induced by 10% fetal calf serum in a concentration-dependent manner. Flow cytometry analysis indicated that PA reduced the proportion of cells in S phase and increased apoptosis. PA-induced apoptosis was confirmed by terminal deoxynucleotidyl transferase (TdT) reagent-based immunocytochemistry. In contrast, the selective inhibitors of farnesyl transferase B581 and BZA-5B were without effect. In view of the proposed role of proliferation and apoptosis in myointimal hyperplasia, inhibitors of geranylgeranyl transferases may have therapeutic potential in cardiovascular disease. PMID- 10672872 TI - Design of computer-generated hologram with ring focus for nonmechanical corneal trephination with Er:YAG laser in penetrating keratoplasty. AB - PURPOSE: To calculate a beam-shaping optical element for homogeneous intensity distribution within a focal ring to be used in nonmechanical trephination with the Er:YAG laser in penetrating keratoplasty instead of a spot guiding device. METHODS: The phase distribution behind a holographic optical element (HOE) k psi(u) can be described by the addition of the hologram phase phiH(u) to the beam phase phiE(u): k psi(u) = phiH(u) + phiE(u), k = 2pi/lambda, where u denotes the coordinates inside the hologram aperture, k an integer, and lambda the laser wavelength. To avoid discontinuous wavefronts leading to speckle noise, a smooth phase function is necessary. After transforming the hologram aperture coordinates into the focal plane x in a focal distance f, psi can be retrieved from the slope equation: inverted delta psi(u) = x(u) - u/f. RESULTS: Creating a ring focus can be reduced to an essentially one-dimensional problem by separation of variables due to the symmetry condition. We calculated a computer-generated eight-level phase-only HOE with 4096 x 4096 pixels from a Gaussian-distributed 2.94 Er:YAG laser spot with a beam diameter of 10 mm and a focal distance of 100 mm. Thereby, a ring focus with an inner/outer radius of 7/8 mm can be created. To avoid Poisson's spo, the symmetry of the problem was broken by circular modulation of the phase leading to a spiral-like structure. The calculated efficiency of the HOE relating the energy within the ring to the total energy was 91%. CONCLUSION: With an HOE it is possible to redistribute the energy along the desired focal ring. The HOE design can be adapted to the intensity distribution of the impinging laser beam with its characteristic aperture shape. A circular homogeneous corneal trephination depth is possible, because the energy fluctuation from pulse to pulse does not locally affect the ablation process. A ring focus for the Er:YAG laser has the potential to render superfluous a manual beam control via micromanipulator and to allow a more rapid and more regular corneal trephination along aperture masks. PMID- 10672873 TI - Expression of stress-response protein 60 in iritis associated with experimental autoimmune encephalomyelitis. AB - PURPOSE: To study the expression of stress-response proteins in the inflamed iris of rats with experimental autoimmune encephalomyelitis (EAE). METHODS: EAE was induced in Lewis rats by immunization with homogenized spinal cord of the guinea pig emulsified in complete Freund's adjuvant (CFA) (group EAE). Control rats included those immunized with only CFA (group CFA) and those that were untreated (group Normal). Immunohistochemical study for the localization of stress-response protein (srp) 27, srp 60, srp 72, ubiquitin, and alphaB-crystallin was performed. RESULTS: All rats in group EAE developed iritis, whereas none of the rats in group CFA and group Normal developed iritis. No expression of ubiquitin, alphaB crystallin, srp 27, srp 60, or srp 72 was seen in the epithelium of the iris in group CFA rats. In the eyes of rats in group EAE, srp 60 was expressed in the epithelium of the iris in 20 of 22 (90.9%), ubiquitin in 4 of 22 (18.2%), and alphaB-crystallin in 3 of 22 (13.6%). In the group Normal rats, only ubiquitin was expressed in the epithelium of the iris in 1 of 6 (16.7%) eyes examined. CONCLUSIONS: These results suggest that srp 60 may be a potential uveitogenic antigen in the iris in EAE. PMID- 10672874 TI - Prior topical anesthesia reduces time to full cycloplegia in Chinese. AB - PURPOSE: To investigate the effect of prior anesthesia on the time to full cycloplegia in young Chinese subjects. METHODS: The amplitude of accommodation was monitored over a 50-minute interval after the application of 1% cyclopentolate hydrochloride with a pretreatment of 0.4% benoxinate (oxybuprocaine) or 0.9% saline solution (control). Using a nonlinear mathematical model, the rate of accommodative loss (k) and the time required for 95% of total cycloplegia (T95%) were determined. RESULTS: Statistical analysis revealed a significantly faster rate of accommodative loss (P < .0001) after prior anesthesia (0.129 +/- 0.05) compared with the controls (0.103 +/- 0.04). T95% was noted at 26.43 +/- 10.22 minutes after prior anesthesia, which was significantly shorter (P < .0001) than that after the saline treatment (35.28 +/- 16.51 minutes). CONCLUSIONS: Prior application of topical anesthetic can shorten the time to full cycloplegia for people, such as the Chinese, with dark irides. PMID- 10672875 TI - Sorsby's fundus dystrophy in two Japanese families with unusual clinical features. AB - PURPOSE: To describe two Japanese families with Sorsby's fundus dystrophy (SFD) with unusual clinical features. METHODS: Two families from Kagoshima Prefecture with senile-onset macular dystrophy were examined. Three affected individuals through three successive generations of one family and three affected siblings in another family were examined and followed. RESULTS: The initial symptom of these patients was a rapid or slow central visual loss that occurred at an average age of 67.4 years. The major ophthalmoscopic changes consisted of soft drusen and hemorrhagic or atrophic lesions in the macula, which were progressive and ultimately led to disciform scarring. They had no difficulty with night vision. All the patients had normal peripheral retina with intact peripheral fields. They maintained good ambulatory vision and could walk unguided until late in life. These patients had a novel mutation in the tissue inhibitor of the metalloproteinases-3 (TIMP3) gene. CONCLUSIONS: This is the first report of SFD from the East. Its clinical features differ from those of SFD patients of the West, appearing closer to features of age-related macular degeneration. These two unrelated Japanese families with an identical mutation in the TIMP3 gene might be descendants of a common ancestor who carried the mutant gene. PMID- 10672876 TI - Effects of stimulus blocking, light scattering, and distortion on multifocal electroretinogram. AB - PURPOSE: To investigate how the multifocal electroretinogram (ERG) is altered in conditions of blocking, light scattering, or distortion of the stimulus that are seen in ocular pathologies. METHODS: A central 40 degree-diameter stimulus pattern consisting of 61 hexagons was presented on a cathode ray tube monitor at a rate of 75 Hz according to the pseudo-random binary M sequence by the Veris computer program. Localized responses corresponding to each hexagon and ERG topographies were displayed on the computer screen. Central scotoma was simulated by blocking the central area of the stimulus, visual field constriction by blocking the outer area of the stimulus, mild cataract by using acrylic filters that caused light scatter, and epiretinal membrane by using a wavy plastic plate that produced metamorphopsia. RESULTS: The responses from the blocked area were nonrecordable whether blockage was central or peripheral; responses from the adjacent unblocked area had a larger amplitude when large areas of the stimulus were blocked. The light scatter that decreased vision from 20/20 to 20/70 did not significantly decrease response amplitudes. Responses from areas in which the stimulus pattern was distorted were minimally affected. CONCLUSIONS: The results show that the system records local ERGs from the macula and outside the macula. It can detect the area where the stimulus is blocked. Moderate light scattering and distortion do not cause loss of local ERG characteristics. PMID- 10672877 TI - Effect of continuous intravenous infusion of carteolol chloride on tissue blood flow in rabbit optic nerve head. AB - PURPOSE: To investigate the effect of an intravenous infusion of carteolol on tissue blood flow in the optic nerve head (ONH) of rabbits. METHODS: Rabbits received either a 3-week topical instillation, or a single intravenous injection (10, 20, 30 microg/kg) or a continuous intravenous injection (2.5, 5, 20, 40, 80 microg/kg per hour) of carteolol. The plasma carteolol level was determined by the gas chromatography negative-ion chemical ionization mass spectrometric method. The ONH blood flow was determined by the hydrogen clearance method. RESULTS: The plasma level of carteolol after a 3-week instillation was 5.55 ng/mL, and a continuous intravenous injection (5 microg/kg per hour) led to approximately the same plasma level. The continuous intravenous infusion of 5 microg/kg per hour of carteolol significantly increased the ONH blood flow compared to the controls from 30 minutes to 2 hours after the beginning of the infusion (n = 10). The mean blood pressure and intraocular pressure (n = 6) were not significantly changed during the continuous intravenous infusion of carteolol. CONCLUSIONS: These results suggest that the plasma carteolol level in rabbits after long-term instillation can increase the ONH blood flow. We conclude that the increase resulted from a reduction in the vascular resistance in the ONH. PMID- 10672878 TI - Alterations of the electroretinogram by intravitreal kainic acid in the rat. AB - PURPOSE: To relate the electrophysiological changes in the retina induced by the excitatory neurotoxin, kainic acid (KA), to its receptor sites in the rat. METHODS: Fifty-five Wistar rats were injected intravitreally with 100, 50, 25, 12.5, 6.25, 3.12, or 1.56 nmol of KA. The electroretinograms (ERGs) including oscillatory potentials (OPs) elicited by a series of increasing intensities were recorded before, and 6 hours, 1 day, 1 week, and 1 month after the injection of KA. RESULTS: After KA injection, the a-waves showed no significant change at all intensity levels (P > .05), but the amplitudes and implicit times of the b-wave were significantly altered. The abolition of the b-wave by KA resulted in a negative response, which decreased progressively with time. The implicit times of the b-wave showed a marked prolongation after injection of 100 nmol of KA (P < .01). The OPs disappeared at the KA dose of 6.25 nmol and higher; doses of 1.56 to 3.12 nmol of KA depressed the Ops. CONCLUSIONS: We conclude that KA altered the above-mentioned ERG components in a dose-dependent manner. These alterations of the ERG can be explained by alterations of neurons in the inner retinal layers. PMID- 10672880 TI - Sicca syndrome and HTLV-I-associated myelopathy/tropical spastic paraparesis. AB - PURPOSE: The objective of this study is to describe the clinical and immunological aspects observed in patients with both human T-cell lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis and ocular dryness. METHODS: In 15 such patients, clinical and biological examinations completed with a biopsy of secondary salivary glands were performed to assess the etiology of the ocular dryness. RESULTS: Histological study of the biopsy specimens indicated that 80% of the patients had grade 3 or grade 4 lesions, according to the Chisholm scale. Polyclonal hypergammaglobulinemia was found in 60% of patients and lymphocytic alveolitis in 80%. Three patients had past medical history of chronic uveitis. CONCLUSIONS: All findings in these patients were compatible with Sjogren's syndrome; however, no immunological disorders characteristic of the syndrome were found. Tests for antinuclear antibodies and rheumatoid factor proved negative in all cases. PMID- 10672879 TI - Ocular surface management in corneal transplantation, a review. AB - PURPOSE: To discuss the importance of ocular surface management in corneal transplantation, especially in limbal transplantation. METHODS: Since the corneal epithelium is not completely recovered after corneal transplantation, meticulous attention should be paid to the ocular surface to prevent infection and rejection, which are the major causes of corneal transplantation failure. Preoperative evaluations of the ocular surface should be carried out, followed by appropriate surgical procedures, depending on the condition of each patient. Vigorous immunosuppressive measures should be taken after surgery. RESULTS: In both case reports presented in this study, each patient underwent successful surgery and his condition was controlled by medication suited to his needs. CONCLUSIONS: For those patients with stem cell deficiency, limbal transplantation, possibly with the use of autologous serum drops, should be considered to reconstruct and maintain the ocular surface. Ocular surface management is necessary for the success of corneal transplantation, especially for limbal transplantation. PMID- 10672881 TI - Long-term follow-up of excimer laser phototherapeutic keratectomy. AB - PURPOSE: To evaluate long-term follow-up results of excimer laser phototherapeutic keratectomy (PTK) in a Japanese population. METHODS: Twenty-six patients (31 eyes) with corneal opacity were treated with excimer laser PTK. Preoperative diagnoses included 16 eyes with band keratopathy, 10 with granular dystrophy, and 5 with corneal scar. Mean postoperative follow-up was 27 months. RESULTS: Corneal opacity was reduced in all patients. At postoperative month 12, best spectacle-corrected visual acuity (BSCVA) improved from the preoperative level in 22 eyes of 28 eyes, did not change in 3 eyes, and declined in 3 eyes. BSCVA at month 24 was better than the preoperative acuity in 17 eyes of 23 eyes, similar in 1 eye, and worse in 5 eyes. Eyes with granular dystrophy showed significantly better BSCVA improvement than those with band keratopathy. A hyperopic shift of +1.0 diopter or more occurred in 14 eyes of 28 eyes at month 12 and in 12 eyes of 23 eyes at month 24. No serious adverse effects were encountered during the 3-year follow-up period. CONCLUSIONS: Excimer laser PTK is a safe and effective procedure for the treatment of Japanese patients with superficial corneal opacity. PMID- 10672882 TI - Therapeutic keratoplasty using preserved corneas from keratoconus eyes. AB - PURPOSE: To report and discuss cases of lamellar keratoplasty using corneas obtained during previous penetrating keratoplasty in keratoconus eyes. METHODS: Corneal buttons were obtained from 7 keratoconus patients and stored in a preserving solution for 7-60 days (average, 32.4 days) before use. The recipient eyes comprised recurrent pterygium 3 eyes, primary pterygium 1 eye, pseudopterygium 1 eye, corneal perforation with iris prolapse due to fungal corneal ulcer 1 eye, and limbal dermoid 1 eye. RESULTS: The recipient eyes ran favorable courses in general. Graft rejection developed in 2 eyes and was successfully treated with topical and systemic corticosteroid. CONCLUSIONS: Preserved corneas from keratoconus eyes were found useful in therapeutic lamellar keratoplasty. By this procedure, the current inadequate supply of donor corneas in eye banks in Japan can be augmented. PMID- 10672883 TI - Strabismus surgery using the intraoperative adjustable suture method under anesthesia with propofol. AB - PURPOSE: To evaluate the feasibility and efficacy of using the intraoperative adjustable suture method with anesthesia induced by intravenously administered propofol for strabismus surgery. METHODS: Seven adult patients (mean age, 29.7 +/ 18.5 years) with different types of strabismus were enrolled in this study. All patients underwent full ophthalmological and general medical examinations before surgery. Surgery was performed after induction of anesthesia using intravenously administered propofol that was titrated to control consciousness. RESULTS: Arousal of consciousness was observed at approximately 2 minutes after discontinuation of the propofol infusion in each case, and the consciousness level was sufficient to allow accurate cover-uncover testing and intraoperative adjustment of sutures. Minor complications of nausea in three patients and vomiting in one patient were noted after surgery. CONCLUSIONS: Strabismus surgery using the adjustable suture method with propofol intravenous anesthesia appears to be safe and useful for the treatment of adult strabismus. PMID- 10672884 TI - Ultrasound biomicroscopy dark room provocative testing: a quantitative method for estimating anterior chamber angle width. AB - PURPOSE: To describe a quantitative method for measuring the iridocorneal angle recess area, and, using this, to evaluate factors associated with appositional angle-closure during dark room provocative testing using ultrasound biomicroscopy (UBM). METHODS: All patients (178 patients, 178 eyes) with clinically narrow angles referred for UBM dark room provocative testing between September 1996 and March 1998 were enrolled in this study. Images of the inferior quadrant of the angle taken under standardized dark and light conditions were analyzed. The angle recess area (ARA) was defined as the triangular area demarcated by the anterior iris surface, corneal endothelium, and a line perpendicular to the corneal endothelium drawn from a point 750 microm anterior to the scleral spur to the iris surface. ARA, and acceleration and gamma-intercept of the linear regression analysis of the ARA were calculated. In the linear regression formula, y = ax + b, the acceleration a describes the rate at which the angle widens from the scleral spur; the y-intercept b describes the distance from the scleral spur to the iris. RESULTS: Under dark conditions, the angles in 99 patients (55.6%) showed evidence of appositional angle-closure during testing. ARA (0.11 +/- 0.04 vs. 0.15 +/- 0.05 mm2, P < .0001, Student t-test), acceleration a (0.22 +/- 0.15 vs. 0.26 +/- 0.17, P = .068), and y-intercept b (66 +/- 46 vs. 92 +/- 47 microm, P = .0003) were smaller in eyes that were occluded. In the eyes that were not occluded, y-intercept b showed no significant difference between light and dark conditions (P = .1, paired t-test), while acceleration a did (P < .0001). In the eyes that were occluded, both decreased significantly under dark conditions (P < .0001). CONCLUSIONS The ARA linear regression formula provides useful quantitative information about angle recess anatomy. The more posterior the iris insertion on the ciliary face, the less likely the provocative test will be positive. PMID- 10672885 TI - Pigmentation on anterior chamber angle in eyes of patients with atopic dermatitis. AB - PURPOSE: To evaluate pigmentation on the anterior chamber angle in patients with atopic dermatitis. METHODS: This study includes 61 patients suffering from atopic dermatitis who visited our hospital between 1991 and 1995. Gonioscopy, cycloscopy, and fundus examinations with a scleral depressor were performed on every patient during the initial visit, and were repeated every 6 months. Pigmentation on the anterior chamber angle was classified according to Scheie. RESULTS: The pigmentation on the anterior chamber angle at the initial visit was evaluated as grade 0 (15 eyes), grade 1 (81 eyes), grade 2 (21 eyes), and grade 3 (5 eyes). Retinal detachment was found in 9 eyes of the 26 eyes with grade 2 or 3 pigmentation, but in only one of the 96 eyes with grade 0 or 1. The pigmentation on the anterior chamber angle correlated significantly with the incidence of retinal detachment (P < .0001). Causative breaks were found in the peripheral retina or ciliary epithelium in all eyes. CONCLUSIONS: Moderate to dense pigmentation on the anterior chamber angle in patients with atopic dermatitis seems to be a sign of breaks in the retina or ciliary epithelium, and the fundus of these patients should be examined carefully for signs of retinal detachment. PMID- 10672886 TI - Prevalence of normal-tension glaucoma and primary open-angle glaucoma in patients with collagen diseases. AB - PURPOSE: To investigate the prevalence of normal-tension glaucoma (NTG) and primary open-angle glaucoma (POAG) in patients with collagen diseases and determine whether an immunocompromised condition is present in a subset of glaucoma patients. METHODS: Three glaucoma specialists prospectively examined patients with collagen diseases. The diagnostic process included applanation tonometry, slit-lamp examination, gonioscopy, direct ophthalmoscopy, and automated static perimetry. Twenty-four-hour intraocular pressure monitoring was done when necessary. Using the results of a population-based survey conducted in Japan, we calculated an expected number of cases of NTG and POAG, and compared these with the actual number of cases. RESULTS: Of the 153 patients with collagen diseases examined, we found 6 patients with NTG and 2 patients with POAG. Of these 8 patients, 2 with progressive systemic sclerosis (PSS), one with NTG, and the other, POAG, had a history of being on systemic steroidal therapy. The prevalence of NTG and POAG was significantly higher in women patients having collagen diseases as compared with normal women (P = .027). CONCLUSION: Women patients with collagen diseases are highly susceptible to NTG and POAG. PMID- 10672887 TI - Vitreous changes after treatment of retinopathy of prematurity. AB - PURPOSE: To investigate the vitreous findings in patients with cicatricial retinopathy of prematurity (ROP) who underwent retinal cryopexy and/or photocoagulation during the acute phase of the disease. METHODS: Vitreous findings were evaluated in 15 patients (29 eyes) with cicatricial ROP by slit lamp biomicroscopy and indirect ophthalmoscopy. RESULTS: The ocular examination revealed that all eyes had extensive vitreous liquefaction that affected a large segment of the vitreous. A great deal of fibrillar condensation of the vitreous was present in membrane-like vitreous fibers that traversed the vitreous cavity to the periphery of the degenerating retina. These vitreous changes were most marked in the areas in which retinal cryopexy and/or photocoagulation had been performed. Despite advanced liquefaction, the posterior cortical vitreous was not separated from the retina in any eyes. CONCLUSION: In eyes with ROP that underwent retinal cryopexy and/or photocoagulation during the acute phase of the disease, the vitreous was abnormal, which may contribute to vitreoretinal traction that eventually leads to retinal breaks and detachment. PMID- 10672888 TI - Importance of fluorescein angiographic study in evaluating early retinal changes in Takayasu disease. AB - PURPOSE: To determine the usefulness of fluorescein angiography in studying Takayasu disease. METHODS: We examined 31 eyes in 16 patients with Takayasu disease using indirect ophthalmoscopy, color photography, and fluorescein angiography. Ophthalmoscopic and fluorescein angiographic findings were compared. RESULTS: Fluorescein angiography revealed no additional retinal changes in 10 eyes that had no retinal vein dilatation as seen by indirect ophthalmoscopy. Seven (33%) of 21 eyes that had dilated retinal veins also had additional abnormal findings, such as microaneurysms, arteriovenous shunts, retinal neovascularization, and avascular areas. Some differences in grading the stages of retinopathy were noted with these newly found retinal changes, as compared with the classifications determined by ophthalmoscopy alone. CONCLUSIONS: In Takayasu disease, studying the fundus of patients with fluorescein angiography is particularly important in correctly classifying the stages of retinopathy when the retinal vein appears dilated in ophthalmoscopic observation. PMID- 10672889 TI - White thread-like retinal arterioles associated with antiphospholipid antibody syndrome. AB - BACKGROUND: Report of 2 patients with antiphospholipid antibody syndrome (APS) who had elevated anti-beta2-glycoprotein I antibodies and showed white thread like retinal arterioles. CASES: A complete ophthalmological examination was conducted on 2 patients who presented with blurred or distorted vision. Fluorescein angiography was used to examine the integrity of the retinal circulatory system. Laboratory blood studies were conducted. OBSERVATIONS: In both patients, some of the major retinal arterioles appeared white and had a thread-like appearance. Fluorescein angiography demonstrated progressive occlusion or stenosis of these major arterioles with extensive insufficiency of the regional capillary bed. Patient 2 had systemic lupus erythematosus and was treated with oral corticosteroid and aspirin. Recanalization occurred during a 3 year follow-up in one of the patients. CONCLUSIONS: APS should be considered in cases of white thread-like retinal arterioles. Occlusion of the retinal arterioles in APS may be progressive and responsible for the chronic hypoxia of the retina. PMID- 10672890 TI - The results of serial dynamic enhanced computed tomography in patients with carotid-cavernous sinus fistulas. AB - PURPOSE: To assess serial dynamic enhanced computed tomography (serial DE-CT) as a diagnostic tool for carotid-cavernous sinus fistula (CCF). METHODS: Serial DE CT was performed in seven patients (ages 31-74) with CCF. Contrast material was injected intravenously at a dose of 60 mL with an injection speed of 4 mL per second. Serial axial images of the cavernous sinus were undertaken every 3 seconds using a helical computed tomography system. This relatively low-risk technique provides direct evidence of the arteriovenous shunt in the cavernous sinus. RESULTS: In early imaging after the injection, enhancement of the cavernous sinus on the side of the CCF was noted at the arterial phase in all patients, whereas early enhancement of the cavernous sinus was not observed on the contralateral uninvolved side. CONCLUSIONS: These findings suggest the usefulness of serial DE-CT as a diagnostic tool for the initial diagnosis of both high- and low-flow CCFs. PMID- 10672891 TI - Iodine supplementation: benefits outweigh risks. AB - In 1990, iodine deficiency affected almost one-third of the world population and was the greatest single cause of preventable brain damage and mental retardation. Following a resolution adopted by the World Summit for Children in 1990. major programmes of iodine supplementation were implemented by the governments of the affected countries with the support of major donors. Iodisation of salt was recognised as the method of choice. Nine years later, by April 1999, 75% of the affected countries had legislation on salt iodisation and 68% of the affected populations had access to iodised salt. The prevalence of iodine deficiency disorders decreased drastically in most countries and the deficiency disappeared completely in some such as Peru. This result constitutes a public heath success unprecedented with a non-infectious disease. However, occasional adverse effects occurred. The principle effect is iodine-induced hyperthyroidism which occurs essentially in older people with autonomous nodular goitres, especially following iodine intake that is too rapid and of too massive an increment. The incidence of the disorder is usually low and reverts spontaneously to the background rate of hyperthyroidism or even below this rate after 1 to 10 years of iodine supplementation. The possible occurrence of iodine-induced thyroiditis in susceptible individuals has not been clearly demonstrated by large epidemiological surveys. Iodine supplementation is followed by an increased prevalence of occult papillary carcinoma of the thyroid discovered at autopsy but the prognosis of thyroid cancer is improved due to a shift towards differentiated forms of thyroid cancer that are diagnosed at earlier stages. Iodine-induced hyperthyroidism and other adverse effects can be almost entirely avoided by adequate and sustained quality control and monitoring of iodine supplementation which should also confirm adequate iodine intake. Available evidence clearly confirms that the benefits of correcting iodine deficiency far outweigh the risks of iodine supplementation. PMID- 10672892 TI - Use of routine healthcare data in safe and cost-effective drug use. AB - Routine healthcare data is becoming widely available, usually as a result of administrative systems. Other related data are also often available, such as biochemistry results, mortality data, and sometimes prescribing data. These records are often linked via a common identification system or by probability matching techniques. These data sources offer many opportunities to undertake research, and where prescription data are recorded and linked, the facility to research the outcome of drug use often exists. There are now a number of research agencies around the world that use these large routine data sources to undertake drug safety and outcome studies. The purpose of this commentary is to describe some of the history behind the development of these systems, illustrate some of their uses with respect to postmarketing drug safety and to other healthcare research objectives. The review then describes the data sources necessary to develop a system that would offer an optimal system to undertake a range of studies, including population drug safety surveillance. There are both positive and negative considerations when using routine data. On the positive side, these data come from 'real life' experiences and not from the clinical trial situation. On the other hand, there are important biases to be aware of such as confounding by indication. On the whole, it is argued that large databases originating from routine healthcare procedures have an important role to play in the cost effective prescription drug use in the postmarketing setting. These systems cannot replace other methods of drug safety evaluation but they do offer an important adjunct to spontaneous reporting systems. PMID- 10672893 TI - Patient-oriented strategies for the prevention of drug interactions. AB - Drug interactions are a common and serious problem arising from polypharmacy. Strategies to reduce the likelihood of the co-prescription of hazardous drug combinations are likely to enhance the quality of care provided for patients requiring polypharmacotherapy. Drugs for which patient-oriented information strategies may decrease the likelihood of drug interactions tend to be those of low therapeutic index. and have interaction potential with other drugs commonly prescribed or available without prescription. PMID- 10672894 TI - Corticosteroid-induced adverse psychiatric effects: incidence, diagnosis and management. AB - Reports of corticosteroid-induced adverse psychiatric effects began to appear in the literature soon after the introduction of these medications in the 1950s. Unfortunately, early studies relied on informal classification and measurement procedures and tended to utilise nonspecific descriptive terminology (such as steroid psychosis'). A growing number of contemporary investigations have begun to address these problems. However, the literature remains surprisingly undeveloped from a pharmacoepidemiological perspective, consisting largely of case reports and case series. The objective of this review is to summarise published data concerning corticosteroid-induced adverse psychiatric effects. A clinical perspective will be adopted since opportunities to minimise the impact of corticosteroid-induced adverse effects tend to present themselves most readily within the sphere of clinical management. Some of the psychiatric adverse effects of corticosteroids are mild, and not necessarily clinically significant. However, several serious psychiatric syndromes can be caused by corticosteroids: substance induced mood disorders (with depressive, manic and mixed features), substance induced psychotic disorders and delirium. While certain clinical groups may be at greater risk of corticosteroid-induced adverse psychiatric effects, corticosteroid-induced psychiatric toxicity is remarkably unpredictable. The literature regarding prevention and treatment of corticosteroid-induced adverse psychiatric effects is poorly developed. As a result, the emphasis of this review is on clinical and epidemiological evidence linking specific adverse effects to corticosteroid medications. However, clinical reports do provide some practical guidance for prevention and treatment, and these are summarised as well. A variety of pharmacological strategies for treatment and prevention have been proposed. Education and support also appear to be important, and perhaps neglected. PMID- 10672895 TI - Utility of acetylcysteine in treating poisonings and adverse drug reactions. AB - As recognition of the role of free radicals and reactive toxins in the pathogenesis of disease, poisoning, and adverse drug reactions has evolved, interest in the use of acetylcysteine as a modulator of these effects has steadily increased in recent years. Acetylcysteine is commonly thought to serve as a glutathione precursor and consequently can increase or sustain intracellular glutathione which scavenges reactive oxygen species caused by toxins or subsequent tissue injury. At least 10 additional mechanisms of action for acetylcysteine have been demonstrated in various laboratory models, but a unifying framework of its actions is still to be proposed. This paper reviews the current experimental and therapeutic status of acetylcysteine for the treatment of poisonings and adverse drug reactions. Of the 45 potential uses of acetylcysteine that were identified for the treatment of poisonings or adverse drug reactions, 14 of the toxic effects have little support for its use while promising results have been demonstrated for 27 toxicities. Currently, treatment of acute paracetamol (acetaminophen) poisoning is the only widely accepted clinical indication for acetylcysteine as a treatment for poisoning or adverse drug reactions. In many clinical situations acetylcysteine is used empirically utilising modifications of dosage regimens employed for paracetamol poisoning. Often it is difficult to determine the benefit of therapy with acetylcysteine owing to the nature of the toxicity being treated, the use of other therapies, the presence of comorbid conditions, and the small number of patients studied. The diverse and positive nature of the investigations suggest that there is considerable promise in acetylcysteine as a research tool and pharmacological agent. PMID- 10672896 TI - Managing the adverse effects of interferon-beta therapy in multiple sclerosis. AB - Interferon-beta is an established therapy in relapsing-remitting multiple sclerosis. Recently, it has also been shown that interferon-beta-1b is effective in secondary progressive multiple sclerosis. However, adverse effects of interferon-beta treatment are common, particularly during the first weeks of treatment, and are a major concern. Flu-like symptoms, injection site reactions and laboratory abnormalities are the most common adverse effects, and may result in reduced compliance or even discontinuation of treatment in a number of patients. Therefore, efforts to minimise these reactions, e.g. appropriate comedication with analgesic/antipyretic drugs, use of correct preparation and injection technique and sometimes modification of the dosage of interferon-beta, are of considerable importance. This article provides an overview of the management of clinically relevant adverse effects related to treatment with interferon-beta, based on a literature review and personal experience. Essential aspects of patient information are also stressed. If these recommendations are followed, adverse effects related to interferon-beta may be substantially reduced in the majority of patients. PMID- 10672897 TI - Incidence and costs of adverse drug reactions during hospitalisation: computerised monitoring versus stimulated spontaneous reporting. AB - OBJECTIVE: To implement a computer-based adverse drug reaction monitoring system and compare its results with those of stimulated spontaneous reporting, and to assess the excess lengths of stay and costs of patients with verified adverse drug reactions. DESIGN: A prospective cohort study was used to assess the efficacy of computer-based monitoring, and case-matching was used to assess excess length of stay and costs. SETTING: This was a study of all patients admitted to a medical ward of a university hospital in Germany between June and December 1997. PATIENTS AND PARTICIPANTS: 379 patients were included, most of whom had infectious, gastrointestinal or liver diseases, or sleep apnoea syndrome. Patients admitted because of adverse drug reactions were excluded. METHODS: All automatically generated laboratory signals and reports were evaluated by a team consisting of a clinical pharmacologist, a clinician and a pharmacist for their likelihood of being an adverse drug reaction. They were classified by severity and causality. For verified adverse drug reactions, control patients with similar primary diagnosis, age, gender and time of admission but without adverse drug reactions were matched to the cases in order to assess the excess length of hospitalisation caused by an adverse drug reaction. RESULTS: Adverse drug reactions were detected in 12% of patients by the computer-based monitoring system and stimulated spontaneous reporting together (46 adverse reactions in 45 patients) during 1718 treatment days. Computer-based monitoring identified adverse drug reactions in 34 cases, and stimulated spontaneous reporting in 17 cases. Only 5 adverse drug reactions were detected by both methods. The relative sensitivity of computer-based monitoring was 74% (relative specificity 75%), and that of stimulated spontaneous reporting was 37% (relative specificity 98%). All 3 serious adverse drug reactions were detected by computer-based monitoring, but only 2 out of the 3 were detected by stimulated spontaneous reporting. The percentage of automatically generated laboratory signals associated with an adverse drug reaction (positive predictive value) was 13%. The mean excess length of stay was 3.5 days per adverse drug reaction. 48% of adverse reactions were predictable and detected solely by computer-based monitoring. Therefore, the potential for savings on this ward from the introduction of computer-based monitoring can be calculated as EUR56 200/year ($US59 600/year) [ 1999 values]. CONCLUSION: Computer monitoring is an effective method for improving the detection of adverse drug reactions in inpatients. The excess length of stay and costs caused by adverse drug reactions are substantial and might be considerably reduced by earlier detection. PMID- 10672898 TI - Selective loss of the transforming growth factor-beta apoptotic signaling pathway in mutant NRP-154 rat prostatic epithelial cells. AB - Retroviral insertional mutagenesis was used to select mutant NRP-154 rat prostate carcinoma cells resistant to transforming growth factor (TGF)-beta-induced cell death. Similar to the parental cells, a mutant clone, M-NRP1, expressed TGF-beta receptors and was still responsive to induction both of direct target genes by TGF-beta and of apoptosis by staurosporine or okadaic acid. In contrast, indicators of cell growth, strongly suppressed by TGF-beta in the parental cells, were unaffected in M-NRP1 cells. M-NRP1 cells overexpress the antiapoptotic protein, Bcl-xL, and show dysregulated expression and localization of a protein related to a novel human septin, ARTS (designation of apoptotic response to TGF beta signals), cloned by homology to an exonic sequence flanked by the viral long terminal repeats in M-NRP1 cells and shown to make cells competent to undergo apoptosis in response to TGF-beta. We propose that ARTS might operate within the same apoptotic pathway as Bcl-xL and that M-NRP1 cells could serve as a useful model for characterization of this pathway. PMID- 10672899 TI - Molecular cloning of a novel retinoic acid-responsive gene, HA1R-62, which is also up-regulated in Hoxa-1-overexpressing cells. AB - Using a PCR-based cDNA subtractive hybridization method (L. Diatchenko et al., Proc. Natl. Acad. Sci. USA, 93: 6025-6030, 1996), we cloned a cDNA fragment of a novel gene that is highly expressed in F9-10; F9-10 is an F9 teratocarcinoma stem cell line that expresses high levels of exogenous Hoxa-1 mRNA and protein in comparison to F9 wild-type stem cells, which do not express endogenous Hoxa-1 mRNA in the absence of retinoic acid (RA). Rapid amplification of cDNA ends was used to clone the full-length cDNA of this gene, designated HA1R-62 (Hoxa1 regulated-62). We have shown that HA1R-62 is also a RA-responsive gene and that it is expressed (mRNA size, approximately 4.3 kb) in adult mouse thymus, lung, kidney, and ovary as well as in 12.5-day mouse embryos. DNA sequence analysis and in vitro translation experiments have shown that HA1R-62 encodes a protein with a molecular mass of approximately 26 kDa. Elucidation of the function of the HA1R 62 gene product will provide new insights into the functions of RA and homeobox genes. PMID- 10672900 TI - Thrombin causes pseudopod detachment via a pathway involving cytosolic phospholipase A2 and 12/15-lipoxygenase products. AB - Thrombin causes rapid pseudopod detachment and shortening in Dunning rat prostatic carcinoma (MAT-Lu) cells. As seen by interference reflection microscopy and by immunofluorescence analysis with antibodies to paxillin and talin, the primary event is disassembly of adhesion sites. Biochemically, thrombin is a potent activator of cytosolic phospholipase A2 and increases eicosanoid production in these cells. The pseudopod effects are blocked by lipoxygenase (but not cyclooxygenase) inhibitors. Arachidonic acid and 12(S) hydroxyeicosatetraenoic acid or 15(S)-hydroxyeicosatetraenoic acid mimic the thrombin effect. We conclude that in certain cancer cells, thrombin is a pseudopod repellent that exerts its effect via a cascade involving cytosolic phospholipase A2, 12/15-lipoxygenase, and 12(S)- and/or 15(S) hydroxyeicosatetraenoic acid. PMID- 10672901 TI - Polyoma virus middle T and small t antigens cooperate to antagonize p53-induced cell cycle arrest and apoptosis. AB - Wild-type p53 triggers two distinct biological responses, cell cycle arrest and apoptosis. Several small DNA tumor viruses encode proteins that bind p53 and thus block the function of p53. This probably reflects the need of these viruses to prevent p53-induced cell cycle arrest and apoptosis to allow viral DNA replication. Unlike SV40 large T, polyoma virus large T does not bind p53, and it is still unclear how polyoma virus blocks p53 function. To address this question, we transfected polyoma virus middle T or small t alone or middle T and small t together into J3D mouse T-lymphoma cells carrying temperature-sensitive p53 (ts p53). Induction of wild-type p53 by temperature shift to 32 degrees C triggered both G1 cell cycle arrest and apoptosis in parental J3D-ts p53 cells. In contrast, J3D-ts p53 cells coexpressing middle T and small t showed only a weak G1 cell cycle arrest response after induction of wild-type p53 at 32 degrees C. Fluorescence-activated cell sorter analysis revealed that nearly half of the middle T-expressing cells, 30% of the small t-expressing cells, and a majority of the cells coexpressing middle T and small t were resistant to p53-induced apoptosis. The phosphatidylinositol 3-kinase inhibitor wortmannin partially abrogated the protective effect of middle T but not small t on p53-induced apoptosis, indicating that middle T prevents p53-induced apoptosis through the phosphatidylinositol 3-kinase signal transduction pathway. Our results thus establish a mechanism for polyoma virus-mediated inhibition of p53 function. PMID- 10672902 TI - Phosphospecific antibodies reveal focal adhesion kinase activation loop phosphorylation in nascent and mature focal adhesions and requirement for the autophosphorylation site. AB - Focal adhesion kinase (FAK) is a key signaling molecule regulating cellular responses to integrin-mediated adhesion. Integrin engagement promotes FAK phosphorylation at multiple sites to achieve full FAK activation. Phosphorylation of FAK Tyr-397 creates a binding site for Src-family kinases, and phosphorylation of FAK Tyr-576/Tyr-577 in the kinase domain activation loop enhances catalytic activity. Using novel phosphospecific antibody reagents, we show that FAK activation loop phosphorylation is significantly elevated in cells expressing activated Src and is an early event following cell adhesion to fibronectin. In both cases, this regulation is largely dependent on Tyr-397. We also show that the FAK activation loop tyrosines are required for maximal Tyr-397 phosphorylation. Finally, immunostaining analyses revealed that tyrosine phosphorylated forms of FAK are present in both newly forming and mature focal adhesions. Our findings support a model for reciprocal activation of FAK and Src family kinases and suggest that FAK/Src signaling may occur during both focal adhesion assembly and turnover. PMID- 10672903 TI - Nonapoptotic cell death associated with S-phase arrest of prostate cancer cells via the peroxisome proliferator-activated receptor gamma ligand, 15-deoxy delta12,14-prostaglandin J2. AB - 15-Deoxy-delta12,14-prostaglandin J2 (15d-PGJ2) is a highly specific activator of the peroxisome proliferator-activated receptor gamma (PPAR-gamma). We investigated the effect of 15d-PGJ2 on three human prostate cancer cell lines, LNCaP, DU145, and PC-3. Western blotting demonstrated that PPAR-gamma1 is expressed predominantly in untreated prostate cancer cells. Treatment with 15d PGJ2 caused an increase in the expression of PPAR-gamma2, whereas PPAR-gamma1 remained at basal levels. PPARs alpha and beta were not detected in these cells. Lack of lipid accumulation, increase in CCAAT/enhancer binding proteins (C/EBPs), or expression of aP2 mRNA indicated that adipocytic differentiation is not induced in these cells by 15d-PGJ2. 15d-PGJ2 and other PPAR-gamma activators induced cell death in all three cell lines at concentrations as low as 2.5 microM (similar to the Kd of PPAR-gamma for this ligand), coinciding with an accumulation of cells in the S-phase of the cell cycle. Activators for PPAR-alpha and beta did not induce cell death. Staining with trypan blue and propidium iodide suggested that, although the plasma membrane appears intact by electron microscopy, disturbances are evident as early as 2 h after treatment. Mitochondrial transmembrane potentials are significantly reduced by 15d-PGJ2 treatment. In addition, treatment with 15d-PGJ2 resulted in cytoplasmic changes, which are indicative of type 2 (autophagic), nonapoptotic programmed cell death. PMID- 10672904 TI - Inhibition of RNA polymerase I transcription in differentiated myeloid leukemia cells by inactivation of selectivity factor 1. AB - Transcription by RNA polymerase I (pol I) regulates the rate of ribosome biogenesis and the biosynthetic potential of the cell; therefore, it plays an important role in the control of cell growth. Differentiation of the human promyelocytic leukemic cell line U937 is accompanied by drastic decreases in pol I transcriptional activity. We have used cell-free extracts prepared from undifferentiated and differentiated U937 cells to investigate the molecular mechanisms responsible for this inhibitory process. Our analysis indicates that the activity of the TATA binding protein (TBP)/TBP-associated factor (TAF) complex selectivity factor 1 (SL1), one of the factors required for accurate and promoter-specific transcription by RNA pol I, is severely repressed in differentiated U937 cells. Moreover, the reduction in SL1 activity is not a consequence of a decrease in SL1, because there is no detectable difference in the abundance of TBP or TAFs before and after U937 cell differentiation. In conclusion, our results indicate that the selectivity factor SL1 is an important target for the regulation of pol I transcription during cell differentiation. PMID- 10672905 TI - Transgenic models as tools for studying the regulation of human renin expression. AB - Transgenic mice and rats have become popular tools to study the regulation of gene expression and the consequences of protein over-production. Over the past decade, numerous transgenic models have been developed to study the mechanisms of human renin gene expression and the participation of the renin-angiotensin system in the development of hypertension. Herein we will provide an overview of what has been learned from the use of transgenic models for studying the human renin gene. PMID- 10672906 TI - High fat maintenance diet attenuates hindbrain neuronal response to CCK. AB - Rats maintained on a high fat diet reduce their food intake less in response to exogenous cholecystokinin (CCK) than rats maintained on a low fat diet. In addition, inhibition of gastric emptying by CCK is markedly attenuated in rats maintained on a high fat diet. Both inhibition of food intake and gastric emptying by CCK are mediated by sensory fibers in the vagus nerve. These fibers terminate on dorsal hindbrain neurons of the nucleus of the solitary tract and area postrema. To determine whether diet-induced changes in the control of feeding and gastric emptying are accompanied by altered vagal sensory responsiveness, we examined dorsal hindbrain expression of Fos-like immunoreactivity (Fos-li) following intraperitoneal CCK injection of rats maintained on high fat or low fat diets. Following CCK, there were numerous Fos li nuclei in the area postrema and in the commissural and medial subnuclei of the nucleus of the solitary tract of rats maintained on a low fat diet. However, Fos li was absent or rare in the brains of rats maintained on a high fat diet. These data suggest that the vagal sensory response to exogenous CCK is reduced in rats maintained on a high fat diet. Our results also are consistent with our previous findings that CCK-induced reduction of food intake and gastric emptying are both attenuated in rats maintained on a high fat diet. In addition our results support the hypothesis that attenuation of CCK-induced inhibition of food intake and gastric emptying may be due to diet-induced diminution of vagal CCK responsiveness. PMID- 10672907 TI - An interaction of opioids and galanin in dorsal horn of the spinal cord in mononeuropathic rats. AB - The present study was performed in rats with experimentally induced mononeuropathy after common sciatic nerve ligation. The hind-paw withdrawal latencies to thermal and mechanical stimulation were increased significantly after intrathecal injection of 3 nmol of galanin. The increased hind-paw response latencies induced by galanin were attenuated by following intrathecal injection of 22 nmol, but not 11 or 2.75 nmol of the opioid receptor antagonist naloxone. Further, the increased hind-paw response latencies induced by galanin were prevented by following intrathecal injection of 10 nmol of mu-opioid receptor antagonist, beta-funaltrexamine (beta-FNA), but not by 10 nmol of delta-opioid receptor antagonist, natrindole or 10 nmol of kappa-opioid receptor antagonist, nor-binaltorphimine (nor-BNI). Intrathecal 10 nmol of beta-FNA alone had no significant effects on the hind-paw withdrawal responses. These results demonstrate the existence of a specific interaction between galanin and opioids in the transmission of presumed nociceptive information in the spinal cord of mononeuropathic rats. This interaction involves the activation of mu-opioid receptor. PMID- 10672908 TI - Release of neuropeptide Y and hemodynamic changes during surgical removal of human pheochromocytomas. AB - This study investigates the release of Neuropeptide Y from eight human pheochromocytomas. Profil immunoreactive Neuropeptide Y (Ir-NPY) levels during the management of surgery were compared with these of norepinephrine (NE) while hemodynamics were monitored. Plasma IrNPY and NE levels increased during tumor manipulation and returned to near normal one hour after operation. However, Ir NPY levels remained high just after tumor resection while NE levels were significantly decreased. At tumor manipulation and just after tumor resection, plasma Ir-NPY levels were correlated with the systemic vascular resistances (SVR) (r = 0.74; P<0.04 and r = 0.86; P<0.006 respectively). No correlation was found either between plasma Ir-NPY and NE levels or between plasma NE levels and SVR. The release of Ir-NPY from tumor tissue, studied by a superfusion method, exhibited a significant correlation with the plasma Ir-NPY concentrations at the time of corresponding tumor resection (r = 0.95; P<0.007). Chromatographic analysis showed that Ir-NPY in plasma and outflow migrate as human NPY (1-36). These results confirmed that in pheochromocytoma, plasma NPY mainly originates from the tumor and argue for an important role of NPY in pheochromocytoma hypertension as indicated by the correlation between the Ir-NPY levels and the SVR. PMID- 10672909 TI - GLP-1-analogues resistant to degradation by dipeptidyl-peptidase IV in vitro. AB - Glucagon-like peptide-1 (GLP-1) stimulates insulin secretion and improves glycemic control in type 2 diabetes. In serum the peptide is degraded by dipeptidyl peptidase IV (DPP IV). The resulting short biological half-time limits the therapeutic use of GLP-1. DPP IV requires an intact alpha-amino-group of the N-terminal histidine of GLP-1 in order to perform its enzymatic activity. Therefore, the following GLP- analogues with alterations in the N-terminal position 1 were synthesized: N-methylated- (N-me-GLP-1), alpha-methylated (alpha me-GLP-1), desamidated- (desamino-GLP-1) and imidazole-lactic-acid substituted GLP-1 (imi-GLP-1). All GLP-1 analogues except alpha-me-GLP-1 were hardly degraded by DPP IV in vitro. The GLP-1 analogues showed receptor affinity and in vitro biological activity comparable to native GLP-1 in RINm5F cells. GLP-1 receptor affinity was highest for imi-GLP-1, followed by alpha-me-GLP-1 and N-me-GLP-1. Only desamino-GLP-1 showed a 15-fold loss of receptor affinity compared to native GLP-1. All analogues stimulated intracellular cAMP production in RINm5F cells in concentrations comparable to GLP-1. N-terminal modifications might therefore be useful in the development of long-acting GLP-1 analogues for type 2 diabetes therapy. PMID- 10672910 TI - Gastric submucosal microdialysis: a method to study gastrin- and food-evoked mobilization of ECL-cell histamine in conscious rats. AB - Rat stomach ECL cells are rich in histamine and chromogranin A-derived peptides, such as pancreastatin. Gastrin causes the parietal cells to secrete acid by flooding them with histamine from the ECL cells. In the past, gastric histamine release has been studied using anaesthetized, surgically manipulated animals or isolated gastric mucosa, glands or ECL cells. We monitored gastric histamine mobilization in intact conscious rats by subjecting them to gastric submucosal microdialysis. A microdialysis probe was implanted into the submucosa of the acid producing part of the stomach (day 1). The rats had access to food and water or were deprived of food (48 h), starting on day 2 after implantation of the probe. On day 4, the rats received food or gastrin (intravenous infusion), and sampling of microdialysate commenced. Samples (flow rate 1.2 microl min(-1)) were collected every 20 or 60 min, and the histamine and pancreastatin concentrations were determined. The serum gastrin concentration was determined in tail vein blood. Exogenous gastrin (4-h infusion) raised microdialysate histamine and pancreastatin dose-dependently. This effect was prevented by gastrin receptor blockade (YM022). Depletion of ECL-cell histamine by alpha-fluoromethylhistidine, an irreversible inhibitor of the histamine-forming enzyme, suppressed the gastrin evoked release of histamine but not that of pancreastatin. Fasting lowered serum gastrin and microdialysate histamine by 50%, while refeeding raised serum gastrin and microdialysate histamine and pancreastatin 3-fold. We conclude that histamine mobilized by gastrin and food intake derives from ECL cells because: 1) Histamine and pancreastatin were released concomitantly, 2) histamine mobilization following gastrin or food intake was prevented by gastrin receptor blockade, and 3) mobilization of histamine (but not pancreastatin) was abolished by alpha fluoromethylhistidine. Hence, gastric submucosal microdialysis allows us to monitor the mobilization of ECL-cell histamine in intact conscious rats under various experimental conditions not previously accessible to study. While gastrin receptor blockade lowered post-prandial release of ECL-cell histamine by about 80%, unilateral vagotomy reduced post-prandial mobilization of ECL-cell histamine by about 50%. Hence, both gastrin and vagal excitation contribute to the post prandial release of ECL-cell histamine. PMID- 10672911 TI - Interaction between brain natriuretic peptide and atrial natriuretic peptide in caecal circular smooth muscle cells. AB - Guinea pig caecal circular smooth muscle cells were used to determine whether brain natriuretic peptide (BNP) can inhibit the contractile response produced by cholecystokinin-octapeptide (CCK-8). In addition, we examined the effect of an inhibitor of cAMP-dependent protein kinase, an inhibitor of particulate or soluble guanylate cyclase, an atrial natriuretic peptide (ANP) antagonist (ANP 1 11), and selective receptor protection on the BNP-induced relaxation of these muscle cells. The effect of BNP on cAMP formation was also examined. BNP inhibited the contractile response produced by CCK-8 in a dose-response manner, with an IC50 value of 8.5 nM, and stimulated the production of cAMP. The inhibitor of cAMP-dependent protein kinase and the inhibitor of soluble guanylate cyclase significantly inhibited the relaxation produced by BNP. In contrast, the inhibitor of particulate guanylate cyclase did not have any significant effect on the relaxation produced by BNP. ANP 1-11 significantly but partially inhibited the relaxation produced by BNP. The muscle cells where CCK-8 and ANP binding sites were protected completely preserved the inhibitory response to ANP, but partially preserved the inhibitory response to BNP. The muscle cells where CCK-8 and BNP binding sites were protected completely preserved the inhibitory response to both ANP and BNP. This study demonstrates that BNP induces relaxation of these muscle cells via both ANP binding sites coupled to soluble guanylate cyclase and distinct BNP binding sites coupled to adenylate cyclase. PMID- 10672912 TI - Introduction: aging and the kidney. PMID- 10672913 TI - The nature of chronic progressive nephropathy in aging rats. AB - Increases in glomerular size and thickening of the glomerular basement membrane are constant features that accompany growth and maturation in animals. Yet, some animals have chronic progressive nephropathy characterized by glomerulosclerosis and tubulointerstitial fibrosis. In these animals, clinically significant reductions in glomerular filtration rate may compromise health, particularly when other renal diseases occur concomitantly. Progressive thickening of the glomerular basement membrane is accompanied by changes in its composition, which may be responsible for changes in podocyte morphology and proteinuria. Within the tubulointerstitium, generalized accumulation of fibronectin and thrombospondin are accompanied by blood vessel proliferation. Fragility of these blood vessels with intermittent bleeding may initiate an inflammatory process that leads to focal areas of tubular atrophy and scarring. The pathogenesis of these lesions is unknown. Genetic background, sex, and environmental factors influence the tempo of progressive sclerosis, although these factors are not primary determinants of this lesion. This review highlights the structural changes that occur in the kidney with aging. Because the lesions are structurally similar, information gleaned from studies of aging animals should be relevant to understanding the loss of renal function that occurs in aging humans. PMID- 10672914 TI - Biology of renal aging in humans. AB - Advancing age is usually accompanied by a decline in glomerular filtration rate and an increased incidence of certain renal and electrolyte disorders. These include an increased susceptibility to acute renal failure, hypo- and hypernatremia, hyperkalemia, and hypertension. This report discusses anatomic and physiological observations related to the aged human kidney and explores the various theories and postulated mechanisms underlying these changes. PMID- 10672915 TI - Hypertension in older adults. AB - Isolated systolic hypertension and combined systolic and diastolic hypertension are clinical problems affecting significant numbers of older adults (>65 years). Preventing the complications of hypertension, including stroke and coronary artery disease, may potentially impact not only an individual's sense of well being, but also their functional status and ability to live independently in the community. Despite the increased absolute risk for cardiovascular events associated with hypertension in older adults compared with younger adults, significant numbers of individuals remain untreated or inadequately treated. There is clinical data to show that treating both isolated systolic hypertension and combined systolic and diastolic hypertension results in a significant reduction in cardiovascular events in older adults. Although there is a growing body of literature on treatment of hypertension in the 60- to 80-year-old, the data on individuals greater than 80 years old is lacking. The challenge becomes to treat hypertension safely in the presence of multiple medications and other diseases. PMID- 10672916 TI - Diabetes in the elderly population. AB - Type 2 diabetes mellitus has emerged as an important condition of older patients in which both microvascular and macrovascular complications are a common cause of morbidity and mortality. In contrast to type 1 diabetes mellitus, this endocrinopathy is clustered in minority populations and has both strong genetic and environmental factors that influence disease manifestation. A number of physiological alterations of glucose metabolism including hepatic overproduction of glucose, and reduced glucose utilization by peripheral tissues as a result of insulin resistance contribute to the development of the metabolic manifestations of this disease. Ultimately, pancreatic failure and reduced insulin secretion lead to hyperglycemia and the diabetic state. Frequently, many of these metabolic manifestations, or what has been termed Syndrome X, antecede the development of overt diabetes by many years. This syndrome is manifest clinically by such cardiovascular risk factors as hypertension, dyslipidemia, and coagulation abnormalities. This abnormal metabolic milieu contributes to the high prevalence of macrovascular complications including coronary artery disease as well as more generalized atherosclerosis. Microvascular complications have only more recently been recognized as an important and frequent complication of type 2 diabetes. Among the elderly and minority populations, this has become the single most important cause of end-stage renal failure that necessitates renal replacement therapies. The outcome for these patients on hemodialysis, the modality most frequently selected, is poor, with the majority of these patients dying of cardiovascular causes. Unfortunately, interventional strategies to reduce or prevent the microvascular and macrovascular complications have only recently received the needed attention and will require considerable effort and resources to improve the clinical outcomes and life expectancies for these patients. PMID- 10672917 TI - Renal replacement therapy in the elderly: medical, ethical, and psychosocial considerations. AB - As patients over the age of 65 become the fastest growing segment of our treated end-stage renal disease (ESRD) population, nephrologists and allied healthcare workers who care for these patients must become well versed in the many issues specific to this group. Elderly patients contribute the greatest fraction to the incidence and prevalence of the United States ESRD population. Their life expectancy is greatly reduced compared with age-matched counterparts from the general population. Cardiac disease is the leading cause of death. Although renal transplantation remains the most successful form of renal replacement therapy, only a small fraction of elderly ESRD patients are transplanted. The renal research community has made great strides in improving patient outcomes on dialysis over the last decade in many areas; however, little attention has been focused on the elderly ESRD patient. The substantial mortality and comorbidity experienced by this population makes their management an ongoing challenge. Many unresolved issues remain for elderly ESRD patients in the timing of dialysis initiation, choice of dialytic therapy, use of renal transplantation, and management of cardiovascular disease. It is anticipated that future research in these areas will identify optimal treatment strategies for elderly ESRD patients starting on dialysis and improve patient outcomes. PMID- 10672918 TI - Ethical issues in aging and renal disease. AB - The incidence of elderly patients reaching end-stage renal disease (ESRD) and requiring renal replacement is increasing. Better medical care is helping patients live longer but, at the same time, is raising ethical questions. Treatment decisions for ESRD patients present a forum for the consideration of ethical questions surrounding the issues of scarce health care resource allocation and the withholding or withdrawal of life-sustaining treatment. As background for the consideration of ethical issues in ESRD patients, the quality of life they experience and what they may expect as death approaches also are discussed. PMID- 10672919 TI - Successful aging and disease prevention. AB - Substantial increases in the relative and absolute number of older persons in our society pose a challenge for biology, social and behavioral science, and medicine. Successful aging is multidimensional, encompassing the avoidance of disease and disability, the maintenance of high physical and cognitive function, and sustained engagement in social and productive activities. Research has identified factors predictive of success in these critical domains. Two additional research domains, resilience and wisdom, are suggested, and a national initiative in health promotion and disease prevention is proposed. PMID- 10672920 TI - Caring for the nursing home resident on dialysis: a search for solutions. AB - The need for nursing home placement of dialysis patients continues to escalate. Creative solutions are necessary to insure placement availability and optimal care of residents on dialysis once admitted. Communication between the nursing home and dialysis unit staff is key to reducing barriers to placement. The National Kidney Foundation of Michigan is currently intervening through the Nursing Home Manual to enhance training and communication. PMID- 10672921 TI - Educating community providers changes beliefs towards caring for the ESRD patient. AB - End-Stage Renal Disease (ESRD) patients often have comorbid conditions necessitating the involvement of multiple professionals from various settings such as dialysis centers, primary care and skilled nursing facilities, group homes, mental health, rehabilitation, and home health care agencies. This provision of services can be fragmented and may be impeded by a lack of knowledge regarding the specific needs of renal patients. "A Caring Community: An Integrated Approach to Meeting the Needs of the Renal Patient" is a multidisciplinary professional education program which has been shown to change the beliefs of community providers regarding ESRD patients. Program attendee survey responses were measured in accordance with a modified Health Belief Model. This report identifies the need and demand for interdisciplinary renal education and the potential benefits of such a community-wide intervention and makes recommendations for future research. PMID- 10672922 TI - Genetically engineered transgenic plants with the domain 1 sequence of tobacco mosaic virus 126 kDa protein gene are completely resistant to viral infection. AB - In many plant RNA viruses, Domains 1, 2 and 3 are conserved in replicase proteins. In order to examine the interference of viral replication by the Domain 1 sequence, we generated transgenic plants transformed with DNA corresponding to the Domain 1 sequence of the TMV 126 kDa protein. This DNA sequence includes the TMV RNA from nucleotides 1 to 2,149, which comprises both the 5'-untranslated and methyl transferase region. The transgenic plants obtained showed complete resistance to TMV infection. The presence of the Domain 1 sequence in the plants completely prevented local necrosis in Nicotiana tabacum cv. Xanthi nc, and any systemic development of symptoms in Nicotiana tabacum Xanthi upon TMV inoculation. Most transgenic plants sustained the conferred resistance even under TMV inoculum concentrations up to as high as 1,000 microg/ml. To detect any accumulation of TMV coat protein or viral RNA in infected transgenic plants, immunochemical tests and Northern blot analyses were carried out. Neither viral RNA or coat protein was detectable in the systemic leaves of the completely resistant transgenic plants, whereas they were accumulated in large quantities in all of the control plants. Because of the conservation of Domain 1 in many plant RNA viruses, the acquisition of resistance to virus infection using the Domain 1 sequence appears to be a very effective strategy for breeding of viral resistant plants. PMID- 10672923 TI - Ectopic expression of tobacco MADS genes modulates flowering time and plant architecture. AB - MADS genes encode regulatory factors that are involved in various developmental steps of the plant life cycle. During flower development, they regulate the early step of specifying floral meristem identity as well as the later step of determining the fate of floral organ primordia. Here, we report the isolation of two cDNA clones, NsMADS2 and NsMADS3, from a long-day tobacco species, Nicotiana sylvestris, which encode MADS domain-containing proteins. The NsMADS2 amino acid sequence showed 66% identity to SQUA and 64% to AP1. NsMADS3 showed a high degree of amino acid identity with FBP2 (98%), DEFH 72 (89%), DEFH 200 (88%), and AGL9 (76%). RNA blot analyses of NsMADS2 and NsMADS3 revealed that both transcripts were present in floral organs, but not in vegetative organs such as the leaf, root, stem, and 10 d old seedlings. The NsMADS2 transcript was localized in all four whorls and the NsMADS3 transcript was restricted in the three inner whorls of floral organs. The ectopic expression of NsMADS2 using the CaMV 35S promoter caused early flowering and lengthened internode length in transgenic tobacco plants. The ectopic expression of NsMADS3 also caused early flowering phenotype in transgenic tobacco plants, but the plants exhibited reduced apical dominance. Possible implications of these results in relation to the functions of NsMADS2 and NsMADS3 are discussed. PMID- 10672924 TI - Generation of self-antigen reactive, anti-urocortin specific antibodies by immunization of recombinantly expressed urocortin fusion proteins. AB - Urocortin is a recently described 40-meric neuropeptide, which was originally detected in the rat mid-brain and is believed to play a key role in response to stress situations. While its function in the central nervous system is rather well established, the biological role in the periphery is still to be determined. To investigate its distribution and effect on peripheral cells and tissues, in the present study, urocortin was recombinantly expressed and specific antibodies were generated. So far, the immunological detection of urocortin in the rat was largely dependent on antisera generated in rabbits. However, the polyclonal nature of the serum and the remote species origin tend to show cross-reactivities and higher backgrounds. On the other hand, generation of mouse antibodies to rat urocortin was hampered since mouse and rat urocortin sequences are identical, and such antibodies would represent auto-reactive antibodies. Despite such restrictions, the immunization with a combination of various recombinantly expressed urocortin fusion proteins resulted in the successful generation of mouse antiurocortin antisera, whose specificities were confirmed by ELISA and Western blot analysis. To produce the recombinant proteins for immunization, a cDNA encoding the mature urocortin sequence was cloned and expressed in fusion either with the glutathione-S-transferase, the maltose-binding protein, thioredoxin, or a 6X His tag. Depending on the expression system, the solubility and yield of the recombinant proteins greatly varied. Together with the newly generated antibodies, these recombinantly expressed urocortin proteins will serve as valuable tools in further investigations of the biological function of urocortin. PMID- 10672925 TI - Inhibition of trifluoperazine-induced DNA fragmentation by cyclic AMP mediated signaling. AB - Trifluoperazine (TFP), a phenothiazine antipsychotic agent with calmodulin antagonist property, induces DNA fragmentation in a dose- and time-dependent manner in PC12 cells. Various agents affecting calcium mediated intracellular signal transduction such as calcium chelators, calcium ionopores, inhibitors of phospholipase C, and activators/inhibitors of protein kinase C did not block TFP induced DNA fragmentation. Some of these agents themselves induced DNA fragmentation in the conditions under which they were examined. However, cholera toxin (selective Gs activator), forskolin (adenylate cyclase activator) or dibutyryl cyclic AMP (cyclic AMP analogue) inhibited TFP-induced DNA fragmentation in a dose-dependent manner. These results suggest that it is not the calcium but the Gs and adenylate cyclase pathways that play an important role in TFP-induced DNA fragmentation in PC12 cells. PMID- 10672926 TI - Cymbidium mosaic virus coat protein gene in antisense confers resistance to transgenic Nicotiana occidentalis. AB - The nucleotide sequence of the 3'-terminal region of the Korean isolate of cymbidium mosaic virus (CyMV-Ca) from a naturally infected cattleya was determined. The sequence contains an open reading frame (ORF) coding for the viral coat protein (CP) at the 3'-end and three other ORFs (triple gene block or movement protein) of CyMV. The CP gene encodes a polypeptide chain of 220 amino acids with a molecular mass of 23,760 Da. The deduced CP sequence showed a strong homology with those of two CyMVs reported. A construct of the CyMV-Ca CP gene in the antisense orientation in the plant expression vector pMBP1 was transferred via Agrobacterium tumefaciens-mediated transformation into Nicotiana occidentalis which is a propagation host of CyMV. The T1 progeny of the transgenic plants were inoculated with CyMV and found to be highly resistant to CyMV infection. PMID- 10672927 TI - Peroral immunization of microencapsulated human VP8 in combination with cholera toxin induces intestinal antibody responses. AB - To develop an orally delivered subunit vaccine for rotavirus infection, a trypsin cleavage product of VP4, recombinant VP8*, was expressed in Escherichia coli. The recombinant VP8* (rVP8*), purified by affinity chromatography, was reactive against human rotavirus positive serum in Western-blot analysis. To further evaluate the immunogenicity of the oral-delivered rVP8*, it was encapsulated with alginate-microsphere and administered in combination with cholera toxin (CT) as a mucosal adjuvant perorally into mice. The ELISPOT assay showed that the number of rVP8*-specific IgG1 antibody secreting cells increased about 3-fold and about 2 fold in spleen and Peyer's patch, respectively as compared to non-immune mice. In addition, the number of rVP8*-specific IgA antibody secreting cells increased about 2-fold in Peyer's patch. Finally, rVP8*-specific IgA antibody response was significantly enhanced in the intestinal fluids from the mice immunized perorally with encapsulated rVP8* and CT. Taken together, these results indicate that rVP8* possessed proper immunogenicity and it would be potentially useful as a subunit vaccine against rotavirus-associated disease through peroral immunization. PMID- 10672928 TI - TCR internalization induced by peptide/MHC ligands requires the transmembrane domains of alphabeta chains of TCR, but not the expression of CD8 and Thy-1 molecules. AB - T-cell receptor (TCR) internalization occurs via TCR recognition of the peptide/MHC molecule complex on antigen presenting cell (APC). In this study, the requirements for inducing the internalization of TCR molecules on Ld major histocompatibility complex (MHC) class I-restricted T-cells were investigated with 2C cytotoxic T-lymphocyte (CTL) clones with defined peptides as the antigen. To evaluate the function of the transmembrane region of TCR alphabeta chains in TCR internalization, we generated T-cell transfectants expressing the wild type and glycosylphosphatidyl inositol (GPI)-linked form of 2C TCR. Among all peptides forming proper ligands to 2C TCR, only the Qp2Ca peptide induced TCR internalization, which was known to have the highest affinity to both Ld MHC class I molecules and TCR in association with Ld molecules. Such TCR internalization was not observed in cells expressing the GPI-linked form of 2C TCR. Furthermore, the expression of CD8 coreceptor and Thy-1 accessory molecules were both not required for Qp2Ca-induced TCR internalization, and these molecules did not accompany TCR internalization. Altogether, these results suggest that TCR internalization on CTL is not a prerequisite for CTL function. PMID- 10672929 TI - Isolation and characterization of mitochondrial manganese superoxide dismutase (MnSOD) from Capsicum annuum L. AB - A cDNA clone for a mitochondrial manganese superoxide dismutase (MnSOD) was isolated and characterized from red pepper (Capsicum annuum L.). The clone consisted of 941 bp containing one open reading frame (ORF) of 687 bp, 34 bp/220 bp of 5'/3'-untranslated region. Amino acid sequence of the ORF showed the highest homology (86%) with that of Nicotiana plumbaginifolia. It encodes for a polypeptide of 228 amino acids with a molecular mass of 25.5 kDa and a pI value of 8.39. Genomic Southern hybridization suggested that more than one copy are present. Northern hybridization showed that the MnSOD transcript was more abundant in stems than in leaves and roots. When seedlings were treated with arsenate (0.1-10 mM), the MnSOD transcript level increased slightly at 0.1 mM and then dropped, while the Cu/ZnSOD transcript level increased at 1 mM, and also dropped at higher concentrations. PMID- 10672930 TI - The SH2-SH2-SH3 domain of phospholipase C-gamma1 directly binds to translational elongation factor-1alpha. AB - Phospholipase C-gamma1 (PLC-gamma1) is a lipase that hydrolyzes PIP2 to generate two second messengers, IP3 and DAG. By using the yeast two-hybrid system, we identified the translational elongation factor-1alpha (EF-1alpha) as a binding protein of PLC-gamma1 from the human B-lymphocyte library. Direct interaction between EF-1alpha and PLC-gamma1 was confirmed by the in vitro binding experiment using purified PLC-gamma1. Furthermore, from the in vitro binding experiment, we could demonstrate that the carboxyl terminal region of EF-1alpha is involved in the interaction with PLC-gamma1, and that both SH2 and SH3 domains of PLC-gamma1 are required for the interaction with EF-1alpha. In vivo interaction between EF 1alpha and PLC-gamma1 was confirmed by the immunoprecipitation experiment using anti-EF-1alpha antibody. The interaction between EF-1alpha and PLC-gamma1 was enhanced by EGF-treatment. Taken together, we suggest that EF-1alpha might play a role in PLC-gamma1-mediated signal transduction. PMID- 10672931 TI - Genomic organization and sequence of the mud loach (Misgurnus mizolepis) growth hormone gene: a comparative analysis of teleost growth hormone genes. AB - The mud loach (Misgurnus mizolepis) growth hormone (GH) gene was cloned and a comparative analysis on its genomic organization was performed. Based on Southern analysis using various kinds of restriction endonucleases, the GH gene proved to exist as a single-copy gene in the mud loach. The complete nucleotide sequences of a 5.1 kb SacI/EcoRI genomic fragment containing the mud loach GH gene and its 5' flanking sequences as well as a mud loach GH cDNA obtained by rapid amplification of a reverse transcriptase-PCR have been determined. The GH gene spans 2.0 kb from the start codon to the polyadenylation signal, and contains five exons and four introns similar to those of carps and mammals. The evolutionary relation of the mud loach GH gene, inferred by comparative analyses of gene structures and sequences in each exon and intron of representative teleost GH genes, reflects the major phylogenetic groupings of teleost. PMID- 10672932 TI - Effects of dopamine and melatonin on the regulation of the PIT-1 isotype, placental growth hormone and lactogen gene expressions in the rat placenta. AB - Rat placenta produces several members of the placental prolactin-growth hormone (PRL-GH), including placental lactogen (PL) and placental prolactin like protein (PLP), during pregnancy. It is important to study placental local regulators that control the expression of PRL-GH genes. We have previously reported that dopamine (DA) can regulate Pit-1 and PL-II gene expressions. In this study we aimed to investigate the local expression of melatonin receptor 1a (Mel1a) and the effects of DA and melatonin on the expressions of PL-Iv, PL-II, PLP-C genes and Pit-1 gene that are involved in the expression of PRL-GH genes in the rat pituitary and placenta. According to the Northern blot analysis, DA receptor 2 (D2) was expressed in the rat placenta. We also report on the local expression of Mel1a in the rat placenta for the first time. Injected DA agonist, bromocriptine (in vivo) decreased PL-Iv, PLP-C and Pit-1 mRNA levels in the rat placenta. The melatonin agonist, chloromelatonin in culture media also decreased the levels of PL-Iv, PL II and PLP-C mRNA. However, melatonin does not affect the Pit-1 mRNA level. These data suggest that D2 and Mel1a may control the expression of PRL-GH genes in the rat placenta and its response to the extracellular changes of DA and melatonin secreted from the maternal organ. However, Pit-1 may not be involved in the Mel1a induced inhibition of PRL-GH gene expressions in the rat placenta. PMID- 10672933 TI - Kinase-dependent change in the conformation of the leukocyte NADPH oxidase subunit p47phox. AB - The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of O2- from oxygen using NADPH as the electron donor. Dormant in resting neutrophils, the enzyme acquires catalytic activity when the cells are exposed to appropriate stimuli. During activation, the cytosolic oxidase components p47phox and p67phox migrate to the plasma membrane, where they associate with cytochrome b558, a membrane-integrated flavohemoprotein, to assemble the active oxidase. In whole cells and under certain circumstances in the cell-free system, the phosphorylation of p47phox mediates the activation process. It has been proposed that conformational changes in the protein structure of cytosolic factor p47phox may be an important part of the activation mechanism. The total protein steady-state intrinsic fluorescence (an emission maximum of 338 nm) exhibited by the tryptophan residues of p47phox was substantially decreased, reflecting on the conformational change that occurs when p47phox was phosphorylated with protein kinase C. We show here that the phosphorylation of p47phox by protein kinase A or mitogen-activated protein kinase, however, had little effect on the intrinsic fluorescence of p47phox. In addition, the present experiments indicate that in the mutant p47phoxS379A, only the single S-->A mutation appears to be a major importance for the function of p47phox, which is able to undergo the change in conformation that takes place when p47phox is phosphorylated by protein kinase C. PMID- 10672934 TI - Development of new adherent mutant from human myeloma-derived cell line: in vitro model of anaplastic transformation of myeloma. AB - Anaplastic myeloma is a rare but distinct, biologically aggressive variant of myeloma which usually results from dedifferentiation or anaplastic transformation of the myeloma cells. The molecular mechanisms that determine the biologic behavior of anaplastic myeloma and effective treatment modalities have not been well known due to lack of in vitro models. In the present study, we have developed an anaplastically transformed mutant from a human myeloma-derived cell line. In the process of long-term culture of the myeloma-derived IM-9 cell line in low serum and nutrient conditions, an adherent mutant line was developed and named IM-9/AD. This mutant cell line displayed several characteristics resembling anaplastic myeloma such as: 1, large cells with large vesicular nucleus and prominent nucleolus, multinuclearity and high mitotic figures; 2, loss of leukocyte-associated antigens; and 3, higher tumorigenecity in scid mice than its parental cell line. This newly developed mutant cell line may serve as a readily available in vitro model to investigate the biology of anaplastic myeloma. PMID- 10672935 TI - LFA-1- and ICAM-1-dependent homotypic aggregation of human thymocytes induced by JL1 engagement. AB - Cell-cell adhesion is essential for the appropriate immune response, differentiation, and migration of lymphocytes. This important physiological event is reflected in vitro by homotypic cell aggregation. We have previously reported that a 120 kDa cell surface glycoprotein, JL1, is a unique protein specifically expressed by immature double positive (DP) human thymocytes which are in the process of positive and negative selections through the interaction between thymocyte and antigen-presenting cells (APCs). The function of the JL1 molecule, however, is yet to be identified. We show here that anti-JL1 monoclonal antibody (mAb) induced the homotypic aggregation of human thymocytes in a temperature- and Mg2+-dependent manner. It required an intact cytoskeleton and the interaction between leucocyte function associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) since it was blocked by cytochalasin B and D, and mAb against LFA-1 and ICAM-1 which are known to be involved in the aggregation of thymocytes. Translocation of phosphatidylserine (PtdSer) through the cell membrane was not detected, implying that the molecular mechanism of JL-1-induced homotypic aggregation is different from that of CD99-induced homotypic aggregation. In summary, JL1 is a cell surface molecule that induces homotypic adhesion mediated by the LFA-1 and ICAM-1 interaction and cytoskeletal reorganization. These findings suggest that JL1 may be an important regulator of thymocyte development and thymocyte-APC interaction. PMID- 10672936 TI - Cloning, nucleotide sequence and expression of thioltransferase (glutaredoxin) cDNA from Schizosaccharomyces pombe. AB - Thioltransferase (TTase), also known as glutaredoxin (Grx), is an enzyme that catalyzes the reduction of a variety of disulfide compounds, including protein disulfides, in the presence of reduced glutathione. TTase acts as a cofactor for various enzymes such as ribonucleotide reductase. We previously purified a TTase from Schizosaccharomyces pombe and its molecular size was determined. In the present study, a cDNA coding TTase was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization, which was constructed in a plasmid vector pGAD GH, and its corresponding insert was confirmed by Southern hybridization. The nucleotide sequence of the 375 bp long cDNA clone reveals an open reading frame, which encodes a protein of 101 amino acids. The coding region of the original clone was transferred after the lac promoter of pUC13 vector for expression in E. coli, and simultaneously, a suitable Shine-Dalgarno (SD) sequence was added in front of the coding region by PCR. The two primers used for PCR also separately contained BamHI and HindIII restriction sites. The E. coli strain (A434) harboring the pUC13 derivative pKU10 showed a 17.3-fold increase in TTase activity compared to the strain with only the vector plasmid. PMID- 10672937 TI - An element with palindromic structure is required for the expression of TBP (TATA box-binding protein) gene in Drosophila melanogaster. AB - Previously we showed that the 5'-flanking regions between -261 and -207 of the Drosophila melanogaster TBP (TATA box binding protein) gene is important for its expression. We further made serial deletion mutants in this region and analyzed their promoter activities using the transient transfection assay. We found that the 16 bp deletion from -261 to -245 greatly reduces the promoter activity of the Drosophila TBP gene. The 16 bp DNA element contains half of a 11 bp long palindromic sequence, CTTTT-GAAAAG. Disruption of the palindromic sequence by site-directed mutagenesis severely affected promoter activity. In addition, the electrophoretic mobility shift assay showed that the oligonucleotide containing the palindromic sequence can make specific DNA/protein complexes when it was mixed with the Drosophila nuclear extract, suggesting that it interacts with nuclear protein(s). Our data suggest that the palindromic sequence has a critical role in the expression of the Drosophila TBP gene. PMID- 10672938 TI - Dephasing of spin echoes by multiple heteronuclear dipolar interactions in rotational echo double resonance NMR experiments. AB - The application of rotational echo double resonance (REDOR) nuclear magnetic resonance (NMR) for accurate distance measurements has thus far been largely restricted to isolated heteronuclear two-spin systems. In the present paper, the informational content of REDOR curves is explored for systems characterized by multi-spin interactions. To this end, numerical REDOR simulations are presented for cases in which single observe spins S are dipolarly coupled to groups of spins I in distinct geometries. To develop the utility of REDOR for characterizing dipolar couplings in unknown and/or ill-defined geometries, the validity ranges and systematic errors of certain analytical approximations are studied. In the limit of short dipolar evolution times where 0 < deltaS/S0 < or = 0.2 to 0.3, the REDOR difference signal intensity increases approximately proportional to the square of the dipolar evolution time. Here, the curvature depends simply on the second moment M2 characterizing the overall strength of the heterodipolar coupling, irrespective of specific molecular geometries. Fitting experimental REDOR data in this manner produces slight systematic underestimates of M2. However, these errors tend to be counterbalanced by additional systematic errors made by neglecting weak couplings to more remote spins and distribution effects caused by disorder. Based on these findings, the results suggest a convenient method of obtaining site-resolved second moment information in disordered materials. PMID- 10672939 TI - Solid-state single and triple-quantum 93Nb MAS NMR studies of ferroelectric Pb(Mg1/3Nb2/3)O3 and a related pyrochlore. AB - Pb(Mg1/3Nb2/3)O3 (PMN), a well-known relaxor ferroelectric material, and a related pyrochlore phase have been studied by single- and triple-quantum 93Nb MAS NMR spectroscopy. The assignment of the NMR resonances has been attempted. PMID- 10672940 TI - 71Ga NMR of reference GaIV, GaV, and GaVI compounds by MAS and QPASS, extension of gallium/aluminum NMR parameter correlation. AB - We report new measurements of NMR parameters for 71Ga in gallium bearing oxide reference compounds, ranging from perfectly ordered systems to disordered crystalline structures and their aluminate counterparts. Static, MAS, and QPASS spectra are obtained at magnetic fields ranging from 7.0 to 18.8 T. With these results we enhance the previously established correlation between isotropic chemical shifts of 71Ga and 27Al and propose a correlation between gallium and aluminum electric field gradients (EFG). This correlation shows that the EFG at 71Ga sites are generally three times greater than those at equivalent 27Al sites. PMID- 10672941 TI - Strategies for extracting NMR parameters from 23Na MAS, DOR and MQMAS spectra. A case study for Na4P2O7. AB - The 23Na magic-angle spinning (MAS), double rotation (DOR) and multiple-quantum magic-angle spinning (MQMAS) NMR spectra of anhydrous sodium pyrophosphate, Na4P2O7, measured at five different Larmor frequencies (nuL) ranging from 105.8 MHz (corresponding to 400 MHz 1H frequency) to 211.6 MHz (800 MHz) are analysed and the complete set of NMR parameters (C(qcc), etaQ and delta(iso)) of the four crystallographically inequivalent sodium sites were determined with high accuracy. Different approaches of spectra evaluation are discussed and their results are compared. The most reliable results are obtained from a combined evaluation of five DOR and three MQMAS spectra but also from two DOR and one MAS spectra or even from a single MQMAS spectrum all data can be derived. It is shown that Na4P2O7 may serve as a useful reference material for experimental set-up and reliability tests of the various NMR experiments. PMID- 10672942 TI - Observation of hydroxyl groups by 17O solid-state multiple quantum MAS NMR in sol gel-produced silica. AB - 17O NMR parameters (CQ, eta, delta(iso) and T1) are reported for both Si-O-Si and Si-OH fragments within a silica gel. The Si-OH units have a wide spread of parameters but are typically characterised by a very short T1 (approximately 0.1 ms) and CQ < 200 kHz. These observations have extremely important implications for the quantification of such units in these gels and related glassy materials by 17O NMR. In light of these observations, the 17O NMR experiments have been optimised and a distinct resonance from the OH group is observed in 1D static and magic angle spinning (MAS) NMR measurements as well in the multiple quantum (MQ) experiment. PMID- 10672943 TI - The influence of a low protein diet in idiopathic hypercalciuria. AB - A group of 42 hypercalciuric patients (24 males and 18 females) aged 23 to 61 years (mean 45.57+/-12.27) with recurrent stone disease was studied. We applied for a period of 10 days a normocalcium, moderately low protein diet. We found statistically significant variations of azotaemia, venous pH, and vitamin D 1-25. In the 24 h urine collection we found a statistically significant decrease of nitrogen, uric acid, sodium, chloride, calcium, phosphates, oxalate, and hydroxyproline. In conclusion, the present study suggests that the restricted consumption of animal protein can produce a limited effect in urinary biochemistry. The actual efficiency of this dietary restriction on stone formation remains to be evaluated by a prospective long-term study of a larger population. Whether this has an effect on the future incidence of stone formation has to be further verified. PMID- 10672945 TI - Pneumatic lithotriptor--a useful tool for challenging renal stone surgery. AB - Open surgical measures may be undertaken in the treatment of some complex calculi and manoeuvres such as extended pyelolithotomy or nephrolithotomy may be necessary. In an attempt to improve surgical results with less morbidity and maximum ease, we used the Swiss Lithoclast to disintegrate large stones presenting as hard cases. Five patients with renal pelvic stones associated with calyceal stones in two and two further cases with staghorn stones were treated surgically using the pneumatic lithotriptor probe with less dissection and without nephrotomy. Only one of the patients with staghorn stones had residual fragments which were then treated with ESWL. We believe that when open surgical treatment is considered in hard cases, especially in those with small renal pelvises, the best results may be achieved with minimum surgical intervention if it is combined with pneumatic lithotripsy. PMID- 10672944 TI - Mermaid and Potter's syndrome occurring simultaneously. AB - We herein report a case of a female embryo who died in utero and at autopsy she was found to have bilateral renal agenesis with the extrarenal manifestations of Potter's syndrome together with mermaid syndrome which is a rare combination. From all the anomalies of the upper urinary tract bilateral renal agenesis seems to have a cardinal role in the survival of the embryo afflicted with the spectrum of associated anomalies. PMID- 10672946 TI - Arteriovenous shunting in a giant renal angiomyolipoma. A rare condition. AB - We report a case of a 33-year-old woman with tuberous sclerosis and bilateral angiomyolipomas. She suffered from acute left flank pain due to retroperitoneal haemorrhage. During renal arteriography an arteriovenous shunting was found in the left tumour. Angiomyolipoma is a rare cause of angiographically demonstrable arteriovenous shunting. PMID- 10672947 TI - Multicentricity in renal cell carcinoma. AB - OBJECTIVE: To find the incidence of multicentric renal cell carcinoma and its possible relationship to the other clinical and pathologic findings. METHODS: A total of 40 patients with renal cell carcinoma underwent radical nephrectomy between March 1994 and January 1996 at Hacettepe University, School of Medicine, Department of Urology. All of the materials were examined grossly and histologically by the same pathologist. RESULTS: Among 40 kidneys 4 had satellite carcinoma (10%), 3 of them had been shown by preoperative imaging techniques, 1 was found histopathologically. CONCLUSION: If preoperative imaging techniques do not show additional lesion in the kidney besides the small early stage primary in incidentally discovered patients, the incidence of satellite renal cell carcinoma is low enough to justify nephron sparing surgery. PMID- 10672948 TI - Ureteric stone extraction with retrograde irrigational balloon catheter. PMID- 10672949 TI - Retroperitoneal malignant fibrous histiocytoma. AB - Authors review the case history and follow-up of a rare malignant fibrous histiocytoma patient, based on the relevant literary data. The tumour filled the retroperitoneum on the right side, in front of the right kidney. Intravenous urography and computer tomography revealed a 10 x 15 cm sized mass, suspect of being a kidney tumour. Upon surgery, the tumour was found to be a retroperitoneal malignant fibrous histiocytoma. In connection with the case, a brief review is given of the storiform type of malignant fibrous histiocytoma, regarding its aetiological, clinical and pathological aspects, the difficulties in diagnosis, as well as the therapeutic possibilities. Authors regard their case worthy of publication because of the retroperitoneal location and significant size of the tumour, and because of the unproven diagnosis prior to surgery. Even after 4 years the patient is symptom- and complaint-free, and CT has revealed no metastases. PMID- 10672950 TI - The use of polytetrafluoroethylene (Gore-Tex) grafts in reconstruction of the urinary bladder. AB - The safety and histopathologic effects of polytetrafluoroethylene (Gore-Tex) grafts in reconstruction of the urinary bladder were examined. Following partial excision of the bladder Gore-Tex was placed, and the rats were sacrificed at days 7, 14 and 30. Gore-Tex did not cause urine infection, and there was no peritonitis or sepsis in any of the rats. Inflammation around the Gore-Tex diminished after four weeks. Some mononuclear cells and exudate were observed on the inner surface of the Gore-Tex. There was no inflammation or fibrosis in the mucosa and muscular layers of the remaining bladder. In this study the Gore-Tex graft was found to be an infection resistant, urine impermeable material, with no adverse effects on the urinary bladder. Gore-Tex is suggested as a safe material for the reconstruction of the urinary bladder. PMID- 10672951 TI - Vesicovaginal and ureterovaginal fistulas: a review of 39 cases. AB - The aim of this retrospective study was to present our experience in the treatment modalities of patients with vesicovaginal and ureterovaginal fistula. Between 1987 and 1997, 39 patients were diagnosed and operated on for vesicovaginal and ureterovaginal fistula. Of these patients, 31 had vesicovaginal, 7 had ureterovaginal and 1 had both vesicovaginal and ureterovaginal fistula. The ureterovaginal fistulas were repaired by simple ureteroneocystostomy with 100% success. Vesicovaginal and urethrovaginal fistulas were repaired transvaginally in 7 cases, transabdominally in 23 cases and endoscopically in two cases with 77% success at first attempt and 92% success with several attempts. The successful repair of urinary tract fistulas can be achieved in the majority of cases by adhering to the basic surgical principles. The optimum approach is that which works best in the surgeon's hands. PMID- 10672952 TI - Clinical significance of nuclear p53 protein accumulation in bladder cancer. AB - OBJECTIVES: To investigate the correlation of nuclear p53 accumulation with disease outcome in a cohort of patients with transitional cell carcinoma of the bladder. METHODS: A total of 90 patients (11 female, 79 male) with transitional cell carcinoma of the bladder were included in this study. Tumour samples from the primary tumour were analysed by immunohistochemistry for nuclear accumulation of p53 protein. Outcome of each patient was recorded and investigated for a possible relation with p53 status. RESULTS: Nuclear p53 deposition was determined in 22 specimens. The nuclear p53 deposition was seen in less than 20% of the nuclei examined in 13 and more than 20% in 9 cases. No stromal staining was observed. Nuclear p53 deposition was present in 15.2% (7/46) of grade 2 tumours, and 34% (15/44) of grade 3 tumours (p=0.037). Stage distribution revealed 15.5% (5/33) positivity in stage pTa, 25.8% (8/31) in pT1 and 34% (9/26) in stage pT2-3 tumours. Tumours with p53 nuclear accumulation had a higher rate of recurrence and progression and shorter survival. CONCLUSION: Results of the current study indicate p53 as an important factor in determination of biological behaviour of bladder cancer. PMID- 10672953 TI - Nitric oxide based influence of nitrates on micturition in patients with benign prostatic hyperplasia. AB - BACKGROUND: Nitric oxide (NO) is involved in the physiologic regulation of smooth muscle relaxation in the prostate. Organic nitrates act as NO donors. In this prospective open study we prove the influence of orally given nitrates on micturition. METHODS: Thirty-two patients underwent a urological medical check-up prior to starting nitrate medication for cardiovascular disease. We examined peak flow rates, residual urine, IPS-score, PSA level and prostate volume. Exact inclusion and exclusion criteria were defined. Fifteen patients suffered from obstructive symptoms, 17 patients reported no subjective micturition problems. Urological re-evaluation was performed two weeks and three months after nitrate medication. RESULTS: A significant improvement of peak urinary flow rates (+3.1 ml/s; p<0.05), IPS score and significant decrease of residual urine volume (-22 ml; p<0.05) were found in the symptomatic patients. No significant changes of micturition parameters were found in asymptomatic patients. PSA levels and prostate volumes did not change in either groups. CONCLUSIONS: Organic nitrates influence micturition parameters in patients with obstructive benign prostatic hyperplasia. This might be explained by the known mechanism of NO donation (smooth muscle relaxation) of nitrates. More functional controlled studies are necessary to describe the grade of influence of nitrates on the prostate. Concomitant oral medication with nitrates must be considered as a relevant bias factor on BPH in future clinical studies. PMID- 10672954 TI - Relationship between HLA-DR antigen and HLA-DRB1 alleles and prostate cancer in Japanese men. AB - PURPOSE: Human leukocyte antigens (HLA) are cell surface glycoproteins playing a key role in the immune system. In some cancers, changes in major histocompatibility complex (MHC) class I and II expression, usually a reduction or loss of these molecules, appear to provide a mechanism whereby tumour cells may escape host immunity. We investigated the relationship between HLA, especially class II, molecules and prostate cancer in Japanese men using molecular techniques. MATERIALS AND METHODS: HLA class II typing was performed by the polymerase chain reaction-sequence specific primer (PCR-SSP) method of analysis and/or a commercial rapid assay based on the PCR followed by reverse dot blot hybridization of the PCR products (Inno-LiPA assay). Allele frequencies were calculated. HLA allele frequencies reported in 1216 healthy Japanese individuals were used as the control data. Differences in allele frequency between subjects and the control group were analyzed by the chi-square test. The relationship between HLA antigens/alleles and prostate cancer is expressed in terms of relative risk (RR). RESULTS: The frequencies of HLA-DR4 were significantly higher in Japanese men with prostate cancer than in the healthy control group (gene frequency 36.2% vs. 26.3% in control, p<0.05), although the relative risk of prostate cancer was less than 2. Furthermore, the frequencies of HLA-DRB1-0406, 0410 and 1405 allele were significantly higher in the prostate cancer group than in the control group (allele frequency was 7.3%, 4.5% and 5.4% vs. 3.03%, 1.79% and 2.22%, p<0.05, respectively). RR of those HLA-DRB1 allele for prostate cancer was 2.6 in each allele. CONCLUSIONS: HLA molecules may be useful for the early detection of prostate cancer as a risk factor, and also for recognizing cancer activity by using them as a marker helpful in the choice of appropriate treatment by predicting prognosis. PMID- 10672955 TI - Improved discrimination of prostate cancer and benign prostatic hyperplasia by means of the quotient of free and total PSA. AB - The value of t-PSA (total prostate specific antigen) and of the quotient of free and t-PSA (% f-PSA) for the discrimination of BPH (benign prostatic hyperplasia) and PC (prostate cancer) as well as possible influencing factors were subject to examination under study conditions. The sera of 210 patients (131 BPH, 79 PC patients) were examined by means of the Immulite test; thereof 76 male patients (47 BPH, 29 PC patients) were found to have a t-PSA-value between 4 and 10 ng/ml (grey area). Apart from the age and the findings of rectal digital examination, we recorded the prostate volume, indications of non-specific increases in PSA and for PC patients also the TNM-G stage. For patients with prostate cancer the quotient of f- and t-PSA was significantly lower (median: 0.08) than compared to patients with BPH (median: 0.22) (p<0.001). Also in the grey area the quotient was significantly lower in patients with malignant diagnosis (median: 0.12) than for patients with a non-malignant diagnosis (median: 0.21) (p<0.001). ROC curves were prepared in order to compare the capability of discrimination of the two parameters. At this point, the better discrimination potential of the quotient in the grey area became evident. Due to the fact that priority was given to the detection of carcinoma, the threshold value was defined at a level at which high sensitivity (90%) is existent in combination with an acceptable specificity (approx. 50%). The resultant values are for the total PSA area 0.21, for the grey area 0.19 as a cut-off. Neither the age, nor the prostate volume, nor urinary tract infections had any influence upon the quotient. There was also no correlation between the stage or the grading of the tumour and the percentage of the f-PSA. The quotient alleviates the discrimination between BPH and PC, in particular in the diagnostically problematic grey area. Thus, it can serve as an aid for the decision "biopsy or re-biopsy". As there is currently no standardized method for the application of % f-PSA, there is a requirement for further examination under homogeneous criteria. PMID- 10672956 TI - Demonstration of testicular apoptosis in human male infertility states using a DNA laddering technique. AB - DNA laddering, morphometric analysis, and in-situ end labelling of testis biopsy tissue obtained from azoospermic or severely oligozoospermic men revealed increased apoptosis frequency, implicating a possible role of apoptosis in the pathogenesis of human male infertility. PMID- 10672957 TI - Echo-colour doppler ultrasonography in the diagnosis of varicocele. AB - The relationship between varicocele and infertility has long been defined. About a third of the male patients undergoing evaluation for infertility present with a varicocele. Sixty male patients between 17 and 35 years of age (mean 25.6) were examined with a colour-doppler flow imaging system. The diameters of the veins in the pampiniform plexus were measured by gray-scale sonography. Our findings were classified with regard to venous diameter, the existence or non-existence of reflux, the circumstances under which these findings were recorded (e.g. during normal respiration and standing position or during Valsalva manoeuvre and supine position). Finally our results suggest that: (a) the clinical significance of the presence of dilated veins or reflux during increased intraabdominal pressure and under similar circumstances should be regarded with caution; (b) positive findings during normal inspirium are highly significant (grades III and IV). PMID- 10672958 TI - Factors involved in diagnostic delay of testicular cancer. AB - In testicular cancer, which is one of the highly curable cancers, the beneficial effect of early diagnosis on survival has been well accepted for years. This study was planned to determine the probable factors involved in diagnostic delay of testicular cancer. A total of 140 patients with testicular cancer were included in this study. We attempted to find a relationship between the mode of presentation, stage of the tumour, patients' socioeconomic status and the delay in diagnosis. Majority of patients presented with scrotal pain (49.3%). No significant correlation was found between the mode of presentation and stage of the disease (p>0.05). The median time of delay in diagnosis was 5.4 months. The stage at presentation was not influenced by the delay in diagnosis. Similarly the annual income and the educational level of the patients had no significant impact on the delay in diagnosis. No effect of delay in diagnosis on the stage at presentation could be demonstrated in our study. Our results indicate that in this country people are not thoroughly educated about the importance of routine self-examination of the testes. It must also be emphasized that painful testicular masses need to be evaluated carefully to rule out malignancy. PMID- 10672959 TI - Immature germ cells in semen and their correlations with other semen parameters. AB - PURPOSE: The significance of the presence of leukocytes and immature germ cells in semen and other parameters of semen is currently a subject of controversy. MATERIALS AND METHODS: Semen from 572 subfertile patients was analyzed according to WHO criteria and leukocytes as well as immature germ cells were assessed by identifying the round cells in semen by peroxidase staining. Microbiological investigation was carried out in cases with leukocyte counts of >1,000,000/ml. RESULTS: It was found that as the concentration of spermatozoa decreased the rate of immature germ cells increased and this increase was accompanied by a decrease in motility and in the number of spermatozoa with normal morphology. As the sperm count increased, motility, number of spermatozoa with normal morphology and of immature germ cells also increased whereas an increase in sperm motility was accompanied by an increase in the percentage of spermatozoa with normal morphology. Microbiological investigations were negative in patients with semen leukocyte counts of >l,000,000/ml. CONCLUSIONS: Although it is possible to establish that the leukocyte and immature germ cell counts correlate with other parameters of semen, these correlations are not statistically significant. The most significant finding is that as the number of sperms decreases, the ratio of immature germ cells to total germ cells increases. While assessing immature germ cells instead of giving special attention to the number of immature germ cells in semen, the ratio of immature germ cells to total germ cells should be considered. The increase of leukocyte count in the semen of oligospermic patients may not always mean leukospermia. PMID- 10672960 TI - The significance of testicular reactive oxygen species on testicular histology in infertile patients. AB - This study was designed to investigate the relationship between the effects of testicular reactive oxygen species (ROS) levels and testicular histology on infertile patients with the aid of xanthine oxidase system and testicular tissue malondialdehyde levels. Forty patients with idiopathic infertility constituted our study group. Bilateral testicular biopsies were performed and spermatogenesis was assessed histopathologically. Patients were divided into 4 groups according to spermatogenic pattern (normal spermatogenesis; hypospermatogenesis; maturation arrest; Sertoli cell only syndrome). Testicular tissue xanthine oxidase and malondialdehyde (MDA) concentrations were analyzed in each sample by spectrophotometric assay and thiobarbituric acid reaction assay, respectively. Testicular tissue MDA and xanthine oxidase concentrations were not statistically different in patients having normal spermatogenesis, with respect to Sertoli cell only syndrome, maturation arrest and hypospermatogenesis, respectively. As a result of our study we think that there are still some factors other than ROS which may be important contributors to spermatogenetic injury that need to be examined. PMID- 10672961 TI - Relationship between zinc concentrations in seminal plasma and various sperm parameters. AB - The zinc concentration in seminal plasma from 98 infertile male patients and 8 fertile males was measured. The zinc concentration of the seminal plasma in azoospermic and oligoasthenozoospermic patients was significantly lower than that in the other groups (each, p<0.05 ). The seminal plasma zinc concentration in asthenozoospermic males was significantly higher than that in any other group (p<0.05). There was a positive correlation of zinc concentration with sperm concentration (r=0.33, p<0.05) and with sperm motility (r=0.22, p<0.05), while there was no correlation with sperm morphology. A correlation between zinc concentration and plasma testosterone concentration was observed (r=0.24, p<0.05). It is concluded that excessively high zinc concentration is apparently related to defective motility in asthenozoospermic patients, even though adequate seminal plasma content of the element is required for normal sperm function. PMID- 10672962 TI - Diagnostic application of AAP isoenzyme separation. AB - We describe the separation of alanylaminopeptidase (AAP) from urine into two isoenzymes: a particulate and a soluble form. The separation was accomplished using ion-exchange column chromatography on DEAE-52 cellulose. The purity of the isolated forms was confirmed electrophoretically. We attempted to create a method allowing the quantitative assessment of AAP isoenzymes in urine based on electrophoretic separation in 7.5% polyacrylamide gel with subsequent densitometric analysis. The content of AAP isoenzymes in examined urine was estimated using ultracentrifugation. The differences in the content of cytosolic and microsomal forms were observed suggesting the possibilities of using AAP isoenzymes in diagnostics. Furthermore, the effect of temperature on the activity of AAP separated on DEAE-52 cellulose was studied. PMID- 10672963 TI - The effects of dexamethasone on the response of bronchus-associated lymphoid tissue to intranasal administration of formalin-killed Pasteurella haemolytica A2 in goats. AB - A trial was conducted to observe the immediate and chronic effects in goats of dexamethasone administration on the bronchus-associated lymphoid tissue (BALT) response to intranasal administration of formalin-killed Pasteurella haemolytica A2. Twenty-four goats were divided into four groups. Those in group 1 were injected intramuscularly with 1 mg/kg dexamethasone on three consecutive days, followed by intranasal exposure to formalin-killed P. haemolytica A2 one day after the last dexamethasone treatment. The goats in group 2 were similarly injected with dexamethasone followed by intranasal exposure to formalin-killed P. haemolytica A2 21 days after the last dexamethasone treatment. The animals in group 3 were exposed intranasally to formalin-killed P. haemolytica A2 without prior dexamethasone treatment. The animals in group 4 were untreated controls. The intranasal exposures to formalin-killed P. haemolytica A2 were repeated 2 weeks later. Intranasal exposure to formalin-killed P. haemolytica 1 day after dexamethasone treatment further reduced the number and size of BALT compared to the untreated control. Significantly (p < 0.01) more reduction of BALT occurred in goats exposed to formalin-killed P. haemolytica A2 21 days after dexamethasone treatment. On the other hand, intranasal exposure of goats without prior dexamethasone treatment stimulated the BALT compared to the untreated controls. PMID- 10672964 TI - Preliminary studies on hepatic carnitine palmitoyltransferase in dairy cattle with or without fatty liver. AB - The hepatic mitochondrial carnitine palmitoyltransferase (CPT) activity was measured by fluorimetric assay in dairy cows with or without fatty liver. CPT activities in 13 lactating cattle and in 6 non-lactating cows were 304.4+/-86.6 micromol CoA/min per g protein and 169.3+/-84.8 micromol CoA/min per g protein, respectively. This difference was significant (p < 0.05). CPT activities in early lactation (0-110 days after calving), mid-lactation (111-220 days after calving) and late lactation (over 220 days after calving) were 278.9+/-68.0, 312.4+/-124.1 and 320+/-59.3 micromol CoA/min per g protein, respectively. There was no significant difference between the values at different stages of lactation. The CPT activity in 10 lactating cows with fatty liver unrelated to calving was 201.3+/-80.0 micromol CoA/min per g protein. CPT activity in 10 cattle with fatty liver was significantly lower than that in normal lactating cattle. Based on these findings, clinical fatty liver unrelated to calving appears to be associated with a decrease in hepatic CPT activity. PMID- 10672965 TI - The influences of dietary selenium and vitamin E intakes on milk somatic cell counts and mastitis in cows. AB - Dietary supplements of selenium and vitamin E in greater amounts than are required for nutritional adequacy can have complementary functions in reducing somatic cell counts and both the severity and duration of clinical mastitis. Selenium inadequacy is geographically widespread and can frequently be a year round problem. In contrast, an adequate intake of fresh grass and quality grass silage or other green, leafy material should provide adequate vitamin E. Many observations indicate that in farm situations where there is good udder hygiene and where long-acting antibiotic treatment is given at drying off, significant correlations are found between the mean bulk milk somatic cell counts and the blood selenium concentration or glutathione peroxidase activity in the blood, even where plasma vitamin E concentration is fully adequate. The accompanying reduced incidence of clinically affected quarters diminishes the need for corrective antibiotic treatment during lactation. Presentation of selenium and vitamin E within a sustained-release rumen bolus system during the dry period and into the succeeding lactation is a convenient means of supplementation to avoid over- or under-consumption by individual cows within a group. Adequate hygiene of the environment, the milking equipment and the udder are essential. PMID- 10672966 TI - Pharmacokinetics and dosage regimen of enrofloxacin in buffalo bulls after intramuscular administration. AB - The disposition kinetics and dosage regimen of enrofloxacin were investigated in breeding buffalo bulls following a single intramuscular administration of 5 mg/kg. The absorption half-life, half-life of the terminal phase, apparent volume of distribution and total body clearance were 0.262+0.099 h, 1.97+/-0.23 h, 0.61+/-0.13 L/kg and 210.2+/-18.6 ml/(kg.h), respectively. Therapeutic plasma levels (> or = 1 microg/ml) were maintained for up to 6 h. A satisfactory intramuscular dosage regimen for enrofloxacin in buffalo bulls would be 8.5 mg/kg followed by 8.0 mg/kg at 8 h intervals. PMID- 10672967 TI - Pharmacokinetics of amoxicillin trihydrate in Desert sheep and Nubian goats. AB - The pharmacokinetics of amoxicillin were studied in five Desert sheep and five Nubian goats after intravenous (i.v.) or intramuscular (i.m.) administration of a single dose of 10 mg/kg body weight. Following i.v. injection, the plasma concentration-versus-time data were best described by a two-compartment open model. The kinetic variables were similar in both species except for the volume of the central compartment (Vc), which was larger in sheep (p<0.05). Following i.m. injection, except for the longer half-life time of absorption in goats (p<0.05), there were no significant differences in other pharmacokinetic parameters between sheep and goats. The route of amoxicillin administration had no significant effect on the terminal elimination half-life in either species. The bioavailability of the drug (F) after i.m. administration was high (> 0.90) in both species. These results indicate that the pharmacokinetics of amoxicillin did not differ between sheep and goats; furthermore, because of the high availability and short half-life of absorption, the i.m. route gives similar results to the i.v. route. Therefore, identical intramuscular and intravenous dose regimens should be applicable to both species. PMID- 10672968 TI - A labelled avidin-biotin ELISA to detect antibodies to caprine arthritis encephalitis virus in goats' sera. AB - A labelled avidin-biotin ELISA (lab-ELISA) was developed and compared with indirect ELISA (i-ELISA) and agar-gel immunodiffusion assay (AGID) for its efficacy in detecting antibodies against caprine arthritis-encephalitis virus (CAEV) in goat sera. The enzyme immunoassays were standardized using 113 sera from CAEV-negative goat flocks. The tests were compared using the results from 339 serum samples. The lab-ELISA showed the greatest number of positive results (94/339) as compared with AGID (51) and i-ELISA (64). The comparison of the other two tests with the lab-ELISA showed an agreement of 87.3% with AGID and 90.6% with i-ELISA. The lab-ELISA may be useful for screening large populations for CAEV antibodies, in epidemiological surveys and in the control of caprine arthritis-encephalitis. PMID- 10672969 TI - Anxiolytic-like effects of 7-nitroindazole in the rat plus-maze test. AB - It is considered that nitric oxide (NO) is one of the most interesting research subjects. Because the actual role of NO in the mechanism of anxiety is still unclear, in this study, the involvement of NO in the mechanism of anxiety was investigated, using the plus-maze test. 7-Nitroindazole (7-NI) (15, 30, 60, 90, and 120 mg/kg), a new nitric oxide synthase (NOS) inhibitor was studied. The time spent on open arms and open-arm visits was evaluated. 7-NI, at 15-120 mg/kg doses potently increased the time spent on open arms and open-arm visits. However, at 120 mg/kg it attenuated the time spent on the open arms, compared to at 90 mg/kg. This effect was attributed to decreased locomotor activity in the higher dose group. Neither L-arginine, nor D-arginine (100 mg/kg) significantly affected any of the behavioral parameters measured in the rat elevated plus-maze test. Neither drugs revealed any effect on locomotion. L-Arginine but not D-arginine given 10 min before 7-NI, reversed the 7-NI induced anxiolytic-like effects. These data support an involvement of NO in the process of anxiety, and further suggest that the anxiolytic-like effect of 7-NI may be attributable to the inhibition of NO synthesis. PMID- 10672970 TI - Turning in rats following intraaccumbens shell injections of amphetamine or eticlopride. AB - In recent years there has been interest in possible differential behavioral functions of the core and shell subregions of the nucleus accumbens. The present study compared the effects of accumbens core and shell injections of (+) amphetamine on turning behavior in rats; the turning effects of the selective D2 antagonist (-)-eticlopride injected into the shell were also tested. Rats (n = 28) were implanted unilaterally with guide cannulae and subsequently tested for turning in seven 20-min sessions. Amphetamine (10.0 and 20.0 microg but not 5.0 microg/ 0.5 microl) elicited contralateral turning following injection into the shell, while the same doses into the core had no effect. In animals given systemic amphetamine (2.0 mg/kg), eticlopride (10.0 but not 1.0 or 0.1 microg/0.5 microl) injected into the shell region of the accumbens produced ipsilateral turning. These results suggest that the accumbens shell, but not the core, is a critical site for turning behavior. PMID- 10672971 TI - Acute and chronic effects of gepirone and fluoxetine in rats tested in the elevated plus-maze: an ethological analysis. AB - The potential role of 5-hydroxytryptamine (5-HT) in anxiety has been the subject of much research, most of it addressed to the hypothesis that 5-HT promotes anxiety and, therefore, that drugs that reduce 5-HT functions will be effective anxiolytic agents in human anxiety disorders. However, the effects of serotoninergic drugs in different behavioral paradigms have been inconsistent. These inconsistencies have been particularly well illustrated in the elevated plus-maze. In the present study we provided an ethopharmacological analysis (in addition to conventional measures) of the behavior of rats in the elevated plus maze with transparent walls after acute and chronic treatments with gepirone, an agonist of 5-HT1A receptors, and fluoxetine, a selective inhibitor of serotonin reuptake. Although gepirone has been used to treat anxiety, fluoxetine is a mainstay in the treatment of depression. Acute treatment with gepirone (1, 3, 5.6, and 10 mg/kg, IP) produced an anxiogenic profile with increased risk assessment behaviors (e.g., flat-back approach) and decreased behavioral measures that are inversely related to "anxiety" (e.g., head dipping and end-arm activity). In contrast, chronic gepirone (10 mg/kg day, PO) produced an opposite effect showing an anxiolytic profile that is consistent with the clinical use of this drug, which shows efficacy after 2-4 weeks of treatment. Acute fluoxetine (5.6 and 10 mg/kg, IP) also produced an anxiogenic profile with reduced head dipping and end-arm activity. On the other hand, chronic fluoxetine (10 mg/kg day, PO) had no effect on any of the behavioral measures. These data demonstrate: (a) the anxiogenic and anxiolytic effects of acute and chronic gepirone, respectively, corroborate with the observed effects of these treatments in the clinic; (b) similarly, the anxiogenic effects of acute fluoxetine observed here have also been reported in clinical studies with 5-HT reuptake blockers. This class of compounds has not been systematically used as anxiolytic; (c) the elevated plus-maze with transparent walls shows good sensitivity for evaluating serotonergic drugs with anxiogenic and anxiolytic profile. PMID- 10672972 TI - Role of brain dynorphin in nitrous oxide antinociception in mice. AB - Earlier studies indicate that nitrous oxide antinociception is mediated by opioid receptors, and we have hypothesized that nitrous oxide stimulates a neuronal release of an endogenous opioid peptide (EOP) that stimulates opioid receptors. To further test this hypothesis, male NIH Swiss mice were pretreated intracerebroventricularly with rabbit antisera to opioid peptides or with various inhibitors of peptidases involved in the degradation of EOPs. Mice were subsequently exposed to three different concentrations of nitrous oxide in oxygen, and their antinociceptive responsiveness was measured using the acetic acid abdominal constriction test. Nitrous oxide antinociception was significantly attenuated by 24-h pretreatment with antisera to various fragments of dynorphin (DYN) but not by antisera against methionine-enkephalin (ME) or beta-endorphin (beta-EP). In other experiments, nitrous oxide antinociception was significantly enhanced by 30-min pretreatment with phosphoramidon, an inhibitor of endopeptidase 24.11, which has been implicated in DYN degradation, but not bestatin or captopril, which inhibit aminopeptidase and angiotensin-converting enzyme, respectively. The latter enzymes have been implicated in degradation of certain EOPs albeit not DYN. These findings support the hypothesis that nitrous oxide antinociception in the mouse abdominal constriction test is mediated by endogenous DYN acting in the central nervous system. PMID- 10672973 TI - The role of the benzodiazepine-GABA system in the memory processes of the day-old chick. AB - This series of experiments investigated the effect of the benzodiazepine diazepam on memory formation in day-old chicks trained on a single-trial, passive avoidance task. The findings indicate that diazepam has a dose-specific and time dependent effect on memory processes. A 0.125-mg/kg dose of diazepam administered immediately after training led to amnesia in these subjects only after 30 min following learning. Pretreatment with bicuculline and flumazenil were effective in ameliorating the memory deficits caused by diazepam, and consolidated memory function in saline-treated controls following strong and weak aversant training. These findings suggest that benzodiazepine effects on memory are mediated by their effects on arousal, possibly by the release of noradrenaline, which is critical to the establishment of long-term memory. PMID- 10672974 TI - Serotonergic deficits and impaired passive-avoidance learning in rats by MDEA: a comparison with MDMA. AB - The serotonergic deficits induced by 3,4-methylenedioxyethamphetamine (MDEA, "eve"), were examined and compared with 3,4 methylenedioxymethamphetamine (MDMA, "ecstasy"). A single dose of MDEA (10, 20, or 40 mg/kg IP) induced a dose-related hyperthermia, but only the highest dose significantly reduced 5-HT content and 5 HT transporter density in the frontal cortex and in the hippocampus 7 days later. Long-term serotonergic deficits were much more marked when MDEA was given repeatedly (40 mg/kg IP., b.i.d., for 4 consecutive days). Single or repeated administration of MDEA induced no change on 5-HT1A receptor density in the frontal cortex, brain stem, or hippocampus, although 3 h after both treatments plasma corticosterone levels were significantly increased. MDEA (5-20 mg/kg, IP) produced significant retention deficits in a passive-avoidance learning task. Conversely, 7 days after the repeated administration of MDEA (40 mg/kg b.i.d., for 4 consecutive days) no effect on passive-avoidance performance was observed unless rats were treated again with another dose of MDEA (20 mg/kg IP) 30 min before the training trial. The 5-HT1A receptor antagonist, WAY 100635, prevented the impairment in retention performance induced by 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), but not by MDEA or MDMA, indicating that the effect of these amphetamine derivates was not mediated by 5-HT1A receptor activation. The results suggest the risk of serotonergic dysfunction associated with MDEA abuse in humans. PMID- 10672975 TI - Repeated treatments with 7-OH-DPAT: context-independent behavioral sensitization and conditioned hyperactivity. AB - The primary objective of this study was to determine whether the expression of behavioral sensitization to the putative dopamine D3 receptor agonist 7-OH-DPAT is context dependent. Three groups (n = 8 each) of male Wistar rats (250-350 g) were given nine injections (at 48-h intervals) of 7-OH-DPAT (1.0 mg/kg, SC) or vehicle 15 min before and after activity testing. The paired group received 7-OH DPAT before activity testing and vehicle after testing. The unpaired group received vehicle before and 7-OH-DPAT after testing, and the vehicle control group received two vehicle injections. Locomotor activity was measured in photocell arenas for 2 h. After the first seven sessions, all rats were tested for activity following a vehicle injection to test for possible conditioning effects. Prior to the 11th session, all rats were given a challenge injection of 7-OH-DPAT (1.0 mg/kg, SC) to test for sensitization. Major findings were as follows: (a) the 7-OH-DPAT/paired group displayed a progressively greater increase in locomotor activity with repeated treatments; (b) the 7-OH-DPAT/paired group was significantly more active than either the vehicle control group or the 7-OH-DPAT/unpaired group during the vehicle test session; and (c) after the 7-OH DPAT challenge injection, the paired and unpaired 7-OH-DPAT groups were significantly, and equally, more active than the vehicle control group. In contrast to previous findings with the D2-type dopamine agonists bromocriptine and quinpirole, these results suggest that the expression of behavioral sensitization to 7-OH-DPAT is not context dependent. Moreover, these results suggest that the apparent conditioned hyperactivity and context dependency often observed after repeated dopamine agonist treatments may not be related to the same associative and/or nonassociative mechanisms. PMID- 10672976 TI - Effects of acute nicotine on several operant behaviors in rats. AB - The present experiment assessed nicotine's effects on complex cognitive processes using a variety of operant tasks in rats, including incremental repeated acquisition (IRA) to assess learning; conditioned position responding (CPR) to assess auditory, visual, and position discrimination; progressive ratio (PR) to assess motivation; temporal response differentiation (TRD) to assess timing; and differential reinforcement of low response rates (DRL) to assess timing and response inhibition. Acute nicotine administration (0.0, 0.3, 0.42, 0.56, 0.75, and 1.0 mg/kg, IP) increased IRA and CPR response rate without significantly altering accuracy. Nicotine had similar effects on response rate for PR. For TRD, nicotine had a U-shaped dose effect on accuracy, but failed to shift the mode of the TRD response distribution. For DRL, nicotine reduced accuracy and also shifted the mode of the DRL response initiation time distribution to the left. Nicotine produced an inverted U-shaped dose-effect curve for the overall number of "bursting" responses under both of these schedules. The results of this experiment suggest that nicotine can impair performance on some aspects of cognitive-behavioral performance, while simultaneously improving performance on others. PMID- 10672977 TI - Effects of labetalol treatment on the physiological and subjective response to smoked cocaine. AB - Adrenergic receptors mediate some of the physiological and possibly behavioral effects of cocaine. The purpose of this study was to investigate the effect of treatment with a peripherally acting adrenergic blocking drug labetalol on the cardiovascular and subjective response to repeated deliveries of smoked cocaine. In this double-blind, placebo-controlled, crossover study, 12 cocaine users were treated with a single 100 or 200 mg dose of labetalol, or placebo in each of three experimental sessions. Starting 2 h after the medication treatment, subjects received three doses of 0.4 mg/kg smoked cocaine, 30 min apart. Labetalol treatment significantly attenuated the cocaine-induced increases in heart rate and systolic blood pressure. This effect of labetalol on the cardiovascular response did not decrease with repeated cocaine deliveries. The subjective response to smoked cocaine deliveries was not affected by labetalol treatment. These results suggest that labetalol effectively attenuates the systolic blood pressure and heart rate increases induced by repeated doses of smoked cocaine, but does not alter subjective effects. PMID- 10672978 TI - Changes in analgesia-producing mechanism of repeated cold stress loading in mice. AB - Functional changes in opioid receptors involved in analgesia of repeated cold stress (RCS)-loaded mice were investigated. The antinociceptive potency of morphine (4 mg/kg, PO) was not affected in normal mice by norbinaltorphimine (10 mg/kg, SC), but treatment with this agent resulted in a lower level of morphine induced antinociception in RCS-loaded animals. The antinociceptive activity of U 50488H (3 mg/kg, SC) was increased in RCS-loaded mice. In contrast to hypersensitivity to U-50488H (1 and 10 microg, IT) noted in RCS-loaded mice, the antinociception induced by DAMGO (0.1 and 1 microg, ICV) was reduced compared to that of normal animals. Diazepam (1 mg/kg/day SC) was given during RCS loading, and this agent prevented the development of hyperalgesia and the decrease in the antinociceptive activity of DAMGO (1 microg, ICV) in RCS-loaded mice, but there was no effect on the enhancement of the antinociceptive potency of U-50488H (10 microg, IT). These results indicate that the RCS-loaded mice were hyposensitive to supraspinal mu-opioid receptor-mediated antinociception, whereas their antinociceptive activities through kappa-opioid receptor in the spinal cord were increased. Hypofunction of the supraspinal mu-opioid receptor due to anxiety may explain the mechanism involved in the lowering of the nociceptive threshold in RCS-loaded animals. PMID- 10672979 TI - Pontine tegmentum lesions increase anxiety-like behavior in rats: a comparison with anxiety produced by beta-CCE. AB - Electrolytic lesions of the pedunculopontine tegmental nucleus (PPTg) have been previously reported to increase anxiety-like behavior in rats. The aim of the present study was to compare these behavioral changes with those produced by an anxiogenic compound, the partial inverse agonist at benzodiazepine receptors, beta-CCE. Three groups of rats, sham-lesioned treated with vehicle, sham-lesioned treated with 10 mg/kg of beta-CCE, and PPTg-lesioned rats treated with vehicle, were tested in the elevated plus-maze, the social-interaction test, and for spontaneous locomotion. Histology showed that lesions were concentrated on the caudal half of the PPTg. Measures of both the PPTg-lesioned and beta-CCE-treated rats indicated increased anxiety-like behavior in the elevated plus-maze and in the social-interaction test. Spontaneous locomotion, measured in the open- field arena, did not differ between sham controls and PPTg-lesioned rats, but was decreased in rats treated with beta-CCE. Our results confirmed that electrolytic lesions of the caudal PPTg produce increased anxiety-like behavior. This behavior is quantitatively and qualitatively similar to that produced by 10 mg/kg of beta CCE. PMID- 10672980 TI - Effects of adrenoceptor agents on apomorphine-induced licking behavior in rats. AB - In the present study, intraperitoneal (IP) administration of the dopaminergic receptor agonist apomorphine (0.1, 0.25, and 0.5 mg/kg) induced a dose-dependent licking in rats. The intraperitoneal injection of the alpha1'''adrenoceptor agonist phenylephrine (1-8 mg/kg) but not the alpha2-adrenoceptor agonist clonidine (0.025-0.05 mg/kg) decreased licking induced by apomorphine. The alpha adrenoceptor antagonists prazosin, phenoxybenzamine, and yohimbine also reduced the apomorphine response significantly. The response induced by phenylephrine was decreased by a dose of prazosin. The beta1-adrenenocepor agonist dobutamine and beta2-adrenenocepor agonist salbutamol did not alter the apomorphine response. However, beta2-adrenenocepor antagonists atenolol and propranolol reduced the apomorphine effect. It may be concluded that alpha1- and possibly beta1 adrenoceptor mechanisms may be involved in modulation of licking behavior. PMID- 10672981 TI - Ethanol, but not the anxiolytic drugs buspirone and diazepam, produces a conditioned place preference in rats exposed to conditioned fear stress. AB - The present study was designed to investigate the role of an anxiolytic effect in the development of a drug-associated place preference in rats exposed to conditioned fear stress, using the conditioned place-preference paradigm. The administration of a low dose of ethanol (300 mg/kg, IP) and the anxiolytic drugs, buspirone (1 and 2 mg/kg, IP) and diazepam (1.25 and 2.5 mg/kg, IP), did not produce a place preference in rats that were not exposed to conditioned fear stress. In rats that were exposed to conditioned fear stress, ethanol produced a significant place preference, while buspirone and diazepam failed to produce a place preference. In addition, ethanol, buspirone, and diazepam produced no place preference in rats treated with an anxiogenic dose of pentylenetetrazole (20 mg/kg, IP). A significant decrease in locomotor activity was observed in rats exposed to conditioned fear stress. Ethanol, but not buspirone and diazepam, significantly recovered or increased locomotor activity in rats exposed to conditioned fear stress. Further, the locomotor-stimulating effect of ethanol was markedly enhanced by repeated exposure to conditioned fear stress. These results suggest that the stimulating effect may be strongly related to the development of the rewarding effect of a low dose of ethanol under psychological stress, and that the conditioned place preference paradigm with conditioned fear stress may be useful for studying the rewarding mechanism of ethanol with regard to the interaction between ethanol and psychological stress. PMID- 10672982 TI - Characterization of cocaine self-administration and pharmacokinetics as a function of time of day in the rat. AB - Two experiments examined the influence of time of day on the intravenous self administration of cocaine and its associated pharmacokinetic profile in male Sprague-Dawley rats. In both experiments, individual rats were randomly assigned to experimental groups (n = 6/group) according to four selected times of day, 0100, 0700, 1300, and 1900 h, during which experimental procedures were conducted. In both experiments, rats were maintained under a 12 L:12 D ambient lighting cycle, with lights on at 0600 h. Training and testing was thus conducted either 1 (0700, 1900) or 7 (1300, 0100 h) hours into the light and dark phases. In Experiment 1, characteristics of cocaine self-administration across a behaviorally active dose range were assessed. Statistically significant differences were observed in the rates and patterns of self-administration across the four experimental groups, most notably characterized by an apparent shift in the dose of cocaine, which engendered peak rates of responding. Specifically, groups tested at 0100 and 1300 h appeared to exhibit enhanced sensitivity to the reinforcing properties of low-dose cocaine relative to groups tested at 0700 and 1900 h. The observed differences in apparent sensitivity of experimental subjects to low-dose cocaine were not related in any simple way to ongoing patterns of general locomotor activity, and were not accompanied by corresponding variance in the pharmacokinetic profiles of cocaine when assessed over 1 h following an intravenous infusion (1.8 mg/kg) at each of the four sampling periods noted above. PMID- 10672983 TI - Fear-potentiated startle response in mice: genetic analysis of the C57BL/6J and DBA/2J intercross. AB - The role of genetic factors in the fear-potentiated startle (FPS) response was examined in the inbred C57BL/6J (B6) and DBA/2J (D2) mouse strains. Mice in the D2 strain displayed a significant potentiation in the acoustic startle response (ASR) when presented with a visual condition stimulus (CS) previously paired with an aversive unconditioned stimulus (US). The maximal FPS response was observed following 20 conditioning trials but a near maximal response was noted following as few as five trials. Forty conditioning trials produced a significant reduction in the FPS response that may be related to overtraining. The FPS response in the B6 strain was significantly lower than the D2 strain, regardless of the number of conditioning trials. The contrasting FPS responses were not related to differences in auditory sensitivity known to exist between these strains. Analysis of a full Mendelian cross formed from the B6 and D2 strains found that the FPS response was a highly heritable trait, best described by a simple additive model of inheritance and with a broad-sense heritability of 0.46. The distribution of the FPS response in F2 hybrids formed from the intercross of the D2 and B6 strains was continuous which suggests a multigenic substrate. The light + noise and noise-alone trial types were highly correlated, but no association was detected between the baseline ASR amplitude and the FPS response. Mice from the phenotypic extremes of the F2 distribution displayed FPS responses that were more extreme than either of the progenitor strains. However, both baseline startle amplitude and the salience of auditory stimuli did not differ in these groups. The results of this study confirm an early report by Falls et al. (1997), and provide additional quantitative genetics information necessary for the eventual mapping of the chromosomal regions or genes associated with the FPS response in mice. PMID- 10672984 TI - 6-Chloro-3'-nitroflavone is a potent ligand for the benzodiazepine binding site of the GABA(A) receptor devoid of intrinsic activity. AB - 6-Chloro-3'-nitroflavone integrates a list of nearly 70 flavone derivatives synthesized in our laboratories. The effects of 6-chloro-3'-nitroflavone on the benzodiazepine binding sites (BDZ-BSs) of the GABA(A) receptor were examined in vitro and in vivo. 6-Chloro-3'-nitroflavone inhibited the [3H]flunitrazepam ([3H]FNZ) binding to rat cerebral cortex membranes with a Ki of 6.68 nM and the addition of GABA to extensively washed membranes did not modify its affinity for the BDZ-BSs (GABA-shift = 1.16+/-0.12). The binding assays performed in rat striatal and cerebellar brain membranes showed that this compound has similar affinity to different populations of BDZ-BSs. Electrophysiological experiments revealed that 6-chloro-3'-nitroflavone did not affect GABA(A)-receptors (GABA(A) Rs) responses recorded in Xenopus oocytes expressing alpha1beta2gamma2s subunits, but blocked the potentiation exerted by diazepam (DZ) on GABA-activated chloride currents. In vivo experiments showed that 6-chloro-3'-nitroflavone did not possess anxiolytic, anticonvulsant, sedative, myorelaxant actions in mice or amnestic effects in rats; however, 6-chloro-3'-nitroflavone antagonized diazepam induced antianxiety action, anticonvulsion, short-term, and long-term amnesia and motor incoordination. These biochemical, electrophysiological, and pharmacological results suggest that 6-chloro-3'-nitroflavone behaves as an antagonist of the BDZ-BSs. PMID- 10672985 TI - Facilitative effects of EGb 761 on olfactory recognition in young and aged rats. AB - The aim of the present study was to evaluate the effects of chronic and acute treatment by the Gingko biloba extract, EGb 761 (IPSEN, France) on olfactory short-term memory in rats, using a spontaneous recognition procedure. The effects of a daily EGb 761 treatment (30 or 60 mg/kg) over a period of 30 days (Experiment 1) were evaluated in young male rats. Those of a single injection of EGb 761 were assessed either in young male rats at 60 or 120 mg/kg (Experiment 2) or in aged female rats at 60 mg/kg (Experiment 3). Results showed that, at the highest dose (60 mg/kg), chronic EGb 761 treatment enhanced the recognition performances, allowing recognition at delays at which control animals did not show any recognition. Acute treatment enhanced recognition at both doses tested. The results of the third experiment showed that EGb 761 had an overall enhancement effect on the performances of aged rats. In summary, our results provide evidence for a short-term memory enhancement effect of EGb 761 in both young and aged rats. PMID- 10672986 TI - Rewarding properties of testosterone in intact male mice: a pilot study. AB - The present study examined the rewarding properties of 4-androsten-17beta-ol-3 one testosterone in intact male mice using the conditioned place preference (CPP) technique. In Experiment 1, the pharmacokinetics of 0.8 and 1.2 mg/kg of testosterone were studied to determine the most appropriate temporal interval to test behavior. Additionally, the locomotor activity was recorded to control a possible interfering effect on CPP. The maximum testosterone concentration was registered at 45 min of administration, and no effects on activity were found. In Experiment 2, three groups of male OF-1 mice received four pairings of the least preferred compartment with testosterone (0.8, 1, or 1.2 mg/kg, SC) for 30 min. On alternate days the preferred compartment was paired with vehicle for 30 min. The control group received vehicle in both compartments. No significant differences between groups were found in the time spent in the drug-paired compartment. However, when separate analyses were performed in conjunction with the color of the drug-paired compartment. CPP was observed only in animals pairing testosterone/black compartment. These results suggest that rewarding properties of testosterone treatment can be observed in male mice; these effects probably being dependent on the environmental cues used as conditioned stimuli. PMID- 10672987 TI - Mechanisms of chloroquine-induced body-scratching behavior in rats: evidence of involvement of endogenous opioid peptides. AB - Chloroquine is commonly used in the chemotherapy of malaria fever, and as an antiinflammatory disease-modifying agent in patients with rheumatoid arthritis or systemic lupus erythematosus. Administration of chloroquine (20.0 mg/kg IP) significantly (p < 0.05) increased the frequency of body scratching in rats to 29.5+/-9 in 30 min, compared to saline control animals (6.5+/-2/30 min). Morphine, a mu-opiate receptor agonist (1.0 mg/kg IP), potentiated the chloroquine-induced rat body scratching to 40+/-6.6, while the mu-opiate receptor antagonist, naltrexone (0.25 mg/kg, IP, given 15 min prior) blocked the chloroquine induced body scratching to 4.5+/-2 (p < 0.05 ANOVA). In addition, the frequency of chloroquine (20.0 mg/kg IP)-induced body scratching was significantly reduced to 9.1+/-3 in 30 min in rats rendered tolerant to morphine (p < 0.05 ANOVA) compared to the scratching frequency of 40+/-6.6 in morphine naive rats. These suggests an involvement of mu-opioid receptors and/or endogenous opioid peptides in chloroquine induced body scratching in rats. Promethazine, a histamine-receptor antagonist (1.0 mg/kg IP, given 15 min prior to chloroquine) and the corticosteroid, dexamethasone (1.0 mg/kg, IP, given 15 min prior) separately and significantly (p < 0.01) inhibited the chloroquine induced scratching in rats, in a similar manner to clinical studies in malaria. Collectively, the novel results implicate opioidergic mechanisms, and confirm the efficacy of antihistamine and corticosteroids in chloroquine body scratching in rats. It also strongly suggests that the chloroquine-induced body-scratching behavior in the rat may be a useful experimental model for chloroquine-induced pruritus in humans. PMID- 10672988 TI - Anxiolytic-like effects of antidepressants after acute administration in a four plate test in mice. AB - The four-plate test (FPT) is an animal model of anxiety based on stress caused by unconditioned fear. An increase of spontaneous punished behavior was used as a measure to determine the anxiolytic effects of various antidepressants (ADs). In the present study. ADs with different mechanisms of action, including tricyclics, selective serotonin reuptake inhibitors (SSRIs), a monoamine oxidase inhibitor (MAOI), and atypicals, were studied in the FPT to evaluate their anxiolytic-like effects following acute administration. The number of punished crossings was dramatically increased by the SSRIs citalopram, fluvoxamine, and paroxetine, but not fluoxetine. The mixed 5-HT/NE reuptake inhibitors, milnacipran and venlafaxine, also demonstrated strong antipunishment effects. The specific NE reuptake inhibitors, desipramine and maprotiline, and the atypical AD trazodone, enhanced freezing behavior, suggesting anxiogenic-like behavior. It was concluded that, in the FPT, a model based on spontaneous response, where animals are exposed to an aversive environment from which they can only escape by being motionless, this kind of behavior might be related to anticipatory anxiety. In this situation, ADs acting preferentially on 5-HT transmission possessed clear anxiolytic like effects. The balance between the two transmitters, 5-HT and NE, seemed to be a crucial factor. PMID- 10672989 TI - Taurine blocks the glutamate increase in the nucleus accumbens microdialysate of ethanol-dependent rats. AB - During ethanol withdrawal, dramatic changes in the concentration of many neurotransmitters may be responsible for many of the adverse effects. In the present study, the technique of microdialysis was used to assay the changes in excitatory and inhibitory amino acids after withdrawal from chronic ethanol intoxication. Rats were made physically dependent on ethanol by vapor inhalation for 4 weeks. The basal concentrations of both arginine and GABA were significantly decreased in ethanol-dependent rats, although there were no significant changes in any of the other amino acid basal concentration assayed (i.e.. glutamate and taurine). During the first 12 h after withdrawal from ethanol, only glutamate increased significantly (p < 0.05) at 6 h, and for the duration of the study period of 12 h. To investigate whether either taurine and ethanol interact with amino acids during ethanol withdrawal, two other ethanol dependent groups were injected with a single intraperitoneal injection of either taurine or ethanol 5 h after commencement of ethanol withdrawal. The IP injection of ethanol (2 g/kg) significantly increased taurine microdialysate content, and although this dose of ethanol was not able to block completely the increase of glutamate release after ethanol withdrawal, a delayed decrease in glutamate content was observed by the end of the period of the study (i.e., 11-12 h). However, IP injection of taurine (45 mg/kg) significantly blocked the increased glutamate release during ethanol withdrawal. This latter finding suggests that taurine may interact with glutamate, possibly by inducing a blockade of glutamate release during ethanol withdrawal. PMID- 10672990 TI - Behavioral effects of GABA(A) receptor stimulation and GABA-transporter inhibition. AB - The present analysis addressed behavioral changes after treatment with 4.5 mg/kg or 18.5 mg/kg of the GABA-uptake inhibitor tiagabine combined with either the benzodiazepine diazepam (1.5 mg/kg) or the imidazopyridine zolpidem (0.05 mg/kg), the latter two acting differentially on GABA(A) receptor subtypes. The study included 97 male PVG/OIaHsd rats. A standard open field, an enriched open field, and an elevated plus-maze was used to study rat behavior. Treatment with the low dose of tiagabine alone induced no specific behavioral effects, whereas the high dose had an anxiolytic-like potential. Furthermore, diazepam but not zolpidem displayed anxiolytic-like effects. Combination of each benzodiazepine receptor agonist with tiagabine at the low dose decreased explorative activity. Diazepam plus the high dose of tiagabine increased the activity in the open-field test. Zolpidem together with 18.5 mg/kg tiagabine had an angiogenic-like effect compared to pure tiagabine treatment. These results provide evidence for a pharmacodynamic interaction between the GABA-uptake inhibitor tiagabine and diazepam or zolpidem. The interaction might be relevant in the clinic when combining the anticonvulsant tiagabine and a benzodiazepine receptor agonist. PMID- 10672991 TI - Long-lasting cytoprotection after pentadecapeptide BPC 157, ranitidine, sucralfate or cholestyramine application in reflux oesophagitis in rats. AB - Recently, the effectiveness of pentadecapeptide BPC 157 and other anti-ulcer agents, called 'direct cytoprotection', was evidenced in totally gastrectomized rats duodenum challenged with cysteamine 24 h after surgery, and sacrificed 24 h after ulcerogen application. The further focus was on the possibility that this effect could be seen over a more prolonged period (1, 2, 4 weeks), and in other parts of the gastrointestinal tract (i.e. oesophagus). After the removal of the stomach, the oesophagus and jejunum were joined by a termino-lateral anastomosis. The animals were euthanized 7, 14 or 28 d after surgery, when oesophagitis was blindly assessed both macroscopically (percentage of ulcerations areas) and microscopically (percentage of areas of ulcers, regeneration and hyperplasia; number of inflammatory cells - polymorphonuclear and mononuclear). Starting 24 h after surgery, the medication was continuously given in the drinking water, in a volume of 12.5 mL/rat daily, until euthanasia at the end of the observation period, i.e. 7, 14, 28 d following surgery. Based on previous experiments, the doses of agents were daily calculated per kg b.w. as follows: BPC 157 125 mg or 125 ng, cholestyramine 2.5 mg, ranitidine 125 mg, sucralfate 725 mg, whereas controls received 72.5 mL x kg(-1) water. In support of these initial findings, and considering gastrectomized acid-free rats as an ideal model for long-term cytoprotective studies as well, pentadecapeptide BPC 157 markedly attenuated termino-lateral oesophagojejunal anastomosis-reflux oesophagitis also over a quite prolonged period. This efficacy was only partly shared by other anti-ulcer agents. After 1-week-old oesophagitis (microscopical assessment), but not after 2 or 4 weeks, less damaged mucosa was noted in rats drinking ranitidine or sucralfate compared to controls. Similar effectiveness was noted for cholestyramine. The obtained results were supported also by inflammatory cell assessment. Compared with control values, BPC 157-treated groups consistently presented less polymorphonuclears and less mononuclears in all assessed periods. Interestingly, the values obtained in other treated groups showed no difference compared with control values. Thus, despite limitations, a generalization supporting a direct importance of a common cytoprotective approach, could be clearly provided. A useful, long-lasting cytoprotective activity (apparently more prominent in BPC 157 rats, than in reference agents, ranitidine, sucralfate, as well as cholestyramine) may be a likely suitable therapy in otherwise resistant reflux oesophagitis conditions. PMID- 10672992 TI - The effect of pentadecapeptide BPC 157, H2-blockers, omeprazole and sucralfate on new vessels and new granulation tissue formation. AB - A clear protection of the gastrointestinal tract and an evident anti-inflammatory effect were shown for a novel stomach pentadecapeptide BPC 157 (i.p./i.g.) in comparison with several reference standards in various ulcer models along with a protection of endothelium and particular interaction with the NO-system. Thus, we evaluated whether this pentadecapeptide along with other gastroprotective agents could affect angiogenesis and the healing process in vivo using a procedure initially described by Szabo and co-workers. In each rat, two sterile sponges (1 x 1 x 0.25 cm; V = 0.25 mL) with the same quantities of BPC 157 (10 ng x mL(-1), 10 microg x mL(-1), 50 microg x kg(-1)) or reference agents (cimetidine: 10, 100, 500 mg x mL(-1); ranitidine: 2.5, 25, 250 mg x mL(-1); famotidine: 10, 50, 100 mg x mL(-1); omeprazole: 10, 50, 100 mg x mL(-1); sucralfate: 1, 5, 10 mg x mL(-1) were implanted subcutaneously in the lumbar region. The sponges were removed after 3 or 7 d, fixed in formalin, and processed for histologic and histochemical evaluation and morphometry assessment. Compared with the control values, the number of newly formed endothelial spaces inside newly formed granulation tissue was markedly increased in all animals treated with BPC 157, cimetidine, ranitidine, famotidine, sucralfate and omeprazole, a consistent finding noted after either 3 or 7 d. Compared with control values, markedly more granulation tissue was noted in the rats in the groups of animals treated with BPC 157 (50 microg) and in the rats treated with sucralfate in all dosages used, euthanized after 3 d. In all groups treated with H2-blockers however, similar values to those of controls were noted. Thus, it could be concluded that an evident angiogenic property was consistently noted for the novel pentadecapeptide BPC 157, H2-blockers (cimetidine, famotidine and ranitidine) and omeprazole, besides the well known angiogenic effect of sucralfate. Furthermore, unlike H2-blockers and omeprazole, BPC 157 stimulates the formation of granulation tissue, suggesting a particular activity, similar to that previously noted for sucralfate. PMID- 10672993 TI - Difference between the effect of acute and chronic surgical vagotomy on the cytoprotective action of atropine against indomethacin-induced mucosal lesions on the gastrointestinal tract in rats. AB - The cytoprotective effect of a small dose of atropine was proved against the indomethacin (IND)-caused gastrointestinal (GI) mucosal damage. This protective effect of atropine disappeared in the acute phase of surgical vagotomy (ASV) on the vagally-innervated parts of GI tract. The aims of our observations were: 1) to examine the effect of chronic surgical vagotomy (CSV) on the cytoprotective action of atropine in the GI tract; and 2) to compare the effects of ASV and CSV on the GI cytoprotection caused by atropine against IND-induced mucosal damage and vascular permeability in rats. The IND was given s.c. 24 h prior to the killing of the animals in a dose of 20 mg x kg(-1). Bilateral surgical vagotomy or sham operation were carried out 24 h (ASV) or 14 d (CSV) before IND application. Atropine was given i.p. every 5 h after IND-treatment in a dose of 0.1 mg x kg(-1). The number of macroscopical mucosal ulcerations was noted and its severity was calculated by semiquantitative scale in the stomach, small intestine and three equal parts of colon. Vascular permeability was measured by Evans-blue leakage into the mucosal tissue. It has been found that: 1) Tte small dose of atropine significantly decreased the IND-induced mucosal damage and vascular permeability on the stomach, small intestine and the vascular permeability on the proximal colon; 2) the small dose of atropine did not cause any changes in the appearance of IND-induced mucosal lesions and in Evans blue concentration in the mucosa after ASV, but it significantly decreased the IND caused mucosal damage and Evans blue concentration in the mucosa of stomach, small intestine and proximal colon after CSV; 3) the IND-induced mucosal damage and vascular permeability treated with atropine (given in cytoprotective dose) were significantly smaller after CSV than that after ASV on the stomach, small intestine and proximal colon. It has been concluded that the intact vagal nerve has an essential role in the appearance of cytoprotective mechanisms of atropine in GI tract. PMID- 10672994 TI - Detrimental effects of oestradiol on cysteamine-induced gastroduodenal ulceration in the female rat. AB - The actions of the female sex steroid, oestradiol on cysteamine-induced mucosal ulceration has been evaluated in female Wistar rats. Administration of cysteamine (400 mg x kg(-1), s.c.) provoked macroscopic gastroduodenal mucosal injury (assessed planimetrically) and an increase in microvascular permeability (assessed by the extravasation of radiolabeled albumin) in the stomach and duodenum, determined 24 h later. Ovariectomy (2 weeks before cysteamine) reduced gastroduodenal macroscopic injury, and albumin extravasation following cysteamine challenge. Administration of oestradiol (1-5 mg x kg(-1), as an i.m. depot 1 week before cysteamine) dose-dependently augmented gastric and duodenal macroscopic mucosal lesions and microvascular permeability provoked by cysteamine. These findings indicate that oestradiol can exacerbate gastroduodenal ulceration and microvascular injury. PMID- 10672995 TI - Effects of pectin-induced passive linkage of gastric H+ on the gastric acid secretion on the development of ethanol- and salicylate-induced gastric mucosal lesions in rats. AB - The aim of this study was to evaluate the effects of intragastrically given pectin-induced physicochemical properties and actions on active gastric acid secretion and on the development of ethanol- and aspirin-induced gastric mucosal lesions. The observations were carried out on CFY-strain rats, fasted for 24 h before the experiments with water ad libitum. The observations were carried out in two experimental series. A) The gastric mucosal lesions were produced by intragastrically given 96% ethanol or aspirin prepared with 0.2 M HCl. Different doses of pectin (100, 50 and 25 mg x kg(-1), respectively) were administered intragastrically 30 min before giving necrotizing agents. The number of gastric lesions was noted 1 h after the administration, while the severity of gastric mucosal lesions was scored by semi-quantitative scale. B) The effects of pectin were studied on the volume and H+ secretion of the stomach in 4-h pylorus-ligated rats. It has been found that: 1) the gastric mucosal lesions could be produced in 100% of rats by the application of both necrotizing agents. 2) Pectin in doses of 50-100 mg x kg(-1) increased the number of gastric mucosal lesions in both models, while no increase was produced by the application of 25-mg x kg(-1) dose. 3) The severity of mucosal lesions increased significantly after the administration of all doses of pectin. 4) The pectin-induced increase of gastric lesions (number) showed a dose-response effect. 5) The pectin produced a significant increase in the volume of gastric secretion and gastric H+ secretion. It has been concluded that: a) pectin-induced physicochemical changes are able to enhance the aggression to gastric mucosa produced by ethanol and aspirin; b) a positive correlation exists between the linkage of H+ to pectin and significant active metabolic response in the rat stomach; c) pectin alone stimulates the active metabolic process of the gastric H+ secretion. PMID- 10672996 TI - Pentadecapeptide BPC 157 attenuates gastric lesions induced by alloxan in rats and mice. AB - A diabetogenic alloxan regimen produced lesions in all stomachs of treated animals, either rats (200 mg x kg(-1) s.c.) or mice (400 mg x kg(-1) i.p.). In control animals, the lesions, when developed (i.e. 24 h following application), appear to be quite sustained, and consistently present also after 1 or 2 weeks. The application of the pentadecapeptide BPC 157 (10 microg or 10 ng x kg(-1) i.p. coadministered together with alloxan) would significantly attenuate these lesions' appearance. This beneficial effect seems to be present in either rats or mice and in either of the tested intervals. Importantly, the beneficial effect seems to be shared by both microgram and nanogram regimens. PMID- 10672997 TI - A behavioural study of the effect of pentadecapeptide BPC 157 in Parkinson's disease models in mice and gastric lesions induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydrophyridine. AB - The effect of a stomach pentadecapeptide, BPC 157, on Parkinson's disease in mice was investigated, along with its salutary activity on stomach lesions induced by parkinsongenic agents. Parkinsongenic agents, 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) (30.0 mg x kg(-1)b.w. i.p. once daily for 6d, and after 4d once 50.0 mg x kg(-1)b.w. i.p.) or reserpine (5.0 mg x kg(-1)b.w. i.p.) were applied i.p. BPC 157 (1.50 microg or 15.0 ng x kg(-1)b.w. i.p.) was applied 15 min before or alternatively 15 min after each MPTP administration. In reserpine studies, BPC 157 (10.0 microg or 10.0 ng x kg(-1)b.w. i.p.) was given either 15 min before reserpine or in the already established complete catalepsy 24 h thereafter. BPC 157 strongly improved the MPTP-impaired somatosensory orientation and reduced the MPTP-induced hyperactivity, and most importantly, MPTP-motor abnormalities (tremor, akinesia, catalepsy -otherwise very prominent in saline control), leading to almost complete abolition of otherwise regularly lethal course of MPTP treatment in controls. Likewise, in reserpine experiments, BPC 157 strongly prevented the development of otherwise very prominent catalepsy and when applied 24 h thereafter reversed the established catalepsy. In addition, a reduction of reserpine-hypothermy (BPC 157 pre-treatment) and reversal of further prominent temperature fall (BPC 157 post-treatment) have been consistently observed. Taking together these data, as the two most suitable animal models were consistently used and since the high effectiveness was demonstrated in pre- and post-treatment, microg and ng regimens, BPC 157 as an organoprotector should be further therapeutically investigated. Additionally, given in either regimen, pentadecapeptide BPC 157 strongly attenuated the stomach lesions in mice that otherwise consistently appeared in mice treated with the parkinsogenic neurotoxin MPTP. PMID- 10672998 TI - Characteristics of Chi distribution on different bacterial genomes. AB - The availability of full genome sequences provides the bases for analyzing global properties of the genetic text. For example, oligonucleotide sequences that are over- or underrepresented can be identified by taking into account the overall genome composition and organization. One of the most overrepresented oligonucleotides in Escherichia coli is the Chi site, an octanucleotide that stimulates DNA repair by homologous recombination. Here we analyze the genomic distribution of Chi in E. coli and in the three other bacteria where a Chi sequence has been identified; note that Chi is a different sequence in each organism. For each bacterial genome, Chi sequences are frequent, regularly distributed, and overrepresented. This suggests that selection for Chi may have occurred during evolution to favor efficient repair of a damaged chromosome. Other characteristics of Chi distribution are not conserved and might reflect specific features of DNA repair in each host. The different sequence and characteristics of Chi in each microorganism suggest that selection for Chi occurred independently in different bacteria. PMID- 10672999 TI - Trinucleotide repeats and other microsatellites in yeasts. AB - Microsatellites are direct tandem DNA repeats found in all genomes. A particular class of microsatellites, called trinucleotide repeats, is responsible for a number of neurological disorders in humans. We review here our current state of knowledge on trinucleotide repeat instability, and discuss the molecular mechanisms that may be involved in trinucleotide repeat expansions leading to fatal diseases in humans. We also present original data on microsatellite distribution in several microbial genomes, and on the use of microsatellites as physical markers to accurately and easily genotype yeast strains. PMID- 10673000 TI - DNA uptake signal sequences in naturally transformable bacteria. AB - The naturally transformable bacterium Haemophilus influenzae Rd contains 1471 copies of the DNA uptake signal sequence (USS) 5'-AAGTGCGGT in its genome. Neisseria meningitidis contains 1891 copies of the USS sequence 5'-GCCGTCTGAA. The USSs are often found in the base paired stem of transcription terminators. PMID- 10673001 TI - Occurrence and structure-function relationship of pentameric short sequence repeats in microbial genomes. AB - It is suggested that genomes found in any form of cellular life contain potentially size-variable repetitive DNA moieties. In eukaryotes, large proportions of the multi-chromosomal genome consist of various classes of repetitive DNA. Also in archaeal genomes, repetitive DNA is encountered and, as is the case for the eukaryotes as well, little or no function is at present attributable to most of it. For prokaryotes, elegant experiments have highlighted so-called slipped strand nucleotide mispairing (SSM) as a basic and causal mechanism, giving rise to repeat unit number variation at a distinct locus. Illegitimate base pairing in regions of repetitive DNA during replication, in association with defective DNA repair and enhanced nuclease susceptibility of replication intermediates, in the end gives rise to deletion or addition of repeat units. Prokaryotic short sequence repeats (SSRs) harbour arrays of short repeat units, between one and approximately 20 nucleotides in length. SSRs are involved in various mechanisms of microbial gene expression regulation. Promoter strength can be affected by altering the spacing between important structural domains as can the integrity of open reading frames. In the present communication the literature on microbial SSRs harbouring repeat units that are five nucleotides in length will be briefly reviewed. Examples of these SSRs with discrete functionality are encountered in bacterial species such as Haemophilus influenzae, Neisseria gonorrhoeae, and Pasteurella haemolytica. In addition, several of the currently known bacterial and archaeal whole genome sequences were scanned for the presence of novel examples of potential five-nucleotide SSRs (and others) in order to gather additional knowledge on the propensity and putative functions of this type of potential genetic switch. PMID- 10673003 TI - Super-integrons. AB - Integrons represent the primary mechanism for antibiotic resistance gene capture and dissemination among gram-negative bacteria. The recent finding of super integron (SI) structures in the genomes of several bacterial species has expanded their role in genome evolution. The Vibrio cholerae superintegron is gathered in a single chromosomal super-structure harbouring hundreds of gene cassettes. The encoded functions, when identifiable, are linked to adaptations extending beyond antibiotic resistance and pathogenicity. Comparison of the cassette contents of super-integrons from remote Vibrio species suggests that most of their cassettes are species-specific. Many bacterial species belonging to several distinct genera of the gamma- and beta-proteobacteria undoubtedly carry or show strong evidence for the presence of chromosomal SIs. If each bacterial species harbouring a SI has its own cassette pool, the resource in terms of gene cassette availability may be immense. PMID- 10673004 TI - Novel intergenic repeats of Escherichia coli K-12. AB - An online catalog of intergenic DNA repeat sequence elements is added to the EcoGene Escherichia coli K-12 genome sequence annotation and analysis project (bmb.med.miami.edu/EcoGene). A library of noncoding (intergenic) DNA sequences depleted of known intergenic repeat classes was searched for DNA sequence similarities to identify novel DNA repeat sequence classes. PMID- 10673002 TI - Short palindromic repetitive DNA elements in enterobacteria: a survey. AB - We present a survey of short palindromic repetitive elements in enterobacteria. Seven families are presented. Five were already known (RSA, IRU, 29-bp repeats, BIMEs and boxC), and their properties are updated; in particular, a new composite element is shown to include the formerly identified boxC repeats. Two repetitions, YPAL1 and YPAL2, found primarily in Yersinia, are described here for the first time. PMID- 10673005 TI - Large genomic sequence repetitions in bacteria: lessons from rRNA operons and Rhs elements. AB - The rrn operons and Rhs elements provide starkly contrasting examples of the evolution and interaction of large sequence repetitions in bacteria. Genomic sequencing of different species as well as comparative sequencing of independent isolates is providing provocative insights into previously obscure issues. PMID- 10673006 TI - IS elements as constituents of bacterial genomes. AB - We provide here an overview of our present understanding of the distribution of different insertion sequences (ISs) within bacterial genomes (both chromosomes and plasmids). This is at present fragmentary and a significant effort is needed in the analysis of the increasing number of genomes whose sequence has been determined. We also consider some of the properties of ISs which are important in their role of assembling, reassorting, and transmitting groups of genes. PMID- 10673007 TI - Paralogous genes encoding transport proteins in microbial genomes. AB - The largest superfamilies of prokaryotic genes encode transport proteins, but many transporters are encoded by orphan genes or those that comprise very small families. We have analyzed eighteen completely sequenced prokaryotic genomes for paralogous transport systems and have thereby identified 76 permease families. In this short review, we present and discuss the paralogues in some of these families and interpret the most prominent results, particularly those relevant to the largest permease superfamilies. PMID- 10673008 TI - DNA repeats and archaeal nucleosome positioning. AB - Archaeal histones, homologs of the eucaryal nucleosome core histones, have been identified in the Euryarchaeota. They assemble as tetramers with dsDNA to form archaeal nucleosomes that resemble the central structure of the eucaryal nucleosome formed by the histone (H3-H4)2 tetramer. Eucaryal and archaeal nucleosomes assemble preferentially on DNA molecules that best accommodate the severe bends found within these structures, and here we discuss the relationships between archaeal and eucaryal nucleosomes, repeating DNA sequences, and nucleosome positioning. PMID- 10673009 TI - Short repeats and IS elements in the extremely radiation-resistant bacterium Deinococcus radiodurans and comparison to other bacterial species. AB - Computer analysis of the complete genome of Deinococcus radiodurans R1 has shown that the number of insertion sequences (ISs) and small noncoding repeats (SNRs) it contains is very high, and comparable with those of Escherichia coli. IS elements and several families of SNRs are described, together with their possible function in the D. radiodurans genome. PMID- 10673010 TI - Functional and evolutionary roles of long repeats in prokaryotes. AB - Most recently published complete bacterial genomes have revealed unexpectedly high numbers of long strict repeats. In this article we discuss the various functional and evolutionary roles of these repeats, focusing in particular on their role in terms of genome stability, gene transfer, and antigenic variation. PMID- 10673011 TI - Repeated sequences in bacterial chromosomes and plasmids: a glimpse from sequenced genomes. AB - To gain insight into the extent of exact DNA repeats in sequenced bacterial genomes and their plasmids, we analyzed the collection of completely sequenced bacterial genomes available at GenBank using the program Miropeats. This program draws graphical representations of exact DNA repeats in whole genomes. In this work, we present maps showing the extent and type (inverted or direct) of exact DNA repeats longer than 300 bp for the whole collection. These repeats may participate in a variety of events relevant for bacterial genome plasticity, such as amplifications, deletions, inversions, and translocations (via homologous recombination), as well as transposition. Additionally, we review recent data showing that high-frequency architectural variations in genomic structure occur at both the interspecies and interstrain levels. PMID- 10673012 TI - Tandem repeats in complete bacterial genome sequences: sequence and structural analyses for comparative studies. AB - A series of complete bacterial genome sequences have recently become available and powerful methods have been developed for the identification of tandem repeats on a very large scale. It is thus possible to derive extensive comparative descriptions of such repeats at the level of complete genomes, as illustrated here for three different bacterial genomes: Escherichia coli, Haemophilus influenzae, and Mycobacterium tuberculosis. Such sequence analyses can be usefully complemented by structural characterisations of the repeats. PMID- 10673013 TI - Recognition of regulatory sites by genomic comparison. AB - Availability of complete bacterial genomes opens the way to the comparative approach to the recognition of transcription regulatory sites. Assumption of regulon conservation in conjunction with profile analysis provides two lines of independent evidence making it possible to make highly specific predictions. Recently this approach was used to analyze several regulons in eubacteria and archaebacteria. The present review covers recent advances in the comparative analysis of transcriptional regulation in prokaryotes and phylogenetic fingerprinting techniques in eukaryotes, and describes the emerging patterns of the evolution of regulatory systems. PMID- 10673014 TI - Three views of microbial genomes. AB - We describe here GenomeAtlases as a method for visualising three different aspects of complete microbial chromosomes: repeats, DNA structural characteristics, and base composition. We have applied this method to all publicly available genomes, and find a general strand preference of global repeats. The atlas for the Mycoplasma genitalium genome is presented as an example, and results from all three views are consistent with known characteristics of the genome. PMID- 10673015 TI - Promoter sequences and algorithmical methods for identifying them. AB - This paper presents a survey of currently available mathematical models and algorithmical methods for trying to identify promoter sequences. The methods concern both searching in a genome for a previously defined consensus and extracting a consensus from a set of sequences. Such methods were often tailored for either eukaryotes or prokaryotes although this does not preclude use of the same method for both types of organisms. The survey therefore covers all methods; however, emphasis is placed on prokaryotic promoter sequence identification. Illustrative applications of the main extracting algorithms are given for three bacteria. PMID- 10673016 TI - Synthesis, characterization, and platelet adhesion studies of novel aliphatic polyurethaneurea anionomers based on polydimethylsiloxane-polytetramethylene oxide soft segments. AB - Two novel aliphatic polyurethaneurea anionomers were synthesized based on polydimethylsiloxane (PDMS)-polytetramethylene oxide (PTMO) soft segments. The hard segments consisted of either 4,4'-methylene dicyclohexyl diisocyanate (H12MDI), sulfonic acid-containing diol and 1,4-butandiol (BD) or H12MDI, carboxylic acid-containing diol and BD. The nonionic counterpart chain extended with BD was prepared. In addition, the base nonionic polyurethaneurea containing a pure PDMS soft segment, which is denote H-D-BD, was also studied for comparison. The effects of soft segment type and ion incorporation on the physical properties, surface properties, and plateled adhesion are discussed. The ionic polyurethaneureas exhibited poor phase separation, a smaller fraction of PTMO present at the surface, and a smaller contact angle. On the other hand, it also showed a larger fraction of PDMS present at the surface and a higher water absorption value than its nonionic counterpart. H-D-BD had more phase-separated structure, a larger fraction of PDMS present at the surface, and larger contact angle but lower water absorption value than the PTMO-containing polyurethaneureas. The in vitro platelet adhesion experiments indicated that the ionic groups, especially for carboxylate, and surface enrichment PDMS soft segment could effectively inhibit platelet adhesion. PMID- 10673017 TI - Inhibition by heparin and derivatized dextrans of Staphylococcus epidermidis adhesion to in vitro fibronectin-coated or explanted polymer surfaces. AB - The ability of Staphylococcus aureus to recognize several extracellular matrix or plasma proteins (e.g., fibrinogen, fibronectin, and collagen) promotes bacterial attachment to artificial surfaces. Whereas most S. aureus clinical isolates elaborate a wide repertoire of bacterial surface receptors' called adhesins, exhibiting specific binding of individual host proteins, S. epidermidis is lacking most of such protein adhesins. To document the interactions between S. epidermidis and various surface-adsorbed proteins, we first compared promotion of bacterial attachment by seven purified human proteins immobilized onto poly(methyl methacrylate) (PMMA) coverslips. Only two of them, namely fibronectin and fibrinogen, exhibited adhesion-promoting activities. In the presence of native heparin or two functionalized dextrans (CMDBS for Carboxy Methyl, Benzylamide sulfonate/sulfate), a dose-dependent inhibition of S. epidermidis adhesion to fibronectin-coated, but not to fibrinogen-coated surfaces was observed. The inhibitory effects of each CMDBS were much stronger than that of native heparin. In contrast, a control highly negatively charged, dextran exclusively substituted with carboxy methyl groups exerted no inhibition on S. epidermidis adhesion. To evaluate how CMDBS could interfere with S. epidermidis attachment to coverslips coated in vivo with extracellular matrix components, we also tested PMMA surfaces retrieved from tissue cages subcutaneously implanted in guinea pigs. Each CMDBS, but not heparin, strongly inhibited S. epidermidis adhesion to explanted coverslips, even in the presence of tissue cage fluid. In conclusion, fibronectin plays an important role in promoting S. epidermidis attachment to implanted biomaterials. Furthermore, S. epidermidis adhesion to fibronectin-coated or implanted biomaterials can be efficiently blocked in vitro by CMDBS. PMID- 10673018 TI - Immobilization of invertase in conducting thiophene-capped poly(methylmethacrylate)/polypyrrole matrices. AB - Immobilization of invertase in thiophene-capped poly(methylmethacrylate)/polypyrrole matrices was achieved by constant potential electrolysis using different supporting electrolytes. Optimum reaction conditions such as substrate concentration, temperature, and pH for the enzyme electrodes were determined. The temperature and pH were found to be 60 degrees C and 4.8, respectively. The effect of supporting electrolyte on the enzyme activity revealed that SDS was the best in the immobilization procedure. Michaelis-Menten constant and the maximum reaction rate in PMMA/PPy matrices were of the order of that of pristine polypyrrole. However, in terms of repeated use, the copolymer matrices were superior to polypyrrole. PMID- 10673019 TI - Solubility control of enzymes by conjugation with stimulus-responsive polymers. AB - For development of bio-reactor enzymes stimuli-responsive polymers have been employed as immobilizing matrices. The stimuli-responsive polymers have been used for solubility control in water. Recently we first succeeded in the solubility control in organic media by conjugation with photo-responsive polymer. The polymer-conjugated enzymes can efficiently catalyze various chemical reactions in the soluble state and can be recovered by precipitation in response to the stimulation. In this review, the conjugation method and recent progress is described. PMID- 10673020 TI - Pulsatile peptide release from multi-layered hydrogel formulations consisting of poly(ethylene glycol)-grafted and ungrafted dextrans. AB - Multi-layered hydrogel formulations consisting of poly(ethylene glycol)-grafted dextran (PEG-g-Dex) and ungrafted Dex were investigated as a model of pulsatile drug release. In these formulations, it is considered that the grafted PEG domains act as a drug reservoir dispersed in the Dex matrix based on aqueous polymer two-phase systems. The formulations exhibited surface-controlled degradation by dextranase, and insulin release was observed in a pulsatile manner because of the multi-layered structure: PEG-g-Dex hydrogel layers containing insulin and insulin-free Dex hydrogel layers. Thus, it is suggested that the multi-layered hydrogel formulations using PEG-g-Dex and Dex are feasible for chronopharmacological drug delivery systems. PMID- 10673021 TI - Effects of long-term sub-lethal concentrations of dental monomers on THP-1 human monocytes. AB - Studies have shown that monomers from dental resins are acutely cytotoxic, but little is known of their long-term effects at sub-lethal concentrations. The current study determined the long-term effects of sub-lethal concentrations of TEGDMA (triethyleneglycol dimethacrylate) and Bis-GMA (bisphenol glycidylinethacrylate), two common dental monomers, on the in vitro cellular proliferation, succinic dehydrogenase activity, and total cellular protein production of monocytes. Human THP-1 monocytes were exposed to concentrations of 100, 200, and 400 micromol l(-1) of TEGDMA or 1, 5, and 25 micromol l(-1) Bis-GMA for 5 weeks. Controls received only vehicle solutions of ethanol. Each week cellular proliferation (hemocytometer), succinic dehydrogenase (SDH) activity (MTT) and total cellular protein (bicinchoninic acid) were assessed. The results were compared with ANOVA and Tukey intervals (alpha = 0.05). TEDGMA had no proliferative or cellular protein effects, but increased SDH activity 20-60% in week 1 (p < 0.05). SDH activity then decreased 40% in week 2, followed by a gradual increase of 30-40% over week 3-5 (p < 0.05). Bis-GMA reduced proliferation by 40-60% from 1-5 weeks exposure (p < 0.05). However, SDH activity and total protein per cell were not affected. There was some indication of increased SDH activity after 5 weeks (20-30%, p < 0.05). Sub-lethal concentrations of TEGDMA and Bis-GMA have significant long-term effects on monocytes at low-dose 5-week exposures in vitro. Each monomer acted differently. PMID- 10673022 TI - Effect of acetylation of biodegradable polyrotaxanes on its supramolecular dissociation via terminal ester hydrolysis. AB - Acetylation of biodegradable polyrotaxanes was examined to estimate the effect on its supramolecular dissociation via terminal ester hydrolysis. The biodegradable polyrotaxanes, in which many alpha-cyclodextrins (alpha-CD) are threaded onto a poly(ethylene glycol) chain capped with L-phenylalanine via ester linkages, were acetylated using acetic anhydride; alpha-CD release behavior was then characterized by in vitro hydrolysis. The degree of acetylation was changed by the concentration of acetic anhydride and the reaction time. The results of the in vitro hydrolysis indicate that the critical degree of acetylation to prolong supramolecular dissociation lies at around 30%. The terminal hydrolysis proceeded completely even with 100% of acetylation. These findings suggest that the hydrophobization of alpha-CDs in the polyrotaxane makes it possible to delay the time to complete the supramolecular dissociation. The hydrophobization of the polyrotaxane is of great importance for designing implantable materials that maintain their supramolecular structure until tissue regeneration with complete terminal hydrolysis. PMID- 10673023 TI - FTIR spectroscopic investigation and modeling of solute/polymer interactions in the hydrated state. AB - Attenuated total reflectance infrared spectroscopy was used to investigate possible interactions during transport of oxprenolol x HCl, bovine serum albumin, alpha-chymotrypsin, and fibrinogen through poly(acrylic acid) and its random copolymeric gels. Carbonyl and carboxylate ion peak shifts were used to identify drug/gel binding due to electrostatic and hydrogen bond interactions between the polymer carrier and the drugs tested. These findings were used to interpret the decrease in calculated diffusion coefficients of drugs diffusing through these gels and the associated hindering of drug transport. A model was developed to analyze this transport process as a function of the binding heat of the drug with the polymer. PMID- 10673024 TI - The mouse aldehyde oxidase gene: molecular cloning, chromosomal mapping and functional characterization of the 5'-flanking region. AB - In this article, we report on the chromosome mapping and molecular cloning of the genetic locus encoding the mouse molybdo-iron/sulfur-flavoprotein aldehyde oxidase. The aldehyde oxidase locus maps to mouse chromosome 1 band C1-C2, as determined by fluorescence in situ hybridization experiments conducted on metaphase chromosomes. The gene is approximately 83 kb long and consists of 35 exons. The exon/intron boundaries are perfectly conserved relative to the corresponding human homolog and almost completely conserved relative to the mouse xanthine oxidoreductase gene. This further supports the concept that the aldehyde oxidase and xanthine oxidoreductase loci evolved from the same ancestral precursor by a gene duplication event. The position of a major transcription start site was defined by primer extension and RNase mapping analysis. The 5' flanking region of the mouse aldehyde oxidase gene contains a functional and orientation-dependent promoter as well as several putative binding sites for known cell-specific and general transcription factors. Deletion analysis of the 5'-flanking region defines an approximately 470 bp DNA stretch which is necessary and sufficient for the transcription of the mouse aldehyde oxidase gene. PMID- 10673025 TI - A second-site mutation at glutamate-257 that restores the function of the mutant yeast ribosomal protein L5 containing lysine-270,271-->arginine. AB - A genetic approach was used to identify interacting regions of yeast ribosomal protein L5 (also known as L1, L1a, or YL3). Previous studies from our laboratory showed that residues K270 and K271 in protein L5 are essential for its function. The mutant L5 protein in which both residues were replaced by arginine residues (K270,271R) exhibited about 80% RNA binding capability compared to the wild-type and the mutant protein was assembled into the 60S ribosomal subunits in vivo. The yeast strain expressing this mutant protein in a homozygous form was lethal (Biochim. Biophys. Acta 1308 (1996) 133-141). In the present study, this non functional mutant was used to select intragenic suppressors. A spontaneous, intragenic suppressor which contained an E257K substitution (in addition to the primary mutations) was identified. The suppressor protein bound about 60% of yeast 5S rRNA in vitro compared to the wild-type. To gain more insight into the nature of the intragenic suppressor, additional mutant proteins in which E257 was substituted by a variety of amino acids were produced by site-directed mutagenesis. The ability of each mutant protein to bind yeast 5S rRNA in vitro and to suppress the lethal effect of the double K270,271 mutation in vivo were examined. Results suggest communication between two non-contiguous domains on protein L5 and that several factors, such as electrostatic interaction and hydrogen bonding are likely to play a role in this global communication. Mutation studies on E257 alone also reveal that substitutions of this residue in L5 protein could affect cell growth under specified conditions, but a variety of changes could be tolerated without serious deleterious effects. We propose a working model in which E257 is located in a loop and the dynamic as well as the flexibility of this loop is important for L5 function. PMID- 10673026 TI - The influence of the exocyclic pyrimidine 5-methyl group on DNAse I cleavage and sequence recognition by drugs. AB - Incorporation of modified nucleotides into DNA, using the PCR, has allowed us to probe the influence that the exocyclic 5-methyl group of pyrimidines has on DNAse I cleavage and sequence recognition by drugs. The results show that removal of the methyl group from the major groove, made possible by substituting uridine for thymidine, allows DNAse I to cleave more readily at AT-rich regions compared to normal DNA. By contrast, addition of an extra methyl group, contrived by substituting 5-methylcytidine for normal cytidine, allows DNAse I to cleave more readily at GC-rich regions compared to normal DNA. In the cutting pattern of DNA containing both uridine and 5-methyl cytosine, we find the cleavage characteristics of both the single-substituted DNA species combined. Thus, the presence or absence of the exocyclic 5-methyl group in the major groove has a strong influence on the relative intensity of cleavage of phosphodiester bonds by DNAse I. These nucleotide substitutions can also influence the sequence-selective binding of drugs to DNA. Whereas removal of the methyl group (replacement of T with U) generally has little effect on sequence recognition by a variety of drugs, addition of a methyl group (replacement of C with M) generates new binding sites for some intercalators, namely daunomycin, DACA and SN16713. PMID- 10673027 TI - Mapping of eukaryotic DNA topoisomerase I catalyzed cleavage without concomitant religation in the vicinity of DNA structural anomalies. AB - Sensitive sites for covalent trapping of eukaryotic topoisomerase I at DNA structural anomalies were mapped by a new method using purified enzyme and defined DNA substrates. To insure that the obtained topoisomerase I trapping patterns were not influenced by DNA sequence variations, a single DNA imperfection was placed centrally within a homonucleotide track. Mapping of topoisomerase I-mediated irreversible cleavage sites on homopolymeric DNA substrates containing mismatches showed trapping of the enzyme in several positions in close vicinity of the DNA imperfection, with a strong preference for the 5' junction between the duplex DNA and the base-pairing anomaly. On homopolymeric DNA substrates containing a nick, sites of topoisomerase I-mediated cleavage on the intact strand were located just opposite to the nick and from one to ten nucleotides 5' to the nick. Sites of enzyme-mediated cleavage next to a nick and an immobile single-stranded branch were located 5' to the strand interruption in distances of two to six nucleotides and two to ten nucleotides, respectively. Taken together these findings suggest that covalent trapping of topoisomerase I proceeds at positions adjacent to mismatches, nicks and single stranded branches, where the cleavage reaction is allowed and the ensuing ligation reaction prevented. In principle, the developed interference method might be of general utility to define topoisomerase-DNA interactions relative to different types of structural anomalies. PMID- 10673028 TI - Tumor promotion resistant cells are deficient in AP-1 DNA binding, JunD DNA binding and JunD expression and form different AP-1-DNA complexes than promotion sensitive cells. AB - The JB6 cell culture model is used to identify molecular determinants of susceptibility to the promotion of neoplastic transformation. Clonal variants susceptible to transformation ('P+' cells) form numerous anchorage-independent colonies in soft agar upon treatment with the phorbol ester tumor promoter TPA, whereas resistant variants ('P-' cells) do not. We now report that there is significantly less binding of activator protein-1 (AP-1) to its DNA binding site in P- cells than in P+ cells. Gel supershift assays were performed to detect association of all seven AP-1 family members with their DNA binding site in TPA treated and -untreated P+ and P- cells. Significantly lower DNA binding and protein expression of JunD were detected in P- cells than in P+ cells. c-Jun was detected in P+, but not P-, AP-1-DNA complexes, and c-Fos was detected in P-, but not P+, AP-1-DNA complexes. These and other phenotype-specific differences in abundance and composition of AP-1-DNA complexes may play a role in the resistance of P- cells to tumor promoter-induced transformation. PMID- 10673029 TI - Characterization of the Azoarcus ribozyme: tight binding to guanosine and substrate by an unusually small group I ribozyme. AB - We report novel chemical properties of the ribozyme derived from the smallest group I intron (subgroup IC3) that comes from the pre-tRNA(Ile) of the bacterium Azoarcus sp. BH72. Despite the small size of the Azoarcus ribozyme (195 nucleotides (nt)), it binds tightly to the guanosine nucleophile (Kd = 15 +/- 3 microM) and exhibits activity at high temperatures (approximately 60-70 degrees C). These features may be due to the two GA3 tetraloop interactions postulated in the intron and the high GC content of the secondary structure. The second order rate constant for the Azoarcus ribozyme, ((k(cat)/Km)S = 8.4 +/- 2.1 x 10(-5) M( 1) min(-1)) is close to that found for the related ribozyme derived from the pre tRNA(Ile) of the cyanobacterium Anabaena PCC7120. pH dependence studies and kinetic analyses of deoxy-substituted substrates suggest that the chemical cleavage step is the rate-determining process in the Azoarcus ribozyme. This may be due to the short 3-nt guide sequence-substrate pairing present in the Azoarcus ribozyme. Finally, the Azoarcus ribozyme shares features conserved in other group I ribozymes including the pH profile, the stereospecificity for the Rp phosphorothioate at the cleavage site and the 1000-fold decrease in cleavage rate with a deoxyribonucleoside leaving group. PMID- 10673030 TI - Scanning gene expression during neuronal cell death evoked by nerve growth factor depletion. AB - Depletion of nerve growth factor (NGF) from differentiated, neuronal PC12 cells causes a form of programmed cell death that stems from the attenuation of NGF receptor signaling and the resultant expression of certain genes required for cell death. To better understand the associated molecular events, we surveyed the changes in gene expression in PC6-3 cells, a subline of PC12, caused by depletion of NGF. Using restriction landmark cDNA scanning, we assessed the expression patterns of as many as 15,000 gene species, and 30 genes were isolated whose expression was altered in the absence of NGF. Of the 20 genes up-regulated in the absence of NGF, including transcription factor LRF-1/ATF3, most were also up regulated during the programmed death of cortical neurons caused by Ca2+ ionophore. Their function may thus be a general feature of programmed neuronal cell death. In contrast, with one exception, expression of down-regulated genes was NGF-dependent and therefore diminished in the absence of NGF but unaffected by Ca2+ ionophore. These findings confirm that global investigation of the features of up- and down-regulated genes should add substantially to our understanding of the regulation of programmed neuronal cell death and the mechanisms involved. PMID- 10673031 TI - Identification of novel isoforms of human RAD52. AB - In yeast, RAD52 has been shown to be essential for homologous recombination of DNA and to be involved in the repair of double-stranded DNA breaks. Recently, the human homologue of yeast RAD52, a 418-amino-acid protein, has been identified. In this study, we report three different isoforms of human RAD52 isolated from brain and testis cDNA libraries. cDNAs of these isoforms contain distinct insertions and encode truncated proteins due to translational frame-shifts. The three isoforms consist of 226-, 139-, and 118-amino-acid residues, and are designated as RAD52beta, gamma, and delta, respectively. The original RAD52 is termed as RAD52alpha in this paper. Messages of these isoforms have been detected in various human tissues. We found that the RAD52 isoforms were unable to interact with RAD52alpha because of partial defect of the self-interaction domain. Furthermore, like RAD52alpha, the isoforms have been shown to bind to both single stranded and double-stranded DNA. These results suggest that RAD52beta, gamma, and delta might affect RAD52alpha function through their DNA-binding property and their inability to bind to RAD52alpha. Thus, these isoforms might act as dominant negative mutants or negative regulators of RAD52alpha. PMID- 10673032 TI - Structure and genomic organization of porcine RACK1 gene. AB - The cDNA encoding porcine RACK1 protein was isolated from porcine spleen cDNA library. The deduced protein sequence of porcine RACK1 cDNA shows that it contains 317 amino acid residues, and shares nearly 100% identity with its vertebrate counterparts. Noticeably, the RACK1 protein was differentially expressed in various porcine tissues. High expression of RACK1 protein was observed in the tissues including thymus, pituitary, spleen and liver, whereas there was no detectable expression in muscle. The genomic DNA of porcine RACK1 with approximate 7.5 kb was constructed by both polymerase chain reaction amplification and genomic library screening. It consists of eight exons intervened by seven introns, and most of the intron/exon splice sites conform to the GT/AG rule. The promoter region contains functional serum response element, YY1-like binding site and AP1 site, which is supported by the finding that the expression of RACK1 gene in cultured porcine ST cells has a serum response as well as a TPA response. PMID- 10673033 TI - Three structurally and functionally conserved Hlx genes in zebrafish. AB - For the Hlx class, which includes homeodomains (HD) that are similar to Drosophila H2.0, few members have been identified in vertebrates. In this report, we describe three zebrafish genes, hlx1, hlx2 and hlx3, related to the murine Dbx genes. The proteins encoded by hlx1 and hlx2 have about the same sequence identity to Dbx1 (approximately 60%), suggesting that they derive from a duplication in the fish lineage. This is supported by similarities in the embryonic expression patterns and promoter sequence conservation. The zebrafish Hlx3 protein is related to murine Dbx2, but it is apparently too diverged to be orthologous. Our phylogenetic analysis of all the known HD sequences of the Hlx class also shows that it can be divided into at least two distinct families. All the Dbx-like genes have similar expression in the embryonic nervous system. However, the initial expression patterns of the zebrafish hlx genes are quite unique, suggesting that some functional divergence has occurred between fish and mammals. PMID- 10673034 TI - Quantitative analysis reveals differential expression of mucin (MUC2) and intestinal trefoil factor mRNAs along the longitudinal axis of rat intestine. AB - MUC2 and intestinal trefoil factor (ITF) are considered to have important roles in intestinal mucosal protection and epithelial repair. In order to investigate whether these genes are co-ordinately expressed, we have used competitive reverse transcription-polymerase chain reaction assays to measure MUC2 and ITF mRNA levels in human intestinal cell lines and along the longitudinal axis of rat intestine. ITF mRNA was expressed in several intestinal cell lines. However, MUC2 mRNA was detected only in LS174T cells where it was present at approx. 25-fold lower levels than the ITF transcript. In contrast, in rat intestinal tissues, MUC2 mRNA levels were generally higher than ITF mRNA levels. The levels of both transcripts increased markedly during postnatal development. In adult rats, the expression patterns of MUC2 and ITF mRNAs along the longitudinal axis of the small intestine were similar, with lowest levels in the proximal duodenum and relatively constant levels in the other regions assayed. In contrast, the expression patterns of MUC2 and ITF in different regions of the large intestine showed a marked divergence. Our results strongly suggest that expression of the MUC2 and ITF genes is not coordinately regulated in intestinal cells. PMID- 10673035 TI - Structure of the Xenopus laevis TFF-gene xP4.1, differentially expressed to its duplicated homolog xP4.2. AB - TFF-peptides (formerly P-domain peptides, trefoil factors) represent major secretory products of mucous epithelia in mammals and amphibia. The nucleotide sequence of a large portion of a gene encoding the TFF-peptide xP4.1 from Xenopus laevis and its genomic organization were determined in the present study. The peptide xP4.1 containing four TFF-domains is thought to represent the functional frog homolog of human TFF2 (formerly hSP). The xP4.1 gene analyzed spans a region of about 7 kb and consists of six exons. Each TFF-domain is encoded by a single exon flanked by type 1 introns typical of shuffled modules. The 5'-upstream region contains a TATA-box, and potential binding sites for hepatocyte nuclear factor 3 and AP-1. Furthermore, the cDNA sequence of a transcript named xP4.2 with 91% similarity to xP4.1 is presented. RT-PCR analysis revealed that xP4.1 and xP4.2 genes are differentially expressed. xP4.1 transcripts are detectable only in the stomach, but not in the esophagus, whereas xP4.2 transcripts are found both in the esophagus and in the stomach with a descending gradient from fundus to antrum. PMID- 10673036 TI - Activation of Smad1-mediated transcription by p300/CBP. AB - Smad1 is the intracellular effector of bone morphogenetic proteins (BMPs), a growth factor family well known to stimulate bone and cartilage formation. Smad1 becomes phosphorylated by the cognate BMP transmembrane receptor protein kinases, associates with the common mediator Smad4 and translocates to the nucleus where it is involved in regulation of gene transcription. In this report we demonstrate that Smad1 interacts with the paralogous coactivators p300 and CBP both in vitro and in vivo. The N- and C-termini of Smad1 negatively interfere with binding to p300/CBP, and the C-terminal half of Smad1 harbors two interaction domains both binding to the same region near the C-terminus of p300/CBP. We show that Smad1 as well as a Gal4 fusion with the C-terminal half of Smad1 stimulate gene transcription in a cooperative fashion with p300/CBP. Phosphorylation of Smad1 enhances its binding to CBP and thereby further stimulates Smad1-dependent transcription. These results provide a mechanism how phosphorylated Smad1 regulates gene activity by interaction with p300/CBP, and factors associated with these bridging coactivators. PMID- 10673037 TI - Molecular cloning and characterization of the mouse histone deacetylase 1 gene: integration of a retrovirus in 129SV mice. AB - Reversible histone acetylation plays an important role for chromatin structure and gene expression. The acetylation state of core histones is controlled by histone acetyltransferases and histone deacetylases. Here we report the cloning and characterization of the mouse histone deacetylase 1 (HDAC1) gene. The mouse genome contains several HDAC1-related structures representing the HDAC1 gene and at least three pseudogenes. The HDAC1 gene comprises 14 exons ranging from 49 to 539 bp. Interestingly the murine HDAC1 gene strongly resembles the previously published mouse HDAC2 gene (Zeng et al., J. Biol. Chem. 273 (1998) 28921-28930). The sizes of ten of the 14 exons are identical for both genes and the splicing sites for 11 introns align in identical positions suggesting a gene duplication event. The HDAC1 gene is located only 128 bp downstream from the MARCKS-related protein (MRP) gene in a tail-to-tail orientation. The murine MRP gene was previously mapped to a conserved gene cluster on chromosome 4 sharing linkage homology to human chromosome 1p32-36. The genes for HDAC1 and MRP are co expressed in a variety of cell types. In the genome of 129SV mice the largest intervening sequence of the HDAC1 gene, intron 3, harbors a complete copy of the endogenous retrovirus MuERV-L. In contrast the HDAC1 gene in other mouse strains such as C57B16, C3H/An and C-RY lacks the retrovirus. Our study provides useful tools for future targeted gene disruption studies. PMID- 10673038 TI - The stem hairpin loop structure of p2Sp1 RNA is required for RNA-cleaving activity. AB - We studied the hairpin-loop structure of an RNA fragment (GUUUCGUACAAAC) (R13) with the sequence corresponding to the self-cleavage domain in the precursor of an RNA molecule from bacteriophage T4-infected Escherichia coli cells (p2Sp1 RNA). In order to determine the influence of the hairpin-loop structure on these sequence-specific cleavage reactions, we have synthesized oligoribonucleotides containing hairpin-loop, double-helical stem-loop, and single-stranded RNA structures. The cleavage was affected by the hairpin-loop structure. Furthermore, the helix-stem, which retains the thermodynamically extrastable stem hairpin-loop structures, is also important for the cleavage activity. However, the thermodynamically extrastable helix-stem structure reduced the cleavage activity of the adjacent UA and CA sequences at the helix-stem site. For the cleavage reactions of the RNA cleavage products, the R6 (ACAAAC), R7 (GUUUCGU), and R9 (GUUUCGUAC) mers from the parent RNA, R13 (GUUUCGUACAAAC), a very slight amount of cleavage product (2%) from the RNA 9 was observed, but no reaction occurred for the R6 and R7. We also describe the influences of the sequences (UA and CA) on the cleavage activity. PMID- 10673039 TI - Characterization of the 5'-flanking region of the gene encoding bovine adenylate kinase isozyme 3. AB - We have characterized the 5'-flanking region of the gene encoding bovine adenylate kinase isozyme 3 (AK3). S1 mapping analysis revealed multiple transcription start points in the bovine AK3 gene. The promoter activities were tested in HeLa cells using the chloramphenicol acetyltransferase (CAT) gene as a reporter. The CAT analysis showed that the basal promoter sequence was located within the region from -189 to +228. In the presence of short DNA fragments of the 5'-flanking region as competitors, the transcriptional activity of the bovine AK3 promoter changed depending on the fragments used. The results identified the basal regulatory elements in the proximal promoter region. PMID- 10673040 TI - Characterization of the full-length sequences of phospholipase A2 induced during flower development. AB - The suppression subtractive hybridization (SSH) method was used to isolate developmentally regulated genes during carnation flower maturation. Carnation flower maturation-related clones obtained by the SSH were serially assigned as CFMI (carnation flower maturation-induced) clones. Northern blot analysis showed that several CFMI clones were differentially expressed during flower development. One of the clones, CFMI-3, showed similarity to various animal secretory phospholipases A2 (PLA2). Since little is known about PLA2 gene sequence in plant species, the CFMI-3 clone was selected for further characterization by sequence analysis. Full sequence analysis reveals that the CFMI-3 contains a Ca2+ binding domain, a PLA2 active site, and 12 conserved Cys residues, which is a distinct characteristic of PLA2. Amino acid sequence alignment of CFMI-3 to various putative plant PLA2 confirmed that the CFMI-3 cDNA is the full-length putative PLA2 cDNA identified in plant species. PMID- 10673041 TI - Molecular cloning of two bovine aquaporin-4 cDNA isoforms and their expression in brain endothelial cells. AB - Two cDNA isoforms of bovine aquaporin-4 (bAQP4-A and bAQP4-B) were newly isolated. Sequence analysis of both cDNAs revealed open reading frames of 972 (bAQP4-A) and 906 nucleotides (bAQP4-B) with deduced proteins of 323 (bAQP4-A) and 301 amino acid residues (bAQP4-B). Partial 5'-genomic sequence analysis showed that the 5'-noncoding sequences specific to bAQP4-A and -B transcripts were contained in distinct exons, exon 0 for bAQP4-A and new exon X for bAQP4-B. RNase protection assay demonstrated the definite expression of both isoforms in bovine brain. The deduced amino acid sequence of bAQP4-A was highly homologous to the human (97%), rat (95%), and mouse (93%) AQP4. Reverse transcription-PCR detected the expression of AQP4 mRNAs in bovine brain endothelial cells as well as in a variety of bovine organs such as brain, lung, spleen, and kidney. Northern blot analysis indicated that a 6.0 kb message is predominantly expressed in bovine brain and lung. PMID- 10673042 TI - Cloning and characterization of the unusual cyclin gene from an amphidiploid of Nicotiana glauca-Nicotiana langsdorffii hybrid. AB - Cyclins play an important role in the regulation of cell cycle progression in eukaryotic cells. As an aid to understanding the molecular nature of unregulated cell proliferation, a cDNA clone encoding a cyclin gene, GTcyc, was identified from genetic tumors. The clone contained 1095 bp including a 24 base poly(A) tail. GTcyc is an unusual cyclin gene, distantly related to mammalian cyclin D genes having 21-25% identity within the cyclin box. Northern blots showed that the genetic tumors express high levels of GTcyc relative to non-tumor hybrid tissues. Southern analysis suggests that GTcyc may be contained one or two families in genetic tumors. PMID- 10673044 TI - Cloning and characterisation of the rpoS gene from plant growth-promoting Pseudomonas putida WCS358: RpoS is not involved in siderophore and homoserine lactone production. AB - The rpoS gene which encodes a stationary phase sigma factor has been identified and characterised from the rhizosphere-colonising plant growth-promoting Pseudomonas putida strain WCS358. The predicted protein sequence has extensive homologies with the RpoS proteins form other bacteria, in particular with the RpoS sigma factors of the fluorescent pseudomonads. A genomic transposon insertion in the rpoS gene was constructed, these mutants were analysed for their ability to produce siderophore (iron-transport agent) and the autoinducer quorum sensing molecules called homoserine lactones (AHL). It was determined that RpoS was not involved in the regulation of siderophore and AHL production, synthesis of these molecules is important for gene expression at stationary phase. P. putida WCS358 produces at least three different AHL molecules. PMID- 10673043 TI - Cloning and characterization of a novel human gene encoding a zinc finger protein with 25 fingers. AB - This study reports cloning and characterization of a human cDNA encoding a novel human zinc finger protein, ZFD25. ZFD25 cDNA is 6118 bp long and has an open reading frame of 2352 bp that encodes a 783 amino acid protein with 25 C2H2-type zinc fingers. The ZFD25 cDNA also contains a region with high sequence similarity to the Kruppel-associated box A and B domain in the 5'-untranslated region, suggesting that ZFD25 belongs to the Kruppel-associated box zinc finger protein family. The ZFD25 gene was localized to chromosome 7q11.2. Northern blot analysis showed that ZFD25 was expressed in a wide range of human organs. In cultured endothelial cells, the mRNA level was decreased upon serum starvation. PMID- 10673045 TI - Cloning and expression profile of mouse and human genes, Rnf11/RNF11, encoding a novel RING-H2 finger protein. AB - The RING finger (C3HC4-type zinc finger) is a variant zinc finger motif presents in a new family of proteins. A new member of the RING finger family was identified and its cDNA structures were determined in human and mouse. The predicted protein consisting of a 144 amino acid residues is very conservative between the two species and contains a canonical RING-H2 finger motif (C3H2C2) at the carboxyl-terminal region. The genes were designated as RNF11/Rnf11 for RING finger protein 11. A single 2.4-kb transcript of mouse Rnf11 was ubiquitously expressed in various fetal and adult mouse tissues by the Northern blot analysis. The human RNF11 gene was mapped on chromosome 1p31-p32 region, where frequent alterations have been observed in T-cell acute lymphoblastic leukemia. PMID- 10673046 TI - Characterization of a new ATP-binding cassette transporter in Trypanosoma cruzi associated to a L1Tc retrotransposon. AB - We have characterized the tcpgp1-like gene of Trypanosoma cruzi, a new ATP binding cassette (ABC) transporter. tcpgp1 codes for a 1035 amino acid protein with a considerable homology to LtpgpA of Leishmania. Tcpgp1 lacks the conserved sequences corresponding to the second nucleotide-binding domain of other ABC transporters due to the insertion of the L1Tc non-LTR retrotransposon. PMID- 10673047 TI - A novel TNF-inducible message with putative growth suppressor function. AB - We report the nucleotide sequence of a novel cDNA and TNF-induced expression of the corresponding message (mRNA) in human fibroblast cells. This message is also expressed in certain human tumor cell lines and is over-expressed in a colon cancer cell line (HT-29). NIH3T3 cells transfected with the antisense construct of the 5'-region of this novel cDNA formed 20-fold more colonies in culture compared to cells transfected with a sense construct of the same region or the sense and the antisense constructs of the central region of this cDNA. This observation suggests a possible growth suppressor function for the gene represented by this cDNA. PMID- 10673048 TI - Calcyclin is differentially expressed in rat testicular cells. AB - Immature rat Sertoli cells aggregate and form tubule-like structures when cultured on a monolayer of peritubular myoid cells. In this study, differential gene expression of monocultures and direct cocultures of peritubular cells and Sertoli cells were examined. One of the cDNA clones isolated showed high homology to calcyclin and a microvascular differentiation gene, CEC5, which was reported to be highly homologous to CASK, a membrane-associated guanylate kinase homolog. Sequencing and mRNA analysis of rat calcyclin demonstrated that the gene was differentially expressed and was found only in peritubular cells and cocultures with increased levels. In contrast, CASK was expressed by Sertoli cells, peritubular cells, and cocultures, whereas CEC5 was never found in the testicular somatic cells. Our findings point to a paracrine regulation of calcyclin expression in testicular peritubular fibroblasts which seems to be related to tubular growth. PMID- 10673049 TI - Molecular characterisation of In51, a class 1 integron containing a novel aminoglycoside adenylyltransferase gene cassette, aadA6, in Pseudomonas aeruginosa. AB - Polymerase chain reaction-amplification and subsequent sequencing of the variable region of a novel integron, In51, from Pseudomonas aeruginosa revealed the presence of a novel aminoglycoside adenylyltransferase gene, aadA6, together with an open reading frame of unknown function, orfD. AADA6 enzyme has only 75% amino acid identity with AADA1 and is able to confer high level resistance to streptomycin and spectinomycin in Escherichia coli. PMID- 10673050 TI - Cloning, expression and characterization of a novel human Ras-related protein that is regulated by glucocorticoid hormone. AB - Ras proteins are a family of guanine nucleotide (GDP and GTP)-binding proteins that play central roles in essential signal transduction pathways. We have isolated in a yeast two-hybrid screen a human cDNA encoding a new protein that is highly homologous (98% identical at the protein level) to mouse DexRas1, a member of the Ras superfamily. The human DexRas1 is expressed in a variety of tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas, with the strongest expression in the heart. Using human fibrosarcoma HT 1080 cells as a model system, we show that the expression of human DexRas1 is stimulated by dexamethasone, suggesting a role of human DexRas1 in dexamethasone induced alterations in cell morphology, growth and cell-extracellular matrix interactions. PMID- 10673051 TI - Genetic analysis of the gene cluster responsible for synthesis of serotype e specific polysaccharide antigen in Actinobacillus actinomycetemcomitans. AB - A gene cluster associated with the biosynthesis of the serotype e-specific polysaccharide antigen (SPA) of Actinobacillus actinomycetemcomitans IDH1705 belonging to serotype e was cloned and sequenced. This cluster consisted of 18 open reading frames. Escherichia coli produced the polysaccharide that reacts with the serotype e-specific antiserum when transformed with a plasmid containing the cluster. Comparing the structure of the gene cluster with similar clusters from A. actinomycetemcomitans strains Y4 (serotype b) and NCTC9710 (serotype c) revealed that a 5.3-kb region containing the distal half of one gene and two entire genes in the cluster from strain IDH1705 replaced a 6.2-kb region containing eight genes in the cluster from strain Y4, and a 4.7-kb region containing four genes in the cluster from strain NCTC9710. These results suggest that this region is essential to the antigenic specificity of serotype e A. actinomycetemcomitans. PMID- 10673052 TI - cDNA cloning and gene expression of three small GTP-binding proteins in defense response of chickpea. AB - The cDNA clones encoding rab type (INR134 and ELR19) and rac type (ELR26) small GTP-binding proteins were isolated from Ascochyta rabiei-inoculated chickpea leaves and the elicitor-treated cell cultures. Rac type ELR26 showed enhanced expression in inoculated leaves indicating correlation with the defense response. PMID- 10673053 TI - Characterisation and quantification of earthworm cyclophilins: identification of invariant and heavy metal responsive isoforms. AB - We report the identification of two earthworm cyclophilin genes, which resemble the cytosolic cyclophilin-A and the signal sequence containing cyclophilin-B. Using fully quantitative PCR we were able to assess the transcript regulation of both cyclophilin isoforms, as well as a further independent control gene (actin), during exposure to heavy metals. Whilst the expression of cyclophilin-B and actin remained exceptionally constant, cyclophilin-A was up-regulated 38-fold. This intriguing observation has profound implications regarding cyclophilin's use as an invariant control and highlights the fact that it is essential to treat cyclophilin isoforms as separate entities, rather than one functional unit. PMID- 10673054 TI - Experimental trials with a thermostable Newcastle disease virus (strain I2) in commercial and village chickens in Tanzania. AB - Antibody responses in indigenous village and commercial chickens vaccinated with 12 thermostable Newcastle disease (ND) vaccine and protection levels against challenge with a virulent field isolate were determined. The antibody response of village chickens vaccinated by eye drop revealed that 30, 60 and 90 days after primary vaccination, the mean log2 HI titres were 6.1, 5.4 and 3.6, respectively, whereas for commercial chickens, the antibody response after 14, 30 and 90 days were 8.2, 5.1 and 4.2, respectively. Village chickens vaccinated orally via drinking water had mean log2 HI titres of 3.4 after 30 days. After booster vaccination, the mean HI titre was 5.4 and 3.3 after 30 and 60 days post secondary vaccination (i.e. 60 and 90 days after primary vaccination). Antibody response of mean log2 HI titres of 2.6 was recorded 30 days after primary vaccination orally through food; 30 and 60 days after secondary vaccination (i.e. 60 and 90 days after primary vaccination), mean log2 HI titres were 5.3 and 3.2, respectively. All commercial and village chickens vaccinated by eye drop survived the challenge trial whereas village chickens vaccinated through drinking water and food had protection levels of 80% and 60% 30 days after primary vaccination, respectively. However, 30 days after booster vaccination, the protection level was 100%. At 60 days after secondary vaccination, the protection level dropped again to 80% for chickens vaccinated orally. All control chickens used in the challenge trials developed clinical ND and died 3-5 days after inoculation with the virulent virus. Supported by laboratory findings, I2 strain of NDV seemed to be avirulent, immunogenic and highly protective against virulent isolates of NDV. It may be a suitable vaccine to use in village chickens to vaccinate them against ND in rural areas. PMID- 10673055 TI - Incidence risk and clinical features of retention of foetal membranes in ewes in 28 flocks in southern Greece. AB - In a field investigation of 28 flocks in southern Greece, 7660 lambings were monitored. Retention of foetal membranes (defined as failure to expel the foetal membranes within 6 h of lambing the last lamb) was recorded in 92 ewes. The median within-flock incidence risk was 1.25% (range: 0-1.9%). The heterogeneity of the risk among flocks was not significant (p = 0.99); no correlation was found between the incidence risk and the flock size (r(sp) = 0.33, p = 0.09). Overall, the median duration of retention was 72 h (range: 9-288 h); it did not differ among flocks (p = 0.89) and was not correlated with flock size (r(sp) = 0.24, p = 0.27). During the initial stage of retention the membranes were fleshy and humid; the cotyledons were thick and congested. Progressively, the membranes became thin, dry and stringy; the cotyledons shrunk and were pale. Finally the membranes dropped as a mass. In ewes with retention for >12 h, a variety of accompanying signs was recorded: straining (in 22% of the ewes with retention), vulval oedema and reddening (in 16%), anorexia (in 13%), recumbency (in 13%) and increased temperature (in 12%) were the most frequent ones. Overall, the median clinical score of the disorder was '2' (range: '1'-'5'); it did not differ among flocks (p = 0.98) and was not correlated with flock size (r(sp) = 0.26, p = 0.25). In 4% of the ewes with retention for <4 days accompanying signs were recorded (median clinical score: '2'), whilst in 80% of the ewes with retention for > or =4 days accompanying signs were recorded (median clinical score: '3'). This difference in the prevalence of clinical signs was significant (p < 0.0001). PMID- 10673056 TI - Parents' predicted transmitting abilities are not associated with culling prior to second lactation of Michigan, USA dairy cows. AB - The objective of this study was to test the association between parents' predicted transmitting abilities (PTAs) for milk, fat and protein and subsequent culling of heifers during rearing or first lactation. The data from 1992 to 1996 consisted of the culling outcome of 5619 Michigan Holstein heifers during rearing or first lactation, and the heifers' parents' PTAs from the period in which the heifers were born. Fixed-effect probit models (both dichotomous and ordered) estimated the relationships between culling and parents' PTAs. Herd effect was modeled as a fixed-effect. There was no association between PTAs of milk, fat, and protein for each heifer's parents and subsequent culling of the heifer during rearing or first lactation. Weak evidence for the necessity of modeling the herd effect as fixed was present. The lack of association between parents' PTAs and reason-specific culling (voluntary versus involuntary)--coupled with the counter intuitive effects of parents' PTAs on the probability of being culled for each of the reasons--raised questions about the utility of classifying culling reasons according to the voluntary nature of the culling decision. We concluded that Michigan producers are not using parents' genetics in heifer-culling decisions. PMID- 10673057 TI - Herd-based monitoring for tuberculosis in extensive swedish deer herds by culling and meat inspection rather than by intradermal tuberculin testing. AB - The effect of random slaughter and meat inspection as a tool to detect or eradicate tuberculosis in large, extensive deer herds in Sweden was evaluated. A computer spreadsheet model based on the Reed-Frost method was developed. Numbers of new infections and of infected deer slaughtered as well as probability of detecting tuberculosis or slaughtering all infected deer in a herd, were simulated. The model predicted that, given a 20% annual slaughter and that disease was introduced with one infected deer, the infection would be detected or eliminated in most herds (90%) after 15 years. PMID- 10673058 TI - Lung-tissue extract as an alternative to serum for surveillance for brucellosis in chamois. AB - Serological surveys are the most-used way to study diseases in free-ranging wild animals. However, the difficulty in obtaining a sufficient number of suitable serum samples is a major problem. To resolve this problem, we investigated the possibility of using lung-tissue extract in place of blood serum for searching for antibodies against Brucella abortus. Antibodies titres against B. abortus was tested from blood serum and lung-tissue extract from 112 chamois and 99 cattle. Although in complement-fixation-test, lung-tissue extract titres usually were one to three-fold lower than serum titres, there was a good agreement between serum and lung-tissue extract positivity both in chamois and in cattle. The lung-tissue extract appears a suitable resource in monitoring brucellosis in chamois. PMID- 10673059 TI - Influence of new infection with bovine virus diarrhoea virus on udder health in Norwegian dairy cows. AB - A retrospective longitudinal study was conducted to examine whether the exposure of dairy herds to bovine virus diarrhoea virus (BVDV) affected udder health. All Norwegian dairy herds that had experienced a marked increase in the BVDV antibody titres in bulk milk (from a level corresponding to an optical density (OD) <0.25 to >0.55, as determined by an indirect enzyme-linked immunosorbent assay) between two nation-wide herd screening examinations carried out late in 1992 and 1993, respectively, were considered to have been exposed to BVDV during the period between the examinations. The reference group included all dairy herds in which the bulk milk was BVDV antibody-negative or had only very low titres of BVDV antibodies (OD <0.25) each year from 1992 to 1995. The annual incidence rate of clinical mastitis, the bulk-milk somatic-cell count, and the annual rate of culling because of mastitis in the herds, were compared in the year of BVDV exposure (1993) as well as in a period prior to exposure (from 1988 to 1992) and two years following the year of exposure. In herds exposed to BVDV, there was a 7% increase in the incidence rate of clinical mastitis in the year of exposure, as compared with the nonexposed herds. No significant changes attributable to BVDV exposure were observed in the bulk-milk somatic-cell count or in the rate of culling because of mastitis. PMID- 10673060 TI - Isolation and characterization of cholic acid 7alpha-dehydroxylating fecal bacteria from cholesterol gallstone patients. AB - BACKGROUND/AIMS: The development of cholesterol gallstones, in some patients, has been associated with increased proportions of deoxycholic acid in the bile acid pool. Deoxycholic acid is a microbial product of cholic acid 7alpha dehydroxylation in the intestines. The levels and activities of bile acid 7alpha dehydroxylating bacteria have been reported to be increased in gallstone patients. The aim of the current study was to isolate 7alpha-dehydroxylating bacteria from gallstone patients and determine if these individuals are colonized by similar bacterial species. METHODS: The levels of 7alpha-dehydroxylating bacteria in fecal samples were determined by fecal dilutions in 24 gallstone patients and 10 controls. 7alpha-Dehydroxylating bacteria were isolated by a non selective streak plate technique and 7alpha-dehydroxylation activity was determined by measuring the conversion of [14C]-cholic acid to [14C]-deoxycholic acid using thin-layer chromatography. RESULTS: Gallstone patients had >42-fold (p<0.01) higher levels of 7alpha-dehydroxylating bacteria than patients who had not developed gallstones. Eighteen strains of 7alpha-dehydroxylating bacteria were isolated from eight gallstone patients. Attempts to isolate 7alpha dehydroxylating bacteria from ten control patients were unsuccessful using identical isolation techniques. Surprisingly, all strains of bacteria isolated from gallstone patients appear to belong to the genus Clostridium. CONCLUSION: Gallstone patients have higher levels of 7alpha-dehydroxylating fecal bacteria and appear to harbor only members of the genus Clostridium with this activity. PMID- 10673061 TI - Development of gap junctional channels and intercellular communication in rat liver during ontogenesis. AB - BACKGROUND/AIMS: We investigated the expression of connexin (Cx) 32 and 26 subunit proteins of the gap junction (GJ) in the rat liver during ontogenesis to clarify their roles in control of growth and differentiation, and observed their channels in association with development of gap junctional intercellular communication (GJIC). METHODS: The expression of Cx32 and 26 in prenatal and postnatal livers was examined by Western blot and immunofluorescence. GJ channels were investigated not only by double immunofluorescence study but also by immunogold electron microscopy. The spread of lucifer yellow 5 min after its microinjection was examined in the cultured liver tissues. RESULTS: 1) Western blot showed the expression of both Cx from the late stage of gestation and their peak a week after birth. 2) Cx32- or 26-positive plaques were scattered on hepatocytes of the fetal liver and some of them were colocalized; both were increased just after birth. On day 7 after birth, Cx32-positive plaques were present on all hepatocytes within a lobule, and Cx26-positive plaques were distributed in the periportal area. 3) Double-immunogold electron microscopy just after birth showed that most GJ channels were homotypic type of Cx32 or 26, and that few were heterotypic. On day 7 after birth, most channels had the homotypic type of type of Cx32 in the middle and pericentral areas, and there was a heterotypic type of Cx32 and 26 in the periportal area. 4) The dye transfer of lucifer yellow showed a wider spread in the liver tissues on day 7 after birth than on day 1. CONCLUSION: Increased GJ formation and compatibility or incompatibility of GJ channels are closely associated with development of GJIC, and GJIC may develop at cytodifferentiation during ontogenesis. PMID- 10673062 TI - Predictors of morbidity and mortality after the first episode of upper gastrointestinal bleeding in liver cirrhosis. AB - BACKGROUND/AIMS: Upper gastrointestinal (GI) bleeding is one of the most frequent causes of morbidity and mortality in the course of liver cirrhosis. The aim of this study was to determine the independent predictors of morbidity, mortality, and survival after the first episode of GI bleeding in patients with liver cirrhosis. METHODS: In a retrospective study of 403 cirrhotic patients who were admitted in the period January 1982 to December 1994 because of a first episode of GI hemorrhage, epidemiological factors, bleeding-related variables and cirrhosis-related variables that may be associated with hepatic and extrahepatic complications, mortality at 48 h and 6 weeks, and survival up to 30 June 1996 were assessed. RESULTS: Forty-five percent of patients developed hepatic and/or extrahepatic complications, with a mortality rate of 7.4% at 48 h and 24% at 6 weeks. Renal failure, rebleeding, hepatocellular carcinoma, and hepatic encephalopathy were independent predictors of mortality. The Kaplan-Meier method showed a median survival of 30.9+/-4.5 months (95% confidence interval 22 to 39.7 months). The cumulative percentage of survivors was 60.2% at 1 year, 33.6% at 5 years, and 14% at 10 years. In a Cox's multiple regression analysis, age, hepatic encephalopathy, hepatocellular carcinoma, Child-Pugh grade, and renal failure were independently associated with long-term survival. CONCLUSIONS: The first episode of GI bleeding in patients with liver cirrhosis is associated with high morbidity and mortality. Renal failure, rebleeding, hepatocellular carcinoma, and hepatic encephalopathy were independent risk factors for early death. PMID- 10673063 TI - Enhanced monocyte activation and hepatotoxicity in response to endotoxin in portal hypertension. AB - BACKGROUND/AIMS: Septic shock is a systemic response to infection, and it causes a high mortality rate in cirrhotic patients. The mechanisms responsible for this susceptibility in cirrhosis are poorly understood. The aim of this study was to investigate whether monocyte activation and hepatic function are altered in portal hypertension after endotoxin administration. METHODS: Portal-hypertensive and sham-operated rats were used. Plasma levels of tumor necrosis factor-alpha after lipopolysaccharide stimulation (both in vivo and in vitro) were measured by ELISA. CD11b/CD18 integrin expression on leukocyte membrane was measured by flow cytometry. Plasma transaminase activities were also determined. RESULTS: The levels of tumor necrosis factor-alpha in plasma and the expression of CD11b/CD18 on leukocytes in portal-hypertensive rats was similar to that in sham-operated rats. Injection of 150 microg/kg of lipopolysaccharide produced a 9-fold increase in plasma levels of tumor necrosis factor-alpha in portal-hypertensive compared with sham-operated rats, together with a significant up-regulation of CD11b/CD18 expression on monocytes and an elevation in plasma transaminase activity. Blood leukocytes incubated in vitro with lipopolysaccharide (0.5 microg/ml) induced a hypersecretion of tumor necrosis factor-alpha in portal-hypertensive rats, as compared to sham-operated rats. CONCLUSIONS: This study shows that monocytes from portal-hypertensive rats have an enhanced response to endotoxin, leading to hepatotoxicity. PMID- 10673064 TI - Pressor and renal effects of cross-linked hemoglobin in anesthetized cirrhotic rats. AB - BACKGROUND/AIMS: Cross-linked hemoglobin (XL-Hb), a hemoglobin-based oxygen carrier, is currently under investigation as a blood substitute. In the present study we have evaluated its pressor and renal effects in a rat model of liver cirrhosis by bile duct ligation. METHODS: Experiments were performed 3 weeks after surgery in anesthetized rats In the first protocol, the ability of XL-Hb to recover blood pressure after a hypotensive hemorrhage (0.5 ml/min, 10 min) was analyzed. In the second protocol, the pressor and renal effects produced by the administration of XL-Hb were evaluated during a period of 3 h. RESULTS: After a hypotensive hemorrhage (0.5 ml/min, 10 min), resuscitation with XL-Hb resulted in greater and faster recovery of blood pressure than with the administration of blood. In non-hemorrhaged rats, administration of XL-Hb (5% of blood volume) reversibly increased blood pressure in bile duct ligation and in control rats, but this effect was of longer duration in the control animals. XL-Hb also induced brisk increases in water and sodium excretion in both groups of animals, but the response of the control animals was more intense and sustained than that of the bile duct ligation rats. Glomerular filtration rate and renal blood flow showed slight decreases, but they were well maintained around the baseline levels. All the parameters studied were normalized 3 h later. In additional experiments, the effect of a bolus of L-NAME (10 mg/kg), an inhibitor of nitric oxide synthase, 1 h after the administration of XL-Hb was partially reduced, suggesting that the effect of XL-Hb may be secondary to the disappearance of circulating nitric oxide. CONCLUSIONS: XL-Hb seems to be effective as a resuscitative solution in case of hemorrhage in cirrhotic rats Moreover, this blood substitute only moderately and reversibly elevates blood pressure and does not adversely affects renal function. PMID- 10673065 TI - Aquaretic effects of niravoline, a kappa-opioid agonist, in patients with cirrhosis. AB - BACKGROUND/AIMS: In patients with cirrhosis, decreased renal water excretion is a common complication. Niravoline (RU51599), a kappa-opioid receptor agonist, has been shown to induce an aquaretic response. The aim of this study was to evaluate the aquaretic effect and tolerance of niravoline in patients with cirrhosis. METHODS: Biochemical tests and hemodynamic values were determined before and 1, 2, 3 and 24 h after niravoline administration at doses ranging from 0.5 to 2 mg iv in 18 patients with cirrhosis. RESULTS: Diuresis significantly increased in the first hour from 64+/-9 to 146+/-31 ml/h, and returned to basal values after 3 h. Free water clearance also significantly increased, reaching the positive range at 1 h. Plasma osmolality significantly decreased at 2 h (from 290+/-4 to 286+/-4 mOsm/kg). Plasma sodium concentrations increased significantly at 3 h (from 133+/ 1 to 134+/-1 mEq/l). Heart rate and arterial pressure did not change. The highest doses (1.5 mg or 2 mg) induced personality disorders and mild confusion within 2 h. These effects reversed completely within 8 h. CONCLUSION: This study shows that niravoline administration induces an aquaretic response and is well tolerated, at moderate doses, in patients with cirrhosis. Thus, moderate doses of niravoline may be useful for treating patients with cirrhosis and water retention. PMID- 10673066 TI - Mixed endothelin receptor antagonist, SB209670, decreases portal pressure in biliary cirrhotic rats in vivo by reducing portal venous system resistance. AB - BACKGROUND/AIMS: This study aimed to evaluate the hemodynamic effects of endothelin-1 or mixed endothelin receptor antagonist, SB209670 in cirrhotic rats, and to elucidate the role of endothelin in cirrhotic portal hypertension. METHODS: Secondary biliary cirrhosis was induced by bile duct ligation. Hemodynamics were studied using the radioactive microsphere technique. RESULTS: Plasma and hepatic endothelin levels in cirrhotic rats were significantly higher than those in normal rats (plasma, 9.0+/-1.3 vs. 2.6+/-0.5 pg/ml, p<0.001; liver, 74.8+/-13.3 vs. 12.6+/-2.5 pg/g wet tissue, p<0.001). Intraportal administration of endothelin-1 (3 nmol/kg) progressively raised portal pressure without an initial transient reduction, which was observed in systemic arterial pressure, in both cirrhotic and normal rats. SB209670 (5.4 micromol/kg) reduced portal pressure in cirrhotic rats (-19+/-5%, p<0.01) without modifying systemic arterial pressure and renal blood flow, but not in normal rats. This reduction was associated with reduced portal venous system resistance (vehicle, 2.5+/-0.2 vs. SB209670, 1.7+/-0.1 mmHg x min x 100 g bw/ml, p<0.01), but not with change in portal venous inflow and collateral blood flow. CONCLUSIONS: Mixed endothelin antagonist, SB209670, decreased portal pressure by reducing portal venous system resistance without modifying systemic arterial pressure and renal blood flow in cirrhotic rats. This result, together with the findings that plasma and hepatic endothelin levels were elevated in cirrhotic rats and that exogenous endothelin-1 increased portal pressure, provides further support for a role of endothelin in portal hypertension and suggests a potential use of mixed endothelin antagonist in the pharmacological treatment of portal hypertension. PMID- 10673067 TI - Gene expression of TNF-receptors in peripheral blood mononuclear cells of patients with alcoholic cirrhosis. AB - BACKGROUND/AIMS: Elevated concentrations of tumor necrosis factor receptors have been detected in alcoholic cirrhosis, but it remains unknown whether or not peripheral blood mononuclear cells are a source of tumor necrosis factor receptors and reflect the clinical disease activity of patients with advanced alcoholic liver disease. METHODS: Twenty-two abstinent patients in different stages of alcohol-induced cirrhosis according to the criteria of the Child-Pugh classification (Child-Pugh stage A: 4, Child-Pugh stage B: 10, Child-Pugh stage C: 8) were compared with four healthy individuals. Semi-quantitative reverse transcriptase-polymerase chain reaction was used for the measurement of the expression of tumor necrosis factor-alpha, soluble tumor necrosis factor receptors-p55, -p75, interleukin-10 and inducible nitric oxide synthase in unstimulated peripheral blood mononuclear cells. RESULTS: Unstimulated peripheral blood mononuclear cells of patients with alcoholic cirrhosis demonstrate a stage dependent enhanced RNA expression of tumor necrosis factor-alpha (healthy controls 0/4, Child-Pugh stage A 2/4, stage B 10/10, stage C 8/8; p<0.01). The mRNA expression of TNF-receptors-p55/-p75 is significantly higher in patients with severe alcoholic cirrhosis (Child-Pugh stage B or C patients) than healthy controls (p<0.05), while peripheral blood mononuclear cells from patients with Child-Pugh stage A show a similiar pattern of gene expression to healthy controls. No significant up-regulation of interleukin-10 was found. Inducible nitric oxide synthase was detectable in Child-Pugh stage C (p<0.05). CONCLUSIONS: Unstimulated peripheral blood mononuclear cells of patients with severe alcoholic cirrhosis (Child-Pugh stage B and C) demonstrate a systemic leukocyte activation and gene expression of tumor necrosis factor-alpha and tumor necrosis factor receptors-p55/-p75, which is correlated with the activity of the disease. Our data confirm previous studies that reported a correlation between plasma levels of pro-inflammatory cytokines and the severity of alcoholic cirrhosis. The role of interleukin-10 and inducible nitric oxide synthase in the pathogenesis of alcoholic cirrhosis remains to be fully elucidated. PMID- 10673068 TI - Protection by glycine against hypoxic injury of rat hepatocytes: inhibition of ion fluxes through nonspecific leaks. AB - BACKGROUND/AIMS: Glycine has long been shown to exert strong protective effects against hypoxic injury of hepatocytes. Recently, it was suggested that glycine exerts this protection via inhibition of ligand-gated chloride channels, thereby secondarily inhibiting sodium influx. The purpose of this study was to examine this suggestion. METHODS: Cultured rat hepatocytes were incubated under normoxic and hypoxic conditions. Loss of viability was determined by release of lactate dehydrogenase. Cytosolic ion concentrations were measured using digital fluorescence microscopy. RESULTS: Glycine prevented the hypoxic increase in cytosolic sodium and strongly protected against hypoxic injury. The amino acid was not only protective in Krebs-Henseleit buffer but also in a chloride-free modification thereof and offered additional protection in a sodium-free medium (which already yielded substantial protection in its own right). Glycine also prevented the hypoxic release of the anionic fluorescent dye Newport Green and appeared to prevent the hypoxic entrance of the "nonphysiological" cations cobalt and nickel. CONCLUSION: The results strongly argue against inhibition of ligand gated chloride channels as being responsible for the potent protective effect of glycine against hypoxic injury of hepatocytes. Instead, they suggest that glycine prevents the formation of nonspecific leaks for small ions including sodium, thereby providing protection. PMID- 10673069 TI - Lovastatin decreases mortality and improves liver functions in fulminant hepatic failure from 90% partial hepatectomy in rats. AB - BACKGROUND/AIMS: Liver insufficiency occurs when the liver cannot perform critical functions such as ammonia metabolism, gluconeogenesis, or production of coagulation factors The hypothesis of this study was that decreased function of existing hepatocytes may contribute to hepatic failure, and that the function of these cells might be increased pharmacologically. Lovastatin is a 3-hydroxy-3 methylglutaryl CoA reductase inhibitor that inhibits cholesterol biosynthesis and affects the activity of some signal transduction pathways and liver transcription factors. Changes in hepatic transcription factors during liver regeneration might result in decreased liver functions, and lovastatin might prevent these changes METHODS: Rats received 90% partial hepatectomy (90% PH), and either lovastatin or vehicle alone daily. Survival and liver functions were assessed. RESULTS: Lovastatin increased survival to 58% (vs. 6% in controls that received 90% PH without drug), decreased the peak ammonia level to 427 microM (vs. 846 microM in controls), increased the nadir of glucose to 88 mg/dl (vs. 57 mg/dl in controls), decreased the peak prothrombin time to 23 s (vs 29 s in controls), and decreased the peak activated partial thromboplastin time to 29 s (vs. 39 s in controls). The full survival and metabolic benefits were observed when lovastatin was started at 30 min after 90% PH, but lovastatin was less efficacious when started at later times. CONCLUSIONS: Lovastatin increases the function of existing hepatocytes and might be used to improve liver function after extensive hepatic resection. PMID- 10673070 TI - Protein expression of CD44 (standard and variant isoforms) in hepatocellular carcinoma: relationships with tumor grade, clinicopathologic parameters, p53 expression, and patient survival. AB - BACKGROUND/AIMS: Members of the CD44 family are transmembrane glycoproteins which act mainly as receptors for hyaluronan. We have examined the expression of CD44s and several CD44v and the relationship between these and hepatocellular carcinoma (HCC) grade, clinicopathological parameters, p53 expression, and patient survival in HCC. METHODS: Formalin-fixed, paraffin-embedded tissue sections from 107 surgically resected HCC were examined immunohistochemically using a semi quantitative scoring system to detect the expression of different forms of CD44. RESULTS: The number of CD44s-positive cases was 36 (34%), CD44v5 52 (49%), CD44v6 29 (27%), CD44v7-8 41 (38%), and CD44v10 26 (24%). Expression of these molecules correlated with high histological grade, being the highest in poorly differentiated HCC. High CD44v6 expression significantly correlated with the presence of vascular invasion and p53 overexpression. Kaplan-Meier examination of patient survival revealed that HCC patients with positivity of each of these five molecules had a reduced survival rate, and that HCC patients positive for all the five CD44 molecules had worse survival than HCC patients positive for four or less of these CD44 molecules. In multivariate survival analysis, CD44s positivity was an independent factor. However, positivity for one or more CD44 isoforms was the most useful independent factor for overall survival. CONCLUSION: These results suggest that up-regulation of CD44 isoforms is associated with poorly differentiated HCC and shortened survival. PMID- 10673071 TI - Increased GABAergic activity inhibits alpha-fetoprotein mRNA expression and the proliferative activity of the HepG2 human hepatocellular carcinoma cell line. AB - BACKGROUND/AIMS: Gamma aminobutyric acid (GABA) is a potent inhibitory neurotransmitter with growth regulatory properties. Recent data indicate that increased GABAergic activity inhibits hepatocyte proliferation in regenerating livers. In the present study, we aimed to investigate whether GABA inhibits the growth of malignant hepatocytes. METHODS: Increasing concentrations of muscimol (0.05-50 microM), a specific GABA(A) receptor agonist, were added to HepG2 human hepatocellular carcinoma cells and alpha-fetoprotein (AFP) and albumin mRNA expression were determined for varying periods of time (maximum 24 h) thereafter. Cell proliferation was also documented after 48 h of exposure to muscimol. RESULTS: Muscimol significantly (p<0.0001) decreased AFP mRNA expression (maximum decrease: 65% below baseline values) without affecting albumin mRNA expression. However, the effect on AFP mRNA was transient (maximum duration: 3-6 h) and not associated with changes in cell proliferation. Because preliminary data indicate that GABA(A) receptor activity is markedly downregulated in malignant hepatocytes, transfection studies were performed wherein HepG2 cells were cotransfected with GABA(A) receptor beta2 and beta2 subunit genes in a pCDM8 expression vector or vector alone followed by re-exposure to either muscimol (5 betaM) or saline. In this series of experiments, in addition to AFP mRNA inhibition being as extensive and more prolonged (maximum duration: 6-12 h) in muscimol-treated, GABA(A) receptor-transfected cells, proliferative activity was also significantly inhibited when compared to saline-treated GABA(A) receptor transfected controls (p<0.01) and muscimol-treated cells transfected with vector alone (p<0.005). CONCLUSION: The results of this study indicate that increased GABAergic activity inhibits AFP mRNA expression and cell proliferation in this malignant hepatocyte cell line. PMID- 10673072 TI - A possible novel src-related tyrosine kinase in cancer cells of LEC rats that develop hepatocellular carcinoma. AB - BACKGROUNDS/AIMS: Src-related protein tyrosine kinase is known to be related to cell transformation. In this study, we report a possible novel src-related tyrosine kinase of 100 kDa specifically expressed in the nuclei of hepatocytes and/or cancer cells in Long-Evans Cinnamon rats, one of the experimental models of hepatocellular carcinoma. METHODS: Src-related protein tyrosine kinase in hepatocytes of Long-Evans Cinnamon rats was analyzed by using immunohistochemistry and Western blot and in vitro tyrosine kinase assay using a specific antibody (src antibody) against a synthetic peptide corresponding to the conserved autophosphorylation site of src family tyrosine kinases. RESULTS: Src related protein was found to be expressed in the nuclei of hepatocytes and/or cancer cells in Long-Evans Cinnamon rat liver, exhibiting tyrosine kinase activity, and migrated to the position of 100 kDa. The protein quantity and activity of this 100-kDa src-related protein tyrosine kinase significantly increased with the progress of chronic hepatitis to hepatocellular carcinoma, especially in the tumorous portion of the liver. On the other hand, the 100-kDa src-protein tyrosine kinase was not observed in the nuclei of hepatocytes and/or cancer cells in normal age-matched control Wistar rats. CONCLUSIONS: Since the src-family tyrosine kinases have been observed at a molecular weight of 55 to 62 kDa and located in the hepatocellular membrane and/or cytoplasm, the 100-kDa src related protein tyrosine kinase observed in the present study may be novel, and closely related to the pre-cancerous and cancerous process in Long-Evans Cinnamon rat liver. PMID- 10673073 TI - Characteristics of Epstein-Barr virus primary infection in pediatric liver transplant recipients. AB - BACKGROUND/AIM: Pediatric liver transplant recipients are at high risk of Epstein Barr virus infection. However the incidence of clinical symptoms and the graft function at the time of acute infection remains poorly documented. The aim of this study was to monitor the clinical and biochemical events associated with primary Epstein-Barr virus infection. METHODS: Clinical and biological patterns associated with Epstein-Barr virus infection were prospectively searched in 38 liver transplanted children. Polymerase chain reaction and anti-Epstein-Barr virus IgM antibodies were used at regular intervals to detect the timing of primary infection. RESULTS: Five children (13%) had pretransplant immunity, 26 (68.5%) developed primary Epstein-Barr virus infection 15 to 90 days after transplantation and seven (18.5%) remained Epstein-Barr virus negative. The four patients with clinical symptoms at the time of infection subsequently developed post-transplant lymphoproliferative disease. A single post-transplant lymphoproliferative disease occurred in non-symptomatic patients (overall incidence 13%). No mortality was associated with post-transplant lymphoproliferative disease. Two asymptomatic patients had abnormal liver function tests possibly related to primary Epstein-Barr virus infection. CONCLUSION: Epstein-Barr virus primary infection occurs in 80% of seronegative patients within 3 months after OLT. Clinical symptoms are rare and closely associated with post-transplant lymphoproliferative disease. Outside post transplant lymphoproliferative disease, the consequences of infection are marginal. PMID- 10673074 TI - Cold preservation of fatty liver grafts: prevention of functional and ultrastructural impairments by venous oxygen persufflation. AB - BACKGROUND/AIMS: The incidence of steatosis in livers retrieved for organ transplantation is up to 30%. Due to the shortage of donor organs, many of these livers are accepted for clinical transplantation, although a high rate of graft dysfunction is associated with ischemic preservation of steatotic livers. The present study was intended to reduce the ischemia/reperfusion injury of steatotic grafts by the use of venous systemic oxygen persufflation during cold storage. METHODS: A histologically-documented mild to moderate steatosis was induced in livers of Wistar rats by fasting for 2 days and subsequent feeding of a fat-free diet enriched in carbohydrates. Fatty livers were retrieved and flushed via the portal vein with 60 ml of HTK. In group A, livers were then stored ischemically at 4 degrees C for 24 h. Livers of group B were additionally connected to a gaseous oxygen supply and persufflated with O2 via the venous vascular system during the cold storage period. Viability of the livers was then assessed upon isolated perfusion in vitro with oxygenated Krebs-Henseleit buffer. RESULTS: Venous systemic oxygen sufflation resulted in a relevant and significant reduction of parenchymal (ALT: 132+/-90 vs 434+/-172 U/l; p<0.01) and mitochondrial (GLDH: 116+/-57 vs 633+/-241 U/l; p<0.001) enzyme release during reperfusion. Moreover, Kupffer cell activation, as evaluated from acid phosphatase activity in the perfusate, was reduced to about 1/3 (4.0+/-1.3 vs 11.9+/-5.3 U/l; p<0.01). Electron microscopic analysis revealed that the liver mitochondria and sinusoidal endothelial lining were better preserved after oxygen persufflation, which was in line with the data on enzyme release and the increased portal perfusion pressure in the untreated group, while normal values were found after venous systemic oxygen sufflation. CONCLUSION: Venous oxygen persufflation may thus represent a useful tool for the safe and improved preservation of ischemia-sensitive steatotic livers. PMID- 10673075 TI - Endotoxin-induced aggravation of preservation-reperfusion injury of rat liver and its modulation. AB - BACKGROUND/AIMS: In clinical transplantation, exposure of donors to gut-derived endotoxin occurs frequently and may adversely affect liver transplantation therapy. The aim of this study was to investigate: 1) whether brief exposure of rats to endotoxin before liver procurement aggravates the early phase of reperfusion injury of hepatic explants; and if so 2) whether Kupffer cell activation is a contributing factor to liver injury; and 3) whether heparin and pentoxifylline could minimize this effect. METHODS: Male Wistar rats were injected with 0.2-4.0 mg/kg of Escherichia coli lipopolysaccharide 2 h prior to liver harvest. After preservation in University of Wisconsin cold-storage solution, the livers were reperfused using a blood-free perfusion model. To inactivate Kupffer cells, some rats were pretreated with gadolinium chloride or liposome-encapsulated dichloromethylene-diphosphonate before lipopolysaccharide administration. The other rats received lipopolysaccharide with heparin or pentoxifylline. RESULTS: In a dose-independent fashion, lipopolysaccharide impaired portal flow during graft reperfusion. In a dose-dependent way, lipopolysaccharide increased lactate dehydrogenase release into the perfusate and decreased bile flow and bromosulfophthalein excretion. Gadolinium chloride, liposomal dichloromethylene-diphosphonate, heparin, and pentoxifylline reduced lactate dehydrogenase release by 34%, 43%, 59%, and 64%, respectively, and improved functional parameters of the liver. A 52-fold increased neutrophil infiltration in the liver sinusoids after lipopolysaccharide exposure was not affected significantly by the drugs studied; however, heparin reduced markedly neutrophil activation. CONCLUSIONS: The results of this investigation provide direct evidence that aggravation of preservation-reperfusion injury of rat liver by endotoxin is mediated by Kupffer cell-dependent mechanism(s) and it can be minimized by heparin and pentoxifylline. PMID- 10673076 TI - Expansion of peripheral blood CD5+ B cells is associated with mild disease in chronic hepatitis C virus infection. AB - BACKGROUND/AIMS: Hepatitis C virus (HCV) infection is associated with the development of chronic liver disease and extra-hepatic manifestations, which include autoantibody production, immune-mediated diseases such as cryoglobulinaemia and B-cell lymphoproliferation. Recent identification of intra hepatic clonal B cells capable of rheumatoid factor production, selective infection of B cells over T cells and of an HCV receptor on B lymphocytes strongly supports a central role for these cells in the immune response to HCV infection. In particular, CD5+ B cells which are capable of producing natural antibodies with autoreactive specificities are likely to be important in the development of HCV-associated autoimmunity and lymphoproliferation. METHODS: We have investigated the presence of CD5+ B cells in a unique cohort of HCV-infected women who were infected with a single inoculum of HCV genotype 1b following immunisation with contaminated anti-D immunoglobulin in 1977. RESULTS: CD5+ B cells are significantly increased in chronic HCV infection (37.66+/-1.92%) as compared with those with resolved infection (25.33+/-1.90%). High levels of CD5+ B cells were associated with the production of rheumatoid factor. The number of peripheral blood CD5+ B cells correlated negatively with histological activity index. CONCLUSIONS: The expansion of this B cell population in patients with active HCV infection may give rise to immune-mediated sequelae associated with HCV infection. This expanded population of CD5+ B cells may protect against the development of progressive liver disease. PMID- 10673077 TI - Post-transplant quasispecies pattern remains stable over time in patients with recurrent cholestatic hepatitis due to hepatitis C virus. AB - BACKGROUND/AIMS: Several studies have shown that cholestatic recurrent hepatitis is associated with very high HCV RNA loads in liver transplant recipients. The aim of this study was to investigate whether a correlation exists between cholestatic hepatitis post-transplant and the population of viral quasispecies. METHODS: One hundred and nine serial sera samples were tested from 15 recurrent HCV patients. Four of these patients showed severe cholestatic recurrent hepatitis, 11 patients demonstrated non-cholestatic recurrent hepatitis post transplant. Quasispecies were detected by RT-PCR amplification of the HVR1 followed by single-stranded conformational polymorphism analysis. RESULTS: Forty one samples from four cholestatic patients were tested. All four patients showed very stable quasispecies patterns post-transplant. One cholestatic patient also showed a stable quasispecies band pattern following retransplantation, again associated with severe cholestatic hepatitis. Sixty-eight samples were tested from the 11 non-cholestatic patients. In contrast, these patients showed significantly more quasispecies bands than the cholestatic patients. The noncholestatic patients also displayed fluctuating band patterns post-transplant. Serial samples were tested after retransplantation in one non-cholestatic patient, with a fluctuating pattern again seen. There was a negative correlation between the HCV RNA load in serum and the number of quasispecies bands. CONCLUSIONS: Stable hepatitis C viral quasispecies associated with persistently high viral load in post-transplant cholestatic hepatitis suggest that viral escape from immune pressures may play a role in the pathogenesis of this condition. PMID- 10673079 TI - Diagnosis, treatment and prophylaxis of spontaneous bacterial peritonitis: a consensus document. International Ascites Club. PMID- 10673078 TI - Mitochondrial membrane perturbations in cholestasis. PMID- 10673080 TI - Demonstration of McCune-Albright mutations in the liver of children with high gammaGT progressive cholestasis. AB - Two patients presented with neonatal cholestasis and acholic stools as first manifestations of McCune-Albright syndrome. Both went through an extensive evaluation including an exploratory laparotomy with peroperative cholangiography which ruled out biliary atresia. One patient presented from the fourth month of life with the classical cafe-au-lait spots following Blaschko's lines, while less classical cafe-au-lait spots were seen in the second patient at the age of 4 years. Bone lesions were seen in one patient at the age of 2.5 years and in the other at the age of 4 years. Despite the severity of presentation, both patients cleared their jaundice within 6 months, but still had mild abnormalities of liver function tests. Both patients showed an activating mutation of codon 201 in the gene encoding the alpha-subunit of the G-protein that stimulates adenylcyclase in liver tissue, suggesting that this metabolic defect could be responsible for the cholestatic syndrome. Similar mutations have been found in other affected tissues in patients with the McCune-Albright syndrome. We propose that McCune-Albright syndrome be included in the list for differential diagnosis of neonatal cholestasis and chronic cholestasis of infancy, as a rare cause. PMID- 10673082 TI - Pericardial disease is often not recognised as a cause of chronic severe ascites. AB - BACKGROUND/AIMS: Severe chronic ascites remains a difficult diagnostic and therapeutic problem. Even in the current era, constrictive pericarditis is an underestimated and sometimes unrecognised cause. Moreover, missing the diagnosis deprives patients of remedial therapy. METHODS: Two cases of calcified constrictive pericarditis, complicated with cirrhosis and diagnosed in a late stage, are described. Due to insufficient clinical appreciation and lack of trust in echocardiography results, performed by cardiologists who were insufficiently familiar with the echocardiographic features of constrictive pericarditis, diagnosis was delayed in the two patients RESULTS: The diagnosis of constrictive pericarditis as a cause of ascites is based upon the clinical signs of right heart failure in a patient with normal systolic left and right ventricular function and a high, serumascitic albumin-content difference. Complementary workup with complete Doppler echocardiography study, right and left heart catheterisation and MRI or cine CT of the heart is necessary to confirm the diagnosis. CONCLUSION: Careful history taking and clinical examination remain the cornerstone of any diagnostic work-up, even in this era of technological refinement. PMID- 10673081 TI - Familial idiopathic adulthood ductopenia: a report of five cases in three generations. AB - BACKGROUND/AIMS: Idiopathic adulthood ductopenia is a cholestatic liver disease of unknown etiology. Although most cases are sporadic, familial cases do occur. METHODS: We describe a series of adult-onset bile duct depletion involving five members of an extended family spanning three generations. The proband, a 49-year old man, presented in 1989 with asymptomatic elevation of liver enzyme tests. Investigations for chronic liver disease, including endoscopic retrograde cholangiopancreatography, were negative. Findings on liver biopsy progressed from normal in 1989 to striking loss of interlobular bile ducts in 1992. Ursodeoxycholic acid has resulted in improvement of liver enzyme tests. The proband's brother required a liver transplant at age 35 for cryptogenic cirrhosis. The proband's sister, age 42, has had intermittent jaundice and elevation of liver enzyme tests since 1971. Her liver biopsy findings progressed from normal in 1975, to striking bile duct damage by 1997. The proband's 21-year old son has elevated liver enzyme tests and a liver biopsy consistent with idiopathic adulthood ductopenia. The proband's father had a liver biopsy at age 70 for investigation of a liver mass. It revealed extensive fibrosis and striking bile duct destruction. RESULTS/CONCLUSIONS: This is the largest series of familial idiopathic adulthood ductopenia reported, and the first with multiple generations described. Genetics appears to play a role in some cases of adulthood ductopenia. Ursodeoxycholic acid may be beneficial in the treatment of this condition. PMID- 10673083 TI - Liver failure due to herpes simplex virus. PMID- 10673084 TI - Two different doses and duration schedules of somatostatin -14 in the treatment of patients with bleeding oesophageal varices: a non-randomised controlled study. PMID- 10673085 TI - Interferon therapy in patients with chronic hepatitis C and schistosomiasis. PMID- 10673086 TI - Fatal hepatitis associated with nimesulide. PMID- 10673087 TI - Solution-phase combinatorial synthesis and evaluation of piperazine-2,5-dione derivatives. AB - An efficient one-pot synthesis of a 61-membered combinatorial chemistry library of piperazine-2,5-diones was accomplished. Results of combinatorial synthesis, purification, analysis, and biological evaluation are described. PMID- 10673088 TI - Synthesis and antibacterial activity of new carbapenems containing isoxazole moiety. AB - The synthesis and biological activity of a series of new 1beta-methylcarbapenems 1a-g containing 5'-isoxazolopyrrolidin-3'-ylthio derivatives as C-2 side chain are described. Most compounds exhibited potent and well-balanced antibacterial activity as well as high stability to DHP-I comparable to that of meropenem. 1e and 1c showed the best combination of antibacterial activity and stability to DHP I, respectively. PMID- 10673089 TI - Synthesis and biological activities of hapalosin derivatives with modification at the C12 position. AB - Among the hapalosin derivatives synthesized, the compounds carrying methyl (5a), methylthioethyl (5d) and phenylmethyl (5e) groups at the C12 position possess only the cis-peptide structure, in contrast to the cases of 5b and 5c. In addition to their conformational stability, the biological activities of the compounds were determined in relation of the P-glycoprotein-mediated MDR reversing activity and induction of apoptosis. PMID- 10673090 TI - Epicubenol synthase. Origin of the oxygen atom of a bacterial sesquiterpene alcohol. AB - Incubation of epicubenol synthase with farnesyl pyrophosphate in the presence of 11.1 atom% H2(18)O gave epicubenol (2) in which the hydroxyl oxygen atom was shown to be derived exclusively from water, as established by GC-selected ion monitoring MS of the derived TMS-epicubenol derivative (15). PMID- 10673091 TI - Structure-activity relationships of trans-3,5-disubstituted pyrrolidinylthio 1beta-methylcarbapenems. Part 1: J-111,347 and related compounds. AB - 1Beta-methylcarbapenems having various 3,5-disubstituted pyrrolidinylthio-side chains at C-2 were designed and synthesized. Evaluation of their antibacterial activities indicated that J-111,347 (1a) is the first example of an extremely broad spectrum antibiotic showing activity against methicillin-resistant Staphylococcus aureus (MRSA) as well as Pseudomonas aeruginosa. PMID- 10673092 TI - Structure-activity relationships of trans-3,5-disubstituted pyrrolidinylthio 1beta-methylcarbapenems. Part 2: J-111,225, J-114,870, J-114,871 and related compounds. AB - Through further derivatization of J-111,347 (1a), a trans-3,5-disubstituted pyrrolidinylthio-1beta-methylcarbapenem, undesired epileptogenicity in a rat intracerebroventricular assay (200 microg/rat) could be eliminated to afford J 111,225 (2a), J-114,870 (3a) and J-114,871 (3b) which preserved comparable broad antimicrobial activity. PMID- 10673093 TI - Structure-activity relationship study of asiatic acid derivatives against beta amyloid (A beta)-induced neurotoxicity. AB - 8 Semi-synthetic derivatives of asiatic acid were prepared and their protective effect against A beta-induced neurotoxicity was evaluated. Among them, asiatic acid (2), and 4, 16 showed 97, 92 and 87% of protective effect, respectively. PMID- 10673094 TI - Photosensitizers related to purpurin-18-N-alkylimides: a comparative in vivo tumoricidal ability of ester versus amide functionalities. AB - For a comparative study, 3-(alkyloxyethyl)-3-devinylpurpurin-18-N-hexylimides with ester and amide functionalities were investigated for tumor selectivity and in vivo photosensitizing efficacy. Compared to amide analogues, the related photosensitizers with ester functionalities were found to be more effective. Among these compounds the 3-devinyl-(3-hexyloxyethyl)-purpurin-18-N-hexylimide as methyl ester 12 showed excellent tumor uptake (tumor versus muscle ratio: 8:1), and produced 100% tumor cure on day 30 at a dose of 1.0 micromol/kg. The mice were treated with light (135 J/cm2, 705 nm) at 24 h post injection of the drug. PMID- 10673095 TI - First enantiospecific synthesis of a 3,4-dihydroxy-L-glutamic acid [(3S,4S) DHGA], a new mGluR1 agonist. AB - The first synthesis of one of the 4 possible stereoisomers of 3,4-dihydroxy-L glutamic acid ((3S,4S)-DHGA 3), a natural product of unknown configuration, is described. The synthesis is based on the Lewis acid catalyzed reaction of benzyl alcohol with a D-ribose-derived 2,3-aziridino-gamma-lactone 4-benzyl carboxylate (6). Preliminary pharmacological studies showed that (3S,4S)-3 is an agonist of metabotropic glutamate receptors of type 1 (mGluR1) and a weak antagonist of mGluR4 but has no discernible activity with respect to mGluR2. This activity profile can be rationalized by fitting extended conformations of (3S,4S)-3 in proposed models of each of these receptor subtypes. PMID- 10673096 TI - A new benzoangelicin with strong photobiological activity. AB - Benzoangelicins 4-6 were synthesized in good yields from 7-hydroxy-5-methoxy-4 methylcoumarin (1). In the absence of UVA radiation, compounds 5 and 6 were only weakly active against HL60 and HeLa tumour cells; in its presence, compound 6 was 10 times more active than the reference compound 8-methoxypsoralen. None of 4-6 exhibited cutaneous phototoxicity. PMID- 10673097 TI - 3'-Beta-ethynyl and 2'-deoxy-3'-beta-ethynyl adenosines: first 3'-beta-branched adenosines substrates of adenosine deaminase. AB - The 3'-C-branched-adenosine and 2'-deoxyadenosine analogues 1-7 were tested as substrate of adenosine deaminase. The 9-(3'-C-ethynyl-beta-D-ribo pentofuranosyl)adenine 1 and its 2'-deoxy analogue 7 were deaminated by the enzyme while the vinyl and ethyl derivatives 2 and 3 were not. The 9-(3'-C branched-beta-D-xylo-pentofuranosyl)adenines 4-6 were deaminated by the deaminase. PMID- 10673098 TI - Multi gram synthesis of UDP-N-acetylmuramic acid. AB - As part of an effort to discover novel antibacterial agents, a new and efficient synthesis was established in order to provide a large amount of UDP-N acetylmuramic acid (UDP-MurNAc). PMID- 10673099 TI - Synthesis and estrogenic activities of novel 7-thiosubstituted estratriene derivatives. AB - A diastereomerically pure series of 7alpha-thioestratrienes was prepared and evaluated for its affinity for both the human estrogen receptor alpha and the more recently discovered estrogen receptor beta. The functional estrogenic activities of the compounds were measured in a MCF-7 ERE-tk-luciferase assay. The activities and selectivities of the compounds were sensitive to the nature of the thioether side chain. PMID- 10673100 TI - Synthesis of ketomethylene amino pseudopeptide analogues via reductive amination of glyoxals derived from alpha-amino acids. AB - The reductive amination of an amino acid derived glyoxal, with the free amino group of a protected amino acid or oligopeptide fragment, has been developed as a simple and efficient method for the preparation of ketomethylene amino pseudo oligopeptide isosteres Aa psi(COCH2NH)Aa. Trichlorosilane-DMF is the reagent of choice for the reduction. PMID- 10673101 TI - The flavanolignan silybin and its hemisynthetic derivatives, a novel series of potential modulators of P-glycoprotein. AB - A new series of potential flavonoidic modulators of P-glycoprotein activity has been prepared. The flavanolignan silybin was first oxidised to dehydrosilybin and then C-alkylated with either prenyl or geranyl bromide. The resulting isoprenoid dehydrosilybins were shown to display high in vitro affinities for direct binding to P-glycoprotein, which ranged them among the best flavonoids ever tested. PMID- 10673102 TI - Coumarin inhibitors of gyrase B with N-propargyloxy-carbamate as an effective pyrrole bioisostere. AB - The synthesis and biological profile in vitro of a series of coumarin inhibitors of gyrase B bearing a N-propargyloxycarbamate at C-3' of noviose is presented. Replacement of the 5-methylpyrrole-2-carboxylate of coumarin drugs with an N propargyloxycarbamate bioisostere leads to analogues with improved antibacterial activity. Analysis of crystal structures of coumarin antibiotics with the 24 kDa N-terminal domain of the gyrase B protein provides a rational for the excellent inhibitory potency of C-3' N-alkoxycarbamates. PMID- 10673103 TI - Novel Dmt-Tic dipeptide analogues as selective delta-opioid receptor antagonists. AB - A series of Dmt-Tic analogues with substitution on the Tic aromatic ring has been synthesized and evaluated for opioid receptor affinity and activation. Incorporation of large hydrophobic groups at position 7 of Tic did not greatly alter the delta opioid receptor binding affinities of the dipeptides whereas substitution at position 6 substantially diminished their affinity. These modified Dmt-Tic peptides showed binding affinities as low as 2.5 nM with up to 500-fold selectivity for the delta versus mu opioid receptor and proved to be delta receptor antagonists. PMID- 10673104 TI - The use of power ultrasound coupled with magnetic separation for the solid phase synthesis of compound libraries. AB - Enhanced reaction rates are observed when power ultrasound is utilized as a substitute for mixing during solid phase organic chemical reactions on a paramagnetic support. Power ultrasound is also used to facilitate the washing of the paramagnetic support as it is magnetically separated from the reaction mixture. Selective examples from a library targeting the kappa-opioid receptor are presented. PMID- 10673105 TI - Synthesis and evaluation of furo[3,4-d]pyrimidinones as selective alpha1a adrenergic receptor antagonists. AB - Furo[3,4-d]pyrimidinones were found to be metabolites of dihydropyrimidinones such as 1a-b that are subtype-selective antagonists of the alpha1a-adrenergic receptor. A versatile synthesis that provides access to furo[3,4-d]pyrimidinones in high yield and in enantiomerically pure forms is described along with structure-activity relationships in the series. PMID- 10673106 TI - RGD mimetics containing a central hydantoin scaffold: alpkha(v)beta3 vs alpha(IIb)beta3 selectivity requirements. AB - The synthesis of a series of RGD mimetic alpha(v)beta3 antagonists containing a hydantoin scaffold is shown. The results demonstrate some of the structural requirements for the design of selective alpha(v)beta3 antagonists (vs alpha(IIb)beta3) in terms of the Arg-mimetic, the distance between N- and C terminus and the lipophilic side chain. PMID- 10673107 TI - 8-Aminocyclazocine analogues: synthesis and structure-activity relationships. AB - Opioid binding affinities were assessed for a series of cyclazocine analogues where the prototypic 8-OH substituent of cyclazocine was replaced by amino and substituted-amino groups. For mu and kappa opioid receptors, secondary amine derivatives having the (2R,6R,11R)-configuration had the highest affinity. Most targets were efficiently synthesized from the triflate of cyclazocine or its enantiomers using Pd-catalyzed amination procedures. PMID- 10673108 TI - Synthesis of dolichyl phosphate derivatives with fluorescent label at the omega end of the chain, new tools to study protein glycosylation. AB - Derivatives of dolichyl phosphate (Dol-P) with 2-aminopyridine or 1 aminonaphtalene fluorophore groups at the omega-end of the chain were synthesized. These products serve as substrates for recombinant yeast Dol-P mannose synthase. Fluorescence resonance energy transfer between a Trp residue of the enzyme and the 1-aminonaphtalene group of the Dol-P analogue was demonstrated. PMID- 10673109 TI - Novel 2,2-dioxide-4,4-disubstituted-1,3-H-2,1,3-benzothiadiazines as non nucleoside reverse transcriptase inhibitors. AB - Benzothiadiazine non-nucleoside reverse transcriptase inhibitors (NNRTIs) of HIV have been synthesized via a novel process to afford active inhibitors, with the most potent compound exhibiting an IC90 = 180 nM in a whole cell assay. The 2,2 dioxide-1H-2,1,3-benzothiadiazine ring system was constructed in one step from 2 amino-5-chlorobenzonitrile. PMID- 10673110 TI - Biomechanical analysis of the stance phase during barefoot and shod running. AB - This study investigated spatio-temporal variables, ground reaction forces and sagittal and frontal plane kinematics during the stance phase of nine trained subjects running barefoot and shod at three different velocities (3.5, 4.5, 5.5 m s(-1)). Differences between conditions were detected with the general linear method (factorial model). Barefoot running is characterized by a significantly larger external loading rate than the shod condition. The flatter foot placement at touchdown is prepared in free flight, implying an actively induced adaptation strategy. In the barefoot condition, plantar pressure measurements reveal a flatter foot placement to correlate with lower peak heel pressures. Therefore, it is assumed that runners adopt this different touchdown geometry in barefoot running in an attempt to limit the local pressure underneath the heel. A significantly higher leg stiffness during the stance phase was found for the barefoot condition. The sagittal kinematic adaptations between conditions were found in the same way for all subjects and at the three running velocities. However, large individual variations were observed between the runners for the rearfoot kinematics. PMID- 10673111 TI - Mechanical validation of whole bone composite tibia models. AB - Composite synthetic models of the human tibia have recently become commercially available as substitutes for cadaveric specimens. Their use is justified by the advantages they offer as a substitute for real tibias. The present investigation concentrated on an extensive experimental validation of the mechanical behaviour of the whole bone composite model, compared to human specimens for different loading conditions. The stiffness of the tibias was measured with a torsional load applied along the long axis, and with a bending load applied both in the latero-medial and in the antero-posterior direction. The bending stiffness of the composite tibias matched well with that of the cadaveric specimens. This was not true for the torsional stiffness. In fact, the composite tibias were much stiffer than the cadaveric specimens, possibly due to the structure of the reinforcement material. The inter-specimen variability for the composite tibias was much lower than that for the cadaveric specimens. Thus, it seems that the composite tibias are suitable to replace cadaveric specimens for certain types of test, whereas they might be unsuitable for others, depending on the loading regimen. PMID- 10673112 TI - Bone mineral density and bone structure parameters as predictors of bone strength: an analysis using computerized microtomography and gastrectomy-induced osteopenia in the rat. AB - In this study the relationships of bone mineral density (BMD) and bone structure parameters calculated from 2D microtomography images to bone strength were investigated. Femurs from 21 male Sprague Dawley rats were subjected to dual energy X-ray absorptiometry, computerized microtomography (CmicroT) and either three-point cantilever bending (femoral shaft) or two-point bending compression (femoral neck). Gastrectomy was performed on 12 animals and 9 were sham operated. From the tomograms bone structure analysis was performed using a software routine based on grey level run-length method. Correlations of BMD and bone structure parameters to mechanical parameters were investigated as were differences between the gastrectomized and the control samples. The reductions of BMD between the groups were 21 and 27% in the femoral neck and shaft, respectively. For the shaft, the correlations of BMD to all mechanical parameters were significant and BMD was a consistent predictor of bone strength for cortical bone. However, in the femoral neck where cancellous bone predominates, BMD was weakly correlated only to deflection. A significant correlation between trabecular thickness and neck bone strength was found. Hence, compared to trabecular thickness, BMD was of limited value in predicting bone strength in the femoral neck. PMID- 10673113 TI - Role of tapering in aortic wave reflection: hydraulic and mathematical model study. AB - Pressure and flow have been measured simultaneously at six locations along the aorta of an anatomically correct 1:1 scale hydraulic elastic tube model of the arterial tree. Our results suggest a discrete reflection point at the level of the renal arteries based on (i) the quarter-wavelength formula and (ii) the comparison of foot-to-foot (c(ff)) and apparent phase velocity (c(app)). However, separation of the pressure wave into an incident and reflected wave at all six locations indicates continuous reflection: a reflected wave is generated at each location as the forward wave passes by. We did a further analysis using a mathematical transmission line model with a simple tapering geometry (length 50 cm, 31 and 11 mm proximal and distal diameter, respectively) for a low (0.32 ml/mmHg), normal (1.6 ml mmHg) and high (8 ml/mmHg) value of total arterial compliance. Using the quarter-wavelength formula, a discrete reflection point is found at x = 33 cm, the level of the renal arteries, independent of the value of total compliance. However, local analysis comparing c(ff) and c(app) does not reveal a marked reflection site, and the analysis of incident and reflected waves merely suggests a continuous reflection. We therefore conclude that the measured in vivo aortic wave reflection indices are the result of at least two interacting phenomena: a continuous wave reflection due to tapering, and local reflections arising from branches at the level of the diaphragm. The continuous reflection is hidden in the input impedance pattern. Using the quarter-wavelength formula or the classical wave separation theory, it appears as a reflection coming from a single discrete site, confusingly also located at the level of the diaphragm. Therefore, the quarter-wavelength formula and the linear wave separation theory should be used with caution to identify wave reflection zones in the presence of tapering, i.e., in most mammalian arteries. PMID- 10673114 TI - Three-dimensional finite element analysis of the human temporomandibular joint disc. AB - A three-dimensional finite element model of the articular disc of the human temporomandibular joint has been developed. The geometry of the articular cartilage and articular disc surfaces in the joint was measured using a magnetic tracking device. First, polynomial functions were fitted through the coordinates of these scattered measurements. Next, the polynomial description was transformed into a triangulated description to allow application of an automatic mesher. Finally, a finite element mesh of the articular disc was created by filling the geometry with tetrahedral elements. The articulating surfaces of the mandible and skull were modeled by quadrilateral patches. The finite element mesh and the patches were combined to create a three-dimensional model in which unrestricted sliding of the disc between the articulating surfaces was allowed. Simulation of statical joint loading at the closed jaw position predicted that the stress and strain distributions were located primarily in the intermediate zone of the articular disc with the highest values in the lateral part. Furthermore, it was predicted that considerable deformations occurred for relatively small joint loads and that relatively large variations in the direction of joint loading had little influence on the distribution of the deformations. PMID- 10673115 TI - Quantifying the strain history of bone: spatial uniformity and self-similarity of low-magnitude strains. AB - We hypothesize that when a broad spectrum of bone strain is considered, strain history is similar for different bones in different species. Using a data collection protocol with a fine resolution, mid-diaphyseal strains were measured in vivo for both weightbearing and non-weightbearing bones in three species: dog, sheep, and turkey, with strain information collected continuously while the animals performed their natural daily activities. The daily strain history was quantified by both counting cyclic strain events (to quantify the distribution of strains of different magnitudes) and by estimating the average spectral characteristics of the strain (to quantify the frequency content of the strain signals). Counting of the daily (12-24 h) strain events show that large strains (> 1000 microstrain) occur relatively few times a day, while very small strains (< 10 microstrain) occur thousands of times a day. The lower magnitude strains (< approximately 200 microstrain) are found to be more uniform around the bone cross section than the higher magnitude, peak strains. Strain dynamics are found to be well described by a power-law relationship and exhibit self-similar characteristics. These data lead to the suggestion that the organization of bone tissue is driven by the continual barrage of activity spanning a wide but consistent range of frequency and amplitude, and until the mechanism of bone's mechanosensory system is fully understood, all portions of bone's strain history should be considered to possibly play a role in bone adaptation. PMID- 10673116 TI - How do changes to plate thickness, length, and face-connectivity affect femoral cancellous bone's density and surface area? An investigation using regular cellular models. AB - Models of regular cellular-solids representing femoral head 'medial group' bone were used to (1) compare thickness data for plate-like and beam-like structures at realistic surface areas and densities; (2) test the validity of a standard formula for trabecular thickness (Tb.Th); and (3) study how systematic changes in cancellous bone thicknesses, spacing, and face-connectivity affect relative density and surface area. Models of different face-connectivities, produced by plate removal from the unit cell, were fitted to bone density and surface area data. The medial group bone was anisotropic: the supero-inferior (SI) direction was the principal direction for bone plate alignment and the plane normal to this had the largest number of bone/void intersections per unit line length (P(I)). A comparison of boundary perimeter per unit area data, in planes normal to SI, with surface area data placed the medial group bone between prismatic structures in which walls are parallel to one principal direction and isotropic structures. Selective removal of plates from a closed-cell model produced a similar result. For the same relative density and surface-area, plate-like models had significantly thinner cross-sections than beam-like models. The formula for Tb.Th produced overestimates of model plate thickness by up to 20% at realistic femoral cancellous densities. Trends in data on surface area to volume ratio and density observed on sampled medial group bone could be simulated by plate thickness changes on models of intermediate face-connectivity (approximately 1.5) or by plate removal from models with relatively thick and short (low aspect-ratio) plates. The latter mechanism is unrealistic for it resulted in beam-like structures at low 'medial group' densities, an architecture unlike the predominantly plate-like bone in the sample. PMID- 10673117 TI - The effects of damage and microcracking on the impact strength of bone. AB - Microcracking has been shown to occur when bone is 'damaged' as shown by a loss of stiffness. The effect on bone's toughness of the types of damage produced at low losses of stiffness are not known. We loaded bovine bone specimens in bending and tension to stiffness losses of up to 27%, and examined the microcracking produced. The tensile specimens had diffuse arrays of microcracks of 2-20 microm in length, characteristic of tensile loading, on all surfaces. The bending specimens showed tensile microcracking on the tensile surface and characteristic long, straight, cross-hatched compression cracks on the compressive surface. Specimens were then broken in impact. Those that had been damaged in bending were divided into two groups, in one group the part of the specimen which had undergone compression damage was placed in tension, and in the other group the tensile damage was placed in tension. Tensile damage loaded in tension did not reduce the bone's energy-absorbing ability in impact until a modulus reduction of over 20%. However compression damage loaded in tension did severely reduce the bone's energy absorption capabilities (by an average of about 40%). PMID- 10673118 TI - Modeling the stability of the human glenohumeral joint during external rotation. AB - An analytical model of the human glenohumeral joint was developed to predict glenohumeral kinematics and investigate how the glenohumeral capsule and articular contact between the humeral head and the glenoid stabilize the joint. This was performed during a simulation of an apprehension clinical exam or the cocked phase of throwing, when the humerus is susceptible to anterior instability or dislocation. Contact between the joint surfaces was modeled using a deformable articular contact method and the capsule was modeled as five elements with the ability to wrap around the surface of the humeral head. Experimental measurements (Novotny et al., Journal of Shoulder and Elbow surgery, 1998, 7, 629-639) provided geometric data from four in vitro specimens and kinematic results to validate model predictions. Material properties were taken from the literature. An equilibrium approach was used with the forces and moments produced by the ligaments and surface contact balanced against those applied externally to the humerus during external rotation of the abducted and extended humerus. The six equilibrium equations were solved for the position and orientation of the humerus. The center of the humeral head translated posteriorly and superiorly with external rotation. Model predictions for translational and rotational ranges of motion were not significantly different from experimental findings; however, at individual moment increments, the model underestimated the external rotation and overestimated the superior-inferior position of the humerus relative to the glenoid. The anterior band of the inferior glenohumeral ligament increased in tension with external rotation, while the axillary pouch and posterior band decreased in tension. Contact area, stress and force increased with external rotation and the contact area moved posteriorly and inferiorly around the rim of the glenoid. The model results provide information on how the relationship between the ligament element tensions and contact forces may act to avoid glenohumeral instability. PMID- 10673119 TI - Study on blood constitutive parameters in different blood constitutive equations. AB - A method is proposed to study the steady blood constitutive equation (BCE). With the method several BCEs are tested and Quemada, Bi-exponent and K-L equations are found to be well in agreement with the hemorheological characteristics of human and canine blood and these equations may be well used in hemorheology and hemodynamics. In addition, the potential clinical applications of some blood constitutive parameters (BCPs) are discussed. It is found that the hematocrit can significantly affect the BCPs: K(A) = alpha2/alpha1, Q(A) = eta0/eta(infinity), R(A) = etaA/eta(e) - 1, which means relative viscosities and they may be related to RBC aggregative level. Moreover, we also show that the parameters are related to the RBC aggregative strength and level. PMID- 10673120 TI - A telemetry-based device to determine the force-displacement behaviour of materials in high impact loading situations. AB - Meaningful testing of stab resistant body armour requires the use of realistic body tissue simulants. A device for the determination of the force-displacement behaviour of materials in high impact loading situations has been developed for the testing of such simulants. Force measurement is achieved with the use of electrical foil strain gauges applied to a cylindrical load cell. A piezo resistive accelerometer (+/- 500 g) is used to calculate the displacement of the device through double integration of its signal, with the impact velocity used as a boundary condition. The signals from the strain gauge circuit and the accelerometer are sampled at 2500 Hz. The data are transmitted to a receiver via telemetry using a 418 MHz FM transmitter and from the receiver to a laptop PC via the serial port. Calibration of the device is described and sample results showing forces up to 2500 N and displacements up to 0.04 m are presented. PMID- 10673121 TI - Constitutive modelling of abdominal organs. AB - Abdominal organs are very susceptible to trauma. In order to protect them properly against car crash and other impact consequences, we need to be able to simulate the abdominal organ deformation. Such simulation should account for proper stress-strain relation as well as stress dependence on strain rate. As the step in this direction, this paper presents three-dimensional, non-linear, viscoelastic constitutive models for liver and kidney tissue. The models have been constructed basing on in vivo experiments conducted in Highway Safety Research Institute and the Medical Centre of The University of Michigan (Melvin et al., 1973). The proposed models are valid for compressive nominal strains up to 35% and fast (impact) strain rates between 0.2 and 22.5 s(-1). Similar models can find applications in computer and robot assisted surgery, e.g. the realistic simulation of surgical procedures (including virtual reality) and non-rigid registration. PMID- 10673122 TI - In vivo measurement-based estimations of the moment arm in the human tibialis anterior muscle-tendon unit. AB - The aim of this study was to estimate the moment arm of human tibialis anterior (TA) muscle-tendon unit at rest and during isometric dorsiflexion maximum voluntary contraction (MVC) from in vivo sagittal-plane magnetic resonance (MR) and ultrasound scans. Two methods were employed, both of them based on the assumption that the ankle joint complex and TA muscle-tendon unit operate in the sagittal plane. Using method A, moment arms were obtained from MR scans of the foot by measuring the perpendicular distance between a moving centre of rotation in the talo-crural joint and the TA tendon action line. Using method B, moment arms were calculated from the ratio of TA tendon displacement, which was estimated from a planimetric muscle model using pennation angles and muscle thickness measured by ultrasonography, to the tibial rotation around the talus, which was measured from the foot MR scans. Using either of the two methods at rest, the estimated TA moment arm decreased from approximately 4.5 to approximately 2.9 cm in the transition from dorsiflexion to plantarflexion. Using method A, moment arms during MVC were larger by 0.9-1.5 cm (33-44%, P < 0.01) than the respective resting estimations. In contrast, no difference (P > 0.05) was found between the resting and MVC moment arm estimations of method B. Limitations in the oversimplified musculoskeletal model used raise questions for the validity of both method estimations. PMID- 10673123 TI - Characterization of load-length-velocity relationships of nine different skeletal muscles. AB - A three-dimensional characterization of muscle load, length and velocity was obtained from nine muscles in the cat's hind limb through contractions where the muscles shortened against inertial-gravitational loads. A model based on the load length characteristic and second-order dynamics describes shortening velocity related to load and length under these conditions. We conclude that this model describes well contraction velocity as function of length and load under inertial gravitational load conditions, with correlation coefficients higher than 0.9 in most of the tested muscles. PMID- 10673124 TI - Thresholds for step initiation induced by support-surface translation: a dynamic center-of-mass model provides much better prediction than a static model. AB - The need to initiate a step in order to recover balance could, in theory, be predicted by a static model based solely on displacement of the center of mass (COM) with respect to the base of support (BOS), or by a dynamic model based on the interaction between COM displacement and velocity. The purpose of this study was to determine whether the dynamic model provides better prediction than the static model regarding the need to step in response to moving-platform perturbation. The COM phase plane trajectories were determined for 10 healthy young adults for trials where the supporting platform was translated at three different acceleration levels in anterior and posterior directions. These trajectories were compared with the thresholds for step initiation predicted by the static and dynamic COM models. A single-link-plus-foot biomechanical model was employed to mathematically simulate termination of the COM movement, without stepping, using the measured platform acceleration as the input. An optimization routine was used to determine the stability boundaries in COM state space so as to establish the dynamic thresholds where a compensatory step must be initiated in order to recover balance. In the static model, the threshold for step initiation was reached if the COM was displaced beyond the BOS limits. The dynamic model showed substantially better accuracy than the static model in predicting the need to step in order to recover balance: 71% of all stepping responses predicted correctly by the dynamic model versus only 11% by the static model. These results support the proposition that the central nervous system must react to and control dynamic effects, i.e. COM velocity, as well as COM displacement in order to maintain stability with respect to the existing BOS without stepping. PMID- 10673125 TI - Comments on 'A multi-phase optimal control technique for the simulation of a human vertical jump'. PMID- 10673126 TI - The effectiveness and safety of brimonidine as mono-, combination, or replacement therapy for patients with primary open-angle glaucoma or ocular hypertension: a post hoc analysis of an open-label community trial. Glaucoma Trial Study Group. AB - The purpose of this study was to determine the effectiveness and safety of brimonidine 0.2% (Alphagan, Allergan Inc., Irvine, CA) as mono-, combination, or replacement therapy for reducing intraocular pressure (IOP) in patients with primary open-angle glaucoma or ocular hypertension. The study method was an open label, comparative clinical evaluation involving 2335 patients. During the 2 month trial, data were collected at baseline (visit 1), month 1 (visit 2), and month 2 (visit 3). Various parameters were evaluated, including glaucoma medications (visit 1), IOP (visit 1-visit 3), and adverse events. A subset cohort of 1254 patients was selected that met specific study criteria. Data from these 1254 patients were used to evaluate adverse events and the change in IOP from visit 1 to visit 3. Patient data were grouped according to specific drug regimen, and drug regimens were categorized into supergroups of mono-, combination, and replacement therapy. The results of the study revealed that the overall mean change in IOP for 1) monotherapy (n = 240) was -5.07 mm Hg (-20.2%), 2) combination therapy (n = 554) was -4.01 mm Hg (-16.9%), 3) replacement therapy (n = 460) was -2.33 mm Hg (-9.8%), and 4) overall (n = 1254) was -3.59 mm Hg ( 14.9%) (p < 0.001 for all changes). Overall, 6.0% of the subjects reported adverse events, with no hypersensitivity or unexpected systemic or ocular adverse events. Eighty-five percent (85%) of clinicians rated brimonidine as "good" to "excellent". In conclusion, brimonidine is safe and effectively lowers IOP when used as mono-, combination, or replacement therapy as observed in a large community population. PMID- 10673127 TI - Aqueous humor dynamics in monkeys with laser-induced glaucoma. AB - This study determines the effects of laser-induced glaucoma on aqueous humor dynamics of 18 cynomolgus monkeys. Baseline measurements of 12 monkeys included intraocular pressure (IOP) by pneumatonometry, aqueous flow by fluorophotometry and outflow facility by tonography. Beginning 4 to 14 days later, the trabecular meshwork of one eye was treated repeatedly with laser photocoagulation until elevated IOP was induced. Thirty-six to 75 days after the last laser treatment, all measurements were repeated. Between 1.7 and 11.4 years after laser treatment, the same 12 monkeys plus 6 additional monkeys underwent IOP and aqueous flow measurements. In addition, outflow facility was determined with fluorophotometry, and uveoscleral outflow was both calculated (n=18) and measured with an intracameral tracer (n=7). In glaucoma eyes compared to control eyes (n=12), IOP was increased (p<0.04) by at least 8 mmHg at Time 1 (1 to 3 months) or Time 2 (3 to 4 years) after laser treatment; aqueous flow was reduced (p=0.0007) by 46% at Time 1 but returned to baseline levels at Time 2; tonographic outflow facility was reduced (p=0.0008) by 71% at Time 1. In lasered eyes compared to control eyes, fluorophotometric outflow facility was reduced (p=0.0008; n=18) by 63%, and uveoscleral outflow was increased (p<0.05), whether calculated or measured with tracers at least 1 year after laser treatment. The increased IOP in monkeys with laser-induced glaucoma was caused by a sustained reduction in outflow facility. The uveoscleral outflow increase was not enough to prevent the rise in IOP. PMID- 10673128 TI - Topical rivastigmine, a selective acetylcholinesterase inhibitor, lowers intraocular pressure in rabbits. AB - Non-selective acetylcholinesterase (AChE) inhibitors are known hypotensive agents. The purpose of the present investigation was carried out to ascertain whether rivastigmine, a selective carbamate-type inhibitor of AChE, which inhibits selectively an isoform of this enzyme found almost exclusively in the central nervous system, is able to depress the intraocular pressure (IOP) in normotensive rabbits. IOP was monitored with a TonoPen XL in conscious adult rabbits before and hourly up to 8 hr after administration of the drug. Baseline measurements without treatment and after one single topical application of rivastigmine [1% (n=8); 2% (n=4); and 5% (n=6)] to the right eye and of the vehicle alone to the left one were performed. Rivastigmine reduced the IOP of treated eyes significantly (p<0.05) in a dose-independent manner. Maximal effects of 23.2% (5% rivastigmine), 19.6% (2% rivastigmine) and 15.2% (1% rivastigmine) were achieved 1, 3 and 5 hr after application of the drug. A non-significant reduction of IOP in the contralateral eye was also observed. Rabbits evidenced no signs of discomfort after administration of rivastigmine. No conjunctival discharge or other signs of drug related local toxicity were found. Rivastigmine, a selective antagonist of AChE, lowers IOP significantly and may thus be of potential use in glaucoma therapy. PMID- 10673129 TI - The application of water drinking test on the evaluation of trabeculectomy patency. AB - The water drinking test (WDT) was once frequently used as a diagnostic tool for glaucoma, but not so often nowadays. In this study, we investigated the potential use of the WDT on the evaluation of trabeculectomy patency. Twenty age-matched volunteers and thirty-six glaucoma patients who were to receive trabeculectomy procedure were enrolled in this study. The WDT was given to the volunteers once and to all glaucoma patients before undergoing trabeculectomy and at certain intervals after the procedure. The WDT was performed in a standard manner. We defined four parameters after performing the WDT: Initial Pressure [IP, intraocular pressure (IOP) level before the WDT]; Slope of Ascending Trend (SOAT, the slope between baseline IOP and the highest IOP level); Peak Pressure (PP, the highest IOP level during the WDT); and End Pressure (EP, the IOP level after the WDT). It was found that the results of the WDT and trabeculectomy patency were strongly correlated. The four parameters in success and failure cases were significantly different at the last follow up: IP: (15.2 +/- 3.6 vs. 25.3 +/- 6.4, p<0.01); SOAT: (0.9 +/- 0.3 vs. 1.8 +/- 1.2, p<0.01); PP: (19.2 +/- 6.4 vs. 39.5 +/- 12.2, p<0.01); EP: (15.5 +/- 4.8 vs. 29.4 +/- 8.2, p<0.01). Thus, it was observed in this study that WDT was not only easy and safe to perform, but also valuable in evaluating the patency of trabeculectomy. PMID- 10673130 TI - Efficacy of latanoprost as an adjunct to medical therapy for residual angle closure glaucoma after iridectomy. AB - Residual primary angle-closure glaucoma (PACG) after iridectomy is an important issue among Asians, especially Chinese. In this study, we tested the effectiveness of latanoprost as an intraocular pressure (IOP) lowering agent in cases of residual PACG. Twenty-six eyes of 26 PACG patients with persistently elevated IOP after iridectomy, despite treatment with conventional IOP lowering drugs (beta blockers and pilocarpine) were included. Latanoprost 0.005%, one drop daily, was added adjunctively to all eyes. Measurement of IOP at baseline and after the start of treatment with latanoprost indicated a significant IOP reduction. The IOP decreased by about 21% (p < 0.005) during the first 3 months, and showed a reduction of about 36% at the end of 1 year. At the 1-year follow up, the IOP was well controlled (below 20 mmHg) in all eyes. These findings show that, in combination with beta blockade and pilocarpine, latanoprost can ameliorate residual PACG after iridectomy and could potentially forestall the need for further therapeutic intervention. PMID- 10673131 TI - Circadian variations of prostaglandins in the rabbit aqueous humor. AB - Forty-eight young adult New Zealand albino rabbits were entrained to a daily 12 hr light and 12-hr dark cycle. Under a constant dark environment, rabbits were sacrificed at 4-hr intervals, beginning at 2 hr before the accustomed lights-on period. Eight rabbits were used for each of the 6 time points within 24 hr. Aqueous humor and vitreous humor samples were collected. Samples from one eye of each rabbit were assayed for prostaglandins D2, E2, and F2alpha by enzyme immunoassay. Total protein concentrations in the samples were also determined. Concentrations of prostaglandins D2 and F2alpha in the aqueous humor were found to vary in a consistent pattern. The prostaglandin F2alpha concentration increased in the early subjective light period, remained high in this period, and decreased in the subjective dark period. The prostaglandin D2 concentration was elevated only in the early subjective light period. No significant change of aqueous humor prostaglandin E2 level was found. Total protein concentration in the aqueous humor varied in a similar pattern as the aqueous humor prostaglandin F2alpha, indicating a close association between the permeability of protein across the blood-aqueous barrier and the endogenous prostaglandin F2alpha level. In the vitreous humor, concentrations of all three prostaglandin subtypes remained unchanged, as did the total protein concentration. However, vitreous humor concentrations of prostaglandin D2 and F2alpha were significantly higher than their concentrations in the aqueous humor. PMID- 10673132 TI - Effect of nitric oxide synthesis inhibition on post-occlusive choroidal blood flow in rats. AB - Experiments were designed to study involvement of nitric oxide on vascular responses to ocular ischemia in the anesthetized rat. Anterior choroidal blood flow was measured using laser-Doppler flowmetry. In some experiments, cerebral cortical blood flow also was measured. Ischemia was produced by either occlusion of the cephalic blood supply or more locally via a ligature tightened around the eye stalk. Arterial blood pressure and choroidal blood flow was continuously measured before, during and after a 20 min ischemic challenge. Both methods of ischemia reduced choroidal blood flow (>90%) with no consistent ocular hyperemia seen upon reperfusion. No significant differences in response pattern between the two ischemia techniques were apparent. Treatment with the non-selective inhibitor of nitric oxide (L-NAME 2 mg/kg, i.v.) did not alter either basal choroidal blood flow or the pattern of reperfusion. A larger dose of L-NAME (50 mg/kg, i.v.) reduced both basal choroidal blood flow and the final reperfusion level (most likely due to continued depression of the basal ocular choroidal blood flow). Neither D-NAME nor the neuronal nitric oxide synthase inhibitor, 7-nitroindazole, altered basal anterior choroidal blood flow or the reperfusion pattern seen after reperfusion. The results confirm our previous observations that inhibition of endothelial nitric oxide lowers. basal choroidal blood flow in the rat eye. However, in contrast to the cerebral circulation where L-NAME greatly attenuates initial reperfusion to the cerebral cortex, inhibition of nitric oxide synthase does not appear to notably further influence anterior choroidal reperfusion. PMID- 10673133 TI - Cell signaling in bovine ciliary epithelial organ culture. AB - The ciliary epithelium secretes aqueous humor, an intraocular fluid whose production is regulated in part by transmembrane signaling pathways including those mediated by G protein-coupled receptors. Many drugs, such as beta adrenergic receptor (AR) antagonists and alpha2-AR agonists, are used to lower intraocular pressure by presumably decreasing fluid transport across this epithelium. Hence, our purpose was to establish a ciliary epithelial organ culture system suitable for the study of cell signaling pathways. A trypsin mediated dissection method was established to isolate bovine ciliary epithelial sheets. These sheets were cultured in a 5% CO2 incubator. The quality was assessed by light microscopy, by protein analysis, and by the evaluation of epinephrine-mediated phosphoinositide turnover. The cultured epithelial explants were viable as evidenced by minimal trypan blue staining. The explants were composed primarily of nonpigmented cells and some pigmented cells, but no other ciliary body tissues were present on histology. Membrane preparations showed proteins with a distribution from 31 to 116 kDa. Epinephrine caused a dose dependent increase in [3H]inositol phosphates (InsPs) accumulation with a maximal increase of two- to three-fold over basal levels. This epinephrine-mediated increase was inhibited by prazosin. We established an organ culture system of isolated bovine ciliary epithelium suitable for the study of transmembrane signaling pathways. PMID- 10673134 TI - Antibiotic treatment of orbital cellulitis: an analysis of pathogenic bacteria and bacterial susceptibility. AB - The proper choice of effective antibiotics is a mainstay for the treatment of orbital cellulitis. The lack of native data regarding the microorganism causing the infection and its antibiotic sensitivity prompted us to conduct this study. We retrospectively collected 29 cases of orbital cellulitis admitted to Chung-Ho Memorial Hospital of Kaohsiung Medical College from January 1994 to September 1998. The effectiveness of antibiotics with bacterial susceptibility was analyzed. Of the 29 cases, fifteen were male and fourteen female. The patients ranged in age from 7 months to 79 years (mean, 37.6 years). Sinusitis (9 cases, 31.0%) is the most common etiology. Fourteen cases received both medical and surgical treatments. Eighteen cases had purulent discharge from the infection areas sent for culture isolation of the microorganism. The culture positive rate was 50% (9 in 18 cases). The Staphylococcus aureus (5 cases) was the most common pathogen. The bacterial susceptibility test showed drug resistance of 100% for penicillin G (seven out of seven cases; 7/7), 100% for ampicillin (10/10), and 0% for amikacin (0/3) and vancomycin (0/7). Penicillin and ampicillin are not effective for those isolated bacteria. Oxacillin and gentamicin, frequently used in first line treatment, might encounter drug resistance in some cases. Amikacin and vancomycin, without any resistance in bacterial susceptibility tests, could be used in vision-threatening, critical, and intractable cases. PMID- 10673135 TI - Effects of interleukin-1 blockers on corneal fibroblast proliferation in vitro and ocular inflammation in vivo. AB - The success of keratorefractive surgical procedures is limited by the wound healing process in the corneal stroma. The proliferation and matrix synthesis of corneal stromal fibroblasts is the central element of the wound healing process that is triggered by an initial inflammation. In order to develop new therapeutic strategies to reduce wound healing intensity, we investigated the effect of newly synthesized interleukin-1 (IL-1) blockers on the proliferation of cultured rabbit corneal fibroblasts and the ocular inflammation induced by IL-1. It was found that the addition of IL-1 blockers, such as CK-135 to CK-145, led to a dose dependent inhibition of cell proliferation after 24, 48 and 72 hr of incubation. The isotope incorporation study showed that the syntheses ofDNA and mRNA were suppressed whereas that of protein was enhanced or unaffected. These compounds also demonstrated a potent anti-inflammation action in the rat uveitis model. Our results indicate that CK (Chiou-Kumamoto) compounds may be valuable therapeutic agents for the prevention of postoperative complications after corneal keratorefractive surgical procedures. PMID- 10673136 TI - Kinetic models for normal and impaired growth. AB - Although microkinetic models are expected to play in the future the same role that macrokinetic models have played in the past, classic mechanistic growth functions are still worthy of study and may still provide insight into auxological problems. The rather rigid shape of macrokinetic models may ignore many interesting fluctuations of growth velocity, but a strong structure allows a robust estimate of growth kinetics even in the case of growth profiles which are largely incomplete, as those derived from current clinical records. In any case, the too simplified shape of these models may be adjusted, to some extent, by adding some unstructured smooth function of residuals which takes into account minor aspects of growth (such as slight spurts during childhood), which cannot be detected in an individual profile because of random errors and inadequate number of observations. This paper recalls the reasons why growth models are useful, analyses briefly the structure and the characteristics of the two fundamental human growth functions, i.e. triple-logistic and PB1, and shows how the use of PB1 model may be extended also to impaired growth, e.g. in girls with Turner syndrome. In this regard, the use of the same model for normal and pathological growth offers the important advantage that differences between growth patterns are not confounded with differences between models. PMID- 10673137 TI - Relationships between blood pressure, anthropometric characteristics and blood lipids in high- and low-altitude populations from Central Asia. AB - We studied the relationships between blood pressure, anthropometric characteristics and blood lipids in 72 low altitude (LA) Uighurs (600 m), 91 LA Kirghizs (900 m), 117 medium altitude (MA) Kazakhs (2100 m) and 94 high altitude (HA) Kirghizs (3200 m). All subjects were male and had a similar age (p = ns, ANOVA; range for all 374 subjects: 18-66 yr). Body weight (Wt), body mass index (BM1) and the sum of four skinfolds (4SF) were significantly lower in HA-Kirghizs than the remaining groups (p < 0.0005, p < 0.0005 and p < 0.05 respectively, ANOVA). However, no difference was found in body fat distribution as detected by waist:hip circumference (WHR) and triceps:subscapular skinfold ratios (TSR; p = ns, ANOVA). Stage 1 hypertension was detected in 18% of LA-Uighurs, 2% of LA Kirghizs, 4% of MA-Kazakhs and 1% of HA-Kirghizs; stage 2 hypertension was detected in 2% of LA-Uighurs and none of the remaining groups; no subject had stage 3 hypertension (The Joint National Committee on Prevention. Detection, Evaluation and Treatment of High Blood Pressure 1997). Blood cholesterol (CH) and triglycerides (TG) did not differ between groups (p = ns, ANOVA). The relationships between systolic (SBP) or diastolic (DBP) blood pressure and age, Wt, BMI, 4SF, WHR, TSR, CH and TG were independent from altitude (p = ns, ANCOVA). In the pooled sample (n = 374), age explained 1 and 3% of SBP (p < 0.05) and DBP (p < 0.005) variance respectively, Wt was the best predictor of SBP and DBP explaining 11 and 10% of their variance respectively (p < 0.0001) and CH explained 5% of DBP variance (p < 0.0001). In conclusion, hypertension is more frequent in LA- than MA- and HA-subjects from Central Asia. However, anthropometric characteristics and blood lipids do similarly contribute to explain blood pressure in these subjects. PMID- 10673138 TI - High heterogeneity of apolipoprotein E gene frequencies in South American Indians. AB - The apolipoprotein E (APOE) polymorphism was investigated in 186 individuals from six South American Indian tribes, and the results integrated with those previously presented for this ethnic group. The three APOE alleles commonly reported in other populations were also observed in South Amerindians with a highly heterogeneous distribution. As in other populations, APOE*3 was the most common allele (51-98%) followed by APOE*4 (2-47%). These two isoforms were identified in all tribes, but APOE*2 was observed among the Wai Wai (2%) and Mataco (4%) only. No previous indications of inter-ethnic admixture were observed among the Wai Wai, but the introduction of this allele among the Mataco through non-Indian sources cannot be excluded. PMID- 10673139 TI - Comparison of two methods for transforming height and weight to normality. AB - I assessed the success of the Box-Cox power transformation and the shifted log transformation for transforming length/height and weight to Normality. Retrospectively collected data for full-term, healthy 1143 boys and 1162 girls amounted to over 45000 recordings. Of the children, 48%, born during 1959-1961, were followed from birth to age 19.0, and the rest, born during 1969-1971, until age 12.5. The individual growth curves were smoothed in order to obtain reliable interpolated estimates of measurements at fixed ages and to avoid age grouping. Height is fairly Normal, even during puberty, and weight can be transformed close to Normality, the Box-Cox transformation being preferable to the shifted log transformation. Occasionally remaining leptokurtosis in the transformed weight can apparently be disregarded when focusing on the extreme (1st, 2.5th, 97.5th and 99th) centiles. The 95% confidence intervals for the two transformation parameters are wide, and may even explode for the shift parameter. The shapes, although not the levels, of the curves joining the estimates are similar for boys and girls, showing a 2-year difference in maturation. Improved screening for abnormal growth, based on the assumption of Normality, after a transformation, seems possible. PMID- 10673140 TI - Quantifying the 'appleness' or 'pearness' of the human body by subcutaneous adipose tissue distribution. AB - To quantify subcutaneous adipose tissue topography (SAT-Top) describing individual SAT distribution for a subject or even a group we measured subcutaneous adipose tissue thickness at 15 specified body sites of 303 healthy women aged 20-69 yrs and 20 women with proven non-insulin-dependent diabetes mellitus (NIDDM) by the optical device 'LIPOMETER'. The type of upper-body- and lower-body-fat pattern, (apples or pears), was determined by factor analysis of the data. Upper body sites were highly loaded in factor 1, whereas factor 2 included highly loaded body sites from the lower extremities. For an individual, factor 1 scores > factor 2 scores, was described as an 'apple'-type, while factor 2 > factor 1 was described as a 'pear'-type. We found about 80% 'pears' and 20% 'apples' in 20-29 year olds and 20% 'pears' and 80% 'apples' in 60-69 year old women. Women with NIDDM tended to be 'super-apples'. SAT-Top provides a useful differentiation between apples and pears and we recommend this approach as a screening method. PMID- 10673141 TI - The importance of considering biological maturity when assessing physical fitness measures in girls and boys aged 10 to 16 years. AB - It is widely considered that biological maturity influences physical fitness test performance, children can be advantaged/disadvantaged in physical fitness tests by being more or less mature than counterparts of the same chronological age. The current study sought to investigate the effect sexual maturity had upon performance in physical fitness tests. A cross-sectional study involving 161 girls and 152 boys was carried out. Each subject was assessed for stature, mass, self-assessment of sexual maturity, vertical jump, hand grip strength and the 20 m shuttle run test, all procedures were standardized. Spearman's rank correlation coefficients were developed to assess the relationship between maturity and physical fitness measures. ANCOVA inferential statistics were performed to investigate if performance in physical fitness tests differed between children of different sexual maturity stages irrespective of mass and stature. Significance was set at p < 0.05. Stage of sexual maturity was significantly correlated with all physical fitness measures (boys: r=0.56 to 0.73; girls: r=0.24 to 0.46). ANCOVA revealed that when stature and mass were taken into account significant differences were evident between sexual maturity stages in boys but not girls. This suggests that increases in mass and stature are primarily responsible for variation in girls' physical performance throughout maturation, whereas in boys there are some qualitative differences in performance due to other factors. It was concluded that sexual maturity has a large influence on physical fitness measures in boys but less effect in girls. Rating of physical fitness, particularly for boys should take into account biological maturity. PMID- 10673142 TI - Growth pattern of the Sugalis--a tribal population of Andhra Pradesh, India. AB - With a view to assess physical growth, a cross-sectional study was made on 1565 Sugali children (854 boys and 711 girls), aged 1 to 20 years. Anthropometric measurements included height, weight, upper arm circumference, biacromial diameter, biiliocristal diameter, chest circumference, head circumference and skinfold measurements at triceps, subscapular, suprailiac and medial calf. All anthropometric measurements except skinfold measurements exhibit uniform increase with age in both the sexes. A gradual increase in four skinfold measurements is observed with age in the case of girls, whereas slight decrease is observed in the case of the boys. The Sugali boys and girls are shorter and lighter than well to-do Indian standards. The median heights and weights of Sugali boys and girls fall below the 5th percentile of NCHS standards. Finally, the results were discussed with a comparative view point. PMID- 10673143 TI - Phylogenetic relationships of a class of hominoid-specific retro-elements (SINE R) on human chromosomes 7 and 17. AB - SINE-R elements are a class of retroposon derived from the human endogenous retrovirus HERV-K that has been active in hominoid evolution and may include some members that are Homo sapiens specific. Both SINE-R elements and the HERV-K class of element have potential relevance to recent genome change. Here we report on sequences in the SINE-R class that can be detected on human chromosomes 7 and 17 and compare them with sequences that we have previously reported on the X chromosome and in hominoid primates. The retroposons on chromosomes 7 and 17 showed a high degree of sequence homology (88-96%) with other human retroposons (SINE-R.C2, 11, 14, 19, and HS307/HS408). Phylogenetic analysis using the neighbour-joining method revealed that SINE-R-type retroposons on chromosomes 7 and 17 were inter-related with those of hominoid primates, suggesting that various sub-classes of these retroposons have been evolving independently during hominoid evolution. One element (17-11) on chromosome 17 shares one hundred per cent identity with a 7-11 element on chromosome 7 which suggests either recent transposition or a chromosomal translocation. Thus further investigation of the chromosomal locations of SINE-R elements that together with the HERV-K LTR sequence from which they are derived have the capacity to influence the function of neighbouring cellular genes may be expected to clarify the potential role of these elements in recent hominoid evolution. PMID- 10673144 TI - Mitochondrial-nuclear interactions and lifespan control in fungi. AB - In fungi, mitochondrial-nuclear interactions are part of a complex molecular network involved in the control of aging processes. The generation of reactive oxygen species at the mitochondrial respiratory chain plays a major role in this network. Mitochondrial DNA instabilities, which are under the control of nuclear genes, affect the generation of reactive oxygen species and modulate the rate of aging. As mitochondria become dysfunctional, they transduce signals to the nucleus and induce the expression of a set of nuclear genes, a process termed retrograde regulation. Molecular data are emerging which suggest that retrograde regulation is involved in lifespan control. PMID- 10673145 TI - Biomarkers of immunosenescence within an evolutionary perspective: the challenge of heterogeneity and the role of antigenic load. AB - Under an evolutionary perspective, antigens can be considered nothing else than chronic stressors that constituted the major selective pressure for immune system emergence and evolution. In this review, recent data are discussed under the hypothesis that human immunosenescence is the consequence of the continuous attrition caused by chronic antigenic overload/stress. The advantage of this theoretical approach is that a unifying hypothesis is proposed, which tries to fill in the current gap between the conceptualizations concerning the mechanisms which counteract aging and favor longevity in invertebrates and vertebrates. The hypothesis is that the immune system is, at a higher level of biological organization and complexity, the counterpart of the anti-stress response network identified in invertebrates as the major determinant of survival. We argue that some of the most important characteristics of immunosenescence, i.e. the accumulation and the clonal expansion of memory and effector T cells, the reduction/exhaustion of naive T cells, and the shrinkage of T cell repertoire, are compatible with this assumption. Thus, immunosenescence can be envisaged as a global reduction of the "immunological space." Concomitantly, immunosenescence results in the progressive generation of cellular mosaicism which is the consequence of the heterogeneous replicative histories and telomere shortening of T and B cell subsets, as well as hemopoietic stem cells. Most of the parameters affected by immunosenescence appear to be under genetic control, and future research on biomarkers should address this point. On the whole, immunosenescence can be taken as a proof that the beneficial effects of the immune system, devoted to the neutralization of dangerous/harmful agents early in life and in adulthood, turn to be detrimental late in life, in a period largely not foreseen by evolution. This perspective fits with basic assumptions of evolutionary theories of aging, such as antagonistic pleiotropy. PMID- 10673146 TI - Hormone replacement in the aging male?. AB - The recent increase in the elderly population, current health trends, and awareness of age-related changes in the male endocrine system have led to discussions about the endocrine system being a "pacemaker" of male aging. Better prevention and treatment of "non-beneficial states of health" in aging men, such as generally decreased well-being and virility, increased visceral fat, osteopenia, atherosclerosis, and impaired cognitive function, is based on improved understanding of aging, particularly the significance of age-associated hormonal changes. Although not as dramatic as in women, men also show declining hormone serum concentrations with aging (Vermeulen and Kaufman, 1995). Because aging is associated with deleterious effects resembling the clinical signs of hypogonadism or states of subnormal hormone concentrations, the potential role of hormone supplementation in aging men paralleling hormone replacement therapy in postmenopausal women is the topic of discussion. The goal of hormone replacement would be to improve body composition and increase muscle strength and quality of life in men, thereby reducing mortality and morbidity. The findings so far support the need for long-term studies of hormonal supplementation in older males showing decreased hormone serum levels. Nevertheless, to date, such a use outside the context of a clinical trial is not justified. PMID- 10673147 TI - Werner helicase expression in human fetal and adult aortas. AB - Werner syndrome is a human progeroid syndrome caused by mutations at the Werner helicase locus (WRN). Progeroid features and diseases associated with aging (including arteriosclerosis) do not become apparent until after puberty. We entertained two alternative hypotheses to explain the post-pubertal onset: 1) WRN expression is induced at the time of puberty, its earlier functions being satisfied by another member of that family of helicases; and 2) it is expressed at all ages, but the phenotype of deficiency becomes apparent only after puberty. We report initial experiments consistent with the second hypothesis. Steady-state levels of WRN mRNA in aortic tissues were determined by semiquantitative reverse transcription-polymerase chain reaction. WRN mRNA was detectable as early as 49 days of gestation (the earliest available material). There was no statistically significant change in these levels between fetal and adult tissues. The presence of the WRN protein in fetal aorta was confirmed by Western analysis. This rules out the possibility that Werner syndrome phenotypes manifest after the puberty because of peripubertal induction of WRN expression. PMID- 10673148 TI - Ceroid/lipofuscin-loaded human fibroblasts show decreased survival time and diminished autophagocytosis during amino acid starvation. AB - To test whether heavy accumulation of ceroid/lipofuscin can disturb important functions of the lysosomal system, AG-1518 human fibroblasts, ceroid/lipofuscin loaded (following prolonged culture at normobaric hyperoxia) or not, were exposed to amino acid starvation. Ceroid/lipofuscin-loading resulted in decreased cellular survival. Also, there was an inverse relationship between amounts of ceroid/lipofuscin and the survival time of individual cells within the same cultures. Ceroid/lipofuscin-loaded fibroblasts displayed diminished autophagocytotic capacity, as demonstrated by electron microscopy and by treatment of cell cultures with NH4Cl (which inhibits autophagocytotic degradation by increasing intralysosomal pH) for 1 week before ensuing starvation. The latter treatment increased survival of control cells (due to deposition of nondegraded autophagocytosed material before start of starvation), but not that of ceroid/lipofuscin-loaded cells. Moreover, when NH4Cl treatment was combined with starvation, both groups of cells showed approximately the same shortened survival times, testifying to the causal relationship between diminished autophagocytosis and decreased survival of starving ceroid/lipofuscin loaded cells. We hypothesize that large amounts of undegradable ceroid/lipofuscin within the acidic vacuolar compartment may interfere with lysosomal function, resulting in poor renewal of long-lived proteins and worn-out/damaged organelles, decreased adaptability, and cell death. PMID- 10673149 TI - Phenotypic and functional characteristics of circulating monocytes of elderly persons. AB - Aging is associated with impairment of immune functions. Age-dependent alterations in T-cells are well known. Although the pivotal role of monocytes in immune regulation by their production of proinflammatory and inhibitory cytokines is acknowledged, limited information is available on monocyte changes in aging. The present study focused on phenotypic changes in circulating monocytes in elderly subjects and in the level of cytokines they produce. The results demonstrated a significant expansion of CD14dim/CD16bright circulating monocytes in elderly. In contrast, the majority of circulating monocytes of healthy young individuals were CD14bright/CD16dim. The CD14dim/CD16bright monocytes are considered to have phenotypic evidence for activation. Furthermore, significant increases of constitutive production of monocytic cytokines including interleukin (IL)-1beta. IL-1 receptor antagonist, and IL-6 by nonstimulated monocytes from elderly was also indicative of activation. This was also observed when monocytes from elderly were cultured with autologous lymphocytes. However, after stimulation, significantly lowered IL-1beta production was observed and IL-6 and IL-10 tended to be higher in the elderly. Collectively, these results indicate that monocytes of aged individuals, in contrast to a younger population exhibit in vivo activation as well as imbalanced production of cytokines. Such age related alterations in monocytes may contribute to impaired immune competence of aging. PMID- 10673150 TI - Glucocorticoid receptors in ageing rats. AB - The role of the glucocorticoid receptor (GR) in senescence was studied in rats of increasing age. Statistically significant changes in the number of GRs from rat liver were detected, whereas the affinity for the ligand triamcinolone acetonide (TA) did not change with increasing age, and was in the range of 1-2 nM. In all cases the number of receptors was lower in rats treated with hormone in vivo relative to untreated animals. In addition, we have found changes in GR activation, as measured by the binding to DNA cellulose in the mentioned age groups. Furthermore, expression of the glucocorticoid hormone (GH)-inducible gene, tyrosine amino transferase (TAT) also showed age-related alterations. We conclude that receptor function shows oscillatory changes during ageing. In addition, response to GH generally declines towards the older age. This specific periodicity in functional characteristics of the GR may reconcile conflicting results about the receptor number and properties during the ageing process, and marks particular age at which individual organism shows the highest or the lowest sensitivity to the actions of GH. PMID- 10673151 TI - 1,25-Dihydroxy-vitamin D3 (calcitriol)-dependent protein phosphorylation in rat duodenum: effects of ageing. AB - We have examined the ability of 1,25(OH)2-vitamin D3 [1,25(OH)2D3; calcitriol], the hormonal form of vitamin D3, to stimulate the phosphorylation of proteins in rat duodenum from young (3 months) and aged (22-24 months) rats. Brief (30 s) exposure of duodenum preincubated with 32P-orthophosphate to the hormone increased the labeling of whole tissue proteins, an effect that was greatly diminished in aged animals. The response was dose-dependent, with maximal stimulation achieved at 1 nM calcitriol (+113% and +10% for young and aged rats, respectively). Phosphoproteins were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and identified by autoradiography. The hormone potentiated the phosphorylation predominantly on serine, threonine, and tyrosine residues of five acidic proteins of relative molecular masses of 66, 48, 45, 28, and 16 kDa. Moreover, the effects of calcitriol were exerted at the membrane level and varied as a function of exposure time. Direct treatment of purified basal lateral membranes for 30 s with the hormone (1 nM) stimulated the incorporation of 32P of a 66 kDa protein by 75% and of a 48 and 45 kDa proteins by 60%. The effects of the hormone on basal lateral membrane protein phosphorylation were suppressed by the PKA, PKC, and tyrosine kinase inhibitors, Rp-cAMPS, bisindolylmaleimide, and genistein, respectively. In basal lateral membrane isolated from old animals, only minor changes in calcitriol-induced protein phosphorylation of the 66-kDa protein were observed. Taken together, these results suggest that calcitriol modulates duodenal membrane protein phosphorylation, at least in part through PKA, PKC, and tyrosine kinases, and that this mechanism is severely altered with ageing. The identity of the proteins whose phosphorylation was stimulated by calcitriol and their physiological role is currently under investigation. PMID- 10673152 TI - Investigation of the endocrine system in extended longevity lines of Drosophila melanogaster. AB - There is a complete absence of information about the endocrinology of Drosophila melanogaster in relation to genetic-based differential longevity in this model species. In the present study, aspects of the endocrine system of D. melanogaster were investigated in selected and control lines characterized by relative differences in life span. By using extracts from whole bodies, steroid hormone (ecdysteroid) titers were determined by radioimmunoassay in all replicate selected and control lines on the first and fourth day of adult life. The results suggest that ecdysteroid titers were relatively reduced on the first day post eclosion in females from the long-lived lines, but this difference was not present on the fourth day posteclosion. The reduction in early-age ecdysteroid titers in long-lived females might be related to the decrease in early-age fecundity in the selected lines. There was no difference between line types in male ecdysteroid titers on either day post-eclosion. Two classes of enzymes that act on juvenile hormone were also investigated in the present study. Esterase and epoxide hydrolase activity on juvenile hormone was assessed in females in all replicate selected and control lines at approximately 12 h or 4 days post eclosion. There was no difference between selected and control lines in the specific activity of either class of enzymes that metabolize juvenile hormone. PMID- 10673153 TI - Age-dependent hypertension in Mpv17-deficient mice, a transgenic model of glomerulosclerosis and inner ear disease. AB - The mutant mouse strain Mpv17-/-, carries a retroviral germline integration that inactivates the Mpv17 gene. Mpv17-deficient mice develop progressive glomerulosclerosis and sensineural deafness at early age. Characteristic basement membrane alterations are found in both sites of pathology. Mpv17 is a peroxisomal protein involved in the metabolism of reactive oxygen species, yet its molecular function is unknown. Dysregulation of antioxidant enzymes and basal membrane components has been established in this model and successful therapeutic intervention with antioxidants prove the causal role of reactive oxygen species in the development of the disease phenotype. We here investigated if the Mpv17-/- mice might be hypertensive. Indeed, our study revealed that Mpv17-/- mice developed significant systemic hypertension and tachycardia between 4 weeks and 5 months of age, accompanied by polyuria and elevated natriuresis. Judging from serum and urine parameters, the hypertensive condition develops concomitantly with the renal disease. Biochemical and pharmacological studies that used the endothelin receptor antagonist bosentan and the angiotensin converting enzyme inhibitor cilazapril indicated no involvement of the endothelin and renin angiotensin systems in this hypertension, suggesting a potential novel mechanism of blood pressure regulation in this new murine hypertension model. Thus, Mpv17-/ mice unravel an intriguing new association between a defect in reactive oxygen metabolism and the age-dependent development of hypertension. PMID- 10673154 TI - Characterization of four distinct monoclonal antibodies specific to BmK AS-1, a novel scorpion bioactive polypeptide. AB - Four monoclonal antibodies designed as 2#, 3#, 4# and 5# have been raised against a novel bioactive polypeptide BmK AS-1 purified from the Chinese scorpion Buthus martensi Karsch. All of these antibodies exhibited specific affinity with antigen by ELISA and Biosensor assay. Western blot analysis showed that 3# and 4# were able to recognize the denatured antigen, but not 2# and 5#. These antibodies could cross-react with BmK AS, but not with other types of BmK neurotoxins such as BmK I (an alpha-like toxin) and BmK IT (an excitatory insect-selective toxin), and in which only 5# can partially react with BmK IT2 (a depressant insect selective toxin). Immunocytochemical staining demonstrated that 3#, 4# and 5# antibodies can visualize the antigen bound to the membrane of SK-N-SH neuroblast cells, with the exception of 2#. This suggests that either conformation alteration of receptor binding might be prone to nonvisualization or the epitope recognized by antibody 2# might be overlapped with receptor binding sites of antigen. The antibodies developed in the study should provide powerful new tools for investigating the structure/function relationship and pharmacological mechanism of scorpion neurotoxins. PMID- 10673155 TI - Monitoring of microcystin-protein phosphatase adduct formation with immunochemical methods. AB - Using anti-microcystin-LR monoclonal antibodies, an immunoblotting procedure was developed to monitor the formation of microcystin-protein phosphatase adducts in vitro and in vivo. The detection limits for the covalent binding of MCYST-LR with the recombinant protein phosphatase 1 (PP1) and rabbit liver cytosol proteins were found to be 0.1 ng and 0.3 ng per assay, respectively. MCYST-PP1 adducts were detected 30 s after the addition of MCYST-LR into the reaction mixture. Reduction of the methyldehydroalanine (Mdha) residue of MCYST-LR with ethanethiol totally abolished the covalent binding of the toxin to PP1, but retained its inhibitory toxicity on PP1. Immunoblotting analyses and enzyme-linked immunosorbant assay showed that between 5 min to 16 h after i.p. injection of single dose (35 microg/kg) of MCYST-LR into mice, approximately 0-27% of the injected toxin was found covalently bound while 0.2-9.2% existed as free form in liver cytosol. PMID- 10673156 TI - Horse IgG isotypes and cross-neutralization of two snake antivenoms produced in Brazil and Costa Rica. AB - Horse IgG isotypes and cross-neutralization of two snake antivenoms produced in Brazil and Costa Rica. Toxicon 000-000. This work compared the specificity, ELISA titers and IgG subclass content of the polyvalent antivenom (anti-Bothrops asper, Crotalus durissus durissus and Lachesis muta stenophrys) of Instituto Clodomiro Picado (Costa Rica) and the bothropic antivenom (anti-Bothrops jararaca, B. jararacussu, B. moojeni, B. neuwiedi and B. alternatus) of Instituto Butantan (Brazil). The role of IgG(T) and IgGa subclasses in neutralization of some venom toxic activities and the cross neutralization of the antivenoms against B. jararaca and B. asper venoms were also evaluated. Both antivenoms were able to recognize B. asper and B. jararaca venoms by immunoblotting and presented similar antibody titers when assayed by ELISA. IgG(T) was highest, followed by IgGa, IgGb and IgGc. IgGa and IgG(T) isotypes isolated from both antivenoms by affinity chromatography were tested for neutralization of lethal, hemorrhagic, coagulant and phospholipase A2 activities of the homologous venoms. In both antivenoms, IgG(T) was the major isotype responsible for neutralization of all the tested activities, followed by IgGa. These results suggest that Instituto Butantan and Instituto Clodomiro Picado antivenoms have the same IgG profile and their neutralizing ability is due mostly to the IgG(T) isotype. Also, they neutralize lethality in mice induced by homologous and heterologous venoms, the bothropic antivenom of Instituto Butantan being more effective. PMID- 10673157 TI - The isolation and characterization of a peptide that alters sodium channels from Buthus martensii Karsch. AB - The peptides were purified using gel filtration, ion exchange, FPLC, and HPLC chromatography and found to greatly prolong action potentials at nanomolar concentrations when applied to frog and mouse nerves. The N-terminal primary amino acid sequence of one of the peptides, BMK 16(5), was determined. The first 23 amino acids of BMK 16(5) were found to be: VKDGYIADDRNCPYFCGRNAYYD. The two cysteine residues in the sequence appeared as Edman sequence cycle blanks; however, they were assigned to be cysteines due to sequence similarity to other peptide toxins that bind to sodium channels and identification of the presence of cysteines obtained from single time point amino acid analysis. The MW of BMK 16(5) was determined by a Perkin Elmer API 300 LC/MS/MS to be 3,695. The amino acid residues of BMK 16(5) show strong similarity with the first 23 amino acid residues of a number of scorpion alpha neurotoxins. Unlike these neurotoxins, BMK 16(5) possesses a proline residue at position 13 which will likely make it fold in a unique way so as to bind to and alter sodium channels. PMID- 10673158 TI - The gene cloning and sequencing of Bm-12, a chlorotoxin-like peptide from the scorpion Buthus martensi Karsch. AB - According to the known amino acid sequence of Bm-12, a short chain insect neurotoxin from the venom of the scorpion Buthus martensi Karsch (BmK) with considerable primary sequence homology to chlorotoxin, the gene specific primers were designed and synthesized for 3' and 5'RACE (Rapid Amplification of cDNA Ends). The two partial cDNA fragments obtained by 3' and 5'RACE were cloned and sequenced, and the full length cDNA sequence of Bm-12 was then completed by overlapping these two partial cDNA sequences. The predicted amino acid sequence consists of 59 amino acid residues including a putative signal peptide of 24 residues and a mature toxin of 35 residues. The predicted amino acid sequence of Bm-12 was almost consistent with the determined, different only in one residue at position 27, Lys was replaced by Gly. Based on the determined cDNA sequence, and using the total DNA isolated from the scorpion venom glands as a template, the genomic DNA of Bm-12 was also amplified by PCR and sequenced. The genomic DNA sequence revealed an intron of 93 bp present within the signal peptide region. PMID- 10673159 TI - Characterization of mice antisera elicited with a ciguatoxin tetracyclic synthetic ring fragment (JKLM) conjugated to carrier proteins. AB - As a good alternative to the lack of pure ciguatoxin (CTX), conjugates of JKLM ring fragment, a carboxylic derivative of the right-hand tetracyclic terminus portion of CTX-1B (the most potent CTX) with two carrier proteins have been synthesized. Two procedures using different amount of hapten were evaluated: (i) a bulk technique (3-5 mg) via the N-hydroxysuccinimide ester of the carboxylic fragment in the presence of a water-soluble carbodiimide according to the standard method in aqueous buffer, or (ii) a micro-scale technique (300 microg) via the mixed anhydride method performed in a reversed micellar medium. In both cases, bovine serum albumin and ovalbumin were respectively used for immunization of BALB/c mice and antibody screening by a solid phase enzyme-linked immunosorbent assay (ELISA). Using the conjugates obtained through the micro scale procedure, a long-term immunization schedule appeared to be more efficient to specifically trigger the mice immune system. These antisera titers determined in an end-point titration standard ELISA format were found around 1/128,000 as compared to 1/16,000 obtained in the short-term protocol (immunogen prepared via the bulk procedure). In competitive inhibition ELISA experiments, both types of antisera did not significantly cross-react with a brevetoxin congener (PbTx-3), okadaic acid (OA), monensin or other polyether compounds, but only sera from the short-term protocol did show high cross-reactivity to CTX-1B (133%). With sera from the long-term protocol, a lower detection limit for JKLM (1.23 x 10(-9) M) was achieved by implementation of a biotin-avidin amplification system rather than by miniaturization of the assay in Terasaki plates. This study confirms the feasibility of the immunological approach for CTXs assay in fish tissues, but also emphasizes the importance of (i) the choice of the hapten to construct a relevant well-defined immunogen, (ii) the immunization schedule to obtain hapten specific Abs still exhibiting high cross-reactivity to CTXs. PMID- 10673160 TI - Sea snake Hydrophis cyanocinctus venom. II. Histopathological changes, induced by a myotoxic phospholipase A2 (PLA2-H1). AB - A toxic phospholipase A2 (PLA2-H1), isolated from the venom of the sea snake Hydrophis cyanocinctus, was tested for its ability to induce myonecrosis and histopathological changes in albino rats and mice. Induction of myonecrosis was demonstrated by their ability to release creatine kinase (CK) from damaged muscle fibers and direct histopathological examination of the injected muscles (i.m.). PLA2-H1 exhibits intense myonecrosis characterized by the changes including, necrosis and edematous appearance with cellular infiltrate, vacuolation and degenerated muscle cells with delta lesions and heavy edema in between the cells. No myoglobinuria was noted in any group of animals. The purified PLA2-H1 was also administered intraperitoneally into the experimental animals and tissue samples were taken at several time intervals. Light microscopic examination of the kidney sections revealed severe damage, evident by focal tubular necrosis, complete disquamation of epithelial lining and epithelial degeneration of tubules in all test animals. Light micrographs of liver sections after 24 h of injection shows fatty infiltration in parenchyma and squashed hepatocytes, while after 48 h, fatty vacuolation of parenchyma in a generalized pattern was observed. Furthermore, sections of the lungs of the same group of animals (48 h) show dilated bronchia and marked infiltration of inflammatory cells within alveoli. Our results suggest that the purified PLA2-H1 induced moderate myotoxicity in muscles and mild histopathological changes in other vital organs without myoglobinuria. PMID- 10673161 TI - Amino acid sequence and biological properties of the lectin from the venom of Trimeresurus okinavensis (Himehabu). AB - A lectin was isolated from the venom of Trimeresurus okinavensis (Himehabu) and the complete amino acid sequence was determined using clostripain, lysyl endopeptidase, and V8 protease. The hemagglutinating activity of this lectin are reported. PMID- 10673162 TI - California ground squirrel (Spermophilus beecheyi) blood sera inhibits crotalid venom proteolytic activity. AB - Some California ground squirrels (Spermophilus beecheyi) show limited necrosis following envenomation by northern Pacific rattlesnakes (Crotalus viridis oreganus). This study demonstrates that S. beecheyi blood sera inhibits venom proteases. Sera from rattlesnake-abundant habitats inhibited C. v. oreganus venom more effectively than venom from two allopatric rattlesnake species, C. v. viridis and C. atrox, suggesting evolutionary specialization. The pattern of inhibition among squirrel populations corresponds best with history of rattlesnake predation, in contrast to current rattlesnake density. PMID- 10673163 TI - Fang tip spread, puncture distance, and suction for snake bite. AB - We measured the distance between fang tip punctures in defensive bites by western diamondback rattlesnakes (Crotalus atrox) and the distance between their retracted fangs. Because the fang tips at penetration average 112% further apart than their bases at rest, The Extractor, a device widely marketed in the United States for snake bite first aid, will not simultaneously cover both punctures of most adult New World pitvipers. PMID- 10673164 TI - Toxicities and distribution of tetrodotoxin in the tissues of puffer fish found in the coast of the Baja California Peninsula, Mexico. AB - Toxicities and tetrodotoxin distribution in tissues of five puffer fish species commonly found in the littoral of Baja California Peninsula, Mexico (Sphoeroides annulatus, S. lobatus, S. lispus, Arothron meleagris and Canthigaster punctatissima) were evaluated by bioassay and HPLC. The toxicities estimated as tetrodotoxin-equivalents of all species were more than 0.42 microg/g in at least one of the tissues tested, and the highest was found in S. lispus liver (130 microg/g). PMID- 10673165 TI - Inhaled corticosteroids are beneficial in chronic obstructive pulmonary disease. PMID- 10673168 TI - Rebuttal from dr. barnes PMID- 10673167 TI - Rebuttal from dr. calverley PMID- 10673166 TI - Inhaled corticosteroids are not beneficial in chronic obstructive pulmonary disease. PMID- 10673169 TI - My initiation into respiratory physiology. PMID- 10673170 TI - Upper airway collapsibility and cephalometric variables in patients with obstructive sleep apnea. AB - Increased pharyngeal collapsibility and abnormal anatomic structures have been postulated to contribute to the pathophysiology of obstructive sleep apnea (OSA) syndrome. It is unclear whether the abnormal craniofacial and soft tissue features may affect the pharyngeal collapsibility and contribute to the apnea density. In the present study we examine the relationship between pharyngeal collapsibility and cephalometric variables in a group of 57 male OSA patients. Pharyngeal collapsibility was measured during the night of nasal continuous positive airway pressure (nCPAP) titration by analyzing the pressure-flow relationship. Pharyngeal critical pressure (Pcrit) was calculated as the extrapolated pressure at zero flow. The patients, age 52.0 +/- 9.0 yr, had an average apnea-hypopnea index (AHI) of 72.6 +/- 31.8 and a mean Pcrit of 2.4 +/- 1.0 cm H(2)O. A significant correlation was found between Pcrit and the soft palate length (SPl) (r = 0.27, p = 0.04), the distance from the hyoid bone to the posterior pharyngeal wall (H-Ph) (r = 0. 29, p = 0.03), and the distance from the hyoid bone to posterior nasal space (H-Pns) (r = 0.32, p = 0.02). While in obese patients Pcrit was related to SPl and neck circumference, the distance of the hyoid bone to the mandibular plane (H-MP) affected Pcrit in nonobese patients. Our results show that both pharyngeal soft tissue abnormalities and the lower position of the hyoid bone affect Pcrit in OSA patients, suggesting that an anatomic narrowing contributes to the upper airway collapsibility. PMID- 10673171 TI - Forced expiratory flows and volumes in infants. Normative data and lung growth. AB - Forced expiratory flows (FEF) can be measured in infants from lung volumes initiated near total lung capacity. In order to establish reference values and to evaluate lung growth, we obtained measurements in 155 healthy subjects between 3 and 149 wk of age. Forced vital capacity (FVC) was highly correlated with body length; however, after accounting for length, age was also significant. When subjects were divided at the median age (40 wk) younger compared with older subjects had a significantly larger slope for length (3.7 versus 2.8; p = 0.002). The flow parameters (FEF(50), FEF(75), FEF(85), and FEF(25-75)) were highly correlated with length, and those infants whose mothers smoked had lower flows. For FEF(75), male subjects had lower flows than female subjects. The relationship between FEF and volume was assessed using FEV(0.5)/FVC, which decreased with increasing length. Smaller subjects emptied their lung volume proportionately faster. We conclude that our study provides reference values for this age group and demonstrates that smoke-exposed infants and male subjects have decreased FEF. In addition, our findings indicate that lung volume increases most rapidly during the first year of life and that airways are large relative to lung volume very early in life. PMID- 10673172 TI - Effect of the prone position on patients with hydrostatic pulmonary edema compared with patients with acute respiratory distress syndrome and pulmonary fibrosis. AB - This study examined the effect of the prone position on mechanically ventilated patients with hydrostatic pulmonary edema (HPE). Eight patients with acute HPE and mechanically ventilated in the prone position (Group 1) were studied. Six patients with acute HPE and mechanically ventilated in the supine position (Group 2), 20 patients with ARDS (Group 3), and 5 patients with pulmonary fibrosis (PF) (Group 4) served as control patients. Patients with HPE, who after being mechanically ventilated for at least 6 h needed an FI(O(2)) >/= 0.6 to achieve an Sa(O(2)) of approximately 90%, and did not respond to recruitment maneuvers, were turned to the prone position. Parameters of oxygenation, lung mechanics, and hemodynamics were determined in both the supine and prone positions. All patients with HPE exhibited improvement of oxygenation when they were placed in the prone position. The Pa(O(2))/FI(O(2)) ratio increased from 72 +/- 16 in the supine position to 208 +/- 61 after 6 h in the prone position (p < 0.001); the rise in Pa(O(2)) was persistent, without detrimental effect on hemodynamics. Fifteen of 20 patients with ARDS (75%) improved oxygenation when in the prone position. The Pa(O(2))/FI(O(2)) ratio increased from 83 +/- 14 in the supine position to 189 +/ 34 after 6 h in the prone position (p < 0.001). In contrast, 5 of 20 patients with ARDS (25%) and none of the patients with PF responded favorably to prone positioning. Patients with HPE and early ARDS responded better to prone positioning than did patients with late ARDS and PF. Patients with HPE and ventilated in the supine position had a lower Pa(O(2))/FI(O(2)) ratio and the duration of mechanical ventilation was longer compared with that of patients in the prone position. Our results show that the prone position may be a useful maneuver in treating patients with severe hypoxemia due to pulmonary edema. The presence of pulmonary edema, as in early ARDS and HPE predicts a beneficial effect of the prone position on gas exchange. In contrast, the presence of fibrosis, as in late ARDS and pulmonary fibrosis, predisposes to nonresponsiveness to prone positioning. PMID- 10673173 TI - Effects of varying approaches for identifying respiratory disturbances on sleep apnea assessment. AB - Varying approaches to measuring the respiratory disturbance index (RDI) may lead to discrepant estimates of the severity of sleep-disordered breathing (SDB). In this study, we assessed the impact of varying the use of corroborative data (presence and degree of desaturation and/or arousal) to identify hypopneas and apneas. The relationships among 10 RDIs defined by various definitions of apneas and hypopneas were assessed in 5,046 participants in the Sleep Heart Health Study (SHHS) who underwent overnight unattended 12-channel polysomnography (PSG). The magnitude of the median RDI varied 10-fold (i.e., 29.3 when the RDI was based on events identified on the basis of flow or volume amplitude criteria alone to 2.0 for an RDI that required an associated 5% desaturation with events). The correlation between RDIs based on different definitions ranged from 0.99 to 0.68. The highest correlations were among RDIs that required apneas and hypopneas to be associated with some level of desaturation. Lower correlations were observed between RDIs that required desaturation as compared with RDIs defined on the basis of amplitude criteria alone or associated arousal. These data suggest that different approaches for measuring the RDI may contribute to substantial variability in identification and classification of the disorder. PMID- 10673174 TI - Time course of sleep-related breathing disorders in first-ever stroke or transient ischemic attack. AB - To investigate the prevalence and behavior of sleep-related breathing disorders (SRBDs) associated with a first-ever stroke or transient ischemic attack (TIA), we prospectively studied 161 consecutive patients admitted to our stroke unit. Complete neurological assessment was performed to determine parenchymatous and vascular localization of the neurological lesion. Stroke subtype was categorized as TIA, ischemic (IS), or hemorrhagic (HS). A portable respiratory recording (PRR) study was performed within 48-72 h after admission (acute phase), and subsequently after 3 mo (stable phase). During the acute phase, 116 patients (71.4%) had an apnea-hypopnea index (AHI) > 10 events/h and 45 (28%) had an AHI > 30. No relationships were found between sleep-related respiratory events and the topographical parenchymatous location of the neurological lesion or vascular involvement. Cheyne-Stokes breathing (CSB) was observed in 42 cases (26.1%). There were no significant differences in SRBD according to the stroke subtype except for the central apnea index (CAI). During the stable phase a second PRR was performed in 86 patients: 53 of 86 had an AHI > 10 and 17 of 86 had an AHI > 30. The AHI and CAI were significantly lower than those in the acute phase (16.9 +/- 13.8 versus 22.4 +/- 17.3 and 3.3 +/- 7.6 versus 6.2 +/- 10.2, respectively) (p < 0.05) while the obstructive apnea index (OAI) remained unchanged. CSB was observed in 6 of 86 patients. The prevalence of SRBD in patients with first-ever stroke or TIA is higher than expected from the available epidemiological data in our country. No correlation was found between neurological location and the presence or type of SRBD. Obstructive events seem to be a condition prior to the neurological disease whereas central events and CSB could be its consequence. PMID- 10673176 TI - Differential metabolism of arachidonic acid in nasal polyp epithelial cells cultured from aspirin-sensitive and aspirin-tolerant patients. AB - The mechanism of aspirin (acetylsalicylic acid [ASA]) sensitivity associated with severe asthma and chronic rhinosinusitis with nasal polyps ("aspirin triad") has been attributed to arachidonic metabolism alternations, although the putative biochemical defects have not been elucidated. The aim of this study was assessment of the hypothesis that local production of eicosanoids in the respiratory epithelium in patients with ASA-sensitive asthma/rhinosinusitis (ASARS) differs from that of ASA-tolerant patients with rhinosinusitis (ATRS). Nasal polyps were obtained from 10 patients with ASARS and 15 with ATRS during routine polypectomies, and epithelial cells (ECs) were cultured on bovine collagen type I matrix (Vitrogen 100), in medium supplemented with growth factors. The generation of eicosanoids in supernatants of confluent ECs (6 to 8 d of culture; purity > 98%) was quantified by immunoassays. Unstimulated ECs from ASARS patients generated significantly less prostaglandin E(2)(PGE(2)) compared with ATRS (0.8 +/- 0.3 versus 2. 4 +/- 0.5 ng/microg double-stranded deoxyribonucleic acid [dsDNA], respectively), although a similar relative increase in response to calcium ionophore and inhibition with ASA was observed in both groups. Basal levels of 15-hydroxyeicosatetraenoic acid (15-HETE) were not different between groups, and calcium ionophore enhanced its production to a similar extent. However, cells incubation with 200 microM ASA for 60 min resulted in a significant increase (mean +359%) in 15-HETE generation only in ASARS patients, whereas no effect of ASA on 15-HETE generation in ATRS patients was observed. PGF(2alpha) generation was similar in both groups, and no significant generation of PGD(2) or leukotriene C(4) (LTC(4)) was observed in epithelial cell cultures from either group. Our results indicate that nasal polyps ECs from ASA sensitive patients have significant abnormality in basal and ASA-induced generation of eicosanoids which may be causally related to the mechanism of ASA sensitivity. PMID- 10673177 TI - Activation of neutrophils, eosinophils, and lymphocytes in the lower respiratory tract in Wegener's granulomatosis. AB - Levels of cell products released by neutrophils, eosinophils and lymphocytes were measured in the bronchoalveolar lavage fluid (BALF) of 19 patients with pulmonary active Wegener's granulomatosis (WG) to assess in vivo the magnitude of cellular activation at sites of active disease. Measurements included the BAL cell profile and BALF levels of myeloperoxidase (MPO), free proteinase 3 (fPR3), complexes of PR3 and alpha1-antitrypsin (PR3/alpha1-AT), eosinophil cationic protein (ECP), peroxidase activity (PEROX), and soluble interleukin-2 receptor (sIL-2R). Six patients also underwent a repeat examination after immunosuppressive treatment. Pulmonary active WG was found to be associated with elevated MPO, PEROX, ECP, and sIL-2R levels in BALF. Only trace amounts of fPR3 were detected, the bulk of PR3 being found in PR3/alpha1-AT complexes. Clinically effective treatment depressed BAL neutrophil counts and reversed elevated levels of MPO and PEROX but had an inconsistent effect on the BAL lymphocyte count and the sIL-2R level. In conclusion, the elevated levels of extracellular MPO and PEROX at a site of active disease and the correlation between these and clinical disease activity support the view that neutrophils are indeed an important effector cell population in WG lung disease. The present data also suggest that oxidative injury is an important aspect of neutrophil-mediated lung injury, whereas it remains unresolved whether the low levels of fPR3 in the BALF adequately reflect the situation at inflammatory tissue sites. PMID- 10673175 TI - Smoking cessation and lung function in mild-to-moderate chronic obstructive pulmonary disease. The Lung Health Study. AB - Previous studies of lung function in relation to smoking cessation have not adequately quantified the long-term benefit of smoking cessation, nor established the predictive value of characteristics such as airway hyperresponsiveness. In a prospective randomized clinical trial at 10 North American medical centers, we studied 3, 926 smokers with mild-to-moderate airway obstruction (3,818 with analyzable results; mean age at entry, 48.5 yr; 36% women) randomized to one of two smoking cessation groups or to a nonintervention group. We measured lung function annually for 5 yr. Participants who stopped smoking experienced an improvement in FEV(1) in the year after quitting (an average of 47 ml or 2%). The subsequent rate of decline in FEV(1) among sustained quitters was half the rate among continuing smokers, 31 +/- 48 versus 62 +/- 55 ml (mean +/- SD), comparable to that of never-smokers. Predictors of change in lung function included responsiveness to beta-agonist, baseline FEV(1), methacholine reactivity, age, sex, race, and baseline smoking rate. Respiratory symptoms were not predictive of changes in lung function. Smokers with airflow obstruction benefit from quitting despite previous heavy smoking, advanced age, poor baseline lung function, or airway hyperresponsiveness. PMID- 10673178 TI - Long-term follow-up of pulmonary function in patients with nasal polyposis. AB - The outcome of asthma and/or nonspecific bronchial hyperresponsiveness (BHR) associated with nasal polyposis (NP) is uncertain. Over a 4-yr period, we investigated the long-term changes of pulmonary function and BHR in 46 patients with NP. Each subject was assessed for nasal symptoms and tested for allergy skin prick tests, serum total IgE, spirometry, and carbachol challenge at baseline before initiating any treatment (T0). Nasal symptoms evaluation, spirometric measurements, and carbachol challenge were repeated at T1 and at T2 (respectively, 12.7 +/- 0.9 and 47.9 +/- 2. 2 mo after T0). In addition, bronchodilator response was measured at T2. At T0, 25 patients exhibited BHR and 16 of 25 were asthmatic. All patients were treated first with topical steroids for 6 wk (beclomethasone 600 microg/d). Eighteen patients were successfully treated with topical steroids (topical steroids responders). Intranasal ethmoidectomy was performed in 28 patients who did not improve with topical steroids alone (topical steroids nonresponders). Nasal score improved at T1 and remained improved at T2 as compared with T0 in both groups (p < 0.005). Topical steroids nonresponders demonstrated a significant decrease of FEV(1), FEV(1)/FVC ratio, and FEF(25-75) at T1 (p < 0.05) and at T2 (p < 0.0005), whereas no significant change was observed in FEV(1) and FEV(1)/FVC ratio in responders. DeltaFEV(1) (%) between T2 and T0 was not related to the presence of asthma, BHR, or atopy. Bronchodilator response at T2 was similar in the two groups. BHR did not significantly change over the 4-yr follow-up period in the two groups. No change in pulmonary symptoms and/or asthma severity occurred. Our results show that nonreversible airflow obstruction appears over a 4-yr follow-up period in topical steroids nonresponders patients with NP requiring nasal surgery. The long term contribution of these changes to the development of respiratory symptoms in patients with NP remains to be documented. PMID- 10673179 TI - The effects of chronic alcohol abuse on pulmonary glutathione homeostasis. AB - The incidence and severity of the acute respiratory distress syndrome (ARDS) is increased in critically ill patients with a prior history of chronic alcohol abuse; however, the specific mechanisms responsible for this association are unknown. Recently, we determined that chronic ethanol ingestion in rats decreased the alveolar epithelial lining fluid (ELF) concentration of the antioxidant glutathione (GSH), which is a characteristic finding in patients with ARDS. However, the effects of chronic alcohol abuse on the human alveolar epithelium are essentially unknown. Therefore, as a first step we asked if chronic alcohol abuse, independent of other comorbid conditions, decreases the concentration of GSH in the human lung. We determined that otherwise healthy chronic alcoholics had significantly decreased ELF concentrations of GSH compared with nonalcoholic control subjects (79 micromol [48 to 118 micromol] versus 576 micromol [493 to 728 mmol], p < 0.001). Furthermore, the percentage of GSH in the oxidized form was higher in the chronic alcoholics (9.8% [2.2 to 14.8%] versus 2.8% [0.4 to 4.0%] p = 0.05), indicative of increased utilization of GSH. This is the first report that chronic alcohol abuse alters GSH homeostasis in the human lung, and suggests a potential mechanism by which chronic alcohol abuse predisposes susceptible patients to develop ARDS. PMID- 10673180 TI - An oral elastic mandibular advancement device for obstructive sleep apnea. AB - Oral mandibular advancement devices are becoming an increasingly important treatment alternative for obstructive sleep apnea (OSA). The first aim of the study was to determine whether a new oral elastic mandibular advancement device (EMA) prevents pharyngeal airway closure during sleep in patients with OSA. The second aim of the study was to determine if the polysomnographic response to the oral mandibular advancement device was dependent on the site of airway closure. Overnight polysomnograms were performed in 28 untreated OSA subjects with and without EMA. A third polysomnogram was performed in 12 of the subjects to determine the site of airway closure without the device. Site of airway closure above or below the oropharynx was determined by measuring the respective presence or absence of respiratory fluctuations in oropharyngeal pressure during induced occlusions in non-rapid eye movement (NREM) sleep. Mean apnea-hypopnea index (AHI) was 52.6 +/- 28.2 (SD) events/h without the device and 21.2 +/- 19.3 events/h with the device. Nineteen subjects (68%) had at least a 50% reduction in AHI with the device. The change in AHI with the device (AHI without device - AHI with device) was directly related to the AHI without the device. All three subjects with airway closure in the lower pharyngeal airway had a greater than 80% reduction in AHI with the device. Two of the nine subjects with airway closure in the velopharynx had a similar therapeutic response. The results show the effectiveness of EMA in the treatment of OSA. The results also indicate that polysomnographic severity of OSA and the site of airway closure should not be used to exclude patients from this oral device treatment. PMID- 10673181 TI - Sleep onset REM periods during multiple sleep latency tests in patients evaluated for sleep apnea. AB - Although 2 or more sleep onset rapid eye movement (REM) periods (2omSOREMPs) on a Multiple Sleep Latency Test (MSLT) raise the possibility of narcolepsy, patients with obstructive sleep apnea (OSA) also can have 2omSOREMPs, which may then cause diagnostic uncertainty. To explore what features among OSA patients predict 2omSOREMPs on an MSLT that follows nocturnal polysomnography, we reviewed data from 1,145 consecutively studied patients suspected or confirmed to have OSA rather than narcolepsy. Overall, 4.7% of the subjects had 2omSOREMPs. Variables that were independently predictive of 2omSOREMPs in logistic regression models included male gender (OR = 4.4, 95% CI = 1.9 to 12.7), a 5-min decrease in the MSLT-derived mean sleep latency (OR = 1.9, 95% CI = 1.3 to 2.8), a 90-min decrease in nocturnal latency to REM sleep (OR = 1.6, 95% CI = 1.1 to 2.5), and a 15-unit decrease in minimal recorded oxygen saturation (OR = 1.6, 95% CI = 1.3 to 2.0). We conclude that among patients suspected or confirmed to have OSA, one or more of these four variables-male sex, sleepiness, nocturnal REM sleep latency, and extent of oxygen desaturation-could reflect neurophysiological mechanisms responsible for 2omSOREMPs. Consideration of these variables, and especially gender, may be useful in clinical practice when 2omSOREMPs are found unexpectedly. PMID- 10673182 TI - An objective analysis of the pressure-volume curve in the acute respiratory distress syndrome. AB - To assess the interobserver and intraobserver variability in the clinical evaluation of the quasi-static pressure-volume (P-V) curve, we analyzed 24 sets of inflation and deflation P-V curves obtained from patients with ARDS. We used a recently described sigmoidal equation to curve-fit the P-V data sets and objectively define the point of maximum compliance increase of the inflation limb (P(mci, i)) and the true inflection point of the deflation limb (P(inf,d)). These points were compared with graphic determinations of lower Pflex by seven clinicians. The graphic and curve-fitting methods were also compared for their ability to reproduce the same parameter value in data sets with reduced number of data points. The sigmoidal equation fit the P-V data with great accuracy (R(2) = 0.9992). The average of Pflex determinations was found to be correlated with P(mci,i) (R = 0.89) and P(inf,d) (R = 0.76). Individual determinations of Pflex were less correlated with the corresponding objective parameters (R = 0.67 and 0.62, respectively). Pflex + 2 cm H(2)O was a more accurate estimator of P(inf,d) (2 SD = +/-6.05 cm H(2)O) than Pflex was of P(mci,i) (2 SD = +/-8.02 cm H(2)O). There was significant interobserver variability in Pflex, with a maximum difference of 11 cm H(2)O for the same patient (SD = 1.9 cm H(2)O). Clinicians had difficulty reproducing Pflex in smaller data sets with differences as great as 17 cm H(2)O (SD = 2.8 cm H(2)O). In contrast, the curve-fitting method reproduced P(mci,i) with great accuracy in reduced data sets (maximum difference of 1.5 cm H(2)O and SD = 0.3 cm H(2)O). We conclude that Pflex rarely coincided with the point of maximum compliance increase defined by a sigmoid curve-fit with large differences in Pflex seen both among and within observers. Calculating objective parameters such as P(mci,i) or P(inf,d) from curve-fitted P-V data can minimize this large variability. PMID- 10673183 TI - Effects of isoproterenol on diaphragmatic contractility in septic peritonitis. AB - We investigated the effects, and the mechanism of the effects, of isoproterenol on diaphragmatic contractility and fatigue in septic peritonitis in vitro. Ninety six rats were divided into two groups of 48. One group (CLP group) was treated with cecal ligation and perforation (CLP) and the other (sham group) was treated with laparotomy. The left hemidiaphragm was removed at 16 h after the operation. We assessed the diaphragmatic contractility by twitch characteristics and force frequency curves in vitro. Diaphragm fatigue was induced by rhythmically stimulating strips to contract at 60/ min (20 Hz, 0.33-s trains, 1 train/s) over a 4-min period. Force-frequency curves were determined before and after fatigue. Isoproterenol (10(-9), 10(-8), and 10(-7) M), a beta-adrenoceptor agonist, was cumulatively administered to the organ bath. Isoproterenol significantly increased diaphragmatic contractility. There were no significant changes in diaphragmatic contractility in the sham group. Isoproterenol (10(-7) M) significantly accelerated diaphragmatic recovery of fatigue and increased cAMP levels both in the sham group and the CLP group. Propranolol (10(-7) M), a general beta-adrenoceptor blocker, completely abolished the positive inotropic effect of isoproterenol (10(-7) M) and increased cAMP levels in the CLP group. Dibutyryl cAMP (10(-3) M), a derivative of cyclic AMP, mimicked the effects of isoproterenol in the CLP group. These results suggest that isoproterenol increases diaphragmatic contractility and accelerates diaphragmatic recovery of fatigue in septic peritonitis by activating the adenylate cyclase system. PMID- 10673184 TI - Quadriceps fatigue after cycle exercise in patients with chronic obstructive pulmonary disease. AB - Patients with COPD have derangements in respiratory mechanics that may cause them to stop exercising before the exercising limb muscles reach their functional limits. However, because lung disease makes activity unpleasant, patients with chronic obstructive pulmonary disease (COPD) often adapt a sedentary lifestyle leading to progressive deconditioning. Deconditioning will lead to progressive deterioration in limb muscle function, which could adversely affect exercise capacity. The purpose of this study was to determine whether fatigue of the quadriceps muscle occurs after high intensity cycle exercise to the limits of tolerance in patients with moderate to severe COPD. Nineteen male patients with COPD (FEV(1) 1.54 +/- 0. 12 L; 42 +/- 3% predicted) exercised at 60 to 70% of their predetermined maximal work capacity until exhaustion. The femoral nerve was supramaximally stimulated with a figure-of-eight magnetic coil, and quadriceps twitch force (TwQ) was measured before and at 10, 30, and 60 min postexercise. Patients exercised at 53.7 +/- 4.1 watts for 10.4 +/- 1.4 min. Peak V O(2) was 1.24 +/- 0.08 L/min (51. 3 +/- 3.6% predicted). TwQ fell significantly postexercise; 79.2 +/- 5.4% of baseline value at 10 min postexercise (p < 0.005), 75.7 +/- 4.8% at 30 min postexercise (p < 0.001), and 84.0 +/- 5.0% at 60 min postexercise (p < 0.005). Acceptable M-waves from the quadriceps muscle (not obscured by stimulus artifact) were obtained in six subjects. M-wave amplitude was unchanged from baseline at all times postexercise indicating that the fall in TwQ was due to contractile fatigue and not to transmission failure. In conclusion, contractile fatigue of the quadriceps muscle occurs after high intensity cycle exercise to the limits of tolerance in patients with COPD. PMID- 10673185 TI - Alveolar fibrin formation caused by enhanced procoagulant and depressed fibrinolytic capacities in severe pneumonia. Comparison with the acute respiratory distress syndrome. AB - Changes in the alveolar hemostatic balance in severe pneumonia were compared with those in the acute respiratory distress syndrome (ARDS). Analysis was performed in bronchoalveolar lavage fluids (BALF) of patients with ARDS triggered by nonpulmonary underlying events in the absence of lung infection (ARDS; n = 25), pneumonia demanding mechanical ventilation (PNEU-vent; n = 114), spontaneously breathing patients with pneumonia (PNEU-spon; n = 40), and ARDS in combination with lung infection (ARDS+PNEU; n = 43); comparison with healthy control subjects (n = 35) was performed. In all groups of patients, BALF total procoagulant activity was increased by nearly two orders of magnitude, being largely attributable to the tissue factor pathway of coagulation. Concomitantly, markedly reduced overall fibrinolytic capacity (fibrin plate assay) was noted in the lavage fluids of all patients. BALF levels of urokinase-type plasminogen activator were significantly reduced throughout, whereas the lavage concentrations of tissue-type plasminogen activator did not differ from those in control subjects. In addition, markedly enhanced levels of plasminogen activator- inhibitor I and alpha(2)-antiplasmin were noted in ARDS, ARDS+PNEU, and PNEU vent, but not in PNEU-spon. In all groups of patients, the changes in the lavage enzymatic activities were paralleled by manifold increased BALF concentrations of fibrinopeptide A and D-dimer, reflecting in vivo coagulation processes. Within the overall number of patients with pneumonia, changes in the alveolar hemostatic balance were more prominent in alveolar and interstitial pneumonia than in bronchopneumonia. Acute inflammatory lung injury, whether triggered by nonpulmonary systemic events or primary lung infection, is thus consistently characterized by both enhanced procoagulant and depressed fibrinolytic activities in the alveolar lining layer, with the appearance of fibrin formation in this compartment. Profile and extent of changes in severe pneumonia demanding respirator therapy are virtually identical to those in ARDS, whereas somewhat less prominent alterations of the alveolar hemostatic balance are noted in spontaneously breathing patients with pneumonia. PMID- 10673186 TI - Effects of decreased respiratory frequency on ventilator-induced lung injury. AB - To determine if decreased respiratory frequency (ventilatory rate) improves indices of lung damage, 17 sets of isolated, perfused rabbit lungs were ventilated with a peak static airway pressure of 30 cm H(2)O. All lungs were randomized to one of three frequency/peak pulmonary artery pressure combinations: F20P35 (n = 6): ventilatory frequency, 20 breaths/min, and peak pulmonary artery pressure, 35 mm Hg; F3P35 (n = 6), ventilatory frequency, 3 breaths/min, and peak pulmonary artery pressure of 35 mm Hg; or F20P20 (n = 5), ventilatory frequency, 20 breaths/min, and peak pulmonary artery pressure, 20 mm Hg. Mean airway pressure and tidal volume were matched between groups. Mean pulmonary artery pressure and vascular flow were matched between groups F20P35 and F3P35. The F20P35 group showed at least a 4.5-fold greater mean weight gain and a 3-fold greater mean incidence of perivascular hemorrhage than did the comparison groups, all p /= 80% predicted, ratio of FEV(1) to vital capacity [FEV(1)/VC] >/= 80%, methacholine PC(20) >/= 16 mg/ml). Forty-six were atopic. Sputum induction produced an adequate sample in 96 subjects [53 males, mean age (range) 36 (18 to 60) yr]. The expectorate was processed within 2 h; sputum was selected, treated with dithiothreitol, filtered, and examined in a hemocytometer for total cell count and viability and on Wright-stained cytospins for a differential cell count. The mean, median (90th percentile) total cell count was 4.1, 2.4 (9.7) x 10(6) cells/g and cell viability was 69.6, 72.0 (89.7)%. The proportions of eosinophils were 0.4, 0.0 (1.1)%, neutrophils 37.5, 36.7 (64.0)%, macrophages 58.8, 60.8 (86.1)%, lymphocytes 1.0, 0.5 (2.6)%, metachromatic cells 0.0, 0.0 (0.04)%, and bronchial epithelial cells 1.6, 0.3 (4.4)%, respectively. Female gender and atopy were associated with a significant elevation of eosinophils; mean difference between male/female was 0.3% (p = 0.043) and between atopic/nonatopic 0.4% (p = 0.024). This study has identified reference values for total and differential cell counts in induced sputum of healthy adults. PMID- 10673189 TI - No effect of inhaled budesonide on the response to inhaled ozone in normal subjects. AB - Inhalation of ozone in normal subjects causes a neutrophilic inflammatory response in the airways. Pretreatment with inhaled corticosteroids reduces the inflammatory response to inhaled ozone in dogs. We undertook a double-blind, randomized, placebo-controlled, crossover study to investigate the effects of 2 wk of treatment with inhaled budesonide 800 microg twice daily or placebo prior to ozone exposure in humans. Fifteen (six male; mean age, 31.1 +/- 2.1 yr) healthy nonsmokers were exposed to 400 parts per billion (ppb) ozone for 2 h with intermittent exercise. Spirometry, exhaled carbon monoxide (CO) and nitric oxide (NO) levels, measurement of methacholine reactivity, and collection of exhaled air condensate and induced sputum samples were performed at baseline, preexposure, and at intervals up to 24 h postexposure. Ozone exposure led to significant decreases in spirometry and increased methacholine reactivity and sputum neutrophils and myeloperoxidase (MPO). There were no changes in exhaled NO and CO levels, or exhaled breath nitrite after ozone exposure. There were no differences in any of the parameters after treatment with budesonide compared with placebo, and no differences in the response to ozone between treatment groups. We conclude that a high dose of inhaled corticosteroid does not protect against the effects of ozone exposure in normal subjects. PMID- 10673190 TI - Clinical correlates and prognostic significance of six-minute walk test in patients with primary pulmonary hypertension. Comparison with cardiopulmonary exercise testing. AB - The six-minute walk test is a submaximal exercise test that can be performed even by a patient with heart failure not tolerating maximal exercise testing. To elucidate the clinical significance and prognostic value of the six-minute walk test in patients with primary pulmonary hypertension (PPH), we sought (1) to assess the relation between distance walked during the six-minute walk test and exercise capacity determined by maximal cardiopulmonary exercise testing, and (2) to investigate the prognostic value of the six-minute walk test in comparison with other noninvasive parameters. The six-minute walk test was performed in 43 patients with PPH, together with echocardiography, right heart catheterization, and measurement of plasma epinephrine and norepinephrine. Symptom-limited cardiopulmonary exercise testing was performed in a subsample of patients (n = 27). Distance walked in 6 min was significantly shorter in patients with PPH than in age- and sex-matched healthy subjects (297 +/- 188 versus 655 +/- 91 m, p < 0. 001). The distance significantly decreased in proportion to the severity of New York Heart Association functional class. The distance walked correlated modestly with baseline cardiac output (r = 0.48, p < 0.05) and total pulmonary resistance (r = -0.49, p < 0. 05), but not significantly with mean pulmonary arterial pressure. In contrast, the distance walked correlated strongly with peak V O(2) (r = 0.70, p < 0.001), oxygen pulse (r = 0.57, p < 0.01), and V E-VCO(2) slope (r = -0.66, p < 0.001) determined by cardiopulmonary exercise testing. During a mean follow-up period of 21 +/- 16 mo, 12 patients died of cardiopulmonary causes. Among noninvasive parameters including clinical, echocardiographic, and neurohumoral parameters, only the distance walked in 6 min was independently related to mortality in PPH by multivariate analysis. Patients walking < 332 m had a significantly lower survival rate than those walking farther, assessed by Kaplan-Meier survival curves (log-rank test, p < 0.01). These results suggest that the six-minute walk test, a submaximal exercise test, reflects exercise capacity determined by maximal cardiopulmonary exercise testing in patients with PPH, and it is the distance walked in 6 min that has a strong, independent association with mortality. PMID- 10673191 TI - Chlamydia pneumoniae serology, lung function decline, and treatment for respiratory disease. AB - Associations have been reported between Chlamydia pneumoniae seropositivity and both acute and chronic obstructive airway diseases. Plasma specimens collected between 1979 and 1983 from 1, 773 men 45 to 59 yr of age in Caerphilly, South Wales, were tested for IgG and IgA antibodies to C. pneumoniae (TW183) by microimmunofluorescence. Subsequent mortality and medication for obstructive airway disease were ascertained at 5-yr follow-up examinations. Spirometry was performed at the first and second examinations and analyzed both cross sectionally and longitudinally; 642 men (36%) had IgG antibodies at a titer of 1:16 or above, of whom 362 also had detectable IgA antibodies. No statistically significant associations were found between either IgG titer or IgA titer and any of the outcome measures: inhaler therapy at entry; commencement of inhalers during follow-up; death from respiratory causes; baseline FEV(1), FVC, and FEV(1)/FVC ratio; and decline in FEV(1) (p > 0.1 throughout). Men with high IgG titers (>/= 1:64) had a slower rate of decline of FEV(1) than did seronegative subjects (adjusted mean difference in 5-yr change in FEV(1): +22 ml, 95% confidence interval: -31 ml to +76 ml). Men with high IgA titers (>/= 1:16) had a slightly faster rate of decline (-12 ml, - 96 ml to +71 ml). This first prospective assessment suggests that chronic C. pneumoniae infection is not a major risk factor for progressive airflow obstruction. PMID- 10673192 TI - Symptoms, quality of life, and health service contact among young adults with mild asthma. AB - This report assesses Quality of Life (QoL) and its relationship to current symptoms and prospective medical contact among 396 adult patients with asthma. Patients were 16 to 52 yr of age and in the care of family physicians in the northeast of Scotland. All patients had been prescribed asthma medication within the previous 3 mo. Mean %pred FEV(1) was 87.4, mean %pred PEF was 85.1; 41% reported respiratory symptoms every week in the month before interview. Patients completed the SF-36, SF-12, and St. George's Respiratory Questionnaire (SGRQ) scales. Although mean scores on the SF-36 and SF-12 were close to population norms for patients without chronic illness, the presence of any respiratory symptoms in the month before interview was related to significantly lower QoL scores on the SF-36 scales of Physical Functioning, Energy, Mental Health, Pain, and Health Perception: the SF-12 Physical Functioning scale, and the SGRQ Symptoms, Impact and Activities scales. Physician contact for asthma in the 12 mo after interview was significantly related to SF-36, SF-12, and SGRQ scores at time of interview; however, when adjusted for symptoms at time of interview, only the SGRQ scales remained significant predictors of prospective physician contact. We conclude that respiratory symptoms have significant impact on QoL among patients with mild asthma, measured by generic and respiratory QoL scales, but that a specific respiratory scale is better able to discriminate patients who will seek physician care for asthma. PMID- 10673193 TI - Asthma mortality in U.S. Hispanics of Mexican, Puerto Rican, and Cuban heritage, 1990-1995. AB - We used national vital statistics data for 1990 through 1995 to examine both national and regional age-adjusted asthma mortality rates for U.S. Hispanics of Mexican, Cuban, and Puerto Rican heritage, as well as for non-Hispanic whites and non-Hispanic blacks. Nationally, Puerto Ricans had an age-adjusted annual asthma mortality rate of 40.9 per million, followed by Cuban-Americans (15. 8 per million) and Mexican-Americans (9.2 per million). In comparison, non-Hispanic whites had an age-adjusted annual asthma mortality rate of 14.7 per million and non-Hispanic blacks had a rate of 38.1 per million. Age-adjusted asthma mortality for Puerto Ricans was highest in the Northeast (47.8 per million); this region accounted for 81% of all asthma deaths among Puerto Ricans in the United States. In the U.S., Puerto Ricans had the highest asthma mortality rates among Hispanics, followed by Cuban-Americans and Mexican-Americans. In addition, among Hispanic national groups, mortality rates were consistently higher in the Northeast than the Midwest, South, or West regions. These results further support that Hispanics do not represent a uniform, discrete group in terms of health outcomes, and that further public health research and interventions should take Hispanic national origin into account. PMID- 10673194 TI - Response of the canine inspiratory intercostal muscles to chest wall vibration. AB - High-frequency mechanical vibration of the rib cage reduces dyspnea, but the effect of this procedure on the respiratory muscles is largely unknown. In the present studies, we have initially assessed the electrical and mechanical response to vibration (40 Hz) of the canine parasternal and external intercostal muscles (third interspace) during hyperventilation-induced apnea. When the vibrator was applied to the segment investigated, prominent external intercostal activity was recorded in the seven animals studied, whereas low-amplitude parasternal intercostal activity was recorded in only four animals. Similarly, when the vibrator was applied to more rostral and more caudal interspaces, activity was recorded commonly from the external intercostal but only occasionally from the parasternal. The two muscles, however, showed similar changes in length. We next examined the response to vibration of the muscles in seven spontaneously breathing animals. Vibrating the rib cage during inspiration (in-phase) had no effect on parasternal intercostal inspiratory activity but induced a marked increase in neural drive to the external intercostals. For the animal group, peak external intercostal activity during the control, nonvibrated breaths averaged (mean +/- SE) 43.1 +/- 3.7% of the activity recorded during the vibrated breaths (p < 0.001). External intercostal activity during vibration also occurred earlier at the onset of inspiration and commonly carried on after the cessation of parasternal intercostal activity. Yet tidal volume was unchanged. Vibrating the rib cage during expiration (out-of-phase) did not elicit any parasternal or external intercostal activity in six animals. These observations thus indicate that the external intercostals, with their larger spindle density, are much more sensitive to chest wall vibration than the parasternal intercostals. They also suggest that the impact of this procedure on the mechanical behavior of the respiratory system is relatively small. PMID- 10673195 TI - Impaired hepatic extraction and increased splanchnic production contribute to lactic acidosis in canine sepsis. AB - In septic shock, the extent to which lactic acidosis (LA) is a consequence of splanchnic lactate overproduction (SLP) or impaired hepatic lactate extraction (HLE) is not clear. We examined SLP and HLE in E. coli sepsis in dogs. We further determined the effects of vasopressor treatments, which included phenylephrine, dopamine, norepinephrine, and a combination of dobutamine and norepinephrine treatment, on SLP and HLE in respective groups. The animals were studied while anesthetized and ventilated. During sepsis, SLP increased as compared with presepsis (-0.017 versus 0.07 mmol/min, p < 0.05), but this increase could not be explained by reduced splanchnic oxygen delivery (SOD). During sepsis, HLE increased as compared with baseline (0.8 versus 8%, p < 0.05), but was significantly lower than that found during lactic acid loading in nonseptic dogs. None of the vasopressor treatments had a detrimental effect on SLP. These results indicate that LA in sepsis occurs secondary to an increase in splanchnic lactate production that is not related to reduced splanchnic oxygen delivery, as well as to a decrease in hepatic lactate extraction. Effects of different vasoactive agents did not alter either splanchnic lactate production or hepatic lactate extraction in this sepsis model. PMID- 10673196 TI - Combined salmeterol 50 microg and fluticasone propionate 250 microg in the diskus device for the treatment of asthma. AB - Three hundred forty-nine patients with asthma previously treated with medium doses of inhaled corticosteroids during a 2-wk, single-blind, run-in period were randomized to treatment with salmeterol 50 microg combined with fluticasone propionate (FP) 250 microg, salmeterol 50 microg, FP 250 microg, or placebo, each given twice daily through a Diskus device for 12 wk. Mean change in FEV(1) at endpoint was significantly (p 70 mm Hg, p < 0.05), and reduced Q S/Q T (15, 19, and 17%, respectively, p < 0.05) at 6 and 12 h, but not in the Control group. The SNO group had the highest values of Cdyn at 12 h, alveolar aeration and disaturated phosphatidylcholine-to-total protein ratio in bronchoalveolar lavage fluid, and the lowest wet-to-dry lung weight ratio and lung injury score (p < 0.05). The results indicate that early alleviation of ALI by surfactant, iNO, and PSV is due to synergistic effects, and only PSV in this model had limited effects. PMID- 10673204 TI - Natural autoantibody to MUC1 is a prognostic indicator for non-small cell lung cancer. AB - A great deal of attention has been focused on the antitumor effects of anti-MUC1 humoral and cellular responses. We examined whether anti-MUC1 antibody is present in patients with lung cancer, and evaluated its prognostic value. Serum was obtained from 30 patients with nonresectable, non-small cell lung cancer (NSCLC) and 60 healthy volunteers. The presence of anti-MUC1 antibody was determined by enzyme-linked immunosorbent assay. The patients were observed for a median follow up time of 54.0 mo. Overall survival was estimated by the Kaplan-Meier method. Multivariate analyses were performed using the Cox proportional hazards regression model. Anti-KL-6/MUC1 antibody levels of the patients were significantly lower than those of normal individuals (p < 0.001). One-year survival rate of patients with high concentrations of anti-KL-6/MUC1 antibody was significantly higher than that of patients with low levels of anti-KL-6/MUC1 antibody (90.9% versus 21.1%, p < 0.001). Anti-KL-6/MUC1 antibody status was most strongly correlated with mortality, followed by lymph node status and albumin levels, whereas sex, serum lactate dehydrogenase (LDH), and carcinoembryonic antigen (CEA) levels, and metastasis status did not correlate with mortality. These preliminary results indicate that the degree of decrease in antibody level may be associated with a patient's prognosis. PMID- 10673205 TI - Talc induces apoptosis in human malignant mesothelioma cells in vitro. AB - Pleurodesis with talc is an accepted method for the treatment of symptomatic pleural effusions secondary to mesotheliomas. Patients with mesothelioma who have talc-induced pleurodesis have a lower morbidity than do those who do not have pleurodesis. The mechanisms whereby talc mediated these effects were considered to be secondary to a decrease or absence of a pleural effusion. The possibility that talc may directly affect malignant cells was not considered. The present study was designed to evaluate if talc directly effects cell death of malignant mesothelioma cells (MMC) or normal pleural mesothelial cells (PMC). Three confluent MMC and PMC were exposed to talc for 24, 48, and 72 h. In parallel experiments, glass beads similar in size to talc were included as control. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and DNA electrophoresis. Our results demonstrated that talc at a therapeutically achievable concentration (6 microg/cm(2)) induces significant apoptosis in MMC. Talc-induced maximum apoptosis in MMC (39.50 +/- 2.55%, 31.87 +/- 4.69%, and 15.10 +/- 3.93% in CRL 2081, CRL-5820, and CRL-5915, respectively) at 48 h, which was significantly (p < 0.05) greater than that in control cells. Electrophoresis of DNA isolated from talc-exposed MMC demonstrated the typical ladder pattern of internucleosomal DNA cleavage. Talc did not induce apoptosis in PMC, and glass beads did not cause significant apoptosis in either MMC or PMC. The present study has demonstrated that talc induces apoptosis in MMC without affecting normal mesothelial cells of the pleura. PMID- 10673206 TI - The utility of transbronchial needle aspiration in the staging of bronchogenic carcinoma. AB - We conducted a prospective multi-institutional clinical study involving community hospitals and academic medical centers to more carefully define the value of computerized tomography (CT) of the chest with transbronchial needle aspiration (TBNA) in the staging of bronchogenic carcinoma (CA), and to assess the predictors of a positive aspirate. Of 360 individuals determined to have bronchogenic carcinoma, 50 of 81 (62%) with small cell carcinoma (SCC) and 135 of 279 (48%) with non-small cell carcinoma (NSCC) had positive aspirates (p = 0.034). TBNA precluded additional thoracic surgery in a total of 104 of 360 (29%) patients and was exclusively diagnostic of carcinoma in 65 of 360 (18%) cases. Right-sided tumors were more likely to have a positive mediastinal TBNA (p = 0.002 to 0. 01) as were histologic (67 of 118 [57%]) rather than cytology aspirates (228 of 532 [41%]) (p < 0.001). Sensitivity was > 57% in lymph nodes (LN) >/= 10 mm, and among LN of equivalent size, right paratracheal and subcarinal sites were most likely to establish malignancy. Preoperative CT is a valuable adjunct in the staging of CA by TBNA. Increasing LN size, right-sided tumors, right paratracheal and subcarinal locations, use of a histology needle, and the presence of SCC are the best predictors of a positive aspirate. PMID- 10673207 TI - Deficiency of lamellar bodies in alveolar type II cells associated with fatal respiratory disease in a full-term infant. AB - We report a case of a full-term female infant who presented with severe respiratory distress shortly after birth and died at 23 d of age with unremitting respiratory failure. Infectious and other known causes of respiratory disease in this clinical setting were excluded. Examination of a lung biopsy showed abnormal lung parenchyma with features reminiscent of desquamative interstitial pneumonitis. Ultrastructural studies revealed that alveolar type II cells lacked cytoplasmic lamellar bodies, while other organelles appeared normal. Histochemical and immunohistochemical investigations indicated normal alveolar type II cell marker expression including surfactant proteins (SP-A, SP-B, pro-SP B, and pro-SP-C). Mutations in the coding sequences of the SP-B gene were excluded as a cause of disease. This case appears to be a novel congenital defect affecting the pulmonary surfactant system. The cellular abnormality may involve the assembly of cytoplasmic lamellar bodies in alveolar type II cells-the principal storage site of pulmonary surfactant. PMID- 10673208 TI - Manganese superoxide dismutase and catalase are coordinately expressed in the alveolar region in chronic interstitial pneumonias and granulomatous diseases of the lung. AB - Free radicals have been suggested to play an important role in the pathogenesis of interstitial lung diseases, the most important of which are chronic interstitial pneumonias such as usual interstitial pneumonia (UIP) and desquamative interstitial pneumonia (DIP) and granulomatous lung diseases such as sarcoidosis. Because manganese superoxide dismutase (MnSOD) and catalase are two important intracellular antioxidant enzymes that probably play a central role in lung defense, the localization and intensity of these two enzymes were assessed by immunohistochemistry in biopsies of UIP (n = 9), DIP (n = 11), pulmonary sarcoidosis (n = 14), and extrinsic allergic alveolitis (n = 6). The mRNA of these enzymes in selected samples of bronchoalveolar lavage was assessed by Northern blotting. Catalase, but not MnSOD, was constitutively expressed, especially in type II pneumocytes of the healthy lung of nonsmoking individuals. In contrast, manganese SOD immunoreactivity was markedly upregulated in all of the interstitial lung diseases investigated, whereas no increased expression of catalase could be detected in any case. Both enzymes were expressed, especially in type II pneumocytes and alveolar macrophages of DIP and UIP, in the well preserved areas of the lung, in the acute fibromyxoid lesions of UIP, and in the granulomas of sarcoidosis and extrinsic allergic alveolitis. The simultaneous expression of MnSOD and catalase in the alveolar region suggests their protective role against the progression of lung disease. PMID- 10673209 TI - Prolonged allergen exposure induces structural airway changes in sensitized rats. AB - The pathogenesis and functional consequences of airway remodeling in asthma remain to be fully established. In the present study we evaluated the effect of prolonged allergen exposure on airway function and structure in rats. Sensitized Brown Norway rats were repeatedly exposed for periods of 2, 4, or 12 wk to aerosolized ovalbumin (OA) or phosphate-buffered saline (PBS). OA exposure induced a persistent increase in OA-specific serum IgE and in the number of peribronchial eosinophils. After 2 wk of OA exposure, airway histology revealed goblet-cell hyperplasia, an increase in bromodeoxyuridine-positive cells in airway epithelium, increased fibronectin deposition, and a thickening of the airway inner wall area. This coincided with airway hyperresponsiveness (AHR) to aerosolized carbachol. After OA exposure for 12 wk, increased fibronectin (p < 0.05 versus PBS) and collagen deposition (p < 0.05 versus PBS) were observed in the submucosa. After 12 wk of exposure, neither total nor inner wall area or airway responsiveness to carbachol were any longer significantly different from those of PBS-exposed animals. In conclusion, prolonged OA exposure in rats induces structural airway changes that bear similarities to airway remodeling in asthma. The study data further indicate that depending on the extent and distribution of remodeling, changes in the extracellular matrix can enhance or protect against AHR. PMID- 10673210 TI - Prostaglandin H synthase 2 expression in airway cells from patients with asthma and chronic obstructive pulmonary disease. AB - Products of the prostaglandin H synthase (PGHS) metabolic pathway are thought to play a role in the pathogenesis of asthma. We determined the level of expression of the constitutive (PGHS-1) and inducible (PGHS-2) isoforms of the enzyme in induced sputum and bronchial biopsies of patients with asthma, patients with chronic obstructive pulmonary disease (COPD), and unaffected control subjects by immunocyto- and immunohistochemistry. Immunoreactivity for PGHS-2 was significantly greater in the induced sputum of patients with asthma and patients with COPD compared with unaffected control subjects. The level of PGHS-2 was greater in asthma than in COPD. Immunoreactivity for PGHS-1 increased in cells in the induced sputum of patients with asthma and patients with COPD compared with that of unaffected control subjects. Immunostained cells included macrophages, eosinophils, and neutrophils. Greater PGHS-2 immunoreactivity was seen in the submucosal inflammatory infiltrate and in the airway epithelium of patients with asthma compared with unaffected control subjects. In summary, we demonstrate an induction of PGHS-2 in asthma, suggesting increased formation of prostanoids, which may contribute to the inflammatory process. PMID- 10673211 TI - Mycobacterium abscessus infection in cystic fibrosis. Colonization or infection? AB - We present a case of a patient with cystic fibrosis who was thought to be colonized with Mycobacterium abscessus for 13 yr prior to developing clinically apparent mycobacterial infection. However, histologic evidence indicated that invasive mycobacterial disease was present from the onset. While accepting that chronic endobronchial colonization with atypical mycobacteria may occur in patients with cystic fibrosis, the repeated isolation of mycobacteria from the sputum of these patients should alert the clinician to the possibility of indolent disease. Early consideration of treatment for this infection should occur in any patient with cystic fibrosis in whom there is an unexplained deterioration in lung function. The recent introduction of high dose ibuprofen raises concerns about its possible contribution to the progression of the infection. PMID- 10673212 TI - American Thoracic Society. Idiopathic pulmonary fibrosis: diagnosis and treatment. International consensus statement. American Thoracic Society (ATS), and the European Respiratory Society (ERS). PMID- 10673213 TI - American Thoracic Society. What constitutes an adverse health effect of air pollution? Official statement of the American Thoracic Society. PMID- 10673214 TI - Relationship between resting hypercapnia and physiologic parameters before and after lung volume reduction surgery in severe chronic obstructive pulmonary disease. PMID- 10673215 TI - Nocturnal oxygenation and prognosis in Duchenne muscular dystrophy. PMID- 10673216 TI - Leukotrienes as targets for treatment of asthma and other diseases. Current basic and clinical research PMID- 10673217 TI - The discovery of the leukotrienes. PMID- 10673218 TI - From bench to bedside. The hurdles of discovering a new leukotriene receptor antagonist. PMID- 10673219 TI - The molecular biology and regulation of 5-lipoxygenase. PMID- 10673220 TI - Leukotriene C4 synthase. A pivotal enzyme in the biosynthesis of the cysteinyl leukotrienes. PMID- 10673221 TI - Membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG). A widespread protein superfamily. AB - The members of the MAPEG superfamily have been aligned and found to be distantly related, with a common pattern of hydropathy. Figure 2A shows the multiple sequence alignments of the human members and Figure 2B the corresponding superimposed hydropathy profiles. The alignment in Figure 2A demonstrates a total of six strictly conserved residues. The Arg-51 in LTC4 synthase has been suggested to function as proton donor for the opening of the LTA4 epoxide. This arginine is found in all but the FLAP sequences in accordance with the observation that FLAP has no known enzyme activity. Also the Tyr-93 in LTC4 synthase has been suggested to function as a base for the formation of the thiolate anion of glutathione. This tyrosine is not conserved in MGST1 or MGST1 L1. Table 1 summarizes some other properties of the individual human proteins. They are all of the same size, ranging from 147 to 161 amino acids. Only FLAP differs in that its isoelectric point is more neutral than that of the other, more basic proteins. The genes encoding these proteins all reside on different chromosomes (when known) (Table 1). In addition to the human proteins, MAPEG members have been identified in plants, fungi, and bacteria. It is clearly a challenge to elucidate their role in these different phyla in relation to their defined physiological functions in humans. PMID- 10673222 TI - Structure, function, and regulation of leukotriene A4 hydrolase. PMID- 10673223 TI - Phospholipase A2 in eicosanoid generation. PMID- 10673224 TI - Intracellular compartmentalization of leukotriene biosynthesis. PMID- 10673225 TI - Pharmacological characterization of leukotriene receptors. PMID- 10673226 TI - Binding to cysteinyl-leukotriene receptors. PMID- 10673227 TI - Leukotriene B4 receptor. Cloning and intracellular signaling. PMID- 10673228 TI - Cloning of a novel chemoattractant receptor activated by leukotriene B4 and used by human immunodeficiency virus type 1 to infect CD4-positive immune cells. A therapeutic connection to asthma? PMID- 10673230 TI - Leukotriene bronchoconstriction induced by allergen and exercise. PMID- 10673229 TI - The role of leukotrienes in the regulation of tone and responsiveness in isolated human airways. AB - Cysteinyl-leukotrienes and histamine are the major determinants of inherent tone in isolated human bronchi, which is mainly the result of a balance of continual production and release of contractile mediator, in particular cysteinyl leukotrienes and to a lesser extent histamine, and on the other side bronchodilating prostanoids. Cysteinyl-leukotrienes are also powerful constrictors of isolated human airways through direct interaction with Cys-LT1 receptors on airway smooth muscle, and with a potency 1,000-fold higher than histamine. On stimulation inflammatory cells such as eosinophils and mst cells produce and release significantly increased amounts of leukotrienes leading to smooth muscle contration in vitro. In isolated human airways, leukotrienes are the most important mediators of allergen and adenosine-induced contractile responses. The induction of allergen responses in passively sensitized airways is not only related to an increased release of leukotrienes and histamines, but also to an enhanced responsiveness of the airway smooth muscle, particularly to LTC4. Studies in isolated human airways in vitro have demonstrated that understanding the regulation of human airway tone and airway reactivity are closely linked to the understanding of baseline and stimulated production of and smooth muscle responsiveness to leukotrienes in vitro and in vivo. PMID- 10673231 TI - Effects of antileukotrienes in the treatment of asthma. PMID- 10673232 TI - Genetic variations in the 5-lipoxygenase core promoter. Description and functional implications. PMID- 10673233 TI - The metabolism of leukotriene B4 in Lewis lung carcinoma porcine kidney cells. PMID- 10673234 TI - Adenosine, a potent natural suppressor of arachidonic acid release and leukotriene biosynthesis in human neutrophils. PMID- 10673235 TI - Formation of endogenous "antiinflammatory" lipid mediators by transcellular biosynthesis. Lipoxins and aspirin-triggered lipoxins inhibit neutrophil recruitment and vascular permeability. PMID- 10673236 TI - Measurement of leukotrienes in humans. PMID- 10673237 TI - Functional studies of leukotriene receptors in vascular tissues. AB - The paradoxical effects of cysteinyl-leukotrienes, namely contraction and relaxation, are now well documented in a number of vascular preparations from various species. The vascular smooth muscle contractions are associated with activation of a single receptor subtype and in some vascular smooth muscles with activation of two receptor subtypes. However, the receptors implicated in the contraction of vessels such as pig pulmonary arteries and veins, dog inferior vena cava, and dog splenic and mesenteric veins remain to be established. There are sufficient data concerning some vascular tissues to suggest that relaxations induced by cysteinyl-leukotrienes are via the stimulation of specific receptors present on the endothelium. The endothelium in human pulmonary arteries has one receptor (CysLT2) and activation induced the release of NO. However, in isolated human pulmonary veins two receptors are present, CysLT1 and CysLT2 (Figure 1). Activation of the former induced the release of a contractile factor whereas activation of the CysLT2 receptor released NO. In guinea pig pulmonary artery and guinea pig thoracic aorta, one receptor has been demonstrated since the relaxations are blocked by ICI-198615. These data suggest the presence of a CysLT1 receptor. Activation of this receptor leads to the release of a relaxant factor, namely, nitric oxide. In contrast, in human pulmonary arteries and veins activation of a receptor that is resistant to ICI-198615 is associated with NO release. These results suggest that there may be species differences even when analogous vascular preparations are examined. While the cysteinyl-leukotrienes are known to relax vascular smooth muscle in a variety of preparations from different species, there are presently two pathways known to be involved in this response. One involves the metabolites of arachidonic acid via the cyclooxygenase enzymatic pathway and the other implicates products of the L-arginine enzymatic pathway. Although both pathways may be present and active in the endothelium of the vascular preparations only one of these enzymatic pathways may be dominant and responsible for the relaxations observed. Ortiz and coworkers have demonstrated that in pulmonary veins the dominant pathway for cysteinyl leukotriene relaxations is the NO pathway. There are some reports from animal studies that support a dominant role for NO in pulmonary veins. In contrast, Allen and co-workers demonstrated that the LTC4-induced relaxations in isolated human saphenous veins were not modified by treatment of tissues with an NO inhibitor but were significantly enhanced after treatment with indomethacin. These authors suggested that a contracting factor derived from the arachidonic acid pathway was released in preparations challenged with LTC4. In addition, these investigators demonstrated that the NO inhibitor had no effect on the LTC4 relaxations. Together, these results suggest that cysteinyl-leukotriene effects in human pulmonary veins are dominated by the NO pathway whereas in human systemic veins these mediator effects are modified by metabolites of the cyclooxygenase pathway. Unfortunately, most studies involving the actions of cysteinyl-leukotrienes on vessels have been performed in the presence of indomethacin, making interpretation of the relative contribution of the cyclooxygenase and NO pathways difficult. In any event, the cysteinyl leukotrienes may have a prominent role in the activation of these pathways and the receptors involved have not been clearly established. PMID- 10673238 TI - Leukotrienes in cardiovascular diseases. PMID- 10673239 TI - Interactions between leukotrienes and other inflammatory mediators/modulators in the microvasculature. PMID- 10673240 TI - 5-Lipoxygenase and leukotrienes. Transgenic mouse and nuclear targeting studies. PMID- 10673241 TI - Role of leukotrienes in bronchial hyperresponsiveness and cellular responses in airways. PMID- 10673242 TI - Leukotrienes in rhinitis. PMID- 10673243 TI - Treatment of aspirin-intolerant asthma with antileukotrienes. PMID- 10673244 TI - Genetic mechanisms in aspirin-induced asthma. PMID- 10673246 TI - Circulation online only : february 15, 2000 PMID- 10673245 TI - Antileukotriene therapy. Future directions. PMID- 10673247 TI - Serial in vivo MRI documents arterial remodeling in experimental atherosclerosis. AB - BACKGROUND: Arterial remodeling in response to atherosclerosis may be outward (positive) or inward (negative) and is an important mechanism in the clinical manifestations of atherosclerosis and restenosis after percutaneous coronary interventions. Postmortem and intravascular ultrasound studies of arterial remodeling do not allow serial and noninvasive data to be obtained. In a rabbit model of atherosclerosis, we sought to validate MRI as a new tool for documentation of arterial remodeling. METHODS AND RESULTS: Watanabe heritable hyperlipidemic rabbits underwent serial MRI at baseline and 6 months after aortic balloon denudation. The lumen area had a small but significant (P=0.006) increase, from 4.36+/-0.16 to 4. 89+/-0.12 mm(2). There was a large, significant (P<0.0001) increase in the outer wall area, from 7.96+/-0.19 to 10.46+/-0.19 mm(2). The vessel wall area (a marker of atherosclerotic burden) increased significantly (P<0.0001), from 3.61+/-0.07 to 5.57+/-0.09 mm(2). Thus, the increase in atherosclerotic burden over time was completely accounted for by positive arterial remodeling. The subgroup used for histopathological validation confirmed a significant (P<0.0001) agreement between histopathology and MRI for assessment of all 3 parameters. CONCLUSIONS: MRI can provide serial and noninvasive data about the arterial wall, allowing assessment of arterial remodeling in this rabbit model. Thus, MRI appears to be a useful tool for the investigation of arterial remodeling both in native atherosclerosis and after percutaneous coronary intervention. PMID- 10673248 TI - A randomized comparison of clopidogrel and aspirin versus ticlopidine and aspirin after the placement of coronary-artery stents. AB - BACKGROUND: The introduction of an effective antiplatelet therapy with aspirin and ticlopidine after the placement of coronary-artery stents has decreased the risk of thrombotic stent occlusions (TSO) and hemorrhagic complications. However, the use of ticlopidine is limited by hematological and gastrointestinal adverse effects. The safety and efficacy of clopidogrel after stenting remains to be established. METHODS AND RESULTS: After successful coronary stenting during elective or emergency percutaneous transluminal coronary angioplasty, 700 patients with 899 lesions were randomly assigned to receive a 4-week course of either 500 mg ticlopidine (n=345) or 75 mg clopidogrel (n=355), in addition to 100 mg aspirin. All the following clinical events reflecting TSO were included in the prespecified primary cardiac endpoint: cardiac death, urgent target vessel revascularization, angiographically documented TSO, or nonfatal myocardial infarction within 30 days. The primary noncardiac endpoint was defined as noncardiac death, stroke, severe peripheral vascular or hemorrhagic events, or any adverse event resulting in discontinuation of study medication. Cardiac events occurred in 17 patients [11 (3.1%) with clopidogrel and 6 (1.7%) with ticlopidine (P=0.24)]. The primary noncardiac endpoint was observed in 16 patients (4.5%) assigned to receive clopidogrel versus 33 patients (9.6%) assigned to receive ticlopidine (P=0.01). CONCLUSIONS: After the placement of coronary-artery stents in unselected patients, antiplatelet therapy with aspirin and clopidogrel seems to be comparably safe and effective as aspirin and ticlopidine. Noncardiac events were significantly reduced with clopidogrel. PMID- 10673249 TI - Spironolactone increases nitric oxide bioactivity, improves endothelial vasodilator dysfunction, and suppresses vascular angiotensin I/angiotensin II conversion in patients with chronic heart failure. AB - BACKGROUND: The RALES study showed that spironolactone, added to conventional therapy for chronic heart failure, dramatically reduced mortality. We tested the hypothesis that this benefit was partially due to improvement in endothelial function and/or to amplified suppression of the vascular renin-angiotensin axis. METHODS AND RESULTS: We performed a randomized, placebo-controlled, double-blind crossover study on 10 patients with NYHA class II to III chronic heart failure on standard diuretic/ACE inhibitor therapy, comparing 50 mg/d spironolactone (1 month) versus placebo. Forearm vasculature endothelial function was assessed by bilateral forearm venous occlusion plethysmography using acetylcholine and N monomethyl-L-arginine (L-NMMA), with sodium nitroprusside as a control vasodilator. Also, vascular ACE activity was assessed by use of angiotensin (Ang) I, with Ang II as a control vasoconstrictor. Spironolactone significantly increased the forearm blood flow response to acetylcholine (percentage change in forearm blood flow [mean+/-SEM], 177+/-29% versus 95+/-20%, spironolactone versus placebo; P<0.001), with an associated increase in vasoconstriction due to L-NMMA (-35+/-6% versus -18+/-4%; P<0.05). The Ang I response was also significantly reduced with spironolactone (P<0.05), with Ang II responses unaltered. CONCLUSIONS: Spironolactone improves endothelial dysfunction, increases NO bioactivity, and inhibits vascular Ang I/Ang II conversion in patients with heart failure, providing novel mechanisms for its beneficial effect on cardiovascular mortality. PMID- 10673250 TI - Extent and direction of arterial remodeling in stable versus unstable coronary syndromes : an intravascular ultrasound study. AB - BACKGROUND: The morphological characteristics of coronary plaques in patients with stable versus unstable coronary syndromes have been described in vivo with intravascular ultrasound, but the relationship between arterial remodeling and clinical presentation is not well known. METHODS AND RESULTS: We studied 85 patients with unstable and 46 patients with stable coronary syndromes using intravascular ultrasound before coronary intervention. The lesion site and a proximal reference site were analyzed. The remodeling ratio (RR) was defined as the ratio of the external elastic membrane (EEM) area at the lesion to that at the proximal reference site. Positive remodeling was defined as an RR >1.05 and negative remodeling as an RR <0.95. Plaque area (13.9+/-5.5 versus 11.1+/-4.8 mm(2); P=0.005), EEM area (16.1+/-6.2 versus 13.0+/-4.8 mm(2); P=0. 004), and the RR (1.06+/-0.2 versus 0.94+/-0.2; P=0.008) were significantly greater at target lesions in patients with unstable syndromes than in patients with stable syndromes. Positive remodeling was more frequent in unstable than in stable lesions (51. 8% versus 19.6%), whereas negative remodeling was more frequent in stable lesions (56.5% versus 31.8%) (P=0.001). CONCLUSIONS: Positive remodeling and larger plaque areas were associated with unstable clinical presentation, whereas negative remodeling was more common in patients with stable clinical presentation. This association between the extent of remodeling and clinical presentation may reflect a greater tendency of plaques with positive remodeling to cause unstable coronary syndromes. PMID- 10673251 TI - Atherosclerotic plaque burden and CK-MB enzyme elevation after coronary interventions : intravascular ultrasound study of 2256 patients. AB - BACKGROUND: Elevation of serum creatine kinase MB fraction (CK-MB) after percutaneous coronary interventions has been associated with early and late mortality; however, the pathogenesis of CK-MB elevation is still unknown. We hypothesized that CK-MB elevation was related to atherosclerotic plaque burden as assessed by preintervention intravascular ultrasound (IVUS). METHODS AND RESULTS: We studied 2256 consecutive patients who underwent intervention of 2780 native coronary lesions and had complete high-quality preintervention IVUS imaging in the era before routine use of platelet glycoprotein IIb/IIIa inhibitors. Patients were divided into 3 groups: CK-MB within normal range (1675 patients; 2061 lesions); CK-MB elevation 1 to 5 times upper limit of normal (292 patients; 355 lesions); and CK-MB elevation > or = 5 times upper limit of normal (289 patients; 364 lesions). Qualitative angiographic lesion morphology and quantitative analysis were similar among the 3 groups. On preintervention IVUS, progressively more reference segment and lesion site plaque burden and lesion site calcium occurred in the groups with CK-MB elevation. Positive remodeling was more common in lesions with CK-MB elevation. As levels of CK-MB increased, cross-sectional narrowing (percentage plaque burden) increased, both at the reference site (mean cross-sectional narrowing values were 45.1%, <49.3%, and <52.2% for normal CK-MB, 1 to 5 times upper limit of normal, and > or =5 times upper limit of normal groups, respectively; P=0.03) and at the lesion site (81.9%, <85.4%, and <87.1%, respectively; P=0.04). Multivariate analysis indicated that de novo lesions, atheroablative technique, plaque burden at the lesion and reference segments, and final minimal lumen diameter were independent predictors of CK-MB elevation. CONCLUSIONS: CK-MB elevation correlates with a greater atherosclerotic plaque burden. CK-MB elevation after intervention may be a marker of diffuse atherosclerotic disease or a consequence of catheter-based intervention in more diseased arteries or both. PMID- 10673252 TI - Diastolic blood pressure changes during exercise positively correlate with serum cholesterol and insulin resistance. AB - BACKGROUND: Metabolic factors, including plasma concentrations of cholesterol and insulin resistance, may influence blood pressure through effects on vascular reactivity. Such effects might influence blood pressure during exercise more strongly than at rest. METHODS AND RESULTS: We examined whether there is an association between serum cholesterol or insulin resistance and change in blood pressure during mild exercise. Blood pressure was measured at rest and during fixed low-workload bicycle ergometry (50, 75, and 100 W, each for 3 minutes) in 75 healthy active men (age, 18 to 66 years). Blood pressure at rest was not significantly correlated with serum cholesterol or insulin resistance (estimated from the fasting glucose-insulin product). The change from resting values in diastolic but not systolic blood pressure during exercise was correlated with serum cholesterol (R>0.47, P<0.0001 for each workload) and insulin resistance (R>0.38, P<0.01 for each workload). Serum cholesterol and insulin resistance were the only independent predictors of the change in diastolic blood pressure during exercise in a stepwise regression model incorporating age, body mass index, serum cholesterol, triglycerides, HDL cholesterol, insulin resistance, and heart rate during exercise. In a further study, the change in diastolic blood pressure during exercise was greater in men with uncomplicated type 2 diabetes (13.6 mm Hg [95% CI, 8.5 to 18.8]; n=10) than in nondiabetic control men (2.7 mm Hg [95% CI, 2. 0 to 7.3]; n=10; P=0.002). CONCLUSIONS: Changes in diastolic blood pressure during gentle exercise are strongly associated with serum concentrations of total cholesterol and insulin resistance. This may contribute to development of hypertensive complications in dyslipidemic and/or insulin-resistant patients. PMID- 10673253 TI - Effectiveness and limitations of beta-blocker therapy in congenital long-QT syndrome. AB - BACKGROUND: beta-blockers are routinely prescribed in congenital long-QT syndrome (LQTS), but the effectiveness and limitations of beta-blockers in this disorder have not been evaluated. METHODS AND RESULTS: The study population comprised 869 LQTS patients treated with beta-blockers. Effectiveness of beta-blockers was analyzed during matched periods before and after starting beta-blocker therapy, and by survivorship methods to determine factors associated with cardiac events while on prescribed beta-blockers. After initiation of beta-blockers, there was a significant (P<0.001) reduction in the rate of cardiac events in probands (0.97+/ 1.42 to 0.31+/-0.86 events per year) and in affected family members (0. 26+/-0.84 to 0.15+/-0.69 events per year) during 5-year matched periods. On-therapy survivorship analyses revealed that patients with cardiac symptoms before beta blockers (n=598) had a hazard ratio of 5.8 (95% CI, 3.7 to 9.1) for recurrent cardiac events (syncope, aborted cardiac arrest, or death) during beta-blocker therapy compared with asymptomatic patients; 32% of these symptomatic patients will have another cardiac event within 5 years while on prescribed beta-blockers. Patients with a history of aborted cardiac arrest before starting beta-blockers (n=113) had a hazard ratio of 12.9 (95% CI, 4.7 to 35.5) for aborted cardiac arrest or death while on prescribed beta-blockers compared with asymptomatic patients; 14% of these patients will have another arrest (aborted or fatal) within 5 years on beta-blockers. CONCLUSIONS: beta-blockers are associated with a significant reduction in cardiac events in LQTS patients. However, syncope, aborted cardiac arrest, and LQTS-related death continue to occur while patients are on prescribed beta-blockers, particularly in those who were symptomatic before starting this therapy. PMID- 10673254 TI - Antiarrhythmic efficacy of dipyridamole in treatment of reperfusion arrhythmias : evidence for cAMP-mediated triggered activity as a mechanism responsible for reperfusion arrhythmias. AB - BACKGROUND: Intracellular calcium overload is believed to play an important role in development of reperfusion arrhythmias. Dipyridamole, an inhibitor of cellular uptake of adenosine, may prevent or terminate reperfusion arrhythmias by reducing intracellular calcium overload. METHODS AND RESULTS: First, we tested for a preventive effect of dipyridamole. Sixty-one patients who underwent primary PTCA for treatment of acute anterior wall myocardial infarction were enrolled in this prospective study. Patients were divided into dipyridamole (DP) and nondipyridamole (non-DP) groups. The 2 groups had similar baseline characteristics. In the DP group, dipyridamole 0.5 mg/kg was infused intravenously for 3 minutes immediately before reperfusion during primary PTCA. Arrhythmias after reperfusion were analyzed from continuous ECG recordings. None of the patients in the DP group (n=23) had accelerated idioventricular rhythms (AIVR) or ventricular tachycardia (VT). In contrast, 7 (18.4%) had AIVR and 3 (7.9%) had VT in the non-DP group (n=38; P<0.01). Second, we tested for a termination effect of dipyridamole. Dipyridamole 0.5 mg/kg was infused intravenously while continuous ECG recordings were obtained in 9 patients who had either sustained AIVR (n=7) or sustained VT (n=2) after reperfusion of occluded coronary artery. Arrhythmias were terminated in all patients. CONCLUSIONS: These results indicate that administration of dipyridamole can prevent and terminate reperfusion arrhythmias such as AIVR and VT. cAMP-mediated triggered activity may, at least in part, be responsible for reperfusion-induced AIVR and VT. PMID- 10673255 TI - Dual-loop intra-atrial reentry in humans. AB - BACKGROUND: Dual-loop atrial reentrant tachycardias have not been clinically described. METHODS AND RESULTS: Five patients (3 men, 2 women; mean age, 48+/-16 years) were studied 24+/-15 years after surgical closure of an ostium secundum atrial septal defect for drug-resistant atrial tachycardia. Complete tachycardia mapping was performed in the right atrium with multipolar catheters and a 3 dimensional electroanatomic mapping system (Biosense), followed by linear radiofrequency ablation of the narrowest part of each complete loop. Six tachycardias with a typical flutter morphology, a cycle length of 262+/-40 ms, and a superior f-wave axis (-77+/-11 degrees ) were mapped, 4 with a Biosense map including 106+/-32 points. Five figure-8 tachycardias had a counterclockwise loop around the tricuspid valve sharing a common anterior channel with a clockwise loop around the lateral atriotomy scar. One tachycardia was thought to have 2 counterclockwise loops around the same obstacles. Radiofrequency delivery in the cavotricuspid isthmus in each case transformed the tachycardia without any pause in a different morphology tachycardia with an inferior P-wave axis (50+/-42 degrees ) and nearly the same cycle length (272+/-39 ms) but with the periatriotomy loop alone. This arrhythmia required ablation of a second isthmus: between the lower end of the atriotomy and the inferior vena cava in 4 and the superior tricuspid annulus in 1. After a follow-up of 19+/-6 months, there were no recurrences. CONCLUSIONS: Figure-8 double-loop tachycardias mimicking the ECG pattern of a common atrial flutter occur in some patients after a surgical atriotomy. Ablation of 1 loop produces a sudden transformation to a new reentrant tachycardia formed of the remaining loop that requires ablation at a second isthmus. PMID- 10673256 TI - Analysis by early angiography of right internal thoracic artery grafting via the transverse sinus : predictors of graft failure. AB - BACKGROUND: There has been debate regarding whether technically demanding right internal thoracic artery (RITA) grafting via the transverse sinus can be extensively applied to patients in high-risk groups, such as patients with a small body size, elderly patients, and woman with relatively smaller coronary artery and internal thoracic artery (ITA) diameters. METHODS AND RESULTS: Of the 1456 patients who underwent isolated coronary artery bypass grafting between January 1989 and December 1998 at Kumamoto Central Hospital, 393 patients (mean age, 62.4+/-9.0 years) with the RITA anastomosed to the major branches of the circumflex artery were studied. Left ITA grafting was performed in 384 patients, and in 369, the in situ left ITA was anastomosed to the left anterior descending coronary artery using standard methods. Early postoperative angiography was performed in 381 patients. The RITA was occluded in 4 patients, and string-like artery and significant stenosis were present in 11 and 7 patients, respectively; RITA graft patency was thus 94.1%. Of the preoperative variables and angiographic data, simple and multiple logistic regression analyses identified decreased severity of native stenosis, diffuse sclerosis of native vessels, and residual side branches of the ITA as independent predictors of nonfunctional grafts. The method of ITA grafting did not influence the patency of the graft. CONCLUSIONS: The excellent patency rate demonstrated by this study, the largest angiographic study to date of RITA grafting via the transverse sinus, indicates that this technique can provide reliable revascularization of the left ventricle and that it has the potential to be applied to a wide variety of patients with diseased circumflex arteries. PMID- 10673257 TI - The anatomic basis of connections between the coronary sinus musculature and the left atrium in humans. AB - BACKGROUND: This study determined the histological features of the atrial myocardium connecting the coronary sinus and the left atrium in humans. METHODS AND RESULTS: Ten necropsied hearts were studied by performing serial longitudinal sections parallel to the long axis of the coronary sinus that extended its full length using a large microtome. In all specimens, the venous wall of the coronary sinus was surrounded by a cuff of striated muscle extending 40+/-8 mm from the ostium. Striated myocardial connections of varying number and morphology left this coronary muscle cuff and connected to the left atrium; they ranged from 1 to 2 fascicles to a widely intermingled continuum (thickness, 2.79+/-2 mm; width, 2.91+/-3.5 mm). These connections originated 8.8+/-5.7 mm from the coronary sinus ostium and inserted 18+/-11 mm distally into the left atrium. The insulating compartment in which the connections traversed between the left atrium and the coronary sinus was mostly formed of adipose tissue. The valve of Vieussens was found in 6 hearts at a mean distance of 3.4+/-3.2 mm from the distal extremity of the coronary sinus muscle cuff. CONCLUSIONS: In the human heart, a consistent but morphologically variable left atrial coronary sinus myocardial connection was found. This emphasizes the need for surgical dissection or catheter ablation in or around the coronary sinus to eliminate these connections. PMID- 10673258 TI - Bradycardia and the role of beta-blockade in the amelioration of left ventricular dysfunction. AB - BACKGROUND: It is clear that beta-blockers are effective for treatment of congestive heart failure, but their mechanism of action remains controversial. Hypothesized mechanisms include normalization of beta-receptor function and myocardial protection from the effects of catecholamines, possibly by the institution of bradycardia. We hypothesized that beta-blockade-induced bradycardia was an important mechanism by which these agents were effective for correction of LV dysfunction. METHODS AND RESULTS: In 2 groups of dogs with mitral regurgitation and LV dysfunction, beta-blockers were instituted. In 1 group that received beta-blockers and pacing (group beta+P), a pacemaker prevented the natural bradycardia that beta-blockers cause. In both groups, substantial LV dysfunction developed. Before beta-blockade, the end-systolic stiffness constant decreased from 3. 5+/-0.1 to 2.7+/-0.2 (P<0.01) at 3 months in group beta+P. A similar reduction occurred in the group that eventually received only beta-blockers (group betaB). In group betaB, end-systolic stiffness improved after 3 months of beta-blockade from 2.9+/-0.2 to 3.5+/-0.4 and was not different from baseline. However, in group beta+P, end-systolic stiffness failed to improve (2.7+/-0.2) after 3 months of mitral regurgitation, and was 2.9+/-0.2 at the end of the studies. The contractile function of cardiocytes isolated from the ventricles at the end of the studies confirmed these in vivo estimates of contractility. CONCLUSIONS: We conclude that institution of bradycardia is a major mechanism by which beta-blockers are effective for restoration of contractile function in a model of LV dysfunction. PMID- 10673259 TI - Akt promotes survival of cardiomyocytes in vitro and protects against ischemia reperfusion injury in mouse heart. AB - BACKGROUND: IGF-1 has been shown to protect myocardium against death in animal models of infarct and ischemia-reperfusion injury. In the present study, we investigated the role of the IGF-1-regulated protein kinase Akt in cardiac myocyte survival in vitro and in vivo. METHODS AND RESULTS: IGF-1 promoted survival of cultured cardiomyocytes under conditions of serum deprivation in a dose-dependent manner but had no effect on cardiac fibroblast survival. The cytoprotective effect of IGF-1 on cardiomyocytes was abrogated by the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin. Wortmannin had no effect on cardiomyocyte viability in the absence of IGF-1. IGF-1-mediated cytoprotection correlated with the wortmannin-sensitive induction of Akt protein kinase activity. To examine the functional consequences of Akt activation in cardiomyocyte survival, replication-defective adenoviral constructs expressing wild-type, dominant-negative, and constitutively active Akt genes were constructed. Transduction of dominant-negative Akt blocked IGF-1-induced survival but had no effect on cardiomyocyte survival in the absence of IGF-1. In contrast, transduction of wild-type Akt enhanced cardiomyocyte survival at subsaturating levels of IGF-1, whereas constitutively active Akt protected cardiomyocytes from apoptosis in the absence of IGF-1. After transduction into the mouse heart in vivo, constitutively active Akt protected against myocyte apoptosis in response to ischemia-reperfusion injury. CONCLUSIONS: These data are the first documentation that Akt functions to promote cellular survival in vivo, and they indicate that the activation of this pathway may be useful in promoting myocyte survival in the diseased heart. PMID- 10673260 TI - Microbubble persistence in the microcirculation during ischemia/reperfusion and inflammation is caused by integrin- and complement-mediated adherence to activated leukocytes. AB - BACKGROUND: Albumin microbubbles that are used for contrast echocardiography persist within the myocardial microcirculation after ischemia/reperfusion (I-R). The mechanism responsible for this phenomenon is unknown. METHODS AND RESULTS: Intravital microscopy of the microcirculation of exteriorized cremaster muscle was performed in 12 wild-type mice during intravenous injections of fluorescein labeled microbubbles composed of albumin, anionic lipids, or cationic lipids. Injections were performed at baseline and after 30 to 90 minutes of I-R in 8 mice and 2 hours after intrascrotal tumor necrosis factor-alpha (TNF-alpha) in 4 mice. Microbubble adherence at baseline was uncommon (<2/50 high-power fields). After I R, adherence increased (P<0.05) to 9+/-5 and 5+/-4 per 50 high-power fields for albumin and anionic lipid microbubbles, respectively, due to their attachment to leukocytes adherent to the venular endothelium. TNF-alpha produced even greater microbubble binding, regardless of the microbubble shell composition. The degree of microbubble attachment correlated (r=0.84 to 0.91) with the number of adhered leukocytes. Flow cytometry revealed that microbubbles preferentially attached to activated leukocytes. Albumin microbubble attachment was inhibited by blocking the leukocyte beta(2)-integrin Mac-1, whereas lipid microbubble binding was inhibited when incubations were performed in complement-depleted or heat inactivated serum rather than control serum. CONCLUSIONS: Microvascular attachment of albumin and lipid microbubbles in the setting of I-R and TNF-alpha induced inflammation is due to their beta(2)-integrin- and complement-mediated binding to activated leukocytes adherent to the venular wall. Thus, microbubble persistence on contrast ultrasonography may be useful for the detection and monitoring of leukocyte adhesion in inflammatory diseases. PMID- 10673261 TI - Regulation of endothelial constitutive nitric oxide synthase gene expression in endothelial cells and in vivo : a specific vascular action of insulin. AB - BACKGROUND: The vasodilatory effect of insulin can be acute or increase with time from 1 to 7 hours, suggesting that insulin may enhance the expression of endothelial nitric oxide synthase (eNOS) in endothelial cells. The objective of the present study was to characterize the extent and signaling pathways by which insulin regulates the expression of eNOS in endothelial cells and vascular tissues. METHODS AND RESULTS: Physiological concentrations of insulin (10(-10) to 10(-7) mmol/L) increased the levels of eNOS mRNA, protein, and activity by 2-fold after 2 to 8 hours of incubation in cultured bovine aortic endothelial cells. Insulin enhanced eNOS gene expression in microvessels isolated from Zucker lean rats but not from insulin-resistant Zucker fatty rats. Inhibitors of phosphatidylinositol-3 kinase (PI-3 kinase) decreased the effect of insulin on eNOS gene expression, but a general protein kinase C (PKC) inhibitor, GF109203X or PKCbeta isoform inhibitor, LY333531 enhanced eNOS expression. In contrast, PKC activators inhibited both the activation by insulin of PI-3 kinase and eNOS mRNA levels. Overexpression of PKCbeta isoform in endothelial cells inhibited the stimulation by insulin of eNOS expression and PI-3 kinase activities in parallel. CONCLUSIONS: Insulin can regulate the expression of eNOS gene, mediated by the activation of PI-3 kinase, in endothelial cells and microvessels. Thus, insulin may chronically modulate vascular tone. The activation of PKC in the vascular tissues as in insulin resistance and diabetes may inhibit PI-3 kinase activity and eNOS expression and may lead to endothelial dysfunctions in these pathological states. PMID- 10673263 TI - alpha-adrenergic coronary vasoconstriction and myocardial ischemia in humans. AB - The use of quantitative coronary angiography, combined with Doppler and PET, has recently been directed at the study of alpha-adrenergic coronary vasomotion in humans. Confirming prior animal experiments, there is no evidence of alpha adrenergic coronary constrictor tone at rest. Again confirming prior experiments, responses to alpha-adrenoceptor activation are augmented in the presence of coronary endothelial dysfunction and atherosclerosis, involving both alpha(1)- and alpha(2)-adrenoceptors in epicardial conduit arteries and microvessels. Such augmented alpha-adrenergic coronary constriction is observed during exercise and coronary interventions, and it is powerful enough to induce myocardial ischemia and limit myocardial function. Recent studies indicate a genetic determination of alpha(2)-adrenergic coronary constriction. PMID- 10673262 TI - Antisense inhibition of beta(1)-adrenergic receptor mRNA in a single dose produces a profound and prolonged reduction in high blood pressure in spontaneously hypertensive rats. AB - BACKGROUND: beta-Blockers are the first line of therapy for hypertension. However, they are associated with side effects because of central nervous system (CNS) effects and beta(2)-adrenergic antagonism. To overcome these problems and provide a long-term beta(1)-blockade, antisense oligonucleotides against rat beta(1)-adrenergic receptor (beta(1)-AR) mRNA (beta(1)-AS-ODN) were designed and tested for the ability to inhibit cardiac beta(1)-ARs as well as lower blood pressure in spontaneously hypertensive rats (SHRs). METHODS AND RESULTS: Radioligand binding assay showed that a single intravenous injection of beta(1) AS-ODN delivered in cationic liposomes significantly decreased cardiac beta(1)-AR density by 30% to 50% for 18 days (P<0.01), with no effect on beta(2)-ARs. This was accompanied by marked attenuation of beta(1)-AR-mediated positive inotropic response in isolated perfused hearts in vitro (P<0.02) and in conscious SHRs monitored by telemetry in vivo (P<0.02). Furthermore, the blood pressure of SHRs was reduced for 20 days, with a 38 mm Hg maximum drop. Heart rate was not significantly decreased. Quantitative autoradiography was performed to assess beta(1)-AS-ODN effects on the CNS, which demonstrated no changes in beta(1)-ARs in brain, in contrast to a significant reduction in heart and kidney (P<0.05). For comparison with beta-blockers, the effects of atenolol on cardiovascular hemodynamics were examined, which lowered blood pressure for only 10 hours and elicited appreciable bradycardia in SHRs. CONCLUSIONS: These results indicate that beta(1)-AS-ODN, a novel approach to specific beta(1)-blockade, has advantages over currently used beta-blockers in providing a profound and prolonged reduction in blood pressure without affecting heart rate, beta(2)-ARs, and the CNS. Diminished cardiac contractility resulting from less beta(1)-AR expression contributes to the antihypertensive effect. PMID- 10673264 TI - Images in cardiovascular medicine. Hemolysis after mitral valve repair. PMID- 10673265 TI - Synthetic inhibitors of platelet glycoprotein IIb/IIIa in clinical development. AB - Activation of the platelet glycoprotein (GP IIb/IIIa) receptor on the platelet surface is the final pathway of platelet aggregation, regardless of the initiating stimulus. Inhibitors of GP IIb/IIIa receptors include monoclonal antibodies (abciximab) against this receptor and peptidic and nonpeptidic synthetic specific receptor blockers. Abciximab exchanges between and binds to platelets for as long as 2 weeks, whereas synthetic GP IIb/IIIa inhibitors inhibit ex vivo platelet aggregation for only a few hours after the end of infusion, but some have the advantage of also being orally active. In the secondary prevention of atherothrombosis, large-scale trials were successfully conducted with aspirin, dipyridamole, ticlopidine, and clopidogrel. In the first large-scale trials with GP IIb/IIIa inhibitors, abciximab was investigated. In aggregate, synthetic GP IIb/IIIa inhibitors, combined with aspirin and heparin, were shown to reduce ischemic events in patients with high- and low-risk coronary intervention, stents, unstable angina, and non-Q-wave infarction. With short-term use of synthetic GP IIb/IIIa inhibitors, there is no suppression of clinical evident restenosis 6 months after the end of treatment. With the doses currently used, bleeding occurs more often with the synthetic GP IIb/IIIa inhibitors (used for 3 days) than with abciximab (used for 12 hours), but there are no direct comparisons between these drugs. PMID- 10673267 TI - Gene therapy at a crossroads. PMID- 10673266 TI - Polymorphic ventricular tachycardia and repolarization abnormalities accompanying intracerebral hemorrhage. PMID- 10673268 TI - Genome race heats up. PMID- 10673269 TI - Smallpox virus gets another reprieve. PMID- 10673270 TI - State of health in State of the Union. PMID- 10673272 TI - First glimpses PMID- 10673271 TI - Monday a bad day for hearts. PMID- 10673273 TI - PON1 status and neurologic symptom complexes in Gulf War veterans. PMID- 10673274 TI - Genes for cognitive function: developments on the X. AB - Developments in human genome research enabled the first steps toward a molecular understanding of cognitive function. That there are numerous genes on the X chromosome affecting intelligence at the lower end of the cognitive range is no longer in doubt. Naturally occurring mutations have so far led to the identification of seven genes accounting for a small proportion of familial nonspecific X-linked mental retardation. These new data indicate that normal expression of many more X-linked and autosomal genes contribute to cognitive function. The emerging knowledge implicating genes in intracellular signaling pathways provides the insight to identify as candidates other X-linked and autosomal genes regulating the normal development of cognitive function. Recent advances in unravelling the underlying molecular complexity have been spectacular but represent only the beginning, and new technologies will need to be introduced to complete the picture. PMID- 10673275 TI - The genomic region encompassing the nephropathic cystinosis gene (CTNS): complete sequencing of a 200-kb segment and discovery of a novel gene within the common cystinosis-causing deletion. AB - Nephropathic cystinosis is an autosomal recessive disorder caused by the defective transport of cystine out of lysosomes. Recently, the causative gene (CTNS) was identified and presumed to encode an integral membrane protein called cystinosin. Many of the disease-associated mutations in CTNS are deletions, including one >55 kb in size that represents the most common cystinosis allele encountered to date. In an effort to determine the precise genomic organization of CTNS and to gain sequence-based insight about the DNA within and flanking cystinosis-associated deletions, we mapped and sequenced the region of human chromosome 17p13 encompassing CTNS. Specifically, a bacterial artificial chromosome (BAC)-based physical map spanning CTNS was constructed by sequence tagged site (STS)-content mapping. The resulting BAC contig provided the relative order of 43 STSs. Two overlapping BACs, which together contain all of the CTNS exons as well as extensive amounts of flanking DNA, were selected and subjected to shotgun sequencing. A total of 200,237 bp of contiguous, high-accuracy sequence was generated. Analysis of the resulting data revealed a number of interesting features about this genomic region, including the long-range organization of CTNS, insight about the breakpoints and intervening DNA associated with the common cystinosis-causing deletion, and structural information about five genes neighboring CTNS (human ortholog of rat vanilloid receptor subtype 1 gene, CARKL, TIP-1, P2X5, and HUMINAE). In particular, sequence analysis detected the presence of a novel gene (CARKL) residing within the most common cystinosis-causing deletion. This gene encodes a previously unknown protein that is predicted to function as a carbohydrate kinase. Interestingly, both CTNS and CARKL are absent in nearly half of all cystinosis patients (i.e., those homozygous for the common deletion). [The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AF168787 and AF163573.] PMID- 10673276 TI - Multiple LTR-retrotransposon families in the asexual yeast Candida albicans. AB - We have begun a characterization of the long terminal repeat (LTR) retrotransposons in the asexual yeast Candida albicans. A database of assembled C. albicans genomic sequence at Stanford University, which represents 14.9 Mb of the 16-Mb haploid genome, was screened and >350 distinct retrotransposon insertions were identified. The majority of these insertions represent previously unrecognized retrotransposons. The various elements were classified into 34 distinct families, each family being similar, in terms of the range of sequences that it represents, to a typical Ty element family of the related yeast Saccharomyces cerevisiae. These C. albicans retrotransposon families are generally of low copy number and vary widely in coding capacity. For only three families, was a full-length and apparently intact retrotransposon identified. For many families, only solo LTRs and LTR fragments remain. Several families of highly degenerate elements appear to be still capable of transposition, presumably via trans-activation. The overall structure of the retrotransposon population in C. albicans differs considerably from that of S. cerevisiae. In that species, retrotransposon insertions can be assigned to just five families. Most of these families still retain functional examples, and they generally appear at higher copy numbers than the C. albicans families. The possibility that these differences between the two species are attributable to the nonstandard genetic code of C. albicans or the asexual nature of its genome is discussed. A region rich in retrotransposon fragments, that lies adjacent to many of the CARE 2/Rel-2 sub-telomeric repeats, and which appears to have arisen through multiple rounds of duplication and recombination, is also described. PMID- 10673277 TI - The large srh family of chemoreceptor genes in Caenorhabditis nematodes reveals processes of genome evolution involving large duplications and deletions and intron gains and losses. AB - The srh family of chemoreceptors in the nematode Caenorhabditis elegans is very large, containing 214 genes and 90 pseudogenes. It is related to the str, stl, and srd families of seven-transmembrane or serpentine receptors. Like these three families, most srh genes are concentrated on chromosome V, and mapping of their chromosomal locations on a phylogenetic tree reveals 27 different movements of genes to other chromosomes. Mapping of intron gains and losses onto the phylogenetic tree reveals that the last common ancestral gene of the family had five introns, which are inferred to have been lost 70 times independently during evolution of the family. In addition, seven intron gains are revealed, three of which are fairly recent. Comparisons with 20 family members in the C. briggsae genome confirms these patterns, including two intron losses in C. briggsae since the species split. There are 14 clear C. elegans orthologs for these 20 genes, whose average amino acid divergence of 68% allows estimation of 85 gene duplications in the C. elegans lineage since the species split. The absence of six orthologs in C. elegans also indicates that gene loss occurs; consideration of all deletions and terminal truncations of srh pseudogenes reveals that large deletions are common. Together these observations provide insight into the evolutionary dynamics of this compact animal genome. PMID- 10673278 TI - Functional classification of cNMP-binding proteins and nucleotide cyclases with implications for novel regulatory pathways in Mycobacterium tuberculosis. AB - We have analyzed the cyclic nucleotide (cNMP)-binding protein and nucleotide cyclase superfamilies using Bayesian computational methods of protein family identification and classification. In addition to the known cNMP-binding proteins (cNMP-dependent kinases, cNMP-gated channels, cAMP-guanine nucleotide exchange factors, and bacterial cAMP-dependent transcription factors), new functional groups of cNMP-binding proteins were identified, including putative ABC transporter subunits, translocases, and esterases. Classification of the nucleotide cyclases revealed subtle differences in sequence conservation of the active site that distinguish the five classes of cyclases: the multicellular eukaryotic adenylyl cyclases, the eukaryotic receptor-type guanylyl cyclases, the eukaryotic soluble guanylyl cyclases, the unicellular eukaryotic and prokaryotic adenylyl cyclases, and the putative prokaryotic guanylyl cyclases. Phylogenetic distribution of the cNMP-binding proteins and cyclases was analyzed, with particular attention to the 22 complete archaeal and eubacterial genome sequences. Mycobacterium tuberculosis H37Rv and Synechocystis PCC6803 were each found to encode several more putative cNMP-binding proteins than other prokaryotes; many of these proteins are of unknown function. M. tuberculosis also encodes several more putative nucleotide cyclases than other prokaryotic species. PMID- 10673279 TI - Extensive genome-wide linkage disequilibrium in cattle. AB - A genome-wide linkage disequilibrium (LD) map was generated using microsatellite genotypes (284 autosomal microsatellite loci) of 581 gametes sampled from the dutch black-and-white dairy cattle population. LD was measured between all marker pairs, both syntenic and nonsyntenic. Analysis of syntenic pairs revealed surprisingly high levels of LD that, although more pronounced for closely linked marker pairs, extended over several tens of centimorgan. In addition, significant gametic associations were also shown to be very common between nonsyntenic loci. Simulations using the known genealogies of the studied sample indicate that random drift alone is likely to account for most of the observed disequilibrium. No clear evidence was obtained for a direct effect of selection ("Bulmer effect"). The observation of long range disequilibrium between syntenic loci using low-density marker maps indicates that LD mapping has the potential to be very effective in livestock populations. The frequent occurrence of gametic associations between nonsyntenic loci, however, encourages the combined use of linkage and linkage disequilibrium methods to avoid false positive results when mapping genes in livestock. PMID- 10673280 TI - Thermophilic bacteria strictly obey Szybalski's transcription direction rule and politely purine-load RNAs with both adenine and guanine. AB - When transcription is to the right of the promoter, the "top," mRNA-synonymous strand of DNA tends to be purine-rich. When transcription is to the left of the promoter, the top, mRNA-template strand tends to be pyrimidine-rich. This transcription-direction rule suggests that there has been an evolutionary selection pressure for the purine-loading of RNAs. The politeness hypothesis states that purine-loading prevents distracting RNA-RNA interactions and excessive formation of double-stranded RNA, which might trigger various intracellular alarms. Because RNA-RNA interactions have a distinct entropy-driven component, the pressure for the evolution of purine-loading might be greater in organisms living at high temperatures. In support of this, we find that Chargaff differences (a measure of purine-loading) are greater in thermophiles than in nonthermophiles and extend to both purine bases. In thermophiles the pressure to purine-load affects codon choice, indicating that some features of their amino acid composition (e.g., high levels of glutamic acid) might reflect purine loading pressure (i.e., constraints on mRNA) rather than direct constraints on protein structure and function. PMID- 10673281 TI - Genetic profile of insertion mutations in mouse leukemias and lymphomas. AB - Murine leukemia retroviruses (MuLVs) cause leukemia and lymphoma in susceptible strains of mice as a result of insertional mutation of cellular proto-oncogenes or tumor suppressor genes. Using a novel approach to amplify and sequence viral insertion sites, we have sequenced >200 viral insertion sites from which we identify >35 genes altered by viral insertion in four AKXD mouse strains. The class of genes most frequently altered are transcription factors, however, insertions are found near genes involved in signal transduction, cell cycle control, DNA repair, cell division, hematopoietic differentiation, and near many ESTs and novel loci. Many of these mutations identify genes that have not been implicated in cancer. By isolating nearly all the somatic viral insertion mutations contributing to disease in these strains we show that each AKXD strain displays a unique mutation profile, suggesting strain-specific susceptibility to mutations in particular genetic pathways. PMID- 10673282 TI - High-resolution physical map and transcript identification of a prostate cancer deletion interval on 8p22. AB - A genomic interval of approximately 1-1.5 Mb centered at the MSR marker on 8p22 has emerged as a possible site for a tumor suppressor gene, based on high rates of allele loss and the presence of a homozygous deletion found in metastatic prostate cancer. The objective of this study was to prepare a bacterial contig of this interval, integrate the contig with radiation hybrid (RH) databases, and use these resources to identify transcription units that might represent the candidate tumor suppressor genes. Here we present a complete bacterial contig across the interval, which was assembled using 22 published and 17 newly originated STSs. The physical map provides twofold or greater coverage over much of the interval, including 17 BACs, 15 P1s, 2 cosmids, and 1 PAC clone. The position of the selected markers across the interval in relation to the other markers on the larger chromosomal scale was confirmed by RH mapping using the Stanford G3 RH panel. Transcribed units within the deletion region were identified by exon amplification, searching of the Human Transcript Map, placement of unmapped expressed sequence tags (ESTs) from the Radiation Hybrid Database (RHdb), and from other published sources, resulting in the isolation of six unique expressed sequences. The transcript map of the deletion interval now includes two known genes (MSR and N33) and six novel ESTs. PMID- 10673283 TI - High-throughput SNP allele-frequency determination in pooled DNA samples by kinetic PCR. AB - We have developed an accurate, yet inexpensive and high-throughput, method for determining the allele frequency of biallelic polymorphisms in pools of DNA samples. The assay combines kinetic (real-time quantitative) PCR with allele specific amplification and requires no post-PCR processing. The relative amounts of each allele in a sample are quantified. This is performed by dividing equal aliquots of the pooled DNA between two separate PCR reactions, each of which contains a primer pair specific to one or the other allelic SNP variant. For pools with equal amounts of the two alleles, the two amplifications should reach a detectable level of fluorescence at the same cycle number. For pools that contain unequal ratios of the two alleles, the difference in cycle number between the two amplification reactions can be used to calculate the relative allele amounts. We demonstrate the accuracy and reliability of the assay on samples with known predetermined SNP allele frequencies from 5% to 95%, including pools of both human and mouse DNAs using eight different SNPs altogether. The accuracy of measuring known allele frequencies is very high, with the strength of correlation between measured and known frequencies having an r(2) = 0.997. The loss of sensitivity as a result of measurement error is typically minimal, compared with that due to sampling error alone, for population samples up to 1000. We believe that by providing a means for SNP genotyping up to thousands of samples simultaneously, inexpensively, and reproducibly, this method is a powerful strategy for detecting meaningful polymorphic differences in candidate gene association studies and genome-wide linkage disequilibrium scans. PMID- 10673284 TI - Population screening for haemochromatosis. PMID- 10673285 TI - Microsatellite instability in colitis associated colorectal cancer. PMID- 10673286 TI - How should endoscopic accessories be selected: trial or error? PMID- 10673287 TI - Combination of interferon alpha therapy with non-steroidal anti-inflammatory drugs in chronic hepatitis C. PMID- 10673288 TI - Bile acids are physiological ligands for a nuclear receptor. AB - Bile acids are essential for the solubilization and transport of dietary lipids and are the major products of cholesterol catabolism. Results presented here show that bile acids are physiological ligands for the farnesoid X receptor (FXR), an orphan nuclear receptor. When bound to bile acids, FXR repressed transcription of the gene encoding cholesterol 7 alpha-hydroxylase, which is the rate-limiting enzyme in bile acid synthesis, and activated the gene encoding intestinal bile acid-binding protein, which is a candidate bile acid transporter. These results demonstrate a mechanism by which bile acids transcriptionally regulate their biosynthesis and enterohepatic transport. PMID- 10673289 TI - Risk of ulcer bleeding in patients infected with Helicobacter pylori taking non steroidal anti-inflammatory drugs. AB - OBJECTIVE: To determine whether Helicobacter pylori is an independent risk factor for bleeding peptic ulcer in users of non-steroidal anti-inflammatory drugs (NSAIDs), including aspirin. DESIGN: A prospective matched case-control study. SETTING: Odense University Hospital, Denmark. SUBJECTS: 132 patients with a bleeding peptic ulcer (n=124) or haemorrhagic gastritis (n=8) at endoscopy who had taken an NSAID in the previous week and 136 controls who had taken NSAIDs without gastrointestinal complications. The controls were recruited from rheumatology and geriatric outpatient clinics. MEASUREMENTS: H pylori status assessed by serology and 13C-urea breath test and regarded as positive if either test was positive. Data on potential confounding factors including smoking and alcohol were collected by interview. MAIN RESULT: H pylori was present in 57% of cases and 43% of controls. The adjusted odds ratio of bleeding from a peptic ulcer owing to H pylori infection in NSAID users was 1.81 (95% CI 1.02 to 3.21) and was similar in aspirin and non-aspirin NSAID users. Peptic ulcer bleeding was also statistically significantly associated with a history of previous ulcer bleeding, dyspepsia within the previous 3 months, drinking alcohol but not with smoking. About 16% of bleeding peptic ulcers in NSAID users could be attributed to H pylori infection. CONCLUSION: NSAID users infected with H pylori have an almost doubled risk of bleeding peptic ulcer compared with uninfected NSAID users. PMID- 10673290 TI - The human trefoil peptide, TFF1, is present in different molecular forms that are intimately associated with mucus in normal stomach. AB - BACKGROUND: TFF1 is a 6.5 kDa secreted protein that is expressed predominantly in normal gastric mucosa. It is coexpressed with mucins and it can form dimers via a free carboxy terminal cysteine residue. AIMS: To investigate the molecular forms of TFF1 that are present in normal human stomach and the association of the different molecular forms with mucus. SUBJECTS: All subjects had macroscopically normal stomachs at gastroscopy. None had a significant past medical history. METHODS: TFF1 was detected in normal gastric mucosa and adherent mucus by western transfer analysis after electrophoresis on reducing and non-reducing polyacrylamide gels. In some instances, proteins were fractionated by caesium chloride density gradient centrifugation prior to detection of TFF1. The location of TFF1 in gastric mucosa with an intact adherent mucus layer was assessed by immunohistochemistry. RESULTS: Three different molecular forms of TFF1 were detected: TFF1 monomer, TFF1 dimer, and a TFF1 complex with an apparent molecular mass of about 25 kDa. TFF1 was present at higher concentrations than realised previously. The TFF1 complex was present in the adherent mucus gel layer but while its interaction with mucin was destabilised by caesium chloride, the interaction between mucin and the TFF1 dimer was resistant to caesium chloride. CONCLUSIONS: Most of TFF1 in normal human gastric mucosa is present in a complex that is stabilised by a disulphide bond. TFF1 is intimately associated with mucus. The high concentration, colocalisation, and binding of TFF1 to gastric mucus strongly implicate TFF1 in gastric mucus function. PMID- 10673291 TI - Importance of Helicobacter pylori cagA and vacA status for the efficacy of antibiotic treatment. AB - BACKGROUND: Virulence factors of Helicobacter pylori are associated with peptic ulcer disease and may be also associated with the efficacy of treatment. AIMS: To determine the relation between the vacA and the cagA status of H pylori, clinical disease, and treatment outcome. PATIENTS: 121 patients with H pylori infection and peptic ulcer disease or functional dyspepsia were treated by quadruple antibiotic therapy in two groups for one and two days, respectively. METHODS: DNA was isolated from gastric antral biopsy specimens, taken before and after treatment, and the vacA and cagA status was determined by polymerase chain reaction and reverse hybridisation. RESULTS: Peptic ulcer disease was significantly associated with the vacA s1 type, and cagA positivity, but not with the vacA m type. Treatment efficacy was significantly higher in patients with peptic ulcer disease, or infected with cagA+/vacA s1 strains. CONCLUSIONS: The strong association between the cagA and vacA status and peptic ulcer disease was confirmed. Cure rates seem to be higher for patients with cagA+/vacA s1 H pylori strains, which is consistent with the higher cure rate observed among ulcer patients compared with functional dyspepsia patients. Therefore, treatment studies may require stratification for presence of ulcers as well as H pylori genotypes. PMID- 10673292 TI - Lack of cellular and humoral immunological responses to oats in adults with coeliac disease. AB - OBJECTIVE: Recent research suggests that oats do not harm intestinal villi in adults with coeliac disease. As the immunological effects of oats have not been examined in detail, it was decided to compare the immunological responses of a gluten free diet including oats with those of a conventional gluten free diet. DESIGN: A randomised controlled intervention study over 6-12 months. SUBJECTS: Forty adults with newly diagnosed coeliac disease and 52 with coeliac disease in remission were examined. INTERVENTION: The effects of a gluten free diet including oats and a conventional gluten free diet were compared. MAIN OUTCOME MEASURES: Serum levels of gliadin and reticulin antibodies as well as numbers of intraepithelial lymphocytes (IELs) in intestinal mucosa were examined before and after the intervention. RESULTS: The rate of disappearance of gliadin and reticulin antibodies did not differ between the diet groups in patients with newly diagnosed coeliac disease. Oats also had no effect on gliadin or reticulin antibody levels in the patients with remission. The number of IELs decreased similarly regardless of the diet of newly diagnosed patients, and no increase in the number of IELs was found in the patients in remission with or without oats. CONCLUSIONS: These results strengthen the view that adult patients with coeliac disease can consume moderate amounts of oats without adverse immunological effects. PMID- 10673293 TI - Coeliac disease and unfavourable outcome of pregnancy. AB - BACKGROUND: Up to 50% of women with untreated coeliac disease experience miscarriage or an unfavourable outcome of pregnancy. In most cases, after 6-12 months of a gluten free diet, no excess of unfavourable outcome of pregnancy is observed. The prevalence of undiagnosed coeliac disease among pregnant women is not known. AIM: To determine the prevalence of untreated coeliac disease among women attending the obstetrics-gynaecological department. METHODS: Endomysial antibodies, which are specific and sensitive for coeliac disease, were evaluated in all women attending the obstetrics-gynaecology department of a large city hospital over a 90 day period. RESULTS: Of 845 pregnant women screened, 12 were identified as having coeliac disease. Three had previously been diagnosed but were not following a gluten free diet. The remaining nine underwent a small intestinal biopsy, which confirmed the diagnosis. The outcome of pregnancy was unfavourable in seven of these 12 women. Six healthy babies were born with no problems after the women had been on a gluten free diet for one year. CONCLUSIONS: Overall, 1 in 70 women was affected by coeliac disease, either not diagnosed (nine cases) or not treated (three cases). Their history of miscarriages, anaemia, low birth weight babies, and unfavourable outcome of pregnancy suggests that testing for coeliac disease should be included in the battery of tests prescribed for pregnant women. Coeliac disease is considerably more common than most of the diseases for which pregnant women are routinely screened. Unfavourable events associated with coeliac disease may be prevented by a gluten free diet. PMID- 10673294 TI - Ulcerative colitis in Olmsted County, Minnesota, 1940-1993: incidence, prevalence, and survival. AB - BACKGROUND: There is significant geographic variation in the reported incidence of ulcerative colitis. AIMS: To update the incidence and prevalence of ulcerative colitis in Olmsted County, Minnesota, examine temporal trends, and determine overall survival. PATIENTS: All Olmsted County residents diagnosed with ulcerative colitis between 1940 and 1993 (incidence cases), and all residents with ulcerative colitis alive on 1 January 1991 (prevalence cases). METHODS: Incidence and prevalence rates were adjusted using 1990 US census figures for whites. The effects of age, sex, and calendar year on incidence rates were evaluated using Poisson regression. Survival from diagnosis was compared with that expected for US north-central whites. RESULTS: Between 1940 and 1993, 278 incidence cases were identified, for an adjusted incidence rate of 7.6 cases per 100 000 person years (95% confidence interval (CI), 6.7 to 8.5). On 1 January 1991, there were 218 residents with definite or probable ulcerative colitis, for an adjusted prevalence rate of 229 cases per 100 000 (95% CI, 198 to 260). Increased incidence rates were associated with later calendar years (p<0.002), younger age (p<0.0001), urban residence (p<0.0001), and male sex (p<0.003). Overall survival was similar to that expected (p>0.2). CONCLUSIONS: The overall incidence rate of ulcerative colitis in Olmsted County increased until the 1970s, and remained stable thereafter. Incidence rates among men and urban residents were significantly higher. The prevalence rate in Rochester in 1991 was 19% higher than that in 1980. Overall survival was similar to that of the general population. PMID- 10673295 TI - Interleukin 10 gene transfer prevents experimental colitis in rats. AB - BACKGROUND: The development of colitis in interleukin 10 (IL-10) deficient mice, together with the known anti-inflammatory and immunomodulatory properties of this cytokine have prompted consideration of IL-10 as a treatment for inflammatory bowel disease (IBD). However, studies using hrIL-10 in IBD models have yielded inconsistent results. AIMS: To examine the therapeutic potential of overexpressing the IL-10 gene before and after the induction of experimental colitis in rats. METHODS: Gene transfer was achieved by intraperitoneal injection of non-replicating human type 5 adenovirus bearing the IL-10 gene, either 24 hours before or one hour after intrarectal administration of dinitrobenzene sulphonic acid in rats. Colonic damage and inflammation was assessed macroscopically and by measuring myeloperoxidase activity and leukotriene B4 concentrations. RESULTS: Gene transfer increased IL-10 protein in serum for up to six days. IL-10 gene transfer prior to colitis improved colitis macroscopically and histologically, and significantly reduced colonic myeloperoxidase activity and leukotriene B4 concentrations. In contrast, IL-10 gene transfer after the onset of colitis had no beneficial effect. CONCLUSIONS: Gene therapy using an adenovirus-IL-10 construct was successful in preventing but not in reversing experimental colitis in the rat. PMID- 10673296 TI - Human intestinal epithelial cells secrete interleukin-1 receptor antagonist and interleukin-8 but not interleukin-1 or interleukin-6. AB - BACKGROUND: There is growing evidence that intestinal epithelial cells (IECs) are involved in the mucosal immune system. AIM: To assess the pattern of cytokines secreted by IECs and lamina propria mononuclear cells (LPMNCs). To achieve this, the expression and secretion of interleukin (IL)-1, IL-1 receptor antagonist (IL 1ra), IL-6, and IL-8 in human primary colonic and ileal IECs and LPMNCs from the same patient were studied. METHODS: IECs and LPMNCs were isolated from surgical specimens or endoscopic biopsy samples. mRNA expression was investigated by northern blot analysis. Secretion of IL-1beta, IL-6, IL-8, and IL-1ra was measured by enzyme linked immunosorbent assay. RESULTS: IL-1ra mRNA levels were higher in IECs than in LPMNCs in all probands. IL-8 mRNA was only present in low amounts in the IECs from two controls. In none of the specimens were IL-1beta and IL-6 mRNA present in IECs. Transcripts encoding IL-1beta, IL-1ra, IL-6, and IL-8 were identified in LPMNC preparations of all specimens. IECs from normal mucosa produced no detectable amounts of IL-1beta or IL-6, whereas LPMNCs did. IECs secreted some IL-8 (65 (9) pg/10(5) cells) but significantly more was generated by LPMNCs (408 (43) pg/10(5) cells, p<0.0001). However, IECs secreted more IL-1ra than did LPMNCs (120 (12) v 94 (11) pg/10(5) cells). In acute inflammation, IEC IL-1ra secretion was significantly increased. A correlation between secreted IL 1ra and the macroscopical degree of inflammation was found in Crohn's disease (r = 0.64, p<0.0001, n = 36) and ulcerative colitis (r = 0. 76, p<0.0001, n = 24). CONCLUSIONS: IECs from normal mucosa express and secrete IL-1ra and low amounts of IL-8, but no IL-1 or IL-6. In inflamed mucosa the secretion of IL-1ra by IECs is slightly increased but may be not sufficient to antagonise the greatly increased production of proinflammatory cytokines by LPMNCs and the IECs themselves. PMID- 10673297 TI - Altered mRNA expression of glycosyltransferases in human colorectal carcinomas and liver metastases. AB - BACKGROUND/AIMS: Biosynthesis of carbohydrate structures is tissue specific and developmentally regulated by glycosyltransferases such as fucosyltransferases, sialyltransferases, and N-acetylgluco- saminyltransferases. During carcinogenesis, aberrant glycosylation leads to the development of tumour subpopulations with different adhesion properties. Therefore alterations in glycosyltransferase mRNA expression in colorectal carcinomas were examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). METHODS: Colorectal carcinoma specimens were classified and characterised according to the WHO/UICC system. Expression of fucosyltransferases FT-I, FT-III, FT-IV, FT-V, FT-VI, and FT-VII, sialyltransferases ST3Gal-I, ST3Gal-III, ST3Gal IV, and ST6Gal-I, beta1,4-galacto- syltransferase, and beta1,6-Nacetylgluco- saminyltransferase V (GNT-V) was screened simultaneously in extracts of 22 homogenised tumour specimens by RT-PCR and compared with corresponding mucosa from each patient. Also 12 adenomas and 17 liver metastases of colorectal carcinomas were examined. RESULTS: GNT-V expression was enhanced in colorectal adenomas (p = 0.039), carcinomas (p<0.001), and liver metastases of colorectal carcinomas (p<0.001). Also, expression of fucosyltransferase FT-IV was increased in colorectal adenomas (p = 0.039) and carcinomas (p<0. 001). In addition, fucosyltransferase FT-I (p<0.001) and sialyltransferases ST6Gal-I (p = 0.004) and ST3Gal-III (p = 0.001) showed increased expression in carcinoma specimens. On the other hand, fucosyltransferase FT-III was less abundantly expressed in carcinomas exhibiting distant metastases (p = 0.046) and in highly invasive tumours (p = 0.041). CONCLUSIONS: Glycosyltransferase mRNA expression is significantly altered in colorectal adenomas and carcinomas isolated from surgical specimens. RT-PCR determination of specific glycosyltransferases may be helpful for earlier detection of carcinomas and for tumour prognosis. PMID- 10673299 TI - Pathways and receptors involved in peptide YY induced contraction of rat proximal colonic muscle in vitro. AB - BACKGROUND: Peptide YY (PYY) is involved in the regulation of several gut functions, including secretion and motility. It exerts its effects through a family of six receptors, commonly named the Y receptor family. AIMS: To characterise the effects of PYY on strips of rat proximal colon in vitro, and to determine the pathways and receptors involved. METHODS: Contractions of strips removed from the muscle layer of rat proximal colon were recorded under isometric conditions, using PYY, Y receptor agonists and antagonists, and nerve blockers. Reverse transcription-polymerase chain reaction was also performed to detect the presence of mRNA coding for Y receptors. Finally, smooth muscle cells were isolated to estimate the cell length and intracellular Ca(2+) concentration in the presence and absence of PYY. RESULTS: PYY, neuropeptide Y (NPY), pancreatic polypeptide (PP) and [Leu31,Pro34]NPY induced a dose dependent contraction of strips from proximal colon. Tetrodotoxin partially inhibited the PYY and NPY induced contractions, and strongly inhibited the PP induced contraction. Specific antagonists showed the involvement of cholinergic nicotinic receptors and NK1 receptor. BIBP 3226, a specific Y1 antagonist, did not modify the colonic smooth muscle response to PYY, whereas blocking L-type Ca(2+) channels with D-600 abolished its effects. Moreover, PYY induced an increase in intracellular Ca(2+) concentration, associated with a reduction in cell length. mRNA encoding Y1 and Y4 receptors were detected in the muscle strips. CONCLUSIONS: These findings suggest that PYY stimulates colonic contractile activity in vitro through (a) a nervous Y4 dependent pathway and (b) a pathway involving a potential new receptor on myocytes. PMID- 10673298 TI - Defective hMSH2/hMLH1 protein expression is seen infrequently in ulcerative colitis associated colorectal cancers. AB - BACKGROUND: Ulcerative colitis is associated with an increased risk of colorectal cancer above that of the normal population. The relative risk correlates with the extent and duration of the disease but the genetic basis of ulcerative colitis associated cancer risk is not known. AIMS: To assess the prevalence of microsatellite instability and mismatch repair gene abnormalities in ulcerative colitis associated colorectal cancer. PATIENTS: Forty six patients with colorectal cancer, with a previous histological diagnosis of ulcerative colitis. METHODS: The frequency of microsatellite instability and/or immunohistochemical expression of hMSH2 and hMLH1 was assessed. Thirty three cases were investigated using both approaches. RESULTS: Although 6/41 (14.6%) cases showed microsatellite instability at one or more markers, only one case (2. 4%) exhibited high level instability (at least two markers affected). Of 38 cases which were assessed using antibodies against hMSH2 and hMLH1, only one case (2.6%) showed loss of expression. This case, which showed loss of hMSH2 expression, was the same case which exhibited high level microsatellite instability. The 33 cases which were investigated using both approaches showed that loss of expression of either hMSH2 or hMLH1 was not seen in any case which exhibited microsatellite instability in no more than one marker. CONCLUSIONS: This study suggests that both high level microsatellite instability and loss of expression of hMSH2/hMLH1 are infrequent events in ulcerative colitis associated colorectal cancers. Low level microsatellite instability was not associated with loss of expression of either hMSH2 or hMLH1. PMID- 10673300 TI - Pressure and frequency dependent linkage between motility and epithelial secretion in human proximal small intestine. AB - BACKGROUND: Motor disturbances are sometimes associated with diarrhoea by unknown mechanisms. AIM: To determine if there is a quantitative link between intestinal motility and epithelial secretion. SUBJECTS: Experiments were performed in 21 healthy volunteers and three patients with villus atrophy. METHODS: Duodenal and jejunal motor activities were registered in the fasted state by open tip manometry. Secretion was measured directly by marker perfusion and indirectly by recording transmural potential difference (PD). RESULTS: A significant correlation was found between "low pass filtered" pressure and PD, but no correlation was found between amplitudes of isolated contractions and PD changes. During repeated phasic contractions (phase III of migrating motor complex), PD increased at a rate that was higher in the duodenum than in the jejunum, and higher in patients with villus atrophy than in healthy controls. After reaching a peak, PD decreased despite continuing phasic motor activity, provided that there was no concomitant increase in mean pressure. Fluid secretion increased roughly in parallel with PD, except at the very end of the cycle. CONCLUSIONS: To explain these findings, one has to postulate participation of at least two types of receptor: a slowly adapting pressure sensitive receptor and another mechanoreceptor, possibly a mucosal touch receptor, to account for the run down phenomenon. This model predicts that short lasting trains of contractions, so called discrete clusters, will be a particularly potent stimulus for activation of mucosal secretion. PMID- 10673301 TI - Acute pancreatitis in peritoneal dialysis and haemodialysis: risk, clinical course, outcome, and possible aetiology. AB - BACKGROUND: It has been suggested that the incidence of acute pancreatitis in patients with end stage renal failure is increased. AIMS: To assess the risk of acute pancreatitis in patients on long term peritoneal dialysis and long term haemodialysis compared with the general population, to evaluate its clinical course and outcome, and to identify possible aetiological factors. PATIENTS: All patients who were maintained on long term peritoneal dialysis and/or haemodialysis (total dialysis time more than six weeks) from January 1989 to March 1998 in a large general hospital in The Netherlands. METHODS: Retrospective cohort study. Standardised ratios (as an approximate relative risk) between the incidence of acute pancreatitis in haemodialysis or peritoneal dialysis and the general population were calculated. Possible risk factors were identified. Patients with and without acute pancreatitis were compared. RESULTS: In 269 patients on haemodialysis (total of 614 person years), one patient developed an attack of acute pancreatitis. Patients on haemodialysis did not show an increased risk for acute pancreatitis compared with the general population (standardised ratio 11; 95% confidence interval (CI) 0.275 to 60.5). In 128 patients on peritoneal dialysis (total of 241 person years), seven patients had nine attacks of acute pancreatitis. Patients on peritoneal dialysis had a significantly and highly increased risk for acute pancreatitis (standardised ratio 249; 95% CI 114 to 473). Mortality in this series of nine attacks was 11%. No single aetiological risk factor could be identified. CONCLUSIONS: The risk of acute pancreatitis in patients on long term peritoneal dialysis is significantly and highly increased compared with the general population. The underlying causal mechanisms remain to be elucidated. PMID- 10673302 TI - Effect of endothelin and endothelin receptor blockade on capillary permeability in experimental pancreatitis. AB - BACKGROUND: Capillary leakage with fluid loss into the third space contributes to many of the early systemic complications in severe acute pancreatitis. There has been increasing interest in endothelin as one of the factors affecting capillary permeability. AIM: To elucidate further the role of endothelin in the development of capillary leakage in acute pancreatitis by investigating the effect of exogenous endothelin administration and endothelin receptor blockade in sham operated animals and two models of acute pancreatitis. METHODS: Determination of capillary permeability in the pancreas and colonic mucosa by quantifying extravasation of fluorescein labelled dextran using a novel computer assisted video image analysis system. RESULTS: Pancreatic and colonic capillary permeability increased stepwise from mild to severe acute pancreatitis. Endothelin increased pancreatic and colonic capillary permeability in healthy animals and animals with mild acute pancreatitis but had no additional adverse effect in severe acute pancreatitis. Endothelin receptor blockade decreased pancreatic capillary permeability in sham operated rats but had no effect on the colon. In mild and severe acute pancreatitis, endothelin receptor blockade stabilised increased capillary permeability in both the pancreas and colon. CONCLUSIONS: Endothelin plays an important role in mediating capillary permeability in the pancreas. In severe pancreatitis, it increases capillary permeability even outside the pancreas, thereby contributing to capillary leakage. Endothelin receptor blockade significantly reduces capillary permeability in acute pancreatitis both in and outside the pancreas, suggesting a therapeutic approach to counteract capillary leakage in severe acute pancreatitis. PMID- 10673303 TI - A prospective randomised multicentre trial comparing 10 Fr Teflon Tannenbaum stents with 10 Fr polyethylene Cotton-Leung stents in patients with malignant common duct strictures. AB - BACKGROUND: Stent blockage is a multifactorial process in which stent design and materials, bacteria, proteins, and bile viscosity play a role. AIMS: To compare the patency of the 10 Fr Teflon Tannenbaum (TT) stent to that of the 10 Fr Cotton Leung (CL) polyethylene stent with sideholes, in patients with malignant obstructive jaundice. METHODS: Patients were recruited to this prospective multicentre randomised study if they had a newly diagnosed malignant bile duct stricture below the hilum of the liver suitable for stenting with a 10 Fr stent. Data were collected and monitored by a professional monitoring company. Primary patency was the interval between stent placement and first exchange or death without recurrent jaundice. RESULTS: 134 consecutive patients were recruited between November 1994 and June 1997; 65 were randomised to the TT stent and 69 to the CL stent. Median patency and 95% confidence intervals were 181 (59, 303) days for the TT stent and 133 (92, 174) days for the CL stent, with no significant difference between the two stents (p=0.49). Median survival and 95% confidence intervals were 115 (71, 159) days for the TT stent and 151 (112, 190) days for the CL stent, with no significant difference between the two stents (p=0.765). CONCLUSION: Neither Teflon as a stent material nor the Tannenbaum design prolong the patency of plastic stents. PMID- 10673304 TI - Incidence of liver disease in people with HFE mutations. AB - BACKGROUND: Most patients with haemochromatosis have mutations of the HFE gene. However, the risk to people with HFE mutations of developing disease manifestations of haemochromatosis is not known. AIMS: To determine the risk of developing cirrhosis and liver cancer in individuals with HFE mutations in a population where few people were being treated for haemochromatosis. METHODS: 215 archive biopsy specimens of liver cancer (n=34) and cirrhosis (n=190) were retrieved from histology archives. Blood samples from 1000 individuals from the normal population were also collected. DNA was extracted from the biopsy specimens and exons 2 and 4 of the HFE gene were amplified using polymerase chain reaction. The products were analysed for the C282Y (845A) and H63D (187G) mutations. RESULTS: Three (8.8%) patients from the liver cancer group were homozygous for the C282Y mutation. Five (2.6%) patients from the cirrhosis group were homozygous for the C282Y mutation. One case fell in both the liver cancer and cirrhosis groups. C282Y homozygosity was thus significantly more frequent in both groups than in the normal population. These 215 cases are representative of a population of about 250 000 over 20 years. During this period we estimate that about 260 births or deaths of C282Y homozygous individuals occurred within this population. CONCLUSIONS: A diagnosis of liver cancer or cirrhosis is rare in the lifetime of individuals from this population who are homozygous for the C282Y mutation (2.5%; upper 95% confidence interval (CI) = 8%). Similarly liver disease is rare among C282Y/H63D compound heterozygotes (1%; upper 95% CI = 3.5%). PMID- 10673305 TI - Automated measurement of unsaturated iron binding capacity is an effective screening strategy for C282Y homozygous haemochromatosis. AB - BACKGROUND: C282Y hereditary haemochromatosis is an appropriate condition for population screening. Transferrin saturation, the best screening test to date, is relatively expensive, labour intensive, and cannot be automated. Unsaturated iron binding capacity is a surrogate marker of transferrin saturation and its measurement can be automated. AIMS: To evaluate a screening strategy for C282Y hereditary haemochromatosis in a tertiary hospital environment based on unsaturated iron binding capacity as the initial screening test. METHODS: Measurement of unsaturated iron binding capacity was adapted to the main laboratory analyser. An unsaturated iron binding capacity of less than 30 micromol/l was identified as an appropriate decision point and 5182 consecutive subjects were screened over 28 consecutive days. RESULTS: Of those screened, 697 had an unsaturated iron binding capacity less than 30 micromol/l. Of these, transferrin saturation was greater than 40% in 294. A total of 227 were able to be genotyped for the C282Y mutation. Nine subjects homozygous for C282Y were identified. Based on full cost recovery, affected persons were identified at a cost of Aus$2268.77 per case (approximately US$1496). CONCLUSION: Automated measurement of unsaturated iron binding capacity enables a cost effective, large scale population screening programme for C282Y hereditary haemochromatosis to be developed. PMID- 10673306 TI - Serum antibodies to Helicobacter hepaticus and Helicobacter pylori in patients with chronic liver disease. AB - BACKGROUND: Bile tolerant helicobacter species such as H hepaticus and H bilis have frequently been reported to cause hepatitis in mice and other rodents. AIMS: To investigate the possible pathogenic role of these and other helicobacter species in chronic liver disease in humans. METHODS: Serum samples from 144 patients with various chronic liver diseases, 30 patients with primary sclerosing cholangitis (PSC), and 48 healthy blood donors were analysed for antibodies against H hepaticus murine strain CCUG 33637 and H pylori strain CCUG 17874. Cell surface proteins of H hepaticus were extracted by acid glycine buffer and used in an enzyme immunoassay (EIA) and immunoblot (IB). RESULTS: 56 of 144 (39%) patients with chronic liver diseases and six of 30 (20%) with PSC showed increased antibody concentrations in the H hepaticus EIA; in the H pylori EIA the numbers were 58% and 13% respectively. Compared with the healthy blood donors the antibody reactivity against the two helicobacter species was not increased (46% and 48% respectively). Patient serum samples retested by the H hepaticus EIA after absorption with sonicated H pylori cells remained positive in 12 of 37 (33%) serum samples. Distinct antibody reactivity to 55-65 kDa proteins was observed by H hepaticus IB, after the absorption step, and was considered specific for H hepaticus. These 12 serum samples were from patients with chronic alcoholic liver disease. CONCLUSIONS: Antibodies to H hepaticus, often cross reacting with H pylori, occur frequently in patients with chronic liver diseases, with no clear cut relation to specific diagnostic groups. The pathogenic significance of these findings is not known. PMID- 10673307 TI - Diagnosis of Wilson's disease: an experience over three decades. AB - BACKGROUND: Wilson's disease is a rare but treatable condition that often presents diagnostic dilemmas. These dilemmas have for the most part not been resolved by the identification and cloning of the Wilson's disease gene. AIMS: To report our experience over three decades with patients with Wilson's disease in order to illustrate the diverse patterns of presentation and thereby broaden the approach to diagnosis. METHODS: Clinical and laboratory findings of 30 patients with Wilson's disease were reviewed. RESULTS: Twenty two patients presented with liver manifestations (eight with fulminant hepatic failure and 14 with chronic liver disease), three with neurological disease, and one with haemolysis; four were asymptomatic siblings of patients with Wilson's disease. Seventy per cent were diagnosed within six months of the onset of symptoms, but diagnosis was delayed for up to nine years. Age range at diagnosis was wide (7-58 years) and five patients were over 40. In patients presenting with non-fulminant disease, 18% had neither Kayser-Fleischer rings nor low caeruloplasmin concentrations. Increased liver copper concentrations were found in all but one patient who had undergone six years of penicillamine treatment. In fulminant hepatic failure (n=8) additional features helpful in the diagnosis included evidence of haemolysis, increased urinary copper (range 844-9375 microg/24 h), and a high non caeruloplasmin copper (range 325-1743 microg/l). CONCLUSIONS: The diagnosis of Wilson's disease still depends primarily on the evaluation of clinical and laboratory evidence of abnormal copper metabolism. No one feature is reliable, but the diagnosis can usually be made provided that it is suspected. Wilson's disease should be considered in patients of any age with obscure hepatic or neurological abnormalities. PMID- 10673308 TI - Influence of hepatitis delta virus infection on morbidity and mortality in compensated cirrhosis type B. The European Concerted Action on Viral Hepatitis (Eurohep). AB - BACKGROUND: The effect of hepatitis delta virus (HDV) infection on the clinical course of cirrhosis type B is poorly defined. AIMS: To investigate the impact of HDV status on morbidity and mortality in cirrhosis type B. PATIENTS/METHODS: Retrospective cohort study of 200 Western European patients with compensated cirrhosis type B followed for a median period of 6.6 years. RESULTS: At diagnosis, 20% of patients had antibodies to HDV (anti-HDV); median age was lower in anti-HDV positive cirrhotics (34 v 48 years respectively). Kaplan-Meier five year probability of hepatocellular carcinoma (HCC) was 6, 10, and 9% in anti-HDV positive/HBeAg negative, anti-HDV negative/HBeAg negative, and anti-HDV negative/HBeAg positive cirrhotics respectively; the corresponding figures for decompensation were 22, 16, and 19% and for survival they were 92, 89, and 83% respectively. Cox regression analysis identified age, albumin concentration, gamma-globulin concentration, and HDV status as significant independent prognostic variables. After adjustment for clinical and serological differences at baseline, the risk (95% confidence interval) for HCC, decompensation, and mortality was increased by a factor of 3.2 (1.0 to 10), 2.2 (0.8 to 5.7), and 2.0 (0.7 to 5.7) respectively in anti-HDV positive relative to HDV negative cirrhotic patients. The adjusted estimated five year risk for HCC was 13, 4, and 2% for anti-HDV positive/HBeAg negative, anti-HDV negative/HBeAg negative, and anti-HDV negative/HBeAg positive cirrhotics respectively; the corresponding figures for decompensation were 18, 8, and 14% and for survival 90, 95, and 93% respectively. CONCLUSIONS: HDV infection increases the risk for HCC threefold and for mortality twofold in patients with cirrhosis type B. PMID- 10673309 TI - Interferon-alpha 2b combined with daily ketoprofen administration improves virological response in chronic hepatitis C: a prospective and randomised trial. AB - BACKGROUND: Less than 15% of patients with chronic hepatitis C show a sustained virological response to interferon treatment. AIM: To evaluate the efficacy and safety of different doses of ketoprofen combined with interferon-alpha 2b in the treatment of chronic hepatitis C. PATIENTS/METHODS: Seventy compensated patients with chronic hepatitis C received interferon-alpha 2b 3 million units three times a week for six months. They were randomly assigned to: group 1 (n = 23), interferon-alpha 2b alone; group 2 (n = 23), interferon-alpha 2b plus 200 mg ketoprofen three times a week; group 3 (n = 24), interferon-alpha 2b plus 200 mg ketoprofen twice a day. Complete and sustained responses were defined as normal serum alanine aminotransferase levels and negative serum hepatitis C virus RNA at six and 12 months respectively. RESULTS: Complete and sustained responses were similar in groups 1 and 2: 10% v 5% and 5% v 0% respectively. In group 3, complete response was 29% (p = 0.13 v group 1 and p = 0.04 v group 2) and sustained response was 26% (p = 0.07 v group 1 and p = 0.01 v group 2). Overall, adverse events were similar in the three groups. However, 'flu-like syndrome was less common in group 2 (30%) and group 3 (37%) than in group 1 (77%) (p = 0.01). CONCLUSIONS: Twice daily ketoprofen administration combined with interferon-alpha 2b produced an increase in complete and sustained responses. Although the combination of interferon-alpha 2b with ketoprofen was well tolerated and decreased the incidence of 'flu-like syndrome, it is advisable to monitor possible non-steroid anti-inflammatory drug hepatotoxicity. PMID- 10673310 TI - Possible paracrine growth of adenocarcinoma of the stomach induced by granulocyte colony stimulating factor produced by squamous cell carcinoma of the oesophagus. AB - Synchronous cancers of the oesophagus and stomach diagnosed in a patient showing pronounced leucocytosis were examined for production of granulocyte colony stimulating factor (G-CSF) and expression of G-CSF receptor. Whereas enzyme immunoassay of tissue extracts showed that the oesophageal carcinoma produced G CSF, the gastric cancer did not. However, the gastric tumour showed G-CSF receptor expression on immunohistochemical examination of sections. These findings suggest that the oesophageal cancer promoted gastric cancer growth by paracrine mechanisms involving G-CSF. PMID- 10673311 TI - Lidocaine toxicity in a student undergoing upper gastrointestinal endoscopy. PMID- 10673312 TI - Peroxynitrite and inflammatory bowel disease. PMID- 10673314 TI - Molecule liftoff from surfaces. AB - Molecular dynamics simulations have been used to model the kiloelectronvolt particle bombardment of organic layers on metal substrates such as occurs in the analytical techniques of secondary ion mass spectrometry and fast atom bombardment mass spectrometry. Vignettes of insights gained from the simulations along with comparisons to experimental data are presented in this Account. Topics include intact molecular ejection vs fragmentation, prediction of reaction pathways, influence of the substrate, and quantitative predictions of energy and angular distributions. PMID- 10673315 TI - Hybrid electrochemical/chemical synthesis of quantum dots. AB - The "electrochemical/chemical method" (or "E/C method") is a new wet chemical method for synthesizing semiconductor quantum dots on graphite surfaces. The E/C synthesis of quantum dots composed of the generic semiconducting salt, MX, typically involves three steps: (1) electrochemical deposition of nanoparticles of the metal, M degrees, from a solution of metal ions, M(n)(+); (2) electrochemical oxidation of these metal particles to MO(n)()(/2), and; (3) displacement of the oxygen from MO(n)()(/2) using HX (for example) to yield nanoparticles of MX. This conversion from metal to metal oxide to metal salt occurs on a particle-by-particle basis; that is, each metal nanoparticle is converted into a semiconductor nanoparticle. E/C-synthesized beta-CuI and CdS quantum dots possess many of the attributes of quantum dots synthesized using molecular beam epitaxy, including epitaxial orientation on the graphite surface, a narrow size dispersion, and strong, particle size-tunable photoluminescence. However, the E/C method is faster, cheaper, and applicable to a greater number of materials. PMID- 10673316 TI - What controls the rates of interprotein electron-transfer reactions. AB - Rates of electron-transfer (ET) reactions are dependent on driving force, reorganizational energy, distance, and the nature of the medium which the electron must traverse. In kinetically complex biological systems, non-ET reactions may be required to activate the system for ET and may also influence the observed rates. Studies of ET from tryptophan tryptophylquinone to copper to heme in the methylamine dehydrogenase-amicyanin-cytochrome c-551i ET complex, as well as studies of other physiologic redox protein complexes, are used to illustrate the combination of factors which control rates of interprotein ET reactions. PMID- 10673317 TI - Biomaterials in drug delivery and tissue engineering: one laboratory's experience. AB - This Account reviews our laboratory's research in biomaterials. In one area, drug delivery, we discuss the development of materials that are capable of releasing macromolecules such as proteins and peptides, intelligent delivery systems based on magnetism or microchip technology, new degradable materials such as polyanhydrides, and noninvasive approaches for delivering molecules through the skin and lungs. A second area, tissue engineering, is also discussed. New polymer systems for creating cartilage, blood vessels, nerves, and other tissues are examined. PMID- 10673318 TI - Organic synthesis and chemical ecology. AB - The roles of organic synthesis in chemical ecology are discussed, with many examples. The structure, including the absolute configuration, of a semiochemical (signal substance) can be established by enantioselective synthesis. Only through synthesis can a semiochemical be obtained in an amount sufficient for decisive biological evaluation. Rigorous enantioselective synthesis of semiochemicals to provide their pure enantiomers has shown that they are not always enantiomerically pure. Synthesis of the stereoisomers of semiochemicals has clarified their structure-bioactivity relationships to reveal the unprecedented diversity in the stereochemical aspects of pheromone communications. PMID- 10673319 TI - Femtosecond studies of electron dynamics at interfaces. AB - A delocalized electron at a metal-dielectric interface interacts with the adlayer and spatially localizes or self-traps on the femtosecond time scale into what is termed a small polaron. The dynamics can be studied by two-photon photoemission. Theoretical and experimental analyses reveal the interaction energy and the lattice vibrational mode that mediates electron localization. These results contribute to a fundamental understanding of electron behavior in weakly bonded solids and can lead to a better understanding of carrier dynamics in many different systems, including organic light-emitting diodes. PMID- 10673320 TI - What do titano- and zirconocenes do with diynes and polyynes? AB - A thorough investigation of the reactions of diynes, R(C&tbd1;C)(2)R, and polyynes, R(C&tbd1;C)(n)()R, with titanocene and zirconocene was performed using the metallocene sources Cp(2)M(L)(eta(2)-Me(3)SiC&tbd1;CSiMe(3)) (M = Ti, L = -; M = Zr, L = THF, pyridine). The conversions show an array of different products generated in complexation, coupling, and cleavage reactions. These results include remarkable structures (e.g., five-membered metallacyclocumulenes) and reactions (e.g., C-C single-bond cleavage). In addition, the first C-C single bond metathesis in homogeneous solution was discovered. The presented findings have been supported by theoretical studies. PMID- 10673321 TI - Nicotinic acetylcholine receptor agonists promote survival and reduce apoptosis of chick ciliary ganglion neurons. AB - The abundance, diversity, and ubiquitous expression of neuronal nicotinic acetylcholine receptors (AChRs) suggest that many are involved in functions other than synaptic transmission. We now report that a major AChR class promotes neuronal survival. The 10-day survival of ciliary ganglion neurons in basal culture medium (MEM) was approximately 35%, but increased to approximately 75% in MEM containing nicotine (MEM/Nic) or carbachol, an effect similar to that achieved by chronic depolarization with KCl. Pharmacological experiments revealed that agonist-enhanced survival requires activation of AChRs sensitive to alpha bungarotoxin (alphaBgt). alphaBgt-AChRs partly support neuronal survival by limiting apoptosis since fewer apoptotic neurons were observed in MEM/Nic compared to MEM. Moreover, nicotinic survival support was not further enhanced by fibroblast growth factor, as seen for KCl, but increased to 100% by adding PACAP, a trophic neuropeptide present in the ganglion. These results indicate that alphaBgt-AChR activation regulates neuronal survival and suggest a mechanism involving reduced apoptosis and interaction with an endogenous neuropeptide growth factor. PMID- 10673322 TI - Eph receptors and ephrin expression in cranial motor neurons and the branchial arches of the chick embryo. AB - Cranial motor axons navigate along a variety of pathways to their targets in the periphery of the head. Whereas somatic motor axons innervate tongue and eye muscles, visceral motor axons innervate parasympathetic ganglia, and branchiomotor axons innervate the branchial arches. The formation of these diverse pathways must depend upon molecules present in the environment traversed by growing axons. We have analyzed the potential roles of the ephrin ligands and their Eph tyrosine kinase receptors during cranial motor neuron development and axon pathfinding, by investigating expression patterns of these molecules at relevant stages in the chick. We detected expression of EphA3 and EphA4 among trigeminal and facial motor neurons, at times when these neurons are projecting to their muscle targets in the branchial arches. Corresponding ephrin-A ligands for these receptors were found to be expressed in specific regions of the arches during the same period, implicating ephrin-mediated interactions in cranial motor axon pathfinding. PMID- 10673323 TI - Sema3C and netrin-1 differentially affect axon growth in the hippocampal formation. AB - The interaction between outgrowing neurons and their targets is a central element in the development of the afferent and efferent connections of the hippocampal system. This requires that axonal growth cones recognize specific guidance cues in the appropriate target area. At present, little is known about the mechanisms that determine the lamina-specific termination of hippocampal afferents. In order to understand the role of different guidance factors, we analyzed the effects of Sema3C and Netrin-1 on explants from the entorhinal cortex, dentate gyrus, cornu ammonis regions CA1 and CA3 and medial septum in a collagen coculture assay. Our observations suggest that both semaphorins and netrin play important roles in the neuron-target interactions in the hippocampal system. Sema3C is involved in the control of the ingrowth of the septohippocampal projection. We also show that netrin-1 is involved in attracting commissural neurons from dentate gyrus/hilus and CA3 to their target area in the contralateral hippocampus. PMID- 10673324 TI - Inhibition of PKB/Akt1 by C2-ceramide involves activation of ceramide-activated protein phosphatase in PC12 cells. AB - Accumulation of ceramide has been reported in stress- and receptor-induced apoptosis in the nervous system. However, its role in apoptosis signaling remains elusive. We describe here the inhibition of the NGF-activated phosphoinositide 3 kinase (PI3K)-PKB/Akt1 survival pathway by the cell permeable analog C2-ceramide. C2-ceramide did not inhibit ERK, PI3K, or PDK1 activities and did not alter the translocation of PDK1 and Akt1 to the plasma membrane, but blocked nuclear translocation of Akt1. Down-regulation of the Akt pathway was due to enhanced dephosphorylation of Akt1 at residues T308 and S473. Moreover, Akt1 was dephosphorylated in vitro by a cation-independent phosphatase involving ceramide activated protein phosphatase (CAPP). Membrane-anchored Akt1 was more resistant to dephosphorylation/inactivation by C2-ceramide than wild-type Akt1. Consistently, N-myristylated-Akt1 conferred resistance to the apoptosis induced by C2-ceramide in PC12 cells. These results provide a novel mechanism for induction of apoptosis by ceramide in nerve-derived cells. PMID- 10673325 TI - Activating transcription factor 3 (ATF3) induction by axotomy in sensory and motoneurons: A novel neuronal marker of nerve injury. AB - Activating transcription factor 3 (ATF3), a member of ATF/CREB family of transcription factors, is induced in a variety of stressed tissue. ATF3 regulates transcription by binding to DNA sites as a homodimer or heterodimer with Jun proteins. The purpose of this study was to examine the expression and regulation of ATF3 after axonal injury in neurons in dorsal root ganglia (DRG) and spinal cord. In naive rats, ATF3 was not expressed in the DRG and spinal cord. Following the cut of peripheral nerve, ATF3 was immediately induced in virtually all DRG neurons and motoneurons that were axotomized, and the time course of induction was dependent on the distance between the injury site and the cell body. Double labeling using immunohistochemistry revealed that the population of DRG neurons expressing ATF3 included those expressing c-jun, and in motoneurons ATF3 and c jun were concurrently expressed after axotomy. In contrast to c-jun, ATF3 was not induced transsynaptically in spinal dorsal horn neurons. We conclude that ATF3 is specifically induced in sensory and motoneurons in the spinal cord following nerve injury and should be regarded as an unique neuronal marker of nerve injury in the nervous system. PMID- 10673326 TI - Agrin binds to beta-amyloid (Abeta), accelerates abeta fibril formation, and is localized to Abeta deposits in Alzheimer's disease brain. AB - Agrin is an extracellular matrix heparan sulfate proteoglycan (HSPG) well known for its role in modulation of the neuromuscular junction during development. Although agrin is one of the major HSPGs of the brain, its function there remains elusive. Here we provide evidence suggesting a possible function for agrin in Alzheimer's disease brain. Agrin protein binds the amyloidogenic peptide Abeta (1 40) in its fibrillar state via a mechanism that involves the heparan sulfate glycosaminoglycan chains of agrin. Furthermore, agrin is able to accelerate Abeta fibril formation and protect Abeta (1-40) from proteolysis, in vitro. Supporting a biological significance for these in vitro data, immunocytochemical studies demonstrate agrin's presence within senile plaques and cerebrovascular amyloid deposits, and agrin immunostained capillaries exhibit pathological alterations in AD brain. These data therefore suggest that agrin may be an important factor in the progression of Abeta peptide aggregation and/or its persistence in Alzheimer's disease brain. PMID- 10673327 TI - In vivo protection of nigral dopamine neurons by lentiviral gene transfer of the novel GDNF-family member neublastin/artemin. AB - The glial cell line-derived neurotrophic factor (GDNF)-family of neurotrophic factors consisted until recently of three members, GDNF, neurturin, and persephin. We describe here the cloning of a new GDNF-family member, neublastin (NBN), identical to artemin (ART), recently published (Baloh et al., 1998). Addition of NBN/ART to cultures of fetal mesencephalic dopamine (DA) neurons increased the number of surviving tyrosine hydroxylase (TH)-immunoreactive neurons by approximately 70%, similar to the maximal effect obtained with GDNF. To investigate the neuroprotective effects in vivo, lentiviral vectors carrying the cDNA for NBN/ART or GDNF were injected into the striatum and ventral midbrain. Three weeks after an intrastriatal 6-hydroxydopamine lesion only about 20% of the nigral DA neurons were left in the control group, while 80-90% of the DA neurons remained in the NBN/ART and GDNF treatment groups, and the striatal TH immunoreactive innervation was partly spared. We conclude that NBN/ART, similarly to GDNF, is a potent neuroprotective factor for the nigrostriatal DA neurons in vivo. PMID- 10673328 TI - Hemizygosity of delta-catenin (CTNND2) is associated with severe mental retardation in cri-du-chat syndrome. AB - Delta-catenin is an adherens junction protein involved in cell motility and expressed early in neuronal development. It was discovered as an interactor with presenilin-1. The genomic structure of the human delta-catenin gene (Human Gene Nomenclature Committee-approved symbol CTNND2) was determined and mapped to 5p15.2. A deletion of this chromosomal region has been associated with the cri-du chat syndrome (CDCS), a segmental aneusomy syndrome of 5p that is associated with an unusual high-pitched cry at birth, facial dysmorphology, poor growth, and severe mental retardation. delta-catenin maps to a specific region in 5p15.2 that has been implicated in the mental retardation phenotype. The breakpoints in patients with 5p terminal deletions were characterized with respect to the severity of mental retardation and the physical location of the delta-catenin gene. A strong correlation was found between the hemizygous loss of delta-catenin and severe mental retardation. These findings and the properties of delta-catenin as a neuronal-specific protein, expressed early in development and involved in cell motility, support its role in the mental retardation of CDCS when present in only one copy. PMID- 10673329 TI - Genomic and expression analyses of alternatively spliced transcripts of the MLL septin-like fusion gene (MSF) that map to a 17q25 region of loss in breast and ovarian tumors. AB - We previously defined a common region of 17q25 loss in breast and ovarian tumors, suggesting localization of at least one putative tumor suppressor gene. Genomic clones from the interval were used to isolate candidate transcripts. One novel transcript had strong homology to a septin family of GTPase genes involved in cytokinesis. This gene was recently identified as a myeloid/lymphoid leukemia (MLL) fusion protein partner in acute myeloid leukemia and was named MSF (MLL septin-like fusion). As this gene may play roles in both leukemogenesis and tumorigenesis, it is essential to understand its structure and normal expression. We cloned two human alternative transcripts and identified a third database variant of MSF. RNA expression studies with a probe common to the three novel sequences showed differential expression of 4.0- and 3.0-kb transcripts in all adult and fetal tissues tested. A probe spanning sequence unique to one MSF variant detected specific expression of the 4.0-kb transcript in all tissues. Another probe unique to a different MSF variant detected a 4.0-kb transcript only in skeletal muscle. Proteins of 422 and 586 amino acids were predicted from the novel alternate transcripts and included both a xylose isomerase 1 domain and a GTPase domain. Nine common exons, three alternatively spliced exons, and six polymorphisms were identified. PMID- 10673330 TI - The alpha-fetoprotein promoter is the target of Afr1-mediated postnatal repression. AB - The alpha-fetoprotein (AFP) gene is transcribed at high levels in the fetal liver and is repressed at birth, leading to low but detectable levels of AFP mRNA in the adult liver. This repression is regulated, in part, by a locus that is unlinked to AFP called Alpha-fetoprotein regulator 1 (Afr1). Previous studies showed that Afr1 regulation is independent of the AFP enhancers but requires the 1-kb AFP promoter/repressor region. Here, we demonstrate that a transgene with the 250-bp AFP promoter region linked to AFP enhancer element EII is expressed in the fetal liver and is postnatally repressed. In addition, this transgene is regulated by Afr1. These data indicate that the promoter is involved in postnatal AFP repression. Furthermore, we provide a high-resolution map of the Afr1 locus on mouse chromosome 15. PMID- 10673331 TI - PEX7 gene structure, alternative transcripts, and evidence for a founder haplotype for the frequent RCDP allele, L292ter. AB - We recently reported cloning a cDNA encoding Pex7p, the peroxisomal PTS2 receptor. PEX7 mutations cause the peroxisome biogenesis disorder (PBD) rhizomelic chondrodysplasia punctata (RCDP). In a survey of 44 RCDP probands, we found that one PEX7 allele, L292ter, accounted for 50% of mutant PEX7 genes. Here we report the characterization of the PEX7 structural gene, which spans 102 kb on chromosome 6q21-q22.2 and contains at least 10 exons. In addition to the predominant full-length transcript, we identified eight smaller PEX7 transcripts generated by alternative exon splicing in several tissues. However, none of these splice forms was able to restore PTS2 protein import into peroxisomes when expressed in RCDP fibroblasts nor did they inhibit PTS2 protein import when expressed in normal fibroblasts. To determine whether the high frequency of the L292ter allele is due to a founder effect, we identified five polymorphic markers (four diallelic markers and one CA repeat) spanning the PEX7 gene. We show that all 12 L292ter homozygotes in our patient sample have an identical haplotype at these five sites, consistent with the hypothesis that the L292ter mutation arose once on an ancestral chromosome in the Caucasian population. PMID- 10673332 TI - Identification of a murine locus conveying susceptibility to cadmium-induced forelimb malformations. AB - The heavy metal cadmium (Cd), an environmentally ubiquitous contaminant, is a potent teratogen in mice. When administered parenterally, it induces an array of malformations that vary in scope and severity with the route, dose, time of administration, and the strain of the animal. When administered intraperitoneally on day 9.0 of gestation, 4 mg/kg cadmium chloride produces forelimb defects (predominantly ectrodactyly) in over 80% of fetuses of the C57BL/6 mouse strain, while no limb defects are observed in the identically treated SWV strain. Like other examples of strain-specific teratogenic activity, the underlying nature of the differential susceptibility remains unknown. The present study investigates the segregation of sensitivity to Cd-induced forelimb defects in crosses between C57BL/6 and SWV mice and provides evidence for the involvement of both maternal and fetal factors in the determination of defect expression. In addition, quantitative trait loci (QTL) analysis of the fetal genetic component was performed among 198 backcross progeny, utilizing a genomic linkage map of 149 informative microsatellite markers. One QTL demonstrating significant linkage to expression of the defect, designated Cadfar (cadmium-induced forelimb autopod reduction), was mapped to the distal end of chromosome 6 with a lod score of 3.1. PMID- 10673333 TI - An integrated genetic linkage map with 1,137 markers constructed from five F2 crosses of autoimmune disease-prone and -resistant inbred rat strains. AB - The rat (Rattus norvegicus) is an important experimental model for many human diseases including arthritis, diabetes, and other autoimmune and chronic inflammatory diseases. The rat genetic linkage map, however, is less well developed than those of mouse and human. Integrated rat genetic linkage maps have been previously reported by Pravenec et al. (1996, Mamm. Genome 7: 117-127) (500 markers mapped in one cross), Bihoreau et al. (1997, Genome Res. 7: 434-440) (767 markers mapped in three crosses), Wei et al. (1998, Mamm. Genome 9: 1002-1007) (562 markers mapped in two crosses), Brown et al. (1998, Mamm. Genome 9: 521-530) (678 markers mapped in four crosses), and Nordquist et al. (1999, Rat Genome 5: 15-20) (330 markers mapped in two crosses). The densest linkage map combined with a radiation hybrid map, reported by Steen et al. (1999, Genome Res. 9: AP1-AP8), includes 4736 markers mapped in two crosses. Here, we present an integrated linkage map with 1137 markers. We have constructed this map by genotyping F2 progeny of five crosses: F344/NHsd x LEW/NHsd (673 markers), DA/Bkl x F344/NHsd (531 markers), BN/SsN x LEW/N (714 markers), DA/Bkl x BN/SsNHsd (194 markers), and DA/Bkl x ACI/SegHsd (245 markers). These inbred rat strains vary in susceptibility/resistance to multiple autoimmune diseases and are used extensively for many types of investigation. The integrated map includes 360 loci mapped in three or more crosses. The map contains 196 new SSLP markers developed by our group, as well as many SSLP markers developed by other groups. Two hundred forty genes are incorporated in the map. This integrated map should allow comparison of rat genetic maps from different groups and thereby facilitate genetic studies of rat autoimmune and related disease models. PMID- 10673334 TI - Sequence, structure, and evolution of a complete human olfactory receptor gene cluster. AB - The olfactory receptor (OR) gene cluster on human chromosome 17p13.3 was subjected to mixed shotgun automated DNA sequencing. The resulting 412 kb of genomic sequence include 17 OR coding regions, 6 of which are pseudogenes. Six of the coding regions were discovered only upon genomic sequencing, while the others were previously reported as partial sequences. A comparison of DNA sequences in the vicinity of the OR coding regions revealed a common gene structure with an intronless coding region and at least one upstream noncoding exon. Potential gene control regions including specific pyrimidine:purine tracts and Olf-1 sites have been identified. One of the pseudogenes apparently has evolved into a CpG island. Four extensive CpG islands can be discerned within the cluster, not coupled to specific OR genes. The cluster is flanked at its telomeric end by an unidentified open reading frame (C17orf2) with no significant similarity to any known protein. A high proportion of the cluster sequence (about 60%) belongs to various families of interspersed repetitive elements, with a clear predominance of LINE repeats. The OR genes in the cluster belong to two families and seven subfamilies, which show a relatively high degree of intermixing along the cluster, in seemingly random orientations. This genomic organization may be best accounted for by a complex series of evolutionary events. PMID- 10673335 TI - Molecular cloning of a novel apoptosis-related gene, human Nap1 (NCKAP1), and its possible relation to Alzheimer disease. AB - Expression profiles of thousands of genes (cDNAs) were analyzed in sporadic Alzheimer disease (AD)-affected brains in comparison with normal subjects by using the high-density cDNA filter method and differential display analysis. Among 31 differentially expressed genes, one gene was found to be markedly depressed in AD-affected brains. A full-length (or nearly full-length) cDNA of the gene was isolated and sequenced. The cDNA turned out to be an orthologue of rat Nap1. The gene was thus designated human Nap1 (HGMW-approved symbol NCKAP1) and was mapped to human chromosome 2q32 by fluorescence in situ hybridization. Northern blotting and in situ hybridization studies showed that in brain, the gene is predominantly expressed in neuronal cells. Antisense oligo DNA of human Nap1 transcripts was found to induce apoptosis of neuronal cells. Based on these results, the possible role of human Nap1 in AD is discussed. PMID- 10673336 TI - EHD2, EHD3, and EHD4 encode novel members of a highly conserved family of EH domain-containing proteins. AB - Exon trapping from a bacterial artificial chromosome (BAC 78138) mapping to the 19q13.3 glioma tumor suppressor candidate region yielded two exons that recognized a 3.6-kb transcript on Northern blot. Screening of a human fetal brain cDNA library with these exons identified three novel genes, designated EHD2, EHD3, and EHD4, which are homologous to the recently characterized human EHD1 (testilin/HPAST) and its mouse homolog Ehd1, as well as to homologs in Drosophila (Past1) and Caenorhabditis elegans. Alignment of the predicted peptide sequences revealed striking similarities, with multiple conserved regions that include a nucleotide-binding consensus site at the N-terminus, a bipartite nuclear localization signal, and an eps15 homology (EH) protein-binding domain with an EF hand motif at the C-terminus. The genes are specifically expressed, with EHD2 highly expressed in heart, EHD3 in brain and heart, and EHD4 in heart and pancreas. EHD2 was confirmed to originate from BAC 78138 at 19q13.3; radiation hybrid mapping localized EHD3 and EHD4 to 2p21 and 15q11.1, respectively; EHD1 has been previously mapped to 11q13. The three EHD1 paralogs therefore represent novel members of a family of human EH domain-containing proteins that may play a role in endocytosis and signaling. Mutation analysis of the five coding exons of EHD2 in gliomas failed to detect any tumor-specific alterations, thus indicating that EHD2 is an unlikely candidate for the 19q tumor suppressor gene. PMID- 10673337 TI - Structure, expression, and chromosome mapping of LATS2, a mammalian homologue of the Drosophila tumor suppressor gene lats/warts. AB - We have cloned and characterized LATS2, a novel mammalian homologue of the Drosophila tumor suppressor gene lats/warts. Northern blot analysis showed ubiquitous expression of mouse LATS2 (MmLATS2) mRNA, whereas expression of human LATS2 (HsLATS2) mRNA was enhanced in skeletal muscle and heart. Immunoblotting analysis of fractionated cell lysates showed HsLats2 to be a nuclear protein. We mapped the MmLATS2 gene to mouse chromosome 14 by interspecific backcross analysis. We also mapped the HsLATS2 gene (by fluorescence in situ hybridization) to the 13q11-q12 region, in which a loss of heterozygosity has been frequently observed in many primary cancers and to which the tumor suppressor genes RB and BRCA2 have also been mapped. PMID- 10673338 TI - Genetic refinement and physical mapping of the CMT4B gene on chromosome 11q22. AB - Charcot-Marie-Tooth disease type 4B (CMT4B) is a demyelinating autosomal recessive motor and sensory neuropathy characterized by focally folded myelin sheaths in the peripheral nerve. We recently mapped the CMT4B gene to a 5-cM interval on chromosome 11q22, using homozygosity mapping and haplotype sharing analysis on a large inbred pedigree. In the present study, we report the construction of a YAC-based transcript map across the 5-cM critical region, including 26 YACs, 35 STSs, and 52 ESTs. Furthermore, using 15 additional physically ordered microsatellite markers from the 11q22 region on the original inbred family, we were able to narrow the critical interval for the gene to 2 Mb, which is now flanked by markers D11S1757 and CHLC-GATA3B05. Finally, after computer analysis of the 33 ESTs assigned to the 2-Mb interval, we demonstrated that 21 different transcripts as well as 3 known genes might represent potential candidates for the disease. PMID- 10673339 TI - Cloning, characterization, and chromosomal localization of Pnck, a Ca(2+)/calmodulin-dependent protein kinase. AB - Calcium is an important second messenger in eukaryotic cells. Many of the effects of calcium are mediated via its interaction with calmodulin and the subsequent activation of Ca(2+)/calmodulin-dependent (CaM) kinases. CaM kinases are involved in a wide variety of cellular processes including muscle contraction, neurotransmitter release, cell cycle control, and transcriptional regulation. While CaMKII has been implicated in learning and memory, the biological role of the other multifunctional CaM kinases, CaMKI and CaMKIV, is largely unknown. In the course of a degenerate RT-PCR protein kinase screen, we identified a novel serine/threonine kinase, Pnck. In this report, we describe the cloning, chromosomal localization, and expression of Pnck, which encodes a 38-kDa protein kinase whose catalytic domain shares 45-70% identity with members of the CaM kinase family. The gene for Pnck localizes to mouse chromosome X, in a region of conserved synteny with human chromosome Xq28 that is associated with multiple distinct mental retardation syndromes. Pnck is upregulated during intermediate and late stages of murine fetal development with highest levels of expression in developing brain, bone, and gut. Pnck is also expressed in a tissue-specific manner in adult mice with highest levels of expression detected in brain, uterus, ovary, and testis. Interestingly, Pnck expression in these tissues is restricted to particular compartments and appears to be further restricted to subsets of cells within those compartments. The chromosomal localization of Pnck, along with its tissue-specific and restricted pattern of spatial expression during development, suggests that Pnck may be involved in a variety of developmental processes including development of the central nervous system. PMID- 10673340 TI - Isolation and characterization of a novel human paired-like homeodomain containing transcription factor gene, VSX1, expressed in ocular tissues. AB - Homeodomain transcription factors control cell fates during the development of all animals. The paired-like subfamily of homeodomain proteins has been particularly implicated in ocular development in different species. In this paper we report the cDNA sequence, genomic structure, localization, and expression data of a novel paired-like homeobox-containing gene, VSX1, isolated from a human embryonic craniofacial cDNA library using the degenerate-PCR approach. The composed VSX1 cDNA sequence of 1433 bp was predicted to encode a protein of 365 amino acid residues. Maximal homology at the protein level was identified with the paired-like homeoproteins of the CVC-domain family: 92-97% identity was seen in the homeodomain region with 55% overall identity to zebrafish and goldfish Vsx1 and 35% overall identity to goldfish Vsx2 and murine Chx10. The gene was found to consist of five exons that are distributed over 6.2 kb of genomic sequence. VSX1 was localized to the 20p11-q11 region, which is homologous with the distal part of mouse chromosome 2. Expression of VSX1 was detected in embryonic craniofacial and adult ocular tissues. Several ocular phenotypes have been mapped to the VSX1 region in both human and mouse genomes, and its candidacy for these disorders is discussed. PMID- 10673341 TI - Structure, expression, and chromosomal localization of the mouse Atox1 gene. AB - Copper trafficking in eukaryotes involves small proteins termed metallochaperones, which mediate copper delivery to specific intracellular sites. Previous studies in yeast and human cell lines have suggested that Atox1 plays a critical role in copper delivery to the secretory pathway. In the present study, a mouse Atox1 (mAtox1) cDNA was cloned and shown to encode an open reading frame with 85% amino acid identity to human Atox1. RNA blot analysis revealed that mAtox1 was expressed as a single transcript in multiple tissues, and immunoblotting indicated that the relative abundance of mAtox1 mRNA directly correlated with mAtox1 protein. Analysis of the mAtox1 gene locus revealed a genomic structure with four exons encompassing a total of 14.5 kb. RFLP and haplotype analyses indicated that the mAtox1 locus was tightly linked to the Trhr and D15Bir7 loci on mouse chromosome 15. Taken together, these data reveal marked evolutionary conservation of Atox1 structure and provide a genomic organization and localization that will aid in the genetic deciphering of the molecular role of this protein in copper homeostasis. PMID- 10673342 TI - Litomosoides sigmodontis: dynamics of the survival of microfilariae in resistant and susceptible strains of mice. AB - Litomosoides sigmodontis in the BALB/c mouse is the only model of filariasis which allows the observation of the complete development in an immunocompetent mouse. In this study, we injected microfilariae (mf) intravenously, as well as into the pleural cavity, the site of natural release of mf from adult female worms, and followed the kinetics of elimination within the host. In susceptible BALB/c mice, mf circulated at high levels in the blood. In contrast, in C57BL/6 mice, which are refractory to full development, mf were eliminated rapidly from the peripheral blood. However, 6 days after intrapleural injection, viable larvae could be found in the pleural cavity and lung capillaries of both susceptible and resistant strains. The numbers of mf in the pleural cavity and lung capillaries in individual mice were significantly correlated, but not dependent on strain or peripheral microfilaraemia. Thus, although C57BL/6 mice showed enhanced production of nitric oxide by pleural exudate cells and a faster change in the numbers of circulating leukocytes after injection, rapid killing of mf by cell or nitric oxide-mediated mechanisms were not the reason for the different outcome. Furthermore, 3 h after iv injection, only a small percentage of mf could be recovered from the peripheral circulation, indicating the presence of a reservoir for mf containment. In conclusion, injected mf showed disparate dynamics of persistence within susceptible and resistant hosts, which is similar to the disparate outcome of natural infections with L. sigmodontis. This difference became obvious within 1 day after injection. The lung capillary system plays obviously a crucial part in regulation of microfilaremia. Our model also provides a possible means to explain frequent cases of occult infections in human filariasis. PMID- 10673343 TI - Schistosoma mansoni: differential expression of cathepsins L1 and L2 suggests discrete biological functions for each enzyme. AB - Schistosoma mansoni cathepsins L1 (SmCL1) and L2 (SmCL2) were expressed as active recombinant proteinases in Saccharomyces cerevisiae. The recombinant enzymes exhibited substrate preferences characteristic of cathepsin-L-like cysteine proteinases. However, the enzymes differed in their substrate specificities; SmCL1 cleaved Boc-Val-Leu-Lys-NHMec with a higher efficiency than it cleaved Z Phe-Arg-NHMec, whereas the opposite was true for SmCL2. The enzymes also differed in their pH profiles of activity; SmCL1 exhibited a broad pH profile with an optimum of pH 6. 5, while SmCL2 was active only in the acidic pH range with an optimum of 5.35. Immunoblot and RT-PCR analyses revealed that the native forms of both SmCL1 and SmCL2 are expressed in male and female worms, but at higher levels in adult female compared to male schistosomes. Additionally, both enzymes were observed in the excretory/secretory products of adult worms. The RT-PCR analysis indicated that neither enzyme is expressed in S. mansoni eggs or in miracidia, suggesting that the cathepsin-L-like activity that has been previously reported to be expressed in these stages may be the product of another gene(s). Cercariae do not express SmCL2, but appear to express SmCL1 in its inactive precursor form. Together with the findings of previous immunolocalization and phylogenetic analyses, the results reported here demonstrate that SmCL1 and SmCL2 are distinct cathepsin cysteine proteinases and strongly suggest that they play discrete biological roles. PMID- 10673344 TI - Trypanosoma cruzi: the effect of nitric oxide synthesis inhibition on the CD4 T cell response to the trans-sialidase superfamily. AB - During Trypanosoma cruzi infection the trans-sialidase superfamily stimulates the development of a large population of CD4 T lymphocytes that produces IFNgamma. These CD4 T cells fail to proliferate when stimulated in vitro. Why they fail to proliferate remains unclear. Nitric oxide is a critical component of the host immune response against T. cruzi, and to determine if NO inhibits trans-sialidase superfamily-specific proliferative responses, mice were fed either N(G)-nitro-L arginine methylester (L-NAME), an inhibitor of inducible nitric oxide synthase (iNOS), or N(G)-nitro-D-arginine methyl ester (D-NAME), an inactive analog of L NAME. The L-NAME-fed mice had increased parasitemia and mortality compared to the D-NAME-fed mice. Following stimulation with a T. cruzi trans-sialidase superfamily protein, splenocytes from both groups of mice failed to proliferate but continued to make similar amounts of IFNgamma, suggesting that the development of the trans-sialidase superfamily-specific CD4 response was not affected by iNOS inhibition. In addition, IL-2 receptor (IL-2R) expression was increased on T cells isolated from L-NAME-fed mice. These data suggest that during T. cruzi infection NO causes downregulation of IL-2R expression, but does not cause inhibition of trans-sialidase superfamily-specific CD4 T cell proliferation. Rather, the trans-sialidase superfamily proliferation may be inhibited by epitope variation. PMID- 10673345 TI - Brugia malayi: localization of nitric oxide synthase in a lymphatic filariid. AB - Nitric oxide synthase converts L-arginine to citrulline and nitric oxide, a gaseous signaling molecule critical to multiple physiological responses. Nitric oxide synthase was detected by Western blot analysis of Brugia malayi extracts using an antibody raised against a peptide from murine brain nitric oxide synthase. Using NADPH diaphorase staining and immunohistochemistry, nitric oxide synthase was localized in the parasitic nematode B. malayi. As in Ascaris suum, nitric oxide synthase was detected in the body wall muscles of adult B. malayi. This localization pattern is in agreement with the role of nitric oxide in the control of muscle tone in other invertebrates and in vertebrates. A novel finding was the localization of nitric oxide synthase in the oocytes, in developing embryos, and in spermatozoa. B. malayi nitric oxide synthase may play a role in developmental signaling, as has been suggested for Drosophila and Ilyanassa, a marine mud snail. PMID- 10673346 TI - Fasciola hepatica: parasite-secreted proteinases degrade all human IgG subclasses: determination of the specific cleavage sites and identification of the immunoglobulin fragments produced. AB - The study was focused on the relationship of Fasciola hepatica-secreted proteinases and human IgG subclasses. Each IgG was incubated at different pH values and lengths of time with either the adult parasite excretion-secretion products or the purified cysteinyl proteinases cathepsin L1 and cathepsin L2. The Ig fragments produced were isolated and characterized by Western blot analysis, and the specific cleavage sites were determined by amino acid sequence analysis. Parasite excretion-secretion products and both cathepsins L produced similar degradation patterns and cleaved all human IgG subclasses at the hinge region, yielding at pH 7.3 and 37 degrees C Fab and Fc fragments in the case of IgG1 and IgG3 or Fab(2) and Fc in IgG2 and IgG4. While IgG1 and IgG3 were readily degraded by E/S products either in the presence or in the absence of reducing agents, IgG2 and IgG4 were resistant to proteolysis and were only digested in the presence of 0.1 M dithiothreitol. The cathepsins L needed the presence of dithiothreitol to digest IgG1, IgG2, and IgG4 whereas IgG3 was identically cleaved under both reducing and nonreducing conditions. The main cleavage sites produced by E/S products, CL1, or CL2 were located at the positions peptide bonds: His237-Thr238, Glu237-Cys239, Gly233-Asp234, and Ser241-Cys242 for gamma1, gamma2, gamma3, or gamma4, respectively. The enzymes gave additional splitting sites on the middle hinge of IgG3 to produce shorter Fc fragments and also produce Fd degradation of the IgG4. No cleavage specificity differences were found between CL1 and CL2, but they differed in the kinetics of IgG3 degradation. By lowering the pH, only the E/S products produced concomitant destruction of the Fc while preserving the Fab portion. Under all the conditions assayed the enzymes produced an Fc'-like fragment of 14-15 kDa corresponding to the whole CH3 domain of the immunoglobulin. Contrary to the extensive degradation produced by cathepsins on digested proteins, its actions on IgG subclasses were specific and restricted; thus, all the fragments produced could be potentially involved in the mechanisms used by the parasite to evade the host immune response. PMID- 10673347 TI - Nitric oxide synthase and cGMP activity in the salivary glands of the American dog tick Dermacentor variabilis. AB - We colocalized nitric oxide synthase (NOS) activity in epithelial cells that surround the salivary gland duct in female Dermacentor variabilis with NADPH diaphorase histochemistry and immunohistochemistry using a polyclonal anti endothelial NOS. Using size-exclusion chromatography, a fraction with a molecular mass of about 185 kDa that had diaphorase activity was eluted from tick salivary gland homogenate. This fraction converted arginine to citrulline with the production of nitric oxide (NO), which was detected by using electron spin resonance spectroscopy. The complete activity of the diaphorase fraction was dependent on NADPH, FAD, tetrahydrobiopterin, calmodulin, (CaM), and Ca(2+), but was not dependent on dithiothreitol. The arginine analog N(G)-monomethyl-L arginine inhibited the activity of this fraction. NO and arginine activated soluble guanylate cyclase to produce cGMP in dopamine-stimulated isolated salivary glands. Dopamine-stimulated isolated salivary glands treated with tick saline containing either EDTA, the NOS inhibitor N(G)-nitro-L-arginine methyl ester, or the calcium/CaM binding inhibitor W-7 showed no increase in cGMP. The NO donor sodium nitroprusside significantly increased cGMP levels in unstimulated isolated salivary glands. A possible function for NO in salivation by this ixodid tick is discussed. PMID- 10673348 TI - Plasmodium falciparum: effective use of the CO(2)-NaHCO(3) buffer system for evaluating chloroquine resistance. PMID- 10673349 TI - Trichomonas vaginalis: detection of nucleoside hydrolase activity as a potential screening procedure. PMID- 10673350 TI - Trypanosoma cruzi: molecular cloning of a gene coding for a putative vacuolar protein. AB - We describe the characterization of Tc38, a Trypanosoma cruzi gene coding for a 337-amino-acid protein with a predicted molecular mass of 38 kDa. Tc38 presents similarities to the plant storage vacuolar protein gamma-3-hordein involved in the transport and targeting of prolamins to the vacuole of developing barley endosperm. Western blot analysis using a polyclonal antiserum against recombinant Tc38 revealed that the protein is differentially expressed in the different life stages of the parasite, showing a higher expression in the epimastigote and tripomastigote stages. Immunofluorescence studies suggest that the protein is located in putative vacuolar structures in epimastigotes. The functionality of this protein in T. cruzi remains to be elucidated. PMID- 10673351 TI - Could Nef and Vpr proteins contribute to disease progression by promoting depletion of bystander cells and prolonged survival of HIV-infected cells? AB - A growing body of literature suggests that the HIV accessory proteins Nef and Vpr could be involved in depletion of CD4(+) and non-CD4(+) cells and tissue atrophy, and in delaying the death of HIV-infected cells. Cell depletion is likely to be predominantly a bystander effect because the number of cells dying far outnumbers HIV-infected cells and is not confined to CD4(+) cells. The myristylated N terminal region of Nef has severe membrane disordering properties, and when present in the extracellular medium causes rapid lysis in vitro of a wide range of CD4(+) and non-CD4(+) cells, suggesting a role for extracellular Nef in the depletion of bystander cells. A direct role for HIV-1 Nef in cytopathicity is supported by studies in HIV-infected Hu Liv/Thy SCID mice, in transgenic mice expressing nef gene alone, and in rhesus macaques infected with SIV/HIV chimeric virus containing HIV-1 nef. The N-terminal region of Nef has been directly implicated in development of simian AIDS. Extracellular Vpr and C-terminal fragments of Vpr cause membrane permeabilization and apoptosis of a wide range of CD4(+) and non-CD4(+) cells, and could also contribute to depletion of bystander cells. A direct in vivo role for Vpr in thymocyte depletion, thymic atrophy, and nephropathy is suggested in studies with vpr transgenic mice. Intracellular Nef and Vpr could help HIV-infected cells evade cell death by inhibiting apoptosis of infected cells and by avoiding virus-specific CTL response. Nef and Vpr are potential targets for therapeutic intervention and vaccine development, and strategies that prevent the death of bystander cells while promoting the early death of HIV-infected cells could arrest or retard progression to AIDS. PMID- 10673352 TI - Two peptides from CD23, including the inverse RGD sequence and its related peptide, interact with the MHC class II molecule. AB - The human CD23 molecule (low affinity receptor for IgE) has a C-type lectin domain, a reversed Arg-Gly-Asp (RGD) sequence near the C-terminus, and an "RGD binding inhibitory peptide" at the root of the N-sugar chain. Three peptides were synthesized to determine their functions, i.e., #1, including an inverse RGD sequence near the C-terminus; #2, RGD-binding inhibitory peptides in the gpIIIa chain of platelet integrin gpIIb/IIIa; and #3, the inverse sequence located at the root of the N-sugar chain of CD23 which has homology to peptide 2. Among the three peptide, only peptide 3 inhibited aggregation of L-KT9 cells. Isotope labeled peptides 1 and 3 bound to MHC class II molecules but peptide 1 did not bind to CD23 molecules. Peptide 3 showed a higher affinity to MHC class II than did peptide 1. Both peptides in CD23, therefore, seem to have interesting and important functions in relation to MHC class II molecules and also to CD23 molecules when CD23 on EBV-transformed B cells acts as a lectin in homotypic cell aggregation. The physiological function of CD23 was discussed from an evolutional point of view. PMID- 10673353 TI - Interferon-alpha signaling promotes nucleus-to-cytoplasmic redistribution of p95Vav, and formation of a multisubunit complex involving Vav, Ku80, and Tyk2. AB - Interferons (IFNs) are a family of hormone-like secretory proteins with multiple phenotypical changes, including gene expression and morphological alterations. Earlier studies have shown that IFN-activated Tyk2 kinase physical associates with p95Vav (Vav), a proto-oncogene gene product expressed in hematopoietic cells. Since Tyk2 is a cytoplasmic kinase and Vav is believed to be localized in the nuclear compartment, here we explored the possibility of Vav redistribution in IFN-alpha-activated cells, using the U266 human myeloma cell line as a model system. Using biochemical assays and in situ confocal microscopy, we demonstrate that IFN-alpha treatment triggers a rapid (10 min) translocation of Vav from the nuclear compartment to the cytoplasm. In addition, we also show the existence of IFN-alpha-induced physical interaction between Vav and Ku80, Ku80, and Tyk2, and among Vav, Ku80, and Tyk2 in the cytoplasmic compartment of IFN-stimulated cells. The observed IFN-alpha-induced association among Vav, Ku80, and Tyk2 was dependent on cellular tyrosine kinase activity. Since recently Vav has been shown to promote the GDP/GTP exchange activity of the cytoskeleton signaling molecule small GTPase Rac1 and activates its downstream signaling, our present findings raise the possibility of involvement of the small GTPase in IFN signaling leading to its biological effects, including cytoskeleton reorganization. PMID- 10673354 TI - CYRL, a novel cytokine receptor-like protein expressed in testis, lung, and spleen. AB - The interleukin-3 receptor is composed of a ligand-specific alpha subunit (IL 3Ralpha) and a beta subunit (beta(c) or beta(IL3)). Here we report the cloning of a rat brain cDNA transcript with significant homology to IL-3Ralpha, which we have termed CYRL, for CYtokine Receptor-Like protein. A number of conserved motifs identify CYRL as a member of the alpha family of cytokine receptor subunits, but the extracellular domain was too divergent from the mouse IL 3Ralpha sequence to suggest that CYRL is the rat ortholog of IL-3Ralpha. CYRL mRNA expression by Northern blotting was highest in the testis, intermediate in the lung, and modest in spleen, brain, and heart. Antibodies generated against the extracellular domain of CYRL specifically labeled a broad immunoreactive band of M(r) approximately 50,000 in membrane fractions of testis, lung, and spleen. CYRL appears to be a novel cytokine receptor alpha-subunit of unknown function and with no defined ligands. PMID- 10673355 TI - Identification of the promoter region of human placental 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase gene. AB - The placenta-type isozyme of human 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase (HP2K) is expressed in several tissues such as placenta, brain, testis, liver, kidney, skeletal muscle, and primary blood mononuclear cells. To better understand the regulation of HP2K gene expression, we isolated and characterized its genomic DNA, which includes the promoter region. The results of oligo-capping analysis indicate that the transcription start point (tsp) is an adenine residue 329 bp upstream of the translational start codon. DNA sequence analysis of this gene shows that the promoter region that contains the TATA box sequence and the 5'-UTR is different from the other known PFK-2/F2, 6BPase genes. In addition, its 5'-flanking and 5'-UTR both have G + C-rich sequences containing Sp1 binding sites. To identify the promoter/enhancer region of HP2K gene, we performed transfection analyses of human choriocarcinoma BeWo cells with HP2K promoter-luciferase constructs. These experiments identified a promoter region 164 bp upstream from the tsp and an enhancer region between -1265 and -1329 on the 5'-flanking sequences. We also showed that Sp1 sites were not essential for HP2K transcription. Following transfection, stimulation experiments with serum, progesterone and phorbol 12-myristate 13-acetate showed that only the construct with the enhancer containing putative early growth response-1 binding motif was responsive to serum. We propose that the transcription of HP2K is strictly controlled by tissue-specific factors even though its genomic DNA contains several transcriptional elements. PMID- 10673356 TI - Direct interaction between emerin and lamin A. AB - Emerin is the protein of the inner nuclear membrane that is affected by mutation in X-linked Emery-Dreifuss muscular dystrophy. The autosomal dominant form of the disease is caused by mutations in the lamin A/C gene. Several lines of circumstantial evidence have suggested an interaction of emerin with lamins in the nuclear lamina but direct interaction between the two proteins has not yet been demonstrated. We now demonstrate direct interaction between recombinant emerin and lamin A molecules using biomolecular interaction analysis (BIA) and monoclonal antibodies. An emerin-lamin A interaction system may be related in function to the LAP2-lamin B system at the inner nuclear rim. PMID- 10673357 TI - A conditioning lesion promotes in vivo nerve regeneration in the contralateral sciatic nerve of rats. AB - A conditioning lesion in the sciatic nerve increases in vivo axonal regeneration in the nerve after a second transection. We studied whether this increased regeneration also occurs in the contralateral nerve. The left sciatic nerve was transected and sutured in Wistar rats; the nerve was exposed but not transected in controls. After 5 days, the right sciatic nerves of all rats were transected and sutured. Neuronal regeneration was measured at 0, 1, 3, 5, and 7 days with the pinch test and histological staining. IL-1beta and TGF-beta1 expression was also measured. The initial delay in the experimental group was significantly shorter, but the regeneration rates were the same. The expression of IL-1beta and TGF-beta1 in the right dorsal root ganglia was significantly higher in the experimental group. Nerve injury enhances cytokine expression in the contralateral dorsal root ganglion and promotes contralateral nerve regeneration in vivo by shortening the initial delay. PMID- 10673358 TI - Internalization and processing of human angiogenin by cultured aortic smooth muscle cells. AB - Human angiogenin is a 14-kDa plasma protein with angiogenic and ribonucleolytic activities. Angiogenin binds specifically to aortic smooth muscle cells, activates second messenger pathways, and inhibits their proliferation. Human and bovine aortic smooth muscle cells were used to study the internalization and intracellular fate of human angiogenin at 37 degrees C. Using a specific antibody against angiogenin, we found that the internalized native protein was localized in the perinuclear region at 30 min and then dispersed throughout the cytoplasm. In conditions favoring receptor-mediated endocytosis, internalization of iodinated angiogenin showed a first peak at 5 min and then further increased for up to 24 h. The half-life of the molecule, calculated as 12 h in chase experiments, could contribute to its intracellular accumulation. In cell extracts, in addition to the 14-kDa protein, a 8.7-kDa fragment was observed at 24 h, and three fragments with molecular mass of 10.5, 8.7, and 6. 1 kDa were detected at 48 h. Our data point to a specific internalization and processing of human angiogenin by aortic smooth muscle cells. PMID- 10673359 TI - Tumor antigen HuR binds specifically to one of five protein-binding segments in the 3'-untranslated region of the neurofibromin messenger RNA. AB - 3'-untranslated regions of various mRNAs have been shown to contain sequence motifs which control mRNA stability, translatability, and efficiency of translation as well as intracellular localization. We aimed to identify protein binding regions of the long and highly conserved 3'UTR of the mRNA coding for neurofibromin, a well-known tumor suppressor protein, whose genetic deficiency causes the autosomal dominant disease neurofibromatosis type 1 (NF1). We discovered five RNA fragments that were able to undergo specific binding to proteins from cell lysates (NF1-PBRs, NF1-protein-binding regions). Additionally we identified the Elav-like protein HuR binding to NF1-PBR1. HuR interacts with AU-rich elements in the 3'UTR of many protooncogenes, cytokines, and transcription factors, thereby regulating the expression of these mRNAs on the posttranscriptional level. Transfection assays with a CAT reporter construct revealed reduced expression of the reporter, suggesting that HuR may be involved in the fine-tuning of the expression of the NF1 gene. PMID- 10673360 TI - Implication of novel biochemical property of beta-amyloid. AB - Alzheimer disease (AD) is a heterogeneous disorder with a variety of molecular pathologies converging predominantly on abnormal amyloid deposition particularly in the brain. beta-Amyloid aggregation into senile plaques is one of the pathological hallmarks of AD. beta-Amyloid is generated by a proteolytic cleavage of a large membrane protein, amyloid precursor protein (APP). We have observed a new property of beta-amyloid. The amyloid 1-42 beta fragment, when aggregated, possesses proteolytic and esterase-like activity, in vitro. Three independent methods were used to test the new property of beta-amyloid. While esterase activity involves imidazole catalysis, proteolytic activity is consistent with participation of a serine peptidase triad: catalytic Ser, His and Glu (or Asp). Although the amino acid triad is a necessary requirement for the protease reactivity, it is not sufficient since the secondary structure of the protein significantly contributes to the proteolytic activity. The ability of beta amyloid to cleave peptide or ester bonds could be thus responsible for either inactivation of other proteins and/or APP proteolysis itself. This property may be responsible for early pathogenesis of AD since there is emerging evidence that non-plaque amyloid is elevated in Alzheimer patients. PMID- 10673361 TI - Differential signaling pathways following oxidative stress in mutant myotonin protein kinase cDNA-transfected C2C12 cell lines. AB - We investigated the response to oxidative stress in a model system established in C2C12 cells stably transfected with myotonin protein kinase (MtPK) cDNAs having 5, 46, 60, or 160 CTG repeats. The transformants showed CTG repeat number dependent susceptibility to oxidative stress. Mutant MtPK cDNA transformants containing 160 CTG repeats showed apoptotic cell death by the exposure to an oxidant, a very low level of methylmercury. The addition of the antioxidant Trolox protected transformants against apoptosis. Oxidative stress activated the extracellular signal-regulated kinases (ERKs) pathway leading to cell survival in wild-type MtPK cDNA transformants, whereas mutant MtPK cDNA transformants having 160 CTG repeats were defective in the induction of the ERK pathway, although the activation of stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK) was strong and sustained. These results suggest that the susceptibility to oxidative stress in mutant MtPK cDNA transformants involves differential signaling pathways evoked following oxidative stress. PMID- 10673362 TI - Localization of exercise- and denervation-responsive elements in the mouse GLUT4 gene. AB - Exercise training increases the expression of GLUT4 in skeletal muscle. Previous studies demonstrated that the exercise-responsive element(s) of the murine GLUT4 gene are located between bases -1001 and -442 relative to the transcription start site. To further characterize the regulatory elements in the GLUT4 gene, the regulation of GLUT4 minigenes containing -701, -551, -442, or -423 bp of the 5' flanking region was studied in transgenic mice. All minigenes studied showed significant expression in skeletal muscle and heart, including the -423 GLUT4 minigene that lacked the myocyte enhancer factor 2 (MEF2)-binding domain ( CTAAAAATAG-) located between bases -437 and -428. The -701- and -551-bp constructs were expressed in brown adipose tissues while the -442 and -423 constructs were not. In skeletal muscle, either swimming or treadmill running up regulated GLUT4 minigene mRNA levels in -701 and -551 transgenic mice, but not in the -442 and -423 transgenic mice. Denervation of the gastrocnemius muscle by sectioning of the sciatic nerve down-regulated minigene and endogenous GLUT4 mRNAs in all -701, -551, -442, and -423 transgenic mice. These data indicate that exercise-responsive element(s) and brown adipocyte specific element(s) are located within 109 bp between bases -551 and -442 of the GLUT4 gene, but that the cis-element for denervation-induced down-regulation of the GLUT4 gene is located downstream of base -423. Finally, the MEF2 binding site between bases -437 and 428 is not necessary for expression of GLUT4 in skeletal muscles and heart; the cis-element mediating this effect is also located downstream of base -423. PMID- 10673363 TI - Hyperleptinemia in female patients with ossification of spinal ligaments. AB - In order to examine the involvement of leptin in the ossification of spinal ligaments (OSL), the present study examined (i) serum levels of leptin and insulin in OSL patients and controls, (ii) serum leptin levels in children of OSL females with severe obesity, (iii) the expression of leptin receptor mRNA in human spinal ligaments, and (iv) effects of leptin on cultured human ligament cells. In the OSL females, serum leptin levels were significantly higher than those of the control females, and the levels were positively correlated to the serum insulin levels, while in the control females, there was a tendency of inverse correlation. The daughters of OSL females with severe obesity also had high serum leptin levels, although they had not developed OSL. The expression of leptin receptor mRNA was confirmed in the ligaments, but leptin did not influence the alkaline phosphatase activity nor procollagen type I carboxyl-terminal peptide content of the ligament cells. These findings suggest that leptin is involved genetically and indirectly with the pathogenesis of OSL in female patients. PMID- 10673364 TI - Quantitative analysis of constitutive and inducible CYPs mRNA expression in the HepG2 cell line using reverse transcription-competitive PCR. AB - Drug interactions which affect drug metabolism are of clinical importance. It is, however, difficult to estimate drug interactions in human from results obtained in animal experiments. In our previous study, we demonstrated that a combination of the HepG2 cell line and semiquantitative reverse transcription-PCR (RT-PCR) could be used to evaluate the degree of CYP3A mRNA induction by various drugs. Using an RT-competitive PCR (RT-cPCR) with beta-actin as the standard in this study, the constitutive and rifampicin (RFP)-induced expression of CYP3A4, CYP2C9, CYP2E1, and CYP1A2 mRNA in the HepG2 cells could be quantitatively and reproducibly determined. 120 h-treatment of HepG2 cells with 50 micromol/l RFP induced maximally 8.4- and 6.0-fold the expression of CYP3A4 and CYP2C9 mRNA, respectively, in comparison with untreated cells. On the other hand, mRNA level in CYP2E1 and CYP1A2 was not significantly changed by 50 micromol/l RFP after 24 to 120 h. To our knowledge, we report for the first time quantitative profiles of CYPs mRNA in HepG2 cells. This study demonstrates the efficiency of a combination of HepG2 cells and RT-cPCR in the evaluation of CYPs mRNA-induction by drugs. PMID- 10673365 TI - Variants of NOS1, NOS2, and NOS3 genes in asthmatics. AB - Nitric oxide (NO) gas concentrations are higher in expired air in asthmatics. NO is synthesized by three isoforms of NO synthase (NOS) encoded by three distinct genes, NOS1, NOS2, and NOS3. Genome-wide searches have identified linkages to asthma on chromosomes 7, 12, and 17 where these three genes are localized. No association study, however, has been reported to date. To test whether variants of NOS1, NOS2, and NOS3 relate to asthma, a genetic association study was conducted in a British population (n = 300). Intragenic microsatellite variants of NOS1 were significantly associated with asthma [odds ratio (OR) = 2.08, 95% CI: 1.20-3.57 (95% CI), P = 0.008 (Pc = 0.048)], but not with IgE levels. Neither NOS2 nor NOS3 variants showed any association with asthma nor IgE levels. These findings suggest that NOS1 variants may be a significant contributor to asthma in a British population. PMID- 10673366 TI - A regulated interaction between alpha5beta1 integrin and osteopontin. AB - The extracellular matrix protein osteopontin (OPN) interacts with a number of integrins, namely alphavbeta1, alphavbeta3, alphavbeta5, alpha9beta1, alpha8beta1, and alpha4beta1. We have investigated the interaction of alpha5beta1 integrin with OPN using K562 cells, which only express alpha5beta1. alpha5beta1 is in a low activation state in this cell line, but can be stimulated to a higher activation state by the phorbol ester TPA. Treating K562 wild-type cells (K562 WT) with TPA stimulated an interaction between alpha5beta1 and OPN. No interaction was seen in the absence of TPA. alpha5beta1 selectively interacted with a GST fusion protein of the N-terminal fragment of OPN (aa17-168), which is generated in vivo by thrombin cleavage of OPN. Expression of the alpha4 integrin in K562 cells (K562-alpha4beta1) stimulated alpha5beta1-dependent binding to aa17 168 in the absence of TPA, suggesting that alpha4beta1 activates alpha5beta1 in K562 cells. Adhesion via alpha5beta1 is mediated by the Arg-Gly-Asp (RGD) motif of OPN, as mutating this sequence to Arg-Ala-Asp (RAD) blocked binding of both cell types. These data demonstrate that thrombin cleavage regulates the adhesive properties of OPN and that alpha5beta1 integrin can interact with thrombin cleaved osteopontin when in a high activation state. PMID- 10673367 TI - Differential regulation of FGF-1 and -2 mitogenic activity is related to their kinetics of binding to heparan sulfate in MDA-MB-231 human breast cancer cells. AB - The growth of the malignant human mammary MDA-MB-231 cells is stimulated by fibroblast growth factor-1 (FGF-1) but not by FGF-2. When these cells are cultured in the presence of chlorate, an inhibitor of heparan sulfate (HS) sulfation, their proliferation is stimulated by both FGF-1 and FGF-2. We analyzed the interactions of FGF-1 and FGF-2 with HS purified from the cell layer and the culture medium of control and chlorate-treated MDA-MB-231 cells. The HS from the cell layer bound FGF-1 with faster association kinetics than the HS from the culture medium, and so had a higher affinity for FGF-1. Chlorate treatment had no significant effect on the FGF-1 binding kinetics of the HS. In contrast to FGF-1, chlorate treatment of the cells significantly altered the FGF-2 binding kinetics. The HS from untreated cells possessed two binding sites for FGF-2, one with fast association kinetics (k(ass) 470,000 to 610,000 M(-1) s(-1)) and a high affinity (K(d) 46 to 70 nM) and one with slower association kinetics (k(ass) 74,000 to 100,000 M(-1) s(-1)) and a lower affinity (K(d) 290 to 400 nM). HS from chlorate treated cells possessed just a single binding site for FGF-2 with fast association kinetics (k(ass) 270,000 to 290,000 M(-1) s(-1)) and a high affinity (K(d) 41 to 57 nM). These results show that there is a relationship between the binding kinetics of FGFs and their ability to stimulate cell growth. PMID- 10673368 TI - A novel method for expression and large-scale production of human brain l glutamate decarboxylase. AB - l-Glutamate decarboxylase (GAD; EC 4.1.1.15) is the rate-limiting enzyme involved in the synthesis of gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the mammalian brain. Imbalance in the conversion of glutamate to GABA has been implicated in a host of human diseases. Studies on the structure, function, and therapeutic use of GAD have been precluded by insufficient quantities of purified active enzyme. Here we report a novel methodology for the expression and large-scale production of enzymatically active, pure, recombinant human GAD65 and GAD67. This method circumvents the sequestering of expressed protein into insoluble inclusion bodies and reduces production of truncated proteins. The availability of sufficient quantities of purified HGAD65 and HGAD67 has allowed for the production of specific polyclonal antibodies that discriminate between the two isoforms. This methodology, in addition to providing key human brain enzymes, may be generally applicable to other systems. PMID- 10673369 TI - Marked increase in membranolytic selectivity of novel cyclic tachyplesins constrained with an antiparallel two-beta strand cystine knot framework. AB - We have developed a highly constrained 18-residue cyclic peptide template based on the antimicrobial peptide tachyplesin-1 that features an end-to-end peptide backbone and a cystine knot-like motif with three evenly spaced disulfide bonds to cross-brace the antiparallel beta-strands and to approximate an amphiphatic "beta-tile"-like structure. Six beta-tile analogs were prepared to correlate different topological patterns with membranolytic specificity. Their conformations and antimicrobial and hemolytic activities were compared with tachyplesin-1 and the recently discovered Rhesus monkey theta defensin (RTD) which contains similar beta-tile structural elements. The beta-tile peptides and RTD retained broad spectrum antimicrobial activities. In general, they were less active than tachyplesin-1 in 10 tested organisms but their activity increased under high-salt (100 mM NaCl) rather than in low-salt conditions. The beta-tile peptides are highly nontoxic to human erythrocytes with EC(25) ranging from 600 to 4000 microM. Collectively, our results show that the design of a highly rigid peptide template is useful for further analog study to dissociate antimicrobial activity from cytotoxicity which would be helpful in discovering clinical applications for peptide antibiotics. PMID- 10673370 TI - Cathepsin Q, a novel lysosomal cysteine protease highly expressed in placenta. AB - The complete nucleotide sequence of a novel cathepsin cDNA derived from rat placenta was determined and is termed cathepsin Q. The predicted protein of 343 amino acid is a member of the family C1A protease related to cathepsin L. Rat cathepsin Q and its mouse counterpart were found highly expressed in placenta, whereas no detectable levels were found in lung, spleen, heart, brain, kidney, thymus, testicle, liver, or embryonic tissues. It is predicted that cathepsin Q will differ in catalytic specificity to another placental-specific protease, cathepsin P, indicating that these enzymes will have unique proteolytic functions in extra-embryonic tissues. PMID- 10673371 TI - Human membrane type-2 matrix metalloproteinase is defective in cell-associated activation of progelatinase A. AB - Transfection of the mouse membrane type-2 matrix metalloproteinase (MT2-MMP) gene into COS-1 cells resulted in activation of progelatinase A; however, that of the human gene had no effect. Expression of human and mouse MT2-MMP chimeric proteins revealed the defect of human MT2-MMP which resides in the region between amino acid (aa) residues 155 and 271. Seven aa residues in this region were not conserved between human and mouse MT2-MMP. Substitution with the corresponding mouse residue, proline-183 to serine and glutamine-185 to aspartic acid, recovered cell-associated progelatinase A activation function. These residues are located in the insertion sequence-2 (IS-2), which was conserved in six clones of the human MT2-MMP gene from different sources, except that of proline-183 which was substituted with serine from HT1080 cells. These results indicate that human MT2-MMP is defective in cell-associated activation of progelatinase A, and this is attributed to IS-2. These findings emphasize the importance of IS-2 in MT2-MMP functionality. PMID- 10673372 TI - Up-regulation of endothelin-converting enzyme-1 in gastric mucosal inflammatory responses to Helicobacter pylori lipopolysaccharide. AB - Endothelin-1 (ET-1) is a vasoactive peptide produced from a biologically inactive big ET-1 by the action of endothelin-converting enzyme-1 (ECE-1). We investigated gastric mucosal expression of ECE-1 during a 10-day course of inflammatory responses associated with acute gastritis elicited by Helicobacter pylori lipopolysaccharide. The ECE-1 activity was associated with microsomal fraction and the level of its expression reflected the extent of mucosal inflammatory involvement. The histologic pattern of inflammation reached a maximum on the 4th day following the lipopolysaccharide and was accompanied by a 4.1-fold enhancement in the expression of ECE-1 activity and a significant elevation in ET 1 (3.1-fold), TNF-alpha (8.8-fold), and apoptosis (11.6-fold). A 41.5% decrease in the severity of mucosal inflammation by the 10th day following the lipopolysaccharide was reflected in a 62.3% reduction in the mucosal expression of ECE-1 and a decline in TNF-alpha, ET-1, and apoptosis. Thus, H. pylori infection causes up-regulation of gastric mucosal ECE-1 expression, which leads to the enhancement of ET-1 production, induction of TNF-alpha, and triggering the apoptotic events that exacerbate the inflammatory process. PMID- 10673373 TI - Molecular cloning of neonate/infant-specific pepsinogens from rat stomach mucosa and their expressional change during development. AB - To clarify the nature of rat neonate/infant-specific pepsinogens, we carried out their purification and molecular cloning. Prochymosin was found to be the major neonatal pepsinogen. The general proteolytic activity of its active form, chymosin, was, however, lower than those of pepsins A and C which are predominant in adult animals. Molecular cloning of rat prochymosin cDNA was achieved along with cDNA for another neonate-specific pepsinogen, pepsinogen F, although determination of pepsinogen F in neonatal gastric mucosa was unsuccessful, presumably due to its lack of proteolytic activity or different proteolytic specificity. Northern blot analysis confirmed that genes for prochymosin and pepsinogen F are expressed only at neonatal/infant stages and the switching of gene expression to that of pepsinogen C occurred at late infant stages. A phylogenetic tree based on nucleotide sequences showed clearly that pepsinogens fall into four major groups, namely prochymosin and pepsinogen F of the neonate/infant and pepsinogens A and C of adult animals. Although, to date, prochymosin and pepsinogen F were believed to be expressed in only a limited number of mammals, the present results suggest that they might be expressed at the neonatal/infant stage in a variety of mammals. PMID- 10673374 TI - Biochemical evidence for a 170-kilodalton, AF-2-dependent vitamin D receptor/retinoid X receptor coactivator that is highly expressed in osteoblasts. AB - Human vitamin D receptor (hVDR) fused to glutathione S-transferase was utilized to detect a VDR-interacting protein (VIP) of approximately 170 kDa. VIP(170) is expressed in osteoblast-like ROS 17/2.8 cells and, to a lesser extent, in COS-7 and HeLa cells. VIP(170) may be a coactivator because it interacts only with 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) ligand-bound hVDR and because a mutation (E420A) in the activation function-2 (AF-2) of hVDR abolishes both receptor mediated transactivation and VIP(170) binding. Unlike L254G hVDR, a heterodimerization mutant with an intact AF-2, the E420A mutant is only partially attenuated in its association with the retinoid X receptor (RXR) DNA-binding partner. Finally, the ability of overexpressed hVDR to squelch glucocorticoid receptor-mediated transactivation is lost in both the L254G and E420A mutants. These results suggest that several protein-protein interactions, including VDR association with RXR and VIP(170), are required for stabilization of a multimeric complex that transduces the signal for 1,25(OH)(2)D(3)-elicited transactivation. PMID- 10673375 TI - Tyrosine phosphorylation of myelin P(0) and its implication in signal transduction. AB - P(0), a major structural protein of peripheral myelin, belongs to the immunoglobulin superfamily. Sequence comparison of P(0) with PZR, a tyrosine phosphatase SHP-2 binding protein we recently cloned, revealed the presence of an immunoreceptor tyrosine-based inhibitory motif (ITIM) in the intracellular portion of the P(0) molecule. To study the role of this putative ITIM in signal transduction, we have expressed P(0) in HT-1080 and 293 cells. Stimulation of the transfected cells with pervanadate, a powerful inhibitor of tyrosine phosphatases, resulted in tyrosine phosphorylation of P(0) and its association with several tyrosine-phosphorylated proteins. Mutation of Y(220) embedded in the ITIM to phenylalanine abolished the tyrosine phosphorylation and the association. Tyrosine phosphorylation of P(0) and its association with other signaling proteins were also observed in pervanadate-treated RN22 Schwannoma cells, which express endogenous P(0). Furthermore, injection of pervanadate induced tyrosine phosphorylation of P(0) in peripheral nerves of newborn but not adult mice. The physiological importance of the ITIM in P(0) is implied by the fact that a naturally occurred P(0) mutant with a disrupted ITIM has a dominant role in causing Dejerine-Scotts syndrome. Taken together, P(0) is phosphorylated on Try(220). The presence of an ITIM in P(0) and its ability to mediate protein protein interaction through tyrosine phosphorylation indicate that P(0) is not merely a structural protein but may also be a crucial player in cell signaling. PMID- 10673376 TI - The effect of apoptosis inhibitors on experimental autoimmune encephalomyelitis: apoptosis as a regulatory factor. AB - The effect of apoptosis inhibitors on experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, was investigated by intraperitoneal or intracisternal administration of apoptosis inhibitors Ac-YVAD-cmk and zVAD-fmk. After onset of the disease, these agents had no suppressive effect on EAE and resulted in impaired recovery or earlier relapse. Histological examination revealed that administration of zVAD-fmk suppressed the apoptotic death of inflammatory cells in the central nervous system (CNS) of mice with EAE. The results indicated that the apoptotic elimination of infiltrated cells in the CNS might be one of the recovery mechanisms in EAE. PMID- 10673377 TI - Abundant expression of 150-kDa oxygen-regulated protein in mouse pancreatic beta cells is correlated with insulin secretion. AB - The 150-kDa oxygen-regulated protein (ORP150) is a member of glucose-regulated proteins (GRPs), which are induced by stressful conditions such as oxygen or glucose deprivation. Here we investigated the highly abundant expression of ORP150 in mouse pancreas and its relationship with insulin secretion. Immunohistochemical analysis revealed that ORP150 expression was restricted to islets, especially to beta cells. The beta cell-specific expression was also observed in a mouse insulinoma cell line, MIN6, which secretes insulin in response to increased glucose concentration. Furthermore, ORP150 in islets dramatically diminished by fasting, concomitant with reduction of the serum insulin level. These results strongly suggest the role for ORP150 in insulin secretion. PMID- 10673378 TI - Electric field-mediated DNA encapsulation into large liposomes. AB - Large, ethidium bromide-loaded liposomes electrically pulsed in the presence of externally added DNA display the bright fluorescence of DNA-ethidium bromide complexes. Sonication of these liposomes increases the fluorescence of trapped DNA-ethidium bromide complexes by no more than about 40%. These results are thus in agreement with a mechanism involving electropores for DNA uptake but do not support an alternative mechanism, invoking invagination and pinching-off of the lipid bilayer, through which internalized DNA is shielded from the liposome contents. PMID- 10673379 TI - Hydrophilic residues at the apical domain of GroEL contribute to GroES binding but attenuate polypeptide binding. AB - The GroES binding site at the apical domain of GroEL, mostly consisting of hydrophobic residues, overlaps largely with the substrate polypeptide binding site. Essential contribution of hydrophobic interaction to the binding of both GroES and polypeptide was exemplified by the mutant GroEL(L237Q) which lost the ability to bind either of them. The binding site, however, contains three hydrophilic residues, E238, T261, and N265. For GroES binding, N265 is essential since GroEL(N265A) is unable to bind GroES. E238 contributes to rapid GroES binding to GroEL because GroEL(E238A) is extremely sluggish in GroES binding. Polypeptide binding was not impaired by any mutations of E238A, T261A, and N265A. Rather, these mutants, especially GroEL(N265A), showed stronger polypeptide binding affinity than wild-type GroEL. Thus, these hydrophilic residues have a dual role; they help GroES binding on one hand but attenuate polypeptide binding on the other hand. PMID- 10673380 TI - Structure-based development of pyridoxal propionate derivatives as specific inhibitors of cathepsin K in vitro and in vivo. AB - We found that pyridoxal phosphate shows considerable inhibition of cathepsins. CLIK-071, in which the phosphate ester of position 3 of pyridoxal phosphate was replaced by propionate, strongly inhibited cathepsin B. Three new types of synthetic pyridoxal propionate derivatives showing specific inhibition of cathepsin K were developed. New synthetic pyridoxal propionate derivatives, -162, -163, and -164, in which the methyl arm of position 6 of CLIK-071 was additionally modified, strongly inhibited cathepsin K and cathepsin S weakly, but other cathepsins were not inhibited. CLIK-166, in which the position 4 aldehyde of CLIK-071 is replaced by a vinyl radical and position 5 is additionally modified, showed cathepsin K-specific inhibition at 10(-5) M. Pit formation due to bone collagen degradation by cathepsin K of rat osteoclasts was specifically suppressed by administration of CLIK-164, but not by inhibitors of cathepsin L or B. PMID- 10673381 TI - The C1orf9 gene encodes a putative transmembrane member of a novel protein family. AB - Here we report the characterization of a human mRNA encoding a novel protein denoted C1orf9 (chromosome 1 open reading frame 9). The cDNA sequence, derived from a testis cDNA library, contains 5700 bp which encodes an open reading frame of 1254 amino acids. The deduced protein contains a putative N-terminal signal peptide and one putative transmembrane region, indicating membrane localization. No significant homology was found with known characterized proteins. However, a 150 amino acid region has significant homology to deduced protein sequences from other organisms, including Caenorhabditis elegans (43% identity), Saccharomyces cerevisiae (47% identity), Schizosaccharomyces pombe (48% identity), and two proteins from Arabidopsis thaliana (42% and 40% identity), suggesting a novel family of conserved domains. The C1orf9 gene was assigned to chromosome 1q24. The gene spans approximately 78.7 kb and is organized into at least 24 exons. Expression analysis revealed a single C1orf9 mRNA species of approximately 6.0 kb with a predominant expression in pancreas and testis, and only low levels of expression in other tissues examined. PMID- 10673382 TI - Rapid gene repression triggered by interleukin-6 at the onset of monocyte differentiation. AB - To date, the majority of characterized extracellular ligand-induced rapid changes in gene expression involve upregulation. Hence, rapid gene repression is either less common or less well studied. To study rapid gene repression during cytokine initiated differentiation programs, we used the mRNA subtractive hybridization technique of representational difference analysis to isolate repressed genes. Cultures of the myeloid leukemia cell line M1 were induced to terminally differentiate by treatment with interleukin-6 (IL-6). The repressed genes identified in our subtraction products include the genes encoding the growth factor receptor Flt3/Flk2/STK-1 (CD135) and the costimulatory protein CD24 [heat stable antigen] and the c-myb oncogene. Following 4 h of IL-6 treatment, mRNA levels of these genes are decreased by 45-65% relative to controls and after 8 h by 65-80%. Lipopolysaccharide also triggers the repression of these genes. Protein synthesis inhibitors do not block the IL-6-stimulated repression of c myb, or c-myc, mRNA, yet they do block the repression of flt3 and CD24 mRNA, demonstrating the existence of both protein synthesis-independent and -dependent mechanisms of cytokine-triggered rapid gene repression during differentiation. PMID- 10673383 TI - Coordinate recruitment of E-cadherin and ALCAM to cell-cell contacts by alpha catenin. AB - Here we report on the role of alpha-catenin in the cellular localization of activated leukocyte cell adhesion molecule, ALCAM, and cadherin-mediated cell adhesion in human prostate cancer cells. Cell lines that have a functional E cadherin-mediated cell adhesion (DU-145 and LNCaP) show ALCAM staining at cell cell contacts. In contrast, in cell lines that lack alpha-catenin expression (ALVA-31, PC-3, and PPC-1), E-cadherin-mediated adhesion is disturbed and ALCAM staining is cytoplasmic. A role of alpha-catenin in the recruitment of E-cadherin and ALCAM to cell-cell contacts was established by transfection of an alpha-N catenin construct into cell lines ALVA-31 and PC-3. This resulted not only in the correct assembly of E-cadherin/alpha-catenin complexes at the cell membrane but also in localization of ALCAM to cell-cell contacts, indicating that indeed alpha catenin affects ALCAM localization. PMID- 10673384 TI - Molecular mechanisms of platelet exocytosis: requirements for alpha-granule release. AB - Platelets function by secreting components necessary for primary clot formation. This report describes an in vitro assay that measures alpha-granule secretion. Using permeabilized platelets, it is possible to recreate Ca(2+)-stimulated release of platelet factor 4 (PF4) that is ATP- and temperature-dependent. Though other divalent cations can replace Ca(2+) (i.e., Sr(2+), Mn(2+), Zn(2+)), there is no effect of Ba(2+). Analysis by electron microscopy indicates that the in vitro assay also mimics the cytoskeletal rearrangements and granule centralization that occurs upon platelet activation in vivo. Antibody inhibition studies show that PF4 release requires the general membrane fusion protein N ethylmaleimide-sensitive factor (NSF) and well as the target membrane SNAP receptors (t-SNAREs), syntaxin 2, 4, and SNAP-23. As shown by electron microscopy, the anti-t-SNARE antibodies block granule to target membrane fusion. This finding is unique in that it is the first report of a role for two syntaxins in the same exocytosis event. PMID- 10673385 TI - The internalization and endosomal trafficking of the EGF receptor in response to EGF is delayed in the waved-2 mouse liver. AB - The EGF receptor in waved-2 mice contains a point mutation that renders it kinase deficient. We investigated how the waved-2 mutation affects the internalization and endosomal trafficking of the receptor in vivo in response to EGF. When the waved-2 mice were injected with EGF, there was approximately 50% less tyrosine phosphorylation detected in whole-liver homogenate compared to wild-type background mice. Although EGF increased the EGF receptor levels in the early and late liver endosomal fractions of waved-2 mice, its trafficking was delayed compared to wild-type mice. Ubiquitination of the EGF receptor may affect its endosomal sorting. We found that a similar amount of EGF receptor was immunoprecipitated from the endosomal fractions of EGF-treated waved-2 and wild type with anti-ubiquitin antibody. These results demonstrate that the waved-2 EGF receptor can become ubiquitinated and can be trafficked to the late endosomes, although it appears that its kinase deficiency delays this process. PMID- 10673386 TI - Cloning and characterization of an alpha-neurotoxin-type protein specific for the coral snake Micrurus corallinus. AB - During the cloning of abundant cDNAs expressed in the Micrurus corallinus coral snake venom gland, we cloned an alpha-neurotoxin homologue cDNA (nxh1). Two others isoforms were also cloned (nxh3 and nxh7, respectively). The nxh1 cDNA codes for a potential coral snake toxin with a signal peptide of 21 amino acids plus a predicted mature peptide with 57 amino acids. The deduced protein is highly similar to known toxic three-finger alpha-neurotoxins, with four deduced S S bridges at the same conserved positions. This is the first cDNA coding for a three-finger related protein described so far for coral snakes. However, the predicted protein does not possess some of the important amino acids for the nicotinic acetylcholine receptor interaction. This protein was expressed in Escherichia coli as a His-tagged protein that allowed the rapid purification of the recombinant protein. This protein was used to generate antibodies which recognized the recombinant protein in Western blot and also a single band present in the M. corallinus venom, but not in the venom of 10 other Micrurus species. PMID- 10673387 TI - Involvement of extracellular signal-regulated protein kinase in gliosis induced during recovery from metabolic inhibition. AB - Brain reperfusion may be of particular importance in the etiology of periventricular leukomalacia, of which the common findings are gliosis and ventricular dilatation. To investigate the mechanism of this pathogenesis, we used a metabolic inhibition (MI) model using cyanide plus deoxyglucose treatment of cultured glia isolated from fetal rat brain and examined the activity of extracellular signal-regulated protein kinase (ERK) during MI and also during the recovery from MI of 30 min. ERK activation was stimulated during MI and the recovery from MI. The time course and extent of activation of ERK during MI and the recovery from MI, however, were distinctly different. Activation of ERK was stimulated within 5 min of MI and declined thereafter. Activation of ERK was sustained during the recovery phase from MI and the extent of the activation was much greater than that during MI. Pretreatment with EGTA to eliminate extracellular Ca(2+), or with APV, an NMDA receptor antagonist, to inhibit Ca(2+) influx through the NMDA receptor, attenuated the activation of ERK. Moreover, pretreatment with PMA to downregulate PKC abolished the activation of ERK. PD98059, an inhibitor of ERK kinase, attenuated the cell proliferation induced by MI followed by recovery from MI. These results suggest that ERK is involved in gliosis during the recovery phase from MI and may play a role in the etiology of periventricular leukomalacia. PMID- 10673388 TI - The lipid component of lipoproteins from Borrelia burgdorferi: structural analysis, antigenicity, and presentation via human dendritic cells. AB - The spirochaetal bacteria Borrelia burgdorferi (Bb) is the tick-borne causative agent of lyme disease. The major membrane immunogens of Bb are outer surface proteins. The lipid component of these lipoproteins is relevant for the immunogenicity of Bb-lipoproteins. To characterize the antigenic properties, the native lipid component of lipoproteins was isolated and the detailed molecular structure was analyzed. The molecular structure of the lipoprotein-lipid component turned out to be S(propane-2',-3'diol)-3-thio-2-aminopropanic acid (S glyceryl-cysteine) with one ester-linked fatty acid, one acetyl group, and one N terminal amide-bound fatty acid. Fatty acid analysis of the lipid component indicated a heterogeneous composition comprising C16:0, C18:0, C18:1, C18:2, and C 20:0. The antigenicity was tested with in vitro bioassays using human blood derived dendritic cells (DCs) as antigen-presenting cells and autologous Bb specific T-cells. We found that human DCs present the lipid component of Bb lipoproteins via MHC class II inducing an antigen-specific T-cell immune response in vitro. PMID- 10673389 TI - BERP, a novel ring finger protein, binds to alpha-actinin-4. AB - We recently identified BERP as a novel RING finger protein belonging to the RBCC protein family. It contains an N-terminal RING finger, followed by a B-box zinc finger and a coiled-coil domain. BERP interacts with the tail domain of the class V myosins through a beta-propeller structure in the BERP C-terminal. To identify other proteins interacting with BERP, the yeast two-hybrid strategy was employed, using the RBCC domain as bait. Screening of a rat brain cDNA library identified alpha-actinin-4 as a specific binding partner for the N-terminus of BERP. This actinin isoform could be immunoprecipitated together with BERP from HEK 293 cells transfected with expression constructs for BERP and alpha-actinin-4. These proteins could also be colocalized immunohistochemically in the cytoplasm of differentiated PC12 cells. We suggest that BERP may anchor class V myosins to particular cell domains via its interaction with alpha-actinin-4. PMID- 10673390 TI - Role of troponin I isoform switching in determining the pH sensitivity of Ca(2+) regulation in developing rabbit cardiac muscle. AB - Skinned muscle fibers prepared from fetal rabbit heart (28 days of gestation) showed a marked resistance to acidic pH in the Ca(2+) regulation of force generation, compared to the fibers prepared from adult heart. SDS-PAGE and immunoblot analysis showed that the slow skeletal troponin I was predominantly expressed in the fetal cardiac muscle, while the cardiac isoform was predominantly expressed in the adult cardiac muscle. Direct exchange of purified slow skeletal and cardiac troponin I isoforms into these skinned muscle fibers revealed that cardiac troponin I made the Ca(2+) regulation of contraction sensitive to acidic pH just as in the adult fibers, whereas slow skeletal troponin I made the Ca(2+) regulation of contraction resistant to acidic pH just as in the fetal fibers. These results demonstrate that the troponin I isoform switching accounts fully for the change in the pH dependence of Ca(2+) regulation of contraction in developmental cardiac muscle. PMID- 10673391 TI - Novel functions of human alpha(1)-protease inhibitor after S-nitrosylation: inhibition of cysteine protease and antibacterial activity. AB - alpha(1)-Protease inhibitor (alpha(1)PI), the most abundant serine protease inhibitor found in human plasma (at 30-60 microM), is a glycoprotein (53 kDa) having a single cysteine residue at position 232 (Cys(232)). We have found that Cys(232) of human alpha(1)PI was readily S-nitrosylated by nitric oxide (NO) without affecting inhibitory activity to trypsin or elastase. S-nitrosylated alpha(1)PI (S-NO-alpha(1)PI) not only retained inhibitory activity against these serine proteases, but also gained thiol protease inhibitory activity against a Streptococcus pyogenes protease; the parental alpha(1)PI did not have this activity. Furthermore, S-NO-alpha(1)PI exhibited bacteriostatic activity against Salmonella typhimurium at concentrations of 0.1-10 microM, which were 20- to 3000 fold stronger than those of the other NO-generating compounds or S-nitroso compounds such as S-nitrosoalbumin and S-nitrosoglutathione. NO appears to be transferred into the bacterial cells from S-NO-alpha(1)PI via transnitrosylation, as evidenced by electron spin resonance spectroscopy with an NO spin trap. Thus, we conclude that S-NO-alpha(1)PI may be generated from the reaction between alpha(1)PI and NO under inflammatory conditions, in which production of both is known to increase. As a result, new functions, i.e., antibacterial and thiol protease inhibitory activities of alpha(1)PI, were generated. PMID- 10673392 TI - Biological activity of human granulocyte colony stimulating factor with a modified C-terminus. AB - Granulocyte colony-stimulating factor (G-CSF) undergoes receptor-mediated internalization into target cells which are normally restricted to neutrophilic granulocytes and their committed progenitor cells, suggesting that it may be applicable as a myeloid cell-targeting vehicle. To test this notion, we constructed a cDNA encoding a human G-CSF/murine stem cell factor (mSCF) chimeric molecule in a mammalian expression vector and transfected NIH3T3 cells with this plasmid. The resulting chimeric cytokine consisted of the entire G-CSF sequences fused to Lys148 of mSCF. It can be released from the surface membrane of NIH3T3 transformants through proteolytic cleavage at Ala164 of mSCF. The culture media conditioned by a number of stable transformants, which were confirmed by an enzyme-linked immunosorbent assay (ELISA) to secrete an hG-CSF derivative, were examined for their ability to stimulate CFU-G-derived colony formation as well as the proliferation of G-CSF-dependent NFS-60 cells. The results indicated that this C-terminus modified version of hG-CSF is as potent as recombinant hG-CSF in both assays. PMID- 10673393 TI - Heterooligomer of type 1 and type 2 inositol 1, 4, 5-trisphosphate receptor expressed in rat liver membrane fraction exists as tetrameric complex. AB - Functional IP(3)-sensitive intracellular Ca(2+) release channel is considered to be a tetramer of IP(3)R. Heterooligomeric complexes composed of distinct types of IP(3)R have been reported, however, crucial evidences for them being tetramer have not appeared. Here we report that the heterooligomer composed of IP(3)R1 and IP(3)R2 also exists as tetramer. Cross-linked heterooligomer was immunoprecipitated with IP(3)R1-specific antibody and detected by agarose PAGE/Western blot analysis with IP(3)R2-specific antibody. Tetramer, trimer, dimer, and possibly monomer were detected. The trimer, dimer, and monomer were likely to be originated from the tetramer, since: (1) the immunoprecipitating antibody (IP(3)R1-specific) does not recognize IP(3)R2, therefore IP(3)R2 monomer itself could not have been immunoprecipitated; and (2) tetramer was the major native product of IP(3)R complex containing type 2 isoform in liver membrane fraction. Thus we conclude tetramer is the native form of heterooligomer composed of IP(3)R1 and IP(3)R2. PMID- 10673394 TI - Specific modulation of p53 binding to consensus sequence within supercoiled DNA by monoclonal antibodies. AB - Monoclonal antibodies (MAbs) were used to investigate the binding of insect cell expressed, wild-type human p53 protein to the consensus sequence (p53CON) in a 474-bp DNA fragment and to supercoiled (sc) DNAs with and without p53CON. Supershifting of p53-DNA complexes by MAbs in agarose gels was applied to studies of activation of p53 for sequence-specific binding within scDNA. C-terminal specific antibody Bp53-10.1 activated the sequence-specific binding of p53 to p53CON within pPGM1 scDNA but did not influence binding of p53 to pBluescript scDNA (not containing p53CON). Incubation of p53 with DO-1 prior to addition of Bp53-10.1 prevented activation of p53 and induced dissociation of a portion of pPGM1 scDNA from the sequence-specific immune complex; no such dissociation was observed if pPGM1 scDNA was replaced by the 474-bp p53CON-containing DNA fragment. PMID- 10673395 TI - Structure-function analysis of the 7B2 CT peptide. AB - Prohormone convertases play important roles in the proteolytic conversion of many protein precursors. The neuroendocrine protein 7B2 and its 31-residue carboxyl terminal (CT) peptide potently and specifically inhibit prohormone convertase 2 (PC2). We have analyzed the residues contributing to inhibition using N-terminal truncation and alanine scanning. Removal of more than 3 residues from the amino terminal end of CT1-18 resulted in a more than 190-fold drop in inhibitory activity, showing that most of the residues between 3 and 18 are required for inhibition. In agreement, an Ala scan indicated that only 4 residues could be replaced with Ala without losing mid-nanomolar inhibitory potency; in particular, Gln7, Gln9, and Asp12 could be Ala-substituted to yield peptides with a similar inhibitory potency to the starting peptide. The all-d-retro-inverso, all-l inverso, and all-d analogues of CT peptide were completely inactive, indicating that amino acid side chains and the CT peptide main chain interact with PC2. CT peptide inhibition could not be competitively blocked by preincubation with truncated CT peptide forms, supporting an absolute requirement for the Lys-Lys pair in initial binding of the CT peptide to the active site. PMID- 10673396 TI - CFU-GM-derived cells form osteoclasts at a very high efficiency. AB - The granulocyte-macrophage progenitor (CFU-GM) is a multipotent cell that can differentiate to osteoclasts (OCLs), macrophages, or granulocytes. However, the relative potential of CFU-GM to efficiently form OCLs is unknown. In this report we demonstrate that granulocyte-macrophage colony-forming unit (CFU-GM)-derived cells represent an easily obtainable highly purified source of human OCL precursors that form OCLs at very high efficiency (greater than 90%) when cultured with RANK ligand (RANKL), macrophage colony-stimulating factor (M-CSF), and dexamethasone. The OCLs that formed have high bone-resorbing activity and form multiple resorption lacunae per OCL on dentin slices. Similarly, murine marrow-derived CFU-GM also formed OCLs at a high efficiency (>80%) when treated with RANKL, M-CSF, and dexamethasone. In contrast, more committed macrophage colony-forming unit (CFU-M)-derived cells form few OCLs under these conditions. PMID- 10673397 TI - beta3-endonexin as a novel inhibitor of cyclin A-associated kinase. AB - Cyclin A is indispensable for S phase cell cycle progression and is suggested to be a crucial target of cell adhesion signals. In this study, we demonstrate that beta3-endonexin, a molecule known to associate with the integrin beta3 cytoplasmic domain, specifically binds cyclin A. Deletion of the amino-terminal 52-amino-acid residues including the cyclin-binding RxL motif abolishes the ability of beta3-endonexin to interact with cyclin A. In an in vitro kinase assay, beta3-endonexin inhibits pRB kinase activity associated with cyclin A-Cdk2 while leaving its histone H1 kinase activity unaffected. Coexpression of beta3 endonexin in yeast cells overcomes growth suppression caused by an activation of cyclin A-associated kinase. Our results indicate that beta3-endonexin is a novel cyclin A-binding molecule that regulates cyclin A-associated pRB kinase activity. PMID- 10673398 TI - The oxidative stress-sensitive yap1 null strain of Saccharomyces cerevisiae becomes resistant due to increased carotenoid levels upon the introduction of the Chlamydomonas reinhardtii cDNA, coding for the 60S ribosomal protein L10a. AB - The Saccharomyces cerevisiae yap1 null strain was transformed with a Chlamydomonas reinhardtii cDNA expression library. A 688-bp cDNA fragment, coding for the 60S ribosomal protein L10a (RPL10a), restored the capacity of the S. cerevisiae yap1 null strain to resist oxidative stress. The rpl10a gene is a single-copy gene in C. reinhardtii and encodes a constitutively produced 1.35-kb mRNA. The deduced 214-residue amino acid sequence was highly related with RPL10a proteins from eukarya (between 46.1 and 63.7% identity) and archaea (between 24.5 and 29.2% identity). Resistant transformants were pink, due to increased carotenoid levels, with the same chemical structure as torularhodin, the main carotenoid of the pink yeast Rhodotorula mucilaginosa. The pink transformants showed high resistance levels against H(2)O(2), paraquat, menadione, and UV light. Partial inhibition of the carotenoid synthesis by diphenylamine reduced the resistance levels, demonstrating the role of excess carotenoid synthesis in the resistance mechanism. PMID- 10673399 TI - Interferon-gamma mediates gene expression of IL-18 binding protein in nonleukocytic cells. AB - Interleukin-18 (IL-18) binding protein is a soluble decoy receptor for IL-18 which efficiently antagonizes biological functions of IL-18 in vitro and in vivo. Since regulation of IL-18 activity likely contributes to the pathogenesis of inflammatory diseases as well as malignancies, we investigated gene expression of IL-18 binding protein (IL-18BP) in different human cell systems, namely in the keratinocyte cell line HaCaT, in the colon carcinoma cell line DLD-1, and in primary renal mesangial cells. In unstimulated cells only minute amounts of mRNA coding for IL-18 binding protein were detectable. However, in all three cell types gene expression was markedly upregulated by interferon-gamma (IFN-gamma). IL-18 is recognized as a pivotal mediator of IFN-gamma production. Therefore, the present data imply that activity of IL-18 is modulated by a negative feedback mechanism which is mediated by IFN-gamma-induced IL-18 binding protein. PMID- 10673400 TI - Biasing in Gaussian Random Fields and Galaxy Correlations. AB - In this Letter, we show that in a Gaussian random field, the correlation length the typical size of correlated structures-does not change with biasing. We interpret the amplification of the correlation functions of subsets identified by different thresholds as being caused by the increasing sparseness of peaks over threshold. This clarifies a long-standing misconception in the literature. We also argue that this effect does not explain the observed increase of the amplitude of the correlation function xi(r) when galaxies of brighter luminosity or galaxy clusters of increasing richness are considered. PMID- 10673401 TI - A Definitive Optical Detection of a Supercluster at z approximately 0.91. AB - We present the results from a multiband optical imaging program that has definitively confirmed the existence of a supercluster at z approximately 0.91. Two massive clusters of galaxies, Cl 1604+4304 at z=0.897 and Cl 1604+4321 at z=0.924, were originally observed in the high-redshift cluster survey of Oke, Postman, & Lubin. They are separated by 4300 km s-1 in radial velocity and 17&arcmin; on the plane of the sky. Their physical and redshift proximity suggested a promising supercluster candidate. Deep BRi imaging of the region between the two clusters indicates a large population of red galaxies. This population forms a tight, red sequence in the color-magnitude diagram at (R i&parr0; approximately 1.4. The characteristic color is identical to that of the spectroscopically confirmed early-type galaxies in the two member clusters. The red galaxies are spread throughout the 5 h-1 Mpc region between Cl 1604+4304 and Cl 1604+4321. Their spatial distribution delineates the entire large-scale structure with high concentrations at the cluster centers. In addition, we detect a significant overdensity of red galaxies directly between Cl 1604+4304 and Cl 1604+4321 which is the signature of a third, rich cluster associated with this system. The strong sequence of red galaxies and their spatial distribution clearly indicate that we have discovered a supercluster at z approximately 0.91. PMID- 10673402 TI - A Chandra High-Resolution X-ray Image of Centaurus A. AB - We present first results from a Chandra X-Ray Observatory observation of the radio galaxy Centaurus A with the High-Resolution Camera. All previously reported major sources of X-ray emission including the bright nucleus, the jet, individual point sources, and diffuse emission are resolved or detected. The spatial resolution of this observation is better than 1&arcsec; in the center of the field of view and allows us to resolve X-ray features of this galaxy not previously seen. In particular, we resolve individual knots of emission in the inner jet and diffuse emission between the knots. All of the knots are diffuse at the 1&arcsec; level, and several exhibit complex spatial structure. We find the nucleus to be extended by a few tenths of an arcsecond. Our image also suggests the presence of an X-ray counterjet. Weak X-ray emission from the southwest radio lobe is also seen, and we detect 63 pointlike galactic sources (probably X-ray binaries and supernova remnants) above a luminosity limit of approximately 1.7x1037 ergs s-1. PMID- 10673403 TI - Implications of the X-Ray Variability for the Mass of MCG -6-30-15. AB - The bright Seyfert 1 galaxy MCG -6-30-15 shows large variability on a variety of timescales. We study the less, similar3 day timescale variability using a set of simultaneous archival observations that were obtained from the Rossi X-Ray Timing Explorer (RXTE) and the Advanced Satellite for Cosmology and Astrophysics (ASCA). The RXTE observations span nearly 106 s and indicate that the X-ray Fourier power spectral density has an rms variability of 16%, is flat from approximately 10-6 to 10-5 Hz, and then steepens into a power law proportional to f-alpha with alpha greater, similar1. A further steepening to alpha approximately 2 occurs between 10-4 and 10-3 Hz. The shape and rms amplitude are comparable to what has been observed in NGC 5548 and Cyg X-1, albeit with break frequencies that differ by a factor of 10-2 and 104, respectively. If the break frequencies are indicative of the central black hole mass, then this mass may be as low as 106 M middle dot in circle. An upper limit of approximately 2 ks for the relative lag between the 0.5 2 keV ASCA band compared to the 8-15 keV RXTE band was also found. Again by analogy with NGC 5548 and Cyg X-1, this limit is consistent with a relatively low central black hole mass. PMID- 10673404 TI - A Unified Scaling Law in Spiral Galaxies. AB - We investigate the origin of a unified scaling relation in spiral galaxies. Observed spiral galaxies are spread on a plane in the three-dimensional logarithmic space of luminosity L, radius R, and rotation velocity V. The plane is expressed as L~&parl0;VR&parr0;alpha in the I passband, where alpha is a constant. On the plane, observed galaxies are distributed in an elongated region which looks like the shape of a surfboard. The well-known scaling relations L-V (Tully-Fisher [TF] relation), V-R (also the TF relation), and R-L (Freeman's law) can be understood as oblique projections of the surfboard-like plane into two dimensional spaces. This unified interpretation of the known scaling relations should be a clue to understand the physical origin of all the relations consistently. Furthermore, this interpretation can also explain why previous studies could not find any correlation between TF residuals and radius. In order to clarify the origin of this plane, we simulate formation and evolution of spiral galaxies with the N-body/smoothed particle hydrodynamics method, including cooling, star formation, and stellar feedback. Initial conditions are set to 14 isolated spheres with two free parameters, such as mass and angular momentum. The cold dark matter (h=0.5, Omega0=1) cosmology is considered as a test case. The simulations provide the following two conclusions: (1) The slope of the plane is well reproduced but the zero point is not. This zero-point discrepancy could be solved in a low-density (Omega0<1) and high-expansion (h>0.5) cosmology. (2) The surfboard-shaped plane can be explained by the control of galactic mass and angular momentum. PMID- 10673405 TI - The Modified Dynamics is Conducive to Galactic Warp Formation. AB - There is an effect in the modified dynamics that is conducive to the formation of warps. Because of the nonlinearity of the theory, the internal dynamics of a galaxy is affected by a perturber over and above possible tidal effects. For example, a relatively distant and light companion or the mean influence of a parent cluster, with negligible tidal effects, could still produce a significant warp in the outer part of a galactic disk. We present results of numerical calculations for simplified models that show, for instance, that a satellite with the (baryonic) mass and distance of the Magellanic Clouds can distort the axisymmetric field of the Milky Way enough to produce a warp of the magnitude (and position) observed. Details of the warp geometry remain to be explained; we use a static configuration that can produce only warps with a straight line of nodes. In more realistic simulations, one must reckon with the motion of the perturbing body, which sometimes occurs on timescales not much longer than the response time of the disk. PMID- 10673406 TI - The Optical Gravitational Lensing Experiment: Red Clump Stars as a Distance Indicator. AB - We present relation of the mean I-band brightness of red clump stars on metallicity. Red clump stars were proposed to be a very attractive standard candle for distance determination. The calibration is based on 284 nearby red giant stars whose high-quality spectra made it possible to determine accurate individual metal abundances. High-quality parallaxes (sigmapi&solm0;pi<10%) and photometry of these very bright stars come from Hipparcos measurements. Metallicity of the sample covers a large range: -0.6 dex<&sqbl0;Fe&solm0;H&sqbr0;<+0.2 dex. We find a weak dependence of the mean I band brightness on metallicity ( approximately 0.13 mag dex-1). What is more important, the range of metallicity of the Hipparcos sample partially overlaps with metallicity of field giants in the LMC, thus making it possible to determine the distance to the LMC by almost direct comparison of brightness of the local Hipparcos red clump giants with that of LMC stars. Photometry of field red clump giants in nine low-extinction fields of the LMC halo collected during the OGLE II microlensing survey compared with the Hipparcos red clump stars data yields the distance modulus to the LMC: &parl0;m-M&parr0;LMC=18.24+/-0.08 mag. PMID- 10673407 TI - Testing Population Synthesis Models with Globular Cluster Colors. AB - We have measured an extensive set of UBVRIJHK colors for M31 globular clusters. We compare the predicted simple stellar population colors of three population synthesis models to the intrinsic colors of Galactic and M31 globular clusters. The best-fitting models fit the cluster colors very well: the weighted mean color offsets are all smaller than 0.05 mag. The most significant offsets between model and data are in the U and B passbands; these are not unexpected and are likely caused by problems with the spectral libraries used by the models. The metal-rich clusters (&sqbl0;Fe&solm0;H&sqbr0; greater, similar-0.8) are best fit by young (8 Gyr) models, while the metal-poor clusters are best fit by older (12-16 Gyr) models. If this range of globular cluster ages is correct, it implies that conditions for cluster formation must have existed for a substantial fraction of the galaxies' lifetimes. PMID- 10673408 TI - Probing the Site for r-Process Nucleosynthesis with Abundances of Barium and Magnesium in Extremely Metal-poor Stars. AB - We suggest that if the astrophysical site for r-process nucleosynthesis in the early Galaxy is confined to a narrow mass range of Type II supernova (SN II) progenitors, with a lower mass limit of Mms=20 M middle dot in circle, a unique feature in the observed distribution of [Ba/Mg] versus [Mg/H] for extremely metal poor stars can be adequately reproduced. We associate this feature, a bifurcation of the observed elemental ratios into two branches in the Mg abundance interval 3.7/=10,000 K, however, are slowly rotating, vsini<7 km s-1, and their iron and titanium are enhanced by a factor of 300 to solar abundance levels. Magnesium maintains a nearly constant abundance over the entire range of Teff, and helium is depleted by factors of 10-30 in three of the hotter stars. Diffusion effects in the stellar atmospheres are the most likely explanation for these large differences in composition. Our results are qualitatively very similar to those previously reported for M13 and NGC 6752, but with even larger enhancement amplitudes, presumably due to the increased efficiency of radiative levitation at lower intrinsic [Fe/H]. We also see evidence for faster stellar rotation explicitly preventing the onset of the diffusion mechanisms among a subset of the hotter stars. PMID- 10673410 TI - Correlation among Quasi-Periodic Oscillation Frequencies and Quiescent-State Duration in Black Hole Candidate GRS 1915+105. AB - We discover a definite correlation between the frequency of the quasi-periodic oscillations (QPOs) in quiescent states and the duration of the quiescent state of the transient X-ray source GRS 1915+105. We find that while the QPO frequency can be explained by the oscillation of shocks in accretion flows, the switching of burst to quiescent states (and vice versa) and their duration can be explained by assuming an outflow from the postshock region. The duration of the quiescent state is inversely related to the QPO frequency. We derive this relation. We also find the correlation between the observed low ( approximately 0.001-0.01 Hz) and the intermediate (1-10 Hz) QPO frequencies. Our analytical solutions are verified by analyzing several days of public domain data from the Rossi X-Ray Timing Explorer. PMID- 10673411 TI - Phase Lag and Coherence Function of X-Ray Emission from Black Hole Candidate XTE J1550-564. AB - We report the results from measuring the phase lag and coherence function of X ray emission from black hole candidate XTE J1550-564. These temporal X-ray properties have been recognized to be increasingly important in providing important diagnostics of the dynamics of accretion flows around black holes. For XTE J1550-564, we found significant hard lag-the X-ray variability in high-energy bands lags behind that in low-energy bands-associated both with broadband variability and quasi-periodic oscillation (QPO). However, the situation is more complicated for the QPO: while hard lag was measured for the first harmonic of the signal, the fundamental component showed significant soft lag. Such behavior is remarkably similar to what was observed of microquasar GRS 1915+105. The phase lag evolved during the initial rising phase of the 1998 outburst. The magnitude of both the soft and hard lags of the QPO increases with X-ray flux, while the Fourier spectrum of the broadband lag varies significantly in shape. The coherence function is relatively high and roughly constant at low frequencies and begins to drop almost right after the first harmonic of the QPO. It is near unity at the beginning and decreases rapidly during the rising phase. Also observed is that the more widely separated the two energy bands are, the less the coherence function between the two. It is interesting that the coherence function increases significantly at the frequencies of the QPO and its harmonics. We discuss the implications of the results on the models proposed for black hole candidates. PMID- 10673412 TI - Advection-dominated Inflow/Outflows from Evaporating Accretion Disks. AB - In this Letter we investigate the properties of advection-dominated accretion flows (ADAFs) fed by the evaporation of a Shakura-Sunyaev accretion disk (SSD). In our picture, the ADAF fills the central cavity evacuated by the SSD and extends beyond the transition radius into a coronal region. We find that, because of global angular momentum conservation, a significant fraction of the hot gas flows away from the black hole, forming a transsonic wind, unless the injection rate depends only weakly on radius (if r2sigma&d2;~r-xi, xi<1&solm0;2). The Bernoulli number of the inflowing gas is negative if the transition radius is less, similar100 Schwarzschild radii, so matter falling into the hole is gravitationally bound. The ratio of inflowing to outflowing mass is approximately 1/2, so in these solutions the accretion rate is of the same order as in standard ADAFs and much larger than in advection-dominated inflow/outflow models. The possible relevance of evaporation-fed solutions to accretion flows in black hole X-ray binaries is briefly discussed. PMID- 10673413 TI - The Compact Central Object in Cassiopeia A: A Neutron Star with Hot Polar Caps or a Black Hole? AB - The central pointlike X-ray source of the Cassiopeia A supernova remnant was discovered in the Chandra first light observation and found later in the archival ROSAT and Einstein images. The analysis of these data does not show statistically significant variability of the source. Because of the small number of photons detected, different spectral models can fit the observed spectrum. The power-law fit yields the photon index gamma=2.6-4.1, and luminosity L(0.1-5.0 keV&parr0;=&parl0;2-60&parr0;x1034 ergs s-1 for d=3.4 kpc. The power-law index is higher, and the luminosity lower, than those observed from very young pulsars. One can fit the spectrum equally well with a blackbody model with T=6-8 MK, R=0.2 0.5 km, and Lbol=&parl0;1.4-1.9&parr0;x1033 ergs s-1. The inferred radii are too small, and the temperatures too high, for the radiation to be interpreted as emitted from the whole surface of a uniformly heated neutron star. Fits with the neutron star atmosphere models increase the radius and reduce the temperature, but these parameters are still substantially different from those expected for a young neutron star. One cannot exclude, however, the possibility that the observed emission originates from hot spots on a cooler neutron star surface. An upper limit on the (gravitationally redshifted) surface temperature is Tinfinitys<1.9-2.3 MK, depending on the chemical composition of the surface and the star's radius. Among several possible interpretations, we favor a model of a strongly magnetized neutron star with magnetically confined hydrogen or helium polar caps (Tinfinitypc approximately 2.8 MK, Rpc approximately 1 km) on a cooler iron surface (Tinfinitys approximately 1.7 MK). Such temperatures are consistent with the standard models of neutron star cooling. Alternatively, the observed radiation may be interpreted as emitted by a compact object (more likely, a black hole) accreting from a residual disk or from a late-type dwarf in a close binary. PMID- 10673414 TI - Discovery of a Brown Dwarf Companion to Gliese 570ABC: A 2MASS T Dwarf Significantly Cooler than Gliese 229B. AB - We report the discovery of a widely separated (258&farcs;3+/-0&farcs;4) T dwarf companion to the Gl 570ABC system. This new component, Gl 570D, was initially identified from the Two Micron All-Sky Survey. Its near-infrared spectrum shows the 1.6 and 2.2 um CH4 absorption bands characteristic of T dwarfs, while its common proper motion with the Gl 570ABC system confirms companionship. Gl 570D (MJ=16.47+/-0.07) is nearly a full magnitude dimmer than the only other known T dwarf companion, Gl 229B, and estimates of L=&parl0;2.8+/-0.3&parr0;x10-6 L middle dot in circle and Teff=750+/-50 K make it significantly cooler and less luminous than any other known brown dwarf companion. Using evolutionary models by Burrows et al. and an adopted age of 2-10 Gyr, we derive a mass estimate of 50+/ 20 MJup for this object. PMID- 10673415 TI - The Discovery of a Second Field Methane Brown Dwarf from Sloan Digital Sky Survey Commissioning Data. AB - We report the discovery of a second field methane brown dwarf from the commissioning data of the Sloan Digital Sky Survey (SDSS). The object, SDSS J134646.45-003150.4 (hereafter SDSS 1346-00), was selected because of its very red color and stellar appearance. Its spectrum between 0.8 and 2.5 um is dominated by strong absorption bands of H2O and CH4 and closely mimics those of Gliese 229B and SDSS 162414.37+002915.6 (hereafter SDSS 1624+00), two other known methane brown dwarfs. SDSS 1346-00 is approximately 1.5 mag fainter than Gliese 229B, suggesting that it lies about 11 pc from the Sun. The ratio of flux at 2.1 um to that at 1.27 um is larger for SDSS 1346-00 than for Gliese 229B and SDSS 1624+00, which suggests that SDSS 1346-00 has a slightly higher effective temperature than the others. Based on a search area of 130 deg2 and a detection limit of z*=19.8, we estimate a space density of 0.05 pc-3 for methane brown dwarfs with Teff approximately 1000 K in the 40 pc3 volume of our search. This estimate is based on small-sample statistics and should be treated with appropriate caution. PMID- 10673416 TI - Analysis of the Hipparcos Measurements of HD 10697: A Mass Determination of a Brown Dwarf Secondary. AB - HD 10697 is a nearby main-sequence star around which a planet candidate has recently been discovered by means of radial velocity measurements (Vogt et al.). The stellar orbit has a period of about 3 yr, the secondary minimum mass is 6.35 Jupiter masses (MJ), and the minimum semimajor axis is 0.36 mas. Using the Hipparcos data of HD 10697 together with the spectroscopic elements of Vogt et al., we found a semimajor axis of 2.1+/-0.7 mas, implying a mass of 38+/-13 MJ for the unseen companion. We therefore suggest that the secondary of HD 10697 is probably a brown dwarf, orbiting around its parent star at a distance of 2 AU. PMID- 10673417 TI - Ultraviolet Photoprocessing of Interstellar Dust Mantles as a Source of Polycyclic Aromatic Hydrocarbons and Other Conjugated Molecules. AB - By co-depositing a gas mixture of simple carbon- and nitrogen-containing molecules with water on a 10 K surface and exposing it to ultraviolet radiation, we were able to form a residue. This residue was then placed aboard the EURECA satellite behind a magnesium fluoride window and exposed to solar radiation for 4 months before it was returned and analyzed. The resulting residue is believed to simulate the photoprocessing of organic dust mantles in the interstellar medium. Mass spectrometry indicated that the photoprocessing created a rich mixture of polycyclic aromatic hydrocarbons (PAHs) and other conjugated organic molecules, which may explain how PAHs are replenished in space. PMID- 10673418 TI - Implosions in Coronal Transients. AB - Coronal events such as flares or coronal mass ejections derive their energy from the energy stored locally in the magnetic field. This leads to the conjecture that a magnetic implosion must occur simultaneously with the energy release. The site of the implosion would show the location of preflare energy storage, and its detection should have a high priority. The Transition Region and Coronal Explorer EUV observations, for example, have sufficient resolution to show the geometry of a flare implosion by following the motions of tracers in the images. PMID- 10673419 TI - Identification of the Coronal Sources of the Fast Solar Wind. AB - The present spectroscopic study of the ultraviolet coronal emission in a polar hole, detected on 1996 April 6-9 with the Ultraviolet Coronagraph Spectrometer aboard the Solar and Heliospheric Observatory spacecraft, identifies the interplume lanes and background coronal hole regions as the channels in which the fast solar wind is preferentially accelerated. In interplume lanes, at heliocentric distance 1.7 R middle dot in circle, the corona expands at a rate between 105 and 150 km s-1, that is, much faster than in plumes in which the outflow velocity is between 0 and 65 km s-1. The wind velocity is inferred from the Doppler dimming of the O vi lambdalambda1032, 1037 lines, within a range of values, whose lower and upper limit corresponds to anisotropic and isotropic velocity distribution of the oxygen coronal ions, respectively. PMID- 10673420 TI - Initial Time Dependence of Abundances in Solar Energetic Particle Events. AB - We compare the initial behavior of Fe/O and He/H abundance ratios and their relationship to the evolution of the proton energy spectra in "small" and "large" gradual solar energetic particle (SEP) events. The results are qualitatively consistent with the behavior predicted by the theory of Ng et al. published in 1999. He/H ratios that initially rise with time are a signature of scattering by non-Kolmogorov Alfven wave spectra generated by intense beams of shock accelerated protons streaming outward in large gradual SEP events. PMID- 10673421 TI - Crystal structure of RNA 3'-terminal phosphate cyclase, a ubiquitous enzyme with unusual topology. AB - BACKGROUND: RNA cyclases are a family of RNA-modifying enzymes that are conserved in eucarya, bacteria and archaea. They catalyze the ATP-dependent conversion of the 3'-phosphate to the 2',3'-cyclic phosphodiester at the end of RNA, in a reaction involving formation of the covalent AMP-cyclase intermediate. These enzymes might be responsible for production of the cyclic phosphate RNA ends that are known to be required by many RNA ligases in both prokaryotes and eukaryotes. RESULTS: The high-resolution structure of the Escherichia coli RNA 3'-terminal phosphate cyclase was determined using multiwavelength anomalous diffraction. Two orthorhombic crystal forms of E. coli cyclase (space group P2(1)2(1)2(1) and P2(1)2(1)2) were used to solve and refine the structure to 2.1 A resolution (R factor 20.4%; R(free) 27.6%). Each molecule of RNA cyclase consists of two domains. The larger domain contains three repeats of a folding unit comprising two parallel alpha helices and a four-stranded beta sheet; this fold was previously identified in translation initiation factor 3 (IF3). The large domain is similar to one of the two domains of 5-enolpyruvylshikimate-3-phosphate synthase and UDP-N-acetylglucosamine enolpyruvyl transferase. The smaller domain uses a similar secondary structure element with different topology, observed in many other proteins such as thioredoxin. CONCLUSIONS: The fold of RNA cyclase consists of known elements connected in a new and unique manner. Although the active site of this enzyme could not be unambiguously assigned, it can be mapped to a region surrounding His309, an adenylate acceptor, in which a number of amino acids are highly conserved in the enzyme from different sources. The structure of E. coli cyclase will be useful for interpretation of structural and mechanistic features of this and other related enzymes. PMID- 10673422 TI - The crystal structure of the formiminotransferase domain of formiminotransferase cyclodeaminase: implications for substrate channeling in a bifunctional enzyme. AB - BACKGROUND: The bifunctional enzyme formiminotransferase-cyclodeaminase (FTCD) contains two active sites at different positions on the protein structure. The enzyme binds a gamma-linked polyglutamylated form of the tetrahydrofolate substrate and channels the product of the transferase reaction from the transferase active site to the cyclodeaminase active site. Structural studies of this bifunctional enzyme and its monofunctional domains will provide insight into the mechanism of substrate channeling and the two catalytic reactions. RESULTS: The crystal structure of the formiminotransferase (FT) domain of FTCD has been determined in the presence of a product analog, folinic acid. The overall structure shows that the FT domain comprises two subdomains that adopt a novel alpha/beta fold. Inspection of the folinic acid binding site reveals an electrostatic tunnel traversing the width of the molecule. The distribution of charged residues in the tunnel provides insight into the possible mode of substrate binding and channeling. The electron density reveals that the non natural stereoisomer, (6R)-folinic acid, binds to the protein; this observation suggests a mechanism for product release. In addition, a single molecule of glycerol is bound to the enzyme and indicates a putative binding site for formiminoglutamate. CONCLUSIONS: The structure of the FT domain in the presence of folinic acid reveals a possible novel mechanism for substrate channeling. The position of the folinic acid and a bound glycerol molecule near to the sidechain of His82 suggests that this residue may act as the catalytic base required for the formiminotransferase mechanism. PMID- 10673423 TI - The high-resolution structure of the NADP(H)-binding component (dIII) of proton translocating transhydrogenase from human heart mitochondria. AB - BACKGROUND: Transhydrogenase, located in the inner membranes of animal mitochondria and the cytoplasmic membranes of bacteria, couples the transfer of reducing equivalents between NAD(H) and NADP(H) to proton pumping. The protein comprises three subunits termed dI, dII and dIII. The dII component spans the membrane. The dI component, which contains the binding site for NAD(+)/NADH, and the dIII component, which has the binding site for NADP(+)/NADPH, protrude from the membrane. Proton pumping is probably coupled to changes in the binding affinities of dIII for NADP(+) and NADPH. RESULTS: The first X-ray structure of the NADP(H)-binding component, dIII, of human heart transhydrogenase is described here at 2.0 A resolution. It comprises a single domain resembling the classical Rossmann fold, but NADP(+) binds to dIII with a reversed orientation. The first betaalphabetaalphabeta motif of dIII contains a Gly-X-Gly-X-X-Ala/Val 'fingerprint', but it has a different function to that in the classical Rossmann structure. The nicotinamide ring of NADP(+) is located on a ridge where it is exposed to interaction with NADH on the dI subunit. Two distinctive features of the dIII structure are helix D/loop D, which projects from the beta sheet, and loop E, which forms a 'lid' over the bound NADP(+). CONCLUSIONS: Helix D/loop D interacts with the bound nucleotide and loop E, and probably interacts with the membrane-spanning dII. Changes in ionisation and conformation in helix D/loop D, resulting from proton translocation through dII, are thought to be responsible for the changes in affinity of dIII for NADP(+) and NADPH that drive the reaction. PMID- 10673424 TI - Mapping the binding site for the GTP-binding protein Rac-1 on its inhibitor RhoGDI-1. AB - BACKGROUND: Members of the Rho family of small GTP-binding proteins, such as Rho, Rac and Cdc42, have a role in a wide range of cell responses. These proteins function as molecular switches by virtue of a conformational change between the GTP-bound (active) and GDP-bound (inactive) forms. In addition, most members of the Rho and Rac subfamilies cycle between the cytosol and membrane. The cytosolic guanine nucleotide dissociation inhibitors, RhoGDIs, regulate both the GDP/GTP exchange cycle and the membrane association/dissociation cycle. RESULTS: We have used NMR spectroscopy and site-directed mutagenesis to identify the regions of human RhoGDI-1 that are involved in binding Rac-1. The results emphasise the importance of the flexible regions of both proteins in the interaction. At least one specific region (residues 46-57) of the flexible N-terminal domain of RhoGDI, which has a tendency to form an amphipathic helix in the free protein, makes a major contribution to the binding energy of the complex. In addition, the primary site of Rac-1 binding on the folded domain of RhoGDI involves the beta4-beta5 and beta6-beta7 loops, with a slight movement of the 3(10) helix accompanying the interaction. This binding site is on the same face of the protein as the binding site for the isoprenyl group of post-translationally modified Rac-1, but is distinct from this site. CONCLUSIONS: Isoprenylated Rac-1 appears to interact with three distinct sites on RhoGDI. The isoprenyl group attached to the C terminus of Rac-1 binds in a pocket in the folded domain of RhoGDI. This is distinct from the major site on this domain occupied by Rac-1 itself, which involves two loops at the opposite end to the isoprenyl-binding site. It is probable that the flexible C-terminal region of Rac-1 extends from the site at which Rac-1 contacts the folded domain of RhoGDI to allow the isoprenyl group to bind in the pocket at the other end of the RhoGDI molecule. Finally, the flexible N terminus of RhoGDI-1, and particularly residues 48-58, makes a specific interaction with Rac-1 which contributes substantially to the binding affinity. PMID- 10673425 TI - Novel fold and assembly of the repetitive B region of the Staphylococcus aureus collagen-binding surface protein. AB - BACKGROUND: [corrected] The Staphylococcus aureus collagen-binding protein Cna mediates bacterial adherence to collagen. The primary sequence of Cna has a non repetitive collagen-binding A region, followed by the repetitive B region. The B region has one to four 23 kDa repeat units (B(1)-B(4)), depending on the strain of origin. The affinity of the A region for collagen is independent of the B region. However, the B repeat units have been suggested to serve as a 'stalk' that projects the A region from the bacterial surface and thus facilitate bacterial adherence to collagen. To understand the biological role of these B region repeats we determined their three-dimensional structure. RESULTS: B(1) has two domains (D(1) and D(2)) placed side-by-side. D(1) and D(2) have similar secondary structure and exhibit a unique fold that resembles but is the inverse of the immunoglobulin-like (IgG-like) domains. Comparison with similar immunoglobulin superfamily (IgSF) structures shows novel packing arrangements between the D(1) and D(2) domains. In the B(1)B(2) crystal structure, an omission of a single glycine residue in the D(2)-D(3) linker loop, compared to the D(1) D(2) and D(3)-D(4) linker loops, resulted in projection of the D(3) and D(4) in a spatially new orientation. We also present a model for B(1)B(2)B(3)B(4). CONCLUSIONS: The B region of the Cna collagen adhesin has a novel fold that is reminiscent of but is inverse in nature to the IgG fold. This B region assembly could effectively provide the needed flexibility and stability for presenting the ligand binding A region away from the bacterial cell surface. PMID- 10673426 TI - The structure of TolB, an essential component of the tol-dependent translocation system, and its protein-protein interaction with the translocation domain of colicin E9. AB - BACKGROUND: E colicin proteins have three functional domains, each of which is implicated in one of the stages of killing Escherichia coli cells: receptor binding, translocation and cytotoxicity. The central (R) domain is responsible for receptor-binding activity whereas the N-terminal (T) domain mediates translocation, the process by which the C-terminal cytotoxic domain is transported from the receptor to the site of its cytotoxicity. The translocation of enzymatic E colicins like colicin E9 is dependent upon TolB but the details of the process are not known. RESULTS: We have demonstrated a protein-protein interaction between the T domain of colicin E9 and TolB, an essential component of the tol-dependent translocation system in E. coli, using the yeast two-hybrid system. The crystal structure of TolB, a procaryotic tryptophan-aspartate (WD) repeat protein, reveals an N-terminal alpha + beta domain based on a five stranded mixed beta sheet and a C-terminal six-bladed beta-propeller domain. CONCLUSIONS: The results suggest that the TolB-box residues of the T domain of colicin E9 interact with the beta-propeller domain of TolB. The protein-protein interactions of other beta-propeller-containing proteins, the yeast yPrp4 protein and G proteins, are mediated by the loops or outer sheets of the propeller blades. The determination of the three-dimensional structure of the T domain-TolB complex and the isolation of mutations in TolB that abolish the interaction with the T domain will reveal fine details of the protein-protein interaction of TolB and the T domain of E colicins. PMID- 10673427 TI - Characterization of the unfolding pathway of the cell-cycle protein p13suc1 by molecular dynamics simulations: implications for domain swapping. AB - BACKGROUND: The p13suc1 gene product is a member of the cks (cyclin-dependent protein kinase subunit) protein family and has been implicated in regulation of the cell cycle. Various crystal structures of suc1 are available, including a globular, monomeric form and a beta-strand exchanged dimer. It has been suggested that conversions between these forms, and perhaps others, may be important in the regulation of the cell cycle. RESULTS: We have undertaken molecular dynamics simulations of protein unfolding to investigate the conformational properties of suc1. Unfolding transition states were identified for each of four simulations. These states contain some native secondary structure, primarily helix alpha1 and the core of the beta sheet. The hydrophobic core is loosely packed. Further unfolding leads to an intermediate state that is slightly more expanded than the transition state, but with considerably fewer nonlocal, tertiary packing contacts and less secondary structure. The helices are fluctuating but partially formed in the denatured state and beta2 and beta4 remain associated. CONCLUSIONS: It appears that suc1 folds by a nucleation-condensation mechanism, similar to that observed for two-state folding proteins. However, suc1 forms an intermediate during unfolding and contains considerable residual structure in the denatured state. The stability of the beta2-beta4 residual structure is surprising, because beta4 is the strand involved in domain swapping. This stability suggests that the domain-swapping event, if physiologically relevant, may require the assistance of additional factors in vivo or occur early in the folding process. PMID- 10673428 TI - A new scaffold for binding haem in the cytochrome domain of the extracellular flavocytochrome cellobiose dehydrogenase. AB - BACKGROUND: The fungal oxidoreductase cellobiose dehydrogenase (CDH) degrades both lignin and cellulose, and is the only known extracellular flavocytochrome. This haemoflavoenzyme has a multidomain organisation with a b-type cytochrome domain linked to a large flavodehydrogenase domain. The two domains can be separated proteolytically to yield a functional cytochrome and a flavodehydrogenase. Here, we report the crystal structure of the cytochrome domain of CDH. RESULTS: The crystal structure of the b-type cytochrome domain of CDH from the wood-degrading fungus Phanerochaete chrysosporium has been determined at 1.9 A resolution using multiple isomorphous replacement including anomalous scattering information. Three models of the cytochrome have been refined: the in vitro prepared cytochrome in its redox-inactive state (pH 7.5) and redox-active state (pH 4.6), as well as the naturally occurring cytochrome fragment. CONCLUSIONS: The 190-residue long cytochrome domain of CDH folds as a beta sandwich with the topology of the antibody Fab V(H) domain. The haem iron is ligated by Met65 and His163, which confirms previous results from spectroscopic studies. This is only the second example of a b-type cytochrome with this ligation, the first being cytochrome b(562). The haem-propionate groups are surface exposed and, therefore, might play a role in the association between the cytochrome and flavoprotein domain, and in interdomain electron transfer. There are no large differences in overall structure of the cytochrome at redox-active pH as compared with the inactive form, which excludes the possibility that pH dependent redox inactivation results from partial denaturation. From the electron density map of the naturally occurring cytochrome, we conclude that it corresponds to the proteolytically prepared cytochrome domain. PMID- 10673429 TI - Structures of human dihydroorotate dehydrogenase in complex with antiproliferative agents. AB - BACKGROUND: Dihydroorotate dehydrogenase (DHODH) catalyzes the fourth committed step in the de novo biosynthesis of pyrimidines. As rapidly proliferating human T cells have an exceptional requirement for de novo pyrimidine biosynthesis, small molecule DHODH inhibitors constitute an attractive therapeutic approach to autoimmune diseases, immunosuppression, and cancer. Neither the structure of human DHODH nor any member of its family was known. RESULTS: The high-resolution crystal structures of human DHODH in complex with two different inhibitors have been solved. The initial set of phases was obtained using multiwavelength anomalous diffraction phasing with selenomethionine-containing DHODH. The structures have been refined to crystallographic R factors of 16.8% and 16.2% at resolutions of 1. 6 A and 1.8 A for inhibitors related to brequinar and leflunomide, respectively. CONCLUSIONS: Human DHODH has two domains: an alpha/beta-barrel domain containing the active site and an alpha-helical domain that forms the opening of a tunnel leading to the active site. Both inhibitors share a common binding site in this tunnel, and differences in the binding region govern drug sensitivity or resistance. The active site of human DHODH is generally similar to that of the previously reported bacterial active site. The greatest differences are that the catalytic base removing the proton from dihydroorotate is a serine rather than a cysteine, and that packing of the flavin mononucleotide in its binding site is tighter. PMID- 10673430 TI - The manifold of vitamin B6 dependent enzymes. AB - Pyridoxal-5'-phosphate (vitamin B6) binding enzymes form a large superfamily that contains at least five different folds. The availability of an increasing number of known three-dimensional structures for members of this superfamily has allowed a detailed structural classification. Most progress has been made with the fold type I or aspartate aminotransferase family. PMID- 10673431 TI - The folding pathway of the cell-cycle regulatory protein p13suc1: clues for the mechanism of domain swapping. AB - BACKGROUND: The 113-residue alpha+beta protein suc1 is a member of the cyclin dependent kinase subunit (cks) family of proteins that are involved in regulation of the eukaryotic cell cycle. In vitro, suc1 undergoes domain swapping to form a dimer by the exchange of a C-terminal beta strand. We have analysed the folding pathway of suc1 in order to determine the atomic details of how strand-exchange occurs in vitro and thereby obtain clues as to the possible mechanism and functional role of dimerisation in vivo. RESULTS: The structures of the rate determining transition state for the folding/unfolding of suc1 and of the intermediate that is populated during refolding were probed using phi values determined for 57 mutants with substitutions at 43 sites throughout the protein. The majority of phi values are fractional in the intermediate and transition state, indicating that interactions build up in a concerted manner during folding. In the transition state, phi values of greater than 0.5 are clustered around the inner strands beta2 and beta4 of the beta sheet. This part of the structure constitutes the nucleus for folding according to a nucleation condensation mechanism. Molecular dynamics simulations of unfolding of suc1, performed independently in a blind manner, are in excellent agreement with experiment (proceeding paper). CONCLUSIONS: Strand beta4 is the exchanging strand in the dimer and yet it forms an integral part of the folding nucleus. This suggests that association is an early event in the folding reaction of the dimer. Therefore, interchange between the monomer and dimer must occur via an unfolded state, a process that may be facilitated in vivo by accessory proteins. PMID- 10673432 TI - Structural and kinetic analysis of Escherichia coli GDP-mannose 4,6 dehydratase provides insights into the enzyme's catalytic mechanism and regulation by GDP fucose. AB - BACKGROUND: GDP-mannose 4,6 dehydratase (GMD) catalyzes the conversion of GDP-(D) mannose to GDP-4-keto, 6-deoxy-(D)-mannose. This is the first and regulatory step in the de novo biosynthesis of GDP-(L)-fucose. Fucose forms part of a number of glycoconjugates, including the ABO blood groups and the selectin ligand sialyl Lewis X. Defects in GDP-fucose metabolism have been linked to leukocyte adhesion deficiency type II (LADII). RESULTS: The structure of the GDP-mannose 4,6 dehydratase apo enzyme has been determined and refined using data to 2.3 A resolution. GMD is a homodimeric protein with each monomer composed of two domains. The larger N-terminal domain binds the NADP(H) cofactor in a classical Rossmann fold and the C-terminal domain harbors the sugar-nucleotide binding site. We have determined the GMD dissociation constants for NADP, NADPH and GDP mannose. Each GMD monomer binds one cofactor and one substrate molecule, suggesting that both subunits are catalytically competent. GDP-fucose acts as a competitive inhibitor, suggesting that it binds to the same site as GDP-mannose, providing a mechanism for the feedback inhibition of fucose biosynthesis. CONCLUSIONS: The X-ray structure of GMD reveals that it is a member of the short chain dehydrogenase/reductase (SDR) family of proteins. We have modeled the binding of NADP and GDP-mannose to the enzyme and mutated four of the active-site residues to determine their function. The combined modeling and mutagenesis data suggests that at position 133 threonine substitutes serine as part of the serine tyrosine-lysine catalytic triad common to the SDR family and Glu 135 functions as an active-site base. PMID- 10673433 TI - The in situ conformation and axial location of the intermolecular cross-linked non-helical telopeptides of type I collagen. AB - BACKGROUND: Type I collagen contains specific lysine and hydroxylysine residues that are critical in the formation of intermolecular cross-links crucial for the normal configuration and stability of the 67 nm axial repeat of collagen fibrils in the extracellular matrix. The major cross-linkage sites are believed to occur between the non-helical terminal regions (telopeptides) and helical segments of adjacent collagen molecules. In this X-ray fibre diffraction study the tissue has been maintained in the hydrated fibrillar state, whilst detailed structural information was obtained using highly collimated synchrotron radiation. RESULTS: The axial component of the X-ray diffraction patterns extends more than twice as far in reciprocal space than that of any already published. The structure-factor phases were calculated using the multiple isomorphous addition method, avoiding model-based approaches, and produced an electron-density profile of the molecular arrangement projected on to the fibre axis to 0.54 nm resolution. This corresponds to the phasing of 124 orders of the meridional diffraction pattern. CONCLUSIONS: The axially projected electron-density profile and the electron density difference maps showed that both the N- and C-terminal telopeptides are contracted structures. This profile puts narrow constraints on the possible conformations of the C-terminal telopeptide; the best fit to the electron-density profile is when the alpha1 chains adopt a folded conformation with a sharp hairpin turn around residues 13 and 14 of the 25-residue telopeptide. Our results reveal for the first time the location, parallel to the fibril axis, of the intermolecular cross-links in normal hydrated tissue. These cross-links are essential for the biological function of the tissue. PMID- 10673434 TI - The basis for K-Ras4B binding specificity to protein farnesyltransferase revealed by 2 A resolution ternary complex structures. AB - BACKGROUND: The protein farnesyltransferase (FTase) catalyzes addition of the hydrophobic farnesyl isoprenoid to a cysteine residue fourth from the C terminus of several protein acceptors that are essential for cellular signal transduction such as Ras and Rho. This addition is necessary for the biological function of the modified proteins. The majority of Ras-related human cancers are associated with oncogenic variants of K-RasB, which is the highest affinity natural substrate of FTase. Inhibition of FTase causes regression of Ras-mediated tumors in animal models. RESULTS: We present four ternary complexes of rat FTase co crystallized with farnesyl diphosphate analogs and K-Ras4B peptide substrates. The Ca(1)a(2)X portion of the peptide substrate binds in an extended conformation in the hydrophobic cavity of FTase and coordinates the active site zinc ion. These complexes offer the first view of the polybasic region of the K-Ras4B peptide substrate, which confers the major enhancement of affinity of this substrate. The polybasic region forms a type I beta turn and binds along the rim of the hydrophobic cavity. Removal of the catalytically essential zinc ion results in a dramatically different peptide conformation in which the Ca(1)a(2)X motif adopts a beta turn. A manganese ion binds to the diphosphate mimic of the farnesyl diphosphate analog. CONCLUSIONS: These ternary complexes provide new insight into the molecular basis of peptide substrate specificity, and further define the roles of zinc and magnesium in the prenyltransferase reaction. Zinc is essential for productive Ca(1)a(2)X peptide binding, suggesting that the beta turn conformation identified in previous nuclear magnetic resonance (NMR) studies reflects a state in which the cysteine is not coordinated to the zinc ion. The structural information presented here should facilitate structure-based design and optimization of inhibitors of Ca(1)a(2)X protein prenyltransferases. PMID- 10673435 TI - The 2.0 A crystal structure of Thermus thermophilus methionyl-tRNA synthetase reveals two RNA-binding modules. AB - BACKGROUND: The 20 aminoacyl-tRNA synthetases are divided into two classes, I and II. The 10 class I synthetases are considered to have in common the catalytic domain structure based on the Rossmann fold, which is totally different from the class II catalytic domain structure. The class I synthetases are further divided into three subclasses, a, b and c, according to sequence homology. No conserved structural features for tRNA recognition by class I synthetases have been established. RESULTS: We determined the crystal structure of the class Ia methionyl-tRNA synthetase (MetRS) at 2.0 A resolution, using MetRS from an extreme thermophile, Thermus thermophilus HB8. The T. thermophilus MetRS structure is in full agreement with the biochemical and genetic data from Escherichia coli MetRS. The conserved 'anticodon-binding' residues are spatially clustered on an alpha-helix-bundle domain. The Rossmann-fold and anticodon binding domains are connected by a beta-alpha-alpha-beta-alpha topology ('SC fold') domain that contains the class I specific KMSKS motif. CONCLUSIONS: The alpha-helix-bundle domain identified in the MetRS structure is the signature of the class Ia enzymes, as it was also identified in the class Ia structures of the isoleucyl- and arginyl-tRNA synthetases. The beta-alpha-alpha-beta-alpha topology domain, which can now be identified in all known structures of the class Ia and Ib synthetases, is likely to dock with the inner side of the L-shaped tRNA, thereby positioning the anticodon stem. PMID- 10673436 TI - Structural basis of the Ca(2+)-dependent association between S100C (S100A11) and its target, the N-terminal part of annexin I. AB - BACKGROUND: S100C (S100A11) is a member of the S100 calcium-binding protein family, the function of which is not yet entirely clear, but may include cytoskeleton assembly and dynamics. S100 proteins consist of two EF-hand calcium binding motifs, connected by a flexible loop. Like several other members of the family, S100C forms a homodimer. A number of S100 proteins form complexes with annexins, another family of calcium-binding proteins that also bind to phospholipids. Structural studies have been undertaken to understand the basis of these interactions. RESULTS: We have solved the crystal structure of a complex of calcium-loaded S100C with a synthetic peptide that corresponds to the first 14 residues of the annexin I N terminus at 2.3 A resolution. We find a stoichiometry of one peptide per S100C monomer, the entire complex structure consisting of two peptides per S100C dimer. Each peptide, however, interacts with both monomers of the S100C dimer. The two S100C molecules of the dimer are linked by a disulphide bridge. The structure is surprisingly close to that of the p11-annexin II N terminal peptide complex solved previously. We have performed competition experiments to try to understand the specificity of the S100-annexin interaction. CONCLUSIONS: By solving the structure of a second annexin N terminus-S100 protein complex, we confirmed a novel mode of interaction of S100 proteins with their target peptides; there is a one-to-one stoichiometry, where the dimeric structure of the S100 protein is, nevertheless, essential for complex formation. Our structure can provide a model for a Ca(2+)-regulated annexin I-S100C heterotetramer, possibly involved in crosslinking membrane surfaces or organising membranes during certain fusion events. PMID- 10673437 TI - The 1.8 A crystal structure and active-site architecture of beta-ketoacyl-acyl carrier protein synthase III (FabH) from escherichia coli. AB - BACKGROUND: beta-Ketoacyl-acyl carrier protein synthase III (FabH) initiates elongation in type II fatty acid synthase systems found in bacteria and plants. FabH is a ubiquitous component of the type II system and is positioned ideally in the pathway to control the production of fatty acids. The elucidation of the structure of FabH is important for the understanding of its regulation by feedback inhibition and its interaction with drugs. Although the structures of two related condensing enzymes are known, the roles of the active-site residues have not been experimentally tested. RESULTS: The 1.8 A crystal structure of FabH was determined using a 12-site selenium multiwavelength anomalous dispersion experiment. The active site (Cys112, His244 and Asn274) is formed by the convergence of two alpha helices and is accessed via a narrow hydrophobic tunnel. Hydrogen-bonding networks that include two tightly bound water molecules fix the positions of His244 and Asn274, which are critical for the decarboxylation and condensation reactions. Surprisingly, the His244-->Ala mutation does not affect the transacylation reaction suggesting that His244 has only a minor influence on the nucleophilicity of Cys112. CONCLUSIONS: The histidine and asparagine active site residues are both required for the decarboxylation step in the condensation reaction. The nucleophilicity of the active-site cysteine is enhanced by the alpha-helix dipole effect, and an oxyanion hole promotes the formation of the tetrahedral transition state. PMID- 10673438 TI - The structure of adenylosuccinate lyase, an enzyme with dual activity in the de novo purine biosynthetic pathway. AB - BACKGROUND: Adenylosuccinate lyase is an enzyme that plays a critical role in both cellular replication and metabolism via its action in the de novo purine biosynthetic pathway. Adenylosuccinate lyase is the only enzyme in this pathway to catalyze two separate reactions, enabling it to participate in the addition of a nitrogen at two different positions in adenosine monophosphate. Both reactions catalyzed by adenylosuccinate lyase involve the beta-elimination of fumarate. Enzymes that catalyze this type of reaction belong to a superfamily, the members of which are homotetramers. Because adenylosuccinate lyase plays an integral part in maintaining proper cellular metabolism, mutations in the human enzyme can have severe clinical consequences, including mental retardation with autistic features. RESULTS: The 1.8 A crystal structure of adenylosuccinate lyase from Thermotoga maritima has been determined by multiwavelength anomalous dispersion using the selenomethionine-substituted enzyme. The fold of the monomer is reminiscent of other members of the beta-elimination superfamily. However, its active tetrameric form exhibits striking differences in active-site architecture and cleft size. CONCLUSIONS: This first structure of an adenylosuccinate lyase reveals that, along with the catalytic base (His141) and the catalytic acid (His68), Gln212 and Asn270 might play a vital role in catalysis by properly orienting the succinyl moiety of the substrates. We propose a model for the dual activity of adenylosuccinate lyase: a single 180 degrees bond rotation must occur in the substrate between the first and second enzymatic reactions. Modeling of the pathogenic human S413P mutation indicates that the mutation destabilizes the enzyme by disrupting the C-terminal extension. PMID- 10673439 TI - Structure of Aspergillus niger epoxide hydrolase at 1.8 A resolution: implications for the structure and function of the mammalian microsomal class of epoxide hydrolases. AB - BACKGROUND: Epoxide hydrolases have important roles in the defense of cells against potentially harmful epoxides. Conversion of epoxides into less toxic and more easily excreted diols is a universally successful strategy. A number of microorganisms employ the same chemistry to process epoxides for use as carbon sources. RESULTS: The X-ray structure of the epoxide hydrolase from Aspergillus niger was determined at 3.5 A resolution using the multiwavelength anomalous dispersion (MAD) method, and then refined at 1.8 A resolution. There is a dimer consisting of two 44 kDa subunits in the asymmetric unit. Each subunit consists of an alpha/beta hydrolase fold, and a primarily helical lid over the active site. The dimer interface includes lid-lid interactions as well as contributions from an N-terminal meander. The active site contains a classical catalytic triad, and two tyrosines and a glutamic acid residue that are likely to assist in catalysis. CONCLUSIONS: The Aspergillus enzyme provides the first structure of an epoxide hydrolase with strong relationships to the most important enzyme of human epoxide metabolism, the microsomal epoxide hydrolase. Differences in active-site residues, especially in components that assist in epoxide ring opening and hydrolysis of the enzyme-substrate intermediate, might explain why the fungal enzyme attains the greater speeds necessary for an effective metabolic enzyme. The N-terminal domain that is characteristic of microsomal epoxide hydrolases corresponds to a meander that is critical for dimer formation in the Aspergillus enzyme. PMID- 10673440 TI - The atomic-resolution structure of a novel bacterial esterase. AB - BACKGROUND: A novel bacterial esterase that cleaves esters on halogenated cyclic compounds has been isolated from an Alcaligenes species. This esterase 713 is encoded by a 1062 base pair gene. The presence of a leader sequence of 27 amino acids suggests that this enzyme is exported from the cytosol. Esterase 713 has been over-expressed in Agrobacterium without this leader sequence. Its amino acid sequence shows no significant homology to any known protein sequence. RESULTS: The crystal structure of esterase 713 has been determined by multiple isomorphous replacement and refined to 1. 1 A resolution. The subunits of this dimeric enzyme comprise a single domain with an alpha/beta hydrolase fold. The catalytic triad has been identified as Ser206-His298-Glu230. The acidic residue of the catalytic triad (Glu230) is located on the beta6 strand of the alpha/beta hydrolase fold, whereas most other alpha/beta hydrolase enzymes have the acidic residue located on the beta7 strand. The oxyanion hole is formed by the mainchain nitrogens of Cys71 and Gln207 as identified by the binding of a substrate analogue, (S)-7-iodo 2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1, 4-benzodiazepine-2-acetic acid. Cys71 forms a disulphide bond with the neighbouring Cys72. CONCLUSIONS: Despite negligible sequence homology, esterase 713 has structural similarities to a number of other esterases and lipases. Residues of the oxyanion hole were confirmed by structural comparison with Rhizomucor miehei lipase. It is proposed that completion of a functional active site requires the formation of the disulphide bond between adjacent residues Cys71 and Cys72 on export of the esterase into the oxidising environment of the periplasmic space. PMID- 10673441 TI - Phospholipase A(2) enzymes: structural diversity in lipid messenger metabolism. AB - The phospholipases A(2) (PLA(2)s) are a large family of enzymes with varied lipidic products which are involved in numerous signal transduction pathways. The structural and functional characterization of several PLA(2)s have revealed the various mechanisms used by these enzymes to ingeniously manipulate the phospholipidic metabolic machinery. PMID- 10673442 TI - A new variant of the Ntn hydrolase fold revealed by the crystal structure of L aminopeptidase D-ala-esterase/amidase from Ochrobactrum anthropi. AB - BACKGROUND: The L-aminopeptidase D-Ala-esterase/amidase from Ochrobactrum anthropi (DmpA) releases the N-terminal L and/or D-Ala residues from peptide substrates. This is the only known enzyme to liberate N-terminal amino acids with both D and L stereospecificity. The DmpA active form is an alphabeta heterodimer, which results from a putative autocatalytic cleavage of an inactive precursor polypeptide. RESULTS: The crystal structure of the enzyme has been determined to 1.82 A resolution using the multiple isomorphous replacement method. The heterodimer folds into a single domain organised as an alphabetabetaalpha sandwich in which two mixed beta sheets are flanked on both sides by two alpha helices. CONCLUSIONS: DmpA shows no similarity to other known aminopeptidases in either fold or catalytic mechanism, and thus represents the first example of a novel family of aminopeptidases. The protein fold of DmpA does, however, show structural homology to members of the N-terminal nucleophile (Ntn) hydrolase superfamily. DmpA presents functionally equivalent residues in the catalytic centre when compared with other Ntn hydrolases, and is therefore likely to use the same catalytic mechanism. In spite of this homology, the direction and connectivity of the secondary structure elements differ significantly from the consensus Ntn hydrolase topology. The DmpA structure thus characterises a new subfamily, but supports the common catalytic mechanism for these enzymes suggesting an evolutionary relationship. PMID- 10673443 TI - Simple redox-linked proton-transfer design: new insights from structures of quinol-fumarate reductase. AB - The mitochondrial bioenergetics field has experienced an exciting breakthrough with the recent structure determination of several key membrane complexes. The latest addition to this line of structures, that of quinol-fumarate reductase, provides new insights into the mechanism of energy transduction. PMID- 10673444 TI - Giant solitary trichoepithelioma. AB - The giant solitary trichoepithelioma is a rare trichogenic tumor with potential for local recurrence. Only nine cases have been previously described in the literature, and one additional case without recurrence during the first 3.5 years of observation is presented stressing that the rate of recurrence is low. PMID- 10673445 TI - Pigmented purpuric dermatosis (Schamberg's purpura) in an infant. AB - Purpura in an infant is usually an alarming sign of a systemic (infectious, hematologic-oncologic or immunologic) disease. Chronic purpuric dermatoses have not been reported in infants and, therefore, are not considered in the differential diagnosis of purpuras in this age. We report on a female infant with progressive purpura who underwent extensive laboratory investigations to rule out a systemic disease. Based on the laboratory findings and clinical course, the diagnosis of Schamberg's purpura was established. PMID- 10673446 TI - Hereditary hypodontia and onychorrhexis of the fingernails and toenail koilonychia: Witkop's tooth-and nail syndrome. AB - The tooth-and-nail syndrome (Witkop's syndrome) is a rare autosomal dominant ectodermal dysplasia manifest by defects of the nail plates of the fingers and toes and hypodontia with normal hair and sweat gland function. We report a thirteen year-old girl who presented with marked longitudinal ridging of the nail plates of all ten fingers. The toenails were mildly ridged with koilonychia. Her mother's fingers were similarly affected to a lesser degree while her toenails appeared normal. Examination of the child's dentition revealed a hyperplastic frenulum and the absence of one of the usual four mandibular incisors. History provided by the mother described the maternal grandmother and maternal great aunt as having identical nail findings and the presence of only three lower incisors. Hair examination was normal in the mother and child, and no history or findings of sweat gland dysfunction was present. This report describes familial hypodontia, fingernail onychorrhexis, and toenail koilonychia consistent with Witkop's syndrome. PMID- 10673447 TI - Long-standing translucent tumor on the groin. Diagnosis: mucinous carcinoma of the skin. PMID- 10673448 TI - Benign lymphoepithelial tumor of the skin ("cutaneous lymphadenoma"). AB - "Benign lymphoepithelial tumor of the skin" or "cutaneous lymphadenoma" is a distinctive benign adnexal tumor presenting a characteristic combination of lobules of epithelial basaloid cells with a peculiar histopathological pattern and a dense intraepithelial T-cell lymphoid and histiocytic population. We report an additional example of this peculiar neoplasm. In addition to the characteristic histopathological features, focal areas showing unequivocal follicular differentiation were observed at the periphery of the tumor. In agreement with other authors we consider that this neoplasm should be included within the spectrum of neoplasms of pilosebaceous origin. Nevertheless, we consider that the original and simple concept of "benign lymphoepithelial tumor of the skin" seems more suitable and illustrative than the more widely used term of "cutaneous lymphadenoma" to define this rare benign adnexal neoplasm. PMID- 10673449 TI - Cimex lectularius. What is this insect and how does it affect man? AB - Cimex lectularius, the common bed bug, is a bloodsucking nocturnal parasite of man. Other hosts for this bug include chickens, bats, and some domestic animals. Cimex feeds by piercing the host with hollow tubes derived from the maxilla. Saliva injected at the time of feeding is associated with local and sometimes widespread urticaria. This pest has become less important over the last half century with general improvements in household and personal cleanliness. PMID- 10673450 TI - Delayed type hypersensitivity: current theories with an historic perspective. AB - Although the delayed type hypersensitivity (DTH) reaction was discovered over 100 years ago, the exact nature of the reaction has been the subject of contentious debate over the years. The reaction was discovered in 1882 by Robert Koch, but it was not until the 1940s that Landsteiner and Chase proved that the reaction was mediated by the cellular and not the humoral arm of the immune system. The first DTH reaction described used only the tuberculin antigen (tuberculin reaction), but the definition was later expanded to include cell mediated reactions to other bacterial and viral antigens, responses to pure protein with adjuvant or haptens, and host responses to allograft. The DTH skin test is used to test if prior exposure to an antigen has occurred. When small quantities of antigen are injected dermally, a hallmark response is elicited which includes induration, swelling and monocytic infiltration into the site of the lesion within 24 to 72 hours. This reaction has been shown to be absolutely dependent on the presence of memory T cells. Both the CD4+ and CD8+ fractions of cells have been shown to modulate a response. Contemporary debate regarding the reaction is focused on the role of the Th1 and Th2 cells originally discovered by Mosmann. It has been postulated that the Th1 cell is the "inducer" of a DTH response since it secretes interferon gamma (IFN ), a potent stimulator of macrophages, while the Th2 cell is either not involved or acting as a downregulator of the cell mediated immune response. Despite the early experimental success of this theory, experiments have shown that Th2 cells may be involved in certain types of proinflammatory cell mediated immunity. This review focuses on the nature of the different forms of DTH that can be elicited and the different experimental evidence that has led to the current theories regarding DTH and its role in cell mediated immunity. PMID- 10673451 TI - Sweet's syndrome (acute febrile neutrophilic dermatosis). AB - Sweet's syndrome, or acute febrile neutrophilic dermatosis, is a condition characterized by the sudden onset of fever, leukocytosis, and tender,erythematous, well-demarcated papules and plaques which show dense neutrophilic infiltrates on histologic examination. Although it ma occur in the absence of other known disease, Sweet's syndrome is often associated with hematologic disease (including leukemia), and immunologic disease (rheumatoid arthritis, inflammatory bowel disease). Treatment with systemic corticosteroids is usually successful. skin, Sweet syndrome, neutrophilic dermatosis, corticosteroids, PMID- 10673452 TI - Publishing dermatology therapy reviews on-line. PMID- 10673454 TI - Insulin-like growth factor 1 and hair growth. AB - Insulin-like growth factor 1 (IGF-1) has been identified as an important growth factor in many biological systems.[1] It shares considerable structural homology with insulin and exerts insulin-like effects on food intake and glucose metabolism. Recently it has been suggested to play a role in regulating cellular proliferation and migration during the development of hair follicles. [2,3] To exert its biological effects, the IGF-1 is required to activate cells by binding to specific cell-surface receptors. The type I IGF receptor (IGF-1R) is the only IGF receptor to have IGF-mediated signaling functions.[1] In circulation, this growth factor mediates endocrine action of growth hormone (GH) on somatic growth and is bound to specific binding proteins (BPs). The latter control IGF transport, efflux from vascular compartments and association with cell surface receptors.[4] In tissues, IGF-1 is produced by mesenchymal type cells and acts in a paracrine and autocrine fashion by binding to the IGF-1R. This binding activates the receptor tyrosine kinase (RTK) that triggers the downstream responses and finally stimulates cell division.[5] IGF-1 may therefore be able to stimulate the proliferation of hair follicle cells through cellular signaling pathways of its receptors. Local infusion of IGF-1 into sheep has been reported to be capable of stimulating protein synthesis in the skin.[6] It may also increase the production of wool keratin. Recently, transgenic mice overexpressing IGF-1 in the skin have been shown to have earlier hair follicle development than controls.[7] In addition, this growth factor plays an important role in many cell types as a survival factor to prevent cell death.[8] This anti-apoptotic function of IGF-1 may be important to the development of follicle cells as follicles undergo a growth cycle where the regressive, catagen phase is apoptosis driven. In this review, the effects of IGF-1 on follicle cell proliferation and differentiation are discussed. In particular, the paracrine versus endocrine action of IGF-1 on hair growth and the targeting of expression of the growth factor to the follicles of transgenic animals will be emphasized. The anti apoptotic role of IGF-1 in hair follicles is also reviewed. Prospects for future studies on hair and fiber growth by IGF-1 are discussed. PMID- 10673455 TI - Isotretinoin induced rhabdomyolysis? A case report. AB - Isotretinoin, an effective therapy for nodulocystic acne and dissecting cellulitis of the scalp, has many known side effects. However, its association with elevated creatine kinase levels and its potential to cause rhabdomyolysis is not well established. We describe a patient with a significant elevation in creatine kinase after beginning therapy with isotretinoin for dissecting cellulitis of the scalp. The implications of isotretinoin causing rhabdomyolysis are discussed. PMID- 10673456 TI - The clinical diagnosis of early malignant melanoma: expansion of the ABCD criteria to improve diagnostic sensitivity. AB - With the steady increase in incidence of malignant melanomas (MM) in the United States, early diagnosis and complete removal are critical for the containment of the malignancy. [1] The "ABCD" method of identification, originally described by Friedman et al., has been a useful tool in facilitating the diagnosis of MM. [2,3,4] This method analyzes four clinical characteristics to identify a malignant melanoma: Asymmetry, Border irregularity, Color variegation, and a Diameter of 6 mm or more.[4] Clinicians recognize that some melanomas lack all or most of the features defined in the "ABCD" rules. [5] This may be especially true of some early invasive and in situ melanomas. [6,7] In these instances, clinical history documenting morphologic change over time can be an important additional consideration. The following case reports underscore the need to expand the ABCD mnemonic to include an "E" for "Evolutionary change." An additional modification is also needed to emphasize the need for a low threshold for biopsy of unusual lesions which do not show typical benign features, even if they do not meet the ABCDE criteria. To this end we propose an "F" for "Funny looking lesions". PMID- 10673457 TI - The relationship between biopsy technique and uncertainty in the histopathologic diagnosis of melanoma. AB - Review of filed histopathology material of 525 cases of definite melanomas and other atypical pigmented lesions showed that diagnostic certainty was greatest for excisional or deep shave specimens. Shave and particularly punch biopsy specimens were associated with less certainty. The data of this study suggest that punch biopsy should be avoided for pigmented lesions and that a properly done deep shave biopsy is nearly equal to an excision in diagnostic content for such lesions. PMID- 10673458 TI - Mudi-chood disease. AB - Mudi-chood disease is a distinct clinical entity seen nearly exclusively in young women in Kerala State, India. Traditional hair grooming methods that utilize various plant oils along with the natural environmental heat and humidity create the conditions necessary to produce a lichenoid dermatitis on the neck and upper back. PMID- 10673459 TI - Genus Tabanus. Tabanids (horseflies). What is this insect and how does it affect man? AB - The insect family Tabanidae (Order: Diptera) comprises approximately 3000 species of flies worldwide, including the commonly known horseflies and deerflies. Females of most species require a blood meal to complete egg development and many species are important in human and animal medicine. This article briefly discusses the life cycle, biology, and medical significance of tabanids. PMID- 10673460 TI - Identifying and misidentifying the brown recluse spider. AB - The brown recluse spider, Loxosceles reclusa, is often implicated as a cause of necrotic skin lesions.[1-3] Diagnoses are most commonly made by clinical appearance and infrequently is a spider seen, captured or identified at the time of the bite.[1, 2, 4-6] The brown recluse lives in a circumscribed area of the U.S. (the south central Midwest) with a few less common recluse species living in the more sparsely-populated southwest U.S.[7] In these areas, where spider populations may be dense, recluse spiders may be a cause of significant morbidity. However, outside the natural range of these recluse species, the conviction that they are the etiological agents behind necrotic lesions of unknown origin is widespread, and most often erroneous. In some states such as California, unsubstantiated reports concerning recluse spider bites have taken on the status of "urban legend" leading to overdiagnosis and, therefore, inappropriate treatment. PMID- 10673461 TI - Clinically relevant dermatology resources and the Internet: An introductory guide for practicing physicians. AB - The number of dermatology sites on the Internet is growing rapidly and the resources they offer are of great benefit to dermatologists and physicians. The mainstay of dermatologic diagnosis is that of visual inspection and gross morphology, therefore, the use of improved imaging techniques, which allow for the rapid collection, electronic transmission, and indexing of images aids in the accurate recognition and interpretation of skin lesions. The graphic nature of the field of dermatology kindly lends itself to the use of the Internet. The enhanced ability to access information,and share expert opinion worldwide will help lessen inappropriate treatment or delayed referral to an appropriate specialist and facilitate the accurate diagnosis of various dermatologic diseases. Concerns about the quality of data are examined and an evaluation scheme proposed. An overview of available resources is presented along with links to additional resources to aid the reader in the identification of additional sites to explore. Sites were selected with the goal of identifying those that provide a broad range of information worthwhile to the dermatologist and general practitioner. Resources including, but not limited to, image atlases, electronic texts, drug databases, case study archives, laboratory testing, and clinical procedures are examined. PMID- 10673462 TI - Terbinafine and lupus erythematosus a chance association? PMID- 10673463 TI - Treatment of atopic dermatitis with zafirlukast. PMID- 10673464 TI - Online publishing and repositories. PubMed Central vs. IASTMP. PMID- 10673465 TI - The impact of COPD on lung health worldwide: epidemiology and incidence. AB - Information on the prevalence of COPD was obtained from vital statistics, health interview surveys, hospital charge records, national publications, and the World Health Organization (WHO). These data indicate that COPD is a common disease with implications for global health. In the United States, morbidity caused by COPD is 4%, making COPD the fourth leading cause of death, exceeded only by heart attacks, cancer, and stroke. Internationally, there is substantial variation in death rates possibly reflecting smoking behavior, type and processing of tobacco, pollution, climate, respiratory management, and genetic factors. The Global Obstructive Lung Disease Initiative, initiated by the National Heart, Lung, and Blood Institute and the WHO, aims to raise awareness of the increasing burden of COPD, decrease morbidity and mortality, promote further study of the condition, and implement programs to prevent COPD. PMID- 10673466 TI - The economic burden of COPD. AB - COPD is one of the leading causes of morbidity and mortality worldwide and imparts a substantial economic burden on individuals and society. Despite the intense interest in COPD among clinicians and researchers, there is a paucity of data on health-care utilization, costs, and social burden in this population. The total economic costs of COPD morbidity and mortality in the United States were estimated at $23.9 billion in 1993. Direct treatments for COPD-related illness accounted for $14.7 billion, and the remaining $9.2 billion were indirect morbidity and premature mortality estimated as lost future earnings. Similar data from another US study suggest that 10% of persons with COPD account for > 70% of all medical care costs. International studies of trends in COPD-related hospitalization indicate that although the average length of stay has decreased since 1972, admissions per 1,000 persons per year for COPD have increased in all age groups > 45 years of age. These trends reflect population aging, smoking patterns, institutional factors, and treatment practices. PMID- 10673467 TI - Mechanisms in COPD: differences from asthma. AB - Although considerable progress has been made in understanding the cellular and molecular mechanisms of asthma, much less attention has been paid to COPD. The inflammatory process in COPD is very different from that in asthma, with different inflammatory cells, mediators, inflammatory effects, and response to therapy. Airway inflammation in asthma, characterized by an eosinophilic inflammation affecting all the airways but not lung parenchyma, is linked to airway hyperresponsiveness. In COPD, there is a predominantly neutrophilic inflammation in the airways. Parenchymal destruction is an important irreversible feature and leads to airflow obstruction through dynamic compression. The eosinophilic inflammation in asthma is markedly suppressed by corticosteroids, but they have no appreciable effect on the inflammation in COPD, consistent with a failure of long-term corticosteroids to alter the progression of COPD. PMID- 10673468 TI - The importance of spirometry in COPD and asthma: effect on approach to management. AB - COPD is characterized by airflow limitation. The diagnosis is suggested by history and physical examination and is confirmed by spirometry (ie, a low FEV(1) level that is unresponsive to bronchodilators). Once diagnosed, there is no widely accepted staging or severity scoring system. COPD presently is graded using a single measurement such as FEV(1), which, unlike the case with asthma, has a limited role in disease management. A more comprehensive staging system is required incorporating, for example, age, arterial blood gases, dyspnea, body mass index, and distance walked, in addition to FEV(1). These criteria should allow for more evidence-based recommendations for management of this condition. Asthma is an inflammatory disease also characterized by airflow limitation. But in contrast with COPD, the airflow limitation is highly reversible either spontaneously or with therapy. Repeated lung function measurements using portable peak flowmeters have resulted in improved outcomes. Therefore, frequent flow determination is recommended in the routine management of asthma. Treatment with anti-inflammatory agents and close monitoring of lung function should help decrease the morbidity and mortality associated with asthma. PMID- 10673469 TI - National and international guidelines for COPD: the need for evidence. AB - The introduction of guidelines for the management of asthma has led to standardization of management and better care of patients with the condition. Many national and international respiratory societies have developed guidelines for COPD. The World Health Organization and the National Heart, Lung, and Blood Institute are jointly developing guidelines that will present evidence-based recommendations for the management of COPD. The guidelines will discuss the definition, epidemiology, natural history, risk factors, pathology, and diagnosis of COPD. There will be guidance on the management of chronic disease and acute exacerbations, education, prevention, and socioeconomics. PMID- 10673470 TI - Recommendations for the management of COPD. AB - Three sets of guidelines for the management of COPD that are widely recognized (from the European Respiratory Society [ERS], American Thoracic Society [ATS], and British Thoracic Society [BTS]) are reviewed and compared. None of the documents uses classic evidence-based documentation, and, in many instances, the recommendations are empiric because of a lack of scientific evidence. Overall, there is strong agreement between the documents. All three guidelines recommend inhaled bronchodilators as first-line therapy. Anticholinergics are noted to be well tolerated, although potential problems with beta(2)-agonists are mentioned. The ERS and BTS suggest that inhaled corticosteroids may be of value in patients documented to be steroid responders, whereas the ATS does not recommend their use at all. All three guidelines support the use of oxygen and pulmonary rehabilitation. There are varying levels of disagreement between the guidelines related to the role of spirometry, stratification of disease severity, and the use of theophylline and systemic corticosteroids. Other differences include the role for nebulizers and metered-dose inhalers, secretion clearance methodologies, and the treatment of acute COPD exacerbations and acute respiratory failure. All three guidelines agree that more research is needed to improve our understanding and management of COPD. PMID- 10673471 TI - The reality of drug use in COPD: the European perspective. AB - COPD guidelines provide advice about the appropriate use of various medications in treating patients with this condition. Comparisons of drug therapy as recommended by these guidelines with what is actually prescribed by both primary care physicians and specialist pulmonologists in a number of European countries can be examined in a variety of ways. Nonadherence to guidelines and differences between countries are caused by a number of factors, including varying degrees of misdiagnosis and different national attitudes to various classes of drugs. PMID- 10673472 TI - Suboptimal medical therapy in COPD: exploring the causes and consequences. AB - Effective outpatient management of COPD requires prescription of and adherence to appropriate therapies. Although practice guidelines for outpatient management of COPD are widely available, evidence suggests that these guidelines are not being implemented widely in clinical practice. Furthermore, several studies have shown that patient compliance with recommended therapy is poor. This paper discusses several reasons why implementation of practice guidelines and adherence with prescribed therapies may be poor. Potential clinical and economic consequences of suboptimal management are reviewed. Although the evidence suggests that improved compliance with guideline-recommended practice will improve symptoms and disease specific quality of life, further work needs to be done to establish the cost effectiveness of chronic therapies for COPD relative to other chronic conditions. Without such data, managed care organizations will be reluctant to allocate scarce resources toward expensive guideline implementation programs for individuals with this condition. PMID- 10673473 TI - How can the implementation of guidelines be improved? AB - Guidelines for a variety of diseases have now been produced. However, implementation of guidelines requires that the medical profession is willing to conform to patterns of diagnostic and treatment behavior set down by others. This may not happen in practice. Early experience in the United Kingdom was gained with the introduction of guidelines for the management of asthma. For a number of years, there have been improvements in practice, but deficiencies still exist. When the introduction of guidelines for the management of COPD was planned, a new approach was taken with a consortium of the British Thoracic Society, pharmaceutical companies, and medical equipment companies being formed to promote their use. Early studies show that COPD care starts from an even lower baseline than asthma; there is poor understanding of objective diagnosis of COPD in both primary and secondary care. PMID- 10673474 TI - Assessment of bronchodilator efficacy in symptomatic COPD: is spirometry useful? AB - Bronchodilator therapy in COPD is deemed successful if it improves ventilatory mechanics to a degree where effective symptom alleviation and increased exercise capacity are achieved. A greater understanding of the pathophysiologic mechanisms of dyspnea and exercise intolerance in COPD has prompted a reevaluation of the manner in which we currently assess therapeutic efficacy. The traditional reliance on an improved postbronchodilator FEV(1) as indicative of a positive clinical response has recognized limitations. To the extent that pharmacologic volume reduction is a desirable therapeutic goal with favorable implications for dyspnea relief and increased exercise tolerance, the potential value of bronchodilator-induced changes in lung volume measurements is currently being studied. It is unlikely, however, given the multifactorial nature of dyspnea and exercise limitation in COPD, that resting spirometric measurements of maximal flows and volumes alone will be sufficiently sensitive to adequately predict a positive clinical response to bronchodilator therapy. Thus, additional direct measurements of exercise dynamic hyperinflation and exercise endurance together with reliable subjective measurements of dyspnea and quality of life are recommended in the setting of a suitable placebo-controlled design. PMID- 10673475 TI - Impact of sleep in COPD. AB - Sleep has well-recognized effects on breathing, including changes in central respiratory control, airways resistance, and muscular contractility, which do not have an adverse effect in healthy individuals but may cause problems in patients with COPD. Sleep-related hypoxemia and hypercapnia are well recognized in COPD and are most pronounced in rapid eye movement sleep. However, sleep studies are usually only indicated in patients with COPD when there is a possibility of sleep apnea or when cor pulmonale and/or polycythemia are not explained by the awake PaO(2) level. Management options for patients with sleep-related respiratory failure include general measures such as optimizing therapy of the underlying condition; physiotherapy and prompt treatment of infective exacerbations; supplemental oxygen; pharmacologic treatments such as bronchodilators, particularly ipratropium bromide, theophylline, and almitrine; and noninvasive positive pressure ventilation. PMID- 10673476 TI - How should health-related quality of life be assessed in patients with COPD? AB - The traditional approach of caring for patients with chronic respiratory disease has been to rely on pulmonary function tests to quantify the severity and to assess response to therapy. However, patients with respiratory conditions seek medical attention because of symptoms, particularly dyspnea, and impaired ability to function, which clearly impact on an individual's health-related quality of life (HRQOL). Accordingly, instruments have been developed to provide a standardized method to measure health status and levels of impairment. One of the major reasons for measuring HRQOL is to detect how much HRQOL has changed in response to therapy (an evaluative instrument). A minimum clinically significant change has been established for some HRQOL instruments in order to indicate the relative value of any measured change and to guide the interpretation as to whether the change is "clinically meaningful." Selected studies using disease specific instruments have demonstrated that beta(2)-agonist, anticholinergic, and theophylline medications can improve HRQOL, as compared with placebo therapy. PMID- 10673477 TI - New approaches to the management of COPD. AB - Airflow limitation in COPD is a result partially of bronchospasm, but it is also caused by a reduction in airway caliber, the number of small airways, airway collapse because of loss of connective tissue support, excess mucus in the airways, and edema of the airway wall. Structural changes also occur because of long-term destruction of interstitial connective tissue, including elastin. Therefore, in addition to the traditional aim of reversing bronchospasm with bronchodilators, disease-modifying approaches are being investigated. The enzyme neutrophil elastase is implicated in the induction of bronchial disease causing structural changes in lungs, impairment of mucociliary clearance, and impairment of host defenses. The precise mechanism pathway of neutrophil elastase is uncertain, but the effects of influencing the pathway in order to slow disease progression are being investigated. Oxidants may also have a role in the development of COPD, with increased levels activating airway cells and cytokine production. PMID- 10673478 TI - The pharmacological properties of tiotropium. AB - Tiotropium is a long-acting anticholinergic drug. Studies with cloned human muscarinic receptors show that tiotropium binds equally well to M(1), M(2), and M(3) receptors. However, it dissociates very slowly from M(1) and M(3) receptors compared with ipratropium, and more rapidly from M(2) receptors. Binding studies with [(3)H]tiotropium in human lung show that it is approximately 10-fold more potent than ipratropium. In vitro, tiotropium has a potent inhibitory effect against cholinergic nerve-induced contraction of airways. It dissociates extremely slowly, compared with the dissociation of atropine and ipratropium. Clinical studies with single doses of inhaled tiotropium confirm that it is a potent and long-lasting bronchodilator. Furthermore, it protects against cholinergic bronchoconstriction for > 24 h. Pharmacokinetic studies show that little of the inhaled drug is absorbed, thus predicting a high margin of safety. PMID- 10673479 TI - The future for tiotropium. AB - In spite of growing interest in and knowledge about the causes and progression of COPD, neither the assessment nor pharmacologic treatment of this condition is currently ideal. Tiotropium is the first new treatment for COPD in many years. Tiotropium is a long-acting anticholinergic agent that has a potential role as a once-daily maintenance treatment, and a picture of its effectiveness is gradually emerging. Spirometry data from clinical studies demonstrate that it is a potent bronchodilator in patients with COPD and it is very well tolerated. Further data on health status and quality of life are awaited. PMID- 10673480 TI - Primary care research: ends and means. PMID- 10673481 TI - Knowledge and attitudes of primary care physicians regarding battered women. Comparison between specialists in family medicine and GPs. AB - BACKGROUND: Domestic violence is a widespread public health problem and an important part of primary care practice. OBJECTIVE: To evaluate the approach of primary care physicians (family physicians and GPs) to the care of battered women. METHODS: A self-report questionnaire containing items about experience, knowledge and attitudes regarding the care of battered women was mailed to a random sample of 300 primary care physicians employed by the two major health management organizations in Israel. The population included family physicians, who have 4 years of residency training in primary care, and GPs, who do not undergo specialization after completing their medical studies. RESULTS: A total of 236 physicians (130 family physicians and 106 GPs) responded. In general, the physicians had had very little exposure to the problem and estimated its prevalence in the community as less than half that indicated in the medical literature. Compared with the GPs, however, the family physicians reported more exposure to the subject (P < 0.001) and had better knowledge of its prevalence and risk factors (P < 0.001). They also showed a greater tendency to view the problem as universal (P < 0.05) and as part of their professional responsibilities. However, both groups tended not to include the care of battered women with no physical injury within their professional duties. CONCLUSIONS: Physicians should be made more aware of the problem of battered women within the context of their routine professional practice and of the importance of keeping abreast of the subject. Educators should place more emphasis on imparting knowledge and skills in the management of battered women, especially for GPs. PMID- 10673482 TI - The work by the developing primary care team in China: a survey in two cities. AB - BACKGROUND: China is in the process of converting its existing primary care resources into general practice. The infrastructure is different from that of many other countries. OBJECTIVES: We surveyed patients' reasons for encounter (RFE) and the health providers' diagnoses in the general practice clinics of two large northern cities in order to assess the nature of the work of these practices. METHOD: Practices whose staff had a short course of training in the theory and practice of the International Classification of Primary Care (ICPC) were recruited to document the RFE and diagnoses of patient encounters in two separate winter weeks. RESULTS: The practices dealt mainly with chronic illness in older patients. Hypertension-related problems were the most frequent diagnoses, followed by upper respiratory tract infection. Patients also consulted very frequently for dizziness. Overall, there was good agreement between RFE and diagnosis in some organ systems. CONCLUSION: In their present form, the Chinese practices surveyed were delivering the full range of general practice care to a self-selected age group of patients. The ICPC was very useful for monitoring the work of general practice from the perspective of both the patients and the providers. PMID- 10673483 TI - Continuity of care: towards a definition based on experiences of practising GPs. AB - BACKGROUND: The traditional concept of continuity of care, i.e. care from the cradle to the grave, is no longer sustainable in modern society. OBJECTIVE: The aim of this study was to propose a definition of 'continuity of care' based on the experiences of a group of practising Australian GPs. METHOD: Five focus group discussions were conducted to explore the understanding and practice of continuity of care, the individual's measurement of having achieved continuity of care in his/her practice and the advantages/disadvantages of providing continuity of care. Results and conclusions. The experiences of this group of GPs points towards three essential aspects to help with a definition of continuity of care. Firstly it requires a stable care environment, secondly good communication to build a responsible doctor-patient relationship and thirdly the goal of achieving an improvement of the patient's overall health. PMID- 10673484 TI - Patient perception of quality following a visit to a doctor in a primary care unit. AB - BACKGROUND: Assessment of the quality of primary care services may be enhanced by including patient perceptions as well as professional judgment of quality. There is a need for reliable and valid instruments to measure these perceptions. OBJECTIVES: (i) To present a scale for measuring patient perception of quality of care following a visit to a doctor; and (ii) to analyse the responses given by patients recruited in primary care units in the Montreal region. The scale is composed of 22 items regrouped into three sub-scales referring to the patient physician relationship (five items); the technical aspects of care (12 items); and the outcomes of the visit (five items). Distinctive features of the scale are that it focuses on patients' opinions about quality rather than on satisfaction, and that it includes items related to outcomes of the visit. METHODS: A survey was conducted on 473 patients who visited a physician in 11 primary care units in the Montreal region. Randomly selected patients received mailed questionnaires 5 7 days following their visit. Various statistical procedures were used to assess the reliability and the validity of the global scale and the sub-scales, and to analyse patients' patterns of response. RESULTS: The analysis of the psychometric properties of the global scale and the three sub-scales provides favourable evidence concerning their reliability and validity. The results of the factor analysis, the inter-item correlations and the Cronbach's alpha coefficients all support the distinction made between the interpersonal processes, the technical processes and the outcomes, and, at the same time, confirm the complex nature of the notion of perceived quality. The analysis of patients' responses allows the identification of items associated with global perception about quality of care. This global perception results from patients' perception of the physician's professional and interpersonal skills as well as from the outcomes of care. CONCLUSION: The scale can be used by physicians or primary health care units and has a wide range of applications. PMID- 10673485 TI - Which literature retrieval method is most effective for GPs? AB - BACKGROUND: Evidence-based medicine requires new skills of physicians, including literature searching. OBJECTIVE: To determine which literature retrieving method is most effective for GPs: the printed Index Medicus; Medline through Grateful Med; or Medline on CD-ROM. METHODS: The design was a randomized comparative study. In a continuing medical education course, three groups of health care professionals (87 GPs and 16 other health care professionals) used one of the literature retrieval methods to retrieve citations on four search topics related to general practice. For the analysis in pairs, we used the search results of the 75 participants who completed all four assignments. As outcome measures, we used precision, recall and an overall search quality score; we also had a post-course questionnaire on personal characteristics, experience with computers, handling medical literature and satisfaction with course instruction and search results. RESULTS: The recall and overall search quality scores in the Index Medicus groups (n = 32) were higher (P = <0.001) than those in the CD-ROM groups (n = 31). In addition, the search quality scores in the Grateful Med groups (n = 12) were higher (P < 0.003) than those in the CD-ROM groups. There were no differences in precision. CONCLUSION: In the period 1994-1997, the printed Index Medicus was the most effective literature retrieval method for GPs. For inexperienced GPs, there is a need for training in electronic literature retrieval methods. PMID- 10673486 TI - A study of factors associated with cost and variation in prescribing among GPs. AB - BACKGROUND: Inappropriate prescribing has the potential to harm both the individual and society. Previous research has identified doctor or demographic characteristics that influence prescribing variation but which were not amenable to change. OBJECTIVES: To identify modifiable factors associated with GP prescribing variance and cost. METHOD: Qualitative research methods were used in semi-structured taped interviews with 17 GPs in Avon, South West NHS Region, UK. RESULTS: GPs considered themselves cautious and conservative prescribers. Prescribing decisions often were justified by the prescriber, despite conflicting clinical or cost arguments. A personally developed drug formulary was used to reduce dilemmas potentially associated with prescribing uncertainty. Willingness to reflect upon, and measure, prescribing habits against set professional standards varied considerably. The absence of monitoring mechanisms of prescribing decisions, coupled with under utilization of the community pharmacist, resulted in uncertain prescribing outcomes. Some GPs found it difficult to keep up to date professionally due to perceived time constraints. Excessive patient demand was considered to influence their prescribing, but GPs stated that they were not unduly influenced by the drug representative. CONCLUSIONS: Prescribing makes a considerable impact on health and budgets and yet remains a contentious issue. Improved partnerships between patient, doctor and pharmacist must be established. Better prescribing decision monitoring and support through policy development and educational intervention is needed to reduce prescribing uncertainty. Newly established Primary Care Groups may need to reflect upon the difficulties facing prescribers, particularly when prescribing within cash-limited budgets, to avoid discord between prescribing behaviour and local policy development. PMID- 10673487 TI - Patient adherence to family practitioners' recommendations for breast cancer screening: a historical cohort study. AB - BACKGROUND: Breast cancer is the most prevalent malignancy among women in Israel, and routine screening is recommended for early detection. In 1997, a health management organization primary care centre in rural Israel established a 1-year programme wherein family physicians were encouraged to remind their patients to undergo breast cancer examinations. This study evaluates the impact of the physicians' intervention on patient compliance. METHODS: Family practitioners from two practices were requested to discuss the importance of early breast cancer detection with all eligible patients who visited the clinic for any reason and to assist them in scheduling an appointment for screening. The files of the patients who received the recommendation were stamped accordingly. On completion of the programme, the physicians' files were audited, and the potential candidates for breast cancer screening were divided into two groups: those who had received the intervention (n = 251) and those who had not (n = 187); results were also compared with those of a third group of patients who had gone for an examination on their own initiative (n = 100) prior to the study (i.e. did not require intervention). A random sample of half the patients also completed an ad hoc questionnaire covering sociodemographic variables and the impact of the doctors' intervention on their behaviour. RESULTS: Patients in the intervention group showed a significantly greater change in behaviour regarding breast cancer screening than the controls (32% versus 13%, P = 0.001). This change was manifested particularly in the group of women aged 50-74 years who received the recommendation for mammography to be performed (according to the guidelines). CONCLUSION: Although this is a study in only two practices, the results suggest that primary care physicians can significantly alter the behaviour of their patients regarding regular breast examinations. The use of a special reminder can also help the individual doctor to ensure that each patient has been properly instructed. PMID- 10673488 TI - Influence of postal distribution of the Royal College of Radiologists' guidelines, together with feedback on radiological referral rates, on X-ray referrals from general practice: a randomized controlled trial. AB - BACKGROUND: The Royal College of Radiologists (RCR) have produced regularly updated guidelines on radiological referrals since 1990. A small study in 1992 showed postal distribution of guidelines reduced general practitioners' referrals over the subsequent 9 weeks. However there have been no randomized trials of the longer term effects of radiological guidelines and feedback on referral rates on X-ray requests from primary care. OBJECTIVES: To see if the introduction of radiological guidelines into general practices together with feedback on referral rates reduces the number of GP radiological requests over one year; and to explore GPs'attitudes to the guidelines. METHODS: Sixty-nine practices referring patients to St George's Healthcare Trust were randomly allocated to intervention or control groups. In February 1995 a GP version of the RCR guidelines was sent to each GP in the 33 practices in the intervention group. After 9 months intervention, practices were sent revised guidelines with individual feedback on the number of examinations requested in the past 6 months. The total number of requests per practice was compared for the year before and the year after the introduction of the guidelines. Control practices were sent the guidelines at the end of the study. All doctors were sent a questionnaire about the guidelines. RESULTS: A total of 43 778 radiological requests were made during the two years 1994-1996. In practices receiving the guidelines there was a 20% reduction in requests for spinal examinations compared with control practices (P < 0.05). This corresponded to the effect reported by GPs. There was also a 10% difference between the groups in the total number of requests made, but due to wide interpractice variation in referral rates this failed to reach statistical significance. CONCLUSIONS: Introduction of radiological guidelines together with feedback on referral rates was effective in reducing the number of requests for spinal examinations over one year. Wider use of GP-orientated guidelines with regular updating and feedback might save costs and reduce unnecessary irradiation of patients. PMID- 10673489 TI - GPs' employment of locum doctors and satisfaction with their service. AB - BACKGROUND: Locum doctors provide cover during normal working hours for GPs absent due to holidays, sickness, maternity leave or for educational purposes. However, there is little information on the extent of the use of locums or of GPs' perception of their services. OBJECTIVES: To examine the level of use of locum doctors by GPs, the ease of recruitment and satisfaction with their services. METHODS: A postal survey of all general practices in one of the six health regions in England was carried out. Logistic regression analysis was used to examine the independent effects on locum use of practice size and type of area, source of recruitment and GPs' satisfaction with their services. RESULTS: A total of 935 (80.6%) general practices responded. Locum GPs were employed by 81.7% of practices in the previous 12 months. Two-thirds of practices reported problems obtaining locum cover, especially at short notice and for holiday periods. One-fifth of practices employing a locum in the previous 12 months were dissatisfied with the locum. CONCLUSIONS: There are high demands for, but a considerable shortage of, locum doctors in general practice. Educational and other initiatives for GPs may contribute to increased demands for locum cover. Difficulties in recruitment may be reduced by measures to improve the conditions of employment for doctors working as locums on a longer term basis. New codes of practice for employing locums may increase satisfaction with locum services. PMID- 10673490 TI - Condom promotion in women attending inner city general practices for cervical smears: a randomized controlled trial. AB - BACKGROUND: Although condom promotion schemes have been widely piloted in UK general practices, there have been no rigorous evaluations of their effectiveness. OBJECTIVES: To see if a practice-based sexual health education intervention increases the number of women having smears who are given condoms and advice on avoiding STDs. To see if this low cost intervention affects subsequent condom use. METHOD: We conducted a cluster randomized trial of condom promotion in 1382 women aged <35 years attending 28 South London general practices for cervical smear tests. RESULTS: More women in intervention than control practices reported receiving advice on avoiding sexually transmitted infections (27% versus 10%) and being given condoms (28% versus 1%, P < 0.05). However, there was no difference in subsequent condom use, even in the 22% of women reporting >/=2 sexual partners in the previous year. CONCLUSIONS: To provide evidence of effectiveness, future interventions may need to be more intensive or focus on higher risk groups. PMID- 10673491 TI - A pilot study of cardiovascular risk assessment in Afro-Caribbean patients attending an inner city general practice. AB - BACKGROUND: Afro-Caribbean ethnic minorities are at high risk of stroke and the sequelae of hypertension. OBJECTIVE: To investigate cardiovascular risk factors and Dundee risk rank in Afro-Caribbeans attending one inner city general practice and to find which methods of health promotion patients preferred. METHODS: We assessed cardiovascular risk including systolic and diastolic blood pressure in 98 patients of Afro-Caribbean origin. RESULTS: Fifty per cent of the patients had at least two risk factors for cardiovascular disease. Focus groups suggested that the barriers to effective health promotion included lack of risk awareness, cultural and lifestyle influences, time restrictions and language difficulties. CONCLUSIONS: The small pilot study highlights both the need for and some of the problems of GP-based cardiovascular health promotion in Afro-Caribbeans. PMID- 10673492 TI - Criteria for selecting and using non-steroidal anti-inflammatory drugs in primary health care. AB - BACKGROUND: Numerous non-steroidal anti-inflammatory drugs (NSAIDs) are commercially available in Spain. This makes proper selection and use of these drugs difficult for GPs. OBJECTIVE: To find out the criteria which GPs use to choose among the drugs in this group and to evaluate other criteria related to the clinical management of NSAIDs. METHOD: A survey questionnaire was distributed among all the GPs of the reformed network in the health area of Tarragona (Spain). RESULTS: The doctors questioned chose a limited number of NSAIDs. Major preferences were diclofenac and piroxicam. The main criteria for selection were: efficacy and safety of the drug; previous and/or current gastrointestinal pathology of the patient; the difference between acute and chronic use; and the type of inflammatory process. There were a large number of combinations of routes of administration. The histamine H2 antagonists were the prophylactic drugs which were used most for gastropathy. CONCLUSION: Sometimes the main criteria for choosing between the drugs in this study do not reflect the drugs that are used in clinical practice. Some of the criteria used by GPs for selecting and using NSAIDs should be reviewed. PMID- 10673493 TI - Costing interventions in primary care. AB - Against a background of increasing demands on limited resources, studies that relate benefits of health interventions to the resources they consume will be an important part of any decision-making process in primary care, and an accurate assessment of costs will be an important part of any economic evaluation. Although there is no such thing as a gold standard cost estimate, there are a number of basic costing concepts that underlie any costing study. How costs are derived and combined will depend on the assumptions that have been made in their derivation. It is important to be clear what assumptions have been made and why in order to maintain consistency across comparative studies and prevent inappropriate conclusions being drawn. This paper outlines some costing concepts and principles to enable primary care practitioners and researchers to have a basic understanding of costing exercises and their pitfalls. PMID- 10673494 TI - The 'doctor' or the 'girl from the University'? Considering the influence of professional roles on qualitative interviewing. AB - BACKGROUND: Qualitative research methods are now recognized as valuable tools for primary care. With the increasing emphasis on evidence-based medicine and critical appraisal of published work, it is important that qualitative researchers are transparent about their methods and discuss the impact of the research process on their data. OBJECTIVES: To consider the impact of the professional background of researchers on in-depth interviewing in primary care. METHODS: We compare interactions between the interviewer and respondents in two qualitative interview studies of heart disease. Both samples consisted of 60 middle-aged men and women from a range of social backgrounds living in the West of Scotland. One study was conducted by a GP and the other by a sociologist. RESULTS: Some interview interactions were common to both researchers; for example, interviews were often regarded by respondents as therapeutic. However, some interactions seemed to be related to the researcher's professional background. The GP's perceived higher status led to obscuring of her personal characteristics. The sociologist was often perceived as a 'young woman' rather than defined by her professional role. Thus respondents' perceptions of the interviewer influenced the interview interactions. CONCLUSIONS: Appraising qualitative research depends on the transparency with which the research process is described. Awareness of professional background is particularly important for university departments of primary care (which often include doctors, nurses and social scientists) and should be considered carefully in designing, carrying out and disseminating the results of qualitative studies. PMID- 10673495 TI - Acquiring qualitative skills for primary care research. Review and reflections on a three-stage workshop. Part 1: using interviews to generate data. Members of WoReN. Wolds Primary Care Research Network. AB - This paper reflects on one Primary Care Research Network's (WoReN's) experience of running a workshop on generating interview data, provided as the first of a three-part workshop concerned with acquiring qualitative interviewing skills. It discusses the aims and limitations of the short workshop format in meeting the needs of practitioners embarking on qualitative research, drawing upon and reviewing the relevant research methods literature, and makes suggestions with regard to designing and running research methods workshops within primary care. PMID- 10673496 TI - Acquiring qualitative skills for primary care research. Review and reflections on a three-stage workshop. Part 2: analysing interview data. Members of WoReN. Primary Care Research Network. AB - This paper reflects on one Primary Care Research Network's (WoReN's) experience of running a workshop on analysing qualitative interview data, provided as the second of a three-part workshop concerned with acquiring qualitative interviewing skills. It discusses the aims and limitations of the short workshop format in meeting the needs of practitioners embarking on the process of analysing qualitative data, drawing upon and reviewing the relevant research methods literature. Particular attention is paid to the role of qualitative data analysis computer packages and the debate on 'grounded theory'. We conclude by making suggestions with regard to designing and running data analysis workshops within primary care. PMID- 10673497 TI - Selections from current literature. Psychoneuroimmunology: validation of the biopsychosocial model. PMID- 10673498 TI - Light: an indicator of time and place. PMID- 10673499 TI - Gene dose-dependent control of hematopoiesis and hematologic tumor suppression by CBP. AB - Mice with monoallelic inactivation of the CBP gene develop highly penetrant, multilineage defects in hematopoietic differentiation and, with advancing age, an increased incidence of hematologic malignancies. The latter are characterized, at least in some cases, by loss of heterozygosity (LOH) at the CBP locus. No such pathology was observed in wild-type or p300 heterozygous null mice of the same age and genetic background. Thus, a full complement of CBP, but not p300, is required for normal hematopoietic differentiation. These results also provide the first experimental evidence for the hypothesis that CBP has tumor-suppressing activity. PMID- 10673500 TI - Chk2/hCds1 functions as a DNA damage checkpoint in G(1) by stabilizing p53. AB - Chk2/hcds1, the human homolog of the Saccharomyces cerevisiae RAD53/SPK1 and Schizosaccharomyces pombe cds1 DNA damage checkpoint genes, encodes a protein kinase that is post-translationally modified after DNA damage. Like its yeast homologs, the Chk2/hCds1 protein phosphorylates Cdc25C in vitro, suggesting that it arrests cells in G(2) in response to DNA damage. We expressed Chk2/hCds1 in human cells and analyzed their cell cycle profile. Wild-type, but not catalytically inactive, Chk2/hCds1 led to G(1) arrest after DNA damage. The arrest was inhibited by cotransfection of a dominant-negative p53 mutant, indicating that Chk2/hCds1 acted upstream of p53. In vitro, Chk2/hCds1 phosphorylated p53 on Ser-20 and dissociated preformed complexes of p53 with Mdm2, a protein that targets p53 for degradation. In vivo, ectopic expression of wild-type Chk2/hCds1 led to increased p53 stabilization after DNA damage, whereas expression of a dominant-negative Chk2/hCds1 mutant abrogated both phosphorylation of p53 on Ser-20 and p53 stabilization. Thus, in response to DNA damage, Chk2/hCds1 stabilizes the p53 tumor suppressor protein leading to cell cycle arrest in G(1). PMID- 10673501 TI - The human homologs of checkpoint kinases Chk1 and Cds1 (Chk2) phosphorylate p53 at multiple DNA damage-inducible sites. AB - Upon DNA damage, the amino terminus of p53 is phosphorylated at a number of serine residues including S20, a site that is particularly important in regulating stability and function of the protein. Because no known kinase has been identified that can modify this site, HeLa nuclear extracts were fractionated and S20 phosphorylation was followed. We discovered that a S20 kinase activity copurifies with the human homolog of the Schizosaccharomyces pombe checkpoint kinase, Chk1 (hCHK1). We confirmed that recombinant hCHK1, but not a kinase-defective version of hCHK1, can phosphorylate p53 in vitro at S20. Additional inducible amino- and carboxy-terminal sites in p53 are also phosphorylated by hCHK1, indicating that this is an unusually versatile protein kinase. It is interesting that hCHK1 strongly prefers tetrameric to monomeric p53 in vitro, consistent with our observation that phosphorylation of amino-terminal sites in vivo requires that p53 be oligomeric. Regulation of the levels and activity of hCHK1 in transfected cells is directly correlated with the levels of p53; expression of either a kinase-defective hCHK1 or antisense hCHK1 leads to reduced levels of cotransfected p53, whereas overexpression of wild-type hCHK1 or the kinase domain of hCHK1 results in increased levels of expressed p53 protein. The human homolog of the second S. pombe checkpoint kinase, Cds1 (CHK2/hCds1), phosphorylates tetrameric p53 but not monomeric p53 in vitro at sites similar to those phosphorylated by hCHK1 kinase, suggesting that both checkpoint kinases can play roles in regulating p53 after DNA damage. PMID- 10673502 TI - c-Kit triggers dual phosphorylations, which couple activation and degradation of the essential melanocyte factor Mi. AB - Microphthalmia (Mi) is a bHLHZip transcription factor that is essential for melanocyte development and postnatal function. It is thought to regulate both differentiated features of melanocytes such as pigmentation as well as proliferation/survival, based on phenotypes of mutant mouse alleles. Mi activity is controlled by at least two signaling pathways. Melanocyte-stimulating hormone (MSH) promotes transcription of the Mi gene through cAMP elevation, resulting in sustained Mi up-regulation over many hours. c-Kit signaling up-regulates Mi function through MAP kinase phosphorylation of Mi, thereby recruiting the p300 transcriptional coactivator. The current study reveals that c-Kit signaling triggers two phosphorylation events on Mi, which up-regulate transactivation potential yet simultaneously target Mi for ubiquitin-dependent proteolysis. The specific activation/degradation signals derive from MAPK/ERK targeting of serine 73, whereas serine 409 serves as a substrate for p90 Rsk-1. An unphosphorylatable double mutant at these two residues is at once profoundly stable and transcriptionally inert. These c-Kit-induced phosphorylations couple transactivation to proteasome-mediated degradation. c-Kit signaling thus triggers short-lived Mi activation and net Mi degradation, in contrast to the profoundly increased Mi expression after MSH signaling, potentially explaining the functional diversity of this transcription factor in regulating proliferation, survival, and differentiation in melanocytes. PMID- 10673503 TI - Transient depletion of xDnmt1 leads to premature gene activation in Xenopus embryos. AB - In Xenopus laevis zygotic transcription begins at the midblastula transition (MBT). Prior to this the genome is organized into chromatin that facilitates rapid cycles of DNA replication but not transcription. Here we demonstrate that DNA methylation contributes to the overall transcriptional silencing before MBT. Transient depletion of the maternal DNA methyltransferase (xDnmt1) by anti sense RNA during cleavage stages is associated with a decrease in the genomic 5-methyl cytosine content and leads to the activation of zygotic transcription approximately two cell cycles earlier than normal. Hypomethylation allows the early expression of mesodermal marker genes such as Xbra, Cerberus, and Otx2, which are subsequently down-regulated during gastrulation of the xDnmt1-depleted embryos. The temporal switch in gene expression may account for the appearance of body plan defects that we observe. Loss of xDnmt1 can be rescued by the coinjection of mouse or human Dnmt1 protein. These results demonstrate that DNA methylation has a role in the regulation of immediately early genes in Xenopus at MBT. PMID- 10673504 TI - Absence of Wee1 ensures the meiotic cell cycle in Xenopus oocytes. AB - Meiotic cells undergo two successive divisions without an intervening S phase. However, the mechanism of S-phase omission between the two meiotic divisions is largely unknown. Here we show that Wee1, a universal mitotic inhibitor, is absent in immature (but not mature) Xenopus oocytes, being down-regulated specifically during oogenesis; this down-regulation is most likely due to a translational repression. Even the modest ectopic expression of Wee1 in immature (meiosis I) oocytes can induce interphase nucleus reformation and DNA replication just after meiosis I. Thus, the presence of Wee1 during meiosis I converts the meiotic cell cycle into a mitotic-like cell cycle having S phase. In contrast, Myt1, a Wee1 related kinase, is present and directly involved in G(2) arrest of immature oocytes, but its ectopic expression has little effect on the meiotic cell cycle. These results strongly indicate that the absence of Wee1 in meiosis I ensures the meiotic cell cycle in Xenopus oocytes. Based on these results and the data published previously in other organisms, we suggest that absence of Wee1 may be a well-conserved mechanism for omitting interphase or S phase between the two meiotic divisions. PMID- 10673505 TI - Activation mutants in yeast RNA polymerase II subunit RPB3 provide evidence for a structurally conserved surface required for activation in eukaryotes and bacteria. AB - We have identified a mutant in RPB3, the third-largest subunit of yeast RNA polymerase II, that is defective in activator-dependent transcription, but not defective in activator-independent, basal transcription. The mutant contains two amino-acid substitutions, C92R and A159G, that are both required for pronounced defects in activator-dependent transcription. Synthetic enhancement of phenotypes of C92R and A159G, and of several other pairs of substitutions, is consistent with a functional relationship between residues 92-95 and 159-161. Homology modeling of RPB3 on the basis of the crystallographic structure of alphaNTD indicates that residues 92-95 and 159-162 are likely to be adjacent within the structure of RPB3. In addition, homology modeling indicates that the location of residues 159-162 within RPB3 corresponds to the location of an activation target within alphaNTD (the target of activating region 2 of catabolite activator protein, an activation target involved in a protein-protein interaction that facilitates isomerization of the RNA polymerase promoter closed complex to the RNA polymerase promoter open complex). The apparent finding of a conserved surface required for activation in eukaryotes and bacteria raises the possibility of conserved mechanisms of activation in eukaryotes and bacteria. PMID- 10673506 TI - Nucleotide excision repair of DNA with recombinant human proteins: definition of the minimal set of factors, active forms of TFIIH, and modulation by CAK. AB - During human nucleotide excision repair, damage is recognized, two incisions are made flanking a DNA lesion, and residues are replaced by repair synthesis. A set of proteins required for repair of most lesions is RPA, XPA, TFIIH, XPC-hHR23B, XPG, and ERCC1-XPF, but additional components have not been excluded. The most complex and difficult to analyze factor is TFIIH, which has a 6-subunit core (XPB, XPD, p44, p34, p52, p62) and a 3-subunit kinase (CAK). TFIIH has roles both in basal transcription initiation and in DNA repair, and several inherited human disorders are associated with mutations in TFIIH subunits. To identify the forms of TFIIH that can function in repair, recombinant XPA, RPA, XPC-hHR23B, XPG, and ERCC1-XPF were combined with TFIIH fractions purified from HeLa cells. Repair activity coeluted with the peak of TFIIH and with transcription activity. TFIIH from cells with XPB or XPD mutations was defective in supporting repair, whereas TFIIH from spinal muscular atrophy cells with a deletion of one p44 gene was active. Recombinant TFIIH also functioned in repair, both a 6- and a 9-subunit form containing CAK. The CAK kinase inhibitor H-8 improved repair efficiency, indicating that CAK can negatively regulate NER by phosphorylation. The 15 recombinant polypeptides define the minimal set of proteins required for dual incision of DNA containing a cisplatin adduct. Complete repair was achieved by including highly purified human DNA polymerase delta or epsilon, PCNA, RFC, and DNA ligase I in reaction mixtures, reconstituting adduct repair for the first time with recombinant incision factors and human replication proteins. PMID- 10673507 TI - RecA protein-dependent R-loop formation in vitro. AB - The RecA protein of Escherichia coli, which has crucial roles in homologous recombination, DNA damage repair, induction of the SOS response, and SOS mutagenesis, was found to catalyze assimilation of complementary RNA into a homologous region of a DNA duplex (R-loop). The reaction strictly requires a region of mismatch in the duplex, which may serve as a nucleation site for RecA protein polymerization. The optimum conditions for the assimilation reaction resemble those for the previously studied RecA protein-catalyzed homologous pairing and strand exchange reaction between two DNA molecules. Our finding lends strong support to the proposal that RecA protein-catalyzed assimilation of a transcript into duplex DNA results in formation of an R-loop at certain regions of the chromosome and that, when stabilized, the R-loop can serve as an origin of chromosome replication. PMID- 10673508 TI - Phosphorylation represses Ets-1 DNA binding by reinforcing autoinhibition. AB - Phosphorylation of transcription factors is a key link between cell signaling and the control of gene expression. Here we report that phosphorylation regulates DNA binding of the Ets-1 transcription factor by reinforcing an autoinhibitory mechanism. Quantitative DNA-binding assays show that calcium-dependent phosphorylation inhibits Ets-1 DNA binding 50-fold. The four serines that mediate this inhibitory effect are distant from the DNA-binding domain but near structural elements required for autoinhibition. Mutational analyses demonstrate that an intact inhibitory module is required for phosphorylation-dependent regulation. Partial proteolysis studies indicate that phosphorylation stabilizes an inhibitory conformation. These findings provide a structural mechanism for phosphorylation-dependent inhibition of Ets-1 DNA binding and demonstrate a new function for inhibitory modules as structural mediators of negative signaling events. PMID- 10673510 TI - Managed care and oncology: the quality debate. PMID- 10673509 TI - Repression by suppressor of hairless and activation by Notch are required to define a single row of single-minded expressing cells in the Drosophila embryo. AB - Notch signal transduction appears to involve the ligand-induced intracellular processing of Notch, and the formation of a processed Notch-Suppressor of Hairless complex that binds DNA and activates the transcription of Notch target genes. This suggests that loss of either Notch or Su(H) activities should lead to similar cell fate changes. However, previous data indicate that, in the Drosophila blastoderm embryo, mesectoderm specification requires Notch but not Su(H) activity. The determination of the mesectodermal fate is specified by Single-minded (Sim), a transcription factor expressed in a single row of cells abutting the mesoderm. The molecular mechanisms by which the dorsoventral gradient of nuclear Dorsal establishes the single-cell wide territory of sim expression are not fully understood. We have found that Notch activity is required for sim expression in cellularizing embryos. In contrast, at this stage, Su(H) has a dual function. Su(H) activity was required to up-regulate sim expression in the mesectoderm, and to prevent the ectopic expression of sim dorsally in the neuroectoderm. We have shown that repression of sim transcription by Su(H) is direct and independent of Notch activity. Conversely, activation of sim transcription by Notch requires the Su(H)-binding sites. Thus, Notch signalling appears to relieve the repression exerted by Su(H) and to up-regulate sim transcription in the mesectoderm. We propose a model in which repression by Su(H) and derepression by Notch are essential to allow for the definition of a single row of mesectodermal cells in the blastoderm embryo. PMID- 10673511 TI - Phase II trial of the antiangiogenic agent thalidomide in patients with recurrent high-grade gliomas. AB - PURPOSE: Little progress has been made in the treatment of adult high-grade gliomas over the last two decades, thus necessitating a search for novel therapeutic strategies. Malignant gliomas are vascular or angiogenic tumors, which leads to the supposition that angiogenesis inhibition may represent a potentially promising strategy in the treatment of these tumors. We present the results of a phase II trial of thalidomide, a putative inhibitor of angiogenesis, in the treatment of adults with previously irradiated, recurrent high-grade gliomas. PATIENTS AND METHODS: Patients with a histologic diagnosis of anaplastic mixed glioma, anaplastic astrocytoma, or glioblastoma multiforme who had radiographic demonstration of tumor progression after standard external-beam radiotherapy with or without chemotherapy were eligible. Patients were initially treated with thalidomide 800 mg/d with increases in dose by 200 mg/d every 2 weeks until a final daily dose of 1,200 mg was achieved. Patients were evaluated every 8 weeks for response by both clinical and radiographic criteria. RESULTS: A total of 39 patients were accrued, with 36 patients being assessable for both toxicity and response. Thalidomide was well tolerated, with constipation and sedation being the major toxicities. One patient developed a grade 2 peripheral neuropathy after treatment with thalidomide for nearly a year. There were two objective radiographic partial responses (6%), two minor responses (6%), and 12 patients with stable disease (33%). Eight patients were alive more than 1 year after starting thalidomide, although almost all with tumor progression. Changes in serum levels of basic fibroblastic growth factor (bFGF) were correlated with time to tumor progression and overall survival. CONCLUSION: Thalidomide is a generally well-tolerated drug that may have antitumor activity in a minority of patients with recurrent high-grade gliomas. Future studies will better define the usefulness of thalidomide in newly diagnosed patients with malignant gliomas and in combination with radiotherapy and chemotherapy. Additionally, studies will be needed to confirm the potential utility of changes in serum bFGF as a marker of antiangiogenic activity and/or glioma growth. PMID- 10673512 TI - Results of a randomized study of IM862 nasal solution in the treatment of AIDS related Kaposi's sarcoma. AB - PURPOSE: Although advances have been made in the treatment of AIDS-related Kaposi's sarcoma (AIDS-KS) with systemic chemotherapy, less toxic therapies are needed. IM862 is a naturally occurring peptide with antiangiogenic properties and was thus studied in patients with AIDS-KS. PATIENTS AND METHODS: IM862 was given as intranasal drops at a dose of 5 mg. Patients were randomized to two dosing schedules given in repeated cycles until disease progression or unacceptable toxicity: 5 days of therapy followed by 5 days off (n = 18) and every other day dosing (n = 26). RESULTS: Forty-two male patients and two female patients with a median age of 38 years (range, 22 to 53 years) were accrued. Twenty-one patients (47%) had more than 50 mucocutaneous lesions, 14 (32%) had lymphedema, and none had visceral involvement. Thirty-three patients (75%) had received prior systemic chemotherapy. Twenty-four patients (55%) had CD4(+) lymphocyte count /= 50 mg/m(2) completed 12 weeks of therapy, and two partial responses were observed. CONCLUSION: C225 has dose dependent pharmacokinetics, and doses that achieve saturation of systemic clearance are well tolerated. C225 given in combination with cisplatin has biologic activity at pharmacologically relevant doses. PMID- 10673535 TI - Phase I and pharmacologic study of oral fluorouracil on a chronic daily schedule in combination with the dihydropyrimidine dehydrogenase inactivator eniluracil. AB - PURPOSE: To determine the maximum-tolerated dose (MTD), toxicities, and pharmacokinetics of oral fluorouracil (5-FU) administered twice daily in combination with oral eniluracil, an inactivator of dihydropyrimidine dehydrogenase, administered for 28 days every 35 days. PATIENTS AND METHODS: Oral 5-FU 1.35 mg/m(2) twice daily was administered with oral eniluracil 10 mg daily for 14 to 28 days, followed by a 1-week rest period. Eniluracil was started 1 day before 5-FU. Patients then received escalated doses of oral 5-FU 1. 35 to 1.8 mg/m(2) twice daily with an increased dose of eniluracil 10 mg twice daily for 28 days. A reduced dose of 5-FU 1.0 mg/m(2) with eniluracil 20 mg twice daily was evaluated. RESULTS: Thirty-six patients with solid malignancies were enrolled onto the study. Diarrhea was the principal dose-limiting toxicity of oral 5-FU and eniluracil given on this chronic schedule. The recommended phase II dose is 5 FU 1.0 mg/m(2) twice daily with eniluracil 20 mg twice daily. Mean (SD) values for terminal half-life, apparent volume of distribution, and systemic clearance of 4.5 hours (0.83 hours), 19 L/m(2) (3.0 L/m(2)), and 51 mL/min/m(2) (13 mL/min/m(2)), respectively. An average of 77% of 5-FU was excreted unchanged in urine after 28 days of treatment. The mean (range) 5-FU C(SS,min) values achieved at the 1.0 mg/m(2) dose level were 22 ng/mL (8 to 38 ng/mL). CONCLUSION: Chronic oral administration of 5-FU with oral eniluracil is tolerable and produces 5-FU steady-state concentrations similar to those achieved with protracted intravenous administration of 5-FU on clinically relevant dose schedules. Eniluracil provides an attractive means of administering 5-FU on protracted schedules. PMID- 10673537 TI - Sector resection for stage I breast cancer. PMID- 10673536 TI - Phase I and pharmacokinetic study of farnesyl protein transferase inhibitor R115777 in advanced cancer. AB - PURPOSE: To determine the maximum-tolerated dose, toxicities, and pharmacokinetic profile of the farnesyl protein transferase inhibitor R115777 when administered orally bid for 5 days every 2 weeks. PATIENTS AND METHODS: Twenty-seven patients with a median age of 58 years received 85 cycles of R115777 using an intrapatient and interpatient dose escalation schema. Drug was administered orally at escalating doses as a solution (25 to 850 mg bid) or as pellet capsules (500 to 1300 mg bid). Pharmacokinetics were assessed after the first dose and the last dose administered during cycle 1. RESULTS: Dose-limiting toxicity of grade 3 neuropathy was observed in one patient and grade 2 fatigue (decrease in two performance status levels) was seen in four of six patients treated with 1,300 mg bid. The most frequent clinical grade 2 or 3 adverse events in any cycle included nausea, vomiting, headache, fatigue, anemia, and hypotension. Myelosuppression was mild and infrequent. Peak plasma concentrations of R115777 were achieved within 0.5 to 4 hours after oral drug administration. The elimination of R115777 from plasma was biphasic, with sequential half-lives of about 5 hours and 16 hours. There was little drug accumulation after bid dosing, and steady-state concentrations were achieved within 2 to 3 days. The pharmacokinetics were dose proportional in the 25 to 325 mg/dose range for the oral solution. Urinary excretion of unchanged R115777 was less than 0.1% of the oral dose. One patient with metastatic colon cancer treated at the 500-mg bid dose had a 46% decrease in carcinoembryonic antigen levels, improvement in cough, and radiographically stable disease for 5 months. CONCLUSION: R115777 is bioavailable after oral administration and has an acceptable toxicity profile. Based upon pharmacokinetic data, the recommended dose for phase II trials is 500 mg orally bid (total daily dose, 1, 000 mg) for 5 consecutive days followed by 9 days of rest. Studies of continuous dosing and studies of R115777 in combination with chemotherapy are ongoing. PMID- 10673538 TI - The logic of evidence. PMID- 10673539 TI - Radiation for early-stage Hodgkin's disease? PMID- 10673540 TI - Swelling-activated, cystic fibrosis transmembrane conductance regulator-augmented ATP release and Cl- conductances in murine C127 cells. AB - 1. A hypotonic challenge, but not cAMP stimulation, was found to induce release of ATP measured by the luciferin-luciferase assay from both the murine mammary carcinoma cell line C127i and C127 cells stably transfected with the cDNA for human cystic fibrosis transmembrane conductance regulator (CFTR) protein (C127/CFTR). CFTR expression augmented swelling-induced ATP release by 10-20 times under hypotonic conditions (< or = 80 % osmolality). 2. Glibenclamide failed to suppress swelling-induced ATP release from C127/CFTR cells. In contrast, whole-cell patch-clamp recordings showed that both the cAMP-activated ohmic Cl- currents and volume-sensitive outwardly rectifying (VSOR) Cl- currents were prominently suppressed by glibenclamide. 3. Gd3+ markedly blocked swelling induced ATP release but failed to suppress both cAMP- and swelling-activated Cl- currents in the CFTR-expressing cells. Even after pretreatment and during treatment with Gd3+, VSOR Cl- currents were activated normally. 4. The continuous presence of an ATP-hydrolysing enzyme, apyrase, in the bathing solution did not prevent activation of VSOR Cl- currents in C127/CFTR cells. 5. The rate of regulatory volume decrease (RVD) in C127/CFTR cells was much faster than that in C127i cells. When apyrase was added to the bathing solution, the RVD rate was retarded in C127/CFTR cells. 6. On balance, the following conclusions can be deduced. First, swelling-induced ATP release is augmented by expression of CFTR but is not mediated by the CFTR Cl- channel. Second, swelling-induced ATP release is not mediated by the VSOR Cl- channel. Third, the released ATP facilitated the RVD process but is not involved in the activation of VSOR Cl- channels in C127/CFTR cells. PMID- 10673541 TI - Heterodimeric amino acid transporters: expression of heavy but not light chains of CD98 correlates with induction of amino acid transport systems in human placental trophoblast. AB - 1. Activity of amino acid transport and relative abundance of mRNAs encoding related transporters have been studied in parallel either before or following in vitro culture of explants of human placental chorionic villi. 2. Amino acid transport activities through systems L (1.9-fold), y+L (2.6-fold) and y+ (3.2 fold) were markedly enhanced following culture for 48 h. 3. Relative mRNA abundance (determined by reverse transcription-polymerase chain reaction) for the heavy chain of CD98 surface antigen and for the cationic amino acid transporter-1 were similarly stimulated (2.8-fold and 2.6-fold, respectively). In contrast, none of the mRNA levels for light chains of CD98 (system L-amino acid transporter 1, system L-amino acid transporter-2, system y+L-amino acid transporter-1 and system y+L-amino acid transporter-2) studied nor for the cationic amino acid transporter-2B were altered. PMID- 10673543 TI - Effects of cytosolic ATP on spontaneous and triggered Ca2+-induced Ca2+ release in permeabilised rat ventricular myocytes. AB - 1. The effects of cytosolic ATP on sarcoplasmic reticulum (SR) Ca2+ regulation were investigated in saponin-permeabilised rat ventricular myocytes. [Ca2+] within the cells was monitored using Fura-2 or Fluo-3 fluorescence. Spontaneous cyclic Ca2+ release from the SR was induced by increasing the bathing [Ca2+] to 200-300 nM, in solutions weakly Ca2+ buffered with 0.05 mM EGTA. Alternatively, Ca2+-induced Ca2+ release (CICR) was triggered by a rapid increase in [Ca2+] induced by flash photolysis of Nitr-5 (0.08 mM), replacing EGTA in the solution. 2. Stepwise reductions in [ATP] were associated with corresponding decreases in the frequency and increases in the amplitude of spontaneous Ca2+ transients. A decrease from 5 mM to 0. 1 mM ATP, reduced the release frequency by 48.6 +/- 7 % (n = 7) and almost doubled the amplitude of the Ca2+ transient. Marked prolongation of the spontaneous Ca2+ transient occurred when [ATP] was further reduced to 10 microM, consistent with inhibition of the SR Ca2+ pump. 3. These effects of ATP were compared with other interventions that inhibit Ca2+ uptake or reduce the sensitivity of the SR Ca2+ release mechanism. Inhibition of the SR Ca2+ pump with cyclopiazonic acid (CPA) markedly reduced the spontaneous Ca2+ release frequency, without changing the amplitude. The descending phase of the Ca2+ transient was prolonged in the presence of CPA, while the rising phase was unaffected. In contrast, desensitisation of the SR Ca2+ release mechanism with tetracaine decreased the frequency of spontaneous release, but markedly increased the amplitude. 4. CICR triggered by flash photolysis of Nitr-5 appeared to be more sensitive to cytosolic [ATP] than spontaneous release and was generally delayed by a decrease to 2.5 mM ATP. In the presence of 0.1-0.2 mM ATP, release often failed completely or was not consistently triggered. Some preparations exhibited Ca2+ release 'alternans', whereby every alternate trigger induced a response. 5. These results suggest that the increase in spontaneous Ca2+ release amplitude and the decrease in frequency that occurs as [ATP] is reduced from 1 mM to 100 microM, is mainly due to desensitisation of the SR Ca2+ release mechanism, which allows the SR Ca2+ content to reach a higher level before release occurs. At very low [ATP], a reduction in the SR Ca2+ uptake rate may also contribute to the decrease in release frequency. CICR triggered by photolysis of Nitr-5 appeared to be more sensitive to cytosolic [ATP]. The possible underlying mechanisms and the relevance of these results to myocardial ischaemia or hypoxia is considered. PMID- 10673542 TI - Reversible Ca2+-induced fast-to-slow transition in primary skeletal muscle culture cells at the mRNA level. AB - 1. The adult fast character and a Ca2+-inducible reversible transition from a fast to a slow type of rabbit myotube in a primary culture were demonstrated at the mRNA level by Northern blot analysis with probes specific for different myosin heavy chain (MyHC) isoforms and enzymes of energy metabolism. 2. No non adult MyHC isoform mRNA was detected after 22 days of culture. After 4 weeks of culture the fast MyHCIId mRNA was strongly expressed while MyHCI mRNA was virtually absent, indicating the fast adult character of the myotubes in the primary skeletal muscle culture. 3. The data show that a fast-to-slow transition occurred in the myotubes at the level of MyHC isoform gene expression after treatment with the Ca2+ ionophore A23187. The effects of ionophore treatment were decreased levels of fast MyHCII mRNA and an augmented expression of the slow MyHCI gene. Changes in gene expression started very rapidly 1 day after the onset of ionophore treatment. 4. Levels of citrate synthase mRNA increased and levels of glyceraldehyde 3-phosphate dehydrogenase mRNA decreased during ionophore treatment. This points to a shift from anaerobic to oxidative energy metabolism in the primary skeletal muscle culture cells at the level of gene expression. 5. Withdrawal of the Ca2+ ionophore led to a return to increased levels of MyHCII mRNA and decreased levels of MyHCI mRNA, indicating a slow-to-fast transition in the myotubes and the reversibility of the effect of ionophore on MyHC isoform gene expression. PMID- 10673544 TI - L-type calcium channel activity regulates sodium channel levels in rat pituitary GH3 cells. AB - 1. The effects of chronic pharmacological modulation of L-type Ca2+ channel activity on the cell surface expression of Na+ channels were examined in GH3 cells. 2. Prolonged inhibition (4-5 days) of L-channels with nimodipine caused a 50-60 % decrease in the peak amplitude of whole-cell Na+ currents recorded with the patch-clamp technique. On the contrary, prolonged exposure to the L-channel agonist Bay K 8644 induced an approximately 2.5-fold increase in peak Na+ current. In both cases, there were only minor changes in cell capacitance and no significant changes in Na+ channel gating properties. 3. Measurements of the specific binding of radiolabelled saxitoxin to intact cells showed that nimodipine treatment reduced the number of cell surface Na+ channels, whereas treatment with Bay K 8664 produced the opposite effect. The dual regulation of Na+ channel abundance explained the mentioned changes in Na+ current amplitude. 4. Plasma membrane Na+ channels had a half-life of approximately 17 h both in control cells and in cells treated with Bay K 8644, as estimated from the rate of decay of peak Na+ current after inhibition of protein synthesis with cycloheximide. Actinomycin D, an inhibitor of gene transcription, and also cycloheximide, occluded the stimulatory effect of Bay K 8644 on Na+ current density when measured over a 24 h period. 5. These findings indicate that the entry of Ca2+ through L-type channels influences in a positive way the number of functional Na+ channels in GH3 cells, and suggest that Ca2+ influx stimulates either Na+ channel gene expression or the expression of a regulatory protein that promotes translocation of pre-assembled Na+ channels into the plasma membrane. PMID- 10673545 TI - Ca2+ influx via the L-type Ca2+ channel during tail current and above current reversal potential in ferret ventricular myocytes. AB - 1. Current through L-type Ca2+ channels (ICa) was measured electrophysiologically at the same time as Ca2+ influx was measured by trapping entering Ca2+ with a high concentration of indo-1 (> 1 mM) in ferret ventricular myocytes. 2. Na+-free conditions prevented Na+-Ca2+ exchange and K+ currents were blocked by Cs+ and TEA. Thapsigargin (5 microM) prevented Ca2+ uptake and release by the sarcoplasmic reticulum. ICa was pre-activated by brief pulses to +120 mV (the equilibrium potential for Ca2+, ECa), followed by steps to different membrane potentials (Em, -80 to +100 mV), in some cases in the presence of the Ca2+ channel agonist FPL-64176. 3. Integrated ICa ( 82 ICa) was linearly related to the change in the concentration of Ca2+ bound to indo-1, which was assessed by the fluorescence difference signal DeltaFd (Fd = F500 - F400). This created an internal calibration of DeltaFd as a measure of Ca2+ influx. 4. The DeltaFd/ 82 ICadt relationship was virtually unchanged at all measurable inward ICa (at Em from -80 to +50 mV). This indicates that the fractional current carried by Ca2+ and channel selectivity are unchanged over this Em range, and also that the selectivity for Ca2+ is very high. 5. Ca2+ influx was readily detected by DeltaFd beyond the ICa reversal potential (+65 to +100 mV) and was not abolished until Em was +120 mV (i.e. ECa). This is explained by the fact that inward Ca2+ flux at the ICa reversal potential is exactly balanced by outward Cs+ current through the Ca2+ channels and can be described by classic Goldman flux analysis with a Ca2+/Cs+ selectivity of the order of 5000. 6. This result also emphasizes that net Ca2+ influx via Ca2+ channels occurs over a voltage range where the net channel current is outward. PMID- 10673546 TI - Calcium-, voltage- and osmotic stress-sensitive currents in Xenopus oocytes and their relationship to single mechanically gated channels. AB - 1. Patch recordings from Xenopus oocytes indicated that mechanically gated (MG) channels are expressed at a uniform surface density ( approximately 1 channel microm-2) with an estimated > 3 x 106 MG channels per oocyte that could generate microamps of current at +/-50 mV. 2. Removal of external Ca2+ induced a membrane conductance that differed from MG channels in ion selectivity, pharmacology and sensitivity to connexion-38. 3. Depolarization to +50 mV activated a Na+ selective, a Cl--selective and a non-selective conductance. Hyperpolarization to 150 mV activated a non-selective conductance. None of these conductances appeared to be mediated by MG channels. 4. Hypotonicity (25 %) failed to evoke any change in membrane conductance in the majority of defolliculated oocytes. Hypertonicity (200 %) evoked a large non-selective (PK /PCl approximately 1) membrane conductance that was not blocked by 100 microM Gd3+. 5. Although the above stimuli could activate a variety of whole-oocyte conductances, including three novel conductances, they did not involve MG channel activation. Possible mechanisms underlying the discrepancy between observed conductances and those anticipated from patch-clamp studies are discussed. PMID- 10673547 TI - On the discrepancy between whole-cell and membrane patch mechanosensitivity in Xenopus oocytes. AB - 1. Mechanical stimulation of voltage-clamped Xenopus oocytes by inflation, aspiration, or local indentation failed to activate an increase in membrane conductance up to the point of causing visible oocyte damage. 2. The absence of mechanosensitivity is not due to the vitelline membrane, rapid MG channel adaptation or tension-sensitive recruitment of new membrane. 3. Membrane capacitance measurements indicate that the oocyte surface area is at least 5 times larger than that predicted assuming a smooth sphere. We propose that this excess membrane area provides an immediate reserve that can 'buffer' membrane tension changes and thus prevent MG channel activation. 4. High-resolution images of tightly sealed patches and patch capacitance measurements indicate a smooth membrane that is pulled flat and perpendicular across the inside of the pipette. Brief steps of pressure or suction cause rapid and reversible membrane flexing and MG channel activation. 5. We propose that changes in membrane geometry induced during cell growth and differentiation or as a consequence of specific physiological and pathological conditions may alter mechanosensitivity of a cell independently of the intrinsic properties of channel proteins. PMID- 10673548 TI - Mechanically gated channel activity in cytoskeleton-deficient plasma membrane blebs and vesicles from Xenopus oocytes. AB - 1. A novel technique involving hypertonic stress causes membrane 'blebbing' of the Xenopus oocyte and the shedding of plasma membrane vesicles (PMVs). 2. Confocal fluorescence microscopy, immunocytochemistry and electron microscopy indicate that blebs and PMVs lack cortical cytoskeleton and are deficient in cytoskeleton proteins and devoid of microvilli. 3. Patch recordings from PMVs consistently reveal mechanically gated (MG) channel activity. The MG channels display the same single-channel conductance as control recordings but differ in terms of reduced mechanosensitivity and adaptation to sustained stimulation. 4. Whole PMV recordings show rapid and reversible activation of mechanosensitive currents in response to pressure pulses. The maximal currents activated in PMVs are consistent with MG channel activity recorded in patches. 5. The discrepancy between MG channel activity recorded in whole PMVs and oocytes most probably reflects their different membrane geometry and ability to develop activating bilayer tensions. 6. We propose that membrane blebbing, which is known to occur under specific physiological and pathological conditions (e.g. mitosis and apoptosis), may increase mechanosensitivity independently of the intrinsic properties of membrane proteins. PMID- 10673549 TI - Signalling pathway for histamine activation of non-selective cation channels in equine tracheal myocytes. AB - 1. The signalling pathway underlying histamine activation of non-selective cation channels was investigated in single equine tracheal myocytes. Application of histamine (100 microM) activated the transient calcium-activated chloride current (ICl(Ca)) and sustained, low amplitude non-selective cation current (ICat). The H1 receptor antagonist pyrilamine (10 microM) blocked activation of ICl(Ca) and ICat. Simultaneous application of histamine (100 microM) and caffeine (8 mM) during H1 receptor blockade activated ICl(Ca), but not ICat. Neither the H2 receptor antagonist cimetidine (20 microM) nor the H3 receptor antagonist thioperamide (20 microM) prevented activation of ICl(Ca) and ICat. 2. Intracellular dialysis of anti-Galphai/Galphao antibodies completely blocked activation of ICat by histamine, whereas ICl(Ca) was not affected. By contrast, anti-Galphaq/Galpha11 antibodies greatly inhibited ICl(Ca), but did not alter activation of ICat. 3. 1-Oleoyl-2-acetyl-sn-glycerol (OAG, 20-100 microM) did not induce any current or affect currents activated by histamine or methacholine (mACH). Simultaneous application of OAG and caffeine activated ICl(Ca), but not ICat, indicating that a rise in [Ca2+]i and stimulation of diacylglycerol sensitive protein kinase C (PKC) is not sufficient to activate ICat. The phospholipase C inhibitor U73122 (2 microM) blocked histamine activation of ICl(Ca) and ICat, but simultaneous exposure of myocytes to histamine and caffeine restored both ICl(Ca) and ICat in the presence of U73122. 4. Histamine and mACH activated currents with equivalent I-V relationships. The currents activated by these agonists were not additive; following activation of ICat by mACH, histamine failed to induce an additional membrane current. Similarly, mACH did not induce an additional current after full activation of ICat by histamine. 5. We conclude that H1 histamine receptors activate ICat through coupling to Gi/Go proteins. Activation of ICat also requires intracellular calcium release, mediated by H1 receptors coupling to Gq/G11 proteins. This coupling is analogous to the activation of ICat by co-stimulation of M2 and M3 receptors. PMID- 10673550 TI - Muscarinic receptor-induced acidification in sublingual mucous acinar cells: loss of pH recovery in Na+-H+ exchanger-1 deficient mice. AB - 1. Intracellular pH (pHi) plays an important role in regulating fluid and electrolyte secretion by salivary gland acinar cells. The pH-sensitive, fluorescent dye 2', 7'-bis(carboxyethyl)-5(6)-carboxylfluorescein (BCECF) was used to characterize the mechanisms involved in regulating pHi during muscarinic stimulation in mouse sublingual mucous acinar cells. 2. In the presence of HCO3-, muscarinic stimulation caused a rapid decrease in pHi (0.24 +/- 0.02 pH units) followed by a slow recovery rate (0.042 +/- 0.002 pH units min-1) to the initial resting pHi in sublingual acinar cells. The muscarinic receptor-induced acidification in parotid acinar cells was of a similar magnitude (0. 25 +/- 0.02 pH units), but in contrast, the recovery rate was approximately 4-fold faster (0.181 +/- 0.005 pH units min-1). 3. The agonist-induced intracellular acidification was inhibited by the anion channel blocker niflumate, and was prevented in the absence of HCO3- by treatment with the carbonic anhydrase inhibitor methazolamide. These results indicate that the muscarinic-induced acidification is due to HCO3- loss, probably mediated by an anion conductive pathway. 4. The Na+-H+ exchange inhibitor 5-(N-ethyl-N-isopropyl)amiloride (EIPA) amplified the magnitude of the agonist-induced acidification and completely blocked the Na+-dependent pHi recovery. 5. To examine the molecular nature of the Na+-H+ exchange mechanism in sublingual acinar cells, pH regulation was investigated in mice lacking Na+-H+ exchanger isoforms 1 and 2 (NHE1 and NHE2, respectively). The magnitude and the rate of pHi recovery in response to an acid load in acinar cells isolated from mice lacking NHE2 were comparable to that observed in cells from wild-type animals. In contrast, targeted disruption of the Nhe1 gene completely abolished pHi recovery from an acid load. These results demonstrate that NHE1 is critical for regulating pHi during a muscarinic agonist stimulated acid challenge and probably plays an important role in regulating fluid secretion in the sublingual exocrine gland. 6. In NHE1-deficient mice, sublingual acinar cells failed to recover from an acid load in the presence of bicarbonate. These results confirm that the major regulatory mechanism involved in pHi recovery from an acid load is not Na+-HCO3- cotransport, but amiloride sensitive Na+-H+ exchange via isoform 1. PMID- 10673551 TI - Cell volume regulation: the role of taurine loss in maintaining membrane potential and cell pH. AB - 1. In response to a hypo-osmotic stress cells undergo a regulatory volume decrease (RVD) by losing osmotically active solutes and obliged water. During RVD, trout red cells lost taurine, K+ and Cl- but gained Na+ and Cl-. Over the full time course of RVD the chloride concentration in the cell water remained remarkably constant. Thus membrane potential and cell pH, which depends on the ratio of internal to external chloride concentration ([Cl-]i:[Cl-]o), remained fixed. 2. When cell volume decreases it is only possible to keep the chloride concentration in the cell water constant if an equal percentage of the cell chloride pool and of the cell water pool are lost simultaneously. Quantitative analysis of our data showed that this requirement was fulfilled because, over the full time course of RVD, cells lost osmotically active solutes with a constant stoichiometry: 1 Cl-:1 positive charge:2.35 taurine. Any change in taurine permeability, by modifying the stoichiometric relationship, would affect the amount of water lost and consequently cell chloride concentration. 3. Experiments carried out with different cations as substitutes for external Na+ suggest that the constancy of the chloride concentration is not finely tuned by some mechanism able to modulate the channel transport capacity, but results in part from the fact that the swelling-dependent channel constitutively possesses an adequately fixed relative permeability for cations and taurine. However, as a significant fraction of K+ and Cl- loss occurs via a KCl cotransporter, the contribution of the cotransport to the stoichiometric relationship remains to be defined. 4. The large amount of taurine released during RVD (50 % of all solutes) was shown to be transported as an electroneutral zwitterion and not as an anion. How the channel can accommodate the zwitterionic form of taurine, which possesses a high electrical dipole, is considered. PMID- 10673552 TI - Effect of hypothermia on the volume of rat glial cells. AB - 1. The cell volume of suspended C6 glioma cells and primary cultured rat astrocytes was measured at normothermia (37 degrees C), and at mild (32 degrees C) and moderate (27 degrees C) hypothermia by flow cytometry with electrical cell sizing. 2. Under control conditions (37 degrees C), C6 glioma cells had a volume of 809 +/- 29 microm3. Moderate hypothermia (27 degrees C) led to rapid cell swelling, with a maximum volume of 113.1 +/- 1.3 % of control being achieved after 50 min. After rewarming to 37 degrees C, cell volume recovered very slowly and incompletely (to 107.2 +/- 0.4 % of control). Less severe hypothermia (32 degrees C) led to a smaller increase in cell volume (108.7 +/- 0.5 % of control). 3. The maximal cell swelling response and the kinetics of swelling were similar in C6 glioma cells and primary cultured astrocytes. 4. Hypothermia-induced cell swelling was dependent on the presence of extracellular Na+ and was reduced by the Na+-H+ antiporter inhibitor EIPA. 5. The underlying mechanisms of hypothermia induced cell swelling are an intracellular accumulation of Na+ by (1) differential effects of hypothermia on the membrane permeabilities of Na+ and K+ and (2) activation of the Na+-H+ antiporter by a shift of its activation curve to a more alkaline value. PMID- 10673554 TI - Weak common parallel fibre synapses explain the loose synchrony observed between rat cerebellar golgi cells. AB - 1. In anaesthetized rats, pairs of cerebellar Golgi cells fired synchronously at rest, provided they were aligned along the parallel fibre axis. The observed synchrony was much less precise, however, than that which would be expected to result from common, monosynaptic parallel fibre excitation. 2. To explain this discrepancy, the precision and frequency of spike synchronization (i.e. the width and area of the central peak on the spike train cross-correlogram) were computed in a generic model for varying input, synaptic and neuronal parameters. 3. Correlation peaks between model neurons became broader, and peak area smaller, when the number of afferents increased and each single synapse decreased proportionally in strength. Peak width was inversely proportional to firing rate, but independent of the percentage of shared afferents. Peak area, in contrast, scaled with the percentage of shared afferents but was almost firing rate independent. 4. Broad correlation peaks between pairs of model neurons resulted from the loose spike timing between single model neurons and their afferents. This loose timing reflected a need for long-term synaptic integration to fire the neurons. Model neurons could accomplish this through firing rate adaptation mediated by a Ca2+-activated K+ channel. 5. We conclude that loose synchrony may be entirely explained by shared input from monosynaptic, non-synchronized afferents. The inverse relationship between peak width and firing rate allowed us to distinguish common parallel fibre input from firing rate covariance as a primary cause of loose synchrony between cerebellar Golgi cells in anaesthetized rats. PMID- 10673553 TI - Presynaptic dopamine D2-like receptors inhibit excitatory transmission onto rat ventral tegmental dopaminergic neurones. AB - 1. The effects of dopamine (DA) on non-NMDA glutamatergic transmission onto dopaminergic neurones in the ventral tegmental area (VTA) were examined in rat midbrain slices using the whole-cell patch-clamp technique. EPSCs in dopaminergic neurones evoked by focal stimulation within the VTA were reversibly blocked by 5 microM CNQX in the presence of bicuculline (20 microM), strychnine (0.5 microM) and D-amino-5-phosphonopentanoic acid (D-AP5, 25 microM). 2. Bath application of DA reduced the amplitude of EPSCs up to 65.1 +/- 9. 52% in a concentration dependent manner between 0.3-1000 microM (IC50, 16.0 microM) without affecting the holding current at -60 mV measured using a Cs+-filled electrode. 3. The effect of DA on evoked EPSCs was mimicked by the D2-like receptor agonist quinpirole but not by the D1-like receptor agonist SKF 81297, and was antagonized by the D2-like receptor antagonist sulpiride (KB, 0.96 microM), but not by the D1 like receptor antagonist SCH 23390 (KB, 228.6 microM). 4. Dopamine (30 microM) reduced the mean frequency of spontaneous miniature EPSCs (mEPSCs) without affecting their mean amplitude, and the DA-induced effect on the mEPSCs was dependent on the external Ca2+ concentration. 5. These results suggest that afferent glutamatergic fibres which terminate on VTA dopaminergic neurones possess presynaptic D2-like receptors, activation of which inhibits glutamate release by reducing Ca2+ influx. PMID- 10673555 TI - Voltage-gated currents distinguish parvocellular from magnocellular neurones in the rat hypothalamic paraventricular nucleus. AB - 1. Magnocellular and parvocellular neurones of the hypothalamic paraventricular nucleus (PVN) differentially regulate pituitary hormone secretion and autonomic output. Previous experiments have suggested that magnocellular, or type I neurones, and parvocellular, or type II neurones, of the PVN express different electrophysiological properties. Whole-cell patch-clamp recordings were performed in hypothalamic slices to identify the voltage-gated currents responsible for the electrophysiological differences between type I and type II PVN neurones. 2. Type I neurones, which display transient outward rectification and lack a low threshold spike (LTS), generated a large A-type K+ current (IA) (mean +/- s.e. m.: 1127.5 +/- 126.4 pA; range: 250-3600 pA; voltage steps to -25 mV) but expressed little or no T-type Ca2+ current (IT). Type II neurones, which lack transient outward rectification but often display an LTS, expressed a smaller IA (360.1 +/- 56.3 pA; range: 40-1100 pA; voltage steps to -25 mV), and 75 % of the type II neurones generated an IT (-402.5 +/- 166.9 pA; range: -90 to -2200 pA; at peak). 3. The voltage dependence of IA was shifted to more negative values in type I neurones compared to type II neurones. Thus, the activation threshold ( 53.5 +/- 0.9 and -46.1 +/- 2.6 mV), the half-activation potential (-25 +/- 1.9 and -17.9 +/- 2.0 mV), the half-inactivation potential (-80.4 +/- 9.3 and -67.2 +/- 3.0 mV), and the potential at which the current became fully inactivated ( 57.4 +/- 2.1 and -49.8 +/- 1.5 mV) were more negative in type I neurones than in type II neurones, respectively. 4. IT in type II neurones activated at a threshold of -59.2 +/- 1.2 mV, peaked at -32. 6 +/- 1.7 mV, was half-inactivated at -66.9 +/- 2.2 mV, and was fully inactivated at -52.2 +/- 2.2 mV. 5. Both cell types expressed a delayed rectifier current with similar voltage dependence, although it was smaller in type I neurones (389.7 +/- 39.3 pA) than in type II neurones (586.4 +/- 76.0 pA). 6. In type I neurones IA was reduced by 41.1 +/- 7.0 % and the action potential delay caused by the transient outward rectification was reduced by 46.2 +/- 10.3 % in 5 mM 4-aminopyridine. In type II neurones IT was reduced by 66.8 +/- 10.9 % and the LTS was reduced by 76.7 +/- 7.8 % in 100 microM nickel chloride, but neither IT nor LTS was sensitive to 50 microM cadmium chloride. 7. Thus, differences in the electrophysiological properties between type I, putative magnocellular neurones and type II, putative parvocellular neurones of the PVN can be attributed to the differential expression of voltage-gated K+ and Ca2+ currents. This diversity of ion channel expression is likely to have profound effects on the response properties of these neurosecretory and non-neurosecretory neurones. PMID- 10673556 TI - Responses of neurones of the rat suprachiasmatic nucleus to retinal illumination under photopic and scotopic conditions. AB - 1. We have examined the responses of neurones in the suprachiasmatic nuclei (SCN) of the rat to retinal illumination under photopic and scotopic conditions to identify the types of photoreceptor input to these nuclei. 2. The majority of visually responsive SCN neurones studied under dark adaptation received rod input (48 of 52, 92 %). The action spectrum conformed to the sensitivity of rhodopsin, with maximal sensitivity at around 505 nm. 3. When also studied under light adaptation, most visually responsive SCN neurones (20 out of 26, 77 %) responded to input from cones. The action spectra conformed to the spectrum of green cone opsin, with a main sensitivity peak at 510 nm and a significant secondary peak in the near-ultraviolet region of the spectrum. 4. The frequency of spontaneous activity was typically low under scotopic conditions (range 0.2-17.2 Hz) and higher under photopic conditions (range 0.6-40 Hz) for any given neurone. The most common response under scotopic conditions was an 'on-excitation' (32 of 48, 62.5 %), which changed under photopic conditions to an on-excitation followed by a more prominent off-inhibition. 5. Responses also changed due to endogenous ultradian cycles. Depending on the phase, responses could be altogether absent and even reverted from excitation to inhibition on opposite phases of a cycle. Ultradian cycles had a circadian dependence and were most common at around the light phase:dark phase (L:D) and D:L transition points of the circadian cycle. 6. Under photopic conditions, SCN neurones showed rhythmic electrical activity, with a preferred firing interval that had a value between 18 and 39 ms. This rhythmic activity was probably the result of endogenous subthreshold membrane potential oscillations. 7. In conclusion, light acting either via rod or cone pathways could have powerful, opposing actions on SCN neurones. These actions were state dependent. The presence of these neuronal responses suggests a role for rod and cone photoreceptors in SCN function. PMID- 10673558 TI - Temporal coupling between neuronal activity and blood flow in rat cerebellar cortex as indicated by field potential analysis. AB - 1. Laser-Doppler flowmetry and extracellular recordings of field potentials were used to examine the temporal coupling between neuronal activity and increases in cerebellar blood flow (CeBF). 2. Climbing fibre-evoked increases in CeBF were dependent on stimulus duration, indicating that increases in CeBF reflected a time integral in neuronal activity. The simplest way to represent neuronal activity over time was to obtain a running summation of evoked field potential amplitudes (runSigmaFP). RunSigmaFP was calculated for each stimulus protocol and compared with the time course of the CeBF responses to demonstrate coupling between nerve cell activity and CeBF. 3. In the climbing fibre system, the amplitude and time course of CeBF were in agreement with the calculated postsynaptic runSigmaFP (2-20 Hz for 60 s). This suggested coupling between CeBF and neuronal activity in this excitatory, monosynaptic, afferent-input system under these conditions. There was no correlation between runSigmaFP and CeBF during prolonged stimulation. 4. Parallel fibre-evoked increases in CeBF correlated with runSigmaFP of pre- and postsynaptic potentials (2-15 Hz for 60 s). At higher stimulation frequencies and during longer-lasting stimulation the time course and amplitudes of CeBF responses correlated with runSigmaFP of presynaptic, but not postsynaptic potentials. This suggested a more complex relationship in this mixed inhibitory-excitatory, disynaptic, afferent-input system. 5. This study has demonstrated temporal coupling between neuronal activity and CeBF in the monosynaptic, excitatory climbing-fibre system. In the mixed mono- and disynaptic parallel fibre system, temporal coupling was most clearly observed at low stimulation frequencies. We propose that appropriate modelling of electrophysiological data is needed to document functional coupling of neuronal activity and blood flow. PMID- 10673557 TI - Chronic passive cigarette smoke exposure augments bronchopulmonary C-fibre inputs to nucleus tractus solitarii neurones and reflex output in young guinea-pigs. AB - 1. Children chronically exposed to environmental tobacco smoke (passive cigarette smoke) have more wheeze, cough, bronchoconstriction, airway hyper-reactivity and mucous secretion, which may result, in part, from stimulation of the vagal bronchopulmonary C-fibre reflex. 2. Environmental tobacco smoke increases the sensitivity of bronchopulmonary C-fibre endings, but the physiological relevance of this sensitization is unknown. If this exposure augments the reflex responses via a central mechanism, then the responses of higher-order neurones in the reflex pathway and some components of the reflex output should also be augmented. 3. Guinea-pigs were chronically exposed to sidestream tobacco smoke (surrogate for environmental tobacco smoke) or filtered air for 5 days week-1 from age 1 to 6 weeks (age equivalent of human childhood) and were then anaesthetized, paralysed, ventilated and prepared with pneumothoraces. Baseline and left atrial capsaicin (0.5 and 2.0 microg kg-1)- evoked changes in the impulse activity of vagal C-fibre-activated neurones in nucleus tractus solitarii (NTS), phrenic nerve activity, tracheal pressure, arterial blood pressure and heart rate were compared in the two groups. 4. Sidestream smoke exposure significantly augmented the peak (P = 0.02) and duration (P = 0.01) of the NTS neuronal responses and the prolongation of expiratory time (P = 0.003) at the higher capsaicin dose. 5. Thus, the sensitization of the bronchopulmonary C-fibre endings by chronic exposure to sidestream tobacco smoke is transmitted to the NTS and is associated with a prolonged reflexively evoked expiratory apnoea. The findings may help to explain some related respiratory symptoms in children and be a factor in sudden infant death syndrome. PMID- 10673559 TI - Recycling and refilling of transmitter quanta at the frog neuromuscular junction. AB - 1. Fluorescent dyes have been used at the frog neuromuscular junction to label synaptic vesicular membrane. Retrieved membrane is reformed into vesicles, which are released along with pre-existing vesicles. Consequently, if vesicular refilling with acetylcholine (ACh) is depressed by inhibitors, two sizes of quanta should be released: normal and smaller. As recycling continues the fraction of smaller size quanta should increase exponentially. 2. We enhanced the rate of quantal release by elevating the K+ concentration. The principal inhibitors were (-)-vesamicol (VES), hemicholinium-3 (HC3), and NH4+. Quantal size measurements were fitted to one and to two cumulative lognormal probability distribution functions. When two fitted better, the statistical significance assessment took into account the three additional parameters used in calculating the fit. 3. After recycling in the presence of inhibitor, many sets were fitted better by two lognormal functions. As recycling continued, the fraction of the miniature endplate potential voltage-time integrals ( MEPPs) in the larger sub population decreased exponentially. 4. The size of the releasable pool was estimated by counting the quanta released by carbonyl cyanide m chlorophenylhydrazone (CCCP). This was compared to pool sizes calculated from the inhibitor experiments. The two estimates of pool size were indistinguishable, with mean values ranging from about 170,000 to 270,000. 5. With all of the treatments tested, the means of the sizes in the smaller sub-population of MEPPs were about 1/3 those of the larger sub-populations. 6. Recycling synaptic vesicles appear to be incorporated into the releasable pool from which they have roughly the same probability of release as the pre-existing vesicles. PMID- 10673560 TI - Cerebral correlates of autonomic cardiovascular arousal: a functional neuroimaging investigation in humans. AB - 1. States of peripheral autonomic arousal accompany emotional behaviour, physical exercise and cognitive effort, and their central representation may influence decision making and the regulation of social and emotional behaviours. However, the cerebral functional neuroanatomy representing and mediating peripheral autonomic responses in humans is poorly understood. 2. Six healthy volunteer subjects underwent H215O positron emission tomography (PET) scanning while performing isometric exercise and mental arithmetic stressor tasks, and during corresponding control tasks. Mean arterial blood pressure (MAP) and heart rate (HR) were monitored during scanning. 3. Data were analysed using statistical parametric mapping (SPM99). Conjunction analyses were used to determine significant changes in regional cerebral blood flow (rCBF) during states of cardiovascular arousal common to both exercise and mental stressor tasks. 4. Exercise and mental stressor tasks, relative to their control tasks, were associated with significantly (P < 0.001) increased MAP and HR. Significant common activations (increased rCBF) were observed in cerebellar vermis, brainstem and right anterior cingulate. In both exercise and mental stress tasks, increased rCBF in cerebellar vermis, right anterior cingulate and right insula covaried with MAP; rCBF in pons, cerebellum and right insula covaried with HR. Cardiovascular arousal in both categorical and covariance analyses was associated with decreased rCBF in prefrontal and medial temporal regions. 5. Neural responses in discrete brain regions accompany peripheral cardiovascular arousal. We provide evidence for the involvement of areas previously implicated in cognitive and emotional behaviours in the representation of peripheral autonomic states, consistent with a functional organization that produces integrated cardiovascular response patterns in the service of volitional and emotional behaviours. PMID- 10673561 TI - Increased muscle spindle sensitivity to movement during reinforcement manoeuvres in relaxed human subjects. AB - 1. The effects of reinforcement manoeuvres, such as mental computation and the Jendrassik manoeuvre, on muscle spindle sensitivity to passively imposed sinusoidal stretching (1.5 deg, 2 Hz) in relaxed subjects were analysed. 2. The unitary activity of 26 muscle spindle afferents (23 Ia, 3 II) originating from ankle muscles was recorded using the microneurographic method. Particular care was paid to the subjects' state of physical and mental relaxation. 3. The results showed that the activity of 54 % of the Ia afferents was modified during mental computation. The modifications took the form of either an increase in the number of spikes (mean, 26 % among 11 Ia fibres) or a shortening in the latency of the response to sinusoidal stretching (mean, 13 ms among 3 Ia fibres), or both. They were sometimes accompanied by an enhanced variability in the instantaneous discharge frequency. The three secondary endings tested exhibited no change in their sensitivity to stretch during mental computation. 4. The increased sensitivity to passive movements sometimes began as soon as the instructions were given to the subjects and sometimes increased during mental computation. In addition, the increased sensitivity either stopped after the subjects gave the right answer or continued for several minutes. 5. During the performance of a Jendrassik manoeuvre, the Ia units underwent changes similar to those described above for mental computation. 6. It was concluded that muscle spindle sensitivity to movement can be modified in relaxed human subjects. The results reinforce the idea that the fusimotor system plays a role in arousal and expectancy, and contribute to narrowing the gap between human and behaving animal data. PMID- 10673562 TI - Care of the child or adolescent with type 1 diabetes. AB - Type 1 diabetes in children and adolescents is a complex disease requiring multifaceted care. Children are managed with a combination of insulin by injection or pump, meal planning, and blood glucose self-monitoring. Close follow up by a pediatric diabetes treatment team is recommended for most children. Concerns related to short- and long-term complications and the complexity of day to-day management comprise the difficulties reported by parents and their children with type 1 diabetes. PMID- 10673564 TI - Management of nocturnal enuresis. AB - Heath care providers can assist the children and families who struggle with the problem of nocturnal enuresis in various ways. The children and their families need to receive adequate and appropriate information to assist with the psychosocial, emotional, and developmental issues involved. They also need to receive an adequate physical evaluation to diagnose or rule out diseases or conditions that can cause bedwetting. By taking on these tasks, advanced practice nurses can thoroughly and safely manage the care of the large population of children with nocturnal enuresis. PMID- 10673563 TI - Gastroesophageal reflux in children: a guide for the advanced practice nurse. AB - Gastroesophageal reflux (GER) is one of the most common gastrointestinal problems in infants and children. Unfortunately, the diagnosis of GER is often made after development of serious complications, placing the child at medical and developmental risk. Early recognition by primary care providers is essential in preventing these serious, sometimes life-threatening complications of untreated GER. This article reviews the pathophysiology, clinical manifestations, diagnostics, and treatment modalities of GER along with developmental considerations. As a health care provider, the advanced practice nurse is in an ideal position to provide a complete assessment, develop a realistic plan of care, coordinate interventions, assess outcomes, and offer support to the family of the child with GER. PMID- 10673565 TI - Identifying common pediatric neurosurgical conditions in the primary care setting. AB - The primary care setting is crucial to the diagnosis and treatment of pediatric neurosurgical conditions. The scope of this article is to present common pediatric neurosurgical diagnoses and their typical presentation to assist the primary care advanced practice nurse in making timely and appropriate referrals. The complications from delayed treatment can be significant for the child's future development. Conditions that are addressed include cutaneous stigmata related to occult spinal cord defects, common birth-related trauma, craniosynostosis, positional molding, torticollis, hydrocephalus, mild head injury, and brain tumors. PMID- 10673566 TI - The judicious use of antibiotic agents in common childhood respiratory illness. AB - Increased bacterial resistance is caused most frequently by the widespread use of antimicrobial agents. Antimicrobial agents are often used inappropriately to treat common respiratory illnesses in children. This article discusses the judicious use of antimicrobials in the common cold, otitis media, acute sinusitis, pharyngitis, and bronchitis. PMID- 10673567 TI - Practical primary pediatric orthopedics. AB - Pediatric orthopedic problems often puzzle the primary health provider and worry families. A thorough orthopedic history and examination by the primary care provider is all that is necessary to determine whether a problem requires further evaluation and referral. This article addresses a practical approach to common orthopedic problems, assessment, and management strategies from the specialist perspective. A general description of developmental dysplasia of the hip, foot misalignments, tibial torsion, toe-walking, genu varum (bowlegs), growing pains, sprains and fractures, and the child with a limp is provided. Management strategies before and after orthopedic referral are presented. PMID- 10673568 TI - Advanced practice nursing for children with HIV infection. AB - Children from any ethnic background and in any geographical location can be affected by HIV disease. Once acquired, HIV disease holds a devastating future for the infected child, his or her parents, and the entire family. As a multisystem disease, HIV affects every aspect of the child's growth and development. Management and prevention presents many challenges to the pediatric advanced practice nurse in the ambulatory care setting. This article reviews the epidemiology and transmission of HIV, as well as the clinical presentation of the HIV-infected child in the clinic. PMID- 10673569 TI - Advanced practice nursing in pediatric nephrology. AB - This article provides the nurse practitioner with a concise and accurate reference for the evaluation of children with hematuria, proteinuria, and hypertension. Although the incidence of these disorders in children is lower than in adults, they are not uncommon. The nurse practitioner must be able to accurately assess the results of a urinalysis and measure and interpret blood pressure readings in children. A discussion of when to refer children with signs and symptoms of serious disease; and how to educate patients and families in order to promote children's health is also provided. PMID- 10673570 TI - Pediatric febrile seizures and childhood headaches in primary care. AB - Febrile seizures and migraine headaches in children are two of the most common neurological diagnoses seen by primary care practitioners. It is essential that a knowledge base be developed to better care for this population. This article reviews pediatric febrile seizures, including management and treatment recommendations and childhood headaches, with an emphasis on migraine headaches. Diagnosis, management, and referral criteria are also reviewed. PMID- 10673571 TI - Pediatric dermatology: that itchy scaly rash. AB - Primary care practitioners in the pediatric setting treat children for numerous skin complaints. The most frequently seen dermatologic conditions are those that are persistent and cause children discomfort, such as atopic, contact seborrheic dermatitis and tinea infections. Familiarity with the presentation, pathophysiology, and treatment of these common skin conditions enables the practitioner to successfully manage these rashes. PMID- 10673572 TI - The role of the advanced practice nurse in pediatric general surgery. AB - The advanced practice nurse (APN) functions in pediatric surgery in a variety of ways. In the inpatient setting, the APN can provide care to infants and children with many complex congenital and other surgical anomalies. In the outpatient office, the role includes diagnosis, management, education, and care coordination for children with both routine and complex diagnoses. This article discusses the differences in some of these roles and a few of the common diagnoses seen by the APN in a pediatric surgical practice on a routine day. PMID- 10673573 TI - Immune mechanisms in rheumatic disease. AB - The pathophysiology of rheumatic diseases is complex and lacks clear-cut origins. This article describes the current theories and supporting evidence for the pathogenesis, mechanisms, and progression of rheumatic diseases. The roles of genetic inheritance and inflammatory/immune mediators are emphasized. PMID- 10673574 TI - Issues in the nursing management of osteoporosis. AB - As new interventions are developed to improve the care of patients with osteoporosis, nurses will play an important role in improving patients' quality of life, reducing their fear, and assisting in maintaining their independence throughout the adult life span. This article examines key issues involved in the management of patients at risk for, or diagnosed with, osteoporosis. PMID- 10673575 TI - Osteoarthritis: manageable scourge of aging. AB - Osteoarthritis is one of the most prevalent diseases of aging. An overview of osteoarthritis including definition, joints affected, clinical course, and classification is provided. Contributing factors, such as age, sex, ethnicity, obesity, diet, estrogen, and activity are discussed. Nursing interventions encompass weight control, adequate nutrition, evaluation of hormone replacement therapy, exercise and muscle strengthening, the use of assistive devices, medication assessment, pain control, psychosocial support, and the use of alternative therapies. Osteoarthritis can be successfully managed with the use of multifocal modalities. PMID- 10673576 TI - The role of exercise in the prevention and treatment of osteoporosis and osteoarthritis. AB - Osteoporosis and osteoarthritis are two distinctly different rheumatic conditions that target elderly, primarily female, populations. This article examines the scientific evidence supporting the use of exercise as a specific therapeutic modality, the general physiologic and psychological benefits of exercise, and the exercise programs currently recommended to combat these prevalent musculoskeletal disorders. Exercise is a valuable adjunct to treatment programs aimed at alleviating the risks and symptoms of osteoporosis and osteoarthritis. In addition to its potential impact on the disease processes themselves, exercise improves general health and well being, enhances quality of life, and preserves physical independence. PMID- 10673577 TI - Nonsteroidal anti-inflammatory drugs: new perspectives on a familiar drug class. AB - Nonsteroidal anti-inflammatory drugs are among the most commonly used and lethal drug classes. The anti-inflammatory, analgesic, and antipyretic effects result from the inhibition of cyclooxygenase (COX), which is the critical enzyme in the synthesis of prostaglandins. The most common adverse effects are gastrointestinal, but renal and platelet effects are also important. Recent discovery that there are two forms of cyclooxygenase (COX-1 and COX-2) has led to hypothesis that the newly marketed COX-2 selective inhibitor can provide beneficial effects without these adverse effects. Patient monitoring and education are nursing functions essential to the safe use of these agents. PMID- 10673578 TI - Polymyalgia rheumatica and temporal arteritis: a case presentation. AB - Polymyalgia rheumatica (PMA) and temporal arteritis (TA) are common clinical syndromes that affect the elderly population. Both syndromes may include similar constitutional complaints, an increase in acute phase reactants, a rapid response to corticosteroids, and the presence of anteritis giant cells upon temporal artery biopsy. Differential diagnosis is one of exclusion. A case presentation is included to assist practitioners in recognizing presenting symptoms, identifying diagnostic testing, and proceeding with appropriate therapy and follow-up. PMID- 10673579 TI - Systemic lupus erythematosus: a multisystem autoimmune disorder. AB - Systemic lupus erythematosus (SLE) is a chronic illness that has no known cause. As an autoimmune, inflammatory disease, it can affect various organs in different ways. Because of its fluctuating, unpredictable qualities, it requires the implementation of physiologic and psychosocial interventions that may change daily. A plan of care unique to each individual with SLE is essential to its management. PMID- 10673580 TI - Recognition and management of Sjogren's syndrome: strategies for the advanced practice nurse. AB - Sjogren's syndrome is an autoimmune disorder characterized by lymphocytic infiltration of salivary and lacrimal glands. The systemic production of autoantibodies leads to dry eyes, dry mouth, and other symptoms related to decreased salivary and lacrimal gland function in patients with Sjogren's syndrome. This article discusses origins, epidemiology, contributing factors, symptoms, assessment, diagnostic studies, management, expected outcomes, and research concerning Sjogren's syndrome. PMID- 10673581 TI - The psychosocial aspects of osteoporosis in women. AB - Osteoporosis has been documented to be a physiologically and psychologically debilitating disease. Health perceptions can be improved through both psychosocial support and specific intervention programs. These programs can improve independence and the quality of life of many people afflicted with this disease. PMID- 10673582 TI - Laboratory aspects of rheumatologic disease. AB - Assessment of laboratory values is an important function of nursing practice. Rheumatologic laboratory assessment, in particular, can be complex because few findings are actually pathognomonic. This article provides a perspective on an interpretive approach to laboratory assessment of rheumatologic disease. In conjunction with the patient's clinical status, these values can provide helpful information for monitoring or predicting the course of disease. PMID- 10673583 TI - [Biometrics and epidemiology]. PMID- 10673584 TI - Analysis of incomplete public health data. AB - The problem of dealing with missing values is common throughout statistics and is very prominent with epidemiologic data in the broad sense. Not only do data collection procedures break down, but subjects may be lost to follow up, or simply withdraw their consent without further providing a reason for doing so. In this paper, we review a framework for handling incomplete studies, and then concentrate on a specific case. It comes from a complex health interview survey, conducted in Belgium in 1997, where different types of missingness arise at various levels of the hierarchical sampling procedure. PMID- 10673585 TI - [Dealing with missing, abnormal and incoherent data in E3N cohort study]. AB - BACKGROUND: The E3N Study, 'Etude Epidemiologique aupres de femmes de la Mutuelle Generale de l'Education Nationale', is a cohort study, aiming at studying cancer risk factors on 100,000 women. Even if the incidence of problematic (missing, incoherent, etc.) data is low, any multivariate analysis which would be based only on complete subjects would rely on a too small sample, which would not necessarily be representative of the studied population. Results could thus be biased. METHODS: Our dealing with problematic data includes RESULTS: We looked at the number of individuals on which an analysis on 19 variables could be undertaken. The management of missing data made exploitable one fourth of the cohort, i.e.74.6% of individuals instead of 50.5%. Moreover, for 89.0% of subjects, one variable at most (out of the 19 studied) has missing datum. CONCLUSIONS: The main difficulty does not stand so much in the choice and implementation of methods to deal with problematic data than in the identification of their process of existence. Most of what was gained was due to the simplest methods: cold-deck and deductive method. PMID- 10673586 TI - [Analysis of longitudinal Gaussian data with missing data on the response variable]. AB - BACKGROUND: Using an application and a simulation study we show the bias induced by missing data in the outcome in longitudinal studies and discuss suitable statistical methods according to the type of missing responses when the variable under study is gaussian. METHOD: The model used for the analysis of gaussian longitudinal data is the mixed effects linear model. When the probability of response does not depend on the missing values of the outcome and on the parameters of the linear model, missing data are ignorable, and parameters of the mixed effects linear model may be estimated by the maximum likelihood method with classical softwares. When the missing data are non ignorable, several methods have been proposed. We describe the method proposed by Diggle and Kenward (1994) (DK method) for which a software is available. This model consists in the combination of a linear mixed effects model for the outcome variable and a logistic model for the probability of response which depends on the outcome variable. RESULTS: A simulation study shows the efficacy of this method and its limits when the data are not normal. In this case, estimators obtained by the DK approach may be more biased than estimators obtained under the hypothesis of ignorable missing data even if the data are non ignorable. Data of the Paquid cohort about the evolution of the scores to a neuropsychological test among elderly subjects show the bias of a naive analysis using all available data. Although missing responses are not ignorable in this study, estimates of the linear mixed effects model are not very different using the DK approach and the hypothesis of ignorable missing data. CONCLUSION: Statistical methods for longitudinal data including non ignorable missing responses are sensitive to hypotheses difficult to verify. Thus, it will be better in practical applications to perform an analysis under the hypothesis of ignorable missing responses and compare the results obtained with several approaches for non ignorable missing data. However, such a strategy requires development of new softwares. PMID- 10673587 TI - [Modeling correlated data in epidemiology: mixed or marginal model?]. AB - Correlated observations (within centers, families, subjects,.) are common in epidemiology. Even when one is only interested in the modeling of means according to risk factors, it is also necessary to model the variance-covariance matrix of the observations in order to make correct inferences on the parameters of interest. All the more so when the aim of the survey is the measurement of these correlations or of the variance of the random effects from which they are assumed to originate. We discuss, within the framework of the linear and of the logistic models, the implications of two choices for the modeling of covariances. The mixed model shows the unobserved elements responsible for the similarity between certain observations. In a longitudinal survey, for instance, one can use a random effect, specific to each subject, expressing how much a subject's trajectory is translated as compared to what is expected according to its characteristics (age, sex,.). The marginal approach leads to modeling separately the means and the covariance matrix of the observations. The distinction between these two approaches is important for non linear models, in particular the logistic one. We insist on the interconnection between a mixed model formulation and a marginal one, as well as on the implication of the choice in terms of the parameters' interpretation. PMID- 10673588 TI - [Methodology for analyzing censored correlated data: application of marginal and frailty approaches in human genetics. The European Community Alport Syndrome Concerted Action Group (ECASCA)]. AB - BACKGROUND: Statistical analysis for correlated censored data allows to study censored events in clustered structure designs. Considering a possible correlation among failure times of the same group, standard methodology is no longer applicable. We investigated proposed models in this context to study familial data about a genetic disease, Alport syndrome. Alport syndrome is a severe hereditary disease due to abnormal collagenous chains. Renal failure is the main symptom of the disease. It progresses toward end-stage renal failure (IRT) according to a high time variability. As shown by genetic studies, mutations of COL4A5 gene are involved in the X-linked Alport Syndrome. Due to the large range of the mutation types, the aim of this study was to search for a possible genetic origin of the heterogeneity of the disease severity. METHODS: Marginal survival models and mixed effects survival models (so-called frailty models) were proposed to take into account the possible non independence of the observations. In this study, time until end-stage renal failure is a rightly censored end point. Possible intra-familial correlations due to shared environmental and/or genetic factors could induce dependence among familial failure times. In this paper, we fit marginal and frailty proportional hazards models to evaluate the effect of mutation type on the risk of IRT and an interfamilial heterogeneity of failure times. RESULTS: In this study, the use of these models allows to show the presence of an interfamilial heterogeneity of the failure times to IRT. Moreover, the results suggest that some mutation types are linked to a higher risk of fast evolution to IRT, which explains partially the interfamilial heterogeneity of the failure times. CONCLUSIONS: This paper shows the interest of marginal and frailty models to evaluate the heterogeneity of censored responses and to study relationships between a censored criterion and covariables. This study puts forward the importance of characterizing the mutation at a molecular level to understand the relationship between genotype and phenotype. PMID- 10673589 TI - [Analysis of a multicenter clinical trial on pressure ulcer development by a marginal approach in the Cox model]. AB - BACKGROUND: We exemplify the use of a marginal approach with proportional hazards model when failure times are correlated. METHODS: The marginal distribution for each failure time is formulated by the Cox proportional hazards model, while the dependence structure is unspecified. However, a correct variance-covariance estimate of the regression coefficients that takes into account the intra-group correlation is proposed. The program MULCOX2 which implements this statistical methodology is used to assess the effect of a nutritional supplementation intervention on pressure ulcer development on critically ill older patients from a multicentric trial (involving 19 wards). We compare the results obtained with those of the usual Cox regression. RESULTS: The naive approach yields much smaller standard error estimates of the regression parameters than the robust approach. CONCLUSION: In our example, the results obtained with the marginal approach do not modify the conclusions: a nutritional supplementation intervention tends to decrease significantly the formation of pressure ulcers. However in other situations, ignoring the intra-cluster dependence could lead to invalid statistical inference. The variability of the estimated effects by MULCOX2 can be quite sensitive to the number of clusters in the sample and to the clusters size. PMID- 10673590 TI - [Value of multinomial model in epidemiology: application to the comparison of risk factors for severely and moderately preterm births]. AB - BACKGROUND: The multinomial logistic regression model is employed to model the relationship between an outcome variable with more than two categories and a set of covariates. This model is not widely used in epidemiology. We discuss the value of the multinomial model by comparing it with the binary logistic model, and we present a statistical comparison of odds ratios (OR) using the multinomial model. We studied the associations between obstetric history and very (< 33 weeks of amenorrhea) and moderate (33-36 weeks) preterm births. METHODS: Parameters (lnOR) of very and moderate preterm births, associated with the severity of obstetric history (none=0, moderate=1, severe=2), were estimated using two logistic binary models (moderate preterm births vs full-term births (>=37 weeks), and very preterm births vs full-term births) and one logistic multinomial model which compared very and moderate preterm births to full-term births. These analyses were performed before and after adjustment for a covariate: the country of survey. Parameters of very preterm birth and moderate preterm birth, estimated from multinomial model, were compared using Wald test. These analyses were performed using data from a large case-control survey in Europe, the EUROPOP survey; 1 675 very preterm births, 3 652 moderate preterm births and 7 965 full term births were included. RESULTS: Crude parameters of very and moderate preterm births were similar, regardless the logistic regression model, binary or multinomial. The estimated parameters slightly differ after adjustment for the covariate, but lower variance estimates were obtained using multinomial logistic regression model. Parameters of very preterm birth associated with moderate obstetric history, B(gp)=0.5040, and severe obstetric history, B(gp)'=1.545, differ significantly from those of moderate preterm birth, B(pm)=0.4434 and B(pm)'=1.223 respectively (p < 0.001). CONCLUSION: Parameters obtained in separate logistic binary models are close to those obtained in a multinomial model. The multinomial model is useful for testing the heterogeneity of risk factors for distinct health problems. PMID- 10673591 TI - [Internal statistical validation of a quality of life questionnaire]. AB - In this paper we present two different statistical approaches to evaluate the psychometric properties of a quality of life questionnaire. First the study of the factorial structure is briefly exposed. Then we present the unidimensional classical models. They are based on the linear relationship between the observed score and the true score. The definition of the reliability was first addressed in this classical framework. Its estimation with the Cronbach alpha coefficient is one important feature of the evaluation of an instrument. More recently, modern response theory gives a better statistical framework to deal with unidimensional latent traits. These models describe the probability of positive answer to an item as a function of the actual value of the latent trait and an item parameter. We expose the principles of the Rasch model: hypothesis, estimations methods and fit tests. Finally practical applications to the validation process of a questionnaire are explored with data from a study of a short French version of the SIP questionnaire. PMID- 10673592 TI - [Use of GEE for modeling censored correlated data: application to the study of risk factors for withdrawal of totally implantable vascular access devices in cystic fibrosis]. AB - BACKGROUND: The proportional hazards model proposed by Cox for modeling censored data is not suited for correlated delays, for instance when several events can be observed on each subject. METHODS: To analyze correlated delays, we propose to use a log-linear marginal model equivalent to Cox model. Correlations are taken into account through the use of Liang and Zeger's Generalized Estimating Equations (GEE) and of their robust variance estimator. An advantage of this method is that it can be implemented through the SAS GENMOD procedure. When ties are observed, we propose to use multiple imputations, creating M data sets without ties from the original one. RESULTS: This method is applied to a retrospective survey on the risk of withdrawing totally implantable vascular access devices (TIVAD) because of complication in cystic fibrosis patients: 265 TIVAD implanted in 200 patients were observed. Risk factors were characteristics of the device or of the patient. Results obtained with the robust variance estimator and ten imputations show that the use of the device for taking blood (vs exclusive perfusion of antibiotics), polyurethane catheter (vs. silicon), use of counterpressure for upkeeping and pulmonary colonization by Pseudomonas Aeruginosa are significantly associated to withdrawal. Under the Cox model which does not account for the correlations, some conclusions differ because the robust variance of the estimators is smaller than the variance obtained under the working assumption of independent delays. CONCLUSION: This approach allows the modeling of correlated survival data with SAS software. Our results illustrate the necessity of accounting for existing correlations. PMID- 10673593 TI - [Logistic regression vs other generalized linear models to estimate prevalence rate ratios]. AB - In cross-sectional studies, to quantify the association between a risk factor and a disease (possibly adjusted for confounders), in the framework of the multiplicative model, the more obvious effect measure is a prevalence rate ratio with an associated confidence interval. The validity of this confidence interval requires an unbiased estimator and an appropriate estimate of the variance. In numerous epidemiological studies however, routine use is made of odds ratios and logistic regression. As the odds ratio per se is difficult to understand, prevalence odds ratios are often interpreted as prevalence rate ratios. But this latter approximation is valid only under the rare disease assumption. Moreover, in the logistic regression model, the variance of the estimates is based on the assumption of binomial variability, which is not always supported by the data; in the frequent case of overdispersion, this leads to under-estimation of the type I error rate. Yet, within the generalized linear model, it is easy to choose a link function other than the logit. For example, the log link (log-binomial model) is appropriate to directly estimate adjusted prevalence rate ratios. In case of overdispersion, it is also possible to achieve a better fit of the model, either by choosing another distribution in the exponential family or by estimating a dispersion parameter for the binomial distribution. Thus, there are no valid reasons for the systematic choice of odds ratio and of the logistic regression model to estimate prevalence rate ratios, unless the type of study imperatively requires their use. PMID- 10673594 TI - [Competitive risks and multi-state models in epidemiology]. AB - An introduction to multi-state models is presented. Survival models can be considered as simple multi-state models; models with competing risks allow to consider several mortality causes: they enable estimating cause-specific mortality rates; however, care must be exerted for the interpretation of survival functions in such models. The illness-death model is very useful for the study of chronic diseases. Transition intensities can be estimated using conventional Cox models when observations are done in continuous time. In that model too, one must avoid pitfalls when interpreting survival functions. PMID- 10673595 TI - [Survival analysis example based on an event history model from a clinical trial in cardiology]. AB - BACKGROUND: The main purpose of this paper is to present a survival analysis example based on an event history model in a competitive risks framework. Our example is derived from a clinical trial designed for comparing survival of a beta-blocker therapy group and a placebo group. METHODS: Competitive risks under study were non lethal cardiovascular events, permanent treatment withdrawals and deaths. The event history model was assumed to be semi-Markovian. The Cox proportional hazards model was used for modeling effects of treatments and regression variables on the transition rates. RESULTS: After taking into account all covariables influencing survival, mortality in patients who had one non lethal cardiovascular event after randomization but remained treated was shown to be reduced in the beta-blocker therapy group (RR=0.41; CI 95%=[0.17-0.98], p=0.04). CONCLUSIONS: Our analysis indicates that beta-blocker therapy should not necessarily be stopped in patients who experience a non lethal cardiovascular event under treatment. This finding requires confirmation in a separate setting. PMID- 10673596 TI - [Prognostic factors of recurrence and/or death in colorectal cancer: multistate modeling]. AB - Analysis of survival of patients with cancer sets particular epidemiological and statistical problems, especially when one wants to take into account metastasis or local recurrences. Cox's model does not allow modeling multiple events. Wei et al. have proposed an extension of Cox's model, by formulating the marginal distributions of multivariate failure times, which allows testing covariates effects on different events. We applied these methods to data from the Registry of Digestive Tumors of Burgundy, France. Prognostic factors of recurrence are rectal location of tumor and advanced stage at diagnosis. Prognostic factors of death are male gender, age greater than 75, rectal location and advanced stages. However, relative risk of recurrence for advanced stages is significantly greater than relative risk of death (p=4.10(-3)), while tumor location has the same influence on the two events. PMID- 10673597 TI - [Using capture-recapture models to estimate transition rates between states in interval-censored data]. AB - We introduce capture-recapture models, survival models developed in animal population biology to estimate survival / transition rates between sites within a population. We then show how to use these models in epidemiology or clinical research, to estimate survival / transition rates between stages of a disease (e.g. cancer, AIDS...), when patients enter a longitudinal study with imperfect follow-up resulting in interval-censoring. This method yields unbiased estimates, since the compliance (defined here as the probability of visiting the doctor at the planned date) is modelled jointly. We apply the method to cancer data, and show that "time" (elapsed since entry of a patient in the study) affects compliance in these data. PMID- 10673599 TI - [In Process Citation] PMID- 10673598 TI - [In Process Citation] PMID- 10673600 TI - 2000 ! les ABC, volume 58, numero 1 PMID- 10673601 TI - [An update on tools]. PMID- 10673602 TI - [SFBC and European clinical biology. Societe Francaise de Biologie Clinique]. PMID- 10673604 TI - [Representation of SFBC]. PMID- 10673603 TI - [SFBC and Inserm]. PMID- 10673605 TI - [Fifty years of biology with SFBC]. PMID- 10673606 TI - [Assets and liabilities of SFBC]. PMID- 10673607 TI - [Promoting the teaching of clinical biology]. PMID- 10673608 TI - [Biomedicine for the sake of the patient]. PMID- 10673609 TI - [The bright future of biology]. PMID- 10673610 TI - [Concerning national quality control of glycosylated hemoglobin (HBA1C)]. PMID- 10673611 TI - [Some historic milestones of clinical haematology]. AB - Microscopy, cell staining methods and tests for characterization of coagulation factor deficiencies are the landmarks of hematology at the beginning. Direct access to bone marrow, automatization, development of in vitro culture systems, monoclonal antibodies, biochemistry of proteins and nucleic acids were used to build today conception of the blood. Definitions of stem cells, growth factors, chromosomal translocations in leukemias and coagulation cascade represent the various aspects of the complex scientific object that the blood became. PMID- 10673612 TI - [DNA chips]. AB - DNA chips represent a miniaturization of the classical system of reverse dot blot. They consist of a small size support made of plastic or glass or silicium on which probes are synthesized or immobilized. It is thus possible to fix a few thousand or even a hundred of thousands of probes per cm2. Practically the chips are hybridized with the nucleic acid to be studied that has been amplified beforehand by PCR and which is generally labelled by a fluorochrome, either during amplification or after hybridization. After washing the hybrids are detected by a system which most of the time consists of a laser and a confocal microscope interfaced with a computer, but many variations exist. It is thus possible to analyse at the same time a considerable number of sequences. The diagnostic applications are only at the prototype stage. Eventually the chips should allow the identification of any point mutation, or the search for bacteria, viruses or parasites in a very short period of time without preliminary cultures. They should also allow many sorts of typing ranging from infectious agents to HLA. They should become a particularly powerful tool in the search for new medicines and in the revealing of their toxicity. A considerable potential market is also the diagnosis of the predisposition to polygenic diseases or to a particular sensitivity to any chemical substance. In fact the chips are only just beginning to be used. One of the foreseeable developments should be the possibility to study nucleic acids without preliminary amplification and we can hope to eventually have at our disposal very cheap, autonomous integrated systems. The technology could eventually extend to fields other than molecular biology such as immunology or biochemistry. PMID- 10673613 TI - [Troponins: biological and clinical aspects]. AB - The troponin complex is composed of three subunits called troponin I, troponin T and troponin C. Heart isoforms of troponin I and T are not expressed in skeletal muscle. They can be assayed in blood samples by immunoassays in order to diagnose and monitor patients with heart diseases. Troponins released in the blood after necrosis are mainly found as binary or ternary complexes. The kinetics of their release are close to that of creatine-kinase MB, but the return to normal is observed after longer periods of time. The excellent analytical sensibility of troponin assays allows the detection of minor heart damages. Because of its high specificity, the heart isoform of troponin I is use to diagnose heart damages in the presence of damages to other organs such as muscle and kidney. The resynthesis of troponin T by the skeletal muscle decreases its interest to diagnose muscle-associated heart diseases. In spite of the present lack of standardisation of troponin I assays, its detection associated to an other early myocardial marker, such as myoglobin, allows a rapid and reliable diagnostic of myocardial infarction. The study of post-translationally modified forms of troponins and their assays could permit a diagnosis and a more precise follow-up of patients with cardiac necrosis. PMID- 10673614 TI - [Procalcitonin, a new marker for bacterial infections]. AB - Procalcitonin (PCT), the precursor protein of the hormone calcitonin, appears to be an early marker of the presence of severe systemic infection. High serum concentrations are associated with severe systemic bacterial, parasitic or fungal infections. In contrast, PCT is generally not induced by severe viral infections or inflammatory reactions of non-infectious origin. Hence, PCT can be used for differential diagnosis of bacterial and viral meningitis. PCT may be helpful in the differentiation between infectious and non-infectious origin of systemic inflammatory response syndrome (SIRS) and acute respiratory distress syndrome (ARDS), pancreatitis, cardiogenic shock and acute rejection of organ transplants. PCT monitoring may be useful in patients with high risk of bacterial infection (major surgery, trauma, immunocompromised patients). PCT is a very stable molecule in vitro, and its measurement requires only 20 ml of plasma or serum and can be done within 2 hours. PMID- 10673615 TI - [Strategy for serum ferritin measurement in 16 to 45 year old women in health screening centers]. AB - Even in industrialized countries, the iron-deficiency anemia is frequent in menstruating women. However, the systematic measurement of serum ferritin is not justified. In this study, a strategy for ferritin measurement has been determined from data of centers for health screening, obtained in 6,098 menstruating women. This strategy is based on biological results (hemoglobin, MCV, RDW, GGT, ALAT) and on responses to the questions about blood donation, birth country and contraceptive habits. The measurement of serum ferritin is realized in 64% menstruating women and 23% have an hypoferritinemia (< 20 mg/l). PMID- 10673616 TI - [A new technique for measuring 17beta-estradiol using Kryptor (Cis Bio International): utilization to monitor ovulation stimulation]. AB - We studied the performances of the 17b-estradiol determination with the Kryptor System (Cis Bio international), in assisted medical procreation cycles. The detection is based on Trace technology. We compared this method with Estradiol-6 method on ACS-180 (Bayer Diagnostics). The Kryptor method is sensitive, has a good precision and accuracy. Comparison with ACS-180 showed a good correlation but results were much higher. Patients under stimulation for assisted procreation were followed by the two methods. The results were close to those obtained with ACS-180 but significantly higher. Thus it was necessary to define the levels which have to be reached before ovulation induction. PMID- 10673617 TI - [Helicobacter pylori: identification and detection of clarithromycin resistance by gene amplification]. PMID- 10673618 TI - [Focus on a new assay of fucosylated alpha-fetoprotein and evaluation in the biological diagnosis of hepatocellular carcinoma in cirrhosis]. PMID- 10673619 TI - [Platelets satellitism with polynuclear neutrophils]. PMID- 10673621 TI - [Investigation of isolated aspartate aminotransferase elevation in a patient treated with minocycline: characterization of a macro-enzyme]. PMID- 10673620 TI - [Infectious mononucleosis or sleeping sickness?]. PMID- 10673622 TI - [Thrombotic microangiopathy revealing acute monoblastic leukemia (LAM5)]. PMID- 10673623 TI - [Quality assurance: internal quality control and external quality evaluation]. PMID- 10673624 TI - [Rules concerning the transport of diagnostic specimens by road. Working Group of the SFBC. Quality proceeding of the Quality Control section of the SFBC]. PMID- 10673625 TI - Editorial PMID- 10673626 TI - [TP53 tumor suppressor gene: 20 years (and ten thousand mutations) later]. PMID- 10673627 TI - [Karyotype, aging, and cancer types: are they linked?]. PMID- 10673628 TI - [Cytogenetics of malignant lymphomas in the year 2000]. PMID- 10673630 TI - [Evolution of cancer markers: from radio-immunology to DNA chips and from surveillance to forecasting]. PMID- 10673629 TI - [Role of the ATM gene in genetic predisposition to cancer]. PMID- 10673631 TI - [Anatomy and pathologic cytology in cancerology: yesterday, today, and tomorrow]. PMID- 10673632 TI - [Therapeutic update on cancer of the breast]. PMID- 10673633 TI - [High-dose chemotherapy in ovarian adenocarcinoma]. PMID- 10673634 TI - [Update on the medical treatment of urologic tumors]. PMID- 10673635 TI - [Laparoscopic surgery in gynecologic oncology in 2000]. PMID- 10673636 TI - [Pharmacokinetic monitoring]. PMID- 10673637 TI - [Preventing and treating hematologic complications of anticancer chemotherapy: update on hematopoietic growth factors]. PMID- 10673638 TI - [Cancer immunotherapy: new approaches]. PMID- 10673639 TI - [Antitumor cellular immunotherapy: the breakthrough of dendritic cells]. PMID- 10673640 TI - [Quality of life in cancerology]. PMID- 10673641 TI - Stand there, don't just do something. PMID- 10673643 TI - Osseointegration or osteointegration? PMID- 10673644 TI - Can academics do it all? PMID- 10673645 TI - Destructive midline palatal lesion. PMID- 10673646 TI - Leukemia. PMID- 10673647 TI - Hydroxyapatite cement for calvarial reconstruction. PMID- 10673648 TI - Is the mandibular third molar a risk factor for mandibular angle fracture? AB - OBJECTIVES: The objective of this study was to evaluate the association of mandibular angle fractures with the presence and state of the eruption of the mandibular third molar (M3). METHODS: The medical records and panoramic radiographs of 615 patients with mandibular fractures were examined. The presence or absence and degree of impaction of the M3 were assessed for each patient and related to the occurrence of fracture of the mandibular angle. Data were also collected for age, sex, mechanism of injury, number, and location of mandibular fracture. Data were analyzed by a chi-square statistic test and Student t test. RESULTS: The incidence of mandibular angle fracture was found to be significantly greater when an unerupted M3 was present (P <.05). Of the 426 patients with an M3, 127 (29.8%) had angle fractures. Of the 189 patients without an M3, 25 (13.2%) had angle fractures. CONCLUSIONS: The results of this study showed that the mandibular angle that contains an impacted M3 is more susceptible to fracture when exposed to an impact than an angle without an M3. PMID- 10673649 TI - Prospective study of modified condylotomy for treatment of nonreducing disk displacement. AB - OBJECTIVE: This study was performed to provide an objective assessment of the outcome of modified condylotomy for treatment of the painful temporomandibular joint with nonreducing disk displacement (Wilkes late stage III, IV, V). STUDY DESIGN: A prospective study of 31 consecutive patients (43 joints) was conducted. All patients had nonreducing disk displacement verified by means of disk imaging. Independent evaluations were performed to assess pain, dysfunction, and progression of disease. The examinations were performed before modified condylotomy and at intervals up to 1 year after the operation. Eighteen patients (26 joints) completed the required examinations. Patient-based assessments were completed for pain and diet on 15 of these 18 patients (23 joints) 3 years after the operation. RESULTS: Visual analog scale (VAS) scores (mean +/- SE) for pain improved from 7.4 +/- 0.4 before modified condylotomy to 2.4 +/- 0.5 1 year later (P <. 001). Joints with degenerative joint disease (Wilkes stage IV, V) had less satisfactory pain relief compared with stage III joints (3. 6 +/- 0.9 vs 1.1 +/- 0.4, P =.05) and an 11-fold higher risk (P <. 04) for serious residual pain (VAS score >4). Dietary restrictions improved from a mean VAS score of 5.3 +/- 0.7 before the operation to 7.7 +/- 0.5 1 year later (P =.02). Minor differences between mean VAS scores at 1 (2.1 +/- 0.5) and 3 (2.1 +/- 0.5) years for pain, and 1 (7.4 +/- 0.6) and 3 (8.1 +/- 0.6) years for diet, were not significant. Mean maximal interincisal opening was 36.7 +/- 2.0 mm before the operation, and this improved to 40.1 +/- 2.0 mm 1 year later (P <.02). Mean contralateral movement was 8.3 +/- 0.5 mm before the operation and 8.4 +/- 0.6 mm 1 year after the operation (P >.05). None of the 12 Wilkes late III joints progressed to Wilkes IV or V, and none of the 14 Wilkes IV, V joints showed evidence of further bone resorption. The rate for reoperation was 4%. Minor complications occurred in 5 patients and were resolved in all but 1 a year later. When these outcomes were judged by 7 American Association of Oral and Maxillofacial Surgeons assessment indices for internal derangement, the mean rate of favorable outcome was 87%. CONCLUSION: Modified condylotomy is a safe and effective operation for treating pain and diminished function of temporomandibular joints with nonreducing disk displacement. It also seems to be an effective treatment for slowing further progression of the internal derangement and associated pathologic conditions. PMID- 10673650 TI - Submandibular gland mucocele: diagnosis and management. AB - Mucoceles originating from the submandibular gland are extremely rare. A review of the English literature resulted in identification of only 5 such cases. We have diagnosed and treated 2 submandibular mucoceles. Both lesions were removed in continuity with the submandibular and sublingual glands. No complications and no recurrences have occurred to date. The diagnosis of these lesions is complicated because of the lack of specific clinical diagnostic criteria and the similarity between submandibular mucoceles and plunging or cervical ranulas. Computerized tomography and specifically the presence of a so-called "tail" sign is pathognomonic for plunging ranula. This sign is absent in mucoceles originating in the submandibular glands. The treatment strategies vary as well. A diagnostic algorithm and a surgical rationale for treatment of submandibular mucoceles are presented. PMID- 10673651 TI - Efficacy of high-density versus semipermeable PTFE membranes in an elderly experimental model. AB - OBJECTIVE: The clinical effectiveness of expanded polytetrafluoroethylene (PTFE) membranes for guided bone regeneration has been reported in the literature on several occasions. However, one major drawback of this material is the need for a second surgical operation for removal of the membrane. In addition, most studies involving guided bone regeneration in animal models have used young animals or animals of unspecified age, but in some clinical situations guided bone regeneration may be indicated in elderly jaw bones. The objectives of this investigation were (1) to evaluate the effectiveness of a newly introduced high density PTFE membrane (TefGen-FD)-a material that does not require second-stage surgery for its removal-for enhancement of bone ingrowth in elderly rabbits' calvaria and (2) to compare the findings with those obtained with the commonly used semipermeable expanded PTFE membrane (Gore-Tex). STUDY DESIGN: Eighteen elderly rabbits (each more than 30 months old) served as the experimental animals in this study. Two non-self-healing, full-thickness defects were created in each rabbit calvarium. One of the 2 defects was fully covered with macroprous expanded PTFE membrane (Gore-Tex); the other defect was covered with microporous PTFE membrane (TefGen-FD). Specimens were obtained at 4, 8, and 16 weeks and examined by means of light microscopy. RESULTS: Clinically, the high-density TefGen membrane was much easier to detach from the underlying bone than was the semipermeable Gore-Tex membrane, which showed strict adherence to bone surface on removal. Microscopically, the Gore-Tex membrane lamellae were infiltrated by fibro-osseous tissue at the membrane's lower surface. A relatively greater speed and quantity of bone regeneration were observed in the defective cavities covered with Gore-Tex membrane than in those covered with TefGen membrane. CONCLUSIONS: It appears that the semipermeable Gore-Tex membrane is more effective than the high-density TefGen-FD membrane with respect to guided bone regeneration in elderly bone. These findings have relevance for the clinical situation of using guided bone regeneration in conjunction with implant placement and ridge augmentation procedures in atrophic elderly jaws. PMID- 10673652 TI - Oral staphylococcal mucositis: A new clinical entity in orofacial granulomatosis and Crohn's disease. AB - OBJECTIVE: Orofacial granulomatosis and the oral manifestations of Crohn's disease comprise many clinical features, of which stomatitis is one. The purpose of this study was to establish a role for Staphylococcus aureus in mucositis affecting some patients with orofacial granulomatosis or oral Crohn's disease. STUDY DESIGN: Four patients (2 with orofacial granulomatosis and 2 with oral Crohn's disease), from a total of 450 patients examined over 10 years, had stomatitis involving the entire oral mucosa, from which S aureus was cultured by the oral rinse technique. These patients were treated with flucloxacillin or erythromycin. RESULTS: A heavy growth of S aureus was isolated from the mouth of each patient. All 4 patients responded to treatment with flucloxacillin or erythromycin. CONCLUSIONS: S aureus is a potential cause of panstomatitis in patients with orofacial granulomatosis or Crohn's disease. This infection responds rapidly to antimicrobial treatment. PMID- 10673653 TI - Delayed eruption of permanent teeth in hyperimmunoglobulinemia E recurrent infection syndrome. AB - OBJECTIVE: To determine the incidence of abnormal tooth eruption in patients with hyperimmunoglobulinemia E (hyper-IgE) syndrome. STUDY DESIGN: This study evaluated 34 individuals with hyper-IgE syndrome (age range, 2-40 years). A comprehensive dental history and a head and neck evaluation were performed on all patients. Dental age was assessed in patients younger than 17 years by 2 methods: (1) clinical assessment of tooth eruption and (2) a radiographic method. Relationships between the chronologic age, dental developmental age, and age at tooth eruption were determined. Other oral or dental anomalies were recorded. RESULTS: Of patients older than 7 years, 75% reported problems with permanent tooth eruption, as evidenced by retained primary teeth or the need for elective extractions of primary teeth to allow eruption of permanent teeth. None of the patients experienced problems with eruption of primary teeth. Eruption of the first and second permanent molars also occurred on time. Dental maturity scores were established for 14 patients 17 years of age or younger. In each case, the difference between chronologic age and the estimated dental developmental age was less than 12 months; however, we found a significant discrepancy between the chronologic age and the mean age of tooth eruption in 80% of these patients when using a particular set of standardized values. Persistence of Hertwig's epithelial root sheath was observed on histologic examination. Chronic multifocal oral candidiasis was a consistent feature in patients with hyper-IgE recurrent infection syndrome. Other oral anomalies were also noted. CONCLUSION: We confirmed that a disorder of tooth eruption is part of the hyper-IgE syndrome. This problem occurs because of delayed primary tooth exfoliation rather than a developmental delay in the formation of the permanent dentition. The persistence of Hertwig's epithelial root sheath is unusual and may be associated with the lack of resorption of the primary teeth. Dentists should be aware of this feature of hyper-IgE syndrome because timely intervention will allow normal eruption to occur. PMID- 10673654 TI - Past administration of beta-lactam antibiotics and increase in the emergence of beta-lactamase-producing bacteria in patients with orofacial odontogenic infections. AB - OBJECTIVE: The purpose of this study was to determine the current status of beta lactamase-producing bacteria in orofacial odontogenic infections. STUDY DESIGN: Microbiologic data regarding purulent exudate from 111 cases with orofacial odontogenic infections were analyzed in relation to the past administration of beta-lactams. RESULTS: beta-lactamase-producing bacteria were isolated more frequently from the beta-lactam-administered group (38.5%) than from the beta lactam-nonadministered group (10.9%; P <.005), and they were isolated more frequently as the duration of administration increased. The predominant bacteria isolated included Prevotella (the most frequent isolate), viridans streptococci, Peptostreptococcus, and Fusobacterium, and 7.1% of total isolates produced beta lactamase. Penicillin and cefazolin worked well with beta-lactamase-nonproducing Prevotella but were remarkably affected by beta-lactamase-producing Prevotella. Cefmetazole, sulbactam/cefoperazone, and imipenem worked well against both types of Prevotella. CONCLUSIONS: beta-lactams are still suitable for the first antimicrobial therapy in the treatment of these infections. However, because past beta-lactam administration increases the emergence of beta-lactamase-producing bacteria, beta-lactamase-stable antibiotics should be prescribed to patients with unresolved infections who have received beta-lactams. PMID- 10673655 TI - Examination of the oral mucosa and peripheral blood cells of patients with recurrent aphthous ulceration for human herpesvirus DNA. AB - OBJECTIVE: The purpose of this study was to exam the oral mucosa and peripheral blood cells of patients with recurrent aph-thous ulceration (RAU) for the presence of the following human herpesviruses: herpes simplex viruses 1 and 2, varicella zoster virus, Epstein-Barr virus, cytomegalovirus, human herpesvirus-6, and human herpesvirus-7. STUDY DESIGN: Fifty-eight subjects with RAU and 10 control subjects were recruited at an academic referral center and enrolled in this prospective, nonrandomized, case-controlled study. Each of the subjects with RAU was seen during an acute episode, and swab specimens from lesional (RAU acute/lesion) and clinically normal (RAU-acute/normal) oral mucosa were obtained. Each of 2 subjects with RAU was evaluated during more than one acute episode. Three subjects with RAU were seen between active episodes, and swab specimens were taken from clinically normal (RAU-convalescent) oral mucosa. Swab specimens from clinically normal (control/normal) oral mucosa were obtained from the control subjects. Peripheral blood specimens were obtained from subjects with RAU and control subjects at the time the swab specimens were performed. Through use of polymerase chain reaction, all swab and peripheral blood specimens were examined for the presence of human herpesvirus DNA. Statistical significance was determined by means of chi(2) analysis. RESULTS: Herpes simplex virus and human herpesvirus-6 were found in a higher percentage of mucosal specimens from the control subjects (herpes simplex virus, 4/10; human herpesvirus-6, 5/9) than from the subjects with RAU (RAU-acute/lesion: 3/45 herpes simplex virus, 13/53 human herpesvirus-6; RAU-acute/normal: 7/48 herpes simplex virus, 9/53 human herpesvirus-6). No difference was demonstrated between RAU-acute/lesion, RAU acute/normal, and RAU-convalescent mucosal specimens for any of the human herpesviruses. Different human herpesviruses were identified from individual subjects with RAU during subsequent episodes of disease. Epstein-Barr virus (6/35), human herpesvirus-6 (3/40), and human herpesvirus-7 (7/43) were detected in the peripheral blood mononuclear cells during acute RAU but not in RAU convalescent or control peripheral blood mononuclear cells. CONCLUSIONS: The detection of human herpesvirus DNA from the oral mucosa and peripheral blood mononuclear cells of patients with RAU appears to represent normal viral shedding rather than a direct causal mechanism in this disorder. PMID- 10673656 TI - Calcified leiomyoma of the lateral pterygoid muscle in an 8-year-old boy. AB - Deep soft tissue leiomyomas are extremely rare benign tumors in childhood. An unusual case of benign calcified leiomyoma of the pterygoid muscle in an 8-year old boy is presented. Clinical manifestations and radiographic and histologicpathologic findings, as well as the differential diagnosis and possible histogenesis of this rare tumor, are discussed. PMID- 10673657 TI - Plasma cell granuloma of the oral mucosa with angiokeratomatous features: a possible analogue of cutaneous angioplasmocellular hyperplasia. AB - We report a plasma cell granuloma arising in the movable mucosa of the oral cavity of a 50-year-old man. Histologically, the lesion was characterized by a dense nodular infiltrate of mature plasma cells. Immunostaining for kappa and lambda light chains confirmed a polyclonal plasma cell population. Elongated rete ridges of the overlying epithelium formed collarettes around dilated blood and lymph vessels in focal areas. Based on the overall histologic architecture, we hypothesize that these peculiar changes are secondary to local blood flow alteration with congestive vasodilation caused by a dense plasma cell infiltrate. We believe that the plasma cell population may represent the oral counterpart of the cutaneous angioplasmocellular hyperplasia. PMID- 10673658 TI - Oral findings in DiGeorge syndrome: clinical features and histologic study of primary teeth. AB - OBJECTIVE: For the purpose of supplementing the shortage of dental information about DiGeorge syndrome, we report two cases of the syndrome seen in Japanese boys. STUDY DESIGN: Two cases were compared with respect to orofacial and dental findings; one was a case of complete DiGeorge syndrome and the other a case of partial DiGeorge syndrome. Extracted deciduous teeth from the two boys underwent histologic study. RESULTS: Each patient showed systemic developmental delay, hypocalcemia, and slight mental retardation. In the orofacial area, hypertelorism, a short philtrum, thick and reflected lips, and hypoplasia of the nasopharynx were also observed. A dental examination showed delayed formation and eruption of permanent teeth, aplasia of the nasopharynx, and enamel hypoplasia along with enamel hypocalcification. Structural streaks with increased calcification were histologically detected in the deciduous tooth from the patient with complete DiGeorge syndrome. CONCLUSIONS: Common characteristic orofacial and dental findings were noted in the two DiGeorge syndrome cases. Furthermore, histologic study of the deciduous tooth from the boy with complete DiGeorge syndrome suggests that there was some relationship between transient relative hypercalcemia and dentinal hypermineralized streaking of the tooth. PMID- 10673659 TI - Mercury release from dental amalgam after treatment with 10% carbamide peroxide in vitro. AB - OBJECTIVES: The effect of 10% carbamide peroxide on mercury release from dental amalgams was assessed in vitro by using a cold-vapor atomic absorption Mercury Analyzer System. STUDY DESIGN: Samples of 4 commercial brands of dental amalgam, Megaloy (Dentsply/Caulk, Milford, Del), Mega+ (CFPM, Aulnaye, France), Nongama 2 (Silmet, Or Yehuda, Israel), and Valiant Ph.D. (Dentsply/Caulk, Milford, Del), were treated for 48 hours with 10% carbamide peroxide and compared with samples treated with phosphate buffer. RESULTS: Amalgam specimens exposed for 48 hours to 10% carbamide peroxide showed significantly higher concentrations of mercury in solution as compared with specimens treated with phosphate buffer (P <.001). Megaloy and Valiant Ph.D. yielded significantly higher mercury concentrations in solution than Mega+ and Nongama 2 (P <.001). Mega+ yielded significantly higher mercury concentrations in solution than Nongama 2 (P <.05). No significant differences were found in mercury concentrations in solution between Megaloy and Valiant Ph.D. CONCLUSIONS: Treatment with 10% carbamide peroxide bleaching agents caused an increase in mercury release from amalgam restorations, possibly increasing exposure of patients to its adverse effects. Amalgam brands differed in the amounts of mercury release after bleaching with carbamide peroxide. PMID- 10673660 TI - Management of a horizontal root fracture after previous root canal therapy. AB - This case report concerns a 9-year-old girl who had a horizontal root fracture of her maxillary left central incisor and had root canal therapy performed by her family dentist immediately after the injury. Because of the incomplete canal obturation, the root canal filling material was removed, and calcium hydroxide therapy was initiated. Healing at the fracture site occurred, with hard tissue forming between the root segments. The coronal segment of the root was then obturated with gutta-percha. The patient later had orthodontic treatment with some movement of the coronal segment. Six-year post-treatment follow-up shows apparent clinical and radiographic success. This case illustrates the potential for repair of a horizontal root fracture after endodontic retreatment of the coronal segment and long-term splinting. PMID- 10673661 TI - A light and transmission microscopic study of mechanically exposed monkey pulps: dynamics of fiber elements during early dentin bridge formation. AB - OBJECTIVE: The aim of this investigation was to assess the role of the von Korff like fibers (VKF) during the process of dentin bridging. STUDY DESIGN: The monkey pulps, exposed to a hard-set calcium hydroxide, were evaluated at 3, 7, 14, 21, 30, 90, and 180 days. RESULTS: At 21 days, longitudinal and transverse collagen fibrils were organized as lamellar structures in close proximity to and subjacent to the newly formed predentin. VKF bundles were present between newly formed odontoblastoid cells. The VKF were bundles of thin collagen fibrils inserted into the predentin, which consisted of thick collagen fibrils. At 30 days, the exposure site was completely occluded with a new predentin matrix; lamellar structures and VKF were no longer present. CONCLUSIONS: The VKF may play a role in the connective tissue attachment to the dentin bridge, early in its formation. PMID- 10673662 TI - Pulpal tissue in bilateral talon cusps of primary central incisors: report of a case. AB - Talon cusp is a tooth anomaly that affects both the primary and the permanent dentitions. However, the occurrence of this anomalous cusp is rather infrequent in the primary dentition. Only 7 cases of bilateral talon cusps affecting the primary teeth have been reported in the dental literature. This is a case report of bilateral talon cusps on primary maxillary central incisors whose histologic evaluation revealed the existence of pulpal tissue in the anomalous cusps. PMID- 10673663 TI - Effective dose from radiation absorbed during a panoramic examination with a new generation machine. AB - OBJECTIVES: The purpose of this investigation was to measure the tissue-absorbed dose and to calculate the whole-body effective dose (E) for a new generation panoramic machine (Planmeca PM 2002 CC Proline), operating in the panoramic examination mode. Comparisons could then be made with historical panoramic and intraoral radiographic dose measurement values and effective dose estimates. STUDY DESIGN: Thermoluminescent dosimeters were embedded in a humanoid, tissue equivalent phantom at anatomically significant sites, representing key tissues. Absorbed dose measurements were obtained after every 5 panoramic exposures of a 25-exposure total. The measured average tissue-absorbed doses from a single panoramic exposure were used in the calculation of the whole-body E. RESULTS: The whole-body E for the PM 2002 CC Proline panoramic examination is 3.85 microSv. This is below the panoramic average of 6.7 microSv. The PM 2002 CC Proline panoramic examination delivers approximately 5% to 12% of the E of a complete mouth intraoral radiographic examination. CONCLUSIONS: The effective dose from the PM 2002 CC Proline examination is at the low end of the range for other panoramic machines and is far below either a D-speed or E-speed film intraoral radiographic examination. PMID- 10673664 TI - Mandibular bone density and fractal dimension in rabbits with induced osteoporosis. AB - OBJECTIVE: Our goal in this investigation was to examine the mandibular bone density and radiographic textural changes and the relationship between mandibular and spinal bone mineral density in an osteoporotic rabbit model. STUDY DESIGN: Three adult female New Zealand white rabbits in each of 4 groups received daily injections of cortisone acetate at a dosage of 0.0 (control), 3.0, 7.5, or 15.0 mg/kg for 4 weeks. The rabbits were then killed, and the mandible and spine of each animal were removed. Digital radiographs (70 kVp, 10 mA, 8 impulses) of the hemimandibles and spines were made. Lateral and anteroposterior bone densities of the lumbar spine (L2) were calculated, and average mandibular interdental bone density, fractal dimension, and gradient values were calculated. RESULTS: Correlation analysis revealed that cumulative steroid dose was strongly related to mandibular bone density (r = -0.80, P <.01), moderately related to mandibular fractal dimension (r = -0.61, P <. 05), and moderately related to anteroposterior lumbar spine density (r = -0.64, P <.05). Moderate correlations were found between mandibular interdental bone density and spinal density (r = 0.56, P <.05), but mandibular fractal dimension was not related to spinal density. CONCLUSIONS: In osteoporotic female rabbits, mandibular bone mineral density decreases in relation to spinal density and cumulative steroid dose. Mandibular fractal dimension decreases with cumulative steroid dose but is not significantly related to either mandibular density or spinal density. PMID- 10673665 TI - Radiographic manifestation of clear cell odontogenic tumor. AB - Clear cell odontogenic tumors are uncommon, only 20 cases having been reported in the literature. We report a case that presented with unique radiographic manifestations over a period of 12 years; no similar case has been reported to date. Documentation of the malignant nature of the condition and of the range of clinical and radiographic manifestations with which this neoplasm can present provides useful insight into its pathogenesis and progression. Early and timely recognition of the lesion, histopathologic examination, and aggressive interventional procedures are in order to successfully treat this condition and prevent a potentially fatal outcome. PMID- 10673666 TI - Radiographic evidence of enamel pearls in jordanian dental patients. AB - OBJECTIVE: An "enamel pearl" is an ectopic globule of enamel that is adherent to the tooth root surface. Such an anomaly may facilitate the progression of periodontal breakdown. Information on the prevalence of enamel pearls is sparse, and ethnic variations are thought to occur. Our objective was to assess the prevalence of enamel pearls in a group of Jordanian dental patients. STUDY DESIGN: A random sample of 819 dental records were selected, and a total of 1032 periapical radiographs were interpreted for the presence of enamel pearls. RESULTS: Enamel pearls were detected in 4.76% of the subjects and on 1.6% of the molars. No significant difference between sexes was observed. First molar teeth were the most commonly affected, whereas the third molars were the least affected with the condition. CONCLUSIONS: Enamel pearls are not uncommon among Jordanian dental patients, and their early detection could be important in prevention of periodontal disease. PMID- 10673667 TI - Ganglioneuroma of the mandible: radiologic and pathologic findings of a rare tumor. AB - Ganglioneuroma is a rare benign neural tumor, thought to arise from the ganglia of the sympathetic chain or parasympathetic nervous system. It is therefore most commonly seen in the abdomen and thorax. The occurrence of ganglioneuroma in the mandible is exceptional; to date only 5 cases have been reported. We present another case of ganglioneuroma of the mandible, demonstrate the radiologic and pathologic findings, and discuss the possible causes of the tumor at this site. PMID- 10673668 TI - Dose-intense paclitaxel, etoposide and cyclophosphamide: a safe and active regimen for tumor cytoreduction and stem cell mobilization in metastatic breast cancer. AB - Patients with metastatic breast cancer in complete remission are the ones most likely to have an improved outcome with subsequent high-dose chemotherapy and autologous peripheral blood stem cell transplantation (HDC-PBSCT). Peripheral blood stem cells are usually procured following mobilization with single agent chemotherapy and colony-stimulating factor support. We utilized a dose-intense regimen of paclitaxel 200 mg/m2 i.v., etoposide 60 mg/kg i.v., and cyclophosphamide 3 g/m2 i.v. (TEC) followed by daily administration of granulocyte colony-stimulating factor. The aim was not only to mobilize stem cells but also to achieve optimal tumor cytoreduction prior to HDC/PBSCT. One hundred consecutive patients with metastatic breast cancer received 257 cycles of TEC between March 1994 and June 1997, with the aim of collecting 5 x 106 CD34 positive cells/kg usually following the second cycle of chemotherapy. Patient characteristics included a median age of 45 years, a median of two organ systems involved by disease, a median of two prior chemotherapy regimens and eight prior chemotherapy cycles, and a median interval of 8 months from diagnosis of metastases to first cycle of TEC. There were 61 febrile episodes during neutropenia and 13 of these were associated with bacteremia or fungemia. Mortality rate was 1%. An adequate number of stem cells was collected in 90% of patients. The overall response rate of the tumor was 58.8% with 23.7% complete responders among 97 evaluable patients. Multivariate analysis demonstrated chemosensitivity to the most recent standard chemotherapy regimen administered for metastatic disease, an ECOG performance score of 0 as opposed to 1, 2 or 3, and involvement by disease of only one organ system as significant variables for achieving a complete remission with TEC. This novel dose-intense regimen was safe and well tolerated, highly active against metastatic breast cancer, and capable of excellent stem cell mobilization. Bone Marrow Transplantation (2000) 25, 123 130. PMID- 10673669 TI - Hematopoietic stem cell allografts using a non-myeloablative conditioning regimen can be safely performed on an outpatient basis: report of four cases. AB - Using a non-myeloablative, immunosuppressive, fludarabine-based conditioning regimen, we performed allogeneic peripheral blood stem cell transplants totally on an outpatient basis in four patients (two with chronic myelogenous leukemia, one with acute myelogenous leukemia and one with thalassemia major). The median granulocyte recovery time to 0.5 x 109/l was 10 days and the lowest absolute neutrophil count was 0.064 x 109/l; only one patient developed thrombocytopenia below 20 x 109/l. No patient required red blood cell transfusions and one was given a single prophylactic platelet transfusion. All patients are alive at 210 390 (median 285) days and have definite evidence of chimerism; one developed biopsy-proven GVHD on day 50, with a limited cutaneous rash. The procedure is less costly than its counterpart using myeloablative conditioning regimens and may represent another approach in the management of patients requiring an allogeneic stem cell transplant. Bone Marrow Transplantation (2000) 25, 131-133. PMID- 10673670 TI - Evidence of alloreactive T lymphocytes in fetal liver: implications for fetal hematopoietic stem cell transplantation. AB - The use of hematopoietic stem cells for in utero transplantation to create permanent hematochimerism represents a new concept in fetal therapy, although this approach has provided heterogeneous results. In this paper we have undertaken molecular, phenotypic and functional studies aimed at identifying the presence of fully competent T lymphocytes in samples of fetal livers and cord blood. We found mature VDJ TCR beta chain transcripts in fetal liver cells taken from 7 to 16 weeks of gestation and a similar pattern was detected in cord blood cells sampled from 13.5 to 20.5 weeks of gestation. A Vbeta8 gene sequence comparable to that detected in adult PBMC was found in fetal liver samples at 9 or 17 weeks gestation. PreTalpha message was detected in all samples and its expression decreased in fetal blood samples with increasing gestational age while Calpha message appeared at 9.4 weeks and its expression increased during gestational age. T cell clones obtained from fetal liver cells showed a mature TCR alphabeta+, CD8+ phenotype and displayed strong alloreactivity against allo MHC class I molecules. The presence of alloreactive T lymphocytes may explain the failure to engraft in fetuses older than 13 to 16 weeks and may provide insights into fetal liver transplantation. Bone Marrow Transplantation (2000) 25, 135-141. PMID- 10673671 TI - Cytogenetic status pre-transplant as a predictor of outcome post bone marrow transplantation for chronic myelogenous leukaemia. AB - We have analysed pre-transplant cytogenetic findings in 418 patients with CML in pre-blastic phase who underwent allogeneic BMT between February 1981 and January 1998. Five different patient groups were identified: A = Philadelphia (Ph)+; B = Ph-, BCR-ABL+; C = variant Ph (VPh); D = Ph chromosome plus at least one of: trisomy 8, +Ph, chromosome 17 abnormalities and E = other abnormalities in addition to the Ph chromosome. There were two principal conclusions. Firstly, Ph- patients showed a better outcome, and VPh patients a worse outcome, than those with a standard Ph, both in terms of leukaemia-free survival (LFS) (76.9%, 22.1% and 31.9%) and the risk of treatment failure relative to those with a standard Ph (relative risks of 0.49 and 1.92, respectively). One contributing factor may be relapse: no Ph- patients relapsed, whereas all other groups showed similar probabilities of relapse at 5 years (range 33.0-44. 0%). Secondly, those with the additional changes of +8, +Ph and i(17q) did not show a worse outcome than those with no additional changes (5 year survival of 44.7% vs 51.8%; 5 year LFS of 40.6% vs 31.9%), whereas those with other additional changes may fare worst of all (40.4% and 16.0%, respectively). Bone Marrow Transplantation (2000) 25, 143 146. PMID- 10673672 TI - Incidence and outcome of vancomycin-resistant enterococcal bacteremia following autologous peripheral blood stem cell transplantation. AB - A retrospective evaluation of 321 consecutive recipients of high-dose chemotherapy (HDC) and autologous peripheral blood stem cell transplantation (PBSCT) was conducted to ascertain the incidence and outcome of vancomycin resistant enterococcal (VRE) bacteremia. Ten patients developed VRE bacteremia at a median of 6 days following PBSCT. Nine isolates were Enterococcus faecium and one was E. faecalis. The median duration of bacteremia was 5 days. The central venous catheter was removed in seven individuals. Nine patients were treated with a variety of antimicrobial agents including quinupristin-dalfopristin, chloramphenicol, doxycycline, oral bacitracin, co-trimoxazole, and nitrofurantoin. Bacteremia resolved without adverse sequelae in seven patients. Two individuals who died of other causes had persistent or relapsed bacteremia at the time of death. An additional patient suffered multiple relapses of VRE bacteremia and died as a result of VRE endocarditis 605 days following PBSCT. Mortality as a direct result of VRE bacteremia was 10% in this series. The optimal type and duration of treatment of VRE bacteremia has not been clearly defined. Therefore, we perform weekly stool surveillance cultures for VRE in our hospitalized transplant population and apply strict barrier precautions in those individuals in whom stool colonization has been identified. Furthermore, the empiric use of vancomycin has been restricted. Bone Marrow Transplantation (2000) 25, 147-152. PMID- 10673673 TI - Human herpesvirus 8 (KSHV) contamination of peripheral blood and autograft products from multiple myeloma patients. AB - Human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), has recently been identified within the bone marrow dendritic cells of multiple myeloma (MM) patients. This virus contains homologues to human cytokines such as IL-6 that could potentially stimulate myeloma cell growth and contribute to disease pathogenesis. Since mobilization chemotherapy may increase circulating dendritic cell numbers, we searched for HHV-8 in peripheral blood mononuclear cells (PBMCs) before and after mobilization chemotherapy given to MM patients. Furthermore, we determined if autograft purging using the CEPRATE SC device would reduce the percentage of HHV-8 infected stem cell products. Only two of the 39 PBMC samples collected prior to mobilization chemotherapy contained PCR detectable virus, yet nine of 37 PBMCs collected on the first day of leukapheresis had detectable HHV-8 (P = 0.016). HHV-8 was more frequently identified in autograft products before vs after Ceprate SC selection (40% vs 15%, P = 0.016). Although the role HHV-8 plays in myeloma pathogenesis remains unclear, these results imply that mobilization chemotherapy increases the numbers of circulating HHV-8-infected dendritic cells within the peripheral blood. In addition, CD34 selection of autograft products in MM patients may reduce the reintroduction of virally infected cells following high-dose chemotherapy. Bone Marrow Transplantation (2000) 25, 153-160. PMID- 10673674 TI - Respiratory syncytial virus immune globulin treatment of lower respiratory tract infection in pediatric patients undergoing bone marrow transplantation - a compassionate use experience. AB - Respiratory syncytial virus (RSV) pneumonia in BMT recipients carries a mortality rate of approximately 50-70% despite ribavirin (Virazole) treatment. In both immunocompetent and immunocompromised animal models, RSV neutralizing antibodies rapidly reduce pulmonary virus load after a single dose. RSV-IGIV (RespiGam) is an IgG immune globulin with high concentrations of RSV neutralizing antibody (>19 200 MU/ml). From June 1991 to February 1996, a compassionate-use protocol using RSV-IGIV for treatment of RSV infections was conducted. Eleven children at multiple centers, mean age 3.3 years (4 months to 9 years), were undergoing BMT and met the protocol criteria. They received a single 1500 mg/kg dose of RSV-IGIV infused over 12 h at a median of 5 days (1-37 days) after RSV symptom onset. Ten of these patients received prior or concurrent aerosolized ribavirin. Serum RSV neutralizing titers were measured in five patients and showed a 3- to 30-fold increase 24 h after RSV-IGIV infusion. Adverse events were mild. One of 11 (9.1%) patients died from their RSV illness (91% RSV survival). In comparison to previously published reports, RSV-IGIV treatment of RSV pneumonia in BMT patients may increase survival above that in such patients treated with ribavirin alone. Bone Marrow Transplantation (2000) 25, 161-165. PMID- 10673675 TI - Incidence, risk factors and outcome of varicella-zoster virus infection in children after haematopoietic stem cell transplantation. AB - We report a retrospective analysis of VZV infection after haematopoietic stem cell transplantation (HSCT) in children. Thirty-three (30%) of the total 109 children who were transplanted during a 7 year period developed post-transplant VZV infection. Twenty-four of these 33 (73%) children had VZV infection within 1 year following HSCT. The cumulative incidences of post-transplant VZV infection at 1 and 5 years were 26% and 45%, respectively. The positive and negative predictive values of pretransplant VZV serology in recipients on the development of HZ following HSCT were 39% and 88%, respectively. Pretransplant VZV seropositivity in recipients was the only risk factor for post-transplant herpes zoster (HZ) infection on multivariate analysis. All patients responded to acyclovir. The median duration of VZV infection was 5 days. Three (11%) and one (3%) children with HZ developed visceral dissemination and post-herpetic neuralgia, respectively. No mortality was directly attributed to VZV infection. VZV infection remains a major cause of morbidity in children after HSCT. Further studies are warranted to evaluate the potential use of VZV vaccine in these children. Bone Marrow Transplantation (2000) 25, 167-172. PMID- 10673676 TI - Evaluation of acute toxicities associated with autologous peripheral blood progenitor cell reinfusion in patients undergoing high-dose chemotherapy. AB - Peripheral blood progenitor cell reinfusion (PBPC) in patients undergoing high dose chemotherapy (HDC) for poor prognosis malignancies, has been described as causing possible acute gastrointestinal (nausea, vomiting), allergic (oedema, bronchospasm, anaphyl- axis), renal (proteinuria, haematuria) and/or cardiovascular (hypotension, arrhythmia, conduction disturbances, transient ischaemic phenomena) toxicities. To establish the clinical relevance of these observations and the possible relationship with different HDC regimens used, we performed a clinical and instrumental evaluation on 33 patients with advanced breast cancer, non-Hodgkin's lymphoma, Hodgkin's disease, relapsed ovarian cancer, Ewing's sarcoma, extragonadal germinal tumour and small cell lung cancer. They underwent at least one reinfusion each for a total of 51 studied procedures. No patient had a previous history of cardiovascular disease or significant intercurrent illness such as diabetes or liver, renal or neurologic impairment. All patients had totally implanted central venous catheters, through which the transplants had been collected and reinfused without technical consequences. To evaluate cardiovascular function, we continuously monitored 12-lead ECGs, with arterial pressure (AP) measurements every 5 min from the beginning of the procedure to 15 min after the reinfusion ended. We did not observe any significant differences between basal and subsequent steps in AP, heart rate, PQ and QTc time, P wave and QRS complex duration or P wave and QRS electrical axes. No patient showed any ST-T tract pathological abnormality, but one patient developed a transient ectopic atrial rhythm, without any haemodynamic disfunction and with spontaneous reversion to sinus rhythm. No patient complained of symptoms of haemodynamic failure. Gastrointestinal side-effects appeared to be strictly related to speed of reinfusion and to the number of packs reinfused, probably reflecting on the amount of dimethylsulphoxide infused. In one patient a tonic clonic seizure occurred during a vomiting episode, but no patient developed allergic or renal toxicities. We conclude that PBPC reinfusion, if managed according to the procedure we propose in patients without organic impairment, is a safe procedure not associated either with increased risk of acute arrhythmias or ischaemic or significant systemic acute toxicities. Bone Marrow Transplantation (2000) 25, 173-177. PMID- 10673677 TI - Anti-A isoagglutinin as a risk factor for the development of pure red cell aplasia after major ABO-incompatible allogeneic bone marrow transplantation. AB - Delayed erythropoiesis and pure red cell aplasia (PRCA) have been reported after major ABO-incompatible BMT. We attempted to find risk factors for the development of PRCA in 27 patients who underwent major ABO-incompatible BMT. In all patients, the donor marrow was depleted of RBCs before infusion. In 22 patients, isoagglutinins were determined until they disappeared. In eight (29.6%) out of 27 patients, bone marrow examination following BMT showed the findings of PRCA. We analyzed various clinico-pathologic risk factors and isoagglutinin type was the only significant risk factor. Patients with anti-A isoagglutinins against donor RBC developed PRCA more frequently than patients with anti-B (8/17 vs 0/9). Median days to the disappearance of isoagglutinins tended to be longer in patients with PRCA (PRCA vsnon-PRCA, 200 vs 66 days) and in cases with anti-A isoagglutinins (anti-A vsanti-B, 160 vs 51 days). Times to disappearance of isoagglutinins correlated with times to reticulocytes over 1% and initial appearance of donor type RBC (R2 = 0.708 and 0.711). In conclusion, RBC engraftment following major ABO-incompatible BMT was dependent on the disappearance of isoagglutinins against donor RBC, and anti-A isoagglutinin was a risk factor for the development of PRCA after major ABO-incompatible allogeneic BMT. Bone Marrow Transplantation (2000) 25, 179-184. PMID- 10673678 TI - Cardiac conduction abnormalities in patients with breast cancer undergoing high dose chemotherapy and stem cell transplantation. AB - Cardiac toxicities in 39 consecutive patients with breast cancer receiving high dose chemotherapy (HDC) with stem cell transplantation were reviewed. All 39 patients received various anthracycline-containing regimens in adjuvant settings and/or for metastatic disease before HDC. As a cytoreductive regimen, all received cyclophosphamide 2000 mg/m2 and thiotepa 200 mg/m2 for 3 consecutive days. No immediate fatal toxicities were observed, but one patient developed chronic congestive heart failure and two had transient left ventricular dysfunction. Pericardial effusion was observed in another three patients. ST-T abnormalities during HDC were observed in two patients and arrhythmias were observed in nine, four of which occurred during stem cell infusion (SCI). There were three atrial arrhythmias, two ventricular arrhythmias, and four atrioventricular (AV)-block episodes. Two patients developed advanced and complete AV-block with an asystolic pause. Notably, three patients experienced AV block with uncontrolled vomiting. No relationship was observed between the cumulative dose of anthracycline and cardiac toxicities during HDC. These results suggest that abnormalities in the conduction system during HDC may be more frequent than previously reported. Vagal reflex secondary to emesis may play an important role in the development of AV-block. Bone Marrow Transplantation (2000) 25, 185-189. PMID- 10673679 TI - Bone mass after allogeneic BMT for childhood leukaemia or lymphoma. AB - The bone mass was measured by dual energy X-ray absorptiometry in 25 survivors of childhood leukaemia or lymphoma (21 with ALL) who had received TBI and allogeneic BMT a median of 8 years ago (range 4-13). Results were compared with local data on 463 healthy controls and 95 survivors of childhood ALL treated without BMT. Adjusted for sex and age, the mean whole-body bone mineral content (BMC) and bone mineral areal density were significantly less than in healthy controls (0.8 and 0.5 s.d. less than predicted). The reduced BMC was caused by a significantly reduced height for age, whereas bone area for height and BMC for bone area were similar to controls. Less bone mass tended to be related to additional cranial irradiation and age above 20 years at follow-up. Controlled for this, the whole body bone mass seemed to be unrelated to previous chemotherapy and endocrine status at follow-up and tended to be only marginally less in BMT patients than in ALL survivors treated without BMT. In conclusion, 8 years after allogeneic BMT for childhood leukaemia or lymphoma, the whole-body bone mass was only slightly reduced and the size-adjusted bone mass (BMC for bone area) was normal. Bone Marrow Transplantation (2000) 25, 191-196. PMID- 10673680 TI - Evaluation of optimal survival of primitive progenitor cells (LTC-IC) from PBPC apheresis products after overnight storage. AB - Optimal overnight (ON) storage of PBPC aphereses is becoming an increasingly important issue and different options for storing PBPC products exist. The survival of primitive progenitor cells is of major interest, as recent data suggest that these progenitors are not only important for long-term engraftment but also contribute significantly to the early phase of hematopoietic engraftment after myeloablative therapy. We therefore investigated the survival of primitive progenitor cells (ie long-term culture initiating cells, LTC-IC) before (ie within 2 h after finishing the apheresis procedure) and after ON storage lasting 16 to 20 h. In addition, we compared the % of recovery of LTC-IC with that of mature progenitors (ie colony-forming cells, CFC) and with the % viability of the mononuclear cells in the apheresis product. Aliquots of PBPC aphereses products were tested in collection bags at room temperature (RT), in EDTA tubes both at RT or 4 degrees C +/- the addition of autologous plasma (AP; 2.6-fold the apheresis volume) and +/- the possibility of gas exchange. Mean viable cell counts did not show strong differences between the different storage conditions and were poor predictors for the survival of CFC and LTC-IC. At RT (collection bags, EDTA tubes +/- gas exchange) recoveries (% of input) of both, CFC (18%, 18% and 31%) and LTC IC (10%, 4%, 17%) were low. The addition of AP at RT improved the survival of CFC and LTC-IC to 66% and 38%, respectively. Optimal recoveries for both types of progenitors (CFC: 99%, LTC-IC: 109%) were obtained at 4 degrees C in the presence of AP. In addition, a good correlation between the survival of CFC and LTC-IC was obtained (r = 0.76) suggesting that the analysis of CFC may also allow some conclusions to be drawn on the survival of LTC-IC. Bone Marrow Transplantation (2000) 25, 197-200. PMID- 10673681 TI - Busulfan clearance in renal failure and hemodialysis. AB - The impact of hemodialysis on the clearance of busulfan was determined in a patient with chronic renal failure undergoing autologous peripheral stem cell transplantation for non-Hodgkin's lymphoma. The extraction ratio for busulfan across the dialyzer was 0.530 +/- 0.026 at a blood flow of 400 ml/min, which corresponds to a hemodialysis clearance of 2.23 +/- 0.11 ml/min/kg body weight. Apparent oral clearance of busulfan without hemodialysis was 3.38 +/- 0.56 ml/min/kg. Thus, a 4 h hemodialysis session enhanced the apparent oral clearance of busulfan by 65%. We conclude that hemodialysis effectively removes busulfan from circulating blood, but a standard hemodialysis period (ie, 4 h) does not significantly alter busulfan exposure. Bone Marrow Transplantation(2000) 25, 201 203. PMID- 10673682 TI - Severe acute graft-versus-host disease occurring after syngeneic BMT for AML in a patient not given prior cyclosporin A therapy. AB - A syndrome akin to graft-versus-host disease in the recipient of syngeneic stem cells is hitherto described as being milder, self-limiting and confined to the skin. It is enhanced by prior cyclosporin A therapy. We describe here a recipient of a syngeneic marrow transplant who did not receive priming with cyclosporin A and yet developed severe and progressive graft-versus-host disease which necessitated and responded to high-dose immunosuppressive therapy. We believe that this is because the conditioning regimen in stem cell transplant acts to reset the immune system enabling it to recognise 'self' antigens. Bone Marrow Transplantation (2000) 25, 205-207. PMID- 10673683 TI - Epstein-Barr virus-associated lymphoproliferative disease after a cord blood transplant for Diamond-Blackfan anemia. AB - A 7-year-old boy with Diamond-Blackfan anemia (DBA) developed lymphoproliferative disease (LPD) after a cord blood transplant (CBT). 3.1 x 107/kg mononuclear cells from an HLA one-locus mismatched CB were transplanted after conditioning with total body irradiation (8 Gy), cyclophosphamide (200 mg/kg) and antithymocyte globulin (10 mg/kg). Complete engraftment occurred on day 33 post transplant. Despite the resolution of grade II graft-versus-host disease (GVHD), he died of lymphoma on day 130 post transplant. The tumor was of donor origin, indicating clonal proliferation of Epstein-Barr virus (EBV)-infected B cells. This is the first report of EBV-LPD after CBT. Post-transplant LPD can be a serious EBV associated complication of CB grafts. Bone Marrow Transplantation (2000) 25, 209 212. PMID- 10673684 TI - A successful cord blood transplant in a child with second accelerated phase chronic myeloid leukemia following lymphoid blast crisis. AB - We describe a 5-year-old girl with Ph(+) CML who received a cord blood transplant in a second accelerated phase after a very early lymphoid blast crisis. She was induced into CR by ALL-directed chemotherapy and then maintained with IFN-alpha2b together with weekly rotational chemotherapy. Nineteen months after diagnosis, her mother gave birth to an HLA-compatible sibling, whose cord blood was cryopreserved. The patient's second acceleration occurred 22 months after the CML diagnosis. The subsequent conditioning regimen included busulfan 16 mg/kg, Ara-C 12 g/m2 and melphalan 140 mg/m2. In order to prevent GVHD, CsA alone was administered, 3 mg/kg i.v. per day for a total of 40 days. The total number of nucleated cells infused was 0.8 x 108/kg, with CD34+ cells 1.8 x 106/kg and CFU GM 1 x 104/kg. Engraftment occurred on day +35. Respiratory distress, severe VOD and grade II acute gastrointestinal GVHD complicated the post-transplant period. No chronic GVHD occurred. The girl is alive 23 months after transplantation with complete donor chimerism; both Ph chromosome and bcr/abl RNA are negative. Bone Marrow Transplantation (2000) 25, 213-215. PMID- 10673685 TI - Allogeneic peripheral blood cell transplantation for hypereosinophilic syndrome with myelofibrosis. AB - Patients with hypereosinophilic syndrome (HES) display a very heterogeneous clinical picture ranging from asymptomatic cases to very aggressive forms. We report a 38-year-old woman with progressive HES who developed severe myelofibrosis and was treated by allogeneic stem cell transplantation, using peripheral blood (PBSCT) instead of bone marrow as the source of progenitor cells, after conditioning with cytoxan and busulphan. To the best of our knowledge, this is the first case of HES with myelofibrosis treated with PBSCT. The patient remains alive 8 months post-PBSCT, and bone marrow fibrosis has significantly decreased following transplantation. Bone Marrow Transplantation (2000) 25, 217-218. PMID- 10673686 TI - Successful treatment of relapsed CML after cord blood transplantation with donor leukocyte infusion IL-2 and IFNalpha. AB - A 3-year-old girl with BCR/ABL-positive CML relapsed after related HLA-identical cord blood transplantation. She was treated with three cycles of donor lymphocyte (DLI) infusion from her 15-month-old brother. Interferon alpha was added after the second DLI, whereas a trial of IL-2 had to be discontinued because of increasing immature myeloid cells in the blood smear. No signs of GVHD were observed, but she developed myelosuppression and needed one platelet and one red blood cell transfusion. She achieved a molecular remission after 6 months with transient molecular relapse followed by sustained remission for 15 months. Thus, DLI with or without interferon alpha might prove to be a promising treatment option with tolerable side-effects in relapsed CML after cord blood transplantation. Bone Marrow Transplantation (2000) 25, 219-222. PMID- 10673687 TI - Immunologic recovery of patients given CD34-selected peripheral blood progenitor cell transplantation for malignant diseases. PMID- 10673688 TI - Late-onset herpes simplex virus-associated interstitial pneumonia after allogeneic bone marrow transplantation. PMID- 10673689 TI - Extramedullary relapse in the so-called 'sanctuary' sites for chemotherapy after donor lymphocyte infusion. PMID- 10673690 TI - Establishing a BMT support group. PMID- 10673691 TI - Engraftment syndrome after autologous peripheral blood stem cell transplantation with high numbers of peripheral blood stem cells followed by granulocyte colony stimulating factor administration. PMID- 10673692 TI - Mobilization of hematopoietic progenitor cells with paclitaxel (taxol) as a single chemotheraupetic agent, associated with rhG-CSF. AB - We assessed the mobilization capacity of taxol with rhG-CSF, both as a single chemotherapeutic agent and in the presence of cyclophosphamide (CY), and compared the effect with yields achieved when mobilization was performed solely with rhG CSF. Fifteen patients with breast cancer received taxol 170 mg/m2 (continuous infusion, day 1) and rhG-CSF (8 microg/kg/day, from day 2 until the end of apheresis) (T-G group), while seven breast cancer patients were additionally treated with CY (4 g/m2) on day 2, followed by rhG-CSF starting at similar doses on day 3 (T-CY-G group). The PBSC collections after taxol with/without CY were compared with those of 30 breast cancer patients who had received rhG-CSF (8 microg/kg/day) for mobilization. No differences were found in the characteristics of patients included in any of the three mobilization groups. The median yield of CD34+ cells from all patients included in taxol containing schedules was 9 x 106/kg (range 2-26) collected with a median of one apheresis procedure (range 1 4). Leukaphereses began earlier in the T-G group (median day 8, range 7-10) than in the T-CY-G group (median day 13, range 11-17). In most patients (20 out of 22) who received taxol containing regimens, more than 2.5 x 106 CD34+ cells/kg, a threshold considered to be sufficient for hematopoietic reconstitution, were collected with a single apheresis. Those patients in the T-G group experienced less neutropenic and thrombocytopenic days, with all neutropenic fever episodes developing in patients treated with the T-CY-G schedule (43%). When considering priming with rhG-CSF alone in our historical cohort of 30 breast cancer patients, a significant detrimental effect was observed in comparison with taxol mobilizing schedules, in the number of aphereses performed, in the total yield CD34+cells and in the number of patients who achieved the target dose of 2.5 x 106/kg CD34+ cells within the first collection procedure. We conclude that taxol containing schedules are effective in mobilizing PBSC and facilitate the collection of high yields of CD34+ cells (usually more than 5 x 106/kg recipient body weight) with a reduced number of apheresis procedures. Taxol, as a single agent with rhG-CSF, exhibits less hematological toxicity than the combination chemotherapy mobilization regimen including CY. Bone Marrow Transplantation (2000) 25, 231 235. PMID- 10673693 TI - Prognostic significance of increased IL-10 production in patients prior to allogeneic bone marrow transplantation. AB - IL-10 is a potent immunosuppressant which inhibits allo-antigen-specific T cell responses. In addition, IL-10 is a strong endogenous anti-inflammatory cytokine. To investigate the role of IL-10 in the induction of acute GVHD following allogeneic bone marrow transplantation (BMT) we performed a prospective study on spontaneous IL-10 production by peripheral blood mononuclear cells (PBMNC) in 84 patients admitted for allogeneic BMT. High spontaneous IL-10 production by PBMNC at the time of admission and prior to any preparative treatment correlated with a subsequent low incidence of GVHD and transplant-related mortality (8%), as compared to patients with low or intermediate IL-10 production (50%, P < 0. 01). Our data demonstrate the prognostic significance of increased IL-10 production in BMT patients and suggest a major role of IL-10 in maintaining immunobalance in the setting of allogeneic BMT. Bone Marrow Transplantation (2000) 25, 237-241. PMID- 10673694 TI - Highly purified CD34+ cells isolated using magnetically activated cell selection provide rapid engraftment following high-dose chemotherapy in breast cancer patients. AB - The primary objective of this study was to evaluate the safety of infusion of CD34+ cells, selected using a clinical scale magnetically activated cell sorting device, assessed by time to hematological engraftment and incidence of adverse events. Secondary objectives included evaluation of device performance in terms of purity and recovery of the CD34+ cell product. Breast cancer patients suitable for transplantation received cyclophosphamide and filgrastim for mobilisation, followed by three leukaphereses. The products of the first two leukaphereses underwent CD34+ cell selection. The product of the third leukapheresis was cryopreserved unmanipulated. Following high-dose cyclophosphamide, thiotepa and carboplatin, selected CD34+ cells were infused. In 54 patients who received selected cells only, the median time to platelet recovery and neutrophil recovery was 11 days (range 5-51) and 9 days (range 5-51), respectively. There were no adverse events associated with infusion of selected cells. A total of 126 leukapheresis samples was available before and after selection for central CD34+ analysis. The median purity was 96.1% (27.4-99.4) and the median recovery was 52. 3% (15.2-146.3). These data show that cells selected using magnetically activated cell selection provide safe and rapid engraftment after high-dose therapy. Bone Marrow Transplantation (2000) 25, 243-249. PMID- 10673695 TI - Efficacy of autologous stem cell transplantation in mantle cell lymphoma: a 3 year follow-up study. AB - This study was designed to evaluate the efficacy of therapeutic intensification with autologous stem cell transplantation (ASCT) for mantle cell lymphomas (MCL) in terms of response rate, duration of response, and event-free and overall survivals. Twenty-four patients with confirmed MCL responding to chemotherapy received a high-dose chemo-radiotherapy regimen followed by ASCT. Transplantation was performed during first-line therapy in nine cases, second-line in 13 cases and third-line in two cases. The source of hematopoietic stem cells was peripheral blood for 19 cases. At the time of ASCT, eight patients were in complete remission (33%). Seventeen of the 24 cases received an intensified regimen with TBI and seven received the BEAM or the BEAC regimen. After transplantation, 19 patients were in CR (79%). Nine of these were alive in continued CR at a median follow-up of 34 months, while seven relapsed at a median of 18 months. One patient died from Pneumocystis carinii interstitial pneumonitis and five patients developed secondary malignancies. With a median follow-up after transplantation of 34 months, the 3-year event-free survival was 55% and the 3 year overall survival was 68%. These results indicate that therapeutic intensification with ASCT might be an effective treatment for mantle cell lymphomas. Bone Marrow Transplantation (2000) 25, 251-256. PMID- 10673696 TI - Intensified preparative regimens and autologous transplantation in refractory or relapsed intermediate grade non-Hodgkin's lymphoma. AB - Between September 1986 and June 1998, 99 patients with relapsed or refractory IGL received intensified preparative therapy and underwent autologous transplantation at a single institution. Two intensified preparative regimens were used: cyclophosphamide, etoposide, total body irradiation (CY-VP-TBI) (n = 66) and cyclophosphamide, BCNU, etoposide (CBV) (n = 33). As clinical features and results were not different for the two preparative regimens, results were combined. For all patients undergoing autologous transplantation, 5-year actuarial overall survival (OS) was 34% +/- 6%; 5-year event-free survival (EFS) was 26% +/- 5%. For patients who responded to primary therapy, salvage therapy, or both, OS was 42% +/- 7%; for non-responders to prior therapy, OS was 14% +/- 7%, P < 0.025. OS was better among patients responding to salvage therapy (50% +/ 9%), than among patients who had a complete response to initial therapy, but failed to respond or were untested/unevaluable with respect to salvage therapy (26% +/- 10%; P < 0.025). On multivariate analysis, response to salvage therapy was associated with survival following autologous transplantation (P < 0. 005). Treatment related mortality was 9% overall and only 6% after G-CSF and GM-CSF were introduced into routine clinical practice. High-intensity preparative therapy is highly effective, with acceptable treatment-related mortality, in patients with IGL who have responded to induction therapy, salvage therapy, or both. The best responses are observed in patients responding to salvage therapy. Randomized prospective studies will be needed to further define the role of intensified preparative regimens. Bone Marrow Transplantation (2000) 25, 257-262. PMID- 10673697 TI - The prophylactic potential of fludarabine monophosphate in graft-versus-host disease after bone marrow transplantation in murine models. AB - Fludarabine phosphate, a purine analogue currently used in the therapy of hematological malignancies, is known to cause immunosuppression and long-lasting T cell lymphopenia. In this study, the effect of fludarabine on murine graft versus-host disease occurring after marrow transplantation across major and minor histocompatibility barriers was evaluated. Survival of (BALB/c x C57BL/6)F1 mice irradiated and transplanted across the major histocompatibility barrier with C57BL/6 spleen cells, and subsequently treated with fludarabine was significantly longer than that of the control animals (P < 0.0001). On the other hand, fludarabine had no effect on the graft-versus-host disease and survival of CBA mice transplanted by B10.BR and of BALB/c mice transplanted by B10.D2 spleen cells across the minor histocompatability barrier. The results indicate that in certain murine models, particularly a major mismatch, fludarabine has the potential to induce bilateral tolerance and stable chimerism after marrow transplantation. Bone Marrow Transplantation (2000) 25, 263-266. PMID- 10673698 TI - Reconstitution of lymphocyte subpopulations after paediatric bone marrow transplantation. AB - We prospectively studied the reconstitution of lymphocyte subpopulations in a group of 22 children, who survived disease-free at least 6 months after allogeneic BMT for a haematological malignancy. Absolute counts of total lymphocytes, B lymphocytes, T lymphocytes, and CD4+ helper T lymphocytes reached the 5th percentile (p5) of age-matched reference values within 6 months after BMT in 15, 17, 7 and 2 patients, respectively. In particular, CD4+ helper T lymphocyte reconstitution was very slow. Unexpectedly, CMV reactivation had a profound positive influence upon the number of CD4+ helper T lymphocytes in the children. In five patients, absolute B lymphocyte counts above the 95th percentile were reached from 6 months after BMT onwards, mimicking normal ontogeny. Unlike normal ontogeny, the percentages of helper T lymphocytes expressing the 'naive' CD45RA isoform were low and those expressing the 'memory' CD45RO isoform were high in the first 3 months after BMT, as described before. Thereafter, the CD45RA:CD45RO ratio slowly normalised. Also, CD7 expression was absent on up to 90% of T lymphocytes in the first months after BMT, and on a steadily decreasing percentage thereafter, as recently described in adults. However, the absolute counts of CD45RO+/CD4+ and CD7-/CD4+ helper T lymphocytes did not change significantly. So, we found no evidence of peripheral expansion of previously primed donor-derived 'memory' T lymphocytes during the follow-up period which spanned 1-18 months after BMT. The absolute counts of 'naive' CD45RA+ helper T lymphocytes did not show a faster increase after BMT than in adults, despite the presumed presence of a non-involuted thymus in children. Bone Marrow Transplantation (2000) 25, 267-275. PMID- 10673699 TI - Isolation of viruses from stools in stem cell transplant recipients: a prospective surveillance study. AB - We prospectively examined stool specimens for enteric viruses in 75 stem cell transplant recipients (autologous 48, allogeneic 27) to determine the frequency and significance of these infections. Only six patients (8%) had a positive isolate. Five of these were allograft recipients (18%) compared to one autograft recipient (2%) (P = 0.02). Unrelated donor BMT recipients were at the highest risk for a viral isolate (OR = 10.5). Adenovirus was the commonest isolate (four patients). One patient each had an echovirus, enterovirus and small round structured virus identified. No correlation was found between the severity of gastro-intestinal symptoms and detection of a viral pathogen. There was no correlation with GVHD or CMV status. The only risk factor identified for isolation of an enterovirus was allogeneic BMT from an unrelated donor. There was a negative correlation with PBSC grafts. All the patients infected with an enteric virus had concomitant infection with other pathogens, compared to only 18% of uninfected patients (P = 0.001). The non-relapse mortality of the infected patients was 50% and only 7% in the uninfected patients (P = 0.01, OR = 12.5), although the isolated virus was the direct cause of death in one patient only. This study indicates a low rate of enteric virus isolation in recipients of PBSC grafts, both autologous and allogeneic. However, unrelated donor BMT is associated with a higher risk of enteric virus infection and an adverse outcome. Bone Marrow Transplantation (2000) 25, 277-282. PMID- 10673700 TI - A randomized placebo-controlled trial of lisofylline in HLA-identical, sibling donor, allogeneic bone marrow transplant recipients. The Lisofylline Marrow Transplant Study Group. AB - The purpose of the study was to evaluate the effect of lisofylline (LSF) on engraftment, regimen-related toxicities (RRT), and mortality in patients undergoing allogeneic bone marrow transplantation (BMT). We performed a multicenter, randomized placebo-controlled trial in 60 patients with hematologic malignancies receiving BMT from HLA-identical sibling donors. Patients were randomized to receive either placebo, 2 mg/kg LSF or 3 mg/kg LSF every 6 h, beginning before conditioning and continuing to day 21 or hospital discharge. Treatment groups were balanced with respect to conditioning regimen and disease stage. However, significantly more patients in the 2 mg/kg LSF group were at high risk for RRT due to performance status >/=1, age >/=40 years, and prior exposure to CMV. Nausea and vomiting were the only adverse events observed in a higher proportion of LSF-treated patients that led to study withdrawal in six of 42 patients (14%). The times to neutrophil recovery to >/=500/microl and platelet recovery (>20 000/microl) were not improved by LSF treatment. Nevertheless, no patient who received treatment with 3 mg/kg LSF developed a documented infection between day 0 and 35 or had a serious or fatal infection between day 0 and 100 (P = 0.003 vs placebo for both). The day-100 survival rate was also significantly improved in the 3 mg/kg LSF group (89%), compared with either the 2 mg/kg LSF (48%) or placebo (61%) groups (log-rank test, 3 mg/kg LSF vs placebo, P = 0. 026). We conclude that treatment with LSF 3 mg/kg reduced the incidence of infections and improved 100-day survival in patients receiving related-donor allogeneic bone marrow transplantation. Bone Marrow Transplantation (2000) 25, 283-291. PMID- 10673701 TI - Pulmonary cytolytic thrombi: a newly recognized complication of stem cell transplantation. AB - Over the past 5 years we have recognized a new pulmonary complication of hematopoietic stem cell transplantation (HSCT) associated with fever and pulmonary nodules termed 'pulmonary cytolytic thrombi' (PCT). Retrospective analysis of medical and radiographic records and pathologic material from 13 HSCT recipients with PCT and a review of the Blood and Marrow Transplant Database for all patients with radiographic evidence of pulmonary nodules or who underwent open-lung biopsy from 1 January 1993 to 31 December 1998 (n = 1228) were performed. The median age of patients with PCT was 11.9 years (range, 1.3-29.7 years). All patients developed fever at a median of 72 days (range, 8-343 days) post transplant, followed by pulmonary nodules on chest CT. Eleven patients were receiving therapy for active GVHD (acute, grades I-IV (n = 10); extensive chronic (n = 1)). Biopsy of the pulmonary nodules revealed a unique pattern of necrotic, basophilic thromboemboli with amorphous material suggestive of cellular breakdown products. This was descriptively labeled 'pulmonary cytolytic thrombi'. Immunohistochemical staining revealed entrapped leukocytes and disrupted endothelium, but was negative for histiocytes. Cultures and immunohistochemical stains were negative for infectious agents. Empiric therapy included systemic corticosteroids (n = 9) and amphotericin (n = 7). Nine patients survive with resolution of PCT at a median follow-up of 1.5 years. Bone Marrow Transplantation (2000) 25, 293-300. PMID- 10673702 TI - Neuropathological findings after bone marrow transplantation: an autopsy study of 180 cases. AB - We prospectively evaluated the neuropathological complications of 180 patients who underwent autopsy studies following bone marrow transplantation (BMT) (177 allogeneic, three autologous). The most frequent underlying disorders included severe aplastic anemia (n = 55), chronic myelogenous leukemia (n = 53), acute myelogenous leukemia (n = 24) and Fanconi anemia (n = 16). There were 114 males and 66 females. Neuropathological findings were detected in 90.55% of the patients. The most frequent findings were subarachnoid hemorrhages (SAH) (n = 57), intraparenchymal hemorrhages (IHP) (n = 49), fungal infections (n = 16), Wernicke's encephalopathy (n = 10), microglial nodular encephalopathy (n = 10) and neurotoxoplasmosis (n = 8). In only 17 patients was the brain within normal limits. Survival time after BMT averaged 5.4 months and the majority of patients died in the first 3 months post BMT (n = 105). Central nervous system (CNS) pathology was the main cause of death in 17% of the patients (n = 31), with a predominance of IHP in this particular group. Furthermore, the survival time of these patients who died of CNS causes (96.3 days) was almost half of the survival time of those who died of extra-cerebral causes (177.8 days) (P = 0.0162). IHP (70. 96 vs27.22%) (P < 0.001), fungal infections (25.8 vs 8.88%) (P < 0. 001) and toxoplasmosis (9.67 vs 4.44%) (P < 0.001) were significantly more frequent in the group of patients who died due to CNS causes than in the control group. The findings of this work provide a possible guide to the possible causes of neurological syndromes following BMT. Bone Marrow Transplantation (2000) 25, 301 307. PMID- 10673703 TI - Pulmonary toxicity following carmustine-based preparative regimens and autologous peripheral blood progenitor cell transplantation in hematological malignancies. AB - Sixty-five patients with hematological malignancies (25 multiple myeloma, 18 Hodgkin's disease, 22 non-Hodgkin's lymphomas) who received a carmustine-based regimen followed by autologous PBPC transplantation, were studied retrospectively to evaluate the incidence of post-transplant non-infective pulmonary complications (NIPCs), risk factors predictive of NIPCs, and response to steroids. Carmustine (BCNU) given i.v. at a dose of 600 mg/m2 was combined with etoposide and cyclophosphamide in 40 patients (BCV regimen) and with etoposide and melphalan in 25 patients (BEM regimen). Seventeen of 65 patients (26%) had one episode of NIPCs. The median time to NIPCs was 90 days (52-289). Factors that increased the risk of developing NIPCs on multivariate analysis were female sex (P < 0. 001) and BCV regimen (P < 0.05). All patients with NIPCs received prednisone at a dose of 1 mg/kg body weight for 10 days then tapered by 5 mg every two days; complete response to steroids was achieved in 15 of 17 patients; one unresponsive patient died of interstitial pneumonia. BCNU given at the dose of 600 mg/m2 is well tolerated when associated with melphalan and etoposide. In females and in patients receiving BCNU with cyclophosphamide, a BCNU dose reduction may be advisable. Bone Marrow Transplantation (2000) 25, 309-313. PMID- 10673704 TI - Mental disturbances during isolation in bone marrow transplant patients with leukemia. AB - The mental status of 39 leukemia patients, who received bone marrow transplants (BMT), was studied during the period of isolation. Mental disorders (diagnosed according to DSM-IV criteria) occurred in 16 patients (41%) during the observation period. The most frequent diagnoses were adjustment disorders, with anxiety and/or depression. Logistic regression analysis suggested higher Tension Anxiety score in the Profile of Mood States (POMS) prior to isolation (P = 0.011), donation of the bone marrow from unrelated subjects (P = 0.026) and in female patients (P = 0.033). The results are preliminary, but indicate a high frequency of mental disturbances and highlight the importance of psychiatric intervention in BMT patients. Bone Marrow Transplantation (2000) 25, 315-318. PMID- 10673705 TI - Sequential vidarabine infusion in the treatment of polyoma virus-associated acute haemorrhagic cystitis late after allogeneic bone marrow transplantation. AB - Late onset haemorrhagic cystitis (HC) occurs in 20-30% of allogeneic bone marrow transplant patients. Human polyomavirus BK (BKV) (or less frequently adenovirus) may be involved in the pathogenesis of viral HC and can represent a serious post transplant complication. Diagnosis and treatment of viral HC can be difficult and has an uncertain outcome. We report the efficacy of sequential vidarabine in the treatment of a patient with severe BKV-associated HC, despite the delay in implementing therapy. Bone Marrow Transplantation (2000) 25, 319-320. PMID- 10673706 TI - Typhlitis complicating autologous blood stem cell transplantation for breast cancer. AB - Three cases of typhlitis occurring during autologous blood stem cell transplantation (ABSCT) for metastatic breast cancer are described. Typhlitis is a rare complication of neutropenia and has uncommonly been reported in the autologous transplant setting. Although it has been most commonly described in children with leukemia, typhlitis has increasingly been reported in adult leukemias and in association with neutropenia secondary to chemotherapy for a number of solid tumors. Only five previous cases of typhlitis in the setting of ABSCT have been described. Whereas diarrhea and fever are common toxicities associated with high-dose chemotherapy, it is likely that many cases of typhlitis go unrecognized. Bone Marrow Transplantation (2000) 25, 321-326. PMID- 10673707 TI - Bone marrow transplantation in severe Glanzmann's thrombasthenia with antiplatelet alloimmunization. AB - Glanzmann's thrombasthenia is an autosomal recessive disorder characterized by a lack of platelet aggregation due to the absence of platelet glycoprotein IIb and IIIa. Usually, the disease leads to mild hemorrhage but sometimes bleeding is severe enough to be life-threatening. We report the case of a 16-year-old girl, presenting with very severe type 1 Glanzmann's thrombasthenia, successfully treated with an HLA-identical sibling bone marrow transplant (BMT). We also update the clinical and laboratory data of her brother, who had received a BMT 16 years ago for the same disease. In the light of these two cases and two others published in the literature, we discuss the indications for BMT from HLA identical sibling donors in Glanzmann's thrombasthenia. Alloimmunization against the missing platelet GPIIb/IIIa complex and severity of bleeding episodes may constitute sufficient criteria for allogeneic BMT after careful assessment of the risk-benefit of such a procedure, although this remains exceptional in this disease. Bone Marrow Transplantation (2000) 25, 327-330. PMID- 10673708 TI - FK506-induced intractable leukoencephalopathy following allogeneic bone marrow transplantation. AB - FK506-related leukoencephalopathy has been reported to be reversible and readily treated by discontinuation or reduction of FK506. We describe two pediatric cases of FK506-related leukoencephalopathy following allogeneic bone marrow transplantation, which could not be readily controlled. These cases show that FK506-related leukoencephalopathy is not always reversible, and patients may develop epilepsy. Bone Marrow Transplantation (2000) 25, 331-334. PMID- 10673709 TI - Mild pre-transplant cardiomyopathy may not impair long-term quality of life after bone marrow transplantation. AB - An 8-year-old child with acute myeloid leukemia (AML), underwent an allogeneic bone marrow transplant (BMT) from his HLA matched sister in spite of having a mild cardiomyopathy. We followed the patient with periodic electrocardiograms (ECG) and echocardiograms which have not worsened, and the patient's quality of life is not compromised 14 years after BMT. Bone Marrow Transplantation (2000) 25, 335-336. PMID- 10673710 TI - Syngeneic peripheral blood stem cell transplantation with brief immunosuppression for severe aplastic anemia. AB - We report the cases of two severe aplastic anemia (SAA) patients who were successfully treated with syngeneic peripheral blood stem cell transplantation (PBSCT) using immunosuppression without high-dose chemotherapy or irradiation for conditioning. A 21-year-old woman with SAA of 6 years duration had been transfused heavily before transplantation and had developed refractory thrombocytopenia, chronic hepatitis and secondary hematochromatosis. Syngeneic PBSCT with immunosuppression using ATG, methylprednisolone, and cyclosporin-A was eventually performed without high-dose chemotherapy in September 1997. The second syngeneic PBSCT with the same immunosuppression was successfully performed in a 35-year-old male patient who had had SAA for 3 months in November 1998. Haemopoietic engraftment was rapid and sustained. There was no infection or mucositis during the syngeneic PBSCT. The patients are currently 9 to 22 months post-PBSCT without rejection. Our experience suggests that syngeneic PBSCT with brief immunosuppression is an effective alternative to pretransplant high-dose chemotherapy conditioning for SAA patients having syngeneic transplantation. Bone Marrow Transplantation (2000) 25, 337-339. PMID- 10673712 TI - Response to Dr furebring PMID- 10673711 TI - Side-effects of amphotericin B lipid complex (Abelcet) in the Scandinavian population. PMID- 10673713 TI - The promise of nucleic acid vaccines. AB - Establishing the effective use of 'naked' nucleic acids as vaccines would undoubtedly be one of the most important advances in the history of vaccinology. While nucleic acids show much promise for use as vaccine vectors in experimental animals, not a single naked nucleic acid vector has been approved for use in humans. Indeed, data from human clinical trials is scant: nucleic acid vaccines have not been clearly demonstrated to have any convincing efficacy in the prevention or treatment of infectious disease or cancer. Here we illustrate possible mechanisms underlying effective nucleic acid vaccination. We focus on progress that has been made in the improvement of their function. Additionally, we identify promising new strategies and try to forecast future developments that could lead to the real success of nucleic acid vaccines in the prevention and treatment of human disease. PMID- 10673714 TI - Gene therapy in the CNS. AB - Gene therapy for neurological disorder is currently an experimental concept. The goals for clinical utilization are the relief of symptoms, slowing of disease progression, and correction of genetic abnormalities. Experimental studies are realizing these goals in the development of gene therapies in animal models. Discoveries of the molecular basis of neurological disease and advances in gene transfer systems have allowed focal and global delivery of therapeutic genes for a wide variety of CNS disorders. Limitations are still apparent, such as stability and regulation of transgene expression, and safety of both vector and expressed transgene. In addition, the brain adds several challenges not seen in peripheral gene therapy paradigms, such as post-mitotic cells, heterogeneity of cell types and circuits, and limited access. Moreover, it is likely that several modes of gene delivery will be necessary for successful gene therapies of the CNS. Collaborative efforts between clinicians and basic researchers will likely yield effective gene therapy in the CNS. PMID- 10673715 TI - Targeting adenovirus. AB - The use of targeted viral vectors to localize gene transfer to specific cell types holds many advantages over conventional, non-targeted vectors currently used in gene therapy. The resulting improvements in gene localization from targeted adenovirus vectors are likely to reduce immunogenicity and toxicity, increase safety, and enable the systemic administration of these vectors for multiple indications including cancer, cardiovascular disease, and inflammatory disease. Recent advances in the biological understanding of adenovirus structure and adenovirus receptor interactions have fueled the rapid development of targeted adenovirus vectors. Two basic requirements are necessary to create a targeted adenovirus vector: interaction of adenovirus with its native receptors must be removed and novel, tissue-specific ligands must be added to the virus. Two general approaches have been used to achieve these basic requirements. In the 'two-component' approach, a bispecific molecule is complexed with the adenovirus. The bispecific component simultaneously blocks native receptor binding and redirects virus binding to a tissue-specific receptor. In the 'one-component' approach the adenovirus is genetically modified to ablate native receptor interactions and a novel ligand is genetically incorporated into one of the adenovirus coat proteins. Two-component systems offer great flexibility in rapidly validating the feasibility of targeting via a particular receptor. One component systems offer the best advantages in producing a manufacturable therapeutic and in more completely ablating all native adenovirus receptor interactions. The coming challenges for targeted adenovirus vectors will be the demonstration that the technology performs in vivo. Ultimately, or in parallel, 'receptor-targeting' technology can be combined with improved adenovirus backbones and with 'transcriptional targeting' approaches to create adenovirus which deliver genes selectively, safely, and with little immune response. PMID- 10673716 TI - Design and application of HSV vectors for neuroprotection. AB - Herpes simplex virus has been extensively genetically modified for gene transfer to nerve and other tissues, to create vectors that are devoid of viral gene expression and toxicity. Recombinant vectors have been engineered to express genes which protect neurons against toxic insults resulting in cell death, including nerve growth factor (NGF) and anti-apoptotic genes (eg bcl-2). This review describes experiments using HSV vectors expressing these gene products and their potential protective role in ameliorating neurodegenerative processes in animal model systems. PMID- 10673717 TI - Regulated gene expression systems. AB - Most gene therapy research to date has focused on solving the delivery problem- getting genes efficiently and stably into target cells and tissues. Those working on systems for regulating the expression of genes once delivered have often been accused of trying to run before they can walk. Yet regulation is likely to be essential if gene therapy is to realize its full potential as a mainstream clinical option for delivering proteins. Dramatic progress has been made in designing and testing systems in which expression is controlled by orally active drugs. The next few years should see the first clinical trials of drug-regulated gene therapies. PMID- 10673718 TI - Evaluation of polyether-polyethyleneimine graft copolymers as gene transfer agents. AB - Cationic copolymers consisting of polycations linked to non-ionic polymers are evaluated as non-viral gene delivery systems. These copolymers are known to produce soluble complexes with DNA, but only a few studies have characterized the transfection activity of these complexes. This work reports the synthesis and characterization of a series of cationic copolymers obtained by grafting the polyethyleneimine (PEI) with non-ionic polyethers, poly (ethylene oxide) (PEO) or Pluronic 123 (P123). The PEO-PEI conjugates differ in the molecular mass of PEI (2 kDa and 25 kDa) and the degree of modification of PEI with PEO. All of these conjugates form complexes upon mixing with plasmids, which are stable in aqueous dispersion for several days. The sizes of the particles formed in these systems vary from 70 to 200 nm depending on the composition of the complex. However, transfection activity of these systems is much lower than that of PEI (25 kDa) or Superfect as assessed in in vitro transfection experiments utilizing a luciferase reporter expression in Cos-7 cells as a model system. In contrast, conjugate of P123 with PEI (2 kDa) mixed with free P123 (9:1(wt)) forms small and stable complexes with DNA (110 nm) that exhibit high transfection activity in vitro. Furthermore, gene expression is observed in spleen, heart, lungs and liver 24 h after i.v. injection of this complex in mice. Compared to 1,2-bis(oleoyloxy) (trimethylammonio) propane:cholesterol (DOTAP:Chol) and PEI (25 kDa) transfection systems, the P123-PEI system reveals a more uniform distribution of gene expression between these organs, allowing a significant improvement of gene expression in liver. PMID- 10673719 TI - Cell delivery, intracellular trafficking and expression of an integrin-mediated gene transfer vector in tracheal epithelial cells. AB - The mechanism of cell entry and intracellular fate of a gene transfer vector composed of a receptor-targeting, DNA-condensing peptide, RGD-oligolysine, a luciferase encoding plasmid DNA (pDNA) and a cationic liposome was examined. We demonstrate by confocal microscopy, electron microscopy and subcellular fractionation that the major mechanism of entry of the vector is endocytic. The vector complex rapidly (5 min) internalizes into early endosomes, then late endosomes and lysosomes. Entry involves, at least in part, clathrin-coated pit mediated endocytosis since different conditions or drugs known to influence this pathway modify both uptake of pDNA and its expression. The observed increase in expression with addition of a lip some correlated with an increase in the rate of transfer of the pDNA to lysosomes, a decrease in intracellular recycling and exocytosis of the pDNA and an increase in the amount of pDNA in the nuclear fraction. Trafficking within the cell involved endosome fusion and the acid environment of the endosomes-lysosomes was beneficial for expression. After 30 min both the peptide and pDNA localized to the nucleus and the amount of intact pDNA in the nuclear fraction was highest with liposome and peptide. A better understanding of the cellular mechanisms by which vectors transfer to and traffic in cells should help design improved vectors. PMID- 10673720 TI - Variegation of retroviral vector gene expression in myeloid cells. AB - We have comparatively evaluated the efficiency of a series of retroviral vectors transducing the gp91-phox gene, whose defects are responsible for impaired production of superoxide anion (O2-) by phagocytic cells and lead to the X-linked form of chronic granulomatous disease (X-CGD). These vectors included four constructs based on the MoMuLV backbone and expressing gp91-phox from the viral long terminal repeat (LTR) or from internal promoters, and one construct based on the myelotropic FMEV vector. Expression of the therapeutic gene from the MoMuLV LTR was unsatisfactory after transduction of the PLB985 X-CGD knockout cell line and of primary CD34+ hematopoietic progenitors from X-CGD patients. The presence of either constitutive or inducible internal promoters did not result in important improvements in the efficiency of O2- production and lowered the titers of the viral preparations. In contrast, sustained levels of superoxide generation were obtained upon transduction with the FMEV vector. To analyze the efficiency of transgene expression at the single cell level, over 150 cellular clones were generated from bulk cultures of PLB985 X-CGD cells transduced with this vector, each one representative of an individual transduction event. These clones revealed a markedly heterogeneous pattern of gp91-phox expression, ranging from complete silencing to full restoration of superoxide production. Within each clone, expression of the therapeutic gene correlated with the number of expressing cells rather than with the average levels of expression from each cell, indicating that at the single cell level, the proviral promoter is regulated by a binary, on/off mechanism. Moreover, both transduced bulk and clonal cell populations displayed a tendency to a progressive extinction of expression over time, with a mechanism involving LTR methylation. The design of novel retroviral vectors escaping silencing is highly desirable for efficient gene therapy. PMID- 10673721 TI - Type I consensus interferon (CIFN) gene transfer into human melanoma cells up regulates p53 and enhances cisplatin-induced apoptosis: implications for new therapeutic strategies with IFN-alpha. AB - In this study, we describe the effects produced by the retroviral transduction of human type I consensus IFN (CIFN) coding sequence into the 8863 and 1B6 human melanoma cell lines, derived from a metastatic and a primary human melanoma, respectively. Melanoma cell lines producing approximately 103 IU/ml of IFN were obtained. Interestingly, cisplatin treatment of IFN-producing 8863 and 1B6 melanoma cells resulted in a three- to four-fold increase in the percentage of apoptotic cells with respect to similarly treated parental or control-transduced cell cultures. A similar effect, although less intense, was caused by cultivation of parental melanoma cells in the presence of exogenous CIFN. The increased susceptibility of the IFN-producing melanoma cell lines to cisplatin-induced apoptosis was associated with an IFN-dependent accumulation of p53, which also correlated with a decrease in Bcl-2 expression. Addition of exogenous CIFN to parental melanoma cells resulted in similar although weaker modulations of p53 and Bcl-2 expression. Cisplatin administration to nude mice bearing 3-day-old IFN producing 8863 tumors resulted in complete tumor regression, while only a partial tumor inhibition was observed upon cisplatin treatment of mice bearing parental or control-transduced 8863 tumors. Starting the cisplatin treatment 7 days after tumor cell injection still resulted in a stronger inhibition of tumor growth in the mice bearing IFN-producing 8863 tumors as compared with parental tumor bearing mice. A comparable therapeutic effect was obtained after repeated peritumoral administration of 103 IU of exogenous CIFN and cisplatin treatment. Interestingly, a spontaneous tumor regression was observed in nude mice injected with IFN-producing 1B6 cells, in contrast to the progressive tumor growth occurring in mice receiving a similar inoculum of the parental or control transduced 1B6 melanoma cells. Repeated peritumoral administration of 103 IU of exogenous CIFN to mice bearing parental 1B6 tumors caused only a transient inhibition of tumor growth. These results indicate that type I IFN gene transfer is an effective approach for suppressing the tumorigenic phenotype of human melanoma cells and for increasing the efficacy of anticancer drugs. These observations, together with our previous findings showing the importance of IFN alpha-T cell interactions in the generation of an antitumor response in mouse models, underline the interest of using type I IFN in gene therapy strategies for the treatment of human melanoma. PMID- 10673723 TI - Publisher's announcement PMID- 10673722 TI - The Journal of Human Hypertension and the millennium. PMID- 10673724 TI - Myocardial texture in hypertrophic remodelling: new insight into ventricular load and function? PMID- 10673725 TI - Ultrasonic myocardial textural parameters and midwall left ventricular mechanics in essential arterial hypertension. AB - BACKGROUND: The evaluation of the systolic left ventricular performance in hypertensive patients presents some problems related to left ventricular hypertrophy (LVH) which alters the ventricular geometry. The videodensitometric textural ultrasonic analysis of hypertensive myocardium has provided evidence of impairment in the cyclic variation of the mean gray level. This might be considered as an index of intrinsic myocardial function. OBJECTIVES: The aim of the present study was to analyse the connection between the midwall fractional shortening and end-systolic stress. The ultrasonic textural parameters in hypertensive patients, arranged in different groups according to the level of LVH and relative wall thickness, were also evaluated. METHODS: A group of age-matched (58 +/- 7 years) male essential hypertensive patients (n = 70) were compared to a group of normotensive and healthy subjects used as controls (n = 32). All subjects performed a conventional 2D-Doppler echocardiography to analyse the left ventricular performance. A quantitative analysis of the echocardiographic digitised imaging was also carried out with the help of a calibrated digitization system in order to calculate the septum and the posterior wall textural parameters. The myocardial mean gray level was calculated to derive the cyclic variation index (CVI). RESULTS: When subjected to a higher meridional end systolic stress, the hypertensive patients showed a significantly lower midwall fractional shortening than the control patients. The CVI was also significantly lower in the hypertensives group, both for the septum wall (-16.3 +/- 22.8 vs34.7 +/- 15.3%; P < 0.001) and the posterior wall (-5.2 +/- 23.6 vs 38.2 +/- 15.4%; P < 0.001). A significant correlation was found between the midwall fractional shortening (MFS) and the textural parameters, and between these two variables and the end-systolic stress. CONCLUSION: The CVI was found to be a highly sensitive parameter in the identification of abnormal echodensity in essential hypertension. The CVI was significantly lower in patients with concentric hypertrophy in comparison with other left ventricular geometric models. This parameter could be considered as an index of the intrinsic myocardial function, being related, in essential hypertension, to midwall fractional shortening and to end-systolic stress. Journal of Human Hypertension (2000) 14, 9-16. PMID- 10673726 TI - Beta-adrenergic receptor density and function in left ventricular hypertrophy in young essential hypertensives. AB - A sympathetic overactivity has been reported in the early stages of essential hypertension and has been involved in the pathogenesis of left ventricular hypertrophy (LVH) in essential hypertension. The state of beta2-adrenergic receptors as related to the presence of this complication was investigated in a group of 15 essential hypertensive patients and compared to 10 normotensive control subjects. Left ventricular mass index was determined by bidimensional echocardiography. Plasma catecholamine levels were measured by a radioenzymatic assay. beta2-adrenoceptor density was measured in intact lymphocytes by radioligand binding assay, using the hydrophilic ligand CGP 12177. beta2- adrenoceptor function was assessed by measuring intracellular cAMP levels in isoproterenol-stimulated lymphocytes. Left ventricular mass index (P < 0.05), body mass index (P < 0.01), plasma noradrenaline levels (P < 0.05) and beta2 adrenoceptor density (P < 0.05) were higher in hypertensives than in controls. Left ventricular mass index correlated with body mass index both in normotensives and hypertensives, as well as with plasma noradrenaline levels only in normotensives. Left ventricular mass index also showed a positive correlation with mean arterial pressure and an inverse relationship with beta2-adrenoceptor density and response only in hypertensive patients. In conclusion, left ventricular hypertrophy in young essential hypertensives is associated to a reduced beta2-adrenoceptor density and function, probably as a compensating mechanism of the hypertrophied myocardiocyte secondary to the increased sympathetic outflow. Journal of Human Hypertension (2000) 14, 17-21. PMID- 10673727 TI - Relationships between cardiovascular remodelling and the pulse pressure in never treated hypertension. AB - The role of pulse pressure (PP) in cardiovascular remodelling was studied in 61 never treated hypertensive subjects who were selected on the criteria of ambulatory blood pressure (BP) monitoring (mean BP over 24 h: 147 +/- 14/96 +/- 10 mm Hg). Echocardiography and carotid ultrasonography were performed and the vascular images analysed using a specific automatic measuring program. Thirty percent of subjects had left ventricular hypertrophy (LVH). Left ventricular mass index (LVMI) was related to the clinic (r = 0.35) and ambulatory (r = 0.41 over 24 h, r = 0.38 daytime and r = 0.42 night-time) PP and to the systolic BP. PP was higher when there was LVH. Vascular thickening was found in 6.6% of subjects (carotid intima-media thickness (IMT) >/=1.0 mm). Among the BP parameters, IMT and cross-sectional area (CSA) were related only to the clinic PP (r = 0.27, r = 0.29 respectively) and to the ambulatory PP (over 24 h: r= 0.29, r = 0.28; daytime: r = 0.22, r = 0.23; night-time: r = 0.32, r = 0.30). In men, the relationship between CSA and PP (clinic and over 24 h) was independent of age. A total of 16.7% of subjects with LVH had intima-media thickening in contrast to 2.3% in the group without LVH. LVMI was related to the CSA (r = 0.37) and to the IMT (r = 0.31). However, after multivariate analysis taking into account the PP, relationships between IMT or CSA and LVMI disappeared. Our data showed that the PP was the most important BP parameter in the development of cardiac and arterial remodelling in hypertension. Journal of Human Hypertension (2000) 14, 23-30. PMID- 10673728 TI - The mercury sphygmomanometer should be abandoned before it is proscribed. AB - Both in clinical practice and medical research, blood pressure is still largely measured by auscultation using a mercury sphygmomanometer. Blood pressure is the most important predictor of life expectancy. Treatment of high blood pressure reduces strokes, heart attack and heart failure. Accurate measurement is therefore essential. At a large London teaching hospital, just under 500 mercury sphygmomanometers and their associated cuffs were examined. More than half had serious problems that would have rendered them inaccurate in measuring blood pressure. At the same time, assessment of the technical knowledge needed to measure blood pressure by the ausculatory technique was also carried out amongst medical and nursing staff. This showed a considerable level of ignorance. These results inevitably lead to inaccurate measurement of blood pressure with serious consequences. In addition mercury is a non-degradable pollutant, eventually accumulating on the sea bed. The use of mercury in sphygmomanometers is already in the process of being eliminated in Scandinavia and Holland and other countries are likely to follow. Our results suggest that mercury sphygmomanometers are not adequately maintained and require expertise that is not available for accurate measurement of blood pressure. Their use should be dispensed with on these grounds before a ban for other and, perhaps less justifiable reasons. Validated automatic devices, which are less liable to measurement and observer error should be used instead. At the same time a concerted effort is needed to instruct health care professionals on the importance of more accurate measurement of blood pressure. Journal of Human Hypertension (2000) 14, 31-36. PMID- 10673729 TI - In-field validation of automatic blood pressure measuring devices. AB - The method for rapid evaluation of automatic blood pressure (BP) measurement devices (READ) is based on numerous BP measurements at rest and during a standardised postural challenge in a small number of subjects with a wide range of BPs. The present study proposes additional parameters of the READ for in-field validation of automatic BP measurement devices. BP measurements were done in supine position for 10 min followed by head-up tilt for 30 min and again supine for 10 min. BPs were determined simultaneously by automatic (AU) and mercury sphygmomanometric (MS) measurements on the same arm. The BP differences DeltaBP:AU-MS were calculated. Three units of the Colin BP-8800 and the Datex Engstrom Cardiocaptrade mark II were evaluated. Based on DeltaBP(AU-MS), the grade of accuracy, aberration patterns and correlates of accuracy were assessed for each unit. Per unit, an average of 121 measurements were done, every BP category being met in >/=20 MS measurements. In general, the AU systolic BP values were higher than MS systolic BP values (mean systolic DeltaBP = 1.26 +/- 17.1 mm Hg) and AU diastolic BP values were lower than MS diastolic BP values (mean diastolic DeltaBP = -12.31 +/- 7.8 mm Hg). All units classified into category D of the British Hypertension Society grading system and exhibited inconsistent aberration patterns, making design of correction formulas impractical. Inaccuracy of the instruments was independent on mode of measurement, posture, magnitude of the BP and heart rate, early or late measurements from beginning of the head-up tilt test, and prolonged use of the unit. The READ permitted to identify rapidly the degree of accuracy of automatic BP measuring devices. Identification of the aberration pattern of an instrument could be the basis for calculating equations for correction of the measured BP. Further studies will show which parameters of the READ may expose specific defects of the instruments. Journal of Human Hypertension (2000) 14, 37-42. PMID- 10673730 TI - Role of the Gly460Trp polymorphism of the alpha-adducin gene in primary hypertension in Scandinavians. AB - Previous studies have suggested that the Trp460 allele of the Gly460Trp polymorphism in the alpha-adducin gene is associated with salt sensitivity and primary hypertension. The present study was undertaken to evaluate if the Trp460 allele of this polymorphism is associated with primary hypertension in Scandinavians. To address this issue, 294 patients with primary hypertension and 265 normotensive control subjects from Sweden were examined and genotyped for the Gly460Trp polymorphism using polymerase chain reaction and restriction fragment length polymorphism methods. We then used a population of 80 patients with primary hypertension and 154 normotensive control subjects from Finland to replicate the findings. The frequency of the Trp460 allele was lower in hypertensive patients than in normotensive controls in the Swedish population (17.7% vs 23.0%; P = 0.03) and in the Finnish population (14.4% vs 19.5%; NS). Therefore we also performed a pooled analysis in which the frequency of the Trp460 allele was significantly lower in hypertensive patients than in normotensive controls (17.0% vs 21. 7%; P = 0.02). In subjects who did not receive antihypertensive medication (n = 447) there was no difference between carriers of the three different codon 460 genotypes (Trp-Trp; Trp-Gly and Gly Gly) either for systolic (128 +/- 18; 127 +/- 15 and 129 +/- 17 mm Hg, NS) or for diastolic blood pressure (75.6 +/- 12.1; 74.7 +/- 9.3 and 75.0 +/- 10.4 mm Hg, NS). In conclusion, the lower frequency of the Trp460 allele in hypertensive patients than in normotensive controls strongly argues against a pathogenic role of this allele in primary hypertension. The results rather suggest that another variant in linkage disequilibrium with the Gly460Trp polymorphism increases susceptibility for hypertension. Journal of Human Hypertension (2000) 14, 43-46. PMID- 10673731 TI - Lack of association between ACE gene polymorphism and left ventricular hypertrophy in essential hypertension. AB - The possible association between the insertion/deletion (I/D) polymorphism of the angiotensin I converting enzyme (ACE) gene and left ventricular hypertrophy (LVH) was investigated in a group of essential hypertensive patients. Seventy-one essential hypertensive patients (35 men and 36 women), aged 51 +/- 1 years, were genotyped by PCR for the I/D polymorphism of the ACE gene. Cardiac morphology and function were assessed by means of M-mode echocardiography. The relative frequencies of the three genotypes, DD, DI, and II, were respectively: 24%, 55%, and 21%. Mean values of left ventricular mass index were 145, 144, and 150 g/m2 for DD, DI, and II genotypes, without significant differences among them (P = 0.82). Likewise, the prevalence of LVH (76%, 64%, and 87%) was not significantly different among the three genotypes (P = 0.23). We conclude that the ACE gene I/D polymorphism is not associated with LVH in essential hypertension. Journal of Human Hypertension (2000) 14, 47-49. PMID- 10673732 TI - Differences in insulin sensitivity and risk markers due to gender and age in hypertensives. AB - Men run a higher risk for cardiovascular disease than women, even if hypertensive. This has been attributed to a more pronounced central (abdominal) fat distribution in men as well as menopausal state in women. The hypothesis to be tested in hypertensives was that men have more pronounced insulin resistance and other cardiovascular risk factors than pre-menopausal, but not post menopausal, women. We carried out a cross-sectional observation study of middle aged hypertensives of both sexes, divided into two age groups, below or over 50 years of age. The study was performed in untreated out-patients, visiting a hypertension policlinic, in Uppsala, Sweden. Three hundred men and 170 women with a mean age of 57 years were investigated. Measurements were taken by: physical examination (body mass index, waist-to-hip ratio, blood pressure); intravenous glucose tolerance test (IVGTT); euglycaemic hyperinsulinaemic clamp; and blood sampling for lipoprotein lipid fractions, uric acid, and free fatty acids. The results were that pre-menopausal women showed a higher insulin-mediated glucose disposal (7.6 vs5.8 mg/kg/min; P < 0. 01), and lower fasting glucose (4.9 vs 5.2 mmol/l; P < 0.05) than men, as well as a more advantageous lipoprotein profile. However, in post menopausal women insulin sensitivity decreased and the lipoprotein profile deteriorated. Women still showed higher levels of high density lipoprotein (HDL)-cholesterol, and men a higher waist-to-hip ratio and levels of uric acid, in both age groups. It was concluded that post-menopausal hypertensive women are relatively more insulin resistant than pre-menopausal ones in comparison with men in the same age group and with the same degree of overall obesity. Journal of Human Hypertension (2000) 14, 51-56. PMID- 10673733 TI - Salt-induced exacerbation of morning surge in blood pressure in patients with essential hypertension. AB - The morning surge in blood pressure (BP) is related to the morning occurrence of lethal cardiovascular events. We tested the hypothesis that salt intake may be associated with the morning surge in BP in essential hypertension. Seventy-six patients were admitted and placed on a low salt diet (2 g/day) for 7 days followed by a high salt diet (20-23 g/day) for another 7 days. At the end of each salt diet, 24-h ambulatory BP and heart rate monitorings and head-up tilt (HUT) test were performed. Patients whose average mean BP (MBP) was increased by more than 10% by salt loading were assigned to the salt-sensitive (SS) group (n = 37); the remaining patients, whose MBP was increased by less than 10%, were assigned to the non-salt-sensitive (NSS) group (n = 39). The increase in ambulatory MBP during 6.30-8.00 am above the baseline (2.00-4.00 am) was significantly enhanced by salt loading in the NSS group (P < 0.05), but not in the SS group. The coefficient of variation of 24-h MBP and heart rate was increased by salt loading only in the NSS group. The significant elevation of plasma noradrenaline concentration after awakening, which was noted during the low salt diet period, was unchanged during the high salt diet period in the NSS group, but abolished in the SS group. Salt loading enhanced HUT-induced decrease in systolic BP without affecting the heart rate response only in the NSS group. We conclude that the morning surge in BP is enhanced by salt loading in the NSS type of essential hyper- tension, presumably by the excessive activation of the sympathetic nervous system. Journal of Human Hypertension (2000) 14, 57-64. PMID- 10673734 TI - Correlates of blood pressure in an urban Zimbabwean population and comparison to other populations of African origin. AB - We have evaluated the relationship between systolic blood pressure (SBP) and age, body mass index (BMI), waist circumference, sodium to potassium ratio (Na/K), and tobacco use in an urban African population. We conducted a random, population based, cross-sectional survey of people 25 years and older in Marondera, Zimbabwe, with over-sampling in older age groups (n = 775), using a method comparable to that used in International Collaborative Study on Hypertension in Blacks (ICSHIB). The age-adjusted prevalences of hypertension in Marondera (SBP >/=140/DBP >/=90/antihypertensive medication) were 30% for women and 21% for men. The average BMI was 26.3 kg/m2 for women and 21.4 kg/m2 for men. The prevalence of hypertension had a steep association with age and in women ranged from 15% (25 34 years) to 63% (55 years and over) and in men from 9% to 47%. No tobacco use in women and greater Na/K ratio in spot urines in men were significantly associated with an increased SBP. In both men and women the levels of hypertension and SBP were strongly positively associated with BMI, although the relationship appeared to plateau in women with a BMI greater than >/=25 kg/m2. At a given BMI, men and women had similar SBPs and prevalences of hypertension. There is a very high prevalence of hypertension among urban Zimbabweans, particularly among women. Under the assumption the studies are comparable, the prevalence of hypertension in Zimbabwean women (41%) and men (26%) after age adjustment to the ICSHIB populations, appeared higher than almost all of the ICSHIB populations, including those with higher average body mass indexes. Journal of Human Hypertension (2000) 14, 65-73. PMID- 10673735 TI - Does the Dundee Step Test predict outcome in treated hypertension? A sub-study protocol for the ASCOT trial. Anglo-Scandinavian Cardiac Outcome Trial. AB - Treated hypertensive subjects may remain five times more likely to die of cardiac and cerebrovascular diseases than normotensive subjects with equivalent resting blood pressure (BP) levels. Research evidence suggests that exercise BP is a better predictor of end-organ damage and mortality than resting BP, and data from our centre show that a significant proportion of treated hypertensives have uncontrolled BP during a 5-min Dundee Step Test. The prognostic usefulness of exercise BP has yet to be translated into clinical practice because of the lack of a suitable technique. The Dundee Step Test is being evaluated in the ASCOT (Anglo-Scandinavian Cardiac Outcome Trial) study, a 5-year follow-up multicentre, multinational trial comparing the effect of newer (amlodipine and perindopril) and older (bendroflumethiazide and atenolol) antihypertensive agents stratified according to cholesterol levels on cardiac outcome. If the value of the Dundee Step Test is proven, then it may be adopted into routine clinical practice for the assessment of exercise BP. This may result in the improved management of hypertension with a subsequent reduction in morbidity and mortality. The publication of this study protocol is meant to be a statement of on-going research which may stimulate interest among those with an interest in this area of research. Journal of Human Hypertension (2000) 14, 75-78. PMID- 10673736 TI - Individualized methotrexate dosing in children with relapsed acute lymphoblastic leukemia. AB - Although high-dose methotrexate has been extensively studied in children with newly diagnosed acute lymphoblastic leukemia (ALL), there are fewer data in children with relapsed ALL, many of whom have been heavily pretreated and have subclinical kidney dysfunction. We characterized the pharmacokinetics of adaptively controlled methotrexate given as a 24-h infusion during consolidation therapy in 24 children with relapsed ALL. To achieve the target steady-state concentration of 65 microM, dosage adjustments were required in 14 patients, with doses ranging from 2854 to 6700 mg/m2 per course. The mean steady-state plasma concentration (Cpss) of 68.0 microM was different (P = 0.025) than the predicted Cpss (mean = 87.4 microM; range 35.7-184 microM) had no adjustment in dose been made. The coefficient of variation in Cpss was reduced from 41% to 18% by individualizing doses. Predisposing factors that correlated with decreased methotrexate clearance were female sex (P = 0.03), age greater than 6 years (P = 0.01), and prior history of heavy amphotericin B treatment (>30 mg/kg) (P = 0.03), but no factor predicted low clearance as well as the measured initial methotrexate clearance during the infusion (P < 0.0001). There was no life threatening toxicity with the regimen. We conclude that dosage individualization decreases interpatient variability and avoids potentially toxic methotrexate exposures in heavily pretreated ALL patients. PMID- 10673737 TI - Multivariate analysis of prognostic factors in patients with refractory and relapsed acute myeloid leukemia undergoing sequential high-dose cytosine arabinoside and mitoxantrone (S-HAM) salvage therapy: relevance of cytogenetic abnormalities. AB - To improve the basis for the stratification of patients with refractory and relapsed acute myeloid leukemia (AML) univariate and multivariate analyses of prognostic factors were performed in 254 patients (median age 50 years, range 18 74) undergoing S-HAM salvage chemotherapy during two consecutive prospective trials of the German AML Cooperative Group. In a multivariate analysis, duration of the first complete remission (CR) was the only factor associated with time to treatment failure (P = 0.0223). Disease-free survival was influenced by a short duration of the first CR of less than 6 months (P = 0.0001), WBC (P = 0.0018), blast count (P = 0.0037), and neutrophil count (P = 0.0119). The achievement of CR was related to the hemoglobin level only (P = 0.0457), the early death rate was related to age only (P = 0.0109), and survival was related to the bilirubin level only (P = 0.0166). In the subgroup of 104 patients in whom additional karyotype analyses were performed prior to first-line therapy unfavorable chromosome abnormalities were associated with a lower CR rate (univariate analysis, P = 0.0342; CR 24% vs 53%) and were the only factor related to survival. These analyses warrant the further evaluation of the impact of cytogenetic abnormalities on the outcome of patients with advanced AML in order to improve the characterization according to duration of first CR and to WBC of distinct subgroups of patients with differing prognoses as a basis for stratification in future trials. PMID- 10673738 TI - Glutathione S-transferase genotypes in children who develop treatment-related acute myeloid malignancies. AB - Epipodophyllotoxin-associated secondary myeloid leukemia is a devastating complication of acute lymphoblastic leukemia (ALL) therapy. The risk factors for treatment-related myeloid leukemia remain incompletely defined. Genetic deficiencies in glutathione S-transferase (GST) activities have been linked to higher frequencies of a number of human malignancies. Our objective was to determine whether the null genotype for GSTM1, GSTT1, or both, was more frequent in children with ALL who developed treatment-related myeloid malignancies as compared to those who did not. A PCR technique was used to assay for the null genotype for GSTM1 and GSTT1 in 302 children with ALL, 57 of whom also subsequently developed treatment-related acute myeloid leukemia or myelodysplastic syndrome. Among children with ALL who did not develop treatment related myeloid malignancies, the frequencies of GSTM1 and GSTT1 wild-type, GSTM1 null-GSTT1 wild-type, GSTM1 wild-type-GSTT1 null, and GSTM1 and GSTT1 null genotypes were 40%, 42%, 9% and 9%, respectively. The corresponding frequencies for patients who developed acute myeloid malignancies were 42%, 32%, 11% and 16%, respectively (P = 0.26). A statistically significant increase in the frequency of the GST null genotype was observed in male patients who developed myeloid malignancies as compared to male ALL control patients (P = 0.036), but was not observed in female patients (P = 0.51). Moreover, a logistic regression analysis of possible predictors for myeloid malignancies, controlling for gender and race, did not reveal an association of GSTM1 or GSTT1 null genotypes (P = 0.62 and 0.11, respectively) with treatment-related malignancies. Our data suggest that GSTM1 and GSTT1 null genotypes may not predispose to epipodophyllotoxin associated myeloid malignancies. PMID- 10673739 TI - Acute lymphoblastic leukemia with an unusual t(8;14)(q11.2;q32): a Pediatric Oncology Group Study. AB - We present the clinicopathologic findings and survival data on 10 patients with acute lymphoblastic leukemia (ALL) and a rare t(8;14)(q11.2;q32). There were five male and five female patients, nine Caucasians and one Black, aged 4-17 (median 10.9) years. Three had Down syndrome. Eight (80%) patients had a white blood cell (WBC) count <50 x 109/l at presentation. No patient had central nervous system involvement or a mediastinal mass. Two patients had concurrent splenomegaly and hepatomegaly. Adenopathy was absent in four, minimal in three, moderate in one and prominent in two patients. All eight cases where immunophenotyping was performed by flow cytometry showed a B-precursor phenotype with expression of CD10 (CALLA). Only one case exhibited t(8;14)(q11.2;q32) as the sole karyotypic abnormality. Three patients were classified as standard-risk and seven high-risk by NCI (National Cancer Institute) consensus risk group categories. All patients achieved complete remission and seven patients were in complete continuous remission (CCR) after chemotherapy designed for B-precursor ALL. Three patients relapsed after 23.5, 31.3 and 32.1 months of EFS; the first patient also had t(9;22)(q34;q11), the second had a WBC count of 126 x 109/l at presentation while the third patient had no high risk features except for age 10 years. Thus, from our data, the t(8;14)(q11.2;q32) does not appear to confer an increased risk of relapse. Further observations are needed to confirm this conclusion. PMID- 10673740 TI - Monosomy 20 as a pointer to dicentric (9;20) in acute lymphoblastic leukemia. AB - Twenty new cases of acute lymphoblastic leukemia (ALL) with the dicentric chromosome dic(9;20)(p1113;q11) are presented. This chromosomal abnormality is difficult to identify from G-banding alone. It masquerades as monosomy 20 and is only accurately identified by fluorescence in situ hybridization (FISH). Monosomy 20 was found in 59/2790 patients with successful karyotypes entered to the Leukaemia Research Fund/UK Cancer Cytogenetics Group Karyotype Database in ALL (LRF/UKCCG Karyotype Database). FISH revealed dic(9;20) in 20/25 cases with available material. Extra copies of chromosome 21 were found in 8 of the 20 cases. Patients were 14 females and six males, aged 1-32 years (median 4 years), with leukocyte counts of 2-536 (median 23) x 109/l and immunophenotypes of common or pre-B ALL (17 cases), T-ALL (one case) or unknown (two cases). Four patients relapsed at 2, 22, 28 and 47 months and two died at 49 and 63 months (median follow-up 37 months). FISH studies on the remaining five patients showed one with monosomy 20 and four with other rearrangements of the chromosome. This study has increased the number of reported cases of dic(9;20) from 17 to 37. It has identified dic(9;20) in one case of T-ALL and shows an association of this translocation with trisomy 21. PMID- 10673741 TI - Minimal residual disease is common after allogeneic stem cell transplantation in patients with B cell chronic lymphocytic leukemia and may be controlled by graft versus-host disease. AB - Following allogeneic stem cell transplantation (SCT), we studied the presence of donor and recipient derived cells within the CD19+ B cell fraction, in patients with B cell chronic lymphocytic leukemia (CLL). The chimeric status of the six patients studied was further investigated with minimal residual disease (MRD) detection, by sequencing and using patient-specific primers derived from junctional regions of clonally rearranged immunoglobulin heavy-chain (IgH) receptor genes. To date, five of six patients are alive with a median follow-up time of 24 months (range 15-60) post-SCT. All patients experienced acute and chronic graft-versus-host disease and responded clinically to SCT. All patients were MRD positive after SCT, which correlated to mixed chimerism within the CD19+ cell fraction in all samples except one (25/26). High levels of tumor necrosis factor-alpha (TNF-alpha) and soluble interleukin-2 receptor (sIL-2R) indicated advanced disease, and patients with increased levels pre- and post-SCT were also those with the most long-lasting PCR-detectable MRD post-SCT. Hence, a high tumor burden pre-SCT may reflect the long duration of detectable MRD in patients with B CLL after SCT. A durable anti-leukemic effect was probably important in these patients. PMID- 10673742 TI - A mutated PML/RARA found in the retinoid maturation resistant NB4 subclone, NB4 R2, blocks RARA and wild-type PML/RARA transcriptional activities. AB - The fusion protein PML/RARA, associated with acute promyelocytic leukemia behaves as an abnormal retinoic acid (RA) receptor with altered transactivation properties but is still inducible by RA. The chimeric protein is thought to promote leukemogenesis but also paradoxically to mediate the sensitivity to ATRA of APL cells. This has been supported by works reporting that in vitro ATRA resistance is characterized by defects in the RARA/E-domain of PML/RARA. In the present report, we identified a new mutation in the E domain of PML/RARA which is associated with a RA-resistant subline of NB4 cells; NB4-R2. This mutation, identical to the Gln411 mutation found in HL60-R, changes the amino acid Gln903 to an in-phase stop codon, generating a truncated form of PML/RARA which has lost 52 amino acids at its C-terminal end. We have studied the effect of the truncated PML/RARA protein on PML NB formation and RARA and PML/RARA transcriptional activity. We show here that the fusion mutant exerts a dominant negative effect on wild-type PML, PML/RARA and RARA transcription activity. These findings highlight the important role of the RARA E-domain of PML/RARA in mediating RA sensitivity in APL cells. PMID- 10673743 TI - Arsenic trioxide-induced apoptosis and differentiation are associated respectively with mitochondrial transmembrane potential collapse and retinoic acid signaling pathways in acute promyelocytic leukemia. AB - Recent studies showed that arsenic trioxide (As2O3) could induce apoptosis and partial differentiation of leukemic promyelocytes. Here, we addressed the possible mechanisms underlying these two different effects. 1.0 microM As2O3 induced apoptosis was associated with condensation of the mitochondrial matrix, disruption of mitochondrial transmembrane potentials (DeltaPsim) and activation of caspase-3 in acute promyelocytic leukemia (APL) cells regardless of their sensitivity to all-trans retinoic acid (ATRA). All these effects were inhibited by dithiothreitol (DTT) and enhanced by buthionine sulfoximine (BSO). Furthermore, BSO could also render HL60 and U937 cells, which had the higher cellular catalase activity, sensitive to As2O3-induced apoptosis. Surprisingly, 1.0 microM As2O3 did not induce the DeltaPsim collapse and apoptosis, while 0.1 microM As2O3 induced partial differentiation of fresh BM cells from a de novo APL patient. In this study, we also showed that 0.2 mM DTT did not block low-dose As2O3-induced NB4 cell differentiation, and 0. 10.5 microM As2O3 did not induce differentiation of ATRA-resistant NB4-derived sublines, which were confirmed by cytomorphology, expression of CD11b, CD33 and CD14 as well as NBT reduction. Another interesting finding was that 0.10.5 microM As2O3 could also induce differentiation-related changes in ATRA-sensitive HL60 cells. However, the differentiation-inducing effect could not be seen in ATRA-resistant HL60 sublines with RARalpha mutation. Moreover, low-dose As2O3 and ATRA yielded similar gene expression profiles in APL cells. These results encouraged us to hypothesize that As2O3 induces APL cell differentiation through direct or indirect activation of retinoic acid receptor-related signaling pathway(s), while DeltaPsim collapse is the common mechanism of As2O3-induced apoptosis. PMID- 10673744 TI - Molecular profile of Epstein-Barr virus infection in HHV-8-positive primary effusion lymphoma. AB - Primary effusion lymphoma (PEL) selectively involves the serous body cavities, occurs predominantly in immunodeficient patients and is infected consistently by human herpesvirus type-8. PEL is also frequently infected by Epstein-Barr virus (EBV). The precise pathogenetic role of EBV coinfection in PEL is not fully understood. The lymphoma fails to express the EBV transforming proteins EBNA-2 and LMP-1, whereas it expresses EBNA-1 (latency I phenotype). Some studies have hypothesized that other EBV-positive lymphomas expressing the latency I phenotype may associate with specific molecular variants of EBNA-1, although this issue has not been addressed in PEL. On this basis, this study is aimed at a detailed molecular characterization of EBV in PEL. Fifteen EBV positive PEL (12 AIDS related, one post-transplant, two arising in immunocompetent hosts) were subjected to molecular characterization of the viral genes EBNA-1 and LMP-1, as well as definition of EBV type-1/type-2. The EBNA-1 gene displayed a high degree of heterogeneity in different cases of PEL, with seven distinct recognizable variants and subvariants. A wild-type LMP-1 gene was detected in 10/15 cases, whereas in 5/15 cases the LMP-1 gene harbored a deletion spanning codons 346-355. EBV type-1 occurred in 11/15 PEL whereas EBV type-2 occurred in 4/15 cases. Despite a high degree of genetic variability of the virus in different PEL cases, each single PEL harbored only one EBV variant, consistent with monoclonality of infection and suggesting that infection preceded clonal expansion. Overall, our results indicate that: (1) individual PEL cases consistently harbor a single EBV strain; (2) EBNA-1 displays a high degree of heterogeneity in different PEL cases; (3) no specific EBV genotype preferentially associates with PEL. PMID- 10673745 TI - Spontaneous reduction of leukemic lymphoma cells possibly by anti-tumor antibody mediated phagocytosis; a kappa lambda-dual-positive B cell lymphoma. AB - We have investigated the possible role of anti-tumor antibody detected in a case of follicular lymphoma which demonstrated the spontaneous reduction of leukemic tumor cells. The tumor cells genotypically had monoclonal rearrangements of the immunoglobulin J H and C kappa genes, but phenotypically exhibited surface IgG, A, kappa and lambda (kappa lambda dual positivity). The culture study revealed that IgGlambda, at least, was derived from the serum, and IgAkappa was expressed intrinsically. Furthermore, the positive correlation between the densities of both surface light chains on two-color flow cytometry, the rosette formation study and its inhibition test by the Fcgamma fragment suggested that the serum IgGlambda combined with some antigens on the tumor-cell surface via its Fab portion and with the Fcgamma receptor of macrophages via its Fc portion. From these findings, we regarded the present case as an anti-tumor antibody-coated lymphoma. In addition, the phagocytic study disclosed that the serum-derived IgGlambda, at least, might have induced the phagocytosis of circulating lymphoma cells by macrophages. In conclusion, the existence of the anti-tumor antibody coated lymphoma may be helpful in clarifying the immunological mechanism of the spontaneous regression occasionally seen in lymphomas. PMID- 10673746 TI - The tyrosine kinase receptor met and its ligand HGF are co-expressed and functionally active in HHV-8 positive primary effusion lymphoma. AB - Primary effusion lymphoma (PEL) harbors consistent infection by human herpesvirus 8, preferentially develops in immunodeficient patients and selectively localizes to the serous body cavities. Histogenetic analysis has suggested that PEL originates from post-germinal center, pre-terminally differentiated B cells sharing phenotypic features with plasma cells. Here we have investigated the expression status and functional integrity of the Met tyrosine kinase receptor and of its ligand hepatocyte growth factor (HGF). Thirteen PEL (nine cell lines and four primary specimens) were analyzed for Met and HGF expression and function by multiple assays. For comparison, a panel of 34 high grade B cell non-Hodgkin lymphomas (NHL) other than PEL was also investigated. Co-expression of Met and HGF was found in all PEL analyzed, whereas it was restricted to 1/34 B cell NHL other than PEL (P < 0.001; chi2 test). The Met protein expressed by PEL displays biochemical characteristics typical of Met expressed by other cell types and is capable of tyrosine autophosphorylation. By using a combination of immunological and biological assays, production and secretion of a functional HGF species was identified in all PEL cell lines analyzed. HGF stimulation of PEL cells rapidly induces Met tyrosine phosphorylation, demonstrating the functional integrity of the Met/HGF loop. Because of the well known mitogenic and motogenic properties of Met/HGF interactions, these data may bear implications for PEL growth and dissemination. Among B cell neoplasms, Met/HGF co-expression selectively clusters with PEL and, as demonstrated by previous studies, with multiple myeloma plasma cells, thus reinforcing the notion that PEL displays biologic similarities with tumors derived from late stages of B cell differentiation. PMID- 10673747 TI - Proliferative response of mantle cell lymphoma cells stimulated by CD40 ligation and IL-4. AB - Mantle cell lymphoma (MCL) is a tumor of intermediate-size, IgM+, IgD+ B cells derived from the mantle zone of the germinal center. Little is known about its specific immunologic features or responsiveness to T cell-derived signals. In this work, we evaluated the proliferation and cell cycle properties of freshly isolated MCL cells after CD40 ligation, in the absence and presence of interleukin 4 (IL-4). In each MCL case examined, there was a marked growth enhancing effect of these two stimuli characterized by improved viability, augmented expression of Ki-67, and induction of the proliferating cell nuclear antigen (PCNA). Cyclin D1 was expressed throughout the cell cycle in MCL cells induced to enter S phase. From these investigations, we conclude that the biology of MCL B lymphocytes is affected by CD154 (CD40 ligand) and IL-4, two signals usually provided by CD4+ T cells. The capacity to manipulate the activation and cell cycle state of MCL cells by these specific immunological stimuli may be exploited to confer susceptibility to chemotherapy agents and develop novel therapies in this disease. PMID- 10673748 TI - 2-Chloro-2'-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4. AB - The mechanism of apoptosis induced by 2-chloro-2'-deoxyadenosine (2CdA) in human leukemia cell line MOLT-4 was investigated. 2CdA induced increases of 3'-OH ends of genomic DNA, ladder-like DNA fragmentation and phosphatidylserine translocation to the outer membrane, which are apoptotic characteristics. These apoptotic phenomena induced by 2CdA were inhibited by cycloheximide (CHX; a protein synthesis inhibitor), deoxycytidine (dC; a substrate of deoxycytidine kinase), acetyl Ile-Glu-Thr-Asp aldehyde (Ac-IETD-CHO; a caspase-8 inhibitor) and acetyl Asp-Glu-Val-Asp aldehyde (Ac-DEVD-CHO; a caspase-3 inhibitor). The protein synthesis-dependent expression of Fas and Fas ligand (Fas-L) was detected by treatment with 2CdA. The proteolytic processing of procaspases-8 and -3 to produce active fragments, caspases-8 (p18) and -3 (p17), respectively, was observed after treatment with 2CdA, and suppressed by cycloheximide. Increases in the activities of caspases-8 and -3 were observed after 2CdA treatment. Their activation was also dependent on protein synthesis. These results indicated that 2CdA-induced apoptosis was triggered by phosphorylation of 2CdA followed by the protein synthesis-dependent expression of Fas and Fas-L and activation of caspases-8 and -3. PMID- 10673749 TI - Improved retroviral transduction of hematopoietic progenitors by combining methods to enhance virus-cell interaction. AB - One of the factors required for successful retroviral transduction is contact between viral particles and target cells. We hypothesized that combining agents that improve virus-target cell interaction via different mechanisms will increase transduction efficiency. We examined the transduction efficiency of leukemic K562 cells, primary normal and chronic myelogenous leukemia CD34+ cells with the amphotropic retroviral vector, G1Na, packaged in PA317 by enumerating G418 resistant colonies in semisolid media. We evaluated the ability of the recombinant fibronectin fragment, CH296, cationic lipids, or a transwell flow through system, alone or in combination to improve retroviral transduction. Transduction of K562 cells improved 1.5 to two-fold with lipids or CH296, while their combination improved transduction 2.5-fold. Transduction of K562 cells in the transwell flow-through system improved transduction three-fold. Transduction of normal (NL) CD34+ CFC improved 10-fold with lipids and 20-fold with CH296. Lipid and CH296 had synergistic effects. The transwell flow-through system improved transduction of normal CD34+ CFC 30-fold. Finally, similar to what was seen for K562 cells, transduction of CML CFC improved two- to three-fold with either CH296 or lipids, whereas the combination had synergistic effects. We conclude that any physical means that enhances contact between viral particles and target cells improves transduction. Two such methods that have different action mechanisms have additive or synergistic effects on transduction. PMID- 10673750 TI - Rapid and reliable detection of N-ras mutations in acute lymphoblastic leukemia by melting curve analysis using LightCycler technology. AB - We applied a new strategy for the detection of N-ras gene mutations based on LightCycler technology. We designed two sets of amplimers and internal hybridization probes representing N-ras codons 12/13 and codon 61, respectively. Genomic DNAs from 134 childhood acute lymphoblastic leukemia (ALL) patients (83 common ALL, nine pre-pre-B ALL, 19 pre-B ALL, 23 T-ALL) were amplified, followed by the analysis of the melting temperatures of the PCR products on the LightCycler. PCR products exhibiting an abnormal melting characteristic were directly sequenced. Sequence analyses unravelled nucleotide substitutions at codon 12 in 10 patients, at codon 13 in three, and at codon 61 in one case. The incidence of N-rasmutations (10%) is compatible with previous reports. The LightCycler technology facilitates the rapid analysis of other genes exhibiting hot spot mutations in human malignancies. PMID- 10673751 TI - Potential of LightCycler technology for quantification of minimal residual disease in childhood acute lymphoblastic leukemia. AB - A certain quantity of residual leukemic cells at several time points during chemotherapy of childhood acute lymphoblastic leukemia (ALL) was proved to predict outcome. Future childhood ALL treatment will take minimal residual disease (MRD) into consideration for stratification aiming at an individualization of chemotherapeutic regimens. Recently, the first quantitative real-time PCR assay for MRD detection was described using T cell receptor and immunoglobulin gene rearrangements as clonal markers. Quantitative real-time PCR was performed with TaqMan technology. Here, we present for the first time the potential of LightCycler real-time PCR technology to quantify MRD. We compare and assess different approaches for real-time PCR quantification of leukemic cells, based either on clone-specific primers and general fluorescence detection with SYBR Green, TaqMan probe or hybridization probes, or based on general PCR amplification and clone-specific detection with hybridization probes. MRD quantification with LightCycler real-time PCR technology is a sensitive, specific and incomparably rapid method that needs no post-PCR handling, hence eliminating contamination risk and saving time. Working towards the establishment of MRD quantification in routine diagnostics and towards treatment strategies based on these results, LightCycler quantitative PCR seems to be a promising new technique that makes results immediately available for treatment decisions. PMID- 10673752 TI - Quantitation of minimal residual disease in acute promyelocytic leukemia patients with t(15;17) translocation using real-time RT-PCR. AB - We took advantage of a recently developed system allowing performance of real time quantitation of polymerase chain reaction to develop a quantitative method of measurement of PML-RARalpha transcripts which are hallmarks of acute promyelocytic leukemia (APL) with t(15;17) translocation. Indeed, although quantitation of minimal residual disease has proved to be useful in predicting clinical outcome in other leukemias such as chronic myeloid leukemia or acute lymphoblastic leukemia, no quantitative data have been provided in the case of APL. We present here a method for quantitation of the most frequent subtypes of t(15;17) transcripts (namely bcr1 and bcr3). One specific forward primer is used for each subtype in order to keep amplicon length under 200 bp. The expression of PML-RARalpha transcripts is normalized using the housekeeping porphobilinogen deaminase (PBGD) gene. This technique allows detection of 10 copies of PML RARalpha or PBGD plasmids, and quantitation was efficient up to 100 copies. One t(15;17)-positive NB4 cell could be detected among 106 HL60 cells, although quantitation was efficient up to one cell among 105. Repeatability and reproducibility of the method were satisfying as intra- and inter-assay variation coefficients were not higher than 15%. The efficiency of the method was finally tested in patient samples, showing a decrease of the PML-RARalpha copy number during therapy, and an increase at the time of relapse. PMID- 10673753 TI - Comparison of nested competitive RT-PCR and real-time RT-PCR for the detection and quantification of AML1/MTG8 fusion transcripts in t(8;21) positive acute myelogenous leukemia. AB - The chromosomal translocation t(8;21)(q22;q22) is one of the most frequent karyotypic aberrations in acute myeloid leukemia (AML) and results in a chimeric fusion transcript AML1/MTG8. Since AML1/MTG8 fusion transcripts remain detectable by RT-PCR in t(8;21) AML patients in long-term hematological remission, quantitative assessment of AML1/MTG8 transcripts is necessary for the monitoring of minimal residual disease (MRD) in these patients. Competitive RT-PCR and recently real-time RT-PCR are increasingly used for detection and quantification of leukemia specific fusion transcripts. For the direct comparison of both methods we cloned a 42 bp DNA fragment into the original AML1/MTG8 sequence. The resulting molecule was used as an internal competitor for our novel competitive nested RT-PCR for AML1/MTG8 and as an external standard for the generation of AML1/MTG8 standard curves in a real-time PCR assay. Using this standard molecule for both PCR techniques, we compared their sensitivity, linearity and reproducibility. Both methods were comparable with regard to all parameters tested irrespective of analyzing serial dilutions of plasmids, cell lines or samples from t(8;21) positive AML patients at different stages of the disease. Therefore, both techniques can be recommended for the monitoring of MRD in these particular AML patients. However, the automatization of the real-time PCR technique offers some technical advantages. PMID- 10673754 TI - Simple methods for the rapid exchange of flow cytometric data between remote centers. PMID- 10673755 TI - Granulocyte colony-stimulating factor receptor expression and 11q23/MLL genotype in childhood acute lymphoblastic leukemia developing during the first 18 months of life. PMID- 10673756 TI - Flow cytometric detection of B cell abnormal maturation in chronic myeloid leukemia. PMID- 10673758 TI - Pseudotumor cerebri after treatment of relapsed acute promyelocytic leukemia with arsenic trioxide. PMID- 10673757 TI - 3 x 14 mg/m2 idarubicin during induction: results of a pilot study in children with AML. PMID- 10673759 TI - Good response to cyclosporine therapy in patients with myelodysplastic syndromes having the HLA-DRB1*1501 allele. PMID- 10673760 TI - Molecular psychiatry at the millennium. PMID- 10673762 TI - News in brief PMID- 10673761 TI - Comparative proteome analysis. Tissue homogenate from normal human hippocampus subjected to two-dimensional gel electrophoresis and Coomassie blue protein staining. PMID- 10673763 TI - Beware the chopsticks gene. AB - Population stratification is a potential source of error in psychiatric genetics. New study designs and statistical methods can help guard against this problem. Molecular Psychiatry (2000) 5, 11-13. PMID- 10673764 TI - The role of stress in the pathophysiology of the dopaminergic system. AB - In this review, we will examine the most recent preclinical evidence in support of the fact that both acute and chronic stress may have a detrimental impact on the normal function of the dopaminergic system. In recent decades, the term stress has changed its meaning from that of a 'non-specific body response' to a 'monitoring system of internal and external cues'; that is a modality of reaction of the mammalian central nervous system (CNS) which is critical to the adaptation of the organism to its environment. Compelling results have demonstrated that the dopaminergic system is important not only for hedonic impact or reward learning but also, in a broader sense, for reactivity to perturbation in environmental conditions, for selective information processing, and for general emotional responses, which are essential functions in the ability (or failure) to cope with the external world. In this, stress directly influences several basic behaviors which are mediated by the dopaminergic system such as locomotor activity, sexual activity, appetite, and cross sensitization with drugs of abuse. Studies using rat lines which are genetically different in dopamine (DA) physiology, have shown that even small alterations in the birth procedure or early life stress events may contribute to the pathophysiology of psychiatric disorders-in particular those involving central DA dysfunction-and may cause depression or psychotic derangement in the offspring. Finally, the fact that the dopaminergic system after stress responds, preferentially, in the medial prefrontal cortex (MFC), is thought to serve, in humans, as a protection against positive psychotic symptoms, since the increased DA activity in the MFC suppresses limbic DA transmission. However, excessive MFC dopaminergic activity has a negative impact on the cognitive functions of primates, making them unable to select and process significant environmental stimuli. Thus it appears that a critical range of DA turnover is necessary for optimal cognitive functioning after stress, in the response of the CNS to ever-changing environmental demands. Molecular Psychiatry (2000) 5, 14-21. PMID- 10673765 TI - Psychiatric genetics: back to the future. AB - For the last decade or more geneticists have been predicting that advances in molecular genetics are going to revolutionize our understanding of psychiatric disorders and human behavior. However, with a few exceptions, these expectations have yet to be fulfilled. As the century draws to a close and we contemplate the prospect of the complete sequence of the human genome it seems timely to consider the state of the field and to consider carefully how it might advance, the problems to be faced and the resources required. Molecular Psychiatry (2000) 5, 22-31. PMID- 10673766 TI - The human serotonin transporter gene linked polymorphism (5-HTTLPR) shows ten novel allelic variants. AB - The serotonin transporter (5-HTT) gene is a promising candidate for introducing the heritability of interindividual variation in personality and the genetic susceptibility for various psychiatric diseases. Transcription of the gene is modulated by a common polymorphism in its upstream regulatory region (5-HTT gene linked polymorphic region: 5-HTTLPR). The 5-HTTLPR consists of variation of the repetitive sequence containing GC-rich, 20-23-bp-long repeat elements. A deletion/insertion in the 5-HTTLPR was first reported to create a short (S) allele and a long (L) allele (14- and 16-repeats, respectively). Three other kinds of alleles (18-, 19- and 20-repeats) in addition to the S and L alleles in 5-HTTLPR have been reported. In the present study, we examined the 5-HTTLPR polymorphism in detail and identified ten novel sequence variants, concluding that the alleles reported as S and L are divided into four and six kinds of allelic variant, respectively. Subsequently, we developed a method for genotyping. The total number of alleles (14-A, 14-B, 14-C, 14-D, 15, 16-A, 16-B, 16-C, 16-D, 16-E, 16-F, 19, 20 and 22) in the 5-HTTLPR was 14 in our populations (Japanese: n = 131; Caucasian: n = 74) in the present study. In addition, a significant ethnic difference between Japanese and Caucasian populations was observed for distributions of alleles and genotypes (P < 0.0001 and P < 0.0001, respectively). Our results suggest that the analyses of the 5-HTTLPR should be revised by genotyping with a more complete subdivision of alleles. Molecular Psychiatry (2000) 5, 32-38. PMID- 10673767 TI - Dopamine induces a biphasic modulation of hypothalamic ANF neurons: a ligand concentration-dependent effect involving D5 and D2 receptor interaction. AB - Increasing evidence now suggests that more than one subtype of dopamine receptors is co-expressed in some of the central neurons. The neurobiological effects on the host cells when these receptors are concurrently activated by their common physiological ligand, dopamine, however, remains elusive. Among the members of the family of dopamine receptors, coupling of D1-like dopamine receptors to Gs and D2-like receptors to Gi proteins are known to augment or suppress cellular functions respectively, through modulation of adenylyl cyclase activity and consequently cAMP generation. Simultaneous activation of D1 and D2 receptors in transfected cell lines expressing the two cloned receptors, however, produced antagonistic effects. This is in contrast to in vivo studies, in which concurrent activation of D1-like and D2-like receptors by their respective agonists may induce synergistic or antagonistic effects or both. We report here that in long term rat hypothalamic cell cultures, activation of both D1-like (D1 and D5) and D2 receptors on atrial natriuretic factor-producing neurons by dopamine yields a biphasic response. The response is ligand concentration-dependent and involves type II adenylyl cyclases. This process is mediated primarily through antagonistic and synergistic interactions of D5 and D2 receptors as the event is mimicked by the concurrent activation of these two receptors co-transfected in CHO cells. Our present findings suggest a novel action of dopamine, and the biochemical processes involved may underlie some of the pharmacological actions of atypical anti-psychotic drugs. Molecular Psychiatry (2000) 5, 39-48. PMID- 10673768 TI - Identification and analysis of new sequence variants in the human tryptophan hydroxylase (TpH) gene. AB - The tryptophan hydroxylase (TpH) gene codes for the rate-limiting enzyme in serotonin biosynthesis. It is one of the major candidate genes for psychiatric and behavioral disorders. A polymorphism in TpH intron 7 has been shown to be associated with suicidal attempts, aggressive behavior and psychiatric illnesses. By systematically screening the TpH genomic sequence, we identified and confirmed an earlier report of four variants in the promoter region and localized six new sequence variants, ie two in intron 1b, one in exon 1c, one in intron 8, one in intron 9 and a microsatellite in the 3' region, 5687 bp downstream of the last exon 11. We analyzed these polymorphisms, as well as the one in intron 7, by Single Strand Conformation Analysis, microsatellite or restriction analysis in a collection of 175 West European Caucasian healthy subjects. The four variants in the promoter region are in complete linkage disequilibrium (frequencies of G-T-G T and T-C-A-G haplotypes are 0. 41 and 0.59, respectively). Deletion of GTT in intron 1b is rare (0. 7%) and so not informative. The rarer allele T of intron 1b polymorphism T3792A has a frequency of 0.34 and is in partial linkage disequilibrium with the more common alleles of intron 7, 8 and 9. The polymorphisms of these three introns are in complete linkage disequilibrium and the frequencies of haplotypes A-T-C and C-C-T are 0.36 and 0.64 respectively. We detected 10 different alleles in the microsatellite localized in the 3' region; allele '194' is in partial linkage disequilibrium with haplotype A-T-C of introns 7, 8, and 9. Analysis of these different polymorphisms will constitute an important tool for future studies between the TpH gene and psychiatric disorders. Molecular Psychiatry (2000) 5, 49-55. PMID- 10673769 TI - A haplotype at the DBH locus, associated with low plasma dopamine beta hydroxylase activity, also associates with cocaine-induced paranoia. AB - Low levels of dopamine beta-hydroxylase (DbetaH) protein in the plasma or cerebrospinal fluid (CSF) are associated with greater vulnerability to positive psychotic symptoms in several psychiatric disorders. DbetaH level is a stable, genetically controlled trait. DBH, the locus encoding DbetaH protein, is the major quantitative trait locus controlling plasma and CSF DbetaH levels. We therefore hypothesized that DBH variants or haplotypes, associated with low levels of DbetaH in the plasma, would also associate with greater vulnerability to cocaine-induced paranoia. To test this hypothesis, we first showed that a di allelic variant, DBH*5'-ins/del, located approximately 3 kb 5' to the DBH transcriptional start site, significantly associates with plasma DbetaH activity in European-Americans (n = 66). Linkage disequilibrium analysis of that polymorphism and DBH*444g/a, another di-allelic variant associated with DbetaH levels, demonstrated that alleles of similar association to DbetaH levels are in positive disequilibrium. We then estimated DBH haplotype frequencies in cocaine dependent European Americans rated for cocaine-induced paranoia (n = 45). As predicted, the low-DbetaH-associated haplotype, Del-a, was significantly more frequent (P = 0.0003) in subjects endorsing cocaine-induced paranoia (n = 29) than in those denying it (n = 16). Comparison to control haplotype frequencies (n = 145 healthy European-Americans) showed that the association predominantly reflected under-representation of Del-a haplotypes in those denying cocaine induced paranoia. We conclude that: (a) the two DBH polymorphisms we studied are associated with plasma DBH levels; (b) those two polymorphisms are in significant linkage disequilibrium in European Americans, with alleles of similar association to DbetaH levels in positive disequilibrium; and (c) the haplotype associated with low DBH activity is also associated with cocaine-induced paranoia. Molecular Psychiatry (2000) 5, 56-63. PMID- 10673770 TI - Identification of a polymorphism in the promoter region of DRD4 associated with the human novelty seeking personality trait. AB - Polymorphism in the human dopamine D4 receptor gene (DRD4) exon III has been associated in some but not all studies of the human personality trait of Novelty Seeking. We searched for polymorphisms in the 5' region of DRD4 and identified six polymorphisms as follows: -1217G Ins/Del, -809G/A, -616C/G, -603T Ins/Del, 602(G)8-9, and -521C/T. Associations between these polymorphisms and personality traits measured by the Temperament and Character Inventory (TCI) were investigated in 86 healthy Japanese volunteers. The -521C/T polymorphism was significantly associated with Novelty Seeking (P = 0.0001). Subjects with the C/C genotype exhibited the highest Novelty Seeking scores and those with the T/T genotype exhibited the lowest. A transient expression method revealed that the T variant of the C-521T polymorphism reduces transcriptional efficiency. The present study suggests a contribution of dopamine D4 receptor availability to individual differences in Novelty Seeking behavior. Molecular Psychiatry (2000) 5, 64-69. PMID- 10673771 TI - Antipsychotic drug-induced obesity in rats: correlation between leptin, insulin and body weight during sulpiride treatment. AB - Sulpiride (SUL, 20 mg kg-1 day-1) induces weight gain, hyperphagia, hyperprolactinemia, hypogonadism, and perhaps increased insulin sensitivity in rats. Leptin seems to signal the brain about the size of body fat stores and nutrient metabolism. We evaluated the basal serum leptin levels in rats after acute (1 h) or prolonged SUL or vehicle administration (10, 20 and 30 days). At days 10 and 30 leptin was also assessed during a glucose overload test. As the maximal weight gain during SUL administration is observed at days 10-15 of treatment, leptin was measured in a comparison group of insulin-treated rats (5 IU day-1 for 10 days). SUL-treated rats significantly gained weight. However, leptin levels were not significantly increased at any time-point of treatment. SUL did not affect insulin levels either. By contrast, leptin levels were significantly elevated after insulin administration, along with weight gain and hyperinsulinemia. An opposite correlation was also observed at day 10: leptin and insulin correlated negatively in the SUL group and positively in the insulin group. In addition, leptin and the magnitude of weight gain tended to correlate positively after SUL treatment, but negatively after insulin administration. SUL treated rats, thus, appear to exhibit an unusual type of weight gain, characterized by normal circulating leptin and insulin levels. Such a particular leptin profile may be related to hyperprolactinemia, hypogonadism or lack of hyperinsulinemia. Molecular Psychiatry (2000) 5, 70-76. PMID- 10673772 TI - Family-based linkage disequilibrium mapping using SNP marker haplotypes: application to a potential locus for schizophrenia at chromosome 22q11. AB - Family-based linkage disequilibrium mapping using SNP markers is expected to be a major route to the identification of susceptibility alleles for complex diseases. However there are a number of methodological issues yet to be resolved, including the handling of extended haplotype data and analysis of haplotype transmission in sib-pair or family trio samples. In the present study, we have analysed two dinucleotide repeat and six SNP markers at the COMT locus at chromosome 22q11, a region implicated in psychosis, for transmission distortion in 198 Chinese schizophrenic family trios. When individual markers were analysed using the TDT, two showed modest evidence of transmission distortion (186C/T, P = 0.04; Val158Met, P = 0.01). Using haplotypes of paired markers analysed by the program TRANSMIT, the most significant P value was 0.001, for the Met158Val and 900ins/delC polymorphisms in the COMT gene. The global P value for the haplotypes of all six SNP markers tested was 0.004, largely a result of the excess transmission of two extended haplotypes which differed at the marker 408C/G. The exclusion of this marker from the analysis gave a global P value of 0.002 and produced a five marker haplotype system which was significant at P = 0.0006. This haplotype consisted of the alleles -287G:186C:Val158:900insC:ARVCF930C, which may represent a background haplotype for the transmission of a schizophrenia susceptibility allele at chromosome 22q11. Our results support the hypotheses that either COMT is itself a susceptibility gene, or more likely that this region of chromosome 22 contains a susceptibility gene that is in linkage disequilibrium with COMT alleles. Molecular Psychiatry (2000) 5, 77-84. PMID- 10673773 TI - Comparative proteome analysis of the hippocampus implicates chromosome 6q in schizophrenia. AB - Comparative brain proteome analysis is a new strategy to discover proteins and therefore genes whose altered expression may underlie schizophrenia. This strategy does not require an a priori theory of the pathogenesis or the mode of inheritance of schizophrenia. Using proteome analysis we previously compared the hippocampal proteome, that is, those proteins expressed by the hippocampal genome, of seven schizophrenic individuals with the hippocampal proteome of seven control individuals, matched for age and post mortem delay.1 We found 18 proteins that were significantly altered in concentration in the schizophrenic hippocampus (P < 0.05), when compared to control tissue. One of these proteins was characterised, by N-terminal sequencing, as diazepam binding inhibitor whose gene maps to 6q12-q21. Here we characterise a further three of the 18 proteins as: manganese superoxide dismutase, 6q25.3, T-complex protein 1, 6q25.3-q26 and collapsin response mediator protein 2, 8p21. That three of these four characterised proteins should map to the long arm of the same chromosome is significant (P < 0.002) and suggests the importance of chromosome 6q in schizophrenia. These results indicate that antioxidant defence is altered in the schizophrenic hippocampus and suggest that segregation distortion, of schizophrenia susceptibility genes, may be a possible causative factor in the high incidence of schizophrenia. Molecular Psychiatry (2000) 5, 85-90. PMID- 10673774 TI - Serotonin transporter gene and schizophrenia: evidence for association/linkage disequilibrium in families with affected siblings. AB - The serotonergic (5-HT) system has been implicated in the etiopathogenesis of psychoses. Since the 5-HT transporter plays an important role in regulation of 5 HT transmission, its gene can be considered as a candidate for vulnerability to psychiatric disorders. Two polymorphic sites of the 5-HT transporter gene-5 HTTLPR, a VNTR in the 5' regulatory region, and a VNTR in the second intron-were studied in a sample of 61 families with schizophrenia for transmission disequilibrium. Each family contained at least two siblings affected with schizophrenia or schizoaffective disorder (mainly schizophrenic). One hundred and thirty-nine affected offspring with parental information for genotyping, were available for analysis. No preferential transmission of either short or long alleles of the promoter polymorphism was observed. However, a transmission distortion was detected for alleles of the intronic VNTR polymorphism (chi2TDT max =14.33; P = 0.0002; corrected P value = 0.0003) resulting in more frequent than expected transmission of the 12 repeat allele. This finding adds additional evidence to the idea that the serotonergic system may be involved in development of psychoses. Molecular Psychiatry (2000) 5, 91-95. PMID- 10673775 TI - Association between tridimensional personality questionnaire (TPQ) traits and three functional polymorphisms: dopamine receptor D4 (DRD4), serotonin transporter promoter region (5-HTTLPR) and catechol O-methyltransferase (COMT). AB - Dopamine D4 receptor (DRD4), serotonin transporter promoter regulatory region (5 HTTLPR) and catechol O-methyltransferase (COMT) polymorphisms were examined for association with TPQ personality factors in 455 subjects. Significant interactions were observed by multivariate analysis, (COMT x 5-HTTLPR: Hotelling's Trace = 2.3, P = 0.02) and by subsequent univariate 3-way ANOVA when Novelty Seeking (NS) was the dependent variable: 5-HTTLPR x D4DR (F = 6.18, P = 0.03) and COMT x 5-HTTLPR (F = 4.42, P = 0.03). In the absence of the short 5 HTTLPR allele and in the presence of the high enzyme activity COMT val/val genotype, NS scores are higher in the presence of the DRD4 seven-repeat allele. The effect of these three polymorphisms on NS was also examined using a within families design. Siblings who shared identical genotype groups for all three polymorphisms (COMT, DRD4 and 5-HTTLPR) had significantly correlated NS scores (intraclass coefficient = 0.39, F = 2.26, P = 0.008, n = 49) whereas sibs with dissimilar genotypes in at least one polymorphism showed no significant correlation for NS scores (intraclass coefficient = 0.177, F = 1.43, P = 0.09, n = 110). Similar interactions were also observed between these three polymorphisms and Novelty Seeking when the 150 independently recruited and non-related subjects were analyzed. The current results are consistent with two earlier reports in which we demonstrated an interaction between the 5-HTTLPR and DRD4 polymorphisms in 2-week-old neonates, in the same children assessed again at 2 months of age and in adults. Molecular Psychiatry (2000) 5, 96-100. PMID- 10673777 TI - Pathological gambling and DNA polymorphic markers at MAO-A and MAO-B genes. AB - This study was conducted to detect a possible association of MAOA and/or MAOB genes with pathological gambling (PG). DNA polymorphisms in MAOA and MAOB genes were screened by molecular analysis in 68 individuals (47 males and 21 females) meeting ICD-10 and DSM-IV criteria for pathological gambling and 68 healthy comparison controls matched for age and sex. There were no significant differences between pathological gamblers and healthy volunteers in overall allele distribution at the MAOA gene polymorphism. However there was a significant association between allele distribution and the subgroup of severe male gamblers (n = 31) compared to the males in the group of healthy volunteers (chi2 = 5246; df = 1; P < 0.05 [Bonferroni corrected]). No association was found between the MAOB polymorphic marker and PG. Allele variants at the MAOA, but not the MAOB gene may be a genetic liability factor in PG, at least in severe male gamblers. Molecular Psychiatry(2000) 5, 105-109. PMID- 10673776 TI - DRD4 exon III VNTR polymorphism-susceptibility factor for heroin dependence? Results of a case-control and a family-based association approach. AB - Dopaminergic abnormalities are implicated in the pathogenesis of substance abuse.1 Recently, two reports have been published suggesting an association between opioid dependence and presence of long alleles of the dopamine D4 receptor (DRD4) gene exon III VNTR.2, 3 We have attempted to replicate this finding using a two-tiered strategy employing independent case-control and family based association samples. Our study was possibly the largest candidate gene association study to date on opioid dependence in a sample of 815 subjects, 396 of whom were patients. We found long alleles of the DRD4 exon III VNTR in similar frequency among 285 heroin addicts and 197 controls. Furthermore, no preferential transmission of long alleles to affected offspring was observed in a sample of 111 patients and their parents. Our results, therefore, do not support the hypothesis that alleles of the DRD4 exon III VNTR are susceptibility factors for opioid dependence in man. Molecular Psychiatry(2000) 5, 101-104. PMID- 10673779 TI - Volume 6 and the new millennium. PMID- 10673778 TI - Modified structure of the human serotonin transporter promoter. AB - Recently, several studies have reported an association between anxiety traits, affective disorders and autism and alleles of a functional promoter polymorphism (5HTT-LPR) in the human serotonin transporter (5HTT, SERT).1-3 The mechanistic basis for allelic differences in transporter transcription are presently unknown. To explore this issue, we cloned the human 5HTT promoter region from a PAC genomic library and now describe an unreported 381-bp insert between the polymorphic region and the transcription start site. We verified the presence of this novel sequence by Southern hybridization of genomic digests and PCR amplifications from multiple unrelated individuals. Sequence analysis of the novel region reveals a number of canonical transcription factor binding sites (eg AP1, Elk1, NFkappaB) that may be important in controlling the response of the 5HTT gene to regulatory factors. PCR studies of genomic templates reveal a low level of amplification of a deleted template matching the size of the originally reported 5HTT promoter. This deleted template is absent from PAC amplifications, suggesting that the human 5HTT promoter may exhibit in vivo instability. Molecular Psychiatry (2000) 5, 110-115. PMID- 10673780 TI - Publisher's announcement. PMID- 10673781 TI - Isolation and identification of Candida from the oral cavity. AB - A number of methods of sampling the oral cavity for the presence of candida have been developed. Such techniques play an important role in the diagnosis and management of oral candidosis. In the past, identification of candida isolated from the oral cavity has usually been limited to the genus Candida or to the species C. albicans. However, with the recognition that Candida species differ in the production of putative virulence factors and sensitivity to antifungal agents, greater emphasis has been placed on identification of isolates to species level. As a result a range of commercially available systems for yeast identification can now be used in conjunction with traditional identification procedures. PMID- 10673782 TI - Influence of (S)-ketoprofen and fluoride on caries in rats. AB - OBJECTIVE: Non-steroidal anti-inflammatory agents (e.g., ketoprofen) used topically appear to be effective in reducing bone loss in the ligature model of periodontitis. Ketoprofen, in common with some food preservatives, e.g., benzoate and sorbate, is a weak acid. Fluoride, too, may behave as a weak acid and, similar to the other agents, may exert antibacterial effects. The purpose of the present study was to determine whether a combination of (S)-ketoprofen, an enantiomer of ketoprofen, alone or in combination with fluoride, would suppress Streptococcus sobrinus populations and reduce the incidence of dental caries in rats. MATERIALS AND METHODS: Toothpastes containing ketoprofen and/or monofluorophosphate were applied to the teeth of six groups of 20 rats twice daily for 5 weeks. RESULTS: Fewest S. sobrinus were found in the group treated with a paste containing 3% (S)-ketoprofen + 0.1% F. This group also displayed the lowest incidence of smooth surface caries of all groups. Severity of sulcal surface caries was also lowest in this group. CONCLUSIONS: Results from this study show that the (S) enantiomer of ketoprofen enhances the caries protective effect of fluoride. It is conceivable that this combination could be effective in combating the two most common maladies of the mouth; periodontal disease and dental caries. PMID- 10673783 TI - Long-term follow-up of patients treated with acupuncture for xerostomia and the influence of additional treatment. AB - OBJECTIVE: To determine the long-term effects of acupuncture in patients with xerostomia of different etiologies and the influence of additional treatment. DESIGN: Retrospective study. SUBJECTS: Seventy patients, between the ages of 33 and 82, with xerostomia due to primary and secondary Sjogren's syndrome, irradiation and other causes were included. The median duration of xerostomia was 32 months. METHODS: Salivary flow rates (SFR) for whole unstimulated and stimulated saliva were used as indicators of effects of treatment. Data from 67/70 patients were analyzed 6 months following a baseline course of 24 acupuncture treatments using two-way ANOVA. Patients data up to 3 years were also compared by those who chose to receive additional acupuncture treatment vs those who did not. These data were analyzed descriptively. RESULTS: Statistically significant differences in unstimulated and stimulated salivary flow rates (P < 0.01) were found in all etiological groups after 24 acupuncture treatments and up to 6 months follow-up compared to baseline. Three years observation of these patients showed that patients receiving additional acupuncture treatment had a consistently higher median SFR in both unstimulated and stimulated saliva compared to patients who chose not to continue acupuncture. The upper limits of the interquartile range were also higher. CONCLUSIONS: This study shows that acupuncture treatment results in statistically significant improvements in SFR in patients with xerostomia up to 6 months. It suggests that additional acupuncture therapy can maintain this improvement in SFR for up to 3 years. PMID- 10673784 TI - Oral tuberculosis: a clinical evaluation of 42 cases. AB - OBJECTIVES: A retrospective review of a large series of oro-facial cases of tuberculosis to analyse clinical, histopathological, and radiological aspects, as well as those of chemotherapy. MATERIALS AND METHODS: A total of 42 cases of tuberculosis of the oro-facial region were examined. Thirteen patients had a primary form and 29 a secondary form of the disease. Diagnosis was based on careful clinical examination, Mantoux reaction, histopathological examination, microbiological cultures and immunological investigation with the detection of antibodies against Mycobacteria in the patients' serum (ELISA). RESULTS: Cases examined consisted of 27 males and 15 females. The age range was 3 to 73 years (mean age 31 years). Clinical manifestations comprised oral ulcers in 69.1%, bone involvement in 21.4%, and salivary gland and/or lymph node involvement in 14.3%. A total of 79.4% patients with secondary disease had pulmonary lesions, 15 of whom showed clinical and radiological signs of activity; there was one case of bilateral renal lesions and two of skin lesions. CONCLUSIONS: Oro-facial tuberculosis is often difficult to diagnose and it should be an important consideration in the differential diagnosis of lesions that appear in the oral cavity. The most important diagnostic tools remain a careful clinical evaluation, biopsy for histologic study, as well as acid-fast stains, culture, and immunological assays, and skin testing. PMID- 10673785 TI - Immunohistochemical detection of insulin-like growth factor-I in the labial salivary glands of patients with Sjogren's syndrome. AB - OBJECTIVE: The purpose of the present study was to investigate the presence of insulin-like growth factor-I (IGF-I) in the labial salivary glands of patients with Sjogren's syndrome and healthy controls and to determine if there are any differences between these two groups. DESIGN: An immunohistochemical study. SUBJECTS AND METHODS: Twenty-five patients with Sjogren's syndrome, 20 healthy controls and 20 patients with mucoceles of the lip were used in this study. All individuals underwent a systemic evaluation and a lip biopsy. Sections from the lip biopsies were stained with haematoxylin and eosin (H&E). Immunohistochemical staining was also performed using a three-step indirect immunoperoxidase for IGF I. RESULTS: The light microscopic examination revealed the presence of a mononuclear infiltration in the labial salivary glands of patients with Sjogren's syndrome. Most of the infiltrates were lymphocytes. Immunohistochemically an intense staining result was apparent in the same group. In contrast sections of labial salivary glands of healthy individuals and of patients with mucoceles revealed very weak staining. CONCLUSIONS: The above findings and the fact that both lymphocytic infiltration and IGF-I were predominantly seen in ductal regions, suggest that IGF-I may be a target of autoimmunity in Sjogren's syndrome. PMID- 10673786 TI - Evaluation of p53, PCNA, Ki-67, MDM2 and AgNOR in oral peripheral and central giant cell lesions. AB - OBJECTIVES: Peripheral giant cell lesion (PGCL) and central giant cell lesion (CGCL) of the jaws have a distinct clinical behaviour. Whether such biological differences are supported by a distinct pattern of proliferation markers or cell cycle associated proteins expression is not known. Therefore the purpose of the present study was to compare the immunohistochemical expression of p53, MDM2, Ki 67, PCNA and the histochemical expression of argyrophilic nuclear organiser region (AgNOR) on PGCL and CGCL of the jaws. MATERIALS AND METHODS: Paraffin wax blocks of 14 cases of PGCL and 12 cases of CGCL were retrieved. A biotin streptavidin amplified system was used for identification of the antigens. The AgNOR number was also evaluated. RESULTS: Ki-67 immunoreactivity was greater in the mononuclear cells of PGCL compared to CGCL. PCNA and AgNOR staining were similar in PGCL and CGCL. Prominent MDM2 immunoreactivity was observed in all tissues investigated. By contrast, there was no p53 immunoreactivity. CONCLUSIONS: Although CGCL present a more aggressive clinical behaviour, it has a decreased proliferative activity compared to PGCL. Finally, p53, MDM2, PCNA, Ki 67 immunohistochemical expression and AgNOR histochemical expression do not reflect their distinct biological behaviour. PMID- 10673787 TI - Human immunodeficiency virus-positive individuals with oral hairy leukoplakia are able to mount cytotoxic T lymphocyte responses to Epstein-Barr virus. AB - OBJECTIVE: Oral hairy leukoplakia (OHL) is a white lesion of the tongue that is caused by Epstein-Barr virus (EBV) and occurs mainly in people infected with human immunodeficiency virus (HIV). The aim of this study was to determine whether the presence of OHL reflects the absence of EBV-specific cytotoxic T lymphocyte (CTL) activity. SUBJECTS AND METHODS: EBV-specific CTL responses were measured in HIV-positive homosexual men with OHL, HIV-positive homosexual men without OHL, and HIV-negative homosexual men. Also, the phenotypes of cells responsible for EBV-specific responses were studied. RESULTS: Eighty percent (8/10) of HIV-positive subjects with OHL, 52% (12/23) of HIV-positive subjects without OHL, and 83% (15/18) HIV-negative subjects had a positive anti-EBV CTL response (P = 0.004, Kruskal-Wallis test). Two HIV-positive subjects showed a greater anti-EBV CTL response after developing OHL than before the appearance of OHL Additional experiments showed that CD8-positive T cells and CD4-positive T cells were responsible for the EBV-specific CTL responses. CONCLUSION: Our data show more EBV-specific CTL activities in HIV-positive individuals with OHL than in HIV-positive individuals without OHL. Whether the presence of EBV-specific CTL contributes to resolution of OHL remains to be clarified. PMID- 10673788 TI - [Urinary incontinence after radical prostatectomy: severe ultrasonography changes following periurethral injection of polytetrafluoroethylene]. AB - The periurethral injection of polytetrafluoroethylene in the treatment of stress urinary incontinence is a therapeutic option introduced more than 30 years ago which is considered rapid, effective and burdened with low complications. This particular therapeutic option when utilized for the treatment of post prostatectomy incontinence, presents some problems due to the oncologic follow-up of these patients with particular regard to the presence of local recurrences. In fact these injections should not interfere with the standard methods of evaluation of the urethro-vescical anastomosis (DRE and TRUS). The polytetrafluoroethylene in our experience does not accomplish with these requirements because it modifies the palpatory and sonographic findings hindering a reliable evaluation of that region. PMID- 10673789 TI - [Transurethral treatment of bladder diverticuli]. AB - Since October 1994 we performed endoscopic treatment of bladder diverticula, in combination with the resolution of cervico-urethral obstruction, in 12 patients. In a first group of five patients we used the incision of the diverticular neck, while in a second group of seven patients we associated the electrocoagulation of the diverticular mucosa to the opening of the diverticular neck. The better results have been obtained in the second group of patients in which the diverticula had markedly shrunk in two cases and had totally disappeared in five. Transurethral treatment of bladder diverticula, with incision of the diverticular neck and the fulguration of the diverticular mucosa, combined with transurethral operations for removing the obstruction of the lower urinary tract, is a safe, fast and effective method. PMID- 10673790 TI - [Role of urinary cytology in the study of bladder cancer. Comparison with the BTA test]. AB - Early diagnosis of bladder neoplasia at the moment makes use of urinary cytology and cystoscopy. The authors describe the results of a study on 62 patients (56 men and 6 women) with bladder neoplasia, and compare the results of urinary cytology to the BTA test (Bladder Tumor Antigen test), ones, after considering histological results of TURB (Trans Urethral Resection Bladder) or cystectomy. Sensibility of urinary cytology was quite better than BTA test sensibility (80.6% vs 48%). Finally it's described a study on 450 cases of istologically controlled bladder carcinomas, on which had been executed a cytological analysis before surgical treatment. The outcome of cytological diagnosis on 414 cases (92%) was neoplasia. On the basis of their experience, the authors regard cytology as a fundamental method in diagnostic iter and in follow-up of patients with bladder neoplasia. PMID- 10673791 TI - Terazosine and tamsulosin in non bacterial prostatitis: a randomized placebo controlled study. AB - Eighteen patients with inflammatory process of the prostate met criteria for the inclusion in the study: 1) non bacterial prostatitis; 2) no previous treatment. Then they were randomized into three groups as it follows: terazosine, tamsulosin and placebo. Alpha-blockers and placebo were given for two months, after which further uroflowmetry was performed. Symptom score was evaluated before and after treatment. Terazosine was effective in reducing TO (p = 0.01) as tamsulosin and placebo did not. Both terazosine (p = 0.034) and tamsulosin (p = 0.006) reduced max TQ as placebo did not. Symptom score significantly improved in patients receiving terazosine (p = 0.0002) and tamsulosin (p = 0.001) while insignificantly in whose receiving placebo. PMID- 10673792 TI - A multivariate analysis of lower urinary tract ageing and urinary symptoms: the role of fibrosis. AB - An investigative trial including 72 patients who underwent open surgery for benign prostatic hypertrophy (BPH) induced urinary symptoms was carried out with the purpose to obtain a deeper insight in the pathophysiology of this clinical picture. Prostate weight, stroma to parenchyma ratio, bladder wall fibrosis, I PSS score, residual urine and uroflow obtained from these patients were processed by statistical multivariate analysis. The results point out the pivotal impact of prostate and bladder wall fibrosis in conditioning biological and chronological ageing of the lower urinary tract and relative symptoms. PMID- 10673793 TI - Antiandrogens: a summary review of pharmacodynamic properties and tolerability in prostate cancer therapy. AB - This article provides a summary of the pharmacodynamic properties of major antiandrogens as well as an extensive review of their tolerability. Presently there are two classes of androgen receptor antagonists: the so-called pure, non steroidal antiandrogens which include flutamide, nilutamide and the more recent bicalutamide and the steroidal antiandrogens cyproterone acetate, megestrol acetate and WIN 49596. Although non steroidal and steroidal compounds have been found to be equally effective in the treatment of prostate cancer presently no studies comparing the use of steroidal or non steroidal antiandrogens with chemical or surgical castration have evaluated quality of life per se. The only advantage of cyproterone acetate on pure antiandrogens seems to be the low incidence of hot flushes; a commonly reported adverse effect of androgen ablative therapy. However, hepatotoxicity associated with long term daily doses of 300 mg daily and the unacceptably high incidence of cardiovascular side effects (10%) should restrict its use to patients who are intolerant of pure antiandrogen compound. In contrast to steroidal compound nonsteroidal compounds let sexual potency to be retained, which is an important consideration with respect to the quality of life of some patients and, at present, the main indication for monotherapy with the pure antiandrogens. As regard as pure antiandrogens clinically important adverse events including gastrointestinal events, particularly diarrhea and occasional disturbances of liver function related to flutamide treatment and antabuse effect, problems with light-dark adaptation and rare interstitial pneumonitis related to nilutamide indicates the bicalutamide, due to its better tolerability profile, together with its once-daily oral administration regimen, could be considered the antiandrogen of first choice in the treatment of prostatic cancer. PMID- 10673794 TI - [Leydig cell tumor of the testis. Report of 3 clinical cases]. AB - Leydig cell tumours account for only 1-3% of all testicular neoplasms. They are commonly detected as palpable masses, but can present with precocious puberty, gynecomastia or infertility. Although they are usually benign, about 10% of the reported cases have evolved as a malignant neoplasm. There are no histologic features pathognomonic for malignancy and metastasis is the only certain criterion of a malignant. Leydig cell tumour. The follow-up must be prolonged. The treatment of this tumour is orchidofuniculectomy by inguinal approach, associated with retroperitoneal lymphadenectomy when regional nodes are involved. We reports our experience on three cases of Leydig cell tumour of the testis treated with orchidofuniculectomy. The patients have been submitted to a 9 months, 11 months and 10 years follow-up respectively, and at present they are without signs of disease. PMID- 10673795 TI - Renal leiomyosarcoma: report of three cases and review of the literature. AB - In the present study three cases of renal leiomyosarcoma are presented. All the patients were complaint about pain at right or left lumbar area and microscopic or gross haematuria. The clinical examination did not confirm any pathologic finding. The ultrasonographic and CT scan investigation of the patients revealed a renal mass. Under the frozen section diagnosis of malignant tumor in two cases and the cytological diagnosis of renal cell carcinoma in the third case, a radical nephrectomy was performed. In all the cases the pathological diagnosis was renal leiomyosarcoma. Because of the rarity of this neoplasm, the literature is reviewed, presenting the symptoms, the radiological findings, the diagnostic criteria and the differential diagnosis of the tumor. Radical nephrectomy remains the treatment of choice for this tumor, which exhibits an aggressive biological behavior and an unfavorable prognosis. PMID- 10673796 TI - [Interstitial cystitis: epidemiology]. AB - The epidemiological assessment of intestitial cystitis (IC) is not definitive as no diagnostic criteria, such as endoscopy or biochemical and anatomopathological examination, exist. The diagnosis is solely based on symptoms like urgency, frequency and pelvic pain. The first studies on the population date back from 20 years ago and show a percentage of 10 cases every 100 thousand inhabitants. There is weak link between genetic factors, immunological diseases, previous cystitis or eating habits and intestitial cystitis. Epidemiological studies have highlight the frequency of this disease, and stressed the importance of stricted behavioural rules for the first stages of intestitial cystitis. PMID- 10673797 TI - [Urgency-frequency syndrome in women: interstitial cystitis and correlated syndromes]. AB - In our clinical practice we encountered urgency-frequency syndrome in female patients. Only in the 3.6% is possible to diagnose a typical interstitial cystitis (IC). In the 63.6% we observed only local trigonal squamous metaplasia (leucoplasia), it could be considered a paraphysiological condition present in 50 70% of fertile women, its rigid, not impermeable epithelium may offer an aethiological hypotesis for the dysuric syndrome. In the treatment of this lesion by endoscopic infiltration we had syntomatological results with 47.8% of patients even if only for a short period (one-two years). This treatment is simple and can be repeated, if the patient is responsive. We noticed that the results did not change even if we used different drugs probably due to the role of a physical detachment of leucoplasia from bladder trigon. PMID- 10673798 TI - [Interstitial cystitis: medical treatment]. AB - Interstitial cystitis is a chronic disease of the bladder and its etiology is still unknown. In this article we treat the different approaches to this disease, supported by the fact that a clear therapeutic protocol doesn't exist. PMID- 10673799 TI - [Interstitial cystitis: surgical treatment]. AB - Surgical therapy of interstitial cystitis must never be considered a first option but must be reserved for cases, less than 10%, in which conservative therapy has proven ineffectual. Surgical therapy includes a variety that started at the turn of the century. Neurosurgical denervation and perivesical denervation like cysto cystoplasty and cystolysis, manipulate the innervation to reduce the bladder's hypersensitivity. This surgical approach may be considered in patients in whom bladder capacity is normal. The results are uncertain and the complications like neurogenic bladder relevant. Enterocystoplasty is much more widespread because interstitial cystitis is a benign disease that rarely required radical surgery. Augmentation cystoplasty and substitution cystoplasty are two variants but only the later has a rationale as it involves the resection of the detrusor which is the source of the pain. Detubularization drastically reduced urinary incontinence. The resection of the detrusor can be supratrigonal, subtrigonal or at the proximal urethra like in the orthotopic neobladder. If urinary diversion is chosen, the bladder must be removed. Before recommending surgical therapy each patient should undergo tests for the localization of the pain; moreover psychological and gynaecological evaluations should be made. If the bladder capacity exceeds 400 cc surgical operation is not advisable. If, on the other hand, the bladder capacity is lower than 400 cc substitution cystoplasty is first choice. If the patient suffers from trigonal cystitis or urethral hypersensitivity, urinary diversion is a better therapy. According to the questionnaires send to the Urologic Departments in Lombardy in 1998, the most widespread type of operation seems to be supratrigonal cystectomy + enterocystoplasty and augmentation cystoplasty. Subtrigonal cystectomy or urinary diversion are only occasionally chosen; continent pouch is the least frequent therapy at all. PMID- 10673800 TI - [Interstitial cystitis: local electromotive drug administration (EMDA)]. PMID- 10673801 TI - Storytelling. A strategy for living and coping with cancer. AB - The purpose of this focused program evaluation was to explore attitudes and beliefs about storytelling as a strategy for coping with cancer among participants who attended a cancer-related storytelling workshop. The response rate was 70% (n = 94) and included persons with a diagnosis of cancer, their loved ones, and members of the public. The program coordinators used a theoretical model described by Heiney (1995) that explains how storytelling may produce therapeutic effects in four domains: cognitive, affective, interpersonal, and personal. A questionnaire was designed to determine the extent that conference participants perceived therapeutic benefits in these domains as a result of attending the workshop. Statistical analysis consisted of descriptive summaries of individual questions and domain scores. Findings showed that 97% of the respondents agreed that storytelling was a helpful way to cope with cancer. Most of the respondents reported agreement with the therapeutic benefits of storytelling in all domains, with 85% agreeing that hearing others' stories of living with cancer gave them hope. Although the results of the evaluation were very positive, further study is needed to demonstrate the efficacy of storytelling as a strategy for coping with cancer. PMID- 10673802 TI - The meaning of "not giving in". Lived experiences among women with breast cancer. AB - This article explores the meaning to women with breast cancer of "not giving in". Giorgi's phenomenological method was applied, and data were collected through open interviews. Ten women with breast cancer participated. The analysis resulted in a general structure of the phenomenon studied, including six key constituents: accepting the challenge to go on living, working actively on the healing process, finding something important to live for, gaining insights about life itself, experiencing awareness and avoidance, and introducing radical change in life. The results are consistent with literature about strategies in facing death and development as human conditions. Understanding the phenomenon of "not giving in" seems to be crucial for nurses in helping women with breast cancer to mobilize the inner power to survive and develop as human beings. PMID- 10673803 TI - An analysis of the concept of risk. AB - The meaning of the word "risk" has changed throughout history. Once a neutral term, risk has come to represent a combination of probability and something adverse or dangerous. Phenomena that were previously referred to as hazards, dangers, or uncertainties are now labeled as risks. Although risk touches every aspect of health and human welfare, the dimensions of risk as conceptualized in the fields of epidemiology, nursing science, medical science, and lay health are qualitatively different. Risk has not been examined as a concept in nursing literature or research, although risk and related terms are defined in a few nursing textbooks. Using the evolutionary method of concept analysis, risk is examined as a concept. This analysis was undertaken to define and clarify the concept and dimensions of risk as they relate to risk for disease. A sound understanding of risk as a concept is critical for developing an empirical knowledge base in nursing and directing nursing research examining issues related to risk for developing diseases such as cancer. PMID- 10673804 TI - Nurse-patient communication in cancer care. A review of the literature. AB - Patients with cancer seem to experience distress particularly in the first period after diagnosis, and are likely to develop an affective disorder in the first 2 to 3 months. Communicative behaviors of nurses seem to play an important role in meeting the cognitive and affective needs of patients with cancer. This review of the literature examines the communicative behaviors of nurses during care activities with patients who have cancer. The studies show that emphasis is placed on the affective side, in which facilitating behaviors such as empathy, touch, comforting, and supporting are considered essential in caring for patients with cancer. Unfortunately, further studies in this review demonstrate that communication in oncologic care is complicated by such emotionally laden issues as the consequences associated with the life-threatening character of the disease and the far-reaching consequences of the medical treatment. This results in barriers to effective communication between patients with cancer and nurses. It is important, therefore, that nurses working with patients who have cancer are provided both structurally and repeatedly with continuing education programs in communication. Finally, most of the studies covered in this review have an explorative character. Future research in this area should pay attention to the use of controlled studies, large sample sizes, and observational instruments. PMID- 10673805 TI - Perceptions of caring among patients with cancer and their staff. Differences and disagreements. AB - The current dyadic study investigated (a) patient and staff perceptions of the importance of caring behaviors, patient health, quality of life, and greatest health-related concern; (b) patient anxiety and depression (Hospital Anxiety and Depression Scale); and (c) staff views of patient perceptions of the importance of caring behaviors. The study included 21 matched patient-staff dyads. Three questionnaire versions of the Caring Assessment Instrument were used to tap patient (CARE-P) and staff (CARE-S) perceptions, and staff views of patient perceptions (CARE-SP). There were no correlations between patient and staff perceptions of the importance of caring behaviors, patient health, quality of life, or greatest health-related concern. However, staff views of patient perceptions about the importance of caring behaviors were strongly correlated with their own perceptions. Staff ratings of the importance of caring behaviors were not related to patient anxiety, depression, health, and/or quality of life. Patient depression was negatively correlated with three CARE-Q subscales. The results indicate that staff are not successful in judging the importance of caring behaviors, health, quality of life, and greatest health-related concern for individual patients. The major implication is that staff must be open to patient perceptions of what caring behaviors are important, and must validate their own perceptions of patient needs and concerns. PMID- 10673806 TI - Implementation of an oral care standard for leukemia and transplantation patients. AB - The purpose of this project was to develop an oral care standard on two nursing units in a university hospital where care was given to patients undergoing bone marrow or stem cell transplantation (BMSCT) and other treatments for leukemia. Strategies used in this interdisciplinary effort included collaboration, consultation, education, and evaluation. In the collaboration phase, a core group of nurses talked with staff about current practices, reviewed literature and published standards, examined protocols from other institutions, decided on goals, and developed the standard. Consultation with a dentist, pharmacist, and physician occurred before completion of the standard. The education phase included in-service sessions for nurses and technicians. The evaluation phase, which occurred in two phases, focused on checking to see if the goals had been met, including tolerability and adherence. The first phase allowed identification of problem areas and subsequent revisions, whereas the second phase evaluated adherence at a later time point. Overall, most of the patients adhered to the standard. Future implications include specific recommendations such as an emphasis on oral care, documentation, and patient and staff education. This project is an example of how nurses addressed the challenge of implementing an acceptable oral care standard to decrease patients' oral complications and distress. PMID- 10673807 TI - An instrument to measure symptom experience. Symptom occurrence and symptom distress. AB - This article describes the development of an instrument that measures symptom experience (symptom occurrence and symptom distress). The Adapted Symptom Distress Scale-2 (ASDS-2), adapted from the McCorkle and Young Distress Scale, is a 31-item, 5-point, self-report paper-and-pencil instrument that measures patients' perception of the occurrence and distress of 14 symptoms: nausea, vomiting, pain, eating, sleep, fatigue, bowel elimination, breathing, coughing, concentration, lacrimation, changes in body temperature, appearance, and restlessness. Use of the instrument yields a total score for symptom experience, scores for symptom occurrence, scores for symptom distress, and subscale scores for six symptom categories: gastrointestinal, fatigue/restlessness, concentration, pain/discomfort, respiratory, and appearance. Reliability and validity were determined with well adults (n = 97), medical-surgical patients (n = 82), and oncology patients (n = 175). Findings revealed a Cronbach's alpha of 0.91 for symptom experience, 0.90 for symptom occurrence, and 0.76 for symptom distress. Cronbach's alpha for the subscales ranged from 0.38 for appearance symptoms to 0.83 for gastrointestinal symptoms. Inclusion of symptoms reported by patients with cancer strengthened content validity. A contrasted groups approach was used to demonstrate construct validity. PMID- 10673808 TI - Influencing nurses' knowledge, attitudes, and practice in cancer pain management. AB - The purpose of this study was to explore the effects of an education intervention on nurses' knowledge, attitudes, and practice in pain assessment and management over 3 months. The education intervention program was designed to change knowledge and influence the attitudes of registered nurses through a values clarification process using a conceptual framework based on a theory of reeducation. Participants in this descriptive, exploratory study were 53 nurses from six oncology units. Data were collected on their knowledge, attitudes, documentation practices, and analgesic choices in defined patient situations. The intervention was effective in changing the knowledge, attitudes, and behaviors of nurses in the study, but the effect was not maintained over time. Study findings suggest that further educational and organizational support is needed for effective practice in pain assessment and management. Further research should explore education programs that will maintain new knowledge over time. In addition, assessment of the effect that new knowledge has on the achievement of improved pain relief for patients should be explored in the future. PMID- 10673809 TI - Middle Eastern Asian Islamic women and breast self-examination. Needs assessment. AB - This exploratory, descriptive research study examined the knowledge and frequency of breast self-examination (BSE) among Middle Eastern Asian Islamic immigrant women residing in a major metropolitan U.S. city. The purposive sample consisted of 39 Middle Eastern Asian Islamic immigrant women ranging in age from 20 to 48 years (mean, 33; SD, 8.29). The sample was recruited from women who attended a local mosque. Data was collected by asking a list of seven questions based on Champion's BSE tool that assessed knowledge and frequency of BSE. Sociodemographic information also was collected. The results indicated that 33 women (85%) had heard of breast self-examination, and 29 women (74%) had not examined their breasts for lumps. None of the participants had examined her breasts monthly for lumps during the past year. Most of the women had not learned about BSE from a health professional, nor had they undergone a clinical breast examination (CBE). The results of this research show that Middle Eastern Asian immigrant women may be a population overlooked by health care professionals in the education of BSE. Suggestions to improve breast cancer screening practices among this population are provided. PMID- 10673810 TI - Meeting the information needs of adult daughters of women with early breast cancer. Patients and health care professionals as information providers. AB - This study explored the information needs of adult daughters whose mothers had early breast cancer and illustrated the information flows between the daughters and two information providers: patients and health care professionals (HCPs). Participants were 97 daughters, who completed a 30-time self-administered questionnaire, a tool designed to identify the information needs of daughters and their communication channels. Daughters also completed the Miller Behavioral Style Scale, a scale used to determine the information-seeking behavior of individuals under threat. Descriptive statistics revealed that the information flows between women with breast cancer and daughters participating in this study were good. Most of the daughters received information from their mothers and asked their mothers questions. Few daughters thought their mothers avoided giving them information. The vast majority of the daughters, however, sought information from sources other than their mothers, indicating that the mothers alone did not satisfy their information needs. More than half of the daughters participating in this study accompanied their mothers to their medical consultations and received information from HCPs. Logistic regression indicated that the communication flows between patients, HCPs, and adult daughters of women with breast cancer depended on a number of factors, in particular, on the amount of communication desired by women with breast cancer. PMID- 10673811 TI - Planning for implementation of a vendor-based clinical information system. Case study. PMID- 10673812 TI - The Internet: an effective tool for nursing research with women. AB - This article outlines the methodology of using the Internet to survey an international population of women about their perceptions of breast health education and screening. Issues to consider in planning and implementing the research project by Internet are presented. A large population of women from North America and elsewhere was reached through the establishment of a website with linkages to other sites frequented by women. Women who visited the website were asked to complete a questionnaire. Anonymity was guaranteed and simple instructions were provided at the site. Benefits, limitations, and tips for success in using the Internet as a research tool are presented. These investigators found the Internet to be an appropriate medium for health-related research that also garnered national and international media interest. The address for this website is http:@www.uwindsor.ca/breast.study/quest.htm. PMID- 10673813 TI - The effects of Internet-based distance learning in nursing. AB - Distance learning has been the turning point for accomplishing adult learning in nursing education. This article describes the development and structure of a distance-learning course used to deliver distance learning to the RN-BSN students at Yonsei University, College of Nursing. The distance-learning course was developed cooperatively by content experts, instructional designers, programmers, and graphic designers. The course content, "Growth and Development", was a computerized instructional course delivered using the Internet. The programming system was developed on the Web Server and Oracle DB through the Internet. The characteristics of adult learners--graduates with 3-year RN diplomas and working full-time--were considered during development of the course. For a semester, the students studied the growth and development of a person from infancy to adolescence through interactions with peers and instructors using alternative menus on the Internet. The course was evaluated from feedback of 60 RN-BSN students on their satisfaction with this distance-learning course in regard to instructional design, the arrangement and structure of instruction, and the function and feasibility of the courseware. When the self-reported questionnaire with 25 open questions was evaluated, students' general responses were relatively positive. Insufficient feedback from the professor, excessive time and difficulties experienced when connecting to the Internet, and the lack of information about related websites were primary negative responses. For an effective use of the distance-learning system, improvements to the telecommunications network service are crucial. School authorities should support the professors who are interested in developing distance-learning courses so that the courses can be developed with technical perspectives. More distance-learning courses applying interactive multimedia instructional design through the Internet should be developed with the improved network service in the future. PMID- 10673815 TI - Computer-based support groups. Nursing in cyberspace. AB - The focus of this article is on the nurse monitor role in a project whose overall goal is to use telecommunication technology to provide information and support to middle-aged rural women living with chronic illness. The impact of participation in these support groups on the women's psychosocial health is also discussed. The purpose of the project, the underlying conceptual framework of social support, a project overview, project philosophy and protocols, and the role of the Nurse Monitor are described. PMID- 10673814 TI - Integrating Web-based instruction into a graduate nursing program taught via videoconferencing. Challenges and solutions. AB - This article describes the methods used, problems encountered, and solutions that were generated while integrating Web-based instruction (WBI) into a Master of Science in Nursing Leadership and Case Management, and a Post-Master's Case Management certificate distance education program taught via video-conferencing. The writer describes the use of computer-mediated communication to enhance collaboration between and among students at 11 video-conferencing sites. The integration of WBI to support collaboration was successful because students and faculty could complete their academic activities in the locations and times that were most convenient to them. The tools that WBI provided were instrumental in helping students submit collaborative work that faculty often identified as far exceeding their expectations. PMID- 10673816 TI - The Internet: an underutilized tool in patient education. AB - Internet technology is helping to reshape patient education. An illustration of this is provided by data from a two-stage pilot study involving 100 senior citizens who received instruction on how to conduct health information searches on the Internet. The goals were to enable the seniors to assume an active role in their health care and to share their information with family and friends. In a Train-the-Trainer approach, 20 trainers received instruction on searching for health information on the Internet, and subsequently trained 100 senior citizen trainees. The study was conducted from October 1997 to June 1998. The average age of the senior trainees was 69. Most had a college education. The study results reveal a positive impact of the training on senior trainee confidence in using the computer and the Internet, conducting health information searches online, and sharing health care information with their physicians, families, and friends. Some gender and educational differences were noted. In a 90-day posttraining follow-up, 66% of the trainees continued to use the Internet, with 47% of them using it to search for health information. Two thirds of those who searched for health information on the Internet talked about it with their physicians, with more than half reporting they were more satisfied with their treatment as a result of their searches and subsequent discussion with their physicians. These findings are relevant to patient education in the nursing curricula of nursing students and nurse practitioners. Some suggestions are given to improve the effectiveness of the training program. PMID- 10673817 TI - Children of depressed mothers. PMID- 10673818 TI - Selective serotonin reuptake inhibitor-induced mydriasis. PMID- 10673819 TI - Olanzapine in Tourette's disorder. PMID- 10673820 TI - Olanzapine in Tourette's disorder. PMID- 10673821 TI - Secondary enuresis associated with obstructive sleep apnea. PMID- 10673822 TI - Sexual obsessions in obsessive-compulsive disorder. PMID- 10673823 TI - Language disorders: a 10-year research update review. AB - OBJECTIVE: To review the past 10 years of research in child language or communication disorders, which are highly prevalent in the general population and comorbid with childhood psychiatric disorders. METHOD: A literature search of 3 major databases was conducted. The child language literature, describing the domains of language development--phonology, grammar, semantics, and pragmatics- is reviewed. RESULTS: Disorders of grammar, semantics, and pragmatics, but not phonology, overlap significantly with childhood psychiatric disorders. Receptive language disorders have emerged as high-risk indicators, often undiagnosed. Language disorders and delays are psychiatric risk factors and have implications for evaluation, therapy, and research. However, they are often undiagnosed in child mental health and community settings. The research has focused mostly on monolingual English-speaking children. CONCLUSION: Awareness of basic child language development, delay, and deviance is crucial for the practicing child and adolescent psychiatrist, who must diagnose and refer relevant cases for treatment and remediation. Future research needs to address the growing language diversity of our clinical populations. PMID- 10673824 TI - Effectiveness of nonresidential specialty mental health services for children and adolescents in the "real world". AB - OBJECTIVE: Although many studies demonstrate the efficacy of a variety of treatments for child and adolescent psychiatric disorders, studies showing the effectiveness of such treatments in ordinary clinical settings have not been forthcoming. This report presents a study of the effectiveness of outpatient treatment in a community sample of 9- to 16-year-olds. METHOD: Four annual waves of data were collected from a representative sample of 1,422 children and their parents in the southeastern United States. Interviews were conducted with the Child and Adolescent Psychiatric Assessment to determine clinical status and the Child and Adolescent impact Assessment to measure the impact of psychiatric disorder on the lives of the children's families. RESULTS: Treated individuals were more severely disturbed and showed deterioration in their clinical status, even before they received treatment, indicating that comparisons with untreated individuals required controls not only for pretreatment clinical status, but for pretreatment clinical trajectory. A significant dose-response relationship was found between the number of specialty mental health treatment sessions received and improvement in symptoms at follow-up. However, no effect of treatment on secondary psychosocial impairment or parental impact was identified. CONCLUSIONS: Child and adolescent outpatient psychiatric treatment has positive effects on psychiatric symptoms, even when conducted outside the academic units where efficacy research usually takes place. The dose of treatment required to produce such effects (more than 8 sessions) suggests that attempts to limit child psychiatric treatment to very short-term interventions may be counterproductive. PMID- 10673825 TI - Dose effect in child psychotherapy: outcomes associated with negligible treatment. AB - OBJECTIVE: To compare the outcomes of children who received negligible amounts of outpatient treatment to children receiving more treatment. METHOD: A random regression longitudinal model was used to analyze outcomes of children (aged 5-17 years) from the Fort Bragg Evaluation Project. RESULTS: In examining several outcomes, the results show no statistically significant dose effect. CONCLUSIONS: Children receiving substantial amounts of treatment showed no better mental health outcomes than those receiving negligible amounts of treatment. The results do not support the existence of a dose effect consistent enough to guide clinicians, administrators, or policymakers. PMID- 10673826 TI - Commentary: more outcome studies are needed. PMID- 10673827 TI - Commentary: the dose effect in children's mental health services. PMID- 10673828 TI - Strengths of children and adolescents in residential settings: prevalence and associations with psychopathology and discharge placement. AB - OBJECTIVE: During the past few years there has been growing interest in developing strength-based approaches to services, particularly for children and adolescents. METHOD: This study assesses the prevalence of 30 strengths for a random sample of children and adolescents in residential placements in Florida. In addition, the relationship between strengths and clinical and functional characteristics is studied. RESULTS: Results suggest that there is substantial variation across individuals on the presence of strengths and the potential for development. Strengths were associated with symptoms, risk behaviors, and functioning. Level of strengths predicted success in the reduction of risk behaviors during the child/adolescent's stay. In addition, the level of strengths was independently associated with good dispositional outcomes. CONCLUSIONS: The findings provide further empirical support for the importance of strengths and the utility of an integrated model that considers both psychopathology and strengths in planning for children's services. PMID- 10673829 TI - National Institutes of Health Consensus Development Conference Statement: diagnosis and treatment of attention-deficit/hyperactivity disorder (ADHD) AB - Attention-deficit/hyperactivity disorder (ADHD) is a commonly diagnosed behavioral disorder of childhood that represents a costly major public health problem. Despite progress, ADHD and its treatment have remained controversial, especially the use of psychostimulants for both short- and long-term treatment. Although an independent diagnostic test for ADHD does not exist, there is evidence supporting the validity of the disorder. Studies (primarily short-term, approximately 3 months), including randomized clinical trials, have established the efficacy of stimulants and psychosocial treatments for alleviating the symptoms of ADHD and associated aggressiveness and have indicated that stimulants are more effective than psychosocial therapies in treating these symptoms. Because of the lack of consistent improvement beyond the core symptoms and the paucity of long-term studies (beyond 14 months), there is a need for longer-term studies with drugs and behavioral modalities and their combination. Although trials are under way, conclusive recommendations concerning treatment for the long term cannot be made at present. There are wide variations in the use of psychostimulants across communities and physicians, suggesting no consensus regarding which ADHD patients should be treated with psychostimulants, and thus the need for improved assessment, treatment, and follow-up. Furthermore, the lack of insurance coverage, preventing the appropriate diagnosis and treatment of ADHD, and the lack of integration with educational services are substantial barriers and represent considerable long-term costs for society. Finally, after years of clinical research and experience with ADHD, knowledge about the cause or causes of ADHD remain largely speculative. Consequently, there are no documented strategies for the prevention of ADHD. PMID- 10673830 TI - Commentary: the NIH ADHD Consensus Statement: win, lose, or draw? [ comment]. PMID- 10673831 TI - Treatment services for children with ADHD: a national perspective. AB - OBJECTIVE: To summarize knowledge on treatment services for children and adolescents with attention-deficit hyperactivity disorder (ADHD), trends in services from 1989 to 1996, types of services provided, service mix, and barriers to care. METHOD: A review of the literature and analyses from 2 national surveys of physician practices are presented. RESULTS: Major shifts have occurred in stimulant prescriptions since 1989, with prescriptions now comprising three fourths of all visits to physicians by children with ADHD. Between 1989 and 1996, related services, such as health counseling, for children with ADHD increased 10 fold, and diagnostic services increased 3-fold. Provision of psychotherapy, however, decreased from 40% of pediatric visits to only 25% in the same time frame. Follow-up care also decreased from more than 90% of visits to only 75%. Family practitioners were more likely than either pediatricians or psychiatrists to prescribe stimulants and less likely to use diagnostic services, provide mental health counseling, or recommend follow-up care. About 50% of children with identified ADHD seen in real-world practice settings receive care that corresponds to guidelines of the American Academy of Child and Adolescent Psychiatry. Physicians reported significant barriers to service provision for these children, including lack of pediatric specialists, insurance obstacles, and lengthy waiting lists. CONCLUSIONS: The trends in treatment services and physician variations in service delivery point to major gaps between the research base and clinical practice. Clinical variations may reflect training differences, unevenness in the availability of specialists and location of services, and changes in health care incentives. PMID- 10673832 TI - Predictors of panic attacks in adolescents. AB - OBJECTIVE: To identify risk factors for onset of panic attacks in adolescents, a prospective cohort design was used to evaluate the following risk factors: negative affectivity, female sex, anxiety sensitivity, and childhood separation anxiety disorder. These risk factors were also evaluated for predicting onset of major depression to test their specificity. METHOD: The sample consisted of 2,365 high school students assessed over a 4-year period. Assessments included self report questionnaires and structured clinical interviews. Cox proportional hazards models were used to evaluate risk. RESULTS: Consistent with previous studies, prior major depression predicted onset of panic attacks and a history of panic attacks predicted onset of major depression. After adjusting for the effects of prior major depression, negative affectivity and anxiety sensitivity, but not female sex or childhood separation anxiety disorder, predicted onset of 4 symptom panic attacks. However, female sex and negative affectivity but not anxiety sensitivity or childhood separation anxiety disorder predicted onset of major depression after adjustment for the effects of prior panic attacks. CONCLUSION: Negative affectivity appears to be a nonspecific risk factor for panic attacks and major depression, whereas anxiety sensitivity appears to be a specific factor that increases the risk for 4-symptom panic attacks in adolescents. PMID- 10673833 TI - Relationship between depression and substance use disorders in adolescent women during the transition to adulthood. AB - OBJECTIVES: To examine the continuity of substance use disorder (SUD) in adolescent women during the transition to adulthood and to assess psychosocial functioning associated with SUD. Furthermore, to examine concurrent and longitudinal relationships' between major depressive disorder (MDD) and SUD during this developmental transition. METHOD: One hundred fifty-five women, aged 17 to 19 years, were recruited from 3 high schools and were followed annually for 5 years. Comprehensive diagnostic and psychosocial assessments were performed with standardized instruments. The primary outcome measures included MDD and SUD during follow-up in those with and without a prior history of MDD or SUD, and psychosocial functioning associated with SUD. RESULTS: The 5-year incidence of SUD was 9.6% and, by the end of follow-up, 18.7% had a lifetime episode. Prior SUD significantly increased the risk for SUD diagnosis during the study. Co occurrence of MDD and SUD was high during adolescent and early adult years, with episodes of both disorders occurring in close temporal proximity. SUD also predicted MDD over time, but the reverse was not true. After controlling for the effects of MDD on social adjustment, SUD was associated with significant impairment in school functioning. CONCLUSIONS: These results suggest that the risk for new onset and recurrence of SUD is high during the developmental transition to adulthood. SUD during this developmental period is associated with significant school-related problems. The findings also suggest that SUD and MDD frequently co-occur during the post-high school transition in women. Given the significant psychosocial dysfunction associated with these illnesses, early detection of these problems and effective intervention are crucial. PMID- 10673834 TI - General and specific childhood risk factors for depression and drug disorders by early adulthood. AB - OBJECTIVE: To identify childhood risk factors that predict depression and drug disorders by early adulthood, distinguishing between general risk factors for both disorders and specific risk factors for each individual disorder. METHOD: Within a longitudinal community study (N = 360), familial and behavioral emotional characteristics were assessed in early childhood (ages 5, 6, and 9 years). At age 21, the Diagnostic Interview Schedule, version III-revised, provided lifetime diagnoses of major depression and drug abuse/dependence. RESULTS: Sibling substance use disorders predicted depression and drug disorders for both genders. Feelings of anxiety, depression, and peer rejection were general predictors for females. Specific risk factors for depression were parental depression and anxious/depressed behavior in both genders and peer problems for males. Specific risk factors for drug abuse/dependence were larger family size, lower socioeconomic status, hyperactivity, attention problems, and aggression. Parental substance abuse and having younger parents were specific risk factors for drug disorders in males. CONCLUSIONS: Familial and behavioral emotional risk factors for depression and drug disorders were primarily specific, suggesting separate pathways. The unique perspectives of multiple informants facilitate early identification. PMID- 10673835 TI - Competence and behavioral/emotional problems among Taiwanese adolescents as reported by parents and teachers. AB - OBJECTIVE: To investigate competence and behavioral/emotional problems among nonreferred adolescents in Taiwan, using a Chinese version of the Child Behavior Checklist (CBCL-C) and the Teacher's Report Form (TRF-C). The psychometric properties of these instruments and cross-cultural differences were also examined. METHOD: Parents of 854 junior high school students aged 12 to 16 years in Taipei, Taiwan, were asked to complete the CBCL-C. Among these students, 162 had their teachers' ratings of the TRF-C. RESULTS: The internal consistency and 1 month test-retest reliability were satisfactory for both the CBCL-C and TRF-C, which were moderately correlated. Both exploratory and confirmatory factor analysis provided some support for the validity of Achenbach's cross-informant model. Parents' reports showed that compared with their American counterparts, Taiwanese adolescents tended to have lower scores on most competence scales, higher scores on scales that reflect covert behavior problems, and lower scores on scales that reflect more overt behavior problems. However, teachers' reports showed no significant differences on most competence and behavior problem scales. CONCLUSION: The CBCL-C and TRF-C are useful tools for assessing the mental health status of Taiwanese adolescents. The cross-cultural differences in adolescent behavior problems are discussed. PMID- 10673836 TI - Effects of parent personality, upbringing, and marijuana use on the parent-child attachment relationship. AB - OBJECTIVE: To examine the predictors of the quality of the parent-child attachment relationship among a sample of 248 young adults with children. METHOD: In this longitudinal study, data were collected during early adulthood in 1992 and in 1996/1997 via a structured questionnaire. Using logistic regression and multiple regression analyses, the authors assessed the extent to which participants' personality attributes, substance use, and relationships with their mothers predicted the quality of the parent-child bond. RESULTS: Logistic regression models showed that participants with certain personality attributes (e.g., high sensitivity), less frequent marijuana use, or a close relationship with their mothers had a greater likelihood of having a close parent-child attachment relationship with their own children at a later time. Regression analysis also showed that the risk of earlier substance use on the parent-child relationship was offset by protective factors in the parents' personality domain. In addition, protective factors in the various parental domains synergistically interacted with a low frequency of marijuana use, relating to a closer parent child attachment relationship. CONCLUSIONS: The findings suggest that certain parenting styles are transmitted across generations and interventions in the personality and drug use domains can help increase the likelihood that parents will form close attachment relationships with their own children. PMID- 10673837 TI - Selective mutism and comorbidity with developmental disorder/delay, anxiety disorder, and elimination disorder. AB - OBJECTIVES: To assess the comorbidity of developmental disorder/delay in children with selective mutism (SM) and to assess other comorbid symptoms such as anxiety, enuresis, and encopresis. METHOD: Subjects with SM and their matched controls were evaluated by a comprehensive assessment of the child and by means of a parental structured diagnostic interview with focus on developmental history. Diagnoses were made according to DSM-IV. RESULTS: A total of 54 children with SM and 108 control children were evaluated. Of the children with SM, 68.5% met the criteria for a diagnosis reflecting developmental disorder/delay compared with 13.0% in the control group. The criteria for any anxiety diagnosis were met by 74.1% in the SM group and for an elimination disorder by 31.5% versus 7.4% and 9.3%, respectively, in the control group. In the SM group, 46.3% of the children met the criteria for both an anxiety diagnosis and a diagnosis reflecting developmental disorder/delay versus 0.9% in the controls. CONCLUSIONS: SM is associated with developmental disorder/delay nearly as frequently as with anxiety disorders. The mutism may conceal developmental problems in children with SM. Children with SM often meet diagnostic criteria for both a developmental and an anxiety disorder. PMID- 10673838 TI - Case study: possible traumatic stress disorder in an infant with cancer. AB - The posttraumatic stress disorder model has been used to describe some children's experience with cancer. This article presents the case of a 5-month-old infant in whom a neuroblastoma was diagnosed at age 2 weeks. His symptoms at the time of the psychiatric consultation were consistent with the criteria for traumatic stress disorder in infants from the ZERO TO THREE: Diagnostic Classification. This case study invites future clinical and research queries about traumatic stress disorder in infants with serious medical illnesses. PMID- 10673839 TI - Research questions and hypotheses. PMID- 10673840 TI - Genetics of childhood disorders: XI. Fragile X syndrome. PMID- 10673841 TI - Rickets and deprivation: a Nigerian study. AB - Under-fives in 461 households were assessed clinically to determine the prevalence of rickets in sub-urban and rural communities in the Sahel savanna. Overt rickets was found in 11 (2.4%) of households and abnormalities suggestive of rickets in 69 (14.9%). There were significant variations (p < 0.05) in the prevalence of rickets in association with ethnic grouping (higher in southerners and non-Kanuri, non-Hausa-Fulani northerners), religion (more prevalent among Christians), and mother's occupation and educational status (higher with working class mothers and mothers with at least a primary education). A significantly higher prevalence was also associated with late introduction (at more than seven months of age) of cereals to the infant's diet, more than one under-five in a household and presence of under-fives aged 13-43 months. In contrast, no significant variations in prevalence were observed in association with duration of breast feeding, use of multivitamins or cod liver oil, history of convulsions in under-fives, sex, nutritional status, or history of diarrhoea within a recall period of six months. Thus, rickets is common in under-fives in rural and sub urban communities in the Sahel savanna and may be related more to environmental and dietary factors than to culture and religion. Further studies are required to determine the relative roles of vitamin D or calcium deficiency to facilitate the planning and execution of a community-based intervention programme in the area. PMID- 10673842 TI - Obesity among Kuwait University students: an explorative study. AB - University students' dietary habits have been criticised for their nutritional inadequacy and faddism. Kuwait University students may face the risk of obesity because of affluence and modernization and the dynamic changes in their level of physical activity and caloric intake. This promoted a study of a random sample of 842 Kuwait University students for dietary and socioeconomic factors associated with obesity. Weight and height were measured to calculate the body mass index (BMI), which is the weight in kilograms divided by the height in meters squared (kg/m2). Obesity was classified into grade 1 and 2 (BMI > 25 and > 30 kg/m2). The associated factors studied and obtained through questionnaires included gender, age, marital status, parental obesity, education and occupation, dieting, last dental and health check-up, year of study, number of siblings (total, brothers and sisters), eating in between meals, high school and college GPA and major, exercising, number of regular meals eaten, obese relatives, those living at home, and servants, highest desired degree, birth order, having a chronic disease, countries prefer visiting, family income, governorate, and socioeconomic status (SES). Grade 1 and 2 obesity were found to be 32.0 and 8.9%, respectively. Factors that were found to be significantly associated with obesity included gender, age, marital status, obesity among parents, dieting, last physical check up, year of study, number of brothers, sisters and regular meals eaten and high school GPA. Logistic regression analysis revealed that the same factors significantly contributed to the development of obesity except the last four. The level of obesity among Kuwait University students is high. Obesity is a risk factor for a variety of chronic diseases. There is a need to address the challenge of instituting measures that would reduce the future ill-effects of obesity on young adults. It is widely believed that during the young adult years many important health habits are formed and set. It is at this stage (or earlier) that wellness and self-care programming for college students is essential and worthy of being explored and its efficacy assessed. PMID- 10673843 TI - Healthy housing: the role of the environmental health officer. AB - The relationship of health and housing has been well documented. There is less said about action that can be taken to deal with poor housing conditions. Environmental Health Officers in UK Government are key actors enforcing legislation relevant to housing conditions. Despite a century of legislative intervention in private sector housing conditions, the English House Condition Survey continues to report an excessive amount of poor conditions, and a particular decline in quality amongst the most disadvantaged in society who are increasingly accommodated in the private rented sector. This paper examines the role of poor housing in ill health and the difficulties faced in enforcing largely reactive legislative, arguing that wider changes are needed if the link between poor health and housing is to be broken. PMID- 10673844 TI - Is continued weight gain inevitable in type 2 diabetes mellitus? AB - Prevention and treatment of obesity are major clinical problems encountered in the management of Type 2 diabetes mellitus (DM); indeed, up to 90% of such patients are regarded as being overweight. Except for a brief period following diagnosis, when presumably enthusiasm to adopt lifestyle change is at its greatest, weight gain is generally progressive unless severe hyperglycaemia or complications intervene. Even a relatively modest weight loss of 10% can have major benefits in terms not only of reducing the risk of developing DM in the first place, but also in improving metabolic control after the disorder has become established. Behavioural therapy (BT) in combination with hypocaloric diet achieves weight loss in the short-term, but is poorly sustained in the long-term. Exercise has metabolic benefits beyond its rather minimal effects on short-term weight loss in that it may also aid long-term weight control. The difficulties encountered in maintaining lifestyle change do, however, suggest the need for ongoing intervention--perhaps including a regular period on a stricter dietary regimen (800-1000 kcalday-1), possibly a very low calorie diet (VLCD)(< 800 kcalday-1) or even the use of orlistat, a pancreatic lipase inhibitor which reduces the absorption of dietary fat. Realistically, the aim should be for long term weight stability. PMID- 10673845 TI - Possible health and safety problems in the use of novel plant essential oils and extracts in aromatherapy. AB - Aromatherapy is a branch of complementary or alternative therapy which is increasing in popularity, yet has scant scientific credibility. Aromatherapy should be defined as treatment using odours and practised as such. However, essential oils are usually used in conjunction with massage and often combined with counselling of some kind. The use of most commonly-used essential oils in massage is seldom dangerous, as they have low systemic toxicity, especially when used at 2% dilution (provided they are not adulterated); however their safety during pregnancy, childbirth and babies has not been clearly demonstrated. Sensitisation, however, is a growing concern. Some aromatherapists are now introducing novel plant essential oils, extracts and phytols into their massage routine many of which have no odour and are potentially toxic. The possible dangers of these plant products are therefore discussed using specific example. PMID- 10673846 TI - Cognitive behaviour therapy: its evolution and basic principles. AB - The historical and theoretical background to Cognitive Behaviour Therapy (CBT) is outlined. The main therapeutic approaches of Behaviour Therapy (BT) and Cognitive Therapy (CT) are described--and an overview is provided of the applications of these approaches to clinical problems. The relationships between BT, CT and CBT are clarified. PMID- 10673848 TI - Overweight and obesity in the Arabian Peninsula: an overview. AB - Obesity greatly increases the risks of developing diseases, including diabetes mellitus, hypertension, dyslipidaemia, coronary artery disease, and some cancers. At least one-third of Arabs are obese, as defined by body mass index (BMI) more than 30 kg/m2 and this figure is rising steadily despite increased interest in fitness. Women have particularly high rates of obesity. Obesity is clearly associated with increased risk for mortality, but there has been controversy regarding optimal weight with respect to mortality risk. A review of recent studies on the prevalence of obesity among population of the Arabian Peninsula and the evaluation of the health risk of obesity is presented in this paper. The prevalence of obesity ranges between 16-25% in men and 17-43% in women. The most prevalent chronic diseases related to obesity in these populations are diabetes, hypertension and coronary heart disease (CHD). PMID- 10673849 TI - Occupant injury protection in automobile collisions. AB - Modern technology has produced automotive vehicles that have become both a luxury and a necessity in modern civilization. They have become highly useful, even more varied in form and function, and capable of high speeds on crowded roadways. One unfortunate consequence is the high frequency of accidents and the greater severity of injuries when collisions do occur. In response, modern technology has produced a variety of safety and health features, devices and designs intended for better occupant protection on in high speed vehicles. Injury reduction has become a prime design objective, but there are residual risks, which, as technology evolves, require effective communication to those risk. There can be little risk avoidance behavior without awareness of the hazards and effective communication to the vehicle occupant, as to what could and should be done for self-protection. For example, one out of three drivers apparently fails to understand the function of head restraints, few understand the 'safe zone' posture required for air bags and many believe safety features should be adjusted only for comfort. Some of the current residual injury producing problems in occupant systems are specifically described here in order to illustrate what is needed in terms of both design remedies and health promotion activities. PMID- 10673847 TI - Hospital transformation and organisational learning. AB - Kwong Wah Hospital was founded by the charity organisation Tung Wah Group of Hospitals some 88 years ago, with management transfer to the Hong Kong Hospital Authority in 1991. Capitalizing both from the traditional caring culture of its founder, as well as opportunities in the new management environment, the hospital has scored remarkable successes in service quality, community partnership, organisational effectiveness, and staff development. Underpinning these transformations were Structure, Process, People, and Culture strategies. The learning imperative is heavily mandated or the success of each of these strands of development. Indeed, the embodiment of a learning organisation culture provides the impetus in sustaining the change momentum, towards achieving the Vision of becoming a 'Most Preferred Hospital' in Hong Kong. PMID- 10673850 TI - The 1998 APHA Annual Lecture. What the UK can teach the US about health care. PMID- 10673851 TI - Lessons to be learned: a case study approach: metastatic bronchogenic carcinoma presenting as a gluteal abscess. AB - It is well known that bronchogenic carcinoma frequently metastasises to the bony skeleton, but it is most unusual for it to present in the form of a musculoskeletal abscess. Presented here is the case report of a patient with what appeared initially to be a right sided gluteal abscess and which turned out to be the metastasis from a bronchogenic carcinoma. The Magnetic Resonance Image (MRI) scan carried out proved to be very helpful in arriving at a probable clinical diagnosis; however, it was histopathological studies of the abscess wall itself that ultimately gave the definitive diagnosis. We believe that this may represent one of the first documented cases in which on MRI scan has been used to confirm the presence of a gluteal abscess. PMID- 10673852 TI - [Current diagnostic method, prognosis estimation and therapy of papillary thyroid cancer: recommendations of the medical universities and the National Oncologic Institute of Budapest]. AB - Physical examination, cervical ultrasonography (US) and aspiration cytology are the mainstays of the preoperative diagnostics of papillary thyroid carcinoma. For the staging of suspected malignant cases, cervical and mediastinal CT (MRI for inconclusive results) is indicated before any surgery. The end-result of primary treatment is assessed by total-body iodine scintigraphy and the serum human thyroglobulin (hTG) level. For long-term follow-up, physical examination and the serum hTG level are the most reliable tools (6-monthly), supplemented by cervical US and chest X-ray (yearly), and total-body iodine scintigraphy (2-yearly). If these furnish positive results, further examinations may be indicated. In suspected relapses of hTG non-producing and iodine non-accumulating papillary carcinomas, 201thallium chloride or 99mTc-sesta-MIBI (methoxy-isobutyl isonitrile) scintigraphy, and positron emission tomography with 18fluoro deoxyglucose or 11C-methionine may be of help. For estimation of the prognosis (cause-specific survival) of the patients, the MACIS score system of the Mayo Clinic is widely accepted, the patients being divided into low-risk and intermediate/high-risk categories. The recommended standard surgical intervention is near-total thyroidectomy (2-4 g residual glandular tissue left at the upper pole of the less-involved lobe), with a central cervical lymph node dissection for diagnostic purposes. In cases of lymph node dissemination, dissection (radical, modified radical, selective or microdissection) of any of the involved compartments (central, right or left cervical, or upper mediastinal) is indicated for therapeutic reasons, the method of which is depending on the extent of the metastatic involvement. Following adequate surgical intervention, no adjuvant radioiodine therapy is indicated for low-risk cases with a tumour of less than 1 cm diameter. For other low-risk or intermediate/high-risk patients, radioiodine ablation (R0N0M0) or a therapeutic radioiodine dosage (R2N1M1) is indicated. In cases at high-risk of local/regional relapse and in radioiodine non-accumulating tumorous cases, external radiotherapy may be applied. Thyroid hormone medication in a TSH suppressive dose is indicated during the first 5 postsurgical years: the goal is to achieve a TSH level below 0.1 (determined by a 3rd generation assay). If no relapse occurs or the case is a low-risk one, following the 5 years, it is enough to maintain the TSH level in a subnormal range (0.1-0.3). PMID- 10673853 TI - [Prevalence of mood and anxiety disorders in the Hungarian adult population]. AB - The prevalence rates of affective and anxiety disorders in the Hungarian adult population were assessed with a well-structured questionnaire which has been successfully applied in several multinational epidemiological studies. The Hungary material showed significantly higher lifetime and period prevalence rates of bipolar disorders than is found in most of related literature. However, the frequency of other affective disorders and the anxiety states strongly corresponded with international findings. PMID- 10673854 TI - [Effect of fibrates on lipid metabolism and the blood coagulation system]. AB - The authors summarised the complex effects of fibrates on atherogen dyslipidaemia and the blood coagulation system, as well as the results of previous intervention studies with fibrates. They also give an overview of the favourable molecular effects of fibrates and the role of peroxysoma proliferator activated receptor alpha (PPAR-alpha). PMID- 10673855 TI - [Competitive polymerase chain reaction: a new method for measuring N-myc amplification]. AB - The highly variable biological behaviour of neuroblastoma, a neoplasm which belongs to the family of primitive neuroectodermal tumours, is determined by its molecular pathological characteristics. Among them amplification of the N-myc gene is the most important factor in both therapeutic and prognostic points of view. Value of the amplification can be determined by different methods. The latest of them is the competitive polymerase chain reaction (PCR), essence of which is the parallel reaction of the target N-myc gene (exon 3.) and the endogen competitor CF gene (exon 3.) in the same reaction solution. The authors applied the method on 11 neuroblastoma cases diagnosed between 1994 and january of 1999. In three cases the amplification was determined also by fluorescens in situ hybridization (FISH). Six of the 11 cases were detected to have more than 10 fold, two of the six about 100-fold N-myc amplification. Results of the PCR and FISH correlated well. The two methods applied by the authors complete each other and are appropriate for determining the gene amplification which gives valuable prognostic and therapeutic information about the examined tumour. PMID- 10673856 TI - [Refsum disease]. AB - For the first time in literature the authors interpret the pathography of Refsum's disease, in the case of their patient, as pseudo-hypervitaminosis A. The biochemical basis of the clinical picture is a defect in the activity of phytanic acid-alpha-hydrolase belonging to the peroxisomal system. As a consequence, phytanic acid accumulates in the serum and in the parenchymal tissues. Retinol, an alcohol with high molecular weight, is a natural ligand of nuclear RXR (retinoid-X-receptor), which plays an important role in the regulation of peroxisoma synthesis. In Refsum's disease the phytanic acid accumulated because of the enzyme defect competes with the biotransformation derivates (all-trans retinoic acid, 9-cis-retinoic acid) of the all-trans-retinol (vitamin A) for the nuclear RX receptor binding sites, and as a very potent receptoractivator it causes the intestinal symptoms of hypervitaminosis A. The authors review the procedure of fatty-acid chromatography necessary for the establishment of the diagnosis and discuss--in addition to dietary restrictions--recent therapeutic possibilities, like plasmapheresis, cascade filtration, lipapheresis and oral batylalcohol treatment. PMID- 10673857 TI - Information management and technology (IM&T): can the clinicians get there before government? PMID- 10673858 TI - Problems associated with manpower. PMID- 10673859 TI - An important anniversary. PMID- 10673860 TI - Surgical training for overseas doctors: an historical review and proposals for the future. PMID- 10673861 TI - Surgical training after MRCS: the SHO III gap. PMID- 10673862 TI - A recurring theme. PMID- 10673863 TI - Basic surgical skills courses: an educational success story. PMID- 10673864 TI - The implications of cancer therapy for oral health: a report of a meeting held on 12 March 1999. PMID- 10673865 TI - Where does research fit in? PMID- 10673866 TI - Teaching and research training should be integrated with clinical training. PMID- 10673867 TI - Cost effectiveness of general anaesthesia: inhalation vs i.v. PMID- 10673868 TI - Less is more ... using systolic pressure variation to assess hypovolaemia. PMID- 10673869 TI - Ventilatory frequency variability in spontaneously breathing anaesthetized subjects. AB - During spontaneous breathing general anaesthesia, inspiration is generally started by a signal related to preceding cardiovascular activity. This phenomenon, 'cardioventilatory coupling', contributes to the variation in ventilatory frequency. However, the detailed, breath-to-breath timing relationship between heart beat and inspiratory onset is complex, with at least four distinct patterns (designated patterns I-IV). These coupling patterns are defined according to the particular breath-to-breath change in: (a) entrainment ratio and (b) coupling interval, the interval between inspiratory onset and the preceding initiating heart beat. We have examined the relationship between coupling and timing of breathing in adult subjects breathing spontaneously during general anaesthesia. The heart rate-ventilatory frequency interaction was explored by identifying the distribution of different coupling patterns in a plot of heart rate vs ventilatory frequency (the HR/f plot) and analysing the variation in breathing frequency during each coupling pattern by differentiating between changes in entrainment ratio from changes in coupling interval. We observed that: (i) coupling patterns are distributed within specific regions of the HR/f plot; (ii) specific patterns of variation in breathing are associated with each coupling pattern; (iii) this variation is a consequence of the balance between changes in entrainment ratio and coupling interval; (iv) coupling was invariably present at low breathing frequencies; and (v) the inverse non-linear relationship between ventilatory frequency and variation is largely a consequence of changing coupling pattern with ventilatory frequency. Coupling explains much of the breath-to-breath variability of ventilatory frequency during anaesthesia, and may be relevant to the phenomena of hypoventilation, central apnoea and ventilatory arrhythmia. A hypothesis concerning the generation of coupling patterns is presented. PMID- 10673870 TI - A multicentre comparison of the costs of anaesthesia with sevoflurane or propofol. AB - Day-case anaesthesia requires rapidly eliminated anaesthetics which are relatively expensive. This multinational, multicentre European study assessed the relative costs of propofol or sevoflurane anaesthesia in 211 patients. Anaesthesia was induced and maintained with propofol in group 1, with propofol and sevoflurane in group 2, and with sevoflurane in group 3. Drug and delivery costs were calculated in US$. Induction of anaesthesia was fastest in groups 1 and 2, although spontaneous ventilation resumed earliest in group 3. Emergence times and times at which patients were fit for discharge were similar in all groups. Group 2 had the lowest costs based on actual drug use (mean $14.2 (SEM 0.8) vs $18.7 (0.8) and $17.3 (0.8) in groups 1 and 3, respectively). Anaesthetic drug wastage and disposable costs were highest in group 1 and lowest in group 3. Consequently, total costs were highest in group 1 ($31.9 (0.9)) compared with groups 2 ($19.7 (0.9)) and 3 ($18.8 (0.9)). Although we observed increased nausea and vomiting in groups 2 and 3 and reduced patient satisfaction in group 3, these differences should be balanced against the greater cost of propofol anaesthesia. PMID- 10673871 TI - Compound A does not accumulate during closed circuit sevoflurane anaesthesia with the Physioflex. AB - We have investigated inspiratory and end-tidal gas composition during sevoflurane anaesthesia in a closed circle system with continuous gas flow (70 litre min-1, Physioflex) to determine possible accumulation of sevoflurane degradation products. During five abdominal operations in adults lasting more than 2 h, anaesthesia was maintained with an end-tidal concentration of 2% sevoflurane in 40% oxygen-air. The circle included an absorbing canister filled with 1 litre of fresh soda lime. Samples were obtained at the end of an expiration from the tracheal tube and from the inspiratory limb before, and at selected times after, addition of sevoflurane. The temperature of soda lime was 24.7 +/- 0.7 degrees C at the beginning and reached a maximum of 31.2 +/- 1.0 degrees C after 20-30 min, followed by a plateau. Inspiratory compound A (CH2F-O-C(= CF2)(CF3)) 3-8 ppm was detected after 10 min, but did not accumulate in the circle over 2 h without flushing. Expired concentrations were consistently lower with 1.5-3 ppm signalling absorption by patients. Calculated total amounts absorbed over 2 h varied between 2.0 and 7.2 ppm h. Other degradation products such as compound B or methanol were not detected. In summary, we did not detect sevoflurane metabolites with soda lime in significant amounts during closed circle anaesthesia with the Physioflex. The observed concentrations of compound A were below the threshold of nephrotoxicity in rats by a factor of more than 20. PMID- 10673872 TI - Comparison of sevoflurane-nitrous oxide and propofol-alfentanil-nitrous oxide anaesthesia for minor gynaecological surgery. AB - We studied 44 patients undergoing minor gynaecological surgery, anaesthetized in random order with sevoflurane-nitrous oxide or propofol-alfentanil-nitrous oxide. Operating conditions, recovery and postoperative nausea and vomiting (PONV) were assessed. For postoperative analgesia, all patients were given ketoprofen 100 mg rectally at the end of anaesthesia. Patients and gynaecologists were equally satisfied with both anaesthetic techniques. Patients given propofol woke up (3.5 vs 6.5 min), became orientated (5.0 vs 7.5 min) and were able to walk (57 vs 69 min) significantly (P < 0.05) earlier than those given sevoflurane, but there were no differences in times to achieve home readiness (166 vs 149 min) or in psychomotor recovery between the two groups. Intrauterine bleeding and PONV were more common with sevoflurane (incidence of PONV 64%) than with propofol anaesthesia (incidence of PONV 5%). We conclude that propofol-alfentanil is preferable to sevoflurane in ultra-short anaesthesia for minor gynaecological surgery. PMID- 10673873 TI - Burst suppression or isoelectric encephalogram for cerebral protection: evidence from metabolic suppression studies. AB - Metabolic suppression may have a role in cerebral protection. It is often assumed that the cerebral metabolic and protective effects of qualitative burst suppression are similar to those of the isoelectric encephalogram (EEG). We have examined the effect of different degrees of EEG suppression on blood flow and oxygen difference during general anaesthesia. We studied 11 patients undergoing general anaesthesia for resection of acoustic neuromas. The study was performed after surgery with propofol and remifentanil anaesthesia. Transcranial Doppler ultrasonography and jugular bulb venous saturations were measured at values of EEG suppression: 0%, 50% and 100% (isoelectric EEG). Data from nine patients were suitable for analysis. There were no significant differences in mean arterial pressure, heart rate or PaCO2 during EEG activity, 50% burst suppression ratio or isoelectric EEG. There was a significant decrease in middle cerebral artery flow velocity (vmca) with increasing EEG suppression (0% suppression, mean 38 (SEM 4) cm s-1; 50% suppression, 29 (3) cm s-1; and 100% suppression, 24 (2) cm s-1; P < 0.05). Jugular bulb venous saturations did not change consistently with the change in EEG activity, indicating intact flow-metabolism coupling. We conclude that the degree of EEG suppression had a significant effect on blood flow. If flow-metabolism coupling is maintained, the assumption that cerebral metabolism during 50% EEG burst suppression is equivalent to isoelectric EEG may not be justified. If cerebral protection is related to brain metabolism, then an isoelectric EEG may give more cerebral protection than 50% burst suppression. PMID- 10673874 TI - Benzodiazepines and postoperative cognitive dysfunction in the elderly. ISPOCD Group. International Study of Postoperative Cognitive Dysfunction. AB - Postoperative cognitive dysfunction (POCD) has been attributed to long-acting sedatives. We hypothesized that diazepam and its active metabolites could be detected in blood after surgery and correlated with POCD, 1 week after surgery in elderly patients. We studied 35 patients, 60 yr or older, undergoing abdominal surgery with general anaesthesia, including diazepam. Neuropsychological tests were performed before surgery and at discharge, where blood concentrations (free fraction) of benzodiazepines were also measured. POCD was found in 17 patients (48.6%). Diazepam or desmethyldiazepam was detected in 34 patients; median postoperative blood concentrations were 0.06 and 0.10 mumol kg-1, respectively. In a multiple regression analysis considering age, duration of anaesthesia and blood concentrations of diazepam and desmethyldiazepam, only age was found to correlate with the composite z-score (F test, P < 0.01). The postoperative cognitive dysfunction we found in elderly patients after operation could not be explained by benzodiazepine concentrations detected in blood. PMID- 10673875 TI - Midazolam premedication and thiopental induction of anaesthesia: interactions at multiple end-points. AB - We have studied the effects of midazolam premedication on multiple anaesthetic end-points (hypnotic, loss of verbal contact (LVC); motor, dropping an infusion flex or bag (DF); analgesic, loss of reaction to painful stimulation (LRP); and EEG, attainment of burst suppression (BUR)) during induction by slow thiopental infusion at a rate of 55 mg kg-1 h-1. Patients received midazolam 0.05 mg kg-1 i.v. (group TM, n = 12) or no midazolam (group T0, n = 13). ED50 and ED95 values and group medians for times and doses at the end-points were measured. Midazolam premedication reduced significantly thiopental ED50 and ED95 values at all end points (exception for ED95 for BUR). Potentiation was greatest for the motor end point (dropping the infusion bag (DF)) (ED95 +52%, ED50 +23%, median +39%), and smallest for painful stimulation (LRP) (median +18%; ED50 +13%). For LRP and DF, premedication was associated with significant, non-parallel increases in the slope of the thiopental dose-response curves, resulting in marked potency ratio changes from ED50 to ED95 (LRP +31%, DF +29%). There were no such increases for LVC or BUR. The interaction between midazolam and thiopental varied with the anaesthetic end-point and may also depend on the dose of thiopental. Our data suggest that the mechanism of interaction between midazolam premedication and thiopental was different for motor effects or analgesia (DF, LRP) compared with hypnotic effects or cortical depression (LVC, BUR), in agreement with the different central nervous system substrates underlying these distinct anaesthetic end-points. PMID- 10673876 TI - Use of tranexamic acid for an effective blood conservation strategy after total knee arthroplasty. AB - We have investigated the effect of treatment with tranexamic acid, an inhibitor of fibrinolysis, on blood loss, blood transfusion requirements and blood coagulation in a randomized, double-blind, placebo-controlled study of 42 patients after total knee arthroplasty. Tranexamic acid 15 mg kg-1 (n = 21) or an equivalent volume of normal saline (n = 21) was given 30 min before surgery and subsequently every 8 h for 3 days. Coagulation and fibrinolysis values, blood loss and blood units administered were measured before administration of tranexamic acid, 8 h after the end of surgery and at 24 and 72 h after operation. Coagulation profile was examined (bleeding time, platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT), plasminogen, beta thromboglobulin and fibrinogen). Fibrinolysis was evaluated by measurement of concentrations of D-dimer and fibrinogen degradation products (FDP). Total blood loss in the tranexamic acid group was 678 (SD 352) ml compared with 1419 (607) ml in the control group (P < 0.001), and occurred primarily during the first 24 h after surgery. Thirteen patients received 1-5 u. of packed red blood cells in the control group compared with two patients in the tranexamic acid group, who received 3 u. (P < 0.001). Postoperative packed cell volume values were higher in the tranexamic acid group despite fewer blood transfusions. Postoperative concentrations of plasminogen were decreased significantly in the tranexamic acid group (P < 0.001). Platelet count, PT, aPTT, bleeding time, beta-thromboglobulin, fibrinogen and FDP concentrations did not differ between groups, but D-dimer concentrations were increased in the control group. Thromboembolic complications occurred in two patients in the control group compared with none in the tranexamic acid group. PMID- 10673877 TI - Effect of cardiopulmonary bypass on serum procalcitonin and C-reactive protein concentrations. AB - We have measured serum procalcitonin (PCT) concentrations after cardiac surgery in 36 patients allocated to one of three groups: group 1, coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) (n = 12); group 2, CABG without CPB (n = 12); and group 3, valvular surgery with CPB (n = 12). Serum PCT and C reactive protein (CRP) concentrations were measured before operation, at the end of surgery and daily until postoperative day 8. Serum PCT concentrations increased, irrespective of the type of cardiac surgery, with maximum concentrations on day 1: mean 1.3 (SD 1.8), 1.1 (1.2) and 1.4 (1.2) ng ml-1 in groups 1, 2 and 3, respectively (ns). Serum PCT concentrations remained less than 5 ng ml-1 in all patients. Concentrations returned to normal by day 5 in all groups. To determine the effect of the systemic inflammatory response (SIRS) on serum PCT concentrations, patients were divided post hoc, without considering the type of cardiac surgery, into patients with SIRS (n = 19) and those without SIRS (n = 17). The increase in serum PCT was significantly greater in SIRS (peak PCT 1.79 (1.64) ng ml-1 vs 0.34 (0.32) ng ml-1 in patients without SIRS) (P = 0.005). Samples for PCT and CRP measurements were obtained from 10 other patients with postoperative complications (circulatory failure n = 7; active endocarditis n = 2; septic shock n = 1). In these patients, serum PCT concentrations ranged from 6.2 to 230 ng ml-1. Serum CRP concentrations increased in all patients, with no differences between groups. The postoperative increase in CRP lasted longer than that of PCT. We conclude that SIRS induced by cardiac surgery, with and without CPB, influenced serum PCT concentrations with a moderate and transient postoperative peak on the first day after operation. A postoperative serum PCT concentration of more than 5 ng ml-1 is highly suggestive of a postoperative complication. PMID- 10673878 TI - Effect of tropisetron on vomiting during patient-controlled analgesia in children. AB - Patient-controlled analgesia (PCA) is associated with a high incidence of vomiting which is distressing and interferes with postoperative recovery. Tropisetron, a long-acting selective 5-HT3 receptor antagonist, has been shown to be effective in preventing nausea and vomiting associated with PCA use in adults and chemotherapy in children. We assessed the efficacy of prophylactic intraoperative administration of tropisetron on the incidence of vomiting in children using morphine PCA. We studied 58 patients, allocated randomly to receive tropisetron 0.1 mg kg-1 to a maximum of 5 mg, or normal saline. Children who received tropisetron had an incidence of vomiting during the first 24 h after operation of 22% compared with 66% in the control group (P = 0.001). In addition, the severity of vomiting was less in the tropisetron group with only one child (4%) vomiting more than twice compared with nine (31%) in the control group (P = 0.01). We conclude that tropisetron is efficacious in reducing the incidence and severity of postoperative vomiting in children using PCA. PMID- 10673879 TI - Comparison of ondansetron and cyclizine for prevention of nausea and vomiting after day-case gynaecological laparoscopy. AB - We have compared ondansetron 4 mg i.v. and cyclizine 50 mg i.v., in a double blind, randomized, placebo-controlled study for the prevention of postoperative nausea and vomiting (PONV) for 24 h after day-case gynaecological laparoscopy. Compared with placebo (n = 58), ondansetron (n = 60) and cyclizine (n = 57) reduced significantly the incidence of moderate or severe nausea (30% and 23% vs 52%; P = 0.02 and P = 0.001, respectively) and requirement for escape antiemetic (28% and 16% vs 47%; P = 0.04 and P < 0.001, respectively) before discharge from hospital. There were no significant differences in PONV after discharge. Significantly more patients suffered no PONV before and after discharge after ondansetron and cyclizine compared with placebo (31% and 33% vs 12%; P = 0.02 and P < 0.01, respectively). For diagnostic laparoscopy (n = 74), fewer patients received escape antiemetic after cyclizine than after ondansetron (4% vs 37%; P < 0.01); for laparoscopic sterilization (n = 101), both antiemetics were equally effective. Ondansetron and cyclizine both reduced severe and moderate nausea and the need for antiemetic therapy after day-case gynaecological laparoscopy. PMID- 10673880 TI - The intubating laryngeal mask airway compared with direct laryngoscopy. AB - We have compared the ability of naive intubators to intubate the trachea using a laryngoscope and an intubating laryngeal mask airway (ILMA) after receiving basic training, in a randomized, prospective, crossover study in 60 patients. Ventilation of the lungs via the ILMA was also compared with ventilation with a face mask. There was no significant difference in successful intubation between the techniques (38 of 89 with the ILMA and 33 of 93 with direct laryngoscopy; ns). In patients in whom participants failed in their intubation attempts with the ILMA, investigators achieved success in 89% (16 of 18). Satisfactory ventilation was more common with the ILMA (50 of 51) than with the face mask (43 of 60) (P = 0.0001). A total of 98% (89 of 91) of ILMA were inserted successfully, with a mean insertion time of 19.6 s, and 78% (69/89) of these insertions were achieved in less than 26 s. The ILMA may be useful for emergency oxygenation and ventilation, but these results do not support its use for intubation by those not trained in advanced airway management and ILMA use. PMID- 10673881 TI - Arterial to inspired partial pressure ratio of halothane, isoflurane, sevoflurane and desflurane in rats. AB - The inspired partial pressure of an anaesthetic is often used as an index of arterial partial pressure in small animal experiments. We have investigated the influence of anaesthetic solubility on the ratio of arterial to inspired partial pressure in 24 rats, allocated randomly to receive halothane, isoflurane or desflurane at four different inspired concentrations. The arterial partial pressure of the volatile agent was measured by two-stage headspace analysis using a gas chromatograph calibrated with the same gas used to calibrate the Datex Capnomac that measured the inspired concentration. Mean values of arterial to inspired ratio at the lowest concentrations were 0.60 (95% confidence intervals 0.50, 0.71) for 0.8% halothane, 0.54 (0.38, 0.69) for 0.8% isoflurane, 0.72 (0.59, 0.86) for 1.5% sevoflurane and 0.71 (0.54, 0.87) for 4% desflurane. Analysis of variance showed a significant effect of anaesthetic agent (P = 0.008) on the arterial to inspired ratio. Thus volatile anaesthetic agents do not demonstrate a fixed arterial to inspired ratio in rats. PMID- 10673882 TI - Effects of propofol on vascular reactivity in isolated aortae from normotensive and spontaneously hypertensive rats. AB - We have investigated the effects of propofol 50 mumol litre-1 on contractile and relaxant responses in experimental hypertension and assessed endothelial modulation of these responses. Propofol attenuated norepinephrine-induced contraction of endothelium-intact and endothelium-denuded rings from both Wistar Tokyo (WKY) and spontaneously hypertensive rats (SHR). The effect was significantly greater in endothelium-intact aortae from SHR than in those from WKY rats. Propofol markedly attenuated AVP-induced contraction in aortae from both WKY and SHR. Propofol attenuation of norepinephrine contraction was also observed in rings from both SHR and WKY rats incubated with L-NAME. Propofol attenuation of norepinephrine contraction was suppressed by indomethacin in aortae from SHR but not in those from WKY rats. These results suggest that: (1) propofol attenuated vascular contraction of isolated aortae from SHR in part by a mechanism dependent on events distal to the receptor site (norepinephrine, arginine vasopressin); (2) the effect of propofol on contraction in SHR, observed in the presence of nitric oxide synthase inhibitors but not cyclooxygenase inhibitors, was consistent with either propofol induction of vasodilating cyclooxygenase metabolites from the endothelium or propofol inhibition of vasoconstricting cyclooxygenase metabolites. PMID- 10673883 TI - Measurements of systolic time intervals using a transoesophageal pulsed echo Doppler. AB - Measurement of systolic time intervals (STI), an index of left ventricular (LV) systolic function, is usually labour intensive and requires considerable expertise to perform accurately. We have evaluated the accuracy of an automated, continuous and non-invasive STI measurement technique using a descending aortic blood velocity Doppler signal obtained using a transoesophageal echo-Doppler system (TEDS) and an ECG signal. STI were measured in adult pigs using a transoesophageal probe (4 x 4 mm pulsed wave Doppler transducer, 5-MHz frequency and a 3 x 3 mm echo transducer, 10-MHz frequency) associated with an ECG recorder. Measurements were performed at baseline and after injection of esmolol and dobutamine. TEDS data were compared with those obtained by one-line recordings of the electrocardiogram and the central aortic arterial pressure wave. Similar mean values were observed for pre-ejection period (PEPI), LV ejection time (LVET) and PEP/LVET with the two methods. Agreement between the methods (Bland and Altman's test) was excellent with 95% confidence intervals for PEP, LVET and PEP/LVET of -7.17 to +1.37 ms, -12.64 to +0.24 ms and -0.033 to +0.028, respectively. We conclude that the combination of descending aorta blood velocity Doppler and ECG signal is an alternative technique for non-invasive and objective measurement of STI, allowing continuous monitoring of LV systolic function. PMID- 10673884 TI - Adverse effects of cannabis and cannabinoids. PMID- 10673885 TI - Comparison of breathing methods for inhalation induction of anaesthesia. AB - We studied healthy female patients, allocated randomly to three breathing regimens for induction of anaesthesia using sevoflurane and oxygen from a co axial Mapleson D breathing system and a mask, to test the hypothesis that rebreathing reduces the incidence of apnoea associated with loss of consciousness. The non-rebreathing group received sevoflurane in oxygen 6 litre min-1 from the start, doubling in concentration from 0.5% to 8% every 3 breaths. The second group received oxygen 6 litre min-1 for 1 min before sevoflurane was introduced, and the third group received oxygen 3 litre min-1 for 1 min before sevoflurane. The incidence and duration of apnoea were assessed using pneumotachograph and impedance pneumograph recordings, and time to induction of anaesthesia (weight drop) was measured from the time the breathing sequence was started. There was no difference in these times, which were mean 121 (95% confidence values 91-160) s, 117 (69-201) s and 125 (76-192) s, respectively. There was a significant difference in the incidence of apnoea. No apnoea occurred during induction using oxygen 3 litre min-1. Apnoea occurred in five of 15 patients who did not receive oxygen before sevoflurane and in four of 13 who received oxygen 6 litre min-1 (P < 0.05). No patient showed a reduction in pulse oximeter saturation. We conclude that inhalation induction of anaesthesia can be performed reliably in approximately 3 min using sevoflurane in oxygen 3 litre min 1. PMID- 10673886 TI - Effect of a remifentanil bolus dose on the cardiovascular response to emergence from anaesthesia and tracheal extubation. AB - We have examined the effect of remifentanil on the haemodynamic response to emergence from anaesthesia and tracheal extubation in 40 ASA I-II female patients undergoing diagnostic laparoscopy, in a randomized, double-blind study. All patients received a standard general anaesthetic comprising propofol, vecuronium and 1% isoflurane with 66% nitrous oxide in oxygen. At the end of surgery, a bolus dose of remifentanil 1 microgram kg-1 (n = 20) or saline placebo (n = 20) was given and tracheal extubation was performed when standard criteria were achieved. Arterial pressure and heart rate were recorded non-invasively at 1-min intervals from the end of surgery. Remifentanil attenuated the increase in both mean arterial pressure (P < 0.001) and heart rate (P < 0.05) at extubation. Mean time to extubation was 7.2 (SEM 0.6) min and 4.0 (0.5) min in the remifentanil and saline groups, respectively (P < 0.001). There was no difference in the incidence of coughing at extubation, time to recovery from anaesthesia or time to fitness for discharge from the recovery room. PMID- 10673887 TI - Bacterial contamination of needles used for spinal and epidural anaesthesia. AB - We have investigated prospectively the incidence of bacterial contamination of 114 spinal and 20 epidural needles collected immediately after lumbar puncture of the subarachnoid or epidural space. Bacteriological examination revealed bacterial contamination of 24 (17.9%) of the needles, mainly coagulase-negative staphylococci (21; 15.7%) followed by yeasts (2; 1.5%), enterococcus (1; 0.8%), pneumococcus (1; 0.8%) and micrococcus (1; 0.8%). Our results suggest that even during aseptic puncture for lumbar anaesthesia, there is a significant rate of needle contamination. PMID- 10673888 TI - Effect of itraconazole on the pharmacokinetics of bupivacaine enantiomers in healthy volunteers. AB - We studied seven healthy volunteers given itraconazole 200 mg orally or placebo, once daily for 4 days, in a crossover study. On day 4, racemic bupivacaine 0.3 mg kg-1 was given i.v. over 60 min and venous plasma samples were collected for 23 h. Plasma concentrations of R- and S-bupivacaine, itraconazole and hydroxyitraconazole were measured. Itraconazole reduced the clearance of R bupivacaine by 21% (P < 0.05) and that of S-bupivacaine by 25% (P < 0.05), while it had no significant effect on other pharmacokinetic variables of the enantiomers. Reduction of bupivacaine clearance by itraconazole probably increases the steady-state concentration of bupivacaine enantiomers by 20-25%. This should be taken into account in the concomitant use of bupivacaine and itraconazole, although the interaction seems to be of limited clinical significance. PMID- 10673889 TI - Sensory hyperinnervation after neonatal skin wounding: effect of bupivacaine sciatic nerve block. AB - The response to tissue injury includes sensitization of peripheral nociceptors and central neuronal pathways leading to acute clinical and inflammatory pain. A further response is sprouting of sensory nerve terminals in the region of skin damage. This hyperinnervation response is particularly intense in neonates compared with adults. In this study, we tested the effect of regional nerve block at the time of injury on skin hyperinnervation. Anaesthetized newborn rat pups were treated with percutaneous sciatic nerve block injections of 0.25% bupivacaine 25 microliters followed by a localized hindpaw skin wound. Cutaneous innervation was studied by image analysis of immunostained skin sections, 7 days after wounding, and sensory thresholds were assessed using von Frey hairs. The results showed that both hyperinnervation and hypersensitivity were not significantly altered by the application of a regional nerve block at the time of injury. This suggests that regional analgesia, used commonly in clinical practice, is unlikely to prevent the hyperinnervation that follows skin wounding. PMID- 10673890 TI - Propofol anaesthesia in mice is potentiated by muscimol and reversed by bicuculline. AB - We have examined the role of gamma-aminobutyric acid (GABA) neurones in propofol anaesthesia in mice using the righting reflex. Propofol i.p. increased the percentage of loss of the righting reflex in a dose-dependent manner with an ED50 value of 140 (95% confidence limits 123-160) mg kg-1 (n = 40; eight animals per dose, five doses per dose-response curve). The ED50 for propofol decreased significantly to 66 (58-75) mg kg-1 in the presence of the GABAA receptor agonist muscimol 1 mg kg-1 i.p. (n = 40) (P < 0.05). In contrast, the ED50 increased significantly to 240 (211-274) mg kg-1 in the presence of the antagonist bicuculline 5 mg kg-1 i.p. (n = 40) (P < 0.05). Our results suggest that propofol anaesthesia may be mediated, at least in part by GABA neurons. PMID- 10673891 TI - Pulmonary haemorrhage after percutaneous paravertebral block. AB - We describe the management of a 65-yr-old woman anaesthetized for thoracotomy. The patient suffered a pulmonary haemorrhage after percutaneous paravertebral injection performed using the loss of resistance to saline technique. Thoracotomy at a later date revealed that the lung tissue had become adherent to the chest wall and that the paravertebral space was fibrosed secondary to previous surgery. This particular complication of percutaneous paravertebral block has not been reported previously and raises the question of risk vs benefit of this pre emptive analgesic technique. PMID- 10673892 TI - Combined spinal-epidural in the obstetric patient with Harrington rods assisted by ultrasonography. AB - We describe a patient with severe scoliosis, which had been corrected partially with Harrington rods, who requested epidural analgesia for labour. With no palpable landmarks, the use of ultrasound enabled identification of the vertebral midline and allowed provision of regional anaesthesia. PMID- 10673893 TI - Ropivacaine and bupivacaine for long-term epidural infusion in a small child. AB - Ropivacaine is assumed to be less toxic than bupivacaine but there are no reports concerning its long-term use in paediatric anaesthesia. We report the use of ropivacaine for long-term epidural anaesthesia in a 21-month-old girl. In two consecutive periods of 3 days each, 0.5% bupivacaine and 0.5% or 0.75% ropivacaine were administered to facilitate painful vaginal brachytherapy. The mean dose of bupivacaine increased from 1.05 to 1.32 mg kg-1 h-1 and that of ropivacaine increased from 1.40 to 3.86 mg kg-1 h-1. No toxic side effects were observed. We conclude that both epidural ropivacaine and bupivacaine were effective and safe during long-term epidural anaesthesia in this particular case. However, the doses were potentially toxic and should therefore be used with extreme caution. PMID- 10673894 TI - A rough guide to molecular biology. PMID- 10673895 TI - Hypoxia caused by a faulty Steri-cath closed suction device and use of continuous suction. PMID- 10673896 TI - Placement of double-lumen endobronchial tubes. PMID- 10673897 TI - Fibreoptic orotracheal intubation. PMID- 10673898 TI - Is anaesthesia evidence-based? PMID- 10673899 TI - Do not rely on a laryngeal mask in major periglottic pathology. PMID- 10673900 TI - EEG indices and heart rate variability as measures of depth of anaesthesia. PMID- 10673901 TI - Regional anaesthesia for carotid endarterectomy. PMID- 10673902 TI - Regional anaesthesia for carotid endarterectomy. PMID- 10673903 TI - Central nerve block and thromboprophylaxis. PMID- 10673904 TI - Human milk oligosaccharides: 130 reasons to breast-feed. PMID- 10673905 TI - Dietary fatty acids and atherosclerosis regression. PMID- 10673906 TI - The control of partitioning between protein and fat during human starvation: its internal determinants and biological significance. AB - Human subjects vary in the extent to which their body's protein and fat compartments are mobilized for fuel during starvation. Although an inverse association between the initial adiposity and the contribution of protein as fuel during starvation has been known for nearly a century, interest in the quantitative importance and functional significance of the initial percentage fat as a determinant of biological variation in energy-partitioning between protein and fat (and hence in determining the partitioning characteristic of the individual) is relatively recent. The present paper addresses these issues by revisiting the classic Minnesota experiment of semi-starvation and refeeding from a standpoint of system physiology. In a quantitative analysis of the relationship between the initial body composition (ration FAT0: fat-free mass (FFM)0) and the composition of weight loss (ratio delta FAT: delta FFM) in the thirty-two men in the Minnesota study, the arguments are put forward that the fraction of FFM lost when the fat stores reach total depletion is independent of the initial percentage fat, and that this fraction represents the 'dispensable' component of the protein compartment that is compatible with life (i.e. the protein energy reserve, rp). The concepts are developed that (1) the initial percentage body fat (which reflects the initial ratio FAT0:FFM0) provides a 'memory of partitioning' which dictates the control of partitioning between protein and fat in such a way that both the protein energy-reserve (rp) and the fat energy-reserve (rf) each complete depletion simultaneously, a strategy that would ensure maximum length of survival during long-term food scarcity, and that (2) variability in the relative sizes of these two energy reserves (i.e. in rf:rp) could, in addition to the initial percentage fat, also contribute to human variability in energy partitioning. The basic assumptions underlying this re-analysis of the Minnesota data, and the concepts that are derived from it, have been integrated in the simple mathematical model for predicting the partitioning characteristic of the individual. This model is used to explain how variability in the fraction of the protein compartment that could function as an energy reserve (rp) can be as important as the initial percentage fat in determining inter-individual variability in protein-sparing during the early phase of starvation, in fuel partitioning during prolonged starvation, or in the maximum percentage weight loss during starvation. The elucidation of factors underlying variability in the size of the protein energy-reserve may have important implications for our understanding of the pathophysiology of starvation and age-associated susceptibility to muscle wasting, and in the clinical management of cachexia and obesity. PMID- 10673907 TI - Selenium supplementation affects the retention of stable isotopes of selenium in human subjects consuming diets low in selenium. AB - Twenty-nine women and fifteen men from an area of low Se intake (South Island of New Zealand) consumed 100 micrograms stable 74Se, as selenate given in water after an overnight fast, and blood was collected for 3 weeks. They were then divided into five groups and supplemented with 0, 10, 20, 30 and 40 micrograms Se/d (as selenomethionine) for 5 months. After 5 months, they received a second dose of 74Se identical to the first. Supplementation significantly altered retention of 74Se in the plasma, but not in the erythrocytes or platelets. Subjects receiving the placebo retained the greatest amount, and subjects receiving 30 micrograms supplemental Se/d retained the least 74Se. Supplementation resulted in relatively more isotope being retained in a medium molecular mass protein considered to be albumin, and relatively less in another fraction considered to be selenoprotein P. The lack of many observed changes in retention of stable Se, and the shift in retention among the plasma proteins, suggests that supplemental Se was not being used to replete critical pools of Se, probably because of adaptation to low Se intake. PMID- 10673908 TI - Neutral oligosaccharide content of preterm human milk. AB - Human milk oligosaccharides are known to play a role in protection against certain infectious diseases. Previous reports indicate that the content of human milk oligosaccharides varies widely among individuals at term but such information on preterm milk is lacking. After removal of the fat, protein and most of the lactose from non-pooled human milk samples, a total neutral oligosaccharide fraction was isolated by ion-exchange chromatography followed by gel filtration. A Dionex high-performance anion-exchange chromatography system equipped with a pulsed electrometric detector was then employed to measure the levels of ten neutral oligosaccharides in the individual milk samples. Twenty three milk samples from thirteen mothers who delivered at a mean gestational age of 29.5 (SD 3.1) weeks were collected between days 0 and 33 of lactation, and compared with three samples of term milk from two mothers. The ranges of the total and individual levels of the ten neutral oligosaccharides in preterm milk were similar to those in term milk. Further, as previously described in term milk, preterm milk exhibited a quantitative individual variation. This variation was independent of the gestational age, day of lactation, and postconceptional age. In conclusion, levels of ten neutral oligosaccharides did not differ between preterm and term human milk. PMID- 10673909 TI - Dietary and socio-economic factors associated with obesity among Kuwaiti college men. AB - Obesity has been on the increase among people of the Arabian Gulf countries. Overweight and obesity among 18-29-year-old Kuwaiti men increased by 23.4 and 14.8% respectively, between 1980 and 1993. The objective of the present study was to explore factors associated with overweight and obesity in a sample of 515 Kuwaiti college men studied in 1997. Weight and height were measured. The index of adiposity used was the BMI, which is the weight (kg) divided by the height (m) squared (kg/m2). The men were classified as overweight (BMI > 25 kg/m2) or obese (BMI > 30 kg/m2). The associated factors obtained through questionnaires included age, marital status, governorate, number of siblings, suffering from a chronic disease, subjects' parental obesity, education and occupation, number of major meals eaten, eating between meals, family income, number of servants, number of people living at home, exercising, last dental and physical check-up, dieting, year of study, highest desired degree after college, countries preferred for visiting, and socio-economic status. The results of the study revealed that 38.5 and 11.1% of the students were overweight and obese respectively. Factors that were found to be significantly associated with overweight and obesity among the men included age, marital status, last dental check-up, exercising, subjects' parental obesity, dieting and year of study. Logistic regression analysis of significant associated factors revealed that the same factors contributed to the development of overweight and obesity. PMID- 10673910 TI - Effects of inulin on faecal bifidobacteria in human subjects. AB - A controlled study with eight healthy free-living subjects was carried out, in which energy intake was adjusted to the individual energy requirements. On administration of inulin, blood lipids, the faecal microflora, short-chain fatty acids and accompanying gastrointestinal symptoms were characterized in order to investigate the long-term effect of inulin. During the run-in phase (8 d), subjects received a typical Western diet providing 45% energy as fat and 40% energy as carbohydrate. Subsequently, the subjects consumed a fat-reduced diet which provided 30% energy as fat and 55% energy as carbohydrate for a period of 64 d using inulin as a fat replacer. The amounts of inulin consumed by the subjects (up to 34 g/d) were based on individual energy requirements with the aim to keep the diet isoenergetic with that used in the run-in period. To assess the effects of inulin administration, a control study (run-in and intervention) was carried out in which subjects consumed the same diet but devoid of inulin during the whole course of the study. To investigate the effect of inulin on faecal flora composition total bacteria and bifidobacteria in the faeces were enumerated by in situ hybridization with 16S rRNA targeted oligonucleotide probes. Inulin significantly increased bifidobacteria from 9.8 to 11.0 log10/g dry faeces and caused a moderate increase in gastrointestinal symptoms such as flatulence and bloatedness, whereas blood lipids and short-chain fatty acids remained essentially unaffected. PMID- 10673911 TI - Modulation of human T-lymphocyte functions by the consumption of carotenoid-rich vegetables. AB - A human intervention study was conducted to determine the effect of the consumption of carotenoid-rich vegetables on the immune system. Subjects, (twenty three men), who were non-smokers, were not restricted in their daily diet, except that they had to abstain from fruit and vegetables high in carotenoids throughout the whole study period. The study was divided into four periods, each lasting 2 weeks: weeks 1-2: low-carotenoid period; throughout weeks 3-8: daily consumption of 330 ml tomato juice (40 mg lycopene/d, 1.5 mg beta-carotene/d) (weeks 3-4), 330 ml carrot juice (21.6 mg beta-carotene/d, 15.7 mg alpha-carotene/d, 0.5 mg lutein/d) (weeks 5-6), 10 g dried spinach powder (11.3 mg lutein/d, 3.1 mg beta carotene/d) (weeks 7-8). Blood was collected weekly from subjects after a 12 h fast. T-lymphocyte functions were assessed by measuring proliferation and secretion of immunoreactive cytokines. The consumption of a low-carotenoid diet resulted in a significantly reduced proliferation of peripheral blood mononuclear cells (PBMC) cultured with concanavalin A. After 2 weeks of tomato juice consumption and until the end of the intervention period lymphocyte proliferation was not significantly changed compared with proliferation at the end of the depletion period. Secretion of cytokines by T-helper-1-like lymphocytes (interleukin (IL)-2) and by T-helper-2-like lymphocytes (IL-4) was influenced by the dietary intervention. IL-2 and IL-4 secretion values were significantly suppressed after the low-carotenoid diet (P < 0.001 and P < 0.05 respectively compared with baseline). Tomato juice consumption significantly enhanced IL-2 (P < 0.001) and IL-4 secretion (P < 0.05) compared with the end of depletion period. After carrot juice and spinach powder consumption the cytokine secretion capacity of PBMC was not significantly different from that at the end of the depletion period. In conclusion, the results of the present study indicate that a low carotenoid diet reduces T-lymphocyte functions and addition of tomato juice restores these functions. This modulation could not be explained by changes in the plasma carotenoid concentrations. The active constituents in tomato juice as well as the biological significance of this immunomodulation remain to be determined. PMID- 10673912 TI - Lactose-derived oligosaccharides in the milk of elephants: comparison with human milk. AB - Human milk is commonly considered to be unique when compared with the milk of other species with regard to its high content of complex fucosylated and sialylated lactose-derived oligosaccharides. We describe the application of high pH anion-exchange chromatography with pulsed amperometric detection and TLC to characterize and quantitate neutral and sialylated lactose-derived oligosaccharides in milk from three Asian elephants and human milk. The lactose contents of elephant and human milks were 25-30 g/l and about 66 g/l respectively, whereas total oligosaccharide concentration was about three times higher in elephant milk and comprised up to 40% (10% in human milk) of the carbohydrate content. The ratio neutral: acidic components was different in the milk of the two species; in elephant milk, the N-acetylneuraminic acid-containing oligosaccharides made up almost half of the total amount v. 30% in human milk. Most oligosaccharides in elephant milk were more fucosylated and/or sialylated compared with human milk components. By mild acid hydrolysis, fucose and N acetylneuraminic acid were cleaved off from complex components, and this resulted in increased amounts of fucose, galactose, N-acetylneuraminic acid, lactose and lacto-N-neo-tetraose. Unique to elephant milk are the high levels of 3' galactosyllactose (up to 4 g/l) and lacto-N-neo-tetraose which are present in human milk only in trace amounts. Elephant and human milks have high levels and unique patterns of oligosaccharides which may reflect the relative importance of these components in neonatal host defence, in endothelial leucocyte interactions or in brain development. PMID- 10673914 TI - Phospholipid fatty acid composition and protein kinase C activity in the large intestine of rats fed on butter and coconut-oil diets. AB - Protein kinase C (PKC) has been proposed to play an important role in the aetiology of colon cancer. Therefore, we investigated whether the amount and type of saturated fat could affect colonic PKC activity by modifying either mucosal phospholipid fatty acid composition or faecal diacylglycerol production. Male Wistar rats (n 13 per group) were fed on diets containing butter or coconut oil at energy levels of 10% and 43% for 4 weeks. The control group received a low-fat diet providing 10% of energy from sunflowerseed oil. PKC activity was higher in the distal than the proximal colon but the quantity or type of fat did not alter PKC activity in either region of the colon. Saturated fats caused moderate changes in the fatty acid composition of caecal phospholipids, which were more obvious in the phosphatidylethanolamine than in the phosphatidylcholine fraction. A significant correlation was found between fatty acid composition of phosphatidylcholine and membrane PKC activity. In particular, there was a positive correlation between the proportion of saturated 14:0 and 18:0 and increased PKC activity while unsaturated 18:2n-6, 20:4n-6 and 16:1n-7 were inversely correlated with PKC activity. No relationship was found between phosphatidylethanolamine fatty acids and PKC activity. Concentration of faecal diacylglycerol was not affected by the diet. Overall the data suggest that diets high in saturated fat may not alter colonic PKC activity to a significant extent. PMID- 10673913 TI - Modulation of the regression of atherosclerosis in the hamster by dietary lipids: comparison of coconut oil and olive oil. AB - The Golden Syrian hamster (Mesocricetus auratus) has been shown to be a useful model of both human lipoprotein metabolism and the development of atherosclerosis. We report the effects of dietary lipids on the progression and regression of atherosclerosis in this model. In the first study, hamsters fed on coconut oil (150 g/kg diet) and cholesterol (30 g/kg diet) developed lipid-rich lesions in the ascending aorta (0.28 (SD 0.14) mm2) and aortic arch (0.01 (SD 0.01) mm2) after 4 weeks that continued to progress over the next 8 weeks (0.75 (SD 0.41) mm2 and 0.12 (SD 0.11) mm2 for the ascending aorta and aortic arch respectively). Removal of cholesterol from the diet halted this progression. Furthermore, in animals fed on olive oil in the absence of added cholesterol, plasma LDL-cholesterol concentrations were lower (P < 0.05) and the extent of atherosclerotic lesions was reduced (P < 0.001 for both regions of the aorta) compared with animals fed on coconut oil (with no added cholesterol). In a second study, animals were fed on the atherogenic diet for 10 weeks, transferred to diets containing either coconut oil (150 g/kg diet) or olive oil (150 g/kg diet) without added cholesterol and monitored for up to 16 weeks. In the ascending aorta, lesion size doubled in animals fed on coconut oil but stabilized in those fed on olive oil. In the aortic arch, lesion size decreased linearly (P < 0.05, P < 0.001 for coconut oil and olive oil respectively) with the greatest reduction being seen in the olive-oil-fed animals (P < 0.05). Again, progression and regression of atherosclerosis appeared to reflect the relative concentrations of LDL-cholesterol and HDL-cholesterol in the plasma. We conclude that the male Golden Syrian hamster represents a useful model of dietary induced regression as well as progression of atherosclerosis. PMID- 10673915 TI - Enhancement of butyrate production in the rat caecocolonic tract by long-term ingestion of resistant potato starch. AB - Some data suggest that the colonic microflora may adapt to produce more butyrate if given time and the proper substrate. To test this hypothesis, we investigated the effect of prolonged feeding of resistant potato starch on butyrate production. Rats were fed on either a low-fibre diet (basal) or the same diet supplemented with 90 g resistant potato starch/kg (PoS) for 0.5, 2 and 6 months. Short-chain fatty acid (SCFA) concentrations were determined in caecal and colonic contents at the end of each ingestion period. Total SCFA concentration increased over time throughout the caecocolonic tract with PoS, but was not modified with the basal diet. While propionate concentration was unchanged, butyrate concentration was highly increased by PoS at each time period in both the caecum and colon. Moreover, the butyrogenic effect of PoS increased over time, and the amount of butyrate was increased 6-fold in the caecum and proximal colon and 3-fold in the distal colon after 6 months compared with 0.5 months. Accordingly, the ratio butyrate:- total SCFA increased over time throughout the caecocolonic tract (12.6 (SE 2.8) v. 28 (SE 1.8)% in the caecum, 10.5 (SE 1.4) v. 26.8 (SE 0.9)% in the proximal colon, and 7.3 (SE 2.4) v. 23.9 (SE 2.7)% in the distal colon at 0.5 v. 6 months respectively), while the proportion of acetate decreased. Neither the proportion nor the concentration of butyrate was modified over time with the basal diet. Butyrate production was thus promoted by long-term ingestion of PoS, from the caecum towards the distal colon, which suggests that a slow adaptive process occurs within the digestive tract in response to a chronic load of indigestible carbohydrates. PMID- 10673916 TI - Oromandibular reconstruction using a third free flap in sequence in recurrent carcinoma. AB - Successful results of a second microsurgical reconstructive attempt have been reported previously in recurrent oral carcinoma. However, the feasibility of a third free flap following a third excision has remained to be determined. Six oral carcinoma patients with multiple recurrences, surgical excisions and free flap reconstructions on three separate occasions are presented. Resections had a curative intent in all cases in the first and second ablations and in four of the six cases in the third one. Five radial forearm flaps and one double free flap were used for the first reconstruction. During the second reconstruction two radial forearm, two fibula osteoseptocutaneous, one double free flap and one rectus femoris flap were used to reconstruct the larger defects resulting from excision of the recurrent tumours. However, no vascularised bone transfers were performed following the third excision and soft tissue free flap plus plate option was used for segmental mandibular defects. There was one partial flap loss among 21 free flaps performed. Three patients died within an average of 8 months following the third reconstruction while the others remained alive, surviving an average of 6 months. In conclusion, a third free tissue transfer for reconstruction in multiply recurrent oral carcinoma was found to be feasible, safe and effective. The use of free flaps contributed to the prevention of complications in these difficult cases and enabled the patients to spend the remaining days of their lives outside hospital. PMID- 10673917 TI - Expression of growth factors in the mandibular distraction zone: a sheep study. AB - Interest in craniofacial osteodistraction has increased in recent years parallel with the growing attention given to the role of growth factors in tissue healing and regeneration. This study was embarked upon to investigate the expression of bFGF, TGF-beta and IGF-1 in the distraction zone of the mandible. Fourteen growing sheep were allocated to three experimental groups. Six animals were allocated to Groups A and B (n = 12) and underwent bilateral mandibular corticotomies with fixation of an external lengthening device. The distraction protocol consisted of a rate of 1.0 mm/day (twice daily) for 20 days followed by a consolidation phase of 20 days after which the sheep were sacrificed. Group C comprised of age matched sham operated animals (n = 2). Bone histochemistry for growth factors were performed in the harvested mandibles. A strong staining of bFGF was seen in the osteoblasts, osteocytes and osteoid matrix following 20 days of distraction and 20 days of consolidation compared to the control group. TGF beta and IGF-1 demonstrated mild but clear staining in osteocyte and osteoblast cells and TGF-beta stained positively in the osteoid seam in the experimental groups. These finding suggest that bFGF, IGF-1 and TGF-beta may play different roles in the remodelling phase of distraction osteogenesis. PMID- 10673918 TI - Analysis of flow changes in forearm arteries after raising the radial forearm flap: a prospective study using colour duplex imaging. AB - The purpose of this study is to assess the changes in flow patterns of forearm arteries produced by excision of the radial artery when harvesting the radial forearm flap, in order to clarify its vascular morbidity rationally. Eleven patients with elective surgery using the radial flap were included in this investigation. A prospective study was designed using colour duplex imaging for quantitative flow measurement in two stages: a few days before the operation, a first colour duplex scanning examination was done recording flow velocity and vessel section area from the radial, ulnar, posterior interosseous and anterior interosseous arteries around the wrist. Volumetric parameters and relative blood flow percentages were calculated and compared to those obtained from a second similar vascular investigation accomplished in the same limb 4-5 months after the operation. Statistical analysis was done using the Wilcoxon matched pairs test. After raising the radial forearm flap there was a trend for increased overall forearm flow (from 162 to 215 ml/min, P = 0.09 N.S.), the ulnar (P = 0.04), the posterior interosseous (P = 0.004) and the anterior interosseous (P = 0.003) arteries being responsible for this tendency. The anterior interosseous artery showed the greatest increase in blood (from 8.2 to 67.7 ml/min), reaching a relative flow percentage (33%) close to that of the radial artery before its excision (39%). Results of this study indicate that another 'major vascular axis' based on the anterior interosseous artery develops after sacrificing the radial artery and that global arterial inflow to the hand is not impaired. PMID- 10673919 TI - Reduction of potential contamination of breast implants by the use of 'nipple shields'. AB - Forty-three breast implant operations in 25 patients were studied prospectively to determine the effectiveness of covering the nipple-areolar complex with an adhesive film dressing in preventing perioperative expression of bacteria from nipple ducts contaminating the operative field. One swab from the nipple after skin preparation and none of the swabs taken from the outer surface of the film dressing postoperatively yielded any bacterial growth. Fourteen breasts (33%) in 11 patients (44%) yielded bacterial growth from swabs under the film postoperatively. Six of 9 breasts (67%) in 5 patients who had capsulectomies had bacteria isolated from under the film postoperatively. Ten of 14 (71%) control breasts (no shields) in 6 of 7 patients (86%) had positive postoperative swabs. This study confirms the potential risk of bacterial contamination arising from nipple duct flora during intra-operative breast manipulation, and the effectiveness of a perioperative adhesive film placed over the nipple-areolar complex in preventing subclinical bacterial contamination of implanted breast prostheses. PMID- 10673920 TI - Psychosocial outcome and patient satisfaction following breast reduction surgery. AB - There is an increasing awareness that psychosocial outcome and health status are important outcomes following breast reduction surgery. In this study, patients awaiting breast reduction surgery completed detailed and comprehensive psychosocial assessments before and after surgery. Of 33 patients who completed the preoperative assessment, 20 patients were operated on and 19 were reassessed 4 months post-surgery. Patients expressed high levels of satisfaction with specific and overall results of surgery. Scores for anxiety, depression, body image and body satisfaction improved significantly using specific questionnaires. Patients also reported significant improvements on five out of eight subscales on the Short Form 36 health status questionnaire. This study provides further evidence for overall improvement in health status and psychological functioning in patients undergoing breast reduction surgery and supports the provision of this service by the NHS. PMID- 10673921 TI - Preliminary report: the distally pedicled dorsoulnar forearm flap for hand reconstruction. AB - In this paper, a new flap for the coverage of soft tissue defects in the hand is described. To obtain a distally pedicled dorsoulnar flap, the dissection of the standard dorsoulnar flap is continued distally under the descending branch of the dorsal branch of the ulnar artery onto the dorsum of the wrist after the dorsal branch of the ulnar artery is ligated and divided. Our modification of the standard dorsoulnar flap converts this flap to a distally based flap, which provides a potentially longer pedicle and increases the arc of rotation of the flap. Two successful cases are reported. PMID- 10673922 TI - Closure of elective skin defects in the leg with a fasciocutaneous V-Y island flap. AB - Skin defects in the leg can be among the most difficult to cover. Split skin grafting of such defects can be complicated by delayed healing and poor cosmesis and this has led to the description of several local flap techniques. Here a technique of V-Y flap closure is described based laterally on a random fascial pedicle. Twenty patients aged from 51 to 85 years had flaps for skin closure after excision of skin tumours, 17 of them in the middle or lower third of the leg. All but one were performed under local anaesthetic, seven as day cases and eight with an overnight stay. No patient was excluded because of medical conditions or skin quality and two patients had significant lymphoedema. A single V-Y flap was used in each case and was found to have much greater mobility than previously described similar flaps. Cosmetic results, and in particular contour preservation, were found to be excellent. There were no complete flap losses but four patients had delayed healing, two related to partial flap necrosis. Four patients had wound infections, one requiring readmission for intravenous antibiotics but no patient had further surgery. The fasciocutaneous V to Y island flap was found to be a very satisfactory method of wound closure in the leg with an acceptable complication rate and excellent cosmetic results. PMID- 10673923 TI - Emergency management of type IIIB open tibial fractures. AB - We present our therapeutic strategy for the treatment of type IIIB open tibial fractures. It involves emergency internal stabilisation of the bone by locked intra-medullary nailing when appropriate and skin cover using either a pedicled or free muscle flap. Where there is bone loss, a cancellous iliac graft is performed at the same time. Eighteen cases of type IIIB open tibial fractures treated between 1986 and 1995 were analysed. There were 17 men and 1 woman; the average age was 35 years. Each of the 18 patients underwent wound debridement as a primary emergency procedure with no secondary reoperation. Bone fixation was performed by locked intra-medullary nailing (AO nail, How Medica) 6-10 h after trauma. A primary cancellous iliac bone graft was performed in three cases. Cover was applied immediately after nailing (muscular pedicle flaps in 12 cases, muscular free flaps in 6 cases). Local flap cover led to two failures: both these fractures were followed by postoperative complications. The 6 free muscle flaps were successful. The average time to bone union was 6.5 months (range: 3-18.5 months) according to clinical criteria and 9 months (range: 4-27 months) according to radiological criteria. Out of the 18 fractures, 13 were primarily united (72.2% of cases); 3 involved osteitis and 2 nonunion. Sixteen patients were examined again with a mean follow-up of 4.8 years (range: 1-11 years). Six moderate malunions occurred; none needed surgical reoperation. Ankle motion was normal in 7 cases and reduced to below 50% in 9 cases when compared with the healthy ankle. Thirteen patients resumed their previous professional activities. This surgical strategy reduces bone union time, the number of operations and the time spent in hospital; it improves functional results. PMID- 10673924 TI - Reconstruction of perianal skin defect using a V-Y advancement of bilateral gluteus maximus musculocutaneous flaps: reconstruction considering anal cleft and anal function. AB - In order to preserve the anal function after ano-perianal skin excision for malignancy, we have reconstructed a deep, symmetrical natal cleft using a V-Y advancement of bilateral gluteus maximus musculocutaneous flaps thinned medially and sutured to the ooccyx, anococcygeal ligament and the central tendon of the perineum. This technique was applied in three cases of Bowen's disease and two cases of Paget's disease. In all five cases, postoperative anal functions such as comfortable defecation and sensation, were well preserved, the perianal skin and underwear stayed clean, and there was no disturbance of walking or exercise. PMID- 10673925 TI - Recurrence rates of ischial sores in para- and tetraplegics treated with hamstring flaps: an 8-year study. AB - We have collected data on the second follow-up of 27 patients who underwent musculocutaneous flap closure of their ischial pressure sores. Thirty-seven ulcers were operated on between 1988 and 1993 using the V-Y advancement hamstring musculocutaneous island flap. At the initial follow-up (mean = 20 months) in 1993, despite 33% of patients having had recurrent ulcers and 14.8% having undergone re-advancements, only 14% of patients had non-healing ulcers. In 1997, follow-up period ranged from 18 to 90 months, with a mean of 62 months. Four patients were lost to follow-up resulting in 23 patients (n = 23) for the current study. Nine patients were tetraplegic and the remaining 14 were paraplegic. Four of the 23 patients had died at follow-up therefore making the number of living patients 19 (n = 19). The total number of ulcers operated on in the current study was 29 (U = 29). Overall, ulcer and patient recurrence rates were 41.4% and 47.8% respectively. Despite this, 89.5% of patients had intact flaps at the time of follow-up. We recommend the use of the hamstring V-Y musculocutaneous flap as a reliable and safe reconstructive modality in the management of ischial pressure sores and by identifying the group of patients susceptible to ulcer recurrence we have proposed a protocol for their long-term follow-up. PMID- 10673926 TI - Subcutaneous adrenaline infiltration in paediatric burn surgery. AB - Paediatric burn surgery may be associated with significant blood loss and postoperative pain. To investigate methods of reducing these symptoms, we studied a prospective series of 29 children with small to medium sized burns. Presurgically both the burn wound and split skin graft donor sites were injected with a 1:500,000 adrenaline solution, to which bupivicaine had been added. No patient required blood transfusion and no patient developed systemic side effects from the injected solution. Four patients required parenteral analgesia, two in the immediate postoperative period and two at first dressing change. In all other patients pain was controlled with oral analgesia alone. Mean graft take in our series was 95%, indicating that this technique does not compromise burn depth assessment, nor impair graft survival. PMID- 10673927 TI - Colour shift following tattoo removal with Q-switched Nd-YAG laser (1064/532). AB - Colour shift in tattoo pigment is a recognised complication of laser tattoo treatment. We report our experience over the past 4 years in treating 275 patients, with a total of 323 professional tattoos. Of these, 184 tattoos contained a pigment other than black with 33 displaying a colour shift as a consequence of laser treatment. This adverse effect was recorded with red, yellow, crimson and brown pigments but most frequently with white/flesh pigments. We found brown and white/flesh coloured pigments to be significantly more likely to change colour compared to red and that the chance of certain colours shifting related to the age of the tattoo. We outline our management of this problem and discuss the results of continued treatment. PMID- 10673928 TI - The establishment of a burns unit in a developing country--a collaborative venture in Malawi. AB - In September 1993, the recently completed 32-bedded Burns Unit in the Queen Elizabeth Central Hospital, Blantyre, Malawi, was officially opened. This represented the culmination of 3 years' planning and construction; this paper presents our experience in developing this type of project, and an analysis of the early results of treatment in the unit. The widespread neglect of this common injury in developing countries is highlighted and the importance of input from specialists in this field working in collaboration with local medical and nursing staff is emphasized. The potential for making a significant impact in reducing the morbidity and mortality in burn injuries is highlighted. PMID- 10673929 TI - Digital surgery revisited. On the origin of digital transfer in reconstructive surgery. AB - A scarred donor site has been of secondary concern in reconstructive surgery, being the price one had to pay for the repair of a defect. In the 19th century reconstruction at the sacrifice of a little finger was contemplated and applied to difficult nasal repairs. The loss of a finger was sometimes preferred thus avoiding additional scars on the forehead and cheek. This so called 'Russian method' is of British origin, described in 1875 by James Hardie of Manchester. PMID- 10673930 TI - Pseudocyst formation after abdominoplasty--extravasations of Morel-Lavallee. AB - A soft tissue injury can lead to the formation of a pseudocyst in the subcutaneous adipose tissue, due to a seroma, haematoma or fat necrosis. These cysts were first described in 1853 by the French physician Morel-Lavallee. He observed the phenomenon in the lower limb in women after a tangential trauma with separation of the fatty layers. A similar condition can occur following surgery of the abdomen, when performing liposuction and subcutaneous dissection with a large dead space. In this report we present two cases with large pseudocysts in the abdominal wall, which were seen in long term follow-up after an abdominoplasty performed elsewhere. The pathogenesis, the treatment and the literature are discussed. PMID- 10673931 TI - Microneurovascular transfer of contralateral latissimus dorsi in Poland's syndrome. AB - A 2-year-old girl with Poland's syndrome presented with syndactyly of the first, second and third webs of the left hand and ipsilateral absence of the sternocostal portion of pectoralis major. Latissimus dorsi on the same side was also absent. A microvascular free contralateral latissimus dorsi muscle transfer was undertaken to reconstruct the chest wall and anterior axillary fold. The transfer was innervated by intercostal nerve transfer and produced a neuromuscular unit of good aesthetic appearance and function. PMID- 10673932 TI - Subcutaneous emphysema of a digit through a pre-existing puncture wound. AB - A case of injection of compressed air into a digit is reported. The air was injected at 50 PSI through a trivial puncture wound sustained some hours previously. The case had a benign course, in comparison to high pressure injection injuries with foreign material. PMID- 10673933 TI - An unusual case of orbital trauma with a large foreign body in the maxilla. AB - A case in which a broken handle of a scooter was found lodged in the right maxillary sinus and infratemporal fossa with an unusual route of entry through the orbital floor without any injury to the globe is reported. PMID- 10673934 TI - Experience of methicillin-resistant Staphylococcus aureus in a plastic surgery unit. PMID- 10673935 TI - The use of blood as a marker on the lips: an aid to reconstruction--reply. PMID- 10673936 TI - A simple method of decreasing the morbidity of split-thickness skin graft donor sites. PMID- 10673937 TI - Breast reduction--rationed or rational? PMID- 10673938 TI - Placement of sutures in tendon repair. PMID- 10673939 TI - An efficient form of note taking. PMID- 10673940 TI - Trilucent breast implants--a personal review of the current controversies (May 1999) PMID- 10673941 TI - The cancer patient in the radiology department: do we live up to our responsibilities? PMID- 10673942 TI - Features of unusual metastases from prostate cancer. AB - A retrospective study of CT examinations on 508 patients with prostate carcinoma was performed in order to document the presence or absence of uncommon nodal and extranodal metastases according to pre-defined criteria. 45 atypical metastases were demonstrated in 36 patients. There were 23 atypical nodal metastases including unusual distribution (19/23), large volume (7/23) and atypical morphology or contrast enhancement pattern (8/23). 22 atypical extranodal metastases included orbit/skull base (11), pulmonary (4), liver (3), intracranial (2), ocular (1) and adrenal (1). Atypical prostate carcinoma metastases are usually encountered in the presence of known advanced disease but can be the presenting feature of malignancy or the only sign of distant spread. Knowledge of atypical manifestations of metastatic disease will reduce diagnostic delay, allow accurate staging and lead to the effective and timely delivery of appropriate treatment. PMID- 10673943 TI - Ultrasound of testicular epidermoid cysts. AB - The ultrasound findings of focal intratesticular lesions may sometimes be characteristic of benign pathology, which accounts for approximately 5% of all testicular tumours. Three cases of epidermoid cysts of the testis are presented along with a review of the literature. Recognition of the typical ultrasound appearances of this entity in a clinical situation of painless testicular swelling in the absence of a history of trauma and signs of inflammation can make testis-sparing surgery feasible. PMID- 10673944 TI - First trimester nuchal translucency: effective routine screening for Down's syndrome. AB - The objective was to evaluate the effectiveness of 10-14 week nuchal translucency measurement in routine ultrasound screening for Down's syndrome. 11,398 women were scanned at 10-14 weeks of pregnancy for nuchal translucency measurements. The mean maternal age of the screened population was not significantly different from that of the booking population. A 5% screen positive rate was achieved by using a nuchal translucency-derived risk of > or = 1:200. Screening using this nuchal translucency risk would enable the first trimester detection of 16 out of 21 (76%) fetuses with Down's syndrome and 40 out of 49 (81%) aneuploid fetuses. In conclusion, this study demonstrates that first trimester nuchal translucency measurement is an effective method of screening for fetal chromosomal abnormality. PMID- 10673945 TI - Abdominal ultrasound in acute schistosomiasis mansoni. AB - Reports on abdominal ultrasound studies in patients with acute schistosomiasis are still scarce and limited data are available on structural changes of the liver parenchyma in this stage of the disease. 26 patients with acute schistosomiasis mansoni were submitted to clinical and ultrasound examination. For ultrasound comparison, each acute patient was paired by age, gender, weight and height to a non-infected individual. Ultrasound showed a non-specific homogeneous size increase of the liver, and spleen in all acute patients, and easily identified intraabdominal lymph nodes in the periportal region in most cases. Three out of the five patients with periportal thickening underwent percutaneous liver biopsy. Periportal thickening disappeared 6 months after treatment for schistosomiasis. 24 months after successful treatment there was involution of the liver and spleen; lymph nodes, although reduced in size, were still easily recognized. Liver biopsy showed dense inflammatory infiltration of neutrophils, macrophages and eosinophils in the portal tracts associated with discrete fibrous tissue formation. PMID- 10673946 TI - Ultrasound guided fine needle aspiration biopsy of splenic lesions. AB - Fine needle aspiration biopsy (FNAB) of focal splenic lesions has been infrequently utilized because of the risk of haemorrhage. This study was carried out to evaluate the safety and efficacy of ultrasound guided FNAB of splenic lesions. 35 patients with focal splenic lesions underwent FNAB under real-time ultrasound guidance using a free hand technique. Ultrasound findings were single or multiple focal hypoechoic lesions (n = 33), focal hyperechoic lesion (n = 1) and diffuse heterogeneous echotexture (n = 1). Aspirations were performed with 22 G spinal needles using either the subcostal or the intercostal approach. Definite cytological diagnosis was made in 22 patients (62.8%), including tuberculosis in 10 patients, lymphoma in seven patients, extramedullary haematopoiesis in two patients and aspergillosis, histoplasmosis and bacterial abscess in one patient each. FNAB was negative in 12 patients because the aspirates were either scanty or contained only blood. FNAB was falsely positive in one patient. Only one patient had significant intraabdominal bleeding, which was managed conservatively. In conclusion, splenic FNAB performed under ultrasound guidance is a safe and accurate method in the diagnosis of focal splenic lesions. PMID- 10673947 TI - Measurement imprecision in vertebral morphometry of spinal radiographs obtained in the European Prospective Osteoporosis Study: consequences for the identification of prevalent and incident deformities. AB - Several algorithms are currently in use for evaluating vertebral deformities from plain lateral radiographs of the lumbar and thoracic spine. However, the effects of measurement imprecision as well as uncertainties over image magnification on the correct identification of prevalent and incident vertebral deformities with these algorithms has been little studied. In a pilot study for the European Prospective Osteoporosis Study (EPOS), plain radiographs were submitted to a single central evaluating centre for measurement of vertebral height from T4 to L4. The thoracic and lumbar spines were imaged on separate films, and we have assessed the precision of measurement of vertebral heights and height ratios. The standard deviation of the differences between films of each of three height measurements ranged from 1.1 to 1.2 mm. A two-stage strategy for identifying incident deformities was devised. This required that the vertebra be a prevalent deformity at the time of the second radiograph and also that at least one of the vertebral ratios should have changed significantly since the first radiograph. The second stage removed all but two of the 18 vertebrae flagged positive in the first stage but not considered to be certain incident fractures by clinical reading of the radiographs. PMID- 10673948 TI - Comparison of adult and paediatric spine and whole body software for the Lunar dual energy X-ray absorptiometer. AB - Simple phantoms were devised to compare the performance of adult (software 3.64) and paediatric (software 3.8 g) spine and whole body software developed for the Lunar dual energy X-ray absorptiometer. Rectangular slabs of aluminium with high (1.18 g cm-2) and low (0.57 g cm-2) density were used to represent bone mineral. For spine measurements, the phantoms were scanned in water at depths of 5-20 cm. For whole body measurements, the phantoms were scanned with known amounts of oil and water to represent fat and lean tissue. This simulated tissue depths of 5.5 19.7 cm and body composition ranging from 14-29% fat. There were systematic differences in spine and whole body bone mineral content (BMC), bone area (BA) and bone mineral density (BMD) measurements and also between adult and paediatric software versions. The magnitude and direction of these differences were dependent on BMD of the phantom and tissue depth. Similar systematic differences were observed in vivo when volunteers were scanned using adult and paediatric software. Paediatric software enabled measurements to be made at low tissue depths. The weights of fat, lean and total soft tissue measured by the adult and paediatric whole body software were similar to the values calculated from the known composition of the phantom. Precision estimates for all softwares were excellent. In conclusion, paediatric software should improve bone mineral measurements of children but the discrepancies between adult and paediatric softwares may cause problems in longitudinal studies of skeletal growth and when compiling reference data from infancy through to adulthood. PMID- 10673949 TI - A phantom-based evaluation of an exposure equalization technique in mammography. AB - An anatomical filter based exposure equalization technique in mammography is evaluated quantitatively using a phantom. The evaluation is carried out by a comparative observer performance study, comparing the equalization technique with a conventional one based on visualization of low contrast, 6 mm circular details and high contrast, 0.5 mm and 0.25 mm small size details. These details are situated at the phantom edge, simulating the breast periphery. Visualization of these details is studied with respect to the parameters of tube voltage, optical density, detail location and phantom thickness. Phantom images are interpreted independently by three observers using a four-point grading scale. Use of the Wilcoxon signed ranks test for paired data shows statistically highly significant improvement (p < 0.0001) in the visualization of details for the equalization technique for all values of the parameters studied. The improvement is independent of tube voltage but dependent on optical density, detail location and phantom thickness. Optimal performance is obtained for detail location closer to the outer border of the simulated breast periphery and/or further away from the film, as well as for a greater phantom thickness simulating both thick and dense breast. PMID- 10673950 TI - Reducing cardiac dose in post-operative irradiation of breast cancer patients: the relative importance of patient positioning and CT scan planning. AB - Left-sided post-operative radiotherapy fields for the treatment of breast cancer inevitably encompass the heart within the treatment volume, resulting in late mortality which may negate the cause-specific survival advantage of the therapy. The effect of positioning was studied in 11 patients with left-sided tumours and five with right-sided tumours receiving routine post-operative radiotherapy to the breast or chest wall as part of primary therapy for breast cancer. Using the same arrangement of glancing fields for each patient treatment position, the optimum patient positioning resulted in a reduction in cardiac dose compared to our standard patient treatment position. On the left side the reduction in mean cardiac dose was 60% (p < 0.001) and the reduction in maximum dose was 32% (p < 0.001); on the right it was 17% and 31%, respectively. The volume of cardiac tissue irradiated was also reduced for all patients. Using this optimum treatment position, cardiac dose was investigated in a further 10 patients with left-sided tumours and our standard glancing field set-up was compared with 3-dimensional planning. A further reduction of 12% in the mean cardiac dose was achieved. 5 of 10 patients had a further small reduction of 4.6% in the maximum dose and one patient had a further reduction in maximum dose of 58%. In conclusion, sophisticated radiotherapy planning can reduce cardiac doses, but optimum patient positioning is of greater importance. The general application of such relatively simple measures could have a significant positive effect on overall survival from breast cancer. PMID- 10673951 TI - Small bowel MRI using water as a contrast medium. AB - Magnetic resonance imaging (MRI) of the small bowel has been limited by lack of an adequate luminal contrast medium and problems with image artefacts. In this study we investigate the feasibility of imaging the luminal small bowel using rapid heavily T2w techniques, similar to those used for MR cholangiopancreatography, combined with oral water loading. Eight volunteers were examined after drinking 1-21 of water using serial, multisection, half-Fourier single shot rapid acquisition with relaxation enhancement (RARE) acquisitions. The examinations were continued until the terminal ileum was reached or the water reabsorbed. The results were subjectively assessed for visibility of the small bowel. In all subjects the duodenum, jejunum and ileum were well demonstrated with valvulae conniventes clearly visible. The water column reached the terminal ileum and the caecum in six of the eight subjects but in the remaining two water remained in the small bowel and was ultimately reabsorbed. These preliminary results suggest that with further refinement such an approach may be practical for clinical magnetic resonance imaging of the small bowel. PMID- 10673952 TI - High speed injection of radiographic contrast media induces severe particulate contamination. AB - In a study of exogenous particulate contamination during angiography, the effect of injection speed of four kinds of radiographic contrast media (RCM) was investigated. The particle count (> or = 10 microns) in all RCM increased in a speed-dependent manner and the increase was especially dramatic at 3 ml s-1. The extent of increase in particulate matter was higher for ioxaglate than for the other three RCM. As particulate matter is unwanted and unnecessary, to prevent the harmful effects in patients much attention should be paid to various factors generating particulate matters, such as characteristics of RCM and plastic syringe. PMID- 10673953 TI - The performance of a fluoroscopic electronic portal imaging device modified for portability. AB - Advances in external beam therapy technology have made routine, efficient conformal therapy a reality. With it comes the increasing need for online treatment verification, which is only achievable at present through the use of electronic portal imaging devices (EPIDs). For a large radiotherapy centre, the provision of one EPID per treatment machine proves extremely expensive. This paper details modifications to the design of a commercial fluoroscopic EPID (the SRI-100) to produce a portable system, capable of providing quick, high quality imaging on more than one treatment machine. We describe the necessary hardware and software changes made to the system, as well as the variety of mechanical and quality control checks performed for testing the stability and quality of the imaging. The modified system has been found to be both electronically and mechanically robust, with associated image quality, scaling, distortion and movement similar to other EPIDs in the department. Although the modification was designed specifically to allow for the acquisition of images from multiple treatment machines, it may also enable the operation of the EPID for other uses such as total body irradiation (TBI) treatment verification and a further range of quality control procedures on the linear accelerator itself. PMID- 10673954 TI - Magnetic resonance imaging of actinomycosis presenting as pelvic malignancy. AB - Pelvic actinomycosis is associated with long-standing use of an intrauterine contraceptive device and may present with clinical signs and symptoms of pelvic malignancy. Diagnostic imaging can confirm the presence of a pelvic mass and tissue infiltration but findings are often non-specific. We present a case of pelvic actinomycosis with tubo-ovarian abscess in which magnetic resonance imaging demonstrated lower signal intensity tissue on T2 weighted sequences than would be typical for pelvic malignancy or infection and was useful in confirming regression of pelvic disease in response to antibiotic therapy. PMID- 10673955 TI - Plasma cell neoplasms in the young. AB - Multiple myeloma, a disease of the elderly, is extremely rare in those below 30 years of age. Two patients with multiple myeloma diagnosed at 20 years and 18 years are described. Both presented with extradural cord compression, lytic bone lesions and bone marrow plasmacytosis. One patient received combination chemotherapy and radiotherapy and survived for 14 years. A literature review is presented. PMID- 10673956 TI - A case of rectoprostatic fistula due to prostatic abscess visualized by barium enema. AB - We report a rare case of rectoprostatic fistula due to spontaneous rupture of a prostatic abscess in a patient with diabetes mellitus. Barium enema clearly showed rectoprostatic fistula. Barium enema and colonoscopy were very useful in demonstrating the rectoprostatic fistula. PMID- 10673957 TI - Hepatic artery aneurysm. AB - Hepatic artery aneurysms (HAAs) are rare. A review of the English language literature from 1985 to 1995 for reports of visceral artery aneurysms showed HAA to be the most frequently reported visceral aneurysm during that decade. This increase in incidence relates to the increasing use of percutaneous diagnostic and therapeutic procedures. A second factor is the increased use of diagnostic CT scanning after blunt liver trauma. The purpose of this pictorial review is to illustrate the imaging presentation and radiological management of HAAs. PMID- 10673958 TI - Too much fat in the wrong places. PMID- 10673959 TI - Contrast-enhanced CT in acute pancreatitis. PMID- 10673960 TI - Psychosocial oncology: the state of the art at the end of the millennium. PMID- 10673961 TI - Psycho-oncology: where have we been? Where are we going? AB - This article reviews the development of the subspeciality of psycho-oncology and its contributions to patient care, encouraging more attention to and research into the care of the total patient: the physical, psychological, social and spiritual aspects of care. The result is enhanced quality of life as the patient is studied in the domains of living that are important, extending across the continuum of care from diagnosis to palliative care. In addition, cancer prevention and early detection depends largely on changing attitudes and behaviours that put people at greater risk. This is an important area of research for psycho-oncologists. In the past two decades, research has contributed to our understanding of the psychological responses that accompany a cancer diagnosis. Oncologists better recognise psychological distress and psychiatric disorders such as anxiety, depression and delirium (in hospitalised patients) as frequent comorbid disorders. The development of valid assessment tools for the patients' self-report has been important. Increasingly, outcome measures in controlled trials of new therapies include quality of life, and no longer look at survival alone. The future will continue to bring new challenges to psycho-oncology as patients face new challenges in treatment. A major aim of the next century will be to bring this integrated approach to all patients in an affordable manner. PMID- 10673962 TI - Psychosocial interventions for patients with cancer: what works and what doesn't. AB - With the successes that have been achieved in cancer care leading to patients surviving longer, the need for a variety of psychosocial intervention models is posing a new challenge to the field. This article reviews the general categories of interventions used most commonly: (1) education; (2) coping; (3) emotional support; and (4) psychotherapy. It provides a theoretical model for designing psychosocial interventions, and provides guidelines for assessing what works and what doesn't. PMID- 10673963 TI - What has been learned from measuring health-related quality of life in clinical oncology. AB - The measurement of health-related quality of life (HRQL) in oncology clinical trials has come of age. Most cooperative clinical trials groups as well as individual institutions have either been measuring, or are starting to measure, HRQL. Over the past decade, much has been learned about how to incorporate HRQL components into multicentre, randomised controlled (phase III) trials and how to collect the data with reasonably low levels of missing information. A selective review, focused primarily on phase III studies, shows that HRQL data are useful for deciding which treatment is preferable when survival rates are similar and for determining whether changes in HRQL, as compared with baseline levels, are related to a treatment or intervention. HRQL information is improving our knowledge of the effects of diseases and their treatments on the patient's ability to function and sense of well-being, and HRQL status is proving to be a more accurate predictor of survival than is performance status. Much more remains to be done, but it is apparent that the inclusion of HRQL in clinical trials has been informative and useful. The increasing frequency of HRQL assessment in clinical trials is evidence of the emergence of a patient-centred philosophy in clinical medicine which, in time, will modify the disease-oriented paradigm under which medical professionals have functioned for the past century. PMID- 10673964 TI - Quality of life instruments in oncology. AB - The objective of this article is to aid clinicians in understanding the current state of the development and application of quality of life (QOL) instruments as outcome measures in cancer clinical research and practice. As a result of the achievements of the past two decades, the concept of QOL has been defined and many reliable and valid measurement tools have been developed. The two main approaches to QOL assessment, psychometric-based and utility-based, are discussed together with a brief description of the strategies for meaningful interpretation of QOL profiles. QOL measures in oncology have the potential to be used to study populations in randomised clinical trials, to aid patient-clinician interactions in routine practice and to support policy decision making and economic evaluation of healthcare provision. PMID- 10673965 TI - Improving the quality and quantity of life among patients with cancer: a review of the effectiveness of group psychotherapy. AB - Cancer patients suffer from a number of psychosocial problems related to the progression of their disease as well as standard medical interventions. Fortunately, there is empirical evidence suggesting that group psychotherapy is effective at ameliorating psychological distress and in some cases improving survival. For this literature review we examined the psychological morbidity, particularly anxiety and depression, among cancer patients. Further, we conducted a critical examination of the current evidence regarding the effectiveness of group psychotherapy for improving the quality as well as the quantity of life in cancer patients. Finally, we explored the specific components of effective group psychotherapy, which has been associated with enhanced survival. We conclude that there is compelling evidence indicating that group psychotherapy improves the quality of life of cancer patients. Furthermore, there is a growing body of evidence suggesting that group psychotherapy improves survival of cancer patients. PMID- 10673966 TI - Cancer information: a cost-effective intervention. AB - There is a considerable knowledge base about the information needs of patients with cancer (and their relatives and friends). Those needs will vary according to the disease, the stage of disease, the patient and his or her age, social class and culture. Lack of information may lead to increased anxiety and distress, may impact negatively on the patient's satisfaction and may influence a patient's treatment choices. Other articles in this special edition deal with psychosocial interventions and complementary therapies for cancer patients and explore their efficacy. The reality is that these are unlikely to be made available to all cancer patients for reasons of cost and practicability. Information, however, is a relatively cheap intervention that could--and should--be part of standard care. This article explores some of the research about the provision of information for cancer patients. PMID- 10673967 TI - Effective communication skills are the key to good cancer care. AB - Communication within oncology is a core clinical skill but one in which few oncologists or specialist cancer nurses have received much formal training. Inadequate communication may cause much distress for patients and their families, who often want considerably more information than is usually provided. Many patients leave consultations unsure about the diagnosis and prognosis, confused about the meaning of--and need for--further diagnostic tests, unclear about the management plan and uncertain about the true therapeutic intent of treatment. Additionally, communication difficulties may impede the recruitment of patients to clinical trials, delaying the introduction of efficacious new treatments into clinics. Lack of effective communication between specialists and departments can also cause confusion and a loss of confidence amongst the team. Oncologists themselves acknowledge that insufficient training in communication and management skills is a major factor contributing to their own stress, lack of job satisfaction and emotional burnout. Consequently, over the past few years there have been several initiatives aimed at improving basic communication skills training for healthcare professionals in the cancer field. In this paper, some of the issues that influence communication within an oncology setting, and ultimately affect patient care, are discussed. PMID- 10673968 TI - The impact of patient treatment preferences on the interpretation of randomised controlled trials. AB - Reliable information about aggregate main treatment effects in cancer research comes from randomised controlled trials (RCTs). The possibility of important interactions, such as between treatment preferences and their effects, is necessarily subordinated in the quest for evidence about main treatment effects. If patient preferences can influence the effectiveness of treatments, for which there is some indirect evidence, then those estimates of the treatment's main organic effects from unblind RCTs might be wrong. RCTs clearly disallow patient choice and it is, therefore, important to know the extent of any preference effects in order to interpret the RCT evidence. It may be important to know whether they exist, and where and by how much they affect outcome. It is argued that measuring these effects reliably is methodologically difficult, and will require massive trials each directed at measuring one particular preference effect. Such effects have a slightly fanciful image, particularly in cancer treatment, and may be transient. Given the current uncertainties about their true nature and plausible biological mechanisms, the accumulated evidence is unlikely to provide sufficient justification for investing in such trials, given other current priorities. PMID- 10673969 TI - Psychoneuroimmunology and cancer: fact or fiction? AB - There is substantial evidence from both healthy populations as well as individuals with cancer linking psychological stress with immune downregulation. This discussion highlights natural killer (NK) cells, because of the role that they may play in malignant disease. In addition, distress or depression is also associated with two important processes for carcinogenesis: poorer repair of damaged DNA, and alterations in apoptosis. Conversely, the possibility that psychological interventions may enhance immune function and survival among cancer patients clearly merits further exploration, as does the evidence suggesting that social support may be a key psychological mediator. These studies and others suggest that psychological or behavioural factors may influence the incidence or progression of cancer through psychosocial influences on immune function and other physiological pathways. PMID- 10673970 TI - How useful are unconventional cancer treatments? AB - Unconventional cancer treatments are used frequently. Therefore, oncologists need to know about them. This article gives an overview of current knowledge on the most prevalent complementary or alternative cancer therapies. A distinction is made between alleged cures, preventive and adjunctive measures. Shark cartilage, mistletoe, thymus therapy, essiac, hydrazine sulphate, 714-X, dietary regimens, green tea and Panax ginseng are all covered specifically. None of these treatments offer reasonable hope for a cure. Some strategies are promising in terms of cancer prevention. The true potential of unconventional therapies might lie in adjunctive and palliative care. It is concluded that good evidence in this area is scarce. Vis-a-vis the high prevalence of unconventional cancer treatments, rigorous investigations are mandatory, not least for increasing the safety of future patients. PMID- 10673971 TI - Testing complementary and alternative therapies within a research protocol. AB - In patients with cancer, the demand for complementary and alternative medicine (CAM) is considerable. Unfortunately, however, for many of these interventions there is a lack of evidence for efficacy, effectiveness and safety in patients with cancer. This review focuses on the prospective, randomised, controlled trial (RCT) as a tool for evaluating CAM. Although a number of difficulties and limitations are acknowledged, the RCT will continue to be the gold standard for evaluating the efficacy, effectiveness and safety of CAM. Developments in clinical trial methodology and in psychosocial oncology have made it more appropriate and feasible to evaluate CAM using RCT methodology. Two different kinds of RCTs are now accepted as valid, namely explanatory and pragmatic trials. The latter does not necessarily require that the patient or the therapist is 'blind' to the treatment being given. Furthermore, pragmatic trials can be designed to take patient preferences into account. A number of practical issues are discussed, including the choice of comparator or control interventions, ways of assessing the effects of individual differences, minimising therapist variability, the problem of finding acceptable inclusion-exclusion criteria and the assessment of treatment outcome. A number of randomised, controlled trials have demonstrated the efficacy, effectiveness and safety of various complementary and alternative interventions (the Cochrane Data Base has now established a CAM field). The publication of positive results from randomised trials of complementary interventions that have not yet been studied using this methodology would do a great deal to alleviate the scepticism of conventional practitioners towards these types of CAM and would facilitate further the integration of complementary and conventional interventions. PMID- 10673972 TI - Improving communication with cancer patients. AB - If doctors and nurses involved in cancer care are to help patients and their families achieve an optimal level of quality of life and psychological adjustment they must be able to carry out key communication tasks successfully. Yet, objective scrutiny of their consultations confirms that deficiencies in their ability to conduct these tasks remain. The reasons for this are discussed before important innovations in training and their impact are described. PMID- 10673973 TI - EORTC scientific strategy meeting 25-26 March 1999. PMID- 10673974 TI - Efficacy and safety of docetaxel (Taxotere) in heavily pretreated advanced breast cancer patients: the French compassionate use programme experience. AB - The aim of this investigation was to assess retrospectively docetaxel safety and efficacy in advanced breast cancer patients in a French compassionate use programme. Patients had received > 1 prior chemotherapy regimen for advanced disease, were either anthracycline-resistant (that is progressed within 6 months after anthracycline-based chemotherapy) or had received the maximum cumulative dose. The recommended docetaxel dose was 100 mg/m2/cycle (75 mg/m2 in case of liver function impairment: transaminases > 1.5 x upper limit of normal (ULN), alkaline phosphatases > 3 x ULN). Between August 1993 and December 1995, 889 patients were treated in 67 French centres, of whom 870 were evaluable for safety and 825 were evaluable for patient and treatment characteristics and efficacy. 20.5% (of the 825 patients evaluable for baseline characteristics) had poor performance status (PS > or = 2), 49.3% liver metastasis and 9.6% biological liver dysfunction. 98.4% had been previously treated by anthracyclines, 50.8% had resistant disease and 37.1% had received > 2 prior palliative chemotherapy lines. The most frequent severe toxicity, febrile neutropenia (reported in 223/870 (25.6%) patients evaluable for safety), caused 10 deaths, 6 of these being patients with severe liver impairment before inclusion. Fluid retention syndrome and other common non-haematological toxicities were well tolerated. 3.1% (28/889) of all patients and 11.4% of those with liver dysfunction, died from treatment related causes. The overall response rate in 825 assessable patients was 22.9% (95% confidence interval (CI): 20.2-26.2%). Median time to treatment failure was 4 months (95% CI: 3.6-4.3) and median survival was 9.8 months (95% CI: 8.8-10.7). This report on the largest series of unselected advanced breast cancer patients treated with docetaxel, supports previous phase II studies, confirming docetaxel's utility in patients relapsing after failing anthracycline-containing palliative chemotherapy. PMID- 10673975 TI - Centre effect on treatment outcome for patients with untreated acute myelogenous leukaemia? An analysis of the AML 8A Study of the Leukemia Cooperative Group of the EORTC and GIMEMA. European Organization for Research and Treatment of Cancer (EORTC) Leukemia Cooperative Group and the Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto (GIMEMA). AB - In the AML 8A study patients were treated with remission-induction therapy followed by one consolidation course. Patients in complete remission (CR) were randomised between autologous bone marrow transplantation (ABMT) and a second intensive consolidation course, except for those with a histocompatible sibling donor, who received allogeneic bone marrow transplantation (alloBMT). This analysis was performed to determine whether centres which only performed induction and consolidation therapy, achieved similar results as centres who also performed transplantation. 542/676 (80%) from transplantation centres and 150/194 (77%) from referring centres achieved CR, with an early death rate of 5% and 11%, respectively (P = 0.01). 66% of patients with a donor from transplantation centres received alloBMT in first CR compared with 57% from referring centres (P = 0.2). Transplantation centres randomised 64% of patients without a donor, referring centres 47% (P = 0.04). The full protocol treatment was completed by 275/542 (51%) and 61/150 (41%) patients, respectively (P = 0.04). The overall survival rate at 6 years from diagnosis was 34% and 36%, respectively (P = 0.9). In conclusion, the type of centre did not appear to have an influence on overall survival. The feasibility of the study was acceptable for both types of centres. The referring centres applied more selection for transplantation. Despite a more intensive second-line treatment at transplantation centres, the overall outcome remained similar to that of referring centres. PMID- 10673976 TI - The effects of the participation of patients with cancer in teaching communication skills to medical undergraduates: a randomised study with follow-up after 2 years. AB - The importance of good doctor-patient communication is widely recognised. The aims of this study were to evaluate the immediate effects of the participation of patients with cancer on the attitudes and skills of undergraduate medical students receiving an interview skills training programme, and to assess the effects of the participation of patients with cancer on the attitudes and interview performance of students 2 years later. It was hypothesised that the participation of cancer patients would have specific beneficial effects on attitudes and interview performance. Before participating in a 6-session interview methods course in third year, students were randomised to be taught with patients who had cancer (experimental group) or with patients with other diagnoses (control group). Before and after participating in the course, 233 students (94% response rate) completed an Attitudes Questionnaire. When they reached their fifth year, 54 students again completed the Attitudes Questionnaire and, in addition, made a video recording of an interview with a patient who had gynaecological cancer. These recordings were rated independently by two researchers using the Interview Rating Instrument. Immediately after the course, a number of differences were found between the two groups. For example, students in the experimental group were more likely to consider the ability to listen an extremely important characteristic of hospital doctors and to consider more strongly that trust is an essential part of the doctor-patient relationship. 2 years after the course, the ability of hospital doctors to communicate with patients, and the need for clinical decisions to reflect patients' wishes, were considered to be more important by students in the experimental group, although even 96% of controls felt both these issues were very or extremely important. As hypothesised, the experimental group had better ratings in terms of responding empathically, showing regard and concern for the patient, and assessing the impact of the symptoms on the patient's life. The participation of patients with cancer has beneficial and enduring effects on the attitudes and interview performance of medical undergraduates. Medical schools should consider how best patients with cancer can make an important contribution to communication skills training. PMID- 10673977 TI - Ifosfamide, vinorelbine and gemcitabine in advanced non-small cell lung cancer. A phase I study. AB - The objective of this phase I study was to identify the maximum tolerated dose (MTD) and toxicity of a three drug, platinum-free regimen, including gemcitabine, ifosfamide and vinorelbine, in the treatment of patients with advanced non-small cell lung cancer (NSCLC). 33 chemotherapy-naive patients with histologically confirmed, unresectable NSCLC, received fixed doses of ifosfamide (1500 mg/m2 days 1-3 with mesna) and vinorelbine (25 mg/m2 days 3 and 8). The gemcitabine dose was escalated from 500 to 1200 mg/m2 on days 3 and 8 every third week. The escalation was stopped at dose level 4 (gemcitabine 1200 mg/m2) since all 3 patients of this cohort showed dose-limiting thrombocytopenia and/or neutropenia at treatment cycle 1. The dose recommended for phase II trials is: gemcitabine 1000 mg/m2 and vinorelbine 25 mg/m2 given on days 3 and 8 plus ifosfamide 1500 mg/m2 on days 1-3. An encouraging response rate of 50% (95% confidence interval (CI): 32-68%) was observed in 32 patients evaluated. Our results show that ifosfamide, vinorelbine and gemcitabine can be safely administered as outpatient chemotherapy for NSCLC. Myelosuppression is the dose-limiting toxicity (DLT) of this regimen with no major subjective side-effects observed. PMID- 10673979 TI - Paediatric Hodgkin's disease. PMID- 10673978 TI - Fas expression in non-small cell lung cancer: its prognostic effect in completely resected stage III patients. AB - The aim of this study was to examine Fas expression in non-small cell lung cancer (NSCLC) and examine its correlation with clinicopathological features and prognosis. Fas expression was determined by an immunohistochemical analysis using the labelled streptavidin-biotin method from 220 paraffin specimens of completely resected primary stage I-III NSCLC. 80 (36%) of 220 cases were positive for Fas immunostaining. These 80 cases included 44 adenocarcinomas (33%) and 30 squamous cell carcinomas (40%). 33 stage I (33%) 13 (43%) stage II and 34 (37%) stage III tumours were Fas positive. No statistically significant differences were observed regarding the Fas status with respect to age, sex, histological type, or stage of disease. There was no significant difference in survival between early stage (stages I-II) disease patients with positive Fas expression and those with a negative expression (P = 0.719). However, for patients with completely resected stage III tumours, the patients with positive Fas staining were found to survive for a longer period than those with negative staining (P = 0.026). PMID- 10673980 TI - Cancer mortality in Europe, 1990-1994, and an overview of trends from 1955 to 1994. AB - Mortality data, abstracted from the WHO database, are presented in tabular form for 26 cancer sites or groups of sites, plus total cancer mortality, in 35 European countries during the period 1990-1994. Trends in mortality are also given in graphical form for 24 major countries over the period 1955-1994. In most western European countries total cancer mortality was--for the first time- moderately downwards in the early 1990s. Such favourable trends included some decline in lung cancer mortality for males, the persistent decline in stomach cancer for both sexes, and of cervical cancer for women, as well as some decline in breast and colorectal cancers, plus other neoplasms (testis, lymphoid neoplasms), whose treatment has further improved over the last few years. However, cancer mortality was still upwards in a few southern and eastern European countries, including Hungary and Poland, where total cancer mortality rates in middle-aged males are now the highest ever registered in Europe. The favourable trends in western Europe over the recent years are similar to those observed in the U.S.A. PMID- 10673981 TI - Induction of apoptosis by apigenin and related flavonoids through cytochrome c release and activation of caspase-9 and caspase-3 in leukaemia HL-60 cells. AB - The aim of this study was to investigate the mechanism of flavonoid-induced apoptosis in HL-60 leukaemic cells. Thus, the effect of structurally related flavonoids on cell viability, DNA fragmentation and caspase activity was assessed. Loss of membrane potential and reactive oxygen species generation were also monitored by flow cytometry. The structurally related flavonoids, such as apigenin, quercetin, myricetin, and kaempferol were able to induce apoptosis in human leukaemia HL-60 cells. Treatment with flavonoids (60 microM) caused a rapid induction of caspase-3 activity and stimulated proteolytic cleavage of poly-(ADP ribose) polymerase (PARP). Furthermore, these flavonoids induced loss of mitochondrial transmembrane potential, elevation of reactive oxygen species (ROS) production, release of mitochondrial cytochrome c into the cytosol, and subsequent induction of procaspase-9 processing. The potency of these flavonoids on these features of apoptosis were in the order of: apigenin > quercetin > myricetin > kaempferol in HL-60 cells treated with 60 microM flavonoids. These results suggest that flavonoid-induced apoptosis is stimulated by the release of cytochrome c to the cytosol, by procaspase-9 processing, and through a caspase-3 dependent mechanism. The induction of apoptosis by flavonoids may be attributed to their cancer chemopreventive activity. Furthermore, the potency of flavonoids for inducing apoptosis may be dependent on the numbers of hydroxyl groups in the 2-phenyl group and on the absence of the 3-hydroxyl group. This provides new information on the structure-activity relationship of flavonoids. PMID- 10673982 TI - Low-doses of ionising radiation induce melanoma metastases and trigger the immune system--adrenal axis feedback loop. AB - Low-doses of ionising radiation are frequently implicated in triggering and/or accelerating the growth of skin and other malignancies. It seemed probable that the radiation at similar dose levels might initiate metastasis from already existing tumours. Highly pigmented human melanoma xenograft that had lost its ability for a spontaneous metastasising and grown subcutaneously in athymic mice was exposed to very low and well-defined doses of ionising radiation to determine whether low linear energy transfer radiation can restore metastatic potential of the tumour. To ensure that all effects derived from radiation-activated neoplastic cells only, I was delivered selectively to the cutaneous melanoma instead of using the external beam. The direct response of these tumours to radiation was monitored by determining the growth rate of the lesions. Histopathological methods were employed to detect metastases. The lowest radiation dose of approximately 6 cGy deposited in the tumours initiated metastatic spread in all animals. Gradual increase of the radiation doses diminished both the frequency of the appearance of metastases and their distance from the primary lesions. There were no metastases from non-irradiated melanomas. The highest dose used (60 cGy) did not affect significantly the growth of cutaneous (primary) tumours, but lower doses that enhanced inflammatory infiltration of the lesions reduced tumour growth. Such radiation-stimulated immune responses were accompanied by increased pigmentation in cutaneous lesions and activation of the adrenal cortex indicating that the immune system-adrenal axis feedback loop had been triggered. The results demonstrate that very low doses of ionising radiation induce melanoma metastases. The phenomenon is accompanied by the stimulation of the immune system-adrenal axis feedback loop that regulates eicosanoid synthesis, thereby suggesting an involvement of these molecules in the process. Radiation doses approaching the therapeutic level do not initiate melanoma dissemination. PMID- 10673983 TI - Effect of gamma-linolenic acid on cellular uptake of structurally related anthracyclines in human drug sensitive and multidrug resistant bladder and breast cancer cell lines. AB - This study investigated the effect on drug uptake in multidrug resistant cells by the incorporation of the essential fatty acid gamma-linolenic acid (GLA). The cell lines used were the MCF-7/R resistant human breast cancer and MGH-U1/R bladder cancer. Uptake of drug (doxorubicin, epirubicin, mitoxantrone and idarubicin) after the incorporation of GLA was investigated quantitatively by flow cytometry and qualitatively by confocal microscopy. There was no observable overall increase in drug uptake due to GLA incorporation into the cells as shown by flow cytometry. However, an increase in uptake of the chemotherapeutic agent idarubicin was observed in GLA-treated resistant cells compared with untreated cells using the confocal microscope. This overall increase in cellular drug uptake was not accompanied by a change in cellular drug distribution. Only one drug, mitoxantrone, displayed a change in intracellular drug distribution due to GLA incorporation into MCF-7/R cells. This suggests that essential fatty acid incorporation into the cellular membranes of some resistant cells may cause a shift in the intracellular distribution of certain chemotherapeutic drugs. PMID- 10673984 TI - Inhibition of P-glycoprotein activity and reversal of multidrug resistance in vitro by rosemary extract. AB - The transmembrane transport pump P-glycoprotein (Pgp) causes the efflux of chemotherapeutic agents from cells and is believed to be an important mechanism in multidrug resistance (MDR) in mammary tumours. In the present study we demonstrate that an extract of the common dietary herb rosemary (Rosemarinus officinalis Labiatae), increases the intracellular accumulation of commonly used chemotherapeutic agents, including doxorubicin (DOX) and vinblastine (VIN), in drug-resistant MCF-7 human breast cancer cells which express Pgp. Rosemary extract (RE) inhibits the efflux of DOX and VIN, which are known to be substrates of Pgp, but does not affect accumulation or efflux of DOX in wild type MCF-7 cells, which lack Pgp. Treatment of drug-resistant cells with RE increases their sensitivity to DOX, which is consistent with an increased intracellular accumulation of the drug. RE blocks the binding of the VIN analogue azidopine to Pgp. Thus, it appears that RE directly inhibits Pgp activity by inhibiting the binding of drugs to Pgp. PMID- 10673985 TI - Proto-oncogenes and p53 protein expression in normal cervical stratified squamous epithelium and cervical intra-epithelial neoplasia. AB - The aim of this study was to study the protein expression of six proto-oncogenes (epidermal growth factor receptor (EGFR), c-fms, c-myc, c-kit, c-erbB-2 and pan ras) and one tumour suppressor gene (TP53), by immunohistochemical staining of normal cervical stratified squamous epithelium and cervical intra-epithelial neoplasia (CIN). Paraffin sections of 45 normal cervical specimens, 38 CIN grade one (CIN1), 37 CIN2 and 43 CIN3 were studied. An immunohistochemical (IHC) score was derived from the intensity of staining and the percentages of cells stained. In normal cervical specimens, a higher IHC score was found with EGFR and c-fms in superficial (S), intermediate (I) and parabasal (PB) cells compared with basal cells. In contrast, a higher IHC score was found with c-erbB-2 in basal cells in normal cervical specimens. Dysplastic cells in CIN had a higher IHC score with c myc and c-erbB-2 than normal S/I and PB cells. Dysplastic cells had a higher score with EGFR than normal basal cells. However, a higher IHC score with EGFR and c-fms was found in normal S/I cells than dysplastic cells. These findings suggested that EGFR and c-fms were activated in more differentiated normal cells but were less active in less differentiated normal basal cells. However, EGFR was reactivated in dysplastic cells. Meanwhile, c-erbB-2 was activated in less differentiated normal basal cells and dysplastic cells, and was less active in differentiated normal cells. c-myc was activated in dysplastic cells. c-fms was more active in more differentiated normal cells and was not activated in less differentiated or dysplastic cells. c-kit, pan-ras and TP53 were not activated in normal nor dysplastic cervical cells. These results suggest EGFR, c-erbB-2 and c myc may be important proto-oncogenes in CIN and that antibodies or anti-genes targeted against them may alter the progress of CIN to invasive cancer. PMID- 10673986 TI - Comments on: Thoracic radiation therapy before autologous bone marrow transplantation in relapsed or refractory Hodgkin's disease, Tsang, et al. Eur J Cancer 1999, 35, 73-78. PMID- 10673987 TI - Recurrence in the conserved breast: why all this fuss about risk factors? PMID- 10673988 TI - Cancer chemoprevention: progress and promise. AB - Cancer chemoprevention is the use of agents to inhibit, delay or reverse carcinogenesis. The focus of chemoprevention research in the next millennium will include defining the genotypic and phenotypic (functional and histological) changes during carcinogenesis, the cancer risk conferred by these changes, their modulation in preclinical experimentation and randomised clinical trials by chemopreventive drugs, dietary agents and regimens and treatments resulting from early detection. The key elements of this research effort will be basic and translational risk evaluation programmes; chemopreventive and dietary agent drug discovery and development; development of transgenic animal models; required safety and pharmacology studies; well-designed phase I, II and III chemoprevention studies; and much expanded early detection programmes. The large number of chemoprevention research programmes now ongoing ensures that the promise of chemoprevention will continue to be realised in the next decade. PMID- 10673989 TI - Is postoperative irradiation after radical prostatectomy necessary? PMID- 10673990 TI - A comprehensive geriatric assessment (CGA) is necessary for the study and the management of cancer in the elderly. PMID- 10673991 TI - Measurement of clinical and subclinical tumour response using [18F] fluorodeoxyglucose and positron emission tomography: review and 1999 EORTC recommendations. European Organization for Research and Treatment of Cancer (EORTC) PET Study Group. AB - [18F]-fluorodeoxyglucose ([18F]-FDG) uptake is enhanced in most malignant tumours which in turn can be measured using positron emission tomography (PET). A number of small clinical trials have indicated that quantification of the change in tumour [18F]-FDG uptake may provide an early, sensitive, pharmacodynamic marker of the tumoricidal effect of anticancer drugs. This may allow for the introduction of subclinical response for anticancer drug evaluation in early clinical trials and improvements in patient management. For comparison of results from smaller clinical trials and larger-scale multicentre trials a consensus is desirable for: (i) common measurement criteria; and (ii) reporting of alterations in [18F]-FDG uptake with treatment. This paper summarises the current status of the technique and recommendations on the measurement of [18F]-FDG uptake for tumour response monitoring from a consensus meeting of the European Organization for Research and Treatment of Cancer (EORTC) PET study group held in Brussels in February 1998 and confirmed at a subsequent meeting in March 1999. PMID- 10673992 TI - Psychological factors can predict the response to primary chemotherapy in patients with locally advanced breast cancer. AB - This study evaluated the possible value of psychological variables in predicting clinical and pathological response to primary chemotherapy. 96 women with newly diagnosed large, or locally advanced, breast cancer (T2 > 4 cm, T3, T4, N2 and M0) participated in a prospective, randomised trial to evaluate the effects of relaxation training with guided imagery and L-arginine on response to primary chemotherapy. Before the first of six cycles of primary chemotherapy, women were assessed using the Hospital Anxiety and Depression Scale (HADS) and the Eysenck Personality Questionnaire (EPQ). The primary outcomes were clinical response (evaluated using standard International Union Against Cancer (UICC) criteria) and pathological response (graded by means of a previously published 5-point scale) following primary chemotherapy. Stepwise linear regressions were used to estimate the predictive value of age, menopausal status, clinical nodal status, tumour size at diagnosis, oestrogen receptor status, dietary supplementation (L-arginine versus placebo), personality (EPQ-L scores), mood (HADS scores) and a psychological intervention. HADS depression score was a significant independent predictor of pathological response to chemotherapy. HADS anxiety score was a significant independent predictor of clinical response. Because the original tumour size before chemotherapy (also a significant predictor of clinical and pathological responses) was taken into account in the analyses, the results cannot be explained in terms of psychobiological factors related to tumour size. This study supports the importance of psychological factors as independent predictors of response to primary chemotherapy in patients with breast cancer. If they can be replicated, these findings have major implications for the management of women with breast cancer. Psychological factors need to be assessed and evaluated in future trials of chemotherapy. PMID- 10673993 TI - The costs of managing patients with advanced colorectal cancer in 10 different European centres. AB - With the aim of estimating and comparing the direct hospital costs of managing patients with advanced colorectal cancer in various countries, data on resource utilisation and unit prices were collected. Data on the consumption of medical resources were collected by a retrospective examination of the hospital charts for 20 patients in each of 10 centres in five European countries. To make cost comparisons meaningful, a complete and consistent set of unit prices for all the medical resources used in each of the countries would be required, but this could not be achieved. As an alternative method of comparison, the most complete set of unit prices (from Belgium) was used here to estimate the imputed average total cost of patient management in each centre. By using this approach, a summary index was created, which reflected only differences in resource utilisation. This index showed that there were considerable differences in the amounts of resources used for treating these patients, between, as well as within, countries. Differences of the same order of magnitude were found, when the treatment of subgroups of patients, according to site and stage of disease, were examined. PMID- 10673994 TI - Irinotecan in second-line treatment of metastatic colorectal cancer: improved survival and cost-effect compared with infusional 5-FU. AB - In a recent multicentre, randomised, controlled, open-label study (Rougier and colleagues, Lancet 1998, 352, 1407-1412), irinotecan significantly increased survival without any deterioration in quality of life compared with best estimated infusional 5-fluorouracil (5-FU) therapy in the setting of second-line treatment for metastatic colorectal cancer. The aim of the cost-effectiveness analysis reported here was to compare the economic implications, from a U.K. perspective, of replacing 5-FU therapy [either as a single agent (Lokich regimen, B2) or in combination with folinic acid (de Gramont regimen, B1, or AIO regimen, B3)] with irinotecan as second-line therapy for metastatic colorectal cancer. Resource utilisation data collected prospectively during the study, supplemented by both a questionnaire to investigators and local expert clinical opinion, were used as a basis for estimating cumulative drug dosage, chemotherapy administration and treatment of complications. Drug acquisition costs were derived from the British National Formulary (March 1998), and unit costs for clinical consultation and services were derived from relevant 1996/1997 cost databases. Although cumulative drug acquisition costs per patient were higher with irinotecan than with infusional 5-FU therapy, these were at least partially offset by lower cumulative costs per patient associated with administration of therapy and treatment of complications in the irinotecan arm than in the 5-FU arm. Based on the incremental costs per life year gained (LYG), irinotecan was considered to be cost-effective by commonly accepted criteria compared with either the B1 or B2 regimens. Irinotecan was cost-saving compared with the B3 regimen (that is significant survival gain and a reduction in costs). Thus, not only is there strong evidence for the use of irinotecan as standard second-line therapy in metastatic colorectal cancer,but the results of this prospective economic evaluation have shown that irinotecan also represents good value for money in this clinical setting. PMID- 10673995 TI - A phase II study of high-dose octreotide in patients with unresectable pancreatic carcinoma. AB - This report describes the results of a phase II trial to evaluate the safety, feasibility and response of patients with irresectable, histologically proven, stage II-IV adenocarcinoma of the pancreas receiving high-dose octreotide treatment. Octreotide was self-administered subcutaneously (3 x 2000 micrograms per day) by 49 patients. Therapy was discontinued after progression of the disease. Due to the subseqment diagnosis of bile duct carcinoma and stage I disease, 2 patients were excluded, leaving 47 evaluable patients with measurable disease. The median Karnofsky score was 80%. 3 patients had stage II (6%), 19 stage III (40%), and 25 (53%) stage IV disease. Octreotide treatment resulted in stable disease in 9 patients (19%) for more than 12 weeks. No complete or partial response was observed. The median overall survival was 21.4 weeks and the median progression-free survival 9.0 weeks. Therapy with high-dose octreotide is feasible, well tolerated and might prolong survival. In a placebo-controlled phase III study the effects of octreotide in patients with pancreatic cancer will be confirmed. PMID- 10673996 TI - Regression of AIDS-related Kaposi's sarcoma following antiretroviral therapy with protease inhibitors: biological correlates of clinical outcome. AB - The clinical response of AIDS-related Kaposi's sarcoma (KS) to highly active antiretroviral therapy (HAART), a combination of human immunodeficiency virus type 1 (HIV-1) protease and reverse transcriptase inhibitors, was studied in 11 patients, all but one with progressive KS. CD4+ cell counts, plasma HIV-1 RNA levels, and antibody titres to lytic ORF65 and latency-associated human herpes virus type 8 (HHV-8) proteins were determined in sequential samples. Six complete and three partial clinical responses were achieved in a median time of 6 and 3 months, respectively, and confirmed after a median time of 16 months on HAART. 2 patients showed disease progression. A consistent decrease in HIV-1 RNA levels, paralleled by an increase in CD4+ cell counts, was observed in all patients who showed complete or partial clinical response; HIV-1 RNA levels remained persistently high in the two patients who progressed, despite a change in HAART. HHV-8 antibody titres were generally higher in patients with mucosal/visceral involvement compared with patients with limited disease; a decrease in ORF65 antibody titre was significantly associated with a clinical response. These results indicate that HAART is effective for AIDS-related KS; the clinical response correlates with a decrease in plasma HIV-1 RNA levels, an increase in CD4+ lymphocytes, and a decrease in antibodies to ORF65 HHV-8 protein. PMID- 10673997 TI - Survival from childhood cancer in Yorkshire, U.K.: effect of ethnicity and socio economic status. AB - The effect of ethnicity and socio-economic status on the survival of a population based cohort of 1979 children diagnosed with cancer between 1974 and 1995 was investigated. Ethnicity was assigned by computer algorithms and visual inspection as south Asian (or not) for each child, based on their full name. Socio-economic status was measured using the Carstairs index, based on census areas of case residence at diagnosis. 15 children (0.8%) were lost to follow-up. Log-rank tests showed survival from all cancers did not differ between south Asians and other children and no increased risk was observed for south Asians in any diagnostic category, although numbers were small. Increasing levels of deprivation were associated with significant trends of poorer survival from all cancers, leukaemias and brain tumours. Risk of death was typically higher for children from the most deprived areas although differences were not statistically significant after accounting for other factors including ethnicity. Taking all children with malignant disease together, neither ethnicity nor socio-economic status appear to influence survival after taking other factors into consideration. PMID- 10673998 TI - Frequency and nature of germline Rb-1 gene mutations in a series of patients with sporadic unilateral retinoblastoma. AB - Constitutional Rb-1 gene mutations were studied in a series of 17 families with isolated unilateral retinoblastoma patients. Peripheral blood lymphocytes were analysed by karyotyping, Southern blot hybridisation, and 'exon by exon' sequencing. Mutations were detected in 4 (24%) of the investigated probands. All mutations were identified by sequencing. No alteration was detected by Southern blotting or karyotyping. In one of our cases with a R358 stop codon mutation, retinoblastoma was unilateral at the time of diagnosis, but a tumour of the second eye was diagnosed after 35 months of follow-up. After exclusion of this case, the frequency of constitutional mutations in our series was 19% (3 of 16 cases). Alterations in our cases without involvement of the second eye included G ->A substitution in the promoter region 198 bp upstream of the initiating methionine codon; G-->C transversion in the splice donor site at position +1 leading to exon 6 skipping and a 137 bp in-frame deletion, starting 3 bp from the 5' end of exon 15 to 27 bp from the 3' end of exon 16. All alterations were germline de novo abnormalities. PMID- 10673999 TI - Histological determinants for different types of local recurrence after breast conserving therapy of invasive breast cancer. Dutch Study Group on local Recurrence after Breast Conservation (BORST) AB - The purpose of this study was to determine which histological factors are associated with an increased risk for local recurrence in the breast after breast conserving therapy for early breast cancer (TNM stage I and II) and whether risk patterns vary according to menopausal status and type of local recurrence. Through complete follow-up of the patients of eight regional radiation oncology departments, two cancer institutes and one surgical clinic in The Netherlands, 360 patients were identified with local recurrence in the breast after having received breast-conserving therapy (local tumour excision, axillary dissection and irradiation of the whole breast and a boost to the tumour bed) during the 1980s. For each case, two controls with a follow-up of similar duration without local recurrence were randomly selected. Histological slides of the primary tumour were reviewed. Among premenopausal patients the risk of recurrence for those younger than 35 years was significantly higher than that for premenopausal patients of 45 years or older (relative risk (RR) 2.9; 95% confidence interval (95% CI) 1.3-6.6, P < 0.05). The risk of recurrence at or near the site of the primary tumour was most significantly increased for patients with high grade extensive intraductal component (EIC) adjacent to the primary tumour (RR 4.1; 95% CI 1.7-9.8, P < 0.01). Microscopic margin involvement was an important risk indicator for diffuse recurrence and recurrence in the skin of the breast, especially in the presence of vascular invasion (RR 25; 95% CI 4.0-150, P < 0.001). To prevent local recurrence at or near the site of the primary tumour, local excision with a 1-2 cm margin of healthy tissue and a 15 Gy boost seemed adequate local treatment for patients with well differentiated EIC. In contrast, a wider surgical margin, a higher boost dose or mastectomy should be considered for patients with poorly differentiated EIC. Microscopic margin involvement in the presence of vascular invasion significantly increases the risk of diffuse recurrence or recurrence in the skin. PMID- 10674000 TI - Allergy and other selected diseases and risk of colorectal cancer. AB - It has been reported that allergy and other diseases may be related to colorectal cancer risk. The aim of this study was to perform a systematic analysis using information about medical histories specifically to see if there was any relation between allergies or other medical conditions and colorectal cancer risk. A multicentric case-control study was conducted in six Italian areas between 1992 and 1996 on 1225 incident cases of colon cancer, 728 cases of rectal cancer and 4154 controls comparable with cases according to sex and age group, admitted for acute conditions to the same network of hospitals where cases had been identified. Unconditional logistic regression models including terms for sex, age, study centre, years of education, body mass index, physical activity, smoking, history of colorectal cancer in first-degree relatives and energy intake were used to estimate the odds ratios (OR) of colon and rectal cancer according to history of allergy and other selected diseases. The OR for history of allergy was 0.88 (95% confidence interval, CI, 0.67-1.14) for colon and 0.64 (95% CI, 0.44-0.92) for rectal cancer, and the inverse association was stronger when allergy was diagnosed at age 35 years or more, or less than 10 years before the cancer diagnosis. No clear pattern emerged in strata of age and sex. History of other selected diseases, including hypertension and cholelithiasis, was not related to colon or rectal cancer risk, though there was a moderate increase in the risk of colon cancer (OR = 1.18, 95% CI, 0.66-2.14) in patients with a history of intestinal polyps. This study lends support to the hypothesis that allergic individuals may be at a lower risk of developing colorectal cancer. PMID- 10674001 TI - Chemical hepatocarcinogenesis in transgenic mice overexpressing mature TGF beta-1 in liver. AB - The role of transforming growth factor beta 1 (TGF-beta 1) in carcinogenesis is a controversial issue. Certain results suggest a promoter role of this growth factor whilst in other experimental models TGF-beta 1 seems to inhibit the process of tumorigenesis. In an attempt to resolve this problem, we have performed chemical hepatocarcinogenesis experiments on transgenic mice expressing a high level of active TGF-beta 1 in their liver. Transgenic production of TGF beta 1 did not result in spontaneous tumour formation during our observation period. However, two carcinogens, thioacetamide and N-OH acetylaminofluorene, were more potent in transgenic than in wild-type mice, whereas aflatoxin B1 was equally effective in both groups. Our observations suggest that an increased level of TGF-beta 1 in the liver does not provide protection against the effect of chemical carcinogens. PMID- 10674002 TI - Pharmacokinetic differences between rat tumour and lung tissues following isolated lung perfusion with cisplatin. AB - Isolated lung perfusion has been performed for the treatment of unresectable lung tumours; however, the pharmacokinetics of this procedure remain unclear. This study was conducted to investigate the changes in antitumour drug concentrations in tumour and lung tissues after isolated lung perfusion, using different perfusion times and perfusate drug concentrations. Isolated left lungs were perfused for 20, 40 or 60 min with 25, 50 or 100 micrograms/ml of cisplatin after solitary lung tumour nodules were established in rats, and the total platinum concentrations in the perfused lung and tumour tissues were determined by flameless atomic absorption spectroscopy. The oedema in the perfused lung tissues was evaluated by histological examination and by the wet to dry weight ratios of the lungs. The total platinum concentration increased significantly with perfusion time and increasing perfusate cisplatin concentrations in the lung tissue, but it did not change in the tumour tissue. The wet to dry weight ratios of the lung tissues did not differ significantly among the perfusion groups. Oedema of the perfused lung tissue did not change significantly with the perfusion time or perfusate cisplatin concentration. The results of this study indicate the possibility that different pharmacokinetics exist between tumour and lung tissues following isolated lung perfusion with cisplatin, which could be used as a clinical guide for the selection of appropriate perfusion times and perfusate drug concentrations. PMID- 10674003 TI - Sequence-dependent growth inhibition and DNA damage formation by the irinotecan-5 fluorouracil combination in human colon carcinoma cell lines. AB - We evaluated irinotecan (CPT-11) together with 5-fluorouracil (5-FU) for improved cell growth inhibition with respect to that by either agent alone in the human colon carcinoma cell lines SW620, HT-29 and SNU-C4. Cells were exposed for 24 h to each drug, as well as to various combinations and sequences of low, fixed doses of one drug with higher varying doses of the other, cultured for two more days in drug-free medium and then assessed for growth response with the sulphorhodamine B assay. Multiple drug effect analysis was used to evaluate the data, which were then related to the amount of DNA damage occurring in the cells which was determined by a fluorescence-enhancement assay for DNA unwinding. Cellular responses were also related to thymidylate synthase topoisomerase I and carboxyl esterase activities, which were assessed by a ligand-binding and a 3H release assay; a DNA decatenation assay; and a spectrophotometric method, respectively. IC50 values for 5-FU alone in the SW620, HT29 and SNU-C4 cells were 15.3 +/- 0.8, 8.2 +/- 1.3 and 2.2 +/- 0.7 microM, respectively, and for CPT-11 2.0 +/- 0.9, 2.5 +/- 0.5 and 3.8 +/- 0.3 microM, respectively. The differential responses to 5-FU alone were possibly determined by differences in substrate affinity and conversion rate of thymidylate synthase (K(m) of approximately 7.5, 5.0 and 2.5 microM and V0 of approximately 800, 200 and 2400 microM/h, respectively). The comparable cellular responses to CPT-11 alone might be accounted for by the counterbalancing effects of differences in topoisomerase I (1, 1, and 1.5 arbitrary units, respectively) and carboxyl esterase activities (5055 +/- 1789, 4080 +/- 752, 1713 +/- 522 mU/mg, respectively). IC20 CPT-11 prior to 5-FU was additive to synergistic in SW620, HT-29 and SNU-C4 cells (CIs of 0.7 +/- 0.1). By contrast, pre-treatment with IC20 5-FU antagonised the CPT-11 mediated growth inhibition (CIs of 1.9 +/- 0.4, 1.7 +/- 1.1, 2.5 +/- 0.9, respectively). Simultaneous drug treatment did not produce more cell growth inhibition than either drug alone in the SW620 and the HT-29 cells, but was additive or antagonistic in the SNU-C4 cells (CIs of 1.1 +/- 0.3 and 2.2 +/- 1.4), depending on the ratio of the drugs. Increased DNA damage in the SW620 and HT-29 cells was only seen when IC20 CPT-11 preceded IC50 5-FU, resulting in approximately 40 and 25%, respectively, more lesions than for IC50 5-FU alone. In the SNU-C4 cells, not only such a treatment, but also simultaneous drug treatment produced (30 to 60%) more DNA damage than either drug alone. Our results show clear sequence-dependent antiproliferative effects and DNA damage formation by CPT-11 and 5-FU at combinations of low, fixed doses with higher, varying doses in cultured human colon carcinoma cells, and may be of relevance to the design of improved chemotherapeutic regimens in this disease. PMID- 10674004 TI - Treatment of normal and malignant cells with nucleoside analogues and etoposide enhances deoxycytidine kinase activity. AB - Deoxycytidine kinase (dCK), one of the rate-limiting enzymes in the intracellular metabolism of many antileukaemic drugs, was shown to be stimulated after treatment of human tonsillar lymphocytes by 2-chloro-2'-deoxyadenosine (cladribine, CdA) (Sasvari-Szekely, et al., Biochem Pharmacol 1998, 56, 1175 1179). Here we present a comparative study of different normal and malignant cells in respect to the activation of dCK by CdA. G-phase lymphocytes showed a higher sensitivity for dCK stimulation than S-phase cells. Normal and leukaemic peripheral blood mononuclear cells, as well as the promyelocytic cell line HL60 responded to CdA treatment by a 2-5-fold increase in activity of dCK. However, no significant stimulation was detected either in CCRF-CEM T-lymphoblastoid cells, or in K562 myeloid cells. Thymidine kinase (TK) activity was not stimulated in any cases. Treatment of these cells with several other analogues beside CdA, such as 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA), 2-fluoro-1-beta-D arabinosyladenine (Fludarabine, FaraA) and 1-beta-D-arabinosylcytosine (cytarabine, araC) gave similar results to CdA treatment. Enhancement of dCK activity could also be achieved with the topoisomerase II inhibitor, etoposide. In contrast, 2-chloro-riboadenosine (CrA) had no effect on the dCK at concentrations of 10 microM or less, while dCyd and 5-aza-dCyd caused slight inhibition. These results indicate that treatment of cells with several inhibitors of DNA synthesis potentiates the dCK activity. The drugs widely differ in their stimulatory effect on dCK, and there are also 'responsive' and 'non responsive' cells with respect to dCK activation. Thus, enhancement of the dCK activity by specific drugs in 'responsive' cells might give a rationale for combination chemotherapy. PMID- 10674005 TI - Tamoxifen added to adjuvant chemotherapy in premenopausal women with early breast cancer: is it standard practice or still a subject for study? PMID- 10674006 TI - Selective oestrogen receptor modulation: molecular pharmacology for the millennium. AB - Knowledge of the mechanism of action and pharmacology of tamoxifen and raloxifene, for the prevention of breast cancer and osteoporosis respectively, has opened the door for the discovery of multifunctional medicines. There is now the potential to prevent osteoporosis, coronary heart disease, breast and endometrial cancer in postmenopausal women with elevated risk factors. PMID- 10674007 TI - Should chemotherapy be used as a treatment of advanced colorectal carcinoma (ACC) in patients over 70 years of age? PMID- 10674008 TI - Selection of markers to predict tumour response or survival: description of a novel approach. PMID- 10674009 TI - Alcohol intake and late-stage promotion of breast cancer. AB - Breast cancer risk in women rises with increasing alcohol intake and is widely assumed to be mediated by increased oestrogen concentrations. However, observations that mechanisms and risk are likely to differ between pre- and postmenopausal women suggest that the postmenopausal disease in particular, may involve a promoting role for concomitants of hyperinsulinaemia which is commonly associated with alcoholic cirrhosis of the liver. The MEDLINE database and ongoing studies were examined for clinical, epidemiological and laboratory data on; (a) alcohol-related increase in the incidence of breast cancer in relation to menopausal status, oestrogen concentrations and the oestrogen receptor (ER) status of the tumour; (b) activation of insulin-like growth factor 1 receptor (IGF1R) in mammary tissue by alcohol-related hyperinsulinaemia; (c) interaction between ER and IGF1R in breast cancer cell systems. Epidemiological association between alcohol intake and increased breast cancer risk is more clearly seen in postmenopausal than premenopausal women, and a significant risk is associated with intake of more than two drinks (over 30 g) daily over a period of years. Alcohol-related hyperinsulinaemia is reported to increase with increasing degrees of cirrhosis and damage to liver function. Laboratory evidence suggests that hyperinsulinaemia can stimulate expression of IGF1R in mammary tissue, and this protein is likely to have a crucial role in mitogenesis and transformation to an oestrogen-independent malignant phenotype. It is postulated that in women with a history of long-term intake of moderate quantities of alcohol, the concomitants of hyperinsulinaemia may help to stimulate progression in precancerous breast lesions in the years leading up to the menopause and may increase the risk of breast cancer manifesting after the menopause. PMID- 10674010 TI - Tamoxifen in high-risk premenopausal women with primary breast cancer receiving adjuvant chemotherapy. Report from the Danish Breast Cancer co-operative Group DBCG 82B Trial. AB - Following modified radical mastectomy, pre- and perimenopausal (amenorrhoea for < 5 years) patients with stage II or III breast cancer received CMF (cyclophosphamide 600, methotrexate 40, 5-fluorouracil 600 mg/m2 intravenously (i.v.) every 4 weeks, 9 cycles). The effect on recurrence-free survival (RFS) and overall survival (OS) of the addition of adjuvant tamoxifen (TAM) to adjuvant chemotherapy was examined by randomisation either to no additional treatment (n = 314), or concurrently TAM 30 mg daily for 1 year (n = 320). 40% had positive, 12% negative and 48% unknown receptor status. One year after surgery 21% versus 35% (CMF + TAM versus CMF) were still menstruating (P < 0.01). With a median follow up of 12.2 years there was no difference in RFS (10-year RFS 34% versus 35%, P = 0.81) or OS (45% versus 46%, P = 0.73). In a Cox proportional hazards model, tumour size, number of metastatic lymph nodes, frequency of metastatic nodes in relation to total number of nodes removed, degree of anaplasia, age, and menostasia within the first year after operation were significant independent prognostic factors for RFS, and the same factors except age for OS. No significant interactions with TAM were seen. Thus, in this group of pre- and perimenopausal high-risk early breast cancer patients with heterogeneous receptor status given CMF i.v., concurrent TAM for 1 year did not improve the outcome. These results do not exclude that receptor positive patients may benefit from adjuvant TAM for longer periods given sequentially to chemotherapy. PMID- 10674011 TI - Caring About Women and Cancer (CAWAC): a European survey of the perspectives and experiences of women with female cancers. AB - This paper reports on the findings of the largest ever European survey of female patients' perceptions of their cancer treatment. It has provided clarification of what women consider important in relation to their management and has identified several areas where more research is needed. It has shown that women's knowledge about cancer before diagnosis is poor and the number undergoing regular screening could be improved. Women are not being adequately prepared and educated about what to expect from treatment and steps should be taken as a matter of urgency to redress this shortcoming. It was revealed that whilst families were the primary source of support to female cancer patients, women also derive considerable support from healthcare professionals, particularly senior doctors; more attention should be paid by specialists and nurses to developing psychological skills to cope with this. In this context, further research is needed into how support groups may best meet patient needs. PMID- 10674012 TI - Differential prognosis of replication error phenotype and loss of heterozygosity in sporadic colorectal cancer. AB - Several distinct genetic alterations have been associated with colorectal tumorigenesis. This study investigated the frequency of microsatellite instability, also known as replication error (RER), and loss of heterozygosity (LOH) at six chromosome regions in sporadic colorectal cancer (CRC). Eighty-six tumour and paired normal mucosa samples were included in the study. A polymerase chain reaction (PCR)-based technique was performed to analyse six (CA)n dinucleotide repeats located near or within regions containing important genes implicated in the complex process of colorectal tumorigenesis (chromosomes 2p, 3p, 5q, 11p, 17p and 18q). Overall, LOH frequency was higher in RER-tumours (25/46, 54.3%) compared with RER+ tumours (9/40, 22.5) (P = 0.04). To investigate prognostic implications, survival analysis was performed for 66 patients. Compared with RER- tumours, patients with RER+ tumours at 2p, 3p, 5q, 11p or 18q were found to have an improved prognosis (overall survival, P = 0.02 and disease free survival (DFS) P = 0.005) this variable being an independent prognostic factor by multivariate analysis (P = 0.001). Overall survival of patients whose tumours were LOH+ was significantly shorter compared with those without LOH (overall survival, P = 0.008 and DFS, P = 0.01). Thus, tumours displaying RER+ and LOH+ phenotype, as established by microsatellite analysis, show a differential prognosis. These data indicate that this may be a useful tool for the identification of patients at different risks affected by CRC. PMID- 10674013 TI - TP53 gene mutation status in pretreatment biopsies of oesophageal adenocarcinoma has no prognostic value. AB - Identification of markers which help to predict response to treatment and overall survival at the time of diagnosis would assist in the management of patients with oesophageal adenocarcinoma. In the present study we investigated the prognostic significance of mutations to the TP53 tumour suppressor gene in a large, consecutive series of oesophageal adenocarcinomas. The incidence of TP53 mutation determined by molecular analysis of endoscopic biopsy specimens was 36% (49/135). No statistically significant difference was observed in patient survival according to the TP53 status of the tumour biopsy. The median survival time for patients with mutation was 12 +/- 1 months compared with 14 +/- 2 months for patients with TP53. These results demonstrate that mutation of the TP53 gene is not a useful predictive marker for patient survival in oesophageal adenocarcinoma. PMID- 10674014 TI - Decreased toxicity and increased efficacy of cancer chemotherapy using the pineal hormone melatonin in metastatic solid tumour patients with poor clinical status. AB - Melatonin (MLT) has been proven to counteract chemotherapy toxicity, by acting as an anti-oxidant agent, and to promote apoptosis of cancer cells, so enhancing chemotherapy cytotoxicity. The aim of this study was to evaluate the effects of concomitant MLT administration on toxicity and efficacy of several chemotherapeutic combinations in advanced cancer patients with poor clinical status. The study included 250 metastatic solid tumour patients (lung cancer, 104; breast cancer, 77; gastrointestinal tract neoplasms, 42; head and neck cancers, 27), who were randomized to receive MLT (20 mg/day orally every day) plus chemotherapy, or chemotherapy alone. Chemotherapy consisted of cisplatin (CDDP) plus etoposide or gemcitabine alone for lung cancer, doxorubicin alone, mitoxantrone alone or paclitaxel alone for breast cancer, 5-FU plus folinic acid for gastro-intestinal tumours and 5-FU plus CDDP for head and neck cancers. The 1 year survival rate and the objective tumour regression rate were significantly higher in patients concomitantly treated with MLT than in those who received chemotherapy (CT) alone (tumour response rate: 42/124 CT + MLT versus 19/126 CT only, P < 0.001; 1-year survival: 63/124 CT + MLT versus 29/126 CT only, P < 0.001). Moreover, the concomitant administration of MLT significantly reduced the frequency of thrombocytopenia, neurotoxicity, cardiotoxicity, stomatitis and asthenia. This study indicates that the pineal hormone MLT may enhance the efficacy of chemotherapy and reduce its toxicity, at least in advanced cancer patients of poor clinical status. PMID- 10674015 TI - Bone mineral density in children with acute lymphoblastic leukaemia. AB - Bone mineral density (BMD) may be negatively affected by the disease or its treatment in patients with acute lymphoblastic leukaemia (ALL). Therefore, we evaluated lumbar spine and total body BMD and bone metabolism in children with ALL at diagnosis, during treatment with chemotherapy and 1 year after completion of treatment. 32 children (21 boys and 11 girls) participated in the study. 14 children started the study at diagnosis and 18 during or after the treatment period. Lumbar spine and total body BMD were measured with dual energy X-ray absorptiometry, and expressed as standard deviation scores (SDS). Blood samples were obtained to assess bone metabolism. 3 of 14 children had low lumbar spine BMD (< -2 S.D.) at diagnosis. All children had normal total body BMD. Markers of bone turnover were depressed. Total body BMD SDS decreased significantly during the first year of treatment (P < 0.001). Lumbar spine BMD SDS did not change significantly. Parameters of bone turnover increased to normal during the treatment period. Parathyroid hormone had increased significantly after 1 year (P < 0.05). Mineral homeostasis was disturbed in some patients during treatment. 4 of 9 patients had low total body BMD and 1 patient low lumbar spine BMD one year after completion of treatment. All patients had normal biochemical results at that time. In conclusion, lumbar spine BMD and bone turnover were decreased in some patients at diagnosis. Total body BMD decreased significantly during treatment and was low in 4 of the 9 patients 1 year after completion of the treatment. PMID- 10674016 TI - Local therapy and other factors influencing site of relapse in patients with localised Ewing's sarcoma. United Kingdom Children's Cancer Study Group (UKCCSG). AB - Relapse patterns have been documented in 191 children with localised Ewing's sarcoma treated with the United Kingdom Children's Cancer Group (UKCCSG) Ewing's Tumour regimen ET2. All received chemotherapy comprising ifosfamide, vincristine, doxorubicin and actinomycin D. Local treatment modality was excision and or radiotherapy depending on tumour site and response to primary chemotherapy. Although not strictly comparable, due to the clinical indications used for each modality, local relapse rates were very low and were similar, irrespective of the type of local treatment modality: radiotherapy (3/56), surgery (7/114) or a combination (0/20). Combined relapse (local + distant) rates were similarly low irrespective of the type of local therapy: radiotherapy (4/56), surgery (4/114) or a combination (0/20). Overall survival was lower in females (P = < 0.04), older children (P = < 0.002) and those with primaries at sites other than long bones (P = < 0.02). It is concluded that with effective intensive chemotherapy combined with either radiotherapy or surgery, local control in this study was excellent at sites other than the pelvis. Preventing distant relapse, predominantly to lung and bone, remains the major challenge. PMID- 10674017 TI - Epidemiological aspects of soft tissue sarcomas (STS)--consequences for the design of clinical STS trials. AB - The purpose of the study was to gain insight into epidemiological aspects of soft tissue sarcomas (STS), based on the population-based cancer registry of the Comprehensive Cancer Center North-Netherlands (CCCN), and to provide data for the development of future STS clinical trials. 456 primary STS (Kaposi, urogenital and gastro-intestinal STS excluded), registered from 1989 to 1995 by the cancer registration of the Comprehensive Cancer Center North-Netherlands (CCCN), were analysed. The annual, age-adjusted, STS incidence was 3.6 per 100,000. Incidence increased with age. Half of the patients were over the age of 65 years. Malignant fibrous histiocytomas and liposarcomas were most frequently encountered. At presentation, nodal involvement was rare (3-8%). Distant metastases were more frequently encountered (9-14%), and appeared to be related to tumour size and site. Above 70 years of age, 16% of patients received no treatment at all, especially for metastatic disease. PMID- 10674018 TI - Descriptive epidemiology of soft tissue sarcomas in Vaud, Switzerland. AB - Trends in incidence and survival from soft tissue sarcomas (STS) were analysed for the period 1974-1994 using data from the Cancer Registry of the Swiss canton of Vaud. A total of 645 cases were registered. The most common histotypes were fibrosarcoma (0.82/100,000 males, 0.86/100,000 females, world standard, 1990 1994), leiomyosarcoma (0.90/100,000 males, 1.28/100,000 females, 1990-1994), and Kaposi's sarcoma (3.10/100,000 males in 1990-1994). Overall incidence rates for STS increased from 2.68/100,000 males in 1974-1979 to 6.86 in 1990-1994, and from 3.61 to 4.27 in females. However, after excluding Kaposi's sarcoma, no consistent trend over time was observed, peak rates (approximately 4.40/100,000) being registered in the late 1980s for both sexes, with some levelling off thereafter. Five-year relative survival was 17% for Kaposi's sarcomas, and 51% for other STS (all STSs, 45%). These data indicate that there has been no major new risk factor for STS of such a relevance to modify appreciably the overall rates on a population level, except from the impact of the AIDS epidemic for Kaposi's sarcoma. PMID- 10674019 TI - Vitamin D analogues suppress IGF-I signalling and promote apoptosis in breast cancer cells. AB - Survival factors are known to promote cell viability, and factor deprivation can be a potent apoptotic signal. Insulin-like growth factors are potent mitogens and inhibitors of apoptosis for many normal and neoplastic cells with insulin-like growth factor-I (IGF-I) being the most effective in many breast cancer cell lines. 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and its analogues inhibit IGF-I stimulated growth of MCF-7 human breast cancer cells. The aim of this study was to determine the relationship between inhibition of IGF-I responsiveness and induction of apoptosis by vitamin D analogues in breast cancer cells. Vitamin D analogues EB1089 and CB1093 inhibited autonomous and IGF-I-stimulated growth of MCF-7 and T47D cells and autonomous growth of IGF-I-insensitive Hs578T cells. In MCF-7 cells, IGF-I alone (4 nM) protected against apoptosis mediated by serum deprivation. Co-treatment with vitamin D analogues prevented the anti-apoptotic effects of IGF-I. In T47D cells, IGF-I treatment provided only partial protection against apoptosis induced by serum deprivation and co-incubation of serum deprived cells with 100 nM CB1093 and IGF-I abrogated this partial protection. In Hs578T cells, addition of IGF-I did not prevent apoptosis induced by serum deprivation. However, treatment with CB1093 attenuated the protective effect of the serum in these cells. Our findings suggest that vitamin D analogues inhibit IGF-I signalling pathways to promote apoptosis in breast cancer cells. PMID- 10674020 TI - Mutations in hMSH6 alone are not sufficient to cause the microsatellite instability in colorectal cancer cell lines. AB - Microsatellite instability (MSI) at simple repeated sequences characterises a distinct mechanism of carcinogenesis in hereditary nonpolyposis colorectal cancer (HNPCC), as well as sporadic colorectal cancers displaying MSI. Such MSI is associated with mutations of hMSH2 and hMLH1, and somatic frameshift mutations in TGF-beta RII, IGFIIR, BAX, hMSH3 and hMSH6 at simple repeated sequences in coding regions. The aim of this study was to look for a correlation between mutations in mismatch repair genes and frameshift mutations in colorectal cancer cell lines with MSI. Using 22 colorectal cancer cell lines, we examined the MSI status at mononucleotide repeat microsatellite markers and mutations in hMSH2 and hMLH1, TGF-beta RII, IGFIIR, BAX, hMSH3 and hMSH6. Thirteen of 22 lines (59%) displayed MSI. In these 13 lines showing MSI, 10 lines (77%) had mutations in TGF-beta RII, nine lines (69%) in BAX, seven lines (54%) in hMSH6, and six lines (46%) in hMSH3, while mutations in the IGFIIR gene were identified in only two lines (15%). Of the 13 lines with MSI, six lines (46%) harboured mutations/deletions in hMSH2 (two nonsense mutations, three deletions and one no expression of transcripts) and three of these cell lines (50%) had mutations both in the hMSH2 and hMSH3 genes. Two cell lines (15%) had mutations/deletions in hMLH1 (one missense mutation and one deletion) and these two cell lines also had mutations in hMSH3. One line had a mutation in hMSH3 only, although this line showed MSI and had mutations in TGF-beta RII, IGFIIR and BAX. All lines with mutations in hMLH1, hMSH2, TGF-beta RII, IGFIIR, BAX and hMSH3 genes showed MSI. However, of the nine lines without MSI, two (22%) had homozygous mutations in hMSH6. In these two cell lines, no mutations were identified in hMLH1, hMSH2, TGF-beta RII, IGFIIR, BAX and hMSH3. Our results indicate that mutations in hMLH1, hMSH2 and hMSH3 are associated with MSI, but mutations in hMSH6 are not. We conclude that mutations in hMSH6 alone are not sufficient to cause MSI, although protein functional effects of these mutations should still be examined. PMID- 10674021 TI - Loss of heterozygosity at chromosome 13q in hepatocellular carcinoma: identification of three independent regions. AB - Loss of heterozygosity (LOH) on chromosome 13q is one of the most common genetic alterations in hepatocellular carcinoma (HCC) and might be involved in liver cancer development through inactivation of tumour suppressor genes. In order to narrow down the region of 13q loss, we examined the pattern of loss of heterozygosity (LOH) in tumours from 88 HCC patients, using 18 microsatellite markers on 13q. Thirty-eight of the 88 tumours (43%) showed LOH for at least one marker. Of these, two tumours (5%) showed 13q whole arm allelic loss, while the remaining 36 tumours (95%) had partial allelic loss. The LOH pattern defined by the 36 tumours suggested the existence of at least three different smallest common deleted regions which might be involved in the carcinogenesis of HCC. The first, the most centromeric in the 13q12.3 is, close to the BRCA2 gene, defined by D13S171; the second, the most telomeric region in the 13q31-32 band, is defined by D13S154 and D13S157; the third, the intermediate region at 13q14.3, which is near the RB gene, is defined by loci D13S268. The rate of LOH at 13q31 32 was significantly higher in Hepatitis B-surface antigen (HBsAg)-positive patients than HBsAg-negative HCC patients, pointing to a candidate gene related to the development of HBsAg-positive HCCs. PMID- 10674022 TI - Suppressive subtractive hybridisation reveals differential expression of serglycin, sorcin, bone marrow proteoglycan and prostate-tumour-inducing gene I (PTI-1) in drug-resistant and sensitive tumour cell lines of haematopoetic origin. AB - The development of therapy-induced drug resistance is still one of the most important therapeutic limitations. Nevertheless, an integrating view of the molecular mechanisms underlying resistance development in general is missing. In order to shed some light on the network of this resistance development, we established drug-resistant (doxorubicin (DX), methotrexate (MTX), cisplatin (cisPt), vincristine (Vin)) derivatives of six tumour cell lines (Jurkat, U937, HL60, DoHH-2, K562 and ARH77) of haematopoetic origin. Differential gene expression of drug-sensitive parental cell lines and the drug-resistant derivatives thereof was analysed by suppressive subtractive hybridisation. After dot blot screening for differential expression and sequencing of the cloned PCR fragments, differential expression was confirmed by Northern blot analysis. In an attempt to discriminate for differentially expressed genes only related to one or the other of the investigated drugs, the cDNAs of various resistant sublines (doxorubicin-, methotrexate-, cisplatin-resistant Jurkat cells) were pooled and compared with the sensitive parental cell line. In addition, cDNAs of the resistant derivatives of the different haematopoetic tumour cell lines were pooled and compared with the pooled cDNAs of the corresponding sensitive haematopoetic cell lines to eliminate cell line to cell line variations that were not related to drug resistance. As a result of this screening, the following genes showed a higher (at least 2-fold) or exclusive expression in the drug resistant variants: serglycin, sorcin, BMPG (bone marrow proteoglycan gene) and PTI-1 (prostate-tumour-inducing gene 1). In addition, elevated expression of hsp90, previously found by our group to be upregulated in the drug-resistant colon carcinoma cell line LoVo H67P was found to be overexpressed in drug resistant HL60 cells. PMID- 10674023 TI - Comments on: Tagging sentinel lymph nodes: a study of 100 patients with breast cancer. Bobin, et al., Eur J Cancer 1999, 35, 569-573. PMID- 10674024 TI - Detection of drug misuse--an addictive challenge. AB - It is now accepted that drug misuse is a large and growing problem in the United Kingdom, some estimates of the number of regular illicit drug users being as high as three million. The aim of this paper is to provide insight into the methods used to detect drug misuse. The strategy adopted by one laboratory is described and methods of screening for, and identification of, a wide range of compounds are provided. No claim is made that this is the best approach or that the list of drugs detected is comprehensive; the range of drugs encountered is always increasing and the lists are constantly updated. It is hoped that users of toxicology laboratory services will gain an appreciation of the capabilities and limitations of the techniques available; and that those who may wish to provide such a service will find the necessary information in a readily accessible format. PMID- 10674025 TI - Biological consequences of the BCR/ABL fusion gene in humans and mice. PMID- 10674026 TI - Next of kin clinics: a new role for the pathologist. PMID- 10674027 TI - Detection of telomerase in hepatocellular carcinomas using a PCR ELISA assay: comparison with hTR expression. AB - BACKGROUND: While telomerase is undetectable in most normal somatic tissues, telomerase activation has been detected in many immortal cell lines and various cancers. AIM: To investigate telomerase expression in hepatocellular carcinoma, and to assess the expression of the RNA component of telomerase, hTR. METHODS: 39 hepatocellular carcinomas were studied using a telomerase polymerase chain reaction (PCR) enzyme linked immunosorbent assay, which does not require radioactive PCR amplification and yields a semiquantitative measurement. Expression of hTR was also assessed by a non-radioactive in situ hybridisation procedure. The correlations between these two markers and the clinicopathological data were analysed. RESULTS: Telomerase activity was detected in 23 of 39 hepatocellular carcinoma specimens (59%). Comparison of hepatocellular carcinoma with and without telomerase expression, or with high and low telomerase (10 cases v 13 cases), showed no differences in the principal clinicopathological data. Although median survival was lower in the group with detectable telomerase activity than in that with undetectable activity (510 v 720 days) the difference was not significant (log-rank test, p = 0.08). hTR expression was detected in 11 of 14 cases of hepatocellular carcinoma tested (78%) and in four of 12 samples of adjacent non-cancerous tissue (33%). Five tumours and four non-cancerous tissues were positive for hTR, whereas no telomerase activity was detected in these. CONCLUSIONS: The presence of telomerase activity in hepatocellular carcinomas is confirmed. No correlation was observed between clinicopathological data and telomerase expression in hepatocellular carcinoma, but survival seemed better in the absence of telomerase expression. hTR seems to be more widely expressed than telomerase. PMID- 10674028 TI - Expression of CD44 on bile ducts in primary sclerosing cholangitis and primary biliary cirrhosis. AB - AIM: To examine expression of CD44, a transmembrane glycoprotein involved in lymphocyte homing and activation, in inflammatory liver diseases. METHODS: Formalin fixed, paraffin embedded tissues were obtained from normal, uninvolved liver from patients undergoing partial hepatectomy for metastatic carcinoma (9) and transplant hepatectomy specimens from patients with primary biliary cirrhosis (12), primary sclerosing cholangitis (8), autoimmune hepatitis (3), hepatitis C (3), and secondary sclerosing cholangitis (1). Expression of CD44 (using antibodies to three core epitopes), HLA-DR, and lymphocyte phenotypic markers was studied by immunohistochemistry. RESULTS: CD44 expression was not detected in either hepatocytes or biliary epithelial cells in normal livers. In sections from all 27 transplant hepatectomy specimens, CD44 was positive in bile duct epithelial cells but not in hepatocytes. The proportion of CD44+ ducts was much higher in biliary disease than in chronic hepatitis. By contrast, expression of HLA-DR was detected in a relatively small percentage of bile ducts. Activated, memory phenotype CD4+ T lymphocytes were increased in the parenchyma of all diseased livers and an infiltrate of activated CD8+ cells within the biliary epithelium was evident in inflammatory biliary disease. CONCLUSIONS: CD44 appears to play an important role in the development of autoimmune biliary disease by promoting lymphoepithelial interactions, whereas HLA-DR may be involved in the subsequent progression of these conditions. PMID- 10674029 TI - Vascular endothelial growth factor mRNA expression in minimal change, membranous, and diabetic nephropathy demonstrated by non-isotopic in situ hybridisation. AB - AIM: To investigate vascular endothelial growth factor (VEGF) mRNA expression in glomerular disease in the context of heavy proteinuria. METHODS: Non radioisotopic in situ hybridisation was performed using a cocktail of 12 deoxyoligonucleotides complementary to VEGF mRNA labelled during solid phase synthesis with 2,4-dinitrophenyl. Archival renal biopsies were studied from cases of minimal change nephropathy, membranous nephropathy, diabetic nephropathy, and controls, matched for age, sex, race, and storage time. Hybrid detection used NBT/BCIP colorimetric development. RESULTS: More VEGF mRNA positive glomerular cells per unit cross sectional diameter were seen in minimal change nephropathy (mean (SEM), 19.35 (1.5)) compared with controls (12.6 (1.73)), p < 0.01. In contrast, fewer were seen in diabetic nephropathy (5.93 (0.97)) compared with controls (9.97 (1.25)), p < 0.03. Analysis of membranous nephropathy (10 (1.62)) showed no difference from controls (10.98 (1.51)), NS. In addition, in minimal change nephropathy there was a significant correlation between 24 hour protein excretion at the time of biopsy and the number of VEGF mRNA cells per glomerulus (r = 0.08, p = 0.01). CONCLUSIONS: Using non-radioisotopic in situ hybridisation, VEGF mRNA is almost exclusively expressed by visceral glomerular epithelial cells. Abnormal numbers of cells are seen in both minimal change and diabetic nephropathy. As VEGF exists in a number of functionally distinct isoforms, further study of qualitative VEGF isoform expression in diagnostic groups is indicated. PMID- 10674030 TI - Autoimmune markers are undetectable in end stage idiopathic dilated cardiomyopathy. AB - BACKGROUND: Autoreactive humoral and cellular immune responses may be involved in the pathogenesis of idiopathic dilated cardiomyopathy (IDC). Certain human leucocyte antigens (HLA) could also be linked to the development of IDC. AIM: To determine whether various markers of autoimmunity are present in the final phase of the disease, to substantiate the role of an autoimmune process in IDC. METHODS: 37 patients with end stage IDC were studied, together with 39 patients with end stage heart disease of known aetiology who were included for comparison. Multiple myocardial tissue samples from the explanted heart of each patient were evaluated (immuno)histologically. An indirect immunofluorescence assay was used to screen patient serum samples for the presence of heart specific autoantibodies. HLA class I and II frequencies were determined in each group and compared with HLA frequencies from healthy blood donors. RESULTS: Only scanty small mononuclear cell infiltrates were present in myocardial tissue of seven patients with IDC and of 11 patients with heart disease of known cause. The majority of these inflammatory cells were negative for T cell markers. All blood specimens were negative for heart specific autoantibodies and there was no apparent association of IDC with particular HLA phenotypes. CONCLUSIONS: These findings suggest that an active autoimmune process is not involved in the end stage of IDC. PMID- 10674031 TI - The effects of freeze drying and freeze drying additives on the prothrombin time and the international sensitivity index. AB - AIM: To determine whether freezing, freeze drying protective additives, or freeze drying of plasma samples from patients on coumarin treatment and from normal individuals affects prothrombin times or the international sensitivity index (ISI) calibration. METHODS: The effect of the addition of the protective additives singly and combined on the prothrombin time of coumarin samples and normal samples before and after freeze drying was observed using high and low ISI reference thromboplastins. ISI values were also determined. RESULTS: Freezing caused a prolongation of prothrombin time in the normal plasma samples with both reagents, which was significant with the low ISI human. Prolongation (non significant) of the prothrombin time in coumarin plasma samples occurred with the human reagent only. Significant prolongation of normal prothrombin time by some of the protective additives before and after freeze drying was observed with both thromboplastins but to a greater extent with the human. Significant prolongation of prothrombin time in coumarin plasma samples was observed, but again was more marked with human thromboplastin. An approximate ISI was determined on the 20 coumarin samples. The only marked ISI change was with the WHO human thromboplastin after freeze drying of plasma, where a decrease from 0.95 to 0.90 was observed, corresponding to a marked prothrombin ratio increase. CONCLUSIONS: Freeze drying additives and the freeze drying procedure prolong normal and coumarin prothrombin times, with low ISI thromboplastin. Less marked prolongations occurred with a high ISI rabbit reagent, coumarin samples showing more significant prolongations. Marked ISI change in freeze dried plasma was only recorded with the low ISI ECAA human reagent. Frozen normal plasma samples cannot be used with confidence for ISI calibrations. PMID- 10674032 TI - Postmortem prolactin as a marker of antemortem stress. AB - AIM: To determine whether raised prolactin concentrations could be identified using postmortem venous blood and whether the level of prolactin correlated with antemortem stress. METHODS: Blood was obtained from the right femoral vein of 100 random adult necropsy cases, all of whom had been dead less than four days. Prolactin was determined in the samples by microparticle immunoassay. The levels of prolactin obtained were then analysed in relation to sex and cause of death, with particular emphasis on a history of antemortem stress and drug use. RESULTS: Prolactin in all cases of trauma was in the normal range (up to 500 mU/l). In cases of sudden unexpected deaths the mean concentration was 533 mU/l (95% confidence interval (CI), 372 to 694 mU/l). Postoperative deaths or cases with chronic disease had a mean value of 1027 mU/l (95% CI, 735 to 1319 mU/l). Cases of suicide had a mean value of 1398 mU/l. Analysis of the suicides by sex showed a significant difference, the mean in female cases being 2072 mU/l compared with 692 mU/l in male cases. In three of the four female suicides with the highest prolactin, the hyperprolactinaemia might have been attributable to a drug effect, but one case still had unexplained hyperprolactinaemia. CONCLUSIONS: It is possible to detect prolactin reliably in postmortem venous blood samples. Prolactin values at necropsy differ according to the cause of death, with markedly higher values in postoperative deaths and in the chronically ill. Hyperprolactinaemia in cases of suicide is likely to result from the effects of the drugs used, but the levels were higher than previously reported. PMID- 10674033 TI - Age related prevalence of hepatitis G virus in South Africans. AB - AIM: To investigate the age related prevalence of hepatitis G virus (HGV) infection and its mode of transmission in relation to hepatitis B (HBV) and C (HCV) co-infection in South African blacks. METHODS: Reverse transcriptase polymerase chain reaction was performed to detect active infection, using primers for the 5'-NCR, NS5a, and NS3 regions. Antibodies to HGV envelope-2 protein (anti E2), which measures past infection, were also sought. RESULTS: The overall prevalence of active infection was 116/580 (20%). A higher prevalence was noted in HBsAg carriers (28/106; 26.4%) and HCV positive subjects (25/82; 30.5%). In contrast to developed countries, active and past infection was seen in 12.9% and 12.1% of the general population, respectively (subjects negative for HBsAg and anti-HCV markers and with normal alanine aminotransferase values), with a total prevalence of 21.1% (52/248). Viraemia and anti-E2 were almost mutually exclusive. The distribution of viraemia by age was: < or = 15 years, 20/223 (9.0%); 16-35 years, 42/147 (28.6%); > or = 36 years, 37/151 (24.5%), with a significant difference (p = 0.001) in age related prevalence. A similar trend was observed for the prevalence of past infection in the general population. CONCLUSIONS: HGV infection begins in childhood and increases with age in South Africa, but transmission is largely independent of HBV and HCV. No association was found between HGV viraemia and hepatitis, or with co-infection with either HBV or HCV. PMID- 10674035 TI - Image sampling in static telepathology for frozen section diagnosis. AB - BACKGROUND: A frozen section diagnostic service is often not directly available in small rural or mountain hospitals. In these cases, it could be possible to provide frozen section diagnosis through telepathology systems. Telepathology is based on two main methods: static and dynamic. The former is less expensive, but involves the crucial problem of image sampling. AIMS: To characterise the differences in image sampling for static telepathology when undertaken by pathologists with different experience. METHODS: As a test field, a previously studied telepathology method based on multimedia email was adopted. Using this method, three pathologists with different levels of experience sampled images from 155 routine frozen sections and sent them to a distant pathology institute, where diagnoses were made on digital images. After the telepathology diagnoses, the glass slides of both the frozen sections and the definitive sections were sent to the remote pathologists for review. RESULTS: Four of 155 transmissions were considered inadequate by the remote pathologist. In the remaining 151 cases, the telepathology diagnosis agreed with the gold standard in 146 (96.7%). There was no significant divergence between the three pathologists in their sampling of the images. Each case comprised five images on average, acquired in four minutes. The overall time for transmission was about 19 minutes. CONCLUSIONS: The results suggest that in routine frozen section diagnosis an inexperienced pathologist can sample images sufficiently well to permit remote diagnosis. However, as expected, the internet is too unreliable for such a time dependent task. An improvement in the system would involve integrated real time features, so that there could be interaction between the two pathologists. PMID- 10674034 TI - Oesophageal mesenchymal tumours: clinicopathological features and absence of Epstein-Barr virus. AB - BACKGROUND: Recent studies have suggested that the Epstein-Barr virus (EBV) is associated with smooth muscle tumours (leiomyoma and leiomyosarcoma) in patients with human immunodeficiency virus and in organ transplant recipients. Leiomyoma is the most common mensenchymal tumour found in the oesophagus. AIM: To report a single institution experience on oesophageal mesenchymal tumours and to determine whether EBV is associated with these tumours. METHODS: 40 sporadic oesophageal mesenchymal tumours were studied and their diagnosis confirmed on pathological review and immunohistochemical studies. Formalin fixed, paraffin was embedded tissues from these tumours were analysed for EBV using in situ hybridisation for two messenger RNA (mRNA) probes, EBER and BamH1 W. RESULTS: The oesophageal mesenchymal tumours comprised 36 leiomyomas, two undifferentiated stromal tumours, and two gastrointestinal autonomic nerve tumours (GANTs). Median age of the patients with leiomyoma (26 men, 10 women) was 62 years (range 30 to 85) and 81% of them had an asymptomatic lesion. The median longitudinal size was 1.2 cm. Multiple leiomyomas were seen in 11% of the patients and calcification was noted in one tumour. Coexisting squamous cell carcinoma was found in one third of cases. The stromal tumours were small, asymptomatic, and located in the lower third of the oesophagus, while the GANTs were large, symptomatic, and found in the upper third of the oesophagus. EBV mRNAs were not detected in all these tumours. CONCLUSIONS: The clinicopathological features of oesophageal leiomyoma, undifferentiated stromal tumour, and GANT were different. Some oesophageal leiomyomas were associated with oesophageal squamous cell carcinomas. EBV is not associated with sporadic oesophageal mesenchymal tumours. PMID- 10674036 TI - Rapid detection of the factor V Leiden (1691 G > A) and haemochromatosis (845 G > A) mutation by fluorescence resonance energy transfer (FRET) and real time PCR. AB - A rapid method based on fluorescence resonance energy transfer (FRET) and real time polymerase chain reaction (PCR) was used to identify the haemochromatosis genotype in 112 individuals and the factor V genotype in 134 individuals. The results were compared with conventional methods based on restriction enzyme digestion of PCR products. The two methods agreed in 244 of the 246 individuals; for the other two individuals, sequencing showed that they had been incorrectly genotyped by the standard method but correctly genotyped by FRET. The simplicity, speed, and accuracy of real time PCR analysis using FRET probes make it the method of choice in the clinical laboratory for genotyping the haemochromatosis and factor V genes. PMID- 10674037 TI - A latex slide agglutination test for rapid detection of antimyeloperoxidase antibody. AB - AIM: To develop and test a new latex slide agglutination test (MPO-LSAT) to detect antimyeloperoxidase (anti-MPO) antibody in serum. METHODS: Latex bead coating was adjusted to give maximum sensitivity by attending to latex size, MPO to latex ratio for coupling, ratio of diluted serum to MPO-latex, reaction time and temperature for coupling, and reaction time for agglutination. Inhibition studies were performed using MPO, proteinase 3, bactericidal/permeability increasing protein, and lactoferrin. RESULTS: There was very good correlation between this test and the conventional anti-MPO enzyme linked immunosorbent assay (ELISA): 81% of sera positive in the ELISA were positive by MPO-LSAT. MPO-LSAT results correlated better with IgM anti-MPO than with IgG anti-MPO. CONCLUSIONS: MPO-LSAT is a simple diagnostic test that is potentially useful in the clinical laboratory as a rapid screening tool for vasculitic diseases. PMID- 10674038 TI - Review of the clinical activity of medical microbiologists in a teaching hospital. AB - BACKGROUND: The clinical interactive role of medical microbiologists has been underestimated and the discipline is perceived as being confined to the laboratory. Previous studies have shown that most microbiology interaction takes place over the telephone. AIM: To determine the proportion of clinical ward based and laboratory based telephone interactions and specialties using a microbiology service. METHODS: Clinical microbiology activity that took place during November 1996 was prospectively analysed to determine the distribution of interactions and specialties using the service. RESULTS: In all, 1177 interactions were recorded, of which nearly one third (29%) took place at the bedside and 23% took place on call. Interactions involving the intensive treatment unit, general ward visits, and communication of positive blood cultures and antibiotic assays were the main areas of activity identified. There were 147 visits to 86 patients on the general wards during the study, with the number of visits to each individual varying from one to eight. The need for repeated visits reflected the severity of the underlying condition of the patients. Ward visits were regarded as essential to obtain missing clinical information, to assess response to treatment, and to make an appropriate entry in a patient's notes. CONCLUSIONS: Ward visits comprise a significant proportion of clinical microbiology interactions and have potential benefits for patient management, service utilisation, and education. PMID- 10674039 TI - The use of histological techniques for the demonstration of ion exchange resins. AB - AIM: To establish the staining characteristics of certain ion exchange resins in histological material, with a view to enabling confident differential identification. METHODS: Various histological staining procedures were applied to selected pathological material and prepared agar blocks containing the cation exchange resin calcium polystyrene sulphonate and the anion exchange resin cholestyramine. RESULTS: Calcium polystyrene sulphonate uniquely stained strongly by a direct Schiff's reagent procedure without any preoxidation and by the Ziehl Neelsen method. Cholestyramine was negative by the former method but stained strongly with a standard Congo red technique. CONCLUSIONS: These staining results are consistent with the known structure and properties of polystyrene sulphonate and cholestyramine resins. Polystyrene sulphonate resins have the virtually pathognomonic feature of direct Schiff positivity, while morphology, location, and strong non-birefringent Congo red positivity facilitate the identification of cholestyramine. It is possible that the intrinsic staining characteristics of cholestyramine may be lost once it has bound to its target. PMID- 10674040 TI - Severe hypoglycaemia caused by raised insulin-like growth factor II in disseminated breast cancer. AB - A 60 year old woman with disseminated ductal carcinoma of the breast developed non-islet cell tumour induced hypoglycaemia (NICTH). The concentrations of the two insulin-like growth factors, IGF-I and IGF-II, were < 2 nmol/l and 94.1 nmol/l, respectively. The IGF-II to IGF-I ratio was > 47 (normal < 10). Insulin (< 3 mu/l) and C peptide (< 100 pmol/l) were both undetectable. PMID- 10674041 TI - Association of HLA-DRB1 alleles with giant cell tumour of bone. AB - AIM: To examine the possible influence of the MHC class II antigens alleles in the formation of the multinucleate aggressive giant cell tumour of bone. METHODS: HLA class II antigen alleles were investigated in eight white patients from north east England with confirmed diagnosis of giant cell tumour of bone. All had locally aggressive, immunophenotypically HLA-DR negative giant cell tumours. RESULTS: Five of the eight patients were found to be positive for HLA DRB1*0801/3, the distribution of this allele in healthy white controls being quite low (2%). All but one of the patients possessing DRB1*080 also expressed DRB1*070. CONCLUSIONS: HLA-DRB1*080 is pre-dominant in patients with immunophenotypic HLA-DR negative giant cell tumour of bone; individuals with the genotype 070/080 are at particularly high risk of developing giant cell tumour of bone. PMID- 10674042 TI - Differential expression of high molecular weight caldesmon in colorectal pericryptal fibroblasts and tumour stroma. AB - AIM: To investigate an extent of smooth muscle differentiation of pericryptal fibroblasts. METHODS: Expression of high molecular weight caldesmon (h-CD) and alpha smooth muscle actin was investigated in 123 invasive colorectal adenocarcinomas and their surrounding non-neoplastic tissues. RESULTS: The monoclonal antibody to h-CD, which showed predominantly positive immunostaining in well differentiated smooth muscle cells, recognised pericryptal fibroblasts, smooth muscle cells, and pericytes, but was not reactive to myofibroblasts. Antibody to alpha smooth muscle actin recognised not only pericryptal fibroblasts, smooth muscle cells, and pericytes but also myofibroblasts. CONCLUSIONS: Pericryptal fibroblasts show greater smooth muscle differentiation than myofibroblasts and there is a possibility that they are well differentiated smooth muscle cells; h-CD is an excellent marker to discriminate pericryptal fibroblasts from myofibroblasts. PMID- 10674043 TI - Follow up of women with borderline cervical smears as defined by national guidelines. AB - AIM: To determine the proportion of women with abnormalities in cervical smears corresponding to borderline nuclear change, as defined by national guidelines, which return to normal or persist as cytological or histological abnormalities. METHODS: 313 women with borderline nuclear change diagnosed by a single pathologist using the national criteria were followed up for up to two years. RESULTS: On initial follow up, 45% of women had a negative smear or biopsy, 46.5% had a low grade cytological or histological abnormality, and 8.5% had a high grade abnormality. Of 81 patients in whom a second follow up smear or biopsy was available, 47% had no detectable abnormality, 38.5% had low grade lesion, and 14.5% had a high grade lesion. In total, 32 patients (10.2%) had a high grade lesion (defined as moderate or severe dyskaryosis on smear or CINII or CINIII on biopsy) on at least one follow up sample. CONCLUSIONS: The results support the use of the national criteria defining borderline nuclear change in identifying women at increased risk of developing a high grade cervical intraepithelial neoplasia, as identified histologically or cytologically, and highlight the importance of follow up in these patients. PMID- 10674044 TI - Cytokeratin 10/13, 14, 7, 8, and 18 in invasive squamous cell carcinoma and adenocarcinoma of the uterine cervix. PMID- 10674045 TI - No pain, no gain? PMID- 10674046 TI - Support of clinical trials. PMID- 10674047 TI - If the WCB can do it, why not others? PMID- 10674048 TI - Real-world effectiveness of antihypertensive drugs. PMID- 10674049 TI - Why aren't we falling for anticoagulant therapy? PMID- 10674050 TI - Non-heart-beating organ donation. PMID- 10674051 TI - To pay or not to pay? A decision and cost-utility analysis of angiotensin converting-enzyme inhibitor therapy for diabetic nephropathy. AB - BACKGROUND: Angiotensin-converting-enzyme (ACE) inhibitor therapy can significantly delay the progression of diabetic nephropathy to end-stage renal failure (ESRF). The main obstacle to successful compliance with this therapy is the cost to the patients. The authors performed a cost-utility analysis from the government's perspective to see whether the province or territory should pay for ACE inhibitors for type I diabetic nephropathy on the assumption that cost is a major barrier to compliance with this important therapy. METHODS: A decision analysis tree was created to demonstrate the progression of type I diabetes with macroproteinuria from the point of prescription of ACE inhibitor therapy through to ESRF management, with a 21-year follow-up. Drug compliance, cost of ESRF treatment, utilities and survival data were taken from Canadian sources and used in the cost-utility analysis. One-way and two-way sensitivity analyses were performed to test the robustness of the findings. RESULTS: Compared with a no payment strategy, provincial payment of ACE inhibitor therapy was found to be highly cost-effective: it resulted in an increase of 0.147 in the number of quality-adjusted life-years (QALYs) and an annual cost savings of $849 per patient. The sensitivity analyses indicated that the cost-effectiveness depends on compliance, effect of benefit and the cost of drug therapy. Changes in the compliance rate from 67% to 51% could result in a swing in cost-effectiveness from a savings of $899 to an expenditure of more than $1 million per additional QALY. A 50% reduction in the cost of ACE inhibitors would result in a cost savings of $299 per additional QALY with compliance rates as low as 58% in the provincial payment strategy. INTERPRETATION: Provincial coverage of ACE inhibitor therapy for type I diabetes with macroproteinuria improves patient outcomes, with a decrease in cost for ESRF services. PMID- 10674053 TI - Telemedicine and fetal ultrasonography in a remote Newfoundland community. PMID- 10674052 TI - Blood recipient notification for hepatitis C in Prince Edward Island. AB - BACKGROUND: Two of the major risk factors for hepatitis C are injection drug use and receipt of blood or blood products. Many patients are unaware that they have received transfusions. In 1998 Prince Edward Island conducted a province-wide look-back notification program to notify patients who had received transfusions in PEI between Jan. 1, 1984, and June 1, 1990. The authors present the results of the notification program. METHODS: A registry for recipients of blood and blood products was created from the province's Red Cross blood bank records. The registry data were linked with Vital Statistics data to determine death status and with Health Registration data to determine residence status of recipients (in PEI or moved out of province). All identified recipients with a current PEI mailing address were sent a letter recommending hepatitis C virus (HCV) testing. Laboratory records were checked to determine HCV test results. RESULTS: The registry contained data for 6086 recipients of blood or blood products during the look-back period; 51.1% (3109/6086) had died by the time of notification. Of the remainder, 18.4% (549/2977) were not directly notified because they had moved out of province, had refused delivery of the notification letter or had died recently, or because identifying information was missing from the blood bank records. Of the recipients who were notified 80.4% (1953/2428) underwent testing, and 2.2% (43/1953) were found to be HCV positive. Most of these (58.1% [25/43]) had undergone testing before notification. The HCV positivity rate differed significantly between recipients tested before notification and those tested after notification (9.9% v. 1.1%, p < 0.001). HCV-positive recipients were more likely than other notified recipients to have had multiple transfusions (39.5% v. 9.5%, p < 0.001). INTERPRETATION: Before notification 4.1% of PEI recipients had undergone HCV testing. After notification 91.2% of PEI recipients were identified as tested, dead or moved out of province. The notification program resulted in the identification of the majority of PEI's transfusion-related cases of hepatitis C. PMID- 10674054 TI - The seroprevalence of hepatitis A and B in people testing positive for hepatitis C. PMID- 10674055 TI - The need for greater involvement of regulatory agencies in assessing adverse drug reactions. PMID- 10674056 TI - HIV testing of patients: let's waive the waiver. PMID- 10674057 TI - Scientific harassment by pharmaceutical companies: time to stop. PMID- 10674058 TI - The diagnosis and management of insomnia in clinical practice: a practical evidence-based approach. AB - Insomnia, or the dissatisfaction with the quantity, quality or timing of sleep, is a common complaint. Because the definition of "normal" sleep is not well established, the estimates of the prevalence and severity of insomnia vary widely. Insomnia is often secondary to underlying psychiatric and medical conditions, and these should be evaluated and treated as a first measure. Nonpharmacological interventions for insomnia including sleep hygiene manoeuvres and exercise are recommended, although the success of these interventions has not been well documented. Benzodiazepines have been the pharmacologic agents of choice for the treatment of insomnia, but there is reason to exercise caution with their use; their overall benefit compared with placebo appears to be minor, and they are often associated with adverse cognitive effects. Unfortunately, no other class of drugs has proven to be superior to the benzodiazepines in terms of benefit:risk ratio. Given the importance of sleep for health and normal daily functioning the diagnosis, prognosis and treatment of insomnia should be a research priority. PMID- 10674060 TI - Chasing the dragon--neurological toxicity associated with inhalation of heroin vapour: case report. PMID- 10674061 TI - A view inside the womb. PMID- 10674059 TI - Meta-analysis of benzodiazepine use in the treatment of insomnia. AB - OBJECTIVE: To systematically review the benefits and risks associated with the use of benzodiazepines to treat insomnia in adults. DATA SOURCES: MEDLINE and the Cochrane Controlled Trials Registry were searched for English-language articles published from 1966 to December 1998 that described randomized controlled trials of benzodiazepines for the treatment of insomnia. Key words included "benzodiazepines" (exploded), "randomized controlled trial" and "insomnia." Bibliographies of relevant articles were reviewed for additional studies and manufacturers of benzodiazepines were asked to submit additional randomized controlled trial reports not in the literature. STUDY SELECTION: Articles were considered for the meta-analysis if they were randomized controlled trials involving patients with insomnia and compared a benzodiazepine with placebo or another active agent. Of the 89 trials originally identified, 45 met our criteria, representing a total of 2672 patients. DATA EXTRACTION: Data were extracted regarding the participants, the setting, details of the intervention, the outcomes (including adverse effects) and the methodologic quality of the studies. DATA SYNTHESIS: The meta-analyses of sleep records indicated that, when compared with placebo, benzodiazepines decreased sleep latency by 4.2 minutes (non-significant; 95% confidence interval (CI -0.7 to 9.2) and significantly increased total sleep duration by 61.8 minutes (95% CI 37.4 to 86.2). Patient reported outcomes were more optimistic for sleep latency; those randomized to benzodiazepine treatment estimated a sleep latency decrease of 14.3 minutes (95% CI 10.6 to 18.0). Although more patients receiving benzodiazepine treatment reported adverse effects, especially daytime drowsiness and dizziness or light headedness (common odds ratio 1.8, 95% CI 1.4 to 2.4), dropout rates for the benzodiazepine and placebo groups were similar. Cognitive function decline including memory impairment was reported in several of the studies. Zopiclone was not found to be superior to benzodiazepines on any of the outcome measures examined. INTERPRETATION: The use of benzodiazepines in the treatment of insomnia is associated with an increase in sleep duration, but this is countered by a number of adverse effects. Additional studies evaluating the efficacy of nonpharmacological interventions would be valuable. PMID- 10674062 TI - Mechanical ventilation: what's new when your patient is blue? PMID- 10674063 TI - Can Quebec afford dialysis for every 80-year-old patient? PMID- 10674064 TI - London MDs provide window to surgery's future. PMID- 10674065 TI - Reform party rejects physician's reforms. PMID- 10674066 TI - Are Canadians aware of the risks of smoking? PMID- 10674067 TI - Population rises dramatically but number of med-school applicants remains stagnant. PMID- 10674068 TI - Mycobacterium ulcerans disease; Buruli ulcer. PMID- 10674069 TI - Manson Lecture. Meningococcal meningitis in Africa. AB - This review covers the history of meningococcal meningitis in Africa since epidemics of the infection were first described around 100 years ago. It is possible that an epidemic strain of the meningococcus was introduced into West Africa from the Sudan by pilgrims returning from the Haj around the turn of the century. Since 1905 major epidemics of meningococcal meningitis have occurred in countries of the Sahel and sub-Sahel every few years, culminating in a massive epidemic in which nearly 200,000 cases were reported in 1996. Attempts to control epidemic meningococcal meningitis in Africa by vaccination with meningococcal polysaccharide vaccines have met with only modest success because epidemics can progress with great rapidity and vaccination is often started too late. This situation should be improved as a result of a recent initiative, the International Coordinating Group (ICG), which is contributing to better surveillance in countries at risk and ensuring that vaccine is available when needed. However, in the medium term, the best prospect for the control of meningococcal meningitis in Africa lies in the recent development of polysaccharide-protein conjugate vaccines which, unlike polysaccharide vaccines, are immunogenic in the very young, induce immunological memory and are likely to give long-lasting protection. PMID- 10674070 TI - The work of the Tropical Health and Education Trust (THET) in training for healthcare. Introduction. PMID- 10674071 TI - Delivering care for diabetes in Ethiopia. PMID- 10674072 TI - The Nottingham City Hospital-Jimma Institute of Health Sciences link. PMID- 10674073 TI - Training trainers to improve laboratory services. PMID- 10674074 TI - The development of professional and postgraduate education through collaborative links with Kumasi, Ghana. PMID- 10674075 TI - Emergency life-saving surgical courses in Africa. PMID- 10674076 TI - Seasonal and nocturnal domiciliary human landing/biting behaviour of Lutzomyia (Lutzomyia) evansi and Lutzomyia (Psychodopygus) panamensis (Diptera; Psychodidae) in a periurban area of a city on the Caribbean coast of eastern Venezuela (Barcelona; Anzoategui State). AB - In recent years, in addition to American cutaneous leishmaniasis (ACL), a significant number of cases of American visceral leishmaniasis (AVL) have been reported in periurban areas of Barcelona city (Anzoategui State, Venezuela). We studied the bionomics of Lutzomyia (Lutzomyia) evansi and Lutzomyia (Psychodopygus) panamensis, possible vectors of AVL and ACL, respectively, in El Rincon, a periurban village of that city. To evaluate the seasonal domiciliary landing/biting activity of sandflies on human bait, a house was chosen in El Rincon. Landing catches were carried out between 18:00 and 06:00, once a month for a year. The results show the presence of 2 species, Lu. (Lu.) evansi (89.9%) and Lu. (Psy.) panamensis (10.1%). Lu. evansi was most abundant in the months of October and July, associated with the bimodal cycle of annual rainfall in the area. Maximum landing/biting activity of Lu. evansi was observed at 24:00 and 03:00. These findings suggest that at this time of the year and at these hours there is heightened risk of the transmission of AVL. Lu. panamensis monthly abundance also shows a direct association with rainfall and maximum landing/biting activity was observed between 02:00 and 03:00. The lower domiciliary abundance of Lu. panamensis suggests its greater importance in the extradomiciliary transmission of ACL. PMID- 10674077 TI - Similar blackfly attraction by onchocerciasis patients and individuals putatively immune to Onchocerca volvulus. AB - In areas endemic for onchocerciasis, a small number of individuals show no detectable infection with Onchocerca volvulus despite an apparently similar exposure to the transmitting blackflies. Such individuals have been termed putatively immune. Since several studies have indicated marked host differences in attractiveness for blood-seeking insects, putative immunity in O. volvulus infection may result, at least in part, from low vector attractiveness of the respective individuals. In an area hyperendemic for onchocerciasis (Guinea), where Simulium yahense is the predominant vector, we organized fly catches by putatively immune individuals and individuals with moderate-to-high worm counts. No differences were found between the 2 groups with respect to (i) the attraction of blackflies, (ii) the attraction of blackflies infected with O. volvulus, or (iii) the numbers of O. volvulus larvae carried by the attracted blackflies. PMID- 10674078 TI - Cryptococcal meningitis in AIDS patients: an emerging opportunistic infection in Senegal. PMID- 10674079 TI - Biased distribution of msp1 and msp2 allelic variants in Plasmodium falciparum populations in Thailand. AB - Plasmodium falciparum isolates were obtained from Thai patients attending a malaria clinic on the Thai-Kampuchean border over 4 cross-sectional surveys carried out at 3-monthly intervals. The genetic structure of the parasite populations was determined by nested polymerase chain reaction (PCR) amplification of polymorphic regions of 3 P. falciparum antigen genes: msp1, msp2 and glurp. Although a high degree of diversity characterized these isolates, the overall population structure of the parasites associated with patent malaria infections was observed to remain relatively stable over time. The highest degree of polymorphism was observed with msp2, and the mean number of lines per infection (multiplicity of infection) calculated with this marker was higher than that obtained using msp1 or glurp alone, or combined. Infections with > or = 2 parasite lines were seen in 76% of the samples, and were proportionally more numerous at the start and end of the rainy season. Two interesting exceptions to the random distribution were observed and involved 2 allelic variants which in one case were found dissociated (msp1 MAD20-family) and in the other were associated (msp2 FC27-family). The epidemiological significance of these types of data is discussed. PMID- 10674081 TI - Comparative susceptibility of two members of the Anopheles oswaldoi complex, An. oswaldoi and An. konderi, to infection by Plasmodium vivax. AB - We compared the susceptibility of Anopheles oswaldoi and An. konderi to infection by Plasmodium vivax based on the proportion of mosquitoes presenting oocysts and sporozoites. Anophelines were captured in the State of Acre and Rondonia, Brazilian Amazon, and used to obtain F1 progenies. After emergence of adults, male genitalia of mosquitoes of each family were dissected in order to identify them as either An. oswaldoi or An. konderi. F1 progenies of field-captured An. oswaldoi, An. konderi and An. darlingi (used as control) were fed simultaneously on P. vivax-infected blood. Mosquitoes were dissected on day 10-12 after feeding and examined for the presence of oocysts and sporozoites. Both An. oswaldoi and An. konderi developed oocysts in the midguts, however, the percentage of oocyst positive mosquitoes for An. oswaldoi (13.8%) was higher than for An. konderi (3.3%), and only An. oswaldoi developed salivary infection with sporozoites (6.9% of positivity). Infection rates in An. darlingi ranged from 22.5% to 30.0% for both oocysts and sporozoites. These results indicate that An. oswaldoi can transmit P. vivax and suggest that it is more susceptible than An. konderi. Although An. oswaldoi is an exophilic and zoophilic species, it may be involved in malaria transmission as possibly occurred in the State of Acre. PMID- 10674080 TI - A cohort study of Plasmodium falciparum diversity during the dry season in Ndiop, a Senegalese village with seasonal, mesoendemic malaria. AB - Prolonged carriage of Plasmodium falciparum in humans during the dry season is critical for parasite survival, as the infected subjects constitute a major reservoir in the absence of transmission. Yet, very little is known about the host/parasite interactions contributing to parasite persistence. In order to study the characteristics of P. falciparum infections during the dry season, we have genotyped parasites collected from untreated, asymptomatic individuals during 3 cross-sectional surveys conducted during the dry season in Ndiop, a Senegalese village with seasonal, mesoendemic malaria. Monthly entomological surveillance did not detect any transmission during that period. Parasite prevalence decreased markedly in the children aged < 7 years after 7 months of undetected transmission, but was stable in older children and adults throughout the dry season. In all chronically infected individuals, infection complexity remained stable, but there were substantial fluctuations of individual genotype(s), reflecting complex dynamics of multiple-clone infections during chronic asymptomatic parasite carriage. This fluctuation resulted in changes in the msp1 and msp2 allelic distribution within the cohort after 7 months of undetected transmission, contrasting with the stability observed during the preceding rainy season in that village. PMID- 10674082 TI - Is Leishmania infantum zymodeme MON-253 involved in an outbreak among intravenous drug users? PMID- 10674083 TI - School-based schistosomiasis control programmes: a comparative study on the prevalence and intensity of urinary schistosomiasis among Nigerian school-age children in and out of school. AB - A cross-sectional study was conducted in February 1998 on the prevalence and intensity of urinary schistosomiasis among school-age children in and out of school at Adim village in Nigeria to test the objective of delivering a control programme through the school system. School enrollment figures and non-attendance rate were collated from questionnaires that were self-administered by heads of families. Prevalence and intensity of infection were determined following filtration of urine and counting of carbol fuchsin-stained eggs of Schistosoma haematobium. The rates of regular school attendance, irregular attendance and non attendance were 69.1%, 5.1%, and 25.8%, respectively. These indices were not significantly associated with the age of the schoolchildren (P > 0.05). Boys (76.6%) were more associated with regular attendance than girls (61.4%) (P < 0.0001) while girls had a higher rate of non-attendance (32.7%) than males (19.1%) (P < 0.0001). Although more out-of-school children were infected (90.7%) than those in school (86.8%), the difference was not statistically significant (P > 0.05). The same association was established in the variation of mean egg count between the 2 study populations though intensity was higher among out-of-school children. The principal reasons proffered for the high rate of non-attendance listed in their order of importance were: economic, sickness, poor performance, refusal, farming and fishing. A dual method of control that would in incorporate the integration of recognized local authorities is suggested in areas with moderate school attendance rate like Adim, as lack of treatment of infected out of-school children ensures continuous contamination and re-infection. PMID- 10674084 TI - Differences in tuberculosis incidence rates in township and in rural populations in Ntcheu District, Malawi. AB - There has been a large upsurge of tuberculosis (TB) in many countries in sub Saharan Africa, mainly as a result of the co-existing human immunodeficiency virus (HIV) epidemic. Malawi has had a well-run National TB Control Programme (NTP) with good registration and recording of cases. For some years the NTP has had the impression that TB in the country is concentrated around townships and is less prevalent in the rural areas. This impression was investigated in a rural district (Ntcheu District) in Malawi. Data on new TB cases were collected from the district TB register for the years 1992-96 and average annual TB incidence rates per 100,000 for semi-urban and rural populations were calculated for this period. There was a significantly higher incidence of TB, particularly amongst cases with smear-negative pulmonary TB and extrapulmonary TB, in the semi-urban population compared with the rural population. Possible explanations could be higher HIV seroprevalence rates in semi-urban areas compared with rural areas, under-diagnosis at health centres or poor access to medical facilities for rural people. PMID- 10674085 TI - Evaluation of a standardized leishmanin skin test in Guatemala. AB - Before recommending the skin-test use at national level in Guatemala of an antigen prepared from Leishmania major (a Leishmania species not found in the New World), we conducted a study in 100 Guatemalans to determine its sensitivity and specificity. The antigen consisted of 0.1 mL of a solution that contained 5 x 10(6) promastigotes of L. major (MRHO/IR/75/VAX). Positive leishmanin skin test (LST) reactions at 48 h were observed in 16 (80%) of 20 patients with proven active cutaneous leishmaniasis (CL), 18 (90%) of 20 with previously treated proven CL, and in 18 (90%) of 20 with a history and compatible scan of previously suspected but unconfirmed CL. None of 20 healthy controls or 20 patients with skin lesions due to causes other than CL had positive reactions to the LST, giving a sensitivity of 85% and specificity of 100%. There were no statistically significant differences in ethnic group, age, duration of the lesion, lesion size or Leishmania species between the 34 persons with true positive reactions. Even though it will be necessary to test this antigen on a larger number of patients, these preliminary results show that this antigen is specific and reasonably sensitive in identifying current or past CL and that it is a reasonable choice for epidemiological studies on CL in Guatemala. PMID- 10674086 TI - Use of molecular tools for the diagnosis and typing of a Leishmania major strain isolated from an HIV-infected patient in Burkina Faso. PMID- 10674087 TI - Increased sensitivity of routine laboratory detection of Strongyloides stercoralis and hookworm by agar-plate culture. AB - The efficacy of agar-plate culture has been evaluated for the detection of Strongyloides stercoralis and hookworm, compared with direct smear, the formalin ether sedimentation technique and the filter-paper method. Of 1085 stool samples from the routine laboratory service at King Chulalongkorn Memorial Hospital in Bangkok, 241 samples harboured S. stercoralis, 153 hookworm and 2 Rhabditis hominis. The recovery rate of S. stercoralis by agar-plate culture is significantly superior to the other methods (P < 0.005). The ratios of positive results from the methods used to the total number of S. stercoralis-positive cases were as follows: 1:1.03 by agar-plate culture, 1:1.85 by the filter-paper method, 1:1.98 by the sedimentation technique and 1:10.48 by direct stool smear. A similar trend of the efficacy ratio of each method was obtained for hookworm detection. The characteristic furrows left by hookworm larvae, and larvae and adults of S. stercoralis could be used for preliminary species identification. Daily search for furrows on agar plates for up to 6 consecutive days resulted in an increased sensitivity for diagnosis of both S. stercoralis and hookworm infections. PMID- 10674088 TI - Detection of antibodies against hepatitis A virus in eluates of blood spotted on filter-paper: a pilot study in Rio de Janeiro, Brazil. AB - The validity of blood spotted on to filter-paper (BSOFP) eluates for the detection of antibodies against hepatitis A virus (HAV) was investigated in 718 individuals (children and adults) during a field study in a small area in Rio de Janeiro State, Brazil. Serum samples were considered the 'gold standard'. BSOFP eluates were analyzed by 2 different techniques: microplate competitive enzyme linked immunosorbent assay (ELISA) of the whole study group and microparticle enzyme immune assay (MEIA) of a subsample of 59 individuals. For BSOFP eluates by ELISA, sensitivity and specificity were 89.6% (95% CI: 84.7-93.1) and 97.5% (95% CI: 95.6-98.7), respectively. For a seroprevalence of anti-HAV antibodies of 32%, the positive predictive value was 94.5% (95% CI: 90.3-97.0) and the negative predictive value was 95.2% (95% CI: 92.8-96.8). The test efficiency was 95.0% (95% CI: 93.1-96.4). Similar results were found for BSOFP eluates by MEIA. Agreement between the 2 techniques used for BSOFP (ELISA and MEIA) was also high (kappa = 0.93). These results encourage the more widespread application of BSOFP as a means of surveillance for large-scale epidemiological studies for hepatitis A. PMID- 10674089 TI - The optimum relative centrifugal force and centrifugation time for improved sensitivity of smear and culture for detection of Mycobacterium tuberculosis from sputum. AB - Direct microscopy is the only available method for diagnosis of tuberculosis in most centres in developing countries. Methods to improve the sensitivity of direct smear are an urgent requirement. Sputum specimens artificially seeded with known concentrations of Mycobacterium tuberculosis were liquefied and decontaminated with sodium hydroxide-sodium citrate-N-actyl-L-cysteine solutions. They were subjected to different centrifugation forces and centrifugation times after which the centrifuged deposits were examined by smear and culture. Statistical analysis of results was carried out using EpiInfo version 6.0. The optimum relative centrifugal force (RCF) and centrifugation time combination was 4000 g for 15 min. The sensitivity of detection at an RCF of 4000 g for 15 min was 5000 organisms/mL and 500 organisms/mL for smear and culture, respectively. When results of 163 clinical samples were analyzed after centrifugation at 4000 g for 15 min sensitivity of the direct smear improved from 63% to 92% (P < 0.05) and negative predictive value from 30.5% to 45% (P < 0.05) when culture was considered the 'gold standard'. With the concentrated smear there was a reduction in specificity from 82% to 60% (P > 0.05). As most laboratories are equipped with a simple centrifuge, smear sensitivity can be improved with this simple modification. The other advantage is that the same centrifuged deposit can be cultured, in contrast to when sodium hypochlorite is used for liquefaction. PMID- 10674090 TI - Does the use of urinary reagent strip tests improve the bedside diagnosis of meningitis? PMID- 10674091 TI - Blood in the urine of adolescent girls in an area of Ghana with a low prevalence of infection with Schistosoma haematobium. PMID- 10674092 TI - Acute attacks in the extremities of persons living in an area endemic for bancroftian filariasis: differentiation of two syndromes. AB - The natural history of lymphatic disease in human filariasis remains unclear, but recurrent episodes of acute lymphangitis are believed to constitute a major risk factor for the development of chronic lymphoedema and elephantiasis. Prospective analysis of 600 patients referred to the filariasis clinic of the Centro de Pesquisas Aggeu Magalhaes/FIOCRUZ in Recife, Brazil, indicated that 2 distinct acute syndromes accompanied by lymphangitis occur in residents of this filariasis endemic area. One syndrome, which we call acute filarial lymphangitis (AFL), is caused by the death of adult worms. It is relatively uncommon in untreated persons, usually is asymptomatic or has a mild clinical course, and rarely causes residual lymphoedema. The second syndrome, of acute dermatolymphangioadenitis (ADLA), is not caused by filarial worms per se, but probably results from secondary bacterial infections. ADLA is a common cause of chronic lymphoedema and elephantiasis in Recife as well as in other areas of Brazil where lymphatic filariasis is not present. The syndromes of AFL and ADLA can be readily distinguished from each other by simple clinical criteria. PMID- 10674093 TI - Fatal primary dengue infections in Brazil. PMID- 10674094 TI - Haematological characteristics and HIV status of pregnant women in Abidjan, Cote d'Ivoire, 1995-96. AB - To describe the haematological profile of pregnant women and to compare these characteristics according to HIV serostatus in Abidjan, Cote d'Ivoire, a cross sectional study was made in the context of a research intervention programme to reduce mother-to-child transmission (MTCT) of HIV (ANRS 049 trial). HIV testing was systematically proposed to pregnant women attending the mother and child health clinic of a community health centre. Blood samples were tested for HIV antibodies using Genelavia and Peptilav. The haematological parameters were measured with a Coulter counter. From May 1995 to March 1996, 1646 pregnant women accepted HIV testing and had a full blood count available. The prevalence of HIV infection was 12.0% (n = 197). The prevalence of anaemia (haemoglobin [Hb] < 11 g/dL) was 70.1%, n = 1155 (95% confidence interval 68-72%) and significantly higher in HIV+ (81.7%, n = 161) than in HIV- women (68.9%, n = 994) (P < 0.001). Severe anaemia (Hb < 7 g/dL) was present in 1.9% of the women (n = 31), 4.6% (n = 9) in HIV+ and 1.5% (n = 22) in HIV- women (P < 0.001). HIV infection, primigravidae and secundigravidae were factors independently associated with anaemia. Anaemia was highly prevalent in this population while severe anaemia was rare. HIV infection was a contributor to anaemia in pregnancy. As zidovudine, with its known haematological toxicity, has recently been introduced to prevent MTCT of HIV in developing countries, screening HIV+ women for severe anaemia is necessary. PMID- 10674095 TI - Maternal iron status and intrauterine growth retardation. AB - The objective of this study was to compare the iron status of 356 mother-baby pairs who had intrauterine growth retardation (IUGR) with 356 mother-baby pairs who had appropriate weight for gestational age (AGA). Mothers were selected in 1991/92 from 4 hospitals in Campinas city, Brazil, where 95% of deliveries take place. Gestational age of the newborns was determined by the Capurro method. Newborns were classified as having IUGR according to the Lubchenco birthweight for gestational age standard. Haemoglobin (Hb), haematocrit (Ht) and ferritin were determined, respectively, by the cyanmethaemoglobin method, an haematocrit centrifuge, and an immunoenzymetric assay. Mean levels of Hb and Ht were higher in IUGR (16.4 g/dL; 51.7%) than in AGA babies (15.7 g/dL; 49.7%) (P < 0.001), as a consequence of intrauterine hypoxia. Higher maternal levels of ferritin (> 50 micrograms/L) were more common in IUGR than in AGA mothers (P < 0.001). Forty seven percent (335/712) of the IUGR and AGA mothers were anaemic (Hb < or = 11.0 g/dL), but only 4.4% (31/356) of them had low levels of ferritin (< or = 10 micrograms/L). We advise further large epidemiological studies involving IUGR and AGA mother-baby pairs, elucidating the mechanisms underlying the plasma-volume changes in pregnancy, and the prevalence of iron-deficiency anaemia assessed by different indicators, in view of the fact that ferritin can be affected by inflammation and infection, important risk factors for IUGR in developing countries. PMID- 10674096 TI - Alopecia induced by ants. PMID- 10674097 TI - The effect of artesunate combined with standard antimalarials against chloroquine sensitive and chloroquine-resistant strains of Plasmodium falciparum in vitro. AB - The interactions of artesunate with chloroquine, mefloquine, quinine, doxycycline and pyrimethamine were tested in vitro against chloroquine-sensitive (D10) and chloroquine-resistant (RSA11) strains of Plasmodium falciparum. Mefloquine and quinine both showed synergism of artesunate activity against each of the strains, whilst doxycycline showed an additive interaction. Pyrimethamine combinations were antagonistic, and the combination of artesunate with chloroquine was antagonistic against RSA11, and additive against D10. Although weak antagonism in vitro might not indicate any clinical significance, synergism with artesunate may increase the clinical usefulness of either drug, and could potentially be of value in delaying the emergence of resistance. PMID- 10674098 TI - Treatment of vivax malaria on the western border of Thailand. AB - The efficacy of chloroquine (25 mg base/kg over 3 days) in Plasmodium vivax malaria was evaluated in 1995/96 in 342 patients living in an endemic area on the western border of Thailand. Clearance of fever and parasites was obtained within 2 days in > 95% of the patients, and all were aparasitaemic by day 4. Reappearance of P. vivax occurred in 1 patient on day 21 and in 8 by day 28, giving a 28-day cure rate of 97% [95% confidence interval (CI) 95-99%]. By day 63, the relapse/re-infection rate was 63% (95% CI 57-69%). Most reappearances of parasitaemia (85%; 121/143) were symptomatic. These patients were retreated either with chloroquine alone (n = 70) or with chloroquine and primaquine (0.25 mg/kg daily for 14 days) (n = 43). Only 1 patient (in the chloroquine-only group) had prolonged parasite clearance (D8) and he developed recurrent P. vivax by day 21 suggesting possible recrudescence. The addition of primaquine to chloroquine reduced the risk of having a third vivax episode within 2 months by 96% (95% CI 83-99%). This resulted in a significantly higher haematocrit at day 42 despite a greater decrease in haematocrit during the first week of treatment with chloroquine-primaquine (P = 0.04). Chloroquine remains highly effective on the western border of Thailand and the use of strictly supervised primaquine effectively prevents relapse. The introduction of primaquine on a large scale in an endemic area still requires a long-term risk-benefit assessment which must take into account potential toxicity, low compliance and reductions in the incidence and severity of P. falciparum infections by co-existent P. vivax. PMID- 10674099 TI - Risk factors for treatment failure after melarsoprol for Trypanosoma brucei gambiense trypanosomiasis in Uganda. AB - We evaluated the treatment failure rate among late-stage human African trypanosomiasis (HAT) patients treated with melarsoprol in Arua, northern Uganda, between September 1995 and August 1996, and identified the risk factors for treatment failure. We conducted a retrospective cohort study in October 1998, and performed a survival analysis. A treatment failure was defined as a late-stage HAT patient fully treated with melarsoprol and classified as an HAT case at any follow-up visit within 2 years after treatment. Among 428 patients treated in the study period, 130 (30.4%) were identified as treatment failure within 2 years after discharge. The multivariate analysis showed that patients who experienced treatment failure after melarsoprol were more likely to have been admitted as a relapsing case (relative hazard, RH = 11.15 [6.34-19.61]), and to have been diagnosed with trypanosomes in the lymph nodes (RH = 3.19 [2.10-4.83]) or in the cerebrospinal fluid (CSF) (RH = 1.66 [1.09-2.53]). The risk of treatment failure also increased with the number of cells in the CSF. The treatment failure rate after melarsoprol observed in Arua is greatly above the expected figures of 3-9%. More research is needed to confirm whether it is related to the variation of melarsoprol pharmacokinetics between individuals, or if it is associated with a reduced susceptibility of the trypanosomes to melarsoprol. The study emphasizes the need for second-line drugs to treat patients that have already received one or several full course(s) of melarsoprol. PMID- 10674100 TI - Treatment outcome of patients with smear-negative and smear-positive pulmonary tuberculosis in the National Tuberculosis Control Programme, Malawi. AB - National tuberculosis control programmes (NTPs) in sub-Saharan Africa do not routinely record or report treatment outcome data on smear-negative pulmonary tuberculosis (PTB) patients. Twelve-month treatment outcome on patients with smear-negative PTB registered in all district and mission hospitals in Malawi during the year 1995 was collected, and was compared with 8-month treatment outcome in smear-positive PTB patients registered during the same period. Of 4240 patients with smear-negative PTB, 35% completed treatment, 25% died, 9% defaulted and 7% were transferred to another district with no treatment outcome results available. In 24% of patients treatment cards were lost and treatment outcome was unknown. These results were significantly inferior to those obtained in 4003 patients with smear-positive PTB in whom 72% completed treatment, 20% died, 4% defaulted, 2% were transferred and 1% had positive smears at the end of treatment. These differences between patients with smear-negative and smear positive PTB were similar when analysed by sex and by most age-groups. Higher mortality rates in patients with smear-negative PTB are probably attributable to advanced HIV-related immunosuppression, and higher default and treatment unknown rates probably reflect the lack of attention paid by TB programme staff to this group of patients. As a result of this country-wide study the Malawi NTP has started to record routinely the treatment outcomes of smear-negative TB patients and has set treatment completion targets of 50% or higher for this group of patients. PMID- 10674101 TI - A preliminary study of neopterin as a potential marker for severe dengue virus infection. PMID- 10674102 TI - [Periodically leaking capillaries]. AB - The systemic capillary leak syndrome (Clarkson's syndrome) is a rare idiopathic disorder, characterized by recurrent episodes of hypovolaemic shock, haemoconcentration and hypoalbuminaemia due to a sudden shift of fluid and macromolecules from the intravascular to the interstitial space. A young man is presented in whom recurrent attacks of hypotension and diffuse swelling were initially attributed to staphylococcal toxic shock syndrome. With the additional finding of a monoclonal gammopathy, the diagnosis of systemic capillary leak syndrome was made. Recognition of this syndrome is important, as prophylactic treatment with terbutaline and theophylline may be beneficial in this life threatening syndrome. PMID- 10674103 TI - [Virtual colonoscopy]. AB - Virtual colonoscopy is a fundamentally new imaging technique, in which volumetric CT or MRI data are processed to virtual endoscopic images of the colon. The main application is early detection of colorectal cancer. To date, no data of studies on screening for colorectal cancer are available. The results of clinical studies show comparable results for endoscopy and virtual colonoscopy for the detection of clinically relevant polyps (> or = 1 cm) and better results of virtual colonoscopy than of conventional barium studies. Whether virtual colonoscopy as a minimally invasive technique will gain a place in screening for colorectal cancer depends on several issues such as costs, effectiveness and acceptance by patients. PMID- 10674104 TI - [Immunology in medical practice. XXVII. Vaccines against malaria: new perspectives]. AB - Malaria belongs to the five most important infections in the world and is responsible for 2-3 million deaths each year. A universally active vaccine against Plasmodium falciparum is not available to this day. However, the feasibility of a vaccine is underlined by the results of experimental studies in animal models as well as by the fact that immunity is acquired in humans after a number of infections. Vaccine development is hampered by the complex life cycle of the parasite as well as by the lack of knowledge about the key molecules involved in the cycle. Moreover the parasite shows an extraordinary capacity to evade the immune response. Over the last decade understanding of the immune response has improved; moreover, a number of Plasmodium derived proteins are in the process or on their way to clinical testing as a vaccine. The most recent activities focus on the delineation of the P. falciparum genome as well as on different strategies for vaccination. PMID- 10674105 TI - [Diagnosis and treatment of cholelithiasis]. AB - Most patients with gallbladder stones are asymptomatic and do not require treatment. Biliary colics are the main indication for laparoscopic cholecystectomy: the treatment of choice for gallbladder stones. Dyspepsia is not an indication for treatment of gallstones. The indications for bile salt therapy and extracorporeal lithotripsy are limited. Acute cholecystitis should be treated with cholecystectomy a chaud whereas longstanding cholecystitis is preferably treated with cholecystectomy a froid. Bile duct stones are mainly treated endoscopically during endoscopic retrograde cholangiopancreatography (ERCP): pancreatitis, bleeding and perforation are the main complications. Prior to cholecystectomy, an ERCP is indicated in case of cholangitis, severe pancreatitis, persisting jaundice, bile duct stones on ultrasonography, or the combination of dilated ducts and abnormal liver function tests. After endoscopic stone removal, a cholecystectomy is indicated for patients < 50 years but a 'wait and see' policy is justified in elderly patients. PMID- 10674106 TI - [Cervical smears taken by physicians' assistants are of lesser quality than smears taken by family physicians, but almost as good as the national average]. AB - OBJECTIVE: To compare the quality of cervical smears taken by practice assistants with the quality of smears taken by general practitioners. DESIGN: Retrospective. METHODS: Data were collected on the 1672 smears taken as part of the population screening for cervical cancer and recorded 13 general practices in Limburg in the year 1997. The results of smears taken by practice assistants were compared with those made by general practitioners with regard to the quality indicators 'presence of endocervical cells' and 'absence of Pap-o'. RESULTS: In 12.3% of the 481 cervical smears taken by the 17 practice assistants endocervical cells were absent. This is statistically significantly more than to the results (7.2%) in 1191 smears taken by the 22 general practitioners (relative risk: 1.70: 95% confidence interval: 1.23-2.70). Similar differences, but not statistically significant, were found with regard to Pap-o results and pathological findings. Most of the practice assistants had 1-2 years' experience. The proportion of smears made by practice assistants in which endocervical cells were lacking was reasonable compared with the average of 10% for the Netherlands as a whole. CONCLUSION: The cervical smears taken by practice assistants were of lower quality than those taken by the general practitioners but almost as good as the nationwide mean. PMID- 10674107 TI - [Regional differences in prehospital time delay for patients with acute myocardial infarction; Rotterdam and Groningen, 1990-1995]. AB - OBJECTIVE: To describe pre-hospital delay times in patients with acute myocardial infarction (AMI) in two regions in the Netherlands: Groningen (a region with high mortality for coronary heart disease (CHD)) and Rotterdam (a region with low CHD mortality). DESIGN: Descriptive. METHOD: The pre-hospital treatment delay of AMI patients in Rotterdam in 1990-1991 versus 1993-1995 was compared and also compared between Groningen en Rotterdam (1993-1995). In each region 3 hospitals participated (1 academic, 2 regional). The data were collected with a structured interview within 7 days after onset of symptoms in hospitalized AMI patients (n = 924) or by interviewing relatives of deceased patients (n = 40). The median patient, general practitioner (GP) and ambulance delays were calculated. RESULTS: Total median pre-hospital delay was 2.5 hours (5-95-percentile: 50 min-36 hours). Median patient delay time was shorter in Groningen than in Rotterdam (respectively 30 and 45 min) and the same applied to doctor delay times (respectively 38 and 72 min). In Rotterdam doctor delay time decreased by 23 min between 1990-1991 and 1993-1995. Median ambulance delay was 30 min in Rotterdam and 35 min in Groningen. Total pre-hospital delay times of self referred patients were 32-78 min shorter than those of patients who consulted a GP before admission. CONCLUSION: Reduction of pre-hospital delay in Rotterdam between 1990 1991 and 1993-1995 was due to faster decision time by the GP. The short pre hospital treatment delay in Groningen in hospitalized patients suggests that relatively more AMI patients die outside hospital which may contribute to the high CHD mortality in this region. PMID- 10674108 TI - [Increase of malaria among migrants in Amsterdam-Zuidoost]. AB - In a general practice in Amsterdam Southeast in 1998 a delayed first attack of Plasmodium ovale infection was diagnosed in a 13-year-old girl from Ghana, malaria tropica with a low parasitaemia index in a 43-year-old Ghanaian man and a 8-year-old Ghanaian girl, and Plasmodium vivax infection in a 44-year-old Surinam woman. The Ghanaian patients had visited their native country, the Surinam woman had contracted the infection during a visit to India. All patients responded well to antimalaria medication. These patients were among a total of 6 patients of non Dutch origin diagnosed with malaria in 1998 in this general practice. Four patients had not taken any prophylactic drug and two had not used the drugs properly. A relative increase of malaria in immigrants has been seen in the Netherlands and elsewhere in Europe in recent years. Underestimation of the risks and lack of knowledge of malaria and of the changing epidemiology make people of ethnic minorities travel without taking appropriate precautions. New, creative ways of communication and information will have to be explored to reach these migrant communities. PMID- 10674109 TI - [Cervical smears unsuitable for exclusion of cervical carcinoma]. AB - In women with contact bleeding or vaginal discharge, cervical smears are often made to exclude the possibility of cervical carcinoma. During the past ten years, several women sued pathologists for failure to diagnose cervical cancer because of false-negative results entailing a diagnostic delay of years. This, however, is unjustifiable: clinicians have to consider the aim of performing a test and always have to interpret the outcome within the clinical context. If the indication for testing is to exclude cervical cancer with enough certainty, a false-negative rate possibly as high as 45% makes the smear definitely unsuitable for this aim. PMID- 10674110 TI - [Female circumcision: histories of 3 patients]. PMID- 10674111 TI - [Incidence and risk factors for eye injuries sustained at fairs: metal particles in the eye after a ride of dodgem cars]. PMID- 10674112 TI - [Physical diagnosis--paradoxical pulse]. PMID- 10674113 TI - [Missed infections in immigrant children and the medical care for underage refugees]. PMID- 10674114 TI - [Unusual form of ileus, except in elderly patients?]. AB - Two women aged 88 and 92, recently admitted to hospital, were diagnosed with gallstone ileus. Over a longer period of time they had suffered intermittently from abdominal pain related to a migrating and impacting stone. This disease is mostly diagnosed correctly during exploratory laparotomy for persistent intestinal obstruction. CT scanning, however, is a new and helpful way to early diagnosis. Both women had successful surgery. Gallstone ileus is a disease of the elderly; its early diagnosis is important. CT can be very helpful in this respect. PMID- 10674115 TI - [Dutch research on the effectiveness of medical prescription of heroin; background, research design and preliminary results]. AB - In the Netherlands the total number of heroin addicts amounts to approximately 25,000. Of these about 70% is in contact with the treatment system. The remaining 30% have not been seeking help, believes that no help is needed or has lost faith in a better future altogether. Of those who are in treatment, 30% attempt to stop the use of opiates through participation in drug free abstinence oriented outpatient or inpatient treatment programs. The remaining 70% have given up the outlook of a drug free existence at least temporarily, and they participate in a methadone maintenance program directed at stabilizing drug use, harm minimization and social integration. In two-thirds these goals are not or only partially achieved. For these patients additional treatment options are needed. Medical prescription of heroin is such an option. However, currently no data are available on the effectiveness of this option. The Dutch study on the effectiveness of medical prescription of heroin is an attempt to obtain these data. In the study, simultaneously two randomized trials are being executed: one with inhalable and one with injectable heroin. In these trials, 625 (375 inhalers and 250 injectors) chronic treatment-resistant heroin addicts who are currently enrolled in a methadone maintenance program are offered heroin (in combination with oral methadone) seven days per week, three times per day for a period of six to twelve months. It is a multi-center study with eight treatment units in six cities in the Netherlands (Amsterdam, The Hague, Groningen, Heerlen, Rotterdam, Utrecht). At this moment 180 patients have been randomized. During the treatment no medical complications have been observed and no serious public order or safety problems have occurred. Study participants have been very compliant both with the treatment regimen and the research requirements. The latter is indicated by the fact that 85% of all the two-monthly assessments have been completed. PMID- 10674116 TI - [Therapeutic angiogenesis through intramuscular injection of the gene for vascular endothelial growth factor (VEGF)]. AB - Therapeutic angiogenesis by injection of growth factors or genes encoding for these appears a promising strategy for treatment of critical limb ischaemia but is currently still in an experimental phase. Among the cytokines that promote angiogenesis, i.e. the postnatal growth of blood vessels due to activation and proliferation of endothelial cells, vascular endothelial growth factor (VEGF) takes a special position. VEGF has proven to be a powerful angiogenic factor in vivo that specifically promotes migration and replication of endothelial cells. The specificity of VEGF for endothelial cells, compared with the more widespread effects of other factors such as fibroblast growth factor (FGF), may provide a broader therapeutic potential. The recombinant form of the human VEGF gene can be injected intramuscularly as a plasmid (phVEGF). The angiogenic effect of phVEGF can be explained in several ways: by direct stimulation of proliferation and migration of endothelial cells in vivo; by recruitment of previously existing collaterals; and by improving vasomotor function of vessels in ischaemic tissue. PMID- 10674117 TI - [Non-oral routes of administration of psychotropic agents]. AB - For patients not able to take oral medication, a psychotropic agent may be selected not only according to its therapeutic and side effects but also to the optional alternative routes of administration. Partly because of the various options of routes of administration haloperidol is the first choice 'classic' antipsychotic medication. As to 'atypical' antipsychotics, the preparation of an ill-tasting clozapine liquid is possible. Lorazepam, clorazepate and diazepam injections (oil in water emulsion) or diazepam per rectiole are non-oral anxiolytics of first choice. Clomipramine and amitriptyline are the only parenteral antidepressants available in the Netherlands. Lithium tablets without controlled release may be administered as a suspension via a catheter. Carbamazepine or valproate may serve as a parenteral alternative for lithium. In case of extrapyramidal side effects of antipsychotics, biperiden or promethazine may be administered parenterally. PMID- 10674118 TI - [Poor prognosis of osteosarcoma of pelvis; 39 years of registration by the Netherlands Commission on Bone Tumors]. AB - OBJECTIVE: To evaluate the results of treatment in patients with osteosarcoma of the pelvis. DESIGN: Retrospective. PATIENTS AND METHODS: Sixty-five patients with pelvic osteosarcoma registered in the files of the Netherlands Committee on Bone Tumours over 1957-1995 were reviewed. Complete information was obtained on 62 cases: 27 males and 35 females, median age was 31 years (range: 12-83). Distant metastases were present in 11 patients (stage IIIB; 18%), while 50 patients had stage IIB osteosarcoma (81%) and 1 patient stage IB osteosarcoma (2%). The results of treatment and survival were determined. RESULTS: Median survival was 14 months (2-177), the 5-year survival 15%. Distant metastases developed in 27 of 42 (64%) patients with curatively treated stage IIB osteosarcoma, with prolonged metastasis-free survival after chemotherapy treatment (p < 0.0012). Survival in patients with stage IIB pelvic osteosarcoma was better after surgical resection of the primary tumour than after no operation (p < 0.0054); (neo)adjuvant chemotherapy gave no longer survival than surgery alone (p < 0.083). CONCLUSION: The prognosis of pelvic osteosarcoma was poor, despite modern multimodality treatment regimens, including surgical resection and chemotherapy. PMID- 10674119 TI - [Genetic screening for familial hypercholesterolemia in 1992-1997: primarily younger patients in the care of family physicians]. AB - OBJECTIVE: To estimate the proportion of patients with familial hypercholesterolaemia (FH) who were identified with hypercholesterolaemia in general practice prior to screening by means of pedigree research and DNA analysis by the National Foundation for the Identification of Familial Hypercholesterolemia (StOEH). DESIGN: Retrospective. METHOD: General practice files of FH patients, diagnosed through genetic screening by the StOEH in 1992 1997 whose general practitioner's (GP's) practice in Amsterdam, Haarlem or Alkmaar, were studied for cholesterol and FH related information documented in the period prior to the screening. RESULTS: Out of the 121 persons selected 80 agreed to the study; one GP refused to co-operate. There was no difference between respondents and non respondents with regard to age, sex or domicile of the GP. In 48 of 79 (61%) general practice files studied, cholesterol measurements were reported prior to screening; 39 patients (49%) had hypercholesterolaemia and 29 (37%) were being treated with cholesterol lowering drugs. Mean age of the FH patients who had no record of their cholesterol levels was 25.1 years (SD: 17.0) at the time of screening, 22 years younger than the mean age of FH patients who did have cholesterol levels on record prior to screening (47.1 (SD: 18.4); p < 0.0001). CONCLUSION: Of the FH patients identified through family based genetic screening especially the younger FH patients are newly brought to the attention of their GP. PMID- 10674120 TI - [Gender factors in the selection of training for a medical specialty]. AB - OBJECTIVE: To investigate the choice-making process of male and female doctors regarding their wishes for a career, and their behaviour in the event of their trying to obtain a resident's post. DESIGN: Descriptive. METHODS: An inquiry by telephone was held in 1995 among a group of 600 doctors all graduated from university in 1993 in the Netherlands. Of the respondents (n = 490; 82%) 57% of the female and 63% of the male doctors aspired to become a resident in a hospital. The 293 respondents who wanted to obtain a resident's post were asked for the factors that influenced their choices and to what extent they did. The scores ranked from 1 ('very positively') to 5 ('very negatively'). RESULTS: Within 2 years after graduating from university 26% worked as a resident. In their choice for a specialty male doctors were positively influenced by technology (mean score: 2.9 versus 2.5), status and income (5.9 versus 5.6) and scientific activities (2.4 versus 2.1). Women were more influenced by intensive contact with patients (2.0 versus 1.7), favourable working hours and relatively few shifts (10.9 versus 10.3). Of the women 43% wanted to work part time, as against 14% of the men. While the men preferred an informal approach in looking for a post (38%; formal approach: 29%), women were evenly divided: informal approach 36%, formal approach 36%. PMID- 10674121 TI - [Travel experiences in Central- and Eastern Europe: Bosnia--the land behind the mirror]. AB - Bosnia-Herzegovina is an artificial state created after four years of gruesome war; it is composed of two countries intertwined like the pieces of a jigsaw puzzle. An uneasy truce binds the two halves, the Republika Srpska and the Muslim Croat Federation of Bosnia-Herzegovina together. Under the conditions of the Dayton accord these two have to collaborate and unify their administrative systems under the watching eye of the European Union (EU) and thousands of heavily armed soldiers and policemen. One of these EU-sponsored programmes is the unification of systems of acquisition, registration and distribution of medicines, delegated to the EU reconstruction programme PHARE. Whereas the Serbian half used to buy its drugs from Belgrade, the Muslim-Croatian half was almost fully dependent on international aid. Though some of the local experts have been very helpful, both systems are riddled by corruption and inefficiency and a successful outcome will be little short of a miracle. PMID- 10674122 TI - [Unobserved death of an infant: cot death?]. PMID- 10674123 TI - [Unobserved death of an infant: cot death?]. PMID- 10674124 TI - [Unobserved death of an infant: cot death?]. PMID- 10674125 TI - [Public health care after the Bijlmermeer airplane crash; the aftermath]. PMID- 10674126 TI - [Public health care after the Bijlmermeer airplane crash; the aftermath]. PMID- 10674127 TI - Critical reflections on the evolution in vascular surgery. PMID- 10674128 TI - Behcet's disease. An insight from a vascular surgeon's point of view. AB - Behcet's syndrome is a multisystem inflammatory disease with unknown aetiology, vasculitis being its major pathological feature. It runs an undulating course of exacerbations and remissions with a frequency that usually abates with the passage of time. Following its first description in the medical literature in 1937, many clinical manifestations, including recurrent ulcerations, eye and urogenital lesions, pulmonary and vascular involvement with thrombus and aneurysm formation, arthritis and neurological features were reported. Various studies undertaken in two Medical Faculties of Istanbul University showed that more than 5000 patients have fulfilled three or more International Behcet's Disease Study Group Criteria and thus can be diagnosed as Behcet's Disease. A total of 142 cases with pulmonary, 30 cases with arterial (non-pulmonary) and 10 cases with cardiac involvement were demonstrated since 1978. A group of 174 neuro-Behcet's Disease patients (cerebral venous and vena caval thrombosis as the cause of intracranial hypertension, extracranial vertebral artery dissection) was evaluated. Our collected data showed that surgery with special techniques and in highly selected cases may be useful if used with adjuvant medical therapy, but might be fatal or unsuccessful in the majority. No means of aggressive surgical or interventional therapy has a role in altering the course of the pathology itself when used alone and thus medical treatment is crucial to suppress the exacerbations. PMID- 10674129 TI - Clinical and biological relevance of vein cuff anastomosis. AB - Since a significant number of patients do not have suitable autologous saphenous vein for femorodistal bypass, the search for alternative graft material continues. The most commonly used prosthetic material is Polytetrafluoroethylene, however because of the poor patency of these grafts in the below knee position, a variety of techniques have been tried to improve their patency. A series of studies utilizing venous cuffs at the distal anastomosis have showed improved patency of PTFE grafts. We have reviewed the biological basis for graft failure and the literature for possible mechanical explanations. PMID- 10674130 TI - Laparoscopic management of acute small bowel obstruction. AB - Laparosopy is now a well established tool in abdominal surgery. More often it is used in acute abominal situations. We present our experience with laparoscopy and laparoscopic treatment in patients with acute small bowel obstruction. Although it is technically challenging, in carefully selected patients laparoscopy and laparoscopic treatment is feasible and a valid option for treatment. PMID- 10674131 TI - Early feeding after colorectal surgery. Preliminary results. AB - Thirty three patients were placed under early oral feeding after elective colorectal surgery. There were 15 male and 18 female patients, mean age: 52 years. Nasogastric tube was removed as soon as they were widely awake, or on the morning following the afternoon operations. Oral feeding was resumed 4 hours later, and the first meal consisted in a slight solid meal. There was no postoperative mortality or significant morbidity. Liquid and solid oral intakes were resumed 18 and 24 hours respectively after the operation. Tolerance was perfect in 22 patients (66%), good (slight complaints) in 16%, and was considered as fair or bad in the last six cases. Intestinal transit was observed after a median period of 2 days. Tube insertion rate was 12%. No adverse effect on the anastomoses was noted. Data from the literature confirm that early feeding is tolerable and safe after open colorectal surgery. More patients should be included in this protocol to take benefit of the physiological effects of early oral feeding. PMID- 10674132 TI - Sentinel lymph node mapping in the management of high risk malignant melanoma. AB - In patients with malignant melanoma, the selective biopsy of the first draining lymph node, so-called the sentinel lymph node, allows to identify, with a low morbidity, the patients with nodal metastasis that require radical lymphadenectomy and adjuvant systemic chemotherapy. Herein, we report our initial experience in sentinel lymph node biopsy in 16 patients with malignant melanoma. The sentinel lymph node was localised using preoperative lymphoscintigraphy and injection of dye blue. Intraoperatively, the dissection was guided with a gamma probe and by the recognition of the blue nodes. In the 16 cases the sentinel lymph node was localised. In 50% of the cases, multiple sentinel nodes were demonstrated at lymphoscintigraphy and found during surgery. A limited postoperative morbidity was observed in three cases. Three patients presented nodal metastasis and underwent further radical lymphadenectomy. We conclude that sentinel lymph node mapping is a feasible and reproductive procedure. The preoperative lymphoscintigraphy is essential to identify multiple sentinel nodes and guide surgical dissection. The impact of this approach on the overall survival of patients with high-risk melanoma has still to be demonstrated in studies with a long follow-up. PMID- 10674133 TI - Primary neuroendocrine carcinoma of the liver: difficult diagnosis of a rare neoplasm. AB - Primary neuroendocrine neoplasms of the liver are extremely rare: about 30 cases only have been described in the literature. We report the case of a 42-year-old woman with a ten-year evolution. According to the previously reported cases, primary neuroendocrine carcinoma of the liver is usually multicentric, often mimicking liver metastases. The demonstration of the hepatic origin of a neuroendocrine carcinoma is often arduous. A careful surgical exploration and a prolonged follow-up are mandatory. The treatment of choice is surgical resection when possible. For progressive and unresectable disease, hepatic arterial chemoembolization may be considered. However, the prognosis of liver neuroendocrine tumours is much more favorable than that of hepatocellular carcinoma and progression has to be demonstrated before instauration of potentially harmful therapies. PMID- 10674134 TI - Mucinous cystadenocarcinoma of the appendix. A rare tumour of the right iliac fossa. AB - A case of mucinous cystadenocarcinoma of the appendix is presented. The clinical feature is a painful syndrome of the right iliac fossa. In our observation, the diagnosis was not allowed by preoperative imaging. Appendectomy was initially performed and completed by right hemicolectomy and lymphadenectomy after histological diagnosis of the appendicular malignant tumour was forwarded. The prognosis of this tumour is generally excellent providing early diagnosis and wide enough surgery. PMID- 10674135 TI - Splenic artery aneurysm rupture. AB - This is the report of a case with an atherosclerotic splenic artery aneurysm ruptured into the peritoneal cavity which presented with hypovolaemic shock. The patient underwent successfully emergency laparotomy and splenectomy. A brief review of the the literature follows. PMID- 10674136 TI - Off-pump myocardial revascularization for left main stem disease in a high-risk patient. AB - Off-pump complete myocardial revascularization for three-vessel disease is often limited by the difficulty to approach the obtuse marginal branches. A method of coronary artery bypass grafting without cardiopulmonary bypass used in a high risk patient with left main stem and three-vessel disease is described. PMID- 10674137 TI - Rebound thymic hyperplasia after chemotherapy in a patient treated for pulmonary metastases. AB - A 38-year-old patient presented with an anterior mediastinal mass after chemotherapeutic and surgical treatment for lung metastases from a malignant histiocytoma. Because of the risk for tumour recurrence the thymic mass was resected. Thymic hyperplasia was found on pathological examination. In this case thymic hyperplasia is a rebound phenomenon aflcer chemotherapy. It appears to atrophy during the administration of chemotherapy and regrow afterwards. Surgical resection provides the definitive diagnosis and treatment. PMID- 10674139 TI - Medical treatment of newly diagnosed epilepsy. AB - The first step in the treatment of epilepsy is the confirmation of the diagnosis. A correct diagnosis not only includes the epileptic origin of the event, but also the diagnosis of the seizure type and the epileptic syndrome. The second step is to try to find the aetiology of the seizures. Some papers have shown that the prognosis of epilepsy is better if the seizures are treated earlier, but other papers did not find any difference in the long-term prognosis between patients treated after the first seizure or after several seizures. Therefore, one of the most difficult points, after confirmation of the diagnosis, with a first or few seizures will be to identify the risks of relapse in some patients and to immediately treat them and to avoid treating the others who will have only one or rare seizures during their lives without any damage. In most cases, the first treatment will be the prescription of an antiepileptic drug (AED) in monotherapy. If the cause is treatable, it will be treated concomitantly. In generalised epilepsies, especially in idiopathic syndromes, valproate will be the first choice, most of the classic AEDs may worsen some seizure types in these syndromes. In partial epilepsies, there are no statistically significant differences in efficacy between the 4 classic major AEDs (carbamazepine, phenytoin, phenobarbitone and valproate) in pooled data. The choice of the drug should be more influenced by considerations of safety profile, dosing frequency, and costs for equivalent advantages. Accordingly, valproate is a good first choice in patients in whom the epilepsy syndrome is not clearly defined. Efficacy of newer AEDs is similar to old AEDs but most are better tolerated. However, some studies including seizure control, side effects, medical consultation, inpatient, accidental injuries, and laboratory investigations showed that newer AEDs are more expensive in newly diagnosed patients, compared to classic major AEDs and this notion should be taken into account for the prescription. PMID- 10674140 TI - The spectrum of the new antiepileptic drugs. AB - Over the last decade, many new drugs have been added to the therapeutic armamentarium for epilepsy. These drugs differ considerably in their mechanisms of action and, consequently, in their spectrum of efficacy against various seizure types. Oxcarbazepine, gabapentin, tiagabine and vigabatrin are especially useful in the management of partial seizures (with or without secondary generalization) and, probably, also primarily generalized tonic-clonic seizures, with vigabatrin being of particular value also in the treatment of infantile spasms. The spectrum of efficacy of lamotrigine and topiramate is broader than that of the other drugs and includes, in addition to partial and tonic-clonic seizures, also drop attacks associated with the Lennox-Gastaut syndrome. Lamotrigine is also effective against absence seizures, while the activity of topiramate as a potential anti-absence drug has not been adequately explored. Oxcarbazepine, vigabatrin and tiagabine may aggravate myoclonic and absence seizures and, likewise, gabapentin may aggravate myoclonic seizures. Therefore, the latter drugs should not be used (or used with great caution) in patients with syndromes associated with these seizure types. Apart from differences in spectrum of efficacy, side effect profiles also differ considerably from one drug to another, with the risk of serious adverse effects limiting considerably the use of felbamate and vigabatrin. When added to preexisting therapy in patients with refractory epilepsies, the new drugs improve seizure frequency in 15% to 40% of cases, but only rarely freedom from seizures is achieved. In newly diagnosed patients, the efficacy of the new drugs is similar to that of older agents, but further studies are required to confirm the claim that the tolerability of some of these agents is superior to that of established drugs such as carbamazepine or valproate. The new antiepileptic drugs represent a useful addition to the therapeutic armamentarium, but because of limited clinical experience and cost considerations their firstline use cannot be recommended in most situations. PMID- 10674141 TI - Pharmacoeconomic evaluation of medical treatment of epilepsy: where do we stand? AB - Qualitative and quantitative improvements of pharmaco-economic evaluation of antiepileptic drugs have been realized during the last decade. Assessment of medical treatments is mainly performed through cost-minimization studies, owing to similar effectiveness of AED. Studies recently published generally consider hypothetical cohorts of patients and assess resource utilization and medical effects on the basis of clinical trials and expert panels, limiting evaluation to direct costs. But they differ in numerous ways due to: the type of patients and treatment considered, the time span of evaluation, and cost measurement. Comparison between studies is therefore tricky but it seems that, when considering monotherapy treatments, carbamazepine and phenytoin are somewhat cheaper than valproate while lamotrigine is much more expensive but no more effective. However, these and other antiepileptic agents appear as close substitutes when prescribed as adjunctive treatments. Refined tools have been developed to take into account slight differences exhibited on certain outcomes. One direction currently under evaluation relates to quality of life, and this will probably leads to an increasing number of cost-utility studies. Some advances, regarding information on medical resources consumed and outcomes associated with alternative drugs, as well as methodological options, are still needed to fully develop pharmacoeconomic evaluation of antiepileptic treatments. PMID- 10674138 TI - Animal models of focal epilepsy. AB - Animal models are important in the study of the development and expression of focal seizures as well as in the preclinical evaluation of antiepileptic treatment. Many different models are available, including acute and chronic models for simple partial seizures and models for complex partial seizures. Work on models has revealed that the pathophysiology of seizure disorders includes several neurotransmitter and membrane channel alterations. In addition, epileptogenesis of focal epilepsy has been shown to involve the selective loss of neurons and axonal reorganization. Antiepileptic treatment still hinges on three general themes: modulation of voltage-dependent ion channels involved in spike propagation and burst generation, enhancement of GABA-mediated inhibition, and suppression of excitatory amino acidergic activity. Many antiepileptic drugs have proven efficacy against focal seizures in animal models as well as patients. More recently developed antiepileptic drugs may prove to be superior in the alleviation of intractable partial seizures. The three general themes of antiepileptic drug action still dominate the development of antiepileptic treatment strategies. Too much emphasis on the classical models of focal epilepsy may hamper the development of innovative strategies. On the other hand, continued research on new and existing models may broaden our knowledge of the pathophysiological processes underlying focal epilepsy, and inspire new avenues in antiepileptic treatment development. PMID- 10674143 TI - Epilepsy surgery in Belgium, the experience in Gent. AB - Between January 1992 and July 1998, 320 patients were presurgically evaluated for medically refractory epilepsy at the University Hospital of Gent. All patients underwent a comprehensive presurgical evaluation, including extensive neurological history and examination, video-EEG monitoring of interictal EEG and habitual seizures, and optimum magnetic resonance (MR). In a large subgroup of these patients, a comprehensive neuropsychological examination and interictal 18FDG-PET were performed. Subsequently, a bilateral carotid angiography and intracarotid amytal procedure (Wada-test) were planned in 49 patients to establish hemispheric language dominance and bilateral memory function. After proper selection, 23 patients underwent invasive video-EEG monitoring with intracranial implantation of parenchymal and/or subdural electrodes to further document the area of seizure onset. From the initial group of 320 potential surgical candidates, 75 patients (42 males, 33 females) with mean age of 29 years (range: 2 months-55 years) and mean duration of uncontrolled seizures of 15 years (range: 2 weeks-38 years) eventually underwent a surgical procedure. Sixty of 75 patients were on high dose antiepileptic polytherapy. Optimum MR detected structural abnormalities, confined to a limited brain area, in 71 patients. These abnormalities were of space-occupying nature in 31 cases; an atrophic lesion was suspected in 39 patients; a combination of space-occupying and atrophic lesion was seen in 1 case. Structural abnormalities were most frequently located in the temporal lobe (n = 53) and the frontal lobe (n = 10). Video-EEG monitoring documented complex partial seizures in 67 patients with occasional secondary generalisation in 32. Most patients had complex partial seizures of temporal lobe as defined by clinical and EEG criteria. Two patients had only simple partial seizures. Ultimately, an area of seizure onset could be determined in all patients. Temporal lobectomy with hippocampectomy was the most commonly performed procedure (n = 42). In 13 patients, complete lesionectomies were performed for epileptogenic structural lesions in and outside the temporal lobe. In 2 patients, only partial lesionectomies were possible; in 5 patients, only biopsies in combination with partial lesionectomies could be performed. Anterior 2/3 callosotomy was performed in 4 patients and hemispherectomy was performed in 2 patients. Postsurgical seizure control, after average follow-up of 50 months (range: 12-98 months), was excellent in 49 patients who became seizure-free. In these patients, antiepileptic therapy was tapered 2 years after surgery. Patients in whom only biopsies or partial lesionectomies were performed have poor seizure control. Epilepsy surgery is a rewarding therapeutic alternative for patients with medically refractory epilepsy. Comprehensive presurgical evaluation and epilepsy surgery provide excellent neurological, neurophysiological, neuropsychological and imaging research opportunities. PMID- 10674142 TI - Interictal and ictal video-EEG monitoring. AB - PURPOSE: The purpose of this paper is to demonstrate the diagnostic efficacy and therapeutic relevance of video-EEG monitoring in an large patient population with long-term follow-up. PATIENTS AND METHODS: Between October 1990 and May 1997, 400 patients were monitored at the Epilepsy Monitoring Unit (EMU) of the University Hospital in Gent. In all patients, the following parameters were retrospectively examined: reason for referral, tentative diagnosis, prescribed antiepileptic drugs (AEDs), seizure frequency, number of admission days, number of recorded seizures, ictal and interictal EEG, clinical and electroencephalographic diagnosis following the monitoring session. During follow-up visits at the Epilepsy Clinic, we prospectively collected data on different types of treatment and post-monitoring seizure control. RESULTS: 255/400 (64%) patients were referred for refractory epilepsy. 145/400 (36%) patients were evaluated for attacks of uncertain origin. Mean follow-up, available in 225 patients, was 28 months (range: 6-80 months). Mean duration of a single monitoring session was 4 days (range: 2-7 days). Prolonged interictal EEG was recorded in all patients and ictal EEG in 258 (65%) patients. Following the monitoring session, the diagnosis of epilepsy was confirmed in 217 patients. Pseudoseizures were diagnosed in 31 patients (8%). AEDs were started in 19 patients, stopped in 6 and left unchanged in 110. The type and/or number of AEDs was changed in 111 patients. Sixty patients underwent epilepsy surgery. In 48 surgery patients, follow-up data were available, 29 of whom became seizure-free, and 16 of whom experienced a greater than 90% seizure reduction. Vagus nerve stimulation was performed in 11 patients, 2 became seizure-free, and 7 improved markedly. Of the non-invasively treated patients in whom follow-up was available (n = 135), 70 became seizure-free or experienced a greater than 50% reduction in seizure frequency; 51 patients experienced no change in seizure frequency. Outcome was unrelated to the availability of ictal video-EEG recording. In patients with complex partial seizures, seizure control was significantly improved when a well-defined ictal onset zone could be defined during video-EEG monitoring. CONCLUSION: Prolonged interictal EEG monitoring is mandatory in the successful management of patients with refractory epilepsy. Ictal video-EEG monitoring is very helpful but not indispensable, except in patients enrolled for presurgical evaluation or suspected of having pseudoseizures. PMID- 10674144 TI - Cost-benefit of epilepsy surgery. AB - A cost-benefit study of Epilepsy Surgery (ES) evaluates the monetary consequences of the societal gain by reduced production loss. High direct cost of ES has been addressed by suggesting simpler diagnostics, preference for certain investigations, and weighing the direct cost against the postoperative savings. New MRI technology has simplified diagnostics and, currently, the surgical procedure is more accurate. A shortlasting procedure will reduce theater cost and hospitalization. Resective procedures achieve freedom from seizures in 45-90%. Postoperative worsening may be burdened by permanent morbidity, or may cause death. Re-operation, an extra direct cost, yields freedom from seizures in 45 50%, being thereby justified health economically. If ES stops seizures at the expense of impaired cognition, it is of limited help in restoring quality of life. In adults, surgery often is performed late, and the social status may have declined for the patient. Children with epilepsy and their families experience social restrictions difficult to normalize. A successful outcome unaccompanied by other improvement in life may turn into disappointment. Operated patients employed preoperatively continue to be so postoperatively, or maintain their employment which may also improve, an effect experienced even after a re operation. But, a limited gain in employment after surgery has likewise been reported. Early surgery has a positive effect on education and occupation. The postoperative income of adults may increase significantly. Similarly, successful pediatric surgery is cost-effective for the child and the family. Using seizure free rate as effectiveness measure shows that ES is cheaper than AED treatment alone. The high cost for advanced diagnostics should be seen in a long-term perspective. The reduction in life long indirect cost after successful pediatric surgery has been evaluated. PMID- 10674146 TI - Presurgical assessment and surgical treatment for epilepsy. AB - In the last ten years, dramatic advances in surgical treatment options and techniques have allowed surgical intervention for patients who would otherwise not have been considered as surgical candidates. In this article, a multidisciplinary, logical decision algorithm for a rational approach to surgical treatments is outlined. A carefully considered hierarchy is presented that provides for maximized seizure improvement outcomes. Topics presented include temporal lobectomy, detailed discussion of dominant temporal lobectomy and speech sparing techniques, neocortical resection, the use of subdural electrode array, depth electrodes, and strip electrodes, multiple subpinal transection, vagus nerve stimulation, and corpus callosotomy. The application of these various techniques to maximize surgical outcome are discussed. PMID- 10674145 TI - Cost-benefit of vagus nerve stimulation for refractory epilepsy. AB - PURPOSE: Vagus nerve stimulation (VNS) is an established treatment for patients with medically refractory epilepsy who are unsuitable candidates for conventional epilepsy surgery. VNS requires an initial financial investment but apart from our own previous study there are no reports on cost-benefit published to date. The purpose of this paper is to assess prospectively the cost-benefit ratio of VNS in a series of patients with long term follow-up. METHODS: Our experience with VNS comprises 25 patients of whom 20 with sufficient follow-up will be further discussed. These 20 patients have a mean post-implantation follow-up of 26 months (range: 6-50 months). Mean age was 30 years (range: 12-45 years); mean duration of epilepsy 17 years (range: 5-35 years). We prospectively assessed seizure frequency, prescribed AEDs, number of hospital admission days and side effects and calculated the epilepsy related direct medical cost and compared this with pre-implantation data. RESULTS: Mean seizure frequency decreased from 14 seizures/month (range: 2-40) to 9 seizures/month (range: 0-30) (p = 0.0003). The mean yearly epilepsy related direct medical costs per patient dropped from 6,682 USD (range: 829-21,888 USD) to 3,635 USD (range: 684-12,486 USD) (p = 0.0046). The mean number of hospital admission days was reduced from 16 days/year (range: 0-60) to 4 days/year (range: 0-30) (p = 0.0029). CONCLUSION: VNS is an efficacious and cost-beneficial treatment for refractory partial seizures. PMID- 10674147 TI - A psychoanalysis of love: the patient with deadly longing. AB - Freud made some seriously mistaken assumptions about mature love based on his notion about the role unconscious guilt plays in human affairs. Clinical data are presented here about a young man, who implicated in the suicides of two of his colleagues, was more concerned with the failure of his love affair in college than with guilt for his involvement in the suicides. By tracing the functions of shame--a concept the author contends that Freud failed to understand properly--in the patient's relationship with his parents, the author formulates a proactive and optimistic theory of mature love to replace Freud's pessimistic and reductionistic explanation. PMID- 10674148 TI - Interpreting transference in the supervision of psychoanalytic psychotherapy. AB - Supervisors of analytic psychotherapy have long wrestled with the question of whether interpretation has a legitimate role in dealing with supervisee countertransference and the transferences of the supervisory experience, itself. Currently, the majority view relies on didactic methods to deal with these transferences and avoids interpretation as incompatible with, even dangerous to, the supervisory task. This paper takes issue with this view and uses a clinical example to illustrate the impact and irreplaceable value of direct interpretation in supervision. It demonstrates that interpretation of a resistance in the supervisee can fundamentally and beneficially affect the therapeutic relationship, the supervisory process, and can have unanticipated diagnostic significance. It also demonstrates that, when applied with the same appropriateness and tact taken for granted in psychotherapy, concerns that supervisory interpretation will be traumatizing or counterproductive are unwarranted. PMID- 10674149 TI - Therapy manuals and the dilemma of dynamically oriented therapists and researchers. AB - The presence of therapy manuals in clinical settings is increasing and the related concepts of adherence and competence are becoming familiar. The benefits of manuals for research and training are evident. However, negative clinical effects have also been reported. Dynamically oriented clinicians and researchers who use manuals face a dilemma. Although there is a need to control aspects of technique for research and training purposes, there is also a need not to control the process of therapy because unpredictability is an intended part of the process. Issues associated with the dilemma are reviewed and a possible solution is provided that has proven helpful in an active clinic and research setting. It involves the use of manuals that emphasize general guidelines rather than detailed technical behaviors. PMID- 10674150 TI - Performative statements and the will: mechanisms of psychotherapeutic change. AB - Performative statements are much discussed in the philosophy of language, e.g., by the linguistic philosopher, J. L. Austin, where they are distinguished from descriptive statements. Whereas the latter merely describe a current state of affairs from an external standpoint, performative statements enact a new state of affairs merely by being spoken, as when a minister pronounces a couple married during a wedding ceremony. Performative statements are words and deeds at the same time. They are special kinds of statements, requiring certain unique circumstances and relationships so that they can function validly. Leston Havens has made the connection between the ability to make performative statements and the setting of psychotherapy. He has asserted that performative statements are an important part of the psychotherapeutic repertoire and may be an important force in bringing about psychotherapeutic change. In this paper, I try to locate the mechanism by which performative statements may achieve this effect, suggesting that this occurs by means of influencing the patient's will directly. Performative statements work on the will, much as descriptive statements influence the intellect. Philosophical ideas about the will, like Aristotle's notion of weakness of will, may explain some of the phenomena of psychotherapy, including resistance. PMID- 10674151 TI - "Generative caring" psychotherapy for patients who are reluctant to talk. AB - Patients seek psychotherapy for relief of symptoms and resolution of problems in living. Yet, they sometimes balk at the prospect of having to choose a topic without being prompted by therapist questions. This paper suggests that this apparently self-sabotaging behavior represents an acting out of a fundamental existential dilemma for many individuals, i.e., to be or not to be an adult. Adulthood comes with the heavy burden of responsibility for making life choices, and the possibility of making the wrong choices. The psychotherapeutic situation, with its demand that the patient choose a topic, is a microcosm of the adult adult relationship. The author proposes that the proper treatment for these patients is Generative Caring Psychotherapy, which promotes emotional growth and facilitates patients' transition from childhood dependence to adult responsibility. PMID- 10674152 TI - A time-saving technique for the treatment of simple phobias. AB - Two cases are presented in detail and two in summary fashion to illustrate a technique that can frequently be used instead of systematic desensitization to reduce the time needed to treat simple phobias. This method combines techniques developed by Strategic Therapy and Critical Incident Debriefing. Symptoms that the patient experiences as out of control are prescribed by the therapist and then normalized. For example, a 26-year-old woman with a fear of social situations learns that it is not unusual for people to feel somewhat awkward and anxious as they try to reestablish themselves with friends after being away from them for a long period of time. Therapy taught her to accept rather than fight her initial anxiety in these situation. Another client with claustrophobia was taught to imagine himself getting anxious and telling himself, "yes, this is exactly what I expect. I am going to get anxious, and my anxiety will increase, but it isn't going to get higher than a '5' (on a 1-10 scale), and I can handle that." When these interventions are successful, the anxiety initially experienced in a phobic situation as a signal for panic is reinterpreted in new situations as expectable. This reframing renders the anxiety manageable. The treatment of two additional patients is briefly presented to further illustrate the application of this approach. Their phobias included a fear of sweating in public and a fear of sleep. PMID- 10674153 TI - Short-term dynamic therapy as a unique container. PMID- 10674154 TI - The evolution of the internal dialogue during the psychoanalytic psychotherapy process. AB - This article illustrates curative changes occurring in psychoanalytic psychotherapy by developing the internal dialogue in the mind. By the internal dialogue, I mean the internalization of the therapy process during the recurrent therapy sessions so that the external dialogue between therapist and patient becomes the corresponding internal structure of the patient. In this way, there is a development of the patient's capacity to identify himself and the therapist as separate, sentient, thinking, and reflecting individuals, who have a free internal world of their own. The evolution of the internal dialogue takes place by gradually progressing symbolization achieved through a four-step symbolization reflectiveness approach. This process is one of the specific curative factors in the psychotherapeutic treatment of borderline patients. The psychodynamics are illustrated by material from one session each of the early and the final stages of therapy, showing a shift from monologue to internal dialogue. The evolved internal dialogue is a central part of the patient's budding thinking capacity, which creates a ground for her subjectivity and autonomy. This study underlines and specifies three factors as cornerstones of the evolution of identity in psychotherapy: symbolization, reflectiveness, and the internal dialogue. PMID- 10674155 TI - Coming of age: the adolescent in psychotherapy. AB - Psychotherapy clients in their adolescence are receptive to self-exploration. When given the opportunity to reduce external distractions and directives, these young individuals are serious seekers of emotional well-being. They also seek quality and genuine interpersonal relationships. Gentle guidance within a safe container encourages adolescents to explore their inner world. PMID- 10674156 TI - Psychoanalytic pluralism and its vicissitudes. PMID- 10674157 TI - A verified case of recovered memories of sexual abuse. PMID- 10674158 TI - A survey of fragile X syndrome in a sample from Spanish Basque country. AB - Fragile X syndrome is the most common inherited form of mental retardation. The syndrome is associated with a CGG repeat expansion in the 5'UTR of the first exon of the FMR1 gene. This gene maps to Xq27.3 and coincides with the cytogenetic fragile site (FRAXA). The present study deals with the prevalence of fragile X syndrome among individuals with mental retardation of unknown cause from institutions and special schools from the Spanish Basque Country. Results of cytogenetic and molecular studies, performed in a group of 134 unrelated individuals (92 males and 42 females) are presented. The cytogenetic marker at Xq27.3 was identified in 12 patients. Other chromosomal abnormalities were found in two cases that this and previous studies confirmed as Angelman and Prader Willi syndromes. Two males, in whom the cytogenetic marker was identified, were found negative for FRAXA and FRAXE expansion at the molecular level. The present study shows that the frequency of the FRAXA full mutation in individuals of Spanish non-Basque origin is in the range of other Spanish populations. In the sample of Spanish Basque origin we have not found cytogenetic FRAXA site expression, and the CGG repeat size of FMR1 gene is in the normal range. The significance of these results are discussed. PMID- 10674159 TI - Double telomeric signals on single chromatids revealed by FISH and PRINS. AB - FISH probes for all human telomeres and specific telomeric probes that hybridize to unique sequences on individual chromosomes have been used to characterize the telomeric hybridization pattern of human peripheral blood lymphocytes and bone marrow cells in interphase and metaphase chromosomes. We have identified the existence of double hybridization signals on chromatids both with the (TTAGGG)n telomere repeat arrays and on non chromosome-specific subtelomeric regions as well as on chromosome-specific sequences located several kilobases from the end of chromosomes. Preliminary results using cosmid or YAC probes that hybridize to regions rich in GC sequences also revealed double fluorescent spots on a single chromatid. Double spots were detected by PRINS on terminal and interstitial telomeric sequences in avian cells. The significance of this phenomenon is discussed based on some models of chromatid and DNA organization such as uninemy, looped chromatid organization and quartet DNA structures. The occurrence of double spots should be taken into consideration for the clinical cytogenetic diagnosis of duplications. PMID- 10674160 TI - Cerebellar dysgenesis and mental retardation associated with a complex chromosome rearrangement. AB - Cerebellar hypoplasia, mild mental retardation, skeletal abnormalities, and ataxia were present in a 40 years old patient with a complex chromosome rearrangement (CCR). Chromosomes 2, 5, 16, and 17 were involved in the CCR. For the definition of the eight breakpoints leading to the rearrangement FISH with whole chromosomes paintings and specific telomeric probes was employed. Gene disruption, positional effect variegation, and sub-microscopic deletions are all possible causes for the abnormal phenotype observed in the patient. PMID- 10674161 TI - Proximal trisomy 13q and distal monosomy 8p in a dysmorphic and mentally retarded patient with an isodicentric chromosome 13q and a 13q/8p translocation chromosome. AB - A 40 year-old dysmorphic and mentally retarded female is reported with a de novo unbalanced chromosomal rearrangement (karyotype: 46,XX,der(8)t(8;13)(p23;q123),idic(13)(pter-->q123: q123-->pter) resulting in an isodicentric chromosome 13 and a double aneusomy including partial trisomy 13 (13pter-q123) and distal monosomy 8p (8pter-p23). The main clinical findings consist of developmental/mental retardation, behavioural disturbances and minor congenital defects, not consistent with the clinical pattern of either of the two aneusomies. A mechanism for the chromosome rearrangement is proposed and the absence of specific physical findings in the present patient is discussed in the light of the available literature data. PMID- 10674162 TI - Novel translocation t(3;11)(p21;q24) in multiple myeloma characterised by FISH. AB - We present a 72 year old man with multiple myeloma (MM). Cytogenetic and FISH analysis of bone marrow aspirate showed a novel translocation der(11)t(3;11)(p21;q24). The unbalanced karyotype led to substantial partial trisomy for chromosome 3p and small partial monosomy 11q. Structural rearrangements of chromosome 3 are uncommon in MM and these are reviewed. The patient died 2 years after the diagnosis of MM was made. PMID- 10674163 TI - Schinzel-Giedion syndrome with severe deafness and neurodegenerative process. AB - A case of Schinzel-Giedion syndrome with a follow-up of two and a half years is reported. In addition to the classical features of the syndrome, the patient had severe hearing loss with ossicular and cochlear malformations, alacrymia, and progressive neurodegenerative disease. PMID- 10674164 TI - A novel and very peculiar HincII polymorphism in the 5' region of the human neurofibromatosis type 1 (NF1) gene. AB - We report a HincII polymorphism in the 5' end of the neurofibromatosis type 1 gene (NF1) as detected with a probe made of exons 1 to 4a (nucleotides 2 to 401 of the cDNA). This HincII site is most probably in an intron. Evidence presented suggests the probe reveals not one but two similar polymorphisms. PMID- 10674165 TI - [French Society for Human Genetics. "Genetics in Practice" Commission. Core scientific data of use in genetic counseling. Hemochromatosis]. PMID- 10674166 TI - [French Society for Human Genetics. "Genetics in Practice" Commission. Core scientific data of use in genetic counseling. Familial Mediterranean fever]. PMID- 10674167 TI - Influence of dietary phosphorus depletion on central pathways of intermediary metabolism in rats. AB - Studies on P depleted rats compared with control animals kept under pair fed conditions were carried out. Dietary P depletion led to reduced weight gain and a decreased food conversion ratio in comparison with pair fed control animals. Higher N retention, higher urea concentrations in plasma, liver and kidney tissues and a significant reduction of renal glutamate dehydrogenase--a central enzyme of amino acid degradation in kidney mitochondria--in P depleted animals indicated changes in N utilization and N excretion. While lipid metabolism was not affected by P depletion, carbohydrate metabolism was substantially changed in the kidney: Activity of fructose diphosphatase was significantly reduced, implicating a reduced gluconeogenesis in P depletion. Possible mechanisms of these metabolic effects in P depletion are discussed. PMID- 10674168 TI - Whole body protein turnover of growing rats in response to different dietary proteins--soy protein or casein. AB - Whole body protein turnover was studied in growing rats fed restrictively on isoenergetic (GE 17.6 MJ/kg DM) and isonitrogenous (104 g CP/kg DM) diets based on soy protein isolate or casein supplemented with D,L-methionine. During each of the three separate experiments six male Fischer rats per group were housed individually in metabolic cages at 24 degrees C. Prefeeding of both dietary groups up to similar body weights at the start of the main experimental periods (105-134 g) lasted up to 16 d for casein-fed rats and up to 30 d for the soy protein-fed rats. Following the energy and nitrogen balance periods whole-body protein synthesis was estimated by the end-product method using a single tracer dose of a mixture of 15N-labelled amino acids. Fractional protein accretion rate [% of the protein pool accreted per day] was significantly lower in soy protein rats than in casein-fed rats in all three experiments whereas fractional synthesis rate was not significantly lower. Therefore, protein breakdown subsequently calculated as the difference between synthesis and accretion showed a tendency towards higher values in this group. In soy protein-fed rats also a tendency towards higher excretion of 3-methylhistidine as a marker of myofibrillar protein breakdown was observed. It is concluded that increase in lean tissue growth resulting from improved protein quality is brought about by changes of both rates, by small increase of protein synthesis and by reduced rate of body protein breakdown. PMID- 10674169 TI - The course of phosphorus excretion in growing pigs fed continuously increasing phosphorus concentrations after a phosphorus depletion. AB - A balance study was performed in order to quantify the effect of continuously increased phosphorus (P) intake on faecal and urinary P excretion. The aim was to quantify the level of intake where regulatory P excretion becomes relevant for comparative digestibility measurements on P, and when the pig adapts its urinary P excretion to increased P intake. Phosphorus intake of growing pigs was continuously increased on a daily basis starting at a marginal level and P excretion via faeces and urine was continuously followed for 92 days. Two semi synthetic diets were prepared with different proportions of Na2HPO4 resulting in 2.4 (diet 1) and 6.3 (diet 2) g P/kg DM. Concentration of Ca was adapted to achieve a Ca supply approximately 3.1 fold the digestible P supply. Six castrated male crossbred pigs (31 kg BW) were kept individually in metabolism crates after they had undergone a 14 d P depletion period during which they were fed diet 1 solely. Pigs received 1.04kg of diet 1 per day throughout the experiment, and each day the amount of feed and P supplied to pigs from diet 2 was increased by 12 g and 69 mg, respectively. ME supply was approximately 2.4 fold maintenance and average daily BW gain of pigs during the entire experiment was 690 +/- 30 g. While intake increased linearly, faecal excretion of P and Ca increased non linearly and could be best described by third order polynomial functions. The proportion of ingested P not excreted via faeces followed a quadratic type of curve with a maximum of 81% at 25 days on experiment and P intake of 4.0 g/d. Thereafter, the proportion decreased continuously. The digestibility of P from diet 2, determined by the slope ratio technique, was constant and not affected by P intake up to a P intake of 5 g/d. Renal P excretion did not exceed inevitable losses until day 60 and increased exponentially thereafter when body P reserves were restored. It is concluded, that an adaptation to surplus P supply occurred earlier on the intestinal than on the renal level. While faecal P excretion appeared regulated depending on the actual requirement for P retention, the regulation via urine depended on the P status of the pig. Once the renal P excretion of growing pigs exceeds a level of 25 mg/d, intake of digestible P cannot be regarded sufficiently low to measure P digestibility as a capacity of the feedstuff. PMID- 10674170 TI - Passage of ribonucleic acid along the intestine of sheep. AB - Sheep (Flemish female x Texel male, 55 kg BW), fitted with a PVC cannula in the dorsal rumen and single T-shaped PVC cannulas in the proximal duodenum, distal duodenum, mid-jejunum and terminal ileum were fed hay or hay-concentrate diets at various levels of nitrogen and cell walls (NDF) (22 to 32 g N/d; 150 to 699 g NDF/d). Co-EDTA and Cr-NDF were used as markers to measure the flow rate of digesta. Ribonucleic acid (RNA) intestinal digesta and in rumen bacteria was determined with orcinol after extraction with sodium chloride, precipitation with tungstophosphoric acid and alkaline hydrolysis. The RNA:total N ratio in bacteria, harvested from the rumen, amounted to 0.70 (CV 4.4%). The apparent digestibility of RNA in different sections of the intestine was higher than of total N. About 6% of RNA entering the duodenum disappeared between the proximal and distal duodenum. At jejunum, the net disappearance of RNA amounted to 68% of the quantity which entered the proximal duodenum. A higher result of 71% was obtained at the ileum. Total net disappearance of RNA between the proximal duodenum and rectum averaged 75%. Sixteen percent of RNA leaving the ileum was apparently digested in the large intestine. The true digestibility of RNA between the proximal duodenum and the terminal ileum, as estimated by multiple regression analysis, amounted to 78%. Of the amount of RNA entering the ileum, 24% was of endogenous origin. At ileum, the RNA passage was positively related to the ileal flow of NDF (R2 = 0.67) and N (R2 = 0.94). The passage of RNA increased by 3 mg RNA per g ileal indigestible NDF. Ileal endogenous N consisting of approximately 2% of endogenous RNA-N. In conclusion, the digestion capacity in the first part of the small intestine is high. Rising flows of indigestible cell walls and nitrogen increase the loss of ileal RNA. Further, using RNA as a microbial marker to assess the amount of microbial protein entering the duodenum of ruminants, digesta samples should be collected immediately post pylorus at the proximal duodenum, in order to avoid underestimation of the microbial protein synthesis in the rumen. PMID- 10674171 TI - Morphological and functional development of the rumen in the calf: influence of the time of weaning. 1. Morphological development of rumen mucosa. AB - The objective of this study was to determine whether the nutritional regimen of rearing calves would influence the morphometric and histological development of rumen mucosa. Twelve male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I), early weaned (6 weeks) group (II) and late weaned (9 weeks) group (III). All animals received additional solid feed. Animals of group I were slaughtered after 6 weeks of age, whereas those in groups II and III were slaughtered after 9 weeks of age. At slaughter, the ruminal digesta amounted to 2035 g (milk group), 3092 g (late weaned group) and 5374 g (early weaned group). The differences in the ruminal molar percentage of SCFA were not significant. There was a trend for lower pH and higher SCFA concentrations in the order late weaned, early weaned and milk fed animals (pH: 6.4, 6.6 and 6.7, respectively; SCFA: 96, 87 and 77 mmol/l, respectively). The mean length (1.07 mm in milk group, 1.45 mm in late weaned group and 1.87 mm in early weaned group), width (0.43, 0.58 and 0.71 mm, respectively) and surface of papillae (190, 232 and 241 mm2/cm2 mucosa, respectively) increased with both the age of the animals and the elevated intake of solid feed, whereas the number of papillae (210, 140 and 92 per cm2 mucosa, respectively) decreased. In both milk fed groups type A and B corneal cells were present in the Stratum corneum, whereas in the earlier weaned calves type C-cells could be also seen. These findings indicate a more advanced stage of development of the rumen epithelium in the earlier weaned calves fed higher amounts of concentrate and hay. PMID- 10674172 TI - Calculation of utilizable crude protein at the duodenum of cattle by two different approaches. AB - The duodenal flow of utilizable crude protein (crude protein minus endogenous protein) in cows was estimated using dietary parameters, first by multiple regression and secondly by the addition of microbial protein and undegraded feed protein. These estimates were compared with 327 results from experiments conducted with fistulated cows in Braunschweig-Volkenrode and Rostock Dummerstorf. The regressions and the measurements for microbial protein synthesis as well as feed protein degradation and organic matter fermentation in the rumen were based on the same experimental data set. The prediction of utilizable crude protein (uCP) at the duodenum by regression with digested organic matter (kg DOM) and undegraded feed protein (g UDP) as predicting variables, was more accurate than the value given by microbial protein synthesis and rumen protein degradability. The regression model [g uCP = [188.5-(116.5 (UDP/CP))] DOM + 1.03 UDP] had the highest coefficient of determination (r2 = 0.91) and the lowest coefficient of variation (cv = 8.6); indicating the model's superiority over the other method of estimation. PMID- 10674174 TI - Effect of two variables on the fatigue performance of acrylic bone cement: mixing method and viscosity. AB - The goal of the present work was to establish the relative influence of one exogenous variable versus one endogenous variable on the fully-reversed tension compression fatigue performance of bone cement. The method used to mix the cement constituents was the exogenous variable, while the viscosity of the mixed cement dough was the endogenous variable. Two commercial cement formulations (Palacos R and Osteopal) and two cement mixing methods (hand mixing and vacuum mixing) were used. It was found that for a given mixing method, cement viscosity exerts a marginal influence on fatigue performance. On the other hand, for a given cement formulation, vacuum mixing led to a statistically significant improvement in fatigue performance. The present results demonstrate the superior influence of mixing method over cement viscosity. PMID- 10674173 TI - [The effect of palm oil and safflower oil in the feed of parent fattening hens on fertility, hatchability and growth of progeny]. AB - The aim of two experiments with broiler breeder hens was to evaluate the effect of diets containing palm butter or safflower oil (25 g and 50 g/kg feed, resp.) on fertility, hatchability and growth of progeny. Especially the incorporation of oleic and linoleic acid in egg yolk reflected the dietary fatty acid source. Eggs were collected and stored in the incubator at a hen age of 31, 40, 50, and 60 weeks. Hatched chicks were reared over 5 weeks. The number of fertile eggs (Experiment 1 and 2, 75 and 88%, resp.) differed between the experiments (P < or = 0.05). Neither embryonic mortality nor hatchability (Experiment 1 and 2, 76 and 78%, resp.) were significantly affected by fatty acid composition of yolk. No clear maternal dietary effect was recorded on chicken weight at hatching (Experiment 1 and 2, 43.3 g and 43.7 g, resp.) and at 35 days of age (Experimental 1 and 2, 1676 g and 1764 g, resp.) The fatty acid composition in the analysed egg yolk sac of chicks showed a different fatty level but corresponded to fatty acid composition of breeding eggs before incubation. According to a decreased level of docosahexaenoic acid in egg yolk due to increased incorporation of linoleic acid, the content of this fatty acid was also diminished in phospholipids of the brain of chicken on days 1 and 5 after hatching. PMID- 10674175 TI - Evaluation of cytotoxicity of UHMWPE wear debris. AB - We established a novel method to investigate the phagocytosis of ultra high molecular weight polyethylene using primary macrophage cells by an inverted cell culture method. Abundant wear debris derived from implant materials are generated in aseptic loosening and are deposited in periprosthetic tissues in which they are phagocytized by mono- and multi-nucleated macrophage like cells. Ultra-high molecular-weight-polyethylene wear debris generated from different sources namely, from laboratory test wear machine, in vivo methods and from knee and hip simulator were mainly used in this investigation. The cytotoxicity index of the different UHMWPE particles obtained from various sources were compared with that of the PE beads and the control without particles by Alamar Blue and Neutral Red assays. The results showed that the cytotoxicity index was significantly lower for the wear debris from the in vivo experiments than that for other particles. SEM analysis were also done to understand the morphology of the wear debris and polyethylene beads and to confirm the phagocytosis process. The mean diameter of the wear debris obtained from the in vivo experiments as estimated from the imaging analysis of the SEM photographs was found to be the least. The inverted cell culture method may be regarded as one of the good methods to study the phagocytosis of UHMWPE by macrophage cells. PMID- 10674176 TI - The effect of diameter ratio between vascular substitute and blood vessel on anastomosis. AB - It is necessary to maintain mechanical compatibility between a blood vessel and a vascular substitute to promote encapsulation around the anastomosed part. From this point of view, using linear elastic theory, we had previously performed stress analyses at the part anastomosed by tissue adhesion, in order to propose some methods of preventing stress concentration at this junction. In this study, based on the previous analyses, we have attempted to develop a concept that can be applied under the conditions of operation. That is, the initial diameter of a vascular substitute with high rigidity is chosen larger than that of a blood vessel. This will reduce the stress concentration around the anastomosed part, on average, during expansion of the blood vessel. We analysed the optimum diameter ratio between the vascular substitute and the blood vessel which causes the least stress concentration, on average, during this process, using linear elastic theory. Furthermore, numerical analyses of blood vessel deformation were performed using various nonlinear stress-strain laws. These results were compared to the analytical solution based on linear elastic theory. PMID- 10674177 TI - Dynamics modeling of human temporomandibular joint during whiplash. AB - The objective of this study is to simulate the dynamic response of the temporomandibular joint forces within a rear-end impacted vehicle. Clinicians reports symptoms of temporomandibular joint disorders in many patients who have experienced vehicle rear-end impacts. In rear-end impact, a vehicle occupant's head is thrust rearward with respect to the vehicle in a whiplash action. During this motion, complex dynamic forces act on the jaw bone. To understand the dynamic forces acting on the jaw, we extended an existing human head/neck model by adding a movable jaw, and performed simulation of the jaw motions during rear end impacts at 4.2, 6.4 and 9.6 m/s. Results predicted temporomandibular joint torques, relative angle between the head and jaw, jaw angular acceleration and linear acceleration of the jaw's center of mass. PMID- 10674178 TI - Fatigue strength testing of hip stems with statistical analysis. AB - Component fatigue testing, the final step in the development of total joint replacements, is performed to validate the safety of these components against fatigue failure before clinical use. Fatigue test prediction can aid the design of an efficient fatigue-testing program. The objective of this study was to perform an efficient and accurate statistical analysis of component fatigue test results, for the validation of future fatigue test predictions. Testing was performed with two aims: first, to determine the local component stress-force relationship using strain gauges; and second, to provide a statistical description of the fatigue test results. Forty-nine hip stems, in three sizes, were tested in a series of static and fatigue tests. Through effective planning and analysis, a statistical description of the component fatigue test results was determined including, 3-parameter Weibull distributions of life at two stress levels and log-Normal distributions of fatigue strength at various lives up to 5 million cycles. PMID- 10674179 TI - Apoptosis induced by zinc deficiency in rat osteoblast: possible involvement of protein kinase C. AB - Rat osteoblasts were isolated from the 21-day fetal rat calvarias. The cells were grown in DMEM plus 10% FBS, and were treated for 24 h. with 10 mumol/L TPEN or 10 mumol/L TPEN supplemented with 10 mumol/L Zn2+. Apoptosis of osteoblasts were measured by flow cytometry, electron microscopy and DNA fragmentation analyzed by gel electrophoresis. In addition, IP3 production and PKC activity were measured in order to show whether they are involved in apoptosis in osteoblast induced by zinc deficiency. The results showed that 10 mumol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells treated with 10 mumol/L TPEN supplemented with 10 mumol/L Zn2+ showed no apoptotic changes in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supplemented with Zn2+ simultaneously and the untreated cells. It can be inferred that apoptosis induced by zinc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3. PMID- 10674180 TI - Contribution of an auxin to the uptake of nickel and cadmium in maize seedlings. AB - Maize seedlings were cultured in nickel or cadmium contaminated sand treated with alpha-naphthylacetic acid (NAA). The effects of NAA on nickel and cadmium uptake in roots, shoots, and subcellular fractions (cell wall, nuclei and remained parts of seedling cells) were determined. The data showed growth promotion when NAA was applied at low concentrations and inhibition at high concentrations. Uptake of nickel and cadmium content increased concurrently in roots and shoots. In the subcellular fraction, nickel and cadmium was greatest in the cell wall. The changes in growth had greatest correlation with nickel and cadmium content in the subcellular fraction. PMID- 10674181 TI - Effects of nicotinamide on mouse skin tumor development and its mode of action. AB - Nicotinamide (NA), a naturally occurring vitamin and a protease inhibitor, has been shown to be effective in treating some skin ailments. It inhibits cell proliferation and induces cell differentiation. This report shows the effects of NA on mouse skin tumor development and on the critical events involved in this process. NA reduced tumor growth, inhibited the 12-O-tetradecanoylphorbol-13 acetate (TPA) induced ornithine decarboxylase activity, but induced the transglutaminase activity which was inhibited by TPA under different experimental conditions. The effects of NA on ornithine decarboxylase (ODC) and transglutaminase (TG) indicated that nicotinamide (NA) probably programmed the cells for their death in the natural course of time, i.e. programmed cell death. This observation indicates that NA might be a better agent for the detailed study and for the better use in prevention of cancer alone or in combination with other drugs. PMID- 10674182 TI - Toxicological impact of benzenehexachloride on the behaviour and neuropathology of Heteropneustes fossilis. AB - Toxicological Impact of Benzenehexachloride on the behaviour and Neuropathology of Heteropneustes fossilis. Organochlorine pesticides are widely used in the vast agricultural fields of Assam, India. Run-offs from treated fields contaminates nearby bodies of water with organochlorine compounds, which are neurotoxic to the ichthyofouna. The present work was designed to study the effect of bezenehexachloride on the behaviour and histopathology of Heteropneustes fossilis, as an experimental model. The experimental fish were exposed to different concentrations of the pesticide for 72 hours. After exposure, the fish exhibited various behavioural changes. Histopathological examination of brain tissue revealed cytopathic and gross histopathological alterations, including necrosis and infractional changes. These results are consistent with the finding that organochlorides cause neurotoxic effects. PMID- 10674183 TI - Factors controlling bioindicators for industrial pollution detection. AB - This study describes the use of algae as potential bioindicators of pollution containing industrial metals. Phytoplanktonic algae varied with waste type and with environmental and growth conditions. In water samples containing ceramic waste Euglenophyta species and Cyclotella sp. (Bacillariophyta) were determined as potential indicator species of pollution, while in sample containing metallic waste, Cyclotella sp. was most dominant. Under laboratory growth conditions, phytoplankton collected from a major stream of the Nile River were cultivated by using Algal Growth Bottle Test (EPA, 1972). This revealed that Scenedesmus sp., Actinastrum hantzschii (Chlorophyta), Oscillatoria limnetica (Cyanophyta) and Nitzschia linearis (Bacillariophyta) were also potential indicators of pollution. PMID- 10674184 TI - Histomorphological and histochemical alterations following short-term inhalation exposure to sulfur mustard on visceral organs of mice. AB - Toxic effects of inhaled sulfur mustard (SM) on the histology of visceral organs was investigated by exposing mice to 84.6 mg/m3 for 1 h duration, using controlled single exposure conditions. A progressive fall in body weight from third day onwards was noticed. Light microscopic examination of the pulmonary tissue of these animals at 6 h post exposure revealed that the tracheobronchial epithelium remained intact, but was infiltrated by inflammatory cells. By 24 h post exposure, the mucosecretory cells were destroyed. The inflammatory reaction was maximum at 48 h. By 7th day post exposure there was swelling and vacuolation of lung parenchymal cells and thrombi formation. In addition SM caused congestion and hemorrhage at alveolar level. SM also caused granulovacuolar degeneration with perinuclear clumping of the cytoplasm of hepatocytes and renal parenchymal cells. Renal lesions were characterized by congestion and hemorrhage. Among visceral tissues, maximum atrophy was observed in spleen. Distribution of lesions increased with post exposure period. The maximum lesions were observed at 7th day post-exposure. PMID- 10674185 TI - Influences of chloropazine, nimodipine and their combination on the toxic effects of cadmium in liver and kidney of mice. AB - The influences of the calmodulin antagonist chlorpromazine (CPZ), and calcium channel blocker nimodipine (NIMO) and their combination on cadmium (Cd) poisoning of mice were studied. A series of biochemical parameters including urinary enzyme activities, blood and urine Cd levels, metallothionein (MT) contents in liver and kidney, hepatic ultrastructure and Ca(2+)-Mg2+ ATPase activity in erythrocyte membrane were determined. Animal models for Cd poisoning were established by peritoneal injection of 1/5 LD50 CdCl2. The experimental groups were protected by administration of CPZ, NIMO and CPZ and NIMO in combination 1 h before the injection of CdCl2. Five days later, samples were collected for analysis. The data showed that CPZ could protect kidney tissue against Cd-induced damage, as the urinary gamma-glutamyl-traspeptidase (gamma-GT) and N-acetyl-beta-D glucosaminidase (NAG) activities were reduced significantly. There was neither evidence of the protective effect of NIMO on kidney tissue nor an indication of a synergistic effect of CPZ and NIMO. Both CPZ and NIMO showed a considerable protective effect against the decrease in Ca(2+)-Mg2+ ATPase activity, and a synergistic action was observed. Cd content in blood was reduced significantly by CPZ or the combination of CPZ and NIMO, but elevated by NIMO. Both CPZ and NIMO considerably increased MT contents in livers and kidneys and ameliorated damaged to the hepatic ultrastructures caused by Cd. The results indicated that these inhibitors could protect mice against the toxic effects of Cd in liver and kidney tissues, while CPZ was more efficient than NIMO. The combination of CPZ and NIMO exerted a synergistic action. The protective action of these two drugs might be relevant to the function of MT. PMID- 10674186 TI - A preliminary investigation of the possible hypoglycemic activity of Hibiscus rosa-sinensis. AB - The hypoglycemic activity of an ethanol extract of Hibiscus rosa-sinensis was studied in glucose located rats. After a single dose of the extract, a slight but insignificant hypoglycemic effect was observed at 30 and 90 min. At 120 min it was mild but significant. After repeated administration of the extract (once a day for seven consecutive days) a statistically significant (P < 0.001) reduction in blood glucose levels was observed at 30, 90 and 120 min after glucose loading. The average hypoglycemic activity, after repeated administration of 250 mg kg-1 leaf extract was 81%, under similar conditions average activity of tolbutamide was 96%. At 250 mg.kg-1 the efficacy of the extract was found to be 84% of tolbutamide (100 mg.kg-1). Repeated treatment of animals either with tolbutamide a sulphonylurea or H. rosa-sinensis caused a 2-3-fold improvement in glucose tolerance as compared to those receiving only once. These data suggest that the leaf extract acts like tolbutamide and the mechanism of action may be a stimulation of pancreatic beta cells to produce more insulin or an increase of the glycogen deposition in liver. It appears that the active principle in the tested extract has the sulphonylurea skeleton in which-SO2-NH-CO-group and the substituents (S1 and S2) may be the possible active sites responsible for its hypoglycemic activity. PMID- 10674187 TI - A review: progress of prevention and control on viral hepatitis in China. PMID- 10674188 TI - Anatomic and molecular principles of psychopharmacology. A primer for psychiatrists. AB - Psychopharmacology uses chemicals to modulate human brain function. Three basic principles of neurotransmission may help to understand the current practice of clinical psychopharmacology. First, the anatomic organization of neurotransmitter systems determines their behavioral affiliation. Second, neurotransmitter receptors modulate the electrical properties (via ion channels) or the biochemical properties (via second-messenger systems) of neurons. Third, the intracellular integration of receptor-mediated responses leads to immediate or delayed effects on neuronal function. PMID- 10674189 TI - Pharmacogenetics. Promise and potential in child and adolescent psychiatry. AB - The range of recent promising clinical applications of pharmacogenetics, the nature of genetic research on pharmacokinetic and pharmacodynamic issues, the possible contribution of pharmacogenetics to understanding pathogenesis, and the appropriate genetic methods to be employed are reviewed in this article. Following an overview of the research on the role of dopamine- and serotonin related alleles in influencing risk to neuropsychiatric disorders, separate sections then review the specific pharmacogenetic studies examining the effects of dopamine- and serotonin-related genes on drug response. The potential use of pharmacogenetics in child and adolescent psychopharmacology and the possible directions for future research also are discussed. PMID- 10674190 TI - Cytochrome P450-mediated drug interactions. AB - In this article, the authors have provided child psychiatrists with cytochromal concepts, illustrations of common CYP-based drug interactions, and CYP tables. Clinicians can use these tables to anticipate drug interactions. If two medications are listed on the same CYP, a drug interaction may occur, and depending on whether they are substrates, inducers, or inhibitors, clearance of one or both drugs may be altered. Because new information about CYPs rapidly becomes available, however, CYP tables have a short shelf life. To further predict and reduce the consequences of CYP-based drug interactions, child psychiatrists can limit their own formularies and review PubMed, Ovid, or other literature tracking programs each time they use two or more drugs (including nonpsychiatric ones). The following Internet websites can provide current CYP data: CYP charts http//:@www.dml.georgetown.edu/depts/ph armac ology/clinlist.html http//:@www.accp.com/p450.html CYP drug interaction program http//:@www.mhc.com/Cytochromes/ AIDS drug interactions http//:@www.tthhivclinic.com/interactions.htm http//:@www.fda.gov/oashi/aids/pitabv. htm l http//:@HIV.medscape.com/Medscape/HIV/DrugInteract+ ++ ion s/index.html http//:@www.hopkins-aids.edu/geneva/hilites_f le x_d rug.html. PMID- 10674191 TI - Pharmacotherapy of attention deficit hyperactivity disorder. AB - Despite a large body of literature documenting the effectiveness of medication in the treatment of ADHD, there has been public and professional concern regarding the possible inappropriate diagnosis and prescription of ADHD medications. Recently the Council of Scientific Affairs of the American Medical Association addressed these concerns in a scholarly review. Several factors were identified that contributed to existing controversies: (1) Like most psychiatric disorders, diagnostic criteria for ADHD are based on history and behavioral assessment. There are no pathognomonic laboratory or radiologic tests to confirm the diagnosis. (2) Attention deficit hyperactivity disorder is a chronic disorder and requires extended treatment. (3) Treatment includes potentially abusable medications. After a review of the voluminous literature, this distinguished panel concluded that ADHD is one of the best researched disorders in medicine; in fact, the overall data on its validity are far more compelling than for many other medical conditions. They also concluded that there was little evidence of widespread overdiagnosis or misdiagnosis of ADHD or of widespread overprescription of stimulants by physicians. Consistent with the current emphasis on cognitive dysregulation in ADHD, treatment concerns have expanded from a primarily behavioral focus to include enhancement of executive functions in scholastic as well as other settings. Although stimulants have been the most studied compounds, there is a considerable body of literature indicating an important role for other psychopharmacologic agents. Noradrenergic and dopaminergic modulation appears to be necessary for effective anti-ADHD treatment. In addition, promising evidence of newer cholinergic agents may provide other useful alternatives. As with all psychiatric disorders, comorbid conditions are prominent and may lead to high morbidity and disability if not addressed. As with other areas of medicine, it is sometimes necessary to use multiple agents to treat comorbidity or to achieve an effective response. PMID- 10674192 TI - Pharmacologic treatment of tic disorders. AB - The approach to treating children and adolescents with tic disorders has evolved in recent years such that complete elimination of tics is no longer the primary goal of treatment. Indeed, given the high frequency of psychiatric comorbidity in TS, treatment planning begins with identification of target symptoms. Although traditional neuroleptics still represent standard treatment for tics, many families and clinicians are reluctant to use these agents because of concern about the potential for short- and long-term side effects. Thus, there is great interest in the newer atypical neuroleptics. Interest in the atypical neuroleptics is understandable, but much more study is needed before these agents can become first-line treatments for tics. A small group of non-neuroleptic medications have been used in the treatment of tics. Of these, clonidine, guanfacine, tetrabenazine, pergolide, and botulinum toxin injections have shown some promise for suppressing tics. To date, however, only clonidine has been evaluated in randomized, controlled trials, and the results are not consistent across studies. Although comorbid ADHD is common in children with TS, treatment with stimulant medications was not recommended in children with tics. Recent data suggest that stimulants may be used in some children with TS without adverse effects. Until more is known about which children with ADHD and tic disorders can be safely treated with stimulants, however, the use of stimulants in this population should be undertaken with caution. A handful of nonstimulant medications have been used in the treatment of ADHD with some success, but more study is needed for most of these agents. Evaluation of the stimulants and nonstimulants for the treatment of ADHD in children and adolescents with tic disorders is an area worthy of large controlled trials. PMID- 10674194 TI - Pharmacotherapy of early-onset depression. Update and new directions. AB - Although an increased recognition of depressive disorders in youth represents a positive conceptual change over the past decades, there still is a very limited amount of research on useful treatment interventions. The paucity of data is particularly keen for the use of psychotropic drugs. For example, by applying the criteria suggested by the International Psychopharmacology Algorithm Project, there barely are enough first-grade ("Level A," meaning at least two RCTs) data supporting the short-term efficacy of antidepressants (the SSRIs) in the treatment of juvenile depression. And yet, limited data have not translated into limited use in routine clinical practice. In fact, the use of antidepressant medications has increased exponentially over the last decade, a change that is especially conspicuous for individuals less than 18 years of age. The perceived safety of the SSRIs and other novel antidepressants is partly at the root of their increased popularity. Data regarding their safety are likewise quite limited, however, and essentially are nonexistent for longer-term use. Based on the reviewed data, a medication algorithm for the treatment of early-onset depression can be suggested (Fig. 1). The algorithm underscores the need for adequate evaluation and diagnostic assessment, with particular attention to comorbid conditions (such as a bipolar diathesis) that may dictate alternative treatment strategies. In general, psychotherapy is the initial approach to juvenile MDD, with medication use reserved for more severe cases or those not responding to psychotherapy alone. Given that only two types of psychotherapy and two SSRIs have adequate controlled short-term efficacy data, all but the initial steps must be undertaken guided by clinical judgment and an individualized risk benefit analysis. An algorithm such as this one, based on the very limited efficacy and safety data available, may be viewed as setting priorities for a comprehensive research agenda, more than dictating rigid treatment guidelines. In closing, it can be suggested that future research on the pharmacotherapy of early onset depressive disorder pay particular attention to the following three aspects: 1. Too many drugs, too few data: Rapid advances in drug development have led to a plethora of available antidepressant agents. It is clear that there are many more agents available than can be adequately studied at present. Because many such agents are mechanistically similar, if not identical, it may be wise to focus research efforts on truly novel agents, particularly those (such as the CRH receptor antagonists, or those affecting neurosteroidogenesis) whose action is based on preclinical and clinical pathophysiologic disease paradigms. 2. Longitudinal follow-up and maintenance studies: Essentially all reviewed treatment studies have been short-term trials. There is a marked paucity of longer-term follow-up data, or of naturalistic and "real-world" effectiveness studies. For example, one of the few studies addressing maintenance pharmacotherapy for early-onset depression has demonstrated surprisingly high recurrence rates, even for those subjects actively on maintenance medication. 3. Long-term safety: Clinicians and parents alike often face difficult decisions regarding the long-term exposure of antidepressant drugs on the developing brain. Although no definitive long-term safety data are likely to become available anytime soon, real risks, such as suicide, and potential sequelae of long-term exposure to the underlying illness itself need all to be part of any decision making process. Preclinical studies have shown that brain-derived neurotrophic factor (BDNF) levels can be upregulated by antidepressants, and low BDNF factors have been associated with atrophic brain changes in recurrent forms of adult MDD. Although these observations require specific application to juvenile forms of the disorder, they raise the exciting prospect that the natural course of the illne PMID- 10674193 TI - Pharmacologic treatment of anxiety disorders in children and adolescents. AB - This article reviews the pharmacologic treatment of anxiety disorders in children and adolescents. These disorders are quite common and can be considered a "silent epidemic" because they are more often reported by the children and adolescents than by their parents. Tricyclic antidepressants (TCAs), benzodiazepines, buspirone, and selective serotonin reuptake inhibitors (SSRIs) have been used to treat anxiety disorders in children and adolescents with varying degrees of success. Considering safety and efficacy, the SSRIs appear to be the first-line treatment for anxiety disorders in youth, but more studies are needed to confirm preliminary results. Tricyclic antidepressants and benzodiazepines may be considered when the child has not responded to SSRIs or when adverse effects have exceeded benefits. Although nonpharmacologic approaches for the treatment of anxiety in children and adolescents are beyond the scope of this article, their importance is to be underscored and they should be considered as part of the treatment plan. Over the next decade, research data will be generated regarding the treatment of anxiety disorders in youth. Ongoing research studies include the use of fluoxetine (B. Birmaher, personal communication, 1999) and fluvoxamine (J. Walkup, personal communication, 1999) for the treatment of generalized anxiety disorder, separation anxiety disorder, or social phobia; and buspirone for generalized anxiety disorder in children. Despite these efforts, there is a need for more studies to examine the safety and efficacy of different pharmacologic treatments, as well as longitudinal studies to monitor for long-term tolerability and side effects. Pharmacokinetic studies for children and adolescents will provide information on the metabolism and absorption of these medications and delineate the developmental differences between children and adolescents when compared to adults. Finally, and perhaps most importantly, studies that compare medication, psychosocial treatments, and their combination are needed. PMID- 10674195 TI - Mood stabilizers in the treatment of juvenile bipolar disorder. Advances and controversies. AB - Controlled studies of mood stabilizer (mono and combination) therapy are needed in children and adolescents to develop safe and effective treatment strategies for a disorder that now has a cohort and that carries a high human and economic cost. Through the use of a variety of diagnostic instruments and novel outcome measures, we may continue to refine DSM categories into more sensitive and specific diagnostic constructs. In addition, identification of neurobiologic and genetic markers for early-onset BPD, ADHD, CD, and IED could provide powerful tools in the process of breaking down phenotypes and establishing biologic predictors of targeted pharmacologic interventions in the face of new drug developments. PMID- 10674196 TI - The diagnosis and treatment of children and adolescents with schizophrenia. "My mind is playing tricks on me". AB - This article discusses the clinical phenomenology, natural history, neurobiologic features, diagnostic and medical assessment, and management of schizophrenia, and also details pharmacologic treatments. PMID- 10674197 TI - Research Units on Pediatric Psychopharmacology (RUPP) Autism Network. Background and rationale for an initial controlled study of risperidone. AB - This article has reviewed the background and rationale for the choice of risperidone as the first drug to be studied by the RUPP Autism Network. Risperidone has potent effects on 5-HT and DA neuronal systems, both of which have been implicated in the pathophysiology of autism. Unlike the typical antipsychotics, haloperidol and pimozide, which have been shown to be effective for reducing many of the maladaptive behaviors associated with autism, risperidone's 5-HT2A/DA D2 ratio of receptor blockade appears to produce a lower risk of acute and chronic extrapyramidal side effects, as well as enhanced efficacy for the "negative" symptoms of autism. Indirect clinical and preclinical evidence supports the use of risperidone to treat impaired social behavior, interfering repetitive phenomena, and aggression, targets of pharmacotherapy for many patients with autism. Numerous published open-label trials in children and adolescents with autism and related PDDs and one double-blind, placebo-controlled study in adults suggest that risperidone has promise for the treatment of children and adolescents with autism. Because most of these studies have been short-term, open-label trials in small samples, however, a large-scale controlled study of risperidone in children and adolescents with autism is needed to confirm these results. Finally, because it is likely that children who demonstrate short term benefit from risperidone will remain on the medication indefinitely, the longer-term effectiveness and safety of risperidone in this population also needs to be determined. The design of this study and the assessments used are described separately. PMID- 10674198 TI - Pharmacologic management of psychiatric and behavioral symptoms in mental retardation. AB - Compared with the general population, individuals with mental retardation demonstrate more susceptibility to psychiatric illness and may display disruptive behaviors. These symptoms significantly can affect an already compromised ability to function and the patient may benefit from pharmacologic intervention. Clinical characteristics of individuals with mental retardation warrant special consideration regarding diagnosis and treatment of their psychiatric and behavioral problems. This article describes the nature of symptoms that are typically the target of pharmacologic intervention, outlines special diagnostic considerations, and examines recent findings and experience with psychotropic medication in mental retardation. PMID- 10674199 TI - Practical clinical issues regarding child and adolescent psychopharmacology. AB - The psychotropic treatment of child and adolescent psychiatric disorders is becoming increasingly common. In many cases, its clinical use outstrips its demonstrated scientific validity. Pharmacologic compounds need to be properly utilized, effectively prescribed, and appropriately monitored. This entails a detailed knowledge of various psychotropic agents, their pharmacokinetics and pharmacodynamics, and a high degree of psychotherapeutic sophistication in terms of the patient-family-physician relationship. PMID- 10674200 TI - Lycopene in tomatoes: chemical and physical properties affected by food processing. AB - Lycopene is the pigment principally responsible for the characteristic deep-red color of ripe tomato fruits and tomato products. It has attracted attention due to its biological and physicochemical properties, especially related to its effects as a natural antioxidant. Although it has no provitamin A activity, lycopene does exhibit a physical quenching rate constant with singlet oxygen almost twice as high as that of beta-carotene. This makes its presence in the diet of considerable interest. Increasing clinical evidence supports the role of lycopene as a micronutrient with important health benefits, because it appears to provide protection against a broad range of epithelial cancers. Tomatoes and related tomato products are the major source of lycopene compounds, and are also considered an important source of carotenoids in the human diet. Undesirable degradation of lycopene not only affects the sensory quality of the final products, but also the health benefit of tomato-based foods for the human body. Lycopene in fresh tomato fruits occurs essentially in the all-trans configuration. The main causes of tomato lycopene degradation during processing are isomerization and oxidation. Isomerization converts all-trans isomers to cis isomers due to additional energy input and results in an unstable, energy-rich station. Determination of the degree of lycopene isomerization during processing would provide a measure of the potential health benefits of tomato-based foods. Thermal processing (bleaching, retorting, and freezing processes) generally cause some loss of lycopene in tomato-based foods. Heat induces isomerization of the all-trans to cis forms. The cis-isomers increase with temperature and processing time. In general, dehydrated and powdered tomatoes have poor lycopene stability unless carefully processed and promptly placed in a hermetically sealed and inert atmosphere for storage. A significant increase in the cis-isomers with a simultaneous decrease in the all-trans isomers can be observed in the dehydrated tomato samples using the different dehydration methods. Frozen foods and heat sterilized foods exhibit excellent lycopene stability throughout their normal temperature storage shelf life. Lycopene bioavailability (absorption) can be influenced by many factors. The bioavailability of cis-isomers in food is higher than that of all-trans isomers. Lycopene bioavailability in processed tomato products is higher than in unprocessed fresh tomatoes. The composition and structure of the food also have an impact on the bioavailability of lycopene and may affect the release of lycopene from the tomato tissue matrix. Food processing may improve lycopene bioavailability by breaking down cell walls, which weakens the bonding forces between lycopene and tissue matrix, thus making lycopene more accessible and enhancing the cis-isomerization. More information on lycopene bioavailability, however, is needed. The pharmacokinetic properties of lycopene remain particularly poorly understood. Further research on the bioavalability, pharmacology, biochemistry, and physiology must be done to reveal the mechanism of lycopene in human diet, and the in vivo metabolism of lycopene. Consumer demand for healthy food products provides an opportunity to develop lycopene-rich food as new functional foods, as well as food-grade and pharmaceutical-grade lycopene as new nutraceutical products. An industrial scale, environmentally friendly lycopene extraction and purification procedure with minimal loss of bioactivities is highly desirable for the foods, feed, cosmetic, and pharmaceutical industries. High-quality lycopene products that meet food safety regulations will offer potential benefits to the food industry. PMID- 10674201 TI - Fish protein hydrolysates: production, biochemical, and functional properties. AB - Considerable amounts of fish processing byproducts are discarded each year. By developing enzyme technologies for protein recovery and modification, production of a broad spectrum of food ingredients and industrial products may be possible. Hydrolyzed vegetable and milk proteins are widely used food ingredients. There are few hydrolyzed fish protein foods with the exception of East Asian condiments and sauces. This review describes various manufacturing techniques for fish protein hydrolysates using acid, base, endogenous enzymes, and added bacterial or digestive proteases. The chemical and biochemical characteristics of hydrolyzed fish proteins are discussed. In addition, functional properties of fish protein hydrolysates are described, including solubility, water-holding capacity, emulsification, and foam-forming ability. Possible applications of fish protein hydrolysates in food systems are provided, and comparison with other food protein hydrolysates where pertinent. PMID- 10674202 TI - Toward practical definitions of quality for food science. AB - A new practical approach to developing workable definitions of quality is presented to overcome the numerous semantic and conceptual difficulties that are common with the use of the word quality in food science. This approach links the concept of quality, through a general definition, by adding the missing link of specific definitions related to measurable attributes and properties determined by standard methods to provide values that can be used to evaluate foods or to set specifications. It is compatible with control, assurance, HACCP, regulatory, TQM, and other normal uses of the both the word quality, and the concept quality, in food science and technology. PMID- 10674203 TI - Health care for the homeless: a model for nursing education. PMID- 10674204 TI - The deadly return of tuberculosis. PMID- 10674205 TI - South Africa responds to a TB emergency. PMID- 10674206 TI - A strong nursing role in tuberculosis control. PMID- 10674207 TI - How new mothers acquire knowledge in a hospital setting. PMID- 10674208 TI - Teaching collective health in nursing schools. PMID- 10674209 TI - NMR in biotechnology. PMID- 10674210 TI - Measurement of intracellular (compartmental) pH by 31P NMR in Aspergillus niger. AB - 31P nuclear magnetic resonance (31P NMR) was used to monitor cytoplasmic and vacuolar pH values in the filamentous fungus Aspergillus niger. To obtain a homogeneous cell sample and to be able to perform long term in vivo NMR measurements A. niger mycelium was kept in a setup that allows perfusion of the cell plug within the NMR tube. Mycelial samples, however, became rapidly clogged during perfusion leading to (partial) anaerobiosis of the plug with subsequent acidification of the cytoplasm. As a result, only short-term NMR measurements (5 10 min) were possible using free mycelium. To increase and to prolong perfusion, A. niger was immobilized in Ca(2+)-alginate beads. Deteriorated spectra recorded under hypoxia could be completely restored in the presence of oxygen. With this system perfusion in the presence of citrate could be maintained for at least 18 h at much higher rates (15 ml min-1 compared with 4 ml min-1 for free mycelium). During this period 31P NMR spectra were highly invariable, indicating approximate steady-state intracellular conditions during long term measurements. Perfusion in the presence of glucose resulted in complete depletion of the vacuolar inorganic phosphate pool within 45 min and yielded a higher pH gradient over the tonoplast than when citrate was used (delta pH = 1.6 and 1.4, respectively). PMID- 10674211 TI - Analysis of sugar metabolism in an EPS producing Lactococcus lactis by 31P NMR. AB - Sugar metabolism and exopolysaccharide (EPS) production was analysed in Lactococcus lactis by in vivo 31P NMR. Transient production of several sugar phosphates, transient depletion of intracellular phosphate, transient production of ATP and UTP, transient acidification of the medium and alkalinisation of the cytoplasm could be observed in a period of 20 min upon energization by the addition of glucose. EPS and non-EPS producing variants showed similar NMR spectra, the exception being two pH-dependent resonances observed in the former. They were already observed before addition of glucose and their response to glucose incubation reflected exposure to the medium. They are presumably phosphorylated poly- or oligosaccharides being loosely adhered to cell walls. By freezing and perchloric acid extraction of the cell material, different types of phosphorylated compounds could be recognised in the NMR spectra such as fructose 1-6-diphosphate, nucleotides (like ADP, ATP, UTP and TDP) and several nucleotide sugars. The ongoing work is focused on identifying the unknown peaks and quantifying the differences between wild-type cells and the EPS producing variant. PMID- 10674212 TI - Determination of full 13C isotopomer distributions for metabolic flux analysis using heteronuclear spin echo difference NMR spectroscopy. AB - 13C-isotopomer labeling experiments play an increasingly important role in the analysis of intracellular metabolic fluxes for genetic engineering purposes. 13C NMR spectroscopy is a key technique in the experimental determination of isotopomer distributions. However, only subsets of isotopomers can be quantitated using this technique due to redundancies in the scalar coupling patterns and due to invisibility of the 12C isotope in NMR. Therefore, we developed and describe in this paper a 1H NMR spectroscopy method that allows to determine the complete isotopomer distribution in metabolites having a backbone consisting of up to at least four carbons. The proposed pulse sequences employ up to three alternately applied frequency-selective inversion pulses in the 13C channel. In a first application study, the complete isotopomer distribution of aspartate isolated from [1-13C]ethanol-grown Ashbya gossypii was determined. A tentative model of the central metabolism of this organism was constructed and used for metabolic flux analysis. The aspartate isotopomer NMR data played a key role in the successful determination of the flux through the peroxisomal glyoxylate pathway. The new NMR method can be highly instrumental in generating the data upon which isotopomer labeling experiments for flux analysis, that are becoming increasingly important, are based. PMID- 10674213 TI - 13C and 1H NMR study of cellulose metabolism by Fibrobacter succinogenes S85. AB - Fibrobacter succinogenes S85, a cellulolytic rumen bacterium, is very efficient in degrading lignocellulosic substrates and could be used to develop a biotechnological process for the treatment of wastes. In this work, the metabolism of cellulose by F. succinogenes S85 was investigated using in vivo 13C NMR and 13C-filtered spin-echo difference 1H NMR spectroscopy. The degradation of unlabelled cellulose synthesised by Acetobacter xylinum was studied indirectly, in the presence of [1-13C]glucose, by estimating the isotopic dilution of the final bacterial fermentation products (glycogen, succinate, acetate). During the pre-incubation period of F. succinogenes cells with cellulose fibres, some cells ('non-adherent') did not attach to the solid material. Results for 'adherent' cells showed that about one fourth of the glucose units entering F. succinogenes metabolism originated from cellulose degradation. A huge reversal of succinate metabolism pathway and production of large amounts of unlabelled acetate which was observed during incubation with glucose only, was found to be much decreased in the presence of solid substrate. The synthesis of glucose 6-phophate was slightly increased in the presence of cellulose. Results clearly showed that 'non adherent' cells were able to metabolise glucose very efficiently; consequently the metabolic state of these cells was not responsible for their 'non-adherence' to cellulose fibre. PMID- 10674214 TI - Relevance and isotopic assessment of hexose-6-phosphate recycling in micro organisms. AB - Some pathways of hexose-6-phosphate recycling--those involving a breakdown of the hexose skeleton--through carbohydrate metabolism of micro-organisms were analyzed for both metabolic and isotopic effects. Two modes of recycling were proposed based on the degree of alteration of the hexose molecule through the catabolic part of the cycle. Simulated operation of most of these pathways resulted in increased synthesis of hexose-6-phosphate and NADPH, and reduced the NADH and moreover the ATP synthesis within the carbohydrate metabolism. A basic model for the quantitative assessment by means of isotopic studies of the processes of hexose-6-phosphate recycling is presented. The model was initially designed for the study of micro-organisms producing polysaccharides, but it can be extended to other situations. PMID- 10674215 TI - Introduction of polyphosphate as a novel phosphate pool in the chloroplast of transgenic potato plants modifies carbohydrate partitioning. AB - Potato plants (Solanum tuberosum L., cv. Desiree) were transformed with the polyphosphate kinase gene from Escherichia coli fused to the leader sequence of the ferredoxin oxidoreductase gene (FNR) from Spinacea oleracea under the control of the leaf specific St-LS1 promoter to introduce a novel phosphate pool in the chloroplasts of green tissues. Transgenic plants (cpPPK) in tissue culture developed necrotic lesions in older leaves and showed earlier leaf senescence while greenhouse plants showed no noticeable phenotype. Leaves of cpPPK plants contained less starch but higher concentrations of soluble sugars. The presence of polyphosphate in cpPPK leaves was demonstrated by toluidine blue staining and unambiguously verified and quantified by in vitro 31P-NMR of extracts. Polyphosphate accumulated during leaf development from 0.06 in juvenile leaves to 0.83 mg P g-1 DW in old leaves and had an average chain length of 18 residues in mature leaves. In situ 31P-NMR on small leaf pieces perfused with well-oxygenated medium showed only 0.036 mg P g-1 DW polyphosphate that was, however, greatly increased upon treatment with 50 mM ammonium sulfate at pH 7.3. This phenomenon along with a yield of 0.47 mg P g-1 DW polyphosphate from an extract of the same leaf material suggests that 93% of the polyphosphate pool is immobile. This conclusion is substantiated by the observation that no differences in polyphosphate pool sizes could be discerned between darkened and illuminated leaves, leaves treated with methylviologen or anaerobis and control leaves, treatments causing a change in the pool of ATP available for polyPi synthesis. Results are discussed in the context of the chelating properties of polyphosphates for cations and its consequences for the partitioning of photoassimilate between starch and soluble sugars. PMID- 10674216 TI - Strategies for metabolic flux analysis in plants using isotope labelling. AB - Flux measurements through metabolic pathways generate insights into the integration of metabolism, and there is increasing interest in using such measurements to quantify the metabolic effects of mutation and genetic manipulation. Isotope labelling provides a powerful approach for measuring metabolic fluxes, and it gives rise to several distinct methods based on either dynamic or steady-state experiments. We discuss the application of these methods to photosynthetic and non-photosynthetic plant tissues, and we illustrate the different approaches with an analysis of the pathways interconverting hexose phosphates and triose phosphates. The complicating effects of the pentose phosphate pathway and the problems arising from the extensive compartmentation of plant cell metabolism are considered. The non-trivial nature of the analysis is emphasised by reference to invalid deductions in earlier work. It is concluded that steady-state isotopic labelling experiments can provide important information on the fluxes through primary metabolism in plants, and that the combination of stable isotope labelling with detection by nuclear magnetic resonance is particularly informative. PMID- 10674217 TI - Chemical fingerprinting for the evaluation of unintended secondary metabolic changes in transgenic food crops. AB - A common element in designed guidelines for assessment of the food safety of transgenic crops is centred on a comparative analytical analysis with conventionally bred crop plants, assuming that these products have a long history of safe use (i.e. OECD-principle of substantial equivalence). In this study we examine the utility of an off-line combination of 400 MHz proton (1H)-NMR spectroscopy and liquid chromatography (LC) for the multi-component comparison of low-molecular weight compounds (i.e. chemical fingerprinting) in complex plant matrices. The developed NMR-methodology can contribute to the demonstration of substantial equivalence by its ability to compare possible compositional alterations in a novel food crop with respect to related non-transgenic reference lines. In this respect a hierarchical approach is proposed by comparing the chemical fingerprints of the transgenic crop plant to those of: (1) isogenic parental or closely related lines bred at identical and multiple sites; (2) extended ranges of commercial varieties of that plant; and (3) downstream processing effects. This is of importance to assess the likelihood that some of the statistical differences in a transgenic crop plant may be false positives due to chance alone or arose from natural genetic and/or physiologic variations. PMID- 10674218 TI - Structure elucidation of glycoprotein glycans and of polysaccharides by NMR spectroscopy. AB - The applicability of 1H-NMR spectroscopy for the determination of the primary and tertiary structure of carbohydrate-containing molecules is demonstrated. For classes of known compounds the characterization can be based on chemical shifts observed in 1D NMR spectra with or without the aid of a computer database. For more complex structure determinations 2D NMR techniques are required. Here the application of 2D NMR is demonstrated for the primary structure determination of two bacterial exopolysaccharides, for the spatial structure determination of a disaccharide and a glycoprotein hormone. PMID- 10674219 TI - Structure/function studies of anticoagulant sulphated polysaccharides using NMR. AB - Sulphated polysaccharides have many biological functions, which depend on binding of highly specific carbohydrate structures to proteins. NMR spectroscopy is a technique capable of detailed structural elucidation of these polysaccharides, and can be used in applications ranging from routine analysis to research into covalent and conformational aspects of polysaccharide structure. This technique can be used to characterise sequence variations in heparin samples. The NMR determined solution conformation of heparin has been used to predict binding sites on the surface of heparin-binding proteins. Sulphation patterns for dermatan sulphates of marine invertebrates have been determined. Their anticoagulant effects depend on an exact pattern of sulphate substitution. A small alteration in dermatan sulphate structure, from 4-O-sulphated to 6-O sulphated galactosamine, leads to almost complete loss of anticoagulant activity in spite of an overall high level of sulphation. A fucosylated chondroitin sulphate isolated from sea cucumber has anticoagulant and antithrombotic activity depending on its sulphated fucose branches. The anticoagulant activity of algal fucans has been compared with that of regular, linear sulphated fucans from marine echinoderms; again high activity appears to correlate with the presence of sulphated fucose branches. PMID- 10674220 TI - Diffusion in Pseudomonas aeruginosa biofilms: a pulsed field gradient NMR study. AB - A Pseudomonas aeruginosa biofilm is studied with pulsed field gradient echo nuclear magnetic resonance. Although not all spectral components are assigned yet, the experimental results show that a biofilm consists of components with very different diffusion coefficients. The various biofilm components that give motionally narrowed 1H NMR signals, can be grouped into five classes with diffusion coefficients, ranging from 2 x 10(-9) to 1 x 10(-13) m2 s-1. Investigation of the diffusion behavior of water in the biofilm shows three fractions with different diffusion coefficients. Besides the highly mobile bulk water at least two other fractions with much lower diffusion coefficients are detected. It is shown that one of the fractions with the low diffusion coefficient probably arises from intracellular water. Also for another component of the biofilm, glycerol, three fractions with diffusion coefficients that differ more than a factor ten are detected. Also a group of signals exists which result from practically immobile components. PMID- 10674221 TI - [Partial non-convulsive status epilepsy in multiple sclerosis]. AB - This report describes the observations of two patients with a several years' history of multiple sclerosis who presented sudden neurologic impairment. The symptomatology was suggestive of a non-convulsive partial status epilepsy. The clinical presentation was a paroxysmal dysphasic phenomenon in the first case without any consciousness impairment, associated with slight right hemiparesis. Electroencephalographic investigations revealed asymmetrical patterns, left-sided slow waves and periodic lateralized epileptiform discharges (PLEDs). Antiepileptic treatments were partially effective and intravenous steroids were needed for complete recovery. For the second patient, clinical presentation was acute psychiatric symptoms with disorientation, alternating manic symptomatology and mutism. Electroencephalography showed left fronto-central rhythmic continuous slow wave and spike wave activity. Intravenous antiepileptic treatment quickly improved the symptomatology. These observations draw attention to the fact that an epileptic cause should not be ruled out when a patient with multiple sclerosis presents sudden neurologic or psychiatric impairment. An early diagnosis allows immediate antiepileptic treatment. Intravenous steroids can be added to stop seizures. PMID- 10674222 TI - [Determination of language dominance using EEG spectral analysis versus the Wada test in temporal epilepsy (right-handed subjects)]. AB - Spectral analysis of the EEG alpha rhythm was studied in nine temporal epileptic right-handed patients in order to predict localization of the speech area. We studied the variation of the spectral power of the alpha rhythm during an activation paradigm previously validated in normal right-handed subjects. Significant alpha power decreases in the left hemisphere during writing with the right hand (as compared to resting) and/or significant alpha power decreases in the right hemisphere during left-hand recognition and classification of cardboard objects (as compared to resting) were considered as consistent with left hemisphere dominance for language. The results of EEG spectral analysis were compared with those of the Wada test. The left hemisphere was dominant for language according to the Wada test in eight subjects and the right hemisphere in one subject. Six patients had a significant alpha power reduction in the hemisphere concerned during lateralized cognitive tasks, consistent with language localization in the left hemisphere according to the Wada test. The three remaining patients had no significant EEG spectral power variations. A significant decrease of alpha power in the active hemisphere during cerebral activation seems statistically related to left-hemispheric dominance for language in right-handed subjects (hemispheric specialization). However, the localization of the speech area using this electrophysiological method does not appear clinically relevant for a case-by-case decision in individual patients. PMID- 10674223 TI - Acetylcholine receptor activation enhances NMDA-mediated responses in the rat neostriatum. AB - The influence of acetylcholine (ACh) upon N-methyl-D-aspartate (NMDA) receptor activation of neostriatal neurons is unknown. In the present study, we used both in vitro intracellular and in vivo electroencephalographic recordings in rats to examine this question. In vitro, iontophoretic application of carbachol, a cholinergic receptor agonist, significantly increased the NMDA-mediated response of neostriatal projection neurons. Carbachol alone had mild excitatory effects. In vivo, intrastriatal NMDA produced focal epileptiform activity restricted to the neostriatum. NMDA applied in conjunction with carbachol produced significantly greater epileptiform activity which propagated to the neocortex. These results suggest that ACh and NMDA receptor co-activation leads to potentiation of the neuronal responses both at the site of the interaction and at the endpoint of the cortico-striato-cortical circuit. PMID- 10674224 TI - [Pseudo-paralysis of the brachial biceps in obstetrical brachial plexus lesions (OBPL):concerning the "overly optimistic" EMG in OBPL]. AB - In birth palsy of the brachial plexus, the mixed interference pattern recorded for the brachial biceps on the electromyogram often conflicts with the muscle's inability to flex the elbow. We report our observations of a six-month-old infant who presented paralysis of the upper and medial elements of the brachial plexus, in whom we demonstrated early biceps-triceps co-contractions, which may explain this discrepancy and 'pseudo-paralysis' of the biceps. We analyse and discuss the practical consequences of these findings, and notably the possible therapeutic use of triceps-to-biceps surgical transposition. PMID- 10674225 TI - [Olfactory lateralization in humans: a review of the literature]. AB - In the field of human perception, the chemical senses (taste and smell) have received little attention from neuroscience research when compared with auditory, visual and tactile senses. In the case of olfaction, it would appear that the publications over the last few years have been trying to overcome this lack of research. Many investigations have been carried out on lateralization, mainly in relation to specific pathologies (i.e., epilepsy, split-brain, lobotomy, etc.), while there have been few studies of healthy subjects. The results are often contradictory due particularly to special features of the olfactory system. However, consensus is emerging concerning, first, the fact that if both hemispheres are involved in the olfactory process, it is probable that one is more dominant than the other (many studies have revealed a greater impact on the right hemisphere in the treatment of olfactory information, but the dominance has not been clearly established). Second, the simple detection process would appear not to be lateralized whereas the higher-order olfactory tasks which involve memory processes and lexical aspects could be. The exact conditions governing lateralization still require more clarification by systematically taking into consideration the characteristics of the individual subjects, as well as those of the odorant stimuli and the test conditions. Finally, currently available techniques used in neurosciences and particularly cerebral imagery will contribute to a better understanding of the complexity of cerebral asymmetry in olfaction. PMID- 10674226 TI - A multi-center trial of the effects of oral nutritional supplementation in critically ill older inpatients. GAGE Group. Groupe Aquitain Geriatrique d'Evaluation. AB - The purpose of this study was to assess the effect of nutritional supplementation on dietary intake and on pressure ulcer development in critically ill older patients. The multi-center trial involved 19 wards stratified according to specialty and recruitment for critically ill older patients; 9 wards were randomly selected for nutritional intervention (nutritional intervention group), consisting of the daily distribution of two oral supplements, with each supplement containg 200 kcal, for 15 d. Pressure ulcer incidence was prospectively recorded for grades I (erythema), II (superficial broken skin), and III (subcutaneous lesion) for 15 d. Nutritional intake was monitored by using estimates in units of quarters validated by comparison with weight measurement. There were 672 subjects older than 65 y, and 295 were in the nutritional intervention group versus 377 in the control group. The patients were similar for age, sex ratio, and C-reactive protein. In comparison with the control group, the nutritional intervention group included more patients with stroke, heart failure, and dyspnea and fewer with antecedent falls, delirium, lower limb fractures, and digestive disease. The nutritional intervention group had a lower risk of pressure ulcers according to the Norton score but was less dependent (Kuntzman score) and had a lower serum albumin level. During the trial, energy and protein intakes were higher in the nutritional intervention group (day 2: 1081 +/- 595 kcal versus 957 +/- 530 kcal, P = 0.006; 45.9 +/- 27.8 g protein versus 38.3 +/- 23.8 g protein in the control group, P < 0.001). At 15 d, the cumulative incidence of pressure ulcers was 40.6% in the nutritional intervention group versus 47.2% in the control group. The proportion of grade I cases relative to the total number of cases was 90%. Multivariate analysis, taking into account all diagnoses, potential risk factors, and the intra-ward correlation, indicated that the independent risk factors of developing a pressure ulcer during this period were: serum albumin level at baseline, for 1 g/L decrease: 1.05 (95% confidence interval: 1.02 to 1.07, P < 0.001); Kuntzmann score at baseline, for 1-point increase: 1.22 (0.32 to 4.58, P = 0.003); lower limb fracture: 2.68 (1.75 to 4.11, P < 0.001); Norton score < 10 versus > 14: 1.28 (1.01 to 1.62, P = 0.04); and belonging to the control group: 1.57 (1.03 to 2.38, P = 0.04). In conclusion, it was possible to increase the dietary intake of critically ill elderly subjects by systematic use of oral supplements. This intervention was associated with a decreased risk of pressure ulcer incidence. PMID- 10674227 TI - Influence of the 13C-enrichment of the habitual diet on a 13CO2 breath test used as an index of liver glycogen oxidation: a validation study in western Europe and Africa. AB - A diet containing naturally 13C-enriched carbohydrate combined with a 13CO2 breath-test analysis can be used to monitor liver glycogen oxidation in persons used to a diet low in 13C, e.g., the Western European diet. In this study, we evaluated this test principle further by changing the way we label the glycogen pool. The 13C enrichment of exhaled CO2 was studied in two groups, one in Europe and one in Africa. The European group (n = 12) was accustomed to a diet low in 13C, and they went on a 13C-enriched study diet to identify liver glycogen. The African group (n = 6) was accustomed to a diet naturally high in 13C, and they went on a diet low in 13C. The basal 13C abundance in exhaled CO2 was higher in the African group (1.0879 At%; atmospheric 1.1 atom percent) than in the European group (1.0821 At%). During the study period, the parameters for liver glycogen oxidation--the 13CO2 enrichment plateau, the plateau duration, and the return to baseline time--did not differ between groups. The abundance of 13CO2 in exhaled CO2 over time in the two groups was similar but inverse. This study confirms the use of a 13CO2 breath test to monitor liver glycogen oxidation and demonstrates how to use such a test in persons accustomed to a diet high in 13C. PMID- 10674228 TI - Effects of eicosapentaenoic acid intake on plasma fibrinolytic and coagulation activity by using physical load in the young. AB - To assess the effect of eicosapentaenoic acid (EPA) intake on fibrinolysis and coagulation, 30 male subjects, approximately 19-23 y old, were examined for plasma fibrinolytic and coagulation activity by using a bicycle ergometer load (90 W, 20 min) before and after EPA intake of 1.125 g/d for 2 wk. Because of the EPA intake, the fibrinolytic activity was promoted, the plasmin-alpha 2 plasmininhibitor complex (PIC) level was decreased by 16.7%, and the thrombin antithrombin III complex (TAT) level was increased by 75.4%; conversely, the D dimer of the fibrin degradation peptide (D-dimer) level did not change from that before EPA intake. By the physical load, 1 h after ingesting the load, the PIC level was significantly decreased by 26.7%, the TAT level was significantly increased by 51.1%, and the D-dimer level was significantly decreased by 24% in comparison with levels before EPA intake. Thus, as determined by the load, a small amount of daily EPA intake clearly decreased fibrinolytic activity and increased coagulation activity. One hour after a physical load, the rate of change of the gamma-glutamyl transpeptidase (gamma-GTP) level correlated significantly and negatively to the rate of change in the PIC and TAT levels. Thus, EPA intake may affect liver and kidney function. EPA intake decreased systolic blood pressure by 5 mmHg and diastolic blood pressure by 10 mmHg. PMID- 10674229 TI - Differential effects of amino acid and ketoacid on protein metabolism in humans. AB - We examined the effects of insulin, amino acid (AA), and branched-chain ketoacid (KA) availability on leucine kinetics in eight healthy volunteers (age = 22 +/- 2 y, body mass index = 24 +/- 1 kg) by using the euglycemic insulin clamp and [1 14C] leucine turnover techniques. Four experimental conditions were studied: study I, hyperinsulinemia; study II, hyperinsulinemia with maintenance of basal plasma AA and branched-chain KA concentrations; study III, hyperinsulinemia with hyperaminoacidemia and basal plasma branched-chain KA concentrations; and study IV, hyperinsulinemia plus basal plasma AA concentrations and elevated branched chain KA levels. Basal endogenous leucine flux (ELF) averaged 1.20 +/- 0.05 (mumol.kg-1.min-1, mean +/- SE); basal leucine oxidation (LOX) was 0.25 +/- 0.01; and basal non-oxidative leucine disposal (NOLD) was 0.95 +/- 0.04. ELF significantly decreased in study I (0.77 +/- 0.06 mumol.kg-1.min-1, P < 0.01 versus basal). When plasma AA and branched-chain KA were either maintained at their basal levels (study II) or increased above baseline values (studies III and IV), ELF declined further (0.64 +/- 0.05, 0.66 +/- 0.02, and 0.66 +/- 0.03 mumol.kg-1.min-1, respectively; all Ps < 0.01 versus basal and P < 0.01 versus study I). LOX declined in study I (0.12 +/- 0.02 mumol.kg-1.min-1, P < 0.01 versus basal) but increased significantly in studies II, III, and IV (0.31 +/- 0.04, 0.37 +/- 0.03, and 0.40 +/- 0.03 mumol.kg-1.min-1, respectively, all Ps < 0.01 versus basal, P < 0.05 study IV versus study II, and P < 0.05 study III versus study II). NOLD declined in study I (0.65 +/- 0.05 mumol/kg.min, P < 0.01 versus basal), whereas neither the maintenance of basal plasma AA/branched-chain KA levels (study II; 0.89 +/- 0.2 mumol.kg-1.min-1) nor the elevation of plasma branched-chain KA concentration (study IV; 0.96 +/- 0.1 mumol.kg-1.min-1) increased NOLD above baseline level. A stimulation of NOLD was observed only when plasma AA levels were increased (study III; 1.23 +/- 0.03 mumol/kg.min, P < 0.01 versus basal). In conclusion, the present data do not support the concept of a direct anabolic action of ketoanalogs but do provide additional evidence for the pivotal role of AA availability in the stimulation of whole-body protein synthesis. PMID- 10674230 TI - Selenium kinetics and changes in glutathione peroxidase activities in patients receiving long-term parenteral nutrition and effects of supplementation with selenite. AB - Selenium (Se) is an essential trace element in humans. Patients receiving long term parenteral nutrition (PN) are at risk for Se deficiency. We investigated changes in Se levels and glutathione peroxidase (GSH-Px) activity in serum and tissue (red blood cells, RBC) in addition to urinary excretion of Se in patients receiving long-term PN with and without Se supplementation. In patients without Se supplementation, both Se levels and GSH-Px activity in serum decreased with duration of PN. The serum Se levels were below the lower limits of the control values in 19 of 33 patients (58%) who received PN for less than 1 mo. Conversely, RBC GSH-Px activity remained at a sufficient level in 9 of 12 patients (75%) who received PN for 3-6 mo. The RBC Se levels in all of these patients were lower than the control levels. Urinary Se concentrations were significantly correlated with serum Se concentrations by linear regression analysis (r = 0.707, P < 0.05). In patients with Se supplementation, urinary Se concentrations increased exponentially with increases in serum Se levels. These findings indicate that a time lag precedes the decrease in levels of serum Se, RBC Se, serum GSH-Px, and RBC GSH-Px in patients without Se supplementation and the increase in excretion of urinary Se in patients with Se supplementation. The monitoring of not only serum Se levels but also RBC GSH-Px activity and urinary Se levels is required for optimal Se supplementation during long-term PN. PMID- 10674231 TI - Work-shift period and weight change. AB - The present study was done to determine whether weight gain was more prevalent in workers on late shifts than in those on day shifts. A questionnaire about changes in weight, food intake, exercise, and sleep since starting the job on the current shift was given to day-shift and late-shift (evening and night) hospital workers. Data were analyzed for 85 subjects, 36 of whom worked during the day shift and 49 the late shift. The late-shift group reported a mean weight gain of 4.3 kg, which was greater than the mean weight gain of 0.9 kg for the day-shift group (P = 0.02). There were, however, no significant differences in current body mass index (26.7 +/- 5.4 SD) between groups. There was a trend for late-shift workers to report eating more since beginning the later shift (P = 0.06). When combined with those reporting exercising less (P = NS), this trend became significant (P = 0.04). Late-shift workers reported eating fewer meals (1.9 +/- 0.9 SD) than the day-shift workers (2.5 +/- 0.9; P = 0.002). In addition, late-shift workers reported eating the last daily meal later (mean = 22:27, or 10:27 PM) than day shift workers (17:52 or 5:52 PM; P < 0.00005). Late-shift workers also reported more naps (P = 0.01) and longer naps (P = 0.05) during the work week than did day shift workers. The reported changes in eating, exercise, and sleep may contribute to the increased weight gain of late-shift workers. PMID- 10674232 TI - Effect of portal hypertension caused by chronic high venous pressure on small intestinal sugar absorption. AB - The effect of portal hypertension and chronic high venous pressure on the absorption of the small intestine was examined by constricting the suprahepatic and subdiaphragmatic inferior vena cava in rats. A group of rats with the constricted suprahepatic and subdiaphragmatic inferior vena cava comprised group 1 (n = 9) and another group of rats with only laparotomy comprised group 2 (n = 9). Two months after the operation, sugar absorption and other parameters were measured. The blood pressures of the infrahepatic inferior vena cava and the portal vein 8 wk after the operation in group 1 were significantly higher than those in group 2. The results of D-xylose absorption tests showed that the amount of excreted D-xylose in urine in group 1 was significantly lower than that of group 2, but the D-xylose everted sac test showed no significant differences between the two groups. The glucose everted sac test showed that the amount of glucose absorption in group 1 was significantly lower than that in group 2. These findings suggest that chronic high venous pressure caused by constriction of suprahepatic inferior vena cava may lead to sugar malabsorption. We conclude that portal hypertension with high venous pressure below the diaphragm can lead to sugar malabsorption in the intestine. PMID- 10674233 TI - Nitric oxide synthase inhibitor attenuates inflammatory lesions in the skin of zinc-deficient rats. AB - Skin lesions are common manifestations of zinc deficiency in humans and animals, but the pathogenic mechanisms have not been fully clarified. In the present study, a nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (L NAME), was given to zinc-deficient (ZD) rats to see whether it prevents or delays the occurrence of skin lesions. Weanling male rats were given free access to a ZD diet (2 mg zinc/kg) for 4 wk to induce zinc deficiency. Control rats, including pair-fed (PF) and ad libitum (AL) groups, were given a diet supplemented with zinc (50.8 mg zinc/kg. L-NAME (0.3 g/L in drinking water) was given to some ZD rats for 3 wk, starting at the second week of their ZD dieting. Dermatitis of the extremities, balanitis, stomatitis, and alopecia appeared in ZD but not in AL and PF rats. Administration of L-NAME significantly reduced the frequency of cutaneous and mucocutaneous inflammatory lesions but did not prevent alopecia in the ZD rats. Reverse transcription polymerase chain reaction showed that inducible nitric oxide synthase mRNA was expressed in the paw skin of ZD but not of AL and PF rats. Evaluation of skin microvascular permeability by the Evans blue leakage technique indicated that L-NAME administration significantly attenuated extravasation of Evans blue in the paw skin of ZD rats. Furthermore, stains positive for terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling were condensed and diffusely distributed over the epidermis, dermis, and subcutaneous tissue of paws in ZD rats. ZD rats had intense cell infiltration and parakeratosis in the paw skin. L-NAME administration effectively prevented these morphologic changes. These results demonstrate that nitric oxide synthase inhibitor ameliorates inflammatory lesions of the skin in ZD rats. PMID- 10674234 TI - Postprandial resynthesis of myofibrillar proteins is translationally rather than transcriptionally regulated in human skeletal muscle. AB - Feeding stimulates protein synthesis in skeletal muscles, although the regulatory mechanisms are incompletely understood. The aim of this study was to determine whether this could be detected at the gene transcription level for postprandial stimulation of the synthesis of muscle proteins. Healthy male volunteers were investigated after an overnight fast. Open muscle biopsies were performed in the starved state and 3 h after meal intake, consisting of 0.15 gN/kg, 12 kcal/kg. Blood samples were drawn every 15 to 30 min for 5 h. Myosin mRNA and insulin growth factor-I (IGF-I) mRNA were measured by solution hybridization assay in homogenized muscle specimens. After food intake, plasma glucose concentrations increased from 5.0 +/- 0.1 to 7.3 +/- 0.3 (P < or = 0.001), and insulin concentration rose from 3.8 +/- 0.5 mU/L before to 75.3 +/- 11.4 15 min after the meal (P < or = 0.001). Plasma concentration of free fatty acids declined after food intake (P < or = 0.001). Plasma concentrations of amino acids increased from basal values (2864 +/- 128 microM) to 4419 +/- 262 microM (P < or = 0.05) 90 min after meal ingestion. Myosin mRNA concentration in the biopsied muscle tissue was higher during starvation and was reduced by 20% after food intake: 10.8 +/- 1.3 amol mRNA/microgram DNA in the starved state and 8.5 +/- 1.3 amol mRNA/microgram DNA after food intake (P < or = 0.05). Feeding did not alter IGF-I mRNA concentrations in muscle: 0.51 +/- 0.05 and 0.55 +/- 0.06 amol/microgram DNA in the starved and fed state, respectively (P < or = 0.48). Improved protein balance by stimulation of protein synthesis has been related to increased plasma amino acids. Interestingly, in the short term, this was not related to increases in gene transcription of either myofibrillar proteins (myosin) or muscle IGF-I. Thus, postprandial stimulation of protein synthesis appears not to be regulated by increased gene transcription but by increased translation using the increased concentrations of amino acids. In contrast, as far as the 2X myosin mRNA level is concerned, this is enhanced during starvation, which facilitates rapid recovery once the availability of substrate is resumed. PMID- 10674235 TI - Does jejunal feeding with a polymeric immune-enhancing formula increase pancreatic exocrine output as compared with TPN? A case report. AB - This case report compares the pancreatic output with different feeding regimes in a patient who underwent a partial pancreatectomy for carcinoma of the ampulla of Vater. A postoperative secretin stimulation test demonstrated significant pancreatic reserve. There was no difference in pancreatic exocrine secretion when the patient was fed jejunally with a polymeric immune-enhancing formula or supported with two different formulations of total parenteral nutrition. This result suggests that jejunal infusion of a polymeric immune-enhancing formula may be safe to administer in patients with acute pancreatitis. PMID- 10674236 TI - Assessment of protein energy malnutrition in older persons, part I: History, examination, body composition, and screening tools. AB - Protein-energy malnutrition is a prevalent problem in older persons. Its relation to increased morbidity and mortality has been well documented. Early recognition of malnutrition allows for a timely intervention. A large proportion of chronic diseases affecting older persons can be either prevented or significantly improved by improving nutrition, which underscores the importance of developing a screening system that can trigger a more comprehensive evaluation when indicated. Screening for malnutrition in older persons can be difficult because of the normal age-related changes in many of the commonly used parameters. A comprehensive nutritional evaluation includes a complete history and physical examination in addition to a more specific nutrition-oriented assessment. Specific nutritional assessment includes estimating food intake, anthropometric measurements, and evaluation of several biochemical parameters commonly affected by changes in nutritional status. In this article, we review the commonly used tools for nutritional assessment in older persons. The goal is to promote disease free, active, and successful aging. PMID- 10674237 TI - An osmotic stimulus-mediating glucagon-like peptide-1 (7-36 amide) (GLP-1) secretion in acarbose-induced sucrose malabsorption? PMID- 10674239 TI - Of Vikings, Apollo, serendipity, and research letters. PMID- 10674238 TI - Dopamine in the VMN of the hypothalamus is important for diurnal distribution of eating in obese male Zucker rats. PMID- 10674240 TI - Effects of glutamine on immune cells. PMID- 10674241 TI - Effect of dietary glutamate on chemotherapy-induced immunosuppression. PMID- 10674242 TI - Glutamine-enriched enteral nutrition in multiple trauma patients. PMID- 10674243 TI - More good news about glutamine. PMID- 10674244 TI - Arginine and immunonutrition: a reevaluation. PMID- 10674245 TI - Perioperative nutritional support in liver surgery. PMID- 10674246 TI - Nutrition and Crohn's disease: a case study in ambiguity. PMID- 10674247 TI - Nutrition in pediatric inflammatory bowel disease. PMID- 10674248 TI - Regression to the mean. PMID- 10674249 TI - Repair of double-lumen tube with simple and inexpensive materials. PMID- 10674250 TI - The first PEG. PMID- 10674251 TI - Red blood cell membrane oxidative damage and renal impairment in uremic patients under conservative treatment. PMID- 10674252 TI - Pregnancy in the postmenopause: how far can assisted reproductive technology be trusted? PMID- 10674253 TI - Bleeding risk and reproductive capacity among patients with factor XIII deficiency: a case presentation and review of the literature. AB - Factor XIII deficiency is an uncommon, inherited bleeding disorder that usually manifests in infancy or early childhood, involving both boys and girls. We present the case of a woman who had experienced two previous intracranial bleeding events, and was treated before and during her current pregnancy with factor XIII concentrate. Her pregnancy was successful, and she experienced an uncomplicated vaginal delivery. To better understand the issues surrounding bleeding, reproductive capacity, and management of factor XIII deficiency during pregnancy, we conducted a systematic literature review using MEDLINE from 1966 to December 1998. We also examined the bibliographic references from all articles, and included all cases, case reports, or case series of patients with factor XIII deficiency. We retrieved data on 117 patients from 37 articles, the majority of which had type II deficiency. Among untreated patients with type II factor XIII deficiency, the literature suggests an elevated mortality rate due to uncontrolled bleeding and intracranial hemorrhage. Because of its high degree of efficacy, the evidence supports the use of life long prophylactic therapy with at least monthly infusions of factor XIII concentrate, including during pregnancy. The opinion that women with type II factor XIII deficiency have inevitable recurrent abortions, or that men are sterile, is not well substantiated. No data were found on whether treatment alters male reproductive capacity. A policy of universal factor XIII replacement, starting in childhood, will likely enable more patients to attain reproductive status. The development of an international data registry would optimally address both bleeding risk and reproductive capacity among patients with factor XIII deficiency. PMID- 10674254 TI - Gynecological and reproductive issues for women in space: a review. AB - Women have been an integral part of United States space crews since the initial flight of Dr. Sally Ride in 1983, and a total of 40 women have been selected as U.S. astronauts. This article examines the reproductive and gynecological aspects of selecting, training, medically certifying, and flying women in space. Gynecological data from the astronaut selection cycles in 1991 to 1997 are reviewed. In addition, the reproductive implications of delaying childbearing for a career as an astronaut and the impact of new technology such as assisted reproductive techniques are examined. The reproductive outcomes of U.S. female astronauts after spaceflight are also presented. Because women have gained considerable operational experience on the Shuttle and Mir, the unique operational considerations for preflight certification, menstruation control and hygiene, contraception, and urination are discussed. Medical and surgical implications for women on long-duration missions to remote locations are still evolving, and enabling technologies for health care delivery are being developed. There has been considerable progress in the development of zero-gravity surgical techniques, including laparoscopy, thoracoscopy, and laparotomy. The concepts of prevention of illness, conversion of surgical conditions to medically treatable conditions, and surgical intervention for long-duration spaceflights are explored in detail. There currently are no operational gynecological or reproductive constraints for women that would preclude their successful participation in the exploration of our nearby solar system. PMID- 10674255 TI - Obstetric implications of activated protein C resistance and factor V Leiden mutation. AB - An increasing number of reports have focused on activated protein C resistance (APCR) as it has been shown not only to be the most common genetic factor predisposing patients to thromboembolic disease but the most common identifiable cause overall. More than 90 percent of the cases of APCR are caused by the factor V Leiden mutation, in which a guanine to adenine substitution in the factor V gene at nucleotide position 1691 results in a glutamine to arginine switch at position 506. Recent studies have also pointed to evidence of an association between APCR/factor V Leiden mutation and hypertensive disorders of pregnancy, first and second trimester miscarriage, placental infarction, and placental abruption. PMID- 10674256 TI - [Role of the telophasic disc in the regulation of cytokinesis]. PMID- 10674257 TI - [Molecular study of a false outbreak of ticarcillin-resistant Pseudomonas aeruginosa infections in a clinical hematology department]. AB - Molecular methods for bacterial strain typing are becoming available outside teaching hospitals and large structures. Although it seems reasonable to use conventional markers, most notably the serotype, whenever possible, the limitations of these methods are particularly conspicuous with P. aeruginosa. Combined use of several methods such as quantitative antibiotic susceptibility testing and serotype determination has proved adequate for characterizing P. aeruginosa strains in some cases. In the case of the outbreak reported herein, this approach failed to provide high quality epidemiological data. In contrast, pulsed field gel electrophoresis ruled out epidemic spread of a P. aeruginosa strain in the clinical hematology department. PMID- 10674258 TI - [Inter-laboratory reproducibility of pulsed-field electrophoresis for the study of 12 types of Pseudomonas aeruginosa]. AB - The increasing hospital-to-hospital transmission of multiple drug-resistant bacteria is a major concern for bacteriology laboratories involved in nosocomial infection control. The interlaboratory reproducibility of pulsed-field gel electrophoresis (PFGE) for Pseudomonas aeruginosa typing was evaluated by asking four hospital laboratories (two in Lyon, one in Brest, and one in Marseille) to study 11 P. aeruginosa isolates, some of which were epidemiologically related, and the reference strain ATCC 27853. Two laboratories used the Genepath system, one the Chef DR II, system, and one the Chef Mapper system, Bio-Rad, restriction/Spe I. Profiles were read visually and by computerized comparison of restriction band molecular weights (Taxotron, software, PAD Grimont, Pasteur Institute, Paris, France). These two methods led to similar epidemiological conclusions. However, centralization of the data showed poor center-to-center reproducibility due to inadequate standardization of the procedure. PMID- 10674259 TI - [Pneumococcus observatory data in the Rhone-Alps region. Results from 1996]. AB - Throughout 1996, 22 hospital-based laboratories in the Rhone-Alpes region of France collected pneumococcal strains and used a standardized protocol to record the following data; patient age and sex; type of specimen; and determination of susceptibility to at least the following antibiotics: oxacillin 1 microgram and 5 micrograms, erythromycin (Ery), tetracycline (Tet), chloramphenicol (Chl), rifampin (Rmp), and loracarbef. For penicillin-nonsusceptible strains (PNSSs), which were identified based on results with oxacillin, MICs for penicillin G, amoxicillin (Amx), and cefotaxime (Ctx) were determined using the E Test, at the study site and agar dilution at the coordinating center. Of the 1153 strains, 65.5% were from adults and 31.8% from children; patient age was unknown in 2.7% of cases. PNSPs (MIC > 0.06 mg/l) contributed 32.9% of strains (I: 23.3%; R: 9.6%) and were more common in children (41.1%) than in adults (28.1%). The frequency of PNSSs varied across specimen types: 27.9% in blood cultures (305 strains), 15.6% in cerebrospinal fluid (32), 38.7% in protected bronchopulmonary specimens (31), 31.5% in unprotected bronchopulmonary specimens (434), 50.8% in acute otitis media (118), and 34.4% in other specimens (221). Among PNSSs, nonsusceptibility (I + R) to other antibiotics was variable: Ery, 62.1%; Tet, 41.5%; Chl, 40.4%; Rmp, 1.1%. Corresponding figures for the overall strain population were Ery, 33.3%; Tet, 22.7%; Chl, 22.8%; Rmp, 0.9%. In addition, 56.5% of PNSSs exhibited multiple drug resistance. Resistance to amoxicillin (MIC > 2 mg/l) was demonstrated for only 5 strains. No strains were resistant to loracarbef or cefotaxime. Serotypes of the 379 PNSSs were as follows: 23F, 26.6%; 14 (25.6%); 9V (18.2%), 6 (8.7%), 15 (5%), 19 (4.5%). PMID- 10674261 TI - [Surgical antibiotic prophylaxis: point evaluation of practices]. AB - Compliance of prophylactic antimicrobial therapy (PAMT) in surgical patients with consensus-based recommendations was evaluated at the Montfermeil Hospital Center, France, in 1996, based on data for given days. All patients who had surgery on the study days were included. Data on the patient, surgery, and PAMT were collected. Practices were evaluated based on seven criteria: need for PAMT, type of drug used, dosage, time of first administration in relation to the time the incision was made, time of administration during surgery, administration time schedule, and total duration. Of the 93 patients who had surgery on one of the five study days, 59.1% received PAMT. All seven evaluation criteria were met in 68.2% of cases. Failure to adhere to the recommended time of first administration was the most common form of noncompliance. PMID- 10674260 TI - [Bacterial contamination of multi-dose ocular solutions. A prospective study at the Grenoble Teaching Hospital]. AB - The bacterial contamination rate of multidose ocular solutions used by hospitalized patients was evaluated by culturing vial dropper tips and residual solution in vials. Bacterial colonies were counted and identified. Overall 39 (23.5%) selected vials were contaminated. Contamination rates were 17.7% (20/113) for vials used by ophthalmology ward patients and 35.8% (19/53) for vials used by internal medicine and gerontology patients (P < 0.02). The most commonly identified organisms were part of the normal commensal flora. Three ophthalmology patients were using vials contaminated with Pseudomonas aeruginosa. A significant (P < 0.01) positive correlation was found between vial contamination rate and duration of vial use. Vials containing an antimicrobial agent were less likely to be contaminated than vials without antimicrobials (P < 0.01). No clinical consequences of vial contamination were identified. However, ocular solution vial contamination carries a risk of infection. Our data are evidence of inadequate efficacy of preservatives present in ocular solutions. The standard practice of using ocular solution vials for seven days in health care facilities may need to be reappraised. Care should be taken to ensure that health care providers and patients understand the rules for ocular solution use. Unit-dose presentations may be preferable over multi-dose presentations for in hospital treatment. PMID- 10674262 TI - [Multidrug resistant bacteria in a psychiatric milieu]. AB - Isolation rates of multiple-drug resistant (MDR) bacteria were evaluated retrospectively in a psychiatric care facility. Over the six-year study period, 66 MDR bacterial strains were found. Methicillin-resistant Staphylococcus aureus contributed half of all MDR strains and 31% of all S. aureus strains. Among Pseudomonas aeruginosa strains, 22% were resistant to ticarcillin or imipenem, and among Enterobacteriaceae, 4.1% were MDR strains (production of a derepressed cephalosporinase or of an extended-spectrum beta-lactamase). Although most MDR strains were probably acquired during hospitalizations in short-term care facilities outside our institution, patient-to-patient transmission, either direct or via other individuals, cannot be ruled out. These data indicate that psychiatric care facilities should adopt the MDR strain monitoring strategies already used in other hospitals. PMID- 10674263 TI - [Genetic and molecular aspects of obesity: recent data]. AB - Obesity is a disease responsible for many serious complications. The sharp rise in the prevalence of obesity in many countries is supplying a powerful drive to basic and clinical research. Several genes responsible for monogenic murine obesity have recently been identified. One of these genes encodes the OB protein, or leptine, which is secreted by fat tissue and inhibits appetite by means of an effect on the hypothalamus. In humans, obese subjects carrying a mutation of this gene or of the leptine receptor have been identified. Several other genes implicated in human obesity have been mapped to chromosomes 1, 11, 18, and 20. Several transcription factors that control fat cell differentiation have been identified, such as C/ERB alpha, beta, and delta; ADD1/SREBP1, and PPAR gamma 2. It has been established that fat tissue can secrete many factors, including TNF alpha, CETP, IGF beta, TGF beta, PGE2, and LPA. Mitochondrial uncoupling proteins (UCPs) are recently characterized proteins capable of uncoupling respiration and contributing to energy expenditures. The hypothalamic neuropeptides and their receptors are a focus of active research. About ten of these neuropeptides have been identified. PMID- 10674264 TI - [Recent data from the literature on the biological and pathologic effects of electromagnetic radiation, radio waves and stray currents]. AB - Electromagnetic radiation is present in increasing amounts in our environment, and its potential effects on human (and animal) health has been investigated. It remains unclear whether the risk of acute childhood leukemia is associated with cumulative exposure to magnetic fields. An association with brain cancer and colon cancer has been suggested in electrical company workers. The radars used by police departments may increase the incidence of cancer. Electromagnetic radiation may play a role in a number of disorders such as depression and memory loss. It has been established that cell phones interfere with pacemakers only if direct contact occurs and have no effect if held in their normal position. Interferences have been reported between pacemakers and shop-lifting detectors. PMID- 10674265 TI - [A brief overview of the discovery of cell mortality and immortality and of its influence on concepts about aging and cancer]. AB - After having accomplished the miraculous performance that led us from conception to birth, then to sexual maturity and adulthood, natural selection failed to develop a more elementary mechanism capable of simply maintaining the results of this process forever. This failure is aging. Because few animals age in the wild, evolution could not give an advantage to animals with modifications due to aging. Natural selection benefits those animals that have the highest likelihood of effectively perpetuating their species because their vital systems have the larger reserve capacity they need to resist and survive predators, disease, injury, and extreme environmental conditions. Natural selection decreases after sexual maturity has been reached because at that stage the species would not derive additional advantages from individuals with larger physiological reserves. A species increases its likelihood of survival by investing its resources and energy into increasing its opportunities for fruitful reproduction rather than into prolonging its postreproductive life span. Most animals are mortal and undergo aging because investment of resources into keeping the body eternally youthful does not promote species survival as much as their investment into strategies that make reproduction more successful. PMID- 10674266 TI - Role of ras mutation in the progression of thyroid carcinoma of follicular epithelial origin. AB - The histological differentiation of thyroid carcinoma is known to correlate with prognosis. Ras oncogene mutations, which have been identified in various human cancers, have been suspected playing an important role in carcinogenesis and tumor progression. The purpose of this study was to clarify the mechanism of thyroid tumor progression, focusing on ras oncogenes. We examined ras mutations using nested polymerase chain reaction (PCR) and direct sequencing methods. The ras oncogene product was also examined immunohistochemically. Our results indicated that the incidence of ras mutations correlated with the histological differentiation of thyroid cancer. Three poorly differentiated carcinomas showed a higher rate of ras mutations than did 17 well-differentiated counterparts. Hot spots were not identified except for a relative accumulation of the N-ras gene at codon 61. There was a correlation between the immunoreactivity of the ras oncogene product and ras mutation, although the immunoreactivity of ras-p21 did not correlate with the histological differentiation. Mutation of the ras gene seemed to be one of the important events in the progression from well differentiated carcinoma to poorly differentiated thyroid carcinoma. PMID- 10674267 TI - Treatment with lansoprazole also induces hypertrophy of the parietal cells of the stomach. AB - The aim of the present study was to investigate whether not only omeprazole but also lansoprazole leads to hypertrophy of the parietal cells of the gastric mucosa. We investigated the gastric mucosa of 295 patients with gastro-esophageal reflux disease prior to therapy, after 8 weeks of treatment with 30 mg lansoprazole/day, and after 6 and 12 months of long-term treatment with 15 or 30 mg lansoprazole, or 20 mg omeprazole/day. In a small group of 48 patients, a follow-up examination was carried out 3 months after termination of the study. Following acute treatment with 30 mg lansoprazole/day, 84% of the patients revealed parietal cell hypertrophy, while under long-term treatment this figure rose to 89%. The grade of the parietal cell hypertrophy increased continuously under long-term treatment. With regard to the incidence and grading of parietal cell hypertrophy, no significant differences were found among the three groups receiving long-term treatment. Three months after termination of treatment with these proton-pump inhibitors, the hypertrophy of the parietal cells had largely disappeared again. This study shows that hypertrophy of the parietal cells in the gastric mucosa occurs not only under treatment with omeprazole, but also under therapy with lansoprazole, and is reversible when the proton pump inhibitors are discontinued. PMID- 10674268 TI - Expression of the plasminogen activator system and the inhibitors PAI-1 and PAI-2 in posttraumatic lesions of the CNS and brain injuries following dramatic circulatory arrests: an immunohistochemical study. AB - Plasminogen activators as inducible extracellular serine proteases are involved in a variety of processes, such as the degradation of brain structures. In regions of brain degradation, an increase in the expression of genes encoding cytokines and proteinases has recently been demonstrated. We tested the hypothesis, whether the plasminogen activator system as well as the plasminogen activator inhibitors are expressed and possibly involved in a proteolytic cascade that breaks down the extracellular matrix as a result of ischemic or posttraumatic brain destructions. To study this supposition, we investigated immunohistochemically the expression of tPA, uPA and its receptor, the plasminogen activator inhibitors PAI-1 and PAI-2, tetranectin as well as the laminin breakdown as an event of secondary brain injury. Brain tissue from 21 autopsy cases with severe brain injuries, material from 14 ischemic infarcts and 11 controls with acute hypoxia were used. All components of the plasminogen activator system studied were over-expressed immunohistochemically in reactive astrocytes, microglia and endothelial cells around the lesion zone. Tetranectin showed an analogous distribution to the plasminogen activator system. A reduced immunoreactivity of laminin within the identical region of destruction was detected concomitant with laminin remnants in perivascular macrophages, so that a remarkable role of the plasmin cascade in the degradation of extracellular matrix proteins in the brain is taken into consideration. PMID- 10674269 TI - Time-dependent changes of decorin in the infarct zone after experimentally induced myocardial infarction in rats: comparison with biglycan. AB - Decorin, a small dermatan sulphate proteoglycan, has been postulated to interact with other components of the extracellular matrix. We examined time-dependent changes of decorin in the infarct zone after experimentally induced myocardial infarction in rats by Northern blotting, in situ hybridization, and immunohistochemistry. The expression of decorin mRNA was compared to that of biglycan mRNA. Northern blotting demonstrated that the decorin mRNA expression was not increased in the infarct zone on day 2, while increased biglycan mRNA was observed at that time (average 3.1-fold increase). Decorin mRNA expression was increased on day 7, and reached a peak (average 2.2-fold increase) around day 14. Biglycan mRNA expression also reached a peak level around day 14 (average 13.3 fold increase). In situ hybridization revealed that mRNA signals for decorin did not appear in the infarct zone on day 2, while biglycan mRNA signals were observed. Decorin mRNA signals were observed in spindle-shaped mesenchymal cells in the infarct peripheral zone on day 7. The decorin mRNA signals appeared later than those of biglycan. Immunopositive staining for decorin was observed in the infarct zone on day 7. The present results demonstrated a time-dependent increase in decorin mRNA expression in mesenchymal cells in the infarct zone in rats. Decorin mRNA appeared later and was increased to a lower extent in the infarct zone than biglycan mRNA. PMID- 10674270 TI - Importance of estrogen receptors for the behavior of invasive micropapillary carcinoma of the breast. Review of 68 cases with follow-up of 54. AB - The aggressiveness of invasive micropapillary carcinomas of the breast (MPCa) is still controversially discussed. Therefore, we investigated a total of 68 cases and studied the evolution of 54. MPCa were frequently well-differentiated cancers with the following positivities for immunohistochemistry: 74.5% estrogen receptor (ER+), 46.3% progesterone receptor (PR+), 66% Bcl2+, and 36.4% C-erbB-2+. However, in 90.5% of the cases lymph nodes were involved at diagnosis, and 70.6% of T1 tumors showed wide metastatic spread. After a mean follow-up of 52.6 months, 55.6% of the patients were disease free (DF), 7.4% had disseminated disease and 37% had died. Univariate analysis showed significant differences. Thus, the DF group of patients included 90% of those having tumors with an associated colloid pattern, 73.3% of the ER positive tumors, none of the C-erbB-2 positive tumors, and 100% of the tumors with no axillary metastasis, 77.8% of those with metastasis to up to 3 nodes, and 47.2% of those metastasizing to 4 or more nodes. However, using Cox's regression model for survival analysis, ER was the only factor associated with duration of survival (p = 0.0175). In conclusion, although long-term survival in MPCa is determined by involvement of lymph nodes, as is the case in any other breast cancer type, their short-term evolution is influenced by other factors, mostly by estrogen receptors. PMID- 10674271 TI - RB protein expression in human endometrial carcinomas--an immunohistochemical study. AB - The aim of the current study was to investigate the immunohistochemical expression of the retinoblastoma protein (pRB) in formalin-fixed, paraffin embedded specimens obtained from 62 patients suffering from endometrial cancer. The avidin-biotin-peroxidase detection system with microwave pretreatment and the mouse anti-human NCL-RB1 monoclonal antibody were used. Heterogeneous nuclear immunostaining for the pRB was generally observed in the glandular cells in 59 out of 62 (95%) endometrial carcinomas, while stromal components were unreactive. In one case of stage Ic endometrioid adenocarcinoma, a small percentage of glandular cells (5%) stained positively with the anti-RB antibody, while two other tumors (stage IIa adenosquamous carcinoma and stage IIIa endometrioid adenocarcinoma) were pRB negative. In the cases with concomitant hyperplastic and neoplastic endometrial lesions, pRB immunoreaction was heterogeneous in the hyperplastic endometrial cells and in the adjacent neoplastic endometrium. Moreover, eight cases of endometrial carcinoma harboring K-ras codon 12 gene point mutation overexpressed pRB (more than 80% of glandular endometrial cells were positive) immunohistochemically, while none of three pRB negative slides had a K-ras gene alteration. Our data support the view that the pRB is expressed in most of the human endometrial neoplasms, but the lack of pRB immunoreactivity may correspond with the retinoblastoma gene rearrangements in a subset of advanced endometrial carcinomas. PMID- 10674272 TI - Concentration of iron and distribution of iron and transferrin after experimental iron overload in rat tissues in vivo: study of the liver, the spleen, the central nervous system and other organs. AB - The purpose of this study was to estimate the iron concentration in the liver, spleen and brain of control rats and rats overloaded with iron and to determine the distribution of iron and of transferrin (TF). Iron was administered to Wistar rats by food supplemented with 3% carbonyl iron for 3 months, or intraperitoneally, or intraveneously as iron polymaltose for 4 months (total administered dose: 300 or 350 mg/rat, respectively). Iron concentration was estimated by atomic absorption spectrophotometry and iron- and TF-distribution histochemically and immunohistochemically, respectively. In control rats the organ with the highest iron content was the spleen, followed by the liver and brain. After iron loading the increase of iron in the liver was greater than that of the spleen; iron concentration in the brain did not change significantly. Distribution of iron in the liver was in Kupffer cells throughout the lobule and in hepatocytes at its periphery. No difference in the number of positive cells or staining intensity for TF was observed between control rats and iron overloaded animals in the liver or central nervous system (CNS); the spleen was negative for TF. Distribution of TF in the liver showed a centrilobular localisation in hepatocytes. TF reaction in the brain occurred in oligodendrocytes, vessel walls, choroid plexus epithelial cells and some neurons. In conclusion, experimental iron overload in rats leads to iron uptake mainly by reticuloendothelial (RE) cells and hepatocytes, indicating that hepatocytes are of particular importance for iron metabolism. Iron uptake by the brain was not significant, probably because the brain is protected against iron overload. Iron overload did not influence location and quantity of TF in the liver and CNS, whereas the visualisation of iron and TF did not coincide. This indicates that TF may have other functions beyond iron transport. PMID- 10674273 TI - Sustentacular cells in sporadic paraganglioma-like medullary thyroid carcinoma: report of a case with diagnostic and histogenetic considerations. AB - Sustentacular cells (SCs) are glial supporting cells of the fetal, adult normal, and neoplastic extra and adrenal human chromaffin cell lineage. SCs have also been identified in some cases of medullary thyroid carcinoma (MTC), raising both diagnostic and histogenetic problems. We report a rare case of a sporadic paraganglioma-like variant of MTC showing numerous S-100 protein and glial fibrillar acid protein (GFAP) positive SCs encircling neoplastic cells in a nesting (Zellballen) pattern similar to that observed in paragangliomas or pheochromocytomas. Although stromal amyloid deposits were only focally detected, diagnosis was immunohistochemically confirmed by immunoreactivity of the neoplastic cells for cytokeratin, CEA, calcitonin, chromogranin A, neuron specific-enolase, and synapthopysin. As for the histogenesis of SCs, if we assume that MTC is a neural crest-derived tumor, it is likely that these cells reflect the ability of the common precursor cell to differentiate towards a sustentacular type glial cell lineage in addition to the typical neuroendocrine one. This viewpoint is supported by the evidence that rare cases of MTC may contain neoplastic or supporting cells showing a multidirectional differentiation (usually neuroendocrine and melanocytic) that recapitulates the different cell lineages arising from the developing neural crests. PMID- 10674275 TI - Spindle cell rhabdomyosarcoma in adults. A case report and literature review. AB - Spindle cell rhabdomyosarcoma, a recently described variant of embryonal rhabdomyosarcoma in children, carries a favorable prognosis when compared with other types of rhabdomyosarcoma. This tumor is rare in adults, and only four cases have been documented previously. The clinicopathological study of such a case occurring in the retroperitoneal space of a 53-year-old man is herein reported. The patient died of uncontrolled local recurrence and hepatic metastases seven months after diagnosis. Based on the analysis of the data of the five cases reported, including the present one, it can be stated that spindle cell rhabdomyosarcoma in adults is not associated with the favorable outcome observed in the pediatric population. PMID- 10674274 TI - Foreign-body reaction to silastic burr-hole covers with seroma formation: case report and review of the literature. AB - Because silastic material is one of the most commonly used biomaterials in modern medicine, the biocompatibility of these implants is still a source of long standing controversy. Though several studies have established silastic material as biologically inert, numerous authors have repeatedly described characteristic pathological tissue responses to silicone and its elastomeres. We report a case of foreign-body reaction to silastic burr-hole cover with successive formation of a seroma following resection of an olfactory groove meningioma. Within 30 days postoperatively, the patient developed a marked bulge in the glabbelar region. Histopathological examination revealed a seroma-like lesion obviously caused by a chronic inflammatory allergic reaction to the silastic burr-hole cover. Although the silicone-induced tissue damage clinically shows a wide variability and a conclusive model of pathogenesis is presently not available, the histopathological findings in some patients, in the form of granulomatous lesions and inflammatory cell response, might partly be due to an immunological reaction. Such a reaction has been previously described both clinically and experimentally, as detected in our patient. In addition, a review of the literature is given. PMID- 10674276 TI - Interaction of desipramine with steroid hormones on experimental anxiety. AB - The present study analyzes if estradiol benzoate and/or progesterone interact with desmethylimipramine (DMI) to diminish experimental anxiety. The animal model of anxiety used was the conditioned defensive burying test. Dose response curves for DMI (0.625, 1.25 and 2.5 mg/kg, every 24 h, during 21 days), estradiol benzoate (0.5, 1.0, 2.0 and 4.0 micrograms/rat, 48 h) and progesterone (0.5, 1.0 and 2.0 mg/rat, -4 h) were made in ovariectomized rats. DMI per se decreased dose dependently the cumulative burying time, an effect considered as anxiolytic-like. Progesterone produced a decrease in burying at the highest dose, while estradiol benzoate had no effect on defensive burying. Both, progesterone (0.5 mg/rat) and estradiol benzoate (4.0 micrograms/rat) were able to decrease the cumulative burying behavior when injected with a subthreshold dose of DMI (1.25 mg/kg). In addition, the effect of DMI (1.25 mg/kg) plus the combination of estradiol benzoate and progesterone, sequentially administered (48 h and 4 h before the tests, respectively), also produced a synergistic decrease in burying behavior. In general, the treatments produced no changes in burying behavior latency, neither in spontaneous ambulation or in nociception. It is concluded that DMI synergizes its anxiolytic-like effect when administered with estradiol alone or in combination with progesterone. Present data provide experimental evidence suggesting an interaction between hormones and antidepressants. Results are discussed on the basis of the interaction between steroids and serotonergic or GABAergic receptors. PMID- 10674277 TI - Immune activation in the early puerperium is related to postpartum anxiety and depressive symptoms. AB - The pathophysiology of the postpartum blues, common transient mood disorders in the first week postpartum, has remained elusive. Recently, however, it has been shown that depression and anxiety disorders are accompanied by activation of the inflammatory response system (IRS). This study was developed to determine whether the postnatal blues is associated with IRS activation. Serum concentrations of interleukin-6 (IL-6), IL-6 receptor (IL-6R), gp130 (the IL-6 signaling protein), IL-1R antagonist (IL-1RA) and leukemia inhibitory factor receptor (LIFR) were assayed in 22 nonpregnant women and in 91 pregnant women before delivery and 1 and 3 days after delivery. On each occasion the parturient women completed the State version of the Spielberger State-Trait-Anxiety-Inventory (STAI) and the Zung Depression Rating Scale (ZDS). Serum IL-6, IL-1RA and LIFR were significantly higher in pregnant women at the end of term than in nonpregnant women. PMID- 10674278 TI - Neurohormonal responses to D-fenfluramine in healthy elderly subjects. A placebo controlled study. AB - Considering age-related changes in serotonin (5HT) function, we examined normative data of prolactin (PRL) and cortisol (CORT) responses to D-fenfluramine (D-FEN) in healthy elderly subjects. Twenty-three healthy male and female volunteers aged 60-86 participated in a single-blind, placebo-controlled, fixed order, crossover-design challenge test. Two baseline PRL and CORT values and the responses of these hormones to 30 mg of oral D-FEN and placebo over a 4 h period were measured on two separate sessions. PRL and CORT responses were significantly greater following D-FEN than after placebo. Peak PRL responses (maximum change from baseline following D-FEN) were relatively robust compared to peak CORT responses. Peak PRL concentration was positively correlated with plasma D-nor-FEN concentration. Gender and aging had no effect on hormonal responses in the elderly. Although the weight adjusted dose used in this study was higher than the therapeutic dose of D-FEN, PRL responses were modest and only two participants experienced side effects. D-FEN is a safe serotonergic probe and PRL responsivity to D-FEN is a reliable index of central 5HT function in the elderly. An age related decline in serotonergic function must be considered in determining the dose requirement for maximal hormonal responses to D-FEN challenge tests in the elderly. PMID- 10674279 TI - Dexamethasone suppression of corticosteroid secretion: evaluation of the site of action by receptor measures and functional studies. AB - A dose of dexamethasone was determined in rats (50 micrograms/kg s.c.) that suppressed the corticosterone response to restraint stress by 80%. Corticosteroid receptor occupancy estimates found that the 50 micrograms/kg s.c. dose of dexamethasone had no significant effect on available glucocorticoid receptor (GR) or mineralocorticoid receptor (MR) binding in brain regions (hypothalamus, hippocampus and cortex); on the other hand dexamethasone produced a selective and significant decrease in available GR in peripheral tissues (pituitary and spleen). Functional studies showed that the 50 micrograms/kg s.c. dose of dexamethasone completely blocked the effects of corticotropin-releasing hormone (CRH; 0.3-3.0 micrograms/kg i.p.) on corticosterone secretion, but did not inhibit the corticosterone response to an adrenocorticotropin hormone (ACTH; 2.5 I.U./kg i.p.) challenge. These studies indicate that this dose of dexamethasone exerts its inhibitory effects on the HPA axis primarily by acting at GR in the pituitary. The plasma dexamethasone levels produced by this dose of dexamethasone are similar to those present in humans the afternoon after an oral dexamethasone suppression test (DST), a time at which many depressed patients escape from dexamethasone suppression. These results support and extend other studies which suggest that the DST provides a direct test of the effects of increased GR activation in the pituitary on ACTH and cortisol secretion. PMID- 10674280 TI - Hormonal markers of stress response following interruption of selective serotonin reuptake inhibitor treatment. AB - Depressive illness is associated with loss of the usual regulation of stress responsive hormonal and neurotransmitter systems, and antidepressants have intrinsic effects reducing the activity of these systems. Abrupt interruption of treatment with some antidepressants has been associated with a self-limited syndrome of physical and psychological symptoms distinct from relapse, of which drug half-life appears to be the major determinant. We hypothesized that reactivation of stress-response systems could play a role in this syndrome and studied the effects of treatment interruption in patients successfully treated with the antidepressant fluoxetine, paroxetine or sertraline. During placebo substitution, interruption of paroxetine was associated with statistically significant increases in plasma IGF-1 and heart rate. These data suggest that some activation of physiologic stress-responses may accompany symptom increases during treatment interruption of shorter half-life agents. PMID- 10674281 TI - Muscarinic cholinergic mediation of the GH response to gamma-hydroxybutyric acid: neuroendocrine evidence in normal and parkinsonian subjects. AB - We have recently reported that parkinsonian patients show a significant GH response to gamma-hydroxybutyric acid (GHB), but not to gamma-aminobutyric acid (GABA)-ergic drug administration. In order to establish whether muscarinic cholinergic receptors mediate the GH secretion induced by GHB, normal men and parkinsonian patients were tested with GHB both in the absence and in the presence of the anticholinergic agent, pirenzepine. Both normal controls and parkinsonian patients showed a significant serum GH rise in response to GHB (25 mg/kg body weight p.o.) even though a slightly, but significantly lower response was observed in parkinsonian patients. Pretreatment with pirenzepine (100 mg p.o. 2 h before GHB) completely suppressed the GHB-induced GH release in both normal controls and parkinsonian patients. These data indicate that a cholinergic mechanism mediates the GH response to GHB in normal men. In addition the data indicate that this pathway is preserved in the parkinsonian brain. PMID- 10674282 TI - Neuroendocrine response to film-induced sexual arousal in men and women. AB - The psychoneuroendocrine responses to sexual arousal have not been clearly established in humans. However, we have demonstrated previously that masturbation induced orgasm stimulates cardiovascular activity and induces increases in catecholamines and prolactin in blood of both males and females. We presently investigated the role of orgasm in producing these effects. Therefore, in this study parallel analysis of prolactin, adrenaline, noradrenaline, and cortisol concentrations, together with cardiovascular variables of systolic/diastolic blood pressure and heart rate were undertaken during film-induced sexual arousal in nine healthy adult men and nine healthy adult women. Blood was drawn continuously via an indwelling cannula and connected tubing system passed through a mini-pump. In parallel, the cardiovascular parameters were recorded continuously via a computerised finger-cuff sensor. Subjective sexual arousal increased significantly in both men and women during the erotic film, with sexual arousal eliciting an increase in blood pressure in both males and females, and plasma noradrenaline in females only. In contrast, adrenaline, cortisol and prolactin levels were unaffected by sexual arousal. These data further consolidate the role of sympathetic activation in sexual arousal processes. Furthermore, they demonstrate that increases in plasma prolactin during sexual stimulation are orgasm-dependent, suggesting that prolactin may regulate a negative-feedback sexual-satiation mechanism. PMID- 10674283 TI - Long-term residual complaints and psychosocial sequelae after remission of hyperthyroidism. AB - The issue of residual complaints after treatment for hyperthyroidism in current euthyroid patients was investigated by means of a survey. Patients treated for hyperthyroidism were selected from medical records of the previous 6 years in two Dutch University Clinics. After the exclusion of patients with comorbidity, 303 one-time hyperthyroid respondents were included in the analysis. A total of 77% of these patients had been diagnosed with Graves' Disease. The newly developed Hyperthyroidism Complaint Questionnaire (HCQ), was used to quantify problems of somatic and mental functioning. The SymptomsCheckList-90 (SCL-90) was used to assess self-reported psychopathological symptoms, the Nottingham Health Profile was used to measure perceived health/quality of life. Dysthyroid patients (n = 20) had a mean HCQ-score of 14.5 (+/- 8.1) complaints; patients who reported euthyroidism for less than 12 months (n = 171) had a mean of 9.3 (+/- 7.6) residual complaints; patients who reported euthyroidism for more than 12 months (n = 54) a mean of 6.6 (+/- 6.8) residual complaints. On each dimension of psychopathology covered by the SCL-90, including depression and anxiety, approximately one third of the total sample had a score exceeding 80% of adult females. According to the NHP lack of energy was evident in 53% of all respondents. Over one third of patients with a full-time job were unable to resume the same work after treatment. It appears that many of these patients are in need of psychological support. PMID- 10674284 TI - 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX): toxicological properties and risk assessment in drinking water. AB - MX (3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone), one of the byproducts formed during the chlorine disinfection process of drinking water, shows strong mutagenic activity for Salmonella strains in the Ames test. In several countries, the contribution of MX to the total mutagenicity of drinking water is estimated to range from 7% to 67%. To assess the risk of MX for human health, we summarized the toxicological properties of MX and estimated the tolerable daily intake (TDI) or tolerable concentration in drinking water. MX is genotoxic in cultured mammalian cells and causes in vivo DNA damage in several tissues. MX is carcinogenic for rodents in addition to possessing skin and gastric promotion activities. From these toxicological profiles of MX, we estimated the virtual safety dose (VSD) for genotoxic action as 5 ng/kg/d and the TDI for non-genotoxic action of MX as 40 ng/kg/d. We assumed a tolerable MX concentration of 150 ng/L in drinking water. Because of the uncertainty about human genotoxicity, however, and the lack of information on reproductive or developmental toxicity, the estimated tolerable dose level may be provisional. PMID- 10674285 TI - Particulate air pollution and asthma: a review of epidemiological and biological studies. AB - The link between exposure to air pollution and exacerbation of asthma symptoms has been investigated by epidemiological study and by direct biological experimentation. In asthmatics, epidemiological studies generally show a positive correlation between the particulate fraction of air pollution and increased morbidity, although roles for other co-pollutants (for example, ozone) are implicated as well. Direct experimentation using air pollutants, especially particles, to investigate their effects on humans or on animal models of asthma provides corroboration of the epidemiology and has begun to identify the pathophysiological mechanisms involved. We begin this review with an overview of air pollution, followed by a survey of the epidemiological and experimental data regarding air pollution particles and asthma. We finish with a discussion of directions for future research. PMID- 10674286 TI - Inverse social gradient of secondary immune response parameters in children. AB - BACKGROUND: In children, the prevalence of allergy increases with increasing socioeconomic status. If frequent immune response stimulation by infections protects against development of allergic diseases, then a social gradient in infections should exist. The aim of our study was to assess the relation between social class, immune parameters, and the prevalence of respiratory infections in children. METHODS: A cross-sectional survey examined children aged 5 to 14 years in 3 communities of Sachsen-Anhalt, Germany. Data of 1724 children were gathered by a parent-completed questionnaire and analyses of blood samples. Social class was defined by parental educational level. Immune parameters included serum immunoglobulin (Ig)G, IgA, and IgM; the C3c component of complement; and the total leukocyte count. The period prevalence of febrile colds and lifetime prevalences of physician-diagnosed bronchitis, tonsillitis, otitis media, and pneumonia were assessed from parents' reports. Adjusted odds ratios for the association between social class and belonging to the group of children with immune parameter levels in the upper 50th or 75th percentile or experiencing respiratory infections were calculated by logistic regression. RESULTS: Social class was inversely associated with secondary immune response parameters (IgG, IgA), whereas indicators of primary immune reactions and inflammation (IgM, C3c, leukocytes) were not related to social status. While an inverse social gradient was found for the period prevalence of febrile colds, the frequency of bronchitis, tonsillitis, and otitis media decreased with decreasing social class. CONCLUSIONS: Health inequalities exist in immune reactions and respiratory infections in children from different social classes. We hypothesize that in children from lower social classes, increased frequency of infections stimulated the secondary immune response and protected against more severe courses of respiratory infections. PMID- 10674287 TI - Atmospheric carcinogens in Rio de Janeiro during the summer of 1998/99: benzo[a]pyrene and benzene. AB - Rio de Janeiro, the second largest city in Brazil, is affected by severe pollution episodes and presents a high respiratory cancer incidence in comparison with the rest of the country. To monitor atmospheric pollution during the summer of 1998/1999 and to estimate the impact of organic pollution on public health, we determined the levels of two carcinogenic organic chemicals, benzo[a]pyrene and benzene, in four distinct sites throughout the city. A review of the levels recorded in other urban areas worldwide during the last ten years indicates that the benzo[a]pyrene (< or = 0.70 ng/m3) and benzene (< or = 11 micrograms/m3) concentrations found in Rio are relatively low. The highest levels were generally recorded in developing Asian countries, whereas the lowest values were found in North America. Unlike urban areas in temperate zones, pollution derived from domestic heating is minor in Rio de Janeiro, where most of the benzo[a]pyrene and benzene pollution originates from vehicular traffic. The quite distinct fuels used in light-duty vehicles in Brazil, combined with strong light incidence and increased rainfall during the summer, also contribute to diminish the levels of such pollutants. PMID- 10674288 TI - Indoor allergen exposure in west and East Germany: a cause for different prevalences of asthma and atopy? AB - West and East Germans have been living in two different political systems for 40 years. These two populations have become a classic epidemiological example for the hypothesis that lifestyle changes accompanying the industrial and economic development of modern societies are responsible for an increase in the prevalence of atopic diseases. A higher prevalence of atopic sensitization, asthma, and hay fever was found in young West Germans after the unification. It has been hypothesized that this phenomenon was at least partially due to the installation of insulating windows and central heating systems in Western homes, favoring the growth of microorganisms like mites and moulds and increasing indoor allergen exposure. This review summarizes studies that have investigated reservoir concentrations of indoor allergens in public buildings and private homes in East and West Germany. Whereas a higher prevalence of atopic sensitization in West Germans was found for nearly all tested allergens (cat, mite, pollen), allergen exposure was higher only for cat allergens, but probably not for mite and cockroach allergens or moulds. The published data do not support the view that the differences in specific sensitization are caused by differences in the exposure to specific allergens. PMID- 10674289 TI - Complex interconvertibility of nitrogen oxides (NOX): impact on occupational and environmental health. AB - Oxides of nitrogen have been implicated in a vast number of environmental and occupational health effects, some of which lack concrete mechanisms. Whereas certain compelling pieces of evidence link a particular nitrogen oxide to a particular adverse health effect, other reports seem to implicate virtually all the oxides in one form of toxic process or the other. Such diversity has probably emerged because each oxide of nitrogen possesses a different oxidation state, in which each form exerts different important levels of biological activity. Most important, each nitrogen oxide readily interconverts into another oxide at a very fast rate. This property of rapid interconvertibility poses problems to researchers in identifying with great confidence the actual oxide of nitrogen that is responsible for a specific occupational and environmental health effect. This paper reviews the complex nature of the nitrogen oxides (represented collectively as NOX) to explore the extent to which their acute or chronic exposure could be associated with toxicity. The nomenclature of the nitrogen oxides is outlined as a necessity for clarity and simplicity in understanding their reactions and interconvertibility and how they affect health. The natural occurrence and sources of occupational and environmental exposures and effects are critically evaluated and analyzed. PMID- 10674290 TI - Use of reversed-phase high-performance liquid chromatography in QSAR analysis of 2,4-dihydroxythiobenzanilide analogues. AB - Thiobenzanilides are found to show strong biological activity as antimicrobial, antimycotic, and tuberculostatic agents. In addition, they are relatively weakly toxic to higher organisms. A large set of new (N-phenyl-)-2,4 dihydroxybenzenecarbothioamide derivatives was obtained. Preliminary studies showed high microbiological action of some of them. In the process of chromatographic analysis, several different chromatographic parameters were obtained. In case of RP-HPLC, these parameters correspond to hydrophobicity of the solute. Obtained chromatographic parameters exhibited moderate correlation with calculated log P parameter. Linear dependence of bacteriostatic or fungostatic activity on lipophilicity was observed. The degree of correlation of different parameters was compared. The lipophilicity of analysed tioamides was the most important factor responsible for fungostatic and bacteriostatic activity. In comparison to methanol eluent system, chromatographic parameters obtained in acetonitrile system were better correlated with bioactivity. Conversely with the calculated log P values, the experimentally derived parameters exhibited significant higher correlation to fungostatic activity determined on dermatophytes. While in case of other tested microorganisms log P was comparably or sometimes slightly better correlated. PMID- 10674291 TI - Chemical diversity approach for evaluating mechanistic relatedness among toxicological phenomena. AB - The CASE/MULTICASE structure-activity relationship (SAR) system was used to assess a new procedure to investigate the mechanistic relatedness of various toxicological endpoints. The method consisted of predicting the activity of 10,000 randomly selected chemicals using validated and characterized SAR models from a variety of biological and toxicological endpoints. The prevalence of chemicals predicted to possess the ability to induce two or more toxicological effects simultaneously should provide a measure of the mechanistic relatedness of these phenomena. Eight toxicological endpoints were predicted and the results were compared to predictions based on an eye irritation SAR model. Allergic contact dermatitis demonstrated a 29.6% greater than expected overlap between expected and observed results (p < 0.001). Similar results were seen for respiratory hypersensitivity (33.1%), sensory irritation (28.9%), cell toxicity (25.9%), and Ah receptor binding (19.8%). A lesser degree of overlap was seen between eye irritation and Salmonella mutagenicity (11.5%) and the inhibition of gap junction intercellular communication (6.7%). Moreover, a negative overlap, suggesting possibly an antagonistic phenomena, was observed between eye irritation and alpha 2 mu-induced nephropathy. These results indicate that this method can provide a useful tool to investigate mechanistic relatedness between diverse toxicological endpoints. PMID- 10674292 TI - Using molecular quantum similarity measures as descriptors in quantitative structure-toxicity relationships. AB - In this paper molecular quantum similarity measures (MQSM) are used to describe molecular toxicity and to construct Quantitative Structure-Toxicity Relationships (QSTR) models. This study continues the recently described relationships between MQSM and log P values, which permits to use the theoretical MQSM as an alternative to the empirical hydrophobic parameter in QSPR studies. In addition a new type of MQSM is presented in this work: it is based on the expectation value of electron-electron repulsion energy. The molecular properties studied here, as application examples are aquatic toxicity, toxicology on Bacteria and inhibition of a macromolecule employing four different molecular sets. PMID- 10674293 TI - QSAR analyses of the toxicity of aliphatic carboxylic acids and salts to Tetrahymena pyriformis. AB - Carboxylic acids have been conspicuously absent from the quantitative structure activity relationship (QSAR) literature. This study investigated the aquatic toxicity (log(IGC50(-1)) of selected mono- and di-carboxylic acids and their sodium, or disodium salts, tested in the Tetrahymena population growth assay. The relationship between log(IGC50(-1)) and hydrophobicity as described by the 1 octanol/water partition coefficient (log Kow) revealed a distinct sub-class. The relationship [log(IGC50(-1)) = 0.27(log Kow) - 0.68; n = 16, r2 = 0.943, s = 0.07, F = 233, Pr > F = 0.0001] was derived for mono-carboxylic acids. The QSAR [log(IGC50(-1)) = 0.19(log Kow) - 0.66; n = 9, r2 = 0.951, s = 0.08, F = 135, Pr > F = 0.0001] was generated for the di-carboxylic acids. Regression analysis of data for the monosodium carboxylic acid salts yielded the model, log(IGC50(-1)) = 0.60 (log Kow) + 0.58; n = 4, r2 = 0.932, s = 0.19, F = 41.2, Pr > F = 0.008. Values for the ionization constant (pKa) and the energy of the lowest unoccupied molecular orbital (ELUMO) do not vary within the sub-class for saturated acids. Moreover, pKa and ELUMO did not describe differences in toxicity between sub classes of saturated aliphatic carboxylic acids and salts. However, these descriptors did vary for unsaturated acids. Inclusion of unsaturated acids afforded the derivation of a global response-surface for all aliphatic carboxylic acids, log(IGC50(-1)) = 0.25(log Kow) - 0.13(ELUMO) - 0.54; n = 34, r2 = 0.850, s = 0.138, F = 87.9, Pr > F = 0.0001. Outliers to the response-surface included small molecules that provided 2-positions in which the molecule could potentially undergo electrophilic attacked and other more large, hydrophobic molecules. PMID- 10674294 TI - Should research become mandatory in future vocational training of general practitioners? PMID- 10674295 TI - From teaching to learning. Experiences of small CME group work in general practice in Sweden. AB - OBJECTIVE: To describe the experiences of learning in small groups and the impact of small group continuing medical education (CME). DESIGN: Literature review, personal communication and critical reflection. SETTING: General practice in Sweden. SUBJECTS: Small CME groups. MAIN OUTCOME MEASURES: Occurrence, themes and impact of small CME groups. RESULTS: In 1998, there were approximately 230 small CME groups in Sweden, which means that nearly half of Swedish general practitioners (GPs) participated in such activities. Although widely used in Sweden, small CME groups are less practised than "traditional" CME activities, such as lectures. Group work might enhance knowledge development, enable the assessment of individual learning needs and facilitate the adoption of national guidelines and agreements between primary and secondary care. A competent group leader is crucial. CONCLUSION: A transition from passive to interactive learning in small groups is recommended. PMID- 10674296 TI - The influence of sociodemographic characteristics on well-being and symptoms in a Swedish community. Results from a postal questionnaire survey. AB - STUDY OBJECTIVE: To assess the influence of sociodemographic characteristics on self-reported well-being and symptoms. DESIGN: A postal questionnaire was sent to a representative population sample drawn from the population census. SETTING: The municipality of Habo, Sweden. PARTICIPANTS: Out of 1312 subjects in the population sample, 827 (63%) participated in the study, i.e. answered the questionnaire. RESULTS: Sociodemographic characteristics significantly influenced most well-being variables and symptoms. The prevalence of symptoms in the categories depression and tension, as well as headache, decreased while most other symptoms increased with age. Women had more symptoms than men. Married subjects, compared to others, had higher social and mental but lower physical well-being. Subjects from households with up to three persons, and subjects with comprehensive school only, had lower physical well-being than other subjects. Working subjects generally had a higher well-being than non-working subjects. CONCLUSION: Sociodemographic characteristics had a significant influence on most well-being variables and symptoms. PMID- 10674297 TI - Detecting child sexual abuse in general practice: a retrospective case-control study from Wales. AB - OBJECTIVE: To investigate if routine medical contacts provide indicators that would assist general practitioners in detecting male child abuse. DESIGN: A case control study of the general practice records of male victims of a major episode of school-based child abuse and matched controls. SETTING: General practices serving cases and controls from two South Wales schools. SUBJECTS: 107 abused boys and 107 aged-matched controls. RESULTS: No boys had disclosed sexual abuse to general practitioners. Somatic and behavioural symptoms were reported by small numbers in both groups (18 cases/25 controls). Odds ratios showed no significant positive association between abuse and numbers of boys presenting with symptoms (odds ratio 0.66; 95% confidence interval 0.32-1.37), and no difference could be found in the nature of the symptoms complained of by boys from the two groups. However, abused boys were more likely than controls to present with symptoms that persisted for more than a year (eight cases compared with one control). The two tailed p-value calculated using the Fisher exact test was 0.035, suggesting a statistically significant association between abuse and persistent symptoms. CONCLUSIONS: Sexually-abused boys are unlikely to visit general practitioners with open requests for help, and do not appear to present with behavioural or somatic symptoms different from those presented by non-abused boys. However, where boys complain of persistent, inexplicable, somatic or behaviour problems over a period of time, the possibility of abuse should be considered. PMID- 10674298 TI - The management of chronic neck pain in general practice. A retrospective study. AB - OBJECTIVE: To describe the management in patients with chronic non-specific neck pain in general practice. DESIGN: A descriptive, questionnaire-based retrospective study. SETTING: General practices in the Netherlands. PATIENTS: 517 patients with chronic non-specific neck pain. MAIN OUTCOME MEASURES: Nature and frequency of diagnostic procedures, therapeutic interventions and referrals by the general practitioner (GP). RESULTS: Forty-four per cent visited the GP for neck pain in the previous year. Of the patients who did visit the GP in the previous year, 32% did not receive a diagnostic modality, 31% did not receive therapy and 43% were not referred. The most frequently applied diagnostic and therapeutic modalities were physical examination (66%) and pain medication (58%), respectively. The GPs most frequently referred to a physiotherapist (51%). CONCLUSION: Once neck pain has become chronic, the minority (44%) of patients do seek help from their GP on a yearly base. In spite of the fact that the patients' conditions are non-specific and chronic, GPs still find indications for further diagnostics in two-thirds of patients. The GPs were rather consistent in their management, as the nature of the diagnostic/therapeutic modalities and referrals was similar in more than 50% of the patients. PMID- 10674299 TI - Allergological examination in general practice and in the specialist field of allergology. A comparative study of the concordance between a group of general practitioners and the allergological specialised health care services to which they refer. AB - OBJECTIVE: To study the concordance between specially trained general practitioners (GPs) and specialist doctors working as consultants to GPs, with regard to diagnosis of allergic illnesses, evaluation of indications for hyposensitisation, and referrals from GPs to specialists. DESIGN: Thirty-four GPs and five specialists practising privately and eight allergologic outpatient clinics participated. The patients included had symptoms indicating allergologic examination. An identical diagnostic procedure was used by the GPs and by the specialists/outpatient clinics. SETTING: General practices, specialist practices, outpatient clinics. SUBJECTS: One-hundred and forty-eight patients. MAIN OUTCOME MEASURES: With regard to skin-prick test there was concordance between the GPs and the specialists in 82.5% of 1322 paired comparisons, in 74.0% with regard to anamnesis + skin-prick test, and in 66.5% with regard to the statement allergy. There was concordance with regard to indication for hyposensitisation in 88.3%, and for referral in 54.1%. There was symmetry concerning prick test and anamnesis + prick test, and asymmetry concerning the statement allergy, and indication for hyposensitisation and for referral. CONCLUSIONS: Specially trained GPs diagnosed specific allergy in concordance with specialists. There was asymmetry between GPs and specialists concerning the statement allergy, indication for hyposensitisation and for referral. PMID- 10674300 TI - Ways of experiencing asthma management. Variations among general practitioners in Sweden. AB - OBJECTIVE: To identify and describe variations in ways of experiencing asthma management among general practitioners (GPs) in Sweden. DESIGN: Descriptive and explorative study using a phenomenographic approach. Semi-structured tape recorded face-to-face interviews, focusing on the GP's own experiences regarding asthma management. SETTING: Primary health care. SUBJECTS: Twenty GPs (12 men, eight women) from 16 health centres in seven counties in central Sweden, 15 from rural areas and small cities and five from medium sized cities or Stockholm suburbs. MAIN OUTCOME MEASURES: Descriptions of ways of experiencing asthma management. RESULTS: Four categories were identified, which described qualitatively different ways of experiencing asthma management: A. The GP focuses on transferring factual knowledge to the patient; B. The GP primarily addresses the patients' application of knowledge in management of the disease; C. The GP aims at improving the patients' understanding of the disease and its management as a prerequisite for self-management; D. The GP concentrates on how to maintain/improve the patient's quality of life despite the asthma disease. CONCLUSION: The GPs describe their ways of experiencing asthma management in qualitatively different ways, likely to have implications for patient care and educational programmes on asthma for GPs. PMID- 10674301 TI - Detection of chronic obstructive pulmonary disease (COPD) in primary health care: role of spirometry and respiratory symptoms. AB - OBJECTIVE: To evaluate the role of spirometry and respiratory symptoms in the detection of chronic obstructive pulmonary disease (COPD) in primary health care. DESIGN: A cross-sectional study. SETTING: A primary health centre in Landskrona, southern Sweden. SUBJECTS: 164 subjects who in 1992 had answered a postal questionnaire concerning obstructive pulmonary diseases and respiratory symptoms. They were aged 45-64 years, with a mean of 55 years. MAIN OUTCOME MEASURES: In 1997, the subjects were invited to perform a spirometry and a medical examination and to answer the same questionnaire as in 1992. Subjects with a forced expiratory volume in 1 second (FEV1) < 85% of the predicted normal value performed reversibility tests. RESULTS: 131 subjects participated in the examinations. 15 subjects (11.5%) were diagnosed as having COPD. Only three of them had been previously diagnosed as having a respiratory disease. Many commonly occurring respiratory symptoms were associated with a reduction in FEV1. CONCLUSIONS: Spirometry examinations in primary health care improve the probability of detecting COPD. A spirometry examination should be considered for patients with respiratory symptoms. It should also be considered for middle-aged smokers, even if they are symptom-free. PMID- 10674302 TI - Effects of an education programme to change clinical laboratory testing habits in primary care. AB - OBJECTIVE: The aim of this study was to investigate the use of clinical laboratory tests in primary care and to evaluate if it is possible to improve the cost-effectiveness of laboratory testing by a short-term education programme. Our main goal has been to lower the total costs of care. DESIGN: An education programme that was monitored by laboratory test ratios. SETTING: Primary health care. SUBJECTS: 63 primary care doctors at 19 primary care centres in the county of Uppsala, Sweden. MAIN OUTCOME MEASURES: The effects of the education programme were monitored by laboratory test ratios (e.g. ASAT/ALAT) of individual doctors before and after the education programme. RESULTS: The education programme resulted in significant changes for the majority of the ratios studied. The savings on direct laboratory costs were approximately SEK 400,000 for assays that were recommended to decrease. The increased cost for assays that were recommended to increase was approximately SEK 140,000 but was considered cost-effective. CONCLUSION: It is possible to achieve significant changes in clinical chemistry test ordering habits of primary care doctors with a 2 day education programme. It resulted in cost savings and a better use of clinical chemistry tests in primary care. The effects were sustained for at least 6 months. PMID- 10674303 TI - Insulin treatment for poorly regulated diabetic patients in general practice. Better regulation and symptom relief? AB - OBJECTIVE: To study the relationship between symptoms, metabolic control and insulin treatment in diabetes mellitus. DESIGN: A cross-sectional questionnaire study of diabetic patients, and a 1-year follow-up study of poorly regulated patients prescribed insulin. Regulation criteria were predefined and the patients acted as their own controls. SETTING: Five primary care practices in Nordland county, Norway. PATIENTS: 111 patients in the cross-sectional study, with 18 of them participating in the follow-up study. MAIN OUTCOME MEASURES: Symptom scores and sum scores, based on five general symptoms--dizziness, depression, fatigue, thirst and dry mouth--and on two urinary symptoms--urinary frequency and urination during the night. RESULTS: Poorly regulated diabetic patients had a higher sum score for general symptoms than better regulated patients (6.1 vs. 4.2, p = 0.078 with Wilcoxon two-sample rank sum test). With parametric analysis, the difference was significant, and remained so when adjusted for age and sex. Females reported more symptoms than males. Symptom relief with insulin was not statistically significant; however, there was a slight but consistent tendency towards less symptoms with better regulation. There was no correlation with HbA1 values at any time during the follow-up study. Only one patient wanted to stop taking insulin after 1 year. CONCLUSION: Better metabolic control and better general well-being seem to be achieved in many cases when poorly regulated patients with type 2 diabetes mellitus begin insulin treatment. The general practitioner should be cautious in promising relief of specific symptoms. PMID- 10674304 TI - As a GP--will I learn anything? PMID- 10674305 TI - [Molecular medicine reaches adulthood in 1999: from the search for the needle in the haystack to genome navigation]. PMID- 10674306 TI - [Shark cartilage on the Internet]. PMID- 10674307 TI - [Prospects for the aged in new millennium]. PMID- 10674308 TI - [New developments in child protection--towards distinct rewards]. PMID- 10674309 TI - [On new antiviral drugs, new viruses and old known bacteria]. PMID- 10674310 TI - [HIV 1999: suppression at what price?]. PMID- 10674311 TI - [Neurology: towards the end of the millenium the end of the decade of the brain]. PMID- 10674312 TI - [Progress in the understanding of the pathogenesis and prevention of bacterial otitis media and meningitis]. PMID- 10674313 TI - [Skin under pressure: irradiated, wounded, infiltrated]. PMID- 10674314 TI - [Refractive surgery in vision disorders: new development and proofs]. PMID- 10674315 TI - [Persistent atelectasis]. PMID- 10674316 TI - The importance of interpreters to insure quality of care for migrants. PMID- 10674317 TI - Addressing language barriers to health care, a survey of medical services in Switzerland. AB - Two descriptive, quantitative cross-sectional surveys including all services of internal medicine and psychiatric services examined how Swiss medical services address the problem of language barriers in health care and how they respond to the high number of allophone patients. Of all the medical services (MS), 244 responded to the questionnaire (Internal medicine: 166; Psychiatry: 78; overall response rate 86.6%). Half of them (51%) estimated the proportion of allophone to the total number of patients at 1-5%. Only 4% of the MS collected statistics on the number of allophone patients (2 internal medicine, 8 psychiatric services). A third of the MS perceive communication with allophone patients as significantly difficult. Only 14% often use qualified interpreters, while 79% often use relatives, 75% often health staff, 43% often employees. Qualified interpreters are less frequently used in internal medicine than in psychiatry. There is an expressed need for qualified interpreters speaking Albanian, Bosnian/Serbo-croat, Tamil and Kurdish. Only 11% of the studied MS have a budget for interpreters, and 17% have access on an interpreter service. 48% express the need to have access to interpreter services. There is a need to raise the awareness of health professionals on the advantages of having access to trained interpreters and on the limits of using relatives as translators. This calls for coordination at national level, policy development and training, in order to ensure adequate communication and quality care for migrants. PMID- 10674318 TI - [When patients and doctors don't speak the same language-- concepts of interpretation practice]. AB - In the Swiss Health Care System, working with an interpreter while medically treating migrants speaking a foreign language has become a common practice. However, the theoretical and conceptional framework of this practice has not yet been sufficiently elaborated. Specifically, the presence of an interpreter leads to a triadic interviewing situation that has implications for the medical treatment. Though an interpreter can improve and deepen the understanding between patient and physician, the communication can also become more complex and difficult with his or her presence. The article sheds light on the perspectives that arise from this situation by defining different role models for medical interpreters and discussing their advantages and disadvantages. The discussion focuses on the four following ideal types: word-for-word translation, cultural mediation, patient advocacy and co-therapy. The choice of interpreter's role in influenced by the institutional setting, the kind of treatment chosen as well as by the concrete interaction. To illustrate the different roles as they manifest in the therapeutic situation, examples from taped and retranslated interviews are presented. Doctor-patient interviews will illustrate the different roles interpreters take over while interpreting medical interviews with migrant patients speaking a foreign language. PMID- 10674319 TI - [Migrant populations at the Hopital de l'Enfance of Lausanne (HEL): evaluation inquiry, taking care and contribution of cultural mediators-translators]. AB - The number of patients belonging to migrant populations who consult Lausanne's Hopital de l'Enfance (HEL) has increased massively in an exponential manner. HEL is a facility dedicated to children with a vocation of public health and training in a university setting and it has to make a point of developing an adequate and pertinent health care system for these populations. In order to do this, a study was undertaken in the form of a prospective survey including over a thousand ambulatory patients. Administrative data (origin, date of migration, nature of the permit and legal situation), social data (home language, professional situation, number of siblings), medical (diagnosis) and psychosomatic data (sleep, enuresis) was recorded as well as data testing the level of understanding between carrier and patient. The study allowed us to define priorities of intervention: introduction of a service of translators--cultural mediators trained by the association "Appartenances" (as well as on going follow-up and assessment of this service), training of health care workers in the fields of cultural mediation and the different aspects of migrant population medicine and, finally, the creation of a steering group within the Institution. Having allowed rapid and spectacular improvement in the quality of the service provided by the HEL, this process is also included in the will to improve health care given to migrant populations at local and national levels in accordance with the priorities defined by the WHO in this domain. It is this experience which is reported in this paper. PMID- 10674320 TI - [Health care networks, migration and cross-cultural adaptation in Lausanne: an action-research in progress]. AB - In Lausanne, Switzerland, there is a growing population of migrant people of different origins. This evolving situation calls for a continuous adjustment between need and offer in terms of healthcare. Up to now, this adjustment, which involves cross-cultural adaptation processes, was based on the use of untrained interpreters. However, clinical experience shows that the use of untrained interpreters tends to keep migrant patients in an unfavourable position. This paper describes an action-research in Lausanne, which aims at the evaluation of the changes that are brought by the introduction of trained cultural mediators and interpreters (CMI) into the medical field. The paper enumerates the clinical issues that gave birth to the project and the methodological choices that were made. After discussing the first results, the authors describe how the different research stages are adapted and modified through continuous mutual influences between the field and the research process itself. PMID- 10674321 TI - Medical interpreters have feelings too. AB - All 22 members of the interpreter service of the Geneva Red Cross were invited to answer an anonymous questionnaire with questions about their work with refugees and asylum seekers. Five (28%) reported having been exposed to a major traumatic event such as war, torture, detention, being beaten. Seven interpreters reported that more than 50% of their sessions involved patients exposed to violence. Five interpreters (28%) frequently experienced difficult feelings during sessions. Twelve (66%) had frequently painful memories. The proportion of interpreters having painful feelings and symptoms increases with the number of sessions with victims of violence. Interpreters also expressed a strong need to talk and share feelings after the session with the medical doctor (83%) or with relatives or spouse (44%). Fifteen (83%) reported seeing patients again outside the consultation. Doctors should be aware of these pressures and give time to interpreters to share their feelings and emotions, to help them cope with their reactions. PMID- 10674322 TI - Language difficulties in an outpatient clinic in Switzerland. AB - This small-scale study attempts to examine the languages spoken in medical consultations during a one-month period in an outpatient clinic in Geneva and the ways health professionals use to communicate with their allophone patients, in particular by using interpreters. Patients of foreign origin accounted for 58% of all the consultations during the survey. Of these, 37% were Non-French-speakers (NFS). The four major language groups of NFS were Albanian, Somali, Tamil and Serbo-croat. Qualified interpreters were used in 24% of the consultations, relatives acting as interpreters in 17%, and in the other consultations without anyone interpreting (59%), a common language had to be negotiated: French, English, Italian, Spanish or German. In only 14% of the consultations without interpreters, both patient's and doctors ability to speak a common language was rated as good. Our data suggest that there has been an increasing awareness of the possible language barriers in the medical outpatient clinic. Even if proxy solutions (informal interpreters or the use of a common language) still play an important role, access to an interpreter service has been widely used. This calls for systematic and regular interpreter use, planning the interpreting needs in a timely manner. In the future, training in working with interpreters should become an integral part to the introductory sessions for the junior physicians assigned to the outpatient clinic. PMID- 10674323 TI - [Cultural mediation and training of health care professionals: from cross-culture to co-discipline]. AB - Why organize a training course destined for health care professionals which is specifically devoted to cultural mediation in the sphere of health care? There are several reasons justifying such an initiative, mainly relating to the social and cultural mutations pertaining to the present world situation. In fact, two major phenomena may be said to directly influence the health care scene in the current context of generalized migration: the internationalization of diseases on the one hand and on the other hand, the cultural plurality to be found to an increasing degree in society in general. While the responses to the first scenario fall within the limits of standard medical practice, those which are necessitated by the second situation demand a widening of expertise. By reason of the relationship they establish with migrants, it is mandatory that carers should modify their own personal attitudes as well as their professional framework. In the face of the requirements imposed by cross-culture, it is imperative to move in the direction of co-discipline. PMID- 10674324 TI - Explanations of unmet need for contraception in Chitwan, Nepal. AB - This article explores reasons why women's fertility preferences and their contraceptive behaviors often appear to be contradictory. Ninety-eight separate interviews with women and their husbands conducted in rural Chitwan District, Nepal, over a 12-month period in 1993-94 revealed that people continually and self-consciously weigh the perceived benefits and risks of practicing family planning relative to their situations. Temporary and, especially, hormonal methods are perceived to carry unacceptable health risks. Contraceptive technologies are evaluated in relation to competing priorities and interests. Household poverty heightened the perceived risk of family planning use; poor people fear they can ill afford negative effects to their health that might result. People assess their health status and physical workload, factors that they believe condition their ability to use family planning methods without experiencing damaging health effects. Strategies employed to lower contraceptive risk include altering the method of choice, manipulating relationships with spouses, timing the adoption of contraceptives, managing the context of service provision, and acting in light of the experiences of others. Qualitative findings from the fieldwork are complemented by analysis of data from a standardized fertility survey. PMID- 10674325 TI - Understanding the prevalence of female sterilization in rural south India. AB - By analyzing the practice of female sterilization in rural Andhra Pradesh, in southern India, this article examines the role culture plays in demographic research. The popularity of female sterilization in rural Andhra Pradesh is shown to be intelligible if the symbolic value of a young mother's reproductive capacity is understood in terms of familial power relations. Through sterilization, young mothers can symbolically push their influential mothers-in law toward old age, thus increasing their own relative prestige, and they can strive to control the ambiguity surrounding their reproductive functions. This study is based on 14 months of participant observation in three rural villages, a survey of 396 households, and unstructured interviews with 42 women and two men. It shows how demography and anthropology can be mutually supportive in their efforts to clarify population phenomena. PMID- 10674326 TI - Household organization, women's autonomy, and contraceptive behavior in southern Ethiopia. AB - The Southern Nations, Nationalities, and People's Region of Ethiopia (SNNPR) is home to 11 million people constituting more than 45 language and ethnic groups, most of whom live in extremely poor rural communities. Data for currently married, fecund women aged 15-49 from demographic surveys conducted in the SNNPR in 1990 and 1997 are used to investigate contraceptive knowledge and communication, and the use and future need for family planning services in this population. This study focuses on how these processes are affected by household organization and women's status, and on their implications for population policies and programs. Considerations of the implications of these results for understanding the fertility transition of a highly diverse African population under severe stress are presented. Although household extension and polygamy characterize one-third of the women sampled, they do not affect the women's contraceptive behavior. Women's literacy and autonomy are, by far, the most significant forces in the movement toward lower fertility in the region. PMID- 10674327 TI - Contraceptive switching in Bangladesh. AB - Bangladesh has experienced a substantial decline in fertility that has been achieved by means of a large increase in the use of modern methods of contraception. As contraceptive prevalence increases, aspects of contraceptive use dynamics, including reasons for discontinuation and behavior after discontinuation, become important influences on fertility. This report uses calendar data from the 1993-94 Bangladesh Demographic and Health Survey to examine contraceptive behavior following discontinuation of modern-method use. The individual-level characteristics found to influence switching behavior include the method used, method-related difficulties with previous contraceptive use, and education. A large amount of unexplained variation in switching rates remains, however, largely at the individual level, but also at the community level for certain types of transition. PMID- 10674328 TI - Maternal mortality in Vietnam in 1994-95. AB - This report presents the first population-based estimates of maternal mortality in Vietnam. All the deaths of women aged 15-49 in 1994-95 in three provinces of Vietnam were identified and classified by cause. Maternal mortality was the fifth most frequent cause of death. The maternal mortality ratio was 155 deaths per 100,000 live births. This ratio compares with the World Health Organization's estimates of 430 such deaths globally and 390 for Asia. The maternal mortality ratio in the delta regions of these provinces was half that of the mountainous and semimountainous regions. Because a larger proportion of the Vietnamese population live in delta regions than elsewhere, the maternal mortality ratio for Vietnam as a whole may be lower than that of the three provinces studied. Maternal mortality is low in Vietnam primarily because a relatively high proportion of deliveries take place in clinics and hospitals, where few women die in childbirth. Also, few women die of the consequences of induced abortion in Vietnam because the procedure is legal and easily available. PMID- 10674329 TI - Assessing recall and understanding of informed consent in a contraceptive clinical trial. PMID- 10674330 TI - Kyrgyz Republic 1997: results from the Demographic and Health Survey. PMID- 10674331 TI - Yemen 1997: results from the Demographic and Health Survey. PMID- 10674332 TI - [Risk factors for posttraumatic fits and epilepsy]. AB - There is still a considerable controversy about the usefulness of antiepileptic prophylaxis after traumatic brain injury. Overall incidence of posttraumatic fits and epilepsy's is well known, but an individual decision on prophylaxis requires knowledge about the individual risk. We performed a prospective observational study on 612 patients with traumatic brain injury of every degree of severity. Follow-up by phone call included 96.2% of the study population after 6 month and 91.2% after 36 month, respectively. The overall incidence for early fits (within 7 days after trauma), late fits (up to 36 month) and epilepsy (as defined by the International League Against Epilepsy) was 4.2%, 3.7% and 2.5%, respectively. These incidences increased according to the severity of the trauma, but the most powerful single predictor was intracranial hemorrhage. There was no significant difference related to the hemorrhage localisation. Development of epilepsy was much more common after late fits (48%) than after early fits (17%). These features established a hierarchy of risks for the development of epilepsy: no intracranial hemorrhage/no fit: 1% (4/437), intracranial hemorrhage/no fit: 8% (10/122), intracranial hemorrhage/early fit: 16% (3/19), intracranial hemorrhage/late fit: 53% (7/13). If prophylactic antiepileptic treatment is desired, but should be restricted to patients at high risk. These are patients with intracranial hemorrhage--are a well defined high risk group--when their first fit is a late fit. PMID- 10674333 TI - [Neuropsychological fields in early neurotrauma rehabilitation]. AB - It meanwhile is commonly accepted that early onset of specific rehabilitation intervention in traumatic brain injured patients will enhance the recovery of brain function. The integration of neuropsychology in the early treatment of traumatic brain injury can mainly be ascribed to the increasing recognition that cognitive, personality and emotional deficits have been the most devasting longterm problems faced by patients and their families. The aim of our paper is to illustrate the role of neuropsychology in the early stage of rehabilitation. Neuropsychological therapy and the application of appropriate tests depend on the level of consciousness and the extent of behavioural problems. Observation, cognitive screening tests and the use of valid neuropsychological tests make up the main approaches. Our rehabilitation program includes measures of sensory and cognitive stimulation. Improvement of attention and stimulation of cognitive functions are one of the most important aims of early neuropsychological therapy. We choose tasks which require automatic information processing, the retrieval of well established knowledge and implicit learning. Appropriate tests and the development of neuropsychological treatment programmes represent an important means of maximising the patient's capacity to benefit from early rehabilitation. PMID- 10674334 TI - Management of vein of Galen malformations. A review based on five neurosurgically treated cases and literature reports. AB - The high morbidity and mortality associated with the management of vein of Galen aneurysmal malformations (VGAM) continues to pose a tremendous challenge to the neurosurgeon as well as to the attending interventional radiologist. Since 1985, five patients with VGAM have been referred to the neurosurgical unit of the University of Cologne, two neonates, one infant and two adults. Four patients underwent direct operation and two patients received a shunt. The treatment was performed without mortality. A review of the literature reflects no substantial difference between neurosurgical treatment during the last 15 years (mortality 10%) and endovascular treatment (best series mortality 6%). PMID- 10674335 TI - [Value of German Hospital Diagnosis Statistics in epidemiology-- an analysis with subarachnoid hemorrhage as an example]. AB - Due to the Hospital Statistics Regulations of the 10th April 1990, the German Hospital Diagnosis Statistics were introduced in 1993 with the intention to serve as a database of health care decisions. This purpose requires a high-quality collection of epidemiologically relevant data. From 1993 to 1996 we analysed the datasets obtainable for subarachnoid haemorrhage which is coded with ICD 430. A subset concerning the data of 1996 and the Saarland region was compared to the data of the Saarland medical school at Homburg/Saar. Cases treated in the neurosurgical department were critically reviewed. About 20% of the cases coded with ICD 430 showed no subarachnoid haemorrhage. On the other hand, again about 20% of subarachnoid haemorrhage cases were not coded with ICD 430. The statistics comprise duplicates due to transfers between hospitals. The calculation of incidence is not possible because new bleeding cases cannot be outlined. In the present form the German Hospital Diagnosis Statistics are not suitable as a reliable base of health care decisions. This is partly caused by the inadequacy of the ICD-classification but, also, by the criteria for collecting data. We propose several modifications which can improve data quality in order to meet the intended requirements. PMID- 10674336 TI - [Paralysis of the foot as the first symptom of a herniated thoracic disc]. AB - Herniated thoracic discs are uncommon entities that are difficult to diagnose. They may be associated with a myriad of symptoms, which often delays diagnosis. In general dorsal pain that radiats around the chest or abdomen are found. In case of spinal cord compression signs of thoracal myelopathy are common. This paper describes a patient with a complete paralysis of the left foot as the first symptom of a herniated thoracic disc. After frustrated diagnostic of the lumbar spine, the exact neurological examination showed a sensible cut at the level of D6/7, the MR tomography diagnosed a herniated thoracic disc at the same level. Four months later he was presented again with a plegia of the left foot and a sensible cut at the level D5/6. The MR tomography showed a herniated disc at the level above the spondylodesis. The immediately performed transthoracic disc excision and fusion of D6/7 was followed by complete remission of the plegia and the sensible cut. Four months later we performed a rethoracotomy, disc excision and decompression with a spondylodesis D5/6. The procedure was again followed by complete remission. In case of paralysis of the lower extremities one has to consider a herniated thoracic disc. PMID- 10674337 TI - Evaluation of abstracts submitted for the annual meeting of the German Neurosurgical Society 1999--unravelling a mystery. AB - The evaluation for the abstracts submitted for the annual meeting of the German Neurosurgical Society together with the Swiss Neurosurgical Society in Munich 1999, is presented as it has developed during the meetings of the last years. 597 abstracts were reviewed by the 30 members of the review committee according to a 5 point grading system. Cut off for acceptance was a mean grading of 2.7 points. Abstracts better than 2.4 were accepted, abstracts worse than 2.7 were rejected. Experimental studies were judged slightly better than clinical studies: the mean grading of clinical and experimental studies was 2.55 +/- 0.5 vs. 2.72 +/- 0.6 (p < 0.02). All abstracts with a mean grading of 2.4-2.7 and a standard deviation > 1.0 were discussed in a meeting of the review committee. 353 abstracts were accepted. Some of the abstracts submitted for oral presentation had to be converted to poster presentations. Among others the decision was based on the grading of the abstract. PMID- 10674338 TI - [Good practice: guidelines for neurosurgery. European Association of Neurosurgical Societies World Federation of Neurosurgical Societies]. PMID- 10674339 TI - CMV in HIV-infected newborns. PMID- 10674340 TI - Telomerase, immortality, and cancer. PMID- 10674341 TI - The adenovirus and bronchopulmonary dysplasia: an association that could be causal or coincidental. PMID- 10674342 TI - Thrombopoietin and neonatal thrombocytopenia. PMID- 10674343 TI - Neural crest-derived defects in experimental esophageal atresia. AB - Esophageal atresia (EA) is often associated with cardiovascular and other malformations that are likely neural crest derived. The present study tests the hypothesis that the heart and great vessels and the thymus and parathyroids may be abnormal in the rat model of EA as a result of disturbed neural crest development. Time-mated pregnant rats received intraperitoneally on d 8 and 9 of gestation either 2 mg/kg adriamycin or vehicle. Esophageal, heart, and thymic malformations were sought under the microscope in term fetuses. The parathyroids were histologically investigated. Control fetuses had no malformations, whereas 69 of 109 fetuses exposed to adriamycin had EA and 45 of 69 had 15 right aortic arches, nine aberrant right subclavia, eight ventricular septal defects, six narrow pulmonary outflow tracts, five tetralogies of Fallot, three double outflow right ventricles, three double aortic arches, three atrial septal defects, three right ductus arteriosus, and two truncus. The thymus was absent in 19, hypoplastic in 12, and ectopic in five out of 36 fetuses with EA in which it was studied, whereas the parathyroid glands were absent in 16, single in four, and ectopic in one of the 23 fetuses with EA in which they were studied. In conclusion, the nature of the cardiovascular, thymic, and parathyroid malformations associated with EA in rats is consistent with the hypothesis of neural crest participation in their pathogenesis. Mechanisms simultaneously disturbing foregut septation, somitic segmentation, and neural crest development should be sought to explain the combined occurrence of malformations in EA. PMID- 10674344 TI - Discoordinate expression of pancreatic lipase and two related proteins in the human fetal pancreas. AB - The lipase gene family contains a large number of members. Among the most closely related are pancreatic triglyceride lipase (PTL) and two pancreatic lipase related proteins (PLRP1 and PLRP2). Previous studies in rodents demonstrated divergent temporal expression of the genes encoding these proteins. PLRP1 and PLRP2 were expressed in fetal pancreas, whereas PTL was not expressed until pups were several weeks old. To determine whether the human pancreas has a similar expression pattern for these genes, we determined the levels of each mRNA in fetal pancreas at various ages. A reverse transcriptase-PCR method was developed and used to quantify the mRNA levels for the three species normalized to the mRNA encoding cyclophillin. The mRNA encoding PLRP1 and PLRP2 was present by 16 wk in the fetal pancreas. In contrast, the mRNA encoding PTL was not present in the fetal pancreas. This pattern of expression suggests that the genes encoding theses proteins have different regulatory elements controlling temporal expression and provides another example of nonparallel expression of genes encoding pancreatic exocrine proteins. PMID- 10674345 TI - Regulation of fetal rat bone growth by C-type natriuretic peptide and cGMP. AB - C-type natriuretic peptide (CNP) and its high affinity receptor-B are expressed in fetal bones. Here we show that CNP accelerates longitudinal growth of fetal rat metatarsal bones in organ culture by several mechanisms. First, CNP stimulates chondrocyte proliferation in the proliferative zone as assessed by [3H]thymidine incorporation. Second, CNP stimulates cell hypertrophy as assessed by quantitative histology. Third, CNP stimulates cartilage matrix production as assessed by incorporation of 35SO4 into glycosaminoglycans. Natriuretic peptide receptor-B contains an intracellular guanylyl cyclase catalytic domain. We therefore hypothesized that cyclic GMP (cGMP) would reproduce the effects of CNP on fetal bones. Consistent with this hypothesis, we found that 8-Br-cGMP, like CNP, stimulates longitudinal growth and glycosaminoglycan synthesis. However, unlike CNP, cGMP inhibits proliferation of growth plate chondrocytes and has no effect on hypertrophy. We conclude that CNP stimulates longitudinal bone growth by increasing chondrocyte proliferation, chondrocyte hypertrophy, and cartilage matrix production. cGMP, a second messenger for CNP, reproduces some but not all of the effects of CNP, suggesting that other signal transduction mechanisms may also be involved. PMID- 10674346 TI - Milk-borne epidermal growth factor modulates intestinal transforming growth factor-alpha levels in neonatal rats. AB - Epidermal growth factor (EGF) is present in milk from various mammalian species, but its physiologic function in neonatal development remains unclear. Transforming growth factor-alpha (TGF-alpha) is a peptide structurally related to EGF, and its presence is detected in the developing small intestine of rats. The purpose of the present study was to examine the effect of milk-borne EGF on endogenous production of EGF and TGF-alpha in the small intestine of suckling rats. Neonatal rats were fed via gastrostomy either growth factor-free rat milk substitute (RMS) or RMS supplemented with EGF (100 ng/mL of RMS) from 8 to 12 d of age. Artificially reared rats were then compared with their dam-fed littermates. Animals fed the EGF-deficient diet RMS had markedly increased EGF and TGF-alpha mRNA levels in duodenum and ileum compared with dam-fed controls and significantly elevated total intestinal content of TGF-alpha peptide. Intestinal EGF content and EGF serum levels were significantly decreased in the RMS group compared with controls. The addition of EGF to the RMS diet normalized TGF-alpha mRNA levels in the duodenum and ileum, EGF mRNA levels in the ileum, and total intestinal TGF-alpha content and EGF serum levels to the levels measured in dam-fed littermates. Motility studies showed that enteral administration of EGF did not affect stomach emptying and intestinal transit. These studies indicate that exogenous milk-borne EGF modulates endogenous production of TGF-alpha in developing small intestine. It is likely that neither TGF-alpha nor EGF are solely responsible for small intestinal overgrowth of artificially reared neonatal rats. PMID- 10674347 TI - Increased local synthesis of epidermal growth factors in infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis (IHPS) is characterized by hypertrophy of the pyloric muscle. The growth of smooth muscle cells is regulated by several growth factors. Epidermal growth factor (EGF) and heparin-binding EGF-like growth factor are potent mitogens for smooth muscle cells. In the present study, we investigated immunohistochemical localization of EGF and EGF-related peptides and EGF mRNA expression in pyloric smooth muscle cells to determine whether the EGF family is involved in the process of pyloric muscle hypertrophy in IHPS. Pyloric muscle biopsy specimens were obtained at the time of pyloromyotomy from 10 patients with IHPS. Control material included 10 pyloric muscle specimens taken at autopsy from age-matched cases without evidence of gastrointestinal disease. Indirect immunohistochemistry was performed using the avidin-biotin-peroxidase complex method with anti-EGF, anti-EGF receptor, and anti-heparin-binding EGF like growth factor antibody. In situ hybridization was performed using digoxigenin-labeled EGF-specific oligonucleotide probe. The pattern of immunoreactivity in pyloric muscle with EGF, EGF receptor, and heparin-binding EGF-like growth factor was similar in all specimens. There was a marked increase in EGF, EGF receptor, and heparin-binding EGF-like growth factor immunoreactivity and EGF mRNA expression in smooth muscle cells in pyloric circular and longitudinal muscle from patients with IHPS compared with control specimens. These data suggest that the upregulated local synthesis of EGF and EGF-related peptides in pyloric muscle may play a critical role in the development of pyloric muscle hypertrophy in IHPS. PMID- 10674348 TI - Pharmacokinetics, pharmacodynamics, and safety of administering pegylated recombinant megakaryocyte growth and development factor to newborn rhesus monkeys. AB - Thrombocytopenia is common among sick neonates. Certain groups of thrombocytopenic adults respond favorably to the administration of recombinant thrombopoietin or to pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF), a recombinant human polypeptide that contains the receptor-binding N-terminal domain of thrombopoietin. The effectiveness and safety of such treatment in neonates, however, have not been reported. The purpose of the present study was to determine the biologic activity and safety of PEG-rHuMGDF administration to newborn rhesus monkeys. Eight monkeys were divided into four groups and treated subcutaneously with 0.00, 0.25, 1.00, or 2.50 microg/kg once daily for 7 d. Complete blood counts, serum chemistries, clotting panels, and MGDF levels were followed serially, and hematopoietic progenitor cell assays were performed on bone marrow aspirates before the first dose and again on d 8. Pharmacokinetic evaluations were performed on the animals that received the highest dose of PEG-rHuMGDF. All monkeys had normal growth during the study period, and all chemistries, clotting studies, and blood pressure measurements were normal. The peak serum MGDF concentration occurred at 3 h, and the half-life was 8.4 to 13.0 h. As in adult rhesus monkeys, platelet counts in the treated neonates began to rise on d 6, peaked on d 11, and returned to baseline by d 23. The two highest doses generated an 8- to 12-fold increase in platelets, whereas those treated with 0.25 microg/kg had a 6-fold increase. Other hematologic parameters measured were unaffected. Thus, newborn monkeys responded to doses of PEG-rHuMGDF that were similar to or smaller than (per kilogram body weight) those that are effective in adult animals and did so without obvious short-term toxicity. PMID- 10674349 TI - Influence of maternal smoking on autonomic nervous system in healthy infants. AB - To determine the influence of maternal smoking on autonomic nervous system in healthy infants, 36 infants were recorded polygraphically for one night. Their mothers were defined, according to their smoking frequency during pregnancy, as "nonsmokers" (no cigarettes smoked during pregnancy) or "smokers" (10 or more cigarettes per day). The infants had a median postnatal age of 10.5 wk (range 6 to 16 wk); 18 were born to nonsmokers, and 18 to smokers. During the whole night, spectral analyses of heart rate (HR) were evaluated as a function of sleep stages. Two major peaks were recognizable: a low-frequency component (LF) related to sympathetic and parasympathetic activities and a high-frequency component (HF) reflecting parasympathetic tonus. The ratio of LF/HF powers was calculated as an index of sympathovagal interaction. In REM sleep, "smokers" infants were characterized by significantly lower HF powers and normalized HF powers, and higher LF/HF ratios than "nonsmokers." The finding did not reach statistical significance in NREM sleep. In conclusion, maternal smoking induced changes in autonomic control and maturation in infants. These effects of cigarette smoke exposure can be added to those already reported and offer additional evidence for counseling mothers to stop smoking. PMID- 10674350 TI - Total hydroperoxide and advanced oxidation protein products in preterm hypoxic babies. AB - Previous studies have shown that plasma lipoproteins are a common target of free radical-induced oxidative stress in hypoxic newborn infants. In contrast to lipids, the reaction of proteins with various oxidants during hypoxia has not been extensively studied. We tested the hypothesis that tissue hypoxia results in increased production of protein oxidation in cord blood of preterm newborns. Heparinized blood samples of 39 hypoxic and 16 control preterm newborns were obtained from the umbilical vein, after cord clamping immediately after delivery. Plasma levels of total hydroperoxide (TH), advanced oxidation protein products (AOPP), hypoxanthine (Hx), xanthine (Xa), and uric acid (UA) were measured. Higher Hx, Xa, UA, TH, and AOPP levels were found in hypoxic newborn infants than in controls. Statistically significant correlations were observed between: TH and Hx (r = 0.54, p = 0.003, n = 28), AOPP and Hx (r = 0.64, p = 0.0001, n = 27), and TH and AOPP plasma levels (r = 0.50, p = 0.02, n = 21). In summary, TH, AOPP, Hx, Xa, and UA production is increased in fetal blood during hypoxia. The more severe the hypoxia, the higher the lipid and protein damage by free radicals. PMID- 10674351 TI - Detection of microorganisms in the tracheal aspirates of preterm infants by polymerase chain reaction: association of adenovirus infection with bronchopulmonary dysplasia. AB - Bronchopulmonary dysplasia (BPD) is recognized as an important cause of morbidity and mortality in preterm infants. Because the role of congenital infections in BPD has been debated, the purpose of this study was to test the hypothesis that detection of infectious agents in tracheal aspirate samples was associated with the development of BPD. Tracheal aspirate samples were obtained within the 1st week of life and screened by polymerase chain reaction for adenovirus, cytomegalovirus, parvovirus, enteroviruses, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma pneumoniae, and Chlamydia species. BPD was defined as persistent oxygen dependence at 28 d of age and 36 wk postconceptional age (PCA). Infants that expired before these time points were excluded from statistical analysis. Out of 89 infants studied, at 28 d of life, 13 had expired, 45 had BPD, and 31 had no BPD (controls). At 36 wk PCA, 15 infants expired, 39 still had BPD, and 35 did not. A significant increase in the frequency of adenovirus genome was identified in BPD patients compared with controls, both at 28 d of life (12/45 = 27% versus 1/31 = 3%: p< or =0.01) and at 36 wk PCA (10/39 = 29% versus 2/35 = 6%: p = 0.01). Other microorganisms were rarely detected and not associated with the development of BPD. This is the first study reporting the frequency of detection of adenovirus DNA in tracheal aspirate samples obtained during the 1st week of life from infants with BPD and suggests that prenatal acquisition may be important in the development of BPD. PMID- 10674352 TI - Measurement of baroreceptor-mediated effects on heart rate variability in fetal sheep. AB - To determine if alterations in arterial pressure influenced fetal heart rate variability (HRV), experiments were carried out in chronically catheterized fetal sheep aged 128-138 d. Arterial pressure was raised or lowered by intravenous infusion of phenylephrine or sodium nitroprusside, and the effects on heart rate (HR) and HRV were measured (HRV, as the coefficient of variation (CV) in mean pulse interval or by power spectral analysis). Experiments were carried out before and during beta-adrenoceptor blockade with propranolol or before and during cardiac vagal blockade with atropine. There were positive relationships between mean arterial pressure and HRV (slope = 0.074+/-0.001, r = 0.81+/-0.06, p<0.001, measured as the CV of pulse interval) and between mean arterial pressure and power spectral density (slope = 4+/-0.5, r = 0.89+/-0.02, p<0.001) in the frequency range 0.04-0.08 Hz. Beta-adrenoceptor blockade had no effect on these relationships, but they were abolished by cardiac vagal blockade. The sigmoid relationship between fetal HR and mean arterial pressure, i.e. the cardiac baroreflex, was affected, however, by blockade of cardiac sympathetics and abolished by blockade of cardiac vagal activity. Thus, fetal HRV was affected by alterations in arterial pressure, and these effects depended on the integrity of the cardiac vagus, not on alterations in cardiac sympathetic activity. Therefore, although baroreflex control of fetal HR depends on the integrity of both sympathetic and parasympathetic efferent pathways, baroreceptor-induced changes in HRV depend only on the cardiac vagus. PMID- 10674353 TI - Lavage administration of dilute recombinant surfactant in acute lung injury in piglets. AB - In an acute lung injury model, we previously observed reversal of pulmonary dysfunction with natural surfactant administered by lavage (dose = 18 mg/kg phospholipid). The present study questioned whether a lower dose of phospholipid would be effective if a recombinant preparation rather than natural surfactant were used. Acute lung injury was induced by repeated saline lung lavage in ventilated, sedated, and paralyzed piglets. Three concentrations of recombinant surfactant were studied (low phospholipid, 1 mg/mL; medium phospholipid, 4 mg/mL; high phospholipid, 13.5 mg/mL). Control piglets received no surfactant. Thirty five milliliters per kilogram of surfactant was administered by gravity, followed by passive drainage of excess fluid. All treatment groups retained similar volumes (4.7+/-0.3 mL/ kg), corresponding to phospholipid doses of 4+/-0.4, 22+/ 3, and 67+/-4 mg/kg in low, medium, and high-dose groups, respectively. Treatment groups showed significant improvement in Pao2 compared with controls. Other parameters different from controls were found in only the medium and high-dose groups. All surfactant-treated groups showed improvement over time in Pao2, Paco2, lung resistance mean airway pressure, functional residual capacity, and dynamic compliance. These data support the statement that whereas there is a dose response to exogenous surfactant, the effective dose of recombinant surfactant in acute lung injury may be as low as 4 mg/kg phospholipid when administered by lavage. PMID- 10674354 TI - IL4 and IL4Ralpha genes are not linked or associated with type 1 diabetes. AB - Previous studies have shown the immunoregulatory functions IL-4 in type 1 diabetes mellitus. Therefore, the genes involved in the IL-4 regulatory pathway are candidates for diabetes susceptibility genes. Here we have evaluated IL4 and the alpha subunit of the IL-4 receptor (IL4Ralpha) genes using the affected sibpair (ASP) and transmission/disequilibrium test (TDT). We analyzed 309 diabetic families from the United States and 87 families from various European countries. There was no evidence that either of these two genes are linked or associated with type 1 diabetes. Means by which IL-4 directed signals could indirectly alter diabetes susceptibility are proposed. PMID- 10674355 TI - Growth hormone action in hypothyroid infant rats. AB - In neonatal rats, expression of serine protease inhibitors 2.1 and 2.3 mRNA peaks on d 2 of life and declines shortly thereafter, coinciding with levels of circulating GH. To evaluate the role of GH in this increase and to test the hypothesis that GH is active in perinatal life, we studied GH action in a model of GH deficiency. Maternal/neonatal hypothyroidism with consequent GH deficiency was induced by methimazole administration to pregnant dams. The resultant hypothyroid neonates were treated at d 2 or 7 of age with GH or saline for 1 h before exsanguination. In d-7 neonates, but not at d 2, GH administration resulted in significant serine protease inhibitors 2.1 and 2.3 mRNA induction. This treatment did not result in increased production of either GH receptor or IGF-I mRNA at either age. There was a slight GH-independent increase in GH receptor and IGF-I mRNA expression by d 7. Electromobility shift assays using hepatic nuclear extracts from these neonates and the GH response element from the serine protease inhibitor 2.1 promoter showed signal transducer and activator of transcription 5 (Stat5) binding in response to GH in extracts from d-7 rats only. Immunoblots of these extracts showed twice as much Stat5 in the nuclei of d-7 treated neonates compared with d-2 treated neonates. We conclude that there is apparent insensitivity to GH treatment in d-2 neonates that remits by d 7 and that this remission correlates with increased abundance of GH receptor and Stat5. PMID- 10674356 TI - Optimization of inspiratory work in periodic breathing in infants. AB - In periodic breathing, there are repeated cycles of bursts of breaths separated by pauses several seconds long. We consider the mechanics of periodic breathing in human infants using calibrated traces of tidal volume and esophageal pressure recorded during the first few days after delivery. Each cycle of periodic breathing was analyzed in terms of the inspiratory time and beginning and end inspiratory volumes for each breath, the number of breaths in the cycle, and the total observed inspiratory work. A simple model was used to characterize the mechanics of the lung during inspiration, and the recordings were used to calculate the parameters of this model. These varied from breath to breath. A theoretical formula is derived for the sum of external work performed during inspiration for each burst. It is shown mathematically that there exists a local minimum in the calculated work as the values of the individual tidal volumes in this formula are allowed to vary, with the constraint that the sum of the ventilation during the cycle is as measured. The measured values of inspiratory timing, the starting volume and pressure, and the mechanical parameters for each inspiration are also used. We show that during each cycle of periodic breathing, the total of the observed external work is highly correlated with this theoretical minimum work. In addition, during the cycle, there is a pattern of overshoot and subsequent undershoot in the work with respect to the theoretical optimum, which suggests a control process operating during the cycle to minimize the work. PMID- 10674357 TI - Expression of gamma-glutamylcysteine synthetase during development. AB - Prematurity has been associated with low glutathione (GSH) concentrations in bronchoalveolar lavage fluid as well as in leukocytes from tracheal aspirates and peripheral blood. To elucidate whether this is caused by deficient GSH synthesis, the expression and activity of gamma-glutamylcysteine synthetase (glutamate cysteine ligase, GCS, EC 6.3.2.2), the rate-limiting enzyme for GSH synthesis, were measured from fetal, neonatal, and adult human liver, lung, and kidney samples. The highest activity was measured in the liver, in which mRNA expression of the catalytic GCS heavy and the regulatory light subunits, as well as activity, were, on average, similar in the various stages of development. Although GCS light subunit mRNA concentrations in the lung were higher in neonates than in fetuses and adults, enzyme activities were similar. In the adult kidney, mean enzyme activity was somewhat higher than in fetal or neonatal kidney, but this may be accounted for by the variation in the small number of samples. In conclusion, GCS is expressed and active already in the second trimester and thus low GSH concentrations found in preterm neonates appear not to be explained by deficient GSH synthesis. Other factors, such as limited availability of the GSH precursor cysteine or increased GSH consumption, may account for the lower concentrations of GSH found in preterm infants. PMID- 10674358 TI - Formation of L-alloisoleucine in vivo: an L-[13C]isoleucine study in man. AB - L-alloisoleucine (2S, 3R), a diastereomer of L-isoleucine (2S, 3S), is a normal constituent of human plasma. Considerable amounts accumulate in maple syrup urine disease, in which the branched-chain 2-oxo acid dehydrogenase step is impaired. The mechanism of L-alloisoleucine formation, however, is unclear. We addressed this issue by performing oral L-[1-13C]isoleucine loading (38 micromol/kg body wt, 50% 1-13C) in overnight-fasted healthy subjects (n = 4) and measuring the 3-h kinetics of 13C-label incorporation into L-isoleucine plasma metabolites. Compared with L-isoleucine, the time course of 13C-enrichment in the related 2 oxo acid, S-3-methyl-2-oxopentanoate, was only slightly delayed. Peak values, amounting to 18+/-4 and 17+/-3 mol percent excess, respectively, were reached within 35 and 45 min, respectively. The kinetics of 13C-enrichment in S- and R-3 methyl-2-oxopentanoate enantiomorphs were similar and linearly correlated (p << 0.001). In L-alloisoleucine, however, 13C-label accumulated only gradually and in minor amounts. Our results indicate that R-3-methyl-2-oxopentanoate is an immediate and inevitable byproduct of L-isoleucine transamination and further suggest that alloisoleucine is primarily formed via retransamination of 3-methyl 2-oxopenanoate in vivo. PMID- 10674360 TI - Neuroimaging of hallucinations: a review of the literature. AB - While hallucinations have been described for over two millennia, their cause remains unclear. Brain-based models suggest that abnormal cerebral excitation and a lack of normal cerebral inhibition may play primary roles, but evaluation of these hypotheses has been hampered by difficulty in studying the hallucinatory state. Recent advances in neuroimaging have provided researchers with tools to study a variety of mental states, including hallucinations. We review the literature regarding the structural and functional neural correlates of hallucinations. Despite small sample sizes and methodological differences, several studies describe similar results: hallucinations are associated with sensory modality-specific activation in cerebral areas involved in normal sensory processing. Furthermore, neural activation may be specifically related to distinct phenomenological features of the hallucinatory experience. Further work is needed to better understand the neural basis of hallucinations. PMID- 10674359 TI - Dietary trans fatty acids affect docosahexaenoic acid concentrations in plasma and liver but not brain of pregnant and fetal rats. AB - The aim of the present study was to investigate the maternal-fetal transport, incorporation, and effects on liver delta-6 fatty-acid desaturase activity of dietary trans fatty acids in pregnant rats. Three groups of six rats each were fed three experimental diets containing approximately 0%, 15%, and 30% of trans fatty acids but containing the same proportion of linoleic (18:2 n-6) and a linolenic (18:3 n-3) acids for 10 wk. On d 20 of pregnancy, the animals from each group were killed. We determined the fatty acid profiles in plasma, brain, and liver microsomes of pregnant rats, as well as in placenta and fetal liver and brain. No changes were found in the number of fetuses of the pregnant rats. Trans fatty acids were incorporated in high concentrations in placenta and in maternal and fetal tissues, except brain, strongly elevating the linoleic acid proportion and lowering that of docosahexaenoic acid. The delta-6 fatty-acid desaturase activity in the liver microsomes of the pregnant rats was inhibited by trans isomers. In conclusion, high intakes of trans fatty acids partially inhibit liver delta-6 fatty-acid desaturase in pregnant rats, which may explain, in part, the low concentrations of docosahexaenoic acid in pregnant and fetal tissues. However, the fatty acid composition of both fetal and pregnant rat brain remains mostly unaffected regardless of the dietary trans fatty acid content. PMID- 10674361 TI - Neo-striatal rCBF correlates of psychomotor slowing in patients with major depression. AB - Psychomotor slowing is a fundamental clinical feature of severe depression and is thought to reflect dysfunction within prefrontal-subcortical circuits. This study utilised a split-dose single photon emission computerised tomography (SPECT) scanning technique in association with a two-stage test of psychomotor speed. Twenty-five patients with primary depressive disorders were injected with technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) whilst performing each component of a two-stage psychomotor task. The first stage, 'simple reaction time' (RT) and the second stage, 'choice reaction time' (CRT), were each followed by 30-min SPECT scans. Regions of interest (ROIs) corresponding to the left and right neo-striatum (caudate-putamen) were drawn, and regional cerebral blood flow (rCBF) values were calculated. Importantly, the change in rCBF measure in the left neo-striatum was inversely correlated with RT (r = -0.48, P < 0.05). That is, the patients with the greatest psychomotor slowing initially showed the least increase in rCBF during the CRT condition. This effect was independent of age. The study demonstrates that a simple two-stage motor paradigm can be used to elicit rCBF correlates of psychomotor slowing in patients with primary depression. Such rCBF findings may implicate the neo-striatum in the neurobiology of major depression. PMID- 10674362 TI - Proton magnetic resonance spectroscopy of lenticular nuclei in obsessive compulsive disorder. AB - Magnetic resonance spectroscopy (MRS) is a safe and non-invasive technique for the in vivo study of brain chemistry and metabolism. As such, it is highly applicable to the study of living brain tissue in psychiatric diseases. Several neuropathological and neuroimaging studies have suggested that abnormalities of the basal ganglia nuclei might be implicated in patients with obsessive compulsive disorder (OCD). In the present study, we performed proton [1H]MRS of the lenticular nuclei in 12 patients with OCD and 12 healthy normal comparison subjects. The peaks of N-acetyl-aspartate (NAA), creatine (Cr), and choline containing compounds (Cho) were measured. No differences between OCD patients and normal subjects were found in the NAA/Cr, Cho/Cr and NAA/Cho ratios. Our results suggest the normal viability of neuronal cells, as indicated by the quantification of NAA, Cr and Cho in the lenticular nuclei of patients with OCD. PMID- 10674363 TI - Spontaneous oscillations of cerebral blood flow velocity in the middle cerebral arteries of normal subjects and schizophrenic patients. AB - Although many regional cerebral blood flow (rCBF) studies of schizophrenic patients have been carried out, only a few studies have investigated real-time hemodynamic changes in schizophrenic patients. In the present study, we used long term monitoring of the middle cerebral artery (MCA) by non-invasive transcranial Doppler ultrasonography to obtain real-time CBF data in 55 schizophrenic patients and 20 normal comparison subjects. The mean blood flow velocity and pulsatility index (PI) of the MCA were not constant during long-term monitoring. They showed sinusoidal oscillations similar to those described in previous reports. The amplitude variations of these oscillations in both drug-naive and medicated schizophrenic patients were significantly decreased compared with findings in normal control subjects. The averaged PI values were found to be decreased in patients with illness durations of more than 10 years. After withdrawal of antipsychotic medication, both the amplitude variations of oscillations and the PI values in the drug-withdrawn patients were significantly decreased relative to findings in normal control subjects. Our results show a decreased adjustment ability of cerebral vessel resistance not only in neuroleptic-naive schizophrenic patients but also in patients with longer illness duration. Neuroleptics could affect the adjustment ability of vessel resistance. PMID- 10674364 TI - Reliability and validity of ratings of signal hyperintensities on MRI by visual inspection and computerised measurement. AB - Brain magnetic resonance imaging (MRI) scans on 98 elderly subjects, 62 with a diagnosis of schizophrenia and 36 healthy controls, were independently and blindly rated by two investigators using the visual rating methods of Fazekas et al. (Fazekas, F., Chawluk, J.B., Alavi, A., Hurtig, H.I., Zimmerman, R.A., 1987. MR signal abnormalities at 1.5 T in Alzheimer's dementia and normal aging. American Journal of Neuroradiology 8, 421-426) and Victoroff et al. (Victoroff, J., Mack, W.J., Grafton, S.T., Schreiber, S.S., Chui, H.C., 1994. A method to improve interrater reliability of visual inspection of brain MRI scans in dementia. Neurology 44, 2267-2276) for periventricular, deep white matter and subcortical gray matter signal hyperintensities (SHs) on T2-weighted images. The hyperintense signal volumes were measured by manual delineation of the signals on a workstation using Analyze software (computerised method). The subjects also underwent a detailed neuropsychological assessment. There was a high correlation between two experienced raters for both visual ratings, and the correspondence between the two methods was high. The inter-rater reliability for the computerised method was modest but significant, and the association between the visual and computerised methods was good except for ratings for SHs in subcortical nuclei. The Fazekas and computerised methods, and to a lesser degree the Victoroff method, had modest but significant correlations with some neuropsychological test measures. In conclusion, we did not demonstrate a clear superiority in reliability or validity for one demanding computerised method of rating SHs. Visual ratings should therefore be considered adequate for most clinical and research purposes, but such ratings should be accompanied by adequate training and the provision of standard reference images. PMID- 10674365 TI - Summary of the Third International Research Workshop on Alopecia Areata. PMID- 10674366 TI - Spontaneous alopecia areata-like hair loss in one congenic and seven inbred laboratory mouse strains. AB - Alopecia areata (AA) research has been hampered by the lack of suitable animal models for use in experimental procedures. AA-like hair loss has been observed in several species, including dogs, cats, horses, cattle, and nonhuman primates; however, these examples are isolated cases in outbred species of large size, limiting their use in AA research. Inbred rodent strains are ideal research models. C3H/HeJ mice can develop spontaneous AA-like hair loss and have previously been advanced as a suitable experimental model. The search for additional mouse strains with AA-like hair loss has continued. Nonscarring, inflammatory, spontaneously reversible hair loss has been observed in individual mice from several inbred mouse strains. Aside from C3H/HeJ mice, an AA-like phenotype has been observed in the substrain C3H/HeJBir, with an expression frequency of 5%. Up to 10% of individuals in an A/J mouse colony have been confirmed to develop patchy AA-like hair loss. Isolated examples of AA have also been identified in C3H/HeN/J mice, C3H/OuJ mice, HRS/J+/hr heterozygous normal mice, CBA/CaHN-Btk(xid)/J mice, and BALB.2R-H2h2/Lil mice, each with a colony frequency of less than 1%. BALB.2R-H2h2/Lil mice may also have severe nail defects. AA is regarded as rare in nonhuman species; however, nonscarring inflammatory based alopecia has been identified in several mouse strains. These examples may represent different subtypes of the heterogeneous AA phenotype. Pathologic and genetic analysis of different AA affected mouse strains may contribute to understanding AA pathogenesis and elucidating susceptibility genes. PMID- 10674367 TI - Alopecia areata is a T-lymphocyte mediated autoimmune disease: lesional human T lymphocytes transfer alopecia areata to human skin grafts on SCID mice. AB - Much evidence suggests that alopecia areata is a tissue restricted autoimmune disease. Alopecia areata responds to immunosuppressive agents, and is associated with other tissue restricted autoimmune diseases, including autoimmune thyroiditis and vitiligo. Furthermore, hair regrows when involved scalp is transplanted to nude mice. This study was undertaken to determine whether alopecia areata is mediated by T lymphocytes. Involved scalp from alopecia areata patients was grafted onto SCID mice. Additional biopsies from lesional scalp of the same patients were used to isolate T lymphocytes. These T lymphocytes were cultured with hair follicle homogenate, as well as autologous antigen presenting cells. The T lymphocytes were then injected into autologous scalp grafts on the SCID mice, which had regrown hair. Injection of scalp T lymphocytes resulted in hair loss. Hair loss was associated with the histologic and immunochemical changes of alopecia areata, including perifollicular infiltrates of T cells, along with HLA-DR and ICAM-1 expression by the follicular epithelium. Scalp T lymphocytes that had not been cultured with hair follicle homogenate did not have this effect. Preliminary data suggests hair loss requires a collaboration between CD8+ and CD4+T cells. These studies have demonstrated that alopecia areata can be induced by the transfer of T cells that recognize a hair follicle autoantigen. PMID- 10674368 TI - Alopecia areata and universalis in the Smyth chicken model for spontaneous autoimmune vitiligo. AB - The Smyth line (SL) chicken model for spontaneous, postnatal expression of vitiligo may also show varying incidences and degrees of severity ranging from alopecia areata-like to universalis-like integumental changes. Although human vitiligo patients are known to have a four times greater chance of having alopecia areata than do people without vitiligo, in the SL model, feather loss is limited to birds that show some degree of amelanosis of feather and skin tissue. Both the vitiligo and the alopecia have an autoimmune component, as shown by histologic and immunologic studies, including the correctional influences of corticosterone and cyclosporine-A. The major histocompatibility haplotype (MHC) has a major effect on the incidence and expression of the vitiligo, as well as the alopecia that occurs within vitiliginous birds. Three different MHC haplotypes were identified in the original line that was selected for vitiligo, and from these, three sublines were developed, each homozygous for a different haplotype. Of the three sublines (SL101, SL102, and SL103) the vitiligo has a significantly earlier onset and severity in the SL101 than in the other two lines. The incidence of alopecia, however, is significantly lower in the SL101 subline than in the other two. Inheritance of the vitiligo is polygenic with an additional genetic component for the alopecia trait. It is hypothesized, but as yet unproven, that a feather development defect interacts with the SL melanization and immunologic defects to initiate the partial (areata) and complete (universalis) alopecias. The alopecia universalis is rarely seen until adulthood and is characterized by short (<0.5 cm), undeveloped feathers. If feather growth resumes in these birds, the feathers dry up, cease to grow, and often break off. PMID- 10674369 TI - The genetic basis of alopecia areata: HLA associations with patchy alopecia areata versus alopecia totalis and alopecia universalis. AB - Many diseases, notably those having a strong autoimmune component, have been shown to have an association with specific human leukocyte antigens (HLA). The molecular basis for this genetic association with disease is the fact that HLA bind and present peptides derived from self and foreign protein antigens to the immune system for recognition and activation of the immune response. Previous studies with heterogeneous groups of alopecia areata (AA) patients have suggested associations with some HLA class I and class II antigens. For this study we selected only patients with long-standing disease and stratified them into two groups by strict definitions of duration and extent of disease: those with patchy AA and those with either alopecia totalis (AT) or alopecia universalis (AU). The patients were tissue typed for HLA class II antigens by biomolecular methods that provided antigen discrimination at an allele level. More than 80% of all of the AA patients typed were positive for the antigen DQB1*03 (DQ3), suggesting that this antigen is a marker for general susceptibility to AA. In addition, two other antigens were found significantly increased in frequency only in the group of AT/AU patients, DRB1*0401 (DR4) and DQB1*0301(DQ7). This strongly suggests that the two clinical types of AA, namely patchy AA versus AT/AU, can be distinguished by a genetically based predisposition to extent of disease. PMID- 10674370 TI - Alopecia areata in families: association with the HLA locus. AB - Alopecia areata (AA) is a T cell mediated disease directed against hair follicles that results in bald patches. It can range in severity from patchy (AA), to total scalp hair loss (alopecia totalis; AT) or body hair loss (alopecia universalis; AU). We have previously shown that HLA-DR4 and DR11 as well as HLA-DQ*03 alleles are increased in unrelated AA patients compared with controls. To study whether class II HLA alleles are linked to AA, we investigated 81 extended families that included 192 AA patients, including 89 with AT or AU. We also performed the transmission disequilibrium test (TDT) in 143 nuclear families. Results showed an association between alleles of HLA-DQB (p = 0.014) and HLA-DR (p = 0.010). We also performed linkage analysis in 75 families whose members' genomic DNA were available for HLA typing. Results from this analysis support linkage between AA and class II loci with a maximal LOD score of 2.42 to HLA-DQB at 5% recombination, and with a maximal LOD score of 2.34 to HLA-DR at 0% recombination. There was an increased incidence of atopic dermatitis and autoimmune thyroiditis in families. AA appears to be a class II HLA restricted organ specific immune response to the hair follicle. PMID- 10674371 TI - T cell repertoire in mice with alopecia areata. AB - It has become clear that skin infiltrating autoreactive CD4+ T helper cells play a crucial role in the initiation of alopecia areata. However, the natures of the pathogenic T cell clones as well as of the skin antigen they recognize remain obscure. Here, we analyzed the T cell receptor repertoire expressed in the spleen of diseased mice. We consistently observed the dominant expansion of a limited set of T cell clones expressing Vbeta8.2/Jbeta2.5 T cell receptor rearrangement. We conclude that T cell response in mice alopecia areata is markedly oligoclonal; a feature that may permit the design of selective immunotherapy. PMID- 10674372 TI - Immunology of the hair follicle: a short journey into terra incognita. AB - This paper delineates briefly why the immunology of the hair follicle matters (e.g., anti-infection defense, hair growth control by immunomodulatory agents, sequestration of follicular autoantigens), and which open key questions await clarification. We then focus on the murine hair follicle immune system (HIS) and its immune privilege. We show how the murine HIS is gradually constructed during hair follicle morphogenesis, and how it is transformed during hair follicle cycling. Key characteristics of the HIS are summarized, such as the absence of MHC class I expression in the anagen hair bulb and the very restricted distribution of antigen-presenting cells and intraepithelial T cells to the distal outer root sheath, which also expresses nonclassical MHC class Ib molecules. The interconnections between the HIS and the skin immune system (SIS) and potential hair growth-modulatory roles of mast cells and macrophages are addressed, and very recent findings on the human HIS are summarized. The paper closes by sketching immunobiologic, clinical, and pharmacologic perspectives in trichoimmunology that deserve the attention of immunologists, dermatologists, and hair biologists alike. PMID- 10674373 TI - The potential role of cytokines and T cells in alopecia areata. AB - T cells play an important role in alopecia areata (AA) because AA can be reinduced by the injection of hair follicle-specific CD8+ T cells into AA scalp biopsies, which were grafted onto scid mice, and the depletion of CD8+ T cells restores hair growth in the Dundee experimental bald rat. Moreover, AA can be transferred by grafting of alopecic skin from C3H/HeJ mice with AA-like hair loss onto unaffected littermates, but the onset of AA is inhibited by i.p. injection of anti-CD44v10 antibodies. Interestingly, grafted anti-CD44v10-treated mice have decreased numbers of CD8+ T cells within the skin. Beside T cells several clinical and experimental data point towards cytokines that might be crucial inducers of hair loss in AA. An aberrant expression of cytokines of the Thl type and IL-1beta has been detected in scalp areas involved by AA, and polymorphisms of cytokine genes such as IL-1-receptor antagonist, IL-1alpha, and TNF-alpha have been shown to determine disease susceptibility and severity. Moreover, IL-1 has been shown to be a potent inhibitor of human hair growth in vitro. Such IL-1 incubated hair follicles show ultrastructural changes similar to those observable in vivo. On the other hand mice transgenic for IL-1alpha develop patchy hair loss and during the depilation-induced hair cycle in C57/BL6 mice, members of the IL-1 family are overexpressed with the onset of spontaneous catagen. Taking all of the presently available data together, we may hypothesize that CD8+ T cells are of crucial importance in AA by their interaction with MHC-I restricted autoantigens, and cytolysis of their target cells. Hair loss, however, may occur because proinflammatory cytokines may interfere with the hair cycle leading to premature arrest of hair cycling. PMID- 10674374 TI - Alopecia areata-like hair loss in C3H/HeJ mice and DEBR rats can be reversed using topical diphencyprone. AB - This study demonstrates the ability to treat successfully alopecia areata-like hair loss in both mouse and rat models using topical immunotherapy with diphencyprone. PMID- 10674375 TI - The molecular basis of congenital atrichia in humans and mice: mutations in the hairless gene. AB - Congenital atrichia is a form of total alopecia inherited in an autosomal recessive pattern. In individuals affected with this form of hair loss, hairs are typically absent from the scalp, and patients are nearly completely devoid of eyebrows, eyelashes, axillary and pubic hair, following shedding of the natural hair shortly after birth. We have recently linked this disorder to the chromosomal region 8p12, and cloned the human hairless gene, which resides within this interval. We have identified several mutations in the hairless gene in atrichia families from around the world. In hairless mice, the hair matrix cells appear to undergo a premature and massive apoptosis, together with a concomitant decline in Bcl-2 expression, a loss of NCAM positivity, and a disconnection with the overlying epithelial sheath essential for the movement of the dermal papilla. As a consequence, the hair bulb and dermal papilla remain stranded in the dermis, and indispensible messages between the dermal papilla and stem cells in the bulge are not transmitted, so no further hair growth occurs. These findings suggest that the hairless gene product may play a crucial role in maintaining the delicate balance between cell proliferation, differentiation and apoptosis in the hair follicle, as well as in the interfollicular epidermis. PMID- 10674376 TI - Hair defects in Hoxc13 mutant mice. AB - Hox genes encode transcription factors that are important during normal embryonic development of diverse organisms including vertebrates. In mammals, Hox genes are responsible for conferring regional identity in embryonic tissues, including the limb bud, the neural tube, the presomitic mesoderm and the intestinal tract. Recent studies have demonstrated expression of Hox genes in skin and hair follicles, suggesting potential functions for these genes in epidermal appendages. These studies are reviewed here with emphasis on Hoxc13, as Hoxc13 mutants are the first Hox mutants to demonstrate overt hair defects. In addition, because Hoxc13 does not show regionally restricted expression in the skin, as demonstrated for other Hox genes, the potentially different roles of Hoxc13 versus other Hox genes in the skin are discussed. PMID- 10674377 TI - The presence of loose anagen hairs obtained by hair pull in the normal population. AB - To help determine the specificity of "loose anagen" (LA) hairs in Loose Anagen Syndrome, the presence or absence of LA hairs on a gentle but firm hair pull was evaluated in 110 normal subjects from a 0.5 to 83 y old. In children < or =10 y old, 61% had LA hairs on hair pull evaluation and 73% of all hairs obtained were LA hairs. In contrast, LA hairs were found in only two of 87 (2%) normal postpubescent subjects. The number of LA hairs was small in normal children (1-2 per hair pull) and a maximum of one out of every 6-7 hair pulls in adults, far less than that reported with Loose Anagen Syndrome. Although the mere presence of LA hairs on a hair pull test is thus not specific for LAS in children, the number per hair pull may have diagnostic significance. Correlation of these findings with the various hair disorder phenotypes currently termed Loose Anagen Syndrome will be important. PMID- 10674378 TI - Marie Unna congenital hypotrichosis: clinical description, histopathology, scanning electron microscopy of a previously unreported large pedigree. AB - Marie Unna congenital hypotrichosis (MUCH) is a rare autosomal dominant condition in which abnormalities are confined to hair shaft structure and hair density. We report a six-generation pedigree consisting of 59 members of whom 16 are affected; nine identified affected individuals are living. Affected individuals are born with adequate, normal to coarse hair. During early infancy the scalp hair becomes more coarse and wiry and stands out from the head. All affected individuals have sparse to absent eyebrows, eyelashes and body hair including secondary sexual hair. In some individuals, scalp hair is progressively lost beginning at puberty or beyond, until only a sparse fringe in the tonsorial distribution remains. The hair shafts are uniformly increased in diameter, measuring up to 0.12 mm. Individual hair shafts are deeply pigmented, variable in diameter, twisted, and bent at odd angles; some have a longitudinal groove visible on scanning electron microscopy. Cross-sectional shapes are variable and irregular, exhibiting oval, angular to reniform shapes. Multiple anagen hairs are extractable on gentle hair pull. Other ectodermal structures are unaffected except for exceptionally widely spaced upper incisor teeth seen in 50% of affected individuals. Histologically, there are dramatically reduced numbers of follicles per unit area, averaging nine total hairs per 4 mm cross-section as compared with a normal of 40. A mild to moderate inflammatory infiltrate is present, but little fibrosis and no scarring. The mechanism of progressive hair loss is unknown. PMID- 10674379 TI - Trichodysplasia spinulosa--a newly described folliculocentric viral infection in an immunocompromised host. AB - This is a case report of an immunocompromised individual who presented with progressive alopecia, friable follicular spinous processes, and erythematous, indurated papules. Examination of skin biopsies using light microscopy and immunohistochemistry revealed pathologic changes of the follicular inner root sheath epithelium with dystrophic trichohyaline granules. Electron microscopy of thin sections of tissue revealed intracellular viral particles with a size and appearance consistent with those in the Papovaviridae family. Electron microscopy of negatively stained extract from a homogenized lesion also demonstrated icosahedral viruses with papovavirus morphology. We believe this is a previously unreported folliculocentric viral infection in an immunosuppressed human host and have termed this entity "trichodysplasia spinulosa". PMID- 10674380 TI - Hair follicle apoptosis and Bcl-2. AB - Hair follicle (HF) morphogenesis and cycling are characterized by a tightly controlled balance of proliferation, differentiation and apoptosis. The members of the bcl-2 family of proto-oncogenes are important key players in the apoptosis control machinery of most cell types. Bcl-2, an apoptosis inhibitor, and Bax, an apoptosis promoter, show tightly regulated, hair cycle-dependent expression patterns: during catagen, the distal ORS of the HF remains strongly positive for Bcl-2 and Bax; in contrast, the proximal epithelial part of the HF loses most Bcl 2 expression while it remains strongly positive for Bax. In Bcl-2 null mice, skin becomes markedly hypopigmented during the first postnatal anagen probably due to increased melanocyte apoptosis. Reportedly, these mice also show a retardation of the first anagen development after birth. Transgenic mice overexpressing Bcl-2 under the control of the keratin-1 promoter display multifocal epidermal hyperplasia and aberrant expression of keratin-6, while alterations of HF cycling have not been investigated. Surprisingly, Bcl-2 overexpression under the control of the keratin-14 promoter leads to accelerated catagen progression and increased chemotherapy-induced apoptosis, HF dystrophy and alopecia. Transgenic mice overexpressing Bcl-X(L), another anti-apoptotic bcl-2 family member, under the control of the K14 promoter, reportedly also display accelerated catagen development. These and other Bcl-2 transgenic and null mice are now available to further dissect the as yet unclear, and likely complex, role of Bcl-2 in HF growth and pigmentation. PMID- 10674381 TI - Msx-2 and the regulation of organ size: epidermal thickness and hair length. AB - During organogenesis, the issue of size regulation is as important as shape and differentiation. We propose that the regulation of the dimensions of the epithelium and its appendages (length, width, thickness) are based on regulation of cell numbers in specific sites, reflecting the input and output of cells in that region. This process is in turn regulated by the flow from the domain of proliferating cells to the domain of postmitotic differentiated cells. When the homeobox gene Msx-2 is over-expressed in transgenic mice under the control of the CMV promoter, the epidermis is thickened with hyperproliferation and hyperkeratosis. Hairs are shorter and the matrix region is shrunken. We suggest that Msx-2 may be one of the regulators involved in the control of organ size, and the above phenotypes are the manifestations of an increased cellular flow from proliferation domain to differentiation domain in the tissue. PMID- 10674382 TI - Measuring reversal of hair miniaturization in androgenetic alopecia by follicular counts in horizontal sections of serial scalp biopsies: results of finasteride 1 mg treatment of men and postmenopausal women. AB - Hair regrowth was evaluated by histologic analysis in men and women treated for androgenetic alopecia, by counting follicles in horizontal sections of scalp biopsies. Serial 4mm punch biopsies were taken at baseline and after 12mo of treatment from the transitional area of hair thinning between normal hair and vertex balding in men, and in an area of frontal/parietal thinning in women. Horizontal sections of reticular and papillary dermis were read by one observer, blinded to patient, treatment, and time. All terminal hair bulbs, terminal anagen and telogen hairs, and vellus and vellus-like miniaturized hairs were counted. Twenty-six men aged 18-41y, comprising 14 on finasteride 1 mg daily and 12 on placebo, and 94 postmenopausal women, aged 41-60y, comprising 44 on finasteride 1 mg daily and 50 on placebo, were evaluated. In the male study, the terminal hairs increased from a mean baseline count of 15.5-20.9 after 12mo of finasteride, versus 17.3-18.3 in the placebo patients. The miniaturized hairs decreased from 26.7 to 23.6 with finasteride versus 21.3-20.3 with placebo. The terminal-to vellus ratio increased more in the finasteride than in the placebo patients, suggesting some reversal of the miniaturization process with finasteride. In the female study, no significant differences in follicular counts were found between the finasteride and placebo groups after 12mo of treatment. Follicular counts in horizontal sections provide an informative adjunct to noninvasive measures used in hair growth studies. Finasteride appears to be capable of reversing hair miniaturization in androgenetic alopecia in young to middle-aged men, but not in postmenopausal women. PMID- 10674383 TI - Effects of 17-beta-estradiol and ICI 182 780 on hair growth in various strains of mice. AB - 17-beta-Estradiol (10 nmol per 200 microl acetone) applied topically twice weekly to the clipped dorsal surface of C57BL/6 or C3H female mouse skin prevented hair growth, as previously described in the CD-1 mouse strain. Twice weekly topical application of the estrogen receptor antagonist, ICI 182 780 (10nmol per 200microl acetone), induced the telogenanagen transition and produced early pigmentation appearance in skin and hair growth in C57BL/6 and C3H female mice. Whereas twice weekly topical application of 10nmol 17-beta-estradiol blocked hair growth, the intraperitoneal administration of this dose twice weekly did not block hair growth, suggesting a direct cutaneous effect of 17-beta-estradiol. We also evaluated the effect of 17-alpha-estradiol, 17-beta-estradiol, and ICI 182 780 on hair growth in male mice. As observed in female mice, 17-beta-estradiol was a potent inhibitor of hair growth and ICI 182 780 stimulated hair growth; however, unlike the results previously observed in female mice, 17-alpha estradiol was a potent inhibitor of hair growth in male mice. These results demonstrate that (i) the route of administration of 17-beta-estradiol is critical for its ability to block hair growth; (ii) C57BL/6 and C3H mice, two commonly employed mouse strains for hair growth studies, responded to 17-beta-estradiol and ICI 182 780 in a manner similar to that described in CD-1 mice; and (iii) the hair follicles of male and female mice respond similarly to 17-beta-estradiol and ICI 182 780, but display striking sex differences in the response to 17-alpha estradiol on hair growth. PMID- 10674384 TI - In vitro main pathways of steroid action in cultured hair follicle cells: vascular approach. AB - The known role of steroids on the hair follicle leads us to investigate their effects on hair follicle cell angiogenic responses in vitro. We verified, using the immunohistochemical technique, whether human occipital scalp follicle cells express steroid receptors in vitro. We showed that androgen and estrogen receptors were expressed by dermal papilla cells (DPC) and keratinocytes from the outer root sheath in vitro. With regard to steroidal enzymes (type I and II 5alpha-reductases and Cytochrome-p-450-aromatase), the type I 5alpha-reductase gene is much more expressed in DPC than in dermal fibroblasts; however, the type II 5a-reductase gene is transcribed more in dermal fibroblasts than in DPC. The transcription of the two 5alpha-reductase isoform genes in cultured DPC is regulated by a 5alpha-reductase inhibitor. We also demonstrated that DPC, dermal fibroblasts, and outer root shealth keratinocytes expressed cytochrome-p-450 aromatase. Using ELISA and reverse transcriptase-polymerase chain reaction, we investigated the role played by some steroids (estrogens, androgens, antiandrogens) in the modulation of vascular endothelial growth factor (VEGF) expression by DPC. The association of different treatments of DPC (5alpha reductase inhibitor and androgen receptor antagonist) shows a great stimulation of VEGF and aromatase expression. Strong stimulation of VEGF protein and gene expression is observed in the presence of 17beta-estradiol. Also, the concentration-dependent inhibition of VEGF expression by DPC using the cytochrome p-450-aromatase inhibitor, confirms the involvement of this estrogenic pathway in the regulation of VEGF expression in vitro. PMID- 10674385 TI - Human hair follicle bulge cells are biochemically distinct and possess an epithelial stem cell phenotype. AB - Stem cells are vital for the homeostasis of self-renewing tissues and their manipulation may have wide ranging applications, including gene therapy, wound repair, and tissue transplantation. Although rodent hair follicle stem cells have been localized to the follicle bulge, the location of human hair follicle stem cells is less clear, and their characterization has been hampered by a lack of cellular markers for the bulge area. We demonstrate that the C8/144B monoclonal antibody, originally raised against a CD8 peptide sequence, immunostains the human hair follicle bulge. We show that this antibody recognizes cytokeratin 15 (K15) in keratinocytes, and that K15-positive bulge cells possess a stem cell phenotype characterized by their slowly cycling nature, proliferation at the onset of new hair follicle growth, and high level of beta1 integrin expression. These results localize human hair follicle stem cells to the bulge and suggest that K15 is preferentially expressed in epithelial stem cells. PMID- 10674386 TI - Early events in skin appendage formation: induction of epithelial placodes and condensation of dermal mesenchyme. AB - The formation of skin appendages represents a morphogenetic process through which a homogeneous system is converted into a patterned system. We have pursued molecules involved in the early placode induction and mesenchymal condensation stages of this process. We found that intracellular and extracellular signaling molecules collaborate to position the location of feather primordia and initiate mesenchymal condensations mediated by adhesion molecules. During the inductive stage, cells interact in a fashion best described by a reaction-diffusion mechanism. Thus in early feather morphogenesis, low level adhesion molecules drive cell interactions. The interactions were modulated by extracellular signaling molecules, which eventually increase the level of signaling molecules at sites of feather initiation and subsequently the level of adhesion molecules (Jiang et al, 1999a). These physico-chemical events lead to the formation of dermal condensations and epithelial placodes at sites of feather primordia, thus achieving the earliest and most fundamental events of skin appendage formation: induction. PMID- 10674387 TI - Phenotypic determination of epithelial appendages: genes, developmental pathways, and evolution. AB - Epithelial appendages are derivatives of epithelia that elaborate to form specialized structures and functions. The appendage can protrude out, such as in teeth and feathers, or invaginate in, such as in glands. The epithelia can be ectodermal, such as in hairs, or endodermal, such as in livers. Using feather as a prototype of epithelial appendage, we study the molecular signals involved in the successive stages of epithelial-mesenchymal interactions during morphogenesis. We propose that these form the basics of gene networks, which can be integrated to gene supernetwork and totinetwork. Because the unit of development is molecular pathway rather than single molecule, and the unit of morphogenesis is cell group rather than single cell, we make the analogy between genes/developmental pathways and words/sentences. The study of developmental pathways in epithelial appendage organogenesis will help us to understand the grammar of genes and the basic rules in constructing regulated new growth. This knowledge may contribute to the study of cancer biology (deregulated new growth) and organ regeneration. PMID- 10674388 TI - Hair cycle-dependent expression of hepatocyte growth factor (HGF) activator, other proteinases, and proteinase inhibitors correlates with the expression of HGF in rat hair follicles. AB - We previously reported that hepatocyte growth factor (HGF) had a stimulatory effect on hair growth in vivo and in vitro. The secreted inactive form of HGF is processed into an active form by serine proteinases such as HGF activator and urokinase. The mRNA expressions of various proteinases and their inhibitors in relation to HGF activation in hair growth were examined using animals with a synchronous hair cycle. Total RNA were extracted from the anterior dorsal skin of rats in different hair cycle stages, and mRNA expressions of the specific genes were compared using semiquantitative reverse transcription polymerase chain reaction. The mRNA of HGF, HGF activator, urokinase, plasminogen activator inhibitor (PAI)-1, nexin-1, matrix metalloproteinase (MMP)-2, and tissue inhibitor of metalloproteinase (TIMP)-1 were expressed strongly in anagen tissue and slightly in telogen tissue. Moreover, topical application of 1% minoxidil sulfate to the anterior dorsal skin of rats in telogen stimulated hair growth and increased the mRNA expressions of HGF and MMP-2. These findings suggest that some proteinases and their inhibitors, strongly expressed in anagen, may act as hair growth regulatory molecules, and may also be involved in processing the latent form of HGF. PMID- 10674389 TI - Peribulbar innervation and substance P expression following nonpermanent injury to the human scalp hair follicle. AB - The hair pluck procedure alters the anatomy of the anagen hair bulb. Hemorrhage can occur in the mesenchymal sheath and breaks at the proximal epithelium, above or around the upper third of the dermal papilla, have been reported. We hypothesized that innervation, as identified with protein gene product 9.5 (PGP 9.5), and expression of the neuropeptide Substance P (SP) within the dermal papilla would also be altered following plucking. We focused on studying SP as this neuropeptide has been associated with several cellular responses, including anagen hair growth in the C57BL/6 mouse model. Four millimeter punch biopsies were obtained from the occipital scalp of two healthy adults. Hair was then plucked and additional biopsies were obtained immediately, and at 1 d, 1 wk, and 1 mo after plucking. Each set was processed for immunohistochemical analyses and in-focus optical sections of the dermal papilla were captured by laser scanning confocal microscopy and later reconstructed into single images. Following injury, SP was expressed in a disorganized pattern below the dermal papilla. There was also a significant reduction in labeled neuronal cells, and SP expression was enhanced within peribulbar blood vessels at 1 d and 1 wk. By 1 mo, peribulbar nerves, vessels, and SP expression were similar to baseline observations. It remains to be ascertained whether PGP 9.5, also known as unbiquitin hydrolase, and SP are involved in the proliferation of new matrix cells in the human scalp hair follicle following injury. PMID- 10674390 TI - Identification of a novel SCD gene and expression of the SCD gene family in mouse skin. AB - We have refined the position of asebia locus by genotyping DNA from more than 600 backcross mice derived from asebia mouse and a genetically unrelated strain. One of the candidate genes in the locus is stearoyl-CoA desaturase (SCD). Previously two members of this gene family, namely SCD1 and SCD2, have been described. We have found, for the first time, that these SCD genes are expressed in skin. Moreover, we have identified a third species of SCD in the mouse skin. The most prominent SCD species is SCD1 in the mouse skin. The implications of this gene family to skin are discussed. PMID- 10674391 TI - The fate of hair follicle melanocytes during the hair growth cycle. AB - The fate of the follicular pigmentary unit during the hair growth cycle has long been one of the great enigmas of both hair follicle and pigment cell biology. Although melanocytes are distributed in several different compartments of the anagen hair follicle, melanogenically active cells are located only in the hair bulb, where they are directly involved in hair shaft pigmentation. These pigment cells are readily detectable only when they become melanogenically active during anagen III of the hair growth cycle. Thus, their status during hair follicle regression (catagen), when melanogenesis is switched off, until they re-appear again as pigment-producing cells in the anagen III hair follicle, has remained poorly defined. Historically, it has been proposed that hair bulb melanocytes adopt a self-perpetuating, catagen-resistant strategy of de-differentiation during hair follicle regression and re-differentiation upon entry into a new anagen phase; however, this explanation remains problematic in the absence of evidence for de-differentiation/re-differentiation plasticity in most nonmalignant cell systems. PMID- 10674392 TI - Avian integument provides multiple possibilities to analyse different phases of skin appendage morphogenesis. AB - To analyse the morphogenic events during skin appendage formation, it is important to have an animal model that offers distinct patterns at various stages of development and is accessible to analysis using state of the art technology. The avian integument is such a model. Combining experimental embryologic approaches, organ cultures, and gene transduction technology, we are now able to begin to address the molecular basis of pattern formation, primordium initiation, anterior-posterior axis formation, proximo-distal axis formation, phenotypic determination, and others. Parallel mechanisms are usually found in feathers and hairs, and the avian integument model has matured to be a major source of new findings in the study of skin appendage morphogenesis. More information on the avian integument model can be found at website http://www.hsc.usc.educmchuong. PMID- 10674393 TI - Chronobiology of the hair follicle: hunting the " hair cycle clock". AB - The hair follicle (HF) is the only mammalian organ that undergoes life-long, cyclic transformations from long stages of growth (anagen), via rapid, apoptosis driven organ involution (catagen) to a stage of relative "resting" (telogen). The controls that underlie these transformations clearly reside in and/or around the HF itself, and are likely to reflect - essentially autonomous, yet highly manipulable - changes in the local signalling milieu of e.g., hair growth modulatory growth factors, cytokines, hormones and adhesion molecules. Yet the molecular nature and organization of the "hair cycle clock" (HCC) that drives these cyclic switches in the local signalling milieu remain obscure, and there is not even a fully satisfactory theory of hair cycle control. Since deciphering of the HCC is of paramount clinical importance, and since corresponding working hypotheses are badly needed to guide the design of more incisive experiments that identify the elusive central "oscillator" mechanism behind the HCC, we discuss basic requirements any convincing HCC theory should meet. After arguing that at least four distinct timing devices underlie HF chronobiology ("morphogenesis clock", "cycling inducer", "desynchronizer", and the actual HCC), previously proposed HCC theories are briefly and critically reviewed. In the light of intriguing regulatory similarities between the HCC and the cell cycle machinery, we suggest here that the HCC may be driven by autonomous, cell cycle-coupled secretory activities of the HF mesenchyme, namely by changes in the G0/G1 associated secretion of "papilla morphogens" by dermal papilla fibroblasts. Hopefully, this provocative hypothesis will encourage the proposition of novel, comprehensive HCC theories. PMID- 10674394 TI - Silencing subdomains of v-ErbA interact cooperatively with corepressors: involvement of helices 5/6. AB - Members of the thyroid hormone receptor (TR) family act on vertebrate development and homeostasis by activating or repressing transcription of specific target genes in a ligand-dependent way. Repression by TR in the absence of ligand is mediated by an active silencing mechanism. The oncogene v-ErbA is a variant form of TR unable to bind hormone and thus acts as a constitutive repressor. Functional studies and mutation analysis revealed that the TR/v-ErbA silencing domain is composed of three silencing subdomains (SSD1-3) which, although nonfunctional individually, synergize such that silencing activity is restored when they are combined in a heteromeric complex. Here we demonstrate, using protein interaction assays in vitro and in vivo, that the inactive v-ErbA point mutant L489R within helix 5/6 in SSD2 fails to interact with the two corepressors N-CoR (nuclear receptor corepressor) or SMRT (silencing mediator of retinoic acid and thyroid hormone receptor). Furthermore, mutants in SSD1 and SSD3 exhibit a reduced corepressor recruitment corresponding to their weak residual silencing activity. In mammalian two-hybrid assays, only the combination of all three silencing subdomains, SSD1-3, leads to a cooperative binding to the corepressors N-CoR or SMRT comparable to that of the full-length v-ErbA repression domain. In conclusion, full silencing activity requires corepressor interaction with all three silencing subdomains, SSD1-3. Among these, SSD2 is a new target for N-CoR and SMRT and is essential for corepressor binding and function. PMID- 10674395 TI - Transcription factors Oct-1 and C/EBPbeta (CCAAT/enhancer-binding protein-beta) are involved in the glutamate/nitric oxide/cyclic-guanosine 5'-monophosphate mediated repression of mediated repression of gonadotropin-releasing hormone gene expression. AB - The physiological actions of nitric oxide (NO) as a signaling molecule in endothelial and brain cells and as a toxic molecule used by activated immune cells have been the focus of a wide range of studies. Nevertheless, the downstream effector molecules of this important neuromodulator are not well understood. We have previously demonstrated that expression of the gene for the reproductive neuropeptide, GnRH, is repressed by the glutamate/NO/cyclic GMP (cGMP) signal transduction pathway through cGMP-dependent protein kinase in the hypothalamic GnRH-secreting neuronal cell line GT1-7. This repression localized within a previously characterized 300-bp neuron-specific enhancer. Here, we find that mutation of either of two adjacent elements within the enhancer eliminates repression by this pathway. An AT-rich sequence located at -1695 has homology to the octamer motif known to bind POU-homeodomain proteins, while the adjacent element at -1676 has homology to the C/EBP (CCAAT/enhancer-binding protein) protein family consensus sequence. Antibody supershift assays reveal that one of the proteins bound at the -1695 sequence is Oct-1, and one of the proteins bound to the element at -1676 is C/EBPbeta. These two proteins can bind simultaneously to the adjacent -1695 and -1676 binding sites in vitro. In nuclear extracts of GT1-7 cells treated with an NO donor, the intensity of the Oct-1 complex is increased. However, although Western blot analysis indicates that neither Oct-1 nor C/EBPbeta protein levels are increased, the relative binding affinity of Oct 1 is increased. Dephosphorylation of the nuclear extracts decreases binding of the Oct-1 complex to the -1695 site only in NO donor-treated extracts. Thus, we conclude that Oct-1 and C/EBPbeta are both downstream transcriptional regulators involved in the repression of GnRH gene expression by the glutamate/NO/ cGMP signal transduction pathway. PMID- 10674396 TI - Multiple signaling pathways mediate interleukin-4-induced 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase type 1 gene expression in human breast cancer cells. AB - The 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase (3beta-HSD) isoenzymes catalyze an essential step in the formation of all classes of active steroid hormones. We have recently shown that 3beta-HSD type 1 gene expression is specifically induced by interleukin (IL)-4 and IL-13 in breast human cancer cell lines and in normal human mammary epithelial cells in primary culture. There is evidence that IL-4 stimulates bifurcating signaling pathways in which the signal transducer and activator of transcription-6 (Stat6)-signal pathway is involved in differentiation and gene regulation, whereas insulin receptor substrate (IRS) proteins mediate the mitogenic action of IL-4. In fact, we have shown that Stat6 was activated by IL-4 in all cell lines studied where IL-4 induced 3beta-HSD expression, but not in those that failed to respond to IL-4. The present study was designed to investigate the potential contribution of IRS proteins and their downstream targets to IL-4-induced 3beta-HSD type 1 gene expression. IL-4 rapidly induced IRS-1 and IRS-2 phosphorylation in ZR-75-1 human breast cancer cell lines. Moreover, insulin-like growth factor (IGF)-I and insulin, which are well known to cause IRS-1 and IRS-2 phosphorylation, increased the stimulatory effect of IL-4 on 3beta-HSD activity. IRS-1 and IRS-2 are adapter molecules that provide docking sites for different SH2-domain-containing proteins such as the phosphatidylinositol (PI) 3-kinase. In this light, the inhibition of IL-4-induced 3beta-HSD expression by wortmannin and LY294002, two potent PI 3-kinase inhibitors, indicates the probable involvement of the PI 3-kinase signaling molecules in this response to IL-4. Furthermore, it has been suggested that the IRS proteins are part of the signaling complexes that lead to activation of the mitogen-activated protein (MAP) kinase by insulin; thus we investigated the potential role of the MAP kinase (MAPK) cascade in the IL-4 action. In ZR-75-1 cells, both the activation of MAPK by IL-4 and the IL-4-induced 3beta-HSD activity were completely blocked by PD98059, an inhibitor of MAPK activation. Wortmannin also blocked MAPK activation by IL-4, IGF-I, and insulin, suggesting that the MAPK cascade acts as a downstream effector of PI 3-kinases. To further understand the cross-talk between signaling pathways involved in IL-4 action, we investigated the possible involvement of protein kinase C (PKC). The potential role of PKC was suggested by the observation that the well known PKC activator phorbol-12-myristate-13-acetate (PMA) potentiated the IL-4-induced 3beta-HSD activity. Taken together, these findings suggest the existence of a novel mechanism of gene regulation by IL-4. This mechanism would involved the phosphorylation of IRS-1 and IRS-2, which transduce the IL-4 signal through a PI 3-kinase- and MAPK-dependent signaling pathway. The inability of IGF-I, insulin, and PMA to stimulate 3beta-HSD expression by themselves in the absence of IL-4 makes obvious the absolute requirement of an IL-4-specific signaling molecule. Our findings thus suggest that the multiple pathways downstream of IRS-1 and IRS 2 must act in cooperation with the IL-4-specific transcription factor Stat6 to mediate the induction of 31beta-HSD type 1 gene expression in ZR-75-1 human breast cancer cells. PMID- 10674397 TI - Apoptosis mediated by activation of the G protein-coupled receptor for parathyroid hormone (PTH)/PTH-related protein (PTHrP). AB - The present studies were carried out to evaluate the mechanisms by which PTH/PTHrP receptor (PTHR) activation influences cell viability. In 293 cells expressing recombinant PTHRs, PTH treatment markedly reduced the number of viable cells. This effect was associated with a marked apoptotic response including DNA fragmentation and the appearance of apoptotic nuclei. Similar effects were evidenced in response to serum withdrawal or to the addition of tumor necrosis factor (TNFalpha). Addition of caspase inhibitors or overexpression of bcl-2 partially abrogated apoptosis induced by serum withdrawal. Caspase inhibitors also protected cells from PTH-induced apoptosis, but overexpression of bcl-2 did not. The effects of PTH on cell number and apoptosis were neither mimicked by activators of the cAMP pathway (forskolin, isoproterenol) nor blocked by an inhibitor (H-89). However, elevation of Ca(i)2+ by addition of thapsigargin induced rapid apoptosis, and suppression of Ca(i)2+ by overexpression of the calcium- binding protein, calbindin D28k, inhibited PTH-induced apoptosis. The protein kinase C inhibitor GF 109203X partially inhibited PTH-induced apoptosis. Regulator of G protein signaling 4 (RGS4) (an inhibitor of the activity of the alpha-subunit of Gq) suppressed apoptotic signaling by the PTHR, whereas the C terminal fragment of GRK2 (an inhibitor of the activity of the beta(gamma) subunits of G proteins) was without effect. Chemical mutagenesis allowed selection of a series of 293 cell lines resistant to the apoptotic actions of PTH; a subset of these were also resistant to TNFalpha. These results suggest that 1) apoptosis produced by PTHR and TNF receptor signaling involve converging pathways; and 2) Gq-mediated phospholipase C/Ca2+ signaling, rather than Gs mediated cAMP signaling, is required for the apoptotic effects of PTHR activation. PMID- 10674398 TI - Dual regulation of somatostatin receptor subtype 1 gene expression by pit-1 in anterior pituitary GH3 cells. AB - Somatostatin represents a major release inhibiting factor for hypophyseal hormones and mediates its action via five receptor subtypes, sst1-sst5, that are all present in the anterior pituitary. The pituitary specific transcription factor Pit-1 is essential for the pituitary development and pituitary-specific gene expression. Here the transcriptional regulation of the sst1 gene, which contains putative Pit-1-binding sites, was studied in anterior pituitary GH3 cells. We found that a fragment of 2 kb suffices to drive the expression of a reporter gene specifically in this cell line. Positive and negative cis regulatory elements contributed to the promoter activity. Among these elements two functional binding sites for Pit-1 were identified. While the proximal site mediated transcriptional activation, the distal site attenuated transcription of reporter gene constructs. Mutations of the proximal Pit-1 site prevented expression of the reporter gene. Targeting Pit-1 mRNA by antisense oligonucleotides caused inhibition of transcription of reporter gene constructs containing the proximal Pit-1-binding site. Moreover, the expression of the endogenous sst1 gene in GH3 anterior pituitary cells was blocked. This resulted in reduced sst1 levels at the plasma membrane. Reduced sst1 levels were associated with a diminished antisecretory response to the sst1-specific agonist CH-275 and somatostatin. These results demonstrate the importance of Pit-1 for the expression of the sst1 gene, which hence is placed under common genetic control with the genes for hypophysiotropic hormones and the gene for the receptor of GH-releasing hormone. PMID- 10674399 TI - The nematode leucine-rich repeat-containing, G protein-coupled receptor (LGR) protein homologous to vertebrate gonadotropin and thyrotropin receptors is constitutively active in mammalian cells. AB - The receptors for LH, FSH, and TSH belong to the large G protein-coupled, seven transmembrane protein family and are unique in having a large N-terminal extracellular (ecto-) domain containing leucine-rich repeats important for interactions with the large glycoprotein hormone ligands. Recent studies indicated the evolution of an expanding family of homologous leucine-rich repeat containing, G protein-coupled receptors (LGRs), including the three known glycoprotein hormone receptors; mammalian LGR4 and LGR5; and LGRs in sea anemone, fly, and snail. We isolated nematode LGR cDNA and characterized its gene from the Caenorhabditis elegans genome. This receptor cDNA encodes 929 amino acids consisting of a signal peptide for membrane insertion, an ectodomain with nine leucine-rich repeats, a seven-TM region, and a long C-terminal tail. The nematode LGR has five potential N-linked glycosylation sites in its ectodomain and multiple consensus phosphorylation sites for protein kinase A and C in the cytoplasmic loop and C tail. The nematode receptor gene has 13 exons; its TM region and C tail, unlike mammalian glycoprotein hormone receptors, are encoded by multiple exons. Sequence alignments showed that the TM region of the nematode receptor has 30% identity and 50% similarity to the same region in mammalian glycoprotein hormone receptors. Although human 293T cells expressing the nematode LGR protein do not respond to human glycoprotein hormones, these cells exhibited major increases in basal cAMP production in the absence of ligand stimulation, reaching levels comparable to those in cells expressing a constitutively activated mutant human LH receptor found in patients with familial male-limited precocious puberty. Analysis of cAMP production mediated by chimeric receptors further indicated that the ectodomain and TM region of the nematode LGR and human LH receptor are interchangeable and the TM region of the nematode LGR is responsible for constitutive receptor activation. Thus, the identification and characterization of the nematode receptor provides the basis for understanding the evolutionary relationship of diverse LGRs and for future analysis of mechanisms underlying the activation of glycoprotein hormone receptors and related LGRs. PMID- 10674400 TI - Involvement of STAT5 (signal transducer and activator of transcription 5) and HNF 4 (hepatocyte nuclear factor 4) in the transcriptional control of the hnf6 gene by growth hormone. AB - HNF-6 is a tissue-restricted transcription factor that participates in the regulation of several genes in liver. We reported earlier that in adult rats, HNF 6 mRNA concentration in liver drops to almost undetectable levels after hypophysectomy and returns to normal after 1 week of GH treatment. We now show that this results from a rapid effect of GH, and we characterize its molecular mechanism. In hypophysectomized rats, HNF-6 mRNAs increased within 1 h after a single injection of GH. The same GH-dependent induction was reproduced on isolated hepatocytes. To determine whether GH regulates hnf6 expression at the gene level, we studied its promoter. DNA binding experiments showed that 1) the transcription factors STAT5 (signal transducer and activator of transcription 5) and HNF-4 (hepatocyte nuclear factor 4) bind to sites located around -110 and 650, respectively; and 2) STAT5 binding is induced and HNF-4 binding affinity is increased in liver within 1 h after GH injection to hypophysectomized rats. Using transfection experiments and site-directed mutagenesis, we found that STAT5 and HNF-4 stimulated transcription of an hnf6 gene promoter-reporter construct. Furthermore, GH stimulated transcription of this construct in cells that express GH receptors. Consistent with our earlier finding that HNF-6 stimulates the hnf4 and hnf3beta gene promoters, GH treatment of hypophysectomized rats increased the liver concentration of HNF-4 and HNF-3beta mRNAs. Together, these data demonstrate that GH stimulates transcription of the hnf6 gene by a mechanism involving STAT5 and HNF-4. They show that HNF-6 participates not only as an effector, but also as a target, to the regulatory network of liver transcription factors, and that several members of this network are GH regulated. PMID- 10674401 TI - Association of 2',5'-oligoadenylate synthetase with the prolactin (PRL) receptor: alteration in PRL-inducible stat1 (signal transducer and activator of transcription 1) signaling to the IRF-1 (interferon-regulatory factor 1) promoter. AB - The PRL receptor (PRL-R) signals through the Janus tyrosine kinases (JAK) and other non-JAK tyrosine kinases, some of which are preassociated with the PRL-R. To clone PRL-R interacting proteins, the intracellular domain (ICD) of the long form of the PRL-R was used in a yeast two-hybrid screen of a human B cell cDNA library. One PRL-R interacting protein was identified as the 42-kDa form of the enzyme 2',5'-oligoadenylate synthetase (OAS). The in vivo interactions in yeast were further confirmed by an in vitro interaction assay and by coimmunoprecipitation in transfected mammalian cells. Functionally, OAS reduced the basal activity of two types of promoters in transiently transfected COS-1 cells. In the presence of PRL, OAS inhibited PRL induction of the immediate early IRF-1 (interferon-regulatory factor 1) promoter, but not PRL induction of the differentiation-specific beta-casein promoter, suggesting that OAS exerts specific effects on immediate early gene promoters. The inhibitory effects of OAS were accompanied by a reduction in PRL-inducible Stat1 (signal transducer and activator of transcription 1) DNA binding activity at the IRF-1 GAS (interferon gamma-activated sequence) element. These results demonstrate a novel interaction of OAS with the PRL-R and suggest a role for OAS in modulating Stat1-mediated signaling to an immediate early gene promoter. Although previously characterized as a regulator of ribonuclease (RNase) L antiviral responses, OAS may have additional effects on cytokine receptor signal transduction pathways. PMID- 10674402 TI - Prolactin enhances CCAAT enhancer-binding protein-beta (C/EBP beta) and peroxisome proliferator-activated receptor gamma (PPAR gamma) messenger RNA expression and stimulates adipogenic conversion of NIH-3T3 cells. AB - Extracellular stimuli trigger adipocyte differentiation by inducing the complex cascades of transcription. Transcription factors CCAAT enhancer-binding proteins (C/EBPs) and peroxisome proliferator-activated receptor gamma (PPARgamma) play crucial roles in this process. Although ectopic expression of these factors in NIH-3T3 cells, a multipotential mesenchymal stem cell line, results in adipogenic conversion, little is known as to hormonal factors that regulate adipogenesis in these cells. In this report we demonstrate that PRL, a lactogenic hormone, enhances C/EBPbeta and PPARbeta mRNA expression and augments adipogenic conversion of NIH-3T3 cells. Moreover, we show that ectopic expression of the PRL receptor in NIH-3T3 cells results in efficient adipocyte conversion when stimulated with PRL and a PPARgamma ligand, as evidenced by expression of the adipocyte differentiation-specific genes as well as the presence of fat-laden cells. We further demonstrate that signal transducer and activator of transcription 5 (Stat5), a PRL signal transducer, activates aP2 promoter in a PRL dependent manner. These results suggest that PRL acts as an adipogenesis enhancing hormone in NIH-3T3 cells. PMID- 10674403 TI - Calmodulin antagonists inhibit insulin-stimulated GLUT4 (glucose transporter 4) translocation by preventing the formation of phosphatidylinositol 3,4,5 trisphosphate in 3T3L1 adipocytes. AB - It has been previously reported that calmodulin plays a regulatory role in the insulin stimulation of glucose transport. To examine the basis for this observation, we examined the effect of a panel of calmodulin antagonists that demonstrated a specific inhibition of insulin-stimulated glucose transporter 4 (GLUT4) but not insulin- or platelet-derived growth factor (PDGF)-stimulated GLUT1 translocation in 3T3L1 adipocytes. These treatments had no effect on insulin receptor autophosphorylation or tyrosine phosphorylation of insulin receptor substrate 1 (IRS1). Furthermore, IRS1 or phosphotyrosine antibody immunoprecipitation of phosphatidylinositol (PI) 3-kinase activity was not affected. Despite the marked insulin and PDGF stimulation of PI 3-kinase activity, there was a near complete inhibition of protein kinase B activation. Using a fusion protein of the Grp1 pleckstrin homology (PH) domain with the enhanced green fluorescent protein, we found that the calmodulin antagonists prevented the insulin stimulation of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3] formation in vivo. Similarly, although PDGF stimulation increased PI 3-kinase activity in in vitro immunoprecipitation assays, there was also no significant formation of PI(3,4,5)P3 in vivo. These data demonstrate that calmodulin antagonists prevent insulin-stimulated GLUT4 translocation by inhibiting the in vivo production of PI(3,4,5)P3 without directly affecting IRS1- or phosphotyrosine-associated PI 3-kinase activity. This phenomenon is similar to that observed for the PDGF stimulation of 3T3L1 adipocytes. PMID- 10674405 TI - Inhibitory effect of hexapeptide (RGRHGD) on platelet aggregation. AB - The B chain of beta-bungarotoxin 1-6 sequence, RGRHGD, presents the highest local average hydrophilicity measured by Kyte and Doolittle modeling analysis. The RGRHGD holds parts of both RGD and KGD peptides, which have been reported as having high binding affinity to GPIIb-IIIa. The present study evaluates whether the synthesized hexapeptide, RGRHGD, has an antiplatelet effect and further elucidates the possible mechanisms of action. RGRHGD dose-dependently inhibited rabbit platelet aggregation and adenosine triphosphate release induced by arachidonic acid, collagen, platelet-activating factor, thrombin, or U46619 with the IC50 range of 82.7 to 510 microg/mL. The platelet thromboxane B2 formation induced by collagen or thrombin was also significantly decreased by RGRHGD, but there was no effect on arachidonic acid-induced thromboxane B2 formation. In addition, RGRHGD also inhibited the rise of intracellular calcium level stimulated by arachidonic acid, collagen, or thrombin in Fura 2-AM-loaded platelets. The adenosine 3',5'-cyclic monophosphate level of washed platelets was not affected by RGRHGD. In conclusion, these data indicate that the inhibitory effect of RGRHGD on platelet aggregation may be due to the attenuation of thromboxane A2 formation and intracellular calcium mobilization. In addition, this study may provide a useful method of finding potential therapeutic agents by using molecular modeling analysis. PMID- 10674404 TI - Suppression of transcription factor Egr-1 by curcumin. AB - The transcription factor early growth response-1 gene product (Egr-1) is a member of the family of immediate early response genes and regulates a number of pathophysiologically relevant genes in vasculature that are involved in growth, differentiation, immune response, wound healing, and blood clotting. In the present study, we investigated the effect of curcumin, a natural plant phenolic compound known to exhibit anticarcinogenic, antioxidant, and antiinflammatory properties, on Egr-1 expression in endothelial cells and fibroblasts. Gel mobility shift assays showed that pretreatment of endothelial cells and fibroblasts with curcumin suppressed phorbol 12-myristate 13-acetate and serum induced Egr-1 binding activity to the consensus Egr-1 binding site and also to the Egr-1 binding site present in the promoter of tissue factor gene. Western blot analysis revealed that curcumin inhibited phorbol 12-myristate 13-acetate induced de novo synthesis of Egr-1 protein in endothelial cells. Suppression of Egr-1 protein expression in curcumin-treated cells stemmed from the suppression of Egr-1 mRNA. Northern blot analysis showed that curcumin inhibited serum and phorbol 12-myristate 13-acetate induced expression of tissue factor and urokinase type plasminogen activator receptor mRNA in fibroblasts. Cumulatively, the data show that curcumin suppresses the induction of transcription factor Egr-1 and thereby modulates the expression of Egr-1-regulated genes in endothelial cells and fibroblasts. PMID- 10674406 TI - Differential in vitro effects of the platelet glycoprotein IIb/IIIa inhibitors abixicimab or SR121566A on platelet aggregation, fibrinogen binding and platelet secretory parameters. AB - The aim of this study was to compare fibrinogen binding, inhibition of platelet aggregation and secretory potential of the MAb abciximab (0.5-5 microg/mL) and the peptidomimetic compound SR121566A (15-250 ng/mL) in vitro in whole blood. Fibrinogen binding was followed by flow cytometry; platelet function was evaluated by light transmittance and by impedance aggregometry. Secretory functions of platelets were evaluated using ATP as marker for early secretion by dense granulae and P-selectin (CD62) for alpha-granular secretion as well as CD63 for lysosomal degranulation. Results showed that fibrinogen binding induced by 5 microM TRAP was maximally inhibited greater than 80% at 3 microg/mL abciximab or at 250 ng/mL SR121566A. At these concentrations of antagonists, platelet aggregation induced by 5 microM ADP or 2 microg/mL collagen was inhibited completely. Expression of CD62 was reduced 34% with abciximab or 15% with SR121566A; CD63 expression was reduced 22% with both agents. With both agents, the EC50 for inhibition of CD62 and CD63 expressions was in similar magnitudes than the EC50 for fibrinogen binding inhibition. With 3 microg/mL abciximab, ATP secretion was maximally reduced to 50% of the control, whereas SR121566A at 250 ng/mL had no inhibitory effect on this parameter. A slight increase in ATP secretion was seen with 0.5 microg/mL abciximab and with SR121566A in concentrations of less than 45 ng/mL. The data suggest a discoupling between the anti-aggregatory and the antisecretory effects of IIb/IIIa antagonists. Because it is not established to what extend CD62 or CD63 expression can be reduced by any means, the reduction by 20-30% obtained by 3 microg/mL abciximab or 250 ng/mL SR121566A might already be the maximum possible inhibition by these agents. PMID- 10674407 TI - Relationship between endothelial cell markers and arterial stenosis in peripheral and carotid artery disease. AB - Damage to the endothelium is an important component of atherosclerosis and can be quantified by measuring plasma markers, such as von Willebrand factor, thrombomodulin, intercellular adhesion molecule-1, and E-selectin. We hypothesized that increased levels of these markers would be related to objectively defined disease severity among patients with peripheral atherosclerosis or carotid atherosclerosis. To test this, we measured the markers by using ELISA in the plasma of 45 patients with intermittent claudication alone and in 53 patients presenting with transient ischemic attack. Disease severity in the former was by ankle-brachial pressure index and in the latter by ultrasound defined % stenosis. Any symptomatic dual disease or history or present coronary atherosclerosis warranted exclusion. Data were correlated according to Spearman's method. The only significant correlation was between von Willebrand factor and ankle-brachial pressure index (r = -0.39, p = 0.008). Our data suggest that von Willebrand factor is the most sensitive marker of peripheral atherosclerosis and that none of the plasma markers seems to be a useful marker of the degree of carotid artery stenosis. PMID- 10674408 TI - Antiplatelet effects of sodium nitroprusside in flowing human blood: studies under normoxic and hypoxic conditions. AB - We explored the ability of sodium nitroprusside to modify adhesive and cohesive function of platelets in flowing blood, under normoxic and hypoxic conditions. Aliquots of both untreated and sodium nitroprusside-treated blood were prepared for studies of: (1) platelet aggregation in plasma; (2) erythrocyte deformability; (3) platelet interaction with damaged subendothelium, by using a well-defined perfusion system; and (4) blood gasometry in the perfused samples. Results showed that sodium nitroprusside-treated blood always showed a totally inhibited arachidonic acid-induced platelet aggregation in plasma, as well as significantly increased erythrocyte deformability (0.44+/-0.09 up to 0.66+/-0.05; p<0.05). However, treatment with sodium nitroprusside did not modify the pattern of platelet interaction with subendothelium (percentage of contact, adhesion, thrombus, and covered surface) with respect to untreated blood, under any of the shear rates used (300, 800, and 1800 seconds(-1)), although it significantly reduced the height of thrombi (9.8+/-0.4 vs. 8.3+/-0.4 microm; p<0.05). Hypoxic conditions did not have a noticeable effect in modifying antiplatelet effects of sodium nitroprusside. Additionally, the presence of sodium nitroprusside impaired the normal oxygenation of the blood during perfusion. pO2 in control untreated samples rose from 40.3+/-5.0 mm Hg perfusions to 100.4+/-12.5 mm Hg but remained at 66.3+/-6.3 mm Hg in sodium nitroprusside-treated blood (p<0.05). Our results did not show a significant effect of sodium nitroprusside in the modulation of platelet interaction with subendothelium. The marginal reduction in the thrombi height could be related to rheological interference of increased erythrocyte deformability. PMID- 10674409 TI - Effect of plasminogen activator inhibitor-1 4G/5G polymorphism in Turkish deep vein thrombotic patients with and without FV1691 G-A. AB - A decreased fibrinolytic activity due to increased levels of plasminogen activator inhibitor-1 has been shown in deep vein thrombosis patients. Elevated plasma plasminogen activator inhibitor-1 levels are associated with the 4G allele of a 4G/5G polymorphism located in the promoter region of the plasminogen activator inhibitor-1 gene. Because there is no existing data in the Turkish population, we aimed to study these mutations in patients with deep vein thrombosis (n = 136) and normal controls (n = 113), consecutively selected among unrelated healthy subjects without personal and familial history of atherothrombosis from Ankara, Turkey. DNA was extracted by conventional methods, and polymerase chain reaction of the plasminogen activator inhibitor-1 4G/5G polymorphism was performed according to a previously described method. Genotype distributions of FV 1691G-A and plasminogen activator inhibitor-1 4G/5G are as follows: plasminogen activator inhibitor-1 4G (patients) 0.562, plasminogen activator inhibitor-1 4G (controls) 0.50 (p = 0.6); FV1691A (patients) 0.147, FV1691A (controls) 0.035 (p = 0.005). Our data indicated that plasminogen activator inhibitor-1 4G/5G does not have an effect on the thrombotic risk. Carrying the 4G allele either in heterozygous or homozygous state increases the risk in the presence of FV1691A (odds ratio: 9.8 and 6.9, confidence interval 95% 2.9-32.7 and 1.3-35.8). FV1691A is an independent risk factor for thrombosis (odds ratio: 5.5, confidence interval: 95% 2.5-12.1). We concluded that coexistence of FV1691A and plasminogen activator inhibitor-1 4G allele leads to an increased risk for thrombosis leading a further evidence to another prothrombotic factor that may be necessary for the development of a manifest thrombotic event. PMID- 10674410 TI - A simple screening assay for certain fibrinolysis parameters (FIPA). AB - Hemostasis, the system of generation and degradation of thrombi, consists of coagulation and fibrinolysis. Whereas global assays to study coagulation have existed for many years, there has been no simple, rapid, and economic routine test for the plasmatic fibrinolysis parameters plasminogen activator inhibitor-1, alpha2-antiplasmin, plasminogen, and aprotinin. Here a fast functional global assay for these plasmatic fibrinolytic parameters is presented. However, the present assay is not sensitive to physiological concentrations of prourokinase or tissue-type plasminogen activator. The following assay conditions have been found to be optimal: 50 microL of citrated plasma is incubated with 50 microL of 10 IU urinary-type plasminogen activator (urokinase)/mL, 1.1 mmol/L tranexamic acid, 1% polygelin, 0.1% Triton X-100, phosphate-buffered saline, pH 7.4, for 20 min at 37 degrees C (plasmin generation phase). Then 50 microL of 3 mmol/L HD-Nva-CHA-Lys pNA, 1.05 mol/L KCl is added, and deltaA (405 nm)/10 min (37 degrees C) is determined, by using a microtiterplate reader (plasmin detection phase). The results are calibrated against pooled normal plasma (100% plasmatic fibrinolytic parameters activity). The intra- and interassay coefficients of variation have been found to be less than 5%. The detection limit (sensitivity) of the functional fibrinolysis assay is 5 % of the normal plasmatic fibrinolysis parameters activity. The normal plasmatic fibrinolysis parameters activity is 100%, sigma = 25%. The plasmatic fibrinolysis parameters activity correlates negatively (r = -0.684) with the plasminogen activator inhibitor-1 activity of patient samples. The plasmatic fibrinolysis parameters assay is a simple, rapid, and economic functional test for several clinical relevant fibrinolysis parameters. PMID- 10674411 TI - Antiplatelet drugs attenuate progression of carotid intima-media thickness in subjects with type 2 diabetes. AB - The intima-media thickness of the carotid artery has been established as a surrogate of definite atherosclerosis in subjects with high risk of vascular events. This study was done to evaluate the effectiveness of long-term antiplatelet therapy in attenuating progression of the intima-media thickness of the carotid artery of subjects with type 2 diabetes. Subjects who had an intima media thickness over the threshold of the normal subjects but showed no symptoms of vascular events were randomly divided into groups given antiplatelet drugs [ticlopidine (n = 34) or a small dose of aspirin (n = 40)] or no drugs (n = 74). For the follow-up period (3.0+/-0.06 years), the subjects not given antiplatelet drugs showed a significantly higher progression of intima-media thickness (0.067+/-0.009 mm/year) than those given ticlopidine (0.034+/-0.013 mm/year) or aspirin (0.033+/-0.010 mm/year). Stepwise multivariant regression analysis showed that long-term administration of ticlopidine or aspirin significantly reduced the progression of intima-media thickness of diabetic subjects by 0.041 mm/year or 0.032 mm/ year, respectively. These data indicated that despite differences of their pharmacological mechanisms, antiplatelet drugs could attenuate the progression of intima-media thickness of the carotid artery wall of asymptomatic type 2 diabetics who had early-stage carotid atherosclerosis. PMID- 10674412 TI - Ethanol downregulates transcription of the PAI-1 gene in cultured human endothelial cells. AB - Human endothelial cells are a major site of synthesis for plasminogen activator inhibitor type-1. Elevated plasminogen activator inhibitor type-1 levels in young survivors of myocardial infarction [1] suggest that plasminogen activator inhibitor type-1 may have an important pathologic role in the development of coronary artery disease. Epidemiological studies indicate that moderate alcohol consumption (1-2 drinks/day) reduces the risk for cardiovascular mortality. This cardioprotective benefit has been attributed in part to an increase in fibrinolysis, which decreases fibrin-based thrombosis. The studies described herein were performed to determine whether moderate levels of ethanol affect plasminogen activator inhibitor type-1 gene expression. Cultured human endothelial cells were exposed to 0.1% v/v ethanol for 1 hour. Following incubation in the absence of ethanol plasminogen activator inhibitor type-1, mRNA levels were decreased in a time- and dose-dependent manner, reaching a maximum decrease of 3- to 4-fold at 2 to 4 hours following ethanol challenge. This decline in mRNA occurs at the transcription level; therefore, nuclear transcription run-on assays were performed. A 2.5- to 5-fold decrease in the rate of plasminogen activator inhibitor type-1 gene transcription was measured at 2 and 4 hours following ethanol challenge. Next, a 3.4- and a 1.1-kb fragment from the plasminogen activator inhibitor type-1 promoter region were linked to a luciferase reporter gene, and these constructs were transfected into human endothelial cells. Treatment of these transiently transfected human endothelial cells with ethanol showed a 2- to 3.5-fold decrease in promoter activity, respectively. These results indicate that low doses of ethanol downregulate transcription of the plasminogen activator inhibitor type-1 gene in cultured human endothelial cells. However, the mechanism(s) for this transcriptional decrease is currently unknown. PMID- 10674413 TI - Effects of argatroban on thrombin-induced events in cultured vascular smooth muscle cells. PMID- 10674414 TI - Dermatan sulphate, heparin cofactor II, and F1+2 peptide in non-insulin-dependent diabetes mellitus. PMID- 10674415 TI - Information and communication systems for the assistance of carers based on ACTION. AB - Recent advances in telecommunication technologies allow the design of information and communication systems for people who are caring for others in the home as family members or as professionals in the health or community centres. The present paper analyses and classifies the information flow and maps it to an information life cycle, which governs the design of the deployed hardware, software and the data-structure. This is based on the initial findings of ACTION (assisting carers using telematics interventions to meet older persons' needs) a European Union funded project. The proposed information architecture discusses different designs such as centralized or decentralized Web and Client server solutions. A user interface is developed reflecting the special requirements of the targeted user group, which influences the functionality and design of the software, data architecture and the integrated communication system using video conferencing. ACTION has engineered a system using plain Web technology based on HTML, extended with JavaScript and ActiveX and a software switch enabling the integration of different types of videoconferencing and other applications providing manufacturer independence. PMID- 10674416 TI - Neural network detection of files of suicidal patients and suicidal profiles. AB - The optimal configuration of backpropagation (BP) neural networks was determined after 35 trials with different BP configurations evaluating the total detection rate. Ten different training and testing sets were used to identify optimal samples. All trials included sample files of patients with medically serious suicidal attempts (MSSA) and those of non-suicidal patients. Fifty files were used in each group for training and 49 files for testing with no overlap between the samples. The target variable for training was seriousness of suicide attempt (0 = non-suicidal, 1 = MSSA). The input set included 44 demographic, clinical and patient-history variables. The optimal results showed that 93.8% of MSSA and 89.8% of the non-suicidal patient files were detected. Total success rate (TSR) was 91.8% and positive and negative prediction values (PPV, NPV) were 92% and 95.6%, respectively. Living alone (6.76), treatment compliance (5.86), drug abuse or dependence (2.8), global assessment of functioning (GAF) score (1.49), non paranoid delusions (1.22) and suicide of first degree relative (1.1) were highly associated with MSSA according to the Garson calculation. However, logistic regression attributed high importance to hallucinations (p < 0.0001), diagnosis (p < 0.002), number of children (p < 0.006), GAF score (p < 0.006), employment status (p < 0.02) and stressors (p < 0.03). It was shown that: backpropagation neural networks are very successful in identifying records of MSSA patients; a high GAF score is associated with high risk of MSSA and is the only common variable identified by both methods; and backpropagation identified two non specific factors (living alone and treatment compliance) whereas statistics found specific factors (hallucinations and diagnosis) highly associated with MSSA. PMID- 10674417 TI - The DELPHI method as a consensus and knowledge acquisition tool for the evaluation of the DIABETES system for insulin administration. AB - DIABETES is a decision-support system in the field of insulin administration. System performance evaluation is particularly difficult because of the absence of a uniform decision-making model followed by the specialists. The DELPHI method has been selected since it is appropriate for those domains where there is divergence among experts' opinions. The DELPHI approach helps a number of diabetologists arrive at a consensus and thus it facilitates performance evaluation and further knowledge acquisition. Insulin administration regimes, for 100 diabetic subjects, were proposed by DIABETES and five diabetologists (round 1). These suggestions were compiled and forwarded back to the specialists who proposed a second management approach (round 2). In each case, the experts were asked to justify their decision and comment on the suggestions of their colleagues and DIABETES. A novel scoring system for quantification of agreement was adopted. The DELPHI procedure significantly increased the agreement among the diabetologists from 67% to 84% (X2, p = 0.0001). The agreement between experts' and DIABETES recommendations was to a level of 54%. A total of 3500 comments were acquired by the experts. PMID- 10674418 TI - Intranet-based multi-purpose medical records in orthopaedics. AB - Quality assurance in orthopaedics--as in any medical speciality--relies on precise medical records. Data quality is crucial for statistical evaluation; missing values cannot be avoided but must be minimized. The quality assurance system must be accessible from many locations within the clinic; given the complex and heterogeneous computing infrastructure this is a technological challenge. Intranet technology--the application of internet-tools in local networks--can help to solve the technical problems. A generic Intranet-based quality assurance system in orthopaedics was designed, implemented and evaluated. The basic concept is an intranet data entry form which is generated semi automatically from the data definition. This form is adapted according to the individual needs of the doctors (intelligent data entry). By flexible data transformation the same data set is used for clinical reports as well as scientific evaluations. The first use was for ultrasound examinations of neonatal hips. A report form consisting of 56 items was designed. Within the first 9-month period 1303 cases have been documented. PMID- 10674419 TI - Decision tree induction in the diagnosis of otoneurological diseases. AB - Expert systems have been applied in medicine as diagnostic aids and education tools. The construction of a knowledge base for an expert system may be a difficult task; to automate this task several machine learning methods have been developed. These methods can be also used in the refinement of knowledge bases for removing inconsistencies and redundancies, and for simplifying decision rules. In this study, decision tree induction was employed to acquire diagnostic knowledge for otoneurological diseases and to extract relevant parameters from the database of an otoneurological expert system ONE. The records of patients with benign positional vertigo, Meniere's disease, sudden deafness, traumatic vertigo, vestibular neuritis and vestibular schwannoma were retrieved from the database of ONE, and for each disease, decision trees were constructed. The study shows that decision tree induction is a useful technique for acquiring diagnostic knowledge for otoneurological diseases and for extracting relevant parameters from a large set of parameters. PMID- 10674420 TI - A tool for designing digital test objects for module performance evaluation in medical digital imaging. AB - Currently, medical digital imaging systems are characterized by the introduction of additional modules such as digital display, image compression and image processing, as well as film printing and digitization. These additional modules require performance evaluation to ensure high image quality. A tool for designing computer-generated test objects applicable to performance evaluation of these modules is presented. The test objects can be directly used as digital images in the case of film printing, display, compression and image processing, or indirectly as images on film in the case of digitization. The performance evaluation approach is quality control protocol based. Digital test object design is user-driven according to specifications related to the requirements of the modules being tested. The available quality control parameters include input/output response curve, high contrast resolution, low contrast discrimination, noise, geometric distortion and field uniformity. The tool has been designed and implemented according to an object oriented approach in Visual C++ 5.0, and its user interface is based on the Microsoft Foundation Class Library version 4.2, which provides interface items such as windows, dialog boxes, lists, buttons, etc. The compatibility with DICOM 3.0 part 10 image formats specifications allows the integration of the tool in the existing software framework for medical digital imaging systems. The capability of the tool is demonstrated by direct use of the test objects in case of image processing, and indirect use of the test objects in case of film digitization. PMID- 10674421 TI - Lotka's law and the pattern of scientific productivity in the dental science literature. AB - Statistical regularities can be observed in many natural and social phenomena. From empirical data on the authorship of scientific papers, Lotka deduced an inverse-square law: the number of authors publishing n papers is 1/n2 of those publishing one paper. The general type for the relation (1/n(c)) has a wide range of applicability to a variety of phenomena. This study examined, by means of bibliometric tools, whether Lotka's law could be applied to the literature of dental science. Data came from 20 leading dental science journals, as reported in Journal Citation Reports. The search was performed with a programme developed using Visual Basic for Applications, which counted the number of authors and analysed their contributions to the literature. Authorship for all contributions, as reported in Medline, was compiled for each of these 20 journals for the last 25 years, 1971 through 1995. The total number of authors was 43,796, responsible for 124,556 authorships. The journals published in countries other than the USA exhibited higher degrees of author concentration. The dental science literature conformed very well to Lotka's model with c = 1.95. PMID- 10674422 TI - Memory and mental status correlates of modified Braak staging. AB - We assessed the relationships of performance on memory and mental status tests and neuropathologic stage of Alzheimer's disease as defined by Braak and Braak in 29 patients from a prospective clinicopathologic series. We predicted that memory changes would occur at an earlier Braak stage than mental status changes. Staging was accomplished by matching the topographic distribution of neurofibrillary lesions detected with tau immunocytochemistry to the best fitting diagram published by Braak and Braak. Higher Braak stages were associated with decrements in performance on both memory and mental status tests. As predicted, memory performance declined from stages II to III and mental status did not decline until stages III to IV. The association between memory and Braak stage was unchanged after adjusting for neocortical senile plaques, whereas adjustments for Braak stage eliminated the association between cognitive functioning and amyloid burden. We conclude that Braak staging provides a useful summary of Alzheimer's disease neuropathology, which is associated with both memory and mental status performance. PMID- 10674424 TI - Sex differences in prefrontal volume with aging and Alzheimer's disease. AB - We used volumetric magnetic resonance imaging to examine sex differences in prefrontal tissue volumes of healthy aged and patients with Alzheimer's disease (AD). Healthy subjects had greater total prefrontal volume than AD, and men had greater total prefrontal volume than women (ps < or = 0.02). This was true for both gray and white matter volumes. There were no interactions between group and sex for total prefrontal volume. An exploratory analysis of each group suggested that sex differences in both gray and white matter in healthy aging are not sustained in AD. PMID- 10674423 TI - Evidence for glial-mediated inflammation in aged APP(SW) transgenic mice. AB - Chronic expression of inflammatory cytokines, including interleukin-1beta, tumor necrosis factor alpha, and interleukin-6, by glia may underlie the neurodegenerative events that occur within the brains of patients with Alzheimer's disease (AD). The present study determined whether these markers of inflammation could be observed within the brains of Tg(HuAPP695.K670N/M671L)2576 transgenic mice (Tg2576) that have recently been shown to mimic many features of AD. Interleukin-1beta- and tumor necrosis factor alpha-immunopositive microglia were localized with thioflavine-positive (fibrillar) Abeta deposits. Moreover, interleukin-6 immunoreactive astrocytes surrounded fibrillar Abeta deposits. These findings provide evidence that Tg2576 mice exhibit features of the inflammatory pathology seen in AD and suggest that these mice are a useful animal model for studying the role inflammation may play in this disease. PMID- 10674425 TI - L-type calcium channels in the hippocampus and cerebellum of Alzheimer's disease brain tissue. AB - There is growing evidence that the selective neuronal cell death observed in Alzheimer's Disease (AD) is the result of dysregulation of intracellular calcium (Ca2+) homeostasis. In the present study, L-type voltage sensitive calcium channels (L-VSCCs) were examined in the cerebellum and hippocampus of AD (n = 6; postmortem interval less than 5 h) and age-matched control (n = 6) tissue by homogenate binding techniques and quantitative in vitro receptor autoradiography using [3H]isradipine (PN200-110). Saturation analyses of the cerebellum revealed unaltered [3H]isradipine binding parameters (Kd and Bmax) between AD and control subjects. Analysis of AD and control hippocampus demonstrated significant differences as [3H]isradipine binding increased (62%) in AD, whereas hippocampal cell density decreased (29%) in AD, relative to control subjects. Moreover, AD differentially affected L-VSCC in area CA1 and dentate gyrus. The dentate gyrus had greatly increased binding (77%) with little cell loss (16%) in AD brains, whereas area CA1 had increased binding (40%) with significant cell loss (42%) in AD brains, relative to controls. The results of the present study suggest that hippocampal area CA1 may experience greater cell loss in response to increased L VSCCs in AD relative to other brain regions. PMID- 10674426 TI - Decline in visual attention and spatial memory in aged rats. AB - The present study was a longitudinal study of age-related changes in performance of the 5-choice serial reaction time task, a test of visual attention. Following acquisition of the task, animals were tested on two occasions on their ability to perform the 5-choice task. In Test 1 (Young: 7 months; Aged: 13-14 months) no age related effects on baseline performance were revealed. However, increasing the attentional load of the task revealed an impairment in choice accuracy by animals of the Aged group. In Test 2 (Young: 10-11 months; Aged 23-24 months), animals of the Aged group were significantly impaired on the baseline schedule of the task compared to the Young group. The deficit in accuracy on the task could be improved in the Aged animals by decreasing the attentional load. The results of the present study suggest a deficit in attentional function as a result of the aging process, markedly similar to that observed following lesions of the basalo cortical cholinergic system. PMID- 10674427 TI - Impairments in acquisition and reversals of two-choice discriminations by aged rhesus monkeys. AB - The ability to learn and perform reversals of two object, two patterns, and one spatial discrimination was examined in eight aged (28-34 years), and four adult (8-13 years) behaviorally naive monkeys. As a group, the aged monkeys demonstrated significant difficulties in learning and reversing some of the visual discrimination problems, but had no difficulty learning or reversing the spatial discrimination. Additional analyses revealed that an impairment in learning an object discrimination by the aged monkeys was characterized by a prolonged period of chance performance, and the impairments in performing visual discrimination reversals was related to difficulties in two distinct stages of reversal learning. Despite age-related differences, there was considerable variability in performance among the aged monkeys. These experiments provide the first evidence of significant impairments in learning and reversing visual discriminations by aged monkeys that have not had prior exposure to complex behavioral tasks. PMID- 10674428 TI - Age-related working memory deficits in the allocentric place discrimination task: possible involvement in cholinergic dysfunction. AB - It is well known that learning and memory ability declines with aging. Age related long-term changes in learning and memory ability in rats were investigated with the place navigation task and the allocentric place discrimination task (APDT) in a water maze using the same animals for each task. In a working memory place navigation task, aged animals could learn the location of the platform as well as when they were young, although strategy shifts were observed. In contrast, accuracy in the APDT significantly declined from 90% to 65% with aging. This impairment was ameliorated by an acetylcholine esterase inhibitor physostigmine at 22-23 months old. No amelioration was, however, detected in the same animals tested when they further aged to 26-27 months old. These results suggest that the APDT performance is sensitive to age-related memory deficits and that this may be due to the cholinergic dysfunction. PMID- 10674429 TI - Effects of advanced aging on plasma catecholamine responses to the cold pressor test. AB - Increased basal norepinephrine (NE) concentrations have been demonstrated repeatedly in human aging, but these studies have included almost exclusively "early aging" subjects younger than age 75. We asked if "advanced aging" (over age 80) enhanced the effects of early aging on plasma NE and epinephrine (EPI) concentrations at rest and in response to the cold pressor test (CPT). Eight medically well, cognitively intact advanced aging subjects (84.4+/-0.9 years), 28 medically well cognitively intact early aging subjects (70.3+/-1.3 years), and 19 medically well young subjects (25.4+/-0.9 years) were studied. Both basal NE and the acute NE increase after CPT were significantly higher in advanced aging than in either early aging or young subjects. Plasma EPI concentrations were higher in the advanced aging group than in the other groups and an acute plasma EPI increase after CPT occurred only in the advanced aging group. These results suggest specific effects of advanced aging on both the sympathoneural and sympathoadrenomedullary components of the sympathetic nervous system. PMID- 10674430 TI - Age-related changes in oxidative mechanisms and LTP are reversed by dietary manipulation. AB - Aged rats exhibit an impaired ability to sustain long-term potentiation in dentate gyrus which correlates with a decrease in arachidonic acid concentration. Here we confirm the previous finding that dietary supplementation with arachidonic acid and its precursor, gamma-linolenic acid, reversed the impairment in LTP in aged rats and report that there is a significant correlation between membrane arachidonic acid concentration and response to tetanic stimulation. We observed that age was associated with decreases in the concentration of vitamins C and E and increased activity of superoxide dismutase, indicative of a compromise in antioxidative defenses; these changes were paralleled by increases in interleukin-1beta (IL-1beta) concentration and lipid peroxidation. Dietary manipulation restored polyunsaturated fatty acid concentrations to values observed in tissue prepared from young rats and reversed the age-related changes in vitamins E and C, IL-1beta concentration and superoxide dismutase activity. We propose that these changes reverse the increase in lipid peroxidation and thereby the age-related change in polyunsaturated fatty acids. PMID- 10674431 TI - Age-related changes in LTP and antioxidant defenses are reversed by an alpha lipoic acid-enriched diet. AB - Among the age-related changes identified in rat hippocampus are impairments in LTP and glutamate release. These deficits have been coupled with decreased arachidonic acid concentration. In this study we compared LTP and glutamate release in groups of aged and young rats fed for 8 weeks on a control diet or on a diet enriched in alpha-lipoic acid. Dietary supplementation in aged rats restored hippocampal arachidonic acid concentration to levels observed in tissue prepared from young rats. We observed that aged rats that received the experimental diet sustained LTP in perforant path-granule cell synapses in a manner indistinguishable from young rats whereas the age-related impairment in glutamate release was reversed in synaptosomes prepared from dentate gyrus obtained from these rats. The evidence presented supports the hypothesis that the alpha-lipoic acid-enriched diet has antioxidant properties, because the age related increase in superoxide dismutase activity and decrease in alpha tocopherol concentration were reversed. The finding that the age-related increase in interleukin-1 (IL-1)beta concentration was also reversed suggests a possible role for this cytokine in ageing. PMID- 10674432 TI - Aging does not affect the sleep endocrine response to total sleep deprivation in humans. AB - Aging is associated with decreased sleep continuity, slow wave sleep (SWS), growth hormone (GH) release and an increased hypothalamo-pituitary-adrenocortical (HPA) system activity. Total sleep deprivation (TSD) is a strong stimulus for sleep. To determine if aging affects the response to TSD, for the first time the combined effects of TSD on conventional and spectral sleep electroencephalographic (EEG) parameters and GH, cortisol and prolactin secretion were compared in elderly (60-80 years; n = 7) vs. younger subjects (20-30 years; n = 7). MANOVA revealed a reduction of SWS in the elderly. TSD led to an increase in SWS, a decrease in sleep onset latency, rapid eye movement (REM) density and by trend REM-latency without a global group difference. GH was reduced, whereas prolactin was enhanced in the elderly. After TSD GH was unchanged and prolactin secretion was enhanced without group difference. Thus, the plasticity of the sleep-endocrine system in response to TSD is sustained during aging. The possible involvement of the GABAergic system, that seems not to be severely impaired with age, is proposed. PMID- 10674433 TI - EEG delta power during sleep in young and old rats. AB - Delta EEG power density, which has been viewed as a measure of intensity of NREM sleep, declines across the lifetime in humans, cats, and hamsters, but data in rats have been unclear. It is also uncertain whether older rats differ from younger animals in the degree of change in delta power during recovery sleep following short-term sleep deprivation. We have examined delta power density in NREM sleep under baseline conditions and following 48 h of sleep deprivation in young (3 months), middle-aged (12 months), and older (24 months) rats. The presence or absence of age effects was highly dependent on the method of normalizing the data. When expressed as a fraction of total NREM EEG power, there was no age effect on baseline delta power density, or on the change from baseline to recovery conditions. When expressed as a multiple of delta power in REM under the same condition, the younger rats had higher delta power density than the middle-aged and older rats. For all the ages combined, there was an increase in delta power density in the recovery condition. When examined by age, the younger rats (which started from a higher level of delta power density than the other groups) did not have an increase in delta during recovery; the middle-aged rats tended to, and the older rats (which started from lower baseline levels) significantly increased delta power density in the recovery condition. This suggests that the lower delta power seen during baseline in older rats is not due to decreased ability to generate delta activity. PMID- 10674434 TI - Excitability and branching of neuroendocrine cells during reproductive senescence. AB - In the mollusc Lymnaea stagnalis, neuroendocrine caudodorsal cells (CDCs) were studied physiologically and morphologically from egg layers (EL) (aged 154-400), and animals 4 weeks (CEL-4) (342-455 days), and 8 weeks (CEL-8) (477-660 days) after production of their last egg mass. After recording chemical transmission, electrical coupling and stimulation induced afterdischarges (ADs), CDCs then were filled with Lucifer Yellow. Based on the axonal branching revealed by Lucifer Yellow, CDCs were classified as extensively, moderately, or minimally branched. In EL-CDCs, induction of AD activity, which normally (9) precedes egg-laying, only was initiated in the resting state. CEL-4 CDCs exhibited ADs whereas CEL-8 CDCs did not. CEL-8 CDCs exhibited significantly reduced chemical and electrical transmission, and CEL-4 CDCs did not differ from resting state EL-CDCs. CDC branching was significantly reduced with both increasing age and declining egg laying. Minimally branched CDCs most frequently failed to exhibit an AD and exhibited reduced electrical coupling. We conclude that both physiology and morphology of CDCs are related to age and reproductive state. PMID- 10674435 TI - NGF expression in the aged rat pineal gland does not correlate with loss of sympathetic axonal branches and varicosities. AB - The factors that determine the ability of some, but not all neurons, to sustain their axonal projections during aging remain largely unknown. Because sympathetic neurons remain responsive to nerve growth factor (NGF) in old age, it has been proposed that the selective decrease observed in the sympathetic innervation to some targets in aged rats may be the result of a deficit in target-derived NGF. In this study we utilized two different techniques to demonstrate decreased target innervation by sympathetic fibers in the aged rat pineal gland, which is an appropriate and relevant model for examining mechanisms of neuron-target interactions in aging. Tyrosine hydroxylase immunoreactive profiles were quantified in pineal glands of young and aged male Sprague-Dawley rats. The density of tyrosine hydroxylase-immunoreactive fibers was 30% lower in aged pineals, although the remaining fibers contained 20% more tyrosine hydroxylase immunoreactivity. Othograde tracing of the pineal sympathetic innervation using biotinylated dextran revealed that average axon length, varicosity numbers, branch point numbers, and numbers of terminations were all decreased by approximately 50% in aged tissues, indicating possible functional deficits. These findings suggest that whole branches, along with their associated varicosities were lost in old age. A sensitive quantitative ribonuclease protection assay and a two-site ELISA assay were used to examine whether reduced NGF availability might correlate with sympathetic nerve atrophy. No significant differences were detected in either NGF mRNA or NGF protein levels when comparing young and aged pineal glands, suggesting that atrophy in aged sympathetic neurons is not causally related to reduced availability of NGF at the target. Our results indicate that mechanisms other than NGF expression need to be explored in order to explain the age-related axonal regression observed in this target. PMID- 10674436 TI - Influence of aging on thromboxane A2 and prostacyclin levels in rat hippocampal brain slices. AB - We have investigated the influence of age (3, 18, 24 months) on Thromboxane A2 (TXA2) and Prostacyclin (PGI2) levels in hippocampal slices from F344/NHSD rats. A significant increase in TXA2 and PGI2 levels was observed in 18 and 24 months old compared to 3 months old animals. A significant reduction in the ratio TXA2/PGI2 produced by a higher increase in PGI2 was observed in 24 month old animals. The reduction in the TXA2/PGI2 ratio has been related to vasodilatory and antiaggregating effects that may contribute to protect the brain against neuronal damage. PMID- 10674437 TI - Age differences in the response to the formalin test in rats. AB - We report the results of a study designed to assess age differences in the response to the formalin test, a model of tissue injury and inflammation, while controlling for differences in weight and motoric abilities in three groups of adult male Long-Evans rats: young (3 months old), middle-aged (18 months old), and old (24 months old). The first part of the study assessed initial differences in responsivity and found that the middle-aged group showed the greatest response, whereas the young and old groups did not differ from each other. In the second part of the study, the young and middle-aged animals were followed for a 4 month period. The formalin test was repeated at 2-month intervals. These results indicate that there may be an age-associated change in the sensitivity to tonic pain and that this sensitivity may peak at mid-life. PMID- 10674438 TI - Long-term effects of pergolide and (-)-deprenyl on 3H-mazindol and 3H-spiperone binding in rat brain. AB - The effects of long-term administration of the putative neuroprotective agents pergolide and (-)-deprenyl was assessed by studying 3H-mazindol and 3H-spiperone binding at 12 and 20 months in the major dopamine brain regions. Male Wistar rats were treated from 3 to 20 months, together with their respective untreated and saline injected control groups. The main findings were: 1) there was a decrease in both 3H-mazindol and 3H-spiperone binding with age between 12 and 20 months; 2) there were no differences at 20 months between the pergolide or the (-) deprenyl treated groups and their controls, thus providing no evidence for long term neuroprotection; and 3) there was a marked decrease in 3H-mazindol binding in the injected controls compared with the untreated controls at both 12 and 20 months. This raises the possibility that mild chronic stress may accelerate the aging of the dopamine system. PMID- 10674439 TI - Urinary diversion: the patient's choice. PMID- 10674440 TI - Follow-up is valuable and effective: true, true and unrelated? PMID- 10674441 TI - The type of urinary diversion after radical cystectomy significantly impacts on the patient's quality of life. AB - BACKGROUND: In this study, we used a previously well-validated survey to assess the impact of different forms of urinary diversion on overall quality of life in patients with bladder cancer. METHODS: A total of 92 patients, having three different forms of urinary diversion after radical cystectomy, completed by mail the SF-36, a validated quality-of-life survey. All patients had local/regional disease at the time of cystectomy and are currently without evidence of disease. Completed surveys were then analyzed into physical (PCS) and mental (MCS) component quality-of-life scores per published protocols. Results were then compared with published age-based norms. RESULTS: A total of 38 men who had cystectomy and ileal neobladder had a mean PCS (+/- SD) of 48.4 (7.8) and a mean MCS of 51.0 (7.4); 16 men and women who had cystectomy and Indiana Pouch had a mean PCS of 48.4 (8.9) and a mean MCS of 55.7 (3.8). None of these results is statistically different from published age- and sex-based population norms. Thirty-eight men who had cystectomy and ileal conduit had a mean PCS of 41.4 (8.5) and a mean MCS of 48.2 (10.7). The PCS is not statistically different from the population-based norm; however, the MCS is significantly decreased from the published norm (P = .01). CONCLUSIONS: Patients with ileal conduits have significantly decreased mental health quality of life whereas patients with continent urinary diversions do not. Therefore, when not medically contraindicated, patients should be offered a continent diversion as the diversion of choice after cystectomy. PMID- 10674442 TI - Primary extremity sarcoma: what is the appropriate follow-up? AB - BACKGROUND: Our objective was to evaluate the effectiveness of follow-up tests for detecting first local and distant recurrences in patients with primary extremity soft tissue sarcoma. METHODS: We retrospectively analyzed all adult cases of primary extremity soft tissue sarcoma (n = 174) treated between 1982 and 1992. Patients were observed every 3 months for 2 years, every 4 months the third year, every 6 months the next 2 years, and annually, thereafter. Each visit consisted of taking the patient's history, a physical examination, a complete blood count, a blood chemistry panel, and a chest x-ray. For high-grade tumors, the primary site was imaged annually when clinically appropriate. RESULTS: Of 141 patients who were assessable, 29 patients developed local recurrence and 57 developed distant recurrence. All but one of the local recurrences was detected on the basis of an abnormal physical examination. Of the 29 patients who developed local recurrence, 25 were resected. Distant metastases were detected because of symptoms in 21 cases. Of the 36 asymptomatic lung recurrences, 30 were detected by follow-up chest x-ray. Of the 36 asymptomatic lung recurrences, 24 patients underwent metastasectomy. The positive and negative predictive values of surveillance chest x-ray were 92% and 97%, respectively. Laboratory testing never led to the detection of recurrence. CONCLUSIONS: Close surveillance by clinical assessment and chest x-ray is appropriate for follow-up observation of patients with primary extremity soft tissue sarcoma. PMID- 10674443 TI - Sentinel node biopsy in ductal carcinoma in situ patients. AB - BACKGROUND: Sentinel lymph node (SLN) mapping is an effective and accurate method of evaluating the regional lymph nodes in breast cancer patients. The SLN is the first node that receives lymphatic drainage from the primary tumor. Patients with micrometastatic disease, previously undetected by routine hematoxylin and eosin (H&E) stains, are now being detected with the new technology of SLN biopsy, followed by a more detailed examination of the SLN that includes serial sectioning and cytokeratin immunohistochemical (CK IHC) staining of the nodes. METHODS: At Moffitt Cancer Center, 87 patients with newly diagnosed pure ductal carcinoma in situ (DCIS) lesions were evaluated by using CK IHC staining of the SLN. Patients with any focus of microinvasive disease, detected on diagnostic breast biopsy by routine H&E, were excluded from this study. DCIS patients, with biopsy-proven in situ tumor by routine H&E stains, underwent intraoperative lymphatic mapping, using a combination of vital blue dye and technetium-labeled sulfur colloid. The excised SLNs were examined grossly, by imprint cytology, by standard H&E histology, and by IHC stains for CK. All SLNs that had only CK positive cells were subsequently confirmed malignant by a more detailed histological examination of the nodes. RESULTS: CK IHC staining was performed on 177 SLNs in 87 DCIS breast cancer patients. Five of the 87 DCIS patients (6%) had positive SLNs. Three of these patients were only CK positive and two were both H&E and CK positive. Therefore, routine H&E staining missed microinvasive disease in three of five DCIS patients with positive SLNs. In addition, DCIS patients with occult micrometastatic disease to the SLN underwent a complete axillary lymph node dissection, and the SLNs were the only nodes found to have metastatic disease. Of interest, four of the five node-positive patients had comedo carcinoma associated with the DCIS lesion, and one patient had a large 9.5-cm low grade cribriform and micropapillary type of DCIS. CONCLUSIONS: This study confirms that lymphatic mapping in breast cancer patients with DCIS lesions is a technically feasible and a highly accurate method of staging patients with undetected micrometastatic disease to the regional lymphatic basin. This procedure can be performed with minimal morbidity, because only one or two SLNs, which are at highest risk for containing metastatic disease, are removed. This allows the pathologist to examine the one or two lymph nodes with greater detail by using serial sectioning and CK IHC staining of the SLNs. Because most patients with DCIS lesions detected by routine H&E stains do not have regional lymph node metastases, these patients can safely avoid the complications associated with a complete axillary lymph node dissection and systemic chemotherapy. However, DCIS patients with occult micrometastases of the regional lymphatic basin can be staged with higher accuracy and treated in a more selective fashion. PMID- 10674444 TI - Biopsy method and excision volume do not affect success rate of subsequent sentinel lymph node dissection in breast cancer. AB - INTRODUCTION: Sentinel lymph node dissection (SLND) is becoming a recognized technique for accurately staging patients with breast cancer. Its success in patients with large tumors or prior excisions has been questioned. The purpose of this study was to evaluate the effect of biopsy method, excision volume, interval from biopsy to SLND, tumor size, and tumor location on SLND success rate. METHODS: Consecutive patients who underwent SLND followed by completion axillary lymph node dissection from October 1991 to December 1995 were analyzed. Included were cases performed early in the series before the technique was adequately developed. Excision volume was derived from the product of three dimensions as measured by the pathologist. Two end points were analyzed: sentinel node identification rate and accuracy of SLND in predicting axillary status. Univariate analyses using chi2 or Fisher's exact test for categorical variables and Wilcoxon rank sums for continuous variables were performed. Multivariate analysis was performed using logistic regression. RESULTS: There were 284 SLND procedures performed on 283 patients. Median age was 55 years. The most recent biopsy method used before SLND was stereotactic core biopsy in 41 (14%), fine needle aspiration in 62 (22%), and excision in 181 (64%) procedures. The mean excision volume was 32 ml with a range of 0.3-169 ml. The mean time from biopsy to SLND was 17 days with a range of 0-140 days. The mean tumor size was 2.0 cm (15 Tis [5%], 184 T1 [65%], 72 T2 [25%], and 13 T3 [5%]). Tumors were located in the outer quadrants in 74%, the inner quadrants in 18%, and subareolar region in 8%. The sentinel node was identified in 81%, and 39% had metastases. There were three false-negative cases early in the series. Sensitivity was 97%, and accuracy was 99%. Negative predictive value was 98% in cases in which the sentinel node was identified. On the basis of biopsy method, excisional volume, time from biopsy to SLND, tumor size, and tumor location, there was no statistically significant difference (P>.05) in sentinel node identification rate or accuracy of SLND. CONCLUSIONS: SLND has a high success rate in breast cancer patients regardless of the biopsy method or the excision volume removed before SLND. In addition, the interval from biopsy to SLND, tumor size, and tumor location have no effect on the success rate of SLND, even in this series which included patients operated on before the technique was adequately defined. Patients with breast cancers located in any quadrant and diagnosed either with a needle or excisional biopsy could be evaluated for trials of SLND. PMID- 10674445 TI - Radioguided sentinel node biopsy to avoid axillary dissection in breast cancer. AB - BACKGROUND: Sentinel node (SN) biopsy may predict axillary status in breast cancer. We retrospectively analyzed more than 500 SN cases, to suggest more precise indications for the technique. METHODS: 99mTc-labeled colloid was injected close to the tumor; lymphoscintigraphy was then performed to reveal the SN. The next day, during surgery, the SN was removed by using a gamma probe. Complete axillary dissection followed, except in later cases recruited to a randomized trial. The SN was examined intraoperatively by conventional frozen section, in later cases by sampling the entire node and using immunocytochemistry. RESULTS: In the first series, the SN was identified in 98.7% of cases; in 6.7%, the SN was negative but other axillary nodes were positive; in 32.1%, the SN was negative by intraoperative frozen section but metastatic by definitive histology, prompting introduction of the exhaustive method. In the randomized trial, the SN was identified in all cases so far, the false-negative rate is approximately 6.5%, and in 15 cases, internal mammary chain nodes were biopsied. CONCLUSIONS: SN biopsy can reliably assess axillary status in selected patients. The problems are the SN detection rate, false negatives, and the intraoperative examination, which can miss 30% of SN metastases. Our exhaustive method overcomes the latter problem, but it is time consuming. PMID- 10674446 TI - Is intensive follow-up really able to improve prognosis of patients with local recurrence after curative surgery for rectal cancer? AB - BACKGROUND: Because more than 90% of local recurrences after curative surgery for rectal cancer appear within the first 36 months after surgery, an intensive and strict follow-up program during this period could improve early diagnosis and, thus, prognosis of patients. METHODS: Of the 216 patients who underwent surgery for rectal cancer, 127 entered an intensive follow-up program (median follow-up: 42 months); the clinical outcome of the remaining 89 patients was reconstructed with the help of their general practitioners. RESULTS: Fifty eight (26.8%) of the 216 patients who were treated with curative surgery alone developed a local recurrence; pelvic recurrences were prevalent. Eleven (30.5%) of the 36 patients who had recurrence during follow-up, and 6 of the 22 who had not undergone follow up, had a reoperation with curative intent; the median survival was 19 months vs. 8 months, respectively (P = ns). Four (44.4%) curative reoperations were performed on the 9 asymptomatic patients and in 13 (26.5%) of the 49 cases with symptomatic local recurrences. Median survival was 15 months vs. 14 months, respectively (P = n.s). All patients except one (living after 42 months from reoperation) died within 48 months. CONCLUSIONS: In our study, adherence to a strict follow-up program unfortunately proved to be ineffective for improving long-term survival for patients who underwent reoperation with curative intent. PMID- 10674447 TI - Preoperative combined radiotherapy and chemotherapy for middle and lower rectal cancer: preliminary results. AB - BACKGROUND: Adjuvant treatment for rectal cancer is still controversial. This study reports on overall survival and disease-free survival, toxicity, downstaging, and surgical morbidity in rectal cancer patients who received combined chemoradiation therapy followed by curative surgery. METHODS: Between 1993 and 1998, 51 patients (31 males and 20 females; median age, 60 years; range, 33-73 years) underwent chemoradiation therapy followed by radical surgery for middle and lower rectal adenocarcinoma. Criteria for giving preoperative radiotherapy (total 45 Gy in 25 fractions of 1.8 Gy/day for 5 weeks) and chemotherapy (5-fluorouracil 350 mg/m2/day and leucovorin 10 mg/m2/day, bolus on days 1-5 and 29-33) were an age younger than 75 years; an Eastern Cooperative Oncology Group performance status score of 0 to 2; and clinical preoperative stage II-III. Forty-three low anterior and eight abdominoperineal resections were performed. Median follow-up time was 29 (range, 3-63) months. RESULTS: Although grade 3 to 4 toxicity occurred in 14 cases (27.4%), all patients completed the planned adjuvant therapy. At pathology, a complete response was found in eight (15.7%) cases. Of the remaining 43 cases, 22 were stage I, 12 were stage II, and 9 were stage III. Five-year actuarial disease-free survival and overall survival rates were 86.4% and 85.5%, respectively. Whereas no local recurrences were found, 4 patients had distant metastases. Three patients died (1 of cancer related causes), 45 are alive and disease free, and 3 are alive with disease. CONCLUSIONS: The combined preoperative chemoradiation approach used by us seems to improve the disease-free survival and overall survival of selected patients with rectal cancer. However, a longer follow-up time is required to confirm these preliminary results. PMID- 10674448 TI - Neoadjuvant chemotherapy with P-ELF (cisplatin, etoposide, leucovorin, 5 fluorouracil) followed by radical resection in patients with initially unresectable gastric adenocarcinoma: a phase II study. AB - BACKGROUND: Gastric cancer is the most frequent gastrointestinal cancer in Mexico. Only 33% of cases are resectable. Our aim was to determine the activity and toxicity of the cisplatin, etoposide, leucovorin, and 5-fluorouracil combination in initially unresectable tumors and to determine its ability to permit resection. METHODS: Sixty patients with unresectable gastric adenocarcinoma were treated with cisplatin 80 mg/m2, etoposide 80 mg/m2, leucovorin 25 mg/m2, and 5-fluorouracil 800 mg/m2 by central intravenous catheter for 4 consecutive days. Two courses of this combination were followed by surgical resection. RESULTS: The overall response rate was 36.8% (20 partial responses and one complete response). By using logistic regression analysis, the tumor, node, and metastasis stage (risk ratio, 2.04; 95% confidence interval, 1.03-4.02; P = .039) was identified as the response determinant to chemotherapy. Major toxicity was grade 3 or 4 neutropenia in 67% of patients. Ten resections were performed (17.5%); five were curative and five palliative. Operative morbidity and mortality rates were 40% and 10%, respectively. The median length of survival was 7.46 and 13.3 months for nonresponders and responders, respectively (P = .011). CONCLUSIONS: The cisplatin, etoposide, leucovorin, and 5-fluorouracil combination is active in advanced gastric cancer and the toxicity level is acceptable. This treatment permits a 17.5% resection rate in previously unresectable tumors. A randomized trial of surgery vs. neoadjuvant chemotherapy plus surgery is warranted. PMID- 10674449 TI - Sentinel node biopsies in melanoma patients: a protocol for accurate, efficient, and cost-effective analysis by preselection for immunohistochemistry on the basis of Tyr-PCR. AB - BACKGROUND: Immunohistochemistry (IHC) of serial sectioning is considered the gold standard for detection of melanoma activity in sentinel node (SN) biopsies. However, this is cost and labor intensive. In contrast, tyrosinase reverse transcription-polymerase chain reaction (RT-PCR) is simple and quick, but it is hampered by its extreme sensitivity. This study was performed to test whether a strategy that combines the two methods, using tyrosinase RT-PCR to preselect nodes for IHC, could be accurate and cost effective. METHODS: In 36 patients, SNs were identified by scintigraphy and patent blue uptake. Of each SN, one cross section was analyzed first by hematoxylin and eosin staining. Next, all nodes were examined by serial sectioning and IHC of one-half and tyrosinase RT-PCR of the other. Before comparison, all results were documented in a blinded manner. Material costs and workload estimates were noted per SN. RESULTS: Fifty-five SNs were retrieved from the 36 patients. Hematoxylin and eosin staining of the first cross section revealed tumor positivity in 3 patients (6 SN). Tyrosinase RT-PCR was positive in 11 of the remaining 33 patients (19 of 49 SN). Of these same 11 patients, only 5 were shown to have tumor-positive SNs by using IHC on serial sections (7 SN). All these nodes had been positive for tyrosinase on PCR. For IHC, an average of 40 sections were prepared and examined per SN at a cost of $200(U.S.)/SN. In contrast, routine tyrosinase RT-PCR costs $37(U.S.)/SN, and takes 5% of the time necessary for IHC. A strategy including hematoxylin and eosin staining on the first cross section, followed by tyrosinase RT-PCR on half of each negative (half) node, could preselect nodes to be taken through serial sectioning. In these series, such a strategy would have prevented serial sectioning and IHC of 30 SN from 22 patients. Apart from a considerable gain in efficiency, this would have reduced material costs by a minimum of $6000 (U.S.). This discrepancy would be even higher if work intensity of analysts and pathologists were considered. CONCLUSIONS: In routine analysis of SN biopsies in melanoma patients, tyrosinase RT-PCR can be used effectively to preselect nodes for further IHC of serial sections. This method seems both time and cost effective. PMID- 10674450 TI - Hilar cholangiocarcinoma: a review and commentary. AB - Hilar cholangiocarcinoma is an uncommon cause of malignant biliary obstruction marked by local tumor spread for which surgery offers the only chance of cure. The diagnostic evaluation and surgical management of this disease continues to evolve. Although direct cholangiography and endoscopic biliary procedures have been used extensively to anatomically define the extent of tumor involvement, establish biliary decompression, and obtain histological confirmation of tumor, reliance on these invasive procedures is no longer necessary, and may be detrimental. Current noninvasive imaging technology permits accurate staging of the primary tumor and has improved patient selection for operative intervention without the need for invasive procedures. Overall survival has improved in accordance with an increasingly aggressive surgical approach. The propensity of this tumor for local invasion has led most experienced hepatobiliary centers to perform a partial hepatectomy in 50% to 100% of cases. Three-year survival rates of 35% to 50% can be achieved when negative histological margins are attained at the time of surgery. When resection is not feasible, either operative bilioenteric bypass or percutaneous transhepatic intubation can achieve significant palliation. There is no effective adjuvant therapy for this disease, and unless clear indications of unresectability exist, most patients should be considered for surgical exploration. PMID- 10674451 TI - Tamoxifen for the prevention of breast cancer in the high-risk woman. PMID- 10674452 TI - Time course of nodal enhancement with CT X-ray nanoparticle contrast agents: effect of particle size and chemical structure. AB - RATIONALE AND OBJECTIVES: Levels of CT enhancement in rabbit lymph nodes were followed with time after subcutaneous injection of four iodinated, insoluble nanoparticle contrast agents to provide experimental support for the hypothesis that clearance of these agents is related to the chemical structure of the agent itself. The impact of particle size was also studied. METHODS: Subcutaneous injections (2 x 0.25 mL) were made in the dorsum of rabbit paws with 15% suspensions of four nanoparticle contrast agents. Images were obtained at 4, 10, 24, 48, and 72 hours and 5, 7, and 14 days after injection. Average attenuation (in Hounsfield units [HU]), node volume, and total iodine uptake were estimated from the CT scans for each lymph node at each time point. RESULTS: All the agents provided adequate enhancement of both the popliteal and axillary lymph nodes of the rabbit (ie, > delta100 HU). Lymph node volume appears to be related to the persistence of enhancement, with long-lived agents demonstrating the greatest increase in size. The rate of clearance from the lymph nodes is related to the structure of the agent. CONCLUSIONS: Clearance of insoluble, iodinated nanoparticle contrast agents from lymph nodes can be modulated by changes in the structure of the agent itself. Using the same agent, smaller particles deliver material to the lymph nodes more quickly and clear more quickly. PMID- 10674453 TI - Intravascular ultrasound evaluation of peripheral arterial stent-grafts. AB - RATIONALE AND OBJECTIVES: To evaluate neointimal hyperplasia, plaque distribution, and morphologic features of peripheral arterial stent-grafts with intravascular ultrasound (IVUS). METHODS: Twenty-three patients with stenoses or occlusions of the pelvic or femoral arteries were treated with 31 stent-grafts. Angiography and IVUS of the stented artery were performed 13.9 +/- 9.7 months after stent implantation. Maximum in-stent restenosis was measured by IVUS. Plaque composition and lesion topography were also assessed. RESULTS: The maximum in-stent restenosis was 53.2 +/- 26.5% for the femoral and 14.2 +/- 10.1 for pelvic arterial stent-grafts. Predilection sites of maximum neointimal tissue accumulation were the edges of the femoral stent-grafts. Only small amounts of neointimal hyperplasia were found in the stent-graft edges. No predilection site for maximum in-stent restenosis was found for the pelvic arterial stent-grafts. CONCLUSIONS: Predilection sites of maximum in-stent restenosis were the edges of femoral stent-grafts in contrast to pelvic stent-grafts. Femoral stent-grafts showed significantly higher graded stenoses with IVUS than iliac stent-grafts. The authors' findings at IVUS did not change the treatment plan in these patients treated with stent-grafts. PMID- 10674454 TI - Comparison of harmonic and conventional power Doppler ultrasonography for assessment of slow flow in hyperechoic tissue: experimental study using a Doppler phantom. AB - RATIONALE AND OBJECTIVES: Despite the advantages of depicting slow flow in small vessels, conventional power Doppler ultrasound (US) has a basic limitation, specifically that artifactual power Doppler signals mimic blood flow, especially in hyperechoic tissue. The purpose of this study was to compare harmonic power Doppler US with power Doppler US using a Doppler phantom under various parameter settings, focusing on the assessment of slow flow in the hyperechoic tissue. METHODS: While controlling the flow velocity (5 and 10 cm/s), pulse repetition frequency (500, 700, and 1,000 Hz), wall filter (low and medium), and Doppler gain (90%, 96%, and 100%), the authors performed both harmonic Doppler US and power Doppler US by using a Doppler phantom/flow control system. We measured and compared the relative intensities of the Doppler signals (0-250 scale) in both the vessels and hyperechoic tissue-mimicking materials with the two different imaging modalities. RESULTS: Power Doppler US with any combination of the four parameters evaluated depicted strong flow signals (mean, 213) that were superior to harmonic Doppler US (mean, 61). Relatively strong artifactual signals within the hyperechoic tissue-mimicking materials were noted on all power Doppler US studies (mean, 106) but nearly none on harmonic Doppler US (mean, 3). The contrast-to-noise ratio of harmonic Doppler US was significantly greater than that of power Doppler US. CONCLUSIONS: Harmonic Doppler US is more useful in assessing slow flow in hyperechoic tissue than power Doppler US because it produces fewer artifactual Doppler signals originating from stationary hyperechoic tissues, which can be misjudged as true signals on power Doppler US. PMID- 10674455 TI - Evaluation of breath-hold contrast-enhanced 3D magnetic resonance angiography technique for imaging visceral abdominal arteries and veins. AB - RATIONALE AND OBJECTIVES: To evaluate the diagnostic value of breath-hold contrast-enhanced 3D magnetic resonance angiography (MRA) for assessment of the visceral abdominal arteries and veins in patients with suspected abdominal neoplasms. METHODS: Twenty-one patients underwent MR imaging on a 1.5 T unit using a body phased-array coil. MRA was performed with a 3D-FLASH sequence (TR 3.8 ms, TE 1.3 ms, flip angle 25 degrees, acquisition time 20 seconds), 8 to 12 seconds after an intravenous bolus injection of Gd-DTPA. The acquisition delay between the arterial and the portal venous phase was 12 seconds. The image quality and the degree of vascular involvement were evaluated using coronal source images and maximum intensity projection reconstructions. Diagnosis was confirmed by surgery/histology. RESULTS: Image quality was optimal in more than 85% of the patients (19/21 arterial phase and 17/21 portal venous phase). MRA correctly predicted vascular status in 20 of 21 patients (95%), with complete concordance between MRA results and surgical findings. In one patient with chronic pancreatitis, MRA demonstrated a false-positive finding that could not be confirmed surgically. CONCLUSIONS. Breath-hold contrast-enhanced 3D-MRA is a valuable technique for assessing visceral abdominal arteries and veins. PMID- 10674456 TI - Enhanced detection of blood flow in the normal canine prostate using an ultrasound contrast agent. AB - RATIONALE AND OBJECTIVES: To evaluate the sonographic appearance of normal prostate vascularity in dogs before and after injection of a new ultrasound contrast agent, NC100100. METHODS: Thirty-five intravenous injections of NC100100, in doses ranging from 0.00625 to 0.05 microL microbubbles/kg, were administered to seven anesthetized mongrel male dogs. Transrectal color Doppler imaging and power Doppler imaging were used to perform the assessment. The visibility of the vascular pattern of the prostate was assessed, including dynamics of contrast inflow, blood flow symmetry, and duration times. RESULTS: Before contrast administration, the vascular pattern was poorly visualized in all cases. After contrast injection, the visibility of the vascular architecture improved significantly for both modalities. Independent of the imaging method used, higher doses tended to be more effective than lower doses. Contrast kinetics in the prostate vessels was demonstrated with a mean time from injection of the ultrasound contrast agent to enhancement of the Doppler signals in the subcapsular arteries (+/-1 SD) of 13+/-3 seconds, and the ultrasound contrast agent reached the central periurethral veins 3 to 6 seconds later. A spokelike radial pattern of internal prostatic vessels observed with enhanced ultrasound could also be seen on silicone microfil x-ray images. The Doppler enhancement persisted for a mean time ( +/-1 SD) of 904 seconds (approximately 15 minutes) +/ 225 seconds and tended to increase with increasing dose. CONCLUSIONS: NC100100 significantly improves the detection of blood flow in the normal canine prostate and allows more accurate depiction of the vascular architecture of the prostate. PMID- 10674457 TI - Multipoint rank-order study methodology: observer issues. AB - RATIONALE AND OBJECTIVES: We performed a multipoint rank-order experiment to evaluate variability in observers' sensitivity to small differences in image presentation and to assess observers' performance as a function of the type and number of tasks included. METHODS: Five experienced observers were presented with four sets of chest images that had been compressed at five different levels. Each set contained six images ranging from noncompressed to approximately 60:1 compressed images. Observers were asked to review all images of each case side by side and rank-order the "quality" of each to enable determination of the presence or absence of interstitial disease and/or pneumothoraces. RESULTS: Observers varied significantly in their ability to detect very small differences among the images (P < 0.001). Those who performed well did so regardless of whether they ranked a specific abnormality in a multidisease or a single-disease setting. CONCLUSIONS: Selected observers can reliably detect very small differences among similar images. These readers could be used to confirm or rule out the need for objective observer-performance-type studies. PMID- 10674458 TI - Rheolytic hydrodynamic thrombectomy for percutaneous treatment of acutely occluded infra-aortic native arteries and bypass grafts: midterm follow-up results. AB - RATIONALE AND OBJECTIVES: To evaluate the efficacy of a rheolytic thrombectomy catheter (RTC) for treatment of acutely occluded infra-aortic native arteries and bypass grafts and to determine midterm primary patency, death, and amputation free survival rates. METHODS: From March 1995 to September 1997, 112 patients with occluded arteries or bypass grafts were primarily treated with RTC at two centers. Thrombus removal was evaluated by two angiographers. RESULTS: More than 75% of the thromboembolic material could be removed with RTC alone. Mean activation time of RTC was 280 +/- 163 seconds. Residual mural or organized thrombi (29%) required adjunctive fibrinolytic therapy or aspiration thrombectomy. Remaining stenoses were treated with percutaneous transluminal angioplasty and additional stent implantation. For acute reocclusions, surgical intervention was required. Technical success after the entire procedure was 88.4%. RTC-associated complications included distal embolization, dissection, vessel perforation, and technical failure of RTC. Mean follow-up time was 14.8 months +/- 11.5, rates of primary patency, secondary patency, death, and amputation-free survival were 60%, 84%, 16%, and 75% after 2 years, respectively. CONCLUSIONS: RTC is a rapid and efficient technique for mechanical thrombectomy of acutely thrombosed native leg arteries and bypass grafts. Midterm results are comparable to the results of alternative treatment modalities such as Fogarty balloon thromboembolectomy or local fibrinolysis. PMID- 10674459 TI - Safety and efficacy of Omniscan (gadodiamide injection) at 0.1 mmol/kg for MRI in infants younger than 6 months of age: phase III open multicenter study. AB - RATIONALE AND OBJECTIVES: To demonstrate that gadodiamide injection is a safe and efficient contrast agent for MRI in infants younger than 6 months of age. METHODS: The authors designed a phase III multicenter nonrandomized study using a control group. Gadodiamide injection at a dosage of 0.1 mmol/kg body weight was administered to 39 children; 20 received no contrast. The mean age was 10.6 weeks in the contrast group and 9.3 weeks in the control group. MR examinations, blood (serum creatinine, S-ASAT, S-ALAT, S-bilirubin, alkaline phosphatase) and urine (proteins, blood, others) sampling before sedation and after examination, heart rate (electrocardiography) and oxygen saturation (pulse oximetry) during examination, adverse events, and efficacy parameters were analyzed. RESULTS: In the contrast group, 18 (51.4%) children had 31 abnormal changes in one or more of the safety parameters and vital signs. In the control group there were 16 (80.0%) children with 19 abnormal changes. Gadodiamide injection had no negative influence on the safety parameters. No serious adverse events occurred, and only three clinically relevant adverse events (elevation of S-ALAT and S-ASAT, elevation of bilirubin) in two patients in the contrast group and one event (vomiting) in one patient in the control group were documented. The benefit of the contrast medium was clearly shown for all evaluated parameters. CONCLUSIONS: Gadodiamide injection is safe, well tolerated, and effective in infants younger than 6 months of age. PMID- 10674460 TI - Noise affects auditory and linguistic processing differently: an MEG study. AB - We investigated the influence of noise on brain responses to spoken sentences in MEG. Sixteen subjects had to listen to acoustically presented sentences and judge their syntactic correctness. Sentences were either presented on a silent background or with noise. Noise had differential effects on early auditory and syntactic processes. While noise affected early auditory processes only in the right hemisphere, noise had a general effect on syntactical processes. The evoked responses to syntactic violations compared with correct sentences, namely an early left anterior negativity, were significantly suppressed when noise was present The noise suppression effect, however, was not lateralized. PMID- 10674461 TI - Inhibition of TNF-alpha can attenuate or exacerbate excitotoxic injury in neonatal rat brain. AB - Tumor necrosis factor (TNF)-alpha, a multifunctional pro-inflammatory cytokine, has been implicated in the pathogenesis of acute ischemic brain injury. Recent data also suggest that TNF-alpha is a clinically relevant mediator of neonatal brain injury. We hypothesized that inhibition of TNF-alpha activity would reduce excitotoxic brain injury in neonatal rats. To test this hypothesis, we evaluated the efficacy of a TNF binding protein (bp) in attenuating NMDA-induced injury in 7 day old rats. Intrastriatal co-injection of TNFbp (3.75 microg) with NMDA (10 nmol) reduced striatal injury by 26%; in contrast, intra-hippocampal co-injection of TNFbp (3.75 microg) with NMDA (10 nmol) increased hippocampal damage by 68%. These findings indicate that TNF-alpha may have both beneficial and deleterious effects in the injured neonatal brain. PMID- 10674462 TI - Alexia for Braille following bilateral occipital stroke in an early blind woman. AB - Recent functional imaging and neurophysiologic studies indicate that the occipital cortex may play a role in Braille reading in congenitally and early blind subjects. We report on a woman blind from birth who sustained bilateral occipital damage following an ischemic stroke. Prior to the stroke, the patient was a proficient Braille reader. Following the stroke, she was no longer able to read Braille yet her somatosensory perception appeared otherwise to be unchanged. This case supports the emerging evidence for the recruitment of striate and prestriate cortex for Braille reading in early blind subjects. PMID- 10674463 TI - Hierarchical visual processing is dependent on the oculomotor system. AB - Using functional MRI and eye movement recordings we studied the processing of hierarchical stimuli. In agreement with others, we found a minor left hemispheric dominance during local and right dominance during global processing. When attention was directed locally, well-known oculomotor cortical areas were activated, and saccades were elicited in 41% of the trials. Their latencies were similar to pro-saccades. During global processing virtually no saccades occurred. These results suggest two different operational modes of attention. Attending to local features induces a shift of attention, which simultaneously computes a saccade on any level above the brainstem with a computational burden equal to reflexive saccades. Conversely, attending to global features induces an expansion of the focus of attention, which reinforces fixation. PMID- 10674464 TI - Quinolinic acid inhibits glutamate uptake into synaptic vesicles from rat brain. AB - Quinolinic acid (QA) is an endogenous and potent neurotoxin associated with the neurotoxicity of various common diseases. The uptake of neurotransmitters into synaptic vesicles is an important event involved in the storage and release of neurotransmitters by vesicles. The influence of QA on the uptake of glutamate, GABA and glycine into rat brain synaptic vesicles was investigated. QA (0.3-10 mM) significantly inhibited (>50%) the uptake of glutamate into synaptic vesicles, whereas QA at concentrations up to 10 mM had no significant effect on GABA or glycine uptake. Such results indicate that QA is able to selectively inhibit the vesicular uptake of glutamate, without interfering with the uptake of the inhibitory neurotransmitters GABA and glycine. These findings might be related to the neurotoxic effects of QA in the brain. PMID- 10674465 TI - Melatonin synthesis: arylalkylamine N-acetyltransferases in trout retina and pineal organ are different. AB - Serotonin N-acetyltransferase (AANAT) is the first enzyme in the conversion of serotonin to melatonin. Changes in AANAT activity determine the daily rhythm in melatonin secretion. Two AANAT genes have been identified in the pike, pAANAT-1 and pAANAT-2, expressed in the retina and in the pineal, respectively. The genes preferentially expressed in these tissues encode proteins with distinctly different kinetic characteristics. Like the pike, trout retina primarily expresses the AANAT-1 gene and trout pineal primarily expresses the AANAT-2 gene. Here we show that the kinetic characteristics of AANAT in these tissues differ as in pike. These differences include optimal temperature for activity (pineal: 12 degrees C; retina: 25 degrees C) and relative affinity for indoleethylamines compared to phenylethylamines. In addition, retinal AANAT exhibited substrate inhibition, which was not seen with pineal AANAT. The kinetic differences between AANAT-1 and AANAT-2 appear to be defining characteristics of these gene subfamilies, and are not species specific. PMID- 10674466 TI - Developmental dependency of Meissner corpuscles on trkB but not trkA or trkC. AB - The distribution of S100-immunoreactive (ir) corpuscular endings was examined in the palate of wildtype and knockout mice for trkA, trkB or trkC. In wildtype mice, S100-ir corpuscular endings were abundant at the top of palatal rugae. The endings contained 2-4 parallel arrays of S100-ir neurites. The distribution of S100-ir nerve endings in trkA and trkC knockout mice was similar to that in wildtype mice; S100-ir corpuscular endings were abundant in palates of the mutant mice. In trkB knockout mice, the palate was devoid of corpuscular endings, An immunoelectron microscopic method indicated that S100-ir corpuscular endings were identical to Meissner corpuscles. The normal development of Meissner corpuscles is probably dependent on trkB but not trkA or trkC. PMID- 10674467 TI - Ipsilateral and contralateral transfer of tactile learning. AB - We examined the spatial organization of perceptual learning in a cortex-dependent task. Rats learned a tactile task using four whiskers on one side of the snout, all others being clipped. These trained whiskers were then clipped and prosthetic whiskers were attached. Subsequent performance was found to be determined by the location of the prosthetic whiskers. There was partial transfer of learning to neighbouring whisker positions. In addition, there was partial transfer of learning to whisker positions on the other side of the snout, but only if the prosthetic whiskers were symmetrically opposite the trained whiskers. These findings suggest that neural changes underlying perceptual learning are distributed according to the topographic organization of the sensory cortical map. PMID- 10674468 TI - Subthalamic nucleus microinjections of 5-HT2 receptor antagonists suppress stereotypy in rats. AB - The subthalamic nucleus (STN) is an important mediator of basal ganglia output. We studied the effects of STN microinjections of the serotonin-2 (5-HT2) antagonists clozapine, mesulergine and M100,907 on apomorphine-induced stereotypic activity in the rat. Each compound profoundly decreased the expression of stereotypic behavior, with particularly strong effects to reduce gnawing behavior. Because M100,907 does not have appreciable affinity for dopamine D1 and D2 receptors, and since all three agents are 5-HT2 antagonists, the current data suggest that basal ganglia output related to orofacial movements can be significantly modified by 5-HT2 receptors. The results suggest that antipsychotics with serotonergic properties may have direct actions on the STN that influence their potential to produce orofacial and other motor side effects. PMID- 10674469 TI - Attentional suppression of activity in the human visual cortex. AB - We have used fMRI to examine the nature of the changes that occur in the human visual cortex when an observer attends to a particular location in the visual image. Previous studies have shown that the magnitude of the response to a visual stimulus is increased when the observer attends to the stimulus. We show that, in addition, attention to a particular location results in a widespread suppression of activity levels at all other locations. This suggests that a key mechanism of attentional modulation may be that spontaneous (baseline) levels of neural activity are adjusted in a position-dependent manner across the entire visual field. PMID- 10674470 TI - Serum immunoglobulins in brain tumours and lumbar disc diseases. AB - Changes of serum immunoglobulin (Ig) concentrations may occur in both brain tumours and lumbar disc diseases (LDD). The purpose of this study was to investigate the changes of pre- and post-operative serum Ig levels in brain tumours and LDDs. Serum IgG, IgA and IgM levels were measured in 127 patients with brain tumour, 100 patients with LDD and 20 healthy subjects without neurological disease. Increases in one or more of the pre-operative serum Ig levels were observed in the patients with both brain tumours and LDDs compared with controls. However pre-operative serum IgG level was highly increased in all brain tumour types and LDDs (p<0.001). Serum IgA levels and IgM levels in the post-operative stage were significantly decreased in patients with acoustic neurinoma (p<0.01, p<0.001, respectively). Post-operative serum IgG, IgA and IgM levels were significantly decreased (p<0.001) in patients with meningioma. Post operative serum IgG and IgM levels were significantly decreased (p<0.001) in patients with glioma. Patients with LDD showed a significantly decline in post operative serum IgA and IgM levels (p<0.001). We think that decline in post operative serum Ig levels may be of prognostic value in the patients with brain tumours and LDDs. PMID- 10674471 TI - Denervation and NKI receptor block modulate stimulated CGRP and PGE2 release from rat skin. AB - We investigated the possible neurogenic origin of prostaglandin E2 (PGE2) in the rat skin, in vitro. The hairy skin of one hindpaw was denervated and one week later the dorsal hindpaws were skinned to study the release of calcitonin gene related peptide (CGRP) and PGE2 using the EIA technique. Stimulation with bradykinin (BK) caused a significant release of CGRP (1.4-fold increase) and PGE2 (3-fold) which was massively augmented under neurokinin I (NKI) receptor antagonist treatment (CGRP: 4-fold, PGE2: 5-fold). In denervated skin the BK evoked CGRP release was lost whereas the PGE2 release was unchanged. Thus, neither nerve endings nor neuropeptides contribute essentially to BK-induced PGE2 release in the skin. However, excessive neuropeptide levels, as under NKI blockade facilitate PGE2 formation, which may play a role in sustained inflammation. PMID- 10674472 TI - Amyloid Abeta40 CSF concentrations correlate to frontal lobe atrophy in frontotemporal dementia. AB - We wanted to further study amyloid Abeta protein alterations in non-AD neurodegenerative diseases. Cerebrospinal fluid concentrations of the amyloid Abeta protein with 40 (Abeta40) and 42 (Abeta42) amino acid residues were measured in eleven patients with frontotemporal dementia (FTD). Abeta40 and Abeta42 concentrations were related to the degree of frontal lobe atrophy as assessed with MRI volumetry. Abeta40 concentrations showed a statistically significant linear correlation with degree of frontal lobe atrophy (r = -0.77, p<0.02). Similar results have not been found in previous studies of CSF Abeta40 concentrations and atrophy in patients with AD which suggest that the role of Abeta40 differs between the pathological processes of FTD and AD. PMID- 10674473 TI - Contralateral treatment with xylocaine reduces nociceptive behaviour in mononeuropathic rats. AB - The aim of the present study was to establish whether contralateral treatment with local anaesthetics reduces nociceptive behaviour in mononeuropathic rats. Contralateral treatment with xylocaine on day 6 and 11 following sciatic nerve ligation increased withdrawal latencies to thermal but not to mechanical stimulation for 3-4 days. Rats who received contralateral treatment with xylocaine on day 6 and 11 showed a reduced autotomy score during the following 6 weeks. To compare ipsilateral and contralateral effects another set of experiments was performed. Rats treated contralaterally on day 11 had a significantly lower autotomy score at week 4 and 6. Our results demonstrate that contralateral treatment with xylocaine reduces nociceptive behaviour in mononeuropathic rats for days or weeks. PMID- 10674474 TI - The CCTTT polymorphism in the NOS2A gene is associated with dementia with Lewy bodies. AB - We report the analysis of the allele distribution of a (CCTTT)n pentanucleotide repeat within the promoter region of the NOS2A gene in DNA samples from patients with autopsy confirmed Alzheimer's disease (AD) and dementia with Lewy bodies (DLB) type. A significant difference was observed in the allelic distribution between the control group and the DLB group (chi2 = 15.175, df = 5; p<0.01), with an increased occurrence of the eight and nine repeat alleles, and a marked under representation of the 11 repeat allele. Genotype frequencies in the DLB group also differed significantly from controls (p<0.012). These results suggest that variations in the NOS2A gene may predispose to the development of DLB. PMID- 10674475 TI - c-fos expression and redox state of cytochrome oxidase of rat brain in hypoxia. AB - Hypoxic induction of c-fos was studied in rat brains as a function of the cerebral oxygenation state using near-infrared spectroscopy by which the hemoglobin oxygenation state and redox state of mitochondrial cytochrome oxidase could be monitored noninvasively. Following reoxygenation after hypoxia, the expression of c-fos and MAP2 mRNAs was followed by reverse transcription-coupled PCR. The expression of MAP2 remained unchanged throughout all the conditions from 21 to 8% FiO2. Under mildly hypoxia conditions, c-fos mRNA was not induced. Hemoglobin was partially deoxygenated but cytochrome oxidase remained fully oxidized. Severe hypoxia, where cytochrome oxidase was reduced, caused a significant induction of c-fos mRNA At this stage, the oxygen concentration in cerebral tissue fell to < 10(-7) M. These data suggest that the decline in oxidative phosphorylation might be a trigger for the induction of c-fos mRNA. PMID- 10674476 TI - Analysis of splicing of four mouse JNK/SAPKalpha variants. AB - The JNK/SAPK (c-Jun NH2-terminal kinase/stress-activated protein kinase) cascade is activated by a variety of stress stimuli and by the inflammatory cytokines interleukin-I (IL-I) and tumor necrosis factor alpha (TNFalpha). Four splice variants of the mouse JNK/SAPKalpha isoform, which differ in a region located in subdomains IX-X of the protein, were previously identified. Analysis of the sequence of the central region of the mouse JNK/SAPKalpha gene indicates that splice variants I and II are generated by a typical alternative splicing mechanism, while splice variants III and IV are generated by a less common mechanism, where alternative 3' splice sites located inside an exon (cryptic sites) are selected. The major splice variants alphaI and all have a wide and similar distribution in hippocampus, cerebral cortex, caudate-putamen, amygdala and the granule cell layer of cerebellum, although their expression is specifically regulated in certain cell types. PMID- 10674477 TI - Quantitative PCR analysis of AMPA receptor composition in two paradigms of global ischemia. AB - Quantitative PCR was used to analyse the expression of GluR1, GluR2, GluR2 flip, GluR2 flop and GluR3 mRNA in animals after ischemia and tolerance-inducing ischemia. The ischemic animals showed a decrease in the GluRs to approximately 30%, except for GluR2-flip, which decreased to 75%. The tolerance animals displayed regulation of GluR1 to 75%, GluR2 and GluR2-flop to 283% and 265% respectively. We did not find a correlation between GluR2 regulation and cell loss in the ischemic group. The selective upregulation of GluR2/GluR2 flop in tolerant animals indicates a possible mechanism for enhanced AMPA receptor desensitisation leading to tolerance to ischemia. PMID- 10674478 TI - Ultradian rhythmic neuronal oscillation in the intergeniculate leaflet. AB - Our paper is the first to describe ultradian rhythmic neuronal oscillation in the intergeniculate leaflet (IGL) of the rat. We recorded a multiple-unit neuronal activity (MUA) from dorsal to ventral parts of the lateral geniculate nucleus (LGN) in anaesthetized rats. In all the subdivisions of the lateral geniculate complex we observed spontaneous irregular firing rates of cells. However only at the anatomical localisation of the IGL, after the light was on, those responses exhibited burst firing with a constant interburst interval, which lasted several hours until the light was off. The duration of that rhythmic oscillation obtained by means of Fourier's analysis was approximately 124 s. To date we have not had sufficient data to discuss possible mechanisms of this neuronal rhythmicity. We can only conclude that light is the most important stimulus not only for suprachiasmatic nuclei (SCN), but also for the IGL. On the other hand, we can neither exclude nor confirm that in order to evoke ultradian rhythmical oscillation in the IGL, in addition to light also non-photic information is necessary. PMID- 10674479 TI - Retention of words in long-term memory: a functional neuroanatomical study with PET. AB - We used PET to identify brain regions associated with retention of verbal materials in long-term memory. During a PET scan, subjects repeated many sets of words one after another. In a retention condition, they were simultaneously required to retain 10 key words that were irrelevant to the repetition task. Significant increases in regional cerebral blood flow during the retention condition were found in bilateral parahippocampal regions, the left prefrontal and parietal association cortices, the supplementary motor area, the neostriatum and the cerebellum. We clearly demonstrated that retention of verbal materials was accompanied by neural activities in the medial temporal lobes. We also showed that, in the early phase, retention of words in long-term memory recruited left cortical areas surrounding those relevant to verbal short-term memory. PMID- 10674480 TI - Lateralization of movement-related potentials and the size of corpus callosum. AB - Using structural MRI and whole-head EEG recordings, we analyzed the correlations between the anatomical parameters of the corpus callosum and the hemispheric distribution of the cortical movement-related potentials during right finger and shoulder movements in nine right-handed men. Statistically significant correlation was found only in finger movements. A relatively large genu and the anterior part of the truncus of the corpus callosum correlated with enhanced pre movement EEG potential over the ipsilateral M1/S1 area. The lateralization of the movement-related potentials correlates with the size of those callosal regions which connect the homologous areas of the primary sensorimotor and frontal cortices. PMID- 10674481 TI - ERP development in the rat in the course of learning two-tone discrimination task. AB - To clarify some neurophysiological aspects of learning, we investigated the relationship between the course of learning and development of ERP and investigated developmental processes of ERPs. Nine male Sprague-Dawley rats were trained for a two-tone discrimination task and rat P3 and N1 component were longitudinally recorded. Both rat P3 and N1 gradually increased with learning only for target tones. An improvement in the proportion of correct responses preceded the increase in ERPs, and the increase in P3 and N1 proceeded almost simultaneously. These findings suggest that multiple kinds of information processing were acquired with learning the two-tone discrimination task. ERP development could be utilized as an index of establishment of learning. PMID- 10674482 TI - The selectivity of the occipitotemporal M170 for faces. AB - Evidence from fMRI, ERPs and intracranial recordings suggests the existence of face-specific mechanisms in the primate occipitotemporal cortex. The present study used a 64-channel MEG system to monitor neural activity while normal subjects viewed a sequence of grayscale photographs of a variety of unfamiliar faces and non-face stimuli. In 14 of 15 subjects, face stimuli evoked a larger response than non-face stimuli at a latency of 160 ms after stimulus onset at bilateral occipitotemporal sensors. Inverted face stimuli elicited responses that were no different in amplitude but 13 ms later in latency than upright faces. The profile of this M170 response across stimulus conditions is largely consistent with prior results using scalp and subdural ERPs. PMID- 10674483 TI - Ovarian steroids and raloxifene prevent MPTP-induced dopamine depletion in mice. AB - The activity of steroids was studied in 1-methyl-phenyl-1,2,3,6 tetrahydropyridine (MPTP) lesioned retired breeder C57BL/6 male mice as a model of Parkinson's disease. Steroids were injected daily for 5 days before MPTP (4 injections, 15 mg/kg i.p., at 2 h intervals) and hormonal treatment continued for 5 more days. Mice that received 17beta-estradiol or progesterone or raloxifene (a selective estrogen receptor modulator) and MPTP had striatal concentrations of dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) similar to those in control animals, whereas mice that received MPTP alone or with 17alpha-estradiol (the isomer with weak estrogenic activity) had an extensive decrease of DA and its metabolites. These results suggest stereospecific prevention of MPTP-induced dopamine loss by 17beta estradiol, which is also observed with progesterone and raloxifene. PMID- 10674484 TI - Preserved olfactory short-term memory after combined cholinergic and serotonergic lesions using 192 IgG-saporin and 5,7-dihydroxytryptamine in rats. AB - Young adult Long-Evans female rats were subjected to intracerebroventricular injections of 150 microg 5,7-dihydroxytryptamine (5,7-DHT), 2 microg 192 IgG saporin, or a combination of both neurotoxins. All rats were tested for olfactory recognition (short-term memory) using a task based on spontaneous exploration of odor sources. Compared with animals undergoing sham operations, 5,7-DHT reduced the concentration of serotonin by 60-80% in the frontoparietal cortex, hippocampus, striatum and the olfactory bulbs. After 192 IgG-saporin treatment, acetylcholine concentrations were reduced by approximately 40% in all these structures, except the striatum. Neither lesion induced a significant deficit in olfactory recognition. These data suggest that combined lesions of cholinergic and serotonergic neurons in the rat brain do not alter olfactory perception or olfactory short-term memory. PMID- 10674485 TI - Reversible lesions of the rhinal cortex produce delayed non-matching-to-sample deficits in rats. AB - Rats with cannulae guides implanted in the rhinal cortex were tested on a delayed non-matching-to-sample task, following either lidocaine or sham microinfusions. Bilateral lidocaine microinfusions to the rhinal cortex produced significant delayed non-matching-to-sample deficits. These results are consistent with the putative role of the rhinal cortex in object recognition but because the deficits were not shown to be time dependent, non-mnemonic interpretations cannot be ruled out. These results also illustrate the utility of reversible lidocaine lesions in the study of the neuroanatomical basis of delayed non-matching-to-sample. PMID- 10674486 TI - Acute treatment of hypertension increases infarct sizes in spontaneously hypertensive rats. AB - We studied the effect of dihydralazine treatment of hypertension in spontaneously hypertensive stroke-prone rats in a model of permanent focal cerebral ischemia (stroke). After occlusion of the middle cerebral artery systemic arterial pressure (SAP) was lowered with a computer controlled infusion device from 163 to 135 or 117 mm Hg for 24h. In the control group SAP was not manipulated. Reduction of SAP to normotension (117 mm Hg) significantly worsened outcome and increased infarct volume measured 7 days after induction of ischemia, whereas a mild reduction of SAP (to 137 mm Hg) had no statistically significant effect on outcome or infarct volume. We conclude that pharmacological treatment of hypertension may negatively affect neurological outcome and infarct volume in a rat stroke model. PMID- 10674487 TI - AlphaB-crystallin regulates intermediate filament organization in situ. AB - AlphaB-crystallin is a small heat shock protein (hsp) and molecular chaperone that can interact with a wide spectrum of cellular components including intermediate filaments (IF). The significance of these interactions is not currently known. We have tested whether increased alphaB-crystallin expression effects changes in the IF systems in situ. Adenoviral-mediated gene transfer was used to overexpress alphaB-crystallin in primary astrocytes. A positive correlation was observed between overexpression of alphaB-crystallin and diffuse, filigree IF. AlphaB-crystallin did not appear to alter the polymerization state of IF proteins. These data show that an increase in alphaB-crystallin expression in the absence of stress can modify the organizational state of IF and that alphaB-crystallin can function as an IF debundling protein. PMID- 10674488 TI - Auditory responses from the frontal cortex in the mustached bat, Pteronotus parnellii. AB - Response properties of neurons in an auditory field in the frontal cortex of the mustached bat, Pteronotus parnellii, have not been studied before. We recorded neural responses to constant frequency (CF) stimuli from the frontal auditory field in awake animals. The majority (75%) of neurons in this area responded well and often exhibited low thresholds to CF stimuli. Most CF-responsive neurons exhibited sharp tuning with values of > 180 for Q10db, a quality factor expressing the sharpness of tuning at 10dB above threshold. Neurons at 13 recording sites exhibited combination sensitivity in that their responses were facilitated by presenting combinations of either CF1/CF2 and/or CF1/CF3 components of the mustached bat's echolocation signal. Unlike the typical on responses to a 30 ms tone, observed in the mustached bat's auditory cortex and at subcortical levels, many frontal auditory neurons exhibited loosely time locked firing patterns that lasted for > 100 ms. PMID- 10674489 TI - The 68K protease has beta-secretase-like activity for lymphocyte precursor protein but not for brain substrate. AB - Processing and metabolism of beta-amyloid precursor protein (APP) and generation of a variety of beta-amyloid (Abeta) peptides in the human brain is essentially associated with pathophysiology of Alzheimer's disease (AD). APP degradation activity of the 68 kDa serine protease, which was originally prepared from familial AD lymphoblastoid cells and harbors beta-secretase-like activity, was analyzed by Western blot using anti Abeta 1/40 antibody and anti APP cytoplasmic domain (CT) antibody. Native lymphocyte APP (LAPP) prepared from normal or AD derived lymphoblastoid cells was degraded by the protease, generating a 16 kDa Abeta-bearing C-terminal fragment of APP. N-terminal amino acid sequencing of the fragment indicated that the protease cleaves LAPP at the Abeta-N-terminus. When the LAPP was treated with chondroitinase ABC prior to proteolysis, the activity to generate the fragment was inhibited, but pretreatment with heparitinase resulted in no effect. Native hippocampal APP prepared from normal brain, however, did not generate the 16 kDa peptide by the protease treatment. These results suggest that the process of APP degradation and Abeta-peptides generation, including beta-secretase activity, is associated with tissue specificity of both APP substrate and proteases. They also indicate that sulfated glycoconjugates attached to a portion of APP isoforms may play a role as a molecular determinant in the proteolysis. PMID- 10674490 TI - Brainstem microstimulation activates sympathetic fibers in pudendal nerve motor branch. AB - A bilateral spino-bulbo-spinal circuit conveys information from/to the male urogenital tract and perineal muscles. This is the first electrophysiological report of another descending pathway, one which conveys output from the medullary reticular formation (MRF) to activate postganglionic sympathetic fibers contained within the motor branch of the pudendal nerve (PudM). In anesthetized rats, long latency (> 150 ms) discharges were elicited in the PudM following ipsilateral or contralateral microstimulation of the MRF. These firing bursts were not observed in rats after sectioning the lower lumbar sympathetic trunk. The most robust activation was observed when neurons in or near the lateral paragigantocellular reticular nucleus were microstimulated bilaterally. Urogenital dysfunction that occurs following severe spinal cord injury probably results from disrupting these and other supraspinal circuits. PMID- 10674491 TI - Hyperinsulinemia increases norepinephrine metabolism in the ventromedial hypothalamus of rats. AB - Numerous studies have implicated increased ventromedial hypothalamic (VMH) norepinephrine (NE) activity as a contributing factor to the obese, hyperinsulinemic, glucose intolerant condition. However, factors contributing to the increased VMH NE activity remain unknown. This study therefore investigated in normal rats the effect of a hyperinsulinemic-euglycemic clamp on VMH monoamine turnover and utilization via simultaneous VMH microdialysis to establish a role for hyperinsulinemia in the stimulation of VMH NE activity. Within 20 min of initiation of the hyperinsulinemic-euglycemic clamp, VMH extracellular methoxyhydroxy phenylglycol (metabolite of NE) level increased by 54% and remained approximately at this level for the 100 min duration of the clamp relative to control values (p<0.05). Hyperinsulinemia did not affect VMH dopamine or serotonin metabolism. Subsequent establishment of a hyperinsulinemic hypoglycemic camp did not alter the VMH monoamine metabolism profile relative to the hyperinsulinemic-euglycemic clamp. Infusion of saline (as control) in a separate group of rats over the entire clamp period induced no changes in any monoamine metabolic profile relative to baseline. Hyperinsulinemia can feedback to stimulate VMH NE activity and, as a result, may contribute to the initiation and/or perpetuation of the obese, hyperinsulinemic, glucose-intolerant state. PMID- 10674492 TI - Effects of retinoic acid and tumor necrosis factor alpha on GL-15 glioblastoma cells. AB - Glioblastomas are particularly resistant to classical antitumor treatments. Retinoids, which proved effective in the treatment of promyelocytic leukemia, have been used for clinical assays on glioma tumors with only moderate effects; however in some cases they were active in combination with another therapy. These observations prompted us to analyse the efficacy of combining retinoic acid (RA) with a cytokine on a clonal human glioma cell line. On GL-15 cells, RA and tumor necrosis factor alpha (TNFalpha) both reduced the glial fibrillary acidic protein level and DNA synthesis and induced apoptotic pathways, but they were significantly more effective when used together. The up-regulation of the p55 TNF receptors observed during RA exposure might explain this cooperative effect. PMID- 10674493 TI - Kainate receptors activate NF-kappaB via MAP kinase in striatal neurones. AB - The transcription factor NF-kappaB has been implicated in the synaptic plasticity and neurotoxicity mediated by ionotropic glutamate receptors in the striatum. However, the class of glutamate receptor and the intracellular pathways involved have not been determined. Kainate, but not AMPA or NMDA, was found to activate NF kappaB in superfused slices of rat striatum. A similar activation was produced by the calcium ionophore A23187. The NF-kappaB activation by kainate was not observed in the absence of extracellular calcium, and was blocked by the p44/p42 MAP kinase inhibitor PD98059, but not by the p38 MAP kinase inhibitor SB203580. This demonstrates that striatal kainate receptors are coupled to NF-kappaB activation via calcium influx and p44/p42 MAP kinase activation. PMID- 10674494 TI - Sensory-specific satiety-related olfactory activation of the human orbitofrontal cortex. AB - When a food is eaten to satiety, its reward value decreases. This decrease is usually greater for the food eaten to satiety than for other foods, an effect termed sensory-specific satiety. In an fMRI investigation it was shown that for a region of the orbitofrontal cortex the activation produced by the odour of the food eaten to satiety decreased, whereas there was no similar decrease for the odour of a food not eaten in the meal. This effect was shown both by a voxel-wise SPM contrast (p<0.05 corrected) and an ANOVA performed on the mean percentage change in BOLD signal in the identified clusters of voxels (p<0.006). These results show that activation of a region of the human orbitofrontal cortex is related to olfactory sensory-specific satiety. PMID- 10674495 TI - Presenilin I expression in yeast lowers secretion of the amyloid precursor protein. AB - Presenilin (PS) mutations are associated with early-onset Alzheimer's disease and PS proteins are involved with gamma-secretase cleavage of the amyloid precursor protein, APP. We have shown previously that alpha-, beta- and gamma-secretase cleavages of APP are conserved in Pichia pastoris. Here, we report co-expression of APP and PSI in P. pastoris and show by immunoelectron microscopy colocalization of these two proteins in expanded endoplasmic reticulum. Western blot analysis indicates a drastic reduction of both alpha- and beta-secretase products. A relative increase in beta-secretase product derived from immature APP is also observed, pointing to a beta-secretase activity of P. pastoris associated with the early secretory pathway. PMID- 10674496 TI - TNF-alpha induced over-expression of GFAP is associated with MAPKs. AB - Increased levels of tumor necrosis factor-alpha (TNF-alpha), a pluripotent cytokine that is reportedly mitogenic to astrocytes, are associated with the expression of glial fibrillary acidic protein (GFAP), the most specific marker for astrocytes, in many neuropathological conditions, including brain injury, CNS infection, Creutzfeldt-Jakob disease and Alzheimer's disease. Here, we show that treatment of cultured astrocytes with TNF-alpha resulted in dramatic over expression of GFAP, associated with a substantial activation of the mitogen activated protein kinase (MAPK) Erk2 (extracellular signal-regulated protein kinase). We also demonstrate that TNF-alpha-induced over-expression of GFAP was significantly attenuated by the MAPK inhibitor PD98059. We conclude that TNF alpha may upregulate GFAP through the MAPK signaling pathway. Because increased GFAP is a hallmark of reactive gliosis, understanding the mechanisms that regulate GFAP expression may facilitate development of strategies to minimize the gliosis associated with many brain diseases. PMID- 10674497 TI - Effects of 902 MHz electromagnetic field emitted by cellular telephones on response times in humans. AB - The present study examined possible influences of a 902 MHz electromagnetic field emitted by cellular telephones on cognitive functioning in 48 healthy humans. A battery of 12 reaction time tasks was performed twice by each participant in a counterbalanced order: once with and once without the exposure to the field. The results showed that the exposure to the electromagnetic field speeded up response times in simple reaction time and vigilance tasks and that the cognitive time needed in a mental arithmetics task was decreased. The results suggest that exposure to the electromagnetic field emitted by cellular telephones may have a facilitatory effect on brain functioning, especially in tasks requiring attention and manipulation of information in working memory. PMID- 10674498 TI - Calcium channels functional roles in the frog semicircular canal. AB - Different types of voltage-operated calcium channels have been described in hair cells; however, no clear functional role has been assigned to them. As a first functional characterization of vestibular calcium channels, we studied the effect of several calcium channel agonists and antagonists on whole nerve firing rate in an isolated frog semicircular canal preparation. Resting activity was affected by all dihydropyridines tested and by omegaconotoxin GVIA, whereas only nimodipine was able to reduce the mechanically evoked activity. These results indicate that nimodipine-sensitive channels play a major role in afferent transmitter release, and omega-conotoxin GVIA sensitive channels regulate the afferent firing (possibly on the postsynaptic side) but with a less important role. PMID- 10674499 TI - Auditory responses from the frontal cortex in the short-tailed fruit bat Carollia perspicillata. AB - Based on neuroanatomical findings it was hypothesized that an area in the bat frontal cortex is part of a sensorimotor feedback loop and probably important to goal-directed behaviors guided by auditory information. The present report describes the basic stimulus preferences and response properties of neurons from this area in the short-tailed fruit bat Carollia perspicillata. Responses to acoustic stimuli mimicking biosonar pulse-echo (i.e. FM-FM) combinations were found to be facilitated throughout but only rarely exhibited tuning to pulse-echo delay. As opposed to the often sharply delay-tuned FM-FM neurons in the species' auditory cortex, frontal cortical FM-FM neurons seem to be suited for indicating the presence of an insonified object irrespective of its distance and hence are likely to function as novelty detectors and to trigger changes in the bats' orientation behavior. PMID- 10674500 TI - Neuroprotection achieved with a novel proteasome inhibitor which blocks NF-kappaB activation. AB - Activated NF-kappaB contributes to cerebral infarction by triggering a neuro inflammatory response. Rats subjected to 90min middle cerebral artery occlusion developed a cortical infarct of 20+/-4% of hemispheric volume (n = 8). Treatment with the proteasome inhibitor CVT-634 resulted in a significantly smaller infarct of 13+/-2% (n = 7, p<0.01) and 12+/-2% (n = 8, p<0.001) of hemispheric volume at 1 day and 7 days, respectively. Since regional cerebral blood flows for the core and penumbral regions were not affected, we concluded that all animals received the same ischemic insult The reduction in infarction persisted for 7 days. This is the first indication that a proteasome inhibitor can reduce infarct volume in a focal model of cerebral ischemia. PMID- 10674501 TI - Anesthesia human resources in Canada. PMID- 10674502 TI - Litigation in Canada against anesthesiologists practicing regional anesthesia. A review of closed claims. AB - PURPOSE: To review the pattern of malpractice litigation related to regional anesthesia in Canada. SOURCE: The Canadian Medical Protective Association (CMPA) provided with information about all anesthesia claims that closed in the years 1990-1997. PRINCIPAL FINDINGS: In the period 1990-97 there were 7,909 closed legal actions involving all CMPA members (56,000). Of these, there were 310 cases involving anesthesiologists, of which 61 cases (approximately 20%) were related to regional anesthesia. Forty-two involved neuraxial blocks, and the legal outcome was favourable (dismissed or judgement in favour of the defendant doctor) in 37 claims. Nineteen claims involved peripheral nerve blocks. All these had favourable legal outcomes. Overall, 10% of regional anesthesia claims have unfavourable outcomes, compared with 28% of all anesthesia related claims and 30% of all CMPA members' claims. The degree of disability in the regional anesthesia claims were: none 10%; minor 49%; major 36%; catastrophic 5%. There were no deaths in the malpractice claims involving regional anesthesia, compared with 17% in the all anesthesia group and 11% in all members' claims. CONCLUSION: Twenty percent of all anesthesia claims in Canada are related to regional anesthesia. The legal outcome of these claims is favourable in 90%. Unfavourable clinical outcome is associated with catastrophic or major injury. There were no deaths in the regional anesthesia claims. PMID- 10674503 TI - A comparison of patient-controlled analgesia fentanyl and alfentanil for labour analgesia. AB - PURPOSE: To determine the analgesic efficacy of equipotent doses of PCA (patient controlled analgesia) fentanyl and PCA alfentanil for labour pain. METHODS: Twenty three, ASA I - II parturients between 32-42 wk gestational age in whom epidural analgesia was contraindicated were randomized to receive PCA fentanyl (Group F)or alfentanil (Group A). Plain numbered vials contained 21 ml fentanyl 50 microg x ml(-1) or alfentanil 500 microg x ml(-1). A one millilitre loading dose was administered. The PCA solution was prepared by diluting 10 ml study drug with 40 ml saline and the PCA pump was programmed to deliver a dose of 2 ml, delay of five minutes and a basal rate of 2 ml x hr(-1). Maternal measurements obtained were hourly drug dose, total dose, Visual Analog Pain Score (VAPS) q 30 min, sedation score q 1 hr and side effects. Neonates were assessed by 1,5, and 10-min Apgar scores, umbilical venous and arterial blood gases and neurobehavioural scores at four and 24 hr. RESULTS: Mean VAPS from 7 - 10 cm cervical dilatation were higher in Group A than in Group F (85.7+/-13.9 vs. 64.6+/-12.1; P<0.01) There were no inter-group differences in VAPS from 1-3 cm, or from 4-6 cm dilatation, in maternal sedation scores or side effects, or in neonatal outcomes. CONCLUSION: In the doses prescribed in this study, PCA fentanyl was found to provide more effective analgesia in late first stage labour than PCA alfentanil. PMID- 10674504 TI - Plasma concentration of flumazenil following intranasal administration in children. AB - PURPOSE: A pharmacokinetic study in children to determine plasma flumazenil concentrations after the intranasal administration of 40 microg x kg(-1). METHODS: Following institutional approval and informed written consent, 11 ASA physical status I-II patients, aged two to six years, undergoing general anesthesia for dental surgery were recruited. After induction, 40 microg x kg(-1) flumazenil Anexate, Roche, 0.1 mg x mL(-1) (0.4 mL x kg(-1))) were administered via a syringe as drops, prior to nasal intubation. Venous plasma samples were drawn prior to administration of flumazenil (t = 0), and then at 2, 4, 6, 8, 10, 15, 20, 30, 40, 60, and 120 min thereafter. The plasma samples were immediately processed by the on-site laboratory and then stored at -70 degrees C, before batch analysis via high performance liquid chromatography assay. Pharmacokinetic data calculations were performed using WinNonLin software (Scientific Consulting Inc.). RESULTS: Eleven patients were studied, but data for one patient were discarded due to insufficient sampling. The median age was 4.3 yr (range 3 to 6), with a median weight of 18.9 kg (range 14.9 to 22.2). There were seven boys and three girls. Mean Cmax was 67.8 ng x mL(-1) (SD 41.9), with Tmax at two minutes. The calculated half-life was 122 min (SD 99). CONCLUSION: The mean plasma concentrations of flumazenil attained were similar to those reported after intravenous administration, and may be sufficient to antagonize the side-effects of benzodiazepines. This route of administration may be useful when the intravenous route is not readily available. PMID- 10674505 TI - Rate of change of cerebral blood flow velocity with hyperventilation during anesthesia in humans. AB - PURPOSE: Although it has been suggested that the rate at which the cerebral circulation responds to changes in PaCO2 is different with differing anesthetics, there have been no attempts to measure this. Transcranial Doppler allows the continuous measurement of cerebral blood flow velocity (CBFV) and any changes over time. Our aim was to compare the rate of change of CBFV when end-tidal CO2 (P(ET)CO2) was rapidly altered during halothane or isoflurane anesthesia. METHODS: Twenty-eight unpremedicated healthy patients were randomly assigned to receive air/O2 and either 1-1.5 MAC halothane or isoflurane as the primary anesthetic. After 15 min of steady state, P(ET)CO2 was rapidly reduced from 45 mm Hg to 30 mm Hg. CBFV and P(ET)CO2 were recorded every 30 sec for the next 10 min. RESULTS: The rate of change of normalized CBFV (delta CBFV vs. delta time) was more rapid in the isoflurane group (P <0.0001) especially in the initial few minutes. In all patients anesthetized with isoflurane, and in all but two patients anesthetized with halothane, the reduction in P(ET)CO2 produced a corresponding decrease in CBFV However, there were no differences in the magnitude of cerebrovascular CO2 reactivity (delta CBFV vs. delta P(ET)CO2) between the two groups. CONCLUSIONS: The rate of change of CBFV was faster in the isoflurane than in the halothane group especially in the initial few minutes. Indeed, for two patients in the halothane group Vmca did not change despite a change in P(ET)CO2. This may be of clinical importance when cerebrovascular tone needs to be changed rapidly. PMID- 10674506 TI - Cerebral oxygenation is better during mild hypothermic than normothermic cardiopulmonary bypass. AB - PURPOSE: Normothermic cardiopulmonary bypass (CPB) has been recently used in cardiac surgery. However, there is a controversy whether there is a difference in incidence of neurological disorder after coronary artery bypass graft (CABG) surgery between normothermic CPB and mild hypothermic CPB. In this study, we assessed the effects of normothermia and mild hypothermia (32 degrees C) during CPB on jugular oxygen saturation (SjvO2). METHODS: Twenty patients scheduled for elective CABG surgery were divided into two groups. Group 1 (n = 10) underwent normothermic (>35 degrees C) CPB, and Group 2 (n = 10) underwent mild hypothermic (32 degrees C) CPB. Alpha-stat blood gas regulation was applied. After inducing anesthesia, a 4.0 French fibre optic oximetry oxygen saturation catheter was inserted into the right jugular bulb to monitor SjvO2 continuously throughout anesthesia and surgery. RESULTS: The SjvO2 in the normothermic group was decreased at 20 (41.5+/-2.4%) and 40 min (43.8+/-2.8%) after the onset of CPB compared with control (53.9+/-5.4%, P<0.05). However, there was no change in SjvO2 in the mild hypothermic group during the study. No changes in jugular venous-arterial differences of lactate or creatine phosphokinase isoenzyme BB were observed in two groups during the study. CONCLUSIONS: Cerebral oxygenation, as assessed by SjvO2 was increased during mild hypothermic CPB than during normothermic CPB. PMID- 10674507 TI - Comparison of differential blockade during spinal anesthesia using isobaric vs. hyperbaric lidocaine 2%. AB - PURPOSE: To compare the extent of the sensory, motor and sympathetic block produced by a single dose of 60 mg lidocaine at the same concentration (2%) and volume but at different baricity injected intraspinally. METHOD: In a randomised double blind study, 40 ASA I-II patients were scheduled for elective surgery (orthopedic, urologic, peripheral vascular and lower digestive procedure). They were divided in two groups. Twenty patients received 60 mg lidocaine 2% in a hyperbaric solution and 20 received 60 mg lidocaine 2% in a isobaric solution. The levels of sensory (pinprick, ice) motor (Bromage scale) and sympathetic blockade (galvanometry, cutaneous blood flow, temperature) were measured at 0, 5, 10, 15, 20 and 30 min. RESULTS: There were no differences between the groups with regard to maximal height of sympathetic block, sensory level to pinprick: T5 +/ 2.4 for isobaric group, T6 +/-3.6 for hyperbaric group or to cold: T3 +/-2.3 for isobaric group, T4 +/-2.7 for hyperbaric group. Hyperbaric lidocaine 2% produced a more pronounced sensory (pinprick, ice) and motor block on the dependant than on the non-dependant side. CONCLUSION: The baricity of 60 mg lidocaine injected intraspinally in the lateral decubitus position did not influence the cephalad spread of sensory or sympathethic blockade. In the hyperbaric group, the dependent side showed a more pronounced sensory (pinprick, ice), and motor block. PMID- 10674508 TI - Neuromuscular effects of rapacuronium in pediatric patients during nitrous oxide halothane anesthesia: comparison with mivacurium. AB - PURPOSE: To describe neuromuscular effects of rapacuronium in pediatric patients during N2O-halothane anesthesia and compare them with mivacurium in children. METHODS: 103 pediatric patients, seven days -12 yr, received rapacuronium or mivacurium during N2O-halothane anesthesia. Onset and recovery of block were measured using EMG (Datex). Block was compared between groups based on drug treatment and age. Children < two years received 1 or 2 mg x kg(-1) rapacuronium: 2-12 yr received either 2 mg x kg(-1) or 3 mg x kg(-1) rapacuronium, or 0.2 mg x kg(-1) mivacurium. RESULTS: There were no differences in onset (1.7+/-1.8 min) or maximum block (T1 2.4+/-8%) among neonates, infants, and toddlers after either dose of rapacuronium. There was no difference between 1 and 2 mg x kg(-1) of rapacuronium block at 60 sec. Train-of-four ratio (T4/T1) >0.7 occurred later after 2 mg x kg(-1) than 1 mg x kg(-1) in these patients (P<0.05). There was no difference in T25 among neonates, infants and toddlers for 1 mg x kg(-1) or 2 mg x kg(-1) doses. Rapacuronium, 3 mg x kg(-1), produced maximum block 1.5 min earlier than did mivacurium, 0.2 mg x kg(-1) (P<0.001). There was no difference in block at 60 sec, maximum block or time to maximum block between 2 and 3 mg x kg(-1) rapacuronium for children > two years of age. Maximum block occurred 1.0+/ 0.5 min after 2 or 3 mg x kg(-1) when T1 was 0.2+/-1.1% of baseline. T25 and T4/T1 >0.7 occurred 10 to 11 min later after this dose of rapacuronium than after mivacurium. CONCLUSION: Rapacuronium produces block earlier than mivacurium. Recovery from rapacuronium block is dose related and slower than that following mivacurium during halothane anesthesia. PMID- 10674509 TI - Accuracy of carboxyhemoglobin dilution method for the measurement of circulating blood volume. AB - PURPOSE: The management of circulating blood volume (BVc) is crucial in intensive care unit (ICU) patients. The purpose of this study was to verify the accuracy and precision of the carbon monoxide-labeled hemoglobin (CO-Hb) dilution method (CO method) by comparing it with the 51Cr-labeled erythrocyte dilution method (51Cr method) for the measurement of BVc. METHODS: A prospective study was performed in 18 patients who underwent coronary artery bypass grafting (CABG) under mild hypothermic cardiopulmonary bypass (CPB). The BVc was measured by both the CO method and the 51Cr method at 24 hr after ICU admission in order to verify the accuracy and precision of the CO method. Paired data were assessed in absolute terms, and percentage errors were calculated by the degree of agreement. RESULTS: Small mean differences and standard deviations between the CO method and the 51Cr method (-70.2 +/-184.8 mL) and small percentage errors (-0.49+/-1.29%) indicated the accuracy and precision of the CO method, and a close correlation was observed (r = 0.97). CONCLUSION: The CO method can measure BVc with a similar degree of accuracy as the 51Cr method. It is simple, repeatable and safe without the risk of exposure to radioactivity in the ICU. PMID- 10674510 TI - Epidural meperidine does not cause hemodynamic changes in the term parturient. AB - PURPOSE: Meperidine has local anesthetic properties and, therefore, when given epidurally it has the potential to cause hemodynamic changes. Our objective was to study the hemodynamic effects of an analgesic dose of epidural meperidine (50 mg) in 34 ASA 1-2 term parturients scheduled for elective Cesarean section under epidural anesthesia. METHODS: A lumbar epidural catheter was inserted and patients lay in the supine left wedge position. Intravenous fluid preload was withheld, and hemodynamic measurements comprising of mean arterial pressure, cardiac output and heart rate were made using automatic oscillotonometry (Dinamap 1486SX) and transthoracic electrical bioimpedance (Bomed NCCOM3). Following baseline measurements, the hemodynamic effects of sequential epidural injection of first, 10 ml saline, and 20 min thereafter, 50 mg meperidine diluted to 10 ml with saline, were recorded. Sensory blockade was assessed following each injection using loss of temperature discrimination to ice. Paired Student t tests were used to compare changes in hemodynamic variables. RESULTS: Epidural meperidine produced a small increase from the saline values in the mean (SD) cardiac output of 5.81 +/-1.44 to 6.04+/-1.54 L x min(-1) (P<0.05), and mean arterial pressure of 77.1+/-8.8 to 79.3+/-9.9 mm Hg (P<0.05). Sensory changes, the upper level of which ranged from L1 to T1, were detected in 94% of patients given epidural meperidine. Epidural saline injection had no such hemodynamic effects, but produced a detectable sensory level in two patients. CONCLUSION: Epidural meperidine, 50 mg, caused minimal hemodynamic changes in term parturients. PMID- 10674511 TI - Familial hypokalemic periodic paralysis and Wolff-Parkinson-White syndrome in pregnancy. AB - PURPOSE: To describe the anesthetic and obstetrical management of a pregnant patient with co-existing Familial Hypokalemic Periodic Paralysis (FHPP) and Wolff Parkinson-White syndrome (WPW). CLINICAL FEATURES: A 29 yr-old primigravida with FHPP and WPW presented to the antenatal clinic at 18 wk gestation, for consideration of her anesthetic and obstetrical management during labour and delivery. A plan was constructed to avoid the known precipitating factors of FHPP including carbohydrate loading, cold, mental stress and exercise, which could lead to acute attacks of weakness. She presented for induction of labour at 41 wk and three days. An epidural catheter was sited early in labour. The second stage was limited to less than one hour. She had a rotational forceps delivery for which the epidural was extended to provide anesthesia. A healthy male baby was delivered. The patient made an uncomplicated recovery and was discharged home on the second postnatal day. The peripartum potassium was kept within the normal range with intravenous as well as oral potassium supplementation. No arrhythmias were reported. CONCLUSION: Assessment of the patient at an early stage in her pregnancy allowed for a multidisciplinary approach to this patient and her medical problems. A plan was made to avoid known precipitating factors during labour, delivery and the postnatal period well in advance of her date of confinement, leading to a successful outcome for mother and child. PMID- 10674512 TI - Subcutaneous emphysema following trans-cricothyroid membrane injection of local anesthetic. AB - PURPOSE: To present a case of preoperative subcutaneous emphysema (SCE) as a complication of trans-cricothyroid membrane (TCM) injection of lidocaine for awake intubation. CLINICAL FEATURES: A 48-yr-old man with cervical myelopathy was scheduled for elective cervical discectomy. Airway topical anesthesia consisted of lidocaine pledgets and TCM injection. After successful awake fibreoptic intubation was performed, SCE was noted in the neck region. The main differential diagnosis of preoperative SCE included air leak via the anterior needle track from TCM injection or disruption of mucosal membrane in the aerodigestive tract. The latter was excluded by panendoscopy and an upper GI swallow study. The most likely explanation for SCE was air leak from the anterior needle tract. The subcutaneous emphysema resolved spontaneously without sequella. CONCLUSION: Subcutaneous emphysema is a rare but potentially serious complication of TCM injection of lidocaine. Anesthesiologists should be familiar with the differential diagnosis, investigations and management of SCE. PMID- 10674513 TI - Differential lung ventilation and emergency hyperbaric oxygenation for repair of a tracheal tear. AB - PURPOSE: To report the anaesthetic management of a case of tracheal rupture, using different types of ventilation and additional hyperbaric oxygenation (HBO). CLINICAL FEATURES: An 8 cm postintubation tracheal tear was repaired in a 66-yr old woman with acute myocardial reinfarction, mediastinal and subcutaneous emphysema, cardiac failure and unrecognized lymphoma. Intraoperative monitoring included dual oximetry: arterial (SaO2) and mixed venous saturations (SvO2). Maintenance of free surgical access and a series of life-threatening events like dislocation of the jet catheter required many ventilation modes. An episode of supraventricular tachycardia was interrupted by cardioversion. Differential lung ventilation with a combination of conventional and high-frequency jet ventilation (HFJV) modes preserved oxygenation (PO2 139.2 mm Hg, PCO2 42.4 mm Hg, FiO2 1.0) until acute tube obstruction and decrease of saturation values (SaO2 58%, SvO2 45%) required emergency HBO: immediate cardiac and respiratory stabilization was provided by double-lung HFJV and apneic oxygenation under hyperbaric conditions at 2.5 atmospheres absolute for 35 min (SaO2 100%). The patient recovered from surgery but died of non-Hodgkin lymphoma. CONCLUSION: The combination of different ventilation modes including HFJV and the additional use of HBO resulted in sufficient oxygenation during tracheal repair. PMID- 10674514 TI - Biting the laryngeal mask: an unusual cause of negative pressure pulmonary edema. AB - PURPOSE: To describe negative pressure pulmonary edema due to biting of the laryngeal mask tube at emergence from general anesthesia. CLINICAL FEATURES: A healthy patient underwent general anesthesia using a laryngeal mask airway and mechanical ventilation. During recovery, the patient strongly bit the laryngeal mask and made very forceful inspiratory efforts until the mask was removed. Five minutes later, the patient developed dyspnea and had an hemoptysis of 50 ml fresh blood. Chest radiograph showed bilateral alveolar infiltrates. Pharyngo-laryngeal examination was normal. Bronchoscopy revealed no injury but diffuse pink frothy edema fluid. Clinical examination and chest radiograph became normal after 12 hr of nasal oxygen therapy confirming airway obstruction as the most available cause of this pulmonary edema. CONCLUSION: Airway obstruction due to biting of a laryngeal mask tube may result in negative pressure pulmonary edema. PMID- 10674515 TI - Analysis of anesthesia physician resources: projected Ontario deficit in 2005. AB - PURPOSE: To clarify the recent perception of shortfalls in anesthesia physician resources, two models were used to assess these resources in Ontario, Canada. METHODS: Two models, demand-based and benchmarking, were used. In the demand based model estimated future supply and attrition were obtained from information on Ontario Ministry of Health funded trainees. Data from the Canadian Residents Matching Service and the Association of Canadian University Departments of Anesthesia were also used. Current demand was identified from a telephone survey of Departments of Anesthesia in ten Ontario cities. The number of anesthesia practitioners in Ontario was estimated from the 1996 Canadian Anesthesiologists' Society Physician Resource Database (CASPRD) in the demand-based model. In the benchmarking model, using Alberta as the closest published analogue to Ontario, the annual specialist growth rate in Ontario since 1986 was calculated in the literature as 2.8%/yr for 1986-1994. The number of anesthesiologists in Ontario from the 1986 CASPRD was used to calculate need based on that growth rate. Results are compared with population to anesthesiologist (P/A) ratios calculated from Statistics Canada population data and physician numbers from CASPRD. RESULTS: A shortfall in the number of anesthesiologists has been identified. The P/A ratio worsened by 17.6% from 1986 to 1996. The demand-based model indicated that the shortfall is increased from a current deficit of 40 to 68 by 2005, using CASPRD. Benchmarking showed that the estimated shortfall in 1994 was 131. CONCLUSION: This conservative approach indicates that the shortfall in anesthesiologist physician resources will worsen by 2005. PMID- 10674516 TI - Cervical plexus anesthesia for carotid endarterectomy: comparison of ropivacaine and mepivacaine. AB - PURPOSE: To evaluate the effectiveness of cervical plexus block performed with ropivacaine 0.75% or 1%, or mepivacaine 2%. METHODS: In a prospective, randomized, double-blind study, 60 patients received deep cervical plexus block with 0.2 ml x kg(-1) divided among C2-C4 injections using ropivacaine 0.75% and 1% or mepivacaine 2%. A blinded observer recorded loss of pin-prick sensation every minute in the C2-C4 dermatomes until readiness for surgery. Then, a superficial cervical block was performed with 0.15 ml x kg(-1) lidocaine 1%. The need for intraoperative supplemental analgesia and degree of pain and time of first postoperative pain medication were also recorded. RESULTS: General anesthesia was not required to complete surgery in any case. No differences in the need for intraoperative supplemental analgesia was observed (7, 6, and 9 patients with ropivacaine 0.75% and 1% or mepivacaine 2%, respectively). Readiness to surgery required 15 (10-25) min with ropivacaine 0.75%, 18 (8-20) min with ropivacaine 1%, and 15 (5-20) min with mepivacaine 2% (P = NS); while patients asked for first postoperative pain medication after 10 (4-13) hr and 9 (6.5 - 11) hr with ropivacaine 0.75% and 1% compared with 5 (0-8) hr with mepivacaine 2% (P<0.05). CONCLUSION: Ropivacaine 0.75% or 1% are appropriate choices when performing cervical plexus anesthesia for carotid endarterectomy, providing nerve block characteristics similar to those of mepivacaine 2%, but with the advantage of longer postoperative pain relief. PMID- 10674517 TI - Paroxystic systemic hypertension during inhalation induction with sevoflurane 8%. PMID- 10674518 TI - Transthoracic echocardiography pre-, intra-, and postoperatively. PMID- 10674519 TI - Axillary blockade by the targeted method. Added benefit? PMID- 10674520 TI - Sevoflurane sedation. PMID- 10674521 TI - Stroke. PMID- 10674522 TI - Delta CK-MB outperforms delta troponin I at 2 hours during the ED rule out of acute myocardial infarction. AB - It has been shown that a rise in creatine kinase MB bank (CK-MB) of > or = + 1.6 ng/mL in 2 hours is more sensitive and equally specific for detection of acute myocardial infarction (AMI) as compared with a 2-hour CK-MB > or = 6 ng/mL during the emergency department (ED) evaluation of chest pain. Because cardiac specific troponin I (cTnI) is thought to have similar early release kinetics as compared with CK-MB mass, we undertook a retrospective cohort study in 578 chest pain patients whose baseline CK-MB and cTnI was less than two times the hospital's upper limits of normal and who underwent a 2-hour CK-MB and cTnI to compare sensitivities and specificities of the 2-hour delta CK-MB (deltaCK-MB) and delta cTnI (delta cTnI) for AMI and 30-day Adverse Outcome (AO). Thirty day AO was defined as AMI, life-threatening complication, death, or percutaneous transluminal coronary angioplasty (PTCA)/coronary artery bypass graft (CABG) within 30 days of ED presentation. Optimum delta values were determined by choosing the smallest cutoff value greater than the assay precision where the deltaCK-MB and delta cTnI had a positive likelihood ratio for 30-day AO of > or = 15. A deltaCK-MB > or = +1.5 ng/mL was more sensitive than a deltaTnI > or = +0.2 ng/mL for AMI (87.7% versus 61.4%; P < .0005) and 30-day AO (56.7% versus 42.3%; P < .005). There were no differences in specificities for AMI and 30-day AO. Combining the two tests (MBdelta > or = +1.5 ng/mL and/or a deltaTnI > or = +0.2 ng/mL) resulted in an incremental increase in sensitivity of 89.5% for AMI and 61.9% for AO (P < .005). Patients with either a rise in CK-MB of > or = +1.5 ng/mL or rise in cTnI of > or = +0.2 ng/mL in 2 hours should receive consideration for aggressive antiischemic therapy and further diagnostic testing before making an exclusionary diagnosis of nonischemic chest pain. PMID- 10674523 TI - Predicting discharge in uncomplicated near-drowning. AB - To determine if routine, noninvasive parameters could be measured which predict early (4-6 hour) discharge from the emergency department (ED) in mildly symptomatic and asymptomatic victims of childhood near-drowning, a retrospective cohort study was undertaken. Patients with fresh water near-drowning were studied over a 3-year period who presented with Glascow Come Scale (GCS) > or =13 and required no advanced life support prior to or < or =4 hours after ED presentation. Three groups of patients were found: 39 patients (81%) had normal pulmonary examination (PEx) and normal room air oxygen saturation (RASaO2) by 4 to 6 hours and did not deteriorate during the hospital admission (<24 hours); 5 patients (10%) had normal PEx by 4 to 6 hours and RASaO2 by 8 to 12 hours and did not deteriorate during hospitalization (<24 hours). Four patients (8%) were hospitalized for more than 24 hours. No patient with normal RASaO2 at 6 hours deteriorated while in the hospital (CI 92.3-100%). Children who present to the ED with GCS > or =13 and have normal PEx/respiratory effort and RA-SaO2 more than 95% at 4 to 6 hours after ED presentation can be safely discharged home. PMID- 10674524 TI - Use of the intubating laryngeal mask airway by medical and nonmedical personnel. AB - The intubating laryngeal mask airway (ILMA) is a newly available device designed to allow for blind endotracheal intubation and treatment of patients with difficult airways. We studied the intubation success rates and speed with initial use of this device on an intubation manikin to determine whether this device might be easily used by trained and untrained personnel. Rapid and successful intubation with a device requiring limited or no training could have widespread implications for both health care providers and laypersons. The study consisted of 2 parts. In part 1, health care providers with intubation experience, health care providers without prior intubation experience, and nonmedical personnel were instructed to enter a room and intubate a manikin using the ILMA. A single page set of schematic directions was provided within the ILMA setup. The main outcomes were the intubation success rate and the time required for successful ventilation and intubation. In part 2, participants were retested after a standardized <60 second device demonstration. The 111 participants in the study included 44 emergency physicians (40%), 21 anesthesiologists (19%), and 46 other medical or nonmedical personnel (41%). On first attempted use of the device, and with no prior training, 59% of all participants successfully intubated the manikin. Attending and resident physicians had an 83% initial success rate. The median time to ventilation was 47 seconds, and the median time from ventilation until intubation was 29 seconds. Following the <60 second demonstration, 108 of 111 (97%) participants achieved success, with the median time to ventilation 18 seconds, and the median time from ventilation until intubation 17 seconds. All attending and resident physicians succeeded in intubation following the demonstration. Success rates on first attempt correlated with level of training, prior intubation experience, and prior LMA use (all P < .001). After a <60 second demonstration, medical and nonmedical personnel with and without prior intubation training can successfully use the ILMA to rapidly establish an airway in a manikin model. The ILMA should be further studied to determine if it may permit endotracheal intubation by first responders, paramedical personnel, and other medical staff with limited or no laryngoscopy skills. PMID- 10674525 TI - Health promotion practices of emergency physicians. AB - In this article we describe health promotion practices of emergency physicians (EPs). A survey was mailed to members of the West Virginia American College of Emergency Physicians. Main outcomes included the EP's beliefs regarding health promotion, perceived roles in health promotion, and perceived effectiveness in modifying the behavior of patients. Over 90% of respondents routinely asked about cigarette smoking and half about alcohol use. A minority routinely asked about illicit drug use, diet, exercise, domestic violence, or stress. The majority stated they were the main person responsible for patient health education in their emergency department (ED). Most felt prepared to counsel patients about smoking (68%) and alcohol (59%), although very few described themselves as successful in helping patients change their behavior. Although EPs feel responsible for promoting the health of their patients, only a minority reported routinely screening and counseling patients about prevention and most were not confident in their ability to help patients change their health-related behaviors. PMID- 10674526 TI - Phospholipase A2-induced coagulation abnormalities after bee sting. AB - We will examine the correlation between various bee venom phospholipase A2 (PLA2) concentrations and several parameters of coagulation in human plasma in order to offer a rationale for requesting a particular laboratory coagulation test after bee sting(s). We will also evaluate in vitro the influence of clinically available drugs with a noncompetitive inhibitory effect on PLA2 on the anticoagulant effect of bee venom PLA2. Prothrombin index (PTi), partial thromboplastin time (PTT), antithrombin III (AT III), soluble fibrin monomers (SFM), the activity of coagulation factors I, II, V, and VIII, and thrombelastography (TEG) parameters (split point [Sp], reaction time [R], kinetic time [K], coagulation time [R + K], maximal amplitude [MA], and the growth angle [alpha]) were determined before and after addition of 1.4, 2.7, and 4.1 units (1, 2, and 3 microg protein respectively) of bee venom PLA2. Linear regression was used to determine the significance of the relationship between these coagulation parameters and bee venom PLA2 concentrations used. To study the influence of ketamine, lidocaine, magnesium, furosemide, and cromolyn on the anticoagulant effect of bee venom PLA2, PTi and factor II- and V-activities were measured before and after addition of 2.7 units of PLA2 and PLA2 plus one of the tested substances. Determinations of F II, PTi, F V, and F VIII showed a negative correlation to bee venom PLA2 concentration (r = -0.88, -0.86, -0.81, and -0.79 respectively). A positive correlation was found for PTT (r = 0.69). FII- activity and PTi correlated better with bee venom PLA2 concentration than other parameters. F I, AT III, and SFM showed no changes. Whereas Sp, R, and K were prolonged by bee venom PLA2 and a was reduced, there was no correlation to the PLA2 concentration. Addition of none of the 5 substances could correct the effects of bee venom PLA2 on the coagulation. In a patient with toxic reaction or a severe anaphylactic reaction after bee sting(s) we suggest determinations of FII and/or PTi. This will allow a quick and economical assessment of coagulation abnormalities after bee sting(s). Noncompetitive PLA2-inhibitors (ketamine, lidocaine, magnesium, furosemide, and cromolyn) are unable to correct in vitro the anticoagulant effect of bee venom PLA2. They cannot be recommended at this stage for this purpose. Further investigations with competitive PLA2-inhibitors are warranted. PMID- 10674527 TI - Duration of patient immobilization in the ED. AB - In this article we seek to determine the duration of immobilization in patients presenting to the emergency department (ED). We conducted a 10-week prospective study of a convenience sample of patients transported to a level one trauma center immobilized with a backboard and cervical collar. Total backboard time (TBT) was measured from the time the ambulance left the scene to the time the patient was removed from the backboard, while total ED backboard time (TEDBT) was measured from the time of arrival at the ED to the time of backboard removal. There were 138 patients entered in the study. Insufficient data excluded 36 patients from further analysis. TBT was available for 92 patients and averaged 63.63 (+/-45.87) minutes. Dividing patients into those who were removed from the backboard prior to radiographs (n = 85), the TBT average was 53.9 minutes (+/ 30.1), whereas the average for those who had radiographs prior to removal from the backboard (n = 7) was 181.3 minutes (+/-41.6). There were 102 patients for whom TEDBT was available and averaged 46.36 (+/-44.88) minutes. Dividing patients into those who were removed from the backboard prior to radiographs (n = 95), the TEDBT average was 37.6 minutes (+/-29.6), whereas the average for those who had radiographs prior to removal from the backboard (n = 7) was 165.3 minutes (+/ 49.7). Patients are left on backboards for significant periods of time even when no radiographs are taken prior to backboard removal. PMID- 10674528 TI - Global brain ischemia produced by clamping left subclavian artery and bicarotid trunk in the rabbit. AB - Thirty-five rabbits were divided randomly into 5 groups: sham operation, 10 minutes clamping bicarotid trunk (partial ischemia, PI), and 3 groups of 5, 7, and 10 minutes clamping left subclavian artery and bicarotid trunk (global ischemia, GI). Systolic arterial pressure increased slightly in the PI group, but doubled in the GI groups during clamping. Heart rate did not change in the PI group, but decreased transiently in the GI groups during clamping. Brain temperature decreased gradually in the GI groups during clamping, but did not change in the PI group. Necrotic changes were present 96 hours later in approximately 50% of the hippocampal CA1 cells in the GI groups, but in none of the cells in the PI and sham operation groups. The present results may indicate that clamping left subclavian artery and bicarotid trunk in the rabbit brings about global brain ischemia. PMID- 10674529 TI - The efficacy of oral deferiprone in acute iron poisoning. AB - Due to its high cost and need for parenteral administration, the standard iron chelator deferoxamine is not used in many individuals with acute and chronic iron poisoning worldwide. Deferiprone is the first oral iron chelator to be shown to be effective in chronically iron overloaded thalassemia patients. Its efficacy, by oral administration, in acute iron poisoning has not been tested. Our objective was to determine whether orally administered deferiprone can reduce the mortality of rats following acute, toxic, oral doses of iron. Rats were administered 612 mg/kg elemental iron orally, corresponding to LD50 in the species tested. Two other groups received the same oral dose of iron followed by oral deferiprone: 800 mg/kg and 800 mg/kg, followed by another dose of 800 mg/kg 2 hours later. Coadministration of 800 mg/kg deferiprone with the iron decreased mortality from 30% to 6.6% after 2 hours (P = .02), from 40% to 16.6% after 12 hours (P = .04), and from 53.3% to 20% after 24 hours (P = 0.007). Mortality was also significantly decreased among animals coadministrated 2 repeated doses of deferiprone of 800 mg/kg with iron, to 0%, 9%, and 18%, and 2, 12, and 24 hours postdrug administration, respectively (P = .04, .05, .04, respectively). Histologically, there was a dose-dependent decrease in iron accumulation in the gastrointestinal tract. Orally administered deferiprone can decrease morbidity and mortality caused by acute iron overdose in rats. Oral deferiprone holds promise in the treatment of iron poisoning in humans. PMID- 10674530 TI - A preliminary evaluation of emergency ultrasound in the setting of an emergency medicine training program. AB - In this article we seek to evaluate the diagnostic accuracy of emergency physicians performing emergency ultrasonography in the setting of an emergency medicine training program. A prospective observational study was performed at an inner city Level I trauma center with an emergency medicine residency training program. From July 1994 to December 1996 a convenience sample of ultrasound exams was recorded. The diagnostic quality ("acceptable or technically limited") was determined by a board-certified cardiologist or radiologist with fellowship training in ultrasonography. The emergency department interpretations were then compared to those of the blinded cardiologist or radiologist. Four hundred and fifty-six ultrasound examinations were videotaped and entered into the study; 408 (89%) of the studies performed were determined to be "acceptable." The diagnostic accuracy (sensitivity, specificity, positive and negative predictive values) of these studies were as follows: cardiac, to rule out effusion (n = 67; 0.83, 0.98, 0.88, 0.98); transabdominal, to rule out abdominal aortic aneurysms (AAA), cholelithiasis, or free peritoneal fluid (n = 263; 0.91, 0.89, 0.88, 0.92); renal, to rule out hydronephrosis (n = 45; 0.94, 0.96, 0.94, 0.96); pelvic, to rule in intrauterine pregnancy (n = 33; 1.0, 0.90, 0.96, 1.0). The 48 "technically limited studies" included: 39 transabdominal (33 gallbladder, 1 abdominal aortic aneurysm, 5 free peritoneal fluid), 6 cardiac, 2 renal, and 1 pelvic ultrasound. This study suggests that emergency physicians with a minimal amount of training display acceptable technical skill and interpretive acumen in their approach to emergency ultrasonography. PMID- 10674531 TI - Diagnosis of acute thoracic aortic dissection in the emergency department. AB - In this article we try to determine how frequently emergency physicians (EPs) suspected the diagnosis in acute aortic dissection (AD). In this retrospective descriptive study, we identified all patients with the final diagnosis of AD initially evaluated in 1 of 3 emergency departments (EDs) over a 5-year period. Patients were included if AD was not suspected before ED evaluation. Patients undergoing thoracic aorta imaging as the initial ED study were defined as suspected AD. Forty-three patients totaling 44 presentations were identified. EPs suspected AD in 19 of 44 presentations. EPs suspected AD in 12 of 14 (86%) cases of chest and back pain and in 5 of 11 (45%) of chest pain. Thirteen of 39 (33%) painful presentations involved abdominal pain; EPs suspected AD in 1 of 13 (8%). EPs suspected the diagnosis in 43% of acute AD; location of pain was most predictive of a suspected diagnosis. PMID- 10674532 TI - Use of complementary and alternative medicine among Dominican emergency department patients. AB - This small, pilot study examined presenting complaint, brief health history, use of complementary and alternative medicine (CAM), and sociodemographic characteristics, among patients attending the emergency department (ED) of a large urban hospital. The sample (n = 50) was primarily Dominican and of low socioeconomic status. Almost half had used CAM for their presenting complaint or another health problem during the past year, most commonly in the form of medicinal plants made into herbal teas. CAM users were more likely to be female, longer-term residents of the United States, and to have also used religious practices for health problems. Subjects who had used CAM for any problem other than the presenting complaint during the past year rated its effectiveness higher than subjects who had used CAM for their presenting complaint. In conclusion, it is likely that a significant proportion of Dominican ED patients use CAM, suggesting that they should be asked about their CAM use during triage. PMID- 10674533 TI - Unstable cervical spine without spinal cord injury in penetrating neck trauma. AB - Cervical spine instability in the neurologically intact patient following penetrating neck trauma has been considered rare or non-existent. We present a case of a woman with an unstable C5 fracture without spinal cord injury after a gunshot wound to the neck. Considerations regarding the risk of cervical spine instability are discussed, as well as suggestions for a prudent approach to such patients. PMID- 10674534 TI - Sternoclavicular joint injuries. AB - Injuries to the sternoclavicular (SC) joint are infrequently encountered. However, retrosternal SC joint dislocations are potentially life-threatening injuries which must be recognized by the examining physician and treated as soon as possible. Plain radiography often fails to fully distinguish SC joint injuries, and computed tomography has emerged as the diagnostic modality of choice for defining the injury complex and surrounding injuries. We have encountered 6 cases of SC joint injuries over the past 3 years and describe their presentation and management. PMID- 10674535 TI - The mosh pit experience: emergency medical care for concert injuries. AB - Effective planning is essential for medical personnel preparing to provide emergency care at mass gatherings. At large concerts where audience members participate in "moshing," crowd surfing, and stage diving, there may be a potential for a dramatic increase in injuries requiring medical attention. Injuries seen at emergency medical stations at 3 concerts, all with large mosh pits, over 4 event days were recorded and evaluated. Each event day had over 60,000 attendees. A total of 1,542 medical incidents (82.9 per 10,000) were reported over the 4 event days. There were 37% (466 patients, 25.1 per 10,000) of incidents related to moshing activity. Hospital transport was required for 2.5% (39 patients, 2.1 per 10,000) of medical visits with 74% (29 patients, 1.5 per 10,000) of those transported being for mosh pit-related injuries. When planning emergency medical care for such concerts with mosh pits, the potential for an increase in the number of medical incidents and injuries requiring medical attention and hospital transport should be taken into account for efficient medical coverage. PMID- 10674536 TI - Green lynx spider (Peucetia viridans) envenomation. AB - Four cases of envenomation by green lynx spiders (Peucetia viridans) are reported. Despite the unusual appearance and occasional aggressive behavior of this spider, envenomation caused only local pain, pruritus, erythema, and induration. No local necrosis or systemic symptoms occurred. Treatment included tetanus immunization, wound care, and symptomatic therapy. PMID- 10674537 TI - Benefits of an informational videotape for emergency department patients. AB - To determine if an informational videotape affected patient's attitudes towards their emergency department visit, we conducted a prospective study using a convenience sample of patients waiting to be seen at a southern California emergency department. Patients waiting to be treated were randomized to view an informational videotape or to receive standard management (no videotape). The informational videotape lasted 6 minutes and served to orient the patients to the emergency department. It showed the sequence of steps from entry into the department to discharge, the nature of triage and causes for delays, the different services offered by the emergency department, and the roles of each of the department staff members. One week after discharge, patients were contacted at home by telephone and were asked to rate various aspects of their emergency department experience. A comparison of the telephone survey rankings between those who viewed the videotape (98 patients) and those who did not (100 patients) revealed statistically significant improvements in the former group on questions about level of anxiety and appropriateness of delays. An informational videotape for patients in waiting areas may be a useful tool to educate about emergency medical services, to reduce anxiety, and to improve satisfaction with the emergency department stay. PMID- 10674538 TI - Ultrasound guided reduction of pediatric forearm fractures in the ED. AB - Reducing badly displaced or angulated pediatric forearm fractures in the emergency department can be difficult. Multiple attempts at reduction may be required, with repeated trips to the radiology department, before an adequate reduction is achieved. We have recently found that bedside ultrasound by emergency physicians is very helpful in guiding the reduction of difficult forearm fractures, allowing the physician to assess the adequacy of the reduction at the patient's bedside. In this report, we describe the technique we have developed for ultrasound-guided fracture reduction and present three case histories showing the usefulness of this technique. PMID- 10674539 TI - Delayed presentation of abdominal bleeding in a teenage boy after a fall. AB - The delayed presentation of an abdominal bleed in a victim of a fall is a rare occurrence. In the multiple injured patients, even with an intact sensorium, competing pain from associated injuries may mask the pain from a occult injury. Although a rare occurrence of abdominal injury in an asymptomatic neurologically intact patient, in the patient requiring a computed tomography scan of a spinal fracture, it may be worthwhile to image the abdomen and pelvis as well to rule out a concomitant occult abdominal injury. Current literature regarding injuries associated with falls from height are discussed that support this position and the delayed manifestation of an abdominal bleed in a 17-year-old boy 1 day after a fall is presented. PMID- 10674540 TI - A sighting of orbital pseudotumor. AB - A 39-year-old woman developed bilateral proptosis, photophobia, and pain with extraocular movements over the course of 5 days. Her findings initially were ocular pain and photophobia which progressed to periorbital edema and nasal discharge ultimately resulting in proptosis with vertical globe displacement and decreased visual acuity. She was diagnosed with corneal abrasion and sinusitis respectively during two initial emergency department visits. On her third visit to the emergency department within 4 days, she developed acute visual deficits. The patient was subsequently diagnosed with orbital pseudotumor after computed tomography scan revealed inflammation of orbital structures bilaterally. PMID- 10674541 TI - Cases in electrocardiography. PMID- 10674542 TI - Unruptured cerebral aneurysm producing a thunderclap headache. AB - A sudden and severe headache is the most common presentation of an acutely ruptured cerebral aneurysm. A similar headache in the absence of subarachnoid blood has rarely been ascribed to an unruptured cerebral aneurysm, but may result from acute aneurysm expansion and indicate a high risk of future rupture. We present a patient who developed a sudden, severe, "thunderclap" headache, with no associated neurological deficit. Computed tomogram and lumbar cerebral spinal fluid obtained 5.5 hours after headache onset were negative for subarachnoid hemorrhage. The patient underwent cerebral angiography which revealed a posterior communicating artery aneurysm with an associated daughter aneurysm. Craniotomy and clip obliteration of the aneurysm were performed. The aneurysm dome was very thin and there was no evidence of recent or old hemorrhage. A "thunderclap" headache without subarachnoid hemorrhage may be an important harbinger of a cerebral aneurysm with the potential for future rupture. Early recognition and neurovascular imaging of aneurysms presenting in this rare fashion are warranted. PMID- 10674543 TI - Continuous and bilevel positive airway pressure in the treatment of acute cardiogenic pulmonary edema. AB - Patients with acute cardiogenic pulmonary edema (ACPE) are commonly seen in the emergency department (ED). Although the majority of patients respond to conventional medical therapy, some patients require at least temporary ventilatory support. Traditionally, this has been accomplished via endotracheal intubation and mechanical ventilation, an approach that is associated with a small but significant rate of complications. The past 2 decades have witnessed increasing interest in methods of noninvasive ventilatory support (NVS), notably continuous positive airway pressure (CPAP) and bilevel positive airway pressure (BiPAP). We review the physiological consequences, clinical efficacy, and practical limitations of CPAP and BiPAP in the management of ACPE. PMID- 10674544 TI - Current controversies in the management of minor pediatric head injuries. AB - Each year hundreds of thousands of children receive care in emergency departments after head injury. Minor head injuries account for a majority of these injuries. The prevalence, morbidity, and costs associated with pediatric minor head injuries make it an important topic. We review the management of pediatric minor head injury, emphasizing current areas of controversy, including criteria for neuroimaging, indications for hospitalization, the role of anticonvulsant therapy, and the long-term neurobehavioral sequelae of pediatric minor head injury. PMID- 10674545 TI - Future of the emergency physician: subject or citizen? AB - Seventy-seven percent of emergency physicians (EPs) work as either employees or independent contractors (ICs). In contrast, other hospital-based physicians such as radiologists and anesthesiologists have a much higher percentage of ownership in their medical practices. The development of a high percentage of nonownership arrangements among EPs finds a useful historical comparison in the industrialization of nonhealth care workers over the past 100 years. Unless significant changes occur in emergency medicine (EM) organization and practice structures, EPs will have less self-determination over their practice compared with other specialties. This will inevitably result in less self-determination for their future. Combined with the great strides EM has achieved as a specialty, EPs' brightest future lies in being citizens of a broader, more expansive, all encompassing EM practice. PMID- 10674546 TI - The intravenous use of coconut water. AB - Medical resources routinely used for intravenous hydration and resuscitation of critically ill patients may be limited in remote regions of the world. When faced with these shortages, physicians have had to improvise with the available resources, or simply do without. We report the successful use of coconut water as a short-term intravenous hydration fluid for a Solomon Island patient, a laboratory analysis of the local coconuts, and a review of previously documented intravenous coconut use. PMID- 10674547 TI - A bartender fixes a curious drink. PMID- 10674548 TI - Coronary vasospasm in a patient suffering from sarin poisoning. PMID- 10674549 TI - Ruptured thoracic aorta aneurysm after spontaneous pneumothorax drainage. PMID- 10674550 TI - Intussusception of a normal appendix. PMID- 10674551 TI - Inferior hip dislocation in an adult: does a rare injury now have a common mechanism? PMID- 10674552 TI - Behcets disease in an African American man. PMID- 10674553 TI - The effect of introducing bedside TV sets on patient satisfaction in the ED. PMID- 10674554 TI - CO poisoning caused by inhalation of CH3Cl contained in personal defense spray. PMID- 10674555 TI - Fluid loading: neither safe nor efficacious in the treatment of the alcohol intoxicated patient in the ED. PMID- 10674556 TI - The results of a randomized controlled trial of hydrocortisone in acute exacerbation of COPD. PMID- 10674557 TI - The spectrum of anaphylaxis. PMID- 10674558 TI - Role of staphylococcal superantigens in atopic dermatitis: from colonization to inflammation. AB - OBJECTIVE: This review article has been prepared in order to enable the readers to understand the role of staphylococcal superantigens (SsAgs) in atopic dermatitis (AD). DATA SOURCES: MEDLINE literature search was performed for obtaining references. Recent reviews, research articles, poster presentations, and letters (to the editor) were meticulously reviewed. RESULTS: (1) SsAgs contribute to the pathogenesis of cutaneous inflammation in AD with five potential mechanisms: Direct stimulation of antigen presenting cells (APCs) and keratinocytes. Stimulation of T cell proliferation [superantigenic binding to T cell receptor (TCR)]. Expansion of skin-homing cutaneous lymphocyte antigen (CLA) (+) T cells. The role of superantigens as allergens. Reduction of apoptosis. (2) Effectiveness of antibiotic therapy in AD patients without signs of bacterial infection is still under discussion. If signs of skin infection are present, antibiotic therapy (topical/oral) may help exacerbations of AD. Prolonged topical/oral antibiotic therapy, however, may cause development of antibiotic resistant strains of Staphylococcus aureus (SA). CONCLUSIONS: Atopic dermatitis is a genetically determined, chronically relapsing, inflammatory skin disease which has many aspects and a complex immunopathogenesis involving both immediate and cellular immune responses. While the pathogenic role of SsAgs may not be of primary importance, SsAgs appear to be one of the important triggering factors that contribute to the cutaneous inflammation in AD. We suggest that staphylococcal colonization does not always mean SsAg-mediated inflammation, and anti-staphylococcal treatment should be considered in cases with signs of bacterial infection. PMID- 10674559 TI - Two contrasting cases of anaphylaxis seen simultaneously. PMID- 10674560 TI - Comparison of the Burkard and Allergenco MK-3 volumetric collectors. AB - BACKGROUND: The Burkard sampler is a widely used volumetric pollen and spore collector, in part, because it is wind-oriented, it has consistent flow characteristics, and it permits time-discrimination of collected particles. Its main disadvantages are that it is heavy, expensive, and visual counting is very time-consuming. A less-expensive volumetric collector with time discrimination capabilities could permit more widespread particle counting which would enhance our understanding of aerobiology. OBJECTIVE: The objective of this study is to compare the collection recoveries of the Burkard sampler with a less-expensive non-wind oriented collector, the Allergenco MK-3, under various wind speeds. METHODS: Pollen and spore counts were compared on 20 sampling days during the spring pollen season using a Burkard and Allergenco MK-3 located next to each other on the roof of a 5-story hospital building. A weather station was placed nearby and wind velocity was concurrently measured. RESULTS: The median wind velocity was 6 miles/hour with a maximum of 35 miles/hour. The Burkard and Allergenco MK-3 collectors displayed similar collection characteristics at all wind velocities for both pollen and spores. The Burkard gave lower counts than the Allergenco when absolute particle counts were low and similar values at higher absolute counts. CONCLUSIONS: Given our data, we conclude that ambient wind speed has no significant effect on collection efficiency at velocities commonly found on the roof of our hospital and that the collection characteristics of the Burkard and Allergenco MK-3 are comparable. PMID- 10674561 TI - Distribution of primary immunodeficiency diseases diagnosed in a pediatric tertiary hospital. AB - BACKGROUND: Advances in immunologic techniques in recent years have led to increased recognition of primary immunodeficiency disorders, with IgA deficiency the most common phenotype reported by most registries. There have also been reports of increased associated incidence of autoimmunity, allergy, and other diseases. OBJECTIVES: We wished to determine the percentage of different primary immunodeficiency disorders seen in a pediatric tertiary hospital and to determine the association of primary immunodeficiency disorders with other diseases that are not part of classic immunodeficiency disorders. METHODS: We performed a retrospective review of the patients referred to our allergy/immunology clinic for immunologic evaluation of recurrent infections during an 8-year period. We also reviewed pathology reports with postmortem diagnosis of immunodeficiencies not identified while patients were alive. RESULTS: Of the 91 patients with primary immunodeficiency disorders evaluated, the majority had predominantly antibody deficiencies (67%). The most common phenotype was specific antibody deficiency with normal immunoglobulins (23.1%), defined as inability to mount an adequate response to pneumococcal polysaccharides followed by IgG2 subclass deficiency (17.6%). These two phenotypes were diagnosed mostly in the last 2 years of the survey. Associated diseases, found in 40% of patients, were mostly allergic conditions followed by syndromic/chromosomal disorders. CONCLUSION: The study reveals that specific antibody deficiency with normal immunoglobulins followed by IgG2 subclass deficiency was the most frequently diagnosed primary immunodeficiency disorder in our patient population. It also indicates that immunodeficiency disorders should be considered in patients with other abnormalities like allergic and syndromic/chromosomal disorders that present with recurrent infections. PMID- 10674562 TI - Carpet properties that affect the retention of cat allergen. AB - BACKGROUND: Although the importance of carpeting on airborne levels of cat allergen (Fel d 1) has been demonstrated, no studies have been performed to determine specific properties of carpet that may affect its retention and removal. OBJECTIVES: This study characterizes factors that affect the retention of cat allergen on tufted carpets. The experiments were designed to test the hypothesis that the amount of allergen-containing dust recovered from vacuum samples of tufted carpet sources was dependent on micro (fiber) or macro (construction) retention properties of carpets. METHODS: Twenty-six types of custom manufactured carpet were spiked and embedded with reference dust containing Fel d 1. A standardized vacuum surface sampler was used to recover dust from the samples. Allergen was assayed using a standard, monoclonal antibody ELISA. RESULTS: Carpet-surface area and fluorocarbon-fiber treatments were found to have the largest effects on retention and recovery of cat allergen. The style per se of a carpet, such as loop or cut pile, does not affect allergen retention. These results are generally in agreement with previous studies on dust mite allergen retention. CONCLUSIONS: Carpets that are easiest to clean would have the following properties for release of cat allergen and in this order: low pile density and height, fluorocarbon coating of fibers, high denier per filament, and a fiber shape with a low surface area. PMID- 10674563 TI - Cord blood IgE: its determinants and prediction of development of asthma and other allergic disorders at 12 months. AB - BACKGROUND: The value of cord blood IgE in predicting the development of asthma and other IgE-mediated allergic diseases is unclear. OBJECTIVE: The purpose of this study is twofold: (1) to determine factors affecting cord blood IgE level and (2) to determine whether cord blood IgE predicts the development of asthma and other IgE-mediated allergic diseases in high risk (defined as those with at least one first degree relative with asthma or 2 first degree relatives with other IgE-mediated allergic diseases) infants at 12 months. METHODS: The study utilized cord blood obtained from a group of high risk infants who took part in a randomized controlled trial to assess the effectiveness of an intervention program in the primary prevention of asthma and other IgE-mediated allergic diseases. Total IgE and cotinine in the cord blood were measured. Assessment of the infants was done at 12 months for these diseases. RESULTS: Sixty-four (17.8%) infants had detectable total IgE in cord blood >0.5 kU/L. The proportion of infants with elevated cord blood IgE was significantly higher among nonwhites, birth during winter months, and those with a maternal history of asthma. There was no correlation between cord blood IgE and cord blood cotinine level. Cord blood IgE was found to be a significant predictor for the development of urticaria due to food allergy but not for other outcomes. CONCLUSION: Both genetic and environmental risk factors play a role in determining the level of IgE in cord blood. Cord blood IgE was a significant risk factor for the development of urticaria due to food allergy at 12 months of life. As urticaria due to food allergy is a prodrome for anaphylaxis, measurement of IgE in cord blood may be indicated in infants at high risk for developing allergic diseases so that preventive measures can be applied. PMID- 10674564 TI - Oral iron cutaneous adverse reaction and successful desensitization. AB - BACKGROUND: The oral administering of iron preparations sometimes produces adverse gastrointestinal effects. In contrast, cutaneous reactions are extremely rare. OBJECTIVE: We report a patient with several episodes of generalized pruritus and erythematous maculopapular eruption after receiving oral compounds of iron and on whom desensitization with oral iron was attempted. METHODS: We studied a female with microcytic anemia due to gynecologic blood loss who presented several episodes of cutaneous eruption after receiving oral compounds of iron. Skin prick-test and two simple-blind, placebo-controlled oral challenges were performed with various iron compounds, and finally desensitization with oral iron was carried out. RESULTS: Skin prick-test and patch-test with iron preparations were negative. Two simple-blind, placebo-controlled oral challenges were performed and the patient began experiencing similar cutaneous symptoms. We started a slow desensitization protocol using increasing doses until the target amount of the drug was tolerated without adverse effects. The chronic administration of oral iron therapy once a day for 9 months sustained the desensitized state and the anemia disappeared. CONCLUSION: We present methods to effectively manage iron supply for a microcytic anemia patient with cutaneous reactions due to oral iron compounds, to avoid repeated transfusions, slow desensitization with oral iron was successfully attempted. PMID- 10674565 TI - Comparison of outdoor allergenic particles and allergen levels. AB - INTRODUCTION: Spore and pollen counts have been used traditionally to determine aeroallergen exposure. Using a liquid based collector and enzyme immunoassays, we have developed methods for measuring airborne allergen concentrations. In this work we test the hypothesis that airborne allergen concentrations are directly related to spore and pollen counts. METHODS: Test samplers used included a high volume cyclonic liquid impinger (SpinCon) and a standard spore trap (Burkard). Samples were collected on a weekly basis from May to October and were analyzed microscopically for spores and pollen grains. The liquid samples were analyzed by enzyme-linked immunoassay for the presence of allergens from Alternaria, Cladosporium, Aspergillus, oak, fescue, ragweed, and plantain. Specific Alternaria allergens Alt al and GP70 also were measured. RESULTS: Pollen counts for the SpinCon and Burkard collectors were similar, though spore counts were lower with the SpinCon. Detectable amounts of three of the seven allergenic species including fescue, ragweed, and Alternaria were present in air samples. Concentrations of pollens were seen in their respective seasons while fungal allergen levels varied throughout the period. Allergen levels generally agreed with particle counts, however peak allergen levels and peak particle counts for individual species did not correlate well. CONCLUSIONS: At flow rates of 236 L/min, the SpinCon is comparable to the Burkard for counting airborne pollen and spores. Samples collected by the SpinCon permit quantitative determination of allergen levels in outdoor air. The poor correlation between measured airborne allergen and related particles indicates the potential for significant allergen exposure in the absence of identifiable particles in air. PMID- 10674566 TI - A cost-benefit analysis using a willingness-to-pay questionnaire of intranasal budesonide for seasonal allergic rhinitis. Rhinocort Study Group. AB - BACKGROUND: The cost-benefit of intranasal steroids for the treatment of seasonal allergic rhinitis is unknown. OBJECTIVE: To determine the cost-benefit of intranasal budesonide for seasonal allergic rhinitis. METHODS: Subjects who were symptomatic for a baseline period of 7 to 10 days were randomized to receive intranasal budesonide by Turbuhaler (400 microg) (n = 121) or aqueous spray (256 microg) (n = 121) once daily for 4 weeks. A willingness-to-pay questionnaire that measured benefits of treatment was administered before and at study completion. Costs were collected and compared with benefits. RESULTS: Subjects were willing to spend on average $15.89/wk (range $1 to $75) to alleviate the problems of seasonal ragweed rhinitis. Eighty percent of subjects felt that, with treatment, rhinitis had less of an impact on their lives, compared with previous years. The mean willingness-to-pay for the drug used during another ragweed season was $12.95/wk. This was 92% (95% CI, 85% to 100%) of the pre-treatment estimate. There was no relationship between an indirect assessment of income and willingness-to-pay estimates. The benefit was greater than the cost by a mean of $5.80/wk (95% CI, $3.52 to 8.08), P < .0001. There was no difference in costs, willingness-to-pay, or cost-benefit comparing delivery modes. A sensitivity analysis revealed the conclusions were robust. CONCLUSIONS: Intranasal budesonide is cost-beneficial in the treatment of seasonal allergic rhinitis and a willingness-to-pay questionnaire may provide a useful method to assess a therapy's benefit. PMID- 10674567 TI - Sparfloxacin for the treatment of acute bacterial maxillary sinusitis documented by sinus puncture. AB - PURPOSE: To evaluate the efficacy and safety of sparfloxacin in the treatment of patients with acute bacterial maxillary sinusitis, the microbiologic etiology of which was determined by maxillary sinus puncture. PATIENTS AND METHODS: Two hundred fifty-three patients enrolled in the open, noncomparative trial received sparfloxacin as a 400-mg loading dose followed by 200 mg once daily doses for a total of 10 days. One hundred ninety-eight patients were clinically evaluable and 82 were also bacteriologically evaluable. All treated patients were included in the safety analysis. Overall success was determined based on clinical success (resolution or reduction of signs and symptoms and sinus x-rays) and bacteriologic success (eradication and presumed eradication of baseline pathogens obtained by maxillary sinus puncture and aspiration). RESULTS: Overall success in the bacteriologically evaluable population at test-of-cure was 91.5% [75/82; 95% confidence interval (85.4%, 97.5%)]. For all pathogens, the eradication rate was 93.2% (109/117 baseline pathogens); individual pathogen eradication rates were 88.9% (16/18) for S. pneumoniae (including those strains exhibiting decreased susceptibility to penicillin); and 100% for H. influenzae (17/17), S. aureus (14/14), and M. catarrhalis (11/11). The majority of adverse events were of mild or moderate severity; the most frequently related adverse events were photosensitivity reaction, headache, nausea, and diarrhea. CONCLUSION: Sparfloxacin had an overall success rate of 91.5% for patients in this study and was generally well tolerated in the treatment of acute bacterial maxillary sinusitis. PMID- 10674568 TI - Effects of sodium cromoglycate on cytokine production following antigen stimulation of a passively sensitized human lung model. AB - BACKGROUND: Interleukin-5 (IL-5) and tumor necrosis factor-alpha (TNF-alpha) play key roles in bronchial asthma. Sodium cromoglycate (DSCG) and dexamethasone (Dex) are used in the treatment of asthma as anti-inflammatory agents. OBJECTIVE: We investigated whether DSCG inhibited the expression of IL-5 and TNF-alpha mRNA and proteins from isolated human lungs, and compared these findings with those of Dex. METHODS: Human lung specimens were passively sensitized with sera from atopic patients, then preincubated in the presence of DSCG (10(-3), 10(-4), 10( 5) M) or Dex (10(-6) M) for 2 hours. The specimens were stimulated with Dermatophagoides antigen, then cultured for 48 hours. The supernatant was collected 1, 2, 4, 8, 24, and 48 hours to measure IL-5 and TNF-alpha by enzyme linked immunosorbent assay. mRNA expression was examined by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Tumor necrosis factor-alpha protein reached a peak level at 4 hours (156.57 +/- 18.29 pg/mL). Dex decreased TNF-alpha protein to 31.86 +/- 4.67 pg/mL (P < .001). There was also a decrease of TNF alpha protein to 107.43 +/- 14.25 pg/mL by 10(-4) MD SCG (P < .001). Antigenic stimulation also increased the release of IL-5 protein at 4 hours and the peak level was observed at 24 hours (150.29 +/- 19.12 pg/mL). Dex decreased IL-5 protein to 28.57 +/- 5.27 pg/mL (P < .0001), 10(-4) M DSCG also decreased to 111.57 +/- 15.28 pg/mL (P < .05). RT-PCR analysis showed persistence of IL-5 and TNF-alpha mRNA expression from 1 to 24 hour after antigen stimulation. Dex but not DSCG inhibited IL-5 and TNF-alpha mRNA levels. CONCLUSION: Our results showed that DSCG significantly inhibited IL-5 and TNF-alpha production by human lung specimens, suggesting that it acts as an anti-inflammatory agent. PMID- 10674569 TI - Increased perception of dyspnea by inhalation of short acting beta2 agonist in patients with asthma of varying severity. AB - BACKGROUND: Poor perception of dyspnea in asthma may lead to a delay in starting appropriate treatment which is probably one of the factors contributing to death from asthma. OBJECTIVE: This study was carried out to determine whether impaired perception of dyspnea in patients with asthma of varying severity can be corrected by inhalation of short acting beta2 agonist treatment. METHODS: We enrolled 20 patients with asthma of varying severity. Forced expiratory volume in one second (FEV1) was measured before and 10 minutes after two puffs of salbutamol administered by metered dose inhaler. Perception of dyspnea was scored on the Borg scale during breathing through an inspiratory muscle trainer. RESULTS: After inhalation of short acting beta2 agonist treatment, the baseline Borg score was decreased significantly from 2.20 +/- 0.32 to 1.80 +/- 0.31 (P < .01). The Borg score during breathing with the highest resistance, on the contrary, was increased significantly from 6.25 +/- 0.35 to 6.90 +/- 0.35 after inhalation of short acting beta2 agonist treatment (P < .01). Highest resistance induced score difference from the baseline value (highest resistance-load score) was increased significantly from 4.10 +/- 0.46 to 5.25 +/- 0.42 (P < .01). There was no relationship between the change of Borg score from baseline value at each resistive load and the % change of FEV1 after inhalation of short acting beta2 agonist treatment. CONCLUSION: These studies demonstrate that inhalation of short acting beta2 agonist treatment decrease dyspnea, but increase perception of dyspnea induced by a resistive load in patients with asthma, and the mechanism of the increased perception may not be related to the increased airflow rate. It may be due to some local or central effects of bronchodilator drug on perception of asthma. PMID- 10674570 TI - Hypersensitivity pneumonitis resulting from community exposure to Canada goose droppings: when an external environmental antigen becomes an indoor environmental antigen. AB - BACKGROUND: In the past, hypersensitivity pneumonitis has been attributed to occupational, agricultural, or home environmental exposure. OBJECTIVE: This report describes the first case of hypersensitivity pneumonitis due to community exposure to droppings from Canada geese migrating through a suburban environment. METHOD: Clinical and serologic information was used in making the diagnosis of hypersensitivity pneumonitis. RESULTS: Serologic analysis demonstrated precipitating antibodies against goose droppings and against an extract made from washings from a filter taken from the patient's office. These studies also showed that the antigens in the office filter were goose dropping antigens. CONCLUSION: Hypersensitivity pneumonitis can result from exposure to goose dropping antigens in the community that enter buildings through ventilation systems. This represents a new form of an old disease. PMID- 10674571 TI - Cross-reactivity among conifer pollens. AB - BACKGROUND: There are increasing reports of Cupressaceae pollinosis from various geographic areas. Cross-reactivity among a limited number of species within the Cupressaceae family has been suggested. Juniperus ashei (mountain cedar) is the leading cause of respiratory allergy in South Texas. OBJECTIVE: This study examines in vivo and in vitro cross-reactivity among 12 Cupressaceae species, one Taxodiaceae species, one Pinaceae species, and an angiosperm. METHODS: Cross reactivity among pollen extracts of mountain cedar (MC) and the other 14 trees was investigated by: (1) prick skin testing of each tree pollen extract in ten patients with MC pollinosis. (2) Ouchterlony gel immunodiffusion employing rabbit antisera to MC. (3) IgE immunoblotting using high-titer MC pooled human sera, and immunoblot inhibition after pre-incubation with MC protein. (4) Monoclonal antibody immunoblotting using a murine monoclonal antibody with strong affinity for the gp40 major allergen of MC. RESULTS: Positive skin wheal-and-flare responses occurred to all 12 Cupressaceae and Japanese cedar (the Taxodiaceae), but not to the Pinaceae or the angiosperm. In Ouchterlony gels, lines of identity or partial identity formed between MC and all pollens except the Pinaceae and the angiosperm. Immunoblots demonstrated IgE binding to the 40 to 42 kD protein in each Cupressaceae, and to a parallel band in Japanese cedar at 43 to 46 kD. Immunoblot inhibition by MC pollen was complete for all trees. The monoclonal bound both the 40 to 42 kD protein in 11 of 12 Cupressaceae species and the 46 kD band in Japanese cedar, but bound no protein bands in the Pinaceae or the angiosperm. CONCLUSION: Pollen proteins of the 12 Cupressaceae (including MC) and the Taxodiaceae (Japanese cedar) are extensively cross-reactive. In particular, the MC major allergen, gp40, is cross-reactive with 40 to 42 kD proteins of the other Cupressaceae and with the Japanese cedar major allergen of 46 kD. Component based immunotherapy may someday allow a standard treatment for both Juniper allergic and C. japonica-allergic patients. PMID- 10674572 TI - Asthma inhaler use and barriers in a population-based sample of African-American and white adolescents. AB - BACKGROUND: There is little information on inhaler medication and barriers to use among a population-based sample of adolescents and whether possible variations in asthma treatment by ethnic group exist. OBJECTIVE: We describe the prevalence of inhaler use and identify barriers for proper use of asthma medication in a population-based sample of adolescents of which 34% are African-American. METHODS: A cross-sectional survey using the ISAAC (International Survey of Asthma and Allergies in Children) questionnaire was conducted in a school population based sample (n = 2056) of 13 to 14-year-old eight grade students in the Charlotte-Mecklenburg, North Carolina public school system. Questions were asked about symptom prevalence, asthma diagnosis, inhaler use, and barriers to care. RESULTS: Fourteen percent of the children (296/2056) reported using an inhaler in the last 12 months with no differences among African-American children and white children. Twenty-six percent of inhaler users were not allowed to carry their medication on their person while at school. Girls were more likely to be allowed to carry their inhalers at school and diagnosed asthmatic girls had a higher prevalence of wheezing in the last year (47%) compared with diagnosed asthmatic boys (35%). Smoking prevalence was higher in inhaler users (26%) compared to the study population (19%). CONCLUSIONS: Inhaler use is high in this population. Adolescents using inhalers need to reduce their smoking levels. Schools need to reevaluate their policies on the use of inhalers at school. PMID- 10674573 TI - Atopy is a risk factor for non-steroidal anti-inflammatory drug sensitivity. AB - BACKGROUND: There is scarce information in the literature about a possible association between atopy and certain clinical manifestations of NSAID sensitivity. OBJECTIVES: (1) To evaluate the prevalence of atopy in patients proved to be sensitive to cyclooxygenase inhibitors. (2) To assess cross reactivity to two alternative NSAIDs, paracetamol (acetaminophen) and nimesulide. METHODS: NSAID-sensitive patients attending an allergy clinic and unselected controls were prick tested with inhalant allergens. Oral challenges with NSAIDs were carried out by the single-blinded (SBOC) method. Clinical data about personal and family history of allergic and atopic diseases were obtained by a careful review of the medical records and by direct questioning by experienced allergists. RESULTS: Fifty patients had positive SBOCs to the suspected NSAID and only these were studied. A personal history of atopic diseases was present in 41 patients (82%) and 7 controls (14.5%), and a family history in 24 patients (48%) and 6 controls (12.5%). Prick skin tests with aeroallergens were positive in 39 of 45 patients tested (86.6%) and in 14 of 48 controls (29.1%), (P = .0001). Skin test positivity rates were higher in patients with cutaneous challenge reactions who responded to only one NSAID (single reactors) in comparison to cross-reactors (P = .04). The most frequent clinical manifestations of NSAID sensitivity were (1) cutaneous (angioedema, urticaria) in 34 patients, (2) blended (cutaneous plus respiratory) in 12, (3) respiratory in 3, and (4) anaphylactoid in 1. Aspirin, pyrazolone, paracetamol, and ibuprofen were the drugs more frequently implicated in these reactions. Cross-sensitivity with paracetamol and nimesulide were 32% and 25%, respectively. CONCLUSIONS: The prevalence of atopy is increased in challenge-proven NSAID-intolerant patients. The atopic condition may represent an important risk factor for developing reactions to these drugs. Paracetamol and nimesulide are relatively safe alternative choices in those patients, although their use still carries some risk of unwanted reactions. PMID- 10674574 TI - Adherence to antiretroviral therapy by pregnant women infected with human immunodeficiency virus: a pharmacy claims-based analysis. AB - OBJECTIVE: To assess adherence to antiretroviral therapy with the use of Medicaid pharmacy claims data for human immunodeficiency virus (HIV)-infected pregnant women and to identify associated maternal and health care factors. METHODS: We retrospectively studied a cohort of 2714 HIV-infected women in New York State who delivered live infants from 1993-96. Among 682 women prescribed antiretroviral therapy in the last two trimesters, we studied 549 who started therapy more than 2 months before delivery. Adherence was defined as adequate if the supplied drug covered at least 80% of the days from the first prescription in the last two trimesters until delivery. Multivariable analyses were used to examine associations between maternal and health care factors and adherence. RESULTS: Only 34.2% of 549 subjects had at least 80% adherence based on pharmacy data, a rate that remained stable over time. The adjusted odds ratios (ORs) of adherence for black (OR 0.47, 95% confidence interval [CI] 0.30, 0.75) and Hispanic (OR 0.49, 95% CI 0.29, 0.82) women were nearly 50% lower than for white women. The OR of adherence was 0.32 (95% CI 0.12, 0.90) for teenagers compared with women aged 25-29 years and 0.56 (95% CI 0.34, 0.92) for women in New York City versus those residing elsewhere. Women on antiretroviral therapy before pregnancy were more likely to adhere (OR 1.55, 95% CI 1.02, 2.35). CONCLUSION: Teenagers, women of minority groups, and women living in New York City had greater risks of poor antiretroviral adherence, whereas women already prescribed antiretrovirals before pregnancy had better adherence. Our conservative pharmacy data-based measure showed that most HIV-infected women adhered poorly and adherence did not improve over the 4-year study. PMID- 10674575 TI - Respiratory distress syndrome and maternal birth weight effects. AB - OBJECTIVE: To study traditional risk factors and the intergenerational risk factor maternal low birth weight (LBW) for respiratory distress syndrome (RDS) in infants in multiple ethnic groups. METHODS: The population-based database consists of hospital records linked to Washington state maternal and infant vital records. Four racial-ethnic groups were studied, whites, blacks, Native Americans, and Hispanics. Poisson regression models were used to estimate relative risks of various factors for RDS. RESULTS: Rates for RDS were whites 1.2%, blacks 1.9%, Native Americans 1.3%, and Hispanics 1.0%. Maternal LBW was associated with increased relative risk (RR) for RDS in whites (2.6, 95% confidence interval [CI] 1.6, 4.2) and blacks (3.3, 95% CI 1.9, 5.6) for infants born vaginally. Compared with mothers of normal infants, birth weights of mothers of infants with RDS and delivered vaginally were significantly lower in whites, blacks, and Native Americans. The association of maternal LBW with RDS persisted in blacks even when multiple risk factors were added to the model (RR 2.4; 95% CI 1.1, 5.1). CONCLUSION: The association of maternal LBW with RDS is probably due in part to the association of maternal LBW with infant LBW and preterm birth. The strong persistent association of maternal LBW with RDS in blacks suggests that improvement of perinatal outcomes in that group will require improvement of long term birth weight distribution. PMID- 10674576 TI - Plasma thiol status in preeclampsia. AB - OBJECTIVE: To measure plasma thiol levels in women with normal pregnancies, women with preeclampsia, and nonpregnant controls to define plasma thiol's effect on glutathione homeostasis and pathophysiology of preeclampsia. METHODS: Total plasma cysteine, gamma-glutamylcysteine, homocysteine, cysteinylglycine, and glutathione levels were measured in ten nonpregnant women, ten women with normotensive pregnancies, and 20 women with preeclampsia at delivery. RESULTS: Median total plasma levels of all thiols in normotensive pregnant women were significantly lower than in nonpregnant women. Median total plasma cysteine and homocysteine levels in women with preeclampsia were significantly higher compared with pregnant controls (254 versus 190 micromol/L, P < .001; and 13.3 versus 8.4 micromol/L, P < .02, respectively), whereas glutathione levels were significantly lower in women with preeclampsia compared with those in pregnant controls (5.1 versus 6.3 micromol/L, P < .05). CONCLUSION: In women with preeclampsia, homocysteine and cysteine levels, which are lowered in normotensive pregnancy, were comparable to levels in nonpregnant women, whereas glutathione levels were lower. Those results suggest that in women with preeclampsia, glutathione use is higher or its synthesis is disturbed. Therefore, glutathione might affect pathophysiology of preeclampsia. PMID- 10674577 TI - Serum insulin-like growth factor binding protein-1 at 16 weeks and subsequent preeclampsia. AB - OBJECTIVE: To determine whether serum concentrations of insulin-like growth factor-binding protein-1 (IGFBP-1), a major decidual protein, at 16 weeks' gestation differ between women who later develop pregnancy-related hypertension and normotensive women. METHODS: Concentrations of IGFBP-1 were measured using immunoenzymometric assay in serum samples collected for alpha-fetoprotein (AFP) and free beta subunit of hCG (free beta-hCG) determinations in a Down syndrome screening program at 16 weeks' gestation in a population-based cohort of 1049 nulliparous women. After exclusion of subjects with multiple pregnancies, insulin dependent diabetes, major fetal malformations, and incomplete data, 917 subjects remained eligible. RESULTS: The mean levels (+/- standard deviation) of IGFBP-1 were significantly lower in 34 women who later developed preeclampsia (73 +/- 43 microg/L, P < .01) and in 80 women with White A diabetes (84.7 +/- 53 microg/L, P < .01) compared with controls (103 +/- 58 microg/L). In seven women with White A diabetes and subsequent preeclampsia IGFBP-1 levels were especially low (41 +/- 34 microg/L). The concentrations of AFP and free beta-hCG in the subgroups with hypertensive disorders were not significantly different from those of normotensive women. CONCLUSION: Decreased IGFBP-1 levels at 16 weeks' gestation in women who develop preeclampsia might indicate impaired decidual function. Hyperinsulinemia, a known risk factor for preeclampsia, might contribute to decreased concentrations of serum IGFBP-1. However, due to low sensitivity, assay of serum IGFBP-1 was not clinically valuable for predicting preeclampsia. PMID- 10674578 TI - Glycemic control throughout pregnancy and fetal growth in insulin-dependent diabetes. AB - OBJECTIVE: To determine the time of growth acceleration in fetuses of insulin dependent diabetic women who are large for gestational age (LGA) at birth and the relationship between growth acceleration and diabetic control throughout pregnancy. METHODS: We studied a consecutive sample of 76 women with insulin dependent diabetes divided by those who delivered LGA or normally grown infants. Fetal abdominal circumference (AC) was measured ultrasonically at regular intervals between 20 and 34 weeks' gestation. Diabetic control was assessed by regular measurement of glycosylated hemoglobin and capillary blood glucose levels. RESULTS: A significant difference in fetal AC between groups developed between 20 and 24 weeks' gestation, and the LGA group continued to have accelerated fetal growth. Between 18 and 24 weeks glycosylated hemoglobin and capillary blood glucose concentrations were significantly higher in women who delivered LGA infants. After 28 weeks, blood glucose concentrations and glycosylated hemoglobin did not differ significantly between groups. There was a nonsignificant trend toward more vaginal deliveries in the normal group (45% versus 32%). CONCLUSION: In insulin-dependent diabetic pregnancy, although actual growth acceleration occurred from about 20 weeks' gestation, growth potential of fetuses appeared to be determined by prevailing maternal glucose concentrations before then. Excessive growth continued despite subsequent satisfactory glucose control. If strict blood glucose control is maintained during first and second trimesters, it might reduce the incidence of LGA infants. PMID- 10674579 TI - Fetal pancreatic function in infants of diabetic and rhesus-isoimmunized women. AB - OBJECTIVE: To measure insulin and glucagon concentrations in amniotic fluid (AF) collected near term in basal conditions and after an arginine test in diabetic, rhesus-isoimmunized, and control pregnant women. METHODS: At baseline, AF was collected from 44 diabetic, 32 rhesus-isoimmunized, and 27 control pregnant women in late pregnancy. Fifty-two diabetic, six rhesus-isoimmunized, and nine control pregnant women had amniocentesis 2 hours after arginine infusion (30 g intravenous/30 minutes) at 33-36 weeks. RESULTS: Baseline AF glucose concentrations were significantly greater in diabetic women than the other conditions, and they related to the gestational age in the women with hemolytic disease of the newborn. Insulin and glucagon AF content of isoimmunized pregnancies overlapped controls, whereas insulin and insulin/glucagon molar ratios were significantly higher, and glucagon values lower, in diabetic pregnancies compared with isoimmunized and control pregnancies. In isoimmunized pregnancies, the AF concentrations of glucose, insulin, and glucagon were correlated with gestational age (less than 34, 34 weeks or more). The samples collected after arginine infusion, compared with those collected at baseline, showed significantly greater insulin and insulin/glucagon molar ratio values in diabetic (28 +/- 5 versus 11 +/- 1 microU/mL, P = .001; 29.4 +/- 1.7 versus 12.0 +/- 2.8, P = .001) and in Rh pregnant women (18 +/- 6 versus 7.7 +/- 0.7 microU/mL, P = .001; 30 +/- 9 versus 3.4 +/- 0.4 I/G, P = .001), whereas no significant difference was observed in the controls. CONCLUSION: Basal islet hormone concentrations in AF are modified by maternal diabetes and further influenced by arginine administration. Arginine produces an AF response that is similar in pregnancies complicated by diabetes mellitus and rhesus isoimmunization, despite different (hyperglycemia and euglycemia) maternal blood glucose levels. PMID- 10674580 TI - The appropriateness of recommendations for hysterectomy. AB - OBJECTIVE: To evaluate the appropriateness of recommendations for hysterectomies done for nonemergency and non-oncologic indications. METHODS: We assessed the appropriateness of recommendations for hysterectomy for 497 women who had the operation between August 1993 and July 1995 in one of nine capitated medical groups in Southern California. Appropriateness was assessed using two sets of criteria, the first developed by a multispecialty expert physician panel using the RAND/University of California-Los Angeles appropriateness method, and the second consisting of the ACOG criteria sets for hysterectomies. The main outcome measure was the appropriateness of recommendation for hysterectomy, based on expert panel ratings and ACOG criteria sets. RESULTS: The most common indications for hysterectomy were leiomyomata (60% of hysterectomies), pelvic relaxation (11%), pain (9%), and bleeding (8%). Three hundred sixty-seven (70%) of the hysterectomies did not meet the level of care recommended by the expert panel and were judged to be recommended inappropriately. ACOG criteria sets were applicable to 71 women, and 54 (76%) did not meet ACOG criteria for hysterectomy. The most common reasons recommendations for hysterectomies considered inappropriate were lack of adequate diagnostic evaluation and failure to try alternative treatments before hysterectomy. CONCLUSION: Hysterectomy is often recommended for indications judged inappropriate. Patients and physicians should work together to ensure that proper diagnostic evaluation has been done and appropriate treatments considered before hysterectomy is recommended. PMID- 10674581 TI - Cost-effectiveness of deep venous thrombosis prophylaxis in gynecologic oncology surgery. AB - OBJECTIVE: To estimate the cost-effectiveness of preventive strategies for deep vein thrombosis (DVT) in patients undergoing surgery for gynecologic cancer. METHODS: A model was constructed to estimate the costs and outcomes associated with the use of external pneumatic compression, unfractionated heparin, and low molecular weight heparin in women with cervical, endometrial, and ovarian cancer. We estimated cost per DVT prevented, per fatal pulmonary embolus (PE) prevented, and per life-year saved. Probability estimates for various outcomes and efficacies were obtained from the literature, using data specific for gynecologic patients when available. RESULTS: Cost-effectiveness estimates ranged from $27 per life-year saved for a 55-year-old endometrial cancer patient to $5132 per life-year saved for a 65-year-old with ovarian cancer. Although low molecular weight heparin and unfractionated heparin were cost-effective compared with no prophylaxis, each was less effective than external pneumatic compression in the base case. The results of the analysis were sensitive to assumptions about the relative risk of DVT, the life expectancy of the patient, the costs of future treatment, and the relative effectiveness of the different strategies: If unfractionated heparin or low molecular weight heparin is at least 2-3% more effective than external pneumatic compression, then the incremental cost per life year of either would be less than $50,000 compared with external pneumatic compression. CONCLUSION: Prophylaxis of DVT is cost-effective in terms of life years gained even for patients with relatively short life expectancies, such as ovarian cancer patients. External pneumatic compression appears to be the most cost-effective strategy under our baseline assumptions, but further studies in gynecologic cancer are needed to validate our conclusions. PMID- 10674582 TI - Determinants of unexplained antepartum fetal deaths. AB - OBJECTIVE: To assess fetal, maternal, and pregnancy-related determinants of unexplained antepartum fetal death. METHODS: We conducted a hospital-based cohort study of 84,294 births weighing 500 g or more from 1961-1974 and 1978-1996. Unexplained fetal deaths were defined as fetal deaths occurring before labor without evidence of significant fetal, maternal, or placental pathology. RESULTS: One hundred ninety-six unexplained antepartum fetal deaths accounted for 27.2% of 721 total fetal deaths. Two thirds of the unexplained fetal deaths occurred after 35 weeks' gestation. The following factors were independently associated with unexplained fetal death: maternal prepregnancy weight greater than 68 kg (adjusted odds ratio [OR] 2.9; 95% confidence interval [CI] 1.85, 4.68), birth weight ratio (defined as ratio of birth weight to mean weight for gestational age) between 0.75 and 0.85 (OR 2.77; 95% CI 1.48, 5.18) or over 1.15 (OR 2.36; 95% CI 1.26, 4.44), fewer than four antenatal visits in women whose fetuses died at 37 weeks or later (OR 2.21; 95% CI 1.08, 4.52), primiparity (OR 1.74; 95% CI 1.26, 2.40), parity of three or more (OR 2.01; 95% CI 1.26, 3.20), low socioeconomic status (OR 1.59; 95% CI 1.14, 2.22), cord loops (OR 1.75; 95% CI 1.04, 2.97) and, for the 1978-1996 period only, maternal age 40 years or more (OR 3.69; 95% CI 1.28, 10.58). Trimester of first antenatal visit, low maternal weight, postdate pregnancy, fetal-to-placental weight ratio, fetal sex, previous fetal death, previous abortion, cigarette smoking, and alcohol use were not significantly associated with unexplained fetal death. CONCLUSION: In this study, we identified several factors associated with an increased risk of unexplained fetal death. PMID- 10674583 TI - Second-trimester cervical ultrasound: associations with increased risk for recurrent early spontaneous delivery. AB - OBJECTIVE: To determine whether short cervical length or internal os funneling before 20 weeks' gestation predicts early preterm birth or pregnancy loss in women with at least one prior spontaneous early preterm birth. METHODS: Transvaginal cervical ultrasound examinations were done every 2 weeks on 69 women with singleton gestations and histories of at least one prior spontaneous birth between 16 and 30 weeks' gestation. The results of those examinations were correlated with gestational age at delivery. RESULTS: Among 53 women who had ultrasound examinations before 20 weeks' gestation, those with cervical lengths at or below the tenth percentile for the study population (22 mm, n = 4) or funneling of the internal os (n = 5) were more likely than women without those factors to have spontaneous preterm births within 2 weeks (33% versus 0%, P = .01) or 4 weeks from the ultrasound examination (67% versus 0%, P < .001) or before 35 weeks' gestation (100% versus 19%, P < .001). Short cervical length or funneling between 20-24 and 25-29 weeks was also associated with increased risk of spontaneous preterm birth before 35 weeks' gestation (P < or = .05 and P = .002, respectively) but not with increased risk of spontaneous preterm birth within 2 or 4 weeks of ultrasound examination. CONCLUSION: Women with prior early spontaneous preterm births who have short cervical lengths or funneling of the internal cervical os before 20 weeks' gestation are at increased risk of subsequent spontaneous preterm birth. PMID- 10674584 TI - Progesterone, inhibin, and hCG multiple marker strategy to differentiate viable from nonviable pregnancies. AB - OBJECTIVE: To determine whether a combination of serum and urine biomarkers drawn from symptomatic pregnant women will help early differentiation of viable from nonviable pregnancies. METHODS: We conducted a prospective cohort study of 220 women who presented in the first trimester of pregnancy with complaints of pain, cramping, bleeding, or spotting. Serum samples for progesterone, inhibin A, and hCG, and urine beta-core hCG, were collected at presentation. To evaluate whether those biomarkers could predict viable and nonviable outcomes in pregnancy, we used likelihood ratios to compare operating characteristics of single and multiple biomarker strategies. RESULTS: Of 220 pregnancies studied, 98 were viable and 122 nonviable. Among single biomarkers, progesterone alone appears to have the greatest utility (area under the receiver operator characteristic curve = 0.923). Among dual-biomarker strategies, progesterone plus hCG and progesterone plus inhibin A improved specificity but not sensitivity. At 95% sensitivity, the combination of progesterone and hCG improved specificity from 0.29 to 0.66 (improvement = 0.37 [95% confidence interval 0.23, 0.52]). A triple-biomarker combination did not show substantial improvement over the dual-biomarker strategy. Also, combinations that used urine beta-core hCG did not improve diagnostic accuracy. CONCLUSION: Serum progesterone appeared to be the single most specific biomarker for distinguishing viable from nonviable pregnancies. When a dual-biomarker strategy was applied, combining serum progesterone with hCG, specificity improved significantly, which suggests that a multiple biomarker strategy might help distinguish viable from nonviable pregnancies in early gestation. PMID- 10674585 TI - Misoprostol's effect on uterine arterial blood flow and fetal heart rate in early pregnancy. AB - OBJECTIVE: To determine whether a single oral dose of misoprostol is associated with change in Doppler resistance indices (RIs) of the uterine artery in early pregnancy. METHODS: Forty pregnant women seeking legal termination of pregnancy at 7-15 completed gestational weeks were each given a single oral dose of 200 microg misoprostol. Resistance indices (A/B ratio) and pulsatility index (PI) of the uterine arteries (UA) and fetal heart rate (FHR) were assessed by Doppler ultrasound before and 1 hour after administration of misoprostol. RESULTS: Doppler RIs (UA-A/B and UA-PI) of the right and left uterine arteries increased significantly 1 hour after misoprostol administration. The right UA-A/B increased from 7.16 +/- 1.09 (mean +/- SEM) to 10.26 +/- 0.67 (P < .001), and the left UA A/B increased from 7.40 +/- 0.72 to 9.21 +/- 0.82 (P = .04). The right UA-PI increased from 2.38 +/- 0.11 to 2.90 +/- 0.12 (P < .001), and the left UA-PI increased from 2.38 +/- 0.17 to 2.70 +/- 0.18 (P = .03). No significant changes in FHR were noted 1 hour after misoprostol administration. None of the fetuses died during that time. CONCLUSION: Doppler RIs of the uterine arteries increased significantly after single oral doses of misoprostol during the first trimester, implying a reduction in arterial blood flow. Those changes were not associated with fetal death, possibly explaining congenital abnormalities associated with misoprostol in early pregnancy. PMID- 10674586 TI - Coccidioidomycosis in pregnancy during an epidemic in California. AB - OBJECTIVE: To determine presentation, clinical course, and outcome of a cohort of pregnant women with coccidioidomycosis and compare findings with common observations reported in the literature. METHODS: Thirty-two women who delivered live infants or aborted fetuses in 1993 and had confirmed diagnoses of coccidioidomycosis were included in the study. Medical records were evaluated retrospectively for clinical characteristics, laboratory results, and disease course. RESULTS: Dissemination occurred in three of 32 cases. The most common management was supportive and symptomatic care. At 1 year, 26 of 32 had recovered. There were no maternal deaths. CONCLUSION: The common depiction of coccidioidomycosis in pregnancy has overstated morbidity and mortality likely because of reporting bias. Many women will have favorable outcomes without drug treatment, and the practice of abortions or early delivery in subjects with active infection should be rare. PMID- 10674587 TI - Amniochorion gelatinase-gelatinase inhibitor imbalance in vitro: a possible infectious pathway to rupture. AB - OBJECTIVE: To estimate the effect of lipopolysaccharide on gelatinases and tissue inhibitors of matrix metalloproteinase 2 (gelatinase inhibitor) balance in human fetal membranes. METHODS: Amniochorionic membranes in organ explant were stimulated with 1000 ng/mL lipopolysaccharide for 24 hours after a 48-hour preincubation period. Quantitative competitive polymerase chain reaction (PCR) was done to quantitate messenger RNAs for gelatinase A and B (matrix metalloproteinase 2 and 9) and tissue inhibitor of metalloproteinase 2. Protein levels were assayed by enzyme-linked immunosorbant assay. The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 was calculated. Statistical evaluation was done by Mann-Whitney U test. RESULTS: Lipopolysaccharide stimulation produced 3.6 x 10(6) and 366 transcripts of gelatinase A and B, respectively, compared with only 5.9 x 10(4) (P = .009) and three transcripts (P = .006), respectively, in the controls. Lipopolysaccharide stimulation released 210 ng/mL compared with 7 ng/mL of gelatinase A and B proteins compared with 120 (P = .01) and 4.6 ng/mL (P = .3) in controls, respectively. Control amniochorion produced 5.7 x 10(5) transcripts of tissue inhibitor of metalloproteinase 2, whereas lipopolysaccharide stimulation produced 4.1 x 10(5) transcripts (P = .69). Lipopolysaccharide reduced the release of this inhibitor from 114 ng/mL to 68 ng/mL (P = .007). The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 increased from a balanced ratio of 1:1 to 3.1:1 after 1000 ng/mL of lipopolysaccharide. CONCLUSION: Lipopolysaccharide increased the expression and release of gelatinases and decreased its inhibitor, which shifted the balance in favor of gelatinase activity leading to membrane degradation that predisposes to premature rupture of membranes. PMID- 10674588 TI - Diet and sex-hormone binding globulin, dysmenorrhea, and premenstrual symptoms. AB - OBJECTIVE: To test the hypothesis that a low-fat, vegetarian diet reduces dysmenorrhea and premenstrual symptoms by its effect on serum sex-hormone binding globulin concentration and estrogen activity. METHODS: In a crossover design, 33 women followed a low-fat, vegetarian diet for two menstrual cycles. For two additional cycles, they followed their customary diet while taking a supplement placebo pill. Dietary intake, serum sex-hormone binding globulin concentration, body weight, pain duration and intensity, and premenstrual symptoms were assessed during each study phase. RESULTS: Mean (+/- standard deviation [SD]) serum sex hormone binding globulin concentration was higher during the diet phase (46.7 +/- 23.6 nmol/L) than during the supplement phase (39.3 +/- 19.8 nmol/L, P < .001). Mean (+/- SD) body weight was lower during the diet (66.1 +/- 11.3 kg) compared with the supplement phase (67.9 +/- 12.1 kg, P < .001). Mean dysmenorrhea duration fell significantly from baseline (3.9 +/- 1.7 days) to diet phase (2.7 +/- 1.9 days) compared with change from baseline to supplement phase (3.6 +/- 1.7 days, P < .01). Pain intensity fell significantly during the diet phase, compared with baseline, for the worst, second-worst, and third-worst days, and mean durations of premenstrual concentration, behavioral change, and water retention symptoms were reduced significantly, compared with the supplement phase. CONCLUSION: A low-fat vegetarian diet was associated with increased serum sex hormone binding globulin concentration and reductions in body weight, dysmenorrhea duration and intensity, and premenstrual symptom duration. The symptom effects might be mediated by dietary influences on estrogen activity. PMID- 10674589 TI - Menstrual blood loss measured 5-6 years after endometrial ablation. AB - OBJECTIVE: To examine objectively the long-term efficacy of endometrial ablation for menorrhagia. METHODS: Thirty-nine women with menorrhagia due to ovulatory dysfunctional bleeding treated previously by rollerball ablation were followed up 5-6 years later. Menstrual blood in sanitary towels was measured with the alkaline hematin technique in 26 women who were still premenopausal and in whom menstrual blood loss had been measured before and immediately after the original ablation. RESULTS: Mean (+/- standard deviation [SD]) menstrual blood loss (per menstrual period) was reduced from 90 mL +/- 14.4 before ablation to 3.8 mL +/- 2.1 at 3 months, 1.8 mL +/- 1.0 at 6 months, and 3.3 mL +/- 1.3 at 5-6 years after ablation. In women who were still menstruating, the mean hemoglobin concentration rose significantly from 126 to 135 g/L (P = .022). CONCLUSION: Rollerball endometrial ablation is a highly effective long-term therapy for carefully selected women with menorrhagia due to ovulatory dysfunctional uterine bleeding. PMID- 10674590 TI - Cigarette smoking and epithelial ovarian cancer by histologic type. AB - OBJECTIVE: To examine cigarette smoking as a risk factor for different types of epithelial ovarian cancer. METHODS: We used data from the Cancer and Steroid Hormone Study, a multicenter, population-based, case control investigation. Cases were 447 women aged 20-54 years with diagnoses of epithelial ovarian cancer. Controls were 3868 women selected by random-digit dialing. Conditional logistic regression was used to obtain odds ratios (ORs) and 95% confidence intervals (CIs) as estimators of the relative risk of ovarian cancer. With age and study site as conditioning variables, OR point estimates were additionally adjusted for parity and use of oral contraceptives. RESULTS: The OR of mucinous epithelial ovarian cancer for women who had ever smoked was 2.3 (95% CI 1.4, 3.9) and for current smokers was 2.9 (95% CI 1.7, 4.9). The OR of mucinous tumors for current smokers was significantly elevated regardless of years since first cigarette or age at which women first smoked. The OR of mucinous tumors for current smokers increased slightly as cumulative pack-years of smoking increased, although the trend was not significant. Similar patterns of elevated risk were not observed among serous, endometrioid, or other histologic types. Odds ratio point estimates for former smokers were not significantly elevated for any histologic type. CONCLUSION: Current cigarette smoking was a risk factor for mucinous epithelial ovarian cancer, but not other histologic types. PMID- 10674591 TI - Hormone withdrawal symptoms in oral contraceptive users. AB - OBJECTIVE: To measure the timing, frequency, and severity of hormone-related symptoms in oral contraceptive (OC) users, specifically to compare active-pill with hormone-free intervals. METHODS: Using daily diaries, women recorded pelvic pain, bleeding, headaches, analgesic use, nausea or vomiting, bloating or swelling, and breast tenderness during active-pill intervals and hormone-free intervals. Participants either had no prior OC use, had taken OCs and were restarting, or had been taking OCs continuously for 12 months or longer. RESULTS: Two hundred sixty-two women, 26 with no previous OC use, 43 prior users, and 193 current users, provided daily records of hormone-related symptoms. Subjects with no prior OC use and prior users restarting were similar in no recent OC use, and because of the small sample, they were pooled for analysis as new-start OC users. Current users had patterns of symptoms that were more frequent during hormone free intervals than during the three active-pill weeks. These included pelvic pain (70% versus 21%, P < .001), headaches (70% versus 53%, P < .001), use of pain medication (69% versus 43%, P < .001), bloating or swelling (58% versus 19%, P < .001), and breast tenderness (38% versus 16%, P < .001). Similar patterns were seen in new-start OC users after the first cycle. Among new-start OC users, menstrual flow patterns, headache, bloating or swelling, and breast-tenderness symptoms decreased during the three cycles to approach those levels of current users. CONCLUSION: Almost all symptoms assessed were significantly worse during the 7-day hormone-free interval than during the 21 days of hormone-containing pills. PMID- 10674592 TI - Access to emergency contraception. AB - OBJECTIVE: To evaluate access to emergency contraception among women seeking help from clinicians who registered to be listed on the Emergency Contraception Hotline (1-888-NOT-2-LATE, ie, 1-888-668-2528) and the Emergency Contraception Website (not-2-late.com). METHODS: Two college-educated investigators posing as women who had a condom break the previous night called 200 providers to seek help. RESULTS: Only 76% of attempts resulted in an appointment or telephone prescription from a hotline provider within 72 hours, 14% were failures, and 11% resulted in referrals to other providers not listed on the hotline or website. CONCLUSION: Even under ideal conditions, access to emergency contraception is currently constrained. Although emergency contraception could reduce significantly the incidence of unintended pregnancy and the consequent need for abortion, its potential will not be realized unless women have better access to clinicians who can prescribe emergency contraceptive pills. PMID- 10674593 TI - Meclizine for prevention of nausea associated with use of emergency contraceptive pills: a randomized trial. AB - OBJECTIVE: We conducted a randomized trial to determine whether pretreatment with meclizine reduces the incidence of nausea and vomiting associated with the Yuzpe regimen of emergency contraception. METHODS: We randomly assigned 343 women aged 18-45 years who were not at risk for pregnancy to pretreatment with 50 mg of meclizine, placebo, or no drug 1 hour before the first of two doses of emergency contraceptive pills. We asked participants to complete three questionnaires over the following 48 hours. RESULTS: The incidence of nausea was 47% in the group pretreated with meclizine and 64% in the other two groups (relative risk adjusted for center 0.7, 95% confidence intervals 0.6, 0.9 for comparisons of meclizine with both placebo and no drug). The severity of nausea and the incidence of vomiting were also significantly lower in the meclizine pretreatment group than in the other two groups. Drowsiness was reported by about twice as many women in the meclizine pretreatment group (31%) than in the other two groups (13% in the placebo group, 16% in the no-pretreatment group; P < .01 for both comparisons). CONCLUSION: Meclizine is effective for preventing nausea and vomiting associated with the Yuzpe regimen of emergency contraceptive pills. Women using this drug should be cautioned to anticipate drowsiness. PMID- 10674594 TI - Estrogen effects on postural balance in postmenopausal women without vasomotor symptoms: a randomized masked trial. AB - OBJECTIVE: To assess whether estrogen treatment given to postmenopausal women without vasomotor symptoms improves balance more than placebo. METHODS: Forty healthy postmenopausal women without vasomotor symptoms were randomized to transdermal 17beta-estradiol (E2) 50 microg/day for 14 weeks or identical transdermal placebo patches. Postural balance was measured with dynamic posturography before and after 4, 12, and 14 weeks of therapy. In this test, the visual, vestibular, and somatosensory systems were provoked with increasing difficulty and body sway was measured with a dual forceplate. A low score showed large sway and a score of 100 showed no sway at all. RESULTS: Thirty-eight women completed the study. Both groups had normal balance for their ages and near maximum scores in the three easier balance tests at baseline. In the most difficult test, both groups improved their postural balance significantly (from 13 to 32 and from 22 to 39, respectively) after 4 weeks. Thereafter, no change was seen. One problem was low statistical power, but the relative change in balance did not differ between groups. The comparison did not show even a minute advantage of E2 over placebo, so a study with higher power would probably not have shown a more pronounced effect of estrogen than placebo. The change over time did not differ between groups, which indicates a significant learning effect. CONCLUSION: In women without vasomotor symptoms, estrogen therapy did not seem to increase postural balance significantly more than placebo. However, we could not rule out that estrogens affect postural balance in women with vasomotor symptoms. PMID- 10674596 TI - The effect of transport on the rate of severe intraventricular hemorrhage in very low birth weight infants. AB - OBJECTIVE: To determine the incidence of grade III or IV intraventricular hemorrhage in very low birth weight (VLBW) infants born at level I hospitals and transported to one tertiary center compared with those delivered at the same level III facility. METHODS: We evaluated all newborns admitted to a large tertiary neonatal intensive care unit from June 1, 1992, through December 31, 1995. All live born infants with birth weights of 500-1200 g and at least 24 weeks' gestation were included. Neonatal transports within 24 hours of delivery from 11 level I facilities were compared with those delivered at the same level III center with respect to grade III and IV intraventricular hemorrhage. Various antenatal and neonatal data were collected. RESULTS: Thirty-seven newborns (11%) experienced grade III or IV intraventricular hemorrhages among 329 who met study criteria. There were 27 cases (9%) in the 285 inborn neonates compared with 10 of 44 outborn cases (23%) (P < .02, 95% confidence interval 0.15, 0.87). The mean gestational age of the neonates with grade III or IV intraventricular hemorrhages was significantly lower in the inborn group, which further emphasizes the finding. No other study factors explained the difference. CONCLUSION: We found a higher risk for grade III or IV intraventricular hemorrhage developing in VLBW infants born at level I hospitals and transported to the tertiary care center compared with those born at the level III facility. This data should be considered when analyzing the potential effects of perinatal deregionalization. PMID- 10674595 TI - 17 Beta-estradiol-stimulated nitric oxide production by neutrophils: effect on platelet activation. AB - OBJECTIVE: To evaluate the effect of 17beta-estradiol (E2) on the ability of human neutrophils to produce nitric oxide (NO) and its effects on platelet activation. METHODS: The expression of neuronal nitric oxide synthase (nNOS) protein and the formation of NO by 17beta-E2-incubated neutrophils from men were studied in vitro (ten male volunteers, no medical-surgical antecedents, aged 25 45 years). Platelet aggregometry and changes in cyclic guanosine monophospate (cGMP) levels were used to bioassay the functionality of NO released from neutrophils. RESULTS: Incubation of neutrophils derived from men with physiologic concentrations of 17beta-E2 (10(-10) to 10(-8) mol/L) enhanced the expression of nNOS protein. 17Beta-E2-incubated neutrophils also showed a significant increase in their ability to generate NO measured by the conversion of [3H]-L-arginine to [3H]-L-citrulline. Furthermore, 17beta-E2-incubated neutrophils showed a greater ability to prevent adenosine diphosphate (ADP)-induced platelet activation. Moreover, increased levels of cGMP were found in the coincubation of platelets with 17beta-E2-treated neutrophils. CONCLUSION: These results suggest that 17beta E2 increases the ability of human neutrophils to produce NO and therefore may contribute to cardiovascular disease protection. PMID- 10674597 TI - Office prenatal formula advertising and its effect on breast-feeding patterns. AB - OBJECTIVE: To compare the effect of formula company-produced materials about infant feeding to breast-feeding promotion materials without formula advertising on breast-feeding initiation and duration. METHODS: Five hundred forty-seven pregnant women were randomized to receive either formula company (commercial; n = 277) or specially designed (research; n = 270) educational packs about infant feeding at their first prenatal visit. Feeding method was determined at delivery. Breast-feeding duration of the 294 women who chose to breast-feed was ascertained at 2, 6, 12, and 24 weeks. Survival analyses were used to evaluate continuous outcomes, and chi2 and logistic regression analyses were used to evaluate discrete outcomes. RESULTS: Breast-feeding initiation (relative risk [RR] 0.93, 95% confidence interval [CI] 0.61, 1.43) and duration after 2 weeks (hazard ratio 1.19, 95% CI 0.86, 1.64) were not affected. Women in the commercial group were more likely to cease breast-feeding before hospital discharge (RR 5.80, 95% CI 1.25, 54.01) and before 2 weeks (adjusted odds ratio [OR] 1.91, 95% CI 1.02, 3.55). In subgroup analyses, women with uncertain goals for breast-feeding or goals of 12 weeks or less experienced shortened exclusive (hazard ratio 1.53, 95% CI 1.06, 2.21), full (hazard ratio 1.70, 95% CI 1.18, 2.48), and overall (hazard ratio 1.75, 95% CI 1.16, 2.64) breast-feeding duration when exposed to the commercial intervention. CONCLUSION: Although breast-feeding initiation and long term duration were not affected, exposure to formula promotion materials increased significantly breast-feeding cessation in the first 2 weeks. Additionally, among women with uncertain goals or breast-feeding goals of 12 weeks or less, exclusive, full, and overall breast-feeding duration were shortened. Educational materials about infant feeding should support unequivocally breast-feeding as optimal nutrition for infants; formula promotion products should be eliminated from prenatal settings. PMID- 10674598 TI - Endobag extractor to remove masses during laparoscopy. AB - BACKGROUND: Intra-abdominal masses are removed during laparoscopy using different types of endobags. However, in many cases the specimens are larger than the trocar or the incision in the abdomen, with a potential risk of endobag rupture. INSTRUMENT: We developed an instrument to facilitate extraction of an endobag during laparoscopy without the need for a conventional minilaparotomy. The endobag extractor has three removable diverging blades that symmetrically enlarge the operative canal in the abdominal wall if spread after sharp extension of the skin incision. The full endobag can be drawn through the canal without the risk of endobag rupture because the size of the canal can be individualized, building a funnel. EXPERIENCE: We removed various kinds of ovarian tumors, specimens from salpingo-oophorectomies, and other specimens in 22 cases. CONCLUSION: This new instrument allows easy removal of surgical specimens during laparoscopy without conventional minilaparotomy, regardless of the type of endobag used. We believe this instrument lessens the risk of endobag rupture. PMID- 10674599 TI - Computed tomography-guided pudendal block for treatment of pelvic pain due to pudendal neuropathy. AB - BACKGROUND: Severe pelvic pain secondary to pudendal neuropathy can be treated with repeated local anesthetic nerve blocks or with surgical decompression of the nerve. Computed tomographic (CT) needle guidance to identified reliable anatomic points might be useful for improved success rates. TECHNIQUE: A CT scan is used to determine baseline anatomy and identify the sacrospinous process. A metallic marker is used to create a perpendicular pathway from the sacrospinous process upward to the skin surface, where a local anesthetic is injected. A 22-gauge, 5 inch needle is inserted downward in a perpendicular direction to the target. Deep penetration and direction are confirmed by serial CT scans. Medication is injected and the needle is removed. EXPERIENCE: Twenty-six women with diagnoses of pudendal neuropathy were treated with injection therapy once per month, for five total treatments each. About three-quarters experienced improvement. There were no complications in this series. Outcomes were gratifying considering the complex patient population, all having failed multiple therapeutic trials. CONCLUSION: We believe this technique warrants further evaluation and application in instances where noninvasive therapy of pudendal neuropathy is indicated. PMID- 10674600 TI - An interactive web site for research on fetal heart rate monitoring. AB - BACKGROUND: Traditional methods of exchange of research information may not be rapid enough, especially for international multicenter studies or when discussing controversial issues such as the value of fetal monitoring. The Internet is a useful tool that provides numerous opportunities for immediate communication within a large and diverse community of researchers. METHOD: A Web site at http://www.sisporto.med.up.pt was developed for a multicenter research project with interlinked pages on automated fetal heart rate (FHR) monitoring. It includes background knowledge about the subject, detailed information about the project, and a few interactive pages. These pages allow online discussions, simulations of data analysis, and download of data for local FHR analysis. EXPERIENCE: The Web site has been accessed from all over the world. In particular, participating research centers have had easy and fast access to background project information, and a few other clinicians and researchers participated in our online discussions and used the simulation tools or the data provided for analysis of typical FHR patterns. CONCLUSION: Web sites can be useful in multicenter research projects and for scientific information exchange. PMID- 10674601 TI - Workforce projections for subspecialists in obstetrics and gynecology. AB - OBJECTIVE: To project the future supply of practicing subspecialists in obstetrics and gynecology based on the most recent numbers of physicians entering fellowships. METHODS: A discrete actuarial model was developed, and supply projections were examined using 1999 subspecialty fellowship numbers from the American Board of Obstetrics and Gynecology. RESULTS: The numbers of obstetrician gynecologists entering subspecialty fellowships in maternal-fetal medicine (MFM) and reproductive endocrinology-infertility (REI) declined sharply between 1994 and 1999. There was a slow increase in gynecologic oncology (GO) fellows. Projections show that the numbers of practicing MFM and GO subspecialists will double by 2020, but they will be serving a 20% larger female population in the United States. Numbers of practicing REI subspecialists will increase slowly. CONCLUSION: The number of fellows in GO continues to enlarge progressively though slightly, whereas those in MFM and REI have fallen sharply in recent years. Among four possible factors affecting growth or decline, the ones that seem most important are existing career opportunities for both generalist and subspecialist obstetrician-gynecologists and the length of subspecialty education. PMID- 10674602 TI - Professionalism in obstetrics-gynecology residency education: the view of program directors. AB - OBJECTIVE: To define the qualities of professionalism emphasized in obstetrics gynecology residencies and identify existing means of evaluating them. METHODS: A survey, designed to assess the importance of professionalism in residency programs and what means are utilized for its development, was sent to all 270 obstetrics-gynecology residency program directors in the United States. RESULTS: Two hundred thirteen surveys were returned (79%). Ninety-seven percent of all respondents indicated that they thought the development of professionalism was necessary for training obstetrics-gynecology residents, and 84.3% thought that formal educational training time should be devoted to this development. Over 85% endorsed faculty examples and mentoring as their methods of teaching professionalism. Respondents ranked honesty; accountability to patients, colleagues, and society; respect for patients; integrity; and excellence as the most important qualities of professionalism. Almost 79% believed those qualities were as important and as necessary as qualities of skill and knowledge in residency training. Almost 80% of respondents thought that the establishment of formal professionalism guidelines would be valuable in their training programs. CONCLUSION: A critical quality in resident education is professionalism, which receives emphasis in training programs largely through faculty example and mentoring. The variability inherent in such methods might be reduced by residency wide guidelines for uniform application of standards and to avoid arbitrariness in enforcement. PMID- 10674603 TI - Medical and osteopathic boards' positions on chaperones during gynecologic examinations. PMID- 10674604 TI - Eradication of Helicobacter pylori prevents recurrence of ulcer after simple closure of duodenal ulcer perforation: randomized controlled trial. AB - OBJECTIVE: In this randomized trial, the authors sought to determine whether eradication of Helicobacter pylori could reduce the risk of ulcer recurrence after simple closure of perforated duodenal ulcer. BACKGROUND DATA: Immediate acid-reduction surgery has been strongly advocated for perforated duodenal ulcers because of the high incidence of ulcer relapse after simple patch repair. Although H. pylori eradication is now the standard treatment of uncomplicated and bleeding peptic ulcers, its role in perforation remains controversial. Recently a high prevalence of H. pylori infection has been reported in patients with perforations of duodenal ulcer. It is unclear whether eradication of the bacterium confers prolonged ulcer remission after simple repair and hence obviates the need for an immediate definitive operation. METHODS: Of 129 patients with perforated duodenal ulcers, 104 (81%) were shown to be infected by H. pylori. Ninety-nine H. pylori-positive patients were randomized to receive either a course of quadruple anti-helicobacter therapy or a 4-week course of omeprazole alone. Follow-up endoscopy was performed 8 weeks, 16 weeks (if the ulcer did not heal at 8 weeks), and 1 year after hospital discharge for surveillance of ulcer healing and determination of H. pylori status. The endpoints were initial ulcer healing and ulcer relapse rate after 1 year. RESULTS: Fifty-one patients were assigned to the anti-Helicobacter therapy and 48 to omeprazole alone. Nine patients did not undergo the first follow-up endoscopy. Of the 90 patients who did undergo follow-up endoscopy, 43 of the 44 patients in the anti-Helicobacter group and 8 of the 46 in the omeprazole alone group had H. pylori eradicated; initial ulcer healing rates were similar in the two groups (82% vs. 87%). After 1 year, ulcer relapse was significantly less common in patients treated with anti Helicobacter therapy than in those who received omeprazole alone (4.8% vs. 38.1%). CONCLUSIONS: Eradication of H. pylori prevents ulcer recurrence in patients with H. pylori-associated perforated duodenal ulcers. Immediate acid reduction surgery in the presence of generalized peritonitis is unnecessary. PMID- 10674605 TI - Redefining the role of surgery for perforated duodenal ulcer in the Helicobacter pylori era. PMID- 10674606 TI - Gastroesophageal reflux disease in asthma: effects of medical and surgical antireflux therapy on asthma control. AB - OBJECTIVE: To critique the English-language reports describing the effects of medical and surgical antireflux therapy on respiratory symptoms and function in patients with asthma. METHODS: The Medline computerized database (1959-1999) was searched, and all publications relating to both asthma and gastroesophageal reflux disease were retrieved. RESULTS: Seven of nine trials of histamine receptor antagonists showed a treatment-related improvement in asthma symptoms, with half of the patients benefiting. Only one study identified a beneficial effect on objective measures of pulmonary function. Three of six trials of proton pump inhibitors documented improvement in asthma symptoms with treatment; benefit was seen in 25% of patients. Half of the studies reported improvement in pulmonary function, but the effect occurred in fewer than 15% of patients. In the one study that used optimal antisecretory therapy, asthma symptoms were improved in 67% of patients and pulmonary function was improved in 20%. Combined data from 5 pediatric and 14 adult studies of anti-reflux surgery indicated that almost 90% of children and 70% of adults had improvement in respiratory symptoms, with approximately one third experiencing improvements in objective measures of pulmonary function. CONCLUSIONS: Fundoplication has been consistently shown to ameliorate reflux-induced asthma; results are superior to the published results of antisecretory therapy. Optimal medical therapy may offer similar results, but large studies providing support for this assertion are lacking. PMID- 10674607 TI - Colon interposition for esophageal replacement: an alternative technique based on the use of the right colon. AB - OBJECTIVE: To describe the technique and results of an alternative colon interposition procedure in which the ascending and transverse colon is used as graft, but that still relies on the left colonic artery for blood supply. SUMMARY BACKGROUND DATA: The standard procedure to obtain a left colon interposition graft requires ligation of the middle colic artery and mobilization of the left and right flexure. This approach carries a risk because preparation of the left flexure may damage arterial or venous collaterals located at this site that are crucial for graft perfusion. METHODS: The authors modified the standard technique so that mobilization of the left flexure is no longer necessary. To obtain a colon interposition graft that is long enough, the ascending colon was included into the graft by ligating the middle and the right colic artery. The left colic artery remained the blood-supplying vessel. From January 1997 to June 1998, 15 patients underwent modified colon interposition with a cervical anastomosis (12 esophagectomies, 3 esophagogastrectomies). RESULTS: In all cases, intraoperative blood supply from the left colic artery to the proximal ascending colon was sufficient. After surgery, four major complications occurred (27%). Endoscopy demonstrated a vital graft in all patients. In one patient a leakage of the cervical anastomosis was observed. One patient died of herpes pneumonia. Postoperative artificial ventilation was required for an average of 2.8 +/- 4.6 days, the average intensive care unit stay was 6.9+/-4.5 days, and the average total hospital stay was 24.1 +/- 15.1 days. CONCLUSION: An intact left colic artery, including its collaterals at the splenic flexure, supplies sufficient blood to the proximal ascending colon after central ligation of the middle and right colic artery. Even without mobilization of the left flexure, a sufficient graft length can be obtained. Preliminary complication rates with the use of this technique for colon interposition are in the range of those found for the standard colon interposition technique. These modifications may represent an alternative to established procedures for creating a colon interposition graft. PMID- 10674608 TI - Clinical significance of p53 mutations in adenocarcinoma of the esophagus and cardia. AB - OBJECTIVE: To compare the frequency and spectrum of p53 gene mutations in adenocarcinomas of the esophagus and cardia and to compare clinical and pathologic features in patients with p53 mutant and nonmutant cancers. SUMMARY BACKGROUND DATA: The p53 gene is commonly mutated in human cancers, and a p53 mutation is reported to be present in more than 50% of esophageal adenocarcinomas. Although many studies have investigated the frequency of p53 protein overexpression in adenocarcinomas of the esophagus or esophagogastric junction, few studies have assessed the frequency and clinical significance of p53 mutations in these tumors. In particular, the prognostic importance of p53 mutation is uncertain. Adenocarcinomas of the esophagus and cardia share many epidemiologic and pathologic features, but it is controversial whether they represent the same tumor. A comparison of the frequency and spectrum of mutations in adenocarcinomas of the esophagus and cardia would test whether these tumors are also similar at the molecular level. METHODS: DNA was isolated from microdissected paraffin-embedded tumor tissues of patients who underwent esophagogastrectomy for adenocarcinoma of the esophagus (n = 19), cardia (esophagogastric junction, n = 12), or subcardia (n = 6). Exons 5 to 8 of the p53 gene were analyzed for the presence of mutations using the polymerase chain reaction with single-strand conformation polymorphism and DNA sequencing of bands showing abnormal mobility. The presence of mutation was confirmed by selective hybridization of a mutant-specific oligonucleotide to DNA isolated from the tumor. RESULTS: p53 mutations were identified in 18 of 37 (48.6%) tumors. Patients with p53 mutant tumors were significantly younger and had a significantly poorer prognosis. There was a similar prevalence of p53 mutations in adenocarcinomas of the esophagus (53%) and cardia (58%). In contrast, mutations were relatively uncommon in subcardia adenocarcinomas (one mutant tumor [17%]). The types of mutations found in the esophageal and the cardia cancers were also similar. CONCLUSIONS: Adenocarcinomas of the esophagus and cardia have a similar frequency and spectrum of p53 gene mutations, suggesting that these tumors have a common pathogenesis. Patients with mutations are younger, have signs of more advanced disease, and have a poorer prognosis than patients without mutations. PMID- 10674609 TI - Prognostic factors in gastric stump carcinoma. AB - OBJECTIVE: To compare prognostic results in patients with gastric stump cancer (GSC) versus those with primary gastric cancer (PGC). SUMMARY BACKGROUND DATA: Gastric stump carcinomas have often been described as having low resectability rates and a poor prognosis. METHODS: Results of surgical treatment of 50 patients with GSC were compared with that of 516 patients with PGC. RESULTS: The resectability rate was 94% for GSC patients and 96.5% for PGC patients, without significant differences in terms of postoperative complications, death rate, and median survival time (31.6 vs. 32.9 months). The multivariate analysis showed an independent prognostic effect for R0 resection, pT1 and pT2 category, and age older than 65 years. CONCLUSION: The prognosis after resection and adequate lymphadenectomy does not differ between patients with GSC and PGC. PMID- 10674610 TI - Effect of intravascular volume expansion on renal function during prolonged CO2 pneumoperitoneum. AB - OBJECTIVE: To evaluate whether intravascular volume expansion would improve renal blood flow and function during prolonged CO2 pneumoperitoneum. SUMMARY BACKGROUND DATA: Although laparoscopic living donor nephrectomies have a considerably reduced risk of complications for the donors, significant concerns exist regarding procurement of a kidney in the altered physiologic environment of CO2 pneumoperitoneum. Recent studies have documented adverse effects of CO2 pneumoperitoneum on renal hemodynamics. METHODS: Renal and systemic hemodynamics and renal histology were studied in a porcine CO2 pneumoperitoneum model. After placement of a pulmonary artery catheter, carotid arterial line, Foley catheter, and renal artery ultrasonic flow probe, CO2 pneumoperitoneum (15 mmHg) was maintained for 4 hours. Pigs were randomized into three intravascular fluid protocol groups: euvolemic (3 mLkg/hour isotonic crystalloid), hypervolemic (15 mL/kg/hour isotonic crystalloid), or hypertonic (3 mL/kg/hour isotonic crystalloid plus 1.2 mL/kg/hour 7.5% NaCl). RESULTS: In the euvolemic group, prolonged CO2 pneumoperitoneum caused decreased renal blood flow, oliguria, and impaired creatinine clearance. Both isotonic and hypertonic volume expansions reversed the changes in renal blood flow and urine output, but impaired creatinine clearance persisted. CONCLUSIONS: Intravascular volume expansion alleviates the effects of CO2 pneumoperitoneum on renal hemodynamics in a porcine model. Hypertonic saline (7.5% NaCl) solution may maximize renal blood flow in prolonged pneumoperitoneum, but it does not completely prevent renal dysfunction in this setting. This study suggests that routine intraoperative volume expansion is important during laparoscopic live donor nephrectomy. PMID- 10674611 TI - Stapled versus sutured closure of loop ileostomy: a randomized controlled trial. AB - OBJECTIVE: To compare the outcome after conventional sutured loop ileostomy closure with stapled ileostomy closure. SUMMARY BACKGROUND DATA: A defunctioning loop ileostomy is now widely used in colorectal surgery. Subsequent closure may be associated with early complications, particularly bowel obstruction. The results of a preliminary nonrandomized study suggested that there was no significant difference in the rate of complications between sutured and stapled closure of loop ileostomy. METHODS: One hundred forty-one consecutive patients who underwent loop ileostomy between 1993 and 1998 were randomized before surgery to either sutured or stapled loop ileostomy closure. Seventy-one patients had stapled closure and 70 had sutured closure. RESULTS: Both groups were comparable in terms of age, sex, original operation, duration after original operation, and level of operating surgeon. Postoperative bowel obstruction occurred in 10/70 (14%) patients after sutured closure compared with 2/71 (3%) patients after stapled closure. Subgroup analysis of ileostomy closure in patients having an ileal pouch showed no significant difference in bowel obstruction between stapled and sutured closure (2/30 vs. 7/29). The incidence of other complications, readmissions, and reoperations did not differ between the two groups. The stapled closure was only 4 minutes quicker than sutured closure. The mean total hospital stay tended to be shorter after the stapled closure than the sutured closure, but this did not reach statistical significance. CONCLUSIONS: Bowel obstruction occurred less frequently after stapled closure, but the mean hospital stay and readmission and reoperation rate did not significantly differ between the two groups. PMID- 10674612 TI - Clinical and pathologic correlation of 84 mucinous cystic neoplasms of the pancreas: can one reliably differentiate benign from malignant (or premalignant) neoplasms? AB - OBJECTIVE: To determine whether the long-term behavior of cystic mucinous neoplasms of the pancreas could be predicted using a novel, precisely defined classification of benign mucinous cystadenomas, noninvasive proliferative cystic mucinous neoplasms, and invasive mucinous cystadenocarcinomas. The primary interest was to obtain long-term follow-up after complete resection to determine the recurrence rates based on this objective classification. BACKGROUND: Current understanding is that all cystic mucinous neoplasms of the pancreas are potentially malignant and that mucinous cystadenomas, when completely removed, are biologically benign. Cystadenocarcinomas are thought to be less aggressively malignant than ordinary ductal adenocarcinoma, but reported recurrence rates vary widely and are unpredictable. METHODS: All patients who underwent "curative" resection for cystic mucinous neoplasms at Mayo Clinic Rochester from 1940 to 1997 were identified. All available pathology slides, gross specimens, and clinical records were reviewed, eliminating patients with inadequate documentation. Neoplasms were reclassified as mucinous cystadenomas, noninvasive proliferative mucinous cystic neoplasms, or invasive cystadenocarcinomas based on specific histologic criteria. RESULTS: Of 84 patients (70 women, 14 men) with cystic mucinous neoplasms of the pancreas, 54 were classified as cystadenomas, 23 as noninvasive proliferative cystic mucinous neoplasms, and only 7 as cystadenocarcinomas. Recurrent disease developed in none of the 77 patients without invasion, but 5 of the 6 patients surviving resection for cystadenocarcinomas died of recurrent cystadenocarcinoma within 5 years. CONCLUSIONS: When the neoplasm is completely resected and subjected to adequate histopathologic examination based on these objective criteria, absence of tissue invasion predicts a curative operation and detailed follow-up may be unnecessary. In contrast, a histologic diagnosis of invasive cystadenocarcinoma portends a dismal prognosis, similar to that of typical ductal adenocarcinoma of the pancreas. PMID- 10674614 TI - Relation among portal segmentation, proper hepatic vein, and external notch of the caudate lobe in the human liver. AB - OBJECTIVE: To identify portal segmentation and a portal fissure in the caudate lobe of the human liver in relation to the hepatic venous system and the external notch at the caudal edge of the caudate lobe. SUMMARY BACKGROUND DATA: Although the anatomy of the caudate lobe has been studied, the detailed anatomy has not yet been clarified; this is necessary to develop safe procedures for caudate lobe resection. METHODS: A total of 88 formalin-fixed human livers were dissected to visualize the portal vein and hepatic vein systems of the caudate lobe in relation to the external notch. RESULTS: The patterns of portal branching were classified into two types. In 58 livers (67.4%), the territories of the first order portal branches were clearly divided into two areas (the Spiegel lobe and the paracaval portion). In the remaining 28 livers (32.6%), the territories of the second-order portal branches were clearly divided into two areas. These two areas were distinctly separated by an internal plane, which was coincident with the external notch. The caudate lobe had a systematized hepatic venous system that consisted of one (87.5%) or two (11.4%) proper hepatic veins and plural accessory hepatic veins. The proper hepatic veins laid along the internal plane between these two portal areas. CONCLUSION: The caudate lobe exhibited distinct portal segmentation with a portal fissure that was indicated internally by the proper hepatic vein and externally by the notch at the caudal edge of the caudate lobe. PMID- 10674613 TI - Quantitative measurement of P- and E-selectin adhesion molecules in acute pancreatitis: correlation with distant organ injury. AB - OBJECTIVE: To determine whether expression of P- and E-selectin molecules is associated with the development of systemic organ manifestations in acute pancreatitis (AP). SUMMARY BACKGROUND DATA: Overproduction of inflammatory cytokines in AP induces expression of adhesion molecules, which may lead to increased leukocytic infiltration and tissue damage. Understanding the temporal expression of these molecules could afford better measures for therapeutic intervention. METHODS: Acute pancreatitis was induced in 30-day-old female C57/ bI/6J mice by feeding a choline-deficient/ethionine-supplemented diet (n = 95). Mice were divided into three groups. Group I (n = 35) was used to study the biochemical and histologic manifestations of AP and to evaluate the neutrophilic infiltration by myeloperoxidase activity and immunofluorescence. Groups II (n = 35) and III (n = 25) were used to evaluate expression of P- and E-selectin by the dual radiolabeled monoclonal antibody technique. RESULTS: Biochemical and histologic evidence of AP developed in all mice. The inflammatory cytokine tumor necrosis factor-alpha gradually increased in serum as early as 18 hours, reaching more than 800-fold background levels by 72 hours. Biphasic P-selectin expression in the lung was seen with peaks at 24 and 48 hours; E-selectin expression peaked at 48 hours. CD18-positive leukocytes and increased myeloperoxidase activity in the lung were demonstrated at 24 hours, correlating with the onset of selectin upregulation. Histologic scoring of lung tissue demonstrated mild damage at 24 hours, with progressive injury occurring from 48 to 72 hours. CONCLUSIONS: In AP, the production of inflammatory cytokines precedes up-regulation of P- and E selectin, whose expression coincided with the increased infiltration of CD18 positive cells and neutrophil sequestration in lung tissue. Temporally, these events correlate with evidence of histologic pulmonary injury and underscore the role of adhesion molecules as mediators of pathophysiologic events. This mechanistic pathway may afford novel therapeutic interventions in clinical disease by using blocking agents to ameliorate the systemic manifestations of AP. PMID- 10674615 TI - Preoperative evaluation of patients with primary head and neck cancer using dual head 18fluorodeoxyglucose positron emission tomography. AB - OBJECTIVE: To evaluate the value of 18fluorodeoxyglucose (FDG) positron emission tomography (PET) in primary head and neck cancer. BACKGROUND DATA: Head and neck carcinomas tend to metastasize to regional lymph nodes rather than to spread hematogenously. With nodal metastases, cure rates decrease by approximately 50%. Moreover, in approximately 3% of the patients, a second primary tumor is found at initial presentation. METHODS: Fifty-four consecutive patients (31 men and 23 women; mean age 60 years, range 34-81 years) with previously untreated squamous cell carcinomas of the oral cavity or oropharynx were studied. Before surgery and within a period of 3 weeks, clinical examination, chest x-ray, computed tomography (CT), ultrasonography with fine-needle aspiration cytology (US/ FNAC), and FDG-PET were performed. All study results were scored per neck side and were also classified as 0 (no metastases), 1 (single metastasis), or 2 (multiple metastases). RESULTS: The sensitivity for the detection of lymph node metastases per neck side was 96%, 85%, and 64% for FDG-PET, CT, and US/FNAC, respectively. The specificity was 90%, 86%, and 100% for FDG-PET, CT, and US/FNAC, respectively. In terms of the classification, FDG-PET showed the best correlation with the histologic data. Finally, in nine patients (17%), a second primary tumor was detected by FDG-PET and confirmed by histologic evaluation. CONCLUSION: Because of the high prevalence of second primary tumors detected by FDG-PET and the decreased error rate in the assessment of lymph node involvement compared with CT and US, FDG-PET should be routinely performed in patients with primary head and neck cancer. PMID- 10674616 TI - Mammographically detected ductal carcinoma in situ treated with conservative surgery with or without radiation therapy: patterns of failure and 10-year results. AB - OBJECTIVE: The authors reviewed their institution's experience treating mammographically detected ductal carcinoma in situ (DCIS) of the breast with breast-conserving therapy (BCT) to determine 10-year rates of local control and survival, patterns of failure, and factors associated with outcome. SUMMARY BACKGROUND DATA: From January 1980 to December 1993, 177 breasts in 172 patients were treated with BCT for mammographically detected DCIS of the breast at William Beaumont Hospital, Royal Oak, Michigan. METHODS: All patients underwent an excisional biopsy, and 65% were reexcised. Thirty-one breasts (18%) were treated with excision alone, whereas 146 breasts (82%) received postoperative radiation therapy (RT). All patients undergoing RT received whole-breast irradiation to a median dose of 50.0 Gy. One hundred thirty-six (93%) received a boost to the tumor bed for a median total dose of 60.4 Gy. Median follow-up was 5.9 years for the lumpectomy alone group and 7.2 years for the lumpectomy + RT group. RESULTS: In the entire population, 15 patients had an ipsilateral breast recurrence. The 5 and 10-year actuarial rates of ipsilateral breast recurrence were 7.8% and 7.8% for lumpectomy alone and 8.0% and 9.2% for lumpectomy + RT, respectively. Eleven of the 15 recurrences developed within or immediately adjacent to the lumpectomy cavity and were designated as true recurrences or marginal misses (TMM). Four recurred elsewhere in the breast. Eleven of the 15 recurrences were invasive, whereas 4 were pure DCIS. Only one patient died of disease, yielding 5- and 10 year actuarial cause-specific survival rates of 100% and 99.2%, respectively. Eleven patients were diagnosed with subsequent contralateral breast cancer, yielding 5- and 10-year actuarial rates of 5.1% and 8.3%, respectively. Clinical, pathologic, and treatment-related factors were analyzed for an association with ipsilateral breast failure or TR/MM. No factors were significantly associated with ipsilateral breast failure. In the entire population, the omission of RT and younger age at diagnosis were significantly associated with TR/MM. Patients younger than 45 years at diagnosis had a significantly higher rate of TR/MM in both the lumpectomy + RT and lumpectomy alone groups. None of the 37 patients who received a postexcisional mammogram had an ipsilateral breast failure versus 15 in the patients who did not receive a postexcisional mammogram. CONCLUSIONS: Patients diagnosed with mammographically detected DCIS of the breast appear to have excellent 100-year rates of local control and overall survival when treated with BCT. These results suggest that the use of RT reduces the risk of local recurrence and that patients diagnosed at a younger age have a higher rate of local recurrence with or without the use of postoperative RT. PMID- 10674617 TI - Insulinlike growth factor I plus insulinlike growth factor binding protein 3 attenuates the proinflammatory acute phase response in severely burned children. AB - OBJECTIVE: To determine the effect of insulinlike growth factor I (IGF-I) in combination with its principal binding protein (IGFBP-3) on the hepatic acute phase response in severely burned children. SUMMARY BACKGROUND DATA: The hepatic acute phase response is a cascade of events initiated to restore homeostasis after trauma. A prolonged response, however, may contribute to multiple organ failure, hypermetabolism, complications, and death. METHODS: Twenty-two children with a mean total body surface area (TBSA) burn of 57 +/- 3% were given a continuous infusion of 1 to 4 mg/kg/day IGF-I/BP-3 for 5 days after wound excision and grafting. Eight children with a TBSA burn of 54 +/- 4% were given saline as controls. Before and 5 days after excision and grafting, blood samples were taken for serum hepatic constitutive protein, acute phase protein, and proinflammatory cytokine analysis. RESULTS: Serum IGF-I levels in burned children given the IGF-I/BP-3 complex increased from 113 +/- 15 to 458 +/- 40 ng/mL and IGFBP-3 levels increased from 1.8 +/- 0.2 to 3.1 +/- 0.3 ng/mL. Levels of serum constitutive hepatic proteins (prealbumin, retinol-binding protein, and transferrin) increased with IGF-I/BP-3, whereas levels of type I acute phase proteins (C-reactive protein, alpha1-acid glycoprotein, and complement C-3) decreased when compared with controls. The complex had no effect on type II acute phase proteins. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL 1beta) levels decreased with IGF-I/BP-3 compared with controls, with no effect on interleukin-6. CONCLUSION: Severely burned children receiving IGF-I/BP-3 showed a decrease in IL-1beta and TNF-alpha followed by a decrease in type I acute phase proteins that was associated with a concomitant increase in constitutive hepatic proteins. Attenuating the proinflammatory acute phase with IGF-1/BP-3 response may prevent multiple organ failure and improve clinical outcomes after thermal injury without any detectable adverse side effects. PMID- 10674618 TI - Injury induces deficient interleukin-12 production, but interleukin-12 therapy after injury restores resistance to infection. AB - OBJECTIVE: To assess at serial intervals the production of interleukin-12 (IL-12) by monocytes/macrophages from the peripheral blood of injured patients and control subjects, and using a mouse model to confirm human findings and explore the effectiveness of low-dose IL-12 therapy in restoring resistance to infection after injury. SUMMARY BACKGROUND DATA: Serious injury is associated with loss of function of the T helper 1 lymphocyte phenotype, but little is known about IL-12 production in injured patients. The authors previously reported that early, moderate-dose IL-12 therapy in a mouse model of burn injury restored resistance to a later infectious challenge (cecal ligation and puncture, CLP). However, the efficacy of clinically relevant low-dose IL-12 therapy carried out to or beyond the time of septic challenge remains to be tested. METHODS: Peripheral blood mononuclear cells (PBMCs) and adherent cells were obtained from 27 patients with major burns or traumatic injury and 18 healthy persons and were studied at serial intervals for IL-12 production stimulated by bacterial lipopolysacharide (LPS). PBMCs from 18 of the same patients were studied for IL-10 production as well. IL 12 production by adherent cells from the spleens of burn or sham burn mice was studied at serial intervals after injury to confirm the human findings. Low-dose IL-12 or vehicle was given every other day to groups of burn and sham burn mice, which were then challenged with CLP on day 10, and survival was determined. Finally, spleens were harvested from burn or sham burn animals receiving low-dose IL-12 or vehicle after CLP. After splenic cellularity was determined by hemocytometer, splenocytes were cultured and production of tumor necrosis factor alpha, interferon-gamma, and IL-10 were assessed by immunoassay. RESULTS: Adherent cells from patients' PBMCs produced significantly less IL-12 than normal PBMCs after injury, reaching a nadir 8 to 14 days after injury. Stimulation of whole PBMCs by LPS indicated that at 8 to 14 days after injury, IL-12 production by PBMCs was significantly lower and IL-10 production was significantly higher than that of PBMCs from healthy persons. Low-dose IL-12 therapy significantly increased survival after CLP. Splenocytes from burn mice treated with IL-12 had significantly increased production of TNF-alpha and IF-beta, both before and after CLP, when compared with vehicle-treated burn animals. IL-10 production by bum splenocytes remained high after IL-12 treatment. Splenic cellularity increased after IL-12 treatment in burn mice. CONCLUSION: The capacity to produce IL-12 by adherent cells of the monocyte/macrophage lineage is significantly reduced after serious injury in humans and in a mouse burn model. In humans, there is a reciprocal relation between diminished IL-12 production and increased IL-10 production at approximately 1 week after injury. Low-dose IL-12 therapy in the mouse burn model markedly increased survival after a septic challenge, even when treatment was carried beyond the onset of sepsis. Low-dose IL-12 treatment in the mouse increased production of proinflammatory mediators important in host defense and at the same time maintained or increased production of IL-10, an important antiinflammatory cytokine. PMID- 10674619 TI - Survival and functional outcome after prolonged intensive care unit stay. AB - OBJECTIVE: To examine the functional outcome and costs of a prolonged illness requiring a stay in the surgical intensive care unit (SICU) of 7 of more days. SUMMARY BACKGROUND DATA: The long-term benefits and costs after a prolonged SICU stay have not been well studied. METHODS: All patients with an SICU length of stay of 7 or more days from July 1, 1996, to June 30, 1997, were enrolled. One hundred twenty-eight patients met the entry criteria, and mortality status was known in 127. Functional outcome was determined at baseline and at 1, 3, 6, and 12 months using the Sickness Impact Profile score, which ranges from 0 to 100, with a score of 30 being severely disabled. Hospital costs for the index admission and for all readmissions to Johns Hopkins Hospital were obtained. All data are reported as median values. RESULTS: For the index admission, age was 57 and APACHE II score was 23. The initial length of stay in the ICU was 11 days; the hospital length of stay was 31 days. The Sickness Impact Profile score was 20.2 at baseline, 42.9 at 1 month, 36.2 at 3 months, and 20.3 at 6 months, and was lower than baseline at 1 year. The actual 1-year survival rate was 45.3%. The index admission median cost was $85,806, with 65 total subsequent admissions to this facility. The cost for a single 1-year survivor was $282,618 (1996). CONCLUSIONS: An acute surgical illness that results in a prolonged SICU stay has a substantial in-hospital death rate and is costly, but the functional outcome from both a physical and physiologic standpoint is compatible with a good quality of life. PMID- 10674620 TI - Decreased surgical risks of pancreas transplantation in the modern era. AB - OBJECTIVE: To document the decreased incidence of surgical complications after pancreas transplantation in recent times. SUMMARY BACKGROUND DATA: Compared with other abdominal transplants, pancreas transplants have historically had the highest incidence of surgical complications. However, over the past few years, the authors have noted a significant decrease in the incidence of surgical complications. METHODS: The authors studied the incidence of early (<3 months after transplant) surgical complications (e.g., relaparotomy, thrombosis, infections, leaks) after 580 pancreas transplants performed during a 12-year period. Patients were analyzed and compared in two time groups: era 1 (June 1, 1985, to April 30, 1994, n = 367) and era 2 (May 1, 1994, to June 30, 1997, n = 213). RESULTS: Overall, surgical complications were significantly reduced in era 2 compared with era 1. The relaparotomy rate decreased from 32.4% in era 1 to 18.8% in era 2. Significant risk factors for early relaparotomy were donor age older than 40 years and recipient obesity. Recipients with relaparotomy had significantly lower graft survival rates than those without relaparotomy, but patient survival rates were not significantly different. A major factor contributing to the lower relaparotomy rate in era 2 was a significant decrease in the incidence of graft thrombosis; the authors believe this lower incidence is due to the routine use of postoperative low-dose intravenous heparin and acetylsalicylic acid. The incidence of bleeding requiring relaparotomy did not differ between the two eras. Older donor age was the most significant risk factor for graft thrombosis. The incidence of intraabdominal infections significantly decreased between the two eras; this decrease may be due to improved prophylaxis regimens in the first postoperative week. CONCLUSIONS: Although a retrospective study has its limits, the results of this study, the largest single-center experience to date, show a significant decrease in the surgical risk associated with pancreas transplants. Reasons for this decrease are identification of donor and recipient risk factors, better prophylaxis regimens, refinements in surgical technique, and improved immunosuppressive regimens. These improved results suggest that more widespread application of pancreas transplantation is warranted. PMID- 10674621 TI - Bactericidal activity against coagulase-negative staphylococci is impaired in infants receiving long-term parenteral nutrition. AB - OBJECTIVE: To examine the role of total parenteral nutrition (TPN) in predisposing infants to infection caused by coagulase-negative staphylococci. SUMMARY BACKGROUND DATA: Total parenteral nutrition is an important means of providing essential nutrients to newborn infants. However, its use has been associated with complications, particularly infection caused by coagulase negative staphylococci. Recent data suggest that TPN may modulate immune function; however, reports directly indicating impaired immunity against coagulase-negative staphylococci during TPN are limited. METHODS: Study 1 involved 31 infants younger than 4 months who had undergone surgery and were not receiving antibiotics; 20 were receiving TPN and 11 were receiving a normal enteral diet. An in vitro whole blood model was used to measure the host bactericidal activity against coagulase-negative staphylococci. Bacterial killing and phagocytosis were measured after a 45-minute challenge with viable coagulase negative staphylococci. In study 2, whole blood killing and intracellular killing of coagulase-negative staphylococci were measured in five newborn infants (younger than 2 months) who were receiving long-term TPN (>10 days), five control infants receiving a normal enteral diet, and five healthy adults. RESULTS: In study 1, infants receiving a normal enteral diet showed a high capacity to ingest and kill coagulase-negative staphylococci. In contrast, the blood of infants receiving long-term TPN showed a reduction in coagulase-negative staphylococci phagocytosis and killing. There were significant negative linear correlations between the duration of TPN and killing of coagulase-negative staphylococci and phagocytosis of coagulase-negative staphylococci. In study 2, infants receiving long-term TPN had lower whole blood killing and intracellular killing than infants receiving a normal enteral diet and healthy adult volunteers. These data seem to indicate a neutrophil dysfunction mediated by TPN in infancy. CONCLUSIONS: Host defense mechanisms, including phagocytosis and killing of coagulase-negative staphylococci, are impaired during long-term TPN. The impaired bactericidal activity seems to be related to defective intracellular killing in neutrophils. These findings may explain the high rate of septicemia caused by coagulase-negative staphylococci in infants receiving TPN. PMID- 10674622 TI - Assessment of telemedicine in surgical education and patient care. AB - OBJECTIVE: To analyze the value of teleconferencing for patient care and surgical education by assessing the activity of an international academic network. SUMMARY BACKGROUND DATA: The uses of telemedicine include teleeducation, training, and consulting, and surgical teams are now involved, sharing diagnostic information and opinions without the need for travel. However, the value of telematics in surgery remains to be assessed. METHODS: During a 2-year period, weekly surgical teleconferences were held among six university hospitals in four European countries. To assess the accuracy of telediagnosis for surgical cases, 60 randomly selected cases were analyzed by a panel of surgeons. Participants' opinions were analyzed by questionnaire. RESULTS: Seventy teleconferences (50 lectures and 271 case presentations) were held. Ninety-five of the 114 participants (83.3%) completed the final questionnaire. Eighty-six percent rated the surgical activity as good or excellent, 75.7% rated the scientific level as good or excellent, 55.8% rated the daily clinical activity as good or excellent, and 28.4% rated the manual surgical technique as good or excellent. The target organ was identified in all the cases; the organ structure and pathology were considered well defined in 93.3%, and the fine structure was considered well defined in 58.3%. Diagnosis was accurate in 17 cases (28.3%), probable in 25 (41.7%), possible but uncertain in 16 (26.7%), and not possible in 2 cases (3.3%). Discussion among the remote sites increased the rate of valuable therapeutic advice from 55% of cases before the discussion to 95% after the discussion. Eighty-six percent of the surgeons expressed satisfaction with telematics for medical education and patient care. CONCLUSIONS: Participant satisfaction was high, transmission of clinical documents was accurate, and the opportunity to discuss case documentation and management significantly improved diagnostic potential, resulting in an accuracy rate of up to 95%. Teleeducation and teleconsultation in surgery appear to be beneficial. PMID- 10674623 TI - Neurofibrillary tangles in progressive supranuclear palsy brains exhibit immunoreactivity to frameshift mutant ubiquitin-B protein. AB - In Alzheimer's disease (AD) neurofibrillary tangles (NFT) are strongly tau and ubiquitin immunopositive, and contain an aberrant form of ubiquitin derived from the ubiquitin-B gene denoted as UBB+1. We explored whether the tau-related NFT seen in another neurodegenerative disease, progressive supranuclear palsy (PSP), also showed an accumulation of UBB+1. Three cases of PSP were examined immunohistochemically for tau protein, ubiquitin-protein conjugates and UBB+1 using single and double labelling. We conclude that UBB+1 is associated with compact globose tangles rather than dispersed accumulations of tau in PSP, showing that its presence is not unique to AD. We propose that aggregation of ubiquitinated proteins into compact inclusions in PSP might be due to inhibition of the degradation of multiubiquitinated proteins by ubiquitin chains containing proximal UBB+1 rather than normal ubiquitin. PMID- 10674624 TI - Cyclosporine-A enhances choline acetyltransferase immunoreactivity in the septal region of adult rats. AB - Cyclosporine-A (CsA) is the primary anti-rejection drug used for organ and neural transplantation therapy. In addition to its immunosuppressive action, CsA has been recently shown to exert neuroprotective and neurotrophic effects in the central nervous system when able to cross the blood-brain barrier. Postulated mechanisms for these CsA-induced beneficial effects include the drug's powerful inhibition of the calcium-dependent phosphatase calcineurin (CN) and blockade of the assembly of the mitochondrial permeability transition pore. We report here, for the first time, that adult Wistar rats treated with CsA (10 mg/kg per day, i.p. for 9 days) displayed significantly reduced septal CN expression in combination with enhanced levels of septal choline acetyltransferase (ChAT) immunoreactivity as compared to controls. The observed enhancement of septal ChAT immunoreactivity suggests potential therapeutic utility of CsA for brain disorders characterized by alterations of the cholinergic system. PMID- 10674625 TI - Are there anticipatory segmental adjustments associated with lower limb flexions when balance is poor in humans? AB - For a leg raising task performed in a sagittal plane, it has been shown that body balance instability can suppress anticipatory postural adjustments (APAs). The aim of this study was to determine whether the global (centre of mass) postural adjustments were replaced by local (segmental) ones, which were compensating each other and resulting in a lack of global APAs. Six healthy subjects must perform a lower limb flexion from two initial postures, corresponding to a bipedal (Fbu) and an unipedal (Fuu) stance. Kinematics of postural adjustments were recorded with accelerometers. The results showed that in Fbu the kinematics have large durations of APAs, contrary to Fuu where there are none. They showed also that during the voluntary movement the magnitudes of the segmental postural accelerations were equal or superior in Fuu than in Fbu on the anteroposterior and lateral axes, where balance is poor. Also while, on the contrary, the magnitudes are reversed on the vertical axis. These results suggest that firstly: (1) the absence of APAs can correspond to a strategic response for weak postural base configuration and secondly; (2) the local postural accelerations, depending to the axes, are linked to two different functions: to maintaining the balance and to performing the focal movement. PMID- 10674626 TI - Structural comparison of zebrafish Elav/Hu and their differential expressions during neurogenesis. AB - The present communication reports the isolation and characterization of three new zebrafish elav/Hu (Kim, C.-H., Ueshima, E., Muraoka, O., Tanaka, H., Yeo, S.-Y., Huh, T.-L. and Miki, N., Zebrafish elav/HuC homologue as a very early neuronal marker. Neurosci. Lett., 216 (1996) 109-112) homologues, HuA, HuD and HuG. While HuA and HuG showed weak and ubiquitous expressions, HuD, as well as HuC, were specifically expressed in the neuronal cells. The first expression of HuD was detectable of the 10-somite stage, that is, several hours later than HuC. After 24 h of embryonic development, although HuD and HuC expressions overlapped overall, the cells expressing HuD were restricted to subsets of the HuC-positive neuronal cells in the brain and spinal cord. These differentially regulated spatial and temporal expression patterns implied distinct roles for HuC and HuD in neuronal determination and neuronal differentiation, respectively. PMID- 10674627 TI - Apoptotic cell death in rat brain following deltamethrin treatment. AB - In the present study we identified the degeneration and apoptotic cell death in rat brain after deltamethrin treatment. By hematoxylin-eosin (H&E) staining, a large number of degenerative cells (pyknosis of nuclei, disruption of eosinophilic cytoplasm) were seen in the hippocampus and cortex at 24 and 48 h after deltamethrin treatment at a dose of 12.5 mg/kg (i.p.) in corn oil. The similar morphological changes of degenerative cells were observed by cresyl violet staining. Numerous apoptotic cells were detected by in situ end labeling (ISEL) and flow cytometric analysis in the hippocampus and cortex at 24 and 48 h following deltamethrin treatment at the same dose, whereas no ISEL-positive cells were seen in the same brain regions of control rats. Moreover, using DNA gel electrophoresis, it was demonstrated that DNA fragmentation was markedly induced in the hippocampus and cortex at 24, 48 and 72 h after treatment. In addition, the protein synthesis inhibitor cycloheximide inhibited the DNA fragmentation elicited by deltamethrin in rat brain. These results indicate that deltamethrin induces degeneration and apoptotic cell death in rat brain, suggesting an important role played by apoptosis in neurotoxicity of deltamethrin. PMID- 10674628 TI - Immunohistochemical localization of neurocalcin in human sensory neurons and mechanoreceptors. AB - The localization of neurocalcin in the developing and adult human peripheral nervous system (dorsal root and sympathetic ganglia (DRG, SG), and enteric nervous system (ENS)) was investigated using immunohistochemistry. A subpopulation of large-sized neurons in DRG of 9 and 12 weeks old embryos showed immunoreactivity (IR), whereas the sympathetic ganglia or enteric neurons did not. In adults, neurocalcin IR was restricted to a subpopulation of large (13%) and intermediate (15%) sized neurons in DRG. The protein was also found in muscular (67%) and cutaneous (12%) nerve fibers, as well as in the axons supplying muscular (muscle spindles, Golgi's tendon organs, and perimysial Pacinian corpuscles) and cutaneous (Meissner's but not Pacinian corpuscles) mechanoreceptors, as well as motor end-plates. Present results demonstrate that neurocalcin in both developing and adult humans can be used as a specific marker for a subpopulation of sensory neurons coupled to proprioception and touch, and for axons of motoneurons forming motor end-plates. PMID- 10674629 TI - Argatroban, a thrombin inhibitor, decreased mortality after 10 min of forebrain ischemia in the gerbil. AB - We investigated whether anticoagulant therapy with heparin or a selective thrombin inhibitor, argatroban, may ameliorate the postischemic cerebral circulation and attenuate mortality after 10 min of forebrain ischemia. Postischemic subcutaneous injection of argatroban (5 mg/kg) significantly attenuated mortality (9.1%) compared with non-treatment (45.5%) during 14 days' observation period. This effect coincided with: (1) increased cortical CBF after reperfusion; (2) attenuation of brain edema; and (3) less severe cell damages in the cerebral cortex. In contrast, nine of the 22 gerbils treated with heparin (830 IU/kg) were found dead on the next day due to massive bleeding in the surgical wound and 13 bleeding-avoided gerbils did not show significant amelioration in mortality (30.8%). These findings suggest that argatroban is an effective anticoagulant for prevention of cell damage after a relatively long forebrain ischemia. PMID- 10674630 TI - Activation of calpain precedes morphological alterations during hydrogen peroxide induced apoptosis in neuronally differentiated mouse embryonal carcinoma P19 cell line. AB - In order to reveal neurodegeneration elicited by reactive oxygen intermediates (ROI), neuronally differentiated cells from mouse embryonal carcinoma P19 cell line were exposed to hydrogen peroxide (H2O2). Enhanced protein-phosphorylation on tyrosine residues was detectable within 5 min of exposure to H2O2, and gradual rises in intracellular free Ca2+ level and in calpain activity were observed. Furthermore, H2O2 stimulation of differentiated P19 cells for 24 h resulted in morphological alterations in somas as well as neurites. Also, within 6 h of H2O2 treatment DNA fragmentation has been detected. Taken together, these results suggest that oxidative stress induces degradation of cytoskeletal proteins presumably resulting from increased intracellular Ca2+ concentration and subsequent activation of calpain. PMID- 10674631 TI - Expression of induced nitric oxide synthase in amoeboid microglia in postnatal rats following an exposure to hypoxia. AB - The present study showed the expression of induced nitric oxide synthase (iNOS) immunoreactivity in amoeboid microglia following an exposure to transient hypoxia in postnatal rats. iNOS immunoreactivity was expressed mainly in the amoeboid microglia in corpus callosum and subependymal regions of the ventricles within 3 h after hypoxia. The expression declined after 5 h, and became undetectable after 15 h and in longer surviving rats. The immunoreactivity of these cells with OX 42, which is a marker for microglia cells and detects complement type three receptors (CR3), was comparable in the rats exposed to hypoxia and the control rats. Immunoglobulin G (IgG) immunoreactivity was observed in the amoeboid microglia up to 3 h after hypoxia but it was undetectable in longer surviving rats and in the control rats. The iNOS expression in the amoeboid mircoglial cells may be related to the host defense and maintenance of structural integrity of the highly vulnerable periventricular white matter after hypoxia. The immunostaining of amoeboid microglial cells with IgG following hypoxia indicates leakage of plasma immunoglobulin from the blood vessels and its removal by the amoeboid microglial cells. PMID- 10674632 TI - Chronic hyperglycaemia increases neuronal sensitivity to high potassium in hippocampal slices from streptozotocin-treated diabetic rats. AB - The chronic hyperglycaemia associated with diabetes mellitus increases susceptibility to epileptiform-like activity in the central nervous system. Changes in extracellular potassium levels directly influence neuronal excitability and significantly one of the major regulators of extracellular potassium, the Na-K ATPase, is known to be down-regulated by chronic hyperglycaemia. The sensitivity of hippocampal slices, from rats made diabetic for 2-3 weeks with streptozotocin, to increases in the extracellular potassium concentration ([K+]o) was, therefore, investigated. Raising [K+]o to 10 mM increased the number of orthodromically-evoked population spikes (PSs) in area CA1. This recruitment was significantly greater in hippocampal slices from diabetic rats, which also exhibited significantly more spontaneous activity. These findings confirm a diabetes-dependent increase in sensitivity of central neurones to changes in [K+]o and may help to explain the increased susceptibility to epileptiform activity in this disease state. PMID- 10674633 TI - Diurnal variation in orexin A immunoreactivity and prepro-orexin mRNA in the rat central nervous system. AB - Orexins are a family of neuropeptides originally believed to be important mediators of food intake. The wide distribution of orexins and their receptors, however, has suggested other regulatory functions for these peptides including involvement in sleep and arousal mechanisms. In this study, we have demonstrated diurnal variation in orexin A immunoreactivity in the pons, from where locus coeruleus noradrenergic neurones innervate other brain areas to stimulate arousal, and in the preoptic/anterior hypothalamic region, an area implicated in the regulation of sleep and circadian rhythms. Orexin A immunoreactivity decreased by 50% in the preoptic/anterior hypothalamus from 09:00 to 21:00 h (P < 0.0001), whilst in the pons, it increased by over 30% from 09:00 to 01:00 h (P = 0.02). Prepro-orexin mRNA also displayed diurnal variation. This further suggests that orexins are involved in the regulation of the sleep/wake cycle. PMID- 10674634 TI - Receptor mechanisms mediating differentiation and proliferation effects of retinoids on neuroblastoma cells. AB - The aim of this study was to clarify retinoid receptor mechanisms mediating the effects of 9-cis retinoic acid (RA) and investigate the ability of RAR- and RXR specific analogues to induce differentiation and inhibit proliferation in neuroblastoma cells. Differentiation and the inhibition of proliferation by 9-cis RA, but not all-trans RA, were inhibited by the RXR-homodimer antagonist LG745. The RXR-specific agonist LGD1069 was ineffective at inducing differentiation or inhibiting proliferation, but showed marked synergism with RAR-specific agonists with respect to inhibiting proliferation. These data suggest that the effects of 9-cis RA are mediated via both RXR-homodimers and heterodimers. However, combinations of RAR- and RXR-selective analogues were not as effective at promoting differentiation. This study indicates that different receptor mechanisms are involved in retinoid-induced differentiation and inhibition of proliferation in neuroblastoma cells. PMID- 10674635 TI - Age-dependent changes in phosphorylated cAMP response element-binding protein immunoreactivity in motoneurons of the spinal nucleus of the bulbocavernosus of male rats. AB - Phosphorylated cAMP response element-binding protein (CREB) immunoreactivity was examined in motoneurons of the spinal nucleus of the bulbocavernosus (SNB) in young and old male rats by immunohistochemistry. In young animals, intense CREB immunoreactivity was confined to the cell nucleus, but not in the nucleolus of SNB motoneurons. In old animals, both the intensity of CREB immunoreactivity in the nuclei and number of CREB immunoreactive nuclei of the SNB motoneurons were significantly reduced. A marked decline in expression of CREB in the aged SNB motoneurons suggests alternation of cAMP-mediated regulation of gene expression in the SNB system with advancing age. PMID- 10674636 TI - Endogenous adenosine regulates the effects of low-frequency stimulation on the induction of long-term potentiation in CA1 neurons of guinea pig hippocampal slices. AB - A train of low-frequency afferent stimuli (LFS, 1 Hz, 1000 pulses), given 60 min prior to a tetanus (100 Hz, 100 pulses), suppresses the induction of long-term potentiation (LTP) in which a short-term potentiation decreases gradually back to the pre-tetanic level within 40-50 min (LTP suppression). We investigated the effects of adenosine A1 or A2 receptor antagonists (8-cyclopentyltheophylline (8 CPT) and CP-66713, respectively) on LTP suppression in CA1 neurons of guinea pig hippocampal slices. When the LFS was delivered in the presence of 8-CPT (1 microM), LTP suppression was not significantly affected. However, when LFS was delivered in the presence of CP-66713 (10 microM), LTP suppression was inhibited, leading to successful LTP induction. These results indicate that endogenous adenosine, acting via A2 receptors, is involved in the mechanism of LTP suppression. PMID- 10674637 TI - Synapsin III gene polymorphisms and schizophrenia. AB - Synapsins are synaptic vesicle-associated phosphoproteins and are thought to play crucial roles in synaptogenesis and neurotransmitter release. Synaptic abnormalities have been reported in the pathophysiology of schizophrenia. In addition, the synapsin III gene, a member of the synapsin gene family, has been located at 22q12-13, which has been suggested as a potential susceptibility locus for schizophrenia. We investigated a genetic association between schizophrenia and the synapsin III gene polymorphisms (-631C/G and -196G/A) in 160 schizophrenic patients and 153 controls. No significant positive association between either polymorphism and schizophrenia was observed. Furthermore, no significant association was observed between either polymorphism and the diagnostic subtypes. Our results suggested that the synapsin III gene polymorphisms do not confer increased susceptibility to schizophrenia. PMID- 10674638 TI - Melioidosis as an emerging global problem. AB - There is remarkably little known about the incidence of melioidosis outside a few countries (Thailand, Australia, Singapore and Malaysia). Presumably it is widespread in tropical south east Asia. Elsewhere there are tantalising glimpses of the tip of what may be a large iceberg. Since a specific diagnosis of melioidosis requires awareness on the part of clinicians, and the existence of a laboratory capable of isolating and identifying Burkholderia pseudomallei, a luxury not available in most rural tropical areas, the size of this iceberg is likely to remain unknown for the foreseeable future. There is mounting evidence that the disease is endemic in the Indian sub-continent and the Caribbean, and there have been unsubstantiated reports of recent cases in South Africa and the Middle East. It is unclear whether melioidosis has really spread to such areas relatively recently, or has been there but unrecognised for a long time. Almost all cases diagnosed in temperate climates have been imported from the tropics, with the exception of a unique outbreak which occurred in France in the mid 1970s. With increasing world wide travel of both humans and other animals, the potential exists for melioidosis to spread to new and fertile pastures. PMID- 10674639 TI - The epidemiology of melioidosis in Australia and Papua New Guinea. AB - Melioidosis was first described in Australia in an outbreak in sheep in 1949 in north Queensland (22 degrees S). Human melioidosis was first described from Townsville (19 degrees S) in 1950. Melioidosis is hyperendemic in the Top End of the Northern Territory (NT) and as in parts of northeastern Thailand it is the commonest cause of fatal community-acquired septicemic pneumonia. In the 9 years since 1989 the prospective NT melioidosis study at Royal Darwin Hospital (12 degrees S) has documented 206 culture confirmed cases of melioidosis, with an average annual incidence of 16.5/100,000. Melioidosis is also seen in the north of Western Australia and north Queensland, including the Torres Strait Islands, but is uncommon in adjacent Papua New Guinea. Serological studies suggest that infection is rare in the Port Moresby region, but there is emerging evidence of melioidosis from Western Province. The NT study has documented inoculating events in 52 (25%) of cases, with an incubation period of 1-21 days (mean 9 days); 84% of cases had acute disease from presumed recent acquisition and 13% had chronic disease (sick, > 2 months). In 4% there was evidence of possible reactivation from a latent focus; 28 of 153 (18%) males had prostatic abscesses. The overall mortality was 21% (43 cases), with a mortality rate in septicemic cases (95) of 39% and in non-septicemic cases (103) of 4%. Pneumonia was the commonest presentation in both groups and, in addition, eight patients (two deaths) presented with melioidosis encephalomyelitis. Melioidosis clusters in temperate Australia are attributed to animals imported from the north. Molecular typing of Burkholderia pseudomallei isolates from temperate southwest Western Australia showed clonality over 25 years. In this outbreak and in studies from the NT, some soil isolates are molecularly identical to epidemiologically related animal and human isolates. Molecular typing has implicated the water supply in two clonal outbreaks in remote aboriginal communities in northern Australia. Further prospective collaborative studies are required to evaluate whether there are truly regional differences in clinical features of melioidosis and to better understand how B. pseudomallei is acquired from the environment. PMID- 10674640 TI - Melioidosis in Southeast Asia. PMID- 10674641 TI - Recent advances in the treatment of severe melioidosis. AB - For the last decade, high-dose intravenous ceftazidime has been the drug of choice for the treatment of severe melioidosis, after ceftazidime was shown to be superior to the 'conventional' four-drug regimen (chloramphenicol, doxycycline and trimethoprim-sulphamethoxazole) in a randomised trial. Combination ceftazidime-trimethoprim-sulphamethoxazole was compared with the conventional regimen in a separate trial with similar results, but we still do not know whether such combination therapy is needed in melioidosis. Co-amoxiclav (amoxycillin-clavulanate) has been shown to be effective but was associated with a higher rate of treatment failure than ceftazidime. Two further treatment trials in acute melioidosis have recently been conducted in Thailand. In the first of these, high-dose intravenous imipenem was compared with ceftazidime and the results suggest that the two regimens possess similar efficacy. Cefoperazone sulbactam has been compared with ceftazidime-trimethoprim-sulphamethoxazole in a small number of patients, and further results are awaited. Relapses of melioidosis should be treated in a similar manner to primary infections. PMID- 10674642 TI - Antimicrobial resistance in Burkholderia pseudomallei. AB - Four strains of Burkholderia pseudomallei were used to determine the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time kill curves with 13 single antimicrobial agents: ceftazidime, piperacillin, imipenem, amoxicillin/clavulanic acid, doxycycline, cotrimoxazole, kanamycin, rifampicin, ciprofloxacin, trovafloxacin, clarithromycin, azithromycin and meropenem. The time-kill studies were also performed with 33 pairs of combinations of the above antimicrobial agents: 15 combinations which would be expected to be used for acute therapy and 18 combinations for maintenance therapy. The results show that the single and combination antimicrobial agents with bactericidal effects against the four strains of B. pseudomallei which should be used for clinical trials in acute melioidosis are: imipenem, meropenem, and imipenem + azithromycin. The combination antimicrobial agents which should be further studied for the ability to eliminate biofilm and intracellular killing effect are ciprofloxacin + clarithromycin, ciprofloxacin + azithromycin, and imipenem + azithromycin. PMID- 10674643 TI - Neurological melioidosis. AB - Neurological abnormalities have long been recognised in animals with melioidosis, including laboratory rodents and sheep in the first Australian outbreak in 1949. Autopsies in animals have shown microabscesses and lymphocytic infiltration to be present on occasion in the same animal, but Burkholderia pseudomallei is usually able to be grown from central nervous system (CNS) tissue. In humans CNS melioidosis is unusual, but both macroscopic brain abscesses and encephalitis occur. There has been a recently recognised syndrome of meningoencephalitis with varying involvement of brainstem, cerebellum and spinal cord. The prospective melioidosis study at Royal Darwin Hospital has documented 12 cases of CNS melioidosis over 9 years out of a total of 232 cases of melioidosis (5%). Prominent features on presentation were unilateral limb weakness (6), predominant cerebellar signs (2), mixed cerebellar and brainstem features with peripheral weakness (2) and flaccid paraparesis (2). Eight patients had unilateral VIIth nerve palsy and six bulbar palsy, with five requiring prolonged ventilation. Brain CT scans are usually normal initially, but MRI shows dramatic changes. Three patients died and only three made a full recovery. In two patients with predominant mononuclear CSF pleocytosis, B. pseudomallei was cultured from CSF and autopsy in one of these showed necrotising encephalitis with microabscesses. Although it has been postulated that a neurotropic exotoxin may account for melioidosis encephalomyelitis, the recent findings and comparison with the animal data suggest that direct organism spread within the CNS may be primarily responsible. Preliminary molecular typing of isolates shows no evidence of a specific strain of B. pseudomallei responsible for CNS melioidosis end further studies are required to determine if the apparent higher rate of CNS disease in Australia is due to true regional differences or is from increased ascertainment. PMID- 10674644 TI - Animal melioidosis in Australia. AB - Melioidosis was first diagnosed in Australia in sheep in 1949. While it has been considered endemic in tropical Australia, there have been animal outbreaks in southwest Western Australia and southern Queensland. Infection occurs in many species, with both latency and a wide range of clinical manifestations. Some species may develop melioidosis only if immunocompromised. Sheep and goats are particularly susceptible, resulting in the requirement for pasteurisation of tropical commercial goat's milk. Nine out of 43 (21%) goats had aortic lesions at autopsy and seven died from aortic aneurysm rupture. Transplacental transmission in goats has also been documented. Asymptomatic organ abscesses are common in pigs but bovine melioidosis is very rare. Camels moved north and an alpaca brought to Darwin have died from melioidosis. It also occurs in wildlife, including birds, crocodiles and kangaroos. Zoonotic transmission to humans is extremely unusual, but there are many similar epidemiological and clinical features of melioidosis in animals and humans. There have been three possible zoonotic cases in Australia. Molecular typing has found identical Burkholderia pseudomallei organisms from animals, humans and soil. The study of melioidosis in animals, especially the use of molecular genetic techniques for organism identification and typing, will continue to unravel aspects of the disease that remain unclear in humans. PMID- 10674645 TI - Ecology of Burkholderia pseudomallei and the interactions between environmental Burkholderia spp. and human-animal hosts. AB - Early workers thought that melioidosis was a zoonosis with a reservoir in rodents, but we now know that Burkholderia pseudomallei is a widely distributed environmental saprophyte. In northeast Thailand, two thirds of paddy fields yield the organism, and 80% of children have antibodies by the time they are 4 years old. However, interpretation of these results has been complicated by the recent recognition of avirulent, antigenically cross-reacting environmental organisms for which the name B. thailandensis has been proposed. We still know very little about the climatic, physical, chemical and biological factors which control the proliferation and survival of Burkholderia spp. in the environment, although epidemiological studies show space-time clustering of melioidosis. It is assumed that most human and animal melioidosis arises through exposure to contaminated soil or muddy water, although only 6% of human cases have a clear history of inoculation, and a further 0.5% of cases follow near-drowning. Laboratory animals have also been infected by ingestion, inhalation and insect bites, but evidence of infection acquired naturally by these routes remains anecdotal. Sporadic cases have resulted from iatrogenic inoculation, laboratory accidents, and person-to person or animal-to-person spread. Whether exposure to B. pseudomallei will result in disease probably depends on the balance between the virulence of the strain, the immune status of the host (e.g. diabetes mellitus) and the size of the inoculum. PMID- 10674646 TI - Multiple replicons constitute the 6.5-megabase genome of Burkholderia pseudomallei. AB - Burkholderia pseudomallei is a causative agent of melioidosis, a fatal tropical infectious disease endemic in Southeast Asia and Northern Australia. In order to determine the size and characteristics of the bacterial genome, the B. pseudomallei genome and genes were analyzed by pulsed field gel electrophoresis of the undigested, intact megabase DNA, and by computational analysis of nucleotide sequences of B. pseudomallei genes which have been sequenced by several investigators and already deposited in a public database. The results showed that the B. pseudomallei genome consists of two large replicons, and that both contain ribosomal RNA gene sequences, indicating the presence of two chromosomes. The classical arabinose-negative B. pseudomallei isolate K96243 has chromosomes of approximately 3563 +/- 73 and 2974 +/- 40 kilobase-pairs in size, giving a total genome size of about 6.5 million base-pairs. The arabinose positive nonvirulent biotype of B. pseudomallei also has two replicons which are smaller than those of the arabinose-negative biotype. Analysis of the publicly available nucleotide sequences showed that the average B. pseudomallei gene is approximately 1031 base-pairs in size, with an average G + C content of 65.7%. The genome is gene-rich and about 89% of the coding capacity is used as coding sequences. It can therefore be estimated that the entire B. pseudomallei genome encodes about 5600 genes. PMID- 10674647 TI - Molecular phylogeny of Burkholderia pseudomallei. AB - In terms of population structure, the species Burkholderia pseudomallei contains both clonal and non-clonal elements. By indexing variation in rRNA loci using the restriction endonuclease BamHI, we found that two ribotypes (types 1 and 3) are predominant in nature. Ribotype 3 is prevalent in Asian countries while ribotype 1 is more widespread. Some disease association was suggested for 4 ribotypes and strains of ribotype 4 were markedly associated with a fatal outcome. DNA macrorestriction (XbaI) profiles resolved by pulsed-field gel electrophoresis revealed great heterogeneity within the prevalent ribotypes and these profiles appeared to be reliable strain markers. Arabinose environmental strains were characterised by BamHI ribotypes that were markedly distinct form clinical and environmental isolates of the arabinose negative phenotype. PMID- 10674648 TI - Genome fingerprinting by pulsed-field gel electrophoresis of isolates of Burkholderia pseudomallei from patients with melioidosis in Thailand. AB - A total of 95 isolates of Burkholderia pseudomallei from 53 sporadic cases in Thailand were examined by pulsed-field gel electrophoresis. Digestion of genomic DNA of all isolates by NcoI generated a macrorestriction pattern similar to that of B. pseudomallei which cannot assimilate L-arabinose. Analysis using restriction enzymes SpeI and AvrII demonstrated greater sensitivity than NcoI digestion in the differentiation of B. pseudomallei and could be used for epidemiological groupings. Four cluster groups were evident among 37 isolates tested and the majority of isolates within each cluster displayed more than 65% similarity. Furthermore, multiple isolates from 18 and 35 patients with single and recurrent episodes of melioidosis, respectively, were examined. All patients with a single episode yielded genetically identical isolates and four of 35 patients with recurrent episodes were infected with strains of different genotypic patterns from the primary isolate(s). Hence, most repeated episodes of infection in melioidosis are as a result of the original infecting strains. PMID- 10674649 TI - Use of multiplex PCR patterns as genetic markers for Burkholderia pseudomallei. AB - A simple PCR-based typing method was developed to differentiate between strains of Burkholderia pseudomallei. Two pairs of primers, based on sequences from two specific DNA probes, were used to amplify the bacterial DNA by multiplex PCR. We evaluated the PCR method for epidemiological typing of B. pseudomallei and compared this with restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) methods. In RFLP, the DNA of B. pseudomallei was digested with HindIII and the pKKU-S23L was used as a probe while 5' GTTTCGCTCC 3' primer was used in RAPD. DNA was obtained from 37 B. pseudomallei environmental and clinical isolates from humans or animals. These isolates were also classified by their ability to assimilate L-arabinose. A total of 21 type patterns were identified by multiplex PCR. Among human and animal isolates, multiplex PCR revealed ten types, all of which were arabinose negative (Ara-), whereas six of the 11 types of environmental isolates were Ara-. There are two environmental patterns that also were found in clinical isolates. The RFLP technique showed 12 different types in the 37 isolates, and three of these contained both Ara+ and Ara- isolates. The RAPD technique revealed 33 different types in the 37 isolates. Multiplex PCR, therefore, is the genetic marker that best correlates with the ability of the organism to assimilate L-arabinose. Moreover, two types (M4, M15) correlated with disseminated septicemic melioidosis in the northeast Thailand. If a greater number of isolates are tested, the multiplex PCR technique may prove to be useful for rapid epidemiological typing of B. pseudomallei. PMID- 10674650 TI - Pathogenesis of and immunity to melioidosis. AB - While Burkholderia pseudomallei, the causative agent of melioidosis, is becoming increasingly recognized as a significant cause of morbidity and mortality in regions to which it is endemic, no licensed vaccine preparation currently exists for immunization against the disease. Therefore, one of the primary goals of our research has been to identify and characterize antigens expressed by B. pseudomallei isolates for the intended purpose of developing a vaccine construct that can be used to actively immunize specific high risk populations against the disease. By utilizing a combination of biochemical, immunological and molecular approaches, our studies now indicate that some of the most promising candidates for this task include flagellin proteins and the endotoxin derived O polysaccharide (PS) antigens expressed by the organism. In this review, we have attempted to summarize the current status of B. pseudomallei research while endeavoring to provide a rationale for our approach towards the development of a melioidosis vaccine. PMID- 10674651 TI - Exopolysaccharides of Burkholderia pseudomallei. PMID- 10674652 TI - Protease production by Burkholderia pseudomallei and virulence in mice. AB - The aim of this study was to assess protease production and virulence of various Burkholderia pseudomallei strains. Protease activity was evaluated in filtrates from cultures grown for 50 h in TSB Dialysate by azocasein hydrolysis, and expressed as absorbancy at 405 nm. Virulence was assessed in 8 weeks old SWISS mice, by intraperitoneal injection of 6-6 x 10(5) CFU, and the LD50 was calculated after 30 days by the method of Reed and Muench. The lethal activity was studied for five strains of B. pseudomallei and the type strains of Burkholderia pseudomallei, Burkholderia mallei, and Burkholderia cepacia. The three type strains appeared to be low protease producers (A405 = 0.11, 0.09 and 0.00, respectively) and avirulent. The two more virulent B. pseudomallei strains exhibited significantly different LD50, 3.5 x 10(2) (IPP 6068 VIR) versus 2.1 x 10(5) CFU/mouse (40/97), and protease activities (A405 = 0.046 and 0.79, respectively). Moreover, the avirulent parent of IPP 6068 (AG), was a better protease producer than the 6068 VIR strain, A405 = 0.26 versus 0.046. These results suggest that there is no correlation between virulence and level of exoproteolytic activity, when B. pseudomallei is injected to mice via the intraperitoneal route. PMID- 10674653 TI - Shedding of lipopolysaccharide and 200-kDa surface antigen during the in vitro growth of virulent Ara- and avirulent Ara+ Burkholderia pseudomallei. AB - Non-virulent Ara+ B. pseudomallei environmental isolates differ from virulent Ara clinical isolates by their ability to assimilate L-arabinose and the absence of a 200 kDa antigen on their surface. The latter, present only on the Ara- isolates from either clinical or environmental origin, was recently demonstrated by its immunoreactivity with monoclonal antibody (MAb) 5F8. We recently demonstrated that lipopolysaccharide (LPS) from both biotypes were indistinguishable from one another with regard to SDS-PAGE profiles and immunoreactivities with immune sera. In this study, the shedding of LPS and 200-kDa antigen into the culture medium during the in vitro growth of Ara- was compared with that of its Ara+ counterpart, using MAb-based sandwich ELISAs. The results showed that the LPS shedding profiles from the two biotypes were similar to one another. This was in contrast to the situation with the 5F8-reactive antigen. The culture fluid of all Ara- isolates and none of the Ara+ isolates were found to react strongly with the MAb 5F8 during the early log phase of growth. However, during the late stationary phase, a trace amount of the 5F8-reactive material could also be detected in the culture fluid of the Ara+ isolates. PMID- 10674654 TI - Proinflammatory cytokine mRNA responses in experimental Burkholderia pseudomallei infection in mice. AB - Melioidosis is a potentially fatal disease of both human and animals caused by the bacterium Burkholderia pseudomallei. Disease is endemic in tropical and subtropical regions of Southeast Asia and Northern Australia. The pathogenesis of melioidosis is poorly understood. In particular, the host responses that occur following infection, and the specific host-pathogen interactions that result in the development of either acute or chronic infection are unclear. Using an established murine model, we investigated early proinflammatory cytokine responses believed to be critical in the development of acute and chronic B. pseudomallei infection. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to assess levels of mRNA for tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta) and interleukin 6 (IL-6) in the liver of mice following infection. We demonstrate that the level of mRNA for these cytokines increase moderately in chronic infection in C57BL/6 mice. However, in acute infection in BALB/c mice, mRNA responses for these cytokines were shown to be comparatively greater. These results demonstrate that early proinflammatory cytokine responses are important in the immunopathogenesis of melioidosis. PMID- 10674655 TI - Recent developments in laboratory diagnosis of melioidosis. PMID- 10674656 TI - Comparison of three PCR primer sets for diagnosis of septicemic melioidosis. AB - Several sets of PCR primers have recently been developed for detection of Burkholderia pseudomallei. In this report, the performance of 16S rRNA gene primers (16S), rRNA spacer gene primers (spacer), and 'LPS' primers (LPS) were compared. All primer sets were tested by PCR amplification of the same DNA samples extracted from blood specimens of 46 patients from northeastern Thailand, of which 29 had melioidosis based on blood culture as a gold standard. The sensitivities were 41, 35.7, and 31% while the specificities were 47, 59, and 100% for the 16S, spacer, and LPS primers, respectively. The positive predictive values were 60, 59, and 100%, while negative predictive values were 35, 34, and 46%, for these primers. The low sensitivity of PCR was suspected to be because of small numbers of bacteria in the samples. In addition, one primer set could not detect all B. pseudomallei strains. To make PCR for melioidosis more practical, bacterial concentration steps must be added. Lastly, mixed infection of patients in endemic areas may be the cause of controversial false positive PCR results, and should be further investigated. PMID- 10674657 TI - Malaria gametocytes. PMID- 10674658 TI - Hantavirus (Bunyaviridae) infections in rodents from Orange and San Diego Counties, California. PMID- 10674659 TI - Hantavirus infections in rodents PMID- 10674660 TI - A phase I safety and immunogenicity trial with the candidate malaria vaccine RTS,S/SBAS2 in semi-immune adults in The Gambia. AB - RTS,S is a novel pre-erythrocytic malaria vaccine based on the circumsporozoite surface protein (CSP) of Plasmodium falciparum linked to hepatitis B surface antigen (HBs) and combined with a novel adjuvant system (SBAS2). We have conducted a Phase I trial with three doses of this vaccine given at 0, 1, and 6 months to 20 semi-immune, adult, male volunteers in The Gambia to assess its safety and immunogenicity. Eighteen of the 20 volunteers completed the study. There were no clinically significant local or systemic adverse events following each vaccination. Hematologic and biochemical indices before and two weeks after each vaccination showed no evidence of toxicity. Antibody titers to both CSP and HBs showed a significant increase after vaccination; these were the largest after the third dose. We conclude that the RTS,S/SBAS2 vaccine induces no significant toxicity in this semi-immune population and produces significant increases in antibody titers to CSP. PMID- 10674661 TI - Safety, immunogenicity, and lot stability of the whole cell/recombinant B subunit (WC/rCTB) cholera vaccine in Peruvian adults and children. AB - To assess the safety, immunogenicity, and lot stability of the whole cell/recombinant B subunit cholera vaccine, 2 lots manufactured in June 1991 and February 1992 were tested in January 1995. Two oral doses of vaccine or placebo given 2 weeks apart were given with buffer to 216 Peruvian adults and children. Symptoms were elicited for 3 days after each dose. Serum and plasma specimens obtained from each volunteer before vaccination and 10-14 days after the second dose were tested for vibriocidal and anti-cholera toxin antibodies. The vaccine was well-tolerated. Nearly half of the 100 vaccinees had pre-vaccination vibriocidal titers > or = 1:40. Elevated titers were observed in 22% of 37 children 2-5 years of age compared with 66% of 63 vaccinees 6-65 years (P < 0.001). A > or =2-fold serum vibriocidal response was observed in 55% of 100 vaccinees and 6% of 32 placebo recipients. An elevated pre-vaccination titer (< or =1:40) did not change the proportion of vaccinees who responded with a > or =2 fold increase in vibriocidal titer (51% versus 59%, difference not significant), but did change the proportion responding with a > or =4-fold increase (41% versus 22%; P < 0.05). The vibriocidal seroconversion rate was lowest in children 2-5 years old despite low pre-vaccination titers. Two-fold or greater serum antitoxic responses in IgA and IgG were observed in >90% of the vaccinees; > or =4-fold responses were seen in 65-70% of the vaccinees with a 6-8-fold increase over baseline. Plasma specimens were as good as sera for determining anti-toxic antibodies by ELISA, but were less satisfactory for determining vibriocidal antibody titers. PMID- 10674662 TI - An accelerated schedule for tick-borne encephalitis vaccine: the American Military experience in Bosnia. AB - Tick-borne encephalitis (TBE) is a viral illness endemic to the Balkan region. United States military forces were deployed to Bosnia in early 1996 as part of Operation Joint Endeavor, a U.S.-led multinational peace-keeping operation. To counteract the TBE threat, an inactivated, parenteral vaccine (FSME-Immun Inject; Immuno AG, Vienna, Austria) was offered to soldiers at high risk on a volunteer basis in an accelerated, 3-dose schedule (0, 7, and 28 days). Passive adverse reaction surveillance was conducted on 3,981 vaccinated personnel. Paired sera from a randomly selected group of 1,913 deployed personnel (954 who received vaccine and 959 who were unvaccinated) were tested for antibodies to TBE by an ELISA. Three-dose recipients demonstrated an 80% seroconversion rate (4-fold or greater increase in anti-TBE titers). By comparison, the TBE infection rate in the unvaccinated cohort was found to be only 0.42% (4 of 959). Only 0.18% of vaccinees reported self-limited symptoms. An accelerated immunization schedule appears to be an acceptable option for military personnel or travelers on short term notice to TBE-endemic areas. PMID- 10674663 TI - Rabies mass vaccination campaigns in Tunisia: are vaccinated dogs correctly immunized? AB - Among the 301 dogs vaccinated against rabies in a suburban area of Tunis, 165 were sero-surveyed for 13 months. One month after vaccination, 65% of the dogs less than 1 year of age and 76-84% of the older dogs had significant antibody titers. One month after annual revaccination, these percentages ranged between 92% and 100%. Puppies (less than 3 months old) responded to vaccination with no significant interference by passive maternal immunity. Based on these percentages, a 93% rate of protection may be expected for vaccinated dogs. This study confirms that all dogs (even those less than 3 months of age) must be vaccinated during mass campaigns. The expected protection conferred by locally produced potent vaccines reaches 79-99% based on the age of the dogs. The alleged relative inability of local dogs to respond to vaccination cannot explain the partial success of rabies control in Tunisia. PMID- 10674664 TI - An outbreak of West Nile fever among migrants in Kisangani, Democratic Republic of Congo. AB - In February 1998, an outbreak of acute febrile illness was reported from the Kapalata military camp in Kisangani, the Democratic Republic of Congo. The illness was characterized by an acute onset of fever associated with severe headache, arthralgia, backache, neurologic signs, abdominal pain, and coughing. In 1 individual, hemorrhagic manifestations were observed. The neurologic signs included an altered level of consciousness, convulsions, and coma. Malaria was initially suspected, but the patients showed negative blood films and failed to respond to antimicrobial drugs. A total of 35 sera collected from the military patients in the acute phase were tested for the presence of IgM against vector borne agents. Serum IgM antibodies against West Nile fever virus were found in 23 patients (66%), against Chikungunya virus in 12 patients (34%), against dengue virus in 1 patient (3%), and against Rickettsia typhi in 1 patient (3%). All sera were negative for IgM antibody against Rift Valley fever virus, Crimean Congo hemorrhagic fever virus, and Sindbis virus. These data suggest that infections with West Nile fever virus have been the main cause of the outbreak. PMID- 10674665 TI - Hepatitis C: prevalence and common genotypes among ethnic groups in Jeddah, Saudi Arabia. AB - The presence of antibodies to hepatitis C virus (HCV) was examined using a third generation ELISA in 3,350 serum samples obtained from patients visiting different outpatient clinics, preoperative patients, and women visiting for ante-natal care at the Dr. Fakeeh hospital in Jeddah, Saudi Arabia from January to September 1998. The population included mainly Saudis, Egyptians, and Yemenis, and patients from other Middle East and Asian countries. The prevalence of HCV infection was 5.87% among Saudis, 22.54% among Egyptians, and 2.12% among Yemenis. The prevalence among patients from other Middle East and Asian countries were 3.38% and 4.98%, respectively. The overall prevalence among the studied group was 6.75%. Genotyping of isolates from Saudi Arabia showed that the prevalences of types 4, la, 1b, and 2 were 45.9%, 40.6%, 10.85%, and 2.7%, respectively. Genotyping of isolates from Egypt showed that type 4 was the major type among Egyptians (91.6%), while types 1a and 1b each had prevalences of 4.2%. PMID- 10674666 TI - Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua. AB - This report presents the results of applying the reverse transcriptase-polymerase chain reaction (RT-PCR) to the analysis of clinical specimens during the 1998 dengue epidemic in Nicaragua. The RT-PCR was validated through comparison with viral isolation, resulting in a sensitivity of 100% and a specificity of 90%. In country application of the RT-PCR permitted the rapid identification of dengue-3 virus as the cause of the epidemic at the beginning of 1998 and the detection of the reintroduction of dengue-2 virus in the middle of the year. Nineteen isolates of dengue-3 and one of dengue-2 were characterized using the restriction site specific (RSS)-PCR technique. This showed that the dengue-3 strain belonged to the "Sri Lanka" subtype and that the dengue-2 strain belonged to the "Jamaica" subtype, both of which have been associated with hemorrhagic dengue in the Americas. The application of these simple PCR-based strain typing methods in a country endemic for dengue virus infections can help to characterize the transmission dynamics of this important emerging infectious disease problem and provide this information to local health authorities in a timely manner so that appropriate control measures can be implemented. PMID- 10674667 TI - Shipboard impact of a probable Norwalk virus outbreak from coastal Japan. AB - Norwalk virus has been implicated in shipboard diarrheal disease outbreaks throughout Asia. A large outbreak of suspected Norwalk virus was investigated on a U.S. Naval aircraft carrier following the clinical recognition of 450 cases of gastroenteritis over a 2-week period (September 14-28, 1997) during coastal exercises. A random sampling of 44 cases from 450 personnel who sought medical attention was compared with 19 controls. Junior enlisted sailors and marines comprised 97% of all cases. There was no evidence of shipboard geographic clustering of cases. Furthermore, no single food type was associated with illness on the basis of comparative analysis (cases versus controls). Principal case signs and symptoms reported included watery stools (89%), nausea (82%), and vomiting (77%). Anecdotal reports indicated > 50% of the cases received rehydration therapy. An absence of fever was also noted in 32% of the cases and only 5% had blood in their stools. The mean duration of illness was 37 hr, with a range of 3-96 hr. Laboratory findings based on reverse transcription-polymerase chain reaction and Southern hybridization methods showed that 21 (72%) of 29 patients had evidence of the UK2 prototype of the Norwalk virus. A cross sectional study of 131 crew members from the ships population (n = 4,200) showed an attack rate of 44%. Attack rate is a variant of an incident rate applied to a narrowly defined population observed for a limited period of time, such as during an outbreak. The numerator is people who get sick and the denominator is people (population) at risk. An extrapolation of these findings suggests as many as 1,806 sailors may have been affected during the outbreak, of which only 26% (of the 57 outbreak related cases) where identified from sick call records. There was no difference in the mean ages between outbreak and non-outbreak affected crewmen, or geographic clustering based on berthing or work spaces. Outbreak related cases reported signs and symptoms of watery-stools (79%), nausea (65%), and vomiting (47%). The mean duration of illness was 28 hr, ranging from 2 to 96 hr. Thirty-one percent of outbreak affected cases reported a sick call visit. Loss of work was reported by 39% of the outbreak affected population. This report documents the epidemic potential of Norwalk virus and the associated impact on fleet operational readiness. Additionally, that this outbreak occurred against a background of 3 other consecutive gastroenteritis outbreaks onboard the same ship (March 1997, February/March 1998, and June 1998), all sharing the same clinical and epidemiologic profiles, suggests possible shipboard persistence of Norwalk virus over time, despite periodic ship-wide disinfection efforts. PMID- 10674668 TI - Norwalk-like virus and bacterial pathogens associated with cases of gastroenteritis onboard a US Navy ship. AB - Acute gastroenteritis is a potential cause of substantial morbidity in U.S. military personnel during deployment. This study investigated the microbial causes of diarrhea in U.S. troops on exercises in Southeast Asia aboard the U.S.S. Germantown from March through May 1996. A total of 49 (7%) patients with diarrhea reported to sick call during a 3-month deployment involving 721 personnel. Diarrheal samples from 49 patients were subjected to bacterial and parasitologic examination, but sufficient samples from only 47 of 49 were available for analysis of the presence of Norwalk-like virus (NLV). Of the 49 diarrhea cases, 10 (20.4%) appeared to be due to bacterial etiology alone, 10 (20.4%) due to bacteria and the prototype Taunton agent (TNA), 11 (22.4%) due to TNA only, and 4 (8.0%) due to parasites. Norwalk-like virus RNA was present in 21 (45%) of 47 stool samples from the diarrhea cases, 10 with bacterial etiologies and 11 without bacterial or parasitic etiologies. No pathogen was detected in 14 (29%) of the cases. Four of the controls showed the presence of parasitic organisms. Of the 11 cases in which enterotoxigenic Escherichia coli was isolated, 8 were positive for colonization factor antigen (CFA/IV), and 3 were CFA-negative. The bacterial pathogens tested were all susceptible to gentamicin, and furadantin, but were resistant to ceftriaxone and norfloxacin, including 75% of the Campylobacter spp. These data support the view that the major cause of diarrhea for troops deployed in this geographic area is most likely NLVs. PMID- 10674669 TI - Epidemiology of the incidence of oro-facial noma: a study of cases in Dakar, Senegal, 1981-1993. AB - Oro-facial noma is an oral gangrene occurring in early childhood in extremely poor areas. As many as 70-90% of those with noma die, and to date, there is no satisfactory treatment to fight this disease. Within the context of the World Health Organization international program against noma, a 13-year retrospective study based on clinical records was carried out in Dakar, Senegal in an attempt to understand the epidemiology of noma. Between 1981 and 1993, 199 cases of noma were identified, among them; 36.7% were acute cases and 63.3% showed sequelae. Chronic sequelae of noma were seen in patients 2-41 years of age, but the acute phase of noma was found only in young children (77.7% in those 1-4 years of age, maximum age = 9 years, mean age +/- SD age = 3.4 +/- 1.9 years). A total of 73.1% of the cases with acute disease were reported in the Dakar, Diourbel and Kaolack regions during the dry season (57.0% of the cases). The lesions of progressive noma were localized mainly on the upper lip (42.4%) and the cheek (31.1%). A total of 96.9% of the patients with acute diseases were had poor general health with serious associated diseases; only 20.0% had a good vital prognosis. The development of epidemiologic surveillance programs for noma should be a public health priority in Senegal. PMID- 10674670 TI - Environmental conditions favoring bat infection with Histoplasma capsulatum in Mexican shelters. AB - Histoplasma capsulatum was isolated from gut, lung, liver, and spleen of 17 of 208 captured bats belonging to 6 different genera and species. Three of the 17 infected bats were from the State of Guerrero and 14 were from the State of Morelos. All were adult bats: 6 males (1 Pteronotus parnellii, 2 Natalus stramineus, 2 Artibeus hirsutus, and 1 Leptonycteris nivalis) and 11 females (1 Myotis californicus, 1 Mormoops megalophylla, 8 A. hirsutus, and 1 L. nivalis). High rates of bat infection with H. capsulatum were found in the monitored sites of the State of Morelos. Histoplasma infection of N. stramineus, A. hirsutus, and L. nivalis should be considered as the first records in the world. The fungus isolated from infected bats was identified by its typical mycelial-phase morphology and by its yeast-phase conversion. Exoantigen production confirmed the fungal identification by the presence of specific precipitation lines in double immunodiffusion assays using human immune serum. Histopathologic studies showed intracellular yeast-like cells compatible with H. capsulatum yeast-phase in tissues of several bats, especially in pulmonary (intra-alveolar and septal) macrophages, with none or minimal tissue reaction. In contrast to past reports, present data support a high risk of bat infection with H. capsulatum in Mexican cave environments. PMID- 10674671 TI - Infectious and tropical diseases in Oman: a review. AB - Oman is generally hot and dry, but the Salalah region in southern Dhofar province is relatively cool and rainy during the summer monsoon, and has a distinctive pattern of infection. Important, notifiable infections in Oman include tuberculosis, brucellosis (endemic in Dhofar), acute gastroenteritis, and viral hepatitis: 4.9% of the adults are seropositive for hepatitis B surface antigen and approximately 1.2% for hepatitis C virus. Infection with human immunodeficiency virus is uncommon, and leprosy, rabies, and Crimean-Congo hemorrhagic fever are rare. Between 1990 and 1998, the incidence of malaria, (>70% due to Plasmodium falciparum) decreased from 32,700 to 882 cases. Cutaneous and visceral leishmaniasis (caused by Leishmania tropica and L. infantum, respectively) and Bancroftian filariasis occur sporadically. Intestinal parasitism ranges from 17% to 42% in different populations. A solitary focus of schistosomiasis mansoni in Dhofar has been eradicated. There are major programs for the elimination of tuberculosis, leprosy, and malaria, and to control brucellosis, leishmaniasis, sexually transmitted diseases, trachoma, acute respiratory infection in children, and diarrheal diseases. The Expanded Program on Immunization was introduced in 1981: diphtheria, neonatal tetanus, and probably poliomyelitis have been eliminated. PMID- 10674672 TI - Factors influencing resistance to reinfection with Plasmodium falciparum. AB - A treatment-reinfection study design was used to investigate the relationships between host immunologic and/or genetic factors and resistance to reinfection with Plasmodium falciparum. Sixty-one children in Gabon were enrolled in a cross sectional study to measure the prevalence of each human plasmodial species. All were given amodiaquine for radical cure of parasites, and 40 were subsequently followed-up for 30 weeks. Successive blood smears were examined to measure the delay of reappearance in blood of asexual stages of P. falciparum parasites. Presence of infection during the cross-sectional survey was associated with male sex, non-deficient glucose-6-phosphate dehydrogenase activity, plasma interleukin 10 level, and anti-LSA-Rep antibody concentration. Resistance to reinfection was related to the presence of anti-LSA-J antibodies, and the absence of anti-LSA-Rep antibodies. Moreover, P. malariae-infected subjects were usually co-infected with P. falciparum, and were also more rapidly reinfected with P. falciparum after treatment, compared with those without P. malariae infection. PMID- 10674673 TI - Analysis of repeated hemoglobin measures in full-term, normal birth weight Kenyan children between birth and four years of age. III. The Asemobo Bay Cohort Project. AB - Anemia is an important public health problem. During very early childhood numerous factors affect hemoglobin (Hb) concentration over time, making single cross-sectional measurements difficult to interpret when studying the natural history of anemia or evaluating anemia control strategies. We analyzed repeated Hb measures contributed by 942 Kenyan children between birth and 48 months of life using a mixed effects model, with a regression spline used to describe the population mean Hb profile, and random intercepts and slopes and first-order autoregressive correlation structure to accommodate the within-individual correlation among the repeated Hb measures. The approach facilitates the study of time-stationary and time-varying covariates that influence Hb in early life. The fitted mean Hb profile obtained from the analytic model is consistent with the observed mean Hb of the study population. Village of residence was associated with greatest difference in mean Hb at time of birth (16 versus 19 g/dL; P < 0.0001). Monthly weight-for-age was also associated with mean Hb after 3 months of age. This is the first description of an analysis strategy specifically for repeated Hb measures collected in a longitudinal field study in Africa. The strategy will facilitate improved study of time-varying covariates thought to influence pediatric anemia. PMID- 10674674 TI - Kala-azar in a high transmission focus: an ethnic and geographic dimension. AB - In 1994-1996, we studied a group of 58 game wardens stationed in an area known to be highly endemic for visceral leishmaniasis (kala-azar) for evidence of infection with Leishmania donovani. Leishmania DNA was detected by the polymerase chain reaction in the peripheral blood of cases of active kala-azar, former patients with visceral leishmaniasis, patients, and asymptomatic subjects. Using the cloned antigen rk39, antibodies were detected in 44.2% of the game wardens while leishmanin skin test result was positive in 77% of our sample. It was shown that certain tribes from northern Sudan were more likely to develop subclinical infections, while those of the Baria tribe from southern Sudan and those of the Nuba tribe from western Sudan were more likely to develop visceral leishmaniasis. Whether this is due to genetic factors or previous exposure to Leishmania parasites remains to be elucidated. PMID- 10674675 TI - Epidemiologic aspects of American visceral leishmaniasis in an endemic focus in Eastern Venezuela. AB - An endemic focus of American visceral leishmaniasis (AVL) in eastern Venezuela has been evaluated in terms of patients (n = 48), immunologic reactivity to Leishmania in household contacts (n = 187) and neighborhood controls (n = 170), detection of Leishmania (L. donovani complex) in dogs and wild animals by the polymerase chain reaction (PCR) and characteristics of the sandfly population. The male:female ratio of patients was 1.18:1; 89.6% were < or =12 years old. Serologic reactivity was significantly higher in household contacts than in controls (P = 0.0008), as was the size of leishmanin reactions in contacts < or =10 years of age (P = 0.0141). Leishmania donovani complex-specific PCRs were positive in dogs, an opossum (Didelphis marsupialis), and a black rat (Rattus rattus). Lutzomyia longipalpis and Lu. evansi, both implicated in the transmission of AVL, were identified among the 386 sand flies examined. These observations provide the bases for an active control program as well as further studies of reservoirs and vector-host relationships in this area. PMID- 10674676 TI - Geographic distribution and epidemiology of Oesophagostomum bifurcum and hookworm infections in humans in Togo. AB - In contrast to the rest of the world, infections with Oesophagostomum bifurcum are commonly found in humans in northern Togo and Ghana. In addition, infections with hookworm are endemic in this region. In the present study, a detailed map of the geographic distribution of O. bifurcum and hookworm infections in northern Togo was made. There were a number of foci with high prevalence of infection with O. bifurcum. All the villages examined were infected with hookworm, and the distribution was quite patchy. Women were infected with O. bifurcum more often than men, while infections with hookworm were more prevalent in men than in women. The prevalence and intensity of infection with both parasites were clearly age-dependent. We estimate that more than a 100,000 people in Togo are infected with O. bifurcum and more than 230,000 are infected with hookworm. PMID- 10674677 TI - Use of polymerase chain reaction for accurate follow-up of Loa loa experimental infection in Mandrillus sphinx. AB - Mandrills (Mandrillus sphinx) experimentally infected with human Loa loa usually remain microfilaremic for a long period of time. Nevertheless some control their microfilaremia while still harboring adults worms, and therefore become occult infected. A nested polymerase chain reaction (PCR) assay, targeted on the repeat 3 region of the gene coding for the L. loa 15-kD protein (15r3-PCR), has been evaluated in mandrills infected with third-stage larvae (L3) of L. loa. The results of this assay were negative during the prepatency period (4 months after inoculation), but became positive when microfilariae appeared in the blood, and remained positive in all mandrills, even in those that became amicrofilaremic. These results show that the positivity of the 15r3-PCR assay is linked to the appearance of microfilariae in peripheral blood and demonstrated that L. loa specific DNA can be detected in blood from occult-infected mandrills. PMID- 10674678 TI - Response of cutaneous leishmaniasis (chiclero's ulcer) to treatment with meglumine antimoniate in Southeast Mexico. AB - Cutaneous leishmaniasis, known as chiclero's ulcer in southeastern Mexico, is characterized by a predominantly single, painless, ulcerated lesion, without lymphangitis or adenopathy. When located on the ear, it tends to become chronic, causing destruction of the pinna and disfigurement. It is caused predominantly by Leishmania (L.) mexicana. Although pentavalent antimonials (Sb5+) are the mainstay of leishmanial therapy and have been used for more than 50 years, dosage regimens have been repeatedly modified and the best one has not been fully identified. The main purpose of the present study was to investigate the response of chiclero's ulcer to treatment with meglumine antimoniate. One hundred five patients were treated with meglumine antimoniate at a daily dose of 1 ampule per day (425 mg of Sb5+) until healing. The lesions healed after a mean of 25 days (range = 5-60 days). PMID- 10674679 TI - No evidence of cardiotoxicity during antimalarial treatment with artemether lumefantrine. AB - Artemether-lumefantrine is a new fixed antimalarial combination effective against multidrug-resistant falciparum malaria. A prospective electrocardiographic study was conducted in 150 patients receiving artemetherlumefantrine and 50 treated with artesunate-mefloquine. There was no evidence for clinically significant changes in the electrocardiographic intervals and in particular no relationship between plasma concentrations of lumefantrine and QTc prolongation. Artemether lumefantrine does not have significant cardiac effects at therapeutic doses. PMID- 10674680 TI - Assessment of therapeutic response of Plasmodium falciparum to chloroquine and sulfadoxine-pyrimethamine in an area of low malaria transmission in Colombia. AB - Although chloroquine (CQ) resistance was first reported in Colombia in 1961 and sulfadoxine-pyrimethamine (SP) resistance in 1981, the frequency of treatment failures to these drugs in Colombia is unclear. A modified World Health Organization 14-day in vivo drug efficacy test for uncomplicated Plasmodium falciparum malaria in areas with intense malaria transmission was adapted to reflect the clinical and epidemiologic features of a low-intensity malaria transmission area in the Pacific Coast Region of Colombia. Patients > or =1 year of age with a parasite density > or =1,000 asexual parasites per microliter were enrolled in this study. Forty-four percent (24 of 54) of the CQ-treated patients were therapeutic failures, including 7 early treatment failures (ETFs) and 17 late treatment failures (LTFs). Four (6%) of 67 SP-treated patients were therapeutic failures (2 ETFs and 2 LTFs). Therapeutic failure in the CQ-treated group was associated with an age <15 years old (P < 0.01), but was not associated with initial parasite density, the presence of CQ or sulfa-containing drugs in urine, or a history of malaria. The high level of therapeutic failures to CQ detected in this study underscores the need and importance of drug efficacy evaluation in the development of a rational national antimalarial drug policy. The relatively low level of therapeutic failures to SP compared with other South American countries raises further questions regarding factors that might have prevented the rapid development of in vivo resistance to this drug combination. PMID- 10674681 TI - Efficacy of primaquine regimens for primaquine-resistant Plasmodium vivax malaria in Thailand. AB - To define the current efficacy of Fansidar (F. Hoffmann-La Roche Ltd., Basel Switzerland) (pyrimethamine and sulfadoxine), primaquine in a high dose, and artesunate for treating acute Plasmodium vivax malaria, we conducted a comparative clinical trial of these 3 drugs in an open-label study. Patients (15 65 years old) were assigned to 1 of 4 treatments regimens in a serial order. Ninety percent of the patients were infected at Thailand-Myanmar border. Patients in group I (n = 23) received Fansidar (3 tablets, 75 mg of pyrimethamine and 1,500 mg of sulfadoxine, a single dose on the first day), group II (n = 23) received Fansidar (3 tablets, 75 mg of pyrimethamine and 1,500 mg of sulfadoxine, a single dose on the first day) and then received primaquine (30 mg a day for 14 days), group III (n = 23) received primaquine (30 mg a day for 14 days), and group IV (n = 23) received artesunate (200 mg once a day for 3 days) and then primaquine (30 mg a day for 14 days). Cure rates on day 28 of follow-up were 40%, 100%, 100%, and 100% in groups I, II, II, and IV, respectively. There were 4 and 5 patients in group I showing post-treatment reappearance of parasitemia at < or = 16 days and between 17 and 28 days, respectively. Patients in the other 3 groups showed negative parasitemias within 7 days after treatment. Artesunate plus primaquine (group IV) cleared parasitemia faster than the other 3 regimens. There is a high proportion of ineffectiveness of Fansidar for treatment of P. vivax malaria and it should be no longer used for treatment of P. vivax malaria acquired at the Thailand-Myanmar border. A high dose of primaquine is safe and effective in the treatment of P. vivax malaria during the 28-day follow-up period. PMID- 10674682 TI - Plasmodium falciparum parasites in French Guiana: limited genetic diversity and high selfing rate. AB - The genetic characteristics of Plasmodium falciparum isolates collected in French Guiana, where malaria transmission is low and occurs in isolated foci, were studied. Blood samples were collected from 142 patients with symptomatic malaria and typed using a polymerase chain reaction-based strategy for merozoite surface protein-(MSP-1) block 2, the MSP-2 central domain, and glutamate-rich protein (GLURP) repeat domain polymorphism. This showed that the parasite population circulating in French Guiana presented a limited number of allelic forms (4, 2, and 3 for MSP-1 block 2, MSP-1, and GLURP, respectively) and a small number of mixed infections, contrasting with the large genetic diversity of parasite populations and infection complexity reported for Africa, Asia, and other parts of South America. Two groups of isolates displaying identical 3 loci allele combinations were further studied for the Pf332 antigen, histidine-rich protein 1, thrombospondin-related anonymous protein, and Pf60 multigene family polymorphism. Within each group, most isolates were identical for all markers tested. This suggests a high rate of self-fertilization of P. falciparum parasites in French Guiana, resulting in homogenization of the population. The implications of these findings for malaria control in areas of low endemicity are discussed. PMID- 10674683 TI - Genetic structure and phylogenetic relationships of Colombian Trypanosoma cruzi populations as determined by schizodeme and isoenzyme markers. AB - Twenty-four Trypanosoma cruzi stocks isolated from vectors and from human and Didelphis marsupialis hosts from highly separated sylvatic localities in Colombia were characterized by isoenzyme and schizodeme analyses. The stocks were collected primarily from sylvatic ecotopes representing areas of low, moderate, and high endemicity for Chagas' diseases in Colombia. Parasites were characterized mainly by schizodeme analysis with the restriction enzyme Eco RI and the isoenzyme analysis was performed at 10 genetic loci. These analyses demonstrated an agreement between the classifications based on the isoenzyme analysis and on the restriction fragment length polymorphism patterns obtained with the Colombian stocks. There is clear evidence of demic subdivision between the eastern (E) and western (W) stocks separated by the Andean Mountains and Magdalena River, which is likely due to the geographic isolation generated by these topographic features. Heterozygosity estimates indicate that the E group could be more ancient than the W group. As was postulated in a previous study, these results are also compatible with the existence of a clonal population structure in Colombian sylvatic T. cruzi. Evidence presented here failed to demonstrate a correlation between the degree of endemy and genetic clustering. Finally, schizodeme and isoenzymatic analyses comparing Colombian T. cruzi stocks with others from Chile confirm that Colombian isolates are genetically related to zymodeme 1 and distant from zymodeme 2. PMID- 10674684 TI - Detection and genetic relationship of dengue virus sequences in seventeen-year old paraffin-embedded samples from Cuba. AB - This study describes the use of the reverse transcriptase-polymerase chain reaction (RT-PCR) to generate dengue 2 amplicons from paraffin-embedded autopsy tissues collected in Cuba 17 years ago. The presumptive diagnoses had been made only by clinical evolution without serologic confirmation. This study confirms once again that dengue 2 virus was directly associated with the fatal cases in children and illustrates the potential of the RT-PCR for retrospective diagnosis of dengue cases 17 years after death. A close similarity in the genomic sequences of the dengue 2 RNA detected in tissue samples from fatal cases and those dengue 2 Cuban strains that had been previously investigated confirms the appropriate genomic classification of the etiologic agent associated with the 1981 dengue hemorrhagic fever Cuban epidemic. PMID- 10674685 TI - Short report: microsatellite sequences as markers for population genetic studies of the mosquito Aedes aegypti, the vector of dengue viruses. AB - We report the isolation of microsatellites from an enriched library of genomic repeated sequences, using a biotin-labeled oligonucleotide bound to streptavidin coated magnetic particles. Four microsatellites were obtained from a partial library of 120 recombinant clones. This more efficient and rapid method to obtain these specific repeated sequences is preferred to the conventional isolation procedure based on the construction of a genomic library. Microsatellite markers would be promising molecular tools for the study of genetic variability of mosquito populations. Analyses of genetic structure and gene flow would provide information on the distance, direction and rate of dispersal of genes in Aedes aegypti populations. Knowledge on gene dispersal patterns is required to develop vector control strategies. PMID- 10674686 TI - Occurrence of sibling species of Lutzomyia longipalpis (Diptera: Psychodidae) in Venezuela: first evidence from reproductively isolated sympatric populations. AB - The delimitation of cryptic species within the main vector of the American visceral leishmaniasis, Lutzomyia longipalpis, remains a topic of controversy. An analysis of genetic variability based on 8 enzymatic loci revealed fixed differences in 2 diagnostic loci, adenylate kinase (Ak) and hexokinase (Hk), between sympatric and allopatric populations at 4 localities in Venezuela. The absence of heterozygotes for these 2 loci within 1 locality indicates, for the first time, the presence of 2 sympatric reproductively isolated populations or cryptic species within L. longipalpis. Significant differences were also detected between these cryptic species in the allele frequencies of glucose-6-phosphate isomerase (Gpi) and malate dehydrogenase, decarboxylating (Me). One species showed mean heterozygosities that ranged between 6.6% and 6.7%, with 1.6-1.9 alleles detected per locus, while the other had mean heterozygosities that ranged from 4.3% to 6.3%, with 1.3-1.6 alleles per locus. Comparisons of isozyme profiles with published data suggests that 1 species is similar to the L. longipalpis described in Colombian and Brazilian populations, whereas the other has not been previously reported. PMID- 10674687 TI - Spatial distribution and habitat characterization of anopheline mosquito larvae in Western Kenya. AB - Studies were conducted to characterize larval habitats of anopheline mosquitoes and to analyze spatial heterogeneity of mosquito species in the Suba District of western Kenya. A total of 128 aquatic habitats containing mosquito larvae were sampled, and 2,209 anopheline and 10,538 culicine larvae were collected. The habitats were characterized based on size, pH, distance to the nearest house and to the shore of Lake Victoria, coverage of canopy, surface debris, algae and emergent plants, turbidity, substrate, and habitat types. Microscopic identification of third- and fourth-instar anopheline larvae did not yield any Anopheles funestus or other anophelines. A total of 829 An. gambiae s.l. larvae from all habitats were analyzed further by rDNA-polymerase chain reaction to identify individual species within the An. gambiae species complex. Overall, An. arabiensis was the predominant species (63.4%), and An. gambiae was less common (31.4%). The species composition of An. gambiae s.l. varied significantly among the sampling sites throughout Suba District. The larval habitats in the southern area of the district had a higher proportion of An. gambiae than in the northern area. Multiple logistic analysis did not detect any significant association between the occurrence of anopheline larvae and habitat variables, and principal component analysis did not identify key environmental factors associated with the abundance of An. gambiae. However, significant spatial heterogeneity in the relative abundance of An. gambiae within the Suba district was detected. When the effect of larval habitat locality was considered in the analysis, we found that the distance to the nearest house and substrate type were significantly associated with the relative abundance of An. gambiae. Future studies integrating detailed water chemistry analysis, remote sensing technology, and the ecology of predators may be required to further elucidate the mechanisms underlying the observed spatial variation of anopheline larval distribution. PMID- 10674688 TI - Short report: treatment of snake envenomations by a new polyvalent antivenom composed of highly purified F(ab)2: results of a clinical trial in northern Cameroon. AB - A clinical trial was conducted in 2 health centers in northern Cameroon to assess the safety and efficacy of a new polyvalent antivenom composed of highly purified and pasteurized F(ab')2 (FAV-Africa). Forty-six patients with objective signs of envenomation, including 67% with hemorrhage, were included in the study. Each patient received at least 20 ml of FAV-Africa by direct, slow intravenous injection; 172 10-ml ampules were administered. All patients were clinically cured after treatment. Two patients (4.3%) showed minor immediate adverse events that may have been related to FAV-Africa (induration, light-headedness); no other treatment-related adverse event occurred. No patient had serum sickness. This trial confirms the safety of FAV-Africa administered by intravenous injection and its efficacy in the treatment of snake envenomations in sub-Saharan Africa. PMID- 10674689 TI - The International Health Program: the fifteen-year experience with Yale University's Internal Medicine Residency Program. AB - The purpose of this study was to assess the impact of international health electives on physicians-in-training. A retrospective study was conducted using an anonymous, self-administered mailed survey to internal medicine residents who trained at Yale from 1982 to 1996 based on their experience with our International Health Program (IHP). The response rate was 61%, with 96 completed surveys in the participant group and 96 completed surveys in the nonparticipant group. Participants were more likely than nonparticipants to care for patients on public assistance (77 versus 49; P < 0.001) and immigrant patients (41 versus 23; P = 0.006). Among residents who changed their career plans, participants (22) were more likely than nonparticipants (14) to switch from subspecialty medicine to general medicine (P = 0.02). Participants were significantly more likely to have a positive view of health care delivery in developing countries. Compared with nonparticipants (64), IHP participants (74) believed that the physical examination is under-used by physicians from the United States as a diagnostic skill (P = 0.03). International health experiences appeared to have an important impact on the decisions and attitudes of residents. PMID- 10674690 TI - Call for 'radical overhaul' of meat hygiene arrangements. PMID- 10674691 TI - Therapeutic efficacy of water-soluble lincomycin-spectinomycin powder against porcine proliferation enteropathy in a European field study. AB - Controlled clinical trials to a standardised protocol were conducted into the effect of a water-soluble antibiotic on proliferative enteropathy and its causative agent (Lawsonia intracellularis) on commercial pig farms at six sites in four European countries. Clinical signs of the disease and L intracellularis specific polymerase chain reaction (PCR)-positive pigs were detected in pens of six- to 12-week-old pigs (weighing 5 to 55 kg) immediately before each trial. Matched pens of randomised pigs were either left unmedicated (32 to 59 pigs per trial), or medicated orally with 10 mg/kg of a water-soluble combination of lincomycin and spectinomycin powder (21 and 42 mg, respectively, of antibiotic activity per litre) for either seven days (33 to 61 pigs per trial), or 14 days (33 to 61 pigs per trial), delivered via the drinking water. Investigators did not know which pens received which treatment In most of the affected pigs in each trial, diarrhoea due to L intracellularis resolved within three to seven days after the medication began, whereas most unmedicated pigs remained diarrhoeic for at least 10 days. On average the medicated pigs gained more weight than the unmedicated pigs over the 21-day trial period (P=0.01). In two trials, the absence of L intracellularis after the treatment ended was confirmed by the PCR. PMID- 10674692 TI - Survey of the effectiveness of stunning procedures used in Spanish pig abattoirs. AB - Two pig abattoirs (A and B) equipped with an automated head-only and head-to chest electrical stunning system, and two (C and D) equipped with a manual carbon dioxide stunning system, were evaluated to compare the effectiveness of stunning in a total of 10,454 pigs slaughtered under commercial conditions. In the abattoirs with the electrical stunning system, the percentage of animals that responded to a nose prick was significantly lower (P<0.05) in abattoir B, where a higher current intensity was used (P<0.05), than in abattoir A. No righting reflex was observed in the electrically stunned pigs. In the abattoirs with the carbon dioxide stunning system, the percentage of animals that responded to a nose prick and showed a righting reflex was significantly lower (P<0.05) in abattoir C, where the duration of the carbon dioxide cycle was longer and the interval between discharge from the system to sticking was shorter (P<0.05), than in D. Comparing the electrical and carbon dioxide stunning systems, the pigs stunned with carbon dioxide were significantly more responsive to a nose prick (P<0.05) and 25 per cent of them showed a righting reflex. Under the conditions of the study the fully automated head-only stunning with additional chest electrodes appeared to be more effective and less susceptible to incorrect handling than the manual carbon dioxide stunning system. PMID- 10674694 TI - Phytophotodermatitis in pigs exposed to parsley (Petroselinum crispum). PMID- 10674693 TI - Evidence of Muscovy duck parvovirus in Muscovy ducklings in California. AB - Muscovy duck parvovirus (MDPV) has been demonstrated in tissue samples from one- to four-week-old commercially reared Muscovy ducks that were weak, unable to walk and had a high mortality rate. On postmortem examination, the thigh and leg muscles, and the myocardium were found to be pale, and there was a fibrinous exudate on the capsule of the liver, and ascites. The parvovirus was isolated in embryonated Muscovy duck eggs and visualised by negative stain electron microscopy, detected by polymerase chain reaction (PCR) directly from the tissues, and antibodies to it were detected by immunoelectron microscopy, ELISA and immunofluorescence. In addition, the PCR products obtained that represented 1625 bp (74 per cent) of the capsid vP1 gene, including a hypervariable region between Derzsy's disease virus or goose parvovirus and MDPV, were sequenced and shown to be 100 per cent homologous with the MDPV 89384 reference strain, but only 82.3 per cent homologous with Derzsy's disease virus. PMID- 10674695 TI - Efficiency of Western blotting for the specific immunodetection of proteinase K resistant prion protein in BSE diagnosis in France. PMID- 10674696 TI - Emergence of resistance to fluoroquinolones among bacteria causing infections in food animals in Denmark. PMID- 10674697 TI - Hemiovariosalpingectomy in a loggerhead sea turtle (Caretta caretta). PMID- 10674698 TI - Health status of dairy herds in organic farming. PMID- 10674699 TI - Corynebacterium renale infection in calves. PMID- 10674700 TI - Postweaning multisystemic wasting syndrome of pigs. PMID- 10674701 TI - Society of Veterinary Assistants. PMID- 10674703 TI - Reindeer concerns PMID- 10674702 TI - Reindeer concerns. PMID- 10674704 TI - Reindeer concerns PMID- 10674705 TI - Intravitreal administration of antisense oligonucleotides: potential of liposomal delivery. AB - Antisense oligonucleotides are short synthetic fragments of genes that are able to inhibit gene expression after being internalized by cells. They can therefore be used as antiviral compounds particularly, for the treatment of ocular viral infections (i.e. Herpes simplex virus or Cytomegalovirus, CMV). Antisense oligonucleotides are however poorly stable in biological fluids and their intracellular penetration is limited. Although oligonucleotides are now currently used in therapeutics for the treatment of CMV by intravitreal injection (Vitravene) their main drawbacks impose to repeat the number of administrations which can be very harmful and damaging. A system that is able to permit a protection of oligonucleotides against degradation and their slow delivery into the vitreous would be more favorable for improving patient compliance. The use of liposomes for intravitreal administration can be very promising since these lipid vesicles are able to protect oligonucleotides against degradation by nucleases and they allow to increase the retention time of many drugs in the vitreous. In this review, the potentialities of liposomes for the intravitreal delivery of oligonucleotides will be discussed. PMID- 10674706 TI - Development and clinical assessment of an artificial cornea. AB - Keratoprosthesis research has been a gradual, rather fragmentary process with advances being made by isolated groups of researchers. This has arisen partly because of poor funding in the area; research groups which have achieved commercial support have often had constraints upon the full disclosure of their findings. Despite these difficulties there has been real progress over the last decade by several independent groups. This article concentrates upon our own development of a hydrogel core-and-skirt keratoprosthesis, the Chirila KPro, in order to illustrate the scientific and clinical problems common to keratoprosthesis research. Pilot data from a clinical trial is presented and the priorities for future research are discussed. PMID- 10674707 TI - Role of Schwann cells in retinal ganglion cell axon regeneration. AB - It is a well known fact that the injured PNS can successfully regenerate, on the other hand, the CNS such as retinal ganglion cell (RGC) axons of adult mammals is incapable of regeneration. After injury, RGC axons rapidly degenerate and most cell bodies go through the process of cell death, while glial cells at the site of injury undergo a series of responses which underlie the so-called glial scar formation. However, it has become apparent that RGCs do have an intrinsic capacity to regenerate which can be elicited by experimental replacement of the inhibitory glial environment with a permissive peripheral nerve milieu. Schwann cells are a major component of the PNS and play a role in regeneration, by producing various kinds of functional substances such as diffusible neurotrophic factors, extracellular matrix and cell adhesion molecules. RGC regeneration can be induced by cooperation of these substances. The contact of RGC axons to Schwann cells based upon the structural and molecular linkages seems to be indispensable for the stable and successful regeneration. In addition to cell adhesion molecules such as NCAM and L1, data from our laboratory show that Schwann cells utilize short focal tight junctions to provide morphological stabilization of the contact with the elongating axon, as well as a small scale of gap junctions to facilitate traffic of substances between them. Moreover, our results show that modifications of functional properties in neighboring glial cells of optic nerve are induced by transplantation of Schwann cells. Astrocytes usually considered to form a glial scar guide the regenerating axons in cooperation with Schwann cells. A decrease of the oligodendrocyte marker O4 and migration of ED-1 positive macrophages is observed within the optic nerve stump. Accordingly, RGC regeneration is not a simple phenomenon of axonal elongation on the Schwann cell membrane, but is based on direct and dynamic communication between the axon and the Schwann cell, and is also accompanied by changes and responses among the glial cell populations, which may be partly associated with the mechanisms of optic nerve regeneration. PMID- 10674708 TI - Oxidative damage and protection of the RPE. AB - This review provides a model for the role of oxidative stress in the etiology of age-related macular degeneration (AMD). Epidemiological studies of diet, environmental and behavioral risk factors suggest that oxidative stress is a contributing factor of AMD. Pathological studies indicate that damage to the retinal pigment epithelium (RPE) is an early event in AMD. In vitro studies show that oxidant treated RPE cells undergo apoptosis, a possible mechanism by which RPE cells are lost during early phase of AMD. The main target of oxidative injury seems to be mitochondria, an organelle known to accumulate genomic damages in other postmitotic tissues during aging. The thiol antioxidant GSH and its amino acid precursors protect RPE cells from oxidant-induced apoptosis. Similar protection occurs with dietary enzyme inducers which increase GSH synthesis. These results indicate that therapeutic or nutritional intervention to enhance the GSH antioxidant capacity of RPE may provide an effective way to prevent or treat AMD. PMID- 10674709 TI - Stem cells and differentiation stages in the limbo-corneal epithelium. PMID- 10674710 TI - Molecular genetics of hepatic methionine adenosyltransferase deficiency. AB - Hepatic methionine adenosyltransferase (MAT) deficiency is caused by mutations in the human MAT1A gene that abolish or reduce hepatic MAT activity that catalyzes the synthesis of S-adenosylmethionine from methionine and ATP. This genetic disorder is characterized by isolated persistent hypermethioninemia in the absence of cystathionine beta-synthase deficiency, tyrosinemia, or liver disease. Depending on the nature of the genetic defect, hepatic MAT deficiency can be transmitted either as an autosomal recessive or dominant trait. Genetic analyses have revealed that mutations identified in the MAT1A gene only partially inactivate enzymatic activity, which is consistent with the fact that most hepatic MAT-deficient individuals are clinically well. Two hypermethioninemic individuals with null MAT1A mutations have developed neurological problems, including brain demyelination, although this correlation is by no means absolute. Presently, it is recommended that a DNA-based diagnosis should be performed for isolated hypermethioninemic individuals with unusually high plasma methionine levels to assess if therapy aimed at the prevention of neurological manifestations is warranted. PMID- 10674711 TI - Pharmacokinetics of selective serotonin reuptake inhibitors. AB - The five selective serotonin reuptake inhibitors (SSRIs), fluoxetine, fluvoxamine, paroxetine, sertraline, and citalopram, have similar antidepressant efficacy and a similar side effect profile. They differ, however, in their pharmacokinetic properties. Under steady-state concentrations, their half-lives range between 1 and 4 days for fluoxetine (7 and 15 days for norfluoxetine) and between 21 (paroxetine) and 36 (citalopram) hr for the other SSRIs. Sertraline and citalopram show linear and fluoxetine, fluvoxamine, and paroxetine nonlinear pharmacokinetics. SSRIs underlie an extensive metabolism with high interindividual variability, whereby cytochrome P450 (CYP) isoenzymes play a major role. Therefore, resulting blood concentrations are highly variable between individuals. Except for N-demethylated fluoxetine, metabolites of SSRIs do not contribute to clinical actions. Therapeutically effective blood concentrations are unclear so far, although there is evidence for minimal effective and upper threshold concentrations that should not be exceeded. Paroxetine and, to a lesser degree, fluoxetine and norfluoxetine are potent inhibitors of CYP2D6 and fluvoxamine of CYP1A2 and CYP2C19. This can give rise to drug-drug interactions that may have no effect, lead to intoxication, or improve the therapeutic response. These different pharmacokinetic properties of the five SSRIs, especially their drug-drug interaction potential, should be considered when selecting a distinct SSRI for treatment of depression or other disorders with a suggested dysfunction of the serotonergic system in the brain. PMID- 10674712 TI - Cardiotrophin-1: a novel cytokine and its effects in the heart and other tissues. AB - Cardiotrophin-1 (CT-1) originally was discovered as a factor that can induce hypertrophy of cardiac myocytes, both in vitro and in vivo. Subsequently, CT-1 has been shown to have a wide variety of different effects on cardiac and noncardiac, cells including the ability to stimulate the survival of both cardiac and neuronal cells. Like other members of the interleukin-6 family of cytokines, CT-1 stimulates both the p42/p44 mitogen-activated protein kinase pathway and the Janus-activated kinase/signal transducers and activators of transcription pathway. Interestingly, whilst activation of the p42/p44 mitogen-activated protein kinase pathway is necessary for the survival-promoting effects of CT-1 in cardiac cells, it is not required for its hypertrophic effect, which is likely to involve activation of the Janus-activated kinase/signal transducer and activator of transcription-3 pathway. CT-1, therefore, may be of use as a novel cardioprotective agent, particularly if its hypertrophic effect can be specifically inhibited. PMID- 10674713 TI - Molecular aspects of ATP-sensitive K+ channels in the cardiovascular system and K+ channel openers. AB - ATP-sensitive K+ (K(ATP)) channels are inhibited by intracellular ATP (ATPi) and activated by intracellular nucleoside diphosphates and thus, provide a link between cellular metabolism and excitability. K(ATP) channels are widely distributed in various tissues and may be associated with diverse cellular functions. In the heart, the K(ATP) channel appears to be activated during ischemic or hypoxic conditions, and may be responsible for the increase of K+ efflux and shortening of the action potential duration. Therefore, opening of this channel may result in cardioprotective, as well as proarrhythmic, effects. These channels are clearly heterogeneous. The cardiac K(ATP) channel is the prototype of K(ATP) channels possessing approximately 80 pS of single-channel conductance in the presence of approximately 150 mM extracellular K+ and opens spontaneously in the absence of ATPi. A vascular K(ATP) channel called a nucleoside diphosphate-dependent K+ (K(NDP)) channel exhibits properties significantly different from those of the cardiac K(ATP) channel. The K(NDP) channel has the single-channel conductance of approximately 30-40 pS in the presence of approximately 150 mM extracellular K+, is closed in the absence of ATPi, and requires intracellular nucleoside di- or triphosphates, including ATPi to open. Nevertheless, K(ATP) and K(NDP) channels are both activated by K+ channel openers, including pinacidil and nicorandil, and inhibited by sulfonylurea derivatives such as glibenclamide. It recently was found that the cardiac K(ATP) channel is composed of a sulfonylurea receptor (SUR)2A and a two transmembrane-type K+ channel subunit Kir6.2, while the vascular K(NDP) channel may be the complex of SUR2B and Kir6.1. By precisely comparing the functional properties of the SUR2A/Kir6.2 and the SUR2B/Kir6.1 channels, we shall show that the single-channel characteristics and pharmacological properties of SUR/Kir6.0 channels are determined by Kir and SUR subunits, respectively, while responses to intracellular nucleotides are determined by both SUR and Kir subunits. PMID- 10674714 TI - My journey from wool research to insulin. AB - This paper is an autobiographical study of the author's early work on the chemical cross-linking of proteins as well as on oligomer and peptide synthesis from 1949 in Heidelberg until the synthesis of insulin in 1963 in Aachen. PMID- 10674715 TI - Solid phase synthesis of peptide aldehyde protease inhibitors. Probing the proteolytic sites of hepatitis C virus polyprotein. AB - The solid phase synthesis of a set of peptide aldehydes derived from the NS5A/NS5B junction of hepatitis C virus (HCV) viral polyprotein is demonstrated using an oxazolidine linker and the Multipin method. Deletion of the P6 and P5 residues results in a dramatic loss of inhibitory activity. PMID- 10674716 TI - Chemical synthesis, characterization and activity of RK-1, a novel alpha-defensin related peptide. AB - The 32-residue peptide, RK-1, a novel kidney-derived three disulfide-bonded member of the antimicrobial alpha-defensin family, was synthesized by the continuous flow Fmoc-solid phase method. The crude, cleaved and S-reduced linear peptide was both efficiently folded and oxidized in an acidic solution of aqueous dimethyl sulfoxide. Following purification of the resulting product, it was shown by a variety of analytical techniques, including matrix assisted laser desorption time of flight mass spectrometry, to possess a very high degree of purity. The disulfide bond pairing of the synthetic peptide was determined by 1H-NMR spectroscopy and confirmed to be a Cys1-Cys6, Cys2-Cys4, Cys3-Cys5 arrangement similar to other mammalian alpha-defensin peptides. The synthetic RK-1 was also shown to inhibit the growth of Escherichia coli type strain NCTC 10418. PMID- 10674717 TI - Synthesis, conformational analysis and bioactivity of Lan-7, a lanthionine analog of TT-232. AB - A sandostatin analog, TT-232 (D-Phe-c[Cys-Tyr-D-Trp-Lys-Cys]-Thr-NH2), exhibits strong antitumor effects both in vitro and in vivo. In order to study the structure-activity relationships of TT-232, we designed and synthesized an analog of TT-232, namely Lan-7, in which the disulfide bridge is replaced by a lanthionine monosulfide bridge. Conformational analysis by NMR spectroscopy and computer simulations revealed that Lan-7 and TT-232 adopt very similar conformations in solution, which are quite different from the preferred conformations of sandostatin. Lan-7 has significant growth inhibition effects on a number of human tumor cell lines. It can also induce apoptosis in human ovarian carcinoma 2008 cells. At the same time, Lan-7 produced no toxicity to normal human hematopoietic progenitor cells. All of these results indicate that the modification we made does not alter the anti-tumor activity of TT-232. PMID- 10674718 TI - Synthesis of bivalent inhibitors of eucaryotic proteasomes. AB - Based on the peculiar spatial array of the active sites in the internal chamber of the multicatalytic proteasome, as derived from the X-ray structure of yeast proteasome, homo- and heterobivalent inhibitors were designed and synthesized to exploit the principle of multivalency for enhancing inhibition potency. Peptidic bis-aldehyde compounds of the octapeptide size were synthesized to address adjacent active sites, whilst a PEG spacer with a statistical length distribution of 19-25 monomers was used to link two identical or different tripeptide aldehydes as binding heads. These bis-aldehyde compounds were synthesized applying both methods in solution and solid phase peptide synthesis. Bivalent binding was observed only for the PEG-spaced inhibitors suggesting that binding from the primed side prevents hemiacetal formation with the active site threonine residue. PMID- 10674719 TI - Conization of the cervix using harmonic scalpel. AB - Conization, as a surgical treatment for cervical intraepithelial neoplasm (CIN), is a good method that preserves reproductive functions. Technological developments have introduced a wide variety of energy sources for surgical procedures. Traditional cold knife conization has been replaced by laser conization and by the loop electrosurgical excisional procedure (LEEP). However, laser conization and LEEP have some disadvantages. Laser conization requires expensive equipment. LEEP induces electrocautery artifacts and cannot excise the cervical tissue as a single-piece, because of the various extensions and depths of lesion, so that evaluation of the margins is sometimes not possible. Laser conization and LEEP both generate smoke. The presence of smoke is not only inconvenient, but also dangerous. Harmonic Scalpel (HS), ultrasonic cutting and coagulation system, is a new surgical tool that cuts and coagulates using ultrasonic mechanical vibrations. Eleven women with CIN III underwent conization using HS. HS eliminated the major disadvantages of electrosurgery and laser surgery. No complications during conization were observed. Postoperative hemorrhage was noted in only one patient. Histological diagnosis was not affected by heat or ultrasound. This surgical method using HS is characterized by negligible bleeding, a good visual field not obscured by smoke and resection of an ideal shape that fits the size of the lesion. It is concluded that this method overcomes most problems associated with conization using conventional methods. PMID- 10674720 TI - Cytokine gene expression after subretinal transplantation. AB - Transplantation study of neural retina, retinal pigment epithelial (RPE), or iris pigment epithelial (IPE) cells have been performed not only in animal model but in human age-related macular degeneration, and some of the findings reported with cystoid macular edema may have been due to graft rejection. In this investigation, we examined cytokine gene expression by reverse transcriptase polymerase chain reaction at the transplanted subretinal space. Transplantation was performed in normal Royal College of Surgeon's rats using cultured human RPE and rat IPE. They were followed without immunosupression. Gene expression for melanogenesis of transplanted human RPE was observed only in the early days after transplantation. Rat interleukin (IL)-1alpha, -1beta1, -2, -6, interferon gamma, and tumor necrosis factor alpha (TNF alpha) genes were also expressed after the early days of transplantation. Cytokine expression was observed not only after cell transplantation but also after vehicle-only injection, which was considered a reaction to the surgical trauma. However, statistically significant amount of expressions of IL-1alpha, -1beta, and -6 were observed after the early days of transplantation of human RPE or IL-1alpha, -1beta, and TNF alpha of rat IPE, if we compare them to vehicle-only injection. These cytokines may play an important role for the local reaction after transplantation. PMID- 10674721 TI - Interleukin-13 prevents diaphragm muscle deterioration in a septic animal model. AB - The effects of an intravenous injection of Interleukin-13 (IL-13) after endotoxin administration on diaphragm muscle were studied using Wistar rats. Two treatment groups, a control (saline+endotoxin) group and an IL-13 (IL-13+endotoxin) group were studied. E. coli endotoxin (10 mg/kg) was injected intraperitoneally 5 minutes after saline or IL-13 (0.25 microg) injection. The force-frequency curves, twitch kinetics and fatigability were measured at 0 and 4 hours after endotoxin injection. The force-frequency curves and twitch tension in the control group were significantly lower at 4 hours than those at 0 hour due to endotoxin. On the other hand, IL-13 prevented the decrement of the force-frequency curves and twitch tension induced by endotoxin. Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry showed positive staining at 4 hours due to endotoxin in the control group; however, IL-13 also blocked NADPH diaphorase staining at 4 hours. Furthermore, the positive muscle fibers detected by the NADPH diaphorase staining were classified as type I (slow twitch) muscle fibers by ATPase staining. We conclude that IL-13 prevents the deterioration of contraction induced by endotoxin by inhibiting nitric oxide production in the diaphragm muscle, mainly the type I muscle fibers. PMID- 10674722 TI - Therapeutic efficacy of transcranial magnetic stimulation for hereditary spinocerebellar degeneration. AB - We applied transcranial magnetic stimulation (TMS) as a therapeutic approach for patients with spinocerebellar degeneration (SCD). The subjects were four familial SCD patients (three men and one woman) aged from 27 to 76 years old. They were genetically analysed as two spinocerebellar ataxia type 6 (SCA 6), one SCA 1, and one SCA 7. The durations of their illness ranged from 1 to 7 years. Ten consecutive magnetic pulses were delivered over the scalp corresponding to the right cerebellar hemisphere, the middle of the cerebellum and the left cerebellar hemisphere, respectively, every day for 21 days. In all patients, the time and the number of steps required for a 10 m walk examination were significantly decreased after TMS trial compared with those before TMS. The number of feasible steps in tandem gait test increased. The total length of tracing body balance for 30 seconds measured by gravinometer was significantly decreased. However, nystagmus, dysarthria or incoordination of the upper limbs did not change after TMS trial. It is of interest that the blood flow of the cerebellar hemisphere, putamen and pons were significantly increased during the TMS trial. Although we do not know the exact mechanism by which TMS improved the ataxic gait, we speculate the increase of blood flow in the cerebellum, putamen and pons takes part in the improvement. These findings suggest that TMS over the cerebellum may be an effective therapy for patients with SCD. PMID- 10674723 TI - Stimulated neutrophils evoke signal transduction to increase vascular permeability in rat lungs. AB - The mechanisms by which stimulated neutrophils (PMNs) damage pulmonary vascular endothelium were investigated using twenty-four perfused lung preparations isolated from rats. We tested the ability of unstimulated and mechanically stimulated PMNs to adhere to pulmonary endothelial cells and, thereby, alter pulmonary vascular permeability (measured as the pulmonary filtration coefficient) and hemodynamics. To stimulate PMNs, they were gently agitated in a glass vial for 10 seconds. Perfusing lungs with the stimulated PMNs (stimulated group) elicited a 3-fold increase in the filtration coefficient as compared to lungs perfused with unstimulated cells (unstimulated group). This increase in filtration was completely blocked by preincubation of stimulated PMNs with CD18 monoclonal antibody (MoAb group). This increase in filtration coefficient was also completely blocked by GF109203X, a protein kinase C inhibitor (GF group). Pulmonary vascular resistance increased when the stimulated PMNs were injected to the isolated lungs. Although, preincubation of stimulated PMNs with CD18 MoAb successfully blocked and GF109203X partly blocked this increase in pulmonary vascular resistance. The accumulation of stimulated PMNs within the lungs, as assessed by myeloperoxidase (MPO) levels, was blocked by preincubation of stimulated PMNs with CD18 MoAb. However, GF109203X did not decrease MPO levels. These findings suggest that stimulated PMN-induced increases in pulmonary vascular filtration, resulted from endothelial cell injury caused by adhesion to the endothelial cells, evoke intracellular signaling within the endothelial cells. PMID- 10674724 TI - Fibrous dysplasia arising from the calcaneus. AB - A case of an 18-year-old woman with fibrous dysplasia arising in the calcaneus, which is extremely rare, is reported, with the emphasis placed on differential diagnosis from low-grade central osteosarcoma. She had a severe pain in her left ankle after sprain. Plain radiographs showed a radiolucent lesion measuring 6.3 x 2.5 cm with a sclerotic margin in the left calcaneus. CT scans showed a well defined lytic lesion with disruption of the lateral cortex and an ossification or calcification in its center. On MR imaging, the lesion had isointensities and high intensities on T1 and T2 weighted images, respectively, but its central portions showed lower intensities both on T1 and T2 weighted images. The lesion was enhanced with gadolinium except for the central portions. The specimen obtained by open biopsy consisted of fibrous tissue and foci of irregular woven bone. None of the nuclear atypia, mitoses, longitudinal stream of bone or invasive nature of growth was detected. The diagnosis of fibrous dysplasia was histologically made. The lesion was curetted and packed with autogenous bone chips. No evidence of recurrence was noted postoperatively. PMID- 10674725 TI - The existential ophthalmologist. PMID- 10674726 TI - Clinical research in oculoplastic surgery for the 21st century. PMID- 10674727 TI - The anatomy of midfacial ptosis. AB - PURPOSE: To investigate the anatomic and histologic changes present in midfacial ptosis. METHODS: Experimental study applying gross anatomic and histologic techniques to formalin-preserved and fresh-frozen cadaver heads with and without midfacial ptosis. High-resolution surface coil magnetic resonance imaging (MRI) was performed to obtain radiologic correlations. RESULTS: The orbitomalar ligament was further characterized by identification of a well-developed lateral component in the sub-superficial musculoaponeurotic plane; abnormalities of this important supporting structure were present in the subcutaneous plane in 8 of 10 specimens with midfacial ptosis. The zygomatic and masseteric cutaneous ligaments also were further characterized on a gross anatomic level, and histologic evidence of these two structures was produced. The subcutaneous components of the zygomatic and masseteric cutaneous ligaments were attenuated or not identifiable in 40% and 30% of specimens with midfacial ptosis, respectively. High-resolution surface coil MRI provided exquisite correlations of midfacial anatomy. CONCLUSIONS: The lateral component of the orbitomalar ligament provides major osteocutaneous midfacial support. Subcutaneous attenuation of the orbitomalar, masseteric cutaneous, and zygomatic ligaments was associated with midfacial ptosis. PMID- 10674728 TI - Injectable lyophilized particulate human fascia lata (Fascian) for lip, perioral, and glabellar enhancement. PMID- 10674729 TI - Small incision nonendoscopic browlift. AB - PURPOSE: To determine the efficacy of a nonendoscopic brow/forehead lift. METHODS: Case series of 12 patients. Small scalp and upper eyelid blepharoplasty incisions were used to elevate the brows/forehead and perform protractor myectomy. RESULTS: All patients achieved an aesthetically pleasing eyebrow and forehead lift with reduction of vertical and horizontal glabellar creases. Complications included one patient who experienced prolonged ecchymosis after corrugator muscle resection and one patient who had asymmetric lid folds after surgery. The length of follow-up ranged from 9 to 35 months. CONCLUSIONS: The small incision nonendoscopic browlift technique provides a useful alternative to the endoscopic approach. PMID- 10674730 TI - Frontalis suspension combined with blepharoplasty as an effective treatment for blepharospasm associated with apraxia of eyelid opening. AB - PURPOSE: Essential blepharospasm can be associated with apraxia of eyelid opening and is characterized by the inability to initiate the act of eyelid elevation even after cessation of orbicularis spasms. Current therapies such as botulinum toxin injections, orbicularis resection, or neurectomy may be unsuccessful or have undesired side effects. METHODS: Frontalis suspension was used to treat 13 consecutive patients with apraxia and blepharospasm during a 4-year interval. Follow-up ranged from 16 months to 55 months. To improve the aesthetic outcome, an upper blepharoplasty was done at the same time as the frontalis suspension in 7 cases. RESULTS: Good or excellent functional results were obtained in 10 of 13 patients. In 6 of these patients, the spasm disappeared completely. Therapy was unsuccessful in 1 patient, and in 2 patients blepharospasm recurred after 9 months. CONCLUSION: Patients with blepharospasm and apraxia of eyelid opening may benefit from a frontalis suspension operation, which can be considered minimally invasive and reversible. PMID- 10674731 TI - The tarsal sandwich: a new technique in lateral canthoplasty. AB - PURPOSE: Trauma and infection sometimes produce lower lid malpositions that are difficult to repair cosmetically with standard canthoplasty techniques. A new variation is described. METHODS: Surgical techniques of the tarsal strip canthoplasty and of lateral tarsorrhaphy are combined into the tarsal sandwich. RESULTS: Representative cases with preoperative and postoperative photos are presented. CONCLUSION: The sandwich technique allows the surgeon more flexibility in achieving the necessary vertical lift of the lateral canthus in difficult cases of entropion, ectropion, and lagophthalmos. PMID- 10674732 TI - Upper eyelid island orbicularis oculi myocutaneous flap for periorbital reconstruction. AB - PURPOSE: To describe the upper eyelid island orbicularis oculi myocutaneous flap, medially or laterally based, for reconstruction of periorbital defects. METHODS: During the past three years we have used the island orbicularis oculi myocutaneous flap in 62 patients with tumors of the periorbital area, with the following indications: (a) anterior lamellar defects of the medial aspect of the upper eyelid, when the peripheral arcade is intact; (b) up to 2/3 anterior lamellar lower eyelid defects; (c) inner and outer canthus defects; and (d) defects of the peripalpebral area (the lateral half of the eyebrow, bridge of the nose and suprazygomatic areas). RESULTS: The flap proved to be flexible, safe, relatively simple, and provided good functional and aesthetic results. Complications were minimal. CONCLUSIONS: The upper eyelid island orbicularis oculi myocutaneous flap may be a useful tool for periorbital reconstruction. PMID- 10674733 TI - Involutional entropion and ectropion of the Asian lower eyelid. AB - PURPOSE: A clinical observation showed that involutional entropion of the lower eyelid in Asians may occur more commonly than ectropion. A review of surgical cases was performed to examine this hypothesis. METHODS: A retrospective review of the number of Asian lower lid involutional ectropion and entropion repairs was performed in three different clinical practice settings. These data were compared and statistically analyzed with similar data for non-Asian patients. RESULTS: The frequency of ectropion among Asians was significantly less than in non-Asians (chi-square, p < 0.001). Asian entropion repair represented 11.4% of the 604 eyelid operations performed on Asians, whereas Asian ectropion repair made up only 1.5% of cases. Non-Asian entropion and ectropion repairs were 3.7% and 6.2%, respectively, of the 1,849 eyelid procedures performed on non-Asians. CONCLUSIONS: Because of the normal anteriorly protruding position of the orbital fat within the Asian lower eyelid, Asians may be more predisposed than whites to the development of involutional entropion rather than ectropion. Removal of lower eyelid fat should be considered in entropion repair of the Asian lower eyelid. PMID- 10674734 TI - Lagophthalmos: an unusual manifestation of oculomotor nerve aberrant regeneration. AB - PURPOSE: To describe a patient with unusual findings after regeneration of the oculomotor nerve. METHODS: Case report. RESULTS: A 35-year-old woman developed complete right third nerve paralysis after neurosurgical ligation of internal carotid-posterior communicating and internal carotid-ophthalmic artery aneurysms. Permanent ipsilateral lagophthalmos appeared as third nerve function spontaneously recovered. CONCLUSION: Lagophthalmos may rarely develop after aberrant regeneration of the oculomotor nerve, presumably caused by co contraction of the levator and superior rectus muscles during the Bell's phenomenon. PMID- 10674735 TI - Blepharoptosis and central nervous system abnormalities in combined valproate and hydantoin embryopathy. AB - PURPOSE: To report a case of intrauterine anticonvulsant exposure with subsequent ocular adnexal manifestations. METHODS: Case report. RESULTS: An 18-month-old child with known anticonvulsant embryopathy was referred for the management of bilateral congenital blepharoptosis. Physical examination confirmed ocular and nonocular external manifestations of valproate and hydantoin embryopathies. Cavum septum pellucidum, mild sulcation defects, and cerebellar atrophy were identified on neuroimaging. CONCLUSIONS: To our knowledge, our patient represents the second reported case of anomalous septum pellucidum after intrauterine valproate exposure. Clinicians evaluating patients with craniofacial features associated with intrauterine valproate exposure should recognize that concomitant anomalies of the central nervous system, including the septum pellucidum, might exist. PMID- 10674736 TI - Monocanalicular intubation for dacryostenosis in oculo-auriculo-vertebral dysplasia (hemifacial microsomia) with congenital corneal anesthesia. AB - PURPOSE: To present a case of oculo-auriculo-vertebral dysplasia associated with corneal anesthesia and ipsilateral dacryostenosis that was successfully treated with monocanalicular lacrimal intubation. METHODS: Case report. RESULTS: Previous neurotrophic corneal ulcers in a child with hemifacial microsomia had become secondarily infected from a stagnant tear lake, resulting in significant corneal scarring and visual loss. A single monocanalicular stent maintained nasolacrimal patency without causing further corneal trauma, despite constant medial gaze because of wide lateral tarsorrhaphy and contralateral occlusion therapy for amblyopia. CONCLUSIONS: Monocanalicular stents may aid the treatment of dacryostenosis in the face of compromised corneal sensation. PMID- 10674737 TI - Probing and bicanalicular silicone tube intubation under nasal endoscopy in congenital nasolacrimal duct obstruction. AB - PURPOSE: To evaluate the beneficial effects of nasal endoscopy on preventing complications during probing and bicanalicular silicone intubation, and to determine the structural nasal abnormalities in congenital nasolacrimal duct obstruction. METHODS: Probing and silicone tube intubation under nasal endoscopy were performed in 37 eyes of 26 patients who ranged in age from 7 to 60 months (mean, 18.8 +/- 13.4 months). RESULTS: By nasal endoscopy, the inferior turbinate and meatus appeared normal in 15 patients (58%), whereas 11 patients (42%) had hypertrophy of the inferior turbinate and/or stricture in the inferior meatus. Twenty-four of 26 eyes (92%) were cured by probing only. We performed silicone tube intubation and cured 11 of 12 eyes (92%) including two failures. Of 14 eyes that had undergone failed probings elsewhere, the success rate was 92.8% (13 of 14 eyes). The overall success rate of probing and silicone tube intubation under nasal endoscopy was 97%. CONCLUSION: Nasal endoscopy during probing and bicanalicular silicone tube intubation is useful especially in selected cases of failed probings. Nasal endoscopy should assist the inexperienced surgeon in preventing trauma to the nasal base and septal mucosa, hemorrhage, and passage of the probe under the mucosa rather than through the ostium. PMID- 10674738 TI - Modified orbital decompression for dysthyroid orbitopathy. AB - PURPOSE: The transantral approach to orbital decompression remains useful for the management of exophthalmos associated with dysthyroid orbitopathy. However, the risk of postoperative diplopia is a concern. Preservation of the anterior periorbita may help support the orbital contents and decrease the incidence of diplopia. METHODS: The medical records were reviewed of 15 consecutive patients who underwent 30 transantral orbital decompressions for proptosis associated with dysthyroid orbitopathy. The procedures were completed in standard fashion, including removal of the inferomedial bony strut between the medial orbital wall and the floor. However, stripping of the periorbita was only done posteriorly; the anterior 10 to 15 mm of periorbita were left intact. RESULTS: Six patients had preoperative diplopia that persisted after decompression. Of the nine patients without diplopia preoperatively, none developed diplopia. Proptosis was reduced a mean of 3.5 +/- 2.6 mm. CONCLUSIONS: Preservation of the anterior periorbita during transantral orbital decompression reduces the risk of postoperative diplopia. An adequate reduction in proptosis is also achieved. PMID- 10674739 TI - Absence of seasonal variation in Graves disease. AB - PURPOSE: To determine whether there is an identifiable pattern of seasonal variation in the onset of symptoms of orbitopathy and thyrotoxicosis in patients with Graves disease. METHODS: This retrospective, noncomparative case series reviewed 305 randomly selected office records of patients referred to one author (M.K.) for evaluation of Graves orbitopathy between July 1990 and June 1998. All patients met inclusion criteria for the diagnosis of Graves orbitopathy. During initial evaluation for orbitopathy, patients identified the date of onset of orbital symptoms as well as the earliest date of either onset of thyroid symptoms or documented thyroid abnormality. Patients were excluded from analysis of seasonal variation if they could not recall the month of symptom onset or were euthyroid. The onset of orbital symptoms and identification of dysthyroid state were analyzed by calendar month and season. The chronological relationship of the development of orbital and thyroid symptoms was evaluated. RESULTS: No significant seasonal variation appeared in the onset of orbital symptoms or identification of dysthyroidism. Out of 148 patients, 115 (78%) developed symptoms of orbital disease within 18 months of the identification of dysthyroidism. The most common presenting orbital symptoms were swelling of the lid or prominence of the globe. CONCLUSIONS: This study fails to provide evidence for a seasonal influence on the incidence of Graves disease and the associated orbitopathy. PMID- 10674740 TI - Diagnosis and management of allergic fungal sinusitis with orbital involvement. AB - PURPOSE: Allergic fungal sinusitis (AFS) is a noninvasive disease characterized by recurrent sinusitis. This condition is commonly treated with surgical debridement and several months of systemic corticosteroids. The treatment of AFS is examined in this study. METHODS: A retrospective case series of three patients with AFS. RESULTS: All three patients were treated with surgical debridement and less than one month of systemic corticosteroids. The patients then were treated with intranasal corticosteroids and monitored closely. Antifungal therapy was not used. All three patients remained disease-free during follow-up ranging from 12 months to 36 months. CONCLUSIONS: Surgical debridement and systemic corticosteroids for less than four weeks followed by intranasal corticosteroids may provide long-term control of AFS. Additional study is recommended to examine further the optimal treatment for AFS. PMID- 10674741 TI - Osteogenic sarcoma and phthisis bulbi: a case report. AB - PURPOSE: To describe a case of osteogenic sarcoma (osteosarcoma) that developed within a phthisical eye. METHOD: Case report. RESULTS: An 86-year-old woman with a 20-year history of phthisis bulbi developed pain and proptosis. Tumor was identified by computed tomography. An exenteration was performed, and osteogenic sarcoma was identified. CONCLUSION: Osteogenic sarcoma is the most common primary malignant tumor of bone. In the orbit it frequently is associated with prior irradiation for retinoblastoma. We describe the first case of osteogenic sarcoma that developed de novo from bone within a phthisical eye. PMID- 10674742 TI - Sling removal. PMID- 10674743 TI - Research toward safer resection of the cirrhotic liver. AB - Despite recent advances in hepatic surgery, resection of the cirrhotic liver continues to be fraught with high morbidity and mortality rates. As a result, for many patients requiring resection of HCC the postoperative course is complicated and the probability of cure is diminished by coexisting cirrhosis. In this review, we discuss the characteristics of the cirrhotic liver which make it poorly tolerant of resection and the most common complications that follow such surgery. The main purpose of this paper is to review recent attempts to identify interventions that might be beneficial to cirrhotic patients undergoing resection. These interventions include assessment of liver reserve, advances in surgical technique, and improvement in liver function and regeneration. PMID- 10674744 TI - Cholangiographic features in the diagnosis and management of obstructive icteric type hepatocellular carcinoma. AB - In 11 years and 3 months, 2,037 patients with HCC were seen and 48 patients (2.4%) were diagnosed to have obstructive icteric type HCC. Five patients were terminally ill and were not investigated further. Forty three patients were initially investigated by endoscopic retrograde cholangiography (ERC) or percutaneous transhepatic cholangiogram (PTC) and classified as having obstructive icteric type 1, 2, or 3 HCC based on the cholangiographic findings. The obstruction in type 1 HCC was due to intraluminal tumour casts and/or tumour fragments obstructing the hepatic ductal confluence or common bile duct, while intraluminal blood clots, from haemobilia, filling the biliary tree was the cause in type 2 HCC. The pathology in type 3 HCC was extraluminal obstruction by extensive tumour encasement of the intra-hepatic biliary ductal system and/or extrinsic compression of the hepatic and common bile ducts by tumour(s) and/or malignant lymph nodes. At the initial ERC/PTC, 10 patients (5 resected, 50%) had obstructive icteric type 1 and 23 patients (0 resected) had obstructive icteric type 3 HCC. Of the 10 patients initially classified according to cholangiography to have obstructive icteric type 2 HCC, subsequent investigations revealed that 6 patients had type 1 HCC (4 resectable,67%) and 4 patients had type 3 HCC (0 resectable). The classification of the obstructive icteric type HCC into types 1, 2, and 3, based on the initial cholangiographic appearances has simplified and rationalized our management strategy for this condition. PMID- 10674745 TI - Surgery for deeply located hydatid cysts of the liver: a simple alternative. AB - Gaining access to deeply located hydatid cysts of the liver using conventional surgical techniques may be accompanied by significant intra- and postoperative complications. In addition, obliteration of the cyst cavity is still a matter of controversy. We developed a novel method for easy access to deep hydatid cysts using a water jet dissector (Parenchimotom 01, TOSA, Pleven, Bulgaria). At pressure of 20 Bar using a 0.2 mm nozzle, a corridor is created through the liver parenchyma overlying the cyst; vessels and biliary duct are thus clearly displayed as linear structures traversing the corridor and are ligated and divided under direct visual control. The fibrous capsule of the cyst is spared by the jet. Following endocystectomy performed in the ordinary manner, the cyst cavity is filled with gelatin sponge; a passive tube drain is placed in contact with the liver incision. In allowing for a selective dissection of the liver parenchyma, the jet makes safe access to deeply located hydatid cysts possible. On the other hand, the gelatin sponge induces good fibroblast response and assists in rapid and effective obliteration of the residual cavity. This novel technique works well in our hands but more extensive studies are necessary before its final acceptance. PMID- 10674746 TI - The impairment of wound healing process is correlated with abnormalities of TNF alpha production by peritoneal exudate cells in obstructive jaundiced rats. AB - The wound healing process and production of tumour necrosis factor alpha (TNF alpha) by peritoneal cells of 7-day and 14-day obstructive jaundice (OJ) and sham operated rats were investigated. In the study the skin wound breaking strength was measured. In addition such histological and biochemical parameters as fibroblast and endothelial cell proliferation, inflammatory cell infiltration and hydroxyproline content were evaluated in polyurethane sponge discs implanted subcutaneously into rats. TNF-alpha production by peritoneal exudate cells (PEC), both spontaneous and lipopolysaccharide (LPS)-induced was determined by a bioassay. In OJ rats the process of both early as well as late phase of healing was impaired. The breaking strength of skin wound was decreased, the fibroblast and endothelial cell proliferation and collagen deposition, as well as hydroxyproline content were diminished. In 7 day OJ the numbers of inflammatory cells in the implants were lowered with a subsequent slight increase on day 14 of OJ. The spontaneous and LPS induced TNF-alpha production by PEC were significantly higher in 7 day OJ as compared with sham-operated controls. On day 14 of OJ the LPS-induced TNF-alpha level was, in contrast, much lower and did not differ much from the spontaneous TNF-alpha production. We conclude that the impairment of wound healing in OJ results from disturbances in functioning of the immune system caused by systemic endotoxaemia. PMID- 10674747 TI - Interval laparoscopic cholecystectomy in the management of acute biliary pancreatitis. AB - The timing of laparoscopic cholecystectomy following an attack of acute biliary pancreatitis is controversial. The traditional approach of interval cholecystectomy has been challenged recently. The present study was designed to evaluate the benefits of interval laparoscopic cholecystectomy for patients with mild acute pancreatitis (Ranson less than 3). Nineteen patients with mild pancreatitis underwent ultrasonographic evaluation to confirm the biliary etiology. ERCP was performed in all patients on the first available endoscopy list, and endoscopic sphincterotomy was performed in two patients with calculi or dilated common bile duct on ultrasonographic examination. Medical treatment was administered and laparoscopic cholecystectomy was scheduled after 8-12 weeks to allow the inflammatory process to settle. There were no recurrent attacks of pancreatitis during this period. The degree of difficulty of the laparoscopic procedure was assessed by the presence of adhesions to the gallbladder area, difficulty of dissection in the Calot's triangle, intraoperative bleeding and the need for a drain. Six patients (31.5%) had severe adhesions, difficult dissection of the cystic duct and artery, bleeding and prolonged operating time. In two of these patients (10.5%) the procedure was converted to open cholecystectomy. In conclusion, our results suggest that postponing laparoscopic cholecystectomy in acute pancreatitis patients is not advantageous surgically and does not justify the risk of further morbidity caused by the gallbladder disease. PMID- 10674748 TI - Treatment options for villous adenoma of the ampulla of Vater. AB - INTRODUCTION: Duodenal villous adenoma arising from the ampulla of Vater has a high risk of malignant development. Excluding associated malignant disease prior to resection of an adenoma of the ampulla is not always possible. Therefore, the surgical procedure of choice to treat this rare tumour is still controversial. OBJECTIVE: To evaluate retrospectively results of treatment of villous adenoma arising from ampulla of Vater with dysplasia or associated carcinoma limited to the ampulla. PATIENTS AND METHODS: From 1985 to 1996, eight patients have been diagnosed with ampullary villous adenoma suitable for resection. We have reviewed treatment, morbidity, mortality, follow-up and final outcome. RESULTS: Pancreatoduodenectomy (PD) was performed in 4 patients. Transduodenal ampullectomy and endoscopic resection was performed in 2 patients each. There was no perioperative mortality. None of the patients had biliary, pancreatic or intestinal leakage but two patients who underwent PD had minor postoperative complications. The mean follow-up was 44 (range: 6-132) months. Villous adenoma was associated with adenocarcinoma in 50% of the cases (4/8 patients). During the follow-up both patients who underwent transduodenal ampullectomy developed recurrent disease. All patients initially treated by PD are alive without evidence of recurrent disease. CONCLUSIONS: Treatment of villous adenoma of the ampulla must be individualized within certain limits. In our series, PD achieve good results and it appears to be the procedure of choice in order to treat villous adenomas with proved presence of carcinoma, carcinoma in situ or severe dysplasia. Endoscopic or local resection may be appropriate for small benign tumours in high risk patients. PMID- 10674749 TI - Pancreatic pseudocysts transpapillary and transmural drainage. AB - BACKGROUND: Pancreatic pseudocyst endoscopic drainage has been described as a good treatment option, with morbidity and mortality rates that are lower than surgery. The aim of our study is to describe the efficacy of different forms of endoscopic drainage and estimate pseudocyst recurrence rate after short follow up period. PATIENTS AND METHODS: We studied 30 patients with pancreatic pseudocyst that presented some indication for treatment: persistent abdominal pain, infection or cholestasis. Clinical evaluation was performed with a pain scale, 0 meaning absence of pain and 4 meaning continuous pain. Pseudocysts were first evaluated by abdominal CT scan, and after endoscopic retrograde pancreatography the patients were treated by transpapillary or transmural (cystduodenostomy or cystgastrostomy) drainage. Pseudocyst resolution was documented by serial CT scans. RESULTS: 25/30 patients could be treated. Drainage was successful in 21 (70% in an 'intention to treat' basis). After a mean follow-up of 42 +/- 35.82 weeks, there was only 1 (4.2%) recurrence. A total of 6 complications occurred in 37 procedures (16.2%), and all but 2 were managed clinically and/or endoscopically: there was no mortality related to the procedure. Patients submitted to combined drainage needed more procedures than the other groups. There was no difference in the efficacy when we compared the three different drainage methods. CONCLUSIONS: We concluded that pancreatic pseudocyst endoscopic drainage is possible in most patients, with high success rate and low morbidity. PMID- 10674750 TI - Adenoma of the papillae of Vater. Report of eleven cases. AB - Eleven patients with a preoperative diagnosis of adenoma of the papillae of Vater were followed up during the fifteen-year period from 1984 till 1998 in the Oulu University Hospital. Seven patients were treated primarily by transduodenal excision without any recurrences so far. One of these seven patients was found to have adenocarcinoma in a histological examination. Active surgery for adenoma of the papillae of Vater is recommended because of the precancerous nature of the lesion, and because malignancy cannot always be detected by endoscopic biopsies. Transduodenal excision could be recommend for patients at high operative risk, especially in cases with small adenomas and low-grade dysplasia, where histologically free resection margins can be achieved, but pancreaticoduodenectomy should still be performed on patients at low operative risk. PMID- 10674751 TI - Mucobilia in association with a biliary cystadenocarcinoma of the caudate duct: a rare cause of malignant biliary obstruction. AB - Mucobilia is a rare condition characterized by the accumulation of abundant mucus within the intra- or extrahepatic biliary tree. A variety of hepatobiliary and pancreatic neoplasms are mucin producing and have been associated with the development of mucobilia including biliary mucinosis, biliary papillomatosis, mucin-producing cholangiocarcinoma (MPCC), or cystic neoplasms of the pancreas or biliary tree (cystadenoma or cystadenocarcinoma). We report the case of 46 year old male with a biliary cystadenocarcinoma of the caudate lobe which resulted in chronic biliary obstruction and relapsing cholangitis. A review of the literature for both mucobilia and biliary cystadenocarcinoma is provided along with a discussion addressing the clinical presentation, diagnosis, treatment, and prognosis for this rare entity. PMID- 10674752 TI - Peliosis hepatis with intrahepatic hemorrhage: successful embolization of the hepatic artery. AB - Peliosis hepatis is defined as the appearance of blood filled lakes in the hepatic parenchyma. It has been associated with various pharmacological agents and infections. Treatment has been primarily symptomatic and includes discontinuation of offending medications, partial hepatectomy or occasionally liver transplantation. We report a 58 year old white female on hormone replacement therapy who developed symptomatic peliosis hepatis and underwent successful superselective hepatic artery embolization with control of bleeding. PMID- 10674753 TI - Major HPB procedures must be undertaken in high volume quaternary centres? AB - BACKGROUND: Reports of better results at national referral centers than at low volume community hospitals have prompted calls for regionalizing pancreaticoduodenectomy (the Whipple procedure). We examined the relationship between hospital volume and mortality with this procedure across all US hospitals. METHODS: Using information from the Medicare claims database, we performed a national cohort study of 7,229 Medicare patients more than 65 years old undergoing pancreaticoduodenectomy between 1992 and 1995. We divided the study population into approximate quartiles according to the hospital's average annual volume of pancreaticoduodenectomies in Medicare patients: very low (<1/y), low (1-2/y), medium (2-5/y), and high (5+/y). Using multivariate logistic regression to account for potentially confounding patient characteristics, we examined the association between institutional volume and in-hospital mortality, our primary outcome measure. RESULTS: More than 50% of Medicare patients undergoing pancreaticoduodenectomy received care at hospitals performing fewer than 2 such procedures per year. In-hospital mortality rates at these low- and very-low-volume hospitals were 3- to 4-fold higher than at high-volume hospitals (12% and 16%, respectively, vs. 4%, P<.001). Within the high-volume quartile, the 10 hospitals with the nation's highest volumes had lower mortality rates than the remaining high-volume centers (2.1% vs. 6.2%, P<.01). The strong association between institutional volume and mortality could not be attributed to patient case-mix differences or referral bias. CONCLUSIONS: Although volume-outcome relationships have been reported for many complex surgical procedures, hospital experience is particularly important with pancreaticoduodenectomy. Patients considering this procedure should be given the option of care at a high-volume referral center. PMID- 10674754 TI - Validation of the weight-drop contusion model in rats: a comparative study of human spinal cord injury. AB - Animal models are widely used for studying the pathophysiology as well as treatment strategies for injuries of the central nervous system. However, it is still unclear in how far the rat model of spinal cord injury (SCI) is valid for human SCI. Therefore, comparisons were made among functional, electrophysiological, and morphological outcome parameters following SCI in rats and humans. Contusion of the mid-thoracic spinal cord in 27 adult rats was induced by a weight-drop, leading to severe deficits in open field locomotion at a chronic stage. The data of 85 human patients with chronic SCI were collected and compared with the rat data. In electrophysiological recordings, prolonged latencies and reduced amplitudes in both motor evoked potentials (MEP) and somatosensory evoked potentials (SSEP) were closely correlated to the impairment of locomotor capacity of lower limbs in rats and humans. The morphological parameters assessed by high-resolution magnetic resonance imaging (MRI) in both species indicated that the lesion length and spinal cord atrophy were significantly related to the electrophysiological and functional outcome parameters. In rats, histological analysis was performed and showed, in addition to the MRI, a close relationship between spared white matter and locomotor capacity. Our results suggest an analogous relationship in rats and humans with respect to functional, electrophysiological, and morphological outcomes. Thus, the techniques for evaluating the extent and severity of SCI in humans and rats are of comparable value. This indicates that the rat can serve as an adequate animal model for research on functional and morphological changes after SCI and the effects of new treatment strategies. PMID- 10674755 TI - Neuroprotective effects of gacyclidine after experimental photochemical spinal cord lesion in adult rats: dose-window and time-window effects. AB - The aim of this study was to evaluate the efficacy, optimal dose, and optimal time-window of gacyclidine, a novel N-methyl-D-aspartate (NMDA) receptor antagonist, in terms of its functional, histopathological, and electrophysiological effects after experimental spinal cord injury. The spinal cord of rats was damaged by a photochemical method and the animals were treated by saline or gacyclidine at doses of 1, 2.5, or 5 mg/kg 10 min after injury or gacyclidine 1 mg/kg 10, 30, 60, and 120 min after injury. The time-course of the motor score (walking and inclined-plane stability) was evaluated until day 18, and somatosensory evoked potentials were determined on day 18. The animals were then sacrificed, and the cross-sectional area of the spinal cord (at the epicenter of the injury, above and below the injury) was measured. Walking recovery was better in most of the groups treated after injury than in the untreated injured animals. Motor performances were related to preservation of a larger undamaged area of spinal cord at the level of the injury and, interestingly, with prevention of extension of the anatomical lesion above the level of the injury. Somatosensory evoked potential amplitudes were often higher in treated groups. These results confirm that gacyclidine induces dose-dependent and time-dependent attenuation of spinal cord damage after an experimental vascular lesion. Although all three doses induced neuroprotective effects, recovery was greater and very homogeneous in the group treated with 1 mg/kg. Moreover, recovery was slightly better and more homogeneous within the groups treated 10 and 30 min after injury compared to the other groups. It appears that, according to the existing evidence, NMDA antagonists are an essential component in the elaboration of a neuroprotective strategy after spinal cord trauma. PMID- 10674756 TI - Extracellular N-acetyl-aspartate as a biochemical marker of the severity of neuronal damage following experimental acute traumatic brain injury. AB - We evaluated the acute changes in interstitial and whole brain N-acetyl-aspartate (NAA) measured by high-performance liquid chromatography in animal models of isolated traumatic brain injury (TBI) and TBI combined with secondary insult (hypotension-hypoxia [HH]). The Marmarou impact-acceleration model was used. Four groups were studied: (1) sham-operated control, (2) TBI alone (TBI 500 gm, 2 m), (3) TBI plus 30 min of hypoxia (PaO2, approximately 40 mm Hg) and hypotension (mean arterial blood pressure, approximately 40 mm Hg) (THH), and (4) HH alone. The baseline value for dialysate NAA (NAAd) in the rats was 8.17+/-1 microM. No significant difference between groups was found for this baseline value. The TBI group had a modest (100%) transient increase in NAAd after isolated TBI. The HH group had a transient (500%) increase in NAAd at 1 h, sustained for 2 h. In the THH group, there was a persistent increase in NAAd (800%) that peaked at 2.5 h. The whole brain NAA (NAAw) concentration in controls was 8.5+/-0.5 mmol/kg wet weight. There was no significant difference between TBI and controls; however, there was a significant decrease in NAAw in the THH and HH group compared to controls. Thus, in this animal model of TBI and TBI with secondary insult, we found that persistent, marked elevation in NAA is associated with TBI and secondary ischemic/hypoxic insult, but not with isolated TBI alone. PMID- 10674757 TI - Cerebral hemodynamic effects of 7.2% hypertonic saline in patients with head injury and raised intracranial pressure. AB - The aim of the present study was to investigate the acute effects of 7.2% hypertonic saline (HS) on intracranial pressure (ICP), cerebral and systemic hemodynamics, serum sodium, and osmolality in 14 patients with moderate and severe traumatic brain injury (Glasgow Coma Scale < or =13) and raised ICP (>15 mm Hg) within the first 72 h postinjury. After CO2 reactivity and autoregulation were tested, each patient received a 15-min infusion of 7.2% HS (1,232 mEq/L, volume 1.5 mL/kg). ICP, serial hemodynamics, cerebral blood flow (CBF) estimated from cerebral arteriovenous oxygen content difference (AVDO2), and laboratory variables, including serum osmolality, electrolytes, urea, and creatinine were collected before infusion (T0) and at 5, 30, 60, and 120 min after (T5, T30, T60, T120). Urine output was measured 2 h before infusion and at T120. While CO2 reactivity was preserved in all patients, autoregulation was preserved in only four. ICP decreased to about 30% of base line (p = 0.0001) during the whole study period. During the first hour after infusion, cerebral perfusion pressure (p< or =0.04) and cardiac index (CI; p< or =0.01) increased, while systemic vascular resistance index fell (p< or =0.05). Heart rate increased (p< or =0.04) during the first 30 min. Pulmonary artery occlusion pressure (PAOP) increased (p = 0.004) at T5. There were no significant changes in mean arterial blood pressure (MABP), urine output, and estimated CBF. A significant positive correlation (r = 0.75; p = 0.02) between ICP and serum osmolality was found at T5. The administration of 7.2% HS in patients with traumatic brain injury significantly reduces ICP without significant changes in relative global CBF (expressed as 1/AVDO2), increases CI and transiently increases PAOP, without changing MABP and urine output. The correlation between changes in osmolality and ICP supports the hypothesis that HSS may in part decrease ICP by means of an osmotic mechanism. PMID- 10674758 TI - Characterization of plasma magnesium concentration and oxidative stress following graded traumatic brain injury in humans. AB - Plasma magnesium, calcium, and oxidative status were investigated in 31 male casualties with traumatic brain injury (TBI) during a 7-day posttraumatic period. The study group consisted of eight patients with mild closed head injury (Glasgow Coma Scale score [GCS] of 13-15), 10 patients with extensive penetrating head injury (GCS 4-6), and 13 patients with blast injuries but without direct head trauma. The latter group was included since previous experimental and clinical data have confirmed the development of indirect brain trauma in patients with blast injuries. Patients with multiple injuries were not included. Significant declines in plasma divalent cations were found in GCS 4-6 patients immediately after TBI and persisting for the entire 7-day study period. Similar changes in magnesium, but not calcium, were present in the GCS 13-15 and the blast injury groups, but only up until day 3 after injury. Alterations in lipid peroxidation products and superoxide anions were also observed following TBI. Increased lipid peroxidation was noted in all three groups over the entire posttraumatic period while increases in superoxide anion generation occurred transiently immediately following TBI. Thereafter, in the GCS 13-15 and blast injury groups, superoxide anions subsequently normalized, whereas in extensive head injury (GCS 4-6), superoxide anion generation significantly declined. A negative correlation between magnesium balance and oxidative stress was observed in all patients immediately after injury persisting in GCS 4-6 patients to the end of the observation period. Our findings suggest an interrelationship between magnesium changes and blood oxidants/antioxidants after TBI, which could be of both diagnostic and prognostic value in patients with neurotrauma. PMID- 10674759 TI - Regional expression and role of cyclooxygenase-2 following experimental traumatic brain injury. AB - Prostaglandins, potent mediators of inflammation, are generated from arachidonic acid (AA) via the action of cyclooxygenase-1 and -2 (COX-1 and COX-2). In this study, we report that lateral cortical impact injury in rats significantly increases COX-2 protein levels both in the cortex surrounding the injury site and the ipsilateral hippocampus. COX-2 protein level was elevated as early as 3 h postinjury and persisted for up to 3 days. Increases in immunoreactivity were detected not only in the adjacent cortex and hippocampus, but were also observed in the contralateral cortex and hippocampus, the ipsilateral piriform cortex and the ipsilateral amygdaloid complex. COX-2 immunoreactive cells appear morphologically normal and do not present any of the characteristic features of apoptosis. Double immunostaining experiments using either a neuron-specific or an astroglial-specific marker show that the expression of COX-2 is localized almost exclusively in neuronal cells. Administration of the COX-2 inhibitor 4-[5-(4 methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfona mide (celecoxib, marketed as Celebrex) worsens motor, but not cognitive, performance, suggesting that COX-2 induction following traumatic brain injury may play a protective role. PMID- 10674761 TI - Application of 2,3,5-triphenyltetrazolium chloride staining to evaluate injury volume after controlled cortical impact brain injury: role of brain edema in evolution of injury volume. AB - A reliable method for measuring injury volume after traumatic brain injury (TBI) is of great importance when studying pharmacological protective agents in the field of head trauma research. Utilization of 2,3,5-triphenyltetrazolium chloride (TTC) has gained extensive acceptance in stroke research and has recently been applied to injury volume measurement in the lateral fluid percussion model. The present study was undertaken to apply this method to the controlled cortical impact (CCI) model and to study the role of brain edema. Male Sprague-Dawley rats were subjected to CCI brain injury at a velocity of 3 m/sec and 1 mm (mild), 2 mm (moderate), and 3 mm (severe injury) deformation, while rats in the control group were subjected to the same surgical procedure but received no injury. Absolute and corrected injury volumes with TTC staining and brain edema measurements with the wet-dry method were evaluated at 1, 2, 3, 4, and 7 days after TBI. The most prominent injury volume in the moderate injury group (2 mm deformation) was seen at postinjury day 1 and 2 (day 1, absolute: 49.1+/-5.6, corrected: 40.5+/-7.9; day 2, absolute: 46+/-6.9, corrected: 40.2+/-10.5), whereas the smallest injury volume was found at postinjury day 7 (absolute: 24.9+/-7, corrected: 27.4+/-7.4). The time course of brain edema studies demonstrates that brain edema formation peaks at postinjury day 1. A statistically significant reduction of injury volume was observed after postinjury day 4. We also observed that due to the presence of brain edema absolute injury volume is more than corrected injury volume in the first 3 days after injury as opposed to injury volume at postinjury day 7. These results suggest that the measurement of injury volume with TTC staining should be corrected for brain edema in the CCI brain injury model. PMID- 10674760 TI - The novel compound LOE 908 attenuates acute neuromotor dysfunction but not cognitive impairment or cortical tissue loss following traumatic brain injury in rats. AB - Experimental traumatic brain injury (TBI) initiates massive disturbances in Ca2+ concentrations in the brain that may contribute to neuronal damage. Intracellular Ca2+ may be elevated via influx through voltage-operated cation channels, ligand gated ionotropic channels, and store-operated cation channels (SOCs). In the present study, we evaluated the neurobehavioral and histological effects of acute posttraumatic administration of (R,S)-(3,4-dihydro-6,7-dimethoxy-isoquinoline-1 yl)-2-phenyl-N,N-di[2-(2 ,3,4-trimethoxyphenyl)ethyl]-acetamide (LOE 908), a broad spectrum inhibitor of voltage-operated cation channels and SOCs. Male Sprague-Dawley rats (n = 53) were trained in the Morris water maze, anesthetized (60 mg/kg pentobarbital, i.p.), and subjected to lateral fluid percussion brain injury (2.5-2.7 atm; n = 38) or surgery without injury (n = 15). At 15 min postinjury, animals were randomized to receive intravenous administration of either a high dose of LOE 908 (4 mg/kg bolus followed by 160 mg/kg over 24 h; n = 13), a low dose of LOE 908 (2 mg/kg bolus followed by 80 mg/kg over 24 h; n = 12), or vehicle (n = 13). Uninjured controls received the high dose of LOE 908 (n = 8) or vehicle (n = 7). Treatment with either dose of LOE 908 significantly improved neuromotor function at 48 h postinjury when compared to vehicle treatment. Although a significant deficit in visuospatial memory was observed in brain-injured animals at this timepoint when compared to uninjured animals, neither dose of LOE 908 attenuated injury-induced cognitive dysfunction. Histological evaluation revealed that neither dose of LOE 908 affected cortical lesion size at 48 h postinjury. These data suggest that broad spectrum cation channel blockers may be beneficial in the treatment of neurological motor dysfunction when administered in the acute posttraumatic period. PMID- 10674762 TI - Fluid percussion injury transiently increases then decreases brain oxygen consumption in the rat. AB - The oxygen consumption (VO2 microL/h/mg) of sham and of traumatized rat brains within 30 min and 6 h after a lateral fluid percussion injury (FPI) was measured with the Cartesian microrespirometer. Brain slices were cut at the plain of injury and site-specific 20-60-microg cores of tissue were transferred to the microrespirometer. In sham brains, the cortical VO2 (CVO2) was 13.78+/-0.64 and the hippocampal VO2 (HPVO2) was 11.20+/-0.58 microL/h/mg (p<0.05). Within 30 min of the injury, the respective values of 16.89+/-0.55 and 14.91+/-0.06 were significantly increased (p<0.05). The combined VO2 (CVO2, HPVO2) of 12.49+/-0.06 microL/h/mg in shams was significantly less than the combined VO2 of 15.90+/-0.59 microL/h/mg at 30 min post FPI (p<0.001). The maximal CVO2 of 19.49+/-1.10 microL/h/mg and the maximal HPVO2 of 15.98+/-0.99 microL/h/mg were both obtained from the ipsilateral side of the injury. Whereas the contralateral cortical value for injured brains was not significantly different from that of the shams, both ipsilateral and contralateral hippocampal values were significantly greater than that of the shams in response to injury (p<0.05). By 6 h postinjury, the combined VO2 had dropped to 10.01+/-0.84 microL/h/mg but was not significantly lower than the sham values. The data indicate that normal CVO2 is greater than normal HPVO2. The FPI produces significant increases in both CVO2 and HPVO2. Also, while the immediate increase in CVO2 appears to be injury-site dependent, that is, regional, the increase in HPVO2 appears to be global. PMID- 10674763 TI - FDA handling of ADR reports needs improvement. PMID- 10674764 TI - Non-patient-care activities dilute pharmacists' time, NACDS study shows. National Association of Chain Drug Stores. PMID- 10674765 TI - Pharmaceuticals hit the internet auction block. PMID- 10674766 TI - Oral and i.v. formulations of gatifloxacin cleared for U.S. market. PMID- 10674768 TI - Status report on VA national formulary issued. PMID- 10674767 TI - Topical treatment approved for fungal nail infection. PMID- 10674769 TI - Precautions urged in disposing of prescription containers. PMID- 10674770 TI - Researchers find differences in effectiveness of ADHD treatment strategies. PMID- 10674771 TI - BPS roster of certified pharmacists nears 3000 mark. PMID- 10674772 TI - Study provides additional support for hypericum extract. PMID- 10674773 TI - Inadvertent intrauterine infusion of ampicillin-sulbactam. PMID- 10674774 TI - Identifying noncompliance by combining refill audits with telephone follow-up. PMID- 10674775 TI - Aspirin and dipyridamole for stroke prevention. PMID- 10674776 TI - New drugs for the treatment of rheumatoid arthritis. AB - New pharmacologic treatment options for rheumatoid arthritis (RA) are described. Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely prescribed for RA but are limited by the risk of adverse effects, especially gastrointestinal and renal toxicity. The therapeutic effects of these agents are mediated primarily through inhibition of cyclooxygenase (COX) and prevention of subsequent formation of prostaglandins and related inflammatory mediators. Nonspecific COX inhibition appears to be responsible for much of the toxicity of NSAIDs. Agents have been developed that can selectively inhibit the COX-2 isoform, while sparing COX-1. Celecoxib and other COX-2 inhibitors appear to be no more efficacious than conventional NSAIDs, but offer superior safety. COX-2 inhibitors should be considered for patients who are candidates for NSAID therapy but at risk for GI bleeding. Unlike disease-modifying antirheumatic drugs (DMARDs), these agents do not alter underlying disease progression. Leflunomide is a newer DMARD that reduces pyrimidine synthesis, thus decreasing rheumatoid inflammation. Leflunomide appears to be as effective as methotrexate but, unlike that drug, does not necessitate monitoring for bone marrow toxicity. Etanercept, the first biological agent with FDA-approved labeling for use in RA, has shown efficacy and minimal toxicity, except for injection-site reactions. Other biologicals that have been investigated for use in RA include infliximab and interleukin-1 receptor antagonist. COX-2 inhibitors, leflunomide, and etanercept are promising new drugs available for treating RA. Other agents are under development. PMID- 10674777 TI - Atypical antipsychotic agents: a critical review. AB - The pharmacology, efficacy, and adverse effects of atypical antipsychotic agents when used to treat schizophrenia and other disorders are reviewed. Atypical antipsychotic agents were developed in response to problems with typical agents, including lack of efficacy in some patients, lack of improvement in negative symptoms, and troublesome adverse effects, especially extrapyramidal symptoms (EPSs) and tardive dyskinesia CTD). Atypical antipsychotics differ from typical psychotics in their "limbic-specific" dopamine type 2 (D2)-receptor binding and high ratio of serotonin type 2 (5-HT2)-receptor binding to D2 binding. In clinical trials in patients with non-treatment-resistant schizophrenia, risperidone and olanzapine were superior to placebo for positive and negative symptoms. Risperidone and olanzapine were superior to haloperidol on some measures. Quetiapine was better than placebo but was not better than typical antipsychotics. Head-to-head comparisons of atypical antipsychotics in non treatment-resistant schizophrenia have been inconclusive. Clozapine remains the standard agent for treatment-resistant schizophrenia. Atypical agents are substantially more expensive than their typical antipsychotic counterparts. To fully determine the overall efficiency of these drugs, other potential benefits, such as improved quality of life, need to be factored in. Atypical antipsychotics are associated with a decreased capacity to cause EPSs, TD, neuroleptic malignant syndrome, and hyperprolactinemia. Clozapine carries a risk of agranulocytosis; the white blood cell count must be monitored. Atypical antipsychotics are increasingly being used for indications other than schizophrenia, such as the management of aggression, mania, and depression. Atypical antipsychotics are often considered first-line agents for treating schizophrenia and are promising treatment alternatives for other psychiatric and neurologic conditions. PMID- 10674778 TI - Respiratory syncytial virus (RSV) immune globulin and palivizumab for prevention of RSV infection. AB - The efficacy, safety, administration, and advantages and disadvantages of respiratory syncytial virus (RSV) immune globulin and palivizumab for preventing RSV infection are discussed. Prevention of RSV infection has attracted considerable attention because of its dinical and economic impact. Studies have shown respiratory syncytial virus immune globulin intravenous (RSV-IGIV) and palivizumab to be effective in decreasing the number of hospitalizations and hospital days attributable to RSV. The number of intensive-care-unit admissions and the severity of RSV infection in high-risk children decreased with the use of these agents. Both agents have been well tolerated, with few adverse effects; however, their high cost necessitates strict guidelines on use. The patient populations at greatest risk are those with bronchopulmonary dysplasia, those with congenital heart disease, those with a history of apnea or respiratory arrest, immunocompromised patients, those with pulmonary consolidation on chest radiography, and those born prematurely. American Academy of Pediatrics guidelines do not preferentially recommend use of either agent; each has advantages and disadvantages. Prophylactic therapy with RSV-IGIV or palivizumab may reduce the likelihood of RSV infection in high-risk patients. PMID- 10674780 TI - Preparing for a joint commission survey: a pharmacist's perspective. PMID- 10674779 TI - Outcomes of an antimicrobial control program in a teaching hospital. AB - The clinical outcomes and cost-effectiveness of an antimicrobial control program (ACP) were studied. The impact of an ACP in a teaching hospital was analyzed by comparing clinical outcomes and intravenous antimicrobial costs over two two-year periods, the two years before the program and the first two years after the program's inception. Admission baseline data, length of stay, mortality, and readmission rates were gathered for each patient. Patients were identified by using the International Classification of Diseases. Multivariate logistic regression models were constructed for mortality and for lengths of stay of 12 or more days. The acquisition costs of intravenous antimicrobial agents for the second baseline year and the entire program period were tabulated and compared. The average daily inpatient census was determined. The ACP was associated with a 2.4-day decrease in length of stay and a reduction in mortality from 8.28% to 6.61%. Rates of readmission for infection within 30 days of discharge remained about the same. Inpatient pharmacy costs other than intravenous antimicrobials decreased an average of only 5.7% over the two program years, but the acquisition cost of intravenous antimicrobials for both program years yielded a total cost saving of $291,885, a reduction of 30.8%. The institution's average daily census fell 19% between the second baseline year and the second program year. An ACP directed by a clinical pharmacist trained in infectious diseases was associated with improvements in inpatient length of stay and mortality. The ACP decreased intravenous antimicrobial costs and facilitated the approval process for restricted and nonformulary antimicrobial agents. PMID- 10674781 TI - Clinical clerkships for Japanese graduate pharmacy students in U.S. medical centers. PMID- 10674782 TI - Oseltamivir phosphate. PMID- 10674783 TI - Teleradiology in Europe. AB - The new concept of teleradiology is centered on the consideration that it involves management of medical information rather than the simple transmission of diagnostic images from one location to another. Teleradiology must therefore be able to contribute to the seamless integration of the digital environment in which medical data are managed throughout and beyond the hospital, generating value added services for the patients as well as prospective economical benefits for the institution. In this perspective the evolution of telecommunication with the spectacular success of mobile telephony and Internet will play and increasingly important role, by allowing further development in the exchange of multimedia medical information on a regional as well as international level. However, new responsibilities are being given to the radiologists, who must take all necessary technical and organizational actions in order to avoid that the digital management of data may endanger the confidentiality and the integrity of patients' data. PMID- 10674784 TI - Management of change in health care organisations and human resource role. AB - The paper is focused on the analysis of the most relevant factors necessary to manage change in health care organisations. The approach suggested is the Stakeholder one. According to this approach, the hospital's managers seem to be successful if they are able to satisfy people (internal and external stakeholders) that have a stake in the health care institution. The attention of the author is mainly focused on the internal forces that make the health care sector competitive and successful. In order to motivate internal human resources to accept change and to achieve the organisational targets two main methods can be suggested. The former is based on tangible variables and in particular on a fair reward system; the latter is built on intangible elements e.g. communication, negotiation, contracting, and organisational values sharing. Moreover, in order to cope with change it is important to develop the information technology management and to reengineer delivery processes, taking into consideration both the costs and benefits of these kinds of innovations. PMID- 10674785 TI - Organ doses, detriment and genetic risk from interventional vascular procedures in Malaga (Spain). AB - Nowadays, the radiological risk from simple X-ray procedures is well known. The purpose of this work has been to estimate the population risk from digital angiographic and interventional procedures and to compare it with the one from simple procedures in the same population. The population risk has been estimated according to the following quantities: genetically significant dose, somatic significant dose, collective effective dose, annual per caput effective dose and detriment. These have been estimated from dose area product and organ dose. Organ dose values were estimated with the Eff-Dose software. A population of 605410 people were included in the study. In 1996, 1312 patients were to digital interventional vascular procedures in Malaga, and 159 of them were selected in this research project to obtain the dose area product and organ dose. The results obtained for the quantities evaluated are: genetically significant dose, 4.1 microGy; somatic significant dose, 0.9 mSv; collective effective dose, 11.65 person-Sv: annual per caput effective dose, 0.02 mSv and detriment, 0.65 radiogenic cancers per year. These procedures supply a high radiation dose, so they should have a greater contribution to population dose and risk than simple examinations. However, our results indicate just the opposite. PMID- 10674786 TI - A pilot study on the quality control of film processing in medical radiology laboratories in Greece. AB - The results of a pilot study on the quality of film processing in 80 medical diagnostic radiology laboratories all over Greece are presented. The sensitometric technique for the evaluation of processing has been used to calculate film's base + fog, maximum optical density, speed and contrast, parameters which describe the performance characteristics of automatic film processors and films. The mean values of the base + fog and the maximum optical density were well within the acceptance limits. The film speed was almost constant while the film contrast showed significant variation. PMID- 10674787 TI - Breast cancer screening; cost-effective in practice? AB - The main aim of national breast screening is a reduction in breast cancer mortality. The data on the reduction in breast cancer mortality from three (of the five) Swedish trials in particular gave rise to the expectation that the Dutch programme of 2-yearly screening for women aged 50-70 would produce a 16% reduction in the total population. In all likelihood, many of the years of life gained as a result of screening are enjoyed in good health. According to its critics the actual benefit that can be achieved from the national breast cancer screening programmes is overstated. Considerable benefits have recently been demonstrated in England and Wales. However, the fall was so considerable in such a relatively short space of time that screening (started in 1987) was thought to only have played a small part. As far as the Dutch screening programme is concerned it is still too early to reach any conclusions about a possible reduction in mortality. The first short-term results of the screening are favourable and as good as (or better than) expectations. In Swedish regions where mammographic screening was introduced, a 19% reduction in breast cancer mortality can be estimated at population level, and recently a 20% reduction was presented in the UK. In countries where women are expected to make appointments for screening themselves, the attendance figures are significantly lower and the quality of the process as a whole is sometimes poorer. The benefits of breast cancer screening need to be carefully balanced against the burden to women and to the health care system. Mass breast screening requires many resources and will be a costly service. Cost-effectiveness of a breast cancer screening programme can be estimated using a computer model. Published cost-effectiveness ratios may differ tremendously, but are often the result of different types of calculation, time periods considered, including or excluding downstream cost. The approach of simulation and estimation is here the same for all countries. The effects of a breast-screening program depend on many factors, such as the epidemiology of the disease, the health care system, costs of health care, the quality of the screening programme and the attendance rate. The estimated CE-ratio ranges from 2650 euros per life-year gained in Navarra to 9650 in Germany. Although relatively low incidence levels expected, the CE-ratio in Navarra is most favourable probably due to a relatively unfavourable clinical stage distribution before screening and the increasing incidence. The UK has a screening situation that is almost similar with the Netherlands. Therefore, the CE-ratios of both countries are comparable. The differences between countries make it impossible to set up one uniform screening policy. The theoretical outcomes of the benefit that can be achieved are generally from small-scale trials involving a limited number of experts, persons examined, and areas. On a national scale, with hundreds of professional practitioners, it can be expected to be more difficult to attain uniform quality. Continuous quality control, monitoring and evaluation are therefore crucial. PMID- 10674788 TI - Dolichoodontoid. A rare cranio-cervical anomaly--MRI findings. AB - The case of a 40-year-old woman with a dolichoodontoid, a rare congenital anomaly of the cranio-cervical region, is presented. Due to summation image and overlying bony structures, plain radiographs in two planes were inconclusive. MRI revealed the hyperplasia of the odontoid process, allowed a grading of the subtype of this disorder and demonstrated its relationship to the neural structures within the foramen magnum and the upper cervical spine. Additional inflammatory disease, suspected in this patient with long standing rheumatoid arthritis could be excluded by MRI. PMID- 10674789 TI - 3D MR angiography of the entire aorta: modified application of the body-phased array coil for a single-shot technique. AB - OBJECTIVE: Evaluation of different contrast-enhanced MR angiography imaging protocols for visualization of the entire aorta in breath-hold technique. METHODS AND PATIENTS: Three different CE (0.15 mmol/kg) MRA protocols were evaluated by phantom and patient studies: (1) two separate MRA with conventional application of the body-phased array coil; (2) a single-shot MRA with modified application of the body-phased array coil; (3) a single-shot MRA with the body coil. Duplex sonography, CTA and DSA were used as standard of reference. RESULTS: In all examinations the entire aorta could be visualized. The best SNR was acquired with protocol (1). The SNR of protocol (2) was reduced if the sagittal body diameter of the patient was greater than 20 cm and decreased significantly with diameters over 30 cm. By the use of protocol (3) the SNR was notably poor. The quality scored for the visualization of the entire aorta was 97.5% (protocol 1); 92.5% (protocol 2); and 80.0% (protocol 3). CONCLUSION: In most cases the modified application of the body-phased array coil allows the imaging of the entire aorta as a single-shot 3D CE MRA in diagnostic quality. PMID- 10674790 TI - Real-time interactive virtual endoscopy of the tracheo-bronchial system: influence of CT imaging protocols and observer ability. AB - OBJECTIVE: To evaluate the influence of different spiral CT examination protocols suitable for clinical use on image quality and to assess the observer dependence in interactive real-time virtual bronchoscopy. METHODS AND PATIENTS: Real-time perspective volume rendering of the airways in twenty normal patients based on four different spiral CT examination protocols was evaluated by four observers in regard to the order of depictable bronchi. RESULTS: Best results were obtained using an examination protocol with a small beam collimation and a maximum pitch. Depending on the observer's ability to control the fly path and the orientation of the bronchi with respect to the slice plane up to sixth order bronchi could be depicted. Inter-observer variability was up to two branching orders. CONCLUSION: The performance of virtual bronchoscopy strongly depends on the applied CT examination protocol and the observers experience with perspective volume rendering. Both of which have to be taken into account when virtual bronchoscopy is compared with fiberoptic bronchoscopy. PMID- 10674791 TI - Solitary subependymal giant cell astrocytoma: case report. AB - In this report, we describe a case of subependymal giant cell astrocytoma in a patient lacking clinical symptoms of tuberous sclerosis. The absence of any features of tuberous sclerosis initially dissuaded us from including subependymal giant cell astrocytoma in our differential diagnosis. PMID- 10674792 TI - Persistent hypoglossal artery associated with arteriovenous malformation: a case report. AB - A rare case of persistent hypoglossal artery in conjunction with arteriovenous malformation was presented. MRA could delineate persistent hypoglossal artery and arteriovenous malformation very clearly. The patient suffered from intracranial hemorrhage from in the 37th week of pregnancy. MRI, MRA, angiography, and CT of this rare condition are reported. PMID- 10674793 TI - Laryngopyocele: signs on computed tomography. AB - A laryngocele is an air-filled dilation of the saccule of the larynx. An infected laryngocele is called a laryngopyocele. Our experience with a case of laryngopyocele with signs on computed tomography before and after antibiotic therapy is presented since laryngopyocele is more unusual. PMID- 10674794 TI - Latent learning in medial temporal amnesia: evidence for disrupted representational but preserved attentional processes. AB - Damage to the hippocampus and medial temporal (MT) structures can lead to anterograde amnesia and may also impair latent learning, in which prior exposure to cues affects their subsequent associability. Normally, latent learning may reflect both representational and attentional mechanisms. Prior work has suggested that individuals with MT amnesia have specific deficits in representational processing; thus, latent learning that invokes primarily representational mechanisms might be especially impaired in MT amnesia. The current results provide preliminary confirmation of this prediction. In Experiment 1, a latent learning paradigm expected to invoke representational mechanisms was impaired in individuals with MT amnesia, whereas in Experiment 2, a paradigm expected to invoke other attentional mechanisms was spared in individuals with MT amnesia. This suggests the representational and attentional components of latent learning are dissociable and differentially affected in anterograde amnesia. PMID- 10674795 TI - Goal-directed selective attention and response competition monitoring: evidence from unilateral parietal and anterior cingulate lesions. AB - Competing visual stimuli lead to slower responses to targets. This response competition must be resolved before correct responses are executed. Neuroimaging suggests that response competition monitoring may be subserved by an integrated neural network including the anterior cingulate cortex (ACC). In this study, 1 patient with a parietal lesion (Patient J.S.) and 1 with an ACC lesion (Patient G.M.) were presented with 2 flanker tasks; 1 required verbal identification of color targets, and the other required an opposite response to targets (e.g., see red and say "green"); a control group was also tested. For controls, perceptually incongruent flankers interfered with the ability to inhibit prepotent responses to targets. Patient J.S. performed in a similar manner, even when flankers appeared in the neglected field. Patient G.M. demonstrated reduced interference effects for contralesional flankers. Results are discussed in terms of goal directed selective attention and response competition monitoring. PMID- 10674796 TI - Structural brain correlates of verbal and nonverbal fluency measures in Alzheimer's disease. AB - This study examined the relationships between regional brain volumes and semantic, phonological, and nonverbal fluency in 32 participants with Alzheimer's disease (AD). Object but not animal semantic fluency correlated with frontal and temporal gray matter volumes. Phonological fluency was not significantly associated with any brain volume examined. Nonverbal fluency was selectively associated with bilateral frontal gray matter volumes. Hippocampal volumes, although markedly reduced in these patients, were not related to any of the fluency measures. Results lend evidence to the importance of the frontal lobes in the directed generation of nonverbal and verbal exemplars by AD patients. Furthermore, both left- and right-hemisphere regions contribute to the generation of verbal and nonverbal exemplars. PMID- 10674797 TI - The cerebral hemispheres cooperate to perform complex but not simple tasks. AB - Three experiments were designed to examine whether task complexity determines the degree to which a division of processing across the hemispheres (i.e., across hemisphere processing) underlies performance when within- and across-hemisphere processing are equally possible. When task complexity was relatively low, performance in a midline condition that allowed for either within- or across hemispheric processing resembled within-hemisphere performance (Experiments 1 and 2). However, when task complexity was high, performance in a midline condition (Experiments 1 and 2) and a lateralized condition, which also allowed for either within- or across-hemisphere processing (Experiment 3), resembled across hemisphere performance. Results complement and extend prior work (e.g., M. T. Banich & A. Belger, 1990) by indicating that the degree to which interhemispheric cooperation underlies performance changes with the complexity of the task being performed. This finding suggests that the hemispheres dynamically couple or uncouple their processing as a function of task complexity. PMID- 10674798 TI - Interhemispheric visual integration in three cases of familial callosal agenesis. AB - Three cases of callosal agenesis (a 39-year-old woman and her 11- and 12-year-old daughters) were tested on their ability to integrate visual information between the visual hemifields. They were all able to name colors and digits in either hemifield with high accuracy and were able to decide whether letters or digits in opposite hemifields were the same or different. They had greater difficulty deciding whether colors in opposite hemifields were the same or different. When shown 6-letter words made up of pairs of 3-letter words that straddled the midline (e.g., MANAGE, ROTATE), they responded to them as whole words and never as 3-letter words, suggesting perceptual continuity across the midline, at least for verbal material. The most likely interpretation is that the integration of form, but not color, is achieved through the intact anterior commissure in these participants. PMID- 10674799 TI - Salivary testosterone concentrations in left-handers: an association with cerebral language lateralization? AB - The level of testosterone exposure in early brain development may influence the direction or degree of cerebral language lateralization. Possible links between individual differences in testosterone levels and patterns of speech representation were investigated in 180 healthy young adults (97 left handed, 83 right handed) using the Fused Dichotic Words Test (T. Halwes, 1991). Among left handed participants, significantly higher testosterone concentrations were observed in individuals with a left-ear advantage on dichotic listening than in individuals with a right-ear advantage. Among right-handed participants, the pattern of group differences in testosterone tended to be reversed, resulting in a statistically significant interaction. Results extend prior findings by S. D. Moffat and E. Hampson (1996a) and raise the possibility that higher testosterone is associated with patterns of brain organization in which speech and praxic functions are lateralized to the same hemisphere. PMID- 10674800 TI - Memory enhancement for emotional stimuli is impaired in early Alzheimer's disease. AB - Emotional arousal is associated with enhanced memory in neurologically intact individuals, but it is unknown whether this effect is obtained in Alzheimer's disease (AD). The current study compared emotional memory and emotional reactions in patients with early AD and in older controls. Participants viewed emotionally arousing (both pleasant and unpleasant) and neutral photographs while cognitive and electrophysiological reactions were assessed. Memory was tested by free recall and recognition. Emotional reactions were normal in the AD group, but the emotional memory effect (enhanced memory for emotional vs. neutral stimuli) was impaired. Recall results indicated that this effect was normal for pleasant stimuli but abnormal for unpleasant stimuli. These results suggest that the neural basis for the emotional memory effect may be disrupted in AD. Findings are discussed in terms of the role of the amygdala in mediating emotional memory. PMID- 10674801 TI - Age differences in explicit and implicit memory for pictures. AB - In the present study, the authors examined age effects in memory for nonverbal material. A picture fragment completion task was used to test explicit and implicit memory in a younger and an older group. Explicit memory was indexed by free recall of pictures, whereas implicit memory was indexed by perceptual learning (priming). Both free recall and perceptual learning performance were found to be impaired in the older group. A measure of executive functioning was found to be predictive of both explicit and implicit memory. PMID- 10674802 TI - Effects of aging on efficiency of task switching in a variant of the trail making test. AB - The Trail Making Test (TMT; R. M. Reitan, 1958, 1992) is extensively used in research in neuropsychology and in aging, in part because it has been postulated to reflect executive processes, such as planning and switching. However, neurocognitive and individual-difference-based analyses of this test are complicated because of different spatial arrangements of targets, the use of letters only in Version B, and potential order effects when Version A is administered prior to Version B. The present article examines a variant of a TMT (called the Connections Test) that attempts to remedy these deficiencies. A structural equation model suggested that there were no direct effects of age on either the nonalternating or alternating versions of the Connections Test (analogous to TMT Versions A and B, respectively); rather, all age-related effects were mediated through effects on perceptual speed. Moreover, although the nonalternating and alternating versions were strongly related to one another, only the latter had significant independent relations with measures of higher order cognition. PMID- 10674803 TI - Verbal pragmatics following unilateral stroke: emotional content and valence. AB - Verbal pragmatic aspects of discourse production were examined in 16 right brain damaged (RBD), 16 left brain-damaged (LBD), and 16 normal control right-handed adults. The facilitation effect of emotional content, valence hypothesis, and relationship between pragmatics and emotion were evaluated. Participants produced monologues while recollecting emotional and nonemotional experiences. Transcribed monologues were rated for appropriateness on 6 pragmatic features: conciseness, lexical selection, quantity, relevancy, specificity, and topic maintenance. Overall, brain-damaged groups were rated as significantly less appropriate than normals. Consistent with the facilitation effect, emotional content enhanced pragmatic performance of LBD aphasic participants yet suppressed performance of RBD participants. Contrary to the valence hypothesis, RBD participants were more impaired for positive emotions and LBD participants for negative emotions. Pragmatic appropriateness was not strongly correlated with a measure of emotional intensity. PMID- 10674804 TI - The impact of positive and negative feedback on reaction time in brain-damaged patients. AB - Little is known about the impact of feedback on the reaction times (RTs) of brain damaged (BD) patients. The authors therefore investigated the effect of positive and negative feedback on these patients, using a 4-choice RT task. Participants were 107 BD patients with different etiologies and 50 orthopedic (OG) control patients. Patients were assigned to 3 groups in which performance-independent negative, positive, and no feedback were given. Statistical analysis showed that negative feedback led to significantly shorter RTs in BD patients. Even BD patients with high depression scores were affected by negative feedback. In contrast, negative feedback had no impact on the RTs of the OG controls, and positive feedback had no influence on the RTs of any group. These results raise some interesting questions about motivational processes in BD patients. PMID- 10674805 TI - Inhibitory tagging in inhibition of return is affected in schizophrenia: evidence from the stroop task. AB - L. J. Fuentes, A. B. Vivas, and G. W. Humphreys (1999b) showed that stimulus processing is affected when stimuli are presented to locations subject to inhibition of return. They argued that activated representations of stimuli presented at inhibited locations are disconnected from their associated responses through an "inhibitory tagging" mechanism occurring in inhibition of return. In the present research, the authors asked whether such a mechanism is affected in people with schizophrenia. Healthy adults and patients with schizophrenia performed a Stroop task in an inhibition of return paradigm. Healthy adults showed a reduction in the Stroop interference when stimuli were presented at inhibited locations, a result that agrees with the inhibitory tagging mechanism hypothesis and replicates previous findings. However, patients with schizophrenia did not show such a reduction, a result suggesting that they have a deficit in inhibitory processing occurring in inhibition of return. PMID- 10674806 TI - Strategic processing and episodic memory impairment in obsessive compulsive disorder. AB - There is evidence that nonverbal memory problems in obsessive compulsive disorder (OCD) are mediated by impaired strategic processing. Although many studies have found verbal memory to be normal in OCD, these studies did not use tests designed to stress organizational strategies. This study examined verbal and nonverbal memory performance in 33 OCD patients and 30 normal control participants with the Rey-Osterrieth Complex Figure Test and the California Verbal Learning Test. OCD patients were impaired on verbal and nonverbal measures of organizational strategy and free recall. Multiple regression modeling indicated that free recall problems in OCD were mediated by impaired organizational strategies used during learning trials. Therefore, verbal and nonverbal episodic memory deficits in OCD are affected by impaired strategic processing. Results are consistent with neurobiological models proposing frontal-striatal system dysfunction in OCD. PMID- 10674807 TI - Review: tissue engineering for regeneration of articular cartilage. AB - Joint pain due to cartilage degeneration is a serious problem, affecting people of all ages. Although many techniques, often surgical, are currently employed to treat this affliction, none have had complete success. Recent advances in biology and materials science have pushed tissue engineering to the forefront of new cartilage repair techniques. This review seeks to condense information for the biomaterialist interested in developing materials for this application. Articular cartilage anatomy, types of injury, and current repair methods are explained. The need for biomaterials, current commonly used materials for tissue-engineered cartilage, and considerations in scale-up of cell-biomaterial constructs are summarized. PMID- 10674808 TI - Surface-modification of polystyrene-microtitre plates via grafting of glycidylmethacrylate and coating of poly-glycidylmethacrylate. AB - Photografting of glycidylmethacrylate (GMA) onto commercially available polystyrene-microtitre plates was carried out in methanol as well as butanol with benzophenone (BP) as photoinitiator. Alternatively, prepolymers of polyglycidylmethacrylate (PGMA) were synthesized in methanol with azo-iso butyrodinitrile (AIBN) as initiator and dip-coated onto polystyrene-microtitre plates. Both modification methods were tested in order to reach a high binding capacity for proteins. Peroxidase was used as model protein. In addition, the immobilization of myelin basic protein (MBP) to epoxy-modified microtitre plates is shown and a MBP-based ELISA has been developed. PMID- 10674809 TI - Titanium containing amorphous hydrogenated carbon films (a-C: H/Ti): surface analysis and evaluation of cellular reactions using bone marrow cell cultures in vitro. AB - Amorphous hydrogenated carbon (a-C : H) coatings, also called diamond-like carbon (DLC), have many properties required for a protective coating material in biomedical applications. The purpose of this study is to evaluate a new surface coating for bone-related implants by combining the hardness and inertness of a-C : H films with the biological acceptance of titanium. For this purpose, different amounts of titanium were incorporated into a-C : H films by a combined radio frequency (rf) and magnetron sputtering set-up. The X-ray photoelectron spectroscopy (XPS) of air-exposed a-C : H/titanium (a-C : H/Ti) films revealed that the films were composed of TiO2 and TiC embedded in and connected to an a-C : H matrix. Cell culture tests using primary adult rat bone marrow cell cultures (BMC) were performed to determine effects on cell number and on osteoblast and osteoclast differentiation. By adding titanium to the carbon matrix, cellular reactions such as increased proliferation and reduced osteoclast-like cell activity could be obtained, while these reactions were not seen on pure a-C : H films and on glass control samples. In summary, a-C : H/Ti could be a valuable coating for bone implants, by supporting bone cell proliferation while reducing osteoclast-like cell activation. PMID- 10674810 TI - Tissue responses to natural aragonite (Margaritifera shell) implants in vivo. AB - The purpose of this study was to access tissue reactions to the outer prismatic (prism) and the inner nacreous (nacre) layers of the fresh water Margaritifera shell. The materials, in granule form, were implanted into the back muscles and femurs of rats for 1, 2, 4, 8 and 16 weeks. In the back muscles, a foreign body reaction was observed around the implants, starting from one week after implantation and reaching maximal proportions at two weeks. After four weeks, a thin layer of fibrous tissue encapsulated the implanted particles. The external surface of the material stained strongly with acid fuchsin, indicating degradation of implant. At femoral sites, newly formed bone was directly applied to the implant surfaces. The outer-most parts of the organic sheets in prisms were not degraded until 16 weeks after implantation and were embedded in the newly formed bone. The interface between bone and the implants showed close fusion by scanning electron microscopy (SEM). Energy dispersive X-ray analysis (EDAX) demonstrated a phosphorous-rich zone in the interface between bone and the implants, and no electron-dense layer in the interface was found by transmission electron microscopy (TEM). We conclude that Margaritifera shells are biocompatible, biodegradable and osteoconductive materials. Bonding between this natural aragonite and bone seems to occur via a phosphorous-rich intermediate layer. PMID- 10674811 TI - Ion-beam-assisted deposition (IBAD) of hydroxyapatite coating layer on Ti-based metal substrate. AB - A hydroxyapatite layer was formed on the surface of a Ti-based alloy by ion-beam assisted deposition. The deposition methodology comprised of an electron beam vaporizing a pure hydroxyapatite target, while an Ar ion beam was focused on the metal substrate to assist deposition. All deposited layers were amorphous, regardless of the current level of the ion beam. The bond strength between the layer and the substrate increased steadily with increasing current, while the dissolution rate in a physiological saline solution decreased remarkably. These improvements were attributed to an increase in the Ca/P ratio of the layer. Without ion beam assistance, the Ca/P ratio was much lower than the stoichiometric HAp (Ca/P = 1.67). With ion-beam assistance, the Ca/P ratio of the layer increased presumably due to the high sputtering rate of P compared to that of Ca from the layer being coated. PMID- 10674812 TI - Fluoride release profiles of mature restorative glass ionomer cements after fluoride application. AB - This study investigates the fluoridation of four conventional glass ionomer cements (GIC) (ChemFil Superior encapsulated, Fuji Cap II, Ketac-Fil and Hi Dense) and three resin-modified GIC (RM-GIC) (Fuji II LC encapsulated, Photac-Fil and Vitremer). The fluoride release of matured restorative GIC was measured as a function of time, after four repeated fluoridations in a 2% NaF aqueous solution for 1 h. This release was corrected for the intrinsic release as determined with a control group. It was demonstrated that application of fluoride is capable of recharging GIC but the subsequent high fluoride release only lasts for one or a few days. Moreover, the fluoride release behaviour depends on the cement formulation. Comparable to the intrinsic release, the net fluoride release after fluoridation is composed of a short- and a long-term process, the former being predominant after fluoridation. The total amount of fluoride released according to the short-term process increases with consecutive fluoridations. This is especially pronounced for the RM-GIC, who exhibit a relatively slow release after fluoridation as compared to the conventional GIC. An explanation for these results is suggested on the basis of the physicochemistry of the setting reaction of the cements and of the fluoridation process. PMID- 10674813 TI - Osseous tissue reaction around hydroxyapatite block implanted into proximal metaphysis of tibia of rat with collagen-induced arthritis. AB - This study was conducted to investigate the influence of osteoporosis on new bone formation around a hydroxyapatite (HA) block implanted into the proximal metaphysis of the tibia of rats with collagen-induced arthritis (CIA). Ten rats were immunized with an emulsion of bovine type II collagen and Freund's complete adjuvant (arthritis group). Another 10 rats, which were not immunized were used as the control group. Seventeen days after immunization, HA block was implanted into the proximal metaphysis of the tibia. Four weeks after implantation, all rats were killed. The serum level of tetrate-resistant acid phosphatase (TRAP), bone mineral density (BMD) in the proximal metaphysis of the tibia and the affinity index in the arthritis group were 28.0+/-3.5 IU/ml, 130.3+/-28.7 mg/cm2 and 77.6+/-10.8%, respectively, and those in the control group were 24.6+/-5.5 IU/ml, 175.9+/-30.5 mg/cm2 and 56.3+/-14.8%. The serum level of TRAP was higher (P < 0.05) and BMD was lower (P < 0.005) in the arthritis group. The amount of new bone formation around the HA block was larger (affinity index, P < 0.05) in the arthritis group than in the control group. These findings suggest that bone formation around HA block might be enhanced even in conditions associated with highly activated bone resorption and bone formation, such as arthritis. PMID- 10674814 TI - Accelerated tissue regeneration through incorporation of basic fibroblast growth factor-impregnated gelatin microspheres into artificial dermis. AB - The objective of this study was to evaluate the effect of incorporation of basic fibroblast growth factor (bFGF)-impregnated gelatin microspheres into an artificial dermis on the regeneration of dermis-like tissues. When used in the free form in vivo, bFGF cannot induce sufficient wound healing activity, because of its short half-life. Therefore, sustained release of bFGF was achieved by impregnation into biodegradable gelatin microspheres. A radioisotope study revealed that incorporation of bFGF-impregnated gelatin microspheres significantly prolonged in vivo retention of bFGF in the artificial dermis. Artificial dermis with incorporated bFGF-impregnated gelatin microspheres or bFGF in solution was implanted into full-thickness skin defects on the back of guinea pigs (1.5 cm x 1.5 cm) (n = 4). Incorporation of bFGF into the artificial dermis accelerated fibroblast proliferation and capillary formation in a dose-dependent manner. However, the accelerated effects were more significant with the incorporation of bFGF-impregnated gelatin microspheres than with free bFGF at doses of 50 microg or higher. We conclude that the gelatin microsphere is a promising tool to accelerate bFGF-induced tissue regeneration in artificial dermis. PMID- 10674815 TI - A long-term study of implanted artificial hydroxyapatite particles surrounding the carotid artery in adult dogs. AB - In this long-term study, we implanted HAP into adult dogs using a silicone chamber attached to the carotid artery to clarify tissue reaction to HAP implantation over a long period. We designed chambers and both hemispheres of the chambers were filled with HAP particles, and were placed around both carotid arteries of seven adult dogs. The implants were removed after 150, 300, 380 days, and histological and ultrastructual examination was undertaken. We observed bone like tissue which was formed where HAP particles were implanted. Immunohistochemical findings showed that osteocalcin and osteonectin were as positive in the bone-like tissue as in normal bone. This study suggests that biological factors from the arterial wall might play an important role in new bone-like tissue forming, and that HAP has a strong osteoconductive ability even at heterogeneous sites. PMID- 10674816 TI - PEG-variant biomaterials as selectively adhesive protein templates: model surfaces for controlled cell adhesion and migration. AB - Our study focused on the role of poly(ethylene glycol) (PEG) in actively regulating the biological responsiveness of protein-adsorbed biomaterials. To this end, we designed PEG-variant biomaterials from a family of tyrosine/PEG derived polycarbonates to present surfaces ranging from low to intermediate levels of PEG concentration, below the PEG level requisite for complete abolition of protein adsorption. We analyzed the effect of PEG concentration on the amount, conformation and bioactivity of an adsorbed model protein, fibronectin, and on the attachment, adhesion strength and motility of L929 fibroblasts. Our results demonstrate that low levels of PEG can regulate not only the extent but also the conformation and specific bioactivity of adsorbed fibronectin. As the PEG concentration was increased from 0 to 6 mol%, the amount of adsorbed fibronectin decreased linearly yet the fibronectin conformation was altered such that the overall bioactivity of adsorbed fibronectin was uncompromised. We report that the degree of cell attachment varied with PEG concentration in a manner similar to the dependence of fibronectin bioactivity on PEG. In contrast, the nature of cell adhesion strength dependence on PEG paralleled the pattern observed for fibronectin surface concentration. Our studies also indicated that the rate of cell migration was inversely correlated with PEG concentration over a narrow range of PEG concentration. Overall, these results highlight the striking ability of PEG-variant biomaterials to systematically regulate the behavior of adsorbed cell adhesion proteins and, consequently, effect cell functions. PMID- 10674817 TI - Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis IV: study of c-myc, c-fos and p53 mRNA expression. AB - In order to investigate how cells recognize biomaterials, mRNA that was expressed in attached human fibroblasts on various substrates was evaluated. The expressed oncogenes (c-fos and c-myc) and tumor suppressor gene (p53) mRNA were then isolated and detected using the RT-PCR method. As a result, c-fos and c-myc mRNA expression varied with respect to differences in the hydrophilicity hydrophobicity of the substrates. Both c-fos and c-myc mRNA expression were low in the fibroblasts that had adhered to hydrophilic surfaces. The tendency of c fos mRNA expression was similar to the adhesion curve of the cells. c-myc mRNA was largely induced in fibroblasts that had adhered to hydrophobic surfaces. p53 mRNA were largely induced in fibroblasts that had adhered to hydrophilic surfaces, while in the cells that had adhered to hydrophobic surfaces, p53 mRNA expression was low. We concluded that the expression of oncogenes and p53 mRNA is a powerful method for studying cell-polymer interactions or the evaluation of the carcinogenic activity of biomaterials. PMID- 10674818 TI - Microdomain structure in polylactide-block-poly(ethylene oxide) copolymer films. AB - Structured surface is an important property of polymer biomaterials for tissue engineering, for its capacity to expose domains with different surface energy and functional groups. For this purpose, amphiphilic A-B-A block copolymers with polylactide (PLA) as A blocks and poly(ethylene oxide) (PEO 3, Mn = 3090; PEO6, Mn = 6110) as B block were synthesized by ring-opening polymerization of either L lactide (L-LA) or DL-lactide (DL-LA), using poly(ethylene glycol)s as macroinitiators and tin(II) octanoate (Sn(Oct)2) as a catalyst. Differential scanning calorimetry (DSC) and electron microscopy were used to study the phase separation of the hydrophobic (PLA) and hydrophilic (PEO) segments in films made of the copolymers and their blends with high-molecular-weight PLA homopolymers. Hydrophilic (PEO) and hydrophobic (PLA) domains were formed at the polymer film surface due to the separation of phases. The phase separation was affected by the copolymer composition and the stereoregularity of PLA blocks in the copolymers. PMID- 10674819 TI - Autocrine nerve growth factor in human keratinocytes. AB - Biologically active nerve growth factor (NGF) is synthesised and released by proliferating normal human keratinocytes. NGF up-regulates the expression of NGF mRNA in keratinocytes. Keratinocytes express both the low (p75)- and the high affinity (TrkA) NGF-receptors, which are located in the basal layer of the epidermis. K252, a specific inhibitor of trk phosphorylation, blocks NGF-induced keratinocyte proliferation, in absence of exogenous NGF. Normal keratinocytes over-expressing TrkA proliferate better than control transfectants, while the NGF mimicking anti-Trk antibody induces an increased keratinocyte proliferation in Trk over-expressing cells as compared to mock transfected keratinocytes. In addition, NGF over-expressing keratinocytes proliferate better than mock transfected cells. K252, by blocking TrkA phosphorylation, induces apoptosis in normal keratinocytes, but not in keratinocytes over-expressing bcl-2. Furthermore, NGF transfected keratinocytes are protected from UV-B-induced keratinocyte apoptosis, by maintaining constant levels of Bcl-2 and Bcl-xL . Taken together these results support the concept of an autocrine survival system sustained by NGF and its high-affinity receptor in human keratinocytes. Because NGF and Trk levels are highly expressed in psoriasis. one could speculate that NGF autocrine system plays a role in the mechanisms associated with this and other hyperproliferative skin conditions, including cancer. PMID- 10674820 TI - Influence of aging and cell senescence on telomerase activity in keratinocytes. AB - Telomeres, which exist in eukaryotic chromosome ends in specialized G-rich TTAGGG structure, protect the ends from degradation or fusion. On the other hand, telomerase is a ribonucleoprotein complex enzyme that synthesizes TTAGGG repeat sequences at the ends of eukaryotic chromosomes. Previous studies suggested that telomere length and telomerase activity cooperate in aging and immortalization of cells. Here, we examined telomere length and telomerase activity in keratinocytes from seven human subjects, including a patient with Werner's syndrome. Telomere length in keratinocytes from healthy individuals was shortened with aging. However, telomerase activity from an individual aged 42 years was reduced, compared with that from a 0 year old individual. Passages of keratinocytes reduced telomerase activity significantly in F2 and F3 keratinocytes from 0 and 42 year old individuals. Withdrawal of either EGF or amphiregulin from medium resulted in down-regulation of telomerase activity. These results suggest that telomere length and telomerase activity in primary cultured keratinocytes may be one of the parameters for cell senescence. However, there remain obscure factors such as ultraviolet-B radiation and growth factors. PMID- 10674821 TI - Exclusion of linkage between dyschromatosis symmetrica hereditaria and chromosome 9. AB - Dyschromatosis symmetrica hereditaria (DSH) is an autosomal dominant pigmentary disorder, first reported by Toyama in 1910. It is characterized by a mixture of hypopigmented and hyperpigmented macules of various sizes on the backs of the hands and feet. The disease gene of DSH and its chromosomal localization have not yet been identified. A family with DSH and idiopathic torsion dystonia (ITD), a rare neurological disease, was recently reported. Therefore, we speculated that there was a linkage between the DSH gene and the ITD gene, named DYT1 and localized on chromosome 9, and performed linkage analysis between DSH and microsatellite markers on chromosome 9 in three Japanese DSH families (36 patients in total). We obtained a LOD score of < -2 over the whole region of chromosome 9 encompassing DYT1. Thus, we conclude that there is no linkage between DSH and DYT1 as well as any region of chromosome 9. PMID- 10674822 TI - Fas/Fas ligand-mediated apoptosis of murine Langerhans cells. AB - Epidermal Langerhans cells (LC) are potent antigen-presenting cells (APC), that play a crucial role in initiating cutaneous immune responses. The Fas/Fas ligand pathway has been implicated as an important cellular pathway in the regulation of peripheral immunity. The morphologic, functional and phenotypic characteristics of LC are becoming well-characterized. However, the mechanisms involved in eliminating LC are poorly understood. In this report, we demonstrated that murine epidermal LC constitutively express the Fas antigen (CD95) and the expression was up-regulated by the addition of IFN-gamma in cultures. Interestingly, epidermal LC underwent apoptosis by the addition of both recombinant soluble Fas ligand (FasL) and IFN-gamma, but not by FasL alone. These results suggest that LC may acquire the susceptibility to Fas-mediated apoptosis through up-regulation of the Fas expression by IFN-gamma derived from activated T cells and that the elimination of LC may be important for preventing excess cutaneous inflammatory diseases. PMID- 10674823 TI - Loss of heterozygosity of adenomatous polyposis coli gene in cutaneous tumors as determined by using polymerase chain reaction and paraffin section preparations. AB - It has been suggested that an alteration in the adenomatous polyposis coli (APC) gene, which is a tumor suppressor gene, is one of the earlier events in carcinogenesis of some adenocarcinomas. We undertook this study to determine the prevalence of loss of heterozygosity (LOH) of the APC gene in several kinds of cutaneous tumors. Fifty-seven unrelated Japanese patients were examined for analysis of the APC gene. The 57 cases consisted of extramammary Paget's disease, squamous cell carcinoma (SCC), eccrine poroma and porocarcinoma, metastatic tumor of rectal adenocarcinoma and malignant melanoma. DNA was extracted from the tumor and normal portions dissected from the formalin-fixed paraffin-embedding sections and amplified with the use of the PCR. The amplified DNA was examined for LOH in the APC gene. Seven samples of 32 heterozygous persons of APC gene (three out of seven eccrine poromas, two eccrine porocarcinomas and two metastatic tumors of rectal adenocarcinoma) showed for LOH in the APC gene. None of the heterozygous samples from the extramammary Paget's disease (11), SCC (five) and melanoma (five) showed LOH. These results suggest that tumor or tumor suppressor genes, other than the APC gene, may be responsible for extramammary Paget's disease and SCC and that LOH involving APC may have some relevance to the formation and progression of eccrine tumors as in rectal tumors. PMID- 10674824 TI - Production of tissue inhibitor of metalloproteinase-1 and -2 by cultured keratinocytes. AB - The imbalance between metalloproteinases and their inhibitors in tissue remodelling is likely to play an important role in various pathologic conditions. In order to understand the role of keratinocytes in regulating extracellular matrix degradation in skin, we analyzed the production of metalloproteinase inhibitors in keratinocytes. Tissue inhibitor of metalloproteinase-1 (TIMP-1) and tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA were detected in cultured human keratinocytes and mouse transformed keratinocyte cell line (KCMH-1) cells by RT-PCR. On several column chromatography separation steps of the KCMH-1 conditioned medium, two specific inhibitors for mammalian collagenase were purified showing an Mr of 29 kDa and an Mr of 22 kDa, respectively. The analysis of their N-terminal amino acid sequence revealed that two inhibitors were TIMP-1 and TIMP-2. The final preparation of TIMP-1 had a specific activity of 56000 U/mg and that of TIMP-2 had a specific activity of 26200 U/mg. Our results suggest that keratinocytes take part in tissue remodelling in skin in secreting both TIMP 1 and TIMP-2. PMID- 10674825 TI - Reconstruction of human hard-palate mucosal epithelium on de-epidermized dermis. AB - Artificial hard-palate mucosa equivalents were reconstructed using keratinocytes derived from normal human hard-palate and de-epidermized dermis. Reconstructed hard-palate mucosal epithelium formed in three-dimensional culture was compared to native hard-palate mucosal epithelium and reconstructed oral buccal mucosal epithelium with regard to keratin expression. Artificial hard-palate mucosal epithelium reconstructed in medium with delipidized serum showed a differentiation pattern similar to that of hard-palate epithelium in vivo. The present study also confirmed that keratinocytes derived from hard-palate mucosa are intrinsically different from those of nonkeratinizing oral surfaces. PMID- 10674826 TI - CD40 ligation inhibits trichilemmoma cell proliferation and induces IL-6 production. AB - CD40 is a member of the tumor necrosis factor receptor superfamily expressed by B cells, monocytes, dendritic cells, epithelial cells, and hematopoietic progenitor cells. Recently, CD40 has been reported to also be expressed on human epidermal cells. We have elucidated the function of CD40 on epidermal tumor cells and have found that trichilemmoma (KTL-1) cells constitutively express CD40 and respond to CD40 ligation by anti-CD40 mAb (EA-5) with a significant decrease in proliferation. We were also able to demonstrate that KTL-1 cells respond to CD40 ligation by EA-5 with the up-regulation of interleukin-6 (IL-6) mRNA expression. Together, the results suggest that CD40 on KTL-1 cells may function to regulate their proliferation associated with the induction of IL-6 production. PMID- 10674827 TI - Expression pattern of the bcl-2 homologous protein bad in normal skin, psoriasis vulgaris and keratinocytic tumors. AB - Previous investigations focused on the mechanisms and regulation of apoptotic process have found that bcl-2 and its homologous proteins are central regulators of the mitochondrial phase of apoptosis. Expression of several members of the bcl 2 family has been studied in various tissues including skin under normal as well as disease conditions. In this report, we investigated the expression of bad, the pro-apoptotic member of the BH3 subfamily, in normal and psoriatic epidermis, keratoacanthoma, and basal and squamous cell carcinomas. Normal and psoriatic epidermis showed accentuation of the staining in the lower suprabasal compartment. A weak, predominantly diffuse staining pattern was observed in the upper epidermis of psoriatic plaques. Keratoacanthoma showed strong but diffuse immunostaining for pro-apoptotic bad, however we found only weak bad expression in squamous cell carcinoma. Seven out of 15 basal cell carcinomas failed to express bad protein. There was no correlation between bad positivity and depth of tumour infiltration. Our observation suggests that the pro-apoptotic bad may function as one of regulators involved in the maintenance of epidermal homeostasis and this function could be altered depending on the disease state. PMID- 10674828 TI - Firm stroking of human skin leads to vasodilatation possibly due to the release of substance P. AB - Many animal species invest a large amount of time in grooming behavior without deriving any apparent benefit. In order for this behavior to have survived, however, it must confer some survival advantage. In seven of eight humans tested, an elevation in the skin's temperature was documented after massaging of the cheeks of the face. The elevation of the skin's temperature reached a plateau after about 40 min of massaging and was correlated to visible erythema. This effect could be inhibited by repeated pretreatment of the skin with topical capsaicin, a chemical that results in the release of substance P from peripheral nerve endings. Thus, it appears that the temperature elevation induced by stroking of human skin is controlled, at least in part, by release of the neurotransmitter, substance P. In conclusion, it appears that the release of neurotransmitter(s) may be the survival advantage that grooming confers to animals. PMID- 10674829 TI - Role of amniocentesis for the diagnosis of subclinical intra-amniotic infection in preterm premature rupture of the membranes. AB - The clinical role of amniocentesis in the management of pregnancies that are complicated by preterm premature rupture of the membranes remains unclear. The indiscriminant use of expectant management, corticosteroids, and empiric antibiotic therapy without knowledge of the presence or absence of intra-amniotic infection poses underappreciated risks to the fetus. This clinical opinion presents the argument that amniocentesis should be an integral part of the management of patients with preterm premature rupture of the membranes. The technical aspects of amniocentesis, the associations between subclinical infection and neonatal morbidity, and the limitations of current interventions are reviewed, and suggestions for future studies that are sorely needed are offered. PMID- 10674830 TI - Ethical consideration of maternal participation in clinical research. AB - This review explores some of the implications for a balanced approach to the ethics of maternal participation in clinical research, from the perspective of the fetus as a patient, which is a central concept in obstetric ethics. The review therefore begins with an account of this topic, followed by an examination of some of its ethical implications for a balanced approach to the ethics of clinical research that involves pregnant women. PMID- 10674831 TI - Life style, work and stress, and pregnancy outcome. AB - Several environmental factors affect the fetus and thereby the outcome of pregnancy. Recent studies have confirmed a relation between stress and pregnancy outcome; furthermore they have indicated that biological measures of stress may predict risk of complications. Altered sex ratio may be an interesting way of measuring the effect of stress during pregnancy. Stress and work load during pregnancy seem to be related to time until conception and to becoming pregnant through assisted reproduction. Drinking large amounts of alcohol is hazardous, but drinking one drink per day appears to be safe. The effect of passive smoking continues to be a matter of debate. PMID- 10674832 TI - Advances in management of Type 1 diabetes and pregnancy. AB - Obstetricians will need to update themselves on the recent changes in terminology and in diagnostic criteria for Type 1 diabetes mellitus. This still remains a high-risk obstetric situation, in spite of optimistic reports from centres of excellence. Pregnancy-associated hypertension may be closely related to insulin resistance. A number of concepts and hypotheses are based on the central role of insulin in fetal development. PMID- 10674833 TI - First trimester screening for fetal abnormalities. AB - The debate over the application of nuchal translucency measurement in Down's syndrome screening is still unresolved in some clinicians' minds. Although different authors report a range of sensitivities for Down's syndrome, none question the validity of the association between increased nuchal translucency and fetal aneuploidy. The published literature reveals a lack of congruence over a standard, reproducible method for measuring nuchal translucency. Only with the adoption of uniform methodology, and the establishment of international standards for nuchal translucency measurement, is the true potential of this test likely to be realized. The increasing use of first trimester ultrasound has focused attention on the value of this investigation in confirming fetal viability, estimation of gestational age, and screening for congenital abnormality. This review summarizes the role of ultrasound and maternal serum biochemistry in first trimester screening. PMID- 10674834 TI - Physiologic basis of antenatal testing. AB - Hypoxic injury accounts for approximately 20 to 40% of stillbirths in published autopsy series. Antepartum fetal testing modalities can discriminate between the fetus at risk for hypoxic injury and the normally oxygenated fetus. These modalities have resulted in a significant reduction in this category of stillbirths. The physiologic underpinning for the antepartum testing is reviewed with a brief discussion of the two major approaches to testing. PMID- 10674835 TI - General obstetrics. PMID- 10674836 TI - Diabetic ketoacidosis in type 2 diabetes mellitus. PMID- 10674837 TI - Cardiac involvement of female carrier of Duchenne muscular dystrophy. PMID- 10674838 TI - Takayasu's arteritis and giant cell (temporal or cranial) arteritis. PMID- 10674839 TI - Hepatic manifestations of the antiphospholipid syndrome. PMID- 10674840 TI - HLA-B39 and asymmetric arthritis. PMID- 10674841 TI - The occurrence of diabetic ketoacidosis in adults. AB - OBJECTIVE: To analyze the occurrence of DKA in Chinese adults. METHODS AND PATIENTS: We retrospectively reviewed the medical records of adults presenting with DKA in a tertiary referral center from January 1992 to December 1997. We classified these patients into 3 groups: type 1, type 2 and new-onset diabetes. Clinical features and follow-up treatment were analyzed. RESULTS: One hundred and twenty patients with 141 episodes of DKA were included; 77 episodes (54.6%) were classified as being caused by type 2, 32 (22.7%) by type 1 and 32 (22.7%) by new onset DM. The average age of type 2 patients was significantly higher. Of the 25 new-onset patients with follow-up for at least 12 months, 11 were not taking insulin. Of these 11 patients, 6 had a family history of DM and 5 had BMI greater than 26.4 kg/m2. The fasting plasma C-peptide values at various times of follow up varied from 2.3 to 9.5 ng/ml in 6 of the 11 DKA-onset patients. CONCLUSION: In type 2 patients, the occurrence of DKA is usually associated with old age and another severe illness. "DKA-onset type 2 DM" reported in African-Americans and in Japanese is also observed in Chinese. PMID- 10674842 TI - Cardiorespiratory responses during cycle ergometer exercise with different ramp slope increments in patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: The ramp exercise test has been widely used to evaluate cardiopulmonary responses to an incremental exercise load. This study was performed to clarify whether different slopes of the ramp exercise test influence exercise tolerance, exercise limiting factors, and respiratory pattern in patients with chronic obstructive pulmonary disease (COPD). SUBJECTS AND METHODS: We applied three different slopes (5 W/min, 10 W/min and 20 W/min) of the ramp exercise test in 9 patients with COPD and evaluated cardiopulmonary responses. RESULTS: There were no significant differences in peak oxygen uptake, anaerobic threshold (AT), minute ventilation, heart rate, arterial oxygen saturation, expired tidal volume, or respiratory rate at the maximal load among the three different ramp exercises tested. AT could be determined in six of nine patients (67%) at the slope of 5 W/min, in 8/9 (89%) at the slope of 10 W/min, and in 9/9 (100%) at the slope of 20 W/min. CONCLUSION: The findings suggest that the ramp slope does not affect exercise tolerance, exercise limiting factors, or respiratory patterns and each of these ramp slopes is useful for the evaluation of COPD. Ramp slopes of 10 W/ min or 20 W/min should be appropriate for the determination of AT. PMID- 10674843 TI - Chromogranin A expression in hepatocellular carcinoma in a patient with germline MEN1 gene mutation. AB - Hepatocellular carcinoma (HCC) was found in a patient with multiple endocrine neoplasia type 1 (MEN 1). The intriguing finding was that the HCC in the patient was positively stained for chromogranin A (CgA), a cellular marker for endocrine and neuroendocrine tumors. The patient had a pancreas endocrine tumor and type C hepatitis, that made pathological diagnosis of the origin of the tumor complicated. PMID- 10674844 TI - Endoscopic resection of small inflammatory fibroid polyp of the colon. AB - Inflammatory fibroid polyp (IFP) is a solitary intestinal lesion of unknown etiology. Although IFP is benign, laparotomy for the resection of colonic IFP is performed in most cases because the polyp is usually large. We report a successful endoscopic resection of cecal IFP. It is considered that colonic IFP should be resected endoscopically if the polyp is small and is located submucosally. PMID- 10674845 TI - Right ventricular cardiomyopathy showing right bundle branch block and right precordial ST segment elevation. AB - A 73-year-old man who had a family history of sudden death, experienced syncope. His electrocardiogram (ECG) presented right bundle branch block and right precordial ST segment elevation which are findings identical with those in Brugada syndrome. The cardiac MRI showed right ventricular mild dilatation, and endomyocardial biopsy revealed fatty replacement of myocardial fibers. Though no ventricular tachyarrhythmias were induced during an electrophysiologic test, the effects on ECG of antiarrhythmic agents and autonomic modulations were similar to those in Brugada syndrome. This case may suggest the relationship between Brugada syndrome and right ventricular cardiomyopathy. PMID- 10674846 TI - A female carrier of Duchenne muscular dystrophy complicated with cardiomyopathy. AB - A 45-year-old female carrier of Duchenne muscular dystrophy (DMD) complicated with cardiomyopathy is described. She had no symptoms of muscle weakness or heart failure. Her chest X-ray film revealed marked cardiomegaly. Echocardiogram showed marked enlargement and severe hypokinesis of the left ventricle. In myocardial scintigraphic images, perfusion defects of the myocardium were revealed. Dystrophin immunostaining of myocardial biopsy specimens showed a mosaic pattern of dystrophin-negative and -positive fibers. Cardiomyopathy is sometimes the only clinical symptom in female carriers of DMD. They are thought to be in a high risk group for developing heart failure. PMID- 10674847 TI - Successful catheter ablation against ventricular tachycardia associated with myotonic dystrophy. AB - Myotonic dystrophy (MD) is characterized by myotonia and muscular dystrophy and cardiac involvement with tachy-arrhythmia is rarely encountered. We report a case of MD complicated with severe left ventricular hypofunction and incessant ventricular tachycardia (VT) with varying heart rates. The morphology of VT suggested that it originated from the right ventricular outflow tract, and electrophysiological study disclosed that the mechanism of VT was abnormal automaticity. Catheter ablation was performed to treat this VT. The patient had a cardiomyopathy with normal coronary arteries. The specimen of RV biopsy showed moderate hypertrophy, mild fat infiltration and slight fibrosis. These findings are histologically consistent with myotonic dystrophy. PMID- 10674848 TI - Acute intermittent porphyria associated with transient elevation of transaminases during an acute attack. AB - A 44-year-old woman with acute intermittent porphyria (AIP) was admitted to Kudanzaka Hospital because of abdominal pain. A cholecystectomy was performed in another hospital without improvement. On admission, her transaminases were elevated to greater than 1,000 mU/ml. After an intravenous drip of mainly glucose, her transaminases returned to normal. Her acute attacks occurred during stress, and she died of respiratory failure after repetitive acute episodes. AIP should be included in a list of the differential diagnosis of gastrointestinal diseases, neurosis, and hysteria. This is the first case of AIP accompanied by transient marked elevation of transaminases during an acute attack. PMID- 10674849 TI - Familial dysalbuminemic hypertriiodothyroninemia in a Japanese kindred. AB - A 56-year-old Japanese housewife had been diagnosed as having Graves' disease and was treated with methimazole. When she was referred to our hospital, the serum T3 level was high irrespective of high TSH level. High serum T3 levels were also observed in two out of her three sisters. Electrophoresis revealed that binding of 125I-T3 to serum albumin was markedly increased whereas the binding of 125I-T4 to serum albumin was slightly increased in the three sisters whose serum T3 levels were high. These data indicate that the presence of an albumin variant is the cause of hypertriiodothyroninemia in this family. PMID- 10674850 TI - Electrolyte disorders following massive insulin overdose in a patient with type 2 diabetes. AB - We present a case of a 47-year-old man with Type 2 diabetes mellitus who attempted suicide with 2,100 U of insulin injected subcutaneously. Administration of dextrose intravenously was required to maintain the blood glucose concentration normally for 5 days. Moreover, hypokalemia, hypophosphatemia, and hypomagnesemia were also seen for 24 hours after insulin injection. The serum phosphorus and magnesium concentrations decreased to nadirs of 1.6 mg/dl and 1.6 mg/dl respectively 7 hours after insulin injection. Electrolyte disorders other than hypokalemia may be induced in hypoglycemic patients by massive insulin overdose. PMID- 10674851 TI - Results of surgery for a compound adrenal tumor consisting of pheochromocytoma and ganglioneuroblastoma in an adult: 5-year follow-up. AB - A rare, compound adrenal tumor consisting of ganglioneuroblastoma and pheochromocytoma was completely resected in an adult woman. Most of the tumor was occupied by the ganglioneuroblastoma component. This ganglioneuroblastoma was an intermixed tumor, which is known to have a favorable prognosis in children. Based on the lack of spread, the resectability of the tumor, and the histology of the ganglioneuroblastoma, no adjuvant therapy was employed. There was no evidence of recurrence at the 5-year follow-up. This suggests that adjuvant therapy may not be necessary in these compound tumors. PMID- 10674852 TI - Recurrent hemoptysis in tuberculosis with non-cavitary lung disease as a symptom of mild hemophilia A in a young adult. AB - Hemoptysis in patients with tuberculosis is usually associated with smear positive and cavitary lung disease. The present case describes a patient suffering from recurrent hemoptysis associated with tuberculosis who had smear negative and non-cavitary lung disease, and who was subsequently diagnosed as having mild hemophilia A. Although mild hemophilia A sometimes escapes detection until adolescence, there has been no reported case of mild hemophilia A detected by recurrent hemoptysis due to pulmonary tuberculosis. Here, we report a rare case of recurrent hemoptysis in a patient with tuberculosis who had smear negative and non-cavitary lung disease and who was finally shown to have hemophilia A. PMID- 10674853 TI - Acute eosinophilic pneumonia caused by cigarette smoking. AB - It has been suggested that acute eosinophilic pneumonia (AEP) is associated with cigarette smoking because in Japan, the patients with AEP are young and have a high incidence of short-term smoking history. However, there has been no direct evidence to support that cigarette smoke causes AEP. Herein is reported the first case showing the direct evidence and a long-term clinical course of cigarette smoking-induced AEP, in which tolerance to repeated resumption of smoking cigarettes might have occurred. We should pay attention to the history of cigarette smoking in seeing patients with AEP, especially in young patients. PMID- 10674854 TI - Takayasu's arteritis in a 69 year-old woman. AB - Takayasu's arteritis and temporal arteritis share many clinical and pathological features. The most discriminatory feature between the two diseases is the age at onset; the mean age at onset of the disease was reported as being 26 years for Takayasu's arteritis and 69 years for temporal arteritis. Here we report a 69 year-old woman who presented with a weak right radial artery pulse. The ethnic background and the presence of vascular insufficiency of the right upper extremity and the absence of clinical signs such as shoulder stiffness and tender scalp indicate that her diagnosis is Takayasu's arteritis. It must be emphasized that the two conditions could be differentiated based on the clinical findings even in a patient as old as 69 years old. PMID- 10674855 TI - Sjogren's syndrome complicated with autoimmune hepatitis and antiphospholipid antibody syndrome. AB - A 56-year-old Japanese female simultaneously developed thrombocytopenia, sicca symptoms, and an elevation of transaminase. Antiphospholipid antibodies were detected in her serum. The presence of anti-SS-A antibodies in the serum and sialectasis, disclosed by sialography, suggested the presence of primary Sjogren's syndrome (SjS). The laboratory data and the biopsy of the liver showed compatible findings with autoimmune hepatitis (AIH). Thrombocytopenia and liver dysfunction satisfactorily responded to corticosteroid. To our knowledge, this is the first reported case of SjS with AIH and antiphospholipid antibody syndrome (APAS). Analysis of serum cytokine levels showed a predominance of Th0-Th1 response, which is not compatible with AIH, in this complicated autoimmune state. PMID- 10674857 TI - Bilateral facial palsy following trigeminal zoster with zoster oticus. PMID- 10674856 TI - A family with cases of adult onset Still's disease and psoriatic arthritis. AB - Adult onset Still's disease is recognized as an adult variant of the systemic form of juvenile rheumatoid arthritis, whose disease-predisposition is still debated. On the other hand, the association between HLA subtypes and several groups of seronegative arthritis including psoriatic arthritis has been well documented. This report describes a family where adult onset Still's disease in a young man and psoriatic arthritis in his father were seen. Both patients were HLA B39-positive, which was likely playing important pathogenic roles in the latter case. Clinical and immunological aspects of HLA-B39-related inflammatory diseases are also discussed. PMID- 10674858 TI - Overuse of granulocyte colony-stimulating factor in acute myeloid leukemia, aplastic anemia and myelodysplastic syndrome. PMID- 10674859 TI - Patent foramen ovale and "catastrophic" antiphospholipid syndrome. PMID- 10674860 TI - Does amblyopia protect against age-related maculopathy? PMID- 10674861 TI - Pseudoexfoliation syndrome in a patient with lattice corneal dystrophy. AB - We report the case of a 70-year-old female who presents lattice corneal dystrophy type I in association with pseudoexfoliation syndrome. This association has never been reported in patients not affected by systemic amyloidosis. PMID- 10674862 TI - The incidence of nontraumatic phakic rhegmatogenous retinal detachment in Split Dalmatia County, Croatia. AB - We studied the incidence of nontraumatic phakic rhegmatogenous retinal detachment (RRD) in a defined population of Split-Dalmatia County, Croatia. In this clinic based study, 272 of a population of 465,947 developed RRD during an 11-year period (1988-1998). The annual incidence was, therefore, 5.3 per 100,000 population. The highest risk for RRD was between the ages of 60 and 69 years. PMID- 10674863 TI - Ligneous conjunctivitis: a clinicopathologic study of 3 cases. AB - The clinical, histopathologic features, and treatment outcomes in 3 patients with ligneous conjunctivitis are described. Bilateral, idiopathic membranes occurred in the palpebral conjunctiva in 2 patients. In 1 patient, unilateral conjunctival changes occurred in the bulbar conjunctiva, at the site of pterygium excision. Treatment included topical hyaluronidase, chymotrypsin, heparin, and cyclosporine and surgical excision with limited or no success. In one patient, conjunctival autografting from the normal fellow eye resulted in pseudomembrane formation at the donor site in the previously unaffected eye. Histopathological evaluation of excised membranes revealed the presence of amorphous eosinophilic hyaline material and chronic inflammatory cells. Immunohistochemical study revealed a predominance of T-lymphocytes. This case series confirms the recalcitrant clinical course of ligneous conjunctivitis. Conventional treatment modalities described in literature were not useful in the management of this condition. Surgical manipulation of the unaffected fellow eye in patients with unilateral disease can result in pathologic conjunctival changes, and is best avoided. PMID- 10674864 TI - A case of episcleral neurofibroma. AB - PURPOSE: To report a rare case of episcleral neurofibroma and discuss the possible differential diagnoses. METHODS: Case report of a 36-year-old man who presented with a painless epibulbar mass of the left eye. We describe the clinical and histopathologic features of the tumour and compare it with other tumours which may have a similar clinical presentation. RESULTS: An excisional biopsy of the tumour was performed. Histopathologic examination revealed the tumour to be an isolated episcleral neurofibroma. CONCLUSION: It is often difficult to clinically differentiate this tumour from other conditions. Because of the slow growth of neurofibromas and its slow risk of malignant transformation, these lesions may be observed periodically for progression. Surgical excision may be performed if the lesion is found to be progressively enlarging in size. PMID- 10674865 TI - Trabeculectomy: a retrospective follow-up of 700 eyes. AB - A major focus of our study was to determine the value of postoperative intraocular pressure (IOP) in predicting the outcome of trabeculectomy (TE). The medical charts of 547 patients undergoing glaucoma filtering surgery at the Department of Ophthalmology of the University of Cologne from 1987 to 1996 were reviewed. The status of the visual field, level of visual acuity, appearance of the bleb, cup/disc ratio and IOP were studied. Pre- and post-operative glaucoma medication was recorded. The eyes with congenital glaucoma and those treated with antimetabolites were excluded. The results are presented with particular emphasis being placed not only on intraocular pressure (IOP) control but also on the progression of glaucomatous damage (deterioration of visual field or disc damage) and the decrease of visual acuity. The tonometric success rate of TE in controlling the IOP < 21 mmHg was 61%. Defining the rigid criteria for success of trabeculectomy as an IOP < 21 mmHg, no further visual field loss, no disc damage and no additionally required surgical intervention due to glaucoma, the success rate decreased to 44%. The results indicate that other factors than normalization of IOP determine the success rate of TE. Should trabeculectomy be the therapy of first choice in the early stage of glaucoma? Should trabeculectomy fail to control the IOP in the first eye, would this allow options, such as the use of antimetabolites in the second eye? PMID- 10674866 TI - Why study vascular factors in glaucoma? AB - There is clinical and experimental evidence that both increased intraocular pressure and disturbed circulation are involved in the pathogenesis of glaucomatous damage. Among the many factors discussed, decreased blood pressure and vasospasm are the most important, and these factors may, at least in part, be therapeutically influenced. The basic underlying disorder might be a vascular dysfunction leading to local vasospasm and to systemic hypotension. PMID- 10674867 TI - Cataract surgery: an analysis of patient satisfaction with medical care. AB - INTRODUCTION: Patient satisfaction is a good performance indicator for measuring the quality of health care delivered by hospitals. Satisfaction is understood to be the positive difference between users' perceptions of their experience at the moment of discharge and their expectations at the moment of admission. OBJECTIVE: To evaluate the expectations and the perceived quality of care in cataract surgery patients attending Hospital Clinic (HC). METHOD: A two-stage descriptive study carried out at the HC in Barcelona, Spain. The target population consisted of patients operated on for cataracts during the 1996 calendar year. Two study groups were established: Group I, patients attending outpatient service before admission; and Group II, patients attending outpatient service after surgery. After informed consent was obtained, the patients were directly interviewed by three researchers especially trained for that purpose. The questionnaire included demographic variables and 31 questions related to expectations, all to be answered by means of a 7 points Likert's visual scale. All statistical calculations were performed using the SPSS program for Microsoft Windows. RESULTS: A total of 148 interviews were performed: 80 (54.1%) in Group I and 68 (45.9%) in Group II. The mean age was 64.2 +/- 11.6 years. The difference between scores at admission and at discharge was nearly significant (p = 0.064) for the "information" component. CONCLUSIONS: The study of several fields where the patient's expectations are higher or lower contributes to prioritizing efforts to improve quality. There is a need for a frequent review and update of any patient satisfaction evaluation tools that are used. PMID- 10674868 TI - Uveoscleral outflow in dog's eye: role of several enzymes. AB - The morphological pattern of several enzymes (succinic dehydrogenase--SDH, glucose-6-phosphate dehydrogenase--G6PDH and lactic dehydrogenase--LDH) was evaluated in normal dog eyes. Special attention was paid to the uveo-scleral tissue. Cryostatic sections of dog eye were stained with toluidine blue for the recognition of the microanatomical details or with histoenzymatic methods for SDH, G6PDH and LDH activities using sodium succinate, glucose-6-phosphate and sodium lactate as substrates respectively, nicotinamide adenine dinucleotide (NAD) as a reducing agent and sodium nitro-blue-tetrazolium as a colouring substance. A moderate positive reaction for SDH and a strong positive reaction for LDH were observed in the uveoscleral tissue, while G6PDH gave negative staining. Some considerations regarding a possible active role of these enzymatic activities to the aqueous humor outflow are suggested. PMID- 10674869 TI - Use of perfluorocarbon liquids, silicone oil, and 5-fluorouracil in the management of experimental PVR. AB - PURPOSE: To evaluate the toxicity and efficacy of 5-fluorouracil (5-FU) in combination with perfluoroperhydrophenanthrene (Vitreon), silicone oil, or a combination of silicone oil and Vitreon in a ratio of 3:2 in the management of experimental proliferative vitreoretinopathy (PVR). METHODS: Toxicity study. Seventy rabbit eyes underwent vitrectomy followed by intravitreal injection of 5 FU in doses of 800, 400, or 200 microg: Group 1, 5-FU alone; Group 2, 5-FU plus 1 mL Vitreon; Group 3, 5-FU plus 1 mL silicone oil; Group 4, 5-FU plus 0.6 mL silicone oil and 0.4 mL Vitreon; Group 5, 0.6 mL silicone oil plus 0.4 mL Vitreon. Electroretinography was performed preoperatively and 8 weeks postoperatively before the animals were sacrificed. Efficacy study. Seventy-two rabbit eyes underwent vitrectomy and were injected intravitreally with 100,000 200,000 retinal pigment epithelial cells to induce PVR. Groups were injected with 200 microg 5-FU alone or with 1 mL silicone oil, 1 mL Vitreon, or a combination of 0.6 mL silicone oil and 0.4 mL Vitreon. Others were given only 1 mL Vitreon or 1 mL silicone oil. The animals were followed for 8-12 weeks; PVR was graded using Fastenberg's system. RESULTS: Toxicity study. Eyes given 200 microg 5-FU, silicone, and Vitreon showed mild inflammation and vitritis which resolved in 1 week; the dose was nontoxic by electroretinography and histopathology. Doses of 400 and 800 microg 5-FU were toxic. Efficacy study. Clinical severity of PVR was less in the groups which received 5-FU plus vitreous substitutes when compared to the control groups at all time points. The lowest incidences were in groups given 5-FU plus Vitreon or 5-FU plus Vitreon and silicone oil: 33.33% and 11.11%, respectively. CONCLUSIONS: A dose of 200 microg 5-FU with silicone oil and Vitreon combined was nontoxic to the rabbit retina. The combination of 5-FU, Vitreon, and silicone oil showed significant efficacy in the prevention of experimental PVR. PMID- 10674870 TI - Are ophthalmic surgeons aware that starch powdered surgical gloves are a risk factor in ocular surgery? AB - PURPOSE: To assess the level of awareness among UK ophthalmic surgeons of the potential risks from starch powdered surgical gloves during ophthalmic surgery and to show by electron microscopy that starch granule contamination can occur during ophthalmic surgery. SETTING: A sample (N = 46) of UK ophthalmologists from the North of England, UK. METHODS: Type of glove usage and awareness of the possible risks from starch powdered surgical gloves were assessed by means of a questionnaire sent to ophthalmic surgeons in the North of England. The surface of a polymethylmethacrylate (PMMA) intraocular lens (IOL) handled with a starch powdered surgical glove was examined by electron microscopy for evidence of starch contamination. RESULTS: Of the sampled ophthalmic surgeons (46), 89.1% considered it important to use starch free surgical gloves and the 84.8% already did so. Starch granule contamination was seen by electron microscopy on the surface of a PMMA IOL which had been handled with starch powdered surgical gloves. CONCLUSIONS: Although there has been sporadic attention in the ophthalmic literature to the risks associated with starch powdered surgical gloves in ophthalmology, up to 15% of UK ophthalmic surgeons may still be using starch powdered gloves. The authors show that starch powder contamination of ophthalmic materials can actually occur and remind ophthalmologists that this has been reported in the literature as a possible cause of sterile intra and extraocular inflammation. PMID- 10674871 TI - Early results of punch trabeculectomy. AB - PURPOSE: To evaluate the effectiveness of punch trabeculectomy in controlling intraocular pressure (IOP) in patients with uncontrolled chronic open-angle glaucoma (COAG) undergoing medical treatment. METHODS: This prospective study included 22 patients (27 eyes) with uncontrolled COAC on medical treatment undergoing punch trabeculectomy through a scleral tunnel. The tunnel was created with a 3.5 mm keratome, the tunnel mouth was not sutured but the conjunctival flap was sutured by 8/0 vicryl. The mean surgical time from opening the conjunctiva to closing the conjunctiva was 8 m 59 s. IOP and other parameters were checked on day 1 and subsequently at intervals appropriate to the individual. RESULTS: After a mean period of 22.2 months follow-up the mean IOP was 13.3 mmHg (SD 5.2). There were few transient early post-operative complications; two eyes had a shallow anterior chamber (AC) due to excess drainage, five had IOP above 21 mmHg, which responded to ocular massage, ten eyes had hyphaema less than 1.5 mm and two eyes had no filtration bleb. CONCLUSIONS: Punch trabeculectomy is a rapid and effective method of controlling IOP. The complication rate is moderate and mainly transient. PMID- 10674872 TI - The pure antiestrogen ICI 182780 is more effective in the induction of apoptosis and down regulation of BCL-2 than tamoxifen in MCF-7 cells. AB - There is increasing evidence that induction of apoptosis by antihormones is an important mechanism in regard to their growth inhibitory action on hormone dependent tumors. In this report we have compared the efficiency of tamoxifen (Tam) and the pure antiestrogen ICI 182780 (ZM) to induce apoptosis in the estrogen dependent breast cancer cell line MCF-7. Clear evidence for induction of apoptosis could be demonstrated after treatment with both antiestrogens. Application of the pure antiestrogen ZM led to a significantly higher induction of apoptosis compared to the partial agonistic compound Tam. The ability of the two compounds to induce apoptosis correlated with their growth inhibitory action. On the molecular level administration of ZM led to a time dependent steady decrease of BCL-2 mRNA and protein. Administration of Tam also initially decreased the expression of BCL-2. In contrast to ZM treatment, BCL-2 expression increased again after 8 h of incubation with Tam. After 96 h Tam treated cells expressed BCL-2 levels nearly as high as untreated cells. In general, ZM decreased BCL-2 levels more effectively than Tam. Our results demonstrate that ZM and Tam possess quantitative and qualitative differences in their ability to down regulate BCL-2 expression. The higher ability of the pure antiestrogen to down regulate BCL-2 expression may explain the superiority of the pure antiestrogen to induce apoptosis and to inhibit the growth of MCF-7 cells. PMID- 10674873 TI - Mdm2 sensitizes MCF7 breast cancer cells to cisplatin or carboplatin. AB - Overexpression of Mdm2 in cancer cells with otherwise wild-type p53 is believed to be an alternative mechanism for p53 inactivation during carcinogenesis. Because a number of genetic alterations that inactivate p53, including mutation, homozygous deletion, or viral oncoprotein expression (e.g. HPV16-E6), inhibit DNA repair, we tested the hypothesis that Mdm2 would likewise inhibit DNA repair. Repair of cisplatin-induced DNA damage was reduced in MCF7 cells overexpressing Mdm2, compared to MCF7 cells in which wild-type p53 function was intact. MCF7 Mdm2 cells exhibited preferential sensitivity to cisplatin and carboplatin. MCF7 Mdm2 cells showed a pronounced S-phase arrest after cisplatin treatment, similar to that observed in mutant-p53 cells in the present and prior studies. MCF7 cells with intact wild-type p53, on the other hand, arrested primarily in G2/M phase after cisplatin treatment. These findings indicate that Mdm2 overexpression can recapitulate the effect of p53 mutations on DNA repair of cisplatin lesions. PMID- 10674874 TI - Immunohistochemical expression of internal and external ErbB-2 domains in invasive breast cancer. AB - We tested three ErbB-2 monoclonal antibodies (MoAbs) specific to the intracytoplasmic internal domain (clone CB 11) and the extracellular glycosylated peptide domain (clones CBE1 and Tab250) in 351 primary invasive breast carcinomas. ErbB-2 immunodetection allowed us to differentiate three main groups: group 1 (62.7%) lacked both MoAb ErbB-2 domains (erb -/-); group 2 stained for both domains (erb +/+) (26.5%); group 3 stained for the internal domain only (erb +/-) (10.8%). The relationships among these groups and nodal status (N) were statistically significant, with N+ cases reaching the highest value (89.2%) in the erb +/- group. Lack of immunostaining in the external domain thus seems to be associated with increased metastatic spread. At variance analysis the difference in hormonal receptor content between groups 1 and 3 was not significant; while between groups 1 and 2 it was. The growth fraction of groups 2 and 3 was significantly higher than that of group 1. Our results showed that anti-ErbB-2 MoAb clone CB 11 was able to detect a higher number of ErbB-2 expressing cases than the two that are specific for the external domain (clones Tab 250 and CBE1). Due to the strong association between group 3 cases and the highest metastatic potential, this aggressive group could be identified only with the use of an internal-domain specific MoAb CB 11, which thus seems to present a better discriminative power as a diagnostic marker in the biopathological characterization of breast carcinoma. PMID- 10674875 TI - Survival from contralateral breast cancer. AB - First primary, or unilateral, breast cancer (UBC) cases diagnosed in 1960-89 at the Cancer Institute (WIA), Chennai, India were followed-up until December 31, 1994. Patients with UBC (n = 3163) and those who developed second cancer in the contralateral breast (CBC) after the initial breast cancer (n = 67 or 2.1% of UBC) were analysed. Compared to UBC patients, those who developed CBC were younger at the time of diagnosis of initial breast cancer and had higher frequency of breast cancer among the family members. The relative survival rate takes into account competing causes of death and was estimated as the ratio of observed survival rate to the expected survival rate. The cumulative relative survival from UBC at 5 and 10 years were 51% and 41%, respectively, and the corresponding rates for CBC were 47% and 30%; the survival difference seen between UBC and CBC patients was not statistically significant. The survival rates among younger, middle-aged and older women were significantly different from each other in UBC but not in CBC patients. Both UBC and CBC with early stage disease had a better survival compared to late stage disease. Survival advantage was also seen among both UBC and CBC patients with family history of breast cancer compared to those without. The multivariate analysis by the life table proportional hazards model showed that the age at diagnosis is an independent prognostic factor for breast cancer. The study results should be interpreted in the light of small sample size of second cancers. PMID- 10674876 TI - Bone marrow micrometastases in breast cancer patients. AB - The presence of epithelial cells in bone marrow may be a prognostic factor in breast cancer, and so we evaluated their evolution in treated and untreated patients. A first bone marrow aspirate was obtained from 125 stage I/II breast cancer patients at diagnosis and repeated every 6-8 months; the samples were processed for leukocyte separation, used to prepare cytospin slides, stained with a pool of monoclonal antibodies (MoAb) recognising epithelial antigens, and immunocytochemically processed. The median follow-up was 48 months (range 15-82); 23 patients relapsed, and 14 died. MoAb positive cells were observed in 31.2% of first, 24.3% of second, and 27.8% of third aspirates. In 68/100 pairs of successive aspirates, bone marrow status remained unchanged; in 20 it became negative, and in 12 positive (not statistically significant even after adjusting for adjuvant therapy). An analysis based on Mantel and Byar's approach to time dependent covariates using all 225 aspirates found no statistically significant prognostic difference between the patients with negative and positive bone marrow. Bone marrow status changed over time in about 1/3 of the patients; adjuvant therapy did not affect the probability of its becoming negative or positive. No significant association was found between bone marrow evolution and relapse or death, but the relatively high probability of a change in status over time cannot exclude the possibility that a positive aspirate during the course of breast cancer may be a negative prognostic factor. PMID- 10674877 TI - Prevention of rat mammary carcinoma utilizing leuprolide as an equivalent to oophorectomy. AB - A clinical trial is currently under way to examine the effectiveness of leuprolide as a breast cancer chemopreventive agent and contraceptive. This trial, as well as similar proposed studies, is based on the assumption that leuprolide is as effective as surgical castration in preventing the onset of mammary tumors; however, this has not been well documented in the DMBA animal model. We directly compared leuprolide and oophorectomy in this model and examined a combined therapy of leuprolide/bromocriptine. Twenty-seven day old female Sprague-Dawley rats were randomly allocated into one of eight groups. All rats received a 20-mg dose of DMBA at the age of 55 days. Group 1 (n = 10), no treatment; Group 2 (n = 9), leuprolide (100 microg/kg/day) for eight weeks beginning four weeks prior to DMBA; Group 3 (n = 10), oophorectomy four weeks prior to DMBA with replacement estrogen beginning four weeks following DMBA. Estrogen replacement was achieved with a 0.05-mg estradiol tablet releasing 0.833 microg/day over a 60-day period. Group 4 (n = 10), leuprolide (100 microg/kg/day) initiated two weeks prior to DMBA and continuing for two weeks following DMBA; Group 5 (n = 9), oophorectomy two weeks prior to DMBA with 0.05 mg of estradiol in depot form, releasing 0.833 microg/day, beginning four weeks following DMBA and continuing until week 16 of the study; Group 6 (n = 10), leuprolide (100 microg/kg/day) beginning two weeks prior to DMBA and continuing for the duration of the experiment; Group 7 (n = 10), leuprolide (100 microg/kg/day) for eight weeks beginning two weeks prior to DMBA; Group 8 (n = 9), leuprolide (100 microg/kg/day) and bromocriptine (83 microg/day) for eight weeks beginning two weeks prior to DMBA. At nineteen weeks (15 weeks post DMBA), animals were sacrificed and autopsies performed. One hundred percent of untreated animals developed tumors. No animals undergoing oophorectomy four weeks prior to DMBA or receiving leuprolide four weeks prior to and simultaneously with DMBA developed tumors. In animals pretreated two weeks prior to DMBA with leuprolide or oophorectomy, each group had one animal with tumor development. No tumors developed in the animals receiving ongoing injections of leuprolide. However, one tumor developed in those receiving leuprolide for the first eight weeks beginning two weeks prior to DMBA administration. One animal receiving both leuprolide and bromocriptine developed one tumor. We conclude that chemical oophorectomy (with leuprolide) is as effective as surgical oophorectomy in inhibiting DMBA induced carcinogenesis. PMID- 10674878 TI - Mammaglobin B as a novel marker for detection of breast cancer micrometastases in axillary lymph nodes by reverse transcription-polymerase chain reaction. AB - A novel reverse transcription-polymerase chain reaction (RT-PCR) assay using mammaglobin B gene was developed for detection of breast cancer micrometastases in axillary lymph nodes. Fourteen primary breast cancers and 56 axillary lymph nodes from six patients with primary breast cancer and 15 control lymph nodes from non-cancer bearing patients were subjected to this assay. The transcript of mammaglobin B gene was detected in none of the control lymph nodes, but in all of the 14 primary breast cancers. Eleven out of the 56 lymph nodes from the patients, which were shown to be positive by histological examination, were also proven positive by this assay. On the other hand, fourteen of the 45 (31%) histologically negative lymph nodes were also shown to express mammaglobin B mRNA, which suggested the presence of micrometastases in these lymph nodes. RT PCR using mammaglobin B gene could therefore be a useful tool for detection of micrometastases of breast cancer. PMID- 10674879 TI - Phase III trial of cyclophosphamide, epirubicin, fluorouracil (CEF) versus cyclophosphamide, mitoxantrone, fluorouracil (CNF) in women with metastatic breast cancer. AB - BACKGROUND: The mitoxantrone combination CNF and the epirubicin combination CEF have shown similar activity and less toxicity than the standard CAF combination in metastatic breast cancer (MBC). A prospective randomised study was started to compare safety and activity between CEF and CNF administered using a classical chemotherapeutic schedule in MBC. PATIENTS AND METHODS: From December 1987 to June 1993, 151 patients were randomised to receive cyclophosphamide (C) 100 mg m( 2) p.o. days 1-14, fluorouracil (F) 500 mg m(-2) i.v. days 1 and 8, and epirubicin (E) 30 mg m(-2) i.v. days 1 and 8, or mitoxantrone (N) 6 mg m(-2) i.v. days 1 and 8, every 4 weeks. Seventy-three patients were eligible for CEF and 72 for CNF. RESULTS: Objective responses were observed in 61.6% of the CEF group and 44.4% in CNF group (p = 0.004). The median duration of response was 64 weeks in CEF and 50 weeks in CNF group (p = 0.02) and median time to progression was 51 and 33 weeks, respectively (p = 0.0004). At the time of analysis, all except six patients (one in CNF and five in CEF) had died and the median survival time in the CEF group was longer than in CNF (74.4 weeks vs 51.4 weeks; log-rank chi2 test p = 0.015). CNF produced more hematologic toxicity than CEF (WHO scale; grades 2-4); leucopenia 84% vs 68% (p = 0.03) and thrombocytopenia 17% vs 4.5% (p = 0.01); CEF caused more grade 2 and 3 alopecia: 93% vs 70% (p = 0.001). CONCLUSION: The combination CEF using this schedule and dosage in metastatic breast cancer is more effective with less toxicity than CNF, except for alopecia, and was associated with longer survival. PMID- 10674880 TI - Short and long-term effects on survival in breast cancer patients treated by primary chemotherapy: an updated analysis of a randomized trial. AB - A potential advantage of primary over adjuvant chemotherapy in breast cancer survival had been proposed on theoretical grounds. In 1994, early results of the S6-trial comparing primary chemotherapy vs. adjuvant chemotherapy for operable breast cancer in 390 premenopausal patients had shown significant improvement in survival of the primary chemotherapy arm (p = 0.04). An updated analysis conducted in 1995 showed the disappearance of this difference between the two arms (p = 0.18). In the present analysis, we investigated the potential short and long-term benefits attributable to primary chemotherapy by applying weighted logrank tests designed to assess specifically these effects. Results were compared to those obtained with the classical logrank test. At a median follow-up of 105 months, a significant short-term survival benefit (p = 0.02) in favor of the primary chemotherapy has been shown. However, no long-term survival benefit (p = 0.36) could be documented. The classical logrank test had revealed no significant difference (p = 0.24) between the two groups but the proportional hazard assumption being rejected (p = 0.04), the efficiency of this test can be questioned. Results using the present analysis suggested that primary chemotherapy delayed early death rates, without significantly modifying long-term event rates. It emphasizes that a short-term effect which is not necessarily associated with a long-term benefit may be seen at an early evaluation and disappear later on. PMID- 10674881 TI - Static disease on anastrozole provides similar benefit as objective response in patients with advanced breast cancer. AB - BACKGROUND: This paper reports on the clinical relevance of durable static disease (SD) (> or = 24 weeks) in breast cancer patients treated with the aromatase inhibitor anastrozole. PATIENTS AND METHODS: All patients were part of two prospective, randomised, multicentre studies in postmenopausal women with advanced disease in which megestrol acetate was compared with anastrozole 1 mg. Survival from initiation of treatment was analysed by the response type, i.e., complete response (CR)/partial response (PR), static disease (SD) (> or = 24 weeks), or progressive disease (PD), achieved on therapy. RESULTS: Median survival with anastrozole 1 mg was similar between patients who obtained CR/PR and SD (> or = 24 weeks). Similarly, no difference in survival was observed in patients treated with megestrol acetate who achieved CR/PR and SD. With both treatments patients with CR/PR and SD had improved survival over those patients with PD within 24 weeks. There was no difference between treatment arms for patients showing PD within 24 weeks. CONCLUSIONS: These data confirm that durable SD (> or = 24 weeks) is a clinically useful remission criterion in postmenopausal women with advanced breast cancer with predictive value for overall survival. It also confirms the value of this endpoint with anastrozole, a new generation aromatase inhibitor. PMID- 10674882 TI - Balance of cell proliferation and apoptosis in breast carcinogenesis. AB - We determined the mitotic and apoptotic index through the spectrum of pre invasive ductal breast lesions to invasive carcinoma in search of disturbances in the proliferation/cell death balance in breast carcinogenesis. Seventy-two pure pre-invasive ductal breast lesions (without invasive carcinoma) and 103 invasive breast carcinomas were used. The numbers of mitotic and apoptotic cells were microscopically counted in hematoxylin and eosin stained sections (MI and AI, respectively), and the ratio of the values of MI and AI was calculated for each individual case (M/A index). A distinction was made between well differentiated and poorly differentiated breast lesions, based on histological type and nuclear grade, to arrive at two plausible progression models for breast carcinogenesis. For the well differentiated breast lesions, the MI was rather equal for hyperplasias and well differentiated DCIS, but increased 6-fold from DCIS to well differentiated invasive carcinoma. The AI remained in the same range, resulting in a 4-fold increase of the M/A index. For the poorly differentiated breast lesions, a significant increase in MI and AI was found from hyperplasia to poorly differentiated DCIS. From DCIS to poorly differentiated invasive carcinoma, the MI increased significantly and the AI decreased 2-fold (n.s.), resulting in a 2.5 fold significant increase of the M/A index. In conclusion, the net increase of the number of cells in the transition from well differentiated pre-invasive to well differentiated invasive carcinoma is accompanied by an increase of cell proliferation rather than decrease in apoptosis, suggesting that in these lesions, proliferation related mechanisms are most important in carcinogenesis and progression. In contrast, in poorly differentiated breast lesions, decreased apoptosis seems to be also important in carcinogenesis and progression. At present, we are gathering patients with invasive breast cancer who had a previous biopsy with a pre-invasive lesion to obtain further more direct evidence for this hypothesis. PMID- 10674883 TI - Involvement of nuclear steroid/thyroid/retinoid receptors and of protein kinases in the regulation of growth and of c-erbB and retinoic acid receptor expression in MCF-7 breast cancer cells. AB - Nuclear steroid/thyroid/retinoid receptors and c-erbB membrane receptor tyrosine kinases control epithelial growth and differentiation. Retinoid receptors can dimerize with the vitamin D receptor, the glucocorticoid receptor or the thyroid receptor. Furthermore, multiple c-erbB receptor dimers have been identified. It has been shown that some of these receptor pathways communicate with each other via cross-connected regulatory networks. Molecular interactions between retinoid receptors or estrogen receptors (ER) and c-erbB-2, and between ER and retinoic acid receptor(RAR)-alpha have been reported. Here, we demonstrate the effects of steroids/thyroids/retinoids and of activators of protein kinase A (forskolin, Forsk) and C (12-O-tetradecanoylphorbol-13-acetate, TPA), on growth and expression of c-erbB and RARs in MCF-7 breast cancer cells, which contain high levels of RAR-alpha and -gamma, and which express significant amounts of c-erbB-2 and -3. All trans-retinoic acid (tRA), the anti-estrogen ICI 182 780 (ICI), Forsk and TPA reduced, whereas triiodothyronine and 17beta-estradiol (E2) stimulated cell growth. Flow cytometry revealed that tRA and E2 reduced c-erbB-2 and -3, whereas tamoxifen, Forsk and TPA up-regulated c-erbB-2. c-erbB-3 was co-regulated with c-erbB-2. Northern analysis demonstrated that RAR-alpha was down-regulated by dexamethasone, ICI, and TPA, whereas vitamin D3 and E2 up-regulated RAR-alpha. RAR-gamma expression was less responsive to such treatment, being reduced only by ICI and Forsk. These data indicate that nuclear receptor and protein kinase signaling communicate with each other and control the expression of RARs and c erbB receptors. Efficient growth control requires the coordinated interplay of both receptor systems. PMID- 10674884 TI - Research potential of a unique xenograft model of human proliferative breast disease. AB - A workshop on the 'Research potential of a unique xenograft model of human proliferative breast disease' was held at the Karmanos Cancer Institute, Wayne State University, Detroit, Michigan, in November of 1998. The accumulated information and current experimental findings on the MCF10AT model of preneoplastic, proliferative breast disease were reviewed. Discussions focused on the relevance of the model to clinical breast cancer and on the most profitable lines of further research to strengthen its utility. PMID- 10674885 TI - Standardization and terminology of nocturia. PMID- 10674886 TI - Lower urinary tract symptoms and nocturia in men and women: prevalence, aetiology and diagnosis. PMID- 10674887 TI - Similarities and dissimilarities between nocturnal enuresis in childhood and nocturia in adults. PMID- 10674888 TI - Nocturia: a disease or normal ageing? PMID- 10674889 TI - Nocturnal polyuria. PMID- 10674890 TI - Measuring the impact of nocturia on quality of life. PMID- 10674891 TI - The impact of sleep deprivation caused by nocturia. PMID- 10674892 TI - Health-economic issues in nocturia. PMID- 10674893 TI - Nocturia: current knowledge and future directions. PMID- 10674894 TI - Clonal diversity of Ig and T-cell receptor gene rearrangements in childhood B precursor acute lymphoblastic leukaemia. AB - The majority of paediatric B precursor acute lymphoblastic leukaemias in children are derived from a single transformed haematopoietic cell with complete or partial VDJ recombination within the immunoglobulin heavy chain gene. A high frequency of patients also show rearrangements within TCRdelta and TCRgamma loci and in up to 40% of children there is an excess of immune system gene rearrangements compared with the number of identified alleles of immune system genes, suggesting the presence of multiple leukaemic subclones -clonal diversity. It has been observed by us and other investigators that in individual patients the pattern of immune system gene rearrangements often changes between presentation and relapse. In order to explore the possibility that clonal diversity plays a biological role during disease progression we optimised methods for subclone detection and analysed the prognostic significance of clonal diversity among 75 children with B precursor-ALL. Our results suggest that clonal diversity plays a role in disease progression as patients with oligoclonal disease showed a significantly shorter disease free survival than patients with monoclonal disease. This trend was of particular importance in the 'standard risk' group of ALL where aggressive disease could not be recognised by other means. In addition, generation of independent subclones from an early, non rearranged tumour progenitor appears to be a common feature among leukaemias with aggressive clinical behaviour. We speculate on the type of genetic factors which may participate both in the generation of subclones and also in wider genomic instability and which are likely to be required for the aggressive clinical phenotype in children with ALL. PMID- 10674895 TI - Chronic lymphoproliferative disorders: an integrated point of view for the differential diagnosis. AB - Morphology is regarded as the principle basis for the identification of lymphoid neoplasms. Sometimes, however, it fails to discriminate among several chronic lymphoproliferative disorders (CLDs). Improved immunophenotyping has resulted in a better characterization of a number of variants of these diseases, some of which may benefit from different therapeutic approaches. In particular, the proposal of scoring systems using a panel of monoclonal antibodies (MoAbs) has represented a critical step in this field. In fact, to date, some MoAbs (CD5, CD23, FMC7, CD22, CD79b, and surface immunoglobulin density) are able to distinguish among several entities, thus allowing for a correct diagnosis in the majority of cases. However, there is still a small percentage of patients where the combined diagnostic approach (morphology and immunophenotyping) should be further refined by other techniques, such as cytogenetic and molecular characterization. Here numerous questions are raised indicating the need to more accurately differentiate the disease entities under discussion and better understand some of their clinical manifestations. PMID- 10674896 TI - Diagnosis and treatment of polycythemia vera and possible future study designs of the PVSG. AB - The present study describes clinicopathological criteria to distinguish the 5 sequential stages proposed by Wasserman et al in the natural history of newly diagnosed PV patients. The European Working Group on MPD (EWG.MPD) extended and modified the PVSG diagnostic criteria of PV by including bone marrow histopathology. From the results of prospective randomized studies in PV it became evident that new clinical trials in previously untreated PV patients should focus on comparing interferon-alpha, a non-leukemogenic approach, versus a potential leukemogenic myelosuppressive treatment modality. Hydroxyurea appears to be the least leukemogenic myelosuppressive agent in long-term prospective clinical PV-studies extending observation periods of more than 10 years. The rational for using IFN-alpha as a first-line treatment option in newly diagnosed PV-patient include its effectiveness to abate constitutional symptoms and to induce a complete remission thereby avoiding phlebotomy, iron deficiency, and macrocytosis associated with hydroxyurea. Moreover IFN-alpha may prevent or delay the development of postpolycythemic myelofibrosis if used early in the course of the disease. Clinicians will be reluctant to postpone the use of hydroxyurea in early stage PV as long as a conservative approach using phlebotomy aiming at a hematocrit below 0.45, plus low-dose aspirin for the control platelet function or anagrelide for the control platelet number is used to keep the patient healthy. Low-dose aspirin will prevent the microvascular thrombotic complications of thrombocythemia associated with PV in remission after phlebotomy, but lacks myelosuppressive activity. Control of megakaryocyte maturation and reduction of platelet production to normal (<400 x 10(9)/l) by relatively low doses of anagrelide will predict a significant reduction of vascular complications in the early stages of PV, may prevent progression to myelofibrosis during follow-up of PV and very probable will postpone the use of hydroxyurea treatment for controlling the platelet count in PV. Large scale randomized clinical trials in PV are proposed, which should aim not only for clinical and hematological response, safety, efficacy, but should also assess toxicity, the need for phlebotomy and whether the development of progressive disease such as splenomegaly, pruritus, myelofibrotic myeloid metaplasia, spent phase, myelodysplasia and acute leukemia can be delayed or prevented by IFN-alpha as compared to a conservative approach of phlebotomy plus low-dose aspirin or anagrelide followed by hydroxyurea when signs of myeloproliferative activity became evident. PMID- 10674897 TI - CD40 and B chronic lymphocytic leukemia cell response to fludarabine: the influence of NF-kappaB/Rel transcription factors on chemotherapy-induced apoptosis. AB - The levels of tumour necrosis factor receptor (TNF-R) superfamily members can be altered in lymphoid leukemias, indicating a possible role of such molecules in the biology of these neoplasias. In B chronic lymphocytic leukemia cells, the CD40/CD40L system has been shown to be effective in inhibiting the apoptotic response to fludarabine. The modulation of apoptosis relied on the CD40-induced activity of NF-kappaB/Rel transcription factors. The anti-apoptotic effect of CD40 was abolished using a phosphorothioate kappaB decoy oligodeoxynucleotide. These findings illustrate an example of the biological activity of TNF-R-like molecules in leukemias. They also show the influence of NF-kappaB/Rel activity on leukemic cell response to apoptogenic agents. PMID- 10674898 TI - Outcome of Philadelphia chromosome-positive adult acute lymphoblastic leukemia. AB - Philadelphia chromosome (Ph)-positive acute lymphoblastic leukemia (ALL) represents the most common cytogenetic abnormality in adult ALL. It is found in 15% to 30% of patients, and its incidence increases with age. As in children, prognosis in Ph-positive adult ALL is poor. No therapeutic approach has had substantial impact on its unfavorable course. We analyzed the characteristics and outcome of newly diagnosed adults with Ph-positive ALL treated at the M. D. Anderson Cancer Center between 1980 and 1997. The diagnosis of patients was based on typical morphological and immunophenotypic criteria of marrow aspirate and biopsy specimens. Cytogenetic and molecular studies were also performed. A total of 67 patients were included in this study. From 1980 until 1991, 38 patients with Ph-positive ALL were treated with vincristine, Adriamycin, and dexamethasone (VAD), or with acute myeloid leukemia (AML)-like induction protocols. Since 1992 a total of 29 patients received induction therapy with an intensified treatment protocol, called "hyper-CVAD". The outcome of patients treated with standard and intensified treatment regimens was compared and results of our institution contrasted with data obtained from other centers. Ph-positive ALL was present in 67 of 498 patients with newly diagnosed ALL (13%). Patients with Ph-positive ALL had a higher median age (44 versus 34, P=0.007), higher median white blood cell (WBC) counts at presentation (25 versus 8, P=0.0002), and higher peripheral median percentage of blast counts (63 versus 40, P=0.023). FAB subtype L2 (70% versus 49%, P=0.001) and CALLA-positive pre-B immunophenotype (75% versus 37%, P<0.001) predominated among Ph-positive ALL. Myeloid marker coexpression was more frequent in Ph-positive ALL when compared with Ph-negative ALL (52% vs. 27% for CD13, P<0.001, and 44% vs. 27% for CD33, P=0.005). Among patients treated with hyper-CVAD, the complete remission (CR) rate was 90% versus 55% (P=0.002) with pre-hyper-CVAD regimens (VAD and AML-like induction protocols), the median CR duration was 43 weeks versus 32 weeks (P>0.5), median disease-free survival (DFS) was 42 weeks versus 29 weeks (P=0.008), and median survival was 66 weeks versus 45 weeks (P>0.5). Patients with hyperdiploid Ph-positive ALL on hyper-CVAD therapy achieved significantly longer CR duration and DFS than hypo- and pseudodiploid cases (59 weeks versus 42 and 31 weeks, P=0.02 and 0.04, respectively). In contrast, patients treated with regimens prior to hyper-CVAD had significantly shorter CR duration (21 weeks versus 33 and 29 weeks, P=0.03) and DFS with hyperdiploid karyotypes when compared to pseudodiploid and hypodiploid cases (16 weeks versus 30 and 13 weeks, P=0.008). In conclusion, our results demonstrate improved response rate and DFS with current intensive regimens (hyper-CVAD) in patients with Ph-positive ALL, but no advantage in overall survival. PMID- 10674899 TI - Aberrant immunophenotypes detected by flow cytometry in acute lymphoblastic leukemia. AB - The present study was designed to analyse the proportion of ALL patients in which the phenotypic detection of minimal residual disease (MRD) is feasible, based on the presence of aberrant phenotypes: lineage infidelity, asynchronous expression, overexpression and ectopic phenotype. For this purpose we have prospectively investigated the phenotype of blast cells from 25 patients at diagnosis using a large panel of monoclonal antibodies by multiparametric flow cytometry. The mean age was 23.3 +/- 17.3 with 10 children and 15 adults. 14 patients were classified as L1, 9 L2 and 2 L3 according to the FAB classification. 17 cases were B-lineage ALL and 8 T-ALL. 23 out of 25 cases (92%) included in this study displayed phenotypic aberrations at diagnosis (15 out of 17 cases of B-lineage ALL and all T-ALL patients). 76% of patients displayed two or more than two aberrancies. The phenotypic aberrations were lineage infidelity, found in 12 patients, asynchronous antigen expression detected in 17 patients, antigen overexpression in 4 patients and ectopic phenotype in 7 patients. In summary our results show that when a large panel of MoAbs is used for the immunophenotypical characterization of ALL, most patients display aberrant phenotypes, the coexistence of more than two aberrant antigen expressions being frequently detected. These results suggest that the use of immunological methods for the detection of MRD in ALL based on the existence of aberrant phenotypes could be of great help for the follow-up of patients in complete remission. PMID- 10674900 TI - Analysis of Wilms tumor gene (WT1) expression in acute leukemia patients with special reference to the differential diagnosis between eosinophilic leukemia and idiopathic hypereosinophilic syndromes. AB - Continuous Wilms' tumor gene (WT1) expression is a typical feature of leukemic blasts in AML, ALL, and blast crisis CML patients. It is easily detectable by a variety of RT-PCR protocols, which differ mainly in their sensitivity. The nuclear WT1 protein can be found in blasts of approximately 50-60% of acute leukemia patients at diagnosis. Conversely, WT1 is only transiently expressed in normal hemopoiesis. Early CD34+ hemopoietic progenitors express WT1, whereas no WT1 mRNA transcripts can be found in mature blood cells and differentiation induced committed CD34- progenitors. As a powerful complementary diagnostic tool, testing for WT1 expression can be helpful to discriminate between eosinophilic leukemia (EoL) patients and patients with idiopathic hypereosinophilic syndromes. Conflicting data about the usefulness of testing for WT1 expression to monitor minimal residual disease (MRD) in treated leukemia patients will be discussed. Finally, research strategies to circumvent shortcomings in detecting leukemia associated WT1 expression will be outlined. PMID- 10674901 TI - Bone marrow features and clinical findings in chronic myeloid leukemia--a comparative, multicenter, immunohistological and morphometric study on 614 patients. AB - A multicenter, immunohistochemical and morphometric study was performed on diagnostic pretreatment bone marrow biopsies in 614 adult patients with Ph1+ chronic myeloid leukemia (CML) to compare histological features with clinical findings. For identification of megakaryopoiesis we used the monoclonal antibody CD61 and additionally the PAS reaction to determine the subfraction of atypical micromegakaryocytes and precursors. Labelling of erythroid precursors was carried out by a monoclonal antibody directed against glycophorin C. In order to selectively stain macrophages and their activated subset we applied CD68 and the GSA-I lectin. Density of argyrophilic fibers (reticulin plus collagen) was measured following Gomori's silver impregnation method. In accordance with laboratory data morphological variables revealed a comparable amount of congruence in the various groups of CML patients derived from different sources. In about 26% of patients early (reticulin) to advanced (collagen) fibrosis was detectable. Significant correlations were calculated between the extent of myelofibrosis with splenomegaly, anemia and increasing numbers of erythroblasts and myeloblasts in the peripheral blood count. These features were assumed to indicate more advanced stages of the disease process with ensuing transition into myeloid metaplasia and consequently were associated with an unfavorable prognosis. Significant relationships were revealed between the number of CD61+ megakaryocytes and more important, also their precursor fraction with the degree of fibrosis. This result extends previous experimental findings regarding the impact of immature elements of this cell lineage for the generation of myelofibrosis. The significant association of erythroid precursors with the number of mature (resident) macrophages including their activated GSA-I subset may shed some light on their functional involvement in iron turnover and hemoglobin synthesis. A modified histological classification of predominant bone marrow features is introduced. This simplified synthesis staging system (Cologne Classification) is not only associated with certain sets of laboratory data, but also with different survival patterns. PMID- 10674902 TI - Diagnostic and clinical implications of lineage fidelity in acute leukemia patients undergoing allogeneic stem cell transplantation. AB - We report on a series of five acute leukemia patients who have undergone allogeneic bone marrow transplantation. Initially, these patients were classified as having biphenotypic leukemia; however, subsequent developments in the perception of what constitutes lineage fidelity has resulted in controversy regarding the diagnosis. Flow cytometry and non-random cytogenetic results have had a major impact on redefining the concept of biphenotypic disease. In this report we review the diagnostic dilemma associated with defining acute leukemia lineage fidelity as diagnostic techniques evolve. While our unique focus on the treatment of biphenotypic leukemia patients represents a small population, we verify the single most promising therapy where otherwise the diagnosis is dismal. PMID- 10674903 TI - Feasibility of high-dose chemotherapy without stem cell support as a first-line treatment for non-Hodgkin's aggressive lymphoma: a pilot study. AB - A regimen which incorporates cyclophosphamide, doxorubicin, vincristine and prednisolone (CHOP) is the standard treatment for patients with non-Hodgkin's lymphoma (NHL), but it has not been effective in patients with aggressive NHL who are at high risk. The aim of the present trial was to investigate the feasibility of high-dose chemotherapy (HDC) without stem cell support as a first-line treatment. The primary endpoint was a complete remission rate. The second endpoint was survival. Fourteen patients with aggressive NHL entered the study and were treated according to the K93 protocol (3 cycles of CHOP, high-dose etoposide and ifosfamide, and high-dose methotrexate) Eleven patients (79%) achieved complete remission (CR) and two (14%) achieved partial remission (PR). Overall survival (OS) after five years was 79%. The actuarial five year disease free survival (DFS) for the eleven cases of CR was 75%. During chemotherapy, grade IV hematologic toxicity was observed in all patients and grade IV non hematologic toxicity in only one patient, who experienced oral ulcers. Peripheral blood stem cell (PBSC) apheresis was performed in eight cases. One harvesting was enough to provide an adequate number of CD34+ cells for the subsequent PBSC transplantation (PBSCT). In conclusion our study confirmed the efficacy of the K93 protocol in obtaining a good response (CR + PR) rate and a very good DFS rate in most cases of aggressive NHL, with acceptable toxicity. This regimen may improve the outcome in cases of aggressive NHL without stem cell support. It seems worthwhile to conduct a randomized controlled study comparing the K93 protocol with the standard CHOP regimen. PMID- 10674904 TI - Feasibility of autologous stem cell transplantation in chronic carriers of hepatitis B and hepatitis C virus. AB - There are several reports describing acute liver decompensation in chronic carriers of HBsAg after withdrawal of chemotherapy or immunosuppressive therapy; recently the same was also reported for chronic HCV-RNA carriers. We retrospectively evaluated hepatic toxicity in eleven patients (6 carriers of HCV RNA and 5 of HBsAg) autotransplanted at our Institution between March '92 and June '98. Male/female ratio was 7/4, median age 41 years (26-56). Nine patients (4 HBsAg) were affected by non-Hodgkin's lymphoma, 1 (HCV-RNA) by chronic myelogenous leukaemia and 1 (HBsAg) by breast cancer. In the immediate post transplant period in only 1 patient (HBsAg carrier and affected by breast cancer) was hepatitis documented (at about 1 month from transplant) with an elevation of transaminase levels (x20-40 n.v.). Neither other complications, nor toxic deaths were observed. During the post-transplant follow-up (median 31 months, range 9 83) no hepatic abnormalities were observed. All patients are alive at 56 months (20-122) from diagnosis. Currently 10/11 patients are in complete remission, while 1 patient, affected by follicular centre lymphoma, is alive with disease 52 months from autologous stem cell transplantation. Our study shows that both conventional therapy and high-dose chemotherapy can be performed safely in chronic hepatitis B and C virus carriers. PMID- 10674905 TI - T-cell receptor gamma and delta gene rearrangements in T-cell acute lymphoblastic leukemia in Indian patients. AB - T-cell acute lymphoblastic leukemia (T-ALL) is a clonal lymphoid malignancy and junctional sequences of rearranged T-cell receptor (TCR) represent the best suitable marker to study clonality in these patients. A sensitive, non radioactive, and rapid approach of PCR coupled with heteroduplex analysis was used to analyse clonality of TCR gamma and delta gene rearrangements in 26 Indian T-ALL patients. Amongst TCR gamma gene family, VgammaI-Jgamma1.3/2.3 sequences were most utilized (53.9%) while from TCRdelta repertoire Vdelta1-Jdelta1 sequences were preferentially rearranged (23.1%) in these patients. 19.2% of Indian T-ALL patients demonstrated both clonal TCR gamma and delta gene rearrangements along with surface expression of TCRgammadelta. Although the majority of T-ALL patients showed surface expression of TCRalphabeta, the small fraction (19.2%) of TCRgammadelta+ T-ALL represent a distinct subgroup which needs further evaluation. PMID- 10674906 TI - Management of severe neutropenia with cyclosporin during initial treatment of Epstein-Barr virus-related hemophagocytic lymphohistiocytosis. AB - Severe neutropenia (absolute neutrophil count <500/gl) is probably due to the combined effects of dysregulated cytokine production and chemotherapeutic agents, and is one of the risk factors in the initial treatment of patients with Epstein Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH). We report here 9 cases of neutropenic HLH, of which 8 were treated with cyclosporin (CSA, 2-6 mg/kg/day; continuous infusion, or 6 mg/kg/day; per os, for periods ranging from 9 days to >8 weeks) in the initial neutropenic phase during induction treatment using corticosteroids and etoposide. Five of the 6 cases, in which CSA treatment was started early (before the second week of induction), survived the critical period with recovery of neutrophil counts within a week. The remaining 3 cases, in which CSA was introduced later or not at all, died of infection. Based on these results, we recommend a prompt short-term CSA infusion during neutropenic episodes in the most common treatment regimen of etoposide and corticosteroids in patients with HLH. Improved neutrophil recovery as a result of CSA treatment makes it possible to continue immunochemotherapy safely and obtain improved patient outcomes. PMID- 10674907 TI - The use of archival bone marrow specimens in detecting B-cell non-Hodgkin's lymphomas using polymerase chain reaction methods. AB - The detection of B-cell non-Hodgkin's lymphoma (B-NHL) involving the bone marrow (BM) can be enhanced by assessing immunoglobulin heavy chain (IgH/JH) gene rearrangement using PCR. While the fresh BM aspirate has been the most commonly used specimen, the utility of archival BM tissues has not been extensively evaluated. We studied the BM from 13 patients with nodal B-NHL (7 low-grade and 6 intermediate grade), which were categorized into three groups based on the histologic finding of lymphoma (H) and the presence of a monoclonal IgH/JH band by PCR using fresh BM aspirates (M): (1) H(+)/M(+), 4 cases; (2) H(+)/M(-), 4 cases; and (3) H(equivocal)/M(-), 5 cases. Archival tissues available for study included paraffin-embedded trephine biopsy (TB)/aspirate clots (AC) and air-dried aspirate smears (AS). All TB (13/13) and a subset of AC (5/13) were B5-fixed, and all these tissues failed to yield analyzable DNA. In contrast, sufficient DNA was consistently obtained in AC that were formalin-fixed (8/13). Of these 8 cases, 2/3 of group 1, 3/3 of group 2, and 0/2 of group 3 had a monoclonal IgH band. Using DNA extracted from microdissected lymphoid aggregates morphologically evident in the AC sections, additional positive cases were identified: 1/3 of group 1 and 2/2 of group 3. In those 5 cases that did not have formalin-fixed TB/AC, sufficient DNA was extracted from AS in all cases; one additional positive case was identified in group 1. Overall, 4/4 (100%) of group 1, 3/4 (75%) of group 2, and 2/5 (40%) of group 3 showed molecular evidence of lymphoma. To conclude, archival BM specimens are a useful source of DNA for molecular detection of B-NHL involvement, and formalin appears to be a better fixative than B5. The use of these samples may improve the overall detection sensitivity. PMID- 10674908 TI - CD5 is A potential selecting ligand for B-cell surface immunoglobulin: a possible role in maintenance and selective expansion of normal and malignant B cells. AB - Although the function of CD5 on B cells is unknown, previous studies suggested that CD5 interaction with V(H) framework regions of surface immunoglobulins (Igs) may contribute to survival and expansion of B cells. Here we used B-chronic lymphocytic leukemia (B-CLL) cells and transformed B-cell lines from normal and B CLL patients to study CD5-Ig interactions. Immobilized Ig binds and permits isolation of CD5 from lysates of CD5-expressing cell lines. Immunoglobulins or Fab fragments of different V(H) families varied in their effectiveness as inhibitors of anti-CD5 staining of CLL cells, appendix and tonsil tissue sections. Human Ig also binds to purified recombinant CD5. We show here for the first time that the unconventional Ig-CD5 interaction maps to the extracellular CD5-D2 domain whereas conventional epitopes recognized by anti-CD5 antibodies are localized in the D1 domain of CD5. We propose that interactions of VH framework regions with CD5 as a ligand may maintain, select or expand normal, autoimmune or transformed B cells and also contribute to skewing of the normal V(H) repertoire. PMID- 10674909 TI - Histamine as an autocrine regulator of leukemic cell proliferation. AB - We examined leukemic lymphocyte precursors from ALL patients as well as immortalized ALL cell lines for cytoplasmic histamine expression. The histamine levels ranged from 10.8 pg/10(6) cells to 82.2 pg/10(6) cells in ALL cell lines (N=4) and from 12.5 pg/10(6) cells to 1235.4 pg/10(6) cells for primary leukemic cells from ALL patients (N=13). The presence of histamine in the cytoplasm of these ALL cells was also confirmed by immunostaining using a polyclonal rabbit anti-histamine antibody. Notably, the histamine receptor blocker diphenhydramine inhibited the clonogenic growth of ALL cells by >90% prompting the hypothesis that histamine may be an autocrine regulator of ALL cell proliferation. Our study suggests that histamine receptor blockers may therefore be useful for the treatment of therapy-refractory ALL. PMID- 10674910 TI - Cell cycle-independent down-regulation of BCL-2 protein expression in differentiating HL-60 cells. AB - To understand the relationship between bcl-2 protein expression and the cell cycle during the processes of differentiation, we examined bcl-2 protein levels expressed during cell cycle phases in differentiating HL-60 human leukemia cells by using a two-color flow cytometric method. In exponentially proliferating HL-60 cells bcl-2 protein was constitutively expressed throughout the cell cycle phases, but a small population of G0/G1 cells expressed decreased levels of protein as compared with other cell cycle phases. HL-60 cells can be induced to differentiate to granulocytic pathway by retinoic acid or dimethylsulfoxide, and to monocytic/macrophagic pathway by 1, 25 dihydroxyvitamin D3 or 12-O tetradecanoylphorbol -13-acetate. During treatment with any of these inducing agents, bcl-2 protein expression was time-dependently down-regulated after 24 h. A two-color flow cytometric analysis revealed that this down-regulation occurred throughout cell cycle phases, indicating that bcl-2 protein expression was down regulated in cell cycle-independent manner during induction of differentiation in HL-60 cells. To our knowledge, this is the first demonstration suggesting that the regulation of bcl-2 protein expression is not related to the cell cycle during induction of differentiation in HL-60 cells. PMID- 10674911 TI - Light microscopic detection of BCR-ABL transcripts after in-cell RT-PCR: fusion gene expression might correlate with clinical evolution of chronic myeloid leukemia. AB - A procedure for in-cell amplification of the hybrid BCR-ABL mRNA by reverse transcription and polymerase chain reaction (RT-PCR) without extraction of the nucleic acids was performed directly in fixed and permeabilized cells of leukemia patients (22 patients with Ph'-positive chronic myeloid leukaemia-CML and 1 with Ph'-positive acute leukaemia-AL, as well as 7 Ph'-negative cases) and Ph' positive human leukaemia cell lines (K562, LAMA-84, BV173). The labelling of the amplified sequences was done employing biotinylated primers and a second PCR in a semi-nested fashion with a low number of cycles. An enzymatic system based on biotin-streptavidin-chromogen reaction was used for the detection of labeled PCR product, thus producing a coloured product, visible to the eye under a standard light microscope. All samples from patients with cytogenetic and molecular evidence of BCR-ABL rearrangement showed specific cytoplasmic staining at the site of the amplified hybrid transcripts. It allowed definite distinction between positive and negative cells. K562, LAMA-84, BV173 cells were characterized with strong diffuse staining while an interesting finding of the present study was the presence of variable quantities of colour product in patients' samples which might be due to different mRNA expression. Early and intermediate stages of myeloid maturation showed more intense reactivity. Cases with an aggressive course of accelerated or blast phase CML and AL were found to have a considerable subset of cells with strongly expressed signal while cases in chronic phase were characterised with uniform weak to moderate reaction. Our observations support the hypothesis that the amount of BCR-ABL transcript expression within neoplastic cells may play a role in dictating the eventual behaviour of the leukaemic clone. Future studies at a single cell level of larger series of consecutive cases with a follow up might be able to identify those patients who are prone to transformation and provide certain indications for further therapeutic decisions. PMID- 10674912 TI - HIV infection of megakaryocytic cell lines. AB - Thrombocytopenia is a common hematologic disorder in HIV infection and occurs in both asymptomatic and AIDS patients. An autoimmune mechanism has been postulated for the platelet destruction associated with some forms of thrombocytopenia. However, recent studies revealed that megakaryocytes are susceptible to HIV infection and suggested the possibility that HIV can directly impair the platelet production from megakaryocytes. This study was designed to characterize the HIV receptor expression in megakaryocytic cells and the responsiveness to HIV infection. Four different megakaryocytic cell lines at different stages of differentiation were established from the peripheral blood of different individuals with hematologic malignancies. CMK and CMY cells (differentiated cell lines) expressed CD4, but CMS and CTS cells (poorly differentiated cell lines) did not. The HIV coreceptor CXCR4 was also expressed in CMY and CMK cells. HIV-1 (HTLV-IIIB) replicated in CMY cells persistently but not in other three cell lines. CMY cells as well as CMK cells were also susceptible to the lytic infection of HIV-2 (LAV2). Pretreatment of the CMY cells with anti-CD4 antibody inhibited the infection by both HIV-1 and HIV-2. Our results indicate that mature megakaryocytic cells express CD4 along with HIV coreceptors and are susceptible to HIV infection. PMID- 10674913 TI - Temporary remission of an alveolar rhabdomyosarcoma diagnosed and treated as acute leukemia. AB - A 29-year-old man with alveolar rhabdomyosarcoma was considered to be suffering from acute leukemia. A bone marrow aspirate had revealed extensive infiltration by atypical blast-like cells which were interpreted as acute lymphoblastic leukemia. Although there was no confirmation of this diagnosis by immunophenotyping chemotherapy with a protocol suited for the treatment of acute lymphoblastic leukemia was started prior to histological analysis and resulted in a complete temporary remission after the first cycle. Histological analysis of a bone marrow biopsy revealed an alveolar rhabdomyosarcoma, as further confirmed by molecular genetic analysis. Two months after the end of chemotherapy, there was an extensive recurrence and the patient died one year after initial diagnosis with chemotherapy refractory disease. In conclusion, rhabdomyosarcoma should always be included in the differential diagnosis of systemic diseases with extensive bone marrow infiltration by tumor cells which could otherwise be misinterpreted as a haematological malignancy. PMID- 10674914 TI - Epstein-Barr virus-associated high-grade anaplastic plasmacytoma in a renal transplant patient. AB - Allograft transplant patients have an increased risk of developing polyclonal or monoclonal lymphoproliferative disorders, but high-grade anaplastic plasmacytomas are extremely rare in these patients. We present a renal transplant patient who developed multiple extramedullary high-grade anaplastic plasmacytomas in the oral cavity, the left maxillary antrum, the scalp, the thigh and the upper abdominal wall with no evidence of diffuse bone marrow infiltration. Epstein-Barr virus (EBV) mRNA transcripts were detected within the myeloma cells by in situ hybridization using EBER1-2 probes. Following discontinuation of immunosuppression applied, the patient was treated with a cyclophosphamide prednisone regimen followed by local irradiation, and a complete remission was achieved within four weeks. We concluded that EBV-associated high-grade anaplastic plasmacytomas constitute one more type of post-transplant lymphoproliferative disorder, and that despite their characterization as highly malignant neoplasms, their clinical behavior is not always aggressive. PMID- 10674915 TI - Postmastectomy malignant lymphoma. AB - Postmastectomy lymphedema (PML) is a morbid condition occurring in patients with breast carcinoma treated with radical/modified radical mastectomy. Postmastectomy angiosarcoma (PMA) is the most common neoplasia seen in these patients. Primary malignant lymphoma arising in PML is a rare neoplasia and 3 cases have been reported until now. In this report a patient with diffuse large cell lymphoma (DLCL) arising in PML is reported and the other three cases are reviewed. PMID- 10674916 TI - Non-Hodgkin's lymphoma following untreated essential thrombocythemia. AB - The coexistence of essential thrombocythemia (ET) and non-Hodgkin's lymphoma (NHL) is an extremely rare event, with only two such cases having been reported in the medical literature. We describe here a 25-year old woman who developed high-grade B-cell NHL of the stomach three years after the diagnosis of ET, for which she had received no treatment, due to her young age and the lack of thrombotic risk factors other than thrombocytosis. The lymphoma showed a favorable response to CHOP chemotherapy, whereas the thrombocytosis remained unchanged throughout the patient's clinical course. The possible etiologic and pathogenetic mechanisms leading to the association of these two disorders are discussed. Given the relative frequency of ET and the fact that the present case represents only the third reported instance of NHL developing in such patients, the coincidental ocurrence of both diseases is a possibility that cannot be excluded. PMID- 10674917 TI - Inducible types of cyclooxygenase and nitric oxide synthase in adaptive cytoprotection in rat stomachs. AB - Roles of cyclooxygenases (COX-1 and COX-2) and nitric oxide (NO) synthases (nNOS and iNOS) in adaptive cytoprotection induced by 20 mM taurocholate dissolved in 50 mM HCl (TC) were investigated in rat stomachs. Intragastric administration of 0.6 N HCl caused haemorrhagic damage in the stomach. These lesions were prevented by pretreatment of the animals with TC p.o. 0.5 h before 0.6 N HCl, and a significant protection persisted for more than 5 h. The protection afforded by TC given 0.5 h before HCl was almost totally reversed by indomethacin and slightly mitigated by N(G)-nitro-L-arginine methyl ester (L-NAME) but not affected by NS 398 or aminoguanidine. By contrast, the mucosal protective action of TC given 5 h before HCl was significantly reversed by NS-398, L-NAME and aminoguanidine as well as indomethacin. Mucosal prostaglandin E2 (PGE2) contents were significantly increased for over 5 h after TC, while luminal NOx output tended to elevate at 0.5 h and be significantly increased at 5 h after TC. The increased PGE2 generation observed 0.5 h after TC was attenuated only by indomethacin, while that observed 5 h after TC was inhibited by NS-398 as well as indomethacin. On the other hand, the NOx output determined at 5 h after TC was significantly reduced by both L-NAME and aminoguanidine. The expression of mRNA for both COX-2 and iNOS was apparently detected in the stomach from 3 h after TC treatment. These results suggest that TC induced adaptive cytoprotection in the rat stomach against 0.6 N HCl, the effect lasting for over 5 h, and the underlying mechanism differs depending on the period after the irritation. The early phase is mediated mainly by COX-1/PGs, while the later phase is mediated by iNOS/NO, in addition to prostaglandins (PGs) produced by both COX-1 and COX-2. PMID- 10674918 TI - The role of the host versus the environment in duodenal ulcer disease. AB - Gastric functions can be understood only in the context of a network including the brain gut axis, neuro-endocrine and paracrine mechanisms and growth factors. These host factors including parietal cell sensitivity (PCS) may well interact with an important environmental factor, Helicobacter pylori (Hp), and help to explain its actions. The aim of this study was to investigate PCS related to Hp status and duodenal ulcer (DU). PCS was assessed by constructing dose-response curves after pentagastrin and calculating the D50. Five groups of patients were studied: I) active DU, Hp pos. (8); II) history of DU, Hp pos. (8); III) asymptomatic Hp pos. (8); IV) asymptomatic Hp neg. (10); V) DU on maintenance H2 blocker therapy, Hp pos. (20). PCS was repeated after Hp eradication. PCS was lowest in group IV, and in Hp pos. groups, was significantly higher, with insignificant differences among them, irrespective of DU. PCS declined significantly after Hp eradication. Group V showed an insignificant decline in PCS during treatment, not preventing recurrence. A higher PCS in Hp infection irrespective of DU, declining after eradication, suggests that this may be a reversible epiphenomena related to Hp infection. This may offer an explanation as to why DU develops only in some subjects with Hp, suggesting the importance of the host in the pathogenesis of DU. PMID- 10674919 TI - Cytoprotection and intracellular calcium. AB - It seems that prostacyclin has an increasing effect on gastric mucosal (antral and fundic) calmodulin level in rats. Using either the calcium channel blocker verapamil or anti-calmodulin drugs (diazepam, trifluoperazine,) the cytoprotective effect of prostacyclin can be inhibited. Therefore, it is probable that calcium ions and calcium-activated calmodulin play a role in the effect of prostacyclin. PMID- 10674920 TI - Roles of endogenous prostaglandins and nitric oxide in gastroduodenal ulcerogenic responses induced in rats by hypothermic stress. AB - We examined the roles of endogenous prostaglandins (PGs) and nitric oxide (NO) in the gastroduodenal ulcerogenic responses to hypothermic stress (28 approximately 30 degrees C) in anesthetized rats. Lowering body temperature provoked damage in the gastroduodenal mucosa, with an increase of gastric acid secretion and motility. These responses were completely abolished by bilateral vagotomy or atropine, while 16,16-dimethyl PGE2 decreased the mucosal ulcerogenic response with no effect on acid secretion. The non-selective COX inhibitors, indomethacin or aspirin, worsened these lesions with enhancement of gastric motility and no effect on acid secretion, while the selective COX-2 inhibitor NS-398 did not affect any of these responses. On the other hand, the non-selective NOS inhibitor L-NAME but not aminoguanidine (a relatively selective inhibitor of iNOS), significantly potentiated the acid secretory and mucosal ulcerogenic responses in the stomach but reduced the duodenal damage in response to hypothermia, the effects being antagonized by co-administration of L-arginine. Hypothermia itself decreased duodenal HCO3- secretion under both basal and mucosal acidification stimulated conditions. Both indomethacin and aspirin further decreased the HCO3- response to the mucosal acidification, while L-NAME significantly increased the HCO3- secretion even under hypothermic conditions, similar to 16,16-dimethyl PGE2. These results suggest that 1) hypothermic stress caused an increase of acid secretion and motility as well as a decrease of duodenal HCO3-secretion, resulting in damage in both the stomach and duodenum, 2) the COX-1 but not COX-2 inhibition worsened these lesions by enhancing gastric motility and further decreasing duodenal HCO3- response, 3) the cNOS but not iNOS inhibition worsened gastric lesions by increasing acid secretion but decreased duodenal damage by increasing HCO3- secretion. Thus, it is assumed that the gastroduodenal ulcerogenic and functional responses to hypothermic stress are modified by cNOS/NO as well as COX-1/PGs. PMID- 10674921 TI - Small doses of capsaicin given intragastrically inhibit gastric basal acid secretion in healthy human subjects. AB - Although the direct inhibitory effect of small dose of capsaicin on gastric secretory responses was proved in animal observations, the role of capsaicin sensitive afferent nerves (CSAN) and the effect of capsaicin applied in small and high doses on gastric secretion in human has not been clarified yet. In this study we investigated the influence of different small doses (100-800 microg) of capsaicin given intragastrically through an orogastric tube on gastric basal secretory responses in 10 healthy human subjects. Gastric basal secretory responses (volume, H+-concentration, H+-output) were measured from the suctions of gastric juice for a 1-h period. It has been found that: a) capsaicin dose dependently inhibited the volume and H+-output of gastric juice; b) ID50 was found to be about 400 microg for capsaicin on gastric acid secretion; c) the time interval for capsaicin-induced gastric inhibition existed for about 1 h indifferently from the higher dose (800 microg) of capsaicin given after. It has been concluded that the capsaicin (given in small doses) inhibits the gastric basal acid output via stimulation of the inhibition of capsaicin sensitive afferent nerves. PMID- 10674922 TI - A novel method to produce extensive gastric antral ulcer in rats: pharmacological factors involved in the etiology of antral ulceration. AB - Gastric antral area is the most susceptible region to gastric ulceration in man. However, only limited information is available on animal models. In the present paper, we have developed an improved method for inducing gastric antral ulcers by the administration of 1.0 M HCl after refeeding for 1 h in rats. On day 4, the severe ulcer was found covering extensively the whole area of the antrum, and penetrated through the muscularis mucosae. The incidence of ulceration was 100% and the mean ulcer index was 37.1 +/- 16.6 mm2. In contrast, none of the erosive lesions were observed in the corpus area. Before 24 h, only slight hyperemia was observed in the antral region, suggesting that some submucosal mechanisms are involved in the ulceration processes other than the direct erosive action of HCl on the mucosal surface. Additional treatment with diethyldithiocarbamate (125 mg x kg(-1), s.c.), superoxide dismutase inhibitor, significantly aggravated this antral ulcer, and the ulcer index was 66.0 +/- 13.6 mm2. Allopurinol (50 mg x kg( 1), p.o.) significantly prevented ulcer formation induced by HCl plus DDC. GSH (150 mg x kg(-1), i.p.) also markedly prevented the ulceration. However, DMSO (0.5%, 5 mL x kg(-1), p.o.) was found not to affect ulcer formation. Famotidine (20 mg x kg(-1), p.o.) almost completely inhibited ulcer formation. From the above results, it was concluded that gastric antral ulcer can be induced by the simple treatment of 1.0 M HCl in refed rats, and the antrum has a different defensive mechanism from that in the corpus area. In addition. oxygen derived radicals, especially superoxide anion and endogenous acid secretion were found to be involved in the etiology of the aggravation of the gastric antral ulcer induced by DDC. PMID- 10674923 TI - Capsaicin-sensitive afferent sensory nerves in modulating gastric mucosal defense against noxious agents. AB - In the rat stomach, evidence has been provided that capsaicin-sensitive sensory nerves (CSSN) are involved in a local defense mechanism against gastric ulcer. In the present study capsaicin or resiniferatoxin (RTX), a more potent capsaicin analogue, was used to elucidate the role of these sensory nerves in gastric mucosal protection, mucosal permeability, gastric acid secretion and gastrointestinal blood flow in the rat. In the rat stomach and jejunum, intravenous RTX or topical capsaicin or RTX effected a pronounced and long lasting enhancement of the microcirculation at these sites, measured by laser Doppler flowmetry technique. Introduction of capsaicin into the rat stomach in very low concentrations of ng-microg x mL(-1) range protected the gastric mucosa against damage produced by topical acidified aspirin, indomethacin, ethanol or 0.6 N HCl. Resiniferatoxin exhibited acute gastroprotective effect similar to that of capsaicin and exerted marked protective action on the exogenous HCl, or the secretagogue-induced enhancement of the indomethacin injury. The ulcer preventive effect of both agents was not prevented by atropine or cimetidine treatment. Capsaicin given into the stomach in higher desensitizing concentrations of 6.5 mM markedly enhanced the susceptibility of the gastric mucosa and invariably aggravated gastric mucosal damage evoked by later noxious challenge. Such high desensitizing concentrations of capsaicin, however, did not reduce the cytoprotective effect of prostacyclin (PGI2) or beta-carotene. Capsaicin or RTX had an additive protective effect to that of atropine or cimetidine. In rats pretreated with cysteamine to deplete tissue somatostatin, capsaicin protected against the indomethacin-induced mucosal injury. Gastric acid secretion of the pylorus-ligated rats was inhibited with capsaicin or RTX given in low non-desensitizing concentrations, with the inhibition being most marked in the first hour following pylorus-ligation. Low intragastric concentrations of RTX reduced gastric hydrogen ion back-diffusion evoked by topical acidified salicylates. It is concluded that the gastropotective effect of capsaicin-type agents involves primarily an enhancement of the microcirculation effected through local release of mediator peptides from the sensory nerve terminals. A reduction in gastric acidity may contribute to some degree in the gastric protective action of capsaicin-type agents. The vasodilator and gastroprotective effects of capsaicin-type agents do not depend on vagal efferents or sympathetic neurons, involve prostanoids, histaminergic or cholinergic pathways. PMID- 10674924 TI - Capsaicin increases gastric emptying rate in healthy human subjects measured by 13C-labeled octanoic acid breath test. AB - The role of capsaicin-sensitive primary afferent sensory nerves in the regulation of gastrointestinal motility in human is not clarified yet. In this study, we investigated the effect of 400 microg capsaicin given intragastrically on gastric emptying measured by 13C-octanoic acid breath test in ten healthy human subjects. Four parameters of gastric emptying curves were taken into consideration: 1) maximum value of the curve, 2) time belonging to this maximum, 3) slope of the rising part of the curve and 4) time belonging to the 50% of the area under the curve. Administration of 400 microg capsaicin significantly increased the slope of gastric emptying curve (from 0.1 +/- 0.01 to 0.139 +/- 0.014 U x min(-1), P < 0.05) and significantly decreased the time belonging to the maximum value of emptying curve (from 150 +/- 18 to 75 +/- 12 min, P < 0.05) and the time belonging to the 50% of the area under the curve (from 112 +/- 15 to 99 +/- 14 min, P < 0.05). According to our results 400 microg capsaicin enhances gastric emptying rate in healthy human subjects. PMID- 10674925 TI - Capsaicin inhibits the pentagastrin-stimulated gastric acid secretion in anaesthetized rats with acute gastric fistula. AB - The effect of capsaicin on basal and pentagastrin-stimulated gastric acid secretion was investigated in the urethane anaesthetized acute gastric fistula rat. Gastric acid secretion was measured by flushing of the gastric lumen with saline every 15 min or by continuous gastric perfusion. Capsaicin given into the rat stomach at 120 ng x mL(-1) prior to pentagastrin (25 microg x kg(-1), iv) reduced gastric acid secretory response to pentagastrin by 24%. Intravenous (iv) capsaicin (0.5 microg x kg(-1)) did not reduce the pentagastrin-stimulated gastric acid secretion. After topical capsaicin desensitization (3 mg x mL(-1)), basal gastric acid secretion and that in response to pentagastrin (25 microg x kg(-1), intraperitonaeally) was unaltered compared with the control group. Data indicate that topical capsaicin inhibits gastric acid secretion stimulated with pentagastrin in anaesthetized rats. PMID- 10674926 TI - Influence of column design on process-scale ion-exchange chromatography. AB - The performance of two new designs of pump-packed axial flow process chromatography columns have been evaluated for the preparative anion-exchange chromatography of hen egg-white proteins using Whatman Express-Ion Exchanger Q. A 16 1 Side-Pack column and a 24 1 IsoPak column containing Express-Ion Q were used in this study. In each case ca. 20 1 feedstock containing 5-7 g protein/l, was applied per litre packed bed at flow-rates of ca. 150 and 300 cm/h. In each case the ovalbumin binding capacity was ca. 70 g/l packed bed with ca. 100% (w/w) recovery of applied protein. A clean-in-place procedure involving storage in 0.5 M NaOH was effective in maintaining chromatographic performance in all cases. These data were consistent with our previous work using the more traditionally configured slurry-packed axial flow columns. Each of these column designs were easy to use facilitating rapid packing with this adsorbent and in the case of IsoPak rapid pump unpacking. The introduction of these column designs significantly improves the task of column packing, hitherto a labour intensive, physically demanding and potentially unreproducible process. PMID- 10674927 TI - Modeling and simulation of the dynamic behavior of monoliths. Effects of pore structure from pore network model analysis and comparison with columns packed with porous spherical particles. AB - A mathematical model is presented that could be used to describe the dynamic behavior, scale-up, and design of monoliths involving the adsorption of a solute of interest. The value of the pore diffusivity of the solute in the pores of the skeletons of the monolith is determined in an a priori manner by employing the pore network modeling theory of Meyers and Liapis [J. Chromatogr. A, 827 (1998) 197 and 852 (1999) 3]. The results clearly show that the pore diffusion coefficient, Dmp, of the solute depends on both the pore size distribution and the pore connectivity, nT, of the pores in the skeletons. It is shown that, for a given type of monolith, the film mass transfer coefficient, Kf, of the solute in the monolith could be determined from experiments based on Eq. (3) which was derived by Liapis [Math. Modelling Sci. Comput., 1 (1993) 397] from the fundamental physics. The mathematical model presented in this work is numerically solved in order to study the dynamic behavior of the adsorption of bovine serum albumin (BSA) in a monolith having skeletons of radius r(o) = 0.75x10(-6) m and through-pores having diameters of 1.5x10(-6)-1.8x10(-6) m [H. Minakuchi et al., J. Chromatogr. A, 762 (1997) 135]. The breakthrough curves of the BSA obtained from the monolith were steeper than those from columns packed with porous spherical particles whose radii ranged from 2.50x10(-6) m to 15.00x10(-6) m. Furthermore, and most importantly, the dynamic adsorptive capacity of the monolith was always greater than that of the packed beds for all values of the superficial fluid velocity, Vtp. The results of this work indicate that since in monoliths the size of through-pores could be controlled independently from the size of the skeletons, then if one could construct monolith structures having (a) relatively large through-pores with high through-pore connectivity that can provide high flow-rates at low pressure drops and (b) small-sized skeletons with mesopores having an appropriate pore size distribution (mesopores having diameters that are relatively large when compared with the diameter of the diffusing solute) and high pore connectivity, nT, the following positive results, which are necessary for obtaining efficient separations, could be realized: (i) the value of the pore diffusion coefficient, Dmp, of the solute would be large, (ii) the diffusion path length in the skeletons would be short, (iii) the diffusion velocity, vD, would be high, and (iv) the diffusional response time, t(drt), would be small. Monoliths with such pore structures could provide more efficient separations with respect to (a) dynamic adsorptive capacity and (b) required pressure drop for a given flow-rate, than columns packed with porous particles. PMID- 10674928 TI - Polycations as displacer in high-performance bioseparation. AB - Displacement chromatography is an interesting but up to now rarely used type of preparative biochromatography. The lack of well-engineered and accessible displacer contributes to this phenomenon. In this paper a novel type of displacer is introduced for cation-exchange displacement chromatography, which will soon become commercially available. The molecule is a well-defined PolyDADMAC [poly(diallyldimethylammonium chloride)] with a molar mass of less than 35000 g/mol, an exclusively linear structure and a molar mass polydispersity of less than 1.5. A method for synthesizing such a polymer at high yields is described. The PolyDADMAC is shown to be an efficient displacer of basic proteins from strong cation-exchange columns. PMID- 10674929 TI - Development of an high-performance liquid chromatographic simulated moving bed separation from an industrial perspective. AB - A binary test mixture consisting of cyclopentanone and cycloheptanone is used for the performance evaluation of a pilot-scale simulated moving bed unit. The involved adsorption equilibria and the kinetic behavior are discussed in detail. The results of the test runs are evaluated using the recently introduced "triangle theory" which allows to account for the overload conditions prevailing under preparative chromatographic conditions and to select optimal operating conditions. Under optimized conditions the separation of 735 g test mixture/kg stationary phase per day with purities >99.9% for extract and raffinate stream has been achieved. PMID- 10674930 TI - Quantification of single solute and competitive adsorption isotherms using a closed-loop perturbation method. AB - A modification of the classical method of perturbation chromatography for measuring isotherms of the adsorption of dissolved components is suggested. The general principle of the method consists in analyzing responses of the chromatographic system to small perturbations at different equilibrium concentrations. Essential advantages of the method are: (a) only retention times or volumes have to be measured and no detector calibration is required and (b) experiments with mixtures can be performed and analyzed efficiently. The modification suggested in this paper is the application of a closed-loop arrangement allowing the efficient exploitation of the sample. Experimental data for four different chromatographic systems are presented to illustrate the method. With the determined adsorption isotherms elution profiles could be predicted satisfactorily. PMID- 10674931 TI - Visualization of viscous fingering in high-performance liquid chromatographic columns. Influence of the header design. AB - Using an on-column visualization technique, band profiles of solutes migrating along an HPLC column were studied. The study showed that, under conditions where viscous fingering is prevalent, the design of the inlet header has little influence on the outcome of the viscous fingers. Two types of headers were studied. The first contained a small diameter inlet frit, which localized the majority of the sample in or near the central region of the column. The second header contained a wide frit and produced a more uniform radial distribution of the sample. In both cases, the extent of viscous fingering was essentially the same. PMID- 10674932 TI - Visualization of sample introduction in liquid chromatographic columns. Contribution of a flow distributor on the sample band shape. AB - The contributions to the radial distribution of the sample concentration across the column inlet and to the axial band dispersion resulting from a column header containing a distributor were evaluated using a band-visualization process entailing matching the refractive indices of the stationary and mobile phases in a glass column. This study illustrates graphically how a distributor fitted to the column can increase the axial dispersion of the sample band compared to an inlet containing only a frit. The distributor did not provide a uniform sample distribution across the column. In fact, for 17-mm inner diameter columns and high-porosity frits, the distribution was no better than with the frit having no distributor. However, when low-porosity frits were employed, improved peak shapes were obtained with a distributor. In addition, we observed that the inlet header configuration influenced dramatically the flow stream established along the column. The radial distribution of the efficiency of the columns was nearly homogeneous for those having only a frit but not for those having also a distributor. For the latter, the efficiency decreased from the column axis to its wall. PMID- 10674933 TI - Determination of the thermodynamic contribution to peak asymmetry of basic solutes in reversed-phase liquid chromatography. AB - To this day packing materials manufacturers are still trying to develop reversed phase stationary phases that have silica more completely reacted with bonding ligands to afford more homogeneous particle surfaces. Incomplete bonding causes inhomogeneous effects that are readily observed when separating basic solutes because of the acidic silanols that are unreacted. However, it is still not understood exactly what types of silanol sites are unreacted or if metal impurities are contributing to the resulting peak asymmetry observed. A method is presented which utilizes (1) the frontal analysis method of chromatography to obtain adsorption/partition isotherms, (2) a heterogeneous Langmuir distribution model for the resulting isotherm, (3) an expectation-maximization numerical procedure to solve the mathematical problem to yield the most probable distribution of adsorption parameters, and (4) the equilibrium-dispersive model of chromatography incorporating the fitted isotherm model to check the validity of the sorption model with experimental observations. Correlation of packing materials characterization parameters with results obtained by this procedure will indicate what type of silanols or other sites are responsible for observed tailing behavior. Developers and manufacturers will then be able to more efficiently target their synthetic designs for "base-deactivated" reversed-phase silicas. PMID- 10674934 TI - Study of hydrophobic interaction adsorption of bovine serum albumin under overloaded conditions using flow microcalorimetry. AB - The adsorption behavior of bovine serum albumin (BSA) on a Sepharose based hydrophobic interaction support has been studied. Flow microcalorimetry has been used to determine the heat of adsorption under overloaded chromatographic conditions. These data have been complemented with capacity factor and isotherm measurements to provide insight on the mechanisms of adsorption. The heat of adsorption data have confirmed that the hydrophobic interaction adsorption of BSA under linear isotherm conditions is driven by entropy changes. Under overloaded (non-linear) conditions, however, it has been shown that the changes in enthalpy can drive adsorption; this behavior is not evident from analyses of capacity factor data. It is postulated that for BSA adsorption on the Sepharose derivative of interest, attractive force interactions between adsorbed protein molecules drive the adsorption process under overloaded conditions in a high (NH4)2SO4 environment. It is further postulated that these interactions are due to a change in confirmation of the adsorbed protein under these conditions. PMID- 10674935 TI - Studies on the chromatographic fractionation of Trichoderma reesei cellulases by hydrophobic interaction. AB - This work reports new studies on cellulases fractionation by hydrophobic interaction chromatography. The purification procedure for the Trichoderma reesei cellulase complex consists of gel permeation chromatography on Sephadex G-25M followed by an ultrafiltration step. The concentrated enzyme solution was then fractionated on Sepharose CL-6B modified by covalent immobilization of 1,4 butanediol diglycidyl ether. The influence of the mobile phase composition on the chromatographic behaviour of the T. reesei cellulase complex was investigated. By using 13% (w/v) ammonium sulphate in eluent buffer, a selective separation of beta-glucosidase with a two-fold increase in specific activity and a recovery of 60% cellobiase activity were obtained. Other commercial hydrophobic supports (octyl- and phenyl-Sepharose) were also tested and compared under the same conditions. PMID- 10674936 TI - Expanded-bed chromatography in primary protein purification. AB - Chromatography in stable expanded beds enables proteins to be recovered directly from cultivations of microorganisms or cells and preparations of disrupted cells, without the need for prior removal of suspended solids. The general performance of an expanded bed is comparable to a packed bed owing to reduced mixing of the adsorbent particles in the column. However, optimal operating conditions are more restricted than in a packed bed due to the dependence of bed expansion on the size and density of the adsorbent particles as well as the viscosity and density of the feedstock. The feedstock composition may become the most limiting restriction owing to interactions of adsorbent particles with cell surfaces, DNA and other substances, leading to their aggregation and consequently to bed instabilities and channeling. Despite these difficulties, expanded-bed chromatography has found widespread applications in the large scale purification of proteins from mammalian cell and microbial feedstocks in industrial bioprocessing. The basics and implementation of expanded-bed chromatography, its advantages as well as problems encountered in the use of this technique for the direct extraction of proteins from unclarified feedstocks are addressed. PMID- 10674937 TI - Electrophoretic transfer of proteins across polyacrylamide membranes. AB - The electrophoretic transfer of purified proteins has been examined in a Gradiflow "Babyflow BF100" unit. A number of factors affect protein separation within this preparative electrophoresis system. We established that the rate of protein transfer was proportional to the applied voltage. The transfer is slowest at the isoelectric point (pI) and increased the further away the pH was from the pI of the protein. Protein transfer was found to be independent of the ionic strength of the buffer, for buffers that excluded the addition of strong acids or strong bases or sodium chloride. Transfer decreased as the pore size of the membrane decreased. Finally, transfer was inhibited at high salt concentrations in the protein solution, but remained unaffected when urea and non-ionic detergents were added to the solution. To increase the speed of protein separations, buffers with low conductivity should be used. A pH for the optimal separation should be selected on the basis of the relative pI and size of the target proteins and that of the major contaminants. PMID- 10674938 TI - Protein diffusion in charged polyacrylamide gels. Visualization and analysis. AB - Protein diffusion in anionic, cross-linked polyacrylamide-based gels supported in fused-silica capillaries was characterized by a direct visualization method. Microphotography was used to obtain transient protein concentration profiles in these gels using cytochrome c as a probe molecule. Gels based on acrylamido methylpropane sulfonic acid with 2.5-10% N,N'-methylene-bisacrylamide as a cross linker and with a total polymer concentration of 0.21 g/cm3 yielded diffuse protein concentration profiles which were quantitatively consistent with a Fickian diffusion model. An analytical method was developed to calculate the diffusivity as a function of protein concentration in the gel from the experimental profiles. The diffusivity was found to assume values in the range 2.5-5.5x10(-8) cm2/s and varied somewhat with the protein concentration in the gel. The effects of some of the polymer properties, such as cross-link density, polymer concentration and charge, were also investigated for a limited range of conditions to derive qualitative trends. Results showed that the transport rates increased with a decrease in the cross-link density, were extremely reduced when the polymer concentration was doubled, and were slightly increased when the charge density was decreased by half by polymerizing a 1:1 mixture of acrylamide and acrylamido-methylpropane sulfonic acid monomers. PMID- 10674939 TI - Rapid reversed-phase separation of proteins and peptides using optimized 'moulded' monolithic poly(styrene-co-divinylbenzene) columns. AB - Monolithic macroporous poly(styrene-co-divinylbenzene) stationary phases have been prepared by free radical polymerization within the confines of 4.6-mm I.D. chromatographic columns. The optimized porous properties allow the mobile phase to flow through these columns at flow-rates of up to 10 ml/min. As opposed to the simultaneously tested columns packed with either silica or synthetic polymer beads, the monoliths exhibit only modest back pressure. The monolithic columns were able to separate mixtures of peptides and proteins in a very short time. Under the optimized conditions, the separation of five proteins can be easily achieved in less than 20 s. PMID- 10674940 TI - Separation of stereoisomers in a simulated moving bed-supercritical fluid chromatography plant. AB - The combination of two techniques, simulated moving bed (SMB) and supercritical fluid chromatography (SFC), leads to an apparatus with unique features. Besides the known advantages of the SMB process, like reduced solvent consumption and its continuity, the use of supercritical carbon dioxide as the mobile phase offers an easy product recovery by depressurizing the supercritical fluid. Details of a SMB SFC plant are presented for the first time. Due to the large number of process parameters a simulation of the SMB process is necessary to achieve optimal operating conditions. The most important thermodynamic information for a SMB process is the adsorption isotherms. Therefore, isotherms for two phytol isomers are measured and correlated. A fast dynamic model for the simulation of SMB is used to calculate the region of complete separation taking different column configurations and the compressibility of the mobile phase into account. PMID- 10674941 TI - Enantiomers separation by simulated moving bed chromatography. Non-instantaneous equilibrium at the solid-fluid interface. AB - The simulated moving bed (SMB) technology, first conceived for large bulk-scale separations in the petrochemical industry, has found increasingly new applications in the pharmaceutical industry. Among these, the separation of fine chemicals has been the subject of considerable study and research. This work presents the modeling, simulation and design of the operation of a SMB plant in order to separate a binary chiral mixture. The usual assumption of instantaneous equilibrium at the solid-fluid interface is questioned and a first-order kinetics of adsorption is taken into account. The cases of linear, Langmuir and modified Langmuir equilibria are studied. The equivalent true moving bed (TMB) model was used assuming axial dispersion for the fluid flow and plug flow for the solid phase flow. Intraparticle diffusion was described by a linear driving force (LDF) approximation. Simulation results indicate that, under certain conditions, equilibrium is not actually reached at the adsorbent surface. This leads to different unit performances, in terms of product purities and recoveries, as compared to those predicted assuming instantaneous equilibrium. Moreover, SMB units may be improperly designed by the usual methods (flow-rate ratio separation regions) if non-equilibrium effects are overlooked. PMID- 10674942 TI - Enantiomer separation of alpha-ionone using gas chromatography with cyclodextrin derivatives as chiral stationary phases. AB - The gas chromatographic enantiomer separation of alpha-ionone was studied with three different chiral stationary phases using as chiral selectors: (1) heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin, dissolved in polysiloxane PS-086, (2) octakis(2,6-di-O-pentyl-3-O-trifluoroacetyl)-gamma-cyclodextrin and (3) octakis(2,6-di-O-pentyl-3-O-butanoyl)-gamma-cyclodextrin, both dissolved in polysiloxane SE-54. The influence of the concentration of the chiral selector in the polysiloxane, coated on Chromosorb P AW-DMCS 80-100 mesh, is described and discussed, as well as the effect of Chromosorb loading. The feasibility of the preparative gas chromatographic separation of the enantiomers of alpha-ionone is considered; in order to provide a term of comparison, the estimated performances are compared with those achieved in the separation of the enantiomers of the inhalation anaesthetic enflurane. PMID- 10674943 TI - Preparative chromatographic resolution of enantiomers using polar organic solvents with polysaccharide chiral stationary phases. AB - The preparative chromatographic resolution of racemic mixtures is rapidly becoming a standard approach for the generation of enantiomers in pharmaceutical research and development. This paper will discuss the optical resolution of numerous pharmaceutical intermediates and final products using polar organic solvents with polysaccharide chiral stationary phases. The advantages of this approach compared to more traditional mobile phases for preparative separations will be presented. In addition the ability to reverse elution order using polar organic solvents will be presented. PMID- 10674944 TI - Direct, preparative enantioselective chromatography of propranolol hydrochloride and thioridazine hydrochloride using carbon dioxide-based mobile phases. AB - In this paper, we describe the direct, preparative enantioselective chromatography of racemic (rac)-propranolol hydrochloride (HCI) and rac thioridazine.HCl using Chiralpak AD chiral stationary phase and mobile phase systems containing carbon dioxide and methanol without the use of basic or acidic additives. Isolated fractions of propranolol.HCl were positively identified by mass spectrometry, Beilstein flame test, melting point, and chemical analysis to be HCI enantiomers of propranolol-HCl salts exhibited characteristic mass spectra peaks at 36 and 38 mass-to-charge ratio in the expected 3:1 isotopic ratio for the solute that were absent in the mass spectra for the free-base forms. To our knowledge, the direct, preparative enantioselective isolation of HCI enantiomeric salts of rac-propranolol and of rac-thioridazine have not been previously demonstrated and published. PMID- 10674945 TI - Epidemiological research in mental disorders: lessons for the next decade of research--the NAPE Lecture 1999. Nordic Association for Psychiatric Epidemiology. AB - OBJECTIVE: To provide an overview of the progress in the field of epidemiology of mental disorders during the last decade, and to discuss challenges for the future. METHOD: Following a review of the achievements of psychiatric epidemiology, current shortcomings in the research and publications in the field are discussed. Suggestions for changes in research and solutions to current deficits are proposed. RESULTS: Future research needs to cover a broader spectrum of psychopathology, including inquiries into sub-threshold characterizations; explore etiopathogenic processes and pathways including genetic, neuroscientific and psychological factors for the onset and persistence of psychopathology; and provide a better assessment of help-seeking behaviors and service utilization. CONCLUSION: Future epidemiological research should strengthen the emerging collaborative links with the neurosciences, clinical research and public health by acknowledging the full range of available epidemiological designs and methods, beyond the current emphasis on large-scale, cross-sectional general population studies. PMID- 10674946 TI - PET measurements of brain glucose metabolism and blood flow in major depressive disorder: a critical review. AB - OBJECTIVE: To show that PET investigations of brain function in patients with major depression can contribute with valuable pathophysiological knowledge about brain function of these states. METHODS: PET studies of cerebral blood flow or glucose metabolism in patients with unipolar or bipolar depression were reviewed. RESULTS: The studies have great discrepancies related to sample size, subject selection, imaging protocol and image analysis. In spite of this shortcoming, there is evidence that patients with major depression have reduced blood flow and metabolism in the prefrontal cortex, particularly when they exhibit psychomotor retardation. Abnormalities are also found in the anterior cingulate gyrus and the basal ganglia. A few studies point to the possibility that response to antidepressant treatment can be predicted from PET scans. CONCLUSION: This evidence is consistent with the hypothesis that depressive symptoms are caused by dysfunction of regions of the limbic system and the frontal lobes in close connection with the basal ganglia. PMID- 10674947 TI - Subtyping depression by clinical features: the Australasian database. AB - OBJECTIVE: To distinguish psychotic, melancholic and a residual non-melancholic class on the basis of clinical features alone. Previous studies at our Mood Disorders Unit (MDU) favour a hierarchical model, with the classes able to be distinguished by two specific clinical features, but any such intramural study risks rater bias and requires external replication. METHOD: This replication study involved 27 Australasian psychiatrist raters, thus extending the sample and raters beyond the MDU facility. They collected clinical feature data using a standardized assessment with precoded rating options. A psychotic depression (PD) class was derived by respecting DSM-IV decision rules while a cluster analysis distinguished melancholic (MEL) and non-melancholic classes. RESULTS: The MELs were distinguished virtually entirely by the presence of significant psychomotor disturbance (PMD), as rated by the observationally based CORE measure, with over representation on only three of an extensive set of 'endogeneity symptoms'. CONCLUSION: In comparison to PMD, endogeneity symptoms appear to be poor indicators of 'melancholic' type, confounding typology with severity. Results again support the hierarchical model. PMID- 10674948 TI - Generalized anxiety disorder (ICD-10) in primary care from a cross-cultural perspective: a valid diagnostic entity? AB - OBJECTIVE: Generalized anxiety disorder (GAD) is a common psychiatric disorder. The nosological status of this diagnostic entity was critically discussed because of the very high rate of comorbidity with other psychiatric disorders, the assumed low degree of social disability associated with GAD in the absence of other disorders, and an ambigious definition. METHOD: We explored the frequency and associated social disability of GAD, and examined whether the ICD-10 definition of GAD is appropriate. The analysis was based on the WHO study on 'Psychological Problems in Primary Care' conducted in a standardized manner in 14 countries. RESULTS: We found GAD (total and without another psychiatric disorder) to be common in primary care in nearly all countries (mean 1-month prevalence rate, 7.9%), with about 25% of these cases presenting with GAD in the absence of any comorbid psychiatric disorder. GAD in general, as well as non-comorbid GAD, are associated with social disability which is as severe as that in chronic somatic diseases. CONCLUSION: It remains questionable whether the current ICD-10 diagnosis of GAD defining 6 months as a minimum duration and requiring at least four associated symptoms for diagnosis is the most appropriate option. Using this definition, a substantial proportion of psychosocially disabled subjects characterized by anxiety, tension and worrying remain undetected, and are possibly therefore not adequately treated. PMID- 10674949 TI - Comorbidity of the anxiety disorders in a community-based older population in The Netherlands. AB - OBJECTIVE: The aim of the study was to investigate patterns of comorbidity among the anxiety disorders in a community-based older population, and the relationship of these disorders with major depression, use of alcohol and benzodiazepines, cognitive impairment and chronic somatic illnesses. METHOD: The data were derived from the Longitudinal Aging Study Amsterdam (LASA) study. A two-stage screening design was adopted to identify respondents with anxiety disorders. RESULTS: In total, 10% of the elderly with an anxiety diagnosis suffered from two or more anxiety disorders. Major depression (13% vs. 3%), benzodiazepine use (24% vs. 11%) and chronic somatic diseases (12% vs. 7%) were significantly more prevalent in respondents with an anxiety disorder than in respondents without anxiety disorders. Heavy or excessive alcohol intake (5% vs. 4%) and cognitive impairment (11% vs. 13%) were not significantly associated with any anxiety disorder. CONCLUSION: When anxiety disorders are diagnosed, in older people there is a relatively high probability of comorbid conditions being present. PMID- 10674950 TI - Traumatic events and post-traumatic stress disorder in the community: prevalence, risk factors and comorbidity. AB - OBJECTIVE: Lifetime and 12-month prevalence of traumatic events and DSM-IV post traumatic stress disorder as well as risk factors and comorbidity patterns were investigated in a representative community sample (n = 3021, aged 14-24 years). METHOD: Traumatic events and PTSD were assessed with the Munich Composite International Diagnostic Interview (CIDI). RESULTS: Although 26% of male subjects and 17.7% of female subjects reported at least one traumatic event, only a few qualified for a full PTSD diagnosis (1% of males and 2.2% of females). Traumatic events and PTSD were strongly associated with all other mental disorders examined. PTSD occurred as both a primary and a secondary disorder. CONCLUSION: The prevalence of PTSD in this young German sample is considerably lower than reported in previous US studies. However, the conditional probability for PTSD after experiencing traumas, risk factors and comorbidity patterns are quite similar. Traumatic events and full PTSD may increase the risk for other disorders, and vice versa. PMID- 10674951 TI - A transcultural outcome study of adolescent eating disorders. AB - OBJECTIVE: The aim of the study was to assess the treatment and outcome of adolescent eating disorders in an international study including Western and Eastern European clinical and research centres. METHOD: A total of 138 patients with adolescent onset of an eating disorder (primarily anorexia nervosa) were followed-up after a mean interval of 5 years after first admission. RESULTS: On average, the patients had spent 25% of the total follow-up period in either in patient or out-patient treatment. Half of them required a second hospitalization and a quarter required a third hospitalization for the eating disorder. At follow up, 68% of the total sample did not have an eating disorder. The prediction of outcome revealed different patterns of risk variables depending on the type of criterion. CONCLUSION: The outcome of adolescent eating disorders is relatively similar across cultures, and better than in patients with later onset of the disorder. PMID- 10674952 TI - Dangerous female psychiatric patients: prevalences and characteristics. AB - OBJECTIVE: We wanted to study the prevalence and characteristics of dangerousness among female psychiatric patients. METHOD: A national survey was performed in Norway, covering all psychiatric in- and out-patient units. RESULTS: There were 329 persons reported to have a psychiatric disorder and to satisfy our criteria for dangerousness, giving a total prevalence of 9.9/100000 adults. There were 54 women, giving a female prevalence of 3.1/100 000. When compared to a matched sample of the men, fewer women were out-patients or had had jail sentences. We found no sex differences with regard to frequencies of psychosis, mental retardation, personality disorders, or drug or alcohol abuse. The women had an increased frequency of suicide-related and self-injurious behaviours and previous commitment for arson. CONCLUSION: The prevalence of women with psychiatric disorders who are considered to be dangerous was 3.1/100 000. The dangerous men did not show higher frequencies than the women for psychopathology, drug abuse, behaviour or criminality. PMID- 10674953 TI - Sources of coercive behaviours in psychiatric admissions. AB - OBJECTIVE: Coercion during psychiatric admissions has been a topic of debate for many years. Although there has been considerable research on patients' perceptions of coercion, there has been no work on who places pressures on patients to be admitted. METHOD: This article integrates interview data from interviews with patients, admitting staff and family and friends to describe the pressures brought to bear on patients to be admitted. RESULTS: Health-care professionals appear to be the most important source of pressures on patients, and to have the most impact on patients' perceptions of coercion. However, there are differences in type of pressure, and the pressures used by family and friends appear to have the most longstanding impact. CONCLUSION: Legal and clinical efforts to reduce the level of coercive pressures on patients need to recognize the importance of mental-health professionals, including especially those who are not legally mandated to participate in the admission process. PMID- 10674954 TI - 'Everybody looks at my pubic bone'--a case report of an adolescent patient with body dysmorphic disorder. AB - OBJECTIVE: Body dysmorphic disorder (BDD) was described for the first time more than 100 years ago, but it is still unknown to many clinicians. Although the onset usually occurs during adolescence, BDD has received little attention in the adolescent psychiatric literature. METHOD: The case and treatment of a 16-year old female patient is described. RESULTS: The patient, suffering from the overvalued belief of a dislocated pubic bone, a comorbid mild depressive episode, BDD associated rituals and social avoidance, was treated successfully with a combination of exposure and response prevention and 125 mg/day of doxepine. CONCLUSION: If BDD is diagnosed early in the course and treated appropriately, it is possible to obtain a satisfying outcome. PMID- 10674955 TI - Dynamics of mucosal dimensions after root coverage with a bioresorbable membrane. AB - BACKGROUND: So far, the clinical effects of the placement of a resorbable membrane for guided tissue regeneration have not been studied in humans in great detail. The dynamics of the resorptive processes, in particular, appear to be rather speculative. In the present longitudinal study, specific alterations of the dimensions of the dentogingival mucosa were explored after surgical root coverage by using a bioresorbable membrane and a coronally-repositioned flap. METHODS: The study population consisted of 14 patients with a total of 31 predominantly shallow, Miller class I, II or III recessions. The thickness of the masticatory and lining mucosa before and after surgical intervention was measured with an ultrasonic device. RESULTS: Mean (+/-sd) recession depth and width were 2.85+/-1.29 and 4.46+/-1.14 mm, respectively. After 12 months, 51+/-29% of the recession depth (p<0.001) and 13+/-35% of its width (n.s.) were covered. Root coverage seems to be rather defect-type sensitive with best results obtained at canines with relatively shallow recessions. Mucosal thickness was considerably increased after surgery with a gradual decrease during the following 9 months. Thus, thickness of the marginal tissue rose from 0.82+/-0.27 mm to 1.49+/-0.54 mm 3 months after placement of the membrane (p<0.001). After 12 months, a mean thickness of 1.03+/-0.40 mm was observed (p<0.001). Even more pronounced alterations were noted for the alveolar lining mucosa with a threefold increase of thickness 3 months after surgery and a gradual decrease to about 1 mm after 12 months. CONCLUSIONS: The present results point to the considerable space making capacity of the bioresorbable membrane which probably allows for the ingrowth of a granulation tissue derived from the underlying structures. The gradual decline in mucosal thickness between months 6 and 9 after surgery may be paralleled by the maturation of the granulation tissue while complete resorption of the membrane had been accomplished. PMID- 10674956 TI - Stain, plaque and gingivitis reduction by combining chlorhexidine and peroxyborate. AB - BACKGROUND: Previous studies have shown that using an oxidising agent in addition to chlorhexidine reduces staining. AIM: The purpose of the present study was to investigate whether, compared to chlorhexidine alone, the use of an oxidising mouthrinse as an adjunct to chlorhexidine is efficacious in reducing stain, plaque and gingivitis. METHOD: This study had a single-blind, 2-group parallel design, including a 14-day experimental non-brushing period during which 1 group (n= 14) used chlorhexidine alone (CHX) (chlorhexidine mouthrinse, 0.12% Oral-B laboratories, Ireland), and the other (n= 14) used chlorhexidine in combination with an oxidising agent (sodiumperborate-monohydrate-Bocasan, Oral-B laboratories, Ireland). Patients were randomly assigned to either group. All participants received a scaling and polishing before the start of the trial. No oral hygiene instructions were given. Since, at the start of the experiment, all stain and plaque were removed, only the gingival condition was evaluated at baseline by means of bleeding on marginal probing. The examination after 14 days of rinsing included the evaluation of plaque, bleeding on marginal probing and stain (GMSI: gingival modification of the stain index). RESULTS: The results showed at day 14, a significant difference between the 2 groups for plaque (CHX: 0.18, CHX+PER: 0.08, p=0.03) and gingival bleeding (CHX: 0.38, CHX+PER: 0.21, p<0.001). The proportion of stained surfaces was less in the CHX+PER group (28%), than in the chlorhexidine group (48%) (p=0.04). CONCLUSIONS: In conclusion, the adjunctive use of an oxidising agent peroxyborate to chlorhexidine, proved to be superior to chlorhexidine alone with regard to the inhibition of plaque and development of gingivitis. In addition, the proportion of stained surfaces was significantly less when adding the oxidising mouthrinse to chlorhexidine. PMID- 10674957 TI - Evaluation of accuracy and variability of scoring-area-based plaque indices. A laboratory model. AB - BACKGROUND: Plaque scoring, using a variety of indices and methods, is widely used in clinical dentistry. There is limited information on inter- and intra examiner variability and almost no data on examiner accuracy. AIMS: The aim of this study, was to determine the inter- and intra-examiner variability and accuracy of 15 examiners, of differing plaque-scoring experience, in recording and judging plaque areas from simulated plaque on tooth charts. METHODS: Plaque, shaded red, was drawn onto tooth charts of 8 simulated "patients" and measured by planimetry by 1 investigator. For each tooth for each "patient", examiners subjectively copied the plaque onto blank charts and scored plaque in 5% increments for the global plaque index. This was repeated on 2 occasions. Drawn plaque areas were determine by the investigator and comparisons made with the actual areas and % scores of plaque. RESULTS: For both plaque recording methods, intra-examiner variability was low and slightly better than inter-examiner variability. Reproducing plaque areas showed a high level of accuracy in most examiners, as was judging areas in 5% increments, albeit slightly less accurate than area drawing. Overall examiner experience had little influence on judging plaque areas. CONCLUSION: The data suggest that area-based plaque indices can be scored or recorded accurately and with minimal variability within or between examiners. The laboratory model could be used to train and assess examiners. PMID- 10674958 TI - Explanatory models for clinical and subjective indicators of periodontal disease in an adult population. AB - BACKGROUND: The aim of this study was to analyze indicators of periodontal disease using: (1) community periodontal index of treatment needs (CPITN), (2) subjectively reported change of front teeth position, and 3) subjectively reported gingival bleeding. METHOD: These 3 indicators were used in models with explanatory variables from 4 domains: (A) socio-economic attributes, (B) general health and health-related lifestyle, (C) dental attitudes and behaviors, and (D) dental status expressed as (number of teeth and DFT) for the clinically determined dependent variables. In 1992, the study was carried out cross sectionally in all 50-year olds in 2 Swedish counties using a questionnaire (n=6343) and clinical investigation of a 20 % sub-sample (n= 1040). RESULTS: Multiple and logistic regression analysis showed that explanatory patterns varied for the clinical and subjective indicators. Use of tobacco had strong effects in all models as did high care utilization. There were few associations with socio economic attributes. The 2 subjective indicators "changed front position" and "gingival bleeding" associated with attitudes, behaviors and subjective health. Number of teeth and DFT covaried with clinical indicators. CONCLUSIONS: The main conclusions from this study are: (1) that it is possible to find multivariate models with acceptable goodness of fit for prediction of occurrence of periodontal indicators, and (2) that the lack of relation between social attributes and the disease gives arguments for a biological provenance of periodontitis. PMID- 10674959 TI - The effect of scaling and root planing on the clinical and microbiological parameters of periodontal diseases: 12-month results. AB - BACKGROUND/AIMS: Previously, we reported that SRP resulted in a decrease in mean pocket depth and attachment level and reduced prevalence and levels of Bacteroidesforsythus, Porphyromonas gingivalis, and Treponema denticola at 3 and 6 months post-SRP in 57 subjects with adult periodontitis. 32 of the 57 subjects were monitored at 9 and 12 months. Thus, the purpose of the present investigation was to evaluate the microbial and clinical effects of SRP in 32 (mean age 48+/ 11) subjects over a 12-month period. METHOD: Clinical assessments of plaque, gingival redness, suppuration, bleeding on probing, pocket depth and attachment level were made prior to SRP and at 3, 6, 9, and 12 months post-therapy. Subgingival plaque samples were taken at each visit and analyzed using the checkerboard DNA-DNA hybridization technique for the presence and levels of 40 subgingival species. Each subject also received maintenance scaling at each of the subsequent monitoring visits. Differences in clinical parameters and prevalence and levels of bacterial species were analyzed pre- and post-therapy using the Wilcoxon signed ranks test. The Quade test for related samples was used for analysis of multiple visits. RESULTS: Mean pocket depth (mm+/-SEM) decreased from 3.2+/-0.3 at baseline to 2.9+/-0.3 at 12 months (p<0.01). Mean attachment level showed significant reduction at 6 months, but did not diminish further. Bleeding on probing and plaque were significantly reduced at 12 months (p<0.001, p<0.05, respectively). P. gingivalis, B. forsythus and T. denticola decreased in prevalence and levels up to the 6-month visit and remained at these lower levels at 9 and 12 months. Significant increases in levels and prevalence were noted at 12 months for Actinomyces naeslundii genospecies 2, Actinomyces odontolyticus, Fusobacterium nucleatum ss polymorphum, Streptococcus mitis, Capnocytophaga sp, and Veillonella parvula. CONCLUSIONS: The data suggest that the maintenance phase of therapy may be essential in consolidating clinical and microbiological improvements achieved as a result of initial therapy. PMID- 10674960 TI - Comparative effects of quaternary ammonium mouthrinses on 4-day plaque regrowth. AB - BACKGROUND: Quaternary ammonium compounds constitute a large group of antibacterial chemicals with a potential for inhibiting plaque and gingivitis. One compound, benzalkonium chloride (BC), may be of value, although there is a dearth of evidence to support efficacy. The aim of this study was to measure the ability of 2 BC mouthrinses (0.05% and 0.1%) to inhibit de novo plaque reformation. METHOD: A 4-day plaque regrowth model. For comparative purposes, a commercial mouthrinse containing cetyl pyridinium chloride (CPC) and a positive control chlorhexidine (CX) mouthrinse were also evaluated. RESULTS: Compared to water control, a reduction in plaque scores of 52% was noted for the CX mouthrinse, 22.5% for CPC and 5% and 6% for the 2 BC rinses. For plaque area, reductions of 84%, 47%, 16% and 15% were found for CX, CPC, and the 2 BC rinses, respectively. Significant reductions in plaque area compared to the water rinse were also seen with the 2 BC rinses (p<0.05). However, for both plaque score and plaque area, the CX and CPC rinses significantly reduced plaque compared to the BC rinses (p<0.0001). CONCLUSIONS: These findings would suggest that the 2 benzalkonium rinses would only have a limited value at inhibiting plaque formation. PMID- 10674961 TI - Characterization of bone resorbing activity in gingival crevicular fluid from patients with periodontitis. AB - BACKGROUND: In attempts to elucidate factors stimulating bone resorption in patients with different inflammatory diseases in the vicinity of the skeleton, e.g., peridontal disease and rheumatoid arthritis, we are investigating the presence of bone-resorbing activity in a variety of inflammatory exudates. The aim of the present study was to characterize the bone-resorbing activity present in patients with periodontitis. METHODS: Bone-resorbing activity was assessed in gingival crevicular fluids (GCFs) collected from patients with periodontitis and from patients with no signs of gingivitis. Bone-resorbing activity was evaluated by analyzing the capacity of GCFs to stimulate the release of minerals and the breakdown of bone matrix proteins in cultured neonatal mouse calvariae. The concentrations of IL-1alpha, IL-1beta and PGE2 were determined with ELISA and RIA techniques, respectively. RESULTS: GCF eluates from 24 different healthy sites caused a 1.23+/-0.05 fold stimulation of 45Ca release, whereas GCF eluates from 45 different diseased (periodontitis) sites caused a 2.46+/-0.10 fold stimulation. The effect on 45Ca release was time- and concentration-dependent, inhibited by 3 different osteoclast inhibitors and associated with enhanced release of 3H from [3H]-proline-labelled bones. The activity in GCF causing enhanced 45Ca release was unaffected, or in some samples partially reduced, by ultrafiltration using a filter with a molecular weight cut-off of 3000 Daltons. The bone-resorbing activity was temperature sensitive (+90degrees C, 10 min). The concentrations of prostaglandin E2 (PGE2) in the diluted GCF eluates, used in the bone resorption bioassay, were too low to be responsible for the release of 45Ca. Antisera specifically neutralizing human IL-1a inhibited the stimulatory effect of GCF pooled from several diseased sites. The specific, recombinant human IL-1 receptor antagonist completely inhibited the effect of pooled GCFs. GCF eluates from diseased sites contained human IL-1alpha and IL-1beta at concentrations of 1838+/-294 pg/ml and 512+/-91 pg/ml, respectively. CONCLUSIONS: These data show that GCF contains activity(ies) stimulating osteoclastic bone resorption in vitro. The factor primarily responsible for this activity seems to be IL-1alpha, but IL-1alpha is not the sole activator of bone resorption in GCF. PMID- 10674962 TI - Concentration of 3 tetracyclines in plasma, gingival crevice fluid and saliva. AB - BACKGROUND: Systemically-administered tetracyclines have been used widely for treatment of periodontal diseases with little understanding of their delivery characteristics to periodontal tissues. This study was designed to measure concentrations of 3 tetracyclines in gingival crevice fluid (GCF), plasma and saliva of following systemic administration. METHOD: The concentration of tetracycline (TC), minocycline (MN) and doxycycline (DX) was measured in gingival crevice fluid (GCF), plasma and saliva of 20 subjects following single sequential standard oral systemic doses. Gingival crevice fluid concentration was measured at 4 sites (2 shallow and 2 deep) before administration, and at 1 h and 2 h following administration. Plasma and saliva concentrations were measured from in samples at the same time points. No antibacterial activity was detected before administration. The highest concentrations were measured 2 h after administration. RESULTS: The average concentrations at 2 h were highest in plasma (TC = 1.02, MN=2.18, DX=2.35 microg/ml). Intermediate concentrations were measured in GCF (TC=0.61, MN= 1.49, DX= 1.65 microg/ml). Saliva concentrations (TC=0.09 MN=0.31, DX=0.47 microg/ml) were the lowest of the 3 fluids monitored. Data are presented indicating that the average GCF concentration of systemically administered tetracyclines is less than the that of plasma concentration. The concentration of tetracyclines in GCF was strongly associated with plasma concentration, indicating a primary role of drug absorption in the delivery of these systemically administered antibiotics to the site of action in periodontal therapy. The average GCF concentration in individuals varied widely (between 0 and 8 microg/ml) with approximately 50% of samples not achieving levels of 1 microg/ml. CONCLUSION: These observations suggest that poor absorption of orally administered tetracyclines in many individuals may account for much of the variability in clinical response to antibiotics observed in practice. PMID- 10674963 TI - Exposure to tobacco smoking and periodontal health. AB - BACKGROUND: The influence of smoking behavior on the periodontal health condition was clinically and radiographically studied in 257 dentally aware adults in the age range 20-69 years, including 50 current smokers, 61 former smokers and 133 non-smokers. AIMS: The clinical variables to be investigated were frequency of diseased sites > or =4 mm, frequency of gingival bleeding sites and plaque index. In addition, the periodontal bone height was radiographically assessed as a % of the dental root length. METHODS: All variables were based on full-mouth examinations including all teeth and periodontia. RESULTS: The observations indicated an inferior periodontal health condition associated with smoking. This was evidenced by a significantly greater frequency of diseased sites and a significantly greater reduction of periodontal bone height in current smokers as compared to non-smokers. The condition of former smokers was intermediate between current smokers and non-smokers, suggesting that former smokers who have quit smoking have a better periodontal health condition than current smokers, although worse than that of non-smokers. The finding that former smokers exhibited less disease than current smokers suggests that smoking cessation may be beneficial and mitigate the untoward effects inflicted by smoking, allowing a normalization towards non-smoker conditions. Heavy exposure was consistently associated with more severe a condition than light exposure, suggesting that the relationship between smoking exposure and periodontal morbidity is dose-dependent. CONCLUSIONS: Altogether, the present observations identify a negative impact from smoking on periodontal health and provide further evidence that tobacco smoking is an avoidable risk for periodontal disease. PMID- 10674964 TI - Leukocyte functions in 2 cases of Papillon-Lefevre syndrome. AB - AIM: To investigate the role of leukocytes in the pathogenesis of Papillon Lefevre syndrome (PLS). METHODS: Peripheral blood polymorphonuclear neutrophils (PMNs), monocytes (MNs) and gingival crevicular fluid (GCF) were obtained from 2 cases of PLS with typical features. The chemotaxis of PMNs and MNs were evaluated using a modified Boyden chamber. The adherence of PMNs was determined by adherence of PMNs to petri dishes. Interleukin-8 (IL-8) in GCF was detected by sandwich ELISA. Elastase activity in GCF was measured with a low molecular weight substrate (S-2484) specific for granulocyte elastase. RESULTS: PMNs from both patients showed depressed chemotactic response to FMLP and IL-8. Total amounts of IL-8 in GCF from the 2 patients were much higher than those of the normal controls. Elastase activity was not significantly different from that of the controls. The adherence of PMN and the chemotaxis of MN in the 2 patients were normal. CONCLUSION: The depressed chemotactic response of PMN leads to decreased recruitment of PMN and/or release of lysozyme from PMN in the diseased gingival tissue, increasing the susceptibility of PLS patients to periodontal infection. PMID- 10674965 TI - After-effects of stress on crevicular interleukin-1beta. AB - BACKGROUND: In a previous study, we found stress to increase crevicular interleukin-1beta (Il-1beta) secretion induced by supragingival plaque. While in that study, stress and plaque were presented concomitantly, we now wondered whether a consecutive presentation of these 2 factors would still exert stress effects. METHOD: 39 medical students participated in the study; 18 took part in a major exam while the remaining 21 served as controls. From the day after the last exam, students neglected oral hygiene in 2 antagonistic quadrants for 21 days (experimental gingivitis), while they maintained perfect hygiene at the remaining sites. Crevicular fluid samples were taken at days 0, 5, 8, 15, 18, and 21 of experimental gingivitis. RESULTS: A significant effect of pre-exposure to academic stress on crevicular Il-1beta concentration was found (area under the curve: p=0.042), the effect size, however, being smaller than in our previous study when stress and plaque were presented concomitantly. CONCLUSIONS: It is concluded that pre-exposure to stress may persistently alter the immunological effects of microbial challenge to the periodontium. PMID- 10674966 TI - Hardy-Weinberg testing for HLA class II (DRB1, DQA1, DQB1, and DPB1) loci in 26 human ethnic groups. AB - Testing the fit of population data to Hardy-Weinberg proportions is crucial in the validation of many current approaches in population genetic studies. In this paper, we tested fit to Hardy-Weinberg proportions using exact approaches for both the overall and individual heterozygote genotype data of four HLA Class II loci: DRB1, DQA1, DQB1, and DPB1, from 26 human populations. Eighty of 99 overall tests fit the Hardy-Weinberg expectation (73% for DRB1, 89% for DQA1, 81% for DQB1 and 81% for DPB1). Deviations from Hardy-Weinberg proportions were both locus and group specific. Although we could not rule out other mechanisms at work, the individual test results indicated that the departure was possibly partly due to recent admixture. Evidence for selection and other sources of deviation are also discussed. PMID- 10674967 TI - HLA associations in type 1 diabetes among patients not carrying high-risk DR3-DQ2 or DR4-DQ8 haplotypes. AB - Type 1 diabetes is a complex disease where numerous genes are involved in the pathogenesis. Genes that account for approximately 50% of the familial clustering of the disease are located within or in the vicinity of the HLA complex on chromosome 6. Some DRB1, DQA1 and DQB1 genes are known to be involved, in addition to as yet unidentified HLA-linked genes. The DR4-DQ8 and DR3-DQ2 haplotypes are known to confer high risk for developing the disease, particularly when occurring together. Approximately 10% of patients, however, do not carry any of these high-risk HLA class II haplotypes. We have performed genotyping of DRB1, DQA1 and DQB1 alleles in non-DR3-DQ2/non-DR4-DQ8 patients and controls from Sweden and Norway to test if any HLA associations were observed in these patients. Our results clearly demonstrate several statistically significant differences in the frequency of HLA haplotypes between patients and controls. Case-control analysis including the relative predispositional effect test, and transmission disequilibrium test (TDT) analysis in Norwegian type 1 diabetes families revealed that the DQA1*03-DQB1*0301, DQA1*0401-DQB1*0402, DQA1*0101 DQB1*0501, DQA1*03-DQB1*0303 and DQA1*0102-DQB1*0604 haplotypes may also confer risk. Our analyses also supported independent risks of certain DRB1 alleles. The study clearly demonstrates that HLA associations in type 1 diabetes extends far beyond the well-known associations with the DR4-DQ8 and DR3-DQ2 haplotypes. Our data suggest that there is a hierarchy of HLA class II haplotypes conferring risk to develop type 1 diabetes. PMID- 10674968 TI - DQCAR 113 and DQCAR 115 in combination with HLA-DRB1 alleles are significant markers of susceptibility to rheumatoid arthritis in the Korean population. AB - We have investigated HLA region microsatellite polymorphisms in rheumatoid arthritis (RA) which are known to be associated with HLA class II alleles in the Korean population. Ninety patients with RA and 106 controls were employed for this study, in which TAP1CA, DQCAR, D6S273, HLA-DRB1, -DQA1 and -DQB1 allele typing were performed. DQCAR 113 (RR = 3.2, P<0.0002), DQCAR 115 (RR = 3.6, P<0.0001) and heterozygous DQCAR 113/115 (RR = 11.2, P<0.0001) frequencies were significantly increased in the RA group compared with the control group. The HLA DRB1 genotypes of patients who had DQCAR 113/115 alleles were defined as DRB1*04 and/or DRB1*09. There was no significant difference between RA and controls in D6S273 and TAP1CA allele frequencies. We demonstrated that HLA-DRB1*0405 (RR = 6.6, P<10(-6)), DQA1*03 (RR = 5.2, P<10(-6)), DQB1*04 (RR = 3.5, P<0.002) alleles were useful markers of susceptibility to RA in Koreans. The frequency of HLA DRB1*0405 was higher in DQCAR 113 allele-positive RA (68.1%) than in DQCAR 113 allele-negative (16.3%) and total RA (43.3%) groups, and the susceptibility risk of DQCAR 113 allele to RA was more increased in the DRB1*0405-positive group (RR = 5.5, P<0.04). On the other hand, DQCAR 115 allele was more significantly associated with susceptibility to RA in HLA-DRB1*0405-negative patients (RR = 5.1, P<0.0005), and the association between RA and HLA-DRB1*0405 was also significantly associated with DQCAR 115 allele-negative patients (RR = 13.2, P<0.00001) as compared with DQCAR 115 allele-negative control groups. HLA DRB1*0405-DQA1*03-DQCAR113-DQB1*03 haplotype showed high relative risk value (RR= 17.7, P<0.0002). In conclusion, the DQCAR allele in combination with HLA class II, especially DR, is probably a useful risk marker for RA susceptibility in the Korean population. PMID- 10674969 TI - Trans-speciation maintenance in the MHC region of a polymorphism which includes a polymorphic dinucleotide locus, and the de novo arisal of a polymorphic tetranucleotide microsatellite. AB - Alleles and the surrounding regions of DQCAR, a dinucleotide repeat tightly linked to HLA-DQB1, were sequenced in a range of primate species including man. Three polymorphic regions can usefully be defined in the description of these sequences: the dinucleotide GT repeat itself, the anonymous region 5' of this repeat, and a variable CTGT repeat in the 3' region. The 5' sequence displayed six alleles in the individuals studied. One of these alleles was invariably associated with substitutions in the GT repeat and absence of the CTGT repeat, the others with pure, polymorphic GT repeats and variation in the numbers of CTGT repeats. Haplotypes can be classified by the allele in the 5' region. Those carrying allele 1 were only found in man, those with allele 2 in man, chimpanzee and gorilla. The third haplotype (indicated by the presence of allele 3) was found in chimpanzee, gorilla and orang-utan, the fourth in chimpanzee and gibbon, the fifth in baboon, guenon and mangabey and the sixth in guenon and macaque. The alleles in the 5' region, but from different species, are thus often more similar than alleles from the same species, a phenomenon already shown for some HLA genes. This suggests that major histocompatibility sequences and surrounding sequences shared a correlated evolutionary history. The new polymorphic tetranucleotide microsatellite (CTGT, 3rd region) has possibly arisen de novo from the pre-existing dinucleotide GT. This study provides information not only on the molecular evolution of this particular microsatellite but also of the trans-speciation maintenance of polymorphism of its surrounding sequences. PMID- 10674970 TI - Significant associations of HLA-B*5101 and B*5108, and lack of association of class II alleles with Behcet's disease in Italian patients. AB - Behcet's disease has been known to be strongly associated with human leukocyte antigen (HLA) B51, one of the split antigens of HLA-B5. An increased incidence of HLA-B51 in the patient group has also been reported in an Italian population. Since the B51 antigen has been recently identified to comprise nine alleles, B*5101-B*5109, we performed HLA-B51 allele genotyping by the polymerase chain reaction-sequencing based typing (PCR-SBT) method as well as serological HLA-A and -B typing among 21 Italian patients with Behcet's disease in order to investigate whether there is any correlation of one particular B51-associated allele with Behcet's disease. In addition, HLA class II genotyping was performed by the PCR-restriction fragment length polymorphism (RFLP) method. As a result, only the phenotype frequency of the B51 antigen was found to be significantly increased in the patient group as compared to the ethnically matched control group by the corrected P-value analysis (71.4% in patients vs. 17.9% in controls; chi2 = 14.26, Pc = 0.0042, R.R. = 11.5). In the B51 allele genotyping, 11 out of 15 B51-positive patients were B*5101 and the remaining four were B*5108, whereas all of 5 normal controls were B*5101, showing significant association of each allele with Behcet's disease. No significant difference was observed between the patient and control groups in the HLA class II allelic distribution. This study revealed a strong association of Behcet's disease in Italian with B*5108 as well as B*5101, providing important insight into the molecular mechanism underlying an HLA association with Behcet's disease. PMID- 10674971 TI - The CC-chemokine receptor 5 (CCR5) is a marker of, but not essential for the development of human Th1 cells. AB - The CC-chemokine receptor 5 (CCR5) has recently been described as a surface marker of human T cells producing type 1 (Th1) cytokines. Here we confirm that CCR5 is expressed on human Th1 but not on Th2 T-cell clones. Using intracellular cytokine staining, we show that alloantigen specific CD4+ T-cell lines derived from a CCR5-deficient individual (delta32 allele homozygote) contain high numbers of both interferon gamma (IFN-gamma) and interleukin (IL)-2 producing cells, low numbers of IL-10 producing cells and no IL4 or IL-5 producing cells when stimulated with phorbol ester and ionomycin in vitro. These results were similar to those obtained from alloantigen specific CD4+ T-cell lines derived from CCR5 expressing individuals. An enzyme-linked immunoabsorbent assay (ELISA) confirmed that the Th1 cytokine-positive cells from the CCR5-deficient individual were able to produce equal amounts of cytokines when compared to T-cell lines from CCR5 expressing individuals, These results demonstrate that CCR5-negative T cells display the same capacity of Th1 T-cell differentiation as T cells derived from CCR5-expressing individuals. Thus, CCR5 expression is not essential for differentiation of human Th1 T cells. PMID- 10674972 TI - Lack of a strong association of CTLA-4 exon 1 polymorphism with the susceptibility to rheumatoid arthritis and systemic lupus erythematosus in Japanese: an association study using a novel variation screening method. AB - CTLA-4 is considered to be one of the attractive candidates for the susceptibility genes to rheumatic diseases. In the present study, the association of CTLA-4 polymorphism with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) was examined in the Japanese population using the case control association analysis. Polymerase chain reaction-preferential homoduplex formation assay (PCR-PHFA) was applied for the screening of genetic variations and for the genotyping of a large number of samples. A greater proportion of Japanese patients with RA (44%) and SLE (44%) compared with healthy individuals (37%) had exon 1 49 G/G genotype, but the difference did not reach statistical significance. However, when the patients with RA and healthy individuals were stratified according to HLA-DRB1 alleles, a weakly significant increase of the positivity of CTLA-4 49G allele was observed in HLA-DRB1*0405-positive patients (87%) compared with DRB1*0405-positive healthy individuals (71%) (P = 0.014, odds ratio = 2.77). These results indicate that CTLA-4 exon 1 polymorphism does not contribute greatly to the susceptibility to RA and SLE, at least in Japanese, although the presence of CTLA4 49G allele could be a minor predisposing factor for RA in HLA-DRB1*0405-positive individuals. In addition, PCR-PHFA was shown to be useful for a mass screening of gene variations. PMID- 10674973 TI - Polymorphic analysis of the high-affinity tumor necrosis factor receptor 2. AB - The tumor necrosis factor receptor 2 (TNF-RII, CD120b, TNF-R p75/80) gene has recently been characterised. It is located on chromosome 1p362 and consists of 10 exons and 9 introns A number of biallelic polymorphisms have been found in exons 4, 6, 9 and 10 based on differences between published sequences. In this study we have used polymerase chain reaction methodology in association with sequence specific primers (PCR-SSP) incorporating mismatches at the 3' end to identify these polymorphisms. We were able to confirm the presence of a single biallelic polymorphism in exon 6 corresponding to a (T/G) at nucleotide 676 of TNF-RII mRNA (gb:M32315) which results in an amino acid change and three biallelic polymorphisms in exon 10 (in the3'UTR) corresponding to (A/G) at nucleotide 1663, (T/G) at nucleotide 1668 and a (C/T) at nucleotide 1690 of gb:M32315, whereas no polymorphisms were observed in exons 4 and 9. Here we report that in 192 unrelated UK Caucasian individuals the allele frequencies determined by direct counting were: 676-T (0.77), 1663-G (0.51), 1668-T (0.95), and 1690-T (0.64) and the calculated gene frequencies were; 676-T (0.52), 676-G (0.12); 1663-G (0.30), 1663-A (0.28); 1668-T (0.77), 1668-G (0.025); and 1690-T (0.40), 1690-C (0.20). Furthermore, the presence of an A allele at nucleotide position 1663 was found to be strongly associated with the presence of a C allele at nucleotide position 1690 and a G allele at nucleotide position 1668 whereas the presence of a G allele at position 1663 was associated with the absence of a C allele at nucleotide position 1690. PMID- 10674974 TI - Immunogenicity of P/Q-type calcium channel in small cell lung cancer: investigation of alpha1 subunit polyglutamine expansion. AB - The ectopic expression of neuronal P/Q-type voltage-gated calcium channels in small cell lung carcinoma (SCLC) is thought to induce antisynaptic autoimmunity in the paraneoplastic Lambert-Eaton myasthenic syndrome. The gene CACNL1A4, encoding the principal (alpha1A) subunit of this calcium channel, is mutated in several inherited neurological disorders. One of these disorders (spinocerebellar ataxia, type 6, or SCA-6) involves the expansion of a trinucleotide (CAG) repeat unit. We hypothesized that a somatic CAG repeat instability of this gene in neoplastic cells might generate a non-self epitope capable of initiating autoimmunity to P/Q-type calcium channels. We therefore analyzed the CACNL1A4 gene in SCLC lines established from metastases derived from seven individual patients (four associated with Lambert-Eaton myasthenic syndrome, one associated with myasthenia gravis, and two not associated with neurological autoimmunity). We compared their CAG repeat numbers (determined by polymerase chain reaction (PCR) amplification followed by separation of products on a 6% polyacrylamide/8M urea gel) to published norms and to DNA from a patient with SCA-6. The number of CAG repeats in SCLC DNA fell within a normal range whether or not the neoplasm was complicated by neurological autoimmunity. Therefore, it is unlikely that somatically unstable CAG repeat units in the gene encoding the P/Q-type voltage gated calcium channel account for this tumor protein's immunogenicity in the Lambert-Eaton myasthenic syndrome. PMID- 10674975 TI - In vitro exposure of acute promyelocytic leukemia cells to arsenic trioxide (As2O3) induces the solitary expression of CD66c (NCA-50/90), a member of the CEA family. AB - Arsenic trioxide (As2O3) is a useful drug for the treatment of acute promyelocytic leukemia (APL), acting through a complex mechanism involving the induction of apoptosis. We investigated by flow cytometry whether in vitro treatment of APL leukemic cells with As2O3 determined specific surface membrane changes. Twelve APL bone marrow aspirates were analyzed following 7 days of in vitro treatment with As2O3 (0.25, 0.5 and 2.5 microM) with regard to the expression of a series of differentiation antigens. Twelve acute myeloid leukemia (AML) samples of non-APL morphotype were analyzed as controls. Exposure of APL as well as non-APL samples to any concentration of As2O3 did not affect the expression of beta2 integrins (CD11a and CD11b), CD45 isoforms (RA, RB and R0), CD44/H-CAM, CD33 and the CEA-related antigen family members CD66ade and CD66b, thus failing to disclose any maturating effect. Of interest, in all APL samples (but not in AML) every tested dose of As2O3 determined a dramatic upregulation of CD66c display; intermediate concentration (0.5 microM) of As2O3 increased the median percentage of CD66c+ cells from 5% in control cultures (25th-75th percentile 2-12%) to 80% in drug-exposed cultures (25th-75th percentile 58-90%) (P<0.001). The induction of solitary expression of CD66c is a new finding which demonstrates As2O3 capability of generating phenotypic changes absolutely restricted to APL cells Moreover, these results provide experimental basis for considering the involvement of the newly described CD66 signalling pathway in As2O3-driven programmed cell death. PMID- 10674976 TI - High-throughput class I HLA genotyping using fluorescence resonance energy transfer (FRET) probes and sequence-specific primer-polymerase chain reaction (SSP-PCR). AB - We have developed a semi-automated HLA class I typing system utilising TET/TAMRA labelled fluorescence resonance energy transfer (FRET) hydrolysis probes. The results from 87 individuals are in full concordance with serology and conventional gel-based systems. This assay replaces labour-intensive conventional gel-based DNA typing and has a higher allelic resolution than serology. Our approach differs from previously published fluorogenic probe typing protocols in that it provides simultaneous typing of HLA-A, -B and -C loci to medium resolution. Furthermore, by using equipment that is not specific to FRET probe analysis our system has in-built expansion capacity to 384 reactions per plate, thus making it applicable to high-throughput population screening. Automation is achieved through the use of computer software which analyses direct input from the fluorescence reader, allowing high throughput with a low inherent error rate. PMID- 10674977 TI - Mhc-DQ-DRB haplotype analysis in the rhesus macaque: evidence for a number of different haplotypes displaying a low allelic polymorphism. AB - In the HLA-DRB subregion of man, five major groups of haplotypes, often displaying a remarkable polymorphism, are distinguishable. The polymorphism is thought to be generated by point mutation, microgene conversion and gene rearrangement by recombination. In order to gain insight into the organization of the rhesus macaque major histocompatibility complex (MHC) class II region, DRB genes from monkeys of different origins previously typed for their DQ genes were analyzed. At first DRB haplotypes were deduced from DQ-homozygous monkeys. The stability of these haplotypes was then examined in DQ-heterozygous monkeys by sequence-based typing for the presence of members of the DRB1*03 and DRB1*04 lineage, and for seven single alleles detected on the haplotypes. Six DRB haplotypes linked to the five most frequent and three haplotypes linked to less frequent DQ haplotypes were identified. Six novel DRB alleles were detected. The number of DRB genes per haplotype varied between two and four. The results altogether suggest that in rhesus macaques, in comparison to man, the DQ haplotypes are linked to only a small number of DRB haplotypes, the number and diversity of DRB haplotypes is larger, and the allelic polymorphism of a given haplotype is smaller. The diversity of the DRB haplotypes was partly due to the varying number and identity of genes linked to DRB1*03 and DRB1*04. Furthermore, the number of DRB1 genes themselves varied from zero to two. PMID- 10674978 TI - HLA-B*4021: hybrid linking the B15 and B40 families. AB - Twenty alleles encoding molecules with the B60 or B61 serologic specificity have been reported thus far. This study characterized a new allele encoding a molecule exhibiting partial serologic reactivity with B15- and B40-related alloantisera from unrelated Korean individuals. Based on the DNA sequence, it appears that the novel allele has a sequence identical to some of alleles in B*15 family including B*1501 in exon 2. The sequence in exon 3, however, is identical to alleles in the B*40 family (B*4001/07/10/12) and B*4803. This implies that the novel allele, B*4021, has evolved by a reciprocal gene recombination involving members of these two families. The haplotype associated with B*4021 is likely to be A11-Cw4-B*4021 DRB1*04-DRB4*01-DQA1*03-DQB1*0301 . PMID- 10674979 TI - Antibody responses to a mucosally delivered HIV-1 gp120-derived C4/V3 peptide. AB - T1-SP10MN(A) is a synthetic peptide containing a T-helper (Th), cytotoxic T cell (CTL) and a B-cell epitope derived from the HIV-1 gp120 envelope protein. This peptide can elicit both systemic and mucosal antibody responses following nasal immunization in various species. In the present study, three different mucosal immunization strategies were performed in rabbits to determine which induced a more vigorous antibody response to T1-SP10MN(A). Nasal immunization followed by nasal boosting was found to be superior at inducing both serum IgG and vaginal secretory IgA (S-IgA) when compared to nasal followed by vaginal boosting. Conversely, vaginal priming followed by vaginal boosting elicited minimal serum IgG and vaginal S-IgA responses to T1-SP10MN(A), but moderate levels of vaginal IgG were detected. This study further demonstrates that vaginal immune responses can be elicited by immunization at distant and local mucosal sites. PMID- 10674980 TI - LPS mediated production of IL-1, PGE2 and PGF2alpha from term decidua involves tumour necrosis factor and tumour necrosis factor receptor p55. AB - Prostaglandins, with cytokines involved as intermediate factors, may have an essential role in premature labour when infection is present. We therefore wanted to study tumour necrosis factor (TNF), in cytokine and prostaglandin production in reproductive tissue. Decidual cell cultures were established and cells were stimulated with lipopolysaccharides (LPS). Media concentrations of TNF, interleukin-1 (IL-1), IL-6 and prostaglandin E2 and F2alpha were analysed, and involvement of LPS receptor CD14, TNF and TNF receptors (p55 and p75) were analysed, by studying effects after administration of specific antibodies. LPS induced an early peak elevation of TNF, with a subsequent release of IL-1, IL-6 and prostaglandins. Antibodies against CD14 inhibited these LPS effects. TNF antibodies reduced production of IL-1 and prostaglandins, whereas no significant influence on IL-6 production was observed. Antibodies against the TNF receptor p55 reduced all observed TNF effects. In contrast, p75 antibodies did not influence cytokine or prostaglandin production in this system. Our results suggest that increased TNF production is a prerequisite for LPS stimulated production of IL-1 and prostaglandins from decidual cells. LPS may directly stimulate IL-6 production. Of the two TNF receptors studied, only p55 seemed to be involved in the TNF signal transduction. PMID- 10674981 TI - MUC1/episialin: a critical barrier in the female reproductive tract. AB - The female reproductive tract must resist microbial infections as well as support embryonic development, implantation and placentation. Reproductive tract mucins, in general, and Muc1/episialin, in particular, play key roles in implantation related events and in protection from microbial infection. High levels of mucin expression in the lower reproductive tract presumably affords protection against infection while down-regulation of uterine mucins has been suggested to provide access to the uterine surface. The present studies demonstrate that mucins, particularly Muc1, are effective barriers to embryo attachment. Furthermore, a strain of female Muc1 null mice in normal housing displays chronic infection and inflammation of the lower reproductive tract and markedly reduced fertility rates. This phenotype is not observed when Muc1 nulls are housed in a pathogen free environment indicating that this phenotype results from chronic microbial exposure. Only normal endogenous flora were isolated from the reproductive tracts of affected Muc1 null mice, suggesting that these bacterial species become opportunistic with loss of the mucin barrier. Staphylococcal adherence to lower reproductive tract epithelia was found to be mediated by cell surface mucin carbohydrates. Collectively, these studies demonstrate a critical barrier role for Muc1 in various aspects of female reproductive tract physiology. PMID- 10674982 TI - Antisperm antibodies: a critical evaluation and clinical guidelines. PMID- 10674983 TI - T cell transformation with Herpesvirus saimiri: a tool for neuroimmunological research. AB - The finite life span of human T lymphocytes and their requirement of regular restimulation frequently limit human T cell studies. Once infected with H. saimiri, however, human and monkey T cells are transformed to stable growth without the need for further restimulation. H. saimiri persists in human growth transformed T cells episomally and only a few viral genes are expressed. The release of infectious virus from transformed human T cells has not been observed. H. saimiri-transformed T cells have the phenotype of mature activated CD4+ or CD8+ T cells. Transformed T cells retain a structurally and functionally intact T cell receptor and respond specifically to recognition of their antigen. They produce Th1-like cytokines, provide B cell help, can be triggered to become cytotoxic, and are sensitive to a variety of apoptosis-inducing treatments. While H. saimiri-transformed T cells resemble native T cells in numerous aspects, their reactivity to CD2 is strikingly different: Native T cells are activated via CD2 by certain pairs of mAbs, but not by the mere binding of CD2 to its ligand CD58. In contrast, H. saimiri-transformed T cells are activated by a single crosslinked anti-CD2 mAb and also by interaction with CD58-bearing cells. PMID- 10674984 TI - Humoral immune responses to adenovirus vectors in the brain. AB - We have investigated the humoral immune response to E1-deleted adenovirus vectors encoding the lacZ gene introduced into the brains of mice. Injection of these non replicating vectors into the brain induced systemic antibody production to adenovirus vectors in dose dependent manner. Apparent antibody production to beta galactosidase, the product of the lacZ gene, was detected later than anti adenovirus antibody. Neutralizing antibody was not detected. This study demonstrates that E1-deleted adenovirus vectors injected into the brain trigger humoral immune responses to the adenovirus and its gene products, but they are not sufficient to block the infection of cells by adenovirus upon repeat injection. PMID- 10674985 TI - Bone marrow-derived dendritic cells pulsed with tumor homogenate induce immunity against syngeneic intracerebral glioma. AB - To evaluate the efficacy and toxicity of dendritic cell (DC) based therapy for intracerebral gliomas, we utilized a cell line derived from an astrocytoma that arose spontaneously in a VM/Dk mouse. This astrocytoma mirrors human gliomas phenotypically, morphologically and secretes transforming growth factor (TGF) betas, immunosuppressive cytokines secreted by human gliomas. Systemic vaccination of mice with DCs pulsed with tumor homogenate followed by intracranial tumor challenge produced a > 160% increase in median survival (p = 0.016) compared with mice vaccinated with PBS or unpulsed DCs (p = 0.083). Fifty percent of mice treated with pulsed DCs survived long-term. Immunologic memory was demonstrated by survival of mice rechallenged with tumor. Both cell-mediated and humoral immunity was induced. On histological examination only focal areas of demyelination at the tumor implantation site were present. There was no evidence that autoimmune encephalomyelitis was induced by DC vaccination. Therefore, in a murine model, vaccination with DCs pulsed with glioma tumor homogenate is a safe and effective therapy against a syngeneic glioma located in the immunologically privileged central nervous system (CNS). PMID- 10674986 TI - Tubby-like protein 1 as an autoantigen in cancer-associated retinopathy. AB - Cancer-associated retinopathy (CAR) is a rare paraneoplastic syndrome that is characterized by retinal degeneration. Two cDNA clones, recoverin and tubby-like protein 1 (TULP1), were isolated from a human retinal cDNA library by using serum from a CAR patient as the probe. Both recoverin and TULP1 are retina-specific protein, and TULP1 is a member of tubby gene family. A determination of the recognized amino acid sequence of TULP1 by the patient serum and immunohistochemical studies on the distribution of TULP1 in the retina were done in this study. PMID- 10674987 TI - Differential involvement of sympathetic nervous system and immune system in the modulation of TNF-alpha production by alpha2- and beta-adrenoceptors in mice. AB - In the present study, the regulation of tumor necrosis factor-alpha (TNF-alpha) production by alpha2- and beta-adrenoceptors located on noradrenergic nerve terminals and on macrophages was studied in endotoxaemic mice. We found that reduction of the sympathetic outflow by reserpine dramatically increased the lipopolysaccharide (LPS)-induced TNF-alpha production, demonstrating that the release of endogenous noradrenaline (NA), controlled by presynaptic alpha2 adrenoceptors, was a determinant factor in this model. By using alpha2- and beta adrenergic drugs (clonidine, CH-38083, isoproterenol, propranolol) we provided the first in vivo evidence that, beside the dominance of neuronal alpha2- and macrophage beta-adrenoceptors, the alpha2-adrenoceptors on macrophages were also involved in the modulation of LPS-induced TNF-alpha production. Since adrenergic drugs are widely used in the clinical practice, our findings may have therapeutical implications. PMID- 10674988 TI - Functional and molecular characterization of VIP receptor--effector system in rat developing immunocompetent cells: G protein involvement. AB - Changes in the functional characteristics for vasoactive intestinal peptide (VIP) receptor-effector system were evaluated in rat developing immunocompetent cells (from 1-week-old animals up to 12-week-old animals). These characteristics include [125I]VIP binding studies, cell cyclic AMP (cAMP) generation, analysis of [125I]VIP-receptor complexes by cross-linking experiments, as well as developed associated G proteins assayed by cholera and pertussis toxin-catalyzed ADP ribosylation and Western blot. The Scatchard analysis of binding data was consistent with the existence of two classes of VIP binding sites with K(d) values unaltered and B(max) increased during postnatal development. The efficiency of VIP stimulation of cAMP generation increased from 1-week-old rats to adult conditions. The VIP-receptor complex apparent molecular mass (52-55 kDa) remains unaltered, but it was significantly lower in 2-week-old than in 8-week old rats. ADP-ribosylated material by cholera toxin (CTx) was higher from 8-week old than from 2-week-old animals, while ADP-ribosylation by pertussis toxin (PTx) was quantitatively higher in 8-week-old rats. Results were confirmed when immunoblots for different G protein subunits were performed. PMID- 10674989 TI - Retrovirus mediated gene transfer of the self antigen MBP into the bone marrow of mice alters resistance to experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a prototypic model of organ specific autoimmunity. MHC class II restricted T-cells directed against myelin basic protein (MBP) have been shown to cause EAE in susceptible strains of mice. We have asked whether the introduction of a gene encoding an autoantigen (MBP) into the hematopoetic stem cells of mice would result in tolerance to that protein. We have introduced cDNA encoding the 21.5 kDa isoform of MBP into the hematopoetic stem cells of B10.PL (73NS), SJL, and B10 mice by retrovirus mediated gene transfer. Our experiments show expression of proviral MBP in peripheral blood and thymus following transplantation of genetically modified stem cells. Such expression does not result in deletion of MBP-specific T cells or tolerance to MBP, nor does it alter susceptibility to MBP-induced EAE in susceptible strains B10.PL and SJL. However, retrovirus-mediated gene transfer resulted in resistant B10 mice developing mild EAE. This report demonstrates that autoreactive MBP-specific T cells can be selected in the presence of endogenous antigen or an MBP-encoding retrovirus. PMID- 10674990 TI - Antibody directed against mannan of the Mycobacterium tuberculosis cell envelope provokes blood-brain barrier breakdown. AB - Previous studies demonstrated that blood-brain barrier (BBB) breakdown observed at the cerebral level during experimental allergic encephalomyelitis (EAE) arises from the presence of Mycobacterium tuberculosis in Freund's adjuvant and not only from the encephalitogenic antigens. The main objective of this study was to check if the mannan moiety of lipoarabinomannan present in the mycobacterial cell envelope is responsible for an immune response provoking a BBB breakdown. The results showed that: firstly, the complete Freund's adjuvant (CFA) contains a high release of polysaccharides; secondly, the rats immunized with the CFA present important serum concentration of anti-mannan antibody; and finally, a single 2-mg dose of anti-mannan antibody injected intravenously in naive rats provokes an immediate and reversible BBB breakdown. These results suggest that mannan arising from the solubilization of the mycobacterial cell wall in Freund's adjuvant induces a high production of anti-mannan antibody, which, in turn, provokes a BBB breakdown and possibly facilitates the induction of EAE. PMID- 10674991 TI - Partial synergy of bisindolylmaleimide with apoptotic stimulus in antigen specific T cells--implications for multiple sclerosis. AB - In multiple sclerosis (MS), induction of T cell apoptosis constitutes a promising therapeutic strategy. Recently, bisindolylmaleimide has been shown to be an effective treatment of experimental autoimmune encephalomyelitis, presumably due to enhancement of CD95-mediated T cell apoptosis. Therefore, we studied the effects of bisindolylmaleimide on human (auto)antigen-specific T cells. We observed a synergistic effect of bisindolylmaleimide with apoptotic stimulus assessed via caspase activity and annexin V-binding, but no potentiation of DNA fragmentation or cell death. Thus, bisindolylmaleimide might be useful for modulating T cell apoptosis, yet more potent substances have to be generated re establishing immunological control over auto-reactive T cells. PMID- 10674992 TI - An alkali-soluble factor present in normal brain tissue inhibits antigen-specific lymphocyte proliferation. AB - The brain has long been recognized as an immune privileged site and probably contains multiple immune regulatory factors. We have investigated the immune regulatory properties of brain tissue on cultured lymphocytes. Homogenized brain tissue inhibits proliferation to antigen, but stimulates proliferation in response to most mitogens. The inhibitory activity is destroyed by treatment with proteases or neuraminadase. The activity is in the insoluble fraction of the homogenate, but becomes soluble in 0.04 M NaOH. After gel filtration chromatography of the alkali soluble material, the suppressive activity is in the high molecular weight fraction which contains protein and carbohydrate. The brain homogenate blocks the effects of IL-2. The activity is not affected by neutralizing antibodies against regulatory cytokines, does not depend on Fas or FasL, and is not due to the presence of gangliosides. These data suggest that a brain glycoprotein or proteoglycan which is either membrane-bound or part of the extracellular matrix has immune regulatory effects in culture. The relevance of these findings to immune regulation in the intact animal deserves further investigation. PMID- 10674993 TI - Chronic stress in caregivers of dementia patients is associated with reduced lymphocyte sensitivity to glucocorticoids. AB - Caring for the chronically ill is associated with chronic distress. In view of the adverse effects of distress on cellular immune function, such distress may have implications for health. Indeed, it has been proposed that the hypothalamic pituitary-adrenal (HPA) axis is a potential psychobiological mediator of these effects. In this study, we observed that elderly caregivers experienced greater distress and increased salivary cortisol than non-caregivers. In addition, caregivers had blunted mitogen-induced lymphocyte proliferation, lower mitogen induced IL-2 production, and reduced lymphocyte sensitivity to glucocorticoids. These results indicate that chronic distress is associated with impaired cell mediated immunity which is, in turn, associated with elevated basal steroid levels and altered steroid immunoregulation at the level of the lymphocyte. PMID- 10674994 TI - Intrathecal release of sICAM-1 into CSF in neuroborreliosis--increased brain derived fraction. AB - In the present study, we report sICAM-1 concentration in the cerebrospinal fluid (CSF) and serum of patients with neuroborreliosis (NB, n = 11), compared to the data from a control group of patients with corresponding blood/CSF barrier dysfunction but without inflammation in the central nervous system (disc prolaps, DP, n = 11). In NB, the sICAM-1 concentration in CSF was increased up to six-fold (ranges: 6.6-42.8 ng/ml and 2.2-9.8 ng/ml for NB and DP respectively) with no change in serum sICAM-1. The corresponding sICAM-1 CSF/serum concentration quotients (Q(ICAM)) were in the ranges: 22.5-171.3 X 10(-3), and 8.8-27.8 X 10( 3) for NB and DP respectively. This finding can be explained by increase of the brain-derived fraction of sICAM-1 in NB. In one case we observed increased Q(ICAM) on 6th day after admission to the hospital (171.3 X 10(-3) at the time of the first lumbar puncture slightly increasing to 243.6 x 10(-3) five days later), followed by normalization, in two remaining repunctured patients we observed decreasing QICAM with normalizing Q(Alb). PMID- 10674995 TI - Increased plasma levels of interleukin-1, interleukin-6 and alpha-1 antichymotrypsin in patients with Alzheimer's disease: peripheral inflammation or signals from the brain? AB - Plasma concentrations of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), C reactive protein (CRP) and alpha-1-antichymotrypsin (ACT) in 145 patients with probable Alzheimer's disease (AD) and 51 non-demented controls were measured. To investigate the cellular activation of peripheral immune system, plasma levels of neopterin were also investigated. Plasma levels of IL-1 were detectable in 17 patients with AD (13%) and only in one control (2%) and average levels of IL-1 were higher in AD patients than in controls (p < 0.001). IL-6 plasma levels were detectable in a higher proportion of AD and controls (53% and 27%, respectively), and were increased in patients with AD (p < 0.001). Plasma levels of ACT were increased in patients with AD (p < 0.001) and CRP levels were in the normal range. Plasma levels of neopterin were slightly lower in AD patients than in controls, but differences were not statistically significant. No significant correlation was observed between IL-1 and IL-6 levels or neopterin and cytokine levels in plasma from AD patients. Plasma levels of ACT negatively correlated with cognitive performances, as assessed by the mini mental state examination (MMSE; R = -0.26, p < 0.02) and positively correlated with the global deterioration state (GDS) of AD patients (R = 0.30, p < 0.007). Present findings suggested that detectable levels of circulating cytokines and increased ACT might not be derived by activation of peripheral immune system of AD patients. Detection of these molecules might be used for monitoring the progression of brain inflammation associated with AD. PMID- 10674996 TI - Size-dependent retardation and resolution by electrophoresis of rigid, submicron sized particles, using buffered solutions in presence of polymers: a review of recent work from the authors' laboratory. AB - Capillary zone electrophoresis (CZE) was conducted in buffered solutions of polyacrylamide (PA) and polyethylene glycol (PEG) to find the degree and the manner in which separation and resolution of submicron-sized rigid spherical polystyrene sulfate and carboxylate particles were affected by the presence of those polymers. In resolving pairs of representative particles, maximal resolution was observed at or near the entanglement threshold concentration, c*, of the polymer. The value of that maximum represents a several-fold increase in resolution. Since c* can be calculated from intrinsic viscosity, and the latter from the molecular weight of the polymer (and some constants available in the literature), optimally resolving polymer conditions become predictable. The maximum can also be experimentally determined by measuring intrinsic viscosity and calculating c*, or by either systematically varying the concentration of a polymer of constant molecular weight or by varying the molecular weight of a polymer at constant concentration. An optimally resolving field strength is superimposed on the maximally resolving condition of polymer concentration and weight. PMID- 10674997 TI - Characterization of two types of mitochondrial creatine kinase isolated from normal human cardiac muscle and brain tissue. AB - Two types of mitochondrial creatine kinase (Mi-CK), sarcomeric (sMi-) and ubiquitous (uMi-)CKs, were isolated from normal human cardiac muscle and brain tissue, respectively, and their heterogeneity was characterized by means of isoelectric focusing (IEF). Octameric sMi-CK and uMi-CK were electrophoresed cathodic to cytoplasmic muscle-type creatine kinase isoenzyme (CK-MM) and dimeric Mi-CKs were found at the position of CK-MM on a cellulose acetate membrane. The electrophoretic mobilities of sMi-CK were similar to those of uMi-CK. Octameric sMi-CK was focused at pI 7.1-8.0 and dimeric forms at pI 6.55, 6.75, 6.85, and 6.95. New bands appearing at pI 6.65 and 6.75 after treatment of sMi-CK with carboxypeptidase B were found to be delysined forms. sMi-CK reacted with anti-sMi CK antibodies, and the immune complexes were focused at pI 5.8. The Km value of sMi-CK for creatine phosphate (PCr) was 1.19 +/-0.20 mmol/L (mean +/- standard error), the activation energy (Ea) was 108.3+/-1.2 kJ/ mol, and the residual enzyme activity after heating at 45 degrees C for 20 min was 79.6+/-1.9%. On the other hand, octameric uMi-CK was focused at pI 7.1-7.9 and the dimeric forms were focused at pI 6.6, 6.7, 6.8, 6.9, and 7.0. Delysined forms were focused around pI 6.3, 6.4, 6.8, and 6.9. uMi-CK reacted with anti-sMi-CK antibodies, and the immune complexes were focused at pI 5.8. The Km value of uMi-CK for PCr was 1.07+/-0.03 mmol/L, Ea of uMi-CK was 110.0+/-0.9 kJ/mol, and the residual enzyme activity after heating at 45 degrees C for 20 min was 90.3+/-0.4%. The sMi-CK and uMi-CK were hybridized and the hybrid Mi-CK appeared at pI 6.78, 6.98, and 7.1 7.95. The pIs of the hybrid Mi-CK were between those of sMi-CK and uMi-CK. As described above, sMi-CK and uMi-CK were slightly different from each other with respect to the pI and some enzyme characteristics. PMID- 10674998 TI - Reduction of tyrosine-phosphorylated proteins in involuted thymuses of stressed rats: a study using immunological methods. AB - In the present study, we investigated whether tyrosine phosphorylation was involved in thymic involution, which has been reported to correlate well with the effects of various kinds of stresses. Immunohistochemistry using the anti phosphotyrosine antibody showed that the immunoreactivity decreased remarkably in the involuted thymus of stressed rats as compared with the control thymus. Immunoblot analysis using the anti-phosphotyrosine antibody revealed the tyrosine phosphorylated proteins with apparent molecular masses of 120, 90, and 70 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were detected in the control thymus. The immunoreactive band corresponding to the three proteins decreased remarkably in the involuted thymus. Further, we found by immunoprecipitation experiments that the 120 kDa protein was p130cas, a crk associated src substrate. These findings suggest that tyrosine phosphorylation signaling may be involved in thymic involution. PMID- 10674999 TI - Analysis of galectin 1-mediated cell signaling by combined precipitation and electrophoresis techniques. AB - Galectin-1 (GAL1) is a beta-galactoside-binding protein that has been implicated in the regulation of viability of lymphoid cells. However, the signaling pathway governed by the binding of GAL1 to the cell membrane is not understood yet. As a first step toward the elucidation of GAL1-initiated signaling events, electrophoresis techniques such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and two-dimensional electrophoresis (2-DE) were used together with precipitation techniques. This allowed us to identify the membrane receptor of GAL1, and to characterize the signal resulting from the binding of GAL1 to this receptor. Our results demonstrate that the tyrosine phosphatase CD45 is the receptor for GAL1, and that the src-type tyrosine kinase Lyn is a target for the effects of GAL1/CD45 interactions in B-cells. Furthermore, these results show the usefulness of combined precipitation and electrophoresis techniques to analyze phosphotyrosine-dependent mechanisms during the study of cell functions. PMID- 10675000 TI - Specific gel electrophoresis method detects two isoforms of human intestinal alkaline phosphatase. AB - We have demonstrated that the 6.0% polyacrylamide disc gel electrophoresis (PAGE) method in the presence of 1% Triton X-100 clearly separated both normal molecular mass intestinal alkaline phosphatase (NIAP) and bone alkaline phosphatase (BAP) in serum regardless of the ABO blood group and the secretor status of the subjects. From the results under the usual 7.5% PAGE condition, overlapping mobilities of NIAP and BAP were found in particular in nonsecretor subjects after a high-fat meal. Under the above conditions, the apparent BAP percentage three hours after a meal was higher in nonsecretors than in subjects under fasting conditions, because NIAP activity in serum rose sharply following a high-fat meal. In contrast, under our 6.0% PAGE method, the NIAP and BAP were clearly separated from each other regardless of whether the subjects were fasting or had ingested a high-fat meal. In addition, an elevated level of the circulating NIAP can be another marker for patients with liver cirrhosis. Considering all these factors, the 6.0% PAGE method proposed by us is not only a useful method for the separation of intestinal alkaline phosphatase (IAP) isoforms, but can also be useful for the analysis of other usual AP isozymes. PMID- 10675001 TI - The contribution of fructose and nitric oxide to oxidative stress in hamster islet tumor (HIT) cells through the inactivation of glutathione peroxidase. AB - Reducing sugars, such as glucose and fructose, are known to play a role in the initiation of apoptosis in pancreatic beta-cells. The data collected in this study show that fructose increases H2O2 levels and lipid peroxidation of hamster islet tumor (HIT) cells, which originated from hamster pancreatic beta-cells. In an attempt to clarify the mechanism of this oxidative stress, we were able to show that glutathione peroxidase (GPx) is inactivated by fructose, and that mRNA expression of GPx is suppressed by fructose. Nitric oxide (NO) is also known to bring about apoptosis. The presence of NO increases intracellular peroxide levels in HIT cells as judged by flow cytometric analysis. These data suggest that fructose and nitric oxide suppress the activity or expression of GPx, and, as a result, permit an increase in intracellular peroxides or lipid peroxidation. This represents a major contribution to the main mechanism of apoptosis by fructose and NO. PMID- 10675002 TI - Two novel myogenic factors identified and isolated by sequential isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - A number of environmental factors were used experimentally to enhance myogenesis during muscle regeneration; however, many hormones and growth factors have been shown to have the ability to increase the rate of satellite cell division, but they only work on satellite cells that are already active in many animal experiments. Recently, the crushed muscle extract (CME) of rats was found to be able to trigger dormant adult rat satellite cells to re-enter the cell mytogenic cycle; however, the identity of the active factors present in rat CME remains unknown. In the present study, the CME was fractionated by the strategy of sequential isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with functional analysis by myoblast culture. Two satellite cell-specific myogenic factors were identified and purified from CME by this strategy. One of the factors has a molecular mass of around 7 kDa and another about 39 kDa. The factor of 39 kDa could be retained in heparin-Sepharose column and eluted with phosphate-buffered saline (PBS) containing 1 M NaCl, but the 7 kDa factor did not bind to the heparin column. These two purified myogenic factors could synergistically trigger the proliferation and differentiation of dormant satellite cells, whose progenies subsequently fuse in vitro, or fuse to pre-existing partially damaged muscle fibers to form full repair of the damaged muscle fibers or to form new myotubes to replace the completely damaged muscle fibers during the cascade of muscle healing and regeneration in vivo. The identities of these two myogenic factors are under study. PMID- 10675003 TI - Measurement of serum low density lipoprotein-cholesterol in patients with hypertriglycemia. AB - Low density lipoprotein-cholesterol (LDL-c) concentration measured by a homogeneous enzymatic assay was reported to correlate well with the modified beta quantification assay, especially in samples with high triglyceride (TG) concentration. In this study, we evaluated a homogeneous enzymatic assay, Cholestest-LDL assay system, in hypertriglycemic patient samples, and found that 56% (9/16) of serum samples with intermediate TG concentrations (2.27-4.52 mmol/L) showed more than 10% discrepancy with concentration by the modified beta quantification assay. Such serum samples originated from patients with hyperglycemia of type II a (three cases), type II b (two cases), type III (one case), and type IV (six cases). Differential staining of cholesterol and triglyceride after agarose gel electrophoresis revealed that these serum samples contained significant amounts of intermediate fractions between pre-beta- and beta-lipoproteins. Since lipoprotein (a), which migrates between pre-beta- and beta-lipoproteins, is not correlated with the discrepancy, we believe the intermediate fraction consists of intermediate density lipoprotein (IDL) and a chylomicron remnant. A part of IDL and chylomicron remnant, which contain a significant amount of triglyceride, might be measured as LDL-c by the homogeneous enzymatic assay, but not by the modified beta-quantification assay. PMID- 10675004 TI - Heat shock protein (hsp 70)-related epitopes are common allergenic determinants for barley and corn antigens. AB - IgE reactive components of barley and corn were compared using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with sera from workers exposed to complex bioaerosols. The antibody made against Arabdopsis heat shock protein (hsp 70) was used to identify those components equivalent to hsp 70 in molecular size. Components with a molecular mass of 69 kDa and 33 kDa were positively reacted, and immunoblots of two-dimensional polyacrylamide gel electrophoresis were compared. PMID- 10675005 TI - Covalent binding of polyethylene glycol to the surface of red blood cells as detected and followed up by cell electrophoresis and rheological methods. AB - Cyanuric chloride activated polyethylene glycol (PEG)-5000 was covalently coupled to murine and human red blood cells (pegylated RBC). Our purpose was to camouflage RBC receptors, which is necessary for parasite invasion, a process essential to sustain parasitemia. Cell electrophoretic mobility analysis (CEM) of pegylated RBC distinguished a new population of cells bearing characteristic CEM. Pegylation of RBC also modified their rheological properties, which were documented by evaluation of cell deformability (based on cell transit time through calibrated micropores) and cell aggregation (as measured by ultrasonic interferometry). Homologous transfusion of pegylated RBC into murine malaria infected mice had no significant effect on the cerebral malaria death rate in Plasmodium berghei-infected mice, but it reduced the peripheral blood parasitemia by a factor 2 while in Plasmodium yoelii infected mice, the parasitemia was dramatically reduced by a factor of 4. These experiments demonstrate that transfusion of pegylated RBC may inhibit peripheral parasitemia. Cell electrophoresis appears to be a useful tool to allow in vivo detection and to investigate the fate of transfused pegylated RBC. PMID- 10675006 TI - Blood compatibility of particulate carriers investigated by cell electrophoresis and particle electrophoresis: a preliminary report. AB - Polyethylene glycol (PEG) and dextran were covalently coupled, or only adsorbed, to the surface of three kinds of inorganic particles in order to shield their surface and reduce their nonspecific binding to red blood cells. Surface modification as well as interaction of particles with red blood cells was followed up by particle electrophoresis. This allowed a quick evaluation of the efficiency of polymer coupling. Moreover, the nonspecific binding of particles to red blood cells was easily investigated with cell electrophoresis, showing the inhibitory effect of immobilized PEG-5000 or dextran. The electrophoretic mobility analysis presented here may be used for screening blood compatibility of particulate drug carriers and could be helpful in formulating long-living circulating particles. PMID- 10675007 TI - Mammalian pituitary growth hormone: applications of free flow electrophoresis. AB - We have used free flow electrophoresis (FFE) technology to study the electrophoretic behavior of growth hormone (GH) molecules, GH secretory granules and GH cell subpopulations contained in pituitary glands of humans and rodents. GH activities in different electrophoresis fractions were measured by immunoassay or bioassay, viz., measurement of chondrocyte proliferation in the tibial growth plate of the hypophysectomized rat. Using FFE we discovered a peptide in human post mortem pituitary tissue and cryopoor human plasma that is active in the tibial line bioassay, is inactive in a GH immunoassay, and is neither GH nor a GH fragment. This peptide, called tibial peptide, has high anodal mobility and is readily separable from GH by FFE. Its molecular mass is approximately 5 kD. It is particularly rich in glycine. A partial amino acid sequence (residues 9-25) in the middle region of the peptide shows that 9 of the 16 residues are nonpolar. On the basis of results from other FFE experiments, using either GH-containing secretory granules or GH-producing cells, we believe that the peptide is stored within the secretion granule of a subpopulation of GH cells. On the basis of recent information elucidating the role of C peptide contained in the insulin storage granule of the pancreatic cell, we propose that the tibial peptide serves a similar role in the GH cell. Thus, not only may tibial peptide aid in proper alignment of disulfide bonds between GH monomers in the secretory granule, but, like the C peptide, it also appears to have biologic activity in its own right. PMID- 10675008 TI - Multistage electrophoresis system for the separation of cells, particles and solutes. AB - It is common to operate equilibrium-based separation methods, such as distillation and extraction, as multistage unit operations, in which equilibrium is presumably achieved within each stage. Two rate-based separation processes, free electrophoresis and magnetic particle separation, have now been operated in multistage mode. Preparative free electrophoresis of particles and solutes has resisted scale-up and is confined to a narrow range of ionic compositions. Natural convection induced in electrophoresis buffers by Ohmic heating has been a strong deterrent and has led to such measures as radial electrophoresis in Couette flow, free-flow electrophoresis, low-gravity electrophoresis, density gradient electrophoresis, and reorienting density gradient electrophoresis, to name a few. The short vertical electrophoresis path exploited in the last mentioned forms the basis for multistage electrophoresis. A thin-layer countercurrent distribution apparatus was designed and constructed so that up to 20 fractions could be collected on the basis of electrophoretic mobility by applying an electric field. The mixture to be separated starts in a bottom cavity, and successive top cavities collect fractions as separand particles or molecules are electrophoresed upward out of the bottom cavity. Mathematical models of this process were developed, and experiments were performed to verify the predictions of the models by collecting and counting particles in each cavity after fractionation. PMID- 10675009 TI - Application of binary buffer systems to free flow cell electrophoresis. AB - The applicability of free flow electrophoresis (FFE) was expanded towards processing of sensitive cells. The chamber medium was adjusted to a physiologic pH of 7.35 by a mixture of N-(2-hydroxyethyl)piperazine-N'-(3-propanesulfonic acid) (EPPS) and 2,2-bis(hydroxymethyl)-2,2'2"-nitrilotriethanol (BISTRIS). These substances proved to be nontoxic to sensitive cells such as human smooth muscle or thyroid cells. They enhanced the electrical conductivity of the medium only slightly so that a new cell electrophoresis separation medium could be prepared, which contained 30 mM NaCl together with or without 1 mM CaCl2 but did not generate problems of overheating the fluid. Suspended in this medium, human smooth muscle cells as well as human thyroid carcinoma cells remained viable single cells for at least 120 min. After this period they could be recultured to form monolayers. If electrophoresed in the Octopus preparative FFE device, they migrated as single cells and did not clot; therefore, their electrophoretic behavior could be determined exactly. PMID- 10675010 TI - The role of DNA hypermethylation in human neoplasia. AB - Cancer development and progression is dictated by a series of alterations in genes such as oncogenes, tumor suppressor genes, DNA repair genes, and others. DNA methylation is an epigenetic modification that is profoundly altered in most cancers. Recently, hypermethylation of CpG-rich areas located in the promoter of genes (CpG islands) has been shown to be commonly implicated in silencing tumor suppressor genes in cancer. By cloning and characterizing a large number of such CpG islands hypermethylated in colon cancer, we found that two processes explain most of these events. Age-related CpG island methylation in a subset of cells in normal tissues, followed by intensification of methylation in cancer cells explains the majority of hypermethylation events in colon cancer and may provide a mechanistic link between aging and cancer formation. Most of the other CpG islands methylated in colon cancer can be explained by a newly described phenotype, the CpG island methylator phenotype (CIMP) which results in multiple methylation events in a subset of cancers. CIMP accounts for the majority of sporadic colon cancers characterized by microsatellite instability, as well as most tumors with k-ras mutations. Understanding further the factors that lead to, and modulate, aberrant methylation in cancer may provide novel avenues for prevention and treatment of this disease. PMID- 10675011 TI - Application of electrophoresis technology to DNA analysis. AB - We used the variable number tandem repeat (VNTR) polymorphism and the ten short tandem repeat (STR) polymorphisms to study a number of disputed paternity cases in the Japanese population. For the determination of VNTR locus (D1S80) and the ten STR loci (vWA, F13B, TH01, TPOX, CSF1PO, F13A01, LPL, D3S1744, D12S1090, D18S849) we used polymerase chain reaction (PCR) amplification and the vertical polyacrylamide gel electrophoresis technique followed by SYBR green I staining. The irregular repeats were analyzed by sequencing from bands of vertical polyacrylamide gel electrophoresis using the latest gene analyzing equipment, the ABI PRISM 310 Genetic Analyzer. The probable genotypes of the deceased putative father were deduced by Komatu's method from the genotypes of the widow and the genotypes of their children. The calculation of paternity probability used the Essen-Moller formula and Bayes's theorem. Calculated in eleven loci, the distinguishing probabilities (DP) and the mean exclusion chance (MEC) were 0.9999 and 0.9989, respectively. Therefore, information obtained from eleven DNA polymorphisms is enough to determine paternity plausibility. PMID- 10675012 TI - Regulation of macrophage-specific gene expression by degenerated lipoproteins. AB - The effect of aggregated low-density lipoprotein (agLDL) on cell viability and macrophage-specific gene expression using human peripheral blood monocytes in culture was investigated. AgLDL suppressed activation-induced cell death of phorbol ester-treated macrophages. The inhibition of apoptosis was accompanied by downregulation of apoptosis-promoting proteases, including interleukin-1beta converting enzyme (ICE) and CPP32 and upregulation of anti-apoptotic cytokine (interleukin-1beta (IL-1beta)). In contrast, macrophage-colony stimulating factor (M-CSF) enhanced cell death of lipid-bearing macrophages, suggesting that the anti-atherogenic action of M-CSF is at least in part mediated through apoptotic elimination of macrophages. Then, we attempted to isolate the genes specifically induced by agLDL in macrophages using a subtraction-based cloning strategy. One of the genes isolated, termed LIG (LDL-inducible gene), encodes a human homolog of E2 ubiquitin-conjugating enzyme. Ubiquitination of multiple intracellular proteins was observed in agLDL-treated macrophages, which coincided with upregulation of LIG. These results suggest that LIG acts as a direct mediator of foam cell formation through polyubiquitination and subsequent degradation of cellular proteins with apoptosis-inducing properties. The regulation of apoptosis by macrophage-specific gene expression may contribute to foam cell formation and atherosclerosis. PMID- 10675013 TI - Short tandem repeat analysis in Japanese population. AB - Short tandem repeats (STRs), known as microsatellites, are one of the most informative genetic markers for characterizing biological materials. Because of the relatively small size of STR alleles (generally 100-350 nucleotides), amplification by polymerase chain reaction (PCR) is relatively easy, affording a high sensitivity of detection. In addition, STR loci can be amplified simultaneously in a multiplex PCR. Thus, substantial information can be obtained in a single analysis with the benefits of using less template DNA, reducing labor, and reducing the contamination. We investigated 14 STR loci in a Japanese population living in Sendai by three multiplex PCR kits, GenePrint PowerPlex 1.1 and 2.2. Fluorescent STR System (Promega, Madison, WI, USA) and AmpF/STR Profiler (Perkin-Elmer, Norwalk, CT, USA). Genomic DNA was extracted using sodium dodecyl sulfate (SDS) proteinase K or Chelex 100 treatment followed by the phenol/chloroform extraction. PCR was performed according to the manufacturer's protocols. Electrophoresis was carried out on an ABI 377 sequencer and the alleles were determined by GeneScan 2.0.2 software (Perkin-Elmer). In 14 STRs loci, statistical parameters indicated a relatively high rate, and no significant deviation from Hardy-Weinberg equilibrium was detected. We apply this STR system to paternity testing and forensic casework, e.g., personal identification in rape cases. This system is an effective tool in the forensic sciences to obtain information on individual identification. PMID- 10675014 TI - Phylogenetic analysis of picoplankton in Lake Biwa and application to legal medicine. AB - Three strains of picoplankton designated as brown, green, and pink belonging to the Synechococcus genus in cyanobacteria (approximately 1 microm in size) are found ubiquitously in Lake Biwa, Japan. However, they could not be morphologically discriminated from other bacteria such as Proteobacteria and Bacillus by microscopy. In this study, we attempted to use the polymerase chain reaction (PCR) analysis of the 16S ribosomal DNA (rDNA) from picoplankton for the diagnosis of death by drowning. A segment of 16S rDNA was sequenced in order to investigate their phylogenetic relationships and to design the specific primers. The PCR products from three picoplanktons were compared with those from five other cyanobacteria, Melosira (diatom), Staurastrum (green alga), bacteria from Lake Baikal, and humans. The picogram order of template DNA from picoplankton was specifically amplified by the primers. When the template of picoplankton was mixed with human tissue, at least 10 ng of template DNA was needed to obtain a PCR product. The efficiency of PCR was increased more than hundredfold by isolating the picoplankton from human lung tissue. The specific PCR products of the picoplankton were obtained from a formalin-fixed drowning body (lung and liver) that was found in a downstream river and Lake Biwa. The PCR analysis of the picoplanktion 16S rDNA is considered useful for the diagnosis of death by drowning. PMID- 10675015 TI - Large volume sample stacking of positively chargeable analytes in capillary zone electrophoresis without polarity switching: use of low reversed electroosmotic flow induced by a cationic surfactant at acidic pH. AB - A simple and effective way to improve detection sensitivity of positively chargeable analytes in capillary zone electrophoresis more than 100-fold is described. Cationic species were made to migrate toward the cathode even under reversed electroosmotic flow caused by a cationic surfactant by using a low pH run buffer. For the first time, with such a configuration, large volume sample stacking of cationic analytes is achieved without a polarity-switching step and loss of efficiency. Samples are prepared in water or aqueous acetonitrile. Aromatic amines and a variety of drugs were concentrated using background solutions containing phosphoric acid and cetyltrimethylammonium bromide. Qualitative and quantitative aspects are also investigated. PMID- 10675016 TI - Electropherogram of capillary zone electrophoresis with effective mobility axis as a transverse axis and its analytical utility. I. Transformation applying the hypothetical electroosmotic flow. AB - For capillary zone electrophoresis, a new method of transformation from migration time to effective mobility was proposed, in which the mobility increase due to Joule heating and the relaxation effect of the potential gradient were eliminated successfully. The precision of the mobility evaluated by the proposed transformation was discussed in relation to the analysis of rare earth ions. By using the transformation, almost the same pherograms could be obtained even from the pherograms obtained originally at different applied voltages. PMID- 10675017 TI - Automated DNA fragment collection by capillary array gel electrophoresis in search of differentially expressed genes. AB - An automatic DNA fragment collector using capillary array gel electrophoresis has been developed. A sheath flow technique is used for not only detection but also collection of DNA fragments. In a sheath flow cell, the DNA fragments separated by 16 capillaries flow independently into corresponding sampling capillaries. The fraction collector consists of 16 sampling trays and each sampling tray is set beneath each end of the sampling capillaries to collect the flow-through DNA fragments. Certain DNA fragments are automatically sorted by controlling the movement of the sampling trays according to the signals from the system. The collector experimentally separated two mixtures of polymerase chain reaction (PCR) products: one prepared by using eight different sizes (base lengths from 161 to 562) of DNAs; and the other prepared by a differential display (DD) method with cDNA fragments. Collected DNA fragments are amplified by PCR and measured by electrophoresis. DNA fragments with base length differences of one (base lengths 363 and 364) were successfully separated. A separated DNA fragment from the DD sample was also successfully sequenced. In addition, differentially expressed DNA fragments were automatically sorted by comparative analysis, in which two similar cDNA fragment groups, labeled by two different fluorophores, respectively, were analyzed in the same gel-filled capillary. These results show that the automatic DNA fragment collector is useful for gene hunting in research fields such as drug discovery and DNA diagnostics. PMID- 10675018 TI - Fluorescence-based single-strand conformation polymorphism analysis of mutations by capillary electrophoresis. AB - Capillary electrophoresis in combination with fluorescence-based single-strand conformation polymorphism (SSCP) analysis was used to screen for known mutations as well as for unknown mutations. The mutations causing hemochromatosis and thrombogenetic diseases (factor V Leiden mutation and prothrombin mutation) are well defined. Familial hypercholesterolemia is caused by mutations in the low density lipoprotein (LDL) receptor gene. Because the mutations are heterogeneously localized in all 18 exons of the LDL receptor gene, effective screening procedures are necessary. The three well known mutations and 59 of 61 previously characterized mutations in the LDL receptor gene were detected by a distinct abnormal fragment pattern in capillary electrophoresis. The remaining two mutations in the LDL receptor gene showed only slight abnormalities under standard electrophoresis conditions (13 kV, 30 degrees C, 30 min). However, the abnormal pattern could be amplified by increasing the electrophoresis temperature. In all cases, heterozygous and homozygous mutations could clearly be differentiated from wild-type alleles. Because of the high efficiency of mutation detection, capillary electrophoresis in combination with fluorescence-based SSCP analysis would be attractive for the detection of well-defined mutations as well as for the screening of unknown mutations. The accuracy and the degree of automation make this technique well suited for routine genetic diagnosis. PMID- 10675019 TI - Confirmation testing of amphetamines and designer drugs in human urine by capillary electrophoresis-ion trap mass spectrometry. AB - Monitoring of amphetamines and designer drugs in human urine is a timely topic in clinical toxicology, surveillance of drug substitution, forensic science, drug testing at the workplace, and doping control. Confirmation testing of urinary amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) and 3,4-methylenedioxyamphetamine (MDA) by capillary electrophoresis (CE) combined with atmospheric pressure electrospray ionization and ion trap mass spectrometry (MS) is described. Using an aqueous pH 4.6 buffer composed of ammonium acetate/acetic acid, CE-MS and CE-MS2 provided data that permitted the unambiguous confirmation of these drugs in external quality control urines. Furthermore, other drugs of abuse present in alkaline urinary extracts, including methadone and morphine, could also be monitored. The data presented illustrate that the sensitivity achieved with the benchtop MS is comparable to that observed by CE with UV absorption detection. CE-MS2 is further shown to be capable of identifying comigrating compounds, including the comigration of amphetamine with nicotine. PMID- 10675020 TI - Simultaneous determination of bromide, nitrite and nitrate ions in seawater by capillary zone electrophoresis using artificial seawater as the carrier solution. AB - We describe capillary zone electrophoresis (CZE) for the simultaneous determination of bromide, nitrite and nitrate ions in seawater. Artificial seawater was adopted as the carrier solution to eliminate the interference of high concentrations of salts in seawater. The artificial seawater was free from bromide ion to enable the determination of bromide ion in a sample solution. For the purpose of reversing the electroosmotic flow (EOF), 3 mM cetyltrimethylammonium chloride (CTAC) was added to the carrier solution. A 100 microm ID (inside diameter) capillary was used to extend the optical path length. The limits of detection (LODs) for bromide, nitrite, and nitrate ions were 0.46, 0.072, and 0.042 mg/L (as nitrogen), respectively. The LODs were obtained at a signal to noise ratio (S/N) of 3. The values of the relative standard deviation (RSD) of peak area for these ions were 1.1, 1.5, and 0.97%. The RSDs of migration time for these ions were 0.61, 0.69, and 0.66%. Artificial seawater samples containing various concentrations of bromide, nitrite, and nitrate ions were analyzed by the method. The error was less than +/-12% even if the concentration ratio of bromide ion to nitrite or nitrate ion was 20-240. The proposed method was applied to the determination of bromide, nitrite, and nitrate ions in seawater samples taken from the surface and the seabed. These ions in other environmental waters such as river water and rainwater samples were also determined by ion chromatography (IC) as well as this method. PMID- 10675021 TI - Capillary zone electrophoresis of albumin-depleted human serum using a linear polyacrylamide-coated capillary: separation of serum alpha-and beta-globulins into individual components. AB - The separation of human serum globulins into individual components was investigated by capillary zone electrophoresis (CZE) using a linear polyacrylamide-coated capillary at pH 7.4. Prior to CZE analysis of globulin components present in serum, it was found that it was necessary to remove albumin. Preparation of albumin-depleted human serum with a HiTrap Blue column allowed the detection of alpha- and beta-globulin components as a series of peaks. Almost all the peaks, both narrow and broad, observed in CZE analysis could be assigned to six globulin components (alpha1-acid-glycoprotein, alpha1 antitrypsin, haptoglobin, alpha2-macroglobulin, Gc-globulin, and transferrin) by using the technique of antibody-based indirect detection. The CZE results, obtained from serum preparations from three healthy adults and six patients, showed that the CZE system might be capable of detecting qualitative differences among individuals with regard to individual globulin components. PMID- 10675022 TI - Capillary electrophoresis for determination of free and albumin-bound bilirubin and the investigation of drug interaction with bilirubin-bound albumin. AB - Capillary electrophoresis (CE) is a promising technique for assessment of free bilirubin and its interaction with albumin, as it requires only a small sample volume and provides a rapid and efficient separation of free bilirubin from its albumin-bound complex in a one-phase system. In order to maintain the equilibrium without dissociation of bilirubin from the albumin/bilirubin complex as in real clinical conditions, the coupling of CE with frontal analysis (FA) was investigated. A very large sample plug was introduced hydrodynamically into the capillary (36 cm length, 50 microm inner diameter) at 15 psi x s to develop the frontal conditions during CE separation. The working conditions for CE/FA separation of bilirubin and albumin were optimized as follows: +20 kV; running buffer, 10 mmol/L phosphate and 1 mmol/L ethylenediaminetetraacetic acid (EDTA), pH 7.4. The working range for bilirubin was found to vary from 5 to 206 micromol/L; precision with relative standard deviation (RSD) <2.0% for n = 3 and detection limit (signal to noise, S/N = 2) was 2 micromol/L. The residual binding capacity of a simulated cord blood serum for bilirubin was 26 mg/100 mL at pH 7.4. Bilirubin was shown to be displaced from albumin when aspirin was added. The free bilirubin concentration was found to increase to clinical significant concentrations from 11.9 to 21.1% when increasing aspirin was added in the range of 5-50 mg/100 mL, respectively. Thus, the investigation of aspirin displacement of bilirubin from albumin is clinically important and the CE/ FA method is a suitable procedure for this purpose. PMID- 10675023 TI - Two-dimensional map of the proteome of Haemophilus influenzae. AB - We have constructed a two-dimensional database of the proteome of Haemophilus influenzae, a bacterium of medical interest of which the complete genome, comprising about 1742 open reading frames, has been sequenced. The soluble protein fraction of the microorganism was analyzed by two-dimensional electrophoresis, using immobilized pH gradient strips of various pH regions, gels with different acrylamide concentrations and buffers with different trailing ions. In order to visualize low-copy-number gene products, we employed a series of protein extraction and sample application approaches and several chromatographic steps, including heparin chromatography, chromatofocusing and hydrophobic interaction chromatography. We have also analyzed the cell envelope bound protein fraction using either immobilized pH gradient strips or a two detergent system with a cationic detergent in the first and an anionic detergent in the second-dimensional separation. Different proteins (502) were identified by matrix-assisted laser desorption/ionization mass spectrometry and amino acid composition analysis. This is at present one of the largest two-dimensional proteome databases. PMID- 10675024 TI - Mass spectrometric approaches for the characterization of proteins on a hybrid quadrupole time-of-flight (Q-TOF) mass spectrometer. AB - This study demonstrates structural and conformational characterization of proteins by nanoflow electrospray ionization (nanoESI) mass spectrometry (MS) and tandem mass spectrometry (MS/MS) utilizing a quadrupole time-of-flight (Q-TOF) mass spectrometer (Micromass, Manchester, England). Model peptides were successfully sequenced at the 35 attomole (amol) level, and peptides derived from a tryptic in-gel digest of 25 femtomole (fmol) bovine serum albumin (BSA) were successfully sequenced. The results demonstrated that the MS/MS sensitivity of the Q-TOF clearly surpassed the detection limit of the silver stain. A silver destaining step greatly improved the mass analysis of peptides derived from in gel digests. Interestingly, sequence analysis revealed BSA residue 424 (tyrosine) as a potential chlorination site. In addition, a modified procedure was successfully used to extract and measure the masses of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-resolved proteins in the 10-68.5 kDa range. The Q-TOF was also used to monitor conformational changes of proteins. These experiments demonstrated an acid-induced denaturation of BSA in the pH 3-4 range, and heat-induced unfolding of cytochrome c between 50 and 60 degrees C. Finally, Zn2+ binding was demonstrated for the carbonic anhydrase apoprotein. In summary, the wide range of applications and the high quality of the experimental data made the Q-TOF mass spectrometer a powerful analytical tool for protein characterization. PMID- 10675025 TI - Two-dimensional gel electrophoresis using immobilized pH gradient tube gels. AB - An apparatus for the preparation of gels for immobilized pH gradient isoelectric focusing (IPG) in glass tubes was developed. Using this apparatus, the highly reproducible immobilized pH gradient can be formed with Immobilines in polyacrylamide gels, and IPG gels at all possible pH ranges can be easily prepared at low cost. The IPG tube gels in the first dimension in two-dimensional gel electrophoresis was used to separate and identify a number of rice embryo proteins in the proteome analysis. There was no difference in resolution of proteins between the tube gels and the commercially available slab gels; after electrophoresis, however, we could efficiently obtain a larger amount of the purified proteins from the tube gels than from the slab gels. PMID- 10675026 TI - Agarose isoelectric focusing for the detection of many isoforms and high molecules in muscle protein analysis. AB - Two-dimensional gel electrophoresis with agarose in the first dimension was established. The technique was reported previously and was now much improved without changing the basic procedure, presenting "a protein map" of chicken skeletal muscle, including almost all components such as myosin heavy chains (200 kDa) and dystrofin (400 kDa). Application of this technique for the analysis of all components was also successful with the liver and lens. Since large molecules can get into the agarose in the first dimension, the study on protein interaction was also reported with troponin components as a model system. PMID- 10675027 TI - Involvement of protein kinase C epsilon in thyrotropin-releasing hormone stimulated phosphorylation of the myristoylated alanine-rich C kinase substrate in rat pituitary clonal cells. AB - We have shown previously that novel protein kinase Cepsilon (nPKCepsilon) plays a key role in the basal and thyrotropin-releasing hormone (TRH)-stimulated prolactin (PRL) secretion in rat pituitary GH4C1 cells (Akita et al., J. Biol. Chem. 1994, 269, 4653-4660). Here we examined the region downstream of nPKCepsilon activation in order to understand the molecular mechanism by which nPKCepsilon mediates TRH-induced signal transduction. Exposure of GH4C1 cells to TRH causes a stimulation of the phosphorylation of a p80 (Mr approximately 80 000, pI approximately 4.3) and two p19 (p19a and b; Mr approximately 19 000, pI approximately 5.6 and 5.5, respectively). Phorbol ester, a potent activator of protein kinase C (PKC), also enhances these phosphorylations, whereas bisindolylmaleimide I, a specific inhibitor of PKC, clearly inhibits the phosphorylation of p80. p80 and p19 were identified as myristoylated alanine-rich C kinase substrate (MARCKS) and stathmin, respectively, as assessed by their two dimensional gel electrophoretic profiles and their stabilities to heat and acid treatment. In nPKCepsilon-overexpressing stable clones, the phosphorylated level of MARCKS but not stathmin was high in the resting state, and enhanced and sustained upon TRH stimulation, correlating with the increased activation of nPKCepsilon. TRH stimulates the release of MARCKS from the membrane/cytoskeletal fraction to the cytosol fraction. These results, taken together with previous data concerning PRL secretion, suggest that MARCKS, a regulatory component of the cytoskeletal architecture, is the major substrate of nPKCepsilon in vivo, and that its phosphorylation may regulate TRH-stimulated PRL secretion. PMID- 10675028 TI - Proteomic approach to the identification of cell membrane proteins. AB - The expression of plasma membrane proteins in human monocyte-derived U937 cells was examined by cell disruption and isolation of microsomal fractions. Two alternative procedures for cell disruption, Dounce homogenization and nitrogen cavitation, were compared. Cell homogenization and sequential centrifugation resulted in an approximately fivefold enrichment of plasma membrane proteins in the microsomal fraction. However, identification of 30 such apparently enriched proteins by two-dimensional (2-D) electrophoresis, proteolytic digestion, and mass spectrometry revealed that only eight were plasma membrane proteins, the remaining 22 being contaminants. In contrast, nitrogen cavitation followed by sequential centrifugation and solubilization of proteins with sodium dodecyl sulfate (SDS) and 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS) detergent yielded subcellular fractions, including microsomes, that showed little overlap in constituent proteins as revealed by 2-D electrophoresis. These results highlight the importance of obtaining pure plasma membranes and complete solubilization of membrane proteins for proteomic analysis. PMID- 10675029 TI - Age-related changes of aqueous protein profiles in rat fast and slow twitch skeletal muscles. AB - Two-dimensional electrophoresis was used to generate the aqueous protein expression patterns of rat extensor digitorum longus muscle (EDL, fast twitch muscle) and solues muscle (SOL, slow twitch muscle) of different ages. Two specific protein spots, S1 and S3, were identified from EDL muscles at the ages of 12 and 18 months onward respectively. In the EDL muscles of aged rat (24 months) after intensive exercise training, S3 was still detected while S1 disappeared. In addition, diaphragm muscle (DIA, fast twitch muscle), which retains physically active throughout the life span, was used as nondisuse control. The results showed that the expressions of S1 and S3 in 24-month DIA muscle were identical with the trained aged EDL muscle. It is suggested that exercise might delay the onset of S1 expression. However, the expression of S3 over age seemed to be progressive and exercise independent. Another protein spot, S2 was identified to express only in young EDL and SOL muscles, but its expression decreased over age. Furthermore, exercise has no effect on S2 expression since S2 could not be detected in aged DIA as well as trained aged EDL and SOL muscles. These results indicated that aqueous protein expression patterns of skeletal muscle undergo changes during aging. Some of these changes such as S2 and S3 appear progressively, and some such as S1 could be delayed by exercise. S3 was identified as ubiquitin, which might play an important role in protein degradation during skeletal muscle aging process. PMID- 10675030 TI - Damage recognition in nucleotide excision repair of DNA. AB - Nucleotide excision repair (NER) is found throughout nature, in eubacteria, eukaryotes and archaea. In human cells it is the main pathway for the removal of damage caused by UV light, but it also acts on a wide variety of other bulky helix-distorting lesions caused by chemical mutagens. An ongoing challenge is to understand how a site of DNA damage is located during NER and distinguished from non-damaged sites. This article reviews information on damage recognition in mammalian cells and the bacterium Escherichia coli. In mammalian cells the XPC hHR23B, XPA, RPA and TFIIH factors may all have a role in damage recognition. XPC hHR23B has the strongest affinity for damaged DNA in some assays, as does the similar budding yeast complex Rad4-Rad23. There is current discussion as to whether XPC or XPA acts first in the repair process to recognise damage or distortions. TFIIH may play a role in distinguishing the damaged strand from the non-damaged one, if translocation along a DNA strand by the TFIIH DNA helicases is interrupted by encountering a lesion. The recognition and incision steps of human NER use 15 to 18 polypeptides, whereas E. coli requires only three proteins to obtain a similar result. Despite this, many remarkable similarities in the NER mechanism have emerged between eukaryotes and bacteria. These include use of a distortion-recognition factor, a strand separating helicase to create an open preincision complex, participation of structure-specific endonucleases and the lack of a need for certain factors when a region containing damage is already sufficiently distorted. PMID- 10675031 TI - Stable and full rescue of the pigmentation in a medaka albino mutant by transfer of a 17 kb genomic clone containing the medaka tyrosinase gene. AB - In the medaka Oryzias latipes, several albino strains have mutations in the tyrosinase gene that have been fully characterized at the molecular level. A genomic clone from wild-type medaka containing the 5 kb tyrosinase gene with its five exons, 10 kb of upstream sequences and 2 kb downstream sequences was introduced into fertilized eggs from a tyrosinase-negative albino strain. We show that the injection of this genomic clone predominantly conferred mosaic expression ending before the hatching stage. A minority of juveniles retained a variable number of pigmented cells, including four individuals keeping one pigmented eye through adulthood. Two of these could be mated, and one of these transmitted the transgene resulting in complete rescue of pigmentation to 16% of its offspring. The resulting transgenic line harbors a single copy of the wild type tyrosinase gene and all fish are wild-type with respect to pigmentation. These experiments suggest that the tyrosinase genomic clone, or a future shorter version of it, can be used in fish to routinely detect transgenic lines. The apparent faithful and systematic expression of the tyrosinase transgene is most probably due to the presence of a locus control region (LCR) in the injected clone. PMID- 10675032 TI - Identification and cloning of an aspartyl proteinase from Coccidioides immitis. AB - A 45 kDa protein was isolated from a soluble vaccine prepared from formaldehyde killed spherules of Coccidioides immitis. From the N-terminal amino acid sequence, the protein yielded a 17-amino-acid peptide that was homologous to sequences of other fungal aspartyl proteinases. The coccidioidal cDNA encoding the proteinase was amplified using oligonucleotide primers designed from the 45 kDa N-terminal amino acid sequence and a fungal aspartyl proteinase consensus amino acid sequence. The PCR product was cloned and sequenced, and the remaining 5' upstream and 3' downstream cDNA was amplified, cloned, and sequenced. The cDNA encoding the coccidioidal aspartyl proteinase open reading frame was cloned and the fusion protein containing a C-terminal His-tag expressed in E. coli. The recombinant aspartyl proteinase was purified by immobilized metal affinity chromatography. This recombinant protein will be used for further studies to evaluate its antigenicity, including protective immunogenicity. PMID- 10675033 TI - Temperature-dependent sex determination in the American alligator: expression of SF1, WT1 and DAX1 during gonadogenesis. AB - Sex determination in mammals and birds is chromosomal, while in many reptiles sex determination is temperature dependent. Morphological development of the gonads in these systems is conserved, suggesting that many of the genes involved in gonad development are also conserved. The genes SF1, WT1 and DAX1 play various roles in the mammalian testis-determining pathway. SF1 and WT1 are thought to interact to cause male-specific gene expression during testis development, while DAX1 is believed to inhibit this male-specific gene expression. We have cloned SF1 and DAX1 from the American alligator, a species with temperature-dependent sex determination (TSD). SF1, DAX1 and WT1 are expressed in the urogenital system/gonad throughout the period of alligator gonadogenesis which is temperature sensitive. SF1 appears to be expressed at a higher level in females than in males. This SF1 expression pattern is concordant with the observed pattern during chicken gonadogenesis, but opposite to that observed during mouse gonadogenesis. Although the observed sexual dimorphism of gonadal SF1 expression in alligators and chickens is opposite that observed in the mouse, it is probable that SF1 is involved in control of gonadal steroidogenesis in all these vertebrates. DAX1 and WT1 are both expressed during stages 22-25 of both males and females. However, there appear to be no sex differences in the expression patterns of these genes. We conclude that DAX1, WT1 and SF1 may be involved in gonadal development of the alligator. These genes may form part of a gonadal development pathway which has been conserved through vertebrate evolution. PMID- 10675034 TI - Organization of the mouse ASGR1 gene encoding the major subunit of the hepatic asialoglycoprotein receptor. AB - The hepatic asialoglycoprotein receptor was the first of the mammalian lectins to be recognized and has been the subject of intense investigation for three decades. Yet, the precise biological role of this major hepatic endocytic receptor has remained elusive. We describe here the characterization of the mouse gene for the major subunit of this receptor (ASGR1) along with 3.5 kb of the upstream 5' region. The gene comprises eight coding exons, with the major transcript in liver displaying a single non-coding 5' exon. A minor hepatic transcript initiates 435 bp upstream of the major start and includes an additional 5' non-coding exon and intron. A minimal 600 bp proximal region of ASGR1 exhibits hepatic-specific promoter activity in HepG2 cells in vitro. These results provide the basis for more detailed genetic studies on the functional role of the hepatic asialoglycoprotein receptor in mammals. PMID- 10675035 TI - Evolutionary conservation of the apolipoprotein E-C1-C2 gene cluster on bovine chromosome 18q24. AB - We have constructed a long-range restriction map spanning about 250 kb on bovine chromosome 18q24. Our results show that the apolipoprotein C2 (APOC2) gene is located about 25 kb from the APOE gene. Four putative CpG islands are also indicated in the map. Interestingly, a minisatellite located in the third intron of the human and mouse APOC2 genes was also found at identical position in the bovine gene and revealed high sequence identity comparing with the two corresponding sequences. By means of cosmid mapping, we further demonstrate that the APOE-APOC1-APOC2 gene cluster is evolutionary conserved in cattle. PMID- 10675036 TI - Two genes encoding a putative multidrug efflux pump of the RND/MFP family are cotranscribed with an rpoH gene in Bradyrhizobium japonicum. AB - The rpoH3 gene of Bradyrhizobium japonicum codes for one of three sigma32-type transcription factors in this organism and is flanked by rag (rpoH3-associated) genes comprising the chromosomal arrangement ragABrpoH3ragCD. The first genes in this cluster code for a classical two-component regulatory system with an unknown function (Narberhaus et al., 1997. Mol. Microbiol. 24, 93-104). The deduced proteins of the last two genes display a high sequence similarity to heavy metal or multidrug efflux pumps of the RND (Resistance/Nodulation/cell Division)-MFP (Membrane Fusion Protein) family. Reverse transcription and polymerase chain reaction (RT-PCR) analysis demonstrated that ragC is cotranscribed with rpoH3. Mutant strains carrying disrupted rag genes or an extented deletion of the rag locus exhibited neither an apparent growth defect nor a deficiency in the symbiotic interaction with soybean roots. The minimal inhibitory concentrations (MICs) of various metals and organic compounds were identical for the wild-type and mutant strains. Moreover, translational lacZ fusions to each of the first four genes of the rag cluster showed a very low expression under all conditions tested; hence, the substrate for the putative efflux pump has not been uncovered. PMID- 10675037 TI - cDNA cloning of run, a Caenorhabditis elegans Runt domain encoding gene. AB - PEBP2/CBF is a heterodimeric transcription factor composed of alpha and beta subunits. The essential roles of mammalian PEBP2alpha genes, PEBP2aA/CBFA1 and PEBP2alphaB/CBFA2 in osteogenesis and hematopoiesis have been well documented. The PEBP2alpha proteins contain a 128 amino acid (aa) region which is highly homologous to Drosophila melanogaster runt and lozenge. The evolutionarily conserved region has been named the Runt domain. In this study, we isolated a cDNA encoding the Caenorhabditis elegans homolog of mammalian PEBP2alpha. The cDNA encodes a 301-aa protein with a highly conserved Runt domain. In addition, a IWRPF five aa motif is present at the C-terminal end. PMID- 10675038 TI - Expression analysis and characterization of the mutant of a growth-phase- and starvation-regulated monofunctional catalase gene from Xanthomonas campestris pv. phaseoli. AB - Analysis of the Xanthomonas campestris pv. phaseoli (Xp) catalase profile using an activity gel revealed at least two distinct monofunctional catalase isozymes denoted Kat1 and Kat2. Kat1 was expressed throughout growth, whereas Kat2 was expressed only during the stationary phase of growth. The nucleotide sequence of a previously isolated monofunctional catalase gene, Xp katE, was determined. The deduced amino acid sequence of Xp KatE showed a high percentage identity to an atypical group of monofunctional catalases that includes the well-characterized E. coli katE. Expression of Xp katE was growth phase-dependent but was not inducible by oxidants. In addition, growth of Xp in a carbon-starvation medium induced expression of the gene. An Xp katE mutant was constructed, and analysis of its catalase enzyme pattern showed that Xp katE coded for the Kat2 isozyme. Xp katE mutant had resistance levels similar to the parental strain against peroxide and superoxide killing at both exponential and stationary phases of growth. Interestingly, the level of total catalase activity in the mutant was similar to that of the parental strain even in stationary phase. These results suggest the existence of a novel compensatory mechanism for the activity of Xp catalase isozymes. PMID- 10675039 TI - Cloning and expression of the mouse Pse gene encoding a novel Ets family member. AB - Human prostate-specific Ets (hPSE) is a novel Ets transcription factor and is exclusively expressed in human prostate glandular epithelium. To explore the role of PSE, we cloned the mouse Pse (mPse) and examined its pattern of expression. A sequence analysis indicated that mPse contains a conserved carboxy-terminal ETS DNA-binding domain and central Pointed domain, and the overall amino acid sequence shares 86% identity with that of hPSE. The ETS DNA-binding domain is highly conserved between human and mouse (98.8% sequence identity) and is similar to Drosophila dets4 (76.7% identity), but not similar to other Ets factors. A Northern blotting analysis revealed that mPse shows organ-specific expression. An in situ hybridization analysis of the prostate and intestine showed that mPse transcripts were present in their epithelial cells. mPse transactivates the promoter of the MASPIN gene in transient transfection assay. These results suggest that mPse encodes a novel Ets family member and is expressed in epithelial cells of restricted organs. PMID- 10675040 TI - Investigation of Schizosaccharomyces pombe as a cloning host for human telomere and alphoid DNA. AB - The fission yeast Schizosaccharomyces pombe (Sch. pombe) has been proposed as a possible cloning host for both mammalian artificial chromosomes (MACs) and mammalian genomic libraries, due to the large size of its chromosomes and its similarity to higher eukaryotic cells. Here, it was investigated for its ability to form telomeres from human telomere sequence and to stably maintain long stretches of alphoid DNA. Using linear constructs terminating in the telomere repeat, T2AG3, human telomere DNA was shown to efficiently seed telomere formation in Sch. pombe. Much of the human telomeric sequence was removed on addition of Sch. pombe telomeric sequence, a process similar to that described in S. cerevisiae. To investigate the stability of alphoid DNA in fission yeast, bacterial artificial chromosomes (BACs) containing 130 and 173 kb of alphoid DNA were retrofitted with the Sch. pombe ars1 element and ura4+ marker using Cre-lox recombination. These alphoid BACs were found to be highly unstable in Sch. pombe deleting down to less than 40 kb, whilst control BACs of 96 and 202 kb, containing non-repetitive DNA, were unrearranged. Alphoid DNA has been shown to be sufficient for human centromere function, and this marked instability excludes Sch. pombe as a useful cloning host for mammalian artificial chromosomes. In addition, regions containing repetitive DNA from mammalian genomes may not be truly represented in libraries constructed in Sch. pombe. PMID- 10675041 TI - Structure and expression pattern of a human MTG8/ETO family gene, MTGR1. AB - AML1-MTG8 fusion protein, which is produced from the rearranged gene formed between AML1 and MTG8 in myeloid leukemia with t(8;21) chromosomal translocation, plays an important role in the pathogenesis of leukemia. We previously showed that ectopically expressed AML1-MTG8 fusion protein is associated with an MTG8 like protein in the mouse myeloid precursor cell line L-G, and this association seemed to be required for AML1-MTG8 to stimulate proliferation. As a candidate cDNA for this MTG8-like protein, a 6.4 kb MTGR1 cDNA encoding human MTGR1b protein of 604 amino acids was isolated. Since this cDNA was shorter than the main mRNA (about 7.5 kb), the 5'-end of the MTGR1 cDNA was extended using Marathon Ready cDNA. When the newly obtained 5'-sequence was combined with the previous cDNA, the resultant MTGR1 cDNA (6995 bp), including exon 3 that the previous cDNA lacked, could encode MTGR1a protein of 575 amino acids. Transcripts of the MTGR1 gene were expressed ubiquitously in the human tissues and cell lines examined. PCR analyses of the cDNAs from human tissues showed the presence of various splicing variants with regard to the 5'-region including exons 1, 2 and 3. The MTGR1 gene consists of 14 exons and spans about 68 kb. The genomic structure of MTGR1 is highly similar to those of other MTG 8-family genes, MTG8 and MTG16. MTG16 was recently cloned from the translocation breakpoint of myeloid malignancies with t(16;21) chromosomal translocation. PMID- 10675042 TI - Identity between rat htf and human xbp-1 genes: determination of gene structure, target sequence, and transcription promotion function for HTF. AB - Hepatocarcinogenesis-related transcription factor (HTF) was originally isolated from rats in which the expression was enhanced in hepatocellular carcinomas. Rat HTF (rHTF) is structurally similar to human X-box-binding protein-1 (hXBP-1), and both factors are unique in respective genomes. A previous study showed that hXBP 1 mRNA is detectable ubiquitously but is enriched in the human liver as rHTF. In this study, we demonstrated the analogous exon-intron organization and significant sequence homology for rhtf and hxbp-1 genes. Alignment of amino acid sequences of rHTF and hXBP-1 revealed that all the characteristic motifs in rHTF were conserved in hXBP-1. Moreover, Southern blotting patterns provided with the rHTF and hXBP-1 probes were basically the same. These two genes were thus thought to belong to the same evolutional lineage. We determined the consensus binding sequence (CRCGTCA) for rHTF by CASTing, and it was found to be nearly the same as that for hXBP-1. Transactivation ability of rHTF was also demonstrated. The rhtf gene generates two types of mRNAs (2.0 kb and 2.5 kb), both of which encode identical rHTF protein. These transcripts had distinct transcription initiation sites. The 2.0 kb promoter, that was revealed by the transient luciferase assay, contained GC-box and CAAT-box. Sequences around the transcription initiation site for the 2.0 kb transcript were similar in rhtf and hxbp-1 genes. Our observations suggest that HTF is a rat homolog of hXBP-1. PMID- 10675043 TI - A highly representative two-hybrid genomic library for the yeast Yarrowia lipolytica. AB - Since its description by Fields and Song in 1989 (Nature 340, 245-246), the yeast two-hybrid system has been used extensively to study protein-protein interactions, becoming increasingly efficient with technological and methodological improvements. Here, we report the construction of a highly representative two-hybrid genomic library for the dimorphic yeast Yarrowia lipolytica based on the system described by James et al. (1996. Genetics 144, 1425-1436). The endoplasmic reticulum protein Slslp was then used as a bait in a functional test of the library. Indeed, we previously showed that the SLS1 gene product is involved in protein translocation across the endoplasmic reticulum membrane and interacts physically in a two-hybrid assay with Kar2p, an essential luminal member of the HSP70 family (Boisrame et al., 1998. J. Biol. Chem. 273, 30 903-30 908). We developed a mating strategy similar to that used for the Saccharomyces cerevisiae FRYL library (Fromont-Racine et al., 1997. Nat. Genet. 16, 277-282). No other partner than Kar2p was identified in this screen. As an interesting result, Kar2p interacts with Slslp through its ATPase domain, supporting our hypothesis that Slslp is a cofactor of the chaperone protein, modulating its activity during the HSP70 cycle. Our results indicate that we have constructed a new and powerful tool for the study of Yarrowia lipolytica, which we believe is a good alternative model to investigate such complex biological processes as secretion pathways. PMID- 10675044 TI - A gene whose major transcript encodes only the substrate-binding domain of a protein-tyrosine kinase. AB - We have identified a novel protein-tyrosine kinase gene family in the simple multicellular animal Hydra vulgaris that consists of at least three members. Two of the genes encode receptor protein-tyrosine kinases. The third member of the family is unusual in that in non-sexual animals, the only transcripts that it produces encode polypeptides lacking all or nearly all of the ATP-binding lobe. Characterization of multiple cDNA clones and hybridization mapping of genomic DNA indicate that the gene, which we have termed Hinterteil (Hint), undergoes alternative cis-splicing, alternative trans-splicing, and alternative polyadenylation. In-situ hybridization analysis shows that expression of the gene is upregulated during spermatogenesis. Sexual males also produce an additional Hint transcript that is larger than the transcript seen in non-sexual animals, but still not large enough to encode a receptor. PMID- 10675045 TI - Selection and genetic drift of polymorphisms within the merozoite surface protein 1 gene of Plasmodium falciparum. AB - Intragenic recombination in the merozoite surface protein-1 gene (Msp-1) of Plasmodium falciparum is a major mechanism for allelic variation among natural parasite populations. The frequency of recombination depends on the intensity of transmission in the vector mosquito. In the present study, linkage disequilibrium between polymorphic 'loci' in the 5'- and 3'-regions of Msp-1 was examined in parasite populations from Brazilian Amazon and southern Vietnam and compared with that in a Thai population previously reported. The R2 test identified clusters of linkage disequilibria between the 5'- and 3'-regions, which are different among the three populations. However, the overall strength of linkage disequilibria was stronger in Brazil, a hypoendemic area, than in Vietnam and Thailand, mesoendemic areas, suggesting that linkage disequilibrium in Msp-1 inversely correlates with the intensity of transmission. To investigate possible mechanisms for linkage disequilibrium in Msp-1, we applied the Fst index, which measures the inter population variance in allele frequency, to 'loci' in Msp-1 among the three populations. The Fst test identified two distinct regions with respect to inter population allele frequency in Msp-1: one for highly divergent 'loci' in the 5' region and the other for non-divergent 'loci' in the 3'-region. These results suggest that genetic drift is not the sole mechanism for linkage disequilibrium, but selection operates on 'loci' in the 3'-region in hypo- and mesoendemic areas of malaria. PMID- 10675046 TI - S phase-specific DNA-binding proteins interacting with the Hex and Oct motifs in type I element of the wheat histone H3 promoter. AB - The type I element (CCACGTCANCGATCCGCG), consisting of the Hex motif (CCACGTCA) and the reverse-oriented Oct motif (GATCCGCG), is necessary and sufficient to confer the S phase-specific transcription of the wheat histone H3 (TH012) gene. The transcriptional regulation via the type I element is thought to occur through interactions between transcription factors which bind specifically to the Hex and Oct motifs. Here we report S phase-specific DNA-binding proteins interacting with the type I element in partially synchronized wheat cultured cells. Hex motif binding proteins found here resembled HBP-1a, as reported previously in terms of DNA-binding specificity. DNA-binding activities of the HBP-1a-like proteins were modulated by phosphorylation/dephosphorylation. In the electrophoretic mobility shift assay of the wheat nuclear extract, we also found three Oct motif-specific binding proteins, named OBRF (octamer-binding regulatory factor)-1, -2 and -3. One of the HBP-1a-like proteins and OBRF-1 appeared predominantly at the S phase. Thus, it was supposed that these two factors play a crucial role in the S phase specific regulation of wheat histone gene expression. PMID- 10675047 TI - Finding ways to create connections among communities: partial results of an ethnography of urban public health nurses. AB - The purpose of this ethnographic study was to describe the culture of public health nurses (PHNs) in a large, Midwestern urban health department. Data collection methods, data management, and analyses followed ethnographic procedures and resulted in the development of categories, domains, and cultural themes. The general study participants were PHNs, clients, supervisors, and administrators. The primary cultural theme that emerged was that public health nursing is finding ways to create connections among communities. Three interacting communities were identified: the local communities, communities created by individuals and families, and communities of resources. This article describes one of the three subthemes that emerged, processes used to help clients create connections, and describes how caring is shown uniquely in public health nursing. As a result of the study, implications for nursing practice, education, and research were developed. The results of the study supported a position that public health nursing is a unique nursing specialty. It reinforced also the applicability of an ethnographic design and methodology to nursing research. PMID- 10675048 TI - Cudahy high school survey and focus groups: assessment of the needs of a teen population. A community-campus collaboration. AB - Collaboration between local public health agencies and university schools of nursing can be advantageous to both parties. Students need opportunities to learn aggregate-based care; health officers need community partnerships that expand their potential to accomplish core functions. This article offers a case study to illustrate a collaborative relationship. A high school survey and a plan for teen services were the focus of the joint endeavor. With guidance from faculty, students offered labor and expertise; the agency offered a real world laboratory for learning. PMID- 10675049 TI - The relationship between self-esteem, health habits, and knowledge of BSE practice in female inmates. AB - This article reports on data derived from an investigation of the self-esteem, health habits, and knowledge of breast self-examination (BSE) practice in female inmates. A descriptive correlational study was conducted with a prison sample of 197 adult females incarcerated in a women's state prison. Major findings of the study suggest that female inmates in general had medium to high self-esteem, poor health habits before incarceration, and minimal knowledge about BSE practice. Only 26% reported correct knowledge related to frequency in BSE practice, and few reported that they knew correct BSE technique. In addition, findings suggest that a correlation does not exist between self-esteem and knowledge of BSE practice, and participants' last grade completed served as a good predictor of women's knowledge of BSE practice. PMID- 10675050 TI - Gender issues and Japanese family-centered caregiving for frail elderly parents or parents-in-law in modern Japan: from the sociocultural and historical perspectives. AB - This paper presents a sociocultural and historical literature review of gender related issues associated with family-centered caregiving for frail, elderly relatives in modern Japan. Issues addressed from a Japanese perspective are (a) women and social norms of caregiving, (b) feminine identity and caregiving, (c) women in the workforce, and (d) women and caregiving. Implications for research are also discussed. PMID- 10675051 TI - Validating the safety of nurse-health advocate services. AB - Prior to promoting the use of community health care advocates for home visits, it is necessary to evaluate their ability to safely screen for health problems. This study examined trained maternal-child health advocates (MCHAs) who, supervised by professional nurses, conducted maternal-child home visits consisting of health promotion and problem identification. Problems identified by MCHAs were compared to problems identified by professional, validating nurses, who were not part of the service project, on hypothetical home visit situations and during 213 duplicate home visits. There were no significant differences between MCHAs and professional nurses in their identifications of infant health problems, infant health care deficits, other family members' health problems, prenatal care deficits, emotional problems, and substance abuse on either the hypothetical home visits or the duplicate home visits. The validating nurses identified significantly more women's health problems (p = 0.01) and women's health care deficits (p = 0.02) than the MCHAs on the duplicate home visits. These findings validate the safety of using the model of trained community health advocates teamed with registered nurses to screen for infant health problems during home visits. PMID- 10675052 TI - Personal safety, violence, and home health. AB - A critical issue facing the health care industry today is the potential impact of community and interpersonal violence on home health care. The purposes of this study were to (1) serve as a source for understanding the personal safety risk issues facing home care staff in a large Midwest region and its surrounding rural areas; (2) provide an understanding of how perceived threats to personal safety may impact patient care and patient outcomes; (3) identify strategies for increasing the personal safety of direct care staff; and (4) identify organizational, educational, and procedural issues that impede or enhance staff safety. A triangulated qualitative design was used including focus groups, in depth individual interviews, critical event narratives, and a participant self report form. The study used a purposive sample consisting of 5 men and 56 women who were either administrators or direct care staff from 13 home health agencies. Seven major themes emerged: (1) unsafe conditions that direct care staff must face; (2) organizational and administrative issues that impede or promote the personal safety of staff; (3) ethical issues staff face daily; (4) protective factors associated with maintaining safety; (5) issues of gender, race, age, and experience; (6) education and training; and (7) the potential impact that staff's fear of interpersonal and community violence can have on patient care and patient outcomes. PMID- 10675053 TI - Prevention strategies other than male condoms employed by low-income women to prevent HIV infection. AB - This study sought to determine HIV prevention strategies other than male condom use employed by low-income women who have sex with men (WSM) and to identify variables that predict use of these strategies. A cross-sectional survey of nearly 4,000 women receiving Women, Infants, and Children (WIC) benefits in 21 Missouri counties was conducted. The response rate was 58%, with 2,256 completed questionnaires returned. Women were asked to indicate one or more of nine methods they had ever used to prevent HIV infection. Women were also asked about their use of male condoms, preference for male condoms versus female condoms, and which partner usually made decisions about STD/HIV prevention. Of the 2,256 questionnaires returned, 1,325 WSM indicated use of at least one HIV prevention strategy other than condom use. Strategies were: being tested for HIV (68.2%), partner being tested for HIV (44.1%), asking partner about his sex history (41.1%), using oral contraceptives (18.8%), asking him if he has HIV (13.7%), douching (11.8%), withdrawal (9.4%), and having anal or oral sex (6.6%). Common predictors of these strategies were race, education, history of STD, condom use, and marital status. Basic misunderstandings about HIV prevention are common in specified subpopulations of low-income women. HIV prevention programs for low income WSM should capitalize on women's efforts to prevent HIV by designing programs to help women replace ineffective prevention strategies with effective prevention strategies. PMID- 10675054 TI - A case study in translation methodology using the Health-Promotion Lifestyle Profile II. AB - Although Hispanics constitute the most rapidly growing segment of the population in the United States, they have received relatively little attention regarding factors affecting their health behaviors and influences. One such factor is the scarcity of reliable and valid Spanish-language instruments for research with this population. Researchers who attempt to translate an existing instrument into Spanish need to recognize the methodological issues involved in the translation process and psychometric testing. The purpose of this article is to describe the advantages and disadvantages of various translation methodologies, to identify statistical issues in cross-cultural research, and to provide a case study of the translation process and statistical analysis of a translated instrument. Specifically, this study looks at the development and pilot testing of a Spanish language version of the Health-Promoting Lifestyle Profile II using a randomized convenience sample of 60 bilingual Hispanic individuals. PMID- 10675055 TI - An investigation of perceived exertion via whole body exertion and direct muscle force indicators during the determination of the maximum acceptable weight of lift. AB - The objective of this study was to identify the perceived exertion mechanisms (direct muscle force and whole body exertion) associated with the decision to change the weight of lift during the determination of the maximum acceptable weight of lift (MAWL). Fifteen males lifted a box of unknown weight at a rate of 4.3 lifts/min, and adjusted the weight until their MAWL was reached. Variables such as the predicted muscle forces and heart rate were measured during the lifting exertion, as well as the predicted spinal loading in three dimensions using an EMG-assisted biomechanical model. Multiple logistic regression techniques were used to identify variables that were associated with the decision to change the weights up and down prior to a subsequent lift. Results indicated that the force in the left erector spinae, right internal oblique, and left latissimus dorsi muscles as well as heart rate were associated with decreases in the weight prior to the next lift. It appears that a combination of local factors (muscle force) and whole body exertion factors (heart rate) provide the feedback for the perceived exertion when decreasing the weight. The up-change model indicated that the forces of the right erector spinae, left internal oblique, and the right latissimus dorsi muscles were associated with the decision to increase the weight prior to the next lift. Thus, local factors provide feedback during the decision to increase the weight when starting from light weights. Collectively, these findings indicate that psychophysically determined weight limits may be more sensitive to muscular strain rather than spinal loading. PMID- 10675056 TI - Calculation of times to exhaustion at 100 and 120% maximal aerobic speed. AB - The aim was to compare physiologic responses during exhaustive runs performed on a treadmill at 100 and 120% maximal aerobic speed (MAS: the minimum speed that elicits VO2max). Fourteen subelite male runners (mean +/- SD; age = 27+/-5 years; VO2max = 68.9+/-4.6 ml/kg(-1)/min(-1); MAS = 21.5+/-1 km/h(-1)) participated. Mean time to exhaustion tlim100% at 100% MAS (269+/- 77s) was similar to those reported in other studies. However, there was large variability in individual tlim100% MAS (CV = 29%). MAS was positively correlated with VO2max (r = 0.66, p<0.05) but not with tlim100%) MAS (r = -0.50, p<0.05). tlim100% MAS was correlated with t(lim) at 120% MAS (r = 0.52, p < 0.05) and to blood pH following the rest at 120% MAS (r = -0.68, p<0.05). The data suggest that running time to exhaustion at MAS in subelite male runners is related to time limit at 120% (tlim120%) MAS. Moreover, anaerobic capacity determined by the exercise to exhaustion at 120% MAS can be defined as the variable 'a' in the model of Monod and Scherrer (1954). PMID- 10675057 TI - Optimum seat pan and back-rest parameters for a comfortable tractor seat. AB - An experimental set up was fabricated to measure the pressure distribution on the seat pan and back-rest of a tractor seat. Experiments were conducted with four different seat pans having radius of curvatures of 60, 75, 90 and infinity cm, four back-rests with radius of curvatures of 30, 60, 90 and infinitity cm, and three back-rest inclinations of 0 degrees , 5 degrees and 10 degrees on representative Indian tractor operators. The subjective assessment of perceived comfort at the seat-operator interface was also recorded. Experiments were conducted in a randomized block design and the data obtained were analysed using suitable computer packages. Results indicate that all the parameters, namely seat pan, back-rest profile curvatures and the back-rest angle of inclination, affect the pressure distribution. It is concluded that a seat pan with radius of curvature 75 cm, back-rest with radius of curvature 30 cm and back-rest inclination of 10 degrees are the most suitable parameters for Indian tractor operators. PMID- 10675058 TI - On-line driver workload estimation. Effects of road situation and age on secondary task measures. AB - In order to develop a driver-car interface that adapts the presentation of messages generated by in-vehicle information systems to driver workload, two experiments investigated potential determinants of driver visual and mental workload as indicated by performance on two secondary tasks. Experiment 1 suggested that road situation is a major determinant of visual and mental workload of the driver and that the processing resources of older drivers are somewhat more limited than those of younger and middle-aged drivers. Familiarity with the area of driving (when guided) and time of day (associated with traffic density) showed no secondary task effects. Experiment 2 showed that the categorization of road situations, proposed in Experiment 1, could underlie adaptation of visually loading messages to the workload incurred by driving. This was not found with respect to mentally loading messages. PMID- 10675059 TI - The effects of hyperoxia on performance during simulated firefighting work. AB - This study evaluated the effects of hyperoxia (inspired oxygen fraction = 40%) on performance during a simulated firefighting work circuit (SFWC) consisting of five events. On separate days, 17 subjects completed at least three orientation trials followed by two experimental trials while breathing either normoxic (NOX) and hyperoxic (HOX) gas mixtures that were randomly assigned in double-blind, cross-over design. Previously, ventilatory threshold (Tvent) and VO2max had been determined during graded exercise (GXT) on a cycle ergometer. Lactate concentration in venous blood was assessed at exactly 5 min after both the experimental trials and after the GXT. Total time to complete the SFWC was decreased by 4% (p < 0.05) with HOX. No differences were observed in individual event times early in the circuit, however HOX resulted in a 12% improvement (p < 0.05) on the final event. A significantly decreased rating of perceived exertion (RPE) was also recorded immediately prior to the final event. No differences were observed in mean heart rate or post-exercise blood lactate when comparing NOX to HOX. Heart rates during the SFWC (both conditions) were higher than HR at Tvent, but lower than HR at VO2max (p<0.05). Post-SFWC lactate values were higher (p<0.05) than post-VO2max. These results demonstrate that hyperoxia provided a small but significant increase in performance during short duration, high intensity simulated firefighting work. PMID- 10675060 TI - Road-edge delineation in rural areas: effects on driving behaviour. AB - When driving on lower-category Dutch rural roads without any delineation, drivers are likely to drift off the road with their right-side wheels, thus incurring damage to the pavement edge or even leading to accidents. In two experiments, two types of road-edge delineation, with continuous or dashed edge lines, were compared with two control roads without lines or with only a dashed line on the road axis. The first experiment consisted of non-obtrusive video recordings of passing traffic. Vehicle position on the experimental roads was more to the road's centre than on the control roads. The second experiment was a driving test with an instrumented vehicle, during daytime lighting and during darkness. Again, vehicle lateral position was more central on the experimental roads, especially during darkness. Subjects could safely pass oncoming vehicles. Driving speed increased on the experimental roads compared with the unlined control road, but not beyond speeds found on the axis-lined control road. Driver's mental effort while driving over the experimental roads did not differ from the effort while driving over the control roads. Subjectively rated effort was higher for the unlined control road than for the three other roads. Subjects preferred the edge lined roads to the unlined control road, but not more than the axis-lined control road. It was concluded that edge-lines may provide a simple and effective way of inducing a more favourable lateral position on rural roads without having negative effects on subjective appraisal, driving performance or mental workload. PMID- 10675061 TI - Temporal variation in the luminance level of stimuli displayed on a cathode-ray tube monitor: effects on performance on a visual vigilance task. AB - This paper assesses the extent to which the sensitivity decrement frequently observed in vigilance tasks is affected by temporal variations in the luminance level of the stimuli displayed on the screen of a cathode-ray tube (CRT) monitor. First, it was confirmed that the luminance of the stimuli displayed on the screen of the CRT monitor decreases substantially during the first hour after turning the monitor on, and then it remains quite stable. Second, an experiment was carried out in which participants performed a visual vigilance task at three different time periods within which the luminance of the stimuli displayed on the screen of a CRT monitor either decreased or remained stable. The results indicate that the vigilance decrement is modulated by temporal fluctuations of the luminance of the monitor screen, which is used to display the stimuli. However, the relationship between both variables is not simple: the largest sensitivity decrement was not associated with the largest luminance decrement, but to a medium luminance decrement. PMID- 10675062 TI - Perceived fatigue after mental work: an experimental evaluation of a fatigue inventory. AB - The aim of this study was to investigate the perceived fatigue after mental work and to test the Swedish Occupational Fatigue Inventory (SOFI). Twenty male and 20 female participants worked with proof reading (2 x 90 min) and a vigilance task (2 x 60 min). After each task session, perceived fatigue was rated with the SOFI and Borg's CR10O-scale. In addition, physiological reactions were registered; blood pressure, heart rate, heart rate variability and muscle activity in corrugator supercilii, as well as measures of performance: reaction time, number of pages read and number of proof errors found, number of detected signals. As expected, the highest ratings were obtained on Lack of energy, Lack of motivation and Sleepiness, particularly after the vigilance task. High ratings after both work tasks were also found on the CR10-scale. Men and women did not differ significantly with respect to their ratings. No clear-cut physiological reactions were found to correlate with ratings of fatigue. The results indicate the validity of the mental dimension of the SOFI. PMID- 10675063 TI - Gender differences in exerted forces and physiological load during pushing and pulling of wheeled cages by postal workers. AB - The aim was to determine gender differences regarding exerted forces and physiological load during push/pull tasks simulating the daily working practice of postal workers. Eight female and four male workers handled four-wheeled cages under eight conditions corresponding to the cage weight (130, 250, 400, 550 kg) and the direction of force exertion (pushing, pulling). For each of the five dependent variables, average force, initial force, ending force, oxygen uptake and heart rate, two analyses of variance with repeated measurements were performed, i.e. with and without correction for the worker's body weight, body height and maximum capacity regarding the dependent variable. Exerted forces and physiological load were high for the cages weighing 400 and 550 kg. Gender differences were significant for all dependent variables (p = 0.030-0.000). When the personal factors were included in the model, male workers exerted significantly higher average forces and ending forces than their females, while differences regarding initial forces and physiological load were not significant. However, none of the personal factors were significantly related to any of the dependent variables. It is concluded that gender differences in exerted forces were not caused by differences in anthropometry and maximum capacity, but due to application of different work methods by women in order to balance work demands and work ability. PMID- 10675064 TI - A survey of hand anthropometry of female rural farm workers in Ibadan, western Nigeria. AB - An anthropometric survey measuring 18 dimensions of the right hand in 37 female rural farm workers living in Ibadan, western Nigeria was conducted. The means, standard deviations and percentile values are reported for these. The means of the collected data are compared with those for females from the UK, from Hong Kong and from America, using data from other published studies. The results suggest that the Nigerian female hand is wider and thicker, but shorter than that of their foreign counterparts. Such differences have implications for design and evaluation of hand tools for the Nigerian female population. PMID- 10675065 TI - When doctors become agents of the state. PMID- 10675066 TI - HOPE and extension of the indications for ACE inhibitors? Heart Outcomes Prevention Evaluation. PMID- 10675067 TI - Fluoride and fractures: an ecological fallacy. PMID- 10675068 TI - New insights into role of microenvironment in multiple myeloma. PMID- 10675069 TI - Perineal massage for prevention of perineal trauma in childbirth. PMID- 10675070 TI - Antenatal corticosteroids: is more better? . PMID- 10675071 TI - Effects of ramipril on cardiovascular and microvascular outcomes in people with diabetes mellitus: results of the HOPE study and MICRO-HOPE substudy. Heart Outcomes Prevention Evaluation Study Investigators. AB - BACKGROUND: Diabetes mellitus is a strong risk factor for cardiovascular and renal disease. We investigated whether the angiotensin-converting-enzyme (ACE) inhibitor ramipril can lower these risks in patients with diabetes. METHODS: 3577 people with diabetes included in the Heart Outcomes Prevention Evaluation study, aged 55 years or older, who had a previous cardiovascular event or at least one other cardiovascular risk factor, no clinical proteinuria, heart failure, or low ejection fraction, and who were not taking ACE inhibitors, were randomly assigned ramipril (10 mg/day) or placebo, and vitamin E or placebo, according to a two-by two factorial design. The combined primary outcome was myocardial infarction, stroke, or cardiovascular death. Overt nephropathy was a main outcome in a substudy. FINDINGS: The study was stopped 6 months early (after 4.5 years) by the independent data safety and monitoring board because of a consistent benefit of ramipril compared with placebo. Ramipril lowered the risk of the combined primary outcome by 25% (95% CI 12-36, p=0.0004), myocardial infarction by 22% (6-36), stroke by 33% (10-50), cardiovascular death by 37% (21-51), total mortality by 24% (8-37), revascularisation by 17% (2-30), and overt nephropathy by 24% (3-40, p=0.027). After adjustment for the changes in systolic (2.4 mm Hg) and diastolic (1.0 mm Hg) blood pressures, ramipril still lowered the risk of the combined primary outcome by 25% (12-36, p=0.0004). INTERPRETATION: Ramipril was beneficial for cardiovascular events and overt nephropathy in people with diabetes. The cardiovascular benefit was greater than that attributable to the decrease in blood pressure. This treatment represents a vasculoprotective and renoprotective effect for people with diabetes. PMID- 10675072 TI - Long-term results of overlapping anterior anal-sphincter repair for obstetric trauma. AB - BACKGROUND: Anterior structural damage to the anal sphincter occurs in up to a third of women at first vaginal delivery, and of these a third have new bowel symptoms. The standard treatment for such structural damage is anterior overlapping anal-sphincter repair. We aimed to assess the long-term results of this operation. METHODS: We assessed the long-term results in 55 consecutive patients who had had repair a minimum of 5 years (median 77 months [range 60-96]) previously. Questionnaire and telephone interview assessed current bowel function and continence, restriction in activities related to bowel control, and overall satisfaction with the results of surgery. 42 of these patients had been continent of solid and liquid stool at a median of 15 months after the repair. FINDINGS: We were able to contact 47 (86%) of the 55 patients. One of these patients had required a proctectomy and end ileostomy for Crohn's disease. Of the remaining 46 patients, 27 reported improved bowel control without the need for further surgery, and 23 rated their symptom improvement as 50% or greater. Seven patients had undergone further surgery for incontinence and one patient had not had a covering stoma closed. Thus, the long-term functional outcome of the sphincter repair alone could be assessed in 38 patients. Of these patients, none was fully continent to both stool and flatus; only four were totally continent to solid and liquid stool; six had no faecal urgency; and eight had no passive soiling. Of the 38 patients, 20 still wore a pad for incontinence and 25 reported lifestyle restriction. 14 reported the onset of a new evacuation disorder after sphincter repair. 23 of the 46 patients contacted had a successful long-term outcome (defined as no further surgery and urge faecal incontinence monthly or less). INTERPRETATION: The results of overlapping sphincter repair for obstetric anal sphincter damage seem to deteriorate with time. Preoperative counselling should emphasise that although most patients will improve after the procedure, continence is rarely perfect, many have residual symptoms, and some may develop new evacuation disorders. PMID- 10675073 TI - Fluoride in drinking water and risk of hip fracture in the UK: a case-control study. AB - BACKGROUND: Although the benefits of water fluoridation for dental health are widely accepted, concerns remain about possible adverse effects, particularly effects on bone. Several investigators have suggested increased rates of hip fracture in places with high concentrations of fluoride in drinking water, but this finding has not been consistent, possibly because of unrecognised confounding effects. METHODS: We did a case-control study of men and women aged 50 years and older from the English county of Cleveland, and compared patients with hip fracture with community controls. Current addresses were ascertained for all participants; for those who agreed to an interview and who passed a mental test, more detailed information was obtained about lifetime residential history and exposure to other known and suspected risk factors for hip fracture. Exposures to fluoride in water were estimated from the residential histories and from information provided by water suppliers. Analysis was by logistic regression. FINDINGS: 914 cases and 1196 controls were identified, of whom 514 and 527, respectively, were interviewed. Among those interviewed, hip fracture was strongly associated with low body-mass index (p for trend <0.001) and physical inactivity (p for trend <0.001). Estimated average lifetime exposure to fluoride in drinking water ranged from 0.15 to 1.79 ppm. Current residence in Hartlepool was a good indicator for high lifetime exposure to fluoride. After adjustment for potential confounders, the odds ratio associated with an average lifetime exposure to fluoride > or =0.9 ppm was 1.0 [95% CI 0.7-1.5]. INTERPRETATION: There is a low risk of hip fracture for people ingesting fluoride in drinking water at concentrations of about 1 ppm. This low risk should not be a reason for withholding fluoridation of water supplies. PMID- 10675074 TI - Hormone replacement therapy and accuracy of mammographic screening. AB - BACKGROUND: Hormone replacement therapy (HRT) is commonly used and may affect the accuracy of mammographic screening. METHODS: We examined the sensitivity, specificity, and small-cancer detection rate according to HRT use in 103,770 women in Victoria, Australia, who attended first-round screening in 1994 and who did not have a personal history of breast cancer or a breast lump or a bloodstained or watery nipple discharge at the time of screening. BreastScreen Victoria provides mammography to women aged 40 years and older every 2 years. Unconditional logistic modelling was used to adjust for age, family history, and symptom status. FINDINGS: The sensitivity of screening mammography for a 2-year screening interval was lower in HRT users (64.8% [95% CI 58-72]) than non-users (77.3% [74-81]). In the target group (50-69 years), the sensitivity was 64.3% (57 72) in HRT users and 79.8% (76-84) in non-users. Among women who were diagnosed with cancer during the 2-year screening interval, HRT users were more likely to have a false negative result than non-users (odds ratio 1.60 [1.04-2.21]) after adjusting for potential confounding factors. Specificity was 0.6% lower in HRT users compared with non-users. Among women who did not have cancer diagnosed in the interval, HRT users were more likely to have a false positive result (adjusted odds ratio 1.12 [1.05-1.19]). INTERPRETATION: We show that HRT use reduces the sensitivity of mammographic screening. In countries where HRT use is widespread, the reduction in sensitivity with HRT use may undermine the capacity of population-based mammographic-screening programmes to realise their potential mortality benefit. PMID- 10675075 TI - Infectiousness of Mycobacterium tuberculosis in HIV-1-infected patients with tuberculosis: a prospective study. AB - BACKGROUND: Previous studies concerning the relative infectiousness of HIV-1 positive individuals with pulmonary tuberculosis have produced conflicting results. Thus, we assessed the effect of HIV-1 on the infectiousness of Mycobacterium tuberculosis in a prospective study. METHODS: We organised in Santo Domingo, Dominican Republic, a cohort study of household contacts of HIV-1 positive and HIV-1-negative individuals with newly diagnosed pulmonary tuberculosis. Household contacts were assessed at their houses at baseline and followed up for 14 months for evidence of M tuberculosis infection and tuberculosis with a multi-step tuberculin skin test, anergy skin test, physical examinations, chest radiographs, and sputum smears. FINDINGS: Tuberculin induration of 5 mm or greater was seen in 153 (61%) of 252 household contacts of HIV-1-positive index cases and in 418 (76%) of 551 household contacts of HIV-1 negative index cases (odds ratio 0.49 [95% CI 0.35-0.67], p=0.00001). In multivariate logistic-regression analysis after allowance for between-household variation in tuberculin response, HIV-1 infection of the index case remained inversely associated with the tuberculin response of the household contacts (0.52 [0.29-0.93], p=0.02). When the analysis was restricted to household contacts aged between 2 years and 15 years the adjusted association remained significant (0.37 [0.14-0.98], p=0.04). Among household contacts who had a negative tuberculin skin test at baseline, conversion to tuberculin skin test positivity was less frequent among household contacts of HIV-1-positive index cases (cut-off > or =5 mm: 32/131 [24%] vs 71/204 [35%], p=0.05; cut-off > or =10 mm: 23/153 [15%] vs 55/245 [22%], p=0.07). INTERPRETATION: These data suggest that HIV-1-positive individuals with tuberculosis are less likely than HIV-1-negative individuals with tuberculosis to transmit M tuberculosis to their close contacts. No changes in the current policy regarding tuberculosis contact tracing are needed in the presence of HIV-1. PMID- 10675076 TI - Colitis associated with docetaxel-based chemotherapy in patients with metastatic breast cancer. AB - BACKGROUND: Docetaxel and vinorelbine as combined treatment for metastatic breast cancer can have the dose-limiting toxic effects of mucositis and neutropenic fever. We report unexpected ischaemic colitis in six patients associated with docetaxel-based therapy, three of whom were treated in a phase I study designed to establish the maximum tolerated dose of this combination with the prophylactic use of granulocyte-colony-stimulating factor. METHODS: Between August, 1997, and December, 1998, 14 patients with metastatic breast cancer were treated with vinorelbine, docetaxel, and granulocyte-colony-stimulating factor in a phase I study. Three patients developed colitis similar to that seen in typhlitis. Three additional patients were identified during scheduled review of toxic effects in patients participating in clinical trials involving docetaxel. FINDINGS: Three patients on combined vinorelbine and docetaxel developed colitis-like symptoms. Two patients died, one from necrotic bowel and the other from neutropenic fever and colitis. Two of the patients presented on day 7 and day 8 of chemotherapy, respectively, with neutropenic fever and abdominal pain; the third patient developed neutropenia without fever and abdominal pain on day 8. The other three patients were treated with docetaxel, docetaxel and pamidronate disodium, or docetaxel and cyclophosphamide. All three patients presented with abdominal pain on days 10, 5, and 4, respectively. One had non-neutropenic fever, another had neutropenic fever, and the third was afebrile and non-neutropenic at the time of presentation with abdominal pain. Three patients had blood in their diarrhoea, abdominal tenderness, or both. Computed tomography of the abdomen and pelvis showed features of colitis in three patients. INTERPRETATION: This serious complication may result from the use of docetaxel and may be exacerbated by its combination with vinorelbine. Study of hospital-based patients treated with taxane-based chemotherapy is underway to find out the frequency of such complications. PMID- 10675077 TI - A cough, then respiratory failure. PMID- 10675078 TI - Impact of immunisation on pertussis transmission in England and Wales. AB - Pertussis immunisation reduces disease frequency, but is not thought to prevent transmission. We show that vaccination has substantially reduced transmission in England and Wales. PMID- 10675079 TI - Autoimmune T cells as potential neuroprotective therapy for spinal cord injury. AB - Autoimmune T cells against central nervous system myelin associated peptide reduce the spread of damage and promote recovery in injured rat spinal cord, findings that might lead to neuroprotective cell therapy without risk of autoimmune disease. PMID- 10675080 TI - Structured treatment interruptions to control HIV-1 infection. AB - Structured treatment interruptions progressively lowered the rate of viral rebound in some HIV-1 infected patients. This approach should be explored as an alternative to continuous antiretroviral therapies. PMID- 10675081 TI - Paclitaxel hypersensitivity reactions related to bee-sting allergy. AB - Patients with a history of beesting allergy may have a higher risk of a hypersensitivity reaction with paclitaxel treatment. We suggest careful screening of patients for allergies. PMID- 10675082 TI - Neuroleptic malignant syndrome after venlafaxine. AB - A patient developed neuroleptic malignant syndrome after a single dose of venlafaxine with trifluoperazine treatment. A dopamine-inhibition effect induced by one dose of venlafaxine may have augmented dopamine-receptor inhibition by trifluoperazine. PMID- 10675083 TI - Corneal opacities after cataract surgery with hypromellose. AB - 26 cases of corneal opacity after cataract surgery occurred in a period of 2 weeks in one surgical unit. Cases occurred after a change in source of intraocular hypromellose solution and only among patients in whom the new product had been used. PMID- 10675084 TI - Russian biomedical science survives against all odds. PMID- 10675085 TI - Building capacity for primary care in Tajikistan. PMID- 10675086 TI - Mitochondrial respiratory chain disorders I: mitochondrial DNA defects. AB - Mitochondria have a pivotal role in cell metabolism, being the major site of ATP production via oxidative phosphorylation (OXPHOS); they have a critical role in apoptotic cell death; and they also contribute to human genetics since mitochondria have a functional genome separate from that of nuclear DNA. Defects of mitochondrial metabolism are associated with a wide spectrum of disease. An Important part of this spectrum is caused by mutations of mitochondrial DNA (mtDNA). These class I OXPHOS diseases are covered in part I of this two-part review. Dysfunction of mitochondrial OXPHOS has also emerged as an important component of a range of predominantly neurodegenerative diseases in which the mitochondrial abnormality is most probably secondary. These class II OXPHOS diseases are due to mutations of genes not encoding OXPHOS subunits or are caused by exogenous or endogenous OXPHOS toxins. Class II mitochondrial diseases and the mitochondrion's role in apoptosis are covered in part II (Lancet 2000; 355: 389 94). PMID- 10675087 TI - New light on cranial surgery in ancient Rome. PMID- 10675088 TI - Association studies of genetic polymorphisms and complex disease. PMID- 10675089 TI - Biocompatibility and acute renal failure. PMID- 10675090 TI - Biocompatibility and acute renal failure. PMID- 10675091 TI - Biocompatibility and acute renal failure. PMID- 10675092 TI - Biocompatibility and acute renal failure. PMID- 10675093 TI - Mini mental state examination. PMID- 10675094 TI - An unfortunate case of allergy to latex. PMID- 10675095 TI - Are drugs interchangeable? PMID- 10675096 TI - Are drugs interchangeable? PMID- 10675097 TI - Are drugs interchangeable? PMID- 10675098 TI - Pseudoseizures versus epileptic seizures in pregnancy. PMID- 10675099 TI - Cortical origins of pathological pain. PMID- 10675100 TI - Medical implications of HGP's sequence of chromosome 22. PMID- 10675101 TI - Treatment and secondary prevention of stroke. PMID- 10675102 TI - Treatment and secondary prevention of stroke. PMID- 10675103 TI - Treatment and secondary prevention of stroke. PMID- 10675104 TI - Why such diverse prices of infant formula in Europe? PMID- 10675105 TI - Youthful memories: a tribute to David Baum. PMID- 10675106 TI - Baby 1, obstetrician 0. PMID- 10675107 TI - Overoptimism about cancer. PMID- 10675108 TI - Improving outcome of cataract surgery in developing countries. PMID- 10675109 TI - Measures and meaning of blood pressure. PMID- 10675110 TI - Response of metastatic breast cancer to trastuzumab? PMID- 10675111 TI - Should men still go bald gracefully? PMID- 10675112 TI - Progressive forms of MS: classification streamlined or consensus overturned? PMID- 10675113 TI - Lack of significant hormonal effects and controlled trials of phyto-oestrogens. PMID- 10675114 TI - Risks of leukaemia and solid tumours in individuals with Down's syndrome. AB - BACKGROUND: Individuals with Down's syndrome have a greater risk of leukaemia than the general population, but reliable estimates of the age-specific risk are lacking and little is known about the risk of solid tumours. METHODS: We identified 2814 individuals with Down's syndrome from the Danish Cytogenetic Register, and linked the data to the Danish Cancer Registry. The number of person years at risk was 48453. Standardised incidence ratio (SIR) and 95% CI were calculated of the basis of cancer rates specific for age and sex in the general population. FINDINGS: 60 cases of cancer were found, with 49.8 expected (SIR 1.20 [95% CI 0.92-1.55]). Leukaemia constituted 60% of cases of malignant disease overall and 97% of cases in children. The SIR for leukaemia varied with age, being 56 (38-81) at age 0-4 years and 10 (4-20) at 5-29 years. No cases of leukaemia were seen after the age of 29 years. The SIR for acute myeloid leukaemia was 3.8 (1.7-8.4) times higher than that for acute lymphoblastic leukaemia in children aged 0-4 years. The cumulative risk for leukaemia by the age of 5 years was 2.1% and that by 30 years was 2.7%. Only 24 solid tumours were seen, with 47.8 expected (0.50 [0.32-0.75]). No cases of breast cancer were found, with 7.3 expected (p=0.0007). Higher than expected numbers of testicular cancers, ovarian cancers, and retinoblastomas were seen but were not significant. INTREPRETATION: The occurrence of cancer in Down's syndrome is unique with a high risk of leukaemia in children and a decreased risk of solid tumours in all age groups. The distinctive pattern of malignant diseases may provide clues in the search for leukaemogenic genes and tumour-suppressor genes on chromosome 21. PMID- 10675115 TI - Laparoscopic or conventional Nissen fundoplication for gastro-oesophageal reflux disease: randomised clinical trial. The Netherlands Antireflux Surgery Study Group. AB - BACKGROUND: For the surgical treatment of gastrooesophageal reflux disease (GORD), laparoscopic Nissen fundoplication has largely replaced the open procedure. Retrospective and prospective non-randomised studies have shown similar results after laparoscopic Nissen fundoplication compared with the open procedure. METHODS: In a multicentre randomised trial candidates for surgical treatment of GORD were randomly assigned to either laparoscopic or open 360 degrees Nissen fundoplication. Primary endpoints were dysphagia, recurrent GORD, and intrathoracic hernia. Secondary endpoints were effectiveness and quality of life. This planned interim analysis focuses on endpoints and complications and in hospital costs. FINDINGS: At the time of interim analysis, 11 patients in the laparoscopic group and one in the conventional group had reached a primary endpoint (p=0.01; relative risk=8.8, 95% CI 1.2-66.3). This difference was caused mainly by whether or not patients had dysphagia (seven patients in the laparoscopic group and none in the conventional group, p=0.016). INTERPRETATION: Although laparoscopic Nissen fundoplication was as effective as the open procedure in controlling reflux, the significantly higher risk of reaching a primary endpoint in the laparoscopic group led us to stop the study. PMID- 10675116 TI - Systolic blood pressure and mortality. AB - BACKGROUND: The current systolic blood-pressure threshold for hypertension treatment is 140 mm Hg for all adults. WHO and the International Society of Hypertension have proposed that normal pressure be lower than 130 mm Hg, with an optimum pressure of less than 120 mm Hg. These recommendations are based largely on the assumption that cardiovascular and overall mortality depend in a strictly increasing manner on systolic blood pressure. The Framingham study was instrumental in establishing this viewpoint. We reassessed data from that study to find out whether the relation is strictly increasing or whether there is a threshold in this relation. METHODS: We used logistic splines to model the relation of risk of cardiovascular and all-cause death with systolic blood pressure, using age-specific and sex-specific rates. We tested for the independence of the slope parameters from age and sex, and the reduced model with common slopes was used to produce a model different from the conventional linear logistic model. FINDINGS: Against the predictions of the linear logistic model, neither all-cause nor cardiovascular deaths depended on systolic blood pressure in a strictly increasing manner. The linear logistic model was rejected by the Framingham data. Instead, risk was independent of systolic blood pressure for all pressures lower than a threshold at the 70th percentile for a person of a given age and sex. Risk sharply increased with pressure higher than the 80th percentile. Since systolic blood pressure steadily increases with age, the threshold increases with age, but more rapidly in women than in men. INTERPRETATION: The Framingham data contradict the concept that lower pressures imply lower risk and the idea that 140 mm Hg is a useful cut-off value for hypertension for all adults. There is an age-dependent and sex-dependent threshold for hypertension. A substantial proportion of the population who would currently be thought to be at increased risk are, therefore, at no increased risk. PMID- 10675117 TI - Cost-effectiveness of public-funded options for cataract surgery in Mysore, India. AB - BACKGROUND: In India 3.8 million people become blind due to cataracts every year. We assessed the cost-effectiveness of public-funded options for delivering cataract surgery in Mysore, Karnataka State, India. METHODS: Three types of delivery of cataract surgery were studied: mobile government camps, walk-in services at a state medical college hospital, and patients transported in from satellite clinics to a non-governmental hospital. We assessed outcomes in a systematic sample of patients operated on in 1996-97 by follow-up at home; average costs by provider derived from actual expenditures during the year. FINDINGS: Almost half the patients operated on in government camps were dissatisfied with the outcome (34/70, 49% [95% CI 36-61]). More than one third were blind in the operated eye (25/70, 36% [25-48]). User satisfaction was higher with other providers (medical college hospital 82% [63-94]; non-government hospital 85% [72-93]), and fewer patients remained blind. Camps were a low-cost option, but the poor outcomes reduced their cost-effectiveness to US$97 per patient. The state medical college hospital was least cost-effective, at US$176 per patient, and the non-governmental hospital was the most cost-effective at US$54 per patient. INTERPRETATION: The government of India should review its policy for government camp surgery, and consider alternatives, such as transporting patients to better permanent facilities. India and other developing countries should monitor outcomes in cataract surgery programmes, as well as throughput. PMID- 10675118 TI - Effects of a clinical-practice guideline and practice-based education on detection and outcome of depression in primary care: Hampshire Depression Project randomised controlled trial. AB - BACKGROUND: Depression is a major individual and public-health burden throughout the world and is managed mainly in primary care. The most effective strategy to reduce this burden has been believed to be education of primary-care practitioners. We tested this assumption by assessing the effectiveness of an educational programme based on a clinical-practice guideline in improving the recognition and outcome of primary-care depression. METHODS: We carried out a randomised controlled trial in a representative sample of 60 primary-care practices (26% of the total) in an English health district. Education was delivered to practice teams and quality tested by feedback from participants and expert raters. The primary endpoints were recognition of depression, defined by the hospital anxiety and depression (HAD) scale, and clinical improvement. Analysis was by intention to treat. FINDINGS: The education was well received by participants, 80% of whom thought it would change their management of patients with depression. 21409 patients were screened, of whom 4192 were classified as depressed by the HAD scale. The sensitivity of physicians to depressive symptoms was 39% in the intervention group and 36% in the control group after education (odds ratio 1.2 [95% CI 0.88-1.61]). The outcome of depressed patients as a whole at 6 weeks or 6 months after the assessment did not significantly improve. INTERPRETATION: Although well received, this in-practice programme, which was designed to convey the current consensus on best practice for the care of depression, did not deliver improvements in recognition of or recovery from depression. PMID- 10675119 TI - Reversion of prion protein conformational changes by synthetic beta-sheet breaker peptides. AB - BACKGROUND: Transmissible spongiform encephalopathies are associated with a structural transition in the prion protein that results in the conversion of the physiological PrPc to pathological PrP(Sc). We investigated whether this conformational transition can be inhibited and reversed by peptides homologous to the PrP fragments implicated in the abnormal folding, which contain specific residues acting as beta-sheet blockers (beta-sheet breaker peptides). METHODS: We studied the effect of a 13-residue beta-sheet breaker peptide (iPrP13) on the reversion of the abnormal structure and properties of PrP(Sc) purified from the brains of mice with experimental scrapie and from human beings affected by sporadic and variant Creutzfeldt-Jakob disease. In a cellular model of familial prion disease, we studied the effect of the peptide in the production of the abnormal form of PrP in intact cells. The influence of the peptide on prion infectivity was studied in vivo by incubation time assays in mice with experimental scrapie. FINDINGS: The beta-sheet breaker peptide partly reversed in vitro PrP(Sc) to a biochemical and structural state similar to that of PrPc. The effect of the peptide was also detected in intact cells. Treatment of prion infectious material with iPrP13 delayed the appearance of clinical symptoms and decreased infectivity by 90-95% in mice with experimental scrapie. INTERPRETATION: Beta-sheet breaker peptides reverse PrP conformational changes implicated in the pathogenesis of spongiform encephalopathies. These peptides or their derivatives provide a useful tool to study the role of PrP conformation and might represent a novel therapeutic approach for prion-related disorders. PMID- 10675120 TI - Fever and leucopenia with steroids. PMID- 10675121 TI - Association between violence, psychosis, and relationship to victim in stalkers. AB - 50 stalkers were assessed before their trials. Serious violence was significantly associated with previous sexual intimacy between stalker and victim; such stalkers were significantly less likely than those who stalked strangers to have psychotic illness. PMID- 10675122 TI - Failure of elimination of paternal mitochondrial DNA in abnormal embryos. AB - Paternal mitochondrial DNA is normally eliminated from mammalian embryos. We have shown the presence of paternal mtDNA at the blastocyst stage in some abnormal human embryos. PMID- 10675123 TI - Postural dependency of the cerebral venous outflow. AB - We have shown that predominance of the jugular veins in cerebrovenous drainage is limited to the supine position. In the erect position, the vertebral venous system represents the major outflow pathway. PMID- 10675124 TI - Measles in a Dutch hospital introduced by an immuno-compromised infant from Indonesia infected with a new virus genotype. AB - A fatal measles case in an immunocompromised Indonesian child was associated with nosocomial transmission to health care workers. The virus isolated proved to represent a new genotype within clade G. PMID- 10675125 TI - Difference in nature of ruptured and unruptured cerebral aneurysms. AB - Although small aneurysms have an extremely low probability of rupture, most aneurysms that rupture are found to be less than 10 mm in diameter. Histological study of aneurysms and epidemiological analysis of subarachnoid haemorrhage revealed that ruptured and unruptured aneurysms are of a different nature. PMID- 10675126 TI - Sexual origins of placental dysfunction. AB - Severe placental dysfunction is much more common in pregnancies with a male than with a female fetus. Furthermore, the birthweight/placental weight ratio is increased in these pregnancies, consistent with fetal growth restriction, and is higher with a male fetus than with a female fetus. These observations of placental insufficiency may underlie the increased in-utero loss rate of male fetuses. PMID- 10675127 TI - Diphtheria in urban slums in north India. AB - We recorded a reappearance of cases of microbiologically confirmed diphtheria in a tertiary care hospital in north India. Poor immunisation coverage, population migrations, and overcrowded urban slums may be contributory factors. PMID- 10675128 TI - Embryo splitting produces primate "clone" PMID- 10675129 TI - Osteoarthritis research: on the verge of a revolution? PMID- 10675130 TI - Japan puts the rights of the disabled into the spotlight. PMID- 10675131 TI - Agency warns of crisis in beleaguered Democratic Republic of Congo. PMID- 10675132 TI - USA spearheads renewed efforts to combat AIDS. PMID- 10675133 TI - Cardiovascular gene therapy. AB - Vascular gene transfer potentially offers new treatments for cardiovascular diseases. It can be used to overexpress therapeutically important proteins and correct genetic defects, and to test experimentally the effects of various genes in a local vascular compartment. Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) gene transfers have improved blood flow and collateral development in ischaemic limb and myocardium. Promising therapeutic effects have been obtained in animal models of restenosis or vein-graft thickening with the transfer of genes coding for VEGF, nitric-oxide synthase, thymidine kinase, retinoblastoma, growth arrest homoeobox, tissue inhibitor of metalloproteinases, cyclin or cyclin-dependent kinase inhibitors, fas ligand and hirudin, and antisense oligonucleotides against transcription factors or cell cycle regulatory proteins. First experiences of VEGF gene transfer and decoy oligonucleotides in human beings have been reported. However, further developments in gene-transfer vectors, gene-delivery techniques and identification of effective treatment genes will be required before the full therapeutic potential of gene therapy in cardiovascular disease can be assessed. PMID- 10675134 TI - Death by decree. PMID- 10675135 TI - Mild cognitive impairment: conceptual basis and current nosological status. PMID- 10675136 TI - Minor-injury care by nurse practitioners or junior doctors. PMID- 10675137 TI - Minor-injury care by nurse practitioners or junior doctors. PMID- 10675138 TI - Minor-injury care by nurse practitioners or junior doctors. PMID- 10675139 TI - Minor-injury care by nurse practitioners or junior doctors. PMID- 10675140 TI - Minor-injury care by nurse practitioners or junior doctors. PMID- 10675141 TI - Glaucoma and paclitaxel. PMID- 10675142 TI - Screening for congenital dislocation of the hip. PMID- 10675143 TI - Screening for congenital dislocation of the hip. PMID- 10675144 TI - Acute vestibulopathy. PMID- 10675145 TI - Chronic pain. PMID- 10675146 TI - Y chromosome. PMID- 10675147 TI - Lowering of LDL cholesterol. PMID- 10675148 TI - Prophylaxis against respiratory syncytial virus in premature infants. PMID- 10675149 TI - Chagas' disease challenge. PMID- 10675150 TI - Neutral protamine Hagedorn insulin. PMID- 10675151 TI - Renal effects of cyclo-oxygenase-type-2 inhibition. PMID- 10675152 TI - Self-use of rapid tests for malaria diagnosis. PMID- 10675153 TI - Self-use of rapid tests for malaria diagnosis. PMID- 10675154 TI - Extreme potency of botulinum toxin. PMID- 10675155 TI - Tobacco and health-care professionals in Japan. PMID- 10675156 TI - Pathological laughing and crying. PMID- 10675157 TI - Ethics of intensive neonatal care. PMID- 10675158 TI - Assessment of nationwide cancer-screening programmes. PMID- 10675159 TI - Maternal blood pressure and birthweight. PMID- 10675160 TI - Development of practice guidelines. PMID- 10675161 TI - Should staff in long-stay hospitals for elderly patients be vaccinated against influenza? PMID- 10675162 TI - Might olfactory dysfunction be a marker of early Alzheimer's disease? PMID- 10675163 TI - Strategy after diagnosis of pancreatic dysplasia. PMID- 10675164 TI - Fall in mean arterial pressure and fetal growth restriction in pregnancy hypertension: a meta-analysis. AB - BACKGROUND: We investigated the relation between fetoplacental growth and the use of oral antihypertensive medication to treat mild-to-moderate pregnancy hypertension. METHODS: The study design was a metaregression analysis of published data from randomised controlled trials. Data from a paper that was regarded as an extreme statistical outliner were excluded from primary analyses. The change in (group) mean arterial pressure (MAP) from enrolment to delivery was compared with indicators of fetoplacental growth. FINDINGS: Greater mean difference in MAP with antihypertensive therapy was associated with the birth of a higher proportion of small-for-gestational-age (SGA) infants (slope: 0.09 [SD 0.03], r2=0.48, p=0.006, 14 trials) and lower mean birthweight significant after exclusion of data from another paper regarded as an extreme statistical outliner (slope: -14.49 [6.98] r=0.16, p=0.049, 27). No relation with mean placental weight was seen (slope -2.01 [1.62], r2=0.15, p=0.25, 11 trials). INTERPRETATION: Treatment-induced falls in maternal blood pressure may adversely affect fetal growth. Given the small maternal benefits that are likely to be derived from therapy, new data are urgently needed to elucidate the relative maternal and fetal benefits and risks of oral antihypertensive drug treatment of mild-to moderate pregnancy hypertension. PMID- 10675165 TI - Effects of influenza vaccination of health-care workers on mortality of elderly people in long-term care: a randomised controlled trial. AB - BACKGROUND: Vaccination of health-care workers has been claimed to prevent nosocomial influenza infection of elderly patients in long-term care. Data are, however, limited on this strategy. We aimed to find out whether vaccination of health-care workers lowers mortality and the frequency of virologically proven influenza in such patients. METHODS: In a parallel-group study, health-care workers in 20 long-term elderly-care hospitals (range 44-105 patients) were randomly offered or not offered influenza vaccine (cluster randomisation, stratified for policy for vaccination of patients and hospital size). All deaths among patients were recorded over 6 months in the winter of 1996-97. We selected a random sample of 50% of patients for virological surveillance for influenza, with combined nasal and throat swabs taken every 2 weeks during the epidemic period. Swabs were tested by tissue culture and PCR for influenza viruses A and B. FINDINGS: Influenza vaccine uptake in health-care workers was 50.9% in hospitals in which they were routinely offered vaccine, compared with 4.9% in those in which they were not. The uncorrected rate of mortality in patients was 102 (13.6%) of 749 in vaccine hospitals compared with 154 (22.4%) of 688 in no vaccine hospitals (odds ratio 0.58 [95% CI 0.40-0.84], p=0.014). The two groups did not differ for proportions of patients positive for influenza infection (5.4% and 6.7%, respectively); at necropsy, PCR was positive in none of 17 patients from vaccine hospitals and six (20%) of 30 from no-vaccine hospitals (p=0.055). INTERPRETATION: Vaccination of health-care workers was associated with a substantial decrease in mortality among patients. However, virological surveillance showed no associated decrease in non-fatal influenza infection in patients. PMID- 10675166 TI - Effect of pelvic-floor re-education on duration and degree of incontinence after radical prostatectomy: a randomised controlled trial. AB - BACKGROUND: Urinary incontinence is a common long-term complication after radical prostatectomy. Spontaneous recovery of normal urinary control after surgery can take 1-2 years. We aimed to investigate whether there was any beneficial effect of pelvic-floor re-education for patients with urinary incontinence as a result of radical prostatectomy. METHODS: 102 consecutive incontinent patients who had had radical retropubic prostatectomy for clinically localised prostate cancer and who could comply with the ambulatory treatment schedule in our hospital were randomised, after catheter removal, into a treatment group (n=50) and a control group (n=52). Patients in the treatment group took part in a pelvic-floor re education programme for as long as they were incontinent, and for a maximum of 1 year. The control group received placebo therapy. The primary endpoint was continence rate at 3 months. Incontinence was assessed objectively with the 1 h and 24 h pad tests and subjectively by the visual analogue scale. The groups were analysed on an intention-to-treat basis by ANOVA and chi2-test. FINDINGS: In the treatment group continence was achieved after 3 months in 43 (88%) of 48 patients. In the control group, continence returned after 3 months in 29 (56%) of 52 patients. At 1 year, the difference in proportion between treatment and control group was 14% (95% CI 2-27). In the treatment group improvement in both duration (log-rank test, p=0.0001) and degree of incontinence (Wald test, p=0.0010) was significantly better than in the control group. INTERPRETATION: Pelvic-floor re-education should be considered as a first-line option in curing incontinence after radical prostatectomy. PMID- 10675167 TI - Practice guidelines developed by specialty societies: the need for a critical appraisal. AB - BACKGROUND: There is increasing concern about the quality, reliability, and independence of practice guidelines. Because no information is available on the methodological quality of the guidelines developed by specialty societies, we undertook a survey on those published in peer-reviewed journals. METHODS: Practice guidelines produced by specialty societies and published in English between January, 1988, and July, 1998, where identified through MEDLINE. Their quality was assessed in terms of whether they reported: the type of professionals and stakeholders involved in the development process; the strategy to identify primary evidence; and an explicit grading of recommendations according to the quality of supporting evidence. FINDINGS: Overall, 431 guidelines were eligible for the study. Most did not meet the criteria: 67% did not report any description of the type of stakeholders, 88% gave no information on searches for published studies, and 82% did not give any explicit grading of the strength of recommendations. There was improvement over time for searches (from 2% to 18%, p<0.001) and explicit grading of evidence (from 6% to 27%, p<0.001). All three criteria for quality were met in only 22 (5%) guidelines. INTERPRETATION: Despite improvement over time, the quality of practice guidelines developed by specialty societies is unsatisfactory. Explicit methodological criteria for the production of guidelines shared among public agencies, scientific societies, and patients' associations need to be set up. Common standards of reporting, following the same principles that led to the CONSORT statement for randomised clinical trials, should be promoted. PMID- 10675168 TI - Classification of thyroid size by palpation and ultrasonography in field surveys. AB - BACKGROUND: Goitre surveys are used to assess the degree of iodine deficiency in a population. The change of goitre classification made by WHO in 1994 implied that a smaller thyroid size should be regarded as goitre. Furthermore, the acceptable goitre prevalence was lowered from 10% to 5%, and ultrasonography was recommended as a more precise method for diagnosis of goitre. We studied the effects of the change of palpation system, and compared the precision of the old and new systems with that of ultrasonographic examination. METHODS: We studied 225 schoolchildren (aged 7-14 years) in a highland village in Tanzania. The size of the thyroid was assessed in duplicate by ultrasonography and by WHO's 1960 and 1994 palpation systems. The latter were done by three examiners. Variations within and between examination methods and examiners were assessed, and measurement errors by ultrasonography were assessed from duplicate examinations. The sensitivity and specificity of the two palpation systems were calculated, with diagnosis by ultrasonography as the gold standard. Apparent palpation prevalences were calculated at a "true" 5% prevalence. FINDINGS: The lowered criterion for goitre resulted in an extra 20-33% of children being diagnosed as having goitre by palpation. The variation between repeat examinations was only slightly smaller by ultrasonography (kappa=0.63) than by experienced examiners (kappa=0.57-0.58). The variation between thyroid volume estimation by ultrasonography and the true volume was about 50% due to both measurement error and variation in the shape of thyroid lobes. The new goitre criterion decreased specificity from 76% to 29%, whereas sensitivity rose from 56% to 80%. In contrast, a suggested sharpening of the old criterion increased specificity to 90%. INTERPRETATION: A return to the old (1960) palpation criterion for goitre: "lobes larger than the terminal phalanxes of thumbs" and to an accepted palpation goitre prevalence of 10% can allow affordable monitoring of thyroid size through palpation in field surveys. PMID- 10675169 TI - Epidemiological and diagnostic aspects of the outbreak of pneumonic plague in Madagascar. AB - BACKGROUND: Plague is a re-emerging disease and pneumonic plague is the most feared clinical form. We describe a well-documented outbreak of pneumonic plague in Madagascar. METHODS: Field epidemiological data were collected. Biological tests (microscopy, culture of Yersinia pestis, F1 antigen ELISA and dipstick assays, IgG anti-F1 ELISA) were done on sputum, serum, or necropsy samples. The infection rate among 154 contacts was assessed by anti-F1 serological techniques. FINDINGS: The index case was a bubonic patient with a secondary lung infection, who contaminated a traditional healer and his family. Funeral ceremonies and attendance on patients contaminated other villagers. In total 18 cases were recorded, and eight died. F1 antigen could be detected in sputum by ELISA and dipstick tests as early as the second day after the onset of the symptoms and also 48 h after treatment. Among the contact population 13 of 154 (8.4%) have been exposed to the plague bacillus (symptomless or latent infections). INTERPRETATION: The F1 dipstick assay on sputum is an invaluable diagnostic tool for pneumonic plague. Treatment of patients and chemoprophylaxis of contacts were efficient in stopping the epidemic. PMID- 10675170 TI - A cause of pre-eclampsia? PMID- 10675171 TI - Identification by positron emission tomography of neuronal loss in acute vegetative state. AB - Positron emission tomography with the benzodiazepine receptor ligand carbon-11 labelled flumazenil Identified the extent of neuronal damage in acute vegetative state and predicted the possibility of recovery of consciousness and function. PMID- 10675172 TI - Three-dimensional ultrasonography: new prospects for ultrasound imaging of bone. AB - Radiography is used for the initial evaluation of suspected bone lesions. We have shown that surface images of bone can be obtained by three-dimensional ultrasonography. PMID- 10675173 TI - Neutropenia among survivors of atomic bomb explosion. AB - High incidence of neutropenia was found among atomic bomb survivors. We showed that the cause of neutropenia was due to NK or NK-like T-cell proliferative disorders, diagnosed by immunophenotypic, functional, and DNA analysis. PMID- 10675174 TI - Community-based study of genital schistosomiasis in men from Madagascar. AB - Detection of Schistosoma haematoblum eggs in 43% of semen samples with Increased levels of eosinophil cationic protein suggests that the genital organs of men are frequently affected with schistosomiasis. PMID- 10675176 TI - Mode of delivery and subsequent stress response. AB - We have shown that a baby's stress (saliva cortisol) and crying response to inoculation at 8 weeks was related to mode of delivery, with the greatest response shown in those born by assisted delivery and the least response in those born by elective caesarean section. PMID- 10675175 TI - Risk of congenital malformations associated with treatment of asthma during early pregnancy. AB - Studies assessing the risk of congenital malformations associated with the treatment of asthma during the first trimester of pregnancy are few, have limited power and support continuation of treatment. PMID- 10675178 TI - Drug-pricing "powder keg" stirs up US health-care politics. PMID- 10675177 TI - Work-related stress: can it be a thing of the past? PMID- 10675179 TI - Fresh allegations levelled at Toronto paediatrician. PMID- 10675180 TI - Concern grows about adolescent pregnancy in Cape Verde. PMID- 10675181 TI - Is screening for breast cancer with mammography justifiable? AB - BACKGROUND: A 1999 study found no decrease in breast-cancer mortality in Sweden, where screening has been recommended since 1985. We therefore reviewed the methodological quality of the mammography trials and an influential Swedish meta analysis, and did a meta-analysis ourselves. METHODS: We searched the Cochrane Library for trials and asked the investigators for further details. Meta-analyses were done with Review Manager (version 4.0). FINDINGS: Baseline imbalances were shown for six of the eight identified trials, and inconsistencies in the number of women randomised were found in four. The two adequately randomised trials found no effect of screening on breast-cancer mortality (pooled relative risk 1.04 [95% CI 0.84-1.27]) or on total mortality (0.99 [0.94-1.05]). The pooled relative risk for breast-cancer mortality for the other trials was 0.75 (0.67 0.83), which was significantly different (p=0.005) from that for the unbiased trials. The Swedish meta-analysis showed a decrease in breast-cancer mortality but also an increase in total mortality (1.06 [1.04-1.08]); this increase disappeared after adjustment for an imbalance in age. INTERPRETATION: Screening for breast cancer with mammography is unjustified. If the Swedish trials are judged to be unbiased, the data show that for every 1000 women screened biennially throughout 12 years, one breast-cancer death is avoided whereas the total number of deaths is increased by six. If the Swedish trials (apart from the Malmo trial) are judged to be biased, there is no reliable evidence that screening decreases breast-cancer mortality. PMID- 10675182 TI - Herb-drug interactions. AB - Concurrent use of herbs may mimic, magnify, or oppose the effect of drugs. Plausible cases of herb-drug interactions include: bleeding when warfarin is combined with ginkgo (Ginkgo biloba), garlic (Allium sativum), dong quai (Angelica sinensis), or danshen (Salvia miltiorrhiza); mild serotonin syndrome in patients who mix St John's wort (Hypericum perforatum) with serotonin-reuptake inhibitors; decreased bioavailability of digoxin, theophylline, cyclosporin, and phenprocoumon when these drugs are combined with St John's wort; induction of mania in depressed patients who mix antidepressants and Panax ginseng; exacerbation of extrapyramidal effects with neuroleptic drugs and betel nut (Areca catechu); increased risk of hypertension when tricyclic antidepressants are combined with yohimbine (Pausinystalia yohimbe); potentiation of oral and topical corticosteroids by liquorice (Glycyrrhiza glabra); decreased blood concentrations of prednisolone when taken with the Chinese herbal product xaio chai hu tang (sho-salko-to); and decreased concentrations of phenytoin when combined with the Ayurvedic syrup shankhapushpi. Anthranoid-containing plants (including senna [Cassia senna] and cascara [Rhamnus purshiana]) and soluble fibres (including guar gum and psyllium) can decrease the absorption of drugs. Many reports of herb-drug interactions are sketchy and lack laboratory analysis of suspect preparations. Health-care practitioners should caution patients against mixing herbs and pharmaceutical drugs. PMID- 10675183 TI - On the other side of the tracks. PMID- 10675184 TI - Unrecognised Mycobacterium tuberculosis. PMID- 10675185 TI - Unrecognised Mycobacterium tuberculosis. PMID- 10675186 TI - Unrecognised Mycobacterium tuberculosis. PMID- 10675187 TI - Unrecognised Mycobacterium tuberculosis. PMID- 10675188 TI - Contact-lens-associated microbial keratitis in The Netherlands and Scotland. PMID- 10675189 TI - Immunohistochemical detection of lymph-node metastases. PMID- 10675190 TI - Immunohistochemical detection of lymph-node metastases. PMID- 10675191 TI - Soluble thrombomodulin in hypercholesterolaemic patients. PMID- 10675192 TI - Adenovirus in EV71-associated hand, foot, and mouth disease. PMID- 10675193 TI - Adenovirus in EV71-associated hand, foot, and mouth disease. PMID- 10675194 TI - Cluster headache and melatonin. PMID- 10675195 TI - Folate supplementation and neural-tube defects. PMID- 10675196 TI - Insulin-like growth factor I for growth hormone therapy. PMID- 10675197 TI - Oral cobalamin therapy. PMID- 10675198 TI - After postmodernism. PMID- 10675199 TI - After postmodernism. PMID- 10675200 TI - European role in global cardiology. PMID- 10675201 TI - Game plan for cancer care. PMID- 10675202 TI - John Lykoudis and peptic ulcer disease. PMID- 10675204 TI - Vasculitic neuropathies. PMID- 10675203 TI - The role of the delayed rectifier component IKs in dog ventricular muscle and Purkinje fibre repolarization. AB - 1. The relative contributions of the rapid and slow components of the delayed rectifier potassium current (IKr and IKs, respectively) to dog cardiac action potential configuration were compared in ventricular myocytes and in multicellular right ventricular papillary muscle and Purkinje fibre preparations. Whole-cell patch-clamp techniques, conventional microelectrode and in vivo ECG measurements were made at 37C. 2. Action potential duration (APD) was minimally increased (less than 7%) by chromanol 293B (10 microM) and L-735,821 (100 nM), selective blockers of IKs, over a range of pacing cycle lengths (300-5000 ms) in both dog right ventricular papillary muscles and Purkinje fibre strands. D Sotalol (30 microM) and E-4031 (1 microM), selective blockers of IKr, in the same preparations markedly (20-80%) lengthened APD in a reverse frequency-dependent manner. 3. In vivo ECG recordings in intact anaesthetized dogs indicated no significant chromanol 293B (1 mg kg-1 i.v.) effect on the QTc interval (332.9 +/- 16.1 ms before versus 330.5 +/- 11.2 ms, n = 6, after chromanol 293B), while D sotalol (1 mg kg-1 i.v.) significantly increased the QTc interval (323.9 +/- 7.3 ms before versus 346.5 +/- 6.4 ms, n = 5, after D-sotalol, P < 0.05). 4. The current density estimated during the normal ventricular muscle action potential (i.e. after a 200 ms square pulse to +30 mV or during a 250 ms long 'action potential-like' test pulse) indicates that substantially more current is conducted through IKr channels than through IKs channels. However, if the duration of the square test pulse or the 'action potential-like' test pulse was lengthened to 500 ms the relative contribution of IKs significantly increased. 5. When APD was pharmacologically prolonged in papillary muscle (1 microM E-4031 and 1 microg ml-1 veratrine), 100 nM L-735,821 and 10 microM chromanol 293B lengthened repolarization substantially by 14.4 +/- 3.4 and 18. 0 +/- 3.4% (n = 8), respectively. 6. We conclude that in this study IKs plays little role in normal dog ventricular muscle and Purkinje fibre action potential repolarization and that IKr is the major source of outward current responsible for initiation of final action potential repolarization. Thus, when APD is abnormally increased, the role of IKs in final repolarization increases to provide an important safety mechanism that reduces arrhythmia risk. PMID- 10675205 TI - Responsiveness of outcome measures in randomised controlled trials in neurology. PMID- 10675206 TI - Corticobasal degeneration. PMID- 10675207 TI - Pattern of dopaminergic loss in the striatum of humans with parkinsonism induced by MPTP. PMID- 10675208 TI - Neurological emergencies: acute stroke. AB - Stroke causes a vast amount of death and disability throughout the world, yet for many healthcare professionals it remains an area of therapeutic nihilism, and thus uninteresting. This negative perception is shared by the general public, who often have a poor understanding of the early symptoms and significance of a stroke. Yet within the past few years there have been many important developments in the approach to caring for stroke patients, for both the acute management and secondary prevention. After the completion of numerous clinical trials, there is now robust evidence to either support or discredit various interventions. Even more exciting is the prospect of yet more data becoming available in the near future, testing a whole array of treatments, as clinical interest in stroke expands exponentially. In this review an evidence based approach to the management of acute stroke within the first few days is presented, including ischaemic and haemorrhagic events, but not subarachnoid haemorrhage. It is explained why stroke is regarded as a medical emergency, and the importance of a rational, methodic approach to the initial assessment, which is the key to accurate diagnosis and subsequent management, is emphasised. The potential early problems associated with stroke are identified and specific interventions for different stroke types are discussed. The review ends with a brief discussion of the implications that the evolving treatments have for the organisation of modern stroke services. PMID- 10675209 TI - Tuberculous meningitis. PMID- 10675210 TI - Role of clinical, radiological, and neurophysiological changes in predicting the outcome of tuberculous meningitis: a multivariable analysis. AB - OBJECTIVES: The role of EEG and evoked potentials has not been evaluated in predicting the prognosis of tuberculous (TB) meningitis. The present study was aimed at evaluating the prognostic significance of clinical, radiological, and neurophysiological variables using multi-variable analysis. METHODS: Patients with TB meningitis diagnosed on the basis of clinical, radiological, and CSF criteria have been prospectively evaluated. All the patients were subjected to a detailed neurological evaluation. The outcome was defined 6 months after starting treatment on the basis of the Barthel index (BI) score into poor (BI <12) and good recovery (BI> or =12). Death was included in the poor recovery group for statistical analysis. Thirteen clinical (age, sex, seizure, focal weakness, stage of meningitis, Glasgow coma scale score, methyl prednisolone therapy), CT (infarction, hydrocephalus, tuberculoma) and neurophysiological (EEG, motor and somatosensory evoked potentials) variables were evaluated employing single variable logistic regression followed by multivariable logistic regression analysis. The best set of predictors were obtained by stepdown logistic regression analysis. RESULTS: Fifty four patients were included in the present study. Their age ranged between 5 and 62 years, 11 were children younger than 12 years and 14 were female. Nine patients were in stage I meningitis, 12 in stage II, and 33 in stage III. On single variable logistic regression analysis the significant predictors of 6 months outcome of TB meningitis included focal weakness, Glasgow coma scale (GCS), motor evoked potential (MEP) and somatosensory evoked potential (SEP). On multivariable analysis the best set of predictors comprised focal weakness, GCS, and SEP. CONCLUSIONS: In patients with TB meningitis focal weakness, GCS, and SEP are the best predictors of 6 month outcome. PMID- 10675211 TI - Corticobasal ganglionic degeneration and/or frontotemporal dementia? A report of two overlap cases and review of literature. AB - OBJECTIVE: According to the existing viewpoint, Corticobasal degeneration (CBD) is thought of as a predominantly extrapyramidal motor disorder that is distinct and unrelated to frontotemporal dementia (FTD), the most common form of non Alzheimer dementias. A lack of understanding of the aetiopathogenesis, and poor correlation between the pathology and the clinical syndromes, has resulted in a disparity in the classification of cases of non-Alzheimer dementias. This report intends to highlight the overlap between FTD and CBD in the light of the evolution of these terms, and to discuss the implications of these findings on the nosology of CBD and the classification of non-Alzheimer dementias. METHODS AND RESULTS: Two cases who presented with cognitive dysfunction, which, on comprehensive neuropsychological testing warranted an antemortem diagnosis of FTD are reported. A detailed necropsy study of their brains, however, favoured a pathological diagnosis of CBD. The literature on the overlap between CBD and FTD is also reviewed. CONCLUSIONS: Firstly, evidence is emerging to suggest that the clear distinction drawn between FTD and CBD by the existing viewpoint, needs revision. Secondly, until such time that a comprehensive classification of non Alzheimer dementias is evolved, it may be better to distinguish between the clinical and pathological levels of description and to classify cases, in vivo, on the basis of the clinical phenotype. PMID- 10675212 TI - Pattern of dopaminergic loss in the striatum of humans with MPTP induced parkinsonism. AB - OBJECTIVES: To examine the distribution of striatal dopaminergic function in humans with parkinsonism induced by 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine (MPTP) to determine if there is a caudate-putamen gradient as is seen in idiopathic Parkinson's disease. METHODS: We scanned nine humans exposed to MPTP with parkinsonism ranging from minimal to severe using [(18)F]fluorodopa (FD) and high resolution PET. The results were compared with those of 10 patients with Parkinson's disease and six normal subjects. RESULTS: In the MPTP group there was an equal degree of reduction of dopaminergic function in the caudate and putamen. This was different from the greater putaminal than caudate loss in Parkinson's disease (p<0.001). CONCLUSIONS: Parkinson's disease is not caused by transient exposure to MPTP. PMID- 10675213 TI - Atrophy of the corpus callosum associated with a decrease in cortical benzodiazepine receptor in large cerebral arterial occlusive diseases. AB - OBJECTIVES: It remains controversial whether selective neuronal ischaemic change develops in patients with occlusion of the large cerebral arteries. Previous studies have shown atrophy of the corpus callosum with reduced cortical oxygen metabolism in large cerebral arterial occlusive diseases, which might be indirect evidence of loss of the neurons in cortical layer 3. Recent studies of patients with ischaemic cerebrovascular diseases have demonstrated reduced central benzodiazepine receptor (BZR) binding in the normal appearing cortical areas, which might be more direct evidence of changes of the neurons. Although pathophysiology of the decreased BZR is unclear, a decrease in the cortical BZR binding with neuronal loss would cause atrophy of the corpus callosum. The purpose of this study was to determine whether atrophy of the corpus callosum is associated with a decrease in cortical BZR binding in large cerebral arterial occlusive diseases. METHODS: Seven patients with occlusive diseases of the middle cerebral or internal carotid artery and only minor subcortical infarctions were studied. Single photon emission tomographic images of (123)I labelled iomazenil (IMZ) obtained 180 minutes after injection were analysed for BZR binding. The midsagittal corpus callosum area/skull area ratio (on T1 weighted magnetic resonance images) was compared with the cerebral IMZ uptake/cerebellar IMZ uptake ratio. RESULTS: Compared with 23 age and sex matched control subjects, the patients had significantly decreased callosal area/skull area ratio. The degree of corpus callosum atrophy was significantly and strongly (rho=0.99, p<0.02) correlated with that of the decreases in the mean cerebral cortical IMZ uptake ratio. CONCLUSION: Corpus callosum atrophy may occur in association with a decrease in cortical BZR binding in large cerebral arterial occlusive diseases. Corpus callosum atrophy with decreased cortical BZR binding might reflect cortical neuronal damage in large cerebral arterial occlusive diseases. PMID- 10675214 TI - Value of somatosensory and motor evoked potentials in predicting arm recovery after a stroke. AB - OBJECTIVES: Prediction of motor recovery in the arm in patients with stroke is generally based on clinical examination. However, neurophysiological measures may also have a predictive value. The aims of this study were to assess the role of somatosensory (SSEPs) and motor (MEPs) evoked potentials in the prediction of arm motor recovery and to determine whether these measures added further predictive information to that gained from clinical examination. METHODS: Sixty four patients who had had a stroke and presented with obvious motor deficit of the arm were examined in terms of three clinical variables (motor performance, muscle tone, and overall disability) and for SSEPs and MEPs. Clinical and neurophysiological examinations were done at entry to the study (2 to 5 weeks poststroke), and at about 2 months after stroke. Further clinical follow up was conducted at 6 and 12 months after stroke. RESULTS: Neurophysiological measures made in the acute phase were of little use alone in predicting motor recovery of the arm at 2, 6, and 12 months after stroke. At 2 months, the absence of SSEPs and MEPs indicated a very poor outcome. Conversely, if the responses were preserved, a great variation in motor outcome was found. Multiple regression analysis showed that the addition of SSEPs and MEPs to the clinical examination increased the possibility of predicting arm recovery in the long term. In the acute phase, the combination of the motor score and SSEPs were best able to predict outcome. The long term outcome based on variables taken at 2 months, was best predicted through incorporating the three clinical measures and MEPs. CONCLUSIONS: Neurophysiological measures alone are of limited value in predicting long term outcome. However, predictive accuracy is substantially improved through the combined use of both of these measures and clinical variables. PMID- 10675215 TI - The notion of "warning leaks" in subarachnoid haemorrhage: are such patients in fact admitted with a rebleed? AB - OBJECTIVE: Often patients with subarachnoid haemorrhage (SAH) recall a recent episode of acute severe headache, usually interpreted as a "warning headache" or first SAH. An alternative explanation is recall bias. The clinical and radiological features of patients with SAH were studied in relation to previous headaches or later rebleeding. METHODS: Patients with either a previous headache episode or a subsequent rebleed were selected from the SAH database in Utrecht within 1 month of the index SAH. The clinical condition was graded on the World Federation of Neurological Surgeons (WFNS) scale. The CT was reviewed and the amounts of subarachnoid blood, hydrocephalus, and intraventricular, intracerebral, and subdural blood were rated. Proportions were compared by unpaired or paired t test. RESULTS: Forty four of 390 patients (11%) had had a severe headache before their index SAH (11 of these had a subsequent rebleed); 31 other patients had a rebleed in hospital but no preceding headache. Patients with and without preceding headache did not differ in level of consciousness (14 of 44 v 11 of 31 were comatose), nor in any of the radiological features. After rebleeding (42 patients), 37 of 42 patients were comatose (v 11 of 42 before), and CT showed higher proportions of intracerebral haemorrhage (17%), intraventricular haemorrhage, (27%), and hydrocephalus (12%) than baseline scans. Intraventricular haemorrhage was twice as frequent after rebleeding than at baseline. CONCLUSIONS: The clinical and radiological features of patients admitted with SAH after a preceding bout of headache did not differ from those without such an episode, and are clearly dissimilar from those after documented rebleeds. The findings challenge the existence of minor "warning headaches". PMID- 10675216 TI - Complications and outcome in patients with aneurysmal subarachnoid haemorrhage: a prospective hospital based cohort study in the Netherlands. AB - OBJECTIVE: The aim of this study was to investigate prospectively in an unselected series of patients with an aneurysmal subarachnoid haemorrhage what at present the complications are, what the outcome is, how many of these patients have "modern treatment"-that is, early obliteration of the aneurysm and treatment with calcium antagonists-what factors cause a delay in surgical or endovascular treatment, and what the estimated effect on outcome will be of improved treatment. METHODS: A prospective, observational cohort study of all patients with aneurysmal subarachnoid haemorrhage in the hospitals of a specified region in The Netherlands. The condition on admission, diagnostic procedures, and treatments were recorded. If a patient had a clinical deterioration, the change in Glasgow coma score (GCS), the presence of focal neurological signs, the results of additional investigations, and the final diagnosed cause of the deterioration were recorded. Clinical outcome was assessed with the Glasgow outcome scale (GOS) at 3 month follow up. In patients with poor outcome at follow up, the cause was diagnosed. RESULTS: Of the 110 patients, 47 (43%) had a poor outcome. Cerebral ischaemia, 31 patients (28%), was the most often occurring complication. Major causes of poor outcome were the effects of the initial haemorrhage and rebleeding in 34% and 30% of the patients with poor outcome respectively. Of all patients 102 (93%) were treated with calcium antagonists and 45 (41%) patients had early treatment to obliterate the aneurysm. The major causes of delay of treatment were a poor condition on admission or deterioration shortly after admission, in 31% and 23% respectively. CONCLUSIONS: In two thirds of the patients with poor outcome the causes of poor outcome are the effects of the initial bleeding and rebleeding. Improved treatment of delayed or postoperative ischaemia will have only minor effects on the outcome of patients with subarachnoid haemorrhage. PMID- 10675218 TI - Alzheimer's disease. PMID- 10675217 TI - Validity of carbohydrate deficient transferrin and other markers as diagnostic aids in the detection of alcohol related seizures. AB - OBJECTIVE: The role of alcohol misuse in the genesis of seizures is probably often undetected. The aim was to investigate the utility of carbohydrate deficient transferrin (CDT) compared with other biomarkers and clinical examination in the diagnosis of alcohol related seizures. METHODS: The study included consecutively 158 seizure patients-83 men and 75 women-with mean age 45 (16-79) years. Seizures related to alcohol use were identified by a score > or =8 in the alcohol use disorders identification test (AUDIT positive). AUDIT was applied as the gold standard to which sensitivity and specificity of the various markers were related. Blood samples were obtained from 150 patients on admission and analysed for ethanol, liver enzymes, and CDT, using AXIS Biochemicals' %CDT TIA kit. RESULTS: 53 patients (34%) were AUDIT positive. Using the commonly applied decision value for %CDT of 5.0%, a sensitivity of 41% and a specificity of 84% were obtained. Analysis of receiver operator characteristics (ROC) curves disclosed an optimal cut off value for %CDT of 5.4%, which yielded a sensitivity of 39% and a specificity of 88%. At a specificity of 80%, the sensitivity was 43% for %CDT and 26% for GGT. The %CDT sensitivity was markedly higher for men than for women. Compared with GGT, ASAT, ALAT, and ASAT/ALAT ratio, CDT was the best single biomarker for alcohol related seizures. However, even in the subgroup of withdrawal seizures, the sensitivity level barely exceeded 50%. Clinicians scored alcohol as the main cause of the seizure in only 19 cases (12%). In 38 (24%) cases, clinicians suspected that alcohol had a role (sensitivity of 62% at a specificity of 89%). Their ability to identify AUDIT positive patients was better than that of any biomarker, but many cases were missed. Agreement of clinicians' scores to CDT was only fair (kappa=0.28). CDT concentrations were significantly increased among alcohol abstaining patients on enzyme-inducing antiepileptic drugs. Six out of 16 patients with false positive CDT results were exposed to such drugs. CONCLUSIONS: CDT is not recommended as a stand alone marker for alcohol related seizures, but may provide a useful contribution to the overall diagnostic investigation of seizures. Confirmatory studies are needed as to the apparent vulnerability of CDT to antiepileptic drugs. PMID- 10675219 TI - Prevalence of multiple sclerosis in the L'Aquila district, central Italy. AB - OBJECTIVE: To estimate the prevalence of multiple sclerosis in the L'Aquila district, central Italy. METHODS: All available case sources were screened. Definite and probable cases of multiple sclerosis, classified according to the Poser criteria, were considered as prevalent cases. RESULTS: On the prevalence day, 31 December 1996, 158 patients (105 women and 53 men; ratio 2:1) affected by definite (n=131) or probable (n=27) multiple sclerosis were alive and resident in the L'Aquila district. Mean (SD) age was 38.4 (11.9) years (38.9 (11.7) years for women and 38.5 (12.3) years for men, p=0.9). The overall crude prevalence was 53.0/100 000 (95% confidence interval (95% CI)=45.4-62.0); 68.4/100 000 (95% CI=56. 5-82.8) in women, and 36.7/100 000 (95% CI=28.1-48.0) in men. The prevalence was similar (55.9/100 000) when standardised to the 1996 European population. Mean (SD) age at onset of multiple sclerosis was 29.4 (9.6) years and mean (SD) duration of the disease was 9.4 (7.4) years, without any significant difference between sexes. Mean age at onset was significantly higher in patients with the primary progressive than in those with the relapsing-remitting course (p=0. 0002, Scheffe's test). CONCLUSIONS: The prevalence found in the L'Aquila district gives support to the consideration of Italy as an area in which multiple sclerosis has been shown to have high prevalence at least in the populations that were surveyed recently. PMID- 10675220 TI - Evaluation of (18)fluorodeoxyglucose positron emission tomography ((18)FDG PET) in the detection of malignant peripheral nerve sheath tumours arising from within plexiform neurofibromas in neurofibromatosis 1. AB - OBJECTIVES: The ability of (18)fluorodeoxyglucose positron emission tomography ((18)FDG PET) to detect malignant change in plexiform neurofibromas from patients with neurofibromatosis 1 (NF1) was evaluated. METHODS: Eighteen NF1 patients who presented with pain, increase in size, or neurological deficit associated with a plexiform neurofibroma were assessed. Magnetic resonance imaging determined the site and extent of the lesion. Qualitative(18)FDG PET was performed and the standard uptake value (SUV) measured the regional glucose metabolism. Histological confirmation of the diagnosis was obtained in 10 patients. RESULTS: Twenty three plexiform neurofibromas were detected in 18 patients. Seven malignant peripheral nerve sheath tumours, four high grade and three low grade tumours, occurred in five patients. In one patient the clinical and radiological characteristics of the tumour suggested malignancy, but histology was inconclusive. Fifteen benign plexiform neurofibromas were identified in 12 patients and these findings were confirmed histologically in five lesions from four patients. Ten plexiform neurofibromas occurring in eight patients were considered benign on(18)FDG PET and the patients did not undergo surgery. They remained stable or their symptoms improved on clinical follow up (median 9 months). The results of qualitative (18)FDG PET were interpreted as indicating that 13 plexiform neurofibromas were benign and 10 were malignant. No malignant tumours were classified as benign, but two benign tumours were reported as malignant. The SUV was calculated for 20 tumours and was significantly higher in five malignant tumours 5.4 (SD 2.4), than in 15 benign tumours 1.54 (SD 0.7), p=0.002. There was an overlap between benign and malignant tumours in the SUV range 2.7-3.3. CONCLUSIONS: (18)FDG PET is helpful in determining malignant change in plexiform neurofibromas in NF1. Increased separation between benign and malignant lesions could be obtained by calculating the SUV at about 200 minutes after injection of (18)FDG, when the peak activity concentration is obtained in malignant tumours. PMID- 10675221 TI - Fear conditioned potentiation of the acoustic blink reflex in patients with cerebellar lesions. AB - OBJECTIVE: To investigate whether the human cerebellum takes part in fear conditioned potentiation of the acoustic blink reflex. METHODS: A group of 10 cerebellar patients (eight patients with lesions involving the medial cerebellum, two patients with circumscribed lesions of the cerebellar hemispheres) was compared with a group of 16 age and sex matched healthy control subjects. The fear conditioned potentiation paradigm consisted of three phases. During the first, habituation phase subjects received 20 successive acoustic blink stimuli. In the subsequent fear conditioning phase, subjects passed through 20 paired presentations of the unconditioned fear stimulus (US; an electric shock) and the conditioned stimulus (CS; a light). Thereafter, subjects underwent the potentiation phase, which consisted of a pseudorandom order of 12 trials of the acoustic blink stimulus alone, 12 acoustic blink stimuli paired with the conditioned stimulus, and six conditioned stimuli paired with the unconditioned stimulus. The EMG of the acoustic blink reflex was recorded at the orbicularis oculi muscles. The potentiation effect was determined as the difference in normalised peak amplitude of the blink reflex evoked by pairs of CS and acoustic blink stimuli and evoked by the acoustic stimulus alone. RESULTS: In the habituation phase, short term habituation of the acoustic blink reflex was preserved in all cerebellar patients. However, in the potentiation phase, the potentiation effect of the blink reflex was significantly reduced in patients with medial cerebellar lesions compared with the controls (mean (SD) potentiation effect (%), patients: -6.4 (15.3), controls: 21.6 (35.6)), but was within normal limits in the two patients with lateral lesions. CONCLUSIONS: The present findings suggest that the human medial cerebellum is involved in associative learning of non-specific aversive reactions-that is, the fear conditioned potentiation of the acoustic blink reflex. PMID- 10675222 TI - The London handicap scale: a re-evaluation of its validity using standard scoring and simple summation. AB - OBJECTIVE: To assess the validity of the London handicap scale (LHS) using a simple unweighted scoring system compared with traditional weighted scoring METHODS: 323 patients admitted to hospital with acute stroke were followed up by interview 6 months after their stroke as part of a trial looking at the impact of a family support organiser. Outcome measures included the six item LHS, the Dartmouth COOP charts, the Frenchay activities index, the Barthel index, and the hospital anxiety and depression scale. Patients' handicap score was calculated both using the standard procedure (with weighting) for the LHS, and using a simple summation procedure without weighting (U-LHS). Construct validity of both LHS and U-LHS was assessed by testing their correlations with the other outcome measures. RESULTS: Cronbach's alpha for the LHS was 0.83. The U-LHS was highly correlated with the LHS (r=0.98). Correlation of U-LHS with the other outcome measures gave very similar results to correlation of LHS with these measures. CONCLUSION: Simple summation scoring of the LHS does not lead to any change in the measurement properties of the instrument compared with standard weighted scoring. Unweighted scores are easier to calculate and interpret, so it is recommended that these are used. PMID- 10675223 TI - Upregulation of matrix metalloproteinase-9 in the cerebrospinal fluid of patients with acute Lyme neuroborreliosis. AB - It was investigated (1) whether metalloproteinase-9 (MMP-9), MMP-3, and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1, the natural tissue inhibitor of MMP-9) are increased in the CSF of patients with Lyme neuroborreliosis and (2) whether macrophages can express MMP-9 when stimulated with Borrelia burgdorferi. Zymography showed MMP-9 activity in 26 of 31 (84%) CSF samples from patients with acute stage 2 Lyme neuroborreliosis, but not in 20 controls with non-inflammatory neurological disorders. Activity of MMP-2 was detected in all CSF samples in both patients with neuroborreliosis and controls, suggesting a constitutive release of MMP-2. Using enzyme linked immunosorbent assay (ELISA) MMP-3 (which can activate MMP-9) was detected in low concentrations in the CSF of 13 of 29 patients with neuroborreliosis, but not in controls. TIMP-1 was increased twofold in CSF samples from patients with neuroborreliosis in comparison with the controls. MMP 9 activity was induced in vitro in a mouse macrophage cell line (RAW 264.7) when stimulated with two different genospecies of B burgdorferi (B garinii, B afzelii ). This MMP-9 activity was reduced in a dose dependent manner when macrophages stimulated with B burgdorferi were coincubated with NF-kappaB SN50, a cell permeable peptide which inhibits the translocation of NF-kappaB into the nucleus of stimulated cells. The data show that (1) MMP-9 activity is present in the CSF of patients with neuroborreliosis, (2) macrophages stimulated with B burgdorferi are a possible source of MMP-9 increase, and (3) activation of NF-kappaB may play a part in the upregulation of MMP-9 by B burgdorferi. PMID- 10675224 TI - Histological subtypes of symptomatic central nervous system tumours in Singapore. AB - The objective was to identify the different subtypes of symptomatic CNS tumours that are encountered in Singapore. Our hospital pathology and operative records from 1994 to 1998 were reviewed and information regarding all patients who underwent biopsy or resection as part of their diagnostic and therapeutic evaluation was extracted. Only histologically confirmed tumours were included in this analysis. Meningiomas made up the largest subgroup of tumours, accounting for 35.1% of all tumours. In order of decreasing frequency, the remaining most often reported histologies were pituitary adenomas (11.8%), secondary neoplasms (10%), tumours of nerve sheath (9.4%), glioblastoma multiforme (9.3%), astrocytomas including anaplastic, diffuse and pilocytic (9.2%), primary CNS lymphomas (2.9%), oligodendrogliomas (2.2%), hemangioblastomas (2. 2%), craniopharyngiomas (1.7%), and embryonal tumours (1.2%). Genetic and environmental factors may be responsible for the proportionately higher than expected percentage of meningiomas seen and further study is required to identify these factors. PMID- 10675225 TI - New variant Creutzfeldt-Jakob disease: three case reports from Leicestershire. AB - Since a report in 1996 of 10 cases of Creutzfeldt-Jakob disease (CJD) with onset in a younger than usual age, a pattern of the disease has emerged. This includes early neuropsychiatric features and sensory symptoms and neurological signs such as ataxia and involuntary movements later in the course of the disease. Three patients with varied clinical presentations and disease course seen at a single neurology unit are described. The first patient was characterised by cognitive and psychiatric symptoms together with neurological signs. The second patient presented with unusual behavioural disturbance and episodes of collapse. The third patient exhibited striking psychomotor retardation and had abnormal CSF and MRI findings. All patients succumbed in a state of akinetic mutism and myoclonus. All three patients had the methionine/methionine genotype at codon 129 of the PrP gene and in two of the three patients a tonsil biopsy was performed with positive results. These two patients also tested positive for the 14.3.3. protein in the CSF. Whereas late features of the disease seem very similar in all cases, the initial presentation was variable and underlines the uncertainty of the range of the clinical phenotype. Successful diagnosis demands a high index of clinical suspicion. PMID- 10675226 TI - Idiopathic intracranial hypertension and anticardiolipin antibodies. AB - The association of idiopathic intracranial hypertension (IIH) or pseudotumour cerebri (PTC) with anticardiolipin antibodies (aCL-Abs) has been only acknowledged recently. However, its true incidence is as yet unknown. In this retrospective study, the co-occurrence of IIH and aCL-Abs was looked for among a relatively large group of patients diagnosed with IIH or PTC in the neuro ophthalmology clinic during the years of 1992-8. All patients underwent routine blood tests and the presence of activated protein C resistance and protein S and protein C deficiency were recorded. ACL-Abs were determined in all patients. The co-occurrence of IIH and aCL-Abs was found in three out of 37 patients (8.1%), which is higher than the incidence of aCL-Abs in the general population but considerably lower than that reported in two previously published studies. The aCL-Ab positive patients in our series were significantly older and thinner than those in whom antibodies were undetected. In conclusion, it seems that patients with this association should be considered as a unique subgroup of IIH. PMID- 10675227 TI - Improvement of memory guided saccades in parkinsonian patients by high frequency subthalamic nucleus stimulation. AB - Recent studies in the monkey suggest that the subthalamic nucleus (STN) is involved in control of eye movement, yet its functional significance in humans is unknown. Saccadic eye movements were studied in eight parkinsonian patients treated by bilateral electrical stimulation of the STN. STN stimulation improved the accuracy of memory guided saccades but not of reflexive visually guided saccades and had no effect on the antisaccade task. This study shows that, by contrast with levodopa, STN stimulation improves memory guided saccade deficits, and illustrates for the first time in humans the role of the STN in the control of purposive saccades. PMID- 10675228 TI - Neurological truant. PMID- 10675229 TI - Myopathy due to primary systemic amyloidosis. PMID- 10675231 TI - Intra-operative diagnosis of CNS tumours PMID- 10675230 TI - Monocular visual loss and platelet fibrin embolism to the retina. PMID- 10675232 TI - MRI of the brain. Normal anatomy and normal variants PMID- 10675234 TI - Clinical diagnosis and management of Alzheimer's disease, 2nd edition PMID- 10675233 TI - Everything you need to know about old age psychiatry PMID- 10675235 TI - Dementia handbook PMID- 10675237 TI - Effect of laparoscopic fundoplication on gastroesophageal reflux disease-induced respiratory symptoms. AB - Laparoscopic fundoplication controls heartburn and regurgitation, but the effects on the respiratory symptoms of gastroesophageal reflux disease (GERD) are unclear. Confusion stems from difficulty preoperatively in determining whether cough or wheezing is actually caused by reflux when reflux is found on pH monitoring. To date, there is no proven way to pinpoint a cause-and-effect relationship. The goals of this study were to assess the following: (1) the value of pH monitoring in establishing a correlation between respiratory symptoms and reflux; (2) the predictive value of pH monitoring on the results of surgical treatment; and (3) the outcome of laparoscopic fundoplication on GERD-induced respiratory symptoms. Between October 1992 and October 1998, a total of 340 patients underwent laparoscopic fundoplication for GERD. From the clinical findings alone, respiratory symptoms were thought possibly to be caused by GERD in 39 patients (11%). These 39 patients had been symptomatic for an average of 134 months. They were all taking H2-blocking agents (21%) or proton pump inhibitors (79%). Seven patients (18%) were also being treated with bronchodilators, alone (3 patients) or in combination with prednisone (4 patients). Median length of postoperative follow-up was 28 months. In 23 patients (59%) a temporal correlation was found during 24-hour pH monitoring between respiratory symptoms and episodes of reflux. Postoperatively heartburn resolved in 91% of patients, regurgitation in 90% of patients, wheezing in 64% of patients, and cough in 74% of patients. Cough resolved in 19 (83%) of 23 patients in whom a correlation between cough and reflux was found during pH monitoring, but in only 8 (57%) of 14 of patients when this correlation was absent. Cough persisted postoperatively in the two patients who did not cough during the study. These data show that pH monitoring helped to establish a correlation between respiratory symptoms and reflux, and it helped to identify the patients most likely to benefit from antireflux surgery. Following laparoscopic surgery, respiratory symptoms resolved in 83% of patients when a temporal correlation between cough and reflux was found on pH monitoring; heartburn and regurgitation resolved in 90%. PMID- 10675236 TI - Telomerase reverse transcriptase expression is increased early in the Barrett's metaplasia, dysplasia, adenocarcinoma sequence. AB - Barrett's esophagus is a multistage polyclonal disease that is associated with the development of adenocarcinoma of the esophagus and esophagogastric junction. Telomerase activation is associated with cellular immortality and carcinogenesis, and increased expression of the telomerase reverse transcriptase catalytic subunit (hTERT) has been used for the early detection of malignant diseases. To identify biomarkers associated with each stage of the Barrett's process, relative mRNA expression levels of hTERT were measured using a quantitative reverse transcription-polymerase chain reaction method (ABI 7700 Sequence Detector (TaqMan system) in Barrett's intestinal metaplasia (n = 14), Barrett's dysplasia (n = 10), Barrett's adenocarcinoma (n = 14), and matching normal squamous esophagus tissues (n = 32). hTERT expression was significantly increased at all stages of Barrett's esophagus, including the intestinal metaplasia stage, compared to normal tissues from patients without cancer (intestinal metaplasia vs. normal esophagus, P <0.0001; dysplasia, P = 0.001; adenocarcinoma, P = 0.007; all Mann-Whitney U test ). hTERT expression levels were significantly higher in adenocarcinoma tissues than in intestinal metaplasia tissues (P = 0.003), and were higher in dysplasia compared with intestinal metaplasia tissues (P = 0.056). hTERT levels were also significantly higher in histologically normal squamous esophagus tissues from cancer patients than in normal esophagus tissues from patients with no cancer (P = 0.013). Very high expression levels ([hTERT x 100: beta-actin] >20) were found only in patients with cancer. These findings suggest that telomerase activation is an important early event in the development of Barrett's esophagus and esophageal adenocarcinoma, that very high telomerase levels may be a clinically useful biomarker for the detection of occult adenocarcinoma, and that a widespread cancer "field" effect is present in the esophagus of patients with Barrett's cancer. PMID- 10675238 TI - Nonsteroidal anti-inflammatory drugs attenuate proliferation of colonic carcinoma cells by blocking epidermal growth factor-induced Ca++ mobilization. AB - Numerous studies suggest that nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit colorectal carcinogenesis. We have previously reported that NSAIDs, in human colonic carcinoma cells (Caco-2), attenuate epidermal growth factor (EGF) induced cellular proliferation through a process independent of their inhibitory effects on prostaglandin synthesis. Furthermore, separate studies have also suggested that NSAIDs inhibit EGF-induced store-operated Ca++ influx. Thus we developed the hypothesis that NSAIDs may limit the activity of EGF by altering intracellular Ca++ ([Ca++]i) mobilization. Serum-deprived Caco-2 cells were employed for all experimentation. [Ca++]i was measured with Fluo-3 and extracellular Ca++ influx was monitored by quenching Fluo-3 fluorescence with Mn++. Proliferation was quantitated with two assays: cellular nucleic acid and total protein content. Caco-2 cells exposed to EGF demonstrated an initial increase in [Ca++]i which was blocked by neomycin, an inhibitor of IPsubscript 3 generation, and the phospholipase C inhibitor U73122 but not U73343 (inactive control). This was followed by sustained extracellular Ca++ influx, which was attenuated with calcium-free buffer (-Ca++), the store- operated Ca++ channel blocker lanthanum, indomethacin, ibuprofen, and aspirin. In subsequent studies, cells were treated with either serum-free media or EGF +/- the aforementioned inhibitors, and again serum starved. Cells exposed to EGF +/- the inactive phospholipase C inhibitor U73343 demonstrated a significant increase in nucleic acid and protein. However, proliferation induced by EGF was not observed when [Ca++]i elevation was prevented by blocking either internal Ca++ store release via phospholipase C/IPsubscript 3 or sustained Ca++ influx through store-operated Ca++ channels. Sustained [Ca++]i elevation, as induced by EGF, appears to be required for mitogenesis. These data support our premise that one mechanism whereby NSAIDs may attenuate colonic neoplasia is by blocking EGF-induced Ca++ mobilization. PMID- 10675239 TI - Hepatic resection using intermittent vascular inflow occlusion and low central venous pressure anesthesia improves morbidity and mortality. AB - Hepatic resection results in significant morbidity and mortality primarily related to intraoperative blood loss. Intermittent vascular inflow occlusion (VO) and low central venous pressure (CVP) during hepatectomy have been used to reduce blood loss. To determine the benefit of VO and low CVP, we reviewed the outcomes of 168 consecutive patients who underwent liver resection. The results of 78 patients who had undergone hepatic resection between 1993 and 1998 with the use of VO and low CVP (post-VO/CVP) were compared to the previous 90 patients who had undergone hepatectomy without VO and low CVP (pre-VO/CVP) between 1979 and 1992. Hepatectomies were performed for metastatic disease (65%), hepatoma (20%), and benign tumors (15%). Resections included 18 trisegmentectomies, 67 lobectomies, and 83 segmental resections. Patients in both groups were similar with regard to extent of resection. Post-VO/CVP patients had significantly lower median estimated blood loss (725 ml vs. 2300 ml, P <0.001), less postoperative morbidity (10.3% vs. 22. 2%, P = 0.038), lower in-hospital mortality (2.6% vs. 10%, P = 0. 050), fewer days in the intensive care unit (1.6 +/- 0.1 days vs. 5. 6 +/- 1.2 days, P = 0.003), and shorter overall length of stay (8.0 +/- 0.5 days vs. 15.0 +/- 1.6 days, P <0.001) than pre-VO/CVP patients. These data suggest that VO and low CVP during liver resection may improve patient outcomes. PMID- 10675240 TI - Isolated right segmental hepatic duct injury: a diagnostic and therapeutic challenge. AB - Biliary leaks and injuries are not an uncommon occurrence following laparoscopic cholecystectomy. Bile leaks associated with the biliary anatomic variant of a low inserting right segmental hepatic duct can be particularly difficult to diagnose in that results of endoscopic retrograde cholangiography (ERC) are usually interpreted as "normal" with no leaks demonstrated. The aim of this study was to describe a single institution's experience with nine patients with biliary leaks associated with this anatomic variant and to discuss their management. A retrospective analysis of the hospital records of all patients with bile duct injuries managed at a single institution between 1980 and July 1998, inclusive, was performed. Nine patients were identified as having an isolated right segmental hepatic duct injury associated with a biliary leak. Seven (78%) of the nine patients had undergone a laparoscopic cholecystectomy, whereas the remaining two patients (22%) had undergone an open cholecystectomy. All of the patients had undergone endoscopic retrograde cholangiography at outside institutions, the results of which had been interpreted as normal with no apparent leaks. The median interval from the time of cholecystectomy to referral was 1.4 months. All patients were managed with initial percutaneous access of the involved right segmental biliary system, with placement of a percutaneous transhepatic stent. After the biliary leak was controlled, all patients underwent Roux-en-Y hepaticojejunostomy to the isolated biliary segment. All patients had an uncomplicated postoperative course. There were no postoperative anastomotic leaks. Postoperative stenting was maintained for a mean of 8 months. Six (67%) of the nine patients had a long-term successful outcome with minimal or no symptoms. In three patients, recurrent symptoms with pain and/or cholangitis developed at a mean of 34 months. All three patients underwent percutaneous cholangiography, which demonstrated an anastomotic stricture, and all were managed with percutaneous balloon dilatation with a successful outcome. Currently eight (89%) of the nine patients are asymptomatic, with a mean follow-up of 70.4 months (range 12 to 226 months). One patient had intermittent right upper quadrant pain with normal liver function tests but has not required intervention. Isolated right segmental hepatic ductal injury with biliary leakage is an uncommon complication following laparoscopic cholecystectomy. A diagnostic dilemma is created by the presence of a bile leak with a normal endoscopic retrograde cholangiogram. Management begins with percutaneous access of the transected isolated ductal system followed by reconstruction as a Roux-en-Y hepaticojejunostomy. PMID- 10675241 TI - Anatomic segmental hepatic resection is superior to wedge resection as an oncologic operation for colorectal liver metastases. AB - Hepatic wedge resection for colorectal liver metastasis has been reported to have a high incidence of positive surgical margins. Anatomic segmental resection is now widely practiced, although there are few data comparing segmental and wedge resection in terms of tumor clearance or long-term outcome. There were 267 patients who underwent liver resection for metastatic colorectal cancer between July 1985 and October 1998 at our institution who had either a wedge (n = 119) or segmental (n = 148) resection. Patient, tumor, and treatment data were compared, actuarial survival was determined, and prognostic factors were analyzed. Anatomic segmental resection was associated with similar blood loss, operative time, and complications as wedge resection. Segmental resection had a significantly lower rate of positive margins (2% vs. 16%) compared to wedge hepatectomy (P <0.001). On univariate analysis, segmentectomy resulted in longer survival with a median of 53 months vs. 38 months for wedge hepatectomy (P = 0.015). Preoperative carcinoembryonic antigen level, positive margin of resection, and the presence of extrahepatic disease independently predicted survival on multivariate analysis. Anatomic segmental resection is a safe procedure and is superior to wedge resection as an oncologic operation for colorectal liver metastasis because it results in better tumor clearance and improved survival. PMID- 10675242 TI - Inflammatory cytokines alter human gallbladder epithelial cell absorption/secretion. AB - Gallbladder inflammation is an early feature of gallstone formation in animal models. The inflammatory response is associated with increases in myeloperoxidase and interleukin (IL)-1 activities in the gallbladder wall. The present studies were designed to determine whether inflammatory cytokines directly affect gallbladder epithelial cell absorptive function. Studies were performed using cultured human gallbladder epithelial cells derived from a well-differentiated gallbladder carcinoma. Confluent monolayers were exposed to interleukin-1 (IL 1alpha), IL-1alpha plus its specific receptor inhibitor IL-1ra, tumor necrosis factor (TNF-alpha), lipopolysaccharide, or prostaglandin E2. Unidirectional sodium and chloride fluxes were measured and used to calculate net ion fluxes. Compared to control monolayers, lipopolysaccharide, prostaglandin E2, IL-1alpha, and TNF-alpha decreased mucosal-to-serosal and net sodium and chloride fluxes and increased serosal-to-mucosal movement of sodium and unmeasured ions. The effects of IL-1alpha were completely inhibited by its specific receptor antagonist IL 1ra. Similar to the proinflammatory agents lipopolysaccharide and prostaglandin E2, the inflammatory cytokines IL-1alpha and TNF-alpha directly affected gallbladder epithelial cell absorptive function. Because normal gallbladder absorptive function is protective against gallstone formation, alterations in absorptive function due to inflammation in the gallbladder wall may play a role in gallstone pathogenesis. PMID- 10675243 TI - Role of cytosolic phospholipase A2 in cytokine-stimulated prostaglandin release by human gallbladder cells. AB - Eicosanoids are involved in gallbladder inflammation, epithelial water transport, and mucous secretion. Phospholipase Asubscript2 enzymes liberate arachidonic acid from membrane phospholipids for the synthesis of eicosanoids. The purpose of this study was to determine the effect of selective cytoplasmic and secretory phospholipase A2 inhibitors on basal and stimulated arachidonic acid and prostaglandin E2 release in gallbladder cells. Western immunoblotting was employed to evaluate both cytosolic and secretory phospholipase A2 enzymes in human gallbladder cells. Cells were incubated for 22 hours with (3)H-labeled arachidonic acid. Arachidonic acid and prostaglandin E2 release was then measured in the supernate after 2 hours of exposure to human interleukin-1beta, alone or after pretreatment for 1 hour with the inhibitors. Unstimulated gallbladder cells express both 85 kDa cytosolic and 14 kDa secretory phospholipase A2++. The 85 kDa phospholipase A2 was induced by interleukin-1beta, whereas there was no apparent change in secretory phospholipase A2 enzyme concentrations. Both the secretory phospholipase A2 inhibitor p-bromophenylacyl bromide and the cytosolic phospholipase A2 inhibitor arachidonyl trifluoromethyl ketone decreased basal and interleukin-1beta-stimulated arachidonic acid release. In contrast, only inhibition of cytosolic phospholipase A2 led to a decrease in interleukin-1beta stimulated prostaglandin E2 release. Basal and interleukin-1beta-stimulated arachidonic acid release appears to be the result of the activity of both cytosolic and secretory phospholipase A2. Interleukin-1beta-stimulated prostaglandin E2 release appears to be dependent on the activity of cytosolic phospholipase A2. PMID- 10675244 TI - Predicting the need for colectomy in pediatric patients with ulcerative colitis. AB - Total colectomy is curative for ulcerative colitis. However, many pediatric patients are medically managed and may not require surgery. There are currently no available criteria to identify children who will benefit from early colectomy. The purpose of this review was to identify criteria associated with the need for colectomy. A 15-year review of patients at a major pediatric center with biopsy proved ulcerative colitis was conducted. Age at the time of the first symptom, diagnosis, and surgery were recorded as well as steroid dependence, site of disease, extraintestinal manifestations, and family history. Seventy-three patients ranging in age from 1 to 18 years were identified. Thirty-seven patients (50.1%) required total colectomy before the age of 18. The average patient age at the time of the first documented symptom was 11.3 +/- 0.5 years. Among patients who were steroid dependent and had pancolitis, 73% required colectomy. Patients with these factors failed medical management 77% (27 of 35) of the time, and colectomy was performed within 3 years of diagnosis. The combination of steroid dependence and pancolitis was associated with an increased need for colectomy. In pediatric patients with these factors, early colectomy may limit the need to endure prolonged courses of medications and the disability allied with this disease. PMID- 10675245 TI - The jejunal pouch as a rectal substitute after proctocolectomy. AB - Our hypothesis was that a jejunal pouch used as a rectal substitute after proctocolectomy would slow enteric transit, delay defecation, and decrease stool frequency compared to an ileal pouch so used. Twelve dogs underwent proctocolectomy; six had a jejunal pouch-distal rectal anastomosis and six had an ileal pouch-distal rectal anastomosis. After recovery, postprandial mouth-to-anus transit was slower in jejunal pouch dogs (253 +/- 18 minutes [mean +/- SEM]) than in ileal pouch dogs (112 +/- 7.9 minutes; P <0.05). Moreover, jejunal pouch dogs passed only 4.1 +/- 0.3 stools during the 12 hours after eating, whereas ileal pouch dogs passed 6.3 +/- 0. 9 stools (P <0.05). The mean frequency of proximal ileal pacesetter potentials after feeding was less in jejunal pouch dogs (12 +/- 0.4 cycles/min) than in ileal pouch dogs (16 +/- 0.3 counts/min; P = 0. 01), and jejunal pouches had more action potentials (jejunal = 82% +/- 4.3% of pacesetter potentials had action potentials, ileal = 61% +/- 3.0%; P <0.05). In contrast, gastric emptying and pouch motility, emptying, mucosal integrity, and bacteriologic and histologic properties were similar in the two groups of dogs. We concluded that the jejunal pouch operation slowed enteric transit, delayed defecation, and decreased postprandial stooling compared to the ileal pouch operation. PMID- 10675247 TI - Economic models of animal communication. AB - Many models of animal signal evolution fail to incorporate an explicit strategy for receivers prior to the evolution of signals. When reasonable assumptions are made for such strategies, we have shown that there is a minimal accuracy of signal coding that is required before receivers should attend to signals (Bradbury & Vehrencamp 1998, Principles of Animal Communication). Depending upon the relative payoffs of correct and incorrect decisions by receivers, this minimal accuracy can be quite high. Here we use this result to explain why so many signals appear to be traits that provided useful information to receivers before becoming ritualized into signals. Our model also supports one prediction of sensory drive models: that latent preferences may selectively favour some signal precursors over others. However, it imposes a serious constraint on sensory drive by requiring that there be sufficient benefits to a receiver to compensate for the costs of disrupting the optimal receiver strategy used before exploitation. Finally, we discuss the overlap between signal honesty and accuracy and show how senders that completely disagree with receivers about appropriate receiver decisions may still benefit by providing moderately honest and accurate signals. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675246 TI - Prospective randomized trial of early initiation and hospital discharge on a liquid diet following elective intestinal surgery. AB - Length of hospital stay after elective intestinal surgery may be related to patient tolerance of a diet. We hypothesized that early initiation and discharge home on a clear liquid diet would decrease the length of hospital stay without increasing morbidity. The aim of this study was to determine if early initiation and discharge on a clear liquid diet decreases the length of hospital stay and is safe. Forty-four patients were randomly assigned to either a standard diet or a clear liquid diet. A standard diet (n = 17) was begun after the passage of flatus or stool, and consisted of clear liquids to a volume of approximately 750 ml, then three solid meals, and discharge thereafter. Patients randomized to a clear liquid diet (n = 27) received 30 ml/hr of clear liquids on postoperative day 2, unlimited clear liquids on postoperative day 3, and were dismissed on a clear liquid diet on postoperative day 4. All patients were followed by a daily telephone call and clinic visit. The primary outcome variable was length of hospital stay. The incidence of postoperative intestinal-related sequelae, complications, and readmission rates did not differ between groups. Postdischarge intestinal symptoms were common in both groups but tended to resolve faster in the patients on a standard diet. The length of hospital stay was decreased in the patients on a clear liquid diet compared to those on a standard diet (6.1 +/- 1.1 days vs. 4.4 +/- 0.2 days; P = 0.09), but total hospital costs did not differ. Early initiation and hospital discharge on a clear liquid diet after elective intestinal surgery decreases the length of hospital stay and is safe. PMID- 10675248 TI - Assessing the degree of association in groups of animals using the concept of entropy. AB - Nearest-neighbour analysis is commonly used to calculate indices of aggregation in groups of animals. It has several problems, however, including lack of data independence and, when studying groups of animals penned at high densities, the difficulty of determining a given individual's nearest neighbour. We describe an entropy-based method to assess the degree of association (or segregation) of groups of animals. We show that this method gives more information and is more sensitive than the nearest-neighbour technique. An example with a particular experimental situation (mixing groups of lambs) is presented. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675249 TI - Females of the lekking great snipe do not prefer males with whiter tails. AB - A previous experimental study of great snipe, Gallinago media, has reported an effect on male mating success of the amount of white in their tails. That result is one of a very limited set of existing experimental results supporting a female mate preference for a morphological trait in animals. However, a later observational study did not find any correlation between amount of white and male mating success. If females sample a limited number of males, their preferences need not result in strong relationships between mating success and trait values in males, possibly explaining the failure to find the predicted correlation. Yet, females of lekking species are thought to have ample opportunities for mate sampling. To resolve these contrasting results, we present in this paper (1) a larger correlational study (several leks during 10 years) showing no relationship between male mating success and whiteness of tails (measured in several ways), and most importantly (2) evidence that individual females do not mate predominantly with males with very white tails among those males that each female samples. These results show that females do not prefer males with whiter tails as mates, within the contemporary natural variation in the trait. They also show that there is no sexual selection of the trait at present. This does not necessarily imply that white tails are not a sexually selected adaptation in males, but the mechanisms are likely to have been different from direct mate choice of whiter tails per se. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675250 TI - Did aggregation favour the initial evolution of warning coloration? A novel world revisited. AB - From experiments using novel prey signals to avoid innate reactions to traditional signals, Alatalo & Mappes (1996, Nature, 382, 708-710) concluded that gregariousness would have selected for warning coloration as it originated for the first time, whereas a solitary prey distribution would not. We have investigated this suggestion in experiments using the same novel prey and background symbols and wild-caught great tit, Parus major, predators. We compared the attack rate on cryptic unpalatable and aposematic unpalatable prey in either a solitary or an aggregated treatment. In the aggregated treatment we found no difference in attack rate on cryptic and aposematic prey. In the solitary treatment the attack rate on aposematic prey was significantly lower after one attack and at the end of the experiment. Thus, we conclude that, in so far as these experiments mimic an original predator-prey relationship, they do not give support to the idea that aggregation would have favoured the evolution of warning coloration in unpalatable prey. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675251 TI - Adoption of chicks and the level of relatedness in common gull, Larus canus, colonies: DNA fingerprinting analyses. AB - In common gull colonies on islands of the Vistula River, Poland, adoption of chicks is common. In 1997, we observed 81 chicks from 35 nests. Of these, 19 (23.4%) left their natal broods and were adopted by other pairs. Another 11 (31.4%) were driven from the foreign territory by the owners. Foreign chicks were adopted by 15 pairs (42.9%). Eleven pairs (31.4%) drove foreign chicks from the territory. To test if the frequent adoptions in these colonies could be explained by kin selection or the occurrence of kin groups, we calculated band-sharing coefficients and genetic relatedness (r) between interacting birds (neighbours and non-neighbours). Adults that adopted were most often neighbours of the biological parents of adopted chicks, whereas spatially segregated birds, nesting further away, usually drove off the chicks. Band-sharing coefficients between males, but not females, were higher with decreasing internest distances. The band sharing coefficients for adopted chicks and foster parents were significantly higher than for adopted chicks and randomly selected, spatially segregated pairs from the same and another colony. Band-sharing coefficients of adopted chicks and adopting neighbours (males: r=0.20; females: r=0.16) also tended to be higher than those of rejected chicks and rejecting neighbours (both sexes: r=0.08). Our results suggest that kin groups of neighbours do occur in common gull colonies. Such social structure might lead to indirect inclusive fitness benefits of adopting pairs. Differences in genetic similarity between chicks and adopting or rejecting neighbours show that at least in common gulls we should consider kin altruism as a factor in adoptions. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675252 TI - Green plants in starling nests: effects on nestlings. AB - European starlings, Sturnus vulgaris, intermingle fresh herbs, especially species rich in volatile compounds, with their otherwise dry nest material. In this field study we investigated whether these herbs reduce ectoparasites and thereby protect nestlings (the nest protection hypothesis). We also considered whether volatile compounds in herbs improve the condition of nestlings (the drug hypothesis). As measures of condition we used body mass, haematocrit levels and immunological parameters. We replaced 148 natural starling nests with artificial ones: half contained herbs and half (controls) contained grass. The ectoparasite loads (mites, lice, fleas) in herb and control nests were indistinguishable. However, nestlings in herb nests weighed more and had higher haematocrit levels at fledging than nestlings in control nests. Fledging success was similar in herb and control nests, but more yearlings from herb nests were identified in the colony the year after hatching. The response of the immune system when challenged with phytohaemagglutinin did not differ in nestlings from herb and control nests. Nestlings from herb nests had more basophils and fewer lymphocytes in their blood than those from control nests, while the eosinophil and heterophil counts did not differ. We conclude that herbs do not reduce the number of ectoparasites, but they improve the condition of nestlings, perhaps by stimulating elements of the immune system that help them to cope better with the harmful activities of ectoparasites. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675253 TI - Female-mediated differential sperm storage in a fly with complex spermathecae, Scatophaga stercoraria. AB - Multiple spermathecae potentially allow selective sperm use, provided that sperm from rival males are stored differentially, that is, in different proportions across storage compartments. In the yellow dung fly, Scatophaga stercoraria, females have three spermathecae arranged as a doublet and singlet. To test whether females store the sperm of rival males actively and differentially, we mated fixed male pairs to three females. After copulation, females were (1) dissected immediately before they could start laying a clutch of eggs, (2) left awake for 30 min but prevented from oviposition, or (3) anaesthetized with carbon dioxide for 30 min to interfere with the muscular control presumably required for sperm transport from the site of insemination to the spermathecae. For each female, we estimated the proportion of the second male's sperm stored in her spermathecae (S(2)value), using sperm length as a male marker. After copulation, the S(2)values in the singlet and doublet spermathecae differed significantly, indicating differential sperm storage during copulation. Postcopulatory treatment affected differential sperm storage significantly. Females dissected immediately had lower S(2)values in the doublet than in the singlet spermatheca, while females left awake showed the reverse pattern for the same two males. This reversal did not occur when females were treated with carbon dioxide. The results indicate differential storage of sperm from different males during copulation and that female muscular activity can affect storage and separation of competing ejaculates beyond copulation. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675254 TI - Sperm trading in a hermaphroditic flatworm: reluctant fathers and sexy mothers. AB - When matings are frequent and received sperm are digested, hermaphrodites should trade sperm when mating. We investigated sperm trading in the flatworm Schmidtea (Dugesia) polychroa and manipulated mating interests to investigate its possible causes. In 106 mating pairs consisting of nonisolated individuals, no sperm donation in either direction (35%) and reciprocal exchange (38%) were more common than expected by chance, whereas unilateral transfer (27%) was less frequent, confirming sperm trading. The amount of sperm donated depended on the availability of self-sperm, not on the amount received. Animals with more allosperm from previous matings had more self-sperm and consequently donated more. This suggests that sperm digestion boosts sperm production. In a second experiment, 'mixed-interest' pairs consisting of a nonisolated (N) and an isolated individual (I), NxI, were compared with IxI and NxN pairs. Whereas IxI pairs were eager and NxN reluctant to mate, NxI pairs showed an intermediate likelihood of mating. Whereas NxN pairs traded sperm, the other two groups did not. The change in behaviour in N individuals in the NxI treatment suggests precopulatory assessment and mating in relation to phenotypic mate quality. Isolated individuals are attractive, presumably because they donate large sperm clumps unconditionally and contain fewer allosperm, implying reduced sperm competition. The reduced reluctance in N individuals to mate with, and to inseminate, previously isolated partners suggests that female quality is an important factor in male sperm donation decisions. Hence, S. polychroa may be choosier than previously assumed. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675255 TI - Finescale topographical correlates of behavioural investment in offspring by female grey seals, Halichoerus grypus. AB - Grey seals breed colonially on substrates ranging from ice to rocky or sandy beaches. Clear differences in seal behaviour patterns exist among such broad classes of breeding habitat. However, finer scale topographical variation is likely to influence individual behaviour with consequences for pupping success. We examined topographical influences on the behaviour of breeding female grey seals by quantifying topography at a subseal size resolution. Using submetre resolution digital terrain models of two sites within a rocky breeding colony, we compared site topography in relation to observed differences in female behaviour at these sites. Females at both sites preferred breeding close to water (standing pools or sea) and frequently commuted between their pups and water. Topographical models indicated that one site was more costly for seals in terms of their locations and movements within the site. This was due to a lack of low-elevation land adjacent to the main access points from the sea and the reduced availability of pools. Females at this site showed reduced pup attendance and an increase in energetically costly behaviours, whilst females at the lower-cost site spent more time interacting with their pups and resting. These topographically induced behavioural differences are likely to affect the quantity and quality of pup provisioning by mothers and influence individual pupping site selection. Less costly sites are likely to be colonized preferentially and by larger, older and more dominant females, potentially generating finescale spatial heterogeneity in female quality within the breeding colony. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675256 TI - Components of lifetime mating success and body size in males of a scrambling damselfly. AB - Sexual selection is hypothesized to favour small body size in males of scrambling species, that is, those in which males obtain matings by actively searching for females. I tested this hypothesis in a natural population of the scrambling emerald damselfly, Lestes sponsa. Mating efficiency (matings/visit to the pond) was the most important factor explaining variation in male lifetime mating success (LMS; 71%). This suggests a large potential for sexual selection. Path analysis of male LMS suggested a quality factor that positively affected both mating efficiency and life span. In contrast with the small-male mating advantage hypothesis, part of this potential for sexual selection was realized as stabilizing selection on male body size, indicating that there may also be a lower limit to body size for mating efficiency. This also illustrates that the constancy of body size may be explained by sexual selection alone. Survival explained about 20% of the variation in LMS and random processes were potentially important for determining LMS. My results show the problems of using mating efficiency as a measure for the intensity of sexual selection and the need to distinguish between potential and realized selection pressures, especially when comparing the importance of natural and sexual selection. I discuss mechanisms that may have caused the intermediate-male mating advantage in this scrambling species. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675257 TI - The transvestite serpent: why do male garter snakes court (some) other males? AB - In large mating aggregations of red-sided garter snakes, Thamnophis sirtalis parietalis, in Manitoba, male courtship is directed not only to females, but also to other males with female-like skin lipids ('she-males'). We show that 'she maleness' is an intrinsic property of a male rather than an artefact of lipid transfer from females, and that male-male courtship is very common in the field. She-males were distinctive in terms of appearance (they were heavier than other males and more often covered with mud), behaviour (they were inactive and rarely courted females) and performance (they were slow crawlers, ineffective courters and easily outcompeted by other males in mating trials). 'She-maleness' was not a characteristic of a particular subset of males, as envisaged in previous work; instead, it was a transitory phase that most (perhaps all) male snakes passed through soon after they first emerged from the winter den. Recently emerged males spent their first day or two relatively inactive, while restoring physiological functions (including locomotor performance and courtship ability). Experimental application of female skin lipids on to males dramatically decreased courtship levels of the recipient snakes. Thus, recently emerged males may derive two kinds of benefit from mimicking female skin lipids. First, female mimicry 'switches off' the male's own (energetically expensive) courtship at a time when that courtship would be unproductive. Second, it may disadvantage his rivals by distracting them from females, and increasing their energy expenditure. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675258 TI - Acoustic satellite behaviour in the Australian bushcricket Elephantodeta nobilis (Phaneropterinae, Tettigoniidae, Orthoptera). AB - Male and female Elephantodeta nobilis duet with the female responding to the male's long and complex call. The duetting male's call consisted of four parts, described here as parts A, B, C and D. We found that the female replied 570 ms after the male's D pulse, which followed the extended part B and short click of part C. Noncalling males were attracted to the duet and often used satellite tactics by inserting a volley of clicks 200 ms before the alpha male's D pulse. Satellite males used part C of the alpha male song to cue their own call and this inserted call induced females to reply earlier compared with the alpha male call alone. Alpha males often extended their calls with additional D-type calls and so we examined the effectiveness of these calls as countermeasures to satellite calling. There was no influence of this alpha strategy on the satellite's propensity to call although more calls from the alpha male did cause the female to reply more frequently. We also examined the effect of relative intensity of alpha and satellite calls on the female's reply. Reduced satellite intensity increased the variance in the timing of the female response. Finally, we tested the effectiveness of the satellite's call on female phonotaxis within a two speaker arena. Although females preferred the alpha male they were nevertheless attracted to the satellite calls regardless of the latter's relative intensity. We discuss the possible role of satellite calling as a novel conditional strategy. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675259 TI - Free female mate choice in house mice affects reproductive success and offspring viability and performance. AB - We tested a critical assumption of sexual dialectics theory (Gowaty 1997, Feminism and Evolutionary Biology, Chapman & Hall) using house mice, Mus musculus. We asked if female house mice accrue viability benefits for their offspring when they mate with males they prefer versus with males they do not prefer. Our experiment was designed to eliminate or control other mechanisms of reproductive competition besides female mate choice. After allowing females to discriminate behaviourally between two males, which were at random with respect to phenotypic variation discriminating females were paired with preferred (P) or nonpreferred (NP) males. We then tested whether females mating with males they preferred had offspring of higher viability than females mating with nonpreferred males. In pairwise comparisons, we tested for differences in offspring performance in dominance contests and in nest-building skill. At weaning, we exposed half of the pups to cold stress. We tested progeny performance and viability in the laboratory or in outdoor field enclosures. In comparison to P females, NP females produced significantly fewer litters. Sons from P matings were socially dominant to sons from NP matings. Adult offspring from P matings built better nests than those from NP matings. In field enclosures significantly fewer NP than P offspring survived to 60 days after introduction. Male and female progeny from P matings established larger home ranges and constructed better nests than progeny from NP matings. This is the first demonstration of progeny viability differences for females allowed to express mate preferences between males presented to them at random. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675260 TI - Egg-dumping lace bugs preferentially oviposit with kin. AB - Egg dumping, or abandoning eggs and young to the care of other conspecifics, results in an extreme form of alloparental care. It is unclear, however, if egg dumpers discriminate among kin and nonkin egg recipients. In the lace bug Gargaphia solani (Heteroptera: Tingidae), some females with eggs (guards) also accept and defend eggs of conspecifics. Other females (egg dumpers) abandon their offspring after oviposition, leaving a single guard as the caregiver. We asked if egg dumpers preferentially dump their eggs among unguarded eggs of kin or nonkin. When given a choice between dumping among eggs of full siblings and eggs of nonsiblings, most eggs (67%) were dumped with full siblings' eggs. Furthermore, egg dumpers were just as likely to oviposit among eggs of kin with whom they had interacted on a shared host plant during juvenile development as they were to oviposit with kin reared on different host plants. Thus, egg dumpers discriminate kin by using cues associated with eggs, and such cues are not likely to be acquired through interaction on a common host plant environment. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675261 TI - Territory acquisition in loons: the importance of take-over. AB - We examined patterns of territory acquisition and reconnaissance in common loons, Gavia immer, from northern Wisconsin. Among all territory acquisitions, 41.5% occurred through passive occupation of territories left vacant after the death or desertion of a previous resident, 17% constituted founding of new territories and the remaining 41.5% came about through take-over: either usurpation of defended territories or appropriation of territories before the seasonal return of previous owners. Take-overs occurred in both sexes, but individuals acted alone, never in pairs. Displaced breeders usually took refuge on undefended lakes near their former territories; about half of these loons later regained former territories through passive occupation or took possession of new territories elsewhere. As predicted by the reconnaissance hypothesis, usurpations occurred most often in territories that had produced chicks during the previous 12 months, suggesting that loons use the presence or absence of chicks as a cue for territorial usurpation. Large individuals of both sexes held onto territories longer than small individuals, an indication that body size might be correlated with fighting ability. In terms of life history, loons appear to locate good territories through reconnaissance, usurp them in a subsequent year and recover from displacements by reclaiming their original territories or new ones. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675262 TI - Sperm storage by females of the polyandrous noctuid moth Heliothis virescens. AB - Female tobacco budworm moths, Heliothis virescens, generally mate with more than one male, receiving from each mate both fertilizing sperm (eupyrene) and nonfertilizing anucleate sperm (apyrene), which is thought to play a role in sperm competition. One male typically gains sperm precedence, but it is not consistently the last or the first male to mate. I investigated the mechanism of this variable pattern of paternity by examining the patterns of storage of both types of sperm in the female's spermatheca as a function of multiple mating and male phenotype. The number of stored apyrene sperm varied with mating history, being greatest in twice-mated females and least in females mated to one nonvirgin male. In contrast, only one ejaculate's worth of eupyrene sperm was stored regardless of female mating history (once or twice mated). Thus, while they store two complements of apyrene sperm, twice-mated females apparently store only one ejaculate's worth of eupyrene sperm. This biased pattern of sperm storage may contribute to the variable pattern of paternity observed in this species. Eupyrene sperm storage also correlated positively with female size, male age and spermatophore size. Finally, a new sperm storage site was identified and described. It is a bulged region in the seminal duct. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675263 TI - Can a minority of informed leaders determine the foraging movements of a fish shoal? AB - There is no information on whether the daily foraging movements of fish shoals are the result of chance, the collective will of all shoalmates, or the leadership of a few individuals. This study tested the latter possibility. Shoals of 12 golden shiners, Notemigonus crysoleucas, were trained to expect food around midday in one of the brightly lit corners of their tank. They displayed daily food-anticipatory activity by leaving the shady area of their tank and spending more and more time in the food corner up to the normal time of feeding. Past this normal time they remained in the shade, even on test days when no food was delivered. Most of these experienced individuals were then replaced by naive ones. The resulting ratio of experienced:naive fish could be 5:7, 3:9 or 1:11. On their own, naive individuals would normally spend the whole day in the shade, but in all tests the experienced individual(s) were able to entrain these more numerous naive fish out of the shade and into the brightly lit food corner at the right time of day. Entrainment was stronger in the 5:7 than in the 1:11 experiment. The test shoals never split up and were always led by the same fish, presumably the experienced individuals. These results indicate that in a strongly gregarious species, such as the golden shiner, a minority of informed individuals can lead a shoal to food, either through social facilitation of foraging movements or by eliciting following behaviour. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675264 TI - Courtship role reversal and deceptive signals in the long-tailed dance fly, Rhamphomyia longicauda. AB - We examined the function of secondary sexual characters in the role-reversed, lekking behaviour of female long-tailed dance flies, Rhamphomyia longicauda Loew (Empididae), to test the hypothesis that the degree of abdominal distention is an honest female signal about the state of egg development. Female Rhamphomyia cannot hunt for prey and they receive all of their protein from males by exchanging copulations for nuptial prey gifts. Females compete for male gifts within leks that are organized for a brief period each evening before dark. Before hovering within leks, females swallow air, inflating expandable pouches on the pleural margins of the abdomen. The result is a large saucer-like abdomen which is further exaggerated by wrapping scaled pro-, meso- and metathoracic legs along its pleural margins. Male preference for an enlarged abdomen was confirmed by suspending plastic models of varying size from monofilament lines and recording which models attracted the most males. There was a positive relationship between egg development and abdominal distention in a related species, R. sociabilis (Williston), which lacks inflatable abdominal pouches. Multiple regression showed that in R. longicauda, abdominal inflation completely masks the state of egg development. We conclude that female R. longicauda deceive mate-seeking males with the unreliable message that eggs are nearing maturation in order to obtain a protein meal in exchange for copulation. Males that fail to identify a female bearing mature eggs risk near-certain cuckoldry and an increased probability that the female will die before oviposition. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675265 TI - Cheap talk when interests conflict. AB - Most evolutionary analyses of animal communication suggest that low-cost signals can evolve only when both the signaller and the recipient rank outcomes in the same order. When there is a conflict of interest between sender and receiver, honest signals must be costly. However, recent work suggests that low-cost signals can be evolutionarily stable, even when the sender and the receiver rank outcomes in different orders, as long as the interest in achieving coordination is sufficiently great. In this paper, we extend this body of work by analysing a game theory model that shows that low-cost signals can evolve when there are conflicts of interest and no interest in coordination, as long as individuals interact repeatedly. We also present an empirical example indicating that female rhesus macaques, Macaca mulatta, use honest, low-cost, vocal signals to facilitate interactions when conflicts of interest exist. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675266 TI - Mechanisms of dispersed central-place foraging in polydomous colonies of the Argentine ant. AB - Many species of ants occupy multiple nests, a condition known as polydomy. Because of their decentralized structure, polydomous colonies may be removed from some of the constraints associated with classic central-place foraging. We used laboratory and field experiments to assess the mechanisms involved in dispersed central-place foraging in polydomous colonies of the Argentine ant Linepithema humile, a widespread invasive species. Both in the laboratory and in the field, Argentine ants established new nests at sites located near food. Laboratory colonies of L. humile redistributed workers, brood and resources among nests in response to the spatial heterogeneity of food resources. In addition, laboratory colonies formed recruitment trails between nests in the context of foraging, providing a mechanism for the transport of material between nests. This highly flexible system of allocating nests, workers and brood throughout a colony's foraging area potentially increases foraging efficiency and competitive ability. The importance of polydomy as a determinant of competitive ability is underscored by its prevalence among ecologically dominant ants, including most, if not all, highly invasive species. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675267 TI - Female European starling preference and choice for variation in conspecific male song. AB - Data from several field studies support the hypothesis that female European starlings, Sturnus vulgaris, attend to variation among the songs of conspecific males when making mate-choice decisions. However, for a variety of methodological reasons, direct evidence for female preferences based on song in starlings has been lacking. This study presents a novel technique for assaying directly female preference and choice in European starlings by using the presentation of conspecific male song as an operant reinforcer in a controlled environment. Using an apparatus in which the playback of songs from different nestboxes is under the operant control of the subject, we demonstrate how the reinforcing properties of conspecific song can be used to measure female preference and choice. The results of the study suggest three conclusions. First, female starlings prefer naturally ordered conspecific male songs over reversed songs. Second, female starlings display robust preferences for longer compared with shorter male song bouts. Behaviour in the operant apparatus varied directly with male song bout length. Third, preferences based on song bout length are sex specific. Male starlings failed to respond differentially to the same stimuli for which females showed strong preferences. These results suggest that male-male variation in song bout length is important for mate choice among starlings. In addition, we detail the use of a novel behavioural assay for measuring female preferences that can be applied to similar behaviours in other species of songbirds. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675268 TI - Male horn dimorphism in the scarab beetle, Onthophagus taurus: do alternative reproductive tactics favour alternative phenotypes? AB - In a variety of organisms morphological variation is discrete rather than continuous. Discrete variation within a sex has attracted particular interest as it is thought to reflect the existence of alternative adaptations to a heterogeneous selection environment. The beetle Onthophagus taurus shows a dimorphism for male horns: males that exceed a critical body size develop a pair of long, curved horns on their heads, while smaller males remain hornless. In this study we report on the alternative reproductive tactics used by males with these two morphologies, and present experimental and behavioural data suggesting that these alternative tactics selectively favour discretely different male phenotypes. Horned males aggressively defended tunnel entrances containing breeding females. Fights involved the use of horns, and males with longer horns were more likely to win fights. In contrast, hornless males employed nonaggressive sneaking behaviours when faced with competitively superior males. Sneaking behaviours appeared to require high degrees of manoeuvrability inside tunnels to access and mate with females despite the presence of a guarding male. Comparisons of running performances of males with identical body sizes but different horn lengths suggest that the possession of horns reduces male agility inside tunnels. Thus, horn possession confers a clear advantage to males using fighting behaviours to access females, whereas hornlessness may be favoured in males that rely primarily on sneaking behaviours. Combined, the two alternative reproductive tactics used by male O. taurus appear to favour opposite horn phenotypes, which may explain the paucity of intermediate morphologies in natural populations of O. taurus. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10675269 TI - Ligation of ICAM-1 molecules inhibits target cell-induced granule exocytosis of IL-12-activated natural killer cells. AB - The importance of cell adhesion molecules such as ICAM-1 is emphasized in cell-to cell interactions that are critical in the generation of effective immune reactions. In this study, the involvement of ICAM-1 in natural killer (NK) cell activities was characterized in IL-12-activated human NK cells. To address the question of whether ligation of ICAM-1 molecules can modulate NK cell cytolytic activities, a 4-h (51)Cr-release assay was performed after pretreatment of NK cells with R6.5 mAb (anti-human ICAM-1 mAb). Ligation of membrane ICAM-1 molecules significantly inhibited IL-12-enhanced NK cytotoxicity against K562, and the pretreatment of neutralizing soluble ICAM-1 with R6.5 mAb blocked this inhibitory effect. The involvement of Ca(2+)-dependent granular exocytosis was evaluated. BLT esterase assay demonstrated that the ligation of ICAM-1 molecules inhibited granular exocytosis of NK cells. Additionally, the ICAM-1-mediated inhibition of Ca(2+) flux in NK cells was detected using Fluo-3AM, while the pretreatment of NK cells with R6.5 mAb did not affect conjugate formation between NK and K562 cells. Collectively, these results suggest that the signals transduced from ICAM-1 molecules might be sufficient to induce inhibitory effects on NK cells. PMID- 10675270 TI - Regulation of human natural killer cell migration and proliferation by the exodus subfamily of CC chemokines. AB - Natural killer (NK) cells play an important role in innate and adaptive immune responses to obligate intracellular pathogens. Nevertheless, the regulation of NK cell trafficking and migration to inflammatory sites is poorly understood. Exodus 1/MIP-3alpha/LARC, Exodus-2/6Ckine/SLC, and Exodus-3/MIP-3beta/ELC/CKbeta-11 are CC chemokines that share a unique aspartate-cysteine-cysteine-leucine motif near their amino terminus and preferentially stimulate the migration of T lymphocytes. The effects of Exodus chemokines on human NK cells were examined. Exodus-1, -2, and -3 did not induce detectable chemotaxis of resting peripheral blood NK cells. In contrast, Exodus-2 and -3 stimulated migration of polyclonal activated peripheral blood NK cells in a dose-dependent fashion. Exodus-2 and -3 also induced dose-dependent chemotaxis of NKL, an IL-2-dependent human NK cell line. Results of modified checkerboard assays indicate that migration of NKL cells in response to Exodus-2 and -3 represents true chemotaxis and not simply chemokinesis. Exodus-1, -2, and -3 did not induce NK cell proliferation in the absence of other stimuli. Nevertheless, Exodus-2 and -3 significantly augmented IL-2-induced proliferation of normal human CD56(dim) NK cells. In contrast, Exodus-1, -2, and -3 did not affect the cytolytic activity of resting or activated peripheral blood NK cells. Expression of message for CCR7, a shared receptor for Exodus-2 and -3, was detected in activated polyclonal NK cells and NKL cells but not resting NK cells. Taken together, these results indicate that Exodus-2 and -3 can participate in the recruitment and proliferation of activated NK cells. Exodus-2 and -3 may regulate interactions between T cells and NK cells that are crucial for the generation of optimal immune responses. PMID- 10675271 TI - Human leptin enhances activation and proliferation of human circulating T lymphocytes. AB - Leptin is an adipocyte-secreted hormone that centrally regulates weight control. However, leptin receptor is expressed not only in the central nervous system, but also in other systems such as reproductive and hematopoietic tissues. Human leptin has previously been shown to enhance cytokine production by murine peritoneal macrophages and human circulating monocytes. In this paper we have assessed the presence of leptin receptors in peripheral human T lymphocytes and we have studied their functional role. Both CD4(+) and CD8(+) T lymphocytes express leptin receptors. Moreover, we show that human leptin dose-dependently enhances proliferation and activation of human circulating T lymphocytes when they are costimulated by PHA or Con A. Leptin alone was not able to activate T lymphocytes. To confirm a direct effect of leptin on T lymphocytes, monocytes were extracted by adhesion to culture flasks. The early activation surface marker CD69 was then induced in both CD4(+) and CD8(+) T lymphocytes after 8 h stimulation with PHA or Con A. Leptin dose-dependently enhanced stimulated CD69 expression. Moreover, leptin dose-dependently enhanced the expression of the late activation markers CD25 and CD71 in both CD4(+) and CD8(+) T lymphocytes after 48 h stimulation with PHA or Con A. Finally, we have found that leptin modulates CD4(+) T lymphocyte activation toward Th1 phenotype by stimulating the synthesis of IL-2 and IFN-gamma. These results demonstrate the presence of the leptin receptor in human circulating CD4(+) and CD8(+) T lymphocytes and a functional role of leptin as a modulator (enhancer) of lymphocyte stimulation with a shift toward Th1 cytokine-production profile. This function of leptin may have some relevance in the pathophysiology of immunologic alterations related to obesity. PMID- 10675272 TI - Synthetic melanin suppresses production of proinflammatory cytokines. AB - An overproduction of proinflammatory cytokines mediates the damaging sequelae of inflammation in pathologic conditions such as rheumatoid arthritis, graft-vs-host reaction, cachexia, and sepsis syndrome. We examined the cytokine regulatory activity of synthetic melanin, exemplified by biosynthetic l-glycine-l-tyrosine based polymer (ME-1) and chemosynthetic dihydroxyphenylalanine-based polymer (MC 1). At nontoxic concentrations, both compounds effectively (>/=60%) and reversibly suppressed the production of tumor necrosis factor (TNF), even when applied after stimulation of human peripheral blood monocytes with lipopolysaccharide (LPS). The inhibitory activity of melanin was selective with regard to cytokine response but not inducer- or cell-type-specific. In addition to TNF, melanin inhibited production of interleukin (IL)-1beta, IL-6, and IL-10 but not granulocyte-macrophage colony-stimulating factor by the LPS-stimulated monocytes. Melanin was equally effective in inhibiting production of TNF by monocytes stimulated with the purified protein derivative of Mycobacterium tuberculosis and production of IL-6 by IL-1alpha-stimulated human fibroblasts and endothelial cells. Northern blot analysis, mRNA stability determination, immunoprecipitation studies on metabolically labeled intracellular TNF, and pulse chase experiments revealed that melanin reduced efficiency of mRNA translation. The finding that melanin arrests ongoing cytokine synthesis suggests that this compound may be useful as an adjunct therapy for conditions showing involvement of proinflammatory cytokines. PMID- 10675273 TI - alpha-galactosylceramide induces early B-cell activation through IL-4 production by NKT cells. AB - alpha-Galactosylceramide (alpha-GalCer), a glycolipid antigen, specifically activates natural killer T (NKT) cells by a CD1d-restricted mechanism. In this work, we found that in vivo administration of alpha-GalCer resulted in the activation of B cells in addition to NKT cells, namely, alpha-GalCer administration caused upregulation of the early activation marker, CD69, on both NKT and B cells. In addition, expression of B7.2 and I-A(b) on B cells was greatly upregulated by alpha-GalCer. However, serum levels of IgE, IgG1, and IgG2a were not significantly changed within 48 h. In the present experiments, it was also demonstrated that the upregulation of CD69 expression by alpha-GalCer was strongly blocked by anti-IL-4 monoclonal antibody. Moreover, B-cell activation by alpha-GalCer was not observed in NKT-deficient mice. These results suggested that antigen-stimulated NKT cells might play a critical role not only in early defense mechanisms but also in early B-cell activation through IL-4 production. PMID- 10675274 TI - Macrophage-derived nitric oxide inhibits the proliferation of activated T helper cells and is induced during antigenic stimulation of resting T cells. AB - To examine how macrophage-derived nitric oxide (NO) affects T helper (Th) cell activity, T cell clones representing Th1 and Th2 subsets were activated before exposure to stimulated peritoneal macrophages or microglia. Both Th subsets were similarly sensitive to inhibition by NO, indicating that macrophage-derived NO regulates the proliferation of activated Th1 and Th2 cells equally well. Since IFN-gamma production remained intact in NO-treated Th1 cells, we studied whether NO was produced during antigen-specific activation of Th1 cells by unstimulated macrophages. Indeed, T cell proliferation only occurred when a NO synthase inhibitor was included, while IFN-gamma was essential for the induction of NO. These studies demonstrate that macrophages produce NO following antigen presentation to Th1 cells and that macrophage-derived NO inhibits Th1 and Th2 cell proliferation without inhibiting cytokine production. PMID- 10675275 TI - Induction of interleukin-12/p40 by superantigens in macrophages is mediated by activation of nuclear factor-kappaB. AB - Multimerization of the MHC class II molecule by superantigens results in activation of cellular signal transduction pathways in macrophage and B cells. Here we show that superantigen staphylococcal enterotoxin B (SEB) induces IL 12/p40 secretion in macrophages. SEB-induced expression of the IL-12/p40 gene involves activation and nuclear translocation of nuclear factor kappaB (NF kappaB). The NF-kappaB heterodimer bound to the NF-kappaB consensus sequence of the IL-12/p40 gene promoter is p50/C-Rel. Inhibition of PKC and PKA activation results in suppression of activation and translocation of NF-kappaB. We conclude that signals for IL-12/p40 gene transcription from MHC class II molecules follow activation of PKC and PKA, which in turn leads to the activation and translocation of NF-kappaB to the nucleus. Our study suggests that superantigens are capable of influencing the nature of the immune response by regulating cytokine production. Induction of IL-12 production by superantigens may therefore play a role in the regulation of Th 1-mediated immune response and autoimmune disease. PMID- 10675276 TI - Injection of plasmid DNA into the gastric mucosa induces mucosal and systemic immunity. AB - Nearly all mucosal surfaces participate in a common mucosal immune system, and application of an antigen to one mucosal surface elicits local as well as distant mucosal immune responses. However, whether the gastric mucosa is a part of this network has not been examined directly. We show here that the injection of plasmid DNA encoding beta-galactosidase into the gastric wall caused transfection of gastric mucosal epithelial cells, induced systemic and mucosal antibody responses at both local (digestive tract) and distant (genital and respiratory tracts) sites, and induced cytotoxic T lymphocyte responses in the spleen and the mesenteric and iliac lymph nodes. PMID- 10675277 TI - Is there a rationale for the use of creatine either as nutritional supplementation or drug administration in humans participating in a sport? AB - Even though no unambiguous proof for enhanced performance during high-intensity exercise has yet been reported, the creatine administration is charged to improve physical performance and has become a popular practice among subjects participating in different sports. Appropriate creatine dosage may be also used as a medicinal product since, in accordance with the Council Directive 65/65/CEE, any substance which may be administered with a view to restoring, correcting or modifying physiological functions in human beings is considered a medicinal product. Thus, quality, efficacy and safety must characterize the substance. In biochemical terms, creatine administration enhances both creatine and phosphocreatine concentrations, allowing for an increased total creatine pool in skeletal muscle. In thermodynamics terms, creatine interferes with the creatine creatine kinase-phosphocreatine circuit, which is related to the mitochondrial function as a highly organized system for the energy control of the subcellular adenylate pool. In pharmacokinetics terms, creatine entry into skeletal muscle is initially dependent on the extracellular concentration, but the creatine transport is subsequently down-regulated. In pharmacodynamics terms, the creatine enhances the possibility to maintain power output during brief periods of high intensity exercises. In spite of uncontrolled daily dosage and long-term administration, no research on creatine safety in humans has been set up by specific standard protocol of clinical pharmacology and toxicology, as currently occurs in phase I for the products for human use. More or less documented side effects induced by creatine are weight gain; influence on insulin production; feedback inhibition of endogenous creatine synthesis; long-term damages on renal function. A major point that related to the quality of creatine products is the amount of creatine ingested in relation to the amount of contaminants present. During the production of creatine from sarcosine and cyanamide, variable amounts of contaminants (dicyandiamide, dihydrotriazines, creatinine, ions) are generated and, thus, their tolerable concentrations (ppm) must be defined by specific toxicological researches. Creatine, as the nutritional factors, can be used either at supplementary or therapeutic levels as a function of the dose. Supplementary doses of nutritional factors usually are of the order of the daily turnover, while therapeutic ones are three or more times higher. In a subject with a body weight of 70 kg with a total creatine pool of 120 g, the daily turnover is approximately 2 g. Thus, in healthy subjects nourished with a fat rich, carbohydrate-, protein-poor diet and participating in a daily recreational sport, the oral creatine supplementation should be on the order of the daily turnover, i.e. less than 2.5-3 g per day, bringing the gastrointestinal absorption to account. In healthy athletes submitted daily to high-intensity strength- or sprint-training, the maximal oral creatine supplementation should be on the order of two times the daily turnover, i.e. less than 5-6 g per day for less than 2 weeks, and the creatine supplementation should be taken under appropriate medical supervision. The oral administration of more than 6 g per day of creatine should be considered as a therapeutic intervention because the dosage is more than three times higher than the creatine daily turnover and more than six times higher than the creatine daily allowance. In this case, creatine administration should be prescribed by physicians only in the cases of suspected or proven deficiency, or in conditions of severe stress and/or injury. 2000 Academic Press@p$hr PMID- 10675278 TI - Pepsinogens: physiology, pharmacology pathophysiology and exercise. AB - Human gastric mucosa contains aspartic proteinases that can be separated electrophoretically on the basis of their physical properties into two major groups: Pepsinogen I (PGA, PGI); and Pepsinogen II (PGC, PGII). Pepsinogens consist of a single polypeptide chain with molecular weight of approximately 42,000 Da. Pepsinogens are mainly synthesized and secreted by the gastric chief cells of the human stomach before being converted into the proteolytic enzyme pepsin, which is crucial for the digestive processes in the stomach. Pepsinogen synthesis and secretion are regulated by positive and negative feed-back mechanisms. In the resting state pepsinogens are stored in granules, which inhibit further synthesis. After appropriate physiological or external chemical stimuli, pepsinogens are secreted in the stomach lumen where hydrochloric acid, secreted by the parietal cells, converts them into the corresponding active enzyme pepsins. The stimulus-secreting coupling mechanisms of pepsinogens appear to include at least two major pathways: one involving cAMP as a mediator, the other involving modification of intracellular Ca(2+)concentration. Physiological or external chemical stimuli acting through the intracellular metabolic adenyl cyclase are more effective in inducing ' de novo ' pepsinogen synthesis than those acting through intracellular Ca(2+). The activation of protein kinase C (PK C) would appear to be involved in regulatory processes. The measurement of pepsinogens A and C in the serum is considered to be one of the non-invasive biochemical markers for monitoring peptic secretion and obtaining information on the gastric mucosa status of healthy subjects. Recently, pepsinogen measurements have been used as an effective biochemical method for evaluating and monitoring patients with gastrointestinal diseases and for checking the effects of drug treatment. The level of PGA in the serum is always high in normal gastritis, while in atrophic gastritis it is always low. In both cases the PGC level in the serum is high. In most gastrointestinal pathologies the ratio between the PGA/PGC decreases. Various reports concerning hormone and/or enzyme modification as well as gastrointestinal distress in the case of long distance exercise have been reported. It has been suggested that the origin of the gastrointestinal distress experienced by long distance runners is a transient ischaemia of the gastric mucosa; it is also suggested that a hypobaric-hypoxic environment could contribute to induce gastric mucosa necrosis. Interrelation between gastrointestinal distress, hypobaric-hypoxic environment and modifications of PGA and PGC, gastrin and cortisol was evaluated in 13 athletes after a marathon performed at 4300 m. Gastrointestinal symptoms occurred in approximately 40% of the athletes. After the race the athletes showed a significant increase of gastrin and cortisol, while the ratio between PGA/PGC decreased. No relationship was observed between gastrointestinal symptoms and hormonal changes after the race. A control group of five subjects, who had been exposed to the same environmental conditions, showed no gastrointestinal or hormonal alteration. Conversely, control subjects presented a significant decrease of cortisol related to the circadian rhythm. The same incidence of gastrointestinal symptoms at high altitude and at sea level and the absence of pathological alteration of PGA and PGC in the serum of the athletes indicates that running a marathon and living for 6 days at 4300 m does not induce gastric mucosa necrosis. Cortisol and gastrin alteration observed in the athletes at this altitude would seem to be related to an activation of the mesopontine and forebrain structures involved in the behavioural and metabolic integration of the autonomic control and arousal and psychophysical-exercise stress. 2000 Academic Press@p$hr PMID- 10675279 TI - Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: a possible mechanism of protection. AB - Administration of thymoquinone (10 mg kg(-1)day(-1), p.o.) with drinking water starting 5 days before a single injection of doxorubicin (15 mg kg(-1)i.p.) and continuing during the experimental period ameliorated the doxorubicin-induced cardiotoxicity in rats. This protection was evidenced from the significant reduction in serum enzymes: lactate dehydrogenase elevated level, 24 h and creatine phosphokinase elevated levels, 24 h and 48 h after doxorubicin administration. The cardiotoxicity of doxorubicin has been suggested to result from the generation of superoxide free-radical. The protective action of thymoquinone was examined against superoxide anion radical either generated photochemically, biochemically or derived from calcium ionophore (A23187) stimulated polymorphonuclear leukocytes. The results indicate that thymoquinone is a potent superoxide radical scavenger, scavenging power being as effective as superoxide dismutase against superoxide. In addition thymoquinone has an inhibitory effect on lipid peroxidation induced by Fe(3+)/ascorbate using rat heart homogenate. The superoxide scavenging and anti-lipid peroxidation may explain, in part, the protective effect of thymoquinone against doxorubicin induced cardiotoxicity. 2000 Academic Press@p$hr PMID- 10675280 TI - The potentiation of the histamine release induced by adenosine in mast cells from guinea pig lung and heart: sharp dependence on the time of preincubation. AB - We studied here the effect of a wide range of adenosine concentration and time of preincubation, on the histamine release induced in the guinea pig mast cells by different stimulus. Adenosine (10(-5)-10(-3)m) potentiated the histamine release induced by antigen in the guinea pig heart (isolated and dispersed tissue) and lung mast cells but not induced by ionophore A23197. The potentiation caused by adenosine (10(-4)m) was maximum after 1-3 min of preincubation and is probably an extracellular effect since it was not avoided by dipyridamol (3x10(-7)-10(-6)m) that inhibit the uptake of adenosine. Similar potentiation was also produced by the adenosine mimetic 2-chloroadenosine (10(-5)m) and both effects were inhibited by 8-phenyltheophylline indicating an effect on the type A receptors. It is suggested that the adenosine potentiation may not be related to changes on the cyclic AMP levels. 2000 Academic Press@p$hr PMID- 10675281 TI - Effect of dl-alpha-lipoic acid on the status of lipid peroxidation and antioxidants in aged rats. AB - The effect of dl-alpha-lipoic acid on lipid peroxidation and antioxidants status has been studied in the blood of young and aged rats. dl-alpha-lipoic acid, an antioxidant, was administered intraperitoneally for 7 and 14 days. Enzymatic and non-enzymatic antioxidant levels decreased with age but this decrease was attenuated by dl-alpha-lipoic acid. Lipid peroxide levels increased with age, and were decreased by lipoic acid administration. These results suggest that biochemical lesions which are considered to be part of the normal ageing process are neutralized by dl-alpha-lipoic acid. PMID- 10675282 TI - The effect of colchicine on proximal tubular reabsorption. AB - Aquaporins, expressed in the brush border membrane (BB) could play a pivotal role in glomerulo-tubular balance (GTB) by effecting adaptive changes of water permeability to the variations in the load of filtered solutes. Since aquaporin expression is modulated by microtubule-dependent trafficking between endoplasmic reticulum and cell membranes, we used the microtubule poison colchicine to assess the importance of aquaporins in mediating GTB. The effects of colchicine 1.6x10( 4)m on proximal tubule volume reabsorption was tested on 48 nephrons of ten rats by micropuncture techniques. Thirty proximal tubules were sampled from the last proximal convolution before, and recollected during and again after the microinjection (MIJ), into the early proximal convolution or Bowman's space, of colchicine added to a Ringer solution. We studied 18 proximal tubules in the same way before, during and after the microperfusion (MP) of colchicine added to an ultrafiltrate of plasma into the peritubular capillaries. During MIJ, SNGFR did not change significantly from baseline (17.7+/-1.3 vs 20.9+/-1.8 nl min(-1), P>0.12). Post-control values were superimposable upon their paired pre-MIJ controls, when available (15.8+/-1.3 vs 13.5+/-1.5 nl min(-1), P>0.25). The measurements of percentage reabsorption (49+/-5 during baseline, 45+/-7 during MIJ, and 55+/-5 in post-control, P>0.6) and absolute reabsorption (8.1+/-0.7, 11.1+/-2. 2, and 7.9+/-1.3 nl min(-1), respectively, P>0.18) were also unchanged. The three average measurements obtained in control conditions, during MP and again in post-MP control were not significantly different for SNGFR (19.8+/-3.0, 20.0+/-4.7, and 20. 2+/-3.5 nl min(-1), P>0.48), percentage (55+/-3, 59+/-5, and 47+/-3%, P>0.35) and absolute reabsorptions (12.5+/-2.2, 12.4+/-4.6, and 9. 4+/ 1.0 nl min(-1), respectively, P>0.42). MIJ and MP of vehicles were devoid of any measurable effect. Colchicine does not acutely affect volume reabsorption in the proximal tubule. Aquaporin trafficking, if any, is not involved in mediating glomerulotubular balance in the proximal tubule, although aquaporin expression and function could still be important, although regulated by mechanisms different from microtubule-dependent shuttling between endoplasmic reticulum and BB. PMID- 10675283 TI - Detection of anti-erythropoietin antibodies in haemodialysis patients treated with recombinant human-erythropoietin. AB - An enzyme-immunoassay was developed to evaluate the presence of anti erythropoietin antibodies in plasma samples obtained from renal failure patients treated with recombinant human erythropoietin (rh-EPO). The assay was specific and reproducible. Normal donors had no antibodies to EPO, while 67% of treated patients were positive to the assay. While the specificity of anti-EPO IgG antibodies was high, their affinity for the antigen was low. This finding can be explained by the very small differences in the structure of rh-EPO compared to that of natural EPO. The assay described could be useful in evaluating the long term effects of rh-EPO treatment on the control of anaemia in renal failure patients. PMID- 10675284 TI - Role of Moringa oleifera leaf extract in the regulation of thyroid hormone status in adult male and female rats. AB - The role of Moringa oleifera aqueous leaf extract in the regulation of thyroid hormone status, was studied in adult Swiss rats. Other than the thyroid hormone concentrations, hepatic lipid peroxidation (LPO) and the activities of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT) were evaluated. In the first experiment, effects of the leaf extract (175 mg kg( 1)body wt. day(-1)for 10 days) were studied both in male and female animals. Following the administration of the extract, serum triiodothyronine (T(3)) concentration and hepatic LPO decreased with a concomitant increase in the serum thyroxine (T(4)) concentration, in female rats, while in males no significant changes were observed, suggesting that Moringa oleifera leaf extract is more effective in females than in the males. To evaluate the impact of a higher dose, in the second experiment, the study was repeated in female rats, with 350 mg kg( 1)body wt. day(-1)for the same duration. Almost similar reduction in the serum T(3)concentration (approx. 30%) and an increase in the T(4)concentration were observed suggesting the inhibiting nature of Moringa oleifera leaf extract in the peripheral conversion of T(4)to T(3), the principal source of the generation of latter hormone. As the antiperoxidative effects were exhibited only by the lower dose and percent decrease in T(3)concentration was nearly the same by both the doses, it is suggested that the lower concentration of this plant extract may be used for the regulation of hyperthyriodism. 2000 Academic Press@p$hr PMID- 10675285 TI - Modulation of N-type calcium channels translocation in RINm5F insulinoma cells. AB - An intracellular pool of N-type voltage-operated calcium channels has recently been described in both IMR32 human neuroblastoma and PC12 rat pheochromocytoma cells. These channels were found to be accumulated in subcellular fractions where the chromogranin B-containing secretory granules were also enriched. Upon exocytosis N-type calcium channels were reversibly inserted in the plasma membrane. We have now extended this study to RINm5F rat insulinoma cells, and characterized the parallelism between the 'regulated' secretion of serotonin and the recruitment of surface calcium channels. Exocytosis was stimulated by different means, such as depolarization with high KCl, high Ba(2+)alone or protein kinase C activation; on the other hand exocytosis was inhibited with the non-selective calcium channel antagonist Cd(2+)or with noradrenaline. Stimulated release was always accompanied, with parallel kinetics, by calcium channel recruitment, while inhibition of secretion blocked calcium channel recruitment too. During repetitive depolarizations we revealed a potentiation of [Ca(2+)]()i transients in single Fura-2 loaded RINm5F cells, that was accompanied by an increase in surface VOCCs, suggesting a physiological role for the newly recruited channels. 2000 Academic Press@p$hr PMID- 10675286 TI - Inhibitory effect of KT3-671, a non-peptide angiotensin subtype 1 receptor antagonist, on sympathetic neurotransmission in isolated rabbit aorta. AB - Effect of KT3-671 on the sympathetic neurotransmission in isolated rabbit aorta was studied and compared with those of losartan and its active metabolite, EXP3174. Angiotensin (Ang) II (30 n m) produced approximately 1.7-fold increase in the transmural nerve stimulation (TNS)-evoked tritium overflow in the aorta preloaded with [(3)H]noradrenaline. KT3-671 (1 microm) by itself did not alter the TNS-evoked tritium overflow but it (0.1-1 microm) concentration-dependently inhibited the enhancing effect of Ang II on the TNS-evoked tritium overflow. Both losartan (1 and 3 microm) and EXP3174 (0.03-0.3 microm) also inhibited the Ang II effect. KT3-671 was approximately 8.6 and 0.3 times more potent than losartan and EXP3174, respectively, in inhibiting the Ang II response. This is consistent with the previous results showing the relative potency of the three antagonists to block AT(1)receptors. None of Ang II, KT3-671, losartan and EXP3174 affected significantly the spontaneous tritium outflow. These results suggest that KT3-671 as well as losartan and EXP3174 may inhibit vascular sympathetic neurotransmission by blocking presynaptic Ang II subtype 1 receptors, which appears to contribute partly to its antihypertensive action. 2000 Academic Press@p$hr PMID- 10675287 TI - Noradrenaline, dopamine, serotonin: different effects of psychological stress on brain biogenic amines in mice and rats. AB - The effect of restraint stress on central neurotransmission was evaluated in mice and rats. Noradrenaline (NA), dopamine (DA) and serotonin (5-HT) levels and their primary metabolites were measured in discrete brain regions following exposure to stress. Mice and rats demonstrated a similar response to stress in some brain regions. Both species responded to stress with lower NA and 5-HT in the locus coeruleus compared to non-stressed controls. Dopaminergic activity, assessed by DA turnover, was elevated in the hypothalamus. While DA turnover was suppressed in the amygdala, 5-HT turnover was similarly elevated in both species. In most cases, however, there were differences in biogenic neurotransmission between mice and rats in response to stress. In particular, NA levels were suppressed by stress in the dorsal cortex of mice, but in the rats NA levels were decreased in the hypothalamus. While stress produced lower DA levels in the hypothalamus, DA levels demonstrated a marked increase in the amygdala of mice. Stress was also associated with a decrease in DA levels in the rat striatum and with an increase of DA turnover in the locus coeruleus of mice. On the other hand, 5-HT was suppressed in the mouse striatum and in the rat hypothalamus and amygdala, while 5-HT turnover was markedly decreased in the hippocampus and dorsal cortex of rats alone. In conclusion, the changes in the central neurotransmission which are evoked by stress appear to be species-specific in most cases, a fact which may trigger discrete alterations in homeostatic mechanisms. PMID- 10675288 TI - Bilateral 6-OHDA lesions to the hippocampus attenuate the facilitatory effect of CCK-8 us and caerulein on memory in rats. AB - The involvement of dopaminergic projection to the hippocampus in the facilitatory effect of cholecystokinin-unsulphated octapeptide (CCK-8 us) and caerulein (CER) on memory motivated affectively was investigated in male rats. CCK-8 us and CER were given subcutaneously at the doses of 10 microg kg(-1)and 0.5 microg kg(-1), respectively, immediately after a single learning trial in a passive avoidance situation, after bilateral 6-OHDA lesions to the dentate gyrus of the hippocampus. In order to protect noradrenergic neurones against destruction by neurotoxin, 30 min before surgery rats were pre-treated intraperitoneally with 25 mg kg(-1)of desmethylimipramine, an inhibitor of noradrenaline uptake. Bilateral 6-OHDA lesions to the hippocampus significantly attenuate the facilitatory effect of CCK-8 us and CER on retention of passive avoidance behaviour evaluated 24 h after the learning trial. Neither, destruction of dopaminergic endings in the hippocampus, nor application of CCK-8 us and CER changed the spontaneous psychomotor activity of rats estimated in an 'open field' test. These results may indicate that the facilitatory effect of CCK-8 us and CER on memory motivated affectively is, in part, mediated by dopaminergic projection from the ventral tegmental area to the dentate gyrus of the hippocampus. PMID- 10675289 TI - Antinociceptive effect of amikacin and its interaction with morphine and naloxone. AB - Amikacin sulphate (30 mg kg(-1)) administered either intraperitoneally (i.p.) or subcutaneously (s.c.) produced antinociceptive effect in BALB/c mice in the acetic acid writhing test which is employed as an inflammatory pain model. The lack of difference between two routes with regard to antinociceptive potency was taken as evidence for the absence of a local effect. Amikacin sulphate-induced antinociception seems unlikely to be due to non-specific behaviour alteration, since this drug, at a dose range of 15-100 mg kg(-1)did not affect motor coordination of mice in rot-a-rod test. Morphine (1 mg kg(-1)) also caused antinociception when administered i.p. or s.c. but the effect was greater with the latter route. At the i.p. site; the concurrent use of amikacin and morphine produced more remarkable antinociception compared to their individual usages. Besides, naloxone (2 mg kg(-1)) significantly decreased antinociceptive effect of amikacin but itself also exerted antinociception. At present, we have no plausible explanation for these findings at the i.p. site. PMID- 10675290 TI - The hamster heart: a paradox in itself. AB - Perfusion of all mammalian heart muscle except hamster with Ca(2+)-free Tyrode and thereafter reperfusion with normal Tyrode causes irreversible damage, the calcium paradox. Our study aims at deciphering the role of creatine kinase, high energy phosphates and Ca(2+)influx in the genesis of myocardial injury in the rat and comparing it with the hamster. Isolated hearts from hamster and rats were perfused in the Langendorff mode at 37 degrees C for 30 min with normal Tyrode, for 15 min with Ca(2+)-free Tyrode and thereafter for 30 min of reperfusion with normal Tyrode. The 'high energy phosphate compound' levels were monitored by(31)P NMR, creatine kinase (CK) release was measured in the perfusate.(45)Ca influx was estimated in the papillary muscle. We observed that in the rat heart: (a) high energy phosphate levels declined significantly within 1 min of Ca(2+)reperfusion; (b) a massive release of CK occurred upon Ca(2+)reperfusion; (c) there was a significant increase of Ca(2+)influx. In the hamster heart, there was preservation of high energy phosphates, CK release was prevented completely and no rise in(45)Ca influx was observed upon Ca(2+)reperfusion. These results suggest that the hamster heart has a remarkable capacity for Ca(2+)homeostasis which protects the heart from Ca(2+)overload. PMID- 10675291 TI - Mechanism of the hypotensive action of Rhazya stricta leaf extract in rats. AB - The hypotensive action of Rhazya stricta lyophilized leaf extract was found to be partly caused by the electrolyte content of the extract, and partly caused by a strongly basic alkaloidal fraction (AF). AF (0.05-1.6 mg animal(-1)) caused a dose-dependent reduction in mean arterial blood pressure (MAP) of urethane anaesthetized rat preparations. In naiuml;ve pithed rats, AF administration (0.5 2.0 mg animal(-1)) significantly increased MAP. In pithed or spinalized rats made normotensive by noradrenaline infusion, AF (0.25 mg animal(-1)) did not cause any significant changes. Direct intracerebroventricular injection of AF (0.1-0.4 mg) markedly and significantly reduced MAP. It is suggested that the hypotensive action of AF to be mediated by a central mechanism. PMID- 10675292 TI - Three-dimensional structure of a vertebrate muscle Z-band: implications for titin and alpha-actinin binding. AB - The Z-band in vertebrate striated muscles, mainly comprising actin filaments, alpha-actinin, and titin, serves to organise the antiparallel actin filament arrays in adjacent sarcomeres and to transmit tension between sarcomeres during activation. Different Z-band thicknesses, formed from different numbers of zigzag crosslinking layers and found in different fibre types, are thought to be associated with the number of repetitive N-terminal sequence domains of titin. In order to understand myofibril formation it is necessary to correlate the ultrastructures and sequences of the actin filaments, titin, and alpha-actinin in characteristic Z-bands. Here electron micrographs of the intermediate width, basketweave Z-band of plaice fin muscle have been subject to a novel 3D reconstruction process. The reconstruction shows that antiparallel actin filaments overlap in the Z-band by about 22-25 nm. There are three levels of Z links (probably alpha-actinin) in which at each level two nearly diametrically opposed links join an actin filament to two of its antiparallel neighbours. One set of links is centrally located in the Z-band and there are flanking levels orthogonal to this. A 3D model of the observed structure shows how Z-bands of different widths may be formed and it provides insights into the structural arrangements of titin and alpha-actinin in the Z-band. The model shows that the two observed symmetries in different Z-bands, c2 and p12(1), may be attributed respectively to whether the number of Z-link levels is odd or even. PMID- 10675293 TI - Complementary visualization of mitotic barley chromatin by field-emission scanning electron microscopy and scanning force microscopy. AB - The surface structure of mitotic barley chromatin was studied by field-emission scanning electron microscopy (FESEM) and scanning force microscopy (SFM). Different stages of the cell cycle were accessible after a cell suspension was dropped onto a glass surface, chemical fixed, and critically point dried. Imaging was carried out with metal-coated specimen or uncoated specimen (only for SFM). The spatial contour of the chromatin could be resolved by SFM correlating to FESEM data. The experimentally determined volume of the residue chromatin during mitosis was within the range of 65-85 microm(3). A comparison with the theoretically calculated volume indicated a contribution of about 40% of internal cavities. Decondensation of chromosomes by proteinase K led to a drastic decrease in the chromosome volume, and a 3-D netlike architecture of the residue nucleoprotein material, similar to that in the intact chromosome, was obvious. Incubation of metaphase chromosomes in citrate buffer permitted access to different levels of chromatin packing. We imaged intact chromosomes in liquid by SFM without any intermediate drying step. A granular surface was obvious but with an appreciably lower resolution. Under similar imaging conditions proteinase K treated chromosomes exhibited low topographic contrast but were susceptible to plastic deformations. PMID- 10675294 TI - Structural organization of posterior midgut muscles in mosquitoes, Aedes aegypti and Anopheles gambiae. AB - In order to adapt to diverse feeding behavior, animal intestines have evolved with distinct differences. Such adaptation may include the structure of the longitudinal and circular muscles that maintain the integrity and the tensile strength of the gut tissue in higher metazoans. Here we examined the structural organization of the posterior midgut muscles of two insects, Aedes aegypti and Anopheles gambiae. We found the estimated number of longitudinal muscles in a cross-section to be 168 in Ae. aegypti and 37 in An. gambiae. Within the region, the estimated number of circular muscles is 77 in Ae. aegypti and 57 in An. gambiae. In An. gambiae, longitudinal muscles appear as sets of parallel bundles. Each set overlaps its neighbor to form a continuous tube. We found that this novel mode of muscle fiber sharing makes all circular muscles interconnected. Both types of muscle lie orthogonally to form a grid that holds the epithelium of the posterior midgut. In Ae. aegypti, the muscle fibers between the bundles are shared extensively, making the organization more intricate. This study implies that, because of its simple structure, the insect midgut may provide a powerful tool with which to study the structural evolution and function of animal intestines. PMID- 10675295 TI - Structure of abnormal molecular assemblies (collagen VI) associated with human full thickness macular holes. AB - Transversely banded deposits with an approximately 100-nm periodicity have been seen in association with a number of eye pathologies (e.g., age-related macular degeneration). Recently such aggregates have also been discovered in the cortical vitreous of a patient suffering from full thickness macular holes. The aggregates in the vitreous were of sufficient size and regularity for us to attempt 3D ultrastructural studies in the electron microscope. The molecules forming this aggregate pack in a centered tetragonal unit cell of dimensions approximately 26 x 26 x 180 nm. A real-space (r-weighted back projection) 3D reconstruction was computed. The aggregate is discussed in terms of its possible protein constituents. Collagen VI has been singled out as the most likely protein to form the aggregate. Two alternative models for the molecular packing are proposed, comprising aggregates of molecular tetramers or octamers. Understanding the structure of these abnormal banded deposits in the eye should help to throw light on the pathophysiological mechanisms of the diseases, including age-related macular degeneration, in which they occur. PMID- 10675296 TI - Cryo-electron tomography of neurospora mitochondria. AB - Cryo-electron tomography was used to study the structural organization of whole frozen-hydrated mitochondria from Neurospora crassa. Unlike mitochondria from many other species and tissues, in this case the cristae form a three-dimensional network of interconnected lamellae. Basically, the three-dimensional structure of ice-embedded mitochondria from this species is consistent with previous descriptions of mitochondria prepared by chemical fixation and resin embedding. Nonetheless, ice-embedded mitochondria display some important differences: the outer surface of the mitochondria was found to be rather smooth, the intermembrane space was constant in width, and distinct contact sites between the membranes were clearly revealed. Furthermore ATP synthase particles on the outer surface of an "inside-out vesicle" were visible in 3-D reconstructions. Thus, cryo-electron tomography can provide detailed insights into these organelles with minimal perturbations of the physiological state. This indicates that it is a realistic goal to achieve "molecular resolution" with rather large biological specimens in the near future, ultimately allowing the identification and localization of macromolecules in their cellular context. PMID- 10675297 TI - Quantitative microscopy of fluorescent adenovirus entry. AB - Fluorescence imaging of cells is a powerful tool for exploring the dynamics of organelles, proteins, and viruses. Fluorescent adenoviruses are a model system for cargo transport from the cell surface to the nucleus. Here, we describe a procedure to quantitate adenovirus-associated fluorescence in different subcellular regions. CCD camera-captured fluorescence sections across entire cells were deblurred by a fast Fourier transformation, the background was subtracted images merged, and virus fluorescence quantitated. The validity of the deblurring routine was verified by confocal laser scanning microscopy, demonstrating that objects were neither generated nor deleted. Instead, the homogeneity of both the average intensity and the size of fluorescent particles was increased, facilitating automated quantification. We found that nuclear fluorescence of wt adenovirus, but not of a virus mutant ts1, which fails to escape from endosomes, was maximal at 90 min postinfection (p.i.). Surprisingly, nuclear fluorescence decreased at 120 min, but increased again at 240 min p.i., suggesting that wt virus targeting to the nucleus may be multiphasic and regulated. Interestingly, only the first nuclear transport period of wt but not ts1 virus coincided with a significant increase of the peripheral and decrease of the cytoplasmic regions, indicative of signal-dependent cell contraction. PMID- 10675298 TI - The crystal structure of beta-glucosidase from Bacillus circulans sp. alkalophilus: ability to form long polymeric assemblies. AB - Family 1 of glycosyl hydrolases is a large and biologically important group of enzymes. A new three-dimensional structure of this family, beta-glucosidase from Bacillus circulans sp. alkalophilus is reported here. This is the first structure of beta-glucosidase from an alkaliphilic organism. The model was determined by the molecular replacement method and refined to a resolution of 2.7 A. The quaternary structure of B. circulans sp. alkalophilus beta-glucosidase is an octamer and subunits of the octamer show a similar (beta/alpha)(8) barrel fold to that previously reported for other family 1 enzymes. The crystal structure suggested that Cys169 in the active site is substituted. The Cys169 is located near the putative acid/base catalyst Glu166 and it may contribute to the high pH optimum of the enzyme. The crystal structure also revealed that the asymmetric unit contains two octamers which have a clear binding interaction with each other. The ability of the octamers to link with each other suggested that beta glucosidase from Bacillus circulans sp. alkalophilus is able to form long polymeric assemblies, at least in the crystalline state. PMID- 10675299 TI - Cryoelectron microscopy of protein-lipid complexes of human myelin basic protein charge isomers differing in degree of citrullination. AB - Myelin basic protein (MBP) is considered to be essential for the maintenance of stability of the myelin sheath. Reduction in cationicity of MBP, especially due to conversion of positively charged arginine residues to uncharged citrulline (Cit), has been found to be associated with multiple sclerosis (MS). Here, the interactions of an anionic phosphatidylserine/monosialoganglioside-G(M1) (4:1, w:w) lipid monolayer with 18.5-kDa MBP preparations from age-matched adult humans without MS (no Cit residues), with chronic MS (6 Cit), and with acute Marburg type MS (18 Cit) were studied by transmission and ultralow dose scanning transmission electron microscopy under cryogenic conditions. Immunogold labeling and single particle electron crystallography were used to define the nature of the complexes visualized. These electron microscopical analyses showed that the three different MBP charge isomers all formed uniformly sized and regularly shaped protein-lipid complexes with G(M1), probably as hexamers, but exhibited differential association with and organization of the lipid. The least cationic Marburg MBP-Cit(18) formed the most open protein-lipid complex. The data show a disturbance in lipid-MBP interactions at the ultrastructural level that is related to degree of citrullination, and which may be involved in myelin degeneration in multiple sclerosis. PMID- 10675300 TI - Crystallization and 1.1-A diffraction of chorismate lyase from Escherichia coli. AB - Chorismate pathway enzymes are important as producers of nonnucleotide aromatic compounds. The enzyme chorismate lyase from Escherichia coli has been crystallized in four distinct forms, three of which have been characterized by X ray diffraction. Despite widespread screening, all four crystal forms grow from the same chemical conditions. The wild-type enzyme tends to aggregate, even in the presence of reducing agent, and yielded only one crystal form (monoclinic, form 1) that grew in intricate clusters. Chemical modification of the cysteines mitigated problems with aggregation and solubility but did not affect crystal growth behavior. Protein aggregation was largely eliminated by mutating the protein's two cysteines to serines. The double mutant retains full enzymatic activity and crystallizes in three new forms, one of which (triclinic) diffracts to 1.1-A resolution. PMID- 10675301 TI - A unique Hawaiian Schiedea (caryophyllaceae: Alsinoideae) with only five fertile stamens. AB - The floral organogenesis and subsequent ontogenies of the Hawaiian endemic species Schiedea pubescens from Maui, Moloka'i, and Lana'i, and the Wai'anae Mountains, O'ahu, populations previously considered to be varietally distinct, were examined using the scanning electron microscope (SEM). The O'ahu population consistently produced only five fertile stamens, those of the inner whorl. The five stamens of the alternisepalous or outer whorl abort prior to microsporogenesis and fail to elongate. Additional vegetative differences between the two taxa, combined with the floral morphology characters, merit the description of the O'ahu population as a new species, S. pentandra, described herein. PMID- 10675302 TI - Comparative pollen morphology and ultrastructure of the Callitrichaceae. AB - The Callitrichaceae are an aquatic family of dicots that include the single, geographically cosmopolitan genus Callitriche. Callitriche contains 40-50 terrestrial, amphibious, and obligately submersed species, and it is the only known genus in the plant kingdom with co-occurring aerial and hydrophilous pollination syndromes. Pollen morphology and ultrastructure were described for 13 Callitriche species using scanning electron and transmission electron microscopy. Representative taxa of each growth form were examined; these included three terrestrial species (C. deflexa, C. peploides, and C. nuttallii), nine amphibious species (C. brutia, C. cophocarpa, C. cophocarpa-stagnalis hybrid, C. cribrosa, C. hamulata, C. heterophylla var. heterophylla, C. lusitanica, C. marginata, and C. trochlearis), and one obligately submersed species (C. truncata). Of the amphibious taxa, C. heterophylla var. heterophylla and C. trochlearis had internal geitonogamy, a type of internal self-fertilization. Pollen from all taxa was spheroidal, small, intectate, and lacked well-defined apertures. Taxa primarily differed with respect to exine thickness, surface ornamentation, and the presence or absence of aperture-like regions. The pollen of terrestrial species, as well as that of C. marginata, had well-developed exines with thick sculptured and basal layers. In general, amphibious taxa produced pollen with distinct, but thinner, exines than that of terrestrial taxa. Pollen of the amphibious taxa with internal geitonogamy had a thicker basal layer than species without internal geitonogamy, whereas the overall exine was reduced in C. hamulata and absent in C. brutia and C. lusitanica. Pollen of the obligately submersed C. truncata also lacked an exine. These palynological data were correlated with growth habits and related pollination biologies, as well as with phylogenetic interpretations of Callitrichaceae. Exine reduction or loss has evolved at least twice in the family, and it is associated with aneuploid reduction in chromosome number. PMID- 10675303 TI - SEM studies on vessels in ferns. 17. Psilotaceae. AB - Perforation plates are reported in aerial and subaerial axes of Psilotum nudum and in aerial axes of Tmesipteris obliqua. In Psilotum, both perforations lacking pit membranes and perforations with pit membrane remnants were observed. Perforation plates in Psilotum may consist wholly of one type or the other. In Tmespteris, perforations have threadlike pit membranes or consist of porose pit membranes. Wide perforations alternating with narrow pits, a conformation observed in various ferns, were observed in Psilotum (subaerial axes). In Psilotum, perforations are more common in metaxylem than in protoxylem; perforations in protoxylem consist of primary wall areas containing small circular porosities or relatively large circular to oval perforations. There are no modifications in the secondary wall framework of protoxylem or metaxylem in Psilotum or Tmesipteris that would permit one to distinguish presence of perforations or perforation plates with light microscopy, and scanning electron microscopy (SEM) is required for demonstration of porose walls or perforations. The tracheary elements of the Psilotaceae studied have no features not also observed in other ferns with SEM. PMID- 10675304 TI - Execution of the auxin replacement apical dominance experiment in temperate woody species. AB - The classic Thimann-Skoog or auxin replacement apical dominance test of exogenous auxin repression of lateral bud outgrowth was successfully executed in both seedlings and older trees of white ash, green ash, and red oak under the following conditions: (1) decapitation of a twig apex and auxin replacement were carried out during spring flush, (2) the decapitation was in the previous season's overwintered wood, and (3) the point of decapitation was below the upper large irrepressible lateral buds but above the lower repressible lateral buds. Although it has been suggested that neither auxin, the terminal bud, nor apical dominance have control over the outgrowth of the irrepressible buds during spring flush, there is evidence in the present study that indicates that such control over the repressible buds exists. In seedlings, second-order branching, which resulted from decapitation of elongating current shoots, was also inhibited by exogenous auxin in the three species. Hence, the auxin replacement experiments did work on year-old proleptic buds (of branches of older trees) that would have entered the bud bank and also on current buds of seedlings. Cytokinin treatments were ineffectual in promoting bud growth. PMID- 10675305 TI - The absence of cryptic self-incompatibility in Clarkia unguiculata (Onagraceae). AB - Many species exhibit reduced siring success of self-relative to outcross-pollen donors. This can be attributed either to postfertilization abortion of selfed ovules or to cryptic self-incompatibility (CSI). CSI is a form of self incompatibility whereby the advantage to outcross pollen is expressed only following pollinations where there is gametophytic competition between self and outcross pollen. Under the definition of CSI, this differential success is due to the superior prefertilization performance (pollen germination rate and pollen tube growth rate) of outcross pollen relative to self pollen. Although CSI has been demonstrated in several plant species, no studies have assessed among population variation in the expression of CSI. We conducted a greenhouse study on Clarkia unguiculata (an annual species with a mixed-mating system) to detect CSI, and we compare our observations to previous reports of CSI in C. gracilis and another population of C. unguiculata. In contrast to these previous studies of CSI in Clarkia, we used genetic rather than phenotypic markers to measure the relative performance of selfed vs. outcross pollen. In this study, we measured the intensity of CSI in C. unguiculata from a large population in southern California and we determined whether the magnitude of pollen competition (manipulated by controlling the number of pollen grains deposited on a stigma) influenced the outcome of competition between self and outcross pollen. In contrast to previous investigations of Clarkia, we found no evidence for CSI. The mean number of seeds sired per fruit did not differ between self and outcross pollen following either single-donor or mixed pollinations. In addition, the relative success of selfed vs. outcross pollen was independent of the magnitude of pollen competition. These results suggest that: (1) one of the few nonheterostylous species previously thought to be cryptically self-incompatible is completely self-compatible (at least in the population studied here) or (2) phenotypic markers may be problematic for the detection of CSI. PMID- 10675306 TI - Dissecting the causes of variation in intra-inflorescence allocation in a sexually polymorphic species, Fragaria virginiana (Rosaceae). AB - In this study we dissect the causes of variation in intra-inflorescence allocation in a sexually polymorphic species, Fragaria virginiana. We separated out the effects of resource competition during flowering from those of inflorescence architecture, as well as identified the effects of sex morph and genotype. We found position-based variation in petal length, ovule, pollen, and flower number to be influenced more by architecture than by our resource manipulations during flowering. We also found both genotype- and sex-specific intra-inflorescence patterns. Furthermore, our data indicate that the sex morph specific intra-inflorescence patterns result from architectural modifications of the basic pattern. In fact, sex-differential intra-inflorescence patterns suggest that fitness through male and female function may be maximized by different resource distribution patterns within the inflorescence and may have been modified by past selection. Specifically, females invested heavily in ovules at positions where fruit set was most likely (primary and secondary), at the expense of flower number and allocation per flower at more distal positions. Whereas functional males invested minimally in ovules at all flower positions and produced the most abundantly flowered inflorescences, hermaphrodites, on the other hand, showed intermediate patterns, implying a compromise between sex functions. We suggest that consideration of intra-inflorescence allocation and inflorescence architecture may reveal the mechanism underlying sexual dimorphism in flower allocation and number. PMID- 10675307 TI - Morphological variation and female reproductive success in two sympatric Trillium species: evidence for phenotypic selection in Trillium erectum and Trillium grandiflorum (Liliaceae). AB - I investigated the mating systems and phenotypic variation of two sympatric spring ephemerals, Trillium erectum and T. grandiflorum (Liliaceae), and phenotypic selection acting through female reproductive success for 11 morphological characters in five sympatric populations of the two species. I examined the degree of self-compatibility, pollinator-visitation rates, and pollen limitation of fruit and seed production in both species. Both Trillium species were self-compatible, but outcrossed flowers produced more successful fruits and seeds than self-pollinated flowers. Pollinator-visitation rates to the two species were low compared to other insect-pollinated spring ephemerals. In addition, both T. erectum and T. grandiflorum experienced pollen limitation in fruit and/or seed production; however, levels of fecundity in both species may be influenced by resource availability as well. I found significant phenotypic variation in 11 morphological characters within and among the five study populations. The sizes of all morphological characters were positively correlated. In general, larger T. erectum and T. grandiflorum produced more seeds. Phenotypic selection analysis revealed that direct and indirect selection acted on the size of morphological characters for both species. But there was no detectable selection acting on plant shape. This study reveals that variation in plant size exists within and among populations of both species, and this variation is associated with variance in female reproductive success. Spatial and temporal variation in pollinator and/or resource abundance may play a role in the phenotypic variation exhibited by both Trillium species. PMID- 10675308 TI - How accessible are receptive megastrobili to pollen? the example of jack pine (Pinus banksiana). AB - We examined the effects of wind speeds on pollen capture by megastrobili of jack pine (Pinus banksiana). We found that, when wind speed increased from 1.3 to 7.5 m/s, the relative capture efficiency (E(r)) did not change significantly (P <= 0.206) and remained below 12%. However, total capture rates increased linearly with wind speed and atmospheric pollen density. Because theoretical models of capture efficiency predict the E(r) to increase to ~80% asymptotically, our findings suggest that receptive megastrobili are equally adept at capturing pollen at all naturally occurring wind speeds. PMID- 10675309 TI - Biased sex ratios in the dioecious annual Croton texensis (Euphorbiaceae) are not due to environmental sex determination. AB - At Arapaho Prairie, in the sandhills of western Nebraska, the dioecious annual Croton texensis (Euphorbiaceae) exhibits biased sex ratios. Moreover, the direction of bias changes from year to year: in 1994 the study population was significantly female biased, in 1995 and 1996 it was significantly male biased, and in 1997 and 1998 the sex ratio did not differ from 1 : 1. Such variation in the observed sex ratio in plants is frequently attributed to environmental sex determination (ESD), which is favored by natural selection if the rate of fitness gain across an environmental gradient is greater for one sex than the other. We performed experiments to determine: (1) whether variation in the sex ratio is correlated with environmental conditions, as would be expected if ESD is operating, and (2) whether ESD, if present, would be favored by natural selection. In a common garden experiment in which water and fertilizer were manipulated the sex ratio was marginally male biased in treatments in which water was added, but not different from 1 : 1 in other treatments. In field plots into which seeds were planted none of several soil characteristics, nor overall plot quality for C. texensis (measured as average plant biomass) were correlated with plot sex ratio. However, plots in which a large number of planted seeds emerged tended to be female biased. These results provide very weak evidence for sex ratio bias across an environmental gradient, and thus provide little evidence for ESD. Moreover, sex-by-environment interactions for fitness, which are required for the evolution of ESD, were absent for all measured variables. Thus, ESD does not appear to be favored by natural selection in this population. Instead, these biases may have been caused by differences between the sexes in germination and/or early mortality. PMID- 10675310 TI - Patterns and determinants of potential carbon gain in the C3 evergreen Yucca glauca (Liliaceae) in a C4 grassland. AB - Yucca glauca is a C(3) evergreen rosette species locally common in the C(4) dominated grasslands of the central Great Plains. Most congeners of Y. glauca are found in deserts, and Y. glauca's morphological similarities to desert species (steeply angled leaves, evergreen habit) may be critical to its success in grasslands. We hypothesized that the evergreen habit of Y. glauca, coupled with its ability to remain physiologically active at cool temperatures, would allow this species to gain a substantial portion of its annual carbon budget when the C(4) grasses are dormant. Leaf-level gas exchange was measured over an 18-mo period at Konza Prairie in northeast Kansas to assess the annual pattern of potential C gain. Two short-term experiments also were conducted in which nighttime temperatures were manipulated to assess the cold tolerance of this species. The annual pattern of C gain in Y. glauca was bimodal, with a spring productive period (maximum monthly photosynthetic rate = 21.1 +/- 1.97 MUmol.m.s) in March through June, a period of midseason photosynthetic depression, and a fall productive period in October (15.6 +/- 1.25 MUmol.m.s). The steeply angled leaves resulted in interception of photon flux density at levels above photosynthetic saturation throughout the year. Reduced photosynthetic rates in the summer may have been caused by low soil moisture, but temperature was strongly related (r = 0.37) to annual variations in photosynthesis, with nocturnal air temperatures below -5 degrees C in the late fall and early spring, and high air temperatures (>32 degrees C) in the summer, limiting gas exchange. Overall, 31% of the potential annual carbon gain in Y. glauca occurred outside the "frost-free" period (April-October) at Konza Prairie and 43% occurred when the dominant C(4) grasses were dormant. Future climates that include warmer minimum temperatures in the spring and fall may enhance the success of Y. glauca relative to the C(4) dominants in these grasslands. PMID- 10675311 TI - Clonal growth of Lithospermum caroliniense (Boraginaceae) in contrasting sand dune habitats. AB - The occurrence of clonal growth of distylous Lithospermum caroliniense was investigated in a population in the Nebraska Sandhills, an area where sand dunes have been relatively stable for at least 1500-3000 yr, and compared to a population occurring at the Indiana Dunes, an area of active sand dune formation. Spatial autocorrelation analysis indicated the occurrence of significant clonal propagation of genetically based floral morphs at Arapaho Prairie, but not for the Indiana Dunes. Apparent clonal growth in the Sandhills population had no overall negative effect on pollen deposition or fecundity relative to the Indiana population, although in some large clones the proportion of compatible pollen grains on stigmas was lower. Clonal growth may have occurred in the Sandhills population because of the greater age and stability of the Nebraska Sandhills; infrequent establishment of seedlings permits detection of clonal growth using the spatial pattern of floral morphs. At the Indiana dunes, repeated cycles of dune formation provide conditions favoring establishment of seedlings, and sand dune succession results in disappearance of L. caroliniense before the development of clones. PMID- 10675312 TI - Interactive effects of elevated CO2 and temperature on water transport inponderosa pine. AB - Many studies report that water flux through trees declines in response to elevated CO(2), but this response may be modified by exposure to increased temperatures. To determine whether elevated CO(2) and temperature interact to affect hydraulic conductivity, we grew ponderosa pine seedlings for 24 wk in growth chambers with one of four atmospheric CO(2) concentrations (350, 550, 750, and 1100 ppm) and either a low (15 degrees C nights, 25 degrees C days) or high (20 degrees C nights, 30 degrees C days) temperature treatment. Vapor pressure deficits were also higher in the elevated temperature treatment. Seedling biomass increased with CO(2) concentration but was not affected by temperature. Root : shoot ratio was unaffected by CO(2) and temperature. Leaf : sapwood area ratio (A(L)/A(S)) declined in response to elevated temperature but was not influenced by CO(2). Larger tracheid diameters at elevated temperature caused an increase in xylem-specific hydraulic conductivity (K(S)). The increase in K(S) and decrease in A(L)/A(S) led to higher leaf-specific hydraulic conductivity (K(L)) at elevated temperature. Stomatal conductance (g(S)) was correlated with K(L) across all treatments. Neither K(S), K(L), nor g(S) were affected by elevated CO(2) concentrations. High K(L) in response to elevated temperature may support increased transpiration or reduce the incidence of xylem cavitation in ponderosa pine in future, warmer climates. PMID- 10675313 TI - Genetic variation in chloroplast and nuclear ribosomal DNA in Utah juniper (Juniperus osteosperma, Cupressaceae): evidence for interspecific gene flow. AB - Geographic patterns of genetic variation in chlorolast (cpDNA) and nuclear ribosomal (nrDNA) DNA were examined to test the hypothesis of hybridization between Juniperus osteosperma and Juniperus occidentalis in the Great Basin of western Nevada. Noncoding DNA from the trnL-trnF intergenic spacer and the trnL intron of the chloroplast genome was sequenced from seven populations of J. osteosperma and four populations of J. occidentalis sampled over a large proportion of their respective ranges. An adenine nucleotide at position 436 in the aligned sequence and within a Tru 9I restriction site was found to be present in individuals of J. osteosperma sampled from western Colorado and central Utah, but absent in sequences of J. osteosperma sampled from central and western Nevada and all sequences of J. occidentalis. Two hundred fourteen individuals from 34 populations of J. osteosperma and J. occidentalis were then screened for cpDNA haplotype by Tru 9I digestion of the trnL-trnF polymerase chain reaction (PCR) product. Two cpDNA haplotypes were evident, each consisting of restriction fragment profiles that differed solely with respect to the presence or absence of the Tru 9I site encompassing the adenine nucleotide at position 436. One of these haplotypes was monomorphic in J. occidentalis and exhibited a decreasing frequency in J. osteosperma with increasing geographic distance from J. occidentalis in west-central Nevada. Geographic patterns in nuclear ribosomal DNA (nrDNA) variation were examined by restriction fragment analysis and, although spatially more restricted, exhibited patterns of clinal variation similar to those observed in cpDNA haplotype. Genetic relationships based on DNA sequences and geographic patterns of genetic variation in chloroplast and nuclear ribosomal DNA are consistent with morphology in suggesting interspecific gene flow between J. occidentalis and J. osteosperma. PMID- 10675314 TI - Phylogeny of South African Gnaphalieae (Asteraceae) based on two noncoding chloroplast sequences. AB - The Gnaphalieae are a group of sunflowers that have their greatest diversity in South America, Southern Africa, and Australia. The objective of this study was to reconstruct a phylogeny of the South African Gnaphalieae using sequence data from two noncoding chloroplast DNA sequences, the trnL intron and trnL/trnF intergenic spacer. Included in this investigation are the genera of the Gnaphalieae from the African basal groups, members of the subtribes Cassiniinae, Gnaphaliinae, and Relhaniinae, and African representatives from the large Old World genus Helichrysum. Results indicate that two Gnaphaloid genera, Printzia and Callilepis, should be excluded from the Gnaphalieae. In most trees the Relhaniinae s.s. (sensu stricto) and some of the basal taxa comprise a clade that is sister to the remainder of the tribe Gnaphalieae. The Relhaniinae, which are restricted to Africa, are not a monophyletic group as presently circumscribed, nor are the South African members of Helichrysum, the Cassiniinae and Gnaphaliinae. There is general agreement between our molecular analysis and that of morphology, particularly in the terminal branches of the trees. PMID- 10675315 TI - A phylogeny of the flowering plant family Apiaceae based on chloroplast DNA rpl16 and rpoC1 intron sequences: towards a suprageneric classification of subfamily Apioideae. AB - The higher level relationships within Apiaceae (Umbelliferae) subfamily Apioideae are controversial, with no widely acceptable modern classification available. Comparative sequencing of the intron in chloroplast ribosomal protein gene rpl16 was carried out in order to examine evolutionary relationships among 119 species (99 genera) of subfamily Apioideae and 28 species from Apiaceae subfamilies Saniculoideae and Hydrocotyloideae, and putatively allied families Araliaceae and Pittosporaceae. Phylogenetic analyses of these intron sequences alone, or in conjunction with plastid rpoC1 intron sequences for a subset of the taxa, using maximum parsimony and neighbor-joining methods, reveal a pattern of relationships within Apioideae consistent with previously published chloroplast DNA and nuclear ribosomal DNA ITS based phylogenies. Based on consensus of relationship, seven major lineages within the subfamily are recognized at the tribal level. These are referred to as tribes Heteromorpheae M. F. Watson & S. R. Downie Trib. Nov., Bupleureae Spreng. (1820), Oenantheae Dumort. (1827), Pleurospermeae M. F. Watson & S. R. Downie Trib. Nov., Smyrnieae Spreng. (1820), Aciphylleae M. F. Watson & S. R. Downie Trib. Nov., and Scandiceae Spreng. (1820). Scandiceae comprises subtribes Daucinae Dumort. (1827), Scandicinae Tausch (1834), and Torilidinae Dumort. (1827). Rpl16 intron sequences provide valuable characters for inferring high-level relationships within Apiaceae but, like the rpoC1 intron, are insufficient to resolve relationships among closely related taxa. PMID- 10675316 TI - Protein phosphorylation and protein phosphatases. De Panne, Belgium, September 19 24, 1999. PMID- 10675317 TI - Macromolecular mimicry. AB - Some proteins have been shown to mimic the overall shape and structure of nucleic acids. For some of the proteins involved in translating the genetic information into proteins on the ribosome particle, there are indications that such observations of macromolecular mimicry even extend to similarity in interaction with and function on the ribosome. A small number of structural results obtained outside the protein biosynthesis machinery could indicate that the concept of macromolecular mimicry between proteins and nucleic acids is more general. The implications for the function and evolution of protein biosynthesis are discussed. PMID- 10675318 TI - Multiple pathways control protein kinase C phosphorylation. PMID- 10675319 TI - Crystal structure of human sex hormone-binding globulin: steroid transport by a laminin G-like domain. AB - Human sex hormone-binding globulin (SHBG) transports sex steroids in blood and regulates their access to target tissues. In biological fluids, SHBG exists as a homodimer and each monomer comprises two laminin G-like domains (G domains). The crystal structure of the N-terminal G domain of SHBG in complex with 5alpha dihydrotestosterone at 1.55 A resolution reveals both the architecture of the steroid-binding site and the quaternary structure of the dimer. We also show that G domains have jellyroll topology and are structurally related to pentraxin. In each SHBG monomer, the steroid intercalates into a hydrophobic pocket within the beta-sheet sandwich. The steroid and a 20 A distant calcium ion are not located at the dimer interface. Instead, two separate steroid-binding pockets and calcium binding sites exist per dimer. The structure displays intriguing disorder for loop segment Pro130-Arg135. In all other jellyroll proteins, this loop is well ordered. If modelled accordingly, it covers the steroid-binding site and could thereby regulate access of ligands to the binding pocket. PMID- 10675320 TI - B cells extract and present immobilized antigen: implications for affinity discrimination. AB - Binding of antigen to B-cell antigen receptor (BCR) leads to antigen internalization and presentation to T cells, a critical process in the initiation of the humoral immune response. However, antigen internalization has been demonstrated for soluble antigen, in vivo antigen is often encountered in insoluble form or tethered to a cell surface. Here, we show that not only can B cells internalize and present large particulate antigen (requiring a signalling competent BCR to drive antigen uptake), but they can also extract antigen that is tethered tightly to a non-internalizable surface. The form in which the antigen is displayed affects the B cell's ability to discriminate antigen-BCR affinity. Thus, arraying an antigen on a particle or surface allows efficient presentation of low affinity antigens. However, the presentation efficiency of antigen arrayed on an internalizable particle plateaus at low affinity values. In contrast, extraction and presentation of antigen from a non-internalizable surface depends on antigen-BCR affinity over a wide affinity range. The results have implications for understanding both the initiation and affinity maturation of the immune response. PMID- 10675321 TI - Activation of rho through a cross-link with polyamines catalyzed by Bordetella dermonecrotizing toxin. AB - The small GTPase Rho, which regulates a variety of cell functions, also serves as a specific substrate for bacterial toxins. Here we demonstrate that Bordetella dermonecrotizing toxin (DNT) catalyzes cross-linking of Rho with ubiquitous polyamines such as putrescine, spermidine and spermine. Mass spectrometric analyses revealed that the cross-link occurred at Gln63, which had been reported to be deamidated by DNT in the absence of polyamines. Rac1 and Cdc42, other members of the Rho family GTPases, were also polyaminated by DNT. The polyamination, like the deamidation, markedly reduced the GTPase activity of Rho without affecting its GTP-binding activity, indicating that polyaminated Rho behaves as a constitutively active analog. Moreover, polyamine-linked Rho, even in the GDP-bound form, associated more effectively with its effector ROCK than deamidated Rho in the GTP-bound form and, when microinjected into cells, induced the anomalous formation of stress fibers indistinguishable from those seen in DNT treated cells. The results imply that the polyamine-linked Rho, transducing signals to downstream ROCK in a novel GTP-independent manner, plays an important role in DNT cell toxicity. PMID- 10675322 TI - Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity. AB - FtsY, the Escherichia coli homologue of the eukaryotic signal recognition particle (SRP) receptor alpha-subunit, is located in both the cytoplasm and inner membrane. It has been proposed that FtsY has a direct targeting function, but the mechanism of its association with the membrane is unclear. FtsY is composed of two hydrophilic domains: a highly charged N-terminal domain (the A-domain) and a C-terminal GTP-binding domain (the NG-domain). FtsY does not contain any hydrophobic sequence that might explain its affinity for the inner membrane, and a membrane-anchoring protein has not been detected. In this study, we provide evidence that FtsY interacts directly with E.coli phospholipids, with a preference for anionic phospholipids. The interaction involves at least two lipid binding sites, one of which is present in the NG-domain. Lipid association induced a conformational change in FtsY and greatly enhanced its GTPase activity. We propose that lipid binding of FtsY is important for the regulation of SRP mediated protein targeting. PMID- 10675323 TI - YidC, the Escherichia coli homologue of mitochondrial Oxa1p, is a component of the Sec translocase. AB - In Escherichia coli, both secretory and inner membrane proteins initially are targeted to the core SecYEG inner membrane translocase. Previous work has also identified the peripherally associated SecA protein as well as the SecD, SecF and YajC inner membrane proteins as components of the translocase. Here, we use a cross-linking approach to show that hydrophilic portions of a co-translationally targeted inner membrane protein (FtsQ) are close to SecA and SecY, suggesting that insertion takes place at the SecA/Y interface. The hydrophobic FtsQ signal anchor sequence contacts both lipids and a novel 60 kDa translocase-associated component that we identify as YidC. YidC is homologous to Saccharomyces cerevisiae Oxa1p, which has been shown to function in a novel export pathway at the mitochondrial inner membrane. We propose that YidC is involved in the insertion of hydrophobic sequences into the lipid bilayer after initial recognition by the SecAYEG translocase. PMID- 10675324 TI - Degradation of unassembled Vph1p reveals novel aspects of the yeast ER quality control system. AB - The endoplasmic reticulum quality control (ERQC) system retains and degrades soluble and membrane proteins that misfold or fail to assemble. Vph1p is the 100 kDa membrane subunit of the yeast Saccharomyces cerevisiae V-ATPase, which together with other subunits, assembles into the V-ATPase in the ER, requiring the ER resident protein Vma22p. In vma22Delta cells, Vph1p remains an integral membrane protein with wild-type topology in the ER membrane before undergoing a rapid and concerted degradation requiring neither vacuolar proteases nor transport to the Golgi. Failure to assemble targets Vph1p for degradation in a process involving ubiquitylation, the proteasome and cytosolic but not ER lumenal chaperones. Vph1p appears to possess the traits of a 'classical' ERQC substrate, yet novel characteristics are involved in its degradation: (i) UBC genes other than UBC6 and UBC7 are involved and (ii) components of the ERQC system identified to date (Der1p, Hrd1p/Der3p and Hrd3p) are not required. These data suggest that other ERQC components must exist to effect the degradation of Vph1p, perhaps comprising an alternative pathway. PMID- 10675325 TI - A truncated isoform of the PP2A B56 subunit promotes cell motility through paxillin phosphorylation. AB - Both F10 and BL6 sublines of B16 mouse melanoma cells are metastatic after intravenous injection, but only BL6 cells are metastatic after subcutaneous injection. Retrotransposon insertion was found to produce an N-terminally truncated form (Deltagamma1) of the B56gamma1 regulatory subunit isoform of protein phosphatase (PP) 2A in BL6 cells, but not in F10 cells. We found an interaction of paxillin with PP2A C and B56gamma subunits by co immunoprecipitation. B56gamma1 co-localized with paxillin at focal adhesions, suggesting a role for this isoform in targeting PP2A to paxillin. In this regard, Deltagamma1 behaved similarly to B56gamma1. However, the Deltagamma1-containing PP2A heterotrimer was insufficient for the dephosphorylation of paxillin. Transfection with Deltagamma1 enhanced paxillin phosphorylation on serine residues and recruitment into focal adhesions, and cell spreading with an actin network. In addition, Deltagamma1 rendered F10 cells as highly metastatic as BL6 cells. These results suggest that mutations in PP2A regulatory subunits may cause malignant progression. PMID- 10675326 TI - Op18/stathmin caps a kinked protofilament-like tubulin tetramer. AB - Oncoprotein 18/stathmin (Op18), a regulator of microtubule dynamics, was recombinantly expressed and its structure and function analysed. We report that Op18 by itself can fold into a flexible and extended alpha-helix, which is in equilibrium with a less ordered structure. In complex with tubulin, however, all except the last seven C-terminal residues of Op18 are tightly bound to tubulin. Digital image analysis of Op18:tubulin electron micrographs revealed that the complex consists of two longitudinally aligned alpha/beta-tubulin heterodimers. The appearance of the complex was that of a kinked protofilament-like structure with a flat and a ribbed side. Deletion mapping of Op18 further demonstrated that (i) the function of the N-terminal part of the molecule is to 'cap' tubulin subunits to ensure the specificity of the complex and (ii) the complete C terminal alpha-helical domain of Op18 is necessary and sufficient for stable Op18:tubulin complex formation. Together, our results suggest that besides sequestering tubulin, the structural features of Op18 enable the protein specifically to recognize microtubule ends to trigger catastrophes. PMID- 10675327 TI - Crystal structure of a class I alpha1,2-mannosidase involved in N-glycan processing and endoplasmic reticulum quality control. AB - Mannose trimming is not only essential for N-glycan maturation in mammalian cells but also triggers degradation of misfolded glycoproteins. The crystal structure of the class I alpha1, 2-mannosidase that trims Man(9)GlcNAc(2) to Man(8)GlcNAc(2 )isomer B in the endoplasmic reticulum of Saccharomyces cerevisiae reveals a novel (alphaalpha)(7)-barrel in which an N-glycan from one molecule extends into the barrel of an adjacent molecule, interacting with the essential acidic residues and calcium ion. The observed protein-carbohydrate interactions provide the first insight into the catalytic mechanism and specificity of this eukaryotic enzyme family and may be used to design inhibitors that prevent degradation of misfolded glycoproteins in genetic diseases. PMID- 10675328 TI - Induction of apoptosis by Drosophila reaper, hid and grim through inhibition of IAP function. AB - Induction of apoptosis in Drosophila requires the activity of three closely linked genes, reaper, hid and grim. Here we show that the proteins encoded by reaper, hid and grim activate cell death by inhibiting the anti-apoptotic activity of the Drosophila IAP1 (diap1) protein. In a genetic modifier screen, both loss-of-function and gain-of-function alleles in the endogenous diap1 gene were obtained, and the mutant proteins were functionally and biochemically characterized. Gain-of-function mutations in diap1 strongly suppressed reaper-, hid- and grim-induced apoptosis. Sequence analysis of these alleles revealed that they were caused by single amino acid changes in the baculovirus IAP repeat domains of diap1, a domain implicated in binding REAPER, HID and GRIM. Significantly, the corresponding mutant DIAP1 proteins displayed greatly reduced binding of REAPER, HID and GRIM, indicating that REAPER, HID and GRIM kill by forming a complex with DIAP1. These data provide strong in vivo evidence for a previously published model of cell death regulation in Drosophila. PMID- 10675329 TI - The Drosophila caspase DRONC is regulated by DIAP1. AB - We have isolated the recently identified Drosophila caspase DRONC through its interaction with the effector caspase drICE. Ectopic expression of DRONC induces cell death in Schizosaccharomyces pombe, mammalian fibroblasts and the developing Drosophila eye. The caspase inhibitor p35 fails to rescue DRONC-induced cell death in vivo and is not cleaved by DRONC in vitro, making DRONC the first identified p35-resistant caspase. The DRONC pro-domain interacts with Drosphila inhibitor of apoptosis protein 1 (DIAP1), and co-expression of DIAP1 in the developing Drosophila eye completely reverts the eye ablation phenotype induced by pro-DRONC expression. In contrast, DIAP1 fails to rescue eye ablation induced by DRONC lacking the pro-domain, indicating that interaction of DIAP1 with the pro-domain of DRONC is required for suppression of DRONC-mediated cell death. Heterozygosity at the diap1 locus enhances the pro-DRONC eye phenotype, consistent with a role for endogenous DIAP1 in suppression of DRONC activation. Both heterozygosity at the dronc locus and expression of dominant-negative DRONC mutants suppress the eye phenotype caused by reaper (RPR) and head involution defective (HID), consistent with the idea that DRONC functions in the RPR and HID pathway. PMID- 10675330 TI - C-cell hyperplasia, pheochromocytoma and sympathoadrenal malformation in a mouse model of multiple endocrine neoplasia type 2B. AB - Dominantly inherited multiple endocrine neoplasia type 2B (MEN2B) is characterized by tumors of the thyroid C-cells and adrenal chromaffin cells, together with ganglioneuromas of the gastrointestinal tract and other developmental abnormalities. Most cases are caused by substitution of threonine for Met918 in the RET receptor tyrosine kinase, which is believed to convert the RET gene to an oncogene by altering the enzyme's substrate specificity. We report the production of a mouse model of MEN2B by introduction of the corresponding mutation into the ret gene. Mutant mice displayed C-cell hyperplasia and chromaffin cell hyperplasia progressing to pheochromocytoma. Homozygotes did not develop gastrointestinal ganglioneuromas, but displayed ganglioneuromas of the adrenal medulla, enlargement of the associated sympathetic ganglia and a male reproductive defect. Surprisingly, homozygotes did not display any developmental defects attributable to a loss-of-function mutation. Thus, while our results support the conclusion that the Met918Thr substitution is responsible for MEN2B, they suggest that the substrate specificity of the RET kinase does not interfere with its normal role in the development of the kidneys and enteric nervous system. PMID- 10675331 TI - An EGF receptor/Ral-GTPase signaling cascade regulates c-Src activity and substrate specificity. AB - c-Src is a membrane-associated tyrosine kinase that can be activated by many types of extracellular signals, and can regulate the function of a variety of cellular protein substrates. We demonstrate that epidermal growth factor (EGF) and beta-adrenergic receptors activate c-Src by different mechanisms leading to the phosphorylation of distinct sets of c-Src substrates. In particular, we found that EGF receptors, but not beta(2)-adrenergic receptors, activated c-Src by a Ral-GTPase-dependent mechanism. Also, c-Src activated by EGF treatment or expression of constitutively activated Ral-GTPase led to tyrosine phosphorylation of Stat3 and cortactin, but not Shc or subsequent Erk activation. In contrast, c Src activated by isoproterenol led to tyrosine phosphorylation of Shc and subsequent Erk activation, but not tyrosine phosphorylation of cortactin or Stat3. These results identify a role for Ral-GTPases in the activation of c-Src by EGF receptors and the coupling of EGF to transcription through Stat3 and the actin cytoskeleton through cortactin. They also show that c-Src kinase activity can be used differently by individual extracellular stimuli, possibly contributing to their ability to generate unique cellular responses. PMID- 10675332 TI - N-acyl homoserine lactone binding to the CarR receptor determines quorum-sensing specificity in Erwinia. AB - Quorum sensing via an N-acyl homoserine lactone (HSL) pheromone controls the biosynthesis of a carbapenem antibiotic in Erwinia carotovora. Transcription of the carbapenem biosynthetic genes is dependent on the LuxR-type activator protein, CarR. Equilibrium binding of a range of HSL molecules, which are thought to activate CarR to bind to its DNA target sequence, was examined using fluorescence quenching, DNA bandshift analysis, limited proteolysis and reporter gene assays. CarR bound the most physiologically relevant ligand, N-(3 oxohexanoyl)-L-homoserine lactone, with a stoichiometry of two molecules of ligand per dimer of protein and a dissociation constant of 1.8 microM, in good agreement with the concentration of HSL required to activate carbapenem production in vivo. In the presence of HSL, CarR formed a very high molecular weight complex with its target DNA, indicating that the ligand causes the protein to multimerize. Chemical cross-linking analysis supported this interpretation. Our data show that the ability of a given HSL to facilitate CarR binding to its target DNA sequence is directly proportional to the affinity of the HSL for the protein. PMID- 10675333 TI - The Sos1 and Sos2 Ras-specific exchange factors: differences in placental expression and signaling properties. AB - Targeted disruption of both alleles of mouse sos1, which encodes a Ras-specific exchange factor, conferred mid-gestational embryonic lethality that was secondary to impaired placental development and was associated with very low placental ERK activity. The trophoblastic layers of sos1(-/-) embryos were poorly developed, correlating with high sos1 expression in wild-type trophoblasts. A sos1(-/-) cell line, which expressed readily detectable levels of the closely related Sos2 protein, formed complexes between Sos2, epidermal growth factor receptor (EGFR) and Shc efficiently, gave normal Ras.GTP and ERK responses when treated with EGF for < or =10 min and was transformed readily by activated Ras. However, the sos1( /-) cells were resistant to transformation by v-Src or by overexpressed EGFR and continuous EGF treatment, unlike sos1(+/-) or wild-type cells. This correlated with Sos2 binding less efficiently than Sos1 to EGFR and Shc in cells treated with EGF for > or =90 min or to v-Src and Shc in v-Src-expressing cells, and with less ERK activity. We conclude that Sos1 participates in both short- and long term signaling, while Sos2-dependent signals are predominantly short-term. PMID- 10675334 TI - Transcription factor Sp3 is essential for post-natal survival and late tooth development. AB - Sp3 is a ubiquitously expressed transcription factor closely related to Sp1 (specificity protein 1). We have disrupted the mouse Sp3 gene by homologous recombination. Sp3-deficient embryos are growth retarded and invariably die at birth of respiratory failure. The cause for the observed breathing defect remains obscure since only minor morphological alterations were observed in the lung, and surfactant protein expression is indistinguishable from that in wild-type mice. Histological examinations of individual organs in Sp3(-/-) mice show a pronounced defect in late tooth formation. In Sp3 null mice, the dentin/enamel layer of the developing teeth is impaired due to the lack of ameloblast-specific gene products. Comparison of the Sp1 and Sp3 knockout phenotype shows that Sp1 and Sp3 have distinct functions in vivo, but also suggests a degree of functional redundancy. PMID- 10675335 TI - Regulation of E2F1 activity by acetylation. AB - During the G(1) phase of the cell cycle, an E2F-RB complex represses transcription, via the recruitment of histone deacetylase activity. Phosphorylation of RB at the G(1)/S boundary generates a pool of 'free' E2F, which then stimulates transcription of S-phase genes. Given that E2F1 activity is stimulated by p300/CBP acetylase and repressed by an RB-associated deacetylase, we asked if E2F1 was subject to modification by acetylation. We show that the p300/CBP-associated factor P/CAF, and to a lesser extent p300/CBP itself, can acetylate E2F1 in vitro and that intracellular E2F1 is acetylated. The acetylation sites lie adjacent to the E2F1 DNA-binding domain and involve lysine residues highly conserved in E2F1, 2 and 3. Acetylation by P/CAF has three functional consequences on E2F1 activity: increased DNA-binding ability, activation potential and protein half-life. These results suggest that acetylation stimulates the functions of the non-RB bound 'free' form of E2F1. Consistent with this, we find that the RB-associated histone deacetylase can deacetylate E2F1. These results identify acetylation as a novel regulatory modification that stimulates E2F1's activation functions. PMID- 10675336 TI - A single point mutation in TFIIA suppresses NC2 requirement in vivo. AB - Negative cofactor 2 (NC2) is a dimeric histone-fold complex that represses RNA polymerase II transcription through binding to TATA-box-binding protein (TBP) and inhibition of the general transcription factors TFIIA and TFIIB. Here we study molecular mechanisms of repression by human NC2 in vivo in yeast. Yeast NC2 genes are essential and can be exchanged with human NC2. The physiologically relevant regions of NC2 have been determined and shown to match the histone-fold dimerization motif. A suppressor screen based upon limiting concentrations of NC2beta yielded a cold-sensitive mutant in the yeast TFIIA subunit Toa1. The single point mutation in Toa1 alleviates the requirement for both subunits of NC2. Biochemical characterization indicated that mutant (mt)-Toa1 dimerizes well with Toa2; it supports specific recognition of the TATA box by TBP but forms less stable TBP-TFIIA-DNA complexes. Wild-type but not the mt-Toa1 can relieve NC2 effects in purified transcription systems. These data provide evidence for a dimeric NC2 complex that is in an equilibrium with TFIIA after the initial binding of TBP to promoter TATA boxes. PMID- 10675337 TI - The novel coactivator C1 (HCF) coordinates multiprotein enhancer formation and mediates transcription activation by GABP. AB - Transcription of the herpes simplex virus 1 (HSV-1) immediate early (IE) genes is determined by multiprotein enhancer complexes. The core enhancer assembly requires the interactions of the POU-homeodomain protein Oct-1, the viral transactivator alphaTIF and the cellular factor C1 (HCF). In this context, the C1 factor interacts with each protein to assemble the stable enhancer complex. In addition, the IE enhancer cores contain adjacent binding sites for other cellular transcription factors such as Sp1 and GA-binding protein (GABP). In this study, a direct interaction of the C1 factor with GABP is demonstrated, defining the C1 factor as the critical coordinator of the enhancer complex assembly. In addition, mutations that reduce the GABP transactivation potential also impair the C1-GABP interaction, indicating that the C1 factor functions as a novel coactivator of GABP-mediated transcription. The interaction and coordinated assembly of the enhancer proteins by the C1 factor may be critical for the regulation of the HSV lytic-latent cycle. PMID- 10675338 TI - Activation of orphan receptor-mediated transcription by Ca(2+)/calmodulin dependent protein kinase IV. AB - Retinoid-related receptor alpha (RORalpha) is an orphan nuclear receptor that constitutively activates transcription from its cognate response element. We show that RORalpha is Ca(2+ )responsive, and a Ca(2+)/calmodulin-independent form of Ca(2+)/calmodulin-dependent protein kinase IV (CaMKIV) potentiates RORalpha dependent transcription 20- to 30-fold. Other orphan receptors including RORalpha2, RORgamma and COUP-TFI are also potentiated by CaMKIV. Transcriptional activation by CaMKIV is orphan receptor selective and does not occur with either the thyroid hormone or estrogen receptor. CaMKIV does not phosphorylate RORalpha or its ligand-binding domain (LBD) in vitro, although the LBD is essential for transactivation. Therefore, the RORalpha LBD was used in the mammalian two-hybrid assay to identify a single class of small peptide molecules containing LXXLL motifs that interacted with greater affinity in the presence of CaMKIV. This class of peptides antagonized activation of orphan receptor-mediated transcription by CaMKIV. These studies demonstrate a pivotal role for CaMKIV in the regulation of orphan receptor-mediated transcription. PMID- 10675339 TI - Selective cell cycle transcription requires membrane synthesis in Caulobacter. AB - Caulobacter crescentus divides asymmetrically and creates distinct polar membrane surfaces that partition during the cell cycle to distinct cell progeny. Blocking membrane synthesis prevented transcription from selective promoters involved in asymmetric cell division. Transcription from sigma-54-dependent flagellar promoters was blocked completely; however, transcription from the CtrA response regulator-dependent flagellar promoters was activated but reduced. Transcription from the ccrM (DNA methylation) promoter and the che (chemosensory) promoter was also blocked completely. Transcription from a strong promoter at the chromosome replication origin was first stopped then induced by blocked membrane synthesis. We propose a feedback control coupling membrane synthesis to transcription that selectively supports membrane-associated processes such as flagellar assembly, chemosensory biogenesis and chromosome replication. PMID- 10675340 TI - Compartmentalization of transcription and translation in Bacillus subtilis. AB - Using fusions of green fluorescent protein to subunits of RNA polymerase (RNAP) and ribosomes, we have investigated the subcellular localization of the transcriptional and translational machinery in the bacterium Bacillus subtilis. Unexpectedly, we found that RNAP resides principally within the nucleoid. Conversely, ribosomes localized almost exclusively outside the nucleoid, concentrating particularly towards sites of cell division. This zonal localization was not dependent on cell division and is probably due, at least in part, to exclusion from the nucleoid. Dual labelling of RNAP and ribosomes was used to confirm the spatial separation of the two processes. We conclude that, even in the absence of a nuclear membrane, transcription and translation occur predominantly in separate functional domains. At higher growth rates, concentrations of RNAP developed, probably representing the sites of rRNA synthesis. These may represent a further spatial specialization, possibly equivalent to the eukaryotic nucleolus. PMID- 10675341 TI - On how a transcription factor can avoid its proteolytic activation in the absence of signal transduction. AB - In response to alkaline ambient pH, the Aspergillus nidulans PacC transcription factor mediating pH regulation of gene expression is activated by proteolytic removal of a negative-acting C-terminal domain. We demonstrate interactions involving the approximately 150 C-terminal PacC residues and two regions located immediately downstream of the DNA binding domain. Our data indicate two full length PacC conformations whose relative amounts depend upon ambient pH: one 'open' and accessible for processing, the other 'closed' and inaccessible. The location of essential determinants for proteolytic processing within the two more upstream interacting regions probably explains why the interactions prevent processing, whereas the direct involvement of the C-terminal region in processing preventing interactions explains why C-terminal truncating mutations result in alkalinity mimicry and pH-independent processing. A mutant PacC deficient in pH signal response and consequent processing behaves as though locked in the 'closed' form. Single-residue substitutions, obtained as mutations bypassing the need for pH signal transduction, identify crucial residues in each of the three interactive regions and overcome the processing deficiency in the 'permanently closed' mutant. PMID- 10675342 TI - Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation. AB - Hepatitis C virus (HCV) is the major etiological agent of blood-borne non-A non-B hepatitis and a leading cause of liver cirrhosis and hepatocellular carcinoma worldwide. HCV core protein is a multifunctional protein with regulatory functions in cellular transcription and virus-induced transformation and pathogenesis. Here we report on the identification of a bZIP nuclear transcription protein as an HCV core cofactor for transformation. This bZIP factor, designated LZIP, activates CRE-dependent transcription and regulates cell proliferation. Loss of LZIP function in NIH 3T3 cells triggers morphological transformation and anchorage-independent growth. We show that HCV core protein aberrantly sequesters LZIP in the cytoplasm, inactivates LZIP function and potentiates cellular transformation. Our findings suggest that LZIP might serve a novel cellular tumor suppressor function that is targeted by the HCV core. PMID- 10675343 TI - Hsp15: a ribosome-associated heat shock protein. AB - We are analyzing highly conserved heat shock genes of unknown or unclear function with the aim of determining their cellular role. Hsp15 has previously been shown to be an abundant nucleic acid-binding protein whose synthesis is induced massively at the RNA level upon temperature upshift. We have now identified that the in vivo target of Hsp15 action is the free 50S ribosomal subunit. Hsp15 binds with very high affinity (K(D) <5 nM) to this subunit, but only when 50S is free, not when it is part of the 70S ribosome. In addition, the binding of Hsp15 appears to correlate with a specific state of the mature, free 50S subunit, which contains bound nascent chain. This provides the first evidence for a so far unrecognized abortive event in translation. Hsp15 is suggested to be involved in the recycling of free 50S subunits that still carry a nascent chain. This gives Hsp15 a very different functional role from all other heat shock proteins and points to a new aspect of translation. PMID- 10675344 TI - Structure of Hsp15 reveals a novel RNA-binding motif. AB - We have solved the crystal structure of the heat shock protein Hsp15, a newly isolated and very highly inducible heat shock protein that binds the ribosome. Comparison of its structure with those of two RNA-binding proteins, ribosomal protein S4 and threonyl-tRNA synthetase, reveals a novel RNA-binding motif. This newly recognized motif is remarkably common, present in at least eight different protein families that bind RNA. The motif's surface is populated by conserved, charged residues that define a likely RNA-binding site. An intriguing pattern emerges: stress proteins, ribosomal proteins and tRNA synthetases repeatedly share a conserved motif. This may imply a hitherto unrecognized functional similarity between these three protein classes. PMID- 10675345 TI - DNA bending and a flip-out mechanism for base excision by the helix-hairpin-helix DNA glycosylase, Escherichia coli AlkA. AB - The Escherichia coli AlkA protein is a base excision repair glycosylase that removes a variety of alkylated bases from DNA. The 2.5 A crystal structure of AlkA complexed to DNA shows a large distortion in the bound DNA. The enzyme flips a 1-azaribose abasic nucleotide out of DNA and induces a 66 degrees bend in the DNA with a marked widening of the minor groove. The position of the 1-azaribose in the enzyme active site suggests an S(N)1-type mechanism for the glycosylase reaction, in which the essential catalytic Asp238 provides direct assistance for base removal. Catalytic selectivity might result from the enhanced stacking of positively charged, alkylated bases against the aromatic side chain of Trp272 in conjunction with the relative ease of cleaving the weakened glycosylic bond of these modified nucleotides. The structure of the AlkA-DNA complex offers the first glimpse of a helix-hairpin-helix (HhH) glycosylase complexed to DNA. Modeling studies suggest that other HhH glycosylases can bind to DNA in a similar manner. PMID- 10675346 TI - The numbers of individual mitochondrial DNA molecules and mitochondrial DNA nucleoids in yeast are co-regulated by the general amino acid control pathway. AB - Mitochondrial DNA (mtDNA) is inherited as a protein-DNA complex (the nucleoid). We show that activation of the general amino acid response pathway in rho(+) and rho(-) petite cells results in an increased number of nucleoids without an increase in mtDNA copy number. In rho(-) cells, activation of the general amino acid response pathway results in increased intramolecular recombination between tandemly repeated sequences of rho(-) mtDNA to produce small, circular oligomers that are packaged into individual nucleoids, resulting in an approximately 10 fold increase in nucleoid number. The parsing of mtDNA into nucleoids due to general amino acid control requires Ilv5p, a mitochondrial protein that also functions in branched chain amino acid biosynthesis, and one or more factors required for mtDNA recombination. Two additional proteins known to function in mtDNA recombination, Abf2p and Mgt1p, are also required for parsing mtDNA into a larger number of nucleoids, although expression of these proteins is not under general amino acid control. Increased nucleoid number leads to increased mtDNA transmission, suggesting a mechanism to enhance mtDNA inheritance under amino acid starvation conditions. PMID- 10675347 TI - Tn10 transpososome assembly involves a folded intermediate that must be unfolded for target capture and strand transfer. AB - Tn10 transposition, like all transposition reactions examined thus far, involves assembly of a stable protein-DNA transpososome, containing a pair of transposon ends, within which all chemical events occur. We report here that stable Tn10 pre cleavage transpososomes occur in two conformations: a folded form which contains the DNA-bending factor IHF and an unfolded form which lacks IHF. Functional analysis shows that both forms undergo double strand cleavage at the transposon ends but that only the unfolded form is competent for target capture (and thus for strand transfer to target DNA). Additional studies reveal that formation of any type of stable transpososome, folded or unfolded, requires not only IHF but also non-specific transposase-DNA contacts immediately internal to the IHF binding site, implying the occurrence of a topo- logically closed loop at the transposon end. Overall, transpososome assembly must proceed via a folded intermediate which, however, must be unfolded in order for intermolecular transposition to occur. These and other results support key features of a recently proposed model for transpososome assembly and morphogenesis. PMID- 10675348 TI - Epigenetics and its role in disease. PMID- 10675349 TI - Beckwith-Wiedemann syndrome: imprinting in clusters revisited. PMID- 10675350 TI - NF-kappaB determines localization and features of cell death in epidermis. AB - Specialized forms of physiologic cell death lacking certain characteristic morphologic features of apoptosis occur in terminally differentiating tissues, such as in the outer cell layers of epidermis. In these cell layers, NF-kappaB translocates from the cytoplasm to the nucleus and induces target gene expression. In light of its potent role in regulating apoptotic cell death in other tissues, NF-kappaB activation in these cells suggests that this transcription factor regulates cell death during terminal differentiation. Here, we show that NF-kappaB protects normal epithelial cells from apoptosis induced by both TNFalpha and Fas, whereas NF-kappaB blockade enhances susceptibility to death via both pathways. Expression of IkappaBalphaM under control of keratin promoter in transgenic mice caused a blockade of NF-kappaB function in the epidermis and provoked premature spontaneous cell death with apoptotic features. In normal tissue, expression of the known NF-kappaB-regulated antiapoptotic factors, TRAF1, TRAF2, c-IAP1, and c-IAP2, is most pronounced in outer epidermis. In transgenic mice, NF-kappaB blockade suppressed this expression, whereas NF kappaB activation augmented it, consistent with regulation of cell death by these NF-kappaB effector proteins. These data identify a new role for NF-kappaB in preventing premature apoptosis in cells committed to undergoing physiologic cell death and indicate that, in stratified epithelium, such cell death normally proceeds via a distinct pathway that is resistant to NF-kappaB and its antiapoptotic target effector genes. PMID- 10675351 TI - The basic helix-loop-helix transcription factor, dHAND, is required for vascular development. AB - Reciprocal interactions between vascular endothelial cells and vascular mesenchymal cells are essential for angiogenesis. Here we show that the basic helix-loop-helix transcription factor, dHAND/Hand2, is expressed in the developing vascular mesenchyme and its derivative, vascular smooth muscle cells (VSMCs). Targeted deletion of the dHAND gene in mice revealed severe defects of embryonic and yolk sac vascular development by embryonic day 9.5. Vascular endothelial cells expressed most markers of differentiation. Vascular mesenchymal cells migrated appropriately but failed to make contact with vascular endothelial cells and did not differentiate into VSMCs. In a screen for genes whose expression was dependent upon dHAND (using subtractive hybridization comparing wild-type and dHAND-null hearts), the VEGF(165) receptor, neuropilin-1, was found to be downregulated in dHAND mutants. These results suggest that dHAND is required for vascular development and regulates angiogenesis, possibly through a VEGF signaling pathway. PMID- 10675352 TI - Surgical implantation of adipose tissue reverses diabetes in lipoatrophic mice. AB - In lipoatrophic diabetes, a lack of fat is associated with insulin resistance and hyperglycemia. This is in striking contrast to the usual association of diabetes with obesity. To understand the underlying mechanisms, we transplanted adipose tissue into A-ZIP/F-1 mice, which have a severe form of lipoatrophic diabetes. Transplantation of wild-type fat reversed the hyperglycemia, dramatically lowered insulin levels, and improved muscle insulin sensitivity, demonstrating that the diabetes in A-ZIP/F-1 mice is caused by the lack of adipose tissue. All aspects of the A-ZIP/F-1 phenotype including hyperphagia, hepatic steatosis, and somatomegaly were either partially or completely reversed. However, the improvement in triglyceride and FFA levels was modest. Donor fat taken from parametrial and subcutaneous sites was equally effective in reversing the phenotype. The beneficial effects of transplantation were dose dependent and required near-physiological amounts of transplanted fat. Transplantation of genetically modified fat into A-ZIP/F-1 mice is a new and powerful technique for studying adipose physiology and the metabolic and endocrine communication between adipose tissue and the rest of the body. PMID- 10675353 TI - Multidrug resistance protein 1 protects the choroid plexus epithelium and contributes to the blood-cerebrospinal fluid barrier. AB - Multidrug resistance protein 1 (MRP1) is a transporter protein that helps to protect normal cells and tumor cells against the influx of certain xenobiotics. We previously showed that Mrp1 protects against cytotoxic drugs at the testis blood barrier, the oral epithelium, and the kidney urinary collecting duct tubules. Here, we generated Mrp1/Mdr1a/Mdr1b triple-knockout (TKO) mice, and used them together with Mdr1a/Mdr1b double-knockout (DKO) mice to study the contribution of Mrp1 to the tissue distribution and pharmacokinetics of etoposide. We observed increased toxicity in the TKO mice, which accumulated etoposide in brown adipose tissue, colon, salivary gland, heart, and the female urogenital system. Immunohistochemical staining revealed the presence of Mrp1 in the oviduct, uterus, salivary gland, and choroid plexus (CP) epithelium. To explore the transport function of Mrp1 in the CP epithelium, we used TKO and DKO mice cannulated for cerebrospinal fluid (CSF). We show here that the lack of Mrp1 protein causes etoposide levels to increase about 10-fold in the CSF after intravenous administration of the drug. Our results indicate that Mrp1 helps to limit tissue distribution of certain drugs and contributes to the blood-CSF drug permeability barrier. PMID- 10675354 TI - Improved insulin-sensitivity in mice heterozygous for PPAR-gamma deficiency. AB - The thiazolidinedione class of insulin-sensitizing, antidiabetic drugs interacts with peroxisome proliferator-activated receptor gamma (PPAR-gamma). To gain insight into the role of this nuclear receptor in insulin resistance and diabetes, we conducted metabolic studies in the PPAR-gamma gene knockout mouse model. Because homozygous PPAR-gamma-null mice die in development, we studied glucose metabolism in mice heterozygous for the mutation (PPAR-gamma(+/-) mice). We identified no statistically significant differences in body weight, basal glucose, insulin, or FFA levels between the wild-type (WT) and PPAR-gamma(+/-) groups. Nor was there a difference in glucose excursion between the groups of mice during oral glucose tolerance test, but insulin concentrations of the WT group were greater than those of the PPAR-gamma(+/-) group, and insulin-induced increase in glucose disposal rate was significantly increased in PPAR-gamma(+/-) mice. Likewise, the insulin-induced suppression of hepatic glucose production was significantly greater in the PPAR-gamma(+/-) mice than in the WT mice. Taken together, these results indicate that - counterintuitively - although pharmacological activation of PPAR-gamma improves insulin sensitivity, a similar effect is obtained by genetically reducing the expression levels of the receptor. PMID- 10675355 TI - Decreased neointimal formation in Mac-1(-/-) mice reveals a role for inflammation in vascular repair after angioplasty. AB - Inflammation plays an essential role in the initiation and progression of atherosclerosis, but its role in vascular repair after mechanical arterial injury (i.e., percutaneous transluminal coronary angioplasty, PTCA) is unknown. In animal models of vascular injury, leukocytes are recruited as a precursor to intimal thickening. Furthermore, markers of leukocyte activation - in particular, increased expression of the beta2-integrin Mac-1 (alphaMbeta2, or CD11b/CD18), which is responsible for firm leukocyte adhesion to platelets and fibrinogen on denuded vessels - predict restenosis after PTCA. To determine whether Mac-1 mediated leukocyte recruitment is causally related to neointimal formation, we subjected mice lacking Mac-1 to a novel form of mechanical carotid artery dilation and complete endothelial denudation. We now report that the selective absence of Mac-1 impairs transplatelet leukocyte migration into the vessel wall, reducing leukocyte accumulation over time. Diminished medial leukocyte accumulation was accompanied by markedly reduced neointimal thickening after vascular injury. These data establish a role for inflammation in neointimal thickening and suggest that leukocyte recruitment to mechanically injured arteries may prevent restenosis. PMID- 10675356 TI - Differential immune responses to alpha-gal epitopes on xenografts and allografts: implications for accommodation in xenotransplantation. AB - Xenograft recipients produce large amounts of high-affinity anti-Gal IgG in response to Galalpha1-3Galbeta1- 4GlcNAc-R (alpha-gal) epitopes on the graft. In contrast, ABO-mismatched allograft recipients undergo "accommodation," a state of very weak immune response to ABO antigens. These differences in anti-carbohydrate immune response were studied in alpha1,3galactosyltransferase knock-out mice. Pig kidney membranes administered to these mice elicited extensive production of anti Gal IgG, whereas allogeneic kidney membranes expressing alpha-gal epitopes elicited only a weak anti-Gal IgM response. Anti-Gal IgG response to xenograft membranes depended on helper T cell activation and was inhibited by anti-CD40L antibody. These T cells were activated by xenopeptides and not by alpha-gal epitopes. Moreover, allogeneic cell membranes manipulated to express xenoproteins also induced anti-Gal IgG response. Xenoglycoproteins with alpha-gal epitopes are processed by anti-Gal B cells. Xenopeptides presented by these cells activate a large repertoire of helper T cells required for the differentiation of anti-Gal B cells into cells secreting anti-Gal IgG. Alloglycoproteins with alpha- gal epitopes have very few immunogenic peptides and fail to activate helper T cells. Similarly, ineffective helper T-cell activation prevents a strong immune response to blood group antigens in ABO-mismatched allograft recipients, thus enabling the development of accommodation. PMID- 10675357 TI - Insulin resistance differentially affects the PI 3-kinase- and MAP kinase mediated signaling in human muscle. AB - The broad nature of insulin resistant glucose metabolism in skeletal muscle of patients with type 2 diabetes suggests a defect in the proximal part of the insulin signaling network. We sought to identify the pathways compromised in insulin resistance and to test the effect of moderate exercise on whole-body and cellular insulin action. We conducted euglycemic clamps and muscle biopsies on type 2 diabetic patients, obese nondiabetics and lean controls, with and without a single bout of exercise. Insulin stimulation of the phosphatidylinositol 3 kinase (PI 3-kinase) pathway, as measured by phosphorylation of the insulin receptor and IRS-1 and by IRS protein association with p85 and with PI 3-kinase, was dramatically reduced in obese nondiabetics and virtually absent in type 2 diabetic patients. Insulin stimulation of the MAP kinase pathway was normal in obese and diabetic subjects. Insulin stimulation of glucose-disposal correlated with association of p85 with IRS-1. Exercise 24 hours before the euglycemic clamp increased phosphorylation of insulin receptor and IRS-1 in obese and diabetic subjects but did not increase glucose uptake or PI 3-kinase association with IRS 1 upon insulin stimulation. Thus, insulin resistance differentially affects the PI 3-kinase and MAP kinase signaling pathways, and insulin-stimulated IRS-1 association with PI 3-kinase defines a key step in insulin resistance. PMID- 10675358 TI - A novel mutation in DAX1 causes delayed-onset adrenal insufficiency and incomplete hypogonadotropic hypogonadism. AB - Mutations in the DAX1 gene cause X-linked adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HHG). In affected boys, primary adrenal insufficiency occurs soon after birth or during early childhood; HHG is recognized at the expected time of puberty. In this report, we describe the novel phenotype of a man who presented with apparently isolated adrenal insufficiency at 28 years of age. Examination revealed partial pubertal development and undiagnosed incomplete HHG. Gonadotropin therapy did not improve his marked oligospermia, suggesting a concomitant primary testicular abnormality. Genomic analysis revealed a novel missense mutation, I439S, in DAX1. The mutant DAX-1 protein was studied for its ability to function as a transcriptional repressor of target genes. Consistent with the patient's mild clinical phenotype, the I439S mutation conferred intermediate levels of repressor activity of DAX-1 when compared with mutations associated with classic AHC. This unique case extends the clinical spectrum of AHC to include delayed-onset primary adrenal insufficiency in adulthood and milder forms of HHG. Furthermore, in accordance with findings in Ahch (Dax1) knockout mice, the clinical features in this patient suggest that DAX 1 function is required for spermatogenesis in humans, independent of its known effects on gonadotropin production. PMID- 10675359 TI - Colchicine protects mice from the lethal effect of an agonistic anti-Fas antibody. AB - The aim of this study was to determine whether colchicine, which has been reported to protect against various hepatotoxic insults, influences the susceptibility of mice to the agonistic anti-Fas antibody, Jo2. All mice that were pretreated with colchicine (2 mg/kg) survived the lethal challenge of intraperitoneal administration of 10 microg of Jo2, whereas all control mice pretreated with gamma-lumicolchicine succumbed to the challenge. Twelve micrograms of Jo2 killed less than half of colchicine-pretreated mice and its lethal effects were delayed relative to control mice, which all died within 8 hours. Other microtubule-disrupting agents such as Taxol, vinblastine, and nocodazole also improved the survival of mice treated with the lethal dose of Jo2. Histologic examination showed that colchicine protected against Jo2-induced fulminant liver injury, and TUNEL assay demonstrated that colchicine protected against massive apoptosis of hepatocytes. Hepatocytes isolated from colchicine pretreated mice exhibited decreased susceptibility to Jo2-induced apoptosis. In addition, colchicine pretreatment reduced surface expression of Fas and decreased Jo2- and TNF-alpha-induced apoptosis of cultured hepatocytes in the presence of actinomycin D, but did not affect the susceptibility of cultured sinusoidal endothelial cells to Jo2-induced apoptosis. Remarkably, Fas and TNF receptor-1 mRNA and intracellular protein levels increased after colchicine treatment, indicating that colchicine protects against death ligand-induced apoptosis in the liver by decreasing death-receptor targeting to the cell surface. PMID- 10675360 TI - Heparin-binding EGF-like growth factor contributes to reduced glomerular filtration rate during glomerulonephritis in rats. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a member of the epidermal growth factor (EGF) family, is expressed during inflammatory and pathological conditions. We have cloned the rat HB-EGF and followed the expression of HB-EGF in rat kidneys treated with anti- glomerular basement membrane (anti-GBM) antibody (Ab) to induce glomerulonephritis (GN). We observed glomerular HB-EGF mRNA and protein within 30 minutes of Ab administration and showed by in situ hybridization that glomerular HB-EGF mRNA expression was predominantly in mesangial and epithelial cells. Expression of HB-EGF correlated with the onset of decreased renal function in this model. To test the direct effect of HB-EGF on renal function, we infused the renal cortex with active rHB EGF, prepared from transfected Drosophila melanogaster cells. This treatment induced a significant decrease in single nephron GFR (SNGFR), single nephron plasma flow, and glomerular ultrafiltration coefficient and an increase in the glomerular capillary hydrostatic pressure gradient. In addition, anti-HB-EGF Ab administered just before anti-GBM Ab blocked the fall in SNGFR and GFR at 90 minutes without any change in the glomerular histologic response. These studies suggest that HB-EGF expressed early in the anti-GBM Ab GN model contributes to the observed acute glomerular hemodynamic alterations. PMID- 10675361 TI - Diazepam-binding inhibitor mediates feedback regulation of pancreatic secretion and postprandial release of cholecystokinin. AB - Recently, we isolated a trypsin-sensitive cholecystokinin-releasing peptide (CCK RP) from porcine and rat intestinal mucosa. The amino acid sequence of this peptide was determined to be identical to that of the diazepam-binding inhibitor (DBI). To test the role of DBI in pancreatic secretion and responses to feeding, we used pancreaticobiliary and intestinal cannula to divert bile-pancreatic juice from anesthetized rats. Within 2 hours, this treatment caused a 2-fold increase in pancreatic protein output and a >10-fold increase in plasma CCK. Luminal DBI levels increased 4-fold. At 5 hours after diversion of bile-pancreatic juice, each of these measures returned to basal levels. Intraduodenal infusion of peptone evoked a 5-fold increase in the concentration of luminal DBI. In separate studies, we demonstrated that intraduodenal administration of antiserum to a DBI peptide specifically abolished pancreatic secretion and the increase in plasma CCK levels after diversion of bile-pancreatic juice. To demonstrate that DBI mediates the postprandial rise in plasma CCK levels, we showed that intraduodenal administration of 5% peptone induced dramatic increases in pancreatic secretion and plasma CCK, effects that could be blocked by intraduodenal administration of anti-DBI antiserum. Hence, DBI, a trypsin-sensitive CCK-RP secreted from the proximal small bowel, mediates the feedback regulation of pancreatic secretion and the postprandial release of CCK. PMID- 10675362 TI - Adenosine and inosine increase cutaneous vasopermeability by activating A(3) receptors on mast cells. AB - Adenosine has potent effects on both the cardiovascular and immune systems. Exposure of tissues to adenosine results in increased vascular permeability and extravasation of serum proteins. The mechanism by which adenosine brings about these physiological changes is poorly defined. Using mice deficient in the A(3) adenosine receptor (A(3)AR), we show that increases in cutaneous vascular permeability observed after treatment with adenosine or its principal metabolite inosine are mediated through the A(3)AR. Adenosine fails to increase vascular permeability in mast cell-deficient mice, suggesting that this tissue response to adenosine is mast cell-dependent. Furthermore, this response is independent of activation of the high-affinity IgE receptor (FcepsilonR1) by antigen, as adenosine is equally effective in mediating these changes in FcepsilonR1 beta chain-deficient mice. Together these results support a model in which adenosine and inosine induce changes in vascular permeability indirectly by activating mast cells, which in turn release vasoactive substances. The demonstration in vivo that adenosine, acting through a specific receptor, can provoke degranulation of this important tissue-based effector cell, independent of antigen activation of the high-affinity IgE receptor, supports an important role for this nucleoside in modifying the inflammatory response. PMID- 10675363 TI - C-reactive protein binding to FcgammaRIIa on human monocytes and neutrophils is allele-specific. AB - C-reactive protein (CRP) is involved in host defense, regulation of inflammation, and modulation of autoimmune disease. Although the presence of receptors for CRP on phagocytes has been inferred for years, their identity was determined only recently. FcgammaRIa, the high-affinity IgG receptor, binds CRP with low affinity, whereas FcgammaRIIa, the low-affinity IgG receptor, binds CRP with high affinity. Because the single nucleotide polymorphism in FcgammaRIIA - which encodes histidine or arginine at position 131 - strongly influences IgG2 binding, we determined this polymorphism's effect on CRP binding. CRP bound with high avidity to monocytes and neutrophils from FcgammaRIIA R-131 homozygotes, and binding was inhibited by the R-specific mAb 41H16. CRP showed decreased binding to cells from FcgammaRIIA H-131 homozygotes (which bind IgG2 with high affinity). However, IFN-gamma enhanced FcgammaRI expression by H-131 monocytes and increased CRP binding. FcgammaRIIa heterozygotes showed intermediate binding. CRP initiated increases in [Ca(2+)](i) in PMN from R-131, but not from H-131 homozygotes. These data provide direct genetic evidence for FcgammaRIIa as the functional, high affinity CRP receptor on leukocytes while emphasizing the reciprocal relationship between IgG and CRP binding avidities. This counterbalance may affect the contribution of FcgammaRIIA alleles to host defense and autoimmunity. PMID- 10675364 TI - Syntaxin 1A is expressed in airway epithelial cells, where it modulates CFTR Cl( ) currents. AB - The CFTR Cl(-) channel controls salt and water transport across epithelial tissues. Previously, we showed that CFTR-mediated Cl(-) currents in the Xenopus oocyte expression system are inhibited by syntaxin 1A, a component of the membrane trafficking machinery. This negative modulation of CFTR function can be reversed by soluble syntaxin 1A peptides and by the syntaxin 1A binding protein, Munc-18. In the present study, we determined whether syntaxin 1A is expressed in native epithelial tissues that normally express CFTR and whether it modulates CFTR currents in these tissues. Using immunoblotting and immunofluorescence, we observed syntaxin 1A in native gut and airway epithelial tissues and showed that epithelial cells from these tissues express syntaxin 1A at >10-fold molar excess over CFTR. Syntaxin 1A is seen near the apical cell surfaces of human bronchial airway epithelium. Reagents that disrupt the CFTR-syntaxin 1A interaction, including soluble syntaxin 1A cytosolic domain and recombinant Munc-18, augmented cAMP-dependent CFTR Cl(-) currents by more than 2- to 4-fold in mouse tracheal epithelial cells and cells derived from human nasal polyps, but these reagents did not affect CaMK II-activated Cl(-) currents in these cells. PMID- 10675366 TI - MEMORANDUM FOR: science writers and editors on the journal press list : genetic similarity found between ductal carcinomas In situ and their recurrences PMID- 10675365 TI - RXRalpha overexpression in cardiomyocytes causes dilated cardiomyopathy but fails to rescue myocardial hypoplasia in RXRalpha-null fetuses. AB - Retinoid X receptor alpha-null (RXRalpha-null) mutants exhibit hypoplasia of their ventricular myocardium and die at the fetal stage. In the present study, we wished to determine whether transgenic re-expression of RXRalpha in mutant cardiac myocytes could rescue these defects. Two transgenic mouse lines specifically overexpressing an RXRalpha protein in cardiomyocytes were generated, using the cardiac alpha-myosin heavy chain (alpha-MHC) promoter. Breeding the high copy number transgenic line onto an RXRalpha-null genetic background did not prevent the myocardial hypoplasia and fetal lethality associated with the RXRalpha(-/-) genotype, even though the transgene was expressed in the ventricles as early as 10. 5 days post-coitum. These data suggest that the RXRalpha function involved in myocardial growth may correspond to a non-cell-autonomous requirement forsignal orchestrating the growth and differentiation of myocytes. Interestingly, the adult transgenic mice developed a dilated cardiomyopathy, associated with myofibrillar abnormalities and specific deficiencies in respiratory chain complexes I and II, thus providing an additional model for this genetically complex disease. PMID- 10675367 TI - Relation of a recurrent intraductal carcinoma (ductal carcinoma in situ) to the primary tumor. PMID- 10675368 TI - Is ATP (adenosine 5'-triphosphate), like STP, a performance-enhancing additive for the tanks of cancer patients? PMID- 10675371 TI - HER2 testing methods PMID- 10675370 TI - Common techniques used to detect HER2 PMID- 10675369 TI - Experts debate value of HER2 testing methods. PMID- 10675372 TI - San Antonio Breast Cancer Symposium explores DCIS "battleground". PMID- 10675373 TI - Stat bite: Use of fecal occult blood test in the United States. PMID- 10675374 TI - Arthritis drug approved for polyp prevention blazes trail for other prevention trials. PMID- 10675375 TI - Old drugs find new life PMID- 10675376 TI - NIH receives a 14.9% increase in fiscal year 2000. PMID- 10675377 TI - Celecoxib trials under Way PMID- 10675378 TI - NJ insurers agree to pay for trials. PMID- 10675379 TI - History of breast-feeding in relation to breast cancer risk: a review of the epidemiologic literature. AB - The purpose of this review is to critically evaluate the collective epidemiologic evidence that a history of breast-feeding may decrease the risk of breast cancer. Original data for inclusion were identified through a MEDLINE(R) search of the English language literature from 1966 through 1998. To date, virtually all epidemiologic data regarding breast-feeding and breast cancer risk are derived from case-control studies, which vary according to classification of breast feeding history. Overall, the evidence with respect to "ever" breast-feeding remains inconclusive, with results indicating either no association or a rather weak protective effect against breast cancer. An inverse association between increasing cumulative duration of breast-feeding and breast cancer risk among parous women has been reported in some, but not all, studies; the failure to detect an association in some Western populations may be due to the low prevalence of prolonged breast-feeding. It appears that the protective effect, if any, of long-term breast-feeding is stronger among, or confined to, premenopausal women. It has been hypothesized that an apparently protective effect of breast feeding may be due to elevated breast cancer risk among women who discontinue breast-feeding or who take medication to suppress lactation; however, the evidence is limited and should be interpreted with caution. The biology underlying a protective effect of breast-feeding and why this should be restricted to premenopausal women remain unknown, although several mechanisms have been postulated (hormonal changes, such as reduced estrogen; removal of estrogens through breast fluid; excretion of carcinogens from breast tissue through breast-feeding; physical changes in the mammary epithelial cells, reflecting maximal differentiation; and delay of the re-establishment of ovulation). While breast-feeding is a potentially modifiable behavior, the practical implication of reduced breast cancer risk among premenopausal women with prolonged durations of breast-feeding may be of marginal importance, particularly in Western societies. PMID- 10675380 TI - Chromosomal alterations in ductal carcinomas in situ and their in situ recurrences. AB - BACKGROUND: Ductal carcinoma in situ (DCIS) recurs in the same breast following breast-conserving surgery in 5%-25% of patients, with the rate influenced by the presence or absence of involved surgical margins, tumor size and nuclear grade, and whether or not radiation therapy was performed. A recurrent lesion arising soon after excision of an initial DCIS may reflect residual disease, whereas in situ tumors arising after longer periods are sometimes considered to be second independent events. The purpose of this study was to determine the clonal relationship between initial DCIS lesions and their recurrences. METHODS: Comparative genomic hybridization (CGH) was used to compare chromosomal alterations in 18 initial DCIS lesions (presenting in the absence of invasive disease) and in their subsequent ipsilateral DCIS recurrences (detected from 16 months to 9.3 years later). RESULTS: Of the 18 tumor pairs, 17 showed a high concordance in their chromosomal alterations (median = 81%; range = 65%-100%), while one case showed no agreement between the paired samples (having two and 20 alterations, respectively). Morphologic characterization of the DCIS pairs showed clear similarities. The mean number of CGH changes was greater in the recurrent tumors than in the initial lesions (10.7 versus 8.8; P =.019). The most common changes in both the initial and the recurrent in situ lesions were gains involving chromosome 17q and losses involving chromosomes 8p and 17p. The degree of concordance was independent of the time interval before recurrence and of the presence of positive surgical margins. CONCLUSIONS: In this study, DCIS recurrences were clonally related to their primary lesions in most cases. This finding is consistent with treatment paradigms requiring wide surgical margins and/or postoperative radiation therapy. PMID- 10675381 TI - Randomized clinical trial of adenosine 5'-triphosphate in patients with advanced non-small-cell lung cancer. AB - BACKGROUND: Extracellular adenosine 5'-triphosphate (ATP) is involved in the regulation of a variety of biologic processes, including neurotransmission, muscle contraction, and liver glucose metabolism, via purinergic receptors. In nonrandomized studies involving patients with different tumor types including non small-cell lung cancer (NSCLC), ATP infusion appeared to inhibit loss of weight and deterioration of quality of life (QOL) and performance status. We conducted a randomized clinical trial to evaluate the effects of ATP in patients with advanced NSCLC (stage IIIB or IV). METHODS: Fifty-eight patients were randomly assigned to receive either 10 intravenous 30-hour ATP infusions, with the infusions given at 2- to 4-week intervals, or no ATP. Outcome parameters were assessed every 4 weeks until 28 weeks. Between-group differences were tested for statistical significance by use of repeated-measures analysis, and reported P values are two-sided. RESULTS: Twenty-eight patients were allocated to receive ATP treatment and 30 received no ATP. Mean weight changes per 4-week period were 1.0 kg (95% confidence interval [CI] = -1.5 to -0.5) in the control group and 0.2 kg (95% CI = -0.2 to +0.6) in the ATP group (P =.002). Serum albumin concentration declined by -1.2 g/L (95% CI= -2.0 to -0.4) per 4 weeks in the control group but remained stable (0.0 g/L; 95% CI = -0.3 to +0.3) in the ATP group (P =.006). Elbow flexor muscle strength declined by -5.5% (95% CI = -9.6% to -1. 4%) per 4 weeks in the control group but remained stable (0.0%; 95% CI= 1.4% to +1.4%) in the ATP group (P =.01). A similar pattern was observed for knee extensor muscles (P =.02). The effects of ATP on body weight, muscle strength, and albumin concentration were especially marked in cachectic patients (P =.0002, P =.0001, and P =. 0001, respectively, for ATP versus no ATP). QOL score changes per 4-week period in the ATP group showed overall less deterioration than in the control group-physical scores (-0.2% versus -2.4%; P =. 0002); functional scores (+0.4% versus -5.5%; P =.02); psychologic scores (-0.7% versus -2.4%; P =.11); overall QOL score (+0.1% versus -3.5%; P =.0001). CONCLUSIONS: This randomized trial demonstrates that ATP has beneficial effects on weight, muscle strength, and QOL in patients with advanced NSCLC. PMID- 10675382 TI - Effect of hormone replacement therapy on breast cancer risk: estrogen versus estrogen plus progestin. AB - BACKGROUND: Hormone replacement therapy (HRT) given as unopposed estrogen replacement therapy (ERT) gained widespread popularity in the United States in the 1960s and 1970s. Recent prescribing practices have favored combination HRT (CHRT), i.e., adding a progestin to estrogen for the entire monthly cycle (continuous combined replacement therapy [CCRT]) or a part of the cycle (sequential estrogen plus progestin therapy [SEPRT]). Few data exist on the association between CHRT and breast cancer risk. We determined the effects of CHRT on a woman's risk of developing breast cancer in a population-based, case control study. METHODS: Case subjects included those with incident breast cancers diagnosed over 4(1/2) years in Los Angeles County, CA, in the late 1980s and 1990s. Control subjects were neighborhood residents who were individually matched to case subjects on age and race. Case subjects and control subjects were interviewed in person to collect information on known breast cancer risk factors as well as on HRT use. Information on 1897 postmenopausal case subjects and on 1637 postmenopausal control subjects aged 55-72 years who had not undergone a simple hysterectomy was analyzed. Breast cancer risks associated with the various types of HRT were estimated as odds ratios (ORs) after adjusting simultaneously for the different forms of HRT and for known risk factors of breast cancer. All P values are two-sided. RESULTS: HRT was associated with a 10% higher breast cancer risk for each 5 years of use (OR(5) = 1.10; 95% confidence interval [CI] = 1.02 1.18). Risk was substantially higher for CHRT use (OR(5) = 1.24; 95% CI = 1.07 1.45) than for ERT use (OR(5) = 1. 06; 95% CI = 0.97-1.15). Risk estimates were higher for SEPRT (OR(5) = 1.38; 95% CI = 1.13-1.68) than for CCRT (OR(5) = 1.09; 95% CI = 0. 88-1.35), but this difference was not statistically significant. CONCLUSIONS: This study provides strong evidence that the addition of a progestin to HRT enhances markedly the risk of breast cancer relative to estrogen use alone. These findings have important implications for the risk-benefit equation for HRT in women using CHRT. PMID- 10675383 TI - Cigar smoking in men and risk of death from tobacco-related cancers. AB - BACKGROUND: Cigar consumption in the United States has increased dramatically since 1993, yet there are limited prospective data on the risk of cancer associated with cigar smoking. We examined the association between cigar smoking and death from tobacco-related cancers in a large, prospective cohort of U. S. men. METHODS: We used Cox proportional hazards models to analyze the relationship between cigar smoking at baseline in 1982 and mortality from cancers of the lung, oral cavity/pharynx, larynx, esophagus, bladder, and pancreas over 12 years of follow-up of the American Cancer Society's Cancer Prevention Study II cohort. A total of 137 243 men were included in the final analysis. Women were not included because we had no data on their cigar use. We excluded men who ever smoked cigarettes or pipes and adjusted all rate ratio (RR) estimates for age, alcohol use, and use of snuff or chewing tobacco. RESULTS: Current cigar smoking at baseline, as compared with never smoking, was associated with an increased risk of death from cancers of the lung (RR = 5.1; 95% confidence interval [CI] = 4.0 6.6), oral cavity/pharynx (RR = 4.0 [95% CI = 1.5-10.3]), larynx (RR = 10.3 [95% CI = 2.6-41.0]), and esophagus (RR = 1.8; 95% CI = 0.9-3.7). Although current cigar smokers overall did not appear to be at an increased risk of death from cancer of the pancreas (RR = 1.3; 95% CI = 0.9-1.9) or bladder (RR = 1.0; 95% CI = 0.4-2.3), there was an increased risk for current cigar smokers who reported that they inhaled the smoke (for pancreas, RR = 2.7; 95% CI = 1.5-4.8; for bladder, RR = 3.6; 95% CI = 1.3-9.9). CONCLUSIONS: Results from this large prospective study support a strong association between cigar smoking and mortality from several types of cancer. PMID- 10675384 TI - Restored expression of fragile histidine triad protein and tumorigenicity of cervical carcinoma cells. AB - BACKGROUND: Allelic losses in the short arm of chromosome 3 are common in cervical carcinomas. The fragile histidine triad (FHIT) gene at chromosome region 3p14.2 is a candidate tumor suppressor gene that may play a role in cervical tumorigenesis. We and others have identified aberrant FHIT transcripts and frequent loss of Fhit protein expression in primary cervical cancers and high grade noninvasive lesions but not in normal cervical tissues. The altered expression of FHIT may be due to somatic mutations or integration of human papillomavirus DNA at the FHIT locus. The purpose of this study was to determine whether ectopic expression of Fhit can suppress the tumorigenic properties of cervical cancer cells. METHODS: We employed infection with recombinant retroviruses as well as transfection of plasmid DNA to restore Fhit protein expression in cervical cancer cell lines lacking full-length FHIT transcripts and endogenous Fhit protein. The effects of Fhit expression on tumor cell morphology, anchorage-independent growth, and tumorigenicity in nude mice were examined. RESULTS: Stable overexpression of Fhit had no discernible effect on the tumorigenic properties of two cervical carcinoma cell lines or on a lung carcinoma cell line previously reported by others to be suppressed for tumorigenicity by Fhit. CONCLUSIONS: Restoration of Fhit expression does not suppress anchorage-independent growth or tumorigenicity of cervical carcinoma cell lines. However, it remains possible that FHIT inactivation may be important early in cervical tumor progression or that FHIT may suppress tumorigenesis in ways distinct from those measured by the assays employed in this study. PMID- 10675386 TI - RESPONSE: re: antiangiogenic activity of prostate-specific antigen PMID- 10675385 TI - Re: Antiangiogenic activity of prostate-specific antigen. PMID- 10675387 TI - Re: Enhancement of tumor response to gamma-radiation by an inhibitor of cyclooxygenase-2 enzyme. PMID- 10675388 TI - RESPONSE: re: enhancement of tumor response to gamma-radiation by an inhibitor of cyclooxygenase-2 enzyme PMID- 10675389 TI - Re: Dormancy of mammary carcinoma after mastectomy. PMID- 10675390 TI - RESPONSE: re: dormancy of mammary carcinoma after mastectomy PMID- 10675392 TI - PSYCHOSOMATICS: Information for Contributors. PMID- 10675391 TI - Consultation-Liaison Psychiatry: Drug-Drug Interactions Update. PMID- 10675393 TI - Identification of a pathogenicity determinant of Plum pox virus in the sequence encoding the C-terminal region of protein P3+6K(1). AB - A full-length genomic cDNA clone of a plum pox potyvirus (PPV) isolate belonging to the M strain (PPV-PS) has been cloned downstream from a bacteriophage T7 polymerase promoter and sequenced. Transcripts from the resulting plasmid, pGPPVPS, were infectious and, in herbaceous hosts, produced symptoms that differed from those of virus progeny of pGPPV, a full-length genomic cDNA clone of the D strain PPV-R. Viable PPV-R/-PS chimeric viruses were constructed by recombination of the cDNA clones in vitro. Analysis of plants infected with the different chimeras indicated that sequences encoding the most variable regions of the potyvirus genome, the P1 and capsid protein coding sequences, were not responsible for symptom differences between the two PPV isolates in herbaceous hosts. On the contrary, complex symptomatology determinants seem to be located in the central region of the PPV genome. The results indicate that a genomic fragment that encodes 173 aa from the C-terminal part of the P3+6K(1) coding region is enough to confer, on a PPV-R background, a PS phenotype in Nicotiana clevelandii. This pathogenicity determinant also participates in symptom induction in Pisum sativum, although the region defining the PS phenotype in this host is probably restricted to 74 aa. PMID- 10675394 TI - Mutations in the coat protein gene of plum pox virus suppress particle assembly, heterologous encapsidation and complementation in transgenic plants of Nicotiana benthamiana. AB - Two different motifs in the coat protein (CP) of Plum pox virus (PPV) (R(3015)Q(3016), D(3059)) were mutated by replacing the respective amino acids with others possessing different chemical properties. The mutated CP genes were introduced into an infectious full-length clone of PPV (p35PPV-NAT) to investigate their influence on systemic infection of transgenic wild-type PPV CP expressing and non-transgenic plants of Nicotiana benthamiana. All mutants failed to establish systemic infections in non-transgenic N. benthamiana plants, but were complemented by intact CP in transgenic plants. Moreover, the CP-RQ-D mutant (carrying mutations in both the RQ and D motifs) was introduced into p35PPV-NAT engineered to express beta-glucuronidase (GUS) for direct observation of systemic movement and particle assembly in N. benthamiana leaves. GUS-staining revealed that the CP mutant (RQ-D) was restricted to initially infected cells without forming virions. Systemic movement and particle assembly were restored in CP transgenic N. benthamiana plants. Finally, transgenic N. benthamiana plants were generated that expressed each of the three mutated CP genes. Homozygous T(2) lines were selected and tested for resistance to PPV. Immunogold labelling and electron microscopy revealed that heterologous encapsidation with challenging Chilli veinal mottle virus and Potato virus Y was suppressed in these lines. In addition, assembly mutants did not complement CP-defective p35PPV-NAT. The possible use of modified viral CP genes for the production of virus-resistant transgenic plants, thereby reducing the putative risks of heterologous encapsidation and complementation, is discussed. PMID- 10675395 TI - Heterogeneity in the 3'-terminal untranslated region of tobacco mild green mosaic tobamoviruses from Nicotiana glauca resulting in variants with three or six pseudoknots. AB - Isolates of tobacco mild green mosaic tobamovirus (TMGMV) were obtained from 58 plants of Nicotiana glauca in southern California and placed in one of two groups (Small type and Large type) based on the size of the subgenomic RNA for the coat protein (CP). The CP sequence differed by no more than one amino acid for the two types, and the Small type was identical to that published for TMGMV. Thirty-six of the isolates had a double-stranded (ds)RNA profile that matched that of type TMGMV, and the nucleotide sequence of the 3' untranslated region (3'UTR) of six of these isolates was similar to the published sequence of TMGMV. Twenty-two isolates had a larger dsRNA for the CP subgenomic RNA. Six of these were sequenced and all had a repeat sequence of between 147 and 165 bases in the part of the 3'UTR that is involved in the formation of pseudoknots. These novel but common isolates are predicted to have six rather than three pseudoknots. Small types (three pseudoknots=type TMGMV) yielded twice as much virus after purification as Large types (six pseudoknots). The two groups of isolates could be distinguished in N. rustica (Large type, but not Small type gave a systemic infection), and N. clevelandii (Small type but not Large type induced systemic lethal necrosis). Almost all isolates of TMGMV used in this study were initially associated with satellite tobacco mosaic virus (STMV), and both types supported STMV experimentally. PMID- 10675396 TI - Transgene translatability increases effectiveness of replicase-mediated resistance to cucumber mosaic virus. AB - Transgenic tobacco plants expressing an altered form of the 2a replicase gene from the Fny strain of Cucumber mosaic virus (CMV) exhibit suppressed virus replication and restricted virus movement when inoculated mechanically or by aphid vectors. Additional transformants have been generated which contain replicase gene constructs designed to determine the role(s) of transgene mRNA and/or protein in resistance. Resistance to systemic disease caused by CMV, as well as delayed infection, was observed in several lines of transgenic plants which were capable of expressing either full-length or truncated replicase proteins. In contrast, among plants which contained nontranslatable transgene constructs, only one of 61 lines examined exhibited delays or resistance. Once infected, plants never recovered, regardless of transgene translatability. Transgenic plants exhibiting a range of resistance levels were examined for transgene copy number, mRNA and protein levels. Although ribonuclease protection assays demonstrated that transgene mRNA levels were very low, resistant lines had consistently more steady-state transgene mRNA than susceptible lines. Furthermore, chlorotic or necrotic local lesions developed on the inoculated leaves of transgenic lines containing translatable transgenes, but not on inoculated leaves of lines containing nontranslatable transgenes. These results demonstrate that translatability of the transgene and possibly expression of the transgene protein itself facilitates replicase-mediated resistance to CMV in tobacco. PMID- 10675397 TI - Detection of beet yellows closterovirus methyltransferase-like and helicase-like proteins in vivo using monoclonal antibodies. AB - In the positive-stranded RNA genome of beet yellows closterovirus (BYV), the 5' terminal ORF 1a encodes a 295 kDa polyprotein with the domains of papain-like cysteine proteinase, methyltransferase (MT) and helicase (HEL), whereas ORF 1b encodes an RNA-dependent RNA polymerase. Eleven and five hybridoma cell lines secreting monoclonal antibodies (MAbs) were derived from mice injected with the bacterially expressed fragments of the BYV 1a product encompassing the MT and HEL domains, respectively. On immunoblots of protein from BYV-infected Tetragonia expansa plants, four MAbs against the MT recognized a approximately 63 kDa protein, and two MAbs against the HEL recognized a approximately 100 kDa protein. Both the methyltransferase-like protein and the helicase-like protein were found mainly in the fractions of large organelles (P1) and membranes (P30) of the infected plants. These data clearly indicate that (i) the BYV methyltransferase like and helicase-like proteins, like other related viral enzymes, are associated with membrane compartments in cells, and (ii) the 1a protein, apart from the cleavage by the leader papain-like proteinase that is expected to produce the 66 kDa and 229 kDa fragments, undergoes additional processing by a virus-encoded or cellular proteinase. PMID- 10675398 TI - Nucleotide sequence and organization of ten open reading frames in the genome of grapevine leafroll-associated virus 1 and identification of three subgenomic RNAs. AB - The genome of Grapevine leafroll-associated virus 1 (GLRaV-1) was cloned and the sequence of 12394 nts determined. It contains 10 major open reading frames (ORFs) and a 3'-non-coding region lacking a poly(A) tract. The first ORF (ORF 1a) encodes a putative RNA helicase at the C-terminal portion of an apparently larger protein. The downstream ORF, 1b, overlaps ORF 1a and lacks an initiation codon. This ORF encodes an RNA-dependent RNA polymerase of M(r) 59276. ORF 2 encodes a small hydrophobic protein of M(r) 6736, and ORF 3 encodes a homologue of the HSP70 family of heat shock proteins and has an M(r) of 59500. ORF 4 encodes a protein with an M(r) of 54648 that shows similarity to the corresponding proteins of other closteroviruses. ORF 5 encodes the viral coat protein (CP) with an M(r) of 35416. The identity of this ORF as the CP gene was confirmed by expression in Escherichia coli and testing with the viral antibody. ORFs 6 and 7 code for two CP-related products with M(r) of 55805 and 50164, respectively. ORFs 8 and 9 encode proteins of M(r) 21558 and 23771 with unknown functions. Using DNA probes to different regions of the GLRaV-1 sequence, three major 3'-coterminal subgenomic RNA species were identified and mapped on the GLRaV-1 genome. Phylogenetic analyses of the individual genes of GLRaV-1 demonstrated a closer relationship between GLRaV-1 and GLRaV-3 than with other closteroviruses. PMID- 10675399 TI - Tagging potato leafroll virus with the jellyfish green fluorescent protein gene. AB - A full-length cDNA corresponding to the RNA genome of Potato leafroll virus (PLRV) was modified by inserting cDNA that encoded the jellyfish green fluorescent protein (GFP) into the P5 gene near its 3' end. Nicotiana benthamiana protoplasts electroporated with plasmid DNA containing this cDNA behind the 35S RNA promoter of Cauliflower mosaic virus became infected with the recombinant virus (PLRV-GFP). Up to 5% of transfected protoplasts showed GFP-specific fluorescence. Progeny virus particles were morphologically indistinguishable from those of wild-type PLRV but, unlike PLRV particles, they bound to grids coated with antibodies to GFP. Aphids fed on extracts of these protoplasts transmitted PLRV-GFP to test plants, as shown by specific fluorescence in some vascular tissue and epidermal cells and subsequent systemic infection. In plants agroinfected with PLRV-GFP cDNA in pBIN19, some cells became fluorescent and systemic infections developed. However, after either type of inoculation, fluorescence was mostly restricted to single cells and the only PLRV genome detected in systemically infected tissues lacked some or all of the inserted GFP cDNA, apparently because of naturally occurring deletions. Thus, intact PLRV-GFP was unable to move from cell to cell. Nevertheless, PLRV-GFP has novel potential for exploring the initial stages of PLRV infection. PMID- 10675400 TI - The non-essential UL50 gene of avian infectious laryngotracheitis virus encodes a functional dUTPase which is not a virulence factor. AB - The DNA sequence of the infectious laryngotracheitis virus (ILTV) UL50, UL51 and UL52 gene homologues was determined. Although the deduced UL50 protein lacks the first of five conserved domains of the corresponding proteins of mammalian alphaherpesviruses, the ILTV gene product was also shown to possess dUTPase activity. The generation of UL50-negative ILTV mutants was facilitated by recombination plasmids encoding green fluorescent protein (GFP), and expression constructs of predicted transactivator proteins of ILTV (alphaTIF, ICP4) were successfully used to increase the infectivity of viral genomic DNA. A GFP expressing UL50-deletion mutant of ILTV showed reduced cell-to-cell spread in vitro, and was attenuated in vivo. A similar deletion mutant without the foreign gene, however, propagated like wild-type ILTV in cell culture and was pathogenic in chickens. We conclude that the viral dUTPase is not required for efficient replication of ILTV in the respiratory tract of infected animals. The replication defect of the GFP-expressing ILTV recombinant is most likely caused by toxic effects of the reporter gene product, since spontaneously occurring inactivation mutants exhibited wild-type-like growth. PMID- 10675401 TI - Resistance of herpes simplex virus type 1 against different phosphonylmethoxyalkyl derivatives of purines and pyrimidines due to specific mutations in the viral DNA polymerase gene. AB - Drug-resistant strains of herpes simplex virus type 1 (HSV-1) were selected under the pressure of (S)-3-hydroxy-2-phosphonylmethoxypropyl (HPMP) derivatives of cytosine (HPMPC, cidofovir) and adenine (HPMPA) and 2-phosphonylmethoxyethyl (PME) derivatives of adenine (PMEA, adefovir) and 2,6-diaminopurine (PMEDAP). HPMPC-resistant (HPMPC(r)) and HPMPA(r) strains were cross-resistant to one another, but they remained sensitive to foscarnet (PFA), acyclovir (ACV) and the PME derivatives, while the PMEA(r) and PMEDAP(r) strains showed cross-resistance to PFA and ACV. The PMEA(r), PMEDAP(r) and PFA(r) mutants all revealed a single nucleotide change resulting in a Ser-724 to Asn mutation within the conserved region II of the DNA polymerase. Two HPMPA(r) clones and one HPMPC(r) clone possessed single amino acid changes in the DNA polymerase (HPMPA(r) clone D1, Leu 1007 to Met; HPMPA(r) clone B5, Ile-1028 to Thr; HPMPC(r) clone C3, Val-573 to Met). The HPMPC(r) clone A4 contained two mutations, Ala-136 to Thr and Arg-700 to Met. The mutation at position 136, located outside the catalytic domain of the enzyme, was not detected in other HPMPC(r) clones, suggesting that this mutation may not be responsible for the resistant phenotype. Residue 573 is located within the 3'-->5' exonuclease editing domain close to the catalytically important residues Tyr-577 and Asp-581. Similarly, residue 700 is located in the palm subdomain of the catalytic domain, adjacent to the Asp residues 717, 886 and 888 that are vital for polymerase activity. The HPMPA(r) mutations at residues 1007 and 1028, beyond the last conserved region, still fall within the thumb subdomain of the catalytic domain. The different drug-resistant mutants varied in neurovirulent behaviour, the HPMPC(r) strains showing reduced neurovirulence compared with the wild-type. PMID- 10675402 TI - Expression from the herpes simplex virus type 1 latency-associated promoter in the murine central nervous system. AB - Herpes simplex virus type 1 (HSV-1) establishes latency in neurones of both the central (CNS) and peripheral nervous system and can be used to drive long-term expression of the lacZ reporter gene by insertion of an encephalomyocarditis virus IRES-linked gene 1.5 kb downstream of the latency-associated transcript (LAT) start site. However, the kinetics of LAT promoter (LAP) activity, and its ability to function in all neuronal types within the CNS has not been studied in detail. In order to address these issues, mice were infected via the ear pinna with 2x10(6) p.f.u. of either SC16-LbetaA, which contains an IRES-linked lacZ under the control of LAP, or SC16-C3b, which expresses LacZ under the control of the human cytomegalovirus immediate early (HCMV-IE) promoter. Three to five animals from each group were sampled over a time-course from 5 days to 1 year post-infection (p.i.), and brainstem and spinal cord sections were examined histochemically for LacZ expression. We found that HCMV-IE promoter activity could be detected within distinct CNS regions from 5 to 15 days p.i. In contrast, LAP-driven LacZ expression was first detected at 7 days p.i. and persisted for at least 1 year. At times up to 34 days p.i., LAP activity was seen in similar regions of the CNS as those which were positive for HCMV-IE promoter activity during the acute stage of infection. After 34 days, however, the numbers of cells in which the LAP was active decreased and labelled motorneurones were predominantly detected in the facial and hypoglossal nuclei and occasionally also in the ventral spinal cord. These results suggest that following the establishment of latency in the CNS, the efficiency of long-term LAP-mediated gene expression may be influenced by the neuronal cell type in which latency is established. PMID- 10675403 TI - Suppressive effects of human herpesvirus-6 on thrombopoietin-inducible megakaryocytic colony formation in vitro. AB - Two clinical observations, the association of human herpesvirus-6 (HHV-6) with delayed engraftment after stem cell transplantation and thrombocytopenia concomitant with exanthema subitum, prompted us to evaluate the suppressive effects of HHV-6 on thrombopoiesis in vitro. Different culture conditions for thrombopoietin (TPO)-inducible colonies in semi-solid matrices were examined. Using cord blood mononuclear cells as the source of haematopoietic progenitors, two types of colonies, megakaryocyte colony-forming units (CFU-Meg) and non-CFU Meg colonies, were established. The former colonies were identified by the presence of cells with translucent cytoplasm and highly refractile cell membrane, most of which were positive for the CD41 antigen. Although the plating efficiency of both types was much higher under serum-containing conditions than under serum free conditions, the proportion of CFU-Meg to non-CFU-Meg colonies was consistently higher under serum-free conditions. The plating efficiency of CFU Meg colonies was doubled by adding stem cell factor to the serum-free matrix. The effects of two variants of HHV-6 (HHV-6A and 6B) and human herpesvirus-7 (HHV-7) on TPO-inducible colonies were then compared. HHV-6B inhibited both CFU-Meg and non-CFU-Meg colony formation under serum-free and serum-containing conditions. HHV-6A had similar inhibitory effects. In contrast, HHV-7 had no effect on TPO inducible colony formation. Heat-inactivation and ultra-filtration of the virus sample completely abolished the suppressive effect. After infection of CD34(+) cells with HHV-6, the viral genome was consistently detected by in situ hybridization. These data suggest that the direct effect of HHV-6 on haematopoietic progenitors is one of the major causes of the suppression of thrombopoiesis. PMID- 10675404 TI - Morphogenesis and release of fowlpox virus. AB - Release of fowlpox virus (FWPV) as extracellular enveloped virus (EEV) appears to proceed both by the budding of intracellular mature virus (IMV) through the plasma membrane and by the fusion of intracellular enveloped virus (IEV) with the plasma membrane. Based on the frequency of budding events compared to wrapping events observed by electron microscopy, FWPV FP9 strain seems to exit chick embryo fibroblast cells predominantly by budding. In contrast to vaccinia virus (VV), the production of FWPV extracellular virus particles is not affected by N(1)-isonicotinoyl-N(2)-3-methyl-4-chlorobenzoylhydrazine (IMCBH). Comparison of the sequence of the VV F13L gene product with its FWPV orthologue showed a mutation, in the fowlpox protein, at the residue involved in IMCBH resistance in a mutant VV. Glucosamine, monensin or brefeldin A did not have any specific effect on FWPV extracellular virus production. Cytochalasin D, which inhibits the formation of actin filaments, reduces the production of extracellular virus particles by inhibiting the release of cell-associated enveloped virus (CEV) particles from the plasma membrane. Involvement of actin filaments in this mechanism is further supported by the co-localization of actin with viral particles close to the plasma membrane in the absence of cytochalasin D. Actin is also co-localized with virus factories. PMID- 10675405 TI - Binding of bovine papillomavirus type 4 E8 to ductin (16K proteolipid), down regulation of gap junction intercellular communication and full cell transformation are independent events. AB - The E8 open reading frame of bovine papillomavirus type 4 encodes a small hydrophobic polypeptide that contributes to primary cell transformation by conferring to cells the ability to form foci and to grow in low serum and in suspension. Wild-type E8 binds in vitro to ductin, a component of gap junctions, and this binding is accompanied by a loss of gap junction intercellular communication in transformed bovine fibroblasts. However, through the analysis of a panel of E8 mutants, we show here that binding of E8 to ductin is not sufficient for down-regulation of gap junction communication and that there is no absolute correlation between down-regulation of gap junction communication and the transformed phenotype. PMID- 10675406 TI - The effects of interferon on the expression of human papillomavirus oncogenes. AB - The effects of interferon (IFN)-alpha, IFN-beta and IFN-gamma on human papillomavirus (HPV) oncogene expression were studied in various cervical carcinoma cell lines containing integrated copies of either HPV type 16 or HPV type 18. The levels of E6 and E7 transcripts were examined 6 h and 30 h after treatment with IFN. In HeLa cells, all three classes of IFNs effected a decrease in the level of HPV-18 E6 and E7 transcripts. On the other hand, none of the IFNs altered the level of these transcripts in C-4II cells. Only IFN-gamma decreased the level of HPV-16 E6 and E7 transcripts in CaSki and HPK1A cells, while IFN gamma actually increased the level of these transcripts in SiHa cells. This differential IFN regulation of HPV expression in various cervical cancer cell lines may account for the contradictory clinical results observed after treatment of cervical cancer with IFN. PMID- 10675407 TI - Specific serum IgG, IgM and IgA antibodies to human papillomavirus types 6, 11, 16, 18 and 31 virus-like particles in human immunodeficiency virus-seropositive women. AB - To evaluate the humoral immune response to human papillomavirus (HPV) in women infected with human immunodeficiency virus (HIV), serum samples of 83 HIV positive individuals were analysed by ELISA for specific antibodies of the isotypes IgG, IgA and IgM recognizing HPV-6, -11, -16, -18 and -31 L1 virus-like particles (VLPs). Papillomavirus-related lesions were present in 30 of 83 HIV positive women. Twenty-one women (25%) presented with high-/intermediate-grade anogenital squamous intraepithelial lesions. PCR analysis and sequencing for HPV typing was done from biopsy specimens of 18 women; PCR-positive results were obtained in 90% of cases. In addition, HPV DNA hybrid capture assays were performed from cervical swabs of 58 HIV-positive women, 53% of whom had a positive result for high-risk HPV. Overall, positive IgG reactivity to HPV-6/-11 and HPV-16/-18/-31 was seen in 19%/31% and 49%/30%/24% of HIV-positive women, respectively. HPV-seropositivity was even higher than in 48 HIV-negative cervical intraepithelial neoplasia/cancer patients with percentages as follows: 8%/2% and 31%/15%/15%. This difference was significant for HPV-16 (P=0.046). IgA responses were comparable to IgG. IgM responses were low. The extraordinarily high rate of antibodies to the capsid protein L1 of high-risk HPVs (HPV-16, -18 and/or -31) in 58% of HIV-positive women compared to 19% (P=0.00001) of 102 healthy HIV-negative control women suggests a high lifetime cumulative exposure to HPV and increased expression of capsid proteins due to cellular immunodeficiency in HIV-infected women. PMID- 10675408 TI - Development of a genetically marked recombinant rinderpest vaccine expressing green fluorescent protein. AB - In order to effectively control and eliminate rinderpest, a method is required to allow serological differentiation between animals that have been vaccinated and those which have recovered from natural infection. One way of doing this would be to engineer the normal vaccine to produce a genetically marked rinderpest virus (RPV) vaccine. We constructed two modified cDNA clones of the RPV RBOK vaccine strain with the coding sequence of the green fluorescent protein (GFP) gene inserted as a potential genetic marker. RPVINS-GFP virus was designed to produce independent and high level expression of GFP inside infected cells, whilst the GFP expressed by RPVSIG-GFP virus was designed to be efficiently secreted. Infectious recombinant virus was rescued in cell culture from both constructs. The effectiveness of these viruses in stimulating protective immunity and antibody responses to the marker protein was tested by vaccination of cattle and goats. All of the vaccinated animals were completely protected when challenged with virulent virus: RPV in cattle or peste-des-petits ruminants virus in the goats. ELISA showed that all of the animals produced good levels of anti-RPV antibodies. Three of the four cattle and the two goats vaccinated with RPVSIG-GFP produced detectable levels of anti-GFP antibodies. In contrast, no anti-GFP antibodies were produced in the four cattle and two goats vaccinated with RPVINS GFP. Therefore, secretion of the GFP marker protein was absolutely required to elicit an effective humoral antibody response to the marker protein. PMID- 10675409 TI - An amino acid in the heptad repeat 1 domain is important for the haemagglutinin neuraminidase-independent fusing activity of simian virus 5 fusion protein. AB - A canine isolate (strain T1) of simian virus 5 (SV-5) performed multiple replication in BHK cells but did not induce cell fusion for up to 3 days. In contrast, a prototype strain (WR) provoked extensive cell fusion within 2 days during the course of its replication. Accordingly, the fusion (F) protein of the T1 strain did not cause cell fusion even when co-expressed with the SV-5 haemagglutinin-neuraminidase (HN) protein, whereas the WR F protein induced cell fusion in the presence of the HN protein. Differences in the predicted amino acid sequences of the T1 and WR F proteins were found at 12 positions and it was proved that the T1 F protein had a longer cytoplasmic tail than the WR F protein. By reducing the length of the cytoplasmic tail or by replacing the tail with the WR F counterpart, the T1 F protein partly restored its HN-dependent fusing activity. Chimeric and mutational analyses between the T1 F protein and the mutant F protein (L22P) suggested that Glu-132 in the heptad repeat 1 domain was involved in the HN-independent fusing activity in addition to the previously identified Pro-22 at the F(2) N terminus. It was also shown that Ala-290 in the heptad repeat 3 domain contributed to the HN-independent fusing activity to some extent. PMID- 10675410 TI - Neutralization of measles virus wild-type isolates after immunization with a synthetic peptide vaccine which is not recognized by neutralizing passive antibodies. AB - The sequence H379-410 of the measles virus haemagglutinin (MV-H) protein forms a surface-exposed loop and contains three cysteine residues (Cys-381, Cys-386 and Cys-394) which are conserved among all measles isolates. It comprises the minimal sequential B cell epitope (BCE) (H386-400) of the neutralizing and protective MAb BH6 that neutralizes all wild-type viruses tested. The aim of this study was to design synthetic peptides which induce neutralizing antibodies against MV wild type isolates. Peptides containing one or two copies of T cell epitopes (TCE) and BCEs of different lengths (H386-400, B(CC); H379-400, B(CCC)), in different combinations and orientations were produced and iteratively optimized for inducing neutralizing antibodies. Peptides with the shorter BCE induced sera that cross-reacted with MV but did not neutralize. The longer BCE containing the three cysteines (B(CCC)) and two homologous TCE were required for neutralization activity. These sera neutralized wild-type strains of different clades and geographic origins. Neutralizing serum was also obtained after immunization with human promiscuous TCEs. Furthermore B(CCC)-based peptides were fully immunogenic even in the presence of pre-existing MV-specific antibodies. The results suggest that subunit vaccines based on such peptides could potentially be used to actively protect infants against wild-type viruses irrespective of persisting maternal antibodies. PMID- 10675411 TI - Reduced levels of neuraminidase of influenza A viruses correlate with attenuated phenotypes in mice. AB - We have previously obtained four transfectant influenza A viruses containing neuraminidase (NA) genes with mutated base pairs in the conserved double-stranded RNA region of the viral promoter by using a ribonucleoprotein transfection system. Two mutant viruses (D2 and D1/2) which share a C-G-->A-U mutation at positions 11 and 12 of the 3' and 5' ends, respectively, of the NA gene, showed an approximate 10-fold reduction of NA-specific mRNA and protein levels (Fodor et al., Journal of Virology 72, 6283-6290, 1998). These viruses have now allowed us to determine the effects of decreased NA levels on virus pathogenicity. Both D2 and D1/2 viruses were highly attenuated in mice, and their replication in mouse lungs was highly compromised as compared with wild-type influenza A/WSN/33 virus. The results highlight the importance of the level of NA activity in the biological cycle and virulence of influenza viruses. Importantly, mice immunized by a single intranasal administration of 10(3) infectious units of D2 or D1/2 viruses were protected against challenge with a lethal dose of wild-type influenza virus. Attenuation of influenza viruses by mutations resulting in the decreased expression of a viral protein represents a novel strategy which could be considered for the generation of live attenuated influenza virus vaccines. PMID- 10675412 TI - Nucleotide sequences and phylogeny of the nucleocapsid gene of Oropouche virus. AB - The nucleotide sequence of the S RNA segment of the Oropouche (ORO) virus prototype strain TRVL 9760 was determined and found to be 754 nucleotides in length. In the virion-complementary orientation, the RNA contained two overlapping open reading frames of 693 and 273 nucleotides that were predicted to encode proteins of 231 and 91 amino acids, respectively. Subsequently, the nucleotide sequences of the nucleocapsid genes of 27 additional ORO virus strains, representing a 42 year interval and a wide geographical range in South America, were determined. Phylogenetic analyses revealed that all the ORO virus strains formed a monophyletic group that comprised three distinct lineages. Lineage I contained the prototype strain from Trinidad and most of the Brazilian strains, lineage II contained six Peruvian strains isolated between 1992 and 1998, and two strains from western Brazil isolated in 1991, while lineage III comprised four strains isolated in Panama during 1989. PMID- 10675413 TI - Recombinant Semliki Forest virus particles expressing louping ill virus antigens induce a better protective response than plasmid-based DNA vaccines or an inactivated whole particle vaccine. AB - Louping ill virus (LIV) infection of mice was used as a model to evaluate the protective efficacy of Semliki Forest virus (SFV)-based vaccines in comparison to a standard DNA vaccine and a commercial chemically inactivated vaccine. The recombinant SFV-based vaccines consisted of suicidal particles and a naked layered DNA/RNA construct. The nucleic acid vaccines expressed the spike precursor prME and the nonstructural protein 1 (NS1) antigens of LIV. Three LIV strains of graded virulence for mice were used for challenge. One of these was a naturally occurring antibody escape variant. All vaccines tested induced humoral immunity but gave varying levels of protection against lethal challenge. Only recombinant SFV particles administered twice gave full protection against neuronal degeneration and encephalitis induced by two of the three challenge strains, and partial protection against the highly virulent strain, whereas the other vaccines tested gave lower levels of partial protection. PMID- 10675414 TI - Biochemical characterization of a hepatitis C virus RNA-dependent RNA polymerase mutant lacking the C-terminal hydrophobic sequence. AB - The RNA-dependent RNA polymerase activity of hepatitis C virus is carried out by the NS5B protein. The full-length protein was previously purified as a non-fusion protein from insect cells infected with a recombinant baculovirus. The characterization is now described of a C-terminal hydrophobic domain deletion mutant of NS5B purified from E. coli. In addition to increased solubility, deletion of this sequence also positively affected the polymerase enzymatic activity. The efficiency of nucleotide polymerization of both the full-length and the C-terminal truncated enzymes were compared on homopolymeric template-primer couples as well as on RNA templates with heteropolymeric sequences. The largest difference in the polymerase activity was observed on the latter. On all the templates, the increased activity could be ascribed, at least in part, to enhanced template turnover of the deletion mutant with respect to the full-length enzyme. The elongation rates of the two enzyme forms were compared under single processive cycle conditions. Under these conditions, both the full-length and the deletion mutant were able to incorporate about 700 nt/min. PMID- 10675415 TI - Phylogenetic analysis of GBV-C/hepatitis G virus. AB - Comparison of 33 epidemiologically distinct GBV-C/hepatitis G virus complete genome sequences suggests the existence of four major phylogenetic groupings that are equally divergent from the chimpanzee isolate GBV-C(tro) and have distinct geographical distributions. These four groupings are not consistently reproduced by analysis of the virus 5'-noncoding region (5'-NCR), or of individual genes or subgenomic fragments with the exception of the E2 gene as a whole or of 200-600 nucleotide fragments from its 3' half. This region is upstream of a proposed anti sense reading frame and contains conserved potential RNA secondary structures that may be capable of directing the internal initiation of translation. Phylogenetic analysis of this region from certain South African isolates is consistent with previous analysis of the 5'-NCR suggesting that these belong to a fifth group. The geographical distribution of virus variants is consistent with a long evolutionary history that may parallel that of pre-historic human migrations, implying that the long-term evolution of this RNA virus is extremely slow. PMID- 10675416 TI - Phylogeny of the genus flavivirus using complete coding sequences of arthropod borne viruses and viruses with no known vector. AB - Attempts to define the evolutionary relationships and origins of viruses in the genus Flavivirus are hampered by the lack of genetic information particularly amongst the non-vectored flaviviruses. Using a novel protocol for sequence determination, the first complete coding sequence of St Louis encephalitis virus and those of two representative non-vectored flaviviruses, Rio Bravo (isolated from bat) and Apoi (isolated from rodent), are reported. The encoded polyproteins of Rio Bravo and Apoi virus are the smallest described to date within the genus FLAVIVIRUS: The highest similarities with other flaviviruses were found in the NS3 and NS5 genes. The proteolytic cleavage sites for the viral serine protease were highly conserved among the flaviviruses completely sequenced to date. Comparative genetic amino acid alignments revealed that p-distance cut-off values of 0.330-0.470 distinguished the arthropod-borne viruses according to their recognized serogroups and Rio Bravo and Apoi virus were assigned to two distinct non-vectored virus groups. Within these serogroups, cladogenesis based on the complete ORF sequence was similar to trees based on envelope and NS5 sequences. In contrast, branching patterns at the deeper nodes of the tree were different from those reported in the previous study of NS5 sequences. The significance of these observations is discussed. PMID- 10675417 TI - Leader switching occurs during the rescue of defective RNAs by heterologous strains of the coronavirus infectious bronchitis virus. AB - A defective RNA (D-RNA), CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was rescued (replicated and packaged) using four heterologous strains of IBV as helper virus. Sequence analysis of the genomic RNA from the four heterologous IBV strains (M41, H120, HV10 and D207) identified nucleotide differences of up to 17% within the leader sequence and up to 4.3% within the whole of the adjacent 5' untranslated region (UTR). Analysis of the 5' ends of the rescued D-RNAs showed that the Beaudette leader sequence, present on the initial CD-61, had been replaced with the corresponding leader sequence from the helper IBV strain but the adjacent 5' UTR sequence of the rescued D-RNAs corresponded to the original CD-61 Beaudette sequence. These results demonstrated that the phenomenon of leader switching previously identified for the coronaviruses murine hepatitis virus and bovine coronavirus (BCoV) also occurred during the replication of IBV D-RNAs. Three predicted stem-loop structures were identified within the 5' UTR of IBV. Stem loop I showed a high degree of covariance amongst the IBV strains providing phylogenetic evidence that this structure exists and is potentially involved in replication, supporting previous observations that a BCoV stem-loop homologue was essential for replication of BCoV defective interfering RNAs. PMID- 10675418 TI - Molecular epidemiology of coxsackievirus B4 and disclosure of the correct VP1/2A(pro) cleavage site: evidence for high genomic diversity and long-term endemicity of distinct genotypes. AB - Genetic diversity among 107 coxsackievirus B4 field isolates has been studied. These isolates included clinical and environmental isolates originating from Finland, the Netherlands and France, and also from several other countries, including the USA. Three genomic regions were used for phylogenetic analyses: the VP1/2A junction, the entire VP1 and the VP4/VP2 region. Alignment of the deduced amino acid sequence in the VP1/2A junction revealed extensive sequence variation at the previously proposed cleavage site. MS analysis of proteolytic fragments from VP1 revealed that the exact cleavage site is situated between amino acid residues Thr-849 and Gly-850. At least seven distinct genetic lineages, or genotypes, had been circulating in Europe during the period 1959-1998. Two genotypes were endemic in the Netherlands during most of the investigated period. Genetically closely related strains could be found in different countries, and different genotypes co-circulated at the same time in a given country. Clustering patterns were identical in the three genomic intervals. In the VP4/VP2 region, the intraserotypic variation approached interserotype variation. Sequence comparisons of the entire VP1 gene gave a reliable genetic identification of enterovirus serotype. It is suggested that, for genotype classification of previously serotyped coxsackievirus B4 isolates, comparison of VP1/2A sequences is sufficient, but for more detailed investigation of genetic relationships, and for 'genetic serotyping', the entire VP1 gene should be used. The VP4/VP2 region is less reliable for genetic serotyping and genotyping, although the primers are able to amplify many different serotypes. PMID- 10675419 TI - Biochemical characterization of salmon pancreas disease virus. AB - Salmon pancreas disease virus (SPDV) has been shown to cause severe economic losses in farmed Atlantic salmon (Salmo salar) and has been reported to occur in Europe, Scandinavia and the United States. This paper describes the biochemical characterization of SPDV in terms of its RNA and protein composition. SPDV was purified by precipitation from infected Chinook salmon embryo (CHSE-214) cell culture supernatant and sucrose density-gradient centrifugation. Fractions containing virus were identified by an immunodot blot assay using an SPDV specific MAb. Two major proteins with molecular masses of approximately 55 and 50 kDa, putatively identified as the E1 and E2 alphavirus glycoproteins respectively, were detected when purified virus preparations were analysed by PAGE. Radiolabelling experiments indicated that SPDV infection of CHSE-214 cells did not shut-off host-cell protein synthesis, making attempts to identify virus specific proteins unsuccessful. However, radioimmunoprecipitation assay (RIPA) experiments showed that two SPDV-specific MAbs reacted with a protein in the 50 55 kDa range. Northern blot hybridization with cloned cDNA probes indicated that infected cells contained RNA species of approximately 11.4 and 4 kb, which correspond to the genomic and subgenomic RNAs specified by SPDV. The results described are consistent with SPDV being characterized as an alphavirus. PMID- 10675420 TI - The C-terminal domain of rotavirus NSP5 is essential for its multimerization, hyperphosphorylation and interaction with NSP6. AB - Rotavirus NSP5 is a non-structural phosphoprotein with putative autocatalytic kinase activity, and is present in infected cells as various isoforms having molecular masses of 26, 28 and 30-34 kDa. We have previously shown that NSP5 forms oligomers and interacts with NSP6 in yeast cells. Here we have mapped the domains of NSP5 responsible for these associations. Deletion mutants of the rotavirus YM NSP5 were constructed and assayed for their ability to interact with full-length NSP5 and NSP6 using the yeast two-hybrid assay. The homomultimerization domain was mapped to the 20 C-terminal aa of the protein, which have a predicted alpha-helical structure. A deletion mutant lacking the 10 C-terminal aa (DeltaC10) failed to multimerize both in yeast cells and in an in vitro affinity assay. When transiently expressed in MA104 cells, NSP5 became hyperphosphorylated (30-34 kDa isoforms). In contrast, the DeltaC10 mutant produced forms equivalent to the 26 and 28 kDa species, but was poorly hyperphosphorylated, suggesting that multimerization is important for this proposed activity of the protein. The interaction domain with NSP6 was found to be present in the 35 C-terminal aa of NSP5, overlapping the multimerization domain of the protein, and suggesting that NSP6 might have a regulatory role in the self-association of NSP5. NSP6 was also found to interact with wild-type NSP5, but not with its mutant DeltaC10, in cells transiently transfected with plasmids encoding these proteins, confirming the relevance of the 10 C-terminal aa for the formation of the heterocomplex. PMID- 10675421 TI - Identification and differentiation of the nine African horse sickness virus serotypes by RT-PCR amplification of the serotype-specific genome segment 2. AB - This paper describes the first RT-PCR for discrimination of the nine African horse sickness virus (AHSV) serotypes. Nine pairs of primers were designed, each being specific for one AHSV serotype. The RT-PCR was sensitive and specific, providing serotyping within 24 h. Perfect agreement was recorded between the RT PCR and virus neutralization for a coded panel of 56 AHSV reference strains and field isolates. Serotyping was achieved successfully with live and formalin inactivated AHSVs, with isolates of virus after low and high passage through either tissue culture or suckling mouse brain, with viruses isolated from widely separated geographical areas and with viruses isolated up to 37 years apart. Overall, this RT-PCR provides a rapid and reliable method for the identification and differentiation of the nine AHSV serotypes, which is vital at the start of an outbreak to enable the early selection of a vaccine to control the spread of disease. PMID- 10675422 TI - Subterminal viral DNA nucleotides as specific recognition signals for human immunodeficiency virus type 1 and visna virus integrases under magnesium dependent conditions. AB - Many reports describe the characteristics of susceptible viral DNA substrates to various retroviral integrases during in vitro reactions in which manganese serves as the divalent cation cofactor for site-specific nicking. However, manganese is known to alter the specificity of some endonucleases and magnesium may be the divalent cation used during retroviral integration in vivo. To address these concerns, we identified conditions under which the integrases of human immunodeficiency virus type 1 and visna virus were optimally active with magnesium (the first time such activity was shown for visna virus integrase) and used these conditions to test the susceptibility of a series of oligodeoxynucleotide substrates. The data show that two base pairs immediately internal to the conserved CA dinucleotide near the termini of retroviral DNA are selectively recognized by the two integrases and that the final six base pairs of viral DNA contain sufficient sequence information for specific recognition and cleavage by each enzyme. The results validate the importance of the subterminal viral DNA positions even in the presence of magnesium and identify viral DNA positions that functionally interact with integrase. The data obtained under magnesium-dependent conditions, which were obtained with substrates containing single and multiple base-pair substitutions and two different retroviral integrases, are consistent with those previously obtained with manganese. Thus, the large body of manganese-dependent data identifying terminal viral DNA positions that are important in substrate recognition by various integrases likely reflects interactions that are biologically relevant. PMID- 10675423 TI - Molecular analysis of ovine prion protein identifies similarities between BSE and an experimental isolate of natural scrapie, CH1641 PMID- 10675424 TI - The effect of weight loss in overweight, lactating women on the growth of their infants. AB - BACKGROUND: The retention of weight gained during pregnancy may contribute to obesity. Lactation should promote weight loss, but weight loss is highly variable among lactating women. The risks associated with the restriction of energy intake during lactation have not been adequately evaluated. The purpose of this study was to determine whether weight loss by women during lactation affects the growth of their infants. METHODS: We randomly assigned 40 breast-feeding women who were overweight (defined as a body-mass index [the weight in kilograms divided by the square of the height in meters] of 25 to 30) at 4 weeks post partum either to restrict their energy intake by 500 kcal per day and to exercise for 45 minutes per day for 4 days per week (the diet-and-exercise group) or to maintain their usual dietary intake and not exercise more than once per week for 10 weeks (the control group). We measured the weight and fat mass of the women and the weight and length of the infants before, during, and at the end of the study period. RESULTS: The mean (+/-SD) energy intake decreased by 544+/-471 kcal per day in the diet-and-exercise group. As compared with the control group, the women in the diet-and-exercise group lost more weight (4.8+/-1.7 kg vs. 0.8+/-2.3 kg, P<0.001) and fat mass (4.0+/-2.0 kg vs. 0.3+/-1.8 kg, P<0.001). The gains in weight and length of the infants whose mothers were in the diet-and-exercise group (1925+/ 500 g and 7.8+/-2.0 cm, respectively) were not significantly different from those of the infants whose mothers were in the control group (1861+/-576 g and 7.3+/ 1.7 cm). CONCLUSIONS: Weight loss of approximately 0.5 kg per week between 4 and 14 weeks post partum in overweight women who are exclusively breast-feeding does not affect the growth of their infants. PMID- 10675425 TI - Effect of exercise on coronary endothelial function in patients with coronary artery disease. AB - BACKGROUND: Studies of the cardioprotective effects of exercise training in patients with coronary artery disease have yielded contradictory results. Exercise training has been associated with improvement in myocardial perfusion even in patients who have progression of coronary atherosclerosis. We therefore conducted a prospective study of the effect of exercise training on endothelial function in patients with coronary artery disease. METHODS: We randomly assigned 19 patients with coronary endothelial dysfunction, indicated by abnormal acetylcholine-induced vasoconstriction, to an exercise-training group (10 patients) or a control group (9 patients). To reduce confounding, patients with coronary risk factors that could be influenced by exercise training (such as diabetes, hypertension, hypercholesterolemia, and smoking) were excluded. In an initial study and after four weeks, the changes in vascular diameter in response to the intracoronary infusion of increasing doses of acetylcholine (0.072, 0.72, and 7.2 microg per minute) were assessed. The mean peak flow velocity was measured by Doppler velocimetry, and the diameter of epicardial coronary vessels was measured by quantitative coronary angiography. RESULTS: In the initial study, the two groups had similar vasoconstrictive responses to acetylcholine. After four weeks of exercise training, coronary-artery constriction in response to acetylcholine at a dose of 7.2 microg per minute was reduced by 54 percent (from a mean [+/-SE] decrease in the luminal diameter of 0.41+/-0.05 mm in the initial study to a decrease of 0.19+/-0.07 mm at four weeks; P<0.05 for the comparison with the change in the control group). In the exercise-training group, the increases in mean peak flow velocity in response to 0.072, 0.72, and 7.2 microg of acetylcholine per minute were 12+/-7, 36+/-11, and 78+/-16 percent, respectively, in the initial study. After four weeks of exercise, the increases in response to acetylcholine were 27+/-7, 73+/-19, and 142+/-28 percent (P<0.01 for the comparison with the control group). Coronary blood-flow reserve (the ratio of the mean peak flow velocity after adenosine infusion to the resting velocity) increased by 29 percent after four weeks of exercise (from 2.8+/-0.2 in the initial study to 3.6+/-0.2 after four weeks; P<0.01 for the comparison with the control group). CONCLUSIONS: Exercise training improves endothelium-dependent vasodilatation both in epicardial coronary vessels and in resistance vessels in patients with coronary artery disease. PMID- 10675426 TI - A comparison of continuous thalamic stimulation and thalamotomy for suppression of severe tremor. AB - BACKGROUND: Deep-brain stimulation through an electrode implanted in the thalamus was developed as an alternative to thalamotomy for the treatment of drug resistant tremor. Stimulation is thought to be as effective as thalamotomy but to have fewer complications. We examined the effects of these two procedures on the functional abilities of patients with drug-resistant tremor due to Parkinson's disease, essential tremor, or multiple sclerosis. METHODS: Sixty-eight patients (45 with Parkinson's disease, 13 with essential tremor, and 10 with multiple sclerosis) were randomly assigned to undergo thalamotomy or thalamic stimulation. The primary outcome measure was the change in functional abilities six months after surgery, as measured by the Frenchay Activities Index. Scores for this index can range from 0 to 60, with higher scores indicating better function. Secondary outcome measures were the severity of tremor, the number of adverse effects, and patients' assessment of the outcome. RESULTS: Functional status improved more in the thalamic-stimulation group than in the thalamotomy group, as indicated by increases in the score for the Frenchay Activities Index (from 31.4 to 36.3 and from 32.0 to 32.5, respectively; difference between groups, 4.4 points; 95 percent confidence interval, 2.0 to 6.9). After adjustment for base line characteristics, multivariate analysis also showed that the thalamic stimulation group had greater improvement (difference between groups, 5.1 points; 95 percent confidence interval, 2.3 to 7.9). Tremor was suppressed completely or almost completely in 27 of 34 patients in the thalamotomy group and in 30 of 33 patients in the thalamic-stimulation group. One patient in the thalamic stimulation group died perioperatively after an intracerebral hemorrhage. With the exception of this incident, thalamic stimulation was associated with significantly fewer adverse effects than thalamotomy. Functional status was reported as improved by 8 patients in the thalamotomy group, as compared with 18 patients in the thalamic-stimulation group (P=0.01). CONCLUSIONS: Thalamic stimulation and thalamotomy are equally effective for the suppression of drug resistant tremor, but thalamic stimulation has fewer adverse effects and results in a greater improvement in function. PMID- 10675427 TI - Low-dose nitric oxide therapy for persistent pulmonary hypertension of the newborn. Clinical Inhaled Nitric Oxide Research Group. AB - BACKGROUND: Inhaled nitric oxide improves gas exchange in neonates, but the efficacy of low-dose inhaled nitric oxide in reducing the need for extracorporeal membrane oxygenation has not been established. METHODS: We conducted a clinical trial to determine whether low-dose inhaled nitric oxide would reduce the use of extracorporeal membrane oxygenation in neonates with pulmonary hypertension who were born after 34 weeks' gestation, were 4 days old or younger, required assisted ventilation, and had hypoxemic respiratory failure as defined by an oxygenation index of 25 or higher. The neonates who received nitric oxide were treated with 20 ppm for a maximum of 24 hours, followed by 5 ppm for no more than 96 hours. The primary end point of the study was the use of extracorporeal membrane oxygenation. RESULTS: Of 248 neonates enrolled, 126 were randomly assigned to the nitric oxide group and 122 to the control group. Extracorporeal membrane oxygenation was used in 78 neonates in the control group (64 percent) and in 48 neonates in the nitric oxide group (38 percent) (P=0.001). The 30-day mortality rate in the two groups was similar (8 percent in the control group and 7 percent in the nitric oxide group). Chronic lung disease developed less often in neonates treated with nitric oxide than in those in the control group (7 percent vs. 20 percent, P=0.02). The efficacy of nitric oxide was independent of the base-line oxygenation index and the primary pulmonary diagnosis. CONCLUSIONS: Inhaled nitric oxide reduces the extent to which extracorporeal membrane oxygenation is needed in neonates with hypoxemic respiratory failure and pulmonary hypertension. PMID- 10675428 TI - Images in clinical medicine. Cytomegalovirus esophageal ulcers. PMID- 10675429 TI - Gas embolism. PMID- 10675430 TI - Age-related macular degeneration. PMID- 10675431 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 5-2000. A 35-year-old man with a painful abdominal mass and fever. PMID- 10675432 TI - Dieting and exercise in overweight, lactating women. PMID- 10675433 TI - Exercise--toning up the endothelium? PMID- 10675434 TI - New treatment options for tremors. PMID- 10675436 TI - Correction: A Preliminary Study of Long-Term Treatment with Interferon Gamma-1b and Low-Dose Prednisolone in Patients with Idiopathic Pulmonary Fibrosis. PMID- 10675435 TI - Pharmacologic paralysis and withdrawal of mechanical ventilation at the end of life. PMID- 10675437 TI - Correction: A Comparison of Virtual and Conventional Colonoscopy for the Detection of Colorectal Polyps. PMID- 10675438 TI - Meeting of the Australian Microbeam Analysis Society, University of Sydney, February 16-19, 1999: Introduction. PMID- 10675439 TI - Radiofrequency Gaseous Detection Device. AB - A radiofrequency gaseous detection device is proposed for use with instruments employing charged particle beams, such as electron microscopes and ion beam technologies, as well as for detection of ionizing radiations as in proportional counters. An alternating (oscillating) electromagnetic field in the radiofrequency range is applied in a gaseous environment of the instrument. Both the frequency and amplitude of oscillation are adjustable. The electron or ion beam interacts with a specimen and releases free electrons in the gas. Similarly, an ionizing radiation source releases free electrons in the gas. The free electrons are acted upon by the alternating electromagnetic field and undergo an oscillatory motion resulting in multiple collisions with the gas molecules, or atoms. At sufficiently low pressures, the oscillating electrons also collide with surrounding walls. These processes result in an amplified electron signal and an amplified photon signal in a controlled discharge. The amplified signals, which are proportional to the initial number of free electrons, are collected by suitable means for further processing and analysis. PMID- 10675440 TI - Reverse Flow Pressure Limiting Aperture. AB - The reverse flow pressure limiting aperture is a device that creates and sustains a substantial gas pressure difference between two chambers connected via an aperture. The aperture is surrounded by an annular orifice leading to a third chamber. The third chamber is maintained at a relatively high pressure that forces gas to flow through the annular aperture into the first of said two chambers. The ensuing gas flow develops into a supersonic annular gas jet, the core of which is coaxial with the central aperture. A pumping action is created at the core of the jet and any gas molecules leaking through the aperture from the second chamber are entrained and forced into the first chamber, thus creating a substantial pressure difference between the first and second chamber. PMID- 10675441 TI - Development of a System to Provide Full, Real-time Remote Control of a Scanning Electron Microscope across the Second Generation Internet: The Teaching SEM. AB - The development and makeup of a real-time full remote control system for the University of Michigan, Department of Materials Science and Engineering Teaching SEM is described. The instrument was initially controlled via the campus local area Ethernet network and cable TV network. The latest implementation employs Fast Ethernet, Asynchronous Transfer Mode (ATM) networks, and moving picture experts group (MPEG) video encoding to effect the remote control via the computer network alone. Remote control demonstrations from Washington, DC, Dearborn, MI, and Emerson School, Ann Arbor, MI are described. PMID- 10675442 TI - Fiber-optic Based Spectral Cathodoluminescence: Simple and Economic Option for Use in Conventional and Environmental Scanning Electron Microscopy. AB - A fiber-optic based spectral cathodoluminescence (CL) system has been successfully installed on a variable pressure "environmental" scanning electron microscope (ESEM). The fine size of the fiber (50 um) has been found to require careful alignment. To provide this alignment, a simple X-Y translator has been used. The aligned fiber exhibits high efficiency and the system has recorded spectra at up to 3 million "counts" per sec on strongly cathodoluminescent doped yttrium aluminium garnet (YAG) samples, at 30 kV and approximately 3 nA. A resolution of 3 nm [full width half maximum (FWHM)] at 621 nm has been measured from a red laser diode spectrum, on the ESEM column. A range of uncoated materials has been measured to characterize the CL system; these materials ranged from strongly luminescent YAG to weakly luminescent polymers. Well-characterized doped zircons have also been investigated. These data suggest that the previously reported intrinsic peak from zircon is a consequence of high-beam currents. PMID- 10675443 TI - Correlation between Charge Contrast Imaging and the Distribution of Some Trace Level Impurities in Gibbsite. AB - Charge contrast images (CCI) of synthetic gibbsite obtained on an environmental scanning electron microscope gives information on the crystallization process. Furthermore, X-ray mapping of the same grains shows that impurities are localized during the initial stages of growth and that the resulting composition images have features similar to these observed in CCI. This suggests a possible correlation between impurity distributions and the emission detected during CCI. X-ray line profiles, simulating the spatial distribution of impurities derived from the Monte Carlo program CASINO, have been compared with experimental line profiles and give an estimate of the localization. The model suggests that a main impurity, Ca, is depleted from the solution within approximately 3-4 um of growth. PMID- 10675444 TI - Quantitative Short-pulse Acoustic Microscopy and Application to Materials Characterization. AB - A new acoustic microscopy method was developed for providing near-surface elastic property mapping of a material. This method has a number of advantages over the traditional V(z) technique. First, it enables one to perform measurements in an automated mode that only requires user intervention in the setup phase. This automated mode makes it feasible to obtain quantitative microscopy images of the elastic property over an area on the material being tested. Also, it only requires a conventional ultrasonic system operating in pulsed mode for collecting the data, rather than a specialized tone-burst system, which is needed in the traditional quantitative scanning acoustic microscopy technique. Finally, unlike the traditional method, the new experimental process does not require calibration of the system's electronics or additional reference data taken under hard-to duplicate identical conditions from a material that does not exhibit surface acoustic waves. PMID- 10675446 TI - News and Commentary. PMID- 10675445 TI - Compression of Secondary Ion Microscopy Image Sets Using a Three-dimensional Wavelet Transformation. AB - This article proposes a lossy three-dimensional (3-D) image compression method for 3-D secondary ion microscopy (SIMS) image sets that uses a separable nonuniform 3-D wavelet transform. A typical 3-D SIMS measurement produces relatively large amounts of data which has to be reduced for archivation purposes. Although it is possible to compress an image set slice by slice, more efficient compression can be achieved by exploring the correlation between slices. Compared to different two-dimensional (2-D) image compression methods, compression ratios of the 3-D wavelet method are about four times higher at a comparable peak signal-to-noise ratio (PSNR). PMID- 10675447 TI - Diagnosis of lateral hypopharyngeal pouches: a comparative study of videofluorography and pseudovalsalva maneuver in double contrast pharyngography. AB - BACKGROUND: To evaluate the difference between the pseudovalsalva maneuver in double-contrast pharyngography and the videofluorographic swallowing examination in the detection and grading of lateral hypopharyngeal pouches. METHODS: Two hundred twenty-seven videofluorographic swallowing examinations and double contrast pharyngography using the pseudovalsalva maneuver were retrospectively analyzed by two radiologists. The mean age of the patients was 54 years (range = 21-81 years). The examination was performed on a fluoroscopy unit with a U-matic videorecording system in standard projections. Iodinated contrast agent was used, followed by barium if there was no massive aspiration. RESULTS: In contrast to the videofluorographic swallowing examination, which showed 170 lateral hypopharyngeal pouches (113 grade I, 39 grade II, 18 grade III) in 101 patients, the pseudovalsalva maneuver showed 304 pouches (304 grade III) in 179 patients. No videofluorographically diagnosed lateral hypopharyngeal pouches were missed by the pseudovalsalva maneuver; 134 pouches in 78 patients diagnosed with pseudovalsalva maneuver had no correlation videofluorographically. CONCLUSION: Double-contrast pharyngography using the pseudovalsalva maneuver is not a reliable method for the diagnosis of lateral hypopharyngeal pouches. PMID- 10675448 TI - Esophagobronchial fistula following redo Nissen fundoplication. AB - Gastrobronchial fistula is a rare complication of antireflux surgery, whereas esophagobronchial fistula as a complication of Nissen fundoplication has, to the best of our knowledge, not been reported previously. We report on a case of esophagobronchial fistula in a patient with left subphrenic abscess following redo Nissen fundoplication. Chest radiographs suggested an unresponsive pneumonia of the left lower lobe. Computed tomography (CT) of the abdomen showed partial consolidation of the left lower lobe and contrast filling of the left bronchial tree from a left subphrenic abscess. CT diagnosis of fistula originating from the region of fundoplication was confirmed by Gastrografin follow-through. PMID- 10675449 TI - Heterotopic pancreas of the stomach: CT findings correlated with pathologic findings in six patients. AB - BACKGROUND: The purpose of this study was to characterize the computed tomographic (CT) findings of heterotopic pancreas of the stomach. METHODS: CT scans of six surgically proven cases of heterotopic pancreas of the stomach were reviewed. Three were dynamic spiral CT scans, with both arterial dominant and late phase scans. In other three, both unenhanced and contrast-enhanced scans were obtained by using conventional techniques. Particular attention was given to the enhancement of the heterotopic pancreas. Pathologic and surgical findings were correlated with CT findings. RESULTS: The locations were in the gastric antrum in five cases and in the mid-body in one. Size ranged from 1 cm to 3 cm (mean = 2.1 cm). Three cases showed homogeneous, strong enhancement similar to the pancreas and consisted mainly of pancreatic acini with the same histologic features as the normal pancreas. Two cases showed poor enhancement and consisted mainly of ducts and hypertrophied muscle; pancreatic acini were a minor component. In one case appearing as a cystic lesion on CT, a pseudocyst was found with many ducts and some nests of pancreatic acini. CONCLUSIONS: Heterotopic pancreas of the stomach showed a diverse spectrum of CT findings. Good understanding of these CT findings may be helpful in making a correct diagnosis. PMID- 10675450 TI - Gastric adenoma with atypical appearance: findings on double-contrast barium study with histopathologic correlation. AB - BACKGROUND: To describe the radiologic findings of nonpolypoid gastric adenomas and to correlate them with pathologic findings. METHODS: During a 9-year period, we reviewed 49 pure gastric adenomas in 43 patients with positive radiologic findings. Of these adenomas, seven with atypical polypoid appearance were retrospectively included in the study. We reviewed these findings with double contrast barium study and correlated them with the pathologic findings. RESULTS: Of seven nonpolypoid adenomas, four were depressed and three were flat at pathologic examination. All were diagnosed as early gastric carcinoma (five as type IIc, one as type IIb, one as type IIa + IIc) in upper gastrointestinal series. Three were located in the gastric angle, two in the lower body, and two in the antrum. Size ranged from 10 mm to approximately 25 mm (mean = 15 mm). Six lesions had nodular surface and five had convergency of the mucosal folds. A shallow depressed area was seen in six lesions. CONCLUSIONS: A considerable proportion of gastric adenomas presents as a depressed or flat lesion on double contrast barium study because of histologic characteristics of decreased subjacent mucosa. Because the nonpolypoid adenoma has a greater potentional for malignancy, more precaution is needed during the follow-up of this uncommon lesion. PMID- 10675451 TI - Afferent loop syndrome presenting as enterolith after Billroth II subtotal gastrectomy: a case report. AB - We present a rare late-onset (after 24 years) complication of gastric surgery with a combination of afferent loop syndrome associated with a large duodenal stone. The patient, who had undergone Billroth II partial gastrectomy for benign ulcer 24 years before, developed abdominal pain in the right upper quadrant, associated with nausea, vomiting, and high grade fever. Abnormal laboratory values included elevated liver function test, suggesting a pressure-related phenomenon. Leukocytosis and a high level of platelets were also found. Only computed tomography and endoscopy of the upper gastrointestinal tract confirmed the diagnosis of a huge stone in the dilated duodenal afferent loop. To our knowledge, a case like this has not been reported previously in the literature. PMID- 10675452 TI - Spontaneously reduced midgut malrotation: CT diagnosis. AB - Midgut malrotation is a relatively rare congenital malformation that arises from an error of rotation and fixation of the midgut. We report a case of spontaneously reduced duodenal malrotation diagnosed by computed tomography and roentgenography after the ingestion of barium. PMID- 10675453 TI - Percutaneous balloon dilatation for benign hepaticojejunostomy strictures. AB - BACKGROUND: Percutaneous balloon dilatation of biliary tract strictures is generally accepted as a safe and inexpensive procedure. The effectiveness in selected groups of patients remains under discussion. The purpose of this study was to evaluate the results of percutaneous balloon dilatation in patients with a benign stricture of a hepaticojejunostomy. METHODS: Fifteen patients with a benign stricture of a hepaticojejunostomy were examined between 1993 and July 1997. An ultrasound-guided percutaneous transhepatic cholangiography (PTC) procedure was performed, followed by a balloon dilatation. Follow-up was performed prospectively by outpatient visits and laboratory testing. RESULTS: Percutaneous dilatation was successful in 14 patients. Three patients developed a recurrence. In one of these patients, the procedure was repeated successfully. Gastrointestinal bleeding occurred in one patient. The success rate for balloon dilatation in this group of patients was 73% after a mean follow-up of 30 months. When the procedure was repeated, the success rate was 80% after a mean follow-up of 33 months. CONCLUSIONS: Percutaneous balloon dilatation for benign hepaticojejunostomy strictures is feasible in the majority of patients and produces acceptable medium-term to long-term results. Advantages are its minimal invasive character and the fact that all options remain open in case of failure. PMID- 10675454 TI - Primary ileal plasmacytoma arising in mixed low- and high-grade B-cell lymphoma of mucosa-associated lymphoid tissue type. AB - We report an extremely rare case of primary ileal plasmacytoma accompanied by mixed low- and high-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type. The radiographic and macroscopic features of the tumor were characterized by two constricting lesions in the ileum. Histologic examination of the resected specimen showed that one constrictive lesion was plasmacytoma and the other MALT lymphoma was low and high grade. The plasmacytoma seemed to have differentiated from the MALT lymphoma. PMID- 10675455 TI - Mesenteric panniculitis: sonographic findings. AB - BACKGROUND: Mesenteric panniculitis (MP) is a relatively rare disease, and sonographic (US) and color Doppler findings have been infrequently reported. METHODS: We reviewed the clinical data and US and Doppler results of three cases of MP to determine the role and limitations of these techniques. RESULTS: The sole presenting clinical sign was a soft mass. On US the lesion was imaged as a poorly margined echogenic mass with hypoechoic areas. Color Doppler US visualized the nondeviated mesenteric vessels within the mass, which enabled us to perform a safe guided biopsy. CONCLUSIONS: US is useful as an initial diagnostic tool, but its results must be interpreted carefully. Color Doppler US is very useful in demonstrating fine vessels and helps in performing a safe needle biopsy. PMID- 10675456 TI - Altered flow dynamics of intravascular contrast material to the liver in superior vena cava syndrome: CT findings. AB - BACKGROUND: To evaluate the findings of altered flow dynamics in the livers of patients with obstruction of superior vena cava (SVC) on helical computed tomography (CT). METHODS: In six patients (age range = 28-80 years) with SVC obstruction, CT findings were retrospectively reviewed to identify the abnormal enhancement patterns of the liver and the relation with the extrahepatic collateral vessels and hepatic vessels. RESULTS: Abnormal hepatic enhancement was observed in the following four (A-D) portions: (A) anterior portion of segment IV (n = 5), (B) subdiaphragmatic portion of the liver (n = 4), (C) posterior portion of the right lobe (bare area; n = 1), and (D) lateral segment of the left lobe (n = 2). Two major collateral pathways to the liver were demonstrated as follows: A and D --> from the umbilical vein to the left portal vein, and B and C --> from the subcapsular vein to the bare area of the liver or to the hepatic veins. On helical CT, these collateral pathways were also clearly visualized. CONCLUSION: When these abnormal enhancements of the liver on CT are recognized within the liver, these findings indicate diversion of contrast material into collateral pathways to the liver with SVC obstruction. PMID- 10675457 TI - Detection of hepatocellular carcinoma and its metastases with various pulse sequences using superparamagnetic iron oxide (SHU-555-A). AB - BACKGROUND: To identify the most useful combinations of various pre- and postcontrast magnetic resonance (MR) image sequences in detecting hepatocellular carcinoma (HCC) and its intrahepatic metastases before and after injection of SHU 555-A. METHODS: Thirty-eight lesions in 16 patients were evaluated before and after administration of SHU-555-A by using fast spin echo (FSE), gradient echo (GRE), and echo planar (EP) imaging sequences using a 1.5-Tesla superconducting MR system. The signal intensity ratio (SIR) and contrast-to-noise ratio (CNR) of the lesions, signal-to-noise ratios, and other parameters were calculated. RESULTS: Tumors were better detected after injection of SHU-555-A on all pulse sequences except on out-of-phase T1-weighted (T1W)-GRE sequences. Tumor detectability was higher for precontrast EP imaging and T2*-weighted (T2*W)-GRE sequences, whereas detectability at postcontrast was higher for T2*W-GRE, proton density-weighted-FSE, and in-phase T1W-GRE sequences. The SIR and CNR at precontrast were highest for EP imaging, and those at postcontrast were highest for T2*W-GRE. CONCLUSION: SHU-555-A will increase the detectability of HCC and its liver metastases. T1W- and T2*W-GRE sequences would be the sequences of choice. PMID- 10675458 TI - Comparison of in-phase and out-of-phase gradient recalled echo T1-weighted pulse sequence for MR imaging of malignant liver masses following administration of paramagnetic gadolinium-chelate. AB - BACKGROUND: The purpose of this study was to compare the performance of in-phase and out-of-phase gradient recalled echo (GRE) pulse sequences on paramagnetic contrast-enhanced magnetic resonance (MR) imaging of malignant liver lesions. METHODS: Fifty patients (27 women, 23 men; mean age = 50 +/- 27 years) with known or suspected focal liver lesions, nine of whom had a fatty liver, were examined at 1.5 T before and 60 min after injection of gadobenate dimeglumine at a dose of 0.05 or 0.1 mmol/kg using two GRE techniques: echo time of 2.3 ms (out-of-phase) or 4.6 ms (in-phase). Liver signal-to-noise ratio (SNR) and lesion-liver contrast to-noise ratio (CNR) were calculated. RESULTS: In patients with a nonfatty liver, liver SNR increased from 26 +/- 9 to 41 +/- 17 on in-phase images and from 28 +/- 8 to 45 +/- 14 on out-of-phase images. In patients with a fatty liver, in-phase images provided significantly higher (p < 0.01) liver SNR than did out-of-phase images predose (34 +/- 8 on in-phase vs. 21 +/- 8 on out-of-phase) and postdose (44 +/- 13 on in-phase vs. 33 +/- 14 on out-of-phase). In patients with a nonfatty liver, lesion-liver CNR was similar on in-phase and out-of-phase images, predose and postdose. In patients with fatty liver, lesion-liver CNR was significantly (p < 0.01) lower on out-of-phase images on predose and postdose images. CONCLUSION: In-phase GRE imaging is recommended for imaging focal liver lesions on paramagnetic contrast-enhanced MR imaging in patients with fatty infiltration of the liver. PMID- 10675459 TI - Enhanced color flow images in small hepatocellular carcinoma. AB - BACKGROUND: Features of enhanced color flow images in small hepatocellular carcinoma (HCC) are not fully elucidated. The purpose of this study was to clarify the characteristic vascular images in small HCC observed by enhanced color Doppler. METHODS: Enhanced color Doppler using the contrast agent Levovist was performed on 13 patients with HCC smaller than 30 mm. Enhanced color flow appearance was compared with angiographic findings. Time-intensity changes after injection of the contrast agent were analyzed in HCC nodules. RESULTS: Significant improvement in the detection of color flow signals was obtained in small HCC using Levovist, from 33% in precontrast to 92% in postcontrast (p < 0.005). Three patterns of enhanced color flow images, which were related to the angiographic findings, were observed. The time-intensity curve was classified into two types by "time to peak" and "time on plateau" and was associated with the patterns of enhanced images. CONCLUSION: Enhanced color flow imaging promises to be a useful method for evaluating tumor vascularity noninvasively and to contribute to the elucidation of the hemodynamics in small HCC. PMID- 10675460 TI - Evaluation of the intratumoral vasculature of hepatocellular carcinoma by power doppler sonography: advantages and disadvantages versus conventional color doppler sonography. AB - BACKGROUND: To determine whether a difference exists in the relative ability of power Doppler sonography and conventional color Doppler sonography to detect the intratumoral vasculature of hepatocellular carcinoma based on lesion size and location. METHODS: Sixty patients with 88 hepatocellular carcinoma lesions that showed tumor staining on angiography and were enhanced on dynamic computed tomography were evaluated. Power Doppler sonography and color Doppler sonography were used to detect the intratumoral vasculature, and their sensitivity to blood flow was evaluated. RESULTS: Power Doppler sonography showed a superior detection rate for lesions smaller than 2 cm and located 4-8 cm from the abdominal surface in the right hepatic lobe as compared with color Doppler sonography (p < 0.01). Neither power Doppler sonography nor color Doppler sonography depicted the intratumoral vasculature of lesions located more than 8 cm from the abdominal surface (n = 14). Both color Doppler imagings exhibited a low detection rate for lesions in the left hepatic lobe (n = 31, p < 0.01). CONCLUSIONS: Power Doppler sonography should be applied in the evaluation of small or intermediate depth lesions because it is more sensitive to these lesions than color Doppler sonography, but it is not useful for left lobe and deep lesions. PMID- 10675461 TI - Intratumoral steatosis in focal nodular hyperplasia coinciding with diffuse hepatic steatosis: CT and MRI findings with histologic correlation. AB - Focal nodular hyperplasia (FNH) is a benign tumorlike condition that is thought to be a hyperplastic response to increased blood flow in an arterial malformation rather than a true neoplasm. Radiologically, FNH usually shows typical findings on unenhanced and enhanced computed tomography (CT) and magnetic resonance images (MRI), with atypical features being the exception rather than the rule. We report an unusual case of FNH with extensive fatty infiltration of the lesion illustrated on CT and MRI and proven by histopathology. PMID- 10675462 TI - Hepatic and splenic involvement in cat-scratch disease: imaging features. AB - Hepatosplenic involvement in cat-scratch disease, probably underdiagnosed, is characterized by multinodular lesions throughout the liver and spleen. Radiologic features of ultrasound, computed tomography, and magnetic resonance imaging are not specific. The key of the diagnosis relies on a history of cat or kitten contact. A specific serological test can confirm the diagnosis without invasive procedures such as biopsy. PMID- 10675463 TI - Massive intrahepatic extramedullary hematopoiesis in myelofibrosis. AB - We describe the sonographic (US) and computed tomographic appearances of a large, solitary tumor in the liver produced by extramedullary hematopoiesis in an 82 year-old patient with agnogenic myeloid metaplasia. Confirmation of this diagnosis was made by US-guided fine-needle aspiration biopsy. PMID- 10675464 TI - Intrahepatic splenosis: imaging features. AB - We report a patient who presented with asymptomatic focal liver lesions and in whom a diagnosis of intrahepatic splenosis was made. This rare condition mostly occurs in patients who previously underwent splenic trauma or surgery. Magnetic resonance imaging (MRI) characteristics suggesting this diagnosis are described. The lesions were mainly hypointense on T1- and hyperintense on T2-weighted images. After administration of small iron oxide particles (SPIO-Endorem), the lesions remained slightly hyperintense relative to the hypointense liver parenchyma but showed a 50% loss in signal intensity. Knowledge of these MRI characteristics may avoid the use of surgical interventions to arrive at the correct diagnosis of these rare liver lesions. PMID- 10675465 TI - Complications of "dropped" gallstones after laparoscopic cholecystectomy: technical considerations and imaging findings. AB - New laparoscopic techniques have revolutionized the practice of surgery. Laparoscopic cholecystectomy has become one of the most commonly performed surgeries worldwide. Although shorter hospital stays and patient comfort have offered clear advantages over open cholecystectomy, the technique has resulted in several specific complications, including bile duct injury and gallbladder perforation. Although rarely clinically significant, intraperitoneal gallstone spillage can cause abscess formation and adhesions. Although these patients can present with a confusing clinical picture, their characteristic radiologic features should be recognized. We present two cases of complicated intraperitoneal gallstone spillage radiologically diagnosed and treated with laparoscopic and interventional radiologic techniques. PMID- 10675466 TI - Gallbladder carcinoma: color Doppler sonography. AB - This study, based on color Doppler and pulsed Doppler sonographic results of 13 cases with gallbladder carcinoma, eight cases of adenomyomatosis, and eight cases of tumefactive biliary sludge, shows that the presence or absence of blood flow signals helps in the differentiation between gallbladder carcinoma and tumefactive biliary sludge (84.6% sensitivity and 80.0% specificity). However, color Doppler sonography is still not fully capable of distinguishing all gallbladder carcinoma, and a further increase in Doppler sensitivity is mandatory for this purpose. Visualization of high-velocity blood flow within the lesion made gallbladder carcinoma more likely than benign tumor. In contrast, there was no difference in the resistive index between gallbladder carcinoma, adenomyomatosis, and normal subject groups, and the significance of the resistive index is a subject of future study. PMID- 10675467 TI - Heterotopic gastric mucosa in the gallbladder: sonographic and CT findings. AB - We present computed tomographic (CT) and sonographic findings of heterotopic gastric mucosa incidentally found in a 63-year-old male. CT showed a slightly high density area in the gallbladder, which was intermediately enhanced early after bolus injection of contrast medium. Ultrasonography showed an echogenic sessile polyp. Histologically, the tumor consisted of gastric fundic glands containing parietal cells and chief cells. PMID- 10675468 TI - Serous and mucinous cystadenoma/cystadenocarcinoma of the pancreas. AB - Twenty cases of cystic pancreatic neoplasms were examined over a 10-year period by the Department of Radiology, University Hospital, Lund, Sweden. Four patients had serous cystadenoma, seven had mucinous cystadenoma, and seven had mucinous cystadenocarcinoma. One patient had a mucin-producing ductal carcinoma, and one patient had a benign mucus cyst. The various types of tumor are illustrated, and the difficulty in differentiating the subtypes is stressed. PMID- 10675469 TI - Hyaline vascular-type Castleman disease: a rare cause of a hypervascular retroperitoneal mass. AB - We present the cross-sectional imaging and angiographic findings of hyaline vascular-type Castleman disease located in the retroperitoneum. The diagnosis was made postoperatively. This entity can simulate a malignant neoplasm. The histologic subtypes and presentations of Castleman disease and the differential diagnosis of retroperitoneal masses are discussed. PMID- 10675470 TI - Paratesticular aggressive fibromatosis: CT findings. AB - Aggressive fibromatoses commonly originate from the musculoskeletal system, mesentery, and retroperitoneum. We report a case of aggressive fibromatosis arising from the spermatic cord. On helical computed tomography, the lesion appeared as a solid mass with well-defined borders in the scrotum and with infiltrative features in the retroperitoneum. PMID- 10675474 TI - Inhibition of malignant ascites and growth of human ovarian carcinoma by oral administration of a potent inhibitor of the vascular endothelial growth factor receptor tyrosine kinases. AB - We determined whether inhibition of the catalytic tyrosine kinase activity of the receptors for vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) inhibits the formation of malignant ascites and the progressive growth of human ovarian carcinoma cells implanted into the peritoneal cavity of nude mice. The novel protein tyrosine inhibitor PTK 787 was evaluated in two models of human ovarian cancer: Hey-A8 cells, which express low levels of VEGF/VPF and grow as solid tumor foci on the surface of peritoneal organs, and SKOV3 i.p.1 cells, which express high levels of VEGF/VPF and grow as solid peritoneal tumors and ascites. Treatment of nude mice by daily oral administration of 50 mg/kg PTK 787 was not effective against Hey-A8 tumors. In sharp contrast, it significantly inhibited growth of SKOV3 i.p.1 cells and formation of ascites, significantly increasing survival of mice with the implants. Tumor-induced vascular hyperpermeability in the peritoneum of tumor-bearing mice was inhibited by PTK 787, which accounted for its inhibition of ascites formation. Our results suggest that blockade of the VEGF/VPF receptor may be an efficient strategy to inhibit formation of malignant ascites and growth of VEGF/VPF-dependent human ovarian carcinomas. PMID- 10675475 TI - DF3 expression in human gallbladder carcinoma: significance for lymphatic invasion. AB - DF3 (MUC 1) is a member of a family of high molecular weight glycoproteins. Recent studies have demonstrated that DF3 is expressed in tumors of various human organs, and may function as an anti-adhesion molecule that inhibits cell-to-cell adhesion, inducing tumor metastasis. However, expression patterns of DF3 have not yet been established in human gallbladder carcinomas. In this study, we examined DF3 expression in human gallbladder adenocarcinoma and its clinicopathological significance. DF3 immunoreactivity was detected not only in the cancer cells (cytoplasmic type; 50.0%, 27/54) but also in the cancer stroma (stromal type; 46.3%, 25/54). According to TNM classification, 65.0% (26/40) of T2-4 gallbladder cancers showed cytoplasmic DF3, while 7.1% (1/14) of the T1 cancers were cytoplasmic DF3-positive (p<0.001). Stromal DF3 expression was detected in 62.5% (25/40) and none (0/14) of the T2-4 and T1 cancers, respectively (p<0.001). Lymph node metastasis was frequently found in the cytoplasmic DF3- and stromal DF3 positive gallbladder cancers (59.3% and 60.0%, respectively). These observations suggested that DF3 expression plays important roles in cancer cell growth and metastasis of human gallbladder adenocarcinomas. PMID- 10675476 TI - Microdissection based comparative genomic hybridization analysis (micro-CGH) of secondary acute myelogenous leukemias. AB - Comparative genomic hybridization (CGH) is a well established technique in molecular cytogenetics. However, leukemias, and especially secondary acute myelogenous leukemias (sAML) are not very well analyzed by this technique, even though such diseases are often characterized by complex karyotypic changes, not resolvable by conventional cytogenetic banding analysis. This lack of CGH-studies might be due to the fact, that in most cases bone marrow aspirate is too limited to do DNA-extraction additionally to the cytogenetic analysis. To circumvent this problem a new CGH technique has been applied to analyze 10 AML cases with complex karyotypic changes. In each case 15 interphase nuclei of the harvested and fixed bone marrow cell-suspension have been microdissected from the coverslip surface and collected in a tube. Subsequently, DNA was amplified by DOP-PCR. With this micro-CGH technique additional cytogenetic information from 10 highly aberrant AML cases was obtained and confirmed by FISH on metaphase of the corresponding AML case. PMID- 10675477 TI - Relationship between p53 gene mutation and protein expression: clinical significance in transitional cell carcinoma of the bladder. AB - Although the mutated p53 gene has been postulated to induce immunohistochemically detectable p53 protein, reports regarding the relationship between p53 mutation and p53 protein expression have been contradictory. This study investigated the relationship between p53 mutations and p53 expression and their clinical significance for patients with transitional cell carcinoma of the bladder. Eighty seven transitional cell carcinoma of the bladder were analyzed by immunohistochemistry (IHC) for p53 nuclear accumulation, and the results compared to mutations detected in the p53 gene evaluated by polymerase chain reaction single-strand conformation polymorphism (SSCP) and DNA sequence analysis. By p53 IHC analysis, positive p53 staining was observed in 50 (57.5%) of the 87 tumors. The specificity of IHC, defined as a percentage of IHC negative (<20%) tumors among tumors without mutation, was 94.6%. Despite the good concordance between p53 mutation and p53 protein expression (p<0.0001), 48.0% (24/50) of the tumors showed p53 overexpression without mutation, and 2 (5.4%) tumors with mutation showed no p53 immunoreactivity. Patients with higher grade (grade 3), stage (stages pT2-4), and p53 mutations had a poorer prognosis by Kaplan-Meier survival analysis. A Cox univariate analysis found that grading (hazard ratio 3.139; p=0.002), staging (hazard ratio 3.832; p=0.0005) and p53 mutation (hazard ratio 2.498; p=0.013) were significant variables in these patients, but no variable was independently associated with an increased survival of bladder carcinoma by multivariate analysis. We found that a 20% cut-off level of p53 overexpression showed the highest correlation with prognosis and p53 mutation, however, p53 overexpression and mutation were not superior to staging as prognostic markers. These data suggest that careful assessment of the TNM staging system remains the most reliable predictive indicator of survival for patients with transitional cell carcinoma of the bladder. PMID- 10675478 TI - Expression and mutation patterns of p53 in benign and malignant salivary gland tumors. AB - The expression and mutation patterns of p53 were studied in a series of 68 benign pleomorphic adenomas and 237 malignant salivary gland tumors. p53 overexpression (nuclear staining exceeding 10%) was detected in 20% of the malignant salivary gland tumors, with the highest prevalence observed in polymorphous low grade adenocarcinoma, squamous cell carcinoma, and carcinoma ex pleomorphic adenoma and the lowest in adenoid cystic carcinoma and acinic cell carcinoma. In contrast, none of the 68 benign pleomorphic adenomas had nuclear staining exceeding 10%. SSCP and nucleotide sequence analysis of exons 4 to 9 of p53 in 19 malignant tumors revealed 9 mutations in 7 tumors. Our findings indicate that p53 may be a useful marker to help discriminate between benign and malignant salivary gland tumors. PMID- 10675479 TI - Identification of a novel subtype of H4-RET rearrangement in a thyroid papillary carcinoma and lymph node metastasis. AB - RET/PTC chimeric oncogenes are generated by the fusion of heterologous genes to the RET tyrosine kinase encoding domain. These rearrangements are typical of papillary thyroid carcinomas. RET/PTC1 is one of the most frequently found RET/PTC version and, in all the cases so far reported, it is invariably generated by the fusion of the first encoding exon of the H4 gene to the RET kinase encoding domain. This results in the generation of an oncogenic protein containing the first 101 residues of the H4 protein at the N-terminus. We report the isolation of a novel subtype of H4-RET fusion, designated RET/PTC1L, from a human papillary carcinoma of the thyroid and lymph node metastasis. At variance with the classic one, this novel rearrangement generates a protein containing the N-terminal 150 residues of H4. RET/PTC1L is able to transform NIH 3T3 cells; its transforming ability, however, is 5-fold lower than that of the classic RET/PTC1 isoform. We propose that RET/PTC1L is a novel chimeric oncogene involved in thyroid tumorigenesis; its low transforming ability may be one of the reasons explaining the low frequency by which it is found in human thyroid carcinomas. PMID- 10675480 TI - Missense mutation of the hMSH6 and p53 genes in sporadic urothelial transitional cell carcinoma. AB - Functional defects in DNA mismatch repair genes have been shown to be associated mainly with hereditary human malignancies. We examined genomic DNA from 88 sporadic transitional cell carcinomas (TCCs) of the urinary tract for mutations in hMSH6 gene by polymerase chain reaction and direct sequencing analysis. Mutational status of p53 gene was also studied as a potential target of genetic instability secondary to hMSH6 dysfunction. A total of 5 cases (5.7%: 5/88) displayed hMSH6 mutations all consisted of transition and located in exon 4, including three cases with missense mutation and two without change of corresponding amino acid. These three tumors with hMSH6 missense mutation had no microsatellite instability with five microsatellite markers tested. p53 gene mutations were detected in 22 cases (25.0%: 22/88). No tumors with p53 mutation had any hMSH6 missense mutations. Compared to the cases without hMSH6 alterations, the three patients with hMSH6 alterations had more frequent additional primary cancer (P<0.05). These findings provide the first in vivo evidence for the type of alterations and frequency of possible involvement of the hMSH6 mutations in sporadic type urothelial TCCs. PMID- 10675481 TI - Regulation of Bcl2 phosphorylation by stress response kinase pathway. AB - The anticancer drugs affecting either microtubule polymerization or depolymerization could trigger Bcl2 phosphorylation in mitotic phase of the cell cycle. By systematic site directed mutagenesis studies, we have previously mapped taxol induced phosphorylation sites to be Ser-70 and 87 residues of Bcl2 protein. Interestingly, sequences surrounding both serine-70 and serine-87 residues represent MAP kinase consensus motif. Since Bcl2 phosphorylation predominantly occurs at site consensus to MAP kinase family members, we were interested to test whether Erk2 or Jun kinase is involved in this pathway. Our in vitro studies document that stress activated kinase, JNK1 is responsible for Bcl2 phosphorylation. PMID- 10675482 TI - The K-ras gene regulates vascular endothelial growth factor gene expression in non-small cell lung cancers. AB - Tumor angiogenesis is an essential step for tumor cell growth, progression and metastasis. Vascular endothelial growth factor (VEGF) is mitogen specific for endothelial cells, and therefore is believed to play a key role in tumor angiogenesis. However, the mechanisms underlying the regulation of VEGF expression remain virtually unknown and the only major regulator of VEGF expression has been reported to be hypoxia. Recently, it was reported that a mutant p53 in#duced the expression of VEGF mRNA, and that wild-type p53 down regulated endogenous VEGF mRNA levels. In contrast, it has also been reported that mutant ras oncogenes were associated with the marked up-regulation of VEGF in transformed epithelial cells. Based on these results, we performed a retrospective study of the p53 and K-ras genes status and VEGF gene expression in the tumor tissues from 181 patients with non-small cell lung cancer using SSCP, sequencing, RT-PCR and immunohistochemical techniques. Forty-six carcinomas (25.4%) were evaluated as having high VEGF expression, and 135 tumors (74.6%) had low VEGF expression. Of the 181 primary NSCLC studied, 63 carcinomas (34.8%) contained mutations of p53, whereas only 14 carcinomas (7.7%) had mutations of K ras. There were no significant relationships between VEGF expression and p53 status or each mutant exon of p53. In contrast, a significant difference was found between VEGF expression and K-ras status. Of the 14 tumors with mutant K ras genes, 7 cases (50.0%) had high VEGF expression whereas only 39 of the 167 tumors with wild-type K-ras (23.4%) had high VEGF expression (p=0.0278). The mean VEGF conservation rate for the 14 tumors with mutant K-ras genes was 0.77+/-0.58 and the rate of the 167 tumors with wild-type K-ras genes was 0.49+/-0.46 (p=0. 0350). Moreover, the overall survival rate of patients with high VEGF expression was lower than patients with low VEGF expression (45.7% vs 60.7%, p=0.0419). On the other hand, there was no significant difference in the overall survival rate between patients with a mutant p53 and those with a wild-type p53; there was also no difference in the overall survival between patients with a mutant K-ras and those with a wild-type K-ras. The Cox regression model analysis indicated that three variables, VEGF status, K-ras status and nodal status, were found to be significant indicators for prognosis (p=0.0236, p=0.0172 and p<0.0001, respectively). Our data suggest that a high expression of VEGF in lung cancer may be associated with a poor prognosis. This may be a clue to improving lung cancer diagnoses and therapies aimed at inhibiting tumor angiogenesis due to VEGF. PMID- 10675483 TI - Activation of voltage-operated Ca2+-channels in human small cell lung carcinoma by the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone. AB - The nicotine-derived tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) induces lung cancer in all animal species tested and is thought to contribute significantly to the high lung cancer burden associated with smoking. NNK has recently been identified as a high affinity ligand for neuronal nicotinic acetylcholine receptors comprised of alpha7 subunits (alpha7 nAChR), and expressed in human small cell lung carcinoma (SCLC). As agonist binding to this receptor in mammalian cells often results in membrane depolarization and activation of voltage-operated Ca2+-channels (VOCCs), we hypothesized that NNK may exert similar effects in SCLC. Using flow cytometry to monitor the influx of Ca2+, reverse transcription polymerase chain reaction (RT PCR) to determine the expression of VOCC-specific messenger RNA, as well as analysis of DNA synthesis or determination of cell number, our data demonstrate that binding of NNK to the alpha7 nAChR in SCLC cells caused influx of Ca2+ via VOCCs of the L-, N-, and P-type. In turn, this led to a significant increase in DNA synthesis and cell number which was inhibited by a site-selective antagonist for the alpha7 nAChR and by Ca2+-channel blockers of the L-, N-, or P-types of VOCCs. Our findings suggest that the chronic activation of VOCC-mediated Ca2+ influx by NNK in smokers is an important event that may affect numerous Ca2+ dependent signal transduction pathways, thus contributing significantly to the development of SCLC. PMID- 10675484 TI - Mechanisms in the chemoprevention of colon cancer: modulation of protein kinase C, tyrosine protein kinase and diacylglycerol kinase activities by 1,4 phenylenebis-(methylene)selenocyanate and impact of low-fat diet. AB - Epidemiological and experimental studies suggest an inverse relationship between the intake of dietary selenium and/or low fat-intake and colon cancer risk. Efficacy studies in rodents suggest that the organoselenium compound 1, 4 phenylenebis(methylene)selenocyanate (p-XSC), is a more effective and less toxic chemopreventive agent than other organic or inorganic selenium compounds such as selenomethionine and Na2SeO3. The efficacy of p-XSC against colon cancer is significantly augmented by a low-fat diet. To explore the mechanisms by which this combined inhibiting effect against colon carcinogenesis comes about, we have investigated protein kinase C (PKC), tyrosine protein kinase (TPK), diacylglycerol kinase (DGK) activities and 8-isoprostane levels in colonic mucosa and tumor tissues in an azoxymethane (AOM)-induced rat colon cancer model. Weanling male F344 rats were fed the semipurified AIN-76A diet until seven weeks of age. Then various experimental groups were fed the low- or high-fat diets containing 0 or 20 ppm p-XSC (10 ppm as selenium). At seven weeks of age, groups of rats were injected s.c. with azoxymethane (AOM; 15 mg/kg body wt., once weekly for 2 weeks) and continued on their respective experimental diets until 38 weeks after the second AOM treatment. They were then sacrificed and colonic mucosal and tumor samples were evaluated for PKC, TPK, DGK and 8-isoprostane levels. Administration of p-XSC along with a low-fat diet significantly inhibited Ca+2 dependent and -independent PKC (P<0.05-0.01) activities in colonic mucosa and tumors. Administration of p-XSC either low-fat or high-fat diet significantly suppressed both colonic mucosal and tumor TPK activity (P<0.05-0.01). Suppression of TPK activity was more pronounced in rats maintained on a low-fat diet containing p-XSC. In contrast, rats receiving p-XSC with either low- or high fat diet showed significantly increased DGK activity (P<0.01-0.0001). Rats fed low fat or high-fat plus p-XSC had lower-levels of 8-isoprostane in the colonic tumors than animals who had been given low- or high-fat diets without the organoselenium compound. Interestingly, 8-isoprostane levels were lower in the colon tumors of the rats fed the low-fat diet than those fed the high-fat diet. Our findings suggest that p-XSC induced down-regulation of PKC and TPK activities and up-regulation of DGK activity. These events may in part be responsible for the chemopreventive activity against colon carcinogenesis. Further, this study implies that p-XSC with a low-fat dietary regimen will augment regulation of PKC, TPK and DGK activities in the colon. PMID- 10675485 TI - Association of renal cell carcinoma antigen, disialylgalactosylgloboside, with c Src and Rho A in clustered domains at the surface membrane. AB - Disialylgalactosylgloboside (DSGG), defined by monoclonal antibody RM2, is a renal cell carcinoma (RCC)-associated antigen which mediates adhesion of RCC TOS 1 cells to certain lung tissue target cells. This adhesion process may initiate preferential lung metastasis of RCC. Ganglioside GM3 is a B16 melanoma-associated antigen which similarly adheres to target cells and promotes consequent metastasis. In view of the close association of GM3-enriched microdomain with transducer molecules c-Src, Rho A, and FAK in B16 cells, we investigated the organizational status of DSGG in RCC cell line TOS-1, with the following results: i) DSGG, but not monosialylgalactosylgloboside, showed extensive clustering at the TOS-1 cell surface; ii) a low-density membrane fraction isolated from TOS-1 cells contained >95% of cellular DSGG, although protein content in this fraction was <1% of total cellular protein; iii) this fraction contained c-Src, Rho A, and FAK, but not H-Ras; iv) c-Src and Rho A were co-immunoprecipitated with DSGG through anti-DSGG mAb RM2 (IgM) affixed to a column. These observations indicate that DSGG is clustered in RCC, as typified by TOS-1 cells, to form a microdomain in which it is closely associated with c-Src, Rho A, and FAK, and may constitute a functional unit as has been observed for GM3 with transducer molecules in B16 cells. The functional organization of such units may be essential in determining malignant properties of RCC cells. PMID- 10675486 TI - Alteration of the CDKN2/p16 gene is not required for HPV-positive uterine cervical cancer cell lines. AB - To determine whether alterations of the CDKN2/p16 might be involved in HPV positive cervical cancers, we examined for alterations of this gene and function of the protein p16 to interact with CDK4 in 5 cervical cancer cell lines. No alteration of this gene was detected. Proteins for p16 and CDK4 were normally expressed and function of p16 to interact with CDK4 was not abrogated in these cell lines. These cell lines were human papillomavirus (HPV)-positive and carried wild-type p53. These findings suggest that phosphorylation of pRb by CDK4 is not critical in the carcinogenesis or in the establishment of HPV-positive cervical cancer cell lines, since HPV E6 or E7 viral-transforming proteins inactivate p53 and pRb tumor suppressor protein function, resulting in deregulated progression of the cell cycle. PMID- 10675487 TI - Infrequent widespread microsatellite instability in hepatocellular carcinomas. AB - Widespread or high-frequency microsatellite instability (MSI) due to the defective DNA mismatch repair (MMR) occurs in the majority of hereditary non polyposis colorectal cancer and a subset of sporadic malignant tumors. The incidence of MSI and underlying DNA MMR defects have been well characterized in gastrointestinal carcinogenesis, but not in hepatocarcinogenesis. To address the issue, we analyzed 55 Japanese hepatocellular carcinomas using several indicators of DNA MMR defects, such as microsatellite analysis, loss of heterozygosity (LOH) and mutation analysis of MMR genes, methylation of hMLH1 promoter, and frameshift mutations of mononucleotide repeat sequences within possible target genes. Mutation of beta2-microglobulin gene, which is presumably involved in MSI positive tumor cell escape from immune surveillance was also examined. Some of these analyses were also carried out in 9 human liver cancer cell lines. None of the 3 quasi-monomorphic mononucleotide markers sensitive for MSI, BAT26, BAT25, and BAT34C4 presented shortened unstable alleles in any of the carcinoma, cirrhosis, chronic hepatitis tissues, or cell lines. LOH at MMR genes was infrequent (4.4 approximately 7.1%), and no mutations were detected. Neither hMLH1 hypermethylation nor frameshift mutation in the target genes was detected. No mutations were found in beta2-microglobulin. Widespread MSI due to the defective DNA MMR appears to play little if any part in Japanese hepatocarcinogenesis. PMID- 10675488 TI - Analysis of glycoproteins in cancers and normal tissues reactive with monoclonal antibodies B3 and B1. AB - In this study we show by immunoblotting that B1 and B3, two newly isolated monoclonal antibodies, react with a variety of glycoproteins with different molecular weights expressed in stomach, pancreas, colorectal and breast cancers. The pattern of reactivity differed among cancers arising in different tissues, although no correlation has been observed with the histopathological characteristics of the lesion analysed. MAb B3 and MAb B1, have a limited reactivity with peritumoral tissues, whereas react very strongly with metastatic lesion. Because of the limited reactivity of these antibodies with normal tissue, MAbs B3 and B1, armed with toxin in the form of recombinant immunotoxins, can be useful in treating certain kinds of cancer such as metastatic lesions. However, until current clinical trials are completed, we will not know if they will be helpful in cancer treatment. PMID- 10675489 TI - Comparison of two human ovarian carcinoma cell lines (A2780/CP70 and MCAS) that are equally resistant to platinum, but differ at codon 118 of the ERCC1 gene. AB - ERCC1 is an essential gene within the nucleotide excision repair process. We studied two human ovarian carcinoma cell lines for cisplatin resistance, which differed with respect to ERCC1. The A2780/CP70 cell line has been extensively studied previously, and has the wild-type ERCC1 sequence. The MCAS cell line has a recently described ERCC1 polymorphism at codon 118, which is associated with an approximate 50% reduction in codon usage. These cells did not differ with respect to p53 sequence nor p53 mRNA induction following cisplatin exposure. The induction of ERCC1 mRNA was markedly reduced in MCAS cells as compared to A2780/CP70 cells. At the IC50 cisplatin dose for each cell line, MCAS cells were less proficient at cisplatin-DNA adduct repair than A2780/CP70 cells. In absolute terms, A2780/CP70 cells repaired 3-fold as much adduct (2.7 pg/microgram DNA over 6 h vs 0.86 pg/microgram DNA); and when expressed in terms of the maximal DNA adduct load, A2780/CP70 cells repaired 50% more adduct than MCAS cells. MCAS cells had increased cytosolic inactivation of drug at the IC50 dose level, which has been previously suggested to be a compensatory cellular response for reduced DNA repair capacity. These data suggest the possibility that this specific ERCC1 polymorphism, may be associated with reduced DNA repair capacity in human ovarian cancer cells. This association may be effected through a reduction in peak production of ERCC1 mRNA, and a consequent reduction in the translation of ERCC1 mRNA into protein. PMID- 10675490 TI - Downregulation of the p53 tumor suppressor gene and upregulation of the bcl-2 gene in retinoic acid receptor alpha-deficient transgenic mice. AB - We recently demonstrated lymphoma development in transgenic mice deficient in retinoic acid receptor alpha (RARalpha). High incidence of lymphoma development in this transgenic mouse model system was similar to lymphoma development in p53 knockout mice. In an effort to understand the molecular basis of lymphomagenesis in RARalpha-deficient transgenic mice, we compared the levels of RARalpha to the levels of p53 mRNA, and Bcl-2, and Bax proteins in lymphoid and non-lymphoid tissues and in lymphomas derived from the RARalpha-deficient transgenic mice. The p53 mRNA levels were depleted in various tissues including spleen ( approximately 96%), thymus ( approximately 29%) and bone marrow ( approximately 62%) of RARalpha-deficient transgenic mice when compared with the normal littermates, and the reduction in p53 mRNA expression in the various tissues examined was proportional to the reduction in RARalpha expression. Bcl-2 to Bax ratios were highly increased in the lymphoid compartments (spleen >bone marrow >thymus) because of selective overexpression of Bcl-2 protein. In summary, RARalpha downmodulation in this transgenic mouse model system was accompanied by p53 downmodulation and deregulation of Bcl-2 to Bax ratios in the lymphoid compartments. PMID- 10675491 TI - Major oncogenes and tumor suppressor genes involved in epithelial ovarian cancer (review). AB - Ovarian cancer remains the leading cause of death from gynecologic malignancy in Western countries. This cancer results from a succession of genetic alterations involving oncogenes and tumor suppressor genes which have a critical role in normal cell growth regulation. Mutations and/or overexpression of three oncogenes, HER-2/neu, c-myc and K-ras, and of the tumor suppressor gene p53, have frequently been observed in sporadic ovarian cancer. In the context of high risk families, the most frequently involved genes are BRCA1 and BRCA2. We review the function of these different proteins, the incidence of mutations in their genes in carcinogenesis and as potential prognostic factors in sporadic and hereditary ovarian cancer. PMID- 10675492 TI - Autocrine secreted insulin-like growth factor-I stimulates MAP kinase-dependent mitogenic effects in human primitive neuroectodermal tumor/medulloblastoma. AB - Primitive neuroectodermal tumors/medulloblastoma (PNET/MB) have similarities to neuroectodermal progenitor cells of the developing CNS. Since insulin-like growth factor I (IGF-I) exerts pleiotrophic effects on cells in the developing CNS, we evaluated the production, mitogenic effects and signaling pathways of IGF-I in PNET/MB cells and found that IGF-I is an autocrine growth factor in human PNET/MB cell lines tested. Stimulation of DAOY cells by IGF-I led to phosphorylation of its cognate receptor (IGF-IR) and resulted in cell proliferation. These effects of IGF-I were suppressed by IGF-IR blocking antibodies and by PD 98059, MAP kinase pathway inhibitor. The results demonstrate the existence of an autocrine IGF-I/IGF-IR loop and indicate that IGF-I promotes proliferation via MAP kinase pathway. PMID- 10675493 TI - The p16INK4alpha/p19ARF gene mutations are infrequent and are mutually exclusive to p53 mutations in Indian oral squamous cell carcinomas. AB - Eighty-seven untreated primary oral squamous cell carcinomas (SCCs) associated with betel quid and tobacco chewing from Indian patients were analysed for the presence of mutations in the commonly shared exon 2 of p16INK4alpha/p19ARF genes. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing analysis were used to detect mutations. SSCP analysis indicated that only 9% (8/87) of the tumours had mutation in p16INK4alpha/p19ARF genes. Seventy two tumours studied here were previously analysed for p53 mutations and 21% (15/72) of them were found to have mutations in p53 gene. Only one tumour was found to have mutation at both p53 and p16INK4alpha/p19ARF genes. Thus, the mutation rates observed were 21% for p53, 9% for p16INK4alpha/p19ARF, and 1% for both. Sequencing analysis revealed two types of mutations; i) G to C (GCAG to CCAG) transversion type mutation at intron 1-exon 2 splice junction and ii) another C to T transition type mutation resulting in CGA to TGA changing arginine to a termination codon at p16INK4alpha gene codon 80 and the same mutation will alter codon 94 of p19ARF gene from CCG to CTG (proline to leucine). These results suggest that p16INK4alpha/p19ARF mutations are less frequent than p53 mutations in Indian oral SCCs. The p53 and p16INK4alpha/p19ARF mutational events are independent and are mutually exclusive suggesting that mutational inactivation of either p53 or p16INK4alpha/p19ARF may alleviate the need for the inactivation of the other gene. PMID- 10675494 TI - Both HPV and carcinogen contribute to the development of resistance to apoptosis during oral carcinogenesis. AB - Oral carcinomas frequently contain human papilloma virus (HPV)-16/18. As p53 is degraded through interaction with HPV-16/18 products (E6/E7), p53 dysfunction may contribute to oral carcinogenesis. Furthermore, epidemiological studies suggest that smoking history may be critical for oral carcinogenesis. To delineate the involvement of HPV-16 infection and carcinogen in oral carcinogenesis, Park et al have established a multistep oral carcinogenesis model. Overexpression of p53 altered the expression of Fas antigen (Fas-R), Bax and Bcl-2; however, it remains unclear how the loss of p53 modifies the expression of these molecules. Using the multistep oral carcinogenesis model, we analyzed how the loss of p53 and carcinogen modified the expression of these molecules and their role in the development of resistance to apoptosis of oral carcinomas. The HOK-16B cell line was immortalized by HPV-16 transfection of normal human oral keratinocytes (NHOK). HOK-16B-BaP and HOK-16B-BaP-T1 were established from HOK-16B following short-term and long-term stimulation with the chemical carcinogen, benzo(a)pyrene, respectively. The malignant phenotype develops in sequence from HOK-16B, HOK-16B-BaP and HOK-16B-BaP-T1. The expression of apoptosis-related molecules was examined by Western blot analysis or by flow cytometry. Fas mediated cytotoxicity was assessed using CH-11, an agonistic anti-Fas-R IgM monoclonal antibody. The apoptosis-related molecules examined were the Fas-R, Bcl 2, Bax, and Fas-associated phosphatase 1 (FAP-1). Downregulation of Fas-R and upregulation of Bcl-2 in HOK-16B-BaP were observed in HOK-16B-BaP and HOK-16B BaPT1. Bax was downregulated in HOK-16B, HOK-16B-BaP and HOK-16B-BaP-T1. The expression of FAP-1 was increased with progression towards malignancy. NHOK and HOK-16B were relatively sensitive to CH-11, whereas HOK-BaP and HOK-BaP-T1 were resistant to CH-11. Treatment of HOK-16B-BaP with antisense bcl-2 oligonucleotide rendered the cells more sensitive to CH-11-induced apoptosis. These data demonstrate that both the loss of p53 and carcinogen stimulation are associated with altered expression of Fas-R, Bcl-2 and FAP-1, although the loss of p53 is sufficient for altered expression of Bax. Thus, both HPV infection and smoking contribute to acquisition of anti-apoptotic characteristics by oral carcinomas. PMID- 10675495 TI - Transforming growth factor-beta and response to anticancer therapies in human liver and gastric tumors in vitro and in vivo. AB - Liver cancer and gastric cancer are the most common solid tumors worldwide. Transforming growth factor-beta (TGF-beta) production and lack of response to TGF beta growth inhibitory effects have been associated with tumor progression and therapeutic resistance. HepG2, Hep3B, and SK-HEP-1 human liver cancer lines produce 3, 5.7, and 2.5 ng TGF-beta1; 1.4, 2, and 4 ng TGF-beta2 and 0.15, 0.2 and 0.22 ng TGF-beta3 per 107 cells (24 h). Expression of the TGF-beta type I receptor is 20x, 1x, and 0.6x the level in mink lung MvLu1 cells in the HepG2, Hep3B, and SK-HEP-1 cells, respectively. HepG2 and Hep3B cells do not express the TGF-beta type II receptor while SK-HEP-1 cells express 7x the level found in mink lung MvLu1 cells. Hs 746T, KATO III, RF-1, and RF-48 human gastric cancer cell lines produce 12. 5, 0.35, 0.4, and 0.4 ng TGF-beta1; 2.6, 0.95, 0.5, and 0.52 ng TGF-beta2 and 0.42, 0.17, 0.12, and 0.14 ng TGF-beta3 per 107 cells (24 h). Expression of TGF-beta type I receptor is 0.7x, 0.7x, 0.8x, 0.6x the level in mink lung MvLu1 cells in the Hs 746T, KATO III, RF-1 and RF-48 cells, respectively. KATO III cells are lacking in the TGF-beta type II receptor while Hs 746T, RF-1 and RF-48 cells express 10x, 0.8x, and 1x the levels in mink lung MvLu1 cells. The IC50 for TGF-beta1 is >>10 ng/ml in all of these lines except RF 48 where TGF-beta1 is mitogenic. The response of the cell lines to radiation, doxorubicin, mitomycin C, cisplatin, 5-fluorouracil, methotrexate, and gemcitabine showed that SK-HEP-1 was the most drug resistant liver cancer cell line and KATO III was the most drug resistant gastric cancer cell line. Overall, there was no correlation between TGF-beta secretion in cell culture and sensitivity of the cells to anticancer agents. Increased TGF-beta1 levels were detectable in the plasma of nude mice bearing Hep3B and Hs 746T xenografts. Those tumors which secreted greater amounts of TGF-beta were more therapeutically resistant in vivo. PMID- 10675496 TI - Radiotherapy for carcinoma of the posterior pharyngeal wall. AB - Posterior pharyngeal carcinoma has an extremely poor prognosis regardless of the method of treatment. The purpose of this study was to assess the local control and survival in patients with carcinoma of the posterior pharyngeal wall treated with definitive radiotherapy and to determine prognostic factors which may be relevant to the current UICC staging classification. Between January 1991 and December 1995, 22 patients with a mean age of 60 years (range 44-82) received definitive radiotherapy, using a homogeneous technique, for carcinoma of the posterior pharyngeal wall. The median follow-up was 42 months (range 25-66). The overall 3-year survival and local control for the whole group was 50% and 73% respectively. Patients with early stage (T1 and T2) disease had a significantly better overall 3-year survival rate of 77% compared to 11% for patients with advanced stage (T3 and T4) disease (p=0.0010). Similarly, patients with early stage disease had a significantly improved 3-year local control rate compared to patients with more advanced stage disease (92% and 44% respectively, p=0.0080). Patients with node positive disease had an inferior survival rate of 29% compared to 60% for those with node negative disease though the difference did not reach statistical significance. In addition only one patient with initial node negative disease had isolated nodal relapse. There was no significant late morbidity. For patients with early stage disease we have obtained local control and survival rates comparable to other groups with a once daily, short fractionation radiotherapy scheme but with reduced morbidity. In late stage disease altered fractionation schemes should be considered in order to achieve better local control and survival. Isolated nodal relapse was not a significant problem in this cohort of patients. Outcome correlates with primary tumour size and this is reflected in the current UICC staging classification. PMID- 10675497 TI - Mechanical deformation induces proliferation of human colorectal carcinoma cells. AB - The cell biology of intravascular tumor cells is clinically important but the many important variables of this environment have proved difficult to model. We studied the effects of repetitive mechanical deformation, a phenomenon affecting all intravascular cells, on human colon cancer cell line HCT 116 in vitro. Cell proliferation, assessed by [3H]-thymidine incorporation and cell count, increased by about 30% at two days in cells subjected to deformation at 30 cycles/min as compared to controls; levels of the nuclear proliferation antigen detected by monoclonal antibody MIB-1 were also increased. Deformation increased transforming growth factor beta1 (TGF-beta1) and plasminogen activator inhibitor-1 gene expression sevenfold at two days, but mannose-6-phosphate did not affect cell proliferation, indicating that endogenous TGF-beta is not involved in the proliferative response. HCT 116 cells lack TGF-beta type II receptors, but stable transfection of TGF-beta type II receptor cDNA did not alter the cellular response to mechanical deformation, as assessed by cell proliferation, morphology, or gene expression. Mechanical deformation affects several important aspects of HCT 116 cell biology, suggesting that the intravascular environment may regulate tumor cell biology in general. Endogenous TGF-beta and TGF-beta receptor-mediated signaling are not responsible for the deformation-induced proliferative response in HCT 116. PMID- 10675498 TI - Detection of telomerase activity in human carcinomas using a trap-ELISA method: correlation with hTR and hTERT expression. AB - We have evaluated telomerase activity in a tumour population of 65 human cancers by using a TRAP-based method, in which detection is performed by an enzyme immunoassay (ELISA). We have corroborated that sensitivity and specificity of this new procedure can be considered similar to that of classical TRAP method, having the advantage of a rapid and reproducible analysis of large pools of samples. Thus, telomerase activity was detected in 83% of the tumours included in our population. Moreover, we found a significant association between enzyme activity and both hTR and hTERT expression (P=0.004 and P=0.04, respectively). PMID- 10675499 TI - Mechanism of hepatocellular uptake of albumin-bound bilirubin. AB - We previously demonstrated that unconjugated bilirubin spontaneously diffuses through phospholipid bilayers at a rate which exceeds albumin dissociation, suggesting that solvation from albumin represents the rate-limiting step in hepatic bilirubin clearance. To further examine this hypothesis, we studied the uptake of bovine serum albumin (BSA)-bound bilirubin by cultured hepatoblastoma (HepG2) cells. Uptake of bilirubin was saturable, with a K(m) and V(max) of 4.2+/ 0.5 microM (+/-S.E.M.) and 469+/-41 pmol min(-1) mg(-1) at 25 degrees C. Substantial bilirubin uptake also was observed at 4 degrees C (K(m)=7.0+/-0.8 microM, V(max)=282+/-26 pmol min(-1) mg(-1)), supporting a diffusional transport mechanism. Consistent with reported solvation rates, the cellular uptake of bilirubin bound to human serum albumin was more rapid than for BSA-bound bilirubin, indicative of dissociation-limited uptake. Counterintuitively, an inverse correlation between pH and the rate of bilirubin flip-flop was observed, due to pH effects on the rate of dissociation of bilirubin from albumin and from the membrane bilayer. The identification of an inflection point at pH 8.1 is indicative of a pK(a) value for bilirubin in this range. Taken together, our data suggest that hepatocellular uptake of bilirubin is dissociation-limited and occurs principally by a mechanism involving spontaneous transmembrane diffusion. PMID- 10675500 TI - Effects of the hinge region of cecropin A(1-8)-magainin 2(1-12), a synthetic antimicrobial peptide, on liposomes, bacterial and tumor cells. AB - A 20-residue hybrid peptide (CA(1-8)-MA(1-12): KWKLFKKIGIGKFLHSAKKF-NH(2)) incorporating 1-8 residues of cecropin A (CA) and 1-12 residues of magainin 2 (MA) has potent antibiotic activity without hemolytic activity. In order to investigate the effects of the flexible hinge sequence, Gly-Ile-Gly of CA(1-8) MA(1-12) (CA-MA) on antibiotic activity, CA-MA and its three analogues, CA-MA1, CA-MA2 and CA-MA3 were synthesized. The Gly-Ile-Gly sequence of CA-MA was deleted in CA-MA1 and replaced with Pro and Gly-Pro-Gly in CA-MA2 and CA-MA3, respectively. CA-MA1 and CA-MA3 caused a significant decrease in the bactericidal rate against Escherichia coli and Bacillus subtilis and the tumoricidal activity against four different tumor cells, and the PC/PS (4:1, w/w) vesicle-aggregating and disrupting activities. However, CA-MA2 showed a similar bactericidal rate and antitumor, vesicle-aggregating and disrupting activities, as compared with CA-MA. These results suggested that the flexibility or beta-turn induced by Gly-Ile-Gly or Pro in the central part of CA-MA may be important in the electrostatic interaction of the cationic short alpha-helical region in the N-terminus with the cell membrane surface and the hydrophobic interaction of amphipathic alpha helical region in the C-terminus with the hydrophobic acyl chains in the cell membrane. CA-MA3 exhibited lower activity in antibacterial, antitumor, and vesicle-aggregating and disrupting activities than CA-MA and CA-MA2. This result suggested that the excessive beta-turn structure by Gly-Pro-Gly in CA-MA3 seems to interrupt the ion channel/pore formation on the lipid bilayer. It was concluded that the appropriate flexibility or beta-turn structure provided by the central hinge is responsible for the effective antibiotic activity of the antimicrobial peptides with the helix-hinge-helix structure. PMID- 10675501 TI - A new liposomal formulation for antisense oligodeoxynucleotides with small size, high incorporation efficiency and good stability. AB - Antisense oligodeoxynucleotides (asODN) are therapeutic agents that are designed to inhibit the expression of disease-related genes. However, their therapeutic use may be hindered due to their rapid clearance from blood and their inefficiency at crossing cell membranes. Cationic liposome complexes have been used to enhance the intracellular delivery of asODN in vitro; however, this type of carrier has unfavorable pharmacokinetics for most in vivo applications. Significant therapeutic activity of cationic liposomal asODN following systemic administration has not been demonstrated. In an effort to develop improved liposomal carriers for asODN for in vivo applications, we have evaluated the physical characteristics of two formulations which represent alternatives to cationic liposome-asODN complexes: asODN passively entrapped within neutral liposomes (PELA) and asODN formulated in a novel coated cationic liposomal formulation (CCL). Our results confirm that PELA can be extruded to small diameters that are suitable for intravenous administration. PELA are stable in human plasma; however, the incorporation efficiency is relatively low ( approximately 20%). The CCL formulation can also be extruded to small diameters (<200 nm), with significantly higher (80-100%) incorporation efficiency and are stable in 50% human plasma at 37 degrees C. A liposomal carrier for asODN with these characteristics may provide a significant therapeutic advantage over free asODN for some therapeutic applications. PMID- 10675502 TI - Interaction of cisplatin with planar model membranes - dose dependent change in electrical characteristics. AB - The drug cisplatin has broad antineoplastic activity against advanced testicular and ovarian cancers, epithelial malignancies, cancers of the head, neck, bladder, oesophagus and lungs. Peripheral neurotoxicity, ototoxicity and nephrotoxicity are its major side effects. The nonspecific action of this drug on the lipid bilayer architecture of membranes has been studied by following the effects produced on the electrical characteristics of model planar bilayer lipid membranes (BLM). The results confirm that the drug has a strong surface interaction with the zwitterionic polar head groups of the amphipathic phospholipids constituting the BLM. The permeability characteristics of cisplatin through the hydrophobic core are limited. Cisplatin does not fluidise the membrane sufficiently to cause its breakdown but creates small ion conducting defects on the membrane bilayer resulting in a marginal increase in ion conductivity. These results indicate that cisplatin exhibits a non-specific action on the lipid bilayer component of the membrane which might be partly responsible for its neurotoxic side effects. PMID- 10675503 TI - Beta(3)-adrenergic stimulation and insulin inhibition of non-selective cation channels in white adipocytes of the rat. AB - Single-channel currents were recorded from the plasma membrane of white adipocytes of 6-8-week-old male Sprague-Dawley rats. In outside-out patches (high K(+), no Ca(2+) in pipette), a voltage-dependent K-channel (delayed rectifier) with a single-channel conductance (gamma) of 16 pS (24 degrees C) in modified Ringer's was active at a density of 0.5/microm(2). It was blocked by TEA (IC(50)=1.5 mM). A Ca(2+)-activated non-selective cation channel (NSC-channel) appeared at a mean density of 1/microm(2) in inside-out patches ([Ca(2+)](i)=1.2 mM). gamma was 28 pS (24 degrees C). The NSC showed weak voltage dependence and was blocked by mefenamic acid and by internal ATP. In the cell-attached mode spontaneous activity could be blocked reversibly by 100 nM insulin. Noradrenaline (NA, 100 nM) induced a flickering activity of the NSC-channels. Isoproterenol (100 nM) caused activity of the NSC-channel as well. After 1 microM propranolol even 1 microM NA did not induce any activity. The alpha-antagonist phentolamine had no effect on isoproterenol- or on NA-induced currents. The beta(3)-agonists BRL 37344 and BRL 35135A induced activity of the NSC-channel at 100 nM as well. We conclude that white adipocytes express ion channels which are comparable to those in brown adipocytes and that beta-receptor activation opens NSC-channels thus allowing for Na(+) entry into white adipocytes. PMID- 10675504 TI - Demonstration of a fusion mechanism between a fluid bactericidal liposomal formulation and bacterial cells. AB - It was previously demonstrated that fluid liposomal-encapsulated tobramycin, named Fluidosomes, was successful in eradicating mucoid Pseudomonas aeruginosa in an animal model of chronic pulmonary infection, whereas free antibiotic did not reduce colony-forming unit (CFU) counts (C. Beaulac et al., Antimicrob. Agents Chemother. 40 (1996) 665-669; C. Beaulac et al., J. Antimicrob. Chemother. 41 (1998) 35-41). These liposomes were also shown to be bactericidal in in vitro tests against strong resistant P. aeruginosa 64 microg/ml). The time needed to reach the maximal fusion rate was about 5 h for the resistant strain comparatively to much shorter time for the sensitive strain. The specific characteristics of Fluidosomes could help overcome bacterial resistance related to permeability barrier and even enzymatic hydrolysis considering the importance of synergy in the whole process of antibiotic resistance. PMID- 10675505 TI - The LEA-like protein HSP 12 in Saccharomyces cerevisiae has a plasma membrane location and protects membranes against desiccation and ethanol-induced stress. AB - The LEA-like protein HSP 12 was identified as having a plasma membrane location in yeast. Gold particles, indicative of the presence of HSP 12, were observed on the external side of the plasma membrane when yeast grown to stationary phase were subjected to immunocytochemical analysis. Growth of yeast in the osmolyte mannitol resulted in an increased number of gold particles that were now observed to be present on both sides of the plasma membrane. No gold particles were observed using a mutant strain of the same yeast that did not express HSP 12. A model liposome system encapsulating the fluorescent dye calcein was used to investigate the protection by HSP 12 of membranes during desiccation. HSP 12 was found to act in an analogous manner to trehalose and protect liposomal membrane integrity against desiccation. The interaction between HSP 12 and the liposomal membrane was judged to be electrostatic as membrane protection was only observed with positively charged liposomes and not with either neutral or negatively charged liposomes. The ability of the wild-type and mutant yeast to grow in media containing ethanol was compared. It was found that yeast not expressing the HSP 12 protein were less able to grow in media containing ethanol. HSP 12 was shown to confer increased integrity on the liposomal membrane in the presence of ethanol. Ethanol, like mannitol, was found to induce HSP 12 protein synthesis. However, yeast grown in both ethanol and mannitol showed a decreased HSP 12 response compared with yeast grown in the presence of either osmolyte alone. PMID- 10675506 TI - Calcium enhances the transfection potency of plasmid DNA-cationic liposome complexes. AB - It is shown that calcium increases the in vitro transfection potency of plasmid DNA-cationic liposome complexes from 3- to 20-fold. The effect is Ca(2+) specific as other cations, such as Mg(2+) and Na(+), do not give rise to enhanced transfection and the effect can be inhibited by the presence of EGTA. It is shown that Ca(2+) increases cellular uptake of the DNA-lipid complexes, indicating that increased transfection potency arises from increased intracellular delivery of both cationic lipid and plasmid DNA in the presence of Ca(2+). In particular, it is shown that the levels of intact intracellular plasmid DNA are significantly enhanced when Ca(2+) is present. The generality of the Ca(2+) effect for enhancing complex-mediated transfection is demonstrated for a number of different cell lines and different cationic lipid formulations. It is concluded that addition of Ca(2+) represents a simple and useful protocol for enhancing in vitro transfection properties of plasmid DNA-cationic lipid complexes. PMID- 10675507 TI - Saturable ethanol binding in rat liver mitochondria. AB - The binding of ethanol to rat liver mitochondria is shown to be saturable at physiologically relevant ethanol concentrations. This effect is reversible and is not observed in extracted mitochondrial phospholipids. Brief exposure of the mitochondria to heat abolishes saturable ethanol binding. Previously, saturable ethanol binding was reported in rat liver microsomes. Taken together, the studies indicate that saturable ethanol binding motifs may be widespread in cellular membranes. The possibility is raised that incomplete expression of the hydrophobic effect in membrane assembly results in the expression of amphipathic packing defects which display an affinity for and a sensitivity to ethanol. The presence of saturable binding modalities is reconciled with the long-standing consensus on the biodistribution of ethanol - that ethanol's interactions with tissue are negligible - on the grounds that the affinities of ethanol and of water for membranes are similar; consequently, free ethanol concentrations are insensitive to the presence of tissue despite significant ethanol binding. A fraction of the binding sites possess submillimolar affinities for ethanol consistent with published functional studies, both in vitro and in vivo, that reported submillimolar efficacies for ethanol. PMID- 10675508 TI - Fusion of vesicles with the air-water interface: the influence of polar head group, salt concentration, and vesicle size. AB - Fusion of vesicles with the air-water interface and consequent monolayer formation has been studied as a function of temperature. Unilamellar vesicles of DMPC, DPPC, and DODAX (X=Cl(-), Br(-)) were injected into a subphase containing NaCl, and the surface pressure (tension) was recorded on a Langmuir Balance (Tensiometer) using the Wilhelmy plate (Ring) method. For the zwitterionic vesicles, plots of the initial surface pressure increase rate (surface tension decrease rate) as a function of temperature show a peak at the phase transition temperature (T(m)) of the vesicles, whereas for ionic ones they show a sharp rise. At high concentrations of NaCl, ionic DODA(Cl) vesicles seem to behave like zwitterionic ones, and the rate of fusion is higher at the T(m). The influence of size was studied comparing large DODA(Cl) vesicles with small sonicated ones, and no significant changes were found regarding the rate of fusion with the air-water interface. PMID- 10675509 TI - Cardiolipin, alpha-D-glucopyranosyl, and L-lysylcardiolipin from gram-positive bacteria: FAB MS, monofilm and X-ray powder diffraction studies. AB - Cardiolipin preparations from Streptococcus B, Listeria welshimeri, Staphylococcus aureus, and a glucosyl and lysyl derivative of cardiolipin were analysed for fatty acid composition and fatty acid combinations. Three different fatty acid patterns are described and up to 17 molecular species were identified in Streptococcus B lipids by high resolution FAB MS. The physicochemical properties of these lipids were characterised in the sodium salt form by monofilm experiments and X-ray powder diffraction. All lipids formed stable monofilms. The minimal space requirement of unsubstituted cardiolipin was dictated by the fatty acid pattern. Substitution with L-lysine led to a decrease of the molecular area, substitution with D-glucopyranosyl to an increase. On self assembly at 100% relative humidity, all preparations adopted lamellar structures. They showed a high degree of order, in spite of the heterogeneous fatty acid compositions and numerous fatty acid combinations. The repeat distances in lamellar fluid phase varied between 4.99 and 5. 52 nm, the bilayer thickness between 3.70 and 4.46 nm. Surprising were the low values of sorbed water per molecule of the glucosyl and lysyl derivatives which were 58 and 60%, compared with those of the respective cardiolipin. When Na(+) was replaced as counterion by Ba(2+), the bilayer structure was retained, but the lipids were in the lamellar gel phase and the fatty acids were tilted between 32 and 53 degrees away from the bilayer normal. Wide angle X-ray diffraction studies and electron density profiles are also reported. Particular properties of glucosyl cardiolipin are discussed. PMID- 10675510 TI - Deuterium NMR investigation of an amphotericin B derivative in mechanically aligned lipid bilayers. AB - The methyl-d(3) amide derivative of the polyene antibiotic amphotericin B was synthesized, assayed for biological activity, incorporated into mechanically aligned bilayers of dipalmitoylphosphatidylcholine (DPPC), and examined by deuterium and phosphorus NMR. The amide derivative has a lesser, but qualitatively similar, biological activity relative to amphotericin B. Incorporation of the amide derivative and ergosterol into aligned DPPC bilayers resulted in a single, stable bilayer phase, as shown by phosphorus NMR of the DPPC headgroups. Deuterium NMR spectra revealed one major (2)H quadrupolar splitting and one major (2)H-(1)H dipolar splitting in the liquid-crystalline phase, consistent with a high degree of alignment and a single, averaged physical state for amphotericin B methyl-d(3) amide in the bilayer. Variations of the quadrupolar and dipolar splittings as a function of macroscopic sample orientation and temperature indicated that the amide derivative undergoes fast rotation about a motional axis that is parallel to the bilayer normal. PMID- 10675511 TI - Efficient gene transfer by transferrin lipoplexes in the presence of serum. AB - Cationic lipids are being used increasingly as reagents for gene delivery both in vitro and in vivo. One of the limitations to the application of cationic lipid DNA complexes (lipoplexes) in vivo is the inhibition of gene delivery by serum. In this study, we have shown that transferrin (Tf)-lipoplexes, which had transferrin adsorbed at their surface via electrostatic interactions, are much more effective than plain lipoplexes in transfecting cells in the presence of relatively high concentrations (up to 60%) of fetal bovine serum (FBS). Serum even enhanced transfection by Tf-lipoplexes composed of 1,2-dioleoyl-3 (trimethylammonium) propane (DOTAP)/dioleoylphosphatidylethanolamine (DOPE)/pCMVLacZ at high lipid/DNA (+/-) charge ratios, and inhibited lipofection for those with low charge ratios when they were added to the cells immediately after the preparation of complexes. The effect of serum on lipofection was dose dependent. Preincubation of the complexes at 20 degrees C for 6 h led to serum resistance, even for the negatively charged transferrin-lipoplexes. A similar tendency was observed for DOTAP/cholesterol and DOTAP/DOPE/cholesterol liposomes. The percentage of cells transfected, measured by beta-galactosidase expression, also increased with the serum concentration. Cell viability was not affected significantly when the cells were incubated with the complexes for 4 h at 37 degrees C, followed by a 48-h incubation. Our findings extend the scope of previous studies where transferrin-lipoplexes were used to introduce DNA into cells, rendering these complexes and their future derivatives potential alternatives to viral vectors for gene delivery in vivo. PMID- 10675513 TI - Nucleotide chain length and the morphology of complexes with cationic amphiphiles: (31)P-NMR observations. AB - 31P-NMR and UV spectroscopies were used to study the interactions between cationic amphiphile-containing lipid bilayers and either a phosphorothioate oligonucleotide (OligoS) (n=21) or polyadenylic acid (PolyA) (n approximately 18,000). Multilamellar vesicles (MLVs) were composed of 1-palmitoyl-2-oleoyl-sn glycero-3-phosphocholine (POPC) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) in binary mixture with either of the cationic lipids, N-[1-(2, 3 dioleoyloxy)propyl]-N',N',N'-trimethylammonium chloride (DOTAP) or cetyltrimethylammonium bromide (CTAB). A UV-difference assay showed that OligoS binding ceased above a 1:1 anion/cation ratio, while PolyA binding continued until a 2:1 ratio was reached, indicating a 'flat' conformation for bound OligoS, but not necessarily for PolyA. Cross-polarization (31)P-NMR of the nucleotide chains bound to 100% DOTAP MLVs produced spectra virtually identical to those of dry powders of OligoS or PolyA, indicating effective immobilization of the surface-bound nucleotide chains. Hahn echo (31)P-NMR showed that MLVs composed of binary mixtures of POPC with DOTAP or CTAB retained a lamellar bilayer architecture upon adding nucleotide chains. At less than stoichiometric anion/cation ratios little or no signal attributable to free nucleotide chains was visible. A narrow signal at the chemical shift expected for phosphorothiodiesters or phosphodiesters became visible at greater levels of added OligoS or PolyA, respectively, indicating the presence of mobile nucleotide chains. Salt addition caused complete desorption of the nucleotide chains. When POPC was replaced with DOPE, binding of OligoS or PolyA produced non-bilayer lipid phases in the presence of DOTAP, but not in the presence of CTAB. PMID- 10675512 TI - A peptide analogue to a fusion domain within photoreceptor peripherin/rds promotes membrane adhesion and depolarization. AB - Photoreceptor peripherin/rds promotes membrane fusion, through a putative fusion domain located within the C-terminus (Boesze-Battaglia et al., Biochemistry 37 (1998) 9477-9487). A peptide analogue to this region, PP-5, competitively inhibits peripherin/rds mediated fusion in a cell free assay system. To characterize how this region is involved in the fusion process we investigated two of the individual steps in membrane fusion, membrane adhesion and membrane destabilization inferred from depolarization studies. Membrane depolarization was measured as the collapse of a valinomycin induced K(+) diffusion potential in model membranes, using a potential sensitive fluorescent probe, diS-C(2)-5. PP-5 induced membrane depolarization in a concentration dependent manner. PP-5 has been shown by Fourier transform infrared spectroscopy to be an amphiphilic alpha helix. Therefore, the requirement for an amphiphilic alpha-helix to promote depolarization was tested using two mutant peptides designed to disrupt either the amphiphilic nature of PP-5 (PP-5AB) or the alpha-helical structure (PP-5HB). PP-5AB inhibited PP-5 induced depolarization when added in an equimolar ratio to PP-5. Neither mutant peptide alone or in combination with PP-5 had any effect on calcium dependent vesicle aggregation. Using non-denaturing gel electrophoresis and size exclusion chromatography techniques PP-5 was shown to form a tetrameric complex. Equimolar mixtures of PP-5 and PP-5AB formed a heterotetramer which was unable to promote membrane depolarization. The hypothesis that PP-5 tetramers promote membrane depolarization is consistent with the calculated Hill coefficient of 3.725, determined from a Hill analysis of the depolarization data. PMID- 10675514 TI - Molecular cloning and expression of aquaporin 1 [correction of aquapolin 1] (AQP1) in dog kidney and erythroblasts. AB - Complementary DNA of the water channel aquaporin 1 (AQP1) was cloned from dog kidney and erythroblasts. The cDNA amplified from mRNA in dog kidney was 816 bp, the same as that in bovines, but longer by 6 bp than that in humans, mice and rats. The 235-bp fragment cDNA amplified from the mRNA in dog erythroblasts, which was differentiated from peripheral blood, was completely identical to the corresponding sequence of cDNA from the dog kidney. Thus, mature red blood cells from dog may have AQP1 in their cell membranes. The amino acid sequence in dog AQP1 was 91-94% identical to that in the other species mentioned above. Dog AQP1 has six predicted transmembrane domains, two NPA motifs, one mercury-sensitive site and four consensus phosphorylation sites, the same as the other species. However, dog and bovine AQP1 have only one N-glycosylation site, while two glycosylation sites were found in human and rodent AQP1. Xenopus oocytes injected with the mRNA of the dog AQP1 exhibited high water permeability in a hyposmotic medium. Thus, dog AQP1 performs water transport the same as in the other species. PMID- 10675515 TI - In vitro characterization of a novel polymeric-based pH-sensitive liposome system. AB - This study demonstrates rapid and pH-sensitive release of a highly water-soluble fluorescent aqueous content marker, pyranine, from egg phosphatidylcholine liposomes following incorporation of N-isopropylacrylamide (NIPA) copolymers in liposomal membranes. The pH-sensitivity of this system correlates with the precipitation of the copolymers at acidic pH. In vitro release can be significantly improved by increasing the percentage of anchor in the copolymer and thus favoring its binding to the liposomal bilayer. In the case of liposomes containing a poly(ethylene glycol)-phospholipid conjugate, the insertion of the pH-sensitive copolymer in the liposomal membrane appears to be sterically inhibited. Dye release from these formulations at acidic pH can still be achieved by varying the anchor molar ratio and/or molecular mass of the polymers or by including the latter during the liposome preparation procedure. Removal of unbound polymer results in decreased leakage only when the copolymer is inserted by incubation with preformed liposomes, but can be overcome by preparing liposomes in the presence of polymer. Aqueous content and lipid mixing assays suggest contents release can occur without membrane fusion. The results of this study indicate that the addition of pH-sensitive copolymers of NIPA represents promising strategy for improving liposomal drug delivery. PMID- 10675516 TI - Liquid crystalline/gel state phase separation in docosahexaenoic acid-containing bilayers and monolayers. AB - The phase behavior of lipid mixtures containing 1-stearoyl-2-docosahexaenoyl-sn glycero-3-phosphocholine (18:0, 22:6 PC) with 1,2-dipalmitoyl-sn-glycero-3 phosphocholine (DPPC) was studied with bilayers using differential scanning calorimetry (DSC), and with monolayers monitoring pressure/area isotherms and surface elasticity, and lipid domain formation followed by epifluorescence microscopy. From DSC studies it is concluded that DPPC/18:0, 22:6 PC phase separates into DPPC-rich and 18:0, 22:6 PC-rich phases. In monolayers, phase separation is indicated by changes in pressure-area isotherms implying phase separation where 18:0, 22:6 PC is 'squeezed out' of the remaining DPPC monolayer. Phase separation into lipid domains in the mixed PC monolayer is quantified by epifluorescence microscopy using the fluorescently labeled phospholipid membrane probe, 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl). These results further describe the ability of docosahexaenoic acid to participate in lipid phase separations in membranes. PMID- 10675517 TI - Properties of a reconstituted eukaryotic hexose/proton symporter solubilized by structurally related non-ionic detergents: specific requirement of phosphatidylcholine for permease stability. AB - Overexpression of the hexose/proton symporter HUP1 from Chlorella kessleri in S. cerevisiae permits a one-step purification via a biotinylation domain. Milligram amounts of the protein are obtained starting from 2 l of yeast culture. The HUP1 protein is used as a model eukaryotic membrane protein of the 'major facilitator superfamily' (MFS) to study specific lipid requirements for activity and stability. Testing two series of detergents revealed that n-nonyl-beta-D glucoside (NG) and n-octyl-beta-D-glucoside (OG) solubilize the HUP1 protein efficiently. Only the use of NG resulted in long-term stabilization of the HUP1 protein in the absence of external lipids. When affinity purified protein was extracted with organic solvents, a stoichiometric amount of phosphatidyl choline, phosphatidyl ethanolamine and ergosterol in the ratio of close to 2:1 was detected. These lipids were only observed, however, when the protein purification was carried out in the presence of NG; no lipids were copurified with the HUP1 protein in the presence of OG. Of the three lipids copurified, phosphatidyl choline showed a crucial role in ensuring maximal HUP1 permease activity and stability when added back to the OG-protein. The requirement of phosphatidylcholine documents a specific effect of lipids on vectorial transport mediated by a eukaryotic membrane protein of the MFS family. PMID- 10675518 TI - Selective destabilization of acidic phospholipid bilayers performed by the hepatitis B virus fusion peptide. AB - A peptide corresponding to the N-terminal region of the S protein of hepatitis B virus (Met-Glu-Asn-Ile-Thr-Ser-Gly-Phe-Leu-Gly-Pro-Leu-Leu-Val-Leu-Gln) has been previously demonstrated to perform aggregation and destabilization of acidic liposome bilayers and to adopt a highly stable beta-sheet conformation in the presence of phospholipids. The changes in the lipid moiety produced by this peptide have been followed by fluorescence depolarization and electron microscopy. The later was employed to determine the size and shape of the peptide vesicle complexes, showing the presence of highly aggregated and fused structures only when negatively charged liposomes were employed. 1,6-Diphenyl-1,3,5 hexatriene depolarization measurements showed that the interaction of the peptide with both negatively charged and zwitterionic liposomes was accompanied by a substantial reduction of the transition amplitude without affecting the temperature of the gel-to-liquid crystalline phase transition. These data are indicative of the peptide insertion inside the bilayer of both types of liposomes affecting the acyl chain order, though only the interaction with acidic phospholipids leads to aggregation and fusion. This preferential destabilization of the peptide towards negatively charged phospholipids can be ascribed to the electrostatic interactions between the peptide and the polar head groups, as monitored by 1-(4-(trimethylammoniumphenyl)-6-phenyl-1,3, 5-hexatriene fluorescence depolarization analysis. PMID- 10675519 TI - Redox- and pH-dependent association of plastocyanin with lipid bilayers: effect on protein conformation and thermal stability. AB - The effect of electrostatic interactions on the conformation and thermal stability of plastocyanin (Pc) was studied by infrared spectroscopy. Association of any of the two redox states of the protein with positively charged membranes at neutral pH does not significantly change the secondary structure of Pc. However, upon membrane binding, the denaturation temperature decreases, regardless of the protein redox state. The extent of destabilization depends on the proportion of positively charged lipid headgroups in the membrane, becoming greater as the surface density of basic phospholipids increases. In contrast, at pH 4.8 the membrane binding-dependent conformational change becomes redox sensitive. While the secondary structures and thermal stabilities of free and membrane-bound oxidized Pc are similar under acidic conditions, the conformation of the reduced form of the protein drastically rearranges upon membrane association. This rearrangement does not depend on electrostatic interactions to occur, since it is also observed in the presence of uncharged lipid bilayers. The conformational transition, only observed for reduced Pc, involves the exposure of hydrophobic regions that leads to intermolecular interactions at the membrane surface. Membrane-mediated partial unfolding of reduced Pc can be reversed by readjusting the pH to neutrality, in the absence of electrostatic interactions. This redox-dependent behavior might reflect specific structural requirements for the interaction of Pc with its redox partners. PMID- 10675520 TI - Effect of free fatty acids on the permeability of 1,2-dimyristoyl-sn-glycero-3 phosphocholine bilayer at the main phase transition. AB - We measured the influence of saturated and unsaturated free fatty acids on the permeability and partition of ions into 1, 2-dimyristoyl-sn-glycero-3 phosphocholine (DMPC) bilayers. The bilayer permeability was measured using the depletion of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1, 2-dihexadecanoyl-sn-glycero 3-phosphatidylethanolamine (N-NBD-PE) fluorescence as a result of its reduction by dithionite. We observed a distinct increase of dithionite permeability at the main gel-fluid phase transition of DMPC. When vesicles were formed from a mixture of DMPC and oleic acid, the membrane permeability at the phase transition was reduced drastically. Stearic acid and methyl ester of oleic acid have little effect. Similar results in the quenching of pyrene-PC in DMPC vesicles by iodide were obtained. Again, the increase of iodide partition into the lipid phase at the main phase transition of DMPC was abolished by the addition of unsaturated free fatty acids. Free fatty acids, in concentrations up to 5 mol%, do not abolish DMPC phase transition when measured by differential scanning calorimetry. It seems that unsaturated, but not saturated, free fatty acids reduce the lipid bilayer permeability to dithionite and iodide ions at the main phase transition of DMPC, without altering the thermodynamic properties of the bilayer. PMID- 10675521 TI - The plasma membrane NADH oxidase of soybean has vitamin K(1) hydroquinone oxidase activity. AB - Isolated plasma membrane vesicles and the plasma membrane NADH oxidase partially purified from soybean plasma membrane vesicles exhibited a cyanide-insensitive vitamin K(1) hydroquinone oxidase activity with isolated plasma membrane vesicles. Reduced vitamin K(1) (phylloquinol) was oxidized at a rate of about 10 nmol/min/mg protein as determined by reduced vitamin K(1) reduction or oxygen consumption. The K(m) for reduced K(1) was 350 microM. With the partially purified enzyme, reduced vitamin K(1) was oxidized at a rate of about 600 nmol/min/mg protein and the K(m) was 400 microM. When assayed in the presence of 1 mM KCN, activities of both plasma membrane vesicles and of the purified protein were stimulated (0.1 microM) or inhibited (0.1 mM) by the synthetic auxin growth factor 2, 4-dichlorophenoxyacetic acid. The findings suggest the potential participation of the plasma membrane NADH oxidase as a terminal oxidase of plasma membrane electron transport from cytosolic NAD(P)H via reduced vitamin K(1) to acceptors (molecular oxygen or protein disulfides) at the cell surface. PMID- 10675523 TI - Characterization of a cDNA encoding a rice mitochondrial voltage-dependent anion channel and its gene expression studied upon plant development and osmotic stress. AB - The voltage-dependent anion channel (VDAC) of mitochondria forms a large pore in the outer envelope membrane. Here, the full Oryza sativa OSVDAC1 cDNA was sequenced and is shown to belong to a small multigene family in the rice genome. This cDNA is 1093 bp long and codes for a protein of 274 amino acids. Expression studies of the osvdac1 gene show a regulation of its level in function of the plantlets maturation and organs. In contrast with several bacterial porins, osmotic stress does not have any effect on the plant osvdac1 gene expression. PMID- 10675522 TI - Human serum albumin enhances DNA transfection by lipoplexes and confers resistance to inhibition by serum. AB - Cationic liposome-DNA complexes ('lipoplexes') are used as gene delivery vehicles and may overcome some of the limitations of viral vectors for gene therapy applications. The interaction of highly positively charged lipoplexes with biological macromolecules in blood and tissues is one of the drawbacks of this system. We examined whether coating cationic liposomes with human serum albumin (HSA) could generate complexes that maintained transfection activity. The association of HSA with liposomes composed of 1, 2-dioleoyl-3-(trimethylammonium) propane and dioleoylphosphatidylethanolamine, and subsequent complexation with the plasmid pCMVluc greatly increased luciferase expression in epithelial and lymphocytic cell lines above that obtained with plain lipoplexes. The percentage of cells transfected also increased by an order of magnitude. The zeta potential of the ternary complexes was lower than that of the lipoplexes. Transfection activity by HSA-lipoplexes was not inhibited by up to 30% serum. The combined use of HSA and a pH-sensitive peptide resulted in significant gene expression in human primary macrophages. HSA-lipoplexes mediated significantly higher gene expression than plain lipoplexes or naked DNA in the lungs and spleen of mice. Our results indicate that negatively charged HSA-lipoplexes can facilitate efficient transfection of cultured cells, and that they may overcome some of the problems associated with the use of highly positively charged complexes for gene delivery in vivo. PMID- 10675524 TI - Expression and subcellular localization of a membrane protein related to Hsp30p in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae YDR033w gene product is homologous to Hsp30p and Yro2p, both of which are induced during heat shock. To investigate the subcellular localization of the YDR033w gene product, hemagglutinin (HA) epitope tagged protein was expressed, detected on immunoblots, and localized by immunofluorescence to cell membranes, primarily the plasma membrane. A punctuate immunofluorescence pattern was observed within cell buds. The nuclear envelope, but not the vacuole or mitochondrial membranes, were also immunostained. We refer to YDR033w as MRH1 to denote that it encodes a membrane protein related to Hsp30p. PMID- 10675525 TI - Is the manganese stabilizing 33 kDa protein of photosystem II attaining a 'natively unfolded' or 'molten globule' structure in solution? AB - This study compares the properties of the extrinsic 33 kDa subunit acting as 'manganese stabilizing protein' (MSP) of the water oxidizing complex with characteristic features of proteins that are known to attain a 'natively unfolded' or a 'molten globule' structure. The analysis leads to the conclusion that the MSP in solution is most likely a 'molten globule' with well defined compact regions of beta structure. The possible role of these structural peculiarities of MSP in solution for its function as important constituent of the WOC is discussed. PMID- 10675526 TI - Inhibition of chymase reduces vascular proliferation in dog grafted veins. AB - We investigated the effect of a chymase inhibitor Suc-Val-Pro-Phe(P)(OPh)(2) on the proliferation of the grafted vein in dog. By 28 days after the operation, the mean intimal area of the grafted vein in the placebo group was 3.24+/-0.32 mm(2). The intimal area of the grafted vein in the chymase inhibitor-treated group was reduced to 63.9%. In the placebo group, the activities of chymase and angiotensin converting enzyme in grafted vein were significantly increased 15- and 2-fold, respectively. In the chymase inhibitor-treated group, chymase activity in the grafted veins was decreased significantly. These findings suggest that inhibition of chymase appears useful for preventing vascular proliferation. PMID- 10675527 TI - First evidence and characterization of an uncoupling protein in fungi kingdom: CpUCP of Candida parapsilosis. AB - An uncoupling protein (UCP) was identified in mitochondria from Candida parapsilosis (CpUCP), a non-fermentative parasitic yeast. CpUCP was immunodetected using polyclonal antibodies raised against plant UCP. Activity of CpUCP, investigated in mitochondria depleted of free fatty acids, was stimulated by linoleic acid (LA) and inhibited by GTP. Activity of CpUCP enhanced state 4 respiration by decreasing DeltaPsi and lowered the ADP/O ratio. Thus, it was able to divert energy from oxidative phosphorylation. The voltage dependence of electron flux indicated that LA had a pure protonophoretic effect. The discovery of CpUCP proves that UCP-like proteins occur in the four eukaryotic kingdoms: animals, plants, fungi and protists. PMID- 10675528 TI - Prothymosin alpha fragmentation in apoptosis. AB - We observed fragmentation of an essential proliferation-related human nuclear protein prothymosin alpha in the course of apoptosis induced by various stimuli. Prothymosin alpha cleavage occurred at the DDVD(99) motif. In vitro, prothymosin alpha could be cleaved at D(99) by caspase-3 and -7. Caspase hydrolysis disrupted the nuclear localization signal of prothymosin alpha and abrogated the ability of the truncated protein to accumulate inside the nucleus. Prothymosin alpha fragmentation may therefore be proposed to disable intranuclear proliferation related function of prothymosin alpha in two ways: by cleaving off a short peptide containing important determinants, and by preventing active nuclear uptake of the truncated protein. PMID- 10675529 TI - Nitric oxide donors, nitrosothiols and mitochondrial respiration inhibitors induce caspase activation by different mechanisms. AB - We investigated to what extent different types of NO donors induce caspase activation by opening of the mitochondrial permeability transition pore (PTP) or inhibition of mitochondrial respiration. We found that nitrosothiols can directly open the PTP in isolated mitochondria and cause cytochrome c release, whereas NONOate donors can not. In macrophages nitrosothiols cause caspase activation that is blocked by cyclosporin A or calcium chelation, both of which prevent PTP opening, whereas caspase activation caused by NONOates is much less sensitive to these agents. Inhibitors of mitochondrial respiration did not promote PTP opening in isolated mitochondria, and although they cause caspase activation in macrophages, this activation was slower than that caused by NO donors, and was relatively insensitive to cyclosporin and calcium chelators suggesting that PTP opening was not involved. PMID- 10675530 TI - TAK1 mediates an activation signal from toll-like receptor(s) to nuclear factor kappaB in lipopolysaccharide-stimulated macrophages. AB - Stimulation of monocytes/macrophages with lipopolysaccharide (LPS) results in activation of nuclear factor-kappaB (NF-kappaB), which plays crucial roles in regulating expression of many genes involved in the subsequent inflammatory responses. Here, we investigated roles of transforming growth factor-beta activated kinase 1 (TGF-TAK1), a mitogen-activated protein kinase kinase kinase (MAPKKK), in the LPS-induced signaling cascade. A kinase-negative mutant of TAK1 inhibited the LPS-induced NF-kappaB activation both in a macrophage-like cell line, RAW 264.7, and in human embryonic kidney 293 cells expressing toll-like receptor 2 or 4. Furthermore, we demonstrated that endogenous TAK1 is phosphorylated upon simulation of RAW 264.7 cells with LPS. These results indicate that TAK1 functions as a critical mediator in the LPS-induced signaling pathway. PMID- 10675531 TI - High-molecular-weight kininogen is a binding protein for tissue prokallikrein. AB - Human tissue prokallikrein, a zymogen of the kallikrein-kinin system, circulates in plasma bound to neutrophils. Because plasma kininogens contribute to the assembly of kinin-generating components on blood cells, these proteins were assessed for their ability to complex the kallikrein precursor. Using ligand blot and direct binding assays, biotinylated prokallikrein was found to bind only to high-molecular-weight kininogen and not to the low-molecular-weight form. The interaction was specific, reversible, and saturable yielding an estimated dissociation constant K(D)=690 nM and a 1:1 stoichiometry. Specific kininogen binding of tissue prokallikrein also occurred at physiological plasma protein concentrations. These results provide the first evidence for a novel function of high-molecular-weight kininogen as a binding protein for tissue prokallikrein that could serve to localize the kallikrein precursor on the neutrophil surface. PMID- 10675532 TI - Distinct importin recognition properties of histones and chromatin assembly factors. AB - Synthesis of the protein components of nuclear chromatin occurs in the cytoplasm, necessitating specific import into the nucleus. Here, we report the binding affinities of the nuclear localisation sequence (NLS)-binding importin subunits for a range of histones and chromatin assembly factors. The results suggest that import of histones to the nucleus may be mediated predominantly by importin beta1, whereas the import of the other components probably relies on the conventional alpha/beta1 import pathway. Differences in recognition by importin beta1 were observed between histone H2A and the variant H2AZ, as well as between histone H3/4 with or without acetylation. The results imply that different histone variants may possess distinct nuclear import properties, with acetylation possibly playing an inhibitory role through NLS masking. PMID- 10675533 TI - Defect in modification at the anticodon wobble nucleotide of mitochondrial tRNA(Lys) with the MERRF encephalomyopathy pathogenic mutation. AB - A mitochondrial tRNA(Lys) gene mutation at nucleotide position 8344 is responsible for the myoclonus epilepsy associated with ragged-red fibers (MERRF) subgroup of mitochondrial encephalomyopathies. Here, we show that normally modified uridine at the anticodon wobble position remains unmodified in the purified mutant tRNA(Lys). We have reported a similar modification defect at the same position in two mutant mitochondrial tRNAs(Leu)(UUR) in another subgroup, mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), indicating this defect is common in the two kinds of tRNA molecules with the respective mutations of the two major mitochondrial encephalomyopathies. We therefore suggest the defect in the anticodon is responsible, through the translational process, for the pathogenesis of mitochondrial diseases. PMID- 10675535 TI - Rap1-suppressed tumorigenesis is concomitant with the interference in ras effector signaling. AB - Expression of Rap1 blocks epithelial growth factor-induced extracellular signal regulated kinases (ERKs) activation. However, recent studies demonstrated that Rap1 mediates ERKs activation induced by nerve growth factor. The anti-oncogenic effect of Rap1 has been reported but its mechanism remains unclear. To evaluate the correlation between the anti-transforming effect and the activation of ERKs, we transfected rap1 cDNA into Hep3B cells and selected stable transfectants. The Rap1 transfectants completely lost their intrinsic tumorigenicity in Balb/c nude mice. Both insulin and 12-O-tetradecanoyl phorbol-13-acetate (TPA)-stimulated ERK activations were also blocked. Our findings suggest that Rap1-suppressed tumorigenicity is concomitant with ERKs inhibition. PMID- 10675534 TI - Identification and characterization of functional subunits of Clostridium botulinum type A progenitor toxin involved in binding to intestinal microvilli and erythrocytes. AB - Clostridium botulinum type A hemagglutinin-positive progenitor toxin consists of three distinct components: neurotoxin (NTX), hemagglutinin (HA), and non-toxic non-HA (NTNH). The HA consists of four subcomponents designated HA1, 2, 3a and 3b. By employing purified toxin and GST-fusion proteins of each HA subcomponent, we found that the HA-positive progenitor toxin, GST-HA1 and GST-HA3b bind to human erythrocytes and microvilli of guinea pig upper small intestinal sections. The HA-positive progenitor toxin and GST-HA1 bind via galactose moieties, GST HA3b binds via sialic acid moieties. GST-2 and GST-3a showed no detectable binding. PMID- 10675536 TI - Drosophila MTN: a metazoan copper-thionein related to fungal forms. AB - Two Drosophila metallothioneins (MT) have been reported: MTN, a 40 residue peptide including 10 Cys, and MTO, a 43 residue peptide including 12 Cys. However, neither functional nor evolutionary analyses for either of the Drosophila MT are available. Here, heterologous expression of Mtn in Escherichia coli is reported. The metal binding abilities of the Cu- and Zn-MTN complexes conformed in vivo, as well as the features of the Cd- and Cu-aggregates produced by metal replacement in vitro, have been determined by atomic emission spectrometry, circular dichroism and electrospray ionization mass spectrometry. Primary structure relationships with other MT have been examined. The results indicate a close resemblance of MTN to fungal copper-thioneins. PMID- 10675537 TI - Novel substrate specificity of a membrane-bound beta-glycosidase from the hyperthermophilic archaeon Pyrococcus horikoshii. AB - A beta-glycosidase gene homolog of Pyrococcus horikoshii (BGPh) was successfully expressed in Escherichia coli. The enzyme was localized in a membrane fraction and solubilized with 2.5% Triton X-100 at 85 degrees C for 15 min. The optimum pH was 6.0 and the optimum temperature was over 100 degrees C, respectively. BGPh stability was dependent on the presence of Triton X-100, the enzyme's half-life at 90 degrees C (pH 6.0) was 15 h. BGPh has a novel substrate specificity with k(cat)/K(m) values high enough for hydrolysis of beta-D-Glcp derivatives with long alkyl chain at the reducing end and low enough for the hydrolysis of beta linked glucose dimer more hydrophilic than aryl- or alkyl-beta-D-Glcp. PMID- 10675538 TI - Tumor necrosis factor-induced lethal hepatitis: pharmacological intervention with verapamil, tannic acid, picotamide and K76COOH. AB - Tumor necrosis factor (TNF) induces hepatitis when injected in human beings or in rodents. The molecular mechanism by which TNF induces hepatic distress remains largely unknown, although induction of apoptosis of hepatocytes appears to be an essential step. In order to increase the therapeutic value of TNF, we have studied the protective activity of several molecules and found that four chemically totally different substances confer significant protection in the model of TNF-induced lethal hepatitis in mice sensitized with D-(+)-galactosamine (GalN), but not in mice sensitized with actinomycin-D (ActD) or against anti-Fas induced lethal hepatitis. Verapamil, a calcium-channel blocker, tannic acid, picotamide, a thromboxane A(2) receptor antagonist, and K76COOH, an inhibitor, amongst others, of complement, protected significantly against induction of lethality, release of the liver-specific enzyme alanine aminotransferase (ALT) and induction of apoptosis in the liver after TNF/GalN, except for K76COOH, which paradoxically increased ALT values after challenge, and which also protected against TNF/GalN in complement-deficient mice. The data suggest that activation of platelets and neutrophils, as well as induction of inflammation occur in the TNF/GalN model, but not in the TNF/ActD or anti-Fas models, in which direct induction of apoptosis of hepatocytes may be more relevant. The protective activity of the drugs may lead to an increase in therapeutic value of TNF. PMID- 10675539 TI - Biotic and abiotic stress can induce cystatin expression in chestnut. AB - A cysteine proteinase inhibitor (cystatin) from chestnut (Castanea sativa) seeds, designated CsC, has been previously characterized. Its antifungal, acaricide and inhibitory activities have allowed to involve CsC in defence mechanisms. The CsC transcription levels decreased during seed maturation and increased throughout germination, an opposite behavior to that shown by most phytocystatins. No inhibition of endogenous proteinase activity by purified CsC was found during the seed maturation or germination processes. CsC message accumulation was induced in chestnut leaves after fungal infection, as well as by wounding and jasmonic acid treatment. Induction in roots was also observed by the last two treatments. Furthermore, CsC transcript levels strongly raised, both in roots and leaves, when chestnut plantlets were subjected to cold- and saline-shocks, and also in roots by heat stress. All together, these data suggest that chestnut cystatin is not only involved in defence responses to pests and pathogen invasion, but also in those related to abiotic stress. PMID- 10675540 TI - Possible role of immune surveillance at the initial phase of metastasis produced by B16BL6 melanoma cells. AB - The relationship among the real-time trafficking of lung metastatic B16BL6 cells, metastatic potential, and the injected number of the cells was examined, since the smaller the number of tumor cells injected, the more clearly the immune defense may be observed. When 1x10(6) or 1x10(5) B16BL6 cells were injected into mice via the tail vein, both numbers of cells accumulated in the lung at a similar rate: there was an approximately 10-fold difference in the number of accumulated cells between the two doses. Elimination from the lung was not dependent on the cell number but on the proportion of accumulated cells. However, the injection of 1x10(4) cells resulted in lung accumulation less than one-tenth of that obtained with 1x10(5) cell injection. Metastasis was observed when 1x10(5) or 1x10(6) B16BL6 cells were injected, but not after injection of 1x10(4) cells. To clarify the roles of the immune defense system at the initial phase of metastasis, we challenged macrophage-depleted mice with 1x10(4) tumor cells. Treatment of mice with 2-chloroadenosine prior to the tumor cell challenge cancelled the suppression of not only metastasis but also the lung accumulation. Furthermore, the administration of 2-chloroadenosine following the tumor cell challenge had little effect on the metastatic potential. These results suggest that the immune surveillance whose action was obvious at the low dose of challenged tumor cells functions strongly at the initial phase but not at the advanced stages of the metastatic process, and that macrophages play an important role in the suppression of metastasis. PMID- 10675541 TI - LOX-1 mediates lysophosphatidylcholine-induced oxidized LDL uptake in smooth muscle cells. AB - A novel receptor for oxidized low-density lipoprotein (OxLDL), lectin-like OxLDL receptor (LOX-1), was cloned from endothelial cells. Since OxLDL is taken up by vascular smooth muscle cells (VSMC) in atheroma, we analyzed the inducible expression of LOX-1 in VSMC in the present study. Incubation of cultured bovine VSMC with lysophosphatidylcholine (LPC), an atherogenic component of OxLDL, increased the level of mRNA for LOX-1 in a dose- and time-dependent manner. Since LPC did not significantly change the half-life of LOX-1 mRNA, the induction seemed to occur at the transcriptional level. The induction accompanied an increase in the protein level of LOX-1 and activity of OxLDL uptake. Blocking antibody against LOX-1 significantly suppressed the enhanced uptake of OxLDL. Thus, LOX-1 is a major receptor for OxLDL in VSMC as in endothelial cells. The enhanced expression of LOX-1 by LPC suggests that OxLDL and LPC would progressively change the function of VSMC and accelerate atherogenesis in vivo. PMID- 10675542 TI - Kinetic and spectroscopic characterization of native and metal-substituted beta lactamase from Aeromonas hydrophila AE036. AB - Two metal ion binding sites are conserved in metallo-beta-lactamase from Aeromonas hydrophila. The ligands of a first zinc ion bound with picomolar dissociation constant were identified by EXAFS spectroscopy as one Cys, two His and one additional N/O donor. Sulfur-to-metal charge transfer bands are observed for all mono- and di-metal species substituted with Cu(II) or Co(II) due to ligation of the single conserved cysteine residue. Binding of a second metal ion results in non-competitive inhibition which might be explained by an alternative kinetic mechanism. A possible partition of metal ions between the two binding sites is discussed. PMID- 10675543 TI - Structural domains of the insulin receptor and IGF receptor required for dimerisation and ligand binding. AB - We investigated structural requirements for dimerisation and ligand binding of insulin/IGF receptors. Soluble receptor fragments consisting of N-terminal domains (L1/CYS/L2, L1/CYS/L2/F0) or fibronectin domains (F0/F1/F2, F1/F2) were expressed in CHO cells. Fragments containing F0 or F1 domains were secreted as disulphide-linked dimers, and those consisting of L1/CYS/L2 domains as monomers. None of these proteins bound ligand. However, when a peptide of 16 amino acids from the alpha-subunit C-terminus was fused to the C-terminus of L1/CYS/L2, the monomeric insulin and IGF receptor constructs bound their respective ligands with affinity only 10-fold lower than native receptors. PMID- 10675544 TI - The molecular basis for genetic polymorphism of human deoxyribonuclease II (DNase II): a single nucleotide substitution in the promoter region of human DNase II changes the promoter activity. AB - Deoxyribonuclease II (DNase II) levels in human vary depending on whether the individual has the DNASE2*H (high) allele or the DNASE2*L (low) allele. We examined the promoter activity of the 5'-flanking region of each of these alleles by transient transfection luciferase assay. DNASE2*H had 5-fold higher promoter activity than DNASE2*L in human hepatoma HepG2 cell. Comparison of the nucleotide sequences of the proximal promoter regions revealed a G to A transition at position -75; G and A residues were assigned to DNASE2*H and *L, respectively. Since no differences were found between the open reading frame sequences of these alleles, it is likely that the A-75G transition causes the allelic difference in the promoter activity of the gene, underlying the genetic polymorphism. PMID- 10675545 TI - Isolation of acein-2, a novel angiotensin-I-converting enzyme inhibitory peptide derived from a tryptic hydrolysate of human plasma. AB - We previously described a novel angiotensin-I-converting enzyme (ACE) inhibitory peptide, designated Acein-1, that was isolated from a tryptic hydrolysate of human plasma. We now report a second such inhibitory peptide, Acein-2 obtained from the same hydrolysate. The peptide was purified by gel filtration and cation exchange chromatography followed by reversed-phase gradient and isocratic high performance liquid chromatography. Acein-2 was found to be a tripeptide, Leu-Ile Tyr, which is thought to correspond to f(518-520) of human alpha2-macroglobulin. The synthetic tripeptide showed a potent dose-dependent inhibition of ACE, with an IC(50) value of 0.82 micromol/l. Lineweaver-Burk plots suggested that Acein-2 as well as the previously described Acein-1 are non-competitive inhibitors. PMID- 10675546 TI - Post-translational phosphorylation affects the IgE binding capacity of caseins. AB - IgE response specific to those molecular regions of casein that contain a major phosphorylation site was analyzed using native and modified caseins and derived peptides. This study included (i) the naturally occurring common variants A1 and A from beta- and alphas2-caseins, respectively, which were purified in the native form and then dephosphorylated, (ii) a purified rare variant D of alphas2-casein which lacks one major phosphorylation site, and (iii) the native and dephosphorylated tryptic fragment f(1-25) from beta-casein. Direct and indirect ELISA using sera from patients allergic to milk showed that the IgE response to caseins is affected by modifying or eliminating the major phosphorylation site. PMID- 10675547 TI - Involvement of CDSP 32, a drought-induced thioredoxin, in the response to oxidative stress in potato plants. AB - In animal cells, yeast and bacteria, thioredoxins are known to participate in the response to oxidative stress. We recently identified a novel type of plant thioredoxin named CDSP 32 for chloroplastic drought-induced stress protein of 32 kDa. In the present work, we measured comparable increases in the glutathione oxidation ratio and in the level of chlorophyll thermoluminescence, a specific marker for thylakoid lipid peroxidation in Solanum tuberosum plants subjected to drought or oxidative treatments (photooxidative stress, gamma irradiation and methyl viologen spraying). Further, substantial accumulations of CDSP 32 mRNA and protein were revealed upon oxidative treatments. These data show for the first time in plants the induction of a thioredoxin by oxidative stress. We conclude that CDSP 32 may preserve chloroplastic structures against oxidative injury upon drought. PMID- 10675548 TI - SNS/PN3 and SNS2/NaN sodium channel-like immunoreactivity in human adult and neonate injured sensory nerves. AB - Two tetrodotoxin-resistant voltage-gated sodium channels, SNS/PN3 and SNS2/NaN, have been described recently in small-diameter sensory neurones of the rat, and play a key role in neuropathic pain. Using region-specific antibodies raised against different peptide sequences of their alpha subunits, we show by Western blot evidence for the presence of these channels in human nerves and sensory ganglia. The expected fully mature 260 kDa component of SNS/PN3 was noted in all injured nerve tissues obtained from adults; however, for SNS2/NaN, smaller bands were found, most likely arising from protein degradation. There was increased intensity of the SNS/PN3 260 kDa band in nerves proximal to the site of injury, whereas it was decreased distally, suggesting accumulation at sites of injury; all adult patients had a positive Tinel's sign at the site of nerve injury, indicating mechanical hypersensitivity. Injured nerves from human neonates showed similar results for both channels, but neonate neuromas lacked the SNS2/NaN 180 kDa molecular form, which was strongly present in adult neuromas. The distribution of SNS/PN3 and SNS2/NaN sodium channels in injured human nerves indicates that they represent targets for novel analgesics, and could account for some differences in the development of neuropathic pain in infants. PMID- 10675549 TI - Roles of p38 MAPK, PKC and PI3-K in the signaling pathways of NADPH oxidase activation and phagocytosis in bovine polymorphonuclear leukocytes. AB - Stimulation of bovine polymorphonuclear leukocytes (PMN) with serum-opsonized zymosan (sOZ) induced the activation of p38 mitogen-activated protein kinase (MAPK), protein kinase C (PKC) and phosphatidylinositol 3-kinase (PI3-K) and sOZ induced O(2)(-) production was significantly attenuated by their inhibitors (SB203580 for p38 MAPK, GF109203X for PKC and wortmannin for PI3-K). They caused significant attenuation of sOZ-induced phosphorylation of p47phox as well. Flow cytometric analysis, however, revealed that SB203580 and wortmannin attenuated phagocytosis, but GF109203X facilitated it. The results suggest that p38 MAPK and PI3-K participated in both signaling pathways of NADPH oxidase activation (O(2)( ) production) and phagocytosis, and PKC participated in the signaling pathway of NADPH oxidase activation alone. PMID- 10675550 TI - Peroxisome proliferator-activated receptors are expressed in mouse bone marrow derived mast cells. AB - We examined the expression of peroxisome proliferator-activated receptors (PPARs) and the role of PPARs in cytokine production in mouse bone marrow-derived mast cells (mBMMCs). mBMMCs expressed PPARbeta strongly and gamma slightly, but not alpha. Activation of mBMMCs with antigen or calcium ionophore resulted in the increased expression of PPARgamma mRNA specifically. 15-Deoxy-Delta(12, 14) prostaglandin J(2) (15d-PGJ(2)) and troglitazone, all PPARgamma ligands, attenuated the antigen-induced cytokine production by mBMMCs. Carbaprostacyclin, a PPARbeta ligand, also inhibited cytokine production, whereas PPARalpha ligands did not. These results suggest that PPARbeta and gamma might be included in the negative regulation of mast cell activation. PMID- 10675551 TI - Expression of fatty acid-CoA ligase 4 during development and in brain. AB - Fatty acid utilization is initiated by fatty acid-CoA ligase, which converts free fatty acids into fatty acyl-CoA esters. We have cloned previously the human long chain fatty acid-CoA ligase 4 (FACL4), which is a central enzyme in controlling the free arachidonic acid level in cells and thereby regulating eicosanoid production. We report here the expression of this gene in tissues, particularly in different parts of the brain. We found that FACL4 encoded a 75 kDa enzyme and that there was a modified translation product expressed in the brain. FACL4 was expressed in early stages of development with a significant amount of FACL4 mRNA detected in an E7 mouse embryo. In addition, FACL4 was highly expressed in both adult and newborn mouse brain especially in the granule cells of the dentate gyrus and the pyramidal cell layer of CA1 in hippocampus, and the granular cell layer and Purkinje cells of the cerebellum. PMID- 10675552 TI - Identification of a stimulus-dependent DNase I hypersensitive site between the Ialpha and Calpha exons during immunoglobulin heavy chain class switch recombination. AB - The complete humoral response to foreign antigen depends upon two distinct recombination events within the heavy chain locus of immunoglobulin. The first recombination event takes place in what will become the antigen combining site of the antibody molecule, encoded by V, D and J segments. The second recombination event involves the looping-out of large spans of DNA which separate the various clusters of heavy chain exons which define the different immunoglobulin isotypes, or classes. While a great deal has been learned about the nature of the VDJ recombinase, very little is known about the nature of the class-switch recombinase. Using a cell system where class-switch recombination occurs primarily to the IgA locus, we have looked for stimulus-dependent changes in the chromatin structure of the IgA locus which might result from interactions between components of the recombinase and cis-elements within the region. We present evidence that strongly suggests that the class-switch recombinase interacts between the Ialpha and Calpha exons of IgA, just upstream of the highly reiterated DR1 and DR2 elements. However, although multiple potential SMAD-4 sites are located precisely within the DNase I hypersensitive site and 160 bp upstream of that site, we failed to detect any evidence of DNA/protein interactions near the hypersensitive site. Moreover, recombinant SMAD-3/4 proteins fail to interact with these sites with appreciable affinity in vitro. These data suggest that some other structural alteration at this site (e.g. RNA/DNA hybrid) may mediate the nuclease sensitivity. PMID- 10675553 TI - Characterization of Arabidopsis AtAMT2, a novel ammonium transporter in plants. AB - We have cloned and characterized the first member of a novel family of ammonium transporters in plants: AtAMT2 from Arabidopsis thaliana. AtAMT2 is more closely related to bacterial ammonium transporters than to plant transporters of the AMT1 family. The protein was expressed and functionally characterized in yeast. AtAMT2 transported ammonium in an energy-dependent manner. In contrast to transporters of the AMT1 family, however, AtAMT2 did not transport the ammonium analogue, methylammonium. AtAMT2 was expressed more highly in shoots than roots and was subject to nitrogen regulation. PMID- 10675554 TI - Regulated but not constitutive human respiratory syncytial virus (HRSV) P protein phosphorylation is essential for oligomerization. AB - Purified human respiratory syncytial virus (HRSV) P phosphoprotein from transfected HEp-2 cells is able to oligomerize forming tetramers. The bulk of constitutive P protein phosphorylation (99. 8%) (serine residues 116, 117, 119, 232 and 237) can be removed without affecting protein oligomerization. However, dephosphorylated P protein, produced in bacteria, is unable to oligomerize. This difference can be explained by a transient P protein phosphorylation, detected in HEp-2 cells, that could be essential for P protein oligomerization. PMID- 10675555 TI - X-ray crystal structure of the YM210W mutant reaction centre from Rhodobacter sphaeroides. AB - The X-ray crystal structure of a reaction centre from Rhodobacter sphaeroides with a mutation of tyrosine M210 to tryptophan (YM210W) has been determined to a resolution of 2.5 A. Structural conservation is very good throughout the body of the protein, with the tryptophan side chain adopting a position in the mutant complex closely resembling that of the tyrosine in the wild-type complex. The spectroscopic properties of the YM210W reaction centre are discussed with reference to the structural data, with particular focus on evidence that the introduction of the bulkier tryptophan in place of the native tyrosine may cause a small tilt of the macrocycle of the B(L) monomeric bacteriochlorophyll. PMID- 10675556 TI - Preferential induction of apoptosis of leukaemic cells by farnesol. AB - Farnesol preferentially inhibits proliferation and induces apoptosis of tumour derived but not non-transformed cell lines. We investigated whether farnesol induces apoptosis of blasts from patients with acute myeloid leukaemia (AML) and leukaemic cell lines, as compared with normal, human primary haemopoietic cells. We show that 30 microM farnesol causes apoptosis of leukaemic cell lines of T- and B-lymphocyte, myeloid or erythroid lineages and primary blasts obtained from patients with AML. However, the same concentration did not kill primary monocytes, or quiescent or proliferating T-lymphocytes. We conclude that farnesol selectively kills AML blasts and leukaemic cell lines in preference to primary haemopoietic cells. PMID- 10675557 TI - Isolation of the epithiospecifier protein from oil-rape (Brassica napus ssp. oleifera) seed and its characterization. AB - The epithiospecifier protein (ESP) is a myrosinase (MYR) cofactor, which is necessary to drive the MYR-catalyzed hydrolysis of some specific glucosinolates towards the production of cyanoepithioalkanes instead of isothiocyanates and nitriles. ESP was isolated from Brassica napus seeds by anionic exchange and gel filtration chromatography. ESP showed a molecular weight of about 39 kDa and pI 5.3. The amino acid sequence of several tryptic peptides of ESP (accounting for about 50% of the total sequence) made it possible to establish the high similarity (81% identity) with a hypothetical 37 kDa protein (TrEMBL data base accession number Q39104) and several jasmonate-inducible proteins from Arabidopsis thaliana. This observation suggests that ESP is likely to be involved in jasmonate-mediated defence and disease resistance mechanisms. PMID- 10675558 TI - Identification of syntenin and other TNF-inducible genes in human umbilical arterial endothelial cells by suppression subtractive hybridization. AB - Endothelial cells play an important regulatory role in inflammatory responses by upregulating various proinflammatory gene products including cytokines and adhesion molecules. A highly potent mediator of this process is tumor necrosis factor-alpha (TNF). In the present study, the suppression subtractive hybridization (SSH) method was employed to identify rarely transcribed TNF inducible genes in human umbilical arterial endothelial cells. Following mRNA isolation of non-stimulated and TNF-stimulated cells, cDNAs of both populations were prepared and subtracted by suppression PCR. Sequencing of the enriched cDNAs identified 12 genes differentially expressed including vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, interleukin-8 and IkappaBalpha, an inhibitor of the transcription factor nuclear factor-kappaB. Interestingly, also syntenin, a PDZ motif-containing protein which binds to the cytoplasmic domain of syndecans, was identified by SSH. Time course studies using RT-PCR analysis confirmed that all genes were differentially expressed and rapidly induced by TNF. Our data reveal that SSH is a powerful technique of high sensitivity for the detection of differential gene expression in primary arterial endothelial cells. PMID- 10675559 TI - Inhibition of plant telomerase by telomere-binding proteins from nuclei of telomerase-negative tissues. AB - The activity of telomerase in plant cells is precisely regulated in response to changes in cell division rate. To explore this regulatory mechanism, the effect on telomerase activity of protein extracts from nuclei of telomerase-negative tissues was examined. An inhibition of telomerase activity was found which was species-non-specific. This inhibition was due to proteins which form salt-stable, sequence-specific complexes with the G-rich telomeric strand and reduce its accessibility, as shown by gel retardation and by terminal transferase (TdT) extension of G-rich telomeric and non-telomeric (substrate) primers. A 40 kDa polypeptide was detected by SDS-PAGE after cross-linking the complex formed by extracts from tobacco leaf nuclei. Such proteins may be involved in regulation of telomerase activity in plants. PMID- 10675560 TI - The Saccharomyces cerevisiae DNA damage checkpoint is required for efficient repair of double strand breaks by non-homologous end joining. AB - In this work we report that the Saccharomyces cerevisiae RAD9, RAD24, RAD17, MEC1, MEC3 and RAD53 checkpoint genes are required for efficient non-homologous end joining (NHEJ). RAD9 and RAD24 function additionally in this process. Defective NHEJ in rad9Delta-rad24Delta, but not yku80Delta cells, is only partially rescued by imposing G1 or G2/M delays. Thus, checkpoint functions other than transient cell cycle delays may be required for normal levels of NHEJ. Epistasis analysis also indicated that YKU80 and RAD9/RAD24 function in the same pathway for repair of lesions caused by MMS and gamma-irradiation. Unlike NHEJ, the checkpoint pathway is not required for efficient site-specific integration of plasmid DNA into the yeast genome, which is RAD52-dependent, but RAD51 independent. PMID- 10675561 TI - Analysis of the interaction between single-chain variable fragments and their antigen in a reducing intracellular environment using the two-hybrid system. AB - The coding sequences of three single-chain variable (scFv) fragments (A4, G4 and H3), which bind to dihydroflavonol-4-reductase (DFR) of Petunia hybrida, and the DFR-encoding sequence were cloned in two-hybrid vectors. The vectors were transformed in the yeast strain HF7c (his3-200, trp1-901, leu2-3) and the scFv DFR interaction was analyzed by measuring yeast growth on medium without histidine. ScFv-G4 and, to a lesser extent, scFv-A4 could interact with DFR in the yeast nucleus. On the contrary, scFv-H3 showed no interaction with its antigen in yeast. The results of a previous expression analysis of the same scFv fragments in the plant cytosol correlate with those of the two-hybrid test. This suggests that it is possible to evaluate the antigen-scFv interaction in a reducing subcellular environment with the two-hybrid test. Therefore, the yeast two-hybrid system can be useful to identify candidate scFv fragments for intracellular antibody applications. PMID- 10675562 TI - A novel target of lithium therapy. AB - Phosphatases converting 3'-phosphoadenosine 5'-phosphate (PAP) into adenosine 5' phosphate are of fundamental importance in living cells as the accumulation of PAP is toxic to several cellular systems. These enzymes are lithium-sensitive and we have characterized a human PAP phosphatase as a potential target of lithium therapy. A cDNA encoding a human enzyme was identified by data base screening, expressed in Escherichia coli and the 33 kDa protein purified to homogeneity. The enzyme exhibits high affinity for PAP (K(m)<1 microM) and is sensitive to subtherapeutic concentrations of lithium (IC(50)=0.3 mM). The human enzyme also hydrolyzes inositol-1, 4-bisphosphate with high affinity (K(m)=0.4 microM), therefore it can be considered as a dual specificity enzyme with high affinity (microM range) for both PAP and inositol-1,4-bisphosphate. Hydrolysis of inositol 1,4-bisphosphate was also inhibited by lithium (IC(50)=0.6 mM). Thus, we present experimental evidence for a novel target of lithium therapy, which could explain some of the side effects of this therapy. PMID- 10675563 TI - Phospholipase D2: functional interaction with caveolin in low-density membrane microdomains. AB - Low-density detergent-insoluble membrane domains contain caveolin-1 and are enriched in a phospholipase D activity that is not PLD1. Here we show that caveolin-rich fractions, prepared from HaCaT human keratinocytes by either detergent-based or detergent-free methods, contain PLD2. Caveolar membrane PLD activity is stimulated 2-fold by low concentrations (10-30 microM) of the caveolin-1 and caveolin-2 scaffolding domain peptides, whereas it is inhibited at higher concentrations of the peptides. Immunoisolated HA-tagged PLD1 and PLD2 are not stimulated by the peptides, although both enzymes retain sensitivity to their inhibitory effect. Down-regulation of caveolin-1 expression by treatment of the cells with acetyl-leucyl-leucyl-norleucinal decreased caveolar PLD activity by 50%. Similarly, expression of an active form of the sterol regulatory element binding protein (SREBP(1-490)) down-regulated caveolin-1 expression by 50% and decreased caveolar PLD activity by 60%. These data identify the PLD activity in caveolin-rich membranes as PLD2 and provide in vivo evidence suggesting that caveolin-1 regulates PLD2 activity. PMID- 10675564 TI - Characterization of a novel endopolygalacturonase from Aspergillus niger with unique kinetic properties. AB - We isolated and characterized a new type of endopolygalacturonase (PG)-encoding gene, pgaD, from Aspergillus niger. The primary structure of PGD differs from that of other A. niger PGs by a 136 amino acid residues long N-terminal extension. Biochemical analysis demonstrated extreme processive behavior of the enzyme on oligomers longer than five galacturonate units. Furthermore, PGD is the only A. niger PG capable of hydrolyzing di-galacturonate. It is tentatively concluded that the enzyme is composed of four subsites. The physiological role of PGD is discussed. PMID- 10675565 TI - Molecular cloning of Drosophila gamma-glutamylcysteine synthetase by functional complementation of a yeast mutant. AB - gamma-Glutamylcysteine synthetase (GCS) catalyses a critical, rate-limiting step in glutathione synthesis. In this study we describe the isolation and characterisation of a GCS cDNA (pDmGCS4.3. 3) from Drosophila melanogaster by functional complementation of a Saccharomyces cerevisiae gsh1 mutant. Expression of pDmGCS4.3.3 in the yeast mutant partially restored glutathione levels and conferred resistance to methylglyoxal. The pDmGCS4.3.3 cDNA was found to be approx. 4.6 kb in length, containing a 2 kb fragment encoding an open reading frame with a high degree of deduced amino acid sequence identity with previously reported GCS sequences. In situ hybridisation revealed that the Drosophila GCS gene maps to 7D6-9 on the X chromosome. PMID- 10675566 TI - A GATA binding site is involved in the regulation of 15-lipoxygenase-1 expression in human colorectal carcinoma cell line, caco-2. AB - The data presented implicate a GATA binding site in the transcriptional regulation of 15-lipoxygenase-1 (15-LO-1) gene expression in human colorectal carcinoma Caco-2 cells. High expression of GATA-6 mRNA and protein was observed, while GATA-4 mRNA was expressed at a very low level in Caco-2 cells. The expression of GATA-6 was down-regulated, while 15-LO-1 expression was dramatically up-regulated after treatment with sodium butyrate (NaBT). A study using an electrophoretic mobility shift assay indicated that a GATA binding site of the 15-LO-1 promoter region binds to GATA proteins present in both undifferentiated and, to a lesser extent, NaBT-treated (differentiated) Caco-2 cells. Moreover, that DNA binding shift band was disrupted after the addition of GATA-6 antibody in a supershift assay in the absence of NaBT, suggesting that GATA-6 is bound to the GATA binding site of the 15-LO-1 promoter in undifferentiated cells. In contrast, the addition of GATA-6 antibody did not affect the DNA binding ability in NaBT-induced differentiated cells. On the other hand, mutation of the GATA site of the 15-LO-1 promoter decreased the transactivation of the 15-LO-1 promoter as measured by luciferase activity in both FBS and NaBT cultured cells, indicating an unknown GATA binding protein to up-regulate 15-LO-1 expression. These implicate the GATA site at -240 of the proximal region of the 15-LO-1 promoter in the basic transcription of 15-LO-1 gene expression in Caco-2 cells, with GATA-6 acting to repress 15-LO-1 expression. PMID- 10675568 TI - The monomeric polypeptide comprises the functional flavanone 3beta-hydroxylase from Petunia hybrida. AB - Flavanone 3beta-hydroxylase catalyzes the Fe(II)/oxoglutarate-dependent hydroxylation of (2S)-flavanones to (2R,3R)-dihydroflavonols in the biosynthesis of flavonoids, catechins and anthocyanidins. The enzyme had been partially purified from Petunia hybrida and proposed to be active as a dimer of roughly 75 kDa in size. More recently, the Petunia 3beta-hydroxylase was cloned and shown to be encoded in a 41655 Da polypeptide. In order to characterize the molecular composition, the enzyme was expressed in a highly active state in Escherichia coli and purified to apparent homogeneity. Size exclusion chromatographies of the pure, recombinant enzyme revealed that this flavanone 3beta-hydroxylase exists in functional monomeric and oligomeric forms. Protein cross-linking experiments employing a specific homobifunctional sulfhydryl group reagent or the photochemical activation of tryptophan residues confirmed the tendency of the enzyme to aggregate to oligomeric complexes in solution. Thorough equilibrium sedimentation analyses, however, revealed a molecular mass of 39. 2+/-12 kDa for the recombinant flavanone 3beta-hydroxylase. The result implies that the monomeric polypeptide comprises the catalytically active flavanone 3beta hydroxylase of P. hybrida, which may readily associate in vivo with other proteins. PMID- 10675567 TI - A family of ubiquitin-like proteins binds the ATPase domain of Hsp70-like Stch. AB - We have isolated two human ubiquitin-like (UbL) proteins that bind to a short peptide within the ATPase domain of the Hsp70-like Stch protein. Chap1 is a duplicated homologue of the yeast Dsk2 gene that is required for transit through the G2/M phase of the cell cycle and expression of the human full-length cDNA restored viability and suppressed the G2/M arrest phenotype of dsk2Delta rad23Delta Saccharomyces cerevisiae mutants. Chap2 is a homologue for Xenopus scythe which is an essential component of reaper-induced apoptosis in egg extracts. While the N-terminal UbL domains were not essential for Stch binding, Chap1/Dsk2 contains a Sti1-like repeat sequence that is required for binding to Stch and is also conserved in the Hsp70 binding proteins, Hip and p60/Sti1/Hop. These findings extend the association between Hsp70 members and genes encoding UbL sequences and suggest a broader role for the Hsp70-like ATPase family in regulating cell cycle and cell death events. PMID- 10675569 TI - The polyomavirus major capsid protein VP1 interacts with the nuclear matrix regulatory protein YY1. AB - Polyomavirus reaches the nucleus in a still encapsidated form, and the viral genome is readily found in association with the nuclear matrix. This association is thought to be essential for viral replication. In order to identify the protein(s) involved in the virus-nuclear matrix interaction, we focused on the possible roles exerted by the multifunctional cellular nuclear matrix protein Yin Yang 1 (YY1) and by the viral major capsid protein VP1. In the present work we report on the in vivo association between YY1 and VP1. Using the yeast two-hybrid system we demonstrate that the VP1 and YY1 proteins physically interact through the D-E region of VP1 and the activation domain of YY1. PMID- 10675571 TI - Corrigendum to: conservation of the central proline-rich (PxxP) motifs of human immunodeficiency virus type 1 nef protein during the disease progression in two hemophiliac patients (FEBS 22727). PMID- 10675572 TI - Early CT changes of ischemia: limitations of what we know. PMID- 10675573 TI - Heparin and oral anticoagulants in the treatment of brain ischemia. AB - The use of heparin, heparin analogues, and oral anticoagulation in the treatment of brain ischemia remains controversial. We review the mechanism of action of the heparins and warfarin as well as the data regarding their efficacy and complications in patients with ischemic stroke and venous sinus thrombosis. None of these drugs have proven effective for all forms of cerebral ischemia. Further studies examining the efficacy of anticoagulants in different stroke subgroups will be necessary in order to clarify optimal clinical practice. PMID- 10675574 TI - Computed tomography findings in the first few hours of ischemic stroke: implications for the clinician. AB - In order to evaluate the clinical usefulness of emergency computed tomography (CT) in acute ischemic stroke, we assessed whether CT findings within the first few hours of stroke onset reliably predict type, site and size of the index infarction, and risk of death or disability. For this reason we reviewed clinical and CT findings in a cohort of unselected consecutive patients referred to the stroke unit of a large urban hospital because of a presumed ischemic stroke in the anterior circulation (AC), and submitted to CT within 5 h from onset. Out of 158 total patients, emergency CT revealed parenchymal changes compatible with AC focal ischemia in 77 (49%) and a hyperdense middle cerebral artery (MCA) in 41 (26%). Parenchymal changes and hyperdense MCA predicted an AC territorial infarction respectively in 97% of cases (95% C.I. 93% to 100%) and in 95% of cases (95% C.I. 88% to 100%). Site and size of early changes coincided with those of final lesions in 79% of patients with cortical changes and in 95% of patients with cortico-subcortical changes, but only in 37% of patients with initial subcortical changes, the remainder of whom developed a cortico-subcortical infarction. At logistic regression parenchymal changes were the only independent predictor of an AC territorial infarction. Negative predictive power, however, was only 40% (95% C. I. 29% to 51%) for parenchymal changes, and 35% for hyperdense MCA (95% C.I. 26% to 44%). The odds for death or disability at 1 month associated with parenchymal changes were thrice as high as with negative CT, even after adjustment for clinical severity on admission. These results indicate that CT scan adds significantly to the prediction of outcome made on clinical grounds. The frequent development of a territorial infarction in patients with initially negative CT and the subsequent recruitment of the cortex in those initially exhibiting only subcortical changes suggest that the transition from ischemia to infarction often occurs after the first five h following stroke. PMID- 10675575 TI - Medial temporal atrophy and memory impairment in early stage of Alzheimer's disease: an MRI volumetric and memory assessment study. AB - Memory impairment and medial temporal lobe (MTL) involvement are the earliest and most prominent features of Alzheimer's disease (AD). A psychological assessment of memory function and an evaluation of the morphological changes in MTL structures, as found in the mild form of AD, are important for early diagnosis as well as for understanding the pathophysiology of the disease. In the present study, we aimed to evaluate correlations in these psychoanatomical changes in terms of the stage of AD. We performed MRI-based volumetric measurements of the MTL structure and neuropsychological tests, using MMSE and the Wechsler memory scale-revised (WMS-R), on 27 elderly normal subjects and 46 probable AD patients, and then checked for possible correlations between the volumetric measurements and memory dysfunction. The severity of the AD patients' condition was assessed by CDR scale. Each MTL structure decreased in volume with increasing severity of AD. In very early AD, the reduction in the amygdala volume was pronounced, while the hippocampal volumes were relatively unchanged. Neuropsychological scores also declined with increasing severity of AD. Scores on the main WMS-R subsets examined (verbal memory, visual memory, and delayed recall) decreased significantly in the very mild group, as compared with normal controls. The WMS-R test scores correlated significantly with the amygdala volumes in normal control subjects and very mild AD patients. Our findings suggest that MRI-based amygdaloid volumetric measurement provides a sensitive marker, and that the degeneration of the amygdala may begin very early in the course of AD. PMID- 10675576 TI - Serial positron emission tomography (PET) in gliomatosis cerebri treated with radiotherapy: a case report. AB - Results of serial positron emission tomography (PET) in a biopsy-proven case of gliomatosis cerebri (GC) are reported. Computed tomography (CT) with and without contrast failed to detect focal abnormalities, but magnetic resonance (MR) revealed iso-intensity or low-intensity lesions in T1-weighted images and high intensity lesions in T2-weighted images. Lesions were seen in the left thalamus, right temporal lobe and claustrum, and pons. Radiotherapy remarkably improved clinical and imaging findings. Both before and shortly after radiotherapy, 11C methionine PET images showed hypermetabolism while 15O-water PET images showed a marked increase in cerebral blood flow in GC lesions. However, 6 months later PET images had remarkably improved, appearing nearly normal. PMID- 10675577 TI - Epstein-Barr virus nuclear antigen-1 (EBNA-1) associated oligoclonal bands in patients with multiple sclerosis. AB - Oligoclonal bands (OCBs) are frequently observed in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS), but the target antigens of these antibodies remain unknown. We used antigen specific immunoblotting to determine whether Epstein Barr virus nuclear antigen-1 (EBNA-1) was a target of the OCBs in the CSF of patients with MS. Antibody indices (AIs) were measured by ELISA and calculated by the formula of Reiber and Lange which includes correction factors for both breakdown of the blood brain barrier and intrathecal polyclonal IgG synthesis. A distinctive oligoclonal antigen specific banding pattern for EBNA-1 was observed in 5/15 MS patients, but 0/12 controls (P=0.037, Fisher's Exact Probability). AIs in this EBNA-l positive subgroup were extremely high, comparable with levels observed in viral CNS infections. In one patient with EBNA 1 specific OCBs, EBNA-1 and a peptide 'equivalent', p62, were able to absorb a component of the total IgG. Our results suggest that in a subset of MS patients, EBNA-1 may be a major target of selected OCBs. PMID- 10675578 TI - Changes in uptake of vitamin B(12) and trace metals in brains of mice treated with clioquinol. AB - Clioquinol is a hydroxyquinoline antibiotic that has been associated with severe side-effects in the CNS. The syndrome caused by clioquinol treatment, subacute myelo-optic neuropathy (SMON), is considered as one of the worst drug disasters of this century. The precise biochemical mechanism behind SMON is not fully understood. Clioquinol can form strong lipophilic chelates with divalent cations and therefore it has been speculated that the drug may disturb the retention of vitamin B(12) through chelation of Co(2+). In the present study, the tissue distribution and uptake capacity of [57Co]cyanocobalamin were estimated in mice treated with clioquinol or saline. The concentrations of some trace metals were also determined in brain tissue. Accumulation of vitamin B(12) in the brain and its concentration in blood were decreased by clioquinol treatment. The mean concentrations of several trace metals were also lowered in the brain while the concentration of cobalt in the brain was not affected, suggesting that clioquinol does not bind to the cobalt in vitamin B(12). Moreover, a significant decrease in the levels of S-adenosylmethionine (SAM) was observed in the brain after clioquinol treatment. This may be a consequence of decreased vitamin B(12) levels. From these results, it can be concluded that chronic treatment with clioquinol may alter the tissue homeostasis of vitamin B(12) in the brain. PMID- 10675579 TI - Marked increase of matrix metalloproteinase 9 in cerebrospinal fluid of patients with fungal or tuberculous meningoencephalitis. AB - Matrix metalloproteinases (MMPs) are believed to play an essential role in the breakdown of the extracellular matrix macromolecules in the blood-cerebrospinal fluid barrier and blood-brain barrier (BBB). In this study, the levels of MMP-2 and MMP-9 and their common tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) were measured in the cerebrospinal fluid (CSF) from patients with various meningitides including aseptic, fungal and tuberculous ones. MMP-9 production level in CSF was more increased in subacute meningitis including fungal and tuberculous meningitis than in acute aseptic meningitis and non-inflammatory neurological diseases (NIDs). Enhanced production of MMP-9 was associated with high proteolytic activity detected by gelatin zymography. The MMP-2 and TIMP-1 levels in CSF of subacute meningitis were also higher than those of NIDs. In contrast, the TIMP-2 levels in CSF of either acute aseptic or subacute meningitis were not up-regulated compared with those of NIDs. The central nervous system (CNS) complications (i.e. disturbance of consciousness, psychiatric symptoms, urinary disturbance, etc.) during the course of meningitis showed good correlation with the enhanced production of MMP-9 in CSF. Immunohistochemical studies in tuberculous meningitis demonstrated that the infiltrating mononuclear cells in the meninges were immunoreactive for both MMP-2 and MMP-9. However, the infiltrating mononuclear cells into CNS parenchyma had immunoreactivity for MMP 9, but not for MMP-2. Taken together, those data suggest that MMP-9 in CSF may be a useful marker of encephalitogenecity during the course of subacute meningitis. PMID- 10675580 TI - Aggregation of ubiquitin and a mutant ALS-linked SOD1 protein correlate with disease progression and fragmentation of the Golgi apparatus. AB - Transgenic mice that express the G93A mutation of human Cu,Zn superoxide dismutase (SOD1(G93A)), found in familial amyotrophic lateral sclerosis (FALS), showed clinical symptoms and histopathological changes of sporadic ALS, including fragmentation of the neuronal Golgi apparatus (GA). The finding of fragmented neuronal GA in asymptomatic mice, months before the onset of paralysis, suggests that the GA is an early target of the pathological processes causing neuronal degeneration. Transgenic mice expressing human SOD1(G93A) have aggregates of mutant protein and ubiquitin in neuronal and glial cytoplasm; they appeared first in the neuropil and later in the perikarya of motor neurons, where they were adjacent to fragmented GA. The aggregates of SOD1(G93A) appeared in neuronal perikarya of asymptomatic mice containing fragmented GA. The numbers of neurons with deposits of SOD1(G93A) and fragmented GA progressively increased with age. Immuno-electron microscopy using colloidal gold showed labeling of ubiquitin and SOD1 over 13 nm thick cytoplasmic filaments. Spinal cord extracts showed a 20 fold increase of SOD1(G93A) in transgenic mice compared to the wild-type protein in controls. The results suggest a causal relationship between the aggregation of mutant SOD1 and ubiquitin, fragmentation of the Golgi apparatus of motor neurons and neurodegeneration. PMID- 10675581 TI - The neuronal Golgi apparatus is fragmented in transgenic mice expressing a mutant human SOD1, but not in mice expressing the human NF-H gene. AB - Fragmentation of the Golgi apparatus (GA) of motor neurons was first described in sporadic amyotrophic lateral sclerosis (ALS) and later confirmed in transgenic mice expressing the G93A mutation of the gene encoding the enzyme Cu,Zn superoxide dismutase (SOD1(G93A)) found in some cases of familial ALS. In these transgenic mice, however, the fragmentation of the neuronal GA was associated with cytoplasmic and mitochondrial vacuoles not seen in ALS. The present new series of transgenic mice expressing 14-17 trans gene copies of SOD1(G93A), compared to 25 copies in the mice we studied previously, showed consistent fragmentation of the GA of spinal cord motor neurons, axonal swellings, Lewy-like body inclusions in neurons and glia, but none of the cytoplasmic or mitochondrial vacuoles originally reported. Thus, this animal model recapitulates the clinical and most neuropathological findings of sporadic ALS. Neurofilaments (NF) accumulate in axons and, less often, in neuronal perikarya in most cases of sporadic ALS and they have been implicated in its pathogenesis. In order to investigate whether fragmentation of the neuronal GA also occurs in association with accumulation of perikaryal NFs, we studied the organelle in transgenic mice expressing the heavy subunit of human neurofilaments (NF-H) which developed a motor neuronopathy resembling ALS. The neuronal GA of mice expressing NF-H, however, was intact despite massive accumulation of NFs in both perikarya and axons of motor neurons. In contrast, in transgenic mice expressing SOD1(G93A), the GA was fragmented despite the absence of accumulation of perikaryal NFs. These findings suggest that, in transgenic mice with neuronopathies caused by the expression of mutant SOD1(G93A) or the human NF-H, the GA and the perikaryal NFs are independently involved in the pathogenesis. The evidence suggests that the GA plays a central role in the pathogenesis of the vast majority of sporadic ALS and in FALS with SOD1 mutations. PMID- 10675582 TI - The effect of oral and intravenous methylprednisolone treatment on subsequent relapse rate in multiple sclerosis. AB - We investigated the effect of oral and intravenous methylprednisolone treatment on subsequent relapse rate in patients with multiple sclerosis. Following a double blind trial designed to compare the effect of oral and intravenous methylprednisolone treatment on promoting recovery from acute relapses of multiple sclerosis, 80 patients were followed for two years with six-monthly assessments during which all subsequent relapses were recorded. The annual relapse rate was slightly higher in the oral compared with the intravenous methylprednisolone-treated patients (1.06 vs. 0.78), but the adjusted difference between the two groups was not statistically significant (0.18; 95% CI -0.19 to 0.55, P=0.3). The time to onset and the severity of the first relapse after treatment, the number of relapse free patients at the end of the follow-up period, and the severity of the relapses during the follow-up period were similar in the two groups. This trial did not show a statistically significant difference in relapse rate during the first two years following oral compared with intravenous methylprednisolone treatment. PMID- 10675583 TI - Neuromyotonia in association with essential thrombocythemia. PMID- 10675584 TI - Hyaluronidases of Gram-positive bacteria. AB - Bacterial hyaluronidases, enzymes capable of breaking down hyaluronate, are produced by a number of pathogenic Gram-positive bacteria that initiate infections at the skin or mucosal surfaces. Since reports of the hyaluronidases first appeared, there have been numerous suggestions as to the role of the enzyme in the disease process. Unlike some of the other more well studied virulence factors, much of the information on the role of hyaluronidase is speculative, with little or no data to substantiate proposed roles. Over the last 5 years, a number of these enzymes from Gram-positive organisms have been cloned, and the nucleotide sequence determined. Phylogenetic analysis, using the deduced amino acid sequences of the Gram-positive hyaluronidases, suggests a relatedness among some of the enzymes. Molecular advances may lead to a more thorough understanding of the role of hyaluronidases in bacterial physiology and pathogenesis. PMID- 10675585 TI - Chlorobiphenyl-desleucyl-vancomycin inhibits the transglycosylation process required for peptidoglycan synthesis in bacteria in the absence of dipeptide binding. AB - Novel glycopeptide analogs are known that have activity on vancomycin resistant enterococci despite the fact that the primary site for drug interaction, D-ala-D ala, is replaced with D-ala-D-lactate. The mechanism of action of these compounds may involve dimerization and/or membrane binding, thus enhancing interaction with D-ala-D-lactate, or a direct interaction with the transglycosylase enzymes involved in peptidoglycan polymerization. We evaluated the ability of vancomycin (V), desleucyl-vancomycin (desleucyl-V), chlorobiphenyl-vancomycin (CBP-V), and chlorobiphenyl-desleucyl-vancomycin (CBP-desleucyl-V) to inhibit (a) peptidoglycan synthesis in vitro using UDP-muramyl-pentapeptide and UDP-muramyl tetrapeptide substrates and (b) growth and peptidoglycan synthesis in vancomycin resistant enterococci. Compared to V or CBP-V, CBP-desleucyl-V retained equivalent potency in these assays, whereas desleucyl-V was inactive. In addition, CBP-desleucyl-V caused accumulation of N-acetylglucosamine-beta-1, 4 MurNAc-pentapeptide-pyrophosphoryl-undecaprenol (lipid II). These data show that CBP-desleucyl-V inhibits peptidoglycan synthesis at the transglycosylation stage in the absence of binding to dipeptide. PMID- 10675586 TI - Regulation of aflatoxin production by Ca(2+)/calmodulin-dependent protein phosphorylation and dephosphorylation. AB - To elucidate Ca(2+)-mediated regulation of aflatoxin production, the status of Ca(2+)/calmodulin-dependent protein phosphorylation and dephosphorylation was investigated employing toxigenic and non-toxigenic strains of Aspergillus parasiticus. Incubation of cytoplasmic extracts with [gamma-(32)P]ATP followed by SDS-PAGE and autoradiography revealed total absence of protein phosphorylation during periods corresponding to aflatoxin production in the toxigenic strain (NRRL 2999). In contrast, protein phosphorylation was unaffected in the non toxigenic strain (SRRC 255). Aflatoxin production in the toxigenic strain was also accompanied by enhanced (26-fold) activity of calcineurin (calmodulin dependent protein phosphatase 2B) concomitant with a lowered (6-fold) activity of calmodulin-dependent protein kinase. In addition, the in vitro activity of Ca(2+)/calmodulin-dependent protein kinase was susceptible to dose-dependent inhibition by aflatoxin. Since calcineurin remains active in the absence of phosphorylation by calmodulin-dependent protein kinase, it is suggested that calcineurin-mediated dephosphorylation of regulatory enzymes ensures continued production of aflatoxins. PMID- 10675587 TI - Conserved tyr residues determine functions of Alicyclobacillus acidocaldarius squalene-hopene cyclase. AB - The catalytic cavity of Alicyclobacillus acidocaldarius squalene-hopene cyclase is mainly lined by aromatic amino acids. In recombinant cyclases, three out of four tyrosine residues (Y) have been mutated to phenylalanine residues (F). The mutant cyclases Y495F and Y612F had less activity than the wild-type cyclase, but a wild-type product pattern. Mutant Y609F had wild-type activity but a drastically altered product pattern with hopene and significant amounts of bicyclic alpha-polypodatetraene and different tetracyclic triterpenes (dammaradienes and eupha-7,24-diene). The experiments demonstrated that Y495 and Y612 may be involved in the initiation of the cyclization reaction and Y609 in the stabilization and/or positioning of the intermediate carbocations. PMID- 10675588 TI - Separation of NADH-fumarate reductase and succinate dehydrogenase activities in Trypanosoma cruzi. AB - A recent review suggested that the activity of NADH-fumarate reductase from trypanosomatids could be catalyzed by succinate dehydrogenase working in reverse (Tielens and van Hellemond, Parasitol. Today 14, 265-271, 1999). The results reported in this study demonstrate that the two activities can easily be separated without any loss in either activity, suggesting that fumarate reductase and succinate dehydrogenase are separate enzymes. PMID- 10675589 TI - Expression of green fluorescent protein in Lactococcus lactis. AB - The gfp gene from Aequorea victoria, encoding the green fluorescent protein (GFP) has been expressed in Lactococcus lactis subsp. lactis biovar cremoris MG1363, upon construction and introduction of plasmid pLS1GFP into this host. GFP was monitored in living cells during growth to evaluate its use in molecular and physiological studies. Quantification of the levels of GFP expressed by cultures was feasible by fluorescence spectroscopy. Phase-contrast and fluorescence microscopy allowed us to distinguish, in mixed cultures, lactococcal cells expressing GFP. Our results indicate that GFP can be used as a reporter in L. lactis. PMID- 10675590 TI - Relationships between the anti-HIV V(3)-derived peptide SPC(3) and lymphocyte membrane properties involved in virus entry: SPC(3) interferes with CXCR(4). AB - SPC(3) is a multiple antigen peptide derived from the V(3) loop of human immunodeficiency virus (HIV) envelope (Env). It exerts a potent anti-HIV activity whereas it alters neither Env expression nor binding to CD(4). Here, SPC(3) binding characteristics, its subsequent intracellular fate and the fact that it inhibited SDF(1)alpha binding to the lymphocyte surface provided strong arguments to conclude that it exerts its anti-HIV activity through interference with the CXCR(4) coreceptor. In contrast, it interferes with none of the other major surface proteins and mechanisms involving V(3) and implicated in infection, as shown here. This work identifies the target mechanism of SPC(3). PMID- 10675591 TI - Identification and molecular analysis of superoxide dismutase isoforms in Helicobacter pylori. AB - Three electromorphs of iron superoxide dismutase (FeSOD) were identified among 29 Helicobacter pylori isolates by native gel electrophoresis and activity staining. The electromorphs designated isoforms A, B, and C are characterized by slow, intermediate and fast electrophoretic migration, respectively, which was not observed under denaturing conditions. The isoforms were not associated with virulence determinants and with the outcome of disease. Sequence analysis of the sodB gene in strains producing different FeSOD isoforms and comparison of deduced protein sequences revealed that differences in the electric migration behavior are associated with exchange of charged amino acids, suggesting that faster migration is caused by a more negative total charge of the proteins. Electrophoretic migration of native FeSOD was not influenced by changes in the iron cofactor concentration, oxidative stress, and different media, indicating that FeSOD isoforms represent stable strain-specific markers. PMID- 10675593 TI - In vitro transcription system using reconstituted RNA polymerase (Esigma(70), Esigma(H), Esigma(E) and Esigma(S)) of Pseudomonas aeruginosa. AB - We have developed an in vitro transcription system for Pseudomonas aeruginosa genes, using RNA polymerase (RNAP) holoenzyme reconstituted with purified sigma protein and RNAP core enzyme. The RNAP core enzyme was directly purified from P. aeruginosa PAO1 cells. The sigma factors of P. aeruginosa (sigma(70), sigma(H), sigma(E) and sigma(S)) were prepared in a hexa-histidine tagged form, which were expressed in Escherichia coli and purified using a HisTrap Chelating column. The RNAP holoenzyme reconstituted from core enzyme with each sigma factor recognized correctly each of the cognate promoters. This system will be useful for the promoter analysis of many genes in P. aeruginosa. PMID- 10675592 TI - The Bacillus subtilis ctaB paralogue, yjdK, can complement the heme A synthesis deficiency of a CtaB-deficient mutant. AB - Heme A is a prosthetic group in many respiratory oxidases. It is synthesised from heme B (protoheme IX) with heme O as an intermediate. In Bacillus subtilis two genes required for heme A synthesis, ctaA and ctaB, have been identified. CtaB is the heme O synthase and CtaA is involved in the conversion of heme O to heme A. A ctaB paralogue, yjdK, has been identified through the B. subtilis genome sequencing project. In this study we show that when carried on a low copy number plasmid, the yjdK gene can complement a ctaB deletion mutant with respect to heme A synthesis. Our results indicate that YjdK has heme O synthase activity. We therefore suggest that yjdK be renamed as ctaO. PMID- 10675594 TI - Aldolases of the DhnA family: a possible solution to the problem of pentose and hexose biosynthesis in archaea. AB - Sequence analysis of the recently identified class I aldolase of Escherichia coli (dhnA gene product) helped to identify its homologs in Chlamydia trachomatis, Chlamydiophyla pneumoniae and in each of the completely sequenced archaeal genomes. Iterative database searches revealed sequence similarities between the DhnA-family enzymes, deoxyribose phosphate aldolases and bacterial (class II) fructose bisphosphate aldolases and allowed prediction of similar three dimensional structures (TIM-barrel fold) in all these enzymes. The Schiff base forming lysyl residues of DhnA and deoxyribose phosphate aldolase are conserved in all members of the DhnA and deoxyribose phosphate aldolase families, indicating that these enzymes share common features with both class I and class II aldolases. The DhnA-family enzymes are predicted to possess an aldolase activity and to play a critical role in sugar biosynthesis in archaea. PMID- 10675595 TI - Mannanase Man26A from Cellulomonas fimi has a mannan-binding module. AB - A modular mannanase (Man26A) from the bacterium Cellulomonas fimi contains a mannan-binding module (Man26Abm) that binds to soluble but not to insoluble mannans. Man26Abm does not bind to cellulose, chitin or xylan. The K(d) for binding of Man26Abm to locust bean gum (LBG) is approximately 0.2 microM. Man26A is the first mannanase reported to contain a mannan-binding module. PMID- 10675596 TI - Plasmid curing from an acidophilic bacterium of the genus Acidocella. AB - Preservation of the acidophilic heterotroph, Acidocella sp. strain GS19h, at 4 degrees C in stab culture eliminated all indigenous plasmids from this bacterium. Growth at 42 degrees C initially caused changes in the plasmid profile followed by total elimination of plasmids after 10 cycles of growth. Concomitant to this loss of all plasmids, the cured derivatives became sensitive to CdSO(4) and ZnSO(4), and the MIC value of the salts dropped from 1 M for each in the case of parental strain to 2 mM and 5 mM, respectively, suggesting plasmid-mediated inheritance of metal resistance in this bacterium. The cured derivatives could not utilise lactose, indicating this metabolic activity to be plasmid-associated in this strain. PMID- 10675597 TI - Detection of genes for membrane-bound nitrate reductase in nitrate-respiring bacteria and in community DNA. AB - A nested PCR primed by four degenerate oligonucleotides was developed for the specific amplification of sequences from the narG gene encoding the membrane bound nitrate reductase. This approach was used to amplify fragments of the narG gene from five Pseudomonas species previously shown to be able to express the membrane-bound nitrate reductase and from community DNA extracted from a freshwater sediment. Amino acid sequences encoded by the narG fragments were compared to one another, and to the corresponding regions of related enzymes. This comparison indicates that the amplification protocols are specific for their intended targets. Sequences amplified from community DNA were tightly clustered, which may indicate a degree of homogeneity in the sediment community. The PCR primers and amplification protocols described will be useful in future studies of nitrate respiring populations. PMID- 10675598 TI - Cloning, over-expression and purification of Pseudomonas aeruginosa murC encoding uridine diphosphate N-acetylmuramate: L-alanine ligase. AB - We cloned and sequenced the murC gene from Pseudomonas aeruginosa encoding a protein of 53 kDa. Multiple alignments with 20 MurC peptide sequences from different bacteria confirmed the presence of highly conserved regions having sequence identities ranging from 22-97% including conserved motifs for ATP binding and the active site of the enzyme. Genetic complementation was done in Escherichia coli (murCts) suppressing the lethal phenotype. The murC gene was subcloned into the expression vector pET30a and overexpressed in E. coli BL21(lambdaDE3). Three PCR cloning strategies were used to obtain the three recombinant plasmids for expression of the native MurC, MurC His-tagged at N terminal and at C-terminal, respectively. MurC His-tagged at C-terminal was chosen for large scale production and protein purification in the soluble form. The purification was done in a single chromatographic step on an affinity nickel column and obtained in mg quantities at 95% homogeneity. MurC protein was used to produce monoclonal antibodies for epitope mapping and for assay development in high throughput screenings. Detailed studies of MurC and other genes of the bacterial cell cycle will provide the reagents and strain constructs for high throughput screening and for design of novel antibacterials. PMID- 10675599 TI - The Yersinia high-pathogenicity island is present in different members of the family Enterobacteriaceae. AB - A pathogenicity island termed high-pathogenicity island (HPI) is present in pathogenic Yersinia. This 35 to 45 kb island carries genes involved in synthesis, regulation and transport of the siderophore yersiniabactin. Recently, the HPI was also detected in various strains of Escherichia coli. In this study, the distribution of the HPI in the family Enterobacteriaceae was investigated. Among the 67 isolates pertaining to 18 genera and 52 species tested, nine (13.4%) harbored the island. These isolates were three E. coli, one Citrobacter diversus and five Klebsiella of various species (Klebsiella pneumoniae, Klebsiella rhinoscleromatis, Klebsiella ozaenae, Klebsiella planticola, and Klebsiella oxytoca). As in Yersinia sp., all nine isolates synthesized the HPI-encoded iron repressible proteins HMWP1 and HMWP2. In the K. oxytoca strain, the right-end portion of the HPI was deleted, whereas the entire core region of the island was present in the eight other enterobacteria strains analyzed. In most of these isolates, the HPI was bordered by an asn tRNA locus, as in Yersinia sp. This report thus demonstrates the spread of the HPI among various members of the family Enterobacteriaceae. PMID- 10675600 TI - Bacterial triterpenoids of the hopane series as biomarkers for the chemotaxonomy of Burkholderia, Pseudomonas and Ralstonia spp. AB - Hopanoid fingerprints allowed to differentiate bacteria formerly connected to the genus Pseudomonas. Whereas all strains related to Pseudomonas and Ralstonia were devoid of any detectable hopanoid, these pentacyclic triterpenoids were found in the Burkholderia species and in related soil isolates, which contained as main hopanoid a bacteriohopanetetrol carbapseudopentose ether, accompanied by significant amounts of its novel Delta(6) unsaturated homologue. Unsaturated hopanoids represent an extremely rare feature in soil bacteria and the only known indication for a catabolism of this pentacyclic carbon skeleton in bacteria. PMID- 10675601 TI - Selection of Pycnoporus cinnabarinus strains for laccase production. AB - A comparison of Pycnoporus cinnabarinus strains for laccase production was carried out. A dikaryotic strain, I-937 strain, producing a high level of laccase (9500 U l(-1)) was selected. The study of the life cycle in vitro of this dikaryotic strain led to isolation of monokaryons. Forty-eight monokaryotic strains were isolated and screened for laccase production. One of these strains, ss3, produced a higher level of laccase than the parental strain I-937. The maximum production reached 29000 U l(-1) in medium supplemented with ferulic acid. PMID- 10675602 TI - A non-stop antisense reading frame in the grp78 gene of Neurospora crassa is homologous to the Achlya klebsiana NAD-gdh gene but is not being transcribed. AB - A long non-stop reading frame exists on the antisense strand of the grp78 gene (cDNA and genomic DNA) of Neurospora crassa. Computer analysis revealed a strong similarity of the putative antisense protein to the 10th exon of the NAD dependent glutamate dehydrogenase gene (NAD-gdh) of Achlya klebsiana, which is itself located on the complementary strand of a transcribed hsc70 gene homologue. In Neurospora, no grp78 antisense mRNA was detected by Northern blot and reverse transcription-coupled polymerase chain reaction analyses, indicating that this long reading frame is not being transcribed. Hypotheses for the presence of such unexpressed non-stop reading frames are discussed. PMID- 10675603 TI - Copper accumulation by sulfate-reducing bacterial biofilms. AB - Sulfate-reducing bacterial biofilms were grown in continuous culture. When exposed to medium containing 20 or 200 microM Cu, biofilms accumulated Cu. Energy dispersive X-ray analysis (EDXA) showed that accumulation of Cu occurred in the form of sulfides while EDXA mapping of Cu and S in biofilm sections indicated that they were not uniformly distributed but located in the surface of the biofilm. While the polymer content of biofilm exposed to 20 microM Cu did not appear to increase relative to control Cu-free biofilms, biofilms exposed to 200 microM Cu accumulated carbohydrate and smaller amounts of protein throughout the incubation period. The mechanism of uptake, therefore, appeared to be precipitation of Cu sulfides at the biofilm surface or in the liquid phase followed by entrapment of precipitated Cu sulfide by the exopolymer-enhanced biofilm. PMID- 10675604 TI - Interleukin-4-deficient BALB/c mice develop an enhanced Th1-like response but control cardiac inflammation following Borrelia burgdorferi infection. AB - Interleukin-4 has been reported to critically modulate Borrelia burgdorferi infection and Lyme arthritis in experimental murine models. To determine the in vivo role of IL-4 in controlling Lyme carditis, we compared immunological responses and the severity of cardiac inflammation in wild-type BALB/c (IL-4 +/+) and IL-4 deficient BALB/c (IL-4 -/-) mice infected with B. burgdorferi by tick bite. At day 15 and 30 post-infection IL-4 -/- mice produced significantly greater titers of spirochete-specific IgG2a than the wild-type IL-4 +/+ mice, which produced significantly more spirochete-specific IgG1. Following in vitro antigenic stimulation with B. burgdorferi antigen, splenocytes from infected IL-4 -/- and IL-4 +/+ mice displayed similar magnitudes of proliferative responses at day 15 and 30 post-infection. At day 30 antigen-stimulated splenocytes from infected IL-4 -/- mice, however, produced significantly more IFN-gamma than those derived from similarly infected IL-4 +/+ mice, suggesting that Th1-influenced responses predominated in IL-4 -/- mice. Moreover, inflamed hearts from IL-4 -/- mice displayed higher levels of IFN-gamma and TNF-alpha transcripts as compared to IL-4 +/+ mice. At both time points antigen-stimulated splenocytes from IL-4 +/+ and IL-4 -/- mice produced significant amounts of IL-10 but those from IL-4 +/+ mice produced either no or little IL-4. Histopathology demonstrated typical Lyme carditis in both IL-4 +/+ and IL-4 -/- mice at day 15 and day 30. Although Borrelia-infected IL-4 -/- mice developed a more severe carditis on day 30, the carditis resolved by day 50, as it did in IL4 +/+ mice. These results indicate that although IL-4 may help limit the severity of Lyme carditis, its absence does not preclude resolution of cardiac lesions. PMID- 10675605 TI - Regulation of the plasma membrane potential in Pneumocystis carinii. AB - Many protists use a H(+) gradient across the plasma membrane, the proton motive force, to drive nutrient uptake. This force is generated in part by the plasma membrane potential (DeltaPsi). We investigated the regulation of the DeltaPsi in Pneumocystis carinii using the potentiometric fluorescent dye bisoxonol. The steady state DeltaPsi in a buffer containing Na(+) and K(+) (standard buffer) was found to be -78+/-8 mV. In the absence of Na(+) and K(+) (NMG buffer) or Cl(-) (gluconate buffer), DeltaPsi was not significantly changed suggesting that cation and anion conductances do not play a significant role in the regulation of DeltaPsi in P. carinii. The DeltaPsi was also not affected by inhibitors of the Na(+)/K(+)-ATPase, ouabain (1 mM), and the K(+)/H(+)-ATPase, omeprazole (1 mM). In contrast, inhibitors of the plasma membrane H(+)-ATPase, dicyclohexylcarbodiimide (100 microM), N-ethylmaleimide (100 microM) and diethylstilbestrol (25 microM), significantly depolarized the DeltaPsi to -43+/ 7, -56+/-5 and -40+/-12 mV, respectively. The data support that the plasma membrane H(+)-ATPase plays a significant role in the regulation of DeltaPsi in P. carinii. PMID- 10675606 TI - Intact Cryptosporidium parvum oocysts isolated after in vitro excystation are infectious to neonatal mice. AB - In vitro excystation is often used as a measure of viability of encysted protozoan parasites. Parasites that do not excyst in vitro are assumed to be non viable and non-infectious, whereas those that do excyst are assumed viable. To test the validity of these assumptions, Cryptosporidium parvum oocysts were excysted in vitro using two different excystation protocols, and the non-excysted intact oocysts were isolated using flow cytometry. Non-excysted sorted oocysts readily infected neonatal CD-1 mice. Increasing the duration of the excystation assays from 1 h to 3 h resulted in a higher percent of excysted oocysts, but the remaining non-excysted parasites were still capable of infecting neonatal CD-1 mice. Our results suggest that in vitro excystation is not an accurate measure of the viability or infectious potential of C. parvum oocysts. PMID- 10675607 TI - The Drosophila primo locus encodes two low-molecular-weight tyrosine phosphatases. AB - The fine modulation of tyrosine phosphorylation by protein tyrosine phosphatases and protein tyrosine kinases is a key regulatory mechanism for many cell signaling pathways active during development. In a screen for genes with interesting expression patterns in the developing Drosophila pupal retina, we identified a novel pair of protein tyrosine phosphatases that exhibit an expression pattern suggesting a role in multiple steps of Drosophila neurogenesis. Together, these phosphatases define the primo locus. Their sequence is approx. 50% identical to each other and to low-molecular-weight protein tyrosine phosphatases (LMW-PTPs) identified in other species. Little is understood of the biological role of LMW-PTPs, and the powerful tools available in Drosophila should provide important insight into their role in signaling and development. PMID- 10675608 TI - Structure and chromosomal localization of the rat salivary Psp and Smgb genes. AB - SMGB and PSP are among the most abundant products of the immature acinar cells in developing rat parotid and submandibular glands and are also products of the sublingual gland serous demilunes. Previous analysis of Smgb and Psp cDNA clones demonstrated a high degree of sequence similarity between the signal peptide encoding and 3' untranslated regions of these transcripts, although the secreted proteins themselves are more divergent. The current study reports the upstream sequences, genomic organization and localization of the Psp and Smgb genes. Both structural genes contain nine exons and are present at 3q41-3q42, where they are arranged in tandem and separated by 21kb. In addition to the previously observed sequence similarity, Psp and Smgb are highly homologous throughout exon 1 and at 365 of 600bp immediately upstream of the transcription start site. These findings indicate that the Psp and Smgb genes arose by tandem duplication and divergence. The similar neonatal submandibular and parotid gland expression patterns observed for these genes are likely to be due to closely conserved or shared enhancer(s). PMID- 10675609 TI - Chain reaction cloning: a one-step method for directional ligation of multiple DNA fragments. AB - A novel DNA assembly method, chain reaction cloning (CRC), is described. CRC enables the ordered assembly of multiple DNA fragments in a single step. The power of the technique was demonstrated by the directed in vitro assembly of a plasmid comprised of six DNA fragments from a pool of 12 available fragments. The odds of obtaining the correct plasmid clone in a single step, using conventional techniques, is less than 1 in 191000000. Using CRC, the desired plasmid was recovered at a frequency of one in two. Ligation is no longer the rate limiting step in cloning, and limitless possibilities exist for the reconstruction of complex genomes. PMID- 10675610 TI - Cloning of a human gene closely related to the genes coding for the c-myc single strand binding proteins. AB - Southwestern screening of human fibroblast cDNAs with an upstream element of the alpha2(I) collagen promoter (Box 5A) has led to the identification of a novel gene product (RBMS3). RBMS3 contains two pairs of RNA binding motifs and is very closely related to the structure of the c-myc gene single-strand binding proteins (MSSPs). MSSPs are believed to regulate DNA replication, transcription, apopotosis and cell cycle progression by interacting with the C-MYC protein. Consonant with this postulate, RBMS3 binds in vitro to the minus strand of Box 5A and transactivates transcription in the chimeric GAL4 hybrid system. However, the RBMS3 protein mostly localizes to the cytoplasm of transfected cells, in addition to binding strongly in vitro to synthetic poly-U and poly-A oligoribonucleotides. Finally, overexpression in transfected fibroblasts of RBMS3 with and without a nuclear localization signal has no effect on Box 5A-driven transcription. The results thus exclude RBMS3 involvement in the transcriptional regulation of COL1A2 and strongly suggest a cytoplasmic function of this new member of the MSSP family. As part of the initial characterization of RBMS3 we have also established that the gene resides on human chromosome 3p23-p24 and is widely expressed in the embryo and in the adult organism. PMID- 10675611 TI - Nob1p, a new essential protein, associates with the 26S proteasome of growing saccharomyces cerevisiae cells. AB - Nob1p, which interacts with Nin1p/Rpn12, a subunit of the 19S regulatory particle (RP) of the yeast 26S proteasome, has been identified by two-hybrid screening. NOB1 was found to be an essential gene, encoding a protein of 459 amino acid residues. Nob1p was detected in growing cells but not in cells in the stationary phase. During the transition to the stationary phase, Nob1p was degraded, at least in part, by the 26S proteasome. Nob1p was found only in proteasomal fractions in a glycerol gradient centrifugation profile and immuno-coprecipitated with Rpt1, which is an ATPase component of the yeast proteasomes. These results suggest that association of Nob1p with the proteasomes is essential for the function of the proteasomes in growing cells. PMID- 10675612 TI - Crossover hot-spot instigator (Chi) sequences in Escherichia coli occupy distinct recombination/transcription islands. AB - Crossover hot-spot instigator (Chi) sequences (5'-GCTGGTGG-3') are orientation dependent, strand-specific sequences implicated in RecA-mediated DNA recombination. In Escherichia coli and Haemophilus influenzae Chi and Chi-like sequences preferentially locate to approx. 1kb recombination 'islands' in the mRNA-synonymous strands of open reading frames (ORFs). Since mRNA-synonymous strands follow Szybalski's transcription direction rule in being G-rich, and the average ORF is about 1kb, then, on this basis alone, Chi sequences are seen to reside in 1kb G-rich 'islands'. However, RecA preferentially binds GT-rich sequences, suggesting that genomic context might potentiate Chi action. Consistent with this, we report for E. coli that 1kb sequence windows with Chi near their centres are a distinct subset of total 1kb windows, the mRNA synonymous strands being preferentially enriched in both G and T. Chi function might be particularly important for bacteria that survive high temperature and radiation. These often exist in habitats where recombination with E. coli DNA would be unlikely, so canonical Chi sequences might not confer a selective disadvantage in this respect. In general, Chi sequences are not more frequent in thermophilic bacteria and Deinococcus radiodurans, than in E. coli and other mesophilic bacteria. Only two of five thermophilic bacteria examined showed preferential location of Chi sequences to mRNA-synonymous strands. In the thermophile Methanococcus jannaschii, windows containing the canonical Chi sequence do not form a distinct subset. We suggest that in thermophilic bacteria and D. radiodurans the Chi function may be achieved by sequences that differ from the canonical Chi sequence, or that the number of these sequences is sufficient, or that the Chi function is unnecessary. PMID- 10675613 TI - Sequence, genomic organization and functional expression of the murine tRNA specific adenosine deaminase ADAT1. AB - We have recently identified the first mammalian tRNA-specific adenosine deaminase human ADAT1, a member of the ADAR family of RNA editing enzymes. This protein is responsible for the first step of the unique A(37) to m(1)I(37) modification in eukaryotic tRNA(Ala). Here, we present the genomic structure of murine ADAT1 and the functional expression of mADAT1 cDNA. In mouse, as well as in human, ADAT1 is expressed from a single copy gene. The coding region of the mADAT1 gene is spread over nine exons, covering approximately 30kb of genomic DNA and encodes a protein of 499 amino acids. Overall, mADAT1 shares 81% nucleotide homology and 87.5% protein homology with the human ortholog. The recombinant mouse protein is active specifically and with a high efficiency on human tRNA(Ala) in vitro. Its genomic organization is compared to the structures of the sequence-related, pre-mRNA specific adenosine deaminases ADAR1 and ADAR2. PMID- 10675614 TI - Conservation of sequence and function of the pag-3 genes from C. elegans and C. briggsae. AB - The Caenorhabditis briggsae homologue of the Caenorhabditis elegans pag-3 gene was cloned and sequenced. When transformed into a C. elegans pag-3 mutant, the C. briggsae pag-3 gene rescued the pag-3 reverse kinker and lethargic phenotypes. The C. elegans pag-3 gene fused to lacZ was expressed in the same pattern in C. elegans and C. briggsae. Unlike many gene homologues compared between C. elegans and C. briggsae, extensive sequence conservation was found in the non-coding regions upstream of the pag-3 exons, in several of the introns and in the downstream non-coding region. Furthermore, the splice acceptor and splice donor sites were conserved, and the size of the introns and exons was surprisingly similar. The predicted protein sequence of C. briggsae PAG-3 was 85% identical to the protein sequence of C. elegans PAG-3. Because so much of the non-coding region of pag-3 was conserved, the control of pag-3 may be quite complex, involving the binding of many trans-acting factors. These results suggest the evolutionary conservation of the pag-3 gene sequence, its expression and function. PMID- 10675615 TI - Genomic organization and transcriptional analysis of the human genes coding for caveolin-1 and caveolin-2. AB - Caveolin-1 and caveolin-2 are related proteins involved in the biogenesis of caveolae. The corresponding genes in humans (CAV and CAV2, respectively), have been mapped to a common locus in chromosome 7q31.1, and are possible candidates for the tumor suppressor gene postulated in this region. Here, we show that CAV and CAV2 are independent transcriptional units lying in the same orientation, with CAV2 centromeric and about 17kb upstream to CAV. The two genes have similar tissue expression patterns. Alternative termination/polyadenylation generates two CAV2 mRNAs. Multiple transcriptional start sites spanning 35bp upstream from the CAV2 ATG are detected by 5' RACE, consistent with a TATA-less promoter predicted by sequence analysis. The CAV2 promoter region contains two SRE-like boxes resembling those described in the CAV promoter and proposed to link transcription to intracellular cholesterol levels. However, exogenous sterols had only minor effects on CAV and CAV2 RNA levels in HeLa cells, suggesting that SREBPs are not sufficient to regulate caveolin transcription. PMID- 10675616 TI - An isozyme of the NADP-malic enzyme of a CAM plant, Aloe arborescens, with variation on conservative amino acid residues. AB - In Aloe arborescens, an obligate CAM plant, Western analysis detected three major isoforms of NADP-malic enzyme (NADP-ME), 72kDa with a pI of 6.0, 65kDa with a pI of 5.6 and 65kDa with a pI of 5.5. Among them, the 65kDa protein with a pI of 5.5 was leaf-specific, and the 65kDa protein with a pI of 5.6 was found only in roots, whereas the 72kDa protein was uniformly detected in both organs. Activity staining indicated enzyme activity of both 65kDa NADP-MEs but little activity of the 72kDa protein. A cDNA clone encoding a leaf-abundant NADP-ME, AME1, was isolated. Deduced amino acid sequence of AME1 showed a high degree of homology to known NADP-MEs, but it was also found that AME1 contained substitutions on five conservative amino acid residues, some of which have been predicted to be important for their enzyme activity. Transgenic rice carrying the aloe AME1 gene efficiently produced an additional 65kDa protein with a pI of 5.5 as an active NADP-ME. These results indicate that AME1 corresponds to the leaf-specific 65kDa NADP-ME, which may be involved in CAM photosynthesis. It was also shown that substitutions of these conservative amino acid residues identified in AME1 still allowed it to give enzyme activity. PMID- 10675617 TI - Structural characterisation of the mouse nuclear oxysterol receptor genes LXRalpha and LXRbeta. AB - Oxysterols are important regulatory molecules of diverse biological processes such as cholesterol homeostasis, bile acid synthesis and apoptosis. Recent findings led to the suggestion that some of these functions are mediated by the nuclear receptors LXRalpha and LXRbeta owing to their potential to bind a group of naturally occurring oxysterols as their ligands. In this report, we compare the genomic structure and the promoter regions of the two mouse LXR genes. In addition, we show evidence for the presence of a processed, but truncated LXRbeta pseudogene in the mouse genome. RACE-PCR on mouse liver cDNA demonstrates the presence of more than one defined transcription initiation site for both genes. The LXRalpha and LXRbeta promoter regions are GC-rich and contain a number of putative Sp1 binding sites but lack obvious TATA and CAAT boxes. A database search revealed several sequence motifs in the LXR promoter regions that resemble known transcription factor binding sites. Most striking is the identification of one potential NFkappaB and seven potential Ets-protein binding sites in the LXRbeta promoter, suggesting an important role for this receptor in the haematopoietic/immune system. PMID- 10675618 TI - Alfalfa (Medicago sativa) carbamoylphosphate synthetase gene structure records the deep lineage of plants. AB - Given the central role of carbamoylphosphate synthetases in pyrimidine and arginine metabolism in all living organisms, the absence of fundamental information regarding plant CPSase genes is a striking omission [Lawson et al., Mol. Biol. Evol. 13 (1996) 970-977; van den Hoff et al., J. Mol. Evol. 41 (1995) 813-832]. Whereas CPSase gene architecture and aa sequence have proven to be useful characters in establishing ancient and modern genetic affinities, phylogenetic analysis cannot be completed without the inclusion of plant CPSases. We describe the first isolation by molecular cloning of a plant CPSase gene (CPAII) derived from alfalfa (Medicago sativa). DNA sequence analysis reveals a proteobacterial architecture, namely closely linked carA and carB coding domains separated by a short intergenic region, and transcribed as a polycistronic mRNA. CPAII encodes the amino acid residues that typify a CPSase type II enzyme. In addition, an ancient internal duplication has been retained in the plant carB sequence. Partial nucleotide sequencing of additional clones reveals that the alfalfa genome contains multiple CPSase II gene copies which may be tissue specific in their expression. It appears that with respect to CPSase genes, CPAII resembles the carAB gene of bacteria, and may have preserved much of this ancient gene structure in the alfalfa genome. PMID- 10675619 TI - Isolation and genomic analysis of the human surf-6 gene: a member of the Surfeit locus. AB - The human Surfeit locus contains at least six tightly clustered genes (Surf-1 to Surf-6) of which five (Surf-1 to Surf-5) have been characterised and found not to share any sequence homology. The organisation and juxtaposition of the Surfeit genes are conserved between human and mouse. The Surf-6 gene that encodes a novel nucleolar-matrix protein with nucleic-acid binding properties has been characterised in mouse. In this work, we have isolated and analysed the human Surf-6 homologue and determined its genomic organisation in the Surfeit locus. The human Surf-6 gene has five exons spread over a distance of 4.3kb and has features of a housekeeping gene being ubiquitously expressed, having its 5' end located within a CpG rich island and lacking a canonical TATA box. The intragenic region between the 3' end of the Surf-5 gene and the 5' end of the Surf-6 gene is 3.2kb and contains a pseudogene of the ribosomal protein gene rpL21. The putative human Surf-6 protein is 361 amino acids long and includes motifs found in both the mouse and fish Surf-6 homologues, which may underlie the functions of Surf-6. Three amino acid polymorphisms have been detected at codons 163, 175 and 311 by SSCP analysis. PMID- 10675620 TI - Role of the ruvB gene in homologous and homeologous recombination in Rhizobium etli. AB - The Rhizobium etli ruvA and ruvB genes were cloned through a PCR-based approach, using degenerate primers matching conserved sectors in the amino acid sequences of RuvB from eight bacterial species. Comparative analysis of the predicted polypeptides for RuvA and RuvB of R. etli showed highly conserved blocks with the corresponding homologs in other bacteria; RuvB depicts characteristic motifs for DNA helicases (ATP-binding and DEXH-box motifs). An R. etli ruvB::loxP Sp mutant was constructed by interposon mutagenesis. This mutant was highly sensitive to DNA-damaging agents, such as methyl methanesulfonate and nitrofurantoin, implying a deficiency in DNA repair. Homologous and homeologous conjugational recombination was reduced almost tenfold in the ruvB::loxP Sp mutant; a recombination defect was also observed in assays employing recombination between small plasmids, albeit at a smaller magnitude. Although the ruvA and ruvB genes are contiguous in R. etli, complementation studies suggest that they are expressed independently. PMID- 10675621 TI - An effective family shuffling method using single-stranded DNA. AB - Family shuffling, which is one of the most powerful techniques for in vitro protein evolution, always involves the problem of reassembling the gene fragments into parental gene sequences, because such a process prevents the formation of chimeric sequences. In order to improve the efficiency of hybrid formation in family shuffling, single-stranded DNAs (ssDNAs) were used as templates. The ssDNAs of two catechol 2,3-dioxygenase genes, nahH and xylE, were prepared, the xylE strand being complementary to the nahH strand. When these ssDNAs were digested by DNase I and reassembled, chimeric genes were obtained at a rate of 14%, which was much higher than the rate of less than 1% obtained by shuffling with double-stranded DNAs. Chimeric catechol 2,3-dioxygenases that were more thermally stable than the parental enzymes, XylE and NahH, were obtained by this ssDNA-based DNA shuffling. PMID- 10675622 TI - A comparison of the pectate lyase genes, pel-1 and pel-2, of Colletotrichum gloeosporioides f.sp. malvae and the relationship between their expression in culture and during necrotrophic infection. AB - Extracellular pectic lyase and polygalacturonase activities of Colletotrichum gloeosporioides f.sp. malvae were detected in broths containing mallow cell wall extract, pectin or glucose as the carbon source. The initial pH of the broth as well as the carbon source had major influences on pectinase enzyme activities. In the host, only pectic lyase activity was detected, which began at the end of the biotrophic phase and increased in the necrotrophic phase of infection. Two full length pectate lyase cDNAs, pel-1 and pel-2, were cloned from the fungus. Both genes showed similar patterns of expression when the fungus was grown in mallow cell-wall extract and pectin medium, and the only major difference in expression in culture was that only pel-2 was expressed in glucose broth. Expression of pel 1 and pel-2 was also affected by the initial pH of the medium. Expression of pel 2, but not pel-1, was detected during infection of the host, round-leaved mallow, Malva pusilla. Transcripts of pel-2 were first detectable during the necrotrophic phase of infection approx. 24h after the first detection of pectic lyase enzyme activity. A comparison of expression of pel-1 and pel-2 in culture and in planta with other pectinase genes of C. gloeosporioides f.sp. malvae, as well as with other plant pathogenic fungi, indicates that expression during necrotrophic infection correlates with the ability to be expressed in media containing glucose. PMID- 10675623 TI - Molecular cloning, characterization and expression of a novel retinal clusterin like protein cDNA. AB - A novel gene expressed predominantly in retina, but detected at a conspicuously lower level in retina of canine progressive rod cone degeneration (prcd), has been identified by suppression subtractive hybridization and retinal cDNA library screening. The characterized region of cDNA of the novel gene includes 1017 nucleotides of coding sequence predicted to encode a protein of 338 amino acids (M(r) 39389), 791 nucleotides of 5'-untranslated region (UTR), and 300 nucleotides of 3'-UTR including the poly(A)(+) tail. Multiple transcripts were detected in retina by Northern blot analysis, and a lower level of expression was observed in brain and liver by RT-PCR. The transcript appears to be developmentally regulated with a burst in gene expression at a time period (34 postnatal days) that coincides with the photoreceptor differentiation phase of retinal development. The deduced amino acid sequence from the cDNA of the novel gene has 24% identity and 48% similarity with the multifunctional glycoprotein clusterin. Hence, the putative gene product from the novel transcript has been named clusterin-like protein 1 (CLUL1). The human homologue of CLUL1 cDNA has 84 and 70% identity at the level of nucleotides and amino acids, respectively, with the characterized canine cDNA. The presence of a stretch of 128 amino acids in the putative human CLUL1, not detected in canine CLUL1, suggests alternate splicing events. An STS database search revealed that the human homologue of CLUL1 maps to chromosome 18p, a location not yet reported to harbor an RP locus. Tissue-specific expression of CLUL1 in retina, and its lower abundance in different forms of PRA suggest that this novel gene may represent an as-yet unidentified locus for a retinal disorder. PMID- 10675624 TI - Exon-intron organization of the human gp130 gene. AB - The exon-intron organization and sequences of the exon-intron boundaries of the human gp130 transmembrane receptor gene have been determined using genomic DNAs as samples. The gp130 gene comprises 17 exons and 16 introns. The positions of the exon-intron boundaries show good correlation to the functional/homology regions of gp130. Exons 3-17 code for the gp130 protein, and each subdomain of the receptor is encoded by a set of exons. The coding potential of exons and the intron phasing of the human gp130 gene conform to the patterns observed previously for other cytokine receptor genes. This supports the notions that the gp130 gene evolved from the same ancestral gene that gave rise to other members of the cytokine receptor family. PMID- 10675625 TI - cDNA cloning, characterization, expression and recombinant protein production of leukemia inhibitory factor (LIF) from the marsupial, the brushtail possum (Trichosurus vulpecula). AB - A reverse transcription technique using RNA templates combined with polymerase chain reaction (RT-PCR) was used to clone the cDNA fragment encoding the amino acid sequence of mature LIF protein of the marsupial, the brushtail possum, Trichosurus vulpecula. A PCR product with expected size, of 546bp, and termed tvLIF, was obtained using cDNA reverse-transcribed from total RNA isolated from possum uterus. A genomic DNA fragment (about 650bp) between the specified primers was also amplified, indicating the similarity in structure and organization of this gene and LIF genes from studied eutherian species, although the full-length of its cDNA and genomic DNA needs to be further clarified. The deduced amino acid sequence of tvLIF shows a high level of sequence identity and similar molecular characteristics to eutherian LIF, which suggests similar biological actions of this molecule in this marsupial. Because the expression of LIF gene in other mammalian species has been found to be at very low levels and its transcripts cannot be detected by Northern hybridization analysis, the expression pattern of tvLIF in adult tissues and reproductive tracts during early development was investigated using the RT-PCR technique. Resultant products of the RT-PCR were further analyzed by Southern hybridization using tvLIF as a probe. tvLIF transcripts were detected in most of the adult tissues and in the reproductive tracts of pregnant females. These results lend support to the idea that LIF contributes to the maintenance of pregnancy in this marsupial. PMID- 10675626 TI - Characterization of a Rab11 homologue in Trypanosoma cruzi. AB - Vesicle trafficking between organelles occurs through fusion of donor and specific acceptor membranes. This process is highly regulated and ensures proper direction in sorting and packaging of a number of molecules in eukaryotic cells. Monomeric GTPases of the Rab family play a pivotal role in the control of membrane fusion and vesicle traffic. In this paper, we characterize a Trypanosoma cruzi Rab 11 homologue (TcRab11) that shares at, the amino acid level, 40% similarity with human rab11, Arabdopsis thaliana rab11 and yeast rab11 homologue genes. Western blot analysis, using a polyclonal rabbit antiserum raised against a synthetic peptide derived from the COOH-terminus of predicted the TcRab11 protein, reacted to a 26kDa protein. In immunofluorescence assays, TcRab 11, was shown to be expressed in epimastigote and amastigote forms, but it was absent in trypomastigotes. Interestingly, the TcRab11 product seems to be located at the reservosome complex, a site of active endocytosis and vesicle fusion present only in the epimastigote stage. Therefore, TcRab11 may represent the first molecular marker of this peculiar organelle. PMID- 10675627 TI - Combinatorial gene expression using multiple episomal vectors. AB - Episomal vectors offer a powerful alternative to integrative recombination for transgene expression in mammalian cells. In this study, various combinations of G protein-coupled receptors (GPCRs) and the G protein subunit G(i2)alpha, were stably expressed from separate episomal vectors in 293-EBNA (293E) cells. Each episome did not adversely affect the others, as gauged by episomal copy number, steady-state mRNA levels and the presence of functional receptors and G protein. Cell lines expressing genes from multiple autonomously replicating vectors were stable just two weeks after transfection, and remained stable in continuous culture for at least 5months. Co-expression of supplementary G(i2)alpha with receptor amplifies the magnitude of signal transduction thereby permitting the development of more sensitive high throughput functional assays. Given these results, combinatorial transfection is the strategy of choice for generating stable cell lines expressing multiple genes for the study of signal-transduction pathways or the evaluation of receptor ligands. PMID- 10675628 TI - RLR1 (THO2), required for expressing lacZ fusions in yeast, is conserved from yeast to humans and is a suppressor of SIN4. AB - We isolated a mutation (rlr1-1; required for lacZ RNA) in the Saccharomyces cerevisiae (Sc) RLR1 gene as a suppressor of sin4, a component of the Mediator subcomplex of the RNA polymerase II holoenzyme and a determinant of chromatin structure. RLR1 encodes a deduced protein found also in fission yeast, nematode worms, and humans. The presence of these orthologs suggests that Rlr1 family members comprise a class of putative KEKE motif-containing proteins, characteristic of certain chaperones as well as regulators and subunits of the mammalian 20S proteasome. A role for RLR1 (THO2) in transcription appears to occur at a step subsequent to transcription initiation (see also Piruat, J.I. and Aguilera, A., 1998. EMBO J. 17, 4859-4872); Sc genes fused to the reporter gene lacZ were expressed at a very low level, while the corresponding native chromosomal genes were expressed at approximately normal levels in rlr1 mutants. Our studies show that rlr1 mutations cause a wide range of growth defects in addition to their novel affect on lacZ. PMID- 10675629 TI - An ascidian glycine-rich RNA binding protein is not induced by temperature stress but is expressed under a genetic program during embryogenesis. AB - We have cloned a putative ascidian glycine-rich RNA binding protein gene, CiGRP1. Its maternal transcript and protein are stored in the unfertilized egg. They are gradually decreased during the first few rounds of cleavage. The CiGRP1 zygotic transcript and protein start to accumulate at the gastrula stage. The CiGRP1 transcript is expressed in the brain precursor and mesenchyme precursor cells of the gastrula and the neurula stage, and the brain and mesenchyme cells of the tailbud stage embryo. The CiGRP1 protein is found in all nuclei and in the cytoplasm of brain and mesenchyme cells. Although many glycine-rich RNA binding protein homologs of plants and vertebrates are cold-inducible, CiGRP1 cannot be induced by cold shock or heat shock at the transcriptional and translational levels during embryogenesis. The temporal expression pattern and the tissue restricted expression pattern of CiGRP1 suggest that it has important roles in the very early stage of development and in the brain and the mesenchyme tissue specification. PMID- 10675630 TI - Endolymphatic hydrops induced by chronic administration of vasopressin. AB - Recently, many lines of evidence have supported the possibilities that vasopressin (VP) is closely linked to the formation of endolymphatic hydrops in Meniere's disease. In the present study, it was examined whether or not the chronic administration of VP might induce endolymphatic hydrops. For this purpose, histological studies and VP radioimmunoassay were independently performed in 20 and 40 guinea pigs, respectively. The degree of hydrops was quantitatively assessed by the increase ratio (IR) of the scala media area in the mid-modiolar sections of the cochlea. The IR was defined by the following equation: 100x(A-B)/B (A: the cross-sectional area of the bulging scala media; B: the no-bulging scala media, enclosed by an idealized straight Reissner's membrane). VP was administered at the rates of 200 microU/kg/min, 400 microU/kg/min and 1000 microU/kg/min for 1 week via the osmotic mini-pump. The IR of the total of the apical, second, third and basal turns (means+/-S.D.s) were 4.4+/-0.7, 10.4+/-1.8, 17.4+/-7.9 (n=10 ears, each) in respective doses of VP. Comparing with that of the control animals (5.2+/-1.7, n=10 ears), the area increased significantly in the VP dosage of 400 and 1000 microU/kg/min (Bonferroni's method, P<0.05). Plasma VP concentrations produced by the VP administration in these dosages were 2.2+/-0.4, 3.5+/-0.8 and 14.0+/-3.9 (n=10, each) pg/ml. Although 3.5 pg/ml is the upper limit of plasma VP concentration in normal human subjects, 14.0 pg/ml was almost the same concentration as those observed in the acute phase of Meniere's disease (Takeda et al., 1995). Therefore, the formation of endolymphatic hydrops in cases of Meniere's disease might be caused by high concentrations of plasma VP. PMID- 10675631 TI - Modulation of spontaneous activity by acetylcholine receptors in the rat dorsal cochlear nucleus in vivo. AB - In vitro studies have implicated muscarinic acetylcholine receptors (mAChRs) in the modulation of spontaneous activity (SA) of neurons in the rat dorsal cochlear nucleus (DCN) (Chen et al., 1994,1998). Early studies suggest that cholinergic pathways also modulate SA in vivo, but these effects have not been investigated pharmacologically. The purpose of the present study was to determine whether multiunit SA can be modulated in vivo by application of cholinergic agents to the surface of the DCN. Sprague Dawley rats were used in the current experiment. The influence of cholinergic activation on SA was tested by applying carbachol (5-500 microM) to the DCN surface while recording multiunit SA at a depth of 250 microm. Out of a total of 32 sites tested, all but 2 (94%) showed well-defined responses to carbachol, characterized by suppression, activation or a combination of both (two-component responses). The most common responses were pure suppression and suppression accompanied by transient activation. Both the proportion of sites showing suppressive responses and the magnitude of suppression averaged across sites increased with dose. Although the proportion of sites showing pure activation in response to carbachol decreased with dose, there was no clear trend in the magnitude of activation with dose. The suppressive responses to high doses of carbachol were blocked by pre-application of atropine. These results extend previous work by suggesting that muscarinic receptors play an important role in the modulation of SA in vivo. PMID- 10675632 TI - Neonatal sensorineural hearing loss affects neurone size in cat auditory midbrain. AB - We examined the effect of a neonatal sensorineural hearing loss on the soma area of neurones in the central nucleus of the inferior colliculus (ICC) in adult cats to evaluate the role of auditory experience on neuronal atrophy within the auditory midbrain. Three groups of animals were used: bilaterally deafened, unilaterally deafened and normal hearing controls. Soma area measurements were made from the laminated central and medial divisions of the ICC of eight deafened and two normal hearing cats. A small but significant reduction in soma area was evident for bilaterally deafened animals compared with normal hearing controls (P<0.05, Dunnett's test). In contrast, there was no significant difference in mean soma area between normal hearing and unilaterally deafened animals (P0.05) irrespective of whether the ICC examined was ipsi- or contralateral to the deafened ear. These results demonstrate that the reduction in soma area of auditory brainstem neurones reported following a sensorineural hearing loss is also evident at the level of the auditory midbrain. PMID- 10675633 TI - Calcium binding proteins and the AMPA glutamate receptor subunits in gerbil cochlear nucleus. AB - The alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) glutamate receptors mediate fast excitatory synaptic transmission in the central nervous system. The GluR2 subunit confers calcium impermeability to AMPA receptors. Various calcium binding proteins play a role in calcium regulation within the neurons. This study sought to identify possible relationships between calcium binding proteins and glutamate receptor subunits, especially GluR2, in gerbil cochlear nucleus neurons. Our immunohistochemical observations reveal no particular correlation between GluR2 and calbindin; all the cell types show labeling for all the antibodies studied except calretinin. There was coincidence of strong GluR4 and strong parvalbumin staining in octopus cells, although calbindin was also present in these cells. This study suggests a possible relationship between parvalbumin and predominantly GluR4 containing receptors, even when calbindin is present. The absence of a strong inverse correlation between the presence of ionotropic AMPA receptor subunit GluR2 and calbindin suggests a more significant role of non-AMPA ionotropic glutamate receptors or other voltage-gated channels in the regulation of calcium in the neurons of cochlear nucleus. Alternatively, more detailed analysis of receptor composition at particular synapses and the subcellular localization of specific calcium binding proteins may be required. PMID- 10675634 TI - Cisplatin-induced hearing loss: influence of the mode of drug administration in the guinea pig. AB - Cisplatin (8 mg/kg) was given intravenously to guinea pigs either as a 15 s bolus injection (25 animals) or as a 1 h infusion (28 animals). To determine the influence of the mode of cisplatin administration and pharmacokinetics on the ototoxic side-effect, the concentrations of cisplatin and the biotransformation product monoaquated cisplatin were determined in blood ultrafiltrate using liquid chromatography with post-column derivatization. Ototoxic effect was evaluated as difference in pre- and 96 h post-exposure auditory brainstem response (ABR) threshold. The cisplatin peak concentration was considerably higher, 19.2+/-2.4 microg/ml, in the bolus injection group than in the infusion group, 6.7+/-0.5 microg/ml (mean+/-S.E.M.). The area under the blood ultrafiltrate concentration time curve (AUC) for cisplatin was slightly greater in the infusion group, 442+/ 26 microg/ml/min, than in the bolus injection group, 340+/-5 microg/ml/min. For monoaqua cisplatin, the AUC was not different between the groups (bolus injection: 30.8+/-1. 5 microg/ml/min, infusion: 34.1+/-3.3 microg/ml/min). A significant ototoxic effect was observed in both groups at 20 and 12.5 kHz, but there was no difference between the groups in the extent of threshold shift. The interindividual variability in susceptibility to ABR threshold shift was far greater than the variability in pharmacokinetics, suggesting that other factors are more important in determining the degree of hearing loss. PMID- 10675635 TI - Electrical stimulation of the auditory nerve. III. Response initiation sites and temporal fine structure. AB - Latency, temporal dispersion and input-output characteristics of auditory nerve fiber responses to electrical pulse trains in normal and chronically deafened cat ears were classified and tentatively associated with sites where activity is initiated. Spikes occurred in one or more of four discrete time ranges whose endpoints overlapped partially. A responses had latencies <0.44 ms, exhibited asymptotic temporal dispersion of 8-12 micros and possessed an average dynamic range of 1.2 dB for 200 pulses/s (pps) pulse trains. They likely originated from central processes of spiral ganglion cells. B(1) and B(2) responses (0.45-0.9 ms, 25-40 micros, 1.9 dB) likely stemmed from activity at myelinated and unmyelinated peripheral processes, respectively. C100 micros) likely originated from direct stimulation of inner hair cells, and D8 dB) arose from propagating traveling waves possibly caused by electrically induced motion of outer hair cells. C and D responses were recorded only in acoustically responsive ears. Mean latencies of spikes in all time ranges usually decreased with intensity, and activity at two or even three discrete latencies was often observed in the same spike train. Latency shifts from one discrete time range to another often occurred as intensity increased. Some shifts could be attributed to responses to the opposite polarity phase of the biphasic pulse. In these cases, temporal dispersion and dynamic range were approximately equal for activity at each latency. A second type of latency shift was also often observed, in which responses at each latency exhibited dissimilar temporal dispersion and dynamic range. This behavior was attributed to a centralward shift in the spike initiation site and it occurred for monophasic as well as biphasic signals. Several fibers exhibited dual latency activity with a 40-90 micros time difference between response peaks. This may have stemmed from spike initiation at nodes on either side of the cell body. Increasing the stimulus pulse rate to 800-1000 pps produced small increases in temporal dispersion and proportionate increases in asymptotic discharge rate and dynamic range, but thresholds did not improve and slopes of rate-intensity functions (in spikes/s/dB) did not change. Responses to high-rate stimuli also exhibited discrete latency increases when discharge rates exceeded 300-400 spikes/s. Spike by spike latencies in these cases depended strongly on the discharge history. Implications for high-rate speech processing strategies are discussed. PMID- 10675636 TI - Differential expression of voltage-gated potassium channel genes in auditory nuclei of the mouse brainstem. AB - Voltage-gated potassium (Kv) channels may play an important role in the encoding of auditory information. Towards understanding the roles of Shaker and Shaw-like channels in this process, we examine here the expression of Kv1.1, Kv1.2, Kv3.1, and Kv3.3 in the central auditory nuclei of the mouse using quantitative in situ hybridization techniques. We establish rank order for each channel's expression in each region, finding that the medial nucleus of the trapezoid body shows the highest signal for each of the four channel genes. In other auditory nuclei differential expression is found among and between members of both Shaker and Shaw subfamilies. Of particular interest is the stark contrast between high level expression of Kv1.1 and very low level expression of Kv3.1 in the octopus cell area of the cochlear nucleus and in the lateral superior olivary nucleus. These unique expression patterns suggest that Kv channel gene expression is regulated to allow brainstem auditory neurons to transmit temporally patterned signals with high fidelity. In instances where specific cell types can be tentatively identified, we discuss the possible contribution made by these channel genes to the physiological properties of those neurons. PMID- 10675637 TI - Lipopolysaccharide-induced expression of inducible nitric oxide synthase in the guinea pig organ of Corti. AB - The purpose of the investigation was to ascertain whether inoculation of bacterial lipopolysaccharide (LPS) into the cochlea of the guinea pig could elicit formation of inducible nitric oxide synthase (iNOS). Immunohistochemical study revealed that immunoreactivity to iNOS was seen below outer hair cells representing nerve fibers and synaptic nerve endings. iNOS-staining could also be observed in phalangeal dendrites of Deiter's cells pointing to the cuticular membrane, Hensen's cells and on stria vascularis 48 h after inoculation with LPS. Immunohistochemical investigation with a specific anti-nitrotyrosine antibody also revealed intense immunoreactivity identical to that of iNOS, suggesting formation of peroxynitrite in the organ of Corti by the reaction of NO with O(2)( ). On the basis of these findings, it can be concluded that NO together with O(2)(-), which form the more reactive peroxynitrite, are the most important pathogenic agents in LPS-induced damage of cochlea in the guinea pig. PMID- 10675638 TI - Evidence for multiple DPOAE components based upon group delay of the 2f(1)-f(2) distortion in the gerbil. AB - The cochlear delay of the 2f(1)-f(2) distortion product otoacoustic emission (DPOAE) was measured using the phase gradient method. With a constant f(2) and swept f(1), the resulting phase change of 2f(1)-f(2) was used to calculate the group delay for f(2) frequencies from 1 to 60 kHz. For f(2) frequencies between 2 and 60 kHz, the group delays were between 2.2 and 0.11 ms and continuously decreased for increasing f(2) and for increasing primary stimulus levels. For f(2) frequencies below 2 kHz, the group delay decreased to around 1 ms and was largely independent of stimulus level. The ratio curves resulting from the f(1) sweeps for high frequencies (f(2)16 kHz) displayed the typical mammalian shape with a peak in the level of 2f(1)-f(2) for a larger primary frequency separation (f(2)/f(1)1.15) and decreasing 2f(1)-f(2) level for smaller primary separation. In addition to this typical level maximum, for f(2) frequencies from about 1.8 to 16 kHz, the ratio curves displayed a second component in the form of an increase in the level of 2f(1)-f(2) for small primary separation at higher primary levels (level of f(2)30 dB SPL). For f(2) frequencies below 1.8 kHz, only the second component and no typical ratio peak as for higher f(2) could be observed and the associated group delay was always close to 0.8 ms. Several possible causes for this behavior are discussed, including different modes of DPOAE generation and modulation as well as changes in the nature of mechanical processing from base to apex in the gerbil cochlea. To evaluate the relative sensitivity of non-linear cochlear mechanics, an iso-distortion threshold curve was constructed from acoustical growth functions of the 2f(1)-f(2) DPOAE at optimum primary separation, by plotting the level of f(2) sufficient to evoke a distortion of -10 dB SPL as a function of f(2)2.5 kHz but failed to reflect the sensitivity for lower frequencies. This may be a consequence of more linear frequency processing in the apex. PMID- 10675639 TI - Activation of medial olivocochlear efferent system in humans: influence of stimulus bandwidth. AB - The activity of the medial olivocochlear bundle (MOCB) can be studied in humans through variations in the level of evoked otoacoustic emissions (EOAEs) elicited by contralateral acoustic stimuli (CAS). The present study sought to investigate how the activity of the MOC system at a given frequency, as measured through the contralateral suppression of tone-pip EOAEs, depends on the bandwidth of the contralateral stimulus. EOAEs were recorded in 155 normal-hearing subjects, successively with and without contralateral stimuli whose bandwidth, center frequency and level were systematically varied. We showed a clear dependence of contralateral EOAE suppression on bandwidth demonstrating increased suppression with increased bandwidth over about two octaves around the center frequency of the noise. This effect was obtained irrespective of whether contralateral noise energy was kept constant independently of bandwidth or not, which indicates a role of bandwidth per se in contralateral EOAE suppression. Results are interpreted in terms of a simple model of MOCB activation mechanisms including peripheral bandpass filtering, within-channel compression and across-channel spatial summation by the afferent paths. Complementary experiments suggested a greater effectiveness of increases in bandwidth on the upper than on the lower side and of frequency components akin to or remote from the test frequency than of intermediate bands. Finally, these results were complemented by detailed spectrum analyses of the EOAE level variations induced by the different noises, which revealed that whilst noise components close to or remote from the center frequency generally attenuated EOAE level, intermediate components could in some cases lead to a relative increase in EOAE level. These results can further be explained by assuming different positive and negative weights on the inputs to the spatial summation process depending on their position relative to the center frequency. PMID- 10675640 TI - Glycine induced calcium concentration changes in vestibular type I sensory cells. AB - Glutamate is the neurotransmitter of the synapse between vestibular type I hair cells and the afferent nerve calyx. This calyx may also be involved in local feedback, which may modify sensory cell activity via N-methyl-D-aspartate (NMDA) receptors. Glycine is the co-agonist of glutamate in NMDA receptor activation. Both agents have been detected by immunocytochemistry in the nerve calyx. Glutamate and NMDA stimulations cause changes in the intracellular calcium concentration ([Ca(2+)](i)) of isolated type I sensory cells. We investigated the effect of glycine stimulation on [Ca(2+)](i) in guinea pig type I sensory cells by spectrofluorimetry with fura-2. Glycine application to isolated type I sensory cells induced a rapid and transient increase in [Ca(2+)](i). The fluorescence ratio increased by 55% above the resting level. The peak was reached in 9 s and the return to basal level took about 20 s. A specific antagonist of the glycine site on NMDA receptors, 7-chlorokynurenate (10 microM), decreased the calcium response to glycine by 60%. Glycine may activate NMDA receptors. Glycine may also activate the strychnine-sensitive glycine receptor-gated channel. Strychnine (50 microM) decreased the calcium response to glycine by 60%. Thus, glycine probably induces calcium concentration changes in type I vestibular sensory cells via NMDA receptors and/or glycine receptors. PMID- 10675641 TI - Classification and culture of spiral ligament fibrocytes from mice. AB - In this study, we established an immunocytochemical strategy to classify the fibrocytes of the murine spiral ligament (SL), and SL cultures were characterized. Similar to those in other mammals, three different types of fibrocytes were identified. Type I fibrocytes, which are found lateral to the stria vascularis, showed positive immunoreactivity for caldesmon and S-100 protein and were not stained for sodium-potassium-adenosinetriphosphatase (Na-K ATPase). Type II fibrocytes are located lateral to the spiral prominence epithelium and suprastrial region, and they were distinguishable by their positive staining for Na-K-ATPase. Type III fibrocytes, which are found adjacent to bone in the inferior region of the SL, contained caldesmon but not S-100 or Na K-ATPase. Secondary cultures from the SL were positive for caldesmon and S-100 and negative for Na-K-ATPase, suggesting that these cells were type I fibrocytes. The present immunocytochemical approach was useful for the classification of murine fibrocyte cultures, and these cultures may benefit future immunological studies of the inner ear because mice have been well characterized immunologically. PMID- 10675642 TI - Cochlear findings in the white spotting (Ws) rat. AB - White spotting (Ws) rats possess a c-kit gene mutation at the W locus, resulting in a variety of characteristics including a lack of intermediate cells of the stria vascularis. The present study employs a light microscope (LM), scanning (SEM) and transmission electron microscopes (TEM), diaminobenzidine (DAB) staining techniques and auditory brainstem response (ABR) to investigate the structure and function of the cochlea in 26 homozygous Ws/Ws rats aged 1-6 months. A slight thinning of the stria vascularis and moderate elevation of ABR threshold were about the only defects noted in 1 month animals, while older animals displayed various defects that tended to worsen with age. At 3 months LM revealed pigment granules in the basal turn of most animals, with a loss of pigmentation in the upper turns. The stria vascularis and organ of Corti tended to be well preserved in the lower, pigmented portion, while the upper, unpigmented portion showed severe strial degeneration and some outer hair cell loss. DAB staining revealed a well developed strial capillary net throughout the pigmented portion of the cochlea, with severe degradation in the unpigmented apical portion. ABR thresholds were slightly elevated over 1 month values. At 6 months great differences in degeneration were noted between right and left ears of the same animal. PMID- 10675643 TI - The influence of noise on blood flow in the basilar artery (BA) - measurements with transcranial color-coded duplex sonography (TCCD). AB - Acoustic stimuli are being reported as a cause of changes in resistance in the basilar artery (BA). It was the aim of this study to investigate this effect under standardized conditions dependent upon the intensity of the evoking stimulus. Twenty healthy subjects with normal hearing (male/female 14/6; mean age 26.4 years) were exposed to 'pink noise' for periods of 2 min at 75, 85 and 95 dB(A). Parallel to this, the Doppler spectrum of the BA and both the Pourcelot resistance index and the Gosling pulsatility index were measured by means of transcranial color-coded Doppler sonography. In comparison with the base value (at rest) a significant increase in resistance was noted during noise exposure. The noise-induced resistance changes could be interpreted as a consequence of changes in activity of the various centers of the auditory pathway and cerebral function. Further animal experiments may prove the connection between BA blood flow and resistance and their changes depending on different acoustic stimuli or different hearing pathophysiology. PMID- 10675645 TI - Detecting coherent and incoherent frequency modulation. AB - Three experiments investigated whether or not the auditory system contains a neural mechanism that is sensitive to differences in the pattern of frequency modulation imposed on widely separated carriers. Experiment 1 measured the discrimination between an unmodulated two-tone complex and one in which either coherent or incoherent frequency modulation was imposed on the two carrier frequencies. These two frequencies were either 1100+1925 Hz or 1100+2000 Hz, and the stimuli were presented against a pink-noise background. The method was based on that used in experiments by Furukawa and Moore (1996), which were previously interpreted as providing evidence in favour of a mechanism sensitive to FM coherence. Discrimination was sometimes better for coherent than for incoherent FM, as reported by Furukawa and Moore, but only for four out of the eight listeners tested. The remaining experiments excluded those subjects who had shown no effect of FM coherence in experiment 1. Experiment 2 showed that detection of a static shift on the carrier frequencies of the two components was better when the carriers were shifted in the same, compared to the opposite, direction. This difference occurred regardless of whether the carriers were modulated coherently, incoherently, or were unmodulated. The experiment also showed that performance was better when the 1100-Hz carrier was shifted down and the 1925-Hz carrier was shifted up, compared to when the 1100-Hz carrier shifted up and the 1925-Hz carrier shifted down. Experiment 3 showed that this difference also applied to dynamic changes: detection of quasi-linear frequency sweeps (0.5 cycles of sinusoidal FM) was better when the higher component glided up and the lower component glided down than vice versa. In the former condition, performance was as good as with same-direction sweeps. It is concluded that the effects observed in experiment 1 and by Furukawa and Moore result from the processing of a global percept arising from the perceptual fusion of the two carriers, and do not represent an across-frequency mechanism sensitive to FM coherence. In addition, it is argued that experiments 2 and 3 demonstrate the existence of perceptual asymmetries in hearing. PMID- 10675644 TI - Hyperactivity in the dorsal cochlear nucleus after intense sound exposure and its resemblance to tone-evoked activity: a physiological model for tinnitus. AB - Intense tone exposure induces increased spontaneous activity (hyperactivity) in the dorsal cochlear nucleus (DCN) of hamsters. This increase may represent an important neural correlate of noise-induced tinnitus, a condition in which sound, typically of very high pitch, is perceived in the absence of a corresponding acoustic stimulus. Since high pitch sounds are thought to be represented in central auditory structures by the place of activation across the tonotopic array; it is therefore possible that the high pitch of noise-induced tinnitus occurs because intense sound exposure induces a tonotopic distribution of chronic hyperactivity in the DCN similar to that normally evoked only under conditions of high frequency stimulation. To investigate this possibility we compared this tone induced hyperactivity with the activity evoked in normal animals by presentation of a tone. This comparison revealed that the activity in the DCN of animals which had been exposed to an intense 10 kHz tone 1 month previously showed a striking similarity to the activity in the DCN of normal animals during presentation of low to moderate level tonal stimuli of the same frequency. In both test conditions similar patterns were seen in the topographic distribution of the increased activity along the tonotopic axis. The magnitude of hyperactivity in exposed animals was similar to the evoked activity in the normal DCN responding to a stimulus at a level of 20 dB SL. These results suggest that the altered DCN following intense tone exposure behaves physiologically as though it is responding to a tone in the absence of a corresponding acoustic stimulus. The relevance of these findings to noise-induced tinnitus and their implications for understanding its underlying mechanisms are discussed. PMID- 10675646 TI - Middle ear influence on otoacoustic emissions. I: noninvasive investigation of the human transmission apparatus and comparison with model results. AB - Evoked otoacoustic emissions (EOAEs) are generated within the cochlea in response to external sounds, and they can be acoustically detected in the external auditory meatus after backward propagation through the middle ear. In addition to being used to probe the cochlear mechanisms, they are expected to be sensitive to minute changes in middle ear impedance. Systematic measurements of the changes in the vectorial components of EOAEs were carried out after various manipulations of the human middle ear in order to characterize the influence of stiffness and inertia of the stapes and tympanic-membrane systems. For this purpose, stapedius muscle contractions were elicited by high-level contralateral sound, controlled changes in middle ear pressure (range +/-100 daPa) were produced and the tympanic membrane was loaded with water droplets. A computer model of the middle ear network was implemented using a standard lumped-element electric analog of the middle ear (Zwislocki's model). Forward and backward transmission changes were simulated and model predictions were compared to experimental data. Apart from the case of positive middle ear pressures, a close qualitative correspondence was found between model and real-ear results. Each of the effects was characterized by a unique pattern of phase and magnitude changes as a function of frequency, in relation to the mechanical characteristics of the involved subsystem (i.e. stapes stiffness, tympanic-membrane stiffness or mass) and its resonance properties. Owing to their high sensitivity, EOAEs could be helpful for an accurate individual multifrequency analysis of middle ear impedance by comparisons under rest and test conditions. PMID- 10675647 TI - Middle-ear influence on otoacoustic emissions. II: contributions of posture and intracranial pressure. AB - Although it seems likely that body tilt or surgically provoked variations in intracranial pressure (ICP) can result in variations of intralabyrinthine pressure, the channels for pressure transmission remain controversial and the reasons why evoked otoacoustic emissions (EOAEs) exhibit attendant modifications are unclear. The theoretical framework implemented in the companion paper [Avan et al. part I, 2000] provides sensitive and non-invasive means to identify the middle-ear mechanism(s) entailed in EOAE changes. It was thus applied to analyze the influence of posture on EOAE phases and magnitudes as a function of frequency, in a series of experiments involving body tilt from sitting to supine (0 degrees or -30 degrees ). Controlled ICP variations were surgically carried out in a series of hydrocephalic patients and the resulting EOAE changes were compared to posture data and model predictions. In all cases, the EOAE changes closely resembled those due to an increase in the stiffness of the stapes' annular ligament, in keeping with the assumption that ICP gets transmitted to intralabyrinthine spaces and modifies the hydrostatic load on the stapes, thereby influencing EOAE features. A small additional contribution of middle-ear pressure to EOAE changes was identified in addition to the main stapes component. Dynamical EOAE measurements showed that sudden ICP changes were transmitted to the inner ear within 8-30 s. The high sensitivity of EOAE phases below 2 kHz to ICP changes, together with the absence of any significant confounding middle-ear effect, favors EOAEs for a reliable non-invasive monitoring of ICP and intralabyrinthine pressures. PMID- 10675648 TI - Technical report: chronic and acute intracochlear infusion in rodents. AB - As a follow-up to the Brown et al., 1993 technique, we have made several improvements to the micro-cannula, osmotic pump procedure, enabling chronic intracochlear infusions while maintaining hearing. In addition, acute bolus injection techniques are briefly described in guinea pig, rat and mouse. PMID- 10675649 TI - Amyloid precursor protein gene isoforms in Alzheimer's disease and other neurodegenerative disorders. AB - Differential expression of the amyloid precursor protein gene (APP) may be important in the development of amyloidosis in Alzheimer's disease (AD) and experimentally in the brain's response to injury. Controversial data suggests that APP isoforms containing the Kunitz protease inhibitor isoform (APP KPI+) are over expressed in the brains of patients with AD when compared to the non-Kunitz protease inhibitor containing isoforms (APP KPI-). We have investigated this hypothesis using a quantitative analysis of gene expression on brain tissue collected at post-mortem. In situ hybridization has been used with synthetic oligonucleotide probes labelled with 35S to detect the two principal splice variants of APP: APP 695 (KPI-) and APP 751 (KPI+). A prospective brain bank of frozen brain specimens has been established and includes pathologically proven AD (n=15) and other neurodegenerative disorders as controls (n=18). The controls consist of frontal lobe atrophy (n=4), Huntington's disease (n=5), Parkinson's disease (n=4), motor neuron disease (n=2), multi-infarct dementia (n=1), multisystem atrophy (n=1), and subacute sclerosing panencephalitis (n=1). We have observed no significant differences in the expression of APP 695 KPI- mRNA in frontal lobe: 17.49+/-3.26 optical density (OD) units of mRNA expression in AD vs. 16.13+/-1.76 OD units mRNA in controls (P=0.80, linear regression); or temporal lobe: 14.73+/-2.96 in AD vs. 16.49+/-2.15 in controls (P=0.55). No significant differences have been found in APP 751 KPI+ in frontal lobe: 12.86+/ 2.98 in AD vs. 13.70+/-2.88 in controls (P=0.97); and temporal lobe: 13.31+/-4.93 in AD vs. 11.07+/-1.99 in controls (P=0. 65). Analysis of the ratios of APP 751 KPI+ OD units of mRNA to APP 695 KPI- mRNA revealed a trend to an increased ratio which did not reach statistical significance: frontal lobe APP 751 KPI+/APP 695 KPI- 1.92+/-1.04 in AD vs. 0.86+/-0.17 in controls (P=0.54); temporal lobe 2.54+/ 1.59 in AD vs. 0.96+/-0.11 controls (P=0.34). Our data has not revealed differential expression of APP mRNA isoforms in AD and supports the hypothesis that post-translational events in APP metabolism are important in amyloidogenesis and the pathogenesis of AD. PMID- 10675650 TI - Inhibition of synaptosomal [3H]glutamate uptake and [3H]glutamate binding to plasma membranes from brain of young rats by glutaric acid in vitro. AB - Synaptosomes and plasma membrane preparations from brain of 30-day-old rats were incubated with glutaric acid at final concentrations ranging from 10 nM to 1 mM for the determination of glutamate uptake and binding, respectively. [3H]Glutamate uptake into synaptosomes was inhibited by approximately 50% by 1 mM glutaric acid, corresponding to the concentration found in brain of glutaric acidemic children. In addition, in the presence of extracellular Na+ concentrations, the same dose of glutaric acid decreased by about 30% [3H]glutamate binding to brain plasma membranes. The results indicate that the inhibition of both glutamate uptake into synaptosomes and glutamate binding to plasma synaptic membranes by the metabolite could result in elevated concentrations of the excitatory neurotransmitter in the synaptic cleft, potentially causing excitotoxicity to neural cells, a fact that may be related to the brain damage characteristic of glutaric acidemia type I. PMID- 10675651 TI - Detection of DNA fragmentation of myonuclei in myotonic dystrophy by double staining with anti-emerin antibody and by nick end-labeling. AB - To clarify the occurrence of apoptosis in skeletal muscle in pathological conditions, we studied 44 muscle biopsy specimens by immunohistochemical staining with monoclonal antibody against emerin, which is localized in muscle nuclear membrane, and by ApopTag Plus to detect DNA fragmentation. Five of six patients with myotonic dystrophy (DM) showed three to 35 myonuclei stained with anti emerin antibody and ApopTag Plus in 1500 muscle fibers. Four of the 18 patients with polymyositis, one of those with thyroid myopathy and one with neurogenic atrophy showed a few myonuclei stained positively by these methods. Our study revealed that DNA fragmentation in myonuclei occurred in skeletal muscle fibers regardless of the type of disease, although the frequency was rather low in all of these diseases except DM. The DNA fragmentation detected in most of the patients with DM suggested a significant role of apoptosis in the pathomechanism of this disease. PMID- 10675652 TI - Lesions visualized by contrast-enhanced magnetic resonance imaging in transient ischemic attacks. AB - In patients with transient ischemic attacks (TIAs), contrast-enhanced magnetic resonance imaging (MRI) is more sensitive to visualize the recent ischemic lesions than conventional MRI. We examined the clinical characteristics of TIA patients presenting with enhanced lesions visualized by contrast-enhanced MRI. We retrospectively evaluated 64 patients with carotid TIAs. We evaluated the frequency and topography of TIA associated infarcts on contrast-enhanced MRI and compared the clinical background of patients with and without such lesions. Twenty-three patients underwent plain MRI only, while the remaining 41 patients underwent contrast-enhanced MRI. Of the latter 41 patients, 16 had abnormal enhanced lesions (39%: group L), while 25 had no lesions (61%: group NL). In group L, all lesions were spotty, and they were located in the cerebral cortex in 13 patients (81%), the subcortex in two (12%), and the perforator territory in one (6%). Aphasia or confusional state, hypertension, and emboligenic cardiac or arterial disease (stenosis > or =50%) were more frequently observed in group L than in group NL (38 vs. 8%, 81 vs. 48%, and 93 vs. 60%, respectively, P<0. 05). The TIA patients with enhanced lesions on MRI may be associated with an emboligenic cardiac or arterial disease, severe neurologic symptom compared to those without them. PMID- 10675653 TI - Cerebral atrophy in multiple system atrophy by MRI. AB - Cranial magnetic resonance images (MRI) of the cerebral areas of 40 patients with multiple system atrophy (MSA) and of 61 age-matched controls were analyzed. The cerebral area of MSA patients was 131. 95+/-15.89 cm(2) (mean+/-S.D.), which was significantly smaller than that of normal controls at 149.01+/-10.93 cm(2) (P<0.0001). All 23 MSA cases subjected to the MRI study over a 1-year period showed progressive cerebral atrophy, and the atrophy rate was 2.46+/-1. 66%/year. There were no significant differences within the MSA subtypes or between gender. The progression of cerebral atrophy in MSA correlated more with duration (r= 0.634) than age (r=-0.421). We conclude that MRI findings throughout the course of MSA suggest progressive cerebral atrophy, which is common in all subtypes and reflects duration of the disease rather than age. PMID- 10675654 TI - The recovery potential of central conduction disorder in hypothyroid rats. AB - In an aim to detect the dysfunction of central nervous system among rats with varied durations of hypothyroidism and to elucidate the recovery potential after thyroxine replacement, a series of BAEP were conducted and compared with age matched controls. BAEP was performed in five groups of the hypothyroid animals 1, 3, 5, 7, and 9 months after thyroidectomy respectively. Following initial electrophysiological assessment, thyroxine replacement was administered to each group of hypothyroid rats, and BAEP was performed at two month intervals, up to two successive normal studies or six months after the initiation of therapy, whichever came first. Before thyroxine treatment, prolonged I-V interpeak latency was the most consistent abnormal finding in all groups of hypothyroid rats, and longer hypothyroid state correlated well with more severe central conduction disorder. Hearing impairment was also noted among those with long duration of hypothyroidism. After thyroxine replacement, the central conduction dysfunction usually returned to normal if the hypothyroid state was not more than 5 months in duration. However, when hypothyroid state persisted over 7 months or more, there would be an incomplete recovery for central conduction disorder. The present study brings out the concept of 'therapeutic window' in reversing the central nervous dysfunction caused by hypothyroidism in adult rats. PMID- 10675655 TI - Vergence disorders in patients with spinocerebellar ataxia 3/Machado-Joseph disease: a synoptophore study. AB - Diplopia, a common symptom in spinocerebellar ataxia 3/Machado-Joseph disease (SCA3/MJD) cases, is not always due to asymmetric ophthalmoplegia. We found a Japanese SCA3/MJD family, in which three patients clearly had an impairment of divergence eye movement. We thus quantitatively examined the vergence ranges in eight Japanese SCA3/MJD cases using the synoptophore test. An impairment of the vergence eye movements was found in all patients, and the vergence impairment pattern, but not the ophthalmoplegia pattern, was found to be compatible with the diplopia pattern. The diplopia in SCA3/MJD cases is, therefore, attributed, at least in part, to the impairment of the vergence eye movements. PMID- 10675656 TI - What's in a smile?: Quantification of the vertical smile of patients with myasthenia gravis. AB - Many patients with myasthenia gravis who experience bulbar symptoms show a vertical smile, which may have a considerable, and often underestimated, impact on social life. Peri-oral muscle function can be quantified by calculating lip length and snout indices, which indicate the degree to which a person is capable of smiling and of pursing the lips, respectively. In the present study patients with bulbar myasthenia gravis were compared to patients with ocular myasthenia gravis, patients now in remission (but previously suffering from bulbar myasthenia gravis), and healthy subjects. The lip-length and snout indices of patients with bulbar myasthenia gravis were significantly lower than those of the other groups. The facial impairments were no longer detectable in patients with bulbar myasthenia gravis in remission and no subclinical impairments in lip length and snout indices were found in the ocular myasthenia gravis group. These findings were consistent with the patients' reports of impairment of smiling and other oral functions. The patients suffering from a vertical smile or other oral impairments were well aware of their condition, most probably because of the social consequences of being unable to smile. The indices could be of importance in the longitudinal evaluation of therapy in individual patients and in pharmacotherapeutical research. We found a low correlation between the lip-length and snout indices, which reflects the capricious pattern of involvement of separate muscles in myasthenia gravis. Therefore both indices deserve special attention if they are used for monitoring myasthenic symptoms. PMID- 10675657 TI - The spectrum of chronic inflammatory demyelinating polyneuropathy. AB - Research criteria for the diagnosis of chronic inflammatory demyelinating polyneuropathy (CIDP) were proposed by an Ad Hoc Subcommittee of the American Academy of Neurology (AAN) in 1991, and since then these criteria have been widely used in clinical studies. We have been impressed by the frequent finding of electrophysiological changes of a demyelinating neuropathy in patients whose clinical presentation does not conform to the usually accepted clinical phenotype of CIDP. To determine the clinical spectrum of CIDP, we conducted a retrospective review of patients of the peripheral electrophysiology laboratory of the University of Miami-Jackson Memorial Medical Center. Diagnostic criteria for acquired demyelination of an individual nerve were adapted from the AAN research criteria for the diagnosis of CIDP (1991). Patients were accepted for inclusion when such evidence was demonstrated in at least one motor nerve or at least two sensory nerves. We then reviewed the clinical phenotype and the underlying etiology of the neuropathy in these cases. Eighty-seven patients, 63 male and 24 female, age of onset 4-84 (mean 49.3) years, met these inclusion criteria. Forty seven patients (54%) had distinct features outside the usual clinical presentation of CIDP. Of these, 15 (17%) had predominantly distal features, 13 (15%) had exclusively sensory polyneuropathy; seven (8%) had markedly asymmetric disease, seven (8%) had associated CNS demyelination, four (5%) had predominant cranial nerve involvement, and one (1%) had only the restless legs syndrome. An associated medical condition that may have been responsible for the acquired demyelinating neuropathy was present in 60% of the patients. We conclude that spectrum of CIDP is broader than would be indicated by the strict application of the AAN research criteria, and that many of the cases meeting more liberal criteria frequently respond to immunosuppressive therapy. PMID- 10675658 TI - Migraine, but not subarachnoid hemorrhage, is associated with differentially increased NPY-like immunoreactivity in the CSF. AB - To test whether migraine and subarachnoid hemorrhage (SAH) are associated with increased sympathetic tone, we compared the neuropeptide Y-like (NPY-LI) and chromogranin A-like immunoreactivities (LI) of cerebrospinal fluid (CSF) from migraneurs and SAH patients with those from control subjects. Increased sympathetic tone was expected to produce higher co-release of these co-stored peptides and concordant changes in their CSF levels. In addition, we investigated a possible disturbed nitric oxide homeostasis by measuring CSF nitrites (NO). More than 70% of CSF NPY-LI corresponded to the chromatographic peak (HPLC) for the intact molecule in all three groups. Migraneurs had 64% higher CSF NPY-LI, but no significant difference in CSF chromogranin A-LI, as compared to controls. In contrast, SAH patients had 74% less CSF chromogranin A-LI and a trend to lower NPY-LI, as compared to controls. No differences in CSF NO were detected among groups. These results argue against an increased sympathetic tone in patients with either migraine or SAH, and suggest that the higher CSF NPY-LI of migraneurs probably originates from central neurons. Furthermore, our findings in SAH patients argue in favor of a decreased sympathetic tone; this could be a homeostatic response to counterbalance vasoconstriction mediated by other mechanisms. PMID- 10675659 TI - Analysis of the mRNA transcripts of the survival motor neuron (SMN) gene in the tissue of an SMA fetus and the peripheral blood mononuclear cells of normals, carriers and SMA patients. AB - Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of the anterior horn cells of the spinal cord. The gene most highly associated with SMA is the survival motor neuron (SMN) gene. In this study, we present an analysis of messenger RNA (mRNA) expression of the SMN gene in peripheral blood mononuclear cells in normal subjects, SMA carriers and patients from 20 SMA families. We found at least 6-8 different transcripts of SMN gene formed by alternative splicing involving exons 3, 5 and 7. We compared transcripts from the different types of SMA and found no definite differences in transcript patterns and amounts. Normal subjects with the telomeric SMN (SMN(T)) gene only had variable splicing resulting in several transcripts, the most dominant being a transcript containing all coding regions. However, SMA patients with the centromeric SMN (SMN(C)) gene only had a higher degree of splice variation and tended to show little or no exon 7. These results demonstrate that SMN(T) and SMN(C) genes participate in alternative splicing phenomena. The different splicing patterns support the view that the SMN(T) gene is responsible for SMA disease. We also analyzed the transcripts from several tissues of an SMA fetus who had a homozygous SMN(T) gene deletion. Different splicing patterns were also found in these tissues, and were similar to the splicing pattern of leukocytes. We compared the major transcripts from exons 4 to 8 of both the SMN(T) and SMN(C) genes and found that the relative proportion varied among normal subjects, SMA carriers and patients. This approach could be used as a novel diagnostic method. We suggest that analyzing the mRNA expression of the SMN gene in peripheral blood mononuclear cells offers an apparently reliable technique for separating SMA patients, carriers, and normal individuals. PMID- 10675660 TI - A new diagnostic procedure to detect unknown transthyretin (TTR) mutations in familial amyloidotic polyneuropathy (FAP). AB - Two patients with amyloidosis caused by transthyretin (TTR) were investigated by immunohistopathologic, mass spectrometric, and molecular genetic methods. After confirming the immunoreactivity of TTR in the amyloid deposits using anti-TTR polyclonal antibody, a new method: centrifugal concentration and electrospray ionization mass spectrometry (ESI-MS) was employed to detect the variant TTR in the serum. Only 50 microl of the serum and 30 microl of the anti-TTR antibody were needed for the analysis. After incubation with the antibody, the samples were passed through a 1000 kDa cut off centrifugal concentrator to retain the antibody, thereafter, the filtrate was analyzed by ESI-MS. Several forms of normal and variant TTR were detected in the serum samples: unconjugated TTR, cysteine and cysteine-glycine conjugated TTR. In the patients, a variant form of TTR was detected with a 26.0 Da higher molecular weight than that of normal TTR. Single-strand conformation polymorphism (SSCP) and direct sequence analysis confirmed the presence of a one-base substitution situated at the codon 50 from AGT (Ser) to ATT (Ile) in both patients, that corresponded to the increased molecular weight of 26.0. The present diagnostic procedure demonstrates the usefulness of both ESI-MS and SSCP to screen for TTR related amyloidosis rapidly. Moreover, the DNA samples obtained from the band showing abnormal electrophoretic migration pattern in SSCP, facilitate the direct sequence analysis to detect the unknown mutation, and the observed shift in molecular weight of the variant TTR in ESI-MS confirms the base substitution. PMID- 10675661 TI - Factor structure of trunk performance data for healthy subjects. AB - OBJECTIVE: To examine the factor structure of various measurements of trunk muscle performance for healthy subjects. DESIGN: A total of 22 performance scores were collected and their univariate and multi-variate relationships were examined. BACKGROUND: Extensive literature exists on the measurement of trunk performance data and the relationships between measurements but what needs to be collected to realize a true performance score remains unclear. METHODS: Trunk muscle performance scores of 150 subjects (71 males and 79 females) were obtained on an Isostation B-200 Dynamometer. Twenty-two parameters measuring range of motion, isometric strength, velocity, and endurance on all three planes of motion were collected. The factor structures were constructed using Principal Components Analysis. RESULTS: Clear-cut factor patterns (explained 96.3% of the total variance) suggests that the five-factor structure might be valid and appropriate for this population. The major loading on each factor indicated that: Factor 1 could be labeled as a static strength measure; Factor 2 as velocity; Factor 3 as flexibility; and Factors 4 and 5 as fatigue-resistance. CONCLUSIONS: No single mode of measurement can provide a good representation of a total trunk muscle performance. RelevanceFor the realization of trunk muscle performance, clinics have to measure all modes of isometric strength, velocity, range of motion, and endurance. Care must be taken in eliminating any parameter. PMID- 10675662 TI - Ultrasonic quantification of osseous displacements resulting from skin surface indentation loading of bovine para-spinal tissue. AB - OBJECTIVE: To validate an ultrasound-based technique which quantifies uni-planar subcutaneous displacement of an osseous object resulting from an externally applied load. BACKGROUND: Many spinal conditions are thought to be characterized by aberrant vertebral displacements yet the invasive nature of many investigative techniques has left the clinical significance of this relation incompletely understood. METHOD: Six bovine bone/paravertebral tissue preparations were indented by one of two ultrasonic transducers (5 and 7 MHz) fitted to an electromechanical actuator. The resulting osseous displacement along the principal indentation axis was calculated by subtracting the change in transducer/bone distance between ultrasonic images collected at tissue contact and maximal load from the change in actuator displacement. A dial gauge contacting the bone was used as a displacement criterion measure. RESULTS: Using the 7 MHz transducer, the mean error of the technique was 6.74% (SD=3.98) while the mean error associated with the 5 MHz transducer was 12.73% (SD=7.49). CONCLUSIONS: This non-invasive technique is capable of quantifying subcutaneous uni-planar bone displacement with an accuracy comparable to similar invasive techniques over a comparable displacement range. RelevanceThis non-invasive technique may be beneficial in assessing the significance of vertebral displacements in conditions such as hypermobility and osteoarthritis, as well as in studies of manipulative therapy. PMID- 10675663 TI - Description of the relation between the forces acting in the lumbar spine and whole-body vibrations by means of transfer functions. AB - OBJECTIVE: The purpose of this study was to display the relationships between the forces transmitted in the spine and the accelerations of the vibrating seat. BACKGROUND: Investigations reveal that exposure to whole-body vibration can induce degenerative changes in the lumbar spine. Elevated spinal forces are probably the crucial component in the pathogenesis of this disease. DESIGN AND METHODS: The spinal forces are simulated by means of a biomechanical model, where 16 rigid bodies represent the upper body and the arms of a sitting operator. The relationships between seat accelerations and spinal forces are displayed as frequency-dependent transfer functions. RESULTS: Spinal forces are not only elevated in the direction parallel to the vibration excitation but also in the two other orthogonal vibration directions. According to the magnitudes of the transfer functions the highest oscillating parts of the forces are reached at frequencies below 10 Hz. CONCLUSIONS: Using the transfer functions, the time course of the spine forces can be computed and a new kind of weighting function can be derived which enables a force related weighting of the seat acceleration. RelevanceVibration induced health risks are commonly assessed by the weighted acceleration (ISO 2631-1). These weighting factors resulted from subjective magnitude sensation. It is argued that a more valid assessment will be obtained if the accelerations are weighted in relation to spinal forces. These forces cannot be measured directly under vibration. However, they can be simulated by means of biomechanical models. PMID- 10675664 TI - The bursal and articular sides of the supraspinatus tendon have a different compressive stiffness. AB - OBJECTIVE: To measure the compressive stiffness of the supraspinatus tendon and to determine whether regional difference exists in the bursal and articular side of the tendon. DESIGN: Indentation testing was performed on both the bursal and articular sides of the supraspinatus tendon, focused on the 'critical area', where rotator cuff tears often occur. BACKGROUND: When the supraspinatus tendon wraps around the humeral head or is under impingement condition, compressive force on the tendon surface is expected. Therefore, compressive stress has been recently considered to be one of the important factors associated with the cuff tear. The mechanical properties would be essential for analytic modeling of stress distribution. METHODS: Indentation tests were performed at 15 locations on the bursal and articular surfaces of the supraspinatus tendon. A mathematical model with exponential relationship was used to describe the measured force deformation relationship and to calculate the compressive stiffness of the supraspinatus tendon. RESULTS: The over-all initial stiffness on the bursal and articular sides of the tendon was significantly different. On the bursal side, the anterior third had a significantly higher initial stiffness than the other thirds on average. On the articular side, initial stiffness at location 10 mm proximal to the greater tuberosity was significantly higher than the rest on average. CONCLUSIONS: The compressive stiffness of the supraspinatus tendon was found to be non-homogenous throughout the structure.RelevanceNon-homogenous compressive stiffness of the supraspinatus tendon would affect the load transmission within the tendon, which might be associated with the potential mechanism of tear. Such characteristics needs to be considered when performing finite element modeling of stress fields in the tendon. PMID- 10675665 TI - Three-dimensional measurement of cemented femoral stem stability: an in vitro cadaver study. AB - OBJECTIVE: To compare the in vitro stability of two cemented hip stem designs: Stem I was a collarless, double-tapered, highly polished implant; Stem II had a collar and matt finish. BACKGROUND: Stability of the femoral component of a hip implant is important for its long-term clinical success. Excessive migration or cyclic motion can increase the risk of early implant failure. METHODS: The stems were implanted in paired human cadaver femurs, and custom-designed micromotion sensors were used to measure three-dimensional motions of the stems at proximal, middle and distal locations during simulated in vivo loading cycles. RESULTS: This study found that despite 'rigid' fixation, cemented stems exhibit detectable motions under a limited number of cycles of simulated physiologic loads. At four times the donor body weight, Stem I showed a subsidence of 90 microm, compared to 25 microm of Stem II (P<0.05). In contrast, the proximal end of Stem II exhibited greater cyclic motions in the medial-lateral direction (P<0.05). CONCLUSIONS: The different motion patterns could be due to the design differences, such as surface finish and geometry. RelevanceImplant design is an important factor related to the behavior of the cement/bone interface and the overall success of the implant. This study compares in vitro micromotion of two cemented femoral prostheses with differing proximal designs. PMID- 10675666 TI - Determination of hydrodynamic drag forces and drag coefficients on human leg/foot model during knee exercise. AB - OBJECTIVE: The purpose of this laboratory experiment was to measure hydrodynamic drag forces in barefoot/hydro-boot conditions and accordingly, to determine the coefficients of drag on human leg/foot model during simulated knee extension flexion exercise. DESIGN: The prosthesis of the human lower leg was set in a water tank and connected into an isokinetic force dynamometer to measure resistive forces during knee motion. BACKGROUND: Quantifying resistance for aquatic exercises has been a challenge in hydrotherapy. The use of models of foot/leg provides a practical method to calculate coefficients of drag and to estimate resistance for rehabilitation purposes in musculoskeletal and amputee patients. METHODS: The dynamometer produced constant angular velocities of 250 degrees /s, 270 degrees /s and 300 degrees /s to the prosthesis. The baseline for measurements was performed in barefoot condition. A hydro-boot was used to study effects of increased frontal area (30%) of the leg on drag forces and coefficients. RESULTS: The maximal drag force values were 61 N (300 degrees /s) in barefoot and 270 N (270 degrees /s) in hydro-boot condition. Related drag coefficient values during the range of motion were from 0.3 to 0.1 and from 1 to 0.8, respectively. CONCLUSIONS: Drag force and related drag coefficient were highest during the early part of extension (150-140 degrees flexion) as the model was opposing the lift forces with the influence of water resistance. The effect of velocity was remarkable on drag forces but minimal on drag coefficient values. RelevanceThe drag forces and coefficients of this experiment can be clinically utilised to calculate hydrodynamic forces to develop progressive knee exercise programs as well as to design of prosthesis for amputee patients. PMID- 10675667 TI - Passive stiffness characteristics of ankle plantar flexors in hemiplegia. AB - OBJECTIVE: To assess the feasibility and reliability of ankle plantar flexor stiffness measurements in hemiplegia. DESIGN: Repeated measurements in five consecutive weeks. BACKGROUND: In hemiplegia, an equinovarus positioning of the foot might be caused by an increased stiffness of the m. triceps surae. METHODS: In eight hemiplegic patients the net joint torque of passive muscle stretch was measured as a function of ankle-angle by a dynamometer, at both sides. Ankle stiffness was characterised and also a biomechanical model of the passive muscle was fitted. RESULTS: In the vast majority of measurements it was possible to obtain measurements that were not distorted by involuntary muscle contraction. These measurements showed for the angle at which a passive plantar moment of 10 N m was reached a standard error of measurement of less than 2.7 degrees. The muscle model showed the increased stiffness as a shortening of the muscle-fibre length. CONCLUSION: The feasibility of this method to measure muscle stiffness was fair to good in hemiplegic patients. Provided the abandoning of involuntary muscle activity, the reproducibility warrants application in clinical practice at an individual level. The use of the model relates this changes to a shortened m. soleus and/or m. gastrocnemius. RelevanceEffective clinical decisions for treatment of equinovarus positioning of the foot in the hemiplegic individual, should consider excessive involuntary contractions of the m. triceps surae complex (i.e., spasticity), as well as shortened muscle tissue resulting in high stiffness. Despite the importance of increased stiffness there have been no validated methods of measurement. PMID- 10675668 TI - Foot progression angle and ankle joint complex in preschool children. AB - OBJECTIVE: The influence of foot progression angle on the ankle joint and the effects on gait patterns and mechanisms in skeletally normal preschool children was investigated. DESIGN: Kinematics and kinetics of the ankle joint were analyzed for preschool children who were skeletally normal but walked with different foot progression angle. BACKGROUND: The most frequent reasons for preschool children to be brought to a paediatric orthopaedic clinic are toe-in and toe-out. Without understanding the biological and biomechanical implications, treatment for these problems can be very confusing. METHODS: Gait analysis was performed in 86 skeletally normal preschool children. Children were grouped according to their foot progression angles. Analysis of the kinematics and kinetics of the ankle joint was intended to elucidate the gait mechanism. RESULTS: Children with different foot progression angles had distinctive patterns of spatio-temporal parameters, ground reaction force, joint angle, moment, power, and mechanical work of the ankle joints. The differences were organized and explained as different walking patterns and tactics. CONCLUSIONS: Skeletally normal preschool children with excessive toe-in or toe-out foot progression angles are not necessarily in some diseased status. They were instead related to different walking patterns. Aggressive treatment for these problems is not recommended.RelevanceThese results suggest that toe-in and toe-out are related to walking speed which has distinct influences on the kinematics and kinetics of the ankle joint. Though the observed problems were seemingly only in the transverse plane, they are in fact three-dimensional and have a mutually close relationship. The findings could be linked to the promptness of muscle response and the habits of walking in preschool children. Better understanding of possible mechanisms will help parents and paediatricians decide on the most appropriate treatment for these children. PMID- 10675669 TI - A study of in-shoe plantar shear in patients with diabetic neuropathy. AB - OBJECTIVE: To quantify in-shoe plantar shear in diabetic neuropathic feet. DESIGN: Plantar shear stresses are measured in a group of six patients with a history of diabetic neuropathic ulceration. BACKGROUND: Although elevated pressure between foot and shoe frequently found in diabetic neuropathic patients has been linked to a raised incidence of plantar ulceration, the shear component of stress at this interface is as yet unquantified. It is suggested that its effects may be equally damaging. METHODS: Measurements of shear were made locally beneath the medial four metatarsal heads and heel during unpaced gait in orthopaedic footwear, using a bi-axial magneto-resistive shear transducer. Similar methodology was previously employed on a group of asymptomatic adults, thereby allowing comparisons to be made. RESULTS: Overall the maximum shear stress for this patient group (73 kPa) was not significantly different to that in the asymptomatic group (87 kPa). However the patient group exhibited lower magnitudes of shear stress under the third/fourth metatarsal heads (average 51/39 vs. 86.5/71 kPa, respectively) and higher magnitudes under the first/second heads (73/64 vs. 35/31 kPa, respectively), indicating a medial shift. Step-to-step variability of maximum shear measured under the third metatarsal head showed an increase in the transverse component (coefficient of reliability 67% vs. 98%). CONCLUSIONS: Although the overall patterns of shear are broadly similar to the asymptomatic group, these pilot trials indicate a medial shift in shear loading under the forefoot coupled to increased step-to-step variability in the diabetic group. RelevanceMechanical stress at the plantar interface between foot and shoe is of particular clinical relevance to the formation and management of ulcers in diabetic neuropathy. Whereas the pressure component of stress is widely studied, the shear component is poorly described although it may be of equal importance. PMID- 10675670 TI - Mechanical properties of collagen fascicles from in situ frozen and stress shielded rabbit patellar tendons. AB - OBJECTIVE: To know the effects of stress shielding on the biomechanical properties of collagen fascicles obtained from in situ frozen patellar tendons (an autograft model). DESIGN: Collagen fascicles of approximately 300 microm in diameter were obtained from in situ frozen rabbit patellar tendons and also from in situ frozen and stress-shielded ones, and their mechanical properties and fibroblast density were determined. BACKGROUND: Stress shielding changes the mechanical properties of in situ frozen patellar tendons in which there exist no fibroblasts. The mechanisms of this phenomenon have not been studied well. METHOD: Patellar tendons of both in situ frozen group and in situ frozen and stress-shielded group were frozen in situ by liquid nitrogen to kill fibroblasts. Then, in the in situ frozen and stress-shielded group, no tension was applied to the tendons for 2, 3, and 6 weeks, while normal tension was applied to the tendons of the in situ frozen group. Tensile properties of the collagen fascicles obtained from these tendons were determined using a microtensile tester, and were compared to the collagen fascicles from non-frozen, stress-shielded patellar tendons. RESULTS: Tangent modulus and tensile strength of collagen fascicles from the in situ frozen and stress-shielded group progressively decreased with the time of stress shielding; however, these decreases were much smaller than those of the fascicles obtained from non-frozen, stress-shielded tendons. Although there were few fibroblasts in the patellar tendon of the in situ frozen and stress-shielded group at 2 weeks, the modulus and strength of the fascicles from the posterior portion were significantly lower than those in the in situ frozen group. In addition, the reduction of strength caused by stress shielding was much smaller in collagen fascicles than in bulk patellar tendons. CONCLUSION: The mechanical properties of collagen fascicles in in situ frozen tendons (an autograft model) are affected by stress shielding even under acellular condition. RelevanceThe in situ frozen, stress-shielded patellar tendon is a model of augmented autografts which are clinically used for the reconstruction of injured anterior cruciate ligaments. The sub-macroscopic studies of the tendon are useful to understand the mechanisms of the reduction of graft strength and its gradual recovery observed after reconstruction. PMID- 10675671 TI - The effect of lumbar back support tension on trunk muscle activity. AB - OBJECTIVE: Assess the effect of different controlled lumbar back support tightness levels on trunk muscle activity. DESIGN: Two-way repeated measure design assessing lumbar back support tension and submaximal trunk extension moments on trunk muscle electromyographic activity. BACKGROUND: Biomechanical studies on lumbar back supports often use electromyography (EMG) to assess the affect on trunk muscle activity. However, the lumbar back support may alter the electromyographic signal by changing the electrode-muscle distance. METHODS: Subjects performed trunk extensions at three static submaximal extension moment levels (25%, 50% and 75% MVC) while stabilized at the hips and shoulders, with the back support tensioned to three different tightness levels (44.5, 66.7 and 89.0 N) as well as a no-back support condition. RESULTS: Statistical analysis failed to find a significant effect (P8 h), the drug content was also increased. The drug release from those tablets was sufficiently sustained (more than 8 h) and non-Fickian transport of the drug from tablets was confirmed. Radiological evidence suggests that, that the formulated tablets did not adhere to the stomach mucus and that the mean gastric residence time was prolonged (>4 h). PMID- 10675691 TI - An investigation into the release of cefuroxime axetil from taste-masked stearic acid microspheres. II. The effects of buffer composition on drug release. AB - The influence of buffer composition on the release of cefuroxime axetil from stearic acid microspheres has been investigated, with particular emphasis on establishing the relationship between buffer composition and release at a single pH value. Studies of drug dissolution and release from spheres in pH 7.0 citrate phosphate buffer (CPB), boric acid buffer (BAB), phosphate buffer mixed (PBM) and Sorensens modified phosphate buffer (SMPB) indicated marked differences in release profile from the spheres, with an approximate rank order of SMPB > CPB approximately BAB > PBM. The role of added sodium was then investigated by examining the release profiles in SMPB and PBM to which sodium ions had been added. Increases in the sodium content from approximately 0.11 to 0.2 M were found to decrease the release rate for the SMPB, while increases from 0.007 to 1.0 M sodium in PBM resulted in a maximum release being seen for the systems containing 0.05 M sodium. Studies on surface disintegration, using scanning electron microscopy (SEM) and sodium uptake using flame emission spectroscopy, indicated an interrelationship between medium composition, disintegration and release. The data are discussed in terms of the possible mechanisms associated with drug release from these spheres. PMID- 10675692 TI - Anti-mucus polyclonal antibody production, purification and linkage to the surface of albumin microspheres. AB - This aim of this study was to develop a microparticulate based oral drug delivery system, which could prolong gut transit time by binding via specific interactions to the gut mucus layer. Porcine gastric mucus was semi-purified and used as an antigen to raise a polyclonal antiserum in rabbits. The immunoglobulin fraction of this serum was isolated, purified and tested for homogeneity and cross reactivity. High cross-reactivity was displayed when the antiserum was challenged against types of mucus other than that used as an antigen, but no significant cross-reactivity occurred when challenged against some other common macromolecules. The antibody fraction of this serum was covalently linked to three types of albumin microspheres (MS) using 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide. The MS employed had either a hydrophobic, a hydrophilic or a carboxymethylated surface, and were prepared and characterised as described earlier (MacAdam, A.B., Shafi, Z.B., Martin, G.P. and James, S.L. 1997. Preparation of hydrophobic and hydrophilic MS and determination of surface carboxylic acid and amino residues. Int. J. Pharm. 151, 47-55). Binding of these MS to both radioiodinated mucin in suspension and to isolated gut segments was measured. Hydrophilic and carboxymethylated MS with surface-associated antibody bound significantly more mucin from a suspension than did uncoated controls. Similarly, anti-mucus antibody-coated hydrophilic and carboxymethylated MS bound more strongly to an isolated gut segment than did uncoated controls or controls coated in an antibody specific for albumin. These results suggest anti-mucus antibody coated albumin MS may be a useful model to act as comparators in studies aimed at developing drug delivery systems with delayed gastrointestinal transit. PMID- 10675693 TI - Artefact formation in the determination of residual solvents according to a method of the European Pharmacopeia. AB - Method 2 of the procedure for the identification and assay of residual solvents, of the European Pharmacopeia 3rd edition 1999 addendum, leads to artefactual formation of N-chlorodimethylamine when the hydrochlorides of basic compounds are examined. This is due to degradation of the dissolution solvent N,N dimethylformamide under the prescribed conditions. N-Chlorodimethylamine has been detected during analysis of several hydrochloride salts of nitrogen bases including drug substances. Artefact formation did not occur consistently with all the compounds examined, but with diltiazem hydrochloride it was observed in the majority of experiments. The discovery that the alkylating reagent N,N dimethylaminoethyl chloride (DMC) used in the synthesis of diltiazem gives apparently high yields of N-chlorodimethylamine was cause for concern. However, it has been confirmed that production batches of diltiazem hydrochloride contain <1 ppm of this synthetic intermediate. The formation of N-chlorodimethylamine in the presence of the drug substance is probably due to a reaction between dimethylformamide and HCl, that would be released as a result of hydrolysis by residual water of the O-acetyl function of diltiazem. In view of these findings, the compendial general method should be reviewed. It may be necessary to adopt a different approach to the drafting of methods for volatile impurities, since most of the operating conditions are in practice specific to the substance being examined. PMID- 10675694 TI - In vitro permeation through porcine buccal mucosa of Salvia desoleana Atzei & Picci essential oil from topical formulations. AB - In the light of recent studies, which have shown that the essential oil derived from some Lamiaceae species has appreciable anti-inflammatory activity, moderate anti-microbial action and the ability to inhibit induced hyperalgesia, an assessment of the diffusion and permeation of Salvia desoleana Atzei & Picci (S. desoleana) essential oil through porcine buccal mucosa was considered useful for a possible application in the stomatological field. Topical formulations (microemulsions, hydrogels and microemulsion-hydrogels) were prepared for application to the buccal mucosa. The mucosa permeation of the oil from the formulations was evaluated using Franz cells, with porcine buccal mucosa as septum between the formulations (donor compartment) and the receptor phase chambers. The study also aimed at optimising the permeability of the S. desoleana essential oil by means of an enhancer, the diethylene glycol monoethyl ether Transcutol. The diffusion of the oil through the membrane was determined by evaluating the amount of essential oil components present in the receiving solution, the flux and the permeation coefficient (at the steady state) in the different formulations at set intervals. Qualitative and quantitative determinations were done by gas chromatographic analysis. All the formulations allow a high permeability coefficient in comparison with the pure essential oil. In particular, the components with a terpenic structure (beta-pinene, cineole, alpha-terpineol and linalool) have the highest capacity to pass through the porcine buccal mucosa when compared to the other components (linalyl acetate and alpha-terpinil acetate). Moreover, the enhancer, diethylene glycol monoethyl ether largely increases the permeation of the essential oil components in relation to the concentration. PMID- 10675695 TI - Diclofenac sodium incorporated PLGA (50:50) microspheres: formulation considerations and in vitro/in vivo evaluation. AB - Recently, considerable interest has been focused on the use of biodegradable polymers for specialized applications such as controlled release of drug formulations; meanwhile, microsphere drug-delivery systems using various kinds of biodegradable polymers have been studied extensively during the past two decades. Poly (lactide-co-glycolide) (PLGA) polymers have been proven to be excellent drug carriers for microparticulate systems due to their advantages, e.g. biocompatibility and regulatory approval. The administration of nonsteroidal anti inflammatory drugs (NSAIDs) into the intra-articular cavity in patients with chronic inflammatory disease is complicated due to the short duration of effect. In the present study, controlled-release parenteral formulations of diclofenac sodium (DS), a commonly used NSAID, were prepared for intra-articular administration, and evaluated in vitro for particle size, yield, drug loading, surface morphology and release characteristics. For in vivo studies, Technetium 99m labelled polyclonal human immunogammaglobulin (99m Tc-HIG) was used as the radiopharmaceutical to demonstrate arthritic lesions by gamma scintigraphy. Evaluation of arthritic lesions post-therapy in rabbits showed no significant difference in the group treated with PLGA (50:50) (mw 34000) DS microspheres compared to control groups. PMID- 10675696 TI - Unexpected skin barrier influence from nonionic emulsifiers. AB - Skin disorders are often treated with creams containing various active substances. The creams also contain emulsifiers, which are surface-active ingredients used to stabilize the emulsion. Emulsifiers are potential irritants and in the present study the influence of stearic acid, glyceryl stearate, PEG-2, -9, -40, and -100 stearate, steareth-2, -10 and -21 on normal as well as on irritated skin have been evaluated with non-invasive measurements. Test emulsions were created by incorporating 5% emulsifiers in a water/mineral oil mixture (50:50). The emulsions and their vehicle were then applied to normal skin for 48 h and to sodium lauryl sulfate (SLS) damaged skin for 17 h in aluminum chambers. Twenty-four hours after removal of the chambers the test sites were evaluated for degree of irritation. In normal skin, the emulsifiers induced significant differences in TEWL but not in skin blood flow. Five of the emulsifiers increased TEWL. In SLS-damaged skin an aggravation of the irritation was expected. However, no differences regarding skin blood flow was noted from the emulsifiers. Furthermore, three emulsifiers unexpectedly decreased TEWL. These results highlight the possibility of absorption of these emulsifiers into the lipid bilayer, which increase TEWL in normal skin and decrease TEWL in damaged skin. PMID- 10675697 TI - Nasal absorption of (S)-UH-301 and its transport into the cerebrospinal fluid of rats. AB - Targeting the brain via nasal administration of drugs has been studied frequently over the last few years. In this study, the serotonin-1a receptor antagonist (S) 5-fluoro-8-hydroxy-2-(dipropyl-amino) tetralin ((S)-UH-301) hydrochloride was used as a model substance. The systemic absorption and transport of (S)-UH-301 into male Sprague-Dawley rat cerebrospinal fluid (CSF) were investigated after nasal and intravenous administration. Blood and CSF samples were obtained at regular time intervals from the arteria carotis and by cisternal puncture, respectively, after administration to both nostrils (total 12 micromol/kg) or into the vena jugularis (6 micromol/kg). The concentrations of (S)-UH-301 in plasma and CSF were measured by HPLC with electrochemical detection. The maximum plasma concentration of intranasal (S)-UH-301 occurred in about 7 min and the absolute bioavailability seemed to be complete (F=1.2+/-0.4). Initially, no increased concentrations of (S)-UH-301 were seen in CSF after nasal compared to intravenous administration i.e. it appeared that no direct transport of (S)-UH 301 from the nasal cavity, along the olfactory neurons and into the CSF occurred. However, a prolonged duration of the concentration was seen after nasal administration of (S)-UH-301 and after about 20 min the CSF(na):CSF(iv) concentration ratio (corrected for different dosage) exceeded 1. PMID- 10675698 TI - The interaction of human serum albumin and model membranes. AB - It is frequently observed that the interaction of human serum albumin (HSA) with different lipid membranes may affect molecular transport both in vivo and in vitro experiments. There was a lack of consensus however in the interpretation of results. Earlier studies on the serum albumin membrane association had different conclusions depending on the source of protein, the preparation and the composition of the membranes applied. In this work the change of heat capacity, a sensitive parameter of the interacting system, is compared for uni- and multilamellar liposomes (dimyristoyl-phosphatidylcoline/dimyristoyl phosphatidylglycerol) at 0, 1x10(-3), 8x10(-3), 1.2x10(-2) and 3.3x10(-2) HSA lipid ratios. The thermal properties of the sonicated and vortexed liposomes show remarkable differences. The presence of HSA in both types of liposomes also modified their thermal properties, providing clear evidence for protein-vesicle interaction, different in the uni- and multilamellar liposomes. In the case of unilamellar liposomes, two additional transitions were observed at lower temperature, independently of the HSA-lipid ratio, and the protein binding mode to smaller or larger sized liposomes was also distinguishable. The addition of HSA to the multilamellar liposomes resulted in an increase of the pretransition temperature only at the higher HSA-lipid ratio, but the main transition temperature was not affected. PMID- 10675699 TI - Silica xerogel carrier material for controlled release of toremifene citrate. AB - Sol-gel processed silica xerogel was used as a carrier material for toremifene citrate in order to develop an implantable controlled release formulation which could be localised to a desired site providing targeted and long-lasting disease control and resulting in a reduced amount of drug needed. Toremifene citrate, an anti-estrogenic compound, was incorporated into silica xerogel matrixes during polycondensation of organic silicate, tetraethyl ortho silicate (TEOS). The effects of drug amount, drying temperature and polyethylene glycol (PEG) on the release rate of toremifene citrate and degradation of the silica xerogel matrixes were investigated. Addition of PEG (M(w) 4600/10000) decreased the specific surface area of the matrix and lowered the release rate of the drug. Reducing the amount of drug in the matrix also decreased the release rate of toremifene citrate. However, drying temperature did not affect the release rate of silica or toremifene citrate. The release profiles of toremifene citrate were according to zero order kinetics, suggesting that drug release was controlled by erosion of the silica xerogel matrix. These results suggest that the toremifene citrate release rate can be controlled to some extent by adding (PEG) or by varying the amount of drug in the silica xerogel matrix. PMID- 10675700 TI - High molecular weight polyethylene oxides (PEOs) as an alternative to HPMC in controlled release dosage forms. AB - High molecular weight polyethylene oxides (PEOs) have recently been proposed as an alternative to hydroxypropylmethylcellulose (HPMC) in controlled release matrix tablets. In this study, we compared the performance of PEO and HPMC polymers when employed in the Geomatrix technology, a versatile, well-known method to achieve extended release of drugs at a constant rate. Four core formulations were prepared, containing a soluble drug (diltiazem) and, alternatively, PEO or HPMC of two different viscosity grades. These formulations have the same composition except for the polymer employed. Similarly, four barrier formulations were also prepared, which only differ in the kind of polymer employed. Three-layer Geomatrix systems were then prepared using these core and barrier formulations. The release profiles of the different three-layer systems obtained were compared, to verify if PEO could efficiently replace HPMC in this type of dosage form. The results show that slower release rates can be obtained from the plain matrices containing HPMC compared to PEO, moreover HPMC, used in the barrier formulations, is generally more efficient in controlling drug release rate in three-layer Geomatrix systems. PMID- 10675701 TI - Preparation and characterisation of a new insoluble polymorphic form of glibenclamide. AB - A crystalline form of glibenclamide, with higher melting point (218 degrees C) and lower solubility in simulated gastric and intestinal fluids, was arisen during an attempt to elucidate transitional phases by melting, cooling and reheating. The new form was obtained from the glassy state, by applying sublimation at 130-160 degrees C and was characterised by differential scanning calorimetry (DSC), infrared (IR) spectroscopy, scanning electron microscopy (SEM), hot-stage microscopy (HSM), X-ray powder diffraction (XRD) and solubility studies. Formation of the new crystal form is considered as reason of reduction in dissolution and bioavailability of tablets. PMID- 10675702 TI - Surface drug removal from ibuprofen-loaded PLA microspheres. AB - The preparation, characterisation and drug release behaviour of ibuprofen loaded poly(D,L-lactic acid) (PLA) microspheres are described. Depending on the gelatin concentration in the aqueous external solution (1, 0.5, 0.1% w/v), microspheres with three different sizes (2.2, 4.1, 7.5 micrometer) were obtained. The properties of microspheres washed with water (Untreated microspheres) (Un-Ms) were compared to those of the microspheres washed with a sodium carbonate solution in order to remove the surface drug (treated microspheres) (T-Ms). The results indicate that the removal of the surface drug did not induce any change in the size of the microspheres whereas the morphology of the smallest T-Ms appeared to be modified. The release profiles of both Un-Ms and T-Ms resulted in biphasic patterns. The initial burst effect (first release phase) of the T-Ms was lower than that of the Un-Ms. The rate of the second release phase did not change for the microspheres with the biggest size but increased for the smallest microspheres probably owing to the modification of the matrix porosity. PMID- 10675703 TI - Is bisacodyl absorbed at all from suppositories in man? AB - A HPLC procedure was developed to determine free BHPM in human plasma and urine after prior deconjugation of its glucuronides with glucuronidase. A single dose administration of a 10 mg bisacodyl suppository from Glaxo Wellcome, Poznan (Poland) to 16 volunteers each resulted in its low active metabolite (BHPM) plasma levels (10-55 microgram l(-1)) according to general assumptions. Its prompt laxative effect appeared within 56.6+/-10.8 min. The calculated serum half life time of BHPM glucuronide excretion in urine was approximately 7.32+/-0.99 h. BHPM was excreted in urine in only 3. 36+/-0.52% if compared with the above bisacodyl rectal dose administered. Any relationship between BHPM plasma and/or urine levels and its laxative action does not occur. These results confirm the thesis that the laxative action of bisacodyl suppositories is initiated through a direct interaction of the drug in the rectum. PMID- 10675704 TI - Kinetics of base catalyzed racemization of ibuprofen enantiomers. AB - The kinetics of base catalyzed racemization of ibuprofen enantiomers has been studied in DMSO-water mixed medium. The dynamic equilibrium rate of keto-enol tautomerism leading to racemization of ibuprofen enantiomers, is proportional to the concentrations of base catalyst and substrate. A kinetic model capable of predicting the time course of racemization, under different base and substrate concentrations, is established and experimentally verified. PMID- 10675705 TI - Preformulation studies and characterization of the physicochemical properties of amorphous polymers using artificial neural networks. AB - The utility of artificial neural networks (ANNs) as a preformulation tool to determine the physicochemical properties of amorphous polymers such as the hydration characteristics, glass transition temperatures and rheological properties was investigated. The neural network simulator, CAD/Chem, based on the delta back-propagation paradigm was used for this study. The ANNs software was trained with sets of experimental data consisting of different polymer blends with known water-uptake profiles, glass transition temperatures and viscosity values. A set of similar data, not initially exposed to the ANNs was used to validate the ability of the ANNs to recognize patterns. The results of this investigation indicate that the ANNs accurately predicted the water-uptake, glass transition temperatures and viscosities of different amorphous polymers and their physical blends with a low % error (0-8%) of prediction. The ANNs also showed good correlation between the water-uptake and changes in the glass transition temperatures of the polymers. This study demonstrated the potential of the ANNs as a preformulation tool to evaluate the characteristics of amorphous polymers. This is particularly relevant when designing sustained release formulations that require the use of a fast hydrating polymer matrix. PMID- 10675706 TI - Viscosity prediction of lipophilic semisolid emulsion systems by neural network modelling. AB - Previously published data (Gasperlin et al., 1998) on viscoelastic behaviour of lipophilic semisolid emulsion systems and the prediction of their physical stability by neural network modelling are analysed in further detail. Most attention has been paid to viscosity, which with storage (G') and loss modulus (G"), is one of the most important rheological parameters influenced by structure. Complex dynamic viscosity (eta*) was measured by oscillatory rheometry. The viscosity dependence of the lipophilic semisolid emulsions on the ratio of the particular components was defined by the neural network (error back propagation algorithm), linear and incomplete polynomial models of higher orders. Polynomial models were used to complement the neural network model and to determine the relationship between variables. Since the viscosity was expressed in the whole measured frequency range, modelling was more complex and indirect modelling was introduced. The determined models were tested and the results confirm their usefulness for the explanation and prediction of the rheological characteristics of emulsion systems. The trained and tested neural network model proved to be a highly effective and applicable tool for predicting the viscosity of a lipophilic semisolid emulsion system of given composition. PMID- 10675707 TI - Study on glycolic acid delivery by liposomes and microspheres. AB - Glycolic acid is used in many cosmetic products as exfoliant and moisturizer. Unfortunately, the greater glycolic acid cosmetic benefits the greater is the potential for skin irritation as far as burning. The aim of this work was to investigate the feasibility of topical controlled delivery systems loading glycolic acid in order to optimize the acid cosmetic properties lowering its side effects. For this purpose different types of microparticulate systems have been evaluated: liposomes, liposomes modified by chitosan addition and chitosan microspheres. Liposomes, composed of phosphatidylcholine and cholesterol (1:1 molar ratio) and with different glycolic acid/lipid molar ratio, were prepared by reverse phase evaporation method. Two types of interaction between liposomes and chitosan were investigated: chitosan addition into lipidic bilayer during liposome preparation and coating of already formed liposomes with chitosan. Glycolic acid loaded chitosan microspheres were prepared by the dry-in-oil emulsion method. The microparticulate systems were morphologically characterized by electron microscopy and particle size analysis. In vitro dissolution tests were performed to evaluate the feasibility of microparticulate systems in modulating glycolic acid release. The results obtained show that liposomes are always suitable to modulate glycolic acid release and that the best condition to achieve this control is obtained by the liposomal systems in which glycolic acid/lipid molar ratio is 5:1. Further significant release control is obtained by addition of chitosan into the liposomes, while chitosan microspheres are not able to control glycolic acid release even after crosslinking. PMID- 10675708 TI - Oestradiol skin delivery from ultradeformable liposomes: refinement of surfactant concentration. AB - The aims of this study were to refine ultradeformable liposomes for oestradiol skin delivery and to evaluate Span 80 and Tween 80 as edge activators compared with sodium cholate. Vesicles containing phosphatidylcholine (PC) mixed with edge activators and oestradiol were prepared. Entrapment efficiency and vesicle size were determined. Interactions between activators and vesicles were investigated using differential scanning calorimetry. Transepidermal permeation of oestradiol from vesicles was studied compared to saturated aqueous control in vitro. The maximum flux (J(max)) and its time (T(max)) were calculated from the flux curves and skin deposition was assessed. The compositions of refined formulations were predicted, liposomes prepared, and tested against control. Entrapment efficiency depended on PC concentration with some contribution from sodium cholate and Tween 80. Vesicle sizes ranged from 124 to 135 nm. Edge activators interacted with lipid bilayers and disrupted packing. The refined edge activator concentrations in PC vesicles were 14.0, 13.3 and 15.5% w/w for sodium cholate, Span 80 and Tween 80, respectively; they increased J(max) by 18, 16 and 15-fold and skin deposition by 8, 7 and 8-fold compared with control. Ultradeformable vesicles thus improved skin delivery of oestradiol compared to control and Span 80 and Tween 80 were equivalent to sodium cholate as edge-activators. PMID- 10675709 TI - Effect of formulation and processing variables on the characteristics of microspheres for water-soluble drugs prepared by w/o/o double emulsion solvent diffusion method. AB - 80% except for acetaminophen, due to its lower solubility in water and higher solubility in corn oil. The release profile of the drug was pH dependent. In acidic medium, the release rate was much slower, however, the drug was released quickly at pH 7.4. Tacrine showed unexpected release profiles, probably due to ionic interaction with polymer matrix and the shell structure and the highest release rate was obtained at pH 2.0. The prepared microspheres had a sponge-like inner structure with or without central hollow core and the surface was dense with no apparent pores. PMID- 10675710 TI - The degradation of N,N',N"-triethylenephosphoramide in aqueous solutions: a qualitative and kinetic study. AB - The degradation of N,N',N"-triethylenephosphoramide (TEPA) in aqueous solutions has been investigated over a pH range of 3-14. Samples were analyzed using a gas chromatographic system with nitrogen/phosphorus selective detection. The degradation kinetics were studied as function of pH, sodium chloride concentration and temperature. The degradation of TEPA in buffers follows pseudo first order kinetics. The logk(obs)8 the methoxy derivative of TEPA was formed, as a consequence of the applied procedure. PMID- 10675711 TI - Pulmonary deposition of lactose carriers used in inhalation powders. AB - Dry powder dosage forms are generally formulated by mixing the micronized drug particles with the larger carrier particles. Lactose is a commonly used carrier. Carriers enhance the flowability of powder mixtures and therefore enable low dosing of active substances. During inhalation, the drug particles are dispersed from the surface of carrier particles. The aim of this study was to compare how different qualities of 99mTc-labelled lactose carrier systems deposit in the lungs. The sizes of the labelled and unlabelled alpha-lactose monohydrate particles were compared by using a laser diffraction method. Distribution of radiolabel between different particle size fractions was determined using the Andersen cascade impactor. The in vivo depositions of lactose carrier systems were investigated in ten healthy men using the technique of gammascintigraphy. In addition, redispersion of budesonide from the carrier materials was evaluated by using the Andersen cascade impactor. According to the validation data the particle size of the lactose carriers remained unchanged during the labelling process. Low pulmonary deposition varying between 2.5 and 3.3% was detected. Only a small amount of lactose was deposited in the lungs, thus pulmonary deposition is not a limiting factor for lactose selection. According to in vitro redispersion data the fine particle fraction of the delivered dose in the impactor varied between 10.3 and 26.0%. Thus, the redispersion of the budesonide particles can be altered by the properties of the carrier system. PMID- 10675712 TI - Effect of vehicles and pressure sensitive adhesives on the permeation of tacrine across hairless mouse skin. AB - The purpose of this study was to investigate the feasibility of developing transdermal drug delivery (TDD) system for tacrine used for treating the symptoms of Alzheimer's disease. The effects of various vehicles on the percutaneous absorption of tacrine in solution formulation and in pressure sensitive adhesive (PSA) matrix across the hairless mouse skin were evaluated using flow-through diffusion cell system at 37 degrees C. The permeation profiles of tacrine from solutions were different depending on vehicles used. The flux of tacrine increased significantly as its concentration in the solutions increased. The permeation rate of tacrine was higher in acrylic adhesives with hydroxyl functional group and without functional group than in polyisobutylene adhesive matrix. Incorporation of vehicles into the acrylic adhesive matrix significantly enhanced the permeation rate and shortened the lag time of tacrine. The maximum flux obtained from pressure sensitive adhesive matrix seemed to be high enough to obtain therapeutic effect. PMID- 10675713 TI - Gentamicin encapsulation in PLA/PLGA microspheres in view of treating Brucella infections. AB - In view of treating intracellular Brucella infections, microspheres made of poly(lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) were developed as delivery system for the cationic and highly hydrophilic antibiotic gentamicin sulphate. Drug microencapsulation by spray drying yielded microspheres with regular morphology, an average particle size of approximately 3 micrometer and encapsulation efficiencies of up to 45%. Different copolymers of similar molecular weights gave varying encapsulation efficiencies and particle size distributions. The encapsulation efficiency generally increased with polymer hydrophilicity, except for the hydrophilic copolymer PLGA50:50H carrying carboxylic end groups. Encapsulation also depended on the pH value of the aqueous drug solution to be encapsulated. Moreover, increasing nominal gentamicin sulphate loading yielded lower efficiencies. For comparison, some formulations were also prepared by a (W(1)/O)W(2)-solvent evaporation method, which yielded lower encapsulation efficiencies, in the order of 13%. Finally, drug bioactivity was found to remain intact after microencapsulation, MS storage and MS incubation in aqueous medium. The results suggest that PLA/PLGA microspheres prepared by spray drying may be an appropriate delivery system for gentamicin sulphate to be used in the treatment of intracellular Brucella infections. PMID- 10675714 TI - Predictions of coronary artery stenosis by artificial neural network. AB - Data from angiography patient records comprised 14 input variables of a neural network. Outcomes (coronary artery stenosis or none) formed both supervisory and output variables. The network was trained by backpropagation on 332 records, optimized on 331 subsequent records, and tested on final 100 records. If 0.40 was chosen as the output distinguishing stenosis from no stenosis, 81 patients who had stenosis would have been identified, while 9 of 19 patients who did not have stenosis might have been spared angiography. The results demonstrated that artificial neural networks could identify some patients who do not need coronary angiography. PMID- 10675715 TI - Generating concise and accurate classification rules for breast cancer diagnosis. AB - In our previous work, we have presented an algorithm that extracts classification rules from trained neural networks and discussed its application to breast cancer diagnosis. In this paper, we describe how the accuracy of the networks and the accuracy of the rules extracted from them can be improved by a simple pre processing of the data. Data pre-processing involves selecting the relevant input attributes and removing those samples with missing attribute values. The rules generated by our neural network rule extraction algorithm are more concise and accurate than those generated by other rule generating methods reported in the literature. PMID- 10675716 TI - Planning treatment of ischemic heart disease with partially observable Markov decision processes. AB - Diagnosis of a disease and its treatment are not separate, one-shot activities. Instead, they are very often dependent and interleaved over time. This is mostly due to uncertainty about the underlying disease, uncertainty associated with the response of a patient to the treatment and varying cost of different diagnostic (investigative) and treatment procedures. The framework of partially observable Markov decision processes (POMDPs) developed and used in the operations research, control theory and artificial intelligence communities is particularly suitable for modeling such a complex decision process. In this paper, we show how the POMDP framework can be used to model and solve the problem of the management of patients with ischemic heart disease (IHD), and demonstrate the modeling advantages of the framework over standard decision formalisms. PMID- 10675718 TI - Foreword. PMID- 10675717 TI - Automatic analysis of signals with symbolic content. AB - This paper presents a set of methods for helping in the analysis of signals with particular features that admit a symbolic description. The methodology is based on a general discrete model for a symbolic processing subsystem, which is fuzzyfied by means of a fuzzy inference system. In this framework a number of design problems have been approached. The curse of dimensionality problem and the specification of adequate membership functions are the main ones. In addition, other strategies, which make the design process simpler and more robust, are introduced. Their goals are automating the production of the rule base of the fuzzy system and composing complex systems from simpler subsystems under symbolic constrains. These techniques are applied to the analysis of wakefulness episodes in the sleep EEG. In order to solve the practical difficulty of finding remarkable situations from the outputs of the symbolic subsystems an unsupervised adaptive learning technique (FART network) has been applied. PMID- 10675719 TI - Unraveling the role of proteases in cancer. AB - Investigators have been studying the expression and activity of proteases in the final steps of tumor progression, invasion and metastasis, for the past 30 years. Recent studies, however, indicate that proteases are involved earlier in progression, e.g., in tumor growth both at the primary and metastatic sites. Extracellular proteases may co-operatively influence matrix degradation and tumor cell invasion through proteolytic cascades, with individual proteases having distinct roles in tumor growth, invasion, migration and angiogenesis. In this review, we use cathepsin B as an example to examine the involvement of proteases in tumor progression and metastasis. We discuss the effect of interactions among tumor cells, stromal cells, and the extracellular matrix on the regulation of protease expression. Further elucidation of the role of proteases in cancer will allow us to design more effective inhibitors and novel protease-based drugs for clinical use. PMID- 10675720 TI - An overview of collagenase-3 expression in malignant tumors and analysis of its potential value as a target in antitumor therapies. AB - Collagenase-3 (MMP-13) is a member of the matrix metalloproteinase family of endopeptidases that is characterized by a potent degrading activity against a wide spectrum of substrates. This enzyme was first detected in breast carcinomas but it is also overexpressed in a variety of malignant tumors including head and neck carcinomas, chondrosarcomas, skin carcinomas, and carcinomas of the female genital tract. Clinical studies have revealed that in all these tumors collagenase-3 expression is associated with invasive and metastatic tumors. Analysis of the molecular mechanisms underlying its marked overexpression in malignant tumors has allowed to identify different cytokines, growth factors and tumor promoters with ability to up-regulate collagenase-3 expression in tumor cells, or in stromal fibroblasts surrounding epithelial tumor cells. The first strategies designed to target this enzyme are being developed, and are mainly directed to prepare synthetic inhibitors with ability to selectively block the collagenase-3 proteolytic activity. Alternatively, inhibitors of the signal transduction pathways mediating the expression of this enzyme by tumor cells may also be useful for collagenase-3 targeting. These studies together with those performed on other enzymes associated with tumor processes may lead to the development of novel therapeutic strategies to control the progression and metastatic capacity of neoplastic cells. PMID- 10675721 TI - Cathepsin D in breast cancer: mechanisms and clinical applications, a 1999 overview. AB - A short review of the literature first confirms the clinical value of cathepsin D as a prognostic marker in breast cancer, when using well standardized assays. We then summarize results of studies, mostly performed in our laboratory, aimed at understanding the effect of cathepsin D overexpression on metastasis and the molecular mechanisms involved. Cathepsin D-cDNA transfection increases tumor cell proliferation in vitro and the metastatic potential of 3Y1-Ad12 embryonic rat tumorigenic cells when injected in vivo into nude mice. The mechanism by which cathepsin D increases the incidence of clinical metastasis involves increased cell growth and decreased contact inhibition rather than escape of cancer cells through the basement membrane. Different mechanisms are considered to explain this mitogenic activity. Cathepsin D could act as a protease following its activation at an acidic pH, or as a ligand of different membrane receptors at a more neutral pH. In this case cathepsin D can displace IGFII from the mannose-6 phosphate/IGFII receptor to the IGFI receptor or activate another membrane receptor to be identified. The nature of the mechanisms involved in vivo may depend on the micro environment of the tumor cells. These studies should guide in the development of new therapies aimed at inhibiting the deleterious effect of overexpressed cathepsin D. PMID- 10675722 TI - Proteases in gastrointestinal neoplastic diseases. AB - Cysteine and serine proteases are involved in cancer invasion and metastasis. In the past few years we investigated the tissue levels of these proteases in gastric cancer (GC), gastric precancerous changes (CAG), colorectal cancer (CRC) and the plasma and serum levels of proteases in several gastrointestinal tumours, using ELISA methods. Significantly higher antigen levels were found not only in GC tissue but also in CAG with respect to the levels found normal tissue; with respect to CAG, patients with dysplasia had higher levels than patients without dysplasia. The same findings were obtained in CRC. In general protease levels correlated with the major histomorphological parameters, such as grading and histotype in GC as well as in CRC. Tissue protease levels had a strong prognostic impact in GC, in which UPA was singled out by multivariate analysis as the major prognostic factor, and CRC. The plasma levels of urokinase-type plasminogen activator (UPA) and the serum levels of cathepsin B were significantly increased in patients with gastrointestinal tumours. In conclusions, cysteine and serine proteases may have a part not only in GC and CRC invasion and metastasis, but also in the progression of gastric precancerous changes into cancer. They are strong prognostic factors in GC and CRC. These proteases may also have a role as tumour markers in the early diagnosis of gastrointestinal tract tumours. PMID- 10675723 TI - Bacterial toxins with intracellular protease activity. AB - The recent determination of their primary sequence has lead to the discovery of the metallo-proteolytic activity of the bacterial toxins responsible for tetanus, botulism and anthrax. The protease domain of these toxins enters into the cytosol where it displays a zinc-dependent endopeptidase activity of remarkable specificity. Tetanus neurotoxin and botulinum neurotoxins type B, D, F and G cleave VAMP, an integral protein of the neurotransmitter containing synaptic vesicles. Botulinum neurotoxins type A and E cleave SNAP-25, while the type C neurotoxin cleaves both SNAP-25 and syntaxin, two proteins located on the cytosolic face of the presynaptic membrane. Such specific proteolysis leads to an impaired function of the neuroexocytosis machinery with blockade of neurotransmitter release and consequent paralysis. The lethal factor of Bacillus anthracis is specific for the MAPkinase-kinases which are cleaved within their amino terminus. In this case, however, such specific biochemical lesion could not be correlated with the pathogenesis of anthrax. The recently determined sequence of the vacuolating cytotoxin of Helicobacter pylori contains within its amino terminal domain elements related to serine-proteases, but such an activity as well as its cytosolic target remains to be detected. PMID- 10675724 TI - Proteases in the evaluation of pancreatic function and pancreatic disease. AB - This paper reviews the role of pancreatic proteases (focusing upon trypsin, chymotrypsin and elastase) in the diagnosis and management of chronic pancreatic insufficiency (CPI), emphasizing advances over the last 5 years. Some important novel aspects of these enzymes in acute pancreatitis are also described, including their role in diagnosis and their interaction with cholecystokinin in the pathogenesis of the disease. The recent interest in these enzymes as agents promoting the spread of cancer in animals and human subjects is also described. A hierarchical approach has been taken to explore the advantages and limitations of tests in different source materials: serum, feces, duodenal aspirate, and non invasive pancreatic function tests. The practical usefulness of fecal elastase-1 and of fecal chymotrypsin concentrations in diagnosis and management of CPI, respectively, is one of the major lessons to be learned from analysis of the recent literature, and forms the principal message of this review. PMID- 10675725 TI - Proteinases in bone resorption: obvious and less obvious roles. AB - Bone resorption is critical for the development and the maintenance of the skeleton, and improper regulation of bone resorption leads to pathological situations. Proteinases are necessary for this process. In this review, we show that this need of proteinases is not only because they are required for the solubilization of bone matrix, but also because they are key components of the mechanism that determines where and when bone resorption will be initiated. Moreover, there are indications that proteinases may also determine whether resorption will be followed by bone formation. Some of the proteinases involved in these different steps of the resorption processes were recently identified, as for instance cathepsin K, MMP-9 (gelatinase B), and interstitial collagenase. However, there is also increasing evidence showing that the critical proteinase(s) may vary depending on the bone type or on other factors. PMID- 10675726 TI - Tissue factor in human coronary atherosclerotic plaques. AB - The rupture or fissuring of a coronary atherosclerotic plaque and subsequent thrombosis is considered the key event in the pathogenesis of unstable angina and myocardial infarction. Although plaque disruption frequently occurs during the evolution of atherosclerosis, only a minority of ruptured plaques develop thrombosis. The content and procoagulant activity of tissue factor in human coronary atherosclerotic plaques varies widely, and different studies confirm that it is higher in the plaques extracted from patients with unstable angina, myocardial infarction or histologic/angiographic evidence of coronary thrombosis than in those taken from patients with stable angina or uncomplicated coronary lesions. Variations in tissue factor content and activity may be responsible for the different thrombotic responses to human coronary atherosclerotic plaque rupture. PMID- 10675727 TI - Inhibition of cytosolic phospholipase A(2) attenuates activation of mitogen activated protein kinases in human monocytic cells. AB - Eicosanoids and platelet-activating factor generated upon activation of cytosolic phospholipase A(2) enhance activity of transcription factors and synthesis of proinflammatory cytokines. Here, we show that selective inhibitors and antisense oligonucleotides against this enzyme suppressed expression of the interleukin 1beta gene at the level of transcription and promoter activation in human monocytic cell lines. This inhibitory effect was due to failure of activation of mitogen-activated protein kinases (MAPK) through phosphorylation by upstream mitogen-activated protein kinase kinases (MKK). Consequently, phosphorylation and degradation of inhibitor-kappaBalpha (I-kappaBalpha) and subsequent cytoplasmic mobilization, DNA-binding and the transactivating potential of nuclear factor kappaB (NF-kB), nuclear factor-interleukin-6 (NF-IL6), activation protein-1 (AP 1) and signal-transducer-and-activator-of-transcription-1 (STAT-1) were impaired. It is concluded, that lipid mediators promote activation of MAPKs, which in turn lead to phosphorylation and liberation of active transcription factors. Since inhibition of cytosolic phospholipase A(2) ameliorates inflammation in vivo, this potency may reside in interference with the MAPK pathway. PMID- 10675728 TI - Nifedipine does not induce but rather prevents apoptosis in cardiomyocytes. AB - The potential of Ca(2+) channel antagonists, particularly nifedipine, to cause apoptotic cell death has been controversial and is of considerable importance for cardiomyocytes as loss of these cells is an important component of the pathophysiology leading to heart failure. To examine the hypothesis that nifedipine induces cell death and modulates calcium-induced apoptosis, cardiomyocytes in culture from embryonic chick hearts, that readily manifest apoptosis, were studied. Apoptosis was evaluated by fluorescent activated cell sorting (FACS) analysis and by quantitative analysis of DNA fragmentation by an enzyme-linked immunosorbent assay (ELISA) specific for histone-associated DNA fragments of mono- and oligonucleosome size. Cell death was evaluated by using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. Cardiomyocytes were treated with various concentrations of nifedipine over a concentration range relevant to serum concentrations in man. Nifedipine, 1 to 100 microM, did not produce cell death in cardiomyocytes. There was no evidence of apoptosis on FACS analysis of cardiomyocytes stained with fluoresceine diacetate or propidum iodide (PI). Neither was there any evidence of apoptotic nuclei on PI staining of permeabilized cardiomyocytes treated with nifedipine. In contrast, DNA fragmentation consistent with apoptosis was induced in a significant (P<0.05) concentration-dependent manner, by increases in extracellular Ca(2+) concentration ([Ca(2+)](o)). Importantly, nifedipine reduced DNA fragmentation produced by increased [Ca(2+)](o). Furthermore, nifedipine blocked calcium induced cell death as high [Ca(2+)](o) significantly (P<0. 05) reduced cell viability. These data indicate that nifedipine does not induce apoptosis in cardiomyocytes rather apoptosis in cardiomyocytes is under regulatory control by Ca(2+) and nifedipine can antagonize Ca(2+)-mediated apoptotic cell death. PMID- 10675729 TI - Myelopoietic response in tumour-bearing mice by an aggregated polymer isolated from Aspergillus oryzae. AB - The effects of magnesium ammonium phospholinoleate-palmitoleate anhydride (MAPA), a proteic aggregated polymer isolated from Aspergillus oryzae, on the growth and differentiation of granulocyte-macrophage progenitor cells (colony-forming unit granulocyte-macrophage [CFU-GM]) in normal and Ehrlich ascites tumour-bearing mice were studied. Myelosuppression concomitant with increased numbers of spleen CFU-GM was observed in tumour-bearing mice. Treatment of these animals with MAPA (0.5-10 mg/kg) stimulated marrow myelopoiesis in a dose-dependent manner and reduced spleen colony formation. No changes were observed in total and differential marrow cell counts. The dose of 5.0 mg/kg MAPA, given prior or after tumour inoculation, was the optimal biologically active dose in tumour-bearing mice and this dose schedule also stimulated myelopoiesis in normal mice. MAPA significantly enhanced survival and concurrently reduced tumour growth in the peritoneal cavity. We propose that the modulatory effect of MAPA on the myelopoietic response may be related to its antitumour activity as a possible mechanism for regulation of granulocyte-macrophage production and expression of functional activities. PMID- 10675730 TI - cAMP-Dependent potentiation of the Ca(2+)-activated release of the anionic fluorescent dye, calcein, from rat parotid acinar cells. AB - A recent study indicates that elevation of [Ca(2+)](i) enhances the release of calcein, an anionic fluorescent dye, from isolated exocrine acinar cells, so cytoplasmic calcein is useful for monitoring the secretion of organic anions. In this study, we investigated the effect of cAMP on the calcein release evoked by elevation of [Ca(2+)](i). Isoproterenol, forskolin and dibutyryl cyclic AMP (dbcAMP) did not induce the release of calcein from isolated parotid acinar cells, but they potentiated the carbachol-induced release of calcein. Although cytoplasmic calcein is released through an increase in [Ca(2+)](i), isoproterenol potentiated the carbachol-induced release of calcein without affecting the increase in [Ca(2+)](i) evoked by a high concentration of carbachol (10(-6) M). Charybdotoxin, a K(+) channel blocker, inhibited both the carbachol-induced release and the potentiation by isoproterenol. However, the calcein permeation pathways mediating the carbachol-induced release and the isoproterenol potentiated release exhibited distinct sensitivities to anion channel blockers. Our results indicate that the calcein release induced by carbachol is potentiated through an increase in intracellular levels of cAMP. Although both the Ca(2+) activated release and the cAMP-potentiated release may be coupled to Ca(2+) activated K(+) efflux, increases in both [Ca(2+)](i) and [cAMP](i) may activate the calcein conduction pathway which is not activated by an increase in [Ca(2+)](i) alone. PMID- 10675731 TI - Binding properties of [3H]gacyclidine in the rat central nervous system. AB - Gacyclidine (1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine), the racemate of (+) and (-)-GK11, exhibits potent neuroprotective properties due to its antagonism at the NMDA receptor. In its tritiated form, gacyclidine showed a binding distribution similar to that of NMDA receptors in the rat brain. With membrane preparations, the (-)-enantiomer of gacyclidine exhibited an affinity similar to that of MK-801 (dizocilpine, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a, d]cyclohepten-5,10-imine) in the low nanomolar range, while the (+)-enantiomer was about 10 times less potent. Gacyclidine affinity was lower in the cerebellum than in the forebrain or the spinal cord. In this latter region and in the cerebellum, two binding sites were evidenced, one of which was a low-affinity site insensitive to MK-801. In all regions, PRE-084 (2-(4-morpholino)ethyl-1 phenylcyclohexane-1-carboxylate), a sigma receptor ligand, had no effect on [3H]gacyclidine binding. PMID- 10675733 TI - Roles of adenosine A(1) and A(2A) receptors in the expression and development of methamphetamine-induced sensitization. AB - We studied the effects of adenosine A(1) and A(2A) receptor agonists on the expression and development of methamphetamine-induced sensitization in rats. When animals were treated with the adenosine A(1) receptor agonist, N(6) cyclohexyladenosine (CHA), along with methamphetamine every 3 days with a total of five administrations, the augmentation of hyperlocomotion by methamphetamine re-administration after 7-day withdrawal (methamphetamine challenge administration) was not inhibited. However, when the adenosine A(2A) receptor agonist, 2-p-(2-carboxyethyl) phenethyl-amino-5'-N-ethylcarboxy-amide adenosine (CGS21680), was administered according to the same schedule, the augmentation was significantly inhibited. On the other hand, when CHA or CGS21680 was administered 30 min before methamphetamine challenge, both drugs dose-dependently inhibited the augmentation of hyperlocomotion. These results suggested that both adenosine A(1) and A(2A) receptors play important roles in the expression of methamphetamine-induced sensitization, and that adenosine A(2A) receptors do so in the development of this sensitization. PMID- 10675732 TI - delta-Opioid receptor agonists produce antinociception and [35S]GTPgammaS binding in mu receptor knockout mice. AB - We examined the effects of [D-Pen(2),D-Pen(5)]enkephalin (DPDPE), [D Ala(2),Glu(4)]deltorphin (DELT), and (+)-4-[(alphaR)-alpha((2S, 5R)-4-Allyl-2,5 dimethyl-1-piperazinyl)-3-methoxybenzyl]-N, N-diethylbenzamide (SNC80) on [35S]GTPgammaS binding in brain membranes prepared from micro-opioid receptor knockout (-/-) mice. The potency and maximal response (E(max)) of these agonists were unchanged compared to control mice. In contrast, while the potency of [D Pen(2),pCl-Phe(4),D-Pen(5)]enkephalin (pCl-DPDPE) was not significantly different, the E(max) was reduced as compared to controls. In the tail-flick test, intracerebroventricular (i.c.v.) or intrathecal (i.th.) DELT produced antinociceptive effects in -/- mice with potency that did not differ significantly from controls. In contrast, the antinociceptive potency of i.c.v. and i.th. DPDPE was displaced to the right by 4- and 9-fold in -/- compared to control mice, respectively. Reduced DPDPE antinociceptive potency in -/- mice, taken together with reduced DPDPE- and pCl-DPDPE- stimulated G protein activity in membranes prepared from -/- mice, demonstrate that these agonists require mu opioid receptors for full activity. However, because DELT mediated G protein activation and antinociception were both comparable between -/- and wild type mice, we conclude that the mu-opioid receptor is not a critical component of delta-opioid receptor function. PMID- 10675734 TI - Effect of somatostatin on resistance and on capacitance rabbit isolated arteries. AB - The effects of somatostatin, a tetradecapeptide isolated from hypothalamus extracts, were studied on the vascular reactivity of aorta and mesenteric arteries isolated from rabbits. We also investigated whether or not Ca(2+) movements were implicated in these effects. Rabbit aorta and mesenteric (fifth branch) arteries were isolated, cleaned off, and mounted in an organ bath containing Godfraind solution or physiological saline solution (PSS), respectively. Somatostatin (10(-8)-10(-4) M) produced a concentration-dependent inhibition of the contractile responses induced by high K(+) (80 mM) or noradrenaline (10(-6) M in aorta or 10(-4) M in mesenteric arteries) in both arteries studied. The inhibitory effect of somatostatin was greater in mesenteric resistance vessels (IC(50) 3.1+/-2.3x10(-5) M, and 5.2+/-4.8x10(-8) M with KCl and noradrenaline, respectively). Contractile responses produced by the addition of Ca(2+) (1-5 mM) to Ca(2+)-free high K(+) solution were also concentration dependently inhibited by somatostatin in aorta. Furthermore, somatostatin decreased noradrenaline-induced contraction attributed to intracellular Ca(2+) release in aorta, and inhibited 45Ca(2+) uptake stimulated by high K(+) or by noradrenaline. However, it did not modify 45Ca(2+) uptake in resting mesenteric resistance arteries. Taken together, these results suggest that somatostatin exerts an inhibitory effect on vascular contractions induced by some stimulating agents in different arteries isolated from rabbits, being more potent in mesenteric arteries. PMID- 10675735 TI - Effects of metoprolol and ramipril on action potentials after myocardial infarction in rats. AB - The effects of chronic treatment with the beta-adrenoceptor antagonist metoprolol, the angiotensin converting enzyme inhibitor ramipril, their combination, or placebo on action potential configuration 6 weeks after myocardial infarction in rats were studied. Action potentials were measured in isolated left ventricular posterior papillary muscles and compared with action potentials from a sham operated group without infarction. After infarction, the action potential amplitude was reduced and this phenomenon was partially reversed by metoprolol- and ramipril-treatment. Prolonged repolarisation after infarction compared to sham operated animals was additionally delayed after metoprolol treatment. Thus, metoprolol extends the refractory period, which may counteract tachyarrhythmia. PMID- 10675736 TI - Prostaglandin-release impairment in the bladder epithelium of streptozotocin induced diabetic rats. AB - Isolated epithelial layer preparations were obtained from urinary bladders of 4 week streptozotocin-diabetic rats and used for endogenous prostaglandins E(2) and F(2alpha) determination. Tissues were incubated in modified Krebs solution under basal conditions, or in the presence of either indomethacin (5x10(-7) M), ATP (10(-5) and 10(-3) M) or bradykinin (10(-7) and 10(-5) M), and samples of incubation medium were collected at 15 and 30 min. In the presence of indomethacin, the release of prostaglandins in the incubation medium was under the detection limit of the enzyme immunoassay (EIA). The epithelium from diabetic rat urinary bladders was thicker and heavier and the absolute amount of endogenous prostaglandins E(2) and F(2alpha) was higher than for control animals, but when prostaglandin production was expressed as a fraction of tissue weight, it was reduced in diabetic epithelium. ATP and bradykinin has significantly increased the endogenous release of both prostaglandins from the epithelium when compared with the release under basal conditions. This increase was time dependent and was higher in diabetic than in control tissues. ATP evoked a phasic and tonic contraction in bladder strips that was abolished by epithelium removal. Concentration-response curves for ATP did not differ among groups. Bradykinin evoked a long-lasting tonic contraction that was reduced significantly by epithelium removal in diabetic rat bladders only. Concentration-response curves for prostaglandin E(2) and F(2alpha) in diabetic rat bladder differed significantly from that in controls and epithelium removal did not alter these responses. It is suggested that bradykinin receptors and P2X nucleotide receptors already found in the smooth muscle detrusor might be present in the epithelial layer of the bladder. The prostaglandin-release impairment observed in this study might be responsible, in part, for bladder abnormalities observed in pathological conditions, such as diabetes. PMID- 10675737 TI - Urinary nitrate excretion in cholesterol-fed rabbits: effect of a chronic treatment by N-iminoethyl-L-lysine, a selective inhibitor of inducible nitric oxide synthase. AB - To evaluate the influence of atherosclerosis on the global production of NO, we studied the effect of a 0.3% cholesterol-enriched diet on urinary nitrate excretion in rabbits during 69 weeks. To examine whether the inducible nitric oxide synthase (iNOS) present in atherosclerotic lesions could participate in NO excretion, hypercholesterolemic rabbits were treated chronically with the selective iNOS inhibitor, N-iminoethyl-L-lysine (L-NIL; 5 mg/kg/day). Urinary nitrate excretion was higher in hypercholesterolemic than in control rabbits throughout the study period and decreased progressively with time in both groups; L-NIL had no significant effect on urinary nitrate excretion. These data illustrate that systemic NO production is enhanced in hypercholesterolemia and that iNOS, present within the plaque, might not participate in this enhanced NO production. PMID- 10675738 TI - Site-specific lesion formation, inflammation and inducible nitric oxide synthase expression by indomethacin in the rat intestine. AB - The involvement of nitric oxide (NO) formed by the inducible isoform of NO synthase (iNOS) has been investigated in the development of rat intestinal lesions following indomethacin administration. Over a 72-h period, indomethacin (10 mg kg(-1), s.c.) provoked a time-dependent increase in expression of iNOS (assessed by the conversion of radiolabelled L-arginine to citrulline) and enhancement of vascular leakage of radiolabelled human serum albumin in the jejunum which commenced 18 h after indomethacin. Similar effects were not observed in the ileum, colon or caecum. In addition, macroscopic lesions were detectable and myeloperoxidase activity (an index of neutrophil recruitment) were increased in the rat jejunum 18-24 h after indomethacin, but remained at basal levels in the ileum and colon. These findings suggest that indomethacin provokes a site-selective expression of iNOS in the rat jejunum which correlates with lesion formation and vascular leakage, whereas both the ileum and colon are spared. PMID- 10675739 TI - Systematics of malaria vectors with particular reference to the Anopheles punctulatus group. AB - The appearance of groups and complexes of closely related cryptic or sibling species in many of the anopheline taxa has impeded studies on malaria transmission and the evaluation of control strategies which have relied on morphological characters to identify the vector species involved. The advantages of morphological identification are low cost, speed and simplicity, which allow large numbers of specimens to be processed rapidly in the field. The need for accurate identification is crucial, as time and money may be wasted in studying and controlling species of no medical importance. Various techniques such as cross-mating, chromosome studies and allozyme analysis have been developed to resolve problems of identifying sibling species, though none, as yet, can match the speed and simplicity afforded by morphology markers. The latest of these identification methods comes from advances that have been made in DNA-based technology. Although costly and requiring fairly sophisticated laboratory support, methods such as DNA probe hybridisation and PCR are the quickest and most user-friendly to date. The use of DNA has other advantages in the study of intraspecific differences and in providing characters for phylogenetic studies. This review looks at the development of DNA-based techniques for taxonomic and systematic studies of anopheline mosquitoes. The Anopheles punctulatus group of the southwest Pacific is featured as an example of how this technology has been applied and how it has progressed. PMID- 10675740 TI - A comparative survey of the hydrolytic enzymes of ectoparasitic and free-living mites. AB - Extracts of ectoparasitic mites of birds (Dermanyssus gallinae), sheep (Psoroptes ovis) and plants (Tetranychus urticae) and of free-living mites (Acarus siro) contained acid and alkaline phosphatase, C4 and C8 esterases, lipase, leucine and valine aminopeptidases and a range of glycosidase activities. Dermanyssus gallinae and P. ovis, species highly adapted to an animal parasitic lifestyle, had very similar profiles and contained low activities of glycosidases. In contrast, the polyphagous species A. siro contained moderate to high activities of every glycosidase examined, whereas the phytophagous species, T. urticae, displayed high activities of only beta-galactosidase and beta-glucuronidase. All extracts hydrolysed haemoglobin with optima below pH6, and this hydrolysis was associated with an aspartic proteinase and variable cysteine proteinase activity dependent on species. Inhibitor-labelling with biotinyl-Phe-Ala-FMK revealed the presence of cysteine proteinases with molecular masses of 25-33.5kDa. Each mite species contains the enzymes necessary to complete digestion of the diet in the intracellular lysosomal compartment. The absolute and relative activities of each enzyme varied, and are discussed according to phylogeny and dietary habit. PMID- 10675741 TI - Comparative metal content profiling of parasitic helminths by electron paramagnetic resonance spectrometry: significance for metalloprotein content. AB - Variation in co-ordination geometries of metal ions bound to proteins imposes electronic states different from free (hydrated) ions in solution. Electron paramagnetic resonance spectroscopy has been used to analyse a selection of parasitic helminths for metal content as an initial step to determination of metallo-enzymes in their ES products under immune stress conditions. Characteristic paramagnetic resonance spectroscopy spectra show clear evidence for the presence of iron, copper, and manganese centres and in the selected parasites. The metals ions are identified as protein-bound as distinct from free metal ions present in aqueous solution, and distinguishable from parasite dietary components derived from host sources. Indication is given that superoxide dismutases may, in part, account for the metal ions observed. The use of electron paramagnetic resonance spectroscopy to identify specific protein-bound metals without prior isolation of the suspected protein is here applied. PMID- 10675742 TI - Identification of six Trypanosoma cruzi phylogenetic lineages by random amplified polymorphic DNA and multilocus enzyme electrophoresis. AB - Genetic characterisation of Trypanosoma cruzi variants is of foremost importance, due to considerable genetic and biological heterogeneity in the parasite populations. Two major phylogenetic lineages, each highly heterogeneous, have been previously described within this species. Here we characterised a geographically and ecologically diverse sample of stocks representative of the breadth of the known clonal diversity of each major lineage, using random amplified polymorphic DNA with 20 primers and multilocus enzyme electrophoresis at 22 loci. Molecular hybridisation experiments were performed to control the homology of randomly amplified DNA markers. Both sets of data were highly consistent and supported the existence of two major lineages. Additionally, we found that lineage 2 appeared further partitioned into five sharply delineated phylogenetic clusters, each comprising one of the following reference strains: CanIII cl1 (Z3 reference), M5631 cl5, Esmeraldo cl3 (Z2 reference), CL Brener, and MN cl2. The two first clusters were found mainly in sylvatic environments, whereas the three latter were restricted to domestic transmission cycles and were only collected South to the Amazon Basin. In contrast, lineage 1, which included Miles' Z1 reference strain X10 cl1, was not further subdivided and was encountered across the entire endemic area, in both domestic and sylvatic cycles. Thus, T. cruzi appeared to be subdivided into six discrete typing units, or DTUs, exhibiting distinct geographic and ecological ranges. Reliable diagnostic markers for the two major lineages and the five smaller DTUs of lineage 2 are described, and correspondence with previous classifications of T. cruzi genotypes is given in order to help communication on T. cruzi phylogenetic diversity. PMID- 10675743 TI - Characterisation of the proximal regulatory domain of the Echinococcus granulosus homeodomain-containing gene EgHbx1. AB - In an attempt to understand the molecular basis of the development of Echinococcus granulosus, we have previously isolated several homeobox-containing genes (EgHbx1-5). Here we report the characterisation of the EgHbx1 proximal regulatory domain. EgHbx1 codes for an amino acid sequence presenting 90% identity at the homeodomain level with the Drosophila melanogaster NK1-S59 transcription factor. The proximal regulatory domain sequence and the transcription start site were determined. Electrophoretic mobility shift assays using nuclear extracts from protoscoleces revealed the presence of specific DNA protein complexes. These results constitute a hint and provide new tools to decipher regulatory cascades during E. granulosus development. PMID- 10675745 TI - A common high molecular weight antigen of Babesia bovis isolates from Mexico. AB - Cattle from an area of Mexico endemic with Babesia bovis infections have a dominant antibody response to a 152kDa antigen of the Tamaulipas strain of B. bovis. A mAb termed PB/5, showing a specific reactivity to this 152kDa antigen in Western blots, was identified. The mAb which reacted with the blunt end of B. bovis in an indirect fluorescent antibody test also reacted to a 152kDa antigen in two other isolates (Nuevo Leon and Yucatan), and a 175kDa antigen in the Huasteca B. bovis isolate from Mexico. Polyclonal monospecific sera from a calf inoculated with mAb-affinity purified 152kDa antigen (Tamaulipas strain) identified B. bovis by the indirect fluorescent antibody test and two antigens of B. bovis (65kDa and 152kDa) in Western blot. Since the epitope reacting to the mAb PB/5 is conserved, this antigen provides a basis for developing a diagnostic test or an immunogen. PMID- 10675744 TI - Monoclonal antibody inhibition of Neospora caninum tachyzoite invasion into host cells. AB - Monoclonal antibodies were produced against Neospora caninum tachyzoites to identify antigens which may play a role during invasion of host cells. Confocal laser microscopy showed that most antigens recognised by the mAb were located on the surface, but one mAb, 1A5, reacted to the apical end of the parasite. Some mAbs, which recognised 70, 42 and 36kDa parasite proteins, significantly inhibited the invasion of the parasite in vitro. The mAbs which recognised 42 and 36kDa parasite protein, reacted with Nc-p43 and Nc-p36 expressed by vaccinia virus and Escherichia coli, respectively. These results suggest that a 70kDa protein, Nc-p43 and Nc-p36 are involved in the invasion of the parasite into host cells. PMID- 10675746 TI - Survival of bird schistosomes in mammalian lungs. AB - Bird schistosome cercariae have a low specificity to vertebrate skin and, thus, they are also able to penetrate into mammals. As a consequence, a hypersensitive skin response-cercarial dermatitis-develops. It was thought that the parasites die in the skin soon after penetration. Our results on Trichobilharzia szidati and Bilharziella polonica in the non-specific murine host confirm that some of the penetrating bird schistosomes may fully transform to schistosomula and migrate to the lungs. They persist there for up to 10days post exposure. In a duck, the worms grow and feed rapidly, but in a mouse the lung schistosomula seem to be inhibited in their development. However, TEM results show that there is no damage to the tegument of these larvae and no immune effector cells attack the parasites. These results suggest that the parasite's failure in the murine host might be caused by some immunologically unrelated factors. PMID- 10675747 TI - Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. AB - We have identified a novel 529bp fragment that is repeated 200- to 300-fold in the genome of Toxoplasma gondii. This 529bp fragment was utilised for the development of a very sensitive and specific PCR for diagnostic purposes, and a quantitative competitive-PCR for the evaluation of cyst numbers in the brains of chronically infected mice. The 529bp fragment was found in all 60 strains of T. gondii tested, and it discriminates DNA of T. gondii from that of other parasites. Toxoplasma gondii DNA was detected in amniotic fluid of patients, as well as in various tissues from infected mice. Polymerase chain reaction with the 529bp fragment was more sensitive than with the 35-copy B1 gene. For the quantitative competitive-PCR, a 410-bp competitor molecule was co-amplified with similar efficiency as the 529bp fragment. Quantitative competitive-PCR produced a linear relationship between the relative amounts of PCR product and the number of tachyzoites in the range of 10(2)-10(4) tachyzoites and 100-3000 tissue cysts. A highly significant correlation between visual counting of brain cysts and quantitative competitive-PCR was obtained in mice chronically infected with Toxoplasma. Thus, quantitative competitive-PCR with the 529bp fragment can be used as an alternative for the tedious visual counting of brain cysts in experimental animals. With the quantitative competitive-PCR, furthermore, we could confirm the copy number of the 529bp fragment in tachyzoites and estimate the number of bradyzoites per cyst. PMID- 10675749 TI - Phylogenetic analysis of the suborder plagiorchiata (Platyhelminthes, Digenea) based on partial lsrDNA sequences. AB - The phylogenetic relationships and systematic position of the members of the suborder Plagiorchiata, one of the derived and most diverse groups of Digenea, have always been controversial. Here, we present a phylogeny of this group based on the analysis of partial sequences of the lsrDNA in 28 species representing 13 families of Plagiorchiata, as well as four outgroups. Our results show that the Plagiorchiata, as considered by most authors, is not monophyletic, and that the superfamilies Opecoeloidea, and most probably Dicrocoelioidea and Gorgoderoidea, may have to be removed from this suborder. According to our results, the Plagiorchiata includes only parasites of terrestrial vertebrates. We find the Plagiorchiata to be composed of two well-supported clades which can be ranked as superfamilies: (1) Plagiorchioidea, including the Plagiorchiidae, Haematoloechidae, Telorchiidae, Brachycoeliidae and Leptophallidae; and (2) Microphalloidea containing the Microphallidae, Prosthogonimidae, Lecithodendriidae and Pleurogenidae. The genetic analysis also allowed revision of the position of several taxa of Plagiorchiata, including: (1) a confirmation of the position of the Brachycoeliidae within the Plagiorchiata; (2) a close phylogenetic relationships of Macrodera with Paralepoderma, Leptophallus and Metaleptophallus; (3) the grouping of Opisthioglyphe and Telorchis within a distinct and strongly supported clade; and (4) the placement of Allassogonoporus amphoraeformis within the Pleurogenidae, and not close to Lecithodendriidae. Some systematic changes, corresponding to these results, are proposed. PMID- 10675748 TI - Molecular cloning and characterisation of a venom allergen AG5-like cDNA from Meloidogyne incognita. AB - RNA fingerprinting was used to identify RNAs that were expressed in parasitic second-stage juveniles of Meloidogyne incognita, but absent from or reduced in preparasitic second-stage juveniles. A cDNA encoding a putative secretory protein was cloned from a M. incognita second-stage juvenile cDNA library by probing with a 0.5kb fragment derived from fingerprinting that was more strongly expressed in parasitic second-stage juveniles. The cDNA, named Mi-msp-1, contained an open reading frame encoding 231 amino acids, with the first 21 amino acids being a putative secretory signal. In Southern blot analysis the Mi-msp-1 hybridised with genomic DNA from M. incognita, Meloidogyne arenaria, Meloidogyne javanica, but not Meloidogyne hapla, Heterodera glycines or Caenorhabditis elegans. In Northern blot analysis a 1kb transcript was detected in both preparasitic and parasitic second-stage juveniles, but not in adult females of M. incognita. Comparing the predicted amino acid sequence with protein databases revealed significant similarity to the venom allergen antigen 5 family of proteins in hymenoptera insects and homologues found in several other nematode species. PMID- 10675750 TI - A molecular systematic framework for equine strongyles based on ribosomal DNA sequence data. AB - In this study, molecular data sets were used to address the controversies relating to the systematics of strongyloid nematodes of equids utilising morphological data sets. DNA sequences of the first and second internal transcribed spacers (ITS-1 and ITS-2) of ribosomal DNA were determined for 30 species of equine strongyles and the systematic relationships reconstructed using phenetic and phylogenetic tree-building methods. The molecular data provided support for the hypothesis that the genera with large subglobular buccal capsules are ancestral to those with small cylindrical buccal capsules, but did not provide support for the current division of the subfamilies Strongylinae and Cyathostominae or for some taxonomic groupings (i.e. generic designations of species) within the Cyathostominea based on morphological data. Although not entirely concordant, the current molecular data provide a systematic framework for future studies of equine strongyles, which could be exploited in combination with new, phylogenetically informative morphological data sets. PMID- 10675751 TI - Inheritance of avermectin resistance in Haemonchus contortus. AB - A larval development assay was used to compare the responses of the Chiswick Avermectin Resistant (CAVRS) isolate of Haemonchus contortus, an avermectin susceptible isolate (VRSG) and their crosses to avermectins. The F(1) and F(2) generations of reciprocal crosses between CAVRS and VRSG were denoted as CAVRS malesxVRSG females=CXV, and VRSG malesxCAVRS females=VXC. The levels of avermectin resistance in the developing larvae of the F(1) of both CXV and VXC were indistinguishable from that in the avermectin-resistant parent, indicating that the resistance trait is completely dominant. Avermectin dose-response curves for the CXV F(1) did not show a 50% mortality rate at low concentrations, indicating that avermectin resistance is not sex-linked. This conclusion was confirmed when adult male worms of the F(1) of the CXV mating were found to have survived treatment of the host with 200microgkg(-1) ivermectin. This dose rate (200microgkg(-1) ivermectin) caused a 50% reduction in the number of adult males in the F(1) from both CXV and VXC crosses, but only a non-significant reduction in the number of adult females in the F(1). Dose-response curves obtained for the F(2) generations in the larval development assay indicated the presence of 25% of avermectin-susceptible individuals, suggesting that a single major gene largely controls the avermectin-resistance trait. This genetic analysis of avermectin resistance in an Australian H. contortus isolate indicates that the expression of the gene for avermectin resistance is an autosomal, complete dominant in the larvae; however, in adults its expression is sex-influenced, with males having a lower resistance to avermectin than females. PMID- 10675752 TI - Enhancement of the human T cell response to culture filtrate fractions of Mycobacterium tuberculosis by microspheres. AB - An improved method of assessment of the human immune response against Mycobacterium tuberculosis antigens will assist the development of new vaccines or diagnostic reagents. In this study, we have analyzed human T cell responses to culture filtrate fractions (CFF) of actively replicating M. tuberculosis strain H37Rv using peripheral blood mononuclear cells from healthy PPD skin test positive and negative individuals. Adsorption of CFF onto polystyrene microspheres, that were approximately the size of the M. tuberculosis (bead adsorbed antigens, BAA) significantly enhanced IFN-gamma production compared to soluble antigens (SA) in PPD skin test positive individuals in an antigen specific manner. Further, BAA induced activation of both CD4(+) and CD8(+) T cell subsets. However, CD4(+) responses in general were higher and their antigenic repertoire was wider than the CD8(+) responses. By contrast, CD8(+) responses were strongest to the lower molecular weight BAA. When CFF were chemically coupled to carboxyl modified microspheres (bead-coupled antigens, BCA), induction of IFN-gamma was similar to BAA. Enhancement of T cell responses to particulate M. tuberculosis antigens may prove useful in vaccine design strategies. PMID- 10675753 TI - Isolation and characterization of human multiple myeloma cell enriched populations. AB - We developed a simple and rapid method to enrich tumor cells within bone marrow (BM) aspirates from patients with multiple myeloma (MM). Thirty patients with a median of 50% (8-85%) MM cells by morphology and 55% (6--85%) MM cells identified by CD38+CD45-cell surface phenotype were studied. BM mononuclear cells (BMMCs) were isolated by Ficoll Hypaque sedimentation and incubated with a cocktail of mouse monoclonal antibodies (mAbs) directed against CD3 (T cells); CD11b and CD14 (monocytes); CD33 (myeloid cells), CD45 and CD45RA (leucocyte common antigen); CD32 as well as glycophorin A. After the addition of anti-mouse Fc Ig-coated immunomagnetic beads, mAb-bound cells were removed in a magnetic field. The residual cell populations were enriched for MM cells, evidenced by >95% plasma cell morphology and >95% CD38+CD45RA-cell surface phenotype. Since this method requires only two short incubations, cell losses were minimal and the yield of MM cells was therefore high (>95%). Viability of the MM-cell enriched fractions was 99%, and these cells were functional in assays of proliferation, cell cycle analysis and immunoglobulin secretion. This immunomagnetic bead depletion method therefore permits the ready isolation of homogeneous populations of patient MM cells for use in both cellular and molecular studies. PMID- 10675754 TI - Evaluation of a high IgE-responder mouse model of allergy to bovine beta lactoglobulin (BLG): development of sandwich immunoassays for total and allergen specific IgE, IgG1 and IgG2a in BLG-sensitized mice. AB - An animal model of food allergy represents an important tool for studying the mechanisms of induction and repression of an allergic reaction, as well as for the development of an immunotherapy to prevent or minimize such an adverse reaction. IgE and IgG1 (Th2 response) vs. IgG2a (Th1 response) are good markers for the induction of an allergic response in mice. Nevertheless, while the total serum concentrations of these isotypes are easy to measure using classical sandwich immunoassays, this is not the case for allergen-specific isotypes. To develop an animal model of allergy to bovine beta-lactoglobulin (BLG), we set up quantitative assays for total and for allergen-specific IgE, IgG1 and IgG2a. Microtiter plates coated either with anti-isotype antibodies (Abs) or with allergen were used for Ab capture, while anti-isotype Fab' fragments coupled to acetylcholinesterase were used for visualization. These assays of anti-BLG specific Abs are original in two ways. First, assay calibration is performed using anti-BLG specific mAbs, thus allowing good quantification of the different isotypes and subclasses of serum antibodies. Second, the detection of all anti BLG specific Abs, i.e., those recognizing both the native and denatured forms of the protein, is achieved through indirect coating of BLG using biotin streptavidin binding. The present assays are quantitative, specific to the isotype (cross-reactivity <0.5%), very sensitive (detection limit in the 10 pg/ml range), and reproducible (coefficient of variation less than 10%). Applied to the humoral response in mice sensitized with BLG adsorbed on alum, these assays proved to be a very useful tool for monitoring high IgE-responder mice following BLG immunization, and for an immunotherapy directed at polarizing the immune response. PMID- 10675755 TI - A critical comparison of frequently used methods for the analysis of tumor necrosis factor-alpha expression by human immune cells. AB - A variety of methods have been developed for the measurement of tumor necrosis factor (TNF)-alpha synthesis by immune cells. Here we have compared the results of the most common used methods, including in vitro stimulation of whole blood or peripheral blood mononuclear cell (PBMC) cultures with phytohaemagglutinin (PHA) or lipopolysaccharide (LPS) and RT-PCR analysis of TNF-alpha transcription in unstimulated PBMC. When we used EDTA treated blood samples we observed a significant correlation between the PHA and LPS stimulated TNF-alpha responses in whole blood or PBMC cultures. In contrast, TNF-alpha concentrations obtained from PHA and LPS stimulated whole blood cultures from citrate-treated blood did not show a correlation. We also found that the PHA stimulated TNF-alpha response was significantly higher in PBMC than in whole blood cultures, whereas the highest LPS stimulated TNF-alpha response was observed in citrate-treated blood. Moreover, the TNF-alpha response in both, citrate and EDTA treated whole blood cultures was significantly higher after LPS than after PHA stimulation. In contrast, in PBMC cultures the PHA stimulated TNF-alpha response was significantly higher than the LPS stimulated response. The results of RT-PCR analysis revealed a significant correlation with the PHA stimulated TNF-alpha response, both in whole blood assays and in PBMC cultures. In addition our results demonstrate that these different methods can only be compared when the influence of external factors such as the immediate processing of blood samples or the use of an appropriate anticoagulant and stimulant is considered. PMID- 10675756 TI - Development and evaluation of a direct sandwich-enzyme-linked immunosorbent assay for the quantification of human hepatic triglyceride lipase mass in human plasma. AB - We have developed a direct sandwich-enzyme-linked immunosorbent assay (ELISA) for quantification of the hepatic triglyceride lipase (HTGL) immunoreactive mass in human plasma. This direct sandwich-ELISA uses a combination of two distinct monoclonal antibodies (MAbs), which recognize different epitopes on the HTGL molecule: a horseradish peroxidase (HRP)-labeled anti-human HTGL MAb (2(4)F12C12) as an enzyme-linked MAb, and an anti-human HTGL MAb (1(11)A3H3) coated on a microtiter plate as a solid-phase MAb. Purified human post-heparin plasma (PHP) HTGL was used as the standard material. The detection range of the sandwich-ELISA was 40-800 ng of HTGL protein per ml of plasma. The intra- and inter-assay coefficients of variation were less than 2.0% and 2.3%, respectively. The recovery tests resulted in variation only between 97.7% and 103.5%. No significant assay interference was caused by a high concentration of triglyceride, hemoglobin, bilirubin, uric acid, or creatinine. The reliability of the HTGL mass values obtained with the direct sandwich-ELISA was assessed by comparison with the HTGL mass values determined by our earlier one-step sandwich enzyme immunoassay (EIA). The two sets of values showed a highly significant correlation (r=+0.952, n=64). Strong correlation (r=+0. 959, n=50) was also found between the HTGL masses with the direct sandwich-ELISA and the HTGL activities determined with a selective immunoinactivation assay. The HTGL mass concentrations in PHP from 64 healthy subjects were 1916+/-841 ng/ml by the direct sandwich-ELISA and 1925+/-785 ng/ml (mean+/-standard deviation (SD)) by the one-step sandwich-EIA. The present direct sandwich-ELISA permits rapid identification of certain HTGL abnormalities in PHP samples from patients with hypertriglyceridemia or diseases such as hypothyroidism or renal failure, which affect HTGL. PMID- 10675757 TI - An ELISA method to measure total and specific human secretory IgA subclasses based on selective degradation by IgA1-protease. AB - We have taken advantage of the property of IgA1-proteases to selectively cleave the human IgA1 subclass into Fabalpha and Fcalpha-J chain-secretory component (Fcalpha-J-SC) fragments in order to design a novel ELISA method for measuring the two secretory IgA (S-IgA) subclasses in secretions. The assay is based on the loss of detection of S-IgA1 by a combination of peroxidase-labelled antibodies to secretory component and Fab following IgA1-protease treatment. The specificity is that of the protease and the sensitivity of the detection is 5 ng/ml. Moreover, the use of purified S-IgA1 and S-IgA2 controls is not necessary. The assay has been successfully applied to the analysis of colostral S-IgA antibodies (Abs) to HIV-1-gp160 from HIV-1 positive women. The major subclass of colostral S-IgA antibodies to gp160 was found to be of the alpha1 isotype but the specific activity of anti-HIV-gp160 S-IgA2 was, however, higher than that of S-IgA1. PMID- 10675758 TI - A simple method to quantify staphylococcal protein A in the presence of human or animal IgG in various samples. AB - Immunoassays designed to measure low concentrations of staphylococcal protein A (SPA) that have been leached into antibody preparations intended for therapeutic use are subject to differing degrees of interference. Methods established to quantify SPA in murine antibody preparations are not accurate in the presence of human or humanized IgG. We report the development of an enzyme-linked immunosorbent assay (ELISA) for SPA with a detection limit of 7 pg/ml and the optimization of a method that permits complete dissociation of SPA-immunoglobulin complexes. This assay is a modification of our heat-mediated dissociation (HD-SD) treatment with sodium dodecyl sulfate (SDS) and diethylenetriaminepentacetic acid (DTPA) for total immune-complex dissociation, in which the heat treatment has been prolonged and the diluent is characterized by increased protein content and buffering capacity. The diluent developed contains SDS, DTPA and bovine serum albumin dissolved in a 0.1 M phosphate buffer (pH 7.2). To validate the efficiency of this novel method, a series of samples have been assayed, including samples reconstituted in vitro, samples of purified antibodies, and plasma from patients. The described method has been shown to be generally efficient in quantitating all native and recombinant SPA in samples containing up to 50 mg/ml of human IgG. These data demonstrate the utility of this technique in determining SPA contamination of recombinant immunoglobulin therapeutic products. PMID- 10675759 TI - The influence of naturally occurring heterophilic anti-immunoglobulin antibodies on direct measurement of serum proteins using sandwich ELISAs. AB - Sandwich ELISAs have become a widely used method for the quantitative detection of serum proteins. However, they can be biased by a variety of interfering substances. As reported recently, we observed false-positive levels of interferon (IFN)-alpha and -beta in up to 27% of sera from healthy blood donors using commercial ELISAs. We now demonstrate that two different groups of naturally occurring heterophilic antibodies (IgG-type) are responsible for these titers. Group I (representing 85% of positive samples) binds to the Fab region of IgG from goat, mouse, rat, horse, and bovidae (but not rabbit). Group II (15%) recognizes an epitope in the Fc region of mouse, horse, bovine, and rabbit (but not goat or rat) immunoglobulins. The antibodies did not crossreact with human IgG subclasses but contributed to false-positive IgG rheumatoid factor levels obtained using a commercially available ELISA. To investigate the susceptibility of assays to these artifacts, various combinations of capture and detection antibodies have been tested. On this basis, we defined the relative risks that standard ELISAs might be influenced by heterophilic anti-immunoglobulin antibodies. In general, assays that use monoclonal antibodies for both capture and detection are less susceptible than others which include at least one polyclonal antiserum. However, only systems utilizing rabbit F(ab')(2) fragments have been found to be immune to this interference. PMID- 10675760 TI - The development of monoclonal human rabies virus-neutralizing antibodies as a substitute for pooled human immune globulin in the prophylactic treatment of rabies virus exposure. AB - To provide a more defined and safer replacement for the human rabies immune globulin (HRIG) from pooled serum which is currently used for treatment of exposure to rabies virus we have developed a series of human rabies virus specific monoclonal antibodies. Mouse-human heterohybrid myeloma cells producing rabies virus-specific human monoclonal antibodies were prepared using B cells obtained from volunteers recently-immunized with a commercial rabies virus vaccine (HDCV). Cell lines producing antibody which neutralized the Evelyn Rokitnicki-Abelseth (ERA) rabies virus strain in vitro were cloned and the resulting monoclonal antibodies characterized for isotype, specificity against a variety of rabies virus isolates, and neutralization capacity. The ability of the monoclonal antibodies to neutralize a variety of rabies virus strains in vitro correlated with their binding specificity for these viruses in an enzyme-linked immunoadsorbant assay (ELISA). A number of these antibodies have proven suitable for the formulation of a prophylactic human monoclonal antibody-based reagent which would provide significant advantages to the HRIG in having defined, reproducible specificity, lessened possibility of contamination with viral pathogens, and consistent availability. PMID- 10675761 TI - Loss of ELISA specificity due to biotinylation of monoclonal antibodies. AB - A significant degree of nonspecificity was found in ELISA determinations of soluble urokinase receptor (suPAR) in human blood plasma when biotinylated monoclonal antibodies (Mabs) were used for the detection layer. Surface plasmon resonance studies using both nonbiotinylated and biotinylated antibodies demonstrated that biotinylation reduced specific binding of the antibodies to their target antigen, suPAR. Furthermore, biotinylation produced a new interaction with unknown human plasma protein(s), unrelated to suPAR. Nonspecific interaction with plasma protein(s) was also observed after biotinylation of a Mab having no specific target antigen in human plasma and, in both cases, the level of nonspecific interaction was directly related to the degree of antibody biotinylation. These results reinforce earlier observations that biotinylation of antibodies can reduce the affinity of antibodies, but also indicate that, in addition, biotinylation can reduce the specificity of immunoassays for plasma proteins. PMID- 10675762 TI - Direct single-step surface plasmon resonance analysis of interactions between small peptides and immobilized monoclonal antibodies. AB - Surface plasmon resonance (SPR) methods have been optimized to permit direct kinetic analysis of the antigenic peptide analytes interacting with immobilized monoclonal antibodies (mAbs). High reproducibility and a significant correlation between SPR and previous ELISA data on the same set of antibodies and peptides were observed. The kinetic data obtained provide further insight into the structure of the main antigenic site of foot-and-mouth disease virus (FMDV). PMID- 10675763 TI - Improved generation of catalytic antibodies by MRL/MPJ-lpr/lpr autoimmune mice. AB - To compare the abilities of different strains of mice to elicit catalytic antibodies (Abs), we determined the occurrence of esterolytic Abs in BALB/c (normal strain) and MRL/MPJ-lpr/lpr (MRL/lpr, autoimmune) mice after immunization with the transition state analog (TSA) 1. Hybridoma supernatants elicited against TSA 1 were screened by ELISA for binding to the BSA-conjugated TSA 1 (=3b), and then screened for binding to the BSA-linked short TSA 2 (=4). We obtained eight times more positives from MRL/lpr mice than from BALB/c mice by these screening steps. The monoclonal antibodies (mAbs) obtained here were examined for binding and catalytic activity. Fifteen of 25 mAbs from MRL/lpr had esterolytic activity, compared with only two of 21 mAbs from BALB/c. These results demonstrated that the occurrence of catalytic Abs was much higher in MRL/lpr mice than in BALB/c mice, which is in good agreement with the previous report by Tawfik et al. [Tawfik, D.S., Chap, R., Green, B.S., Sela, M., Eshhar, Z., 1995. Proc. Natl. Acad. Sci. U.S.A. 92, 2145-2149] using a different kind of TSA. Thus, these studies strongly suggest that using the appropriate strain can be a key factor in the efficient production of catalytic Abs. Furthermore, these mAbs were characterized to elucidate the mechanism of strain difference, and determine whether MRL/lpr mice can be used with other TSAs for the efficient production of catalytic Abs. PMID- 10675764 TI - Measurement of lymphocyte subset proliferation by three-color immunofluorescence and DNA flow cytometry. AB - We developed a method for simultaneous flow cytometric analysis of three-color immunofluorescence and DNA content. We show here that staining with 7-amino actinomycin D (7-AAD) at 10 microg/ml using a phosphate-citrate buffer at low pH containing saponin for cell membrane permeabilization yields good resolution DNA histograms with low coefficients of variation. Furthermore, light scatter properties of cells are preserved after permeabilization; this permits gating on cell populations that differ in scatter signals on the flow cytometer. Because of the low pH of the phosphate-citrate staining buffer, Alexa488, a pH-independent green-fluorescent fluorochrome is used instead of fluorescein-isothiocyanate (FITC) for cell surface staining in combination with phycoerythrin (PE) and with allophycocyanin (APC) which are both pH insensitive. Removal of 7-AAD after staining and replacing it with non-fluorescent actinomycin D (AD) retains DNA staining and allows detection of Alexa488, PE and APC cell surface immunofluorescence without interference from fluorescent 7-AAD in solution for clear identification of cell subpopulations even after prolonged stimulation in culture. Thus, using a four-color benchtop flow cytometer, measurement of Alexa488, PE and APC three-color immunofluorescence can be combined with 7-AAD DNA content analysis. Furthermore, we demonstrate that sample storage overnight without fixation for later analysis on the flow cytometer is possible without compromising results. Application of the method to the assessment of the differential proliferative responses of lymphocyte subsets of human peripheral blood mononuclear cells (PBMC) that were costimulated with CD3 and with CD28.2 is presented. PMID- 10675765 TI - Differential expression of S100B and S100A6(1) in the human fetal and aged cerebral cortex. AB - S100B and S100A6 (calcylin) are two members of the S100 Ca(2+)-binding protein family and have been localized in the mammalian nervous system. However, information on their distribution in the human nervous system, especially in the developing human fetal brain, is scarce. In the present study, an immunocytochemical method was used to examine the spatio-temporal protein expression patterns of S100B and S100A6 in normal human fetal hippocampus, entorhinal cortex and occipital cortex. Normal aged adult human brain specimens were also included for comparison. From week 15 onwards, an increase with advancing gestation age in both the number and staining intensity of S100B positive, astrocyte-like cells was found in the pyramidal layer of the hippocampus, while both the molecular and polymorphic layers showed similar S100B immunoreactivities at all stages examined. A decrease in the immunoreactivities was found in the molecular layer of the aged adult hippocampus while other layers exhibited immunoreactivities similar to those of the late fetus. At week 15, the molecular, pyramidal and ganglionic/multiform layers of the entorhinal cortex also showed positive S100B immunoreactivities which were maintained throughout the rest of the gestation and in adult specimens. In the occipital cortex, the numbers of positive cells for all layers were about twofold higher than those found in the hippocampus and entorhinal cortex, and immunoreactivities detected in the granular layer increased from week 21, reaching a plateau at around week 27. S100B positive fibers were also found at week 30 but were not observed in aged adult specimens. S100A6 positive cells were on the whole fewer in number than those of S100B in the brain regions examined. The S100A6 immunoreactivities which were localized in some pyramidal neuron-like and some glial-like cells of the pyramidal and molecular layers of the hippocampus increased by midgestation and became weak in the late fetus and in aged adult specimens. Weakly stained S100A6 positive cells were also observed in the entorhinal cortex throughout the gestation and in aged adult cortex. S100A6 immunoreactivities were weak in the fetal occipital cortex. They were also localized in the glial-like cells of the aged adult occipital cortex. The differential spatio-temporal expression of S100B and S100A6 proteins suggests that the proteins play different roles in different brain regions during development and in adulthood. PMID- 10675766 TI - Effects of hindlimb unloading on neuromuscular development of neonatal rats. AB - We hypothesized that hindlimb suspension unloading of 8-day-old neonatal rats would disrupt the normal development of muscle fiber types and the motor innervation of the antigravity (weightbearing) soleus muscles but not extensor digitorum longus (EDL) muscles. Five rats were suspended 4.5 h and returned 1.5 h to the dam for nursing on a 24 h cycle for 9 days. To control for isolation from the dam, the remaining five littermates were removed on the same schedule but not suspended. Another litter of 10 rats housed in the same room provided a vivarium control. Fibers were typed by myofibrillar ATPase histochemistry and immunostaining for embryonic, slow, fast IIA and fast IIB isomyosins. The percentage of multiple innervation and the complexity of singly-innervated motor terminal endings were assessed in silver/cholinesterase stained sections. Unique to the soleus, unloading accelerated production of fast IIA myosin, delayed expression of slow myosin and retarded increases in standardized muscle weight and fiber size. Loss of multiple innervation was not delayed. However, fewer than normal motor nerve endings achieved complexity. Suspended rats continued unloaded hindlimb movements. These findings suggest that motor neurons resolve multiple innervation through nerve impulse activity, whereas the postsynaptic element (muscle fiber) controls endplate size, which regulates motor terminal arborization. Unexpectedly, in the EDL of unloaded rats, transition from embryonic to fast myosin expression was retarded. Suspension-related foot drop, which stretches and chronically loads EDL, may have prevented fast fiber differentiation. These results demonstrate that neuromuscular development of both weightbearing and non-weightbearing muscles in rats is dependent upon and modulated by hindlimb loading. PMID- 10675767 TI - Effects of neonatal rat Borna disease virus (BDV) infection on the postnatal development of the brain monoaminergic systems. AB - Effects of neonatal Borna disease virus infection (BDV) on the postnatal development of brain monoaminergic systems in rats were studied. Tissue content of norepinephrine (NE), dopamine (DA) and its metabolite, 3,4-dihydroxyphenol acetic acid (DOPAC), and serotonin (5-HT) and its metabolite, 5-hydroxyindole-3 acetic acid (5-HIAA) were assayed by means of HPLC-EC in frontal cortex, cerebellum, hippocampus, hypothalamus and striatum of neonatally BDV-infected and sham-inoculated male Lewis rats of 8, 14, 21, 60 and 90 days of age. Both NE and 5-HT concentrations were significantly affected by neonatal BDV infection. The cortical and cerebellar levels of NE and 5-HT were significantly greater in BDV infected rats than control animals at postnatal days (PND) 60 and 90. Tissue content of NE in hippocampus was unaffected. In hippocampus, neonatally BDV infected rats had lower 5-HT levels at PND 8 and significantly elevated levels at PND 21 and onwards. Neither striatal levels of 5-HT nor hypothalamic levels of 5 HT and NE were affected by neonatal BDV infection, suggesting that the monoamine systems in the prenatally maturing brain regions are less sensitive to effects of neonatal viral infection. 5-HIAA/5-HT ratio was not altered in BDV-infected rats indicating no changes in the 5-HT turnover in the brain regions damaged by the virus. Neither DA nor DOPAC/DA ratio was affected by neonatal BDV infection in any of the brain regions examined. The present data demonstrate significant and specific alterations in monoaminergic systems in neonatally BDV-infected rats. This pattern of changes is consistent with the previously reported behavioral abnormalities resulting from neonatal BDV infection. PMID- 10675768 TI - Gene expressions during the development and sexual differentiation of the olfactory bulb in rats. AB - In this study, expressions of cell-cycle-related genes: p53, retinoblastoma (Rb), p21, bcl-2(alpha), bcl-2(beta); protooncogene c-ski; glial cell marker protein gene S100beta; neurotransmitter gene, substance P and sexual-differentiation related genes, androgen receptor (AR) and estrogen receptor beta (ER(beta)), are studied in the olfactory bulb of groups of both six female and six male rats at the ages of 3, 10, 20 and 40 days. Expressions of housekeeping genes such as beta actin, cyclophilin and proliferating cell nuclear antigens (PCNA) are determined using reverse transcription polymerase chain reaction (RT-PCR) for the correction of unequal amount of cDNA added into the samples. Using labeled 32P-dCTP and Phosphorimager technology, relative abundance of radioactivities of the PCR products is obtained by dividing the radioactivity of each individual sample by the corresponding radioactivities of different housekeeping genes. Data evaluated by Two-way ANOVA indicate that only the bcl-2(alpha) gene expression is affected significantly by age, sex and their interactions no matter which of the three housekeeping genes is used for correction. When beta-actin was used for corrections, effects of age but not sex were found in the expressions of p53, Rb, p21, AR, ER(beta), substance P and S100beta genes, but not in bcl-2(beta), c-ski, cyclophilin and PCNA genes. While cyclophilin was used for corrections, only the p53, Rb, AR, ER(beta), substance P and S100beta but not the bcl-2(beta), p21, c ski, PCNA and beta-actin genes are affected by age. They are all not influenced by sex of the animals. Only the AR, ER(beta) and S100beta genes are age-dependent when PCNA was used for the correction. The other gene expressions are not altered by sex, while the interactions of age and sex were found to be significantly affecting the bcl-2(beta) gene expression. Conclusively, developmental changes of the p53, Rb, AR, ER(beta), substance P and S100beta genes expressions are quite evidenced while only the bcl-2(alpha) gene seems to change significantly during the sexual differentiation of olfactory bulb in rats. PMID- 10675769 TI - Comparative immunocytochemical study of FMRFamide neuronal system in the brain of Danio rerio and Acipenser ruthenus during development. AB - The distribution of FMRFamide-like immunoreactive (ir) neurons and fibers was investigated in the central nervous system of developing zebrafish and juvenile sturgeon (sterlet). Adult zebrafish was also studied. In zebrafish embryos FMRFamide-ir elements first appeared 30 h post-fertilization (PF). Ir somata were located in the olfactory placode and in the ventral diencephalon. FMRFamide-ir fibers originating from diencephalic neurons were found in the ventral telencephalon and in ventral portions of the brainstem. At 48 h PF, the ir perikarya in the olfactory placode displayed increased immunoreactivity and stained fibers emerged from the somata. At 60 h PF, bilaterally, clusters of FMRFamide-ir neurons were found along the rostro-caudal axis of the brain, from the olfactory placode to rostral regions of the ventro-lateral telencephalon. At 60 h PF, numerous ir fibers appeared in the dorsal telencephalon, optic lobes, optic nerves, and retina. Except for ir fibers in the hypophysis at the age of 72 h PF, and a few ir cells in the nucleus olfacto-retinalis (NOR) at the age of 2 months PF, no major re-organization was noted in subsequent ontogenetic stages. The number of stained NOR neurons increased markedly in sexually mature zebrafish. In adult zebrafish, other ir neurons were located in the dorsal zones of the periventricular hypothalamus and in components of the nervus terminalis. We are inclined to believe that neurons expressing FMRFamide originate in the olfactory placode and in the ventricular ependyma in the hypothalamus. On the same grounds, a dual origin of FMRFamide-ir neurons is inferred in the sturgeon, an ancestral bony fish: prior to the observation of ir cells in the nasal area and in the telencephalon stained neurons were noted in circumventricular hypothalamic regions. PMID- 10675770 TI - Glial-derived neurotrophic factor (GDNF) prevents ethanol-induced apoptosis and JUN kinase phosphorylation. AB - Ethanol exposure during neural development leads to substantial neuronal loss in multiple brain regions. Our previous research indicated that exogenous glial derived neurotrophic factor (GDNF) attenuated ethanol-induced cerebellar Purkinje cell loss. Additionally, ethanol decreased GDNF release suggesting that ethanol disrupts GDNF-signaling pathways. The present experiments utilized a homogeneous GDNF-responsive neuroblastoma cell line (SK-N-SH) to test the hypothesis that exogenous GDNF could attenuate ethanol-induced cell loss by suppressing cytotoxic signaling pathways and cell suicide. We measured two independently regulated markers of apoptosis, DNA fragmentation and the externalization of phosphatidylserine to the outer cell membrane leaflet. Ethanol induced a dose related increase in both apoptosis and necrosis. Lower concentrations of ethanol (34 and 68 mM) specifically increased DNA fragmentation, while all concentrations (up to 137 mM) increased phosphatidylserine translocation, suggesting that ethanol induction of apoptosis is not a unitary process. Furthermore, only higher concentrations of ethanol (103 and 137 mM) induced necrosis. Additionally, ethanol specifically induced phosphorylation of c-jun N-terminal-kinase (JNK), a mitogen-activated protein (MAP) kinase selectively associated with apoptosis. In contrast, ethanol did not alter the phosphorylation of another MAP kinase, the extracellular signal-regulated kinases (ERK) that mediate cell survival. Thus, ethanol activated specific intracellular cell death-associated pathways and induced cell death. GDNF, in turn, prevented both ethanol-induced apoptosis and the activation of the death-associated JNK cascade. Therefore, GDNF may regulate multiple pathways to prevent ethanol-induced cell loss. PMID- 10675771 TI - Developmental expression of urinary bladder neurotrophic factor mRNA and protein in the neonatal rat. AB - These studies were performed to determine the developmental expression pattern of neurotrophic factor (NTF: nerve growth factor (betaNGF), brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF), ciliary neurotrophic factor (CNTF), neurotrophin-3 (NT-3) and NT-4 mRNA and NGF, NT-3 and NT-4 protein in the urinary bladder of the postnatal Wistar rat. It was hypothesized that NTFs may contribute to the development of the spinobulbospinal micturition reflex that represents the adult micturition pattern. Changes in NTF mRNA or protein expression in the urinary bladder at the time of development of the mature micturition reflex (postnatal days (P) 16-18) may suggest an involvement of target-derived NTFs in this maturation process. Developmental ages, prior to (P5, P10, P15) or following (P20, P30, adult P90) the development of the spinobulbospinal micturition reflex were selected and the urinary bladder was analyzed for levels of neurotrophic factor mRNA or protein. Results from ribonuclease protection assays demonstrated a similar developmental pattern among each neurotrophic factor examined. Neurotrophic factor mRNA levels increased by P10 and reach a maximum by P15. Subsequently, NTF mRNA levels declined to adult levels that were less than the earliest postnatal time examined (P5). NTF mRNA expression was significantly (p50%. In contrast, intranigral application of picrotoxin, a GABA antagonist, had the opposite effect. Neither GABA nor picrotoxin altered striatal 3,4-dihydroxyphenylacetic acid (DOPAC), a major dopamine metabolite. Collectively, these results indicate that intranigral GABA exerts a tonic inhibitory influence on ascorbate release in the striatum. PMID- 10675794 TI - Laterality effects in selective attention to threat after repetitive transcranial magnetic stimulation at the prefrontal cortex in female subjects. AB - Recently, several experiments have indicated that the left and right prefrontal cortex (PFC) are differently involved in emotional processing. The aim of this study was to investigate the role of the left and right PFC in selective attention to angry faces by using a pictorial emotional Stroop task. Slow repetitive transcranial magnetic stimulation (rTMS) was applied to the left and right PFC of 10 female subjects for 15 min on separate days. Results showed a significant effect of stimulation position: right PFC rTMS resulted in selective attention towards angry faces, whereas left PFC rTMS resulted in selective attention away from angry faces. This finding is in accordance with theoretical accounts of the neural implementation of approach and withdrawal systems. PMID- 10675795 TI - Diurnal preference, sleep habits, circadian sleep propensity and melatonin rhythm in healthy human subjects. AB - After 24-h sleep deprivation, 33 healthy young subjects entered the 10/20 min ultra-short sleep-wake schedule for 26 h. Melatonin rhythm was hourly assessed simultaneously. Results indicated that morning preference was significantly correlated with habitual sleep onset (r=-0.41, P=0.04), habitual sleep offset (r= 0.52, P=0.002), melatonin peak time (r=-0.36, P=0.04), and sleep propensity onset time (r=-0.36, P=0.04). The intervals between habitual sleep mid-point and melatonin peak time and between habitual sleep mid-point and sleep propensity onset time were significantly longer in morning-preference subjects than in evening-preference subjects (P<0.05). These findings suggest that the variance of diurnal preference may be related to differences in phase relations between habitual sleep timing and the circadian pacemaker. PMID- 10675796 TI - Inhibition by neuropeptide Y of fentanyl-induced muscular rigidity at the locus coeruleus in rats. AB - The aim of this study was to evaluate the effects of centrally administered neuropeptide Y (NPY) on muscular rigidity induced by fentanyl in Sprague-Dawley rats. They were anesthetized with ketamine (120 mg/kg, i.p.) and their lungs were mechanically ventilated. Intravenous administration of fentanyl (100 microg/kg) consistently evoked a significant increase in the electromyographic activity (EMG) recorded from the sacrococcygeus dorsalis lateralis muscle. This implied muscular rigidity was appreciably attenuated by intracerebroventricular administration of NPY (4 nmol/2.5 microl). Microinjection of NPY (40 or 160 pmol/50 nl) into the bilateral locus coeruleus (LC) elicited an inhibition of the EMG activation induced by fentanyl in a dose-dependent manner. Microinjection of 160 pmol NPY plus antiserum against NPY (NPY(Ab), 1:20) into the LC failed to suppress fentanyl-induced muscular rigidity. However, this implied muscular rigidity was not affected by NPY plus normal rabbit serum (NRS, 1:20). In addition, NPY(Ab) or NRS per se had no substantial effect on the rigidity. Microinjection of NPY (160 pmol) into the areas adjacent to the LC did not attenuate the rigidity. Our results suggest that the centrally administered NPY attenuated fentanyl-induced muscular rigidity by acting at the LC, but endogenous NPY may not be involved in this process. PMID- 10675797 TI - Polysialylated neural cell adhesion molecule modulates photic signaling in the mouse suprachiasmatic nucleus. AB - Polysialic acid (PSA), a sialic acid polymer that regulates plasticity and cell cell interactions in neural tissues, is expressed in the mammalian circadian clock located in the suprachiasmatic nucleus (SCN). In vivo enzymatic removal of PSA from the mouse SCN significantly impaired both the photic induction of Fos protein in SCN cells and light-induced phase-resetting of the circadian locomotor activity rhythm. Genetic deletion of PSA and it's neural cell adhesion molecule (NCAM) carrier correspondingly attenuated light-induced circadian phase-shifting. Comparison of PSA levels between young and old mice revealed a large aging related reduction in SCN PSA content that accompanies the diminished capacity for circadian photic response reported in old rodents. Collectively these data support the contention that PSA modulates photic signaling in the SCN, and that normal reductions in the cell surface molecule contribute to aging-related deficits in SCN circadian clock function. PMID- 10675798 TI - Repeated administration of systemic gabapentin alleviates allodynia-like behaviors in spinally injured rats. AB - The effect of systemic gabapentin, a novel antiepileptic and analgesic, was tested on allodynia-like behaviors in spinal cord injured rats. On the first day of treatment 30 mg/kg intraperitoneal gabapentin did not alleviate hyper reactivity to mechanical and cold stimulation. The allodynia was significantly reduced by 100 mg/kg gabapentin, which however, produced sedation and motor impairments. Repeated administration of 30 mg/kg gabapentin once a day produced a gradually increasing anti-allodynic effect. Total alleviation of mechanical allodynia was observed in most rats after the third administration of gabapentin. Thus, build-up of the antiallodynic effect of gabapentin may develop through a time dependent mechanism or alternatively through a gradual accumulation of the effective central nervous system concentration of the drug. PMID- 10675799 TI - Gender-specific association of the angiotensin converting enzyme gene with Alzheimer's disease. AB - Epidemiological studies have demonstrated that risk factors for vascular disease are also risk factors for Alzheimer's disease (AD). The gene for the angiotensin converting enzyme (ACE) has recently been reported to be associated with risk for AD. We have investigated the possibility of such an association in 98 clinic based and 73 community-based AD cases versus 175 community-based controls and find a gender-specific association of ACE genotype with AD in the female clinic population. These data suggest that gender may interact with genetic factors to influence risk for AD. Gender-specific risk for AD has been previously reported, and a biological rationale for involvement of ACE in the AD process is supported by studies exploring the relationship between AD and vascular risk factors such as hypertension. However, the results may also be a consequence of the known anomalies that arise in genetic association studies as a consequence of sample selection. PMID- 10675800 TI - Differential neurotrophin levels in cerebrospinal fluid and their changes during development in newborn rat. AB - Cerebrospinal fluid concentration of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) was measured in normal developing rat from birth to postnatal day (PND) 21 by enzyme-linked immunosorbent assay. NGF levels were significantly higher than those of BDNF and NT-3 from PND 1-21. NGF levels decreased from PND 1-3 to PND 9. At PND 15 and 17, NGF levels peaked a second time and rapidly decreased to PND 21. BDNF peaked at PND 13-15, while NT-3 levels peaked at PND 7-9. Each of the three neurotrophins has its own characteristic pattern in changes in cerebrospinal fluid levels. PMID- 10675801 TI - Cloning of serotonin 5-HT(1) receptor subtypes from the chimpanzee, gorilla and Rhesus monkey and their agonist-induced guanosine 5'gamma(35)S triphosphate binding. AB - 5-HT(1) receptor subtypes ((1B), (1D) and (1F)) have been implicated in migraine pathophysiology and their ligands have been examined for pharmacological actions in various experimental animal models. Considerable divergences exist, however, in their primary sequences between experimental animals and human, and additional models closer to human, such as non-human primates seem to be useful for migraine research. Earlier, we cloned the 5-HT(1D), and here 5-HT(1B) and 5-HT(1F) receptors from the chimpanzee, gorilla and Rhesus monkey, via polymerase chain reactions with their genomic DNAs and primers designed from the corresponding human receptors. Direct sequencing of PCR products showed that the 5-HT(1B) receptors from the chimpanzee, gorilla and monkey differ from the human receptor by 0, 1 and 7 residues, respectively while 5-HT(1F) receptors differ by 0, 3 and 10 residues, respectively. These divergent residues are mostly conservatively substituted and also largely confined to the N-terminal region and the 3rd intracellular loop, away from transmembrane segments and intracellular loops near membrane which are critical for ligand binding and G protein coupling. The chimpanzee 5-HT(1D), 5-HT(1B) and monkey 5-HT(1F) receptors, as heterologously expressed in human embryonic kidney 293 cells, showed robust agonist-induced guanosine 5'gamma (35)S triphosphate (GTPgamma(35)S) binding through activation of G proteins containing Ggamma(i) subunits. Moreover, pronounced inhibition of basal GTPgamma(35)S binding by methiothepin (an antagonist), representing constitutively active receptors, was observed with only 5-HT(1D). Overall, ligand binding and GTPgamma(35)S binding profiles for these primate receptors are comparable to those for the human receptors and validate non-human primates as useful models for human migraine research. PMID- 10675807 TI - Coordination of two- and one-joint muscles: functional consequences and implications for motor control. AB - The purpose of this paper is three-fold: (a) to summarize available data on coordination of major two- and one-joint muscles in multijoint tasks and identify basic features of muscle coordination, (b) to demonstrate that there may exist an optimization criterion that predicts essential features of electromyographic activity of individual muscles in a variety of tasks, and (c) to address the functional consequences of the observed muscle coordination and underlying mechanisms of its control. The analysis of the literature revealed that basic features of muscle coordination are similar among different voluntary motor tasks and reflex responses. It is demonstrated that these basic features of coordination of one- and two-joint muscles in two-dimensional tasks are qualitatively predicted by minimizing the sum of muscle stresses cubed. Functional consequences of the observed coordination of one- and two-joint muscles are (a) reduction of muscle force as well as stress, mechanical and metabolic energy expenditure, muscle fatigue, and perceived effort; (b) a spring like behavior of a multi-joint limb during maintenance of an equilibrium posture; and (c) energy transfer between joints via two-joint muscles. A conceptual scheme of connections between motoneuron pools of one- and two-joint muscles, which accounts for the observed muscle coordination, is proposed. An important part of this scheme is the force-dependent inhibition and excitation from two-joint to one-joint synergists and antagonists, respectively. PMID- 10675808 TI - Energy-minimizing choices of muscles and patterns of movement. AB - Prilutsky (1999, target paper) reports that Crowninshield and Brand's (1981) criterion, minimization of the sum of the cubes of muscle stresses, works well as a predictor of the division of labor between muscles, for various tasks. However, no direct benefit from minimizing this particular sum is apparent, and it seems likely that it is merely a correlate of the criterion that actually drives muscle choice. In many tasks, there would be a clear, direct benefit from minimizing metabolic energy costs, as Prilutsky (1999) points out. Alexander (1997a, 1997b) and Minetti and Alexander (1997) have shown how the metabolic energy costs of muscle contraction can be estimated, and used to predict optimum muscle properties or optimal patterns of movement. This article explores the feasibility of using the same approach to predict optimum division of labor between one- and two-joint muscles. PMID- 10675809 TI - Two-joint muscles offer the solution, but what was the problem? AB - Prilutsky's paper is mainly concerned with the coordination of one- and two-joint muscles. This commentary on the paper addresses the question why we have two joint muscles in the first place. From an evolutionary point of view, two-joint muscles must have contributed to fitness by presenting a solution to problems that could not be solved with musculoskeletal systems comprising only one-joint muscles. One such problem, not mentioned by Prilutsky, is the following. In a system equipped with only one-joint muscles, satisfying directional constraints would demand, in certain phases of movements, deactivation of muscles that are shortening. Consequently, the work output of these muscles would be limited. The incorporation of two-joint muscles helps to overcome this problem. The reason is that it offers the possibility to redistribute energy across joints, thereby making it possible to accomplish more successfully the difficult task of producing work while steering the movement. PMID- 10675810 TI - Too early to explain all experimental data with a single model. AB - The target article by Prilutsky gives an excellent overview of the predictions of the Crowninshield and Brand model and about the relevant literature about muscle coordination. However, we do not agree with the claim that the Crowninshield and Brand model can explain the coordination between one-joint and two-joint muscles. In this commentary we will make three claims: (a) The Crowninshield and Brand model cannot explain all aspects of muscle coordination, (b) there is good experimental evidence that different constraints and models may be necessary to explain muscle coordination in different motor tasks, and (c) the reason for the lack of quantitative fits between predictions about muscle force and experimental data is that it is hard to measure muscle force in man. As a compromise one has to rely on EMG activity as a measure of muscle force. Because of the complex relationship between EMG and muscle force, a quantitative test of models is difficult. PMID- 10675811 TI - Minimizing stress is not enough. AB - Muscle stress is plainly one of the physical variables that the central nervous system probably wishes to minimize. This criterion does not uniquely define the patterns of muscle activation. It fails to explain the degree of coactivation of muscle antagonists that is widely found, and it cannot explain why two movements or movement segments that follow an identical trajectory driven by identical joint torques can be driven by different patterns of muscle activation. Muscle contraction provides for both net joint torque and limb stability. The minimization of the sum of muscle stresses, raised to any power, is an insufficient rule. PMID- 10675812 TI - Coordination of one- and two-joint muscles during voluntary movement: theoretical and experimental considerations. AB - The target article by Dr. Prilutsky is based on three incorrectly derived mathematical rules concerning force-sharing among synergistic muscles associated with a cost function that minimizes the sum of the cubed muscle stresses. Since these derived rules govern all aspects of Dr. Prilutsky's discussion and conclusion and form the basis for his proposed theory of coordination between one and two-joint muscles, most of what is said in the target article is confusing or misleading at best or factually wrong at worst. The aim of our commentary is to sort right from wrong in Dr. Prilutsky's article within space limitations that do not allow for detailed descriptions of mathematical proofs and explicit discussions of the relevant experimental literature. PMID- 10675813 TI - General coordination principles elucidated by forward dynamics: minimum fatique does not explain muscle excitation in dynamic tasks. AB - The target article presents a framework for coordination of one- and two-joint muscles in a variety of tasks. Static optimization analyses were performed that minimize muscle fatigue, and it is claimed that the predicted muscle forces account for essential features of EMG activity qualitatively well. However, static optimization analyses use the observed joint moments, which implicitly assumes that they minimize the total muscle fatigue of the task. We use a forward dynamics (i.e., relationship between muscle forces and the kinematics and kinetics of task performance) modeling approach to show that this assumption does not appear to be true in cycling (which was used as an example task in the target article). Our results challenge the hypothesized coordination framework and the underlying concept that general coordination principles for dynamic tasks can be elucidated using inverse-dynamics-based analyses. PMID- 10675814 TI - Overcomplete musculature or underspecified tasks? AB - The number of muscles in the body is actually fairly close to the number required to control completely all its degrees of freedom. The apparent need for a coordinating principle arises from the experimental practice of asking subjects to perform simple movements and assuming that they make no implicit assumptions about other constraints. Natural activities include implicit constraints that differ greatly for different tasks and circumstances and that would be met best by a nervous system free of a priori principles. PMID- 10675815 TI - Subject specific coordination of two- and one-joint muscles during landings suggests multiple control criteria. AB - The target article, thoughtfully constructed by Dr. Prilutsky, effectively synthesizes available data on multijoint movements regarding coordination patterns of major two- and one-joint muscles, provides evidence for an optimization criterion that predicts critical features of muscle activation patterns, and explores the functional consequences of muscle coordination. This work also provides a clear set of definitions and an organizational framework that is currently needed for a productive interdisciplinary discussion regarding the underlying control mechanisms used during realistic multijoint movements. Although identification of an optimization criterion that predicts muscle recruitment strategies would greatly simplify control logic required for rehabilitation and musculoskeletal modeling, our experimental data during landings indicate more than one criterion may exist. Preliminary review of our experimental landing data suggests the rules identified by Prilutsky apparently hold for some subjects during portions of the landing movements. The presence of more than one muscle activation pattern used to achieve the same NJMs demonstrates there may be more than one optimization criterion that predicts critical features of muscle activation patterns. The functional consequences of more than one control criterion may also prove to be an asset, particularly when adapting to different environmental constraints. PMID- 10675816 TI - Some mechanical considerations on muscle coordination. AB - This commentary emphasizes three points of discussion. (a) The terminology: The terms multifunctional, synergisic, antagonistic muscles, and synergistic and antagonistic coactivations are discussed and the conclusion is drawn that they could not be used without mentioning the particular joint motion. (b) The importance of the external joint moments for activation of the muscles is confirmed on the basis of logical and mechanical considerations. Not all experimental results, however, could be explained by this means. (c) The optimization criterion: Prilutsky's conclusion concerning the predicted muscle force proportionality to the muscle moment arm and PCSA is confirmed using a simple analytical solution of the optimization problem. It is shown, however, that the proportionality to the PCSA is a consequence of the chosen optimization criterion. PMID- 10675817 TI - Muscle coordination: the discussion continues AB - In this response, the major criticisms of the target article are addressed. Terminology from the target article that may have caused some confusion is clarified. In particular, the tasks that have the basic features of muscle coordination, as identified in the target article, have been limited in scope. A new metabolic optimization criterion suggested by Alexander (2000) is examined for its ability to predict muscle coordination in walking. Issues concerning the validation of muscle force predictions, the rules of muscle coordination, and the role of directional constraints in coordination of two-joint muscles are discussed. It is shown in particular that even in one-joint systems, the forces predicted by the criterion of Crowninshield and Brand (1981) depend upon the muscle moment arms and the physiological cross-sectional areas in much more complex ways than either previously assumed in the target article, or incorrectly derived by Herzog and Ait-Haddou (2000). It is concluded that the criterion of Crowninshield and Brand qualitatively predicts the basic coordination features of the major one- and two-joint muscles in a number of highly skilled, repetitive motor tasks performed by humans under predictable conditions and little demands on stability and accuracy. A possible functional significance of such muscle coordination may be the minimization of perceived effort, muscle fatigue, and/or energy expenditure. PMID- 10675818 TI - [How can we limit the number of unnecessary appendectomies?]. PMID- 10675819 TI - [Management of Barrett's esophagus]. AB - The subject of this review concerns advances in the diagnosis, etiology, complications, treatment and surveillance of Barrett's esophagus. Current management of Barrett's esophagus is discussed and indications of surgical treatment are clarified while specific destruction of Barrett's mucosa is possible by endoscopic management (laser or photodynamic therapy). PMID- 10675820 TI - [Chemotherapy, combined radiochemotherapy and new therapeutic approaches in adenocarcinoma of the pancreas]. AB - For patients having undergone complete resection for adenocarcinoma of the pancreas, combined radiochemotherapy protocols using bolus 5FU as neoadjuvant or adjuvant treatments can help control disease spread and perhaps moderately lengthen survival. As the rare controlled trials having tested these therapeutic strategies have provided conflicting data, this therapeutic attitude cannot be considered as a standard treatment. The tested protocols using combined radiochemotherapy were developed in the sixties and seventies and have been greatly improved since that time. New combinations for neoadjuvant and adjuvant radiochemotherapy protocols are currently under evaluation in controlled therapeutic trials. Systemic chemotherapy (gemcitabine, 5FU, platinum) has a palliative effect, improving the quality of life in patients with advanced-stage disease. Gemcitabine is easy to administer and has a low toxicity profile. It is widely used in standard protocols. Therapeutic trials combining gemcitabine and other cytotoxic agents are under way. Radiochemotherapy combinations using 5FU are a palliative alternative for patients with locally advanced disease, particularly those with painful symptoms. There is an urgent need for more effective treatments against metastatic disease and for better loco-regional management using a multidisciplinary approach. These patients should be treated within the framework of therapeutic trials. PMID- 10675821 TI - [Surgical techniques. Laparoscopic right colectomy]. PMID- 10675822 TI - [Surgical techniques. Left pancreatectomies]. PMID- 10675823 TI - [Images in surgery. Benign florid corticoadrenaloma]. PMID- 10675824 TI - [The world's first: laparoscopic portacaval H-graft shunt]. AB - Partial portal diversion prevents recurrent variceal hemorrhage in cirrhotic patients. Surgical portacaval shunt and trans-jugular intra-hepatic portasystemic shunt have both been realized. Endoscopic instrument's advances and surgeon's skill level in laparoscopic surgery had overcome technical impediments. Laparoscopy could be used in safety condition in shunt procedures. We present the case report of surgical laparoscopic procedure for portacaval H graft shunt. PMID- 10675825 TI - [Pseudomyxoma peritonei. A review]. AB - Pseudomyxoma peritonei is a rare disease characterized by the presence of a large mucin component within the abdomen. Recent pathological and genetic advances indicate that they originate from an appendiceal adenoma or adenocarcinoma. Their prognostic is worse than ovarian border-line mucinous tumors (with which they are frequently confused). Currently, the histologic aspect permits to separate the diffuse peritoneal adenomucinosis (DPAM) originating from adenomas, with a relatively benign course, from the mucinous peritoneal carcinomatosis (MPC) originating from adenocarcinomas, with a very poor prognosis. Paradoxically, the treatment of these two diseases are rather similar, with supra-radical surgery as frequently as possible. This type of surgery allows to reach a crude 5-year survival comprised between 50% and 70%, with very different results according to the DPAM-type or the MPC-type. The adjunction of an intraperitoneal chemo hyperthermia is logical for these two types of disease and probably increases the survival rate. PMID- 10675827 TI - [In Process Citation] PMID- 10675826 TI - [Congress of the American College of Surgeons (ACS). San Francisco, 10-15 October 1999]. PMID- 10675828 TI - [Initial access for laparoscopic gynecologic surgery. French Society of Endoscopic Gynecology, International Society of Pelvic Surgery and the National College of French Gynecologists-Obstetricians]. PMID- 10675829 TI - [Human papillomaviruses, cell cycle and cervical cancer]. AB - Human papillomaviruses (HPVs) are associated with a broad spectrum of cutaneous and mucosal lesions. Until now, more than 120 genotypes have been identified. Most HPVs are associated with benign lesions. Nevertheless certain HPV types are frequently found in carcinomas. For instance, HPV 16 and 18 which are frequently associated with cervical cancer, are capable of immortalizing and transforming primary keratinocytes. The mechanism of transformation is linked to the viral genome integration into the cell's DNA, accompanied by an overexpression of the E6 and E7 genes. The viral gene products interact with cellular proteins that regulate the cycle progression. In particular, the E6 protein binds to the p53 and the E7 protein binds to the p105(Rb). The inactivation of both cellular proteins distorts the cell cycle and results in genetic instability and cellular gene alterations. This article reviews the role of the viruses in the carcinogenesis, the genome structure and the gene expression of HPVs. It also addresses the cell cycle regulation with a focus on the role of HPVs in cell transformation. PMID- 10675830 TI - [Management of lymphoblastic lymphomas during pregnancy]. AB - Lymphoblastic lymphoma (non-Hodgkin lymphoma) is a highly uncommon but serious condition during pregnancy. With multidisciplinary management (obstetrics, pediatrics, hematology and anesthesia), outcome is generally good for both mother and child. Chemotherapy must be initiated rapidly, during pregnancy. Consequences depend on the stage of the disease, its progressive nature and the of pregnancy. During the first trimester, medical termination should be proposed in order to initiate chemotherapy cannot be started until the second trimester using alkaloids. Chemotherapy has little effect on the fetus during the second trimester. During the trimester, extraction should be discussed as soon as the fetal maturity is sufficient. PMID- 10675831 TI - [Effect of combined conjugated estrogen-medrogestone replacement therapy on lipid profiles, climacteric symptoms and the endometrium]. AB - OBJECTIVE: To assess the effects of continuous administration of conjugated estrogen combined with sequential administration of medrogestone on lipid profiles, climateric symptoms and endometrial tolerance. METHODS: This multicenter open study was conducted for one year to assess the effects of a hormone replacement therapy (HRT) regimen using Premarin (0.625 mg/day 28d/28) combined with medrogestone 5mg for 12 days (d17-d28 of each 28-day cycle) on lipid profiles, climateric symptoms and cycle control in 228 post menopausal women with an intact uterus. The subjects were recruited in 23 centers in 7 countries in Europe and Asia. Serum lipid/lipoprotein levels were determined at baseline and at cycles 3, 6, 13; endometrium biopsies were performed at screening then at cycle 13. Climateric symptoms and bleeding patterns were recorded by the patients from daily diaries cards collected at baseline and at visits during cycle 3, 6, 9, and 13. RESULTS: By cycle 3, the conjugated estrogen-medrogestone combination induced significant modifications of the lipid profile which were judged favorable. These modifications were maintained throughout treatment. All the baseline values were within normal limits. Mean variations compared with baseline values (expressed in mmol/l) after cycles 3, 6, and 13 were -0.46, 0.54, and -0.46 for total cholesterol (p<0.05), + 0.053, + 0.057, and + 0.078 for HDL-cholesterol (p<0.05) and -0.556, -0. 542, and -0.493 for LDL-cholesterol (p<0.001) respectively. VLDL-cholesterol levels were unchanged. Triglycerides increased significantly though moderately: + 0.12, + 0.15, and + 0.15 mmol/l at cycles 3, 6, and 13 respectively. Endometrial biopsies obtained at cycle 13 (n=195) did not reveal any endometrial hyperplasia. Withdrowal bleeding was predictable for a 6 to 7.4 day interval. The incidence of irregular bleeding varied from 7 to 33% and decreased progressively over the 13-cycle treatment. The incidence of amenorrhea increased from 14 to 52% over the 12 months studied. Finally, at each cycle, menopausal symptoms (mean number of hot flushes/day and Kupperman score) were significantly improved compared with the baseline. As expected, modifications were more pronounced after cycle 1, but improvements were maintained throughout the study. CONCLUSION: Continuous administration of Premarin in combination with sequential administration of medrogestone was found to be an effective treatment for menopausal symptoms. It was associated with favorable modifications of the lipid profile and was safe for the endometrium. PMID- 10675832 TI - [Hysterectomy for benign lesions in Brittany: analysis of medical practices]. AB - OBJECTIVES: By who, why and how are done hysterectomies for benign lesions. MATERIAL: and methods. The 413 medical files of all the patients who underwent an hysterectomy for benign lesion during the last trimester of 1997 were recorded by the Medical Information Departments of the 53 health establishments of the Brittany Region. Surgical procedures, medical indications, pathological findings were analyzed according to the guidelines encountered in the medical literature. RESULTS: Hysterectomies were done by many surgeons (112). Inaugural signs noted in the medical files were classical, but various and often associated without a main indication of hysterectomy. Histological diagnose were identical with those found usually in the literature. The abdominal route was mainly used, particularly when the operation was done by a generalist surgeon and in case of uterus weight superior to 250 g. The post operative outcome has revealed the same nature and frequency of complications as usually described. CONCLUSION: In this study, it appears that efforts remain necessary to clarify the indications for hysterectomy in the medical files (in order to promote the alternative procedures to the hysterectomy), and that the proportion of hysterectomies performed by the abdominal route should be reduced in aid of the others surgical routes. PMID- 10675833 TI - [Cervical adenocarcinomas: diagnostic, prognostic and therapeutic aspects in a 49 case-control study]. AB - OBJECTIVES: The aim of this study was to search for potential diagnostic, therapeutic and prognosis differences between a series of 49 adenocarcinomas of the cervix and a matched series of epidermoid carcinomas. METHODS: Forty-nine adenocarcinomas were treated between 1978 and 1992 and retrospectively compared to a series of 98 paired epidermoid carcinomas. RESULTS: The adenocarcinoma incidence is 5.4%. There was no significant difference for age distribution, parity, or hormonal status. There was also no significant difference for clinical features. Stage I appeared more frequently in the adenocarcinoma group (stage I: 69.4%, stage II: 14.3%, stage III: 14.3%, stage IV: 2%). Stage I are also more frequently found in the adenocarcinoma group (69.4% versus 42. 9%, p< 0,05). Combined radio-surgical treatment was proposed more often for the adenocarcinoma group (respectively radio-surgery combination 73%, radiotherapy alone 18%, surgery 9%); in the epidermoid carcinoma group, combined radio-surgical treatment and radiotherapy were the usual treatment (46%); surgery alone appeared in third rank place (8%). Adenocarcinoma pelvic recurrences appeared more frequently (28.6% for adenocarcinoma group versus 13.3% for epidermoid group p< 0.05), while distant recurrence was the same (12. 2% for adenocarcinoma group versus 11.2% for epidermoid group, p< 0. 05). Five years overall survival rate was worse for the adenocarcinoma group (52% versus 63.7%, p< 0.05) but the difference was not significant for the disease free survival rate. Only for stage Ib, there are also more pelvic recurrences (35.4% versus 13.1%, p< 0.05), more distant recurrences (9.6% versus 2.6%, p< 0.05), and lower overall survival for adenocarcinomas (58.7% versus 88.5%, p< 0. 01). CONCLUSION: The incidence of adenocarcinomas is slightly increasing (absolute value in our experience) and the low stages seem to be more frequent in our experience probably by staging inaccuracy. Adenocarcinoma prognosis seems to be worse because of its poor radio-sensitivity. It seems necessary to optimize clinical staging and therapeutic protocols excluding radiotherapeutic approach, including surgical purposes or radio-surgical associations if unfavorable histological features or tumoral enlargement (T> 3 cm) are found. PMID- 10675834 TI - [Risk factors for prematurity in France and comparisons between spontaneous prematurity and induced labor: results from The National Perinatal Survey 1995]. AB - OBJECTIVE: To study risk factors of total preterm delivery, spontaneous preterm delivery and induced preterm delivery. POPULATION: Representative sample of births in 1995 in France, including 12869 single births. METHOD: Preterm deliveries included all births before 37 weeks. Risk factors were analyzed with logistic regression. Factors of spontaneous preterm delivery (after spontaneous onset of labor) and factors of induced preterm delivery (after induction or cesarean section before labor) were compared with polytomous logistic regression. RESULTS: The main risk factors of preterm delivery were history of adverse pregnancy outcome (ORa=4. 5), history of induced abortion (ORa=1.5), 35 year old or more (ORa=1.5) and inadequate antenatal care (ORa=2.1). Other factors such as age under 20 or being single were not significantly linked to preterm delivery. Risk factors differed slightly between spontaneous and induced preterm deliveries. CONCLUSION: The present risk factors do not always correspond with the well-known factors. Thus the assessment of risk factors would be necessary at regular interval. The small differences between the risk factors of spontaneous preterm delivery and induced preterm delivery may be explained by difficulties in defining those two types of preterm delivery or by difficulties in distinguishing specific causes for each of them. PMID- 10675835 TI - [Metrorrhagia during the second trimester of pregnancy: obstetrical and perinatal outcome. A retrospective study including 85 cases]. AB - OBJECTIVE OF THE STUDY: To examine pregnancy outcome in patients with second trimester vaginal bleeding and to determine a relationship between presumed etiology and perinatal outcome. MATERIAL: and methods: A retrospective study performed in Toulouse La Grave CHU between January 1993 and December 1997 including 85 cases of vaginal bleeding from 15 to 27 week's amenorrhea (90 fetuses). Results are compared to overall deliveries during the same period (14941 deliveries). RESULTS: Mean age at diagnosis is 29.8 years (SD: 5.3 years). Vaginal bleeding in the midtrimester concern 0.57% of deliveries. Abnormal ultrasonographic findings are observed in 73% of patients (placenta previa, subchorionic hematoma, placental abruption) and 81% were hospitalized (mean hospital duration: 18 days). Perinatal mortality was 17.04% and preterm delivery rate 30.6% (vs 11% in the overall patients). Perinatal complications are significatively increased compared with the overall population especially if ultrasonographic examination was abnormal. Second-trimester placental abruption had the worse prognosis. On the other hand, perinatal outcome was comparable when the origin of bleeding was unknown and ultrasonographic examination normal. 41% patients underwent cesarean section. CONCLUSION: Preterm delivery, perinatal mortality and morbidity are increased in patients with second-trimester vaginal bleeding. The risk is higher when abnormalities are detected at ultrasonography making ultrasonography a useful tool for predicting perinatal outcome. PMID- 10675836 TI - [Materno-fetal platelet allo-immunization revealed by in utero intracerebral fetal hemorrhage: proposed management for the next pregnancy]. AB - Materno-fetal platelet allo-immunization causes fetal or neonatal thrombocytopenia and sometimes severe intracerebral bleeding. The HPA-1s antigen is most generally implicated. This accident can occur during the first pregnancy with a major risk of severe recurrence during the next pregnancy. These women require specific care in a specialized center although no consensus has been reached on management of second pregnancies. Proposed treatments include immunoglobulins and/or corticosteroids, fetal blood puncture and unique or iterative platelet transfusions. PMID- 10675837 TI - [Drug use during pregnancy: survey in 250 women consulting at a university hospital center]. AB - A prospective study of drug use in pregnancy was performed in the maternity outpatient department of a university hospital in Southwestern France. Two hundred and fifty pregnant women were selected at random and interviewed. Eighty four percent of the women reported drug consumption, with an average of two medications per week. Some factors were significantly associated with an increase in drug use: European origin, high level of education, medical history, alcohol consumption. The most commonly used drugs were iron (43% of the women), medications for venous disorders (22%) and gynecology (21%) and analgesics (19%). Chronic use (beginning before pregnancy) occurred in 9% of the women. Self medication accounted for around 20% of the women. Adverse effects were described by the women 25 times out of 544 exposures: they concerned 10 drugs and were not "serious". After delivery, 14 cases of malformations of the new-born, three stillbirths, 25 neonatal pathologies and 10 resuscitations were observed. Except one neonatal withdrawal syndrome after "in utero" exposure to benzodiazepines, no relationship between drug exposure and pregnancy outcome could be established. PMID- 10675838 TI - [Sickle cell anemia and pregnancy: review of 68 cases in Guadeloupe]. AB - Pregnancy in women with major sickle cell syndromes is a high risk maternofetal situation. This descriptive study presents the features and the clinical course of 68 pregnancies in sickle cell women who were delivered in Guadeloupe from January 1(st) 1993 to December 31(st) 1997. Specific complications were observed in all hemoglobin types, but with a severer course in SS women. Painful vaso occlusive crises were the main causes of hospitalisation (88% of SS pregnancies and 27% of SC pregnancies) associated most often with worsening anemia and / or infection. Acute chest syndrome was observed in all genotypes at any time throughout pregnancy and during the post partum period. One death occurred (a 16 years old SBeta(+)thal woman). Fetal mortality and morbidity were also high, intrauterine growth retardation and fetal death being the most frequent fetal complications. The rates of prematurity (21%) and caesarean section (48%) were higher than in the whole population. Three (3) neonatal deaths occurred. A multidisciplinary and specific approach, vigilance of health care providers and patient compliance are required to manage efficiently pregnancy, delivery and post partum in sickle cell women. PMID- 10675839 TI - [Length of stay in maternity wards after normal delivery: diverging point of views]. AB - OBJECTIVES: To compare maternity ward professionals' and patients' views regarding the length of stay in the maternity after a normal delivery and to explore working relationships with ambulatory health professionals. METHODS: Three surveys RESULTS: General professional agreement on a minimum of 4 days; few contacts with ambulatory health professionals. One third of the women found their length of stay excessive. Most did not appear to gain new skills after 3 days, nor to encounter major difficulties once they returned to their home. DISCUSSION: and conclusion. It will be essential to organize effective working relationships between the maternity ward and ambulatory health professionals to ensure appropriate follow-up after an earlier discharge from the hospital, in agreement with the capabilities and expectations of a majority of women. PMID- 10675841 TI - [In Process Citation] PMID- 10675840 TI - [Granulomatous recurrent mastitis during pregnancy]. AB - We report a case of granulomatous mastitis during pregnancy which raised important diagnostic and therapeutic problems. Several diagnoses were suspected and subsequently different therapeutic regimen were tried (antibiotic therapy associated with nonsteroidal antiinflammatory drugs, progestogens, vein tonicity drugs and bromocriptine). All were ineffective and the patients status worsened. Histology confirmed the diagnosis of granulomatous recurrent mastitis. Corticosteroid therapy and interruption of pregnancy successfully controlled the disease process. PMID- 10675842 TI - [In Process Citation] PMID- 10675843 TI - [Difficulties in antenatal recognition of fetal macrosomia]. PMID- 10675844 TI - [Obstetrical strategies and emergency procedures for delivery of macrosomic fetuses]. PMID- 10675845 TI - [The macrosomic newborn in the maternity ward: practical attitude]. PMID- 10675846 TI - [Obstetrical paralysis of the brachial plexus]. PMID- 10675847 TI - [Long-term consequences of fetal macrosomia]. PMID- 10675848 TI - [Management of pregnant women with chronic psychiatric disorders]. AB - This paper describes the psychic changes occurring within motherhood and their effects on the various kinds of mental disorders. The necessity of a multidisciplinary preventive and intervention network is emphasized. However, prenatal assessment is not predictive of what will happen postnatally. Only with the birth of the infant will the assessment of parenting begin. Some modification of the infancy and childhood protective regulations could help improving this post natal assessment of parenting ability. PMID- 10675849 TI - [Pre and postnatal depressions: importance of detection and care]. AB - It is highly important to recognize and care for pre and post delivery breakdown in order to relieve the mother's psychological pain and favor the ongoing motherhood process as well as the baby's psychological development which can be hampered by the rupture of mother child interactions during the early months of life due to the mother's depressive condition. PMID- 10675850 TI - Symptomatic Valgus Knee: The Surgical Options. AB - Valgus knee deformities requiring surgery are difficult to manage due to the relative rarity and abnormal biomechanics of the condition and the unique soft tissue and osseous pathologic features. Surgical options include arthroscopic debridement, abrasion arthroplasty, proximal tibial varus osteotomy, distal femoral varus osteotomy, combined femoral-tibial varus osteotomy, unicompartmental knee arthroplasty, and total knee arthroplasty. Each procedure has its own indications, contraindications, and limitations. PMID- 10675851 TI - Carpal Instability: Evaluation and Treatment. AB - Carpal instability is a common cause of wrist pain, motion loss, and disability. Diagnosis and treatment of carpal instability are dependent on a clear understanding of wrist anatomy and carpal kinematics, both normal and pathologic, as well as their relation to the current concepts regarding management. A brief review of anatomy and normal kinematics is presented, followed by a detailed discussion of specific instability patterns, including pathomechanics. A treatment algorithm is provided, detailing the authors' preferred treatment for the most common instability patterns. PMID- 10675852 TI - Total Joint Replacement: Optimizing Patient Expectations. AB - Rehabilitation of the patient who has undergone total hip or knee replacement embraces many facets of care, including prevention of complications, patient education, and a program of gradual resumption of normal functions. This program may be divided into three phases. In the perioperative phase, elimination of factors that contribute to morbidity will facilitate resumption of physical activities. In the interim phase (the first year following surgery), the patient's desire to return to full activities must be tempered by the goal of preserving for the longest possible time the mechanical-biologic construct of the joint replacement. Although a final functional result is usually achieved in the first 2 to 3 years following surgery, the patient must be followed up indefinitely. During this third phase of long-term assessment, the question of whether total joint arthroplasty was a success must be answered by the surgeon, by the patient, and by society. PMID- 10675853 TI - Glenohumeral Instability: Evaluation and Treatment. AB - Glenohumeral instability encompasses a spectrum of disorders of varying degree, direction, and etiology. The keys to accurate diagnosis are a thorough history and physical examination. Plain radiographs are frequently negative, especially in subtle forms of instability. Computed tomography (CT), CT arthrography, magnetic resonance imaging, arthroscopy, and examination under anesthesia may occasionally yield important diagnostic information. Nonoperative treatment of shoulder instability consists of reduction of the joint (when necessary), followed by immobilization and rehabilitative exercises. The length and the value of immobilization remain controversial. Rehabilitative programs emphasize strengthening f the dynamic stabilizers of the shoulder, particularly the rotator cuff muscles. Both arthroscopic and open techniques can be used for operative stabilization of the glenohumeral joint. Results of these repairs are assessed not only in terms of recurrence rate, but also in terms of functional criteria, including return to athletics. Some standard repairs have declined in popularity, giving way to procedures that directly address the pathology of detached or excessively lax capsular ligaments without distorting surrounding anatomy. Capsular repairs also allow correction of multiple components of instability. PMID- 10675854 TI - Percutaneous Lumbar Diskectomy. AB - The development of an approach for percutaneous lumbar diskectomy (PLD) began over 20 years ago. Since then, clinical investigations of manual and automated PLD techniques have recorded an average success rate of 50% to 70%. Currently, the indications for PLD include (1) a major complaint of acute unilateral leg pain localized to a single dermatome associated witha a single-disk herniation; (2) neurologic signs or symptoms appropirate to a single-disk herniation; (3) magnetic resonance imaginng, computed tomographic, or diskographic evidence of a single herniation contained within the annulus of the lumbar disk; and (4) failure of a well-managed course of conservative treatment to relive the pain and symptoms. Conventional laminotomy/laminectomy, with or without the use of a microscope or surgical loupes, remains the usual method of surgical care for symptomatic lumbar disk disease. The role of PLD awaits further prospective randomized controlled studies. PMID- 10675855 TI - Meniscus Tears: Treatment in the Stable and Unstable Knee. AB - Basic science research and follow-up studies after meniscectomy have provided convincing evidence of the importance of preservation of the meniscus in decreasing the risk of late degenerative changes. Whether in a stable or an unstable knee, if a meniscus tear cannot be repaired, a conservative partial meniscectomy should be undertaken to preserve as much meniscal tissue as possible. When feasible, repair should be carried out in young patients with an isolated meniscus tear, despite healing rates that are significantly lower than those obtained when meniscus repair is done with anterior cruciate ligament (ACL) reconstruction. The incidence of successful healing is inversely related to the rim width and tear length. In general, meniscus repair should be limited to patients under 50 years of age. Vertical longitudinal tears, including bucket handle tears, are most amenable to repair. Some radial split tears can be repaired. In an ACL-deficient knee, meniscus repair is more prone to failure if not performed in conjunction with an ACL reconstruction, and is not recommended. Meniscal allograft surgery is investigational but may hold promise for selected patients. PMID- 10675856 TI - Osteoporosis: The Role of the Orthopaedist. AB - Osteoporosis is one of the most prevalent musculoskeletal disorders encountered in orthopaedic practice today. This review provides an update on the pathophysiology of bone metabolism leading to osteoporosis, describes the latest methodology in the diagnostic workup of patients with low bone mass, and summarizes the current status of osteoporosis treatment regimens. The special needs of the osteoporotic fracture patient are also addressed. In general, load sharing devices and sliding nail-plate constructs are preferred over rigid internal-fixation systems. Prolonged immobilization should be avoided. PMID- 10675857 TI - Isolated and Combined Posterior Cruciate Ligament Injuries. AB - Posterior cruciate ligament (PCL) injuries represent 3% to 20% of all knee ligamentous injuries, but the diagnosis often is missed at initial evaluation. Diagnostic acumen is increased by knowledge of knee biomechanics and selective ligament-cutting studies. The examiner must differentiate the isolated PCL injury from combined ligamentous injury to determine appropriate treatment. Isolated acute PCL tears with less than 10 mm of posterior laxity at 90 degrees of flexion should be treated with an aggressive rehabilitative program. This amount of laxity is found in the majority of isolated acute PCL tears. Isolated acute PCL tears with more than 10 to 15 mm of posterior laxity and PCL tears with combined ligamentous injuries should be reconstructed. Large PCL bony avulsions should be fixed internally. Small PCL bony avulsions with more than 10 mm of posterior laxity should be reconstructed. Chronic PCL injuries initially should be treated with an aggressive rehabilitation program. If such a program is not successful in a patient with more than 10 to 15 mm of posterior laxity and no significant radiographic evidence of degenerative changes, the PCL should be reconstructed. PMID- 10675858 TI - Metastatic Tumors of the Spine: Diagnosis and Treatment. AB - Metastatic disease of the spine occurs in as many as 70% of patients with disseminated cancer and may result in vertebral collapse, spinal instability, and progressive neurologic compromise. Today, magnetic resonance imaging is the most effective means of differentiating benign from malignant causation of vertebral collapse, based on the imaging patterns and extent of marrow ablation. The more rapid the onset of the neurologic deficit, the worse the prognosis for recovery, no matter what treatment is instituted. The majority of vertebral lesions requiring decompression and stabilization emanate from the vertebral body and are best managed by anterior decompression and stabilization alone. With posterior element destruction, spinal subluxation through the involved segment, or involvement of the lumbar spine, a combination of both anterior and posterior stabilization is required. The author's preference is to perform anterior vertebral replacement with methylmethacrylate incorporating a Knodt distraction rod. This construct affords instantaneous stability that is not adversely affected by postoperative irradiation. Many devices can provide adequate posterior stabilization, but the author prefers to use Luque rods with sublaminar wire fixation. In a series of 77 patients with major neurologic compromise treated with this technique, 62% showed improvement by at least two Frankel grades, compared with fewer than 5% who improved after laminectomy decompression with or without irradiation. Nineteen of the 77 patients remained alive more than 4 years postoperatively. PMID- 10675859 TI - Tendon Disorders of the Foot and Ankle. AB - Attritional and traumatic injuries to the tendons around the foot and ankle are not uncommon. Treatment of overuse-type injuries (tendinitis) remains straightforward. However, surgical treatment of peroneal subluxation, Achilles tendon ruptures, and posterior tibial tendon insufficiency remains somewhat controversial. Generally speaking, soft-tissue reconstruction of the superior peroneal retinaculum is superior to bony procedures for peroneal dislocation. Open repair of a torn Achilles tendon is more predictable than closed treatment. Good clinical judgment is needed in determining the best treatment for posterior tibial tendon problems. The painful os peroneum syndrome is a newly described spectrum of posttraumatic conditions that may be the cause of lateral foot pain, which is frequently difficult to identify. PMID- 10675860 TI - Locked Femoral Nailing. AB - Locked intramedullary nailing has become the standard of care for most femoral fractures. Originally designed to prevent rotation and shortening in comminuted fractures of the midshaft, its application has been extended proximally and distally to nearly all femoral fractures from the lesser trochanter to the supracondylar area. Achieving a closed reduction and selecting the proper starting point in the piriformis region are crucial to a successful result. Following the proper surgical technique for the specific nail used is more important than nail material or design. Large-diameter reamed nails provide greater strength than unreamed nails. Static locking has been shown to yield nearly the same high union rates as dynamic locking and is now the accepted standard. Distal targeting of the interlocking screw remains the most difficult aspect of the surgical technique; most surgeons prefer freehand targeting with a sharp trocar. Second-generation (reconstruction) nails, with screws directed toward the femoral head, has extended the indications for locked nailing proximally to subtrochanteric fractures and combined femoral neck-shaft fractures. PMID- 10675861 TI - Elbow Arthritis: Treatment Options. AB - The treatment of elbow arthritis is conceptually similar to that for arthritis of other major joints. The treatment of elbow arthritis has been evolving rapidly due to advances in arthroscopic techniques and surgical treatment for contractures and improved prosthetic designs. The reliability of total elbow replacement is approaching that of total replacement of the knee, hip, and shoulder. There remain a number of controversies and unanswered questions that require further experience and longer follow-up for resolution. PMID- 10675862 TI - Plica: Pathologic or Not? AB - A fold that occurs within a joint is referred to as a plica synovialis. Three such plicae are seen with regularity within the human knee joint. These folds are normal structures that represent remnants of mesenchymal tissue and/or septa formed during embryonic development of the knee joint, and can be seen during arthroscopic inspection of the knee joint. Controversy exists within the orthopaedic community as to whether a plica can develop pathologic changes sufficient to cause disabling knee symptoms. The author defines the clinical syndrome, describes the arthroscopic appearance of pathologic plica, and outlines nonsurgical and surgical methods of management of this uncommon condition. PMID- 10675863 TI - Outcomes Research in Orthopaedics. AB - A new agenda in outcomes research has developed in the past decade. The stimulus has come as the result of rapidly increasing health care costs, marked variations in utilization of health care services, and deficiencies in the research literature. Outcomes research includes methods such as analysis of large databases, small-area analysis, structured literature reviews (meta-analysis), prospective clinical trials, decision analysis, and guideline development. Clinical research should be prospective and should employ modern statistical and assessment methods. The focus of this research is on patient-oriented outcomes of care rather than on assessments of the process of care. To illustrate these applications in orthopaedics, lumbar spine fusion with internal fixation for "spinal instability" is presented as an example. Completed large-database analyses, small-area variation studies, and a meta-analysis indicate the need for clinical studies. An outline of the form and content of such a study is presented. PMID- 10675864 TI - 1999 Costenbader Lecture. Outcome study in amblyopia: treatment and practice pattern variations. AB - This study retrospectively evaluates the results of amblyopia therapy and suggests hypotheses for future study. We address the various methods of treatment and evaluate the results from the most common therapy techniques. Practice pattern variations were analyzed in addition to the analysis of overall results. For ophthalmologists, there is a need to determine whether actual medical practice approaches the established standard of care, if it exists at all. How often are medical procedures, thought to be appropriate, based on anecdotal observation (case reports) rather than hard data (clinical trials)? The 3 types of vision loss evaluated were strabismic, anisometropic, and deprivation amblyopia. The methods of treatment studied were full-time patching part-time occlusion, penalization, and occlusion of the contact lens. Nine centers, thought to have private as well as indigent patients, were recruited to participate in this study. The centers responded by filling out an extensive questionnaire and sending the information through the World Wide Web for inclusion in a spreadsheet. This information was then collated, and various statistical programs tabulated the results. Although trends, as a consequence of therapy, are suggested from our retrospective analysis, concrete results can only arise from a randomized prospective study. The study included 279 patients. There were a similar number of male and female patients. Only 77% of the patients without fusion before treatment had either single binocular vision or peripheral fusion at the conclusion of therapy. The log improvement of vision was significant in each group. Factors that potentially influenced the results were severity of distance acuity in the amblyopic eye before treatment, duration of treatment, and length of daily patching. The paper suggests that worse vision, not better vision, at the beginning, predicts better outcome in terms of improvement of visual acuity. For example, visual acuity less than 20/70 at the initiation of treatment led to better visual results of geometric log improvement. Surprisingly, among the 9 centers studied, there was a statistically significant difference in many of the areas related to practice patterns. Patient compliance, which directly affects outcome, was highly variable and is a factor that may be readily influenced by the treating physician. PMID- 10675865 TI - Why does early surgical alignment improve stereoacuity outcomes in infantile esotropia? AB - PURPOSE: Recent studies of infantile esotropia suggest that early surgical alignment may enhance stereopsis and that alignment during the first 6 months of life may be optimal. Early surgery both establishes alignment during an early critical period for the development of stereopsis and minimizes the duration of misalignment. Here we examine the role of these 2 factors in promoting improved stereopsis outcomes. METHODS: Participants were 129 consecutive patients enrolled in a prospective study of infantile esotropia who were followed up for a minimum of 5 years. At ages 5 to 9 years, Randot stereopsis was evaluated. RESULTS: Multiple linear regression indicated that duration of misalignment, but not age at alignment or age at onset, was a significant factor in determining random dot stereopsis outcomes. Moreover, patients with stereopsis were less likely to have a loss of horizontal eye alignment requiring surgery than patients without stereopsis (14% versus 32%; z = 1.96, P =.05). Patients with stereopsis were also less likely to have dissociated vertical deviation than patients without stereopsis (25% versus 63%; z = 3.36, P <.001). CONCLUSIONS: The results suggest that early surgical alignment is associated with better stereopsis in those patients with infantile esotropia who were treated during the first 24 months of life, because early surgery minimizes the duration of misalignment, not because alignment is achieved during an early critical period of visual maturation. Random dot stereopsis can also be achieved in patients with alignment provided that the duration of misalignment is not prolonged. Improved outcomes of random dot stereopsis are associated with more stable long-term alignment outcomes. PMID- 10675866 TI - Factors influencing stereoacuity in accommodative esotropia. AB - PURPOSE: Despite successful optical realignment, many children with accommodative esotropia (ET) have abnormal stereoacuity. In a prospective study, we examined the influence of age of onset, age at alignment, duration of constant misalignment, and accommodative convergence/accommodation ratio on random dot stereoacuity outcomes in accommodative ET. METHODS: Participants were 111 consecutive children with accommodative ET. Random dot stereoacuity was measured using the Randot preschool stereoacuity test, the Randot stereoacuity test, the infant random dot stereoacuity cards, and the Lang 1. RESULTS: Age of onset has only a minor influence on stereoacuity (P <.02); children with onset >/=age 25 months have better stereoacuity compared with children with an onset between ages 7 and 17 months. Age at alignment has a minor influence on stereoacuity (P <.001); children with intermittent ET who have been treated have better stereoacuity than children with a constant ET aligned between ages 6 and 24 months and after age 24 months. Duration of constant misalignment has the strongest influence on stereoacuity (P <.001); children who had intermittent misalignment or who had a constant misalignment of less than 4 months' duration have better stereoacuity than patients who had a constant misalignment greater than 4 months' duration. The accommodative convergence/accommodation ratio does not influence stereoacuity outcomes (P >.10). CONCLUSIONS: Fine random dot stereoacuity is associated with a constant misalignment of less than 4 months' duration. These findings promote prompt and aggressive treatment of accommodative ET at the onset of intermittent or constant misalignment. PMID- 10675867 TI - The relationship between nystagmus and surgical outcome in congenital esotropia. AB - PURPOSE: Congenital esotropia is often associated with congenital nystagmus. This study examines the relationship between the presence of nystagmus and surgical outcome in the treatment of patients with congenital esotropia. METHODS: In this institutional retrospective study, we reviewed the charts of 200 consecutive patients who underwent surgical correction for congenital esotropia between 1991 and 1995. Preoperative clinical characteristics and subsequent need for additional strabismus surgery for a residual or consecutive deviation were noted. Minimum follow-up was 6 months after the original operation. RESULTS: Of the 84 patients who met the inclusion criteria, 15 patients (18%) had latent or manifest latent nystagmus, and 69 patients (82%) had no nystagmus. Eight of the 15 patients with nystagmus had or required reoperation according to our criteria (53%). Nineteen of the 69 patients (28%) without nystagmus had or required reoperation (P =.155). CONCLUSIONS: Nystagmus, when associated with congenital esotropia, may increase the risk of requiring additional strabismus surgery for residual or consecutive deviations. Appropriate and complete preoperative counseling of patients with congenital esotropia who also have nystagmus should include this increased risk. PMID- 10675868 TI - Oculographic and clinical characterization of thirty-seven children with anomalous head postures, nystagmus, and strabismus: the basis of a clinical algorithm. AB - BACKGROUND AND PURPOSE: We studied children with nystagmus who also had anomalous head postures and strabismus to determine the etiology of the conditions and present a diagnostic clinical algorithm. METHODS: The patients for this study were among the 560 patients evaluated in the ocular motor neurophysiology laboratory between the years 1991 and 1997. Clinical characteristics, infrared oculography data, and medical and surgical treatments were entered into a database for analysis. Oculography was performed on all patients according to a standard protocol, and data were stored and analyzed off-line. Etiology of anomalous head posture was determined with both clinical and oculography information. RESULTS: Thirty-seven children are the subjects of this report. The etiology of anomalous head posture was a "gaze null" due to congenital nystagmus in 23 (62%) patients, an "adduction null" due to manifest latent nystagmus in 12 (32%) patients, spasmus nutans in 1 (3%) patient, and strabismus in 1 (3%) patient. The patients' ages ranged from 9 months to 12 years and averaged 4.4 years. Sixty-nine percent were male patients. Nineteen (63%) of 30 patients had abnormal recognition (linear optotype) acuity in at least 1 eye on monocular cover; the recognition remained abnormal in 5 (17%) of 30 patients under binocular conditions. Thirty percent of patients had amblyopia, 16% had some structural disease of the eyes, 22% had some systemic syndrome or abnormality, 57% had a significant refractive error, and 27% had some ability to fuse. CONCLUSIONS: The major etiology for anomalous head posture in these patients was to adopt a gaze null due to congenital nystagmus (62% of patients) regardless of the direction of their anomalous head posture or type of strabismus. Moving the fixing eye as the first step for the anomalous head posture, combined with moving the nonfixing eye for the resulting strabismus may help treat these patients. PMID- 10675869 TI - Does primary intraocular lens implantation prevent "aphakic" glaucoma in children? AB - PURPOSE: Open-angle glaucoma may develop after surgery for congenital or developmental cataract with an incidence ranging from 3% to 41%. The pathogenesis of "aphakic" (open-angle) glaucoma remains unknown. Despite numerous reported clinical series (>1000 eyes), we are unaware of any reported case of open-angle glaucoma after primary intraocular lens (IOL) implantation for congenital or developmental cataract. We decided to test the hypothesis that primary posterior chamber IOL implantation might decrease the incidence of open-angle glaucoma in children. METHODS: Pseudophakic eyes were collected from surgeons who contributed data to a refractive study and who monitored intraocular pressure on a regular basis. IOL implantation was commonly performed in eyes with a corneal diameter >10 mm. Comparable primary data on aphakic eyes were included from 2 published studies on aphakic glaucoma, which included corneal diameters and the patient's age at surgery. Glaucoma-free survival estimates for each cohort were estimated. RESULTS: Only 1 case of glaucoma was found among 377 eyes with primary pseudophakia (mean age of patient, 5.1 +/- 4.7 years; mean follow-up, 3.9 +/- 2.7 years). There were 14 eyes (11.3%) with glaucoma among 124 aphakic eyes (mean age of patient, 2.7 +/- 2.6 years; mean follow-up time, 7.2 +/- 3.9 years). CONCLUSIONS: We report a decreased incidence of open-angle glaucoma among eyes rendered primarily pseudophakic compared with those that remained aphakic after cataract surgery. We propose 2 theories on the possible mechanism of reduction in the incidence of glaucoma in pseudophakic eyes. PMID- 10675870 TI - Heterochromia after pediatric cataract surgery. AB - PURPOSE: Changes in iris color have been noted anecdotally after cataract surgery in infants, but they have not been studied systematically. The mechanism for these iris color changes has not previously been reported in the biomedical literature. METHODS: Photographs were taken of both eyes of 15 children and 11 rhesus monkeys who had undergone unilateral cataract surgery. Masked examiners reviewed the photographs and compared the iris color of the eyes that were operated on with the eyes that were not operated on. Between 4 and 6 weeks postoperatively, the level of prostaglandin F(2alpha) in the aqueous humor (n = 4) and vitreous humor (n = 2) was measured in both the operated and nonoperated eyes of 4 monkeys that had undergone a neonatal lensectomy during the first 5 days of life. RESULTS: Thirteen of 15 children had a darker iris color in the operated eye in relation to the nonoperated (control) eye. Four of 11 monkeys had a uniformly darker iris in the operated eye; the other 7 monkeys had regional darkening or patches of darker iris in the eye that was operated on. The prostaglandin F(2alpha) levels in neonatal monkeys were higher in the aqueous humor and in the vitreous humor of the operated eye in relation to the nonoperated eye. CONCLUSION: In some children, cataract surgery is associated with a darkening of the iris color in the operated eye. We speculate that this darkening results from an exuberant prostaglandin release stimulated by the cataract surgery and may occur through the same or a similar mechanism by which latanoprost causes the darkening of iris color. PMID- 10675871 TI - Management and complications of congenital dacryocele with concurrent intranasal mucocele. AB - INTRODUCTION: The association of dacryocele and intranasal mucocele has been previously reported. Its incidence and optimal treatment are unknown. PATIENTS AND METHODS: A retrospective review of 22 patients with 30 dacryoceles was performed to determine the mean age at presentation, sex distribution, and prevalence of associated intranasal mucocele, associated dacryocystitis, and respiratory distress. The components of the examination, ancillary tests, treatment modalities, and treatment outcomes were then summarized. RESULTS: Unilateral dacryoceles were seen in 16 (73%) of the infants, and bilateral dacryoceles were seen in 6 (27%) of the infants. Four (25%) of the 16 patients who initially had unilateral dacryoceles later developed bilateral dacryoceles. Dacryocystitis, preseptal cellulitis, or both were present on presentation or developed in 18 (60%) of 30 dacryoceles. Nasal endoscopy was performed on 13 (59%) of 22 patients. Nasal examination with nasal speculum and headlight was performed on 7 patients (32%). A concurrent intranasal mucocele was diagnosed in 23 (77%) of 30 dacryoceles. Respiratory distress arose in 5 (71%) of 7 patients with bilateral intranasal mucoceles and in 2 (22%) of 9 patients with a unilateral intranasal mucocele. Thirty-four procedures were performed. Seven dacryoceles (21%) were treated with nasolacrimal duct probing under topical anesthesia. Another one (3%) was treated with needle aspiration with later definitive therapy. All other procedures were managed under general anesthesia. These included 2 nasolacrimal duct probings (6%), 2 probings with silicone tube placement (6%), 10 probings with intranasal mucocele marsupialization and silicone tube placement (29%), and 12 probings with marsupialization alone (35%). Two (29%) of the 7 probings performed under topical anesthesia failed, whereas all other procedures were successful. One dacryocele spontaneously resolved. CONCLUSIONS: Congenital dacryoceles are commonly associated with intranasal mucoceles, dacryocystitis, and preseptal cellulitis. Respiratory distress is common in bilateral cases. Bilateral nasolacrimal duct probing should be considered in unilateral cases because of the high incidence of occult contralateral involvement. PMID- 10675872 TI - Is screening for primitive neuroectodermal tumors in patients with unilateral retinoblastoma necessary? AB - Retinoblastoma is the most common childhood intraocular tumor, occurring in 1 of 18,000 live births. Retinoblastoma may occur as a germinal mutation or a somatic mutation. Forty percent of retinoblastoma cases are caused by a germline mutation and include those patients with a positive family history of the disease. Children with hereditary forms usually have multifocal, bilateral retinoblastoma, whereas children with the somatic form have unilateral, unifocal disease. However, up to 15% of cases of sporadic unilateral retinoblastoma may be hereditary. It is important to recognize that this subgroup of unilateral patients remains at risk for the development of second tumors as well as second primary tumors of the intracranial midline, or "trilateral retinoblastoma." We report a case of a 2-month-old child with unilateral retinoblastoma in whom pinealoblastoma subsequently developed. PMID- 10675873 TI - Anterior segment ischemia after strabismus surgery with microvascular dissection. AB - Anterior segment ischemia is a rare but well-known complication of extraocular muscle surgery.(1) Several surgical techniques have been used to prevent this complication in high-risk patients. A number of studies have suggested that microvascular dissection and preservation of the anterior ciliary vessels during strabismus surgery may reduce the risk of ischemic complications. (2-4) We present a case in which anterior segment ischemia occurred despite the use of this vessel-sparing technique. PMID- 10675874 TI - Strabismus surgery in children with Mobius syndrome. AB - Mobius syndrome is a congenital disorder of facial diplegia associated with lateral gaze paralysis. Although palsy of the sixth and seventh cranial nerves is the minimum diagnostic finding for Mobius syndrome, neuropathologic evidence indicates that this is a more complex syndrome.(1) Clinically, it is characterized by a total absence of facial expression and severe esotropia. Other anomalies may be associated with this syndrome, especially other cranial nerve palsies and Poland syndrome. The etiology of this syndrome has not been clearly established. Brain stem necrosis resulting from a vascular deficiency has been offered as a possible pathogenetic explanation.(2) The strabismus in Mobius syndrome is congenital esotropia with bilateral limitation in abduction. Even though many reports have described the various features of Mobius syndrome, only a few articles have reported the results of strabismus surgery in children, including bimedial rectus muscle recession. (3-5) Some authors report that bilateral medial rectus muscle recession alone has been disappointing; therefore, a combination of a medial rectus muscle recession and a lateral rectus muscle resection was recommended for satisfactory results. (5-7) In more severe cases, muscle transposition was needed to ensure straight position of the eyes in primary gaze. (8-9) PMID- 10675875 TI - Postoperative intraocular pressure elevation after the use of Healon GV in pediatric cataract surgery. AB - Intraocular pressure elevation after the use of viscoelastic agents in uncomplicated cataract surgery has been well documented in adults. However, pediatric patients are thought to clear residual viscoelastic agents from the anterior chamber more easily than adults, presumably because of healthier trabecular meshwork. (1) We report on a series of 4 eyes of 4 children with previously normal intraocular pressure who underwent cataract extraction with primary (3 patients) or secondary (1 patient) intraocular lens implantation with Healon GV, which was complicated by marked postoperative intraocular pressure elevation (greater than 30 mm Hg). The patients, aged 5 to 14 years, had an intraocular pressure ranging from 34 to 50 mm Hg with Tonopen or applanation tonometry 1 day, postoperatively associated with nausea, eye pain, and microcystic corneal edema. Viscoelastic material was not entirely removed during surgery. Each of these cases occurred after a change in our preferred viscoelastic agent from one with less viscosity to Healon GV. Medical management controlled the elevated intraocular pressure in all cases without affecting the visual outcome. However, 1 patient with intractable nausea and vomiting required hospitalization for rehydration. With meticulous removal of all viscoelastic material at the completion of surgery, we have not documented any additional cases of postoperative pressure elevation. PMID- 10675877 TI - Reply PMID- 10675876 TI - Congenital nystagmus: In search of simplicity on the other side of complexity. PMID- 10675879 TI - A Boston T party - The highlights. PMID- 10675878 TI - 'Cleaning' autologous bone marrow transplants with E. coli toxin. PMID- 10675881 TI - Histamine control of sleep, learning and memory. PMID- 10675880 TI - RNA as a small-molecule drug target - Letter to the Editor. PMID- 10675883 TI - Conopeptides: From deadly venoms to novel therapeutics. AB - Marine cone snails have developed many distinct venoms that contain biologically active peptides as part of an envenomation survival strategy for feeding and defense. These peptides, known as conopeptides, have been optimized through evolution to target specific ion channels and receptors with very high affinities and selectivities. Side effects of currently available therapies often arise from their lack of selectivity between pharmacologically relevant targets and targets that have a similar structure but different function. As conopeptides can be highly selective between closely related receptor subtypes, they could meet specific therapeutic needs with a reduced likelihood of side effects. PMID- 10675884 TI - Integrated bacterial genomics for the discovery of novel antimicrobials. AB - Sequencing of bacterial genomes has been progressing with breathtaking speed. Currently, the genomes of 23 bacterial species are sequenced, with approximately 40 more sequencing projects in progress. Industrial research is now facing the challenge of translating this information efficiently into drug discovery. This review will summarize the impact of bacterial genomics, bioinformatics and second generation genomic technologies on target identification, assay development, lead optimization and compound characterization. PMID- 10675885 TI - Stress proteins as targets for anti-inflammatory therapies. AB - Microbial heat shock proteins (HSPs) are ubiquitous and highly immunogenic. In healthy humans, B- and T-cells with specificity for self-HSP can be easily detected. In patients with chronic inflammatory diseases, raised levels of antibodies and T-cells with reactivity to self-HSP have been observed. Based on this and other evidence, this raised immune reactivity might be the result of stress-induced upregulation of self-HSP during inflammation and is possibly caused by tissue destruction. More importantly, immunization with conserved sequences of microbial-HSP increases resistance to the induction of autoimmune disease. Together, it appears that immune reactivity directed towards self-HSP can be part of a regulatory immune effector mechanism that contributes to maintenance of self-tolerance and has anti-inflammatory activity. Boosting of such anti-inflammatory effector mechanisms by artificial immunization offers attractive immunotherapeutic possibilities. PMID- 10675886 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- 10675887 TI - Combinatorial chemistry. PMID- 10675888 TI - A genome analysis production line. PMID- 10675889 TI - Genetic defects of the growth hormone-insulin-like growth factor axis. AB - Our understanding of the physiology of the growth hormone-insulin-like growth factor (GH-IGF) axis has been characterized by remarkable advances in the past decade, with clarification of genetic defects in the development of somatotropes, GH secretion and action, and IGF synthesis and action. Combined efforts of research in this area and the development of animal models of growth retardation have also indicated new genetic abnormalities that might prove to cause short stature in humans. Genetic defects, both established and hypothetical, are reviewed, and a pragmatic clinical approach to the genetic investigation of short statured patients is presented. PMID- 10675890 TI - Do cytoskeletal components control fatty acid translocation into liver mitochondria? AB - For two decades it has been assumed that inhibition of carnitine palmitoyltransferase I (CPT-I) by malonyl-CoA represents the main regulatory mechanism of liver ketogenesis. However, recent evidence indicates that CPT-I activity is also controlled by interactions between mitochondria and cytoskeletal components. This newly recognized mechanism emphasizes the emerging role of the cytoskeleton in the regulation of metabolic pathways. PMID- 10675891 TI - The new human kallikrein gene family: implications in carcinogenesis. AB - The traditional human kallikrein gene family consists of three genes, namely KLK1 [encoding human kallikrein 1 (hK1) or pancreatic/renal kallikrein], KLK2 (encoding hK2, previously known as human glandular kallikrein 1) and KLK3 [encoding hK3 or prostate-specific antigen (PSA)]. KLK2 and KLK3 have important applications in prostate cancer diagnostics and, more recently, in breast cancer diagnostics. During the past two to three years, new putative members of the human kallikrein gene family have been identified, including the PRSSL1 gene [encoding normal epithelial cell-specific 1 gene (NES1)], the gene encoding zyme/protease M/neurosin, the gene encoding prostase/KLK-L1, and the genes encoding neuropsin, stratum corneum chymotryptic enzyme and trypsin-like serine protease. Another five putative kallikrein genes, provisionally named KLK-L2, KLK L3, KLK-L4, KLK-L5 and KLK-L6, have also been identified. Many of the newly identified kallikrein-like genes are regulated by steroid hormones, and a few kallikreins (NES1, protease M, PSA) are known to be downregulated in breast and possibly other cancers. NES1 appears to be a novel breast cancer tumor suppressor protein and PSA a potent inhibitor of angiogenesis. This brief review summarizes recent developments and possible applications of the newly defined and expanded human kallikrein gene locus. PMID- 10675892 TI - Newer agents for hormonal contraception in the male. AB - Efforts to create a hormonal contraceptive for men use testosterone to suppress the production of pituitary gonadotropins and, hence, spermatogenesis. However, conventional testosterone must be administered by frequent injection, and when given alone, is not 100% effective. Therefore, newer androgens and agents that synergistically suppress gonadotropin production are being studied to create an effective and commercially viable contraceptive. PMID- 10675893 TI - Answering the question: 'why did biocatalysis in organic media not take off in the 1930s?'. PMID- 10675894 TI - Response from kvittingen PMID- 10675895 TI - On the optimization of classes for the assignment of unidentified reading frames in functional genomics programmes: the need for machine learning. AB - At present, the assignment of function to novel genes uncovered by the systematic genome-sequencing programmes is a problem. Many studies anticipate that this can be achieved by analysing patterns of gene expression via the transcriptome, proteome and metabolome. Thus, functional genomics is, in part, an exercise in pattern classification. Because many genes have known functional classes, the problem of predicting their functional class is a supervised learning problem. However, most pattern classification methods that have been applied to the problem have been unsupervised clustering methods. Consequently, the best classification tools have not always been used. Furthermore, the present functional classes are suboptimal and new unsupervised clustering methods are needed to improve them. Better-structured functional classes will facilitate the prediction of biochemically testable functions. PMID- 10675896 TI - Transgene-mediated modifications to animal biochemistry. AB - The application of gene-transfer technology to domestic animals provides a way for the introduction of genes encoding biochemical pathways that are currently nonfunctional in these animals. This might provide a mechanism for increasing the availability of specific substrates that currently limit certain production characteristics, such as the production of wool. The progress and problems associated with recent attempts to transfer a cysteine biosynthetic pathway and a glyoxylate cycle to sheep are discussed, in addition to the extension of this concept to other biochemical pathways. PMID- 10675897 TI - Cold-adapted enzymes: from fundamentals to biotechnology. AB - Psychrophilic enzymes produced by cold-adapted microorganisms display a high catalytic efficiency and are most often, if not always, associated with high thermosensitivity. Using X-ray crystallography, these properties are beginning to become understood, and the rules governing their adaptation to cold appear to be relatively diverse. The application of these enzymes offers considerable potential to the biotechnology industry, for example, in the detergent and food industries, for the production of fine chemicals and in bioremediation processes. PMID- 10675898 TI - Simulated moving-bed chromatography and its application to chirotechnology. AB - The increased awareness of the differences in biological activity of the two enantiomers of a chiral drug has raised the demand for enantiomerically pure products, particularly in the pharmaceutical industry. Simulated moving-bed chromatography can be used for the separation of the two enantiomers of a chiral molecule, which is feasible at all production scales, from laboratory to pilot to production plant. The use of non-enantioselective synthesis of racemic mixtures and simulated moving-bed enantiomer separation might make the development process of a new chiral drug substantially shorter and cheaper. PMID- 10675899 TI - Gene therapy: the first decade. AB - Gene therapy promises to revolutionize medicine by treating the causes of disease rather than the symptoms. We are nearing the end of the first decade of gene therapy, and this article summarizes the approaches taken, results achieved, lessons learned and important recent developments. The early results on the clinical efficacy of gene therapies were disappointing, largely because the available gene-transfer vectors proved to be inadequate. Recently, however, clinical benefit has been clearly demonstrated and great progress made in selecting and improving vectors. There is now every prospect that the second decade will see gene therapy live up to its enormous potential. PMID- 10675900 TI - Regulation of endocytic traffic by rho family GTPases. AB - Endocytosis is a complicated yet highly efficient process that involves the uptake and processing of cargoes, ranging from small molecules, to activated signalling receptors, to whole microorganisms. Regulation of endocytic pathways is poorly understood. Recent evidence suggests that the Rho GTPase family of signalling proteins is intimately involved in endocytic traffic, providing novel insights into the control mechanisms that govern this process. PMID- 10675901 TI - Germ cell cytonemes? PMID- 10675902 TI - Searching for a function for nuclear actin. AB - The abundant cytoskeletal protein actin has numerous cytoplasmic roles. Although there are many reports of the presence of actin in the nucleus, in general they have been discounted as artifactual. However, recent work has begun to provide evidence for important roles for actin in nuclear processes ranging from chromatin remodelling to splicing. In addition, several regulators of actin polymerization are localized to the nucleus or translocate to the nucleus in a regulated manner, suggesting that there is some function of actin in the nucleus that is subject to regulation. This review discusses the evidence for actin in the nucleus and summarizes recent work suggesting that actin or actin-related proteins are involved in the regulation of nuclear processes such as chromatin remodelling. PMID- 10675903 TI - Protein-only inheritance in yeast: something to get [PSI+]-ched about. AB - Recent work suggests that two unrelated phenotypes, [PSI+] and [URE3], in the yeast Saccharomyces cerevisiae are transmitted by non-covalent changes in the physical states of their protein determinants, Sup35p and Ure2p, rather than by changes in the genes that encode these proteins. The mechanism by which alternative protein states are self-propagating is the key to understanding how proteins function as elements of epigenetic inheritance. Here, we focus on recent molecular-genetic analysis of the inheritance of the [PSI+] factor of S. cerevisiae. Insights into this process might be extendable to a group of mammalian diseases (the amyloidoses), which are also believed to be a manifestation of self-perpetuating changes in protein conformation. PMID- 10675904 TI - Association of STATs with relatives and friends. AB - Members of the STAT family of transcription factors are present in species as diverse as mammals, insects and slime molds. Discovered as mediators of interferon-induced signals, the STATs were later shown to drive many different ligand-induced responses through receptor-induced tyrosine phosphorylation and dimerization. STAT1 also functions as a transcription factor, essential for the efficient constitutive expression of certain genes, without needing tyrosine phosphorylation, and phosphorylated STAT1 dimers mediate suppression - rather than activation - of some genes. STATs are present in the cytoplasm of untreated cells in multiprotein complexes, which might aid in their nuclear translocation and differential binding to DNA, thus contributing to the specificity of STAT action. This review explores the diverse protein-protein interactions that underlie the multiple functions of the STATs. PMID- 10675905 TI - Integrin signalling: a new Cas(t) of characters enters the stage. AB - Cellular morphology is determined by the organization of the intracellular actin cytoskeleton, which is influenced by external and internal cues. Focal adhesions are sites at which the actin cytoskeleton is linked to the extracellular matrix by integrin receptor complexes. In addition to providing structural tethering points for cells, integrin receptor complexes transduce signals that influence a broad range of cellular processes, including migration, proliferation, transformation and apoptosis. The Cas proteins (p130Cas, HEF1/Cas-L and Efs/Sin), a family of docking proteins containing multiple interaction domains, are important components of integrin receptor signalling and have been implicated in all of these processes. PMID- 10675907 TI - Careers-perspective interview. PMID- 10675906 TI - Interaction blues: protein interactions monitored in live mammalian cells by beta galactosidase complementation. PMID- 10675908 TI - The neurobiology of duchenne muscular dystrophy: learning lessons from muscle? AB - Several forms of inherited muscular dystrophy are associated with brain abnormalities and cognitive impairment. One of the most common and severe of these diseases is Duchenne muscular dystrophy (DMD). Dystrophin, the product of the DMD gene, is found in neurones, where it is associated with the postsynaptic membrane. Cognitive impairment in individuals with DMD is thought to be due to an abnormality in the neuronal membrane that is caused by lack of dystrophin. Recent experimental evidence has provided valuable clues in our understanding of the complex molecular neurobiology of muscular dystrophy. PMID- 10675909 TI - Cell birth, cell death, cell diversity and DNA breaks: how do they all fit together? AB - Substantial death of migrating and differentiating neurons occurs within the developing CNS of mice that are deficient in genes required for repair of double stranded DNA breaks. These findings suggest that large-scale, yet previously unrecognized, double-stranded DNA breaks occur normally in early postmitotic and differentiating neurons. Moreover, they imply that cell death occurs if the breaks are not repaired. The cause and natural function of such breaks remains a mystery; however, their occurrence has significant implications. They might be detected by histological methods that are sensitive to DNA fragmentation and mistakenly interpreted to indicate cell death when no relationship exists. In a broader context, there is now renewed speculation that DNA recombination might be occurring during neuronal development, similar to DNA recombination in developing lymphocytes. If this is true, the target gene(s) of recombination and their significance remain to be determined. PMID- 10675910 TI - Unveiling synaptic plasticity: a new graphical and analytical approach. AB - Short-term synaptic plasticity has a key role in information processing in the CNS, whereas memories can be formed through long-lasting changes in synaptic strength. Despite the importance of these phenomena, it remains difficult to determine whether a synaptic modulation is expressed at a presynaptic or postsynaptic site. This article describes a new approach that, in its simplest form, can identify the site of expression by direct graphical means. A more sophisticated form of the technique can quantify functional synaptic properties and determine which of these properties is altered following a modulation of synaptic strength. PMID- 10675911 TI - Peptide hormones and neuropeptides: birds of a feather. PMID- 10675912 TI - What is a neuropeptide? PMID- 10675913 TI - Reply to de Wied and Kastin. PMID- 10675914 TI - Immunization with beta-amyloid: could T-cell activation have a harmful effect? PMID- 10675915 TI - Cross-modal reorganization of human cortical functions. AB - Recent technological development has opened fascinating opportunities in research on cognitive functions of the human brain. For example, cortical representations of sensory functions and their reorganization, which have been studied thoroughly in animals, are far better understood in humans now than they were only a decade ago. Hemodynamic and electromagnetic studies have demonstrated that a modality specific brain area that is totally deprived of its normal sensory input becomes responsive to stimulation of other modalities. The functional significance of this cross-modal activation was recently indicated by, for example, studies showing that the occipital cortex of the blind is activated by sound changes, when the task is to detect these changes. Moreover, trans-cranial magnetic stimulation applied to the occipital cortex of blind individuals results in distortions and omissions of letters in Braille text being read by the subject. Contrary to prevailing views, cross-modal neural reorganization might, as shown by recent results, take place even in the mature human brain. PMID- 10675916 TI - Acetylcholine-mediated modulation of striatal function. AB - Striatal spiny neurones serve as a major anatomical locus for the relay of cortical information flow through the basal ganglia. these projection neurones also represent the main synaptic target of cholinergic interneurones, whose physiological role in striatal activity still remains largely enigmatic. The striatal cholinergic system has been implicated in the pathophysiology of movement disorders such as Parkinson's disease, but the cellular mechanisms underlying cholinergic-neurone function are still unknown. On the basis of in vitro electrophysiological evidence, obtained from a rat corticostriatal-slice preparation, we propose that endogenous ACh exerts a complex modulation of striatal synaptic transmission, which produces both short-term and long-term effects. ACh-mediated mechanisms might be of crucial importance in processing the cortical inputs to the striatum. PMID- 10675917 TI - The origin and migration of cortical neurones: new vistas. AB - The principal neuronal types of the cerebral cortex are the excitatory pyramidal cells, which project to distant targets, and the inhibitory nonpyramidal cells, which are the cortical interneurones. This article reviews evidence suggesting that these two neuronal types are generated in distinct proliferative zones. Pyramidal cells are derived from the neuroepithelium in the cortical ventricular zone, and use the processes of radial glia in order to migrate and take their positions in the cortex in an 'inside-out' sequence. Relatively few nonpyramidal cells are generated in the cortical neuroepithelium: the majority is derived from the ganglionic eminence of the ventral telencephalon. These nonpyramidal neurones use tangential migratory paths to reach the cortex, probably travelling along axonal bundles of the developing corticofugal fibre system. PMID- 10675919 TI - Rarity and diversity in Amazonian forest trees. PMID- 10675918 TI - Channel noise in neurons. AB - The probabilistic gating of voltage-dependent ion channels is a source of electrical 'channel noise' in neurons. This noise has long been implicated in limiting the reliability (repeatability) of neuronal responses to repeated presentations of identical stimuli. More recently, it has been shown to increase the range of spiking behaviors exhibited in some neural populations. Channel numbers are tied to metabolic efficiency and the stability of resting potential, and channel noise might be exploited by future cochlear implants in order to improve the temporal representation of sound. PMID- 10675921 TI - How the locust got its stripes: the evolution of density-dependant aposematism. PMID- 10675920 TI - Testosterone and maternal effects - integrating mechanisms and function. PMID- 10675922 TI - The calculus of biodiversity: integrating phylogeny and conservation. PMID- 10675923 TI - Transposable elements and host genome evolution. AB - Several recent reports have challenged the idea that transposable elements (TEs) are mainly 'selfish' or 'junk' DNA with little importance for host evolution. It has been proposed that TEs have the potential to provide host genomes with the ability to enhance their own evolution. They might also be a major source of genetic diversity, allowing response to environmental changes. Because the relationships between TEs and host genomes are highly variable, and because the selfish, junk and beneficial DNA hypotheses are by no means mutually exclusive, a single label for these relationships appears to be inappropriate and potentially misleading. PMID- 10675924 TI - Evolutionary consequences of dating the Yixian Formation. AB - The Yixian Formation of northeastern China has yielded important new fossils that are fuelling debates on the origin of angiosperms, on the early radiation of birds and of mammals, and on the origin of feathers. Although these fossils provide a wealth of detailed anatomical information, knowledge of the absolute age of the Yixian Formation is crucial if we are to understand their true evolutionary significance. The age of the Yixian Formation has been disputed, but recent evidence provides strong support for an Early Cretaceous rather than a Late Jurassic age. PMID- 10675925 TI - Colonization and diversification: towards a phylogeographic synthesis for the Canary Islands. AB - Recently, the Canary Islands have become a focus for studies of the colonization and the diversification of different organisms. Some authors have considered Canarian endemisms as relicts of Tertiary origin, but new molecular data suggest a general pattern of continental dispersion followed by in situ speciation. Recent phylogeographic studies are revealing variants of the simple stepping stone colonization model that seems to hold for many Hawaiian groups. Many factors can generate deviations from such a pattern: the stochastic nature of colonization, competitive exclusion, phylogenetic constraints on adaptive evolution and extinction. An understanding of island colonization and diversification can best be developed from an ecosystem level synthesis as more data for the Canarian archipelago come to hand. PMID- 10675926 TI - Recreating ancestral proteins. AB - Tracing the history of molecular changes using phylogenetic methods can provide powerful insights into how and why molecules work the way they do. It is now possible to recreate inferred ancestral proteins in the laboratory and study the function of these molecules. This provides a unique opportunity to study the paths and the mechanisms of functional change during molecular evolution. What insights have already emerged from such phylogenetic studies of protein evolution and function, what are the impediments to progress and what are the prospects for the future? PMID- 10675927 TI - A consistent equation for ecological sensitivity in matrix population analysis. PMID- 10675928 TI - Reply from T. Benton and A. Grant. PMID- 10675930 TI - Reply from G. Bernasconi and J.E. Strassmann. PMID- 10675929 TI - From the laboratory to the field: the advantage of pleometrotic colony founding. PMID- 10675931 TI - Jensen's inequality and optimal life history strategies in stochastic environments. PMID- 10675932 TI - Early tetrapod evolution. AB - Tetrapods include the only fully terrestrial vertebrates, but they also include many amphibious, aquatic and flying groups. They occupy the highest levels of the food chain on land and in aquatic environments. Tetrapod evolution has generated great interest, but the earliest phases of their history are poorly understood. Recent studies have questioned long-accepted hypotheses about the origin of the pentadactyl limb, the phylogeny of tetrapods and the environment in which the first tetrapods lived. PMID- 10675933 TI - [Acute acromioclavicular dislocations]. AB - Acromioclavicular dislocations represent over 10% of acute traumatic injuries to the shoulder girdle. The mechanism is usually a direct impact on the shoulder with the arm in adduction, producing rupture of the acromioclavicular (AC) ligaments, then of the coracoclavicular (CC) ligament, with displacement of the lateral end of the clavicle. Rockwood described 6 grades of injury. Physical examination usually provides the diagnosis, which is confirmed by radiological examination. X-rays centered on the AC joint, if necessary with forceful adduction of both shoulders or under traction, are useful to evaluate the severity of the lesion. Grade I and II lesions are usually treated conservatively by simply immobilizing the arm for 3 to 4 weeks. Surgical treatment is usually advocated for grade IV, V and VI lesions: AC or CC fixation, sometimes associated with ligament repair, depending on the surgeons. AC pinning or C-C screw fixation are the techniques most often used. Management of grade III lesions remains controversial. Some authors advocate immediate surgical treatment in young, active patients, in heavy laborers and even in slender individuals. The choice of the operative technique is controversial, as no single technique has clearly proved to be superior to others. Other authors advocate conservative treatment, which gives functional results which patients consider quite acceptable, with faster recovery; patients should be informed that results are essentially similar, whatever the treatment. The possibility of performing secondary operations with good results in cases with failure of conservative management is a further argument in favor of applying conservative therapy first in acute injuries. PMID- 10675934 TI - Elbow instability. AB - An understanding of elbow instability is predicated on knowledge of the anatomy of the lateral collateral ligament complex and of the mechanism and kinematics of elbow subluxation and dislocation. The lateral collateral ligament complex is the key structure involved in recurrent elbow instability and it is virtually always disrupted in elbow dislocations that result from a fall. The ulnar part of the lateral collateral ligament complex (also known as lateral ulnar collateral ligament) is the critical portion of the ligament complex securing the ulna to the humerus and preventing posterolateral rotatory instability. The kinematics of elbow subluxation and dislocation are a three dimensional coupled motion referred to as posterolateral rotatory instability in which the forearm rotates off the humerus in valgus/external rotation during flexion from the extended position. Elbow instability is diagnosed on clinical examination by the lateral pivot-shift test, the posterolateral rotatory apprehension and drawer tests and on radiographic examination by performing stress x-rays. While the lateral pivot shift test is difficult to perform, the posterolateral rotatory drawer test is much less difficult. The most sensitive test, however, is the posterolateral rotatory apprehension test. A positive apprehension test in a patient presenting with a history of recurrent painful clicking, snapping, clucking, or locking of the elbow should lead one directly to the suspected diagnosis of posterolateral rotatory instability. Treatment is surgical, by repair or reconstruction of the lateral collateral ligament complex, specifically the ulnar part. Deficiencies of the coronoid and/or radial head must be addressed. PMID- 10675935 TI - Ulnar variance and its relationship to ligament injuries of the wrist. AB - An anatomic study on cadavers and a clinico-radiological study on patients were undertaken to verify if ligament injuries of the wrist could be associated with ulnar variance. Neither scapholunate nor triquetrolunate ligament injuries could be related to shorter or longer ulnae. Longer ulnae were associated with significantly more perforations of the triangular fibrocartilage complex (TFCC). PMID- 10675936 TI - Intracarpal ligamentous lesions associated with fractures of the distal radius: outcome at one year. A prospective study of 95 cases. AB - Intracarpal ligamentous lesions associated with fractures of the distal radius (FDR) are frequent. The prevalence of these lesions has been assessed either by arthrography or by arthroscopy, but their outcome remains unknown. We carried out a radiographic study to assess the incidence of intracarpal ligamentous lesions with scapholunate (SL) and/or lunotriquetral (LT) dissociation and their outcome at one year. These lesions were termed "dissociative ligamentous lesions" (DLL). This prospective series consisted of 102 consecutive FDR's. The initial x-rays, immediate postoperative x-rays and x-rays at 1 year were studied. We studied the relationships of the bones of the first carpal row, abnormal joint space widening, Gilula's lines and the values of the intracarpal angles. The evolution of the carpal height ratio between day 0 and one year was studied. Complete xrays were available for 95 patients. There were 9 epiphyseal, 45 metaphyseal and 41 mixed fractures. DLL's were diagnosed in the early stages in 40 patients. There were 29 isolated SL lesions, 2 isolated LT lesions and 9 cases of associated SL and LT lesions. At 1 year, the diagnosis was confirmed in all these cases but a further case of SL dissociation was diagnosed. At 1 year, 61% of DLL's showed significant loss of carpal height and were considered as progressive. There was an association between the type of fracture and the presence or absence of DLL (p = 0.02). This study, based on radiographic analysis alone, showed 43% DLL's. The majority could be identified immediately. These findings are similar to those in recent arthrographic or arthroscopic studies, but the interest of plain radiographic study is to diagnose only those lesions having a definite effect on the carpus ("static instability"). At 1 year, 61% of lesions diagnosed have significantly affected carpal height. PMID- 10675937 TI - Soft tissue stabilization in the management of chronic scapholunate instability without osteoarthritis. A 15-year series. AB - Management of chronic scapholunate instability without osteoarthritis remains controversial. Some surgeons favor partial wrist arthrodesis; others, soft tissue stabilization. Many techniques for soft tissue repair have been described but with few or unpredictable results. We reviewed all our cases of scapholunate instability without osteoarthritis treated by soft tissue stabilization. Since 1979, 37 soft tissue stabilization procedures have been performed to correct dynamic (25) or static (12) scapholunate instability without osteoarthritis. The average time from injury to surgical treatment was 7.2 mos. (range 0.25 to 36 mos.). Three cases were treated within the first month of injury. The choice of repair was determined intraoperatively. The scaphoid shift must be easily reducible to make the case eligible for soft tissue repair. The scapholunate ligament was usually disrupted from palmar to dorsal, and the average amount of disruption was 74%. When scapholunate ligament remnants were of sufficient quality, secondary repair was performed; but if not, ligament reconstruction using tendon grafts or capsulodesis was performed. The procedures used were secondary ligamentous repair in 16 (by direct suture, reinsertion using anchor and/or transosseous reattachment), ligament reconstruction using tendon grafts in 6, capsulodesis in 7 and a combination of these procedures in 8. The mean follow up was 27 mos. (range 2 to 62 mos.). Postoperatively, there was an 83% decrease in pain. The average wrist motion was 60 degrees extension, 47 degrees flexion, 18 degrees radial deviation and 28 degrees ulnar deviation (92%, 84%, 106% and 88% of preoperative values and 88%, 75%, 78% and 76% of the uninvolved wrists, respectively), and the grip strength was 28 kg (117% of preoperative value and 78% of the uninvolved wrists). On roentgenograms, the mean static scapholunate distance was 4.2 mm (a 26% loss of reduction compared to the early postoperative gap), but scapholunate and radiolunate angles were within normal values (58 degrees and 9 degrees, respectively). At follow-up, one patient presenting a small zone of chondromalacia on the scaphoid at the time of secondary ligamentous repair developed severe radioscaphoid arthritis 15 months postoperatively. The results were further assessed according to the form of instability, delay before surgery, severity of disruption and type of repair. Patients with static instability showed worse clinical and radiological findings than those with dynamic instability. Surgical delay did not influence the outcome. The more severe the ligament disruption was, the poorer were the results. All types of repair had a comparable outcome except those treated by ligament reconstruction using tendon grafts. The results in the latter group were unsatisfactory in terms of motion, grip strength and radiological findings. This technique has been abandoned by the group. In conclusion, soft tissue stabilization is part of the armamentarium in the management of reducible chronic scapholunate instability without osteoarthritis. Ligament reconstruction using tendon grafts gave, in our hands, unsatisfactory results. Otherwise, all types of repair achieved a relatively pain-free wrist, with acceptable motion, grip strength, scapholunate and radiolunate angles but with a wider than normal static scapholunate distance. A longer follow-up is needed to assess the effect of this abnormal gap. Factors that favorably affected the outcome were: dynamic type of instability and partial disruption of the ligament. PMID- 10675938 TI - Blatt dorsal capsulodesis for scapholunate instability. AB - This retrospective series reviews 17 patients with scapholunate instability treated with the Blatt procedure between 1994 and 1997. Indications were 11 cases of preradiographic instability, three dynamic and three static instabilities. Subjective and objective assessment was carried out. Average pain and level of activity score was 60.8/80 (good). Only three patients failed to continue their jobs. Ten patients were fully satisfied, and seven had minor to major reservations. Flexion loss averaged 11 degrees and extension loss was 11.3 degrees. Grip force improved significantly by 11.2 kg. Associated scapholunate interosseous ligament repair in 6 patients resulted in no further improvement. Major complications were deep infection (one case) and reflex sympathetic dystrophy (two cases). Given the lack of a superior procedure, we considered the Blatt capsulodesis a valuable therapeutic option for cases of preradiographic and dynamic instability, or as an adjunct to scapholunate interosseous ligament repair in more acute lesions. PMID- 10675939 TI - The skier's thumb. AB - The incidence of skier's thumb (rupture of the ulnar collateral ligament of the first metacarpophalangeal joint) is increasing. To determine whether conservative or surgical treatment is indicated, ultrasound (US) and magnetic resonance imaging (MRI) have been advocated in the last few years. Surgery should be performed in the case of an unstable joint with a ligamentous tear or in the presence of a displaced bony fragment. Several techniques for surgical repair in acute and old ruptures are proposed. Conservative and postoperative treatment consists of immobilization of the joint in a splint or thumb spica cast for 4 weeks. The best results are obtained in bony avulsion fractures. Conservative treatment of lesions requiring surgical treatment may result in permanent disability of the joint; thus, correct diagnosis is mandatory. PMID- 10675940 TI - Arthroscopy of the shoulder. Current concepts review. AB - Arthroscopy of the shoulder has become much more common in the past decade as surgeons have developed proficiency with the arthroscope in the knee and appropriate instrumentation has been developed. In recent years arthroscopic techniques adapted to the shoulder have continued to evolve from a diagnostic to a treatment-oriented modality. It is now recognized and accepted as both a diagnostic and therapeutic technique in orthopedic surgery. A thorough knowledge of the anatomy, disorders, arthroscopic variations and pathological findings is essential to successfully perform the procedure. This paper discusses the operating room set-up, the portal placement and the indications for arthroscopy of the shoulder. PMID- 10675941 TI - Diagnosis and treatment of acute ruptures of the Achilles tendon. Current concepts review. AB - Subcutaneous rupture of the Achilles tendon seems to have become more common in recent years. This results from a combination of more awareness in the medical field and greater participation in physical activities by the general population. The causes of Achilles tendon rupture are multifactorial and still unclear. The diagnosis can be made based on physical examination; special diagnostic studies are rarely necessary. The literature on ruptures of the Achilles tendon and associated treatment has expanded over the past decade. The lack of a universal, consistent protocol for subjective and objective evaluation following treatment of Achilles tendon rupture has prevented any comparison of results. There is still controversy concerning the best treatment. From a literature review, it appears that a satisfactory outcome may be achieved with either nonoperative or operative treatment but surgical repair appears to provide better functional capacity. Lower rerupture rates and slightly improved strength and functional ability may be expected with surgical treatment; however, the rate of minor complications is higher than with nonoperative treatment. Reports in the literature indicate that in active, young, very demanding individuals, surgical repair should be considered, with nonsurgical treatment reserved for elderly or sedentary patients. There is no single, uniformly accepted surgical technique for Achilles tendon repair. Most acute ruptures have been treated successfully with simple end-to-end suture, although various augmentation procedures have been combined with simple suture with satisfactory outcomes. To minimize the complications typically associated with open surgery, percutaneous techniques to repair the ruptured Achilles tendon have been advocated, and the results are reported to be promising, although not without failures and complications. Several recent studies have reported functional benefits of early postoperative tendon mobilization in well-motivated patients, since treatment results are determined not only by the method of repair but also, and perhaps more importantly, by the early postoperative functional rehabilitation. PMID- 10675942 TI - Mechanisms of retrolisthesis in the lower lumbar spine. A radiographic study. AB - The study investigates lower lumbar segments with posterior vertebral shifts (retrolisthesis) with respect to the orientation of facet joints, disc height, lordosis of the lumbar spine, and orientation of vertebral endplates. Standing lumbar radiographs as well as CT and/or MRI investigations of 69 patients were analyzed. Data from patients with retrolisthesis (20 cases) were compared to data from patients with degenerative spondylolisthesis (DS, 23 cases), and from patients without signs of vertebral shifts (26 cases). The orientation of facet joints in segments with retrolisthesis was not different from segments without shifts, whereas the facet joints in patients with DS were oriented more sagittally. The overall lordosis of the lumbar spine and the endplate inclination were considerably reduced in patients with retrolisthesis, especially compared to those with DS. Disc height was comparable in retrolisthesis and DS, but was reduced compared to segments without shifts. The results support biomechanical considerations, that a retrolisthesis of a lower lumbar spine segment is correlated with a reduction of lumbar lordosis, endplate inclination, and segmental height. PMID- 10675943 TI - Treatment of intracapsular fractures of the femoral neck in Denmark: trends in indications over the past decade. AB - A questionnaire survey was set up in Denmark in 1996 including 40 orthopedic departments and 20 departments of general surgery, all dealing with the treatment of intracapsular fractures of the femoral neck. The aim of the survey was to investigate whether the treatment of these complex fractures in Denmark followed the international standard, the "gold standard", recommended in the recent international literature. A shift in the treatment was noted, as compared with an earlier questionnaire survey in 1988, with more orthopedic departments performing a graduated treatment with respect to the age of the patients and fracture grade (Garden class). That is: a) nondisplaced fractures, b) displaced fractures (b.1 below 75 years and b.2 above 75 years). Cannulated screws/pins were more commonly used in Garden I and II fractures (non displaced fractures) and in Garden III and IV fractures (displaced fractures) in patients below 75 years. Hemiarthroplasty/arthroplasty were more commonly used in the older age group, above 75 years, in displaced fractures (Garden III-IV). It is concluded that a shift in the treatment of these fractures has occurred, especially in orthopedic departments. One reason for this may be an increasing number of orthopedic specialists with experience in arthroplastic surgery, making it possible to perform and/or supervise younger surgeons in this procedure. Another reason must be an increasing awareness among orthopedic specialists in Denmark that complication rates in osteosynthesis of the displaced fractures (Garden III-IV) have been too high. PMID- 10675944 TI - A method to measure acetabular cup anteversion after total hip replacement. AB - The authors propose a simple and practical method to measure radiologically the angle of ante- or retroversion of the acetabular cup using a goniometer. It only necessitates an anteroposterior radiograph centered on the femoral head and another one centered on the public symphysis. Special x ray equipment, compass, conversion table, mathematical formulas, or a pocket calculator are not required. The opening of the prosthetic cup is projected on the film as an ellipse. According to the rules of descriptive geometry, the true size of the angle of anteversion is easily obtained. The geometric constructions consist in drawing four lines. The adequate positioning on a hip radiograph of the protractor, drawn on the goniometer, permits the direct reading of the true and planar anteversion angles of the cup. PMID- 10675945 TI - [Study of the posterior cruciate ligament using a 3D computer model: ligament biometry during flexion, application to surgical replacement of the ligament]. AB - We have developed a 3D computed model of the knee joint, constructed from MRI acquisitions in a living individual. We have used this model to perform an anatomic and biometric study of the posterior cruciate ligament (PCL) during flexion, and an assessment of the optimal location for an intraarticular graft. The method used a 3D computed model constructed from MRI acquisitions during knee flexion (0 to 75 degrees). The range of motion was limited by a positioning device. We took 13 acquisitions from 0 to 75 degrees of flexion. Each acquisition consisted of 21 sagittal cross sections of 3 mm slice thickness. We used the Delaunay reconstruction to obtain a 3D geometric model. A matching process to fix one part of the articulation during the movement, allows for the kinematic analysis of the tibia relative to the fixed femur. This model allows to follow the displacement of a bone point during knee flexion. Knowing the relative displacement of the bone insertions of the ligament, it may be possible to determine the length of the PCL and its bands, to evaluate the length variation during movement, and to determine the optimal location for the insertion of an intraarticular graft, that would lead to the least stretch during flexion. It was found that the mean length of the PCL was 30.2 mm, with the posterior band being 30% longer than the anterior band. During flexion the posterior band increases its length by 10% at 50 degrees flexion, and by 20% at 75 degrees flexion. The anterior band stretches more, to reach 40% elongation at 75 degrees flexion. The best position for insertion of a graft seems to be in the posterolateral portion of the anatomic tibial insertion, and posterior to the anatomic femoral insertion. This method confirms the data in the literature, states precisely the length of the different bands of the PCL, and specifies the points of insertion for a graft, which lead to the least variation in length during flexion. PMID- 10675947 TI - Radial tunnel release and tennis elbow: disappointing results? AB - In a retrospective study, 19 patients with 20 decompressions of the posterior interosseous nerve for radial tunnel syndrome were reviewed. The results were evaluated using Roles and Maudsley's criteria; they were found to be consistent with those previously reported: i.e. 75% favorable outcomes. Despite this finding only 8 patients (40%) stated they were satisfied. The main reason was residual pain. Shorter delay between the onset of symptoms and surgical treatment as well as simultaneous release of the lateral epicondylar muscles was found to influence positively patient satisfaction. These findings suggest that the scoring system used in the present study and also in previous studies is inappropriate. They also cast some doubt on the role of compression of the posterior interosseous nerve in the pathogenesis of chronic lateral elbow pain. PMID- 10675946 TI - [Gauthier's subcapital osteotomy in the treatment of metatarsophalangeal luxation of the 2nd ray. Apropos of 44 cases with 5 year followup]. AB - The authors report on a series of 44 metatarsophalangeal dislocations of the second ray which were treated surgically using Gauthier's technique. The patients were 44 middle-aged women. The surgical indication was a dislocation of the 2nd metatarsophalangeal joint with hallux valgus. There was excess length of the second metatarsal ray or acquired shortness of the first metatarsal. The treatment always included an osteotomy of the neck maintained by a transosseous pin. The average follow-up was 8 years and 3 months (minimum 5 years). Postoperative results were evaluated using clinical and radiological criteria. Surgical treatment gave 68.2% very good and good results and 4 recurrences of dislocation. The results in this series are identical with those in other series reported, but the backward displacement of the head of second metatarsal was found to be limited. Weil's osteotomy seems to provide better results because it better restores the relative lengths of the metatarsals and often makes interphalangeal arthroplasty unnecessary. Gauthier's metatarsal osteotomy is an easy procedure which effectively improves static metatarsalgia, but it provides limited metatarsal shortening. Weil's osteotomy is preferable in cases with long lateral metatarsals. PMID- 10675948 TI - [Pneumatocyst of the sacrum. Apropos of a case]. AB - We report a new case of asymptomatic pneumatocyst of the sacrum in a 45-year-old man with a history of lumbago. Pneumatocysts are rare benign osseous lesions filled with gas and always found in the subchondral bone of the sacral or iliac side of the sacroiliac joint. The diagnosis is made by CT scan. In the sacral location, the pneumatocyst is asymptomatic and surgical treatment is not justified. PMID- 10675949 TI - Stress fracture of the acetabular roof. Case report and value of MRI. AB - The authors report the case of an 11-year-old boy with pain in the left leg without history of recent trauma. The diagnosis of a stress fracture of the acetabulum was made based on MRI and bone marrow biopsy. They discuss the role of MRI in the diagnosis of a stress fracture. PMID- 10675950 TI - Primary hydatid disease in lumbar muscles. AB - The authors report a case of primary hydatid disease in the lumbar muscles of a 40-year-old male patient. The rarity of this disease in our regions and the low incidence of this location make primary diagnosis difficult. The tumor had been treated elsewhere five years previously by means of simple excision. Recurrence of the lesion was diagnosed five years after the first surgery. Wide excision of the cyst and pericyst with a 3.5-cm security margin was performed. Six years after the last surgery, no recurrence has been detected. PMID- 10675951 TI - Bilateral intraosseous lipoma of the calcaneus. A case report. AB - A case of bilateral intraosseous lipomas of the calcaneus is presented. The bilateral localization of calcaneal intraosseous lipomas is extremely rare. PMID- 10675952 TI - Treatment of a simple bone cyst of the calcaneus by endoscopic curettage with cancellous bone injection. AB - The authors report a case of simple bone cyst involving the calcaneus, treated by curettage under endoscopy with cancellous bone injection, and its course and follow-up at two years. This new technique has not yet been published for simple bone cysts of the calcaneus. Endoscopic curettage of the cavity of a simple bone cyst can be advocated for the calcaneus to minimize incisions and to avoid cutaneous complications. PMID- 10675953 TI - Idiopathic symmetrical shortening of the fourth and fifth metacarpal and metatarsal bilaterally. A case report. AB - A case of idiopathic bilateral symmetrical shortening of the fourth and fifth metacarpal and metatarsal bones in an active 10-year-old Caucasian female is described. The deformity did not result from trauma or an endocrine disorder and it was not hereditary. The function of the hands and feet was normal, and the only discomfort was of a cosmetic nature. Metacarpal or metatarsal lengthening therefore seemed unnecessary. PMID- 10675954 TI - [Decision making in borderline situations--anesthesiological aspects]. AB - Over the last few decades, major biomedical developments have been taken place so that dying and death are nowadays more a matter of deliberate decision--a change that has profound ethical and legal implications. This progress has influenced medical decision-making generally and intensively, especially that of the surgeon and the anaesthesiologist. The representatives of these professions are often confronted with problems of life-sustaining therapy at the beginning and the end of life and of resuscitative measures. The surgeon and the anaesthesiologist have to accept the necessity of close partnership while maintaining a clear dividing line between their responsibilities, but at the same time jointly doing their utmost for the good of the patient. Above all the physician has to give due consideration to the patient's will, but there are many and sometimes great variations in the individual situations of conscious or permanently unconscious patients. The highest courts in Germany have laid down that the principles of medical ethics must supplement the law. PMID- 10675955 TI - [General anesthesia or spinal anesthesia for hip prosthesis replacement? Studies of acceptance of both procedures by patients]. AB - Patients undergoing total hip replacement are given general anaesthesia or spinal anaesthesia. The aim of this study was to investigate the experiences of patients before, during and after general anaesthesia (68 patients) or spinal anaesthesia (77 patients). Our investigation revealed that with regard to complications (nausea and vomiting, headache and back pains), no differences between the two methods occurred. Between 25 and 30% of the patients in both groups had these complications, although there were differences between both groups regarding their concomitant diseases and medication. Patients with spinal anaesthesia had a three times higher incidence of cardiac concomitant diseases and received corresponding drugs more frequently. We found that the time of postoperative analgesia after spinal anaesthesia (210 minutes) was significantly longer than after general anaesthesia (90 minutes). The majority of the patients in both groups (approximately 90%) were satisfied with the chosen method of anaesthesia and with the postoperative pain therapy. These findings make it possible to conclude that with the exception of differences in the postoperative analgesia time, there are no differences between general anaesthesia and spinal anaesthesia regarding complications and satisfaction of the patients with both methods of anaesthesia. PMID- 10675956 TI - [Continuous spinal anesthesia in very elderly patients with high anesthesia risk in traumatologic-orthopedic and general surgery interventions]. AB - Continuous spinal anaesthesia (CSA) was carried out via a 28-gauge spinal catheter in 154 surgical patients at the Department of Anaesthesiology and Critical Care at Radeberg Asklepios-ASB Hospital between May 1992 and March 1999. The method was used preferably in patients aged over 70 (mean age 82.3 years) with high general risk during anaesthesia (ASA III-IV) who underwent orthopaedic or general surgery of the lower limb and hypogastrium. Remarkably, an anaesthetic level of between Th 8 and Th 10 was achieved with the low initial dose of 7.5 mg of 0.5% hyperbaric bupivacaine. Only minimal cardiovascular and respiratory side effects were observed in comparison to single shot spinal and general anaesthesia. In the whole series, no anaesthesia-related complications were seen. Another benefit of CSA is the option of applying a second dose with longer duration of surgery to keep the optimal anaesthetic level. In addition, the method is suitable for postoperative analgesia over a period of 2 to 3 days. PMID- 10675957 TI - [High-frequency jet ventilation for placing tracheal stents--a case report]. AB - Stenoses of the larynx and trachea may cause acute life-threatening situations. Surgical procedures in patients presenting this type of problem are a real challenge for the surgeon and the anaesthesiologist. Depending on the extent and the nature of the stenosis, the insertion of a stent may be the best therapeutic option. In this case, the high frequency jet ventilation offers certain advantages for the surgeon. Thanks to modern jet ventilators with automatic pressure monitoring and jet ventilation tubes with a separate lumen for pressure monitoring, the danger of barotrauma is considerably reduced, even in patients with a high-degree stenosis of the larynx and trachea. During insertion of a tracheal stent during jet ventilation, the complete cross-section of the trachea must at least be temporarily available to the surgeon. In addition, at the end of the operation the newly implanted stent should not be altered by manipulations necessary for artificial respiration. We describe a new method which uses tracheal jet ventilation for implanting a stent with only short interruptions of artificial ventilation. During recovery from anaesthesia, there is no risk of dislocating the newly placed stent. PMID- 10675958 TI - [Tuberous breast syndrome. Report on a series of 22 operated patients]. AB - The tuberous breast syndrome is a rare unilateral or bilateral breast malformation presenting at the age of mammary development. It requires surgical correction, depending on the severity of the clinical expression, because of its inaesthetic appearance. We report a series of 22 patients (35 breasts) treated and followed up in the plastic and reconstructive surgery department of Rennes over a period of nearly ten years. The average age was 18 +/- 3.2 years (range: 15-26 years old). Long-term results were assessed by the surgical team and the patients on the basis of objective and subjective criteria with an average follow up of 36 months (12 to 116 months). In our opinion, surgical correction should be proposed after puberty with, whenever possible, section of the basal fibrous ring and glandular plasty via a periareolar incision. The use of mammary implant alone or in combination with breast tissue remodelling must be reserved for hypoplastic cases only. PMID- 10675959 TI - [Adipocyte microinfiltration in the face or tissue restructuration with fat tissue graft]. AB - The author presents his experience of the autologous fatty tissue transplantation. He calls his method microinfiltration of adipocytes (MIA). The fat of the body is harvested with the technique of syringe liposuccion without trauma and is refined and placed in an intricate layering of autologous fatty tissue with anatomic patented microcanulaes. This procedure named Lipostructure by Coleman is not a lipofilling or "liposculpture" as described by P. Fournier in 1989. By focusing on the procedure as autotransplantation fat grafting technique and fully respecting the delicate nature of the fat cell, the author has been able to document a high rate of success with 3 years follow-up. PMID- 10675960 TI - [Deep vertical lift and its development regarding the central facial area and lower two-thirds of the neck. Our technique]. AB - Deep vertical facelift, that we have developed over the last eight years, presented certain weak points in the central facial zone and lower two-thirds of the neck. Ageing of the central facial zone is associated with two main sings: skeletization of the orbit and accentuation of the nasolabial fold. These two elements are due to ptosis of the submalar fat. The obvious solution is to restore the position of this fat. We present a rapid and effective solution. In the lower two-thirds of the neck, in the presence of marked ptosis, we now opt for extensive deep vertical facelift with retroauricular dissection. The skin of the neck in front of the sterno-cleido-mastoid muscle is not dissected, but the SMAS is extensively dissected over the mandibular angle continuous with the facial dissection as fat as the lower part of the neck. PMID- 10675961 TI - [Skin and muscle flaps to cover and fill chronic open osteitis of the tibia. Results with 10 cases]. AB - Chronic osteomyelitis is a severe long-term bone infection which retains its mechanical qualities. The authors report 10 cases of osteomyelitis of the tibia treated by muscular and fascio-cutaneous flaps and reviewed at one year follow up. Four cases concerned the third upper part of the tibia, 3 the middle and 3 the lower third. Two total failures at the third lower part and three complications which finally healed with delay were observed. The results of this small series compared with the reports in the literature suggest the value of large excision with coverage by a well vascularized flap and the need for antibiotics. The choice of flap is related to type and site of the bone defect. Another question should be raised concerning the surgical strategy in one--or two -stage management and the duration of antibiotic therapy. PMID- 10675962 TI - [Position of venous outflow: a crucial aspect in flaps with 2 opposed pedicles. Application to the cutaneous delay phenomenon. Experimental study with rats]. AB - In the present study, the authors decided to focus on the effect of the venous outflow location in the survival of a flap with a choke vessels barrier by means of a rat bipedicled ventral island flap. In a first experiment, the extent of flap necrosis was examined in three experimental groups: group 1, alternate vein and artery ligation (n = 5); group 2, unilateral artery ligation (n = 5); group 3, unilateral pedicle ligation (n = 5). Less than 10% necrosis occurred in group 1; 28% in group 2 and 33% in group 3. In a second experiment we applied the first results to the delay phenomenon. 10 rats were divided into 2 groups: group 4, primary ligation of a complete pedicle on one side; group 5, primary alternate vein and artery ligation and the flaps were raised 21 days later. We observed 20% necrosis in group 4 and only 4% in group 5. The data of the first experiment indicate that a vein remaining near the artery can be a dramatic obstacle to the efficiency of choke vessels. The second experimental study shows the superiority of primary selective ligation in the delay phenomenon. PMID- 10675963 TI - [Facial aging. Analysis of esthetic semiology and proposal for systematic treatments]. AB - Surgical literature is severely lacking in anatomo-clinical descriptions of facial aging. Only a few authors provide minor reports on the pathophysiology of aging in order to propose methods of correction. It therefore appears important to start with a diagnosis of the lesions as precise as possible in order to be able to perform the most appropriate treatment. We propose a semeiological analysis of the face, not only stage by stage but layer by layer, starting with the skin, then the fat, muscle and bone. We will then consider the methods which appear to be the most suitable for correction of the various structures concerned by aging within the framework of a total facial reorganisation. We will conclude with a presentation of surgical cases describing the surgical techniques used, with respect to a semiological analysis from each case. PMID- 10675964 TI - [Unusual breast scars]. AB - The authors report the case of an unusual form of skin tattoo, discovered on the breasts of a young woman, corresponding to several scars forming a symbolic image, performed deliberately with a burning object. This form of mutilation is called "branding". Imported from England, it is developing in France as a result of fashion, but its followers could one day regret the permanent scars left by this deep burn. PMID- 10675965 TI - [Who are the French plastic surgeons?]. AB - Based on data from the national advice of the Conseil National de l'Ordre des Medecins (French Medical Board), the author found 654 plastic surgeons in France and classified them into five categories, demonstrating the increasing proportion of females in this specialty. PMID- 10675966 TI - [New frontiers in aesthetic surgery. Dallas, May 14-19, 1999]. PMID- 10675967 TI - Perceived, actual, and seasonal changes in the shape of the face, hands and legs. AB - In this study, we measured the shape of the face, legs, hands and fingers during the course of a day to determine the amount of swelling. We examined the relationship between the perception of swelling and the degree of actual swelling, and considered the influence of seasonal factors. The topology of the face was measured using the 3D curved shape measuring apparatus, VOXELAN, while the circumference of the legs and fingers and the volume of the hands were also recorded. The measurements were used to determine the amount of change in each parameter, which was then used to determine the degree of swelling. The subjects for the experiment were 10 healthy Japanese women aged 24 to 30 years of standard build (BMI: 19.3-25.0). Measurements were carried out twice a day in the mornings and afternoon, first between 8:30 and 10:00 a.m. and then between 4:00 and 5:30 p.m. At each measurement session, subjects were asked if they perceived swelling to have occurred. We investigated the relationship between the degree of actual swelling and the reported perception of swelling. We also investigated the influence of seasonal factors by conducting the same tests on the same subjects in summer (August 1997) and in winter (February-March 1998). The relationship between perceived and actual swelling differs depending on the part of the body. For the face, actual swelling correlates strongly with perceived swelling. This trend is particularly noticeable for the upper eyelids. For the thigh and lower leg, on the other hand, there was no significant difference. The frequency with which subjects reported the perception of swelling varied depending on the area of the body, and was generally extremely low for the thighs, hands and fingers. With respect to seasonal variation, swelling in the face, hands and feet tended to be more pronounced during the summer. In the facial region, the biggest difference was in the lower eyelid, where swelling increased more than five times. This level of variation suggests that the atmospheric temperature is the main factor affecting swelling. PMID- 10675968 TI - Blood pressure and hormonal responses to short whole body cold exposure in subjects with high dietary salt intake. AB - The objective of the present study was to test a hypothesis that a high dietary salt intake potentiates a cold induced increase in blood pressure in normotensive men. Male subjects (n = 12) were given 7 g day-1 sodium chloride during the cold months of the year, divided in 3-4 doses per day and dissolved in water, for 14 days additional to their normal diet which contained on the average 9.7 g sodium chloride per day. The same subjects, having their normal diet, served as controls. The resting blood pressure was measured on the fourteenth day seven times at the intervals of five minutes in a climatic chamber in thermoneutral conditions. Then the subjects, wearing a three-layer winter clothing, moved into a wind tunnel (-15 degrees C, air velocity 3.5 ms-1) in which they stayed for fifteen minutes and the blood pressure was recorded at the intervals of three minutes. After the cold exposure, the subjects moved back into the climatic chamber for 30 min and the blood pressure was measured as before the cold exposure. Blood samples were drawn before and after the experiment for ion and hormone measurements. A 12 h urine sample was collected just prior to the cold exposure. A significant difference both in systolic (7 mmHg) and in diastolic (7 mmHg) blood pressure was found between a salt load group and control group under thermoneutral conditions, repeatedly measured over 30 min (paired Student's t test; p < 0.05). During the whole body cold exposure, blood pressure significantly increased both with and without the extra salt load (repeated measures ANOVA, Student-Newman-Keuls; p < 0.05). The level to which the mean arterial pressure increased during the exposure was independent of the salt intake and the profile of the mean arterial pressure curve was similar in both groups. The systolic pressure increased by a 25 mmHg in both groups during the cold exposure. The increase in the diastolic pressure was significantly (paired Student's t-test, p < 0.05) higher in the high salt group (18 +/- 4 mmHg) than in the control group (12 +/- 3 mmHg) thus supporting partly our hypothesis. After the two-week high salt intake, serum Na+, K+, Cl-, Hct, and plasma Hb were at the similar level as before the extra salt intake. Plasma renin activity, NT-proANP, ANP, and serum aldosterone were not different between the groups, both before and after the cold exposure. The main findings are: 1) the mean arterial pressure increases to the same level and in the same manner independent of the salt load during a short whole body cold exposure and 2) in cold the diastolic blood pressure increases significantly more in people under a very high salt diet. PMID- 10675969 TI - Changes in physical characteristics and performance of elite sailors following introduction of a sport science programme prior to the 1996 olympic games. AB - The objective of this study was to examine changes in sailors' physical characteristics during three different time periods immediately before the 1996 New Zealand Olympic trials, as a result of a newly introduced sport science programme. Twenty five (19 male and 6 female) Olympic development squad members volunteered as subjects and completed fitness tests at different times between April 1995 and March 1996 after being administered with individualised physical training programmes. Statistically significant improvements were observed in body weight, sum of skinfolds, flexibility (assessed using a sit-reach test), aerobic endurance (assessed using a maximal effort 2500 m rowing test) and strength (assessed as the maximum number of push-ups, pull-ups, and sit-ups that could be completed in 2 minutes) over the three time periods. Thus, physical training was effective in improving many aspects of sailors' fitness, especially early in the sailing season as a result of pre-season training. Physical performance correlated poorly with both light and heavy wind racing performance. The results suggest that individually tailored training programmes will increase sailing specific fitness. However, it is impossible to know what proportions of racing performance can be attributed to physical fitness, skill, talent, and technology, therefore the effect of physical training on racing performance is difficult to determine. PMID- 10675970 TI - Analysis of heart rate variability during mental task with reference to ambient temperature. AB - The purpose of this study was to evaluate the cardiac autonomic control over mental task under various ambient temperatures (21 degrees C, 28 degrees C and 35 degrees C). Seven healthy male subjects engaged in the mental tasks, which consisted of distinctive reaction-time tasks. Respiratory coefficient of variation of instantaneous heart rate (CVRESP), derived from the cross correlation function between heart rate and respiratory curve, was used as a parameter to assess parasympathetic nervous functions. The difference between total coefficient of variation (CVIHR) and CVRESP was used as a parameter to assess sympathetic nervous functions. The mean heart rate increased at high ambient temperature (35 degrees C) and also during mental task. Both the effects of ambient temperature and task conditions were significant on heart rate, and also on CVIHR. Moreover, the effects of ambient temperature and task conditions in CVIHR were divided into the effect of ambient temperature on CVRESP and the effect of task conditions on the difference between CVIHR and CVRESP. These results implied that respiratory modulated parasympathetic activity might control basal the effect of ambient temperature, and the other components including sympathetic activity contribute to the increase in heart rate due to mental task. PMID- 10675971 TI - Effects of mental task on heart rate variability during graded head-up tilt. AB - In this study, we used spectral analysis of heart rate variability (HRV) to estimate the changes in autonomic control in response to disparate stimuli produced by mental task and graded head-up tilting. The low frequency (LF) component of HRV provided a quantitative index of the sympathetic and parasympathetic (vagal) activities controlling the heart rate (HR), while the high frequency (HF) component of HRV provided an index of the vagal tone. We studied 17 healthy male subjects (21-25 yr of age) who were placed on a tilt table and the graded tilt-protocol involved tilted sine angles 0.0, 0.2, 0.4, 0.6, 0.8, and 1.0. These tilt-protocols were repeated with or without the mental task, which consisted of auditory distinctive reaction-time tasks. The basal autonomic mode against the graded head-up tilt was characterized by reciprocal changes in sympathetic and vagal tones. There were significant increases of HR corresponding to the mental task with lower tilt-angle, albeit the changes with higher tilt angles were not significant. Furthermore, there were increases and decreases of the LF component induced by the mental task at lower and higher tilt angles, respectively. These results revealed that the different responses of HR and LF component against the same tasks could be derived from the alterations of autonomic mode during gradual changes in autonomic control. PMID- 10675972 TI - Cardiovascular regulation during water immersion. AB - Head-out water immersion at thermoneutral temperature (34-35 degrees C) increases cardiac output for a given O2 consumption, leading to a relative hyperperfusion of peripheral tissues. To determine if subjects immersed in water at a colder temperature show similar responses and to explore the significance of the hyperperfusion, cardiovascular functions were investigated (impedance cardiography) on 10 men at rest and while performing exercise on a leg cycle ergometer (delta M = approximately 95 W.m-2) in air and in water at 34.5 degrees C and 30 degrees C, respectively. In subjects resting in water, the cardiac output increased by approximately 50% compared to that in air, mainly due to a rise in stroke volume. The stroke volume change tended to be greater in 30 degrees C water than in 34.5 degrees C water, and this was due to a greater increase in cardiac preload, as indicated by a significantly greater left ventricular end-diastolic volume. Arterial systolic pressure rose slightly during water immersion. Arterial diastolic pressure remained unchanged in 34.5 degrees C water, but it rose in 30 degrees C water. The total peripheral resistance fell 37% in 34.5 degrees C water and 32% in 30 degrees C water. Both in air and in water, mild exercise increased the cardiac output, and this was mainly due to an increase in heart rate. Since, however, the stroke volume increased with water immersion, cardiac output at a given work load appeared to be significantly higher in water than in air. The arterial pressures did not decrease with water immersion, despite a marked reduction in total peripheral resistance. These results suggest that 1) during cold water immersion, peripheral vasoconstriction provides an additional increase in cardiac preload, leading to a further increase in the stroke volume compared to that of the thermoneutral water immersion, 2) the mechanism of cardiovascular adjustment during dynamic exercise is not changed by the persistent increase in cardiac preload in water immersion, and 3) a relatively high cardiac output during water immersion is to maintain a proper arterial pressure in the face of reduced vascular resistance. PMID- 10675973 TI - Protecting workers with developmental disabilities. PMID- 10675974 TI - Useful bookmarks for your virtual library. PMID- 10675975 TI - Control of exposure to perchloroethylene in commercial dry cleaning (ventilation). National Institute for Occupational Safety and Health. PMID- 10675976 TI - Identification of potential hazards associated with new residential construction. AB - There were several advantages and limitations of this observational study. The most important advantage of this study was the opportunity to observe residential construction workers performing their jobs. By observing work practices, valuable information was gathered about specific trades and their potential exposure to various chemical and physical agents. This information will be useful in guiding subsequent exposure assessments. Probably the greatest limitation of this study was the lack of participation by homebuilders. Ideally, observations of construction processes would have been more objective if the study included the participation of more than one homebuilder. Aside from one worker who was observed to wear safety glasses, leather gloves, and a dust mask, virtually no personal protective equipment (PPE) was observed onsite. Often small contractors do not have the financial resources necessary to procure the appropriate PPE and issue these items to the workers. Based on hazard prevalence, professional judgement, and the degree of hazardous product use, potential exposures that warrant quantitative sampling efforts during Phase 2 of this study are: bulldozer/backhoe operators--noise, vibration, diesel exhaust; concrete workers- naphtha, mineral spirits, Portland cement; asphalt workers--petroleum hydrocarbons, asphalt, mineral spirits; plumbers--methylethyl ketone, acetone, tetrahydrofuran, cyclohexanone; drywall finishers--total and respirable dust, hexane, acetone; painters--ethylene glycol, VOCs; masons--dust (during the preparation of mortar); floor preparation technicians--total and respirable dust; and ceramic tile installers--toluene, naphtha, silica (from grout powder). PMID- 10675977 TI - Self-assessment of exposure--a pilot study of assessment of exposure to benzene in tank truck drivers. AB - Occupational hygienists or safety engineers perform exposure assessments, mostly with very little participation by the workers. The objective of our study is to involve the workers themselves in the assessment and measurement procedure, the self-assessment method (SAE). A pilot study has been carried out involving tank truck drivers at a company transporting gasoline. The drivers were supposed to decide themselves when, and how often, they wanted to measure benzene exposure by using diffusive samplers that were then sent by mail for analysis. After every measurement they received their own results in a personal document for interpretation. The company management also received a document, which summarized all the drivers' measurements. Expert measurements, with the same type of sampler, were also accomplished to evaluate the self-assessments. The geometric mean and the 95 percent confidence intervals of the measurements made by the drivers (29 measurements) was 0.17 (0.11-0.26), and by an occupational hygienist (8 measurements) 0.12 mg/m3 (0.04-0.37). The results show that the drivers technically can perform SAE. Interviews with the workers and the management indicated that some kind of organizational support within the company is needed to implement the method into the regular internal control of the working environment. PMID- 10675978 TI - Chemical hazards in radiology. AB - A variety of chemicals are used in medical imaging as developer and fixer ingredients, germicides, and cleaning agents. Glutaraldehyde, a potent sensitizer, may cause occupational skin and respiratory diseases in exposed individuals. Poor ventilation, unsafe practices, and lack of hazard recognition may contribute to occupational asthma and other respiratory disease in susceptible medical imaging personnel. Failure to respond effectively to initial health complaints and reduce exposure levels can have serious consequences for affected employees. It is therefore important for occupational safety and health professionals to alert health facility managers to potential dangers and to recommend effective intervention strategies. When problems are identified, a multidisciplinary team approach is the best method for evaluating and controlling hazards. This team should include industrial hygienists, safety staff, occupational medicine physicians, mechanical and ventilation engineers, personnel specialists, and medical imaging staff. A thorough hazard assessment, medical diagnosis, and administrative personnel actions are critical to effective problem identification and correction. In the case of chemical sensitization, removal of the affected employee may be necessary. By working with designers and equipment installers to monitor compliance with appropriate codes and manufacturers' specifications, hazards can be prevented. We present additional operations, ventilation, and design improvements to reduce chemical exposures to radiology employees. PMID- 10675979 TI - Containment testing of laboratory hoods in the as-used condition. AB - Airborne contaminants generated inside laboratory fume hoods during use leak into the breathing zone of the user. Concentration of the leakage is unknown and variable depending on laboratory design, work practices, arrangement of internal apparatus, face velocity, and sash height. Surrogate tracer gas tests have been developed using sulfur hexafluoride (SF6) and a manikin to estimate leakage. This study presents results of hood leakage tests using SF6 with a manikin and then a live operator performing a phenol:chloroform (P:C) extraction. Four hoods were tested in each of three institutions during normal work hours with the lab occupied. The purpose of the study was to determine leakage concentrations for the SF6-manikin with effects of sash height, hood contents as found and after being cleaned out, face velocity, and the actual P:C and SF6 exposure concentrations of the user during work. Results indicate P:C was not detectable in the breathing zone of the 12 operators (< 0.1 ppm) at their selected operating sash heights (7 to 15 inches). Simultaneous SF6 concentrations were also minimal (average 0.06 ppm). Average leakage was 0.02 percent for SF6 and less than 2 percent based on chloroform concentrations measured in the breathing zone of the operator and inside the hood. SF6 percent leakage was greater when sash height was above the breathing zone of the manikin (average 2.09 percent) and lower leakage (average 0.02 percent) when below the breathing zone (26 inches or less). Average face velocity did not appear to be a predictor of average hood leakage. Cleaning out the hoods did not reduce leakage in most tests. The data from this study shows that when providing training on proper work practices for lab hood use, lowering the sash should be stressed as being the major factor in reducing hood leakage. PMID- 10675980 TI - Lead abatement training for underserved populations: lessons learned. AB - An environmental-justice (equity) grant program was used to make accessible an existing lead-training program to minority persons and residents of low-income communities. The purpose of the program was to enhance the knowledge base within the communities concerning lead hazards and intervention strategies and expand possibilities for employment in the lead abatement industry. Barriers to attendance were anticipated and addressed, and included transportation, meals, license application fees, reminders of course date and location, and day care. The program was evaluated through measures of recruitment rates, pre- and post testing scores, and change in perception of confidence at pre-test, post-test, and at four-month follow-up. Fee-paying registrants over the same time period were used as a comparison group. First day attendance rates for individuals recruited into the equity-grant was 59 percent, of these 94 percent completed all days. Equity and fee-paying groups had similar scores on the pre-test (p = .209), while mean scores on the final exam differed significantly (p < .001) between the groups and were 77 percent and 85 percent, respectively. After adjusting for demographic and course type attended, perceptions of self-efficacy (benefit) and outcome-effectiveness (confidence) increased significantly from pre- to post tests for both groups and remained at post-course levels at four months follow up. Lessons learned include: (1) Lead abatement and other related activities can be successfully taught through traditional training methods; (2) A necessary element for delivery of educational services to minority groups is forming workable ties with local community groups, but eligibility requirements must be maintained; (3) Once barriers to first-day attendance are overcome, the information necessary to perform specific work skills can be taught; (4) Positive changes in belief are not dependent on minority status, income, or education levels; (5) Training and education increased confidence in ability to perform learned skills, and belief that there will be a beneficial outcome when performed for themselves, their families, and communities; and, (6) A consensus regarding applicability of regulations must be achieved among federal, state, and local communities. PMID- 10675981 TI - Optimization of a solid sorbent dynamic personal air sampling method for aldehydes. AB - A solid sorbent personal dynamic air sampling method for aldehydes using chemisorption with 20 percent (w/w) O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) on Tenax TA has been optimized for several aldehydes. The method for formaldehyde was developed after optimization for valeraldehyde and acrolein. The effects of temperature, intermittent exposure, and flow rate on sampling efficiency were investigated. Vapors of known concentrations were generated in Tedlar gas bags by syringe injection. Aldehyde chemisorption by reaction with 200-mg coated solid sorbent in a Pyrex Tube (7-cm length, 7-mm OD, and 5-mm ID) occurred during dynamic collection with a personal sampling pump operated at 10, 50, or 100 mL/min. The oxime derivatives were 81-87 percent desorbed by shaking with hexane for two minutes or more, resulting in coefficients of variation ranging between 4-7 percent. Aliquots were then injected for gas chromatographic analysis on a nonpolar capillary column with mass spectrometric or 63Ni electron capture detection. Formaldehyde at 8 ppm hours equivalent to the permissible exposure limit (PEL) concentration was sampled with a recovery of 94 +/- 4 percent at 50.0 +/- 0.5 mL/min. Valeraldehyde and acrolein at their PELs showed no significant differences (P < 0.05) in recoveries at different relative humidities (1 and 90%), temperatures (1, 25, and 40 degrees C), or intermittent sampling protocol. The sampler capacity at 75 percent recovery was at a PFBHA:aldehyde molar ratio of 12:1 at 10.0 +/- 0.1 mL/min, 17:1 at 50.0 +/- 0.5 mL/min, and 26:1 at 100 +/- 1 mL/min. PMID- 10675982 TI - An effective protection factor study of respirators used by primary lead smelter workers. AB - An industrial hygiene study was conducted at a primary lead smelter to determine the effective protection factors for negative pressure and powered air purifying half-mask respirators. The study involved 99 paired personal samples taken in six different work areas, in which randomly chosen subjects from the workforce wore dual sampling pumps connected to closed-face 37-mm cassettes. The cassettes were attached either to the workers' lapels for exposure measurements outside the respirator or to a ported respirator for exposure measurements inside the respirator. Samples were collected throughout the work shift and analyzed for lead according to National Institute of Occupational Safety and Health (NIOSH) Method 7082. Using particle size distribution data obtained for the same workplaces, the within-mask samples were corrected for sampling bias. The negative pressure half-mask respirators showed a mean effective protection factor of 6.5 and a mean corrected effective protection factor of 4.56, with a 5th percentile less than 0.5. Approximately 80 percent and 90 percent of the effective protection factors and the corrected effective protection factors, respectively, were equal to or less than the assigned protection factor of 10. For the powered air purifying half-mask respirators, the means for effective protection factor and corrected effective protection factor were 18.20 and 11.92, respectively, with a 5th percentile of 1.0 or less. Approximately 90 percent and 95 percent of the effective protection factors and the corrected effective protection factors, respectively, were equal to or less than the assigned protection factor of 50. The uncorrected and corrected within-mask lead concentrations for both types of respirators exceeded the Occupational Safety and Health Administration (OSHA) permissible exposure limit (PEL) for lead by 19 percent to 58 percent. These results indicate that the straight application of assigned protection factors to actual workplace situations may not always be appropriate. PMID- 10675983 TI - Synthesis and cytotoxicity of 2,4-disubstituted and 2,3,4-trisubstituted brominated pyrroles in murine and human cultured tumor cells. AB - The 2,4-disubstituted and 2,3,4-trisubstituted brominated pyrroles were successfully prepared and demonstrated potent cytotoxicity against the growth of suspended murine and human tumors, i.e. leukemia and lymphomas, acute monocytic leukemia, and HeLa-S3 uterine carcinoma. The brominated compounds were more selective in inhibiting the growth of tumors derived from human solid tumors. Nevertheless, activity with some of the derivatives occurred in the human KB nasopharynx, SW-480 colon, and HCT ileum adenocarcinoma, and lung A549 carcinoma screens. In Tmolt4 T cell leukemia cells DNA synthesis was reduced over 60 min from 25 to 100 microM followed by RNA synthesis reduction. De novo purine synthesis was retarded with the regulatory enzyme PRPP-amido transferase being markedly inhibited with less effects on the activities of IMP dehydrogenase, dihydrofolate reductase,, and the nucleoside kinases. After 60 min incubations d[TTP] and d[GTP] pools were marginally reduced. In vitro ct-DNA studies suggest that the agents may affect the DNA molecule itself with increased DNA viscosity and the Tmolt4 studies suggest that DNA cross-linking of DNA strands may be present. PMID- 10675984 TI - Structure-activity relationships of polycyclic aromatic amines with calcium channel blocking activity. AB - 8-Benzylamino-8, 11-oxapentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecane (1) inhibits the calcium current in L-type calcium channels. A series of nitrobenzylamines (2, 3, 4), methoxybenzylamines (5, 6, 7), methylpyridines (8, 9, 10), and a phenylhydrazine derivative (11) of 8,11 oxapentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecane was synthesized. By substituting the 8,11-oxapentacyclo-[5.4.0.0(2,6).0(3,10).0(5,9)]undecane skeleton with 3-hydroxyhexacyclo-[6.5.0.0(3,7).0(4,12).0(5,10).0(9,13)]tridec ane (12), 8,13-dioxapentacyclo[6.5.0.0(2,6).0(5,10).0(3,11)]tridecane- 9-one (13), and pentacyclo-[5.4.0.0(2,6).0(3,10).0(5,9)]undecane (14), the effect of the polycyclic skeleton could also be investigated. Increased inhibition of calcium current was observed with aromatic substitution (especially ortho and meta substitution) in the pentacycloundecane series. The calcium channel activities of the methoxy compounds were slightly higher than those of the corresponding nitro compounds while a definite decrease in activity was observed for the phenylhydrazine and aminomethylpyridine derivatives. Increased inhibition of the calcium current was also observed for structures in which the polycyclic 'cages' were enlarged. Structure-activity relationships in this series of compounds therefore appear to be dominated by geometric or steric constraints. PMID- 10675986 TI - New NO donors with antithrombotic and vasodilating activities, Part 28. N-(1 cyanoalkyl)-N-hydroxyureas. AB - Nineteen N-(1-cyanoalkyl)-N-hydroxyureas comprising aliphatic (3a-i, 4a, b, and 5a) and aromatic (3j-n, 4c, 5b) compounds were prepared, fourteen of them for the first time, and tested for antithrombotic (p.o. administration to rats, 60 mg/kg) effects. The N-(1-cyanocyclohexyl)-N-hydroxy-N'-phenylurea (3j) was most potent and inhibited laser-induced (35 mW, 50 ms) thrombus formation in arterioles by 21% and that in venules by 15%. The compounds form nitric oxide in vitro by the addition of a Fe3(+)-porphyrin complex and an oxygen donor. Moreover, the most active compound 3j in vivo exhibits the highest NO formation in vitro. Furthermore, it was shown that the cyano group is essential for the desired activities and NO formation. These results suggest that the title compounds act as NO donors. PMID- 10675985 TI - Anti-inflammatory and analgesic amides: new developments. AB - A series of substituted N-cycloalkyl benzamides, cinnamamides, and indole-3 carboxamides were synthesized and evaluated for their analgesic, antiinflammatory activities as well as for their gastrointestinal irritation liability. Indomethacin was used as reference drug in both tests. Compounds 1k, 1b, 1h, 1j, and 1g were the most active in the antiinflammatory paw edema inhibition test, with a sharply dose-dependent effect. In terms of the analgesic activity (acetic acid writhing test), the most active compound was 5a followed by 3a, but many other compounds were found to have a non-negligible potency. Even in this case, the effect was dose dependent. PMID- 10675987 TI - First bioanalytical evaluation of nonpeptidic cage dimeric HIV-1 protease inhibitor N-benzyl 4-aryl-1,4-dihydropyridine H17: biotransformation and toxicity on Hep G2 cells. AB - Cage dimeric N-benzyl 4-aryl-1,4-dihydropyridine H17 is a moderate inhibitor of HIV-1 protease. As representative of an innovative and promising class of nonpeptidic HIV-1 protease inhibitors H17 was selected for the characterization of the biochemical profile of the cage dimers concerning metabolic and toxic aspects. In the first bioanalytical evaluation of H17 on Hep G2 monolayers no phase-I metabolites were found and the extent of conjugation on phase-II of biotransformation was poor due to steric hindrance of the hydroxymethylene groups. H17 was found to be nearly non-toxic. A slight noticeable influence on cell proliferation, however, did not result from apoptotic activities. Thus, first biochemical evaluation of H17 practically suggests no decrease of an in vivo bioavailability by metabolization. PMID- 10675988 TI - [Effect of knee joint flexion and femur rotation on retropatellar contact of the human knee joint]. AB - The aim of the present study was to evaluate retropatellar contact characteristics at different angles of flexion of the knee joint. To this end, 6 cadaveric legs were examined using pressure sensitive film (Fuji Prescale type "super low") at angles of flexion of 45 degrees, 60 degrees, 90 degrees and 120 degrees both in neutral rotation and 10 degrees internal and external rotation of the femur in the same knee joints. A force of 140 N was applied to both the vastus medialis and lateralis, and a comparison made with a medially and a laterally dominating muscle force. The contact areas decreased with increasing angles of flexion. The medially dominating muscle traction increased the contact area. Comparison between internal and external rotation revealed a decrease in contact area on internal rotation. The pressure measurements were comparable in all loading situations. Comparison between neutral and medial traction revealed significant differences in contact area, pressure and force. The influence of femoral rotation showed no significant difference. A comparison of the different angles of flexion revealed only few significant differences. To prevent the development of retropatellar arthrosis, maximum contact areas are necessary. The study has shown an advantage for medially dominating muscle traction, and external rotation of the femur. PMID- 10675989 TI - [Measures for biomechanically correct positioning of knee joint endoprostheses]. AB - Normal post-operative function, permanent stable fixation and long life for the implant material are determined mainly by the correct biomechanical positioning of the knee joint prosthesis. We have identified six measures within the surgical procedure that are of vital importance for achieving biomechanically correct positioning. Because of their great importance, we call these measures "The Golden Six", and describe their application using the ESKA Resurfacing Knee System (RKS). This system, which is so designed that it enables a maximal bone saving resection technique, makes use of a diagonal screw-fixation of the endoprosthetic elements, resulting in increased primary stability. For the safe and reliable implementation of our measures, we have developed a special set of instruments. Anatomical, kinematic and surgical studies have shown that with the aid of the above-mentioned six measures biomechanically correct positioning of all the implanted elements can be achieved, thus ensuring trouble-free post operative function of the knee joint and the least possible mechanical stressing of the implanted materials. PMID- 10675990 TI - [Ceramic acetabular cups for hip endoprostheses. 7: How do position of the center of rotation and the CCD angle of the shaft modify range of motion and impingement?]. AB - The range of motion (ROM) of total hip prostheses is influenced by a number of parameters. An insufficient ROM may cause impingement, which may result in subluxation, dislocation or material failure of the prostheses. In a three dimensional CAD simulation, the position of the centre of rotation and the CCD angle of the stem were investigated. Displacement of the centre of rotation of the femoral head may be due to wear (PE cups) or to the design of the prosthesis (ceramic cups). Stems of widely differing design have been developed and implanted. The results of the present study demonstrate that the ROM is clearly reduced by increasing penetration of the femoral head. At an inclination angle of 45 degrees, a depth of penetration of 2 mm restricts flexion by about 15 degrees, and a depth of penetration of 3 mm by about 30 degrees. At smaller angles of inclination the ROM is reduced and flexion and abduction are associated with an increased risk of impingement. With steeper acetabular cup inclinations, the risk of impingement decreases, but dislocation, the risk of rim fractures (ceramic cups), and wear and penetration rates (PE cups) increase. The CCD angle of the stem should be oriented to the anatomical situation. At high CCD angles (> 135 degrees), flexion is clearly limited, in particular when there is penetration of the femoral head. For modern total hip arthroplasty, prosthetic systems characterised by precise positioning of components, minimum wear, slightly recessed inserts, and appropriate CCD angles should be used. PMID- 10675991 TI - [Primary stability of press-fit acetabulum cups using a new acetabulum reamer]. AB - The aim of the present study was to assess the initial stability of uncemented press-fit acetabular components using a newly developed reamer designed to optimize the surgical preparation of the acetabulum. Ten synthetic human pelves were used to investigate the stability of 20 uncemented press-fit acetabular components, each of which was tested in a servohydraulic testing machine for 6 cycles under an axial load of 2.4 kN. The results of the micrometric measurements revealed satisfactory stability for a reaming depth of 2 mm, and a press-fit of 2 mm. Micromotion was less than 200 microns in all the anatomical sections of the acetabulum (ischium 63 microns, pubis 150 microns, ilium 85 microns). A press-fit of 4 mm and the smaller reaming depth of 1 mm were associated with a substantial decrease in mechanical stability. PMID- 10675992 TI - [Primary stability of cement-free press-fit acetabulum cups. In vitro displacement studies]. AB - The initial stability of 6 different hemispherical press-fit titanium acetabular cups was investigated by lever-out tests using a foam model. Each cup was implanted in a standardised manner into machined PVC-foam blocks with an underreaming of 1/2/3 mm, and levered out 5 times. A computer recorded the load displacement curves. Both insertion forces and lever-out forces increased with increasing press-fit from 1 to 2 mm. With underreaming of 2 mm, insertion forces varied between 9 and 34 Nm, and lever-out torques between 13 and 23 Nm. The cup made up of multi-layer irregular titanium wire mesh had the highest initial stability of all the cups tested. 3 mm underreaming was associated with insertion difficulties, so that lever-out is difficult to compare with that with 1 and 2 mm. The amount of contact on the equatorial rim, where we saw the most attrition after lever-out, is the most important factor for determining the lever-out torque. The quality of the PVC foam blocks, the accuracy of reaming, and the definition of cup insertion are very important parameters for test reproduction. Since the trials were conducted under laboratory conditions, translation of the results to the intraoperative situation should be undertaken only with certain reservations. PMID- 10675993 TI - Management of rape survivors. PMID- 10675994 TI - Heat stroke in young adults. PMID- 10675995 TI - Prevalence of Helicobacter pylori infection in patients with functional dyspepsia. AB - OBJECTIVE: To investigate the prevalence of Helicobacter pylori infection in patients with functional dyspepsia. DESIGN: Prospective descriptive study. SETTING: Surgical unit, Base Hospital, Panadura. PATIENTS: 67 patients with persistent symptoms suggestive of functional dyspepsia but no clinical, endoscopic or ultrasonographic evidence of peptic, biliary, pancreatic or malignant disease. METHODS: Upper gastrointestinal endoscopy and two antral mucosal biopsies were performed in all patients and the rapid urease test (CLO test) was done to detect H pylori status. All patients gave informed consent. RESULTS: Of the 67 patients (32 males) with a mean age of 45 years (35 to 70 years), only 2 patients (2.9%) were found to be positive for H pylori infection. CONCLUSION: Incidence of H pylori infection was found to be very low in patients with functional dyspepsia. PMID- 10675996 TI - Hepatitis B and C virus markers among new entrant medical students. AB - AIM: To investigate the presence of hepatitis B and C virus markers in new entrant medical students at the Faculty of Medicine, University of Kelaniya. METHOD: 456 students (mean age 24 years, SD 3.5, 257 men) were investigated before they were exposed to clinical work, using a questionnaire to assess sociodemographic factors and possible risk factors for contracting hepatitis B or C. Blood samples were tested for HBs Ag and anti HBs (n = 456), and anti-HCV (n = 162 randomly selected samples) with a third generation sandwich radioimmunoassay technique. RESULTS: The students were from 20 of the 25 districts in the country, although their distribution was not inform. A past history of hepatitis or jaundice was obtained from 24 (5.3%) and 6 (1.3%) students respectively. None of them had been vaccinated against hepatitis B. At least one risk factor for hepatitis B or C was present in 32 (7%) of them. None of the samples were positive for HBsAg or anti-HCV, and only two (0.44%) were positive for anti-HBs. CONCLUSION: Our results support the view that exposure to hepatitis B and C seems to be uncommon in this country, at least up to young adulthood. As most new entrant medical students are not immune to these infections there is a strong case to vaccinate them against hepatitis B before they are exposed to clinical work. PMID- 10675997 TI - Breast cancer: do histopathology reports provide adequate information for patient care? AB - INTRODUCTION: Many studies have shown that certain pathologic features correlate with risk of recurrence, development of nodal and distant metastases, long term survival and multicentricity of breast carcinoma. Such information is useful in selection of appropriate therapy and prognostication. OBJECTIVE: To determine the proportion of histopathology reports of breast carcinoma that provide essential pathologic prognostic information. METHOD: All (110 mastectomy and 25 lumpectomy) cases of breast carcinoma reported during 79-month period from January 1992 were assessed for inclusion of tumour size, information on excision margins and 8 microscopic features. Mastectomy reports were evaluated for information on axillary lymph node status. RESULTS: 9.6% of reports assessed included all relevant features. All 110 mastectomy reports gave information on axillary lymph node status. Tumour size was included in 91.1% of reports. Excision margins were commented on in 36% of lumpectomy reports. Of the microscopic features assessed, tumour type was included in 98.5% and histologic grade in 66.7%. In formation on other features ranged from 28.1% for inflammatory infiltrate at the tumour-host interface to 69.1% for nipple invasion. CONCLUSION: Reporting on basic prognostic information falls short of the ideal. Provision of guidelines will help to ensure the inclusion of relevant histopathologic prognostic information. PMID- 10675998 TI - Do babies need water in Sri Lanka? AB - OBJECTIVES: To study the prevalence of exclusive breast feeding and the reasons for water supplementation, and investigate whether water is necessary in the humid climate of Colombo. SETTING: Well baby clinic in De Soysa Maternity Hospital for Women, Colombo. SUBJECTS: 200 breast fed infants born in a baby friendly hospital between the ages of 1 and 4 months. METHODS: The study sample was randomly selected. Sick infants and those of working mothers were excluded. The mothers' knowledge of feeding practices was recorded in an interviewer administered questionnaire. The infants' weights, lengths and rectal temperatures were measured and the osmolality of urine estimated. The room temperature and relative humidity were recorded on each day of study. The data were analysed using the Chi-Square statistical test. RESULTS: 69% of mothers introduced supplementary fluids within the first 4 months, because of advice from grandmothers or relatives, thirst, hiccups or constipation. 45% had introduced water with "rathakalkaya", a fluid traditionally given to infants in Sri Lanka. 90% of mothers had attended antenatal clinics in the De Soysa Hospital for Women. 70% of mothers who gave supplementary fluids were aware of the importance of exclusive breast feeding. The range of urine osomolalities of exclusively breast fed infants was 60 to 204 mosmol/kg. There were no significant differences in core temperatures and number of times urine was voided daily, or urine osmolalities, between exclusively breast fed and fluids-supplemented group of infants. CONCLUSIONS: Despite delivery in a baby friendly hospital a majority of mothers supplemented breast milk with water or other fluids during the first 4 months. The advice of grandmothers had a significant influence on early feeding practices. Exclusively breast fed infants were found to maintain water homeostasis under the hot, humid climatic conditions of this study. PMID- 10675999 TI - A family with alkaptonuria showing quasidominant inheritance. PMID- 10676000 TI - Right ventricular arrhythmogenic dysplasia in a young Sri Lankan. PMID- 10676001 TI - Tilt table test and assessment of neuro-cardiogenic syncope. PMID- 10676002 TI - Micropapillary serous carcinoma of the ovary: a report of three cases. PMID- 10676004 TI - Emergency contraception. PMID- 10676003 TI - Emergency contraception. PMID- 10676005 TI - A preliminary study on neonatal septicaemia in a tertiary referral hospital paediatric unit. PMID- 10676006 TI - Vitamin D supplements in adults. PMID- 10676007 TI - Anti-ulcer drugs may mask early gastric cancer. PMID- 10676008 TI - [Complications after joint endoprosthesis for treatment of locomotor system neoplasms]. AB - The authors present most common complications after tumor related joint replacement. Twenty-six patients (9 females, 17 males) have been operated on (mean age 22.5 years). In 19 cases osteosarcoma was diagnosed, Ewing sarcoma in 4 and there were 3 cases of chondrosarcoma. Postoperatively, superficial skin necrosis occurred in 4 patients but healed in four weeks, full-thickness skin necrosis in 2 and in 2 cases wound infection resulted in septic loosening of endoprosthesis. The number of infections in our patients was low but caused delay of chemotherapy and need for revision of the infected joint. PMID- 10676009 TI - [Early results of densitometry around the cementless stem of the Parhofer-Monch hip endoprosthesis]. AB - Fifty-three patients with unilateral hip arthritis (31 females, 22 males) aged from 29 to 66 years (mean 52.1 years) after total hip replacement with cementless Parhofer-Monch endoprosthesis underwent densitometry. The content (BMC) and mineral density (BMD) of calcium within proximal end of the femur was evaluated at 8-12 days postoperatively and at 6, 12 and 18 months follow-up. Early results show various mineral changes in Gruen zones around the stem. The greatest BMC and BMD decrease was found at 6 months follow-up in zones 1 and 7 as well as 2 and 6. The BMC and BMD tendency to decrease gradually ceased and slightly positive values were found in zone 3, 4 and 5 at the final evaluation. PMID- 10676010 TI - [Using arthroscopy for dynamic assessment of the patello-femoral joint in patients with anterior knee pain]. AB - Arthroscopic assessment of patello-femoral dynamics in 20 females aged from 15 to 23 years with anterior knee pain syndrome has been compared to MRI study done in this group previously. Arthroscopic evaluation was done according to Lindberg et al. The study suggests, that arthroscopy is an useful technique in evaluation of patello-femoral instability, giving a chance for treatment of other intraarticular disorders (if present) contributing to knee pain. PMID- 10676011 TI - [Free vascularized joint transfer during surgery for hypoplastic thumb]. AB - Hypoplasia and aplasia of the thumb constitute approximately 11% of congenital disorders of the upper extremity. Surgical treatment depends on type of disorder in Blauth classification used in our Department. Soft tissue procedures and pollicisations are performed most often. Microsurgical transplantation of interphalangeal joint from the toe is an alternative. This paper describes a case of microsurgical treatment in 20 years old patient with hypoplastic thumb. Two months after surgery the range of motion in transplanted joint was between 20 and 75 degrees. PMID- 10676012 TI - [Does delayed fracture fixation within the upper extremity influence the rate of complications?]. AB - A series of 42 patients operated on due to humeral shaft fracture and 67 patients with forearm bones shaft fracture were analyzed clinically and statistically. The type of fracture was related to the time span between injury and surgery and number of complications. Complications were divided into two groups--early ones (skin necrosis, infection) and late ones (fixation failure, delayed union, pseudoarthrosis). Injury to surgery time span was divided into for sections: less than 10 h, 10-24 h, 24 h-10 d and above 10 days. Early and late complications prevailed in patients operated within 24 hours from injury (87% of all complications). Eleven patients with humeral fracture had fair or poor result; 8 of them were operated within 24 hours from injury. Among 18 patients with fair and poor results after forearm fracture 13 were operated on within 24 hours from injury. PMID- 10676013 TI - [Percutaneous epiphyseodesis for treating limb length inequality in children - results after termination of growth]. AB - Results of percutaneous epiphyseodesis for limb length egalization in 25 children (10 girls and 15 boys) after termination of growth are presented. Mean age at the surgery was 12.9 years (range 10.9-14.4 years), mean limb length discrepancy 4.3 cm (range 2-10 cm), mean follow-up was 50 months (range 14-86 months). In 17 cases distal femoral epiphyseodesis was done, in 7 cases distal femoral, proximal tibial and fbular and in 1 case only proximal tibial and fibular epiphyseodesis was performed. In 13 cases CT served to assess the area of removed physis. Among complications knee hemarthrosis was encountered twice and once subcutaneous hematoma of the lower leg occurred. At the final follow-up full range of motion in the knee and correct axial alignment was observed in all cases. Residual limb length discrepancy was 1.3 cm (range 0-4 cm). Between 20 and 60% of total area of the physis was destroyed. Yearly rate of femoral bone growth inhibition was the same in patients with 27% and 40% of physis removed. PMID- 10676014 TI - [Ultrasonographic evaluation of the rotator cuff after its surgical reconstruction]. AB - The aim of this paper was to evaluate ultrasonographic picture of rotator cuff integrity after surgical reconstruction performed in 34 patients with mean follow up 17.2 months (range 6-45 months). The lack of rotator cuff echo or hypoechogenic area were the criteria for diagnosis of re-tear. This occurred in 16% of shoulders with isolated supraspinatus tear and in 33.3% of shoulder with 2 tendons involved. Hyperechogenic changes were noted in 35.3% of shoulders, some thinning of the rotator cuff in 23.5%, while both of these changes in 14.7%. The thinning of the rotator cuff was classified as pathologic if there was difference between diameter of rotator cuff measured in 6th to 8th postoperative week and diameter measured at the time of follow-up. Ultrasonographic examination offers critical evaluation of both surgical result and postoperative physical therapy protocol. PMID- 10676015 TI - [Radiologic changes in hip joints of patients with hemophilia]. AB - Historical overview and contemporary understanding of hemophilia and hemophilic arthropaty is presented. Avascular necrosis of the femoral head occurs in 7% of hemophilic patients. Clinical description and hip radiographs of 6 patients were presented and discussed according to Nuss modification of Arnold-Hilgartner classification. PMID- 10676016 TI - [Cross-leg pedicled fibular graft--a new reconstructive method with vascularized bone graft]. AB - Theoretical foundations for cross-leg pedicled fibular graft, a new method for reconstruction within lower extremity with vascularized bone graft is presented. The flap can be raised in two fashions depending on blood flow direction in the pedicle. In case of regular flow up to 18 cm of fibula is available; in reversed pedicular flow over 20 cm of fibula can be harvested. An arch of rotation of the flap reaches mid-thigh and peripheral part of the lower extremity. Cross-leg pedicled fibular graft can be used for concomitant soft tissue defect reconstruction as well. Crossing and immobilization of both lower extremities is necessary for 4 weeks. No microsurgical procedure is required. Skin island of the fibular flap or narrow muscular cuff left around fibular vessels is sufficient to protect the pedicle. Main indication for cross-leg pedicled fibular flap include patients with major lower extremity injury with axial vessels damage or with history of previous trauma and thrombosis, and patients after bone tumor resection who had chemotherapy and/or radiotherapy. The indications may be markedly broadened especially in centers with no access to microsurgery. The results of this method are very encouraging so far. PMID- 10676017 TI - [A modified direct lateral approach for total hip arthroplasty]. AB - Literature based review of lateral hip approach (MacFarland, Osborne) modifications offered by Bauer, Harding, Mills, Moscal and Mann is presented. Surgical technique for the Moscal and Mann approach is described. The approach allows easy access to the proximal femur for reaming, cementing, prosthesis orientation and leg-length discrepancy correction. It offers visualization of the joint superior to all non transtrochanteric approaches and is successfully used for both primary and revision THR. PMID- 10676018 TI - [Clinical use of underoot pressure measurements done with the Emed-system for diagnosis of foot deformities and disorders]. AB - Emed-system is used for assessment of pressure distribution under the foot at standing and walking since 1985. The method is non-invasive, useful in diagnosing and monitoring of treatment for foot disorders both in children and adults. It may be utilized in orthopedics, traumatology, rehabilitation, rheumathology and diabetology. PMID- 10676019 TI - [A method for gait analysis--preliminary report]. AB - A preliminary report on new method for gait evaluation is presented. Specifically designed shoes with built-in sensors allow for registration of the data during free gait on normal surface; collected data is than analyzed in PC based system. Twenty-four patients (16 females, 8 males, mean age 58 years) with hip arthritis treated at Orthopedic Department were included in this study. PMID- 10676020 TI - [Covering soft tissue defects after tumor resection around the knee with vascularized musculo-cutaneous and fascio-cutaneous flaps]. AB - The application of musculo-cutaneous of the medial head of gastrocnemius for covering skin and soft tissues defects following total knee replacement in treatment for proximal tibial osteosarcoma in 4 patients is presented. In 2 patients the fascio-cutaneous flaps originated from lower leg were used to cover defects of anterior aspect of the knee after resection of dermatofibrosarcoma of the patellae and fibrohistiocytoma localized in proximal tibia. PMID- 10676021 TI - Amputations and prosthetics. AB - The author presents in a condensed way an overview of the principles of limb amputations and further treatment of patients who underwent such a procedure. The metabolic cost of walking, load transfer, and wound healing are reviewed in a concise manner. Particular attention is given to blood supply to the wound and methods to determine adequate perfusion with a clear analysis of the pro and cons of the Doppler method. Pediatric amputations, because of their specificity, are considered apart. Disarticulation of limbs is the method of choice in children, because of it retains growth potential of the bone and prevents bony overgrowth of the stump. The article discusses the main indications for limb amputations: trauma, peripheral vascular disease, musculoskeletal tumors and gas gangrene. In every case the specificity of the amputation is considered by the author. Postoperative care is also presented, with a short description of possible complications. Pain is the most common and treatment strategies should be similar to those used in treating patients with major reflex sympathetic causalgia. Edema, joint contracture, wound failure and dermatologic problems are all shortly reviewed. The last part of the article treats with the principles of prosthetics in both the upper and lower limb. These principles are presented basing on the level of amputation: for the upper limb hand, transradial, transhumeral amputations and shoulder disarticulation. For the lower limb foot and ankle, transtibial and transfemoral amputations are considered. PMID- 10676022 TI - [Ruptured hepatocarcinoma. Report of 22 cases]. AB - STUDY AIM: Spontaneous rupture of hepatocellular carcinoma (HCC) causing massive hemoperitoneum is a critical and life threatening complication. The study aim was to report a retrospective series of 22 cases observed in the same centre. PATIENTS AND METHODS: From 1978 to 1998, 22 patients (18 males and four females, mean age: 63 years, range: 18-83) were treated for ruptured H.C.C involving a cirrhotic liver in 17 cases and a normal liver in five cases. In 14 cases, the diagnosis of acute hemoperitoneum indicated an immediate laparotomy. The site of rupture was predominant in the left lobe (eight cases). The surgical treatment was: left lobectomy (n = 7), right hepatectomy (n = 2), excision (n = 4), hepatic artery ligation (n = 5), direct hemostasis (n = 4). RESULTS: Postoperative mortality was 45.4%. Among the 12 survivors, nine died within a delay of 6 to 29 months. Three patients were still alive at the time of this study at 32, 40 and 66 months. CONCLUSION: Acute rupture of HCC requires emergency procedures with a high risk of mortality. Curative operation with hepatic resection is the most effective procedure but is not often feasible because of the spreading of the tumor or/and the cirrhosis. The ligation of hepatic artery seems to be an alternative procedure to obtain an immediate hemostasis. Fissuration allows performance of complementary explorations and possibly preoperative arterial embolization with better immediate results. PMID- 10676023 TI - [Hepatectomy in intrahepatic lithiasis]. AB - STUDY AIM: The aim of this study was to report the immediate results of a series of 65 hepatic resections for hepatolithiasis performed in Vietnam. PATIENTS AND METHOD: From 1986 to 1998, 44 men and 21 women (mean age: 40 years) underwent hepatic resection for hepatolithiasis. Fourty patients had previously undergone one or several operations for hepatolithiasis. The procedure was performed on emergency in 25 patients. Indications for hepatic resection were: angiocholitis and liver abscess in 22 cases, stones closely inserted in the biliary duct in 20 cases, hemobilia in 12 cases, stones located above a biliary stricture in 8 cases and stones associated with a postoperative biliary fistula in 3 cases. Liver resections (minor in 61 patients, including 55 left lobectomies, and major in 4 patients) were performed through transhepatic approach according to the Ton That Tung technique and followed by an external biliary drainage with a Kehr tube. RESULTS: There were 6 postoperative deaths (9%), 3 due to septic shock, 2 to cachexia, and 1 to liver failure. The 15 patients with complications recovered with conservative therapy. Bile infection was present in 93%, mostly with Escherichia coli and Enterobacter. Pigmented stones were usually found. CONCLUSION: Vietnam is a country with high incidence of hepatolithlasis. Hepatic resection is an adequate treatment for localized intrahepatic bile duct stones when the involved segment including biliary strictures and calculi can be completely removed. The procedure may be performed on emergency for liver abscess, or hemobilia. PMID- 10676024 TI - [Selective use of vascular clamps in major hepatectomy]. AB - OBJECTIVE: To report the results of a selective use of vascular occlusions in major hepatectomies according to the size and location of the hepatic lesion. BACKGROUND: Total vascular exclusion (TVE) and portal triad clamping (PTC) ensure efficient hemostatic effect but lead to warm ischemia of the liver. Lobar vascular occlusion (LVO) avoids warm ischemia of the remnant liver but could result in increased blood loss. PATIENTS AND METHODS: Sixty consecutive major hepatectomies were studied. TVE was applied in 22 patients with large lesions (= 10 cm) or lesions with connections to the major hepatic veins or inferior vena cava. PTC (n = 15) and LVO (n = 23) were applied in remaining cases. RESULTS: Clamping method was efficient in 87%, 93% and 100% for LVO, PTC and TVE, respectively. Median blood transfusions were 0.3 and 2 units for LVO, PTC and TVE, respectively. Postoperative aminotransferase peak value was significantly lower after LVO than after PTC or TVE, while those peaks were not statistically different with these latter two methods. Postoperative prothrombin time fall value was identical in the three groups. Mortality was 3.3% (2/60) and was not influenced by the type of clamping, but both deaths and most complications occurred in patients with abnormal underlying liver parenchyma. CONCLUSION: Provided that adequate techniques are used, the need for blood transfusions is more dependent on the characteristics of the resected tumor than on the type of clamping used. Total vascular exclusion does not create more ischemic injury to the liver than portal triad clamping and it should be recommended for the resection of large or strategically located tumors. Other tumors can be resected in more than 80% of the cases with LVO, thus avoiding ischemia to the remnant liver. With the control of hemorrhage, pathology of underlying liver parenchyma has emerged as the main prognostic factor in major liver resections. PMID- 10676025 TI - [Adjuvant intra-arterial chemotherapy after curative resection of liver metastasis from colorectal cancer. Results of a pilot study in 30 patients]. AB - OBJECTIVE: Five-year survival after simple resection of liver metastases from colorectal carcinoma ranges from 20 to 40%. The aim was to study the reliability and long term results of adjuvant intra-arterial chemotherapy after resection of colorectal liver metastases. PATIENTS AND METHOD: From 1991 to 1997, 30 patients after a complete resection of liver metastases from colorectal cancer were included (16 men, 14 women, mean age: 62 years). There were 2 stage I, 19 stages II, 2 stages III, 5 stages IV and 2 stages V according to Gayowski staging system. During laparotomy, a catheter was placed in the gastroduodenal artery in order to perfuse the proper hepatic artery. Chemotherapy included 5 Fluorouracil (12 mg/m2) and Leucovorin (200 mg/m2) and was administered once a week during six months. Mean follow-up was 52 months. RESULTS: Adjuvant intra-arterial chemotherapy had to be interrupted before six months in 9 patients because leukopenia (n = 2), infection or obstruction of the catheter (n = 5), duodenal migration of the catheter (n = 1) and occurrence of multiple extrahepatic metastases (n = 1). No death was in relation with the method. Five-year survival rate was 41.8% for the global series. Five-year disease free survival rate was 21.4%. Causes of death were: hepatic recurrence only (n = 3), extrahepatic + hepatic recurrence (n = 4), extrahepatic recurrence (n = 2). Two patients died of another carcinoma (esophagus, ovary), without evidence of recurrence of the colorectal carcinoma. At the present, there is a recurrence in 4 living patients. CONCLUSION: Although the benefit on survival is not significant, these results suggest a longest time of remission in patients with adjuvant intra-arterial chemotherapy. Trials comparing and/or combining this method to intravenous chemotherapy should be proposed in patients after resection of colorectal liver metastases. PMID- 10676026 TI - [Mycotic aneurysm after kidney transplantation]. AB - PURPOSE: The study aim was to report six cases of mycotic aneurysms in renal transplant patients and to review the literature on this subject. PATIENTS AND METHODS: Six patients, aged from 13 to 59 years, who had undergone renal transplantation 4 months to 16 years earlier, developed a mycotic aneurysm after bacteremia. The diagnosis was based on morphological investigations (echotomography, arteriography, spiral computed tomography) and bacteriological studies (blood culture, culture of the aneurysmal wall and content). The aneurysm was located in five cases at the anastomosis of the renal artery with the iliac axis, and in one case on the popliteal artery and tibioperoneal trunk. All patients were treated surgically: five reconstructions were performed using two arterial iliac prostheses, three hypogastric artery autografts and one saphenous vein graft (combined with an iliac prosthesis); one repair was impossible because of profuse local suppuration, and endoaneurysmorraphy with multiple ligatures of the popliteal vessels was performed. Postoperative radiological control was performed in all cases of arterial repair. All patients received antibiotic therapy during three to six months after the operation. RESULTS: No postoperative mortality occurred. All kidney transplants were salvaged. Anatomical results of arterial reconstructions were satisfactory in all cases and remained so during the follow-up. CONCLUSIONS: Mycotic aneurysms after renal transplantation are rare since only six observations with a kidney transplant in place have been published in the literature with a single long-lasting kidney salvage. Surgical treatment is mandatory to prevent rupture. Survival of patients occurred exclusively in operated cases. PMID- 10676027 TI - [Pulmonary aspergilloma: results of surgical treatment. Report of a series of 206 cases]. AB - STUDY AIM: The aim of this retrospective study was to report the results of the surgical treatment in 188 patients operated on for pulmonary aspergilloma in a series of 206 patients observed in Morocco. PATIENTS AND METHOD: From 1982 to 1998, 206 patients were treated for pulmonary aspergilloma in the same hospital; 188 were operated on and surgery was contraindicated in the other patients with general or respiratory failure. Hemoptysis was the main symptom, present in 190 patients (92%). Surgery was performed on principle with 108 lobectomies, 38 segmentectomies, 18 lobectomies and segmentectomies, 21 pleuropneumonectomies and 3 thoracoplasties. RESULTS: Postoperative complications occurred in 36% of the patients including: pyothorax (n = 15), hemothorax (n = 10), rehabitation defects (n = 17) and respiratory failure (n = 10). Reoperation was necessary in 6 patients. Postoperative mortality rate was 6.4% (12 patients including 5 treated by pleuro-pneumonectomy). CONCLUSION: The surgical treatment, in spite of its high morbidity, has to be proposed to all patients with pulmonary aspergilloma, even in asymptomatic patients when there is no surgical contraindication. Pleuropneumonectomy is a very high risk procedure and its indications must be restricted. Thoracoscopy was rarely performed in this series. PMID- 10676028 TI - [Use of lanreotide in the prevention of pancreatic fistula after cephalic duodeno pancreatectomy. Preliminary study]. AB - STUDY AIM: Dehiscence of pancreatic anastomosis is the main complication after pancreatoduodenectomy. The efficacy of somatostatin analogue to prevent complications after pancreatic resections is at present well-established by several randomized trials. The aim of this preliminary prospective study was to assess the role of lanreotide (a long acting somatostatin analogue) in this field. PATIENTS AND METHOD: Forty patients with pancreatic head tumour have been included in a prospective study. Criteria for pancreatic fistula were: high concentration of amylase in the drainage fluid (> 3 times that in the serum), or intra-abdominal fluid collection adjacent to the pancreatic anastomosis, or reoperation (or postmortem verification) showing an anastomotic dehiscence. The patients received 12 h before the operation 30 mg of lanreotide intramuscularly. RESULTS: Of the 40 patients included prospectively, 34 underwent a pancreatic resection. Parenchyma of pancreatic remnant was crumbly in 28 cases. Six patients experienced a pancreatic fistula (17.6%) which healed in all cases. CONCLUSION: This preliminary study shows clearly the feasibility of a long acting somatostatin analogue (lanreotide) to prevent pancreatic fistula after pancreatectomy. This agent appears simple to use and its efficacy needs obviously to be assessed by randomized trials. PMID- 10676029 TI - [Treatment of hemorrhoids with the Longo technique. Preliminary results of a prospective study on 94 cases]. AB - AIM OF STUDY: The aim of this multicenter prospective study was to report the early results of Longo procedure for the surgical treatment of hemorrhoids disease. PATIENTS AND METHOD: From April 1998 to July 1998, 94 patients (60 men and 34 women with a mean age of 47 years) were treated according to Longo procedure for a mucosal prolapse (12 grade II, 63 grade III, and 19 grade IV). All patients were evaluated at 2 and 6 postoperative months. The technique consisted in the reduction of mucosal and hemorrhoidal prolapses with a circular suturing device. RESULTS: Postoperative morbidity rate was 6.3% (n = 6). A rectal bleeding occurred within 12 hours after surgery in five patients. The mean postoperative length of hospital stay was 36 hours (range: 24-72 hours). The only antalgic prescribed was paracetamol. Local care was not necessary in any patient. After 6 months, 89 patients (94.7%) were very satisfied, three patients (3.2%) were satisfied (rectal sub-mucosal abscess in one case, functional troubles in two cases) and two patients (2.1%) were not satisfied (persistence of mucosal prolapse). CONCLUSION: These preliminary results are satisfactory but need to be confirmed by a prospective randomized trial, comparing Milligan Morgan procedure and Longo procedure. PMID- 10676030 TI - [Surgical treatment of colonic cancer after 75 years of age. Study of a series of 240 patients]. AB - AIM OF THE STUDY: The aim of this retrospective study was to report the results of the surgical treatment for large bowel cancer in patients over 75 years of age. PATIENTS AND METHOD: From 1985 to 1996, 240 patients. 114 men and 126 women, aged 75 years or over (mean age: 82, range: 75 to 95 years) underwent surgical treatment for large bowel carcinoma. Tumors were located in the right colon (n = 120), left colon (n = 100), transverse colon (n = 5), or were multiple (n = 15). Clinical presentation was failure of general condition (25%), intestinal obstruction (20%), rectal bleeding (20%), abdominal pain (17%). ASA score was I (n = 1), II (n = 69), III (n = 134), or IV (n = 36). Emergency surgery was mandatory in 110 cases (43 urgent and 67 delayed procedures) and 130 patients underwent elective surgery. Surgical resection was performed in 221 cases, including 177 cases with curative intent (67%). Surgical procedures included right colectomy (n = 119), left colectomy (n = 59), transverse colectomy (n = 9) or subtotal colectomy (n = 31). Histopathological staging was Astler--Coller A (n = 8), B (n = 116), C (n = 54) et D (n = 62). RESULTS: The postoperative course was uneventful in 157 cases (65.4%). Medical complications occurred in 46 patients with 34 deaths; and surgical complications in 39 patients with 20 subsequent reoperations and 15 deaths. The overall postoperative mortality rate was 20.4% (n = 49). Postoperative mortality rate was higher after emergency operations (32.7% vs 10%), higher with the level of ASA grading (class II: 8.6%, Class III: 17.1%, Class IV: 38.8%), higher in patients over 90 years (37.4% vs. 19.1%) and in patients without surgical resection (42% vs 18.5%). Disease specific 5-year survival rate was 45% and did not differ when compared to patients younger than 75 years (42%, data not reported). CONCLUSIONS: Patients older than 75 years remain a high risk group, specially if operated on emergency. Nevertheless, age is not a limiting factor in the surgical treatment of colon cancer. Prognosis is mostly depending on ASA grading. Colectomy with curative intent has to be performed when possible. PMID- 10676032 TI - [The first dissecting room for surgeons in Paris]. PMID- 10676031 TI - [Impact of learning and experience on the laparoscopic treatment of gastroesophageal reflux]. AB - STUDY AIM: Laparoscopic treatment of gastroesophageal reflux disease (GERD) by partial (PF) or total (TF) fundoplication is the most appropriate surgical treatment after failure of medical treatment. The aim of this study was to compare the results of the same series in three consecutive periods in order to determine the effects of the learning curve and experience on the technique and outcome. PATIENTS AND METHODS: From January 1993 to January 1998, 150 patients (84 men and 66 women) with a mean age of 52.2 years (18 to 78) were included. Three groups of 49, 50 and 51 patients were chronologically defined. The comparison was established on the following criteria: the operative technique; the conversion rate; the mortality and morbidity rate; the duration of surgery and hospitalization and the results with short and medium follow-up. RESULTS: The three groups were comparable with respect to patients and GERD characteristics. One hundred and thirty two patients had a TF and 18 had a PF. Rossetti's type TF became the reference procedure (80.3% in group III) and closure of the diaphragmatic crura was performed systematically in group III (100%). The duration of surgery was significantly reduced between group I and the two other groups (138, 100, 80 min). The rate of conversion decreased from 10.2% to 4% and then 0%. The average duration of hospitalization decreased from 5.8 to 4.2 days (p = 0.01). There was no mortality and the morbidity rate decreased from 14.3% to 4% and then 0%. Seven cases of recurrence occurred (4.6%), 5 in group I (10.2%), 2 in group II (4%), and 0 in group III, (with a shorter follow-up). CONCLUSION: The effect of the learning curve has to be taken into account in the training of surgeons (within experienced departments, with "guidance" during initial interventions) and also in the evaluation of results, in order to allow a more accurate comparison between the different treatments for GERD. PMID- 10676033 TI - [Long-term functional results of Nissen fundoplication with laparotomy and laparoscopy]. PMID- 10676034 TI - [Long-term results of biliary repair of laparoscopic bile duct injuries]. PMID- 10676035 TI - [Carotid endarterectomy with patch versus reversion and reimplantation endarterectomy: randomized prospective study]. PMID- 10676037 TI - [Theory and practice of daily prescription and gynecologic consultation for treatment of hyperandrogenism. Indications and contraindications]. PMID- 10676036 TI - [The HPV test and clinical practice]. PMID- 10676038 TI - [What remains of the postcoital test?]. PMID- 10676039 TI - [Are cervicovaginal smears feasible in women over 65 years under hormone replacement therapy?]. PMID- 10676040 TI - [Should cytological screening for cervical cancer be stopped after menopause?]. PMID- 10676042 TI - [Cytological screening of uterine cervical cancer by samples in liquid medium (CytoRich). Preliminary study of a series of 111 292 patients]. AB - We report on the preliminary results of a series of 111,292 patients who benefited from a liquid medium sample (CytoRich) for cervical cancer screening. The number of dubious or limited smears was reduced by 0.03% and 0.53% respectively. The junction zone was better explored, and metaplastic changes were observed in 35.71% of the cases. This method improved the identification of low grade lesions by +56% (2751/111,292; 2.47%) and of high-grade lesions by +75% (860/111,292; 0.77%), with a reduction in the number of ASCUS/AGUS by -44% (2065/111,292; 1.85%). This preliminary study confirms the results already published. The results demonstrating cytohistological correlation should prove to be a decisive factor, enabling the testing of the sensitivity and specificity of this technique. It will then be possible to envisage a future 'new paradigm' for screening cervical cancer as the result of a liquid medium sample, computer assisted screening and HPV viral identification by Hybrid capture II. PMID- 10676041 TI - [Diabetes before pregnancy, apropos of 143 cases]. AB - OBJECTIVE: The authors want to appraise the management of diabetes prior to pregnancy in a local population treated in the Lille University Hospital. METHOD: This is a retrospective study of 143 pregnancies occurring in 111 patients with diabetes prior to pregnancy, between 1987 and 1997, in the Obstetrics Department at the Lille University Hospital. RESULTS: Only one-third of the patients benefited from preconception management; the stability of diabetes during the first trimester was satisfactory in 50% of the cases. The maternal complications are represented by preeclampsia (20%), metabolic complications specific to diabetes (hypoglycemia, ketoacidosis), the aggravation or the emergence of a retinopathy (10%) and polyhydramnios (19%). Concerning the termination of the pregnancies, of the 147 fetuses (four twin pregnancies), 140 newborns in good health, two neonatal deaths, three in-utero deaths and two therapeutic terminations of pregnancy were observed. The fetal malformation rate was 9.5% (14 cases/147). The cesarean section rate was 63%, whereas the fetal macrosomatia rate was 35%, with dystocia in 26% of the deliveries (outside of planned cesareans). Three shoulder dystocia were observed (two requiring the Jacquemier's maneuver and one with transitory plexus brachial palsy for a newborn weighing 5,650 g). CONCLUSION: The authors conclude that preconception management (one third of the patients in this series) and management of during the first trimester of pregnancy (50% in this series) was insufficient. This fact is perhaps due to the confusion, for many practitioners, with gestational diabetes, which is a very mediatized affection, though much less severe for the fetus and mother. PMID- 10676043 TI - [Effects of raloxifene on bone loss and fracture risk in the menopausal woman]. AB - The relevance and efficacy of long-term estrogen therapy is well established, though some undesirable side effects and contraindications persist. Raloxifene, the first selective estrogen receptive modulator (SERM) tested in phase III trials, offers a choice alternative. It increases bone mineral density, lowers serum lipid concentrations and reduces vertebral fractures. PMID- 10676044 TI - [Anatomoclinical correlations of endometriosis]. AB - This is an epidemiological study of 1,498 patients who underwent laparoscopic surgery for different reasons. Between 1989 and 1996, 308 cases of endometriosis were identified among 1,498 patients who underwent laparoscopic surgery. One hundred and five patients were admitted for pelvic pain, 794 for infertility, 319 patients had both on admission, and 280 were admitted for non-gynecologic complaints. The incidence of endometriosis is related to the chief complaint on admission. This disease has different clinical manifestations, different locations and different stages. The mean age in our series is greater than that reported by the literature. The symptoms are related to the location of the lesions but not the stage of the disease. Unlike pelvic pain, infertility is correlated to the stages of the disease. PMID- 10676045 TI - [Proximal tubal unblocking by selective salpingography, apropos of 8 cases]. AB - Proximal tubal desobstuction by selective salpingography: through a study of eight cases, the authors give their first results about tubal desobstruction by selective salpingography. PMID- 10676046 TI - [Inflammatory and/or locally advanced breast cancers. Difficulty evaluating the response to initial chemotherapy]. AB - At the Tenon Hospital (Paris), 48 inflammatory or locally advanced breast cancers were treated by neoadjuvant chemotherapy (three different protocols), followed by surgery (mastectomy or tumorectomy and axillary dissection). The histological data of the specimens are analyzed with regard to the clinical and radiologic evolution. Twenty complete clinical responses (41%), 22 partial clinical responses (45%), and no response in six cases (14%) were observed. Histology demonstrated residual tumors in 42 cases (87.5%), even in the case of complete clinical response (14 cases), without notable change of the grade (SBR). Lymph node clearance demonstrated positive nodes in 38 cases (79%). Despite its great agreement with clinical examination, mammography does not predict the existence of histological residual tumors after initial chemotherapy. Chemotherapy allowed local treatment, but can rarely sterilize the tumor completely. PMID- 10676047 TI - IMS to support Medicare lawsuit. PMID- 10676048 TI - Reporting impaired drivers. PMID- 10676049 TI - Antibiotic resistance: a critical issue in Iowa. PMID- 10676050 TI - OSHA focuses on safe needle devices. PMID- 10676051 TI - Is your office handicap compliant? PMID- 10676052 TI - Don't jump the gun.... PMID- 10676053 TI - [Pediculosis and phthiriasis]. PMID- 10676054 TI - [Viral skin infections in children]. PMID- 10676055 TI - [Skin and sun: prevention and risks]. PMID- 10676056 TI - Determinants of use of maternal-child health services in rural Ghana. AB - This study uses data from the Ghana Demographic and Health Survey (GDHS) of 1993 to examine factors determining the use of maternal-child health (MCH) services in rural Ghana. The MCH services under study are: (1) use of a doctor for prenatal care; (2) soliciting four or more antenatal check-ups; (3) place of delivery; (4) participation in family planning. Bivariate and multivariate techniques are employed in the analyses. The analyses reveal that the use of MCH services tends to be shaped mostly by level of education, religious background and region of residence, and partially by ethnicity and occupation. The implications of these results are discussed. PMID- 10676057 TI - Women's work and fertility in a sub-Saharan urban setting: a social environment approach. AB - Data from three separate studies conducted in Maputo, Mozambique, in 1993 are used to analyse the relationship between the type of social environment in which women work and their fertility and contraceptive use. The analysis finds that women who work in more collectivized environments have fewer children and are more likely to use modern contraception than women who work in more individualized milieus and those who do not work outside the home. Most of these differences persist in multivariate tests. It is argued that collectivized work environments are most conducive to diffusion and legitimation of reproductive innovations. In contrast, individualized environments tend to isolate women and therefore may retard their acceptance of innovative fertility-related behaviour. PMID- 10676058 TI - Hysterectomy is associated with postmenopausal body composition characteristics. AB - The impact of hysterectomy without oophorectomy and with no malignant purpose on body composition and postmenopausal weight gain was tested in 184 Viennese females aged between 47 and 57 years (mean 52.9). Hysterectomized women were significantly heavier than those who experienced a spontaneous menopause (controls). The amount of fat tissue, especially in the abdominal region, was significantly higher in hysterectomized women. Furthermore, they were reported to have experienced a significantly higher weight gain since menopause (9.1 versus 6.0 kg). No significant differences in bone mass were found. Psychological stress factors and hormonal changes following hysterectomy are discussed as possible causes of these differences. PMID- 10676060 TI - The association between health-related behaviours and the risk of divorce in the USA. AB - This study investigates the link between health-related variables and risks of divorce. The findings indicate that physical characteristics associated with poor health--namely, obesity and short stature--are not significantly related to risks of marital dissolution for either men or women. On the other hand, risk-taking behaviours--such as smoking and drug use--are strongly related to higher risks of divorce for both sexes. Overall, the results emphasize the need to accommodate health-related variables in the dominant economic and social psychological theories of marital dissolution. PMID- 10676059 TI - Breast-feeding, diarrhoea and sanitation as components of infant and child health: a study of large scale survey data from Ghana and Nigeria. AB - Using Demographic and Health Survey datasets from Ghana and Nigeria, this study examined whether the protective effects of breast-feeding are greatest where the poorest sanitation conditions prevail. It was found that mixed-fed infants aged between 0 and 11 months tend to have a higher risk of diarrhoea than fully breast fed children, while the risk of diarrhoea among weaned infants is twice that of mixed-fed infants. The probit regression models employed in the analysis were used to predict the probability of diarrhoea associated with each breast-feeding pattern for both 'poor' and 'good' sanitation areas. It was found that the risk of diarrhoea among mixed-fed infants in the poor sanitation areas tends to be high while the same risk among fully breast-fed infants tends to be minimal. In essence, the health risks of mixed feeding are real, particularly for infants aged less than 7 months, and are even worse for those weaned before 6 months of age. PMID- 10676061 TI - The impact of co-resident spouses and sons on elderly mortality in rural Bangladesh. AB - This paper uses prospective data from the Matlab surveillance system in rural Bangladesh to demonstrate that initially co-resident spouses and sons have a major impact on the subsequent mortality of old people, with significant differences by the sex of the elderly person, and the age of the son. Spouses significantly reduce mortality by similar magnitudes for both elderly men and women. On the other hand, co-resident adult sons reduce mortality for elderly women much more than for elderly men, with younger sons being more beneficial than older sons. Furthermore, both married and unmarried females appear to benefit equally from co-resident adult sons. Finally, this analysis suggests that the impact of spouses and sons on mortality in old age is not substantially mediated through changes in elderly economic status. PMID- 10676062 TI - Provider knowledge about emergency contraception in Ghana. AB - In 1996, the Ministry of Health in Ghana included emergency contraception (EC) in its newly issued National Reproductive Health Service Policy and Standards. A short survey was conducted in the summer of 1997 to evaluate health providers' knowledge of EC. Of the 325 providers interviewed, about one-third (34%) had heard of EC. No provider had sufficient knowledge to prescribe EC correctly. A well-coordinated training programme for providers will have to precede successful introduction of EC in Ghana. Moreover, a dedicated product may be critical for the successful introduction of EC in a country like Ghana, where provider knowledge is low. PMID- 10676063 TI - Contraceptive failure: levels, trends and determinants in Matlab, Bangladesh. AB - This study investigated the levels, trends and determinants of contraceptive use failure in Matlab, Bangladesh, using a set of prospective data on 25,960 women of reproductive age. The data were extracted from the Record Keeping System (RKS) of Matlab for the period 1978-94. If there was any live birth during the use or within 7 months after the discontinuation of use, it was considered as a failure. The life table technique and hazard model were used as analytical tools. The results suggest that use-failure for pills, IUDs (TCu 200) and injectables and other temporary methods increased from 1978 to 1988, but began to decline after 1988. The cumulative probability of first-method failure within 1 year of method acceptance of the cohort of 1990-94 acceptors was 12.9% for pills, 2.0% for IUDs, 0.5% for injectables, 22.0% for condoms and 13.4% for 'other' methods (sampoon, foam, jelly and traditional methods). For pills, condoms and 'other' methods, the likelihood of failure declined with the duration of use; by contrast, the probability of an IUD failure increased over time, peaking at 3 years of use. The injectables maintained a low likelihood of failure regardless of the duration of use. The quality of Community Health Workers' (CHWs) performance was associated with the risk of failure of all temporary methods except condoms; women's background characteristics associated with failure varied by method. The effect of the quality of the CHWs' performance and the background variables on failure did not change much over time. It is felt that contraceptive failure deserves the serious attention of programme managers and policy makers to make the Bangladesh national family planning programme more successful. PMID- 10676064 TI - Fertility and population policy in two counties in China 1980-1991. AB - A survey of women in two highly developed rural counties of China, Sichuan and Jiangsu Provinces, was carried out in late 1991, to gain information about demographic and economic change between 1980 and 1990. Three separate surveys were conducted: the first a questionnaire administered to married women aged 30 39, eliciting information about childbearing and contraception, as well as the social and economic background of the respondents; the second, focus group interviews emphasizing the motivation for childbearing. Official information about the selected villages, townships and counties was also collected. National level data in 1987 show that individual reproductive behaviour in China failed to conform to a universal, effectively implemented, population policy. They imply either a spatial range of policies, or great diversity in the demand for children, or perhaps a combination of both. Such diversity in reproductive behaviour is also found in the study area. The purpose of the analysis was to examine the diversity in reproductive behaviour and contraceptive practice, and to discover whether differentials are influenced by area, or else exist between individuals within areas. If the former, then the explanation may be found in differences in policy formulation and implementation between areas: and if the latter, to demand for children, or else differential application of policy restrictions. The main findings were that: (1) the explanation of the pattern of fertility and contraceptive use is to be found at the individual level (within locations) rather than in policy differences between administrative units; (2) the association between income and number of children is negative, as is that between income and the propensity for uniparous women to remain unsterilized. The theory that privilege may be exercised to gain concessions from birth planning cadres is therefore not supported; (3) ideal family size differentials are largely absent, showing that social (education) and economic (income, occupation) characteristics are not responsible for differences in reproductive motivations, and implying that the nature of the demand for children is very different from that in most rural areas of the Third World; (4) data on ideal family size by sex of the existing offspring indicate only a weak preference for sons. The low demand for children, and the weak son preference, may both be explained by the social acceptability of uxorilocal marriages, and of village endogamy, together with the prohibitive costs of children, and especially of sons. This partly results from the expense of education, but most mothers emphasize marriage costs. It is speculated that the circumstances responsible for the escalating costs of children in the two countries are likely to pertain in growing areas of the country, with the privatization of education and health services, the declining support of collective institutions, and the replacement of this function by kinship networks. These on-going changes imply that any policy of reproductive restriction for the purposes of population control is likely soon to meet with diminishing resistance; and it may later be rendered unnecessary in the eyes of government officials, as fulfilled reproductive intentions lead to a fertility level below replacement level. PMID- 10676065 TI - Crisis intervention strategies when caring for families of children with cancer. AB - A diagnosis of childhood cancer is an unexpected life event that often precipitates a situational crisis for all family members. Required cancer treatments and other ongoing stressors for both child and family will significantly disrupt the family's equilibrium and well-being. An increasingly important role of the pediatric oncology nurse is to facilitate crisis intervention strategies that help families adjust to the psychosocial stresses associated with childhood cancer, yet many nurses have little or no training in crisis theory and/or crisis intervention strategies. This article reviews family crisis theories and outlines crisis intervention strategies that are appropriate for the family of a child with cancer. PMID- 10676066 TI - Choices and control: parental experiences in pediatric terminal home care. AB - During the past decade, palliative care at home has become an alternative option to hospital care for terminally ill children. This study describes the experience of caring for a dying child at home from a parent's perspective. A qualitative research design was used to conduct and analyze data. Nonstandardized, focused interviews were conducted with 10 families. Thematic content analysis assisted in deriving themes from the transcripts of the interviews. "Choice and control" was the major theme that linked all the other concepts, and it appeared to be fundamental to parental coping strategies. Most parents were willing to take responsibility for the nursing care of their child, including administration of intravenous medication. The patient's home was the overwhelming choice of parents for delivery of terminal care, with most parents perceiving it as their child's choice also. PMID- 10676067 TI - Children's perceptions of pain during 3 weeks of bone marrow transplant experience. AB - Most patients undergoing bone marrow transplant (BMT) experience severe pain. Because self-reporting is the most reliable source when assessing pain, it is important that health care providers understand how children perceive their pain and alleviating factors. The purpose of this descriptive, exploratory study was to understand children's perceptions of: (a) their BMT pain, (b) interventions effective in relieving their pain, and (c) caregivers' role in managing their pain. The sample consisted of 20 children (50% male), age 5 to 17 years, undergoing BMT. All study participants received continuous-infusion opioid therapy with additional boluses as needed for pain. Using investigator-developed structured interview guides, children were interviewed four times: on the day of transplant, then at three weekly intervals. Data were analyzed using a content analysis approach. In the first interview, children reported that when they hurt, they most commonly told someone. Several children used nonpharmacologic techniques to relieve pain. Although all had been told to expect some pain during BMT, only one-third of the children described the kinds of pain that they anticipated having during BMT. During subsequent interviews, the majority said that medication worked best to decrease their pain. In the final interview, most children said they still hurt. They reported pain predominantly in their mouth and throat, but mentioned seven other pain sites. Anecdotal comments included that pain management should be improved on the first BMT day and that nurses need to teach children that it is okay to use drugs for pain. Implications for clinical practice, education, and research are discussed. PMID- 10676068 TI - Description of a multihospital process to develop a care path for the child with acute lymphoblastic leukemia. AB - This article describes the National Association of Children's Hospitals and Related Institutions (NACHRI) collaborative group process used to create a multihospital care path for the child with acute lymphoblastic leukemia (ALL), and presents strategies for implementation and future direction. Although most children in the United States with cancer are treated according to National Cancer Institute-sponsored comprehensive protocols, there is a wide variation in the implementation of protocols by physicians and hospitals. The development of this care path was based on evidence from the literature, review of practice patterns, expert opinion, and group participant consensus building. The resulting 4-day care path was organized into six categories of care (e.g., assessment practices, diagnostic tests, teaching, and discharge planning). Discharge criteria are stated at the beginning of the care path to emphasize the planning process immediately on admission. Clinical outcomes, skill and knowledge outcomes for the parent and child, and home assessment considerations are also included. Strategies to create change and gain support of various stakeholders toward implementation of the care path are presented. The strength of the resulting care path is possible in large part because the multihospital group process brought professionals from around the country together to discuss, analyze, and reach consensus on the practices related to the child with ALL. The group process enabled the development of a care path that goes beyond a traditional care path developed by a single institution. PMID- 10676069 TI - Home care requirements for children and adolescents with cancer. National Association of Children's Hospitals and Related Institutions (NACHRI) Patient Care Oncology FOCUS Group. PMID- 10676071 TI - A nurse's roadmap. PMID- 10676070 TI - Radiation somnolence syndrome. AB - Although initially described 7 decades ago, somnolence syndrome remains a poorly understood subacute effect of cranial irradiation. Despite the relatively transient and benign nature of somnolence syndrome, its symptoms can be distressing for children and caregivers. Anticipatory guidance related to radiation-induced somnolence remains a critical nursing intervention. This article reviews what is known about somnolence syndrome, including its causes, symptoms, and management. PMID- 10676072 TI - Testability of a color vision screening test in a population with mental retardation. AB - PURPOSE: The purpose of this study was to determine the testability of the "Co or Vision Testing Made Easy" color vision test, marketed as a screening test for young children, in a population of individuals with mental retardation. The test uses simple geometric figures that are easily identified. Previously, the test has demonstrated validity as a measure of color deficiency. METHODS: The test was presented to Special Olympic athletes, who are individuals with mental retardation or significant developmental delay, at four sites: the 1997 World Winter Games in Toronto, Canada; the Texas Summer Games in Houston, Texas; the Massachusetts Summer Games in Boston, Massachusetts; and Regional European Swim Competition in Seville, Spain. The criteria for passing was 8 correct responses on the first trial or 9 of 9 on the second attempt. RESULTS: Testability in Toronto, Canada; Houston, Texas; and Seville, Spain was high--95.5%, 98.7%, and 95.7%, respectively. Testability, however, dropped to 78.8% during the Boston, Massachusetts screening. There was no apparent difference in the testing environment that would account for the difference. The overall rate of testability was 93.2% for the 1078 athletes screened. The frequency of males identified as color deficient was similar to that expected in the general population; only two females (in Spain) failed the color vision screening. CONCLUSIONS: The "Color Vision Testing Made Easy" color vision test was successfully completed by a very high percentage of Special Olympics athletes. These results suggest that this test is useful in screening this population for color deficiencies, and that the prevalence of color vision deficiencies is approximately the same in individuals with mental retardation as in the general population. PMID- 10676073 TI - Nearpoint phoria changes associated with the cessation of childhood myopia progression. AB - BACKGROUND: A convergent (eso) shift in near phoria associated with the onset of myopia has been reported. METHODS: Data from two Midwestern United States optometry practices were used to assess whether the near phoria shifted back in the divergent (exo) direction after the cessation of childhood myopia progression. Data were collected for myopic children who had three or more examinations before the age of 15 years and at least one examination after the age of 17 years. RESULTS: Refractive error data were used to calculate an index of the age of cessation of childhood myopia progression. The phoria at the first examination after the cessation age was designated as the baseline and was normalized to zero. For all previous and subsequent examinations, the changes in phoria with respect to the baseline phoria were calculated. The phoria at the examination just after the cessation age was significantly more divergent than those at the first, third, and fourth examinations prior to the cessation age (1.1, 1.4, and 1.7 prism diopters, respectively). The third visit after the cessation age was 1.8 prism diopters more divergent than the first visit after the cessation age. Thus, these data showed an exo shift in near phoria after the cessation of childhood myopia progression. PMID- 10676074 TI - Keratoconus with good unaided visual acuities: two case reports. AB - BACKGROUND: Ophthalmic evaluation of patients with keratoconus (KC) often reveals highly myopic and irregular astigmatic refractive corrections. Irregular corneal astigmatism and central corneal scarring in patients with KC often result in a loss of best-corrected spectacle acuity. Rigid gaspermeable contact lenses generally optimize visual acuities for patients with KC. CASE REPORTS: Two cases are discussed of patients who manifested clinically diagnosed KC but unusually good unaided Snellen visual acuities (20/25+ or better) in both eyes. CONCLUSION: Good unaided visual acuities are not necessarily inconsistent with the diagnosis of KC. PMID- 10676075 TI - Circumscribed posterior keratoconus: case report. AB - BACKGROUND: Posterior keratoconus has only a few scattered case reports in the literature. Posterior keratoconus is characterized by a posterior stroma thinning and a depression of the posterior corneal surface. The effect on acuity is variable and may be related to other ocular and systemic conditions. CASE REPORT: An African-American woman came to us with posterior concavity (corneal thinning), with stromal scarring in both eyes and an epithelial iron ring present in the left eye. The endothelia layer appeared intact in both eyes. Corneal topography of the right eye demonstrated a central flattened zone with peripheral steepening, while the left eye an inferior nasal steepened zone was present. The patient was also diagnosed with myopic degeneration (O.D. > O.S.) as well as cataracts. CONCLUSIONS: Posterior keratoconus generally has a minimal effect on visual performance and requires no specific treatment. In cases in which visual defect is severe and is attributable to the posterior keratoconus--and not other ocular conditions, such as cataracts--penetrating keratoplasty should be considered. PMID- 10676076 TI - Late traumatic intraocular lens extrusion after penetrating keratoplasty. AB - BACKGROUND: Penetrating keratoplasty places a patient at risk for wound rupture from blunt trauma because the graft-host interface remains weakened for years after the surgery. Violent environments, contact sports, and strenuous activity put patients with compromised corneal structural integrity at high risk of traumatic injury. CASE REPORT: This case report presents a 42-year-old penetrating keratoplasty patient with a history of homelessness, polysubstance abuse, and domestic violence. This patient experienced a ruptured globe at the graft-host junction secondary to a direct blow by a fist, which extruded the intraocular lens from the eye. After emergency wound closure, the graft continued to degrade until bullous keratopathy developed. With little visual recovery potential for this graft, a Gunderson conjunctival flap procedure was implemented to decrease chronic ocular pain. CONCLUSIONS: After penetrating keratoplasty, patients should be periodically reminded of the susceptibility of the graft wound to injury from high-risk activity and violence. Constant use of protective eyewear should be recommended to corneal transplant recipients. PMID- 10676077 TI - Hiring a new optometrist? Be prepared! AB - Bringing a new optometrist into a practice, even if only on an employer-employee basis, is a complicated process that should not be taken lightly by hirer or hiree. This article explores the many aspects of the process and the relationship that must be understood and worked out in advance. PMID- 10676078 TI - Staying in shape: the different roads to fitness for busy optometrists. AB - Do you keep telling yourself it's time to get in shape? Are you adding workouts to your list of New Year's resolutions? If you think you can't balance fitness with a busy schedule, think again. Here are the stories of some optometrists who have done just that (along with a couple of hard-core ODs who've taken fitness to a higher level). PMID- 10676079 TI - Should there be policies to restrict visitors during labor and birth? PMID- 10676080 TI - The Cool Kids Coalition. AB - The Cool Kids Coalition was initiated as a community response to more than 214 hospitalizations of children under the age of five for burns over a 6-year period in one township in Long Island, NY. The coalition was started by public health nurses in partnership with the local chapter of the National Safe Kids Campaign. Goals included: 1. parent education regarding scald burn prevention; 2. development of innovative interventions for those at risk; and 3, development of innovative community approaches to scald prevention. Coalition members had diverse backgrounds and the coalition integrated non-traditional partners in injury control. The coalition doubled in size due to overwhelming community interest, growing within a few months from an initial group of 15 to a well represented group of 30. Innovative programs were implemented that reached more than 3,000 parents, both in the community and home. Teaching was conducted with parents in the target population in Head Start centers, homeless shelters, the home, libraries, child care centers, a shelter for teen parents, etc. Member agencies incorporated the booklet and materials into their individual programs. The development of the Cool Kids Coalition illustrates the power of nursing in community health. PMID- 10676081 TI - A nurse practitioner model of practice in the neonatal intensive care unit. AB - PURPOSE: The purpose of this study was to describe the practice of the nurse practitioner (NP) in the neonatal intensive care unit (NICU) in an attempt to define an advanced practice nursing model that is unique to NP practice in the NICU. DESIGN: This qualitative research used an ethnographic case study design to answer the research question: 'What is the practice model of nurse practitioners working in the NICU?' METHODS: Seven nurse practitioners working in five level II/III NICUs in Massachusetts and Rhode Island were interviewed and observed in practice. Audiotaped interviews using open-ended questions and field notes from participant observations were analyzed for patterns of behavior. RESULTS: These nurse practitioners practiced within a model of advanced practice nursing that emphasized health, holism, and caring within the highly technological and medical NICU environment. CLINICAL IMPLICATIONS: A model of NP practice in the NICU is emerging and needs further development and testing. Nurse educators and administrators must find ways to support the nursing model in the advanced practice nursing role in the NICU. Nurse practitioners need to be more active in promoting a clearer understanding of their practice and contributions to the NICU care delivery team. PMID- 10676082 TI - Investigating the relationship between satisfaction with social support and functional status after childbirth. AB - PURPOSE: To explore the relationship between satisfaction with support and functional status after childbirth. DESIGN: Prospective longitudinal survey design. METHODS: Two hundred new mothers who had experienced a healthy pregnancy, normal delivery and puerperium, and delivered a healthy infant between 37 and 42 weeks gestation were approached while attending primary healthcare maternal-child health centers and immunization clinics in New South Wales, Australia. Measurement tools included the Inventory of Functional Status After Childbirth and the Support Behavior Inventory, and were administered at 6 weeks, 3 months and 6 months. RESULTS: Significant increases in total mean functional scores, scores for household responsibilities, social activities, and self-care were noted. Although no significant correlation was found between satisfaction with social support and functional status after childbirth, satisfaction with support from one's partner decreased significantly during the 6-month survey period, as did satisfaction with support from others. CLINICAL IMPLICATIONS: It could be that providers need to assess the social support needs of their clients. A postnatal support plan could be used by mothers to negotiate the long-term involvement of others in household tasks and selected aspects of infant care. PMID- 10676083 TI - African American mothers use stories for family sexuality education. AB - PURPOSE: To examine the sexual knowledge and cultural values transmitted by stories from African American mothers to their adolescent daughters. METHOD: Narrative analysis. Stories from 11 mothers were recorded and transcribed by entering interview narratives into Qualpro, a computerized data management program, and examined using a narrative analysis method. RESULTS: Analysis revealed that mothers used story telling as a strategy for family sexuality education. The mothers used stories from their own experiences to accomplish socialization/enculturation and to discourage their daughters from making the same mistakes that they reportedly made (such as becoming pregnant during the teenage years). Findings supported the fact that stories served as cultural artifacts that describe the cultural pathways of a group of African Americans mothers and daughters. IMPLICATIONS: It is important for nurses to have an awareness of the importance of ethnocentric models for community health nursing practice and culturally sensitive assessment tools for adolescent sexuality education programs. PMID- 10676084 TI - Maternal stress during hospitalization of the adopted child. AB - PURPOSE: To identify and describe the experiences of mothers whose adopted children are hospitalized and to compare those experiences to those of mothers whose biological children are hospitalized. DESIGN: Comparative descriptive design. METHODS: Mothers of hospitalized children (n = 33 adopted; n = 19 biological) completed a slightly revised version of the Parental Stressor Scale: Pediatric Intensive Scale Unit (PSS:PICU). Adoptive mothers also completed a questionnaire related to their perceptions of the impact of their child's adoption on the hospitalization experience. RESULTS: Adoptive mothers perceived statistically significantly higher levels of stress related to their child's behavioral/emotional response to hospitalization. Children who had been with the family for less time perceived the hospitalization as more stressful. When both groups of mothers were considered, mothers of younger children perceived a higher level of stress. A majority of adoptive mothers felt the adoption had an impact on the hospitalization experience, especially related to their limited information about their child's medical history, staff lack of knowledge about legal issues, and concern about attachment to the child. CLINICAL IMPLICATIONS: Nurses need to be aware of adoptive mothers' concerns related to attachment issues, limited family medical history, and legal rights in order to provide sensitive and effective care. Inservice education programs could be designed to help teach all staff about these important issues. PMID- 10676085 TI - Perceived barriers to prenatal care services. AB - PURPOSE: The purpose of this study was to determine barriers to prenatal care services and to determine if barriers differed by demographic characteristics in a low-income population. DESIGN: Descriptive correlational study with 110 women who sought prenatal care after the 20th week of gestation. RESULTS: Two items were major barriers to seeking prenatal care: long waiting times at the time of appointments and the cost of getting care. Significant relationships were found based on the age and race of the women. CLINICAL IMPLICATIONS: Some identifiable variables prevented these women from seeking early prenatal care; however, the barriers identified are amenable to change. Strategies to reduce barriers could include providing more culturally competent care, more timely appointments, better use of the woman's time when appointments are kept, educating women in the community about the availability of low-cost care, and assistance at prenatal care sites for facilitating completion of insurance and financial applications. Barriers to prenatal care varied by demographic group; therefore, identifying the characteristics of the group being served seems important in efforts to decrease barriers to care. PMID- 10676086 TI - So you want to be an expert witness? Things you need to know. PMID- 10676088 TI - Evidence-based practice. PMID- 10676087 TI - Family liaison programs. PMID- 10676089 TI - Workshop on medical education. PMID- 10676090 TI - Medical education at the present time. PMID- 10676091 TI - Educational innovations in the medical faculty. PMID- 10676092 TI - Telemedicine and processing of scientific information in programs of continuing education. AB - Starting from some reflections on the goals of medical education, central questions are analyzed in relation to the transfer of scientific knowledge and the generation of more advanced cognitive approaches in professionals who follow distance courses. The core of this article refers to the information management processes, according to a cognitive perspective, in terms of the development of the medical thought. With that intention, particular educational instruments are indicated in order to achieve the internalization of the "modes of knowledge" acquired through significant learning. The basic criterion of the educational program is that physicians can best operate personally in the teaching processes relatively to their particular scientific domain, because they know, better than anyone else, what the proper methods of knowledge are: epistemology is the basis of any special teaching, the article makes specific reference to the production of texts of information technology. However, the general approach is applicable to any other means that may be used in telemedicine. PMID- 10676093 TI - Medical education: what can be learned from Europe. PMID- 10676094 TI - Medical education in Spain. PMID- 10676095 TI - Scientific research policy: elements for comparing Europe and Italy. PMID- 10676096 TI - University and enterprise: the future of knowledge and research, the future of work. PMID- 10676097 TI - The university and the needs of the society: what professional models? AB - An overview of the complex Italian situation and the consequences of the European integration, which puts an emphasis on the role of research and education involving the universities and the specific faculties, is given. Attention is then focused on the characteristics of the evolution of the medical activity and health services as for prevention, health education, more extensive knowledge and the need for continuing education. Different problems are tackled and pertinent suggestions are offered. The cultural and professional perspectives of the doctor should be considered within a new psychosocial approach to the illness, super- and hyper-specialization, collaboration and skills in non traditional fields. Medical education should be based on tutorial teaching and student-centered rather than on the traditional teacher centered-academic teaching. For better health care medical education and training should be updated with respect to the doctor-patient relationship as well as to the technological advances and team work in medicine. The ethical aspects of the medical profession should be evidenced to be able to tackle the involved problems. The main features of the doctor of the future are the need and the difficulty of updating and life-long learning. PMID- 10676098 TI - Continuing education towards self-education. PMID- 10676099 TI - Teaching the teachers. AB - The author analyzes how the problem is dealt with in Italy in comparison with other European countries. He also recalls the early moves taken in Italy for teachers' training, very often started outside the university. The possible ways of intervention are discussed for the improvement on the present situation which, as a whole, is considered unsatisfactory, in spite of commendable individual initiatives in a few Italian medical faculties. PMID- 10676101 TI - The "Universita Cattolica del S. Cuore" and its educational offer. PMID- 10676100 TI - Guidelines for medical education at the Medical Faculty of the Catholic University of Lille, France. PMID- 10676102 TI - [Guidelines: research in the web, critical assessment, clinical application]. PMID- 10676103 TI - [New trends in pharmacotherapy of arterial hypertension]. PMID- 10676104 TI - [Treatment of acute myocardial infarction: present and future]. PMID- 10676105 TI - [Educational status, social class indicators and diseases]. PMID- 10676106 TI - [Mechanism of action of quinapril in the treatment of primary arterial hypertension]. AB - The effects of a long-term therapy with quinapril on plasma renin activity, plasma aldosterone, atrial natriuretic peptide and left ventricular mass were analysed in patients with mild to moderate systemic hypertension. Fifteen patients (4 women) were treated for one year with quinapril 10 or 20 mg once daily, reducing hereby the systolic and diastolic blood pressure from 167.5 +/- 11.3 to 141 +/- 6.7 mmHg p < 0.001 and from 105.3 +/- 5 to 90 +/- 7 mmHg respectively, within the first two weeks. Blood pressure remained stable during the following 52 weeks. After 6 and 52 weeks of therapy, as expected, we observed an increase of plasma renin activity, plasma aldosterone decrease from 262.6 +/- 88.1 to 178.8 +/- 79.9 p = 0.01 and to 170.3 +/- 64.3 ng/ml p = 0.006 respectively. Atrial natriuretic peptide levels were not significantly altered. After 52 weeks of treatment left ventricular mass index decreased from 107.9 +/- 16.2 to 90.1 +/- 13.4 g/m2 p = 0.0001. It is concluded that treatment with quinapril for 1 year in addition to controlling blood pressure also reduced left ventricular mass probably by a favourable effect on renin-angiotensin-aldosterone system. PMID- 10676107 TI - [Amiodarone pulmonary toxicity]. AB - The Authors discuss a case of amiodarone pulmonary toxicity, with simultaneous alveolar and interstitial infiltrates, in a female patient 75 years old, who took the drug for 6 months at a low dosage (200 mg/daily for 5 days a week). It is outlined that the diagnosis can be achieved only by exclusion of other aetiologies, since clinical and diagnostic features are not pathognomonic for such disease. The withdrawal of the drug and the administration of the steroid therapy determined a fast improvement of the clinical and radiological appearance. PMID- 10676108 TI - [Mycosis fungoides: a clinical case]. PMID- 10676109 TI - [Guidelines in internal medicine]. PMID- 10676110 TI - [Clinical diagnosis of infective endocarditis]. PMID- 10676111 TI - [Treatment of infective endocarditis]. PMID- 10676112 TI - [Endocarditis in valve prosthesis]. PMID- 10676113 TI - [Infective endocarditis in the elderly]. PMID- 10676114 TI - [Nephropathy: current aspects and therapeutic prospects]. PMID- 10676115 TI - [Cardiorenal physiopathology and heart failure: current aspects and therapeutic prospects]. AB - The heart and the kidney exert a reciprocal control of their function in order to maintain a steady state of haemodynamics, both in physiological and pathological conditions. The functional relationship between the two organs becomes particularly evident during heart failure. The knowledge of such relationship may play an important role in the management of heart failure. We also report here our experience in the treatment of congestive heart failure with depletive techniques vicarious of kidney function. PMID- 10676116 TI - [The third scientific revolution]. PMID- 10676117 TI - [Health as a fundamental human right: the development of the World Health Organization]. PMID- 10676118 TI - [Evolution of health systems]. PMID- 10676119 TI - [Peace, vaccine and potatoes (the decline of high mortality]. PMID- 10676120 TI - [Vaccination policies and the eradication of smallpox]. PMID- 10676121 TI - [Medicine and molecular biology: a happy marriage?]. PMID- 10676122 TI - [Medically-assisted reproduction]. PMID- 10676123 TI - [Neonatology: several history lessons]. PMID- 10676124 TI - [The arrival of organ transplantation]. PMID- 10676125 TI - [The major phases of medical imaging]. AB - In this article, we give an insight into the historic developments of radiology, evoking some of the milestones which from the end of the 19th century to the beginning of the third millennium have marked the way of one of the most rapidly growing fields in medicine. PMID- 10676126 TI - [Blood and transfusion]. PMID- 10676127 TI - [Emergence of infectious diseases]. PMID- 10676128 TI - [Turning points in psychology and psychiatry in the 20th century]. PMID- 10676129 TI - [Rational union between medical sociology and academics]. PMID- 10676131 TI - [The paradox of heads and tails]. PMID- 10676130 TI - [Medicine, bioethics, public health]. PMID- 10676132 TI - [The correspondence of Gottfried Brugger (1857-1891) or the tragic fate of a young veterinarian]. PMID- 10676133 TI - [A student circle of friends: the portrait collection of the veterinarian Gottfried Pfister (1836-1903)]. PMID- 10676134 TI - [Formation of veterinarians from Vaud in the 18th century]. PMID- 10676135 TI - [Guide dogs for the blind: aspects of a special human-animal relationship in history and the present time]. PMID- 10676136 TI - [Memory advantage of performed actions: comments on multimodal memory theory]. AB - Based on the integration of the memory advantage for subject-performed actions into the multimodal theory of episodic memory by J. Engelkamp (1997), three issues referring to the so-called enactment effect are discussed and addressed by statistical r-analyses. Firstly, the empirical basis of the functional distinction between motor and non-motor memory resources by means of dual-task experiments is questioned. Secondly, a multinomial modeling analysis is presented which aims at the contributions of automatic and controlled memory processes to the enactment effect in the process-dissociation paradigm. Finally, the effect of enactment on memory for serial order information is discussed with respect to recent accounts of serial memory. PMID- 10676137 TI - [Selective interference, process dissociation and serial information in recall of performed actions. A comment on Thorsten Meiser's remarks on the multimodal memory theory]. AB - In this commentary, I deal with three questions. (1) Do the findings of interference experiments justify the distinction between motor and visual imaginal processes? (2) Can automatic and controlled process components be identified by the process-dissociation technique? (3) Does encoding of order information depend on encoding instructions? PMID- 10676138 TI - [Effects of knowledge acquisition on verbal linearization about spatial orientation]. AB - This paper deals with determinants of linearization in spatial communication. The well-known linearization principles, such as the principle of natural order, primarily emphasize features of the given information. We assume that verbal linearization is in addition influenced by the speakers' knowledge acquisition. Six experiments with a total of 272 participants are reported here. A particular spatial constellation was presented to the participants, who were asked to talk about it afterwards. The linearizations then produced were defined as dependent variables. The results show that the linearizations were influenced by the speaker's experience with the configuration (according-to-experience effect) and also demonstrate the importance of the first encounter (anchor effect). Further characteristics of the effects mentioned above are described and discussed regarding memory research and psycho-linguistics. PMID- 10676139 TI - [Parametric coupling in sequences of bimanual reversal movements with identical and different amplitudes]. AB - In sequences of bimanual movements, the voluntary modulation of the amplitude of the one hand (inducing hand) induces an involuntary modulation of the amplitude of the other hand (dependent hand), the strength of which increases with increasing tempo. By means of a task in which subjects perform sequences of two short and two long reversal movements in alternation with the inducing hand, but constant short or long reversal movements with the dependent hand, we addressed two questions. The first question was concerned with differences in the effects of tempo on the involuntary amplitude modulation of short and long movements; the second question was whether the involuntary amplitude change fades away or is propagated when bimanual movements with certain target amplitudes are repeated. At low tempo the contralateral effect of voluntary amplitude changes on short amplitude movements was stronger than the effect on long-amplitude movements, but at high tempo this difference was reversed. This result is not consistent with the assumption that contralateral amplitude modulation results from an overflow of efferent commands, which increases with the force of the movement; however, it is consistent with other findings on a transient coupling during amplitude specification (parametric coupling). The involuntary amplitude change was essentially propagated to the next movement in the sequence and did not fade away. This finding suggests that the assimilation of amplitudes that can be observed in bimanual sequences of movements with different, but for each hand constant, amplitudes could result from an effect of transient parametric coupling during the initial specification of amplitudes and need not necessarily be attributed to cross-talk at the level of motor commands or efferent innervation. PMID- 10676140 TI - [Recognize the victim in need: the effect of situational cues and priming on identification of need for help]. AB - An experimental study investigated the joint influence of priming and situational cues on the perceived neediness of a target. A scrambled sentence test was used to prime two groups with the concepts of neediness and safety. In a third group a neutral concept was activated. A free description of the target and a subsequent adjective rating were applied as dependent measures. Although results indicated that both factors affected the attribution of neediness on a free description, the effects of priming were only apparent on this measurement and disappeared on the second task. PMID- 10676141 TI - [Posner's theory of attention: right hemisphere processing advantage in extended attention]. AB - Posner proposed a theory of attention based on target detection, visual orienting, and alertness. The latter is supposed to use brain structures mainly located in the right hemisphere. Whitehead found a right hemisphere processing superiority during sustained attention. An additional auditory stimulus is thought to produce a change of alertness and should interfere with this asymmetry, which Whitehead was able to show, too. It remains unclear how the left hemisphere is activated by right hemisphere pathways. Therefore we tried to replicate Whitehead's findings. In our first experiment the expected interaction between visual field, foreperiod duration, and tone was obtained. Probably the tone used in our experiment was not intense enough to produce a sufficient change in alertness. We used a more intense tone in a second experiment. This time a three-way interaction was present but could not be interpreted in terms of Whitehead's assumptions. Instead, the additional alerting stimulus seems to influence the state of alertness in a much more general and long-lasting way. PMID- 10676142 TI - [Effects of dimensional and social comparisons on ability assessment and satisfaction with performance]. AB - The study deals with the question whether internal dimensional comparisons (comparisons of one's own achievement in a task with one's own achievement in another task) contribute to the development of task-specific self-evaluations. In an experimental study, N = 135 student subjects who had worked on two different types of tasks received manipulated achievement feedback with reference to dimensional and social comparisons. Results indicated the relevance of dimensional comparison information. Subjects scored their own ability and contentment with their result in the first type of task more positively (negatively) when they had received a better (worse) result than in the second task. PMID- 10676143 TI - Epidemiology of feline leukaemia and feline immunodeficiency virus infections in the Czech Republic. AB - Commercial serological sets were used for the examination of 727 cats kept in larger towns of the Czech Republic. FeLV antigen and antibodies to FIV were demonstrated in 96 (13.2%) and 42 (5.8%) of the animals, respectively. Seven (0.96%) animals were positive for both FeLV and FIV. Most of the FeLV and/or FIV positive patients were intact rambling males aged 1-4 years. Chronic gastrointestinal and respiratory diseases were found in 54.2% and 43.8% of the FeLV-positive patients, respectively. Chronic urinary tract diseases and generalized lymphadenopathy were found in 47.6% and 45.2% of the FIV-positive patients, respectively. The results of this first survey in the Czech Republic have shown prevalence values and clinical patterns similar to those reported formerly from other European countries. PMID- 10676144 TI - Prevalence and seasonal pattern of caprine trichostrongyles in a dry area of central Spain. AB - A total of 322 gastrointestinal tracts were examined from traditionally reared goats originating from a dry area of central Spain. A large spectrum of gastrointestinal nematodes was observed, Teladorsagia circumcincta and T. trifurcata being the most prevalent species, followed by Trichostrongylus vitrinus and Nematodirus filicollis. Trichostrongylus capricola recorded a high level of infection despite low prevalence. A nematode infection rate of 93% and an average burden of over 3000 worms indicates the highly prevalent infection in Spanish goats. The seasonal pattern shows a real risk of infection all year round, with two peaks in the summer and autumn. Females were more frequently infected than males. PMID- 10676145 TI - Aggregation of sow lactobacilli with diarrhoeagenic Escherichia coli. AB - A total of 20 strains of lactobacilli were isolated from the oesophagus and vagina of 20 sows at the time of partus. Aggregation activity was seen between six homofermentative autoaggregative lactobacilli and three strains of pathogenic Escherichia coli with F4, F5 and F6 fimbriae. The highest aggregation activity was observed between vaginal Lactobacillus acidophilus PV 32 or oesophageal OE 2/1 and E. coli with F4 (K88). The presence of aggregation-promoting factor (APF) was confirmed by polymerase chain reaction (PCR) with primers of a specific fragment the apf gene derived from human L. gasseri 4B2 in one oesophageal L. acidophilus strain OE 2/1. We propose that autoaggregative lactobacilli that aggregate with diarrhoeagenic E. coli can express a class of APF proteins that exhibit the function of an aggregation mediator. PMID- 10676147 TI - Pathological and microbiological studies on pneumonic lungs from Danish calves. AB - During 1 year, the association between microbiological and pathological findings in 72 lungs from calves submitted to the Danish Veterinary Laboratory for diagnostic purposes was studied. All cases were evaluated pathologically and bacteriologically, whereas only 68 cases were examined for the presence of bovine respiratory syncytial virus (BRSV), parainfluenza-3 virus (PI-3 virus) and bovine coronavirus, 62 cases for bovine viral diarrhoea virus (BVD), 45 cases for bovine adenovirus and 51 cases for mycoplasmas. Based on histopathological examination, the cases were diagnosed as fibrinous and/or necrotizing bronchopneumonia, suppurative bronchopneumonia, embolic pneumonia and others. The diagnoses were based on the dominating and most severe lesions in each lung. Haemophilus somnus, Pasteurella multocida, Actinomyces pyogenes, P. haemolytica and BRSV were the most commonly found bacterial and viral lung pathogens, respectively. Pasteurella spp. and H. somnus were often associated with the more severe fibrinonecrotizing type of bronchopneumonia, whereas BRSV was primarily detected in cases of suppurative bronchopneumonia. Mycoplasma bovis was isolated from one case only, whereas M. dispar, M. bovirhinis and Ureaplasma diversum were present, often concomitantly, in the majority of cases. Aspergillus fumigatus was isolated from one case. PMID- 10676146 TI - Pigeon paramyxovirus-1 (P-group) as the cause of severe outbreaks in fancy Columba livia in Saudi Arabia. AB - An avian paramyxovirus-1, which was previously isolated from ailing fancy Columba livia in Saudi Arabia, was characterized in this study using monoclonal antibodies (Mabs). The results indicate that although the virus belongs to the pigeon paramyxovirus-1 (PPMV-1) group 'P' still it shows some variation in its binding pattern to the Mabs. PMID- 10676148 TI - Serum bile acids in captive bustards. AB - Blood samples were collected from clinically normal male and female houbara (Chlamydotis undulata macqueenii), kori (Ardeotis kori), buff-crested (Eupodotis ruficrista gindiana) and white-bellied bustards (E. senegalensis) to determine serum bile acid concentrations. Bile acid concentrations were determined by analysis with an Ultrospec 3000 ultraviolet/visible spectrophotometer, using an enzymatic bile acid test. The results provided values of serum bile acid concentrations for the four species, with means +/- standard errors of 35.8 +/- 2.8 mumol; 51.1 +/- 5.0 mumol; 18.4 +/- 2.1 mumol and 20.8 +/- 5.4 mumol for the houbara, kori, buff-crested and white-bellied bustard, respectively. Although no gender or age differences were detected within species, the results demonstrated significant differences in concentrations in clinically normal individuals between the different species. PMID- 10676149 TI - Coagulase-negative staphylococci and mammary gland infections in cows. AB - Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria from bovine mammary gland milk samples. The objective of this study was to determine the type of inflammation evoked by CNS in the mammary gland of cows during their first lactation. Twenty-four Israeli-Holstein heifers in their first lactation were tested for bacteriological status, somatic cell count (SCC) and differential leucocyte count in milk 60-120 days postparturition and every 50-60 days after until drying off. Following the first testing, the 96 quarters of the 24 heifers were classified as follows: 69.8% as no bacterial growth (NBG), 27.1% infected with CNS and 3.1% infected with Staphylococcus aureus. During lactation, 84.5% quarters had no change in their classification, 6.2% were newly infected with other pathogens, 3.1% were classified as self-cured and in 6.2% sporadic bacteria were isolated. Among the CNS, S. intermedius, S. chromogenes and S. haemolyticus were the most frequently isolated. Milk from CNS-infected quarters had significantly higher SCC than milk from NBG quarters. An analysis of the leucocyte pattern in milk from CNS vs. NBG quarters revealed a significant increase in polymorphonuclears and a significant decrease in the percentage of total lymphocytes and lymphocytes bearing CD4+ or CD8+. The high percentage of CNS-infected quarters that remained unchanged in their bacterial status during the first lactation, indicates that those CNS have the ability to elude the immune system and persist in the mammary gland for a long time. The persisting infection, resulting to some extent from an increase of SCC by some CNS strains, suggests that in the near future control steps will have to be taken into consideration, in order to enhance the improvement of milk quality. PMID- 10676150 TI - Efficacy of an intranasal immunization with gEgC and gEgI double-deletion mutants of Aujeszky's disease virus in maternally immune pigs and the effects of a successive intramuscular booster with commercial vaccines. AB - In this study, an intranasal immunization strategy was set up in maternally immune pigs in order to protect them not only clinically but also virologically. Two genetically engineered Aujeszky's disease virus (ADV) strains, Kaplan gE-gI- and Kaplan gE-gC-, were used for intranasal immunization. Both strains were safe for 4-week-old pigs. A single intranasal inoculation of 10(6.0) TCID50 of Kaplan gE-gI- and Kaplan gE-gC- at 4 weeks of age in the presence of moderate titres of maternally derived antibodies (SN titres: 12-16) reduced the amount of weight loss, fever and virus excretion upon challenge 6 weeks later. In a second experiment, the effect of an additional intramuscular booster with three different commercial vaccines (containing attenuated Bartha or NIA3-783 or inactivated Phylaxia; all suspended in an oil-in-water emulsion) at 10 weeks of age was evaluated. One month after the last intramuscular booster, between five and seven pigs from each group were selected for challenge. All intranasally/intramuscularly immunized pigs showed a significantly better clinical and virological protection after challenge than the single intranasally immunized pigs. In the double immunized group, the protection was better when Kaplan gE-gC- was used for the intranasal priming (only two of 14 pigs excreted virus with a duration of 4 days) than when Kaplan gE-gI- was used (13 of 18 pigs excreted virus with a duration ranging from 1 to 4 days). The virological protection was not influenced by the type of vaccine used for booster vaccination. Because the intranasal/intramuscular immunization approach is very compatible with current pig movements on farms and pigs with moderate levels of maternally derived antibodies can effectively be immunized, it can be considered as a good alternative to intramuscular/intramuscular vaccinations especially in regions with a high ADV prevalence. PMID- 10676151 TI - Isolation and further characterization of phase variants of Streptococcus equi subsp. zooepidemicus. AB - In the present study the soft agar technique was used to isolate phase variants of S. equi subsp. zooepidemicus-cultures isolated from infections of horses. The phase variants were characterized by a compact or diffuse colony morphology in this media. The variants could be cultivated separately and further characterized genotypically by RAPD analysis and by macrorestriction analysis of their chromosomal DNA by pulsed-field gel electrophoresis, indicating the identity of both strains of each pair. The diffuse colony variants grew uniformly turbid after cultivation in fluid media, did not haemagglutinate rabbit erythrocytes, and displayed a reduced surface hydrophobicity in hexadecane and phenyl-sepharose adherence tests. The compact colony variants generally grew as sediment with clear supernatant in fluid media, haemagglutinated rabbit erythrocytes and showed an enhanced surface hydrophobicity in both hydrophobicity tests. The presented soft agar technique allowed a demonstration of phase variation of S. equi subsp. zooepidemicus and a subsequent isolation of the variants. This might be an important prerequisite to understanding the pathogenic importance of phase variation among isolates of this bacterial species. PMID- 10676152 TI - Antibody against Testudo herpesvirus is not common in Chinese soft-shelled turtles. AB - Seventy-six serum samples of Chinese soft-shelled turtles (Trionyx sinensis) were collected at Jiangsu Province, China. The neutralization test (NT) was performed with the sera, Testudo herpesvirus (THV) and turtle heart cells (THC). Neutralizing antibodies were detected in five of 76 samples and the titres were 1:10-1:20. Having optimized the conditions, the dot-enzyme-linked immunosorbent assay (Dot-ELISA) was developed and eight serum samples exhibited positive results. Five samples were positive by both NT and Dot-ELISA. The percentage of positive samples was only 6.6% (NT) and 11% (ELISA). It is suggested that THV infection is not a serious problem for the Chinese soft-shelled turtle culture in this region. PMID- 10676153 TI - Bovine herpesvirus 1 (BHV1) seroprevalence in the breeding cattle population of the Veneto region: prospects for the implementation of a control programme. AB - The results of a serological survey for bovine herpes virus (BHV1) antibodies in the breeding cattle population of the Veneto region are presented. The data do not support the hypothesis of an high prevalence of BHV1; on farms where vaccination was not carried out most animals were seronegative, and seropositive animals were generally older. Therefore, when drawing up the guidelines for a control programme, systematic immunization (with glycoprotein E-deleted vaccines) should be restricted only to farms with a high prevalence of BHV1 antibodies and/or with a high risk of BHV1 occurrence; in most unvaccinated farms a 'test and removal' policy appears to be more appropriate in order to rapidly eradicate BHV1 from the entire stock. PMID- 10676154 TI - Mastitis caused by Mycoplasma mycoides subspecies mycoides (large colony type) in goat flocks in Spain. AB - We describe three different outbreaks of mastitis caused by M. mycoides subspecies mycoides LC type (Mmm LC) in three goat flocks from the Extremadura Region of south-west Spain. Thirty-two fast-growing isolates were obtained on Hayflick's and Friis's media with inhibitors from different specimens. All were identified as Mmm LC in spite of their cultural, biochemical and serological features. PMID- 10676177 TI - A strategy for visioning the future for medical education. PMID- 10676178 TI - Community primary care preceptors: does practice site or specialty make a difference? PMID- 10676179 TI - Health professionals' experiences with continuing and distance education. PMID- 10676180 TI - A partnership model for a health professions student pipeline. PMID- 10676181 TI - Changing the culture of "unmatch day". PMID- 10676182 TI - A pre-baccalaureate course on becoming a health professional. PMID- 10676183 TI - Using medical students in a translation service for the hospital. PMID- 10676184 TI - Application of a web-based instruction for a clinical course. PMID- 10676185 TI - A multimedia CD-ROM tool to improve residents' cardiac auscultation skills. PMID- 10676186 TI - The collaborative pediatrics self-study site for on-line self-assessment for medical students. PMID- 10676187 TI - Teaching medical information retrieval skills to internal medicine residents at the terminal-side. PMID- 10676188 TI - Computer-administered formative quizzes in a basic science course. PMID- 10676189 TI - A collaborative virtual medicine library. PMID- 10676190 TI - Using laptop computers for teaching and evaluation in an extended community preceptorship. PMID- 10676191 TI - Using WWW-based instruction modules and E-mail for a remote neurology course. PMID- 10676192 TI - Promoting reflective teaching with personal digital assistants. PMID- 10676193 TI - Web-based instruction to enhance the clinical teaching of community preceptors. PMID- 10676194 TI - Curriculum of literature and medicine for residents. PMID- 10676195 TI - A public-private collaboration to develop medical students' communication skills with patients. PMID- 10676196 TI - A new relationship module for doctor-patient encounters. PMID- 10676197 TI - Using patient narrative videos for understanding better the illness experience. PMID- 10676198 TI - Using standardized patients to teach end-of-life skills to clinical clerks. PMID- 10676199 TI - Teaching professionalism in medical grand rounds. PMID- 10676200 TI - Incorporating discussion of cultural diversity throughout the first-year medical curriculum. PMID- 10676201 TI - A comprehensive student peer-teaching program. PMID- 10676202 TI - Recruiting and following adolescent standardized patients. PMID- 10676203 TI - A four-year longitudinal gerontology curriculum for medical students. PMID- 10676204 TI - A critical care subinternship using the ICU as an applied physiology laboratory. PMID- 10676205 TI - Improving gross anatomy education by using surgery residents as co-teachers. PMID- 10676206 TI - A partnership in interdisciplinary clinical education. PMID- 10676207 TI - Reinvigorating PBL by integrating standardized-patient interviews. PMID- 10676208 TI - Implementing national guidelines for the internal medicine clerkship: an ambulatory case workbook. PMID- 10676209 TI - A diabetes self-care simulation for residents and medical students. PMID- 10676210 TI - Massachusetts Medical Society Seminar Series on Domestic Violence. PMID- 10676211 TI - An interactive, problem-based workshop on breastfeeding. PMID- 10676212 TI - Integrated clerkships for medical undergraduates. PMID- 10676213 TI - A clinical integration course. PMID- 10676214 TI - A geriatric medicine program in the internal medicine clerkship. PMID- 10676215 TI - A women's health course for education in internal medicine. PMID- 10676216 TI - The interdisciplinary case conference. PMID- 10676217 TI - Using respiratory therapists to teach arterial puncture for blood gas procedures to third-year medical students. PMID- 10676218 TI - The standardized family: a cross-institutional effort to standardize the ambulatory clerkship experience. PMID- 10676219 TI - Teaching medical documentation according to the HCFA guidelines. PMID- 10676220 TI - A tool to evaluate self-efficacy in evidence-based medicine. PMID- 10676221 TI - Comprehensive performance examination gives insights into the "hidden curriculum". PMID- 10676222 TI - Using telemedicine and standardized patients to evaluate off-campus students' skills. PMID- 10676223 TI - The brief structured observation--a tool for focused feedback. PMID- 10676224 TI - A first-year course focused on the patient, the physician, and the community. PMID- 10676225 TI - A longitudinal community clinical program for first-year medical students. PMID- 10676226 TI - A science learning initiative with urban junior high school students. PMID- 10676227 TI - Introducing home visits and interdisciplinary learning to an existing geriatrics practicum for medical students. PMID- 10676228 TI - Managed-care curriculum for family practice residents. PMID- 10676229 TI - Teaching managed-care principles to residents. PMID- 10676230 TI - A managed care curriculum implemented across four academic departments using mandated evaluation instruments. PMID- 10676231 TI - Scheduling solutions for the service-education conflict in internal medicine residency programs. PMID- 10676232 TI - A separate-sample pretest-post-test design to evaluate a practice-management seminar for residents. PMID- 10676233 TI - Training in behavioral medicine and behavioral change for family practice residents. PMID- 10676235 TI - A curriculum for pediatrics residents in development and evaluation of clinical practice guidelines. PMID- 10676234 TI - A new approach to the journal club. PMID- 10676236 TI - Medical education 101--a faculty development course. PMID- 10676237 TI - Development of a leadership-skills--assessment instrument for medical educators. PMID- 10676238 TI - A faculty development program in basic teaching skills. PMID- 10676239 TI - Faculty development using evidence-based medicine as an organizing curricular theme. PMID- 10676240 TI - Auscultation CME at the bedside for pediatrics practitioners. PMID- 10676241 TI - Improving communication in the referral-consultation process. PMID- 10676242 TI - Training opinion leaders to improve physician practice in the management of dyslipidemia. PMID- 10676243 TI - A curriculum for producing resident researchers. PMID- 10676244 TI - A four-month faculty development curriculum on teaching and learning. PMID- 10676245 TI - Using distance education for an MPH degree program in health services administration for physicians. PMID- 10676246 TI - Building administrative skills: a framework for junior faculty. PMID- 10676247 TI - Focus groups on curriculum and program evaluation. PMID- 10676248 TI - Center for Medical Education Research. PMID- 10676249 TI - Fourth-year students as teachers of the physical examination. PMID- 10676250 TI - Medical students as co-instructors in an introductory clinical medicine course. PMID- 10676251 TI - Faculty mentors for interaction over four years. PMID- 10676252 TI - A computer literacy requirement for medical students. PMID- 10676253 TI - Virtual PBL: full-scale human simulation technology. PMID- 10676254 TI - A multimedia and Internet program to present clinical cases. PMID- 10676255 TI - New sources for accessing and interpreting electronic information. PMID- 10676256 TI - Medical education over the Internet. PMID- 10676257 TI - Using computer technology for nutrition education and cancer prevention. PMID- 10676258 TI - A simulator-based respiratory physiology workshop. PMID- 10676259 TI - Combining service and learning in partnership with communities. PMID- 10676260 TI - A structured diary to promote reflection and active learning. PMID- 10676261 TI - Electronic learning contracts. PMID- 10676262 TI - Clinical instruction for delivering bad news. PMID- 10676263 TI - Introducing students to the spiritual dimension of illness. PMID- 10676264 TI - A student-run course in the medical humanities. PMID- 10676265 TI - A curriculum in medical record documentation. PMID- 10676266 TI - Addressing telephone medicine in the medical school curriculum. PMID- 10676267 TI - Health Policy Fellowship Programs. PMID- 10676268 TI - An educational conference to recruit women to primary care residencies. PMID- 10676269 TI - An experiential approach to alternative medicine. PMID- 10676270 TI - Breastfeeding demonstration in behavioral science curriculum. PMID- 10676271 TI - Integration of developmental psychology and preventive medicine. PMID- 10676272 TI - Linking basic, clinical, and social sciences. PMID- 10676273 TI - Total immersion for medical neuroscience. PMID- 10676274 TI - A generalist continuity experience in the community. PMID- 10676275 TI - Replacing lectures with reading, small-group discussion, and computer-assisted learning. PMID- 10676276 TI - Maximizing student-centered learning. PMID- 10676277 TI - GeriAction: a preclinical bedside course. PMID- 10676278 TI - The paper-case clinic: teaching medical physiology by PBL. PMID- 10676279 TI - Computer-based learning in PBL. PMID- 10676280 TI - A course in advanced clinical pharmacology and anesthesiology. PMID- 10676281 TI - Teaching occupational history-taking. PMID- 10676282 TI - Branching cases in problem-based learning. PMID- 10676283 TI - Using problem-based learning to target public health problems. PMID- 10676284 TI - Integrating women's health issues into the first-year curriculum. PMID- 10676285 TI - A multidisciplinary approach to a women's health curriculum. PMID- 10676286 TI - Implementation of a foundations in medicine course. PMID- 10676287 TI - The Vermont Generalist Curriculum. PMID- 10676288 TI - Longitudinal clinics for medical students. PMID- 10676289 TI - A palliative care clerkship for senior medical students. PMID- 10676290 TI - An interdisciplinary course on domestic and family violence. PMID- 10676291 TI - An evidence-based medicine seminar series. PMID- 10676292 TI - Teaching medical students about long-term illness. PMID- 10676293 TI - "Virtual delivery": an interactive role play of abnormal labor. PMID- 10676294 TI - Criterion-based evaluation in a third-year internal medicine clerkship. PMID- 10676295 TI - Group evaluation of student performance in a clerkship. PMID- 10676296 TI - Critique of a clinical curriculum from examination performance. PMID- 10676297 TI - An independent community-based ambulatory clerkship. PMID- 10676298 TI - A comprehensive multidisciplinary ambulatory clerkship. PMID- 10676299 TI - Development of a multidisciplinary ambulatory clerkship. PMID- 10676300 TI - Educating medical students in ambulatory clinics while maintaining patient flow. PMID- 10676301 TI - Interdisciplinary primary care summer preceptorships. PMID- 10676302 TI - Ethics curriculum for internal medicine residents. PMID- 10676303 TI - Psychosocial curriculum for a primary care residency. PMID- 10676304 TI - Psychiatric and psychosocial training for residents. PMID- 10676305 TI - Physician-patient communication in ambulatory settings. PMID- 10676306 TI - Joint training for residents and chaplain interns. PMID- 10676307 TI - Developing evaluation and review skills. PMID- 10676308 TI - A remedial course on interviewing skills. PMID- 10676309 TI - An industry-based practicum for occupational medicine. PMID- 10676310 TI - A rural model for GME. PMID- 10676311 TI - The "office-based medical team" for residents. PMID- 10676312 TI - Primary care training for ob-gyn practitioners. PMID- 10676313 TI - Distance learning in basic science for residents. PMID- 10676314 TI - A cost-effective program to give video/audiotape feedback. PMID- 10676315 TI - The IPX: early assessment for interns. PMID- 10676316 TI - On-going and college-wide faculty development. PMID- 10676317 TI - Improving the publication rate of fellowship research projects. PMID- 10676318 TI - Faculty development through distance-learning. PMID- 10676319 TI - Faculty development for community primary care preceptors. PMID- 10676320 TI - Global measure of teaching performance. PMID- 10676321 TI - Automating portfolio documentation for faculty. PMID- 10676322 TI - Building clinicians' teacher knowledge. PMID- 10676323 TI - Strategies to increase the number and the quality of innovations in Medical Education Grants (IMEGs). PMID- 10676324 TI - The student perception survey: a tool for assessing medical school curricula. PMID- 10676325 TI - Macro program evaluation: an approach to evaluating a generalist physician initiative program. PMID- 10676326 TI - Vital indicators of teaching and learning success (VITALS): stakeholders' perceptions of a course-improvement system. PMID- 10676327 TI - Student-run focus groups for the evaluation of pre-clinical courses. PMID- 10676328 TI - An early-acceptance program for underrepresented minority or economically disadvantaged students. PMID- 10676329 TI - An earlier approach to increase awareness of primary care careers. PMID- 10676330 TI - A psychiatry club as an extracurricular activity for medical students. PMID- 10676331 TI - A case-based integrated curriculum on the World Wide Web. PMID- 10676332 TI - Computer case simulations for student evaluation in a PBL track. PMID- 10676333 TI - LabSim: the western blot--a computer-based learning module. PMID- 10676334 TI - An Internet home page to log students' patient contacts. PMID- 10676335 TI - Clerkship evaluation by students: a standardized electronic mechanism. PMID- 10676336 TI - A Web-based evaluation system. PMID- 10676337 TI - Integrating basic science, clinical medicine, and applied research in an ambulatory clerkship. PMID- 10676338 TI - Teaching nutritional concepts by integrating basic science and introductory clinical courses. PMID- 10676339 TI - Using interdisciplinary educational strategies to improve the teaching of cellular neurodevelopment. PMID- 10676340 TI - Learning about the environment and health: an interclerkship experience for third year students. PMID- 10676341 TI - Integrating women's health issues into third-year medical and surgical curricula. PMID- 10676342 TI - An interclerkship course on domestic abuse. PMID- 10676343 TI - An integrated psychiatry-neurology clerkship within a problem-based learning curriculum. PMID- 10676344 TI - Integrating basic and clinical sciences in a course for senior medical and graduate students. PMID- 10676345 TI - The primary care continuum: basic science and clinical science integration in a classroom setting. PMID- 10676346 TI - Committing medical students to the ethics of medicine. PMID- 10676347 TI - Introducing and assessing bioethical training in an internal medicine clerkship. PMID- 10676348 TI - Integrating clinical ethical concepts and patient-centered problem solving into the basic science curriculum. PMID- 10676349 TI - A program to elucidate differences in medical students' communication skills. PMID- 10676350 TI - Incorporating multiculturalism into a doctor-patient course. PMID- 10676351 TI - Sequential assessment of medical student competence with respect to professional attitudes, values, and ethics. Subcommittee on Professional Attitudes and Values, Student Progress Assessment. PMID- 10676352 TI - A multi-modal assessment of behavioral competence. PMID- 10676353 TI - Teaching the sexual history: a tutorial approach. PMID- 10676354 TI - Teaching the development of children's concepts of illness. PMID- 10676355 TI - An extended evidence-based medicine curriculum for medical students. PMID- 10676356 TI - A medical technology assessment course. PMID- 10676357 TI - A preclinical practice skills program. PMID- 10676358 TI - Medical interviewing in Spanish. PMID- 10676359 TI - A family- and community-centered clinical curriculum. PMID- 10676360 TI - Comprehensive evaluation of an academic-community partnership program. PMID- 10676361 TI - A community-skills program in a preclinical curriculum. PMID- 10676362 TI - Shaping the third year of a primary care program in a tertiary care environment. PMID- 10676363 TI - A public health sub-curriculum of a pediatrics clerkship. PMID- 10676364 TI - Introducing students to issues of socioeconomic status and community services. PMID- 10676365 TI - Enhanced skill building. PMID- 10676366 TI - Medical student education: an admission and curricular approach to rural physician shortages. PMID- 10676367 TI - A year-long longitudinal third-year clerkship in an inner-city health center designed to maximize continuity. PMID- 10676368 TI - Introduction to health: a new discipline for the early exposure of medical students to public-health-related activities in Brazil. PMID- 10676369 TI - A teaching tool to enhance medical student education in ambulatory internal medicine. PMID- 10676370 TI - Using two-way interactive televideo for problem-based tutorials in a family medicine clerkship. PMID- 10676371 TI - Using standardized-patient instructors to teach students about the needs of patients with disabilities. PMID- 10676372 TI - Cardiovascular risk factor identification: an educational experience for third year medical students. PMID- 10676373 TI - Removing rater effects from medical clerkship evaluations. PMID- 10676374 TI - An alternative to clinical practice examinations. PMID- 10676375 TI - Improving feedback with a clinical encounter form. PMID- 10676376 TI - Hidden assets: incorporating the department business manager into training. PMID- 10676377 TI - A practice-management experience in rural practice. PMID- 10676378 TI - Practice-management curriculum for family practice residents. PMID- 10676379 TI - Teaching residents in an ambulatory setting: introducing the concepts of managed care in a fee-for-service clinic. PMID- 10676380 TI - Training in community service and public health for internal medicine residents. PMID- 10676381 TI - A comprehensive community-based program for training internal medicine residents in ambulatory settings. PMID- 10676382 TI - A nursing home experience for internal medicine residents. PMID- 10676383 TI - A graduate medical experience in interdisciplinary group learning. PMID- 10676384 TI - Incorporating internal medicine residents into an interdisciplinary geriatric assessment team. PMID- 10676385 TI - Expanding the pediatrics residency curriculum. PMID- 10676386 TI - A sports medicine curriculum for pediatrics residents. PMID- 10676387 TI - Development of a breastfeeding curriculum. PMID- 10676388 TI - An instructional module in musculoskeletal examination for residents incorporating physical therapists as patient-instructors and evaluators. PMID- 10676389 TI - Using simulated students to improve residents' teaching. PMID- 10676390 TI - Evaluating an evidence-based medicine curriculum. PMID- 10676391 TI - Facilitated behavior-based evaluation of resident performances on an inpatient rotation. PMID- 10676392 TI - Analyzing the questions of physicians participating in CME programs. PMID- 10676393 TI - An integrated approach to CME and service needs identification, program development, and information diffusion. PMID- 10676394 TI - Evaluating continuing competence of physicians through multiple assessment modalities: the Physicians' Continued Competence Assessment Program (PCCAP). PMID- 10676395 TI - An examination of the feasibility of developing and offering courses that meet MAINPRO-C requirements. PMID- 10676396 TI - Scholars for teaching excellence: institutionalizing faculty development. PMID- 10676397 TI - A systems-based approach to improving educational quality via community-oriented faculty development. PMID- 10676398 TI - Improving written narrative assessments in small-group, problem-based tutorials: continuous quality assurance and faculty development through peer review. PMID- 10676399 TI - Enhancing awareness of diversity and cultural competence: a workshop series for department chairs and course directors. PMID- 10676400 TI - A situational approach to improve clinical teaching. PMID- 10676401 TI - A workshop to improve faculty members' administrative skills. PMID- 10676402 TI - A multidisciplinary experiential course in instructional methodology for academic clinicians. PMID- 10676403 TI - An evidence-based education journal club. PMID- 10676404 TI - Building faculty skills as educators: a total quality management approach. PMID- 10676405 TI - Risk assessment for children and other sensitive populations. AB - Children form a unique subgroup within the population who require special consideration in risk assessment. Children are not little adults. Their tissues and organs grow rapidly, developing and differentiating. These development processes create windows of great vulnerability to environmental toxicants. Furthermore, the exposure patterns of children to environmental chemicals are very different from those of adults. Traditional risk assessment has generally failed to consider the special exposures and the unique susceptibilities of infants and children. Adoption of a new child-centered agenda for research and risk assessment is necessary if disease in children of toxic environmental origin is to be identified, understood, controlled, and prevented. This agenda needs to be multidisciplinary. Specific requirements within the agenda include: (1) exploration and quantification of unique patterns of exposure for children; (2) adoption of new, more sensitive approaches to testing chemicals that can recognize the consequences of exposure during early development; (3) identification, through clinical and epidemiologic studies, of etiologic associations between environmental exposures and pediatric diseases; and (4) elucidation, at the cellular and molecular levels, of the pathogenetic mechanisms of pediatric environmental illness. In the United States, an important start toward adoption of this new agenda has occurred since passage of the Food Quality Protection Act in 1996. A Presidential Executive Order on Children's Health and the Environment has been promulgated. This Order requires all federal agencies to make protecting the health of children against environmental hazards a high priority. A new Office of Children's Health Protection has been established at the U.S. Environmental Protection Agency. Programs in children's environmental health have been created at the Centers for Disease Control and Prevention, the Agency for Toxic Substances and Disease Registry, and the National Institute of Environmental Health Sciences. A national network of eight new Children's Environmental Health Research and Disease Prevention Centers has been formed. These developments will enhance research on previously understudied issues in the environmental health of children and will provide a scientific basis for child centered risk assessment. PMID- 10676406 TI - The scientific and methodological bases of experimental studies for detecting and quantifying carcinogenic risks. AB - This paper outlines the aims and potential scope of experimental research for risk identification and assessment in industrial carcinogenesis (environmental and occupational). It then reviews the basic, general, and specific requisites of a rigorously scientific nature that are required to render experiments to be more appropriate and better geared to the information they seek. A range of experimental approaches to risk assessment are illustrated by results achieved in the Cancer Research Centre of the Ramazzini Foundation (CRC/RF). The paper ends with a call for closer relations and integration among experimental, epidemiologic, and biostatistical studies. PMID- 10676407 TI - A Bayesian approach to hazard identification. The case of electromagnetic fields and cancer. AB - This paper discusses certain issues related to uncertainty in hazard identification. Research on the hypothesis that exposure to 50-60-Hz magnetic and electric fields (EMF) increases the risk of cancer has been ongoing for two decades. Epidemiological studies provide a somewhat consistent pattern indicating an increased risk for childhood leukemia and adult chronic lymphatic leukemia and possibly also for other leukemias and brain cancer. However, there is still no good candidate for a mechanism. Epidemiological studies have throughout the two decades been interpreted with great caution, and final evaluations as to carcinogenicity have been deferred. The reason for this carefulness may be the lack of knowledge about a plausible mechanism. The purpose of this paper is to discuss the process of weighing epidemiological data, experimental data, and other background information into a synthesis such that the evaluation can be based on all data combined. A Bayesian approach to this weighing is discussed along with some alternatives. The Bayesian approach provides a structure for the pooling of evidence and points out where subjective judgments come into play. PMID- 10676408 TI - Mega-experiments to identify and assess diffuse carcinogenic risks. AB - Diffuse carcinogenic risks, that is, those of low potency involving large areas of population and sometimes all mankind, pose a serious public health problem. Controlling these risks might help to reduce the incidence of, and mortality from, cancer. Because of their low expected carcinogenic potential, these risks are difficult to expose or assess. Epidemiologic investigation is of limited use in this field and yields its data too late to be useful. Experimental studies offer the only possible approach for assessing such risks. To increase experimental sensitivity and consistency of results, mega-experiments must be designed. That is, experiments that use a large number of animals with a well known basic tumorigram, that extend the exposure and the biophase for as long as possible, that carefully observe the effects, and that are performed with suitable standardized methods. In the last 15 years the Ramazzini Foundation, in its Cancer Research Center at Bentivoglio, has conducted or planned five mega experiments. Initial results indicate the great potential of these methods for identifying and assessing diffuse risks. PMID- 10676409 TI - Long-term chemical carcinogenesis bioassays predict human cancer hazards. Issues, controversies, and uncertainties. AB - Long-term carcinogenesis bioassays are the most valued and predictive means for identifying potential carcinogenic hazards of various agents to humans. Agents may be chemicals, chemical mixtures, multiple chemicals, combinations of chemicals, residues and contaminants, commercial products and formulations, and various exposure circumstances. Life-styles, dietary factors, and occupational exposure circumstances are very difficult, but not totally impossible, to evaluate experimentally. Historically, the first chemical bioassay took place in the early part of this century: Yamagiwa and Ichikawa in 1915, showed that coal tar applied experimentally to rabbit ears caused skin carcinomas. Since then, nearly 1500-2000 bioassays of one sort or another have been carried out. Importantly, however, some of these bioassays must be considered inadequate for judging the absence of carcinogenicity, since there were various limitations on the way they were performed: too few animals, too short a duration, too low exposure concentrations, too limited pathology, as examples. Thus, each bioassay must be critically evaluated, especially those reported to be negative, because "false negatives" are certainly more hazardous to human health than are "false positives". Likewise, one must be careful not to discount bioassay results simply because a target organ in rodents may not have a direct counterpart in humans (e.g., Zymbal glands), or because an organ site in rodents may not be a major site of cancers in humans (e.g., mouse liver). The design and conduct of a bioassay is not simple, however, and one must be fully aware of possible pitfalls as well as viable and often necessary alternatives. Similarly, evaluating results and interpreting findings must be approached with the utmost objectivity and consistency. These and other select issues, controversies, and uncertainties possibly encountered in long-term bioassays are covered in this paper. One fact remains abundantly clear: for every known human carcinogen that has been tested adequately in laboratory animals, the findings of carcinogenicity are concordant. PMID- 10676410 TI - Uncertainty in biomonitoring and kinetic modeling. AB - Uncertainty in exposure assessment and uncertainty in kinetic models of early effects after exposure to a toxin are addressed in this paper. Sources of uncertainty in the determination of exposure of workers in chemical industry exposed to dioxins are exhibited and a simple kinetic model for biomonitor measurements of the concentrations from occupational exposure is derived. Model uncertainty, and uncertainty in the model parameters of physiologically-based pharmacokinetic models (PBPK models) are addressed when these models are used to estimate the effective dose in risk assessment. Uncertainty in the model parameters originating from the use of different statistical analysis methods is exhibited for Hill type nonlinear kinetics of enzyme induction mediated by a toxin. PMID- 10676411 TI - Using molecular epidemiology in assessing exposure for risk assessment. AB - Quantitative estimation of health risks depends on exposure characterization, the nature of the dose response relationships, and the toxicity of the agents involved. The greatest uncertainties in risk assessment almost always arise from sparse or inadequate exposure data, inadequate understanding of exposure mechanisms, and insufficient understanding of the exposure-dose-response pathway. Additional sources of uncertainty arise when mixed or multiple exposures are implicated in the disease pathway, and as a result of variability in both exposures and responses within and between individuals. Here we consider the role of exposure assessment in the risk assessment process, the use of biological markers or molecular epidemiology to contribute to improvements in exposure assessment for risk assessment, and uncertainties associated with the use of biological markers. PMID- 10676412 TI - Kaplan-Meier tumor probability as a starting point for dose-response modeling provides accurate lifetime risk estimates from rodent carcinogenicity studies. AB - In rodent carcinogenicity studies the linearized multistage model for modelling the dose-response for specific tumor incidence has limitations in accuracy. This note provides an alternative basic method for analyzing the dose-response relationship. It is based on an actuarial analysis of mortality and specific tumor incidence. The survival and the Kaplan-Meier specific tumor probability are fitted to a Weibull model, in which exposure level, exposure period, and observation period are independent variables. The mortality from specific cancers at a certain time is simulated by means of the product of survival and specific tumor rate (derivative of Kaplan-Meier tumor probability) as function of exposure level, exposure duration, and observation period, integrated over the observation period. The model is demonstrated by means of fitting the mortality and tumor incidence data from the second NTP mice study on butadiene to a Weibull model and to the linear, so-called, one-hit model. It will be shown that, in the experimental exposure range, the Weibull model is far superior to the one-hit model and predicts the specific tumor incidence with a high accuracy over the total dose range. The Kaplan-Meier probability model for a specific tumor is also useful for regulatory risk estimation. It is proposed that to develop a specific tumor a risk level of 1 in 1,000 over a lifetime is about equal to 5 in 10,000 at 50% survival of the population. The Kaplan-Meier probability may be estimated at the time of 50% survival of the exposed population, which can be deduced from the all mortality data. This estimation method provides meaningful data, using exposure level, exposure duration, and observation period properly. The advantage of the actuarial analysis method for interpreting rodent studies is that allowance is made for competition between death causes, which is essential in case of considerable difference in mortality and specific mortality between dose groups. Integrating the product of survival and specific tumor rate is the proper way to predict, comparatively, mortality and specific mortality in exposed and unexposed rodent populations. PMID- 10676413 TI - Uncertainty in estimating exposure using a toxicokinetic model. The example of 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - This paper deals with sources of uncertainty in the use of a minimal physiological toxicokinetic model to obtain dose estimates for a dose-response analysis of cancer in an occupational cohort. Toxicokinetic models make it possible to construct exposure parameters that are more closely related to the individual dose than traditional measures of exposures to toxic agents. However, the process introduces a wide array of sources of uncertainty. Selecting a model structure to describe the kinetics of a toxic agent implies necessarily making simplifications and assumptions that influence the range of applicability of the model. Once a model has been selected, the value of certain model parameters (constants) must be assigned, for example, from anthropometric data. The question then arises of how sensitive the model predictions are to variations in the values of these constants. Other model parameters, typically those describing the kinetics of the agent, are next estimated from actual data. There may be limitations in the data concerning, for example, sparseness (too few observations per subject) or missing values. The methods used for parameter estimation carry their own set of assumptions that need to be appropriate to the situation at hand. In summary, the dioxin example is used to characterize the sources of uncertainty at different levels, such as model structure, methods and data used for parameter estimation, estimation of occupational exposure, and imputation of missing values in exposure indices derived from the kinetic model. PMID- 10676414 TI - Uncertainty in the relation between exposure to magnetic fields and brain cancer due to assessment and assignment of exposure and analytical methods in dose response modeling. AB - Incomplete scientific knowledge ensures that, in every study, uncertainty will enter the processes of exposure estimation and exposure-response modeling. In the light of the heated debate about the health effects of magnetic fields resulting from power production and usage, we undertook a sensitivity analysis to evaluate uncertainty related to key decisions in a previous study of brain cancer and occupational exposure to magnetic fields. The findings appeared to be relatively insensitive to most variations in the methods of exposure assessment, exposure assignment, and data analysis. The results can be visualized by defining bands of uncertainty about a best-bet estimate of the association based on our original study. These bands of methodological uncertainties were similar in magnitude to the conventional 95% confidence interval, but they provide a measure of the potential range of systematic bias in the results, rather than reflecting statistical variability alone. The methodology employed here can be applied to other studies, and other researchers are encouraged to conduct sensitivity analysis in order to estimate methodological uncertainty as an alternative to statistical confidence intervals. PMID- 10676415 TI - Measures of exposure to environmental tobacco smoke. Validity, precision, and relevance. AB - It is often not clear what the best measures of exposure are for a risk assessment, or even how one should answer this question. Environmental tobacco smoke (ETS) provides a good example for an exploration of uncertainty. There are a variety of methods for estimating exposure and each has short-comings. In this paper we summarize the physical characteristics of ETS and the principal methods for assessing exposure. We review the accuracy and applicability of these methods, and explore major sources of uncertainty in the assessment of ETS. PMID- 10676416 TI - The contribution of environmental monitoring in the epidemiological assessment of exogenous risk. The experience of ARPA in the Emilia-Romagna Region of Italy. AB - The aim of the Emilia Romagna-Region Agency for Prevention and Environment (ARPA) is to define and improve interactions among the various prevention departments of the Emilia-Romagna Local Health Authorities in order to attain better knowledge about the health status of the population by using epidemiology and etiology studies, as well as predictive models. This is the basis for the environmental health risk assessment strategy of ARPA. The priority activity areas for ARPA are: urban areas, environmental and health effects of traffic (atmospheric pollution and noise pollution); industrial areas (Ravenna chemical plants, Modena/Reggio-Emilia ceramic factories and Ferrara chemical plants); high-speed trains; pesticides; asbestos; and pollution of the Adriatic Sea. PMID- 10676417 TI - Combining uncertainty factors in deriving human exposure levels of noncarcinogenic toxicants. AB - Acceptable levels of human exposure to noncarcinogenic toxicants in environmental and occupational settings generally are derived by reducing experimental no observed-adverse-effect levels (NOAELs) or benchmark doses (BDs) by a product of uncertainty factors (Barnes and Dourson, Ref. 1). These factors are presumed to ensure safety by accounting for uncertainty in dose extrapolation, uncertainty in duration extrapolation, differential sensitivity between humans and animals, and differential sensitivity among humans. The common default value for each uncertainty factor is 10. This paper shows how estimates of means and standard deviations of the approximately log-normal distributions of individual uncertainty factors can be used to estimate percentiles of the distribution of the product of uncertainty factors. An appropriately selected upper percentile, for example, 95th or 99th, of the distribution of the product can be used as a combined uncertainty factor to replace the conventional product of default factors. PMID- 10676418 TI - Statistical methods for developmental toxicity. Analysis of clustered multivariate binary data. AB - This paper discusses some of the statistical issues that arise from developmental toxicity studies, wherein pregnant mice are exposed to chemicals in order to assess possible adverse effects on developing fetuses. We begin with a review of some current approaches to risk assessment, based on NOAELs, and provide justification for the use of methods based on dose-response models. Due to the hierarchical nature of the data, such models are more complicated in the present context than, say, in cancer studies. For example, multivariate binary outcomes arise when each fetus in a litter is assessed for the presence of malformations and/or low birth weight. We describe a multivariate exponential family model that works well for these data and that is flexible in terms of allowing response rates to depend on cluster side. Maximum likelihood estimation of model parameters and the construction of score tests for dose effect are briefly discussed. Results are illustrated with data from several NTP studies. PMID- 10676419 TI - Sources of uncertainty in dose-response modeling of epidemiological data for cancer risk assessment. AB - Epidemiologic data is increasingly being used for dose-response analysis in risk assessment. The Environmental Protection Agency (EPA) and other U.S. agencies have expressed a preference for using epidemiologic data rather than toxicologic data when possible. However, there are a number of important sources of uncertainty in using epidemiologic data for this purpose that need to be clearly recognized and, when possible, quantified. This paper presents a critical review of the major sources of uncertainty in the use of epidemiologic data for cancer risk assessment. These may include: (1) study design issues such as potential confounding and other biases, inadequate sample size, and followup, (2) the choice of the data set, (3) specification of the dose-response model, (4) estimation of exposure and dose, and (5) unrecognized variability in susceptibility. Examples from risk assessments for cadmium, asbestos, and diesel exhaust are used to illustrate the potential magnitude of some of these sources of uncertainty. It is shown that the overall uncertainty from these various sources combined may often result in highly uncertain risk estimates from dose response modeling of epidemiologic data. For this reason, we believe it is best to present a range of possible risk estimates, which, to the extent possible, reflects the variability and uncertainty inherent in the dose-response evaluation of epidemiologic data. PMID- 10676420 TI - Nonparametric analysis of dose-response relationships. AB - A nonparametric method, isotonic regression, is proposed for analyzing a dose response relationship and for assessing a threshold value. There are several advantages of this method compared to parametric models. No specific form of the relationship (type of model and use of the covariates) is required. The only assumption is monotonicity. Rejection of specific hypothesis can be based on the result of a permutation test. Several applications (para-aramid, crystalline silica, and PNOC) are presented. In these examples the dose-response relationships are analyzed. Where a relationship is present the existence of a threshold is investigated. PMID- 10676421 TI - Estimates of the proportions of carcinogens and anticarcinogens in bioassays conducted by the U.S. National Toxicology Program. Application of a new meta analytic approach. AB - A meta-analysis was performed in order to estimate the proportion of liver carcinogens, the proportion of chemicals carcinogenic at any site, and the corresponding proportion of anticarcinogens among chemicals tested in 397 long term cancer bioassays conducted by the U.S. National Toxicology Program (NTP). Although the estimator used was negatively biased, the study provided persuasive evidence for a larger proportion of liver carcinogens (0.43, 90% CI: 0.35, 0.51) than was identified by the NTP (0.28). A larger proportion of chemicals carcinogenic at any site was also estimated (0.59, 90% CI: 0.49, 0.69) than was identified by the NTP (0.51), although this excess was not statistically significant. A larger proportion of anticarcinogens (0.66) was estimated than carcinogens (0.59). Despite the negative bias, it was estimated that 85% of the chemicals were either carcinogenic or anticarcinogenic at some site in some sex species group. This suggests that most chemicals tested at high enough doses will cause some sort of perturbation in tumor rates. PMID- 10676422 TI - Characterization of uncertainty and variability in residential radon cancer risks. AB - Radon, a naturally occurring gas found at some level in most homes, is an established risk factor for human lung cancer. The U.S. National Research Council has recently completed a comprehensive evaluation of the health risks of residential exposure to radon and developed models for projecting radon lung cancer risks to the general population. This analysis suggests that radon may play a role in the etiology of 10-15% of all lung cancer cases in the United States, although these estimates are subject to considerable uncertainty. In this article, we present a detailed analysis of uncertainty and variability in estimates of lung cancer risk due to residential exposure to radon. We use a general framework for the analysis of uncertainty and variability that we developed previously. Specifically, we focus on estimates of the age-specific excess relative risk (ERR) and lifetime relative risk (LRR), both of which vary substantially among individuals. We also consider estimates of the population attributable risk (PAR), which reflects the proportion of the lung cancer burden attributable to radon. Variability in the ERR and LRR is largely determined by variability in residential exposure levels and in the dosimetric K-factor used to extrapolate from occupational to environmental settings. Uncertainty in the ERR and LRR is due to uncertainty in the model parameters, notably those reflecting the carcinogenic potency of radon and the modifying effect of attained age. Uncertainty in the PAR is determined by uncertainty about the values of the parameters in the risk models used to estimate the PAR. Uncertainty in radon levels in homes and the dosimetric K-factor contribute comparatively little to uncertainty in the PAR. These results suggest that reduction in uncertainty about the PAR for radon induced lung cancer can only be achieved if more reliable risk projection models can be developed. PMID- 10676423 TI - Risk assessment--the mother of all uncertainties. Disciplinary perspectives on uncertainty in risk assessment. AB - Uncertainty in the detection and evaluation of chemical hazards to health leads to challenges when conducting risk assessments. Some of the uncertainty has to do with data, some with incomplete understanding of processes, and some with the most fundamental ways of viewing the questions. True variability--across space, in time, or among individuals--complicates the search for understanding many important aspects of risk. A few statistical and toxicologic tools are available to assess uncertainty. Three methods of classifying uncertainty are briefly discussed. In addition, our disciplinary background may influence how we view and discuss variability and uncertainty. We rarely know as much as we think we do (and not just in risk assessment). Great uncertainty is likely to remain an important part of risk assessment for some decades to come. PMID- 10676424 TI - Distributions of individual susceptibility among humans for toxic effects. How much protection does the traditional tenfold factor provide for what fraction of which kinds of chemicals and effects? AB - A significant data base has been assembled on human variability in parameters representing a series of steps in the pathway from external exposure to the production of biological responses: contact rate (e.g., breathing rates/body weight, fish consumption/body weight); uptake or absorption (mg/kg)/intake or contact rate; general systemic availability net of first pass elimination and dilution; systemic elimination or half-life; active site availability/general systemic availability; physiological parameter change/active site availability; functional reserve capacity--change in baseline physiological parameter needed to pass a criterion of abnormal function or exhibit a response. This paper discusses the current results of analyzing these data to derive estimates for distributions of human susceptibility to different routes of exposure and types of adverse effects. The degree of protection is tentatively evaluated by projecting the incidences of effects that would be expected for a tenfold lowering of exposure from a 5% incidence level if the population distribution of susceptibility were truly log-normal out to the extreme tails, and if the populations, chemicals, and responses that gave rise to the underlying data were representative of the cases to which traditional uncertainty factor is applied. The results indicate that, acting by itself, a tenfold reduction in dose from a 5% effect level is associated with effect incidences ranging from slightly less than one in ten thousand, for a median chemical/response, to a few per thousand, for chemicals and responses that have greater human interindividual variability than 19 out of 20 typical chemicals/responses. In practice, for many of the cases where the traditional tenfold factor is applied, additional protection is provided by other uncertainty factors. Nevertheless, the results generate some reason for concern that current application of traditional safety or uncertainty factor approaches may allow appreciable incidences of responses in some cases. PMID- 10676425 TI - Analysis of PBPK models for risk characterization. AB - Adoption of a Bayesian framework for risk characterization permits the seamless integration of different kinds of information available in order to choose and parameterize risk models. It also becomes easy to disentangle uncertainty from variability, through hierarchical statistical modeling. Appropriate numerical techniques can be found, for example, in the recently developed arsenal of Markov chain, Monte Carlo simulations. The developments in this area can actually be viewed as extensions of the traditional or standard Monte Carlo methods for uncertainty analysis. Following a brief review of the techniques, examples of Bayesian analyses of physiologically-based pharmacokinetic models are presented for tetrachloroethylene and dichloromethane. The discussion touches on some open problems and perspectives for the proposed methods. PMID- 10676426 TI - Uncertainty in risk characterization of weak carcinogens. AB - Epidemiologic inference is subject to uncertainty that is inherent to observational approaches. It was shown by the present analysis that an SMR of 1.4, as observed for 2,3,7,8-tetrachlorodibenzo-para-dioxin in the pooled analysis by IARC, would be observed in the absence of carcinogenic risk, if the smoking prevalence of cohort members was as low as 80%. It was also shown that a 1.4-fold increase in lung cancer risk among nonsmokers, which roughly corresponds to a daily consumption of one cigarette, might be extremely difficult to identify if the subjects are exposed to strong carcinogens such as those that result from cigarette smoking. On the other hand, a genotoxic agent that increases the lung cancer risk of smokers 1.4-fold, could increase the risk to nonsmokers by as much as 6.3-fold. In light of these uncertainties in epidemiologic inference, attempts to estimate the absolute risk in human populations by epidemiologic studies should be made with caution. PMID- 10676427 TI - Reducing uncertainty in the derivation and application of health guidance values in public health practice. Dioxin as a case study. AB - We were requested by the U.S. Environmental Protection Agency (EPA) to clarify the relationships among the minimal risk level (MRL), action level, and environmental media evaluation guide (EMEG) for dioxin established by the Agency for Toxic Substances and Disease Registry (ATSDR). In response we developed a document entitled "Dioxin and Dioxin-Like Compounds in Soil, Part I: ATSDR Interim Policy Guideline"; and a supporting document entitled "Dioxin and Dioxin Like Compounds in Soil, Part II: Technical Support Document". In these documents, we evaluated the key assumptions underlying the development and use of the ATSDR action level, MRL, and EMEG for dioxin. We described the chronology of events outlining these different health guidance values for dioxin and identified the areas of uncertainty surrounding these values. Four scientific assumptions were found to have had a great impact on this process; these were: (1) the specific uncertainty factors used, (2) the toxicity equivalent (TEQ) approach, (3) the fractional exposure from different pathways, and (4) the use of body burdens in the absence of exposure data. This information was subsequently used to develop a framework for reducing the uncertainties in public health risk assessment associated with exposure to other chemical contaminants in the environment. Within this framework are a number of future directions for reducing uncertainty, including physiologically based pharmacokinetic modeling (PBPK), benchmark dose modeling (BMD), functional toxicology, and the assessment of chemical mixture interactions. PMID- 10676428 TI - Uncertainty in risk characterization and communication. Discussion. PMID- 10676429 TI - Uncertainty in risk assessment. Current efforts and future hopes. AB - The incorporation of sampling variability in estimates of excess risk has been part of risk assessment practice for decades. Currently, there is a strong desire to incorporate understanding of biological mechanisms into the models used for exposure assessment and exposure-response modeling. In addition, representing population heterogeneity in the assessment of risks and the identification of sensitive subpopulations is of great concern. Finally, the communication of uncertainty and variability remains a challenge to risk assessors. Based upon the presentations of workshop faculty, a summary of current practice when addressing uncertainty together with conjectures concerning future challenges for addressing uncertainty, are presented. PMID- 10676430 TI - Common themes at the workshop on uncertainty in the risk assessment of environmental and occupational hazards. PMID- 10676431 TI - New vistas for melanocortins. Finally, an explanation for their pleiotropic functions. AB - This paper presents a historical overview of melanocortin (MC) research from the early investigations of the many noncorticotropic effects of peptide fragments of adrenocorticotropic hormone to the present focus on the discovery and cloning of the MC receptors (MCRs). Final acceptance of the passage of neuropeptides through the blood-brain barrier provided the scientific basis for the neuropeptide concept, formulated previously by both De Weid and Kastin, that peripherally administered neuropeptides affect neural processes. The discussion includes melanocortin effects on behavior, the cardiovascular system, central and peripheral electrophysiological parameters, food intake, inflammation and analgesia, nerve regeneration and neuroprotection, and development. The localization of specific MCRs in both neural and nonneural tissues is correlated with the pleiotropic effects discussed. PMID- 10676432 TI - Vasoactive intestinal peptide. Link between electrical activity and glia-mediated neurotrophism. AB - Vasoactive intestinal peptide has neurotrophic and neuroprotective properties that influence the survival of activity-dependent neurons in the central nervous system. Investigations of the mechanism of this neurotrophic peptide indicated that these actions are contingent on interactions with astroglia. The complex mixture of neurotrophic mediators released from astroglia include cytokines, a protease inhibitor, and activity-dependent neurotrophic factor, a protein with apparent structural similarities to hsp60. Investigations of ADNF resulted in the discovery of active peptides of extraordinary potency and broad neuroprotective properties. These studies indicate that a nine-amino acid core peptide of ADNF had significantly greater neuroprotective properties in comparison to the parent growth factor and these advantages identify ADNF-9 as an attractive lead compound for drug development. PMID- 10676433 TI - Stress, corticotropin-releasing factor, and drug addiction. AB - The neuropeptide corticotropin-releasing factor (CRE) and related neuropeptides not only mediate hormonal responses to stressors but also have a neurotropic role in the central nervous system to mediate behavioral responses to stressors. CRF antagonists effectively block CRF responses and have effects opposite those of CRF in many stress-related situations. Recent advances suggest that in addition to CRF itself there is another CRF-related neuropeptide, urocortin, that may be involved in stress-related responses, particularly those involving appetite. At least two CRF receptors have been discovered to date, CRF-1 and CRF-2. CRF may be involved in various aspects of the addiction cycle associated with drugs of abuse. CRF appears to be activated during stress-induced reinstatement of drug taking as well as acute withdrawal from all major drugs of abuse. CRF is hypothesized to be part of an allostatic change leading to vulnerability to relapse during prolonged abstinence from drugs of abuse. PMID- 10676434 TI - Evolution of the corticotropin-releasing hormone signaling system and its role in stress-induced phenotypic plasticity. AB - Developing animals respond to variation in their habitats by altering their rates of development and/or their morphologies (i.e., they exhibit phenotypic plasticity). In vertebrates, one mechanism by which plasticity is expressed is through activation of the neuroendocrine system, which transduces environmental information into a physiological response. Recent findings of ours with amphibians and of others with mammals show that the primary vertebrate stress neuropeptide, corticotropin-releasing hormone (CRH), is essential for adaptive developmental responses to environmental stress. For instance, CRH-dependent mechanisms cause accelerated metamorphosis in response to pond-drying in some amphibian species, and intrauterine fetal stress syndromes in humans precipitate preterm birth. CRH may be a phylogenetically ancient developmental signaling molecule that allows developing organisms to escape deleterious changes in their larval/fetal habitat. The response to CRH is mediated by at least two different receptor subtypes and may also be modulated by a secreted binding protein. PMID- 10676435 TI - Corticotrophin-releasing hormone and CRH-binding protein. Differences between patients at risk for preterm birth and hypertension. AB - BACKGROUND: During pregnancy in the second and third trimester there is a progressive rise in plasma CRH thought to be secreted by the placenta. Plasma CRH BP inactivates CRH, which may prevent its peripheral action on the maternal pituitary and myometrium. In the last few weeks of pregnancy CRH-BP decreases, thereby causing an increase in free CRH or a CRH/CRH-BP complex available to play a role in the onset of parturition. OBJECTIVE: We tested the hypothesis that differences in CRH, CRH-BP, or a CRH/CRH-BP complex in patients at risk for preterm birth (PTB) and hypertension (HYP) account for the differences in the timing of parturition. METHODS: From a Behavior in Pregnancy Study database, we identified 18 patients who had spontaneous PTB and 23 patients who developed HYP. Both groups were case controlled and matched with patients who delivered at term (Normal). Maternal plasma samples had been appropriately collected from these patients at 18-20, 28-30, and 35-36 weeks gestational age. CRH levels were measured by double antibody RIA kit and the CRH-BP by a immunoradiometric technique. A CRH-BP/CRH dimer complex index was calculated. Statistical analysis was done using Kruskal-Wallis test for two cases. RESULTS: Maternal CRH (pg/ml) in the PTB cases compared to the HYP cases was significantly elevated at all three time periods. Maternal CRH-BP (pg/ml) in the PTB versus HYP cases was significantly lower at all three time periods in the PTB cases compared to the HYP cases. Maternal CRH-BP/CRH dimer complex index was significantly lower in the PTB cases at all three time periods than either the controls or the HYP cases, suggesting excessive CRH. The mean GA at delivery for the PTB cases was significantly lower than the control or HYP cases. CONCLUSIONS: These results suggest that those patients at risk for PTB have significantly elevated CRH, lower CRH-BP, and a reduced CRH-BP/CRH dimer complex index at all three time periods of assessment. PMID- 10676436 TI - Corticotrophin-releasing hormone and fetal responses in human pregnancy. AB - During human pregnancy, maternal and fetal compartments of the human placenta produce and release corticotrophic-releasing hormone (CRH). Elevations of placental CRH are associated with decreased gestational length (including preterm delivery). The effects of elevated placental CRH on human fetal neurological development are not known. Pregnant women in the 31st and 32nd week of gestation consented to procedures for collection of blood and measurement of fetal heart rate (FHR) in response to a series of 40 vibro-acoustic stimuli (VAS). Measures of habituation and dishabituation were calculated from the FHR. All subjects were followed to delivery. Fetuses (N = 33) of women with highly elevated CRH were least responsive (p < .03) to stimulation after presentation of a novel (dishabituating) stimulus with control for parity, fetal gender, medical (antepartum) risk, and gestational length at term. In a larger sample (N = 156) a polynomial model predicted the pattern of FHR reactivity for the first 15 trials. Placental CRH concentration significantly predicted FHR reactivity after controlling for the effects of trial number, baseline FHR, inter-trial interval, and presence of uterine contractions. Increased maternal CRH levels were significantly related to the length of gestation after controlling for the effects of fetal gender, parity, and medical risk (p = .05). The relationship between length of gestation and FHR was not significant suggesting separate actions of CRH on these events. Elevated placental CRH appears to accelerate certain developmental events (gestational length) and may influence the fetal nervous system. The impaired fetal responses to novelty and increased arousal observed in this study suggest that neurological systems may be targets for placental CRH during sensitive developmental periods. PMID- 10676437 TI - Short- and long-term consequences of corticotropin-releasing factor in early development. AB - Corticotropin-releasing factor (CRF) mediates various stress-related responses in adult animals. Little is known about the effects of CRF during early development. Young mammals often vocalize when isolated in novel surroundings. Heightened levels of CRF inhibit vocalizing in isolated rat and guinea pig pups. Still lower levels of CRF may facilitate or permit vocalizing in rat pups. In guinea pigs, CRF appears to move pups from an initial active, to a subsequent passive, stage of behavioral responsiveness. CRF activity prior to birth can also affect the young. Exposing pregnant female rats to stressors during the last trimester of pregnancy alters the morphological and behavioral development of the offspring. Effects of gestational stress can be mimicked by injecting pregnant females with CRF during the last trimester. CRF appears to mediate both short- and long-term responses to stressors during developmental in rodents. PMID- 10676438 TI - Vasoactive intestinal peptide regulates embryonic growth through the action of activity-dependent neurotrophic factor. AB - Activity-dependent neurotrophic factor is a potent, neuroprotective protein released from astroglia by VIP and accounts in part for the neuroprotective properties of this neuropeptide. The growth-regulatory actions of VIP during embryogenesis may also occur indirectly through the release of activity-dependent neurotrophic factor. Whole cultured day-9 mouse embryos treated with activity dependent neurotrophic factor (10(-13) M) for 4 hr grew 3.1 somites, compared with 1.6 somites in control embryos. Treated embryos appeared morphologically normal and exhibited significant increases in cross-sectional area, protein, and DNA content and bromodeoxyuridine incorporation. Anti-activity-dependent neurotrophic factor significantly inhibited growth. Co-treatment of embryos with anti-activity-dependent neurotrophic factor inhibited VIP-stimulated growth; however, anti-VIP did not inhibit activity-dependent neurotrophic factor-induced growth. These data indicate that an activity-dependent neurotrophic factor-like substance is an endogenous embryonic growth factor and that VIP-regulated growth occurs, at least in part, through activity-dependent neurotrophic factor. PMID- 10676439 TI - Regulation of postimplantation mouse embryonic growth by maternal vasoactive intestinal peptide. AB - Vasoactive intestinal peptide (VIP) is an identified regulator of growth in the embryonic day (E) 9-11 mouse. Mouse embryonic and extra-embryonic tissues were studied to identify the source of VIP at this critical time. VIP and mRNA was detected in the decidua/trophoblast at E8 and declined until E10, after which it was not detectable. VIP mRNA was not apparent in the embryo until E11-E12. At E9, cells in decidua had VIP as well as lymphocyte marker (delta and CD3) immunoreactivity. VIP binding sites were dense in the decidua/trophoblast at E6, which gradually decreased until E10. VIP binding sites were detected in embryonic neuroepithelium by E9. The transient presence of VIP binding sites and mRNA in the decidua/trophoblast correlate with the identified period of VIP growth regulation, when VIP mRNA is absent in the embryo. Therefore, these findings suggest that maternal decidual lymphocytes are the source of VIP that regulate early postimplantation embryonic growth. PMID- 10676440 TI - VIP neuroprotection against excitotoxic lesions of the developing mouse brain. AB - Intracerebral administration of the excitotoxin ibotenate to new-born mice induced white-matter lesions mimicking the periventricular leukomalacia occurring in human premature babies. In this model, co-injection of vasoactive intestinal peptide (VIP) prevented white-matter lesions. VIP did not prevent the initial appearance of white-matter lesion, but promoted a secondary repair with axonal regrowth. Co-administration of ibotenate, VIP, and transduction inhibitors showed that protein kinase C (PKC) and mitogen-associated protein kinase (MAPK) pathways were critical for neuroprotection. The combination of in vitro and in vivo studies suggested the following model: VIP activates PKC in astrocytes, which release soluble factors; these released factors activate neuronal MAPK and PKC, which will permit axonal regrowth. Previous studies had shown that VIP-treated cultured astrocytes release growth factors including activity-dependent neurotrophic factor (ADNF) and that a 14-amino-acid peptide derived from ADNF protected the developing white matter against ibotenate. However, co-treatment with ibotenate, VIP, and anti-ADNF antibodies did not abolish VIP-induced protection, suggesting that ADNF does not mediate VIP protective properties in the present model. PMID- 10676441 TI - A novel signaling molecule for neuropeptide action: activity-dependent neuroprotective protein. AB - The complete coding sequence of a novel protein (828 amino acids, pI 5.99), a potential new mediator of vasoactive intestinal peptide (VIP) activity was recently revealed. The expression of this molecule, activity-dependent neuroprotective protein (ADNP), was augmented in the presence of VIP, in cerebral cortical astrocytes. The mRNA transcripts encoding ADNP were enriched in the mouse hippocampus and cerebellum. The protein deduced sequence contained the following: (1) a unique peptide, NAPVSIPQ, sharing structural and immunological homologies with the previously reported, activity-dependent neurotrophic factor (ADNF) and exhibiting neuroprotection in vitro and in vivo; (2) a glutaredoxin active site; and (3) a classical zinc binding domain. Comparative studies suggested that the peptide, NAPVSIPQ (NAP), was more efficacious than peptides derived from ADNF. ADNP, a potential mediator of VIP-associated neuronal survival, and the new peptide, a potential lead compound for drug design, are discussed below. PMID- 10676442 TI - Endomorphins: novel endogenous mu-opiate receptor agonists in regions of high mu opiate receptor density. AB - Endomorphin-1 (Tyr-Pro-Trp-Phe-NH2, EM-1) and endomorphin-2 (Tyr-Pro-Phe-Phe-NH2, EM-2) are peptides recently isolated from brain that show the highest affinity and selectivity for the mu (morphine) opiate receptor of all the known endogenous opioids. The endomorphins have potent analgesic and gastrointestinal effects. At the cellular level, they activate G-proteins (35S-GTP gamma-S binding) and inhibit calcium currents. Support for their role as endogenous ligands for the mu opiate receptor includes their localization by radioimmunoassay and immunocytochemistry in central nervous system regions of high mu receptor density. Intense EM-2 immunoreactivity is present in the terminal regions of primary afferent neurons in the dorsal horn of the spinal cord and in the medulla near high densities of mu receptors. Chemical (capsaicin) and surgical (rhizotomy) disruption of nociceptive primary afferent neurons depletes the immunoreactivity, implicating the primary afferents as the source of EM-2. Thus, EM-2 is well-positioned to serve as an endogenous modulator of pain in its earliest stages of perception. In contrast to EM-2, which is more prevalent in the spinal cord and lower brainstem, EM-1 is more widely and densely distributed throughout the brain than EM-2. The distribution is consistent with a role for the peptides in the modulation of diverse functions, including autonomic, neuroendocrine, and reward functions as well as modulation of responses to pain and stress. PMID- 10676443 TI - The ontogeny of endomorphin-1- and endomorphin-2-like immunoreactivity in rat brain and spinal cord. AB - Endomorphin-1 and endomorphin-2 are recently described peptides with high affinity and specificity for the mu opioid receptor. They are believed to be the endogenous ligands for that receptor. We describe the maturation of the endomorphins in brain and spinal cord using recently characterized antibodies to each. Endomorphin-1-like immunoreactivity was examined in brain, focusing on the periaqueductal gray of the midbrain and the diagonal band of Broca; endomorphin-2 like immunoreactivity is reported for the medulla and spinal cord. In these regions, and in all other regions studied but not described in this paper, the endomorphins were not seen at birth or at 3 days of age. Staining was present in 7-day-old and older animals. At these early ages, the pattern and density of staining are not fully developed, but appear complete by 21 days of age. The results suggest that both endomorphin-1 and endomorphin-2 develop relatively late compared to other opioid peptides. PMID- 10676444 TI - Pain inhibition by endomorphins. AB - Spinal analgesic effects of endomorphin-1 and endomorphin-2 were studied during acute, inflammatory, and neuropathic pain in rats chronically implanted with intrathecal cannulas. Endomorphin-1 and endomorphin-2 (2.5-10 micrograms i.t.), as well as their analogues, increased the tail-flick and the paw pressure latencies. In a model of inflammatory pain, the formalin-induced behavior was attenuated by endomorphins; however, the effect studied was not dose-dependent and was less pronounced in comparison with that evoked by morphine. On the other hand, in rats with a sciatic nerve injury (crush), endomorphins antagonized allodynia in a dose-dependent manner, whereas morphine was found to be ineffective in a similar dose range. Endomorphins also exhibited an antinociceptive potency in rats tolerant to morphine. In conclusion, our results show a powerful analgesic action of endomorphins at the spinal level. The most interesting finding is a strong effect of endomorphins in neuropathic pain, which opens up a possibility of using these compounds in pain therapy. PMID- 10676445 TI - Vasodilator responses to the endomorphin peptides, but not nociceptin/OFQ, are mediated by nitric oxide release. AB - The endomorphin peptides, endogenous ligands for the mu-opioid receptor, and nociceptin (orphanin FQ; OFQ), an endogenous ligand for the ORL1 receptor, have substantial vasodilator activity in the rat. The roles of nitric oxide, vasodilator prostaglandins, and the opening of K+ATP channels in mediating vasodilator responses to these novel agonists were investigated in the hindquarters vascular bed of the rat. Under constant-flow conditions, injections of the mu-selective agonists, endomorphin 1 and 2, PL017 ([N-MePhe3, D-Pro4] morphiceptin), and DAMGO, and the ORL1 receptor agonist, nociceptin/OFQ, produced dose-dependent decreases in hindquarters perfusion pressure. Vasodilator responses to endomorphin 1, PL017, and DAMGO were attenuated by the nitric oxide synthase inhibitor L-NAME at a time when vasodilator responses to nociceptin/OFQ were not altered. Responses to endomorphin 1 and 2, PL017, DAMGO, and nociceptin/OFQ were not altered by the cyclooxygenase inhibitor sodium meclofenamate or the K+ATP channel blocker U-37883A. The results of these studies indicate that responses to endomorphin 1 and 2, PL017, and DAMGO are mediated in large part by the release of nitric oxide, while responses to nociceptin/OFQ are mediated by an L-NAME-insensitive mechanism. Moreover, these results demonstrate that responses to these peptides are not mediated by the release of vasodilator prostaglandins or K+ATP channel opening in the hindquarters vascular bed. PMID- 10676446 TI - Brain tachykinins and the regulation of salt intake. AB - The regulation of salt intake is achieved through the coordination of behavioral and physiological responses. Brain neuropeptides, such as the tachykinins, play an important role in orchestrating both of these responses. Intraventricular injections of NK3 receptor agonists, such as senktide, are potent in suppressing salt intake. Experimental results show that intraventricular injections of senktide that suppress salt intake have no effect on the ingestion of other tastes, such as sucrose. The means by which senktide suppresses salt intake was investigated in a series of experiments. Taste reactivity and lick rate analyses suggest that the activation of NK3 receptors reduces salt intake by modulating the oral-stimulating property of salt taste. PMID- 10676447 TI - Neurotransmitter interaction in release of intranuclear oxytocin in magnocellular nuclei of the hypothalamus. AB - Oxytocin (OT) is released within the paraventricular (PVN) and supraoptic (SON) nuclei of the hypothalamus in response to several stimuli. However, the neurotransmitters that control this intranuclear OT release are unknown. In vivo microdialysis was used to examine the roles of norepinephrine and histamine in intranuclear OT release in conscious, lactating female rats. Administration of alpha- or beta-noradrenergic agonists, or histamine, increased OT release in the PVN. In addition, the increase in PVN OT evoked by exogenous histamine was prevented by simultaneous blockade of either H1 or H2 receptors. Furthermore, histamine-induced release of intranuclear OT was also prevented by blockade of alpha-adrenergic receptors. Finally, the increase in magnocellular OT release induced by suckling was abolished by administration of alpha-adrenergic antagonists. These data demonstrate that norepinephrine and histamine are important neurotransmitters for release of intranuclear OT, and histamine releases intranuclear OT by stimulating norepinephrine release. PMID- 10676448 TI - The role of brain-gut peptides in the control of sodium appetite. AB - Ingestion of food and fluid stimulates release of a number of peptides from the gastrointestinal system. These peptides are recognized to act as neurotransmitters/neuromodulators and act at both peripheral and central receptors. Many studies indicate that these peptides are important signals in terminating meals. Recent studies suggest that bombesin, a peptide related to gastrin-releasing peptide, suppresses sodium appetite. We have investigated the role of cholecystokinin (CCK) in the control of sodium appetite. Our studies indicate that CCK is effective at reducing saline intake. We found that exogenous, intraperitoneal CCK octapeptide suppresses saline intake. Moreover, administration of trypsin inhibitor to stimulate endogenous CCK release resulted in suppression of saline intake. Finally, intraperitoneal administration of the CCK receptor antagonist lorglumide resulted in increased saline intake. These observations extend the potential role of gastrointestinal peptides in the modulation of ingestive behavior. PMID- 10676449 TI - Endogenous vasopressin modulates the cardiovascular responses to exercise. AB - The role of brain-stem vasopressinergic projections in the genesis of reflex bradycardia and in the modulation of heart rate control during exercise is discussed on the basis of both changes in endogenous peptide content and heart rate changes observed during exercise. Dynamic running caused an increase in vasopressin content specifically in dorsal and ventral brain-stem areas. Rats pretreated with vasopressin or the V1 receptor antagonist into the nucleus tractus solitarii (NTS) showed a significant potentiation or a marked blunting of the exercise tachycardia, respectively, without any change in the blood pressure response. It is proposed that long-descending vasopressinergic pathways from the hypothalamus to the NTS serves as one link between the two main neural controllers of the circulation, that is, the central command and feedback control mechanisms driven by the peripheral receptors signals. Therefore vasopressinergic input contributes to the adjustment of heart rate response (and cardiac output) to the circulatory demand during exercise. PMID- 10676450 TI - Pharmacology of FMRFamide-related peptides in helminths. AB - Nervous systems of helminths are highly peptidergic. Species in the phylum Nematoda (roundworms) possess at least 50 FMRFamide-related peptides (FaRPs), with more yet to be identified. To date, few non-FaRP neuropeptides have been identified in these organisms, though evidence suggests that other families are present. FaRPergic systems have important functions in nematode neuromuscular control. In contrast, species in the phylum Platyhelminthes (flatworms) apparently utilize fewer FaRPs than do nematodes; those species examined possess one or two FaRPs. Other neuropeptides, such as neuropeptide F (NPF), play key roles in flatworm physiology. Although progress has been made in the characterization of FaRP pharmacology in helminths, much remains to be learned. Most studies on nematodes have been done with Ascaris suum because of its large size. However, thanks to the Caenorhabditis elegans genome project, we know most about the FaRP complement of this free-living animal. That essentially all C. elegans FaRPs are active on at least one A. suum neuromuscular system argues for conservation of ligand-receptor recognition features among the Nematoda. Structure-activity studies on nematode FaRPs have revealed that structure activity relationship (SAR) "rules" differ considerably among the FaRPs. Second messenger studies, along with experiments on ionic dependence and anatomical requirements for activity, reveal that FaRPs act through many different mechanisms. Platyhelminth FaRPs are myoexcitatory, and no evidence exists of multiple FaRP receptors in flatworms. Interestingly, there are examples of cross phylum activity, with some nematode FaRPs being active on flatworm muscle. The extent to which other invertebrate FaRPs show cross-phylum activity remains to be determined. How FaRPergic nerves contribute to the control of behavior in helminths, and are integrated with non-neuropeptidergic systems, also remains to be elucidated. PMID- 10676451 TI - Actions of nematode FMRFamide-related peptides on the pharyngeal muscle of the parasitic nematode, Ascaris suum. AB - The endogenous nematode peptides known as FMRFamide-related peptides (FaRPs) and various "classical" transmitters have a range of effects on nematodes that result in changes in behavior, particularly locomotion, including paralysis and inhibition of feeding. This study describes the application of an in vitro pharmacological approach to further delineate the action of a number of FaRP neurotransmitters on feeding behavior. Contraction of Ascaris suum pharyngeal muscle was monitored using a modified pressure transducer system that detects changes in intrapharyngeal pressure and therefore contraction of the radial muscle of the pharynx. The pharynx did not contract spontaneously. However, serotonin (5-HT, 100 microM) stimulated rhythmic contractions and relaxations (pumping) at a frequency of 0.5 Hz. The native nematode peptide, KNEFIRFamide (AF1), inhibited the pumping elicited by 5-HT. The duration of inhibition was concentration-dependent (1-1000 nM) with a threshold of 1 nM (n = 7). KSAYMRFamide (AF8/PF3) also inhibited pharyngeal pumping. There was no observable effect of any of the following nematode peptides on pharyngeal pumping behavior (1-1000 nM; n = 8): AF2, AF3, AF4, AF6, AF16, PF1/CF1, PF2/CF2, or PF4. Thus, interruption of pharyngeal processes, such as feeding, regulation of hydrostatic pressure, and secretion, may provide a new site of anthelmintic action. PMID- 10676452 TI - Neuropeptide gene families in the nematode Caenorhabditis elegans. AB - Neuropeptides have diverse roles in the function and development of the nervous system. With the completion of the sequencing of the C. elegans genome, rapid identification of nematode neuropeptide genes is possible. To date, 41 C. elegans neuropeptide genes have been identified. Of these genes, 20 genes, named flp (FMRFamide-like peptide) genes, encode FMRFamide-related proteins (FaRPs). Deletion of one of the flp genes, flp-1, results in several behavioral defects, suggesting that at least one flp gene is not functionally redundant with other flp genes. Twenty-one genes, named neuropeptide-like protein (nlp) genes, encode peptides distinct from the FaRP family. The predicted nlp-1 and nlp-2 neuropeptides have modest similarity to buccalin and myomodulin, respectively. Cellular expression patterns and genetic analysis of flp and nlp genes suggest that neuropeptides in nematodes also have widespread and varied roles in nervous system function. PMID- 10676453 TI - Comparison of FaRP immunoreactivity in free-living nematodes and in the plant parasitic nematode Heterodera glycines. AB - The family of FMRFamide-related peptides (FaRPs) is widely distributed among invertebrates, where the peptides serve as neuromodulators. Published reports indicate that numerous FaRP sequences exist in free-living and animal parasitic nematodes. Using a FMRFamide ELISA, FaRP immunoreactivity was detected in extracts of the soybean cyst nematode, Heterodera glycines, in both sexes and at all developmental stages. HPLC-ELISA results revealed a number of immunoreactive components in H. glycines preparations, and a comparison with extracts of the free-living nematodes Caenorhabditis elegans and Panagrellus redivivus showed significant qualitative differences in FaRP immunoreactivity between the plant parasite and the two free-living nematodes. Total and specific immunoreactivities varied during H. glycines development, with the highest specific activity in juveniles and males, and the highest total activity in mature females. Total female immunoreactivity was located primarily within the mature eggs. A significant portion, however, was associated with the female body, perhaps with egg laying. PMID- 10676454 TI - Structure, function, and expression of Drosophila melanogaster FMRFamide-related peptides. AB - In 1977, Price and Greenberg identified the tetrapeptide FMRFamide as a cardioexcitatory molecule from mollusc. Subsequent to this discovery, FMRFamide related peptides (FaRPs) have been identified in both invertebrates and vertebrates. Peptides in the FaRP family contain a common RFamide C-terminus and act as modulators and messengers of neural and gastrointestinal functions. Like other organisms, Drosophila melanogaster contains several genes that encode for numerous FaRPs. Elucidating the processing and activities of multiple FaRPs encoded in a single precursor is critical to establishing their roles in physiology. In this manuscript, we describe the distribution of FMRFamide immunoreactive materials in the Drosophila central nervous system and gut, and correlate it with the expression of specific FaRPs and their activities. The unique distributions and biological activities of Drosophila FaRPs suggest that the precursors are highly processed and the structurally related peptides are not functionally redundant. The complete distribution of FaRPs in the central nervous system and gut as detected by FMRFamide antisera is not accounted for by the sum of the individual expression patterns of the known Drosophila peptides. Thus, these data suggest that one or more Drosophila FaRPs or structurally related peptides remain to be discovered. PMID- 10676455 TI - Stimulation of alpha-amylase release in the scallop Pecten maximus by the myosuppressins. Structure-activity relationships. AB - The insect myosuppressin LMS (pGlu-Asp-Val-Asp-His-Val-Phe-Leu-Arg-Phe-NH2) elicits potent stimulation of the release of the digestive enzyme alpha-amylase from cell suspensions of the stomach-digestive gland complex of the scallop Pecten maximus. The myosuppressins are members of the FMRFamide-like peptide superfamily, which immunocytochemical data confirm is present in the scallop. Structure-activity studies indicated that the two most critical residues for bioactivity are Arg and Phe. Bioactivity of the peptide can be maintained if the basic, aromatic residue His is replaced by another basic residue (Lys) and another aromatic residue (Trp), but not the aromatic Tyr, indicating a sensitivity to the introduction of a phenolic OH group. A restricted-conformation analogue containing a cyclopropyl-Ala residue in position 8 (Cpa-MS) demonstrates an ability to antagonize the amylase secretion activity of LMS at microM concentrations. This result provides evidence that the myosuppressins adopt a tight turn in the C-terminal tetrapeptide active core region while binding to the scallop digestive gland receptor. Cpa-MS may provide a useful tool to neuroendocrinologists studying in vitro and in vivo digestive processes in mollusks and other invertebrates. PMID- 10676456 TI - The unique neuropeptide pattern in abdominal perisympathetic organs of insects. AB - We successfully isolated and identified the abundant neuropeptides of the abdominal perisympathetic organs of the American cockroach, including all myoactive compounds. Peptide sequence analysis and mass spectrometry of abundant substances that were not bioactive in different muscle assays yielded the following sequences: TDPLWQLPGAHLEQYLS-NH2 (Pea-YLS-amide), AFLTLTPGSHVDSYVEA-OH (Pea-VEAacid), and SDLTWTYQSPGDPTNSKN-OH (Pea-SKNacid). The given structures led to the conclusion of an unique neuropeptide pattern in abdominal perisympathetic organs. We confirmed this assumption with immunocytochemical studies, using antisera raised against different myotropic neuropeptides of the abdominal perisympathetic organs. Moreover, mass spectrometric methods, developed for the investigation of single neurohemal organs, confirmed the neuropeptide pattern in these organs. PMID- 10676457 TI - Adipokinetic hormones. Coupling between biosynthesis and release. AB - During long-distance flight of migratory locusts, the dramatic energy demand of the flight muscles is controlled by three adipokinetic hormones (AKHs). These peptide hormones regulate the mobilization of lipid and carbohydrate stored in the fat body to serve as energy substrates for the flight muscles. Despite the relatively huge quantities of the three AKHs that are stored in the corpora cardiaca, flight induces a differential 2-4-fold increase in the mRNAs for the three hormones. Moreover, newly synthesized AKHs can be released only during a restricted period of time, suggesting that by far most of the stored hormones are physiologically inactive. This raises the question of how the biosynthetic activity in the AKH-producing cells is coupled to their secretory activity. The present review discusses the potential mechanisms by which generation and release of mixtures of bioactive neurohormones are controlled and how peptidergic neuroendocrine cells cope with variations in physiological stimulation, with the AKH-producing cells serving as a model system. PMID- 10676458 TI - The regulation of juvenile hormone production in arthropods. Functional and evolutionary perspectives. AB - Although sesquiterpenoids are probably the ancestral regulators of reproduction and secondarily of metamorphosis in arthropods, our discussion suggests that the neuropeptides that regulate the biosynthesis of these compounds have arisen on several distinct occasions. These peptides probably occurred originally as regulators of other physiological processes and were subsequently co-opted for the regulation of sesquiterpenoid biosynthesis, perhaps first in adult forms and thereafter in larval forms. The evolution of peptides to assume additional physiological functions probably occurred as a result of gene duplication, both at the peptide level and at the receptor level. There are likely to be numerous regulators of sesquiterpenoid biosynthesis in both Insecta and Crustacea, and investigations to date have only begun to reveal the host of peptide families involved in the regulation of juvenile hormone-related biosynthesis across the arthropods. PMID- 10676459 TI - Allatostatins: a growing family of neuropeptides with structural and functional diversity. AB - The high degree of conservation of the core sequence of the "cockroach-types" of AST and their widespread distribution suggest that they should be considered a ubiquitous family of peptides within the invertebrates, regulating a range of important physiological processes. These functional processes, by either neural or humoral routes of action, include the inhibition of endocrine function, interneuronal functions, neuromodulatory roles, myotropic and myoendocrine roles, and direct action on biosynthetic pathways. The myomodulatory function appears to be conserved through evolutionary time, whereas the JH inhibitory activity appears to be confined to specific orders. This suggests that the myomodulatory role was the more ancestral of these two particular functions. Certainly, further purification and gene cloning as a means to precursor identification and functional analysis will be a prerequisite to understanding the diverse functions of this peptide family. PMID- 10676460 TI - The significance of Manduca sexta allatostatin in the tomato moth Lacanobia oleracea. AB - The nature and regulation of juvenile hormone (JH) biosynthesis have been investigated in isolated corpora allata (CA) of adult males and females, and larvae of the tomato moth Lacanobia oleracea (Noctuidae). Larval CA (Vth day 1 to VIth day 1) appear to synthesize JH at very low rates (< 0.5 fmol/pr/h). This synthesis was not affected by Manduca sexta allatostatin (Mas-AS) nor Manduca sexta allatotropin. In contrast, adult female CA synthesize relatively high levels of JHI and JHII (> 10 pmol/pr/h), each of which can be inhibited (approx. 60%) by Mas-AS. CA from adult male L. oleracea do not produce detectable levels of JH but would appear to synthesize JH-acids instead, which can also be inhibited (approx. 50%) by Mas-AS. When assayed on adult female L. oleracea CA, brain extracts separated by liquid chromatography show inhibitory activity. The major biologically active fraction also has the greatest Mas-AS-like immunoreactivity and co-elutes with synthetic Mas-AS, indicating that most of the allatostatic activity in brain extracts is due to a Mas-AS-like peptide. In adult male brains, even though relatively high levels of immunoreactivity co-elute with synthetic Mas-As, the majority of the Mas-AS-like immunoreactivity is more hydrophobic. PMID- 10676461 TI - Insect angiotensin-converting enzyme. A processing enzyme with broad substrate specificity and a role in reproduction. AB - Insect angiotensin-converting enzyme (ACE) is a peptidyl dipeptidase that removes dipeptides and dipeptideamides from the C-terminus of a broad range of in vitro oligopeptide substrates. In mammals, ACE has important roles in blood homeostasis and a recently recognized, but as yet undefined, role in the fertility of male mice. High levels of ACE are found in the male reproductive tissues of several insect species, and emerging data indicates an important role for the enzyme in insect reproduction. In this paper we review some of the recent findings about insect ACE, and we speculate as to the physiological role of this enzyme in insect reproduction. PMID- 10676462 TI - Development of amphiphylic mimics of insect neuropeptides for pest control. AB - Insect neuropeptides regulate virtually all aspects of insect life and are excellent candidates for development of new methods for pest control. However, they do not penetrate insect cuticle and are degraded by enzymes both in the digestive system and hemolymph. We have designed mimics of model neuropeptides by attachment of various lipidic moieties to the amino terminus of the bioactive core of the neuropeptides. These mimics have amphiphylic characteristics that allowed them to penetrate the hydrophobic insect cuticle. The mimics also induced prolonged physiological responses (up to 20 hours) and were resistant to peptidase attack. This knowledge has been used to develop a novel, species specific approach to insect control. PMID- 10676463 TI - The kinin peptide family in invertebrates. AB - Kinins comprise a family of peptides that were first found in the central nervous system of insects and recently also in mollusks and crustaceans. After the isolation of the first members of the kinin family, the leukokinins from Leucophaea maderae, leukokinin-related peptides were found in the cricket Acheta domesticus and the locust Locusta migratoria, all through their ability to induce Leucophaea maderae hindgut contraction. Subsequently, kinins were found in the mosquitoes Culex salinarius and Aedes aegypti and in the earworm Helicoverpa zea. The first noninsect member of this family was isolated from a mollusk, the pond snail Lymnaea stagnalis. Most recently our group has isolated the first kinins from crustaceans. Six kinins were isolated from the white shrimp Penaeus vannamei. To date, 35 members of this family have been isolated. The first relatively small family of insect kinins has grown into an expanding and rather large family with members in insects, crustaceans, and mollusks. In this paper we discuss the kinin family in terms of method of isolation, structure, in vitro and in vivo activity, distribution, receptors, and signal transduction. We will compare the crustacean and insect members of the kinin family, using the data available on crustacea. PMID- 10676464 TI - Tachykinin-like peptides and their receptors. A review. AB - Tachykinin-like peptides have been identified in many vertebrate and invertebrate species. On the basis of the data reviewed in this paper, these peptides can be classified into two distinct subfamilies, which are recognized by their respective sequence characteristics. All known vertebrate tachykinins and a few invertebrate ones share a common C-terminal sequence motif, -FXGLMa. The insect tachykinins, which have a common -GFX1GX2Ra C-terminus, display about 30% of sequence homology with the first group. Tachykinins are multifunctional brain/gut peptides. In mammals and insects, various isoforms play an important neuromodulatory role in the central nervous system. They are involved in the processing of sensory information and in the control of motor activities. In addition, members of both subfamilies elicit stimulatory responses on a variety of visceral muscles. The receptors for mammalian and insect tachykinins show a high degree of sequence conservation and their functional characteristics are very similar. In both mammals and insects, angiotensin-converting enzyme (ACE) plays a prominent role in tachykinin peptide metabolism. PMID- 10676465 TI - Comparison of active conformations of the insectatachykinin/tachykinin and insect kinin/Tyr-W-MIF-1 neuropeptide family pairs. AB - A comparison of solution conformations of active, restricted-conformation analogues of two sequence-similar insect/vertebrate neuropeptide family pairs shed light on the potential existence of molecular evolutionary relationships. Analogues of the locustatachykinins and the mammalian tachykinin substance P, containing a sterically hindered Aib-NMePhe/Tyr residue block, share similar low energy turn conformations incorporating a cis peptide bond. Conversely, restricted conformation analogues of the insect kinins and the mammalian opiate peptide Tyr-W-MIF-1, with near identical C-terminal tetrapeptide sequences, adopt different conformations. The insect kinins adopt a cisPro 1-4 beta-turn, in which the Phe1 is critical for bioactivity. Tyr-W-MIF-1 prefers a transPro 2-5 turn, and an additional N-terminal Phe severely inhibits mu-opiate receptor binding. Comparisons of the chemical/conformational requirements for receptor interaction are consistent with a distant evolutionary relationship between the insectatachykinins and tachykinins, but not between the insect kinins and Tyr-W MIF-1. Therefore, analogues of the insect kinins with pest control potential can be readily designed to avoid mammalian interactions. PMID- 10676466 TI - VIP and PACAP38 modulate cytokine and nitric oxide production in peritoneal macrophages and macrophage cell lines. PMID- 10676467 TI - Axonal plasticity in the rat pituitary intermediate lobe following chronic D2 receptor modulation. PMID- 10676468 TI - Chronic intracerebroventricular infusion of beta-amyloid (1-40) results in a selective loss of neuropeptides in addition to a reduction in choline acetyltransferase activity in the cortical mantle and hippocampus in the rat. PMID- 10676469 TI - Correlates of deoxycorticosterone-induced salt appetite behavior and basal ganglia neurochemistry. PMID- 10676470 TI - Co-localization of the inwardly rectifying potassium ion channel, Kir2.2, and the substance P receptor in single locus coeruleus neurons. PMID- 10676471 TI - Intraventricular injections of tachykinin NK3 receptor agonists affect salt intake. A shift in taste intensity? PMID- 10676472 TI - Dominance of nonphotic cues in the circadian rhythm of body temperature in vasopressin-deficient rats. PMID- 10676473 TI - Acquisition of improved reference values for cesium, iodine, strontium, thorium, and uranium in selected NIST reference materials. AB - As part of a study on the ingestion and organ content of some trace elements of importance in radiological protection, additional work has been undertaken to acquire improved reference values for cesium, iodine, strontium, thorium, and uranium in four selected reference materials provided by the US National Institute of Standards and Technology. The materials are SRM-1548 Total Diet, SRM 1548a Typical Diet, SRM-1486 Bone Meal, and RM-8414 Bovine Muscle. A coordinated study was undertaken with the help of seven selected laboratories in five countries. Instrumental and radiochemical neutron activation analysis and inductively coupled plasma-mass spectrometry were the analytical main techniques used. PMID- 10676474 TI - Trends in trace element determinations in blood, serum, and urine of the Dutch population, and the role of neutron activation analysis. AB - A critical review on the quality of literature data on trace elements in human blood, serum, and urine of inhabitants in the Netherlands has shown that many of the currently available data have been established 15-20 years ago. Only in a few publications are quality indicators mentioned, which should be considered typical -and minimal--for studies resulting at reference values. The use of neutron activation analysis for determination of trace elements in human body fluids was restricted to a few studies in the 1970s. However, although it is frequently assumed that the sensitivity of NAA might be insufficient, it is demonstrated that modern, large, well-type Ge detectors may serve well for the determination of trace elements in human body fluids via radiochemical NAA, for example. PMID- 10676475 TI - INAA of serum zinc of inhabitants in five regions of the Czech Republic. AB - To investigate the Zn status of inhabitants of the Czech Republic, 1155 serum and 132 hair samples were analyzed for zinc content. Analyzed material was obtained from randomly selected volunteers of both sexes in the age range 6-65 yr. Subpopulations from five regions were included in the study. Analyses of both materials were performed by Instrumental Neutron Activation Analysis (INAA). Coanalyses of Standard Reference Materials (SRM) for quality control were performed. The results (mean 910 +/- 276 micrograms Zn/L serum and 189 +/- 45 micrograms Zn/g hair) demonstrate satisfactory zinc status of the searched population. Significant interregional differences and age-dependent differences, as well as sex-dependent differences have been detected by the use of correlation analysis. On the basis of these results, serum Zn concentration results of individual categories according to age and sex were evaluated, and on the grounds of mean +/- 2 standard deviations, indicative intervals were calculated. The frequency of individual serum Zn concentrations proved that the population of the Czech Republic does not suffer from severe zinc deficiency. On the other hand, about one-third of our inhabitants has their serum Zn concentrations below the cutoff value of 800 micrograms Zn/L serum, which means a marginal or mild Zn deficit of the organism. PMID- 10676476 TI - Characterization of selenium status of inhabitants in the region Usti nad Orlici, Czech Republic by INAA of blood serum and hair and fluorimetric analysis of urine. AB - Selenium is an essential trace element and its insufficient status may cause serious health complications for both individuals and the whole populations. To investigate the selenium status of the subpopulation in northeastern Bohemia represented by the region Usti nad Orlici, 253 serum, 469 urine, and 31 hair samples from 470 randomly selected volunteers between 6 and 65 yr of age have been analyzed for selenium concentration. Serum and hair Se were detected by instrumental neutron activation analysis (means: 55 +/- 11 micrograms Se/L sera, 0.268 +/- 0.040 microgram Se/g hair). Urine Se was measured by fluorimetry (12 +/ 5 micrograms Se/L urine) with coanalyses of Lyphocheck urine, SRM Urine 2670, and Seronorm urine for quality control of the method. Results proved significant age-dependent differences, but gender differences were not significant. The frequency plot of serum Se proved maximal frequencies in adults between 55 and 70 micrograms Se/L and in children in the range 45-55 micrograms Se/L. The same plots of urine Se for both age groups showed maximal frequency in the limits 8-15 micrograms Se/L. All indices used (Se in serum, urine, and hair) confirmed mild to severe selenium deficiency in the population of the region. PMID- 10676477 TI - Instrumental neutron activation analysis of rib bone samples and of bone reference materials. AB - The instrumental neutron activation analysis method was used for the determination of trace elements in rib bone samples taken from autopsies of accident victims. The elements Br, Ca, Cl, Cr, Fe, Mg, Mn, Na, P, Sr, Rb, and Zn were determined in cortical tissues by using short and long irradiations with thermal neutron flux of the IEA-R1m nuclear reactor. The reference materials NIST SRM 1400 Bone Ash and NIST SRM 1486 Bone Meal were also analyzed in order to evaluate the precision and the accuracy of the results. It was verified that lyophilization is the most convenient process for drying bone samples because it does not cause any element losses. Comparisons were made between the results obtained for rib samples and the literature values as well as between the results obtained for different ribs from a single individual and for bones from different individuals. PMID- 10676478 TI - IAEA Internet database of natural matrix reference materials. AB - The International Atomic Energy Agency maintains a database on internationally available certified reference materials of natural origin. The database was updated in 1998 and prepared for an Internet implementation. A user-friendly structure was created, providing two main pathways for browsing, either according to the matrix classification or the producer's name. The database presently contains over 20,000 values for 480 measurands and 1085 reference materials from 43 different producers. Most of the materials entered contain values for trace and minor elements (66%). PMID- 10676479 TI - Microbeam PIXE analysis of Na, K, Mg, and Ca in severely damaged cardiac tissue. AB - The aim of the study was to measure with microbeam PIXE elements such as Na, K, Mg, and Ca in cardiac tissue after various treatments in vivo, which affect the cardiomyocyte integrity. It was assumed that local deviations from normal electrolyte levels indicate the degree of cardiac damage. The first step in this feasibility study was comparison of severely damaged cardiac tissues with controls. Severe cardiac damage was introduced by the so-called Ca paradox. Experiments were performed with isolated rat hearts, perfused retrogradely with an oxygenated crystalloid buffer. Results indicated that severe cardiac damage was accompanied with almost complete disappearance of the normal intracellular electrolyte composition as a result of the loss of membrane integrity. Identifications of smaller and more locally present ischemic damages on basis of altered electrolyte levels appeared to be feasible. However, the prerequisite was that the mobility of electrolytes be kept under control during tissue sampling and sample preparation, when physiological mechanisms stop to maintain gradients. PMID- 10676480 TI - Nuclear microscopy in the life sciences at the National University of Singapore. A review. AB - The nuclear microscope is now gaining popularity in the field of life sciences. In particular, the combination of proton-induced X-ray emission to measure the elemental concentrations of inorganic elements, Rutherford backscattering spectrometry to characterize the organic matrix, and scanning transmission ion microscopy to provide information on the density and structure of the sample represents a powerful set of techniques that can be applied simultaneously to the specimen under investigation. These techniques are extremely useful for measuring any imbalances in trace elements in localized regions of biological tissue and, as such, can provide unique information on many diseases. In this article, we describe the nuclear microscope and its related ion-beam techniques, and we review the biomedical work carried out using the nuclear microscope in the National University of Singapore. PMID- 10676481 TI - An external ion microbeam for studies of biological samples. AB - A new external ion-beam system was developed and combined with a light ion microbeam system in JAERI Takasaki. The system is designed for micro-particle induced X-ray emission analysis of biological samples in air environment with 1 micron spatial resolution. One of the most serious problems in keeping such a high spatial resolution is multiple scattering in a beam exit window. Thin Kapton film (7.5 microns thick) was adopted as the exit window as well as a sample backing foil to minimize the distance between the film and samples. The lifetime of the foil under ion irradiation and spatial resolution of the external microbeam were investigated. The results shows that the film can endure sufficient long-time irradiation to take elemental maps and the resolution can be kept nearly 1 micron. PMID- 10676482 TI - Improvements in the determination of radionuclide distribution in the analysis of irradiated samples using double-differentiation method. AB - The spatial response of an 8 x 4 block detector made up of 5.6-mm-wide, 12.9-mm high, 30-mm-thick individual detector crystals to a collimated line source of 511 keV annihilation photons was examined. The response of each crystal showed a spread around the average positioning values and distributions from adjacent crystals overlapped as the collimated source scanned the individual detectors. This leads to possible errors in the event assignment. The implementation of double differentiation or the second derivative method was proposed for the removal of scattered photons so as to reduce the overlap and, hence, avoid mis positioning. This method is a mathematical solution implemented when analysing the results. A curve in a spatial spectrum could be considered to be a function f(x), where x is the position. When double differentiation of f(x) is carried out, then the normalized curve d2f(x) appears with some reduction in the wings. It was shown that a reduction of the scattering contribution in the tails without overestimating the contribution of scattered events could be achieved by implementing a double-differentiation process. PMID- 10676483 TI - Radiotracer analysis of cadmium speciation in soil solutions using electrophoresis, dialysis, centrifugation, and ultrafiltration. AB - Speciation of carrier-free 109Cd, added in cationic form to pre-filtered extracts obtained by leaching forest soil samples with distilled water, was analyzed using electrophoresis, dialysis, centrifugation, and ultrafiltration. Rapid establishment of isotopic equilibrium between the added 109Cd and stable cadmium present in the extracts was observed. All the data obtained indicated that 109Cd and also stable cadmium were present in the analyzed extracts in the form of neutral or negatively charged organic complexes or small colloids. The results of electrophoresis enabled the characterization, at least semiquantitative, of the abundance and electrophoretic mobility of the forms present. The incomplete dialysis of 109Cd from the soil extracts through cellophane membrane against water proved the presence of organic associates with molecular weights higher than 10(4). Dialysis against the same, but unlabeled extract was always complete, indicating the reversible (labile) nature of the organic forms of cadmium. Assessment of the stability constants of the organic forms using a simple discrete two-site model suggested that humate and/or fulvate complexes of cadmium were formed. PMID- 10676484 TI - Study on binding of REEs with water-soluble polysaccharides in fern. AB - The binding of rare earth elements (REEs) with water-soluble polysaccharides of nondeproteinization and deproteinization in the leaves of the fern Dicranopteris dichotoma was studied by molecular activation analysis (MAA). Two cold-water soluble polysaccharides (extracted by 75% ethanol and 90% ethanol, respectively) and one hot-water-soluble polysaccharide (extracted by 90% ethanol) were separated using biochemical separation techniques. The eight rare earth elements (La, Ce, Nd, Sm, Eu, Tb, Yb, and Lu) in these polysaccharides were determined by instrumental neutron activation analysis. Our new results showed that the REEs were bound firmly with the water-soluble polysaccharides in the plant, regardless of whether nondeproteinization or deproteinization was used. The molecular-weight (MW) measurement demonstrated that REEs were mainly bound with low-MW (10,000 20,000) polysaccharides. PMID- 10676485 TI - Correlation between total and EDTA/DTPA-extractable trace elements in soil and wheat. AB - Wheat and wheat products are more important sources of energy and nutrients in diets of people in many cultures compared to other foods. The daily consumption of wheat is about 200 g/d/person in Western Europe and North America. On the other hand, 400-450 g of wheat and wheat products are consumed daily by average Turkish people. Wheat samples collected from the Iskenderun region in 1995 and 1996 and Ankara and Istanbul regions in 1995 were analyzed for their trace element content by instrumental neutron activation analysis (INAA). In addition, 13 soil samples were collected from the Iskenderun region in 1996. Total soil samples were analyzed by INAA and atomic absorption spectrometry (AAS), EDTA extractable elements by INAA, and DTPA-extractable elements by AAS. Correlation analysis and enrichment factor calculations were applied to the trace element results. In wheat samples, a strong correlation was found between the elements such as Sc, La, Sm, Rb, and K whose main source is soil. The concentration of Se appeared to show larger variations among different regions. No significant correlation was observed for elements such as As and Se whose main sources in the atmosphere are anthropogenic activities. PMID- 10676486 TI - Comparison of the chromium distribution in organs and subcellular fractions of normal and diabetic rats by using enriched stable isotope Cr-50 tracer technique. AB - The enriched stable isotope 50Cr(III) tracer technique combined with neutron activation analysis was used to examine the intracellular distribution of Cr(III) in the liver, pancreas, testes, and kidney homogenates of both normal and diabetic rats. Our new results showed that the nucleic fraction has the highest Cr concentration in the liver cell of both normal and diabetic rats. The diabetic rats retain more Cr in the mitochondrial and lysosomal fractions of liver homogenate than the normal. This is likely an indication of chromium participating in the glucose or lipid metabolism to compensate the low level of insulin in the body of diabetic rats. The concentrations of Cr in the subcellular fractions of pancreas, testes, and kidney in the normal rats are higher than those in the diabetic rats, which favor the hypothesis that Cr(III) plays its biological function via interaction with the insulin-sensitive tissues or enhancement of the sensitivity of the insulin receptor. PMID- 10676487 TI - Investigation of selenium distribution in subcellular fractions of human liver by neutron activation analysis. AB - Selenium is an important and essential trace element to living systems. In the article, two methods of instrumental neutron activation analysis and hydride generation-atomic fluorescence spectrometry were applied to determine Se in biological samples and the accuracy was evaluated by several reference materials. The subcellular distribution of selenium in human liver samples, which were obtained from normal subjects who had an accidental death, was investigated by differential centrifugation combined with INAA. Selenium was mainly enriched in mitochondria, nuclei, and cytosol. Almost half of the total Se content existed in nuclei as a result of the large amount in liver and the high Se concentration. Generally, the highest Se concentration in the mitochondrial fractions of each liver sample suggested that Se had important functions in this liver component. PMID- 10676488 TI - Speciation of inorganic arsenic and selenium in leachates from landfills in relation to water quality assessment. AB - Speciation of arsenic and selenium was carried out on water samples taken from rivers used as water intake points in the vicinity of landfill areas used for land-based waste disposal system. Leachates from these landfill areas may contaminate the river water through underground seepage or overflowing, especially after a heavy downpour. Preconcentration of the chemical species was done using a mixture of ammonium pyrrolidinethiocarbamate-chloroform (APDTC CHCl3). Because only the reduced forms of both arsenic and selenium species could be extracted by the preconcentrating mixture, suitable reducing agents such as 25% sodium thiosulfate for As(III) and 6M HCl for Se(IV) were used throughout the studies. Care was taken to exclude the interfering elements such as the alkali and alkali earth metals from the inorganic arsenic and selenium species by introducing 12% EDTA solution as the masking agent. The extracted mixture was irradiated in a thermal neutron flux of 4 x 10(12)/cm/s from a TRIGA Mk.II reactor at the Malaysia Institute of Nuclear Technology Research (MINT). Gamma rays of 559 keV and 297 keV from 76As and 75Se, respectively, were used in the quantitative determination of the inorganic species. Mixed standards of As(III) and Se(IV) used in the percentage efficiency procedure were prepared from salts of Analar grade. The water quality evaluation was viewed from the ratio of the inorganic species present. PMID- 10676489 TI - Spatial distribution of extractable organohalogens in northern pink shrimp in the north Atlantic. AB - Extractable organohalogens (EOX) are organic compounds that contain chlorine, bromine and/or iodine, which can be separated from the matrix by liquid/liquid or liquid/solid extraction. A combination of instrumental neutron activation analysis (INAA) and solvent extraction methods has been developed for the determination of EOX from the shrimp Pandalus borealis. Levels of EOX were evaluated for spatial trends for shrimp caught in several areas off the Labrador coast, off the coast of Nova Scotia, and off the coast of Maine. Muscle contained 1.09-6.05 micrograms EOCl/g tissue and 105-498 micrograms extractable organochlorine (EOCl)/g lipid; 0.0607-0.288 microgram extractable organobromine (EOB)r/g tissue and 4.74-10.5 micrograms EOBr/g lipid; and 0.014-0.048 microgram extractable organoiodine (EOI)/g tissue and 1.03-1.76 micrograms EOI/g lipid, respectively. The levels of EOCl in roe were 1.60-12.34 micrograms/g tissue and 39.0-146 micrograms/g lipid. In roe, the EOBr levels were 0.707-1.03 micrograms/g tissue and 6.96-13.5 micrograms/g lipid; and EOI levels were 0.123-0.349 microgram/g tissue and 1.42-4.11 micrograms/g lipid. The EOCl, EOBr, and EOI levels in roe increased noticeably from north to south along the coast of Labrador. Samples taken from the coast of Maine and from Canso Hole were typically higher in EOCl levels than those taken from Labrador. The results for EOBr and EOI were in the same range as those from Labrador. PMID- 10676490 TI - Health-related monitoring and assessment of airborne particulate matter. An overview of recent IAEA programs. AB - Since 1992, the International Atomic Energy Agency (IAEA) has been promoting studies of air pollution using a standard design of air sampler that provides separation on filters into two size fractions with cutoffs of 2.5 and 10 microns (approximately). These are the size ranges presently considered to have the most important health consequences. Such filter samples are highly amenable to analysis using nuclear and related techniques. After reviewing some of the health effects of airborne particulate matter and current air quality standards and guidelines, this article provides an overview of current and recent IAEA programs in this area, which involve collaborative activities with participants in more than 40 countries. PMID- 10676491 TI - Characterization of airborne particulates in Bangkok urban area by neutron activation analysis. AB - Samples of airborne particulates were collected in a residential area and in an area near a busy highway in Bangkok during the period from January 1997 to May 1998. A stacked filter system was used for the former site and a Partisol 2000 was used for the latter site. Both 2.5 microns and 10-micron particulates were collected every week. The total suspended particulate matters were also collected at the latter site. The samples were analyzed by neutron activation analysis utilizing neutron flux from a 2-MW TRIGA MARK III research reactor. The elements most frequently detected in the airborne particulates were Al, As, Br, Ca, Ce, Cl, Co, Cr, Cs, Fe, I, K, La, Mg, Mn, Na, Rb, Sb, Sc, Sm, Th, Ti, V, and Zn. The enrichment factor and factor analysis were used to investigate trends, sources, and origin of the atmospheric aerosols. Anthropogenic elements in road dust, construction dust, motor vehicles emission, and other combustion components were identified. A comparative study of data between both sites was performed and it was found that the mass concentration in the area close to the highway was about three times higher than in the residential area. PMID- 10676492 TI - A study of air particulate pollution in Colombo using a nuclear-related analytical technique. AB - Air particulate matter of two size fractions (coarse [2.5-10 microns] and fine [less than 2.5 microns]) were collected at an urban residential site (Colombo University ground) over a period of 12 mo during 1996 using a Gent PM10 stacked filter unit. Seventy-five sets of samples collected during this period were analyzed for 10 elements: Al, Si, K, S, Ca, Ti, Fe, Zn, Br, and Pb by energy dispersive X-ray fluorescence, which is a multielement analysis. This is a highly sensitive technique enabling quantitative analysis of very low masses. The average concentrations of lead, sulfur, and bromine, which are generally associated with combustion products of automotive exhausts, dominate the fine fraction in this study. The minimum and maximum concentration of lead resulting in an annual average of 0.09 microgram/m3 was found to be 0.0042 and 0.441 microgram/m3 in particulate matter with less than 10 microns equivalent aerodynamic diameter. The maximum concentration determined is well below the limits set by the World Health Organization (0.5-1.0 microgram/m3). The recommended value for Pb in Sri Lanka is 0.5 microgram/m3. Al, Si, Ca, Ti, and Fe exhibited enhanced concentrations in the coarse fraction, which probably originate from disturbed soil as a result of wind and traffic. PMID- 10676493 TI - Collection and characterization of a bulk PM2.5 air particulate matter material for use in reference materials. AB - A contemporary PM2.5 (particulate matter smaller than 2.5 microns aerodynamic diameter) aerosol material from an urban site has been collected for the production of a new standard reference material that will be made available for the development of new PM2.5 air quality standards. Air particulate matter corresponding to the PM2.5 fraction was collected at an established Environmental Protection Agency monitoring site in Baltimore, Maryland. The air-sampling system that has been constructed for this collection separates fine particles with a cyclone separator and deposits them onto an array of Teflon membrane filters. The fine air particulate material is removed by ultrasonication or by mechanical means and collected for further preparation of standards. The composition of the collected PM2.5 aerosol, as well as the composition of the deposited PM2.5 aerosol, are determined by instrumental nuclear activation analysis and other techniques. PMID- 10676494 TI - Determination of concentrations of elements in the atmospheric aerosol of the urban and rural areas of Beijing in winter. AB - The proton-induced X-ray emission technique is one of the most suitable methods in the study of the multielement content of atmospheric aerosols. Atmospheric aerosol samples were collected in winter using an eight-stage cascade impactor at a site of the urban center and a rural site of Beijing. The aerosol samples collected were analyzed to determine maximum, minimum, and average concentrations of up to 20 elements and the ratios of the average element concentrations for the coarse to fine particles (C/F) by the PIXE technique. It has been found that the average elemental concentrations in the urban center are higher than those in the rural area, except S and Br. The concentrations for S and Pb in the atmospheric aerosols are found to be less than the results of previous measurement, but their concentrations in the fine particles increased in winter in the samples from the urban center. The deposition probability of the International Commission on Radiological Protection (ICRP) mode and the mass particle-size distributions of the elements determined in the urban center have been utilized to evaluate inhalable particulate matter fractions retained in the three regions of one's respiratory tract and their harm to human health is discussed. PMID- 10676495 TI - Content of trace elements in the respirable fractions of the air particulate of urban and rural areas monitored by neutron activation analysis. AB - The concentrations (ng/m3) of more than 30 trace elements have been determined in the total air particulate matter and in the size-segregated fractions collected in urban, industrialized, and rural residential areas in northern Italy by means of a multistage inertial impactor with the PM10 inlet. All measurements have been carried out by instrumental neutron activation analysis, except for Pb and Cd, which have been determined by electrothermal atomic absorption spectroscopy. Analytical quality assurance procedures have been developed with special regard to blanks, reagents, and sampling. Total concentrations and the granulometric distribution found in the different locations are reported and compared. PMID- 10676496 TI - Measurement of PM10/2.5 fractionated respirable particles in urban Toronto by INAA, PIXE, ICP-AES, and LAMS. A comparison. AB - The chemical analysis of urban Toronto airborne particulate matter (PM), size segregated into respirable PM10/2.5, is presented. The characterization of PM by use of proton-induced X-ray emission analysis (PIXE), and inductively coupled plasma-atomic emission spectrometry revealed elemental information; a newer laser ablation-ionization mass spectrometry approach has the potential to expand the chemical information from PM by analyzing both the inorganic and organic species. These PM analytical approaches will be continued in the future for studying (1) emission source identification, (2) inhalation health hazards, and (3) urban smog chemistry. PMID- 10676497 TI - INAA and PIXE of atmospheric and combustion aerosols. AB - Using instrumental neutron activation analyses and photon-induced x-ray emission techniques for analysis of size-fractionated atmospheric and combustion aerosols and other emission samples arising from fluidized-bed combustion of North Bohemian lignites up to 42 elements were determined in all samples types. This allowed the evaluation of element enrichment, time trends, and inter-element correlations and the performance of factor analysis of various fractions of atmospheric aerosols. The data obtained on mass and element size distributions of aerosols and emission samples obtained upon lignite combustion in an experimental scale atmospheric fluidized-bed combustor without and with added hydrated lime and limestone were used to elucidate the mechanism of abatement of toxic trace and matrix elements from flue gas. PMID- 10676498 TI - An evaluation of atmospheric deposition of trace elements into the Great Lakes. AB - High-volume air samplers were used to collect aerosol samples on Whatman 41 air filters at the Canadian air sampling stations Burnt Island, Egbert, and Point Petre. Once collected, the samples were analyzed for trace elements by neutron activation analysis. Air concentrations of over 30 trace elements were determined. A special focus was made to utilize Compton suppression gamma-ray spectroscopy and epithermal irradiations to enhance the detection limits of neutron activation analysis. These techniques allowed for the determination of trace elements at very low levels. Results of the study of the trace-metal dry deposition into Lakes Huron and Ontario indicated that the majority of the total deposition resulted from crustal materials. However, dry deposition is also a significant pathway for many toxic anthropogenic trace metals into the Great Lakes. PMID- 10676499 TI - Biomonitoring of air pollution as exemplified by recent IAEA programs. AB - Biomonitoring is an appropriate tool for assessing the levels of atmospheric pollution, having several advantages compared with the use of direct measurements of contaminants (e.g., in airborne particulate matter, atmospheric deposition, precipitation), related primarily to the permanent and common occurrence of the chosen organisms in the field, the ease of sampling, and trace element accumulation. Furthermore, biomonitors may provide a measure of integrated exposure over an extended period of time and are present in remote areas and no expensive technical equipment is involved in collecting them. They accumulate contaminants over the exposure time and concentrate them, thus facilitating analytical measurements. Based on large-scale biomonitoring surveys, polluted areas can be identified, and by applying appropriate statistical tools, information can be obtained on the type of pollution sources and on the transboundary transport of atmospheric pollutants. The International Atomic Energy Agency is including the research on biomonitors in its projects on health related environmental studies. Biomonitoring activities from several coordinated research projects on air pollution are presented, and results from an international workshop are discussed. In addition, activities in supporting improvement quality in the participating laboratories are outlined. PMID- 10676500 TI - Study on air pollution in Beijing's major industrial areas using multielements in biomonitors and NAA techniques. AB - Three kinds of plant leaves, Chinese white aspen, arborvitae, and pine needles, have been sampled from the Yanshan Oil refinery complex, the Capital Iron and Steel Factory, and Badachu, a control area in Beijing, as biomonitors for air pollution studies. Each sample was divided into two parts, washed, and unwashed. Thirty-one trace elements (As, Au, Br, Ca, Cd, K, La, Lu, Mo, Na, Sb, Sm, U, W, Yb, Ba, Ce, Co, Cr, Cs, Eu, Fe, Hf, Hg, Rb, Sc, Se, Sr, Ta, Tb, Th, and Zn) have been determined by using the relative and the K0 methods of instrumental neutron activation analysis (NAA) techniques. The results indicated the following: (1) The concentration of trace elements in unwashed samples are much higher than these in washed samples; (2) the area around Capital Iron and Steel Factory is heavily polluted, and the Yanshan Oil refinery complex area is moderately polluted; (3) Chinese white aspen is a good biomonitor in particular seasons and pine needles are better than arborvitae for yearly monitoring; (4) elements As, Cd, Hg, Co, Rb, Sb, W, and Zn are highly absorbed by Chinese white aspen. Pine needles are sensitive for the absorption the elements Br, Cr, Cd, Fe, Sc, Cs and rare earth elements, but arborvitae is very sensitive for the absorption of Sr. PMID- 10676501 TI - Environmental assessment in an industrial area of Portugal. AB - The region of Lisbon and south of Lisbon (Sado estuary) is densely industrialized, and, therefore, air pollution should be studied in a more detailed scale there. The topography of the Sado estuary region and the predominant wind direction from the northwest contribute to the influence in this region of the industries located in the north. The region selected includes a fuel-fired power station. Transplants of the lichen Parmelia sulcata Taylor were suspended in nylon bags within a rectangle 15 km wide and 25 km long on a grid 2.5 km x 2.5 km, centered in the power station. In each of the 47 sites, 2 sets of 4 transplants each were hung. Care was taken (1) in covering the two sets with a polyethylene roof to prevent leaching of elements in the lichen, (2) in building a hanging system that could rotate according to the wind direction, and (3) in orienting one set toward the wind and the other set opposite the wind. For a 1-yr period and every 3 mo, one transplant of each set is collected. In this work, the results of the first campaign (after 3 mo suspension) obtained by instrumental neutron activation analysis and proton-induced X-ray emission are shown. Some elemental contents are mapped and discussed. PMID- 10676502 TI - Determinations of subnanomole elemental levels by NAA and their possible impact on human health related issues. AB - Neutron activation analysis (NAA) is an appropriate tool for the determination of trace elements in biological systems. The virtually blank-free NAA procedures fittingly complement precautions employed in sampling and sample preparation of biological matrices. Results from instrumental NAA procedures used to establish baseline values and time trends for elements in human tissues demonstrate the advantages as well as the limits of these procedures for nanomole and, in a considerable number of instances, subnanomole elemental levels. In addition, subnanomole mass fractions have been determined with extremely low limits of detection by employing NAA with radiochemical separations isolating very low levels of radioactivity from the matrix background. The elements reviewed in this article include Cr, Se, Pt, and others that have been determined by NAA at subnanomole levels in human tissues and body fluids and in biological macromolecules. PMID- 10676503 TI - Evaluation of trace elements in lung samples from coal miners using neutron activation analysis. AB - In this study, instrumental neutron activation analysis was applied to the determination of Sc, La, Ce, Nd, Sm, Eu, Tb, Yb, Lu, Hf, Th, and U in lung samples from miners working in coal mines located in the state of Santa Catarina, Brazil. These results were compared to those from a control group constituted of healthy individuals. The results showed that the elements determined exhibit considerable intersubject variability within a single group of individuals and the mean values of concentrations in miners' lungs were higher than those of normal individuals. Lung samples presented U concentrations varying from 11 to 890 micrograms/kg. Therefore, for some samples, the contribution of the uranium fission products in the analysis of La, Ce, Nd, and Sm was considered by determining the interference correction factors. The accuracy of the results was evaluated by analyzing certified reference materials. PMID- 10676504 TI - Environmental control in the uranium mine Lagoa Real, Brazil. AB - Uranium production in Brazil will be restarted in the year 2000, through exploration of the Jazida da Cachoeira, located in the uraniferous region of Lagoa Real, Bahia State, northeast Brazil. Because of the mining operations, an open cast mine of approximately 27 x 10(4) m2 will be produced and the material removed will constitute approximately 14 x 10(6) tons, occupying an estimated area of 50 x 10(4) m2. Currently, there is a new concept about project developing, where the impact assessment is addressed during the initial phases. Beside this, legislation requires an environmental impact assessment before starting mining activities. In this context purpose, it is the purpose of this work to evaluate the chemical composition of the future waste; thus, samples were collected from the rocks that surround the mineral deposits. Instrumental neutron activation analysis was used for the elemental quantification, the mean elemental concentrations were calculated, and the ratios were normalized using North American Shale Composite (NASC). In the raw material eight main mineral structures exist whose elementary composition were compared using the statistical techniques of analysis of variance, ranking ANOVA, and multivariate ANOVA. PMID- 10676505 TI - Comparative study of desert truffles from Kuwait and other countries in the Middle East for radionuclide concentration. AB - Concentrations of some radionuclides, including 137Cs, in desert truffles in Kuwait were studied and compared with similar samples from other countries in the Middle East, namely Iran, Egypt, and Tunisia. In addition, sand samples from Kuwait were assayed to calculate the transfer factor of the radionuclides under consideration. The measured concentrations of 40K, 226Ra, and 137Cs show that 137Cs is much higher in Egyptian samples, whereas 40K is much lower in samples from Tunisia. The average effective dose equivalent calculated for the Kuwaiti population according to their diet habits was found to be in the range 0.14-0.23 microSv/a. The results are compared with values from other countries. PMID- 10676506 TI - Study of environmental biomonitoring of the Jiaozhou Bay. AB - The content of 34 elements was determined in tissue samples of several marine bivalve species collected from various sites in the Jiaozhou Bay. The scope of the study was to determine the most suitable bivalve species to be used for environmental biomonitoring and to evaluate the environmental status of the bay. Clams exhibited higher elemental contents than oysters and they are the major marine bivalve species in the Jiaozhou Bay; therefore, we consider clams to be more suitable than oysters as bioindicators for evaluating the environmental status of the area. Increased elemental levels in clam tissues indicate polluted sites. Also, increased elemental levels in mussels point to possible pollution from tourism development at one selected site. PMID- 10676507 TI - Measurement of fluorine pollutant in plant leaves and soil using nuclear reaction analysis. AB - In this article, the soil and the leaves of plants, parasol, cotton, and glossy privet around a fluorine-polluted area were taken as the samples, and fluorine concentration of the samples were studied using the nuclear reaction 19F(P, alpha)16O, and some results were given. PMID- 10676508 TI - Multielement compositions of marine phytoplankton samples from coastal areas of Japan by instrumental neutron activation analysis. AB - Phytoplankton samples were collected during spring bloom of diatoms from three coastal areas of Japan using a NORPAC P-25 net (25-micron opening) with a NGG52 prenet (335-micron opening), and 25 major and trace elements have been analyzed by INAA. Concentration ranges of analyzed phytoplankton samples are much wider than the concentration ranges compiled by Bowen (1979) except for As, and data of marine phytoplankton samples for Br, Sb, Hf, Sc, La, Ce, Sm, and Eu were not included in the compilation. The 25 analyzed elements have been categorized into three groups: elements showing positive correlation with Br, positive correlation with Al, and no positive correlation with Br or Al. The marine phytoplankton samples have been plotted on a Masuzawa-Koyama-Terazaki (MKT) plot and it proved that the MKT plot is applicable to marine phytoplankton samples. PMID- 10676509 TI - Neutron activation analysis of Se in bovine plasma samples. AB - Elemental concentration of selenium in bovine plasma at parts-per-billion levels was determined using radiochemical neutron activation analysis (RNAA). This study was connected with the relation between the Enteque Seco, a toxic bovine calcinosis, and the Se status. The technique developed is based on the coprecipitation of Se with HgS. Good agreement with certified reference materials have been found. The values for the Se contents in normal bovine samples were in concordance with literature values. PMID- 10676510 TI - Plutonium, 241Am, 90Sr, and 137Cs concentrations in some Antarctic matrices. AB - A radioecological survey in Antarctica shows that the 239 + 240Pu, 238Pu, 241Am, 90Sr, and 137Cs activities were detectable in nearly all the samples. The activity level of 239 + 240Pu, 241Am, and 137Cs in antarctic sediments was about 5-20 times lower than in the northern Adriatic Sea sediments, but the 238Pu activities were relatively high. It was interesting to note that the 90Sr concentrations in all the sediments tended to be low, which could be the result of the easier exchangeable behavior of 90Sr in water. High concentrations were detected in mosses and lichens and their activity levels were comparable to those in central Italy. The radionuclide ratio analyses show that the major part of 239 + 240Pu, 241Am, 90Sr, and 137Cs was a result of nuclear weapon tests. The higher 241Am/239 + 240Pu ratio was observed and it could perhaps be the result of fallout of nuclear weapon tests prior to 1962. The 238Pu/239 + 240Pu ratio in the antarctic matrices was about seven times higher than in the Northern hemisphere and it could be inferred that the major part of 238Pu was originating from the SNAP-9A satellite accident. PMID- 10676511 TI - Bioaccumulation of rare earth elements in cultured HeLa cells. AB - HeLa S-3 cells were grown in minimal essential medium supplemented with 10% calf serum and 1 mM L-glutamine without adding any rare earth elements (REEs). Exponentially growing cells were collected, and dried materials were used to analyze their REE content by inductively coupled plasma-mass spectrometry. The results showed that the cells accumulated REEs in individually different manners; namely the accumulation ratio was higher in the lighter REEs than in the heavier REEs. To deduce the implication of the accumulation of REEs in HeLa cells, the accumulation ratios for REEs were compared with those of other biologically important elements. It was seen that the accumulation ratios obtained for REEs (from 31.8 [Ce] to 14.7 [Lu]) were intermediate among those of many bioelements: Fe (124), Mg (54.5), K (38.8), Cr (12.7), Na (11.8), Mn (11.3), Zn (10.7), Ca (8.8), and V (6.7). PMID- 10676512 TI - Elemental analysis of herbal preparations for traditional medicines by neutron activation analysis with the k0 standardization method. AB - Medicinal herb preparations prescribed for specific treatment purposes were purchased from markets and were analyzed by instrumental neutron activation analysis with k0 standardization. Then, 500-700 mg of each sample was pelletized under a pressure of six tones and irradiated together with monitors for alpha and neutron flux ratio determinations for about 6 h in a thermal flux of 2.29 x 10(12) n/cm2/s. The accuracy of the method was established by analyzing standard reference materials. Twenty-nine elements, Ag, As, Au, Ba, Br, Ca, Ce, Co, Cr, Cs, Eu, Fe, Hf, K, La, Mn, Mo, Na, Rb, Sb, Sc, Se, Sm, Sr, Th, U, Yb, and Zn, were measured in all the samples, and Hg was detected in some samples, with good accuracy and reproducibility. The concentration of elements determined was found to vary depending on the composition of the herbs used. Although the trend linking the element of the medicinal plants to its curative abilities could not be clearly determined, this study showed that the toxic elements found in the samples were below the levels prescribed by health regulations. Nevertheless, such data are important to understand the pharmacological action and the exact mechanisms of action and formation of active constituents for each medicinal plant and to decide the dosage of the herbs used in the final formulation. PMID- 10676513 TI - Determination of selenium in Canadian food items by cyclic instrumental neutron activation analysis. AB - Instrumental neutron activation analysis (INAA) and pseudocyclic INAA (PCINAA) have been used to determine the selenium content of a variety of Canadian food items. Use of the 162-keV gamma ray of short-lived 77mSe in INAA allowed relatively simple and rapid determinations and was suitable for many of the foods. PCINAA was found to give lower detection limits and was used for the low selenium food samples. Both internal and external quality assessments were used to evaluate and assure the accuracy and precision of the methods developed. PMID- 10676514 TI - Lead and copper contamination of soil from industrial activities and firing ranges. AB - Lead still remains one of the most thoroughly investigated heavy metals in the environment. Although the identification of lead in soil is a routine matter, its environmental consequence is still much debated because of its potential mobility. We have investigated lead- and copper-contaminated soil from two different areas. One was in an urban area, which formerly had a lead smelter within the city. The other a firing range, in which hundreds of thousands of rounds were fired into a very large mound known as a berm. Homogeneity tests, depth profiles, and Pb-Cu correlations are discussed. PMID- 10676515 TI - Radiochemical neutron activation analysis in the life sciences. A look back and ahead. AB - The contribution of radiochemical neutron activation analysis (RCNAA) to a better understanding of trace element analytics and physiology in the life sciences is outlined. Now, various non-nuclear powerful techniques for trace element analysis have become available, competing with RCNAA. This necessitates re-evaluation of the position of RCNAA, in particular versus inductively coupled plasma--mass spectrometry (ICP-MS). On basis of the characteristic features of RCNAA and the capabilities of various competing non-nuclear analytical techniques, future niches for RCNAA in the analytical market are indicated. PMID- 10676516 TI - Prompt gamma-ray analysis using cold and thermal guided neutron beams at JAERI. AB - A highly sensitive neutron-induced prompt gamma-ray analysis (PGA) system, usable at both cold and thermal neutron beam guides of JRR-3M, has been constructed. The system was designed to achieve the lowest gamma-ray background by using lithium fluoride tiles as neutron shielding, by placing the samples in a He atmosphere and by using a Ge-bismuth germanate detector system for Compton suppression. The gamma-ray spectrometer can acquire three modes of spectra simultaneously: single, Compton suppression, and pair modes. Because of the low-energy guided neutron beams and the low-background system, analytical sensitivities and detection limits better than those in usual PGA systems have been achieved. Boron and multielemental determination by a comparative standardization have been investigated, and accuracy, precision, and detection limits for the elements in various materials were evaluated. The system has been applied to the determination of B and multielements in samples of various fields such as medical, environmental, and geological sciences. PMID- 10676517 TI - Is loss-free counting under statistical control? AB - A new formula for the statistical uncertainty of "loss-free counting" (LFC) is presented. Its validity is demonstrated by comparing with experimental data obtained with a HPGe gamma-ray spectrometer. Also, computer simulation data of nuclear counting with different types of count loss (pileup rejection, extending and nonextending dead time) are in agreement with the predicted counting uncertainty. The proposed formula for LFC uncertainty is applicable to spectrometers with a classical semi-Gaussian pulse-shaping amplifier as well as with a gated-integrator amplifier. Hence, achieving statistical control seems to be a feasible goal. PMID- 10676518 TI - Limits of detection of a total reflection X-ray fluorescence system with double reflection module. AB - An X-ray tube with a Mo target and Zr filter, operated at 45 kV/20 mA, was used to excite samples (5 microL deposited on a quartz support) and the total reflection angle condition was obtained with a double reflector module built with two 10-cm-long 7-mm-thick quartz crystals placed 50 microns apart. A high resolution spectrometer based on a Si(Li) detector coupled to a multichannel analyzer was used for X-ray detection and the spectra were interpreted with the AXIL software. The system was calibrated with standard chemical solutions containing Cr, Fe, Cu, Zn, and Pb, and Y was used as an internal standard to correct eventual geometric errors and high-voltage instabilities of the X-ray generator. The limits of detection were 19, 9, 5, and 4 ng/mL for Cr, Fe, Cu, and Zn, respectively, analyzed through characteristic K alpha X-rays, and 7 ng/mL for Pb, through L alpha X-rays, considering 50 microL samples deposited and dried on a quartz support, to be excited/detected for 1000 s. PMID- 10676519 TI - Matrix effects in PIXE analysis of aerosols and ashes. AB - A comparison of instrumental neutron activation analysis (INAA) and proton induced X-ray emission (PIXE) results for size-fractionated atmospheric aerosols ("coarse" and "fine" fractions with an equivalent aerodynamic diameter of 2-10 microns and < 2 microns, respectively, or the PM10 fraction) showed that PIXE yielded significantly lower results for the PM10 and coarse fractions, especially for elements with a low Z resulting from a particle size effect. Somewhat lower PIXE results were also obtained for the fine fraction of atmospheric aerosols. A correction is also needed for irregularly shaped deposits of combustion aerosols collected by a cascade impactor in 11 size fractions ranging from 0.016 to 14.3 microns, as well as for thick samples of fly and bottom ashes. An equivalent layer thickness (ELT) model is proposed to correct the matrix effects in PIXE. The approaches for the calculation of ELT using a comparison of PIXE and INAA results or by comparing PIXE results obtained using two different incident proton beam energies (1.31 and 2.35 MeV) are described. The correction for the ash pellets and irregular deposits are also discussed. PMID- 10676520 TI - Comparison of the relative and k0 methods for the standardization of NAA with stable low-flux reactors. AB - The k0 standardization method was adapted for NAA with stable low-flux reactors where flux monitors are not needed. The modified k0 method offers the convenience of the use of libraries of sensitivity constants. It was compared to the relative method for 52 elements using a SLOWPOKE reactor and 6 counting geometries. The sensitivity constants determined from k0 values were found to be as accurate or more accurate than those measured with standards. NAA with this modified k0 method should be accurate to 3% for light elements and 5% for heavy elements. PMID- 10676521 TI - An evaluation of thermal and epithermal neutron activation analysis compton suppression methods for biological reference materials. AB - For neutron activation analysis (NAA), the usual matrix problems of sodium, chlorine, and bromine are well known to give rise to high backgrounds that inhibit the determination of several trace elements for short-lived or medium lived NAA. For long counting times in long-lived NAA, very low backgrounds are required to achieve good sensitivities. We have investigated the use of thermal and epithermal NAA in conjunction with Compton suppression to determine several elements such as arsenic, antimony, cadmium, and mercury, at the level of a few nanograms. The values of these techniques are discussed in contrast to the standard radiochemical methods. PMID- 10676522 TI - Determination of iodide amounts in urine and water by isotope dilution analysis. AB - Urinary iodide and iodine in drinking water were determined in 318 healthy children aged 0 to 18 yr living in Izmir and environmental rural and urban areas in the western part of Turkey. The method is based on substochiometric isotope dilution analysis. Iodide was precipitated by substoichiometric amounts of AgNO3. Iodide-131 was used as a tracer. Electrophoresis was performed to separate Ag131I from excess 131I-. The Ag131I zone was cut off the electrophoresis paper and counted with a NaI(Tl) scintillation counter. Count rates were plotted versus added KI concentrations. The unknown iodide amount was found by using these linear plots. Iodide concentration ranges were within 1.8-100.45 micrograms/L in the analyzed drinking water samples. The mean value was 44.14 +/- 17.33 micrograms/L and the median was 58.08 micrograms/L. Urinary iodide concentration ranges were 0.22-142.22 micrograms/L. The median of the distribution was 37.71 micrograms/L and the mean was 40.30 +/- 24.05 micrograms/L. The results show that the examined area suffers moderate iodine deficiency. PMID- 10676523 TI - On the fission interference correction and its dependence on the epithermal to thermal neutron flux ratio in thermal NAA of molybdenum. AB - The present work aims at the following: (1) analyzing the experimental fission interference factor for molybdenum, FMo, obtained by the authors, who have described the irradiation conditions used as concerns the epithermal to thermal neutron flux ratio, phi epi/phi 0; (2) establishing a simple calculation model that describes the dependence of FMo on phi epi/phi 0 in an adequate way, to provide a satisfactory basis to explain the scatter found in the existing experimental data; and (3) clearly indicating the basic recommendations to take into account in order to obtain with high accuracy the concentration of molybdenum in samples containing uranium. PMID- 10676524 TI - Effects of reactor temperature and sample mass on the activation of biological and geological materials with a SLOWPOKE reactor. AB - Neutron activation analysis with a SLOWPOKE reactor relies on the stability of the neutron flux in the irradiation sites. Flux monitors were irradiated to measure the flux variation with the reactor temperature and with the amount of moderator in the irradiation vial. The thermal flux decreased by 2.7% for a 10 degrees C increase in reactor temperature. The thermal flux increased by up to 8% and the fast flux decreased by up to 13% depending on sample size and hydrogen content. PMID- 10676525 TI - Direct preparation of radioactive fullerenes as a tracer for applications. AB - The C60 and C70 fullerenes were irradiated by high-energy gamma-rays and charged particles. The irradiated samples were dissolved in CS2 and/or toluene and filtered to remove insoluble by-products. Finally, radioactive fullerenes and products labeled with 11C or 13N were isolated and detected in the liquid phase by radiochromatography. It was found that (1) not only 11C or 13N radioactive monomer fullerenes but also their dimers (trimers and, possibly, tetramers) were produced by recoil implantation process following nuclear reaction and (2) the radioactive fullerene labeled with 11C yields has led to high yields. PMID- 10676526 TI - Effect of air cavities on the dose delivered to the lung during high-dose brachytherapy. AB - In the treatment of lung cancer using the radiotherapy technique of intracavitary brachytherapy with an 192Ir source, the lung is normally assumed to be entirely composed of a homogeneous mass of soft tissue. The aim of this study is to investigate whether there is the possibility that the air cavities in the lung influence the dose delivered to the lung at a prescribed distance from the source. The Monte Carlo code MCNP-4A was used to model the dose delivered by both 192Ir and 198Au as a function of treatment medium, density and composition, photon energy, and distance from the source. The suitability of MCNP-4A for this study was tested by producing depth-dose profiles for photons in water and comparing these to calculated profiles produced using well-documented methods. PMID- 10676527 TI - Tritium incorporation into functional materials by applying OT-for-OH exchange reaction. AB - In order to reveal (1) the behavior of the tritium-labeled hydroxyl group (i.e., OT group) and (2) the effect of temperature on the dissociation equilibrium of tritiated water, the OT-for-OH exchange reaction in a homogeneous system was observed (1) using ion-exchange resins and HTO water, and (2) using hydrogen oxides and HTO water. Consequently, the following four conclusions were obtained: (1) The OT-for-OH exchange reaction occurred between each anion-exchange resin (or each metal hydroxide) and HTO water. (2) The higher the temperature is, the smaller the activity of both anion-exchange resin and metal hydroxide will be, and the activity of cation-exchange resin is large when the temperature is high. (3) The larger the degree of crosslinking in each Biorad AG1 resin is, the lower the activity of the resin will be. (4) The exchange rate for the OT-for-OH exchange reaction is small when the electronegativity of the metal ion in the metal hydroxide is large. PMID- 10676528 TI - Experience on the neutron activation of natural/enriched Re, Sm, and Ho nuclides in a reactor for the production of radiotherapeutic radionuclides. AB - In recent years, much effort has been concentrated on the use of beta-emitting radionuclides for the treatment of various cancers. The reports suggested the application of 186Re and 153Sm as radiotherapeutic radionuclides for the treatment of palliative widespread skeletal metastases, whereas 166Ho was suggested as an agent for radiation synovectomy. Hence, a study on the production of 186Re, 153Sm, and 166Ho radionuclides was carried out by neutron activation of the appropriate target materials using a Pakistan Atomic Research Reactor (PARR 1) at a neutrons flux of 1 x 10(4) n/cm2 s. These radionuclides were then converted to appropriate radiopharmaceuticals for their use on animals and patients. The targets of natural Re (metal), natural Sm2O3, enriched Sm2O3 (99.06%), Sm(NO3)3 (solid), Sm(NO3)3 (liquid), and Ho2O3 were irradiated in the PARR-1. After irradiation, the purity of these radionuclides were checked by a multichannel analyzer, Canberra series 85 (MCA) coupled with HPGe detector and then measured in radioisotope calibrator Capintec ionization chamber model CRC 5RH. The effect of the irradiation time and amount of target material was investigated on the production yields of the radionuclides. The results showed an increase in the specific activity of Re with an increase in the irradiation time from 1 to 72 h, whereas a decrease in the specific activity was observed with increase in the amount of Re from 10 to 100 mg. Similar results were obtained for 153Sm and 166Ho radionuclides. The results further indicated that the specific activity of powder target was much less than the liquid targets for 153Sm. Their conversion to the appropriate radiotherapeutic radiopharmaceuticals were also carried out by investigating the experimental conditions and acceptable quality of 186Re-HEDP and 153Sm-EDTMP complexes were prepared. These complexes were then used on animals and patients which showed good performance. PMID- 10676529 TI - Determination of trace elements in porcine brain by inductively coupled plasma mass spectrometry, electrothermal atomic absorption spectrometry, and instrumental neutron activation analysis. AB - Methods have been developed for the analyses of trace metals in various areas of porcine brains, (temporal, parietal, frontal cortex, both right and left hemispheres). Determinations were carried out using inductively coupled plasma mass spectrometry (ICP-MS) and electrothermal atomic absorption spectrometry (ETAAS). The elements investigated were Li, Mn, Cu, Zn, Cd, Hg, and Pb by ICP-MS and Cu, Cd, and Mn by ETAAS. For determination by ICP-MS, a method of standard additions calibration coupled with internal standards was used, and for ETAAS, standard additions calibrations were prepared. The accuracy of all methods was determined using NIST and IAEA certified reference material. A small number of pig brains were analyzed by instrumental neutron activation analysis for Cr, Co, Cs, Fe, Rb, Se, Sc, Sb, and Zn using the comparator method of analysis. Four separate NIST standard reference materials have been used to examine the validity of the comparator method. PMID- 10676530 TI - A comparative study in Alzheimer's and normal brains of trace element distribution using PIXE and INA analyses and glucose metabolism by positron emission tomography. AB - The onset of Alzheimer's disease has been shown to affect trace element concentrations in the brain when compared to "normal" subjects in ex vivo samples. The techniques used to determine trace element concentrations were proton-induced X-ray emission and instrumental neutron activation analysis. With these methods, significant differences are seen between lobes within a hemisphere and between the same lobes of opposing hemispheres for "normal" brains. The change observed in trace element concentrations may indicate a possible alteration in the function of the blood-brain barrier, the effect of which can be investigated in vivo using the imaging technique of positron emission tomography (PET). A PET study was performed on nine female and nine male subjects to determine whether the regional metabolic rate of glucose (rCMRGlu) varied between hemispheres and sex in the Alzheimer diseased brain as was seen in the trace element study. Glucose metabolism was measured using [F-18]-fluorodeoxyglucose (18F-FDG). Hemispherical differences were observed for the frontal, occipital, parietal lobes, and the temporolateral region in both males and females for rCMRGlu. Variation was also seen between sexes, where the frontal lobe had a lower rCMRGlu in females compared to that of males. PMID- 10676531 TI - Accumulated experience with prenatal diagnosis of Menkes disease by neutron activation analysis of chorionic villi specimens. AB - Since 1983, prenatal diagnosis of Menkes disease has been carried out by determining Cu in samples of chorionic villi from the fetus by means of radiochemical neutron activation analysis. Concentrations of Cu in chorionic villi from male fetuses later confirmed to have Menkes disease were invariably higher than previously reported values for normal controls. Out of 240 samples analyzed in the period 1983-1998, there were 71 from female fetuses that could be carriers of the Menkes genetic defect without suffering from the disease. Increased concentrations of Cu in these samples could not be attributed to the presence of this genetic defect, but might result from sporadic contamination of the samples before analysis. Such contamination also may occur in samples from male fetuses and thus raise the level of Cu in small, but normal specimens into the range characteristic of Menkes disease. In spite of a strict protocol for taking samples without contamination, a total of four false positives were reported during the period of investigation; no false negatives have occurred. PMID- 10676532 TI - INAA of trace elements in colorectal cancer patients. AB - The concentration of trace elements in samples of both colorectal cancer tumors and normal tissues of a Mexican population were irradiated for 30 s and 4 h and their elemental content were measured by instrumental neutron activation analysis. Ca, Cu, Co, I, Mg, Se, Fe, Zn, Hg, Ba, and Cr were analyzed. Alterations in Co, Fe, I, and Ba were found in tumors with respect to normal tissues. PMID- 10676533 TI - Determination of trace elements in tissue of human uterine cancer by instrumental neutron activation analysis. AB - In this article, the low-temperature freeze-drying pretreated technique and instrumental neutron activation analysis were used to determine 29 trace elements in samples of human uterine cancer tissue. The content of these trace elements in uterine cancer tissue was compared with that in cervicitis tissue and in healthy tissue, respectively. Preliminary results indicated that significant differences in contents of Au, I, and Se were observed in these tissues. PMID- 10676534 TI - Elemental composition changes between breast tissue with and without silicone gel sheeting and hypertrophic scar tissue. AB - Hypertrophic scars occur after dermal trauma and are characterized by being elevated above normal skin level as a result of an abundance of collagen. The application of silicone gel sheeting (SGS) has been found to be an effective method of treatment, causing them to regress much quicker than they would do naturally. Normal skin and hypertrophic scar tissue were characterized using proton-induced X-ray emission (PIXE). Skin tissue that had been covered in SGS was also analyzed. For each element and sample type, the concentrations in the epidermis were plotted against the dermis. By considering the concentrations of breast tissue with and without SGS, it could be seen if the SGS changed the compositional structure of the skin. It was found that for the elements P, S, Cl, and K the SGS has no effect on the structure of the skin, as both breast types (with and without SGS) have regression lines that overlap. However, this work shows that there are significant differences for P in the dermis and Cl in the epidermis between the breast tissue with SGS and its control. Therefore, this work shows that the effect the SGS has on concentration occurs similarly for both the epidermis and dermis. PMID- 10676535 TI - Cell-killing efficiency and number of platinum atoms binding to DNA, RNA, and protein molecules of HeLa cells treated with combinations of hyperthermia and cis diamine(glycolato)platinum(II). AB - HeLa S-3 cells were treated with 195mPt-radiolabeled cis diamine(glylato)platinum(II) (254-S) for 60 min at various temperatures, and the relationship between the lethal effect and the number of Pt atoms binding to DNA, RNA, and proteins was examined. The mean lethal concentration (D0) of 254-S for a 60-min treatment at 0 degree C, 25 degrees C, 37 degrees C, 40 degrees C, 42 degrees C, and 44 degrees C was 233, 132, 61.1, 42.7, 25.6, and 9.9 microM, respectively. By using identically treated cells, the numbers of Pt atoms combined with DNA, RNA, and protein molecules were determined in the subcellular fractions. Thus, the D0 values given as drug concentrations were replaced with the number of Pt atoms combined in each fraction. The, the cell-killing efficiency of the Pt atom was expressed as the reciprocal of the number of Pt atoms combined and was calculated for each molecule. The efficiency for the DNA molecule was 0.61 x 10(4), 1.09 x 10(4), 1.88 x 10(4), 1.90 x 10(4), 2.66 x 10(4), and 5.88 x 10(4) nucleotides, respectively, for the conditions described. From 0 degree C to 44 degrees C, the cell-killing efficiency of Pt atoms increased by a factor of 9.6. PMID- 10676536 TI - Determination of metallic ion transfer from an implanted prosthesis by the PIXE method. AB - Prostheses can release some metallic elements to the surrounding tissues, particularly when they are not covered with a biomaterial layer and when an unsealing process happens. We try to measure major and trace elements in these tissues with an experimentally sensitive method. Proton-induced X-ray emission is used to detect about 10 elements in tissue. Tissues are calcinated and deposited in a thin layer before irradiation. Results are obtained in a standard and samples from three patients. We observe contamination by Ti, Cr, Ni, and Zn in the tissues. Correlations are to be studied between these atomic transfers and prosthesis in the patient. PMID- 10676537 TI - Hydrophilic crosslinked copolymers as tissue-equivalent materials for breast cancer detection. AB - Crosslinked hydrophilic copolymers have the potential to be used as breast phantom materials because they can be made to have similar elemental composition to that of body soft tissues. The copolymer, which consists of a combination of hydrophobic monomers (methyl methacrylate [MMA]) and hydrophilic monomers (vinylpyrolidone [VP]), have all the required major elements, such as hydrogen, carbon, nitrogen, and oxygen, found in tissues. Photon attenuation measurements were performed on the crosslinked hydrophilic copolymers in order to ascertain whether they are good representatives of breast tissues in the photon energy range of interest. The results of the measurements of transmission of photons by the hydrophilic copolymers at different discrete energies between 10 and 60 keV are presented. PMID- 10676538 TI - Elemental analysis of blood of Nigerian hypertensive subjects. AB - Proton-induced X-ray emission (PIXE) has been used to obtain the concentrations of 11 elements (P, S, Cl, K, Ca, Fe, Cu, Zn, Br, Rb, and Cd) in whole-blood samples of 16 hypertensive subjects (mean age: 52.5 +/- 0.5 yr) and 18 age matched controls (mean age: 51.5 +/- 0.5 yr) in a Nigerian population. The results of the study indicate that the hypertensive subjects have significantly higher mean concentration of Cl, Cd, Cu, and Zn when compared with the controls, and the mean concentration of P, K, and Ca was found to be significantly lower in the hypertensive group in comparison to the controls. Furthermore, the Zn:Cd ratio was found to be significantly higher in the controls than in the hypertensives, and the Cu:Zn ratio was significantly higher in the hypertensives. PMID- 10676539 TI - Determination of zinc contents in rabbits with cerebral ischemia by NAA and ICP AES. AB - Cerebral ischemia is an important cerebral vascular disease, and zinc is a necessary trace element for humans. In this work, a cerebral ischemia model of rabbit was established by operation. The samples of brain and serum in the animal models were collected. The Zn contents in the samples were determined by neutron activation analysis and inductively coupled plasma-atomic emission spectrometry. The results show the Zn contents in brain decreased 2 mo after cerebral ischemia, and Zn contents in serum decreased even more obviously. In addition, a positive correlation of Zn contents between left and right cerebral hemispheres was observed, and the positive correlation between brain and serum was also observed. A test of Chinese medicine was also carried out in the work. Two Chinese medicines were fed to rabbits with cerebral ischemia in the experiments. The results showed they probably can prevent the decrease of Zn contents in serum. PMID- 10676540 TI - Preliminary study on the relationship between osteoporosis and trace elements with rat models. AB - Thirty rats were divided randomly into five groups and fed with different feeds. They were killed after 2 mo. Right thighs of these rats were taken as samples. Bone mineral densities of these samples were measured by the double-energy x-ray bone mineral densitometer, and trace elements contents of the samples were analyzed by neutron activation analysis. Results of the experiments showed that some elements were well correlated with others, and there were obvious variances between some elements of the normal group and those of other four groups. PMID- 10676541 TI - Study of implanted biomaterial functionality by diphosphonate molecules labeled with radioactive 99mTc. AB - An implanted biomaterial can be transformed into young bone after some months, but it has not necessary reached full biofunctionality. Mineral concentration kinetics and crystal-structure studies, still being carried out in our group, are completed here by biofunctionality determinations. A natural coral is implanted in vivo at the cortical level of the femoral diaphyoff++ in rabbits. Diphosphonates molecules labeled with radioactive 99mTc are then injected in rabbits and the fixation of the radioactivity is analyzed in several sites for 8 mo after the implantation. Nuclear instruments and methods are used for the measurements. Four successive cycles of osseous remodeling are determined before reaching a biofunctional phase. PMID- 10676542 TI - Proximal impact of two first-grade preventive interventions on the early risk behaviors for later substance abuse, depression, and antisocial behavior. AB - We assessed the immediate effects of two universal, first-grade preventive interventions on the proximal targets of poor achievement, concentration problems, aggression, and shy behaviors, known early risk behaviors for later substance use/abuse, affective disorder, and conduct disorder. The classroom centered (CC) intervention was designed to reduce these early risk behaviors by enhancing teachers' behavior management and instructional skills, whereas the family-school partnership (FSP) intervention was aimed at improving parent teacher communication and parental teaching and child behavior management strategies. Over the course of first and second grades, the CC intervention yielded the greatest degree of impact on its proximal targets, whereas the FSP's impact was somewhat less. The effects were influenced by gender and by preintervention levels of risk. Analyses of implementation measures demonstrated that greater fidelity to the intervention protocols was associated with greater impact on behavior ratings and on achievement scores, thus providing some evidence of specificity in the effect of the interventions. PMID- 10676543 TI - Personal resources and the social distribution of depression. AB - This paper addresses the hypothesis that gender, age, marital status, and SES matter for depression partly because of associated differences in the availability and/or impact of the personal resources of mastery and self-esteem. It is argued that findings indicating that the social distributions of these resources complement those for depression would provide preliminary support for this hypothesis. Based on a large urban community sample (n = 1,390), our findings fail to support the availability hypothesis in relation to marital status, provide only modest support in reference to age and gender, but yield compelling support in relation to socioeconomic status (SES). Indeed, variations in the availability of these resources, especially mastery, provide a largely, if not entirely, adequate explanation for the SES-depressive symptoms relationship and accounts for nearly half of the SES-Major Depressive Disorder relationship. Although the significance of mastery was more pronounced among women and unmarried persons, such differences did not contribute to understanding observed gender or marital status variations in depression. PMID- 10676544 TI - Principles for designing randomized preventive trials in mental health: an emerging developmental epidemiology paradigm. AB - An emerging population-based paradigm is now being used to guide the design of preventive trials used to test developmental models. We discuss elements of the designs of several ongoing randomized preventive trials involving reduction of risk for children of divorce, for children who exhibit behavioral or learning problems, and for children whose parents are being treated for depression. To test developmental models using this paradigm, we introduce three classes of design issues: design for prerandomization, design for intervention, and design for postintervention. For each of these areas, we present quantitative results from power calculations. Both scientific and cost implications of these power calculations are discussed in terms of variation among subjects on preintervention measures, unit of intervention, assignment, balancing, number of pretest and posttest measures, and the examination of moderation effects. PMID- 10676545 TI - Opening the black box: using process evaluation measures to assess implementation and theory building. AB - The past decade has seen increasing recognition in prevention science of the need to move away from a black box approach to intervention evaluation and toward an approach that can elaborate on the mechanisms through which changes in the outcomes operate (Chen & Rossi, 1989; Durlak & Wells, 1997; Spoth et al., 1995). An approach that examines issues of program implementation is particularly critical in the design of efficacy studies of school-based preventive interventions. Numerous preventive intervention strategies are now delivered within the schools, often by regular classroom teachers. The extent to which teachers faithfully deliver a particular curriculum or incorporate instructional strategies emphasized by an intervention is a critical question for the overall project evaluation. This article illustrates the utilization of process measures from a multicomponent school-based prevention program to examine implementation of a teaching staff development intervention, and the program's underlying theoretical basis. Given the nested study design, the analyses utilize hierarchical linear models (Bryk & Raudenbush, 1992) to examine changes in teaching strategies by condition and investigate the hypothesized relationships between teaching practices and student behaviors based on the program's theoretical framework. Results suggest that teaching practices in two of the six intervention focus areas were positively impacted in the first 18 months of the project. Findings also support the relationships between teachers' instructional practices and students' behavior. PMID- 10676546 TI - Reflections on oncology in central and eastern Europe. AB - In the last two decades cancer mortality dramatically increased in the majority of Central and Eastern European countries. In the period from 1992 to 1995, the Hungarian male population had the highest death rate (265.0 of 100,000) due to malignancies among 46 countries worldwide. Hungarian women ranked third in cancer death rate among these countries (138.0 of 100,000). Several factors might be responsible for these figures: a) increases in environmental carcinogenic risk factors, b) unfavourable lifestyle changes in the population especially related to high tobacco consumption and dangerous drinking habits, c) lack or insufficiency detection, d) delay in diagnosis, e) inadequate therapeutic patient management, f) shortage of manpower, and g) unsatisfactory financial support. Efforts have been made to overcome these difficulties by: a) detail analysis of exogenous risk factors, b) review of lifestyle of the population, c) public education efforts for effective prevention, d) introduction of model screening programs, e) reorganization of cancer diagnosis and treatment services, and f) design and establishment of a National Cancer Control Program. PMID- 10676547 TI - The European Society for Medical Oncology (ESMO) and its activities through the Central Eastern European Task Force. AB - The article describes the history and organization of the European Society for Medical Oncology. The society, founded in 1975, aims at advancing medical oncology on a pan-European basis. Postgraduate training and education constitute a major part of ESMO's activities through a current CME programme of courses and other activities. Each year the ESMO Examination is held, and every other year ESMO organizes its congresses with the latest attendance of more than 6000 delegates. ESMO has a continuous increase of members, also from outside Europe. In 1996 ESMO created the Central Eastern European Programme with the aim to support the needs of the countries of the former Eastern Europe. A task force (CEE TF) with members from 16 Central Eastern European countries meets twice a year to discuss key areas. An e-mail communication system has been launched, courses are planned for 1998-1999, exchange programmes are in progress, and support in setting up national guidelines will follow. A Central Eastern European Oncology Group (CEE OG), which performs clinical trials on a cooperative basis, has been established with ESMO guidance. PMID- 10676548 TI - The pharmaceutical industry and oncology in central and eastern Europe. AB - Major opportunities exist for patients, investigators and the pharmaceutical industry in oncology drug development in Central and Eastern Europe. Novel therapeutics may be offered for investigational use in selected centres capable of adherence to Good Clinical Practice (GCP). Requirements for participation in oncology clinical trials include the availability of experienced qualified investigators highly motivated to conform with the principles of GCP (International Harmonization (ICH) guidelines); availability of appropriate Institutional Review Board for Human Subjects (IRB), access to appropriate patient populations, access to individual patient data, acceptance of possible audit by sponsoring companies and the Food and Drug Administration (FDA), and a willingness to participate in the generation of new knowledge. Patients gain through access to novel therapeutics. We have had success in performing clinical trials to international standards in Central and Eastern Europe. This experience will be described. PMID- 10676549 TI - Progress in the non-Hodgkin's lymphomas. PMID- 10676550 TI - Treatment of Hodgkin's disease: current strategies of the German Hodgkin's Lymphoma Study Group. AB - At present over 90% of early stage Hodgkin's disease patients will be cured. Both radiotherapy and combination chemotherapy are effective treatment modalities. However, the optimal choice of treatment or combinations of treatment is still debated. Recently, several trials reported excellent treatment results with combined modality in early stages of Hodgkin's disease. The use of chemotherapy regimen not including alkylating agents may avoid the risk of infertility and secondary malignancies and facilitates reduction of dose and field size of radiotherapy in early stages. In intermediate stages new chemotherapy regimen (i.e., BEACOPP) will offer the chance to reduce the fraction of patients with initial treatment failure, while reducing the extent of radiotherapy. With the introduction of the escalated BEACOPP regiment it was demonstrated that the prognosis of the advanced stages could be positively influenced by intensification of therapy. Future trials aim to answer: 1) which chemotherapy regimen in which quantity will be the best with respect to efficacy and longterm toxicity and 2) which dose and field size of radiotherapy is adequate within the combined modality approach. PMID- 10676551 TI - Multiple myeloma: an update on biology and treatment. AB - Recently, several advances have been made in understanding the pathogenesis of multiple myeloma. Increasing evidence favours a pre-switched, but somatically mutated B-cell as myeloma stem cell to give rise to the malignant clone. Deletions of the p53-gene, partial or total loss of chromosome 13 and rearrangements of band 14q32 and 11q13 are frequently found in multiple myeloma, and were shown to harbour prognostic significance. Presence or absence of distinct chromosomal aberrations may guide selection of treatment strategies in the future. Although melphalan/prednisolone remains the standard of myeloma treatment in elderly patients, significant improvement has been achieved in antimyeloma and in supportive therapy. High dose therapy with autologous stem cell transplantation enhances survival in younger patients and several trials are ongoing to substantiate these results. The effects of interferon maintenance treatment on overall survival is significant in metaanalysis, although the gain achieved is limited. Newer treatment strategies--targeting the molecular level- have just entered clinical trials, and may further improve outcome of myeloma patients. PMID- 10676552 TI - Treatment of acute leukemia. AB - Leukemic cells are highly sensitive to chemotherapeutic agents. A reduction of the leukemic burden is easily achieved by chemotherapy in most cases. However, it is difficult to reduce the number of leukemic cells to such an extent that a regrowth does not occur and the patient is cured. Traditionally the therapy of acute leukemia is divided into induction and post remission therapy. The aim of the induction therapy is to reduce the number of leukemic cells to a morphologically undetectable level allowing normal hemopoiesis to recover. The goal of the post remission treatment is a further reduction of leukemic cells to zero or to very low levels which can be controlled by (still unknown) endogenous mechanisms. In some recent treatment protocols induction and the early part of post remission treatment are not strictly separated. PMID- 10676553 TI - Modulation of multidrug resistance (MDR) in hematological malignancies. AB - The term multidrug resistance (MDR) describes the observation that tumour cell lines can become cross-resistant to several structurally unrelated chemotherapeutic agents after exposure to a single cytotoxic drug. In hematological malignancies, MDR is most often associated with overexpression of P gp, a 170-kd transmembrane glycoprotein encoded by the human MDRI gene. Indeed, P gp expression has been correlated with drug sensitivity and clinical outcome in several studies in acute myelogenous leukemia (AML), multiple myeloma (MM), and malignant lymphomas (NHL). A large number of compounds 'off the shelf' have been investigated for their ability to reverse the P-gp mediated MDR. However, most of these agents produced severe toxic effects at doses required to effectively block P-gp function, and modulation of P-gp in normal tissues can affect the pharmacokinetics and, thus, the toxicity of the associated chemotherapeutic agents. Phase I/IIa trials with third generation MDR modulators, such as valspodar, show that these new agents can be safely administered in combination with different chemotherapy regiments after dose adjustments of cytotoxic drugs that a P-gp substrates. Moreover, MDR reversal by valspodar has been demonstrated in the patients with AML, multiple myeloma, and non-Hodgkin's lymphoma. The definition of the clinical benefits of using MDR modulators in haematological malignancies and their full extent awaits the conclusion of the ongoing randomized phase III trials with valspondar in either newly diagnosed or resistant relapsed AML patients, and in multiple myeloma patients who have failed front-line treatment. PMID- 10676554 TI - The role of energy and fat in cancers of the breast and colon-rectum in a southern European population. AB - BACKGROUND: Several uncertainties remain with respect to the role of intake of fat and/or total energy in the etiology of cancer of the breast and colon-rectum. PATIENTS AND METHODS: Between 1991 and 1996, 2569 women with incident breast cancer (median age: 55 years), 1953 subjects with cancer of the colon-rectum (median age = 62), and 5155 hospital controls were interviewed in six Italian areas. The validated food frequency questionnaire included questions on 78 foods and recipes and specific questions on individual fat intake pattern. RESULTS: Significant trends of increasing breast and colorectal cancer risk with increasing intake emerged for bread and pasta, pork and processed meats and potatoes (breast cancer only), cakes and desserts (colon-rectum cancers only), and refined sugar. Most vegetables were inversely associated with cancer of the colon and rectum, whereas only carrots and raw vegetables seemed to lower breast cancer risk. High fruit intake was associated only with a reduction of rectal cancer. Total energy intake was directly associated with all cancer sites. Among macronutrients, high intake of starch and saturated fat seemed to lead to an increase of cancer risk. High intakes of polyunsaturated fatty acids (chiefly derived from olive oil and seed oils) were protective. Among micronutrients, beta carotene, vitamin E, and calcium showed inverse associations with breast and colorectal cancer risk. CONCLUSIONS: An excess of energy intake, particularly from refined bread and pasta, can be an unfavourable feature of the Mediterranean diet, in the presence of a sedentary lifestyle. PMID- 10676555 TI - Once-only sigmoidoscopy. AB - The rationale for infrequent screening for colorectal cancers by sigmoidoscopy of the general population around age 60 is reviewed. A progress report for a trial to evaluate this approach is also presented. PMID- 10676556 TI - Adjuvant and induction chemotherapy in non-small cell lung cancer. AB - About 25%-30% of patients with non-small cell lung cancer can be resected with curative intent. However, systemic relapses occur in up to 70% of these patients. Thus, postoperative adjuvant chemotherapy was evaluated in several randomised trials but the results of these trials were inconclusive with a survival benefit only in some trials. Shortcomings of these trials included low number of patients, poor patient compliance and inadequate chemotherapy protocols. A recent meta-analysis suggested an absolute survival benefit of 5% at five years for postoperative cisplatin-based chemotherapy as compared to surgery alone. Thus adjuvant chemotherapy with both improved chemotherapy protocols and improved anti emetics is currently re-evaluated in several randomised trials on large patient populations. PMID- 10676557 TI - Chemotherapy of advanced non-small cell lung cancer. AB - Until recently the role of chemotherapy in NSCLC has generally been questioned. Major concerns included marginal activity, considerable toxicity and high cost of this treatment. There has, however, been increasing evidence from individual studies and meta-analyses that chemotherapy in advanced NSCLC is able to increase survival and improve quality of life. In the past few years a series of active drugs (paclitaxel, docetaxel, gemcitabine, vinorelbine, topotecan and irinotecan) with novel mechanisms of action and favourable toxicity profiles have been developed. These agents appear to hold the promise of added therapeutic benefit. In consequence, chemotherapy has currently been considered an important part of the standard treatment in selected patients with advanced NSCLC. Despite recent developments, treatment outcomes in advanced NSCLC remain far from satisfactory, and new effective means are desperately needed if more patients are to enjoy the prospects of long-term survival. PMID- 10676558 TI - Treatment of small cell lung cancer patients. AB - Small cell lung cancers, comprising approximately 20% of lung cancers, are rapidly growing and disseminating carcinomas which are initially chemosensitive but acquire drug resistance during the course of disease. Thus, outcome is poor with median survival of 10-16 months for patients with limited and 7-11 months for patients with extensive disease. Polychemotherapy with established drugs (platins, etoposide, anthracyclines, cyclophosphamide, ifosfamide and Vinca alkaloids) plays the major role in the treatment of this disease and results in overall response rates between 80%-95% for limited disease and 60%-80% for extensive disease. Dose-intensified chemotherapy and high-dose chemotherapy with peripheral blood progenitor cell support were tested in several trials but their exact impact on outcome remains to be determined. New drugs including the taxanes (paclitaxel, docetaxel), the topoisomerase I inhibitors (topotecan, irinotecan), vinorelbine and gemcitabine are currently evaluated in clinical trials. In limited disease, thoracic radiotherapy improves survival and prophylactic cranial irradiation should be administered to those with a reasonable chance of cure. PMID- 10676559 TI - Combined modality therapy of non-small cell lung cancers. AB - Lung cancer represents the leading cause of cancer mortality. Non-small cell lung cancer (NSCLC) accounts for about 75% to 80% of lung cancer cases and carries a 5 year survival of about 10% to 15% for all stages. Approximately one third of NSCLC patients present with stage III disease, which is defined as locally advanced tumour confined to the chest without distant metastasis. The traditional treatment for stage III patients has been thoracic radiotherapy (RT). However, the impact of thoracic RT alone has been minimal with published studies showing median survival < 1 year and 5-year survival of 5% to 7%. Thus, the treatment of stage III NSCLC remains a significant challenge. The metastatic nature of this disease has been responsible for the poor survival statistics and emphasises the need for effective systemic treatment. In recent years, cisplatin-containing combination chemotherapy has emerged as a viable option in the treatment of NSCLC. Combined modality therapy employing systemic (chemotherapy) and local (RT with or without surgery) approaches has shown favourable results in patients with stage III disease. Randomised studies have demonstrated the benefit of concurrent or sequential chemoradiation in selected patients with a good performance status and minimal weight loss. The exact sequence has yet to be determined. Moreover, randomised studies in stage IIIA potentially resectable disease show survival advantage for patients receiving combined modality treatment. Thus, combined modality treatment has the potential to improve overall survival by increasing both local and distal control. These recent reports of randomised clinical trials of combined modality therapy for stage III NSCLC form the basis for this report. Several new agents, like the taxanes, CPT-II and gemcitabine show promising activity in NSCLC treatment. Ongoing studies are evaluating the potential role of these new agents in combined modality treatment but since the phase III trials have not been reported yet these studies will not be discussed. PMID- 10676560 TI - Combined modality therapy of rectal cancers. AB - Rectal cancer accounts for about 10% of new cancer cases each year. It strikes men and women at nearly the same rate, generally in the range of 50-80 years of age, with rising incidence with age. Despite simple screening procedures rectal cancer is often advanced when discovered. Current trends in the management of cancer have focused on organ preservation and improved quality of life without compromising the overall survival. During the last decade substantial progress has been made in treatment modalities: new and improved radiation techniques (conformal radiotherapy, altered fractionation, brachytherapy), chemotherapy (protracted infusion, use of radiosensitizers) and development of surgical procedures-enabling safer postoperative irradiation. In patients with advanced/unresectable disease aggressive combined chemoradiation can be added prior to surgery to downstage the tumour and increase the proportion treated with anal-rectal-sparing procedures. Preoperative chemoradiation therapy regimens are as safe and tolerable as the standard postoperative treatment. In this presentation indications for preoperative radiochemotherapy will be discussed in detail, together with treatment-related side effects, prognostic parameters, tumour response and outcome. Different irradiation settings and chemotherapy schedules are described. In patients with primary resectable disease (mainly Dukes C) several prospective randomised trials have shown less local recurrence with postoperative combined modality therapy. PMID- 10676561 TI - New possibilities in chemotherapy for colorectal cancer. PMID- 10676562 TI - BRCA1 and BRCA2 and breast cancer incidence: a review. AB - BACKGROUND: Epidemiological studies have repeatedly shown that having a family history is a risk factor for female (and male) breast cancer. Some rare families have many (4 or more) cases of early onset breast cancer (some of which also include women with ovarian cancer) which are most clearly explained by an autosomal dominant gene with high penetrance. DESIGN: Families with multiple cases of early onset breast (and/or ovarian cancer) have been studied using linkage analysis with the intention of finding the chromosomal region containing such genes. RESULTS: Two predisposition genes, BRCA1 and BRCA2, have been mapped and cloned. Mutations in these genes confer increased risk of cancer, although the precise level of the increased risk is still unclear. The majority of families with four or more cases of breast cancer diagnosed under the age of 60 years are due to mutations in BRCA1 or BRCA2. CONCLUSIONS: The importance of these two genes to familial breast cancer and to breast cancer incidence overall is becoming clearer; the current information is reviewed. The findings can be immediately translated into clinical practice for these multiple case families. The identification of such families raises a number of other important clinical questions concerning patient management. PMID- 10676563 TI - Screening mammography for early detection of breast cancer. AB - From numerous studies on breast cancer it can be concluded that no single measure can lessen the burden of this frequent cancer in women in all developed countries. Complex strategies including primary prevention by identification of risk factors and their modification, secondary prevention by earlier detection and tertiary prevention by improving treatment outcome are needed to control the disease. Besides age, the established breast cancer risk factors include certain benign breast diseases, family history, ionising radiation, some reproductive factors and obesity. Primary prevention includes general recommendation for healthy lifestyle, e.g., avoidance of obesity, proper diet, physical activity and moderate alcohol consumption. Randomised controlled trials conducted in the USA, Canada, Scotland and Sweden have shown that regular mammography, alone or in combination with clinical examination, is effective in reducing mortality for about 30% in women over the age of 50, and much less in younger population. However, mammography screening has several drawbacks, the major being its tendency towards false positive and false negative results with all their potential psychosocial consequences. High quality assurance and control, as well as effective and readily available treatment, all of which demand high investments, are indispensable for good results. Even in the absence of organised screening, the availability of effective treatment may contribute to reduction in breast cancer mortality. PMID- 10676564 TI - Adjuvant therapy of breast cancer: update. AB - The theoretical prediction that breast cancer is a systemic disease, and that patients may benefit from addition of systemic therapy to local treatment, has now been confirmed by three decades of clinical investigations. A long-term follow up of individual trials and the International Overview based on meta analyses clearly showed the potential of both hormonal therapies and chemotherapy to prolong disease-free and overall survival in nearly all groups of patients. The benefits have been demonstrated for both premenopausal and postmenopausal patients, with both node-negative and node-positive disease. However, there is still considerable uncertainty regarding the most appropriate treatment for each individual patient. In the present review, the results of meta-analysis are highlighted in the context of the new trials supporting the value of chemoendocrine therapy and anthracycline-based therapy. The results of prospective randomised trials evaluating the role of dose intensification, drug sequencing and dose density are discussed. Also presented are new treatment strategies, such as preoperative chemotherapy and high-dose chemotherapy with stem cell support, the value of which remains to be confirmed. Future possibilities opened by inclusion of biologics into adjuvant therapy are discussed. PMID- 10676565 TI - New developments in chemotherapy of advanced breast cancer. AB - Anthracyclines and taxanes are the two most active classes of chemotherapy for the treatment of advanced breast cancer. Recent studies have investigated combination therapy including doxorubicin (Dox) and paclitaxel. The efficacy of this combination has been established in a phase III study conducted by ECOG, comparing Dox/paclitaxel versus Dox versus paclitaxel. The combination is superior to Dox or paclitaxel with respect to response rate and time to disease progression, indicating that the combination provides a new standard for the first line treatment of metastatic breast cancer [1]. Phase II studies using higher doses of Dox and using shorter infusions of paclitaxel have suggested the combination can be further optimized; Gianni reported a 94% objective response rate using Dox 60 mg/m2 followed by paclitaxel 175 mg/m2 given over three hours [2]. The more active regimens are associated with enhanced cardiotoxicity; this toxicity can be avoided, however, by limiting the exposure to doxorubicin. The newer regimens have now been moved into phase III studies. Future progress for this disease will depend on the introduction of new agents. Two novel drugs are currently being investigated in randomised phase III trials as potentiators of Dox and/or paclitaxel. One is a monoclonal antibody from Genentech (Herceptin, trastuzumab) directed at the HER-2/neu oncogene, which is overexpressed in > 25% of breast cancers [3]. Recent results indicate that Herceptin in combination with paclitaxel (or with a Dox plus cyclophosphamide regimen) induces a higher response rate (RR) and prolongs the time to disease progression when compared to chemotherapy alone. The second agent N,N-diethyl-2[4-(phenylmethyl)-phenoxy] ethanamine.HCl (DPPE, BMS-217380-01), when combined with Dox, was associated with a higher RR than previously observed with Dox alone [4]. A randomized trial of Dox versus Dox plus DPPE is ongoing. The possible mechanisms underlying chemo potentiation by these agents are discussed. As new anthracycline/taxane combinations establish themselves in earlier stages of the disease, the need for effective, non-cross resistant salvage regimens will emerge. PMID- 10676566 TI - Current issues in phase I trials: new study designs and informed consent procedures. PMID- 10676568 TI - Determinants of protogenetic interval in a west Mediterranean rural population: La Alpujarra (southeast Spain). AB - This paper analyzes the protogenetic interval determinants and the influence on family size. The data came from La Alpujarra (Southeast Spain), consisting on a sample of 847 families marrying through-out the first half of the present century. The marital fertility includes 85% deliveries, 5% premarital births and the remaining 10% premarital conceptions. The protogenetic interval was clearly associated with the reproductive success since family size was nearly one child greater for short intervals (= < 16 months). The protogenetic interval largely depends on the occurrence of miscarriages preceding the first liveborn delivery as well as on maternal age. Consanguineous couples show slightly shorter intervals. A temporal decrease of protogenetic intervals was observed. PMID- 10676567 TI - New approaches in cancer treatment. AB - Major advances in cellular biology, genetics, pharmacology and immunology in the past decade are beginning to be translated into progress in cancer treatment. This progress is manifested by new cytotoxic drugs which have recently entered clinical practice (taxanes, topoisomerase I inhibitors, gemcitabine, vinorelbine, new purines), as well as the efficacy of monoclonal antibody therapies against the CD-20 antigen of B-cell lymphomas and the Her2/neu oncogene in breast cancer. Several new drugs in development are targeted at reversal or prevention of the multidrug resistance mechanism caused by expression of the MDR1 gene (P glycoprotein). Tumour angiogenesis as a target is being studied in several early clinical trials. As with many other biological therapies, the evaluation of these compounds and their integration with standard therapies presents a major challenge to clinical investigators. The emerging field of genomics and gene expression micro-arrays will provide enormous information about the biology of cancers. This technology offers great opportunities for the discovery of new therapeutic targets, which should provide a basis for the design and evaluation of many new agents in the coming decade. PMID- 10676570 TI - Finger dermatoglyphics in the Corsican population (France). AB - Finger pattern types, pattern intensity indices and finger ridge counts in 110 individuals (54 males and 56 females) from Corte in the central area of Corsica (France) were investigated. The comparison of the Corsican qualitative and quantitative digital dermatoglyphics with those from other samples of Mediterranean and European countries show a clearcut difference between Corsicans and Continental Italian populations and a great affinity between Corsicans and Sardinians. These results are regarded as compatible with the interpretation of archaeological, historical and genetic evidence. PMID- 10676569 TI - Comparisons of fatness indicators in Budapest children. AB - The aim of this study was to estimate the fatness level of Budapest children and youth in different ways and to compare these estimations using a large representative sample. Eighteen body measurements were taken on 2606 healthy boys and 2471 healthy girls aged between 3 and 18 years. About 20% of this sample was measured by the Futrex 5000A near infrared (NIR) spectrophotometer to assess the body fat percent (data of 419 boys and 462 girls aged between 5 and 18 years were analysed). Triceps skinfold thickness (TSF), sum of triceps, medial calf, subscapular and suprailiac skinfold thicknesses (SFS), body fat percent estimated according to Slaughter et al. (%BF), BMI (calculated from height and weight) and body fat percent assessed by NIR-method (NIR%BF) were compared. chi 2 tests of independence show significant connections among the distributions ranged by the five fatness indicators. However, correlation coefficients and standard errors indicate that strong relationships are only among the assessments based on skinfold thicknesses (r = 0.92-0.97, SEE = 1.8-2.6%). BMI and NIR%BF assess body fatness differently compared to skinfold thicknesses: r-values are moderate and SEE-values are relatively large (r = 0.59-0.87, SEE = 1.9-4.7%). These findings can be seen in both the boys and the girls. NIR%BF comparing to %BF significantly overpredicts body fat percent in the boys and significantly underpredicts it in the girls. BMI, height and weight are not in significant correlation with NIR%BF in the boys but there are moderate correlations in the girls. Our suggestion is that more research is needed with the use of NIR-method in children and adolescents, and it is necessary to refine prediction equations taking into consideration very carefully sex sand age differences. PMID- 10676571 TI - Dermatoglyphic characteristics of a population from the central Rhodopes (south Bulgaria). AB - The finger and palmar prints of a total of 386 individuals (182 males and 204 females) at an age between 10 and 18 years from the region of the villages Petkovo and Banite, situated along the river Malka Arda in the Central Rhodopes, were studied. The following dermatoglyphic traits were analyzed: pattern intensity index (PII), the main line or Cummin's index (MLI), the frequencies of the proximal palmar triradius (t), the true hypothenar patterns (Hy) and the accessory interdigital triradii (AIT). The specific dermatoglyphic complexes after Heet were also determined for the population under study. A peculiarity of this population is the appearance of a clearly expressed Eastern Complex (EC), which accounts for 53.1% in the males and for 48.8% in the females. These values of EC and the calculated dermatoglyphic distances (DD) after Heet show a similarity between the examined population and a number of populations from the Volga region, Northern Caucasus, Middle Asia and Siberia. This similarity could be explained with the preservation of the genetic heritage of the proto Bulgarians, one of the three main components of the modern Bulgarians. PMID- 10676572 TI - Study on pottical type, palmar and plantar digital formulae, hand clasping, arm folding, handedness, leg folding and stride type in the Daur population, China. AB - The pottical type, palmar and plantar digital formulae, hand clasping, arm folding, handedness, leg folding and stride type have been investigated on a sample of 143 male and 160 female students of the Daur population of Molidawa Banner, Inner Mongolia. The results of this study are the following: 1. the frequency of the hyperextensive pottical type is 49.17%, the relative length of index over annularis 12.21%, right hand clasping 45.87%, right arm folding 49.50%, right handedness 94.39%, right leg folding 72.28% and right stride type 44.88%, 2. pottical type, hand clasping, handedness, leg folding and stride type do not show significant sex differences, 3. there are some relations between hand clasping and arm folding as well as between arm folding and stride type, 4. compared with other population groups, the Daur population shows a low frequency of right hand clasping, a moderate frequency of right arm folding and a low frequency of left handedness. PMID- 10676573 TI - Is there a need for blood substitutes in the new millennium and what should we expect in the way of safety and efficacy? PMID- 10676574 TI - Gene therapy for hemophilia. AB - Hemophilia A and B are X-linked genetic disorders caused by deficiency of the coagulation factors VIII and IX, respectively. Because of the health hazards and costs of current product replacement therapy, much effort is devoted to the development of gene therapy for these disorders. Approaches to gene therapy for the hemophilias include: ex vivo gene therapy in which cells from the intended recipients are explanted, genetically modified to secrete Factor VIII or IX, and reimplanted into the donor; in vivo gene therapy in which Factor VIII or IX encoding vectors are directly injected into the recipient; and non-autologous gene therapy in which universal cell lines engineered to secrete Factor VIII or IX are enclosed in immuno-protective devices before implantation into recipients. Research into these approaches is aided by the many murine and canine models available. While problems of achieving high and sustained levels of factor delivery, and issues related to efficacy, safety and cost are still to be resolved, progress in gene therapy for the hemophilias has been encouraging and is likely to reach human clinical trial in the foreseeable future. PMID- 10676575 TI - Second-generation perfluorocarbon emulsion blood substitutes. AB - A novel series of perfluorocarbon (PFC) emulsions, based on perfluorodecalin (C10F18) and stabilised with up to 2.5% (w/v) of lecithin have been produced for evaluation as injectable, temporary respiratory gas-carrying blood substitutes. Some formulations contained 1.0% (w/v) of perfluorodimorpholinopropane (C11F22N2O2) to retard droplet growth through molecular diffusion (Ostwald Ripening). Other emulsions contained novel, amphiphilic fluorinated surfactants, such as, for example, the monocarbamate, C8F17C2H4NHC(O)(CH2CH2O)2Me (designated compound P6), at 0.1% (w/v) to enhance stability. Emulsions were prepared by homogenisation, were steam sterilisable and were stable for > 300 days (25 degrees C). Injection of rats (7.5 ml kg-1 b.w.) with emulsions produced significant (P < 0.05), transient increases in liver and spleen weights. One emulsion inhibited phorbol 12-myristate 13-acetate (PMA)-stimulated, Luminol enhanced, chemiluminescence of human polymorphonuclear leucocytes (PMNL) in vitro, suggesting possible applications in ischaemic tissues for suppressing PMNL mediated inflammation. The P6 fluoro-surfactant inhibited spontaneous platelet aggregation in hirudin-anticoagulated human blood in vitro, suggesting possible applications as an anti-thrombotic agent. PMID- 10676576 TI - Sustained drug release characteristics of biodegradable composite poly(d,l)lactic acid-poly(l)lactic acid microcapsules containing ciprofloxacin. AB - Ciprofloxacin polylactic microcapsules were prepared by the phase separation process. Two types of polylactic acid, poly(d,l)lactic acid and poly(l)lactic acid were combined as membrane materials to prevent the aggregation which happened frequently in the phase separation process. The polymer compositions of the microcapsules can influence the release rate of Ciprofloxacin. The optimal release rate of the drug can be obtained by modifying microcapsule compositions. Poly(d,l)lactic acid is superior in slowing the rate of drug release than poly(l)lactic acid. However, poly(l)lactic acid is necessary in the preparation of the microcapsules to prevent aggregation. PMID- 10676577 TI - Preparation of biodegradable microspheres of testosterone with poly(D,L-lactide co-glycolide) and test of drug release in vitro. AB - Biodegradable microspheres formulation of testosterone (T) can be used as a new physiological approach for androgen replacement in hypogonadal men. In this study, poly(D,L-lactide-co-glycolide) (PLGA) microspheres containing T were prepared by a solvent-evaporation/solvent-diffusion process and the drug release tests of the microspheres were carried out in vitro. T/PLGA microspheres with good yield, desired size and satisfied drug loading were obtained. A significant testosterone sustained release was shown in the drug release tests in vitro. Since PLGA microspheres preparations are normally sterilized by colbat-60 irradiation, the effects of 25 kGy colbat-60 irradiation on physicochemical properties and in vitro drug release profile of T/PLGA microsphere were investigated. The results showed that the irradiation didn't have any effects on the physicochemical properties of T. Though about one-third decrease in molecular weight of PLGA was caused by the irradiation, no significant changes were observed on the drug release profile in vitro. PMID- 10676578 TI - Evaluation of heparin immobilized chitosan-PEG microbeads for charcoal encapsulation and endotoxin removal. AB - A technique is described to encapsulate activated charcoal for hemoperfusion to be used in an artificial liver support. Activated charcoal was encapsulated within chitosan-PEG matrix and subsequently surface modified with PGE1 or heparin (hep-AC-PEGCB) via the glutaraldehyde functionalities. This novel matrix was used as the supports for perfusion of endotoxin, under a flow rate of 30 ml/mt. Endotoxin adsorption was quantitatively measured by the method of Limulus Amebocyte lysate test. It seems, the hep-AC-PEGCB may be a good adsorbent system for the removal of toxic endotoxin, and the system may be useful for detoxification of blood. The hep-AC-PEGCB matrix had improved biocompatibility as demonstrated from their hemolytic potential and charcoal release. However, further studies are needed to determine their behaviour under clinical conditions. PMID- 10676579 TI - The anticalcification effect of polyethylene glycol-immobilized on hexamethylene diisocyanate treated pericardium. AB - Pathologic calcification is thought to be the main cause of failure in the present generation tissue valves fabricated from glutaraldehyde pretreated bovine pericardium (BP). The present investigation describes the in vitro calcification and enzymatic degradation of bovine pericardia after hexamethylene diisocyanate (HMDIC) crosslinking and subsequent modification with polyethylene glycol. The enzymatic degradation of these treated surfaces were monitored by scanning electron micrography and tensile strength measurements. Various proteases, such as alpha-chymotrypsin, bromelain, esterase, trypsin and collagenase were investigated for tissue stability. Incubation of these enzymes with crosslinked pericardia had variably reduced their tensile strength. Among these treated surfaces, polyethylene glycol (PEG) grafted BP via isocyanate functionalities had retained maximum strength. The PEG modified tissues had also indicated a substantial reduction in calcification, when compared to other treated tissues. Further, the biocompatibility of various pericardial tissues were established by platelet adhesion and octane contact angle measurements. It is assumed that the PEG modification of pericardium may interfere with the cellular activation of injury (platelets) to reduce tissue associated calcification. In conclusion, it seems the PEG modification of bovine pericardium via HMDIC may provide new ways of controlling tissue biodegradation and calcification. However, more in vivo studies are needed to develop applications. PMID- 10676580 TI - Polycation-coated polyanion microspheres of urease for urea hydrolysis. AB - Urease (EC 3.5.1.5) was immobilized within polyanionic carboxymethylcellulose/alginate (CMC/Alg) microspheres coated with a cationic polysaccharide, chitosan (C). Coating with chitosan improved the mechanically durability of the polyanionic microspheres, as well as increased enzyme immobilization yield [approximately 0.4 mg.mL-1 gel]. The effects of chitosan coating and CMC/Alg ratio on the water uptake and spherical morphology of the microspheres were investigated. The optimal pH of urease was not extensively affected by the immobilization procedure. However, the optimal temperature of urease activity increased upto 60 and 65 degrees C within CMC/Alg and C(CMC/Alg) microspheres, respectively, while the optimum for the free enzyme was 50 degrees C. The half life (t1/2) and deactivation rate constant (kd) of free urease were 79 min and 8.77 x 10(-3) min-1, respectively, whilst the t1/2 and kd values of urease within polyanion and polycation-coated polyanion microspheres were 142 min and 4.88 x 10(-3).min-1, and 179 min and 3.87 x 10(-3).min-1, at 80 degrees C, respectively. While the activation energy of the hydrolysis reaction of free urease was found to be 11.86 kJ.M-1.dm-3, it increased to 18.91 and 20.02 kJ.M 1.dm-3, for the immobilized urease within CMC/Alg and C(CMC/Alg) microspheres, respectively. The free enzyme exhibited K(m) and Vmax values of 2.85 mM.dm-3 and 31.9 mM.dm-3.s-1.g-1p-1, respectively, whilst the K(m) and Vmax for urease within polyanion and polycation-coated polyanion microspheres were 3.94 mM.dm-3 and 73.4 mM.dm-3.s-1.g-1.p-1, and 4.22 mM.dm-3 and 81.4 mM.dm-3.s-1.g-1.p-1, in the same order. C(CMC/Alg) microspheres showed a nearly stable urease activity of around 80-85% of the initial maximum activity, after the first 100 minutes. PMID- 10676581 TI - Pain perception and response: central nervous system mechanisms. AB - Although several decades of studies have detailed peripheral and ascending nociceptive pathways to the thalamus and cerebral cortex, pain is a symptom that has remained difficult to characterize anatomically and physiologically. Positron emission tomography (PET) and functional magnetic imaging (fMRI) have recently demonstrated a number of cerebral and brain stem loci responding to cutaneous noxious stimuli. However, intersubject variability, both in the frequency and increased or decreased intensity of the responses, has caused uncertainty as to their significance. Nevertheless, the large number of available imaging studies have shown that many areas with recognized functions are frequently affected by painful stimuli. With this evidence and recent developments in tracing central nervous system connections between areas responding to noxious stimuli, it is possible to identify nociceptive pathways that are within, or contribute to, afferent spino-thalamo-cortical sensory and efferent skeletomotor and autonomic motor systems. In this study it is proposed that cortical and nuclear mechanisms for pain perception and response are hierarchically arranged with the prefrontal cortex at its highest level. Nevertheless, all components make particular contributions without which certain nociceptive failures can occur, as in pathological pain arising in some cases of nervous system injury. PMID- 10676582 TI - Predicting dementia in the elderly: a physician-friendly formula. PMID- 10676583 TI - Predicting who will develop dementia in a cohort of Canadian seniors. AB - OBJECTIVES: We examined whether easily attainable variables were useful in predicting who became demented over a five year period and determined the rates of incident dementia for different categories of mild cognitive impairment. METHODS: This was a cohort study of subjects recruited nationally in a population based survey of Canadians 65 years and older (the Canadian Study of Health and Aging). After standardized clinical assessments, a subset of subjects (n = 1782) was categorized as not demented at time one. Identical study methods allowed a reassessment of the cognitive status of surviving subjects (n = 892) five years later. RESULTS: Three baseline variables (Modified Mini Mental State (3MS) score, subject's age, and an informant's report of the presence of memory problems) were statistically significant predictors of the development of a dementia. An equation incorporating these three variables had a sensitivity of 79% and a specificity of 56% for predicting dementia among survivors at time two. An equation substituting the MMSE for the 3MS showed similar results. The various categories of mild cognitive impairment examined showed significantly different likelihoods for the subsequent development of a dementia. Some categories with a higher dementia risk were characterized by inclusion criteria requiring neuropsychological test scores that were greater than one standard deviation (SD) below the mean of age based normative data. CONCLUSION: In the absence of extensive laboratory, radiologic or neuropsychological tests, simple variables that can be easily determined in the course of a single clinical encounter were useful in predicting subjects with a higher risk of developing dementia. Attempts to use neuropsychological results to predict the development of dementia should look for significant impairments on age-standardized tests. PMID- 10676584 TI - Use of ambulatory electrocardiography for the detection of paroxysmal atrial fibrillation in patients with stroke. Canadian Task Force on Preventive Health Care. AB - BACKGROUND: Patients with stroke commonly undergo investigations to determine the underlying cause of stroke. These investigations often include ambulatory electrocardiography to detect paroxysmal atrial fibrillation. There is conflicting evidence in the literature regarding whether routine ambulatory electrocardiography should be performed in all or selected stroke patients. This paper reviews the available evidence on (1) the yield of ambulatory electrocardiography in detecting paroxysmal atrial fibrillation in patients with stroke or transient ischemic attack and (2) the effectiveness of anticoagulation in preventing recurrent stroke in patients with paroxysmal atrial fibrillation. METHODS: A MEDLINE search for primary articles was performed, and the references were reviewed manually. In addition, citations were obtained from experts. The evidence was systematically reviewed using the evidence-based methodology of the Canadian Task Force on Preventive Health Care. RESULTS: Ambulatory electrocardiography can detect atrial fibrillation not found on initial electrocardiogram in between 1% and 5% of people with stroke. Ambulatory electrocardiography is generally safe. The risk of recurrent stroke in the setting of paroxysmal atrial fibrillation is uncertain, but appears to be similar to that seen with chronic atrial fibrillation (about 12% per year). Therapy with warfarin may reduce this risk by about two-thirds as compared to placebo. The annual risk of major bleeding with warfarin therapy is between 1% and 3% but rates for individual patients depend on various specific risk factors. INTERPRETATION: There is insufficient evidence to recommend for or against the use of ambulatory electrocardiography for the detection of paroxysmal atrial fibrillation in either selected or unselected patients with stroke (C Recommendation). There is fair evidence to recommend therapy with warfarin for patients with stroke and paroxysmal atrial fibrillation (B Recommendation). PMID- 10676585 TI - Predictors of poor outcome in patients with a spontaneous cerebellar hematoma. AB - BACKGROUND AND PURPOSE: The authors studied the clinical and neuroimaging features of cerebellar hematomas to predict poor outcome using comprehensive statistical models. METHODS: We retrospectively reviewed clinical and neuroimaging features in 94 patients with spontaneous cerebellar hematomas to identify predictive features for a poor neurologic outcome, defined as death or dismissal to long-term care facility. Data were analyzed using chi square and Fisher's exact test with calculation of odd's ratios together with 95% confidence intervals. RESULTS: Clinical and neuroradiologic predictors for a poor outcome at p < 0.05 were admission systolic blood pressure > 200 mm Hg, hematoma size > 3 cm, visible brain stem distortion, and acute hydrocephalus. Presenting findings predicting subsequent death at p < 0.05 were abnormal corneal and oculocephalic responses, Glasgow coma sum score less than 8, motor response less than localization to pain, acute hydrocephalus and intraventricular hemorrhage. CONCLUSION: A tree-based analysis model using binary recursive partitioning showed that cornea reflex, hydrocephalus, doll's eyes, age, and size were the most important discriminating factors. Absent corneal reflexes on admission highly predicts poor outcome (86 percent, confidence limits 67-96 percent). When a cornea reflex is present, acute hydrocephalus predicts poor outcome but only when doll's eyes are additionally absent. PMID- 10676586 TI - Generic substitution for brand name antiepileptic drugs: a survey. AB - BACKGROUND/OBJECTIVE: There are presently 26 different generic preparations for five brand name antiepileptic drugs (AEDs) on the Canadian market with others likely to be released in the near future. The purpose of this review is to examine the basis for the controversy surrounding generic substitution for brand name antiepileptic drugs, to present the results of a survey of neurologists' and patients' attitudes toward generic substitution and to increase neurologists' awareness of the issues. METHODS: The current federal and provincial regulations pertaining to generic drug approval and substitution are reviewed. Published anecdotal and survey reports of the effectiveness and tolerability of generic substitution for AEDs are reviewed. A pilot questionnaire survey of 83 patients from four adult epilepsy clinics and 46 neurologists from across Canada was undertaken to determine attitudes toward generic substitution. RESULTS AND CONCLUSIONS: Several authors have suggested that some AEDs, particularly those with a narrow therapeutic index, may pose problems with generic substitution. Although generic AEDs are lower in price, possible increased side effects and morbidity and the need for closer monitoring could partially offset the cost savings. The results of our survey highlight significant unawareness of the process of generic substitution among both patients and neurologists and reveal a general level of discomfort among neurologists to prescribe generic AEDs. Further data should be obtained about the potential consequences of generic substitution in epilepsy patients. PMID- 10676587 TI - Craniotomy revisited: techniques for improved access and reconstruction. AB - OBJECTIVE: To describe simple modifications of the technique of opening and closure of the craniotomy to improve basal exposure and reconstruction. METHODS: The modifications involve: a) additional soft-tissue dissection which is carried downward to the base of the ear and to the orbital rim, exposing the orbital rim and malar eminence without removing the bone; b) cutting the bone flap so that 'bridges' of bone remain that help to stabilize the flap when it is returned to the cranium at the end of the operation; c) the wedging of bone chips between the bone flap and native cranium at the time the bone is being reaffixed so as to provide firm stability by diminishing movement of the bone flap; d) the use of bone dust and bone chips mixed with the patient's blood to seal and bridge the gap between the bone flap and the native bone; e) reattachment of the temporalis muscle with the bone flap sutures. An 'inlay' technique of duraplasty is also described. RESULTS AND CONCLUSION: These simple modifications of craniotomy provide better basal exposure and reconstruction with little additional operating time at no additional cost. PMID- 10676588 TI - Postictal aphasia and paresis: a clinical and intracerebral EEG study. AB - BACKGROUND: We examined the lateralizing value of postictal language and motor deficits and studied their underlying mechanisms. PATIENTS AND METHODS: The total sample consisted of 35 patients (26 temporals, 8 frontals, 1 parietal) with a good postsurgical outcome (Engel's class I and II). Postictal examination was blindly reviewed on videotapes. In 15 cases (29 seizures), postictal language manifestations were analyzed in relation with the diffusion of the epileptic discharge recorded by intracerebral EEG. Language dominance was determined by the intracarotid amobarbital test. RESULTS: Postictal aphasia was observed only when (1) seizure originated in the dominant hemisphere and (2) ictal activity spread to language areas (Wernicke and/or Broca areas). When the epileptic focus was in the nondominant hemisphere, no postictal aphasia was observed even if there was secondary generalization of ictal activity affecting the language areas of the dominant hemisphere. Postictal motor deficits also had a strong lateralizing value even when seizures were secondarily generalized. CONCLUSION: Postictal aphasia in temporal epilepsies and postical motor deficits in temporal and extra temporal epilepsies provided excellent lateralizing information. Postictal deficits appear to be the result of inhibitory mechanisms induced by previous ictal activity of the structures related to these functions. PMID- 10676589 TI - Post-traumatic cervical dystonia: a distinct entity? AB - BACKGROUND/OBJECTIVE: The incidence of head/neck trauma preceding cervical dystonia (CD) has been reported to be 5-21%. There are few reports comparing the clinical characteristics of patients with and without a history of injury. Our aim was to compare the clinical characteristics of idiopathic CD (CD-I) to those with onset precipitated by trauma (CD-T). METHODS: We evaluated 114 consecutive patients with CD over a 9-month period. All patients were interviewed using a detailed questionnaire and had a neurological examination. Their clinical charts were also reviewed. RESULTS: Fourteen patients (12%) had mild head/neck injury within a year preceding the onset of CD. Between the two groups (CD-I and CD-T), the gender distribution (F:M of 3:2), family history of movement disorders (32% vs. 29%), the prevalence of gestes antagonistes (65% vs. 64%), and response to botulinum toxin were similar. There were non-specific trends, including an earlier age of onset (mean ages 43.3 vs. 37.6), higher prevalence of neck pain (86% vs. 100%), head tremor (67% vs. 79%), and dystonia in other body parts (23% vs. 36%) in CD-T. CONCLUSIONS: CD-I and CD-T are clinically similar. Trauma may be a triggering factor in CD but this was only supported by non-significant trends in its earlier age of onset. PMID- 10676590 TI - Difference of disability between electrophysiologic subgroups of essential tremor. AB - OBJECTIVE: The aim of the study was to test the validity of the controversial subdivision of essential tremor (ET) patients into electrophysiological subgroups. METHODS: We evaluated a hundred patients with ET using surface electromyographic (EMG) recordings of antagonists forearm muscles and distinguished three groups: the first group showed synchronous activity of antagonistic muscles, the second showed alternating activity of antagonist muscles; and the third group consisted of patients whose EMG recordings were not compatible with the other two groups. We compared patients with synchronous and alternating activity in terms of sex, age at onset, duration of illness, family history of tremor, symmetry and frequency of tremor, and the scores of a disability scale. RESULTS: The only significant difference between the patients with synchronous and alternating activity was that the patients with synchronous activity were more disabled. CONCLUSION: This result adds to the evidence for distinct electrophysiological subgroups of ET with distinct clinical properties. PMID- 10676591 TI - Clinical and electromyographic examinations of patients with essential tremor. AB - BACKGROUND: It is believed that no clinical differences exist among essential, familial and senile tremor, or between the tremor with synchronous or alternating electromyographic activity. The aim of this study was to evaluate the clinical and electromyographic findings in a large group of patients with different types of essential tremor. METHODS: Two hundred and twenty patients with sporadic, familial or senile variants of essential tremor were examined. According to the electromyographic activity recorded from the antagonistic muscles, the patients were subdivided into a group with synchronous (SYN) and a group with alternating (ALT) activity. The historical aspects of the disease were noted, and a detailed neurological examination was performed. RESULTS: A widespread tremor involving upper and lower limbs and 3-4 different anatomical regions was typical for familial tremor. It also had higher amplitude than the sporadic and senile tremor. ALT tremor had a higher amplitude and longer burst duration than SYN and more often involved lower limbs. Rest tremor was common in the ALT group. Overall, ALT tremor was more common than previously supposed. CONCLUSION: The familial and ALT tremors are more disabling than other types of essential tremor. Since electromyographic ALT activity is common in essential tremor, its presence does not reliably distinguish essential and Parkinsonian tremor. PMID- 10676592 TI - Traumatic carotid-cavernous fistula. PMID- 10676593 TI - Hemorrhagic moyamoya disease during pregnancy. AB - BACKGROUND: Intracranial hemorrhage in pregnant patients with Moyamoya disease is rare. We review the case of one such patient who presented with pre-eclampsia and a catastrophic intracerebral hemorrhage in order to highlight the associated management difficulties. METHODS: A case of a pregnant (31 weeks) female brought to the emergency department with hypertension and a progressive decrease in her level of consciousness is presented. She rapidly developed a dilated right pupil and left extensor posturing. A CT scan of her head showed a large putamenal intracerebral hemorrhage. She was intubated, ventilated and given intravenous mannitol and magnesium sulfate. She underwent a simultaneous craniotomy and Cesarean section. Post-operatively the patient's ICP and jugular venous saturation were monitored in the intensive care unit. RESULTS: The patient delivered a 1185 g infant who did well. The patient's ICP was well controlled until the tenth post-operative day when she developed malignant brain edema and died. CONCLUSION: This case highlights three important points. First, simultaneous craniotomy and Cesarean section can be performed. Second, intraoperative control of bleeding Moyamoya vessels is described. Third, the difficult post-operative management of these cases is highlighted. The literature regarding Moyamoya disease and pregnancy is reviewed and some recommendations for the management of this rare but potentially deadly condition are presented. PMID- 10676594 TI - Ogilvie's syndrome as a rare complication of lumbar disc surgery. AB - BACKGROUND: In this study we report a rare complication after lumbar surgery, Ogilvie's syndrome, that presents as acute colonic dilatation in the absence of mechanical obstruction. CASE: A 43-year-old obese woman underwent lumbar surgery for L4-L5 lumbar disc herniation. The patient complained of persistent abdominal distention and lack of bowel sounds. Plain radiography and ultrasonography revealed massive dilatation of the colon. Nasogastric aspiration was initiated and all analgesic drugs were withdrawn. Abdominal distention gradually disappeared within three days. CONCLUSIONS: Only three cases of Ogilvie's syndrome following lumbar spinal surgery have been reported in the literature. In our case obesity, chronic constipation, and narcotic drugs were the most likely precipitating causes. Ogilvie's syndrome may resolve with conservative treatment, but if the cecal diameter continues to increase, colonoscopy or laparotomy may be needed to prevent perforation of colon. PMID- 10676595 TI - Neurology and neurosurgery at the Montreal General Hospital 1960-1980. PMID- 10676596 TI - Re: Management of Parkinson's disease: a review of current and new therapies. Tilak Mendis, Oksana Suchowersky, Anthony Lang, Serge Gauthier. Can J Neurol Sci 1999;26:89-103. PMID- 10676597 TI - Amiodarone: what have we learned from clinical trials? AB - Amiodarone is an antiarrhythmic agent commonly used in the treatment of supraventricular and ventricular tachyarrhythmias. This paper reviews clinical trials in which amiodarone was used in one of the treatment arms. Key post myocardial infarction trials include EMIAT and CAMIAT, both of which demonstrated that amiodarone reduced arrhythmic but not overall mortality. In patients with congestive heart failure (CHF), amiodarone was associated with a neutral survival in CHF/STAT and improvement in survival in GESICA. In patients with nonsustained ventricular tachycardia, the MADIT trial demonstrated that therapy with an implantable cardioverter-defibrillator (ICD) improved survival compared with the antiarrhythmic drug arm in such patients, most of whom were taking amiodarone. In sustained VT/VF patients, the CASCADE trial demonstrated that empiric amiodarone lowered arrhythmic recurrence rates compared with other drugs guided by serial Holter or electrophysiologic studies. Several trials including AVID, CIDS, and CASH have demonstrated the superiority of ICD therapy compared with empiric amiodarone in improving overall survival. Clinical implications of these trials are discussed. PMID- 10676598 TI - Five-year mortality in patients with acute chest pain in relation to smoking habits. AB - BACKGROUND: Smoking is one of the major risk indicators for development of coronary artery disease, and smokers develop acute myocardial infarction (AMI) approximately a decade earlier than nonsmokers. In smokers with established coronary artery disease, quitting smoking has been associated with a more favorable prognosis. However, most of these studies comprised younger patients, the majority of whom were males. HYPOTHESIS: The purpose of the study was to determine mortality, mode of death, and risk indicators of death in relation to smoking habits among consecutive patients admitted to the emergency department with acute chest pain. METHODS: In all, 4,553 patients admitted with acute chest pain to the emergency department at Sahlgrenska University Hospital during a period of 21 months were included in the analyses and were prospectively followed for 5 years. RESULTS: Of these patients, 36% admitted current smoking. They were younger and had a lower prevalence of previous cardiovascular diseases than did nonsmokers. The 5-year mortality was 19.4% among smokers and 24.9% among non smokers (p < 0.0001). However, when adjusting for difference in age, smoking was associated with an increased risk [relative risk (RR) 1.51; 95% confidence interval (CI) 1.32-1.74; p < 0.0001]. Among patients presenting originally with chest pain, the increased mortality for smokers was more pronounced in patients with non-acute than acute myocardial infarction (AMI). Among patients who died, death in smokers was less frequently associated with new-onset myocardial infarction (MI) and congestive heart failure. Among those who smoked at onset of symptoms and were alive 1 year later, 25% had stopped smoking. Patients with a confirmed AMI who continued smoking 1 year after onset of symptoms had a higher mortality (28.4%) during the subsequent 4 years than patients who stopped smoking (15.2%; p = 0.049). CONCLUSION: In consecutive patients admitted to the emergency department with acute chest pain, current smoking was significantly associated with an increased risk of death during 5 years of follow-up. Among patients who died, death in smokers was less frequently associated with new-onset MI and congestive heart failure than was death in nonsmokers. PMID- 10676599 TI - Intravenous Optison (FS069) enhances pulmonary vein flow velocity signals: a multicenter study. AB - BACKGROUND: Pulmonary vein spectral Doppler signals to characterize ventricular diastolic and systolic function, though often difficult to obtain, can be enhanced using contrast agents. HYPOTHESIS: The objective of this study was to determine the efficacy of the intravenous contrast agent Optison for enhancement of Doppler signals in patients with poor signals on two-dimensional echocardiographic examinations. METHODS: Enhancement of pulmonary venous flow was evaluated in 191 patients at 0.2, 0.5, 3.0, and 5.0 ml per injection. RESULTS: Greatest contrast enhancement for right and left pulmonary veins was observed at the highest doses. At 0.5 ml, conversion from inadequate to adequate was observed in right and left pulmonary veins in 48.0 and 79.3% of patients, respectively, while any degree of improvement was 54.4 and 65.8%, respectively. The adverse event rate (6.5%) was similar to a first-generation agent. CONCLUSION: The results demonstrate that Optison is a safe and effective contrast agent for improving visualization of pulmonary Doppler signals, especially the left pulmonary vein. PMID- 10676600 TI - Effects of early captopril therapy after myocardial infarction on the incidence of late potentials. AB - BACKGROUND: Late potentials (LP) on signal-averaged electrocardiography (SAECG), recorded 6 to 30 days after an acute myocardial infarction (AMI), identify patients at risk for late arrhythmic events. Angiotensin-converting enzyme (ACE) inhibitors have been shown to reduce ventricular remodeling and cardiovascular mortality after AMI. HYPOTHESIS: The aim of this study was to investigate the effect of early (< 24 h) administration of captopril on the presence of LP on Days 6-30 after AMI. METHODS: The study included 117 patients with a first AMI; 63 patients (53 men and 10 women, aged 59 +/- 12 years), 35 with an anterior and 28 with an inferior AMI (44 thrombolyzed), received early captopril therapy. The control group consisted of 54 age-matched patients (39 men and 15 women, aged 60 +/- 12 years), 19 with an anterior and 35 with an inferior AMI (31 thrombolyzed, p = NS), who did not receive early therapy with an ACE inhibitor. The mean left ventricular ejection fraction was similar in both groups (48 vs. 46%). Time domain analysis of SAECG was performed using a band-pass filter of 40-250 Hz. Late potentials were considered present if any two of three criteria were met: (1) Filtered QRS duration (QRSD) > 114 ms, (2) root-mean-square voltage of the last 40 ms of the QRS complex (RMS) < 20 microV, and (3) duration of low amplitude (< 40 microV) signal of the terminal portion of the QRS (LAS) > 38 ms. RESULTS: In the two groups of patients there were no differences in mean values of SAECG parameters. No patient was receiving any antiarrhythmic drugs. In the captopril group LPs were present in 9 of 63 patients (14%) and in the control group in 17 of 54 patients (31%) (p = 0.046). There was no difference in the number of patients with a patent infarct-related artery in the two groups (76 vs. 59%). CONCLUSION: Captopril treatment early after an AMI reduces the incidence of LPs recorded on Days 6-30 and may thus favorably affect the arrhythmogenic substrate. PMID- 10676601 TI - Cardiac autonomic tone and its relation to nonsustained ventricular tachyarrhythmias in idiopathic dilated cardiomyopathy. AB - BACKGROUND: In contrast to postinfarct patients, little is known about cardiac autonomic tone and its relation to spontaneous ventricular tachyarrhythmias in idiopathic dilated cardiomyopathy (IDC). Both heart rate variability (HRV) and baroreflex sensitivity (BRS) are indices of autonomic innervation of the heart. HYPOTHESIS: The aim of the present study was to determine the relation between cardiac autonomic tone assessed by HRV and BRS and spontaneous nonsustained ventricular tachycardia (NSVT) on Holter in a large patient population with IDC. METHODS: 24-h digital Holter recordings including HRV analysis and BRS testing were prospectively performed in 137 patients with IDC and preserved sinus rhythm. Mean age was 48 +/- 12 years, and mean left ventricular (LV) ejection fraction was 32 +/- 9%. The HRV analysis on Holter included the mean RR interval (RRm), the standard deviation of all normal RR intervals (SDNN), the square root of the mean of the squared differences between adjacent normal RR intervals (rMSSD), and the proportion of adjacent normal RR intervals differing more than 50 ms (pNN50). Testing for BRS was performed noninvasively using the phenylephrine method. RESULTS: Of 137 study patients, 42 (31%) had spontaneous NSVT on 24-h Holter. Compared with patients without NSVT, patients with NSVT on Holter had a higher New York Heart Association (NYHA) functional class (NYHA III: 40 vs. 18%, p < 0.01), a lower ejection fraction (29 +/- 9 vs. 34 +/- 9%, p = 0.01), and an increased LV end-diastolic diameter (69 +/- 8 mm vs. 66 +/- 7 mm, p = 0.03). The HRV variables rMSSD, pNN50, RRm, and BRS did not differ significantly between patients with and without spontaneous NSVT. Only SDNN on Holter was slightly lower in patients with versus without NSVT (106 +/- 45 vs. 121 +/- 46 ms, p = 0.08). CONCLUSIONS: Patients with IDC and spontaneous NSVT on Holter are characterized by a higher NYHA functional class, a lower LV ejection fraction, an increased LV end-diastolic diameter, and a tendency toward a lower SDNN value compared with patients without NSVT. The remaining measures of HRV including rMSSD and pNN50 reflecting primarily tonic vagal activity, as well as BRS reflecting predominantly reflex vagal activity, were similar in patients with and without NSVT. The prognostic significance of these findings in patients with IDC is currently under investigation in the Marburg Cardiomyopathy Study (MACAS) at our institution. PMID- 10676602 TI - The effect of amlodipine and enalapril on blood pressure and neurohumoral activation in hypertensive patients with Ribbing's disease (multiple epiphysal dystrophy). AB - BACKGROUND: In patients with Ribbing's disease (RD)--a form of multiple epiphysal dystrophy--hypertension is frequent, often severe, and accompanied by a relevant cardiac dysfunction. HYPOTHESIS: This study was undertaken to evaluate the contribution of the calcium antagonist amlodipine and of the angiotensin converting-enzyme inhibitor enalapril to blood pressure regulation by studying their effect on neurohormonal activation. METHODS: Fifty hypertensive patients with RD were studied. After a placebo run-in period of 4 to 6 weeks, patients were randomly assigned to receive either amlodipine (10 mg once daily) or enalapril (20 mg once daily) for 6 months. RESULTS: Both drugs significantly lowered blood pressure. Enalapril did not result in activation of the sympathetic system (as determined by measurement of the plasma norepinephrine level). On the other hand, the hypotensive effect of amlodipine occurred with an increase in heart rate and in the levels of plasma norepinephrine and angiotensin II. CONCLUSION: It is unclear whether amlodipine may reduce cardiac dysfunction in patients with RD. PMID- 10676603 TI - Comparison of left ventricular diastolic filling with myocyte bulk modulus using Doppler echocardiography and acoustic microscopy in pressure-overload left ventricular hypertrophy and cardiac amyloidosis. AB - BACKGROUND: The myocardial bulk modulus has been described as the constitutive properties of the left ventricular (LV) wall and is measured as rho V2 (rho = density, V = sound speed) using acoustic microscopy. HYPOTHESIS: The study was undertaken to assess the relationship between the myocyte bulk modulus and transmitral inflow patterns in patients with pressure-overload LV hypertrophy (LVH) and cardiac amyloidosis (AMD). METHODS: In 8 patients with LVH, 8 with AMD, and 10 controls without heart disease, the transmitral inflow pattern was recorded by Doppler echocardiography before death, and myocardial tissue specimens were obtained at autopsy. The tissue density and sound speed in the myocytes were measured by microgravimetry and acoustic microscopy, respectively. The diameters of the myocytes were measured on histopathologic specimens stained by the elastica Van Gieson method. RESULTS: In the subendocardium, the myocyte bulk modulus was larger in LVH (2.98 x 10(9) N/m2, p < 0.001) and smaller in AMD (2.61 x 10(9) N/m2, p < 0.001) than in the controls (2.87 x 10(9) N/m2). The myocyte diameter in LVH (26 +/- 1 microns) was larger than that in the control (21 +/- 1 microns, p < 0.001) and AMD (20 +/- 1 microns, p < 0.001). The bulk modulus in the subendocardial myocyte significantly correlated with the deceleration time (DT) of the early transmitral inflow (r = 0.689, p = 0.028 in control, r = 0.774, p = 0.024 in LVH, and r = 0.786, p = 0.021 in AMD). CONCLUSION: The changes in the myocyte elasticity as represented by the bulk modulus were limited to the subendocardial layers and may be related to relaxation abnormalities in LVH and a reduction in LV compliance in AMD. PMID- 10676604 TI - Malpositioned ventricular pacing lead in the left ventricle. PMID- 10676605 TI - Intercoronary communication between the circumflex and right coronary arteries. PMID- 10676606 TI - Refractory vasospasm with a malignant course. AB - We present a patient with two rare disorders, recurrent vasospastic angina leading to cardiac transplant and acute aortic occlusion. The patient had recurrent episodes of coronary vasospasm presenting with unstable angina, acute myocardial infarction, and sudden cardiac death in spite of adequate therapy with nitrates and calcium-channel blockers. He went on to have a cardiac transplant. The patient later presented with acute aortic occlusion with concomitant renal and mesenteric artery spasm. The circumstances of the presentation raise the possibility of a generalized vasospastic predisposition that is responsible for both events. Smoking, the only known major risk factor other than atherosclerosis, was noted to be temporally related to both events in our patient. PMID- 10676607 TI - Incomplete ventricular septal rupture following blunt chest trauma. AB - Nonpenetrating cardiac trauma should be considered in the diagnosis of electrocardiographic changes after road traffic accidents. Transesophageal echocardiography is the most useful noninvasive technique for the diagnosis of cardiac trauma. This paper reports the case of a patient with traumatic contusion of the ventricular septum following a fall from a 20 m height onto the roof of a car. PMID- 10676608 TI - Franz M. Groedel. PMID- 10676609 TI - Project GRACE (Guidelines for Resuscitation and Care at End-of-life). PMID- 10676610 TI - Medical futility. PMID- 10676611 TI - Advance Care Planning. Florida Legislature. PMID- 10676612 TI - Palliative care. PMID- 10676613 TI - Summary of Task Forces I, II, and III, and report of Task Force IV Education & Implementation. PMID- 10676614 TI - Magnetization transfer imaging of traumatic brain injury. AB - Magnetization transfer imaging (MTI) has been shown to be sensitive for the detection of white matter abnormalities in entities such as multiple sclerosis, progressive multifocal leukoencephalopathy, and wallerian degeneration. Our hypothesis was that MTI would detect traumatic white matter abnormalities (TWMA) and provide information additional to that obtainable with routine spin- and gradient-echo imaging. We hypothesized that the presence of TWMA defined by MTI would correlate with outcome following TBI. Twenty-eight victims of head trauma and 15 normal controls underwent magnetic resonance imaging including MTI. Magnetization transfer ratios (MTR) were calculated for areas of shearing injury and for normal-appearing white matter (NAWM) in locations frequently subject to diffuse axonal injury. Abnormal MTRs were detected in NAWM in eight patients. All eight had persistent neurologic deficits, including cognitive deficits, aphasia, and extremity weakness. Seven of the 28 patients had no abnormal findings on neurologic exam at discharge, transfer, or follow-up. None of these patients had an abnormal MTR in NAWM. In the remaining 13 patients, who had persistent neurologic deficits, no regions of abnormal MTR were detected in NAWM. MTI is a sensitive method for the detection of TWMA. Detection of abnormal MTR in NAWM that is prone to axonal injury may predict a poor patient outcome. The presence of normal MTR in NAWM in these areas does not necessarily confer a good outcome, however. PMID- 10676615 TI - Proton MR spectroscopy in children with acute brain injury: comparison of short and long echo time acquisitions. AB - The aim of this study was to evaluate comparatively the information given by proton magnetic resonance spectroscopy (MRS) with short echo time (TE 20 msec) stimulated echo acquisition mode and long TE (270 msec) point-resolved spectroscopy in predicting long-term outcome in children suffering from acute brain injury. At 1.5 T, we performed single-voxel proton MRS with both methods in occipital gray matter of 70 children. A linear discriminant analysis used to predict outcomes based on MRS variables was compared with actual neurologic outcome assigned at least 6 months after injury by a pediatric neurologist. Using peak area metabolite ratios and lactate presence, the short and long TE methods were equally predictive in children over 1 month of age. In neonates less than 1 month of age, the long TE method produced a higher percentage of correct outcome predictions (91%) than the short TE method (79%). The long TE method detected lactate more often in all age groups. PMID- 10676616 TI - Evaluation of intrathecal gadolinium-enhanced MR cisternography in a rabbit model of traumatic nasoethmoidal CSF fistula. AB - This pilot study details the feasibility of intrathecal gadopentetate dimeglumine (Gd) administration in the detection of posttraumatic cerebrospinal fluid (CSF) fistula in an animal model. Five rabbits were used in this study. An attempt was made to create a traumatic CSF fistula surgically via a nasal approach. Seven days following the procedure, images of the cranium in sagittal and coronal planes were obtained utilizing a 1.9 T magnetic resonance (MR) imaging scanner before and after intrathecal injection of 16 pmol Gd. Following the imaging study, the animals were euthanized and grossly sectioned coronally to search for fistula formation. One animal died on the third day following the surgical procedure. The other four rabbits underwent the MR and gross pathologic study. Diagnosis of the fistula by intrathecal Gd-enhanced MR imaging was successful in two rabbits; this finding was confirmed by gross pathologic examination. No fistula was detected on either intrathecal Gd-enhanced MR imaging or on pathologic study in the remaining two rabbits. Intrathecal enhanced MR cisternography is a potentially promising technique for the evaluation of posttraumatic CSF fistulae. PMID- 10676617 TI - Proton magnetic resonance spectroscopy of temporal lobe white matter in patients with histologically proven hippocampal sclerosis. AB - The purpose of this study was to assess temporal lobe white matter changes accompanying hippocampal sclerosis on magnetic resonance (MR) imaging using single-voxel 1H MR spectroscopy and to strengthen the hypothesis that these white matter changes are caused by myelin alterations. In 11 patients with histologically proven hippocampal sclerosis, preoperative coronal fluid attenuated inversion recovery images were visually assessed by two experienced neuroradiologists for hippocampal signal increase and size decrease, atrophy of collateral white matter, and temporal lobe gray/white matter demarcation loss. Single-voxel 1H MR spectroscopy of the white matter of each anterior temporal lobe was also performed, excluding the amygdala and hippocampus. The N-acetyl aspartate (NAA)/choline and NAA/creatine ratios were calculated. In 12 healthy volunteers both temporal lobes were spectroscopically examined. In all patients the excised hippocampi were histologically assessed for the presence of sclerosis, and the excised neocortical temporal lobes were examined for gray and white matter abnormalities. MRI abnormalities were found on the right in six patients, on the left in four, and one scan was normal. Hippocampal signal increase was seen in nine patients, hippocampal size decrease in ten, atrophy of collateral white matter in nine, and gray/white matter demarcation loss in six. A significant decrease in the NAA/choline ratio was found in temporal lobe white matter ipsilateral to the pathologic hippocampus (symptomatic side), compared with the contralateral, asymptomatic side (P < 0.01), and also compared with controls (P < 0.001). The ipsilateral NAA/creatine ratio was also significantly decreased (P < 0.05) compared with the contralateral side and the control subjects (P < 0.001). Histological examination showed hippocampal sclerosis to a different degree in all patients. Neither gliosis nor cortical dysplasia was found in the ipsilateral, symptomatic temporal lobe. Significant decrease in the mean of NAA/choline ratios is found in temporal lobe white matter of patients with histologically confirmed hippocampal sclerosis. As this indicates neuronal loss or dysfunction, the number of axons may be reduced, with associated decrease in myelin density. PMID- 10676618 TI - MRA of the abdominal aorta and lower extremities. AB - Atherosclerotic involvement of the aorta and lower extremity vessels is a common clinical problem, especially in developed countries. While x-ray angiography has been the method of choice for preoperative evaluation of patients with atherosclerotic disease, magnetic resonance angiography (MRA) is emerging as a powerful noninvasive tool that is capable of providing information critical to the care of these patients. The objective of this manuscript is to review the current state-of-the-art of MRA of the abdominal aorta and lower extremity vessels. The techniques are described, the clinical indications for MRA are discussed, and the diagnostic accuracy and pitfalls of the various methods are presented. PMID- 10676619 TI - Budd-Chiari syndrome: spectrum of appearances of acute, subacute, and chronic disease with magnetic resonance imaging. AB - The purpose of this study was to describe our collective experience in the magnetic resonance (MR) investigation of patients with proven acute, subacute, and chronic Budd-Chiari syndrome and to demonstrate the spectrum of appearances on T1- and T2-weighted as well as dynamic post-gadolinium spoiled gradient-echo imaging. All patients with proven Budd-Chiari syndrome who underwent MR examinations between June, 1992 and October, 1998 were included in the study. Fourteen patients were included in the study: four with acute, three with subacute, three with chronic, and four with acute superimposed on either subacute (two) or chronic (two) Budd-Chiari syndrome. MR imaging features were retrospectively evaluated to determine: a) liver morphology, b) pattern of signal intensity (SI) on T1-weighted images, c) pattern of SI on T2 weighted images, d) dynamic enhancement characteristics, e) presence or absence of visible venous thrombosis, and f) presence or absence of venous macroscopic collaterals. The MR findings were correlated with surgical, histopathological, and laboratory data to determine imaging characteristics related to the chronicity of the disease process. Hepatic venous thrombosis or absence of hepatic venous flow was demonstrated in all patients in the study. In the four patients with acute Budd Chiari syndrome, the liver periphery was moderately low signal on T1 and moderately high signal on T2-weighted images relative to the central liver; both early and late gadolinium-enhanced images revealed diminished peripheral enhancement. In the three patients with subacute Budd-Chiari syndrome, the liver periphery was moderately low signal on T1, and moderately high signal on T2 weighted images, while early and late gadolinium-enhanced images revealed heterogenously increased enhancement within the liver periphery. In the three patients with chronic Budd-Chiari syndrome, the SI differences between peripheral and central liver were minimal on T1- and T2-weighted images, and enhancement differences were also minimal. Extensive bridging intrahepatic and capsular venous collaterals were visualized in chronic cases. In the four patients with acute Budd-Chiari syndrome superimposed on more chronic disease, a combination of gadolinium enhancement patterns was observed on MR images. Enhancement patterns between central and peripheral liver were different for acute, subacute, and chronic Budd-Chiari syndromes, suggesting differentiation between these phases of the disease process. Application of this pattern approach permitted recognition of acute changes superimposed on more chronic disease. PMID- 10676620 TI - Polyposis syndromes of the gastrointestinal tract: MR findings. AB - We describe the magnetic resonance (MR) findings in patients with gastrointestinal polyposis syndromes using breath-hold T1-weighted sequences, both standard and with fat suppression, prior to and following gadolinium administration, and breathing-independent single-shot half-Fourier RARE T2 weighted sequences. Six patients with gastrointestinal polyposis syndromes underwent MR examination to investigate for the presence of metastatic disease. The appearances of the gastrointestinal polyps on noncontrast T1-weighted spoiled gradient-echo (SGE), T2-weighted (half-Fourier RARE) images, and early and late gadolinium-enhanced SGE images were determined. Other gastrointestinal findings and extragastrointestinal disease were also evaluated. Patients with the following gastrointestinal polyposis syndromes were included: familial polyposis (n = 3), Peutz-Jeghers syndrome (n = 1), Gardner's syndrome (n = 1), and neurofibromatosis (n = 1). Polypoid lesions in all patients exhibited signal intensity comparable to bowel on noncontrast images and enhanced similar to bowel on early and late gadolinium-enhanced images. Polyps larger than 2 cm, observed in one patient with familial polyposis and the patient with Gardner's disease, showed mild heterogeneity on late gadolinium-enhanced fat-suppressed images. Multiple colonic polyps ranging from 5 mm to 3 cm in diameter were observed in patients with familial adenomatous polyposis. A solitary 1.5 cm polyp associated with entero-enteric intussusception was observed in the patient with Peutz Jeghers syndrome. Gastric polyps ranging from 5 mm to 6 cm were observed in the stomach of the patient with Gardner's syndrome. Duodenal and jejunal neurofibromas ranging from 1 to 2 cm in diameter were present in the patient with neurofibromatosis. Extra gastrointestinal findings included an adrenal adenoma (1 patient), a pheochromocytoma (1 patient), and liver metastases (2 patients). Gastrointestinal polyps in patients with polyposis syndromes may be visualized on MR images employing breath-hold T1-weighted and breathing-independent snapshot T2 weighted techniques. Appreciation of polyp enhancement on post-gadolinium images is an important finding, which should help distinguish polyps from bowel contents. PMID- 10676621 TI - Comparison of short inversion time inversion recovery (STIR) and fat-saturated (chemsat) techniques for background fat intensity suppression in cervical and thoracic MR imaging. AB - The purpose of this study was to compare short inversion time inversion recovery (STIR) fast spin-echo (FSE), and fat-saturated T2-weighted FSE sequences in terms of uniformity of fat suppression and lesion conspicuity for magnetic resonance (MR) imaging of the neck and thorax. STIR FSE and fat-saturated T2-weighted FSE images were scored for uniformity of fat suppression (n = 40) and lesion conspicuity (n = 35). Five-point rank score analyses were utilized by three experienced radiologists. The mean scores of STIR and fat-saturated FSE techniques for uniformity of fat suppression were 4.3 and 2.3, respectively (P < 0.0001). The mean scores of STIR and fat-saturated FSE techniques for lesion conspicuity were 4.2 and 3.5, respectively (P < 0.0001). Insufficient fat suppression was prominent in the mandible, supraclavicular region, anterior mediastinum, epipericardial fat, and subdiaphragmatic fat. In addition, fat saturated T2-weighted FSE showed inadvertent water suppression in 25%. The STIR FSE technique was superior to the fat-saturated FSE technique for cervical and thoracic MR imaging. PMID- 10676622 TI - Statistical analysis of multi-subject fMRI data: assessment of focal activations. AB - A simple procedure for analyzing multi-subject functional magnetic resonance imaging (fMRI) data is proposed. In the first step, a voxel-wise t-test across standardized z-maps is performed to identify areas that are consistently activated across subjects. In the second step, for each area, individual mean z scores are calculated and subsequently subjected to an analysis of variance. An example is provided. PMID- 10676623 TI - 3D gadolinium-enhanced MRA: evaluation of hepatic vasculature in children with hepatoblastoma. AB - We used contrast-enhanced three-dimensional magnetic resonance angiography (3D MRA) modified for pediatric use to evaluate the hepatic vasculature prior to partial hepatectomy in five consecutive children with hepatoblastoma. Modifications included non-breath-hold technique in four of the five children who were sedated. The single breath-hold technique was performed in only one awake child. Scan delay times were based on contrast infusion time rather than total infusion time. The hepatic artery, portal vein, and inferior vena cava were identified in all patients. MRA findings were confirmed by conventional angiography in one patient and by surgery in all. Contrast-enhanced 3D MRA is a useful and rapid technique prior to partial hepatectomy in patients with hepatoblastoma. PMID- 10676624 TI - Comparison of artifacts produced from carbon fiber and titanium alloy needles at 1.5 T MR imaging. AB - A novel coaxial carbon fiber-based biopsy needle set was investigated in phantom experiments and compared with a commercially available, magnetic resonance (MR) compatible titanium alloy set using MR imaging at 1.5 T. Image artifacts observed with different MR sequences were assessed. It was found that the carbon fibers produced distinctly smaller image artifacts compared with the titanium needle. Depending on the type of MR sequence, the relative range of artifact size ratios between the carbon and titanium needles was between 0.7 (spin-echo sequence) and 0.4 (gradient-echo sequence) with the needles oriented perpendicular to the main magnetic field. Carbon fiber composites are promising materials for the design and construction of MR-compatible instruments. PMID- 10676625 TI - Induction of apoptotic cell death and in vivo growth inhibition of human cancer cells by a saturated branched-chain fatty acid, 13-methyltetradecanoic acid. AB - A saturated branched-chain fatty acid, 13-methyltetradecanoic acid (13-MTD), was purified from a soy fermentation product, which was used by many cancer patients as a treatment supplement. Our preliminary study indicated that 13-MTD could induce cell death in human cancer cell lines K-562, MCF7, DU 145, NCI-SNU-1, SNU 423, NCI-H1688, BxPC3, and HCT 116. The ID50 dosage of 13-MTD for these tumor cells ranged from 10 to 25 microg/ml. Further investigation revealed that 13-MTD caused tumor cell death through rapid induction of apoptosis, which could be detected 2 h after the treatment of tumor cells with 13-MTD. Xenograft tumors of prostate carcinoma cell line DU 145 and hepatocarcinoma LCI-D35 were orthotopically implanted into nude mouse prostate and liver, respectively. 13-MTD was administered p.o. once daily to the implanted mice for approximately 40 days. Our results showed that 13-MTD could effectively inhibit the growth of orthotopic tumor implants of both cell lines compared with control groups. The average inhibition rate was 84.6% for DU 145 and 65.2% for LCI-D35 (P < 0.01). LD50 test results showed that mice could well sustain the oral feeding of 5 g/kg/day without observable anomaly. Our preliminary data demonstrated that 13-MTD could effectively inhibit in vitro and in vivo growth of various cancer cell lines by inducing apoptosis without significant toxic side effects, suggesting 13-MTD as a potential candidate for chemotherapy of human cancers. PMID- 10676626 TI - Expression of a highly conserved protein, p27BBP, during the progression of human colorectal cancer. AB - The highly conserved protein p27BBP is a cytoplasmic interactor of integrin beta4 expressed in epithelia. p27BBP is found in two pools: one nuclear pool enriched in the perinucleolar region, and one cytoplasmic pool. Deletion of p27BBP in yeast is lethal as a result of loss of the ribosomal 60S subunit. The aim of this study was to investigate the distribution of p27BBP in gut epithelium and its behavior during progression of human colorectal carcinomas. Results indicated that p27BBP is high in rapidly cycling cells and decreased in villous cells committed to apoptotic cell death. In dysplastic adenomas and carcinomas, p27BBP displayed a large increase of its nucleolar component that was superimposable to argyrophylic nucleolar organizing region-associated proteins and was associated with the nuclear matrix. Western blotting confirmed increased p27BBP in dysplastic adenomas and in carcinomas. In particular, p27BBP increased progressively from adenomas to carcinomas and, in the latter, was related to the tumor stage. The overexpression of p27BBP corresponded to mRNA up-regulation in carcinomas, supporting the idea of transcriptional or post-transcriptional regulation of its expression. Results suggested that p27BBP alterations are an early event in the transition from benign to malignant colorectal phenotypes and provide a novel tool in surgical pathology. PMID- 10676627 TI - p53-dependent global genomic repair of benzo[a]pyrene-7,8-diol-9,10-epoxide adducts in human cells. AB - The global genomic repair of DNA adducts formed by the human carcinogen (+/-) anti-benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE) has been studied by 32P postlabeling in human fibroblasts in which p53 expression can be regulated. At low BPDE adduct levels (10-50 adducts/10(8) nucleotides), repair was rapid and essentially complete within 24 h in p53+ cells, whereas no repair was detected within 72 h in similarly treated p53- cells. At 10-fold higher BPDE adduct levels, repair under both conditions was rapid up to 8 h, after which a low level of adducts persisted only in p53- cells. These results demonstrate a dependence on p53 for the efficient repair of BPDE adducts at levels that are relevant to human environmental exposure and, thus, have significant implications for human carcinogenesis. PMID- 10676628 TI - High frequency of K-ras mutations in biliary duct carcinomas of cases with a long common channel in the papilla of Vater. AB - The frequency of K-ras mutation in biliary duct carcinomas in different locations and the relationship to the form of the junction of the pancreaticobiliary duct (JPBD) are not understood clearly. These points were investigated in the present study. Thirty-seven biliary duct carcinomas in patients without anomalous JPBD were investigated for K-ras mutations. Regarding location, 12 were hilar, 4 in the upper, 11 in the middle, and 10 in the lower portion of the duct. Furthermore, with 14 cases for which the form of the JPBD could be confirmed by endoscopic retrograde cholangiopancreatography or postoperative cholangiopancreatography, division was made into two types: those with a long common channel (>5 mm) in the papilla of Vater (type 1, n = 4), and the other with a shorter or nonapparent common channel (type 2, n = 10). The overall frequency of K-ras mutation was 30%, the incidence gradually increasing from upper to lower regions. K-ras mutations were significantly more frequent in biliary duct carcinomas associated with long common channels (P < 0.05). These results suggest that a long common channel may bear a relation to K-ras mutations in biliary duct carcinogenesis, presumably through its influence on pancreatic juice regurgitation. PMID- 10676629 TI - Photofrin photodynamic therapy can significantly deplete or preserve oxygenation in human basal cell carcinomas during treatment, depending on fluence rate. AB - At high fluence rates in animal models, photodynamic therapy (PDT) can photochemically deplete ambient tumor oxygen through the generation of singlet oxygen, causing acute hypoxia and limiting treatment effectiveness. We report that standard clinical treatment conditions (1 mg/kg Photofrin, light at 630 nm and 150 mW/cm2), which are highly effective for treating human basal cell carcinomas, significantly diminished tumor oxygen levels during initial light delivery in a majority of carcinomas. Oxygen depletion could be found during at least 40% of the total light dose, but tumors appeared well oxygenated toward the end of treatment. In contrast, initial light delivery at a lower fluence rate of 30 mW/cm2 increased tumor oxygenation in a majority of carcinomas. Laser treatment caused an intensity- and treatment time-dependent increase in tumor temperature. The data suggest that high fluence rate treatment, although effective, may be inefficient. PMID- 10676630 TI - Differences in estrogen receptor alpha variant messenger RNAs between normal human breast tissue and primary breast carcinomas. AB - We evaluated the differences in prevalence and functional activity of human estrogen receptor alpha (hER) variant mRNA between 21 normal breast tissues and 41 primary breast carcinomas using a functional assay in yeast for the hER First, we found that the presence of wild-type hER, relative to the total amount of hER, differs markedly (P < 0.0001) between normal breast tissue (median, 85% wild-type hER) and breast tumors (median, 74% wild-type hER). Second, the hER variants with altered function that are present in normal breast tissue are mainly one-exon deleted splicing variants (median, 100%), whereas in breast tumors only half of all variants lack just one single exon (median, 50%; P < 0.0001). Our results suggest that hER-dependent estrogen responsiveness of breast tissue may change during tumor outgrowth, indicating that specific hER variants may play a role in breast cancer development or progression. PMID- 10676631 TI - High-activity microsomal epoxide hydrolase genotypes and the risk of oral, pharynx, and larynx cancers. AB - Human microsomal epoxide hydrolase (mEH), encoded by the EPHX1 gene, is involved in the metabolism of tobacco carcinogens. We investigated the effect of exon 3 and 4 polymorphisms of the EPHX1 gene in 121 patients with cancers of the oral cavity/pharynx, 129 patients with cancer of the larynx, and 172 non-cancer controls, all Caucasian regular smokers. The potential modifying role of previously analyzed GSTM1, GSTM3, and GSTP1 genotypes was also examined. Compared with the putative low-activity genotypes, odds ratios (ORs) associated with predicted intermediate and high mEH activity genotypes were significantly increased for oropharyngeal cancers [OR = 1.8; 95% confidence interval (CI) = 1.0 3.3; and OR = 2.1; 95% CI = 1.0-4.5, respectively; P(trend) = 0.03] and laryngeal cancers (OR = 1.7; 95% CI = 1.0-3.1; and OR = 2.4; 95% CI = 1.1-5.1, respectively; P(trend) = 0.02). Moreover, a positive interaction was found between mEH activity and GSTM3 genotype for laryngeal cancer. The combined EPHX1 high activity-associated genotype and GSTM3 (AB or BB) genotype conferred a 13.1 fold risk (95% CI = 3.5-48.4) compared with the concurrent presence of the EPHX1 low activity-associated genotype and the GSTM3 AA genotype. Thus, EPHX1 polymorphisms may be one of the factors of importance in susceptibility to smoking-related cancers of the upper aerodigestive tract. PMID- 10676632 TI - Methylation of the human telomerase gene CpG island. AB - The acquisition of expression of hTERT, the catalytic subunit of the telomerase enzyme, seems to be an essential step in the development of a majority of human tumors. However, little is known about the mechanisms preventing telomerase gene expression in normal and transformed cells that do not express hTERT. Using a methylation-specific PCR-based assay, we have found that the CpG island associated with the hTERT gene is unmethylated in telomerase-negative primary tissues and nonimmortalized cultured cells, indicating that mechanisms independent of DNA methylation are sufficient to prevent hTERT expression. The hTERT CpG island is methylated in many telomerase-negative and telomerase positive cultured cells and tumors, but the extent of methylation did not correlate with expression of hTERT. Demethylation of DNA with 5-azacytidine in two cell lines induced expression of hTERT, suggesting that DNA methylation can contribute to hTERT repression in some cells. Together, these data show that the hTERT CpG island can undergo cytosine methylation in cultured cells and tumors and that DNA methylation may contribute to the regulation of the hTERT gene, but that CpG island methylation is not responsible for repressing hTERT expression in most telomerase-negative cells. PMID- 10676633 TI - Constitutive activation of cyclin B1-associated cdc2 kinase overrides p53 mediated G2-M arrest. AB - Recent studies have suggested that p53 regulates the G2 checkpoint in the cell cycle and that this function is required for the maintenance of genomic integrity. In this study, we investigated a regulatory role of p53 specifically in G2-M transition. Human bladder carcinoma cells lacking functional p53 were synchronized at G1-S, which is preceded by p53-mediated G1 arrest. p53 expression in the synchronized cells was induced by infection with a recombinant adenovirus that encodes p53. After release from the G1-S arrest, the cells progressed to S phase and G2 but failed to enter mitosis. Biochemical analysis showed that p53 inhibits cell cycle-dependent expression of cdc2 and cyclin B1 and consequently inhibits cdc2 kinase. The role of cyclin B1-associated cdc2 kinase in p53 mediated G2-M arrest was further investigated by expression of both cyclin B1 and cdc2AF, in which inhibitory phosphorylation sites were substituted. The cells expressing both cdc2AF and cyclin B1 showed a constitutive activation of cdc2 kinase during cell cycle progression and passed through G2-M regardless of p53 expression. Therefore, inactivation of cdc2 kinase through cdc2 and cyclin B1 repression is an essential step in p53-mediated G2-M arrest. PMID- 10676634 TI - No evidence of Peutz-Jeghers syndrome gene LKB1 involvement in left-sided colorectal carcinomas. AB - LKB1 serine/threonine kinase is a gene for Peutz-Jeghers cancer predisposition syndrome. Most studies have detected a low frequency of LKB1 defects in sporadic cancer. A notable exception is a recent report describing frequent, mostly missense type, LKB1 mutations in Korean distal colorectal tumors. To clarify the role of LKB1 in colon cancer, we scrutinized 50 left-sided Korean and Finnish specimens. No somatic mutations were found. The seven Korean somatic missense mutations reported previously were functionally analyzed, and five were found not to alter LKB1 kinase activity. One of these changes was found to be a germ-line polymorphism. LKB1 involvement in distal colorectal cancer is not common. PMID- 10676635 TI - Genomic imbalances including amplification of the tyrosine kinase gene JAK2 in CD30+ Hodgkin cells. AB - Comparative genomic hybridization was applied for a comprehensive screening of frequently occurring net gains and losses of chromosomal subregions in small populations of CD30+ Hodgkin cells and their morphological variants. In 12 Hodgkin's lymphomas, recurrent gains were detected on chromosomal arms 2p, 9p, and 12q (in six, four, and five tumors, respectively) and distinct high-level amplifications were identified on chromosomal bands 4p16, 4q23-q24, and 9p23-p24. In Hodgkin cells with 9p23-p24 amplification, fluorescence in situ hybridization revealed an increased copy number of chromosomal sequences spanning the tyrosine kinase gene JAK2. Several of the imbalances described, in particular a gain in chromosomal arm 9p that includes JAK2 amplification, are similar to the genomic changes detected in primary mediastinal B-cell lymphoma. PMID- 10676636 TI - Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to exert potent cytotoxic activity against many tumor cell lines but not against normal cells. It has been hypothesized that this difference in TRAIL sensitivity between normal and transformed cells might be due to the expression of the non death-inducing TRAIL receptors (TRAIL-R) TRAIL-R3 and TRAIL-R4, presumably by competition for limited amounts of TRAIL. To assess the regulation of resistance versus sensitivity to TRAIL in primary as well as transformed keratinocytes, we examined TRAIL sensitivity, TRAIL receptor expression, and intracellular signaling events induced by TRAIL. Although TRAIL induced apoptosis in primary as well as transformed keratinocytes, a marked difference in sensitivity could be observed with primary keratinocytes (PK) being 5-fold less sensitive to TRAIL than transformed keratinocytes (TK). Yet both cell types exhibited similar TRAIL receptor surface expression, suggesting that expression of TRAIL-R3 and TRAIL-R4 may not be the main regulator of sensitivity to TRAIL. Biochemical analysis of the signaling events induced by TRAIL revealed that PK could be sensitized for TRAIL and, similarly, for TRAIL-R1- and TRAIL-R2-specific apoptosis by pretreatment of the cells with cycloheximide (CHX). This sensitization concomitantly resulted in processing of caspase-8, which did not occur in TRAIL resistant PK. These data indicate that an early block of TRAIL-induced apoptosis was present in PK compared with TK or PK treated with CHX. Interestingly, cellular FLICE inhibitory protein (cFLIP) levels, high in PK and low in TK and several other squamous cell carcinoma cell lines, decreased rapidly after treatment of PK with CHX, correlating with the increase in TRAIL sensitivity and caspase-8 processing. Furthermore, ectopic expression of cFLIP long (cFLIP(L)) in TK by transfection with a cFLIP(L) expression vector resulted in resistance to TRAIL-mediated apoptosis of these cells. Thus, our results demonstrate that TRAIL sensitivity in PK is primarily regulated at the intracellular level rather than at the receptor level. PMID- 10676637 TI - Down-regulation of HER2/neu expression induces apoptosis in human cancer cells that overexpress HER2/neu. AB - The HER2/neu oncogene is overexpressed in a significant fraction of human tumors; such overexpression is thought to play a role in the aberrant proliferation of cancer cells. The effects of HER2/neu-specific phosphorothioate antisense oligodeoxyribonucleotides on HER2/neu expression, tumor cell proliferation, and activation of apoptotic cell death pathways have been examined. Antisense treatment down-regulates HER2/neu expression in a dose-dependent and sequence specific manner. HER2/neu antisense treatment specifically inhibits the growth of tumor lines that overexpress HER2/neu, but it has little effect on the growth of tumor cells that express low levels of HER2/neu. Down-regulation of HER2/neu expression is not only cytostatic, but it also results in the activation of apoptotic cell death pathways in cells that overexpress HER2/neu. These results suggest that, in addition to stimulating tumor cell proliferation, HER2/neu overexpression in cancer cells acts as an antiapoptotic cell survival factor. PMID- 10676638 TI - An indolocarbazole inhibitor of human checkpoint kinase (Chk1) abrogates cell cycle arrest caused by DNA damage. AB - Many cancer therapies cause DNA damage to effectively kill proliferating tumor cells; however, a major limitation of current therapies is the emergence of resistant tumors following initial treatment. Cell cycle checkpoints are involved in the response to DNA damage and specifically prevent cell cycle progression to allow DNA repair. Tumor cells can take advantage of the G2 checkpoint to arrest following DNA damage and avoid immediate cell death. This can contribute to acquisition of drug resistance. By abrogating the G2 checkpoint arrest, it may be possible to synergistically augment tumor cell death induced by DNA damage and circumvent resistance. This requires an understanding of the molecules involved in regulating the checkpoints. Human Chk1 is a recently identified homologue of the Schizosaccharomyces pombe checkpoint kinase gene, which is required for G2 arrest in response to DNA damage. Chk1 phosphorylates the dual specificity phosphatase cdc25C on Ser-216, and this may be involved in preventing cdc25 from activating cdc2/cyclinB and initiating mitosis. To further study the role of Chk1 in G2 checkpoint control, we identified a potent and selective indolocarbazole inhibitor (SB-218078) of Chk1 kinase activity and used this compound to assess cell cycle checkpoint responses. Limited DNA damage induced by gamma-irradiation or the topoisomerase I inhibitor topotecan was used to induce G2 arrest in HeLa cells. In the presence of the Chk1 inhibitor, the cells did not arrest following gamma-irradiation or treatment with topotecan, but continued into mitosis. Abrogation of the damage-arrest checkpoint also enhanced the cytotoxicity of topoisomerase I inhibitors. These studies suggest that Chk1 activity is required for G2 arrest following DNA damage. PMID- 10676639 TI - Quantitative analysis of interindividual variation of glutathione S-transferase expression in human pancreas and the ambiguity of correlating genotype with phenotype. AB - Analysis of glutathione S-transferases (GSTs) of the alpha, mu, and pi classes by reverse-phase high-performance liquid chromatography and electrospray-ionization mass spectrometry in 43 samples of normal human pancreas demonstrated a wide variation in expression of subunits P1, A1, A2, A4, M1, M2, and M3 and the presence of a novel form designated GST "A5." GSTA2 consisted of three forms that were differentially expressed between individuals in a manner consistent with allelic polymorphism at the hGSTA2 locus. Expression, in terms of microg GST subunit/mg cytosolic protein, varied by 6-15-fold for subunits P1, A2, and M3 and 17-30-fold in the case of GSTs A1 and M2. Less consistently expressed were GSTs M1a, M1b, A4, and A5. Among these, GSTM1 expression (excluding M1-null samples) varied 12-fold between samples, whereas GST A4 and A5 expression varied approximately 50-100-fold between samples, well beyond the range of other subunits, suggesting that their expression is highly inducible. Linear correlations (P < 0.001-0.003) existed between levels of the most consistently expressed GST, GSTP1, and total GSTs, GSTA2 and M3, and in GSTM1-positive samples, between GSTM1, M3, and P1. The correlation between GST subunits P1 and M3 was bimodal according to M1 genotype, reflecting the presence of the regulatory element in hGSTM3*B that is linked with the hGSTM1*A genotype. It is concluded that although a degree of regulation of expression of GSTs occurs in human pancreas, the variability of phenotype is high and might obscure the effects of genetic polymorphisms on individual cancer susceptibility. Interindividual variation of GST expression is, therefore, a factor that should be taken account of in epidemiological studies. PMID- 10676640 TI - Differentiation of rat oval cells after activation of peroxisome proliferator activated receptor alpha43. AB - Peroxisome proliferators (PPs) act as nongenotoxic tumor promoters in rodents. Their hepatocarcinogenicity requires the presence of the PP-activated receptor alpha (PPARalpha); however, the exact role played by this transcription factor in the liver, more precisely in liver cell growth and differentiation, is not known. The aim of this study was to investigate the role of PPARalpha in oval cells, which are considered to be closely related to liver stem cells, act as bipotential progenitors for the two main hepatic lineages, and have been implicated as playing a role in several models of liver carcinogenesis. We studied the PPARalpha-mediated response of primary oval cells isolated from rats fed a choline-deficient ethionine-supplemented diet (CDE diet, a regimen commonly used for the induction of oval cell proliferation in rodents) with or without cotreatment with WY14,643, a prototype PPARalpha-activator. PPARalpha was expressed at relatively low levels in primary oval cells from rats fed the CDE diet alone. In vivo treatment with WY14,643 for 2-6 weeks induced, in the oval cells, the expression of PPARalpha as well as that of the PPARalpha-responsive genes encoding fatty acyl-CoA oxidase and cytochrome P450 4A1. Moreover, the oval cell response to WY14,643 was accompanied by an overall phenotypic modulation toward the hepatocyte lineage. In addition, the PPARalpha activator induced, among the oval cells, a subpopulation of transitional cells showing features of maturing hepatocytes expressing the oncofetal marker, alpha-fetoprotein. These results show that oval cells are responsive to PPs and strongly argue for a role of PPARalpha in the differentiation/maturation of rat oval cells. In the absence of the CDE diet regimen, 9-week treatment with WY14,643 lead to the appearance of a population of large-sized cells somewhat similar to the transitional cells. However, these cells showed little expression of markers of mature hepatocytes, consistent with a block during their maturation process, i.e., they are resistant to PPARalpha-mediated differentiation. Interestingly, the phenotype of these cells resembled that of the cells usually found in neoplastic foci induced by PPs. Our results, together with previous reports, suggest the involvement of oval cells in the hepatocarcinogenicity of PPs. PMID- 10676641 TI - Single-site methylation within the p53 promoter region reduces gene expression in a reporter gene construct: possible in vivo relevance during tumorigenesis. AB - It is not known whether transcriptional suppression by de novo methylation occurs within the promoter region of the p53 gene during multistage tumorigenesis. To address this question, in vivo alterations in the CpG methylation within the rat p53 promoter region were evaluated in control, preneoplastic, and tumor tissue during tumor progression using the folate/methyl-deficient model of hepatocarcinogenesis. Alterations in CpG methylation were found to be site specific and to vary depending on the stage of carcinogenesis. To further explore the effect of site-specific methylation on p53 promoter activity, reporter gene constructs were prepared containing specifically methylated sites within the p53 promoter region, and the transcriptional activity in cultured mammalian cells was determined in a transient transfection assay. Relative to the unmethylated construct as a positive control, single-site methylation at nucleotide (nt) -450, which occurs 216 nt upstream from the 85-nt minimal promoter region, suppressed promoter activity by 85%. In contrast, single-site methylation at nt -179, which occurs within the minimal essential promoter region, suppressed activity by only 20%. The p53 promoter constructs containing the singly methylated CpG site at nt 450 were then reevaluated for processive changes in methylation status 48 h after transfection, during maximum suppression of promoter activity. Restriction analysis with methylation-sensitive enzymes revealed that de novo methylation had occurred after transfection at previously unmethylated sites. These findings suggest that nt -450 may constitute a critical site for initiation of de novo methylation and processive spreading of methylation associated with transcriptional inactivation of the p53 gene. Furthermore, the results suggest a possible alternative mechanism for the silencing of the p53 gene in tumors that do not have p53 mutations. PMID- 10676642 TI - Transgenic mice overexpressing protein kinase C epsilon in their epidermis exhibit reduced papilloma burden but enhanced carcinoma formation after tumor promotion. AB - To determine the role that protein kinase C epsilon (PKCepsilon) may play in skin growth, differentiation, and tumor promotion, transgenic mice were generated that overexpressed an epitope-tagged protein kinase C epsilon (T7-PKCepsilon) in their epidermis using the human keratin 14 promoter. Three independent mouse lines that overexpressed the T7-PKCepsilon in their epidermis were produced. The three independent lines 206, 224, and 215 exhibited a 3-, 6-, and 18-fold elevation, respectively, in the level of PKCepsilon immunoreactive protein. Line 215 exhibited a 19-fold greater phosphatidylserine and 12-O-tetradecanoylphorbol-13 acetate (TPA) stimulated kinase activity than line 224. Line 206 exhibited a low basal T7-PKCepsilon activity, which failed to be stimulated by phosphatidylserine and TPA. All of the line 215 transgenic mice (F0 to the F2 generation) displayed phenotypic changes in the skin. The phenotypic changes progressed gradually, starting around 4-5 months of age, with mild dryness of the tail accompanied by hair loss and inflammation at the base of the tail. Hyperproliferation and ulceration of the affected regions were observed around 7-8 months of age. The hyperproliferative epidermis from the affected regions exhibited an expansion of the suprabasal epidermal cells. Inflammation and/or ulceration were also observed in the dorsal skin, the ears, and around the eyes. The line 215 mice, which expressed the highest level of PKCepsilon, were evaluated for sensitivity to mouse skin tumor promotion by TPA. Tumors were elicited by the initiation (7,12 dimethylbenz[a]anthracene, 100 nmol)-promotion (TPA, 5 nmol/twice weekly) protocol. The papilloma burden was reduced by 95-96% for male and female T7 PKCepsilon mice compared to wild-type controls. However, carcinomas developed rapidly in the T7-PKCepsilon mice treated with 7, 12-dimethylbenz[a]anthracene and TPA. These carcinomas appeared to form independently of prior papilloma development. These results demonstrate that PKCepsilon is an important regulator of skin tumor development. PMID- 10676643 TI - Cell proliferation induced by triiodothyronine in rat liver is associated with nodule regression and reduction of hepatocellular carcinomas. AB - Previous studies have demonstrated that short-term treatment with peroxisome proliferators decreased the size and number of gamma-glutamyl transpeptidase or placental glutathione S-transferase (GSTP)-positive hepatic hyperplastic lesions. In this study, we have examined the effect of the hormone triiodothyronine (T3), which, similarly to peroxisome proliferators, is a strong liver mitogen and a ligand of nuclear receptors, on the growth of GSTP-positive nodules generated by the resistant hepatocyte model and on the development of hepatocellular carcinoma. Hepatic hyperplastic nodules were induced in male Fischer rats by a single dose (150 mg/kg) of diethylnitrosamine, followed by a 2-week exposure of the animals to 2-acetylaminofluorene and partial hepatectomy. Nine weeks after diethylnitrosamine administration, rats were switched to a diet containing 4 mg/kg T3 for 1 week (experiment 1) and sacrificed during T3 feeding or were exposed to seven cycles of T3-supplemented diet (1 week/month per 7 months), and sacrificed 6 months after the last cycle (experiment 2). Results showed that T3 treatment for 1 week caused a 70% reduction in the number of GSTP-positive nodules (14/cm2 in T3-fed rats versus 44/cm2 of control animals), as well as GSTP positive area (12% versus 43% of controls). Reduction in the number of GSTP positive nodules observed 1 week after T3 feeding was associated with a strong increase in the labeling index of enzyme-altered nodules compared with that of controls (labeling index was 64 and 31%, respectively). No significant differences in the apoptotic index were observed between the two groups. Results from experiment 2 did reveal that although rats treated with diethylnitrosamine + 2-acetylaminofluorene developed 100% hepatocellular carcinoma and 33% of them showed lung metastasis, only 50% of rats exposed to repeated cycles of triiodothyronine developed hepatocellular carcinoma with no lung metastasis. This study indicates that cell proliferation per se might not necessarily represent a promoting condition for putative preneoplastic lesions and demonstrates an anticarcinogenic effect of T3. PMID- 10676644 TI - Telomerase activity and telomere length in acute and chronic leukemia, pre- and post-ex vivo culture. AB - We studied telomerase regulation and telomere length in hematopoietic progenitor cells from peripheral blood and bone marrow from patients with acute and chronic leukemia and myeloproliferative diseases. CD34+ cells from a total of 93 patients with either acute myeloid leukemia (AML; n = 25), chronic myeloid leukemia (CML; n = 21), chronic lymphocytic leukemia (CLL; n = 18), polycythemia vera (PV; n = 16), or myelodysplastic syndromes (MDS; n = 13) were analyzed before and in 19 patients after ex vivo expansion in the presence of multiple cytokines (kit ligand, interleukin-3, interleukin-6, and granulocyte colony-stimulating factor plus erythropoietin). Compared with hematopoietic progenitor cells from normal donors (n = 108), telomerase activity (TA) was increased 2- to 5-fold in chronic phase (CP)-CML, CLL, PV, and MDS. In AML, accelerated phase (AP) and blastic phase (BP)-CML, basal TA was 10- to 50-fold higher than normal. TA of CP-CML CD34+ cells was up-regulated within 72 h of ex vivo culture, peaked after 1 week, and decreased below detection after 2 weeks. In contrast, TA in AP/BP-CML and AML CD34+ cells was down-regulated after 1 week of culture and decreased further thereafter. The expansion potential of CD34+ cells from patients with leukemia was considerably decreased compared with CD34+ cells from normal donors. The average expansion of cells from leukemic individuals was 6.5-, 2.3-, 0.6-, and 0.2-fold in weeks 1, 2, 3, and 4, respectively, whereas expansion of normal cells was 5- to 15-fold higher. In serial expansion culture, a median telomeric loss of 0.7 kbp was observed during 3-4 weeks of expansion. Our results demonstrate that up-regulation of telomerase is similar in CD34+ cells from CP-CML, CLL, PV, and MDS patients and in normal hematopoietic cells during the first week of culture, whereas in AML and AP/BP-CML, telomerase is high at baseline and down-regulated during expansion culture. High levels of telomerase in leukemic progenitors at baseline may be a feature of both the malignant phenotype and rapid cycling. Telomerase down-regulation during culture of leukemic cells may be due to the decreased expansion potential or repression of normal hematopoiesis, or in AML it may be due to the partial differentiation of AML cells, shown previously to be associated with loss of TA. Telomere shortening during ex vivo expansion correlated with low levels of TA, particularly in chronic leukemic and MDS progenitors where telomerase was insufficient to protect against telomere bp loss during intense proliferation. PMID- 10676645 TI - Altered hepatic gluconeogenesis during L-alanine infusion in weight-losing lung cancer patients as observed by phosphorus magnetic resonance spectroscopy and turnover measurements. AB - Profound alterations in host metabolism in lung cancer patients with weight loss have been reported, including elevated phosphomonoesters (PMEs) as detected by 31P magnetic resonance spectroscopy (MRS). In healthy subjects, infusion of L alanine induced significant increases in hepatic PMEs and phosphodiesters (PDEs) due to rising concentrations of 3-phosphoglycerate and phosphoenolpyruvate, respectively. The aim of the present study was to monitor these changes in the tumor-free liver of lung cancer patients during L-alanine infusion by means of simultaneous 31P MRS and turnover measurements. Twenty-one lung cancer patients without liver metastases with (CaWL) or without weight loss (CaWS), and 12 healthy control subjects were studied during an i.v. L-alanine challenge of 1.4 2.8 mmol/kg followed by 2.8 mmol/kg/h for 90 min. Plasma L-alanine concentrations increased during alanine infusion, from 0.35-0.37 mM at baseline to 5.37 +/- 0.14 mM in the CaWL patients, 6.67 +/- 0.51 mM in the CaWS patients, and 8.47 +/- 0.88 mM in the controls (difference from baseline and between groups during alanine infusion, all P < 0.001). Glucose turnover and liver PME levels at baseline were significantly elevated in the CaWL patients. Alanine infusion increased whole body glucose turnover by 8 +/- 3% in the CaWS patients (P = 0.03), whereas no significant change occurred in the CaWL and controls. PME levels increased by 50 +/- 16% in controls (area under the curve, P < 0.01) and by 87 +/- 31% in the CaWS patients (P < 0.05) after 45-90 min. In contrast, no significant changes in PME levels were observed in the CaWL patients. Plasma insulin concentrations increased during L-alanine infusion in all groups to levels that were lower in the CaWL patients than in the CaWS patients and controls (P < 0.05). In lung cancer patients, but not in controls, changes in PME and PDE levels during alanine infusion were inversely correlated with their respective baseline levels (r = -0.82 and -0.86, respectively; P < 0.001). In addition, changes in PMEs during alanine infusion in lung cancer patients were inversely correlated with the degree of weight loss (r = -0.54; P < 0.05). This study demonstrates the presence of major alterations in the pathway of hepatic gluconeogenesis in weight losing lung cancer patients, as shown by elevated glucose flux before and during L-alanine infusion, and by the increased PME and PDE levels, which reflect accumulation of gluconeogenic intermediates in these patients. Weight-stable lung cancer patients show accelerated increases in PME and PDE levels during L-alanine infusion, suggesting enhanced induction of the gluconeogenic pathway. Our results suggest altered gluconeogenic enzyme activities and elevated alanine uptake within the livers of weight-losing/weight-stable lung cancer patients. PMID- 10676646 TI - Imaging brain tumor proliferative activity with [124I]iododeoxyuridine. AB - Iododeoxyuridine (IUdR) uptake and retention was imaged by positron emission tomography (PET) at 0-48 min and 24 h after administration of 28.0-64.4 MBq (0.76 1.74 mCi) of [124I]IUdR in 20 patients with brain tumors, including meningiomas and gliomas. The PET images were directly compared with gadolinium contrast enhanced or T2-weighted magnetic resonance images. Estimates for IUdR-DNA incorporation in tumor tissue (Ki) required pharmacokinetic modeling and fitting of the 0-48 min dynamically acquired data to correct the 24-h image data for residual, nonincorporated radioactivity that did not clear from the tissue during the 24-h period after IUdR injection. Standard uptake values (SUVs) and tumor:brain activity ratios (Tm:Br) were also calculated from the 24-h image data. The Ki, SUV, and Tm/Br values were related to tumor type and grade, tumor labeling index, and survival after the PET scan. The plasma half-life of [124I]IUdR was short (2-3 min), and the arterial plasma input function was similar between patients (48 +/- 12 SUV*min). Plasma clearance of the major radiolabeled metabolite ([124I]iodide) varied somewhat between patients and was markedly prolonged in one patient with renal insufficiency. It was apparent from our analysis that a sizable fraction (15-93%) of residual nonincorporated radioactivity (largely [124I]iodide) remained in the tumors after the 24-h washout period, and this fraction varied between the different tumor groups. Because the SUV and Tm:Br ratio values reflect both IUdR-DNA incorporated and exchangeable nonincorporated radioactivity, any residual nonincorporated radioactivity will amplify their values and distort their significance and interpretation. This was particularly apparent in the meningioma and glioblastoma multiforme groups of tumors. Mean tumor Ki values ranged between 0.5 +/- 0.9 (meningiomas) and 3.9 +/- 2.3 microl/min/g (peak value for glioblastoma multiforme, GBM). Comparable SUV and Tm:Br values at 24 h ranged from 0.13 +/- 0.03 to 0.29 +/- 0.19 and from 2.0 +/- 0.6 to 6.1 +/- 1.5 for meningiomas and peak GBMs, respectively. Thus, the range of values was much greater for Ki (approximately 8-fold) compared with that for SUV (approximately 2.2-fold) and Tm:Br (approximately 3-fold). The expected relationships between Ki, SUV, and Tm:Br and other measures of tumor proliferation (tumor type and grade, labeling index, and patient survival) were observed. However, greater image specificity and significance of the SUV and Tm:Br values would be obtained by achieving greater washout and clearance of the exchangeable fraction of residual (background) radioactivity in the tumors, i.e., by increased hydration and urinary clearance and possibly by imaging later than 24 h after [124I]IUdR administration. PMID- 10676647 TI - The urokinase system of plasminogen activation and prognosis in 2780 breast cancer patients. AB - The antigen levels of components of the urokinase-type plasminogen activator (uPA) system of plasminogen activation are correlated with prognosis in several types of cancers, including breast cancer. In the present study involving 2780 patients with primary invasive breast cancer, we have evaluated the prognostic importance of the four major components of the uPA system [uPA, the receptor uPAR (CD87), and the inhibitors PAI-1 and PAI-2]. The antigen levels were determined by ELISA in cytosols prepared from primary breast tumors. The levels of the four factors significantly correlated with each other; the Spearman rank correlation coefficients (r(s)) ranged from 0.32 (between PAI-2 and PAI-1 or uPAR) to 0.59 (between uPA and PAI-1). The median duration of follow-up of patients still alive was 88 months. In the multivariate analyses for relapse-free survival (RFS) and overall survival (OS), we defined a basic model including age, menopausal status, tumor size and grade, lymph node status, adjuvant therapy, and steroid hormone receptor status. uPA, uPAR, PAI-1, and PAI-2 were considered as categorical variables, each with two cut points that were established by isotonic regression analysis. Compared with tumors with low levels, those with intermediate and high levels showed a relative hazard rate (RHR) and 95% confidence interval (95% CI) of 1.22 (1.02-1.45) and 1.69 (1.39-2.05) for uPA, and 1.32 (1.14-1.54) and 2.17 (1.74-2.70) for PAI-1, respectively, in multivariate analysis for RFS in all patients. Compared with tumors with high PAI-2 levels, those with intermediate and low levels showed a poor RFS with a RHR (95% CI) of 1.30 (1.14-1.48) and 1.76 (1.38-2.24), respectively. Similar results were obtained in the multivariate analysis for OS in all patients. Furthermore, uPA and PAI-1 were independent predictive factors of a poor RFS and OS in node-negative and node-positive patients. PAI-2 also added to the multivariate models for RFS in node-negative and node-positive patients, and in the analysis for OS in node-negative patients. uPAR did not further contribute to any of the multivariate models. A prognostic score was calculated based on the estimates from the final multivariate model for RFS. Using this score, the difference between the highest and lowest 10% risk groups was 66% in the analysis for RFS at 10 years and 61% in the analysis for OS. Moreover, separate prognostic scores were calculated for node-negative and node-positive patients. In the 10% highest risk groups, the proportion of disease free patients was only 27 +/- 6% and 9 +/- 3% at 10 years for node-negative and node-positive patients, respectively. These proportions were 86 +/- 4% and 61 +/- 6% for the corresponding 10% lowest risk groups of relapse. We conclude that several components of the uPA system are potential predictors of RFS and OS in patients with primary invasive breast cancer. Knowledge of these factors could be helpful to assess the individual risk of patients, to select various types of adjuvant treatment and to identify patients who may benefit from targeted therapies that are currently being developed. PMID- 10676648 TI - Substantially reduced risk of cancer of the aerodigestive tract in subjects with variant--463A of the myeloperoxidase gene. AB - Myeloperoxidase (MPO), an enzyme that is highly expressed in neutrophil leukocytes, transforms precarcinogens such as benzo(a)pyrene and aromatic amines to highly reactive intermediates. A G/A polymorphism located 463 bp upstream of exon 1 in the promoter region strongly reduces MPO mRNA expression. In a matched case-control study, 196 lung cancer, 245 laryngeal cancer, and 255 pharyngeal cancer patients from the Berlin area were investigated for frequency of the G 463A polymorphism by PCR/RFLP, using AciI. They were matched by age and gender to hospital patients without known malignancies. Moreover, 270 healthy volunteers were genotyped, obtaining 61.1% of individuals with MPO genotype -463G/G, 34.8% of individuals with genotype G/A, and 4.1% of individuals with genotype A/A. In lung and laryngeal cancer patients, but not in pharyngeal cancer patients, mutant genotypes were significantly less frequent. Crude odds ratios for carriers of one or two A alleles, compared to wild-type G/G, were 0.58 [95% confidence interval (CI), 0.38-0.88; P = 0.011] for lung cancer patients, 0.63 (95% CI, 0.43-0.92; P = 0.017) for laryngeal cancer patients, and 0.82 (95% CI, 0.57-1.17; P = 0.27) for pharyngeal cancer patients. The relative risks, adjusted for age, gender, and extent of cigarette smoking were 0.47 (95% CI, 0.28-0.79; P = 0.004), 0.66 (95% CI, 0.44-1.01; P = 0.054), and 0.75 (95% CI, 0.51-1.12; P = 0.16) for lung, larynx, and pharyngeal cancer, respectively. Strikingly, relative risk for carriers of -463A among adenocarcinoma of the lung was 0.24 (95% CI, 0.10-0.58; P = 0.002). Two cases with larynx cancer, one case with lung cancer, and one reference subject displayed novel G/A mutations at -297 nucleotide and -296 nucleotide, destroying a constitutive AciI cleavage site. Our data finally suggest that the MPO -463A variant is a protective factor in the etiology of lung and larynx cancer, but possibly not of pharyngeal cancer. PMID- 10676649 TI - Manumycin enhances the cytotoxic effect of paclitaxel on anaplastic thyroid carcinoma cells. AB - Despite the current multimodal approach to treatment of anaplastic thyroid cancer (ATC), the prognosis for patients with the disease is poor. New effective therapy for ATC is desperately needed. Thus, we investigated the effects of manumycin (a farnesyl:protein transferase inhibitor), alone and in combination with other drugs frequently used to treat ATC, in six human ATC cell lines: ARO, C643, DRO, Hth-74, KAT-4, and KAT-18. By means of a formazan dye-based spectrophotometric assay of cell viability and light microscopy, manumycin was shown to decrease the number of viable cells in all six of the cell lines though to a lesser degree in DRO and C643 cells than in ARO, Hth-74, KAT-4, and KAT-18 cells. In combination, manumycin enhanced the effect of paclitaxel in all six of the cell lines. The mechanism of cell death was investigated by measuring caspase-3 activity, immunoblotting with anti-poly-(ADP-ribose)polymerase (PARP) antibody and electrophoresis of DNA. After an 18-h incubation, manumycin plus paclitaxel caused enhanced activation of caspase-3 activity, cleavage of PARP into Mr 89,000 and 28,000 fragments, and internucleosomal fragmentation of DNA (all of which are characteristic of apoptotic cell death). In contrast, neither manumycin alone, paclitaxel alone, doxorubicin alone, nor doxorubicin plus manumycin produced significant specific cleavage of PARP and internucleosomal DNA fragmentation after 18 h of incubation. The in vivo effect and toxicity of combined manumycin and paclitaxel treatments were evaluated in a nude mouse xenograft model using ARO and KAT-4 cells. Drugs were injected i.p. on days 1 and 3 of a 7-day cycle for three cycles. Both manumycin (7.5 mg/kg/dose) and paclitaxel (20 mg/kg/dose) had significant inhibitory effects on tumor growth. Combined manumycin and paclitaxel treatments seemed as effective as manumycin against ARO cells and more effective than either manumycin or paclitaxel alone against KAT-4 cells. No significant morbidity or mortality was caused by the treatments. In conclusion, manumycin can inhibit the growth of ATC both in vitro and in vivo. Manumycin plus paclitaxel has enhanced cytotoxic effects and increased apoptotic cell death in ATC cells in vitro compared with either drug by itself. The combination of manumycin and paclitaxel is also effective in vivo with no significant toxicity observed. The lack of synergy observed in this in vivo experiment may be due to a ceiling effect, and further experimentation is warranted to ascertain the optimal way to combine these two agents for maximal therapeutic effects. PMID- 10676650 TI - Expression of endogenously activated secreted or cell surface carboxypeptidase A sensitizes tumor cells to methotrexate-alpha-peptide prodrugs. AB - Methotrexate (MTX) is one of the most commonly used agents in the treatment of solid malignancies; however, the toxicities of MTX to bone marrow and gastrointestinal tract complicate this therapy. We, therefore, propose a gene dependent enzyme prodrug therapy to limit these toxicities by localizing the production of MTX to the site of the tumor. The combination of MTX-alpha-peptide prodrugs, which cannot be internalized by the cellular reduced folate carrier, with carboxypeptidase A (CPA), which can remove the blocking peptide, has been demonstrated previously in vitro using antibody-dependent enzyme prodrug therapy. CPA is normally synthesized as a zymogen that is inactive without proteolytic removal of its propeptide by trypsin. Therefore, to adapt this system to gene dependent enzyme prodrug therapy, a mutant form of CPA was engineered, CPA(ST3), that does not require trypsin-dependent zymogen cleavage but is instead activated by ubiquitously expressed intracellular propeptidases. Purification, peptide sequencing, and kinetic analysis indicated that mature CPA(ST3) is structurally and functionally similar to the trypsin-activated, wild-type enzyme. In addition, CPA(ST3)-expressing tumors cells were sensitized to MTX prodrugs in a dose- and time-dependent manner. To limit diffusion of CPA, a cell surface localized form was generated by constructing a fusion protein between CPA(ST3) and the phosphatidylinositol linkage domain from decay accelerating factor. SDS-PAGE and flow cytometric analysis of infected tumor cells indicated that CPA(DAF) was cell surface localized. Finally, after retroviral transduction, this enzyme/prodrug strategy exhibited a potent bystander effect, even when <10% of the cells were transduced, because extracellular production of MTX sensitized both transduced and nontransduced cells. PMID- 10676651 TI - Tumor-targeting chemotherapy by a xanthine oxidase-polymer conjugate that generates oxygen-free radicals in tumor tissue. AB - Xanthine oxidase (XO) mediates anticancer activity because of its ability to generate cytotoxic reactive oxygen species (ROS), including superoxide anion radical and hydrogen peroxide. However, the high binding affinity of XO to blood vessels would cause systemic vascular damage and hence limits the use of native XO in clinical settings. We demonstrate here that chemical conjugation of XO with poly(ethylene glycol) (PEG; the conjugates hereafter referred to as PEG-XO) significantly enhanced the tumor-targeting efficacy and the antitumor activity of XO. By using a succinimide-activated PEG derivative, PEG was conjugated to epsilon-amino groups of lysine residues of XO, which play a crucial role in binding of XO to blood vessels. PEG-XO administered i.v. showed a 2.8-fold higher accumulation in solid tumor compared with that of native XO 24 h after injection, whereas a slight or negligible increase in accumulation of PEG-XO was observed in normal organs. The highest PEG-XO enzyme activity was detected in tumor compared with normal organs or tissues except blood; enzyme activity in tumor was 5.0, 3.9, and 9.4 times higher than that in liver, kidney, and spleen, respectively. Intratumor activity remained high for >48 h. Administration of hypoxanthine, a substrate of XO, at 33 mg/kg body weight i.p. 12 h after the administration of PEG-XO (0.6 unit/mouse, i.v.) resulted in significant suppression of tumor growth (P < 0.001), with no tumor growth even after 52 days. However, either PEG-XO or hypoxanthine alone, or native XO with hypoxanthine, showed no effect on the inhibition of tumor growth under present experimental conditions. These findings suggest that PEG-XO, which accumulates preferentially in tumor tissue, warrants further investigation as a novel anticancer agent. PMID- 10676652 TI - Functional interactions between bile acids, all-trans retinoic acid, and 1,25 dihydroxy-vitamin D3 on monocytic differentiation and myeloblastin gene down regulation in HL60 and THP-1 human leukemia cells. AB - Bile acids were shown previously to inhibit proliferation and to induce monocytic differentiation in HL60 human acute promyelocytic leukemia cells (A. Zimber et al., Int. J. Cancer, 59: 71-77, 1994). In this report, we hypothesized that bile acids may exert a positive cooperativity with two known inducers of leukemic cell differentiation, all-trans retinoic acid and 1,25(OH)2-vitamin D3. Our results provide evidence that bile acids induced the monocytic differentiation of HL60 and THP-1 human leukemia cells exposed to ineffective concentrations of these inducers. The protein kinase C (PKC) inhibitors H-7 (10 and 20 microM) and staurosporine (5 and 20 nM) modulated the effects of bile acids on HL60 cell differentiation. Most interestingly, bile acids are shown herein to down-regulate the expression of the serine protease myeloblastin gene involved in the differentiation of myeloid hematopoietic cells. In agreement with the recent identification of nuclear receptors for bile acids, our data suggest that functional interactions between nuclear bile acid signaling pathways, PKC, and nuclear receptors for retinoic acid and vitamin D3 are involved in the down regulation of the myeloblastin gene and the induction of cell differentiation in human leukemic cells. PMID- 10676653 TI - p21WAF1/CIP1 antisense therapy radiosensitizes human colon cancer by converting growth arrest to apoptosis. AB - Substantial evidence suggests that loss of cellular p21WAF1/CIP1 results in increased apoptotic killing by ionizing radiation. We hypothesized that a p21 antisense (AS) oligodeoxynucleotide (ODN) could be used to sensitize cancer cells to radiotherapy. In vitro treatment of colon cancer cells (HCT116/p21+/+) with p21 AS ODN (200 nM) led to inhibition of radiation-induced p21 expression (>95% inhibition, 0-30 Gy), resulting in a loss of G1 arrest and an enhancement of apoptosis to comparable levels and with similar kinetics to HCT116/p21-/- cells (approximately 60% apoptotic cells at 96 h after 10 Gy). In vivo, p21 AS ODN in combination with radiation (i.p. ODN for 6 days at 20 mg/kg/day and 15 Gy) increased apoptosis in s.c. p21+/+ tumors in nude mice to levels similar to those of p21-/- tumors (2-fold at 24 h postirradiation) and improved radiocurability of p21+/+ tumors to levels comparable to those of p21-/- tumors (p21+/+, two of eight cures versus p21-/-, two of nine cures). Our findings suggest that p21 AS treatment may be a rational approach to improve conventional radiotherapy outcomes. PMID- 10676654 TI - Interleukin-7/B7.1-encoding adenoviruses induce rejection of transplanted but not nontransplanted tumors. AB - Most cancer vaccine trials are based on efficacy studies against transplanted mouse tumors that poorly reflect the clinical situation. We constructed adenoviruses expressing interleukin-7 and B7.1 and tested their therapeutic efficacy after transfer into established transplanted and nontransplanted 3 methylcholanthrene-induced tumors. The adenoviruses efficiently induced rejection of transplanted tumors, leaving behind systemic immunity. Against nontransplanted tumors of similar size, there were almost no therapeutic effects. This result was not due to the site of tumor development, tumor type, general immune suppression, or differences in transduction efficacy. Adenoviral expression of beta galactosidase as a surrogate antigen in nontransplanted tumors induced cytotoxic T cells that were unable to quantitatively reach the tumor site. Based on rigorous mouse models and an effective in situ immunization procedure, it is suggested that cancer vaccines can be effective, if at all, against "minimal residual disease"; additional experimental procedures must be found against established nontransplanted tumors. PMID- 10676655 TI - Targeting of human p53-overexpressing tumor cells by an HLA A*0201-restricted murine T-cell receptor expressed in Jurkat T cells. AB - A potent anti-human (hu) p53 CD8+ CTL response develops in HLA A*0201 transgenic (Tg) mice after immunization with peptides corresponding to HLA A*0201 motifs from hu p53. Mice immunized with the hu P53(149-157) peptide develop a CTL response that is of moderately high affinity and is capable of recognizing hu tumor cells expressing mutated p53. In this report, the mRNAs encoding the predominantly expressed T-cell receptor (TCR) sequences were molecularly cloned from a murine (mu) CTL clone derived from immunized Tg mice, which recognized endogenously processed hu p53 restricted by HLA A*0201. The separate A and B chain TCR cDNAs were transfected in the corresponding TCR A- and B- Jurkat-CD3- mutant T-cell lines, and each rescued CD3 surface expression. Both TCR chains were simultaneously introduced into Jurkat-CD3+ cells, and the transfected Jurkat cells recognized hu T2 cells sensitized with the p53(149-157) CTL epitope but not T2 cells sensitized with a nonspecific CTL epitope. Breast, pancreatic, and sarcoma tumor cell lines, which overexpress endogenous mutated p53, were recognized in the presence of anti-CD28 costimulation, only if they also expressed HLA A*0201. Normal hu fibroblasts established from skin cultures were not recognized. These results represent the first time that a p53-specific TCR capable of recognizing hu cancer cells was heterologously expressed in a naive recipient cell, converting that cell to one recognizing hu tumor cells with mutated p53. This TCR represents a candidate molecule for a genetic strategy in combating hu cancer by an adoptive immunotherapy approach, which uses the strong xenorecognition of hu p53 in mice. PMID- 10676656 TI - Frequent methylation of estrogen receptor in prostate cancer: correlation with tumor progression. AB - Prior studies have shown that the estrogen receptor (ER) gene is down-regulated in prostate cancer, but the mechanism of its inactivation is not known. We hypothesize that inactivation of the ER gene in prostate cancer is through promoter methylation. To test this hypothesis, we investigated the methylation status of the ER gene in prostate cancer cell lines, prostate cancer, and benign prostatic hyperplasia (BPH) tissues samples using the bisulfite genomic sequencing method. Our results show that the ER gene promoter was methylated in 100% (six of six) of the prostate cancer cell lines tested and all were accompanied by loss of ER mRNA expression. Treatment of these cell lines with demethylating agent 5-aza-2'-deoxycytidine restored ER mRNA expression in all of the ER-negative cell lines. In addition, elevated expression of DNA methyltransferase mRNA was found in all of the prostate cancer cell lines. Of the prostate tissue samples analyzed, 60% (6 of 10) in the BPH samples, 80% (8 of 10) in the low-grade cancer samples (grades I and II), and 95% (20 of 21) in the high grade cancer samples (grades III-V) exhibited promoter methylation of the ER gene. The overall methylation levels in the cancer samples were higher than that in the BPH samples. The differences between the high-grade cancer samples and BPH samples were significant at all CpG sites. Only at three CpG sites were the differences significant between the low-grade cancer samples and BPH samples. This study presents the first evidence that ER gene is transcriptionally inactivated by DNA methylation in prostate cancer. Our data suggest that ER may be involved in the pathogenesis of prostate cancer, as well as BPH. PMID- 10676657 TI - Detection of plasma tumor DNA in head and neck squamous cell carcinoma by microsatellite typing and p53 mutation analysis. AB - Recent arguments have suggested that tumor DNA in cancer patients could be found in plasma, but different points remain unclear. Using a series of 117 head and neck squamous cell carcinoma tumors, our goals for this study were: (a) to quantify the amount of plasma DNA; (b) to evaluate the presence of plasma tumor DNA; and (c) to analyze the clinical relevance of tests based on plasma DNA analyses. Low levels of plasma DNA were found in most samples, but all were successfully amplified. Two different methods were used to detect tumor-specific genetic alterations: (a) microsatellite instability at UT5085 with an established sensitivity of 1:500; and (b) p53 mutation screening. Of the 117 tumors typed at UT5085, 65 demonstrated bandshifts (55%). Plasma and tumor DNA a showed similar alteration in only one case among these samples, and the prevalence of tumor DNA in plasma was estimated to be <2% using microsatellite analysis. Tumor DNA was detected in plasma at a higher prevalence (2 of 11 cases) when using p53 mutant allele-specific amplification. These results showed that in plasma, tumor DNA is largely diluted by normal DNA. By comparison with previously published studies, the prevalence of microsatellite alterations in plasma in this series of head and neck squamous cell carcinomas is very low, despite the fact that a large series of tumors was analyzed. To explain this discrepancy, we analyzed the possibility of PCR artifacts as suspected by the presence of loss of heterozygosity in two plasma DNA samples without a similar tumor DNA alteration. When DNA concentrations were under the threshold of detection (<100 ng/ml), we demonstrated that PCR artifacts could occur at random, and, if misinterpreted, these false genetic alterations could artificially enhance the frequency of plasma DNA alterations. This may have been suspected in previously published series, but it has never been discussed before. Microsatellite analysis on plasma DNA is difficult to interpret and can frequently be misleading. Plasma DNA should be analyzed with very sensitive and specific methods such as mutant allele specific amplification, which excludes artifacts but requires specific optimization that is probably not compatible with routine and clinical use. PMID- 10676658 TI - Integrins alpha(v)beta3 and alpha(v)beta5 are expressed by endothelium of high risk neuroblastoma and their inhibition is associated with increased endogenous ceramide. AB - Inhibition of the RGD-binding integrins, alpha(v)beta3 and alpha(v)beta5, prevents endothelial cell anchorage and induces endothelial apoptosis, which results in disruption of tumor angiogenesis and inhibition of tumor growth in animal models. In this study, we demonstrate by immunohistochemical analysis that integrin alpha(v)beta3 was expressed by 61% (mean) of microvessels in high-risk neuroblastomas (stage IV and MYCN-amplified stage III; n = 28) but only by 18% (mean) of microvessels in low-risk tumors (stages I and II and non-MYCN-amplified stage III; n = 12). Integrin alpha(v)beta5 was found on 60% (mean) of microvessels in 21 Stage IV tumors. These data suggest that neuroblastomas may be targeted for antiangiogenic treatment directed against endothelial integrins alpha(v)beta3 and alpha(v)beta5. In cell culture, inhibition of integrin dependent endothelial cell anchorage to vitronectin by RGDfV, an RGD function blocking cyclic peptide, induced apoptosis in bovine brain endothelial cells compared with the control peptide, RADfV (37.5% versus 8.7%, respectively), as detected by chromatin condensation and nuclear fragmentation. Treatment with RGDfV but not with RADfV, which prevented attachment of endothelial cells to vitronectin or fibronectin, was associated with up to a 50% increase in endogenous ceramide, a lipid second messenger that can mediate cell death. Furthermore, exogenous C2-ceramide was cytotoxic to bovine brain endothelial cells and induced activation of C-jun N-terminal kinase (JNK), a MAP kinase that can be activated in stress-induced apoptosis pathways. This suggests that ceramide may function in detachment-induced endothelial cell apoptosis, originating from inhibition of vitronectin binding to integrins such as alpha(v)beta3 and alpha(v)beta5. This is the first report to demonstrate expression of integrins alpha(v)beta3 and alpha(v)beta5 by microvascular endothelium of a childhood tumor and association of their expression with neuroblastoma aggressiveness. Furthermore, our data provide the first suggestion that inhibition of endothelial cell anchorage, resulting from specific blockade of RGD-binding integrins, increases endogenous ceramide, which may contribute to endothelial cell death. PMID- 10676659 TI - Aminopeptidase N is a receptor for tumor-homing peptides and a target for inhibiting angiogenesis. AB - Phage that display a surface peptide with the NGR sequence motif home selectively to tumor vasculature in vivo. A drug coupled to an NGR peptide has more potent antitumor effects than the free drug [W. Arap et al., Science (Washington DC), 279: 377-380, 1998]. We show here that the receptor for the NGR peptides in tumor vasculature is aminopeptidase N (APN; also called CD13). NGR phage specifically bound to immunocaptured APN and to cells engineered to express APN on their surface. Antibodies against APN inhibited in vivo tumor homing by the NGR phage. Immunohistochemical staining showed that APN expression is up-regulated in endothelial cells within mouse and human tumors. In another tissue that undergoes angiogenesis, corpus luteum, blood vessels also expressed APN, but APN was not detected in blood vessels of various other normal tissues stained under the same conditions. APN antagonists specifically inhibited angiogenesis in chorioallantoic membranes and in the retina and suppressed tumor growth. Thus, APN is involved in angiogenesis and can serve as a target for delivering drugs into tumors and for inhibiting angiogenesis. PMID- 10676660 TI - Selective expression and constitutive phosphorylation of SHC proteins [corrected] in the CD34+ fraction of chronic myelogenous leukemias. AB - The BCR/ABL fusion protein is a constitutively active tyrosine kinase that is responsible for the pathogenesis of chronic myelogenous leukemia (CML). Clinically, CML is characterized by a chronic phase (CP) that eventually terminates into a blast crisis (BC). BC transformation is associated with accumulation of CD34+ blasts. We investigated the expression and phosphorylation of Src-homology-2 and collagen-homology domains (SHC) [corrected] proteins in subpopulations of CML primary cells. Shc polypeptides are tyrosine kinase substrates that are constitutively tyrosine-phosphorylated in continuous cell lines of CML origin. High levels of Shc expression were found in the CD34+ cells from CML-BC, CML-CP and normal bone marrow. In contrast, CD34- fractions from CML CP and normal bone marrow expressed low levels of p46Shc. Shc proteins were constitutively phosphorylated in the CD34+ fractions from CML cells (both CP and BC), but not in normal CD34+ cells. These data bear implications for the role of Shc in normal hemopoiesis and CML leukemogenesis: (a) dramatic changes of Shc expression during terminal differentiation of hemopoietic cells adds a further level of regulation to the signal transduction function of Shc; and (b) constitutive Shc tyrosine-phosphorylation in the rare CD34+ cells of CML-CP might contribute to the selection of this subpopulation during the blast crisis transformation of CMLs. PMID- 10676661 TI - Basic fibroblast growth factor confers a less malignant phenotype in MDA-MB-231 human breast cancer cells. AB - Basic fibroblast growth factor (FGF-2) expression is associated with a more differentiated phenotype, earlier stage of disease, and a better prognosis in breast cancer patients. To determine whether expression of FGF-2 can cause a less malignant phenotype, we engineered MDA-MB-231 cells, a highly dedifferentiated, invasive breast cancer cell line, to express different isoforms of FGF-2. Cells expressed either cytoplasmic, nuclear, or a combination of both FGF-2 isoforms. Western blots of 2 M NaCl washes and of conditioned medium demonstrated that these cells did not export FGF-2. Cells expressing FGF-2 had levels of fibroblast growth factor receptors equivalent with those of control cells. Transformation was assayed by anchorage-independent colony formation and tumor formation in athymic mice. All of the constructs expressing various FGF-2 isoforms had a 60 70% reduction in colony formation in soft agar, but only cells expressing the Mr 18,000 FGF-2 isoform formed fewer and smaller tumors in mice. To determine potential mechanisms responsible for a less malignant phenotype, experiments measuring invasion in Matrigel, the secretion of matrix metalloprotease activity and migration in a modified Boyden chamber and in a patch wound motility assay were carried out. Cells expressing the Mr 18,000 cytoplasmic FGF-2 moiety had a 45% decrease in invasion in Matrigel compared to vector-transfected controls. Cells expressing Mr 18,000 FGF-2 had an increase in Mr 97,000 and Mr 48,000 collagenase, demonstrating that the decreased invasive potential was not due to a down-regulation of gelatinolytic or caseinolytic matrix metalloproteinases. However, motility was decreased in both assays, primarily in cells expressing Mr 18,000 FGF-2, whereas exogenous recombinant human FGF-2 had no effect. These studies demonstrate for the first time that FGF-2 expression can cause a less malignant phenotype in breast cancer cells, possibly as a result of decreased motility and invasion. PMID- 10676662 TI - Androgen deprivation of the PC-310 [correction of prohormone convertase-310] human prostate cancer model system induces neuroendocrine differentiation. AB - Neuroendocrine (NE) cells are androgen-independent cells and secrete growth modulating neuropeptides via a regulated secretory pathway (RSP). We studied NE differentiation after androgen withdrawal in the androgen-dependent prostate cancer xenograft PC-310. Expression patterns of chromogranin A, secretogranin III, and prohormone convertase-1 were analyzed at both protein and mRNA level to mark the kinetics of NE differentiation both in vivo and in vitro. PC-310 tumor bearing nude mice were killed at 0, 2, 5, 7, 14, and 21 days postcastration. PC 310C cultures initiated from collagenase-treated tumor tissue could be maintained up to four passages, and androgen-deprivation experiments were performed similarly. PC-310 tumor volumes decreased by 50% in 10 days postcastration. Proliferative activity and prostate-specific antigen (PSA) serum levels decreased to zero postcastration, whereas PSA levels in PC-310C culture media first decreased and subsequently increased after 5 days. In vivo, androgen receptor (AR) expression decreased initially but returned to control level from 5 days postcastration on. CgA, secretogranin III, and secretogranin V expression increased in vivo from 5 days postcastration on. Subsequently, prohormone convertase-1 and peptidyl alpha-amidating monooxygenase as well as the vascular endothelial growth factor were expressed from 7 days postcastration on, and, finally, growth factors such as gastrin-releasing peptide and serotonin were expressed in a small part of the NE cells 21 days postcastration. The PC-310 tumors did not show colocalization of the AR on the NE cells in the tumor residues after 21 days. As in the PC-310 xenograft, NE differentiation was induced and AR expression relapsed after prolonged androgen suppression in PC 310C. For PC-310C cells, this relapse was associated with the secretion of PSA. PC-310C is the first culture of human prostatic cancer cells having the NE phenotype. The PC-310 model system is a potential androgen-dependent model for studying the role of NE cells in the progression of clinical prostate cancer. Androgen deprivation of NE-differentiated prostate cancer may induce the formation of both NE- and AR-positive dormant tumor residues, capable of actively producing NE growth factors via a RSP, possibly leading to hormone refractory disease. PMID- 10676663 TI - Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. AB - A gene related to cell differentiation was identified by differential display as a candidate suppressor of metastases in colon cancer. This gene, with a full length cDNA of 3 kb, is expressed in normal colon and primary colon cancer tissues and cell lines but not in their metastatic counterparts. A GenBank search found that it is identical to a recently cloned gene, differentiation-related gene-1 (Drg-1), isolated from differentiated HT-29 colon cancer cells. Stable transfection of the SW620 metastatic colon cancer cell line with Drg-1 cDNA induced morphological changes consistent with differentiation and up-regulated the expression of several colonic epithelial cell differentiation markers (alkaline phosphatase, carcinoembryonic antigen, and E-cadherin). Moreover, the expression of Drg-1 is controlled by several known cell differentiation reagents, such as ligands of peroxisome proliferator-activated receptor gamma (troglitazone and BRL46593) and of retinoid X receptor (LG268), and histone deacetylase inhibitors (trichostatin A, suberoylanilide hydroxamic acid, and tributyrin). A synergistic induction of Drg-1 expression was seen with the combination of tributyrin and a low dose of 5'-aza-2'-dexoycytidine (100 nM), an inhibitor of DNA methylation. Functional studies revealed that overexpression of Drg-1 in metastatic colon cancer cells reduced in vitro invasion through Matrigel and suppressed in vivo liver metastases in nude mice. We propose that Drg-1 suppresses colon cancer metastasis by inducing colon cancer cell differentiation and partially reversing the metastatic phenotype. PMID- 10676664 TI - A precursor form of prostate-specific antigen is more highly elevated in prostate cancer compared with benign transition zone prostate tissue. AB - Prostate-specific antigen (PSA) is a widely used serum marker for prostate cancer (PCa), but in the critical diagnostic range of 4-10 ng/ml it has limited specificity for distinguishing early PCa from benign prostatic hyperplasia (BPH). PSA in serum is comprised of a variety of both "free" and "complexed" forms that have been used to improve the specificity of PSA for prostate cancer detection. We previously reported that pro PSA (pPSA), the zymogen or precursor form of PSA, is a component of free PSA in the serum of PCa patients. In the current study, we examined prostate tissues to understand the origin and specificity of pPSA. PSA was immuno-affinity purified from matched sets of prostate tissues including peripheral zone cancer (PZ-C); peripheral zone noncancer; and benign tissue from the transition zone (TZ), the primary site of BPH within the prostate. We found that pPSA is differentially elevated in PZ-C, but is largely undetectable in TZ. N-terminal sequencing revealed that the pPSA was comprised primarily of [-2]pPSA and minor levels of [-4]pPSA, containing pro leader peptides of 2 and 4 amino acids, respectively. The median value of pPSA was 3% in PZ-C and 0% (undetectable) in TZ (P < 0.0026). No pPSA was detected in 13 of 18 transition zone specimens (72%), but only 2 of the 18 matched cancer specimens (11%) contained no measurable pPSA. These results demonstrate that pPSA is more highly correlated with prostate cancer than with BPH. The pPSA in serum may represent a more cancer-specific form of PSA that could help distinguish prostate cancer from BPH. PMID- 10676665 TI - Down-regulation of the insulin-like growth factor I receptor by antisense RNA can reverse the transformed phenotype of human cervical cancer cell lines. AB - The insulin-like growth factor I receptor (IGF-IR) plays an essential role in the establishment and maintenance of transformed phenotype, and interference with the IGF-IR pathway by antisense or dominant-negative mutants causes reversal of the transformed phenotype in many rodent and human tumor cell lines. We stably transfected an IGF-IR antisense mRNA expression plasmid into human papillomavirus (HPV)-negative C33a cell line, HPV-16-positive SiHa cell line, and HPV-18 positive HeLa S3 cell line to determine whether the IGF-IR could be a target for cervical cancer cells, especially in the presence of HPV. Approximately 30-80% down-regulation of IGF-IR expression was observed by Western blot in antisense transfected clones. There was a little inhibition in monolayer growth in all cell lines. In C33a cells, wild-type and sense clones formed 92-146 colonies in soft agar after 3 weeks; antisense clones formed <12 colonies. In SiHa cells, wild type and sense clones formed approximately 60 colonies after 5 weeks; antisense clones formed 0-3 colonies. In HeLa S3 cells, wild-type and sense clones formed 218-291 colonies in soft agar after 2 weeks; antisense clones formed 14-160 colonies. There was a good correlation between IGF-IR down-regulation level and inhibition of transformation in soft agar. Tumorigenesis in nude mice was strongly inhibited in HeLa S3 and SiHa clones transfected with the antisense. These results indicate that down-regulation of IGF-IR by antisense RNA can reverse the transformed phenotype of human cervical cancer cells, even when harboring malignant type HPVs. PMID- 10676666 TI - Correspondence re: S. Shintani et al., Intragenic mutation analysis of the human epidermal growth factor receptor (EGFR) gene in malignant human oral keratinocytes. Cancer Res., 59: 4142-4147, 1999. PMID- 10676667 TI - Eleventh annual Pezcoller Symposium: molecular horizons in cancer therapeutics. PMID- 10676668 TI - Dietary supplementation with marine omega-3 fatty acids improve systemic large artery endothelial function in subjects with hypercholesterolemia. AB - OBJECTIVE: This work was undertaken to determine whether dietary supplementation with marine omega-3 fatty acids improve systemic large artery endothelial function in subjects with hypercholesterolemia. BACKGROUND: Marine omega-3 fatty acids improve vascular function, but the underlying mechanism(s) are unclear. We studied the effects of marine omega-3 fatty acids on large artery endothelial function in subjects with hypercholesterolemia. METHODS: Hypercholesterolemic subjects with no other known cause for endothelial dysfunction were recruited to a prospective, placebo-controlled, randomized, double-blind, parallel-group study. Treatment with omega-3 fatty acids at a dose of 4 g/day (n = 15/group) was compared with placebo, at the beginning (day 0) and end (day 120) of a four-month treatment period. Endothelial function was assessed pre- and posttreatment by noninvasive ultrasonic vessel wall tracking of brachial artery flow-mediated dilation (FMD). RESULTS: Treatment with marine omega-3 fatty acids resulted in a significant improvement in FMD (0.05 +/- 0.12 to 0.12 +/- 0.07 mm, p < 0.05) and a significant reduction in triglycerides (2.07 +/- 1.13 to 1.73 +/- 0.95 mmol/liter, p < 0.05), whereas treatment with placebo resulted in no change in FMD (0.03 +/- 0.10 to 0.04 +/- 0.10 mm) or triglycerides (2.29 +/- 2.09 to 2.05 +/- 1.36 mmol/liter) (both p < 0.05 treated compared with control). Responses to sublingual glyceryl trinitrate were unchanged. CONCLUSIONS: Marine omega-3 fatty acids improve large artery endothelium-dependent dilation in subjects with hypercholesterolemia without affecting endothelium-independent dilation. PMID- 10676669 TI - Effects of oral L-arginine on endothelium-dependent vasodilation and markers of inflammation in healthy postmenopausal women. AB - OBJECTIVES: We examined whether oral administration of L-arginine, the substrate for nitric oxide (NO) synthesis, increases NO bioactivity in healthy postmenopausal women. BACKGROUND: Nitric oxide may protect arteries against atherosclerosis, as suggested by experimental studies in animals. Estrogen therapy, which has been shown to increase NO bioactivity in the vasculature of healthy postmenopausal women, is not acceptable for long-term use by many women. METHODS: In a randomized, double-blind, crossover study, 10 postmenopausal women without additional risk factors for atherosclerosis received L-arginine 9 g or placebo daily for one month, with treatment periods separated by one month. Nitric oxide levels in serum (as an index of endothelial NO release), brachial artery endothelium-dependent dilator responses to hyperemia by ultrasonography (as an index of vascular NO bioactivity) and markers of inflammation in blood that are inhibited by NO in cell culture experiments were measured at the end of each treatment period. RESULTS: L-arginine levels in plasma were increased in all women during L-arginine treatment compared with placebo (136.8 +/- 63.1 vs. 75.2 +/- 16.2 micromol/liter, p = 0.009). However, there was no change in serum nitrogen oxide levels (42.1 +/- 24.5 vs. 39.1 +/- 16.6 micromol/liter, p = 0.61), nor was there an effect of L-arginine on flow-mediated dilation during hyperemia (3.8 +/- 3.0% vs. 4.9 +/- 4.8%, p = 0.53) compared with placebo. Our study had sufficient power (beta = 0.80) to detect a true absolute treatment difference in flow-mediated brachial artery dilation of 1.7% or larger as statistically significant at alpha = 0.05. There was no effect of L-arginine on serum levels of soluble cell adhesion molecules compared with placebo: E-selectin (50.6 +/- 14.8 vs. 52.1 +/- 17.0 ng/ml, p = 0.45), intercellular adhesion molecule-1 (230 +/- 51 vs. 230 +/- 52 ng/ml, p = 0.97) and vascular cell adhesion molecule-1 (456 +/- 62 vs. 469 +/- 91 ng/ml, p = 0.53). CONCLUSIONS: Oral administration of L-arginine may not augment endothelial NO synthesis and release in postmenopausal women and is thus unlikely to be of general benefit to healthy postmenopausal women in protection from the development of atherosclerosis. PMID- 10676670 TI - Effects of vitamin E on chronic and acute endothelial dysfunction in smokers. AB - OBJECTIVES: The aims of this study were to determine whether chronic or acute impairment of flow mediated vasodilation (FMD) in the brachial artery of smokers can be restored or preserved by the antioxidant vitamin E. BACKGROUND: Transient impairment of endothelial function after heavy cigarette smoking and chronic endothelial dysfunction in smokers result at least in part from increased oxidative stress. METHODS: We studied 22 healthy male smokers (mean +/- SD, 23 +/ 9 cigarettes per day) randomly assigned to receive either 600 IU vitamin E per day (n = 11, age 28 +/- 6 years) or placebo (n = 11, age 27 +/- 6 years) for four weeks and 11 age-matched healthy male nonsmokers. Flow mediated vasodilation and endothelium-independent, nitroglycerin-induced dilation were assessed in the brachial artery using high resolution ultrasound (7.5 MHz) at baseline and after therapy. Subjects stopped smoking 2 h before the ultrasound examinations. At the end of the treatment period, a third scan was obtained 20 min after smoking a cigarette (0.6 mg nicotine, 7 mg tar) to estimate transient impairment of FMD. RESULTS: Flow mediated vasodilation at baseline was abnormal in the vitamin E (5.3 +/- 3.8, p < 0.01) and in the placebo group (6.4 +/- 3.5, p < 0.05) compared with nonsmoking controls (11.6 +/- 4.7). Using a two-way repeated measures analysis of variance (ANOVA) to examine the effects of vitamin E on FMD, we found no effect for the grouping factor (p = 0.5834) in the ANOVA over time but a highly significant difference with respect to time (p = 0.0065). The interaction of the time factor and the grouping factor also proved to be significant (p = 0.0318). Flow mediated vasodilation values remained similar after treatment for four weeks in both groups but declined faster after smoking a cigarette in subjects taking placebo compared with those receiving vitamin E (p values from successive differences for the time/group factor: 0.0001/0.0017). The transient attenuation of FMD (calculated as the percent change in FMD) was related to the improvement of the antioxidant status, estimated as percent changes in thiobarbituric acid-reactive substances (r = -0.67, p = 0.0024). Nitroglycerin induced dilation did not differ between study groups at baseline or after therapy. CONCLUSIONS: These results demonstrate that oral supplementation of vitamin E can attenuate transient impairment of endothelial function after heavy smoking due to an improvement of the oxidative status but cannot restore chronic endothelial dysfunction within four weeks in healthy male smokers. PMID- 10676671 TI - A comparison of angiotensin-converting enzyme inhibitors, calcium antagonists, beta-blockers and diuretic agents on reactive hyperemia in patients with essential hypertension: a multicenter study. AB - OBJECTIVES: The purpose of this study was to compare the effect of different antihypertensive agents, calcium antagonists, angiotensin-converting enzyme (ACE) inhibitors, beta-blockers and diuretic agents on endothelial function. BACKGROUND: Endothelial dysfunction is a component of essential hypertension, and various antihypertensive drugs may be able to restore normal function. METHODS: Forearm blood flow (FBF) was measured in 296 patients with essential hypertension, including 46 untreated subjects using strain-gauge plethysmography during reactive hyperemia and after sublingual administration of nitroglycerin (NTG). Forty-seven normotensive subjects were similarly evaluated as control subjects. RESULTS: The FBF during reactive hyperemia in the 296 hypertensive patients was significantly less than that in age-matched normotensive subjects. The increase in FBF after administration of sublingual NTG was similar in both groups. Systolic and diastolic blood pressures and forearm vascular resistance were greater in the untreated group than in the four treated groups and did not differ with respect to the antihypertensive agent used. The maximal FBF response from reactive hyperemia was significantly greater in the ACE inhibitor-treated group than in the group treated with calcium antagonists, beta-blockers, diuretic agents, or nothing (40.5 +/- 5.2 vs. 32.9 +/- 5.8, 34.0 +/- 5.6, 32.1 +/- 5.9, and 31.9 +/- 5.8 ml/min per 100 ml tissue, p < 0.05, respectively). Reactive hyperemia was similar in the calcium antagonist, beta-blocker, diuretic and untreated groups, and changes in FBF after sublingual NTG administration were similar in all groups. The infusion of NG-monomethyl-L-arginine, a nitric oxide (NO) synthase inhibitor, abolished the enhancement of reactive hyperemia in hypertensive patients treated with ACE inhibitors. CONCLUSIONS: These findings suggest that ACE inhibitors augment reactive hyperemia, an index of endothelium dependent vasorelaxation, in patients with essential hypertension. This augmentation may be due to increases in NO. PMID- 10676672 TI - Endothelial vasodilator function is related to low-density lipoprotein particle size and low-density lipoprotein vitamin E content in type 1 diabetes. AB - OBJECTIVES: We sought to determine whether endothelial vasodilator function (EVF) in patients with type 1 diabetes was related to low-density lipoprotein (LDL) particle size (LDLPS), LDL vitamin E content (LDLVE) or the susceptibility of LDL to oxidation (OxLDL). BACKGROUND: Impaired EVF is an early feature of diabetic vascular disease and may be related to oxidant stress. Although small, dense LDL and oxidized LDL are features of type 2 diabetes and predict the development of coronary artery disease, their role in type 1 diabetes is less clear. METHODS: Endothelium-dependent vasodilation was assessed in the brachial artery (flow mediated vasodilation [FMD]) and in the forearm resistance circulation using venous occlusion plethysmography in response to graded doses of intrabrachial acetylcholine (ACh). Thirty-seven patients with type 1 diabetes mellitus (DM) and 45 matched controls underwent flow-mediated dilation, while a subset of 19 DM and 20 controls underwent plethysmography. RESULTS: Total, LDL and high-density lipoprotein cholesterol or triglycerides were not different in DM compared with controls, but LDLPS was smaller (25.6 +/- 0.06 vs. 26.1 +/- 0.1 nm, p < 0.05) and LDLVE was reduced (2.0 +/- 0.25 vs. 2.6 +/- 0.18 micromol/mmol LDL, p < 0.05). Oxidative susceptibility of LDL was not different. Flow-mediated vasodilation was impaired in DM compared with controls (3.6 +/- 0.6% vs. 7.1 +/- 0.5%, p < 0.005), as was the vasodilator response to ACh (p < 0.05). Flow-mediated vasodilation was directly related to LDLPS and LDLVE in both the entire study cohort and DM alone (p < 0.05), but not to other parameters of the standard lipid profile. Similarly, endothelium-dependent vasodilation in the resistance circulation was directly related to LDLPS and LDLVE, but not to OxLDL. CONCLUSION: These results suggest, but do not prove, that LDL particle size and LDL vitamin E may be determinants of conduit and resistance vessel endothelial vasodilator function in type 1 diabetes. Further work will be required to prove cause and effect. PMID- 10676673 TI - Blood glucose and platelet-dependent thrombosis in patients with coronary artery disease. AB - OBJECTIVES: To investigate the influence of blood glucose on platelet-dependent thrombosis (PDT). BACKGROUND: Elevated blood glucose is a predictor of adverse cardiovascular risk independent of a diagnosis of diabetes, possibly due to adverse effects promoting thrombosis. The effects of blood glucose on PDT have not been characterized. METHODS: An ex vivo extracorporeal perfusion protocol was used to measure PDT in 42 patients with stable coronary artery disease (CAD). The Badimon chamber was perfused with unanticoagulated venous blood and PDT evaluated using computerized morphometry. Whole blood impedance aggregometry and flow cytometry evaluated platelet aggregation and P-selectin expression, respectively. RESULTS: Using a multivariate stepwise regression model, blood glucose was the best independent predictor of PDT (R2 = 0.19, p < 0.008), followed by apolipoprotein B (R2 = 0.18, p = 0.002) and intracellular magnesium levels (R2 = 0.12, p = 0.02). Platelet-dependent thrombosis was significantly greater in patients with blood glucose >, compared with <, the median value of 4.9 mmol/l (159 +/- 141 vs. 67 +/- 69 microm2/mm, p < 0.01). Neither platelet aggregation nor P-selectin expression was significantly different between the two groups. Insulin levels correlated with blood glucose (r = 0.56, p = 0.0003), but were not independently associated with either PDT, platelet aggregation or P-selectin expression. A two-way analysis of variance demonstrated an interaction between insulin (>126 pmol/l) and blood glucose (>4.9 mmol/l) in modulating PDT (F [1,38] = 8.5, p < 0.006). CONCLUSIONS: Blood glucose is an independent predictor of PDT in stable CAD patients. The relationship is evident even in the range of blood glucose levels considered normal, indicating that the risk associated with blood glucose may be continuous and graded. These findings suggest that the increased CAD risk associated with elevated blood glucose may be, in part, related to enhanced platelet-mediated thrombogenesis. PMID- 10676674 TI - Snoring and risk of cardiovascular disease in women. AB - OBJECTIVES: To examine prospectively the association between snoring and incidence of cardiovascular disease (CVD) in women. BACKGROUND: Whether snoring increases risk of CVD remains unclear; most previous studies have been small, not prospective and limited to men. METHODS: Seventy-one thousand seven hundred seventy-nine female nurses 40 through 65 years of age and without previously diagnosed CVD or cancer at baseline in 1986 were followed up for eight years. Frequency of snoring was assessed using mailed questionnaires at baseline. RESULTS: During eight years of follow-up, we documented 1,042 incident cases of major CVD events (644 coronary heart disease [CHD] and 398 stroke). Compared with nonsnorers, the age-adjusted relative risks (RRs) of CVD were 1.46 (95% confidence interval 1.23 to 1.74) for occasional snorers and 2.02 (1.62 to 2.53) for regular snorers. The age-adjusted RRs of CHD were 1.43 (1.15 to 1.77) for occasional snorers and 2.18 (1.65 to 2.87) for regular snorers. The age-adjusted RRs of stroke were 1.60 (1.21 to 2.12) and 1.88 (1.29 to 2.74), respectively. After further adjustment for smoking, body mass index (BMI) and other covariates, the positive association between snoring and CVD was attenuated but remained statistically significant (RRs of CVD were 1.20 [1.01 to 1.43] for occasional snorers and 1.33 [1.06-1.67] for regular snorers. CONCLUSIONS: These data suggested that snoring is associated with a modest but significantly increased risk of CVD in women, independent of age, smoking, BMI and other cardiovascular risk factors. While further study is needed to elucidate the biological mechanism underlying this association, snoring may help clinicians identify individuals at higher risk for CVD. PMID- 10676675 TI - Lack of association of lipoprotein(a) levels with coronary calcium deposits in asymptomatic postmenopausal women. AB - OBJECTIVES: This study sought to determine the relationship of lipoprotein(a) (Lp(a)) and other cardiac risk factors to coronary atherosclerosis as measured by calcification of coronary arteries in asymptomatic postmenopausal women. BACKGROUND: Lipoprotein(a) is considered a risk factor for coronary heart disease. Coronary calcium deposition is believed to be a useful noninvasive marker of coronary atherosclerosis in women. However, to our knowledge, there are no reports of the relationship of Lp(a) to coronary calcium in postmenopausal women. METHODS: In 178 asymptomatic postmenopausal women (64 +/- 8 years), we measured Lp(a) and other cardiac risk factors: age, hypertension, diabetes, low density lipoprotein cholesterol, smoking status, body mass index, physical activity level and duration of hormone replacement therapy. Electron-beam computed tomography was done to measure coronary calcium (calcium score). We analyzed the relationship between calcium score and cardiac risk factors using multivariate analysis. RESULTS: Although calcium score correlated with traditional risk factors of age, diabetes, hypertension and smoking, it did not correlate with Lp(a) in the asymptomatic postmenopausal women. Similar multivariate analyses were done in the subjects age >60 years and in the subjects with significant coronary calcium deposit (calcium score > or =50). These analyses also have failed to show an association of levels of Lp(a) with coronary calcium deposits. CONCLUSIONS: We conclude that in asymptomatic postmenopausal women, Lp(a) levels do not correlate with coronary atherosclerosis as measured by coronary calcium deposits. PMID- 10676676 TI - Garlic powder, effect on plasma lipids, postprandial lipemia, low-density lipoprotein particle size, high-density lipoprotein subclass distribution and lipoprotein(a). AB - OBJECTIVES: To test the hypothesis that a garlic supplement alters plasma lipoproteins, postprandial lipemia, low-density lipoprotein (LDL) size and high density lipoprotein (HDL) subclass distribution differently in 50 moderately hypercholesterolemic subjects classified as LDL subclass pattern A or B. BACKGROUND: Garlic has been variably reported to reduce or not affect plasma cholesterol values. Low-density lipoprotein pattern B is a common inherited disorder of lipoprotein metabolism that has been shown to have a significantly greater response to several lipid lowering treatments including low fat diet when compared with LDL pattern A individuals. METHODS: A double blind, randomized, placebo controlled trial in an outpatient lipid research clinic was performed and included fifty moderately hypercholesterolemic subjects (mean LDL cholesterol = 166 +/- 22 mg/dl) classified as LDL subclass pattern A (predominantly large LDL, n = 22) or B (predominantly small LDL, n = 28). Following a two-month stabilization period, subjects were randomly assigned to a placebo or 300 mg three times a day of a standardized garlic tablet for three months. RESULTS: For all subjects, LDL pattern A and B subjects combined, garlic treatment for three months resulted in no significant change in total cholesterol, LDL cholesterol, HDL cholesterol, HDL subclass distribution, postprandial triglycerides, apolipoprotein B, lipoprotein (a) (Lp[a]), LDL peak particle diameter or LDL subclass distribution. There was no significant difference in response for the same parameters among subjects classified as LDL pattern A or B with the exception of significantly greater (p = 0.01) reduction in mean peak particle diameter in pattern A subjects treated with either garlic or placebo. There was no significant change in LDL subclass distribution. CONCLUSIONS: This investigation confirms that garlic therapy has no effect on major plasma lipoproteins and further, that it has no impact on HDL subclasses, Lp(a), apolipoprotein B, postprandial triglycerides or LDL subclass distribution. Garlic may have a greater effect on LDL particle diameter in LDL pattern A compared with pattern B subjects. This difference was not reflected in other plasma lipid measurements. PMID- 10676677 TI - Coronary microcirculatory vasoconstriction during ischemia in patients with unstable angina. AB - OBJECTIVE: To verify the behavior of coronary microvascular tone during spontaneous ischemia in patients with unstable angina (UA). BACKGROUND: In UA, the pathogenetic role of vasoconstriction is classically confined at the stenotic coronary segment. However, microcirculatory vasoconstriction has been also suggested by previous experimental and clinical studies. METHODS: The study included 10 patients with UA (recent worsening of anginal threshold and appearance of angina at rest) and single-vessel CAD. Blood flow velocity was monitored by a Doppler catheter in the diseased artery. Transstenotic pressure gradient was monitored by aortic and distal coronary pressure monitoring. Stenosis resistance was calculated as the ratio between pressure gradient and blood flow, microvascular resistance as the ratio between distal pressure and blood flow. Measurements were obtained at baseline, following intracoronary adenosine (2 mg) and during transient ischemia. Aortic and distal coronary pressures were also measured during balloon coronary occlusion. RESULTS: Adenosine did not affect stenosis resistance, while it decreased (p < 0.05) microvascular resistance to 52 +/- 22% of baseline. Angina and ischemic ST segment shift were associated with transient angiographic coronary occlusion in 7 of 10 patients; however, in no case was ischemia associated with interruption of flow. Despite markedly different flow values, distal coronary pressure was similar during adenosine and during spontaneous ischemia (48 +/- 15 vs. 46 +/- 20 mm Hg, respectively, NS). During ischemia, a marked increase in the resistance of both coronary stenosis and coronary microcirculation was observed (to 1,233% +/- 1,298% and 671% +/- 652% of baseline, respectively, p < 0.05). Distal coronary pressure was markedly reduced during balloon coronary occlusion (14 +/- 7 mm Hg, p < 0.05 vs. both adenosine and ischemia), suggesting the absence of significant collateral circulation. CONCLUSIONS: In patients with UA, transient myocardial ischemia is associated with vasoconstriction of both stenotic arterial segment and downstream microcirculation. PMID- 10676678 TI - Identification of severe coronary artery disease in patients with a single abnormal coronary territory on exercise thallium-201 imaging: the importance of clinical and exercise variables. AB - OBJECTIVES: The aim of this study was to determine which clinical, exercise and thallium variables can aid in the identification of three-vessel or left main coronary artery disease (3VLMD) in patients with one abnormal coronary territory (either a reversible or fixed defect) on exercise thallium testing and to test the prognostic value of these variables. BACKGROUND: Although the sensitivity of detection of coronary artery disease by thallium-201 imaging is high, the actual detection of 3VLMD by thallium tomographic images alone is not optimal. METHODS: A multivariate model for prediction of 3VLMD was developed from several clinical, exercise and thallium-201 variables in a training population of 264 patients who had one abnormal coronary artery territory on exercise thallium testing and had undergone coronary angiography. Using this model, patients were stratified into risk groups for prediction of 3VLMD. A separate validation cohort of 474 consecutive patients who were treated initially with medical therapy and who had one abnormal coronary territory were divided into identical risk groupings by the variables derived from the training population, and they were followed for a median of 7.0 years to evaluate the prognostic value of this model. RESULTS: The prevalence of 3VLMD was 26% in the training population despite one abnormal thallium coronary territory. Four clinical and exercise variables--diabetes, hypertension, magnitude of ST segment depression, and exercise rate-pressure product-were found to be independent predictors of 3VLMD. In the training population, the prevalence of 3VLMD in low-, intermediate- and high-risk groups was 15%, 22% and 51%, respectively. When the multivariate model was applied to the validation population, the eight-year overall survival rates in the low-, intermediate- and high-risk groups were 89%, 73% and 75%, respectively (p < 0.001). CONCLUSIONS: A substantial proportion of patients with one abnormal thallium coronary territory have 3VLMD with subsequent divergent outcomes based upon risk stratification by clinical and exercise variables. Consequently, the finding of only a single abnormal coronary territory by thallium-201 perfusion imaging does not necessarily confer a benign prognosis in the absence of consideration of nonimaging variables. PMID- 10676679 TI - Three minute, but not one minute, ischemia and nicorandil have a preconditioning effect in patients with coronary artery disease. AB - OBJECTIVES: This study focused on 1) the determination of the optimal preconditioning (PC) duration, and 2) the protective effect of nicorandil (NC), a hybrid nitrate with a KATP channel opening effect, during a percutaneous transluminal coronary angioplasty (PTCA) model in humans. BACKGROUND: The ischemic PC effect is induced in 180 s ischemia, but not in 120 s ischemia in rabbit hearts. However, the duration of ischemia that induces PC effect and the role of the KATP channel in the PC effect in humans are still unclear. METHODS: Forty-six patients with stable angina were randomly allocated to four groups: the duration of the first inflation as PC ischemia was 60 s in the PC60 group (n = 12), and 180 s in the PC180 group (n = 12). In the other groups, NC (80 microg/kg) was intravenously given for 1 min in the NC group (n = 12), and isosorbide dinitrate (ISDN) (40 microg/kg) was given in the ISDN group (n = 10). Five minutes after first inflation or drug administration, a second inflation was conducted for 120 s in each group. In the ECG, the lead with the largest shift in ST segment (deltaST max), and the sum of elevated ST levels in all leads (sigmaST) were determined. RESULTS: In the PC60 group, no significant difference was observed in either deltaST max or sigmaST between the first and second inflation. However, the second inflation in the PC180 group showed significantly lower levels of deltaST max and sigmaST compared with those of the first inflation. In the NC group, both deltaST max and sigmaST measured at 30 s and 60 s after balloon inflation were significantly lower than those of the first inflation in the PC60 and PC180 control groups. In the ISDN group, no significant difference was observed in deltaST max or sigmaST. CONCLUSION: In human PTCA models, a PC effect is observed in 180 s ischemia, but not in 60 s ischemia. A pharmacological PC effect is induced by NC, a KATP channel opener with a nitrate like effect but not ISDN. This suggests that the opening of KATP channels plays an important role in the protecting effect of NC. PMID- 10676680 TI - Persistent ST segment depression in precordial leads V5-V6 after Q-wave anterior wall myocardial infarction is associated with restrictive physiology of the left ventricle. AB - OBJECTIVES: To examine the relationship between the persistence of ST segment depression in leads V5-V6 after Q-wave anterior wall myocardial infarction (MI) and the filling pattern of the left ventricle (LV). BACKGROUND: Precordial ST segment depression predominantly in leads V5-V6 is associated with increased in hospital morbidity and mortality after acute myocardial ischemia, perhaps due to reduced diastolic distensibility of the LV. METHODS: We prospectively studied 19 patients after Q-wave anterior wall MI (>6 months). All patients underwent 12 lead ECG recording, symptom-limited treadmill exercise testing with single photon emission computed tomography thallium-201 imaging, transthoracic Doppler echocardiography, cardiac catheterization and measurement of circulating atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) levels. Patients were classified based on the presence of ST segment depression in leads V5-V6: Group I = ST segment depression <0.1 mV (n = 10); Group II = ST segment depression > or =0.1 mV (n = 9). RESULTS: Patients in Group II had greater LV end diastolic pressures (32.4 +/- 6.5 mm Hg vs. 14.8 +/- 6.1 mm Hg; p = 0.0001), higher plasma ANP (44.4 +/- 47.1 pg/ml vs. 10.7 +/- 14 pg/ml; p = 0.04) and BNP levels (89.4 +/- 62.7 pg/ml vs. 23.6 +/- 33.1 pg/ml; p = 0.01), greater left atrium area (20.6 +/- 3.1 cm2 vs. 17.8 +/- 2.4 cm2; p = 0.05), lower peak atrial (A), higher early (E) mitral inflow velocities, a higher E/A ratio and a lower deceleration time (167 +/- 44 ms vs. 220 +/- 40 ms; p = 0.05). Lung thallium uptake during exercise was more common in Group II (78% vs. 10%, p = 0.04). CONCLUSIONS: Persistent ST segment depression in leads V5-V6 in survivors of Q wave anterior wall MI is associated with increased LV filling pressure and a restrictive LV filling pattern. PMID- 10676681 TI - Serum amyloid A predicts early mortality in acute coronary syndromes: A TIMI 11A substudy. AB - OBJECTIVES: We evaluated the ability of serum amyloid A (SAA), alone and in combination with a rapid qualitative assay for cardiac-specific troponin T (cTnT), to predict 14-day mortality in patients with unstable angina or non-Q wave myocardial infarction (NQMI). BACKGROUND: Elevated C-reactive protein (CRP) has been associated with adverse outcomes in unstable coronary syndromes but data regarding its acute phase counterpart, SAA, are conflicting. METHODS: Serum amyloid A measurement and a rapid cTnT assay were performed on blood obtained at enrollment into Thrombolysis in Myocardial Infarction 11A, a dose-ranging trial of enoxaparin for unstable angina and NQMI. RESULTS: Serum amyloid A was higher in patients who died compared with survivors (6.28 vs. 0.75 mg/dL, p = 0.002). Among patients with a negative rapid cTnT, mortality was higher for those in the top quintile of SAA (6.1 vs. 0.7%, p = 0.003). Patients with both an early positive rapid cTnT (< or =10 min until assay positive) and SAA in the fifth quintile had the highest mortality followed by those with either markedly elevated SAA or an early positive rapid cTnT, while patients with both a negative rapid cTnT and SAA in quintiles 1-4 were at very low risk, (9.1 vs. 3.6 vs. 0.7%, p <0.002). CONCLUSIONS: Similar to CRP, baseline elevation of SAA identifies patients hospitalized with unstable angina and NQMI at higher risk for early mortality, even among those with a negative rapid assay for cTnT. These data support further investigation of inflammatory markers used alone and in combination with cardiac troponins for risk assessment in unstable coronary syndromes. PMID- 10676682 TI - Ratio of left ventricular peak E-wave velocity to flow propagation velocity assessed by color M-mode Doppler echocardiography in first myocardial infarction: prognostic and clinical implications. AB - OBJECTIVES: To determine the ability of the ratio of peak E-wave velocity to flow propagation velocity (E/Vp) measured with color M-mode Doppler echocardiography to predict in-hospital heart failure and cardiac mortality in an unselected consecutive population with first myocardial infarction (MI). BACKGROUND: Several experimental studies indicate color M-mode echocardiography to be a valuable tool in the evaluation of diastolic function, but data regarding the clinical value are lacking. METHODS: Echocardiography was performed within 24 h of arrival at the coronary care unit in 110 consecutive patients with first MI. Highest Killip class was determined during hospitalization. Patients were divided into groups according to E/Vp <1.5 and > or =1.5. RESULTS: During hospitalization 53 patients were in Killip class > or =II. In patients with E/Vp > or =1.5, Killip class was significantly higher compared with patients with E/Vp <1.5 (p < 0.0001). Multivariate logistic regression analysis identified E/Vp > or =1.5 to be the single best predictor of in-hospital clinical heart failure when compared with age, heart rate, E-wave deceleration time (Dt), left ventricular (LV) ejection fraction, wall motion index, enzymatic infarct size and Q-wave MI. At day 35 survival in patients with E/Vp <1.5 was 98%, while for patients with E/Vp > or =1.5, it was 58% (p < 0.0001). Cox proportional hazards model identified Dt <140 ms, E/Vp > or =1.5 and age to be independent predictors of cardiac death, with Dt < 140 ms being superior to age and E/Vp. CONCLUSIONS: In the acute phase of MI, E/Vp > or =1.5 measured with color M-mode echocardiography is a strong predictor of in-hospital heart failure. Furthermore, E/Vp is superior to systolic measurements in predicting 35 day survival although Dt <140 ms is the most powerful predictor of cardiac death. PMID- 10676683 TI - Treatment and outcome of myocardial infarction in hospitals with and without invasive capability. Investigators in the National Registry of Myocardial Infarction. AB - OBJECTIVES: We sought to determine the extent to which the capability of a hospital to perform invasive cardiovascular procedures influences treatment and outcome of patients admitted with acute myocardial infarction (AMI). BACKGROUND: Patients with AMI are usually transported to the closest hospital. However, relatively few hospitals have the capability for immediate coronary arteriography, percutaneous transluminal coronary angioplasty (PTCA) or coronary artery bypass graft surgery (CABG), should these interventions be needed. METHODS: The 1,506 hospitals participating in the National Registry of Myocardial Infarction 2 were classified according to their highest level of invasive capability: 1) none (noninvasive, 28.1%); 2) coronary arteriography (cath capable, 25.2%); 3) coronary angioplasty (PTCA-capable, 7.4%); and 4) bypass surgery (CABG-capable, 39.2%). Treatment and in-hospital outcomes were assessed for 305,812 patients admitted from June 1994 through October 1996. Follow-up through 90 days was ascertained in a subset of 30,402 patients enrolled simultaneously in both the National Registry of Myocardial Infarction (NRMI) 2 and the Cooperative Cardiovascular Project (CCP). RESULTS: The proportion of patients receiving initial reperfusion intervention was only slightly higher at the more invasive hospitals (noninvasive 32.5%, cath-capable 31.2%, PTCA-capable 32.9% and CABG-capable 35.9%, p < 0.001 by chi-square statistic). Among thrombolytic recipients, median door-to-drug time interval differed little among hospital types and ranged from 42 to 45 minutes. At cath-capable, PTCA-capable and CABG-capable hospitals, coronary arteriography was performed in 32.9%, 37.4% and 64.9%, respectively, and PTCA in 0.0%, 5.1% and 31.4%, both p < 0.001 by chi square statistic. The proportion of patients transferred out to other facilities was 51.0%, 42.2%, 39.9% and 4.4% (p < 0.0001) among noninvasive, cath-capable, PTCA-capable and CABG-capable hospitals, respectively. Among patients in the combined NRMI and CCP data set, mortality at 90 days postinfarction was similar among patients initially admitted to each of the four hospital types. CONCLUSIONS: Although patients with AMI admitted to hospitals without invasive cardiac facilities have a high likelihood of subsequent transfer to other facilities, their likelihood of receiving a reperfusion intervention at the first hospital, their door to thrombolytic drug intervals and their 90-day survival rates are similar to those of patients initially admitted to more invasively equipped hospitals. These data suggest that a policy of initial treatment of myocardial infarction at the closest medical facility is appropriate medical practice. PMID- 10676684 TI - Management of myocardial infarction: looking beyond efficacy. PMID- 10676685 TI - Greater late lumen loss after successful coronary balloon angioplasty in the proximal left anterior descending coronary artery is not explained by extent of vessel wall damage or plaque burden. AB - OBJECTIVES: We investigated whether the greater late lumen loss after coronary balloon angioplasty in the proximal left anterior descending artery (P-LAD) compared with that in other segments might be related to differences in vascular dimensions or morphology as determined by angiography and intravascular ultrasound imaging. BACKGROUND: The greater late lumen loss after angioplasty in the P-LAD that has been observed in several studies has not been explained. METHODS: We studied 178 patients and 194 coronary artery lesions by quantitative angiography and 30 MHz intravascular ultrasound imaging after successful balloon angioplasty. Vessel wall morphology was compared among three proximal and three nonproximal segments. Follow-up quantitative angiography for late lumen loss calculation was performed in 168 lesions. Multivariate analysis was used to determine predictors of late lumen loss. RESULTS: Absolute and relative late loss were significantly greater at the P-LAD compared with the pooled group of other segments (0.42 +/- 0.60 mm vs. 0.10 +/- 0.48 mm, p = 0.0008 and 0.14 +/- 0.24 vs. 0.03 +/- 0.17, p < 0.001). Also, a greater percentage of calcific lesions (65% vs. 44%, p = 0.034), a lower incidence of rupture (51% vs. 74%, p = 0.009) and a larger reference segment plaque area (5.4 +/- 2.2 mm2 vs. 4.7 +/- 1.9 mm2, p = 0.05) were found in the P-LAD. In multivariate analysis however, these variables were not predictive of late loss. CONCLUSIONS: Greater late lumen loss after coronary balloon angioplasty of the P-LAD is not explained by differences in atherosclerotic plaque burden or in vessel wall damage. PMID- 10676686 TI - Procedural results and intermediate clinical outcomes after multiple saphenous vein graft stenting. AB - OBJECTIVES: We evaluated the early and mid-term (18-month) clinical events in a consecutive series of patients undergoing a nonstaged multiple saphenous vein grafting (SVG) intervention with stents as compared with a single SVG stent procedure. BACKGROUND: Saphenous vein graft angioplasty has been limited by high rates of distal embolization, myocardial infarction, restenosis and late mortality. It is unknown whether stenting of multiple, different SVGs at the same setting is associated with higher risk. METHODS: We evaluated in-hospital and mid term clinical outcomes (death, Q wave myocardial infarction [MI] and repeat revascularization rates up to 18 months) in 70 consecutive patients treated with coronary stents in 2 (93% of patients) or 3 SVGs, as compared with 649 patients undergoing stenting of a single SVG between January 1, 1994 and December 31, 1997. RESULTS: Overall procedural success was obtained in 97% of patients with 2 or 3 SVGs and 97% of patients with a single SVG (p = 0.94). Procedural complications were also similar (2.8% for multiple SVGs vs. 2.7% for a single SVG, p = 0.94). There was a higher prevalence of periprocedural non-Q wave MI (28% vs. 16%, p = 0.009) in the multiple SVG group. During follow-up (18 months), target lesion revascularization was 11% in multiple SVG and 15% in single SVG interventions (p = 0.19), and repeat revascularization (calculated per treated patient) was also similar for both groups (19% vs. 18%, p = 0.94). There was no difference in death (5.6% vs. 5.3%, p = 0.92) and Q wave MI rate (4.3% vs. 2.9%, p = 0.55) after the multiple SVG intervention. Overall cardiac event-free survival was similar for both groups (62% vs. 60%, p = 0.75). The study was powered to detect a clinically meaningful difference of 10% in mortality; smaller differences could not be evaluated on the basis of this sample size. CONCLUSIONS: Simultaneous stenting of multiple SVGs in carefully selected patients has similar in-hospital procedural success and major complications rates, as well as mid-term (18-month) clinical outcomes, as compared with single SVG stenting. Thus, multiple SVG interventions using stents may be a viable revascularization strategy for carefully selected patients and suitable lesions in multiple SVG disease. PMID- 10676687 TI - The effects of moxonidine, a novel imidazoline, on plasma norepinephrine in patients with congestive heart failure. Moxonidine Investigators. AB - OBJECTIVE: To evaluate the dose response relationship of moxonidine on plasma concentration of norepinephrine during acute and chronic administration in patients with congestive heart failure (CHF). BACKGROUND: Sympathetic activation is increased in heart failure. Moxonidine is an imidazoline ligand acting on the central nervous system (CNS) receptors to decrease sympathetic activation. METHODS: Ninety-seven patients with heart failure and New York Heart Association class II-III symptoms and ejection fraction <40% were randomized to placebo or one of three target doses of moxonidine, 0.1, 0.2 or 0.3 mg administered twice daily. An initial dose of moxonidine 0.1 mg twice a day (b.i.d.) was followed by weekly increments of 0.1 mg b.i.d. until target dose. The second and third study days occurred after four weeks (at target dose) and after 12 weeks, respectively. At each study day, repeated blood samples were drawn. RESULTS: There was a significant dose-related decrease of systolic blood pressure across all three study days. Heart rate decreased significantly on study day 3 in a dose-related manner. The acute 2 h decrease in plasma norepinephrine in response to all three doses of moxonidine was significantly different compared with placebo after four and 12 weeks. There was a significant linear relation between dose and plasma norepinephrine after four and 12 weeks in both 2 h peak and the time averaged effect (>8 h). The number of adverse events was similar in the moxonidine and placebo groups. CONCLUSIONS: The increased sympathetic activation in CHF can be reduced by moxonidine through CNS inhibition. PMID- 10676688 TI - Prediction of paroxysmal atrial fibrillation in patients with congestive heart failure: a prospective study. AB - OBJECTIVES: We sought to prospectively determine whether patients with congestive heart failure (CHF) at risk for paroxysmal atrial fibrillation (PAF) could be identified by clinical and study variables including the P-wave signal-averaged electrocardiogram (P-SAECG). BACKGROUND: Although it is important to assess the risk of developing PAF in patients with CHF, it still remains difficult to predict the PAF appearance in patients with CHF clinically. METHODS: The study group consisted of 75 patients in sinus rhythm without a history of PAF, whose left ventricular ejection fraction, as measured by radionuclide angiography, was <40%. These patients underwent P-SAECG, echocardiography and 24-h Holter monitoring; in addition, the plasma concentration of atrial natriuretic peptide (ANP) was measured at study entry. RESULTS: An abnormal P-SAECG was found at study entry in 29 of 75 patients. In the follow-up period of 21 +/- 9 months, the PAF attacks documented on the ECG significantly more frequently occurred in patients with (32%) rather than without an abnormal P-SAECG (2%) (p = 0.0002). The plasma ANP level was significantly higher in patients with rather than without PAF attacks (75 +/- 41 vs. 54 +/- 60 pg/ml, p = 0.01), although there were no significant differences in age, left atrial dimension or high grade atrial premature beats between the groups. The multivariate Cox analysis identified that the variables significantly associated with PAF development were an abnormal P-SAECG (hazard ratio 19.1, p = 0.0069) and elevated ANP level > or =60 pg/ml (hazard ratio 8.6, p = 0.018). CONCLUSIONS: An abnormal P-SAECG and elevated ANP level could be predictors of PAF development in patients with CHF. PMID- 10676689 TI - Mapping and ablation of ventricular tachycardia guided by virtual electrograms using a noncontact, computerized mapping system. AB - OBJECTIVES: The purpose of this study was to describe a computerized mapping system that utilizes a noncontact, 64 electrode balloon catheter to compute virtual electrograms simultaneously at 3,360 left ventricular (LV) sites and to assess the clinical utility of this system for mapping and ablating ventricular tachycardia (VT). BACKGROUND: Mapping VT in the electrophysiology laboratory conventionally is achieved by sequentially positioning an electrode catheter at multiple endocardial sites. METHODS: Fifteen patients with VT underwent 18 electrophysiology procedures using the noncontact, computerized mapping system. A 9F 64 electrode balloon catheter and a conventional 7F electrode catheter for mapping and ablation were positioned in the LV using a retrograde aortic approach. Using a boundary element inverse solution, 3,360 virtual endocardial electrograms were computed and used to derive isopotential maps. An incorporated locator system was used in conjunction with or instead of fluoroscopy to position the conventional electrode catheter. RESULTS: A total of 21 VTs, 12 of which were hemodynamically-tolerated and 9 of which were not, were mapped. Isolated diastolic potentials, presystolic areas, zones of slow conduction and exit sites during VT were identified using virtual electrograms and isopotential maps. Among 19 targeted VTs, radiofrequency ablation guided by the computerized mapping system and the locator signal was successful in 15. CONCLUSIONS: The computerized mapping system described in this study computes accurate isopotential maps that are a useful guide for ablation of hemodynamically stable or unstable VT. PMID- 10676690 TI - High dispersion of ventricular repolarization after an implantable defibrillator shock predicts induction of ventricular fibrillation as well as unsuccessful defibrillation. AB - OBJECTIVES: To test the hypothesis that post-shock dispersion of repolarization (PSDR) is higher in T wave shocks that induce ventricular fibrillation (VF) than in those that do not, as well as in implantable cardioverter defibrillator (ICD) defibrillation shocks which fail to terminate VF when compared with those that are successful. BACKGROUND: Ventricular fibrillation has been linked to the presence of dispersion of repolarization, which facilitates reentry. Most of the studies have been done in animals, and the mechanism underlying the generation and termination of VF in humans is speculative and remains to be determined. METHODS: Monophasic action potentials (MAPs) were recorded simultaneously from the right ventricular outflow tract (RVOT) and the right ventricular apex (RVA) in 27 patients who underwent implantation and testing of an ICD. T wave shocks were used to induce VF while the termination was attempted using internal defibrillator shocks. The post-shock repolarization time (PSRT) was measured in both the RVA and RVOT MAPs, and the difference between the two recordings was defined as the PSDR. The averages of PSDR were compared between the successful and unsuccessful inductions and terminations of VF. RESULTS: T wave shocks that induced VF generated a greater PSDR (93.4 +/- 85.1 ms) than the unsuccessful ones (45.1 +/- 55.9 ms, p < 0.001). On the other hand, shocks that failed to terminate VF were associated with a greater PSDR (59.9 +/- 41.2 ms) than shocks that terminated VF (21.1 +/- 20.1 ms), p < 0.001. CONCLUSIONS: A high PSDR following a T wave shock is associated with induction of VF; while following a defibrillating shock, it is associated with its failure and the continuation of VF. Conversely, a low PSDR is associated with failure of a T wave shock to induce VF and successful termination of VF by a defibrillating shock. PMID- 10676691 TI - Radiofrequency catheter ablation of supraventricular tachycardia substrates after mustard and senning operations for d-transposition of the great arteries. AB - OBJECTIVES: The purpose of this study was to determine the efficacy and risks of radiofrequency ablation of various forms of supraventricular tachycardia after Mustard and Senning operations for d-transposition of the great arteries. BACKGROUND: In this patient group, the reported success rate of catheter ablation of intraatrial reentry tachycardia is about 70% with a negligible complication rate. There are no reports of the use of radiofrequency ablation to treat other types of supraventricular tachycardia. METHODS: Standard diagnostic criteria were used to determine supraventricular tachycardia type. Appropriate sites for attempted ablation included 1) intraatrial reentry tachycardia: presence of concealed entrainment with a postpacing interval similar to tachycardia cycle length; 2) focal atrial tachycardia: a P-A interval < or =-20 ms; and 3) typical variety of atrioventricular (AV) node reentry tachycardia: combined electrographic and radiographic features. RESULTS: Nine Mustard and two Senning patients underwent 13 studies to successfully ablate all supraventricular tachycardia substrates in eight (73%) patients. Eight of eleven (73%) patients having intraatrial reentry tachycardia, 3/3 having typical AV node reentry tachycardia, and 2/2 having focal atrial reentry tachycardia were successfully ablated. Among five patients having intraatrial reentry tachycardia (IART) and not having ventriculoatrial (V-A) conduction, two suffered high-grade AV block when ablation of the systemic venous portion of the medial tricuspid valve/inferior vena cava isthmus was attempted. CONCLUSIONS: Radiofrequency catheter ablation can be effectively and safely performed for certain supraventricular tachycardia types in addition to intraatrial reentry. A novel catheter course is required for slow pathway modification. High-grade AV block is a potential risk of lesions placed in the systemic venous medial isthmus. PMID- 10676692 TI - Mechanism, localization and cure of atrial arrhythmias occurring after a new intraoperative endocardial radiofrequency ablation procedure for atrial fibrillation. AB - OBJECTIVES: The purpose of this study was to test a new pattern of radiofrequency ablation for atrial fibrillation (AFib) intended to optimize atrial activation, and to demonstrate the usefulness of catheter techniques for mapping and ablation of postoperative atrial arrhythmias. BACKGROUND: Linear radiofrequency lesions have been used to cure AFib, but the optimal pattern of lesions is unknown and postoperative tachyarrhythmias are common. METHODS: A radial pattern of linear radiofrequency lesions (Star) was made using an endocardial open surgical approach in 25 patients. Postoperative arrhythmias were induced and characterized during electrophysiological studies in 15 patients. RESULTS: The AFib was abolished in most patients (91%), but atrial flutter (AFlut) occurred in 96% of patients postoperatively. At postoperative electrophysiological studies, 37 flutter morphologies were studied in 15 patients (46% spontaneous, cycle length [CL] 223 +/- 25 ms). Seven mechanisms (lesions discontinuity, n = 6; focal mechanism, n = 1) of AFlut were characterized in six patients. In these cases, flutter was abolished using further catheter radiofrequency ablation. In the remaining cases, flutter was usually localized to an area involving the interatrial septum, but no critical isthmus was identified for ablation. After 16 +/-10 months, 15 patients (65%) were asymptomatic with (n = 3) or without (n = 12) antiarrhythmic medications. Eight (35%) patients had persistent arrhythmias. Postoperative atrial electrical activation was near physiological. CONCLUSIONS: The AFib maybe abolished using a radial pattern of linear endocardial radiofrequency lesions, but postoperative AFlut is common even when lesions are made under optimal conditions. Endocardial mapping techniques can be used to characterize the flutter mechanisms, thus enabling subsequent successful catheter ablation. PMID- 10676693 TI - Radiofrequency catheter ablation of inappropriate sinus tachycardia guided by activation mapping. AB - OBJECTIVE: The purpose of this study was to evaluate the value of activation mapping for radiofrequency modification of the sinus node and the long-term success rate of the procedure in a series of patients with inappropriate sinus tachycardia. BACKGROUND: The results of radiofrequency ablation of inappropriate sinus tachycardia have been reported in only a small number of patients. METHODS: The subjects of this study were 29 consecutive drug-refractory patients who underwent catheter ablation of inappropriate sinus tachycardia. Target sites were selected by activation mapping during sinus tachycardia. RESULTS: The ablation procedure was successful acutely in reducing the baseline sinus rate to <90/min and the sinus rate during isoproterenol infusion by >20% in 22 of 29 patients (76%). In 13 of 22 patients (59%) with a successful acute outcome, successive applications of radiofrequency energy at the site of earliest endocardial activation resulted in a cranial-caudal migration of earliest endocardial activation from the high lateral right atrium, along with a step-wise reduction in heart rate. In the other nine patients (41%) with a successful acute outcome, the reduction in sinus rate occurred abruptly, unaccompanied by migration of the site of earliest activation. Symptoms due to inappropriate sinus tachycardia recurred at a mean of 4.4+/-; 3 months after the ablation procedure in 6 of 22 patients (27%). After additional procedures in three patients, symptoms of inappropriate sinus tachycardia ultimately were successfully eliminated over the long-term in 19 of 29 patients (66%). CONCLUSIONS: In conclusion, radiofrequency ablation is at best only modestly effective for managing patients with inappropriate sinus tachycardia. The two different responses of heart rate to radiofrequency ablation may reflect differences in the number and/or multicentricity of subsidiary sites of impulse generation within the sinus node and/or atrium in patients with inappropriate sinus tachycardia. PMID- 10676694 TI - Efficacy and safety of catheter ablation in octogenarians. AB - OBJECTIVES: To determine whether catheter ablation is safe and effective in patients over the age of 80. BACKGROUND: There is a tendency to withhold invasive therapy in the elderly until it has been proven safe and effective. METHODS: Over a two-year period from February 1, 1996 to February 1, 1998, 695 consecutive patients underwent 744 catheter ablation procedures of supraventricular and ventricular arrhythmias. These patients were divided into three groups based on age: > or =80 years, 60 to 79 years and <60 years. Acute ablation success, using standard criteria and complication rates for these three groups were determined. RESULTS: There were 37 patients > or =80 years, 275 patients 60 to 79 years and 383 patients <60 years old. The overall acute ablation success rate for the entire group was 95% with no difference in rates among the three groups (97%, > or =80 years; 94%, 60-79 years; 95%, <60 years). The percentage of patients undergoing His bundle ablation was greatest in the > or =80-year-old group (43% vs. 19% vs. 2%, p < 0.01), and the percentage of patients undergoing accessory pathway ablation was greatest in the <60-year-old patients (0% vs. 4% vs. 25%, p < 0.01). The overall complication rate for the entire group was 2.6%, and there was only one major/life-threatening complication. There was no difference in complication rates among the groups (0%, > or =80 years; 2.2%, 60 to 79 years; 3.1%, <60 years). Based on the sample size, the 95% confidence interval is 0% to 7.8% for an adverse event in the octogenarian. CONCLUSIONS: Catheter ablative therapy for the arrhythmias attempted in the very elderly appears to be effective with low risk. Ablation results appear to be comparable with those noted in younger patients. PMID- 10676695 TI - Pulmonary blood flow alters nitric oxide production in patients undergoing device closure of atrial septal defects. AB - OBJECTIVE: To determine the effect of pulmonary blood flow (Qp) on nitric oxide (NO) production in patients with increased Qp due to an atrial septal defect (ASD). BACKGROUND: Alterations in pulmonary vascular NO production have been implicated in the development of pulmonary hypertension secondary to increased Qp. In vitro, acute changes in flow or shear stress alter NO production. However, the effect of Qp on lung NO production in vivo is unclear. METHODS: Nineteen patients (2.4-61 years of age, median 17) with secundum ASD undergoing device closure were studied. Before, and 30 min after ASD closure, exhaled NO and plasma nitrate concentration were measured by chemiluminescence (NOA 280, Sievers, Boulder, Colorado). RESULTS: Before ASD closure, all patients had increased Qp (Qp: systemic blood flow [Qs] of 2.0 +/- 0.7) and normal mean pulmonary arterial pressure (13.4 +/- 3.1 mm Hg). Atrial septal defect device closure decreased Qp from 6.0 +/- 2.5 to 3.6 +/- 1.3 L/min/m2 (p < 0.05). Mean pulmonary arterial pressure was unchanged. Associated with the decrease in Qp, both exhaled NO ( 22.1%, p < 0.05) and plasma nitrate concentrations (-17.9%, p < 0.05) decreased. CONCLUSIONS: These data represent the first demonstration that acute changes in Qp alter pulmonary NO production in vivo in humans. Exhaled NO determinations may provide a noninvasive assessment of pulmonary vascular NO production in patients with congenital heart disease. Potential correlations between exhaled NO, pulmonary vascular reactivity and pulmonary hypertension warrant further study. PMID- 10676696 TI - Pulmonary atresia with intact ventricular septum percutaneous radiofrequency assisted valvotomy and balloon dilation versus surgical valvotomy and Blalock Taussig shunt. AB - OBJECTIVE: We compared the result of radiofrequency (RF)-assisted valvotomy and balloon dilation with closed surgical valvotomy and Blalock Taussig (BT) shunt as primary treatment in selected patients with pulmonary atresia and intact ventricular septum (PA-IVS). BACKGROUND: Patients with PA-IVS who have mild to moderate hypoplasia of the right ventricle (RV) and patent infundibulum have the greatest potential for complete biventricular circulation. The use of RF or laser wires to perforate the atretic valve followed by balloon dilation provides an alternative to surgery. METHODS: Between May 1990 and March 1998, 33 selected patients underwent either percutaneous RF valvotomy and balloon dilation (group 1, n = 21; two crossed over to group 2) or surgical valvotomy with concomitant BT shunt (group 2, n = 14). Second RV decompression by balloon dilation or right ventricular outflow tract (RVOT) reconstruction were performed if necessary. Patients who remained cyanosed were subjected to transcatheter trial closure of the interatrial communication. Partial biventricular repair was offered to those with inadequate growth of the RV. RESULTS: The primary procedure was successful in 19 patients in group 1. There was one in-hospital death and two late deaths. Of the remaining 16 survivors, 12 achieved complete biventricular circulation, 7 of whom required no further interventions. Two patients required repeat balloon dilation, 1 RVOT reconstruction and 2 transcatheter closure of interatrial communication. Two patients underwent partial biventricular repair. In group 2, there were 3 in-hospital deaths after the primary procedure and 1 patient died four months later. All survivors (n = 10) required a second RV decompression, 8 by balloon dilation and 2 by RVOT reconstruction, after which, two patients died. Of the final 8 survivors, 7 achieved complete biventricular circulation, 5 after coil occlusion of the BT shunt and 2 after closure of interatrial communication. CONCLUSIONS: Radiofrequency valvotomy and balloon dilation is more efficacious and safe compared with closed pulmonary valvotomy and BT shunt in selected patients with PA-IVS. PMID- 10676697 TI - Importance of imaging method over imaging modality in noninvasive determination of left ventricular volumes and ejection fraction: assessment by two- and three dimensional echocardiography and magnetic resonance imaging. AB - OBJECTIVES: This study sought to determine the concordance between biplane and volumetric echocardiography and magnetic resonance imaging (MRI) strategies and their impact on the classification of patients according to left ventricular (LV) ejection fraction (EF) (LVEF). BACKGROUND: Transthoracic echocardiography and MRI are noninvasive imaging modalities well suited for serial evaluation of LV volume and LVEF. Despite the accuracy and reproducibility of volumetric methods, quantitative biplane methods are commonly used, as they minimize both scanning and analysis times. METHODS: Thirty-five adult subjects, including 25 patients with dilated cardiomyopathies, were evaluated by biplane and volumetric (cardiac short-axis stack) cine MRI and by biplane and volumetric (three-dimensional) transthoracic echocardiography. Left ventricular volume, LVEF and LV function categories (LVEF > or =55%, >35% to <55% and < or =35%) were then determined. RESULTS: Biplane echocardiography underestimated LV volume with respect to the other three strategies (p < 0.01). There were no significant differences (p > 0.05) between any of the strategies for quantitative LVEF. Volumetric MRI and volumetric echocardiography differed by a single functional category for 2 patients (8%). Six to 11 patients (24% to 44%) differed when comparing biplane and volumetric methods. Ten patients (40%) changed their functional status when biplane MRI and biplane echocardiography were compared; this comparison also revealed the greatest mean absolute difference in estimates of EF for those subjects whose EF functional category had changed. CONCLUSIONS: Volumetric MRI and volumetric echocardiographic measures of LV volume and LVEF agree well and give similar results when used to stratify patients with dilated cardiomyopathy according to systolic function. Agreement is poor between biplane and volumetric methods and worse between biplane methods, which assigned 40% of patients to different categories according to LVEF. The choice of imaging method (volumetric or biplane) has a greater impact on the results than does the choice of imaging modality (echocardiography or MRI) when measuring LV volume and systolic function. PMID- 10676698 TI - Contrast echocardiography clarifies uninterpretable wall motion in intensive care unit patients. AB - OBJECTIVES: The study examined the value of contrast echocardiography in the assessment of left ventricular (LV) wall motion in intensive care unit (ICU) patients. BACKGROUND: Echocardiograms done in the ICU are often suboptimal. The most common indication is the evaluation of LV wall motion and ejection fraction (EF). METHODS: Transthoracic echocardiograms were done in 70 unselected ICU patients. Wall motion was evaluated on standard echocardiography (SE), harmonic echocardiography (HE), and after intravenous (IV) contrast echocardiography (CE) using a score for each of 16 segments. A confidence score was also given for each segment with each technique (unable to judge; not sure; sure). The EF was estimated visually for each technique, and a confidence score was applied to the EF. RESULTS: Uninterpretable wall motion was present in 5.4 segments/patient on SE, 4.4 on HE (p = 0.2), and 1.1 on CE (p < 0.0001). An average of 7.8 segments were read with surety on SE, 9.2 on HE (p = 0.1), and 13.7 on CE (p < 0.0001). Ejection fraction was uninterpretable in 23% on SE, 13% on HE (p = 0.14), and 0% on CE (p = 0.002 vs. HE; p < 0.0001 vs. SE). The EF was read with surety in 56% of patients on SE, 62% on HE (p = 0.47), and 91% on CE (p < 0.0001). Thus, wall motion was seen with more confidence on CE. More importantly, the actual readings of segmental wall motion and EF significantly differed using CE. CONCLUSIONS: CE should be used in all ICU patients with suboptimal transthoracic echocardiograms. PMID- 10676699 TI - Improving the evaluation of left ventricular systolic function with intravenous perfluorocarbon ultrasound contrast: will suboptimal echocardiograms become an endangered species? PMID- 10676700 TI - Polymeric-based perivascular delivery of a nitric oxide donor inhibits intimal thickening after balloon denudation arterial injury: role of nuclear factor kappaB. AB - OBJECTIVES: To examine the effect of a polymeric-based periadventitial delivery of a nitric oxide (NO)-releasing diazeniumdiolate, spermine/NO (SPER/NO), on balloon injury-induced neointimal hyperplasia in rat ileofemoral arteries. BACKGROUND: Reduced local bioavailability and adverse side effects limit systemic administration of NO to modulate vascular response to injury. METHODS: A polylactic-polyglycolic acid polymeric matrix containing 2.5% SPER/NO (w/w) was applied around the injured arteries. Quantitative histomorphometry was performed at day 14, proliferating cell nuclear antigen (PCNA) immunohistochemistry at day 3 to assess effects on smooth muscle proliferation and electrophoretic mobility shift assay to evaluate effects on transcription factor, nuclear factor-kappaB (NF-kappaB). RESULTS: Treatment with SPER/NO reduced the intimal area (0.011 +/- 0.009 vs. 0.035 +/- 0.006 mm2 control, p < 0.01) and the intima to media ratio (0.089 +/- 0.062 vs. 0.330 +/- 0.057 control, p < 0.005). Spermine/nitric oxide produced a profound inhibition of PCNA-positive cells (>75%, p < 0.005) and significantly suppressed the injury-induced activation of NF-kappaB. Vascular cyclic guanosine monophosphate (cGMP) levels were elevated after treatment with the SPER/NO (0.28 +/- 0.03 vs. 0.17 +/- 0.02 pmol/mg tissue control, p < 0.01). The inhibitory effects on neointimal proliferation were localized to the site of application of SPER/NO and were not associated with any changes in platelet aggregation or bleeding time. Neither SPER nor polymer alone had any significant effects on any of the variables examined. CONCLUSIONS: Polymeric-based perivascular delivery of a NO donor produces a marked localized inhibition of neointimal proliferation in balloon-injured arteries. This phenomenon is associated with suppression of NF-kappaB activation and elevation of the vascular cGMP at the site of injury. PMID- 10676701 TI - Single-beat determination of preload recruitable stroke work relationship: derivation and evaluation in conscious dogs. AB - OBJECTIVES: To derive and evaluate a method of estimating the slope (Mw) of the preload recruitable stroke work (PRSW) relationship between left ventricular stroke work (SW) and end-diastolic volume (EDV) from a single beat. BACKGROUND: Mw is a load-insensitive index of contractile function, but its clinical application has been limited by the need to record multiple beats over a wide volume range. METHODS: Pressure-volume loops were recorded over a variable preload and afterload range by vena caval and aortic constrictions in 12 conscious dogs instrumented with epicardial dimension transducers and micromanometers. Single-beat Mw (SBMw) was determined as the ratio SW/(EDV-Vw), where the volume-axis intercept of the PRSW relationship (Vw)(EDV at zero SW) was estimated as k x EDVB + (k - 1)LVwall, k is the ratio of the epicardial shell volumes corresponding to Vw and baseline EDV (EDVB) and LVwall is wall volume. RESULTS: In the first six dogs, k was found to be essentially constant at 0.7, SBMw estimates were insensitive to wide preload variation, and the relationship between SBMw and multibeat Mw determined during caval and aortic constrictions did not differ significantly from the line of identity. When the same constant k value was applied to SBMw estimation in a different group of six dogs, SBMw did not differ significantly from multibeat Mw (83 +/- 12 erg x cm(-3) x 10(3) and 77 +/- 12 erg x cm(-3) x 10(3), respectively), neither changed significantly during aortic constriction and both increased significantly with calcium infusion (107 +/- 18 erg x cm(-3) x 10(3) and 95 +/- 19 erg x cm(-3) x 10(3), respectively, both p < 0.05). Single-beat Mw was less load-dependent, more reproducible and a more sensitive index of inotropic state than two previously described single-beat indexes, single-beat elastance and maximum power divided by EDV2. CONCLUSIONS: Mw can be determined accurately from a single, steady-state beat in the normal canine heart and is sensitive to inotropic alterations while being insensitive to wide variations in preload and afterload. Single-beat Mw estimation should facilitate noninvasive, load-independent assessment of contractile function. PMID- 10676702 TI - Nicorandil, a potent cardioprotective agent, acts by opening mitochondrial ATP dependent potassium channels. AB - OBJECTIVES: To determine the mechanism of cardioprotection afforded by nicorandil, an orally efficacious antianginal drug, we examined its effects on ATP-dependent potassium (K(ATP)) channels. BACKGROUND: Nicorandil can mimic ischemic preconditioning, while mitochondrial K(ATP) (mitoK(ATP)) channels rather than sarcolemmal K(ATP) (surfaceK(ATP)) channels have emerged as the likely effectors. METHODS: Flavoprotein fluorescence and membrane current in intact rabbit ventricular myocytes were measured simultaneously to assay mitoK(ATP) channel and surface K(ATP) channel activities, respectively. In a cell-pelleting model of ischemia, cells permeable to trypan blue were counted as killed by 60 and 120 min of ischemia. RESULTS: Nicorandil (100 micromol/liter) increased flavoprotein oxidation but not membrane current; a 10-fold higher concentration recruits both mitoK(ATP) and surfaceK(ATP) channels. Pooled dose-response data confirm that nicorandil concentrations as low as 10 micromol/liter turn on mitoK(ATP) channels, while surfaceK(ATP) current requires exposure to millimolar concentrations. Nicorandil blunted the rate of cell death in a pelleting model of ischemia; this cardioprotective effect was prevented by the mitoK(ATP) channel blocker 5-hydroxydecanoate but was unaffected by the surfaceK(ATP) channel blocker HMR1098. CONCLUSIONS: Nicorandil exerts a direct cardioprotective effect on heart muscle cells, an effect mediated by selective activation of mitoK(ATP) channels. PMID- 10676703 TI - Intracoronary basic fibroblast growth factor enhances myocardial collateral perfusion in dogs. AB - OBJECTIVE: In preparation for clinical trials of basic fibroblast growth factor (bFGF) to treat ischemic heart disease, we sought to identify a clinically feasible method of bFGF administration. BACKGROUND: Basic FGF has been shown to promote collateral development after experimentally induced coronary occlusion; however, methods of bFGF delivery that have been shown to be effective in previous investigations would not be practical for clinical use. METHODS: Four randomized, blinded, controlled investigations were conducted independently and sequentially in an established canine model. For all studies, dogs underwent operative placement of proximal left circumflex coronary artery ameroid constrictors. The four investigational regimens included: 1) bFGF by central venous bolus injection, 1,740 microg/day for one, two or seven days; 2) bFGF by intravenous infusion, 100 microg/kg body weight per day for seven days; 3) bFGF by pericardial instillation, 2,000 microg/day for 7 days; and 4) bFGF by intracoronary injection (Judkin's technique), 100 microg/kg per day for one or two days. Each substudy included a contemporaneous vehicle control group. Collateral perfusion (microspheres) was assessed during maximal coronary vasodilation during the first month after ameroid placement. RESULTS: Maximal collateral perfusion in dogs that received intracoronary bFGF for two days exceeded that of concurrent control dogs by 31% (p < 0.01). Perfusion was not increased in dogs that received single-dose intracoronary bFGF. Basic FGF administration by central venous bolus injection, intravenous infusion and pericardial injection failed to enhance collateral perfusion. CONCLUSIONS: Administration of bFGF by the intracoronary route, an intervention that is feasible in patients, augments collateral development in dogs. These data provide a rationale for clinical testing of intracoronary bFGF in ischemic heart disease. PMID- 10676704 TI - Survival with full neurologic recovery and no cerebral pathology after prolonged cardiopulmonary resuscitation with vasopressin in pigs. AB - OBJECTIVES: We sought to determine the effects of vasopressin and saline placebo in comparison with epinephrine on neurologic recovery and possible cerebral pathology in an established porcine model of prolonged cardiopulmonary resuscitation (CPR). BACKGROUND: It is unknown whether increased cerebral blood flow during CPR with vasopressin is beneficial with regard to neurologic recovery or detrimental owing to complications such as cerebral edema after return of spontaneous circulation. METHODS: After 4 min of cardiac arrest, followed by 3 min of basic life support CPR, 17 animals were randomly assigned to receive every 5 min either vasopressin (0.4, 0.4 and 0.8 U/kg; n = 6), epinephrine (45, 45 and 200 microg/kg; n = 6) or saline placebo (n = 5). The mean value +/- SEM of aortic diastolic pressure was significantly (p < 0.05) higher 90 s after each of three vasopressin versus epinephrine versus saline placebo injections (60 +/- 3 vs. 45 +/- 3 vs. 29 +/- 2 mm Hg; 49 +/- 5 vs. 27 +/- 3 vs. 23 +/- 1 mm Hg; and 50 +/- 6 vs. 21 +/- 3 vs. 16 +/- 3 mm Hg, respectively). After 22 min of cardiac arrest, including 18 min of CPR, defibrillation was attempted to achieve return of spontaneous circulation. RESULTS: All the pigs that received epinephrine and saline placebo died, whereas all pigs on vasopressin survived (p < 0.05). Neurologic evaluation 24 h after successful resuscitation revealed only an unsteady gait in all vasopressin-treated animals; after 96 h, magnetic resonance imaging revealed no cerebral pathology. CONCLUSIONS: During prolonged CPR, repeated vasopressin administration, but not epinephrine or saline placebo, ensured long-term survival with full neurologic recovery and no cerebral pathology in this porcine CPR model. PMID- 10676705 TI - Heart failure awareness week: February 14-21. PMID- 10676706 TI - President's page: back to the future: part III. PMID- 10676707 TI - Fluorescence in situ hybridization (FISH) maps chromosomal homologies between the dusky titi and squirrel monkey. AB - The Platyrrhini are one of the most karyologically derived groups of primates and the evolution of their karyotypes is far from understood. The identification of the origin and direction of chromosome rearrangements will contribute to a better understanding of New World monkey phylogeny, taxonomy, and evolution. We mapped homology and identified translocations in the chromosomes of the dusky titi monkey (Callicebus moloch, 2n = 50) and the squirrel monkey (Saimiri sciureus, 2n = 44) by fluorescence in situ hybridization (FISH) of human chromosome paints. The hybridization results established chromosomal homologies between these New World primates, humans, other primates, and more distantly related mammalian species and show that both species have highly rearranged karyotypes. The total number of hybridization signals was 37 in C. moloch and 40 in S. sciureus, which is in the range of most comparisons of human chromosomes with phylogenetically more distant species outside of the primate order. Parsimony analyses of outgroup painting patterns allowed us to propose an ancestral karyotype for New World monkeys consisting of 2n = 56 with homologs to the following human chromosomes or chromosome segments: 1b; 1c; 2a; 2b; 3a; 3b; 3/21; 4; 5; 6; 7; 8a; 8/18; 9; 10a; 10/16; 11; 12; 13; 14/15; 15a; 16a; 17; 19; 20; 22; X; Y. Associations 8/18 and 10/16 are derived ancestral associations for all Platyrrhini. A 2/16 association found in S. sciureus and C. moloch was also seen in Ateles geoffroyi and Cebus capucinus; a 5/7 association in S. sciureus was present in A. geoffroyi, C. capucinus, and Alouatta belzebul. Other associations seen in the dusky titi monkey or the squirrel monkey are probably automorphisms. Comparison with chromosome phylogenies based on R-banding [Dutrillaux et al., 1986] showed that there were many errors in assigning homology with human chromosomes. The chromosomal phylogeny of New World monkeys based on banding patterns is in need of revision using modern molecular methods. PMID- 10676708 TI - When will the stork arrive? Patterns of birth seasonality in neotropical primates. AB - We review and discuss the ultimate and proximate causes of birth seasonality in Neotropical primates and the seasonal patterns shown by each genus within this group. Our review of the literature shows that most New World monkey populations studied so far show some degree of birth seasonality. Photoperiod is the most important proximate cue used by populations living at relatively high latitudes to time their reproductive events, but almost nothing is known about the proximate factors used by those near the equator. The findings are consistent with the hypothesis that food availability is the most important ultimate cause of birth seasonality. Predation seems to promote birth synchrony in some species (e.g., squirrel monkeys). Multiple regression ANCOVA was used to estimate how the degree of birth seasonality is affected by ecological and life history variables. The ANCOVA model shows that three factors affect the degree of birth seasonality: diet, latitude, and body size. Folivores (howlers) are less seasonal than frugivores and insectivores. The degree of seasonality increases with latitude and shows a humped relationship with body size, peaking at 1.66 kg body mass. This last relationship was expected since small bodied species have to pay a cost (in terms of time lost) by being seasonal on a yearly basis, and large species are buffered against fluctuations in food availability due to their large body mass. To understand which of three alternative birth strategies is followed by each species (reduce energy stress during peak lactation, wean infants during peak food availability, or store reserves during peak energy availability), we compared the location of the birth peak in relation to the peak in food availability for those populations from which data were available. Most species conform to the typical pattern of births concentrated before the peak in food availability, allowing peak lactation (small-sized species) or weaning (capuchins) to take place before the start of the lean season. The pattern of births of the atelines is consistent with the weaning hypothesis. However, since they give birth during the lean season, this pattern is also consistent with an alternative strategy. PMID- 10676709 TI - Survival and reproduction in the first two years following a large-scale primate colony move and social reorganization. AB - (Macaca nemestrina) and baboon (Papio cynocephalus, Papio anubis, and hybrids) breeding colonies from the Primate Field Station (PFS) (Medical Lake, WA) to the Tulane Regional Primate Research Center (Covington, LA). Colony records on all 598 pigtailed macaques (Macaca nemestrina) and 157 baboons (P. c. anubis) shipped to the Tulane Primate Center from the PFS breeding colony were used for analysis of species, sex, age, origin, current status, and the number of animals born at Tulane and their status. To provide comparative statistics, colony records on all 1,002 macaques and 258 baboons alive on 1 January 1991 at the Field Station were retrieved in the same manner as the Tulane data. Overall survival rates of macaques in the months following the move (71.7%) were similar to those associated with the Arashiyama West colony move from Japan to Texas. In our colony, significantly lower survival following the move was seen only in older (10 years+) macaques, while survival in other age groups was slightly lower than in the comparison year of 1991 at the Primate Field Station. Captive-bred macaques exhibited higher survival than wild-caught animals. Infant survival at Tulane was not significantly different than in pre-move years. Baboons fared well in the move, with no significant differences in mortality or reproduction when compared with the 1991 Medical Lake baboon colony. PMID- 10676710 TI - Diurnal primate densities and biomass in the Kakamega Forest: an evaluation of census methods and a comparison with other forests. AB - Line-transect surveys were conducted at the Isecheno study site in the Kakamega Forest, western Kenya to estimate diurnal primate densities. The estimates from several different methods of analysis of census data were compared to "true" density values based on home range size and overlap for two species. The Whitesides method [Whitesides et al., 1988], which incorporates species-specific mean group spread into its formula for estimating transect width, provided the most accurate density estimates. The importance of including as many groups as possible when calculating density from home range size and overlap is demonstrated with long-term data from Colobus guereza and Cercopithecus mitis. Colobus guereza group density at Isecheno was much lower than that published from a recent brief study [von Hippel, 1996]. Cercopithecus mitis group density has fallen while overall population biomass appears to have remained stable over 20 years of study. Isecheno has the second highest diurnal primate biomass of the ten Guineo-Congolian rainforest sites for which biomass data are available, despite having the lowest primate species richness. Within the Guineo-Congolian rainforest system, primate biomass appears to vary to some extent between ecogeographic regions: two of three mid-elevation East African sites have high biomasses, two of two lowland West African sites have intermediate biomasses, and four of five lowland Central African sites have low biomasses. There is a strong positive correlation between total colobine biomass and total primate biomass at the ten Guineo-Congolian rainforest sites. PMID- 10676711 TI - Infanticide in a group of wild saddle-back tamarins, Saguinus fuscicollis. AB - An infanticide was observed in a group of wild saddle-back tamarins, Saguinus fuscicollis. The newborn singleton was killed by its mother after it had fallen from the carrier several times. This infanticide may represent a case of parental manipulation: the mother terminated investment in an offspring that probably had a low chance of survival. Also, stress associated with the simultaneous pregnancy of another adult female in the group may have played a role. PMID- 10676712 TI - Reconciliation in captive Guyanese squirrel monkeys (Saimiri sciureus). AB - The tendency for agonistic interaction to increase the probability of friendly interaction between social partners has been demonstrated across a range of Old World primates. While research on such post-conflict behavior proceeds into an hypothesis-testing phase, new comparative information must accumulate to provide full phylogenetic perspective on primate social behavior. Data from New World and prosimian primates are yet extremely limited. We studied captive squirrel monkeys (Saimiri sciureus) via post-conflict (PC) and matched control (MC) observations and analyzed results using both the PC-MC and time-rule methods. Former opponents maintaining affiliative relationships soon engaged in friendly interaction following large proportions of agonistic interactions, whereas non-affiliated individuals, including virtually all male-female pairs, reconciled conflicts rarely. Close-proximity approaching and huddling contact constituted the principal modes of post-conflict amicability. Agonistic interactions of relatively high intensity were most likely to be reconciled and most likely to be reconciled via physical contact. High vulnerability of Saimiri to predation may have favored this species' strong inclination to reconcile soon after agonistic interaction. Research on free-living populations of this and other primate species is needed to illuminate similarities and differences across taxa. PMID- 10676713 TI - Beyond "adequate dialysis". PMID- 10676714 TI - The role of angiotensin II and plasminogen activator inhibitor-1 in progressive glomerulosclerosis. AB - Regardless of the primary cause, progressive renal deterioration with sclerosis is a hallmark of many renal diseases. Several studies have shown the superiority of angiotensin-converting enzyme inhibitors compared with other antihypertensive agents in providing protection from progressive renal deterioration. Furthermore, animal studies have shown that angiotensin II antagonists in excess of antihypertensive doses can also ameliorate or reverse glomerulosclerosis, leading to the hypothesis that angiotensin II has nonhemodynamic effects that mediate the renoprotective effects shown in these investigations. Although historically angiotensin II has been associated with salt and fluid homeostasis, recent data show that angiotensin II induces cell growth and matrix accumulation in glomerular cells. Plasminogen activator inhibitor-1 has been shown to be the major inhibitor of tissue plasminogen activator and urokinase-like plasminogen activator, with potentially important effects not only on thrombosis/fibrinolysis, but also on matrix degradation because of the proteolytic actions of these substances. Angiotensin II has been shown to influence the actions of plasminogen activator inhibitor-1 and, consequently, its thrombotic and sclerotic effects. Various studies, both in vitro and in vivo, have shown that direct hemodynamic actions, modulation of endothelial injury, and growth factor actions also may be important in the development of sclerosis. These factors can be directly modulated by angiotensin II inhibition. Sclerosis may even be reversed when therapies augment matrix degradation processes, both by directly increasing proteolytic activity and by downregulating inhibitors of matrix degradation. These observations indicate that angiotensin II is important in fibrotic as well as thrombotic renal injuries that lead to progressive renal disease and also in the development of therapies such as specific angiotensin receptor antagonists to prevent or reverse these conditions. PMID- 10676715 TI - Fatal outcome after ingestion of star fruit (Averrhoa carambola) in uremic patients. AB - Clinical outcome of dialysis patients after eating star fruit (Averrhoa carambola) varies, but it may be fatal. In the past 10 years, 20 such patients were treated in our hospital when they developed clinical symptoms after eating the fruit or drinking star fruit juice. Their initial presentations included sudden-onset limb numbness, muscle weakness, intractable hiccups, consciousness disturbance of various degrees, and seizure. No other major events that might be responsible for these symptoms could be identified. Eight patients died, including one patient with a serum creatinine level of 6.4 mg/dL who had not yet begun dialysis. The clinical manifestations of the survivors were similar to those who died except for consciousness disturbance and seizure. Death occurred within 5 days despite emergent hemodialysis and intensive medical care. The survivors' symptoms usually became less severe after supportive treatment, and these patients subsequently recovered without obvious sequelae. The purpose of this article is to report that patients with renal failure who ingest star fruit may develop neurological symptoms and also run the risk for death in severe cases. Mortality may also occur in patients with chronic renal failure not yet undergoing dialysis. PMID- 10676716 TI - Early treatment with corticosteroids ameliorates proteinuria, proliferative lesions, and mesangial phenotypic modulation in adult diffuse proliferative IgA nephropathy. AB - Diffuse proliferative immunoglobulin A (IgA) nephropathy has the potential risk for end-stage renal disease. However, treatment of IgA nephropathy has not been well established. To determine whether early treatment with corticosteroids ameliorates the proliferative lesions of diffuse proliferative IgA nephropathy, we conducted a prospective, randomized, controlled trial. Inclusion criteria were as follows: duration of abnormal urinalysis results less than 36 months, proteinuria less than 1.5 g/d of protein, serum creatinine level less than 1.5 mg/dL, and mesangial cell proliferation or matrix accumulation involving more than 50% of glomeruli. Twenty-one patients were randomly assigned to two groups: the corticosteroid group and the antiplatelet group. After 1 year of treatment, repeated renal biopsy was performed in 19 patients. We evaluated glomerular filtration rate, blood pressure, proteinuria, and histological parameters, including light microscopic findings and staining of alpha-smooth muscle actin (alphaSMA), as a marker of myofibroblast-like cells and fibronectin EDA (EDA-FN) as an indicator of renal fibrosis. After 1 year of treatment, proteinuria significantly decreased in the corticosteroid group. Histological findings, such as mesangial cell proliferation, mesangial matrix accumulation, and cellular crescents, showed significant improvement in the corticosteroid group but not in the antiplatelet group. Expression of alphaSMA in glomeruli significantly decreased in the corticosteroid group but not in the antiplatelet group. EDA-FN did not change in either group. We conclude that early treatment with corticosteroids for adult diffuse proliferative IgA nephropathy is effective in reducing renal injury. PMID- 10676717 TI - ACE inhibition delays development of terminal renal failure in the presence of severe albuminuria. AB - The hypertensive fawn-hooded (FHH) rat develops progressive albuminuria (UalbV) and focal glomerulosclerosis (FGS). Early-onset angiotensin-converting enzyme inhibition (ACE-i) completely prevented the development of hypertension, UalbV, and FGS. ACE-i was still effective when the start of treatment was delayed, albeit less than early-onset treatment. In this study, we examined whether more advanced renal damage reduces the efficacy of ACE-i, and, if so, which factors dampen the efficacy. ACE-i was started in 36-week-old FHH rats, and follow-up consisted of regular assessment of systolic blood pressure (SBP) and UalbV. Untreated rats, matched for age, SBP, and UalbV, served as controls. In separate groups, untreated or treated with ACE-i from either week 7 or week 36, glomerular hemodynamics and FGS were determined at week 40. ACE-i normalized SBP and markedly reduced UalbV. The Initial UalbV response to ACE-i was inversely correlated with pretreatment UalbV, but despite control of SBP, UalbV rose again. Eventually, rats died of terminal renal failure. Life expectancy was significantly increased in treated rats. In both untreated and treated rats, there was a significant inverse correlation between baseline UalbV and survival time. However, the gain in survival time decreased when pretreatment UalbV was higher. Late-onset ACE-i reduced glomerular capillary pressure to the same extent as early-onset ACE-i. There was a significant linear correlation between FGS and UalbV. We conclude that in FHH rats with advanced renal damage, ACE-i slows down the progression to terminal renal failure. The outcome is an increased survival time that is inversely correlated with baseline UalbV. PMID- 10676718 TI - ACE gene polymorphism and survival in atherosclerotic renovascular disease. AB - Renovascular disease (RVD) is an important cause of end-stage renal disease and is associated with a high mortality rate, mostly because of coexisting cardiovascular and cerebrovascular disease. The deletion (DD) polymorphism of the angiotensin-converting enzyme (ACE) gene has been described in association with severe vascular disease affecting major organs. To investigate whether DD genotype is a risk factor for mortality in RVD, we performed a follow-up study of 61 patients with this disease. Patients (age, 68.0 +/- 6.5 years) affected by atherosclerotic vascular disease were enrolled after angiographic demonstration of a renal artery stenosis. The average follow-up was 48.1 +/- 14.9 months. Genotype was insertion/deletion (I/D) in 30 patients, DD in 27 patients, and II in 4 patients. At enrollment, a complete assessment of heart, blood vessels, and renal function was performed. During the follow-up period, 13 patients died (9 DD, 4 ID) and 7 patients evolved into end-stage renal failure. The cumulative survival rate at 5 years was 45.4% +/- 13.4%. Factors associated with mortality were analyzed with Cox proportional hazard regression. The multivariate analysis showed that DD genotype, severe carotid disease, and smoking were independent predictors of mortality. The multivariate analysis of predictors of renal failure showed that the only significant association was found with baseline serum creatinine level of 265 micromol/L or greater. We conclude that the DD genotype of the ACE gene is a marker for mortality in RVD. PMID- 10676719 TI - Digital glomerular reconstruction in a patient with a sporadic adult form of glomerulocystic kidney disease. AB - This study describes a sporadic adult form of glomerulocystic kidney disease in a 52-year-old man. To determine whether the aperture of the proximal tubule was stenosed or obstructed to clarify the pathogenesis of glomerular cyst development, 100 serial sections of the open biopsy specimen were made. Ten glomerular cysts were reconstructed using three-dimensional imaging analysis. Bowman's capsule (glomerular cyst) volume, the volume of glomerular tufts, and the area of the proximal tubular opening were estimated using imaging analysis. The volumes of Bowman's capsule and of glomerular tufts were 0.0098 +/- 0.0039 mm3 (mean +/- SD) (normal: 0.0041 to 0.0083 mm3) and 0.0026 +/- 0.0013 mm3, respectively. The area of the proximal tubular opening was 0.0017 +/- 0.0003 mm2 (normal: 0.0012 to 0.0028 mm2). There was neither obstruction nor stenosis of the opening of the renal tubule in this sporadic adult form of glomerulocystic kidney disease. After 4 years of hemodialysis, the glomerular cysts, as well as the kidneys, enlarged. This study shows that the main cause of glomerular cyst development is not glomerulotubular neck obstruction. PMID- 10676720 TI - Effect of probucol in a murine model of slowly progressive polycystic kidney disease. AB - Epithelial proliferation, extracellular matrix remodeling, and interstitial inflammation are central elements in the pathogenesis of slowly progressive polycystic kidney disorders. Probucol, an antioxidant that lowers plasma cholesterol, has been shown to decrease smooth muscle cell proliferation and macrophage accumulation in blood vessels and to prevent restenosis after coronary angioplasty. We determined in 30-day-old male BDF1-pcy hybrid mice (derived from mating DBA/2FG-pcy and C57BL/6FG-pcy) the effect of probucol administered in the diet (1%) for 200 days on kidney weight relative to body weight (KW/BW), cyst expansion, renal interstitial fibrosis, and serum urea nitrogen (SUN) concentration. Animals were fed a moderately high-protein diet (HPD, 36%) to accentuate the development of renal cysts and to promote interstitial fibrosis. Probucol decreased serum cholesterol from 68 to 16 mg/dL but had no effect on food intake or body weight. Probucol decreased relative kidney size from 4.16% +/ 0.55% to 2.64% +/- 0.12% KW/BW (P < 0.01), SUN from 30.5 +/- 1.8 to 25.9 +/- 1.0 mg/dL (P < 0.05), cystic index from 2.45 +/- 0.11 to 1.36 +/- 0.10 (P < 0.01), and fibrosis index from 2.40 +/- 0.11 to 1.82 +/- 0.08 (P < 0.01). We conclude that probucol ameliorates the progressive deterioration in renal function and structure in pcy mice ingesting a relatively high level of dietary protein. PMID- 10676721 TI - Effect of 1,25-dihydroxyvitamin D3 and calcium carbonate on bone loss associated with long-term renal transplantation. AB - To investigate the effect of calcitriol plus calcium carbonate on the bone loss associated with long-term renal transplantation, 30 patients with serum creatinine levels less than 2.0 mg/dL were randomly allocated to a control (n = 14) or treatment group (n = 16) and studied with bone biopsy and densitometry at baseline and after 1 year of follow-up. Calcitriol (0.25 microg/d) plus calcium carbonate (500 mg/d of elemental calcium) were administered to patients in the treatment group. Comparing the baseline and final data of each group at a time, no change in bone mineral density (BMD) z score was observed at the distal radius (control, -0.8 +/- 0.8 versus -0.6 +/- 0.9; treatment, -1.0 +/- 1.0 versus -1.0 +/- 1.1). However, a significant increase (P < 0.05) was found at the lumbar spine in both groups (control, 0.1 +/- 1.6 versus 0.4 +/- 1.6; treatment, -0.1 +/ 1.5 versus 0.3 +/- 1.5) and only in the treatment group at the femoral neck (control, -0.9 +/- 1.0 versus -0.8 +/- 1.0; treatment, -0.5 +/- 0.9 versus -0.3 +/- 1.1). When BMD was compared between groups, no significant differences were observed at the evaluated anatomic sites at baseline or after 1 year of follow up. After 1 year of follow-up, adjusting for age and sex (z score), the control group showed a trend to reduce the value of several histomorphometric parameters, including osteoblast surface (-2.2 +/- 6.1 versus -3.4 +/- 3.9), osteoid surface (-2.3 +/- 3.5 versus -3.1 +/- 3.9), and osteoclast surface (0.2 +/- 5.0 versus 1.3 +/- 3.3). Consequently, there was a significant reduction (P < 0.05) in mineralizing surface (-9.8 +/- 11.0 versus -15.8 +/- 12.3) and appositional rate (-5.8 +/- 2.7 versus -7.6 +/- 2.2). In the treatment group, a significant reduction (P < 0.05) in osteoclast surface was observed at the end of the study (3.9 +/- 6.8 versus -1.2 +/- 4.1), and although a trend to reduce osteoblast surface (-2.5 +/- 2.6 versus -3.2 +/- 5.7) and osteoid surface (-2.1 +/- 2.5 versus -3.2 +/- 2.8) was also found, patients maintained approximately the same level of wall thickness (-5.2 +/- 5.3 versus -5.3 +/- 3.3) and bone volume (-2.7 +/- 1.8 versus -2.5 +/- 1.7). However, there was no improvement in mineralizing surface (-4.2 +/- 2.9 versus -10.4 +/- 3.6) or appositional rate (-5.8 +/- 3.1 versus -8.1 +/- 2.6). No significant differences in bone histomorphometric variables were observed between groups after 1 year of follow-up. In conclusion, 1,25-dihydroxyvitamin D3 and calcium carbonate did not significantly improve bone loss in long-term renal transplant recipients. However, significant osteoclast suppression and a trend to maintain trabecular bone volume and wall thickness as well as improve the axial BMD were observed in the treatment group. PMID- 10676722 TI - Life expectancy benefits of cancer screening in the end-stage renal disease population. AB - Health maintenance includes secondary prevention through cancer screening. There are no established guidelines for cancer screening patients with end-stage renal disease (ESRD). Using an established method of estimating life expectancy, published literature on cancer screening, and information from databases on mortality and malignancy (US Renal Data System 1997 Annual Data Report and the SEER Cancer and Statistical Review, 1973-1994), a "real-time life expectancy calculator" was developed to guide the primary help provider in making informed decisions on the benefits of cancer screening in individual patients. Potential days of life saved by each screening method can be calculated using the difference in life expectancy per the DEALE (declining exponential approximation of life expectancy) method with and without cancer screening. Using two sets of assumptions (one to enhance any bias toward support for screening and one to limit this bias), a range of potential days of life saved with screening for breast and colon cancer can be calculated in individual patients with ESRD. In breast cancer, for example, a 50-year-old black woman with ESRD and multiple risk factors would have 41 to 291 potential days of life saved with screening. A 60 year-old white woman with ESRD and diabetes mellitus (DM) would have only 1 to 16 days of life saved. This life expectancy calculator can guide the primary health care provider in making clinical decisions concerning screening in the ESRD population. In addition to assisting in patient education, the calculator can be updated as new information becomes available regarding relative risk, treatment, and mortality. PMID- 10676723 TI - Impact of disease severity and hematocrit level on reuse-associated mortality. AB - Prior studies on reuse-associated mortality have presented conflicting results and included few adjustments for disease severity or hematocrit levels. To evaluate the impact of patient and provider characteristics on reuse-associated mortality, we developed a period-prevalent model with a 6-month entry period. Five cohorts of Medicare hemodialysis patients surviving from July 1 through December 31 of the entry year (1991, 60,985 patients; 1992, 63,081 patients; 1993, 76,018 patients; 1994, 82,899 patients; 1995, 91,761 patients) were followed up for the next year. Using a basic Cox regression survival model (M-1) including age, sex, race, renal diagnosis, prior end-stage renal disease time, unit age, unit size, water treatment, dialysate, and germicide, results were compared with those using a more inclusive model (M-4) adding dialyzer type (conventional or high efficiency/high flux), unit designation (hospital based or freestanding), unit profit status, comorbidity, disease severity, and hematocrit. The previous association of for-profit units with increased mortality was not present after 1994. Whereas the M-1 analysis showed better survival in reuse units after 1991, the more complete M-4 analysis showed no difference in the risk for mortality between reuse and no-reuse units. We conclude that mortality rates in the United States from 1991 to 1995, when adjusted comprehensively for patient and unit characteristics, were not different in units that practiced reuse and those that did not. PMID- 10676724 TI - Cardiovascular effect of normalizing the hematocrit level during erythropoietin therapy in predialysis patients with chronic renal failure. AB - The optimal target hematocrit (Ht) level in recombinant human erythropoietin (rHuEPO) therapy remains controversial and has hardly been investigated in predialysis patients. We prospectively studied the regression of left ventricular hypertrophy (LVH) on echocardiography in nine predialysis patients with chronic renal failure after a partial correction (target Ht, 30%) and normalization (target Ht, 40%) of the Ht with rHuEPO treatment. Twenty-four-hour ambulatory blood pressure monitoring was also performed. The administration of rHuEPO significantly increased Ht to the target values. The rate of renal failure progression did not change during rHuEPO treatment for 12 months (Cr, from 6.2 +/ 2.0 to 5.5 +/- 2.1 mg/dL). The left ventricular mass index (LVMI) tended to decrease after a partial correction of anemia (Ht, 32.1% +/- 1.8%) at 4 months, whereas it tended to significantly decrease after normalization of Ht (Ht, 39.1% +/- 2.4%) at 12 months (baseline, 140.6 +/- 12.1 g/m2; partial correction, 126.9 +/- 10.0 g/m2; normalization, 111.2 +/- 8.3 g/m2). All patients had received antihypertensive medication before rHuEPO administration, and additional drugs were also required in four cases during the study. As a result, a good overall blood pressure control was obtained without any adverse effects on the circadian blood pressure rhythm. In conclusion, from the perspective of LVH regression, the normalization of Ht was found to be more effective than that associated with a partial correction of anemia during rHuEPO therapy. PMID- 10676725 TI - Interdialytic weight gain, compliance with dialysis regimen, and age are independent predictors of blood pressure in hemodialysis patients. AB - Hypertension is a common problem in patients undergoing chronic hemodialysis. The purpose of this study is to identify the clinical and demographic factors independently associated with blood pressure in this population. Data collected for the Dialysis Morbidity and Mortality Study Wave 1 by the US Renal Data System were analyzed. The mean predialysis blood pressure for this cohort of 5,369 patients was 149/79 mm Hg. Sixty-three percent of the patients were hypertensive; 27%, 25%, and 11% had stages 1, 2, and 3 hypertension, respectively. Young age, black race, male sex, diabetes as cause of end-stage renal disease, erythropoietin therapy, and smoking were associated with higher blood pressure in the univariate analysis. Patients skipping or shortening one or more dialysis treatments had higher blood pressure. The presence of congestive heart failure and coronary heart disease was associated with lower blood pressure. On multivariate analysis, high interdialytic weight gain, noncompliance with dialysis regimen, and younger age were independent predictors of higher blood pressure. In summary, hypertension is common and poorly controlled in patients undergoing chronic hemodialysis. Greater interdialytic weight gain and noncompliance with dialysis regimen are independently associated with higher blood pressure, and advancing age is associated with lower blood pressure levels in this population. Therapeutic regimens emphasizing reduction of interdialytic weight gain and improved compliance with the dialysis regimen need to be evaluated for improving the management of hypertension. The effect of age and other comorbid conditions, particularly cardiovascular disease, must be considered while studying the relationship between blood pressure and mortality in patients undergoing chronic hemodialysis. PMID- 10676726 TI - Improved urea reduction ratio and Kt/V in large hemodialysis patients using two dialyzers in parallel. AB - Delivered dose of hemodialysis (HD) in large patients with end-stage renal disease is often less than adequate. Fourteen chronic HD patients with weights greater than 80 kg participated in a prospective, cross-over study comparing urea reduction ratio (URR +/- SEM) and the fractional clearance index for urea (eKt/V(urea) +/- SEM) on a single polysulfone dialyzer for a control (HDC) period of 4 weeks versus clearances obtained with two dialyzers in parallel during an intervention (HDP) period of 4 weeks. Clearance of the surrogate middle molecule iohexol (C(Io)) was also measured. Health status was assessed with the SF-36. Blood and dialysate flow rates and duration of HD sessions were constant. URR increased from 0.67 +/- 0.006 during HDC to 0.72 +/- 0.006 with HDP (P < 0.0001). eKt/V(urea) increased from 1.16 +/- 0.021 to 1.34 +/- 0.021 (P < 0.0001). Increased URR and eKt/V(urea) occurred in all 14 during HDP (P < 0.05). C(Io) during HDP averaged 182 +/- 7.7 mL/min compared with 131 +/- 5.4 mL/min in HDC sessions (P < 0.00001). Health status improved in six of eight categories. Expense increased approximately $14.27 per dialysis with HDP. In 11 of 14 patients continued on two dialyzers in parallel for 1 year, monthly eKt/V averaged 1.46 +/- 0.066, and health status further improved in five of eight categories. In large patients, two dialyzers in parallel increased urea and iohexol clearance. Increased urea clearance was maintained for 1 year, and health status improved. PMID- 10676727 TI - Use of standardized ratios to examine variability in hemodialysis vascular access across facilities. Medical Review Board of The Renal Network, Inc. AB - The type of hemodialysis vascular access (catheter, fistula, graft) is an important determinant of patient morbidity and dialysis efficiency. The relative importance of patient versus provider factors in determining type of vascular access is unclear. We sought to develop a quality improvement tool that adjusts for differences in patient characteristics, thereby allowing examination of provider-related variability in types of vascular access used across facilities. We examined 15,339 patients from 216 chronic hemodialysis units in Indiana, Kentucky, Ohio, and Illinois and found that 20% of patients had catheters, 24% had fistulas, and 56% had grafts. Young, male, and white patients were more likely to have fistulas, whereas old, female, and black patients were more likely to have grafts. Diabetics were more likely to have catheters and less likely to have fistulas. New patients were more likely to have catheters and less likely to have grafts. A facility specific standardized catheter ratio (SCR), standardized fistula ratio (SFR), and standardized graft ratio (SGR) were calculated based on the actual number of patients with each type of vascular access divided by the expected number adjusted for patient characteristics. Facility SCRs ranged from 0.00 to 2.87. Of the 216 facilities, 38 (18%) had an SCR significantly less than 1.00, and 32 (15%) had an SCR significantly greater than 1.00. Similar variability was observed in SFRs and SGRs. In conclusion, the type of vascular access varies greatly across facilities. Use of standardized access ratios adjusted for patient characteristics may help providers examine processes of care that contribute to variability in access use. Analogous to the standardized mortality ratio, the SCR, SFR, and SGR should be effective quality improvement tools. PMID- 10676728 TI - Septicemia in diabetic hemodialysis patients: comparison of incidence, risk factors, and mortality with nondiabetic hemodialysis patients. AB - Diabetes mellitus is the most common cause of treated end-stage renal disease (ESRD), and diabetic hemodialysis patients have a high mortality rate. To identify differences in risk of septicemia among diabetic and nondiabetic hemodialysis patients, we examined the incidence, risk factors, and mortality for septicemia in a large sample of the US hemodialysis population. We performed a longitudinal cohort study of the incidence and risk factors for hospitalized cases of septicemia in diabetic and nondiabetic hemodialysis patients using baseline data from the US Renal Data System case-mix severity study with 7-year follow-up from hospitalization and death records. Independent risk factors for septicemia were assessed using Poisson regression. Independent effect of septicemia on mortality was assessed using Cox proportional hazards analysis. Over 7 years, 11.1% of nondiabetic patients and 12.5% of diabetic patients experienced at least one episode of septicemia. Older age and low serum albumin were independent risk factors for septicemia in all patients. In diabetics, white race, peripheral vascular disease, and hemodialyzer reuse, particularly in type 1, were independent risk factors. In nondiabetics, coronary artery disease, cerebrovascular disease, and temporary and permanent catheters were associated with an increased risk. In both groups, patients who experienced an episode of septicemia had twice the risk of death from any cause and an eightfold risk of death from septicemia. Septicemia occurs equally frequently and carries a marked increased risk of death in both nondiabetic and diabetic hemodialysis patients. Improving nutritional status and minimizing the use of catheters might help ameliorate the risk of septicemia. In diabetics, aggressive treatment of peripheral vascular disease might help reduce the risk of septicemia. Further research to elucidate potential mechanisms for variations in risk for septicemia according to race and hemodialyzer reuse practices are warranted in diabetic patients. PMID- 10676729 TI - Quality-of-life evaluation using Short Form 36: comparison in hemodialysis and peritoneal dialysis patients. AB - Short Form 36 (SF-36) is a well-documented health-related quality-of-life (HRQOL) instrument consisting of 36 questions compressed into eight scales and two primary dimensions: the physical and mental component scores. This tool was used to evaluate QOL among peritoneal dialysis (PD) and hemodialysis (HD) patients. The results of 16,755 HD and 1,260 PD patients (728 continuous ambulatory PD [CAPD] and 532 continuous cycling PD [CCPD]) completing an SF-36 during 1996 were analyzed. Three analyses of variance were performed, consisting of (1) no adjustment, (2) case mix (age, sex, race, and diabetes), and (3) case mix plus laboratory parameters. PD patients were younger (P < 0.001), a larger fraction were white (P < 0.001), fewer had diabetes (P < 0.001), and had lower serum albumin concentrations (P < 0.001) and higher creatinine, hemoglobin, and white blood cell count values (P < 0.001) than HD patients. Diabetes was present in a larger fraction of CCPD than CAPD patients (P < 0.001). HD and PD patients scored similarly for scales reflecting physical processes. PD patients scored higher for mental processes, but only after statistical adjustment for the laboratory measures. Scores on scales reflecting physical processes were worse, and those reflecting mental processes were better among CCPD than CAPD patients. HD and CAPD scores were similar. CCPD patients perceived themselves as more physically impaired but better adjusted than HD or CAPD patients. These descriptive data show that perception of QOL among PD and HD patients is similar before adjustment, but PD patients score higher for mental processes with adjustment. CCPD patients score worse for physical function and better for mental function than either CAPD or HD patients. We cannot, however, exclude the influence of therapy selection. PMID- 10676730 TI - Role of Fogarty catheter manipulation in management of migrated, nonfunctional peritoneal dialysis catheters. AB - Peritoneal dialysis (PD) catheter migration to the upper abdomen is not an uncommon cause of catheter failure. We prospectively examined the role of the Fogarty catheter manipulation technique to reposition the PD catheter in the pelvis and regain patency. All patients with PD catheter malfunction caused by migration, confirmed by abdominal radiograph, underwent the same protocol. The patient was placed flat on the back, and the Fogarty was advanced into the PD catheter to a premarked point at which the end of the Fogarty was near the end of the PD catheter. The Fogarty balloon was inflated with 0.5 mL of sterile saline, and manipulation was performed by tugging movements until proper placement of the PD catheter into the pelvis was suspected. Infusion and drainage of dialysate was performed to determine patency. The return of the PD catheter into the pelvis was then confirmed by repeated radiograph. Success rates of Fogarty catheter manipulation, early and late recurrence (remigration < or =90 days or >90 days), and complications were prospectively examined in 232 patients over a 6-year period. Catheter migration occurred in 34 of 232 patients (15% incidence). All patients had curled-end, double-cuffed, non-swan-neck PD catheters. Successful repositioning occurred in 24 of 34 patients (71%). None of the 24 repositioned catheters had early recurrence, and 1 of 24 catheters (4%) had late recurrence. None of the patients had procedure-related peritonitis, bowel perforation, or exit-site trauma. These results show that PD catheter migration is relatively common (15%). The Fogarty manipulation technique is a simple, cost-effective way to prolong PD catheter life and preserve its long-term patency. This eliminates the need for surgical intervention in approximately 70% of patients with PD catheter migration. PMID- 10676731 TI - Mouse to elephant: biological scaling and Kt/V. AB - The construct Kt/V is used by the nephrology community in prescribing dialysis dose. The concerns that have been raised as to what value of V to use in the calculation of Kt/V touch on the more central question of whether filtration rate should be normalized by a parameter other than V. Within the animal kingdom, a number of physiological variables scale to body size according to an equation of the form Y = YoMb, where Yo is a constant, M is body mass, and b is a scaling exponent. Glomerular filtration rate (GFR) in mammals weighing from 30 g to 503 kg scales to body weight with an exponent of 0.77. Hence, GFR per unit body weight (or Kt/V) decreases significantly with increasing body size. Metabolic rate also scales to body size in a wide range of mammals according to the same general equation and with a scaling exponent of 0.75. Because GFR and metabolic rate scale to body mass with virtually the same exponent, a ratio of the two yields a constant independent of body size. We propose that the ratio (filtration rate/metabolic rate) replace Kt/V. Such a ratio would underscore the linkage between filtration rate (and dialysis therapy) and the metabolic demands of the body. PMID- 10676732 TI - Noni juice (Morinda citrifolia): hidden potential for hyperkalemia? AB - We report the case of a man with chronic renal insufficiency who self-medicated with an alternative medicine product known as noni juice (Morinda citrifolia). The patient presented to the clinic with hyperkalemia despite claiming adherence to a low-potassium diet. The potassium concentration in noni juice samples was determined and found to be 56.3 mEq/L, similar to that in orange juice and tomato juice. Herbal remedies and alternative medicine products may be surreptitious sources of potassium in patients with renal disease. PMID- 10676733 TI - Rapidly progressive fibrosing interstitial nephritis associated with Chinese herbal drugs. AB - Rapidly progressive fibrosing interstitial nephritis after a slimming regimen containing aristolochic acid has been identified as Chinese herbs nephropathy (CHNP). From 1995 to 1998, we observed 12 Chinese people from different areas of Taiwan who underwent renal biopsy for unexplained renal failure. Medical history gave no clue to the causes of impaired renal function except for the ingestion of traditional Chinese herbs. Although these patients ingested herbal drugs from various sources for different purposes, their renal biopsy samples showed amazingly similar histological findings, with extensive hypocellular interstitial fibrosis and atrophy and loss of tubules in all cases. Glomeruli were apparently intact. They also had similar clinical features, such as normal or mildly elevated blood pressure, early and severe anemia, low-grade proteinuria, glycosuria, and insignificant urinary sediments. Renal function deteriorated rapidly in most patients despite discontinuation of the herbal medicines. Seven patients underwent dialysis, and the remainder experienced slowly progressive renal failure. Bladder carcinoma was found in one patient. Morphologically and clinically, the nephropathy in our patients was similar to CHNP, reported in Belgium. Because of the complexity and unknown types of herbs used in different clinical situations, unidentified phytotoxins other than aristolochic acid might be responsible for this unique disease entity. We conclude that the relation of this nephropathy to the consumption of Chinese herbs is striking. Using uncontrolled herbal remedies carries a high risk for developing interstitial renal fibrosis and urothelial malignancy. PMID- 10676734 TI - Cardiopulmonary manifestations of Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is usually a mild condition involving the skin, gut, joints, and kidneys and has a good prognosis. We present a 63-year-old Hispanic man who had an unusually severe form of HSP with a fatal outcome attributable to vasculitis causing myocardial necrosis. There is only one citation in the literature of HSP-related myocardial vasculitis, which involved the right ventricle and was successfully treated with steroids. Our patient had severe HSP-related myocardial necrosis, tracheobronchitis, and nephritis. The bronchial lesions resolved, presumably because of steroid therapy. This probably is the first case of fatal myocardial necrosis related to HSP. We conclude that HSP can, in some cases, have an aggressive course. It becomes imperative to recognize the involvement of the other organ systems, such as the heart, so that appropriate therapy may be initiated. Immunosuppression may have a beneficial effect on extrarenal lesions. Controlled clinical trials are needed to establish the efficacy of such treatment. PMID- 10676735 TI - Charcoal hemoperfusion in the treatment of phenytoin overdose. AB - In the case of phenytoin, a drug that is generally highly protein bound, there is a lack of consensus on the use of charcoal hemoperfusion in cases of overdose. We performed charcoal hemoperfusion on a phenytoin-overdosed patient to assess the effectiveness of this treatment. The plasma concentrations of total and free phenytoin fell rapidly, from 40.0 microg/mL and 3.6 microg/mL to 16.2 microg/mL and 1.5 microg/mL, respectively, after 3 hours of hemoperfusion. The total phenytoin elimination half-life was 3.9 hours. The fraction of protein-bound phenytoin was constant (90.8% +/- 0.5%) before, during, and after the procedure. The relations between the in vitro protein binding and adsorption of phenytoin to activated charcoal were also examined. Interestingly, bound phenytoin was found to dissociate from plasma proteins in the presence of activated charcoal and subsequently became adsorbed to the activated charcoal. Considering that phenytoin is bound to albumin with a large number of binding sites (n = 6) and a small binding constant (K = 6 x 10(3/)mol/L), the extent of adsorption to activated charcoal may depend on the magnitude of the binding constant of the drug to plasma proteins. The current results suggest that charcoal hemoperfusion is effective for the removal of drugs that bind to plasma proteins with a low binding constant. PMID- 10676736 TI - The failure of breast cancer screening. PMID- 10676737 TI - Nephropathy and herbal medicine. PMID- 10676738 TI - Nephrotoxicity of immunosuppressive drugs: long-term consequences and challenges for the future. AB - The calcineurin inhibitors cyclosporin A (CsA) and tacrolimus (FK506) are associated with dose- and efficacy-limiting adverse events, including nephrotoxicity, which may diminish their overall benefits for long-term graft survival. Nephrotoxicity is difficult to distinguish from chronic allograft rejection and is a particular problem in the setting of renal transplantation. Minimizing immunosuppressant-induced nephrotoxicity could improve long-term renal allograft survival. However, to obtain significant long-term improvement in renal allograft outcomes, it may be necessary to adopt new immunosuppressive regimens that rely less on calcineurin inhibitors. Recipients of other transplanted organs, as well as patients with autoimmune diseases who require immunosuppressant therapy, could also benefit from this change in immunosuppressive drug strategy because their healthy, native kidneys are particularly susceptible to the nephrotoxic effects of CsA and FK506. CsA- and FK506-sparing regimens, which use reduced doses of CsA and FK506 in combination with other nonnephrotoxic immunosuppressants, may be the best current option for reducing nephrotoxicity. The chemical immunosuppressant mycophenolate mofetil (MMF) has been used as part of CsA- and FK506-sparing regimens that provide improved renal function while maintaining adequate immunosuppression. Such regimens should reduce patient morbidity and mortality. Also, because immunosuppressant-induced nephrotoxicity has been associated with significant financial costs, CsA- and FK506-sparing regimens should result in substantial savings in health care costs. PMID- 10676739 TI - Incidence and spectrum of dialysis-associated cancer in three continents. PMID- 10676740 TI - Staphylococcus aureus pneumonia, hyponatremia, hypertension, proteinuria, and hematuria in a 14-year-old boy. PMID- 10676741 TI - Pathogenesis of anaphylactoid reactions to intravenous iron. PMID- 10676742 TI - Prolonged slow dialysis and better survival. PMID- 10676743 TI - Mandatory cardiac surgery in a patient with a recent history of cholesterol crystal embolism. PMID- 10676744 TI - Identical glomerulopathy in two different mouse models of uteroglobin deficiency. PMID- 10676745 TI - Does imbalance between basal ganglia and cerebellar outputs cause movement disorders? PMID- 10676746 TI - Normal pressure hydrocephalus: developments in determining surgical prognosis. AB - Research into normal pressure hydrocephalus has often focused on the clinical dilemma of selecting patients who will benefit from cerebrospinal fluid diversion. Recent developments in imaging and lumbar infusion tests are throwing light on the underlying pathophysiology, providing researchers with new avenues for the development of reliable investigative tools. PMID- 10676747 TI - Neoplasms. PMID- 10676748 TI - Medulloblastoma. AB - The utilization of multi-modal therapy in the treatment of medulloblastoma has improved survival rates and overall outcome. Recent large clinical trials have supported the use of radiation and chemotherapy as adjuvant treatment. Treatment advances have been made despite a poor understanding of the biological underpinnings of medulloblastoma. Current laboratory investigations are shedding light on the oncogenesis of medulloblastoma and may lead to improved treatments. PMID- 10676749 TI - Primary central nervous system lymphoma. AB - Primary central nervous system (CNS) lymphoma is an unusual but increasingly frequent brain tumor being identified by neurologists. Considerable improvements in survival have been accomplished by the addition of chemotherapy to cranial radiotherapy. In addition, many patients achieve substantial disease free survival with chemotherapy alone, and survival is superior to radiotherapy alone. Currently, every patient should be considered for chemotherapy as the first line of treatment. Subsequent cranial radiotherapy may or may not be necessary depending upon the patient's clinical condition, age and response to initial chemotherapy. Intensification of multi-agent chemotherapy is under study and it is anticipated that the incidence of neurotoxicity will reduce with elimination of combined modality treatment. PMID- 10676750 TI - Recent advances in the diagnosis and treatment of central nervous system germ cell tumours. AB - Primary germ-cell tumors of the central nervous system are rare neoplasms that are seen primarily in the pediatric age group. Because of their frequent location in the pituitary and suprasellar regions, they present with typical neuro ophthalmologic and neuroendocrine symptoms. Sophisticated imaging and surgical biopsy allow precise anatomic definition, but only allow an approximate guess of the tumor histopathology. Tumor markers in the serum and cerebrospinal fluid are extremely helpful in the diagnosis and monitoring of response to treatment when they are detectable. Because of the deleterious effects of irradiation on neurocognitive and neuroendocrine functioning, we have looked at strategies that either reduce or eliminate radiation exposure. Large, randomized, prospective, cooperative trials in the future will be the only way to identify subgroups of patients that may benefit from particular treatment strategies. PMID- 10676751 TI - Rationales for improving motor function. AB - New findings in basic neuroscience, and the growing knowledge regarding neuroplasticity and motor learning have exerted influence and have provided stimuli for motor rehabilitation research. Repeated motor practice has been identified as crucial for motor recovery. Further novel and scientifically based therapeutic approaches have been developed: constraint-induced movement therapy, electromyogram-initiated neuromuscular stimulation, motor imagery and music therapy are all discussed in the present review. PMID- 10676752 TI - Setting goals for cognitive rehabilitation. AB - Evidence for experience-dependent plasticity of the brain, including cell regeneration, means that rehabilitation can aim at reinstituting impaired cognitive function, as well as at training compensatory strategies for the lost function. New theoretical frameworks make predictions regarding the circumstances under which these two approaches should each be attempted. There has been progress over the past 6 years in designing effective rehabilitation strategies, with more of these having a strong theoretical basis in cognitive neuroscience. Basic cognitive science has generated counter-intuitive, but effective cognitive rehabilitation methods, showing that the goal of rehabilitation need not always be the most obvious one dictated by real life performance. Limb Activation Training for unilateral neglect is an example of a theoretically derived cognitive rehabilitation procedure that has now been clinically evaluated in clinical trials. PMID- 10676753 TI - Mapping plastic brain changes after acute lesions. AB - The combination of different mapping techniques has yielded new insights in reorganization processes after acute lesions in humans. Recent research focused not only on lesion-induced plasticity, but also on therapy-induced reorganization of the brain. Data from animal experiments has expanded our knowledge of mechanisms that underlie plastic changes. PMID- 10676754 TI - Experimental models of brain trauma. AB - A short review of the most widely used and popular experimental models of traumatic brain injury is presented. This review focuses on current animal models of traumatic brain injury that apply mechanical energy to the skull or, after trephination of the skull, to the intact dura. Recent experimental studies evaluating the pathobiology of traumatic brain injury using these models are also discussed. This article attempts to provide a broad overview of current knowledge and controversies in experimental animal research on brain trauma. PMID- 10676755 TI - Metabolic disorders and neurotoxicology. PMID- 10676756 TI - The pathogenesis of neurodegenerative disease: neurotoxic mechanisms of action and genetics. AB - The role of environmental and occupational exposures to neurotoxicants in the pathogenesis of neurodegenerative disease has not been fully elucidated. Recent published research on whether genetic polymorphisms contribute to individual susceptibility to develop neurodegenerative diseases such as Parkinson's disease have been equivocal at best. This review relates putative mechanisms of neurotoxicant-induced cell damage to polymorphisms in the genes that encode for the enzymes involved in the metabolism of neurotoxicants. The effects that genetically induced alterations in enzyme functioning have on neurotoxicant metabolism and how this relates to the risk of neurotoxic effects among exposed individuals are reviewed. A pragmatic approach to future research in the area of neurodegenerative disease is developed on the basis of the interrelationship between known routes of neurotoxicant metabolism and human genetics. PMID- 10676757 TI - Neurotoxicants and the cytoskeleton. AB - Exposure to occupational and environmental toxicants can result in distal axonopathies through reaction with various components of the axonal cytoskeleton. The solvents n-hexane and methyl n-butyl ketone are metabolized to the beta diketone, 2,5-hexanedione, which covalently cross-links neurofilaments, resulting in large paranodal axonal swellings filled with neurofilaments. Carbon disulfide exposure leads to an identical axonopathy, achieving neurofilament cross-linking through a parallel series of reactions. Acrylamide and ethylene oxide, on the other hand, adduct proteins but do not lead to cross-linking. These toxicants appear to affect the function of microtubule-associated proteins, such as kinesin, and result in the impaired transport of synaptic vesicles. PMID- 10676758 TI - Tracking cholinergic pathways from psychological and chemical stressors to variable neurodeterioration paradigms. AB - Cholinergic hyperexcitation can be induced by both acute psychological stress and exposure to acetylcholinesterase inhibitors. Both factors are known risk factors for delayed neurodeterioration processes such as Alzheimer's disease and Parkinson's disease. Recent publications on the involvement of cholinergic pathways in these and other neurodeterioration syndromes are reviewed. PMID- 10676759 TI - Neurotoxic effects of endocrine disruptors. AB - Endocrine disrupting chemicals are a newly defined category of environmental contaminants that may affect animal and human populations by interfering with normal hormone action. There is substantial concern that these agents could have a range of subtle and long-lasting effects. Because of the sensitivity of the developing central nervous system to low levels of endogenous gonadal hormones during development, the central nervous system may be a target for the action of endocrine disrupting chemicals. PMID- 10676760 TI - Magnetic resonance spectroscopy in toxic encephalopathy and neurodegeneration. AB - Magnetic resonance spectroscopy allows neurochemistry to be probed noninvasively in vivo. Recent advances in our understanding of the biochemical significance of the various neurochemicals that are observable allow a variety of pathologic states of relevance to encephalopathies and neurodegenerative disorders to be observed. Measurements of brain glutamate and glutamine allow observation of neuronal/glial substrate cycling and ammonia detoxification. Myo-inositol allows changes in cerebral osmolarity and gliosis to be observed. N-acetylaspartate is a marker of neuronal health and number. Lactate allows nonoxidative glycolysis to be observed. These molecules are now being used to ask etiologic questions that are of relevance to encephalopathies and neurodegeneration, as well to probe longitudinally both natural history and therapeutic interventions in these conditions. Combined with recent advances in anatomic magnetic resonance imaging as well as perfusion magnetic resonance imaging, magnetic resonance spectroscopy has the potential to aid greatly in our understanding of neuronal dysfunction in a wide variety of neurologic pathologies, even in single patients. PMID- 10676761 TI - Multiple antioxidants in the prevention and treatment of neurodegenerative disease: analysis of biologic rationale. AB - Parkinson's disease and Alzheimer's disease are major progressive neurologic disorders, the risk of which increases with advancing age (65 years and over). In familial cases, however, early onset of disease (35-65 years) is observed. In spite of extensive basic and chemical research on Parkinson's disease and Alzheimer's disease, no preventive or long-term effective treatment strategies are available. The analysis of existing literature suggests that oxidative stress is a major intermediary risk factor for the action of diverse groups of neurotoxins that are involved in these neurodegenerative diseases. In this review, it is proposed that the epigenetic components (mitochondria, other organelles, membranes, protein modification) rather than nuclear genes of neurons are the primary targets for the action of neurotoxins, including free radicals. In addition, a scientific rationale for using multiple antioxidants in clinical trials for the prevention of Parkinson's disease and Alzheimer's disease among high-risk populations, and as an adjunct to standard therapy in the treatment of these diseases is presented. PMID- 10676762 TI - Bibliography. Current world literature. Degenerative diseases. PMID- 10676763 TI - Bibliography. Current world literature. Neoplasms. PMID- 10676764 TI - Bibliography. Current world literature. Trauma and rehabilitation. PMID- 10676765 TI - Bibliography. Current world literature. Metabolic disorders and neurotoxicology. PMID- 10676766 TI - Hematology and oncology PMID- 10676767 TI - Idiopathic thrombocytopenic purpura: beyond consensus. AB - Idiopathic thrombocytopenic purpura (ITP) is the most common acquired bleeding disorder encountered by pediatricians. Most children with ITP have minimal bleeding and complete platelet count recovery within weeks to months. Therapy for ITP has ranged from close observation without medical intervention to aggressive management with corticosteroids, intravenous immunoglobulin G, or anti-D immune globulin. The topic of ITP has incited great debate among practitioners, and this debate prompted the development of ITP practice guidelines by the British Paediatric Haematology Group in 1992 and by the American Society of Hematology in 1996. A better understanding of the clinical course of, risk for significant bleeding in, and optimal evaluation and therapy of childhood ITP will require carefully designed, multicenter, clinical trials. PMID- 10676768 TI - Oncology practice patterns in the use of hematopoietic growth factors. AB - Recombinant hematopoietic growth factors were introduced into clinical practice a decade ago: erythropoietin in 1989, granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in 1991, and interleukin-11 in 1997. The role of these agents in supportive therapy for children with cancer is still under considerable evaluation. This pediatric-based review summarizes current clinical applications, practice guidelines, and practice patterns for hematopoietic growth factors in the supportive care of children with cancer. It also discusses ongoing controversies and unanswered questions. PMID- 10676769 TI - Return of granulocyte transfusions. AB - Recently, several groups have begun to administer granulocyte colony-stimulating factor (G-CSF), a hematopoietic growth factor, with or without dexamethasone to mobilize peripheral blood neutrophils. Granulocyte colony-stimulating factor (600 microg subcutaneously) and dexamethasone (8 mg orally) given 12 hours before standard leukapheresis routinely results in the collection of approximately 80 x 10(9) granulocytes. This number of cells is sufficient to increase the neutrophil count of a severely neutropenic patient to normal and restore the recipients' ability to develop a neutrophil response in tissues. Several trials are ongoing to establish the clinical benefit of this new approach to supportive care for neutropenic patients. PMID- 10676770 TI - Thrombotic thrombocytopenic purpura and the hemolytic-uremic syndrome. AB - Large and unusually large von Willebrand factor (vWf) multimers may be responsible for systemic platelet aggregation in thrombotic thrombocytopenic purpura (TTP). This possibility is supported by studies that show deficient vWf cleaving metalloproteinase and increased platelet-vWf binding during TTP episodes. In acute idiopathic TTP, decreased vWf metalloproteinase is the result of autoantibodies against the enzyme. In familial and acquired hemolytic-uremic syndrome, vWf-cleaving metalloproteinase activity is normal. A deficiency or defect in factor H, which normally dampens the activation of C3 via the alternative complement pathway, has been seen in some patients with familial hemolytic-uremic syndrome. Ticlopidine therapy is an important risk factor for TTP. PMID- 10676771 TI - Congenital and inherited polycythemia. AB - Absolute polycythemia is a condition with increased red blood cell mass. There are a number of primary and secondary polycythemic disorders leading to absolute polycythemia. Primary polycythemias are caused by a defect intrinsic to the erythroid progenitor cells. The best characterized primary polycythemia is the autosomal dominant primary familial and congenital polycythemia (PFCP). Familial or childhood occurrence of the myeloproliferative disorder polycythemia vera are also discussed, emphasizing the importance of distinction between polycythemia vera and PFCP. Congenital or familial secondary polycythemic conditions are characterized by increased red cell mass, which is caused by circulating serum factors, typically erythropoietin. PMID- 10676772 TI - More than skin deep. PMID- 10676773 TI - Perinatal brachial plexus palsy. AB - Perinatal brachial plexus palsy (PBPP) has been traditionally classified into three types: upper plexus palsy (Erb's) affecting the C5, C6, and +/- C7 nerve roots, lower plexus palsy (Klumpke's) affecting the C8 and T1 nerve roots, and total plexus palsy. Although most cases will resolve spontaneously, the natural history of the remaining cases is influenced by contractures of uninvolved muscle groups and subluxation or dislocation of the shoulder and elbow. Microsurgical nerve repair has demonstrated to provide improved outcomes compared to conservative treatment, while advancements in secondary reconstruction have offered significant improvements in the performance of activities of daily living for older children with unresolved plexus palsy. PMID- 10676774 TI - Transient synovitis as a cause of painful limps in children. AB - Transient synovitis of the hip is one of the most common causes of hip pain and limp in young children. Its cause is still largely unknown, but its natural history is that of a self-limiting disorder with no residual sequelae, although recurrences are possible. Because this benign condition is similar to more significant disorders, such as septic arthritis, the diagnosis should remain one of exclusion. Once transient synovitis is diagnosed, treatment consists of rest, anti-inflammatory agents, and a tincture of time. PMID- 10676775 TI - Ankle injuries in the pediatric population. AB - Ankle injuries occur with considerable frequency in the pediatric population, although diagnosis is rarely straightforward. In this paper, we highlight the more common pitfalls in diagnosis and management, calling upon classic and contemporary literature and our own combined 30-year experience in managing pediatric lower extremity problems. PMID- 10676776 TI - Congenital scoliosis. AB - Congenital scoliosis is a deformity of the developing spine that results from defects in vertebral development. The developmental etiologies may be classified as either a failure of formation, a failure of segmentation, or a mixture of these two modes of maldevelopment. Early detection and close surveillance of congenital scoliosis is critical, as a rapidly progressive curve may lead to significant deformity, pulmonary restriction, and neurologic problems if not treated. Early surgical treatment is often necessary for rapidly progressive curves. PMID- 10676777 TI - Pediatric spinal injury. AB - Spine injury in children thankfully is still a relatively rare injury. The incidence of pediatric spine injuries has been reported as 2% to 5% of all spine injuries. The biological differences of children make differences in fracture patterns and alter the management necessary for successful treatment. The other factors that affect fracture production and associated injuries are head size relative to the body size, flexibility of the spine and supporting structures, the growth plates, and the elasticity and compressibility of the bone. The majority of compression injuries in children are made up of falls from a height. Spinal injuries in children remain a challenge despite some technical changes in assessment and treatment. PMID- 10676778 TI - Advances in the medical treatment of juvenile rheumatoid arthritis. AB - Juvenile rheumatoid arthritis (JRA) remains a challenge for clinicians. Nonsteroidal anti-inflammatory drugs and corticosteroids remain the mainstays of therapy, but concerns persist about side effects and the ability of these agents to prevent progression of bony disease. In recent years, novel treatments have been developed and quickly discarded because of unexpected toxicities or lack of efficacy. However, recent studies have shown that methotrexate and sulfasalazine are relatively safe and effective for JRA. Newly developed drugs, such as selective cyclooxygenase-2 inhibitors and soluble tumor necrosis factor receptor, whose development has stemmed from a more basic understanding of pathophysiology, may provide better disease control with fewer side effects. Finally, novel therapies, such as stem cell transplantation, may offer hope for children with JRA, especially systemic-onset JRA, whose disease has been refractory to conventional therapy. PMID- 10676780 TI - Bibliography. Current world literature. Hematology and oncology. PMID- 10676779 TI - Fever without apparent source on clinical examination, lower respiratory infections in children, and enterovirus infections. AB - This section focuses on issues in infectious disease that are commonly encountered in pediatric office practice. McCarthy discusses recent literature regarding the evaluation and management of acute fevers without apparent source on clinical examination in infants and children and the evaluation of children with prolonged fevers of unknown origin. Klig reviews recent literature about lower respiratory tract infection in children. Finally, Kennedy and Kahn discuss recent developments in infectious diseases pertinent to office practice. PMID- 10676781 TI - Bibliography. Current world literature. Orthopedics. PMID- 10676782 TI - Confirmation of confocal microscopy diagnosis of Acanthamoeba keratitis using polymerase chain reaction analysis. AB - BACKGROUND: Acanthamoeba keratitis has commonly been identified with in vivo confocal microscopy and confirmed with histologic examination of an epithelial biopsy specimen. OBJECTIVE: To determine if Acanthamoeba keratitis can be verified using polymerase chain reaction (PCR) of epithelial biopsy specimens. METHODS: Epithelial specimens from patients with suspected Acanthamoeba keratitis by confocal microscopy were tested for Acanthamoeba with PCR of Acanthamoeba ribosomal DNA. RESULTS: Twenty-four of 31 patients with evidence of Acanthamoeba keratitis were positive for Acanthamoeba on PCR analysis using 3 sets of primers. In 22 cases, the sequence obtained closely matched Acanthamoeba castellanii. CONCLUSIONS: This study demonstrates that PCR analysis of epithelial biopsy specimens can provide definitive verification of the confocal microscopic and histologic identification of Acanthamoeba organisms associated with keratitis. Acanthamoeba keratitis is probably quite common, especially in contact lens wearers, although more than half of the patients in this study did not wear contact lenses. PMID- 10676783 TI - Iridocorneal endothelial syndrome in Thai patients: clinical variations. AB - OBJECTIVE: To evaluate the spectrum of iridocorneal endothelial syndrome, to our knowledge, never studied previously in Orientals. METHODS: From 1986 to 1998, we examined 60 consecutive patients (20 men, 40 women) with characteristic signs of iridocorneal endothelial syndrome and compared the clinical manifestations to those reported in white patients. RESULTS: Cogan-Reese syndrome (CRS) was most common (38 patients), while 14 patients had Chandler syndrome (CS), and 8 had progressive iris atrophy. Three patients initially classified as having CS and 1 as having progressive iris atrophy progressed to CRS. Glaucoma occurred in 46 patients (76.7%), most commonly in patients with progressive iris atrophy or CRS. Ten patients had slow progression of disease during the follow-up period of up to 12 years. Three patients (2 with CRS, 1 with CS) had asymptomatic localized islands of "hammered-silver" appearance and 11 (8 with CRS, 2 with CS, and 1 with progressive iris atrophy) had subclinical abnormal endothelium in the contralateral eyes. A translucent membrane was commonly seen on the brown iris surface. Total endothelial involvement was present in 49 patients, while 6 (4 with CRS, 2 with CS) had focal endothelial abnormalities with sharp demarcation from adjacent normal endothelium. CONCLUSIONS: Iridocorneal endothelial syndrome occurs in Orientals. Cogan-Reese syndrome is the most common form and is strongly associated with glaucoma. Although several clinical manifestations were similar between whites and Orientals (mean age of onset, sex predilection, iris changes, peripheral anterior synechiae formation, or corneal edema), CRS was most prevalent; a translucent membrane were more noticeable in Orientals. PMID- 10676784 TI - Scanning laser ophthalmoscope correlations with biomicroscopic findings and foveal function after macular hole closure. AB - OBJECTIVE: To investigate the relation between foveal findings and visual function in eyes with a resolved idiopathic macular hole after vitreous surgery. METHODS: We divided 28 eyes with postoperative idiopathic macular hole resolution into 3 groups based on postoperative biomicroscopic foveal findings of complete closure, partial closure, or atrophic closure. To evaluate foveal retinal function, scanning laser ophthalmoscope (SLO) microperimetry was performed preoperatively and 6 months postoperatively. RESULTS: Postoperatively in 18 eyes (64%), the foveal images became normal or almost normal and were classified as having complete closure, 6 eyes (21%) were classified as having partial closure, and 4 eyes (14%) as having atrophic closure. The corresponding visual acuity levels 6 months postoperatively were, respectively, 0.10, 0.35, and 0.64 (P<.01) based on LogMAR analysis. Preoperative SLO microperimetry detected an absolute scotoma at the bottom of all macular holes; postoperatively, the absolute scotoma disappeared in the 18 eyes with complete hole closure, but a relative scotoma was detected in 6 eyes. Of 6 eyes with partial closure, 1 had an absolute scotoma and 5 had a relative scotoma. An absolute scotoma was detected in 4 eyes with atrophic closure. CONCLUSIONS: After macular hole closure, SLO findings correlate both with biomicroscopic findings and foveal function. Better anatomical foveal recovery in eyes after macular hole closure results in better improvement of vision than in eyes in which the foveal anatomical findings are not as good. PMID- 10676785 TI - Observations on 17 patients with retinocytoma. AB - OBJECTIVE: To study the clinical features and natural history of 17 patients with retinocytoma. DESIGN: A retrospective case series. SETTING: Tertiary referral center. PATIENTS: Data on 17 patients with retinocytoma were reviewed for clinical features. The natural history of retinocytoma and its risk for malignant transformation were also evaluated. RESULTS: Among 920 consecutive patients who had retinoblastoma, retinocytoma, or both, we identified 24 tumors in 17 patients (1.8%) with clinical features compatible with retinocytoma. The median age at diagnosis was 15 years (range, 4-45 years). Of the 24 tumors, the retinocytoma was bilateral in 3 cases (13%) and the family history of retinoblastoma was positive in 3 cases (13%). Seventeen (71%) of the tumors were extramacular in location, and 7 (29%) were located in the macular area. Ophthalmoscopic features characteristic of retinocytoma included the presence of a translucent retinal mass in 21 (88%), calcification in 15 (63%), and retinal pigment epithelial alteration in 13 (54%) of the 24 tumors. A combination of all 3 features was observed in 8 (33%) of the 24 tumors. In 13 (54%) of the tumors, a zone of chorioretinal atrophy could be observed. In 1 patient, subtle tumor regression was documented photographically. Only 1 retinocytoma (4%) underwent malignant transformation into retinoblastoma. At the last follow-up visit, none of the patients had developed a pineoblastoma or another second malignant neoplasm. CONCLUSIONS: Retinocytoma is a rare benign retinal tumor that has characteristic clinical features. The areas of chorioretinal atrophy were suggestive of tumor regression. In our series, the risk for malignant transformation of retinocytoma into retinoblastoma was 4%; therefore, patients with a presumed diagnosis of retinocytoma should be closely observed. PMID- 10676786 TI - Choroidal laser Doppler flowmetry in healthy subjects. AB - OBJECTIVE: To evaluate normal choroidal blood flow and its relationship with various factors such as age, systemic blood pressure, and intraocular pressure (IOP). METHODS: A total of 70 healthy subjects were recruited. Choroidal blood flow was assessed using a method based on laser Doppler flowmetry (LDF) technique. The LDF parameters of velocity, volume, and flux were obtained. The influence of age, mean systemic blood pressure, IOP, smoking, and sex on choroidal hemodynamic parameters was assessed in a multiple linear regression model. The correlations between interocular difference in IOP and interocular differences in the LDF parameters were assessed by means of the Pearson linear correlation factor. RESULTS: Velocity decreased significantly (P = .03) with advancing age of the subjects and volume increased significantly (P = .02) with increasing IOP. Mean blood pressure, smoking, and sex had no influence on the choroidal LDF parameters. Interocular difference in IOP correlated significantly with interocular difference in volume (R = 0.34, P<.005). CONCLUSION: Choroidal blood flow velocity decreased with increasing age of the subjects, while the volume of moving erythrocytes decreased with lower IOP. PMID- 10676787 TI - Metastatic neoplasms in the optic disc: the 1999 Bjerrum Lecture: part 2. AB - BACKGROUND: Little information is available on metastatic tumors to the optic disc. OBJECTIVE: To determine the clinical features and prognosis of patients with optic disc metastasis. DESIGN: Retrospective chart review. RESULTS: Of 660 consecutively evaluated patients with intraocular metastasis, 30 (4.5%) (31 eyes) had metastatic cancer to the optic disc; 24 (80%) were women and 6 (20%) were men. Mean age at the time of ocular diagnosis was 55 years. The primary neoplasm was in the breast in 13 patients (43%), in the lung in 8 (27%), in the intestine in 1 (3%), in the kidney in 1 (3%), and in the prostate in 1 (3%); the primary neoplasm was never determined in 6 patients (20%). The optic disc metastasis was unilateral in 29 patients (97%) and bilateral in 1 (3%). Ophthalmoscopically, the disc metastasis appeared as a diffuse enlargement of the optic disc in 26 eyes (84%) and as a distinct nodule in 5 (16%). There was an adjacent juxtapapillary choroidal component to the metastatic disc lesion in 23 eyes (74%), and the optic disc was involved without a retinal or choroidal component in 8 (26%). Other associated findings included some degree of secondary disc edema in all eyes, buried disc blood vessels in 23 (74%), and splinter hemorrhages in 13 (42%). Fine needle aspiration biopsy was useful in establishing the diagnosis in all 5 eyes in which it was performed. Mean survival was 13 months after diagnosis of the disc metastasis. CONCLUSIONS: Metastasis to the optic disc accounts for 5% of all intraocular metastases. It can occur as invasion from a juxtapapillary choroidal metastasis or as isolated optic disc metastasis. Breast and lung cancers are the most common primary neoplasms that account for metastasis to the optic disc. The primary site is never determined in 20% of patients. The characteristic clinical features of optic disc metastasis should help differentiate it from other causes of swollen optic disc. Patient prognosis is poor. PMID- 10676788 TI - Melkersson-Rosenthal syndrome: new clinicopathologic findings in 4 cases. AB - OBJECTIVE: To define the clinicopathologic features of eyelid involvement in Melkersson-Rosenthal syndrome (MRS). METHODS: Four patients with eyelid edema consistent with MRS were evaluated clinically, including diagnostic imaging in 2 patients. Eyelid tissue from these patients was examined by light microscopy and immunohistochemistry. Polymerase chain reaction for herpes simplex virus was performed in 1 case. RESULTS: The 3 men and 1 woman ranged in age from 33 to 74 years. All patients had insidious, painless, nonpitting eyelid edema. Three patients had unilateral edema; one had bilateral, asymmetric involvement. Ipsilateral lip edema was present in 1 case. Computed tomography demonstrated periorbital heterogeneous thickening that corresponded to the microscopic finding of scattered granulomas. All 4 patients demonstrated epithelioid granulomas inside and adjacent to dilated lymphatic vessels. Polymerase chain reaction testing was negative for herpes simplex virus. CONCLUSIONS: Isolated eyelid swelling that mimics thyroid-associated ophthalmopathy may occur in MRS. Computed tomography may be useful in the diagnosis. Biopsy should be performed in all cases of unexplained nonpitting eyelid edema. In the eyelid, MRS is characterized histopathologically by a granulomatous lymphangitis, a finding that seems to be unique to this condition. PMID- 10676789 TI - Swelling and loss of photoreceptors in chronic human and experimental glaucomas. AB - OBJECTIVE: To determine whether outer retinal changes occur in chronic, presumed primary open-angle glaucoma (POAG). METHODS: The outer retinas from 128 human eyes with a diagnosis of chronic glaucoma (presumably POAG in most cases) and 90 control eyes were examined histologically by 3 masked observers for photoreceptor swelling and loss. Retinas from 9 rhesus monkeys with glaucoma induced experimentally by laser trabecular destruction were compared with 7 fellow (control) eyes. The mean pressure elevations in the eyes with laser trabecular destruction ranged from 26.6 to 53.6 mm Hg with durations varying from 7 to 33 weeks. RESULTS: Swelling of the red- and green-sensitive cones was observed in a statistically significantly greater proportion of human eyes with presumed POAG compared with the control eyes. Patchy loss of red/green cones and rods was also found in some of the glaucomatous retinas. In a subset of the human eyes with end stage disease, cone swelling was a variable finding. Although no photoreceptor loss was found in the 9 monkey eyes with experimental glaucoma, 8 had swelling of their red/green cones that was remarkably similar to that seen in the human eyes. Swelling was not present in any of the control monkey eyes. CONCLUSIONS: The photoreceptors are affected by chronically elevated intraocular pressure. CLINICAL RELEVANCE: These findings may explain some of the abnormalities of color vision and the electrophysiological effects that have been observed in patients with POAG. PMID- 10676790 TI - Tear production after unilateral removal of the main lacrimal gland in squirrel monkeys. AB - OBJECTIVE: To study the effects of lacrimal gland removal on basal and reflex tear production and on the ocular surface in the squirrel monkey. METHODS: Unilateral main lacrimal gland removal in 6 squirrel monkeys was followed by Schirmer testing, slit-lamp examination with fluorescein, and collection of basal and reflex (stimulated) tears for analysis of tear protein spectra between 0 and 20 kd, as well as histological evaluation. RESULTS: Schirmer test results showed an 80% decrease in basal tears and a 90% decrease in reflex tears during week 1, and a 32.2% and 33.3% decrease, respectively, at week 20 after surgery, compared with the contralateral control side. However, no gross abnormalities or fluorescein staining were seen in 5 of the 6 monkeys, and the conjunctival surfaces remained normal. The main and accessory lacrimal glands appeared to secrete similar types of proteins. No histological changes were seen in corneal, conjunctival, or eyelid tissues 20 weeks after surgery. CONCLUSIONS: Tears from accessory lacrimal glands were sufficient to maintain a stable tear layer on the cornea, suggesting that so-called basal tear flow is made up of fluid from both main and accessory lacrimal glands and that decreased tear production by the main lacrimal gland is not a causative factor in keratoconjunctivitis sicca. CLINICAL RELEVANCE: This study shows that total removal of the main lacrimal gland does not in itself lead to keratoconjunctivitis sicca. However, the nature of neural control of the accessory glands is not yet clear. PMID- 10676791 TI - Surgical undertreatment of glaucoma in black beneficiaries of medicare. AB - OBJECTIVE: To identify whether there was surgical undertreatment of glaucoma in black beneficiaries of Medicare from 1991 to 1994. METHODS: We performed a retrospective cohort analysis on all argon laser trabeculoplasty or trabeculectomy surgery claims to the Health Care Financing Administration between 1991 and 1994. There were 191 287 Medicare patients who were black or white, at least 65 years of age, and resided in the United States at the time of their glaucoma surgery. Age- and sex-adjusted rates were obtained and compared with surgery rates expected based on disease prevalence. RESULTS: The age-sex-adjusted rate ratio of glaucoma surgical procedures for blacks to whites was 2.14. Assuming that treatments should be done in proportion to age-race prevalence, blacks undergo glaucoma surgery at approximately 47% below the expected rate. CONCLUSIONS: Blacks underwent argon laser trabeculoplasties and trabeculectomies at half the rate of whites from 1991 to 1994. Although in 1993 and 1994 there was a slight trend toward higher surgery rates in blacks, the magnitude of this improvement was small compared with estimated differences in the surgery rates between blacks and whites. PMID- 10676792 TI - Anterior chamber depth measurement as a screening tool for primary angle-closure glaucoma in an East Asian population. AB - OBJECTIVE: To evaluate anterior chamber depth measurement as a method of screening for primary angle-closure glaucoma in an East Asian population. DESIGN: Two-phase, cross-sectional, community-based study. SETTING: Rural and urban locations in the Hovsgol and Omnogobi provinces, Mongolia. PARTICIPANTS: Nine hundred forty-two (94.2%) of 1000 individuals in Hovsgol(1995) and 775 (96.9%) of 1000 individuals in Omnogobi (1997) aged 40 years or older were examined. MAIN OUTCOME MEASURES: Anterior chamber depth was measured by optical pachymetry, slitlamp-mounted A-mode ultrasound, and handheld ultrasound. Gonioscopy was used to detect occludable angles, defined as one in which the trabecular meshwork was visible for less than 90 degrees of angle circumference. Primary open-angle glaucoma was diagnosed in subjects with an occludable angle and glaucomatous optic neuropathy with visual morbidity. The area under the curve in a receiver operating characteristic plot was used to compare test performance. RESULTS: Optical pachymetry outperformed the slitlamp-mounted ultrasound method of anterior chamber depth measurement (area under the curve, 0.93 and 0.90, respectively; z test, P = .001). Handheld ultrasound (area under the curve, 0.86) was inferior to optical measurement (z test, P = .001) but did not differ significantly from slitlamp ultrasound (z test, P = .06). The optical method gave sensitivity of 85% and specificity of 84% at a screening cutoff of less than 2.22 mm for detecting occludable angles. CONCLUSIONS: Measurement of axial anterior chamber depth can detect occludable angles in this Asian population and therefore may have a role in population screening for primary angle-closure glaucoma. PMID- 10676793 TI - Age-specific causes of bilateral visual impairment. AB - OBJECTIVES: To describe the age-specific prevalence of common eye diseases causing bilateral visual impairment and estimate the total number of Australians with cause-specific visual impairment. METHODS: Cluster-stratified random sample of 5147 residents aged 40 years and older from urban and rural areas and aged care facilities. Participants completed a standardized interview and eye examination. Four levels of bilateral visual impairment were defined: less than 20/40 to 20/60 and/or homonymous hemianopia (mild), less than 20/60 to 20/200 or better and/or less than 20 degrees to 10 degrees radius field (moderate), less than 20/200 to 10/200 and/or less than 10 degrees to 5 degrees radius field (severe), and less than 1O/ 200 and/or less than 50 radius field (profound). The major cause of vision loss was identified for all participants found to be visually impaired. RESULTS: Uncorrected refractive error was the most common cause of bilateral visual impairment across all decades of life, rising from 0.5% in 40- to 49-year-olds to 13% among those aged 80 years and older. Prevalence of visual impairment due to diabetic retinopathy was 0.7% in 50- to 59-year-olds and 0.8% in those older than 80 years. Visual impairment due to glaucoma had a prevalence of 0.7% among 60-year-olds and rose to 4% of those older than 90 years. The prevalence of visual impairment due to cataract (only present in those aged 70 years or older) rose from 0.6% to 11% in those older than 90 years, and the prevalence of visual impairment due to age-related macular degeneration rose from 0.8% to 16% in those older than 90 years. CONCLUSIONS: The predominant causes of visual impairment change with age. Recognition of these patterns is fundamental for early diagnosis and treatment of eye disease and, where appropriate, referral for rehabilitation. PMID- 10676794 TI - The analysis of clinical research: mandatory submission of data sets. Journals should have access to research data. PMID- 10676795 TI - Journal editors should not require complete data sets for independent scrutiny. PMID- 10676796 TI - Management of small corneal infiltrates in contact lens wearers. PMID- 10676797 TI - T-cell lymphoproliferative disorder of vitreous associated with mycosis fungoides. AB - 59-year-old man with a history of mycosis fungoides developed loss of visual acuity and visual field in the left eye. Epiretinal lesions were present in the right eye and multifocal choroidal lesions, optic disc edema, and vitritis were present in the left eye. A diagnostic vitrectomy was performed and cytologic examination of the vitreous confirmed the diagnosis of T-cell lymphoproliferative disorder. Systemic and intrathecal chemotherapy resulted in marked improvement in ocular signs and symptoms. At last follow-up, the patient was found to have improved visual acuity in the left eye; however, significant worsening of his systemic condition developed and he died shortly thereafter. PMID- 10676798 TI - Photorefractive keratectomy for correction of epikeratophakia regression. PMID- 10676799 TI - Iatrogenic keratoconus: corneal ectasia following laser in situ keratomileusis for myopia. PMID- 10676800 TI - Treatment of conjunctival mucosa-associated lymphoid tissue lymphoma with intralesional injection of interferon alfa-2b. PMID- 10676801 TI - Spontaneous resolution of vitreomacular traction documented by optical coherence tomography. PMID- 10676802 TI - Acute bilateral visual loss associated with retinal hemorrhages following epiduroscopy. PMID- 10676803 TI - Capillary hemangioma of the optic nerve head and juxtapapillary retina. PMID- 10676804 TI - Sildenafil (Viagra) associated anterior ischemic optic neuropathy. PMID- 10676805 TI - Mycobacterium chelonae infection in a corneal graft. PMID- 10676806 TI - Peculiar macular hemorrhage. PMID- 10676807 TI - Retinopathy of NARP syndrome. PMID- 10676808 TI - Codons 837 and 838 in the retinal guanylate cyclase gene on chromosome 17p: hot spots for mutations in autosomal dominant cone-rod dystrophy? PMID- 10676809 TI - Yeast prions and their prion-forming domain. PMID- 10676810 TI - New roles for Src kinases in control of cell survival and angiogenesis. PMID- 10676811 TI - Time flies for Drosophila. PMID- 10676812 TI - Large-scale movement of elongation factor G and extensive conformational change of the ribosome during translocation. AB - Elongation factor (EF) G promotes tRNA translocation on the ribosome. We present three-dimensional reconstructions, obtained by cryo-electron microscopy, of EF-G ribosome complexes before and after translocation. In the pretranslocation state, domain 1 of EF-G interacts with the L7/12 stalk on the 50S subunit, while domain 4 contacts the shoulder of the 30S subunit in the region where protein S4 is located. During translocation, EF-G experiences an extensive reorientation, such that, after translocation, domain 4 reaches into the decoding center. The factor assumes different conformations before and after translocation. The structure of the ribosome is changed substantially in the pretranslocation state, in particular at the head-to-body junction in the 30S subunit, suggesting a possible mechanism of translocation. PMID- 10676813 TI - The crystal structure of human eukaryotic release factor eRF1--mechanism of stop codon recognition and peptidyl-tRNA hydrolysis. AB - The release factor eRF1 terminates protein biosynthesis by recognizing stop codons at the A site of the ribosome and stimulating peptidyl-tRNA bond hydrolysis at the peptidyl transferase center. The crystal structure of human eRF1 to 2.8 A resolution, combined with mutagenesis analyses of the universal GGQ motif, reveals the molecular mechanism of release factor activity. The overall shape and dimensions of eRF1 resemble a tRNA molecule with domains 1, 2, and 3 of eRF1 corresponding to the anticodon loop, aminoacyl acceptor stem, and T stem of a tRNA molecule, respectively. The position of the essential GGQ motif at an exposed tip of domain 2 suggests that the Gln residue coordinates a water molecule to mediate the hydrolytic activity at the peptidyl transferase center. A conserved groove on domain 1, 80 A from the GGQ motif, is proposed to form the codon recognition site. PMID- 10676814 TI - Sequence-specific RNA binding by a Nova KH domain: implications for paraneoplastic disease and the fragile X syndrome. AB - The structure of a Nova protein K homology (KH) domain recognizing single stranded RNA has been determined at 2.4 A resolution. Mammalian Nova antigens (1 and 2) constitute an important family of regulators of RNA metabolism in neurons, first identified using sera from cancer patients with the autoimmune disorder paraneoplastic opsoclonus-myoclonus ataxia (POMA). The structure of the third KH domain (KH3) of Nova-2 bound to a stem loop RNA resembles a molecular vise, with 5'-Ura-Cyt-Ade-Cyt-3' pinioned between an invariant Gly-X-X-Gly motif and the variable loop. Tetranucleotide recognition is supported by an aliphatic alpha helix/beta sheet RNA-binding platform, which mimics 5'-Ura-Gua-3' by making Watson-Crick-like hydrogen bonds with 5'-Cyt-Ade-3'. Sequence conservation suggests that fragile X mental retardation results from perturbation of RNA binding by the FMR1 protein. PMID- 10676815 TI - Role of Sec61alpha in the regulated transfer of the ribosome-nascent chain complex from the signal recognition particle to the translocation channel. AB - Targeting of ribosome-nascent chain complexes to the translocon in the endoplasmic reticulum is mediated by the concerted action of the signal recognition particle (SRP) and the SRP receptor (SR). Ribosome-stripped microsomes were digested with proteases to sever cytoplasmic domains of SRalpha, SRbeta, TRAM, and the Sec61 complex. We characterized protein translocation intermediates that accumulate when Sec61alpha or SRbeta is inactivated by proteolysis. In the absence of a functional Sec61 complex, dissociation of SRP54 from the signal sequence is blocked. Experiments using SR proteoliposomes confirmed the assembly of a membrane-bound posttargeting intermediate. These results strongly suggest that the Sec61 complex regulates the GTP hydrolysis cycle of the SRP-SR complex at the stage of signal sequence dissociation from SRP54. PMID- 10676816 TI - Structure of the Rho family GTP-binding protein Cdc42 in complex with the multifunctional regulator RhoGDI. AB - The RhoGDI proteins serve as key multifunctional regulators of Rho family GTP binding proteins. The 2.6 A X-ray crystallographic structure of the Cdc42/RhoGDI complex reveals two important sites of interaction between GDI and Cdc42. First, the amino-terminal regulatory arm of the GDI binds to the switch I and II domains of Cdc42 leading to the inhibition of both GDP dissociation and GTP hydrolysis. Second, the geranylgeranyl moiety of Cdc42 inserts into a hydrophobic pocket within the immunoglobulin-like domain of the GDI molecule leading to membrane release. The structural data demonstrate how GDIs serve as negative regulators of small GTP-binding proteins and how the isoprenoid moiety is utilized in this critical regulatory interaction. PMID- 10676817 TI - Covalent modification regulates ligand binding to receptor complexes in the chemosensory system of Escherichia coli. AB - In the Escherichia coli chemosensory pathway, receptor modification mediates adaptation to ligand. Evidence is presented that covalent modification influences ligand binding to receptors in complexes with CheW and the kinase CheA. Kinase inhibition was measured with serine receptor complexes in different modification levels; Ki for serine-mediated inhibition increased 10,000-fold from the lowest to the highest level. Without CheA and CheW, ligand binding is unaffected by covalent modification; thus, the influence of covalent modification is mediated only in the receptor complex, a conclusion supported by an analogy to allosteric enzymes and the observation of cooperative kinase inhibition. Also, the finding that a subsaturating serine concentration accelerates active receptor-kinase complex assembly implies that the assembly/disassembly process may also contribute to kinase regulation. PMID- 10676818 TI - Integrated cytogenetic map of chromosome arm 4S of A. thaliana: structural organization of heterochromatic knob and centromere region. AB - We have constructed an integrated cytogenetic map of chromosome arm 4S of Arabidopsis thaliana. The map shows the detailed positions of various multicopy and unique sequences relative to euchromatin and heterochromatin segments. A quantitative analysis of the map positions at subsequent meiotic stages revealed a striking pattern of spatial and temporal variation in chromatin condensation for euchromatin and heterochromatin. For example, the centromere region consists of three domains with distinguishable structural, molecular, and functional properties. We also characterized a conspicuous heterochromatic knob of approximately 700 kb that accommodates a tandem repeat and several dispersed pericentromere-specific repeats. Moreover, our data provide evidence for an inversion event that relocated pericentromeric sequences to an interstitial position, resulting in the heterochromatic knob. PMID- 10676819 TI - The complete sequence of a heterochromatic island from a higher eukaryote. The Cold Spring Harbor Laboratory, Washington University Genome Sequencing Center, and PE Biosystems Arabidopsis Sequencing Consortium. AB - Heterochromatin, constitutively condensed chromosomal material, is widespread among eukaryotes but incompletely characterized at the nucleotide level. We have sequenced and analyzed 2.1 megabases (Mb) of Arabidopsis thaliana chromosome 4 that includes 0.5-0.7 Mb of isolated heterochromatin that resembles the chromosomal knobs described by Barbara McClintock in maize. This isolated region has a low density of expressed genes, low levels of recombination and a low incidence of genetrap insertion. Satellite repeats were absent, but tandem arrays of long repeats and many transposons were found. Methylation of these sequences was dependent on chromatin remodeling. Clustered repeats were associated with condensed chromosomal domains elsewhere. The complete sequence of a heterochromatic island provides an opportunity to study sequence determinants of chromosome condensation. PMID- 10676820 TI - Efficacy and safety of lesopitron in outpatients with generalized anxiety disorder. AB - OBJECTIVE: To compare the relative efficacy and safety of lesopitron 4-80 mg/d versus lorazepam 2-4 mg/d and placebo in a subgroup of patients with anxiety history taken from a larger study of patients with a primary diagnosis of generalized anxiety disorder (GAD). DESIGN: Six-week, randomized, double-blind, parallel, placebo and lorazepam-controlled, Phase II, single-center, outpatient study. SETTING: Outpatient clinic. PATIENTS: One hundred sixty-one patients with GAD were randomized in the main study; 68 with a documented history of GAD or anxiety disorder not otherwise specified were included in the subgroup. METHODS: After a one-week placebo lead-in, patients were randomized to receive placebo, lesopitron, or lorazepam twice daily for six weeks, followed by a one-week taper period. Efficacy was assessed using the Hamilton Rating Scale for Anxiety (HAM-A) and the Clinical Global Impressions scale. Safety was assessed through physical examinations, monitoring of vital signs, 12-lead electrocardiograms, laboratory analyses, and adverse event monitoring. RESULTS: An overall mean improvement in the HAM-A total score between baseline and end point for all three treatment groups was seen, with mean changes of 3.4 (95% CI 2.0 to 4.8), 6.1 (95% CI 4.1 to 8.1), and 6.1 (95% CI 4.6 to 7.6) for the placebo, lesopitron, and lorazepam groups, respectively (omnibus p = 0.044, uncorrected). Positive treatment effects were also observed in the subgroup population on several other measures and suggest that additional therapeutic trials may be warranted. Future trials could be stratified on the basis of referral status (symptomatic volunteer vs. clinical patient with preexisting illness) or previous exposure to anxiolytics, and use a fixed-dose rather than flexible-fixed-dose design. CONCLUSIONS: The subgroup analysis represents a comparison of treatment outcome in GAD patients presenting with a history of previous episodes of GAD or anxiety disorder not otherwise specified compared with those who were experiencing their first episode of GAD and reported no anxiety history. Although the overall study analysis was equivocal, for the approximately 40% of patients with recurrent anxiety disorder, beneficial effects for both lesopitron and lorazepam are suggested. PMID- 10676821 TI - Economic impact of standardized orders for antimicrobial prophylaxis program. AB - OBJECTIVE: To assess the effect and economic impact of an intervention aimed at standardizing the timing of preoperative antimicrobial prophylaxis from the perspective of a major teaching hospital. DESIGN: A pre/post study design in which a random sample of 60 procedures from a 12-month period in the preintervention phase were reviewed. A comparative sample of 60 procedures during a seven-month postintervention phase was selected. For each prophylactic course, preoperative dose administration details were classified as early (>2 h prior to incision), on time (0-2 h prior), delayed (0-3 h after), or late (>3 after). To determine the economic impact of this intervention, we used a predictive decision analytic model using institutional costs and the published probabilities of inpatient surgical wound infections (SWIs) following administration of antimicrobials timed according to the above criteria. Two conditions were analyzed: (1) an interdisciplinary two-stage therapeutic interchange program involving staff education and modification of preoperative antimicrobial orders to ensure timely administration and (2) no intervention. SETTING: An 1100-bed tertiary care, university-affiliated institution. PATIENTS: 120 randomly selected procedures involving inpatients who received a preoperative antibiotic. OUTCOME MEASURES: Differences in preoperative antimicrobial timing and cost avoidance associated with the intervention. RESULTS: In the preintervention phase, 68% of prophylactic courses were on time, 22% were early, and the balance were delayed or late. The incidence of on-time prophylaxis increased to 97% during the postintervention phase (p = 0.001). Operating room staff involvement in antimicrobial administration increased from 57% to 92% (p = 0.001). Based on a setup and annual intervention cost of $9100 CAN ($1 CAN = $0.68 US), an annual inpatient SWI avoidance of 51 cases, an average infection-associated extended hospital stay of four days, and an average treatment cost of $1957 CAN per inpatient SWI, we estimated that 153 hospital days were avoided and there was an annual cost avoidance of $90 707 CAN ($1779 CAN saved per inpatient infection avoided) due to this intervention. Using sensitivity analyses, no plausible changes in the base case estimates altered the results of the economic model. CONCLUSIONS: An interdisciplinary approach to optimizing the timing of preoperative antimicrobial doses can impact positively on practice patterns and result in substantial cost avoidance. Costs incurred to implement such an intervention are small when compared with the annual cost avoidance to the institution. PMID- 10676822 TI - The pharmacokinetics of etanercept in healthy volunteers. AB - OBJECTIVE: To describe the pharmacokinetics of etanercept when administered by subcutaneous injection in single doses to healthy volunteers. METHODS: Twenty-six healthy volunteers between 19 and 50 years of age received single doses of etanercept 25 mg by subcutaneous injection into the abdomen. Serial serum samples were collected for 21 days. An enzyme-linked immunosorbent assay with a quantitation limit of 0.3 ng/mL was used to measure the drug concentrations. RESULTS: Etanercept was well tolerated by healthy volunteers. A one-compartment model was found to best describe the concentration-time data and was used to determine the pharmacokinetic parameters. Etanercept is slowly absorbed from the site of injection with a time of peak concentration (+/- SD) of 51 +/- 14 hours; peak concentration was 1.46 +/- 0.72 mg/L. The AUC was 235 +/-98 mg x h/L, apparent clearance was 132 +/- 85 mL/h, apparent volume of distribution was 12 +/ 6 L, and the half-life was 68 +/- 19 hours. CONCLUSIONS: Etanercept was slowly absorbed and slowly eliminated after subcutaneous administration. Dosing at the recommended rate of 25 mg twice weekly would be expected to result in concentrations of approximately 3 mg/L. Intersubject variability for apparent clearance in healthy volunteers was 64%. PMID- 10676823 TI - Toxicity of polysaccharide--iron complex exposures reported to poison control centers. AB - OBJECTIVE: To evaluate the toxicity of polysaccharide-iron complex (PIC) exposures reported to poison centers in the US. DESIGN: A retrospective analysis of potentially toxic exposures to PIC without concomitant substances reported to the American Association of Poison Control Centers (AAPCC) Toxic Exposure Surveillance System from 1990 to 1998 was performed. RESULTS: Of 810 potentially toxic exposures to PIC, 55.9% occurred in females, 43.8% in males; in 0.3%, gender was unknown. The majority of exposures (74.4%) involved children under six years of age. The reasons for exposure were: 86.7% unintentional, 11.6% intentional, and 1.6% adverse reaction. The most frequently reported clinical effects attributed to PIC were vomiting (n = 23), diarrhea (10), nausea (11), abdominal pain (10), and lethargy/drowsiness (7). While the majority of exposures were managed outside a healthcare facility, management site varied depending on age (management in non-healthcare facility in 71.8% of exposures in children under six years of age vs. 44.9% in adolescents and adults). The majority of outcomes (95.6%) were no effect, minor effect, unrelated effect, not followed since nontoxic, or not followed since only minimal toxicity possible. Of two cases coded as moderate effect, it could not be determined whether the symptoms were related to PIC in one, and in the second case inspection of the poison center record revealed that the actual outcome was minor effect. There were no major effects or deaths. CONCLUSIONS: There were no serious adverse events following PIC exposure reported to the AAPCC. Although more data are needed, these findings suggest reduced toxicity for PIC relative to other forms of iron. PMID- 10676824 TI - Criteria for assessing the appropriateness of patient counseling in community pharmacies. AB - OBJECTIVE: To develop valid, reliable criteria for assessing the appropriateness of the management of common ailments and nonprescription drug therapy in community pharmacies in the UK. METHODS: The criteria were developed by an expert panel using the nominal group technique. The validity of the criteria was tested by surveying a random sample of pharmacists who were asked to rate the importance of each criterion on a semantic differential scale from 1 (low) to 7 (high). Subsequently, the reliability of the criteria was assessed: a random sample of pharmacists were each asked to apply the criteria to four vignettes of patient counseling on two separate occasions. RESULTS: All assessment criteria exceeded our predefined level of face, content, and consensual validity. In reliability testing, the overall assessment of appropriateness, along with five component assessment criteria, surpassed our predefined level of reliability. Three criteria, however, did not meet our predefined standard. These criteria were rational content of advice, rational product choice, and referral to another health professional. CONCLUSIONS: This represents the first systematic attempt to develop an instrument of general applicability for assessing the appropriateness of patient counseling and to subject it to rigorous validity and reliability testing. We suggest that further work is required to refine the criteria that did not meet reliability standards and to understand the decision-making processes underlying the assessment of vignettes of patient counseling. PMID- 10676825 TI - Syndrome of inappropriate antidiuretic hormone secretion associated with lisinopril. AB - OBJECTIVE: To describe a case of the syndrome of inappropriate antidiuretic hormone secretion (SIADH) associated with lisinopril therapy. CASE SUMMARY: A 76 year-old white woman who was being treated with lisinopril and metoprolol for hypertension presented with headaches accompanied by nausea and a tingling sensation in her arms. Her serum sodium was 109 mEq/L, with a serum osmolality of 225 mOsm/kg, urine osmolality of 414 mOsm/kg, and spot urine sodium of 122 mEq/L. Diclofenac 75 mg qd for osteoarthritic pain and lisinopril 10 mg qd for hypertension was begun in 1990. Lisinopril was increased to 20 mg qd in August 1994 and to 20 mg bid pm in August 1996 for increasing blood pressure; metoprolol 50 mg qd was added in July 1996. A diagnosis of SIADH was postulated and further evaluation was undertaken to exclude thyroid and adrenal causes. After lisinopril was discontinued and the patient restricted to 1000 mL/d of fluid, serum sodium gradually corrected to 143 mEq/L. The patient was discharged taking metoprolol alone for her hypertension; serum sodium has remained > or =138 mEq/L through April 1999, 32 months after discharge, despite daily use of diclofenac. DISCUSSION: Angiotensin-converting enzyme (ACE) inhibitors in antihypertensive doses may block conversion of angiotensin I to angiotensin II in the peripheral circulation, but not in the brain. Increased circulating angiotensin I enters the brain and is converted to angiotensin II, which may stimulate thirst and release of antidiuretic hormone from the hypothalamus, eventually leading to hyponatremia. CONCLUSIONS: SIADH should be considered a rare, but possible, complication of therapy with lisinopril and other ACE inhibitors. PMID- 10676826 TI - Malignant primary hypertension in pregnancy treated with lisinopril. AB - OBJECTIVE: To report a case of a patient treated with an angiotensin-converting enzyme (ACE) inhibitor with a good neonatal outcome. CASE REPORT: A 39-year-old African-Caribbean patient who had chronic hypertension presented at 18 weeks' gestation with acute hypertension. She was being treated for chronic hypertension with lisinopril, but had self-discontinued treatment. Attempts to control her hypertension with labetolol, nifedipine, and methyldopa were ineffective. She was therefore offered termination of pregnancy so treatment with lisinopril could be restarted. The patient elected to continue with the pregnancy in spite of the fetal risks associated with the use of an ACE inhibitor. She was delivered of a girl at 26 weeks' gestation. The baby initially had renal failure and also developed acute necrotizing enterocolitis. The renal failure improved simultaneously with the latter complication, and it is postulated that there was enteric excretion of lisinopril. The baby was discharged home on day 102 with no further complications. DISCUSSION: ACE inhibitors are acceptable medications to use in the first trimester of pregnancy; however, fetal malformations and neonatal complications have been associated with their use later in pregnancy, and they have a perinatal mortality rate of 97/1000. Lisinopril is excreted in urine and feces unchanged, and its half-life is prolonged in anuric neonates. Peritoneal dialysis eliminates lisinopril; however, this neonate improved after treatment for necrotizing enterocolitis and simultaneous improvement in bowel function. CONCLUSIONS: ACE inhibitors should not be used in pregnancy beyond the end of the first trimester. In exceptional cases, they may be indicated for the control of severe hypertension when the patient is refractory to other medications. The patient should be fully counseled about the adverse effect profile and neonatal outcome. This case report documents a successful outcome for mother and baby in these circumstances. PMID- 10676827 TI - Elevated free fractions of valproic acid in a heart transplant patient with hypoalbuminemia. AB - OBJECTIVE: To report a case demonstrating the importance of monitoring unbound valproic acid (VPA) serum concentrations in a patient with hypoalbuminemia. CASE SUMMARY: A 53-year-old white woman status-post heart transplantation was admitted to the hospital for declining cardiac function, possible rejection, and increased lethargy requiring intubation. An extensive workup of the patient's profound lethargy was initiated, including an evaluation of her VPA regimen. Initially, VPA dosages were adjusted based on the total serum concentration of VPA. Hypoalbuminemia compounded with increased lethargy prompted the measurement of unbound serum concentrations of VPA. The VPA dosage was then adjusted based on the unbound rather than the total VPA serum concentration; the patient eventually improved and was discharged from the hospital. DISCUSSION: Lethargy is a concentration-related adverse effect of VPA. The nonlinear pharmacokinetic and protein saturation characteristics of VPA may result in nonproportional elevations in unbound drug, and subsequent increases in adverse effects, when dosage adjustments are based solely on measurement of total VPA serum concentrations in patients with hypoalbuminemia. CONCLUSIONS: This case report suggests that appropriate monitoring of unbound drug concentrations of VPA may prevent unrecognized concentration-related adverse effects. Awareness of the pharmacokinetic relationship and adverse effects of VPA will aid clinicians in identifying the etiology of symptoms. PMID- 10676828 TI - Tamsulosin for the treatment of benign prostatic hypertrophy. AB - OBJECTIVE: To review the information necessary to assess the efficacy and safety of tamsulosin compared with other adrenergic antagonists for treatment of symptomatic benign prostatic hyperplasia. DATA SOURCES: A search was conducted of Cumulated Index Medicus, January 1993-August 1999, which was restricted to human trials and English-language journals. STUDY SELECTION AND DATA EXTRACTION: Efficacy studies were included if the design was randomized and included a control group. Drug safety was assessed using data from any patient series or controlled study. DATA SYNTHESIS: Tamsulosin, a uroselective alpha1A-adrenergic receptor antagonist, relaxes smooth muscle in the prostate and bladder neck, thereby enhancing bladder emptying. In randomized, controlled clinical trials using standardized instruments, tamsulosin improves obstructive voiding symptoms by at least 25% in 65-80% of patients with symptomatic benign prostatic hyperplasia. Tamsulosin also improves peak urinary flow rate by 1.4-3.6 mL/sec in various studies and reduces post-void residual urine volume. The usual dosage of tamsulosin was 0.4 or 0.8 mg orally once a day in the studies performed in the US and Europe; daily doses of 0.1-0.4 mg were used in studies performed in Japan. The beneficial effects of tamsulosin on voiding symptoms, peak urinary flow rate, and bladder emptying appear to be dose-related, up to a ceiling dose of 0.4 mg. The most common adverse effects are headache, asthenia, dizziness, and rhinitis like complaints. Retrograde or delayed ejaculation occurs in 4.5-14.0% of patients and has required discontinuation of treatment in a minority of these patients. At the usual dose of 0.4-0.8 mg/d, tamsulosin does not appear to significantly reduce blood pressure, increase heart rate, or cause first-dose syncope; therefore, dosage titration is not necessary when initiating treatment. Use of nifedipine, enalapril, atenolol, furosemide, or digoxin does not require dosage modification when tamsulosin is initiated concomitantly; hypotension has not been reported with combined use of tamsulosin and these commonly used agents. CONCLUSIONS: Tamsulosin is an improvement over other alpha-adrenergic antagonists for the management of symptoms of benign prostatic hyperplasia. It is a more convenient alternative that does not require initial dosage titration, has a fast onset of action, and has a low potential to cause hypotension when used alone or in combination with commonly used antihypertensive agents. It is more costly than some of the other second-generation alpha-adrenergic antagonists. PMID- 10676829 TI - Switching antipsychotic therapies. AB - BACKGROUND: Atypical antipsychotics are superior to conventional antipsychotics in improving positive and negative psychotic symptoms. Atypical antipsychotics do not exacerbate mood symptoms, and may improve mood symptoms and cognitive functioning; additionally, they have better adverse effect profiles than conventional antipsychotics. OBJECTIVE: To review the benefits of switching patients with schizophrenia or schizoaffective disorder from a conventional to an atypical antipsychotic, or from one atypical antipsychotic to another. In spite of the higher acquisition cost of atypical antipsychotics, overall treatment costs may decrease due to lower relapse and hospitalization rates. DATA SOURCES: A MEDLINE search (January 1977-January 1999) was conducted for articles written in English about efficacy, adverse effects, compliance, and pharmacoeconomics for atypical and conventional antipsychotics. STUDY SELECTION: Large, multicenter, double-blind, controlled studies were used for efficacy, safety, tolerability, and pharmaco-economic data. Where appropriate, recent review articles were also used. RESULTS: Atypical antipsychotics are superior to conventional antipsychotics in the treatment of schizophrenia. Atypical and conventional antipsychotics have different adverse effect profiles, costs, and compliance rates. CONCLUSIONS: Some patients may benefit by switching from a conventional to an atypical antipsychotic, from an atypical to a conventional antipsychotic, or from one atypical antipsychotic to another. Methods of switching antipsychotic therapies include tapering and cross-over strategies. PMID- 10676830 TI - Newer antithrombotic strategies in the initial management of non-ST-segment elevation acute coronary syndromes. AB - OBJECTIVE: To review the place in therapy of currently available antithrombotic agents in the non-ST-segment elevation acute coronary syndromes, that is, unstable angina and non-Q-wave myocardial infarction (MI). Recommendations are made based on currently available data. DATA SOURCE: English-language clinical studies, position statements, and review articles pertaining to the management of unstable angina and non-Q-wave MI with currently available products. STUDY SELECTION: Selection of prospective clinical studies was limited to those focusing on the management of the non-ST-segment elevation acute coronary syndromes, unstable angina, and non-Q-wave MI. DATA SYNTHESIS: It has yet to be determined which combination of agents (dalteparin, enoxaparin, lepirudin, clopidogrel, ticlopidine, abciximab, eptifibatide, tirofiban) and procedural strategies most significantly reduces mortality and serious events in these patients. The relevant pathophysiology, diagnostic criteria, and risk-stratifying procedures are reviewed in context with information from clinical studies regarding currently available agents for the management of non-ST-segment elevation acute coronary syndromes. CONCLUSIONS: A large number of new therapeutic classes and agents are available for the treatment of unstable angina and non-Q-wave MI. Although the diagnoses of unstable angina or non-Q-wave MI identify risk, treatment decisions are often based on the presence or absence of ST-segment elevations. Limited prospective evidence delineates the proper utilization of resources to best manage these patients. Efforts should be aimed at identifying particular patients who will best benefit from recently available therapies. PMID- 10676831 TI - Immunology of Varicella Immunization in the elderly. AB - OBJECTIVE: To review the varicella-zoster virus (VZV) and herpes zoster disease and to summarize published reports on the use of the live-attenuated varicella zoster vaccine to enhance cell-mediated immunity in elderly individuals. DATA SOURCE: A MEDLINE search (1966-August 1999) for English-language clinical studies and review articles pertaining to VZV and the live-attenuated varicella vaccine was conducted; references obtained from these publications were subsequently reviewed for additional relevant articles. STUDY SELECTION AND DATA EXTRACTION: Representative clinical trials were summarized and relevant information was selected to assist in the understanding of VZV, the subsequent immune response, and the live-attenuated varicella vaccine. DATA SYNTHESIS: The physiologic, age related decline in VZV cell-mediated immunity has been shown to be restored on administration of live-attenuated varicella vaccine. Various studies report serum anti-VZV antibody concentrations, and production of interferon-gamma were increased following vaccination. Concentrations subsequently returned to baseline one year after vaccination. Increase in responder cell frequency, a measure of cell-mediated immunity, has been reported to last up to four years after vaccination, at concentrations similar or superior to those observed following herpes zoster. CONCLUSIONS: Enhancement of cell-mediated immune response in elderly individuals through vaccination with live-attenuated varicella vaccine is a possible measure to protect this population from herpes zoster and to attenuate its complications. A summary of immunogenicity studies to identify the immune response to live-attenuated varicella vaccine in the elderly is presented. The absolute clinical significance, as well as appropriate administration guidelines of this prophylactic intervention, will become evident following forthcoming large, masked, placebo-controlled trials. PMID- 10676832 TI - Anterior ocular infections: an overview of pathophysiology and treatment. AB - OBJECTIVE: To provide a review of the pathophysiology and treatment of anterior ocular infections. DATA SOURCE: A MEDLINE search (from 1970 to October 1998) as well as a review of the tertiary literature was performed to identify pertinent literature on pathophysiology and treatment of ocular infections. STUDY SELECTION AND DATA EXTRACTION: All articles were considered for possible inclusion in the review. Relevant studies were selected for discussion in the article. DATA SYNTHESIS: Ocular infections are common and vary from self-limiting to sight threatening. Infections occur in different eye structures; presentation and treatment vary accordingly. Infections can occur when tissues of the eye are exposed to pathogens not normally present, when the eye is damaged, allowing pathogens to overcome the natural defenses of the eye, or in immunosuppressed patients where normal flora may become opportunistic. In deciding on appropriate treatment, both the causative pathogen and the structure(s) affected must be considered. The most likely pathogen can often be determined based on clinical signs and symptoms, patient history, or, in some cases, may need to be determined microbiologically. Differences in drug absorption, penetration, and availability to the various structures of the eye affect treatment decisions. Severity of infection, efficacy and safety of medication, and cost/benefit ratios must be taken into consideration in choosing the proper pharmacologic management of various ocular infections. CONCLUSIONS: Treatment of ocular infections depends on knowledge of the pathophysiology and drug disposition at the site of infection. An understanding of the current concepts surrounding the management of the anterior ocular infections presented will aid in the provision of optimal patient care. PMID- 10676833 TI - Management protocol for abacavir-related hypersensitivity reaction. AB - OBJECTIVE: To develop an abacavir hypersensitivity reaction management protocol for the University of Oklahoma Health Sciences Center. DATA SYNTHESIS: Conference abstracts, published literature, and manufacturer-provided materials were reviewed to define the syndrome and manifestations and to recommend steps to take when a patient develops abacavir-related reactions. RESULTS: Initial education, formalized contact and response mechanism, and intervention levels were incorporated into a protocol for clinicians. CONCLUSIONS: Abacavir, a newly approved agent for the treatment of HIV, has been associated with a fatal hypersensitivity reaction. Our protocol provides a mechanism to minimize progression of abacavir hypersensitivity reaction by providing a formalized management procedure. PMID- 10676834 TI - The role of vasopressin in the treatment of vasodilation in shock states. AB - OBJECTIVE: To review the role of vasopressin in the treatment of vasodilatory shock. DATA SOURCES: A MEDLINE search on published reports (1966-April 1999) was conducted. STUDY SELECTION: English-language studies and case reports were selected and evaluated based on quality of review of vasopressin in the treatment of vasodilatory shock. DATA SYNTHESIS: In patients with end-stage vasodilatory shock, baroreceptor reflex is impaired and vasopressin stores are depleted. Persistent elevation of catecholamines may lead to down-regulation of beta adrenergic receptors and reduces smooth-muscle response to catecholamines, leading to inability of maintaining organ perfusion. Small-scale studies and case reports have demonstrated vasopressin's efficacy in maintaining blood pressure in patients with septic shock, cardiac arrest, and end-stage heart failure, refractory to other vasopressor therapies. CONCLUSIONS: Vasopressin may be a reasonable alternative for patients in vasodilatory shock. However, larger-scale controlled dinical trials are warranted before its routine use can be recommended. PMID- 10676835 TI - Do ethanol and metronidazole interact to produce a disulfiram-like reaction? AB - OBJECTIVE: To obtain and evaluate evidence about the supposed disulfiram-like interaction between metronidazole and ethanol. DATA SOURCES: MEDLINE search from January 1964 to June 1999, using the terms metronidazole, ethanol, and drug reaction. DATA SYNTHESIS: The manufacturer's warnings include a disulfiram-like reaction between metronidazole and ethanol. However, review of reports published between 1969 and 1982 produced no convincing evidence that this reaction exists. Six case reports involving eight patients were evaluated. CONCLUSIONS: Four of the eight cases were serious, including one death, but the authors of all the reports presumed the metronidazole-ethanol reaction to be an established pharmacologic fact. None provided evidence that could justify their conclusions. PMID- 10676836 TI - Lamotrigine for the treatment of bipolar disorder. AB - OBJECTIVE: To describe the available data regarding the clinical efficacy of lamotrigine for the treatment of bipolar disorder. SUMMARY: Anticonvulsants have emerged as alternative mood-stabilizing agents for patients with bipolar disorder who do not respond to lithium. Data regarding the efficacy of lamotrigine have been generated primarily from case reports, small open trials, and one large, randomized, placebo-controlled trial. These reports suggest that lamotrigine may be effective for the management of bipolar disorder. CONCLUSIONS: Although current data are limited, treatment-refractory patients with bipolar disorder may benefit from lamotrigine therapy. Several studies are currently underway to determine the appropriate role of lamotrigine in the treatment of bipolar disorder. PMID- 10676837 TI - Fever, rash, and pancytopenia following vancomycin rechallenge in the presence of ceftazidime. PMID- 10676838 TI - Osteomalacia associated with carbamazepine/valproate. PMID- 10676839 TI - Psychotropic medication use in older patients referred for evaluation of falls risk. PMID- 10676840 TI - de Clerambault syndrome successfully treated with olanzapine. PMID- 10676841 TI - Withdrawal-emergent dyskinesia in a patient taking risperidone/citalopram. PMID- 10676842 TI - The p38 signal transduction pathway: activation and function. AB - The p38 signalling transduction pathway, a Mitogen-activated protein (MAP) kinase pathway, plays an essential role in regulating many cellular processes including inflammation, cell differentiation, cell growth and death. Activation of p38 often through extracellular stimuli such as bacterial pathogens and cytokines, mediates signal transduction into the nucleus to turn on the responsive genes. p38 also transduces signals to other cellular components to execute different cellular responses. In this review, we summarize the characteristics of the major components of the p38 signalling transduction pathway and highlight the targets of this pathway and the physiological function of the p38 activation. PMID- 10676843 TI - Induction of protein kinase Czeta-related protein kinase by growth suppression in carcinogen-initiated epidermal cell-line WYF31 cells. AB - In primary cultured mouse epidermal cells, protein kinase C isozyme zeta (PKCzeta) consists of multiple forms, for example, low-salt eluted PKCzeta (1 PKCzeta; 79 and 85 kDa) and high-salt eluted PKCzeta (h-PKCzeta; 79 and 85 kDa) on anion-exchange column chromatography. In this study, biochemical and biophysical differences between 1-PKCzeta and h-PKCzeta were examined by using carcinogen-initiated mouse epidermal cell-line WYF31 cells, whose growth is stimulated by tumour promoter phorbol 12-myristate 13-acetate (PMA). The binding efficiency of h-PKCzeta to anti-PKCzeta antibody-affinity column was 10 times higher than that of 1-PKCzeta. T7-tagged rat PKCzeta overexpressed in WYF31 cells was recovered only in the high-salt eluted area on the anion-exchange column. Furthermore, when rat PKCzeta was stably overexpressed in WYF31 cells, the content of h-PKCzeta increased 4 to 5 times compared to that of parental cells, but the content of 1-PKCzeta was not altered. All of these results indicate that h-PKCzeta is the product of the PKCzeta gene (referred to as PKCzeta) and that 1 PKCzeta is closely related but different from PKCzeta (referred to as PKCzeta related kinase). Interestingly, serum starvation of WYF31 cells caused a marked increase of the content of PKCzeta-related kinase with a concomitant decrease of PKCzeta content. These changes were reversed by stimulating the cell growth with 10% foetal calf serum. Prolonged treatment of starved cells with PMA, which induces the proliferation of WYF31 cells, also caused the downregulation of PKCzeta-related kinase. These results suggest that the expression levels of PKCzeta-related kinase and PKCzeta are differently regulated, and that the increased expression of PKCzeta-related kinase might play a significant role in the growth-suppression processes of WYF31 cells. PMID- 10676844 TI - Activation of STAT5 during EPO-directed suppression of apoptosis. AB - The ligand-dependent activation of the JAK/STAT (Januskinase/Signal Transducer and Activator of Transcription) pathway has been implicated in the explanation of cytokine-specific regulation of gene expression. Previous studies have reported conflicting results on the role of the transcription factor STAT5 in erythropoietin (EPO)-induced cellular responses. In this study we focused on the functional importance of STAT5 docking sites in the intracellular EPO receptor (EPOR) domain for the mediation of antiapoptotic activities. We demonstrate that EPO-dependent survival of erythroleukemic cell lines is accompanied by sustained STAT5 DNA-binding activity. The role of single tyrosine residues was dissected by the analysis of myeloid FDCP-1 cells stably expressing mutant EPOR proteins. The data show that receptors having a high potential to mediate antiapoptotic signals also effectively activate STAT5, whereas receptors lacking STAT5 docking sites are diminished in both activities. We conclude that the transcription factor STAT5 is functionally implicated in the EPO-dependent survival of cells. PMID- 10676845 TI - Caffeine exerts a dual effect on capacitative calcium entry in Xenopus oocytes. AB - Caffeine increases the amplitude of the Cl- currents evoked by capacitative Ca2+ entry (CCE) on thapsigargin-treated Xenopus oocytes. The caffeine-induced potentiation of the CCE process appears to rest on two distinct and additive components. The first component involves the cAMP second messenger system since it can be mimicked by either IBMX perfusion or cAMP microinjection into the oocyte and inhibited by the PKA inhibitory peptide i-PKA. The second component, although activatory, is dynamically related to the caffeine-evoked inhibition of InsP3-mediated Ca+ release and may arise from an interaction between caffeine and the InsP3 receptor in the context of a conformational coupling between the InsP, receptor and the channels responsible for CCE. PMID- 10676846 TI - Altered activation of phospholipase D by lysophosphatidic acid and endothelin-1 in mouse embryo fibroblasts lacking phospholipase C-gamma1. AB - Lysophosphatidic acid (LPA) and endothelin-1 (ET-1) activate phospholipase D (PLD) in many cell types. To see if phospholipase C-gamma1 plays a role, we used embryonic fibroblasts from mice in which the PLCgamma1 gene was disrupted. Surprisingly, the effect of LPA on inositol phosphate accumulation was increased in these PLCgamma1-/- cells, whereas that of ET-1 was completely abrogated. When PLD activity was measured, the response to LPA was also enhanced and the response to ET-1 lost in the PLCgamma1-/- cells. Treatment of these cells with ionomycin and oleoyl acetyl glycerol to mimic PLC stimulation restored PLD activity. Treatment of either PLCgamma1+/+ and PLCgamma1-/- cells with tyrosine kinase inhibitors did not inhibit LPA- or ET-1-induced PLD activity. Moreover, LPA and ET-1 treatment of PLCgamma1+/+ and PLCgamma1-/- cells did not cause tyrosine phosphorylation of PLC-gamma1 or PLC-gamma2. In summary, these results show that the altered PLD responses to LPA and ET-1 in PLCgamma1-/- are due to changes in PLC activity and do not involve tyrosine kinase activity. PMID- 10676847 TI - Acute modulation of Ca2+ influx on rat heart by 17beta-estradiol. AB - Estrogens initiate their action by binding to specific intracellular receptors and then acting on gene expression. In addition, there is growing evidence of a direct membrane effect via interaction with a cell surphase receptor. The aim of the present study was to investigate the acute effects of 17beta-estradiol on Ca2+ fluxes through second messenger pathways in rat cardiac muscle. Exposure of rat ventricle to low levels of 17beta-estradiol (10(-12)-10(-8) M) increased 45Ca2+ influx within 1 min (+38%); the response was biphasic, peaking at 2 and 5 min (+60 and +55%, respectively). The effect of the hormone on rat heart seems to be specific since 17alpha-estradiol, dihydrotestosterone, and progesterone were devoid of activity. The effect of 17beta-estradiol (5 min, 10(-10) M) was suppressed by nitrendipine (1 microM) and LaCl3 (10 microM), involving the activation of voltage-dependent Ca2+ channels in the acute increase of rat heart calcium influx by the hormone. 17Beta-estradiol rapidly increased cAMP content and PKA activity of rat cardiac muscle in parallel to the changes in Ca2+ uptake. In addition the cAMP antagonist Rp-cAMPS suppressed 17beta-estradiol-dependent Ca2+ influx. Altogether, the data suggest the involvement of the cAMP/PKA messenger system in the nongenomic modulation of Ca2+ influx in rat cardiac muscle by physiological levels of 17beta-estradiol. PMID- 10676848 TI - Divergent proliferative responses to a gastrin receptor ligand in synchronized and unsynchronized rat pancreatic AR42J tumour cells. AB - Depending upon experimental model, the CCK-B/gastrin receptor ligand CI-988 exhibits either agonist or antagonist activity. To confirm that CI-988 behaves as an antagonist toward gastrin-stimulated growth, its effects on cell proliferation were investigated in unsynchronized and synchronized AR42J rat pancreatic tumour cells. In unsynchronized cultures CI-988 alone had no effect, but inhibited gastrin-stimulated cell proliferation. In contrast, in synchronized cultures, CI 988 stimulated cell proliferation. Similarly, CI-988 inhibited gastrin-stimulated cAMP production in unsynchronized cells, but stimulated cAMP formation in synchronized cultures. Therefore, CI-988 stimulation of cAMP production and proliferation in AR42J cell cultures appears to be cell cycle-dependent. CI-988 inhibited gastrin-stimulated intracellular calcium ([Ca2+]i) mobilization in both populations and thus acted as an antagonist toward this pathway. Because CCK receptor densities and affinities were similar in both cell populations, the data suggest that CI-988's divergent effects on cell proliferation are governed by postreceptor signalling events which vary with cell cycle. PMID- 10676849 TI - Involvement of phosphorylation of beta-subunit in cAMP-dependent activation of L type Ca2+ channel in aortic smooth muscle-derived A7r5 cells. AB - We investigated the effect of intracellular cAMP on the gating kinetics of L-type Ca2+ channel in an A7r5 smooth muscle-derived cell line using the whole-cell patch-clamp technique. Application of dibutyryl cyclic AMP (db-cAMP) to the cell increased the magnitude of Ca2+ currents through L-type Ca2+ channels (I(Ca)), and shifted the current-voltage relationship (I-V curve) for I(Ca) to the left. The magnitudes of maximum I(Ca) were 14.1 +/- 0.7 before and 16.0 +/- 1.1 pA/pF after application of 1 mM db-cAMP (P < 0.05). The values of the half-activation potential (V(1/2)) of I(Ca), estimated from activation curves, were -7.0 +/- 0.8 mV before and -10.8 +/- 1.0 mV after application of db-cAMP (P < 0.05). In cells pretreated with 10 microM Rp-cAMPS (a specific inhibitor of PKA), db-cAMP affected neither the I-V curve nor the activation curve for I(Ca). In cells pretreated with the antisense oligonucleotide for the beta-subunit of L-type Ca2+ channel, db-cAMP failed to enhance I(Ca) or alter the activation curve. On the other hand, in the cells pretreated with the nonsense oligonucleotide, application of db-cAMP caused an increase in magnitude of I(Ca) and shifted the activation curve to the left. Western blot analysis revealed that the pretreatment of cells with antisense oligonucleotide but nonsense oligonucleotide reduced the expression of the beta-subunit of the L-type Ca2+ channel. We conclude that the cAMP-dependent phosphorylation of the beta-subunit potentiates the voltage dependency of the activation kinetics of the L-type Ca2+ channel in A7r5 cells. PMID- 10676850 TI - What the evolution of the amyloid protein precursor supergene family tells us about its function. AB - The Alzheimer's disease amyloid protein precursor (APP) gene is part of a multi gene super-family from which sixteen homologous amyloid precursor-like proteins (APLP) and APP species homologues have been isolated and characterised. Comparison of exon structure (including the uncharacterised APL-1 gene), construction of phylogenetic trees, and analysis of the protein sequence alignment of known homologues of the APP super-family were performed to reconstruct the evolution of the family and to assess the functional significance of conserved protein sequences between homologues. This analysis supports an adhesion function for all members of the APP super family, with specificity determined by those sequences which are not conserved between APLP lineages, and provides evidence for an increasingly complex APP superfamily during evolution. The analysis also suggests that Drosophila APPL and Caenorhabditis elegans APL-1 may be a fourth APLP lineage indicating that these proteins, while not functional homologues of human APP, are similarly likely to regulate cell adhesion. Furthermore, the betaA4 sequence is highly conserved only in APP orthologues, strongly suggesting this sequence is of significant functional importance in this lineage. PMID- 10676851 TI - In-vivo glutathione elevation protects against hydroxyl free radical-induced protein oxidation in rat brain. AB - Glutathione deficiency has been associated with a number of neurodegenerative diseases including Lou Gehrig's disease, Parkinson's disease, and HIV. A crucial role for glutathione is as a free radical scavenger. Alzheimer's disease (AD) brain is characterized by oxidative stress, manifested by protein oxidation, lipid oxidation, oxidized glutathione, and decreased activity of glutathione S transferase, among others. Reasoning that elevated levels of endogenous glutathione would offer protection against free radical-induced oxidative stress, rodents were given in vivo injections of N-acetylcysteine (NAC), a known precursor of glutathione, to study the vulnerability of isolated synaptosomal membranes treated with Fe2+/H2O2, a known hydroxyl free radical producer. Protein carbonyls, a marker of protein oxidation, were measured. NAC significantly increased endogenous glutathione levels in cortical synaptosome cytosol (P < 0.01). As reported previously, protein carbonyl levels of the Fe2+/H2O2-treated synaptosomes were significantly higher compared to that of non-treated controls (P < 0.01), consistent with increased oxidative stress. In contrast, protein carbonyl levels in Fe2+/H2O2-treated synaptosomes isolated from NAC-injected animals were not significantly different from saline-injected non-treated controls, demonstrating protection against hydroxyl radical induced oxidative stress. These results are consistent with the notion that methods to increase endogenous glutathione levels in neurodegenerative diseases associated with oxidative stress, including AD, may be promising. PMID- 10676852 TI - Analgesic effect of interferon-alpha via mu opioid receptor in the rat. AB - Using the tail-flick induced by electro-stimulation as a pain marker, it was found that pain threshold (PT) was significantly increased after injecting interferon-alpha (IFN alpha) into the lateral ventricle of rats. This effect was dosage-dependent and abolished by monoclonal antibody (McAb) to IFN alpha. Naloxone could inhibit the analgesic effect of IFN alpha, suggesting that the analgesic effect of IFN alpha be related to the opioid receptors. Beta funaltrexamine (beta-FNA), the mu specific receptor antagonist could completely block the analgesic effect of IFN alpha. The selective delta-opioid receptor antagonist, ICI174,864 and the kappa-opioid receptor antagonist, nor-BNI both failed to prevent the analgesic effect of IFN alpha. IFN alpha could significantly inhibit the production of the cAMP stimulated by forskolin in SK-N SH cells expressing the mu-opioid receptor, not in NG108-15 cells expressing the delta-opioid receptor uniformly. The results obtained provide further evidence for opioid activity of IFN alpha and suggest that this effect is mediated by central opioid receptors of the mu subtype. The evidence is consistent with the hypothesis that multiple actions of cytokines, such as immunoregulatory and neuroregulatory effects, might be mediated by distinct domains of cytokines interacting with different receptors. PMID- 10676853 TI - Serotonin transporter function is modulated by brain-derived neurotrophic factor (BDNF) but not nerve growth factor (NGF). AB - The serotonin transporter (5-HTT) regulates serotonergic neurotransmission by determining the magnitude and duration of serotonergic responses. We have recently described a polymorphism in the 5-HTT gene promoter (5-HTTLPR) which influences the function of the 5-HTT and is associated with several psychiatric disorders. Immortalized B lymphocytes express the 5-HTT, and a B lymphocyte line has been shown to express the receptor for brain-derived neurotrophic factor, trkB. Since brain-derived neurotrophic factor (BDNF) is a specific growth and differentiation factor for serotonergic neurons, we assessed whether BDNF is able to modulate 5-HTT function in B lymphoblasts. Nerve growth factor (NGF), another neurotrophin which acts via the trkA receptor, was also studied. Eight immortalized B lymphoblast lines were generated and genotyped for the 5-HTTLPR. After treatment with BDNF or NGF, 5-HT uptake and proliferation of the cell lines were assessed. Two of the B cell lines showed a dose-dependent reduction of 5-HT uptake after exposure to BDNF. Both of these cell lines were homozygous for the long allele of the 5-HTTLPR. NGF did not influence 5-HT uptake or cellular proliferation in any of the cell lines. Thus, BDNF but not NGF may influence 5-HT uptake in some B lymphocytes. The fact that regulation of the 5-HTT was observed preferentially in cells of the long/long genotype indicates that presence of a short allele confers reduced regulatory capacity on the 5-HTT. In conclusion, B lymphoblasts represent a practical model for functional regulation of the 5-HTT by neurotrophins in serotonergic neurons. PMID- 10676854 TI - Stimulation of Na+,K+-ATPase activity, increase in potassium uptake, and enhanced production of ouabain-like compounds in ammonia-treated mouse astrocytes. AB - Active potassium (K+) uptake and Na+,K+-ATPase activity were measured in primary cultures of mouse astrocytes. Both parameters were virtually unaffected by acute ammonia treatment but increased after chronic exposure to pathophysiologically relevant concentrations of ammonia (0.3 or 3 mM) for 1-4 days. The increased Na+,K+-ATPase activity after chronic treatment with ammonia was further enhanced in the acute presence of 12 mM K+. Based on these observations and literature data it was hypothesized that the direct effect of ammonia is formation of easily diffusible compound(s) with ouabain-like effect, that upregulation occurs of Na+,K+-ATPase activity and K+ uptake in response to the resulting ATPase inhibition, and that the washing procedure preceding the uptake experiments and the determination of Na+,K+-ATPase activity unmasks the upregulation. To test this hypothesis, the content of compounds with ouabain-like action was measured in media in which astrocytes had been incubated in the presence of 3 mM ammonia for 4 days and in controls to which an additional 3 mM NaCl had been added instead of ammonia. An endogenous, compound with ouabain-like activity was demonstrated both under control conditions and in the ammonia-treated cultures, and the content of this compound was increased by 50% in the ammonia-treated cultures. Preliminary experiments showed that at least part of the released ouabain-like compounds cross-react with authentic ouabain. PMID- 10676855 TI - Melatonin inhibits pituitary adenylyl cyclase-activating polypeptide-induced increase of cyclic AMP accumulation and [Ca2+]i in cultured cells of neonatal rat pituitary. AB - The effects of melatonin on pituitary adenylyl cyclase-activating polypeptide induced increase of cyclic AMP and [Ca2+]i were studied in neonatal rat pituitary cells. The polypeptide increased cyclic AMP accumulation. In the presence of melatonin the increase of cyclic AMP was inhibited in a dose-dependent manner, the maximal inhibition was achieved with 1-10 nM melatonin. Pituitary adenylyl cyclase-activating polypeptide also increased [Ca2+]i in 30% of the pituitary cells and melatonin inhibited the effect. Most of the cells sensitive to adenylyl cyclase-activating polypeptide (77%) were also sensitive to GnRH, suggesting they are gonadotrophs. The remaining cells were not identified. The polypeptide induced [Ca2+]i increase was inhibited in Ca2+-free medium in 2/3 of the cells indicating that Ca2+ influx was involved. To examine causal relationship between cyclic AMP and [Ca2+]i increase, we have studied the effect of adenylyl cyclase activation by forskolin on intracellular Ca2+ concentration. Forskolin had similar effects as adenylyl cyclase-activating polypeptide: it increased [Ca2+]i in the pituitary cells and the increase was dependent on presence of Ca2+ in the medium. Melatonin inhibited the forskolin induced [Ca2+]i increase. Our observations indicate that increase of cyclic AMP stimulates Ca2+ influx in the pituitary cells of neonatal rat and that this mechanism is involved in [Ca2+]i increase induced by the pituitary adenylyl cyclase-activating polypeptide. Because melatonin inhibits increase of cyclic AMP induced by pituitary adenylyl cyclase-activating polypeptide or forskolin, the inhibitory effect of melatonin on Ca2+-influx may be mediated by the decrease of cyclic AMP concentration. This mechanism of melatonin action has not been described previously. Because melatonin inhibits the polypeptide- or forskolin-induced [Ca2+]i also in the cells not sensitive to GnRH, melatonin receptors seem to be present on both gonadotrophs and non-gonadotrophic pituitary cells. PMID- 10676856 TI - Difference in glutamate release between retina and cerebral cortex following ischemia. AB - The difference in ischemic tolerance between the retina and cerebral cortex may be attributable to a difference in glutamate release during ischemia. Glutamate release in the retina and the cerebral cortex was compared in rats. A dialysis electrode for real-time glutamate measurement was perfused with L-glutamate oxidase, and the current evoked between two voltage-clamped electrodes was detected. Two electrodes were implanted in the retina through the choroid and cerebral cortex in 12 anesthetized rats, each mounted on a stereotaxic frame. Global ischemia was induced by ligation on both carotid arteries and hypotension was induced by blood withdrawal. Under control conditions, the glutamate concentration in the retina was 164 +/- 231 (mean +/- standard deviation) microM, being significantly higher (P < 0.05) than that in the cerebral cortex (83 +/- 105 microM). In 10 of the 12 animals, the glutamate concentration in the retina decreased to a minimum of 134 +/- 149 microM (P < 0.01, compared with the value for the cerebral cortex), but that in the cortex increased to 410 +/- 305 microM (averaged highest value). Immediately after the start of reperfusion, the glutamate concentration in the cortex decreased rapidly to 101 +/- 27 microM, but that in the retina increased gradually to almost the control level (148 +/- 204 microM). In the other two animals, the glutamate concentration remained unchanged. In conclusion, glutamate release in the retina does not proceed as rapidly as that in the cerebral cortex during 20 min of ischemia, and in fact decreases. This opposite trend shown by the two organs may be due to the slow depletion rate of ATP in the retina. This may explain the differing neuronal tolerance to ischemia in these two organs. PMID- 10676857 TI - Lack of stereoselectivity of 8-hydroxy-2(di-N-propylamino)tetralin-mediated inhibition of forskolin-stimulated adenylyl cyclase activity in human pre- and post-synaptic brain regions. AB - The stereoselectivity of the serotonin1A (5-HT1A) receptor compound 8-hydroxy 2(di-N-propylamino)tetralin (8-OH-DPAT) on forskolin-stimulated adenylyl cyclase activity was investigated in membranes from human 5-HT pre-synaptic (raphe nuclei) and post-synaptic (hippocampus and prefrontal cortex) regions of autopsy brains. After sample incubation with agonists and antagonists, results showed that both the racemic mixture of 8-OH-DPAT or its (+) and (-) enantiomers behaved as full agonists in the tested brain regions. Enantiomer potency (EC50, nM) and efficacy (percentage of maximal inhibition, %) values were similar in all regions under investigation. However, some inter and intra-region variations in racemic 8 OH-DPAT potency and efficacy have been observed. In particular, the potency of racemic 8-OH-DPAT was higher in the prefrontal cortex and raphe nuclei than in the hippocampus, where it was in fact lower than either single enantiomers. Agonist effects were competitively reversed by 5-HT1A antagonists, although once again a different profile was revealed in the hippocampus. The data underscores the lack of stereospecificity of 8-OH-DPAT-mediated inhibition of adenylyl cyclase activity in either pre- or post-synaptic human brain regions. Moreover, such results have significant implication, as they support the notion that human 5-HT1A receptors might vary from one brain region to the other. PMID- 10676858 TI - Mitochondrial impairment and recovery after heat shock treatment in a human microglial cell line. AB - The application of a heat shock on the human microglial cell line (CHME 5) has been shown to cause cytoskeleton modifications and alterations in phosphorylated metabolite content (Macouillard-Poulletier de Gannes et al., 1998a Metabolic and cellular characterization of immortalized human microglial cells under heat stress. Neurochem. Int. 33, 61-73). In this study, we focused on the possible involvement of mitochondria in this heat stress response. The cell respiratory properties were followed during the recovering period and the possible relationships between mitochondria and the cytoskeleton were studied. We observed that the heat shock induced changes in mitochondrial activity due to protein denaturation, rather than mitochondrial loss. Furthermore, these alterations were correlated with cytoskeleton disorganization since vimentine, tubuline and mitochondria shift, simultaneously, to a perinuclear location. The perturbations of the mitochondrial distribution persisted until cytoskeleton networks had recovered. Nevertheless, the respiratory properties recovered rapidly suggesting a renaturation of mitochondrial proteins in connection with mitochondrial cytoplasmic redistribution. PMID- 10676859 TI - Effects of an adenosine analogue administration on the striatal NMDA receptors in an experimental model of epilepsy. AB - Specific [3H]-MK801 binding to rat NMDA receptors following the administration of the convulsant drug 3-mercaptopropionic acid (MP) and the adenosine analogue cyclopentyladenosine (CPA) was studied in striatal membrane fractions. MP administration (150 mg/kg, i.p.) caused an increase of 53% and 82% in [3H]-MK801 binding during seizure and the postseizure period respectively. Administration of CPA (2 mg/kg, i.p.) raised [3H]-MK801 binding by 72%. When CPA was administered 30 min before MP and rats sacrificed at seizure (CPA + MPc), an increase of 64%, was observed. Saturation results indicate that receptor sites increased their maximal binding capacity (Bmax) in all treatments while the apparent dissociation constant (Kd) remained unchanged. MP administration brought about an increase of 52% and 42% in [3H]-MK801 binding sites during seizure and postseizure respectively. Administration of CPA raised receptor density by 75%. When CPA was administered 30 min before MP and rats sacrificed at seizure (CPA + MPc), an increase of 62%, was observed. These results show that striatal NMDA receptors have a selective role in seizure activity in the basal ganglia and that the adenosine analogue administration may modify [3H]-MK801 binding in a way similar to that of the convulsant drug. PMID- 10676860 TI - Uncompetitive stimulation of rat brain Na-K ATPase activity by rapid eye movement sleep deprivation. AB - Rapid eye movement sleep deprivation is associated with an increase in Na-K ATPase activity. In order to understand the possible biochemical mechanism of this increase, the kinetics of Na-K ATPase was studied. Although the enzyme activity increased after the deprivation, the catalytic efficiency of the enzyme remained unaltered. The rapid eye movement sleep deprivation increased both the Vmax and the Km suggesting an uncompetitive stimulation of the enzyme. While increase in norepinephrine resulted in an increased Vmax, that of calcium increased the Km. Since an increase in norepinephrine has been suggested after deprivation, the increased Vmax is attributed to increased norepinephrine level following deprivation. However, since rapid eye movement sleep deprivation is reported to be associated with a decrease in calcium levels, the increase in Km following deprivation may be attributed to changes in factor(s) other than calcium. PMID- 10676861 TI - Adenosine modulation of D-[3H]aspartate release in cultured retina cells exposed to oxidative stress. AB - In this study we evaluated the role of adenosine receptor activation on the K+ evoked D-[3H]aspartate release in cultured chick retina cells exposed to oxidant conditions. Oxidative stress, induced by ascorbate (3.5 mM)/Fe2+ (100 microM), increased by about fourfold the release of D-[3H]aspartate, evoked by KCl 35 mM in the presence and in the absence of Ca2+. The agonist of A1 adenosine receptors, N6-cyclopentyladenosine (CPA; 10 nM), inhibited the K+-evoked D [3H]aspartate release in control in oxidized cells. The antagonist of A1 adenosine receptor, 1,3-dipropyl-8-cyclopentylxanthine (DPCPX; 50 nM), potentiated the release of D-[3H]aspartate in oxidized cells, and reverted the effect observed in the presence of CPA 10 nM. However, in oxidized cells, when DPCPX was tested together with CPA 100 nM the total release of D-[3H]aspartate increased from 5.1 +/- 0.4% to 11.4 +/- 1.0%, this increase being reverted by 3,7 dimethyl-1-propargylxanthine (DMPX; 100 nM), an antagonist of A2A adenosine receptors. In cells of both experimental conditions, the K+-evoked release of D [3H]aspartate was potentiated by the selective agonist of A2A adenosine receptors, 2-[4-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamidoadenosin e (CGS 21680; 10 nM), whereas the antagonist of these receptors, DMPX (100 nM), inhibited the release of D-[3H]aspartate in oxidized cells, but not in control cells. Adenosine deaminase (ADA; 1 U/ml), which is able to remove adenosine from the synaptic space, reduced the K+-evoked D-[3H]aspartate release, from 5.1 +/- 0.4% to 3.1 +/- 0.3% in oxidized cells, and had no significant effect in control cells. The extracellular accumulation of endogenous adenosine, upon K+ depolarization, was higher in oxidized cells than in control cells, and was reduced by the inhibitors of adenosine transporter (NBTI) and of ecto-5' nucleotidase (AOPCP). This suggests that adenosine accumulation resulted from the outflow of adenosine mediated by the transporter, and from extracellular degradation of adenine nucleotide. Our data show that both inhibitory A1 and excitatory A2A adenosine receptors are present in cultured retina cells, and that the K+-evoked D-[3H]aspartate release is modulated by the balance between inhibitory and excitatory responses. Under oxidative stress conditions, the extracellular accumulation of endogenous adenosine seems to reach levels enough to potentiate the release of D-[3H]aspartate by the tonic activation of A2A adenosine receptors. PMID- 10676862 TI - Impact of modifying priming components and fluid administration using miniaturized circuitry in neonatal cardiopulmonary bypass. AB - Following a succession of changes in circuitry and priming additives between 1993 and 1998, a comprehensive re-evaluation of neonatal cardiopulmonary bypass (CPB) practice was undertaken. Samples from 10 infants (Group 1) undergoing CPB were evaluated for osmolality, oncotic pressure, total protein, hematocrit, glucose, and electrolytes (Na+, K+, iCa2+). These samples were tested at six measurement points: (1) after priming, (2) patient pre-CPB, (3) CPB-start, (4) CPB-mid, (5) CPB-end, and (6) post-modified ultrafiltration (MUF). Prime volumes were also carefully measured as well as the type and amount of volume given during CPB. After evaluating the initial data, changes in protocol regarding mannitol, calcium correction, and oncotic strength on CPB were made. Following implementation of these protocol changes, a second set (Group 2) of 10 infants was identically evaluated. Group 1 prime osmolality was 379 +/- 44 mOsm/kg, while Group 2 prime osmolality was 324 +/- 14 mOsm/kg (p = 0.003). There were no differences in osmolality between groups during bypass and osmolality was unaffected by modified ultrafiltration. Ionized calcium levels were significantly different at the end of bypass between Group 1, 0.6 +/- 0.1 mmol/l; and Group 2, 1.17 +/- 0.24 mmol/l (p < 0.001). In Group 1, there was a 40% drop (p = 0.001) in colloid osmotic pressure (COP) levels from pre-CPB (13.3 +/- 3.4 mmHg) to CPB-end (8.8 +/- 1.2 mmHg). In Group 2, there were no differences in COP during CPB. COP levels of Group 1 and Group 2 at CPB-end were 8.8 +/- 1.2 mmHg and 14 +/- 1.9, respectively (p < 0.0001). Total volume addition during bypass for Group 1 was 363.5 +/- 148.7 ml and for Group 2 was 245.1 +/- 92.2 ml (p < 0.05). In conclusion, progressive changes in neonatal circuits and techniques can have potentially wide-ranging effects on electrolyte and osmotic/oncotic homeostasis. An audit of perfusion management through expanded laboratory tests is recommended, especially in periods of change. PMID- 10676863 TI - Comparison of Sarns 3M heparin bonded to Duraflo II and control circuits in a porcine model: macro- and microanalysis of thrombi accumulation in circuit arterial filters. AB - Heparin-bonded perfusion circuits have been reported to reduce the thrombus formation during various levels of systemic heparinization. The goal of this study was to compare the efficacy of thrombo-resistance of the Sarns 3M heparin bonded circuit to Baxter Duraflo II and untreated control in a porcine model. Fifteen Yorkshire pigs (60-65 kg) were anesthetized, heparinized with 3000 IU, intravenously (i.v.) and surgically cannulated with an internal jugular outflow and a femoral vein inflow. All circuits consisted of a 22-Fr venous cannula, centrifugal pump, arterial filter, an 18-Fr cannula for return and connected with equal lengths of 3/8" polyvinyl chloride tubing. The flows were maintained at 2.0 l/min for 4 h. Thrombus formation in filter samples were morphometrically analyzed through macro-densitometry, light microscopy, and scanning electron microscopy (SEM). Our findings revealed that the 3M circuit had significantly less gross thrombus (p < 0.001), 66% and 84% less microscopic thrombi and fivefold less SEM-measured aggregates (p = 0.03) compared to the Duraflo II and uncoated groups. This study demonstrated that the 3M heparin-bonded circuit had significantly reduced the formation of micro- and macro-thrombi in the minimally heparinized pig model compared to the Duraflo II and untreated control circuits. PMID- 10676864 TI - In vitro evaluation of sedative drug losses during extracorporeal membrane oxygenation. AB - Sedative agents are routinely administered to critically ill patients, both on and off extracorporeal membrane oxygenation (ECMO), to enable patients to be comfortable and facilitate patient management. It has been observed empirically in our paediatric intensive care unit that doses of sedative drugs required to achieve desired levels of sedation in ECMO patients are far greater than those used in non-ECMO patients. These differences could not simply be accounted for by differences in patient types, clinical status or sedation levels. We therefore undertook an in vitro evaluation of drug binding in ECMO circuits. This study investigated how the polyvinyl chloride (PVC) and silicone rubber components of neonatal ECMO circuits affect drug delivery in patients through drug sorption. Phase 1 investigated drug uptake by the two polymers in static solutions of known concentrations of four commonly used sedative drugs: lorazepam, midazolam, diazepam and propofol. Phase 2 involved the setting up of a complete neonatal ECMO circuit, injecting the drug solutions pre reservoir at a flow rate of 350 ml/min and collecting samples post-oxygenator for analysis. Phase 1 results revealed significant uptake of drugs with losses in the range 40-98% and in the order propofol > diazepam > midazolam > orazepam. Phase 2 results were similar and in the first 40 min of running an ECMO circuit only 10% of propofol passed through the circuit. These results may help to explain observed clinical phenomena and raise important issues regarding drug dosing in ECMO patients. PMID- 10676865 TI - The hazardous effects of alveolar hypocapnia on lung mechanics during weaning from cardiopulmonary bypass. AB - The bronchoconstrictive effects of alveolar hypocapnia during weaning from cardiopulmonary bypass (CPB) were investigated in patients undergoing elective coronary artery revascularization. Thirty patients were randomly assigned into two equal groups. In both groups, mechanical ventilation was initiated for 3 min prior to weaning from CPB with the venous pressure low. This kept the pulmonary vascular bed empty, resulting in alveolar hypocapnia (ETCO2 < 2 kPa). Peak airway pressure (P(peak)) and plateau pressures (P(plateau)) were recorded. In group 1, 5% CO2 was added to the inspiratory gas mixture and the ETCO2 allowed to rise (ETCO2 > 3.3 kPa). The ventilation pressure measurements were recorded again after 3 min stabilization. In group 2, the venous pressure was increased to allow the pulmonary venous bed to fill and the ventilation pressures recorded after a 3 min period of stabilization. In group 1, the ventilatory pressures dropped significantly (p < 0.001) when the alveolar hypocapnia was reversed with added CO2 (P(peak) 19.71 +/- 5.7 to 12.31 +/- 2.8 cmH2O and P(plateau) 13.15 +/- 3.28 to 9.15 +/- 2.23 cmH2O). In group 2, a similar effect was achieved by allowing filling of the pulmonary vascular bed (P(peak) 17.46 +/- 4.72 to 11.92 +/- 3.03 cmH2O and P(plateau) 13.93 +/- 4.10 to 9.37 +/- 3.00 cmH2O). These results suggest that filling the pulmonary vascular bed prior to initiating ventilation, when weaning from CPB, prevents the otherwise deleterious effects of alveolar hypocapnia, resulting in raised bronchomotor tonus and raised airway pressures. PMID- 10676866 TI - Red blood cell trauma during cardiopulmonary bypass: narrow pore filterability versus free haemoglobin. AB - Ten patients admitted for coronary artery bypass grafting were investigated with respect to the influence of cardiopulmonary bypass (CPB) on red blood cell (RBC) trauma. Blood samples were collected prior to, at the start of, and at 30 and 60 min of CPB. RBC deformability was assessed by filtering re-suspended RBCs through a polycarbonate membrane using a computer-controlled filtrometer. Multiple regression analysis was employed to evaluate RBC flow-curve characteristics denoted by the initial filtration rate (IFR) and clogging slope (CS). Release of free haemoglobin was determined concomitantly. IFR was estimated at 90.39 microl/s and CS at -5.32 microl/s2 prior to CPB. During 60 min of CPB, neither IFR nor CS deviated significantly (p > 0.05) from these reference values. However, release of free haemoglobin increased significantly (p < 0.018) from the start of CPB to the 60-min determination. In conclusion, 60 min of CPB seems not to alter significantly RBC deformability in a 5 microm pore filtration model, despite a significant release of haemoglobin. PMID- 10676867 TI - Platelets and whole blood coagulation. AB - In our early work in developing activated clotting time (ACT) assays, it became apparent that changes occurred in coagulation times as a whole blood sample aged (0-6 h). Subsequent studies showed that the coagulation parameters of plasma obtained from the samples remained stable during this time frame. These changes in whole blood clotting times during storage were eventually traced to the platelets. Several years of work demonstrated that this change was due to the removal of the blood from the vascular lining. This recalled a mechanism that was originally put forth in the 1970s with the discovery of prostacyclin. In this postulated mechanism, platelets are 'time-bombs'. They are kept under control by prostacyclin (PGI2) secreted by the vascular lining. Without this prostacyclin, platelets 'preactivate'. Since that time, additional substances secreted by the vascular endothelium have been identified, such as nitric oxide, that also influence platelet activity. The 'preactivation' of platelets in a blood sample can be followed using an ACT. In the same donor, the preactivation is uniform and reproducible over an extended period (months). There is, however, considerable variability between donors. Some donors' platelets preactivate dramatically, while other donors show hardly any change. Prostacyclin, added to the blood sample when it is collected, prevents this preactivation. The clinical significance of these observations has yet to be clearly established, but these observations raise a number of questions with respect to methods for improving platelet function during bypass and in evaluating the risk of platelet-mediated cardiovascular disease. PMID- 10676868 TI - A retrospective study on perfusion incidents and safety devices. AB - Despite the acceptance of extracorporeal circulation as an effective modality to facilitate cardiac surgery, patient outcomes can be negatively influenced by the occurrence of perfusion incidents. A perfusion survey was conducted to identify safety techniques and incidents related to cardiopulmonary bypass (CPB). An 80 question survey was mailed to chief perfusionists of all 1030 USA cardiac surgical centers using CPB. The survey was designed to examine practices and incidents that occurred during a 2-year period (July 1996 to July 1998). Five hundred-and-fifty-two (54% response rate) surveys were returned, which accounted for 797 hospitals (79% of all cardiac centers) and 653,621 surgical procedures. Of the 27 identified CPB safety devices, the highest utilization was arterial line filters (98.5%) and the lowest arterial line bubble traps (3.4%). Of the reported cases, a CPB incident occurred once every 138 cases. The most common occurring incidents were protamine reactions (1:783), coagulation problems (1:771), and heater/cooler failures 11:1809). The rate of occurrence of an incident resulting in a serious injury or death was one for every 1453 procedures. Although techniques and safety devices create a relatively secure environment for CPB, lower incident rates may be achieved with further improvements in coagulation monitoring and incident reporting. PMID- 10676869 TI - Vacuum-assisted venous return in pediatric cardiopulmonary bypass. AB - Vacuum-assisted venous return (VAVR) has been reported to offer benefits for adults undergoing cardiopulmonary bypass (CPB), such as improved venous return, lowering priming volume (by eliminating the need to prime the venous line), and the use of smaller venous cannulae. All these benefits would be of particular value in pediatric perfusion because of the unique challenges of these smaller patients and the relatively large components of the CPB circuit. We have been using VAVR in children since the early summer of 1998 after we became comfortable with the technique and convinced of its efficacy in adults. Ours is a medium sized pediatric caseload of slightly more than 100 CPB cases per year. With that caseload, it is most effective for us to minimize the inventory of different sizes of disposables used. We have opted for an oxygenator/reservoir that has a maximum flow of 4 liters with a priming volume of about 1 liter. We have been unhappy with the large prime volume in infants and earlier, in 1997-1998, were using a smaller prime oxygenator/reservoir until it was recalled. Faced again with a larger priming volume in the infants, we decided to try vacuum to decrease hemodilution and to evaluate other possible benefits. Through the use of VAVR, we have been able to decrease our priming volume, use smaller venous cannulae, and have more consistent return while experiencing no adverse effects of VAVR in our pediatric cardiac surgery patients. PMID- 10676870 TI - An in vitro evaluation of a new cannula tip design compared with two clinically established cannula-tip designs regarding aortic arch vessel perfusion characteristics. AB - We investigated in vitro aortic arch vessel perfusion characteristics of single and multiple jet-stream cannulae and a new dispersion stream tip aortic cannula. Pressures and flows of all arch vessels were measured while directing cannulae jets at the different arch vessels using 6 l/min pump flow. The highest increase in pressure above the set systemic level of 80 mmHg and increase in flow above the set normal flow distribution in the arch vessels occurred in the jet-streamed arch vessels with the single stream cannula. The values were as follows: 29 mmHg and 118 ml/min for the innominate artery, 28 mmHg and 42 ml/min for the left common carotid artery, and 25 mmHg and 54 ml/min for the left subclavian artery. The dispersion stream cannula showed increases in pressure and flow, followed by the multiple stream cannula. Aortic cannula tips and the orientation of jets are potential sources of imbalances of arch vessel perfusion with possible clinical implications regarding perfusion of arch vessels during extracorporeal circulation. PMID- 10676871 TI - Affinity pump system: a new peristaltic blood pump for cardiopulmonary bypass. AB - An in vitro study has been carried out to assess the pump performance of a new peristaltic, extracorporeal displacement pump (Affinity) for cardiopulmonary bypass. The pump system consists of a pump rotor (0-110 rpm), a pump chamber, a venous reservoir with a 5/8" connecting tube and the Affinity console. The polyurethane chamber is connected to the venous reservoir by a 5/8" tube and fills passively due to the hydrostatic pressure exhibited by the fluid height in the venous reservoir. The implementation of an occlusive segment in the pump chamber, which collapses in low filling states, should prevent significant negative pressures. An in vitro circuit was filled with bovine blood (37 degrees C, hematocrit 35%) and the pump flow was measured by an ultrasonic transit time flow probe with respect to pre-load, diameter and length of attached tubing in the venous line, pump speed (rpm) and size of the connecting tube (3/8" and 5/8"). At 108 rpm and a preload equal to 10 mmHg, the flow was 8.6 +/- 0.42 l/min for an afterload of 80 mmHg. The reduction of the inlet connector to 3/8" diminished the pump flow significantly to 5.2 +/- 0.31 l/min (p < 0.0001). The pump flow decreased linearly with respect to the length of the attached tube in the venous line and for a 2 m long 5/8" silicon tube, the rpm-optimized flow was still 6.0 +/- 0.28 l/min at a preload of 10 mmHg. In case of low filling state or too high rpm, the occlusive segment collapsed and no cavitation bubbles could be detected. Our in vitro measurements yield a nomogram for rpm-optimized blood flow with respect to the pre-load in the venous reservoir. The delivered 5/8"connecting tube facilitates optimum filling of the pump chamber for high blood flow, but limits the use of venous reservoirs to Affinity products. The pump yields a high blood flow even when long tubing in the venous line is used. This makes the pump a candidate for a ventricular assist device. In hypovolemia or high rpm, the occlusive segment collapses and no negative pressure is generated at the inflow site of the pump chamber. PMID- 10676872 TI - Effect of chronic paroxetine treatment on 5-HT1B and 5-HT1D autoreceptors in rat dorsal raphe nucleus. AB - This study reports the effect of chronic paroxetine (10 mg/kg p.o., 21 days) on 5 HT1B and 5-HT1D autoreceptors controlling stimulated 5-HT efflux in slices of rat dorsal raphe nucleus. Electrically evoked 5-HT (10 pulses, 200 Hz, 0.1 ms, 10 mA) was measured using fast cyclic voltammetry. 5-HT efflux was inhibited by CP 93129 (10 nM-10 microM) and by sumatriptan (1 nM-1 microM) agonists at 5-HT1B and 5 HT1D receptors, respectively. Chronic paroxetine did not, initially, appear to alter the sensitivity of the 5-HT1B autoreceptors to CP 93129. However, when constructed in the presence of WAY 100635 (10 nM) the selective and silent 5-HT1A antagonist, there was a significant (P < 0.001) rightward shift of the CP 93129 concentration-response curve in the paroxetine-treated rats but not in the controls, implying a desensitisation of the 5-HT1B autoreceptor by paroxetine. Chronic paroxetine did not affect the sumatriptan concentration-response curve, even with WAY 100635 present, implying that there was no (de)sensitisation of the 5-HT1D autoreceptor. These data suggest that chronic paroxetine treatment may desensitise 5-HT1B autoreceptors in the dorsal raphe nucleus but that this effect is unmasked only when the dominant 5-HT1A autoreceptor control is antagonised. PMID- 10676873 TI - The alpha-ketoglutarate dehydrogenase complex in neurodegeneration. AB - Altered energy metabolism is characteristic of many neurodegenerative disorders. Reductions in the key mitochondrial enzyme complex, the alpha-ketoglutarate dehydrogenase complex (KGDHC), occur in a number of neurodegenerative disorders including Alzheimer's Disease (AD). The reductions in KGDHC activity may be responsible for the decreases in brain metabolism, which occur in these disorders. KGDHC can be inactivated by several mechanisms, including the actions of free radicals (Reactive Oxygen Species, ROS). Other studies have associated specific forms of one of the genes encoding KGDHC (namely the DLST gene) with AD, Parkinson's disease, as well as other neurodegenerative diseases. Reductions in KGDHC activity can be plausibly linked to several aspects of brain dysfunction and neuropathology in a number of neurodegenerative diseases. Further studies are needed to assess mechanisms underlying the sensitivity of KGDHC to oxidative stress and the relation of KGDHC deficiency to selective vulnerability in neurodegenerative diseases. PMID- 10676874 TI - A critical histidine in the vesicular acetylcholine transporter. AB - The role of proton binding sites in the vesicular acetylcholine transporter was investigated by characterization of the pH dependence for the binding of [3H]vesamicol [(-)-trans-2-(4-phenylpiperidino)cyclohexanol] to Torpedo synaptic vesicles. A single proton binds to a site with pKa 7.1 +/- 0.1, which is characteristic of histidine, to competitively inhibit vesamicol binding. The histidine-selective reagent diethylpyrocarbonate causes time-dependent inhibition of [3H]vesamicol binding with a rate constant only about 20-fold lower than for reaction with free histidine. Because its pH titration has a simple, ideal shape, this residue probably controls all pH effects in the transporter between pH 6-8. Inhibition of [3H]vesamicol binding by diethylpyrocarbonate was slowed by vesamicol but not acetylcholine, which binds to a separate site. The data suggest that a critical histidine with a pKa of 7.1 is unhindered when reacting with diethylpyrocarbonate. A conformational model for the histidine is proposed to explain why acetylcholine competes with protons but not with diethylpyrocarbonate. A conserved histidine in transmembrane helix VIII possibly is the histidine detected here. PMID- 10676875 TI - Activation of apoptosis-linked caspase(s) in NMDA-injured brains in neonatal rats. AB - Unilateral injection of 50 nmol of N-methyl-D-aspartate (NMDA) into the left posterior striatum of 7 day-old rat pups induces massive neuronal loss in the ipsilateral hemisphere in 5 days. In this model of excitotoxicity, the form of neuronal death (necrosis vs apoptosis) has not been clearly addressed. Here we report evidence of DNA laddering in the ipsilateral hemisphere 24 h after the NMDA injection. Activation of apoptosis-linked caspase(s) was also identified, as evidenced by (i) the formation of caspase-produced 120 kDa alpha-spectrin breakdown product (SBDP120) and (ii) increase in hydrolysis of caspase-3 substrate acetyl-DEVD-7-amido-4-methylcoumarin in the homogenate from the ipsilateral hemisphere. Lastly, we note that i.p. injection (100 mg/kg) of a pan caspase inhibitor Z-D-DCB attenuates the levels of SBDP120. Our results suggest the presence of caspase-activation in this rat pup model of NMDA toxicity. PMID- 10676876 TI - Effect of ethanol on ATP-induced phospholipases C and D and serine base exchange in glioma C6 cells. AB - The effect of extracellular ATP, a nucleotide receptor agonist in the central nervous system, was investigated in glioma C6 cells on the intracellular Ca2+ level and the formation of phosphatidylethanol and phosphatidic acid in the presence and absence of ethanol (150 mM). In the cells prelabeled with [14C]palmitic acid, 100 microM ATP induced both the hydrolysis and the transphosphatidylation reactions leading to the formation of [14C]phosphatidic acid; addition of ethanol generated [14C]phosphatidylethanol. However, ATP mediated increase in the level of [14C]phosphatidic acid was not inhibited by ethanol. Furthermore, ethanol augmented ATP-induced transient and sustained increase in the intracellular Ca2+ concentration, whereas ethanol alone did not produce any change in the intracellular Ca2+ level. These results indicate that in glioma C6 cells, ATP induces activation of polyphosphoinositide-specific phospholipase C and phospholipase D and that ethanol enhances this effect. In the present investigation we have also shown that long-term (2 days) ethanol treatment, at concentration relevant to chronic alcoholism (100 mM), decreased the incorporation of [14C]serine into phosphatidylserine. Since the effect of ethanol on ATP-induced activities of phospholipase C and phospholipase D and on serine base-exchange in glioma C6 cells differs significantly from that in cultured neuronal cells, these results may contribute to a better understanding of the mechanisms of ethanol action in cells of glial origin. PMID- 10676877 TI - Psychotomimetics moderately affect dopamine receptor binding in the rat brain. AB - The hypothesis that psychotomimetics induce a rapid dopamine receptor regulation that could participate in the expression of the brain dopaminergic overactivation and in the early signs of psychotic-like behaviour, was checked by radioligand binding on rat brain cryosections. For this purpose, subchronic 7-day-d amphetamine pretreatment was combined with acute amphetamine, phencyclidine or LSD challenge. Acute application of psychotomimetics affected only striatal and accumbens but not nigral and olfactory dopamine receptor binding after 40 min, while subchronic amphetamine expressed no effect, as revealed by two-way ANOVA. Post-hoc statistical analysis showed that only striatal and accumbens[3H]SCH 23390 binding decrease (10-12%) following phencyclidine and striatal [3H]spiperone binding increase (11%) after acute amphetamine were significant. It is assumed that such moderate dopamine receptor binding changes probably reflect the fast receptor regulation responses without important influence on a proposed drug-induced dopaminergic overactivity. The registered alterations of D1 receptor binding after phencyclidine are suggested to be capable to modify the activity of some other neural pathways in the basal ganglia and thus participate in a psychotic-like behaviour. PMID- 10676878 TI - Expressions of amyloid precursor protein, synaptophysin and presenilin-1 in the different areas of the developing cerebellum of rat. AB - This study reveals the expressions of Alzheimer's disease-related amyloid precursor protein, presenilin-1, and a presynaptic marker protein, synaptophysin, in the archi-, paleo- and neocerebellum during the postnatal development of the rat. The Western blot results demonstrate a gradual increase in the soluble amyloid precursor protein level in the archicerebellum during the first 3 weeks, while in the neo- and paleocerebellum the levels reach a plateau as early as the 1st week. Immunohistochemically, the protein is present in the deep part of the external granule cell layer and the internal granule cell layer in the newborn animal, while in 3-week-old animals the staining appears mainly in the perikarya and dendrites of the Purkinje cells. The level of synaptophysin increases progressively from postnatal day 7 up to 3 weeks in the archi- and paleocerebellum, and up to 6 weeks in the neocerebellum. Immunohistochemically, the amyloid precursor protein staining appears first in the inner part of the molecular layer and in the internal granule cell layer. In a 3-week-old animal, synaptophysin staining is present in all areas of the cerebellar molecular layer and in the internal granule cell layer. The presenilin-1 immunohistochemical reaction appeared equally in the archi-, paleo- and neocerebellum. Much of the staining is present in the glial cells and Purkinje cells. Less immunoreactivity is observed in the Golgi cells and granule cells. It is concluded that the postnatal expressions of soluble and membrane-bound amyloid precursor protein, synaptophysin and presenilin-1 are regulated differently during the ontogenetical development of the archi-, paleo- and neocerebellum of rat. Further, the amyloid precursor protein and presenilin-1 may be present in cells which do not degenerate in Alzheimer's disease. PMID- 10676879 TI - Internalization and down-regulation of muscarinic acetylcholine receptors in cerebellar granule cells of tenascin-gene deficient mice. AB - The expression of tenascin-C on oligodendrocytes parallels the migration of granule cells in the developing cerebellum, indicating a role for tenascin-C as a guide for granule neurons to find their proper locations. In this study, cultured cerebellar granule neurons from tenascin-C-knockout mice were used to examine the role of tenascin-C in agonist-induced muscarinic acetylcholine receptor down regulation. Exposure of granule cells from wild-type or tenascin-C-negative mice to the muscarinic acetylcholine receptor agonist carbachol (1 mM) resulted in normal sequestration of cell-surface muscarinic acetylcholine receptors as assessed by [3H]N-methylscopolamine binding; however, down-regulation of total muscarinic acetylcholine receptors, measured with [3H]quinuclidinyl benzilate, was inhibited in granule cells from tenascin-C-negative mice. Remarkably, incubation of the tenascin-C-negative cells with the microtubule stabilizer taxol (10 microM) restored down-regulation of total muscarinic acetylcholine receptors to normal levels. We speculate that agonist-induced down-regulation of muscarinic acetylcholine receptors is functionally associated with tenascin-C-regulated microtubule structures in the developing cerebellum. PMID- 10676880 TI - Glutamate-mediated inhibition of oxidative phosphorylation in cultured retinal cells. AB - Glutamate is an excitotoxin responsible for causing neuronal damage associated with mitochondria dysfunction. We have analyzed the relationship between the mitochondrial respiratory rate, the membrane potential (delta psi) and the activity of mitochondrial complexes in retinal cells in culture, used as neuronal models. Glutamate (10 microM-10 mM) dose-dependently decreased the O2 consumption and the membrane potential. A linear correlation was found between these parameters, suggesting that the mitochondrial respiratory function was affected. Exposure to glutamate (100 microM) for 10 min, in the absence of Mg2+, inhibited the activity of complex I (26.3%), complexes II/III (22.2%) and complex IV (26.7%). MK-801 ((+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]-cyclohepten-5,10 imine hydrogen maleate), a non-competitive antagonist of the NMDA (N-methyl-D aspartate) receptors, completely reversed the effect exerted by 100 microM glutamate at the level of complexes I, II/III and IV. These results suggest that NMDA receptor-mediated inhibition of mitochondrial respiratory chain complexes may be responsible for the alteration in the respiratory rate of chick retinal cells submitted to glutamate. PMID- 10676881 TI - Transient treatments with L-glutamate and threo-beta-hydroxyaspartate induce swelling of rat cultured astrocytes. AB - We characterized swelling of rat cultured astrocytes induced by L-glutamate and its analogues. Among L-glutamate receptor agonists, L-glutamate, L-aspartate, L cysteic acid, DL-homocysteic acid, quisqualate and (+/-)-1-aminocyclopentane trans-1,3-dicarboxylic acid (trans-ACPD) increased astrocytic intracellular volume (3H-OMG space), while kainate, and N-methyl-D-aspartate did not. Threo beta-hydroxyaspartate (TBHA), D-aspartate and L-trans-pyrrolidine-2,4 dicarboxylic acid, high-affinity substrates for Na+-dependent L-glutamate transporters, increased astrocytic 3H-OMG space. L-Glutamate (0.5 mM) increased astrocytic 3H-OMG space to 300% of control in 40-60 min. The increase in 3H-OMG space by 1 mM TBHA was comparable to the L-glutamate-induced one. After a 10 min exposure to 0.5 mM L-glutamate, astrocytic 3H-OMG space was further increased to 200% even in the absence of L-glutamate. Astrocytes transiently exposed to L glutamate did not increase their cell volume in K+-free medium and in the presence of 1 mM ouabain, a Na+-K+ ATPase inhibitor. The increase after a transient exposure was also observed by a treatment of 1 mM TBHA, but not by 0.5 mM quisqualate. These results suggest that the volume increases after a transient treatment are mediated by activation of Na+-dependent L-glutamate transporter. PMID- 10676882 TI - Purification and embryotropic roles of tissue inhibitor of metalloproteinase-1 in development of "HanWoo" (Bos taurus coreanae) oocytes co-cultured with bovine oviduct epithelial cells. AB - This study was conducted to purify a tissue inhibitor of metalloproteinase (TIMP) 1 in a serum-free medium conditioned with bovine oviduct epithelial cells (BOEC) and to evaluate its effect on development of "HanWoo" (Bos taurus coreanae) embryos to the blastocyst stage. In the first study using SDS-PAGE electrophoresis, the presence of 32 kDa proteins, which contains TIMP-1, was detected in the medium conditioned with BOEC, and TIMP-1 was then purified from the medium by gel filtration and HPLC techniques. When examined TIMP-1 secretion, fluorescent foci indicating the secretion of TIMP-1 were found after stained BOEC with fluorescein isothiocyanate. In the next experiment, two-cell embryos derived from in vitro-fertilization were cultured in a serum-free medium, to which 0, 1.25, 2.5 or 5 microg/ml of purified TIMP-1 was supplemented. More (P<0.05) embryos developed to the morula and blastocyst stages after the addition of 2.5 microg/ml to culture medium than after no addition. In conclusion, our data indicate that BOEC secrete TIMP-1 and this glycoprotein promotes the prehatched development of "HanWoo" embryos derived from in vitro-fertilization. PMID- 10676883 TI - Adaptation of the muscles of mastication to the flat skull feature in the polar bear (Ursus maritimus). AB - The muscles of mastication of the polar bear (Ursus maritimus) and those of the brown bear (U. arctos) were examined by anatomical approach. In addition, the examination of the skull was carried out in the polar bear, brown bear and giant panda (Ailuropoda melanoleuca). In the polar bear, the rostro-ventral part of the superficial layer of the M. masseter possessed the abundant fleshy portion folded in the rostral and lateral directions like an accordion. Moreover, the rostro medial area of the superficial layer became hollow in the nuchal direction when the mouth was closed. The M. temporalis of the polar bear covered up the anterior border of the coronoid process of the mandible and occupied the almost entire area of the cranial surface. The M. pterygoideus medialis of the polar bear was inserted on the ventral border of the mandible and on the ventral part of the temporal bone more widely than that of the brown bear. As results of our measurements of the mandible, an effect of the leverage in the polar bear was the smallest in three species. In the polar bear, the skull was flat, and the space between zygomatic arch and ventral border of the mandible, occupied by the M. masseter was the narrowest. It is suggested that the muscles of mastication of the polar bear is adapted to the flat skull feature for supplementing the functions. PMID- 10676884 TI - Synthesis and secretion of A-type natriuretic peptide in the auricular cardiocytes during pregnancy and lactation in mouse. AB - A-type (atrial) natriuretic peptide (ANP) levels in the auricular cardiocytes and plasma were examined by immunohistochemistry, electron microscopy, and radioimmunoassay in pregnant and lactating mice. Additionally, the cardiocyte ANP mRNA expression was measured by the polymerase chain reaction method. ANP immunoreactivity (IR) and the number of ANP-granules in the cardiocytes on the 18th day of gestation were greater than those in virgin controls, but the plasma ANP concentration decreased on the 18th day of gestation. On the day of delivery, ANP-IR and the number of ANP-granules in the cardiocytes were decreased compared to those during the pregnancy and to those in virgin controls, and then began to increase continually until the 15th day of lactation. Plasma ANP concentration after delivery was significantly higher than that during pregnancy, and than that in virgin controls, and continued to increase until the 15th day of lactation. Cardiocyte ANP mRNA expression was highest on the day of delivery compared to that in all the other times. In conclusion, these results suggested that the circulating systems of ANP during pregnancy and lactation were regulated differentially. PMID- 10676885 TI - Apoptosis and cell proliferation in rat hepatocytes induced by barbiturates. AB - To examine the effect on cell population in hepatocytes of phenobarbital (PB) and other barbiturates, PB, allobarbital (ALB), barbital sodium (BS) and barbituric acid (BA) were given orally to male rats for 7 consecutive days. Although there was no apparent change in non-promoting BA, hepatomegaly was induced by PB, BS and ALB, which are promoters of hepatocarcinogenesis. In PB- and BS-treated livers, hepatomegaly was attributable to hepatocyte proliferation and enzyme induction. In ALB-treated liver, it was attributable to enzyme induction. The level of cell proliferation was reduced to less than the control values following withdrawal of PB, ALB and BS. It seemed that the degree of suppression of cell proliferation following withdrawal of these compounds correlated to the degree of cell proliferation (PB>BS>ALB) during treatment. In PB-treated liver, apoptosis was induced during treatment, serving to eliminate the excess of hepatocytes. This suggests that short-term administration of PB neither induced suppression of apoptosis nor disturbed homeostasis of hepatocyte populations. PMID- 10676886 TI - Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice. AB - MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3+ CD4- CD8- B220+ (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules, CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice. PMID- 10676887 TI - Detection of annexins I and IV in bronchoalveolar lavage fluids from calves inoculated with bovine herpes virus-1. AB - Annexins are phospholipid-binding proteins and are abundant in the lung. Annexins I and IV, but not II and VI, have been detected in bronchoalveolar lavage (BAL) fluids from calves inoculated with Pasteurella haemolytica, the pathogen for calf pneumonia. In this study, BAL fluids from calves with experimental pneumonia induced by inoculation to right lung lobes of bovine herpes virus-1 (BHV-1), the major viral pathogen for pneumonia, were examined for detection of annexins I and IV. Of 6 calves inoculated with BHV-1, annexins I and IV were coincidentally detected in BAL fluids from right lung lobes of 4 calves, but not in BAL fluids from left lung lobes of 6 inoculated calves or those from left and right lung lobes of 3 control calves. Annexin II and VI were not found in any BAL fluids examined. These results, together with previous findings on calves inoculated with Pasteurella haemolytica, suggest that the release of annexins I and IV onto the alveolar surface is an essential event occurring in response to pulmonary infections of BHIV-1 and Pasteurella haemolytica. PMID- 10676888 TI - Effects of antioxidants on induction of apoptosis in bursal cells of Fabricius during in vitro cultivation. AB - After physically disrupting cell contacts, apoptosis of bursal cells of Fabricius was induced during in vitro cultivation. The percentage of apoptotic cells increased with incubation time and approximately 70% cells represented apoptosis after 6 hr of incubation. The induction of apoptosis was significantly inhibited by treatment of the cells with ascorbic acid (vitamin C), but not with trolox, a vitamin E analog. An intense DNA ladder pattern was shown at 6 hr post-isolation, which is a biochemical hallmark of apoptosis. Treatment of the cells with ascorbic acid inhibited the DNA fragmentation, but trolox did not. To monitor the intracellular production of reactive oxygen species (ROSs), the intensity of fluorescence emitted from DCFH-DA was measured. The intensity of fluorescence from cells incubated for 0.5-2 hr was approximately 2-fold higher than that from cells at 0 hr. The relative intensity of fluorescence decreased immediately after the addition of ascorbic acid to the cells. The intensity from the cells treated with ascorbic acid was 20-30% of that from the control cells at each incubation time. For trolox, the intensity was 50-70% of that from the control cells at each 1 to 2 hr incubation time. When ROSs-induced lipid peroxidation was assessed using cis-parinaric acid (PnA) as a monitor molecule, lipid peroxidation was found to occur in the control cells after isolation of the bursal cells. Treatment of the cells with trolox reduced lipid peroxidation, but treatment with ascorbic acid enhanced peroxidation. PMID- 10676889 TI - Decreased apolipoprotein C-III concentration in the high-density lipoprotein fraction from calves inoculated with Pasteurella haemolytica and bovine herpes virus-1. AB - Lipoprotein lipid and apoprotein concentrations are known to be altered during the acute-phase response. We have previously shown that the serum activity of lecithin:cholesterol acyltransferase (LCAT) and concentration of cholesteryl esters, both constituents of the high-density lipoprotein (HDL) fraction, are reduced in calves inoculated with Pasteurella haemolytica and bovine herpes virus 1, the two major pathogens for calf pneumonia. The concentration of apolipoprotein C-III (apoC-III), a low molecular mass protein component distributed mainly in the HDL fraction, was therefore examined in bacteria- and virus-inoculated calves. An enzyme-linked immunosorbent assay demonstrated that it was decreased by inoculations of Pasteurella haemolytica and bovine herpes virus-1. The decrease was detected as early as 1 day after inoculation in both groups. A decreased serum apoC-III concentration was also observed by immunoblot analysis. It was detected in the HDL fractions from the bacteria- and virus inoculated calves, and HDL apoC-III concentrations in the inoculated calves were decreased compared with controls. These results, coupled with the previous findings on LCAT activity and the cholesteryl ester concentration, indicate that a decreased HDL concentration is one of the early events occurring during the acute-phase response evoked by infections with Pasteurella haemolytica and bovine herpes virus-1. PMID- 10676890 TI - Regional specialization of ganglion cell layer of the chick retina. AB - Specialization of the ganglion cell layer (GCL) was studied by Nissl-staining and axonal tract-tracing methods in chicks and chick embryos. The changes in the retinal area and the cell number in the GCL produced a disparity in the cell density that occurred through the two different processes, cell generation (before embryonic days 10-14, E10-14) and cell loss (after E10-14). One high density area was found in the retinal fundus on E8 (presumptive central area, pCA) and its density decreased toward the peripheral retina. Another high-density area was found in the dorsal retina on E11 (presumptive dorsal area, pDA). Cell densities of the pCA and the pDA on E11 decreased gradually to 25-30% by P1, and after that they further decreased to 40-60% by P30. The pCA was still identified on P30, but the pDA became very obscure by this age. In contrast, ganglion cell sizes increased 5-7 times in the pCA and pDA from E8 to P30, and increased 12 times in the temporal periphery. The present study suggests that the center peripheral gradient of cell density results from lager scale of cell genesis in the pCA, but not from lager scale of cell loss in the peripheral retina. However, obscuration of the pDA results from equalization of cell density in cellular degeneration processes. PMID- 10676891 TI - A hematological study on thirteen cats with myelodysplastic syndrome. AB - Hematological abnormalities were investigated in 13 cats with myelodysplastic syndrome (MDS). Examination of the peripheral blood samples from the 13 cats revealed anemia in 11 cats, leukopenia in 9 cats, and thrombocytopenia in 9 cats. Four cats had pancytopenia (30.8%) and 9 cats had bicytopenia (69.2%). Dysplastic changes of erythrocytes, neutrophils, and platelets in the peripheral blood were found in 5, 10 and 8 cats, respectively. Bone marrow examination of the 13 cats revealed that ratios of blast cells to all nucleated cells (ANC) ranged from 0 to 20%. Ratios of erythroid progenitor cells to ANC were more than 50% in 3 cats and less than 50% in 10 cats. Eosinophils accounted for more than 5% of non-erythroid cells in 10 cats. Dysplastic changes in the granurocytic, erythrocytic, and megakaryocytic cells in the bone marrow were found in 11, 7 and 5 cats, respectively. Dysplastic changes in these cats included giant neutrophils, ring nucleated neutrophils, binuclear myelocytes, hypersegmented and hyposegmented neutrophils, megaloblastoid erythroblasts, multinucleated erythroblasts, micromegakaryocytes, and segmented multinucleated megakaryocytes. Virological examination indicated the presence of feline leukemia virus antigen in the peripheral blood from all of the 13 cats with MDS. The peripheral blood cytopenias and dysplastic changes in each blood cell lineage in the bone marrow were shown to be important for the diagnosis of MDS in cats. PMID- 10676892 TI - Induction of dog sperm capacitation by glycosaminoglycans and glycosaminoglycan amounts of oviductal and uterine fluids in bitches. AB - Ejaculated sperm collected from 12 beagle dogs were incubated in canine capacitation medium (CCM), supplemented with 5 microg/ml chondroitin sulfate A (CS), 5 microg/ml hyaluronic acid (HA), or 5 microg/ml heparin (HP) for 7 hr at 38 degrees C in a 5% CO2 in air atmosphere to investigate the effects of glycosaminoglycans (GAGs) on dog sperm capacitation. The percentages of motile sperm, hyperactivated sperm (%HY), and acrosome-reacted sperm (%AR) in all media were examined after 4 hr and 7 hr of incubation. The oviducts and uteri of 9 anestrous and 18 estrous beagle bitches were removed under halothane inhalation anesthesia to measure the total GAG amounts in oviductal and uterine fluids. The lumens of the ampulla of the oviducts, isthmus of the oviducts, and the uterine horns were each flushed with 1 ml HEPES-EDTA fluid. Total GAG amounts in the flush fluids obtained were measured with a spectrophotometer. Sperm motility (51 59%), %HY (79-86%), and %AR (31-36%) in CCM supplemented with CS, HA, or HP were significantly higher after 7 hr of incubation than when incubated in CCM without GAGs (P<0.01 or 0.05). The mean total GAG amounts in the fluids from the ampulla and isthmus of the oviducts and the uterine horns in the estrous bitches were higher than in the anestrous bitches. These results indicate that GAGs in the oviductal and uterine fluids in estrous bitches are associated with in vivo sperm capacitation. PMID- 10676893 TI - Effects of orchidectomy on bone metabolism in beagle dogs. AB - The effects of orchidectomy on bone metabolism in male beagle dogs were examined using twelve 2-year-old dogs that were orchidectomized. The dogs' bilateral iliac bones, double-labeled with tetracycline and calcein for the histomorphometry, were obtained from three dogs prior to orchidectomy and at 3, 6, 9, and 12 months afterwards. The serum biochemical constituents related to bone metabolism were examined before and every month after orchidectomy. Between 1 and 6 months after orchidectomy, the value of serum testosterone decreased (1 month), while the levels of parathyroid hormone, calcitonin, total calcium, osteocalcin, and alkaline phosphatase activity increased significantly, indicating a high bone turnover. The mean trabecular thickness and the fraction of labeled osteoid surface decreased significantly 3 months after orchidectomy, but other histomorphometric parameters were unchanged. In the period 7-12 months after orchidectomy, the parathyroid hormone level increased ever and above that of the first 6-month period, while the levels of calcitonin, osteocalcin, alkaline phosphatase activity, and phosphorus decreased. The bone volume, mean trabecular thickness, and the fraction of labeled trabecular surface decreased significantly compared with the pre-orchidectomy values. These findings indicate an imbalance in bone metabolism (i.e. bone resorption > bone formation). These results indicate that a loss of bone volume accompanied the fall in sex hormone levels following orchidectomy and suggest that the orchidectomized dog is available as an animal model for studying osteoporosis caused by hypogonadism and the decline of sex functions in men. PMID- 10676894 TI - 1Alpha-hydroxyvitamin D3 prevents the decrease of bone mineral density in lactating beagles. AB - We assessed the change of bone mineral density (BMD) in lactating beagles with dual energy X-ray absorptiometry (DXA) and the preventive effect of 1alpha hydroxyvitamin D3 (1alpha(OH)D3) on the BMD. Beagles, two to five years old, were used for detecting the time course change of BMD. Since the coefficient of variation (CV(%)) on detecting lumber vertebral (L2-L4) and tibial BMD by DXA was about 0.5%, DXA was useful to detect the change of BMD in beagles. There was a marked decrease in vertebral BMD during lactational period in the control group. The BMD levels after weaning were found to reverse to the initial level at mating. The same tendency was observed in tibial BMD as vertebral BMD, though the BMD changes were not marked. Beagles were administered at a dose of 0.1 microg/kg of 1alpha(OH)D3 three times in a week, and it was found to suppress the decrease in vertebral BMD during the breast feeding period. Also, the administration of 1alpha(OH)D3 promoted the prevention of decreased BMD during lactation both in vertebrae and tibiae. Significant effects of 1alpha(OH)D3 administration on tibial BMD were not observed. No adverse effects, such as hypercalcemia and hypercalciuria, were observed during the experimental period. Therefore, DXA was useful for detecting the changes of BMD in lactating beagles and the change of BMD was marked in lumber vertebrae, which are rich in trabecular bone. The preventive effect of 1alpha(OH)D3 on the decrease of BMD during the lactation period was observed in beagles. PMID- 10676895 TI - Antemortem evaluation for magnetic resonance imaging of the equine flexor tendon. AB - In this study antemortem evaluation of equine flexor tendons--the superficial digital flexor tendon and the deep digital flexor tendon--using magnetic resonance (MR) images was performed. Postmortem flexor tendons were used to prepare the slice positions, coil and body positions for MR imaging. It was possible by this method to take antemortem MR images of equine limbs that distinguished features as well as postmortem images described in previous studies. The total time of antemortem scanning was about 40 min. This study is the first to report antemortem MR images in horses. PMID- 10676896 TI - Effects of chlorine, iodine, and quaternary ammonium compound disinfectants on several exotic disease viruses. AB - The effects of three representative disinfectants, chlorine (sodium hypochlorite), iodine (potassium tetraglicine triiodide), and quaternary ammonium compound (didecyldimethylammonium chloride), on several exotic disease viruses were examined. The viruses used were four enveloped viruses (vesicular stomatitis virus, African swine fever virus, equine viral arteritis virus, and porcine reproductive and respiratory syndrome virus) and two non-enveloped viruses (swine vesicular disease virus (SVDV) and African horse sickness virus (AHSV)). Chlorine was effective against all viruses except SVDV at concentrations of 0.03% to 0.0075%, and a dose response was observed. Iodine was very effective against all viruses at concentrations of 0.015% to 0.0075%, but a dose response was not observed. Quaternary ammonium compound was very effective in low concentration of 0.003% against four enveloped viruses and AHSV, but it was only effective against SVDV with 0.05% NaOH. Electron microscopic observation revealed the probable mechanism of each disinfectant. Chlorine caused complete degeneration of the viral particles and also destroyed the nucleic acid of the viruses. Iodine destroyed mainly the inner components including nucleic acid of the viruses. Quaternary ammonium compound induced detachment of the envelope of the enveloped viruses and formation of micelle in non-enveloped viruses. According to these results, chlorine and iodine disinfectants were quite effective against most of the viruses used at adequately high concentration. The effective concentration of quaternary ammonium compound was the lowest among the disinfectants examined. PMID- 10676897 TI - Hematological and plasma biochemical values in captive Eld's-Brow Antlered deer (Cervus eldi thamin) in Thailand. AB - Blood samples were collected from 20 sedated captive Eld's-Brow Antlered deer (Cervus eldi thamin), aged over 1.5 years, to define their mean hematological values (packed cell volume and hemoglobin) and mean plasma biochemical parameters. Male deer had a significantly higher plasma glucose level and aspartate aminotransferase activity than female deer. PMID- 10676898 TI - Sequence analysis of the genes encoding the phosphoprotein of recent isolates of canine distemper virus in Japan. AB - The nucleotide sequences of the phosphoprotein (P) of canine distemper virus (CDV) strains isolated between 1992 and 1996 in Japan were determined. This is the first report of the complete sequences of the P genes of recently prevalent CDV strains. The deduced amino acid sequences of the P, C and V proteins showed that in the new Japanese isolates, these proteins have approximately 93%, 90-91% and 92% identities with those of the Onderstepoort vaccine strain, respectively. The predicted functional regions were conserved. RNA editing resulting in a shift to the open reading frame (ORF) of the V protein was shown to occur with the same efficiency in both the field isolates and vaccine strain. PMID- 10676899 TI - Surgical repair of a depressed fracture in a green sea turtle, Chelonia mydas. AB - Sea turtles are considered to be endangered species. A depressed fracture of a 35 kg green sea turtle was treated surgically. Isoflurane was used for induction and maintenance of anesthesia. Slow induction of and slow recovery from anesthesia was remarkable. After the operation, there was an improvement of general status, but head tilt and weakness of the left limbs persisted. As the turtle did not eat, force feeding using stomach tube was performed. The turtle died at about 6 months after the surgery. PMID- 10676900 TI - Experimental reinfection with homologous porcine reproductive and respiratory syndrome virus in SPF pigs. AB - The present examination was conducted to determine if the pigs infected with one strain of porcine reproductive and respiratory syndrome virus (PRRSV) would be protected against a subsequent homologous virus challenge. Sixteen 4-week-old SPF pigs were assigned to 2 experimental groups A and B. The pigs in group A were inoculated with 10(6.5) TCID50 of PRRSV by intranasal route. On 77 days post inoculation (PI), pigs in groups A and B were similarly inoculated with same virus. After the secondary inoculation, the pigs in group A didn't show any clinical sign including pyrexia and reduction of white blood cell (WBC) number. Viremia was detected only on 3 days PI with low virus titer and any virus was not recovered from serum and tissues at the time of necropsy on 14 or 28 days PI. In contrast, pigs in group B showed pyrexia for 14 days and reduction of WBC number on 3 days PI. Viremia was detected between 3 and 28 days PI, and virus was isolated from several tissues of all pigs. These results indicate that previous exposure to PRRSV can prevent development of clinical signs and reduce virus proliferation in pigs after subsequent infection with the homologous PRRSV. PMID- 10676901 TI - Examination of quantitative analysis and measurement of the regurgitation rate in mitral valve regurgitation by the "proximal isovelocity surface area" method. AB - In 33 dogs with mitral valve insufficiency (MR), assessed as severe by semi quantitative color flow Doppler echocardiography, regurgitation volumes were measured by the "Proximal Isovelocity Surface Area" (PISA) method. Good correlation (p<0.01, r=0.97) between the regurgitation volumes determined by the "PISA" and pulsed Doppler methods was confirmed. As evaluated by the "PISA" method, regurgitation rates in the 32 dogs with measurable regurgitation volumes ranged from 23 to 73%, with a mean of 51.6 +/- 11.8%. Regurgitation volumes ranged from 3.3 to 32 ml, with a mean of 8.4 +/- 6.4 ml. PMID- 10676902 TI - Bone marrow necrosis in a cat infected with feline leukemia virus. AB - A one-year old castrated male cat was admitted to the hospital with vomiting and diarrhea. Laboratory examination revealed pancytopenia and positive for FeLV antigen. A bone marrow examination indicated necrosis of the nucleated cells. Based on these findings, the cat was diagnosed as bone marrow necrosis. Pancytopenia was effectively treated with corticosteroids. Re-examination of the bone marrow confirmed a recovery of normal hematopoietic cells with a infiltration of many macrophages. It is strongly suspected that the bone marrow necrosis in this case could be associated with a bone marrow suppression due to FeLV infection. PMID- 10676903 TI - Effects of perineural capsaicin treatment on compound action potentials of superior laryngeal nerve afferents in sevoflurane-anesthetized dogs. AB - Effects of perineural capsaicin (CAPS) treatment on compound action potentials of the superior laryngeal nerve (SLN) afferents were studied in 6 sevoflurane anesthetized dogs. Perineural CAPS (100 microg/ml) to the bilateral SLNs reduced (P<0.01) the peak and integral amplitudes of the C-wave of the compound action potential. By contrast, the perineural CAPS had no effect on the A-wave component (P>0.05). Removal of the perineural CAPS recovered the C-wave to pretreatment level. The perineural CAPS treatment selectively blocks C-wave compound action potentials of the SLN afferents, providing a useful tool for studies of laryngeal C-fibers in respiratory physiology. PMID- 10676904 TI - Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids. AB - Bovine in vitro matured and fertilized oocytes were cultured for 153 hr in groups of 3 or 30 in 30 microl of modified synthetic oviduct fluid medium supplemented with amino acids. The concentration of ammonium in culture medium at 153 hr of culture was significantly decreased by medium change at 72 hr of culture. However, regardless of embryo density, medium change had no beneficial or detrimental effect on the development of bovine embryos. Increase in the development to blastocysts and production of ammonium were observed when embryos were cultured in groups of 30. These results indicated that the ammonium concentration detected in this culture system has a negligible effect on the development of bovine embryos to blastocysts. PMID- 10676905 TI - Melanotic neurofibroma in a steer. AB - A melanotic neurofibroma in a steer was investigated histologically, immunohistochemically and ultrastructurally. A very large tumor mass was located in the region of the head and right cheek. The tumor tissue consisted of an admixture of cells resembling Schwann cells and spindle-shaped cells, and they frequently contained melanin granules. Neoplastic Schwann cells were positive for S100 protein, with variation in intensity of staining, but most spindled cells were S100 negative. The tumor cells displayed ultrastructural features similar to those of Schwann cells or perineurial cells. The presence of melanosomes in varying stages of melanization in both cell types suggests that they have a common origin. This is a tumor of neural crest origin showing schwannian and perineurial differentiation, with ectopic production of melanin granules. PMID- 10676906 TI - Introduction: acquired aplastic anemia. PMID- 10676907 TI - Hematopoietic cell destruction by immune mechanisms in acquired aplastic anemia. PMID- 10676908 TI - Myelodysplastic syndrome and aplastic anemia: distinct entities or diseases linked by a common pathophysiology? AB - It is often difficult to distinguish myelodysplastic syndrome (MDS) from severe aplastic anemia (SAA) because both can present with profoundly hypocellular bone marrows. The distinction matters because although both conditions are complicated by pancytopenia, the risk of progression to acute leukemia is much greater in MDS. This chapter reexamines the relationship between SAA and MDS. The clinical and morphological features and pathophysiology of AA (including moderate and severe forms of acquired AA) are compared with MDS and hypoplastic MDS, with particular reference to new observations implicating autoimmune processes in both conditions. SAA and hypoplastic MDS (HMDS) are discussed in the light of these findings and attempts to separate nonevolving bone marrow failure syndromes from marrow failure progressing to acute leukemia are reviewed. The weight of evidence supports a common pathophysiology and, more speculatively, a common etiology for at least some forms of AA and MDS. PMID- 10676909 TI - Current status of allogeneic bone marrow transplantation in acquired aplastic anemia. AB - Bone marrow transplantation is an effective therapy for aplastic anemia. Infusion of allogeneic hematopoietic stem cells after high-dose immune suppression restores normal hematopoiesis in most patients and long-term follow-up has confirmed the durability of donor hematopoiesis. However, success of this approach is limited by transplant-related complications, such as graft failure, graft-versus-host disease, and various organ toxicities. Long-term survival rates range from less than 40% to more than 90% in reported series. These rates have improved over the past 20 years due to significant reductions in graft-versus host disease, interstitial pneumonitis, and early transplant-related mortality. Most long-term survivors have excellent performance status. Late effects such as cataracts, thyroid disorders, joint problems, and therapy-related cancers are observed, especially in patients who received radiation for pretransplant conditioning. Results are best in young patients transplanted with bone marrow from a human leukocyte antigen (HLA)-identical sibling; early transplantation is appropriate in this group. For older patients or those without an HLA-identical related donor, transplants are better reserved for those who fail to respond to immunosuppressive therapy. PMID- 10676910 TI - Alternative-donor hematopoietic stem-cell transplantation for severe aplastic anemia. AB - Bone marrow transplantation (BMT) from a human leukocyte antigen (HLA)-matched sibling is the treatment of choice in children and young adults with severe aplastic anemia (SAA). However, because only 30% of patients have a suitable donor, more aggressive nontransplant immunosuppressive regimens have been used, with reasonable results. The use of mismatched family member and unrelated donors, initially fraught with problems of nonengraftment and severe graft-versus host disease (GVHD), has improved markedly over the past 10 years. The establishment of donor registries, more precise HLA typing methods, and better supportive care are significant factors in the improved outcomes. The challenge for the future is to assemble the optimal combination of donor selection, conditioning regimen, and GVHD prophylaxis to enhance disease-free survival. These better outcomes should encourage the treating physician to consider stem cell transplant at an earlier stage of disease. PMID- 10676911 TI - Immunosuppressive treatment of aplastic anemia with antithymocyte globulin and cyclosporine. AB - Immunosuppression is the treatment modality for the majority of patients with aplastic anemia, most of whom are not candidates for allogeneic stem-cell transplantation. Antithymocyte globulin (ATG) or antilymphocyte globulin (ALG) have proven to be essential components of all regimens. Initial response rates can be improved by the addition of cyclosporine A (CsA), and this combination has become the standard of care for appropriate patients. Several new approaches to immunosuppression are being studied, including the optimal timing of administration of these drugs, the use of novel immunosuppressive agents, and the addition of early- and late-acting hematopoietic growth factors. PMID- 10676912 TI - Treatment of acquired severe aplastic anemia: bone marrow transplantation compared with immunosuppressive therapy--The European Group for Blood and Marrow Transplantation experience. AB - Patients with severe aplastic anemia (SAA) can be successfully treated with bone marrow transplantation (BMT) or immunosuppressive therapy (IS). The current outcome using both forms of therapy among 3,669 patients treated in Europe between 1976 and 1998 is reviewed. Significant progress has been made and the overall risk of failure is now low, with survival rates greater than 80% for both treatments. Chronic graft-versus-host disease (GvHD) remains a problem for BMT patients, and carries a high risk of lethal complications. On the other hand, IS patients are exposed to late failure due to relapse or clonal/malignant diseases. First-line BMT from identical siblings is compared with IS therapy in an intent to-treat analysis of 1,765 patients, regardless of subsequent transplant status. The outcome of SAA patients has improved considerably over time and is influenced by patient variables such as severity of the disease and age, but also by the choice of the initial treatment. PMID- 10676913 TI - Hematopoietic growth factors in the pathogenesis and for the treatment of aplastic anemia. AB - The production and release of hematopoietic growth factors from bone marrow stromas established in vitro from patients with aplastic anemia is normal or increased. Addition of hematopoietic growth factors to aplastic anemia bone marrow cells results in only modest increases in colony growth, with the exception of granulocyte colony-stimulating factor (G-CSF), which corrects their impaired cloning efficiency to normal. Most clinical data on the use of hematopoietic growth factors in aplastic anemia have derived from uncontrolled and small single-arm studies or case reports. Sustained trilineage hematologic responses have not been observed when hematopoietic growth factors have been used alone or in combination. Serious side effects have been reported for most of the hematopoietic growth factors in patients with aplastic anemia, with the exception of G-CSF. There is a major concern that they may further increase the risk of clonal disorders such as myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Hematopoietic growth factors should not be used alone in newly diagnosed patients as specific treatment for aplastic anemia, and their use in combination with immunosuppressive therapy should be confined to multicenter, prospective randomized studies. PMID- 10676914 TI - Late clonal diseases of treated aplastic anemia. AB - Recent progress in the treatment of aplastic anemia has dramatically changed the previously grim prognosis for these patients. Improvements in bone marrow transplantation and immunosuppression have increased the number of long-term survivors so that immediate survival is no longer the sole concern. Here, we review the major clinical studies and summarize recent analyses of risk factors for developing paroxysmal nocturnal hemoglobinuria (PNH), myelodysplastic syndrome (MDS), acute leukemia, or solid tumor after treatment for aplastic anemia. We also examine biologic clues that may shed light on the interrelationship between aplastic anemia and clonal diseases. PMID- 10676915 TI - Cyclophosphamide and other new agents for the treatment of severe aplastic anemia. AB - Severe aplastic anemia (SAA) has a poor prognosis in the absence of treatment. Current accepted therapeutic strategies include allogeneic stem-cell transplantation and immunosuppression, both resulting in long-term survival in the majority of patients. Although human leukocyte antigen (HLA)-matched sibling stem-cell transplantation is highly effective, the 25% probability of finding a suitable sibling donor within a family renders this approach available to only a minority of patients. Transplantation using HLA-matched, unrelated donors carries a high risk of treatment failure along with considerable toxicity. While combined immunosuppression with both antithymocyte globulin (ATG) and cyclosporine A (CSA) produces hematologic improvement in most patients, relapse is common. Late evolution of aplastic anemia to other serious hematologic disorders, including paroxysmal nocturnal hemoglobinuria (PNH), myelodysplasia, and acute leukemia, is also a significant problem following treatment with ATG/CSA. Recently, results of immunosuppression in SAA with another potent immunosuppressive agent, cyclophosphamide, were reported in a small number of patients. The overall response rate was similar to that seen with ATG/CSA, but relapse and late clonal disease were not observed during a long period of follow-up. A larger randomized trial comparing sustained hematologic response rates to either conventional immunosuppression with ATG/CSA or high-dose cyclophosphamide and CSA is now underway; secondary end points include response duration, event-free survival, and overall survival. Additionally, a number of protocols designed to test the efficacy of alternative immunosuppressive or immunomodulatory agents are being developed. PMID- 10676916 TI - Splenectomy-sparing strategies for the treatment and long-term maintenance of chronic idiopathic (immune) thrombocytopenic purpura. AB - Patients with idiopathic thrombocytopenic purpura (ITP) have vulnerability to additional bleeding, leaving them susceptible to severe hemorrhaging. Low platelet counts contribute to this rare, but significant outcome, but may not be the sole determinant. Although the only current treatment of ITP felt to be curative is surgical removal of the spleen, the long-term outcome for these patients is not well defined. Our group Investigated the use of Intravenous gamma globulin in the treatment of children with chronic ITP as a means to defer splenectomy. A variation of this approach uses anti-D to block splenic macrophages with antibody-coated red blood cells. There may be a correlation between response to anti-D and response to splenectomy in adults with ITP. Because the long-term results of splenectomy are not well defined, additional clinical studies are warranted. Questions requiring further study Include whether repeated Infusions of anti-D could allow the postponement and ultimate avoidance of splenectomy and whether the role of anti-D may be for pregnant women who are not readily eligible for splenectomy. Such an analysis should include information on the long-term outcome of splenectomy as well as information on whether patients have a durable improvement. Such studies about the potential outcomes of splenectomy and of avoidance of splenectomy will help identify new treatment strategies that may help to eliminate the need for this procedure in patients with chronic ITP. PMID- 10676917 TI - The pathogenesis of chronic immune (idiopathic) thrombocytopenic purpura. AB - Chronic Immune (idiopathic) thrombocytopenic purpura (ITP) is an autoimmune disorder in which antiplatelet autoantibody induces platelet destruction. Platelet surface membrane proteins become antigenic, stimulating the immune system to produce autoantibody. The initial antigenic response probably occurs in the spleen, inducing autoantibody production followed by stimulation of other antibody-producing tissues, particularly the bone marrow. Autoantibodies against either platelet glycoprotein (GP)IIb/IIIa or GPIb/IX are produced by about 75% of ITP patients and can be detected using antigen-specific assays. The spleen is the major site of platelet destruction in ITP because of its unique milieu. About one third of the platelet mass is present in the spleen at all times, where the local production of antiplatelet antibody leads to high autoantibody concentrations. These antibody-sensitized platelets circulate slowly through the phagocytic cell rich spleen, resulting in their destruction. Since autoantibody binds to both platelets and megakaryocytes, both platelet destruction and inhibition of thrombopoiesis may be of importance in the pathogenesis of chronic ITP. PMID- 10676918 TI - The spleen and splenectomy in immune (idiopathic) thrombocytopenic purpura. AB - The benefits of surgical splenectomy in patients with immune (Idiopathic) thrombocytopenia purpura (ITP) probably reflect the combined effects of eliminating a source of antiplatelet antibody synthesis as well as the primary site of platelet destruction. The recent availability of intravenous Rho(D) Immune globulin (WinRho SDF; Nabi, Boca Raton, FL) presents an opportunity to extend the duration of nonsurgical (spleen-sparing) management of chronic ITP by inducing reversible Fc blockade. While new methods for laparoscopic splenectomy may offer improved surgical outcomes and reduced costs for ITP patients in the near-term, the long-term consequences of splenectomy remain to be determined. Partial splenectomy has been shown to be effective in the management of anemia in hereditary spherocytosis and elliptocytosis, while preserving vital splenic phagocytic and immune functions. The concept that cell destruction occurs in reticuloendothelial cells has been updated with recognition that the mononuclear phagocyte is neither a reticular nor an endothelial cell. Immune phagocytosis is now understood to be mediated by macrophage IgG Fc and complement receptors. A key factor for devising a strategy for selecting medical or surgical splenectomy, or postponing splenectomy, is an assessment of the relative importance of splenic immune versus phagocytic function in the pathogenesis of ITP. PMID- 10676919 TI - The spleen: anatomy and anatomical function. AB - In this review, our knowledge concerning the structure and function of the spleen is summarized. The unique architecture of the spleen allows for interactions between the circulatory, reticuloendothelial, and immune systems. Based on these interactions in conjunction with its microanatomy, the spleen is able to maintain the integrity of the blood and respond to circulating antigens. However, this can be a double-edged sword in the case of patients suffering from autoimmune diseases such as immune thrombocytopenic purpura since the spleen can be the site of both antibody production and circulating cell destruction. PMID- 10676920 TI - Long-term outcome of splenectomy for idiopathic thrombocytopenic purpura. AB - Idiopathic thrombocytopenic purpura (ITP) is an illness of primary acquired thrombocytopenia occurring in the absence of marrow failure. Splenectomy was first used as a treatment for ITP in 1913. However, with the realization that opsonin (critical for the optimal killing of invasive micro-organisms by white blood cells) is manufactured only in the spleen, spontaneous splenic removal was reevaluated and questioned. Splenectomy has a success rate that remains nearly identical (about 50% to 60%) whether it is performed soon after diagnosis or several months or years later. As yet, there is no consistently effective method to predict an individual ITP patient's response to splenectomy. As the time since splenectomy increases, however, the rate of excellent response decreases. Despite pneumococcal vaccination prior to splenectomy, fatal fulminant sepsis is an omnipresent possibility. Because a number of published studies, including the Johns Hopkins experience, have questioned the long-term outcome of splenectomy, splenectomy should not be the first treatment option for ITP patients. It should be performed only after all other therapeutic modalities have been exhausted, and the patient has a platelet count less than 25,000/microL and is hemorrhaging. Once patients have undergone splenectomy, they are Ineligible for potentially excellent treatment such as anti-D globulin or oral tolerance therapy. PMID- 10676921 TI - The potential for treatment of idiopathic thrombocytopenic purpura with anti-D to prevent splenectomy: a predictive cost analysis. AB - Idiopathic thrombocytopenic purpura (ITP) is an autoimmune disorder affecting both children and adults that can be manifested by severe bleeding episodes. Adult ITP patients have a low rate of spontaneous remission, and symptomatic patients commonly undergo splenectomy; however, maintenance therapy may increase the rate of remission, allowing splenectomy to be avoided. Anti-D is a recently licensed treatment for ITP that has the potential to delay, and possibly avoid, the need for splenectomy. We used preliminary data from an ongoing clinical trial to evaluate the costs involved in using anti-D therapy for 1 year with the intent of avoiding the need for splenectomy. We accounted for different possible outcomes at the completion of the clinical trial. An economic model with a theoretical cohort of 100 patients was developed using the model of an ongoing clinical trial. The average wholesale price was used to determine the cost of an infusion of anti-D based on an average dose ($1,213 per infusion). The cost of splenectomy was determined by a literature review ($16,000). Costs were calculated for all known patient outcomes; where outcomes were unknown and likely to vary, all possible outcomes were accounted for (splenectomy or no splenectomy). In our theoretical cohort, 31 of 100 patients were taken off anti-D and received splenectomy, 32 of 100 were stable after receiving anti-D and would not need splenectomy, and 37 of 100 had Indeterminate outcomes after receiving anti-D. When compared with the cost of the hypothetical scenario of initially giving all 100 patients splenectomy ($1.6 million), a minimum of 47 patients would have to avoid splenectomy to result in a cost savings for our cohort of 100 patients. The group of 47 patients avoiding splenectomy would be composed of the 32 patients comprising the stable group and at least 15 of the 37 patients comprising the group with indeterminate outcomes. If all 37 of the patients in the group with indeterminate outcomes avoid splenectomy, $363,000 and 69 spleens would be saved. Our data suggest that in the phase III trial of maintenance anti D therapy versus immediate splenectomy, anti-D therapy will be a cost-effective option if 47% or more of patients avoid splenectomy. PMID- 10676922 TI - Treatment options for chronic idiopathic (immune) thrombocytopenic purpura. AB - The goal of treatment for idiopathic (immune) thrombocytopenic purpura (ITP) is to prevent serious bleeding. Traditionally, corticosteroids have been used as first-line therapy followed by splenectomy. Experience with splenectomy over 60 years shows that approximately two thirds of patients achieve normal platelet counts during the initial observation, but that thrombocytopenia often recurs with longer follow-up. We know that long-term use of corticosteroids can lead to significant morbidities; there is no consensus regarding the appropriate timing or indications for splenectomy. To address the Issue of appropriate use of splenectomy, we designed a multicenter clinical trial that will randomize patients to either standard care, involving prednisone followed by splenectomy, or to a novel regimen of limited prednisone treatment followed by WinRho SDF (Nabi, Boca Raton, FL) (anti-D) therapy to maintain the platelet count in a safe range for 1 year. Anti-D can be administered easily in an outpatient setting with few side effects and can provide predictable, transient increases in platelet count. The hypothesis is that prolonged maintenance therapy with a nontoxic regimen may increase the percentage of patients who will experience a spontaneous remission from thrombocytopenia, thereby avoiding an invasive and permanent surgical procedure, splenectomy, and its potentially life-threatening sequelae. PMID- 10676923 TI - Treatment options for chronic immune (idiopathic) thrombocytopenia purpura in children. AB - Immune (idiopathic) thrombocytopenic purpura (ITP) in children is usually acute and self-limiting, but may become chronic in 10% to 30% of patients. Salient issues in the treatment of childhood chronic ITP (cITP) include the following: the choice of immunomodulatory agent; the child's desire for unrestricted physical activity; interventions to avoid or defer splenectomy; and, finally, choosing when (and how) to perform splenectomy. Treatment for children with cITP during childhood usually is extrapolated from that for acute ITP. Treatment with pooled intravenous immunoglobulin (IVIg) and anti-D immunoglobulin often gives an acute response followed by a predictable decay of platelet count. Corticosteroids usually lead to a platelet increase; however, the associated adverse effects of chronic usage are generally unsatisfactory for most children and adolescents. With pulsed, high-dose corticosteroids, a durable platelet response is the exception, not the rule. More aggressive immunosuppression is usually reserved for patients who are symptomatic and refractory to the above treatments, Including splenectomy. Although the estimated success rate ranges from 70% to 90%, the long-term outcome of splenectomy in children with cITP in not well described. In addition, the risk of fatal postsplenectomy infections is significant. A familiar initial strategy among pediatric hematologists thus involves deferral of splenectomy with the reasonable possibility of spontaneous recovery. Corticosteroids, anti-D, and IVIg are effective, temporizing medical alternatives to splenectomy in treating cITP in children. Quality-of-life measurements in children with cITP may help to stimulate the development of new approaches. PMID- 10676924 TI - Splenectomy-sparing, long-term maintenance with anti-D for chronic immune (idiopathic) thrombocytopenic purpura: the New York Hospital experience. AB - For nonsplenectomized children and adults with chronic or acute immune (idiopathic) thrombocytopenic purpura (ITP), anti-D has been shown to be a safe and effective treatment, providing hemostatic platelet increases in more than 70% of patients. Children had the best results, but all patient groups responded. In our recently published series, the effect of anti-D therapy lasted for more than 21 days in 50% of the responders and for more than 1 month in 37%. The use of anti-D as maintenance therapy was evaluated in a subset of patients. Of those who responded to the initial anti-D infusion, 79 patients (44 children, 35 adults) received 3 consecutive treatments. There were no significant differences in the responses after each infusion. Fifty-eight patients responded to all 3 treatments; of those, 20 (9 children, 11 adults) continued anti-D therapy, receiving an average of 18 infusions each (range, 10 to 36). The overall response rate was 86%. The ease of administration, duration of effect, and infrequent development of tachyphylaxis make anti-D an attractive alternative for maintenance therapy, particularly for children with ITP who have a high rate of spontaneous remissions. Use of anti-D as a means of spearing the spleen or, at least, postponing splenectomy should now be considered as a clinical option in the management of adult patients with chronic ITP. PMID- 10676926 TI - Seeds of consensus. PMID- 10676925 TI - Use of anti-D in immune thrombocytopenic purpura as a means to prevent splenectomy: case reports from two University Hospital Medical Centers. AB - Presented here are 16 case studies of adults with immune (idiopathic) thrombocytopenic purpura (ITP); 5 were treated at Hackensack University Medical Center (HUMC), Hackensack, NJ, and 11 were treated at the Allegheny University Hospital (AUH), Medical College of Pennsylvania. Four of the 5 patients at HUMC had initial transient responses to intravenous immunoglobulin G (IVIg) therapy and required large doses of corticosteroids to maintain platelet counts over 50,000 microL. One elderly patient with systemic lupus erythematosus (SLE) had been treated unsuccessfully with corticosteroids and immunosuppressants to maintain her platelet count over 50,000 microL. All 5 patients were given 1 or 2 doses of anti-D at 50 microg/kg, leading to complete resolution of ITP. Following anti-D therapy, patients were tapered off corticosteroids and currently remain in complete remission with platelet counts over 100,000/ microL. The mechanism of action of anti-D in ITP remains unclear and requires further study. Treatment of the 11 patients at AUH began with corticosteroids, which resulted in no durable therapeutic response. Anti-D was then given at 50 microg/kg, and this provoked an excellent response with a prompt recovery of platelet levels to 100,000/ microL, after which active treatment was halted. Patients were monitored by direct office visit every 3 months unless a clinical indication required an earlier return. If the patient's platelets dropped below 100,000/ microL, they were first given prednisone. As of the last follow-up, all 11 patients remain stable and no patients have required splenectomy. PMID- 10676927 TI - Livermore plans radical rescue for 'mismanaged' laser facility PMID- 10676928 TI - Japan may allow human embryo stem-cell research. PMID- 10676929 TI - Change of government bodes well for NZ science funding. PMID- 10676930 TI - WWW project aims to address worldwide decline in amphibians. PMID- 10676931 TI - Anderson steps down from Wellcome over Oxford row. PMID- 10676932 TI - IBM joins genomics mapping consortium. PMID- 10676933 TI - German science starts facing up to its historical amnesia. PMID- 10676934 TI - Apathy rewards misconduct--and everybody suffers. PMID- 10676935 TI - End of impact factors? PMID- 10676936 TI - Nucleic acids revelation delayed by a sceptic. PMID- 10676938 TI - Decline of the generalist PMID- 10676937 TI - And should be treated the same as genomics. PMID- 10676939 TI - From Caribbean to Clementine. PMID- 10676940 TI - Fluff balls of fire PMID- 10676941 TI - Survey flights in honeybees. PMID- 10676942 TI - Electrons in the looking glass PMID- 10676943 TI - Gene expression in diagnosis. PMID- 10676944 TI - Plant ecology. Alien invaders. PMID- 10676945 TI - Hydrology. The sting in a fractal tail PMID- 10676946 TI - Huntington's disease. A predictor of pathology. PMID- 10676947 TI - Optical control of reactions PMID- 10676948 TI - Longevity record for deep-sea invertebrate. PMID- 10676949 TI - Mitochondria and the death of oocytes. PMID- 10676950 TI - Demethylation of the zygotic paternal genome. PMID- 10676951 TI - Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. AB - Diffuse large B-cell lymphoma (DLBCL), the most common subtype of non-Hodgkin's lymphoma, is clinically heterogeneous: 40% of patients respond well to current therapy and have prolonged survival, whereas the remainder succumb to the disease. We proposed that this variability in natural history reflects unrecognized molecular heterogeneity in the tumours. Using DNA microarrays, we have conducted a systematic characterization of gene expression in B-cell malignancies. Here we show that there is diversity in gene expression among the tumours of DLBCL patients, apparently reflecting the variation in tumour proliferation rate, host response and differentiation state of the tumour. We identified two molecularly distinct forms of DLBCL which had gene expression patterns indicative of different stages of B-cell differentiation. One type expressed genes characteristic of germinal centre B cells ('germinal centre B like DLBCL'); the second type expressed genes normally induced during in vitro activation of peripheral blood B cells ('activated B-like DLBCL'). Patients with germinal centre B-like DLBCL had a significantly better overall survival than those with activated B-like DLBCL. The molecular classification of tumours on the basis of gene expression can thus identify previously undetected and clinically significant subtypes of cancer. PMID- 10676952 TI - Quantum mirages formed by coherent projection of electronic structure AB - Image projection relies on classical wave mechanics and the use of natural or engineered structures such as lenses or resonant cavities. Well-known examples include the bending of light to create mirages in the atmosphere, and the focusing of sound by whispering galleries. However, the observation of analogous phenomena in condensed matter systems is a more recent development, facilitated by advances in nanofabrication. Here we report the projection of the electronic structure surrounding a magnetic Co atom to a remote location on the surface of a Cu crystal; electron partial waves scattered from the real Co atom are coherently refocused to form a spectral image or 'quantum mirage'. The focusing device is an elliptical quantum corral, assembled on the Cu surface. The corral acts as a quantum mechanical resonator, while the two-dimensional Cu surface-state electrons form the projection medium. When placed on the surface, Co atoms display a distinctive spectroscopic signature, known as the many-particle Kondo resonance, which arises from their magnetic moment. By positioning a Co atom at one focus of the ellipse, we detect a strong Kondo signature not only at the atom, but also at the empty focus. This behaviour contrasts with the usual spatially-decreasing response of an electron gas to a localized perturbation. PMID- 10676953 TI - Experimental test of quantum nonlocality in three-photon Greenberger-Horne Zeilinger entanglement AB - Bell's theorem states that certain statistical correlations predicted by quantum physics for measurements on two-particle systems cannot be understood within a realistic picture based on local properties of each individual particle-even if the two particles are separated by large distances. Einstein, Podolsky and Rosen first recognized the fundamental significance of these quantum correlations (termed 'entanglement' by Schrodinger) and the two-particle quantum predictions have found ever-increasing experimental support. A more striking conflict between quantum mechanical and local realistic predictions (for perfect correlations) has been discovered; but experimental verification has been difficult, as it requires entanglement between at least three particles. Here we report experimental confirmation of this conflict, using our recently developed method to observe three-photon entanglement, or 'Greenberger-Horne-Zeilinger' (GHZ) states. The results of three specific experiments, involving measurements of polarization correlations between three photons, lead to predictions for a fourth experiment; quantum physical predictions are mutually contradictory with expectations based on local realism. We find the results of the fourth experiment to be in agreement with the quantum prediction and in striking conflict with local realism. PMID- 10676954 TI - Ball lightning caused by oxidation of nanoparticle networks from normal lightning strikes on soil AB - Observations of ball lightning have been reported for centuries, but the origin of this phenomenon remains an enigma. The 'average' ball lightning appears as a sphere with a diameter of 300 mm, a lifetime of about 10 s, and a luminosity similar to a 100-W lamp. It floats freely in the air, and ends either in an explosion, or by simply fading from view. It almost invariably occurs during stormy weather. Several energy sources have been proposed to explain the light, but none of these models has succeeded in explaining all of the observed characteristics. Here we report a model that potentially accounts for all of those properties, and which has some experimental support. When normal lightning strikes soil, chemical energy is stored in nanoparticles of Si, SiO or SiC, which are ejected into the air as a filamentary network. As the particles are slowly oxidized in air, the stored energy is released as heat and light. We investigated this basic process by exposing soil samples to a lightning-like discharge, which produced chain aggregates of nanoparticles: these particles oxidize at a rate appropriate for explaining the lifetime of ball lightning. PMID- 10676955 TI - Large-scale complementary integrated circuits based on organic transistors AB - Thin-film transistors based on molecular and polymeric organic materials have been proposed for a number of applications, such as displays and radio-frequency identification tags. The main factors motivating investigations of organic transistors are their lower cost and simpler packaging, relative to conventional inorganic electronics, and their compatibility with flexible substrates. In most digital circuitry, minimal power dissipation and stability of performance against transistor parameter variations are crucial. In silicon-based microelectronics, these are achieved through the use of complementary logic-which incorporates both p- and n-type transistors-and it is therefore reasonable to suppose that adoption of such an approach with organic semiconductors will similarly result in reduced power dissipation, improved noise margins and greater operational stability. Complementary inverters and ring oscillators have already been reported. Here we show that such an approach can realize much larger scales of integration (in the present case, up to 864 transistors per circuit) and operation speeds of approximately 1 kHz in clocked sequential complementary circuits. PMID- 10676956 TI - Fractal stream chemistry and its implications for contaminant transport in catchments AB - The time it takes for rainfall to travel through a catchment and reach the stream is a fundamental hydraulic parameter that controls the retention of soluble contaminants and thus the downstream consequences of pollution episodes. Catchments with short flushing times will deliver brief, intense contaminant pulses to downstream waters, whereas catchments with longer flushing times will deliver less intense but more sustained contaminant fluxes. Here we analyse detailed time series of chloride, a natural tracer, in both rainfall and runoff from headwater catchments at Plynlimon, Wales. We show that, although the chloride concentrations in rainfall have a white noise spectrum, the chloride concentrations in streamflow exhibit fractal 1/f scaling over three orders of magnitude. The fractal fluctuations in tracer concentrations indicate that these catchments do not have characteristic flushing times. Instead, their travel times follow an approximate power-law distribution implying that they will retain a long chemical memory of past inputs. Contaminants will initially be flushed rapidly, but then low-level contamination will be delivered to streams for a surprisingly long time. PMID- 10676957 TI - Rapid diffusive infiltration of sodium into partially molten peridotite AB - Recent seismological, geochemical and experimental observations suggest that, as mantle peridotite melts, the resulting basaltic liquid forms an interconnected network, culminating in the rapid ascent of the basalt relative to the surrounding solid matrix. Mantle melting is therefore a polybaric process, with melts produced over a range of pressures having differing chemical characteristics. Modelling and peridotite-melting experiments designed to simulate polybaric mantle melting generally assume that there is no interaction between melts generated at greater pressures and the overlying solid mantle at lower pressures. Beneath mid-ocean ridges, melts derived from greater depth are probably channelized during ascent, so preventing direct re-equilibration with shallow peridotite, as required by geochemical observations. I show here, however, that sodium in ascending melts will quickly diffuse into the melt formed within nearby peridotite at lower pressures. This process fundamentally changes the manner by which the peridotite melts, and can account for both the creation of silica-rich glass inclusions in mantle xenoliths and the anomalous melting modes recorded by abyssal peridotites. Increased melting of lithosphere and upwelling asthenosphere could result from this process without the need to invoke higher mantle temperatures. PMID- 10676958 TI - Oxygen-isotope evidence for recycled crust in the sources of mid-ocean-ridge basalts AB - Mid-ocean-ridge basalts (MORBs) are the most abundant terrestrial magmas and are believed to form by partial melting of a globally extensive reservoir of ultramafic rocks in the upper mantle. MORBs vary in their abundances of incompatible elements (that is, those that partition into silicate liquids during partial melting) and in the isotopic ratios of several radiogenic isotope systems. These variations define a spectrum between 'depleted' and 'enriched' compositions, characterized by respectively low and high abundances of incompatible elements. Compositional variations in the sources of MORBs could reflect recycling of subducted crustal materials into the source reservoir, or any of a number of processes of intramantle differentiations. Variations in (18)O/(16)O (principally sensitive to the interaction of rocks with the Earth's hydrosphere) offer a test of these alternatives. Here we show that (18)O/(16)O ratios of MORBs are correlated with aspects of their incompatible-element chemistry. These correlations are consistent with control of the oxygen-isotope and incompatible-element geochemistry of MORBs by a component of recycled crust that is variably distributed throughout their upper mantle sources. PMID- 10676959 TI - Quality of the fossil record through time. AB - Does the fossil record present a true picture of the history of life, or should it be viewed with caution? Raup argued that plots of the diversification of life were an illustration of bias: the older the rocks, the less we know. The debate was partially resolved by the observation that different data sets gave similar patterns of rising diversity through time. Here we show that new assessment methods, in which the order of fossils in the rocks (stratigraphy) is compared with the order inherent in evolutionary trees (phylogeny), provide a more convincing analytical tool: stratigraphy and phylogeny offer independent data on history. Assessments of congruence between stratigraphy and phylogeny for a sample of 1,000 published phylogenies show no evidence of diminution of quality backwards in time. Ancient rocks clearly preserve less information, on average, than more recent rocks. However, if scaled to the stratigraphic level of the stage and the taxonomic level of the family, the past 540 million years of the fossil record provide uniformly good documentation of the life of the past. PMID- 10676960 TI - Ontogeny of orientation flight in the honeybee revealed by harmonic radar. AB - Cognitive ethology focuses on the study of animals under natural conditions to reveal ecologically adapted modes of learning. But biologists can more easily study what an animal learns than how it learns. For example, honeybees take repeated 'orientation' flights before becoming foragers at about three weeks of age. These flights are a prerequisite for successful homing. Little is known about these flights because orienting bees rapidly fly out of the range of human observation. Using harmonic radar, we show for the first time a striking ontogeny to honeybee orientation flights. With increased experience, bees hold trip duration constant but fly faster, so later trips cover a larger area than earlier trips. In addition, each flight is typically restricted to a narrow sector around the hive. Orientation flights provide honeybees with repeated opportunities to view the hive and landscape features from different viewpoints, suggesting that bees learn the local landscape in a progressive fashion. We also show that these changes in orientation flight are related to the number of previous flights taken instead of chronological age, suggesting a learning process adapted to changes in weather conditions, flower availability and the needs of bee colonies. PMID- 10676961 TI - Myoglobin-like aerotaxis transducers in Archaea and Bacteria. AB - Haem-containing proteins such as haemoglobin and myoglobin play an essential role in oxygen transport and storage. Comparison of the amino-acid sequences of globins from Bacteria and Eukarya suggests that they share an early common ancestor, even though the proteins perform different functions in these two kingdoms. Until now, no members of the globin family have been found in the third kingdom, Archaea. Recent studies of biological signalling in the Bacteria and Eukarya have revealed a new class of haem-containing proteins that serve as sensors. Until now, no haem-based sensor has been described in the Archaea. Here we report the first myoglobin-like, haem-containing protein in the Archaea, and the first haem-based aerotactic transducer in the Bacteria (termed HemAT-Hs for the archaeon Halobacterium salinarum, and HemAT-Bs for Bacillus subtilis). These proteins exhibit spectral properties similar to those of myoglobin and trigger aerotactic responses. PMID- 10676962 TI - Motor disorder in Huntington's disease begins as a dysfunction in error feedback control. AB - A steady progression of motor dysfunction takes place in Huntington's disease (HD). The origin of this disturbance with relation to the motor control process is not understood. Here we studied reaching movements in asymptomatic HD gene carriers (AGCs) and subjects with manifest HD. We found that movement jerkiness, which characterizes the smoothness and efficiency of motion, was a sensitive indicator of presymptomatic HD progression. A large fraction of AGCs displayed elevated jerk even when more than seven years remained until predicted disease onset. Movement termination was disturbed much more than initiation and was highly variable from trial to trial. Analysis of this variability revealed that the sensitivity of end-movement jerk to subtle, self-generated early-movement errors was greater in HD subjects than in controls. Additionally, we found that HD corrective responses to externally-generated force pulses were greatly disturbed, indicating that HD subjects display aberrant responses to both external and self-generated errors. Because feedback corrections are driven by error and are delayed such that they predominantly affect movement termination, these findings suggest that a dysfunction in error correction characterizes the motor control deficit in early HD. This dysfunction may be observed years before clinical disease onset and grows worse as the disease progresses. PMID- 10676963 TI - A neuronal analogue of state-dependent learning. AB - State-dependent learning is a phenomenon in which the retrieval of newly acquired information is possible only if the subject is in the same sensory context and physiological state as during the encoding phase. In spite of extensive behavioural and pharmacological characterization, no cellular counterpart of this phenomenon has been reported. Here we describe a neuronal analogue of state dependent learning in which cortical neurons show an acetylcholine-dependent expression of an acetylcholine-induced functional plasticity. This was demonstrated on neurons of rat somatosensory 'barrel' cortex, whose tunings to the temporal frequency of whisker deflections were modified by cellular conditioning. Pairing whisker stimulation with acetylcholine applied iontophoretically yielded selective lasting modification of responses, the expression of which depended on the presence of exogenous acetylcholine. Administration of acetylcholine during testing revealed frequency-specific changes in response that were not expressed when tested without acetylcholine or when the muscarinic antagonist, atropine, was applied concomitantly. Our results suggest that both acquisition and recall can be controlled by the cortical release of acetylcholine. PMID- 10676964 TI - Modulation of A-type potassium channels by a family of calcium sensors. AB - In the brain and heart, rapidly inactivating (A-type) voltage-gated potassium (Kv) currents operate at subthreshold membrane potentials to control the excitability of neurons and cardiac myocytes. Although pore-forming alpha subunits of the Kv4, or Shal-related, channel family form A-type currents in heterologous cells, these differ significantly from native A-type currents. Here we describe three Kv channel-interacting proteins (KChIPs) that bind to the cytoplasmic amino termini of Kv4 alpha-subunits. We find that expression of KChIP and Kv4 together reconstitutes several features of native A-type currents by modulating the density, inactivation kinetics and rate of recovery from inactivation of Kv4 channels in heterologous cells. All three KChIPs co-localize and co-immunoprecipitate with brain Kv4 alpha-subunits, and are thus integral components of native Kv4 channel complexes. The KChIPs have four EF-hand-like domains and bind calcium ions. As the activity and density of neuronal A-type currents tightly control responses to excitatory synaptic inputs, these KChIPs may regulate A-type currents, and hence neuronal excitability, in response to changes in intracellular calcium. PMID- 10676965 TI - Slowed recovery of rod photoresponse in mice lacking the GTPase accelerating protein RGS9-1. AB - Timely deactivation of the alpha-subunit of the rod G-protein transducin (Galphat) is essential for the temporal resolution of rod vision. Regulators of G protein signalling (RGS) proteins accelerate hydrolysis of GTP by the alpha subunits of heterotrimeric G proteins in vitro. Several retinal RGS proteins can act in vitro as GTPase accelerating proteins (GAP) for Galphat. Recent reconstitution experiments indicate that one of these, RGS9-1, may account for much of the Galphat GAP activity in rod outer segments (ROS). Here we report that ROS membranes from mice lacking RGS9-1 hydrolyse GTP more slowly than ROS membranes from control mice. The Gbeta5-L protein that forms a complex with RGS9 1 was absent from RGS9-/- retinas, although Gbeta5-L messenger RNA was still present. The flash responses of RGS9-/- rods rose normally, but recovered much more slowly than normal. We conclude that RGS9-1, probably in a complex with Gbeta5-L, is essential for acceleration of hydrolysis of GTP by Galphat and for normal recovery of the photoresponse. PMID- 10676966 TI - Food and metabolic signalling defects in a Caenorhabditis elegans serotonin synthesis mutant. AB - The functions of serotonin have been assigned through serotonin-receptor-specific drugs and mutants; however, because a constellation of receptors remains when a single receptor subtype is inhibited, the coordinate responses to modulation of serotonin levels may be missed. Here we report the analysis of behavioural and neuroendocrine defects caused by a complete lack of serotonin signalling. Analysis of the C. elegans genome sequence showed that there is a single tryptophan hydroxylase gene (tph-1)-the key enzyme for serotonin biosynthesis. Animals bearing a tph-1 deletion mutation do not synthesize serotonin but are fully viable. The tph-1 mutant shows abnormalities in behaviour and metabolism that are normally coupled with the sensation and ingestion of food: rates of feeding and egg laying are decreased; large amounts of fat are stored; reproductive lifespan is increased; and some animals arrest at the metabolically inactive dauer stage. This metabolic dysregulation is, in part, due to downregulation of transforming growth factor-beta and insulin-like neuroendocrine signals. The action of the C. elegans serotonergic system in metabolic control is similar to mammalian serotonergic input to metabolism and obesity. PMID- 10676967 TI - Evidence for stabilizing selection in a eukaryotic enhancer element. AB - Eukaryotic gene expression is mediated by compact cis-regulatory modules, or enhancers, which are bound by specific sets of transcription factors. The combinatorial interaction of these bound transcription factors determines time- and tissue-specific gene activation or repression. The even-skipped stripe 2 element controls the expression of the second transverse stripe of even-skipped messenger RNA in Drosophila melanogaster embryos, and is one of the best characterized eukaryotic enhancers. Although even-skipped stripe 2 expression is strongly conserved in Drosophila, the stripe 2 element itself has undergone considerable evolutionary change in its binding-site sequences and the spacing between them. We have investigated this apparent contradiction, and here we show that two chimaeric enhancers, constructed by swapping the 5' and 3' halves of the native stripe 2 elements of two species, no longer drive expression of a reporter gene in the wildtype pattern. Sequence differences between species have functional consequences, therefore, but they are masked by other co-evolved differences. On the basis of these results, we present a model for the evolution of eukaryotic regulatory sequences. PMID- 10676968 TI - Structure of human guanylate-binding protein 1 representing a unique class of GTP binding proteins. AB - Interferon-gamma is an immunomodulatory substance that induces the expression of many genes to orchestrate a cellular response and establish the antiviral state of the cell. Among the most abundant antiviral proteins induced by interferon gamma are guanylate-binding proteins such as GBP1 and GBP2. These are large GTP binding proteins of relative molecular mass 67,000 with a high-turnover GTPase activity and an antiviral effect. Here we have determined the crystal structure of full-length human GBP1 to 1.8 A resolution. The amino-terminal 278 residues constitute a modified G domain with a number of insertions compared to the canonical Ras structure, and the carboxy-terminal part is an extended helical domain with unique features. From the structure and biochemical experiments reported here, GBP1 appears to belong to the group of large GTP-binding proteins that includes Mx and dynamin, the common property of which is the ability to undergo oligomerization with a high concentration-dependent GTPase activity. PMID- 10676969 TI - Alternative modular polyketide synthase expression controls macrolactone structure. AB - Modular polyketide synthases are giant multifunctional enzymes that catalyse the condensation of small carboxylic acids such as acetate and propionate into structurally diverse polyketides that possess a spectrum of biological activities. In a modular polyketide synthase, an enzymatic domain catalyses a specific reaction, and three to six enzymatic domains involved in a condensation processing cycle are organized into a module. A fundamental aspect of a modular polyketide synthase is that its module arrangement linearly specifies the structure of its polyketide product. Here we report a natural example in which alternative expression of the pikromycin polyketide synthase results in the generation of two macrolactone structures. Expression of the full-length modular polyketide synthase PikAIV in Streptomyces venezuelae generates the 14-membered ring macrolactone narbonolide, whereas expression of the amino-terminal truncated form of PikAIV leads to 'skipping' of the final condensation cycle in polyketide biosynthesis to generate the 12-membered ring macrolactone 10-deoxymethynolide. Our findings provide insight into the structure and function of modular polyketide synthases, as well as a new set of tools to generate structural diversity in polyketide natural products. PMID- 10676970 TI - From the CDC: Syphilis elimination: history in the making--opening remarks. PMID- 10676971 TI - From the CDC: Syphilis elimination: history in the making--closing remarks. PMID- 10676972 TI - Syphilis in Atlanta during an era of declining incidence. AB - BACKGROUND: Syphilis transmission in Atlanta is ongoing despite declining incidence. OBJECTIVES: To identify risk factors and missed opportunities for prevention. STUDY DESIGN: A case-control study design was used. Twenty-five sexually transmitted disease (STD) clinic patients with primary or secondary syphilis by polymerase chain reaction and serology and 49 matched controls were interviewed. RESULTS: Persons with syphilis more frequently had HIV infection (24% versus 2%; P = 0.005), crack-using sex partners (52% versus 18%; odds ratio [OR] = 5.1; 95% CI = 1.7-15.5), and a history of incarceration (80% versus 57%; OR = 3.0; CI = 1.0-9.3). Many cases (48%) and controls (31%) had received drug abuse treatment. Only 40% of previously incarcerated patients and 74% of those with a history of drug treatment reported receiving STD/HIV education in those settings. Among all patients reporting recent HIV education, 41% were told about STD recognition and treatment. Unprotected sex and delay in seeking care were common. CONCLUSION: To prevent syphilis and associated HIV, more extensive STD education is needed in jails and drug-treatment centers. PMID- 10676973 TI - Effect of chlorhexidine on genital microflora, Neisseria gonorrhoeae, and Trichomonas vaginalis in vitro. AB - BACKGROUND: Chlorhexidine is a disinfectant that has been used in skin and mouth washes and as a preservative in some vaginal lubricants. A gel containing 0.25% chlorhexidine gluconate has been found to be effective against Chlamydia trachomatis in vitro and in animal models. Applied vaginally, 5 g of this gel could achieve vaginal fluid concentrations of < or = 1250 microg/ml. GOAL: To test the in vitro activity of chlorhexidine in a gel over a pH range of 4 to 8 in the presence or absence of blood. STUDY DESIGN: Organisms were exposed to chlorhexidine for 30 minutes to 2 hours, and the minimum cidal concentration (MCC) was calculated. RESULTS: The MCC for Neisseria gonorrhoeae was 25 microg/ml at 30 minutes and 12.5 microg/ml at 1 to 2 hours of exposure, whereas the MCC for Trichomonas vaginalis was 1250 microg/ml. Chlorhexidine was more active at pH 8 than pH 4, and less active in the presence of blood. The MCC for Lactobacillus crispatus was 1250 microg/ml at pH 4 and only 125 microg/ml at pH 8. CONCLUSIONS: Based on its in vitro activity, chlorhexidine may be an appropriate topical microbicide for prevention of gonorrhea, but not for prevention of trichomoniasis. This study suggests that the presence of blood and pH affect the activity of chlorhexidine against genital pathogens and commensals. PMID- 10676974 TI - Determinants of low-risk and high-risk cervical human papillomavirus infections in Montreal University students. AB - BACKGROUND: Previous studies have been inconsistent about the degree of sexual transmissibility of cervical human papillomavirus (HPV) infection. The authors hypothesize that risk factors for HPV infection vary according to HPV type. GOAL: To estimate the prevalence of HPV infection in asymptomatic women and to identify risk factors for overall HPV infection and HPV infection by oncogenic and nononcogenic type. STUDY DESIGN: A cross-sectional survey was conducted at the McGill University clinic in Montreal. Cervical specimens were collected from 489 female students presenting at the clinic for a routine Papanicolaou test. Data on potential risk factors was obtained by questionnaire. Human papillomavirus DNA was detected by the polymerase chain reaction using consensus primers (MY09/11) followed by hybridization with generic and type-specific probes using Southern blot and dot blot techniques. RESULTS: The overall HPV prevalence was 21.8%. A low-risk HPV infection was found in 6.2% of the women, 11.8% had a high-risk HPV infection (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58), 7.1% had an unknown HPV type, and 2.7% had a multiple type infection. Two profiles emerged for sexual activity and risk of HPV infection according to oncogenic risk after multivariate analysis. Lifetime frequency of sexual intercourse and lifetime number of oral sex partners was associated with high-oncogenic-risk HPV infections; however, HPV infection with low-oncogenic-risk types was invariant with respect to markers of sexual activity. CONCLUSION: These results suggest that there are differences in epidemiologic correlates of transmission between low-risk and high-oncogenic-risk HPV types based on oncogenicity. This finding has important implications for primary prevention of HPV infection and cervical cancer precursors. PMID- 10676975 TI - Risk factors for HTLV-I and II in individuals attending a clinic for sexually transmitted diseases. AB - BACKGROUND: To date, few studies have provided information on risk factors for human t-lymphotropic viruses (HTLV) types I and II in European countries. In particular, few data are available from published studies conducted in STD centers. GOALS: To identify risk factors for HTLV-I and HTLV-II infection and to better distinguish the epidemiologic patterns of the two viruses in Italy. STUDY DESIGN: A cross-sectional study of individuals at high risk of sexually or parenterally transmitted infections attending a large STD center in an urban setting was conducted. Serologic tests for HTLV-I and II, HIV, hepatitis virus type B (HBV), hepatitis virus type C (HCV), and syphilis were performed. Information regarding at-risk behavior was collected using a specific questionnaire. RESULTS: From January 1994 to June 1996, 1,457 individuals were recruited; of them, 1,016 (69.7%) were males, 1,051 (72.4%) Italians, and 288 (19.8%) non-Europeans. One thousand seventy-five (74.8%) participants were noninjecting-drug-using heterosexuals, 285 (19.6%) were men who have sex with men, and 97 (6.6%) were injecting drug users (IDU). The mean age of the study participants was 33.6 (+/-10.5) years. Nine (0.6%) individuals were positive for HTLV-I antibodies and 9 (0.6%) for HTLV-II antibodies. The prevalence of HTLV-I among IDUs, men who have sex with men, and noninjecting-drug-using heterosexuals, was 2.1% (2/97), 1.4% (4/ 285), and 0.3% (3/1085), respectively. HTLV-II prevalence was 8.2% (8/97) among IDUs and 0.09% (1/1075) among noninjecting-drug using heterosexuals. Among the nine HTLV-II-positive individuals, eight were Italian IDUs and one was a noninjecting-drug-using heterosexual man from India. None of the 285 men who have sex with men had HTLV-II antibodies. HTLV-infected individuals tended to be older than those who were uninfected. HTLV-I-infected individuals were more likely to be non-European and to have antibodies against Treponema pallidum. Injecting drug use tended to be independently associated with HTLV-II infection. CONCLUSIONS: The data suggest a role of sexual behavior in the spread of HTLV-I, which is more likely to be detected in individuals coming from endemic areas. Injecting drug use remains the most important risk factor for HTLV II infection in Italy. PMID- 10676976 TI - Predictors of female condom use among women exchanging street sex in New York City. AB - BACKGROUND: Alternative female-initiated barrier methods, such as the female condom, are needed among women exchanging street sex to enhance their ability to protect themselves from HIV and STD infection. OBJECTIVE: To describe predictors of female condom use among 96 women exchanging sex for money and drugs on the streets of New York City. STUDY DESIGN: A total of 113 sex workers received a baseline interview, a demonstration on proper female condom use, and 10 female condoms. A total of 101 sex workers received a followed-up evaluation at 2 weeks, of which 96 were included in data analysis. Predictors of condom use were analyzed for (1) any type of use; and (2) use with commercial partners. RESULTS: The strongest predictors of female condom use among this sample of sex workers were (1) living with someone with a drug or alcohol problem; (2) having heard of the female condom; and (3) homelessness. Current physical or sexual abuse by a commercial partner and marriage decreased the probability of female condom use. CONCLUSIONS: Female condom distribution encouraged sex workers who may be most vulnerable or who reported characteristics or behaviors associated with the highest sexually transmitted disease and HIV risk to try female condoms with commercial partners. Implications for intervention development include the need to develop innovative programs provided on the street (e.g., through peers) that can access homeless, drug-using sex workers in the most at-risk environments. PMID- 10676977 TI - Evaluating AIDS prevention interventions using behavioral and biological outcome measures. AB - OBJECTIVE: To begin a dialogue on the role of behavioral and biological outcome measures in evaluating the effectiveness of behavior change interventions to reduce the risk of transmitting and acquiring HIV and other sexually transmitted diseases (STDs). METHODS: A selective review of the literature was undertaken to identify issues and problems associated with the use of behavioral and biological outcome measures. In particular, the article considers the validity of self reports and the theoretical relationships between behavioral and biological measures. RESULTS: Available data suggest that when proper care is taken, behavioral self-reports are valid. Similarly, sensitive and specific diagnostic tests are available, particularly for bacterial STDs. However, even when diagnostic tests and behavioral self-reports provide valid data, one should not expect a simple relationship between behavioral and biological measures. CONCLUSION: Both behavioral and biological measures are important outcomes for studying the efficacy and effectiveness of behavior-change interventions. However, one measure cannot substitute for or validate the other, and neither serves as a true surrogate for HIV prevalence or incidence. Therefore, it is important to better understand the relationship among STDs, HIV, and self reported condom use. To do this, it will first be necessary to assess correct as well as consistent condom use. PMID- 10676978 TI - Cumulative experience with Haemophilus ducreyi 35000 in the human model of experimental infection. AB - BACKGROUND AND OBJECTIVES: To study Haemophilus ducreyi pathogenesis, the authors developed an experimental model of infection in human volunteers. The authors analyze their cumulative experience with strain 35000 in the model and calculate the papule and pustule formation rates for estimated delivered doses (EDDs) ranging from 1 cfu to 100 cfu. STUDY DESIGN: Sixty-five volunteers were included in the analysis. A total of 139 sites were available for calculation of the papule formation rate, and 117 sites were available for calculation of the pustule formation rates. RESULTS: The effect of EDDs and probabilities of papule formation and the pustule formation were dose-dependent. Increasing the EDD resulted in a higher probability of papule and pustule formation. CONCLUSION: H ducreyi is highly infectious for humans. Inoculation of an EDD of 1 cfu causes a papule formation rate of 50%. Pustule formation rates are approximately 50% for 27 cfu and 90% for 100 cfu. PMID- 10676979 TI - Resistance of Neisseria gonorrhoeae epidemic strains to antibiotics: report of resistant isolates and surveillance in Zhanjiang, China: 1998 to 1999. AB - BACKGROUND: Antibiotics are widely used to treat gonorrhea. Changes in the susceptibility of Neisseria gonorrhoeae to these agents may influence their use. GOAL: To measure the antibiotic susceptibility of N gonorrhoeae epidemic strains in Zhanjiang (Guangdong) and to evaluate the prevalence of strains with reduced susceptibility. STUDY DESIGN: A total of 98 gonococcal isolates obtained from 1998 through 1999 in Zhanjiang were tested for antibiotic susceptibility based on the systemic identification. The inhibitory zone diameters (mm) and the MICs of penicillin, tetracycline, spectinomycin, ceftriaxone, and ciprofloxacin were determined using disk-diffusion and agar-dilution methods, respectively. The susceptibilities of these isolates were defined using criteria of the National Committee for Clinical Laboratory Standards. RESULTS: The percentages of gonococci-resistant strains to penicillin, tetracycline, spectinomycin, ceftriaxone, and ciprofloxacin were 32.65%, 69.39%, 8.16%, 13.27%, 82.65% by disk diffusion method and 23.91%, 49.46 %, 11.11%, 16.48%, 59.34% by agar-dilution method, respectively. CONCLUSIONS: The resistant strains of contemporary gonococci in Zhanjiang were serious, especially for ciprofloxacin resistance. Continued active surveillance is needed to detect and control the spread of ceftriaxone-resistant and spectinomycin-resistant N gonorrhoeae. PMID- 10676980 TI - Acceptability of formulations and application methods for vaginal microbicides among drug-involved women: results of product trials in three cities. AB - BACKGROUND AND OBJECTIVES: Female-controlled methods of HIV prevention, such as vaginal microbicides, are urgently needed, particularly among drug-involved women. Acceptability research is critical to product development. GOAL: To assess the acceptability of forms and application methods for future microbicides. DESIGN: Eighty-four drug-involved women were introduced in groups to three lubricant products, asked to try each for 3 weeks, and scheduled for individual follow-up interviews. RESULTS: Participants and their partners felt positive about the products, and expressed willingness to use microbicides if they were shown to be effective against HIV. Women agreed on product characteristics that influenced their reactions (e.g. ease of insertion, degree of "messiness"), but often disagreed on whether their reactions to these characteristics were positive or negative. CONCLUSION: Development of acceptable and effective HIV-prevention products depends on understanding the interaction between characteristics of the products and the characteristics and perceptions of women. Levels of sexual risk and acceptability factors based on drug-use patterns, race and ethnicity, culture, age, and types and attitudes of male partners suggest that a "one size fits all" approach will not win broad acceptance among drug-involved women. PMID- 10676981 TI - Direct injection of physiological fluids in micellar liquid chromatography. AB - Micellar liquid chromatography (MLC), which uses mobile phases of surfactants above the critical micellar concentration, provides a solution to the direct injection of physiological samples by solubilizing the protein components, and coating the analytical column with surfactant monomers to avoid clogging. A review showing the advantages and limitations of this technique over other chromatographic techniques used in drug analysis, working protocols, and examples of application is presented. The possibility of direct sample introduction simplifies and greatly expedites the treatments with reduced cost, improving the accuracy of the procedures. Surfactant monomers and micelles appear to displace drugs bound to proteins, releasing them for partitioning to the stationary phase. The versatility of MLC encompasses the wide range of drug classes normally monitored, such as analgesics, anticancer drugs, antidepressants, bacteriostats, beta-blockers, bronchodilators, catecholamines, diuretics and steroids, among others. Analytical procedures have been developed in urine, plasma, serum and cow milk samples. Most of them utilize sodium dodecyl sulphate as surfactant and a C18 column. UV detection is usual, but enhanced detection has been reported by measuring the absorbance in the visible region of drug derivatives formed precolumn, and with a variety of other techniques, such as fluorimetry, amperometry, inductively coupled plasma-mass spectrometry and immunoassay. Column switching with on-line surfactant-mediated sample clean-up is shown as an attractive enrichment technique, which expands the practical use of MLC beyond the singular dimensional chromatographic process. PMID- 10676982 TI - Quantitation of cerivastatin and its seven acid and lactone biotransformation products in human serum by liquid chromatography-electrospray tandem mass spectrometry. AB - A method for the simultaneous quantitation of cerivastatin (acid) and its biotransformation products, cerivastatin lactone, M-1 (acid), M-1 lactone, M-23 (acid), M-23 lactone, M-24 (acid) and M-24 lactone, in human serum by high performance liquid chromatography (LC) with positive ion electrospray tandem mass spectrometry (MS-MS) was developed and validated. The method involves extraction of cerivastatin and its biotransformation products from acidified human serum (0.5 ml) using methyl tert.-butyl ether. The standard curve ranges in human serum were from 0.0100 to 10.0 ng/ml for cerivastatin and cerivastatin lactone, 0.0500 to 10.0 ng/ml for M-1 (acid) and M-1 lactone, 0.100 to 10.0 ng/ml for M-23 (acid) and M-23 lactone, and 0.500 to 10.0 ng/ml for M-24 (acid) and M-24 lactone. The lactone compounds in human serum at room temperature underwent considerable conversion to the corresponding acid compounds after only 4 h. Lowering the serum pH with a pH 5.0 buffer stabilized the lactone compounds for up to 24 h at room temperature. The degree of lactonization of the acid compounds was < or = 3.5% and the degree of hydrolysis of the lactone compounds was < or = 6.0% during the entire assay procedure. All the eight analytes eluted within 2.0 min and the total run time was only 3.5 min. PMID- 10676983 TI - Analysis of melphalan adducts of 2'-deoxynucleotides in calf thymus DNA hydrolysates by capillary high-performance liquid chromatography-electrospray tandem mass spectrometry. AB - Melphalan is a bifunctional alkylating agent that covalently binds with intracellular nucleophilic sites. A methodology using electrospray mass spectrometry was developed to detect and identify DNA adducts. Alkylation sites within a particular nucleotide were examined using electrospray tandem mass spectrometry hyphenated to capillary liquid chromatography in combination with a column switching system. In the reaction mixtures resulting from the interaction of 2'-deoxynucleotides and melphalan several base-aklylated adducts were found. In the case of 2'-deoxyadenosine monophosphate, thymidine monophosphate and 2' deoxyguanosine phosphate alkylation was observed in the mononucleotide reaction mixtures but not in the DNA-hydrolysates. Calf thymus DNA was reacted in vitro with melphalan. The DNA pellet was isolated and enzymatically hydrolyzed with the aid of Nuclease P1. In this hydrolysate both mono-alkylated 2'-deoxynucleotides and dinucleotides were found. The most important adduct found was identified as the N-7 alklylated dGMP adduct. The alkylated dinucleotides were identified as a pdApdT/melphalan and pdGpdC/melphalan the latter being the most important. PMID- 10676984 TI - High-performance liquid chromatographic determination of [11C]1-(2-chlorophenyl) N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide in mouse plasma and tissue and in human plasma. AB - The high-performance liquid chromatographic determination of 1-(2-chlorophenyl)-N methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide ([11C]PK 11195) is described. The method was successfully applied for plasma and tissue analysis after i.v. injection of [11C]PK 11195 in mice and for plasma analysis after administration of [11C]PK 11195 to humans. Separation is effected on a RP-C18 column, using a mixture of acetonitrile-water-triethylamine (65:35:0.5, v/v). Quantitative measurements of radioactivity are performed on a one-channel gamma ray spectrometer equipped with a 2 x 2 in. NaI(Tl) detector. For humans rapid metabolisation of [11C]PK 11195 was observed. At 5, 20 and 35 min post injection 5%, 22% and 32%, respectively, of the plasma activity consisted of at least two more polar metabolites. Despite the extensive metabolisation rate in mice (up to 42% at 10 min post injection of [11C]PK 11195), no 11C-labelled metabolites could be detected in the extracts of brain and heart. PMID- 10676985 TI - Quantitative determination of a potent lipopolysaccharide antagonist, E5564, in rat and dog plasma by high-performance liquid chromatography with fluorescence detection. AB - The assay method was established for the quantification of a potent lipopolysaccharide (LPS) antagonist, E5564, in rat and dog plasma using HPLC. E5564 and the I.S. (an analogue of E5564) were extracted and derivatized with 9 Anthryldiazomethane (ADAM reagent) to be given fluorescence. LC-MS analysis indicated that single molecule of E5564 was coupled with two molecules of ADAM reagent at one on each of the phosphorus groups. After solid-phase extraction, ADAM derivatives of E5564 and the I.S. were separated on an ODS column using methanol/ethanol containing sodium acetate as a mobile phase at 1.2 ml/min (gradient elution), and detected by a fluorescence detector (excitation: 254 nm, emission: 415 nm). The intra-day and inter-day precision were less than 14.4%, and accuracy were within +/-13.0% in the concentration range of 30 to 20,000 ng/ml plasma in both species. E5564 was stable for at least 13 days in rat and dog plasma at -20 degrees C, and the processed sample was stable for up to 14 days at 4 degrees C. This validated method was successfully applied to the evaluation of the pharmacokinetics of E5564 in rats and dogs after single bolus intravenous doses. PMID- 10676986 TI - Separation of human vitamin K-dependent coagulation proteins using hydrophobic interaction chromatography. AB - A rapid and simple method was developed to separate human vitamin K-dependent plasma proteins from each other, yielding virtually homogeneous pools. The purification technique is based on the single use of hydrophobic interaction chromatography, starting from prothrombin concentrate (PC or DEFIX, also termed factor IX concentrate) as initial material. Phenyl-sepharose HP demonstrated optimal separation by comparing several hydrophobic resins as well as resins used in standard procedures like immobilised heparin and Cibacron blue. Under ideal conditions, factor X could be separated in a single step as well as prothrombin. Factor IX co-eluted with other minor proteins. Focus was given only on these three proteins due to their relative abundance. Complete separation of all proteins present in the starting material was achieved by MonoQ anion-exchange chromatography following the phenyl-sepharose run. The resulting purified material could be demonstrated to be of equal or higher purity than using described methods. This strategy employing hydrophobic interaction chromatography for blood macromolecules could be of immense value for purifying the human vitamin K-dependent proteins and represents a considerable simplification over other purification schemes. It not only involves minimal sample handling but also can be readily up-scaled and is a cost-efficient alternative. PMID- 10676987 TI - Determination of tramadol by capillary gas chromatography with flame ionization detection. Application to human and rabbit pharmacokinetic studies. AB - A rapid, sensitive, precise and accurate capillary gas chromatographic assay with flame ionization detection was developed for the determination of tramadol in human, rabbit, pig and dog plasma. It is comprised of only a one-step extraction procedure with dichloromethane at pH 11.15 and gas chromatography on a capillary column. The recoveries of tramadol and meperidine (internal standard) were greater than 88%. Calibration graphs were linear over the concentration range 12.5-10,000 ng/ml with a coefficient of variation, both within-day and between day, of less than 10% at any level. The limit of detection was 8 ng/ml of plasma based on signal-to-noise ratio of 3. Six other clinically used analgesics were investigated to check for potential interferences and their analytical conditions. The specificity of this assay was checked with two major metabolites of tramadol (M1: O-demethyltramadol; M2: N-demethyltramadol). Tramadol in plasma did not decompose significantly at -20 degrees C for 56 days. Pharmacokinetic application with intravenous tramadol in humans and rabbits revealed that tramadol followed a two-compartment open model with one distribution phase and one elimination phase. The distribution and elimination half-lives in humans were 1.02 and 141.9 min. The distribution and elimination half-lives in rabbits were 7.31 and 63.2 min, respectively. PMID- 10676988 TI - Simple and sensitive determination of 5-fluorouracil in plasma by high performance liquid chromatography. Application to clinical pharmacokinetic studies. AB - 5-Fluorouracil (5-FU) is an antineoplastic agent widely employed in the treatment of many types of cancer. Recent studies have proved the need for individual adjustment of 5-FU dosage based on pharmacokinetics. A simple and sensitive high performance liquid chromatographic method for the determination of 5-FU in plasma and their preliminary clinical pharmacokinetics is described. After sample acidification with 20 microl of orthophosphoric acid (5%), the drug is extracted from plasma using n-propanol-diethyl ether (16:84). The organic layer is evaporated to dryness, the residue dissolved in 100 microl of mobile phase and 20 microl of this mixture is injected into a LiChrospher 100RP-18 (5 microm, 250 x 4.0 mm) analytical column. Mobile phase consisted of potassium dihydrogenphosphate (0.05 M, adjusted to pH 3). The limit of quantitation was 2 ng/ml. The method showed good precision: the within-day relative standard deviation (RSD) for 5-FU (10-20,000 ng/ml) was 3.75% (2.57-5.93); the between-day RSD for 5-FU, in the previously described range, was 5.74% (4.35-7.20). The method presented here is accurate, precise and sensitive and it has been successfully applied for 5-FU pharmacokinetic investigation and therapeutic drug monitoring. PMID- 10676989 TI - Validation of urine drug-of-abuse testing methods for ketobemidone using thin layer chromatography and liquid chromatography-electrospray mass spectrometry. AB - High-performance thin-layer chromatography (TLC) with visual detection (post chromatographic derivatization) was used in screening for the drug ketobemidone in human urine samples. High-performance liquid chromatography with electrospray mass spectrometry (LC-ESI-MS) was used for final confirmation of the result. The clean-up was performed by mixed-mode solid-phase extraction, and nalorphine was used as internal standard. A screening cut-off for TLC was established at 0.2 microg/ml. The mean recovery for LC-MS was 91% (n=60) with coefficients of variation (C.V.) in the range of 7 to 16%. Qualifying fragment ions of ketobemidone (m/z 190, 201 and 230) were generated by up front collision-induced dissociation (CID) on a single quadrupole instrument. Relative ion intensities were within +/- 15% deviation compared with standards in the same batch. The limit of detection for LC-MS was 0.025 microg/ml. Positive clinical samples from drug abusers (n=10) had concentrations in the range 0.07 to 3.2 microg/ml, which could be determined by LC-MS without matrix interference. During screening of unknown clinical samples (n=27) the results from TLC was in agreement with LC-MS data. After acid hydrolysis of conjugates in clinical samples the analyte response of ketobemidone and norketobemidone was increased by a factor of approximately two and twelve, respectively. A qualitative GC-MS technique was demonstrated for the detection of the spasmolyticum A29 (N,N-dimethyl-4,4 diphenyl-3-buten-2-amine), which can be found in a preparation combined with ketobemidone (Ketogan). PMID- 10676990 TI - Improved and simplified liquid chromatographic assay for adefovir, a novel antiviral drug, in human plasma using derivatization with chloroacetaldehyde. AB - A rapid and simplified chromatographic assay is reported for the quantification of adefovir (PMEA) utilizing derivatization with chloroacetaldehyde. Adefovir is isolated from plasma using protein precipitation with trichloroacetic acid; next, the fluorescent 1,N6-etheno derivative is directly formed at 98 degrees C in the buffered extract with chloroacetaldehyde. This derivative is analyzed using isocratic ion-pair liquid chromatography and fluorescence detection at 254 nm for excitation and 425 nm for emission. In the evaluated concentration range (10-1000 ng/ml) precisions < or = 5% and accuracies between 95 and 117% were found, using a 0.2-ml volume of plasma. The lower limit of quantification is 10 ng/ml with a intra-assay precision of 16%. The currently reported bioanalytical method is 20 25-fold more sensitive than previously published assays. PMID- 10676991 TI - Quantitative determination of abacavir (1592U89), a novel nucleoside reverse transcriptase inhibitor, in human plasma using isocratic reversed-phase high performance liquid chromatography with ultraviolet detection. AB - Abacavir is a novel nucleoside reverse transcriptase inhibitor for the treatment of HIV-1 infection. A simple and rapid high-performance liquid chromatographic method for the quantification of abacavir in human plasma suitable for pharmacokinetic research purposes is described. Sample pretreatment consists of protein precipitation with perchloric acid. The supernatant is injected directly into the chromatographic system after centrifugation. The drug is separated from endogenous compounds by isocratic reversed-phase high-performance liquid chromatography with ultraviolet detection at 285 nm. The method has been validated over the range of 20-2000 ng/ml using a volume of 300 microl of plasma. The assay is linear over this concentration range as indicated by the F-test for lack-of-fit. Within- and between-day precisions are less than 7.5% for all quality control samples. The lower limit of quantitation is 20 ng/ml and the recovery of abacavir is 88.1% (+/-1.3%). Frequently coadministered drugs did not interfere with the described methodology. Abacavir is stable in human plasma under various relevant storage conditions, for example when stored for 51 days at -20 degrees C. This validated assay is suited for use in pharmacokinetic studies with abacavir in human plasma and can readily be implemented in the setting of a hospital laboratory for the monitoring of abacavir concentrations. PMID- 10676992 TI - Determination of unbound cefmetazole in rat blood by on-line microdialysis and microbore liquid chromatography: a pharmacokinetic study. AB - A specific and sensitive microbore liquid chromatographic method for the determination of unbound cefmetazole in rat blood was developed. A microdialysis probe was inserted into the jugular vein/right atrium of a Sprague-Dawley rat. Cefmetazole (10 mg/kg, i.v.) was then administered via the femoral vein. Dialysates were automatically injected into a liquid chromatographic system via an on-line injector. Isocratic elution of cefmetazole was achieved by LC-UV within 10 min. Intra- and inter-assay accuracy and precision of the assay were < or = 10%. The detection limit of cefmetazole was 20 ng/ml. Pharmacokinetic analysis of results indicated that unbound cefmetazole levels in rats best fit a biexponential decay model. PMID- 10676993 TI - High-performance liquid chromatographic assay for the determination of the novel taxane derivative IDN5109 in mouse plasma. AB - An HPLC assay was developed to determine the paclitaxel analogue 13-(N-tert. butoxycarbonyl-beta-isobutylisoserinyl)-14-hydroxyb accatin-1,14-carbonate (IDN5109) and its epimer in mouse plasma. The method involves solid-phase extraction on cyano cartridges (recovery >75%), HPLC separation on symmetry shield column, a mobile phase of NaH2PO4 (10 mM) pH 5.2, acetonitrile (47:53) and detection at 227 nm. Retention times of IDN5109, its epiform and internal standard were 15, 24 and 25.5 min, respectively. The assay was linear from 0.10 to 10 microg/ml (r2 = 0.999), with a C.V. <5% and accuracy in the range of 95 107%. LOQ was 50 ng/ml for both compounds. Using this method IDN5109 pharmacokinetic was determined in mice. PMID- 10676994 TI - Radiochemical high-performance liquid chromatographic assay for the determination of catechol O-methyltransferase activity towards various substrates. AB - A new chromatographic catechol O-methyltransferase (COMT) assay based on S adenosyl-L-[methyl-14C]methionine and on-line radioactivity detection was developed. With minor modifications in the mobile phase composition the methylation velocities for 30 structurally diverse compounds including simple catechols, neurotransmitters, catecholestrogens and catecholic drugs could be measured using human and rat recombinant soluble COMT. The enzymes showed very similar substrate selectivities. The radiochemical method was validated using 3,4 dihydroxybenzoic acid as a model substrate and it was shown that accurate and reproducible methylation velocity values could be achieved for both of the catecholic hydroxyls. The method proved to be suited for determining the enzyme kinetic parameters and can probably be further used for gathering enzyme kinetic data on differentially substituted catechols in order to construct proper structure-activity relationships for COMT. PMID- 10676995 TI - Aggregation of recombinant hepatitis B surface antigen induced in vitro by oxidative stress. AB - In order to examine whether oxygen radicals could be responsible for aggregation of recombinant hepatitis B surface antigen (HBsAg) during its assembly in yeast, purified HBsAg was oxidized with ammonium peroxodisulphate (AP) and analyzed by non-denaturing and denaturing size exclusion chromatography, immunoassay and immunoelectron microscopy. As a result, peroxodisulphate radicals induced a reproducible aggregation of HBsAg. At 44 mM AP, the aggregation process took a few hours and the resulting structures were large, branched and non-antigenic. During more gentle oxidation with 9 mM AP and 20-80 microM Cu2+, a continuous structural modification to HBsAg delaying for tens of hours preceded the aggregation event. During this pre-aggregation period, peroxidation of HBsAg lipids and covalent cross-linking of S protein chains occurred that led a complete loss of antigenicity of oxidized particles. In contrast, yeast-derived HBsAg aggregate is decomposed to S monomers under reducing conditions and recognized by anti-HBsAg polyclonal and monoclonal antibodies, suggesting that is has been assembled in vivo from antigenic and reversibly cross-linked particles. Based on these observations, we conclude that oxidation, at least with respect to the specific molecular sites oxidized by AP, is not a primary event in HBsAg aggregate formation in vivo. Since oxidized HBsAg was shown to be irreversibly cross-linked and non-antigenic, there are no suitable techniques for detection HBsAg oxidation in biological samples. Hence, at present, the magnitude of the in vivo oxidative damage to HBsAg cannot be evaluated and thus, whether the plasma derived HBsAg undergoes radical-induced oxidation in the course of viral hepatitis remains to be established. If this occurs, this process is expected to contribute to low HBsAg levels in chronic hepatitis B carriers, failure of the currently available immunoassays to identify HBsAg-positive blood donors and inconsistency in the results provided by HBsAg- and anti-HBsAg-based tests in several recent reports. PMID- 10676996 TI - Determination of flunarizine in rat brain by liquid chromatography-electrospray mass spectrometry. AB - A rapid liquid chromatography-electrospray mass spectrometry (LC-ES-MS) assay for the determination of flunarizine (FZ) in rat brain has been developed. A C18 column and an isocratic elution were employed for the separation. Using post column split, 64% of the eluent was introduced into the ES-MS system for detection. The [M+H]+ (m/z 406) and a fragmented ion (m/z 203) were detected using selected ion monitoring. The linear range of this assay was good, ranging from 0.05 to 5 microM (r2=0.99). The intra- and inter-day precisions showed relative standard deviations ranging from 1.4% to 2.0% and 1.3% to 2.9%, respectively. The application of this newly developed method was demonstrated by examining the pharmacokinetics of FZ in rat brain. PMID- 10676997 TI - Sensitive determination of irinotecan (CPT-11) and its active metabolite SN-38 in human serum using liquid chromatography-electrospray mass spectrometry. AB - A couple of sensitive and accurate liquid chromatography-electrospray mass spectrometry (LC- S-MS) methods for the determination of the total forms of irinotecan and its active metabolite SN-38 in human serum, using the same chromatographic and detection conditions, is presented. Both used camptothecin as internal standard (I.S.). The sample pretreatment for irinotecan involved a simple protein precipitation with acetonitrile, whereas a liquid-liquid extraction was necessary for SN-38. A Symmetry C18, 3.5 microm (150 x 1 mm I.D.) reversed-phase column was used for the chromatographic separation, together with a gradient elution of acetonitrile in 5 mM ammonium formate buffer (pH 3) as mobile phase. After ionisation in the pneumatically-assisted electrospray source and in-source collision induced dissociation, acquisition was performed in the selected ion monitoring mode. Recoveries were 69 and 47% on average, detection limits 2.5 and 0.25 ng/ml and quantitation limits 10 and 0.5 ng/ml for irinotecan and SN-38, respectively. Reproducibility was good and the method was linear from limits of quantitation up to 10,000 ng/ml for irinotecan, and up to 100 ng/ml for SN-38. This sensitive and highly specific method is suitable both for pharmacokinetic studies and routine therapeutic drug monitoring. PMID- 10676998 TI - Identification of metallothionein isoforms on capillary zone electrophoresis by adding anti-metallothionein antibody. AB - The aim of this study was to identify metallothionein (MT) isoforms in mouse liver by using capillary zone electrophoresis (CZE). Purified MT-1 and MT-2 isoforms were completely separated by CZE using a polyacrylamide-coated tube at physiologic pH. There were two peaks in the cytosol fraction prepared from zinc injected mouse liver, in which the migration times corresponded with those of purified MT-1 and MT-2 isoforms. When anti-MT monoclonal antibody was added with the purified MT-1 or MT-2 solution, the peaks decreased. Furthermore, the two peaks in the cytosol prepared from Zn-injected mouse liver decreased in a time dependent manner from the electropherogram after the addition of the antibody. Therefore, those peaks were identified as MT-1 and MT-2 isoforms, respectively. In conclusion, the addition of anti-MT monoclonal antibody to the cytosol fraction of tissues is an effective method for identification of MT isoforms after separation using CZE. PMID- 10676999 TI - Highly sensitive high-performance liquid chromatographic assay for methotrexate in the presence and absence of anti-methotrexate antibody fragments in rat and mouse plasma. AB - Recently, Balthasar and Fung have proposed that anti-methotrexate antibody fragments may be employed to enhance the selectivity of intraperitoneal methotrexate (MTX) therapy. This current work presents a sensitive high performance liquid chromatographic method for measuring plasma concentrations of total (i.e., bound and unbound) MTX and free (unbound) MTX in rat and mouse plasma, in the presence or absence of therapeutic anti-MTX antibody fragments. The assay involves pre-column derivatization of MTX by sodium hydrosulfite to 2,4 diamino-6-methylpteridine. The limit of quantitation for MTX by this assay was 1.25 ng in rat plasma, mouse plasma and mouse plasma ultrafiltrate, which corresponds to a concentration of 25 ng/ml for a 50 microl sample. The limit of quantitation was found to be 2.5 ng in rat plasma ultrafiltrate (i.e., 50 ng/ml in 50 microl rat plasma ultrafiltrate). The method was shown to be quite accurate, as the mean assayed concentration of quality control samples was within 10% of theoretical values. We have applied the method to the investigation of MTX pharmacokinetics in mice and rats, following the administration of MTX alone or following simultaneous administration of MTX and anti-MTX Fab fragments. The method has been shown to be suitable for the assay of total and free methotrexate in the plasma of these species and will enable the testing of pharmacokinetic hypotheses regarding the influence of anti-MTX Fab fragments on the disposition of MTX. PMID- 10677000 TI - Stereoselective measurement of E- and Z-doxepin and its N-desmethyl and hydroxylated metabolites by gas chromatography-mass spectrometry. AB - A stereoselective method of analysis of the antidepressant drug doxepin (DOX, an 85:15% mixture of E-Z stereoisomers), its principal metabolites E- and Z-N desmethyldoxepin (desDOX) and ring-hydroxylated metabolites in microsomal incubation mixtures is described. DOX and its metabolites were extracted from alkalinised incubation mixtures by either: 9:1 hexane-propan-2-ol (method 1) or 1:1 hexane-dichloromethane (method 2), derivatised with trifluoroacetic anhydride and analysed by GC-MS with selected ion monitoring. Both methods were suitable for the analysis of individual desDOX isomers as indicated by correlation coefficients of > or = 0.999 for calibration curves constructed between 50 and 2500 nM, and good within-day precision at 125 nM (C.V. < or = 14%) and 1000 nM (C.V. < or = 8%). Method 1, however, was unsuitable for the analysis of ring hydroxylated metabolites of DOX, whereas the hydroxylated metabolites of E-DOX and E-desDOX (generated in situ) were extracted by method 2 with a C.V. of ca. 13%. This is the first assay method that permits the simultaneous measurement of desDOX and hydroxylated metabolites of DOX in microsomal mixtures. PMID- 10677001 TI - Quantitative analysis of clenbuterol in meat products using liquid chromatography electrospray ionisation tandem mass spectrometry. AB - A method is presented that allows quantitation of clenbuterol in meat and liver products at the ng/kg level by liquid chromatography-electrospray ionisation tandem mass spectrometry (LC-ESIMS-MS) using a stable isotopically labeled internal standard. The practical procedure involves acid extraction followed by two solid-phase clean-up steps with C18 and strong cation-exchange (SCX) resins. The typical recovery of the analyte spiked at 0.4 microg/kg in meat and liver samples was at 63+/-7%. Mass spectral acquisition was done in multiple reaction monitoring (MRM) to provide a high degree of sensitivity, achieving a limit of detection and quantitation at 10 and 15 ng/kg, respectively. Two precursor ions at m/z 277 and 279, corresponding to the characteristic isotopic cluster of the two chlorine atoms of clenbuterol, were monitored by LC-ESIMS-MS to provide unambiguous identity of the analyte. Samples of meat and liver of various origins with either incurred residues or spiked with known amounts of clenbuterol were used to validate the method. PMID- 10677002 TI - High-performance thin-layer chromatography method for inositol phosphate analysis. AB - A simple and inexpensive high-performance thin-layer chromatography (HPTLC) method for the analysis of inositol mono- to hexakisphosphates on cellulose precoated plates is described. Plates were developed in 1-propanol-25% ammonia solution-water (5:4:1) and substance quantities as low as 100-200 pmol were detected by molybdate staining. Chromatographic mobilities of nucleotides and phosphorylated carbohydrates were also characterized. Charcoal treatment was employed to separate nucleotides from inositol phosphates with similar R(F) values prior to HPTLC analysis. Practical application of the HPTLC system is demonstrated by analysis of grain extracts from wild type and low-phytate mutant barley as well as phytate degradation products resulting from barley phytase activity. PMID- 10677003 TI - Determination of cetirizine in human plasma by high-performance liquid chromatography. AB - A high-performance liquid chromatographic method for the quantitation of cetirizine in human plasma is presented. The method is based on liquid-liquid extraction with dichloromethane and reversed-phase chromatography with spectrophotometric detection at 232 nm. Gradient elution was used to remove late eluting peaks. Diazepam was used as the internal standard. The limit of quantitation was 10 ng/ml using 0.5 ml of plasma. Within-day and between-day precision expressed by relative standard deviation was less than 10% and inaccuracy did not exceed 8%. The assay was applied to the analysis of samples from a pharmacokinetic study. PMID- 10677004 TI - 3-Hydroxyanthranilic acid-derived compounds formed through electrochemical oxidation. AB - 3-Hydroxyanthranilic acid (3-HAA)-derived oxidation products were analyzed using high-performance liquid chromatography with an electrochemical reactor and diode array detection and high-performance liquid chromatography with an electrochemical reactor and UV detection coupled with mass spectrometry. In addition to 3-HAA dimers such as cinnabarinic acid (CA), 6-amino-3-[(2-carboxy-6 hydroxyphenyl)amino]-2,5-dioxo-1,3-cyclohexadien e-1-carboxylic acid and 4,7 diamino-8-hydroxy-6H-dibenzo[a,d]pyran-6-one-3-carboxylic acid, a 3-HAA trimer and a 3-HAA tetramer were also detected and identified based on their electrospray ionization mass spectra and their UV-visible spectra. These five oxidation products were also detected on the elution profiles of high-performance liquid chromatography-diode array detection analyses for the reaction mixtures of the auto-oxidation of 3-HAA, of 3-HAA with potassium ferricyanide, of 3-HAA with horseradish peroxidase and hydrogen peroxide, and of 3-HAA with superoxide dismutase (SOD). 4,7-Diamino-8-hydroxy-6H-dibenzo[a,d]pyran-6-one-3-carboxylic acid was predominant in the auto-oxidation, in the reaction of 3-HAA with horseradish peroxidase and hydrogen peroxide, and in the electrochemical oxidation of 3-HAA at an applied potential of 0.0 V. On the other hand, CA, the 3 HAA trimer and the 3-HAA tetramer were predominant in the reaction of 3-HAA with K3[Fe(CN)6] and in the electrochemical oxidation of 3-HAA at an applied potential of 1.0 V. PMID- 10677005 TI - Capillary solid-phase extraction-tandem mass spectrometry for fast quantification of free concentrations of tolterodine and two metabolites in ultrafiltered plasma samples. AB - A capillary solid-phase extraction (SPE) system has been coupled directly to electrospray tandem mass spectrometry for quantification of free tolterodine and metabolite concentrations in plasma. The unbound fraction of these compounds was obtained by ultrafiltration of plasma. The ultrafiltrate was directly injected onto the SPE capillary (4 mm x 200 microm, 5 microm C18). After desalting and clean-up of the sample, the analytes were eluted in backflush mode with methanol 1 mM triethylamine (70:30, v/v), providing considerable solute focusing. Elution from the SPE capillary was improved by inserting a short trapping capillary between the SPE capillary and the MS interface, by which analyte focusing was increased. The unresolved compounds eluted simultaneously with the remaining matrix compounds and were detected in a multiple-reaction monitoring (MRM) mode. No interference of the sample matrix on detection was observed, allowing aqueous standards to be used for calibration. Linear calibration curves were obtained between 0.05 and 1000 ng/ml (corresponding to 150 pM-3 microM) and the limit of detection was 50 pg/ml injecting 10 microl. Equilibration of the SPE capillary, sample loading, elution and detection took less then 6 min per sample. PMID- 10677006 TI - Sensitive method for the determination of bisphenol-A in serum using two systems of high-performance liquid chromatography. AB - The aim of this study was to establish an easy and accurate method for the determination of bisphenol-A (BPA) in the body liquid such as serum and urine. Two high-performance liquid chromatography (HPLC) systems, HPLC with electrochemical detector (ED), and HPLC with mass spectrometry (MS) using electrospray ionization (ESI) interface were used for the assay in the serum samples prepared with solid-phase extraction method. Water or EtOH at a concentration below 50% was suitable for the extraction of BPA from serum. The limit of detection of BPA was 0.2 ng ml(-1) for the HPLC-ED method and 0.1 ng ml( 1) for HPLC-MS. There was a good correlation between the data obtained by the two HPLC systems. BPA concentrations in healthy human serum were low (0-1.6 ng ml( 1)). From various commercial fetal bovine serum and sheep plasma, however, significant amounts of BPA were detected. Since no BPA was detected from sheep plasma immediately after collection, the high amounts of BPA were considered to be caused by the handling of blood during the preparation of the products after blood collection. In vitro study showed that the amount of BPA leached from polycarbonate tube into sheep plasma were 40 times larger than those into water and the leached amount of BPA depended on the temperature (37 degrees C>20 degrees C>5 degrees C). PMID- 10677007 TI - Direct detection of endogenous histamine in rat peritoneal mast cells by in capillary derivatization high-performance capillary electrophoresis. AB - A simple method for the detection of endogenous histamine in rat peritoneal mast cells was evaluated using on-line mode in-capillary derivatization high performance capillary electrophoretic (ICD-HPCE) techniques, which were previously developed by our group [S. Oguri et al., J. Chromatogr. A, 787 (1997) 253-260]. The method involves a suspension of peritoneal mast cells (1 x 10(6) cells/ml of saline) collected from a male Wistar rat (eight weeks of age), which are directly introduced into the capillary tube from the anodic end by hydrostatic injection (at 25 cm height, for 2-20 s). When a high-voltage potential (25 kV) is applied to the capillary, which is already filled with the run buffer containing both a lysing reagent (SDS, sodium dodecyl sulfate,) and a derivatizing reagent (OPA, o-phthalaldehyde; NAC, N-acetylcysteine), histamine in the mast cells was detected at high-sensitivity level without further procedures. During ICD-HPCE, the mast cells injected in the capillary were lysed with the lysing reagent, free histamine released from the cell was labeled with the derivatizing reagent, and its derivative was electromigrated, separated and detected with a fluorescence detector (excitation wavelength at 340 nm, emission wavelength at 450 nm) in a fused-silica capillary (75 cm x effective length x 50 microm I.D.). The run buffer used was a 20 mM phosphate-borate buffer (pH 10) containing 20 mM SDS, 2 mM OPA and 2 mM NAC. This method was also examined with regard to the possibility of its use for determination of histamine at the single mast cell level. PMID- 10677008 TI - Characterization of a solid-phase extraction device for discontinuous on-line preconcentration in capillary electrophoresis-based peptide mapping. AB - Peptide mapping by capillary electrophoresis (CE) with UV detection is problematic for the characterization of proteins that can only be obtained at low micromolar concentrations. Dilution of peptide fragments during digestion of the protein can further reduce the detection sensitivity in peptide mapping to the point where analysis at sub-micromolar concentrations is not possible. A remedy to this problem is preconcentration (sample enrichment) of the proteolytic digest by solid-phase extraction (SPE). To minimize non-specific adsorptive losses during sample handling, on-line SPE-CE is preferred. However, packed-inlet SPE-CE is not always feasible due to either instrument or sample limitations. We describe here a simple method of preconcentration by discontinuous on-line SPE CE, specifically applied to peptide mapping in low-pH separation buffer after protein digestion in a solid-phase enzyme microreactor. The SPE-CE system does not require application of a low pressure during electrophoretic separation to overcome reversed electroosmotic flow because the preconcentrator device is disconnected from the separation capillary before the electric field is applied. Up to a 500-fold preconcentration factor can be achieved with this device, which can be reused for many samples. Parameters such as the volume of desorption solution, the adsorption/desorption (chromatographic) process, reproducibility of packing the SPE preconcentrator and effects of sample concentration on the peptide map are investigated. PMID- 10677009 TI - High-performance thin-layer chromatographic determination of theophylline in plasma. AB - A high-performance thin-layer chromatographic method for quantification of theophylline from plasma is described. The calibration curves of theophylline in methanol and in plasma were linear in the range 20-100 ng. The correlation coefficients were 0.9971+/-0.0011 and 0.9955+/-0.0003 for standard curves in methanol and in plasma, respectively. The limit of quantitation of theophylline in human plasma (assay sensitivity) was 20 ng and no interference from endogenous compounds was observed. The recovery of theophylline from human plasma using the described assay procedure was 89%. The mean relative standard deviations for intra- and inter-day analyses were 1.67% and 2.34% for 50 ng and 2.25% and 3.14% for 75 ng theophylline concentration, respectively. The method was utilized to monitor plasma concentration of theophylline post-administration of sustained release tablets in human patient volunteers. PMID- 10677010 TI - Determination of lamotrigine in human serum by liquid chromatography. AB - A rapid high-performance liquid chromatographic method was developed using a short silica column (30 mm x 4.6 mm) with an aqueous methanol mobile phase consisting of methanol-water-NH4H2PO4 (94:5.96:0.04) adjusted to a final apparent pH of 5.0 and pumped at a flow-rate of 1 ml/min. Ultraviolet detection was carried out at a wavelength of 280 nm, and serum samples were prepared for HPLC analysis by extraction into dichloromethane after basification. Lamotrigine was eluted at 0.96 min. Within-day variation of the method was 4.46% at 0.75 microg/ml and 2.37% at 6.0 microg/ml, and day-to-day variation was 9.10% at 0.75 microg/ml and 7.28% at 6.0 microg/ml. PMID- 10677011 TI - Simple and efficient method for the detection of diethylenetriaminepentaacetic acid. AB - Diethylenetriaminepentaacetic acid (DTPA) is a commonly used chelating agent. Its antiviral, antibacterial and immunomodulatory effects are well documented. DTPA forms a highly stable complex with lead (II) with an increased absorption coefficient and a bathochromic shift of the absorption maximum compared to pure DTPA. Based on this complex a high-performance liquid chromatographic method for the quantitative detection of DTPA in biological fluids was developed. A calibration curve was prepared and linearity was shown in the concentration range between 10 mg l(-1) and 1000 mg l(-1) DTPA. The recovery in water and in human plasma showed the method to be suitable for routine use. PMID- 10677012 TI - Temporal and spatial expression of the period gene in the reproductive system of the codling moth. AB - The authors examined patterns of spatial and temporal expression of Drosophila per gene homologue in the codling moth, Cydia pomonella. Since sperm release in moths is regulated in a circadian manner by an autonomous clock that is independent from the brain, the authors investigated per expression in male reproductive system along with its expression in moth heads. per mRNA is rhythmically expressed with the same phase and amplitude in both tissues under light-dark (LD) conditions. The levels of per mRNA are low during the day, start to increase before lights-off, reach the peak in dark, and decrease after lights on. In constant darkness (DD), cycling of per mRNA continued in heads with severely blunted amplitude. No cycling of per mRNA was detected in testis in DD. In situ hybridization and immunocytochemistry revealed distinct spatial patterns of per expression in the moth reproductive system. There is no expression of per in cells forming the wall of testes or in sperm bundles. However, per mRNA and protein are rhythmically expressed in the epithelial cells forming the wall of the upper vas deferens (UVD) and in the cells of the terminal epithelium, which are involved in the circadian gating of sperm release. Increase in per mRNA in the UVD coincides with sperm accumulation in this part of the insect reproductive system. PMID- 10677013 TI - Involvement of the period gene in developmental time-memory: effect of the perShort mutation on phase shifts induced by light pulses delivered to Drosophila larvae. AB - Phases of circadian locomotor activity rhythms of adult Drosophila reared in constant darkness have been shown to be set by a light stimulus delivered as early as the first-instar larval stage. This implies that a circadian clock functions continuously throughout postembryonic development. The clock genes period (per) and timeless (tim) are expressed cyclically in the larval central nervous system of Drosophila, and daily oscillations of per expression persist throughout metamorphosis in a group of cells, which gives rise to the pacemaker cells underlying locomotor activity rhythms of adults. Therefore, PER and TIM cyclings in these neurons may be responsible for the phenomenon of "larval time memory." In the absence of any evidence for the involvement of these genes in such a developmental clock, and because circadian-pacemaker functions are underanalyzed in terms of the functions during development, the authors tested the time-memory of a fast-clock period mutant. They show that dark-reared perS mutant individuals as well as wild-type flies can be entrained as larvae and that a brief light pulse given to such entrained larvae can induce phase shifts in animals of either genotype. However, the direction and magnitude of phase shifts were different between wild type and perS, suggesting that a clock under the control of period gene participates in the regulation of developmental time memory. The authors show that the relevant clock can be entrained by two light input pathways, one involving the phospholipase C encoded by the norpA gene, the other mediated by the blue-light receptor cryptochrome. Phase shifts of molecular oscillations during the larval stage were smaller than those measured by adult behavior, suggesting molecularly transient responses during development. PMID- 10677014 TI - Physiological response properties of cat retinal ganglion cells projecting to suprachiasmatic nucleus. AB - The aim of this experiment was to characterize the physiological properties of cat retinal ganglion cells that project to the suprachiasmatic nucleus (SCN). Retrogradely labeled SCN-projecting ganglion cells were recorded extracellularly in vitro. For the first time, this study provides crucial information on visual response properties of ganglion cells in the entrainment circuitry. All recorded cells gave sustained responses (n = 9). Although most of the cells (n = 8) had an "on" center receptive field, one cell showed "on-off" center receptive field properties. The range of receptive field sizes was 2 to 5 deg. For most of the cells tested, the spectral wavelength that evoked peak responses was 500 nm (3 out of 5 cells). All recorded cells (n = 9) preferred still or extremely slow moving stimuli (3.3 deg/s). These results indicate that cat SCN-projecting cells receive inputs from conventional photoreceptors. The hypothesis that both conventional and cryptochromic photoreceptors are involved in transferring photic signals is discussed. PMID- 10677015 TI - The suprachiasmatic nucleus in organotypic slice cultures of the common vole (Microtus arvalis): comparison of development with rat and hamster and the effect of age. AB - The intrinsic properties of the suprachiasmatic nucleus (SCN), the site of the main circadian pacemaker in mammals, have recently been studied in vitro by means of organotypic slice culturing. So far, only neonatal rats and mice have been used for such developmental and functional analyses of the isolated pacemaker. Here, the authors present a comparative developmental study of the SCN of voles, rats, and hamsters in organotypic slice cultures. In contrast to strictly circadian organization of behavior in rats and hamsters, common voles (Microtus arvalis) are characterized by large variability in the strength of circadian organization of behavior. It is not known to what extent this variability is reflected in the intrinsic features of the SCN. Cultures were prepared from rat, hamster, and vole pups (6 to 9 days old) for the purpose of species comparison. In addition, the authors studied the relation between age and development in cultures from pup (7 to 10 days old), juvenile (15 to 16 days old), and young adult (1 to 2 months old) voles. In contrast to the situation in rat and hamster, the most striking feature in neonatal voles is the variability in shape of the final, fully developed culture and its poor resemblance with the in vivo SCN. The SCN of adult voles, however, could be cultured successfully while retaining its morphological organization seen in situ. Phase-contrast microscopy and immunocytochemical staining for vasopressin and glial fibrillary acidic protein revealed that cultures of pup and juvenile voles still have potential for neurogenesis and morphological reorganization. Young voles, therefore, can serve as a model to study the developmental establishment of a functional circadian pacemaker, while adult voles allow the study of intrinsic pacemaker properties in relation to previously recorded behavior of the donor and aging-related pacemaker dysfunction. PMID- 10677016 TI - Exogenous melatonin reduces the resynchronization time after phase shifts of a nonphotic zeitgeber in the house sparrow (Passer domesticus). AB - Continuous melatonin administration via silastic implants accelerates the resynchronization of the circadian locomotor activity rhythm in house sparrows (Passer domesticus) after exposure to phase shifts of a weak light-dark cycle. Constant melatonin might induce this effect either by increasing the sensitivity of the visual system to a light zeitgeber or by reducing the degree of self sustainment of the circadian pacemaker. To distinguish between these two possible mechanisms, two groups of house sparrows, one carrying melatonin implants and the other empty implants, were kept in constant dim light and subjected to advance and delay shifts of a 12-h feeding phase. The resynchronization times of their circadian feeding rhythm following the phase shifts were significantly shorter when the birds carried melatonin implants than when they carried empty implants. In a second experiment, melatonin-implanted and control birds were released into food ad libitum conditions 2 days after either a delay or an advance phase shift. The number of hours by which the activity rhythms had been shifted on the second day in food ad libitum conditions was assessed. Melatonin-implanted house sparrows had significantly larger phase shifts in their circadian feeding rhythm than control birds. This is in accordance with the first experiment since a larger phase shift at a given time reflects accelerated resynchronization. Additionally, the second experiment also excludes any possible masking effects of the nonphotic zeitgeber. In conclusion, constant melatonin accelerates resynchronization even after phase shifts of a nonphotic zeitgeber, indicating that constant high levels of melatonin can reduce the degree of self-sustainment of the circadian pacemaker independent of any effects on the photoreceptive system. PMID- 10677017 TI - Restricted daytime feeding attenuates reentrainment of the circadian melatonin rhythm after an 8-h phase advance of the light-dark cycle. AB - It is well established that in the absence of photic cues, the circadian rhythms of rodents can be readily phase-shifted and entrained by various nonphotic stimuli that induce increased levels of locomotor activity (i.e., benzodiazepines, a new running wheel, and limited food access). In the presence of an entraining light-dark (LD) cycle, however, the entraining effects of nonphotic stimuli on (parts of) the circadian oscillator are far less clear. Yet, an interesting finding is that appropriately timed exercise after a phase shift can accelerate the entrainment of circadian rhythms to the new LD cycle in both rodents and humans. The present study investigated whether restricted daytime feeding (RF) (1) induces a phase shift of the melatonin rhythm under entrained LD conditions and (2) accelerates resynchronization of circadian rhythms after an 8 h phase advance. Animals were adapted to RF with 2-h food access at the projected time of the new dark onset. Before and at several time points after the 8-h phase advance, nocturnal melatonin profiles were measured in RF animals and animals on ad libitum feeding (AL). In LD-entrained conditions, RF did not cause any significant changes in the nocturnal melatonin profile as compared to AL. Unexpectedly, after the 8-h phase advance, RF animals resynchronized more slowly to the new LD cycle than AL animals. These results indicate that prior entrainment to a nonphotic stimulus such as RF may "phase lock" the circadian oscillator and in that way hinder resynchronization after a phase shift. PMID- 10677018 TI - Probing the circadian pacemaker of a mouse using two light pulses. AB - In two separate sets of experiments, the phases of the locomotor activity rhythm of the nocturnal field mouse Mus booduga were probed using two light pulses (LPs). In the first set of experiments, the circadian pacemaker underlying the locomotor activity rhythm was perturbed at circadian time 14 (CT 14) using a resetting light pulse LP1 of 1000 lux intensity and 15 min duration. The phases of the resetting pacemaker were then probed at all even CTs between CT 16 and CT 14 using a PRC probing light pulse LP2 of equal strength. The "LP2 PRC" thus obtained was then compared with the single light pulse PRC in terms of the area under delay (D) and advance (A) zones of the PRCs. The time course and waveform of the two LP PRCs suggest that the LP2 PRC resembled the single LP PRC, displaced by 2 h toward the right. The LP1 PRC had smaller D compared to the single LP PRC (p = 0.007), whereas both the PRCs had A of equal magnitude (p = 0.23). This suggests that the pacemaker phase shifts rapidly after LP perturbations. In the second set of experiments, the LP1 was administered at CT 14. The phase of the pacemaker was then perturbed on day 1 (next cycle after LP1) either 2 h after activity onset (at ca. CT 14 of the transient cycle) or 8 h after activity onset (at ca. CT 20 of the transient cycle) using an LP2 of equal strength. It was observed that the steady-state phase shifts evoked by positioning an LP2, 2 h after activity onset, were positively correlated with the phase shifts observed on day 1. The steady-state phase shifts observed, when the LP2 was positioned, 8 h after activity onset, were negatively correlated with the phase shifts observed on day 1. These results suggest that the transient cycles do not mirror the state of the pacemaker oscillator. PMID- 10677019 TI - Identifying components of Max Factor. PMID- 10677020 TI - Getting the membrane into shape for endocytosis. PMID- 10677021 TI - Siesta-time is in the genes. PMID- 10677022 TI - The binding problem. PMID- 10677023 TI - The psychophysical evidence for a binding problem in human vision. PMID- 10677024 TI - The role of neural mechanisms of attention in solving the binding problem. PMID- 10677025 TI - The temporal correlation hypothesis of visual feature integration: still alive and well. PMID- 10677026 TI - Neuronal synchrony: a versatile code for the definition of relations? PMID- 10677027 TI - Synchrony unbound: a critical evaluation of the temporal binding hypothesis. PMID- 10677028 TI - Specialized representations in visual cortex: a role for binding? PMID- 10677029 TI - Are cortical models really bound by the "binding problem"? PMID- 10677030 TI - The what and why of binding: the modeler's perspective. PMID- 10677031 TI - Solutions to the binding problem: progress through controversy and convergence. PMID- 10677032 TI - BMPs as mediators of roof plate repulsion of commissural neurons. AB - During spinal cord development, commissural (C) neurons, located near the dorsal midline, send axons ventrally and across the floor plate (FP). The trajectory of these axons toward the FP is guided in part by netrins. The mechanisms that guide the early phase of C axon extension, however, have not been resolved. We show that the roof plate (RP) expresses a diffusible activity that repels C axons and orients their growth within the dorsal spinal cord. Bone morphogenetic proteins (BMPs) appear to act as RP-derived chemorepellents that guide the early trajectory of the axons of C neurons in the developing spinal cord: BMP7 mimics the RP repellent activity for C axons in vitro, can act directly to collapse C growth cones, and appears to serve an essential function in RP repulsion of C axons. PMID- 10677033 TI - Endophilin/SH3p4 is required for the transition from early to late stages in clathrin-mediated synaptic vesicle endocytosis. AB - Endophilin/SH3p4 is a protein highly enriched in nerve terminals that binds the GTPase dynamin and the polyphosphoinositide phosphatase synaptojanin, two proteins implicated in synaptic vesicle endocytosis. We show here that antibody mediated disruption of endophilin function in a tonically stimulated synapse leads to a block in the invagination of clathrin-coated pits adjacent to the active zone and therefore to a block of synaptic vesicle recycling. We also show that in a cell-free system, endophilin is not associated with clathrin coats and is a functional partner of dynamin. Our findings suggest that endophilin is part of a biochemical machinery that acts in trans to the clathrin coat from early stages to vesicle fission. PMID- 10677034 TI - Integration of NPY, AGRP, and melanocortin signals in the hypothalamic paraventricular nucleus: evidence of a cellular basis for the adipostat. AB - Energy stores are held relatively constant in many mammals. The circuitry necessary for maintaining energy homeostasis should (1) sense the amount of energy stored in adipose tissue, (2) sense and integrate the multiple opposing signals regarding nutritional state, and (3) provide output regulating energy intake and expenditure to maintain energy homeostasis. We demonstrate that individual neurons within the paraventricular nucleus of the hypothalamus (PVH) are capable of detection and integration of orexigenic (neuropeptide Y [NPY]) and anorexigenic (melanocortin) signals, that NPY and melanocortins are functional antagonists of each other within the PVH in the regulation of feeding behavior, and that melanocortin administration within the PVH regulates both feeding behavior and energy expenditure. These data provide a cellular basis for the adipostat within neurons in the PVH that appear to be jointly regulated by NPY- and melanocortin-responsive neurons. PMID- 10677035 TI - Identification of maxillary factor, a maxillary process-derived chemoattractant for developing trigeminal sensory axons. AB - Trigeminal sensory axons project to several epithelial targets, including those of the maxillary and mandibular processes. Previous studies identified a chemoattractant activity, termed Maxillary Factor, secreted by these processes, which can attract developing trigeminal axons in vitro. We report that Maxillary Factor activity is composed of two neurotrophins, neurotrophin-3 (NT-3) and Brain Derived Neurotrophic Factor (BDNF), which are produced by both target epithelium and pathway mesenchyme and which are therefore more likely to have a trophic effect on the neurons or their axons than to provide directional information, at least at initial stages of trigeminal axon growth. Consistent with this, the initial trajectories of trigeminal sensory axons are largely or completely normal in mice deficient in both BDNF and NT-3, indicating that other cues must be sufficient for the initial stages of trigeminal axon guidance. PMID- 10677036 TI - Learned movements elicited by direct stimulation of cerebellar mossy fiber afferents. AB - Definitive evidence is presented that the conditioned stimulus (CS) in classical conditioning reaches the cerebellum via the mossy fiber system. Decerebrate ferrets received paired forelimb and periocular stimulation until they responded with blinks to the forelimb stimulus. When direct mossy fiber stimulation was then given, the animals responded with conditioned blinks immediately, that is, without ever having been trained to the mossy fiber stimulation. Antidromic activation was prevented by blocking mossy fibers with lignocaine ventral to the stimulation site. It could be excluded that cerebellar output functioned as the CS. Analysis of latencies suggests that conditioned responses (CRs) are not generated by mossy fiber collaterals to the deep nuclei. Hence, the memory trace is probably located in the cerebellar cortex. PMID- 10677037 TI - Differential processing of objects under various viewing conditions in the human lateral occipital complex. AB - The invariant properties of human cortical neurons cannot be studied directly by fMRI due to its limited spatial resolution. Here, we circumvented this limitation by using fMR adaptation, namely, reduction of the fMR signal due to repeated presentation of identical images. Object-selective regions (lateral occipital complex [LOC]) showed a monotonic signal decrease as repetition frequency increased. The invariant properties of fMR adaptation were studied by presenting the same object in different viewing conditions. LOC exhibited stronger fMR adaptation to changes in size and position (more invariance) compared to illumination and viewpoint. The effect revealed two putative subdivisions within LOC: caudal-dorsal (LO), which exhibited substantial recovery from adaptation under all transformations, and posterior fusiform (PF/LOa), which displayed stronger adaptation. This study demonstrates the utility of fMR adaptation for revealing functional characteristics of neurons in fMRI studies. PMID- 10677038 TI - Effects of lexicality, frequency, and spelling-to-sound consistency on the functional anatomy of reading. AB - Functional neuroimaging was used to investigate three factors that affect reading performance: first, whether a stimulus is a word or pronounceable non-word (lexicality), second, how often a word is encountered (frequency), and third, whether the pronunciation has a predictable spelling-to-sound correspondence (consistency). Comparisons between word naming (reading) and visual fixation scans revealed stimulus-related activation differences in seven regions. A left frontal region showed effects of consistency and lexicality, indicating a role in orthographic to phonological transformation. Motor cortex showed an effect of consistency bilaterally, suggesting that motoric processes beyond high-level representations of word phonology influence reading performance. Implications for the integration of these results into theoretical models of word reading are discussed. PMID- 10677039 TI - How a circadian clock adapts to seasonal decreases in temperature and day length. AB - We show that a thermosensitive splicing event in the 3' untranslated region of the mRNA from the period (per) gene plays an important role in how a circadian clock in Drosophila adapts to seasonally cold days (low temperatures and short day lengths). The enhanced splicing of this intron at low temperatures advances the steady state phases of the per mRNA and protein cycles, events that significantly contribute to the preferential daytime activity of flies on cold days. Because the accumulation of PER is also dependent on the photosensitive TIMELESS (TIM) protein, long photoperiods partially counteract the cold-induced advances in the oscillatory mechanism by delaying the daily increases in the levels of TIM. Our findings also indicate that there is a temperature-dependent switch in the molecular logic governing cycles in per mRNA levels. PMID- 10677040 TI - Serotonin inhibition of synaptic transmission: Galpha(0) decreases the abundance of UNC-13 at release sites. AB - We show that serotonin inhibits synaptic transmission at C. elegans neuromuscular junctions, and we describe a signaling pathway that mediates this effect. Release of acetylcholine from motor neurons was assayed by measuring the sensitivity of intact animals to the acetylcholinesterase inhibitor aldicarb. By this assay, exogenous serotonin inhibited acetylcholine release, whereas serotonin antagonists stimulated release. The effects of serotonin on synaptic transmission were mediated by GOA-1 (a Galpha0 subunit) and DGK-1 (a diacylglycerol [DAG] kinase), both of which act in the ventral cord motor neurons. Mutants lacking goa 1 G(alpha)0 accumulated abnormally high levels of the DAG-binding protein UNC-13 at motor neuron nerve terminals, suggesting that serotonin inhibits synaptic transmission by decreasing the abundance of UNC-13 at release sites. PMID- 10677041 TI - Retention of supraspinal delta-like analgesia and loss of morphine tolerance in delta opioid receptor knockout mice. AB - Gene targeting was used to delete exon 2 of mouse DOR-1, which encodes the delta opioid receptor. Essentially all 3H-[D-Pen2,D-Pen5]enkephalin (3H-DPDPE) and 3H [D-Ala2,D-Glu4]deltorphin (3H-deltorphin-2) binding is absent from mutant mice, demonstrating that DOR-1 encodes both delta1 and delta2 receptor subtypes. Homozygous mutant mice display markedly reduced spinal delta analgesia, but peptide delta agonists retain supraspinal analgesic potency that is only partially antagonized by naltrindole. Retained DPDPE analgesia is also demonstrated upon formalin testing, while the nonpeptide delta agonist BW373U69 exhibits enhanced activity in DOR-1 mutant mice. Together, these findings suggest the existence of a second delta-like analgesic system. Finally, DOR-1 mutant mice do not develop analgesic tolerance to morphine, genetically demonstrating a central role for DOR-1 in this process. PMID- 10677042 TI - A novel nociceptor signaling pathway revealed in protein kinase C epsilon mutant mice. AB - There is great interest in discovering new targets for pain therapy since current methods of analgesia are often only partially successful. Although protein kinase C (PKC) enhances nociceptor function, it is not known which PKC isozymes contribute. Here, we show that epinephrine-induced mechanical and thermal hyperalgesia and acetic acid-associated hyperalgesia are markedly attenuated in PKCepsilon mutant mice, but baseline nociceptive thresholds are normal. Moreover, epinephrine-, carrageenan-, and nerve growth factor- (NGF-) induced hyperalgesia in normal rats, and epinephrine-induced enhancement of tetrodotoxin-resistant Na+ current (TTX-R I(Na)) in cultured rat dorsal root ganglion (DRG) neurons, are inhibited by a PKCepsilon-selective inhibitor peptide. Our findings indicate that PKCepsilon regulates nociceptor function and suggest that PKCepsilon inhibitors could prove useful in the treatment of pain. PMID- 10677043 TI - Activation of a TRPC3-dependent cation current through the neurotrophin BDNF. AB - Nonvoltage-gated cation currents, which are activated following stimulation of phospholipase C (PLC), appear to be major modes for Ca2+ and Na+ entry in mammalian cells. The TRPC channels may mediate some of these conductances since their expression in vitro leads to PLC-dependent cation influx. We found that the TRPC3 protein was highly enriched in neurons of the central nervous system (CNS). The temporal and spatial distribution of TRPC3 paralleled that of the neurotrophin receptor TrkB. Activation of TrkB by brain-derived nerve growth factor (BDNF) led to production of a PLC-dependent, nonselective cation conductance in pontine neurons. Evidence is provided that TRPC3 contributes to this current in vivo. Thus, activation of TrkB and PLC leads to a TRPC3-dependent cation influx in CNS neurons. PMID- 10677044 TI - Nuclear accumulation of truncated atrophin-1 fragments in a transgenic mouse model of DRPLA. AB - Dentatorubral and pallidoluysian atrophy (DRPLA) is a member of a family of progressive neurodegenerative diseases caused by polyglutamine repeat expansion. Transgenic mice expressing full-length human atrophin-1 with 65 consecutive glutamines exhibit ataxia, tremors, abnormal movements, seizures, and premature death. These mice accumulate atrophin-1 immunoreactivity and inclusion bodies in the nuclei of multiple populations of neurons. Subcellular fractionation revealed 120 kDa nuclear fragments of mutant atrophin-1, whose abundance increased with age and phenotypic severity. Brains of DRPLA patients contained apparently identical 120 kDa nuclear fragments. By contrast, mice overexpressing atrophin-1 with 26 glutamines were phenotypically normal and did not accumulate the 120 kDa fragments. We conclude that the evolution of neuropathology in DRPLA involves proteolytic processing of mutant atrophin-1 and nuclear accumulation of truncated fragments. PMID- 10677045 TI - Infantile colic: aetiology and prognosis. PMID- 10677046 TI - Towards a "new beginning": dietary fat restrictions in infancy? PMID- 10677047 TI - Long-term coping in childhood cancer survivors--influence of illness, treatment and demographic factors. PMID- 10677048 TI - Using a large-scale screening method to detect language disability in three-year olds. PMID- 10677049 TI - Does breastfeeding increase thymus size? PMID- 10677050 TI - Infantile colic. Follow-up at four years of age: still more "emotional". AB - This paper presents a follow-up at 4 y of formerly colicky infants and controls, with respect to behaviour, temperament, eating and sleeping habits, psychosomatic complaints, number of hospital stays, growth and "family climate". There were no differences between the two groups in most parameters studied. However, ex colicky children displayed more negative emotions according to the temperament scale. There were also more negative moods during meals, and more reported stomach-ache. Although relationships regarding crying and mother-infant interaction remain extremely complex, the findings point toward a possible temperamental contribution to the pathogenesis of the infantile colic syndrome. PMID- 10677051 TI - Effectiveness of casein hydrolysate feedings in infants with colic. AB - This study found that two casein hydrolysate formulas varying in composition were equally effective in managing colicky symptoms associated with protein sensitivity. Both hydrolysate formulas were associated with a significant, comparable reduction in crying duration and intensity from baseline in 15 of 22 infants with complete data. Subsequent challenge data suggest that the population studied were infants experiencing colicky symptoms due to protein sensitivity. A greater proportion of infants showed a positive reaction (> or = 1.5 h of crying/d) to the protein challenges than the placebo challenge, and crying was rated as more intense during whey and milk protein challenges. PMID- 10677052 TI - Treatment of infant colic with amino acid-based infant formula: a preliminary study. AB - Infant colic, a common disorder of infancy, is characterized by excessive crying and fussing. In this preliminary study we examined whether Neocate, an amino acid based formula, would be accepted by formula-fed infants with colic, 3-7 wk of age, and whether Neocate would improve their symptoms. Six infants with colic were studied using Barr-type infant behavior diaries for 3-6 d on their current formula and then for 5-17 d on Neocate exclusively. All infants tolerated Neocate well and all improved, usually within 1-2 d. The total time spent crying and fussing was reduced by an average of 45%, representing a decrease of 1.0 to 5.2 h daily. After colic symptoms improved, infants were challenged with oral doses of 75 mg of bovine IgG at a 1 mg/ml concentration in order to assess its potential role in colic. Bovine IgG challenges resulted in increased crying and fussing behavior, suggesting that this protein may be etiologically important. PMID- 10677053 TI - Fat intake and metabolism in Swedish and Italian infants. AB - The purpose of the study was to compare fat intake and metabolism between two infant populations from Sweden and Italy given breast milk or similar infant formulas, but different weaning foods. Nutrient intake and fat metabolism were studied prospectively from 3-12 mo in 68 Swedish and 46 Italian healthy infants, breastfed or given similar infant formulas in combination with Swedish or Mediterranean weaning foods. Although nutrient intake and fat metabolism were similar at 6 mo, fat intake was lower at 12 mo in the Italian than in the Swedish formula group (p < 0.001). At 6 and 12 mo, higher dietary ratios of monounsaturated to saturated fatty acids (p < 0.01 and p < 0.001, respectively), and monounsaturated to polyunsaturated fatty acids (p < 0.05, p < 0.001) were found in the Italian than in the Swedish formula group. Total cholesterol and apolipoprotein B were lower at 6 mo (p < 0.01) in Italian breastfed infants than in Swedish ones. Lower concentrations at 6 and 12 mo of total cholesterol (p < 0.05, p < 0.05, respectively), apolipoprotein B (p < 0.05, p < 0.01) and triglycerides (p < 0.001, p < 0.01), and of apolipoprotein A1 (p < 0.01) at 12 mo, were found in the Italian formula group than in the Swedish one. In conclusion, plasma total cholesterol, apolipoprotein B and triglycerides were found to be lower in Italian infants than in Swedish infants during the second half of infancy. These findings may partly result from differences in fat compositions between Swedish and Mediterranean weaning diets and in total fat intake in late infancy. Differences in duration of breastfeeding and possibly in breast milk composition may also have influenced our results. PMID- 10677054 TI - Prediction of growth response in prepubertal children treated with growth hormone for idiopathic growth hormone deficiency. AB - Several multiple regression models have been developed to predict the first-year growth response to human growth hormone (hGH) in children with growth hormone deficiency (GHD). It was the aim of this study to analyse the significance of various growth parameters for a height prediction model. Data from 148 prepubertal children with idiopathic GHD were evaluated. The prediction model was developed by means of univariate and stepwise linear regression analysis and an "all possible" regression approach using Mallow's C(p) statistics. Six out of eight selected variables had a significant influence on the first-year growth rate. The most important parameter was the difference between target height SDS and height SDS at the start of therapy (THSDS-HSDSCO), accounting for 23.95% and 25.74% of the variability. No other single variable or combination of variables was more informative than the variable THSDS-HSDSCO alone. From these data, growth velocity for the first year of hGH treatment was estimated as 1.106 (THSDS HSDSCO) + 6.8 cm/y +/- 2.2 cm (SE), allowing a prediction for different intervals between THSDS and HSDSCO. This equation was validated in a small group of 18 GHD patients demonstrating a predicted vs. observed first-year growth rate of 9.4 +/- 1.1 vs. 9.5 +/- 2.6 cm/y. We conclude that the difference between THSDS and height SDS at the start of therapy is an important predictor of the first-year growth response in children treated with hGH for idiopathic GHD. Unlike in previous studies, additional parameters did not increase predictability. PMID- 10677055 TI - Comparison of the growth-promoting effects of insulin-like growth factor I and growth hormone in the early years of life. AB - AIMS: Four infants with isolated growth hormone deficiency (IGHD) and five with Laron syndrome (LS) were studied. Birth length ranged from -2.5 to -4.5 SDS in both groups. RESULTS: Untreated IGHD children decreased in length from -2.5 to 5.2 SDS at 1 y and to -5.7 SDS at 2 y. Human growth hormone (hGH) treatment (0.07 U/kg/d) increased height by 1.2-2.4 SDS in 3 y. Untreated children with LS decreased in length from -3.5 to -6.5 SDS. Insulin-like growth factor (IGF)-I treatment (150-200 microg/kg/d) in 3 LS patients increased height by 0.5-1.5 SDS in 3 y. All untreated infants had borderline or below normal head circumferences. Both treatments induced a rapid catch-up in head size. In the two untreated LS patients, head circumference remained subnormal. CONCLUSIONS: Despite similar birth length, infants with IGHD responded better to hGH in terms of linear growth than did infants with LS to IGF-I, whereas the response in brain growth was similar. PMID- 10677056 TI - Association of serum low-density lipoprotein metabolism with oestrogen receptor gene polymorphisms in healthy children. AB - The aim of this study was to reveal the association of serum lipid and apolipoprotein levels with oestrogen receptor (ER) Xba I and Pvu II polymorphisms in 102 healthy Japanese school children (56M, 46F) aged 10-15 y. Each genotype of the genomic DNA extracted from peripheral leukocytes was determined using polymerase chain reaction and digestion with Xba I or Pvu II. The genotypes were coded as either X1 or X2 (Xba I) and P1 or P2 (Pvu II), when XI, P1 signified the absence of and X2, P2 the presence of restriction sites. The fasting serum total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein (LDL) cholesterol, triglyceride and apolipoproteins A1, B and E were measured. In the Xba I polymorphism, LDL cholesterol, apolipoprotein B levels of the XI/XI genotype were significantly higher than those of the others. The other lipid and apolipoprotein levels were not significantly different among the three genotypes. In the Pvu II polymorphism, there were no significant differences in serum lipids and lipoproteins among the three genotypes. This study reveals that Xba I polymorphisms are related to LDL metabolism. These findings support previous reports that the LDL-lowering effects of oestrogen occur through the ER (alpha) pathway. The Xba I polymorphism may be one of the genetic factors in the control of LDL metabolism. PMID- 10677057 TI - Islet cell antibody frequency differs from that of glutamic acid decarboxylase antibodies/IA2 antibodies after diagnosis of diabetes. AB - The combination of glutamic acid decarboxylase (GAD) 65 antibodies (GADA) and protein tyrosine phosphatase-like protein IA2 antibodies (IA2-ab), measured by radioligand binding assays, has been suggested to replace islet cell antibodies (ICA), measured by indirect immunofluorescence, as a marker for autoimmune type I diabetes. The aim of this study was to compare the frequency of ICA and GADA and/or IA2-ab not only at, but also after the diagnosis of diabetes. ICA, GADA and IA2-ab were therefore assessed at and up to 11 y after the diagnosis of diabetes in 86 children (1-15-y-old). At diagnosis, ICA were found in 74 (86%) and GADA and/or IA2-ab in 79 (92%) of the diabetic children. Hence, there was no major difference in frequency between ICA and GADA and/or IA2-ab at diagnosis of diabetes. At follow-up, however, ICA were less frequent than GADA and/or IA2-ab; 1-3 y after diagnosis ICA were found in 12 (44%) and GADA and/or IA2-ab in 24 (89%) of 27 children (p=0.001); 4-6 y after diagnosis ICA were found in 7 (24%) and GADA and/or IA2-ab in 27 (93%) of 29 children (p < 0.0001); 7-11 y after diagnosis ICA were found in 4 (13%) and GADA and/or IA2-ab in 21 (70%) of 30 children (p < 0.0001). We conclude that the frequency of ICA does not always correspond to that of GADA and/or IA2-ab. Many years after diagnosis of diabetes, measurements of GADA and IA2-ab, but not ICA, detect autoimmunity in high frequency. PMID- 10677058 TI - Visual event-related potentials in children with phenylketonuria. AB - Visual event-related potentials (ERPs) were examined in 16 children (aged 5-14 y) with phenylketonuria (PKU) and 16 age- and sex-matched controls. Lifetime median measures of phenylalanine (Phe) were 230-460 micromol/l. The most recent Phe levels were 56-624 micromol/l. ERPs were recorded whilst the children performed a discrimination task. All stimuli were square wave gratings degree, which appeared for 33 ms. A response to an infrequent grating that differed in orientation or spatial frequency was required. The older children with PKU had a delay in the first peak (P1) of the ERP, and age-related changes in the amplitude of P1. There was attenuation of the second peak across age groups in PKU. There was no evidence of reduced response accuracy or longer reaction times in children with PKU. Latencies of the cognitive P3 were not delayed in PKU. The delayed early peaks are consistent with previous studies that have shown delayed visual evoked potentials in PKU. The lack of differences in reaction time and P3 may be due to relatively good Phe control in children with PKU, or to the simplicity of the task. Suggestions are made for future ERP studies of PKU. PMID- 10677059 TI - Gait disturbance interpreted as cerebellar ataxia after MMR vaccination at 15 months of age: a follow-up study. AB - Measles, mumps and rubella (MMR) vaccination was included in the Danish childhood vaccination programme in 1987. During the following 10-y period, 550 notification records of adverse events after MMR vaccination at 15 mo of age have been registered, and a total of 41 notifications have included "gait disturbance". This corresponds to a frequency of 8 per 100,000 doses of MMR vaccine used for 15 mo-old children. The symptoms and signs are characteristic of cerebellar ataxia. In 28 notifications, the descriptions by the doctors included only "gait disturbance", while in 13 an additional interpretation was included. Thirty-two parents (78%) filled in a questionnaire and 26 (63%) agreed to participate in a clinical follow-up study. The gait disturbance symptoms mainly occurred 7-14 d after the vaccination, and the duration was median 1-2 wk (range 1 d to more than 4 mo). One-third of the children had symptoms lasting more than 2 wk. Significantly more children with long duration of symptoms had some kind of complaint or clinical signs at the follow-up in 1997. Gait disturbance registered after MMR vaccination seems to be more frequent than hitherto reported. Most cases are mild and short-lasting and a longer duration of symptoms seems to be predictive of late sequelae. A clinical diagnosis of cerebellar ataxia after MMR and the exact frequency of this adverse event remains to be tested in prospective studies. PMID- 10677060 TI - Magnetocardiographic determination of the developmental changes in PQ, QRS and QT intervals in the foetus. AB - In order to determine developmental changes in atrioventricular (PQ), ventricular depolarizing (QRS) and QT intervals of the foetal heart, we recorded foetal magnetocardiographic waveforms using a superconducting quantum interference device system in a magnetically shielded room in 150 uncomplicated foetuses of gestational age >20 wk. Recording of the QRS waveform was successful in 128 (85%) of the subjects, based on unaveraged tracings. After signal averaging of the data from these 128 cases, P waves were recognized in 102 (68%) subjects and T waves in 64 (43%). The QRS interval, ranging from 32-74 ms, showed a positive linear correlation with the gestational age, which probably reflects an increase in the number and size of myocardial cells. The PQ interval showed low correlation with the gestational age, and was rather constant, with an average value of 100 ms. The QT interval ranged from 180-302 ms, and tended to be slightly shorter during early gestation. Although the success rate of measuring the PQ and QT intervals was unsatisfactory for this methodology to prevail in a clinical setting, these values provide the basis for in utero non-invasive investigation of foetal cardiac activity by magnetocardiography. PMID- 10677061 TI - Newborn infants' cry after heel-prick: analysis with sound spectrogram. AB - The aim of the study was to test the hypothesis that a newborn infant's cry can be used in conjunction with an instrument to measure pain. Crying due to pain was analysed after a heel-prick stimulus. In a prospective, descriptive study, 50 healthy newborn infants were subjected to a heel-prick for phenylketonuria screening. Their cries of pain were recorded and analysed. Duration of the crying sound was analysed and, using a sound spectrogram, the fundamental frequency and the cry melody of the first five cry sounds were analysed. The analysis showed that the crying sound after the painful stimulus of the heel-prick had a significantly higher fundamental frequency and lasted longer at the first than at the fifth cry. The first cry had a more varied crying melody than the fifth. There were large differences between individual cries from a single infant, as well as in the duration of each cry, total crying time, and fundamental frequencies between infants. While the first cry was more like a cry of pain, the fifth cry more resembled crying for reasons other than pain. The results suggest that newborn infants react to pain in a recognizable way. However, other stimuli may cause a similar reaction. Crying can therefore be used to measure pain in newborn infants only when the cause of crying is known. PMID- 10677062 TI - Intrauterine growth retardation and brainstem auditory-evoked response in preterm infants. AB - Intrauterine growth retardation is frequently associated with intrauterine undernutrition, and can deleteriously affect brain function. Twenty-eight premature small for gestational age infants were compared with 28 premature appropriate for gestational age infants to determine whether intrauterine growth retardation was associated with abnormalities in the auditory pathway in the early neonatal period. The auditory pathway was studied between 4-18 wk of life by analysis of brainstem auditory-evoked potentials elicited by a 10/s 75 decibel above normal adult hearing level (dB nHL) click stimulus presented at the infants' ears. Peak latencies of components I, III and V, and interpeak latencies I-III, III-V and I-V, yielded no statistically significant differences between groups. The present study indicates that intrauterine growth-retarded premature infants may not have abnormalities of brainstem auditory-evoked response in the early neonatal period. PMID- 10677063 TI - Unilateral posthaemorrhagic hydrocephalus in the neonatal period or later in infancy. AB - Five infants who developed unilateral hydrocephalus associated with antenatal or perinatal intraventricular haemorrhage (IVH) in the neonatal period or later in infancy are reported. Unilateral hydrocephalus occurred following discharge home in four of our five cases, two of whom had been treated during the neonatal period with either serial lumbar punctures or punctures from a Rickham reservoir. An obstruction at the level of the foramen of Monro following a large subependymal matrix bleed appeared to be the underlying aetiology. These data suggest that infants who suffer a predominantly unilateral IVH, with or without parenchymal involvement, can subsequently develop unilateral hydrocephalus. Cranial ultrasound examinations should be repeated at regular intervals during the first year of life, as unilateral hydrocephalus can still develop after a period of apparent stabilization. PMID- 10677064 TI - Neonatal micrognathia is associated with small upper airways on radiographic measurement. AB - In order to determine if infants with clinical micrognathia identified in the newborn period have smaller upper airways than do normal infants, and if their airway size is related to risk of later apnoea, respiration-timed upper airway radiographic measurements were performed in 21 asymptomatic neonates with clinical micrognathia. Their radiographic measurements were compared with those of a previously reported cohort of 35 normal infants. The micrognathic infants and a control group of 27 infants referred for parental anxiety were followed for 6 mo on home apnoea monitors. Sleep apnoea at home requiring stimulation by the parents occurred in 6 of 7 infants with micrognathia associated with craniofacial anomalies, 9 of 14 (64%) infants with isolated micrognathia, but only 1 of the 27 control infants (p < 0.001). Upper airway measurements at term of the infants with isolated micrognathia who later experienced apnoea were significantly smaller than either those of normal infants (p < 0.01) or of micrognathic infants who did not have apnoea requiring stimulation (p < 0.05). In conclusion, upper airway measurements on timed lateral radiographs in asymptomatic micrognathic infants at term (corrected age) revealed them to be smaller than those of normal infants. Narrower upper airways were associated with increased risk of subsequent apnoea requiring stimulation. PMID- 10677065 TI - Are data on the prevalence and duration of breastfeeding reliable? The case of Italy. AB - Many countries produce data on the prevalence and duration of breastfeeding, but are they reliable? We reviewed 16 studies on breastfeeding in Italy published after 1990. They report a prevalence of breastfeeding at and around birth ranging from 66% to 91%, decreasing to 17-52% at 4 mo and 28-36% at 6 mo. Most studies refer to a non-representative sample of the Italian population. Two studies used standard definitions of breastfeeding, but their results are difficult to interpret or cannot be generalized. Five other studies used non-standard definitions that undermine the interpretation of results. The remaining nine studies used no definition at all. All studies used a recall period different from 24 h, or from the whole hospital stay for breastfeeding at discharge, making the interpretation of results even more difficult. We conclude that the published information gives an inaccurate picture of the prevalence and duration of breastfeeding in Italy, leading to unjustified optimism and inaction. The actual figures may be lower, as shown by preliminary data from a small Italian region: using standard definitions and methods during a 9-mo monitoring period, exclusive breastfeeding averaged 35% at discharge and 23% at about 4 mo of age. PMID- 10677066 TI - Can severe language disability be identified in three-year-olds? Evaluation of a routine screening procedure. AB - This study evaluates the predictability of a new language screening procedure in 3-y-olds. It is used in several Child Health Centres (CHC) in Sweden and has the character of a field study involving more than 60 CHC nurses. The main questions concern the (i) development in 3-y-olds assessed as severely language delayed and (ii) whether there are any earlier unknown severely disabled children identified at 4 y of age. Ninety-six percent of the original study population participated in the follow-up. The calculations are based on results from 2237 children. A well-established screening routine, which has been shown capable of predicting the risk of not being able to follow expected schooling, and case records were used as an acceptable proxy outcome measure, pending a better gold standard. In the group of severely disabled 3-y-olds, sensitivity, specificity and positive predictive values were 86, 99 and 43%, respectively. Finally, three false negatives were identified. In light of the present results, continued application of the 3-y screening is discussed. PMID- 10677067 TI - Incidence of type I diabetes among children and young adults (0-29 years) in the province of Badajoz, Spain during 1992 to 1996. AB - In this study, we determine the incidence of Type I (insulin-dependent) diabetes mellitus in the 0-29-y-old group in Badajoz (the largest and least developed province of Spain). We test for differences in incidence by age at diagnosis, time cluster and sex. Diabetes clinics and periodic review of hospital administration data provided the primary source of ascertainment. The secondary independent data source was based on registries of local Diabetic Associations and guarantee cards of blood glucose meters. Data were collected retrospectively in the period 1992-95 and prospectively for 1996. During the 5-y period (1992 96), 186 new cases of Type I diabetes were identified. Completeness of ascertainment was 95%. Average annual incidence (95% CI) for the 0-14, 15-29 and 0-29-y-old groups was 17.6/100,000 (14.5-21.2), 8.8/100,000 (6.9-11.1) and 12.8/100,000 (11-14.7). The highest age-specific annual incidence rate was found in the 10-14 age group: 23.4/100,000 (17.6-30.4). The incidence in males (14.7/100,000/y) was higher than in females (10.7/100,000/y). There was a seasonal onset pattern, with the highest incidence in autumn and winter. October was the month with the highest number of new cases (29/186). The province of Badajoz has a moderately high incidence of Type I diabetes in 0-14-y-old children, similar to that found in other more developed and densely populated regions of Spain. These data contradict the hypothesis of a decrease in the incidence of the disease from north to south over Europe. PMID- 10677068 TI - Long-term coping in childhood cancer survivors: influence of illness, treatment and demographic background factors. AB - In 30 survivors of childhood cancer, long-term psychological coping with experience of disease and treatment was studied in relation to factors associated with illness, treatment and demographic background. Coping was assessed in a prior study, in which three groups of varying levels of coping where delineated (good, 40%; intermediate, 33%; poor, 27%, coping). The present study showed that poor individual coping was related to diagnosis, a shorter time of continuous complete remission, more severe illness and treatment impairments, and lower scores on a test of intellectual abilities. In addition, a longer time of treatment tended to be followed by poorer coping. However, no association was found for gender, parents' occupational level, age at illness onset, neuro cranial irradiation, irradiation dose (total) or age at investigation. A tentative path-analysis was executed, displaying a model for the relationships between medical and demographic background variables, and for their influence on coping. It was concluded that a complex of factors--associated particularly with severity of disease and treatment--appears to be related to, and affects, coping with the illness experience. Patients' long-term coping with their illness trauma is most likely determined by multiple factors. Intellectual capabilities are related to coping. PMID- 10677069 TI - Previous breastfeeding does not alter thymic size in infants dying of sudden infant death syndrome. AB - The relationship between thymic weights and previous feeding histories was examined in 294 infants of 37 wk gestation or more dying of sudden infant death syndrome (SIDS). One hundred and sixty-five infants had been breastfed exclusively, 89 had been partially breastfed and 40 had never been breastfed. We found no relationship between thymic weight and type of previous feeding. The difference between these findings in SIDS and the substantially greater thymic size previously reported in 4-mo-old breastfed living infants deserves further study. PMID- 10677070 TI - Blueberry muffin rash as a presentation of alveolar cell rhabdomyosarcoma in a neonate. AB - Soft tissue sarcomas of childhood continue to present problems with pathologic diagnosis, staging and treatment. Rhabdomyosarcoma, the most common soft tissue sarcoma, represents 4-8% of all malignant solid tumours in children. We report a case of congenital alveolar rhabdomyosarcoma who presented with "blueberry muffin"-like rash. A full-term female infant was noted at birth to have multiple skin lesions resembling blueberry muffin rash and an abdominal mass in the left iliac fossa, which appeared to be fixed to the posterior abdominal wall. There was no enlargement of liver and spleen, but her para-aortic lymph nodes were enlarged. Biopsy from the mass confirmed the diagnosis of alveolar cell rhabdomyosarcoma. Molecular investigation for the t (2:13) translocation was negative. The infant received chemotherapy but died within 1 mo of diagnosis. PMID- 10677071 TI - Association of oesophageal atresia and hypertrophic pyloric stenosis. PMID- 10677072 TI - The increased trend of type 1 diabetes mellitus in children (0-14 years) in the Upper Silesia region of Poland. PMID- 10677073 TI - Treatment of nocturnal enuresis. PMID- 10677074 TI - Cystic fibrosis presenting as heat stroke. PMID- 10677075 TI - Low-carbon silica sorbents for solid-phase extraction. AB - The applicability of silica gels, modified with cyclic organosiloxanes, in solid phase extraction (SPE) has been tested. Surface characteristics of the adsorbents prepared are determined by: elemental analysis, 29Si cross-polarisation magic angle spinning nuclear magnetic resonance spectroscopy (29Si CP MAS NMR) and diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. The sorbents with low carbon contents are used for extraction of chlorinated pesticides, polychlorinated biphenyls and nitro-compounds from water. The properties of the sorbents are compared with commercial C18 ones. It is shown that the low-carbon silica SPE sorbents can ensure satisfactory recoveries and reveal some selectivity. PMID- 10677076 TI - Use of optimization software to determine rugged analysis conditions in high performance liquid chromatography. AB - The ruggedness evaluation of an analytical method is now generally required for further validation. By considering ruggedness at an early stage of method development, major disappointments and amount of work could be avoided. This work shows that the optimization software OSIRIS can be helpful for the chromatographer during a method development, as it takes into account the method ruggedness. The ruggedness of the analysis conditions is then evaluated all along the selectivity optimization procedure. This optimization software belongs to the interpretive methods that consist of predicting the optimum conditions by modeling first the solute retention over the parameter space using a minimum number of preliminary runs. The choice of a response function is studied. This response function must be able to take into account several individual criteria: analysis time, minimal resolution and ruggedness of each parameter. Some optimum separations, determined using a ruggedness criteria or not, are given and compared in terms of long term repeatability. PMID- 10677077 TI - Liquid chromatographic resolution of 2-hydroxycarboxylic acids on a new chiral stationary phase derived from (S)-leucine. AB - Enantiomers of racemic 2-hydroxycarboxylic acids have been resolved as their O ethoxycarbonyl pi-basic anilide derivatives on a new chiral stationary phase (CSP) derived from N-(3,5-dinitrobenzoyl)leucine N-phenyl N-alkylamide and the resolution results have been compared with those on various commercial pi-acidic CSPs. The resolution results demonstrate that the new CSP derived from N-(3,5 dinitrobenzoyl)leucine N-phenyl N-alkylamide is most effective among the five CSPs tested for the resolution of 2-hydroxycarboxylic acid derivatives. In order to elucidate the chiral recognition mechanism exerted by the new CSP, the resolution of slightly differently modified derivatives of 2-hydroxycarboxylic acids on the new CSP has been investigated. Based on the resolution results, a chiral recognition mechanism utilizing three simultaneous interactions such as the face to face pi-pi interaction and the two hydrogen bonding interactions between the CSP and the more retained enantiomer of the analyte has been proposed. PMID- 10677078 TI - Determination of aliphatic anhydrides and acids by reversed-phase liquid chromatography. AB - Few chromatography methods have been reported for the determination of anhydrides in mixtures or as mixed anhydrides. The potential reactivity of anhydrides with water and other common eluent components complicates possible schemes for separation and analysis. By optimizing variables that affect hydrolysis, including the stationary phase, conditions can be found to successfully analyze anhydrides as reactive as acetic anhydride. The corresponding acids can be determined at the same time. The effect of the stationary phase on anhydride hydrolysis rates may prove to be a sensitive means of probing stationary phase chemistry. PMID- 10677079 TI - Does further clean-up reduce the matrix enhancement effect in gas chromatographic analysis of pesticide residues in food? AB - Sample extracts of apples, peas, green beans, oranges, raspberries, clementines, carrots, and wheat obtained using the Food and Drug Administration (acetone extraction) and Canadian Pest Management Regulatory Agency (acetonitrile extraction) multiresidue methods for pesticides were subjected to clean-up using different solid-phase extraction (SPE) cartridges in an attempt to reduce or eliminate the matrix enhancement effect. The matrix enhancement effect is related to the blocking of active sites on the injector liner by matrix components, thereby increasing signal in the presence of matrix versus standards in solvent in which the pesticides themselves interact with the active sites. Graphitized carbon black (GCB) was often used in combination with various anion-exchange SPE cartridges. The extracts were then spiked with organophosphorus insecticides. These process standards were then compared to standards in acetone of the same concentration using gas chromatography with flame photometric detection or ion trap mass spectrometric detection. Sample matrix enhancement varied from little to no effect for some pesticides (e.g. chlorpyrifos, malathion) to >200% in the case of certain susceptible pesticides. The GCB removed color components but showed little effect in reducing matrix enhancement by itself. The anion-exchange cartridges in combination with GCB or not, substantially reduced the matrix enhancement effect but did not eliminate it. PMID- 10677080 TI - Influence of pH*-value of methanolic electrolytes on electroosmotic flow in hydrophilic coated capillaries. AB - The dependency of EOF on the H+-concentration and the related so called pH* value of methanolic electrolytes has been examined with poly(ethylene glycol) (PEG), poly(vinyl alcohol) (PVA) and uncoated capillaries. These results were compared with the pH dependency of EOF of these capillaries using aqueous buffers. In uncoated capillaries the dependency of EOF on the pH(*)-value is very similar for aqueous and methanolic electrolytes. The EOF increases with increasing H+ concentration and pH-hysteresis is observed. In PVA coated capillaries the EOF is strongly reduced over wide pH* or pH ranges for both methanolic electrolytes and aqueous buffers. The EOF in PEG coated capillaries is surprisingly directed to the anode with methanolic electrolytes whereas a reduced cathodic EOF is observed in aqueous electrolytes. The anodic EOF of PEG-coated capillaries in methanolic electrolytes is independent of the pH*-value. The usefulness of PEG- and PVA coated capillaries for adjusting the EOF in non-aqueous electrolytes for the analysis of isomeric organic acids was demonstrated. PMID- 10677081 TI - Extraction of iridoid glycosides and their determination by micellar electrokinetic capillary chromatography. AB - Several methods for the extraction of two iridoid glycosides, catalpol and aucubin, from the plant matrix (Veronica longifolia leaves) were compared. Pressurized hot water extraction and hot water extraction were the most efficient isolation techniques for both. Pressurized liquid extraction and maceration with various organic solvents were also tested. Relative to the amounts extracted with hot water, ethanol extracted only 22% of catalpol and 25% of aucubin and pressurized hot water extracted 83% of catalpol and 92% of aucubin. The lowest relative standard deviations, 22% for catalpol and 8% for aucubin, were achieved with hot water extraction (13 repetitions), and the highest relative standard deviations, 76% for catalpol and 73% for aucubin, with pressurized liquid extraction (five repetitions). A fast capillary electrophoretic method was developed for the quantitative determination of catalpol and aucubin. PMID- 10677082 TI - Protein adsorption to the bare silica wall in capillary electrophoresis quantitative study on the chemical composition of the background electrolyte for minimising the phenomenon. AB - A novel method is reported for quantifying protein adsorption to naked silica tubings and for assessing the efficacy of amino quenchers added to the background electrolyte. It consists of flushing a fluorescently-labelled protein (myoglobin) into a capillary equilibrated in Tris-acetate buffer, pH 5.0, until full saturation of the potential adsorbing sites. Desorption is then affected by driving electrophoretically sodium dodecyl sulphate (SDS) micelles into the capillary from the cathodic reservoir: the peak of eluted material is quantified fluorometrically by using a dual laser beam instrument able to read the fluorescein-isothiocyanate-labelled myoglobin at 520 nm and the internal standard (sulphorodamine) at 630 nm. As potential quenchers, a series of monoamines have been investigated (triethylamine, triethanolamine, ethylamine), followed by diamines (putrescine, cadaverine and hexamethonium bromide) and finally by oligoamines [spermidine, spermine and TEPA (tetraethylenepentamine), i.e., a tri- a tetra- and a pentamine, respectively]. Two values of molarities have been derived: a value at 50% (a kind of a dissociation constant) and a value at 90% inhibition of binding of macromolecules to the silica surface. According to these figures of merit, mono- and diamines are rather poor quenchers of interaction with the wall, since the 50% values are of the order of 50-100 mM and the 90% values reach as high as 560 mM. On the contrary, oligoamines, especially spermine and TEPA, are most effective, since the 50% molarities are in the sub-millimolar range and the 90% values are of the order of ca. 1 mM. Figures of merit have also been derived for different washing procedures. Those most commonly adopted in routine practice, i.e., of washing with either 1 M NaOH or with 1 M HCl, or with both, leave behind traces of proteins still bound to the wall, whereas the SDS micelle electrophoretic desorption seems to be 100% effective. PMID- 10677083 TI - Determination of diclofenac sodium by capillary zone electrophoresis with electrochemical detection. AB - Capillary zone electrophoresis was employed for the determination of diclofenac sodium using an end-column amperometric detection with a carbon fiber microelectrode, at a constant potential of 0.83 V vs. saturated calomel electrode. The optimum conditions of separation and detection are 4.90 x 10(-3) mol/l Na2HPO4-3.10 x 10(-3) mol/l NaH2PO4 (pH 7.0) for the buffer solution, 10 kV for the separation voltage, 5 kV and 10 s for the injection voltage and the injection time, respectively. The limit of detection is 2.5 x 10(-6) mol/l or 5.2 fmol (S/N=2). The relative standard deviation is 0.8% for the migration time and 4.7% for the electrophoretic peak current. The method was applied to the determination of diclofenac sodium in human urine. PMID- 10677084 TI - Use of high-performance liquid chromatographic fractionation of large RNA molecules in the assay of group I intron ribozyme activity. AB - Ion-pair reversed-phase high-performance liquid chromatography (HPLC), which has previously been used to fractionate double-stranded DNA molecules, can be applied to single-stranded RNA molecules in the size range of 200-1000 nucleotides. This procedure permits RNA molecules to be separated and recovered rapidly in liquid medium, thereby facilitating recovery. We have used this system to separate an in vitro transcription product containing a group I intron ribozyme from the intermediates and products of the splicing reaction, permitting rapid assay of ribozyme activity without the use of radioactivity. PMID- 10677085 TI - Determination of interfering triazine degradation products by gas chromatography ion trap mass spectrometry. AB - Deethylatrazine (DEA), an atrazine degradation product, has been added to the US Environmental Protection Agency's Drinking Water Contaminant Candidate List (CCL). In its gas chromatographic analysis, DEA can coelute with deisopropylatrazine (DIA), another degradation product. The present work demonstrates that the coelution of DEA and DIA can induce a significant (up to approximately 50%) positive bias in the DEA determination, when using an ion-trap mass spectrometer as the detector. The DIA determination is unaffected by the coelution within experimental error. This may be explained in terms of gas-phase ion fragment populations. A correction factor to the observed DEA concentration may be developed based on the measured DIA concentration. PMID- 10677086 TI - Apparatus and method for interfacing capillary electrophoresis and chemiluminescence nitrogen detection for the analysis of nitrogen-containing compounds. AB - We have developed an interface that allows the specific detection of nitrogen containing compounds by using a chemiluminescence nitrogen detector. The feasibility of using this interface was demonstrated by separating and detecting two nitrogen-containing compounds, p-aminosalicylic acid and L-phenylalanine. Although baseline separation was achieved, the theoretical plates were lower when compared to UV detection (25000 vs. approximately 85000). A sensitivity of 75 ng (approximately 500 pmol) per injection was achieved with this system which is adequate for pharmaceutical and biotech applications. PMID- 10677087 TI - Determination of peroxides by capillary zone electrophoresis with amperometric detection. AB - The combination of cathodic amperometric detection with capillary zone electrophoresis is demonstrated to be a versatile method for the quantification of organic and inorganic peroxides. A gold microelectrode, polarized at -600 mV against an Ag/AgCl reference electrode, is placed at the end of the capillary. Since the electroosmotic flow purges the detector electrode from oxygen, no degassing of the detector cell or the sample is necessary. With an injection volume of ca. 1 nl, hydrogen peroxide, peroxosulfate, peroxy alkanoic acids and the hydroperoxides of linoleic acid can be detected down to 10 micromol/l. Separation of the isomeric hydroperoxides of the unsaturated fatty acids is achieved by addition of beta-cyclodextrin to the electrolyte. PMID- 10677088 TI - Ion exchange-based preconcentration for the determination of anions by capillary electrophoresis. AB - In the present paper a capillary zone electrophoresis (CZE)-compatible preconcentration technique for anions, based on ion exchange, is described. The described preconcentration approach has found limited use until recently because of the inherent elution step that leads to contamination of the sample with eluent components. In this paper, we describe an improved anion exchange-based preconcentration technique in which contamination of the sample with the eluent constituents, which occurs during anion elution from the preconcentration column, is eliminated by on-line chemical suppression on a packed-bed suppressor column. In the present communication, the basic principles of the proposed anion enrichment system are presented. The system was optimized, resulting in a minimal additional dilution of the eluted sample plug. This was achieved by the use of a computer-controlled, sensing/switching system. The effectiveness of the developed method was later tested on the determination of some anions in a synthetic sample using CE apparatus. PMID- 10677089 TI - Chromatographic analysis of bisphosphonates. PMID- 10677090 TI - Low-dose methotrexate controls a severe form of polyarteritis nodosa. PMID- 10677091 TI - "Strawberry" gingival hyperplasia: a pathognomonic mucocutaneous finding in Wegener granulomatosis. PMID- 10677092 TI - Serum levels of autoantibodies to BP180 correlate with disease activity in patients with bullous pemphigoid. AB - OBJECTIVE: To investigate the possible correlation of levels of circulating anti BP180 autoantibodies with disease activity in bullous pemphigoid (BP). DESIGN: Diagnostic study. SETTING: Regional referral center at a university dermatology department. PATIENTS: Fifteen patients with typical clinical, histologic, and immunofluorescence findings of BP who had not received prior systemic treatment. INTERVENTIONS: Initially, 6 consecutive patients with BP were treated with oral doxycycline and niacinamide. Subsequently, 9 consecutive patients with BP received a combination of oral dapsone and prednisolone. MAIN OUTCOME MEASURES: Disease activity, serum levels of autoantibodies to BP180, and titers of antibasement membrane zone autoantibodies were assayed before initiation of treatment and 4 and 8 weeks later. Reactivity to BP180 was analyzed by enzyme linked immunosorbent assay using a recombinant form of BP180 NC16A. Titers of anti-basement membrane zone autoantibodies were assayed by indirect immunofluorescence on 1-mol/L sodium chloride-split human skin. RESULTS: In both treatment groups, disease activity correlated with serum levels of autoantibodies to BP180 NC16A (P = .004 [dapsone-prednisolone] and .007 [doxycycline niacinamide]). No correlation was seen between disease activity and indirect immunofluorescence reactivity (P = .18 and .16, respectively). In patients receiving dapsone plus prednisolone, the dose of corticosteroids necessary to suppress new blister formation correlated with anti-BP180 reactivity (P = .002). CONCLUSIONS: In contrast to indirect immunofluorescence reactivity that reflects reactivity to both BP 180 and BP230, serum levels of autoantibodies to BP180 correlate with disease activity in BP. Assaying reactivity to BP180 should be a helpful guide for the therapeutic management of patients with this disease. Our results underline the pathogenic relevance of autoantibodies to human BP180. PMID- 10677093 TI - A randomized, 12-year primary-prevention trial of beta carotene supplementation for nonmelanoma skin cancer in the physician's health study. AB - CONTEXT: Although basic research provides plausible mechanisms for benefits of beta carotene supplementation on nonmelanoma skin cancer (NMSC) primarily consisting of basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), observational studies are inconsistent. Randomized trial data are limited to 1 trial of secondary prevention that showed no effect of beta carotene on the incidence of NMSC after 5 years. OBJECTIVE: To test whether supplementation with beta carotene reduces the risk for development of a first NMSC, including BCC and SCC. DESIGN: Randomized, double-blind, placebo-controlled trial with 12 years of beta carotene supplementation and follow-up. SETTING: Physicians' Health Study in the United States. PARTICIPANTS: Apparently healthy male physicians aged 40 to 84 years in 1982 (N = 22 071). INTERVENTION: Beta carotene, 50 mg, on alternate days. MAIN OUTCOME MEASURE: Relative risk (RR) and 95% confidence interval (CI) for a first NMSC, BCC, and SCC. RESULTS: After adjusting for age and randomized aspirin assignment, there was no effect of beta carotene on the incidence of a first NMSC (RR, 0.98; 95% CI, 0.92-1.05), BCC (RR, 0.99; 95% CI, 0.92-1.06), or SCC (RR, 0.97; 95% CI, 0.84-1.13). There was also no significant evidence of beneficial or harmful effects of beta carotene on NMSC by smoking status (current, past, or never). CONCLUSION: This large-scale, randomized, primary prevention trial among apparently healthy well-nourished men indicates that an average of 12 years of supplementation with beta carotene does not affect the development of a first NMSC, including BCC and SCC. PMID- 10677094 TI - Decision support software to help primary care physicians triage skin cancer: a pilot study. AB - OBJECTIVE: To determine whether decision support software can help primary care physicians proficiently triage lesions suggestive of basal cell and squamous cell carcinoma. DESIGN/MEASURES: Physicians selected triage options for 15 digitized images of skin lesions, with and without use of the decision support software. PARTICIPANTS/SETTINGS: Twenty primary care physicians practicing in a health maintenance organization or a city health clinic. INTERVENTION: Decision support software designed to help physicians arrive at a triage recommendation consisted of a clinical information form, a decision tree, and support features (teaching points, example images, and diagrams). RESULTS: Without using the decision support software, physicians chose the wrong triage decision 36.7% of the time; using the decision support software, they chose the wrong response only 13.3% of the time. Not using the decision support software, they failed to correctly perform a biopsy on or refer patients with cancerous lesions 22.1% of the time; using the software, they failed to correctly perform a biopsy on or refer patients with cancerous lesions only 3.6% of the time. Physicians scored an average of 3 points (of a possible 15 points) higher when they used the software (signed rank, 101.0; P<.001). They scored an average of 1 point higher on the 7 cancerous lesions when they used the software (signed rank, 65.5; P<.001). CONCLUSIONS: Use of decision support software could improve primary care physicians' triage decisions for lesions suggestive of nonmelanoma skin cancer, and potentially reduce morbidity and health care costs. We are designing a larger study to evaluate the accuracy and utility of the software with patients seen in clinical practice. PMID- 10677095 TI - Expression of p53 in arsenic-related and sporadic basal cell carcinoma. AB - BACKGROUND: The TP53 gene has been shown to have an important role in the genesis of sporadic, presumably mainly sunlight-related, basal cell carcinoma (BCC). However, its role in arsenic-related BCCs is not clear, although the trivalent form of arsenic has been long recognized as a cause of BCC. Arsenic treatment has been shown to cause hypermethylation of the TP53 gene in lung carcinoma cell lines, but it is not known if this occurs in vivo in arsenic-related BCCs. OBJECTIVE: To compare the immunohistochemical expression of the p53 protein in arsenic-related and sporadic BCCs to determine if the expression pattern is consistent with gene silencing. SETTING: A research institute and hospital in Australia. CASES: One hundred seventeen white patients with 121 sporadic BCCs and 21 white patients with 92 arsenic-related BCCs. MAIN OUTCOME MEASURES: The expression and the intensity of p53 were scored semiquantitatively. Statistical analysis was performed using the chi2 test. RESULTS: Arsenic-related BCCs express p53 less often and at a lower intensity than sporadic BCCs (P = .001; 2-tailed test). The BCCs from sun-exposed sites, whether arsenic related or sporadic, more frequently showed overexpression of p53 than those from less-exposed areas (P = .004; 2-tailed test). The more aggressive subtypes of BCC show a higher level of expression of p53 than the less aggressive forms (P = .04; 2-tailed chi2 test). CONCLUSIONS: These results are consistent with the hypothesis that the TP53 gene is down-regulated by methylation in arsenic-related BCC, particularly those from less-exposed sites. However, an alternative possibility is that mutations in TP53 that stabilize the protein are less common in arsenic-related BCCs. Further analysis will be necessary to distinguish between these hypotheses. PMID- 10677096 TI - Melanoma screening: report of a survey in occupational medicine. AB - BACKGROUND: Epidemiological studies concerning melanoma are most often performed by general practitioners and dermatologists in patients previously aware of the risk of nevi. OBJECTIVE: To determine the efficiency of early detection of melanoma by occupational medicine specialists trained in the use of ABCDE criteria during annual systematic examination of workers. METHODS: A total of 370 subjects with suspect lesions that demonstrated at least 2 of 5 ABCDE criteria were selected from 65000 employees examined; these subjects were requested to see their physician about possible excision. Of the 370 subjects, 273 (73.8%) were seen at a second-year follow-up visit to determine their outcome. RESULTS: Among the 273 subjects who were seen again, 172 (63.0%) had consulted a physician. For the 101 subjects who had not seen a physician, the main reason was the negligence (86.1%). A total of 353 atypical nevi were observed. The mean number of ABCDE criteria noted per lesion was 2.6. Lesion diameter greater than 6 mm was the most frequent (80.5%) and enlargement the least frequent criteria seen; heterochromous coloration and diameter greater than 6 mm was the most common association (54.5%). Five histologically confirmed melanomas were found among nevi excised in 78 subjects. CONCLUSION: This screening approach seems efficient for the early detection of melanoma, demonstrating an incidence of 7.7 per 100000 vs. 9 per 100000 in the general French population. PMID- 10677097 TI - Fibrosing alopecia in a pattern distribution: patterned lichen planopilaris or androgenetic alopecia with a lichenoid tissue reaction pattern? AB - BACKGROUND: Androgenetic alopecia is characterized by a defined area of progressive nonscarring alopecia. The clinical and histological findings in 15 women and 4 men with progressive scarring alopecia in a pattern distribution were studied. The results were evaluated and compared with clinicopathologic entities that feature scarring of the central scalp area, specifically, lichen planopilaris, pseudopelade, and follicular degeneration syndrome. OBSERVATIONS: Patients developed progressive fibrosing alopecia of the central scalp, without the multifocal areas of involvement typical of lichen planopilaris and pseudopelade. Perifollicular erythema, follicular keratosis, and loss of follicular orifices were limited to a patterned area of involvement. Biopsy specimens of early lesions demonstrated hair follicle miniaturization and a lichenoid inflammatory infiltrate targeting the upper follicle region. Advanced lesions showed perifollicular lamellar fibrosis and completely fibrosed follicular tracts indistinguishable from end-stage lichen planopilaris, pseudopelade, or follicular degeneration syndrome. CONCLUSIONS: Some patients with androgenetic alopecia might have additional clinical and histological features of inflammation and fibrosis limited to the area of androgenetic hair loss. In these patients, the histological findings of early lesions are identical to those seen in lichen planopilaris. The lichenoid tissue reaction leading to follicular destruction in these patients might be pathogenetically related to the events underlying androgenetic alopecia. PMID- 10677098 TI - Clinical findings in mosaic carriers of hypohidrotic ectodermal dysplasia. AB - BACKGROUND: Hypohidrotic ectodermal dysplasia (HED) is a severe developmental disorder in which nonallelic genetic heterogeneity has been demonstrated. Even though X-linked and autosomal recessive forms are phenotypically similar, identification of the way of transmission is mandatory to give reliable genetic counseling to the family and to address molecular studies. Complete examination of relatives of patients with HED and identification of carriers of partial forms of the disorder in their families is the key to clarifying intrafamilial genetic transmission. OBSERVATIONS: Seven patients diagnosed as having HED and their first-degree relatives were carefully examined and tested with starch-iodine. Useful signs for identifying possible carriers of and postzygotic mosaics for X linked HED and for finding distinctive features between the X-linked and the autosomal recessive forms of the disorder were recorded. Of these, the most striking finding was the clinical evidence of the distribution of normal and abnormal skin along Blaschko lines in heterozygous and postzygotic mutation carriers of X-linked HED. Six heterozygous female carriers of X-linked HED, 2 males with postzygotic mutations for X-linked HED, and 1 female with autosomal recessive HED were clinically identified. At the end, 6 families had a diagnosis of X-linked HED, while 1 had a diagnosis ofautosomal recessive HED. Clinical data, family history, and starch-iodine test results were never in conflict in the 7 families. CONCLUSIONS: Careful clinical examination is the best way to detect heterozygous carriers and postzygotic mutation of X-linked HED. Heterozygous parents of patients with autosomal recessive HED show no features of the disorder. The starch-iodine test is not superior to a clinical examination in heterozygous carrier detection but may play a confirmative role and be of help in differentiating X-linked and autosomal recessive HED in isolated patients. PMID- 10677099 TI - Inflammatory variant of epidermolysis bullosa acquisita with IgG autoantibodies against type VII collagen and laminin alpha3. AB - BACKGROUND: The inflammatory variant of epidermolysis bullosa acquisita (EBA) may clinically closely resemble bullous or cicatricial pemphigoid. Patients with inflammatory EBA have IgG autoantibodies against type VII collagen. Patients with anti-epiligrin cicatricial pemphigoid have IgG autoantibodies against laminin 5. OBSERVATION: We describe a patient with inflammatory EBA exhibiting nonscarring oral and vaginal involvement. Indirect immunofluorescence using skin substrate lacking an epidermal basement membrane molecule, direct immunoelectron microscopy, immunoblot, and immunoprecipitation studies revealed the simultaneous presence of circulating IgG autoantibodies against type VII collagen and laminin alpha3. A final diagnosis of EBA was based on the sublamina densa level of blister formation. CONCLUSION: This case illustrates the clinical and immunological overlap between EBA and anti-epiligrin cicatricial pemphigoid, a unique finding that may have developed as a consequence of epitope spreading. PMID- 10677100 TI - A new look at scarring alopecia. PMID- 10677101 TI - Prevention of nonmelanoma skin cancer. PMID- 10677102 TI - Decision support is changing health care. PMID- 10677103 TI - Levels of antibodies to BP180 correlate with disease activity in bullous pemphigoid. PMID- 10677104 TI - Recalcitrant patch of alopecia on the scalp. PMID- 10677105 TI - Acquired nodule on the right side of the nose. PMID- 10677106 TI - Truncal hypopigmented macules and facial hyperpigmented papules. PMID- 10677107 TI - Penile necrosis. PMID- 10677108 TI - "Significant" scientific productivity should be weighed against the expenses necessary to finance it. PMID- 10677109 TI - Use caution when establishing "routine" prophylactic antibiotic procedures. PMID- 10677110 TI - The ethical issues of the Holmesburg studies have been addressed. PMID- 10677111 TI - Ethical accusations: the loss of common sense. PMID- 10677112 TI - Scarring following Q-switched laser treatment of "double tattoos". PMID- 10677113 TI - Blood transfusions and psoriasis: is there a link? PMID- 10677114 TI - Systemic photodynamic therapy is a safe and effective treatment for psoriasis. PMID- 10677115 TI - North American Contact Dermatitis Group patch-test results, 1996-1998. PMID- 10677116 TI - The prevalence and incidence of atopic dermatitis in a birth cohort: the importance of a family history of atopy. PMID- 10677117 TI - Acrosclerosis in patients with systemic sclerosis responds to low-dose UV-A1 phototherapy. PMID- 10677118 TI - Blood screening--the next generation in testing. PMID- 10677119 TI - Use of cytokeratin fragments 19.1 and 19.21 (Cyfra 21-1) in the differentiation of malignant and benign pleural effusions. AB - BACKGROUND: Differentiation between malignant and benign pleural effusions is often difficult. Serum level of Cyfra 21-1, a marker of cytokeratin 19 fragments, has been used in the diagnosis and monitoring of epithelial tumours, especially bronchogenic carcinomas. AIM: This study is designed to establish the usefulness of effusion Cyfra 21-1 level in differentiating malignant from benign effusions. METHODS: Forty-eight malignant effusion aspirates (proven by cytology or pleural biopsy) and 34 benign samples were compared. Cyfra 21-1 concentration was measured by a solid phase sandwich radioimmunoassay (Centocur, USA). RESULTS: Cyfra 21-1 level was significantly higher in malignant effusions (geometric mean 123.6 ng/mL, 95% confidence interval [CI] 76.6-199.4) than in benign ones (geometric mean 14.3 ng/mL, 95% CI 8.5-23.9), p<0.00005. By Receiver Operating Characteristics curve analysis, the sensitivity is 77% for a specificity of 79% if the cut-off is set at 32 ng/mL. No significant difference was observed (p=0.1) in Cyfra 21-1 concentration between adenocarcinoma and mesothelioma effusions. Cyfra 21-1 level was not influenced by the effusion protein concentration (r=0.29), or by renal function as measured by serum creatinine (r=0.1). There was no significant difference between Cyfra 21-1 levels in benign exudate and transudate effusions, p=0.28. CONCLUSIONS: Cyfra 21-1 is a useful adjunct in the workup of effusions but should not replace conventional investigations as there is considerable overlap in levels between benign and malignant groups. It is unable to differentiate between subgroups of malignancies. PMID- 10677120 TI - Renal sarcoidosis in Christchurch, New Zealand 1970-1998. AB - AIM: To identify patients presenting to a nephrologist in whom a diagnosis of sarcoidosis could be made, to assess the relevant causes of renal involvement and to review treatment and long-term follow-up of this group. METHOD: A retrospective review of the computer database PROTON for patients given the diagnosis of sarcoidosis, followed by a case note review of identified patients with respect to the mode of presentation, clinical and laboratory features, treatment and subsequent follow-up. RESULTS: Nineteen patients (15 males) were identified, mean age 45 years, all were Caucasian, and follow-up was four months to 26 years (mean 9.3 years). Most common mode of presentation was acute renal failure (11) during spring/summer (14). Evidence for systemic disease was present in all patients. Mean plasma creatinine on presentation was 0.52 mmol/L and calcium 3.01 mmol/L. Hypercalcaemia was present in 60%. Kidney biopsy was performed in seven patients with the predominant findings of tubular atrophy and interstitial fibrosis; significant granulomata were present in only two. Treatment in all patients was with corticosteroids with good result. Mean long term plasma creatinine was 0.17 mmol/L at 9.3 years. Steroid withdrawal was attempted in all patients, successful in five, with the mean time to relapse of five months in the remaining 14. Mean steroid dose in this group was 7.6 mg on long term follow-up. CONCLUSIONS: Sarcoidosis causes renal dysfunction mainly through altered calcium metabolism. Treatment with corticosteroids is successful in improving renal function, but relapse is common on steroid withdrawal and prolonged treatment is necessary for disease control. PMID- 10677121 TI - Severity of liver disease in hepatitis C infection contracted through injecting drug use. AB - BACKGROUND: Injecting drug use (IDU) is currently the most common route of hepatitis C virus (HCV) transmission in Australia and many other Western countries. Most reports on the natural history of HCV have examined populations that included patients from all risk groups, but it is possible that this increasingly important subgroup is different. AIMS: To assess the severity of liver disease in individuals who acquired HCV through IDU. METHODS: Three hundred and forty-six patients with confirmed HCV infection and a history of IDU, who had had a liver biopsy performed were recruited from a liver clinic. Demographic data, liver function tests and hepatitis B serology were obtained on all patients. A detailed drug use history and HCV viral studies were also available in a subgroup of 142 patients. RESULTS: Mean age of the group was 34 years and 73% were male. Mean duration of HCV infection was 14.6 years. Forty one per cent were infected with genotype 3a, 19% - 1a, 17% - 1 (nonsubtypable), 14% - 1b and 4% - 2b. Cirrhosis was present in 12% of patients. Patients with cirrhosis (38 years) were older than those with chronic hepatitis (34 years; p=0.0003) and had a longer duration of infection (17.2 vs 14.3 years; p=0.003). On multivariate analysis, however, patient age was the only factor independently associated with cirrhosis (odds ratio 4.2; 95% confidence interval 1.4-12.6). CONCLUSION: While cirrhosis is less common in this group than in other HCV infected populations, its prevalence may increase as these patients are followed over a longer period of time. PMID- 10677122 TI - Cardiac surgery in octogenarians--The Green Lane Hospital Experience 1995-1998. AB - BACKGROUND: An increasing number of patients aged 80 years and over are being considered and accepted for cardiac surgery. AIM: To review the experience of surgery in this elderly group of patients at our institution. METHODS: Hospital records of octogenarians undergoing surgery between January 1995 and September 1998 were reviewed and follow-up was obtained by general practitioner (GP) and patient questionnaires. RESULTS: Thirty-seven patients underwent cardiac surgery. The mean age was 82.8+/-1.4 years (range 80.8 to 86.2 years). Twenty-three (62%) were male. All were independent pre-operatively with severe symptoms and minor co existing morbidity. All operations were urgent except two (emergency). Twenty patients (54%) had isolated coronary surgery, six (16%) aortic valve replacement alone, and 11 (30%) combined surgery. There were four (11%) early deaths and five (14%) peri-operative neurological events. The mean duration of post-operative intensive care stay was 2.4+/-3.9 days (range 0.05 to 16, median 1.0) and post operative hospital stay 14.0+/-13.9 days (range 0 to 79, median 11). At the time of follow-up (mean duration 20.0+/-11.2 months) two further patients had died (non-cardiac). Twenty-six of the 31 survivors were living at home (23 independently), one with relatives, and four in residential care. Their cardiac symptoms were well controlled. The GPs of all hospital survivors, and all surviving patients themselves, felt that cardiac surgery had been beneficial. CONCLUSIONS: Cardiac surgery in the very elderly has been reserved for those with severe disease or symptoms and little co-morbidity. Early mortality is higher than for the general population undergoing cardiac surgery, but post-operative resource use is acceptable and the intermediate-term outcome for survivors is good. PMID- 10677123 TI - Evaluation of antibodies to beta2-glycoprotein 1 in the causation of coronary atherosclerosis as part of the antiphospholipid syndrome. AB - BACKGROUND: Anticardiolipin antibodies (aCL) are associated with accelerated coronary atherosclerosis. Beta2-glycoprotein 1 is a cofactor necessary for the binding of aCL. AIM: The aim of this study was to determine whether antibodies to beta2-glycoprotein 1 (anti-beta2GP1) predispose to coronary artery disease (CAD), and whether the measurement of anti-beta2GP1 will be more useful than aCL alone in the evaluation of coronary risk. METHODS: Persons who had undergone coronary angiography were invited to participate, and risk factors for coronary atherosclerosis recorded. IgG aCL and anti-beta2GP1 were measured and fasting triglyceride (TG) and total cholesterol (TC) levels were determined. Angiographic score (AS) was defined as the number of diseased vessels (0, 1, 2, 3), (>50% stenosis). Ethics Committee approval was obtained. Statistical comparison used the Student's t test and Chi-squared test. RESULTS: Ninety-seven subjects (63 male) with age range 38-81 years (mean 66.0) participated. There were 31 subjects with AS=0, 27 with AS=1, 22 with AS=2, and 17 with AS=3. The three subjects with positive aCL all had CAD, as did three of the four subjects with positive anti beta2GP1. Among patients with CAD, there was an equal incidence (4.5%, three/66) of aCL and anti-beta2GP1, and an incidence of either aCL or anti-beta2GP1 of 7.6% (five/66). Compared to the group with AS=0, those with AS=1, 2 or 3 comprised a higher mean age (p=0.001) however, there was no significant difference in the prevalence of other coronary risk factors between the two groups. There was no difference in the proportions of patients with either aCL or anti-beta2GP1 in the group with AS=1, 2, or 3, compared to the group with AS=0 (5/66 c.f. 1/31, chi2=0.146, p>0.5). CONCLUSIONS: Our study has not supported an association between anti-beta2GP1 and CAD. The measurement of anti-beta2GP1 (or aCL) in the investigation of premature CAD is not justified on the basis of our results. PMID- 10677124 TI - The effect of parental smoking on presence of wheez or airway hyper responsiveness in New South Wales school children. AB - BACKGROUND AND AIMS: To assess accurately the effect of parental smoking on the respiratory health of New South Wales (NSW) school children, we obtained a large data set by pooling data from seven cross-sectional studies conducted in NSW between 1991 and 1993. METHODS: A random sample of 6394 children age eight to 11 years was studied. Respiratory symptoms, family history of asthma and parental smoking history were measured by questionnaire, atopy by skin prick test and airway hyper-responsiveness (AHR) by histamine inhalation test. RESULTS: In total, 58.3% of children had at least one parent who smoked; 38.5% were exposed to maternal smoking. After adjusting for potential confounders, such as atopy, parental history of asthma and bronchitis in the first two years, children who were exposed to maternal smoking had a significantly increased risk of recent wheeze but not of AHR (odds ratios 1.33; 95% CI: 1.2-1.5 and 1.00; 95% CI: 0.9 1.2). CONCLUSIONS: The positive association with wheeze and the lack of an association with AHR suggests that exposure to parental smoking leads to wheezing, but does not increase airway responsiveness. PMID- 10677125 TI - New treatments for multiple sclerosis. PMID- 10677126 TI - Bedside pacetermination of arrhythmias using an explanted automatic defibrillator. PMID- 10677127 TI - Systematic reviews and geriatric interventions--the need for scientific methodology. PMID- 10677128 TI - Medicine at the beginning of the 21st century: a personal viewpoint--a peripatetic overview. PMID- 10677129 TI - Extracranial vertebral artery dissection presenting as neurogenic pulmonary oedema. PMID- 10677130 TI - Asp or spa (snakebite or drowning)? PMID- 10677131 TI - Oestrogen and vascular disease. PMID- 10677132 TI - Adult onset restricted fish allergy. PMID- 10677133 TI - Gemcitabine and the blood brain barrier. PMID- 10677134 TI - Multiple organ failure related to pantoprazole. PMID- 10677135 TI - Irukandji-like syndrome in Victoria. PMID- 10677136 TI - Faecal occult blood test screening for colorectal cancer. PMID- 10677137 TI - Trepopnoea due to positional narrowing of the left main bronchus. PMID- 10677138 TI - Surgical palliation of carcinoid syndrome. PMID- 10677139 TI - Post obstructive pneumonia secondary to endobronchial tuberculosis--an institutional review. PMID- 10677140 TI - Pulmonary and meningeal cryptococcosis in pulmonary alveolar proteinosis. PMID- 10677141 TI - Health complaints related to pesticide stored at a public health clinic. AB - Employees at a health center in Georgia were concerned that symptoms experienced by some employees were related to pesticide exposure at the center. Malathion and DDT, used for mosquito control from 1969 to 1981, had been stored and handled at the center's first floor. We surveyed 117 (91%) of 129 employees to determine whether reported symptoms were associated with pesticide exposure. We performed environmental sampling for pesticides. We analyzed serum samples for 17 chlorinated pesticides, and urine samples for malathion. We found that 37% of the participants had reported a diagnosis of sinusitis and 24% of bronchitis since working at the health center. Frequently reported symptoms were eye irritation (44%) and headache (68%). DDT and malathion were found at levels of 2.4 and 11%, respectively, in bulk samples from the loading dock of the building. Multivariate analysis of responses to the questionnaire showed that the perception of odors, inadequate air flow, and length of employment were significantly associated with the employees' health complaints. Pesticide concentrations in employees' serum and urine samples were not associated with any health complaint. The health complaints reported by the employees at the health center were precipitated by both environmental and psychological factors. The epidemiology and laboratory components of this study highlight the importance of obtaining biological measurements in episodes of perceived environmental exposure. PMID- 10677142 TI - Effects of taurine on ozone-induced memory deficits and lipid peroxidation levels in brains of young, mature, and old rats. AB - To determine the antioxidant effects of taurine on changes in memory and lipid peroxidation levels in brain caused by exposure to ozone, we carried out two experiments. In the first experiment, 150 rats were separated into three experimental blocks (young, mature, and old) with five groups each and received one of the following treatments: control, taurine, ozone, taurine before ozone, and taurine after ozone. Ozone exposure was 0.7-0.8 ppm for 4 h and taurine was administered ip at 43 mg/kg, after or before ozone exposure. Subsequently, rats were tested in passive avoidance conditioning. In the second experiment, samples from frontal cortex, hippocampus, striatum, and cerebellum were obtained from 60 rats (young and old), using the same treatments with 1 ppm ozone. Results show both an impairment in short-term and long-term memory with ozone and an improvement with taurine after ozone exposure, depending on age. In contrast to young rats, old rats showed peroxidation in all control groups and an improvement in memory with taurine. When taurine was applied before ozone, we found high peroxidation levels in the frontal cortex of old rats and the hippocampus of young rats; in the striatum, peroxidation caused by ozone was blocked when taurine was applied either before or after ozone exposure. PMID- 10677143 TI - Effects of Great Lakes fish consumption on brain PCB pattern, concentration, and progressive-ratio performance. AB - This study investigated the effects of consumption of Great Lakes fish on progressive ratio performance, and on the pattern and concentrations of brain polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethene (DDE), and mirex in the rat. Adult, male Sprague-Dawley rats were fed a 30% diet of either Lake Ontario salmon (LAKE), Pacific Ocean salmon, or lab chow control for 20 or 65 days. Following the treatment regimen, half the rats from each group were sacrificed immediately for gas chromatographic analysis of organochlorine contaminants, and the other half were tested on a multiple fixed-ratio progressive-ratio reinforcement schedule and then sacrificed for analysis. Consumption of Lake Ontario fish resulted in significantly higher levels of brain PCBs, DDE, and mirex relative to controls, but still well within human exposure ranges (<1 microg/g fat). Consumption of Lake Ontario fish for 20 or 65 days produced an average brain PCB concentration of 457 and 934 ng/g fat, respectively. Consumption of laboratory rat chow or Pacific Ocean salmon for 20 or 65 days produced an average brain PCB concentration of 240, 464, and 441 ng/g fat, respectively. Moreover, both LAKE-fed groups showed a much more heavily chlorinated pattern of brain PCBs than all control groups, as evidenced by both significant increases in the most heavily chlorinated PCB congeners and significant increases in the average chlorine biphenyl. All LAKE brains contained significant concentrations of DDE and mirex, whereas no control brains contained any detectable quantities. Analysis of progressive-ratio performance indicated that LAKE rats responded normally during fixed-ratio schedules but quit significantly sooner than control rats on a progressive-ratio 5 (PR5) schedule, indicating reduced persistence on progressively leaner reinforcement schedules. Analysis of brain PCBs indicated that total PCBs were most strongly related to PR5 performance. These data indicate that consumption by rats of contaminated Lake Ontario fish produces (1) increased concentrations of PCBs, DDE, and mirex in the brain, (2) a more heavily chlorinated distribution of PCBs in the brain, and (3) reduced persistence of progressive-ratio reinforcement schedules. While these behavioral changes are related to brain PCB level, more work is necessary before the effects can be directly attributed to PCBs. PMID- 10677144 TI - Spatially resolved hazard and exposure assessments: an example of lead in soil at Lavrion, Greece. AB - Spatially resolved hazard assessment (SRHA) and spatially resolved exposure assessment (SREA) are methodologies that have been devised for assessing child exposure to soil containing environmental pollutants. These are based on either a quantitative or a semiquantitative approach. The feasibility of the methodologies has been demonstrated in a study assessing child exposure to Pb accessible in soil at the town of Lavrion in Greece. Using a quantitative approach, both measured and kriged concentrations of Pb in soil are compared with an "established" statutory threshold value. The probabilistic approach gives a refined classification of the contaminated land, since it takes into consideration the uncertainty in both the actual measurement and estimated kriged values. Two exposure assessment models (i.e., IEUBK and HESP) are used as the basis of the quantitative SREA methodologies. The significant correlation between the blood-Pb predictions, using the IEUBK model, and measured concentrations provides a partial validation of the method, because it allows for the uncertainty in the measurements and the lack of some site-specific measurements. The semiquantitative applications of SRHA and SREA incorporate both qualitative information (e.g., land use and dustiness of waste) and quantitative information (e.g., distance from wastes and distance from industry). The significant correlation between the results of these assessments and the measured blood-Pb levels confirms the robust nature of this approach. Successful application of these methodologies could reduce the cost of the assessment and allow areas to be prioritized for further investigation, remediation, or risk management. PMID- 10677145 TI - Determination of spatial continuity of soil lead levels in an urban residential neighborhood. AB - This study uses geostatistical techniques to model and estimate soil lead levels in an urban, residential neighborhood. Sixty-two composite soil samples (median 1773 ppm; range 175 to 7953 ppm) in a four-block area of brick and stone homes were obtained. The spatial continuity of soil lead levels was modeled with a semi variogram, which was then used to estimate lead levels at unsampled locations, a process called kriging. Because soil lead levels were spatially correlated, it is likely that a "nonrandom" process generated the lead distribution found. This finding signifies the existence of lead sources which were tentatively identified on historical maps of the area and from past traffic volume patterns. The distribution of kriged estimates of soil lead levels provides an explanatory tool for exploring and identifying potential sources and may be useful for targeting urban soil abatement efforts. PMID- 10677146 TI - Cadmium levels in kidney cortex in Swedish farmers. AB - The cadmium levels in kidney cortex (K-Cd) did not differ statistically between 10 nonsmoking farmers from the south of Sweden, who had a high intake of locally produced food and who were affected by acid precipitation (as indicated by low pH in the drinking water from their private wells) and 10 farmers less affected (medians: K-Cd, 18 vs. 14 microg/g; water pH, 5.2 vs. 7.8). Neither did 10 farmers selected because of "high" blood cadmium (B-Cd) differ from 10 with "low" [medians: K-Cd, 15 vs. 9 microg/g; B-Cd, 2.6 vs. 1.3 nmol/L (0.29 vs. 0.14 microg/L)]. In all 40 farmers, there was an increase of urinary cadmium levels (U Cd) with decreasing drinking water pH (r(s) = -0.32, P = 0.045). Further, K-Cd increased with rising B-Cd (r(s) = 0.33, P = 0.037), and both B-Cd (r(s) = 0.73, P = 0.0005), and U-Cd (r(s) = 0.74, P = 0.0005) rose with increasing age. Further, there was an association between U-Cd and B-Cd (r(s) = 0.68, P = 0.0005). We could not demonstrate with certainty any effect of the acid precipitation on the cadmium retention in the farmers, although the association between U-Cd and drinking water pH deserves further study. PMID- 10677147 TI - Acquisition and retention of lead by young children. AB - The concentrations and isotope ratios of lead in blood and urine, on the hands, and in duplicate diet samples were measured for children living in Omaha, Nebraska. One group consisted of 22 children followed from birth to between 1 and 2 years of age and another group was 20 2- to 4-year-old children followed for 1 year, although some in each group were followed for periods between 3 and 4 years. At no time in life was a component of dietary lead identified in blood by isotope ratios, and blood lead appears dominated by lead derived from the hands, which in turn appears derived from the floors. For some homes floor lead appeared to be a mixture of lead from window sills and from the exterior. Only 2 of the children appear to have ingested lead directly from window sills. Several who lived in homes being remodeled were exposed to lead before the age of 2 years. For those who had been briefly exposed during professional remodeling the blood lead fell with a half-life of 10 months but for those who had suffered prolonged exposure during remodeling by parents the apparent half-life was longer, between 20 and 38 months. PMID- 10677149 TI - Re: evaluating mercury exposure through fish consumption in the Amazon--the roles of biomarkers and predictive models. PMID- 10677148 TI - Maternal bone lead contribution to blood lead during and after pregnancy. AB - We examined bone lead contribution to blood lead in a group of 311 immigrant women, 99% from Latin America, during the third trimester of pregnancy and 1 to 2 months after delivery. We measured in vivo tibia and calcaneus (heel) bone lead concentration in the postdelivery period with K shell X-ray fluorescence. Prenatal and postnatal geometric mean (range) blood lead level was 2.2 microg/dL (0.4 to 38.7) and 2.8 microg/dL (0.4 to 25.4), reflecting low current exposure. Postnatal blood lead level was significantly higher than prenatal (P<0.0001). Mean (range) tibia and calcaneus lead concentration was 6.7 microg/g (-33.7 to 62.2) and 8.4 microg/g (- 30.1 to 66.4), reflecting varying but elevated past lead exposure. Mean calcaneus lead concentration was significantly higher than mean tibia lead concentration (P = 0.055). Variance-weighted multiple regression and structural equation models showed that both calcaneus and tibia lead were directly associated with prenatal blood lead but only calcaneus lead was associated with postnatal blood lead. Increasing natural log years in the United States independently predicted decreasing calcaneus and third-trimester blood lead. The data suggest that while some exogenous lead sources and modulators of blood lead level, such as use of lead-glazed pottery and calcium in the diet, control lead exposure during and after pregnancy, endogenous lead sources from past exposure before immigration continue to influence blood lead levels in this sample. PMID- 10677151 TI - Tissue engineered artificial skin composed of dermis and epidermis. AB - We made an artificial skin comprised of a stratified layer of keratinocytes and a dermal matrix with a type I collagen containing fibroblasts. In this work, we showed keratinocyte behavior under primary culture, gel contractions varying with concentration of collagen solution, and cell growth plots in the collagen gel. The optimum behavior of dermal equivalent could be obtained using 3.0 mg/ml collagen solution and attached gel culture. The attached gel culture had a jumping effect of growth factor on cell growth at the lag phase. To develop the artificial skin, 1x10(5) cells/cm2 of keratinocytes were cultured on the dermal equivalent at air-liquid interface. Finally, to overcome the problem that artificial skin of collagen gel was torn easily during suturing of grafting, we prepared histocompatible collagen mesh and attached the mesh to the bottom of the gel. Cultured artificial skins were successfully grafted onto rats. PMID- 10677150 TI - Influence of different hemodialysis membranes on red blood cell susceptibility to oxidative stress. AB - Oxidative stress is crucial in red blood cell (RBC) damage induced by activated neutrophils in in vitro experiments. The aim of the study was to evaluate whether the bioincompatibility phenomena occurring during hemodialysis (HD) (where neutrophil activation with increased free radical production is well documented) may have detrimental effects on RBC. We evaluated RBC susceptibility to oxidative stress before and after HD in 15 patients using Cuprophan, cellulose triacetate, and polysulfone membrane. RBC were incubated with t-butyl hydroperoxide as an oxidizing agent both in the presence and in the absence of the catalase inhibitor sodium azide. The level of malonaldehyde (MDA), a product of lipid peroxidation, was measured at 0, 5, 10, 15, and 30 min of incubation. When Cuprophan membrane was used, the MDA production was significantly higher after HD, indicating an increased susceptibility to oxidative stress in comparison to pre-HD. The addition of sodium azide enhanced this phenomenon. Both cellulose triacetate and polysulfone membranes did not significantly influence RBC susceptibility to oxidative stress. Neither the level of RBC reduced glutathione nor the RBC glutathione redox ratio changed significantly during HD with any of the membranes used. The RBC susceptibility to oxidative stress was influenced in different ways according to the dialysis membrane used, being increased only when using the more bioincompatible membrane Cuprophan, where neutrophil activation with increased free radical production is well documented. The alterations found in this study might contribute to the reduced RBC longevity of HD patients where a bioincompatible membrane is used. PMID- 10677152 TI - Immobilization of ligand-modified polyamidoamine dendrimer for cultivation of hepatoma cells. AB - Cationic polyamidoamine dendrimers are known to be highly branched cascade polymers. The core part of these polymers, tris(2-aminoethyl)amine, was immobilized onto polystyrene plates to which animal cells do not adhere. using photoreactive 4-(3-trifluoromethylazirino) benzoyl-N-succinimide (TDBA-OSu). Cells of a rat hepatoma cell line, H4-II-E-C3, adhered to a surface immobilized with a first-generation dendrimer probably through interactions between the terminal amino groups of the dendrimer and the cell membranes. The adhered cells were viable, could proliferate, and exhibited urea synthetic activity. The modification of the terminal amino groups with fructose increased the final number of cells obtained after 5 days of cultivation. Multigeneration dendrimers were prepared by repeated linkage of tris(2-aminoethyl)amine with the amino groups. Theoretically, the number of terminal amino groups available for ligand modification is twice as much for each generation of dendrimer growth. Cells cultivated on multigeneration fructose-modified dendrimers exhibited enhanced urea synthetic activity. The use of ligand-modified dendrimers is, therefore, considered to be very promising for the construction of bioartificial organs based on cultivation of the animal cells. PMID- 10677153 TI - Small diameter vascular prosthesis with a nonthrombogenic phospholipid polymer surface: preliminary study of a new concept for functioning in the absence of pseudo- or neointima formation. AB - The purpose of this study was to prepare a small diameter vascular prosthesis functioning without pseudointima formation. A nonthrombogenic phospholipid polymer, the 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, has a cell membrane-like structure and has demonstrated strong nonthrombogenicity. We have recently prepared 2 kinds of vascular prostheses, 2 mm in diameter, composed of the MPC polymer and segmented polyurethane (SPU). One includes 7.5 wt% MPC polymer (SPU/MPC[7.5] prosthesis), and the other includes 10.0 wt% (SPU/MPC[10] prosthesis). These prostheses were placed in rabbit carotid arteries and were retrieved at 1 and 4 weeks after implantation. A pseudointima was observed at 4 weeks on the SPU/MPC(7.5). For the SPU/MPC(10), the surface was macroscopically clear without a pseudointima even after a 4 week implantation. It appears that the SPU/MPC(10) prosthesis, functioning without a pseudointima, possesses a stronger nonthrombogenicity and would be more applicable for clinical use. PMID- 10677154 TI - Modeling flow effects on thrombotic deposition in a membrane oxygenator. AB - The hypothesis that regions of low blood velocity in a membrane oxygenator, as predicted by computational fluid dynamics (CFD), would correspond with regions of clinical thrombotic deposition was investigated. Twenty heparin-coated oxygenators were sectioned following use in adult extracorporeal membrane oxygenation. The activated clotting time (ACT) was maintained at approximately 180 s via heparin infusion throughout the support period. Cross-sections were systematically photographed, and slides made to allow image projection upon a digitizing pad. Thrombotic deposition was traced to allow creation of a device cross-section image with an overlaid color scale representing thrombotic deposition frequency. A two-dimensional CFD model was developed to predict blood velocities throughout the oxygenator cross-section. Direct spatial comparisons were made between maps of CFD modeled blood speed and thrombotic deposition. Theoretical oxygenator design modification was performed within the CFD model to investigate flow paths which might minimize regions of low blood velocity. CFD results demonstrated that low velocity regions qualitatively matched regions with a high incidence of thrombotic deposition. Thrombotic deposition was also correlated to longer perfusion periods. This technique of coupling clinical data and CFD offers the potential to relate flow characteristics to thrombotic deposition and represents a potentially powerful new methodology for the optimization of oxygenator flow-related biocompatibility. PMID- 10677155 TI - Enhanced responsiveness of circulatory neutrophils after cardiopulmonary bypass: increased aggregability and superoxide producing capacity. AB - Cardiac surgery with cardiopulmonary bypass (CPB) induces a whole body inflammatory response that sometimes leads to postoperative organ dysfunction, and neutrophil activation plays an important role in this reaction. Neutrophil priming has been described as a change in neutrophil status such that neutrophils show enhanced responsiveness to a second activating stimulus. We hypothesized that neutrophil priming occurs by cardiac surgery with CPB and is temporally related to the neutrophilia after surgery. To evaluate primed circulatory neutrophil status, we measured aggregation activity stimulated by N-formyl-methyl leucyl-phenyl-alanine (FMLP) and free radical producing activity by tumor necrosing factor (TNF) alpha in peripheral blood samples. Eleven adult patients undergoing elective cardiac surgery with CPB were studied. Blood samples were taken before surgery, at the end of bypass, 12 h after surgery, and 7 days after surgery. Aggregation activity and superoxide generation were significantly elevated 12 h after surgery when compared to presurgery values, indicating that cardiac surgery is associated with circulatory neutrophil priming. The number of neutrophils markedly increased at the end of cardiopulmonary bypass and reached a peak 12 h after surgery. The circulatory neutrophils of cardiac surgical patients become primed after surgery, coincident with the peak neutrophil count. These results suggest that circulatory neutrophils after cardiac surgery with CPB have enhanced responsiveness and are predisposed to systemic inflammation. PMID- 10677156 TI - Pressure drop, shear stress, and activation of leukocytes during cardiopulmonary bypass: a comparison between hollow fiber and flat sheet membrane oxygenators. AB - The membrane oxygenator is known to be superior to the bubble oxygenator, but little information is available about the difference between the hollow fiber and flat sheet membrane oxygenators with regard to pressure drop, shear stress, and leukocyte activation. In this study, we compared these 2 types of membrane oxygenators in patients undergoing cardiopulmonary bypass (CPB) surgery with special focus on leukocyte activation and pressure drop across the oxygenators. Plasma concentration of elastase, a marker indicating leukocyte activation, increased to 593+/-68% in the flat sheet oxygenator group versus 197+/-42% in the hollow fiber oxygenator group (p<0.01) at the end of CPB compared to their respective baseline concentrations before CPB. Pressure drop across the oxygenator was significantly higher in the flat sheet group than in the hollow fiber group throughout the entire period of CPB (p<0.01). High pressure drop across the oxygenator as well as the calculated shear stress was positively correlated with the release of elastase at the end of CPB (r = 0.760, p<0.01, r = 0.692, p<0.01). However, this positive correlation existed in the flat sheet oxygenator but not in the hollow fiber oxygenator. Clinically, both membrane oxygenators have satisfactory performance in O2 and CO2 transfer. These results suggest that a higher pressure drop across the flat sheet oxygenator is associated with more pronounced activation of leukocytes in patients undergoing cardiopulmonary bypass. PMID- 10677157 TI - Maintenance of blood heparin concentration rather than activated clotting time better preserves the coagulation system in hypothermic cardiopulmonary bypass. AB - In cardiopulmonary bypass (CPB), despite heparin regimens in which the activated clotting time (ACT) is kept at more than 400 s, there is biochemical evidence of thrombin generation indicating activation of the coagulation system and increased fibrinolytic activity. Therefore, to reduce the coagulant activation has been one of the main issues in the improvement of CPB. The purpose of this study was to compare the heparin concentration with the ACT and to evaluate the effect of keeping higher heparin concentration on the coagulation and fibrinolytic systems during hypothermic CPB, employing moderate hypothermia (MHT) or deep hypothermic circulatory arrest (DHT). Heparin was either administered to maintain an ACT >400 s (ACT group) or to maintain a whole blood heparin concentration of 3 mg/kg (heparin group). At the lowest core temperature during CPB, the ACT and the heparinase ACT (unrelated to heparin concentration) were increased the most whereas the whole blood heparin concentration was less than half the initial concentration in both ACT groups of MHT and DHT. The thrombin-antithrombin III (TAT) content just after CPB in both MHT and DHT was significantly lower in the heparin group than in the ACT group. In conclusion, ACT does not reflect the whole blood heparin concentration during hypothermic CPB. Furthermore, maintenance of the higher heparin concentration during hypothermic CPB may suppress the activation of the coagulation system via thrombin inhibition. That effect was more remarkable in deep hypothermic CPB. Therefore, we believe that anticoagulation management during hypothermic CPB should be based on the maintenance of the higher blood heparin concentration. PMID- 10677158 TI - Hydrodynamic evaluation of three artificial aortic valve chambers. AB - The effect of chamber geometry on the characteristics of turbulent steady flow through a newly designed artificial heart valve, "the jellyfish valve," has been investigated for flow rates matching those of peak systole. Laser Doppler Anemometry (LDA) was employed to determine the velocity and shear stress distributions at various locations downstream of the jellyfish valve. Three geometrically different aortic valve chambers have been investigated: namely, a chamber with sinuses of Valsalva, an ellipsoidal chamber, and a cylindrical chamber. The results of this investigation indicated that the aorta with sinuses of Valsalva model had the highest turbulent shear stresses whereas the ellipsoidal model gave the highest-pressure drops. However, for the various flow rates examined, including the systole peak value of 26 L/min, it appears that the ellipsoidal model displays better hydrodynamic characteristics in terms of shear stress and uniformity of axial velocity distributions downstream of the jellyfish valve. PMID- 10677159 TI - Evaluation of the inner-surface morphology of an artificial heart by acoustic microscopy. AB - The total artificial heart (TAH) is being developed for permanent replacement of the natural heart instead of heart transplantation. The need for detecting the material fatigue in the TAH is increasing in order to guarantee long-term use. In this study, the inner surface morphology of the TAH was evaluated by a specially developed scanning acoustic microscope (SAM) system operating in the frequency range of 100-200 MHz. The inner sac of our TAH consisted of polyvinylchloride coated with polyurethane, and the SAM investigations were performed before and after the implantations in goats. The amplitude images of the SAM demonstrated protein adhesion on the inner surface of the TAH after the animal experiment, and the phase images showed distortion of the wall with spatial resolution of 0.2 microm. These results suggest the feasibility of a high-frequency ultrasound for evaluating the material fatigue of TAH. PMID- 10677160 TI - Significance of percutaneous cardiopulmonary bypass support for volume reduction surgery with severe hypercapnia. AB - In patients with reduced respiratory function, lung resection is associated with high risk because separate ventilation is generally needed for safe management. For patients with end-stage emphysema, intraoperative respiratory management is important and particularly difficult because neither incomplete oxygenation nor selective ventilation can be performed, so the operation may be interrupted. In this study, we assess the effectiveness of the percutaneous cardiopulmonary support (PCPS) system for lung volume reduction surgery in patients with severe hypercapnia (arterial carbon dioxide tension >50 mm Hg) and discuss the significance of PCPS for patients who are beyond the standard criteria for lung volume reduction surgery (LVRS). We studied 3 patients with severe hypercapnia due to emphysema who underwent volume reduction surgery. One patient was previously treated surgically for contralateral pneumothorax. All patients had a severe smoking history and were suspected to have fragile lungs. During the operation. PCPS provided sufficient support flow. Intraoperative management using PCPS was easy, and no severe complications were observed. One patient exhibited severe hemodynamic deterioration on postoperative Day 15. Other patients' PaCO2 improved postoperatively. One had a calcification of a femoral artery, but there was no trouble inserting a cannula. Bilateral or unilateral volume reduction surgery was performed under PCPS in patients with end-stage emphysema. We conclude that PCPS is an adjunct to LVRS, useful for intraoperative management of some patients with severe hypercapnea, and the LVRS indications can be extended. PMID- 10677161 TI - Five-year follow-up of valve replacement with the Jyros bileaflet mechanical valve. AB - Jyros bileaflet rotating valves were implanted as a clinical trial conducted in Japan, and the 5-year results were assessed. Nineteen patients underwent implantation of the valves: 14 in the mitral and 5 in the aortic position. The mean follow-up period was 65.4+/-15.7 months. There was 1 case of late death due to fatal arrhythmia and another case of cerebral thromboembolism (1.0% per patient year). All survivors were in New York Heart Association class I. On the early postoperative cinefluorography, 8 valves (42.1%) showed rotation of the leaflets. However, in the latest assessment 6 valves (33.3%) showed rotation, some valves had stopped rotation, and others had started to rotate during the follow-up period. The Jyros valve functions effectively, similar to other bileaflet valves. However, because the correlation between thromboembolism and the rotation mechanism is not clear, further follow-up of our patients and more implant studies are necessary to elucidate this issue. PMID- 10677162 TI - A clinical use of the Kyocera Gyro C1E3 centrifugal pump. AB - The Kyocera Gyro C1E3 centrifugal blood pump was clinically applied for a cardiopulmonary bypass (CPB) of coronary artery bypass grafting (CABG). The patient was 72-year-old male with postinfarction unstable angina. The surgery was carried out on November 20, 1998. The air inside the pump was easily and quickly removed, and its controllability was excellent. The pump flow during operation was maintained 2.2 L/m2. Total CPB time was 173 min. Perioperative parameters of hemolysis and cytotoxicity were not remarkably changed. Macroscopically and microscopically, there were no thrombi inside the pump after usage. This is the first reported case of clinical use of the Kyocera Gyro C1E3 pump. PMID- 10677163 TI - Epidemiology and molecular genetics of colorectal cancer. AB - Colorectal cancer is among the most common cancers affecting the western world. By the age of 70 yr, at least 50% of the Western population will develop some form of colorectal tumor, spanning the spectrum from an early benign polyp to an invasive adenocarcinoma. It is estimated that approximately 10% of the benign polypoid lesions will progress to invasive carcinoma. The concept that serial genetic changes are responsible for the transition from benign to neoplastic disease is not new. The description of hereditary cancers and the demonstration of carcinogenic substances inducing DNA damage have provided the foundation for the field of molecular oncology. During the past three decades, our understanding of how genetic alterations culminate in cancer has progressed rapidly, though the complete process has not been fully defined. The research to date has spanned many oncologic diseases, but has been especially well defined in colorectal cancer. The knowledge of the genetic alterations that result in colorectal cancer has important ramifications for future prevention, detection, and treatment of this disease. PMID- 10677164 TI - Colorectal cancer screening and follow-up. AB - Cancer of the colon and rectum is a significant health problem in the United States. Nearly 50% of the 186,000 patients diagnosed annually with colorectal cancer will eventually die of their disease. Because development of a colorectal carcinoma is most frequently preceded by the development of a well-recognized pre malignant lesion, screening modalities can significantly impact the incidence and mortality rate of this disease. Population screening employing digital rectal examination, fecal occult blood testing and endoscopic examination of the rectum and colon has been demonstrated to reduce the risk of death from colorectal cancer. Screening regimens should be instituted at an earlier age and with increased frequency for patients in the highest risk categories. Patients who have been treated for a cancer of the colon or rectum should undergo surveillance at regular intervals in an attempt to identify recurrences of disease both in the residual colon and rectum and at distant sites. Most physicians and patients believe that intensive follow-up strategies will afford improved survival and quality of life, however few randomized studies examining the utility of intensive follow-up programs have been performed and the quality of cancer related follow-up literature is generally poor. Good-quality clinical trials are needed to sort out which tests make a difference in the patient's long-term outcome. The algorithm for surveillance for recurrence in the future may be altered as newer testing modalities are developed. PMID- 10677165 TI - Tumor markers--prognostic and therapeutic implications for colorectal carcinoma. AB - Molecular prognostic markers are molecules produced by either the tumor or the host (patient) whose expression is associated with the clinical outcome. Three types of molecular markers exist that characterize different aspects of the tumor : host relationship: (1) tumor biology, (2) tumor burden and (3) host response. The first type of marker is measured within the primary or metastatic tumor mass and defines the aggressiveness of the cancer and its ability to respond to therapy. The other two types of markers are usually measured in the blood and assess concentrations of circulating tumor products or cytokines that may be involved in host resistance to the cancer. In this brief review we will define each type of marker, provide examples of their current utility and then describe how these markers may be useful. PMID- 10677166 TI - Surgical management of primary colorectal cancer. AB - Surgery for potentially curable colorectal cancer most commonly involves resection of the primary tumor and regional lymph nodes. However, the site, extent and presentation of disease have an impact on surgical strategy and the use of combined modality therapy. For colon cancer, complex presentations such as obstructing or perforated colon cancer may influence surgical therapy, and issues pertaining to en bloc resection and oophorectomy remain unresolved. For rectal cancer, surgical management may range from local excision to radical resection. Extent of resection and relatively new operative techniques such as coloanal anastomosis with or without a colonic pouch reservoir are directed towards optimizing both oncologic and functional results. PMID- 10677167 TI - Minimally invasive techniques for colorectal cancer. AB - Laparoscopic approaches are increasingly being applied to colorectal surgical procedures. Initial concerns regarding the existence of benefits from the laparoscopic approach have now been addressed. Even as these were being addressed, however, further concerns arose regarding the appropriateness of this technique in malignancy. Colorectal cancer is the only intra-abdominal malignancy that is knowingly resected employing laparoscopic techniques. This controversy was highlighted by reports of early wound implants. With careful technique, training and experience, however, wound recurrences are rarely seen, suggesting that this phenomenon, in the clinical setting, is primarily technique-related. Lack of rigorous evidence either condemning or supporting the laparoscopic approach for colorectal cancer resulted in the establishment of several large scale randomized, prospective trials, all currently in progress, that aim to determine if laparoscopic resection of colorectal cancer results in oncologic outcomes comparable to the open approach. PMID- 10677168 TI - Adjuvant and neoadjuvant chemoradiation therapy for primary colorectal cancer. AB - The management of rectal cancer presents substantial challenges. Patients with T3 and/or node-positive rectal cancers are at high risk for local failure and distant metastases (DM). Adjuvant radiation has been shown to decrease local recurrence (LR) rates; however, this local therapy has not been demonstrated to improve survival when compared to surgery alone. In several prospective randomized trials adjuvant chemoradiation with 5-fluorouracil-(5-FU)-based chemotherapy improved LR rates, DM rates, and overall survival (OS). The optimal chemotherapeutic regimen has not been determined; however, studies comparing standard IV bolus 5-FU administration with continuous infusion (CI) 5-FU demonstrated that CI administration was superior. Preoperative therapy has potential advantages over adjuvant therapy such as less acute bowel toxicity and improved sphincter preservation. Preoperative chemoradiation has been shown in several studies to improve LR rates and OS when compared to surgery alone. Our current approach to patients with resectable T3 or N1 cancer in the distal two thirds of the rectum on preoperative staging is preoperative chemoradiation with planned postoperative chemotherapy. This regimen offers the best chance for local control and disease-free survival while potentially downstaging the tumor and improving sphincter preservation. PMID- 10677169 TI - Molecular determinants of colon cancer metastasis. AB - Colon cancer metastasis is a tightly regulated process that requires a cancer cell to express genes that allow progression through various distinct steps. Aberrations in gene expression by cancer cells leads to transformation, growth, angiogenesis, invasion, dissemination and survival in the circulation, attachment in the organ of metastasis, and again invasion, growth, and angiogenesis. In addition to the genotype/phenotype of the tumor cell, for a tumor cell to become a clinically relevant metastasis, it must be able to respond appropriately to the environment. This includes being able to utilize growth factors and blood vessels from the organ of metastasis for the benefit of the tumor mass. Understanding the molecular and biologic mechanisms of colon cancer metastasis will allow the development of rationale therapeutic strategies that are more likely to impact the natural history of this disease than current therapies. PMID- 10677170 TI - Multidisciplinary management of metastatic colorectal cancer. AB - When colorectal cancer metastasizes to distant organs, usually multiple sites are involved and treatment consists primarily of systemic chemotherapy and supportive care. Chemotherapeutic agents effective against metastatic colorectal cancer include 5-fluorouracil, often used in combination with leucovorin or methotrexate, and irinotecan (CPT-11). Median survival with optimal chemotherapy regimens ranges from 10 to 15 months. Less frequently, colorectal cancer metastasizes only to the liver or lung. In a minority of these cases, surgical resection can be performed and results in a median survival of 28-46 months for hepatic resections and 24-25 months for pulmonary resections. Five-year survival rates range from 24 to 38% and 21 to 44% for hepatic and pulmonary resections, respectively. For isolated liver metastases that are not surgically resectable, other regional therapies that can be considered are hepatic cryosurgery, radiofrequency ablation, and hepatic arterial infusion chemotherapy. Median survival following cryosurgery is between 26 and 30 months, while median survival following radiofrequency ablation has not been established in large series. Hepatic arterial infusion chemotherapy, especially with newer combination drug regimens, may increase survival in patients with isolated liver metastases compared to systemic chemotherapy, but this must be confirmed in randomized, prospective trials. Colorectal cancer metastases to the brain can be treated with radiation therapy or surgical resection, but median survival with treatment is less than one year. PMID- 10677171 TI - Multidisciplinary management of recurrent colorectal cancer. AB - Isolated pelvic recurrence of rectal carcinoma may occur in up to one third of patients following definitive resection of primary disease. The means by which recurrence is diagnosed, methods by which it may be treated, and reported outcomes are all evolving and improving. Current data indicate that a substantial proportion of patients treated by aggressive multi-modality salvage therapy may be provided with durable survival. This review highlights current concepts in the diagnosis and management of locally recurrent rectal carcinoma. PMID- 10677172 TI - [Etiopathogeny of fetal macrosomia]. AB - Fetal macrosomia is an heterogeneous condition in terms of definition and etiologic factors. Recent findings suggest that macrosomia should not be classified on the basis of birth weight and gestational age alone. The ponderal index delineates a symmetric and an asymmetric subtype of macrosomia. The relationship between maternal diabetes and fetal macrosomia has been extensively investigated. However, eighty percent of macrosomic infants are born to mothers who are not hyperglycemic, and various factors have been associated. Maternal factors explain approximately 50% of the variance in birth weight, whereas paternal factors have no significant effect. The predisposition to excessive fetal growth may be shared within the intra uterine environment and the fetal genome. The respective roles of lipids, amino acids, hormones such as leptin, and growth factors need to be evaluated. Perinatal morbidity and long term consequences such as obesity and glucose intolerance might be associated with some of the factors leading to fetal overgrowth. PMID- 10677173 TI - [Psychotropes and pregnancy]. PMID- 10677180 TI - Dental fillings. The silver in them thar molars. PMID- 10677181 TI - Mental health. Adult attention deficit hyperactivity disorder. PMID- 10677182 TI - Health beat. Targeted chemotherapy. PMID- 10677184 TI - Caregiving. The health hazards of helping. PMID- 10677183 TI - By the way, doctor. I lost my sense of taste and smell about four years ago. I like to eat - always have - and I do what I can to keep on enjoying it by paying attention to how the food looks, how it feels in my mouth, and my memories of it. But it isn't the same. Is there anything that can be done? I'm 83 and in good health. PMID- 10677185 TI - General review: personality disorders - part I. PMID- 10677186 TI - Insights: emotional illness and creativity: a psychoanalytic perspective. PMID- 10677187 TI - Is ritalin underused? PMID- 10677188 TI - Gay youth at risk. PMID- 10677189 TI - Forum. How should suicidal behavior be managed? PMID- 10677190 TI - The mortality of men in Russia: a cautionary tale. PMID- 10677191 TI - To err is human, but... PMID- 10677192 TI - Sexuality. It takes two: coping with erectile dysfunction. PMID- 10677194 TI - Lyme disease update. PMID- 10677193 TI - Feet. Taking steps toward good health. PMID- 10677195 TI - By the way, doctor. I recently read that a test for the virus linked to cervical cancer may be better than a Pap smear. Does this mean I can avoid a regular pelvic exam and Pap smear? Should I ask my doctor for this test? PMID- 10677200 TI - Health beat. Vitamin E: E for exaggerated? PMID- 10677201 TI - By the way, doctor. I went through menopause about 10 years ago and have been taking estrogen therapy ever since. My uterus was removed because of fibroids, so there is no risk of endometrial cancer, which is why I am taking estrogen without progesterone. Should I have a bone density scan? A lot of my friends are having them, but I'm not sure how this test could alter my therapy. PMID- 10677202 TI - Nalmefene for alcohol dependence. PMID- 10677203 TI - GERD: more than just heartburn. PMID- 10677204 TI - The healthiness of the long-distance runner. PMID- 10677205 TI - On call: I am a 68-year-old man who had bypass surgery last month. My heart did fine, but I had trouble urinating after my urinary catheter came out. The problem cleared up in a day, but my doctor ordered a PSA test, which came back high (7.8). I don't feel up to another operation. What should I do? PMID- 10677206 TI - The spasmodic peptide defines a new conotoxin superfamily. AB - We purified and characterized a peptide from the venom of Conus textile that makes normal mice assume the phenotype of a well-known mutant, the spasmodic mouse. This "spasmodic" peptide has 27 amino acids, including two gamma carboxyglutamate (Gla) residues. A cDNA clone encoding the precursor for the peptide was identified; a gamma-carboxylation recognition signal sequence (gamma CRS) is present in the -1 --> -20 region of the peptide precursor. Both the gamma CRS and the position of the Gla residues in the mature toxin are notably different from other Gla-containing conopeptides. The spasmodic peptide has a novel disulfide framework and distinct signal sequence which together define a new P-superfamily of conopeptides. A cDNA encoding another member of the P superfamily was identified from a different species, Conus gloriamaris. PMID- 10677207 TI - Crystal structure of the second domain of the human copper chaperone for superoxide dismutase. AB - The human copper chaperone for superoxide dismutase (hCCS) delivers the essential copper ion cofactor to copper,zinc superoxide dismutase (SOD1), a key enzyme in antioxidant defense. Mutations in SOD1 are linked to familial amyotrophic lateral sclerosis (FALS), a fatal neurodegenerative disorder. The molecular mechanisms by which SOD1 is recognized and activated by hCCS are not understood. To better understand this biochemical pathway, we have determined the X-ray structure of the largest domain of hCCS (hCCS Domain II) to 2. 75 A resolution. The overall structure is closely related to that of its target enzyme SOD1, consisting of an eight-stranded beta-barrel and a zinc-binding site formed by two extended loops. The first of these loops provides the ligands to a bound zinc ion, and is analogous to the zinc subloop in SOD1. The second structurally resembles the SOD1 electrostatic channel loop, but lacks many of the residues important for catalysis. Like SOD1 and yCCS, hCCS forms a dimer using a highly conserved interface. In contrast to SOD1, however, the hCCS structure does not contain a copper ion bound in the catalytic site. Notably, the structure reveals a single loop proximal to the dimer interface which is unique to the CCS chaperones. PMID- 10677208 TI - Hybrid and complex glycans are linked to the conserved N-glycosylation site of the third eight-cysteine domain of LTBP-1 in insect cells. AB - Covalent association of LTBP-1 (latent TGF-beta binding protein-1) to latent TGF beta is mediated by the third eight-cysteine (also referred to as TB) module of LTBP-1, a domain designated as CR3. Spodoptera frugiperda (Sf9) cells have proved a suitable cell system in which to study this association and to produce recombinant CR3, and we show here that another lepidopteran cell line, Trichoplusia niTN-5B1-4 (High-Five) cells, allows the recovery of large amounts of functional recombinant CR3. CR3 contains an N-glycosylation site, which is conserved in all forms of LTBP known to date. When we examined the status of this N-glycosylation using MALDI-TOF mass spectrometry and enzymatic analysis, we found that CR3 is one of the rare recombinant peptides modified with complex glycans in insect cells. Sf9 cells mainly processed the fucosylated paucomannosidic structure (GlcNAc)(2)(Mannose)(3)Fucose, although hybrid and complex N-glycosylations were also detected. In High-Five cells, the peptide was found to be modified with a wide variety of hybrid and complex sugars in addition to paucomanosidic oligosaccharides. Most glycans had one or two fucose residues bound through alpha1,3 and alpha1,6 linkages to the innermost GlcNAc. On the basis of these results and on the structure of an eight-cysteine domain from fibrillin-1, we present a model of glycosylated CR3 and discuss the role of glycosylation in eight-cysteine domain protein-protein interactions. PMID- 10677209 TI - The NMR structure of the nucleocapsid protein from the mouse mammary tumor virus reveals unusual folding of the C-terminal zinc knuckle. AB - The nucleocapsid protein (NC) from the mouse mammary tumor virus (MMTV) has been overexpressed in Escherichia coli and purified to homogeneity for structural studies by nuclear magnetic resonance (NMR) spectroscopy. The protein contains two copies of a conserved zinc-coordinating "CCHC array" or "zinc knuckle" motif common to the nucleocapsid proteins of nearly all known retroviruses. The residues comprising and adjacent to the zinc knuckles were assigned by standard two-dimensional (1)H and three-dimensional (1)H-(15)N NMR methods; the rotational dynamic properties of the protein were determined from (15)N relaxation experiments, and distance restraints derived from the nuclear Overhauser effect (NOE) data were used to calculate the three-dimensional structure. The (1)H-(1)H NOE and (15)N relaxation data indicate that the two zinc knuckles do not interact with each other, but instead behave as independently folded domains connected by a flexible 13-residue linker segment. The proximal zinc knuckle folds in a manner that is essentially identical to that observed previously for the two zinc knuckles of the human immunodeficiency virus type 1 nucleocapsid protein and for the moloney murine leukemia virus nucleocapsid zinc knuckle domain. However, the distal zinc knuckle of MMTV NC exhibits a rare three-dimensional fold that includes an additional C-terminal beta-hairpin. A similar C-terminal reverse turn like structure was observed recently in the distal zinc knuckle of the Mason Pfizer monkey virus nucleocapsid protein [Gao, Y., et al. (1998) Protein Sci. 7, 2265-2280]. However, despite a high degree of sequence homology, the conformation and orientation of the beta-hairpin in MMTV NC is significantly different from that of the reverse turn in MPMV NC. The results support the conclusion that structural features of NC zinc knuckle domains can vary significantly among the different genera of retroviridae, and are discussed in terms of the recent and surprising discovery that MMTV NC can facilitate packaging of the HIV-1 genome in chimeric MMTV mutants. PMID- 10677210 TI - Crystal structure of adenosine 5'-phosphosulfate kinase from Penicillium chrysogenum. AB - Adenosine 5'-phosphosulfate (APS) kinase catalyzes the second reaction in the two step conversion of inorganic sulfate to 3'-phosphoadenosine 5'-phosphosulfate (PAPS). This report presents the 2.0 A resolution crystal structure of ligand free APS kinase from the filamentous fungus, Penicillium chrysogenum. The enzyme crystallized as a homodimer with each subunit folded into a classic kinase motif consisting of a twisted, parallel beta-sheet sandwiched between two alpha-helical bundles. The Walker A motif, (32)GLSASGKS(39), formed the predicted P-loop structure. Superposition of the APS kinase active site region onto several other P-loop-containing proteins revealed that the conserved aspartate residue that usually interacts with the Mg(2+) coordination sphere of MgATP is absent in APS kinase. However, upon MgATP binding, a different aspartate, Asp 61, could shift and bind to the Mg(2+). The sequence (156)KAREGVIKEFT(166), which has been suggested to be a (P)APS motif, is located in a highly protease-susceptible loop that is disordered in both subunits of the free enzyme. MgATP or MgADP protects against proteolysis; APS alone has no effect but augments the protection provided by MgADP. The results suggest that the loop lacks a fixed structure until MgATP or MgADP is bound. The subsequent conformational change together with the potential change promoted by the interaction of MgATP with Asp 61 may define the APS binding site. This model is consistent with the obligatory ordered substrate binding sequence (MgATP or MgADP before APS) as established from steady state kinetics and equilibrium binding studies. PMID- 10677211 TI - Computational methodology for estimating changes in free energies of biomolecular association upon mutation. The importance of bound water in dimer-tetramer assembly for beta 37 mutant hemoglobins. AB - The computational modeling program HINT (Hydropathic INTeractions), an empirical hydropathic force field that includes hydrogen bonding, Coulombic, and hydrophobic terms, was used to model the free energy of dimer-tetramer association in a series of deoxy hemoglobin beta 37 double mutants. Five of the analyzed mutants (beta 37W --> Y, beta 37W --> A, beta 37W --> G, beta 37W --> E, and beta 37W --> R) have been solved crystallographically and characterized thermodynamically and subsequently made a good test set for the calibration of our method as a tool for free energy prediction. Initial free energy estimates for these mutants were conducted without the inclusion of crystallographically conserved water molecules and systematically underestimated the experimentally calculated loss in free energy observed for each mutant dimer-tetramer association. However, the inclusion of crystallographic waters, interacting at the dimer-dimer interface of each mutant, resulted in HINT free energy estimates that were more accurate with respect to experimental data. To evaluate the ability of our method to predict free energies for de novo protein models, the same beta 37 mutants were computationally generated from native deoxy hemoglobin and similarly analyzed. Our theoretical models were sufficiently robust to accurately predict free energy changes in a localized region around the mutated residue. However, our method did not possess the capacity to generate the long range secondary structural effects observed in crystallographically solved mutant structures. Final method analysis involved the computational generation of structurally and/or thermodynamically uncharacterized beta 37 deoxy hemoglobin mutants. HINT analysis of these structures revealed that free energy predictions for dimer-tetramer association in these models agreed well with previously observed energy predictions for structurally and thermodynamically characterized beta 37 deoxy hemoglobin mutants. PMID- 10677212 TI - Helical interactions in the HIV-1 gp41 core reveal structural basis for the inhibitory activity of gp41 peptides. AB - The HIV-1 gp41 envelope protein mediates membrane fusion that leads to virus entry into the cell. The core structure of fusion-active gp41 is a six-helix bundle in which an N-terminal three-stranded coiled coil is surrounded by a sheath of antiparallel C-terminal helices. A conserved glutamine (Gln 652) buried in this helical interface replaced by leucine increases HIV-1 infectivity. To define the basis for this enhanced membrane fusion activity, we investigate the role of the Gln 652 to Leu substitution on the conformation, stability, and biological activity of the N34(L6)C28 model of the gp41 ectodomain core. The 2.0 A resolution crystal structure of the mutant molecule shows that the Leu 652 side chains make prominent contacts with hydrophobic grooves on the surface of the central coiled coil. The Gln 652 to Leu mutation leads to a marginal stabilization of the six-helix bundle by -0.8 kcal/mol, evaluated from thermal unfolding experiments. Strikingly, the mutant N34(L6)C28 peptide is a potent inhibitor of HIV-1 infection, with 10-fold greater activity than the wild-type molecule. This inhibitory potency can be traced to the corresponding C-terminal mutant peptide that likely has greater potential to interact with the coiled-coil trimer. These results provide strong evidence that conserved interhelical packing interactions in the gp41 core are important determinants of HIV-1 entry and its inhibition. These interactions also offer a test-bed for the development of more potent analogues of gp41 peptide inhibitors. PMID- 10677213 TI - Characterization of a novel DNA primase from the Salmonella typhimurium bacteriophage SP6. AB - The gene for the DNA primase encoded by Salmonella typhimurium bacteriophage SP6 has been cloned and expressed in Escherichia coli and its 74-kDa protein product purified to homogeneity. The SP6 primase is a DNA-dependent RNA polymerase that synthesizes short oligoribonucleotides containing each of the four canonical ribonucleotides. GTP and CTP are both required for the initiation of oligoribonucleotide synthesis. In reactions containing only GTP and CTP, SP6 primase incorporates GTP at the 5'-end of oligoribonucleotides and CMP at the second position. On synthetic DNA templates, pppGpC dinucleotides are synthesized most rapidly in the presence of the sequence 5'-GCA-3'. This trinucleotide sequence, containing a cryptic dA at the 3'-end, differs from other known bacterial and phage primase recognition sites. SP6 primase shares some properties with the well-characterized E. colibacteriophage T7 primase. The T7 DNA polymerase can use oligoribonucleotides synthesized by SP6 primase as primers for DNA synthesis. However, oligoribonucleotide synthesis by SP6 primase is not stimulated by either the E. coli- or the T7-encoded ssDNA binding protein. An amino acid sequence alignment of the SP6 and T7 primases, which share only 22.4% amino acid identity, indicates amino acids likely critical for oligoribonucleotide synthesis as well as a putative Cys(3)His zinc finger motif that may be involved in DNA binding. PMID- 10677214 TI - Mutational, kinetic, and NMR studies of the roles of conserved glutamate residues and of lysine-39 in the mechanism of the MutT pyrophosphohydrolase. AB - The MutT enzyme catalyzes the hydrolysis of nucleoside triphosphates (NTP) to NMP and PP(i) by nucleophilic substitution at the rarely attacked beta-phosphorus. The solution structure of the quaternary E-M(2+)-AMPCPP-M(2+) complex indicated that conserved residues Glu-53, -56, -57, and -98 are at the active site near the bound divalent cation possibly serving as metal ligands, Lys-39 is positioned to promote departure of the NMP leaving group, and Glu-44 precedes helix I (residues 47-59) possibly stabilizing this helix which contributes four catalytic residues to the active site [Lin, J. , Abeygunawardana, C., Frick, D. N., Bessman, M. J., and Mildvan, A. S. (1997) Biochemistry 36, 1199-1211]. To test these proposed roles, the effects of mutations of each of these residues on the kinetic parameters and on the Mn(2+), Mg(2+), and substrate binding properties were examined. The largest decreases in k(cat) for the Mg(2+)-activated enzyme of 10(4.7)- and 10(2.6)-fold were observed for the E53Q and E53D mutants, respectively, while 97-, 48-, 25-, and 14-fold decreases were observed for the E44D, E56D, E56Q, and E44Q mutations, respectively. Smaller effects on k(cat) were observed for mutations of Glu-98 and Lys-39. For wild type MutT and its E53D and E44D mutants, plots of log(k(cat)) versus pH exhibited a limiting slope of 1 on the ascending limb and then a hump, i.e., a sharply defined maximum near pH 8 followed by a plateau, yielding apparent pK(a) values of 7.6 +/- 0.3 and 8.4 +/- 0.4 for an essential base and a nonessential acid catalyst, respectively, in the active quaternary MutT-Mg(2+)-dGTP-Mg(2+) complex. The pK(a) of 7.6 is assigned to Glu-53, functioning as a base catalyst in the active quaternary complex, on the basis of the disappearance of the ascending limb of the pH-rate profile of the E53Q mutant, and its restoration in the E53D mutant with a 10(1.9)-fold increase in (k(cat))(max). The pK(a) of 8.4 is assigned to Lys-39 on the basis of the disappearance of the descending limb of the pH-rate profile of the K39Q mutant, and the observation that removal of the positive charge of Lys-39, by either deprotonation or mutation, results in the same 8.7-fold decrease in k(cat). Values of k(cat) of both wild type MutT and the E53Q mutant were independent of solvent viscosity, indicating that a chemical step is likely to be rate-limiting with both. A liganding role for Glu-53 and Glu-56, but not Glu-98, in the binary E-M(2+) complex is indicated by the observation that the E53Q, E53D, E56Q, and E56D mutants bound Mn(2+) at the active site 36-, 27-, 4.7-, and 1.9-fold weaker, and exhibited 2.10-, 1.50-, 1.12-, and 1.24-fold lower enhanced paramagnetic effects of Mn(2+), respectively, than the wild type enzyme as detected by 1/T(1) values of water protons, consistent with the loss of a metal ligand. However, the K(m) values of Mg(2+) and Mn(2+) indicate that Glu-56, and to a lesser degree Glu-98, contribute to metal binding in the active quaternary complex. Mutations of the more distant but conserved residue Glu-44 had little effect on metal binding or enhancement factors in the binary E-M(2+) complexes. Two-dimensional (1)H-(15)N HSQC and three-dimensional (1)H-(15)N NOESY-HSQC spectra of the kinetically damaged E53Q and E56Q mutants showed largely intact proteins with structural changes near the mutated residues. Structural changes in the kinetically more damaged E44D mutant detected in (1)H-(15)N HSQC spectra were largely limited to the loop I-helix I motif, suggesting that Glu-44 stabilizes the active site region. (1)H-(15)N HSQC titrations of the E53Q, E56Q, and E44D mutants with dGTP showed changes in chemical shifts of residues lining the active site cleft, and revealed tighter nucleotide binding by these mutants, indicating an intact substrate binding site. (ABSTRACT TRUNCATED) PMID- 10677215 TI - Active intracellular domain of Notch enhances transcriptional activation of CCAAT/enhancer binding protein beta on a rat pregnancy-specific glycoprotein gene. AB - Pregnancy-specific glycoproteins (PSGs) are primarily expressed in the placenta and become the major glycoproteins at term. To understand the regulation of PSG expression, we characterized the promoter elements of a rodent PSG gene, rnCGM3, and delineated three nuclear protein binding sites: FPI, -II, and -III in the 5' flanking region of the gene. The FPII-binding factor is shown to be C/EBPbeta, which positively regulates rnCGM3 expression [Chen, H., et al. (1995) DNA Cell Biol. 14, 681-688]. In the current study, we used the yeast one-hybrid system to isolate transcription factors binding to the FPIII site and demonstrated that a rodent J kappa recombination signal sequence binding protein, rRBPJ kappa-2N, bound to the FPIII site. Electrophoretic mobility shift assay with rat placental nuclear proteins revealed a constitutive occupancy of the FPIII site by RBPJ kappa. By transient expression analyses, we demonstrated that rRBPJ kappa-2N repressed the expression from an FPIII-driven SV40 promoter. However, this repression effect was counteracted by the active intracellular domain of Notch (NotchIC). Using the native rnCGM3 promoter construct, we demonstrated that the promoter activity stimulated by C/EBP beta was also repressed by rRBPJ kappa-2N but enhanced by NotchIC. Additionally, we found that NotchIC can stimulate expression through another RBPJkappa site within the FPI site. A functional interaction between factors binding to the FPI, FPII, and FPIII sites is proposed. PMID- 10677216 TI - RPD3-type histone deacetylases in maize embryos. AB - Posttranslational core histone acetylation is established and maintained by histone acetyltransferases and deacetylases. Both have been identified as important transcriptional regulators in various eukaryotic systems. In contrast to nonplant systems where only RPD3-related histone deacetylases (HD) have been characterized so far, maize embryos contain three unrelated families of deacetylases (HD1A, HD1B, and HD2). Purification, cDNA cloning, and immunological studies identified the two maize histone deacetylase HD1B forms as close homologues of the RPD3-type deacetylase HDAC1. Unlike the other maize deacetylases, HD1A and nucleolar HD2, HD1B copurified as a complex with a protein related to the retinoblastoma-associated protein, Rbap46. Two HD1B mRNA species could be detected on RNA blots, encoding proteins of 58 kDa (HD1B-I) and 51 kDa (HD1B-II). HD1B-I (zmRpd3) represents the major enzyme form as judged from RNA and immunoblots. Levels of expression of HD1B-I and -II mRNA differ during early embryo germination; HD1B-I mRNA and protein are present during the entire germination pathway, even in the quiescent embryo, whereas HD1B-II expression starts when meristematic cells enter S-phase of the cell cycle. In line with previous results, HD1B exists as soluble and chromatin-bound enzyme forms. In vivo treatment of meristematic tissue with the deacetylase inhibitor HC toxin does not affect the expression of the three maize histone deacetylases, whereas it causes downregulation of histone acetyltransferase B. PMID- 10677217 TI - Optical spectroscopic study of the effects of a single deoxyribose substitution in a ribose backbone: implications in RNA-RNA interaction. AB - The 2'-OH group in the ribose sugars of an RNA molecule plays an important role in guiding tertiary interactions that stabilize different RNA structural motifs. Deoxyribose, or 2'-OH by 2'-H, substitution in both the single-stranded and the duplex part of an RNA backbone has been routinely used to evaluate what role the 2'-OH plays in different tertiary interactions that guide an RNA-RNA contact. A deoxyribose substitution not only has the effect of removing a hydrogen bond donating group, but also introduces a sugar moiety with a preference for C2'-endo pucker in a backbone of predominantly C3'-endo sugars. This study evaluates the effects of a single deoxyribose substitution in both single-stranded and double helical forms of RNA oligomers. A single-stranded, nonrepetitive 7-mer oligoribonucleotide (7-mer RNA) and four different variants having the same base sequence but with a single deoxyribose sugar at different positions in the strands have been studied by ultraviolet (UV) absorption, circular dichroism (CD), and Fourier transform infrared (FTIR) spectroscopy. Duplexes were formed by association with the complementary strand of the 7-mer RNA. The results show that both RNA and DNA single strands have preorganized conformations with spectral properties resembling those of A- and B-form helices, respectively, with RNA being more heterogeneous than its DNA counterpart. A single deoxyribose substitution perturbs the structure of the RNA backbone, with the effect being more pronounced in the single-stranded than in the duplex structure. The perturbation depends on the position of the 2'-H substitution in the strand. PMID- 10677218 TI - Human mitochondrial DNA polymerase holoenzyme: reconstitution and characterization. AB - We have reconstituted the holoenzyme of the human mitochondrial DNA polymerase from cloned and overexpressed catalytic and accessory subunits. We have examined the polymerization activity of the catalytic subunit alone and of the holoenzyme to establish the function of the accessory subunit in this two subunit enzyme. The accessory subunit associates with the catalytic subunit with a dissociation constant of 35 +/- 16 nM as measured by the concentration dependence of its effect in stimulating maximal DNA binding and polymerization. At saturating concentrations, the accessory subunit contributes to every kinetic parameter examined to facilitate tighter binding of DNA and nucleotide and faster replication. The accessory protein makes the DNA binding 3.5-fold tighter (K(d) of 9.9 +/- 2.1 nM compared to 39 +/- 10 nM for the catalytic subunit alone) without significantly affecting the DNA dissociation rate (0.02 +/- 0.001 compared to 0.03 +/- 0.001 s(-)(1)). The ground-state nucleotide binding is improved from 4.7 +/- 2.0 to 0.78 +/- 0.065 microM, and the maximum DNA polymerization rate is increased from 8.7 +/- 1.1 to 45 +/- 1 s(-)(1) by the addition of the accessory protein. This leads to an increase in processivity from an estimated 290 +/- 46 to 2250 +/- 162. Although the accessory protein has been described as a "processivity factor" because of its effect on the ratio of rate constants defining processivity, this terminology falls short of adequately describing the profound effects of the small subunit on nucleotide-binding and incorporation catalyzed by the large subunit. By using the complete holoenzyme, we can now proceed with a comprehensive analysis of the structural and mechanistic determinants of enzyme specificity that govern toxicity of nucleoside analogues used in the treatment of viral infections such as AIDS. PMID- 10677219 TI - Activation of NF-kappa B and p38 MAP kinase is not sufficient for triggering efficient HIV gene expression in response to stress. AB - Recent studies have established an essential role for p38 MAP kinase in UV activation of human immunodeficiency virus (HIV) gene expression. However, p38 MAP kinase is not involved in activation of NF-kappa B, a key transcriptional activator of HIV gene expression, in response to UV, suggesting that NF-kappa B acts independently of p38 MAP kinase. In this study, we have investigated whether activation of HIV gene expression occurs when p38 MAP kinase and NF-kappa B are activated by separate stress-causing treatments, each relatively specific for activating only one of the factors. Treatment of cells with sorbitol (hyperosmotic shock) strongly activates p38 MAP kinase, whereas the cytokine TNF alpha is a poor activator of p38 MAP kinase. On the other hand, TNF-alpha is a strong activator of NF-kappa B whereas sorbitol is not. Sorbitol, however, activates AP-1 DNA binding activity in a manner similar to that of UV. Most importantly, both sorbitol and TNF-alpha are poor activators of HIV gene expression in HeLa cells stably transfected with an HIVcat reporter gene, whereas UV elicits a strong response. The combined treatment with UV and hyperosmotic shock produces an additive effect on HIV gene expression, suggesting that these agents activate at least in part by different mechanisms. The combined treatment with sorbitol and TNF-alpha activates p38 and NF-kappa B to levels similar to those with UV, yet only results in 25-30% of the CAT levels elicited by UV. Inhibition of NF-kappa B activation by the protease inhibitor N-alpha-tosyl-L phenylalanine chloromethyl ketone (TPCK) prevents UV activation of HIV gene expression, but does not inhibit p38 MAP kinase activation. We conclude that whereas both p38 MAP kinase and NF-kappa B are important for UV activation of HIV gene expression they act independently from each other and activation of both factors is not sufficient for triggering a full HIV gene expression response. Activation of HIV gene expression by UV must therefore involve additional cellular processes, such as those triggered by DNA damage, for generation of a full gene expression response. PMID- 10677221 TI - Identity of tRNA for yeast tyrosyl-tRNA synthetase: tyrosylation is more sensitive to identity nucleotides than to structural features. AB - The specific aminoacylation of tRNA by yeast tyrosyl-tRNA synthetase does not rely on the presence of modified residues in tRNA(Tyr), although such residues stabilize its structure. Thus, the major tyrosine identity determinants were searched by the in vitro approach using unmodified transcripts produced by T7 RNA polymerase. On the basis of the tyrosylation efficiency of tRNA variants, the strongest determinants are base pair C1-G72 and discriminator residue A73 (the 5' phosphoryl group on C1, however, is unimportant for tyrosylation). The three anticodon bases G34, U35, and A36 contribute also to the tyrosine identity, but to a lesser extent, with G34 having the most pronounced effect. Mutation of the GUA tyrosine anticodon into a CAU methionine anticodon, however, leads to a loss of tyrosylation efficiency similar to that obtained after mutation of the C1-G72 or A73 determinants. Transplantation of the six determinants into four different tRNA frameworks and activity assays on heterologous Escherichia coli and Methanococcus jannaschii tRNA(Tyr) confirmed the completeness of the tyrosine set and the eukaryotic character of the C1-G72 base pair. On the other hand, it was found that tyrosine identity in yeast does not rely on fine architectural features of the tRNA, in particular the size and sequence of the D-loop. Noticeable, yeast TyrRS efficiently charges a variant of E. coli tRNA(Tyr) with a large extra-region provided its G1-C72 base pair is changed to a C1-G72 base pair. Finally, tyrosylation activity is compatible with a +1 shift of the anticodon in the 3'-direction but is strongly inhibited if this shift occurs in the opposite 5'-direction. PMID- 10677220 TI - Catalytic and DNA binding properties of the ogg1 protein of Saccharomyces cerevisiae: comparison between the wild type and the K241R and K241Q active-site mutant proteins. AB - The Ogg1 protein of Saccharomyces cerevisiae belongs to a family of DNA glycosylases and apurinic/apyrimidinic site (AP) lyases, the signature of which is the alpha-helix-hairpin-alpha-helix-Gly/Pro-Asp (HhH-GPD) active site motif together with a conserved catalytic lysine residue, to which we refer as the HhH GPD/K family. In the yeast Ogg1 protein, yOgg1, the HhH-GPD/K motif spans residues 225-260 and the conserved lysine is K241. In this study, we have purified the K241R and K241Q mutant proteins and compared their catalytic and DNA binding properties to that of the wild-type yOgg1. The results show that the K241R mutation greatly impairs both the DNA glycosylase and the AP lyase activities of yOgg1. Specificity constants for cleavage of a 34mer oligodeoxyribonucleotide containing a 7,8-dihydro-8-oxoguanine (8-OxoG) paired with a cytosine, [8-OxoG.C], are 56 x 10(-)(3) and 5 x 10(-)(3) min(-)(1) nM( )(1) for the wild-type and the K241R protein, respectively. On the other hand, the K241Q mutation abolishes the DNA glycosylase and AP lyase activities of yOgg1. In contrast, the K241R and K241Q proteins have conserved wild-type DNA binding properties. K(dapp) values for binding of [8-OxoG.C] are 6.9, 7.4, and 4.8 nM for the wild-type, K241R, and K241Q proteins, respectively. The results also show that AP site analogues such as 1, 3-propanediol (Pr), tetrahydrofuran (F), or cyclopentanol (Cy) are not substrates but constitute good inhibitors of the wild-type yOgg1. Therefore, we have used a 59mer [Pr.C] duplex to further analyze the DNA binding properties of the wild-type, K241R, and K241Q proteins. Hydroxyl radical footprints of the wild-type yOgg1 show strong protection of six nucleotides centered around the Pr lesion in the damaged strand. On the complementary strand, only the cytosine placed opposite Pr was strongly protected. The same footprints were observed with the K241R and K241Q proteins, confirming their wild-type DNA binding properties. These results indicate that the K241Q mutant protein can be used to study interactions between yOgg1 and DNA containing metabolizable substrates such as 8-OxoG or an AP site. PMID- 10677222 TI - Intact aminoacyl-tRNA is required to trigger GTP hydrolysis by elongation factor Tu on the ribosome. AB - GTP hydrolysis by elongation factor Tu (EF-Tu) on the ribosome is induced by codon recognition. The mechanism by which a signal is transmitted from the site of codon-anticodon interaction in the decoding center of the 30S ribosomal subunit to the site of EF-Tu binding on the 50S subunit is not known. Here we examine the role of the tRNA in this process. We have used two RNA fragments, one which contains the anticodon and D hairpin domains (ACD oligomer) derived from tRNA(Phe) and the second which comprises the acceptor stem and T hairpin domains derived from tRNA(Ala) (AST oligomer) that aminoacylates with alanine and forms a ternary complex with EF-Tu. GTP. While the ACD oligomer and the ternary complex containing the Ala-AST oligomer interact with the 30S and 50S A site, respectively, no rapid GTP hydrolysis was observed when both were bound simultaneously. The presence of paromomycin, an aminoglycoside antibiotic that binds to the decoding site and stabilizes codon-anticodon interaction in unfavorable coding situations, did not increase the rate of GTP hydrolysis. These results suggest that codon recognition as such is not sufficient for GTPase activation and that an intact tRNA molecule is required for transmitting the signal created by codon recognition to EF-Tu. PMID- 10677223 TI - The anticodon-binding domain of tyrosyl-tRNA synthetase: state of folding and origin of the crystallographic disorder. AB - The C-terminal domain (residues 320-419) of tyrosyl-tRNA synthetase (TyrRS) from Bacillus stearothermophilus is disordered in the crystal structure. Its function consists of binding the anticodon of tRNA(Tyr). We undertook to characterize its conformational state. A hybrid between the C-terminal fragment and a His-tag sequence was constructed and purified in large amounts. Analyses by mass spectrometry and analytical ultracentrifugation showed that the C-terminal fragment, thus purified, was not degraded and that it neither dimerized nor aggregated. Its far- and near-UV circular dichroism spectra revealed a high content in secondary structures and an asymmetrical environment of its aromatic residues. Each spectrum could be reconstructed by the difference between the corresponding spectra for the full-length TyrRS and for its N-terminal fragment. The Stokes radius of the C-terminal fragment, measured by size exclusion chromatography, indicated a condensed globular state. The fluorescence of ANS (a small hydrophobic probe) showed that the surface of the C-terminal fragment was more hydrophilic than that of a molten globule. These results on the C-terminal fragment and our previous observations that it can undergo cooperative transitions, demonstrated the following points: it is not in a disordered or molten globular state, it has a defined and stable three-dimensional structure, its structures are similar in its isolated and integrated forms, and the apparent disorder in the crystals of the full-length synthetase must be due to the flexibility of the polypeptide segment that links the N- and C-terminal domains. Thus, TyrRS has not evolved strong noncovalent interactions between its catalytic and anticodon-binding domains, contrary to the other synthetases. PMID- 10677224 TI - Squalene synthase: steady-state, pre-steady-state, and isotope-trapping studies. AB - Squalene synthase catalyzes two consecutive reactions in sterol biosynthesis-the condensation of two molecules of farnesyl diphosphate (FPP) to form the cyclopropylcarbinyl intermediate presqualene diphosphate (PSPP) and the subsequent rearrangement and reduction of PSPP to form squalene. Steady-state and pre-steady-state kinetic studies, in combination with isotope-trapping experiments of enzyme.substrate complexes, indicate that two molecules of FPP add to the enzyme before NADPH and that PSPP is converted directly to squalene without dissociating from the enzyme under normal catalytic conditions. In addition, formation of PSPP or a prior conformational change in squalene synthase is the rate-limiting step for synthesis of squalene from FPP via PSPP in the presence of NADPH and for synthesis of PSPP in the absence of NADPH. Squalene synthase is inhibited at high concentrations of FPP. Inhibition is specific for the formation of squalene, but not PSPP, and is competitive with respect to NADPH. In addition, the binding of either NADPH or a third, nonreacting molecule of FPP stimulates the rate of PSPP formation. A kinetic mechanism is proposed to account for these observations. PMID- 10677225 TI - Identification of the sites of hydroxyl radical reaction with peptides by hydrogen/deuterium exchange: prevalence of reactions with the side chains. AB - Hydroxyl radical-effected protium/deuterium ((1)H/(2)H) exchange into the C-H bonds present in peptides has been used to identify the site of hydrogen atom abstraction by hydroxyl radical. Radiolysis of anaerobic, N(2)O-saturated D(2)O solutions containing peptide and dithiothreitol generates a hydroxyl radical that mediates (1)H/(2)H exchange into the side chains of peptides of up to 66 atom % excess (2)H. The (1)H/(2)H exchange is determined by measuring the isotope ratio, [M + H + 1](+)/[M + H](+), of the peptide using electrospray ionization-mass spectrometry. The (1)H/(2)H exchange within each residue of the peptide was determined by measuring the isotope ratio of each isolated dansyl amino acid following hydrolysis and derivatization. Generation of 0.40 mM hydroxyl radical effected (1)H/(2)H exchange into each of the five different residues of (Ala(2)) leucine enkephalin (YAGFL). The propensity of the residues to undergo exchange was L > Y > A congruent with F > G, independent of whether they were radiolyzed separately or as the peptide. The minimal exchange into glycine suggests that reaction of hydroxyl radical with the side chain hydrogens predominates over reaction with the polypeptide alpha-hydrogens. The ability of radiolysis to effect (1)H/(2)H exchange into a larger peptide, SNEQKACKVLGI, was also demonstrated. PMID- 10677226 TI - Drug selectivity is determined by coupling across the NAD+ site of IMP dehydrogenase. AB - Drug resistance often results from mutations that are located far from the drug binding site. The effects of these mutations are perplexing. The inhibition of IMPDH by MPA is an example of this phenomenon. Mycophenolic acid (MPA) is a species-specific inhibitor of IMPDH; mammalian IMPDHs are very sensitive to MPA, while the microbial enzymes are resistant to the inhibitor. MPA traps the covalent intermediate E-XMP and binds in the nicotinamide half of the dinucleotide site. Previous results indicated that about half of the difference in sensitivity derives from residues in the MPA-binding site [Digits, J. A., and Hedstrom, L. (1999) Biochemistry 38, 15388-15397]. The remainder must be attributed to regions outside the MPA-binding site. The adenosine subsite of the NAD+ site is not conserved among IMPDHs and is, therefore, a likely candidate. Our goal is to examine the coupling between the nicotinamide and adenosine sites in order to test this hypothesis. We performed multiple inhibitor experiments with the Tritrichomonas foetus and human type 2 IMPDHs using tiazofurin and ADP, which bind in the nicotinamide and adenosine subsites, respectively. For T. foetus IMPDH, tiazofurin and ADP are extraordinarily synergistic. In contrast, these inhibitors are virtually independent for the human type 2 enzyme. We suggest that the difference in coupling of the nicotinamide and adenosine subsites accounts for the remaining difference in MPA affinity between T. foetus and human IMPDH. PMID- 10677227 TI - The mechanism of orotidine 5'-monophosphate decarboxylase: catalysis by destabilization of the substrate. AB - The mechanism of orotidine 5'-monophosphate decarboxylase (OMP decarboxylase, ODCase) was studied using the decarboxylation of orotic acid analogues as a model system. The rate of decarboxylation of 1,3-dimethylorotic acid and its analogues as well as the stability of their corresponding carbanion intermediates was determined. The results have shown that the stability of the carbanion intermediate is not a critical factor in the rate of decarboxylation. On the other hand, the reaction rate is largely dependent on the equilibrium constant for the formation of a zwitterion. Based on these results, we have proposed a new mechanism in which ODCase catalyzes the decarboxylation of OMP by binding the substrate in a zwitterionic form and providing a destabilizing environment for the carboxylate group of OMP. PMID- 10677228 TI - Kinetic studies of dimeric Ncd: evidence that Ncd is not processive. AB - Ncd is a kinesin-related motor protein which drives movement to the minus-end of microtubules. The kinetics of Ncd were investigated using the dimeric construct MC1 (Leu(209)-Lys(700)) expressed in Escherichia coli strain BL21(DE) as a nonfusion protein [Chandra, R., Salmon, E. D., Erickson, H. P., Lockhart, A., and Endow, S. A. (1993) J. Biol. Chem. 268, 9005-9013]. Acid chemical quench flow methods were used to measure directly the rate of ATP hydrolysis, and stopped flow kinetic methods were used to determine the kinetics of mantATP binding, mantADP release, dissociation of MC1 from the microtubule, and binding of MC1 to the microtubule. The results define a minimal kinetic mechanism, M.N + ATP M.N.ATP M.N.ADP.P N. ADP.P N.ADP + P M.N.ADP M.N + ADP, where N, M, and P represent Ncd, microtubules, and inorganic phosphate respectively, with k(+1) = 2.3 microM(-1) s(-1), k(+2) =23 s(-1), k(+3) =13 s(-1), k(+5)= 0.7 microM(-)(1) s(-)(1), and k(+6) = 3.7 s(-)(1). Phosphate release (k(+4)) was not measured directly although it is assumed to be fast relative to ADP release because Ncd is purified with ADP tightly bound at the active site. ATP hydrolysis occurs at 23 s(-)(1) prior to Ncd dissociation at 13 s(-)(1). The pathway for ATP-promoted detachment (steps 1-3) of Ncd from the microtubule is comparable to kinesin's. However, there are two major differences between the mechanisms of Ncd and kinesin. In contrast to kinesin, mantADP release for Ncd at 3.7 s(-)(1) is the slowest step in the pathway and is believed to limit steady-state turnover. Additionally, the burst amplitude observed in the pre-steady-state acid quench experiments is stoichiometric, indicating that Ncd, in contrast to kinesin, is not processive for ATP hydrolysis. PMID- 10677229 TI - Structural transition at actin's N-terminus in the actomyosin cross-bridge cycle. AB - Force and motion generation by actomyosin involves the cyclic formation and transition between weakly and strongly bound complexes of these proteins. Actin's N-terminus is believed to play a greater role in the formation of the weakly bound actomyosin states than in the formation of the strongly bound actomyosin states. It has been the goal of this project to determine whether the interaction of actin's N-terminus with myosin changes upon transition between these two states. To this end, a yeast actin mutant, Cys-1, was constructed by the insertion of a cysteine residue at actin's N-terminus and replacement of the C terminal cysteine with alanine. The N-terminal cysteine was labeled stoichiometrically with pyrene maleimide, and the properties of the modified mutant actin were examined prior to spectroscopic measurements. Among these properties, actin polymerization, strong S1 binding, and the activation of S1 ATPase by pyrenyl-Cys-1 actin were not significantly different from those of wild type yeast actin, while small changes were observed in the weak S1 binding and the in vitro motility of actin filaments. Fluorescence changes upon binding of S1 to pyrenyl-Cys-1 actin were measured for the strongly (with or without ADP) and weakly (with ATP and ATPgammaS) bound acto-S1 states. The fluorescence increased in each case, but the increase was greater (by about 75%) in the presence of MgATP and MgATPgammaS than in the rigor state. This demonstrates a transition at the S1 contact with actin's N-terminus between the weakly and strongly bound states, and implies either a closer proximity of the pyrene probe on Cys-1 to structural elements on S1 (most likely the loop of residues 626-647) or greater S1-induced changes at the N-terminus of actin in the weakly bound acto-S1 states. PMID- 10677230 TI - Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits. AB - Members of the phospholipase C-beta (PLC-beta) family of proteins are activated either by G alpha or G beta gamma subunits of heterotrimeric G proteins. To define specific regions of PLC-beta 3 that are involved in binding and activation by G beta gamma, a series of fragments of PLC-beta 3 as glutathione-S-transferase (GST) fusion proteins were produced. A fragment encompassing the N-terminal pleckstrin homology (PH) domain and downstream sequence (GST-N) bound to G protein beta 1 gamma 2 in an in vitro binding assay, and binding was inhibited by G protein alpha subunit, G alpha i1. This PLC-beta 3 fragment also inhibited G beta gamma-stimulated PLC-beta activity in a reconstitution system, while having no significant effect on G alpha q-stimulated PLC-beta 3 activity. The N-terminal G beta gamma binding region was delineated further to the first 180 amino acids, and the sequence Asn150-Ser180, just distal to the PH domain, was found to be required for the interaction. Mutation of basic residues 154Arg, 155Lys, 159Lys, and 161Lys to Glu within this region reduced G beta gamma binding affinity and specifically reduced the EC50 for G beta gamma-dependent activation of the mutant enzyme 3-fold. Basal activity and G alpha q-dependent activation of the enzyme were unaffected by the mutations. While these basic residues may not directly mediate the interaction with G beta gamma, the data provide evidence for an N terminal G beta gamma binding region of PLC-beta 3 that is involved in activation of the enzyme. PMID- 10677231 TI - Bovine NAD+-dependent isocitrate dehydrogenase: alternative splicing and tissue dependent expression of subunit 1. AB - NAD+-dependent isocitrate dehydrogenase (IDH), a key regulatory enzyme in the Krebs cycle, is a multi-tetrameric enzyme. At least three of the subunits in the core tetramer of mammals are unique gene products. Subunits 1/beta and 2/gamma are considered to be regulatory, while subunits 3,4/alpha, comprising half the tetramer, are catalytic. The full sequence was obtained for the major subunit 1 cDNA in bovine heart, IDH 1-A. A second cDNA, rare in heart, was also identified (IDH 1-B). Differences in the two were confined to the 3'-region, suggesting alternative splicing. Screening of brain, kidney, and liver RNA showed the presence of IDH 1-A and 1-B and a third major species, IDH 1-C. Amplification of bovine genomic DNA by PCR across the regions of difference produced a single product. Comparison of the genomic and mRNA sequences showed that IDH 1-A resulted from splicing of exon W to exon Y, eliminating intron w, exon X, and intron x. IDH 1-B was formed by splice junctions between exon W, exon X, and exon Y. IDH 1-C resulted from splicing of exon W to exon X and subsequent retention of intron x. The 2 proteins predicted from these 3 mRNAs are identical over their first 357 residues. Protein IDH 1-A, resulting from a termination codon within exon Y, contains an additional 26 residues. Proteins IDH 1-B and 1-C derive from a common termination codon within exon X and contain an additional 28 residues. The two C-terminal regions differ notably in the number and nature of charged residues, resulting in proteins with a charge difference of 3.2 at pH 7.0. Subunit 1 sequences previously reported from other species grouped with one or the other of the bovine proteins. No evidence was found for alternative splicing in subunit 3,4/alpha. The results of the present study, together with recent work on the 2/gamma subunit [Brenner,V., Nyakatura, G., Rosenthal, A., and Platzer, M. (1998) Genomics 44, 8], indicate that the regulatory subunits of the enzyme, but not the catalytic, possess alternatively spliced forms varying in C-terminal properties with tissue-specific expression. The finding is suggestive of a mechanism for modulation of allosteric regulation tailored to the needs of different tissues. PMID- 10677232 TI - Sodium binding site of factor Xa: role of sodium in the prothrombinase complex. AB - The nature of residue 225 on a consensus loop in serine proteases determines whether a protease can bind Na(+). Serine proteases with a Pro at this position are unable to bind Na(+), but those with a Tyr or Phe can bind Na(+). Factor Xa (FXa), the serine protease of the prothrombinase complex, contains a Tyr at this position. Na(+) is also known to stimulate the amidolytic activity of FXa toward cleavage of small synthetic substrates, but the role of Na(+) in the prothrombinase complex has not been investigated. In this study, we engineered a Gla-domainless form of FX (GDFX) in which residue Tyr(225) was replaced with a Pro. We found that Na(+) stimulated the cleavage rate of chromogenic substrates by FXa or GDFXa approximately 8-24-fold with apparent dissociation constants [K(d(app))] of 37 and 182 mM in the presence and absence of Ca(2+), respectively. In contrast, Na(+) minimally affected the cleavage rate of these substrates by the mutant, and no K(d(app)) for Na(+) binding to the mutant could be estimated. Unlike the wild-type enzyme, the reactivity of the mutant with antithrombin was independent of Na(+) and impaired approximately 32-fold. Ca(2+) improved the reactivity of the mutant with antithrombin approximately 5-fold. Affinity of the mutant for binding to factor Va was weakened and its ability to activate prothrombin was severely impaired. Further studies with the wild-type prothrombinase complex revealed that FXa binds to factor Va with a similar K(d(app)) of 1. 1-1.8 nM in the presence of Na(+), K(+), Li(+), Ch(+), and Tris(+) and that the catalytic efficiency of prothrombinase is enhanced less than 1.5-fold by the specific effect of Na(+) in the reaction buffer. These results suggest that (1) the loop including residue 225 (225-loop) is a Na(+) binding site in FXa, (2) the Na(+)- and Ca(2+)-binding loops of FXa are allosterically linked, and (3) the Tyr conformer of the 225-loop is critical for factor Xa function; however, both Na(+)-bound and Na(+)-free forms of factor Xa in the prothrombinase complex can efficiently activate prothrombin. PMID- 10677233 TI - The polar region consecutive to the HIV fusion peptide participates in membrane fusion. AB - The fusion peptide of HIV-1 gp41 is formed by the 16 N-terminal residues of the protein. This 16-amino acid peptide, in common with several other viral fusion peptides, caused a reduction in the bilayer to hexagonal phase transition temperature of dipalmitoleoylphosphatidylethanolamine (T(H)), suggesting its ability to promote negative curvature in membranes. Surprisingly, an elongated peptide corresponding to the 33 N-terminal amino acids raised T(H), although it was more potent than the 16-amino acid fusion peptide in inducing lipid mixing with large unilamellar liposomes of 1:1:1 dioleoylphosphatidylethanolamine/dioleoylphosphatidylcholine/choleste rol. The 17 amino acid C-terminal fragment of the peptide can induce membrane fusion by itself, if it is anchored to a membrane by palmitoylation of the amino terminus, indicating that the additional 17 hydrophilic amino acids contribute to the fusogenic potency of the peptide. This is not solely a consequence of the palmitoylation, as a random peptide with the same amino acid composition with a palmitoyl anchor was less potent in promoting membrane fusion and palmitic acid itself had no fusogenic activity. The 16-amino acid N-terminal fusion peptide and the longer 33-amino acid peptide were labeled with NBD. Fluorescence binding studies indicate that both peptides bind to the membrane with similar affinities, indicating that the increased fusogenic activity of the longer peptide was not a consequence of a greater extent of membrane partitioning. We also determined the secondary structure of the peptides using FTIR spectroscopy. We find that the amino-terminal fusion peptide is inserted into the membrane as a beta-sheet and the 17 C-terminal amino acids lie on the surface of the membrane, adopting an alpha-helical conformation. It was further demonstrated with the use of rhodamine labeled peptides that the 33-amino acid peptide self-associated in the membrane while the 16-amino acid N-terminal peptide did not. Thus, the 16-amino acid N terminal fusion peptide of HIV inserts into the membrane and, like other viral fusion peptides, lowers T(H). In addition, the 17 consecutive amino acids enhance the fusogenic activity of the fusion peptide presumably by promoting its self association. PMID- 10677234 TI - Changes of intrinsic membrane potentials induced by flip-flop of long-chain fatty acids. AB - The passive transbilayer movement-flip-flop-was investigated on planar bilayer lipid membranes (BLMs), containing myristic, stearic, or linoleic long-chain fatty acids (FA). In response to a transbilayer pH gradient, a difference in the surface charges between inner and outer leaflets appeared. Because the BLM was formed from FA and neutral lipid, a surface potential difference was originated solely by a concentration difference of the initially equally distributed ionized FA. As revealed by zeta-potential measurements, the corresponding surface potential difference DeltaPhi(s) was at least twice the value expected from a titration of the FA alone. The additional surface charge was attributed to FA flip-flop induced by the transbilayer pH gradient. DeltaPhi(s) was derived from capacitive current measurements carried out with a direct current (dc) bias and was corrected for changes of membrane dipole potential Phi(d). Dual-wavelength ratiometric fluorescence measurements have shown that Phi(d) values of the pure DPhPC bilayers and BLMs containing 40 mol % FA differ by less than 6%. It is concluded that fast FA flip-flop is not restricted to membranes with high curvature. The role of pH gradient as an effective driving force for the regulation of FA uptake is discussed. PMID- 10677235 TI - Mg2+ activates 5-lipoxygenase in vitro: dependency on concentrations of phosphatidylcholine and arachidonic acid. AB - Mg2+ gave dose-dependent activation of 5-lipoxygenase (5LO) in vitro. As for Ca2+, the activation depended on the presence of phosphatidylcholine (PC) vesicles, and the activation response was different at various combinations of arachidonate and PC. Stimulation of 5LO activity was observed with Mg2+ concentrations of 0.1-1 mM, similar to the concentration range of free Mg2+ in mammalian cells. However, to observe a clear increase in 5LO hydrophobicity, a higher concentration of Mg2+ (4 mM) was required, and at this concentration also 5LO activation was optimal. Combinations of Mg2+ with ATP (containing free Mg2+ and MgATP2- complex) gave better activation of 5LO than either agent alone. This effect of Mg2+ (and ATP) could be of interest in relation to basal 5LO activity in cells not subjected to a particular stimulus. PMID- 10677236 TI - Alpha-ketoacids are potent slow binding inhibitors of the hepatitis C virus NS3 protease. AB - The replication of the hepatitis C virus (HCV), an important human pathogen, crucially depends on the proteolytic maturation of a large viral polyprotein precursor. The viral nonstructural protein 3 (NS3) harbors a serine protease domain that plays a pivotal role in this process, being responsible for four out of the five cleavage events that occur in the nonstructural region of the HCV polyprotein. We here show that hexapeptide, tetrapeptide, and tripeptide alpha ketoacids are potent, slow binding inhibitors of this enzyme. Their mechanism of inhibition involves the rapid formation of a noncovalent collision complex in a diffusion-limited, electrostatically driven association reaction followed by a slow isomerization step resulting in a very tight complex. pH dependence experiments point to the protonated catalytic His 57 as an important determinant for formation of the collision complex. K(i) values of the collision complexes vary between 3 nM and 18.5 microM and largely depend on contacts made by the peptide moiety of the inhibitors. Site-directed mutagenesis indicates that Lys 136 selectively participates in stabilization of the tight complex but not of the collision complex. A significant solvent isotope effect on the isomerization rate constant is suggestive of a chemical step being rate limiting for tight complex formation. The potency of these compounds is dominated by their slow dissociation rate constants, leading to complex half-lives of 11-48 h and overall K(i) values between 10 pM and 67 nM. The rate constants describing the formation and the dissociation of the tight complex are relatively independent of the peptide moiety and appear to predominantly reflect the intrinsic chemical reactivity of the ketoacid function. PMID- 10677237 TI - Pressure-induced unfolding/refolding of ribonuclease A: static and kinetic Fourier transform infrared spectroscopy study. AB - In this paper, we illustrate the use of high-pressure Fourier transform infrared (FT-IR) spectroscopy to study the reversible presssure-induced unfolding and refolding of ribonuclease A (RNase A) and compare it with the results obtained for the temperature-induced transition. FT-IR spectroscopy monitors changes in the secondary structural properties (amide I' band) or tertiary contacts (tyrosine band) of the protein upon pressurization or depressurization. Analysis of the amide I' spectral components reveals that the pressure-induced denaturation process sets in at 5. 5 kbar at 20 degrees C and pH 2.5. It is accompanied by an increase in disordered structures while the content of beta sheets and alpha-helices drastically decreases. The denatured state above 7 kbar retains nonetheless some degree of beta-like secondary structure and the molecule cannot be described as an extended random coil. Increase of pH from 2.5 to 5.5 has no influence on the structure of the pressure-denatured state; it slightly changes the stability of the protein only. All experimental evidence indicates that the pressure-denatured states of monomeric proteins have more secondary structure than the temperature-denatured states. Different modes of denaturation, including pressure, may correlate differently with the roughness of the energy scale and slope of the folding funnel. For these reasons we have also carried out pressure-jump kinetic studies of the secondary structural evolution in the unfolding/refolding reaction of RNase A. In agreement with the theoretical model presented by Hummer et al. [(1998) Proc. Natl. Acad. Sci. U.S.A. 95, 1552-1555], the experimental data show that pressure slows down folding and unfolding kinetics (here 1-2 orders of magnitude), corresponding to an increasingly rough landscape. The kinetics remains non-two-state under pressure. Assuming a two-step folding scenario, the calculated relaxation times for unfolding of RNase A at 20 degrees C and pH 2.5 can be estimated to be tau(1) approximately 0.7 min and tau(2) approximately 17 min. The refolding process is considerably faster (tau(1) approximately 0.3 min, tau(2) approximately 4 min). Our data show that the pressure stability and pressure-induced unfolding/refolding kinetics of monomeric proteins, such as wild-type staphylococcal nuclease (WT SNase) and RNase A, may be significantly different. The differences are largely due to the four disulfide bonds in RNase A, which stabilize adjacent structures. They probably lead to the much higher denaturation pressure compared to SNase, and this might also explain why the volume change of WT SNase upon unfolding is about twice as large. PMID- 10677238 TI - Expression and membrane assembly of a transmembrane region from Neu. AB - Transmembrane domains of receptor tyrosine kinases are increasingly seen as key modulatory elements in signaling pathways. The present work addresses problems surrounding expression, isolation, secondary structure recovery, and assembly into membranes, of the relatively large quantities of transmembrane peptides needed to investigate these pathways by NMR spectroscopy. We demonstrate significant correspondence between SDS-PAGE behavior of such peptides and their (2)H NMR spectra in lipid bilayer membranes. A 50-residue peptide, Neu(exp), containing the transmembrane portion of the receptor tyrosine kinase, Neu, was designed for expression in Escherichia coli. The sequence also contained 11-12 amino acids from each side of the transmembrane domain. The common problem of low expressivity of transmembrane peptides was encountered-likely associated with membrane toxicity of the desired gene product. This difficulty was overcome by expressing the peptide as a TrpE fusion protein in a pATH vector to target expression products to inclusion bodies, and subsequently removing the TrpE portion by cyanogen bromide cleavage. Inclusion bodies offered the additional benefits of reduced proteolytic degradation and simplified purification. The presence of a hexa-His tag allowed excellent recovery of the final peptide, while permitting use of denaturing solvents and avoiding the need for HPLC with its attendant adsorption losses. Isolated expressed peptides were found to be pure, but existed as high oligomers rich in beta-structure as evidenced by CD spectroscopy and SDS-PAGE behavior. Dissolution in certain acidic organic solvents led to material with increased alpha-helix content, which behaved in detergent as mixtures of predominantly monomers and dimers-a situation often considered to exist in cell membranes. For purposes of NMR spectroscopy, peptide alanine residues were deuterated in high yield during expression. The same acidic organic solvents used to dissolve and dissociate expressed transmembrane peptides proved invaluable for their assembly into lipid bilayers. Analogous transmembrane peptides from the human receptor tyrosine kinase, ErbB-2, demonstrated related phenomena. PMID- 10677239 TI - Nanosecond dynamics of tryptophans in different conformational states of apomyoglobin proteins. AB - Fluorescence anisotropy kinetics were employed to quantify the nanosecond mobility of tryptophan residues in different conformational states (native, molten globule, unfolded) of apomyoglobins. Of particular interest is the similarity between the fluorescence anisotropy decays of tryptophans in the native and molten globule states. We find that, in these compact states, tryptophan residues rotate rapidly within a cone of semiangle 22-25 degrees and a correlation time of 0.5 ns, in addition to rotating together with the whole protein with a correlation time of 7-11 ns. The similar nanosecond dynamics of tryptophan residues in both states suggests that the conformation changes that distinguish the molten globule and native states of apomyoglobins originate from either subtle, slow rearrangements or fast changes distant from these tryptophans. PMID- 10677240 TI - Copper-induced conformational changes in the N-terminal domain of the Wilson disease copper-transporting ATPase. AB - The Wilson disease copper-transporting ATPase plays a critical role in the intracellular trafficking of copper. Mutations in this protein lead to the accumulation of a toxic level of copper in the liver, kidney, and brain followed by extensive tissue damage and death. The ATPase has a novel amino-terminal domain ( approximately 70 kDa) which contains six repeats of the copper binding motif GMTCXXC. We have expressed and characterized this domain with respect to the copper binding sites and the conformational consequences of copper binding. A detailed analysis of this domain by X-ray absorption spectroscopy (XAS) has revealed that each binding site ligates copper in the +1 oxidation state using two cysteine side chains with distorted linear geometry. Analysis of copper induced conformational changes in the amino-terminal domain indicates that both secondary and tertiary structure changes take place upon copper binding. These copper-induced conformational changes could play an important role in the function and regulation of the ATPase in vivo. In addition to providing important insights on copper binding to the protein, these results suggest a possible mechanism of copper trafficking by the Wilson disease ATPase. PMID- 10677241 TI - Treatment of experimental autoimmune encephalomyelitis with adenylate deaminase from Penicillium lanoso-viride. AB - The effect of intramuscularly administered immunomodulator, adenylate deaminase (E.C. 3.5.4.6), from Penicillium lanoso-viride on the clinical score of acute experimental autoimmune encephalomyelitis (EAE), a T cell-mediated autoimmune disease, was examined by inoculation of guinea pigs with rabbit brain and spinal cord homogenate (encephalitogen) and complete Freund's adjuvant. Adenylate deaminase (ADA) was effective in delaying the onset of clinical disease. ADA inhibited the severity of EAE. There was a significant decrease in clinical signs. A decrease in the number of morbid and dead animals was observed. Of ADA treated animals, 50-80% developed no clinical manifestations of EAE. The optimal version of treatment was a single preventive injection of ADA 1 day before the sensitization and then every second day after immunization for 20 days. ADA treatment of immunized animals diminished the activity of 2', 3'-cyclic nucleotide 3'-phosphodiesterase in the cerebrospinal fluid, as well the amount of complement fixing antiencephalitogenic antibodies in the blood serum. The mechanism of ADA cerebroprotective action is discussed. Significant skin-allergic cross-reaction of delayed-type hypersensitivity between ADA and encephalitogen was observed. PMID- 10677243 TI - Cytotoxic T lymphocyte antigen 4 (CD152) regulates self-reactive T cells in BALB/c but not in the autoimmune NOD mouse. AB - Recent studies demonstrated that engagement of cytotoxic T lymphocyte antigen 4 (CTLA-4)/(CD152) generates an inhibitory signal to T cells which arrests an on going immune response. Since aberrant CD152 activity is thought to contribute to autoimmunity, we examined the effect of CD152-mediated inhibitory signals on the response to self and foreign antigens in autoimmune, diabetes-prone NOD and non autoimmune BALB/c mice. The interaction of CD152 with its ligand B7 was prevented by treating the mice with anti-CD152 blocking antibody, before and following immunization of the mice with the self-antigen, syngeneic islet cells, or the foreign antigen, key-hole limpet hemocyanin (KLH). CD152 blockade in BALB/c mice stimulated a robust islet-specific T cell-mediated immune response compared to control antibody-treated mice. The augmentation of T cell responses in BALB/c mice was consistent with the role proposed for CD152 as a down-regulator of T cell activation responses. Furthermore, CD152 blockade unmasked islet cell specific autoreactive T cells in the non-autoimmune BALB/c mouse. Conversely, CD152 blockade in NOD mice failed to regulate islet-specific auto-reactive T cell responses. However, CD152 blockade enhanced the T cell response to the exogenous, foreign antigen KLH in both non-autoimmune BALB/c and autoimmune NOD mice. Collectively, these results suggest that there is not a global defect in CD152 mediated regulation of peripheral T cell immune responses in NOD autoimmune mice but rather, a defect specific to T cells recognizing self antigen. PMID- 10677242 TI - Immunization of low-density lipoprotein receptor deficient (LDL-RD) mice with heat shock protein 65 (HSP-65) promotes early atherosclerosis. AB - Heat shock proteins are a family of approximately 25 highly conserved proteins upregulated in response to various forms of stress. They play an active role in the development autoimmune diseases in animals, and have been incriminated in human autoimmune diseases (i.e. rheumatoid arthritis, multiple sclerosis). It has been previously shown, that an induced immune response against Heat shock protein 65 (HSP-65) results in atherosclerotic lesions in normocholesterolemic rabbits. We have supported these findings showing that C57BL/6 mice immunized with HSP-65 and fed a high-fat diet develop enhanced fatty streaks. To create a model that will eliminate the need for exogenous supplementation of a high-fat diet, we have immunized LDL receptor deficient (LDL-RD) mice with HSP-65 or with heat-killed Mycobacterium tuberculosis (Mt). Seven groups of LDL-RD mice (n=10), were immunized subcutaneously with different concentrations of HSP-65, Mt or bovine serum albumin (BSA). All mice were fed a normal chow-diet for 3 months. The mice immunized with the higher doses of Mt developed significantly larger fatty streaks when compared with their BSA immunized littermates. The size of the lesions in the aortic sinus were: 31,562+/-5,994 microm(2)in the 10 microg Mt and 52,777+/-5,245 microm(2)in the 100 microg Mt vs. 11, 500+/-3,750 microm(2)in the BSA group (P<0.05). In the HSP-65-immunized mice, only the group injected with the highest dose (5 microg, twice) developed significantly larger fatty streaks when compared with the BSA-immunized group (28,611+/-4,716 microm(2)vs. 11,500+/ 3,750 microm(2)respectively, (P<0.05). The HSP-65-but not the Mt- or BSA immunized mice developed high titers of anti HSP-65 antibodies, beginning 10 days after the immunization, which persisted until they were killed. Immunohistochemical staining showed CD3-positive lymphocytes in the aortic sinus of mice immunized with Mt or HSP-65, but not in the control group. Thus, we established a mouse model of HSP-65 immune mediated atherosclerosis devoid of high fat diet supplementation. This model will enable us to further study the role of the immune system in atherosclerosis, via HSP-65 and raise novel immunomodulatory therapeutic modalities. PMID- 10677244 TI - Prevention of diabetes in the NOD mouse by a Th1 clone specific for a hsp60 peptide. AB - Peptide-based therapies have been shown to be effective in the prevention of diabetes in the NOD mouse. We have been interested in the T cell response elicited by such therapies and have been studying a T cell clone (C3.5) specific for hsp60 AA 437-460, generated following immunization with the hsp60 437-460 peptide. The C3.5 clone was CD4(+), Vbeta8.3 TCR(+), I-A(g7)restricted and of the Th1 type. The injection of this clone into prediabetic NOD mice prevented the adoptive transfer of the disease and suppressed the development of spontaneous diabetes. This effect was reflected in a reduction in the degree and severity of insulitis in mice injected with this clone. In addition, an antibody response was elicited to the C3.5 clone in mice given multiple injections of the clone. The epitope recognized by C3.5 is located in the N-terminus of the hsp60 AA 437-460 peptide, and this clone was unable to recognize the native hsp60 molecule. These data raise questions concerning the mechanism by which peptide-based therapies prevent autoimmune disease. PMID- 10677245 TI - Endothelial injury in internal organs of University of California at Davis line 200 (UCD 200) chickens, an animal model for systemic sclerosis (Scleroderma). AB - Systemic sclerosis (SSc) is a multisystem disorder characterized by mononuclear cell infiltration and fibrosis. Using skin samples from human SSc and UCD 200 chickens, which spontaneously develop a hereditary disease closely resembling human SSc, we have shown previously that endothelial cell apoptosis is a primary event in the pathogenesis of SSc. The aim of the present study was to investigate the initial disease stage in visceral organs of UCD 200 chickens with special emphasis on endothelial apoptosis, mononuclear cell infiltration and collagen deposition using tissue samples from oesophagus, lung, heart, kidney and liver. Apoptotic endothelial cells were detected by terminal deoxynucleotidyl transferase-mediated FITC-dUTP nick end labeling (TUNEL), mononuclear cell infiltrates were stained with hematoxylin and eosin, and increased collagen deposition was demonstrated by Goldner staining. Apoptotic endothelial cells were detected in oesophagus, lung and kidney of UCD 200 chickens at the initial stage of the disease. No apoptotic endothelial cells were found in heart or liver of UCD 200 or in visceral organs of healthy normal UCD 058 control chickens. Oesophagus of UCD 200 chickens, which was the most affected internal organ, showed mononuclear cell infiltrations and increased deposition of collagen. Perivascular inflammatory infiltrates and collagen deposition appeared later than endothelial cell apoptosis. These data support the hypothesis that endothelial cell apoptosis initiates the disease process, followed by mononuclear cell infiltration and fibrosis. PMID- 10677246 TI - Treatment of lupus in NZB/W F1 mice with monoclonal antibody against Fas ligand. AB - Since Fas ligand (FasL) can induce apoptosis of Fas-bearing cells, Fas/FasL interactions can play a critical role in maintaining self-tolerance. Fas/FasL interactions in lupus-like autoimmune disease have been well characterized in studies using either Fas or FasL mutant mice. However, the effect of the defective FasL-mediated signaling on the establishment of lupus in other mouse strains, such as NZB/W (B/W) F1, remains uncertain. In the present study, we examined the effect of anti-FasL monoclonal antibody (mAb) on the development of lupus. Treatment of B/W F1 mice with anti-FasL mAb augmented IgG1- and IgG2a-type anti-dsDNA Ab production. However, treatment of B/W F1 mice with anti-FasL mAb also significantly prevented the development of lupus nephritis. These results indicate that Fas/FasL interactions not only regulate IgG-type autoantibody production, but also influence the development of lupus nephritis in B/W F1 mice. PMID- 10677247 TI - Histone-containing immune complexes are to a large extent responsible for anti dsDNA reactivity in the Farr assay of active SLE patients. AB - Increased titres of anti-dsDNA antibodies, especially if of high avidity, are associated with renal exacerbations in patients with systemic lupus erythematosus (SLE). One of the most reliable assays to measure anti-dsDNA antibodies, the Farr assay, is believed to detect preferentially high avidity antibodies. Purified non complexed monoclonal antibodies (mAbs) against nucleosomes, obtained from mice with SLE, are not reactive in the Farr assay, but can become so once complexed to nucleosomes. These Farr-positive, nucleosome containing, immune complexes were also able to bind in vivo to the glomerular basement membrane (GBM), predominantly via heparan sulphate (HS). To evaluate whether in SLE patients the same kind of immune complexes are responsible for Farr reactivity, IgG from serum or plasma was isolated under dissociating and physiological conditions. We observed that after purification under dissociating conditions, Farr reactivity was significantly decreased (P<0.0001) in contrast to reactivity with histones and two 'control' antigens: Epstein Barr Virus (EBV) and Ro/SS-A. Reactivity with nucleosomes also decreased after purification, although to a lesser extent. Plasma purified under physiological conditions showed no decrease in Farr reactivity. The importance of histones for the generation of immune complexes is supported by the two following observations. Firstly, the presence of histones could be demonstrated in serum and plasma of SLE patients but not in serum of healthy controls or in IgG preparations purified under dissociating conditions. Secondly, Farr reactivity of purified IgG preparations could be restored by addition of purified histones. From these studies we conclude that histones containing immune complexes are responsible for a large part of the Farr reactivity in active SLE, and are therefore indirectly implicated in the pathogenesis of lupus nephritis. PMID- 10677248 TI - A susceptibility locus for human systemic lupus erythematosus (hSLE1) on chromosome 2q. AB - To identify chromosomal regions containing susceptibility loci for systemic lupus erythematosus (SLE), we performed genome scans in families with multiple SLE patients from Iceland, a geographical and genetic isolate, and from Sweden. A number of chromosomal regions showed maximum lod scores (Z) indicating possible linkage to SLE in both the Icelandic and Swedish families. In the Icelandic families, five regions showed lod scores greater than 2.0, three of which (4p15 13, Z=3.20; 9p22, Z=2.27; 19q13, Z=2.06) are homologous to the murine regions containing the lmb2, sle2 and sle3 loci, respectively. The fourth region is located on 19p13 (D19S247, Z=2.58) and the fifth on 2q37 (D2S125, Z=2.06). Only two regions showed lod scores above 2.0 in the Swedish families: on chromosome 2q11 (D2S436, Z=2. 13) and 2q37 (D2S125, Z=2.18). The combination of both family sets gave a highly significant lod score at D2S125 of Z=4.24 in favor of linkage for 2q37. This region represents a new locus for SLE. Our results underscore the importance of studying well-defined populations for genetic analysis of complex diseases such as SLE. PMID- 10677249 TI - Human autoantibodies to a novel Golgi protein golgin-67: high similarity with golgin-95/gm 130 autoantigen. AB - Autoantibodies to subcellular organelles have been described in patients with various systemic rheumatic diseases and our laboratories have been focused on studies of the Golgi complex as the autoimmune target. We have previously isolated and described four of the five known Golgi autoantigens reported to date. During the characterization of Golgi autoantigen golgin-95/gm130, another human cDNA that shared a significant degree of similarity in both nucleotide and amino acid sequences was identified. Analysis of cDNAs from different libraries suggested that this is a distinct gene encoding a protein of 67 kDa which has four regions with sequence identity to gm130, ranging between 42 and 60%. In this report, we describe the complete cDNA encoding a closely related Golgi protein provisionally named golgin-67. Among a group of 84 human anti-Golgi sera, five (6%) were shown to recognize golgin-67. Anti-golgin-67 human sera and affinity purified rabbit antibody to the recombinant protein gave predominant Golgi staining. Golgin-67 is thus the smallest member of a growing family of Golgi autoantigens rich in alpha-helical coiled-coil domain. The current hypothesis for the generation of autoimmune antibody to the Golgi complex is discussed. PMID- 10677250 TI - Latent autoimmune thyroiditis in untreated patients with HCV chronic hepatitis: a case-control study. AB - In order to establish a relationship between Hepatitis C virus (HCV) chronic infection and autoimmune thyroiditis, 97 untreated patients with biopsy-proven HCV chronic hepatitis and 97 controls were studied. An ultrasound examination of the thyroid and an assay of serum thyroid-stimulating hormone (TSH), thyroid hormones and anti-thyroid antibodies were performed in all cases. The overall prevalence of thyroid abnormalities was higher in patients than in controls (17 vs. 4%, P<0.01) and the prevalence of anti-thyroid antibodies was significantly different between the two groups (P<0. 02). HCV patients with (n=13) compared to HCV patients without anti-thyroid antibodies (n=84) were older, predominantly female, and more frequently had increased serum TSH levels or a hypoechogenic pattern of the thyroid gland, while Knodell's score and prevalence of cirrhosis were similar. Latent autoimmune thyroiditis is more frequent in untreated HCV patients than in controls. This finding raises questions about the mechanism of autoimmunity induced by HCV and provides an explanation for the high rate of overt autoimmune thyroiditis during interferon treatment in these patients. PMID- 10677251 TI - Bilaterian origins: significance of new experimental observations. AB - Several recent laboratory observations that bear on the origin of the Bilateria are reviewed and interpreted in light of our set-aside cell theory for bilaterian origins. We first discuss new data concerning the phylogeny of bilaterian phyla. Next, we use systematic, molecular, and paleontological lines of evidence to argue that the latest common ancestor of echinoderms plus hemichordates used a maximal indirect mode of development. Furthermore, the latest common ancestor of molluscs and annelids was also indirectly developing. Finally, we discuss new data on Hox gene expression patterns which suggest that both sea urchins and polychaete annelids use Hox genes in a very similar fashion. Neither utilizes the complete Hox complex in the development of the larva per se, while the Hox complex is expressed in the set-aside cells from which the adult body plan is formed. Our current views on the ancestry of the bilaterians are summarized in phylogenetic terms, incorporating the characters discussed in this paper. PMID- 10677252 TI - Mesenchyme with fgf-10 expression is responsible for regenerative capacity in Xenopus limb buds. AB - A young tadpole of an anuran amphibian can completely regenerate an amputated limb, and it exhibits an ontogenetic decline in the ability to regenerate its limbs. However, whether mesenchymal or epidermal tissue is responsible for this decrease of the capacity remains unclear. Moreover, little is known about the molecular interactions between these two tissues during regeneration. The results of this study showed that fgf-10 expression in the limb mesenchymal cells clearly corresponds to the regenerative capacity and that fgf-10 and fgf-8 are synergistically reexpressed in regenerating blastemas. However, neither fgf-10 nor fgf-8 is reexpressed after amputation of a nonregenerative limb. Nevertheless, nonregenerative epidermal tissue can reexpress fgf-8 under the influence of regenerative mesenchyme, as was demonstrated by experiments using a recombinant limb composed of regenerative limb mesenchyme and nonregenerative limb epidermis. Taken together, our data demonstrate that the regenerative capacity depends on mesenchymal tissue and suggest that fgf-10 is likely to be involved in this capacity. PMID- 10677253 TI - Sacral neural crest cells colonise aganglionic hindgut in vivo but fail to compensate for lack of enteric ganglia. AB - The vagal neural crest is the origin of majority of neurons and glia that constitute the enteric nervous system, the intrinsic innervation of the gut. We have recently confirmed that a second region of the neuraxis, the sacral neural crest, also contributes to the enteric neuronal and glial populations of both the myenteric and the submucosal plexuses in the chick, caudal to the level of the umbilicus. Results from this previous study showed that sacral neural crest derived precursors colonised the gut in significant numbers only 4 days after vagal-derived cells had completed their migration along the entire length of the gut. This observation suggested that in order to migrate into the hindgut and differentiate into enteric neurons and glia, sacral neural crest cells may require an interaction with vagal-derived cells or with factors or signalling molecules released by them or their progeny. This interdependence may also explain the inability of sacral neural crest cells to compensate for the lack of ganglia in the terminal hindgut of Hirschsprung's disease in humans or aganglionic megacolon in animals. To investigate the possible interrelationship between sacral and vagal-derived neural crest cells within the hindgut, we mapped the contribution of various vagal neural crest regions to the gut and then ablated appropriate sections of chick vagal neural crest to interrupt the migration of enteric nervous system precursor cells and thus create an aganglionic hindgut model in vivo. In these same ablated animals, the sacral level neural axis was removed and replaced with the equivalent tissue from quail embryos, thus enabling us to document, using cell-specific antibodies, the migration and differentiation of sacral crest-derived cells. Results showed that the vagal neural crest contributed precursors to the enteric nervous system in a regionalised manner. When quail-chick grafts of the neural tube adjacent to somites 1-2 were performed, neural crest cells were found in enteric ganglia throughout the preumbilical gut. These cells were most numerous in the esophagus, sparse in the preumbilical intestine, and absent in the postumbilical gut. When similar grafts adjacent to somites 3-5 or 3-6 were carried out, crest cells were found within enteric ganglia along the entire gut, from the proximal esophagus to the distal colon. Vagal neural crest grafts adjacent to somites 6-7 showed that crest cells from this region were distributed along a caudal-rostral gradient, being most numerous in the hindgut, less so in the intestine, and absent in the proximal foregut. In order to generate aneural hindgut in vivo, it was necessary to ablate the vagal neural crest adjacent to somites 3-6, prior to the 13-somite stage of development. When such ablations were performed, the hindgut, and in some cases also the cecal region, lacked enteric ganglionated plexuses. Sacral neural crest grafting in these vagal neural crest ablated chicks showed that sacral cells migrated along normal, previously described hindgut pathways and formed isolated ganglia containing neurons and glia at the levels of the presumptive myenteric and submucosal plexuses. Comparison between vagal neural crest-ablated and nonablated control animals demonstrated that sacral-derived cells migrated into the gut and differentiated into neurons in higher numbers in the ablated animals than in controls. However, the increase in numbers of sacral neural crest-derived neurons within the hindgut did not appear to be sufficiently high to compensate for the lack of vagal-derived enteric plexuses, as ganglia containing sacral neural crest-derived neurons and glia were small and infrequent. Our findings suggest that the neuronal fate of a relatively fixed subpopulation of sacral neural crest cells may be predetermined as these cells neither require the presence of vagal-derived enteric precursors in order to colonise the hindgut, nor are capable of dramatically altering their proliferation or d PMID- 10677254 TI - The Ets domain transcription factor Erm distinguishes rat satellite glia from Schwann cells and is regulated in satellite cells by neuregulin signaling. AB - Distinct glial cell types of the vertebrate peripheral nervous system (PNS) are derived from the neural crest. Here we show that the expression of the Ets domain transcription factor Erm distinguishes satellite glia from Schwann cells beginning early in rat PNS development. In developing dorsal root ganglia (DRG), Erm is present both in presumptive satellite glia and in neurons. In contrast, Erm is not detectable at any developmental stage in Schwann cells in peripheral nerves. In addition, Erm is downregulated in DRG-derived glia adopting Schwann cell traits in culture. Thus, Erm is the first described transcription factor expressed in satellite glia but not in Schwann cells. In culture, the Neuregulin1 (NRG1) isoform GGF2 maintains Erm expression in presumptive satellite cells and reinduces Erm expression in DRG-derived glia but not in Schwann cells from sciatic nerve. These data demonstrate that there are intrinsic differences between these glial subtypes in their response to NRG1 signaling. In neural crest cultures, Erm-positive progenitor cells give rise to two distinct glial subtypes: Erm-positive, Oct-6-negative satellite glia in response to GGF2, and Erm negative, Oct-6-positive Schwann cells in the presence of serum and the adenylate cyclase activator forskolin. Thus, Erm-positive neural crest-derived progenitor cells and presumptive satellite glia are able to acquire Schwann cell features. Given the in vivo expression of Erm in peripheral ganglia, we suggest that ganglionic Erm-positive cells may be precursors of Schwann cells. PMID- 10677255 TI - Anterior endoderm is sufficient to rescue foregut apoptosis and heart tube morphogenesis in an embryo lacking retinoic acid. AB - The vitamin A deficient (VAD) quail embryo lacks active retinoids, fails to express normally GATA-4, and develops a nonlooping heart tube morphogenetic defect that is a model for congenital cardiomyopathy. VAD quail embryos, or chick embryos depleted specifically for GATA factors, show in addition abnormal foregut development, characterized by apoptosis of the endoderm cells associated with presumptive myocardium during the process of heart tube formation. Exogenous retinoic acid or transplantation of normal chick embryo anterior endoderm is sufficient to rescue apoptosis as well as GATA-4 expression and results in normal development and heart tube morphogenesis. Normal posterior endoderm also contains retinoids but is unable to rescue the VAD defect. Our results indicate that a retinoid-dependent transcriptional program, mediated at least in part by GATA factors, is critical in presumptive foregut endoderm for normal heart tube morphogenesis. PMID- 10677256 TI - Smad5 is essential for left-right asymmetry in mice. AB - Left-right (L-R) asymmetry of the vertebrate body plan is established from an originally morphologically symmetric embryo. Recent studies have implicated several TGF-beta family signaling proteins (i.e., nodal, lefty-1, lefty-2, activin receptor type IIB, and Smad2) in L-R axis determination in the mouse. However, the genetic pathways underlying L-R patterning are still unclear. Smad5 is a downstream component in the TGF-beta family signaling cascade, and lack of Smad5 results in embryonic lethality between E9.5 and E11.5. In this report, we demonstrate that Smad5 mutant embryos have defects in heart looping and embryonic turning which are the first signs of L-R asymmetry in mice. To gain more insights into the molecular basis of the laterality defects in the Smad5-deficient embryos, we examined the expression of lefty-1, lefty-2, nodal, and Pitx2 since the asymmetric expression of these genes always closely correlates with the direction of heart looping and embryonic turning. In the absence of Smad5, lefty 1 was expressed at very low or undetectable levels, while nodal, lefty-2, and Pitx2 were expressed bilaterally. These data suggest that Smad5 is upstream of lefty-1, nodal, and lefty-2, and as a consequence also of Pitx2, and Smad5 is essential for L-R axis determination. PMID- 10677257 TI - Sacral neural crest cell migration to the gut is dependent upon the migratory environment and not cell-autonomous migratory properties. AB - Avian neural crest cells from the vagal (somite level 1-7) and the sacral (somite level 28 and posterior) axial levels migrate into the gut and differentiate into the neurons and glial cells of the enteric nervous system. Neural crest cells that emigrate from the cervical and thoracic levels stop short of the dorsal mesentery and do not enter the gut. In this study we tested the hypothesis that neural crest cells derived from the sacral level have cell-autonomous migratory properties that allow them to reach and invade the gut mesenchyme. We heterotopically grafted neural crest cells from the sacral axial level to the thoracic level and vice versa and observed that the neural crest cells behaved according to their new position, rather than their site of origin. Our results show that the environment at the sacral level is sufficient to allow neural crest cells from other axial levels to enter the mesentery and gut mesenchyme. Our study further suggests that at least two environmental conditions at the sacral level enhance ventral migration. First, sacral neural crest cells take a ventral rather than a medial-to-lateral path through the somites and consequently arrive near the gut mesenchyme many hours earlier than their counterparts at the thoracic level. Our experimental evidence reveals only a narrow window of opportunity to invade the mesenchyme of the mesentery and the gut, so that earlier arrival assures the sacral neural crest of gaining access to the gut. Second, the gut endoderm is more dorsally situated at the sacral level than at the thoracic level. Thus, sacral neural crest cells take a more direct path to the gut than the thoracic neural crest, and also their target is closer to the site from which they initiate migration. In addition, there appears to be a barrier to migration at the thoracic level that prevents neural crest cells at that axial level from migrating ventral to the dorsal aorta and into the mesentery, which is the portal to the gut. PMID- 10677258 TI - beta-catenin in epithelial morphogenesis: conversion of part of avian foot scales into feather buds with a mutated beta-catenin. AB - We explored the role of beta-catenin in chicken skin morphogenesis. Initially beta-catenin mRNA was expressed at homogeneous levels in the epithelia over a skin appendage tract field which became transformed into a periodic pattern corresponding to individual primordia. The importance of periodic patterning was shown in scaleless mutants, in which beta-catenin was initially expressed normally, but failed to make a punctuated pattern. To test beta-catenin function, a truncated armadillo fragment was expressed in developing chicken skin from the RCAS retrovirus. This produced a variety of phenotypic changes during epithelial appendage morphogenesis. In apteric and scale-producing regions, new feather buds with normal-appearing follicle sheaths, dermal papillae, and barb ridges were induced. In feather tracts, short, wide, and curled feather buds with abnormal morphology and random orientation formed. Epidermal invaginations and placode like structures formed in the scale epidermis. PCNA staining and the distribution of molecular markers (SHH, NCAM, Tenascin-C) were characteristic of feather buds. These results suggest that the beta-catenin pathway is involved in modulating epithelial morphogenesis and that increased beta-catenin pathway activity can increase the activity of skin appendage phenotypes. Analogies between regulated and deregulated new growths are discussed. PMID- 10677259 TI - Hydra metalloproteinase 1: a secreted astacin metalloproteinase whose apical axis expression is differentially regulated during head regeneration. AB - The newly emerging astacin metalloproteinase family comprises multiple members with diverse functions. Most recently, the development-related functions have been attributed to both (1) proteolytic cleavage and subsequent release of active TGF-beta-like growth factors from latent inhibitory complexes and (2) modification of extracellular matrix (ECM) assembly and composition. We previously identified and purified hydra metalloproteinase 1 (HMP-1), a developmentally important astacin proteinase that functions in head regeneration and transdifferentiation of tentacle battery cells (L. Yan et al., 1995, Development 121, 1591-1602). In the present study, further cloning revealed that HMP-1 is produced as a secreted zymogen with a conserved hydrophobic signal sequence and a putative propeptide. The processed HMP-1 is composed of a characteristic astacin proteinase domain and a unique Cys-rich C-terminus. With this simple domain structure, HMP-1 represents an ancestral astacin proteinase. Consistent with its role in head regeneration, HMP-1 mRNA is expressed at highest levels by endodermal cells at the apical pole of the body column just inferior to the base of tentacles, the region of active cell differentiation or transdifferentiation. A modified immunocytochemical procedure demonstrated that HMP-1 protein can be localized not only to ECM of tentacles as we previously reported, but also to endodermal cells of the body column in a pattern similar to its mRNA distribution. The localization of HMP-1 protein in tentacles was confirmed using an enzymatic approach. A translocation of HMP-1 protein from cells in the body column to the extracellular milieu in tentacles further suggests that HMP-1 is a secreted protein. HMP-1 expression undergoes extensive regulation at the transcriptional level both temporally and spatially during head regeneration. The involvement of HMP-1 in this morphogenetic process is further supported by the blockage of head regeneration with localized antisense treatment. Taken together, these results suggest that HMP-1 is a secreted astacin metalloproteinase that has an important role in regulating hydra head morphogenesis potentially through its differential expression along the body axis. PMID- 10677260 TI - Sequential programs of retinoic acid synthesis in the myocardial and epicardial layers of the developing avian heart. AB - Endogenous patterns of retinoic acid (RA) signaling in avian cardiac morphogenesis were characterized by localized expression of a key RA-synthetic enzyme, RALDH2, which displayed a biphasic pattern during heart development. RALDH2 immunoreactivity was initially apparent posterior to Hensen's node of stage 5-6 embryos and subsequently in somites and unsegmented paraxial and lateral plate mesoderm overlapping atrial precursors in the cardiogenic plate of stage 9- embryos. Initial RALDH2 synthesis in the posterior myocardium coincided with activation of the AMHC1 gene, a RA-responsive marker of inflow heart segments. A wave of RALDH2 synthesis then swept the myocardium in a posterior-to anterior direction, reaching the outflow tract by stage 13, then fading from the myocardial layer. The second phase of RALDH2 expression, initiated at stage 18 in the proepicardial organ, persisted in migratory epicardial cells that completely enveloped the heart by stage 24. Early restriction of RALDH2 expression to the posterior cardiogenic plate, overlapping RA-inducible gene activation, provides evidence for commitment of posterior avian heart segments by localized production of RA, whereas subsequent RALDH2 expression exclusively in the migratory epicardium suggests a role for the morphogen in ventricular expansion and morphogenesis of underlying myocardial tissues. PMID- 10677261 TI - The HMG box transcription factor gene Sox14 marks a novel subset of ventral interneurons and is regulated by sonic hedgehog. AB - Cell-type diversity along the dorsoventral axis of the developing neural tube is influenced by factors secreted by groups of cells at the dorsal and ventral midline. Upon reception of these signals, precursor cells express specific sets of transcription factors which, in turn, play critical roles in cell-type specification. Here we report the cloning and characterization of Sox14, a novel and highly conserved member of the Sry-related Sox transcription factor gene family, in mouse and chick. Sox14 expression is restricted to a limited population of neurons in the developing brain and spinal cord of both species. Sox14 marks a subset of interneurons at a defined dorsoventral position adjacent to ventral motor neurons in the spinal cord. In vivo grafting of chick notochord tissue to ectopic positions adjacent to the developing spinal cord altered the expression domain of Sox14. Furthermore, expression of Sox14 in spinal cord explants was found to be regulated by Sonic hedgehog in a dose-dependent manner. These data implicate a novel class of transcription factors in dorsoventral cell type specification in the spinal cord. PMID- 10677262 TI - A 130-kDa membrane protein of sperm flagella is the receptor for asterosaps, sperm-activating peptides of starfish Asterias amurensis. AB - Spermatozoa of the starfish, Asterias amurensis, have a specific receptor for asterosap, a sperm-activating peptide isolated from the jelly coat of homologous eggs. We characterized the receptor by using several asterosap derivatives. Analysis of equilibrium binding of radioactive di-iodinated Bolton-Hunter reagent labeled asterosap ((125)I(2)-BHP15) to the spermatozoa indicated that the cell has 1.1 x 10(5) binding sites of high affinity (K(d) = 57 pM), and also the receptor showed positive cooperativity for asterosap binding. When spermatozoa were treated with fluorophore-labeled asterosap, the sperm flagella were labeled, indicating that the receptors are mostly localized in the sperm tail. When spermatozoa were reacted with radioactive asterosap prelabeled with photoaffinity cross-linkers, a single 130-kDa membrane protein of sperm flagella was specifically radiolabeled. This result was reproducible regardless of the length of spacer arm of cross-linkers so far studied. Therefore, the 130-kDa protein is likely to be the receptor for asterosaps. Modification of asterosap at the N terminal region with bulky molecules such as carboxyfluorescein did not affect the activity of asterosap, suggesting that the N-terminus of asterosap is not involved in the ligand-receptor interaction. On the other hand, S-alkylated asterosaps did not compete with (125)I(2)-BHP15 for binding to the receptor, indicating that disulfide linkage of asterosap is essential for the ligand receptor interaction. The properties of the receptor, high affinity and high concentration, enabled us to apply the fluorescence polarization technique to study the molecular interaction between asterosap and the receptor. Using this method, we performed binding experiments in almost real time and found that divalent cations are significantly involved in the interaction between asterosap and the receptor. PMID- 10677263 TI - Development of methods to assess the effects of xenobiotics in outdoor artificial streams. AB - Two collaborative research projects were designed to develop and validate methods for determining the chronic effects of xenobiotics in freshwater ecosystems. The work reported here focuses on the development of methods for measuring effects on fish, invertebrates, and algae in outdoor artificial streams. 3,4-Dichloroaniline (3,4-DCA), has been used as a reference xenobiotic in two artificial stream experiments. The first used five stream channels: a control and treatments ranging from 70 to 2400 microg/liter. The second used eight stream channels--a control and treatments ranging from 0.45 to 4700 microg/liter and four coupled, 510-liter-capacity, downstream ponds-a control and treatments of 1.7, 37, and 820 microg/liter. Effects on the biota of the stream channels and the downstream ponds were examined using a range of sampling techniques and in situ toxicity tests. PMID- 10677264 TI - In vitro cytotoxicity of aromatic aerobic biotransformation products in bluegill sunfish BF-2 cells. AB - Toluene (methylbenzene) is a common environmental pollutant that is found in many hazardous waste sites and it is an aquifer contaminant. A concern is the potential risk to human and ecosystem health due to exposure to toluene and its major biotransformation products. The cytotoxicity of eight aromatic products of toluene aerobic biotransformation was investigated in bluegill sunfish BF-2 cells. The cytotoxicity was determined using several in vitro assay endpoints. BF 2 cells were propagated at 32 degrees C in an atmosphere of 5% CO2-95% air. The concentrations of these products causing 50% inhibition in cell replication, protein content, uptake of natural red, and colony formation were evaluated and compared. The results of the study indicate a direct relationship between the exposure concentration of these products and observed cytotoxic effects. In descending order of cytotoxicity, the compounds were 3-methylcatechol, 4 methylcatechol, catechol, o-cresol, p-cresol, m-cresol, benzaldehyde, and methyl benzoate. PMID- 10677265 TI - Influence of nitrogen status on the bioconcentration of hydrophobic organic compounds to Selenastrum capricornutum. AB - Changes in algal nitrogen status that increase algal lipid content also affect the bioconcentration of hydrophobic organic compounds (HOCs). Bioconcentration factors (BCFs) for several HOCs increased up to nine times as the total algal lipid content of the green algae Selenastrum carpricornutum increased from 17 to 44% of the algal dry weight as a consequence of nitrogen starvation. An increase in total lipid from 17 to 44% should theoretically increase the BCFs by a factor of 2.6. BCFs for PCB 31, PCB 49, PCB 153, and DDT increased with maximum lipid content by factors of 6.3, 8.9, 8.9, and 6.6, respectively, thus more than theoretically predicted from the lipid normalization of BCFs obtained at exponential growth phase (17% total lipid for S. carpricornutum), whereas BCFs for PCB 105, phenanthrene, and 4-chloroaniline increased at 44% lipid content, only by factors of 1.5, 1.5, and 2.5, respectively, and thus less than or equal to the theoretical prediction. Lipid-class normalization of BCFs did not reveal significant information beyond that available from normalizing to total lipid. PMID- 10677266 TI - Simultaneous geno- and immunotoxicological investigations for early detection of organophosphate toxicity in rats. AB - Detectability of toxic effects by repeated doses of dimethoate (DM) and methylparathion (MPT) were investigated by geno-and immunotoxicological methods in male Wistar rats following a 28-day oral exposure. In the dose range of 28.2, 14.1, and 7.04, and 7.04 mg/kg/day DM, the two higher doses decreased the body weight gain. The top dose increased the weight of liver, kidneys, and testicles; the white blood cell count; and the cell content of the femoral bone marrow. From immune function parameters measured [IgM-plaque forming cells (PFC) assay, delayed-type hypersensitivity (DTH) reaction] only the maximum of the DTH reaction decreased at the top dose. Of the MPT doses (0.872, 0.436, and 0.218 mg/kg/day) the two higher ones increased the liver weight, and a dose-dependent increase was found in the MCV value. No evaluable changes in the examined immune function parameters were observed. Both substances increased the number of numerical but not the structural chromosome aberrations at lower dose levels (the two larger doses of DM, and all the three doses of MPT) than those ones which caused changes in the examined immune function parameters. According to these results, the genotoxicological approach seems to be more sensitive for detection of repeated-dose oral toxicity of the investigated two organophosphates than the immunotoxicological one. PMID- 10677267 TI - Concentration effects of selected insecticides on brain acetylcholinesterase in the common carp (Cyprinus carpio L.). AB - The differential inhibition of acetylcholinesterase (AChE) by organophosphate (OP) and carbamate (C) is followed by the distinct duration of exposure effect on common carp AChE. Hence, in the present study in vivo exposure period effect and in vitro concentration-response of chlorfenvinphos, chlorpyrifos diazinon, and carbofuran were investigated on Cyprinus carpio L. AChE. Individuals of 1-year old carp were exposed for 96 h to different concentrations of insecticides, after which AChE activity was measured in the brain. The highest concentrations of carbofuran (2.44 mg x L(-1)), chlorfenvinphos (2.9 mg x L(-1)), and diazinon (2.5 mg x L(-1)) killed all the test animals after only 4 h, although there was no statistically significant difference from the control group's brain AChE activity. The lowest concentration significantly inhibited brain AChE after 96 h. Chlorfenvinphos was the most potent inhibitor in vivo and chlorpyrifos the least active inhibitor after 96 h of exposure time. In vitro experimentation with the same pesticide indicated that several concentrations inhibited 50% of the AChE activity (I50), ranging from 4.1x10(-7) to 8.12x10(-4) M in both single inhibitory action and joint inhibitory effect. The results suggest that in biomonitoring programs carp brain AChE can be a good diagnostic tool for chronic OP nd C pollution. PMID- 10677268 TI - Simultaneous action of cypermethrin and two environmental pollutant metals, cadmium and lead, on bone marrow cell chromosomes of rats in subchronic administration. AB - The aim of the present study was to investigate the possible genotoxic effects, exerted by the pyrethroid cypermethrin and by either of the metals cadmium and lead alone or in combination, on bone marrow cell chromosomes in a subchronic experiment. Outbred male Wistar rats were treated per os for 4 weeks in a five time per week schedule with 5.54, 11.08, or 22.16 mg/kg cypermethrin (1/100, 1/50, 1/25 LD50) alone, or in combination of 1/100 and 1/50 LD50 cypermethrin with 2.0 mg/kg cadmium chloride or 10 mg/kg lead acetate. On the day following the last treatment, the animals were sacrificed and bone marrow from the femur was prepared. Twenty metaphases from 10 animals per group were evaluated. The evaluation comprised the frequency of aberrant cells, the numerical and structural aberrations, and the alterations in relative organ weights. In the dosage used, cypermethrin and cadmium alone caused no significant increase in the chromosomal aberrations, and lead acetate caused an increase of the numerical aberrations only. Combination of cypermethrin and cadmium also failed to induce significant chromosomal effects. The cypermethrin + lead combination, however, induced a significant increase of structural chromosomal aberrations, predominantly of all acentric fragments. This lead to the conclusion that the simultaneous administration of lead and cypermethrin results in an enhanced genotoxic effect. PMID- 10677269 TI - Application of quantitative structure--activity relationships for assessing the aquatic toxicity of phthalate esters. AB - Phthalate esters (PEs) are an important class of industrial chemicals for which an extensive aquatic toxicity database is available. The objectives of this study were to use these data to develop quantitative structure-activity relationships (QSARs) that describe aquatic toxicity for different freshwater and marine species, gain insights into toxicity mechanisms, and calculate PE water quality criteria using statistical extrapolation procedures. Results for low-molecular weight PEs with log Kow<6 indicate that toxicity data conform to a simple log Kow dependent QSAR. Fish were found to be more sensitive than algae while invertebrates spanned a wide range in toxicological response. Freshwater and marine species demonstrated a similar distribution of sensitivities. Comparison of species-dependent QSARs supports the hypothesis that biotransformation plays an important role in explaining toxicity differences observed between species. Estimated critical body residues (CBRs) for parent PE in fish were in the range reported for other polar organic chemicals while CBRs for parent PE plus associated metabolites were in the range reported for nonpolar narcotics (i.e., baseline toxicity) suggesting a possible putative role of PE metabolites. Depending on extrapolation procedure and assumptions, predicted no-effect concentrations (PNECs) for dimethyl, diethyl, dibutyl, and butybenzyl phthalate ranged from 3109 to 4780, 865 to 1173, 43 to 62, and 38 to 60 microg l(-1), respectively. PNECs derived using this approach provide a transparent technical basis to support aquatic risk assessment for low-molecular-weight PEs. Results for high-molecular-weight PEs (log Kow>6) indicate that these chemicals are not acutely or chronically toxic to freshwater or marine organisms due to the combined role of low water solubility and limited bioconcentration potential which precludes attainment of internal concentrations that are required to elicit adverse effects. It is concluded that attempts to establish aquatic PNECs for high-molecular-weight PEs are not scientifically defensible. PMID- 10677270 TI - Potential of the insecticides acephate and methamidophos to contaminate groundwater. AB - The possible contamination of groundwater by the insecticides acephate and methamidophos was assessed using the behavior assessment model (BAM) and the groundwater pollution-potential model (GWP). The dissipation coefficients of the two insecticides in two soils (Annei silt loam and Pingchen silt clay loam) at different moisture contents (50 and 100% field capacity) and soil temperatures (20 and 30 degrees C) were studied by determining the degradation and adsorption of each insecticide in the soil. The movement of acephate and methamidophos was studied by leaching each insecticide in a soil column in the laboratory. The absorption coefficient of methamidophos was much higher than that of acephate in both types of soil. In the leaching test, methamidophos more easily leached out from the Pingchen soil column than from the Annei soil column. Methamidophos was rapidly degraded, with a half-life of 1.11 to 1.61 days in the Annei soil and 7.50 to 13.20 days in the Pingchen soil at different temperatures and soil water contents. Acephate was found to have a longer half-life than methamidophos in soil; however, the mobility of methamidophos in both soils was slower than that of acephate. The mobility of acephate in soil is somewhat faster than that of methamidophos, and thus acephate may lead to the contamination of groundwater much more easily than methamidophos under normal conditions. PMID- 10677271 TI - Sensitivity and significance of luminescent bacteria in chronic toxicity testing based on growth and bioluminescence. AB - This study explored the use of luminescent bacteria (Vibrio fischeri) for chronic aquatic toxicity tests. The evaluated inhibition of growth to Cu2+, Cr6+, Zn2+, Hg2+, Cd2+, Pb2+, cetyl-trimethylammonium bromide, 3,4-dichloroaniline, acetone, dimethylsulfoxide, ethanol, nitrobenzene, methanol, and 3,5-dichlorophenol was compared with results from another investigation, where the inhibition was determined by bioluminescence. Growth inhibition was found to indicate more reliably the presence of substances with chronic toxic properties than the loss of bioluminescence. But growth responded weaker to the majority of the analyzed toxicants than bioluminescence. This must be connected with the test parameters and the experimental conditions. But among growth experiments with freshwater bacteria species the sensitivity of the growth inhibition assay with V. fischeri is competitive when a poor medium is employed. PMID- 10677272 TI - Environmental Research Section A. PMID- 10677273 TI - Tyrosinase kinetics: a semi-quantitative model of the mechanism of oxidation of monohydric and dihydric phenolic substrates. AB - A mathematical model of phase I melanogenesis is described based on the differential reactivity of tyrosinase according to the redox status of the active site copper atoms shown by Lerch and co-workers (see Lerch, 1981, Metal Ions in Biological Systems (Sigel, H., ed.) Vol. 13, pp. 143-186. New York: Marcel Dekker) in combination with the indirect formation of the catecholic intermediate substrate. In this model the unusual autoactivation kinetics of tyrosinase are explained by recruitment of enzyme from the met -form, in which the active-site copper atoms are in the oxidized (Cu(II)) state, by 2-electron donation from catechol oxidation. Using estimates of the values for the rate constants of the six reactions involved, the general characteristics of the model are shown to be consistent with the kinetic behaviour of tyrosinase in vitro. These include a lag period which is sensitive to catechol addition. PMID- 10677274 TI - General equilibrium of an ecosystem. AB - Ecosystems and economies are inextricably linked: ecosystem models and economic models are not linked. Consequently, using either type of model to design policies for preserving ecosystems or improving economic performance omits important information. Improved policies would follow from a model that links the systems and accounts for the mutual feedbacks by recognizing how key ecosystem variables influence key economic variables, and vice versa. Because general equilibrium economic models already are widely used for policy making, the approach used here is to develop a general equilibrium ecosystem model which captures salient biological functions and which can be integrated with extant economic models. In the ecosystem model, each organism is assumed to be a net energy maximizer that must exert energy to capture biomass from other organisms. The exerted energies are the "prices" that are paid to biomass, and each organism takes the prices as signals over which it has no control. The maximization problem yields the organism's demand for and supply of biomass to other organisms as functions of the prices. The demands and supplies for each biomass are aggregated over all organisms in each species which establishes biomass markets wherein biomass prices are determined. A short-run equilibrium is established when all organisms are maximizing and demand equals supply in every biomass market. If a species exhibits positive (negative) net energy in equilibrium, its population increases (decreases) and a new equilibrium follows. The demand and supply forces in the biomass markets drive each species toward zero stored energy and a long-run equilibrium. Population adjustments are not based on typical Lotka Volterra differential equations in which one entire population adjusts to another entire population thereby masking organism behavior; instead, individual organism behavior is central to population adjustments. Numerical simulations use a marine food web in Alaska to illustrate the model and to show several simultaneous predator/prey relationships, prey switching by the top predator, and energy flows through the web. PMID- 10677275 TI - On optimal size and number of reserves for metapopulation persistence. AB - Habitat fragmentation is generally considered to be detrimental to the persistence of natural populations. In nature management, one therefore tends to prefer few large nature reserves over many small nature reserves having equal total area. This paper examines whether this preference is warranted in a metapopulation framework with circular reserves (patches) by formulating the dependence of metapopulation persistence on the size and number of reserves, both of which depend on reserve radius if the total area is kept constant. Two measures of metapopulation persistence are used: R(0), the number of patches colonized by an occupied patch during its lifetime as an occupied patch, and T(e), the expected time to extinction. These two measures are functions of the extinction and colonization rates of the metapopulation. Several mechanisms for the extinction and colonization processes are formulated from which the dependence of these rates on reserve radius is calculated. It turns out that T(e)generally increases with reserve radius for all mechanisms, which supports the preference of few large reserves. However, R(0)supports this preference only in the case of some special, rather unrealistic, mechanisms. In many other, more realistic, cases an intermediate reserve size exists for which metapopulation persistence measured by R(0)is optimal. PMID- 10677276 TI - Individual-based perspectives on R(0). AB - Without doubt the basic reproductive ratio, R(0), is the most widely used quantity in epidemic theory. Standard compartmental models show how R(0)is related to the average age of infection, vaccination thresholds for eradication and equilibrium solutions. However, many of the basic formulae for R(0)break down when we consider transmission of infection to be a stochastic process involving discrete individuals. This paper clarifies why and when these differences arise and predicts when individual-based considerations are likely to be important in modelling infection dynamics. PMID- 10677277 TI - A fragment of recombinant GABA(A) receptor alpha1 subunit forming rosette-like homo-oligomers. AB - The type A gamma-aminobutyric acid (GABA(A)) receptor plays a major role in inhibitory synaptic transmission in the central nervous system. A fragment consisting of residues Cys166 to Leu296 of the alpha1 subunit of the GABA(A) receptor was overexpressed in Escherichia coli and was found to have stable beta rich structures. Here, results from laser scattering, gel electrophoresis and electron microscopy demonstrated that this recombinant protein formed rosette like homo-oligomers, mainly pentamers in solution. Therefore, the fragment apparently provides a valuable model system for studying the pentameric holoreceptor assembly. Non-reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis of the fragment showed that disulfide bonds formed between monomers contributed to the oligomerization of the fragment. The fact that this fragment alone could form pentamers in vitro strongly suggests that amino acid residues located within the Cys166-Leu296 region of the alpha1 subunit may contribute to the oligomerization of GABA(A) receptor in vivo. PMID- 10677278 TI - Ligand-triggered stabilization of vitamin D receptor/retinoid X receptor heterodimer conformations on DR4-type response elements. AB - Nuclear receptors integrate an incoming signal in the form of a nuclear hormone by undergoing a conformational change that results via co-activator proteins in an activation of the basal transcriptional machinery. The vitamin D(3) receptor is the nuclear receptor for 1alpha,25-dihydroxyvitamin D(3 )(1alpha,25(OH)(2)D(3)) and is known to function as a heterodimer with the retinoid X receptor on DR3-type 1alpha,25(OH)(2)D(3) response elements. Here, it could be demonstrated that DR4-type response elements are at least as effective as DR3-type 1alpha,25(OH)(2)D(3) response elements. Gel shift clipping analysis showed that vitamin D(3) receptor-retinoid X receptor heterodimers form in response to 1alpha, 25(OH)(2)D(3) and retinoid X receptor ligands, the pan agonist 9-cis retinoic acid (9cRA) and the retinoid X receptor-selective retinoid CD2425, different conformations on the DR4-type element of the rat Pit-1 gene. Interestingly, on this response element the heterodimeric complexes of retinoid X receptor with the thyroid hormone receptor, the retinoic acid receptor and the benzoate ester receptor also displayed characteristic individual ligand-dependent complex formation. On the level of complex formation, utilizing DNA affinity and functional assays, only vitamin D(3) receptor-retinoid X receptor heterodimers showed a synergistic interaction of both ligands. However, the sensitivity of vitamin D(3) receptor-retinoid X receptor heterodimers to 1alpha,25(OH)(2)D(3) was found to be much higher than to retinoid X receptor ligands. Taken together, this study demonstrates a unique interaction potential of vitamin D(3) receptor and retinoid X receptor but also establishes DR4-type response elements as multi functional DNA binding sites with a potential to integrate various hormone signalling pathways. PMID- 10677279 TI - IS911 transposition is regulated by protein-protein interactions via a leucine zipper motif. AB - Efficient intermolecular transposition of bacterial insertion sequence IS911 involves the activities of two element-encoded proteins: the transposase, OrfAB, and a regulatory factor, OrfA. OrfA shares the majority of its amino acid sequence with the N-terminal part of OrfAB. This includes a putative helix-turn helix and three of four heptads of a leucine zipper motif. OrfA strongly stimulates OrfAB-mediated intermolecular transposition both in vivo and in vitro. The present results support the notion that this is accomplished by direct interaction between these two proteins via the leucine zipper. We used both a genetic approach, based on gene fusions with phage lambda repressor, and a physical approach, involving co-immunoprecipitation, to show that OrfA not only undergoes oligomerisation but is capable of engaging with OrfAB to form heteromultimers, and that the leucine zipper is necessary for both types of interaction. Furthermore, mutation of the leucine zipper in OrfA inactivated its regulatory function. Previous observations demonstrated that the integrity of the leucine zipper motif was also important for OrfAB binding to the IS911 terminal inverted repeats. Here, we show, in gel shift experiments, using a derivative of OrfAB deleted for the C-terminal catalytic domain, OrfAB[1-149], that the protein is capable of pairing two inverted repeats to generate a species resembling a "synaptic complex". Preincubation of OrfAB[1-149] with OrfA dramatically reduced formation of this complex and favored formation of an alternative complex devoid of OrfA. Together these results suggest that OrfA exerts its regulatory effect by interacting transiently with OrfAB via the leucine zipper and modifying OrfAB binding to the inverted repeats. PMID- 10677280 TI - Mechanisms ensuring rapid and complete DNA replication despite random initiation in Xenopus early embryos. AB - Chromosome replication initiates without sequence specificity at average intervals of approximately 10 kb during the rapid cell cycles of early Xenopus embryos. If the distribution of origins were random, some inter-origin intervals would be too long to be fully replicated before the end of S phase. To investigate what ensures rapid completion of DNA replication, we have examined the replication intermediates of plasmids of various sizes (5.3-42.2 kbp) in Xenopus egg extracts by two-dimensional gel electrophoresis and electron microscopy. We confirm that replication initiates without sequence specificity on all plasmids. We demonstrate for the first time that multiple initiation events occur on large plasmids, but not on small (<10 kb) plasmids, at average intervals of approximately 10 kb. Origin interference may prevent multiple initiation events on small plasmids. Multiple initiation events are neither synchronous nor regularly spaced. Bubble density is higher on later than on earlier replication intermediates, showing that initiation frequency increases throughout S phase, speeding up replication of late intermediates. We suggest that potential origins are abundant and randomly distributed, but that the increase of initiation frequency during S phase, and possibly origin interference, regulate origin activation to ensure rapid completion of replication. PMID- 10677281 TI - Conformational flexibility of B-DNA at 0.74 A resolution: d(CCAGTACTGG)(2). AB - The affinity and specificity of a ligand for its DNA site is a function of the conformational changes between the isolated and complexed states. Although the structures of a hydroxypyrrole-imidazole-pyrrole polyamide dimer with 5' CCAGTACTGG-3' and the trp repressor recognizing the sequence 5'-GTACT-3' are known, the baseline conformation of the DNA site would contribute to our understanding of DNA recognition by these ligands. The 0.74 A resolution structure of a B-DNA double helix, 5'-CCAGTACTGG-3', has been determined by X-ray crystallography. Six of the nine phosphates, two of four bound calcium ions and networks of water molecules hydrating the oligonucleotide have alternate conformations. By contrast, nine of the ten bases have a single, unique conformation with hydrogen atoms visible in most cases. The polyamide molecules alter the geometry of the phosphodiester backbone, and the water molecules mediating contacts in the trp repressor/operator complex are conserved in the unliganded DNA. Furthermore, the multiple conformational states, ions and hydration revealed by this ultrahigh resolution structure of a B-form oligonucleotide are potentially general considerations for understanding DNA binding affinity and specificity by ligands. PMID- 10677282 TI - DNA sequence context modulates the impact of a cisplatin 1,2-d(GpG) intrastrand cross-link on the conformational and thermodynamic properties of duplex DNA. AB - The anticancer activity of cisplatin derives from its ability to bind and cross link DNA, with the major adduct being the 1,2-d(GpG) intrastrand cross-link. Here, the consequences of this adduct on the conformation, thermal stability, and energetics of duplex DNA are assessed, and the modulation of these parameters by the sequence context of the adduct is evaluated. The properties of a family of 15 mer DNA duplexes containing a single 1,2-d(GpG) cis-?Pt(NH(3))(2)?(2+) intrastrand cross-link are probed in different sequence contexts where the flanking base-pairs are systematically varied from T.A to C.G to A.T. By using a combination of spectroscopic and calorimetric techniques, the structural, thermal, and thermodynamic properties of each duplex, both with and without the cross-link, are characterized. Circular dichroism spectroscopic data reveal that the cross-link alters the structure of the host duplex in a manner consistent with a shift from a B-like to an A-like conformation. Thermal denaturation data reveal that the cross-link induces substantial thermal and thermodynamic destabilization of the host duplex. Significantly, the magnitudes of these cross link-induced effects on duplex structure, thermal stability, and energetics are influenced by the bases that flank the adduct. The presence of flanking A.T base pairs, relative to T.A or C.G base-pairs, enhances the extent of cross-link induced alteration to an A-like conformation and dampens the extent of cross-link induced duplex destabilization. These results are discussed in terms of available structural data, and in terms of the selective recognition of cisplatin-DNA adducts by HMG-domain proteins. PMID- 10677283 TI - Three conformations of an archaeal chaperonin, TF55 from Sulfolobus shibatae. AB - Chaperonins are cylindrical, oligomeric complexes, essential for viability and required for the folding of other proteins. The GroE (group I) subfamily, found in eubacteria, mitochondria and chloroplasts, have 7-fold symmetry and provide an enclosed chamber for protein subunit folding. The central cavity is transiently closed by interaction with the co-protein, GroES. The most prominent feature specific to the group II subfamily, found in archaea and in the eukaryotic cytosol, is a long insertion in the substrate-binding region. In the archaeal complex, this forms an extended structure acting as a built-in lid, obviating the need for a GroES-like co-factor. This extension occludes a site known to bind non native polypeptides in GroEL. The site and nature of substrate interaction are not known for the group II subfamily. The atomic structure of the thermosome, an archaeal group II chaperonin, has been determined in a fully closed form, but the entry and exit of protein substrates requires transient opening. Although an open form has been investigated by electron microscopy, conformational changes in group II chaperonins are not well characterized. Using electron cryo-microscopy and three-dimensional reconstruction, we describe three conformations of a group II chaperonin, including an asymmetric, bullet-shaped form, revealing the range of domain movements in this subfamily. PMID- 10677284 TI - Sequential determination of ligands binding to discrete components in heterogeneous mixtures by iterative panning and blocking (IPAB). AB - Biopanning has been used extensively in conjunction with purified components, but there are also examples in which mixtures of targets have been investigated. This study introduces a methodological innovation, termed iterative panning and blocking (IPAB), to extend the range of specific interactions that can be probed in mixtures. Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A series of peptides containing the Gly-Pro-Arg-Pro motif efficiently blocked the binding of phage having the same motif, presumably by binding to their common target. A second round of panning was performed against the same target mixture in the presence of this blocking peptide. Phage recovered from this second panning exhibited a motif (Ser-His-Tyr) that was subsequently shown to bind specifically to complement component C1q. A second peptide containing this motif specifically blocked the interaction of the phage with C1q. A third round of panning performed in the presence of both the fibrinogen- and the C1q- blocking peptides yielded phage with a new peptide motif (Asn-Pro-Phe) that also bound specifically to C1q, apparently at a new site. The three motifs isolated through this iterative process were distinct in that each was blocked only by its corresponding peptide. This IPAB strategy can be applied to many high diversity selection procedures that target complex mixtures. PMID- 10677285 TI - Guided selection of a pan carcinoma specific antibody reveals similar binding characteristics yet structural divergence between the original murine antibody and its human equivalent. AB - Antibody engineering provides an excellent tool for the generation of human immunotherapeutics for the targeted treatment of solid tumours. We have engineered and selected a completely human antibody to epithelial glycoprotein-2 (EGP-2), a transmembrane glycoprotein present on virtually all human simple epithelia and abundantly expressed on a variety of human carcinomas. We chose to use the procedure of "guided selection" to rebuild a high-affinity murine antibody into a human antibody, using two consecutive rounds of variable domain shuffling and phage library selection. As a starting antibody, the murine antibody MOC-31 was used. After the first round of guided selection, where the V(H) of MOC-31 was combined in Fab format with a human V(L)C(L) library, a small panel of human light chains was identified, originating from a segment of the VkappaIII family, whereas the MOC-31 V(L) is more homologous to the VkappaII family. Nevertheless, one of the chimaeric Fabs, C3, displayed an off-rate similar to MOC-31 scFv. Combining the V(L) of C3 with a human V(H) library, while retaining the V(H) CDR3 of MOC-31, clones were selected using human V(H) genes originating from the rarely used V(H)7 family. The best clone, 9E, shows over 13 amino acid mutations from the germline sequence, has an off-rate comparable to the original antibody and specifically binds to the "MOC-31"-epitope on EGP-2 in specificity and competition ELISA, FACS analysis and immunohistochemistry. In both V(L) and V(H) of antibody 9E, three germline mutations were found creating the MOC-31 homologue residue. Structural modelling of both murine and human antibodies reveals that one of the germline mutations, 53Y in V(H) CDR2, is likely to be involved in antigen binding. We conclude that, although they may bind the same epitope and have similar binding affinity to the antigen as the original murine antibody, human antibodies derived by guided selection unlike CDR grafted antibodies, may retain only some of the original key elements of the binding site chemistry. The selected human anti-EGP-2 antibody will be a suitable reagent for tumour targeting. PMID- 10677286 TI - Solution structure of an 11-mer duplex containing the 3, N(4)-ethenocytosine adduct opposite 2'-deoxycytidine: implications for the recognition of exocyclic lesions by DNA glycosylases. AB - Lipid peroxidation products, as well as the metabolic products of vinyl chloride, react with cellular DNA producing the mutagenic adduct 3,N(4)-etheno-2' deoxycytidine (epsilondC), along with several other exocyclic derivatives. High resolution NMR spectroscopy and restrained molecular dynamics simulations were used to establish the solution structure of an 11-mer duplex containing an epsilondC.dC base-pair at its center. The NMR data suggested a regular right handed helical structure having all residues in the anti orientation around the glycosydic torsion angle and Watson-Crick alignments for all canonical base-pairs of the duplex. Restrained molecular dynamics generated a three-dimensional model in excellent agreement with the spectroscopic data. The (epsilondC. dC)-duplex structure is a regular right-handed helix with a slight bend at the lesion site and no severe distortions of the sugar-phosphate backbone. The epsilondC adduct and its partner dC were displaced towards opposite grooves of the helix, resulting in a lesion-containing base-pair that was highly sheared but stabilized to some degree by the formation of a single hydrogen bond. Such a sheared base pair alignment at the lesion site was previously observed for epsilondC.dG and epsilondC.T duplexes, and was also present in the crystal structures of duplexes containing dG.T and dG. U mismatches. These observations suggest the existence of a substrate structural motif that may be recognized by specific DNA glycosylases during the process of base excision repair. PMID- 10677287 TI - Projection structure of a transcriptional regulator, HupR, determined by electron cryo-microscopy. AB - Large, well-ordered two-dimensional crystals of the histidine-tagged-HupR protein, a transcriptional regulator from the photosynthetic bacterium Rhodobacter capsulatus, were obtained by specific interaction with a Ni(2+) chelated lipid monolayer. HupR is a response regulator of the NtrC subfamily; it activates the transcription of the structural genes hupSLC, of [NiFe]hydrogenase. A projection map of the full-length protein at 9 A resolution was obtained by electron cryo-microscopy and image analysis of frozen-hydrated two-dimensional crystals. The crystals have a p6 plane group with unit cell dimensions of a=b=111.6(+/-1.0) A, gamma=120.4(+/-0.5) degrees. The structure of the N-terminal domain of NtrC, the family to which HupR belongs, had been determined previously by NMR. The atomic coordinates of the N-terminal domain of NtrC, were compared to the structure obtained by cryo-electron microscope techniques of the whole HupR. These results provide the first structure at medium resolution of a whole transcription factor, HupR from the NtrC family. PMID- 10677288 TI - Crystal structure of amylomaltase from thermus aquaticus, a glycosyltransferase catalysing the production of large cyclic glucans. AB - Amylomaltase is involved in the metabolism of starch, one of the most important polysaccharides in nature. A unique feature of amylomaltase is its ability to catalyze the formation of cyclic amylose. In contrast to the well studied cyclodextrin glucanotransferases (CGTases), which synthesize cycloamylose with a ring size (degree of polymerization or DP) of 6-8, the amylomaltase from Thermus aquaticus produces cycloamyloses with a DP of 22 and higher. The crystal structure of amylomaltase from Thermus aquaticus was determined to 2.0 A resolution. It is a member of the alpha-amylase superfamily of enzymes, whose core structure consists of a (beta, alpha)(8) barrel. In amylomaltase, the 8-fold symmetry of this barrel is disrupted by several insertions between the barrel strands. The largest insertions are between the third and fifth barrel strands, where two insertions form subdomain B1, as well as between the second and third barrel strands, forming the alpha-helical subdomain B2. Whereas part of subdomain B1 is also present in other enzyme structures of the alpha-amylase superfamily, subdomain B2 is unique to amylomaltase. Remarkably, the C-terminal domain C, which is present in all related enzymes of the alpha-amylase family, is missing in amylomaltase. Amylomaltase shows a similar arrangement of the catalytic side chains (two Asp residues and one Glu residue) as in previously characterized members of the alpha-amylase superfamily, indicating similar mechanisms of the glycosyl transfer reaction. In amylomaltase, a conserved loop of around eight amino acid residues is partially shielding the active center. This loop, which is well conserved among other amylomaltases, may sterically hinder the formation of small cyclic products. PMID- 10677289 TI - Binding of equine infectious anemia virus matrix protein to membrane bilayers involves multiple interactions. AB - Human immunodeficiency virus (HIV) and equine infectious anemia virus (EIAV) are closely related lentiviruses that infect immune cells, but their pathogenesis differ. Localization to the cytosolic leaflet of the plasma membrane is critical for replication of both viruses. This localization is accomplished through the matrix (MA) domain of the Gag precursor protein. In HIV-1, association of MA to anionic membranes appears to be primarily driven by a linear cluster of basic residues in the MA domain and an N-myristoylation signal. Interestingly, the MA protein of EIAV does not contain either of these signals. To understand which factors could promote EIAV assembly we characterized the membrane binding properties of its MA protein using fluorescence and biochemical methods. We find that EIAV MA exists as a multimer in solution whose protein-protein interactions are destabilized by membrane binding. EIAV MA binds strongly to electrically neutral membranes as well as to negatively charged membranes. Fluorescence quenching and chemical modification techniques, as well as trypsin proteolysis, indicate a different exposure of the EIAV MA Trp residues when bound to the two types of membranes, and EIAV MA proteolysis by trypsin differs when bound to the two types of membranes. Based on these data and the known structures of closely related matrix proteins, we constructed a structural model. This model predicts that EIAV MA binds to negatively charged membranes, but EIAV MA has an additional membrane binding region rich in residues that partition favorably into the membrane headgroup region. This secondary site may play a role in early events of viral infection. PMID- 10677290 TI - Photocrosslinking of benzophenone-labeled single cysteine troponin I mutants to other thin filament proteins. AB - The interaction sites of rabbit skeletal troponin I (TnI) with troponin C (TnC), troponin T (TnT), tropomyosin (Tm) and actin were mapped systematically using nine single cysteine residue TnI mutants with mutation sites at positions 6, 48, 64, 89, 104, 121, 133, 155 or 179 (TnI6, TnI48 etc.). Each mutant was labeled with the heterobifunctional photocrosslinker 4-maleimidobenzophenone (BP-Mal), and incorporated into the TnI.TnC binary complex, the TnI.TnC.TnT ternary troponin (Tn) complex, and the Tn.Tm.F-actin synthetic thin filament. Photocrosslinking reactions carried out in the presence and absence of Ca(2+) yielded the following results: (1) BP-TnI6 photocrosslinked primarily to TnC with a small degree of Ca(2+)-dependence in all the complex forms. (2) BP-TnI48, TnI64 and TnI89 photocrosslinked to TnT with no Ca(2+)-dependence. Photocrosslinking to TnC was reduced in the ternary versus the binary complex. BP-TnI89 also photocrosslinked to actin with higher yields in the absence of Ca(2+) than in its presence. (3) BP-TnI104 and TnI133 photocrosslinked to actin with much higher yields in the absence than in the presence of Ca(2+). (4) BP-TnI121 photocrosslinked to TnC with a small degree of Ca(2+)-dependence, and did not photocrosslink to actin. (5) BP-TnI155 and TnI179 photocrosslinked to TnC, TnT and actin, but all with low yields. All the labeled mutants photocrosslinked to TnC with varying degrees of Ca(2+)-dependence, and none to Tm. These results, along with those published allowed us to construct a structural and functional model of TnI in the Tn complex: in the presence of Ca(2+), residues 1-33 of TnI interact with the C-terminal domain hydrophobic cleft of TnC, approximately 48-89 with TnT, approximately 90-113 with TnC's central helix, approximately 114-125 with TnC's N-terminal domain hydrophobic cleft, and approximately 130-150 with TnC's A-helix. In the absence of Ca(2+), residues approximately 114-125 move out of TnC's N-terminal domain hydrophobic cleft and trigger the movements of residues approximately 89-113 and approximately 130-150 away from TnC and towards actin. PMID- 10677291 TI - The GxxxG motif: a framework for transmembrane helix-helix association. AB - In order to identify strong transmembrane helix packing motifs, we have selected transmembrane domains exhibiting high-affinity homo-oligomerization from a randomized sequence library based on the right-handed dimerization motif of glycophorin A. Sequences were isolated using the TOXCAT system, which measures transmembrane helix-helix association in the Escherichia coli inner membrane. Strong selection was applied to a large range of sequences ( approximately 10(7) possibilities) and resulted in the identification of sequence patterns that mediate high-affinity helix-helix association. The most frequent motif isolated, GxxxG, occurs in over 80% of the isolates. Additional correlations suggest that flanking residues act in concert with the GxxxG motif, and that size complementarity is maintained at the interface, consistent with the idea that the identified sequence patterns represent packing motifs. The convergent identification of similar sequence patterns from an analysis of the transmembrane domains in the SwissProt sequence database suggests that these packing motifs are frequently utilized in naturally occurring helical membrane proteins. PMID- 10677292 TI - Statistical analysis of amino acid patterns in transmembrane helices: the GxxxG motif occurs frequently and in association with beta-branched residues at neighboring positions. AB - To find motifs that mediate helix-helix interactions in membrane proteins, we have analyzed frequently occurring combinations of residues in a database of transmembrane domains. Our analysis was performed with a novel formalism, which we call TMSTAT, for exactly calculating the expectancies of all pairs and triplets of residues in individual sequences, taking into account differential sequence composition and the substantial effect of finite length in short segments. We found that the number of significantly over and under-represented pairs and triplets was much greater than the random expectation. Isoleucine, glycine and valine were the most common residues in these extreme cases. The main theme observed is patterns of small residues (Gly, Ala and Ser) at i and i+4 found in association with large aliphatic residues (Ile, Val and Leu) at neighboring positions (i.e. i+/-1 and i+/-2). The most over-represented pair is formed by two glycine residues at i and i+4 (GxxxG, 31.6 % above expectation, p<1x10(-33)) and it is strongly associated with the neighboring beta-branched residues Ile and Val. In fact, the GxxxG pair has been described as part of the strong interaction motif in the glycophorin A transmembrane dimer, in which the pair is associated with two Val residues (GVxxGV). GxxxG is also the major motif identified using TOXCAT, an in vivo selection system for transmembrane oligomerization motifs. In conjunction with these experimental observations, our results highlight the importance of the GxxxG+beta-branched motif in transmembrane helix-helix interactions. In addition, the special role for the beta-branched residues Ile and Val suggested here is consistent with the hypothesis that residues with constrained rotameric freedom in helical conformation might reduce the entropic cost of folding in transmembrane proteins. Additional material is available at http://engelman.csb.yale. edu/tmstat and http://bioinfo.mbb.yale. edu/tmstat. PMID- 10677293 TI - Design of nuclease resistant protein kinase calpha DNA enzymes with potential therapeutic application. AB - For the therapeutic application of catalytic nucleic acids it is desirable to have small, stable and inexpensive compounds that are active at physiological Mg(2+) concentrations. We have explored the possibility of using the versatile 10 23 DNA catalytic core to suppress the expression of the protein kinase Calpha (PKCalpha) isoform in malignant cells. By introducing either a 3'-3'-inverted thymidine nucleotide or site-specific phosphorothioate modification into a PKCalpha DNA enzyme, we have designed stable catalysts that retained a significant in vitro cleavage activity. In particular, a DNA enzyme containing phosphorothioate analogues in the antisense arms and in the pyrimidine residues of the catalytic core was found to be remarkably stable in 50 % human serum (t(1/2)>90 hours) and inhibited in vitro cell growth by up to 90 % at nanomolar concentrations. The inhibition of PKCalpha gene expression is sequence-specific, as a DNA enzyme with reversed antisense arms was found to be ineffective. Epifluorescence microscopic analysis of cells transfected with a 5' fluorescein isothiocyanate-conjugated DNA enzyme showed that the DNA enzyme molecules are mainly localised in the nuclei. Most of the DNA enzyme-treated cells were killed by apoptosis. The ability of the described PKCalpha DNA enzymes to trigger apoptosis (apoptozymes) in malignant cells illustrates their therapeutic potential. Furthermore, such agents can be a valuable tool for probing gene function. PMID- 10677294 TI - Methionine adenosyltransferase I/III deficiency: novel mutations and clinical variations. AB - Methionine adenosyltransferase (MAT) I/III deficiency, caused by mutations in the MAT1A gene, is characterized by persistent hypermethioninemia without elevated homocysteine or tyrosine. Clinical manifestations are variable and poorly understood, although a number of individuals with homozygous null mutations in MAT1A have neurological problems, including brain demyelination. We analyzed MAT1A in seven hypermethioninemic individuals, to provide insight into the relationship between genotype and phenotype. We identified six novel mutations and demonstrated that mutations resulting in high plasma methionines may signal clinical difficulties. Two patients-a compound heterozygote for truncating and severely inactivating missense mutations and a homozygote for an aberrant splicing MAT1A mutation-have plasma methionine in the 1,226-1,870 microM range (normal 5-35 microM) and manifest abnormalities of the brain gray matter or signs of brain demyelination. Another compound heterozygote for truncating and inactivating missense mutations has 770-1,240 microM plasma methionine and mild cognitive impairment. Four individuals carrying either two inactivating missense mutations or the single-allelic R264H mutation have 105-467 microM plasma methionine and are clinically unaffected. Our data underscore the necessity of further studies to firmly establish the relationship between genotypes in MAT I/III deficiency and clinical phenotypes, to elucidate the molecular bases of variability in manifestations of MAT1A mutations. PMID- 10677295 TI - Exon skipping in IVD RNA processing in isovaleric acidemia caused by point mutations in the coding region of the IVD gene. AB - Isovaleric acidemia (IVA) is a recessive disorder caused by a deficiency of isovaleryl-CoA dehydrogenase (IVD). We have reported elsewhere nine point mutations in the IVD gene in fibroblasts of patients with IVA, which lead to abnormalities in IVD protein processing and activity. In this report, we describe eight IVD gene mutations identified in seven IVA patients that result in abnormal splicing of IVD RNA. Four mutations in the coding region lead to aberrantly spliced mRNA species in patient fibroblasts. Three of these are amino acid altering point mutations, whereas one is a single-base insertion that leads to a shift in the reading frame of the mRNA. Two of the coding mutations strengthen pre-existing cryptic splice acceptors adjacent to the natural splice junctions and apparently interfere with exon recognition, resulting in exon skipping. This mechanism for missplicing has not been reported elsewhere. Four other mutations alter either the conserved gt or ag dinucleotide splice sites in the IVD gene. Exon skipping and cryptic splicing were confirmed by transfection of these mutations into a Cos-7 cell line model splicing system. Several of the mutations were predicted by individual information analysis to inactivate or significantly weaken adjacent donor or acceptor sites. The high frequency of splicing mutations identified in these patients is unusual, as is the finding of missplicing associated with missense mutations in exons. These results may lead to a better understanding of the phenotypic complexity of IVA, as well as provide insight into those factors important in defining intron/exon boundaries in vivo. PMID- 10677296 TI - Autosomal recessive disorder otospondylomegaepiphyseal dysplasia is associated with loss-of-function mutations in the COL11A2 gene. AB - Otospondylomegaepiphyseal dysplasia (OSMED) is an autosomal recessive skeletal dysplasia accompanied by severe hearing loss. The phenotype overlaps that of the autosomal dominant disorders-Stickler and Marshall syndromes-but can be distinguished by disproportionately short limbs, severe hearing loss, and lack of ocular involvement. In one family with OSMED, a homozygous Gly-->Arg substitution has been described in COL11A2, which codes for the alpha2 chain of type XI collagen. We report seven further families with OSMED. All affected individuals had a remarkably similar phenotype: profound sensorineural hearing loss, skeletal dysplasia with limb shortening and large epiphyses, cleft palate, an extremely flat face, hypoplasia of the mandible, a short nose with anteverted nares, and a flat nasal bridge. We screened affected individuals for mutations in COL11A2 and found different mutations in each family. Individuals from four families, including three with consanguineous parents, were homozygous for mutations. Individuals from three other families, in whom parents were nonconsanguineous, were compound heterozygous. Of the 10 identified mutations, 9 are predicted to cause premature termination of translation, and 1 is predicted to cause an in frame deletion. We conclude that the OSMED phenotype is highly homogenous and results from homozygosity or compound heterozygosity for COL11A2 mutations, most of which are predicted to cause complete absence of alpha2(XI) chains. PMID- 10677297 TI - Mutations in the AIRE gene: effects on subcellular location and transactivation function of the autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy protein. AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a monogenic autosomal disease with recessive inheritance. It is characterized by multiple autoimmune endocrinopathies, chronic mucocutaneous candidiasis, and ectodermal dystrophies. The defective gene responsible for this disease was recently isolated, and several different mutations in the novel gene, AIRE, have been identified, by us and by others, in patients with APECED. We have shown that the APECED protein is mainly localized, both in vitro and in vivo, to the cell nucleus, where it forms distinct speckles. This accords with the predicted structural features of the protein, which suggest involvement of AIRE in the regulation of gene transcription. Here, we report the results of mutational analyses of a series of 112 patients with APECED who were from various ethnic backgrounds. A total of 16 different mutations, covering 91% of disease alleles, were observed; of these, 8 were novel. The mutations are spread throughout the coding region of AIRE, yet four evident mutational hotspots were observed. In vitro expression of four different naturally occurring nonsense and missense mutations revealed a dramatically altered subcellular location of the protein in cultured cells. Interestingly, the wild-type APECED protein tethered to the Gal4 DNA-binding domain acted as a strong transcriptional activator of reporter genes in mammalian cells, whereas most of the analyzed mutant polypeptides had lost this capacity. PMID- 10677298 TI - Toward a survey of somatic mutation of the NF1 gene in benign neurofibromas of patients with neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1), a common autosomal dominant disorder caused by mutations of the NF1 gene, is characterized by multiple neurofibromas, pigmentation anomalies, and a variety of other possible complications, including an increased risk of malignant neoplasias. Tumorigenesis in NF1 is believed to follow the two-hit hypothesis postulated for tumor-suppressor genes. Loss of heterozygosity (LOH) has been shown to occur in NF1-associated malignancies and in benign neurofibromas, but only few of the latter yielded a positive result. Here we describe a systematic approach of searching for somatic inactivation of the NF1 gene in neurofibromas. In the course of these studies, two new intragenic polymorphisms of the NF1 gene, a tetranucleotide repeat and a 21-bp duplication, could be identified. Three tumor-specific point mutations and two LOH events were detected among seven neurofibromas from four different NF1 patients. Our results suggest that small subtle mutations occur with similar frequency to that of LOH in benign neurofibromas and that somatic inactivation of the NF1 gene is a general event in these tumors. The spectrum of somatic mutations occurring in various tumors from individual NF1 patients may contribute to the understanding of variable expressivity of the NF1 phenotype. PMID- 10677299 TI - Mutational spectrum in the Delta7-sterol reductase gene and genotype-phenotype correlation in 84 patients with Smith-Lemli-Opitz syndrome. AB - Smith-Lemli-Opitz syndrome (SLOS), an autosomal recessive malformation syndrome, ranges in clinical severity from mild dysmorphism and moderate mental retardation to severe congenital malformation and intrauterine lethality. Mutations in the gene for Delta7-sterol reductase (DHCR7), which catalyzes the final step in cholesterol biosynthesis in the endoplasmic reticulum (ER), cause SLOS. We have determined, in 84 patients with clinically and biochemically characterized SLOS (detection rate 96%), the mutational spectrum in the DHCR7 gene. Forty different SLOS mutations, some frequent, were identified. On the basis of mutation type and expression studies in the HEK293-derived cell line tsA-201, we grouped mutations into four classes: nonsense and splice-site mutations resulting in putative null alleles, missense mutations in the transmembrane domains (TM), mutations in the 4th cytoplasmic loop (4L), and mutations in the C-terminal ER domain (CT). All but one of the tested missense mutations reduced protein stability. Concentrations of the cholesterol precursor 7-dehydrocholesterol and clinical severity scores correlated with mutation classes. The mildest clinical phenotypes were associated with TM and CT mutations, and the most severe types were associated with 0 and 4L mutations. Most homozygotes for null alleles had severe SLOS; one patient had a moderate phenotype. Homozygosity for 0 mutations in DHCR7 appears compatible with life, suggesting that cholesterol may be synthesized in the absence of this enzyme or that exogenous sources of cholesterol can be used. PMID- 10677300 TI - Imprinting effect in premature ovarian failure confined to paternally inherited fragile X premutations. AB - Fragile X premutations are considered to be a risk factor for premature ovarian failure (POF), which is usually defined as menopause at age <40 years. Since premutations may be inherited from either the mother or the father, we evaluated the influence of the inheritance pattern on the duration of reproductive life in female carriers. The occurrence of POF and age at menopause in women with a paternally inherited fragile X premutation (PIP) were compared to those in women with a maternally inherited fragile X premutation (MIP). We identified 148 women in whom the parental origin of the premutation could be determined. In 109 of these women we were able to establish whether POF had occurred: 82 women had a PIP, and 27 had a MIP. Twenty-three of the women (28%) with a PIP had POF, versus only 1 (3.7%) with a MIP (two -tailed Fisher's exact test; P=. 007). Kaplan-Meier analysis of all 148 premutations showed that the age at menopause was significantly lower in the women with a PIP than in the woman with a MIP (Breslow test in Kaplan-Meier analysis; P=.003). Our data strongly suggest that, when POF occurs in fragile X premutation carriers, a considerable proportion of the premutations are inherited paternally (parent-of-origin effect). We hypothesize that this may be owing to a paternal genomic imprinting effect. PMID- 10677301 TI - Detection of chromosomal aberrations by a whole-genome microsatellite screen. AB - Chromosomal aberrations are a common cause of multiple anomaly syndromes that include developmental and growth retardation. Current microscopic techniques are useful for the detection of such aberrations but have a limit of resolution that is above the threshold for phenotypic effect. We hypothesized that a genomewide microsatellite screen could detect chromosomal aberrations that were not detected by standard cytogenetic techniques in a portion of these individuals. To test this hypothesis, we performed a genomewide microsatellite screen of patients, by use of a currently available genetic-marker panel that was originally designed for meiotic mapping of Mendelian traits. We genotyped approximately 400 markers on 17 pairs of parents and their children who had normal karyotypes. By using this approach, we detected and confirmed two cases of segmental aneusomy among 11 children with multiple congenital anomalies. These data demonstrate that a genomewide microsatellite scan can be used to detect chromosomal aberrations that are not detected by microscopic techniques. PMID- 10677302 TI - Assignment of a form of congenital muscular dystrophy with secondary merosin deficiency to chromosome 1q42. AB - We have previously reported an autosomal recessive form of congenital muscular dystrophy, characterized by proximal girdle weakness, generalized muscle hypertrophy, rigidity of the spine, and contractures of the tendo Achilles, in a consanguineous family from the United Arab Emirates. Early respiratory failure resulting from severe diaphragmatic involvement was present. Intellect and the results of brain imaging were normal. Serum creatine kinase levels were grossly elevated, and muscle-biopsy samples showed dystrophic changes. The expression of the laminin-alpha2 chain of merosin was reduced on several fibers, but linkage analysis excluded the LAMA2 locus on chromosome 6q22-23. Here, we report the results of genomewide linkage analysis of this family, by use of homozygosity mapping. In all four affected children, an identical homozygous region was identified on chromosome 1q42, spanning 6-15 cM between flanking markers D1S2860 and D1S2800. We have identified a second German family with two affected children having similar clinical and histopathological features; they are consistent with linkage to the same locus. The cumulative LOD score was 3.57 (straight theta=.00) at marker D1S213. This represents a novel locus for congenital muscular dystrophy. We suggest calling this disorder "CMD1B." The expression of three functional candidate genes in the CMD1B critical region was investigated, and no detectable changes in their level of expression were observed. The secondary reduction in laminin-alpha2 chain in these families suggests that the primary genetic defect resides in a gene coding for a protein involved in basal lamina assembly. PMID- 10677303 TI - Familial syndromic esophageal atresia maps to 2p23-p24. AB - Esophageal atresia (EA) is a common life-threatening congenital anomaly that occurs in 1/3,000 newborns. Little is known of the genetic factors that underlie EA. Oculodigitoesophageoduodenal (ODED) syndrome (also known as "Feingold syndrome") is a rare autosomal dominant disorder with digital abnormalities, microcephaly, short palpebral fissures, mild learning disability, and esophageal/duodenal atresia. We studied four pedigrees, including a three generation Dutch family with 11 affected members. Linkage analysis was initially aimed at chromosomal regions harboring candidate genes for this disorder. Twelve different genomic regions covering 15 candidate genes (approximately 15% of the genome) were excluded from involvement in the ODED syndrome. A subsequent nondirective mapping approach revealed evidence for linkage between the syndrome and marker D2S390 (maximum LOD score 4.51 at recombination fraction 0). A submicroscopic deletion in a fourth family with ODED provided independent confirmation of this genetic localization and narrowed the critical region to 7.3 cM in the 2p23-p24 region. These results show that haploinsufficiency for a gene or genes in 2p23-p24 is associated with syndromic EA. PMID- 10677304 TI - Localization of the gene for a novel autosomal recessive neurodegenerative Huntington-like disorder to 4p15.3. AB - A consanguineous family affected by an autosomal recessive, progressive neurodegenerative Huntington-like disorder, was tested to rule out juvenile-onset Huntington disease (JHD). The disease manifests at approximately 3-4 years and is characterized by both pyramidal and extrapyramidal abnormalities, including chorea, dystonia, ataxia, gait instability, spasticity, seizures, mutism, and intellectual impairment. Brain magnetic resonance imaging (MRI) findings include progressive frontal cortical atrophy and bilateral caudate atrophy. Huntington CAG trinucleotide-repeat analyses ruled out JHD, since all affected individuals had repeat numbers within the normal range. The presence of only four recombinant events (straight theta=.2) between the disease and the Huntington locus in 20 informative meioses suggested that the disease localized to chromosome 4. Linkage was initially achieved with marker D4S2366 at 4p15.3 (LOD 3.03). High-density mapping at the linked locus resulted in homozygosity for markers D4S431 and D4S394, which span a 3-cM region. A maximum LOD score of 4.71 in the homozygous interval was obtained. Heterozygosity at the distal D4S2366 and proximal D4S2983 markers defines the maximum localization interval (7 cM). Multiple brain-related expressed sequence tags (ESTs) with no known disease association exist in the linkage interval. Among the three known genes residing in the linked interval (ACOX3, DRD5, QDPR), the most likely candidate, DRD5, encoding the dopamine receptor D5, was excluded, since all five affected family members were heterozygous for an intragenic dinucleotide repeat. The inheritance pattern and unique localization to 4p15.3 are consistent with the identification of a novel, autosomal recessive, neurodegenerative Huntington-like disorder. PMID- 10677305 TI - A new locus for autosomal recessive hypercholesterolemia maps to human chromosome 15q25-q26. AB - High serum cholesterol is an established risk factor for cardiovascular disease and is the prime target for therapeutic intervention in large groups of patients. The development of modern treatments for this major risk factor was propelled by the early realization that forms of severe hypercholesterolemia could be caused by dominantly inherited defects in the LDL receptor or in the APOB gene. Further understanding of the mechanisms contributing to early atherosclerosis will allow for new targets for therapy. We therefore identified and investigated the genetics of families from Sardinia that have recessive inheritance of precocious hypercholesterolemia. We used five families in an analysis of linkage of the autosomal recessive hypercholesterolemia locus, termed "ARH1," to chromosome 15q25-q26. A genomewide search mapped the disease-causing gene with a LOD score of 3.3 and excluded major contributions to the phenotype of other genes. A candidate gene present in the mapped chromosome region-the ligand-activated liver transcription-factor gene ARP1 (apolipoprotein regulatory-protein gene)-has been excluded after DNA sequencing. The close-bred nature of the Sardinian population offers unique opportunities for isolation of this hypercholesterolemia-causing gene. PMID- 10677306 TI - X-Linked syndrome of polyendocrinopathy, immune dysfunction, and diarrhea maps to Xp11.23-Xq13.3. AB - We describe genetic analysis of a large pedigree with an X-linked syndrome of polyendocrinopathy, immune dysfunction, and diarrhea (XPID), which frequently results in death during infancy or childhood. Linkage analysis mapped the XPID gene to a 17-cM interval defined by markers DXS8083 and DXS8107 on the X chromosome, at Xp11. 23-Xq13.3. The maximum LOD score was 3.99 (recombination fraction0) at DXS1235. Because this interval also harbors the gene for Wiskott Aldrich syndrome (WAS), we investigated mutations in the WASP gene, as the molecular basis of XPID. Northern blot analysis detected the same relative amount and the same-sized WASP message in patients with XPID and in a control. Analysis of the WASP coding sequence, an alternate promoter, and an untranslated upstream first exon was carried out, and no mutations were found in patients with XPID. A C-->T transition within the alternate translation start site cosegregated with the XPID phenotype in this family; however, the same transition site was detected in a normal control male. We conclude that XPID maps to Xp11.23-Xq13.3 and that mutations of WASP are not associated with XPID. PMID- 10677307 TI - A unique form of mental retardation with a distinctive phenotype maps to Xq26 q27. AB - We report a novel X-linked mental retardation (XLMR) syndrome, with characteristic facial dysmorphic features, segregating in a large North Carolina family. Only males are affected, over four generations. Clinical findings in the seven living affected males include a moderate degree of mental retardation (MR), coarse facies, puffy eyelids, narrow palpebral fissures, prominent supraorbital ridges, a bulbous nose, a prominent lower lip, large ears, obesity, and large testicles. Cephalometric measurements suggest that the affected males have a distinctive craniofacial skeletal structure, when compared with normative measures. Obligate-carrier females are unaffected with MR, but the results of cephalometric skeletal analysis suggest craniofacial dysmorphisms intermediate between affected males and normative control individuals. Unaffected male relatives show no clinical or cephalometric resemblance to affected males. The blood-lymphocyte karyotype and the results of DNA analysis for fragile-X syndrome and of other routine investigations are normal. Linkage analysis for polymorphic DNA markers spanning the X chromosome established linkage to Xq26-q27. Maximum LOD scores were obtained at marker DXS1047 (maximum LOD score = 3.1 at recombination fraction 0). By use of haplotype analysis, we have localized the gene for this condition to an 18-cM genetic interval flanked by ATA59C05 and GATA31E08. On the basis of both the clinical phenotype and the mapping data, we were able to exclude other reported XLMR conditions. Therefore, we believe that a unique recessive XLMR syndrome with a distinctive and recognizable phenotype is represented in this family. PMID- 10677308 TI - Survey of the fragile X syndrome CGG repeat and the short-tandem-repeat and single-nucleotide-polymorphism haplotypes in an African American population. AB - Previous studies have shown that specific short-tandem-repeat (STR) and single nucleotide-polymorphism (SNP)-based haplotypes within and among unaffected and fragile X white populations are found to be associated with specific CGG-repeat patterns. It has been hypothesized that these associations result from different mutational mechanisms, possibly influenced by the CGG structure and/or cis-acting factors. Alternatively, haplotype associations may result from the long mutational history of increasing instability. To understand the basis of the mutational process, we examined the CGG-repeat size, three flanking STR markers (DXS548-FRAXAC1-FRAXAC2), and one SNP (ATL1) spanning 150 kb around the CGG repeat in unaffected (n=637) and fragile X (n=63) African American populations and compared them with unaffected (n=721) and fragile X (n=102) white populations. Several important differences were found between the two ethnic groups. First, in contrast to that seen in the white population, no associations were observed among the African American intermediate or "predisposed" alleles (41-60 repeats). Second, two previously undescribed haplotypes accounted for the majority of the African American fragile X population. Third, a putative "protective" haplotype was not found among African Americans, whereas it was found among whites. Fourth, in contrast to that seen in whites, the SNP ATL1 was in linkage equilibrium among African Americans, and it did not add new information to the STR haplotypes. These data indicate that the STR- and SNP based haplotype associations identified in whites probably reflect the mutational history of the expansion, rather than a mutational mechanism or pathway. PMID- 10677309 TI - ATM-heterozygous germline mutations contribute to breast cancer-susceptibility. AB - Approximately 0.5%-1% of the general population has been estimated to be heterozygous for a germline mutation in the ATM gene. Mutations in the ATM gene are responsible for the autosomal recessive disorder ataxia-telangiectasia (A-T) (MIM 208900). The finding that ATM-heterozygotes have an increased relative risk for breast cancer was supported by some studies but not confirmed by others. In view of this discrepancy, we examined the frequency of ATM germline mutations in a selected group of Dutch patients with breast cancer. We have analyzed ATM germline mutations in normal blood lymphocytes, using the protein-truncation test followed by genomic-sequence analysis. A high percentage of ATM germline mutations was demonstrated among patients with sporadic breast cancer. The 82 patients included in this study had developed breast cancer at age <45 and had survived >/=5 years (mean 15 years), and in 33 (40%) of the patients a contralateral breast tumor had been diagnosed. Among these patients we identified seven (8.5%) ATM germline mutations, of which five are distinct. One splice-site mutation (IVS10-6T-->G) was detected three times in our series. Four heterozygous carriers were patients with bilateral breast cancer. Our results indicate that the mutations identified in this study are "A-T disease-causing" mutations that might be associated with an increased risk of breast cancer in heterozygotes. We conclude that ATM heterozygotes have an approximately ninefold-increased risk of developing a type of breast cancer characterized by frequent bilateral occurrence, early age at onset, and long-term survival. The specific characteristics of our population of patients may explain why such a high frequency was not found in other series. PMID- 10677310 TI - Fine localization of a major disease-susceptibility locus for diffuse panbronchiolitis. AB - Diffuse panbronchiolitis affecting East Asians is strongly associated with the class I human leukocyte antigen (HLA) alleles. Recent observations suggest that a major disease-susceptibility gene may be located between the HLA-B and HLA-A loci in the class I region of the major histocompatibility complex on chromosome 6. To test this possibility, we analyzed 14 polymorphic markers in 92 Japanese patients and 93 healthy controls. Of these, seven marker alleles, including HLA-B54 and HLA-A11, were significantly associated with the disease. Maximum-likelihood haplotype analysis and subsequent direct determination of individual haplotypes identified a group of disease-associated haplotypes, one of which contained all seven disease-associated marker alleles. Another haplotype, containing HLA B*5504, was also associated with the disease. All these haplotypes seem to have diverged from a common ancestral haplotype in East Asians and share a specific segment containing three consecutive markers between the S and TFIIH loci in the class I region. Furthermore, one of the markers within the candidate region showed the highest delta value, indicating the strongest association. Of 20 Korean patients with diffuse panbronchiolitis, 17 also shared the combination of the disease-associated marker alleles within the candidate region. These results indicate that an HLA-associated major susceptibility gene for diffuse panbronchiolitis is probably located within the 200 kb in the class I region 300 kb telomeric of the HLA-B locus on the chromosome 6p21.3. PMID- 10677311 TI - Reproducibility and complications in gene searches: linkage on chromosome 6, heterogeneity, association, and maternal inheritance in juvenile myoclonic epilepsy. AB - Evidence for genetic influences in epilepsy is strong, but reports identifying specific chromosomal origins of those influences conflict. One early study reported that human leukocyte antigen (HLA) markers were genetically linked to juvenile myoclonic epilepsy (JME); this was confirmed in a later study. Other reports did not find linkage to HLA markers. One found evidence of linkage to markers on chromosome 15, another to markers on chromosome 6, centromeric to HLA. We identified families through a patient with JME and genotyped markers throughout chromosome 6. Linkage analysis assuming equal male-female recombination probabilities showed evidence for linkage (LOD score 2.5), but at a high recombination fraction (theta), suggesting heterogeneity. When linkage analysis was redone to allow independent male-female thetas, the LOD score was significantly higher (4.2) at a male-female theta of.5,.01. Although the overall pattern of LOD scores with respect to male-female theta could not be explained solely by heterogeneity, the presence of heterogeneity and predominantly maternal inheritance of JME might explain it. By analyzing loci between HLA-DP and HLA-DR and stratifying the families on the basis of evidence for or against linkage, we were able to show evidence of heterogeneity within JME and to propose a marker associated with the linked form. These data also suggest that JME may be predominantly maternally inherited and that the HLA-linked form is more likely to occur in families of European origin. PMID- 10677312 TI - Variation in the interleukin 4-receptor alpha gene confers susceptibility to asthma and atopy in ethnically diverse populations. AB - After a genomewide screen in the Hutterites was completed, the IL4RA gene was examined as the 16p-linked susceptibility locus for asthma and atopy. Seven known variants and one novel variant, representing all nonsynonymous substitutions in the mature protein, were examined in the Hutterites; on the basis of studies in the Hutterites, outbred white, black, and Hispanic families were genotyped for selected markers. All population samples showed evidence of association to atopy or to asthma (P values.039-.0044 for atopy and. 029-.0000061 for asthma), but the alleles or haplotypes showing the strongest evidence differed between the groups. Overall, these data suggest that the IL4RA gene is an atopy- and asthma susceptibility locus but that variation outside the coding region of the gene influences susceptibility. PMID- 10677313 TI - Individual estimates of European genetic admixture associated with lower body mass index, plasma glucose, and prevalence of type 2 diabetes in Pima Indians. AB - Individual genetic admixture estimates (IA) from European Americans (EAs) were computed in 7,996 members of the Gila River Indian Community (Arizona). Parental populations for the analysis were European Americans and full-heritage Pima Indians. A logistic regression was performed on 7,796 persons, to assess association of IA with type 2 diabetes. The odds ratio, comparing diabetes risk in full-heritage EAs with full-heritage Pima Indians, was 0.329 (95% confidence interval [CI] 0.225-0.482). Proportional-hazards analysis was performed on 5,482 persons who were nondiabetic at their first examination and 1,215 subjects who developed diabetes during the study. The hazard risk ratio for IA was 0.455 (95% CI 0.301-0.688). Nondiabetic persons had significantly more European IA. In nondiabetic Pimans, multivariate linear regressions of quantitative predictors of type 2 diabetes mellitus, including fasting plasma glucose, 2-h post-load plasma glucose, and body-mass index, showed significant inverse relations with IA when controlled for sex and age. These results illustrate the ongoing evolution of populations by the mechanism of gene flow and its effect on disease risk in the groups with admixture. When the two parental populations differ in disease prevalence, higher or lower risk is associated with admixture, depending on the origin of the admixed alleles and the relative magnitude of the disease prevalence in the parental populations. These data also illustrate the strong genetic components in type 2 diabetes and are consistent with one susceptibility locus common to obesity and diabetes. PMID- 10677314 TI - Linkage analyses at the chromosome 1 loci 1q24-25 (HPC1), 1q42.2-43 (PCAP), and 1p36 (CAPB) in families with hereditary prostate cancer. AB - Recent studies suggest that hereditary prostate cancer (PRCA) is a complex disease, involving multiple susceptibility genes and variable phenotypic expression. Through linkage analysis, potential prostate cancer susceptibility loci have been mapped to 3 regions on chromosome 1. To investigate the reported linkage to these regions, we conducted linkage studies on 144 PRCA families by using microsatellite markers in regions 1q24-25 (HPC1) and 1q42.2-43 (PCAP). We also examined the 1p36 (CAPB) region in 13 PRCA families with at least one case of brain cancer. No significant evidence of linkage to the HPC1 or PCAP region was found when the entire data set was analyzed. However, weak evidence for linkage to HPC1 was observed in the subset of families with male-to-male transmission (n=102; maximum multipoint nonparametric linkage [NPL] 1.99, P=.03). Weak evidence for linkage with heterogeneity within this subset was also observed (HLOD 1.21, P=.02), with approximately 20% of families linked. Although not statistically significant, suggestive evidence for linkage to PCAP was observed for the families (n=21) that met the three criteria of male-to-male transmission, average age of diagnosis <66 years, and >/=5 affected individuals (maximum multipoint NPL 1.45, P=.08). There was no evidence for linkage to CAPB in the brain cancer-prostate cancer subset. These results strengthen the argument that prostate cancer is a heterogeneous disease and that multiple genetic and environmental factors may be important for its etiology. PMID- 10677315 TI - Genome screening in human systemic lupus erythematosus: results from a second Minnesota cohort and combined analyses of 187 sib-pair families. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a loss of immunologic tolerance to a multitude of self-antigens. Epidemiological data suggest an important role for genes in the etiology of lupus, and previous genetic studies have implicated the HLA locus, complement genes, and low-affinity IgG (Fcgamma) receptors in SLE pathogenesis. In an effort to identify new susceptibility loci for SLE, we recently reported the results of a genomewide microsatellite marker screen in 105 SLE sib-pair families. By using nonparametric methods, evidence for linkage was found in four intervals: 6p11-21 (near the HLA), 16q13, 14q21-23, and 20p12.3 (LOD scores >/=2.0), and weaker evidence in another nine regions. We now report the results of a second complete genome screen in a new cohort of 82 SLE sib-pair families. In the cohort 2 screen, the four best intervals were 7p22 (LOD score 2.87), 7q21 (LOD score 2.40), 10p13 (LOD score 2.24), and 7q36 (LOD score 2.15). Eight additional intervals were identified with LOD scores in the range 1.00-1.67. A combined analysis of MN cohorts 1 and 2 (187 sib-pair families) showed that markers in 6p11-p21 (D6S426, LOD score 4.19) and 16q13 (D16S415, LOD score 3.85) met the criteria for significant linkage. Three intervals (2p15, 7q36, and 1q42) had LOD scores in the range 1.92-2.06, and another 13 intervals had LOD scores in the range of 1.00 1.78 in the combined sample. These data, together with other available gene mapping results in SLE, are beginning to allow a prioritization of genomic intervals for gene discovery efforts in human SLE. PMID- 10677316 TI - Evidence for heterogeneity in recombination in the human pseudoautosomal region: high resolution analysis by sperm typing and radiation-hybrid mapping. AB - Accurate genetic and physical maps for the human pseudoautosomal region were constructed by use of sperm typing and high-resolution radiation-hybrid mapping. PCR analysis of 1,912 sperm was done with a manual, single-sperm isolation method. Data on four donors show highly significant linkage heterogeneity among individuals. The most significant difference was observed in a marker interval located in the middle of the Xp/Yp pseudoautosomal region, where one donor showed a particularly high recombination fraction. Longitudinal models were fitted to the data to test whether linkage heterogeneity among donors was significant for multiple intervals across the region. The results indicated that increased recombination in particular individuals and regions is compensated for by reduced recombination in neighboring intervals. To investigate correspondence between physical and genetic distances within the region, we constructed a high resolution radiation-hybrid map containing 29 markers. The recombination fraction per unit of physical distance varies between regions ranging from 13- to 70-fold greater than the genome-average rate. PMID- 10677317 TI - Effects of stratification in the analysis of affected-sib-pair data: benefits and costs. AB - The benefits and costs of stratification of affected-sib-pair (ASP) data were examined in three situations: (1) when there is no difference in identity-by descent (IBD) allele sharing between stratified and unstratified ASP data sets; (2) when there is an increase in IBD allele sharing in one of the stratified groups; and (3) when the data are stratified on the basis of IBD allele-sharing status at one locus, and the stratified ASPs are then analyzed for linkage at a second locus. When there is no difference in IBD sharing between strata, a penalty is always paid for stratifying the data. The loss of power to detect linkage in the stratified ASP data sets is the result of multiple testing and the smaller sample size within individual strata. In the case in which etiologic heterogeneity (i.e., severity of phenotype, age at onset) represents genetic heterogeneity, the power to detect linkage can be increased by stratifying the ASP data. This benefit is obtained when there is sufficient IBD allele sharing and sample sizes. Once linkage has been established for a given locus, data can be stratified on the basis of IBD status at this locus and can be tested for linkage at a second locus. When the relative risk is in the vicinity of 1, the power to detect linkage at the second locus is always greater for the unstratified ASP data set. Even for values of the relative risk that diverge sufficiently from 1, with adequate sample sizes and IBD allele sharing, the benefits of stratifying ASP data are minimal. PMID- 10677318 TI - Removing the sampling restrictions from family-based tests of association for a quantitative-trait locus. AB - One strategy for localization of a quantitative-trait locus (QTL) is to test whether the distribution of a quantitative trait depends on the number of copies of a specific genetic-marker allele that an individual possesses. This approach tests for association between alleles at the marker and the QTL, and it assumes that association is a consequence of the marker being physically close to the QTL. However, problems can occur when data are not from a homogeneous population, since associations can arise irrespective of a genetic marker being in physical proximity to the QTL-that is, no information is gained regarding localization. Methods to address this problem have recently been proposed. These proposed methods use family data for indirect stratification of a population, thereby removing the effect of associations that are due to unknown population substructure. They are, however, restricted in terms of the number of children per family that can be used in the analysis. Here we introduce tests that can be used on family data with parent and child genotypes, with child genotypes only, or with a combination of these types of families, without size restrictions. Furthermore, equations that allow one to determine the sample size needed to achieve desired power are derived. By means of simulation, we demonstrate that the existing tests have an elevated false-positive rate when the size restrictions are not followed and that a good deal of information is lost as a result of adherence to the size restrictions. Finally, we introduce permutation procedures that are recommended for small samples but that can also be used for extensions of the tests to multiallelic markers and to the simultaneous use of more than one marker. PMID- 10677319 TI - The relationship between the sibling recurrence-risk ratio and genotype relative risk. AB - The recurrence-risk ratio of disease in siblings, lambdaS, is a standard parameter used in genetic analysis to estimate the statistical power for detection of a disease locus. However, the relationship between the underlying risk conferred by a disease-susceptibility allele and lambdaS has not been well described. The former is generally quantified as a genotype relative risk, gamma, and measures the ratio of disease risks between those with and those without the susceptibility genotype(s). We demonstrate that lambdaS varies significantly more with respect to gamma and the disease-allele frequency for two-locus multiplicative models than for other two-locus and for single-locus models. For the single- and two-locus dominant-inheritance models that we studied, when a disease-susceptibility allele had a frequency >/=.2, lambdaS had an upper limit of <10. In general, lambdaS values >10 are possible only under recessive inheritance, dominant inheritance with relatively rare (<5%) disease susceptibility alleles, or when two or more disease loci have alleles acting either epistatically or multiplicatively. We introduce the idea of a restricted sib recurrence-risk ratio (lambda*S) estimated by restriction of sibships to those ascertained through a proband who already has a putative high-risk allele. A lambda*S larger than the lambdaS value estimated from randomly selected probands can serve as an indirect way of testing whether the posited susceptibility allele increases disease risk. Our results demonstrate that a lambdaS of 2-3 may portend successful mapping for a variety of genetic models but that, for some two-locus models, a lambdaS as high as 10 does not guarantee underlying genes easily mapped by linkage. PMID- 10677320 TI - Family-based tests of association and linkage that use unaffected sibs, covariates, and interactions. AB - We extend the methodology for family-based tests of association and linkage to allow for both variation in the phenotypes of subjects and incorporation of covariates into general-score tests of association. We use standard association models for a phenotype and any number of predictors. We then construct a score statistic, using likelihoods for the distribution of phenotype, given genotype. The distribution of the score is computed as a function of offspring genotypes, conditional on parental genotypes and trait values for offspring and parents. This approach provides a natural extension of the transmission/disequilibrium test to any phenotype and to multiple genes or environmental factors and allows the study of gene-gene and gene-environment interaction. When the trait varies among subjects or when covariates are included in the association model, the score statistic depends on one or more nuisance parameters. We suggest two approaches for obtaining parameter estimates: (1) choosing the estimate that minimizes the variance of the test statistic and (2) maximizing the statistic over a nuisance parameter and using a corrected P value. We apply our methods to a sample of families with attention-deficit/hyperactivity disorder and provide examples of how covariates and gene-environment and gene-gene interactions can be incorporated. PMID- 10677321 TI - A coalescent approach to study linkage disequilibrium between single-nucleotide polymorphisms. AB - We present the results of extensive simulations that emulate the development and distribution of linkage disequilibrium (LD) between single-nucleotide polymorphisms (SNPs) and a gene locus that is phenotypically stratified into two classes (disease phenotype and wild-type phenotype). Our approach, based on coalescence theory, allows an explicit modeling of the demographic history of the population without conditioning on the age of the mutation, and serves as an efficient tool to carry out simulations. More specifically, we compare the influence that a constant population size or an exponentially growing population has on the amount of LD. These results indicate that attempts to locate single disease genes are most likely successful in small and constant populations. On the other hand, if we consider an exponentially growing population that started to expand from an initially constant population of reasonable size, then our simulations indicate a lower success rate. The power to detect association is enhanced if haplotypes constructed from several SNPs are used as markers. The versatility of the coalescence approach also allows the analysis of other relevant factors that influence the chances that a disease gene will be located. We show that several alleles leading to the same disease have no substantial influence on the amount of LD, as long as the differences between the disease causing alleles are confined to the same region of the gene locus and as long as each allele occurs in an appreciable frequency. Our simulations indicate that mapping of less-frequent diseases is more likely to be successful. Moreover, we show that successful attempts to map complex diseases depend crucially on the phenotype-genotype correlations of all alleles at the disease locus. An analysis of lipoprotein lipase data indicates that our simulations capture the major features of LD occurring in biological data. PMID- 10677322 TI - Estimation of variance components of quantitative traits in inbred populations. AB - Use of variance-component estimation for mapping of quantitative-trait loci in humans is a subject of great current interest. When only trait values, not genotypic information, are considered, variance-component estimation can also be used to estimate heritability of a quantitative trait. Inbred pedigrees present special challenges for variance-component estimation. First, there are more variance components to be estimated in the inbred case, even for a relatively simple model including additive, dominance, and environmental effects. Second, more identity coefficients need to be calculated from an inbred pedigree in order to perform the estimation, and these are computationally more difficult to obtain in the inbred than in the outbred case. As a result, inbreeding effects have generally been ignored in practice. We describe here the calculation of identity coefficients and estimation of variance components of quantitative traits in large inbred pedigrees, using the example of HDL in the Hutterites. We use a multivariate normal model for the genetic effects, extending the central-limit theorem of Lange to allow for both inbreeding and dominance under the assumptions of our variance-component model. We use simulated examples to give an indication of under what conditions one has the power to detect the additional variance components and to examine their impact on variance-component estimation. We discuss the implications for mapping and heritability estimation by use of variance components in inbred populations. PMID- 10677323 TI - New estimates of intergenerational time intervals for the calculation of age and origins of mutations. AB - Intergenerational time intervals are frequently used in human population-genetics studies concerned with the ages and origins of mutations. In most cases, mean intervals of 20 or 25 years are used, regardless of the demographic characteristics of the population under study. Although these characteristics may vary from prehistoric to historical times, we suggest that this value is probably too low, and that the ages of some mutations may have been underestimated. Analyses were performed by using the BALSAC Population Register (Quebec, Canada), from which several intergenerational comparisons can be made. Family reconstitutions were used to measure interval lengths and variations in descending lineages. Various parameters were considered, such as spouse age at marriage, parental age, and reproduction levels. Mother-child and father-child intervals were compared. Intergenerational male and female intervals were also analyzed in 100 extended ascending genealogies. Results showed that a mean value of 30 years is a better estimate of intergenerational intervals than 20 or 25 years. As marked differences between male and female interval length were observed, specific values are proposed for mtDNA, autosomal, X-chromosomal, and Y chromosomal loci. The applicability of these results for age estimates of mutations is discussed. PMID- 10677324 TI - Haplotype fine mapping by evolutionary trees. AB - To refine the location of a disease gene within the bounds provided by linkage analysis, many scientists use the pattern of linkage disequilibrium between the disease allele and alleles at nearby markers. We describe a method that seeks to refine location by analysis of "disease" and "normal" haplotypes, thereby using multivariate information about linkage disequilibrium. Under the assumption that the disease mutation occurs in a specific gap between adjacent markers, the method first combines parsimony and likelihood to build an evolutionary tree of disease haplotypes, with each node (haplotype) separated, by a single mutational or recombinational step, from its parent. If required, latent nodes (unobserved haplotypes) are incorporated to complete the tree. Once the tree is built, its likelihood is computed from probabilities of mutation and recombination. When each gap between adjacent markers is evaluated in this fashion and these results are combined with prior information, they yield a posterior probability distribution to guide the search for the disease mutation. We show, by evolutionary simulations, that an implementation of these methods, called "FineMap," yields substantial refinement and excellent coverage for the true location of the disease mutation. Moreover, by analysis of hereditary hemochromatosis haplotypes, we show that FineMap can be robust to genetic heterogeneity. PMID- 10677325 TI - Y chromosomes traveling south: the cohen modal haplotype and the origins of the Lemba--the "Black Jews of Southern Africa". AB - The Lemba are a traditionally endogamous group speaking a variety of Bantu languages who live in a number of locations in southern Africa. They claim descent from Jews who came to Africa from "Sena." "Sena" is variously identified by them as Sanaa in Yemen, Judea, Egypt, or Ethiopia. A previous study using Y chromosome markers suggested both a Bantu and a Semitic contribution to the Lemba gene pool, a suggestion that is not inconsistent with Lemba oral tradition. To provide a more detailed picture of the Lemba paternal genetic heritage, we analyzed 399 Y chromosomes for six microsatellites and six biallelic markers in six populations (Lemba, Bantu, Yemeni-Hadramaut, Yemeni-Sena, Sephardic Jews, and Ashkenazic Jews). The high resolution afforded by the markers shows that Lemba Y chromosomes are clearly divided into Semitic and Bantu clades. Interestingly, one of the Lemba clans carries, at a very high frequency, a particular Y-chromosome type termed the "Cohen modal haplotype," which is known to be characteristic of the paternally inherited Jewish priesthood and is thought, more generally, to be a potential signature haplotype of Judaic origin. The Bantu Y-chromosome samples are predominantly (>80%) YAP+ and include a modal haplotype at high frequency. Assuming a rapid expansion of the eastern Bantu, we used variation in microsatellite alleles in YAP+ sY81-G Bantu Y chromosomes to calculate a rough date, 3,000-5,000 years before the present, for the start of their expansion. PMID- 10677326 TI - The X chromosome frequently lags behind in female lymphocyte anaphase. AB - Pancentromeric FISH and X-chromosome painting were used to characterize anaphase aberrations in 2,048 cultured lymphocytes from a healthy 62-year-old woman. Of 163 aberrant anaphases, 66.9% contained either chromosomes or their fragments that lagged behind. Characterization of 200 laggards showed that 49% were autosomes, 33. 5% were autosomal fragments, and 17.5% were X chromosomes. The X chromosome represented one-fourth of all lagging chromosomes and was involved much more often than would be expected by chance (1/23). Labeling of the late replicating inactive X chromosome with 5-bromo-2'-deoxyuridine revealed that both X homologues contributed equally to the laggards. Among 200 micronuclei examined from interphase cells, the proportion of the X chromosome (31%) and autosomal fragments (50%) was higher than among anaphase laggards, whereas autosomes were involved less often (19%). These findings may reflect either selection or the fact that lagging autosomes, which were more proximal to the poles than were lagging X chromosomes, were more frequently included within the main nucleus. Our results suggest that the well-known high micronucleation and loss of the X chromosome in women's lymphocytes is the result of frequent distal lagging behind in anaphase and effective micronucleation of this chromosome. This lagging appears to affect the inactive and active X chromosomes equally. PMID- 10677327 TI - Age estimate of the N370S mutation causing Gaucher disease in Ashkenazi Jews and European populations: A reappraisal of haplotype data. AB - The N370S mutation at the GBA locus on human chromosome 1q21, which causes Gaucher disease (GD), has a high frequency in the Ashkenazim and is the second most-widespread GD mutation in the European non-Jewish population. A common ancient origin for the N370S mutation in the Ashkenazi Jewish and Spanish populations has been proposed on the basis of both a similar haplotype for associated markers and an age estimate that suggests that this mutation appeared several thousand years ago. However, a reappraisal of haplotype data, using the Risch formula properly along with a Luria-Delbruck setting of the genetic clock, allows identification of the likely origin of the N370S mutation in Ashkenazi Jews between the 11th and 13th centuries. This result is consistent with the estimated ages of other mutations that are frequent among Ashkenazim, with the exception of type II (Glu117Stop) factor XI deficiency, which is deemed to be >3000 years old, predating the separation of the Ashkenazi and Iraqi Jews. The present finding supports the hypothesis of a more recent origin for the N370S mutation and is consistent with both a founder chromosome transfer from Ashkenazim who assimilated in some European populations and a non-Jewish origin of the European N370S-bearing chromosomes. PMID- 10677328 TI - A new locus for generalized epilepsy with febrile seizures plus maps to chromosome 2. AB - Generalized epilepsy with febrile seizures plus (GEFS+) is a recently recognized but relatively common form of inherited childhood-onset epilepsy with heterogeneous epilepsy phenotypes. We genotyped 41 family members, including 21 affected individuals, to localize the gene causing epilepsy in a large family segregating an autosomal dominant form of GEFS+. A genomewide search examining 197 markers identified linkage of GEFS+ to chromosome 2, on the basis of an initial positive LOD score for marker D2S294 (Z=4.4, recombination fraction [straight theta] = 0). A total of 24 markers were tested on chromosome 2q, to define the smallest candidate region for GEFS+. The highest two-point LOD score (Zmax=5.29; straight theta=0) was obtained with marker D2S324. Critical recombination events mapped the GEFS+ gene to a 29-cM region flanked by markers D2S156 and D2S311, with the idiopathic generalized epilepsy locus thereby assigned to chromosome 2q23-q31. The existence of the heterogeneous epilepsy phenotypes in this kindred suggests that seizure predisposition determined by the GEFS+ gene on chromosome 2q could be modified by other genes and/or by environmental factors, to produce the different seizure types observed. PMID- 10677329 TI - A new locus for autosomal dominant pure spastic paraplegia, on chromosome 2q24 q34. AB - Hereditary spastic paraplegia (HSP) comprises a group of clinically and genetically heterogeneous disorders causing progressive spasticity and weakness of the lower limbs. We report a large family of French descent with autosomal dominant pure HSP. We excluded genetic linkage to the known loci causing HSP and performed a genomewide search. We found evidence for linkage of the disorder to polymorphic markers on chromosome 2q24-q34: a maximum LOD score of 3. 03 was obtained for marker D2S2318. By comparison with families having linkage to the major locus of pure autosomal dominant HSP (SPG4 on chromosome 2p), there were significantly more patients without Babinski signs, with increased reflexes in the upper limbs, and with severe functional handicaps. PMID- 10677330 TI - Absence of significant linkage between phonological coding dyslexia and chromosome 6p23-21.3, as determined by use of quantitative-trait methods: confirmation of qualitative analyses. AB - We recently reported the absence of significant linkage of phonological coding dyslexia (PCD) to chromosome 6p23-p21.3 in 79 families with at least two affected siblings, even though linkage of dyslexia to this region has been found in four other independent studies. Whereas, in our previous analyses, we used a qualitative (affected, unaffected, or uncertain) PCD phenotype, here we report a reanalysis of linkage to the chromosome 6p region, by use of four quantitative measures of reading disability: phonological awareness, phonological coding, spelling, and rapid-automatized-naming (RAN) speed. The phonological-coding and spelling measures were highly correlated with each other and with the qualitative PCD phenotype, whereas the phonological-awareness and RAN-speed measures were only moderately correlated with the other measures. Using two-point and multipoint quantitative-trait sib-pair linkage analyses and variance-components analyses, we were unable to detect significant evidence for a locus in the 6p23 p21.3 region influencing any of the quantitative reading measures, supporting our previous qualitative linkage results. The most likely explanation for our inability to detect linkage between dyslexia and this region is that families with subtypes of dyslexia linked to this region are underrepresented in our sample, because of either chance or varying ascertainment criteria. PMID- 10677331 TI - Chromosome 6p influences on different dyslexia-related cognitive processes: further confirmation. AB - In this study, which is a continuation and an extension of an earlier study, we enrolled two new families (N=31) and recruited more individuals from the previously ascertained families (N=56). The eight multiplex families (N=171) presented in this study were ascertained from a sample of adult probands whose childhood reading history is well documented through archival information. Six phenotypes were constructed to span a range of dyslexia-related cognitive processes. These phenotypes were (1) phonemic awareness (of spoken words); (2) phonological decoding (of printed nonwords); (3) rapid automatized naming (of colored squares or object drawings); (4) single-word reading (orally, of printed real words); (5) vocabulary; and (6) spelling (of dictated words). In addition, the diagnosis of lifelong dyslexia was established by clinical means. Genotyping was done with nine highly polymorphic markers from the 6p22.3-6p21.3 region. The results of two- and multipoint identity-by-descent and identity-by-state analyses supported the importance of a putative locus in the D6S464-D6S273 region for a number of dyslexia-related cognitive deficits. PMID- 10677332 TI - A third novel locus for primary autosomal recessive microcephaly maps to chromosome 9q34. AB - Primary autosomal recessive microcephaly is a clinical diagnosis of exclusion in an individual with a head circumference >/=4 SDs below the expected age-and-sex mean. There is associated moderate mental retardation, and neuroimaging shows a small but structurally normal cerebral cortex. The inheritance pattern in the majority of cases is considered to be autosomal recessive. Although genetic heterogeneity for this clinical phenotype had been expected, this has only recently been demonstrated, with the mapping of two loci for autosomal recessive primary microcephaly: MCPH1 at 8p and MCPH2 at 19q. We have studied a large multiaffected consanguineous pedigree, using a whole-genome search, and have identified a third locus, MCPH3 at 9q34. The minimal critical region is approximately 12 cM, being defined by the markers cen-D9S1872-D9S159-tel, with a maximum two-point LOD score of 3.76 (recombination fraction 0) observed for the marker D9S290. PMID- 10677333 TI - A locus for autosomal dominant "pure" hereditary spastic paraplegia maps to chromosome 19q13. AB - Genetic loci for autosomal dominant pure hereditary spastic paraplegia (ADPHSP) have been mapped to chromosomes 2p, 8q, 12q, 14q, and 15q. We undertook a genomewide linkage screen of a large family with ADPHSP, for which linkage at all previously identified ADPHSP loci was excluded. Analysis of markers on chromosome 19q gave a peak pairwise LOD score of 3.72 at D19S420, allowing assignment of a novel ADPHSP locus (which we have termed "SPG12") to this region. Haplotype construction and analysis of recombination events narrowed the SPG12 locus to a 16.1-cM region between markers D19S868 and D19S902. PMID- 10677334 TI - Vacuoliting megalencephalic leukoencephalopathy with subcortical cysts, mapped to chromosome 22qtel. AB - The leukodystrophies form a complex group of orphan genetic disorders that primarily affect myelin, the main constituent of the brain white matter. Among the leukodystrophies of undetermined etiology, a new clinical entity called "vacuoliting megalencephalic leukoencephalopathy" (VL) was recently recognized. VL is characterized by diffuse swelling of the white matter, large subcortical cysts, and megalencephaly with infantile onset. Family studies in several ethnic groups have suggested an autosomal recessive mode of inheritance. We mapped the VL gene to chromosome 22qtel, within a 3-cM linkage interval between markers D22S1161 and n66c4 (maximum LOD score 10.12 at recombination fraction.0, for marker n66c4; maximum multipoint LOD score 17 for this interval) by genome scan of 13 Turkish families. Linkage analysis under the genetic-heterogeneity hypothesis showed no genetic heterogeneity. No abnormalities were found in three tested candidate genes (fibulin-1 and glutathione S-transferases 1 and 2). PMID- 10677335 TI - Application and interpretation of transmission/disequilibrium tests: transmission of HLA-DQ haplotypes to unaffected siblings in 526 families with type 1 diabetes. AB - It is widely believed that, if a genetic marker shows a transmission distortion in patients by the transmission/disequilibrium test (TDT), then a transmission distortion in healthy siblings would be seen in the opposite direction. This is also the case in a complex disease. Furthermore, it has been suggested that replacing the McNemar statistics of the TDT with a test of heterogeneity between transmissions to affected and unaffected children could increase the power to detect disease association. To test these two hypotheses empirically, we analyzed the transmission of HLA-DQA1-DQB1 haplotypes in 526 Norwegian families with type 1 diabetic children and healthy siblings, since some DQA1-DQB1 haplotypes represent major genetic risk factors for type 1 diabetes. Despite the strong positive and negative disease associations with particular DQ haplotypes, we observed no significant deviation from 50% for transmission to healthy siblings. This could be explained by the low penetrance of susceptibility alleles, together with the fact that IDDM loci also harbor strongly protective alleles that can override the risk contributed by other loci. Our results suggest that, in genetically complex diseases, detectable distortion in transmission to healthy siblings should not be expected. Furthermore, the original TDT seems more powerful than a heterogeneity test. PMID- 10677336 TI - The 1298(A-->C) mutation of methylenetetrahydrofolate reductase should be designated to the 1289 position of the gene. PMID- 10677338 TI - Special oversight groups to add protections for population-based repository samples. PMID- 10677339 TI - Letter to human genetics journals. PMID- 10677340 TI - On the preparation of beta-haematin. AB - Synthetic beta-haematin is considered to be identical, or at least very similar to, purified malaria pigment. Methods for its preparation use ferriprotoporphyrin IX at acid pH in the presence of acetic acid at different concentrations and degrees of ionization, elevated temperatures and long reaction times. Here we show that certain widely used reaction conditions, involving very high concentrations of acetic acid/acetate mixtures, do not produce substantial amounts of polymeric beta-haematin on immediate isolation of the reaction products, but only during prolonged drying of the products at 37 degrees C after washing with water. Alternative, more convenient methods of preparation of pure beta-haematin are suggested. PMID- 10677341 TI - The ShBle resistance determinant from Streptoalloteichus hindustanus is expressed in Haloferax volcanii and confers resistance to bleomycin. AB - We have designed a gene cassette for expression of the bleomycin-resistance protein from Streptoalloteichus hindustanus (ShBle) in the extremely halophilic archaeon Haloferax volcanii, and shown that transformed haloarchaea are resistant to bleomycin. Recombinant ShBle was purified by a one-step affinity chromatography procedure as a correctly folded, dimeric protein. ShBle thus provides a useful haloarchaeal selectable marker and represents the first non halophilic and soluble heterologous protein to be expressed in the Haloarchaea. PMID- 10677342 TI - Mechanism of thermal denaturation of maltodextrin phosphorylase from Escherichia coli. AB - Maltodextrin phosphorylase from Escherichia coli (MalP) is a dimeric protein in which each approximately 90-kDa subunit contains active-site pyridoxal 5' phosphate. To unravel factors contributing to the stability of MalP, thermal denaturations of wild-type MalP and a thermostable active-site mutant (Asn-133- >Ala) were compared by monitoring enzyme activity, cofactor dissociation, secondary structure content and aggregation. Small structural transitions of MalP are shown by Fourier-transform infrared spectroscopy to take place at approximately 45 degrees C. They are manifested by slight increases in unordered structure and (1)H/(2)H exchange, and reflect reversible inactivation of MalP. Aggregation of the MalP dimer is triggered by these conformational changes and starts at approximately 45 degrees C without prior release into solution of pyridoxal 5'-phosphate. It is driven by electrostatic rather than hydrophobic interactions between MalP dimers, and leads to irreversible inactivation of the enzyme. Aggregation is inhibited efficiently and specifically by oxyanions such as phosphate, and AMP which therefore, stabilize MalP against the irreversible denaturation step at 45 degrees C. Melting of the secondary structure in soluble and aggregated MalP takes place at much higher temperatures of approx. 58 and 67 degrees C, respectively. Replacement of Asn-133 by Ala does not change the mechanism of thermal denaturation, but leads to a shift of the entire pathway to a approximately 15 degrees C higher value on the temperature scale. Apart from greater stability, the Asn-133-->Ala mutant shows a 2-fold smaller turnover number and a 4.6-fold smaller energy of activation than wild-type MalP, probably indicating that the site-specific replacement of Asn-133 brings about a greater rigidity of the active-site environment of the enzyme. A structure-based model is proposed which explains the stabilizing interaction between MalP and oxyanions, or AMP. PMID- 10677343 TI - Some polyphenols inhibit the formation of pentyl radical and octanoic acid radical in the reaction mixture of linoleic acid hydroperoxide with ferrous ions. AB - Effects of some polyphenols and their related compounds (chlorogenic acid, caffeic acid, quinic acid, ferulic acid, gallic acid, D-(+)-catechin, D-(-) catechin, 4-hydroxy-3-methoxybenzoic acid, salicylic acid, L-dopa, dopamine, L adrenaline, L-noradrenaline, o-dihydroxybenzene, m-dihydroxybenzene, and p dihydroxybenzene) on the formation of 13-hydroperoxide octadecadienoic (13-HPODE) acid-derived radicals (pentyl radical and octanoic acid radical) were examined. The ESR spin trapping showed that chlorogenic acid, caffeic acid, gallic acid, D (+)-catechin, D-(-)-catechin, L-dopa, dopamine, L-adrenaline, L-noradrenaline, and o-dihydroxybenzene inhibited the overall formation of 13-HPODE acid-derived radicals in the reaction mixture of 13-HPODE with ferrous ions. The ESR peak heights of alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN)/13-HPODE derived radical adducts decreased to 46+/-4% (chlorogenic acid), 54+/-2% (caffeic acid), 49+/-2% (gallic acid), 55+/-1% [D-(+)-catechin], 60+/-3% [D-(-)-catechin], 42+/-1% (L-dopa), 30+/-2% (dopamine), 49+/-2% (L-adrenaline), 24+/-2% (L noradrenaline), and 54+/-5% (o-dihydroxybenzene) of the control, respectively. The high performance liquid chromatography-electron spin resonance (HPLC-ESR) and high performance liquid chromatography-electron spin resonance-mass spectrometries (HPLC-ESR-MS) showed that caffeic acid inhibited the formation of octanoic acid radical and pentyl radical to 42+/-2% and 52+/-7% of the control, respectively. On the other hand, the polyphenols and their related compounds had few inhibitory effects on the radical formation in the presence of EDTA. Visible absorbance measurement revealed that all the polyphenols exhibiting the inhibitory effect chelate ferrous ions. Above results indicated that the chelation of ferrous ion is essential to the inhibitory effects of the polyphenols. PMID- 10677344 TI - Ca2+ and calmodulin differentially modulate myo-inositol 1,4, 5-trisphosphate (IP3)-binding to the recombinant ligand-binding domains of the various IP3 receptor isoforms. AB - We have expressed the N-terminal 581 amino acids of type 1 myo-inositol 1,4,5 trisphosphate receptor (IP(3)R1), IP(3)R2 and IP(3)R3 as recombinant proteins [ligand-binding site 1 (lbs-1), lbs-2, lbs-3] in the soluble fraction of Escherichia coli. These recombinant proteins contain the complete IP(3)-binding domain and bound IP(3) and adenophostin A with high affinity. Ca(2+) and calmodulin were previously found to maximally inhibit IP(3) binding to lbs-1 by 42+/-6 and 43+/-6% respectively, and with an IC(50) of approx. 200 nM and 3 microM respectively [Sipma, De Smet, Sienaert, Vanlingen, Missiaen, Parys and De Smedt (1999) J. Biol. Chem. 274, 12157-12562]. We now report that Ca(2+) inhibited IP(3) binding to lbs-3 with an IC(50) of approx. 700 nM (37+/-4% inhibition at 5 microM Ca(2+)), while IP(3) binding to lbs-2 was not affected by increasing [Ca(2+)] from 100 nM to 25 microM. Calmodulin (10 microM) inhibited IP(3) binding to lbs-3 by 37+/-4%, while IP(3) binding to lbs-2 was inhibited by only 11+/-2%. The inhibition of IP(3) binding to lbs-3 by calmodulin was dose dependent (IC(50) approximately 2 microM). We conclude that the IP(3)-binding domains of the various IP(3)R isoforms differ in binding characteristics for IP(3) and adenophostin A, and are differentially modulated by Ca(2+) and calmodulin, suggesting that the various IP(3)R isoforms can have different intracellular functions. PMID- 10677345 TI - Pancreatic eukaryotic initiation factor-2alpha kinase (PEK) homologues in humans, Drosophila melanogaster and Caenorhabditis elegans that mediate translational control in response to endoplasmic reticulum stress. AB - In response to different cellular stresses, a family of protein kinases regulates translation by phosphorylation of the alpha subunit of eukaryotic initiation factor-2 (eIF-2alpha). Recently, we identified a new family member, pancreatic eIF-2alpha kinase (PEK) from rat pancreas. PEK, also referred to as RNA-dependent protein kinase (PKR)-like endoplasmic reticulum (ER) kinase (PERK) is a transmembrane protein implicated in translational control in response to stresses that impair protein folding in the ER. In this study, we identified and characterized PEK homologues from humans, Drosophila melanogaster and Caenorhabditis elegans. Expression of human PEK mRNA was found in over 50 different tissues examined, with highest levels in secretory tissues. In mammalian cells subjected to ER stress, we found that elevated eIF-2alpha phosphorylation was coincident with increased PEK autophosphorylation and eIF 2alpha kinase activity. Activation of PEK was abolished by deletion of PEK N terminal sequences located in the ER lumen. To address the role of C. elegans PEK in translational control, we expressed this kinase in yeast and found that it inhibits growth by hyperphosphorylation of eIF-2alpha and inhibition of eIF-2B. Furthermore, we found that vaccinia virus K3L protein, an inhibitor of the eIF 2alpha kinase PKR involved in an anti-viral defence pathway, also reduced PEK activity. These results suggest that decreased translation initiation by PEK during ER stress may provide the cell with an opportunity to remedy the folding problem prior to introducing newly synthesized proteins into the secretory pathway. PMID- 10677346 TI - Target site search and effective inhibition of leukaemic cell growth by a covalently closed multiple anti-sense oligonucleotide to c-myb. AB - Systematic secondary structure simulation of a target mRNA sequence is shown to be effective for locating a good anti-sense target site. Multiple selected anti sense sequences were placed in a single molecule. The anti-sense oligonucleotide (oligo) was covalently closed to avoid exonuclease activities and was designated CMAS (covalently closed multiple anti-sense)-oligo. CMAS-oligo was found to be stable, largely preserving its structural integrity after 24 h of incubation in the presence of either exonuclease III or serum. When human c-myb mRNA was targeted by the c-myb CMAS-oligo, expression of the gene was completely abolished. Further, tumour cell growth was inhibited by 82+/-3% as determined by an MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay and by 90+/-1% by [(3)H]thymidine incorporation. When a leukaemic cell line K562 was treated with CMAS-oligo, colony formation on soft agarose was also decreased by 93%. In contrast, treatment with a scrambled control oligo did not significantly inhibit leukaemic cell growth. These results suggest that a rational target site search is possible for an anti-sense oligo and that CMAS oligo can be employed as an effective anti-sense agent with enhanced stability. PMID- 10677347 TI - HIV-2 protease is inactivated after oxidation at the dimer interface and activity can be partly restored with methionine sulphoxide reductase. AB - Human immunodeficiency viruses encode a homodimeric protease that is essential for the production of infectious virus. Previous studies have shown that HIV-1 protease is susceptible to oxidative inactivation at the dimer interface at Cys 95, a process that can be reversed both chemically and enzymically. Here we demonstrate a related yet distinct mechanism of reversible inactivation of the HIV-2 protease. Exposure of the HIV-2 protease to H(2)O(2) resulted in conversion of the two methionine residues (Met-76 and Met-95) to methionine sulphoxide as determined by amino acid analysis and mass spectrometry. This oxidation completely inactivated protease activity. However, the activity could be restored (up to 40%) after exposure of the oxidized protease to methionine sulphoxide reductase. This treatment resulted in the reduction of methionine sulphoxide 95 but not methionine sulphoxide 76 to methionine, as determined by peptide mapping/mass spectrometry. We also found that exposure of immature HIV-2 particles to H(2)O(2) led to the inhibition of polyprotein processing in maturing virus particles comparable to that demonstrated for HIV-1 particles. Thus oxidative inactivation of the HIV protease in vitro and in maturing viral particles is not restricted to the type 1 proteases. These studies indicate that two distinct retroviral proteases are susceptible to inactivation after a very minor modification at residue 95 of the dimer interface and suggest that the dimer interface might be a viable target for the development of novel protease inhibitors. PMID- 10677348 TI - Inhibition of atrial natriuretic peptide (ANP) C receptor expression by antisense oligodeoxynucleotides in A10 vascular smooth-muscle cells is associated with attenuation of ANP-C-receptor-mediated inhibition of adenylyl cyclase. AB - Atrial natriuretic peptide (ANP) mediates a variety of physiological effects through its interaction with ANP-A, ANP-B or ANP-C receptors. However, controversies exist regarding the involvement of ANP-C receptor and adenylyl cyclase/cAMP signal-transduction systems to which these receptors are coupled in mediating these responses. In the present studies, we have employed an antisense approach to eliminate the ANP-C receptor and to examine the effect of this elimination on adenylyl cyclase inhibition. An 18-mer antisense phosphorothioate oligodeoxynucleotide (OH-2) targeted at the initiation codon of the ANP-C receptor was used to examine its effects on the expression of the ANP-C receptor and ANP-C-receptor-mediated inhibition of adenylyl cyclase in vascular smooth muscle cells (A10). Treatment of the cells with antisense oligonucleotide resulted in complete attenuation of C-ANP(4-23) [des(Gln(18), Ser(19), Gln(20), Leu(21), Gly(22))ANP(4-23)-NH(2)]-mediated inhibition of adenylyl cyclase, whereas sense and missense oligomers did not affect the inhibition of adenylyl cyclase by C-ANP(4-23). In addition, the stimulatory effects of guanine nucleotides, isoproterenol, sodium fluoride and forskolin as well as the inhibitory effects of angiotensin II on adenylyl cyclase were not affected by antisense-oligonucleotide treatment. The attenuation of C-ANP(4-23)-mediated inhibition of adenylyl cyclase by antisense oligonucleotide was dose- and time dependent. A complete attenuation of ANP-C-receptor-mediated inhibition of adenylyl cyclase was observed at 2.5 microM. In addition, treatment of the cells with antisense oligonucleotide and not with sense or missense oligomers resulted in the inhibition of the levels of ANP-C-receptor protein and mRNA as determined by immunoblotting and Northern blotting using antisera against the ANP-C receptor and a cDNA probe of the ANP-C receptor respectively. On the other hand, ANP-A/B receptor-mediated increases in cGMP levels were not inhibited by antisense oligonucleotide treatment. Our results demonstrate conclusively that the elimination of ANP-C receptor by antisense oligonucleotide attenuates ANP-induced inhibition of adenylyl cyclase and provide evidence that antisense oligonucleotide of the ANP-C receptor may serve as a useful pharmacological tool to elucidate the physiological functions of the ANP-C receptor. PMID- 10677349 TI - Actin filaments play a critical role in insulin-induced exocytotic recruitment but not in endocytosis of GLUT4 in isolated rat adipocytes. AB - Actin-based cytoskeletons have been implicated in insulin-stimulated glucose transport and translocation of the insulin-regulated glucose transporter, GLUT4, from the intracellular pool to the plasma membrane. However, most previous studies were done using adherent cell systems such as L6 myotubes and 3T3-L1 adipocytes, and very little information is available on the significance of the actin filaments to the insulin action in isolated adipocytes, a widely used experimental system. In the present study, we investigated the physiological role of actin filaments in the subcellular trafficking of GLUT4 in isolated rat adipocytes. We first compared the effects of two actin-disrupting reagents, latrunculin A and cytochalasin D, on the organization of the actin filaments as well as on the insulin action on glucose transport by laser confocal microscopy combined with biochemical analysis of the insulin action. Treatment of the cells with latrunculin A induced dose- and time-dependent disappearance of the filamentous actin, which correlated very well with inhibition of the insulin effect on glucose transport. Although cytochalasin D at 50 microM significantly inhibited insulin-stimulated glucose transport, it was not effective in disassembly of the actin filaments; rather, many intense punctate signals were observed in cytochalasin D-treated cells. In the actin-disrupted adipocytes treated with latrunculin A, insulin-induced GLUT4 translocation was inhibited completely. In addition, latrunculin A remarkably inhibited both insulin-induced glucose transport and GLUT4 translocation in the presense of D(k)-(62-85), a potent inhibitor of GLUT4 endocytosis, suggesting that intactness of the actin filaments was necessary for insulin-induced exocytosis of the GLUT4-containing vesicles. On the other hand, latrunculin A showed little inhibitory effect on either endocytosis of the trypsin-cleaved 35-kDa fragment of GLUT4 or decay of the glucose transport activity after addition of wortmannin in insulin-stimulated cells. The results of our experiment show clearly that, in rat adipocytes, (i) latrunculin A may be a more suitable tool than cytochalasin D for disruption of actin filaments, and (ii) actin filaments play a crucial role in exocytotic recruitment of GLUT4 to the plasma membrane from the intracellular pool, but not in its endocytosis. PMID- 10677350 TI - Zinc-regulated ubiquitin conjugation signals endocytosis of the yeast ZRT1 zinc transporter. AB - The yeast ZRT1 zinc transporter is regulated by zinc at both transcriptional and post-translational levels. At the post-translational level, zinc inactivates ZRT1 by inducing the removal of the protein from the plasma membrane by endocytosis. The zinc transporter is subsequently degraded in the vacuole. This regulatory system allows for the rapid shut off of zinc uptake activity in cells exposed to high zinc concentrations, thereby preventing overaccumulation of this potentially toxic metal. In this report, we examine the role of ubiquitin conjugation in this process. First, we show that ZRT1 is ubiquitinated shortly after zinc treatment and before endocytosis. Secondly, mutations in various components of the ubiquitin conjugation pathway, specifically the RSP5 ubiquitin-protein ligase and the UBC4 and UBC5 ubiquitin conjugating enzymes, inhibit both ubiquitination and endocytosis. Finally, mutation of a specific lysine residue in ZRT1 blocks both ubiquitination and endocytosis. This critical lysine, Lys-195, is located in a cytoplasmic loop region of the protein and may be the residue to which ubiquitin is attached. These results demonstrate that ubiquitin conjugation is a critical step in the signal transduction pathway that controls the rate of ZRT1 endocytosis in response to zinc. PMID- 10677351 TI - Down-regulation of cyclic-nucleotide phosphodiesterase 3B in 3T3-L1 adipocytes induced by tumour necrosis factor alpha and cAMP. AB - We have used murine 3T3-L1 cells, which differentiate in culture and acquire morphological and biochemical features of mature adipocytes, as a model for studying the expression of cyclic-nucleotide phosphodiesterase (PDE) 3B activity, protein and mRNA during differentiation and during long-term treatment of the cells with tumour necrosis factor alpha (TNF-alpha), a cytokine associated with insulin resistance, and a cAMP analogue, N(6),2'-O-dibutyryl cAMP (dbcAMP). PDE3B activity, protein and mRNA could be detected 4 days after the initiation of differentiation of 3T3-L1 preadipocytes. Treatment of 3T3-L1 adipocytes with 10 ng/ml TNF-alpha for 24 h produced a maximal (50%) decrease in PDE3B activity, protein and mRNA, which was well correlated with both activation of protein kinase A (PKA) and stimulation of lipolysis, presumably reflecting an increase in intracellular cAMP concentration. To investigate the effect of cAMP on PDE3B we treated 3T3-L1 adipocytes with dbcAMP. After 4 h with 0.5 mM dbcAMP, PDE3B activity was decreased by 80%, which was also correlated with a decrease in PDE3B protein and mRNA. This effect was abolished in the presence of N-[2 (bromocinnamylamino)ethyl]-5-isoquinolinesulphonamide] (H-89), a specific PKA inhibitor. We conclude that the lipolytic effect of TNF-alpha involves the down regulation of PDE3B, which is associated with increased activation of PKA, presumably owing to increased levels of cAMP. In addition, the PKA activation induced by dbcAMP resulted in the down-regulation of PDE3B. These results, which suggest that PDE3B is a novel target for long-term regulation by TNF-alpha and cAMP, could contribute to the understanding of the mechanisms of insulin resistance. PMID- 10677352 TI - Reagent or myeloperoxidase-generated hypochlorite affects discrete regions in lipid-free and lipid-associated human apolipoprotein A-I. AB - We have previously shown that the modification of high-density lipoprotein subclass 3 (HDL(3)) by HOCl transformed an anti-atherogenic lipoprotein into a high-uptake form for macrophages and caused a significant impairment of cholesterol efflux capacity [Panzenboeck, Raitmayer, Reicher, Lindner, Glatter, Malle and Sattler (1997) J. Biol. Chem. 272, 29711-29720]. To elucidate the consequences of treatment with OCl(-) on distinct regions in apolipoprotein A-I (apo A-I), lipid-free and lipid-associated apo A-I were modified with increasing molar ratios of NaOCl or HOCl generated by the myeloperoxidase/H(2)O(2)/Cl(-) system. CD analysis revealed a pronounced decrease in alpha-helicity for lipid free apo A-I modified by NaOCl, whereas lipid-associated apo A-I was less affected. The modification of apo A-I by NaOCl (molar oxidant-to-lipoprotein ratio 6:1) resulted in the formation of two distinct oxidized forms of apo A-I with molecular masses 32 or 48 atomic mass units (a.m.u.) higher than that of native apo A-I, indicating the addition of two or three oxygen atoms to the native protein. HPLC analysis of tryptic digests obtained from lipid-free and lipid-associated apo A-I modified with increasing oxidant-to-apolipoprotein molar ratios revealed a concentration-dependent modification of apo A-I: at a low molar oxidant-to-lipoprotein ratio (5:1) the peaks corresponding to the methionine containing tryptic peptides T11 (residues 84-88), T16 (residues 108-116) and T22 (residues 141-149), located in the central region of apo A-I, disappeared. Their loss was accompanied by the formation of three oxidation products with a molecular mass 16 a.m.u. higher than that of the native peptides. This indicates the addition of oxygen, most probably caused by the oxidation of Met(86), Met(112) and Met(148) to the corresponding methionine sulphoxides. At a molar NaOCl-to-apo A-I ratio of 10:1 the disappearance of peptides T1 (residues 1-10), T7 (residues 46-59) and T9 (residues 62-77) was accompanied by the occurrence of new peaks 33.5 and 33.1 a.m.u. higher than those of the native peptides. Amino acid analyses of peptides T7 and T9 after modification with NaOCl confirmed that Phe(57) and Phe(71) were primary targets for oxidation by HOCl. GLC-MS analysis of hydrolysates obtained from OCl(-)-modified T7, T9, apo A-I and HDL(3) confirmed that Phe residues are an early target for OCl(-) modification. At molar NaOCl-to-apo A-I ratios of 25:1, the peak areas of peptides T31 (residues 189 195) and T32 (residues 196-206) decreased markedly. Most importantly, incubation of apo A-I with the myeloperoxidase/H(2)O(2)/Cl(-) system (the source of HOCl in vivo) resulted in almost identical modification patterns to those observed with reagent NaOCl. PMID- 10677353 TI - Enzyme activity and dynamics: xylanase activity in the absence of fast anharmonic dynamics. AB - The activity and dynamics of a simple, single subunit enzyme, the xylanase from Thermotoga maritima strain Fj SS3B.1 have been measured under similar conditions, from -70 to +10 degrees C. The internal motions of the enzyme, as evidenced by neutron scattering, undergo a sharp transition within this temperature range; they show no evidence for picosecond-timescale anharmonic behaviour (e.g. local diffusive motions or jumps between alternative conformations) at temperatures below -50 degrees C, whereas these motions are strongly activated at higher temperatures. The activity follows Arrhenius behaviour over the whole of the temperature range investigated, -70 to +10 degrees C. The results indicate that a temperature range exists over which the enzyme rate-limiting step is independent of fast anharmonic dynamics. PMID- 10677354 TI - Stimulation of cleavage of membrane proteins by calmodulin inhibitors. AB - The ectodomain of several membrane-bound proteins can be shed by proteolytic cleavage. The activity of the proteases involved in shedding is highly regulated by several intracellular second messenger pathways, such as protein kinase C (PKC) and intracellular Ca(2+). Recently, the shedding of the adhesion molecule L selectin has been shown to be regulated by the interaction of calmodulin (CaM) with the cytosolic tail of L-selectin. Prevention of CaM-L-selectin interaction by CaM inhibitors or mutation of a CaM binding site in L-selectin induced L selectin ectodomain shedding. Whether this action of CaM inhibitors also affects other membrane-bound proteins is not known. In the present paper we show that CaM inhibitors also stimulate the cleavage of several other transmembrane proteins, such as the membrane-bound growth factor precursors pro-transforming growth factor-alpha and pro-neuregulin-alpha2c, the receptor tyrosine kinase, TrkA, and the beta-amyloid precursor protein. Cleavage induced by CaM inhibitors was a rapid event, and resulted from the activation of a mechanism that was independent of PKC or intracellular Ca(2+) increases, but was highly sensitive to hydroxamic acid-based metalloprotease inhibitors. Mutational analysis of the intracellular domain of the TrkA receptor indicated that CaM inhibitors may stimulate membrane protein ectodomain cleavage by mechanisms independent of CaM-substrate interaction. PMID- 10677355 TI - Stress-induced generation of N-acylethanolamines in mouse epidermal JB6 P+ cells. AB - It has long been known that N-acylethanolamine phospholipids [N acylphosphatidylethanolamine (N-acyl PE)] and N-acylethanolamines (NAEs) accumulate in mammalian tissues undergoing degenerative membrane changes associated with necrosis. Here we studied the effects of stress factors (UVB irradiation and serum deprivation) on the endogenous levels of N-acyl PE and NAE in mouse epidermal JB6 P(+) cells. We found that 16:0, 18:0, 18:1,n-9 and 18:1,n 7 are the predominant amide-linked fatty acids in both N-acyl PE and NAE in these cells. UVB irradiation and serum deprivation resulted in significantly increased levels of N-acyl PE and NAE, especially 18:1, n-9 N-acyl PE and NAE. UVB challenge increased the cellular content of anandamide (20:4,n-6 NAE), but this increase was the lowest of all NAEs measured. Serum deprivation resulted in a decreased cellular anandamide level, as well as a decrease in 20:4,n-6 N-acyl PE. Interestingly, the replacement of serum-free medium with medium containing 5% (v/v) fetal calf serum after 36 h of serum deprivation restored N-acyl PE and NAE levels almost completely within 4-8 h. These data suggest the involvement of N acyl PE and NAE in cellular responses to stress. PMID- 10677356 TI - Ferredoxin III of Desulfovibrio africanus: sequencing of the native gene and characterization of a histidine-tagged form. AB - Desulfovibrio africanus ferredoxin III (Da FdIII) contains one [4Fe-4S](2+/1+) cluster and one [3Fe-4S](1+/0) cluster, bound by seven Cys residues, in which the [3Fe-4S] cluster is co-ordinated by the unusual sequence, Cys(11)-Xaa-Xaa-Asp(14) Xaa-Xaa-Cys(17)-Xaa(n)-Cys(51)-Glu. The [3Fe-4S] core of this ferredoxin is so far unique in showing rapid bi-directional [3Fe-4S]<-->[4Fe-4S] cluster interconversion with a wide range of metal ions. In order to obtain protein for mutagenesis studies Da FdIII has been cloned, sequenced, and expressed as a hexa histidine tagged (ht) polypeptide in Escherichia coli strain BL21(DE3) pLysS. Expression of ht Da FdIII, whether translated from a synthetic gene (pJB10) or from the native nucleotide sequence (pJB11), occurred at similar levels (approx. 6 mg.l(-1)), but without incorporation of metal clusters. The nucleotide sequence confirms the protein sequence reported previously [Bovier-Lapierre, Bruschi, Bonicel and Hatchikian (1987) Biochim. Biophys. Acta 913, 20-26]. Cluster incorporation was achieved using FeCl(3) together with cysteine sulphur transferase, NifS, plus cysteine to generate low levels of sulphide ions. Absorption and EPR spectroscopy show that both [3Fe-4S] and [4Fe-4S] clusters are correctly inserted. Thin-film electrochemistry provides evidence that the [3Fe 4S] cluster undergoes reversible cluster transformation in the presence of Fe(II) and Zn(II) ions with properties identical to the native protein. Nevertheless the protein has lower stability than native Da FdIII during chromatography. The one dimensional 600 MHz NMR spectrum of the apoprotein indicates an unstructured protein with random coil chemical shifts whereas spectra of the reconstituted ht protein show secondary structural elements and 18 peaks shifted downfield of 9.6 p.p.m. The spectra are unique but have similarities with the shift patterns seen with 7Fe Desulfurolobus ambivalens Fd. The ht does not affect iron-sulphur cluster incorporation, but NMR evidence suggests that excess Fe binds to the tag. This may account for the lower stability of the ht compared with the native protein. PMID- 10677357 TI - Estimation of systolic and diastolic free intracellular Ca2+ by titration of Ca2+ buffering in the ferret heart. AB - Spectroscopic Ca(2+)-indicators are thought to report values of free intracellular Ca(2+) concentration ([Ca(2+)](i)) that may differ from unperturbed values because they add to the buffering capacity of the tissue. To check this for the heart we have synthesized a new (19)F-labelled NMR Ca(2+) indicator, 1, 2 bis-[2-bis(carboxymethyl)amino-4,5-difluorophenoxy]ethane ('4, 5FBAPTA'), with a low affinity (K(d) 2950 nM). The new indicator and four previously described (19)F-NMR Ca(2+) indicators 1,2-bis-[2-(1 - carboxyethyl)(carboxymethyl)amino - 5 - fluorophenoxy]ethane ('DiMe-5FBAPTA'), 1, 2-bis-[2-(1 carboxyethyl)(carboxymethyl)amino-4-fluorophenoxy]ethane ('DiMe-4FBAPTA'), 1, 2 bis-[2-bis(carboxymethyl)amino-5-fluorophenoxy]ethane ('5FBAPTA') and 1, 2-bis-[2 bis(carboxymethyl)amino-5-fluoro-4-methylphenoxy]ethane ('MFBAPTA'), with dissociation constants for Ca(2+) ranging from 46 to 537 nM, have been used to measure [Ca(2+)](i), over the range from less than 100 nM to more than 3 microM, in Langendorff-perfused ferret hearts (30 degrees C, pH 7.4, paced at 1.0 Hz) by (19)F-NMR spectroscopy. Loading hearts with indicators resulted in buffering of the Ca(2+) transient. The measured end-diastolic and peak-systolic [Ca(2+)](i) were both positively correlated with indicator K(d). The positive correlations between indicator K(d) and the measured end-diastolic and peak-systolic [Ca(2+)](i) were used to estimate the unperturbed end-diastolic and peak-systolic [Ca(2+)](i) by extrapolation to K(d)=0 (diastolic) and to K(d)=infinity (systolic) respectively. The extrapolated values in the intact beating heart were 161 nM for end-diastolic [Ca(2+)](i) and 2650 nM for peak-systolic [Ca(2+)](i), which agree well with values determined from single cells and muscle strips. PMID- 10677358 TI - Ca2+ buffering in the heart: Ca2+ binding to and activation of cardiac myofibrils. AB - The measurement of cardiac Ca(2+) transients using spectroscopic Ca(2+) indicators is significantly affected by the buffering properties of the indicators. The aim of the present study was to construct a model of cardiac Ca(2+) buffering that satisfied the kinetic constraints imposed by the maximum attainable rates of cardiac contraction and relaxation on the Ca(2+) dissociation rate constants and which would account for the observed effects of (19)F-NMR indicators on the cardiac Ca(2+) transient in the Langendorff-perfused ferret heart. It is generally assumed that the Ca(2+) dependency of myofibril activation in cardiac myocytes is mediated by a single Ca(2+)-binding site on troponin C. A model based on 1:1 Ca(2+) binding to the myofilaments, however, was unable to reproduce our experimental data, but a model in which we assumed ATP-dependent co operative Ca(2+) binding to the myofilaments was able to reproduce these data. This model was used to calculate the concentration and dissociation constant of the ATP-independent myofilament Ca(2+) binding, giving 58 and 2.0 microM respectively. In addition to reproducing our experimental data on the concentration of free Ca(2+) ions in the cytoplasm ([Ca(2+)](i)), the resulting Ca(2+) and ATP affinities given by fitting of the model also provided good predictions of the Ca(2+) dependence of the myofibrillar ATPase activity measured under in vitro conditions. Solutions to the model also indicate that the Ca(2+) mobilized during each beat remains unchanged in the presence of the additional buffering load from Ca(2+) indicators. The new model was used to estimate the extent of perturbation of the Ca(2+) transient caused by different concentrations of indicators. As little as 10 microM of a Ca(2+) indicator with a dissociation constant of 200 nM will cause a 20% reduction in peak-systolic [Ca(2+)](i) and 30 microM will cause approx. 50% reduction in the peak-systolic [Ca(2+)](i) in a heart paced at 1.0 Hz. PMID- 10677359 TI - Thyroxine regulation of monolysocardiolipin acyltransferase activity in rat heart. AB - Treatment of rats with thyroxine has been shown to elevate the biosynthesis and content of cardiolipin in the heart [Cao, Cheng, Angel and Hatch (1995) Biochim. Biophys. Acta 1256, 241-244]. Treatment with thyroxine resulted in a 1.8-fold increase (P<0.025) in [1-(14)C]linoleate and a 1.7-fold increase (P<0.025) in [1 (14)C]oleate incorporated into cardiolipin in perfused hearts, compared with controls. The mechanism for the elevation in incorporation of unsaturated fatty acids into cardiolipin was a 1. 6-fold (P<0.025) increase in mitochondrial monolysocardiolipin acyltransferase activity. The results demonstrate that the acylation of cardiac monolysocardiolipin is regulated by thyroid hormone. Thus an elevation in cardiolipin biosynthesis is accompanied by an elevation in monolysocardiolipin acyltransferase activity to maintain the appropriate molecular species composition of cardiolipin in the cardiac mitochondrial membrane. We postulate that monolysocardiolipin acyltransferase might be a rate limiting enzyme for the molecular remodelling of cardiolipin in the heart. PMID- 10677360 TI - Continuous exposure to high concentrations of nitric oxide leads to persistent inhibition of oxygen consumption by J774 cells as well as extraction of oxygen by the extracellular medium. AB - Nitric oxide (NO) plays a key role in many physiological and pathophysiological events, including the control of cell respiration. Both reversible and irreversible inhibition of mitochondrial respiration have been reported following the generation of NO by cells. We have exposed the murine macrophage cell line J774 to high concentrations of NO, such as are generated in some pathological conditions, and determined their effect on oxygen consumption. We observed a persistent inhibition of respiration which was due to a redox-dependent, progressive inhibition of complex I activity. No other enzyme of the respiratory chain was inhibited in this way. At the same time, we detected a paradoxical removal of oxygen by the extracellular medium. This removal was due to a chemical interaction between dissolved oxygen and NO-related species released from cells exposed to NO. A similar removal of oxygen by the cell supernatant also occurred following activation of cells with cytokines and bacterial products. Thus, the amounts of NO generated during pathological conditions may contribute to tissue hypoxia both by inhibiting cell respiration and by promoting removal of oxygen from the extracellular medium. PMID- 10677361 TI - Investigation of the mechanism by which glucose analogues cause translocation of glucokinase in hepatocytes: evidence for two glucose binding sites. AB - Glucokinase translocates between the cytoplasm and nucleus of hepatocytes where it is bound to a 68 kDa protein. The mechanism by which glucose induces translocation of glucokinase from the nucleus was investigated using glucose analogues that are not phosphorylated by glucokinase. There was strong synergism on glucokinase translocation between effects of glucose analogues (glucosamine, 5 thioglucose, mannoheptulose) and sorbitol, a precursor of fructose 1-phosphate. In the absence of glucose or glucose analogues, sorbitol had a smaller effect than glucose on translocation. However, sorbitol potentiated the effects of glucose analogues. In the absence of sorbitol the effect of glucose on glucokinase translocation is sigmoidal with a Hill coefficient of 1.9 suggesting involvement of two glucose-binding sites. The effects of glucosamine and 5 thioglucose were also sigmoidal but with lower Hill Coefficients. In the presence of sorbitol, the effects of glucose, glucosamine and 5-thioglucose were hyperbolic. Mannoheptulose, unlike the other glucose analogues, had a hyperbolic effect on glucokinase translocation in the absence of sorbitol suggesting interaction with one site and was synergistic rather than competitive with glucose. The results favour a two-site model for glucokinase translocation involving either two glucose-binding sites or one binding-site for glucose and one for fructose 1-phosphate. The glucose analogues differed in their effects on the kinetics of purified glucokinase. Mannoheptulose caused the greatest decrease in co-operativity of glucokinase for glucose whereas N-acetylglucosamine had the smallest effect. The anomalous effects of mannoheptulose on glucokinase translocation and on the kinetics of purified glucokinase could be explained by a second glucose-binding site on glucokinase. PMID- 10677362 TI - Identification and expression of an allergen Asp f 13 from Aspergillus fumigatus and epitope mapping using human IgE antibodies and rabbit polyclonal antibodies. AB - The Aspergillus genus of fungi is known to be one of the most prevalent aeroallergens. On two-dimensional immunoblotting using patients' sera containing IgE specific for Asp f 13, an allergen with a molecular mass of 33 kDa and a pI of 6.2 was identified. This allergen was also present in A. fumigatus culture filtrates. Furthermore, the sequence of the Asp f 13 cDNA was identical to that for alkaline protease isolated from A. fumigatus and showed 42-49% identity of amino acids with two proteases from P. cyclopium and T. album and with the Pen c 1 allergen from P. citrinum. Asp f 13 coding sequences were expressed in Escherichia coli as a [His](6)-tagged fusion protein which was purified by Ni(2+) chelate affinity chromatography. Recombinant Asp f 13 was recognized by rabbit polyclonal antibodies against Asp f 13 and by IgE antibodies from subject allergic to A. fumigatus. To identify and characterize the linear epitopes of this allergen, a combination of chemical and enzymatic cleavage and immunoblotting techniques, with subsequent N-terminal sequencing and mass spectrometry, were performed. At least 13 different linear epitopes reacting with the rabbit anti-Asp f 13 antiserum were identified, located throughout the entire molecule. In contrast, IgE from A. fumigatus-sensitive patients bound to three immunodominant epitopes at the C-terminal of the protein. PMID- 10677363 TI - Protein phosphatase 2A is associated in an inactive state with microtubules through 2A1-specific interaction with tubulin. AB - Protein phosphatase (PP) 2A1, a trimer composed of A-, B- and C-subunits in the PP2A family, has been regarded as a principal form localizing at microtubules (MT), but PP2A2, the dimer of A- and C-subunits, has not. Substantiating the claim, the present work shows that the PP2A1 but not PP2A2, both isolated from bovine extract, largely associated with the purified preparation of MT. Furthermore, PP2A1 was found to bind purifiedtubulin polymerized by taxol. The presence of MT associated proteins with purified tubulin hardly affected the binding of PP2A1 to the tubulin. In addition, PP2A1 activity towards glycogen phosphorylase, a probably unphysiological but good substrate, was similarly inhibited by MT proteins and purified tubulin, which accounts for > or =85% of MT proteins, with their IC(50) of about 0.15 mg/ml. In contrast, the inhibition of PP2A2 was about 40% with 1 mg/ml MT proteins and 20% with 0.8 mg/ml tubulin, consistent with its weak association with MT. Therefore, the association with and resultant inhibition by MT proteins of PP2A1 is largely effected by the binding of PP2A1 to tubulin molecule. Moreover, PP2A1 isolated from MT has higher affinity for polymerized MT proteins than has PP2A1 from the postmicrotubule supernatant. The MT PP2A1 has also higher sensitivity to the inhibition by tubulin and MT proteins than has the supernatant PP2A1 (IC(50): 0.1-0.2 mg/ml vs. 0.3-0.6 mg/ml), demonstrating the importance of its association with polymerized tubulin. PMID- 10677364 TI - Identification of a DNA-binding domain and an active-site residue of pseudorabies virus DNase. AB - The pseudorabies virus (PRV) DNase gene has an open reading frame of 1476 nt, capable of coding a 492-residue protein. A previous study showed that PRV DNase is an alkaline exonuclease and endonuclease, exhibiting an Escherichia coli RecBCD-like catalytic function. To analyse its catalytic mechanism further, we constructed a set of clones truncated at the N-terminus or C-terminus of PRV DNase. The deleted mutants were expressed in E. coli with the use of pET expression vectors, then purified to homogeneity. Our results indicate that (1) the region spanning residues 274-492 exhibits a DNA-binding ability 7-fold that of the intact DNase; (2) the N-terminal 62 residues and the C-terminal 39 residues have important roles in 3'-exonuclease activity, and (3) residues 63-453 are responsible for 5'- and 3'-exonuclease activities. Further chemical modification of PRV DNase revealed that the inactivation of DNase by diethyl pyrocarbonate, which was reversible on treatment with hydroxylamine, seemed to be attributable solely to the modification of histidyl residues. Because the herpesviral DNases contained only one well-conserved histidine residue, site directed mutagenesis was performed to replace His(371) with Ala. The mutant lost most of its nuclease activity; however, it still exhibited a wild-type level of DNA-binding ability. In summary, these results indicate that PRV DNase contains an independent DNA-binding domain and that His(371) is the active-site residue that has an essential role in PRV DNase activity. PMID- 10677365 TI - Characterization of peptidyl boronic acid inhibitors of mammalian 20 S and 26 S proteasomes and their inhibition of proteasomes in cultured cells. AB - Proteasomes are large multisubunit proteinases which have several distinct catalytic sites. In this study a series of di- and tri-peptidyl boronic acids have been tested on the chymotrypsin-like activity of purified mammalian 20 S and 26 S proteasomes assayed with succinyl-Leu-Leu-Val-Tyr-amidomethylcoumarin (suc Leu-Leu-Val-Tyr-AMC) as substrate. The inhibition of 20 S proteasomes is competitive but only slowly reversible. The K(i) values for the best inhibitors were in the range 10-100 nM with suc-Leu-Leu-Val-Tyr-AMC as substrate, but the compounds tested were much less effective on other proteasome activities measured with other substrates. Free boronic acid inhibitors exhibited equivalent potency to their pinacol esters. Both benzoyl (Bz)-Phe-boroLeu and benzyloxycarbonyl (Cbz)-Leu-Leu-boroLeu pinacol ester inhibited 20 S and 26 S proteasomes with non ideal behaviour, differences in inhibition of the two forms of proteasomes becoming apparent at high inhibitor concentrations (above 3xK(i)). Both of these compounds were also potent inhibitors of 20 S and 26 S proteasomes in cultured cells. However, gel filtration of cell extracts prepared from cells treated with radiolabelled phenacetyl-Leu-Leu-boroLeu showed that only 20 S proteasomes were strongly labelled, demonstrating differences in the characteristics of inhibition of 20 S and 26 S proteasomes. The usefulness of peptidyl boronic acid inhibitors for investigations of proteasome-mediated protein degradation was confirmed by the observation that Bz-Phe-boroLeu and Cbz-Leu-Leu-boroLeu pinacol ester inhibited NFkappaB activation with IC(50) values comparable to their K(i) values for purified proteasomes. The latter result supports the view that the chymotrypsin-like activity of proteasomes assayed with suc-Leu-Leu-Val-Tyr-AMC is a critical one for protein degradation in cells. PMID- 10677367 TI - Expression of heparan sulphate L-iduronyl 2-O-sulphotransferase in human kidney 293 cells results in increased D-glucuronyl 2-O-sulphation. AB - Functionally important interactions between heparan sulphate and a variety of proteins depend on the precise location of O-sulphate groups. Such residues occur at C-2 of L-iduronic (IdoA) and D-glucuronic acid (GlcA) units, and at C-3 and C 6 of D-glucosamine (GlcN) units. Stable transfection of human embryonic kidney 293 cells with a cDNA encoding mouse mastocytoma IdoA 2-O-sulphotransferase resulted in an approx. 6-fold increase in O-sulphotransferase activity, compared with control cells, as determined using O-desulphated heparin as an acceptor. Structural analysis of endogenous heparan sulphate in the transfected cells, following metabolic labelling with either [(3)H]GlcN or [(35)S]sulphate, showed appreciable formation of -GlcA(2-OSO(3))-GlcNSO(3)- disaccharide units (6% of total disaccharide units; 17% of total O-sulphated disaccharide units) that were essentially absent from heparan sulphate from control cells. The increase in GlcA 2-O-sulphation was accompanied by a decrease in the amount of IdoA formed, whereas overall 2-O-sulphation or 6-O-sulphation remained largely unaffected. These findings indicate that 2-O-sulphation of IdoA and GlcA residues is catalysed by the same enzyme in heparan sulphate biosynthesis. PMID- 10677368 TI - Tandem mass spectrometric analysis of aspergillus niger pectin methylesterase: mode of action on fully methyl-esterified oligogalacturonates. AB - The substrate specificity and the mode of action of Aspergillus niger pectin methylesterase (PME) was determined using both fully methyl-esterified oligogalacturonates with degrees of polymerization (DP) 2-6 and chemically synthesized monomethyl trigalacturonates. The enzymic activity on the different substrates and a preliminary characterization of the reaction products were performed by using high-performance anion-exchange chromatography at neutral pH. Electrospray ionization tandem MS (ESI-MS/MS) was used to localize the methyl esters on the (18)O-labelled reaction products during the course of the enzymic reaction. A. niger PME is able to hydrolyse the methyl esters of fully methyl esterified oligogalacturonates with DP 2, and preferentially hydrolyses the methyl esters located on the internal galacturonate residues, followed by hydrolysis of the methyl esters towards the reducing end. This PME is unable to hydrolyse the methyl ester of the galacturonate moiety at the non-reducing end. PMID- 10677366 TI - Involvement of Gialpha2 in sodium butyrate-induced erythroblastic differentiation of K562 cells. AB - The chronic myelogenous leukaemia cell line K562 can be triggered in culture to differentiate along the erythrocytic pathway in response to a variety of stimulatory agents. In the presence of sodium butyrate, these cells differentiate to erythroblasts and acquire the capability to synthesize haemoglobin. We used this cell system to study alterations in the levels of several G-protein subunits during the cell differentiation programme and to assess the involvement of G(i)alpha2 in this process. Western immunoblot analysis revealed the presence of G(s)alpha1, G(s)alpha2, G(i)alpha2, G(q)alpha, Galpha(12), Gbeta1 and Gbeta2 in K562 cells. G(o)alpha, G(z)alpha, Galpha(13) and Galpha(16) were not detected. Although the levels of several G-protein subunits were altered after treatment with sodium butyrate, the most striking change was the robust increase in the levels of G(i)alpha2, which was accompanied by an increase in the mRNA for G(i)alpha2. Inactivation of G(i)alpha2 by adding Bordetella pertussis toxin to the cultures inhibited erythroblastic differentiation by as much as 62%, as measured by haemoglobin accumulation. Furthermore, the addition of an oligonucleotide anti-sense to G(i)alpha2 inhibited the sodium butyrate-induced robust increase in G(i)alpha2 levels, decreasing it to the basal levels seen in control cells; this treatment decreased the erythroblastic differentiation of the cells (as measured by haemoglobin expression) by 50%. Taken together, these findings imply that increased levels of G(i)alpha2 contribute to the sodium butyrate-induced erythroblastic differentiation of K562 cells. PMID- 10677369 TI - Cloning and functional characterization of the 5'-flanking region of human methionine adenosyltransferase 1A gene. AB - Methionine adenosyltransferase (MAT) is an essential cellular enzyme which catalyses the formation of S-adenosylmethionine, the principal methyl donor and precursor for polyamines. In mammals, two different genes, MAT1A and MAT2A, encode for liver-specific and non-liver-specific MAT respectively. We previously described a switch in the MAT expression from MAT1A to MAT2A in human liver cancer, which offered the cancerous cell a growth advantage. Loss of MAT1A expression was due to lack of gene transcription. To study regulation of the MAT1A gene, we have cloned and characterized a 1.9 kb 5'-flanking region of the human MAT1A gene. One transcriptional start site, located 25 nt downstream from a consensus TATA box, was identified by primer extension and RNase protection assays. The promoter contains several consensus binding sites for CAAT enhancer binding protein (C/EBP) and hepatocyte-enriched nuclear factor (HNF), transcriptional factors important in liver-specific gene expression. The human MAT1A promoter was able to efficiently drive luciferase expression in Chang cells, a human liver cell line, but not in HeLa cells. Sequential deletion analysis of the promoter revealed two DNA regions upstream of the translational start site, -705 to -839 bp and -1111 to -1483 bp, which are involved in positive and negative gene regulation, respectively. Specific protein binding to these regions was confirmed by electrophoretic-mobility-shift and DNase I footprinting assays. Similar to the situation with the rat MAT1A, glucocorticoid treatment also increased human MAT1A expression and promoter activity in a dose- and time dependent manner. PMID- 10677370 TI - The N-terminal 34 residues of the 55 kDa regulatory subunits of phosphoinositide 3-kinase interact with tubulin. AB - There are five regulatory subunit isoforms of phosphoinositide 3-kinase (PI 3 kinase), which are classified into three groups: proteins of 85 kDa (p85alpha and p85beta), 55 kDa (p55alpha and p55gamma) and 50 kDa (p50alpha). Structural differences between the three groups reside in the N-terminus. To elucidate the unique functional role of the 55 kDa regulatory subunits, GST (glutathione S transferase) fusion proteins containing a unique N-terminal portion consisting of a 34-amino-acid sequence of p55alpha or p55gamma (GST-p55alpha/gammaN(1-34)) were used as affinity matrices to screen rat brain cell extracts for proteins to which this portion binds specifically. A protein that bound was identified as beta tubulin by protein sequencing. In addition, not only the beta isoform of tubulin, but also the alpha and gamma isoforms, were detected in the protein absorbed from cell lysates with GST-p55gammaN(1-34) and GST-p55alphaN(1-34) by immunoblotting. Indeed, the only regulatory subunit present in the purified microtubule assembly from rat brain was the 55 kDa isoform; neither 85 kDa nor 50 kDa subunits were detected. These results indicate endogenous binding of 55 kDa regulatory subunits of PI 3-kinase to tubulin in the brain. Finally, we measured tubulin-associated PI 3-kinase activity in CHO/IR cells overexpressing each of the five regulatory subunit isoforms. Only in cells expressing p55alpha or p55gamma was there a significant elevation of tubulin-associated PI 3-kinase activity in response to insulin. These results suggest that the p55alpha and p55gamma regulatory subunits have important roles in regulating PI 3-kinase activity, particularly for microtubules at the cell periphery. PMID- 10677371 TI - Vitamin C protects against and reverses specific hypochlorous acid- and chloramine-dependent modifications of low-density lipoprotein. AB - Activated phagocytes produce the highly reactive oxidant hypochlorous acid (HOCl) via the myeloperoxidase-catalysed reaction of hydrogen peroxide with chloride ions. HOCl reacts readily with a number of susceptible targets on apolipoprotein B-100 of low-density lipoprotein (LDL), resulting in uncontrolled uptake of HOCl modified LDL by macrophages. We have investigated the effects of vitamin C (ascorbate), an effective water-soluble antioxidant, on the HOCl- and chloramine dependent modification of LDL. Co-incubation of vitamin C (25-200 microM) with LDL resulted in concentration-dependent protection against HOCl (25-200 microM) mediated oxidation of tryptophan and lysine residues, formation of chloramines and increases in the relative electrophoretic mobility of LDL. Vitamin C also partially protected against oxidation of cysteine residues by HOCl, and fully protected against oxidation of these residues by the low-molecular-mass chloramines, N(alpha)-acetyl-lysine chloramine and taurine chloramine, and to a lesser extent monochloramine (each at 25-200 microM). Further, we found that HOCl (25-200 microM)-dependent formation of chloramines on apolipoprotein B-100 was fully reversed by 200 microM vitamin C; however, the loss of lysine residues and increase in relative electrophoretic mobility of LDL were only partially reversed, and the loss of tryptophan and cysteine residues was not reversed. Time course experiments showed that the reversal by vitamin C of HOCl-dependent modifications became less efficient as the LDL was incubated for up to 4 h at 37 degrees C. These data show that vitamin C not only protects against, but also reverses, specific HOCl- and chloramine-dependent modifications of LDL. As HOCl mediated LDL modifications have been strongly implicated in the pathogenesis of atherosclerosis, our data indicate that vitamin C could contribute to the anti atherogenic defence against HOCl. PMID- 10677372 TI - GTPase mechanism and function: new insights from systematic mutational analysis of the phosphate-binding loop residue Ala30 of Rab5. AB - Structural and biochemical data indicate the importance of the phosphate-binding loop residues Gly(12) and Gly(13) of Ras both in the GTP hydrolysis reaction and in biological activity, but these two residues are not conserved in other Ras related GTPases. To gain a better understanding of this region in GTP hydrolysis and GTPase function, we used the Ras-related Rab5 GTPase as a model for comparison, and substituted the Ala(30) residue (the equivalent of Gly(13) of Ras) with all the other 19 amino acids. The resulting mutants were analysed for GTP hydrolysis, GTP binding, GTP dissociation and biological activity. Only the substitution of alanine with proline reduced the GTPase activity by an order of magnitude. This effect is in sharp contrast with the observation that a proline substitution at the neighbouring position (Gly(12) of Ras) has little effect on the GTPase activity. Whereas most other substitutions showed either a small negative effect or no effect on the GTPase activity, the arginine substitution surprisingly stimulated the GTPase activity by 5-fold. Molecular modelling suggests that this built-in arginine mimics the catalytic arginine residues found in trimeric GTPases and GTPase-activating proteins in providing the positive charge to facilitate the GTP hydrolysis reaction. We investigated further the biological activity of the Rab5 mutants in relation to stimulating endocytosis. When expressed in cultured baby hamster kidney cells, both arginine and proline mutants, like wild-type Rab5, stimulated endocytosis. However, the arginine mutant was a more potent stimulator than the proline mutant (3-fold stimulation as against 1.7-fold). The tryptophan mutant, on the other hand, was completely deficient in activity in terms of the stimulation of endocytosis, demonstrating the importance of the phosphate-binding loop in Rab GTPase function. PMID- 10677373 TI - Mutations in the reduced-folate carrier affect protein localization and stability. AB - The reduced-folate-carrier (rfc) gene has been shown to be functionally important for reduced-folate transport in mammalian cells. In the present paper we describe the identification of alterations in both alleles of the rfc gene in a mutant Chinese-hamster ovary cell line deficient in methotrexate transport. One allele of the rfc gene contains a point mutation resulting in a Gly(345)-->Arg substitution in the predicted amino acid sequence. In this case, a protein of similar size to the wild-type protein is produced, although it remains as an immature, core-glycosylated, form. The second allele contains a point mutation in the last base of intron 5 that results in the utilization of a cryptic splice site leading to a seven-base deletion in the mRNA. The use of an alternate splice site changes the reading frame to yield a truncated protein with 68 different C terminal amino acids as compared with the wild-type. Both of these altered gene products were monitored by fusion with green fluorescent protein and found to be non-functional with an increased rate of turnover. The protein with the point mutation is trapped in the endoplasmic reticulum with subsequent degradation, whereas the product of the splice mutation is not membrane-associated and is partially degraded. Thus mutations in both alleles of the rfc gene in this resistant cell line account for the loss of reduced-folate transport. The observations made regarding the degradation of these mutant gene products also provide support for putative checkpoints in the endoplasmic reticulum. PMID- 10677374 TI - Crystal structure of the anti-(carcinoembryonic antigen) single-chain Fv antibody MFE-23 and a model for antigen binding based on intermolecular contacts. AB - MFE-23 is the first single-chain Fv antibody molecule to be used in patients and is used to target colorectal cancer through its high affinity for carcinoembryonic antigen (CEA), a cell-surface member of the immunoglobulin superfamily. MFE-23 contains an N-terminal variable heavy-chain domain joined by a (Gly(4)Ser)(3) linker to a variable light-chain (V(L)) domain (kappa chain) with an 11-residue C-terminal Myc-tag. Its crystal structure was determined at 2.4 A resolution by molecular replacement with an R(cryst) of 19.0%. Five of the six antigen-binding loops, L1, L2, L3, H1 and H2, conformed to known canonical structures. The sixth loop, H3, displayed a unique structure, with a beta-hairpin loop and a bifurcated apex characterized by a buried Thr residue. In the crystal lattice, two MFE-23 molecules were associated back-to-back in a manner not seen before. The antigen-binding site displayed a large acidic region located mainly within the H2 loop and a large hydrophobic region within the H3 loop. Even though this structure is unliganded within the crystal, there is an unusually large region of contact between the H1, H2 and H3 loops and the beta-sheet of the V(L) domain of an adjacent molecule (strands DEBA) as a result of intermolecular packing. These interactions exhibited remarkably high surface and electrostatic complementarity. Of seven MFE-23 residues predicted to make contact with antigen, five participated in these lattice contacts, and this model for antigen binding is consistent with previously reported site-specific mutagenesis of MFE-23 and its effect on CEA binding. PMID- 10677375 TI - Interaction between the homeodomain proteins Cdx2 and HNF1alpha mediates expression of the lactase-phlorizin hydrolase gene. AB - Lactase-phlorizin hydrolase is a brush-border enzyme which is specifically expressed in the small intestine where it hydrolyses lactose, the main carbohydrate found in milk. We have previously demonstrated in transgenic mice that the tissue-specific and developmental expression of lactase is controlled by a 1 kb upstream region of the pig lactase gene. Two homeodomain transcription factors, caudal-related homeodomain protein (Cdx2) and hepatic nuclear factor 1alpha (HNF1alpha), are known to bind to regulatory cis elements in the promoters for several intestine-specific genes, including lactase, and are present in mammalian intestinal epithelia from an early stage in development. In the present study, we examined whether Cdx2 and HNF1alpha physically interact and co operatively activate transcription from the lactase-phlorizin hydrolase promoter. We show that the presence of both factors leads to a much higher level of transcription than the sum of the activation by either factor alone. The N terminal activation domain of Cdx2 is required for maximal synergy with HNF1alpha. With the use of pull-down assays, we demonstrate a direct protein protein interaction between Cdx2 and HNF1alpha. The interaction domain includes the homeodomain region of both proteins. This is the first demonstration of a functional interaction between two transcription factors involved in the activation of a number of intestine-specific genes. Synergistic interaction between tissue-restricted factors is likely to be an important mechanism for reinforcing developmental and tissue-specific gene expression within the intestine. PMID- 10677376 TI - Rapid replenishment of sphingomyelin in the plasma membrane upon degradation by sphingomyelinase in NIH3T3 cells overexpressing the phosphatidylinositol transfer protein beta. AB - In order to study the in vivo function of the phosphatidylinositol transfer protein beta (PI-TPbeta), mouse NIH3T3 fibroblasts were transfected with cDNA encoding mouse PI-TPbeta. Two stable cell lines were isolated (SPIbeta2 and SPIbeta8) in which the levels of PI-TPbeta were increased 16- and 11-fold respectively. The doubling time of the SPIbeta cells was about 1.7 times that of the wild-type (wt) cells. Because PI-TPbeta expresses transfer activity towards sphingomyelin (SM) in vitro, the SM metabolism of the overexpressors was investigated. By measuring the incorporation of [methyl-(3)H]choline chloride in SM and phosphatidylcholine (PtdCho), it was shown that the rate of de novo SM and PtdCho synthesis was similar in transfected and wt cells. We also determined the ability of the cells to resynthesize SM from ceramide produced in the plasma membrane by the action of bacterial sphingomyelinase (bSMase). In these experiments the cells were labelled to equilibrium (60 h) with [(3)H]choline. At relatively low bSMase concentrations (50 munits/ml), 50% of [(3)H]SM in wt NIH3T3 cells was degraded, whereas the levels of [(3)H]SM in SPIbeta cells appeared to be unaffected. Since the release of [(3)H]choline phosphate into the medium was comparable for both wt NIH3T3 and SPIbeta cells, these results strongly suggest that breakdown of SM in SPIbeta cells was masked by rapid resynthesis of SM from the ceramide formed. By increasing the bSMase concentrations to 200 munits/ml, a 50% decrease in the level of [(3)H]SM in SPIbeta cells was attained. During a recovery period of 6 h (in the absence of bSMase) the resynthesis of SM was found to be much more pronounced in these SPIbeta cells than in 50% [(3)H]SM-depleted wt NIH3T3 cells. After 6 h of recovery about 50% of the resynthesized SM in the SPIbeta cells was available for a second hydrolysis by bSMase. When monensin was present during the recovery period, the resynthesis of SM in bSMase-treated SPIbeta cells was not affected. However, under these conditions 100% of the resynthesized SM was available for hydrolysis. On the basis of these results we propose that, under conditions where ceramide is formed in the plasma membrane, PI-TPbeta plays an important role in restoring the steady-state levels of SM. PMID- 10677377 TI - Plasmodium falciparum-infected red blood cells depend on a functional glutathione de novo synthesis attributable to an enhanced loss of glutathione. AB - During the erythrocytic cycle, Plasmodium falciparum is highly dependent on an adequate thiol status for its survival. Glutathione reductase as well as de novo synthesis of GSH are responsible for the maintenance of the intracellular GSH level. The first and rate-limiting step of the synthetic pathway is catalysed by gamma-glutamylcysteine synthetase (gamma-GCS). Using L-buthionine-(S, R) sulphoximine (BSO), a specific inhibitor of the gamma-GCS, we show that the infection with P. falciparum causes drastic changes in the GSH metabolism of red blood cells (RBCs). Infected RBCs lose GSH at a rate 40-fold higher than non infected RBCs. The de novo synthesis of the tripeptide was found to be essential for parasite survival. GSH depletion by BSO inhibits the development of P. falciparum with an IC(50) of 73 microM. The effect of the drug is abolished by supplementation with GSH or GSH monoethyl ester. Our studies demonstrate that the plasmodicidal effect of the inhibitor BSO does not depend on its specificity towards its target enzyme in the parasite, but on the changed physiological needs for the metabolite GSH in the P. falciparum-infected RBCs. Therefore the depletion of GSH is proposed as a chemotherapeutic strategy for malaria, and gamma-GCS is proposed as a potential drug target. PMID- 10677378 TI - Modulation of the glutathione S-transferase in Ochrobactrum anthropi: function of xenobiotic substrates and other forms of stress. AB - The gluthathione S-transferase gene of the atrazine-degrading bacterium Ochrobactrum anthropi (OaGST) encodes a single-subunit polypeptide of 201 amino acid residues (Favaloro et al. 1998, Biochem. J. 335, 573-579). RNA blot analysis showed that the gene is transcribed into an mRNA of about 800 nucleotides, indicating a monocistronic transcription of the OaGST gene. The modulation of OaGST in this bacterium, in the presence of different stimulants, was investigated. The level of expression of OaGST was detected both by measuring the mRNA level and by immunoblotting experiments. OaGST is a constitutive enzyme which is also inducible by several stimulants. In fact, atrazine caused an increase in the expression of OaGST even at concentrations which had no effect on growth rates of the bacteria. Moreover, the presence of other aromatic substrates of this bacterium, such as phenol and chlorophenols, leads to a marked enhancement in OaGST expression. In this case, the expression of OaGST was related to growth inhibition and membrane damage caused by these hydrophobic compounds, and to the adaptive responses of the cell membranes. On the other hand, toluene and xylene, two aromatic compounds not degradable by this bacterium, did not induce the OaGST expression. The same was observed for other stress conditions such as low pH, heat shock, hydrogen peroxide, osmotic stress, starvation, the presence of aliphatic alcohols or heavy metals. These results suggest a co-regulation of the OaGST gene by the catabolic pathways of phenols and chlorophenols in this bacterium. Therefore, OaGST could function as a detoxifying agent within the catabolism of these xenobiotics. PMID- 10677379 TI - Autonomic function in normal pregnancy: the role of studying heart rate variability. PMID- 10677380 TI - Nitric oxide and the airway. PMID- 10677381 TI - Identifying genetic susceptibility factors for tuberculosis in Africans: a combined approach using a candidate gene study and a genome-wide screen. AB - There is convincing evidence that host genes affect the outcome of infection in human tuberculosis. Two complementary strategies were used to identify the genes involved. A linkage-based genome-wide screen was carried out to locate the positions of genes exerting a major population-wide effect on tuberculosis susceptibility. A candidate-gene-based case-control study was used to examine the effects of genes that may exert a more moderate effect on risk of clinical tuberculosis. The genome screen was conducted in two stages. In the first stage 299 microsatellite markers, spanning all 23 chromosomes, were typed in 92 independent sib-pairs, and seven regions showed some evidence of co-segregation with the disease. These seven regions were examined in a second set of 81 sib pairs, and markers on chromosomes 15q and Xq showed evidence of linkage to tuberculosis. An X chromosome susceptibility gene may contribute to the excess of males with tuberculosis observed in many populations. The candidate gene approach compared the frequency of polymorphisms in several genes in over 400 subjects with smear-positive pulmonary tuberculosis and 400 ethnically matched healthy controls. Polymorphisms in genes encoding natural-resistance-associated macrophage protein, vitamin D receptor and mannose-binding lectin were associated with tuberculosis. These results suggest that many genes may be involved in determining host susceptibility to tuberculosis, and highlight the importance of using several different study methods to locate them. PMID- 10677382 TI - Genetic analysis of the atrial natriuretic peptide gene in essential hypertension. AB - Atrial natriuretic peptide (ANP) plays an important role in the regulation of blood pressure through sodium-water homoeostasis. Accordingly, several investigators have raised the question of whether the gene encoding ANP is involved in the aetiology of essential hypertension or related phenotypes such as salt sensitivity. Most of the studies have used anonymous polymorphic markers of the gene, and made inconclusive claims about the disease relevance of ANP. Therefore, in order to find sequence variations with potential functional significance and to characterize the pattern of linkage disequilibrium between polymorphisms, we screened a 3368-bp genomic fragment of ANP. Subsequently we tested the association of detected polymorphisms with plasma ANP levels and with hypertension status. Two new polymorphisms were identified, in the 5' untranslated region and exon 1 respectively, as well as three previously reported polymorphisms in intron 2 and exon 3. When analysed in 102 healthy normotensive subjects, none of the polymorphisms appeared to significantly affect plasma ANP levels. A case-control study in a Japanese population (255 hypertensive and 225 normotensive individuals) revealed a marginally significant association (P=0.026) between an ANP polymorphism located in the 5'-untranslated region (C-664G) and hypertension, but no association for the other polymorphisms. Each of the uncommon variants has an allele frequency of less than 10% in Japanese people, which may have hampered our detection of a significant association between ANP variants and hypertension status (and plasma ANP levels). The pathophysiological relevance of ANP, however, needs to be further defined in relation to hypertension-associated phenotypes, and also should be examined in different ethnic groups. PMID- 10677383 TI - Changes in baroreceptor sensitivity for heart rate during normotensive pregnancy and the puerperium. AB - Normal pregnancy is associated with marked changes in cardiovascular haemodynamics, which in part may be due to changes in autonomic control mechanisms. Baroreflex sensitivity for heart rate (BRS) was calculated in the supine and standing positions using power spectral analysis of pulse interval (PI) and systolic blood pressure (SBP) in 16 normotensive pregnant women and 10 normotensive non-pregnant controls. The pregnant women were studied on three occasions during their pregnancy (early, mid- and late gestation) and once during the puerperium. Supine total SBP variability increased between early and late pregnancy by 79% [95% confidence intervals (CI) 30%, 145%; P<0. 001], and supine high-frequency PI variability decreased by 75% (CI 51%, 88%; P<0.001). Supine BRS fell by 50% (P<0.001), with values returning to early-pregnancy levels in the puerperium, which were similar to those recorded in the control group. Standing SBP variability and BRS values were unchanged during pregnancy and post partum. The low/high frequency ratio of PI variability, taken as a surrogate measure of sympathovagal balance, increased by 137% (CI 42%, 296%; P<0.01) in the supine but not the standing position from early to late pregnancy. This was due to a decrease in high-frequency variability rather than to an increase in low frequency variability, suggesting that these changes may have been due to vagal withdrawal rather than increased sympathetic activity. Normotensive pregnancy is associated with a marked decrease in supine BRS, although the exact mechanisms for these changes remain unclear. Further studies are required to define whether changes in BRS and sympathovagal tone in early pregnancy can be used to predict the onset of pregnancy-induced hypertension. PMID- 10677384 TI - Antihypertensive treatment in early postnatal life modulates prenatal dietary influences upon blood pressure in the rat. AB - Epidemiological evidence from diverse human populations, supported by experimental evidence from animal models, suggests that maternal nutrition during pregnancy is an important fetal programming influence upon cardiovascular disease. Experiments with a low-protein-diet model of rat pregnancy suggest a role for the renin-angiotensin system in the programming mechanism, since fetal undernutrition permanently elevates pulmonary and plasma angiotensin-converting enzyme activity. Long-term beneficial effects of captopril on blood pressure in this model require further investigation in order to clarify the role of angiotensin II. Pregnant rats were fed a control diet containing 18% (w/w) casein as the protein source or a low-protein diet containing 9% (w/w) casein. Between the ages of 2 and 4 weeks postnatally, mothers and their pups were treated with losartan or nifedipine. All pups in the study had blood pressure determined at 4 and 12 weeks of age using a tail cuff. Animals exposed to the low-protein diet in utero and not subjected to drug treatment had elevated blood pressure relative to control rats (mean increase of 27 mmHg; P<0.001). Treatment of rats exposed to the control diet in utero with either nifedipine or losartan between 2 and 4 weeks of age did not alter their blood pressure. Nifedipine had no effect upon the blood pressure of low-protein-exposed pups, but losartan prevented the blood pressure elevation in these animals. Between 4 and 12 weeks of age, blood pressure increased significantly in all groups (P<0.001). The pattern of blood pressure among the groups was identical to that observed at 4 weeks, suggesting that the observed early effects of losartan would be maintained into adult life. The data are consistent with the hypothesis that angiotensin II plays a major role in the prenatal programming of hypertension. The action of angiotensin II at the AT(1) receptor between 2 and 4 weeks of age may be critically up-regulated by fetal factors, including exposure to glucocorticoids of maternal origin. PMID- 10677385 TI - Cytochrome P450 metabolites of arachidonic acid may be important mediators in angiotensin II-induced vasoconstriction in the rat mesentery in vivo. AB - We have investigated the role of cytochrome P450 (CYP-450) metabolites of arachidonic acid in the modulation of vascular reactivity to angiotensin II in vivo using an in situ blood-perfused mesenteric preparation in anaesthetized spontaneously hypertensive rats (SHR). Miconazole, a non-selective inhibitor of CYP-450 that inhibits both hydroxylation and epoxidation, substantially suppressed mesenteric vasoconstrictor responses to angiotensin II in SHR, but had no effect on responses to noradrenaline or sympathetic nerve stimulation. In normotensive Wistar-Kyoto (WKY) rats, miconazole caused only a modest suppression of vasoconstrictor responses to angiotensin II. N-Methylsulphonyl-12, 12 dibromododec-11-enamide (DDMS), a new selective inhibitor of CYP-450 omega hydroxylase activity, decreased mean intra-arterial blood pressure and significantly attenuated mesenteric angiotensin II-induced vasoconstrictor responses in SHR. Isolated mesenteric vessels were able to metabolize (14)C labelled arachidonic acid to hydroxyeicosatetraenoic acids (HETEs) in vitro, and this was substantially inhibited by DDMS. The results from the present studies combined with the existing evidence that angiotensin II stimulates the release of 20-HETE, a CYP-450 metabolite of arachidonic acid, suggest that CYP-450-derived HETEs may be important mediators in angiotensin II-induced vasoconstriction. However, the development of more sensitive assays for the detection in vivo of 20 HETE in mesenteric vessels would be required to confirm these findings. PMID- 10677386 TI - Effects of proinsulin C-peptide on nitric oxide, microvascular blood flow and erythrocyte Na+,K+-ATPase activity in diabetes mellitus type I. AB - This study was conducted to evaluate the influence of proinsulin C-peptide on erythrocyte Na(+),K(+)-ATPase and endothelial nitric oxide synthase activities in patients with type I diabetes. In a randomized double-blind study design, ten patients with type I diabetes received intravenous infusions of either human C peptide or physiological saline on two different occasions. C-peptide was infused at a rate of 3 pmol.min(-1).kg(-1) for 60 min, and thereafter at 10 pmol.min( 1).kg(-1) for 60 min. At baseline and after 60 and 120 min, laser Doppler flow (LDF) was measured following acetylcholine iontophoresis or mild thermal stimulation (44 degrees C), and venous blood samples were collected to determine plasma cGMP levels and erythrocyte membrane Na(+),K(+)-ATPase activity. The LDF response to acetylcholine increased during C-peptide infusion and decreased during saline infusion [18.6+/-19.2 and -13.2+/-9.4 arbitrary units respectively; mean+/-S.E.M.; P<0.05). No significant change in LDF was observed after thermal stimulation. The baseline plasma concentration of cGMP was 5.5+/-0.6 nmol.l(-1); this rose to 6.8+/-0.9 nmol.l(-1) during C-peptide infusion (P<0.05). Erythrocyte Na(+),K(+)-ATPase activity increased from 140+/-29 nmol of P(i).h(-1).mg(-1) in the basal state to 287+/-5 nmol of P(i). h(-1).mg(-1) during C-peptide infusion (P<0.01). There was a significant linear relationship between plasma C-peptide levels and erythrocyte Na(+),K(+)-ATPase activity during the C-peptide infusion (r=0.46, P<0.01). No significant changes in plasma cGMP levels or Na(+),K(+) ATPase activity were observed during saline infusion. This study demonstrates an effect of human proinsulin C-peptide on microvascular function, which might be mediated by an increase in NO production and an activation of the erythrocyte Na(+),K(+)-ATPase. These mechanisms are compatible with the previous observed microvascular effects of C-peptide in patients with type I diabetes. PMID- 10677387 TI - Role of nitric oxide in airway remodelling. AB - Airway remodelling, which is manifested by thickening of bronchial wall, is an important causative factor of bronchial hyper-responsiveness in asthma. The pathophysiological mechanism of airway remodelling is not clear. In the present study we evaluated the relationship between nitric oxide (NO) generation and airway wall thickening in patients with chronic asthma. As a marker of NO production, the levels of nitrite/nitrate were measured in induced sputum, and bronchial wall thickening was measured by high-resolution computed tomography. Sputum concentrations of nitrite/nitrate were significantly increased in asthmatic patients compared with controls. The ratio of airway wall thickness to lumen diameter was significantly correlated with the sputum concentration of nitrite/nitrate. Although statistical correlation does not prove causation, this finding suggests that NO may play a key role in the pathogenesis of airway remodelling. PMID- 10677388 TI - Albumin stimulates p44/p42 extracellular-signal-regulated mitogen-activated protein kinase in opossum kidney proximal tubular cells. AB - The presence of protein in the urine of patients with renal disease is an adverse prognostic feature. It has therefore been suggested that proteinuria per se may be responsible for the development of renal tubulo-interstitial scarring and fibrosis, and disturbances in tubular cell growth and proliferation. We have used the opossum kidney proximal tubular cell line to investigate the effects of albumin on cell growth. The effect of albumin on cell proliferation was investigated by cell counting and measurement of [(3)H]thymidine incorporation. We studied the effect of recombinant human albumin on the activity of p44/p42 extracellular-signal-regulated mitogen-activated protein kinase (MAP kinase ) using an in vitro kinase assay, and immunoblotting with antibodies against active extracellular-signal-regulated kinase (ERK). The effects of the ERK inhibitor PD98059 were also examined. Recombinant human albumin was found to stimulate proliferation of opossum kidney cells in a dose-dependent manner, with maximal stimulation at a concentration of 1 mg/ml. In addition, recombinant human albumin activated ERK in a time-dependent (maximal after 5 min) and dose-dependent (maximal at 1 mg/ml) fashion. These effects on cell proliferation and ERK activity were inhibited by PD98059, and were not reproduced by ovalbumin or mannitol. The data therefore indicate that albumin is able to stimulate growth and proliferation of proximal tubular cells that is dependent on the ERK family of MAP kinases. The potential importance of this pathway in the development of renal disease is discussed. PMID- 10677389 TI - Effects of adenosine receptor antagonists on the responses to contrast media in the isolated rat kidney. AB - Contrast media can induce both a decrease in renal blood flow and a reduction in glomerular filtration rate (GFR) when administered to both experimental animals and humans. In the present study we have examined the role of adenosine in mediating these effects using the isolated perfused rat kidney. Kidneys were perfused with a 6. 7%-(w/v)-albumin-based perfusate supplemented with glucose and amino acids (n=6 per group). They were exposed to diatrizoate [20 mg of iodine (mgI)/ml; osmolality 1650 mOsm/kg of water] or iotrolan (20 mgI/ml; osmolality 320 mOsm/kg of water) in the presence or absence of theophylline (10.8 microg/ml), or to diatrizoate in the presence or absence of a specific adenosine A(1) receptor antagonist (KW-3902; 2 microg/ml) or a specific A(2) receptor antagonist (KF17837; 6 microg/ml). Diatrizoate (n=6) produced a fall in GFR from 0.65+/-0.04 to 0.42+/-0.03 ml.min(-1).g(-1) (P<0.05); renal perfusate flow (RPF) also declined, from 36.5+/-3.8 to 22.0+/-3.2 ml. min(-1).g(-1) (P<0.05). Iotrolan (n=6) produced a fall in GFR from 0. 64+/-0.02 to 0.48+/-0.04 ml.min(-1).g(-1) (P<0.05) and in RPF from 33.3+/-3.8 to 24.0+/-3.0 ml.min(-1).g(-1) (P<0.05). Theophylline (10.8 microg/ml) prevented the fall in GFR caused by either diatrizoate (baseline, 0.63+/-0.05 ml.min(-1).g(-1); diatrizoate+theophylline, 0. 60+/-0.04 ml.min(-1).g(-1)) or iotrolan (baseline, 0.64+/-0.04 ml. min(-1).g(-1); iotrolan+theophylline, 0.67+/-0.05 ml.min(-1).g(-1)), but did not affect the decreases in RPF caused by either agent. KW-3902 (2 microg/ml) also prevented the fall in GFR produced by diatrizoate (baseline, 0.66+/-0.05 ml.min(-1).g(-1); diatrizoate+KW-3902, 0.61+/-0.05 ml.min(-1).g(-1)), while the fall in RPF remained unaffected. KF17837 (6 microg/ml) had no effect on the decreases in either GFR or RPF induced by diatrizoate (n=6 per group). The results suggest a role for adenosine acting at the A(1) receptor in mediating the decrease in GFR induced by contrast media. This effect is independent of a change in renal vascular resistance, and possibly secondary to mesangial cell contraction causing a decrease in the ultrafiltration coefficient. PMID- 10677390 TI - Does a high concentration of calcium in the urine cause an important renal concentrating defect in human subjects? AB - The objective of this study was to evaluate the hypothesis that a high concentration of ionized calcium in the lumen of the medullary collecting duct causes an osmole-free water diuresis. The urine flow rate and osmolality were measured in normal human subjects, as well as in patients with a history of nephrolithiasis who excreted more than 5 mmol of calcium per 24 h. There was an inverse relationship between the concentration of calcium in the urine and the 24 h urine volume both in normal subjects and in patients with a history of nephrolithiasis. When the concentration of calcium in the urine was greater than 5 mmol/l, the urine volume was less than 1 litre per day in the majority of subjects. After 16 h of water deprivation, when the concentration of calcium in the urine was as high as 17 mmol/l (ionized calcium 7.4 mmol/l), urine osmolality was 1258 mOsm/kg of water and the urine flow rate was 0.30 ml/min. We conclude that, although a calcium receptor may be present in the lumen of the medullary collecting duct in human subjects, an extremely high concentration of urinary total and ionized calcium does not cause a clinically important defect in the renal concentrating process. PMID- 10677391 TI - Myocardial tissue oxygen supply and utilization during coronary artery bypass surgery: evidence of microvascular no-reflow. AB - The supply and utilization of oxygen by the myocardium reflect the dynamic efficiency of the microcirculation. The present study examines these parameters during coronary artery bypass surgery. We used a voltammetric microelectrode technique to assess regional variations in myocardial tissue partial pressure of oxygen (PO(2)) and myocardial tissue perfusion (MTP) in patients undergoing coronary artery bypass surgery. A total of 29 myocardial regions were studied in 17 patients to assay tissue PO(2), and 13 regions in 10 patients to measure MTP. There was an increase in MTP from 53+/-9 ml.min(-1).100 g(-1) before cardiopulmonary bypass to 72+/-13 ml. min(-1).100 g(-1) after (means+/-S.E.M.; P=0.05). Tissue PO(2) showed an overall increase from a baseline level of 45+/-8 mmHg to a final level of 88+/-10 mmHg (P<0.0001). Following release of the aortic cross-clamp there was a variable time delay before a change in tissue PO(2) was observed. There was an immediate response in five regions, whereas in 20 regions the response was delayed by between 0.5 and 32 min. In the remaining four regions there was no change in tissue PO(2). The duration of the delay in response was correlated positively with the cross-clamp time (r=0.45, P<0.05) and negatively with the final tissue PO(2) (r=-0.5, P<0.05). Voltammetric methods for monitoring changes in oxygen supply and utilization offer new insights into the changes that occur during ischaemia and reperfusion. A delay in the delivery of oxygen to the myocardium occurs in many patients following coronary artery bypass surgery. PMID- 10677392 TI - Effects of prolonged hypobaric hypoxia on human skeletal muscle function and electromyographic events. AB - This study tested the hypothesis that a prolonged decrease in arterial oxygen pressure in resting or contracting skeletal muscles alters their ability to develop force through an impairment of energy-dependent metabolic processes and also through an alteration of electrophysiological events. The experiment was conducted during a 32-day simulated ascent of Mt. Everest (8848 m altitude) (Everest III Comex '97), which also allowed testing of the effects of re oxygenation on muscle function. Maximal voluntary contractions (MVCs) of the flexor digitorum, and static handgrips sustained at 60% of MVC, were performed by eight subjects before the ascent (control), then during the stays at simulated altitudes of 5000 m, 6000 m and 7000 m, and finally 1 day after the return to 0 m. The evoked muscle compound action potential (M-wave) was recorded at rest and during the manoeuvres at 60% of MVC. The changes in median frequency of electromyographic (EMG) power spectra were also studied during the contraction at 60% of MVC. In four individuals, transient re-oxygenation during the ascent allowed us to test the reversibility of hypoxia-induced MVC and M-wave changes. At rest, a significant decrease in M-wave amplitude was noted at 5000 m. This effect was associated with a prolonged M-wave conduction time at 6000 m and an increased M-wave duration at 7000 m, and persisted after the return to 0 m. Re oxygenation did not modify the changes in M-wave characteristics. A significant decrease in MVC was measured only during the ascent (-10 to -24%) in the non dominant forearm of subjects who underwent re-oxygenation; this intervention slightly improved muscle strength at 6000 m and 7000 m. During the ascent and after the return to 0 m, there was a significant reduction of the median frequency decrease throughout contraction at 60% of MVC compared with the EMG changes measured before the ascent. It is concluded that prolonged exposure to hypoxia slows the propagation of myopotentials and alters sensorimotor control during sustained effort. Re-oxygenation did not affect the hypoxia-induced EMG changes and had a modest influence on muscle strength. PMID- 10677393 TI - Deleterious effects of chronic clenbuterol treatment on endurance and sprint exercise performance in rats. AB - The beta(2)-adrenergic agonist, clenbuterol, has powerful muscle anabolic and lipolytic effects and is used by athletes to improve exercise performance; however, its use in conjunction with different forms of exercise training has received limited attention. Since previous studies have reported that chronic use of other beta(2)-adrenergic agonists has deleterious effects on cardiac muscle structure and function, the aim of the present study was to determine whether chronic clenbuterol administration would reduce the exercise capabilities of rats subjected to long-term treadmill sprint running, endurance swimming or voluntary wheel running training. The effect of clenbuterol treatment on exercise performance in rats was evaluated in three separate studies. Different groups of male rats were assigned to an endurance swimming (2 h/day, 5/7 days, 18 weeks) group, a treadmill sprint running (8x1 min bouts, 1.05 m/s, 20 weeks) group, or a voluntary wheel running (16 weeks) group. In each study, rats were allocated into either a treated group that received clenbuterol (2 mg.kg(-1).day(-1)) in their drinking water or an untreated control group. In each of the three studies, treated rats exhibited a reduction in exercise performance compared with untreated rats. Treated rats ran approximately 57% less total distance than untreated rats in the voluntary running programme and were unable to complete the swimming and sprinting protocols performed by the untreated rats. In each of the studies, the treated rats exhibited cardiac hypertrophy, with absolute heart mass increased by approximately 19% and heart mass relative to body mass increased by approximately 20%. The hearts of sedentary rats treated with clenbuterol exhibited extensive collagen infiltration surrounding blood vessels and in the wall of the left ventricle. The results indicate strongly that chronic clenbuterol administration deleteriously affects exercise performance in rats, potentially due to alterations in cardiac muscle structure and function. PMID- 10677394 TI - Effects of pre- and post-absorptive factors on the lactulose/rhamnose gut permeability test. AB - It is assumed that the outcome of the lactulose/rhamnose gut permeability test is not influenced by pre- or post-absorptive factors. The aim of our study was to investigate the role of a pre-absorptive factor, i.e. small-intestinal transit, and a post-absorptive factor, i.e. renal clearance. Ten healthy male subjects were studied. Urinary lactulose and rhamnose excretion was measured after intraduodenal administration of lactulose and rhamnose following induction of increased intestinal permeability using chenodeoxycholic acid (chenodiol), in the absence and in the presence of accelerated intestinal transit. Urinary sugar excretion was measured after intravenous administration of either a regular dose (50 mg/50 mg) or a high dose (250 mg/250 mg) of lactulose/rhamnose. The intraduodenal experiments showed that a combination of accelerated small-bowel transit and increased permeability did not lead to significant differences in the recovery of lactulose (P=0.647) or rhamnose (P=0.889), or in the lactulose/rhamnose ratio, compared with those under conditions of increased permeability alone (P=0.68). However, lactulose recovery was significantly lower (P=0.025) after intravenous administration of a high dose of the sugars. There was no significant difference in urinary rhamnose recovery (P=0.575) between the high and the regular doses. This resulted in a significantly lower lactulose/rhamnose ratio (P=0.021) after intravenous administration of a high dose, compared with a regular dose, of the sugars. In conclusion, the assumption that post-absorptive processes do not influence the outcome of the lactulose/rhamnose permeability test appears not to be valid. PMID- 10677395 TI - Low paraoxonase activity in type II diabetes mellitus complicated by retinopathy. AB - Human serum paraoxonase 1 (PON1) is located on high-density lipoprotein and has been implicated in the detoxification of organophosphates, and possibly in the prevention of lipid peroxidation of low-density lipoprotein. PON1 has two genetic polymorphisms, both due to amino acid substitutions: one involving glutamine (Q genotype) and arginine (R genotype) at position 192, and the other involving leucine (L genotype) and methionine (M genotype) at position 55. We investigated the effects of these polymorphisms, and of a polymorphism of the PON2 gene at position 310 (Cys/Ser; C and S genotypes respectively), on serum PON1 activity and concentration, plasma lipids and lipoproteins and glycaemic control in 93 individuals with type II diabetes with no complications and in 101 individuals with type II diabetes with retinopathy. Serum PON1 activity in the group with no complications [median 164.1 nmol.min(-1).ml(-1) (range 8.0-467.8)] was significantly higher than in the group with retinopathy [113.4 nmol. min(-1).ml( 1) (3.0-414.6)] (P<0.001), but the serum PON1 concentration was not different between the groups. The gene frequencies of the PON1-55 and PON1-192 polymorphisms and of the PON2-310 polymorphism were not different between the study populations. The PON1-55 and PON1-192 polymorphisms affected PON1 activity in the way described in a previous study of a control group and subjects with type II diabetes. The PON2-310 polymorphism also significantly affected serum PON1. PON1 activity was significantly higher in individuals with the PON2-310 CC genotype in both groups with type II diabetes, and the PON1 concentration was significantly higher in PON2-310 CC homozygotes with no complications than in the group with retinopathy. Neither the PON1-55 nor the PON1-192 polymorphism was correlated with the serum lipid or lipoprotein concentration in either group. In the group with retinopathy (but not the group with no complications), all three PON polymorphisms were correlated with glycaemic control, which was worse for the PON1-55 genotypes in the order MM>LM>LL (P=0.0032), for the PON1-192 genotypes in the order RR>QR>QQ (P=0.011) and for the PON2-310 genotypes in the order CC>CS>SS (P=0.010). Low serum PON1 activity in retinopathy may be related to an increased tendency for lipid peroxidation. Our findings thus raise the possibility that, in retinopathy, the PON2 gene may influence PON1, and that an inter-relationship between the PON1 and PON2 genes may influence glycaemic control in subjects with type II diabetes complicated by retinopathy. PMID- 10677396 TI - Percutaneous myocardial laser revascularisation. PMID- 10677397 TI - Five years of percutaneous transluminal septal myocardial ablation. PMID- 10677398 TI - Stamps in cardiology. Acupuncture anaesthesia for open heart surgery. PMID- 10677399 TI - Is there a role for renin profiling in selecting chronic heart failure patients for ACE inhibitor treatment? AB - BACKGROUND: It remains uncertain whether angiotensin converting enzyme (ACE) inhibitors benefit all heart failure patients or just those with renin angiotensin-aldosterone system (RAAS) activation. OBJECTIVE: To determine whether the response to an ACE inhibitor, assessed by urine sodium excretion, was different in patients with low renin versus those with high renin. DESIGN: Plasma renin activity (PRA) was measured in 38 patients with stable chronic heart failure (21 male, 17 female; mean (SD) age 71 (6) years, range 59-82 years) on chronic diuretic treatment alone. They were divided into three groups: low (PRA 5, n = 13). The effect of ACE inhibition was then assessed on diuretic induced natriuresis with respect to renin status. RESULTS: There were no significant differences in age and sex distribution between the groups. Plasma angiotensin II and aldosterone increased serially from low to high renin groups, while 24 h urinary sodium concentrations fell from low to high renin groups (low PRA, 96.7 (39.5); normal PRA, 90.4 (26.7); high PRA, 66. 3 (18.9) mmol/l; p = 0.033), despite a higher diuretic dose in the high renin group. This blunted natriuretic effect of loop diuretics was caused by RAAS activation, which could partly be reversed by ACE inhibition. ACE inhibitors increased natriuresis by 22% in the high renin group (p = 0.029), but had no effect in the normal and low renin groups. Within the low renin group, five of the 11 patients had persistently low renin levels despite ACE inhibition. There was a non-significant reduction in natriuresis (-9.6%, p = 0.335) following ACE inhibition in this subgroup of patients. CONCLUSIONS: About one third of heart failure patients in our study had low renin status and a non-activated RAAS, despite diuretic treatment. ACE inhibitors did not alter natriuresis significantly in this subgroup of patients, and enhanced natriuresis only in patients with high renin. There is thus tentative support for renin profiling in targeting ACE inhibitors to the most deserving, by showing that short term sodium retention does not occur in low renin patients if ACE inhibitors are withdrawn. PMID- 10677400 TI - Images in cardiology. Perivalvar abscess of the mitral valve annulus with perforation owing to infective endocarditis. PMID- 10677401 TI - Effect of verapamil on systolic and diastolic coronary blood flow velocity in asymptomatic and mildly symptomatic patients with hypertrophic cardiomyopathy. AB - OBJECTIVE: To assess non-invasively the effect of verapamil treatment on coronary blood flow velocity in asymptomatic and mildly symptomatic patients with hypertrophic cardiomyopathy. DESIGN: High frequency transthoracic Doppler echocardiography was used to compare resting phasic coronary blood flow velocity before and after a one month period of verapamil treatment in 17 patients (14 men and three women) with non-obstructive hypertrophic cardiomyopathy. Eighteen healthy subjects formed an age and sex matched control group. Systolic and diastolic coronary blood flow velocity was measured in the distal portion of left anterior descending coronary artery using high frequency transthoracic Doppler echocardiography. Blood flow velocity before and after verapamil was compared in the patients with cardiomyopathy and with the results in the control group. RESULTS: Compared with the controls, patients with hypertrophic cardiomyopathy had increased diastolic coronary blood flow velocity (41.8 (8.1) v 59.9 (21.9) cm/s, p < 0.01) and a lower mean systolic coronary blood flow velocity (18.7 (10.8) v -11.2 (27.5) cm/s, p < 0. 01) before verapamil treatment. A backward pattern of systolic flow, manifested by negative values of coronary blood flow velocity, was recorded in eight of the patients, while no negative values were found in the controls. After verapamil treatment the retrograde systolic blood flow was restored to an anterograde pattern in only one patient. The mean value of systolic coronary blood flow velocity did not change significantly and remained lower than the systolic forward flow velocity in the controls (-3.6 (31.8) v 18.7 (10.8) cm/s, p < 0.05). However, diastolic coronary blood flow velocity decreased significantly after verapamil (59.9 (21.9) v 50.7 (19.5) cm/s p < 0.05), reaching a level comparable with that in the controls (50.7 (19.5) v 41.8 (8.1) cm/s, p > 0.05). CONCLUSIONS: In contrast to healthy subjects, in non obstructive hypertrophic cardiomyopathy the systolic pattern of coronary blood flow was heterogeneous (both retrograde and anterograde), and diastolic coronary blood flow velocity was abnormally increased, despite a lack of significant symptoms. Verapamil treatment did not restore the forward pattern of systolic blood flow but decreased diastolic blood flow velocity to a level comparable with that in healthy subjects. PMID- 10677402 TI - Racial variation in cardiovascular morbidity and mortality in essential hypertension. AB - OBJECTIVES: To perform a longitudinal comparison of morbidity and mortality among white, south Asian and Afro-Caribbean hypertensive patients in relation to baseline demographic characteristics and clinic and ambulatory blood pressure variables. DESIGN: Observational follow up study. SETTING: District general hospital and community setting in Harrow, England. PATIENTS: 528 white, 106 south Asian, and 54 Afro-Caribbean subjects with essential hypertension who had undergone 24 hour ambulatory intra-arterial blood pressure monitoring. INTERVENTIONS: Follow up for assessment of all cause morbidity and mortality over a mean (SD) of 9.2 (4.1) years. MAIN OUTCOME MEASURES: Non-cardiovascular death, coronary death, cerebrovascular death, peripheral vascular death, non-fatal myocardial infarction, non-fatal stroke, coronary revascularisation. RESULTS: South Asians had the highest all cause event rate of 3.46, compared with 2.50 (NS) and 0.90 (p = 0.002) events/100 patient-years for whites and Afro Caribbeans, respectively. This was because of an excess of coronary events (2.86 v 1.32 events/100 patient-years in south Asians v whites, respectively; p = 0.002). Age (p < 0.001), sex (p < 0.001), race (south Asians : whites, hazard ratio 1.79; p = 0.008), diabetes (p = 0.05), previous history of cardiovascular disease (p < 0.001), and 24 hour ambulatory systolic blood pressure (p = 0.006) were independent predictors of time to a first event. Clinic blood pressure did not provide additional prognostic information. CONCLUSIONS: South Asian origin was an independent predictor of all cause events, mainly because of an excess of coronary events in this group. Ambulatory but not clinic blood pressure was of additional value in predicting subsequent morbidity and mortality. PMID- 10677403 TI - Blood pressure, arterial compliance, and left ventricular mass: no relation to small size at birth in south Indian adults. AB - OBJECTIVE: To determine whether reduced fetal growth leads to raised blood pressure, reduced arterial compliance, and increased left ventricular mass in an Indian population. DESIGN: A retrospective cohort study of men and women (age range 40-61 years) whose weight, length, and head circumference at birth were recorded. SETTING: The Holdsworth Memorial Hospital, Mysore, South India. SUBJECTS: 435 men and women born in the hospital between 1934 and 1953. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressures; compliance in four arterial segments derived from pulse wave velocity, measured by a non-invasive optical method; and left ventricular mass measured using M mode echocardiography. RESULTS: Small size at birth was not associated with increased adult blood pressure or left ventricular mass, or with reduced arterial compliance. Systolic blood pressure and left ventricular mass were higher in subjects who were greater in length at birth, rising by 1.64 mm Hg (95% confidence interval (CI) -0.08 to +3.37 mm Hg) and 1.63 g/m(2) (95% CI 0.13 to 3.13 g/m(2)), respectively, per one inch (2.5 cm) increase in birth length, independently of adult size. Arterial compliance was reduced in people whose mothers were lighter and had smaller pelvic (external conjugate) diameters. CONCLUSIONS: The higher prevalence of coronary heart disease in Indian men and women of lower birth weight, shown in an earlier study of the same cohort, cannot be explained by changes in blood pressure, arterial compliance, and left ventricular mass. The association of raised blood pressure and left ventricular mass with longer birth length suggests that the way in which the intrauterine environment influences coronary heart disease differs between Indian and Western populations. PMID- 10677404 TI - Images in cardiology. Growth and the implantable cardioverter defibrillator. PMID- 10677405 TI - Influence of hypertension, left ventricular hypertrophy, and left ventricular systolic dysfunction on plasma N terminal proBNP. AB - OBJECTIVES: To examine the relation between plasma concentration of the N terminal of the precursor of brain natriuretic peptide (NT proBNP), left ventricular hypertrophy (LVH), and left ventricular systolic dysfunction (LVSD) in patients with a history of hypertension. DESIGN: Prospective study. SETTING: Teaching hospital based study. PATIENTS: NT proBNP concentrations were determined in five groups of individuals. Group 1: 15 echocardiographic normal controls; group 2: 22 patients with hypertension, normal left ventricular systolic function, and no LVH; group 3: 24 patients with hypertension, normal left ventricular systolic function, and LVH; group 4: 13 patients with history of hypertension, no history of ischaemic heart disease, and left ventricular wall motion index (WMI) between 1.9-1.3; and group 5:17 patients with a history of hypertension, no history of ischaemic heart disease, and WMI < 1.2. RESULTS: Median (range) NT proBNP concentrations (in fmol/ml) for groups 1-5, respectively, were: 129.4 (53.6-159.7), 147.4 (54.3-730. 5), 137.1 (35.8-403.9), 356.7 (124.4-934.4), and 493.5 (248.9-909). Mean log NT proBNP differed among all five groups (p < 0.0001), and between groups 4 and 5 versus groups 1-3 (p < 0.0001), and group 4 versus group 5 (p = 0.02) only. CONCLUSIONS: The results suggest that the presence of hypertension with or without LVH (and normal left ventricular systolic function) does not affect NT proBNP concentrations. Moreover, there is a significant rise in NT proBNP only when LVSD develops in hypertension. Thus, NT proBNP remains a useful diagnostic aid for LVSD, even in hypertensive patients. PMID- 10677406 TI - Prolonged left ventricular dysfunction occurs in patients with coronary artery disease after both dobutamine and exercise induced myocardial ischaemia. AB - OBJECTIVE: To determine whether pharmacological stress leads to prolonged but reversible left ventricular dysfunction in patients with coronary artery disease, similar to that seen after exercise. DESIGN: A randomised crossover study of recovery time of systolic and diastolic left ventricular function after exercise and dobutamine induced ischaemia. SUBJECTS: 10 patients with stable angina, angiographically proven coronary artery disease, and normal left ventricular function. INTERVENTIONS: Treadmill exercise and dobutamine stress were performed on different days. Quantitative assessment of systolic and diastolic left ventricular function was performed using transthoracic echocardiography at baseline and at regular intervals after each test. RESULTS: Both forms of stress led to prolonged but reversible systolic and diastolic dysfunction. There was no difference in the maximum double product (p = 0.53) or ST depression (p = 0.63) with either form of stress. After exercise, ejection fraction was reduced at 15 and 30 minutes compared with baseline (mean (SEM), -5.6 (1.5)%, p < 0.05; and 6.1 (2.2)%, p < 0. 01), and at 30 and 45 minutes after dobutamine (-10.8 (1.8)% and -5. 5 (1.8)%, both p < 0.01). Regional analysis showed a reduction in the worst affected segment 15 and 30 minutes after exercise (-27.9 (7.2)% and -28.6 (5.7)%, both p < 0.01), and at 30 minutes after dobutamine (-32 (5.3)%, p < 0.01). The isovolumic relaxation period was prolonged 45 minutes after each form of stress (p < 0.05). CONCLUSIONS: In patients with coronary artery disease, dobutamine induced ischaemia results in prolonged reversible left ventricular dysfunction, presumed to be myocardial stunning, similar to that seen after exercise. Dobutamine induced ischaemia could therefore be used to study the pathophysiology of this phenomenon further in patients with coronary artery disease. PMID- 10677407 TI - Images in cardiology. Abnormal ventricular conduction following dothiepin overdose simulating acute myocardial infarction. PMID- 10677408 TI - Patients with uncomplicated coronary artery disease have reduced heart rate variability mainly affecting vagal tone. AB - AIM: To investigate whether uncomplicated chronic coronary artery disease causes changes in heart rate variability and if so, whether the heart rate variability pattern is different from that described in patients with acute myocardial infarction. METHODS: Heart rate variability was studied in 65 patients with angina who had no previous myocardial infarcts, no other diseases, and were on no drug that could influence the sinus node. Results were compared with 33 age matched healthy subjects. The diagnosis of coronary artery disease in angina patients was established by coronary angiography in 58, by thallium scintigraphy in six, and by exercise test only in one. Patients and controls were Holter monitored 24 hours outside hospital, and heart rate variability was calculated in the frequency domain as global power (GP: 0.01-1.00 Hz), low frequency peak (LF: 0. 04-0.15 Hz), high frequency peak (HF: 0.15-0.40 Hz), LF/HF in ms(2), and in the time domain as SDNN (SD of normal RR intervals), SDANN (SD of all five minute mean normal RR intervals), SD (mean of all five minute SDs of mean RR intervals), rMSSD (root mean square of differences of successive normal RR intervals) (all in ms), and pNN50 (proportion of adjacent normal RR intervals differing more than 50 ms from the preceding RR interval) as per cent. RESULTS: The mean age in patients and controls was 60.4 (range 32-81) and 59.1 (32-77) years, respectively (NS), the male/female ratio, 57/65 and 24/33 (NS), and the mean time of Holter monitoring, 23.0 (18-24) and 22.8 (18-24) hours (NS). Mortality in angina patients was 0% (0/65) at one year, 0% (0/56) at two years, and 3% (1/33) at three years. Compared with healthy subjects angina patients showed a reduction in GP (p = 0.007), HF (p = 0.02), LF (p = 0.02), SD (p = 0.02), rMSSD (p = 0.01), and pNN50 (p = 0.01). No significant difference was found in RR, LF/HF, SDNN, or SDANN. CONCLUSIONS: Uncomplicated coronary artery disease without previous acute myocardial infarction was associated with reduced high and low frequency heart rate variability, including vagal tone. SDANN and SDNN, expressing ultra low and very low frequencies which are known to reflect prognosis after acute myocardial infarction, were less affected. This is in agreement with the good prognosis in uncomplicated angina in this study. PMID- 10677409 TI - Effects of cardiac sympathetic innervation on regional wall motion abnormality in patients with long QT syndrome. AB - AIM: To assess the spatial relation between regional cardiac sympathetic innervation and regional ventricular repolarisation indicated by ventricular wall motion abnormality in patients with congenital long QT syndrome. DESIGN: Regional percentage uptake and washout rate of (123)I metaiodobenzylguanidine (MIBG) were measured to assess cardiac sympathetic innervation in septum, anterior wall, lateral wall, and posterior wall. Left ventricular short axis images on echocardiography were digitised to reconstruct digitised M mode echocardiograms, from which left ventricular wall thickness curves were obtained. The wall thickening time (ThT) was defined as the period in which the instantaneous wall thickness exceeded 90% of the maximum wall thickness. The ThT was measured from the ventricular wall thickness curve at the same segments where regional percentage uptake and washout rate of (123)I MIBG were measured. PATIENTS: Seven patients with long QT syndrome. RESULTS: The regional washout rate (mean (SD)) of (123)I MIBG in patients with long QT syndrome was greater in the segments with decreased percentage uptake of (123)I MIBG than in those without (17.4 (10.6)% v 9.7 (16.5)%, p < 0. 03). ThT in segments both with and without decreased percentage uptake of (123)I MIBG was longer than in control subjects (p < 0. 0001). ThT was longer in the segments with decreased percentage uptake of (123)I MIBG than in those without (199 (70) ms v 150 (66) ms, p = 0.0018). CONCLUSIONS: Activation of regional cardiac sympathetic terminals is likely to participate in additional regional prolongation of ventricular repolarisation in patients with long QT syndrome. PMID- 10677410 TI - Acute yellow oleander (Thevetia peruviana) poisoning: cardiac arrhythmias, electrolyte disturbances, and serum cardiac glycoside concentrations on presentation to hospital. AB - OBJECTIVE: To describe the cardiac arrhythmias, electrolyte disturbances, and serum cardiac glycoside levels seen in patients presenting to hospital with acute yellow oleander (Thevetia peruviana) poisoning and to compare these with published reports of digitalis poisoning. DESIGN: Case series. SETTING: Medical wards of Anuradhapura District General Hospital, Sri Lanka, and coronary care unit of the Institute of Cardiology, National Hospital of Sri Lanka, Colombo, the national tertiary referral centre for cardiology. PATIENTS: 351 patients with a history of oleander ingestion. MEASUREMENTS: ECG and blood sample analysis on admission. RESULTS: Most symptomatic patients had conduction defects affecting the sinus node, the atrioventricular (AV) node, or both. Patients showing cardiac arrhythmias that required transfer for specialised management had significantly higher mean serum cardiac glycoside and potassium but not magnesium concentrations. Although there was considerable overlap between groups, those with conduction defects affecting both sinus and AV nodes had significantly higher mean serum cardiac glycoside levels. CONCLUSIONS: Most of these young previously healthy patients had conduction defects affecting the sinus or AV nodes. Relatively few had the atrial or ventricular tachyarrhythmias or ventricular ectopic beats that are typical of digoxin poisoning. Serious yellow oleander induced arrhythmias were associated with higher serum cardiac glycoside concentrations and hyperkalaemia but not with disturbances of magnesium. PMID- 10677411 TI - Long term consequences of regressed coronary aneurysms after Kawasaki disease: vascular wall morphology and function. AB - OBJECTIVES: To investigate the long term consequences of regressed aneurysms after Kawasaki disease, using follow up coronary angiography; to assess the vascular wall morphology at the site of the aneurysms by intravascular ultrasound imaging; and to evaluate the function of the affected vessels using intracoronary infusions of acetylcholine and isosorbide dinitrate. DESIGN: 33 patients were studied, 27 with previous Kawasaki disease and six with congenital heart disease. All Kawasaki disease patients were followed for more than 10 years from disease onset. The 33 patients comprised four groups: group 1 included 13 Kawasaki disease patients with a total of 23 sites of regressed large sized (>/= 4 mm) coronary aneurysms; group 2 included 13 Kawasaki disease patients with 22 sites of regressed small sized (< 4 mm) coronary aneurysms (four patients had sites of both large and small sized aneurysms); group 3 included a further five Kawasaki disease patients with 25 normal coronary angiography sites in the acute stage of Kawasaki disease; and group 4 comprised the six patients with congenital heart disease as controls, with a total of 27 normal coronary angiography sites. During coronary angiography, 15 microg of acetylcholine and 0.5 mg isosorbide dinitrate were infused into the coronary artery. The luminal diameter at the sites was measured using a cine-videodensitometric analyser, to assess the distensibility of the coronary artery wall. RESULTS: Coronary angiography in all 22 patients in groups 1 and 2 and in all the patients in group 3 was normal, with no stenoses and no irregularity of the arterial wall. However, the intravascular ultrasound imaging in groups 1 and 2 showed various degrees of the intimal thickening. In groups 1 and 2, there was significantly more vascular constriction with acetylcholine, and poorer dilatation with isosorbide dinitrate than in groups 3 or 4 (each p < 0.05, respectively). There was no difference between group 3 and group 4 in response to either acetylcholine or isosorbide dinitrate, CONCLUSIONS: There is evidence of persisting abnormal vascular wall morphology and vascular dysfunction at the site of regressed coronary aneurysms in patients with previous Kawasaki disease. These patients should be counselled to avoid potential risk factors for atherosclerosis, and long term follow up is needed into adult life. PMID- 10677412 TI - Antiarrhythmic management and implantable defibrillator use in survivors of prehospital cardiac arrest without myocardial infarction in West Yorkshire. AB - OBJECTIVE: To explore the current use of secondary preventive treatment in survivors of out of hospital cardiac arrest without myocardial infarction (primary ventricular tachycardia/ventricular fibrillation (VT/VF)) in West Yorkshire, and assess the implications of recent studies on the benefits of implantable cardioverter-defibrillators (AICD) in this context. DESIGN: Retrospective analysis of an ambulance service based database of outcome after resuscitation of out of hospital cardiac arrest and the Leeds AICD implantation database. MAIN OUTCOME MEASURES: Mortality, rate of referral for specialist investigation, antiarrhythmic treatment. RESULTS: Twelve month mortality following successful discharge after primary VF arrest was 15%. Of 53 patients with primary VF/VT, 29 apparently did not see a cardiologist during the initial admission. Amiodarone was the most widely used antiarrhythmic agent. Six patients (15%) received an AICD. During the same period 22 patients from the same catchment area received an AICD following an in-hospital cardiac arrest. CONCLUSIONS: Mortality among survivors of non-infarct related prehospital cardiac arrest remains significant, with few patients being referred for specialist investigation. The implementation of recent guidelines on AICD use in cardiac arrest survivors would have resulted in an approximate 60% increase in the total numbers of defibrillators implanted in the West Yorkshire area. PMID- 10677413 TI - Partial left ventriculectomy improves left ventricular end systolic elastance in patients with idiopathic dilated cardiomyopathy. AB - OBJECTIVE: To assess the effect of partial left ventriculectomy (PLV) on estimate of left ventricular end systolic elastance (Ees), arterial elastance, and ventriculoarterial coupling. PATIENTS: 11 patients with idiopathic dilated cardiomyopathy before and two weeks after PLV, and 11 controls. INTERVENTIONS: Single plane left ventricular angiography with simultaneous measurements of femoral artery pressure was performed during right heart pacing before and after load reduction. RESULTS: PLV increased mean (SD) Ees from 0.52 (0.27) to 1.47 (0.62) mm Hg/ml (p = 0.0004). The increase in Ees remained significant after correction for the change in left ventricular mass (p = 0.004) and end diastolic volume (p = 0.048). As PLV had no effect on arterial elastance, ventriculoarterial coupling improved from 3.25 (2.17) to 1.01 (0.93) (p = 0.017), thereby maximising left ventricular stroke work. CONCLUSION: It appears that PLV improves both Ees and ventriculoarterial coupling, thus increasing left ventricular work efficiency. PMID- 10677414 TI - Coronary arterial origins in transposition of the great arteries: factors that affect outcome. A morphological and clinical study. AB - OBJECTIVE: Transfer of the coronary arteries is crucial during the arterial switch operation for transposition, but little attention has been paid to the position of their orifices relative to the valvar sinuses. The objective of this study was to determine the factors which are important for effective transfer and to determine potential surgical significance. DESIGN: Morphological and clinical study. SETTING: Two national centres for neonatal cardiac surgery. PATIENTS: 277 patients with transposition of the great arteries. One group comprised 88 necropsy specimens (ages ranging from 17 weeks of fetal life to 17 years old), and the other comprised 189 children undergoing surgery. The coronary artery orifices were inspected relative to the depth of the aortic sinuses (vertical origin), relative to the commissures between the valvar leaflets (radial origin), and their angle of exit from the aortic wall (angle of origin). The data were compared with the surgical results. RESULTS: In the necropsy specimens, the vertical origin of the arteries was at, or above, the sinutubular junction in 20%, the radial origin was paracommissural in 3%, and the angle of origin was not orthogonal in 7%. Those with high take off and paracommissural origin were all intramural. In the clinical cases, those children with high take off, paracommissural origin or tangential origin had an increased risk at surgery. CONCLUSIONS: In 20% of hearts, high take off, paracommissural orifice, or tangential origin of coronary arteries is found. This may be recognised preoperatively by echocardiography and may cause technical difficulty in transfer during the arterial switch procedure. PMID- 10677415 TI - Percutaneous transluminal septal myocardial ablation for hypertrophic obstructive cardiomyopathy: long term follow up of the first series of 25 patients. AB - OBJECTIVE: To determine the long term outcome in patients treated with percutaneous transluminal septal myocardial ablation (PTSMA) for hypertrophic obstructive cardiomyopathy (HOCM). DESIGN AND SETTING: Observational, single centre study. PATIENTS: 25 patients (13 women, 12 men, mean (SD) age 54.7 (15.0) years) with drug treatment resistant New York Heart Association (NYHA) class 2.8 (0. 6) symptoms attributed to a high left ventricular outflow gradient (LVOTG) and a coronary artery anatomy suitable for intervention. INTERVENTION: PTSMA by injection of 4.1 (2.6) ml of alcohol (96%) into 1.4 (0.6) septal perforator arteries to ablate the hypertrophied interventricular septum. OUTCOME MEASURES: During in-hospital follow up, enzyme rise, the frequency of atrioventricular conduction lesions requiring permanent DDD pacing, and in-hospital mortality were assessed. Long term follow up (30 (4) months, range 24-36 months) included symptoms, echocardiographic measurements of left atrial and left ventricular dimensions and function, and LVOTG. RESULTS: Mean postinterventional creatine kinase rise was 780 (436) U/l. During PTSMA 13 patents developed total heart block, permanent pacing being necessary in five of them. One 86 year old patient died from ventricular fibrillation associated with intensive treatment (beta mimetic and theophylline) for coexistent severe obstructive airway disease. After three months, three patients underwent re-PTSMA because of a dissatisfactory primary result, leading to LVOTG elimination in all of them. During long term follow up, LVOTG showed sustained reduction (3 (6) mm Hg at rest and 12 (19) mm Hg with provocation) associated with stable symptomatic improvement (NYHA class 1.2 (1.0)) and without significant global left ventricular dilatation. CONCLUSIONS: PTSMA is an effective non-surgical technique for reduction of symptoms and LVOTG in HOCM. Prospective, long term observations of larger populations are necessary in order to determine the definitive significance of the procedure. PMID- 10677416 TI - Outcome from balloon induced coronary artery dissection after intracoronary beta radiation. AB - OBJECTIVE: To evaluate the healing of balloon induced coronary artery dissection in individuals who have received beta radiation treatment and to propose a new intravascular ultrasound (IVUS) dissection score to facilitate the comparison of dissection through time. DESIGN: Retrospective study. SETTING: Tertiary referral centre. PATIENTS: 31 patients with stable angina pectoris, enrolled in the beta energy restenosis trial (BERT-1.5), were included. After excluding those who underwent stent implantation, the evaluable population was 22 patients. INTERVENTIONS: Balloon angioplasty and intracoronary radiation followed by quantitative coronary angiography (QCA) and IVUS. Repeat QCA and IVUS were performed at six month follow up. MAIN OUTCOME MEASURES: QCA and IVUS evidence of healing of dissection. Dissection classification for angiography was by the National Heart Lung Blood Institute scale. IVUS proven dissection was defined as partial or complete. The following IVUS defined characteristics of dissection were described in the affected coronary segments: length, depth, arc circumference, presence of flap, and dissection score. Dissection was defined as healed when all features of dissection had resolved. The calculated dose of radiation received by the dissected area in those with healed versus non-healed dissection was also compared. RESULTS: Angiography (type A = 5, B = 7, C = 4) and IVUS proven (partial = 12, complete = 4) dissections were seen in 16 patients following intervention. At six month follow up, six and eight unhealed dissections were seen by angiography (A = 2, B = 4) and IVUS (partial = 7, complete = 1), respectively. The mean IVUS dissection score was 5.2 (range 3-8) following the procedure, and 4.6 (range 3-7) at follow up. No correlation was found between the dose prescribed in the treated area and the presence of unhealed dissection. No change in anginal status was seen despite the presence of unhealed dissection. CONCLUSION: beta radiation appears to alter the normal healing process, resulting in unhealed dissection in certain individuals. In view of the delayed and abnormal healing observed, long term follow up is indicated given the possible late adverse effects of radiation. Although in this cohort no increase in cardiac events following coronary dissections was seen, larger populations are needed to confirm this phenomenon. Stenting of all coronary dissections may be warranted in patients scheduled for brachytherapy after balloon angioplasty. PMID- 10677417 TI - Biocompatibility of phosphorylcholine coated stents in normal porcine coronary arteries. AB - OBJECTIVE: To improve the biocompatibility of stents using a phosphorylcholine coated stent as a form of biomimicry. INTERVENTIONS: Implantation of phosphorylcholine coated (n = 20) and non-coated (n = 21) stents was performed in the coronary arteries of 25 pigs. The animals were killed after five days (n = 6), four weeks (n = 7), and 12 weeks (n = 8), and the vessels harvested for histology, scanning electron microscopy, and morphometry. MAIN OUTCOME MEASURES: Stent performance was assessed by studying early endothelialization, neointima formation, and vessel wall reaction to the synthetic coating. RESULTS: Stent thrombosis did not occur in either group. Morphometry showed no significant differences between the two study groups at any time point. At five days both the coated and non-coated stents were equally well endothelialised (91% v 92%, respectively). At four and 12 weeks there was no difference in intimal thickness between the coated and non-coated stents. Up to 12 weeks postimplant the phosphorylcholine coating was still discernible in the stent strut voids, and did not appear to elicit an adverse inflammatory response. CONCLUSION: In this animal model the phosphorylcholine coating showed excellent blood and tissue compatibility, unlike a number of other polymers tested in a similar setting. Given that the coating was present up to 12 weeks postimplant with no adverse tissue reaction, it may be a potential candidate polymer for local drug delivery. PMID- 10677419 TI - Pacemaker lead related tricuspid stenosis: a report of two cases. AB - Only four cases of tricuspid stenosis related to endocardial pacemaker leads have been reported. Two further cases associated with perforation of a tricuspid valve leaflet by a pacemaker lead are presented: a 46 year old woman and a 60 year old man. It is possible that tricuspid valve disease related to endocardial pacemaker and non-thoracotomy defibrillator leads is underrecognized. Diagnosis requires clinical suspicion and the use of Doppler echocardiography. Recent evidence of fibrosis affecting the tricuspid valve in hearts from patients who have had non thoracotomy defibrillator implants suggests that this problem could be more common in the future. PMID- 10677418 TI - Increased serum neopterin: a marker of coronary artery disease activity in women. AB - OBJECTIVE: To assess whether neopterin concentrations in women with unstable angina differ from those in women with chronic stable angina. DESIGN: Prospective cohort study. SETTING: University hospital in south west London. PATIENTS: 114 consecutive women with angina were studied: 82 had chronic stable angina (typical exertional chest pain, positive exercise ECG testing, and/or abnormal myocardial scintigraphy; symptoms stable for at least three months), and 32 had unstable angina (Braunwald class III). All patients with chronic stable angina (100%) and 18 with unstable angina (56.3%) underwent digital coronary angiography; neopterin concentrations were determined using a commercially available immunoassay. MAIN OUTCOME MEASURES: Major clinical events during one year follow up were readmission with Braunwald's class IIIb unstable angina, non-fatal myocardial infarction, and cardiac death. RESULTS: Major events occurred in 12 women with chronic stable angina (14.6%) and nine women with unstable angina (28.1%). Mean (range) neopterin concentrations were significantly higher in women with unstable angina than in those with chronic stable angina (7.6 (5.1-11.5) nmol/l v 5.9 (4.4 7.5) nmol/l; p = 0.003), even after adjustment for variables which were significantly different on univariate analysis. In women with chronic stable angina, baseline neopterin concentrations were higher in those with cardiac events than in those without events (7.1 (5.9-9.1) nmol/l v 5.7 (3.9-7.3 nmol/l); p = 0.010), even after adjustment for variables with significant differences between both groups on univariate analysis. CONCLUSIONS: On average, women with unstable angina had significantly higher neopterin concentrations than women with chronic stable angina. Women with chronic stable angina with events during follow up had higher neopterin concentrations than those without events. Neopterin concentrations were similar in patients with unstable angina and women with chronic stable angina who developed events. Neopterin concentrations may therefore be a marker of risk in women with coronary artery disease. PMID- 10677420 TI - Abnormal atrial and ventricular repolarisation resembling myocardial injury after tricyclic antidepressant drug intoxication. PMID- 10677421 TI - Myocardial perfusion imaging. PMID- 10677422 TI - The pathophysiology of acute coronary syndromes. PMID- 10677423 TI - Antenatal diagnosis of heart disease. PMID- 10677424 TI - Patterns of protein synthesis and tolerance of anoxia in root tips of maize seedlings acclimated to a low-oxygen environment, and identification of proteins by mass spectrometry. AB - Tolerance of anoxia in maize root tips is greatly improved when seedlings are pretreated with 2 to 4 h of hypoxia. We describe the patterns of protein synthesis during hypoxic acclimation and anoxia. We quantified the incorporation of [(35)S]methionine into total protein and 262 individual proteins under different oxygen tensions. Proteins synthesized most rapidly under normoxic conditions continued to account for most of the proteins synthesized during hypoxic acclimation, while the production of a very few proteins was selectively enhanced. When acclimated root tips were placed under anoxia, protein synthesis was depressed and no "new" proteins were detected. We present evidence that protein synthesis during acclimation, but not during subsequent anoxia, is crucial for acclimation. The complex and quantitative changes in protein synthesis during acclimation necessitate identification of large numbers of individual proteins. We show that mass spectrometry can be effectively used to identify plant proteins arrayed by two-dimensional gel electrophoresis. Of the 48 protein spots analyzed, 46 were identified by matching to the protein database. We describe the expression of proteins involved in a wide range of cellular functions, including previously reported anaerobic proteins, and discuss their possible roles in adaptation of plants to low-oxygen stress. PMID- 10677425 TI - Amino acid transporters are localized to transfer cells of developing pea seeds. AB - To determine the nature and cellular localization of amino acid transport in pea seeds, two cDNA clones belonging to the AAP family of H(+)/amino acid co transporters (PsAAP1 and PsAAP2) were isolated from a cotyledon cDNA library of pea (Pisum sativum L.). Functional expression in the yeast amino acid uptake mutants 22Delta6AAL and 22Delta8AA showed that PsAAP1 mediates transport of neutral, acidic, and basic amino acids. RNA-blot analyses showed that PsAAP1 is expressed in seeds and vegetative organs, including amino acid sinks and sources, whereas PsAAP2 could not be detected. For developing seeds, transcripts of PsAAP1 were detected in coats and cotyledons, with seed coats giving a weak signal. In cotyledons, expression was highest in epidermal-transfer-cell-enriched tissue. RNA in situ hybridization analysis showed that PsAAP1 was predominantly present in epidermal transfer cells forming the outer surface of cotyledons, which abuts the seed coats. Overall, our observations suggest that this transporter, which is localized in transfer cells of cotyledons, might play a role in the uptake of the full spectrum of amino acids released from seed coats. PMID- 10677426 TI - Cell division and subsequent radicle protrusion in tomato seeds are inhibited by osmotic stress but DNA synthesis and formation of microtubular cytoskeleton are not. AB - We studied cell cycle events in embryos of tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds during imbibition in water and during osmoconditioning ("priming") using both quantitative and cytological analysis of DNA synthesis and beta-tubulin accumulation. Most embryonic nuclei of dry, untreated control seeds were arrested in the G(1) phase of the cell cycle. This indicated the absence of DNA synthesis (the S-phase), as confirmed by the absence of bromodeoxyuridine incorporation. In addition, beta-tubulin was not detected on western blots and microtubules were not present. During imbibition in water, DNA synthesis was activated in the radicle tip and then spread toward the cotyledons, resulting in an increase in the number of nuclei in G(2). Concomitantly, beta-tubulin accumulated and was assembled into microtubular cytoskeleton networks. Both of these cell cycle events preceded cell expansion and division and subsequent growth of the radicle through the seed coat. The activation of DNA synthesis and the formation of microtubular cytoskeleton networks were also observed throughout the embryo when seeds were osmoconditioned. However, this pre-activation of the cell cycle appeared to become arrested in the G(2) phase since no mitosis was observed. The pre-activation of cell cycle events in osmoconditioned seeds appeared to be correlated with enhanced germination performance during re imbibition in water. PMID- 10677427 TI - Iron deficiency decreases the Fe(III)-chelate reducing activity of leaf protoplasts. AB - The ferric-chelate reductase (FC-R) activity of mesophyll protoplasts isolated from Fe-sufficient (control) and Fe-deficient sugar beet (Beta vulgaris L.) leaves has been characterized. Measurements were made in an ionic environment similar to that in the apoplastic space of the sugar beet mesophyll cells. The FC R activity of Fe-sufficient and Fe-deficient protoplasts was dependent on light. Fe deficiency decreased markedly the FC-R activity per protoplast surface unit. The optimal pH for the activity of the FC-R in mesophyll protoplasts was in the range 5.5 to 6.0, typical of the apoplastic space. Beyond pH 6.0, the activity of the FC-R in mesophyll protoplasts decreased markedly in both Fe-sufficient and Fe deficient protoplasts. These data suggest that both the intrinsic decrease in FC R activity per protoplast surface and a possible shift in the pH of the apoplastic space could lead to the accumulation of physiologically inactive Fe pools in chlorotic leaves. PMID- 10677428 TI - Differentiation of mucilage secretory cells of the Arabidopsis seed coat. AB - In some plant species, including Arabidopsis, fertilization induces the epidermal cells of the outer ovule integument to differentiate into a specialized seed coat cell type with a unique morphology and containing large quantities of polysaccharide mucilage (pectin). Such seed coat mucilage cells are necessary for neither viability nor germination under normal laboratory conditions. Thus, the Arabidopsis seed coat offers a unique system with which to use genetics to identify genes controlling cell morphogenesis and complex polysaccharide biosynthesis and secretion. As a first step in the application of this system, we have used microscopy to investigate the structure and differentiation of Arabidopsis seed coat mucilage cells, including cell morphogenesis and the synthesis, secretion, and extrusion of mucilage. During seed coat development in Arabidopsis, the epidermal cells of the outer ovule integument grow and differentiate into cells that produce large quantities of mucilage between the primary cell wall and plasma membrane. Concurrent with mucilage production, the cytoplasm is shaped into a column in the center of the cell. Following mucilage secretion the cytoplasmic column is surrounded by a secondary cell wall to form a structure known as the columella. Thus, differentiation of the seed coat mucilage cells involves a highly regulated series of events including growth, morphogenesis, mucilage biosynthesis and secretion, and secondary cell wall synthesis. PMID- 10677429 TI - Hypersensitivity of an Arabidopsis sugar signaling mutant toward exogenous proline application. AB - In transgenic Arabidopsis a patatin class I promoter from potato is regulated by sugars and proline (Pro), thus integrating signals derived from carbon and nitrogen metabolism. In both cases a signaling cascade involving protein phosphatases is involved in induction. Other endogenous genes are also regulated by both Pro and carbohydrates. Chalcone synthase (CHS) gene expression is induced by both, whereas the Pro biosynthetic Delta(1)-pyrroline-5-carboxylate synthetase (P5CS) is induced by high Suc concentrations but repressed by Pro, and Pro dehydrogenase (ProDH) is inversely regulated. The mutant rsr1-1, impaired in sugar dependent induction of the patatin promoter, is hypersensitive to low levels of external Pro and develops autofluorescence and necroses. Toxicity of Pro can be ameliorated by salt stress and exogenously supplied metabolizable carbohydrates. The rsr1-1 mutant shows a reduced response regarding sugar induction of CHS and P5CS expression. ProDH expression is de-repressed in the mutant but still down-regulated by sugar. Pro toxicity seems to be mediated by the degradation intermediate Delta(1)-pyrroline-5-carboxylate. Induction of the patatin promoter by carbohydrates and Pro, together with the Pro hypersensitivity of the mutant rsr1-1, demonstrate a new link between carbon/nitrogen and stress responses. PMID- 10677430 TI - Expression and localization of nitrilase during symptom development of the clubroot disease in Arabidopsis. AB - The expression of nitrilase in Arabidopsis during the development of the clubroot disease caused by the obligate biotroph Plasmodiophora brassicae was investigated. A time course study showed that only during the exponential growth phase of the clubs was nitrilase prominently enhanced in infected roots compared with controls. NIT1 and NIT2 are the nitrilase isoforms predominantly expressed in clubroot tissue, as shown by investigating promoter-beta-glucuronidase fusions of each. Two peaks of beta-glucuronidase activity were visible: an earlier peak (21 d post inoculation) consisting only of the expression of NIT1, and a second peak at about 32 d post inoculation, which predominantly consisted of NIT2 expression. Using a polyclonal antibody against nitrilase, it was shown that the protein was mainly found in infected cells containing sporulating plasmodia, whereas in cells of healthy roots and in uninfected cells of inoculated roots only a few immunosignals were detected. To determine which effect a missing nitrilase isoform might have on symptom development, the P. brassicae infection in a nitrilase mutant (nit1-3) of Arabidopsis was investigated. As a comparison, transgenic plants overexpressing NIT2 under the control of the cauliflower mosaic virus 35S promoter were studied. Root galls were smaller in nit1-3 plants compared with the wild type. The phenotype of smaller clubs in the mutant was correlated with a lower free indole-3-acetic acid content in the clubs compared with the wild type. Overexpression of nitrilase did not result in larger clubs compared with the wild type. The putative role of nitrilase and auxins during symptom development is discussed. PMID- 10677431 TI - Cloning and functional characterization of a constitutively expressed nitrate transporter gene, OsNRT1, from rice. AB - Elucidating how rice (Oryza sativa) takes up nitrate at the molecular level could help improve the low recovery rate (<50%) of nitrogen fertilizer in rice paddies. As a first step toward that goal, we have cloned a nitrate transporter gene from rice called OsNRT1. OsNRT1 is a new member of a growing transporter family called PTR, which consists not only of nitrate transporters from higher plants that are homologs of the Arabidopsis CHL1 (AtNRT1) protein, but also peptide transporters from a wide variety of genera including animals, plants, fungi, and bacteria. However, despite the fact that OsNRT1 shares a higher degree of sequence identity with the two peptide transporters from plants (approximately 50%) than with the nitrate transporters (approximately 40%) of the PTR family, no peptide transport activity was observed when OsNRT1 was expressed in either Xenopus oocytes or yeast. Furthermore, contrasting the dual-affinity nitrate transport activity of CHL1, OsNRT1 displayed only low-affinity nitrate transport activity in Xenopus oocytes, with a K(m) value of approximately 9 mM. Northern-blot and in situ hybridization analysis indicated that OsNRT1 is constitutively expressed in the most external layer of the root, epidermis and root hair. These data strongly indicate that OsNRT1 encodes a constitutive component of a low-affinity nitrate uptake system for rice. PMID- 10677432 TI - Toward a functional catalog of the plant genome. A survey of genes for lipid biosynthesis. AB - Public databases now include vast amounts of recently acquired DNA sequences that are only partially annotated and, furthermore, are often annotated by automated methods that are subject to errors. Maximum information value of these databases can be derived only by further detailed analyses that frequently require careful examination of records in the context of biological functions. In this study we present an example of such an analysis focused on plant glycerolipid synthesis. Public databases were searched for sequences corresponding to 65 plant polypeptides involved in lipid metabolism. Comprehensive search results and analysis of genes, cDNAs and expressed sequence tags (ESTs) are available online (http://www.canr.msu.edu/lgc). Multiple alignments provided a method to estimate the number of genes in gene families. Further analysis of sequences allowed us to tentatively identify several previously undescribed genes in Arabidopsis. For example, two genomic sequences were identified as candidates for the palmitate specific monogalactosyldiacylglycerol desaturase (FAD5). A candidate genomic sequence for 3-ketoacyl-acyl-carrier protein (ACP) synthase involved in mitochondrial fatty acid biosynthesis was also identified. Biotin carboxyl carrier protein (BCCP) in Arabidopsis is encoded by at least two genes, but the most abundant BCCP transcript so far has not been characterized. The large number (>165,000) of plant ESTs also provides an opportunity to perform "digital northern" comparisons of gene expression levels across many genes. EST abundance in general correlated with biochemical and flux characteristics of the enzymes in Arabidopsis leaf tissue. In a few cases, statistically significant differences in EST abundance levels were observed for enzymes that catalyze similar reactions in fatty acid metabolism. For example, ESTs for the FatB acyl-ACP thioesterase occur 21 times compared with 7 times for FatA acyl-ACP thioesterase, although flux through the FatA reaction is several times higher than through FatB. Such comparisons may provide initial clues toward previously undescribed regulatory phenomena. The abundance of ESTs for ACP compared with that of stearoyl-ACP desaturase and FatB acyl-ACP thioesterase suggests that concentrations of some enzymes of fatty acid synthesis may be higher than their acyl-ACP substrates. PMID- 10677433 TI - Influence of the testa on seed dormancy, germination, and longevity in Arabidopsis. AB - The testa of higher plant seeds protects the embryo against adverse environmental conditions. Its role is assumed mainly by controlling germination through dormancy imposition and by limiting the detrimental activity of physical and biological agents during seed storage. To analyze the function of the testa in the model plant Arabidopsis, we compared mutants affected in testa pigmentation and/or structure for dormancy, germination, and storability. The seeds of most mutants exhibited reduced dormancy. Moreover, unlike wild-type testas, mutant testas were permeable to tetrazolium salts. These altered dormancy and tetrazolium uptake properties were related to defects in the pigmentation of the endothelium and its neighboring crushed parenchymatic layers, as determined by vanillin staining and microscopic observations. Structural aberrations such as missing layers or a modified epidermal layer in specific mutants also affected dormancy levels and permeability to tetrazolium. Both structural and pigmentation mutants deteriorated faster than the wild types during natural aging at room temperature, with structural mutants being the most strongly affected. PMID- 10677434 TI - Gibberellin requirement for Arabidopsis seed germination is determined both by testa characteristics and embryonic abscisic acid. AB - The mechanisms imposing a gibberellin (GA) requirement to promote the germination of dormant and non-dormant Arabidopsis seeds were analyzed using the GA-deficient mutant ga1, several seed coat pigmentation and structure mutants, and the abscisic acid (ABA)-deficient mutant aba1. Testa mutants, which exhibit reduced seed dormancy, were not resistant to GA biosynthesis inhibitors such as tetcyclacis and paclobutrazol, contrarily to what was found before for other non dormant mutants in Arabidopsis. However, testa mutants were more sensitive to exogenous GAs than the wild-types in the presence of the inhibitors or when transferred to a GA-deficient background. The germination capacity of the ga1-1 mutant could be integrally restored, without the help of exogenous GAs, by removing the envelopes or by transferring the mutation to a tt background (tt4 and ttg1). The double mutants still required light and chilling for dormancy breaking, which may indicate that both agents can have an effect independently of GA biosynthesis. The ABA biosynthesis inhibitor norflurazon was partially efficient in releasing the dormancy of wild-type and mutant seeds. These results suggest that GAs are required to overcome the germination constraints imposed both by the seed coat and ABA-related embryo dormancy. PMID- 10677435 TI - Purification and characterization of barley dipeptidyl peptidase IV. AB - Barley (Hordeum vulgare L.) storage proteins, which have a high content of proline (Pro) and glutamine, are cleaved by cysteine endoproteases to yield peptides with a Pro next to the N-terminal and/or C-terminal amino acid residues. A peptidase cleaving after Xaa-Pro- at the N terminus of peptides was purified from green barley malt. It was identified as a serine-type dipeptidyl peptidase (DPP), based on inhibitor studies, and the nature of the cleavage product. It is a monomeric glycoprotein with an apparent molecular mass of 105 kD (85 kD after deglycosylation), with a pI of 3.55 and a pH optimum at 7.2. Substrate specificity was determined with a series of fluorogenic peptide substrates with the general formula Xaa-Pro-AMC, where Xaa is an unspecified amino acid and AMC is 7-amino-4-methylcoumarin. The best substrates were Xaa = lysine and arginine, while the poorest were Xaa = aspartic acid, phenylalanine, and glutamic acid. The K(m) values ranged from 0.071 to 8.9 microM, compared with values of 9 to 130 microM reported for mammalian DPP IVs. We discuss the possible role of DPP IV in the degradation of small Pro-containing peptides transported from the endosperm to the embryo of the germinating barley grain. PMID- 10677436 TI - Major protein of resting rhizomes of Calystegia sepium (hedge bindweed) closely resembles plant RNases but has no enzymatic activity. AB - The most abundant protein of resting rhizomes of Calystegia sepium (L.) R.Br. (hedge bindweed) has been isolated and its corresponding cDNA cloned. The native protein consists of a single polypeptide of 212 amino acid residues and occurs as a mixture of glycosylated and unglycosylated isoforms. Both forms are derived from the same preproprotein containing a signal peptide and a C-terminal propeptide. Analysis of the deduced amino acid sequence indicated that the C. sepium protein shows high sequence identity and structural similarity with plant RNases. However, no RNase activity could be detected in highly purified preparations of the protein. This apparent lack of activity results most probably from the replacement of a conserved His residue, which is essential for the catalytic activity of plant RNases. Our findings not only demonstrate the occurrence of a catalytically inactive variant of an S-like RNase, but also provide further evidence that genes encoding storage proteins may have evolved from genes encoding enzymes or other biologically active proteins. PMID- 10677437 TI - Nucleoside diphosphate kinase required for coleoptile elongation in rice. AB - Although several nucleoside diphosphate (NDP) kinase genes have been cloned in plants, little is known about the functional significance of this enzyme during plant growth and development. We introduced a chimeric gene encoding an antisense RNA of NDP kinase under the control of the Arabidopsis heat shock protein HSP81-1 promoter into rice (Oryza sativa L.) plants using the Agrobacterium tumefaciens transformation system. The expression of antisense RNA down-regulated the accumulation of mRNA, resulting in reduced enzyme activity even under the standard growth temperature (25 degrees C) in transgenic plants. Following heat shock treatment (37 degrees C), NDP kinase activities in some transgenic rice plants were more reduced than those grown under 25 degrees C. The comparison of the coleoptile growth under submersion showed that cell elongation process was inhibited in antisense NDP kinase transgenic plants, suggesting that an altered guanine nucleotide level may be responsible for the processes. PMID- 10677438 TI - Interaction of root gravitropism and phototropism in Arabidopsis wild-type and starchless mutants. AB - Root gravitropism in wild-type Arabidopsis and in two starchless mutants, pgm1-1 and adg1-1, was evaluated as a function of light position to determine the relative strengths of negative phototropism and of gravitropism and how much phototropism affects gravitropic measurements. Gravitropism was stronger than phototropism in some but not all light positions in wild-type roots grown for an extended period, indicating that the relationship between the two tropisms is more complex than previously reported. Root phototropism significantly influenced the time course of gravitropic curvature and the two measures of sensitivity. Light from above during horizontal exposure overestimated all three parameters for all three genotypes except the wild-type perception time. At the irradiance used (80 micromol m(-2) s(-1)), the shortest periods of illumination found to exaggerate gravitropism were 45 min of continuous illumination and 2-min doses of intermittent illumination. By growing roots in circumlateral light or by gravistimulating in the dark, corrected values were obtained for each gravitropic parameter. Roots of both starchless mutants were determined to be about three times less sensitive than prior estimates. This study demonstrates the importance of accounting for phototropism in the design of root gravitropism experiments in Arabidopsis. PMID- 10677439 TI - Phenylarsine oxide inhibits the fusicoccin-induced activation of plasma membrane H(+)-ATPase. AB - To investigate the mechanism by which fusicoccin (FC) induces the activation of the plasma membrane (PM) H(+)-ATPase, we used phenylarsine oxide (PAO), a known inhibitor of protein tyrosine-phosphatases. PAO was supplied in vivo in the absence or presence of FC to radish (Raphanus sativus L.) seedlings and cultured Arabidopsis cells prior to PM extraction. Treatment with PAO alone caused a slight decrease of PM H(+)-ATPase activity and, in radish, a decrease of PM associated 14-3-3 proteins. When supplied prior to FC, PAO drastically inhibited FC-induced activation of PM H(+)-ATPase, FC binding to the PM, and the FC-induced increase of the amount of 14-3-3 associated with the PM. On the contrary, PAO was completely ineffective on all of the above-mentioned parameters when supplied after FC. The H(+)-ATPase isolated from PAO-treated Arabidopsis cells maintained the ability to respond to FC if supplied with exogenous, nonphosphorylated 14-3-3 proteins. Altogether, these results are consistent with a model in which the dephosphorylated state of tyrosine residues of a protein(s), such as 14-3-3 protein, is required to permit FC-induced association between the 14-3-3 protein and the PM H(+)-ATPase. PMID- 10677440 TI - The gene pat-2, which induces natural parthenocarpy, alters the gibberellin content in unpollinated tomato ovaries. AB - We investigated the role of gibberellins (GAs) in the effect of pat-2, a recessive mutation that induces facultative parthenocarpic fruit development in tomato (Lycopersicon esculentum Mill.) using near-isogenic lines with two different genetic backgrounds. Unpollinated wild-type Madrigal (MA/wt) and Cuarenteno (CU/wt) ovaries degenerated, but GA(3) application induced parthenocarpic fruit growth. On the contrary, parthenocarpic growth of MA/pat-2 and CU/pat-2 fruits, which occurs in the absence of pollination and hormone application, was not affected by GA(3). Pollinated MA/wt and parthenocarpic MA/pat-2 ovary development was negated by paclobutrazol, and this inhibitory effect was counteracted by GA(3). The main GAs of the early-13-hydroxylation pathway (GA(1), GA(3), GA(8), GA(19), GA(20), GA(29), GA(44), GA(53), and, tentatively, GA(81)) and two GAs of the non-13-hydroxylation pathway (GA(9) and GA(34)) were identified in MA/wt ovaries by gas chromatography-selected ion monitoring. GAs were quantified in unpollinated ovaries at flower bud, pre anthesis, and anthesis. In unpollinated MA/pat-2 and CU/pat-2 ovaries, the GA(20) content was much higher (up to 160 times higher) and the GA(19) content was lower than in the corresponding non-parthenocarpic ovaries. The application of an inhibitor of 2-oxoglutarate-dependent dioxygenases suggested that GA(20) is not active per se. The pat-2 mutation may increase GA 20-oxidase activity in unpollinated ovaries, leading to a higher synthesis of GA(20), the precursor of an active GA. PMID- 10677441 TI - Basipetal auxin transport is required for gravitropism in roots of Arabidopsis. AB - Auxin transport has been reported to occur in two distinct polarities, acropetally and basipetally, in two different root tissues. The goals of this study were to determine whether both polarities of indole-3-acetic acid (IAA) transport occur in roots of Arabidopsis and to determine which polarity controls the gravity response. Global application of the auxin transport inhibitor naphthylphthalamic acid (NPA) to roots blocked the gravity response, root waving, and root elongation. Immediately after the application of NPA, the root gravity response was completely blocked, as measured by an automated video digitizer. Basipetal [(3)H]IAA transport in Arabidopsis roots was inhibited by NPA, whereas the movement of [(14)C]benzoic acid was not affected. Inhibition of basipetal IAA transport by local application of NPA blocked the gravity response. Inhibition of acropetal IAA transport by application of NPA at the root-shoot junction only partially reduced the gravity response at high NPA concentrations. Excised root tips, which do not receive auxin from the shoot, exhibited a normal response to gravity. The Arabidopsis mutant eir1, which has agravitropic roots, exhibited reduced basipetal IAA transport but wild-type levels of acropetal IAA transport. These results support the hypothesis that basipetally transported IAA controls root gravitropism in Arabidopsis. PMID- 10677442 TI - The role of chloroplast electron transport and metabolites in modulating Rubisco activity in tobacco. Insights from transgenic plants with reduced amounts of cytochrome b/f complex or glyceraldehyde 3-phosphate dehydrogenase. AB - Leaf metabolites, adenylates, and Rubisco activation were studied in two transgenic tobacco (Nicotiana tabacum L. cv W38) types. Plants with reduced amounts of cytochrome b/f complex (anti-b/f) have impaired electron transport and a low transthylakoid pH gradient that restrict ATP and NADPH synthesis. Plants with reduced glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH) have a decreased capacity to use ATP and NADPH in carbon assimilation. The activation of the chloroplast NADP-malate dehydrogenase decreased in anti-b/f plants, indicating a low NADPH/NADP(+) ratio. The whole-leaf ATP/ADP in anti-b/f plants was similar to wild type, while it increased in anti-GAPDH plants. In both plant types, the CO(2) assimilation rates decreased with decreasing ribulose 1, 5 bisphosphate concentrations. In anti-b/f plants, CO(2) assimilation was further compromised by reduced carbamylation of Rubisco, whereas in anti-GAPDH plants the carbamylation remained high even at subsaturating ribulose 1,5-bisphosphate concentrations. We propose that the low carbamylation in anti-b/f plants is due to reduced activity of Rubisco activase. The results suggest that light modulation of activase is not directly mediated via the electron transport rate or stromal ATP/ADP, but some other manifestation of the balance between electron transport and the consumption of its products. Possibilities include the transthylakoid pH gradient and the reduction state of the acceptor side of photosystem I and/or the degree of reduction of the thioredoxin pathway. PMID- 10677443 TI - Redox control of psbA gene expression in the cyanobacterium Synechocystis PCC 6803. Involvement of the cytochrome b(6)/f complex. AB - We investigated the role of the redox state of the photosynthetic and respiratory electron transport chains on the regulation of psbA expression in Synechocystis PCC 6803. Different means to modify the redox state of the electron carriers were used: (a) dark to oxidize the whole electron transport chain; (b) a shift from dark to light to induce its reduction; (c) the chemical interruption of the electron flow at different points to change the redox state of specific electron carriers; and (d) the presence of glucose to maintain a high reducing power in darkness. We show that changes in the redox state of the intersystem electron transport chain induce modifications of psbA transcript production and psbA mRNA stability. Reduction of the intersystem electron carriers activates psbA transcription and destabilizes the mRNA, while their oxidation induces a decrease in transcription and a stabilization of the transcript. Furthermore, our data suggest that the redox state of one of the electron carriers between the plastoquinone pool and photosystem I influences not only the expression of the psbA gene, but also that of other two photosynthetic genes, psaE and cpcBA. As a working hypothesis, we propose that the occupancy of the Q(0) site in the cytochrome b(6)/f complex may be involved in this regulation. PMID- 10677444 TI - Regulatory role of the N terminus of the vacuolar calcium-ATPase in cauliflower. AB - The vacuolar calmodulin (CaM)-stimulated Ca(2+)-ATPase, BCA1p, in cauliflower (Brassica oleracea) has an extended N terminus, which was suggested to contain a CaM-binding domain (S. Malmstrom, P. Askerlund, M.G. Palmgren [1997] FEBS Lett 400: 324-328). The goal of the present study was to determine the role of the N terminus in regulating BCA1p. Western analysis using three different antisera showed that the N terminus of BCA1p is cleaved off by trypsin and that the N terminus contains the CaM-binding domain. Furthermore, the expressed N terminus binds CaM in a Ca(2+)-dependent manner. A synthetic peptide corresponding to the CaM-binding domain of BCA1p (Ala-19 to Leu-43) strongly inhibited ATP-dependent Ca(2+) pumping by BCA1p in cauliflower low-density membranes, indicating that the CaM-binding region of BCA1p also has an autoinhibitory function. The expressed N terminus of BCA1p and a synthetic peptide (Ala-19 to Met-39) were good substrates for phosphorylation by protein kinase C. Sequencing of the phosphorylated fusion protein and peptide suggested serine-16 and/or serine-28 as likely targets for phosphorylation. Phosphorylation of serine-28 had no effect on CaM binding to the alanine-19 to methionine-39 peptide. Our results demonstrate the regulatory importance of the N terminus of BCA1p as a target for CaM binding, trypsin cleavage, and phosphorylation, as well as its importance as an autoinhibitory domain. PMID- 10677445 TI - Auxin-regulated genes encoding cell wall-modifying proteins are expressed during early tomato fruit growth. AB - An expansin gene, LeExp2, was isolated from auxin-treated, etiolated tomato (Lycopersicon esculentum cv T5) hypocotyls. LeExp2 mRNA expression was restricted to the growing regions of the tomato hypocotyl and was up-regulated during incubation of hypocotyl segments with auxin. The pattern of expression of LeExp2 was also studied during tomato fruit growth, a developmental process involving rapid cell enlargement. The expression of genes encoding a xyloglucan endotransglycosylase (LeEXT1) and an endo-1, 4-beta-glucanase (Cel7), which, like LeExp2, are auxin-regulated in etiolated hypocotyls (C. Catala, J.K.C. Rose, A.B. Bennett [1997] Plant J 12: 417-426), was also studied to examine the potential for synergistic action with expansins. LeExp2 and LeEXT1 genes were coordinately regulated, with their mRNA accumulation peaking during the stages of highest growth, while Cel7 mRNA abundance increased and remained constant during later stages of fruit growth. The expression of LeExp2, LeEXT1, and Cel7 was undetectable or negligible at the onset of and during fruit ripening, which is consistent with a specific role of these genes in regulating cell wall loosening during fruit growth, not in ripening-associated cell wall disassembly. PMID- 10677446 TI - Intron-mediated enhancement of gene expression independent of unique intron sequences and splicing. AB - Either of the first two introns of the Arabidopsis tryptophan pathway gene PAT1 elevates mRNA accumulation from a PAT1:beta-glucuronidase (GUS) fusion roughly 5 fold without affecting the rate of PAT1:GUS transcription. To further explore the mechanism of this intron-mediated enhancement of gene expression, we wanted to determine whether splicing or specific intron sequences were necessary. In-frame derivatives of PAT1 intron 1, whose splicing was prevented by a point mutation or large deletions, were able to increase mRNA accumulation from a PAT1:GUS fusion, demonstrating that splicing per se is not required. Furthermore, each of a series of introns containing overlapping deletions that together span PAT1 intron 1 increased PAT1:GUS mRNA accumulation as much as the full-length intron did, indicating that all intron sequences are individually dispensable for this phenomenon. These results eliminate the simple idea that this intron stimulates mRNA accumulation via a unique RNA-stabilizing sequence or through the completed act of splicing. However, they are consistent with a possible role for redundant intron sequence elements or an association of the pre-mRNA with the spliceosome. PMID- 10677447 TI - Induction of an extracellular cyclic nucleotide phosphodiesterase as an accessory ribonucleolytic activity during phosphate starvation of cultured tomato cells. AB - During growth under conditions of phosphate limitation, suspension-cultured cells of tomato (Lycopersicon esculentum Mill.) secrete phosphodiesterase activity in a similar fashion to phosphate starvation-inducible ribonuclease (RNase LE), a cyclizing endoribonuclease that generates 2':3'-cyclic nucleoside monophosphates (NMP) as its major monomeric products (T. Nurnberger, S. Abel, W. Jost, K. Glund [1990] Plant Physiol 92: 970-976). Tomato extracellular phosphodiesterase was purified to homogeneity from the spent culture medium of phosphate-starved cells and was characterized as a cyclic nucleotide phosphodiesterase. The purified enzyme has a molecular mass of 70 kD, a pH optimum of 6.2, and an isoelectric point of 8.1. The phosphodiesterase preparation is free of any detectable deoxyribonuclease, ribonuclease, and nucleotidase activity. Tomato extracellular phosphodiesterase is insensitive to EDTA and hydrolyzes with no apparent base specificity 2':3'-cyclic NMP to 3'-NMP and the 3':5'-cyclic isomers to a mixture of 3'-NMP and 5'-NMP. Specific activities of the enzyme are 2-fold higher for 2':3'-cyclic NMP than for 3':5'-cyclic isomers. Analysis of monomeric products of sequential RNA hydrolysis with purified RNase LE, purified extracellular phosphodiesterase, and cleared -Pi culture medium as a source of 3'-nucleotidase activity indicates that cyclic nucleotide phosphodiesterase functions as an accessory ribonucleolytic activity that effectively hydrolyzes primary products of RNase LE to substrates for phosphate-starvation-inducible phosphomonoesterases. Biosynthetical labeling of cyclic nucleotide phopshodiesterase upon phosphate starvation suggests de novo synthesis and secretion of a set of nucleolytic enzymes for scavenging phosphate from extracellular RNA substrates. PMID- 10677448 TI - Nitrogenase activity in Alnus incana root nodules. Responses to O(2) and short term N(2) deprivation. AB - O(2) and host-microsymbiont interactions are key factors affecting the physiology of N(2)-fixing symbioses. To determine the relationship among nitrogenase activity of Frankia-Alnus incana root nodules, O(2) concentration, and short-term N(2) deprivation, intact nodulated roots were exposed to various O(2) pressures (pO(2)) and Ar:O(2) in a continuous flow-through system. Nitrogenase activity (H(2) production) occurred at a maximal rate at 20% O(2). Exposure to short-term N(2) deprivation in Ar:O(2) carried out at either 17%, 21%, or 25% O(2) caused a decline in the nitrogenase activity at 21% and 25% O(2) by 12% and 25%, respectively. At 21% O(2), nitrogenase activity recovered to initial activity within 60 min. The decline rate was correlated with the degree of inhibition of N(2) fixation. Respiration (net CO(2) evolution) decreased in response to the N(2) deprivation at all pO(2) values and did not recover during the time in Ar:O(2). Increasing the pO(2) from 21% to 25% and decreasing the pO(2) from 21% to 17% during the decline further decreased rather than stimulated nitrogenase activity, showing that the decline was not due to O(2) limitation. The decline was possibly due to a temporary disturbance in the supply of reductant to nitrogenase with a partial O(2) inhibition of nitrogenase at 25% O(2). These results are consistent with a fixed O(2) diffusion barrier in A. incana root nodules, and show that A. incana nodules differ from legume nodules in the response of the nitrogenase activity to O(2) and N(2) deprivation. PMID- 10677449 TI - A specific beta-glucosidase-aggregating factor is responsible for the beta glucosidase null phenotype in maize. AB - Maize (Zea mays L.) beta-glucosidase was extracted from shoots of a wild-type (K55) and a "null" (H95) maize genotype. Enzyme activity assays and electrophoretic data showed that extracts from the null genotype had about 10% of the activity present in the normal genotype. Zymograms of the null genotype were devoid of any activity bands in the resolving gel, but had a smeared zone of activity in the stacking gel after native polyacrylamide gel electrophoresis. When extracts were made with buffers containing 0.5% to 2% sodium dodecyl sulfate, the smeared activity zone entered the resolving gel as a distinct band. These data indicated that the null genotypes have beta-glucosidase activity, but the enzyme occurs as insoluble or poorly soluble large quaternary complexes mediated by a beta-glucosidase-aggregating factor (BGAF). BGAF is a 35-kD protein and binds specifically to beta-glucosidase and renders it insoluble during extraction. BGAF also precipitates beta-glucosidase that is added exogenously to supernatant fluids of the null tissue extracts. The specific beta-glucosidase aggregating activity of BGAF is unequivocally demonstrated. These data clearly show that the monogenic inheritance reported for the null alleles at the beta glucosidase gene is actually for the BGAF protein, and BGAF is solely responsible for beta-glucosidase aggregation and insolubility and, thus, the apparent null phenotype. PMID- 10677451 TI - Distinguishing between luminal and localized proton buffering pools in thylakoid membranes. AB - The dual gradient energy coupling hypothesis posits that chloroplast thylakoid membranes are energized for ATP formation by either a delocalized or a localized proton gradient geometry. Localized energy coupling is characterized by sequestered domains with a buffering capacity of approximately 150 nmol H(+) mg( 1) chlorophyll (Chl). A total of 30 to 40 nmol mg(-1) Chl of the total sequestered domain buffering capacity is contributed by lysines with anomolously low pK(a)s, which can be covalently derivatized with acetic anhydride. We report that in thylakoid membranes treated with acetic anhydride, luminal acidification by a photosystem I (duraquinol [DQH(2)] to methyl viologen [MV]) proton pumping partial reaction was nearly completely inhibited, as measured by three separate assays, yet surprisingly, H(+) accumulation still occurred to the significant level of more than 100 nmol H(+) mg Chl(-1), presumably into the sequestered domains. The treatment did not increase the observed rate constant of dark H(+) efflux, nor was electron transport significantly inhibited. These data provide support for the existence of a sequestered proton translocating pathway linking the redox reaction H(+) ion sources with the CF(0) H(+) channel. The sequestered, low-pK(a) Lys groups appear to have a role in the H(+) diffusion process and chemically modifying them blocks the putative H(+) relay system. PMID- 10677450 TI - Nitric oxide modulates the activity of tobacco aconitase. AB - Recent evidence suggests an important role for nitric oxide (NO) signaling in plant-pathogen interactions. Additional elucidation of the role of NO in plants will require identification of NO targets. Since aconitases are major NO targets in animals, we examined the effect of NO on tobacco (Nicotiana tabacum) aconitase. The tobacco aconitases, like their animal counterparts, were inhibited by NO donors. The cytosolic aconitase in animals, in addition to being a key redox and NO sensor, is converted by NO into an mRNA binding protein (IRP, or iron-regulatory protein) that regulates iron homeostasis. A tobacco cytosolic aconitase gene (NtACO1) whose deduced amino acid sequence shared 61% identity and 76% similarity with the human IRP-1 was cloned. Furthermore, residues involved in mRNA binding by IRP-1 were conserved in NtACO1. These results reveal additional similarities between the NO signaling mechanisms used by plants and animals. PMID- 10677453 TI - Exposure to low irradiances favors the synthesis of 9-cis beta, beta-carotene in Dunaliella salina (Teod.). AB - We examined the effect of irradiance on the synthesis of beta-carotene and its isomers by Dunaliella salina. Growth irradiance had a marked effect both on growth of the alga (which was suppressed at both low and high irradiances) and on the accumulation of beta-carotene. The accumulation of beta-carotene but not alpha-carotene was closely linked to an increase in irradiance. Growth at low irradiances (20-50 micromol m(-2) s(-1)) promoted a high ratio of 9-cis to all trans beta-carotene (>2:1), while exposure to high irradiances (200-1,250 micromol m(-2) s(-1)) resulted in a large reduction in this ratio (to <0.45:1). A similar pattern was seen for the geometric isomers of alpha-carotene, with exposure to low irradiance favoring the accumulation of the 9-cis form. The carotenoid biosynthesis inhibitors 4-chloro-5(methylamino)-2-(alpha-alpha-alpha trifluoro-m-tolyl)-3-(sH )-pyridazinone and 2-(4-chlorophenylthio)triethylamine caused the accumulation of the precursors phytoene and lycopene, respectively, in D. salina. High-performance liquid chromatography and infrared analysis showed that phytoene adopted the 15-cis and all-trans forms (as in higher plants), and that lycopene primarily adopted the all-trans form. This indicates that isomerization of beta-carotene takes place during or after cyclization. PMID- 10677452 TI - Metabolic dysfunction and unabated respiration precede the loss of membrane integrity during dehydration of germinating radicles. AB - This study shows that dehydration induces imbalanced metabolism before loss of membrane integrity in desiccation-sensitive germinated radicles. Using a photoacoustic detection system, responses of CO(2) emission and fermentation to drying were analyzed non-invasively in desiccation-tolerant and -intolerant radicles of cucumber (Cucumis sativa) and pea (Pisum sativum). Survival after drying and a membrane integrity assay showed that desiccation tolerance was present during early imbibition and lost in germinated radicles. However, tolerance could be re-induced in germinated cucumber radicles by incubation in polyethylene glycol before drying. Tolerant and polyethylene glycol (PEG)-induced tolerant radicles exhibited a much-reduced CO(2) production before dehydration compared with desiccation-sensitive radicles. This difference was maintained during dehydration. In desiccation-sensitive tissues, dehydration induced an increase in the emission of acetaldehyde and ethanol that peaked well before the loss of membrane integrity. Acetaldehyde emission from sensitive radicles was significantly reduced when dehydration occurred in 50% O(2) instead of air. Acetaldehyde/ethanol were not detected in dehydrating tolerant radicles of either species or in polyethylene glycol-induced tolerant cucumber radicles. Thus, a balance between down-regulation of metabolism during drying and O(2) availability appears to be associated with desiccation tolerance. Using Fourier transform infrared spectroscopy, acetaldehyde was found to disturb the phase behavior of phospholipid vesicles, suggesting that the products resulting from imbalanced metabolism in seeds may aggravate membrane damage induced by dehydration. PMID- 10677454 TI - The electronic Plant Gene Register. PMID- 10677455 TI - The past climate change heats up. PMID- 10677456 TI - Of bears, conservation genetics, and the value of time travel. PMID- 10677457 TI - Closing the ring: links between SMC proteins and chromosome partitioning, condensation, and supercoiling. PMID- 10677458 TI - More surprises in translation: initiation without the initiator tRNA. PMID- 10677459 TI - Computation with biomolecules. PMID- 10677460 TI - Ice-core evidence of abrupt climate changes. AB - Ice-core records show that climate changes in the past have been large, rapid, and synchronous over broad areas extending into low latitudes, with less variability over historical times. These ice-core records come from high mountain glaciers and the polar regions, including small ice caps and the large ice sheets of Greenland and Antarctica. PMID- 10677461 TI - Nonglacial rapid climate events: past and future. AB - The paleoclimate record makes it clear that rapid climate shifts of the 20th century are only a subset of possible climate system behavior that might occur in the absence of glacial conditions, and that climatic surprises could be a challenge for society even in the absence of significant greenhouse warming. PMID- 10677462 TI - Was a change in thermohaline circulation responsible for the Little Ice Age? PMID- 10677463 TI - Detecting holocene changes in thermohaline circulation. AB - Throughout the last glacial cycle, reorganizations of deep ocean water masses were coincident with rapid millennial-scale changes in climate. Climate changes have been less severe during the present interglacial, but evidence for concurrent deep ocean circulation change is ambiguous. PMID- 10677464 TI - Abrupt climate change and thermohaline circulation: mechanisms and predictability. AB - The ocean's thermohaline circulation has long been recognized as potentially unstable and has consequently been invoked as a potential cause of abrupt climate change on all timescales of decades and longer. However, fundamental aspects of thermohaline circulation changes remain poorly understood. PMID- 10677465 TI - As climate changes, so do glaciers. AB - Understanding abrupt climate changes requires detailed spatial/temporal records of such changes, and to make these records, we need rapidly responding, geographically widespread climate trackers. Glacial systems are such trackers, and recent additions to the stratigraphic record show overall synchronous response of glacial systems to climate change reflecting global atmosphere conditions. PMID- 10677466 TI - Climate change and the tropical Pacific: the sleeping dragon wakes. PMID- 10677467 TI - Sensitivity and rapidity of vegetational response to abrupt climate change. AB - Rapid climate change characterizes numerous terrestrial sediment records during and since the last glaciation. Vegetational response is best expressed in terrestrial records near ecotones, where sensitivity to climate change is greatest, and response times are as short as decades. PMID- 10677468 TI - Abrupt climate change in the computer: is it real? AB - Models suggest that dramatic changes in the ocean circulation are responsible for abrupt climate changes during the last ice age and may possibly alter the relative climate stability of the last 10,000 years. PMID- 10677469 TI - Very intense pulse in the groundwater flow in fissurized-porous stratum. AB - An asymptotic solution is obtained corresponding to a very intense pulse: a sudden strong increase and fast subsequent decrease of the water level at the boundary of semi-infinite fissurized-porous stratum. This flow is of practical interest: it gives a model of a groundwater flow after a high water period or after a failure of a dam around a collector of liquid waste. It is demonstrated that the fissures have a dramatic influence on the groundwater flow, increasing the penetration depth and speed of fluid penetration into the stratum. A characteristic property of the flow in fissurized-porous stratum is the rapid breakthrough of the fluid at the first stage deeply into the stratum via a system of cracks, feeding of porous blocks by the fluid in cracks, and at a later stage feeding of advancing fluid flow in fissures by the fluid, accumulated in porous blocks. PMID- 10677470 TI - Archimedean solids: transition metal mediated rational self-assembly of supramolecular-truncated tetrahedra. AB - A family of nanoscale-sized supramolecular cage compounds with a polyhedral framework is prepared by self-assembly from tritopic building blocks and rectangular corner units via noncovalent coordination interactions. These highly symmetrical cage compounds are described as face-directed, self-assembled truncated tetrahedra with T(d) symmetry. PMID- 10677471 TI - Molecular computation: RNA solutions to chess problems. AB - We have expanded the field of "DNA computers" to RNA and present a general approach for the solution of satisfiability problems. As an example, we consider a variant of the "Knight problem," which asks generally what configurations of knights can one place on an n x n chess board such that no knight is attacking any other knight on the board. Using specific ribonuclease digestion to manipulate strands of a 10-bit binary RNA library, we developed a molecular algorithm and applied it to a 3 x 3 chessboard as a 9-bit instance of this problem. Here, the nine spaces on the board correspond to nine "bits" or placeholders in a combinatorial RNA library. We recovered a set of "winning" molecules that describe solutions to this problem. PMID- 10677472 TI - Two terrestrial records of rapid climatic change during the glacial-Holocene transition (14,000- 9,000 calendar years B.P.) from Europe. AB - Two independent multidisciplinary studies of climatic change during the glacial Holocene transition (ca. 14,000-9,000 calendar yr B.P.) from Norway and Switzerland have assessed organism responses to the rapid climatic changes and made quantitative temperature reconstructions with modern calibration data sets (transfer functions). Chronology at Krakenes, western Norway, was derived from calibration of a high-resolution series of 14C dates. Chronologies at Gerzensee and Leysin, Switzerland, were derived by comparison of delta18O in lake carbonates with the delta18O record from the Greenland Ice Core Project. Both studies demonstrate the sensitivity of terrestrial and aquatic organisms to rapid temperature changes and their value for quantitative reconstruction of the magnitudes and rates of the climatic changes. The rates in these two terrestrial records are comparable to those in Greenland ice cores, but the actual temperatures inferred apply to the terrestrial environments of the two regions. PMID- 10677473 TI - Paleoproterozoic snowball earth: extreme climatic and geochemical global change and its biological consequences. AB - Geological, geophysical, and geochemical data support a theory that Earth experienced several intervals of intense, global glaciation ("snowball Earth" conditions) during Precambrian time. This snowball model predicts that postglacial, greenhouse-induced warming would lead to the deposition of banded iron formations and cap carbonates. Although global glaciation would have drastically curtailed biological productivity, melting of the oceanic ice would also have induced a cyanobacterial bloom, leading to an oxygen spike in the euphotic zone and to the oxidative precipitation of iron and manganese. A Paleoproterozoic snowball Earth at 2.4 Giga-annum before present (Ga) immediately precedes the Kalahari Manganese Field in southern Africa, suggesting that this rapid and massive change in global climate was responsible for its deposition. As large quantities of O(2) are needed to precipitate this Mn, photosystem II and oxygen radical protection mechanisms must have evolved before 2.4 Ga. This geochemical event may have triggered a compensatory evolutionary branching in the Fe/Mn superoxide dismutase enzyme, providing a Paleoproterozoic calibration point for studies of molecular evolution. PMID- 10677474 TI - Twentieth century climate change: evidence from small glaciers. AB - The relation between changes in modern glaciers, not including the ice sheets of Greenland and Antarctica, and their climatic environment is investigated to shed light on paleoglacier evidence of past climate change and for projecting the effects of future climate warming on cold regions of the world. Loss of glacier volume has been more or less continuous since the 19th century, but it is not a simple adjustment to the end of an "anomalous" Little Ice Age. We address the 1961-1997 period, which provides the most observational data on volume changes. These data show trends that are highly variable with time as well as within and between regions; trends in the Arctic are consistent with global averages but are quantitatively smaller. The averaged annual volume loss is 147 mm.yr(-1) in water equivalent, totaling 3.7 x 10(3) km(3) over 37 yr. The time series shows a shift during the mid-1970s, followed by more rapid loss of ice volume and further acceleration in the last decade; this is consistent with climatologic data. Perhaps most significant is an increase in annual accumulation along with an increase in melting; these produce a marked increase in the annual turnover or amplitude. The rise in air temperature suggested by the temperature sensitivities of glaciers in cold regions is somewhat greater than the global average temperature rise derived largely from low altitude gauges, and the warming is accelerating. PMID- 10677475 TI - Can ozone depletion and global warming interact to produce rapid climate change? AB - The atmosphere displays modes of variability whose structures exhibit a strong longitudinally symmetric (annular) component that extends from the surface to the stratosphere in middle and high latitudes of both hemispheres. In the past 30 years, these modes have exhibited trends that seem larger than their natural background variability, and may be related to human influences on stratospheric ozone and/or atmospheric greenhouse gas concentrations. The pattern of climate trends during the past few decades is marked by rapid cooling and ozone depletion in the polar lower stratosphere of both hemispheres, coupled with an increasing strength of the wintertime westerly polar vortex and a poleward shift of the westerly wind belt at the earth's surface. Annular modes of variability are fundamentally a result of internal dynamical feedbacks within the climate system, and as such can show a large response to rather modest external forcing. The dynamics and thermodynamics of these modes are such that strong synergistic interactions between stratospheric ozone depletion and greenhouse warming are possible. These interactions may be responsible for the pronounced changes in tropospheric and stratospheric climate observed during the past few decades. If these trends continue, they could have important implications for the climate of the 21st century. PMID- 10677476 TI - Twist fields, the elliptic genus, and hidden symmetry. AB - We combine infinite dimensional analysis (in particular a priori estimates and twist positivity) with classical geometric structures, supersymmetry, and noncommutative geometry. We establish the existence of a family of examples of two-dimensional, twist quantum fields. We evaluate the elliptic genus in these examples. We demonstrate a hidden SL(2,Z) symmetry of the elliptic genus, as suggested by Witten. PMID- 10677477 TI - The multivariate L1-median and associated data depth. AB - This paper gives three related results: (i) a new, simple, fast, monotonically converging algorithm for deriving the L1-median of a data cloud in Rd, a problem that can be traced to Fermat and has fascinated applied mathematicians for over three centuries; (ii) a new general definition for depth functions, as functions of multivariate medians, so that different definitions of medians will, correspondingly, give rise to different dept functions; and (iii) a simple closed form formula of the L1-depth function for a given data cloud in Rd. PMID- 10677478 TI - Evolutionary conservation of apoptosis mechanisms: lepidopteran and baculoviral inhibitor of apoptosis proteins are inhibitors of mammalian caspase-9. AB - We cloned a new inhibitor of apoptosis protein (IAP) homolog, SfIAP, from Spodoptera frugiperda Sf-21 cells, a host of insect baculoviruses. SfIAP contains two baculovirus IAP repeat domains followed by a RING domain. SfIAP has striking amino acid sequence similarity with baculoviral IAPs, CpIAP and OpIAP, suggesting that baculoviral IAPs may be host-derived genes. SfIAP and baculoviral CpIAP inhibit Bax but not Fas-induced apoptosis in human cells. Their apoptosis suppressing activity in mammalian cells requires both baculovirus IAP repeat and RING domains. Further biochemical data suggest that SfIAP and CpIAP are specific inhibitors of mammalian caspase-9, the pinnacle caspase in the mitochondria/cytochrome c pathway for apoptosis, but are not inhibitors of downstream caspase-3 and caspase-7. Thus the mechanisms by which insect and baculoviral IAPs suppress apoptosis may involve inhibition of an insect caspase-9 homologue. Peptides representing the IAP-binding domain of the Drosophila cell death protein Grim abrogated human caspase suppression by SfIAP and CpIAP, implying evolutionary conservation of the functions of IAPs and their inhibitors. PMID- 10677479 TI - Synergy in a medicinal plant: antimicrobial action of berberine potentiated by 5' methoxyhydnocarpin, a multidrug pump inhibitor. AB - Multidrug resistance pumps (MDRs) protect microbial cells from both synthetic and natural antimicrobials. Amphipathic cations are preferred substrates of MDRs. Berberine alkaloids, which are cationic antimicrobials produced by a variety of plants, are readily extruded by MDRs. Several Berberis medicinal plants producing berberine were found also to synthesize an inhibitor of the NorA MDR pump of a human pathogen Staphylococcus aureus. The inhibitor was identified as 5' methoxyhydnocarpin (5'-MHC), previously reported as a minor component of chaulmoogra oil, a traditional therapy for leprosy. 5'-MHC is an amphipathic weak acid and is distinctly different from the cationic substrates of NorA. 5'-MHC had no antimicrobial activity alone but strongly potentiated the action of berberine and other NorA substrates against S. aureus. MDR-dependent efflux of ethidium bromide and berberine from S. aureus cells was completely inhibited by 5'-MHC. The level of accumulation of berberine in the cells was increased strongly in the presence of 5'-MHC, indicating that this plant compound effectively disabled the bacterial resistance mechanism against the berberine antimicrobial. PMID- 10677480 TI - Climate and infectious disease: use of remote sensing for detection of Vibrio cholerae by indirect measurement. AB - It has long been known that cholera outbreaks can be initiated when Vibrio cholerae, the bacterium that causes cholera, is present in drinking water in sufficient numbers to constitute an infective dose, if ingested by humans. Outbreaks associated with drinking or bathing in unpurified river or brackish water may directly or indirectly depend on such conditions as water temperature, nutrient concentration, and plankton production that may be favorable for growth and reproduction of the bacterium. Although these environmental parameters have routinely been measured by using water samples collected aboard research ships, the available data sets are sparse and infrequent. Furthermore, shipboard data acquisition is both expensive and time-consuming. Interpolation to regional scales can also be problematic. Although the bacterium, V. cholerae, cannot be sensed directly, remotely sensed data can be used to infer its presence. In the study reported here, satellite data were used to monitor the timing and spread of cholera. Public domain remote sensing data for the Bay of Bengal were compared directly with cholera case data collected in Bangladesh from 1992-1995. The remote sensing data included sea surface temperature and sea surface height. It was discovered that sea surface temperature shows an annual cycle similar to the cholera case data. Sea surface height may be an indicator of incursion of plankton-laden water inland, e.g., tidal rivers, because it was also found to be correlated with cholera outbreaks. The extensive studies accomplished during the past 25 years, confirming the hypothesis that V. cholerae is autochthonous to the aquatic environment and is a commensal of zooplankton, i.e., copepods, when combined with the findings of the satellite data analyses, provide strong evidence that cholera epidemics are climate-linked. PMID- 10677481 TI - The subcellular localization of acetyl-CoA carboxylase 2. AB - Animals, including humans, express two isoforms of acetyl-CoA carboxylase (EC ), ACC1 (M(r) = 265 kDa) and ACC2 (M(r) = 280 kDa). The predicted amino acid sequence of ACC2 contains an additional 136 aa relative to ACC1, 114 of which constitute the unique N-terminal sequence of ACC2. The hydropathic profiles of the two ACC isoforms generally are comparable, except for the unique N-terminal sequence in ACC2. The sequence of amino acid residues 1-20 of ACC2 is highly hydrophobic, suggesting that it is a leader sequence that targets ACC2 for insertion into membranes. The subcellular localization of ACC2 in mammalian cells was determined by performing immunofluorescence microscopic analysis using affinity-purified anti-ACC2-specific antibodies and transient expression of the green fluorescent protein fused to the C terminus of the N-terminal sequences of ACC1 and ACC2. These analyses demonstrated that ACC1 is a cytosolic protein and that ACC2 was associated with the mitochondria, a finding that was confirmed further by the immunocolocalization of a known human mitochondria-specific protein and the carnitine palmitoyltransferase 1. Based on analyses of the fusion proteins of ACC-green fluorescent protein, we concluded that the N-terminal sequences of ACC2 are responsible for mitochondrial targeting of ACC2. The association of ACC2 with the mitochondria is consistent with the hypothesis that ACC2 is involved in the regulation of mitochondrial fatty acid oxidation through the inhibition of carnitine palmitoyltransferase 1 by its product malonyl-CoA. PMID- 10677482 TI - A structural basis for integrin activation by the cytoplasmic tail of the alpha IIb-subunit. AB - A key step in the activation of heterodimeric integrin adhesion receptors is the transmission of an agonist-induced cellular signal from the short alpha- and/or beta-cytoplasmic tails to the extracellular domains of the receptor. The structural details of how the cytoplasmic tails mediate such an inside-out signaling process remain unclear. We report herein the NMR structures of a membrane-anchored cytoplasmic tail of the alpha(IIb)-subunit and of a mutant alpha(IIb)-cytoplasmic tail that renders platelet integrin alpha(IIb)beta(3) constitutively active. The structure of the wild-type alpha(IIb)-cytoplasmic tail reveals a "closed" conformation where the highly conserved N-terminal membrane proximal region forms an alpha-helix followed by a turn, and the acidic C terminal loop interacts with the N-terminal helix. The structure of the active mutant is significantly different, having an "open" conformation where the interactions between the N-terminal helix and C-terminal region are abolished. Consistent with these structural differences, the two peptides differ in function: the wild-type peptide suppressed alpha(IIb)beta(3) activation, whereas the mutant peptide did not. These results provide an atomic explanation for extensive biochemical/mutational data and support a conformation-based "on/off switch" model for integrin activation. PMID- 10677483 TI - Human aspartic protease memapsin 2 cleaves the beta-secretase site of beta amyloid precursor protein. AB - The cDNAs of two new human membrane-associated aspartic proteases, memapsin 1 and memapsin 2, have been cloned and sequenced. The deduced amino acid sequences show that each contains the typical pre, pro, and aspartic protease regions, but each also has a C-terminal extension of over 80 residues, which includes a single transmembrane domain and a C-terminal cytosolic domain. Memapsin 2 mRNA is abundant in human brain. The protease domain of memapsin 2 cDNA was expressed in Escherichia coli and was purified. Recombinant memapsin 2 specifically hydrolyzed peptides derived from the beta-secretase site of both the wild-type and Swedish mutant beta-amyloid precursor protein (APP) with over 60-fold increase of catalytic efficiency for the latter. Expression of APP and memapsin 2 in HeLa cells showed that memapsin 2 cleaved the beta-secretase site of APP intracellularly. These and other results suggest that memapsin 2 fits all of the criteria of beta-secretase, which catalyzes the rate-limiting step of the in vivo production of the beta-amyloid (Abeta) peptide leading to the progression of Alzheimer's disease. Recombinant memapsin 2 also cleaved a peptide derived from the processing site of presenilin 1, albeit with poor kinetic efficiency. Alignment of cleavage site sequences of peptides indicates that the specificity of memapsin 2 resides mainly at the S(1)' subsite, which prefers small side chains such as Ala, Ser, and Asp. PMID- 10677484 TI - Consequences of placing an intramolecular crosslink in myosin S1. AB - This paper describes the placement of a crosslinking agent (dibromobimane) between two thiols (Cys-522 and Cys-707) of a fragment, "S1," of the motor protein, myosin. It turns out that fastening the first anchor of the crosslinker is easy and rapid, but fastening the second anchor (Cys-522) is very temperature dependent, taking 30 min at room temperature but about a week on ice. Moreover, crystallography taken at 4 degrees C would seem to predict that the linkage is impossible, because the span of the crosslinking agent is much less than the interthiol distance. The simplest resolution of this seeming paradox is that structural fluctuations of the protein render the linkage increasingly likely as the temperature increases. Also, measurements of the affinity of MgADP for the protein, as well as the magnetic resonance of the P-atoms of the ADP once emplaced, suggest that binding the first reagent anchor to Cys-707 initiates an influence that travels to the rather distant ADP-binding site, and it is speculated what this "path of influence" might be. PMID- 10677485 TI - Three-dimensional modeling of and ligand docking to vitamin D receptor ligand binding domain. AB - The ligand binding domain of the human vitamin D receptor (VDR) was modeled based on the crystal structure of the retinoic acid receptor. The ligand binding pocket of our VDR model is spacious at the helix 11 site and confined at the beta-turn site. The ligand 1alpha, 25-dihydroxyvitamin D(3) was assumed to be anchored in the ligand binding pocket with its side chain heading to helix 11 (site 2) and the A-ring toward the beta-turn (site 1). Three residues forming hydrogen bonds with the functionally important 1alpha- and 25-hydroxyl groups of 1alpha,25 dihydroxyvitamin D(3) were identified and confirmed by mutational analysis: the 1alpha-hydroxyl group is forming pincer-type hydrogen bonds with S237 and R274 and the 25-hydroxyl group is interacting with H397. Docking potential for various ligands to the VDR model was examined, and the results are in good agreement with our previous three-dimensional structure-function theory. PMID- 10677486 TI - Self-assembly of large, ordered lamellae from non-bilayer lipids and integral membrane proteins in vitro. AB - In many biological membranes, the major lipids are "non-bilayer lipids," which in purified form cannot be arranged in a lamellar structure. The structural and functional roles of these lipids are poorly understood. This work demonstrates that the in vitro association of the two main components of a membrane, the non bilayer lipid monogalactosyldiacylglycerol (MGDG) and the chlorophyll-a/b light harvesting antenna protein of photosystem II (LHCII) of pea thylakoids, leads to the formation of large, ordered lamellar structures: (i) thin-section electron microscopy and circular dichroism spectroscopy reveal that the addition of MGDG induces the transformation of isolated, disordered macroaggregates of LHCII into stacked lamellar aggregates with a long-range chiral order of the complexes; (ii) small-angle x-ray scattering discloses that LHCII perturbs the structure of the pure lipid and destroys the inverted hexagonal phase; and (iii) an analysis of electron micrographs of negatively stained 2D crystals indicates that in MGDG LHCII the complexes are found in an ordered macroarray. It is proposed that, by limiting the space available for MGDG in the macroaggregate, LHCII inhibits formation of the inverted hexagonal phase of lipids; in thylakoids, a spatial limitation is likely to be imposed by the high concentration of membrane associated proteins. PMID- 10677487 TI - Folding and activity of circularly permuted forms of a polytopic membrane protein. AB - The transmembrane subunit of the Glc transporter (IICB(Glc)), which mediates uptake and concomitant phosphorylation of glucose, spans the membrane eight times. Variants of IICB(Glc) with the native N and C termini joined and new N and C termini in the periplasmic and cytoplasmic surface loops were expressed in Escherichia coli. In vivo transport/in vitro phosphotransferase activities of the circularly permuted variants with the termini in the periplasmic loops 1 to 4 were 35/58, 32/37, 0/3, and 0/0% of wild type, respectively. The activities of the variants with the termini in the cytoplasmic loops 1 to 3 were 0/25, 0/4 and 24/70, respectively. Fusion of alkaline phosphatase to the periplasmic C termini stabilized membrane integration and increased uptake and/or phosphorylation activities. These results suggest that internal signal anchor and stop transfer sequences can function as N-terminal signal sequences in a circularly permuted alpha-helical bundle protein and that the orientation of transmembrane segments is determined by the amino acid sequence and not by the sequential appearance during translation. Of the four IICB(Glc) variants with new termini in periplasmic loops, only the one with the discontinuity in loop 4 is inactive. The sequences of loop 4 and of the adjacent TM7 and TM8 are conserved in all phosphoenolpyruvate-dependent carbohydrate:phosphotransferase system transporters of the glucose family. PMID- 10677488 TI - A regulator of G protein signaling interaction surface linked to effector specificity. AB - Proteins of the regulator of G protein signaling (RGS) family accelerate GTP hydrolysis by the alpha subunits (G(alpha)) of G proteins, leading to rapid recovery of signaling cascades. Many different RGS proteins can accelerate GTP hydrolysis by an individual G(alpha), and GTP hydrolysis rates of different G(alpha)s can be enhanced by the same RGS protein. Consequently, the mechanisms for specificity in RGS regulation and the residues involved remain unclear. Using the evolutionary trace (ET) method, we have identified a cluster of residues in the RGS domain that includes the RGS-G(alpha) binding interface and extends to include additional functionally important residues on the surface. One of these is within helix alpha3, two are in alpha5, and three are in the loop connecting alpha5 and alpha6. A cluster of surface residues on G(alpha) previously identified by ET, and composed predominantly of residues from the switch III region and helix alpha3, is spatially contiguous with the ET-identified residues in the RGS domain. This cluster includes residues proposed to interact with the gamma subunit of G(talpha)'s effector, cGMP phosphodiesterase (PDEgamma). The proximity of these clusters suggests that they form part of an interface between the effector and the RGS-G(alpha) complex. Sequence variations in these residues correlate with PDEgamma effects on GTPase acceleration. Because ET identifies residues important for all members of a protein family, these residues likely form a general site for regulation of G protein-coupled signaling cascades, possibly by means of effector interactions. PMID- 10677489 TI - Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors. AB - Acting through a number of distinct pathways, many G protein-coupled receptors (GPCRs) activate the extracellular signal-regulated kinase (ERK)/mitogen activated protein kinase (MAPK) cascade. Recently, it has been shown that in some cases, clathrin-mediated endocytosis is required for GPCR activation of the ERK/MAPK cascade, whereas in others it is not. Accordingly, we compared ERK activation mediated by a GPCR that does not undergo agonist-stimulated endocytosis, the alpha(2A) adrenergic receptor (alpha(2A) AR), with ERK activation mediated by the beta(2) adrenergic receptor (beta(2) AR), which is endocytosed. Surprisingly, we found that in COS-7 cells, ERK activation by the alpha(2A) AR, like that mediated by both the beta(2) AR and the epidermal growth factor receptor (EGFR), is sensitive to mechanistically distinct inhibitors of clathrin-mediated endocytosis, including monodansylcadaverine, a mutant dynamin I, and a mutant beta-arrestin 1. Moreover, we determined that, as has been shown for many other GPCRs, both alpha(2A) and beta(2) AR-mediated ERK activation involves transactivation of the EGFR. Using confocal immunofluorescence microscopy, we found that stimulation of the beta(2) AR, the alpha(2A) AR, or the EGFR each results in internalization of a green fluorescent protein-tagged EGFR. Although beta(2) AR stimulation leads to redistribution of both the beta(2) AR and EGFR, activation of the alpha(2A) AR leads to redistribution of the EGFR but the alpha(2A) AR remains on the plasma membrane. These findings separate GPCR endocytosis from the requirement for clathrin-mediated endocytosis in EGFR transactivation-mediated ERK activation and suggest that it is the receptor tyrosine kinase or another downstream effector that must engage the endocytic machinery. PMID- 10677490 TI - Host factor Hfq of Escherichia coli stimulates elongation of poly(A) tails by poly(A) polymerase I. AB - Current evidence suggests that the length of poly(A) tails of bacterial mRNAs result from a competition between poly(A) polymerase and exoribonucleases that attack the 3' ends of RNAs. Here, we show that host factor Hfq is also involved in poly(A) tail metabolism. Inactivation of the hfq gene reduces the length of poly(A) tails synthesized at the 3' end of the rpsO mRNA by poly(A) polymerase I in vivo. In vitro, Hfq stimulates synthesis of long tails by poly(A) polymerase I. The strong binding of Hfq to oligoadenylated RNA probably explains why it stimulates elongation of primers that already harbor tails of 20-35 A. Polyadenylation becomes processive in the presence of Hfq. The similar properties of Hfq and the PABPII poly(A) binding protein, which stimulates poly(A) tail elongation in mammals, indicates that similar mechanisms control poly(A) tail synthesis in prokaryotes and eukaryotes. PMID- 10677491 TI - Allosteric inhibitors of inducible nitric oxide synthase dimerization discovered via combinatorial chemistry. AB - Potent and selective inhibitors of inducible nitric oxide synthase (iNOS) (EC ) were identified in an encoded combinatorial chemical library that blocked human iNOS dimerization, and thereby NO production. In a cell-based iNOS assay (A-172 astrocytoma cells) the inhibitors had low-nanomolar IC(50) values and thus were >1,000-fold more potent than the substrate-based direct iNOS inhibitors 1400W and N-methyl-l-arginine. Biochemical studies confirmed that inhibitors caused accumulation of iNOS monomers in mouse macrophage RAW 264.7 cells. High affinity (K(d) approximately 3 nM) of inhibitors for isolated iNOS monomers was confirmed by using a radioligand binding assay. Inhibitors were >1,000-fold selective for iNOS versus endothelial NOS dimerization in a cell-based assay. The crystal structure of inhibitor bound to the monomeric iNOS oxygenase domain revealed inhibitor-heme coordination and substantial perturbation of the substrate binding site and the dimerization interface, indicating that this small molecule acts by allosterically disrupting protein-protein interactions at the dimer interface. These results provide a mechanism-based approach to highly selective iNOS inhibition. Inhibitors were active in vivo, with ED(50) values of <2 mg/kg in a rat model of endotoxin-induced systemic iNOS induction. Thus, this class of dimerization inhibitors has broad therapeutic potential in iNOS-mediated pathologies. PMID- 10677492 TI - Studies on the role of the hydrophobic domain of Ost4p in interactions with other subunits of yeast oligosaccharyl transferase. AB - In the yeast, Saccharomyces cerevisiae, oligosaccharyl transferase (OT), which catalyzes the transfer of dolichol-linked oligosaccharide chains to nascent polypeptides in the endoplasmic reticulum, consists of nine nonidentical membrane protein subunits. Genetic and biochemical evidence indicated these nine proteins exist in three subcomplexes. Three of the OT subunits (Ost4p, Ost3p, and Stt3p) have been proposed to exist in one subcomplex. To investigate the interaction of these three membrane proteins, initially we carried out a mutational analysis of Ost4p, which is an extraordinarily small membrane protein containing only 36 amino acid residues. This analysis indicated that when single amino acid residues in a region close to the luminal face of the putative transmembrane domain of Ost4p were changed into an ionizable amino acid such as Lys or Asp, growth at 37 degrees C and OT activity measured in vitro were impaired. In addition, using immunoprecipitation techniques and Western blot analysis, we found that with these mutations the interaction between Ost4p, Ost3p, and Stt3p was disrupted. Introduction of Lys or Asp residues at other positions in the putative transmembrane domain or at the N or C terminus of Ost4p had no effect on disrupting subunit interactions or impairing the activity of OT. These findings suggest that a localized region of the putative transmembrane domain of Ost4p mediates in stabilization of the interaction with the two other OT subunits (Ost3p and Stt3p) in a subcomplex in the endoplasmic reticulum membrane. PMID- 10677493 TI - Coupling between protein folding and allostery in the GroE chaperonin system. AB - GroEL is an allosteric protein that facilitates protein folding in an ATP dependent manner. Herein, the relationship between cooperative ATP binding by GroEL and the kinetics of GroE-assisted folding of two substrates with different GroES dependence, mouse dihydrofolate reductase (mDHFR) and mitochondrial malate dehydrogenase, is examined by using cooperativity mutants of GroEL. Strong intra ring positive cooperativity in ATP binding by GroEL decreases the rate of GroEL assisted mDHFR folding owing to a slow rate of the ATP-induced transition from the protein-acceptor state to the protein-release state. Inter-ring negative cooperativity in ATP binding by GroEL is found to affect the kinetic partitioning of mDHFR, but not of mitochondrial malate dehydrogenase, between folding in solution and folding in the cavity underneath GroES. Our results show that protein folding by this "two-stroke motor" is coupled to cooperative ATP binding. PMID- 10677494 TI - Transition-state structure as a unifying basis in protein-folding mechanisms: contact order, chain topology, stability, and the extended nucleus mechanism. AB - I attempt to reconcile apparently conflicting factors and mechanisms that have been proposed to determine the rate constant for two-state folding of small proteins, on the basis of general features of the structures of transition states. Phi-Value analysis implies a transition state for folding that resembles an expanded and distorted native structure, which is built around an extended nucleus. The nucleus is composed predominantly of elements of partly or well formed native secondary structure that are stabilized by local and long-range tertiary interactions. These long-range interactions give rise to connecting loops, frequently containing the native loops that are poorly structured. I derive an equation that relates differences in the contact order of a protein to changes in the length of linking loops, which, in turn, is directly related to the unfavorable free energy of the loops in the transition state. Kinetic data on loop extension mutants of CI2 and alpha-spectrin SH3 domain fit the equation qualitatively. The rate of folding depends primarily on the interactions that directly stabilize the nucleus, especially those in native-like secondary structure and those resulting from the entropy loss from the connecting loops, which vary with contact order. This partitioning of energy accounts for the success of some algorithms that predict folding rates, because they use these principles either explicitly or implicitly. The extended nucleus model thus unifies the observations of rate depending on both stability and topology. PMID- 10677495 TI - A double-hexamer archaeal minichromosome maintenance protein is an ATP-dependent DNA helicase. AB - The minichromosome maintenance (MCM) proteins are essential for DNA replication in eukaryotes. Thus far, all eukaryotes have been shown to contain six highly related MCMs that apparently function together in DNA replication. Sequencing of the entire genome of the thermophilic archaeon Methanobacterium thermoautotrophicum has allowed us to identify only a single MCM-like gene (ORF Mt1770). This gene is most similar to MCM4 in eukaryotic cells. Here we have expressed and purified the M. thermoautotrophicum MCM protein. The purified protein forms a complex that has a molecular mass of approximately 850 kDa, consistent with formation of a double hexamer. The protein has an ATP-independent DNA-binding activity, a DNA-stimulated ATPase activity that discriminates between single- and double-stranded DNA, and a strand-displacement (helicase) activity that can unwind up to 500 base pairs. The 3' to 5' helicase activity requires both ATP hydrolysis and a functional nucleotide-binding site. Moreover, the double hexamer form is the active helicase. It is therefore likely that an MCM complex acts as the replicative DNA helicase in eukaryotes and archaea. The simplified replication machinery in archaea may provide a simplified model for assembly of the machinery required for initiation of eukaryotic DNA replication. PMID- 10677496 TI - A 9-nt segment of a cellular mRNA can function as an internal ribosome entry site (IRES) and when present in linked multiple copies greatly enhances IRES activity. AB - This study addresses the properties of a newly identified internal ribosome entry site (IRES) contained within the mRNA of the homeodomain protein Gtx. Sequential deletions of the 5' untranslated region (UTR) from either end did not define distinct IRES boundaries; when five nonoverlapping UTR fragments were tested, four had IRES activity. These observations are consistent with other cellular IRES analyses suggesting that some cellular IRESes are composed of segments (IRES modules) that independently and combinatorially contribute to overall IRES activity. We characterize a 9-nt IRES module from the Gtx 5' UTR that is 100% complementary to the 18S rRNA at nucleotides 1132-1124. In previous work, we demonstrated that this mRNA segment could be crosslinked to its complement within intact 40S subunits. Here we show that increasing the number of copies of this IRES module in the intercistronic region of a dicistronic mRNA strongly enhances IRES activity in various cell lines. Ten linked copies increased IRES activity up to 570-fold in Neuro 2a cells. This level of IRES activity is up to 63-fold greater than that obtained by using the well characterized encephalomyocarditis virus IRES when tested in the same assay system. When the number of nucleotides between two of the 9-nt Gtx IRES modules was increased, the synergy between them decreased. In light of these findings, we discuss possible mechanisms of ribosome recruitment by cellular mRNAs, address the proposed role of higher order RNA structures on cellular IRES activity, and suggest parallels between IRES modules and transcriptional enhancer elements. PMID- 10677497 TI - Determination of the binding sites of the proton transfer inhibitors Cd2+ and Zn2+ in bacterial reaction centers. AB - The reaction center (RC) from Rhodobacter sphaeroides couples light-driven electron transfer to protonation of a bound quinone acceptor molecule, Q(B), within the RC. The binding of Cd(2+) or Zn(2+) has been previously shown to inhibit the rate of reduction and protonation of Q(B). We report here on the metal binding site, determined by x-ray diffraction at 2.5-A resolution, obtained from RC crystals that were soaked in the presence of the metal. The structures were refined to R factors of 23% and 24% for the Cd(2+) and Zn(2+) complexes, respectively. Both metals bind to the same location, coordinating to Asp-H124, His-H126, and His-H128. The rate of electron transfer from Q(A)(-) to Q(B) was measured in the Cd(2+)-soaked crystal and found to be the same as in solution in the presence of Cd(2+). In addition to the changes in the kinetics, a structural effect of Cd(2+) on Glu-H173 was observed. This residue was well resolved in the x-ray structure-i.e., ordered-with Cd(2+) bound to the RC, in contrast to its disordered state in the absence of Cd(2+), which suggests that the mobility of Glu-H173 plays an important role in the rate of reduction of Q(B). The position of the Cd(2+) and Zn(2+) localizes the proton entry into the RC near Asp-H124, His-H126, and His-H128. Based on the location of the metal, likely pathways of proton transfer from the aqueous surface to Q(B) are proposed. PMID- 10677498 TI - Identification of the proton pathway in bacterial reaction centers: replacement of Asp-M17 and Asp-L210 with asn reduces the proton transfer rate in the presence of Cd2+. AB - The reaction center (RC) from Rhodobacter sphaeroides converts light into chemical energy through the reduction and protonation of a bound quinone molecule Q(B) (the secondary quinone electron acceptor). We investigated the proton transfer pathway by measuring the proton-coupled electron transfer, k(AB)((2)) [Q(A)Q(B) + H(+) --> Q(A)(Q(B)H)(-)] in native and mutant RCs in the absence and presence of Cd(2+). Previous work has shown that the binding of Cd(2+) decreases k(AB)((2)) in native RCs approximately 100-fold. The preceding paper shows that bound Cd(2+) binds to Asp-H124, His-H126, and His-H128. This region represents the entry point for protons. In this work we investigated the proton transfer pathway connecting the entry point with Q(B) by searching for mutations that greatly affect k(AB)((2)) ( greater, similar10-fold) in the presence of Cd(2+), where k(AB)((2)) is limited by the proton transfer rate (k(H)). Upon mutation of Asp-L210 or Asp-M17 to Asn, k(H) decreased from approximately 60 s(-1) to approximately 7 s(-1), which shows the important role that Asp-L210 and Asp-M17 play in the proton transfer chain. By comparing the rate of proton transfer in the mutants (k(H) approximately 7 s(-1)) with that in native RCs in the absence of Cd(2+) (k(H) >/= 10(4) s(-1)), we conclude that alternate proton transfer pathways, which have been postulated, are at least 10(3)-fold less effective. PMID- 10677499 TI - The golgi-associated COPI-coated buds and vesicles contain beta/gamma -actin. AB - It has been shown previously that the morphology and subcellular positioning of the Golgi complex is controlled by actin microfilaments. To further characterize the association between actin microfilaments and the Golgi complex, we have used the Clostridium botulinum toxins C2 and C3, which specifically inhibit actin polymerization and cause depolymerization of F-actin in intact cells by the ADP ribosylation of G-actin monomers and the Rho small GTP-binding protein, respectively. Normal rat kidney cells treated with C2 showed that disruption of the actin and the collapse of the Golgi complex occurred concomitantly. However, when cells were treated with C3, the actin disassembly was observed without any change in the organization of the Golgi complex. The absence of the involvement of Rho was further confirmed by the treatment with lysophosphatidic acid or microinjection with the constitutively activated form of RhoA, both of which induced the stress fiber formation without affecting the Golgi complex. Immunogold electron microscopy in normal rat kidney cells revealed that beta- and gamma-actin isoforms were found in Golgi-associated COPI-coated buds and vesicles. Taken together, the results suggest that the Rho signaling pathway does not directly regulate Golgi-associated actin microfilaments, and that beta- and gamma-actins might be involved in the formation and/or transport of Golgi-derived vesicular or tubular intermediates. PMID- 10677500 TI - RANK is the intrinsic hematopoietic cell surface receptor that controls osteoclastogenesis and regulation of bone mass and calcium metabolism. AB - We have generated RANK (receptor activator of NF-kappaB) nullizygous mice to determine the molecular genetic interactions between osteoprotegerin, osteoprotegerin ligand, and RANK during bone resorption and remodeling processes. RANK(-/-) mice lack osteoclasts and have a profound defect in bone resorption and remodeling and in the development of the cartilaginous growth plates of endochondral bone. The osteopetrosis observed in these mice can be reversed by transplantation of bone marrow from rag1(-/-) (recombinase activating gene 1) mice, indicating that RANK(-/-) mice have an intrinsic defect in osteoclast function. Calciotropic hormones and proresorptive cytokines that are known to induce bone resorption in mice and human were administered to RANK(-/-) mice without inducing hypercalcemia, although tumor necrosis factor alpha treatment leads to the rare appearance of osteoclast-like cells near the site of injection. Osteoclastogenesis can be initiated in RANK(-/-) mice by transfer of the RANK cDNA back into hematopoietic precursors, suggesting a means to critically evaluate RANK structural features required for bone resorption. Together these data indicate that RANK is the intrinsic cell surface determinant that mediates osteoprotegerin ligand effects on bone resorption and remodeling as well as the physiological and pathological effects of calciotropic hormones and proresorptive cytokines. PMID- 10677501 TI - A functional genetic screen identifies regions at the C-terminal tail and death domain of death-associated protein kinase that are critical for its proapoptotic activity. AB - Death-associated protein kinase (DAP-kinase) is a Ca(+2)/calmodulin-regulated serine/threonine kinase with a multidomain structure that participates in apoptosis induced by a variety of signals. To identify regions in this protein that are critical for its proapoptotic activity, we performed a genetic screen on the basis of functional selection of short DAP-kinase-derived fragments that could protect cells from apoptosis by acting in a dominant-negative manner. We expressed a library of randomly fragmented DAP-kinase cDNA in HeLa cells and treated these cells with IFN-gamma to induce apoptosis. Functional cDNA fragments were recovered from cells that survived the selection, and those in the sense orientation were examined further in a secondary screen for their ability to protect cells from DAP-kinase-dependent tumor necrosis factor-alpha-induced apoptosis. We isolated four biologically active peptides that mapped to the ankyrin repeats, the "linker" region, the death domain, and the C-terminal tail of DAP-kinase. Molecular modeling of the complete death domain provided a structural basis for the function of the death-domain-derived fragment by suggesting that the protective fragment constitutes a distinct substructure. The last fragment, spanning the C-terminal serine-rich tail, defined a new regulatory region. Ectopic expression of the tail peptide (17 amino acids) inhibited the function of DAP-kinase, whereas removal of this region from the complete protein caused enhancement of the killing activity, indicating that the C-terminal tail normally plays a negative regulatory role. Altogether, this unbiased screen highlighted functionally important regions in the protein and revealed an additional level of regulation of DAP-kinase apoptotic function that does not affect the catalytic activity. PMID- 10677502 TI - Survival function of ERK1/2 as IL-3-activated, staurosporine-resistant Bcl2 kinases. AB - Bcl2 phosphorylation at Ser-70 may be required for the full and potent suppression of apoptosis in IL-3-dependent myeloid cells and can result from agonist activation of mitochondrial protein kinase C (PKC). Paradoxically, expression of exogenous Bcl2 can protect parental cells from apoptosis induced by the potent PKC inhibitor, staurosporine (stauro). High concentrations of stauro of up to 1 microM only partially inhibit IL-3-stimulated Bcl2 phosphorylation but completely block PKC-mediated Bcl2 phosphorylation in vitro. These data indicate a role for a stauro-resistant Bcl2 kinase (SRK). We show that aurintricarboxylic acid (ATA), a nonpeptide activator of cellular MEK/mitogen-activated protein kinase (MAPK) kinase, can induce Ser-70 phosphorylation of Bcl2 and support survival of cells expressing wild-type but not the phosphorylation-incompetent S70A mutant Bcl2. A role for a MEK/MAPK as a responsible SRK was implicated because the highly specific MEK/MAPK inhibitor, PD98059, also can only partially inhibit IL-3-induced Bcl2 phosphorylation, whereas the combination of PD98059 and stauro completely blocks phosphorylation and synergistically enhances apoptosis. p44MAPK/extracellular signal-regulated kinase 1 (ERK1) and p42 MAPK/ERK2 are activated by IL-3, colocalize with mitochondrial Bcl2, and can directly phosphorylate Bcl2 on Ser-70 in a stauro-resistant manner both in vitro and in vivo. These findings suggest a role for the ERK1/2 kinases as SRKs. Thus, the SRKs can serve to functionally link the IL-3-stimulated proliferative and survival signaling pathways and, in a novel capacity, may explain how Bcl2 can suppress stauro-induced apoptosis. In addition, although the mechanism of regulation of Bcl2 by phosphorylation is not yet clear, our results indicate that phosphorylation may functionally stabilize the Bcl2-Bax heterodimerization. PMID- 10677503 TI - The catalytic subunit of DNA-dependent protein kinase selectively regulates p53 dependent apoptosis but not cell-cycle arrest. AB - DNA damage induced by ionizing radiation (IR) activates p53, leading to the regulation of downstream pathways that control cell-cycle progression and apoptosis. However, the mechanisms for the IR-induced p53 activation and the differential activation of pathways downstream of p53 are unclear. Here we provide evidence that the catalytic subunit of DNA-dependent protein kinase (DNA PKcs) serves as an upstream effector for p53 activation in response to IR, linking DNA damage to apoptosis. DNA-PKcs knockout (DNA-PKcs-/-) mice were exposed to whole-body IR, and the cell-cycle and apoptotic responses were examined in their thymuses. Our data show that IR induction of apoptosis and Bax expression, both mediated via p53, was significantly suppressed in the thymocytes of DNA-PKcs-/- mice. In contrast, IR-induced cell-cycle arrest and p21 expression were normal. Thus, DNA-PKcs deficiency selectively disrupts p53-dependent apoptosis but not cell-cycle arrest. We also confirmed previous findings that p21 induction was attenuated and cell-cycle arrest was defective in the thymoctyes of whole body-irradiated Atm-/- mice, but the apoptotic response was unperturbed. Taken together, our results support a model in which the upstream effectors DNA PKcs and Atm selectively activate p53 to differentially regulate cell-cycle and apoptotic responses. Whereas Atm selects for cell-cycle arrest but not apoptosis, DNA-PKcs selects for apoptosis but not cell-cycle arrest. PMID- 10677504 TI - Aggregation of huntingtin in yeast varies with the length of the polyglutamine expansion and the expression of chaperone proteins. AB - Huntington's disease (HD) is an inherited neurodegenerative disorder caused by polyglutamine (polyQ) expansions in the huntingtin (Ht) protein. A hallmark of HD is the proteolytic production of an N-terminal fragment of Ht, containing the polyQ repeat, that forms aggregates in the nucleus and cytoplasm of affected neurons. Proteins with longer polyQ repeats aggregate more rapidly and cause disease at an earlier age, but the mechanism of aggregation and its relationship to disease remain unclear. To provide a new, genetically tractable model system for the study of Ht, we engineered yeast cells to express an N-terminal fragment of Ht with different polyQ repeat lengths of 25, 47, 72, or 103 residues, fused to green fluorescent protein. The extent of aggregation varied with the length of the polyQ repeat: at the two extremes, most HtQ103 protein coalesced into a single large cytoplasmic aggregate, whereas HtQ25 exhibited no sign of aggregation. Mutations that inhibit the ubiquitin/proteasome pathway at three different steps had no effect on the aggregation of Ht fragments in yeast, suggesting that the ubiquitination of Ht previously noted in mammalian cells may not inherently be required for polyQ length-dependent aggregation. Changing the expression levels of a wide variety of chaperone proteins in yeast neither increased nor decreased Ht aggregation. However, Sis1, Hsp70, and Hsp104 overexpression modulated aggregation of HtQ72 and HtQ103 fragments. More dramatically, the deletion of Hsp104 virtually eliminated it. These observations establish yeast as a system for studying the causes and consequences of polyQ dependent Ht aggregation. PMID- 10677505 TI - Myeloblastin is a granulocyte colony-stimulating factor-responsive gene conferring factor-independent growth to hematopoietic cells. AB - Hematopoiesis depends on a pool of quiescent hematopoietic stem/progenitor cells. When exposed to specific cytokines, a portion of these cells enters the cell cycle to generate an amplified progeny. Myeloblastin (MBN) initially was described as involved in proliferation of human leukemia cells. The granulocyte colony-stimulating factor (G-CSF), which stimulates the proliferation of granulocytic precursors, up-regulates MBN expression. Here we show that constitutive overexpression of MBN confers factor-independent growth to murine bone marrow-derived Ba/F3/G-CSFR cells. Our results point to MBN as a G-CSF responsive gene critical to factor-independent growth and indicate that expression of the G-CSF receptor is a prerequisite to this process. A 91-bp MBN promoter region containing PU.1, C/EBP, and c-Myb binding sites is responsive to G-CSF treatment. Although PU.1, C/EBP, and c-Myb transcription factors all were critical for expression of MBN, its up-regulation by G-CSF was associated mainly with PU.1. These findings suggest that MBN is an important target of PU.1 and a key protease for factor-independent growth of hematopoietic cells. PMID- 10677506 TI - neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas. AB - In the mammalian pancreas, the endocrine cell types of the islets of Langerhans, including the alpha-, beta-, delta-, and pancreatic polypeptide cells as well as the exocrine cells, derive from foregut endodermal progenitors. Recent genetic studies have identified a network of transcription factors, including Pdx1, Isl1, Pax4, Pax6, NeuroD, Nkx2.2, and Hlxb9, regulating the development of islet cells at different stages, but the molecular mechanisms controlling the specification of pancreatic endocrine precursors remain unknown. neurogenin3 (ngn3) is a member of a family of basic helix-loop-helix transcription factors that is involved in the determination of neural precursor cells in the neuroectoderm. ngn3 is expressed in discrete regions of the nervous system and in scattered cells in the embryonic pancreas. We show herein that ngn3-positive cells coexpress neither insulin nor glucagon, suggesting that ngn3 marks early precursors of pancreatic endocrine cells. Mice lacking ngn3 function fail to generate any pancreatic endocrine cells and die postnatally from diabetes. Expression of Isl1, Pax4, Pax6, and NeuroD is lost, and endocrine precursors are lacking in the mutant pancreatic epithelium. Thus, ngn3 is required for the specification of a common precursor for the four pancreatic endocrine cell types. PMID- 10677507 TI - Efficient studies of long-distance Bmp5 gene regulation using bacterial artificial chromosomes. AB - The regulatory regions surrounding many genes may be large and difficult to study using standard transgenic approaches. Here we describe the use of bacterial artificial chromosome clones to rapidly survey hundreds of kilobases of DNA for potential regulatory sequences surrounding the mouse bone morphogenetic protein-5 (Bmp5) gene. Simple coinjection of large insert clones with lacZ reporter constructs recapitulates all of the sites of expression observed previously with numerous small constructs covering a large, complex regulatory region. The coinjection approach has made it possible to rapidly survey other regions of the Bmp5 gene for potential control elements, to confirm the location of several elements predicted from previous expression studies using regulatory mutations at the Bmp5 locus, to test whether Bmp5 control regions act similarly on endogenous and foreign promoters, and to show that Bmp5 control elements are capable of rescuing phenotypic effects of a Bmp5 deficiency. This rapid approach has identified new Bmp5 control regions responsible for controlling the development of specific anatomical structures in the vertebrate skeleton. A similar approach may be useful for studying complex control regions surrounding many other genes important in embryonic development and human disease. PMID- 10677508 TI - Zic2 regulates the kinetics of neurulation. AB - Mutation in human ZIC2, a zinc finger protein homologous to Drosophila odd paired, causes holoprosencephaly (HPE), which is a common, severe malformation of the brain in humans. However, the pathogenesis is largely unknown. Here we show that reduced expression (knockdown) of mouse Zic2 causes neurulation delay, resulting in HPE and spina bifida. Differentiation of the most dorsal neural plate, which gives rise to both roof plate and neural crest cells, also was delayed as indicated by the expression lag of a roof plate marker, Wnt3a. In addition the development of neural crest derivatives such as dorsal root ganglion was impaired. These results suggest that the Zic2 expression level is crucial for the timing of neurulation. Because the Zic2 knockdown mouse is the first mutant with HPE and spina bifida to survive to the perinatal period, the mouse will promote analyses of not only the neurulation but also the pathogenesis of human HPE. PMID- 10677509 TI - Endogenous expression of Mullerian inhibiting substance in early postnatal rat sertoli cells requires multiple steroidogenic factor-1 and GATA-4-binding sites. AB - Mullerian inhibiting substance (MIS) is a key element required to complete mammalian male sex differentiation. The expression pattern of MIS is tightly regulated in fetal, neonatal, and prepubertal testes and adult ovaries and is well conserved among mammalian species. Although several factors have been shown to be essential to MIS expression, its regulatory mechanisms are not fully understood. We have examined MIS promoter activity in 2-day postnatal primary cultures of rat Sertoli cells that continue to express endogenous MIS mRNA. Using this system, we found that the region between human MIS-269 and -192 is necessary for full MIS promoter activity. We identified by DNase I footprint and electrophoretic mobility-shift analyses a distal steroidogenic factor-1 (SF-1) binding site that is essential for full promoter activity. Mutational analysis of this new distal SF-1 site and the previously identified proximal SF-1 site showed that both are necessary for transcriptional activation. Moreover, the proximal promoter also contains multiple GATA-4-binding sites that are essential for functional promoter activity. Thus multiple SF-1- and GATA-4-binding sites in the MIS promoter are required for normal tissue-specific and developmental expression of MIS. PMID- 10677510 TI - Climate change is affecting altitudinal migrants and hibernating species. AB - Calendar date of the beginning of the growing season at high altitude in the Colorado Rocky Mountains is variable but has not changed significantly over the past 25 years. This result differs from growing evidence from low altitudes that climate change is resulting in a longer growing season, earlier migrations, and earlier reproduction in a variety of taxa. At our study site, the beginning of the growing season is controlled by melting of the previous winter's snowpack. Despite a trend for warmer spring temperatures the average date of snowmelt has not changed, perhaps because of the trend for increased winter precipitation. This disjunction between phenology at low and high altitudes may create problems for species, such as many birds, that migrate over altitudinal gradients. We present data indicating that this already may be true for American robins, which are arriving 14 days earlier than they did in 1981; the interval between arrival date and the first date of bare ground has grown by 18 days. We also report evidence for an effect of climate change on hibernation behavior; yellow-bellied marmots are emerging 38 days earlier than 23 years ago, apparently in response to warmer spring air temperatures. Migrants and hibernators may experience problems as a consequence of these changes in phenology, which may be exacerbated if climate models are correct in their predictions of increased winter snowfall in our study area. The trends we report for earlier formation of permanent snowpack and for a longer period of snow cover also have implications for hibernating species. PMID- 10677511 TI - Chemical defense against predation in an insect egg. AB - The larva of the green lacewing (Ceraeochrysa cubana) (Neuroptera, Chrysopidae) is a natural predator of eggs of Utetheisa ornatrix (Lepidoptera, Arctiidae), a moth that sequesters pyrrolizidine alkaloids from its larval foodplant (Fabaceae, Crotalaria spp.). Utetheisa eggs are ordinarily endowed with the alkaloid. Alkaloid-free Utetheisa eggs, produced experimentally, are pierced by the larva with its sharp tubular jaws and sucked out. Alkaloid-laden eggs, in contrast, are rejected. When attacking an Utetheisa egg cluster (numbering on average 20 eggs), the larva subjects it to an inspection process. It prods and/or pierces a small number of eggs (on average two to three) and, if these contain alkaloid, it passes "negative judgement" on the remainder of the cluster and turns away. Such generalization on the part of the larva makes sense, because the eggs within clusters differ little in alkaloid content. There is, however, considerable between-cluster variation in egg alkaloid content, so clusters in nature can be expected to range widely in palatability. To check each cluster for acceptability must therefore be adaptive for the larva, just as it must be adaptive for Utetheisa to lay its eggs in large clusters and to apportion alkaloid evenly among eggs of a cluster. PMID- 10677512 TI - Biodiversity of Costa Rican salamanders: implications of high levels of genetic differentiation and phylogeographic structure for species formation. AB - Although salamanders are characteristic amphibians in Holarctic temperate habitats, in tropical regions they have diversified evolutionarily only in tropical America. An adaptive radiation centered in Middle America occurred late in the history of a single clade, the supergenus Bolitoglossa (Plethodontidae), and large numbers of species now occur in diverse habitats. Sublineages within this clade decrease in number from the northern to southern parts of Middle America, and in Costa Rica, there are but three. Despite this phylogenetic constraint, Costa Rica has many species; the number of salamander species on one local elevational transect in the Cordillera de Talamanca may be the largest for any such transect in the world. Extraordinary variation in sequences of the mitochondrial gene cytochrome b within a clade of the genus Bolitoglossa in Costa Rica reveals strong phylogeographic structure within a single species, Bolitoglossa pesrubra. Allozymic variation in 19 proteins reveals a pattern largely concordant with the mitochondrial DNA phylogeography. More species exist than are currently recognized. Diversification occurs in restricted geographic areas and involves sharp geographic and elevational differentiation and zonation. In their degree of genetic differentiation at a local scale, these species of the deep tropics exceed the known variation of extratropical salamanders, which also differ in being less restricted in elevational range. Salamanders display "tropicality" in that although speciose, they are usually local in distribution and rare. They display strong ecological and physiological differentiation that may contribute importantly to morphological divergence and species formation. PMID- 10677513 TI - Population genetics of ice age brown bears. AB - The Pleistocene was a dynamic period for Holarctic mammal species, complicated by episodes of glaciation, local extinctions, and intercontinental migration. The genetic consequences of these events are difficult to resolve from the study of present-day populations. To provide a direct view of population genetics in the late Pleistocene, we measured mitochondrial DNA sequence variation in seven permafrost-preserved brown bear (Ursus arctos) specimens, dated from 14,000 to 42,000 years ago. Approximately 36,000 years ago, the Beringian brown bear population had a higher genetic diversity than any extant North American population, but by 15,000 years ago genetic diversity appears similar to the modern day. The older, genetically diverse, Beringian population contained sequences from three clades now restricted to local regions within North America, indicating that current phylogeographic patterns may provide misleading data for evolutionary studies and conservation management. The late Pleistocene phylogeographic data also indicate possible colonization routes to areas south of the Cordilleran ice sheet. PMID- 10677514 TI - Hox cluster genomics in the horn shark, Heterodontus francisci. AB - Reconstructing the evolutionary history of Hox cluster origins will lead to insights into the developmental and evolutionary significance of Hox gene clusters in vertebrate phylogeny and to their role in the origins of various vertebrate body plans. We have isolated two Hox clusters from the horn shark, Heterodontus francisci. These have been sequenced and compared with one another and with other chordate Hox clusters. The results show that one of the horn shark clusters (HoxM) is orthologous to the mammalian HoxA cluster and shows a structural similarity to the amphioxus cluster, whereas the other shark cluster (HoxN) is orthologous to the mammalian HoxD cluster based on cluster organization and a comparison with noncoding and Hox gene-coding sequences. The persistence of an identifiable HoxA cluster over an 800-million-year divergence time demonstrates that the Hox gene clusters are highly integrated and structured genetic entities. The data presented herein identify many noncoding sequence motifs conserved over 800 million years that may function as genetic control motifs essential to the developmental process. PMID- 10677515 TI - Identification of hepatitis B virus indigenous to chimpanzees. AB - Hepatitis B viruses (HBV) and related viruses, classified in the Hepadnaviridae family, are found in a wide variety of mammals and birds. Although the chimpanzee has been the primary experimental model of HBV infection, this species has not been considered a natural host for the virus. Retrospective analysis of 13 predominantly wild-caught chimpanzees with chronic HBV infection identified a unique chimpanzee HBV strain in 11 animals. Nucleotide and derived amino acid analysis of the complete HBV genome and the gene coding for the hepatitis B surface antigen (S gene) identified sequence patterns that could be used to reliably identify chimpanzee HBV. This analysis indicated that chimpanzee HBV is distinct from known human HBV genotypes and is closely related to HBVs previously isolated from a chimpanzee, gibbons, gorillas, and orangutans. PMID- 10677516 TI - In vitro cloning of complex mixtures of DNA on microbeads: physical separation of differentially expressed cDNAs. AB - We describe a method for cloning nucleic acid molecules onto the surfaces of 5 micrometer microbeads rather than in biological hosts. A unique tag sequence is attached to each molecule, and the tagged library is amplified. Unique tagging of the molecules is achieved by sampling a small fraction (1%) of a very large repertoire of tag sequences. The resulting library is hybridized to microbeads that each carry approximately 10(6) strands complementary to one of the tags. About 10(5) copies of each molecule are collected on each microbead. Because such clones are segregated on microbeads, they can be operated on simultaneously and then assayed separately. To demonstrate the utility of this approach, we show how to label and extract microbeads bearing clones differentially expressed between two libraries by using a fluorescence-activated cell sorter (FACS). Because no prior information about the cloned molecules is required, this process is obviously useful where sequence databases are incomplete or nonexistent. More importantly, the process also permits the isolation of clones that are expressed only in given tissues or that are differentially expressed between normal and diseased states. Such clones then may be spotted on much more cost-effective, tissue- or disease-directed, low-density planar microarrays. PMID- 10677517 TI - Mitochondrial carbonic anhydrase CA VB: differences in tissue distribution and pattern of evolution from those of CA VA suggest distinct physiological roles. AB - A cDNA for a second mouse mitochondrial carbonic anhydrase (CA) called CA VB was identified by homology to the previously characterized murine CA V, now called CA VA. The full-length cDNA encodes a 317-aa precursor that contains a 33-aa classical mitochondrial leader sequence. Comparison of products expressed from cDNAs for murine CA VB and CA VA in COS cells revealed that both expressed active CAs that localized in mitochondria, and showed comparable activities in crude extracts and in mitochondria isolated from transfected COS cells. Northern blot analyses of total RNAs from mouse tissues and Western blot analyses of mouse tissue homogenates showed differences in tissue-specific expression between CA VB and CA VA. CA VB was readily detected in most tissues, while CA VA expression was limited to liver, skeletal muscle, and kidney. The human orthologue of murine CA VB was recently reported also. Comparison of the CA domain sequence of human CA VB with that reported here shows that the CA domains of CA VB are much more highly conserved between mouse and human (95% identity) than the CA domains of mouse and human CA VAs (78% identity). Analysis of phylogenetic relationships between these and other available human and mouse CA isozyme sequences revealed that mammalian CA VB evolved much more slowly than CA VA, accepting amino acid substitutions at least 4.5 times more slowly since each evolved from its respective human-mouse ancestral gene around 90 million years ago. Both the differences in tissue distribution and the much greater evolutionary constraints on CA VB sequences suggest that CA VB and CA VA have evolved to assume different physiological roles. PMID- 10677518 TI - Nonlinearity in genetic decoding: homologous DNA replicase genes use alternatives of transcriptional slippage or translational frameshifting. AB - The tau and gamma subunits of DNA polymerase III are both encoded by a single gene in Escherichia coli and Thermus thermophilus. gamma is two-thirds the size of tau and shares virtually all its amino acid sequence with tau. E. coli and T. thermophilus have evolved very different mechanisms for setting the approximate 1:1 ratio between tau and gamma. Both mechanisms put ribosomes into alternate reading frames so that stop codons in the new frame serve to make the smaller gamma protein. In E. coli, approximately 50% of initiating ribosomes translate the dnaX mRNA conventionally to give tau, but the other 50% shift into the -1 reading frame at a specific site (A AAA AAG) in the mRNA to produce gamma. In T. thermophilus ribosomal frameshifting is not required: the dnaX mRNA is a heterogeneous population of molecules with different numbers of A residues arising from transcriptional slippage on a run of nine T residues in the DNA template. Translation of the subpopulation containing nine As (or +/- multiples of three As) yields tau. The rest of the population of mRNAs (containing nine +/- nonmultiples of three As) puts ribosomes into the alternate reading frames to produce the gamma protein(s). It is surprising that two rather similar dnaX sequences in E. coli and T. thermophilus lead to very different mechanisms of expression. PMID- 10677519 TI - Protective DNA vaccination against organ-specific autoimmunity is highly specific and discriminates between single amino acid substitutions in the peptide autoantigen. AB - DNA vaccines that encode encephalitogenic sequences in tandem can protect from subsequent experimental autoimmune encephalomyelitis induced with the corresponding peptide. The mechanism for this protection and, in particular, if it is specific for the amino acid sequence encoding the vaccine are not known. We show here that a single amino acid exchange in position 79 from serine (nonself) to threonine (self) in myelin basic protein peptide MBP68-85, which is a major encephalitogenic determinant for Lewis rats, dramatically alters the protection. Moreover, vaccines encoding the encephalitogenic sequence MBP68-85 do not protect against the second encephalitogenic sequence MBP89-101 in Lewis rats and vice versa. Thus, protective immunity conferred by DNA vaccination exquisitely discriminates between peptide target autoantigens. No bystander suppression was observed. The exact underlying mechanisms remain elusive because no simple correlation between impact on ex vivo responses and protection against disease were noted. PMID- 10677520 TI - Human cytomegalovirus harbors its own unique IL-10 homolog (cmvIL-10). AB - We identified a viral IL-10 homolog encoded by an ORF (UL111a) within the human cytomegalovirus (CMV) genome, which we designated cmvIL-10. cmvIL-10 can bind to the human IL-10 receptor and can compete with human IL-10 for binding sites, despite the fact that these two proteins are only 27% identical. cmvIL-10 requires both subunits of the IL-10 receptor complex to induce signal transduction events and biological activities. The structure of the cmvIL-10 gene is unique by itself. The gene retained two of four introns of the IL-10 gene, but the length of the introns was reduced. We demonstrated that cmvIL-10 is expressed in CMV-infected cells. Thus, expression of cmvIL-10 extends the range of counter measures developed by CMV to circumvent detection and destruction by the host immune system. PMID- 10677521 TI - Early expression of antiinsulin autoantibodies of humans and the NOD mouse: evidence for early determination of subsequent diabetes. AB - With the development of an insulin autoantibody (IAA) assay performed in 96-well filtration plates, we have evaluated prospectively the development of IAA in NOD mice (from 4 weeks of age) and children (from 7 to 10 months of age) at genetic risk for the development of type 1 diabetes. NOD mice had heterogeneous expression of IAA despite being inbred. IAA reached a peak between 8 and 16 weeks and then declined. IAA expression by NOD mice at 8 weeks of age was strongly associated with early development of diabetes, which occurred at 16-18 weeks of age (NOD mice IAA(+) at 8 weeks: 83% (5/6) diabetic by 18 weeks versus 11% (1/9) of IAA negative at 8 weeks; P <.01). In man, IAA was frequently present as early as 9 months of age, the first sampling time. Of five children found to have persistent IAA before 1 year of age, four have progressed to diabetes (all before 3.5 years of age) and the fifth is currently less than age 2. Of the 929 children not expressing persistent IAA before age 1, only one has progressed to diabetes to date (age onset 3), and this child expressed IAA at his second visit (age 1.1). In new onset patients, the highest levels of IAA correlated with an earlier age of diabetes onset. Our data suggest that the program for developing diabetes of NOD mice and humans is relatively "fixed" early in life and, for NOD mice, a high risk of early development of diabetes is often determined by 8 weeks of age. With such early determination of high risk of progression to diabetes, immunologic therapies in humans may need to be tested in children before the development of IAA for maximal efficacy. PMID- 10677522 TI - The dual functions of fas ligand in the regulation of peripheral CD8+ and CD4+ T cells. AB - Although Fas ligand (FasL) is well characterized for its capacity to deliver a death signal through its receptor Fas, recent work demonstrates that FasL also can receive signals facilitating antigen (Ag)-specific proliferation of CD8(+) T cells. The fact that the gld mutation differentially influences the proliferative capacity of CD8(+) and CD4(+) T cells presented the intriguing possibility that a single molecule may play opposing roles in these two subpopulations. The present study focuses on how these positive and negative regulatory roles are balanced. We show that naive CD4(+) T cells are responsive to FasL-mediated costimulation on encounter with Ag when Fas-mediated death is prevented. Thus, the machinery responsible for transducing the FasL positive reverse signal operates in both CD4(+) and CD8(+) T cells. Instead, differential control of FasL expression distinguishes the role of FasL in these two T cell subpopulations. FasL costimulation occurs immediately on T cell receptor ligation and correlates with the up-regulation of FasL expression on CD8(+) and naive CD4(+) T cells, both of which are sensitive to the FasL costimulatory signal. Conversely, FasL-initiated death occurs late in an immune response when high levels of FasL expression are maintained on CD4(+) T cells that are sensitive to Fas-mediated death, but not on CD8(+) T cells that are relatively insensitive to this signal. This careful orchestration of FasL expression during times of susceptibility to costimulation and conversely, to death, endows FasL with the capacity to both positively and negatively regulate the peripheral T cell compartment. PMID- 10677523 TI - Role of Syk in B-cell development and antigen-receptor signaling. AB - Antigen receptors (BCRs) on developing B lymphocytes play two opposing roles promoting survival of cells that may later bind a foreign antigen and inhibiting survival of cells that bind too strongly to self-antigens. It is not known how these opposing outcomes are signaled by BCRs on immature B cells. Here we analyze the effect of a null mutation in the Syk tyrosine kinase on maturing B cells displaying a transgene-encoded BCR that binds hen egg lysozyme (HEL). In the absence of HEL antigen, HEL-specific BCRs are expressed normally on the surface of Syk-deficient immature B-lineage cells, but this fails to promote maturation beyond the earliest stages of B-lineage commitment. Binding of HEL antigen, nevertheless, triggers phosphorylation of CD79alpha/beta BCR subunits and modulation of receptors from the surface in Syk-deficient cells, but it cannot induce an intracellular calcium response. Continuous binding of low- or high avidity forms of HEL, expressed as self-antigens, fails to restore the signal needed for maturation. Compared with the effects in the same system of null mutations in other BCR signaling elements, such as CD45 and Lyn kinase, these results indicate that Syk is essential for transmitting a signal that initiates the program of B-lymphocyte maturation. PMID- 10677524 TI - Hypothyroidism in transgenic mice expressing IFN-gamma in the thyroid. AB - IFN-gamma has been implicated with contradictory results in the pathogenetic process of autoimmune (Hashimoto's) thyroiditis, the most common cause of hypothyroidism in adults. To test whether the local production of IFN-gamma can lead to thyroid dysfunction, we have generated transgenic mice that express constitutively IFN-gamma in the thyroid follicular cells. This expression resulted in severe hypothyroidism, with growth retardation and disruption of the thyroid architecture. The hypothyroidism derived from a profound inhibition of the expression of the sodium iodide symporter gene. Taken together, these results indicate a direct role of IFN-gamma in the thyroid dysfunction that occurs in autoimmune thyroiditis. PMID- 10677525 TI - Adapter proteins SLP-76 and BLNK both are expressed by murine macrophages and are linked to signaling via Fcgamma receptors I and II/III. AB - The SLP-76 (Src homology 2 domain-containing leukocyte protein of 76 kDa) adapter protein is expressed in T cells and myeloid cells, whereas its homologue BLNK (B cell linker protein) is expressed in B cells. SLP-76 and BLNK link immunoreceptor tyrosine-based activation motif-containing receptors to signaling molecules that include phospholipase C-gamma, mitogen-activated protein kinases, and the GTPases Ras and Rho. SLP-76 plays a critical role in T cell receptor, FcvarepsilonRI and gpVI collagen receptor signaling, and participates in signaling via FcgammaR and killer cell inhibitory receptors. BLNK plays a critical role in B cell receptor signaling. We show that murine bone marrow-derived macrophages express both SLP 76 and BLNK. Selective ligation of FcgammaRI and FcgammaRII/III resulted in tyrosine phosphorylation of both SLP-76 and BLNK. SLP-76(-/-) bone marrow-derived macrophages display FcgammaR-mediated tyrosine phosphorylation of Syk, phospholipase C-gamma2, and extracellular signal regulated kinases 1 and 2, and normal FcgammaR-dependent phagocytosis. These data suggest that both SLP-76 and BLNK are coupled to FcgammaR signaling in murine macrophages. PMID- 10677526 TI - Polycystin 1 is required for the structural integrity of blood vessels. AB - Autosomal dominant polycystic kidney disease (ADPKD), often caused by mutations in the PKD1 gene, is associated with life-threatening vascular abnormalities that are commonly attributed to the frequent occurrence of hypertension. A previously reported targeted mutation of the mouse homologue of PKD1 was not associated with vascular fragility, leading to the suggestion that the vascular lesion may be of a secondary nature. Here we demonstrate a primary role of PKD1 mutations in vascular fragility. Mouse embryos homozygous for the mutant allele (Pkd1(L)) exhibit s.c. edema, vascular leaks, and rupture of blood vessels, culminating in embryonic lethality at embryonic day 15.5. Kidney and pancreatic ductal cysts are present. The Pkd1-encoded protein, mouse polycystin 1, was detected in normal endothelium and the surrounding vascular smooth muscle cells. These data reveal a requisite role for polycystin 1 in maintaining the structural integrity of the vasculature as well as epithelium and suggest that the nature of the PKD1 mutation contributes to the phenotypic variance in ADPKD. PMID- 10677527 TI - Regulated expression of P210 Bcr-Abl during embryonic stem cell differentiation stimulates multipotential progenitor expansion and myeloid cell fate. AB - P210 Bcr-Abl is an activated tyrosine kinase oncogene encoded by the Philadelphia chromosome associated with human chronic myelogenous leukemia (CML). The disease represents a clonal disorder arising in the pluripotent hematopoietic stem cell. During the chronic phase, patients present with a dramatic expansion of myeloid cells and a mild anemia. Retroviral gene transfer and transgenic expression in rodents have demonstrated the ability of Bcr-Abl to induce various types of leukemia. However, study of human CML or rodent models has not determined the direct and immediate effects of Bcr-Abl on hematopoietic cells from those requiring secondary genetic or epigenetic changes selected during the pathogenic process. We utilized tetracycline-regulated expression of Bcr-Abl from a promoter engineered for robust expression in primitive stem cells through multilineage blood cell development in combination with the in vitro differentiation of embryonal stem cells into hematopoietic elements. Our results demonstrate that Bcr-Abl expression alone is sufficient to increase the number of multipotent and myeloid lineage committed progenitors in a dose-dependent manner while suppressing the development of committed erythroid progenitors. These effects are reversible upon extinguishing Bcr-Abl expression. These findings are consistent with Bcr-Abl being the sole genetic change needed for the establishment of the chronic phase of CML and provide a powerful system for the analysis of any genetic change that alters cell growth and lineage choices of the hematopoietic stem cell. PMID- 10677528 TI - Cardiolipin is a normal component of human plasma lipoproteins. AB - Anticardiolipin (anti-CL) antibodies, diagnostic for antiphospholipid antibody syndrome, are associated with increased risks of venous and arterial thrombosis. Because CL selectively enhances activated protein C/protein S-dependent anticoagulant activities in purified systems and because CL is not known to be a normal plasma component, we searched for CL in plasma. Plasma lipid extracts [chloroform/methanol (2:1, vol/vol)] were subjected to analyses by using TLC, analytical HPLC, and MS. A plasma lipid component was purified that was indistinguishable from reference CL (M:1448). When CL in 40 fasting plasma lipid extracts (20 males, 20 females) was quantitated by using HPLC, CL (mean +/- SD) was 14.9 +/- 3.7 microgram/ml (range 9.1 to 24.2) and CL was not correlated with phosphatidylserine (3.8 +/- 1.7 microgram/ml), phosphatidylethanolamine (64 +/- 20 microgram/ml), or choline-containing phospholipid (1,580 +/- 280 microgram/ml). Based on studies of fasting blood donors, CL (>/=94%) was recovered in very low density, low density, and high density lipoproteins (11 +/- 5.3%, 67 +/- 11.0%, and 17 +/- 10%, respectively), showing that the majority of plasma CL (67%) is in low density lipoprotein. Analysis of relative phospholipid contents of lipoproteins indicated that high density lipoprotein is selectively enriched in CL and phosphatidylethanolamine. These results shows that CL is a normal plasma component and suggest that the epitopes of antiphospholipid antibodies could include CL or oxidized CL in lipoproteins or in complexes with plasma proteins (e. g., beta(2)-glycoprotein I, prothrombin, protein C, or protein S) or with platelet or endothelial surface proteins. PMID- 10677529 TI - Phosphatidylinositol 3-kinase signaling mediates angiogenesis and expression of vascular endothelial growth factor in endothelial cells. AB - Phosphatidylinositol 3-kinase (PI 3-kinase) is a signaling molecule that controls numerous cellular properties and activities. The oncogene v-p3k is a homolog of the gene coding for the catalytic subunit of PI 3-kinase, p110alpha. P3k induces transformation of cells in culture, formation of hemangiosarcomas in young chickens, and myogenic differentiation in myoblasts. Here, we describe a role of PI 3-kinase in angiogenesis. Overexpression of the v-P3k protein or of cellular PI 3-kinase equipped with a myristylation signal, Myr-P3k, can induce angiogenesis in the chorioallantoic membrane (CAM) of the chicken embryo. This process is characterized by extensive sprouting of new blood vessels and enlargement of preexisting vessels. Overexpression of the myristylated form of the PI 3-kinase target Akt, Myr-Akt, also induces angiogenesis. Overexpression of the tumor suppressor PTEN or of dominant-negative constructs of PI 3-kinase inhibits angiogenesis in the yolk sac of chicken embryos, suggesting that PI 3 kinase and Akt signaling is required for normal embryonal angiogenesis. The levels of mRNA for vascular endothelial growth factor (VEGF) are elevated in cells expressing activated PI 3-kinase or Myr-Akt. VEGF mRNA levels are also increased by insulin treatment through the PI 3-kinase-dependent pathway. VEGF mRNA levels are decreased in cells treated with the PI 3-kinase inhibitor LY294002 and restored by overexpression of v-P3k or Myr-Akt. Overexpression of VEGF by the RCAS vector induces angiogenesis in chicken embryos. These results suggest that PI 3-kinase plays an important role in angiogenesis and regulates VEGF expression. PMID- 10677530 TI - Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent endogenous activator of the cell death pathway and functions by activating the cell surface death receptors 4 and 5 (DR4 and DR5). TRAIL is nontoxic in vivo and preferentially kills neoplastically transformed cells over normal cells by an undefined mechanism. Radiotherapy is a common treatment for breast cancer as well as many other cancers. Here we demonstrate that ionizing radiation can sensitize breast carcinoma cells to TRAIL-induced apoptosis. This synergistic effect is p53 dependent and may be the result of radiation-induced up-regulation of the TRAIL receptor DR5. Importantly, TRAIL and ionizing radiation have a synergistic effect in the regression of established breast cancer xenografts. Changes in tumor cellularity and extracellular space were monitored in vivo by diffusion-weighted magnetic resonance imaging (diffusion MRI), a noninvasive technique to produce quantitative images of the apparent mobility of water within a tissue. Increased water mobility was observed in combined TRAIL- and radiation-treated tumors but not in tumors treated with TRAIL or radiation alone. Histological analysis confirmed the loss of cellularity and increased numbers of apoptotic cells in TRAIL- and radiation-treated tumors. Taken together, our results provide support for combining radiation with TRAIL to improve tumor eradication and suggest that efficacy of apoptosis-inducing cancer therapies may be monitored noninvasively, using diffusion MRI. PMID- 10677531 TI - Human AML1/MDS1/EVI1 fusion protein induces an acute myelogenous leukemia (AML) in mice: a model for human AML. AB - The human t(3;21)(q26;q22) translocation is found as a secondary mutation in some cases of chronic myelogenous leukemia during the blast phase and in therapy related myelodysplasia and acute myelogenous leukemia. One result of this translocation is a fusion between the AML1, MDS1, and EVI1 genes, which encodes a transcription factor of approximately 200 kDa. The role of the AML1/MDS1/EVI1 (AME) fusion gene in leukemogenesis is largely unknown. In this study, we analyzed the effect of the AME fusion gene in vivo by expressing it in mouse bone marrow cells via retroviral transduction. We found that mice transplanted with AME-transduced bone marrow cells suffered from an acute myelogenous leukemia (AML) 5-13 mo after transplantation. The disease could be readily transferred into secondary recipients with a much shorter latency. Morphological analysis of peripheral blood and bone marrow smears demonstrated the presence of myeloid blast cells and differentiated but immature cells of both myelocytic and monocytic lineages. Cytochemical and flow cytometric analysis confirmed that these mice had a disease similar to the human acute myelomonocytic leukemia. This murine model for AME-induced AML will help dissect the molecular mechanism of AML and the molecular biology of the AML1, MDS1, and EVI1 genes. PMID- 10677532 TI - Identification of a Plasmodium falciparum intercellular adhesion molecule-1 binding domain: a parasite adhesion trait implicated in cerebral malaria. AB - Binding of infected erythrocytes to brain venules is a central pathogenic event in the lethal malaria disease complication, cerebral malaria. The only parasite adhesion trait linked to cerebral sequestration is binding to intercellular adhesion molecule-1 (ICAM-1). In this report, we show that Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) binds ICAM-1. We have cloned and expressed PfEMP1 recombinant proteins from the A4tres parasite. Using heterologous expression in mammalian cells, the minimal ICAM-1 binding domain was a complex domain consisting of the second Duffy binding-like (DBL) domain and the C2 domain. Constructs that contained either domain alone did not bind ICAM-1. Based on phylogenetic criteria, there are five distinct PfEMP1 DBL types designated alpha, beta, gamma, delta, and epsilon. The DBL domain from the A4tres that binds ICAM-1 is DBLbeta type. A PfEMP1 cloned from a distinct ICAM-1 binding variant, the A4 parasite, contains a DBLbeta domain and a C2 domain in tandem arrangement similar to the A4tres PfEMP1. Anti-PfEMP1 antisera implicate the DBLbeta domain from A4var PfEMP1 in ICAM-1 adhesion. The identification of a P. falciparum ICAM-1 binding domain may clarify mechanisms responsible for the pathogenesis of cerebral malaria and lead to interventions or vaccines that reduce malarial disease. PMID- 10677533 TI - Generation of in vivo activating factors in the ischemic intestine by pancreatic enzymes. AB - One of the early events in physiological shock is the generation of activators for leukocytes, endothelial cells, and other cells in the cardiovascular system. The mechanism by which these activators are produced has remained unresolved. We examine here the hypothesis that pancreatic digestive enzymes in the ischemic intestine may be involved in the generation of activators during intestinal ischemia. The lumen of the small intestine of rats was continuously perfused with saline containing a broadly acting pancreatic enzyme inhibitor (6-amidino-2 naphthyl p-guanidinobenzoate dimethanesulfate, 0.37 mM) before and during ischemia of the small intestine by splanchnic artery occlusion. This procedure inhibited activation of circulating leukocytes during occlusion and reperfusion. It also prevented the appearance of activators in portal venous and systemic artery plasma and attenuated initiating symptoms of multiple organ injury in shock. Intestinal tissue produces only low levels of activators in the absence of pancreatic enzymes, whereas in the presence of enzymes, activators are produced in a concentration- and time-dependent fashion. The results indicate that pancreatic digestive enzymes in the ischemic intestine serve as an important source for cell activation and inflammation, as well as multiple organ failure. PMID- 10677534 TI - Global analysis of gene expression in pulmonary fibrosis reveals distinct programs regulating lung inflammation and fibrosis. AB - The molecular mechanisms of pulmonary fibrosis are poorly understood. We have used oligonucleotide arrays to analyze the gene expression programs that underlie pulmonary fibrosis in response to bleomycin, a drug that causes lung inflammation and fibrosis, in two strains of susceptible mice (129 and C57BL/6). We then compared the gene expression patterns in these mice with 129 mice carrying a null mutation in the epithelial-restricted integrin beta6 subunit (beta6(-/-)), which develop inflammation but are protected from pulmonary fibrosis. Cluster analysis identified two distinct groups of genes involved in the inflammatory and fibrotic responses. Analysis of gene expression at multiple time points after bleomycin administration revealed sequential induction of subsets of genes that characterize each response. The availability of this comprehensive data set should accelerate the development of more effective strategies for intervention at the various stages in the development of fibrotic diseases of the lungs and other organs. PMID- 10677535 TI - Lipotoxic heart disease in obese rats: implications for human obesity. AB - To determine the mechanism of the cardiac dilatation and reduced contractility of obese Zucker Diabetic Fatty rats, myocardial triacylglycerol (TG) was assayed chemically and morphologically. TG was high because of underexpression of fatty acid oxidative enzymes and their transcription factor, peroxisome proliferator activated receptor-alpha. Levels of ceramide, a mediator of apoptosis, were 2-3 times those of controls and inducible nitric oxide synthase levels were 4 times greater than normal. Myocardial DNA laddering, an index of apoptosis, reached 20 times the normal level. Troglitazone therapy lowered myocardial TG and ceramide and completely prevented DNA laddering and loss of cardiac function. In this paper, we conclude that cardiac dysfunction in obesity is caused by lipoapoptosis and is prevented by reducing cardiac lipids. PMID- 10677536 TI - Chemical chaperones mediate increased secretion of mutant alpha 1-antitrypsin (alpha 1-AT) Z: A potential pharmacological strategy for prevention of liver injury and emphysema in alpha 1-AT deficiency. AB - In alpha1-AT deficiency, a misfolded but functionally active mutant alpha1-ATZ (alpha1-ATZ) molecule is retained in the endoplasmic reticulum of liver cells rather than secreted into the blood and body fluids. Emphysema is thought to be caused by the lack of circulating alpha1-AT to inhibit neutrophil elastase in the lung. Liver injury is thought to be caused by the hepatotoxic effects of the retained alpha1-ATZ. In this study, we show that several "chemical chaperones," which have been shown to reverse the cellular mislocalization or misfolding of other mutant plasma membrane, nuclear, and cytoplasmic proteins, mediate increased secretion of alpha1-ATZ. In particular, 4-phenylbutyric acid (PBA) mediated a marked increase in secretion of functionally active alpha1-ATZ in a model cell culture system. Moreover, oral administration of PBA was well tolerated by PiZ mice (transgenic for the human alpha1-ATZ gene) and consistently mediated an increase in blood levels of human alpha1-AT reaching 20-50% of the levels present in PiM mice and normal humans. Because clinical studies have suggested that only partial correction is needed for prevention of both liver and lung injury in alpha1-AT deficiency and PBA has been used safely in humans, it constitutes an excellent candidate for chemoprophylaxis of target organ injury in alpha1-AT deficiency. PMID- 10677537 TI - Cure of human carcinoma xenografts by a single dose of pretargeted yttrium-90 with negligible toxicity. AB - A covalent conjugate (NR-LU-10/SA) was prepared between streptavidin (SA) and NR LU-10, a mAb that binds an antigen expressed on the surface of most human carcinomas. NR-LU-10/SA was injected into nude mice bearing human tumor xenografts. Injection of biotinylated galactosyl-human serum albumin reduced the circulating levels of conjugate by 95%. Subsequent administration of (90)Y-1,4,7, 10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin achieved peak uptake at the tumor within 2 hr while >80% of the radioactivity was eliminated in the urine. A single dose of 600-800 microCi of (90)Y-1,4,7,10-tetraazacyclododecane 1,4,7,10-tetraacetic acid-biotin produced cures in 10/10 mice with established (>200 mm(3)) s.c. human small cell lung or colon cancer xenografts and 8/10 cures in mice with human breast cancer xenografts without significant toxicity. PMID- 10677538 TI - Facilitated diffusion of fructose via the phosphoenolpyruvate/glucose phosphotransferase system of Escherichia coli. AB - From mutants of Escherichia coli unable to utilize fructose via the phosphoenolpyruvate/glycose phosphotransferase system (PTS), further mutants were selected that grow on fructose as the sole carbon source, albeit with relatively low affinity for that hexose (K(m) for growth approximately 8 mM but with V(max) for generation time approximately 1 h 10 min); the fructose thus taken into the cells is phosphorylated to fructose 6-phosphate by ATP and a cytosolic fructo(manno)kinase (Mak). The gene effecting the translocation of fructose was identified by Hfr-mediated conjugations and by phage-mediated transduction as specifying an isoform of the membrane-spanning enzyme II(Glc) of the PTS, which we designate ptsG-F. Exconjugants that had acquired ptsG(+) from Hfr strains used for mapping (designated ptsG-I) grew very poorly on fructose (V(max) approximately 7 h 20 min), even though they were rich in Mak activity. A mutant of E. coli also rich in Mak but unable to grow on glucose by virtue of transposon mediated inactivations both of ptsG and of the genes specifying enzyme II(Man) (manXYZ) was restored to growth on glucose by plasmids containing either ptsG-F or ptsG-I, but only the former restored growth on fructose. Sequence analysis showed that the difference between these two forms of ptsG, which was reflected also by differences in the rates at which they translocated mannose and glucose analogs such as methyl alpha-glucoside and 2-deoxyglucose, resided in a substitution of G in ptsG-I by T in ptsG-F in the first position of codon 12, with consequent replacement of valine by phenylalanine in the deduced amino acid sequence. PMID- 10677539 TI - A herpes simplex virus 1 recombinant lacking the glycoprotein G coding sequences is defective in entry through apical surfaces of polarized epithelial cells in culture and in vivo. AB - During infection of a new host, the first surfaces encountered by herpes simplex viruses are the apical membranes of epithelial cells of mucosal surfaces. These cells are highly polarized, and the protein composition of their apical and basolateral membranes are very different, so that different viral entry pathways have evolved for each surface. To determine whether the viral glycoprotein G (gG) is specifically required for efficient infection of a particular surface of polarized cells, apical and basal surfaces were infected with wild-type virus or a gG deletion mutant. After infection of polarized cells in culture, the gG(-) virus was deficient in infection of apical surfaces but was able to infect cells through basal membranes, replicate, and spread into surrounding cells. The gG dependent step in apical infection was a stage beyond attachment. After in vivo infection of apical surfaces of epithelial cells of nonscarified mouse corneas, infection by glycoprotein C(-) or gG(-) virus was considerably reduced as compared with that observed after infection with wild-type virus. In contrast, when corneas were scarified, allowing virus access to other cell surfaces, the gG and glycoprotein C deletion mutants infected eyes as efficiently as wild-type viruses. A secondary mutation allowing infection of apical surfaces by gG(-) virus arose readily during passage of the virus in nonpolarized cells, indicating that either the gG-dependent step of apical infection can be bypassed or that another viral protein can acquire the same function. PMID- 10677540 TI - Importance of newly generated neurons in the adult olfactory bulb for odor discrimination. AB - In adult rodents, neurons are continually generated in the subventricular zone of the forebrain, from where they migrate tangentially toward the olfactory bulb, the only known target for these neuronal precursors. Within the main olfactory bulb, they ascend radially into the granule and periglomerular cell layers, where they differentiate mainly into local interneurons. The functional consequences of this permanent generation and integration of new neurons into existing circuits are unknown. To address this question, we used neural cell adhesion molecule deficient mice that have documented deficits in the migration of olfactory-bulb neuron precursors, leading to about 40% size reduction of this structure. Our anatomical study reveals that this reduction is restricted to the granule cell layer, a structure that contains exclusively gamma-aminobutyric acid (GABA)ergic interneurons. Furthermore, mutant mice were subjected to experiments designed to examine the behavioral consequences of such anatomical alteration. We found that the specific reduction in the newly generated interneuron population resulted in an impairment of discrimination between odors. In contrast, both the detection thresholds for odors and short-term olfactory memory were unaltered, demonstrating that a critical number of bulbar granule cells is crucial only for odor discrimination but not for general olfactory functions. PMID- 10677541 TI - Activation of a heterologously expressed octopamine receptor coupled only to adenylyl cyclase produces all the features of presynaptic facilitation in aplysia sensory neurons. AB - Short-term behavioral sensitization of the gill-withdrawal reflex after tail stimuli in Aplysia leads to an enhancement of the connections between sensory and motor neurons of this reflex. Both behavioral sensitization and enhancement of the connection between sensory and motor neurons are importantly mediated by serotonin. Serotonin activates two types of receptors in the sensory neurons, one of which is coupled to the cAMP/protein kinase A (PKA) pathway and the other to the inositol triphosphate/protein kinase C (PKC) pathway. Here we describe a genetic approach to assessing the isolated contribution of the PKA pathway to short-term facilitation. We have cloned from Aplysia an octopamine receptor gene, Ap oa(1), that couples selectively to the cAMP/PKA pathway. We have ectopically expressed this receptor in Aplysia sensory neurons of the pleural ganglia, where it is not normally expressed. Activation of this receptor by octopamine stimulates all four presynaptic events involved in short-term synaptic facilitation that are normally produced by serotonin: (i) membrane depolarization; (ii) increased membrane excitability; (iii) increased spike duration; and (iv) presynaptic facilitation. These results indicate that the cAMP/PKA pathway alone is sufficient to produce all the features of presynaptic facilitation. PMID- 10677542 TI - An electric lobe suppressor for a yeast choline transport mutation belongs to a new family of transporter-like proteins. AB - Choline is an important metabolite in all cells due to the major contribution of phosphatidylcholine to the production of membranes, but it takes on an added role in cholinergic neurons where it participates in the synthesis of the neurotransmitter acetylcholine. We have cloned a suppressor for a yeast choline transport mutation from a Torpedo electric lobe yeast expression library by functional complementation. The full-length clone encodes a protein with 10 putative transmembrane domains, two of which contain transporter-like motifs, and whose expression increased high-affinity choline uptake in mutant yeast. The gene was called CTL1 for its choline transporter-like properties. The homologous rat gene, rCTL1, was isolated and found to be highly expressed as a 3. 5-kb transcript in the spinal cord and brain and as a 5-kb transcript in the colon. In situ hybridization showed strong expression of rCTL1 in motor neurons and oligodendrocytes and to a lesser extent in various neuronal populations throughout the rat brain. High levels of rCTL1 were also identified in the mucosal cell layer of the colon. Although the sequence of the CTL1 gene shows clear homology with a single gene in Caenorhabditis elegans, several homologous genes are found in mammals (CTL2-4). These results establish a new family of genes for transporter-like proteins in eukaryotes and suggest that one of its members, CTL1, is involved in supplying choline to certain cell types, including a specific subset of cholinergic neurons. PMID- 10677543 TI - Columnar distribution of serotonin-dependent plasticity within kitten striate cortex. AB - Recent studies have identified the potential for an important role for serotonin (5-HT) receptors in the developmental plasticity of the kitten visual cortex. 5 HT(2C) receptors are transiently expressed in a patchy fashion in the visual cortex of kittens between 30-80 days of age complementary to patches demarcated by cytochrome oxidase staining. 5-HT, operating via 5-HT(2C) receptors, increases cortical synaptic plasticity as assessed both in brain slices and in vivo. Herein, we report that bath application of 5-HT substantially increases the probability of long-term potentiation within 5-HT(2C) receptor-rich zones of cortex, but this effect is not observed in the 5-HT(2C) receptor-poor zones. Instead, in these zones, 5-HT application increases the probability of long-term depression. These location-specific effects of 5-HT may promote the formation of compartment-specific cortical responses. PMID- 10677544 TI - Poly(ADP-ribosyl)ation basally activated by DNA strand breaks reflects glutamate nitric oxide neurotransmission. AB - Poly(ADP-ribose) polymerase (PARP) transfers ADP ribose groups from NAD(+) to nuclear proteins after activation by DNA strand breaks. PARP overactivation by massive DNA damage causes cell death via NAD(+) and ATP depletion. Heretofore, PARP has been thought to be inactive under basal physiologic conditions. We now report high basal levels of PARP activity and DNA strand breaks in discrete neuronal populations of the brain, in ventricular ependymal and subependymal cells and in peripheral tissues. In some peripheral tissues, such as skeletal muscle, spleen, heart, and kidney, PARP activity is reduced only partially in mice with PARP-1 gene deletion (PARP-1(-/-)), implicating activity of alternative forms of PARP. Glutamate neurotransmission involving N-methyl-d-aspartate (NMDA) receptors and neuronal nitric oxide synthase (nNOS) activity in part mediates neuronal DNA strand breaks and PARP activity, which are diminished by NMDA antagonists and NOS inhibitors and also diminished in mice with targeted deletion of nNOS gene (nNOS(-/-)). An increase in NAD(+) levels after treatment with NMDA antagonists or NOS inhibitors, as well as in nNOS(-/-) mice, indicates that basal glutamate-PARP activity regulates neuronal energy dynamics. PMID- 10677545 TI - Carbon monoxide and nitric oxide as coneurotransmitters in the enteric nervous system: evidence from genomic deletion of biosynthetic enzymes. AB - Nitric oxide (NO) and carbon monoxide (CO) seem to be neurotransmitters in the brain. The colocalization of their respective biosynthetic enzymes, neuronal NO synthase (nNOS) and heme oxygenase-2 (HO2), in enteric neurons and altered intestinal function in mice with genomic deletion of the enzymes (nNOS(Delta/Delta) and HO2(Delta/Delta)) suggest neurotransmitter roles for NO and CO in the enteric nervous system. We now establish that NO and CO are both neurotransmitters that interact as cotransmitters. Small intestinal smooth muscle cells from nNOS(Delta/Delta) and HO2(Delta/Delta) mice are depolarized, with apparent additive effects in the double knockouts (HO2(Delta/Delta)/nNOS(Delta/Delta)). Muscle relaxation and inhibitory neurotransmission are reduced in the mutant mice. In HO2(Delta/Delta) preparations, responses to electrical field stimulation are nearly abolished despite persistent nNOS expression, whereas exogenous CO restores normal responses, indicating that the NO system does not function in the absence of CO generation. PMID- 10677546 TI - Regulation of the phosphorylation of the dopamine- and cAMP-regulated phosphoprotein of 32 kDa in vivo by dopamine D1, dopamine D2, and adenosine A2A receptors. AB - Dopamine D(1), dopamine D(2), and adenosine A(2A) receptors are highly expressed in striatal medium-sized spiny neurons. We have examined, in vivo, the influence of these receptors on the state of phosphorylation of the dopamine- and cAMP regulated phosphoprotein of 32 kDa (DARPP-32). DARPP-32 is a potent endogenous inhibitor of protein phosphatase-1, which plays an obligatory role in dopaminergic transmission. A dose-dependent increase in the state of phosphorylation of DARPP-32 occurred in mouse striatum after systemic administration of the D(2) receptor antagonist eticlopride (0.1-2.0 mg/kg). This effect was abolished in mice in which the gene coding for the adenosine A(2A) receptor was disrupted by homologous recombination. A reduction was also observed in mice that had been pretreated with the selective A(2A) receptor antagonist SCH 58261 (10 mg/kg). The eticlopride-induced increase in DARPP-32 phosphorylation was also decreased by pretreatment with the D(1) receptor antagonist SCH 23390 (0.125 and 0.25 mg/kg) and completely reversed by combined pretreatment with SCH 23390 (0.25 mg/kg) plus SCH 58261 (10 mg/kg). SCH 23390, but not SCH 58261, abolished the increase in DARPP-32 caused by cocaine (15 mg/kg). The results indicate that, in vivo, the state of phosphorylation of DARPP-32 and, by implication, the activity of protein phosphatase-1 are regulated by tonic activation of D(1), D(2), and A(2A) receptors. The results also underscore the fact that the adenosine system plays a role in the generation of responses to dopamine D(2) antagonists in vivo. PMID- 10677547 TI - Role of the cAMP signaling pathway in the regulation of gonadotropin-releasing hormone secretion in GT1 cells. AB - We studied the signaling pathways coupling gonadotropin-releasing hormone (GnRH) secretion to elevations in cAMP levels in the GT1 GnRH-secreting neuronal cell line. We hypothesized that increased cAMP could be acting directly by means of cyclic nucleotide-gated (CNG) cation channels or indirectly by means of activation of cAMP-dependent protein kinase (PKA). We showed that GT1 cells express the three CNG subunits present in olfactory neurons (CNG2, -4.3, and -5) and exhibit functional cAMP-gated cation channels. Activation of PKA does not appear to be necessary for the stimulation of GnRH release by increased levels of cAMP. In fact, pharmacological inhibition of PKA activity caused an increase in the basal secretion of GnRH. Consistent with this observation activation PKA inhibited adenylyl cyclase activity, presumably by inhibiting adenylyl cyclase V expressed in the cells. Therefore, the stimulation of GnRH release by elevations in cAMP appears to be the result of depolarization of the neurons initiated by increased cation conductance by cAMP-gated cation channels. Activation of PKA may constitute a negative-feedback mechanisms for lowering cAMP levels. We hypothesize that these mechanisms could result in oscillations in cAMP levels, providing a biochemical basis for timing the pulsatile release of GnRH. PMID- 10677548 TI - Gamma rhythms and beta rhythms have different synchronization properties. AB - Experimental and modeling efforts suggest that rhythms in the CA1 region of the hippocampus that are in the beta range (12-29 Hz) have a different dynamical structure than that of gamma (30-70 Hz). We use a simplified model to show that the different rhythms employ different dynamical mechanisms to synchronize, based on different ionic currents. The beta frequency is able to synchronize over long conduction delays (corresponding to signals traveling a significant distance in the brain) that apparently cannot be tolerated by gamma rhythms. The synchronization properties are consistent with data suggesting that gamma rhythms are used for relatively local computations whereas beta rhythms are used for higher level interactions involving more distant structures. PMID- 10677550 TI - Dopamine tone regulates D1 receptor trafficking and delivery in striatal neurons in dopamine transporter-deficient mice. AB - In vivo, G protein-coupled receptors (GPCR) for neurotransmitters undergo complex intracellular trafficking that contribute to regulate their abundance at the cell surface. Here, we report a previously undescribed alteration in the subcellular localization of D1 dopamine receptor (D1R) that occurs in vivo in striatal dopaminoceptive neurons in response to chronic and constitutive hyperdopaminergia. Indeed, in mice lacking the dopamine transporter, D1R is in abnormally low abundance at the plasma membrane of cell bodies and dendrites and is largely accumulated in rough endoplasmic reticulum and Golgi apparatus. Decrease of striatal extracellular dopamine concentration with 6-hydroxydopamine (6- OHDA) in heterozygous mice restores delivery of the receptor from the cytoplasm to the plasma membrane in cell bodies. These results demonstrate that, in vivo, in the central nervous system, the storage in cytoplasmic compartments involved in synthesis and the membrane delivery contribute to regulate GPCR availability and abundance at the surface of the neurons under control of the neurotransmitter tone. Such regulation may contribute to modulate receptivity of neurons to their endogenous ligands and related exogenous drugs. PMID- 10677549 TI - Circadian modulation of dopamine receptor responsiveness in Drosophila melanogaster. AB - We investigated the circadian function of Drosophila dopamine receptors by using a behaviorally active decapitated preparation that allows for direct application of drugs to the nerve cord. Quinpirole, a D2-like dopamine receptor agonist, induces reflexive locomotion in decapitated flies. We show that the amount of locomotion induced changes as a function of the time of day, with the highest responsiveness to quinpirole during the subjective night. Furthermore, dopamine receptor responsiveness is under circadian control and depends on the normal function of the period gene. The head pacemaker is at least partly dispensable for the circadian modulation of quinpirole-induced locomotion, because changes in agonist responsiveness persist in decapitated flies that are aged for 12 h. This finding suggests a role for the period-dependent molecular oscillators in the body in the modulation of amine receptor responsiveness. PMID- 10677551 TI - Induction of topoisomerase I cleavage complexes by 1-beta -D arabinofuranosylcytosine (ara-C) in vitro and in ara-C-treated cells. AB - 1-beta-d-Arabinofuranosylcytosine (Ara-C) is a nucleoside analog commonly used in the treatment of leukemias. Ara-C inhibits DNA polymerases and can be incorporated into DNA. Its mechanism of cytotoxicity is not fully understood. Using oligonucleotides and purified human topoisomerase I (top1), we found a 4- to 6-fold enhancement of top1 cleavage complexes when ara-C was incorporated at the +1 position (immediately 3') relative to a unique top1 cleavage site. This enhancement was primarily due to a reversible inhibition of top1-mediated DNA religation. Because ara-C incorporation is known to alter base stacking and sugar puckering at the misincorporation site and at the neighboring base pairs, the observed inhibition of religation at the ara-C site suggests the importance of the alignment of the 5'-hydroxyl end for religation with the phosphate group of the top1 phosphotyrosine bond. This study also demonstrates that ara-C treatment and DNA incorporation trap top1 cleavage complexes in human leukemia cells. Finally, we report that camptothecin-resistant mouse P388/CPT45 cells with no detectable top1 are crossresistant to ara-C, which suggests that top1 poisoning is a potential mechanism for ara-C cytotoxicity. PMID- 10677552 TI - Ascorbic acid acts as an inhibitory transmitter in the hypothalamus to inhibit stimulated luteinizing hormone-releasing hormone release by scavenging nitric oxide. AB - Because ascorbic acid (AA) is concentrated in synaptic vesicles containing glutamic acid, we hypothesized that AA might act as a neurotransmitter. Because AA is an antioxidant, it might therefore inhibit nitric oxidergic (NOergic) activation of luteinizing hormone-releasing hormone (LH-RH) release from medial basal hypothalamic explants by chemically reducing NO. Cell membrane depolarization induced by increased potassium concentration [K(+)] increased medium concentrations of both AA and LH-RH. An inhibitor of NO synthase (NOS), N(G)-monomethyl-l-arginine (NMMA), prevented the increase in medium concentrations of AA and LH-RH induced by high [K(+)], suggesting that NO mediates release of both AA and LH-RH. Calcium-free medium blocked not only the increase in AA in the medium but also the release of LH-RH. Sodium nitroprusside, which releases NO, stimulated LH-RH release and decreased the concentration of AA in the incubation medium, presumably because the NO released oxidized AA to dehydro-AA. AA (10(-5) to 10(-3) M) had no effect on basal LH-RH release but completely blocked high [K(+)]- and nitroprusside-induced LH-RH release. N-Methyl d-aspartic acid (NMDA), which mimics the action of the excitatory amino acid neurotransmitter glutamic acid, releases LH-RH by releasing NO. AA (10(-5) to 10( 3) M) inhibited the LH-RH-releasing action of NMDA. AA may be an inhibitory neurotransmitter that blocks NOergic stimulation of LH-RH release by chemically reducing the NO released by the NOergic neurons. PMID- 10677553 TI - Isolation and characterization of powdery mildew-resistant Arabidopsis mutants. AB - A compatible interaction between a plant and a pathogen is the result of a complex interplay between many factors of both plant and pathogen origin. Our objective was to identify host factors involved in this interaction. These factors may include susceptibility factors required for pathogen growth, factors manipulated by the pathogen to inactivate or avoid host defenses, or negative regulators of defense responses. To this end, we identified 20 recessive Arabidopsis mutants that do not support normal growth of the powdery mildew pathogen, Erysiphe cichoracearum. Complementation analyses indicated that four loci, designated powdery mildew resistant 1-4 (pmr1-4), are defined by this collection. These mutants do not constitutively accumulate elevated levels of PR1 or PDF1.2 mRNA, indicating that resistance is not simply due to constitutive activation of the salicylic acid- or ethylene- and jasmonic acid-dependent defense pathways. Further Northern blot analyses revealed that some mutants accumulate higher levels of PR1 mRNA than wild type in response to infection by powdery mildew. To test the specificity of the resistance, the pmr mutants were challenged with other pathogens including Pseudomonas syringae, Peronospora parasitica, and Erysiphe orontii. Surprisingly, one mutant, pmr1, was susceptible to E. orontii, a very closely related powdery mildew, suggesting that a very specific resistance mechanism is operating in this case. Another mutant, pmr4, was resistant to P. parasitica, indicating that this resistance is more generalized. Thus, we have identified a novel collection of mutants affecting genes required for a compatible interaction between a plant and a biotrophic pathogen. PMID- 10677554 TI - UV light selectively coinduces supply pathways from primary metabolism and flavonoid secondary product formation in parsley. AB - The UV light-induced synthesis of UV-protective flavonoids diverts substantial amounts of substrates from primary metabolism into secondary product formation and thus causes major perturbations of the cellular homeostasis. Results from this study show that the mRNAs encoding representative enzymes from various supply pathways are coinduced in UV-irradiated parsley cells (Petroselinum crispum) with two mRNAs of flavonoid glycoside biosynthesis, encoding phenylalanine ammonia-lyase and chalcone synthase. Strong induction was observed for mRNAs encoding glucose 6-phosphate dehydrogenase (carbohydrate metabolism, providing substrates for the shikimate pathway), 3-deoxyarabinoheptulosonate 7 phosphate synthase (shikimate pathway, yielding phenylalanine), and acyl-CoA oxidase (fatty acid degradation, yielding acetyl-CoA), and moderate induction for an mRNA encoding S-adenosyl-homocysteine hydrolase (activated methyl cycle, yielding S-adenosyl-methionine for B-ring methylation). Ten arbitrarily selected mRNAs representing various unrelated metabolic activities remained unaffected. Comparative analysis of acyl-CoA oxidase and chalcone synthase with respect to mRNA expression modes and gene promoter structure and function revealed close similarities. These results indicate a fine-tuned regulatory network integrating those functionally related pathways of primary and secondary metabolism that are specifically required for protective adaptation to UV irradiation. Although the response of parsley cells to UV light is considerably broader than previously assumed, it contrasts greatly with the extensive metabolic reprogramming observed previously in elicitor-treated or fungus-infected cells. PMID- 10677555 TI - The production of recombinant proteins in transgenic barley grains. AB - The grain of the self-pollinating diploid barley species offers two modes of producing recombinant enzymes or other proteins. One uses the promoters of genes with aleurone-specific expression during germination and the signal peptide code for export of the protein into the endosperm. The other uses promoters of the structural genes for storage proteins deposited in the developing endosperm. Production of a protein-engineered thermotolerant (1, 3-1, 4)-beta-glucanase with the D hordein gene (Hor3-1) promoter during endosperm development was analyzed in transgenic plants with four different constructs. High expression of the enzyme and its activity in the endosperm of the mature grain required codon optimization to a C+G content of 63% and synthesis as a precursor with a signal peptide for transport through the endoplasmic reticulum and targeting into the storage vacuoles. Synthesis of the recombinant enzyme in the aleurone of germinating transgenic grain with an alpha-amylase promoter and the code for the export signal peptide yielded approximately 1 microgram small middle dotmg(-1) soluble protein, whereas 54 microgram small middle dotmg(-1) soluble protein was produced on average in the maturing grain of 10 transgenic lines with the vector containing the gene for the (1, 3-1, 4)-beta-glucanase under the control of the Hor3-1 promoter. PMID- 10677556 TI - The pollen determinant of self-incompatibility in Brassica campestris. AB - Many flowering plants possess self-incompatibility (SI) systems that prevent inbreeding. In Brassica, SI is controlled by a single polymorphic locus, the S locus. Two highly polymorphic S locus genes, SLG (S locus glycoprotein) and SRK (S receptor kinase), have been identified, both of which are expressed predominantly in the stigmatic papillar cell. We have shown recently that SRK is the determinant of the S haplotype specificity of the stigma. SRK is thought to serve as a receptor for a pollen ligand, which presumably is encoded by another polymorphic gene at the S locus. We previously have identified an S locus gene, SP11 (S locus protein 11), of the S(9) haplotype of Brassica campestris and proposed that it potentially encodes the pollen ligand. SP11 is a novel member of the PCP (pollen coat protein) family of proteins, some members of which have been shown to interact with SLG. In this work, we identified the SP11 gene from three additional S haplotypes and further characterized the gene. We found that (i) SP11 showed an S haplotype-specific sequence polymorphism; (ii) SP11 was located in the immediate flanking region of the SRK gene of the four S haplotypes examined; (iii) SP11 was expressed in the tapetum of the anther, a site consistent with sporophytic control of Brassica SI; and (iv) recombinant SP11 of the S(9) haplotype applied to papillar cells of S(9) stigmas, but not of S(8) stigmas, elicited SI response, resulting in inhibition of hydration of cross pollen. All these results taken together strongly suggest that SP11 is the pollen S determinant in SI. PMID- 10677557 TI - Elemental propagation of calcium signals in response-specific patterns determined by environmental stimulus strength. AB - Plant cells can respond qualitatively and quantitatively to a wide range of environmental signals. Ca(2+) is used as an intracellular signal for volume regulation in response to external osmotic changes. We show here that the spatiotemporal patterns of hypo-osmotically induced Ca(2+) signals vary dramatically with stimulus strength in embryonic cells of the marine alga Fucus. Biphasic or multiphasic Ca(2+) signals reflect Ca(2+) elevations in distinct cellular domains. These propagate via elemental Ca(2+) release in nuclear or peripheral regions that are rich in endoplasmic reticulum. Cell volume regulation specifically requires Ca(2+) elevation in apical peripheral regions, whereas an altered cell division rate occurs only in response to stimuli that cause Ca(2+) elevation in nuclear regions. PMID- 10677559 TI - Parsing executive processes: strategic vs. evaluative functions of the anterior cingulate cortex. AB - Event-related functional MRI and a version of the Stroop color naming task were used to test two conflicting theories of anterior cingulate cortex (ACC) function during executive processes of cognition. A response-related increase in ACC activity was present when strategic processes were less engaged, and conflict high, but not when strategic processes were engaged and conflict reduced. This is inconsistent with the widely held view that the ACC implements strategic processes to reduce cognitive conflicts, such as response competition. Instead, it suggests that the ACC serves an evaluative function, detecting cognitive states such as response competition, which may lead to poor performance, and representing the knowledge that strategic processes need to be engaged. PMID- 10677558 TI - The epidemiology of antibiotic resistance in hospitals: paradoxes and prescriptions. AB - A simple mathematical model of bacterial transmission within a hospital was used to study the effects of measures to control nosocomial transmission of bacteria and reduce antimicrobial resistance in nosocomial pathogens. The model predicts that: (i) Use of an antibiotic for which resistance is not yet present in a hospital will be positively associated at the individual level (odds ratio) with carriage of bacteria resistant to other antibiotics, but negatively associated at the population level (prevalence). Thus inferences from individual risk factors can yield misleading conclusions about the effect of antibiotic use on resistance to another antibiotic. (ii) Nonspecific interventions that reduce transmission of all bacteria within a hospital will disproportionately reduce the prevalence of colonization with resistant bacteria. (iii) Changes in the prevalence of resistance after a successful intervention will occur on a time scale of weeks to months, considerably faster than in community-acquired infections. Moreover, resistance can decline rapidly in a hospital even if it does not carry a fitness cost. The predictions of the model are compared with those of other models and published data. The implications for resistance control and study design are discussed, along with the limitations and assumptions of the model. PMID- 10677560 TI - Estrogen receptor variants are present in many normal human tissues. AB - Estrogen receptors (ER) are present in various reproductive tissues as well as in other tissues not considered classical targets for estrogen. A variety of alternatively spliced ER variants which co-exist with the wild-type receptor has also been described in many tissues. We analyzed the presence of both wild-type and variant receptors from normal human tissues of various origin using semi nested PCR and direct automated sequence of the amplified products. We demonstrate that many normal human tissues of various origin contain a number of spliced variants of the estrogen receptor that co-exist with the wild-type receptor. Variants lacking exons 2, 4, 5, and 7 are detected in a variety of normal human tissues. PMID- 10677561 TI - Familial neurohypophyseal diabetes insipidus associated with a novel mutation in the vasopressin-neurophysin II gene. AB - Familial neurohypophyseal diabetes insipidus (FNDI) is an autosomal dominant disorder of renal water conservation due to deficiency of arginine vasopressin as the result of mutations in the arginine vasopressin-neurophysin II (AVP-NPII) gene that encodes the hormone or its carrier protein. Thirty-one different mutations have been reported. In this study, we evaluated the AVP-NPII gene in a family with FNDI and identified a new mutation (1911Gright curved arrow A) in the coding sequence for NPII in affected family members. This mutation substitutes Tyr for 74 Cys in the NPII moiety. NPII is an intracellular carrier protein for AVP during the axonal transport from the hypothalamus to the posterior pituitary and contains 14 conserved cysteine residues forming 7 disulfide bonds. Because the mutation cosegregates with the phenotype, it is possible that this mutation causes neurohypophyseal diabetes insipidus in this family. PMID- 10677562 TI - Spectral karyotyping and chromosome banding studies of primary breast carcinomas and their lymph node metastases. AB - Three primary breast tumors and their lymph node metastases were characterized by G-banding, spectral karyotyping (SKY), and fluorescence in situ hybridization (FISH). In each case, the karyotypic abnormalities detected were similar in the primary tumor and its matched metastasis. Two of the pairs had near-diploid karyotypes with three to four chromosomal aberrations, whereas the third pair had a near-pentaploid chromosome content and many marker chromosomes in the primary tumor and a near-tetraploid chromosome number with almost the same marker chromosomes in the metastasis. SKY and FISH confirmed the karyotypic similarities between the primary tumors and their metastases and, in addition, improved the identification and characterization of marker chromosomes. One of the tumor pairs with near-diploid karyotypes had gain of 8q, 16q, and 17q, whereas the other had gain of 1q and chromosome 8 material in the form of ring chromosomes. The third pair had more complex chromosomal translocations and numerical changes resulting in net gain of material from chromosomes X, 1, 2, 6, 7, 14, 16, 19, and 20, and chromosome arms 8q and 11q, as well as net loss of material from chromosomes 3, 13, 18, 21, and 22. The present study underscores the need to combine conventional chromosome banding and molecular cytogenetic techniques in the cytogenetic analysis of solid tumors. PMID- 10677563 TI - Recombinant Semliki forest virus infects and kills human prostate cancer cell lines and prostatic duct epithelial cells ex vivo. AB - Semliki Forest Virus (SFV) is a broad host range RNA virus capable of high-level recombinant protein expression and apoptosis induction in many cell types. We have successfully used a recombinant, replication deficient SFV vector to express the LacZ marker gene product in seven human prostate cell lines, as well as in human prostate tissue explants. Flow cytometry revealed that 40-60% of PPC-1 prostate cancer cells died 24-72 h after infection with SFV-LacZ virus. Most human prostate cancer cell lines expressed high levels of recombinant protein. Infection of human prostate tissue ex vivo led to similarly high expression levels but the recombinant beta-galactosidase was confined to duct epithelial cells. Infection of cell and tissue cultures resulted in detachment of adherent cells from the culture surface and detachment of epithelial cells from the basement membrane of tissue. Our results indicate that SFV may be useful in targeting recombinant protein expression and apoptosis to prostatic duct epithelial cells. PMID- 10677564 TI - Induction of the putative protective protein ferritin by infrared radiation: implications in skin repair. AB - The modification of ferritin in human skin cells in vitro and in vivo following infrared-A irradiation by immunohistochemical analysis and ELISA were evaluated. In addition, we observed that IR-A is not capable of inducing frank damage to DNA (pyrimidine dimers, p53), induction of oxidative stress proteins (heme oxygenase, nitric oxide, superoxide dismutase, heat shock proteins) or proteases (collagenase, stromelysin, gelatinase) involved in carcinogenesis and photoaging of the skin. in vivo, basal levels of ferritin were heterogeneous for all individuals tested but all showed ferritin to stain precisely in the basal layer of unirradiated epidermis. Following IR-A radiation, the ferritin increase was localized to epidermal tissue and showed an increase from 120 to 220%. Parallel to the in vivo analysis, dermal fibroblasts were cultured from six individuals. Quantitative analysis for ferritin in cultured fibroblasts was assessed by ELISA and increases were seen to be dose-dependent and up to 130% of basal levels of ferritin following infrared-A. Our findings indicate that the putative defense system of ferritin that exists in human skin in vivo can be induced by infrared-A radiation and that these wavelengths may prove to be beneficial for human skin. Importantly, following the same doses of IR-A that induced ferritin levels, there was no alteration seen for nuclear DNA type damage, oxidative stress proteins or proteases involved in the degradation of skin. The increased concentrations of this antioxidant in human skin following acute UV radiation could afford increased protection against subsequent oxidative stress. PMID- 10677565 TI - Role of HIF-1 as a transcription factor involved in embryonic development, cancer progression and apoptosis (review). AB - Hypoxia-inducible factor-1 (HIF-1) is a transcription factor first identified as being activated by hypoxia but also in normoxic conditions by insulin and IGF-2. It is able to induce the expression of glycolytic genes and hence the ATP production, it also regulates the expression of the angiogenic factor VEGF and stimulates erythropoiesis via EPO production. HIF-1 is a protein necessary for the normal embryonic and cardiovascular system development, but seems to be also involved in cancer progression and apoptosis. Thus, it appears that HIF-1 plays a central role in normal cellular functions and in tissue metabolism but it is also involved in pathological evolutions raising its interest as a therapeutic target. In this review, we summarize the dual role of HIF-1 as a major component of the embryo development, as well as an element of tumor progression and of anoxia induced apoptosis. PMID- 10677566 TI - Suppressive effect of genistein on rat bone osteoclasts: involvement of protein kinase inhibition and protein tyrosine phosphatase activation. AB - The suppressive effect of genistein on osteoclast-like multinucleated cells from rat femoral tissues was investigated. The bone cells isolated from rat femoral tissues were cultured for 48 h in an alpha-minimal essential medium (5% fetal bovine serum) containing either vehicle or genistein (10(-7)-10(-5) M). Osteoclasts were estimated by staining for tartrate-resistant acid phosphatase, a marker enzyme of osteoclasts. The presence of genistein caused a significant decrease in the number of osteoclasts. Such a decrease was also seen in the presence of calcium choride (10(-5) M). Magnesium chloride (10(-5)-10(-3) M), a blocker of Ca2+ channels, had no effect on the number of osteoclasts. The effect of genistein (10(-5) M) or calcium (10(-3) M) in decreasing osteoclasts was significantly prevented by the presence of magnesium (10-3 M). Vanadate (10(-6) 10(-4) M), an inhibitor of protein tyrosine phosphatase activity, did not have an effect on the number of osteoclasts. The genistein's effect was not altered by vanadate. When isolated osteoclasts were cultured for 24 h in the presence of genistein (10(-7)-10(-5) M), protein kinase activity in the 5500 g supernatant of homogenate of the cells was significantly decreased, while protein tyrosine phosphatase activity was significantly elevated. Such an effect was also seen by the addition of genistein (10(-7)-10(-5) in the enzyme reaction mixture in vitro. The present study suggests that the suppressive effect of genistein on rat bone osteoclasts is partly involved in the inhibition of protein kinase and the activation of protein tyrosine phosphatase in osteoclasts. PMID- 10677567 TI - Molecular interactions between presenilin and calpain: inhibition of m-calpain protease activity by presenilin-1, 2 and cleavage of presenilin-1 by m-, mu calpain. AB - Several mutations of presenilin (PS)-1, 2 result in early onset Alzheimer disease. Using the yeast two-hybrid system, the interaction between PS2 loop domain and the C-terminal region of mu-calpain was previously identified. Calpain is a calcium dependent-protease and there are two isoforms, m-calpain and mu calpain, which differ in the calcium concentration required for activation. m Calpain needs about 10(-3) M calcium ions, whereas mu-calpain about 10(-5) M. When PS and calpain were separately expressed in COS cells by cDNA transfection and then combined in vitro, or both were co-transfected to be co-expressed in vivo in COS cells, PS1 and PS2 reduced the casein proteolysis activity of m calpain but not that of mu-calpain. Some of the PS mutations related to Alzheimer disease decreased this inhibitory activity. On the other hand, PS1 was cleaved by m-calpain and mu-calpain at a different site from those already reported (constitutive cleavage or alternative cleavage). These results suggest a regulatory function of presenilin on the calpain system. PMID- 10677568 TI - Regression of experimental liver tumor after distant intra-hepatic injection of cytosine deaminase-expressing tumor cells and 5-fluorocytosine treatment. AB - Cytosine deaminase (CD) gene of E. coli converts the non-toxic compound 5 fluorocytosine (5-FC) into 5-fluorouracil. We have introduced a vector expressing the CD gene in a rat colon carcinoma cell line. Expression of the CD gene confers 5-FC sensitivity to these cells in vitro and in vivo. In a bifocal model consisting in a simultaneous engrafment of a CD+ tumor on one lobe of the liver and a wild-type parental tumor on the opposite lobe, treatment with 5-FC results in regression of both type of tumors, indicating the existence of a distant bystander effect. PMID- 10677569 TI - Cancer cell selectivity of 5-iodo-6-aminobenzopyrone (INH2BP) and methyl-3,5 diiodo-4(4'-methoxyphenoxy) benzoate (DIME). AB - The cellular pharmacologic actions, as measured by cell killing, of INH2BP, DIME and INO2BA (+ BSO) were determined in three types of cancer cells and compared to their action on quiescent confluent human foreskin fibroblast (HSF) and pre confluent growing fibroblasts. The confluent HSF cells were completely refractory to the action of INH2BP and DIME, but were killed by INO2BA (+ BSO). Proliferating HSF and all three tumor cell types were killed by all three drugs. The apparent in vivo tumor specificity of INH2BP and DIME is explained by preferential cell cycle dependent selective drug uptake into tumor cells and by drug metabolism that reverses drug action in less vigorously cycling normal cells. The covalent binding of iodonitrosobenzamide (formed from INO2BA) and its toxicity are regulated by the concentration of GSH, and exhibit no cell cycle selectivity. PMID- 10677570 TI - Transcript heterogeneity of the human gene for Ca2+-binding protein regucalcin. AB - Regucalcin is a Ca2+-binding protein which plays a regulatory role in liver cell functions related to Ca2+. In this study we have cloned and characterized cDNA for regucalcin from human liver and human hepatoma cell line Hep G2 by screening and rapid amplification of cDNA ends (RACE). The nucleotide sequences of the clones revealed that they were identical in their coding region and differed only in their 5' untranslated regions (UTRs). Northern blot analysis showed that regucalcin mRNA in the Hep G2 was longer than that of the liver. The present study demonstrates the existence of transcript heterogeneity of the human gene for regucalcin. PMID- 10677571 TI - Immobilization of tumoral pancreatic islet cells on a two-dimensional microsupport. AB - A new procedure for the immobilization of tumoral pancreatic islet cells to a two dimensional microsupport is presented. Tumoral islet cells of the RINm5F line (0.7x10(6) cells/ml) were immobilized to two-dimensional microcarriers (16.6 cm2/ml). Within 24 h of culture, and as judged from the number of cells, their protein or insulin content, less than 10% of the cells escaped immobilization. The metabolic response of the immobilized cells to D-glucose was well preserved, the paired ratio between D-[U-14C] glucose oxidation and D-[5-3H]glucose utilization being even significantly higher in immobilized than free cells. The advantages of this novel approach in the perspective of islet cell transplantation are underlined. PMID- 10677572 TI - The H9/M8166 tropism of various HIV-1 mutants is determined by distinct cellular factors (review). AB - We have previously shown that a number of human immunodeficiency virus type 1 (HIV-1) mutants generated in vitro display a replication-defect in a cell dependent manner. Of the mutants of this category, those of gag, vif, and env mutants do not grow at all in lymphocytic H9 (non-permissive) cells, but quite well in M8166 (permissive) cells. To determine whether the cell-dependent growth of the mutants is functionally related to each other, a number of double mutants were constructed, and monitored for their replication and biochemical property in various cells. The results obtained have indicated that there are multiple cellular factors responsible for the growth phenotype. Together with our previous findings on the host cell-dependent mutants of HIV-1, it is concluded that number of distinct cellular factors are involved in the various steps in HIV-1 replication cycle. PMID- 10677573 TI - Construction of new amplifier expression vectors for high levels of IL-2 gene expression. AB - The success of IL-2 gene therapy in cancer is in part dependent on the development of high level IL-2 gene expression vectors. Currently, expression vectors based on the human cytomegalovirus (CMV) promoter give the highest levels of expression. We have attempted to construct new IL-2 expression vectors to test whether gene expression can be further increased. The first approach was to use the new SR-alpha promoter to control IL-2 gene expression. The second approach was to combine the Tat transcription activator gene and the HIV 1 and 2 promoters in the same construct so that the levels of gene expression can be amplified. Transient transfection results using the human colon cancer cell line SW480 showed that the SR-alpha promoter yields similar levels of activity as the CMV promoter. However, the HIV 1 and 2 promoter-based amplifier constructs produced 11 and 28 times more secreted IL-2 than the CMV promoter control. The augmented activity of the amplifier constructs was dependent on the presence of the Tat gene and the transcriptional units must be placed in the same orientation. Reducing the size of the vectors by elimination of the neomycin selectable marker did not increase the activity of the constructs. PMID- 10677574 TI - Molecular medicine: the present and the future. AB - In the recent past, major achievements have been obtained in the understanding of the molecular defects at the basis of several different diseases. The field of 'Molecular Medicine' has thus become more solid, and several reports have been published linking the basic molecular investigation to the clinical practice. In line with this new approach to medicine a Symposium was organized where the linkage between investigations in basic science could be explored in view of clinical disorders, and vice versa. in Rosario, Argentina, September 9-11, 1999, molecular biologists, molecular pathologists and clinicians discussed the molecular defects possibly at the basis of some common diseases. This report summarizes the presentations and discussions during the symposium. PMID- 10677575 TI - Therapeutically targeting lymphocyte energy metabolism by high-dose glucocorticoids. AB - Lymphocytes use a considerable amount of energy, mainly in the form of ATP, especially when they become stimulated following activation by antibodies or mitogens. Cellular respiration is the major energy source, and in quiescent cells the ATP produced is used to drive protein synthesis and sodium transport. In stimulated cells there is significantly higher ATP production to balance the higher ATP demand of specific processes resulting from activation. The major ATP consuming processes under these conditions are protein synthesis and Na(+),K(+) ATPase (about 20% each), while Ca(2+)-ATPase and RNA/DNA syntheses contribute about 10% each. There is a wealth of available information about glucocorticoid effects on lymphocytes, but here we focus on the extent to which this lymphocyte bioenergetic machinery is targeted by glucocorticoids when they are used therapeutically at high doses. High-dose glucocorticoids have been shown recently to interfere with processes that are essential for the activation and maintenance of lymphocytes, such as sodium and potassium transport. Therefore, in this article we describe the bioenergetics of lymphocytes in resting, activated, and glucocorticoid-treated states and present a concept for discussion to describe the relationship among these states in fundamental and clinical terms. PMID- 10677576 TI - Attenuation of interleukin-8 production by inhibiting nuclear factor-kappaB translocation using decoy oligonucleotides. AB - Interleukin-8 (IL-8), a monocyte-derived neutrophil chemoattractant factor, is a polymorphonuclear neutrophil chemotaxin that is involved in a number of inflammatory disorders. Transcription of the IL-8 gene is controlled by regulatory proteins, including nuclear factor-kappaB (NF-kappaB), a family of proteins that is important in the transcriptional control of a number of genes. When cells are activated, NF-kappaB translocates from the cytoplasm to the nucleus, where it activates transcription by binding to a specific sequence within the 5' untranslated region of the gene. During translocation, NF-kappaB is potentially susceptible to diversion by oligonucleotides that contain the binding sequence for this protein. In the current study, we produced phosphorothioate modified oligonucleotides containing the specific DNA sequence that NF-kappaB binds within the IL-8 gene. We then investigated the effects of transfection of monocytes with these oligonucleotides on interleukin-1beta (IL-1beta)-stimulated IL-8 production, IL-8 mRNA expression, and NF-kappaB binding activity. We found that transfection with these oligonucleotides significantly inhibited monocyte IL 8 production. A single-stranded oligonucleotide with two copies of the NF-kappaB binding sequence was the most potent of those tested. This single-stranded oligonucleotide also inhibited IL-1beta-induced translocation of NF-kappaB to the nucleus and reduced IL-8 mRNA expression. These studies demonstrated that monocyte production of IL-8 can be attenuated using a single-stranded oligonucleotide that binds a transcriptional activating protein before it translocates to the cell nucleus. This approach ultimately may be useful in the control of inflammation involved in a number of diseases. PMID- 10677577 TI - Uptake of the nitroimidazole drug megazol by African trypanosomes. AB - Megazol, CL 64,855 (2-amino-5-[1-methyl-5-nitro-2-imidazolyl]-1,3, 4-thiazole) has been shown to be extremely effective in clearing experimental infections of African trypanosomes. An unusual amino-purine transporter termed P2, implicated in the transport of both the diamidine and melaminophenyl arsenical classes of drug in Trypanosoma brucei, recognised chemical groups on compounds which are also present on megazol. Megazol interacted with this carrier protein, as judged by its ability to inhibit P2 adenosine transport and to abrogate in vitro arsenical-induced lysis in a dose-dependent manner. However, parasites resistant to melaminophenyl arsenical and diamidine drugs due to lack of the P2 transporter showed no resistance to megazol. This is because passive diffusion represented the major route of entry. Initial rates of uptake were not saturable within the limit of megazol's solubility and did not conform to thermodynamic precepts compatible with carrier-mediated uptake. Adenosine and other P2 transporter substrates, even at high concentration, had little impact on megazol uptake. Uptake was biphasic, with a very rapid equilibration across the membrane followed by a slower accumulation over time. The equilibration phase represented a simple passive diffusion, with the subsequent uptake probably being due to metabolism of the drug. PMID- 10677578 TI - Contribution of several metabolites of the vitamin D analog 20-epi-22-oxa 24a,26a,27a-tri-homo-1,25-(OH)(2) vitamin D(3) (KH 1060) to the overall biological activity of KH1060 by a shared mechanism of action. AB - The synthetic 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) analog 20-epi-22-oxa 24a,26a,27a-tri-homo-1,25-(OH)(2)vitamin D(3) (KH1060) is considerably more potent than its cognate hormone. The mechanism of action of KH1060 includes interaction with the vitamin D receptor (VDR). We previously showed that KH1060 increases VDR stability in ROS 17/2.8 osteoblastic cells by inducing a specific conformational change in the VDR. KH1060 is metabolized, both in vivo and in vitro, into several stable products. In the present study, we investigated whether these metabolites might contribute to the increased biological activity of KH1060. We found that the potencies of two of these metabolites, 24a-OH-KH1060 and 26-OH-KH1060, were similar to that of 1,25-(OH)(2)D(3) in inducing osteocalcin production by the osteoblast cell line ROS 17/2.8. This report further showed that these metabolites had the same effects as KH1060 on VDR: they increased VDR stability in ROS 17/2.8 cells, while limited proteolytic analysis revealed that they caused a conformational change in the VDR, resulting in an increased resistance against proteolytic cleavage. Furthermore, as shown in gel mobility shift assays, both compounds clearly induced VDR binding to vitamin D response elements. Together, these results show that the potent in vitro activity of KH1060 is not only directed by the effects on the VDR conformation/stabilization of the analog itself, but also by certain of its long lived metabolites, and emphasizes the importance of detailed knowledge of the metabolism of synthetic hormonal analogs. PMID- 10677579 TI - Camptothecin-stabilised topoisomerase I-DNA complexes in leukaemia cells visualised and quantified in situ by the TARDIS assay (trapped in agarose DNA immunostaining). AB - We have shown that the TARDIS assay (trapped in agarose DNA immunostaining) can be used to detect DNA-topoisomerase I (topo I) cleavable complexes in situ in individual cells following treatment with topo I-targeting drugs. This assay is a modification of the assay for DNA-topoisomerase II (topo II) cleavable complexes (Willmore et al., Mol Pharmacol 53: 78-85, 1998). Drug-stabilised topo I-DNA complexes were detected in situ by topo I-specific primary antibodies and then visualised using fluorescein isothiocyanate conjugated second antibodies. Immunofluorescence was then quantified using a cooled slow-scan coupled device camera and image analysis procedures. Camptothecin (CPT) was shown to stabilise topo I-DNA cleavable complexes in whole cells in a dose-dependent manner in both CCRF-CEM and K562 cells and in lymphoblasts from an adult with newly diagnosed acute myeloid leukaemia treated ex vivo with CPT. In K562 cells, cleavable complexes were found to be maximal between 30 and 90 minutes continuous exposure of CPT, and approximately 78% of cleavable complexes formed in these cells were found to be reversed within 5 minutes of drug removal. It has also been shown that the immunofluorescence detected by the TARDIS assay was specific for topo I targeting agents. Hence, the TARDIS assay provides a powerful tool to determine the levels of drug-stabilised cleavable complexes in whole cells and thereby aid in the understanding of the mechanism of interaction between topo I-targeting drugs and their target. PMID- 10677580 TI - A novel action of the antianginal drug bepredil: induction of internal Ca(2+) release and external Ca(2+) influx in Madin-Darby canine kidney (MDCK) epithelial cells. AB - The effect of the antianginal drug bepridil on Ca(2+) signaling in Madin-Darby canine kidney (MDCK) cells was investigated by using fura-2 as a Ca(2+) probe. Bepridil at 10-50 microM evoked a significant rise in cytosolic free Ca(2+) concentration ([Ca(2+)](i)) in a dose-dependent manner. The [Ca(2+)](i) rise consisted of an immediate initial rise and a slow decay. Removal of external Ca(2+) partly inhibited the Ca(2+) signals by reducing both the initial rise and the decay phase, suggesting that bepridil activated both external Ca(2+) influx and internal Ca(2+) release. In Ca(2+)-free medium, pretreatment with 50 microM bepridil nearly abolished the Ca(2+) release induced by thapsigargin (1 microM), an endoplasmic reticulum Ca(2+) pump inhibitor, and vice versa, pretreatment with thapsigargin inhibited most of the bepridil-induced Ca(2+) release, suggesting that the thapsigargin-sensitive Ca(2+) store was the main source of bepridil induced Ca(2+) release. Bepridil (50 microM) induced considerable Mn(2+) quench of fura-2 fluorescence at an excitation wavelength of 360 nm, which was partly inhibited by La(3+) (0.1 mM). Consistently, La(3+) (0.1 mM) pretreatment significantly inhibited the bepridil-induced [Ca(2+)](i) rise. Addition of 3 mM Ca(2+) induced a significant [Ca(2+)](i) rise after prior incubation with 10-50 microM bepridil in Ca(2+)-free medium, suggesting that bepridil induced dose dependent capacitative Ca(2+) entry. However, 50 microM bepridil inhibited 1 microM thapsigargin-induced capacitative Ca(2+) entry by 38%. Pretreatment with aristolochic acid (40 microM) so as to inhibit phospholipase A(2) inhibited 50 microM bepridil-induced internal Ca(2+) release by 42%, but inhibition of phospholipase C with U73122 (2 microM) or inhibition of phospholipase D with propranolol (0.1 mM) had little effect, suggesting that bepridil induced internal Ca(2+) release in an inositol 1,4,5-trisphosphate-independent manner that could be modulated by phospholipase A(2)-coupled events. This is the first report providing evidence that bepridil, currently used as an antianginal drug, induced a rise in [Ca(2+)](i) in a non-excitable cell line. PMID- 10677581 TI - Pharmacological properties of rat brain fatty acid amidohydrolase in different subcellular fractions using palmitoylethanolamide as substrate. AB - In the present study, the pharmacological properties of fatty acid amide hydrolase (FAAH) in subcellular fractions of rat brain were investigated using palmitoylethanolamide (PEA) and arachidonyl ethanolamide (anandamide, AEA) as substrates. FAAH hydrolysed [(3)H]PEA in crude homogenates with median K(m) and V(max) values of 2.9 microM and 2.14 nmol.(mg protein)(-1).min(-1), respectively. [(3)H]PEA hydrolysis was inhibited both by non-radioactive AEA (with a K(i) value very similar to the K(m) value for [(3)H]AEA as substrate using the same assay) and by R(-)ibuprofen (mixed-type inhibition K(i) and K'(i) values 88 and 720 microM, respectively). FAAH activity towards both [(3)H]PEA and [(3) myelin = cytosol, but there were no differences between the relative activities towards the two substrates in any of the fractions. [(3)H]PEA hydrolysis in mitochondrial, myelin, microsomal, and synaptosomal fractions was inhibited by oleyl trifluoromethylketone, phenylmethylsulphonyl fluoride, and the R(-)- and S(+)-enantiomers of the nonsteroidal anti-inflammatory drug ibuprofen, with mean IC(50) values in the ranges 0.028-0.041, 0.37-0.52, 67-110, and 130-260 microM, respectively. It is concluded that the pharmacological properties of FAAH in the different subcellular fractions are very similar. PMID- 10677582 TI - Biological activity of hexitol nucleic acids targeted at Ha-ras and intracellular adhesion molecule-1 mRNA. AB - Hexitol nucleic acid (HNA) is a new steric blocking oligonucleotide, hybridizing sequence selectively with RNA. The biological activity of HNA was evaluated in an in vitro translation arrest system targeting Ha-ras mRNA and in a cellular system targeting intracellular adhesion molecule-1 (ICAM-1) expression. HNA very efficiently and selectively inhibited Ha-ras mRNA translation (IC(50) of 50 nM) when targeted at the translation initiation region. When targeting at the 12th codon region, a gap-mer approach was needed to inhibit mRNA translation. Similarly, HNA inhibited ICAM-1 expression in keratinocytes when targeting at codon sequences. In this test system, HNA is less active but more selective than phosphorothioates, but needs lipofection to become active in keratinocytes. This new steric blocker may be an efficient antisense agent providing that enough material can be brought into cells. PMID- 10677583 TI - Effects of sodium deoxycholate and sodium caprate on the transport of epirubicin in human intestinal epithelial Caco-2 cell layers and everted gut sacs of rats. AB - The effects of sodium deoxycholate (Deo-Na), a bile salt, and sodium caprate (Cap Na), a fatty acid, on the transport of epirubicin were investigated in both the human colon adenocarcinoma (Caco-2) cell line and the everted gut sacs of the rat jejunum and ileum. The possible use of these two potent absorption enhancers as multidrug resistance (MDR) reversing agents also was examined. Epirubicin uptake experiments using a flow cytometer showed that Deo-Na and Cap-Na significantly increased the accumulation of epirubicin in Caco-2 cells. These two enhancers significantly increased apical to basolateral absorption of epirubicin across Caco-2 monolayers and mucosal to serosal absorption of epirubicin in the rat jejunum and ileum. Moreover, the addition of Deo-Na or Cap-Na significantly reduced the basolateral to apical efflux of epirubicin across Caco-2 monolayers. The co-presence of verapamil, one typical P-glycoprotein (P-gp) substrate, and Deo-Na or Cap-Na demonstrated further reduction of epirubicin efflux. The study suggests that inhibition of P-gp or other transporter proteins located in the intestines may be involved, at least partially, in the reduction of epirubicin efflux. In conclusion, the therapeutic efficacy of epirubicin may be improved by the use of such low toxicity excipients as absorption enhancers and MDR modulators in formulations. PMID- 10677584 TI - Effect of the beta-adrenoceptor agonist flerobuterol on serotonin synthesis in the rat brain. AB - The influence of 2- and 14-day treatments with flerobuterol, a preferential beta(2)-adrenoceptor agonist, on regional serotonin (5-HT) synthesis in the rat brain was studied by autoradiography using alpha-[(14)C]methyl-L-tryptophan. Flerobuterol was delivered at a rate of 0.5 mg/kg/day using osmotic pumps implanted s.c. The 2-day flerobuterol treatment significantly increased plasma Trp, both free and total, and decreased plasma Leu and Ile. This resulted in a significant increase in the facilitated transport of Trp. There was a significant increase in the synthesis of 5-HT in the 2-day treatment group in the dorsal and median raphe as well as in all postsynaptic structures, with the exception of the hypothalamus. In contrast, after a 14-day treatment, the enhanced facilitated transport of Trp was no longer present, and the increase in the rate of 5-HT synthesis persisted only in the parietal and occipital cortex and the superior colliculus. These data suggest that flerobuterol, similar to other beta adrenergic agonists, acutely increases 5-HT synthesis, in part, through an elevation of brain Trp availability. PMID- 10677585 TI - The decrease in total collagen fibers in the liver by hepatocyte growth factor after formation of cirrhosis induced by thioacetamide. AB - Liver cirrhosis is an inveterate disease accompanying fibrosis, hepatocyte damage, and liver dysfunction. In this study, the therapeutic effects of recombinant human hepatocyte growth factor (rhHGF) on liver cirrhosis were examined in rats administered thioacetamide (TAA). Repeated administration of TAA for 10 weeks to rats induced liver cirrhosis with collagen nodes and pseudo-lobe generation, a condition that was pathologically similar to that in humans. Administration of rhHGF after the formation of liver cirrhosis markedly decreased the incidence of pathological fibrosis and the degree of fibrosis as measured by a computed image analysis system. Continuous administration of rhHGF by infusion pump was more effective than bolus administration. Northern blotting analysis showed that rhHGF reduced mRNA levels of procollagen alpha2(I), alpha1(IV), and transforming growth factor-beta1 (TGF-beta1) that were stimulated in the TAA treated liver. The labeling index of hepatocytes increased following administration of rhHGF in this model. These observations suggest that the pathological recession of liver fibrosis is the result of the reduction of TGF beta1 and collagen synthesis and, in part, of the stimulation of mitosis of hepatocytes directly by rhHGF and indirectly by TGF-beta1 reduction in the cirrhotic liver. These results demonstrate the usefulness of rhHGF as a therapeutic agent in liver cirrhosis. PMID- 10677586 TI - Novel ring A stereoisomers of 2-methyl-1alpha,25-dihydroxyvitamin D(3) and 2 methyl-20-epi-1alpha,25-dihydroxyvitamin D(3): transactivation of target genes and modulation of differentiation in human promyelocytic leukemia (HL-60) cells. AB - We evaluated the biological activity of two sets of ring A stereoisomers of 2 methyl-1alpha,25-dihydroxyvitamin D(3) (2-methyl-1alpha,25(OH)(2)D(3)) and 2 methyl-20-epi-1alpha, 25-dihydroxyvitamin D(3) (2-methyl-20-epi 1alpha,25(OH)(2)D(3)) in terms of the following: transactivation of a rat 25 hydroxyvitamin D(3)-24-hydroxylase gene promoter including two vitamin D response elements (VDREs) and a human osteocalcin gene promoter including a VDRE in transfected human osteosarcoma (MG-63) cells; a vitamin D receptor (VDR)-mediated response using a VDR-GAL4 one-hybrid luciferase reporter system and a retinoid X receptor alpha (RXRalpha)-mediated response using an expressed VDR/RXRalpha-GAL4 modified two-hybrid luciferase reporter system in transfected human epitheloid carcinoma, cervix (HeLa) cells; and modulation of cell surface CD11b antigen expression in human leukemia (HL-60) cells. All the diastereomers of both analogues exhibited unique biological activity profiles depending upon the configurations of the C-1 and C-3 hydroxyl groups, the C-2 methyl group in ring A, and the C-20 methyl group in the side chain. Of the eight possible diastereomers of the 2-methyl analogues, 2alpha-methyl-1alpha,25(OH)(2)D(3) was the most potent and exhibited comparable or even greater biological potency than 1alpha,25(OH)(2)D(3). Of the eight possible diastereomers of the 2-methyl-20-epi analogues, 2alpha-methyl-20-epi-1alpha,25(OH)(2)D(3) was the most potent and exhibited 100- to 200-fold higher transcriptional potencies than 1alpha,25(OH)(2)D(3) and exceptionally high cell regulatory activities. 2beta methyl-20-epi-1alpha,25(OH)(2)D(3) was nearly as potent as its 2-epimer, 2alpha methyl-20-epi-1alpha,25(OH)(2)D(3), whereas its 20-epimer, 2beta-methyl 1alpha,25(OH)(2)D(3), was almost completely biologically inactive. In these respects, it can be postulated that the double modification of 2-methyl substitution and 20-epimerization to 1alpha,25(OH)(2)D(3) induces remarkable changes in a VDR/RXRalpha/VDRE-mediated signaling response and greatly enhances biological activity. The other striking finding was that 2beta-methyl-20-epi-3 epi-1beta,25(OH)(2)D(3) is transcriptionally more active than 1alpha,25(OH)(2)D(3) despite lacking the 1alpha-hydroxyl group, which was believed to be essential for expressing VDR-mediated gene transcription. Since the C-20 natural counterpart, 2beta-methyl-3-epi-1beta,25(OH)(2)D(3), was almost completely biologically inactive, 20-epimerization is probably responsible for activation of gene expression. Although earlier extensive structure-activity studies of vitamin D analogues showed stereochemistry at the C-1, C-3, and C-20 of 1alpha,25(OH)(2)D(3) to be the key structural motif for vitamin D action, our results clearly demonstrated that stereochemistry at the C-2 is also an important structural motif for vitamin D action and imply that 2-methyl substitution possibly induces conformational changes in ring A depending upon the combinations of configurations of the C-1 and C-3 hydroxyl groups with C-20 stereochemistry. Consequently, several of these analogues exhibit exceptionally high or unexpected biological activities at the molecular and cellular levels. These results suggest that 2-methyl substitution together with alterations of stereochemistry in both ring A and the side chain of 1alpha, 25(OH)(2)D(3) will provide useful analogues for structure-activity studies and development of therapeutic agents with unique biological activity profiles. PMID- 10677587 TI - Comparison of oxidative stress response parameters in newborn mouse liver versus simian virus 40 (SV40)-transformed hepatocyte cell lines. AB - Induction of approximately one dozen genes and/or enzyme activities in liver of the untreated newborn c(14CoS)/c(14CoS) mouse-when compared with the c(ch)/c(14CoS) heterozygote or the c(ch)/c(ch) wild-type-is the result of enhanced levels of reactive oxygenated metabolites originating from a block in the tyrosine degradation pathway. Oxidative stress activates genes via the electrophile response element, whereas dioxin activates genes via the receptor mediated aromatic hydrocarbon response element. Here, we compared several parameters in 14CoS/14CoS versus ch/ch newborn mouse liver with that in simian virus 40 (SV40)-transformed hepatocyte lines that had been derived from newborn liver. We showed in this study that: (a) NADP(H):quinone oxidoreductase and UDP glucuronosyltransferase 1A6 mRNA levels were increased in both the (untreated) 14CoS/14CoS newborn liver and cell line; (b) aldehyde dehydrogenase 3A1 mRNA was increased by both oxidative stress and dioxin in hepatocyte cultures, but was not detectable in liver of the intact mouse; (c) the glutathione S-transferase GSTA1, GSTP1, GSTA3, and GSTM1 mRNA levels were increased by oxidative stress in 14CoS/14CoS newborn liver, but these transcripts were either low or undetectable in the cell lines; (d) GSTA1 mRNA was up-regulated by the absence of cytochrome P450 1A1 (CYP1A1) activity (i.e. the Gsta1 gene is a member of the aromatic hydrocarbon [Ah] battery); and (e) GSTP1 mRNA was not up-regulated by the absence of CYP1A1 activity (i. e. Gstp1 is not a member of the [Ah] battery). The 14CoS/14CoS and ch/ch hepatocyte established cell lines were transformed with SV40, which expresses large T antigen; this gene product is known to bind to, and interact with, several cell cycle regulatory proteins such as p53 and the retinoblastoma protein-E2F complex. It is therefore likely that differences in the oxidative stress responses between the 14CoS/14CoS newborn liver and the immortalized hepatocyte cell line might be explained by the presence of large T antigen in the established cell line. PMID- 10677588 TI - Expression of nitric oxide-sensitive guanylyl cyclase subunits in human corpus cavernosum. AB - The muscles of the corpus cavernosum of the penis relax in response to stimulation of non-adrenergic, non-cholinergic nerves or nitric oxide (NO) donating drugs to elicit erection. It is generally assumed that NO mediates this effect via activation of soluble guanylyl cyclase and a subsequent increase in cyclic guanosine 3', 5'-monophosphate concentration. However, there are no data on the expression of this enzyme in human corpus cavernosum. The purpose of the present study was the molecular characterization of NO-sensitive guanylyl cyclase in human corpus cavernosum. RNA was extracted from tissue samples obtained from seven patients undergoing penile prosthetic surgery or correction of penile deviation. Reverse transcriptase-polymerase chain reaction (RT-PCR) with specific primers for the subunits of NO-sensitive guanylyl cyclase was performed, and PCR products were subcloned and sequenced. Specific amplification products encoding the alpha(1), beta(1), alpha(2), and beta(2) subunits were detected. In addition, we isolated a transcript encoding a novel variant beta(2) subunit. To test whether this novel transcript arises by alternative splicing or whether it is encoded by a separate gene, a 4000-bp clone of the corresponding genomic DNA sequence was isolated. Sequence analysis suggests that the novel beta(2) variant arises by alternative splicing from the same gene as the beta(2) subunit on chromosome 13. In conclusion, our findings suggest the presence of different subunit mRNAs of NO-sensitive guanylyl cyclase in human corpus cavernosum. PMID- 10677589 TI - Inhibitory effects of aclarubicin on nitric oxide production in aortic smooth muscle cells and macrophages. AB - The effects of aclaruhicin (ACR), an anthracycline antibiotic, on inducible nitric oxide (NO) synthesis was investigated in rat aortic smooth muscle cells (RASMCs) and RAW macrophages. ACR at concentrations as low as 0.1 microM significantly inhibited NO production induced by interleukin-1beta in RASMCs. About 5- to 10-fold higher concentrations of ACR were required for inhibition of interferon-gamma and lipopolipopolysaccharide-induced NO production in RAW cells. When ACR was subsequently administered to inducible NO synthase (iNOS) induction, the NO production was barely suppressed in RASMCs. Moreover, ACR (up to 10 microM) lacked direct inhibitory effects on iNOS activity in homogenates of these cells. ACR (0.1 microM) inhibited the expression of iNOS protein and mRNA in RASMCs without concomitant cytotoxic effects. ACR (>0.5 microM)-induced inhibition of NO production in RAW cells was associated with substantial cytotoxic effects as shown by measurement of lactate dehydrogenase release. These results suggest that ACR is a potent inhibitor of iNOS induction in vascular smooth muscle, but inhibits iNOS induction in macrophage only at high cytotoxic PMID- 10677590 TI - Differential expression of renal adenosine A(1) receptors induced by acute renal failure. AB - The distribution of renal adenosine A(1) receptors was investigated in rats with glycerol- or mercuric chloride (HgCl(2))-induced acute renal failure. Receptors were localised by autoradiography using [(3)H]8-cyclopentyl-1,3-dipropylxanthine ([(3)H]DPCPX), a selective A(1) adenosine receptor antagonist. In saline-injected control animals, significant labelling with [(3)H]DPCPX was detected in glomeruli, the inner stripe of outer medulla, and the inner medulla. Sixteen hours following induction of glycerol-induced acute renal failure (ARF), a 34% increase in labelling in glomeruli was noted compared to saline-injected controls, and by 48 hr, glomerular labelling had increased by 200%. In addition, 48 hr following glycerol injection, significant labelling was now detected in the cortical labyrinth and medullary rays whilst, in the inner medulla, labelling had decreased by 34%. By contrast to glycerol-induced ARF, the only significant change noted 48 hr following induction of HgCl(2)-induced ARF was a 39% decrease in labelling in the inner medulla. It is concluded that glycerol-induced ARF results in differential expression of renal adenosine A(1) receptors with increased expression in the cortex and reduced expression in the inner medulla. Increased density of A(1) receptors in glomeruli may account, at least in part, for the increased renal vasoconstrictor response to adenosine and depressed glomerular filtration rate noted previously in this type of acute renal failure. PMID- 10677591 TI - Involvement of hippocampal PKCbetaI isoform in the early phase of memory formation of an inhibitory avoidance learning. AB - Several evidences demonstrate that protein kinase C (PKC) is involved in hippocampal long-term potentiation (LTP) and in different forms of learning, including inhibitory avoidance training in rats. Here, we evaluated the levels of conventional PKC isozymes (alpha, betaI, betaII, gamma) in synaptic plasma membrane (SPM) fractions isolated from hippocampus of rats subjected to a one trial inhibitory avoidance paradigm. At 0, 30 and 120 min after training, there was a significant increase in the total amount of PKCbetaI. Densitometric analysis of the immunoblots showed an increase of 142+/-11% at 0 min, 193+/-16% at 30 min and 156+/-6% at 120 min after training relative to shocked control values. No changes were found in PKCbetaI levels in SPM fractions of the shocked animals relative to naive control values. No training-specific increments in the levels of PKCalpha, betaII and gamma were observed at any time point tested. However, an increase in PKCgamma levels was found in trained and shocked animals sacrificed 120 min after each experimental procedure. In addition, bilateral microinjections of a fairly selective inhibitor of PKCbetaI isozyme into the CA1 of the dorsal hippocampus produced amnesia when given 10 min before training, or 50, 110, but not 170 min, after training. Thus, the present findings demonstrate the participation of PKCbetaI in the early synaptic events responsible for the acquisition and consolidation of an inhibitory avoidance learning, and suggest a putative role of this presynaptic isozyme on the enhanced PKC-dependent B-50/GAP 43 phosphorylation previously detected by us during this associative learning. PMID- 10677592 TI - The influence of endogenous dopamine levels on the density of [3H]SCH23390 binding sites in the brain of the honey bee, Apis mellifera L. AB - This paper examines the relationship between endogenous dopamine (DA) levels and the density of [3H]SCH23390-binding sites in the brain of the adult worker honey bee. DA levels were reduced pharmacologically using a single 10 microl injection of either alpha-methyl-DL-p-tyrosine (AMT; 250 microg or 500 microg) or alpha methyl-DL-tryptophan (AMTP; 250 or 500 microg) into the haemolymph of the bee. In all cases, maximum depletion of DA was observed 3 h after treatment, but in bees treated with AMTP (250 or 500 microg) or with 250 microg AMT, DA levels returned to normal within 24 h of treatment. Neither AMT nor AMTP was selective for DA: both drugs also reduced serotonin (5-hydroxytryptamine, 5HT) levels in the brain. However, AMTP was more effective than AMT at depleting 5HT, whereas for DA, the reverse was true. Depletion of DA levels, using 250 microg AMT, led to a dramatic decline in the levels of specific binding of [3H]SCH23390, defined in this study as binding in the presence of 5x10(-6) M cis-(Z)-flupentixol (see Ref. [28] ). In contrast, naturally occurring diel fluctuations in DA levels, identified in the optic lobes of the brain, and changes in brain DA levels resulting from queenlessness, had no significant effect on the density of [3H]SCH23390-binding sites in the brain of the bee. Overall, these results indicate that under normal physiological conditions, there is no direct link in honey bees between changes in endogenous brain DA levels and the density of D(1)-like receptors labelled by [3H]SCH23390. PMID- 10677593 TI - Are neocortical gamma waves related to consciousness? AB - Previous research has shown that neocortical gamma waves (approximately 30-80 Hz) are continuously present during low voltage fast neocortical activity (LVFA) occurring during waking or active sleep. Gamma waves occur in a burst-suppression pattern in association with large amplitude slow waves during quiet sleep or anesthesia. The present experiments show that continuous gamma activity is also present in rats during LVFA occurring during surgical anesthesia (with ether, isoflurane or urethane) and that a burst-suppression pattern of gamma activity occurs during large amplitude slow waves occurring in the waking state either spontaneously in undrugged rats or as a result of treatment with parachlorophenylalanine and scopolamine. The amplitude of gamma activity occurring during anesthesia is variable but is often greater than it is in the normal waking state. It is concluded that the pattern of neocortical gamma wave activity is strongly related to the presence or absence of large amplitude slow waves but is quite independent of the state of behavioral arousal. Whether or not gamma wave activity is related to subjective awareness is a very difficult question which cannot be answered with certainty at the present time. PMID- 10677594 TI - N-type calcium channel blockers - tools for modulation of cerebral functional units? AB - According to in vitro and in vivo studies, the direct application of N-type calcium channel blockers as for instance omega-conotoxin GVIA (omega-ctx) potently inhibits the release of neurotransmitters like dopamine. To find out whether this effect could be used for modulation of neurological functions, omega ctx was used for continuous infusion into the functionally well characterized rat striatum. Over the 2-week time course of intrastriatal application, rats developed a decrease in spontaneous motor activity, spontaneous rotational asymmetry towards the side of application, and behavioral supersensitivity to apomorphine. After the end of infusion period, all functional deficits showed reversibility. The pattern of spontaneous neurological deficits - in particular supersensitivity to apomorphine - points to a substantial unilateral alteration of dopaminergic transmission due to omega-ctx, which is suggested also by an increase in dopamine receptor protein expression within the ipsilateral striatum. Time course and reversibility of neurological deficits caused by omega-ctx, as well as a lack of dopamine depletion contrast findings after selective destruction of dopaminergic neurons and support a functional modulation of dopaminergic transmission. The present study suggests that omega-ctx is an effective potent tool for the unilateral and reversible intracerebral modulation of neuronal circuits. Intracerebral application of omega-ctx could possibly open the way to therapeutic interventions. PMID- 10677595 TI - Cytochrome P-450 activities in human and rat brain microsomes. AB - The role of cytochrome P450 in the metabolism of dextromethorphan, amitriptyline, midazolam, S-mephenytoin, citalopram, fluoxetine and sertraline was investigated in rat and human brain microsomes. Depending on the parameters, the limit of quantification using gas chromatography-mass spectrometry methods was between 1.6 and 20 pmol per incubation, which generally contained 1500 microg protein. Amitriptyline was shown to be demethylated to nortriptyline by both rat and human microsomes. Inhibition studies using ketoconazole, furafylline, sulfaphenazole, omeprazole and quinidine suggested that CYP3A4 is the isoform responsible for this reaction whereas CYP1A2, CYP2C9, CYP2C19 and CYP2D6 do not seem to be involved. This result was confirmed by using a monoclonal antibody against CYP3A4. Dextromethorphan was metabolized to dextrorphan in rat brain microsomes and was inhibited by quinidine and by a polyclonal antibody against CYP2D6. Only the addition of exogenous reductase allowed the measurement of this activity in human brain microsomes. Metabolites of the other substrates could not be detected, possibly due to an insufficiently sensitive method. It is concluded that cytochrome P450 activity in the brain is very low, but that psychotropic drugs could undergo a local cerebral metabolism which could have pharmacological and/or toxicological consequences. PMID- 10677596 TI - Possible involvement of cytosolic phospholipase A(2) in cell death induced by 1 methyl-4-phenylpyridinium ion, a dopaminergic neurotoxin, in GH3 cells. AB - Previously we reported that 1-methyl-4-phenylpyridinium ion (MPP(+)), a dopaminergic neurotoxin, induced apoptosis of GH3 cells established from rat anterior pituitary. In the present study, the role of MPP(+) along with that of other apoptotic factors such as Ca(2+) and H(2)O(2) in cell death was examined. Ionomycin induced DNA fragmentation and lactate dehydrogenase (LDH) leakage in GH3 cells. H(2)O(2) also induced LDH leakage. Co-addition of MPP(+), in conditions where MPP(+) had no effect by itself, enhanced ionomycin- and H(2)O(2) induced cell death. Because the stimulation of phospholipase A(2) (PLA(2)) causing arachidonic acid (AA) release has been proposed to be involved in neuronal cell death, the effect of MPP(+) on AA release in GH3 cells was investigated. MPP(+) treatment for 8 h enhanced ionomycin- and H(2)O(2) stimulated AA release mediated by activation of cytosolic PLA(2) in a concentration-dependent manner, although MPP(+) by itself had no effect on AA release. An inhibitor of cytosolic PLA(2) inhibited MPP(+)-induced cell death. These findings suggest a synergistic effect of MPP(+) on Ca(2+)- and H(2)O(2) induced cell death, and the involvement of cytosolic PLA(2) activation in MPP(+) induced cell death in GH3 cells. Pretreatment with a caspase inhibitor or EGF did not modify the ionomycin- or H(2)O(2)-induced AA release, or enhancement by MPP(+), but the pretreatment inhibited the cell death in the presence and absence of MPP(+). The involvement of caspase(s) on activation of PLA(2) by MPP(+) was excluded, and EGF inhibited MPP(+)-induced cell death downstream of the AA release. PMID- 10677597 TI - Development of kindling and spontaneous seizures after massed stimulation of different loci in the rat piriform cortex. AB - Massed electrical stimulation of the anterior piriform cortex (PC) in rats using short (5 min) interstimulus intervals has previously been reported to induce severe chronic epilepsy with spontaneous seizures and has thus proposed to represent a novel model of temporal lobe epilepsy. In the present study, we used this stimulation protocol to evaluate the frequency and severity of recurrent spontaneous seizures produced in this way. In addition to the locus in the anterior PC previously used for massed stimulation (MS), we also stimulated rats via a locus in the transition zone between anterior and posterior PC ("central PC"), which previously was found to be more sensitive to electrical stimulation than various other loci in the anterior or posterior PC. During MS (71 stimulations for 1 s each at twice afterdischarge threshold), focal and infrequent secondary generalized seizures occurred in both groups, but there was no consistent progressive increase in seizure severity with increasing number of seizures, possibly as a result of postictal inhibitory processes. Following MS, rats were restimulated after 1, 2, 4, and 7 weeks, using five stimuli at 5-min interstimulus periods at each retest period. In both PC-implanted groups, seizure severity and seizure duration progressively increased over the period of the retests, indicating a delayed development of kindling. Spontaneous seizures were only observed rarely, so that MS of the PC is certainly no effective means of producing recurrent spontaneous seizures. PMID- 10677598 TI - The involvement of dopamine in nociception: the role of D(1) and D(2) receptors in the dorsolateral striatum. AB - Determination of the neuroanatomical and neurochemical factors that contribute to nociception is an essential element in the study and treatment of pain. Several lines of evidence have implicated nuclei and neurotransmitters within the basal ganglia in nociception. For example, previous studies have shown that dopamine receptors in the striatum are involved in acute nociception, however, it remains to be determined if dopamine receptors in the dorsolateral striatum are involved in persistent nociception. The purpose of the present study was therefore to determine whether activation or antagonism of dopamine receptors in the dorsolateral striatum influences the nociceptive responses of rats in the formalin test, a model of persistent pain. It was found that micro-injection of the non-selective dopamine antagonist haloperidol into the dorsolateral striatum increases formalin-induced nociception whereas injection of the non-selective dopamine agonist apomorphine reduces formalin-induced nociception. Injection of the D(1) antagonist SCH23390 or the D(1) agonist SKF38393 does not affect formalin-induced nociception. In contrast, injection of the D(2) antagonist eticlopride enhances formalin-induced nociception, whereas injection of the D(2) agonist quinpirole reduces formalin-induced nociception. These results provide additional evidence that dopamine receptors in the striatum are involved in nociception. Furthermore, this study strongly suggests that D(2), but not D(1), dopamine receptors in the dorsolateral striatum are involved in modulation of persistent nociception. PMID- 10677599 TI - N-Ethylmaleimide modulation of tetrodotoxin-sensitive and tetrodotoxin-resistant sodium channels in rat dorsal root ganglion neurons. AB - The effects of N-ethylmaleimide (NEM), an alkylating reagent to protein sulfhydryl groups, on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) sodium channels in rat dorsal root ganglion (DRG) neurons were studied using the whole cell configuration of patch-clamp technique. When currents were evoked by step depolarizations to 0 mV from a holding potential of -80 mV NEM decreased the amplitude of TTX-S sodium current, but exerted little or no effect on that of TTX-R sodium current. The inhibitory effect of NEM on TTX-S sodium channel was mainly due to the shift of the steady-state inactivation curve in the hyperpolarizing direction. NEM did not affect the voltage-dependence of the activation of TTX-S sodium channel. The steady-state inactivation curve for TTX-R sodium channel was shifted by NEM in the hyperpolarizing direction as that for TTX-S sodium channel. NEM caused a change in the voltage-dependence of the activation of TTX-R sodium channel unlike TTX-S sodium channel. After NEM treatment, the amplitudes of TTX-R sodium currents at test voltages below -10 mV were increased, but those at more positive voltages were not affected. This was explained by the shift in the conductance-voltage curve for TTX-R sodium channels in the hyperpolarizing direction after NEM treatment. PMID- 10677600 TI - Participation of Rel/NF-kappaB transcription factors in long-term memory in the crab Chasmagnathus. AB - The induction of gene expression has been correlated with long-lasting neuronal plasticity and long-term memory (LTM) formation. The fast activation of constitutive transcription factors by signaling mechanisms is thought to be the link between synaptic events and gene expression. However, only one constitutive transcription factor, CREB, has been shown to play a key role in several memory paradigms, both in vertebrates and invertebrates. Here, we report evidences for Rel/NFkappa-B constitutive transcription factors participation in memory. Using the LTM paradigm in the crab Chasmagnathus, an enhancement of NFkappa-B DNA binding activity was found after spaced training, which induces LTM, but not after massed training which yields an intermediate-term memory (ITM). Such finding is correlated with the requirement of protein synthesis for LTM consolidation but not for ITM. Furthermore, NFkappa-B activation was observed after 15 or 30 training trials, which are sufficient to induce LTM, but not after 5 or 10 trials, a number of trials insufficient to induce LTM. The kinetics of activation was studied and two waves of DNA-binding activity were found, similar to the time course described in other systems. NFkappa-B activation after training was also found in synaptosomal extracts. The latter result supports the hypothesis of a novel synapse-to-nucleus signaling system, in which the transcription factor is locally activated by synaptic events and then transported to the nucleus. PMID- 10677601 TI - Effects of nitric oxide gas on cat carotid body chemosensory response to hypoxia. AB - It has been proposed that nitric oxide (NO) is an inhibitory modulator of carotid body (CB) chemoreception to hypoxia. However, the effects of NO gas on carotid chemoreception have not been tested yet. The role played by NO has been revealed by the use of pharmacological tools (i.e., NO donors and NO synthase inhibitors). Here, we studied the effects of NO gas (25 ppm in N(2)) on the chemosensory response to hypoxia (PO(2) approximately 30 Torr) in the cat CB perfused in vitro. During steady hypoxic chemoreceptor excitation, bolus injections or perfusion of Tyrode equilibrated with NO reduced the increased frequency of carotid chemosensory discharges (f(x)). Perfusion for 2 min of Tyrode equilibrated with NO also reduced the rate of the rise of the chemosensory response, as well as the maximal amplitude, as compared with the normal chemosensory response to hypoxia. Present results provide direct evidence that NO gas is an inhibitory modulator of CB hypoxic chemoreception. PMID- 10677602 TI - Immunohistochemical detection of heme oxygenase-2 in the periodontal Ruffini ending of the rat incisor. AB - The present study was carried out to examine the occurrence of heme oxygenase-2 (HO-2) in the periodontal ligament of the rat incisor. HO-2-like immunoreactive ( IR) structures showed dendritic profiles, resembling the Ruffini endings, in the alveolar half of the ligament of rat incisor. Neither thin nerve fibers nor perivascular nerve fibers displayed HO-2-like immunoreactivity (-LI). No non neural elements exhibited HO-2-LI. Electron microscopy revealed that immunoreactions were diffusely observed in the axon terminals of the Ruffini endings, but neither terminal Schwann cells nor Schwann sheaths contained immunoreactions for HO-2. Both most neurons in the trigeminal ganglion and trigeminal mesencephalic nucleus showed HO-2-LI. The presence of HO-2 in the periodontal Ruffini endings and its absence in the periodontal thin nerve fibers suggest the involvement of carbon monoxide produced by HO-2 in mechanoreception in the periodontal ligament. PMID- 10677603 TI - Inhibition of L-homocysteic acid and buthionine sulphoximine-mediated neurotoxicity in rat embryonic neuronal cultures with alpha-lipoic acid enantiomers. AB - In the present report, we have set out to investigate the potential capacity of both the oxidised and reduced forms of RS-alpha-lipoic acid, and its separate R (+) and S-(-)enantiomers, to prevent cell death induced with L-homocysteic acid (L-HCA) and buthionine sulphoximine (BSO) in rat primary cortical and hippocampal neurons. L-HCA induced a concentration-dependent neurotoxic effect, estimated by cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction, in primary neurons, but was significantly more toxic for hippocampal (EC(50)=197 microM) compared with cortical neurons (EC(50)=1016 microM) whereas D HCA demonstrated only moderate (<20%) toxicity. On the other hand, cortical and hippocampal cultures were equally susceptible (341 and 326 microM, respectively) to the neurotoxic action of BSO. Antioxidants including butylated hydroxyanisole, propyl gallate and vitamin E protected cells against the neurotoxic effect of L HCA and BSO. However, N-acetyl-cysteine and tert-butylphenyl nitrone, although capable of abrogating L-HCA-mediated cell death showed no protective effect against BSO-mediated toxicity. RS-alpha-lipoic acid, RS-alpha-dihydrolipoic acid and the enantiomers R-alpha-lipoic acid and S-alpha-lipoic acid protected cells against L-HCA-mediated toxicity with EC(50) values between 3.1-8.3 microM in primary hippocampal neurons and 2.6-16.8 microM for cortical neurons. However, RS alpha-lipoic acid, RS-alpha-dihydrolipoic acid, and S-alpha-lipoic acid failed to protect cells against the degeneration induced by prolonged exposure to BSO, whereas the natural form, R-alpha-lipoic, was partially active under the same conditions. The present results indicate a unique sensitivity of hippocampal neurons to the effect of L-HCA-mediated toxicity, and suggest that RS-alpha lipoic acid, and in particular the R-alpha-enantiomeric form is capable of preventing oxidative stress-mediated neuronal cell death in primary cell culture. PMID- 10677604 TI - The effects of N-methyl-N-nitrosourea and azoxymethane on focal cerebral infarction and the expression of p53, p21 proteins. AB - If the activity of pro-apoptotic genes can be down-regulated by certain chemicals, cells may be protected from apoptosis. To test this hypothesis in a cerebral infarction model, we used N-methyl-N-nitrosourea (MNU) and azoxymethane (AOM), which were approved gene-modulating chemicals. A focal cerebral infarction was created by coagulation of the right middle cerebral artery and ipsilateral common carotid artery (CCA) and simultaneous transient occlusion of the contralateral CCA for 30 min in 25 adult Sprague-Dawley rats that were sacrificed 24 h later. In one group (n=7), MNU (5 mg/kg) was injected intravenously 30 min before initiation of ischemia. In another group (n=7), AOM (15 mg/kg) was administered intraperitoneally before 24 h of ischemia. The infarction volumes were checked and the brains were stained for p53 and p21 proteins. The width in micrometers of the peri-infarct area containing p53 or p21 protein-positive cells, and the number of p53 or p21 protein-positive cells (cells/HPF) were measured at an adjacent peri-infarct area. The AOM-treated group showed a significantly reduced infarction volume (by 42.5%, p<0.001), a significantly greater number of p53 positive cells (by 12.0%, p<0. 05), and a significantly wider p53 protein-positive area (by 15.6%, p<0.01) than the untreated group. AOM did not show any influence on the expression pattern of the p21 protein. MNU had no effect in the expression of p53 or p21 proteins. As a result, we concluded that AOM revealed a protective effect in ischemia by suppressing the pro apoptotic activity of the p53 gene. Safer chemicals that can modulate apoptotic genes, if any, will provide a new therapeutic modality for cerebral infarction. PMID- 10677605 TI - N-methyl-D-aspartate-type glutamate receptors are found in post-synaptic targets of adrenergic terminals in the thoracic spinal cord. AB - Adrenergic (C1) neurons in the rostral ventrolateral medulla (RVL) are sympathoexcitatory and project directly to sympathetic preganglionic neurons (SPNs) in the thoracic spinal cord. C1 neurons also contain glutamate which may mediate the excitatory effects of RVL stimulation on SPNs through the N-methyl-D aspartate (NMDA)-type receptor. Dual-labeling immunocytochemistry, combined with electron microscopy, was used to determine if NMDA receptors are located post synaptic to adrenergic terminals in the spinal cord. Adrenergic terminals were labeled using an antibody directed against phenylethanolamine-N-methyl transferase (PNMT) and the NMDA receptor was identified using an antibody directed against the R1 subunit of the receptor (NMDAR1). NMDAR1 was found primarily in large and small dendrites and a few perikarya. The presence of NMDAR1 in the dendritic targets of PNMT-containing terminals was quantified in spinal cords sectioned either horizontally or coronally. PNMT-labeled terminals formed asymmetric synapses on dendrites containing immunogold labeling for NMDAR1, but NMDAR1 was more often detected in the targets of PNMT terminals when spinal cords were sectioned horizontally (59%) rather than coronally (28%). This difference in prevalence of NMDAR1 in targets of PNMT terminals is likely due to the preferential orientation of SPN dendrites in the horizontal plane, since longer dendritic shafts were visible in horizontal sections. When NMDAR1 was present in the dendritic targets of many adrenergic terminals, it was usually located at sites distal to the adrenergic input. We conclude that NMDA receptor ligands are likely to modulate the activity of dendritic targets of adrenergic terminals in the spinal cord, but are not closely associated with adrenergic synaptic input. PMID- 10677606 TI - C-terminal phosphorylation of the high molecular weight neurofilament subunit correlates with decreased neurofilament axonal transport velocity. AB - We probed the relationship of NF axonal transport of neurofilaments (NFs) to their phosphorylation state by comparing these parameters in two closely-aged groups of young adult mice - 2 and 5 months of age. This particular time interval was selected since prior studies demonstrate that optic axons have already completed axonal caliber expansion and attained adult NF levels by 2 months but, as shown herein, continue to increase NF-H C-terminal phosphorylation. NF axonal transport was monitored by autoradiographic analysis of the distribution of radiolabeled subunits immunoprecipitated from optic axon segments at intervals following intravitreal injection of 35S-methionine. Both the peak and front of radiolabeled NFs translocated faster in 2- vs. 5-month-old mice. This developmental decline in NF transport rate was not due to reduced incorporation of NFs into the cytoskeleton, nor to an overall decline in slow axonal transport. By excluding or minimizing other factors, these findings support previous conclusions that C-terminal NF phosphorylation regulates NF axonal transport. PMID- 10677607 TI - Elevated substance-P-like immunoreactivity levels in spinal dialysates during the formalin test in normal and diabetic rats. AB - Pharmacologic studies implicate the involvement of substance P in spinal nociceptive processing during the formalin test. However, no direct measurement of the temporal changes in substance P levels within the spinal cord of conscious animals has been reported. Further, dissociation between substance P levels and formalin-evoked nocifensive behavior may exist in diabetic rats, as exaggerated hyperalgesic behavior coexists with reduced peripheral nerve substance P levels. The present study was performed to directly measure the appearance of substance-P like immunoreactivity (SP-LI) in spinal CSF of conscious, unrestrained rats using microdialysis techniques following injection of formalin into the hindpaw. The effect of diabetes upon formalin-evoked SP-LI levels in spinal CSF dialysates was also determined. In control rats, SP-LI increased in spinal dialysates following formalin injection and levels were maximal 20-30 min after injection, rising to 325% of basal values (p<0.02). Diabetic rats exhibited reduced (p<0.05) SP-LI in their spinal roots, while basal levels in spinal CSF were not different from controls. Formalin-evoked nocifensive behavior was increased in diabetic rats but SP-LI levels in spinal CSF dialysates after paw formalin injection were significantly (p<0.05) attenuated, reaching a maximum of only 161% of basal levels. This was accompanied by attenuated swelling at the formalin injection site and increased thermal response latencies. While increased SP-LI in spinal CSF coincides with phase 2 behavior in the formalin test and may contribute to spinal nociceptive processing during this period, exaggerated spinal substance P release is unlikely to underlie the increased nocifensive behavior seen in diabetic rats. PMID- 10677608 TI - Reduced glutathione oxidation ratio and 8 ohdG accumulation by mild ischemic pretreatment. AB - A critical role of oxidative stress has been implicated in ischemic brain damage. Mild ischemic pretreatment and/or synthesis of heat shock proteins (HSPs) has been suggested to protect against oxidative brain damage. However, experimental support of this suggestion have proven to be difficult partly because sensitive indices to assess oxidative consequences of ischemic brain damage were few. In this study, we have attempted to establish biochemical assay systems to quantitate oxidative brain damage following ischemia. We produced experimental brain ischemia in the Mongolian gerbil (Meriones unguiculatus) and examined the hippocampus for ischemic brain damage. The results obtained from ischemic gerbil hippocampus demonstrated that oxidative brain damage can be quantitated by determining glutathione oxidation ratio together with the accumulation of the oxidative DNA damage product, 8-hydroxy-2'-deoxyguanosine (8 ohdG). Our results also demonstrated a role for mild ischemic pretreatment and synthesis of HSPs against oxidative brain damage. We showed that mild 2-min ischemic pretreatment reduced the degree of both glutathione oxidation ratio and 8 ohdG accumulation in gerbil hippocampus subsequent to 10 min ischemic challenge. We also showed that the accumulation of HSP70 was closely associated with the reduction of oxidative brain damage. To our knowledge, this is the first report to investigate glutathione redox states and oxidative DNA damage levels to evaluate a protective role of mild ischemic pretreatment and HSP synthesis following brain ischemia. Our data validate the previous suggestions and provide new additional data that argue for the protective role of mild ischemic pretreatment and HSP70 synthesis against oxidative brain damage. PMID- 10677609 TI - Localization of hindbrain glucoreceptive sites controlling food intake and blood glucose. AB - Feeding and blood glucose responses to local injection of nanoliter volumes of 5 thio-D-glucose (5TG), a potent antimetabolic glucose analogue, were studied at 142 hindbrain and 61 hypothalamic cannula sites. A site was considered positive if 5TG elicited at least 1.5 g more food intake or a hyperglycemic response at least 25 mg/dl greater than the respective responses elicited by vehicle injection in the same rat. Of 61 hypothalamic cannula sites tested, none were positive for blood glucose and only one was positive for feeding. Increasing the 5TG dose to 48 ug did not produce additional positive results at hypothalamic sites. In contrast, 66 hindbrain sites were positive for feeding and 49 were positive for blood glucose, with 33 of these being positive for both responses. The distribution of positive sites for feeding and hyperglycemia overlapped almost completely. Positive sites were concentrated in two distinct zones: one in the ventrolateral and one in the dorsomedial medulla. In both locations, the glucoreceptive areas extended approximately from the level of the area postrema (AP) to the pontomedullary junction. Glucoreceptive zones were co-distributed with epinephrine cell groups C1-C3, suggesting that epinephrine neurons may be important components of the neural circuitry for glucoregulation. Localization of glucoreceptive sites will facilitate positive identification of glucoreceptor cells and the direct analysis of the neural mechanisms through which they influence food intake and metabolic responses. PMID- 10677610 TI - Effects of a novel neurotensin peptide analog given extracranially on CNS behaviors mediated by apomorphine and haloperidol. AB - Neurotensin (NT) is a neuropeptide neurotransmitter in the central nervous system. It has been implicated in the therapeutic and in the adverse effects of neuroleptics. Activity of NT in brain can only be shown by direct injection of the peptide into that organ. However, we have developed a novel analog of NT(8 13), NT69L, which is active upon intraperitoneal (i.p.) injection. Like atypical neuroleptics, NT69L blocked the climbing behavior in rats, but not the licking and sniffing behaviors of a high dose (600 microgram/kg) of the non-selective dopamine agonist apomorphine. Its blockade of climbing was very potent with an ED(50) (effective dose at 50% of maximum) of 16 microgram/kg. Both apomorphine and NT69L caused a long-lasting hypothermia, which was greater with the peptide but not synergistic in combination with apomorphine. The ED(50) of NT69L for hypothermia was 390 microgram/kg. NT69L (up to 5 mg/kg i.p.) did not produce catalepsy. However, when given before haloperidol, NT69L, but not clozapine, completely prevented catalepsy. When given after haloperidol, NT69L, but not clozapine, reversed haloperidol's cataleptic effects with an ED(50) of 260 microg/kg. There was no significant difference between the ED(50)s for hypothermia and anticataleptic effects of NT69L. However, the ED(50) for blocking the effects of apomorphine was significantly lower than the other two. These data suggest that NT69L may have neuroleptic properties in humans and may be useful in the treatment of extrapyramidal side effects caused by typical neuroleptics such as haloperidol. PMID- 10677611 TI - Colocalization of arginine-vasotocin and chicken luteinizing hormone-releasing hormone-I (cLHRH-I) in the preoptic-hypothalamic region of the chicken. AB - To characterize a possible relationship between chicken luteinizing hormone releasing hormone-I (cLHRH-I) and arginine-vasotocin (AVT) we performed immunocytochemical double-stainings throughout the preoptic-hypothalamic region of the chicken brain. This study clearly reveals a partial colocalization between both neuropeptides. Single-labeled neurons, containing either cLHRH-I or AVT are found intermingled with double stained cells, immunoreactive (ir) for both peptides. A significant number of double-labeled perikarya is found in the preoptic area, more specifically in the ventral and external portion of the supraoptic nucleus (SOv and SOe) and in the medial preoptic nucleus (MPOv). At the level of the anterior hypothalamus, double-labeled cells are predominantly observed near the third ventricle in the nucleus paraventricularis magnocellularis (PVN) and the nucleus periventricularis hypothalami (PHN). Next to this colocalization, a number of cLHRH-I-ir cell bodies are found in close apposition to AVT-ir fiber profiles in the very same areas. Taken together, these data are the first to provide morphological evidence indicating that the AVT system might be involved in the regulation of cLHRH-I release and thus of reproductive functions in birds. PMID- 10677612 TI - Selective responsiveness of medial prefrontal cortex neurons to the meaningful stimulus with a low probability of occurrence in rats. AB - Multi-unit neuronal activity was recorded in the medial prefrontal cortex (mPFC) of 13 chronically prepared male rats while they performed a two-tone discrimination task. Tones at 1000 and 2000 Hz were sequentially presented at intervals of 3-6 s. The duration of each tone was 0.8 s. Rats were trained to press a bar within 1.2 s after the cessation of the 1000 Hz tone (target), and not to press the bar when the other tone (non-target) was presented. Intracranial electrical stimulation (ICS) of the medial forebrain bundle was given as a reward immediately after the rats had correctly responded to the target tone. Probability of the target occurrence was either 30% or 70% in different sessions. When the target tone was presented on only 30% of the trials, the mPFC neurons in the majority of rats tested (10/13) exhibited phasic excitation about 100 ms after the onset of the target tone. However, when the target tone occurred on 70% of the trials, mPFC neurons in most of rats (11/13) did not show excitatory responses, and in some of them (5/13) were inhibited. No mPFC neurons exhibited significant responses to the non-target tone, regardless of its probability. These results suggest that the mPFC neurons selectively respond to meaningful events with a low probability of occurrence. PMID- 10677613 TI - Cloning, sequencing, chromosomal location, and function of cDNAs encoding an opioid growth factor receptor (OGFr) in humans. AB - The native opioid growth factor (OGF), [Met(5)]-enkephalin, is a tonic inhibitory peptide that modulates cell proliferation and tissue organization during development, cancer, cellular renewal, wound healing, and angiogenesis. OGF action is mediated by a receptor mechanism. We have cloned and sequenced cDNAs encoding multiple spliced forms of a human OGF receptor. The open reading frame in the longest cDNA was found to encode a protein of 697 amino acids, and 8 imperfect repeats of 20 amino acids each were a prominent feature. Altogether, five alternatively spliced forms were observed. The cDNA hybridized to mRNA from a variety of normal and neoplastic cells and tissues. Functional studies using antisense oligonucleotides to OGFr demonstrated an enhancement in cell growth. Fluorescent in situ hybridization (FISH) experiments showed the chromosomal location to be 20q13.3. This OGF receptor has no homology to classical opioid receptors. These results provide molecular validity for the interaction of OGF and OGF receptor in the regulation of growth processes in humans. PMID- 10677614 TI - The distribution of neuronal nitric oxide synthase in the nucleus tractus solitarii of the squirrel monkey. AB - The distribution of neuronal nitric oxide synthase (nNOS) containing neurons and fibers in subnuclei of the nucleus tractus solitarii (NTS) in the squirrel monkey, Saimuri sciureus, was investigated by nNOS immunohistochemistry and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. Generally, the staining pattern of nNOS and NADPH-diaphorase in the NTS was similar. A high density of neurons and fibers exhibiting both nNOS immunoreactivity and NADPH-diaphorase reactivity was present in the central, medial, intermediate, and dorsolateral subnuclei of the NTS. A moderate density of neurons and fibers that stained for both nNOS and NADPH-diaphorase was noted in the interstitial and ventromedial subnuclei. The gelatinosus and commissural subnuclei contained a low density of neurons and fibers exhibiting nNOS immunoreactivity and NADPH-diaphorase staining. The dorsal motor nucleus of vagus contained a high density of nNOS immunopositive and NADPH-diaphorase containing neurons and fibers at the rostral level, but contained a moderate density of positive fibers and very few positive neurons at the intermediate, subpostremal and commissural NTS levels. Incongruence was noted, however, between nNOS immunostaining and NADPH-diaphorase staining in blood vessels in the brainstem. Capillaries and small vessels exhibited strong staining for NADPH-diaphorase but no nNOS immunoreactivity. In summary, this work substantiates the presence of nNOS in subnuclei of the monkey NTS and is consistent with a role for NO(.) in neurotransmission in primate NTS. PMID- 10677615 TI - Early and sequential recruitment of apoptotic effectors after focal permanent ischemia in mice. AB - In experimental models of cerebral ischemia, cells within the damaged territory die by necrosis and by apoptosis that contributes to the expansion of the insult. Apoptotic machinery mobilizes intracellular processes such as induction of Bcl-2 family members, activation of the proteolytic cascade including the caspases, and cleavage of caspase substrates, such as poly(ADP-ribose) polymerase or PARP. Mitochondria play a pivotal role in controlling apoptosis by releasing cytochrome c and modulating redox state, both under the regulation of manganese superoxide dismutase (Mn SOD) via superoxide anion detoxification. The implication and the kinetics of such events in apoptosis induced after focal permanent ischemia in mice remains to be studied. In a paradigm of ischemic insult induced by occlusion of the middle cerebral artery (MCAO) in mice, we showed by immunohistochemistry a constitutive expression of caspase-3 that is enhanced after MCAO in neurons localized within the infarcted zone. As a function of time intervals after MCAO, the cytochrome c amount increased in the cytosolic fraction of ischemic cortical extracts. The kinetics of the release was in concordance with the expression of caspase-3 and the subsequent cleavage of PARP appearing before the internucleosomal fragmentation of DNA, the ultimate step of apoptosis. When the apoptotic markers progressively appeared, no changes of Mn SOD activity or Mn SOD expression were detected after MCAO. We can therefore speculate that the recruitment of Mn SOD did not participate per se in the release of cytochrome c elicited after permanent focal ischemia. PMID- 10677616 TI - Mechanical activation of dorsal root ganglion cells in vitro: comparison with capsaicin and modulation by kappa-opioids. AB - The aim of this study was to characterize plasma membrane pathways involved in the intracellular calcium ([Ca(2+)](i)) response of small DRG neurons to mechanical stimulation and the modulation of these pathways by kappa-opioids. [Ca(2+)](i) responses were measured by fluorescence video microscopy of Fura-2 labeled lumbosacral DRG neurons obtained from adult rats in short-term primary culture. Transient focal mechanical stimulation of the soma, or brief superfusion with 300 nM capsaicin, resulted to [Ca(2+)](i) increases which were abolished in Ca(2+)-free solution, but unaffected by lanthanum (25 microM) or tetrodotoxin (10(-6) M). 156 out of 465 neurons tested (34%) showed mechanosensitivity while 55 out of 118 neurons (47%) were capsaicin-sensitive. Ninty percent of capsaicin sensitive neurons were mechanosensitive. Gadolinium (Gd(3+); 250 microM) and amiloride (100 microM) abolished the [Ca(2+)](i) transient in response to mechanical stimulation, but had no effect on capsaicin-induced [Ca(2+)](i) transients. The kappa-opioid agonists U50,488 and fedotozine showed a dose dependent inhibition of mechanically stimulated [Ca(2+)](i) transients but had little effect on capsaicin-induced [Ca(2+)](i) transients. The inhibitory effect of U50,488 was abolished by the kappa-opioid antagonist nor-Binaltorphimine dihydrochloride (nor-BNI; 100 nM), and by high concentrations of naloxone (30-100 nM), but not by low concentrations of naloxone (3 nM). We conclude that mechanically induced [Ca(2+)](i) transients in small diameter DRG somas are mediated by influx of Ca(2+) through a Gd(3+)- and amiloride-sensitive plasma membrane pathway that is co-expressed with capsaicin-sensitive channels. Mechanical-, but not capsaicin-mediated, Ca(2+) transients are sensitive to kappa opioid agonists. PMID- 10677617 TI - Methylmalonic and propionic acids increase the in vitro incorporation of 32P into cytoskeletal proteins from cerebral cortex of young rats through NMDA glutamate receptors. AB - In this study we investigated the effects of methylmalonic acid (MMA) and propionic acid (PA) on the phosphorylation of cytoskeletal proteins of cerebral cortex of rats. Slices of tissue were incubated with 32P-orthophosphate in the presence or absence of glutamate, MMA, PA and ionotropic or metabotropic glutamate receptor agonists. The cytoskeletal fraction was isolated and the radioactivity incorporated into the cytoskeletal proteins was measured. Results demonstrated that the acids, glutamate and NMDA increased the phosphorylation of the proteins studied. However, this effect was not observed for non-NMDA ionotropic agonists or metabotropic agonists. Experiments using glutamate receptor antagonists confirmed that MMA and PA at the same concentrations as found in tissues from propionic or methylmalonic acidemic children increase the phosphorylation of cytoskeletal proteins, possibly via NMDA glutamate receptors. Therefore, it is feasible that these findings may be related to the neurological dysfunction characteristic of these disorders. PMID- 10677618 TI - Effects of gestational hypoxia on mRNA levels of Glut3 and Glut4 transporters, hypoxia inducible factor-1 and thyroid hormone receptors in developing rat brain. AB - Alterations of brain development result from noxious intrauterine signals, as oxygen deprivation, which decrease glucose energetic yield. To verify the hypothesis that a defect of brain energetic adaptation is responsible for these alterations, we have studied the effects of gestational hypoxia (10% oxygen during the last 2 weeks of fetal life) on cerebral ontogenesis of glucose transporters which control the limiting step of glucose utilization by neurons. This study is realised in rats by quantification of whole brain Glut3 and Glut4 mRNA in 14- and 19-day-old embryos (E14, E19), newborn (P0) and 7 postnatal-day old rats (P7) by using reverse transcription-polymerase chain reaction (RT-PCR) method. We have associated our study with the analysis of a transcriptional factor, the hypoxia inducible factor-1alpha (HIF-1alpha), known to control the expression of glucose transporter, and with a family of transcriptional factors, the thyroid hormone receptors (TR), regulating specific genes involved in brain development. The data show (1) for the first time the Glut4 and HIF-1alpha gene expression in fetal rat brain which are detected as soon as E14, (2) that gestational hypoxia induces an increase of mRNA transcript levels of Glut3, Glut4, TRalpha2, TRbeta1 and HIF-1alpha genes mainly or exclusively at E14, and (3) that the absence of response of Glut3 and HIF-1alpha at E19 in hypoxic vs. normoxic group could indicate an insufficient energetic adaptation at this period of development which could lead to the neural alterations observed postnatally. PMID- 10677619 TI - LP-BM5 infection impairs spatial working memory in C57BL/6 mice in the Morris water maze. AB - Previous studies show that the LP-BM5 murine leukemia virus causes an acquired immunodeficiency syndrome in C57BL/6 mice (MAIDS) and impairs learning and memory without gross motor impairment. To assess spatial working memory impairment after LP-BM5 infection and the time course of this impairment, we tested mice in a modified working-memory version of the Morris water maze. Twenty mice were inoculated with LP-BM5; controls received medium (Minimum Essential Medium). In the test procedure, animals had two 1-min training sessions to learn the position of a randomly placed hidden platform. Thirty seconds after the second training session, animals were placed in the maze without the platform, and time and pathlength spent in each quadrant of the maze were measured. For 9 weeks after LP BM5 infection, both groups showed preference for the target quadrant compared to the opposite quadrant. At 10 and 11 weeks after infection, the LP-BM5 virus infected mice lost this target quadrant preference. We conclude that LP-BM5 infection impaired spatial working memory in a modified working-memory version of the Morris water maze test in C57BL/6 mice at 10 and 11 weeks after virus infection. PMID- 10677620 TI - Sensory deafferentation and olfactory bulb morphology in the zebrafish and related species. AB - The zebrafish, Danio rerio, has become an important model species for examining olfactory system structure and function, yet little is known about developmental changes in olfactory bulb morphology from embryo to adult. The present study examined both normal growth and the effects of deafferentation on the bulb from hatching to adulthood. In young animals, the bulb is small relative to body size and has a higher percentage of its volume occupied by incoming olfactory nerve fibers. Young animals are also more affected by sensory deafferentation. Olfactory rosette removal resulted in more than 50% reductions in laminar volumes, indicating that sensory input is important during periods of rapid development. In addition, three closely related species were examined to compare how differing bulb morphology might influence the effects of bulb manipulation. The cherry barb, Barbus (=Puntius) titteya, and giant danio, Danio aequipinnatus, have larger bulbs and laminar volumes relative to body size than the zebrafish or scissortail rasbora, Rasbora trilineata. Both are also more affected by deafferentation, with at least a 35% reduction in laminar sizes in many of the bulb layers. The studies are discussed in terms of the importance of the olfactory system to each species and are also compared to the effects of sensory manipulations in other animals. PMID- 10677621 TI - Cells containing immunoreactive estrogen receptor-alpha in the human basal forebrain. AB - The distribution of estrogen receptor protein-alpha (ER-alpha)-containing cells in the human hypothalamus and adjacent regions was studied using a monoclonal antibody (H222) raised against ER-alpha derived from MCF-7 human breast cancer cells. Reaction product was found in restricted populations of neurons and astrocyte-like cells. Neurons immunoreactive for ER-alpha were diffusely distributed within the basal forebrain and preoptic area, infundibular region, central hypothalamus, basal ganglia and amygdala. Immunoreactive astrocyte-like cells were noted within specific brain regions, including the lamina terminalis and subependymal peri-third-ventricular region. These data are consistent with the location of estrogen receptors in the basal forebrain of other species and the known effects of estrogens on the cellular functions of both neurons and supporting elements within the human hypothalamus and basal forebrain. PMID- 10677622 TI - Organization of intratelencephalic projections to the visual Wulst of the chick. AB - The avian visual Wulst, said to be the equivalent of the striate cortex in mammals, is the telencephalic visual area of the thalamofugal visual pathway. In this study, by means of retrograde labelling with fluorescent tracers injected into the Wulst regions in the left and right hemispheres, we have investigated the organization of the intratelencephalic projections to the visual Wulst in chicks. After injecting Fluorogold (FG), True blue (TB) or rhodamine into the visual Wulst, fluorescent-labelled neurones were found in the ipsilateral neostriatum frontale, pars lateralis (NFl), the ipsilateral neostriatum intermedium (NI) and the ipsilateral dorso-lateral neostriatum. Labelled neurones were also found in both the ipsilateral and contralateral archistriata. In addition, some neurones in the archistriatum were double-labelled, which indicates that these archistriatal neurones have axon collaterals projecting to the visual Wulst on both sides of the forebrain. Through these intratelencephalic afferents to the visual Wulst, visual information transmitted in the thalamofugal pathway may be modulated by other telencephalic areas. The possible roles of these connections in regulating behaviour are discussed. PMID- 10677623 TI - Conformation of paired helical filaments blocks dephosphorylation of epitopes shared with fetal tau except Ser199/202 and Ser202/Thr205. AB - To determine if the high phosphate content of paired helical filaments (PHFs) in Alzheimer's disease (AD) is a result of limited access to filament phosphorylation sites, we studied in vitro dephosphorylation of intact PHFs, PHFs with filamentous structure abolished by formic acid treatment (PHF(FA)) and fetal human tau protein. Samples were treated with alkaline phosphatase for up to 24 h at 37 degrees C and then immunoblotted with eight well characterized tau antibodies, that recognize two phosphorylation-insensitive sites and six phosphorylation-sensitive epitopes at Thr181, Ser199/202, Ser202/Thr205, Thr231, Ser262/356 and Ser396/404. Intact PHFs were effectively dephosphorylated only at the two N-terminal epitopes Ser199/202 and Ser202/Thr205, with little change in electrophoretic mobility. In contrast, PHF(FA) were dephosphorylated at all epitopes, with particular effectiveness at those in the C-terminus and with significant increase in electrophoretic mobility. The fetal tau epitopes were effectively dephosphorylated except at Thr181 and Thr231 with marked increase in mobility. The extent of dephosphorylation of PHF(FA) was equal or more effective than in fetal tau, except for Thr181 that was minimally dephosphorylated in both proteins. The results indicate that intact PHFs, but not PHF(FA) or fetal tau display differential dephosphorylation of the N- and C-terminal epitopes. The results confirm that the filamentous conformation may significantly contribute to hyperphosphorylation of PHFs in the C-terminus. The filamentous conformation, however, does not limit access to two N-terminal epitopes Ser199/202 and Ser202/Thr205. The access to these sites in AD may be limited by other factors, e.g., inhibition of phosphatase binding. PMID- 10677624 TI - The role of nigral and thalamic output pathways in the expression of oral stereotypies induced by amphetamine injections into the striatum. AB - Microinjections of amphetamine into the ventrolateral striatum (VLS) elicit a striking behavioral syndrome characterized by compulsive oral and forelimb motor stereotypies. The neural pathways that mediate these behavioral responses downstream from the striatum have not yet been identified. In a series of experiments, we investigated the involvement of the substantia nigra pars reticulata (SNr) and the ventromedial nucleus of the thalamus (VMT) in the mediation of this behavioral syndrome. We demonstrated that lidocaine-induced reversible inactivation of the SNr reduced amphetamine-induced stereotyped biting and gnawing behaviors, suggesting that the nigral output pathway plays a significant role in the expression of these behavioral responses. In turn, injections of lidocaine into the VMT only transiently reduced amphetamine stimulated biting and increased stereotyped gnawing and paw nibbling, suggesting that the expression of oral stereotypies induced by amphetamine injections into the VLS is not dependent on thalamocortical feedback. PMID- 10677625 TI - Fetal alcohol exposure alters serotonin transporter sites in rat brain. AB - This study examined the effects of fetal alcohol exposure (FAE) on serotonin transporter (5-HTT) binding sites in the brains of developing male and female rat offspring using the technique of quantitative autoradiography. Time-pregnant dams were fed liquid ethanol diet, isocaloric diet without ethanol or normal rat chow. Male and female offspring were sacrificed at 21, 40 and 60 days of age, brains removed and sectioned for analysis of 5-HTT sites. FAE led to distinct effects on 5-HTT sites depending on the age and gender of the offspring. FAE increased 5-HTT binding sites in cortical layers 5, 6, hippocampal layers CA(2,3), lateral nucleus of the amygdala and in the dorsal raphe nucleus. FAE decreased 5-HTT binding sites in the medial nucleus of amygdala, dorsomedial and ventromedial nuclei of the hypothalamus. FAE decreased 5-HTT binding sites temporarily in the ventromedial nucleus of the hypothalamus in the 21-day-old female; this effect was found to disappear by day 40. In contrast, FAE increased 5-HTT sites in the lateral nucleus of the amygdala in the adult animal, suggesting that ethanol exposure in utero may alter serotonin neurotransmission in discrete brain regions permanently. PMID- 10677626 TI - Microinfusion of protein kinase inhibitor H7 into the cerebellum impairs the acquisition but not the retention of classical eyeblink conditioning in rabbits. AB - Rabbits were infused with H7, a general protein kinase inhibitor, into the region of the cerebellar interpositus nucleus during classical eyeblink conditioning. Acquisition of the conditioned eyeblink response was delayed by the H7 infusion, but the protein kinase inhibitor had no effect on performance of the learned response when infused after asymptotic learning had been reached. These data indicate that protein kinases in the cerebellum are involved in plasticity processes that underlie the learning of this simple conditioned behavior. PMID- 10677627 TI - Morphologic and electrophysiologic maturation in developing dentate gyrus granule cells. AB - Dentate gyrus granule cells from immature (7-28 days) Sprague-Dawley rats were examined with whole cell patch clamp recordings and biocytin filling in in vitro hippocampal slice preparations. Although recordings were confined to the middle third of the suprapyramidal limb of the dentate, the granule cells exhibited marked variability in their physiologic properties: input resistance (IR) ranged from 250 MOmega to 3 GOmega, and resting membrane potential (RMP) from -82 to -41 mV. Both IR and RMP were inversely correlated with dendritic length, a morphometric indicator of cell maturity. Thus the highest IR cells were the youngest, and maturation was characterized by a progressive decrease in IR, hyperpolarization of RMP, and elongation of the dendritic arbor. When cells were grouped by IR, significant intergroup differences were found in RMP, dendritic length, and number of dendritic terminal branches. Although cells of all IR categories were examined throughout the age spectrum under study, none of the inter-IR group differences was age-dependent. These data suggest that IR provides a reasonable estimate of granule cell maturity and that maturation entails predictable changes in cell properties and morphology. These aspects of maturation correlate with each other, are independent of animal age, and most likely proceed according to a program related to cell birth. PMID- 10677628 TI - Aging regulates 5-HT(1B) receptors and serotonin reuptake sites in the SCN. AB - Middle age is associated with changes in circadian rhythms (e.g., alterations in the timing of the circadian wheel running rhythm) which resemble changes induced by selective destruction of the serotonergic input to the suprachiasmatic nucleus (SCN), the principal mammalian circadian pacemaker. We hypothesized that serotonergic neurotransmission in the SCN is decreased in middle-aged hamsters, as compared to young adults. This hypothesis was tested indirectly by investigating the effect of aging on two markers of serotonin neurotransmission, 5-HT(1B) receptors and serotonin reuptake sites, which are regulated by serotonin. Previous studies have shown that experimentally induced decreases in serotonergic neurotransmission increase 5-HT(1B) receptors but decrease serotonin reuptake sites. Quantitative autoradiography was conducted using [125I]iodocyanopindolol ([125I]ICYP) and [3H]paroxetine, selective radioligands for the 5-HT(1B) receptors and the serotonin reuptake sites, respectively. Consistent with the hypothesis, specific ([125I]ICYP binding was significantly elevated in the SCN of middle-aged hamsters, as compared to young hamsters. The results also showed that serotonin reuptake sites in the SCN were significantly increased in both middle-aged and old hamsters, as compared to young controls. This result could not have been caused by decreased serotonin release. Alternatively, increased serotonin reuptake, which would reduce serotonin levels in the synaptic cleft, may cause or contribute to the increase in 5-HT(1B) receptor binding in the SCN in middle aged animals. These results show that the SCN exhibits changes in serotonergic function during middle age, which has been characterized by changes in the expression of circadian rhythms. Because these changes occur during middle age, they probably reflect the aging process, rather than senescence or disease. PMID- 10677629 TI - Is intracellular brain pH a dependent factor in NOS inhibition during focal cerebral ischemia? AB - The interaction between nitric oxide (NO.) and focal cerebral ischemia is multifaceted. Experiments have shown that inhibition of nitric oxide synthase (NOS) either ameliorates or exacerbates focal cerebral ischemia. Recent in vitro experiments have shown that NOS activity is pH-dependent. Previous work from this laboratory has demonstrated that N(G)-nitro-L-arginine-methyl-ester (L-NAME) mitigated cerebral ischemia independent from regional cerebral blood flow (rCBF) changes during moderate focal cerebral ischemia. This study examined the effects of L-NAME inhibition on brain pH(i), rCBF, and NADH redox state during 3 h of severe focal cerebral ischemia. Fifteen fasted rabbits under 1.5% halothane were equally divided into three groups: ischemic controls and two drug groups receiving either 1.0 or 10 mg/kg L-NAME intravenously 30 min prior to ischemia. In the ischemic controls, brain pH(i) declined from 6.95+/-0.04 to 6.60+/-0.05, rCBF declined from 48+/-7 to 10+/-3 ml/100 g/min, and NADH fluorescence increased by 149+/-15% 3 h after onset of ischemia (p<0.01 for all three parameters). L NAME at either dose did not significantly alter these values. Infarct volume was not significantly different between both the L-NAME treated groups and the ischemic control group. This data suggests that during severe focal cerebral ischemia, NO. mechanisms of injury have a less important punitive role. One possible explanation is that the severity of acidosis secondary to anaerobic metabolism during severe focal cerebral ischemia attenuates NOS activity. PMID- 10677630 TI - Enhanced analgesic potency and reduced tolerance of morphine in 129/SvEv mice: evidence for a deficiency in GM1 ganglioside-regulated excitatory opioid receptor functions. AB - 10-fold higher doses in SW mice. Furthermore, cotreatment of 129/SvEv mice with morphine plus a low dose of naltrexone (ca. 0.1 microgram/kg) that markedly enhances and prolongs morphine's antinociceptive effects in SW mice did not enhance, and often attenuated6 h. The marked GM1-induced attenuation of morphine's antinociceptive effects in 129/SvEv mice may be due to conversion of some of the opioid receptors in these mice from an inhibitory Gi/Go-coupled to an excitatory Gs-coupled mode. Exogenous GM1 supplementation can, therefore, reverse the anomalous lack of morphine tolerance displayed by this mouse strain in comparison to SW and other mice. The present study may provide insights into factors that regulate the marked variability in nociceptive sensitivity and opioid tolerance/dependence liability among individual humans. PMID- 10677631 TI - Flupirtine ameliorates ischaemic-like death of rat retinal ganglion cells by preventing calcium influx. AB - The effect of flupirtine on the loss of retinal ganglion cells following transient elevation of intraocular pressure (experimental ischaemia) or NMDA induced excitotoxicity was studied. Ischaemia (60 min) or intravitreal injection of NMDA (20 nmol) caused a decrease in Thy-1 mRNA and Thy-1 immunoreactivity which are associated with ganglion cells. Administration of flupirtine counteracted these changes. Moreover, flupirtine dose-dependently inhibited NMDA induced 45Ca(2+) influx into cultured cortical neurones and retinal pieces in vitro with maximal inhibition being observed at 200 microM. A similar concentration of flupirtine failed to inhibit kainate-stimulated calcium influx into cultured cortical neurones. In addition, flupirtine had no significant effect on [3H]nitrendipine or [3H]diltiazem binding to cortical membranes. The present studies are consistent with previous findings which suggested flupirtine to act as a NMDA antagonist by a mechanism that still remains to be clarified. PMID- 10677632 TI - Discharge of group IV phrenic afferent fibers increases during diaphragmatic fatigue. AB - The discharge of single unit group III (n=7) and group IV (n=8) phrenic afferent fibers was recorded during rhythmic diaphragmatic contractions before and after the onset of fatigue. Compared to pre-fatigue impulse activity, group IV, but not group III, phrenic afferent fibers discharged more (p<0.05) during rhythmic diaphragmatic contractions when the diaphragm was fatigued. This increase in group IV fiber discharge during diaphragmatic fatigue provides electrophysiological evidence consistent with the notion that group IV phrenic afferent fibers comprise the afferent arm of a fatigue-induced inhibitory reflex originating in the diaphragm. PMID- 10677633 TI - Clonidine diminishes c-jun gene expression in the cardiovascular sensitive areas of the rat brainstem. AB - The present study investigated the effect of clonidine on the basal and inducible c-jun and c-fos mRNA expression in the nucleus tractus solitarius (middle, mNTS, and rostral, rNTS) and the rostral ventrolateral medulla (caudal, cRVLM, and rostral, rRVLM). Conscious rats received saline, clonidine (30 microg/kg, i.v.), saline plus sodium nitroprusside (NP), or clonidine plus NP. Under basal conditions (saline-infused rats), c-jun mRNA was expressed in the mNTS and rRVLM but not in the rNTS or cRVLM whereas c-fos mRNA was not detectable. Clonidine attenuated the increases in c-fos in the mNTS and cRVLM and c-jun gene expression in the mNTS and rRVLM caused by NP-evoked hypotension and also reduced the basal expression of c-jun mRNA in the mNTS and rRVLM. These findings establish a causal link between clonidine inhibition of c-fos expression in brainstem and its hypotensive action, and provide the first evidence that clonidine attenuates the expression of the closely linked c-jun gene in neurons implicated in centrally mediated hypotension. PMID- 10677634 TI - Neostigmine influences the L-dopa-induced extracellular dopamine levels in the striatum. AB - Using in vivo microdialysis in freely moving rats, we show that the addition to the dialysis perfusion fluid of the acetylcholinesterase inhibitor neostigmine influences the decarboxylation of levodopa (L-dopa). Continuous perfusion of neostigmine (10, 50 and 100 nM) in striatum attenuated the L-dopa-induced dopamine release in a dose-dependent manner. This effect suggests that changes in magnitude of drug responses may occur when an acetylcholinesterase inhibitor is included in the perfusion solution. Results obtained under these circumstances should be carefully interpreted concerning the pharmacological effects of other drugs when used concomitantly with neostigmine. PMID- 10677635 TI - The interstitial nuclei of the human anterior hypothalamus: an investigation of sexual variation in volume and cell size, number and density. AB - The four interstitial nuclei of the anterior hypothalamus (INAH) have been considered as candidate human nuclei for homology with the much studied sexually dimorphic nucleus of the preoptic area of the rat. Assessment of the INAH for sexual dimorphism has produced discrepant results. This study reports the first systematic examination of all four INAH in the human for sexual variation in volume, neuronal number and neuronal size. Serial Nissl-stained coronal sections through the medial preoptic area and anterior hypothalamus were examined from 18 males and 20 females who died between the ages of 17 and 65 without evidence of hypothalamic pathology or infection with the human immunodeficiency virus. A computer-assisted image-analysis system and commercial stereology software package were employed to assess total volume, neuronal number and mean neuronal size for each INAH. INAH3 occupied a significantly greater volume and contained significantly more neurons in males than in females. No sex differences in volume were detected for any of the other INAH. No sexual variation in neuronal size or packing density was observed in any nucleus. The present data corroborate two previous reports of sexual dimorphism of INAH3 but provide no support for previous reports of sexual variation in other INAH. PMID- 10677636 TI - U-69,593 microinjection in the infralimbic cortex reduces anxiety and enhances spontaneous alternation memory in mice. AB - The present report investigated the contributions of the ventromedial prefrontal cortex to the control of spontaneous alternation/working memory and anxiety related behaviour. In Experiment 1, we examined the effects of microinjections of the selective kappa(1) receptor agonist, U-69,593, in the infralimbic cortex (IL) of CD-1 mice on several ethologically-derived anxiety indices in the elevated plus-maze (EPM) and defensive/withdrawal (D/W) anxiety in the open field, as well as on memory in the EPM transfer-latency (T-L) test and implicit spontaneous alternation memory (SAP) in the Y-maze. In week 1, pretreatment with one injection of vehicle, 1, 10 or 25 nmol/1.0 microliter U-69,593 in the IL dose dependently prolonged T-L and produced a dose-dependent anxiolytic behavioural profile in the first EPM trial. Following a 24-h delay, the same mice were given a drug-free second trial in the EPM tests of T-L memory and anxiety. Whereas T-L memory was not disturbed, small but detectable carry-over effects were observed in trial-2 EPM behaviour relative to vehicle-treated animals. In week 2, the same groups of mice were again pretreated with one injection of the same doses of U 69,593 in the IL and given a D/W test in an open field, followed immediately by an 8-min SAP trial in the Y-maze. The smallest U-69,593 dose was anxiolytic in the D/W test, and SAP/working memory was dose-dependently enhanced in the Y-maze. In Experiment 2, we evaluated whether 0.5 microliter volume microinjections would produce comparable behavioural and carry-over effects in the IL of three new groups of CD-1 mice, in the event that the 1.0 microl volume injections used in Experiment 1 diffused beyond the IL and therefore may have confounded some effects. Experiment 2 procedures were carried out in the same manner as in Experiment 1, except the animals were tested in reverse order. Thus in week 1, SAP memory was tested in the Y-maze followed by D/W anxiety in the open field for half of the animals in each group, and the other half was tested in reverse order. In week 2, T/L memory and anxiety were tested in the EPM in 2 trials as described in Experiment 1. Pretreatment with one injection of vehicle, 10 or 25 nmol/0.5 microliter U-69,593 in the IL reduced D/W anxiety and enhanced SAP memory regardless of testing order in week 1. In week 2, the same groups of mice were again pretreated with one injection of the same doses of U-69,593 in 0.5 microliter volumes in the IL and tested in the EPM. In a similar fashion to Experiment 1, U-69,593 dose-dependently prolonged T/L and produced an anxiolytic behavioural profile in the first EPM trial. Following a 24-h delay, T/L recall memory was again not significantly influenced, but a robust anxiolytic behavioural profile was observed in the second drug-free anxiety trial in the EPM relative to vehicle-treated animals. Results are discussed relative to a) injection volumes and testing order, b) the possible influence kappa receptors may exert on neurochemical responsivity to anxiety-provoking environments in the IL area of the mPFC, c) the possibility that kappa-mediated anxiolysis from the IL in CD-1 mice results from interactions with neurochemical systems involved in the blunting of incoming anxiety-provoking information, d) evidence that SAP memory may be an implicit subtype of working memory, and e) the possibility that IL implicit working memory processes may modulate the induction and expression of anxiety-related behaviour. PMID- 10677637 TI - Comparison of cytosine arabinoside delivery to rat brain by intravenous, intrathecal, intraventricular and intraparenchymal routes of administration. AB - We evaluated the delivery of 14C-cytosine arabinoside (AraC) to rat brain by: 1) intravenous (IV) bolus, by 2) intrathecal (IT) and 3) intraventricular (IVT) infusion, and by 4) convection-enhanced delivery (CED) into the caudate nucleus. Plasma and brain AraC metabolites were measured with HPLC, and distribution and concentration of 14C-AraC in brain sections were measured by quantitative autoradiography. After IV administration, the alpha and beta plasma half-lives were 1.9 and 46.5 min, respectively. The blood-to-brain transfer constant of AraC was 2.5+/-1.4 microliter g(-1) min(-1), compatible with high water solubility. After IT and IVT administration, tissue levels were high at the brain and ventricular surfaces, but declined exponentially into brain. After CED, maximum brain levels were up to 10,000 times higher than the IV group, and the distribution pattern was one of high 14C-AraC concentration in the convective component, with exponentially declining concentrations outside this region. The rate loss constant from brain was 0.002+/-0.0004 min(-1), suggesting that AraC was accumulating in brain cells. AraC was metabolized into uracil arabinoside within the brain. 14C-AraC was infused into 1 dog and distributed widely in the ipsilateral hemisphere. These studies suggest that delivery of AraC to brain parenchyma by the IV, IT or IVT routes will be subtherapeutic. Delivery by CED can achieve, and maintain, therapeutic levels of AraC in the brain, and should be further evaluated as a potential method of drug delivery. PMID- 10677638 TI - Brain activation during human finger extension and flexion movements. AB - Corticospinal projections to the motor neuron pool of upper-limb extensor muscles have been reported to differ from those of the flexor muscles in humans and other primates. The influence of this difference on the central nervous system control for extension and flexion movements is unknown. Cortical activation during thumb extension and flexion movements of eight human volunteers was measured using functional magnetic resonance imaging (fMRI), which detects signal changes caused by an alteration in the local blood oxygenation level. Although the relative activity of the extensor and flexor muscles of the thumb was similar, the brain volume activated during extension was substantially larger than that during flexion. These fMRI results were confirmed by measurements of EEG-derived movement-related cortical potential. Higher brain activity during thumb extension movement may be a result of differential corticospinal, and possibly other pathway projections to the motoneuron pools of extensor and flexor muscles of upper the extremities. PMID- 10677639 TI - Striatal trophic activity is reduced in the aged rat brain. AB - Our previous studies demonstrated that the survival of a mesencephalic graft was reduced in aged animals suggesting an age-related decline in target-derived neurotrophic activity. We tested this hypothesis by examining dopamine (DA) and trophic activities from the striatum of intact or unilateral 6-hydroxydopamine (6 OHDA) lesioned rats of increasing age. Fisher 344 rats were 4, 12, 18, and 23 months old (m.o.) at sacrifice. Half the animals had received unilateral 6-OHDA lesions of the mesostriatal DA pathway 8 weeks earlier. Striatal tissue punches were analyzed for DA, homovanillic acid (HVA), and DA activity (HVA/DA) using HPLC. The remainder of the striatal tissue was homogenized to generate tissue extracts which were added to E14.5 ventral mesencephalic cultures to test trophic activity. In the non-lesioned animals, striatal DA was reduced and striatal DA activity was increased in the 18 and 23 m.o. animals relative to the 4 and 12 m.o. animals. Striatal trophic activity was inversely related to age. In the lesioned animals, striatal DA ipsilateral to 6-OHDA infusion was below detection limits while the contralateral striatum exhibited age-related changes in DA similar to those seen in the non-lesioned animals. In 4 m.o. lesioned rats, striatal trophic activity ipsilateral to 6-OHDA infusion was elevated by 26% relative to the contralateral side. The ipsi/contra-lateral differences in striatal trophic activity were reduced in 12 m.o. animals and absent in the 18 and 23 m.o. groups. These data suggest that advancing age is associated with a reduction in striatal DA as well as trophic activity. Moreover, the aged striatum loses its ability to biochemically and trophically compensate for DA reduction and therefore may represent a more challenging environment for the survival, growth, and function of a fetal graft. PMID- 10677640 TI - The role of the International Organization of Psychophysiology (IOP) in the 20th century and its promising march towards the 21st. AB - In this Presidential Address, the contribution of the International Organization of Psychophysiology (IOP) to the 20th century is summarized. Its official relationship with the United Nations and other international bodies is discussed. Future trends paving the way for the 21st Century as the 'Century of the Brain' are emphasized. The importance of psychophysiology to Humanity is underlined. PMID- 10677641 TI - Brain oscillations in perception and memory. AB - Gamma oscillations, now widely regarded as functionally relevant signals of the brain, illustrate that the concept of event-related oscillations bridges the gap between single neurons and neural assemblies. Taking this concept further, we review experiments showing that oscillatory phenomena such as alpha, theta, or delta responses to events are strongly interwoven with sensory and cognitive functions. This review argues that selectively distributed delta, theta, alpha, and gamma oscillatory systems act as resonant communication networks through large populations of neurons. Thus, oscillatory processes might play a major role in relation with memory and integrative functions. A new 'neurons-brain' doctrine is also proposed to extend the neuron doctrine of Sherrington. PMID- 10677642 TI - Near-infrared spectroscopy: does it function in functional activation studies of the adult brain? AB - Changes in optical properties of biological tissue can be examined by near infrared spectroscopy (NIRS). The relative transparency of tissues including the skull to near-infrared light is the prerequisite to apply the method to brain research. We describe the methodology with respect to its applicability in non invasive functional research of the adult cortex. A summary of studies establishing the 'typical' response in NIRS vascular parameters, i.e. changes in the concentration of oxygenated and deoxygenated haemoglobin, over an activated area is followed by the validation of changes in the cytochrome-oxidase redox state in response to a visual stimulus. Proceeding from these findings a rough mapping of this metabolic response over the motion-sensitive extrastriate visual area is demonstrated. NIRS measures concentration changes in deoxygenated haemoglobin [deoxy-Hb] which are assumed to be the basis of fMRI BOLD contrast (blood oxygenation level-dependent). The method is therefore an excellent tool to validate assumptions on the physiological basis underlying the fMRI signal, due to its high specificity as to the parameters measured. Questions concerning the concept of 'activation'/'deactivation' and that of the linearity of the vascular response are discussed. To challenge the method we finally present results from a complex single-trial motor paradigm study testing the hypothesis, that premotor potentials (contingent negative variation) can be examined by functional techniques relying on the vascular response. Some of the work described here has been published elsewhere. PMID- 10677643 TI - Lonely traits and concomitant physiological processes: the MacArthur social neuroscience studies. AB - Loneliness is a complex set of feelings encompassing reactions to unfulfilled intimate and social needs. Although transient for some individuals, loneliness can be a chronic state for others. Prior research has shown that loneliness is a major risk factor for psychological disturbances and for broad-based morbidity and mortality. We examined differences between lonely and socially embedded individuals that might explain differences in health outcomes. Satisfying social relationships were associated with more positive outlooks on life, more secure attachments and interactions with others, more autonomic activation when confronting acute psychological challenges, and more efficient restorative behaviors. Individuals who were chronically lonely were characterized by elevated mean salivary cortisol levels across the course of a day, suggesting more discharges of corticotropin-releasing hormone and elevated activation of the hypothalamic-pituitary-adrenocorticol axis. An experimental manipulation of loneliness further suggested that the way in which people construe their self in relation to others around them has powerful effects on their self concept and, possibly, on their physiology. PMID- 10677644 TI - Cortical dynamics of memory. AB - Memory networks are formed in the cerebral cortex by associative processes, following Hebbian principles of synaptic modulation. Sensory and motor memory networks are made of elementary representations in cell assemblies of primary sensory and motor cortex (phyletic memory). Higher-order individual memories, e.g. episodic, semantic, conceptual - are represented in hierarchically organized neuronal networks of the cortex of association. Perceptual memories are organized in posterior (post-rolandic) cortex, motor (executive) memories in cortex of the frontal lobe. Memory networks overlap and interact profusely with one another, such that a cellular assembly can be part of many memories or networks. Working memory essentially consists in the temporary activation of a memory network, as needed for the execution of successive acts in a temporal structure of behavior. That activation of the network is maintained by recurrent excitation through reentrant circuits. The recurrent reentry may occur within local circuits as well as between separate cortical areas. In either case. recurrence binds together the associated components of the network and thus of the memory it represents. PMID- 10677645 TI - Visuomotor neurons: ambiguity of the discharge or 'motor' perception? AB - The cortical motor system has been classically considered as the unitary, output stage of the brain processing of sensory information. According to this idea, the motor cortex - the acting brain - receives the result of the perceptual processing (visual, acoustical, tactile, etc.) elaborated by the 'associative cortex'. During the last two decades this perspective has been challenged by a series of anatomical, hodological, and neurophysiological data. This converging evidence delineates a dramatically changed picture. Far from being unitary, the cortical motor system appears to be constituted by a constellation of distinct areas, each of those endowed with specific functional properties and linked by reciprocal connections with distinct sectors of the parietal cortex. Furthermore, several 'motor' neurons in addition to their motor discharge, are also activated by somatosensory and visual stimulation (somatomotor and visuomotor neurons). In the present paper we will discuss the functional properties of those sensorimotor neurons located in the ventral part of the monkey premotor cortex. On the basis of electrophysiological data, we will propose that the apparent parodox stemming from the coexistence within the same neuron of motor and sensory properties can be solved by postulating that the motor system not only executes actions but also internally represents them in terms of 'motor ideas'. These motor ideas may provide the neurobiological basis for space representation, understanding of actions made by others and, possibly, semantic categorization of objects. PMID- 10677646 TI - Hierarchical neuronal modeling of cognitive functions: from synaptic transmission to the Tower of London. AB - Recent progress in the molecular biology of synaptic transmission, in particular of neurotransmitter receptors, offers novel information relevant to 'realistic' modeling of neural processes at the single cell and network level. Sophisticated computer analyses of two-dimensional crystals by high resolution electron microscopy yield images of single neurotransmitter receptor molecules with tentative identifications of ligand binding sites and of conformational transitions. The dynamics of conformational changes can be accounted for by a 'multistate allosteric network' model. Allosteric receptors also possess the structural and functional properties required to serve as coincidence detectors between pre- and post-synaptic signals and, therefore, can be used as building blocks for a chemical Hebb synapse. These properties were introduced into networks of formal neurons capable of producing and detecting temporal sequences. In more elaborate models of pre-frontal cortex functions, allosteric receptors control the selection of transient 'pre-representations' and their stabilization by external or internal reward signals. We apply this scheme to Shallice's Tower of London test, and we show how a hierarchical neuronal architecture can implement executive or planning functions associated with frontal areas. (Academie des sciences/Elsevier, Paris.) PMID- 10677647 TI - Role of neurotrophins in the development and plasticity of the visual system: experiments on dark rearing. AB - An extensive series of studies, beginning with the pioneering experiments of Wiesel and Hubel, have shown that correct visual experience is crucial for the development of the visual system. Several years ago, we put forward the hypothesis that neurotrophic factors of the neurotrophin family (NGF, BDNF, NT-3, NT-4) have a role in mediating the effects of visual experience in the developing visual system. This theory is based on the following experimental results: (a) exogenous supply of neurotrophins during the critical period prevents the effects of monocular deprivation; and (b) transplant of cells releasing NGF allows a normal development of the functional properties of visual cortical neurons in dark-reared rats. PMID- 10677648 TI - Perception and the conditioning reflex: vector encoding. AB - Color perception is dependent on the generation of an excitation vector which, acting on a pool of color detectors (color detector map), produces a corresponding sensation. The generation of the color excitation vector starts at the retinal level, proceeds in the lateral geniculate body, and reaches color detectors at the cortical level. Following processing at the level of declarative memory and semantic maps, results in a verbal categorization of colors. Parallel to the excitation vector pathway, a network computing color differences is operating. The computation of color differences at the retinal level possibly takes place in phasic bipolar cells and progresses in the lateral geniculate body and at the cortical level. Detectors of color differences are assumed to be a basis of respective numerical estimations in humans. Data from frogs, fish, monkeys and humans are compared. PMID- 10677649 TI - Psychophysiology by the end of the 20th century. AB - The first real breakthrough in the research of brain organization and thinking in the 20th century was made in neurophysiological investigations performed in direct contact with different sites of the brain, which became possible in diagnosis and treatment. The second breakthrough is happening at present. It is based on the opportunities provided by the non-invasive technique. The theory of the unique character of the brain system consisting of rigid and flexible elements maintaining thinking was created as well as concepts on the reliability in the system, of the error detector and intrinsic protective mechanisms of the brain. In the clinic these data enabled us to help patients who had lost various functions due to stroke. In confirmation with the above theory it was revealed that the same task could be solved in the brain by systems consisting of different elements due to environmental changes or even direction of attention. Data on the functional properties or every zone of the cortex and subcortex as well as cerebellum are rapidly increasing in number. The first priority lies in neurophysiologically penetrating into the physiological character and micromosaic of the activation sites of PET. The main aim of future brain research lies in the investigation of the fine physiological rearrangements which underlie thinking, i.e. deciphering its brain code. This is going to be the basis for the third, extremely valid breakthrough in the research on brain organization of thinking. PMID- 10677650 TI - Neuronal imprinting of human values. AB - In the 21st century, psychophysiology will face the challenge of establishing ethical principles and practical means for the genetic and social influencing of the development of human beings. Neuronal imprinting of beliefs and morality within infantile minds will be necessary for the peaceful coexistence of races and cultures. This process requires study and consideration, among others, of the following psychophysiological facts: (1) Genes do not transmit moral values. (2) Material support of physiological activities is necessary for the existence and development of mental functions. (3) Imprinting of human values is based on material changes within neuronal structures. (4) Early neuronal imprinting is performed without personal awareness or consent of the individual and depends on sensory inputs, mainly from the social structure of the group. (5) Biological structures lack values. Personal and social antagonisms do not depend on genes, but on cultural indoctrination. (6) Pleasure and punishment (positive and negative reinforcement) are the two main elements, which regulate animal and human behavior. (7) Values must be chosen by adults, who decide the questions 'why'? 'when'? 'which ones'?, 'who should teach'?, 'what?' and 'how'? (8) Many biological imperatives are shared by all animals and by all people. Human beings may be considered the 'crickets of the Universe', unable to understand the mysteries of nature because of our insufficient neuronal capacity. (9) Our emotional life is mainly related to the structure of the limbic system controlled by the neocortex. (10) New theories based on the integration of physics, chemistry, biology and other specific areas of knowledge, as proposed by the General Theory of Systems, will avoid 'opposites', favoring the acceptance of complementary aspects of reality. (11) Early education will promote preferential learning which depends on both genetic endowment and neuronal development influenced by experience. It is the responsibility of psychophysiology to establish the guidelines for better education, clarifying the material and psychological aspects of the mind. PMID- 10677651 TI - The assessment and management of third party risk around a major airport. AB - Schiphol, the main airport of the Netherlands, is growing rapidly. The aircraft movements, also growing in number, place a considerable environmental burden on the surrounding population, notably, noise and odour nuisance and risks. In the process of deciding on how to extend the capacity of the airport to accommodate the anticipated twofold growth in the number of movements with respect to 1990, environmental problems form a major concern. The concern about risks for the surrounding population was enhanced after the crash on 4 October 1992, in which a Boeing 747 cargo carrier bored into a block of flats in a suburb of Amsterdam near Schiphol. In this accident, the four crew members were killed, together with 39 inhabitants of the flats/apartment building. These risks were studied as part of the Environmental Impact Assessment (EIA). To make these studies useful for decision making necessitated a major improvement in the available techniques for risk quantification. The results of the quantitative analyses, using several different methods, have all indicated that the activities of Schiphol pose a considerable risk compared to other major industrial activities in the Netherlands. This paper describes the development of the methodology from 1990 in the light of the policy context in which it took place. Use of the methods in the decision-making process is illustrated by describing the current status of this process. PMID- 10677652 TI - Integrated safety planning for underground systems. AB - Underground systems are becoming increasingly popular for business activities, transportation systems, and storage purposes. Safety planning for underground systems calls for an integrated approach which considers the interests of many parties, the dynamics of different activities, and the potential threats posed by hazardous materials. A visual interactive modeling approach is presented which helps organizations to derive a safety concept for underground systems. The modeling approach emphasizes procedural aspects of dealing with multiple parties, as well as conceptual and analytic aspects of assessing risks and defining safety goals. The paper summarizes the framework for developing a safety concept for underground systems which was developed for the Dutch Ministry of the Interior. A hypothetical example is discussed to illustrate the theoretical constructs. PMID- 10677653 TI - Application of risk assessment and decision analysis to the evaluation, ranking and selection of environmental remediation alternatives. AB - A single framework integrating risk assessment and decision analysis methods for evaluating, ranking and selecting preferred remediation alternatives at a contaminated site was developed and demonstrated. The methodology used relies on stakeholder inputs throughout the entire process and employs those inputs to combine the results of multiple risk assessments to arrive at a total impact for each remediation alternative. The total impact values allow the ranking of the alternatives, which in turn, serves as the basis for deliberations among the stakeholders in order to identify the preferred alternative. Six major risk or impact categories were considered in the evaluation of the alternatives: human health and safety, environmental protection, life cycle cost, socio-economics, cultural, archeological and historical resources, and programmatic assumptions. PMID- 10677654 TI - Risk analysis and safety policy developments in the Netherlands. AB - In the Netherlands, external safety policy has been developed and implemented since the early eighties on the basis of a risk-based approach involving quantitative criteria for the tolerability of risk. Good experiences have been gained with the risk policy that applies to some 4000 establishments in the Netherlands where hazardous substances are present. On the basis of these experiences, legislation is now being prepared to give the risk tolerability criteria a full legal basis. This is aimed, in particular, to balance between risk control measures at the source through the licensing system, and spatial planning instruments to protect, e.g. residential areas against major hazards. The revision of the Seveso directive (96/82/EC) leads to the implementation of an integrated form of safety reporting, evaluation and inspection. Practical tools were developed for this implementation, e.g. for facilitating the selection of establishments and for assessing risks from major hazard establishments to surface water. In the past few years, the application of risk-based safety policy has been extended to other fields than establishments, e.g. for transport of hazardous chemicals and external safety of airports. PMID- 10677655 TI - Risk management of LPG transport activities in Hong Kong. AB - This paper gives a background to risk management of liquefied petroleum gas (LPG) transport activities, with special regard to the activities taking place in Hong Kong. In particular, it looks at the recent activities undertaken by the Government of the Hong Kong Special Administrative Region (SAR); the recent risk assessment of LPG transport in the Territory, the measures developed to minimise the risks (including risk management improvements) and the risk management activities undertaken by the Government and the operators. PMID- 10677656 TI - Human factors impact on risk analysis of complex systems. AB - This paper discusses the methods and techniques that are applied for including human factors considerations into risk analysis of modern plants. The application of new control design principles and the extensive use of automation have strongly modified the role of operators, who have progressively become supervisors of automatically performed procedures and decision makers in a context of shared management processes. This implies that cognitive functions and organisational factors affect risk analysis much more than behavioural and physical performances. Another crucial issue of human reliability assessment concerns the dynamic nature of human-machine interaction. This feature covers a wide spectrum of real situations, but demands quite complex and extensive data. These considerations favour the development of new and evolutionary techniques which must be confronted with the requirements and needs of different types of risk analysis be carried out for different objectives, such as quantitative risk analysis, safety management, accident investigation, risk-based decision making and risk-based regulations. Advantages and areas of application of different techniques are briefly discussed, without attempting to develop a detailed comparison. PMID- 10677657 TI - 'Worst case' methodology for the initial assessment of societal risk from proposed major accident installations. AB - This paper considers the application of one of the weighted risk indicators used by the Major Hazards Assessment Unit (MHAU) of the Health and Safety Executive (HSE) in formulating advice to local planning authorities on the siting of new major accident hazard installations. In such cases the primary consideration is to ensure that the proposed installation would not be incompatible with existing developments in the vicinity, as identified by the categorisation of the existing developments and the estimation of individual risk values at those developments. In addition a simple methodology, described here, based on MHAU's "Risk Integral" and a single "worst case" even analysis, is used to enable the societal risk aspects of the hazardous installation to be considered at an early stage of the proposal, and to determine the degree of analysis that will be necessary to enable HSE to give appropriate advice. PMID- 10677658 TI - Mitigation of dense gas releases in buildings: use of simple models. AB - When an accidental release of a hazardous material is considered within a safety case or risk assessment, its off-site effects are generally assessed by calculating the dispersion of vapour from the site. Although most installations handling flammables will be in the open air, many types of plant, particularly those handling toxics, are enclosed, partly to provide some form of containment and hence to mitigate the effects of any release. When such a release occurs within a building, the gas or vapour will undergo some mixing before emerging from any openings. The degree of mixing will depend upon the building geometry and the nature of the ventilation, which in turn may be modified by the leak. This situation is considered in this paper, with specific application to calculating the rate of release of a dense vapour from a building. All the calculations presented are based upon simple zone modelling, such that the region occupied by the vapour is assumed to be well mixed, and, in the isothermal case, either its concentration or its depth increases as it is fed by the gas leak. Transfer of air or gas/air mixture through the building openings is estimated by use of standard ventilation calculation methods. For the non-isothermal case, a preliminary model is presented in which it is assumed that there is complete mixing throughout the building and no wind-driven ventilation effects. A moderate release of chlorine is used as an example, and results are shown of the effects of various ventilation possibilities on the release rate to the atmosphere. In addition, comparisons are given between model results and experimental data, demonstrating the level of confidence which can be placed in the models, and also identifying areas where there is scope for further improvement. PMID- 10677659 TI - Consequence analysis in LPG installation using an integrated computer package. AB - This paper presents the prototype of the computer code, Atlantide, developed to assess the consequences associated with accidental events that can occur in a LPG storage plant. The characteristic of Atlantide is to be simple enough but at the same time adequate to cope with consequence analysis as required by Italian legislation in fulfilling the Seveso Directive. The application of Atlantide is appropriate for LPG storage/transferring installations. The models and correlations implemented in the code are relevant to flashing liquid releases, heavy gas dispersion and other typical phenomena such as BLEVE/Fireball. The computer code allows, on the basis of the operating/design characteristics, the study of the relevant accidental events from the evaluation of the release rate (liquid, gaseous and two-phase) in the unit involved, to the analysis of the subsequent evaporation and dispersion, up to the assessment of the final phenomena of fire and explosion. This is done taking as reference simplified Event Trees which describe the evolution of accidental scenarios, taking into account the most likely meteorological conditions, the different release situations and other features typical of a LPG installation. The limited input data required and the automatic linking between the single models, that are activated in a defined sequence, depending on the accidental event selected, minimize both the time required for the risk analysis and the possibility of errors. Models and equations implemented in Atlantide have been selected from public literature or in-house developed software and tailored with the aim to be easy to use and fast to run but, nevertheless, able to provide realistic simulation of the accidental event as well as reliable results, in terms of physical effects and hazardous areas. The results have been compared with those of other internationally recognized codes and with the criteria adopted by Italian authorities to verify the Safety Reports for LPG installations. A brief of the theoretical basis of each model implemented in Atlantide and an example of application are included in the paper. PMID- 10677660 TI - Use of QRA for decision support in the design of an offshore oil production installation. AB - QRA is today widely used as a tool for decision support in the offshore industry. Its use has gradually changed from a prescribed analysis for verification purposes to a tool being actively used in an integrated mode. The paper describes its use in the design of a modern offshore platform. The paper addresses work methodology, selection of tools and data, organisation of QRA with other activities. Specific examples are given. PMID- 10677661 TI - Mitigation of dense gas releases within buildings: validation of CFD modelling. AB - When an accidental release of a hazardous material is considered within a safety case or risk assessment, its off-site effects are generally assessed by calculating the dispersion of vapour from the site. Although most installations handling flammable materials will be in the open air, many types of plant, particularly those handling toxics, are enclosed, partly to provide some form of containment and hence, to mitigate the effects of any release. When such a release occurs within a building, the gas or vapour will undergo some mixing before emerging from any opening. The degree of mixing will depend upon the building geometry and the nature of the ventilation, which in turn may be modified by the leak. This situation is considered in this paper, with specific application to calculating the rate of release of a dense vapour from a building. The paper describes the application of computational fluid dynamics (CFD) techniques to modelling the release and mixing processes within buildings. Examples of validation calculations for simple geometric arrangements, as well as more complex geometries representative of an industrial site, are described. The results demonstrate the capabilities of CFD for this application but highlight the need for careful modelling of the near-wall flows and heat transfer, and need for an accurate fluid dynamics and thermodynamic representation of the release source. PMID- 10677662 TI - Improved nuclear power plant operations and safety through performance-based safety regulation. AB - This paper illustrates some of the promise and needed future work for risk informed, performance-based regulation (RIPBR). RIPBR is an evolving alternative to the current prescriptive method of nuclear safety regulation. Prescriptive regulation effectively constitutes a long, fragmented checklist of requirements that safety-related systems in a plant must satisfy. RIPBR, instead, concentrates upon satisfying negotiated performance goals and incentives for judging and rewarding licensee behavior to improve safety and reduce costs. In a project reported here, a case study was conducted concerning a pressurized water reactor (PWR) emergency diesel generator (EDG). Overall, this work has shown that the methods of RIPBR are feasible to use, and capable of justifying simultaneous safety and economic nuclear power improvements. However, it also reveals several areas where the framework of RIPBR should be strengthened. First, researchers need better data and understanding regarding individual component-failure modes that may cause components to fail. Not only are more data needed on failure rates, but more data and understanding are needed to enable analysts to evaluate whether these failures become more likely as the interval between tests is increased. This is because the current state of failure data is not sufficiently finely detailed to define the failure rates of individual component failure modes; such knowledge is needed when changing component-specific regulatory requirements. Second, the role of component testing, given that a component has failed, needs to be strengthened within the context of RIPBR. This includes formulating requirements for updating the prior probability distribution of a component failure rate and conducting additional or more frequent testing. Finally, as a means of compensating for unavoidable uncertainty as an obstacle to regulatory decision-making, limits to knowledge must be treated explicitly and formally. This treatment includes the formulation of probabilities through expert solicitation and the review of risk-informed, performance-based and engineering analyses used to evaluate proposed changes to existing technical specifications. PMID- 10677663 TI - Statistical analysis of domino chemical accidents. AB - A set of chemical accidents is retrieved from the literature and classified with regard to the substance involved and whether domino effects are present. This set of accidents and each of the classes defined are statistically analyzed with respect to its severity and comparison is made between domino and non-domino accidents. The analysis reveals that each accident category shows characteristic patterns in terms of fatalities caused and domino effects likelihood. Moreover, chemical accidents severity frequencies are described by using a two-parameter, revised form of the Pareto probability density function. The range within which the values of the parameters lie is investigated using Bayesian inference. PMID- 10677664 TI - Uncertainty in compartmental models for hazardous materials - a case study. AB - Performing uncertainty analysis on compartmental models is the main topic of this article. Elements of the methodology developed during a joint CEC/USNRC accident consequence code uncertainty analysis are introduced. The uncertainty is quantified using structured expert judgment. Experts are queried about physically observable quantities. Many code input parameters of the accident consequence codes are not physically observable but are used to predict observable quantities. Therefore, a probabilistic inversion technique was developed which 'transfers' the uncertainty from the physically observable quantities to the code input parameters. The probabilistic inversion technique is illustrated using the compartmental model of systemic retention of Sr in the human body. The article is concluded with a discussion on capturing uncertainty via compartmental models. PMID- 10677665 TI - Standard methods for land-use planning to determine the effects on societal risk. AB - In the Netherlands, the individual risk and the societal risk are used in efforts to reduce the number of people exposed to the effects of an accident. In principle, the societal risk for each new land-use plan should be recalculated. Since this is proving increasingly cumbersome for planning agencies, several methods have been developed for SEVESO establishments and establishments for which in the Netherlands a generic zoning policy is used to determine the effects of new land-use plans on the societal risk. The methods give the uniform population density from a certain distance around the establishment at which the indicative limit for the societal risk is not exceeded. Correction factors are determined for non-uniform population distributions around the establishment, non continuous residence times and deviating societal risk limits. Using these methods allows decision-making without the necessity of repeating quantified risk analyses for each alternative proposal. PMID- 10677666 TI - Hazardous materials transportation: a risk-analysis-based routing methodology. AB - This paper introduces a new methodology based on risk analysis for the selection of the best route for the transport of a hazardous substance. In order to perform this optimisation, the network is considered as a graph composed by nodes and arcs; each arc is assigned a cost per unit vehicle travelling on it and a vehicle capacity. After short discussion about risk measures suitable for linear risk sources, the arc capacities are introduced by comparison between the societal and individual risk measures of each arc with hazardous materials transportation risk criteria; then arc costs are defined in order to take into account both transportation out-of-pocket expenses and risk-related costs. The optimisation problem can thus be formulated as a 'minimum cost flow problem', which consists of determining for a specific hazardous substance the cheapest flow distribution, honouring the arc capacities, from the origin nodes to the destination nodes. The main features of the optimisation procedure, implemented on the computer code OPTIPATH, are presented. Test results about shipments of ammonia are discussed and finally further research developments are proposed. PMID- 10677667 TI - Developments in vapour cloud explosion blast modeling. AB - TNT Equivalency methods are widely used for vapour cloud explosion blast modeling. Presently, however, other types of models are available which do not have the fundamental objections TNT Equivalency models have. TNO Multi-Energy method is increasingly accepted as a more reasonable alternative to be used as a simple and practical method. Computer codes based on computational fluid dynamics (CFD) like AutoReaGas, developed by TNO and Century Dynamics, could be used also in case a more rigorous analysis is required. Application of the Multi-Energy method requires knowledge of two parameters describing the explosion: a charge size and a charge strength. During the last years, research has led to an improved determination of the charge strength (i.e., the class number or source overpressure) to be chosen to apply the blast charts. A correlation has been derived relating the charge strength to a set of parameters describing the boundary conditions of the flammable cloud and the fuel in the cloud. A simple approach may not be satisfactory in all situations. The overpressure distribution inside a vapour cloud explosion is generally not homogeneous and the presence of obstructions causes directional blast propagation in the near field. A CFD approach, in which the actual situation is modeled, supplies case-specific results. An overview of the key aspects relevant to the application of the Multi Energy method and CFD modeling is provided. Then the application of the two methods is demonstrated for an example problem involving the calculation of the explosion blast load on a structure at some distance from the explosion in an offshore platform complex. PMID- 10677668 TI - Decision support in nuclear emergencies. AB - In a nuclear emergency, protective actions such as evacuation, sheltering and food bans can be taken to mitigate the consequences of any release of radioactivity. Within the RODOS project, an evaluation framework has been developed to support the assessment of the costs and benefits of potential actions. In order to help the decision makers gain insight into the decision problem and clarify their preferences, guidance can be given in three stages. First, the search of feasible portfolios of protective actions is seen as a constraint satisfaction problem; only those portfolios that satisfy constraints depending on factors such as feasibility are worth further evaluation. Second, the portfolios are ranked based on their consequences and the preferences of the decision makers using either a multi-attribute value or utility function. Third, a natural language report explaining the ranking is produced to help the decision makers gain insight into the decision problem and refine the decision parameters. An intelligent decision system has been developed to demonstrate the feasibility of the framework. PMID- 10677669 TI - Supporting decision makers in land use planning around chemical sites. Case study: expansion of an oil refinery. AB - An approach for supporting decisions on land use around chemical sites - along with a software decision support system (DSS) - based on multi-criteria decision analysis (MCDA; and particularly on the establishment of the set of efficient solutions and letting the final selection depend on local procedures and value tradeoffs) is being illustrated through a case study where five alternative expansions of a refinery are considered along with the existing situation. Alternative land use plans are based on combinations of alternative uses of specific land cells coupled with alternative expansion options. Criteria for evaluating alternative land use plans are the potential loss of life (PLL), the noise levels and the economic benefit resulting for each specific land use plan. PMID- 10677670 TI - Risk informed resource allocation policy: safety can save costs. AB - During economic doldrums, decision making on investments for safety is even more difficult than it already is when funds are abundant. This paper attempts to offer some guidance. After stating the present challenge to prevention of losses in the process industries, the systematic approach of quantified risk assessment is briefly reviewed and improvements in the methodology are mentioned. In addition, attention is given to the use of a risk matrix to survey a plant and to derive a plan of action. Subsequently, the reduction of risk is reviewed. Measures for prevention, protection, and mitigation are discussed. The organization of safety has become at least as important as technical safety of equipment and standards. It is reflected in the introduction of a safety management system. Furthermore, the design process in a pro-active approach is described and the concept of inherent safety is briefly addressed. The concept of Layer of Protection Analysis is explained and also the reason why it is relevant to provide a cost-benefit analysis. Finally, after comments regarding the cost of accidents, the basics of costing and profitability are summarized and a way is suggested to apply this approach to risk-reducing measures. An example is provided on how a selection can be made from a number of alternatives. PMID- 10677671 TI - Risk of hydrocyanic acid release in the electroplating industry. AB - This paper suggests assessing the consequences of hydrocyanic acid (HCN) release into the air by aqueous cyanide solutions in abnormal situations such as the accidental introduction of an acid, or the insertion of a cyanide in a pickling bath. It provides a well-defined source model and its resolution by methods peculiar to mass transport phenomena. The procedure consists of four stages: calculation of the liquid phase concentration, estimate of the HCN liquid-vapour equilibrium, determination of the mass transfer coefficient at the liquid-vapour interface, evaluation of the air concentration of HCN and of the damage distances. The results show that small baths operating at high temperatures are the major sources of risk. The building up of lethal air concentrations, on the other hand, is governed by the values of the mass transfer coefficient, which is itself determined by the flow dynamics and bath geometry. Concerning the magnitude of the risk, the fallout for external emergency planning is slight in all the cases investigated. PMID- 10677672 TI - Off-site ignition probability of flammable gases. AB - A key step in the assessment of risk for installations where flammable liquids or gases are stored is the estimation of ignition probability. A review of current modelling and data confirmed that ignition probability values used in risk analyses tend to be based on extrapolation of limited incident data or, in many cases, on the judgement of those conducting the safety assessment. Existing models tend to assume that ignition probability is a function of release rate (or flammable gas cloud size) alone and they do not consider location, density or type of ignition source. An alternative mathematical framework for calculating ignition probability is outlined in which the approach used is to model the distribution of likely ignition sources and to calculate ignition probability by considering whether the flammable gas cloud will reach these sources. Data are collated on the properties of ignition sources within three generic land-use types: industrial, urban and rural. These data are then incorporated into a working model for ignition probability in a form capable of being implemented within risk analysis models. The sensitivity of the model results to assumptions made in deriving the ignition source properties is discussed and the model is compared with other available ignition probability methods. PMID- 10677673 TI - Through ARIPAR-GIS the quantified area risk analysis supports land-use planning activities. AB - The paper first summarises the main aspects of the ARIPAR methodology whose steps can be applied to quantify the impact on a territory of major accident risks due to processing, storing and transporting dangerous substances. Then the capabilities of the new decision support tool ARIPAR-GIS, implementing the mentioned procedure, are described, together with its main features and types of results. These are clearly shown through a short description of the updated ARIPAR study (reference year 1994), in which the impact of changes due to industrial and transportation dynamics on the Ravenna territory in Italy were evaluated. The brief explanation of how results have been used by local administrations offers the opportunity to discuss about advantages of the quantitative area risk analysis tool in supporting activities of risk management, risk control and land-use planning. PMID- 10677674 TI - PROTEUS, a technical and management model for aquatic risk assessment of industrial spills. AB - The assessment of risks to the aquatic environment related to industrial installations is a priority in environmental pollution control in the Netherlands. Major accidents to the surface water such as the Sandoz incident, but also the high number of smaller accidents that occur every year has invoked the need for an effective method to assess these risks. Two different models have been used in this field in the Netherlands over several years. These two software applications, VERIS and RISAM were developed from two different perspectives: VERIS from the perspective of supplying major accidents related information in the safety report, RISAM form the perspective of controlling risks for both smaller and larger facilities that may pollute surface waters through accidents. Both systems comprised particular strong points: VERIS considers safety management aspects in the assessment, RISAM considers differences in surface water vulnerability and involves quantitative probabilities in the assessment. It was decided to integrate both methods and maintain these strong points in the resulting method. This paper describes the new integrated risk assessment method that now has been developed in a concerted effort between the Ministry of Transport, Public Works and Water Management, the Ministry of Housing, Spatial Planning and Environment, and the National Institute for Public Health and Environment. It also describes the essential elements of the computer program PROTEUS that is based on the new method and that makes the assessment of aquatic risks for industrial activities an easy task, partly due to the automatic generation of the assessment report. PMID- 10677675 TI - Performance-based standards: safety instrumented functions and safety integrity levels. AB - This paper discusses two international performance-based standards, ANSI/ISA S84.01 and IEC d61508 and the requirements they place upon companies that rely on electrical, electronic and programmable electronic systems to perform safety functions. Performance-based regulations are also discussed and common safety elements between the standards and regulations are identified. Several risk analysis techniques that can be used to comply with the aforementioned requirements are discussed and a simple example is used to illustrate the use, advantages and disadvantages of the techniques. The evaluation of safety integrity level (SIL) of the Safety Instrumented System (SIS) in terms of the probability to fail to function is outside the scope of this paper. PMID- 10677676 TI - Risk based methodology for safety improvements in ports. AB - With the introduction of the Formal Safety Assessment in the International Maritime Organisation decision making process regarding new regulations, and the recent tanker disasters resulting in extensive oil pollution, the public and political pressure to improve safety in ports and the shipping industry has increased. Considering that some kind of Safety Report (case) regulations related to marine operations have not been established, and that the ports and shipping industry are at the onset of safety regimes utilised in other industries, a step wise methodology for safety improvements in ports has been developed. In the first step, the hazard identification and the qualitative risk assessment is carried out to establish hazard barriers which are or should be in place to prevent hazards from being released; the controls for managing these hazards are then developed and integrated into the Safety Management System (SMS). In the second and optional step, the areas of high risk are investigated in detail and the approach for risk quantification discussed. The use of the quantitative risk assessment results is illustrated in two examples. PMID- 10677677 TI - Maintenance-based strategies for environmental risk minimization in the process industries. AB - Industry, environmental agencies and the scientific community have all emphasized the need to include environmental impact considerations next to profitability objectives in the design phase of modern chemical processes, responding to the increasing social concern over environmental degradation in the past years. Most environmental impact assessment and minimization approaches, however, are rather qualitative, providing general guidelines. In this work, to overcome their limitations and rigorously represent the defining elements of environmental risk, i.e. the mechanism of occurrence of unexpected events usually related to equipment failure and the severity of their consequences, detailed process, reliability and maintenance characteristics are incorporated within a process optimization framework. The objective concerns the optimization of overall process performance defined as a system effectiveness vector characterized by both the environmental and the profitability functions of the system. Implementation of the framework on a process example identifies the optimal combination of process design and operation as well as preventive maintenance strategies that accomplish the conflicting environmental and profitability targets and quantifies the existing trade-offs between them. PMID- 10677678 TI - Methodological framework for developing decision support systems (DSS) for hazardous materials emergency response operations. AB - The production, storage, and transportation of hazardous materials are processes of vital economic importance for any advanced and technologically complex society. Although the production and distribution of hazardous materials is associated with economic development, there is a significant potential danger to the natural and social environment in the event of their accidental release, a fact that prompts for the development and implementation of methods and techniques that aim to improve hazardous materials risk management decisions. The objective of this paper is to present a unified framework for developing a Decision Support System (DSS) for supporting a vital function of risk management, namely the management of emergency response operations. The proposed framework recognizes the peculiarities of the hazardous materials decision-making environment which is characterized by: (i) multiple stakeholders, i.e., persons and organizations involved in and affected by hazardous materials risk management decisions; (ii) lack of a formal management structure for monitoring and controlling in a unified manner all Emergency Response Resources; (iii) lack of clear distinction and fragmentation of responsibilities of the actors involved in risk management operations; and (iv) dynamic/real-time decisions, i.e., risk determinants change over time. The proposed framework was used in order to develop a DSS for managing emergency response operations for large scale industrial accidents in Western Attica, Greece. PMID- 10677679 TI - XRCC1 keeps DNA from getting stranded. PMID- 10677680 TI - UV lesions located on the leading strand inhibit DNA replication but do not inhibit SV40 T-antigen helicase activity. AB - DNA replication in eucaryotic cells involves a variety of proteins which synthesize the leading and lagging strands in an asymmetric coordinated manner. To analyse the effect of this asymmetry on the translesion synthesis of UV induced lesions, we have incubated SV40 origin-containing plasmids with a unique site-specific cis, syn-cyclobutane dimer or a pyrimidine-pyrimidone (6-4) photoproduct on either the leading or lagging strand template with DNA replication-competent extracts made from human HeLa cells. Two dimensional agarose gel electrophoresis analyses revealed a strong blockage of fork progression only when the UV lesion is located on the leading strand template. Because DNA helicases are responsible for unwinding duplex DNA ahead of the fork and are then the first component to encounter any potential lesion, we tested the effect of these single photoproducts on the unwinding activity of the SV40 T antigen, the major helicase in our in vitro replication assay. We showed that the activity of the SV40 T-antigen helicase is not inhibited by UV-induced DNA lesions in double-stranded DNA substrate. PMID- 10677681 TI - Cellular and molecular effects of bleomycin are modulated by heat shock in Saccharomyces cerevisiae. AB - To study some mechanisms underlying the stress responses in eukaryotic cells, we investigated the effect of heat shock (HS) on the induction of DNA double strand breaks as well as on potentially lethal and mutagenic events induced by the radiomimetic antibiotic bleomycin (BLM) in Saccharomyces cerevisiae. Haploid wild type yeast cells in the logarithmic phase of growth were exposed to different concentrations of BLM (0-30 microg/ml, 1.5 h) without and with a previous HS (38 degrees C, 1 h). Immediately after treatments, survival as well as mutation frequency were determined, and quantitative analysis of chromosomal DNA by laser densitometry were performed both immediately after treatments and after incubation of cells during different time intervals in liquid nutrient medium free of BLM. Our results indicate that HS induces resistance to potentially lethal and mutagenic effects of BLM. Quantitative analysis of chromosomal DNA performed immediately after treatments showed the same DNA fragmentation, either upon BLM as single agent or preceded by HS. However, HS pretreated cells incubated during 4 h in liquid nutrient medium free of BLM repaired DNA double strand breaks more efficiently as compared to non-pretreated cells. On this basis, we propose that the observed HS-induced resistance to BLM depends on a regulatory network acting after DNA-induced damage, which includes genes involved in DNA repair, HS response and DNA metabolism. PMID- 10677683 TI - Mitotic viability and metabolic competence in UV-irradiated yeast cells. AB - Colony formation is the classic method for measuring survival of yeast cells. This method measures mitotic viability and can underestimate the fraction of cells capable of carrying out other DNA processing events. Here, we report an alternative method, based on cell metabolism, to determine the fraction of surviving cells after ultraviolet (UV) irradiation. The reduction of 2,3,5 triphenyl tetrazolium chloride (or TTC) to formazan in mitochondria was compared with cell colony formation and DNA repair capacity in wt cells and two repair deficient strains (rad1Delta and rad7Delta). Both TTC reduction and cell colony formation gave a linear response with different ratios of mitotically viable cells and heat-inactivated cells. However, monitoring the formation of formazan in non-dividing yeast cells that are partially (rad7Delta) or totally (wt) proficient at DNA repair is a more accurate measure of cell survival after UV irradiation. Before repair of UV photoproducts (cis-syn cyclobutane pyrimidine dimers or CPDs) is complete, these two assays give very different results, implying that many damaged cells are metabolically competent but cannot replicate. For example, only 25% of the rad7Delta cells are mitotically viable after a UV dose of 12 J/m(2)75% of these cells are metabolically competent and remove over 55% of the CPDs from their genomic DNA. Moreover, repair of CPDs in wt cells dramatically decreases after the first few hours of liquid holding (L.H.; incubation in water) and correlates with a substantial decrease in cell metabolism over the same time period. In contrast, cell colony formation may be the more accurate indicator of cell survival after UV irradiation of rad1Delta cells (i.e., cells with little DNA repair activity). These results indicate that the metabolic competence of UV-irradiated, non-dividing yeast cells is a much better indicator of cell survival than mitotic viability in partially (or totally) repair proficient yeast cultures. PMID- 10677682 TI - AP lyases and dRPases: commonality of mechanism. AB - Enzymes that release 5'-deoxyribose-5-phosphate (dRP) residues from preincised apurinic/apyrimidinic (AP) DNA have been collectively termed DNA deoxyribophosphodiesterases (dRPases), but they fall into two distinct categories: the hydrolytic dRPases and AP lyases. In order to resolve a number of conflicting reports in the dRPase literature, we examined two putative hydrolytic dRPases (Escherichia coli exonuclease I (exo I) and RecJ) and four AP lyases (E. coli 2, 6-dihydroxy-5N-formamidopyrimidine (Fapy) DNA glycosylase (Fpg) and endonuclease III (endo III), bacteriophage T4 endonuclease V (endo V), and rat polymerase beta (beta-pol)) for their abilities to (i) excise dRP from preincised AP DNA and (ii) incise AP DNA. Although exo I and RecJ exhibited robust 3' to 5' and 5' to 3' exonucleolytic activities, respectively, on appropriate substrates, they failed to demonstrate detectable dRPase activity. All four AP lyases possessed both dRPase and traditional AP lyase activities, albeit to varying degrees. Moreover, as best illustrated with Fpg, AP lyase enzymes could be trapped on both preincised and unincised AP DNA using NaBH(4) as the reducing agent. These results further support the assertion that the catalytic mechanism of the AP lyases, the beta-elimination reaction, does proceed through an imine enzyme-DNA intermediate and that the active site residues responsible for dRP release must contain primary amines. Further, these data indicate a biological significance for the beta-elimination reaction of DNA glycosylase/AP lyases in that they, in concert with hydrolytic AP endonucleases, can create appropriate gapped substrates for short patch base excision repair (BER) synthesis to occur efficiently. PMID- 10677684 TI - Overexpression of bacterial RecA protein stimulates homologous recombination in somatic mammalian cells. AB - The pairing of homologous molecules and strand exchange is a key event in homologous recombination promoted by RecA protein in Escherichia coli. Structural homologs of RecA are widely distributed in eukaryotes including mouse and man. As has been shown, human HsRad51 protein is not only structural but also functional homolog of RecA. The question arises whether the bacterial functional homolog of Rad51 can function in mammalian cells and increase the frequency of the homologous recombination. To investigate possible effects of bacterial RecA protein on the frequency of homologous recombination in mammalian cells, the E. coli RecA protein fused with a nuclear location signal from the large T antigen of simian virus 40 was overexpressed in the mouse F9 teratocarcinoma cells. We found that the frequency of gene targeting at the hprt locus was 10-fold increased in the mouse cells expressing the nucleus-targeted RecA protein. Southern blot analysis of individual clones that were generated by targeting recombination revealed predicted type of alterations in hprt gene. The data indicate that the bacterial nucleus-targeted RecA protein can stimulate homologous recombination in mammalian cells. PMID- 10677685 TI - Chronic lymphocytic leukemia lymphocytes lack the capacity to repair UVC-induced lesions. AB - Cells from chronic lymphocytic leukemia (CLL) patients and from healthy individuals were irradiated with UVC and incubated for varying periods of time. The number of single strand breaks and alkali-labile sites was determined by comet analysis. Unirradiated CLL and healthy cells exhibited no significant numbers of single strand breaks. The extent of DNA damage was found to increase with dose for both healthy and CLL cells. However, the CLL cells had much more extensive DNA fragmentation than healthy cells at each dose. Deoxyribonucleoside supplemented medium inhibited comet formation in both cell types. Thymidine alone produced the same effect. In healthy cells, repair of lesions was complete after 4 h of incubation as indicated by the absence of comet formation. The CLL cells exhibited no significant repair even after 48 h. CLL lymphocytes are killed by very low doses of UVC radiation. The results reported here suggest that this hypersensitivity results from the inability of CLL cells to repair UVC-induced DNA damage and a contributing factor is the low amounts of intracellular deoxyribonucleosides. PMID- 10677686 TI - Dominant sensitization variants of human O(6)-methylguanine-DNA-methyltransferase obtained by a mutational screen of surface residues. AB - A scanning mutagenesis experiment was performed on human O(6)-methylguanine methyltransferase (hMGMT), directed largely at non-conserved surface residues that have not previously been studied. Variants typically contained two or more substitutions. Two of the 16 variants characterized in detail are inactive for methyltransfer, but increase the cytotoxicity and mutagenic effects of methylating agents. This phenotype is reminiscent of a variant (C145A) that has a mutation in the methyl-accepting cysteine. C145A is inactive, but reportedly binds methylated DNA and confers sensitivity to methylating agents. The sensitization phenotype of the two new variants is more striking in strains that are wild-type for DNA repair than in strains that are deficient for repair, suggesting that these proteins inhibit functional DNA repair proteins by competitively binding to methylated DNA. Both variants have multiple substitutions in the last helix of the protein. These results suggest that the C terminal helix is necessary for methyltransfer activity, but not for methylguanine-specific binding. PMID- 10677687 TI - Foreword PMID- 10677688 TI - Chagas disease, from discovery to control - and beyond: history, myths and lessons to take home. PMID- 10677689 TI - A short review on the morphology of Trypanosoma cruzi: from 1909 to 1999. AB - The morphology of Trypanosoma cruzi is reviewed since the initial description of Giemsa-stained preparations by Carlos Chagas until the most recent micrographs obtained with freeze-fracture techniques. Special emphasis is given to structures such as the cell surface, the flagellum, the kinetoplast, the reservosomes and the endocytic pathway, and the acidocalcisomes. PMID- 10677690 TI - Relevant glycoconjugates on the surface of Trypanosoma cruzi. PMID- 10677691 TI - A role for extracellular amastigotes in the immunopathology of Chagas disease. AB - In spite of the growing knowledge obtained about immune control of Trypanosoma cruzi infection, the mechanisms responsible for the variable clinico-pathological expression of Chagas disease remain unknown. In a twist from previous concepts, recent studies indicated that tissue parasitism is a pre-requisite for the development of chronic myocarditis. This fundamental concept, together with the realization that T. cruzi organisms consist of genetically heterogeneous clones, offers a new framework for studies of molecular pathogenesis. In the present article, we will discuss in general terms the possible implications of genetic variability of T. cruzi antigens and proteases to immunopathology. Peptide epitopes from a highly polymorphic subfamily of trans-sialidase (TS) antigens were recently identified as targets of killer T cell (CTL) responses, both in mice and humans. While some class I MHC restricted CTL recognize epitopes derived from amastigote-specific TS-related antigens (TSRA), others are targeted to peptide epitopes originating from trypomastigote-specific TSRA. A mechanistic hypothesis is proposed to explain how the functional activity and specificity of class I MHC restricted killer T cells may control the extent to which tissue are exposed to prematurely released amastigotes. Chronic immunopathology may be exacerbated due the progressive accumulation of amastigote-derived antigens and pro-inflammatory molecules (eg. GPI-mucins and kinin-releasing proteases) in dead macrophage bodies. PMID- 10677692 TI - The population structure of Trypanosoma cruzi: expanded analysis of 54 strains using eight polymorphic CA-repeat microsatellites. AB - Recently we cloned and sequenced the first eight Trypanosoma cruzi polymorphic microsatellite loci and studied 31 clones and strains to obtain valuable information about the population structure of the parasite. We have now studied 23 further strains, increasing from 11 to 31 the number of strains obtained from patients with chronic Chagas disease. This expanded set of 54 strains and clones analyzed with the eight microsatellites markers confirmed the previously observed diploidy, clonal population organization and very high polymorphism of T. cruzi. Moreover, this new study disclosed two new features of the population genetic structure of T. cruzi. The first was the discovery that, similarly to what we had previously shown for strains isolated from insect vectors, mammals and humans with acute disease, isolates from patients in the chronic phase of Chagas disease could also be multiclonal, albeit at a reduced proportion. Second, when we used parsimony to display the genetic relationship among the clonal lineages in an unrooted Wagner network we observed, like before, a good correlation of the tree topography with the classification in three clusters on the basis of single locus analysis of the ribosomal RNA genes. However, a significant new finding was that now the strains belonging to cluster 2 split in two distant sub-clusters. This observation suggests that the evolutionary history of T. cruzi may be more complex than we previously thought. PMID- 10677693 TI - Immunopathology of Chagas disease. AB - The main clinical forms of Chagas disease (acute, indeterminate and chronic cardiac) present strong evidences for the participation of the immune system on pathogenesis. Although parasite multiplication is evident during acute infection, the intense acute myocarditis of this phase exhibits clear ultrastructural signs of cell-mediated immune damage, inflicted to parasitized and non-parasitized myocardiocytes and to the endothelium of myocardial capillaries (microangiopathy). Inflammation subsides almost completely when immunity decreases parasite load and suppressor factors modulate host reaction, but inflammation does not disappear when the disease enters the indeterminate phase. Inflammation becomes mild and focal and undergoes cyclic changes leading to complete resolution. However, the process is maintained because the disappearance of old focal lesions is balanced by the upsurge of new ones. This equilibrium allows for prolonged host survival in the absence of symptoms or signs of disease. The chronic cardiac form is represented by a delayed-type, cell-mediated diffuse myocarditis, that probably ensues when the suppressive mechanisms, operative during the indeterminate phase, become defaulted. The mechanism responsible for the transition from the indeterminate to the cardiac form, is poorly understood. PMID- 10677694 TI - Evolution of the clinical and epidemiological knowledge about Chagas disease 90 years after its discovery. AB - Three different periods may be considered in the evolution of knowledge about the clinical and epidemiological aspects of Chagas disease since its discovery: (a) early period concerning the studies carried out by Carlos Chagas in Lassance with the collaboration of other investigators of the Manguinhos School. At that time the disease was described and the parasite, transmitters and reservoirs were studied. The coexistence of endemic goiter in the same region generated some confusion about the clinical forms of the disease; (b) second period involving uncertainty and the description of isolated cases, which lasted until the 1940 decade. Many acute cases were described during this period and the disease was recognized in many Latin American countries. Particularly important were the studies of the Argentine Mission of Regional Pathology Studies, which culminated with the description of the Romana sign in the 1930 decade, facilitating the diagnosis of the early phase of the disease. However, the chronic phase, which was the most important, continued to be difficult to recognize; (c) period of consolidation of knowledge and recognition of the importance of Chagas disease. Studies conducted by Laranja, Dias and Nobrega in Bambui updated the description of Chagas heart disease made by Carlos Chagas and Eurico Villela. From then on, the disease was more easily recognized, especially with the emphasis on the use of a serologic diagnosis; (d) period of enlargement of knowledges on the disease. The studies on denervation conducted in Ribeirao Preto by Fritz Koberle starting in the 1950 decade led to a better understanding of the relations between Chagas disease and megaesophagus and other visceral megas detected in endemic areas. PMID- 10677695 TI - [Evolution of the knowledge about Chagas disease vectors 90 years after its discovery]. PMID- 10677696 TI - Prevention of transfusional Trypanosoma cruzi infection in Latin America. AB - Trypanosoma cruzi is a protozoan infection widely spread in Latin America, from Mexico in the north to Argentina and Chile in the south. The second most important way of acquiring the infection is by blood transfusion. Even if most countries of Latin America have law/decree/norms, that make mandatory the screening of blood donors for infectious diseases, including T. cruzi (El Salvador and Nicaragua do not have laws on the subject), there is usually no enforcement or it is very lax. Analysis of published serologic surveys of T. cruzi antibodies in blood donors done in 1993, indicating the number of donors and screening coverage for T. cruzi in ten countries of Central and South America indicated that the probability of receiving a potentially infected transfusion unit in each country varied from 1,096 per 10,000 transfusions in Bolivia, the highest, to 13.02 or 13.86 per 10,000 transfusions in Honduras and Venezuela respectively, where screening coverage was 100%. On the other hand the probability of transmitting a T. cruzi infected unit was 219/10,000 in Bolivia, 24/10,000 in Colombia, 17/10,000 in El Salvador, and around 2-12/10,000 for the seven other countries. Infectivity risks defined as the likelihood of being infected when receiving an infected transfusion unit were assumed to be 20% for T. cruzi. Based on this, estimates of the absolute number of infections induced by transfusion indicated that they were 832, 236, and 875 in Bolivia, Chile and Colombia respectively. In all the other countries varied from seven in Honduras to 85 in El Salvador. Since 1993, the situation has improved. At that time only Honduras and Venezuela screened 100% of donors, while seven countries, Argentina, Colombia, El Salvador, Honduras, Paraguay, Uruguay and Venezuela, did the same in 1996. In Central America, without information from Guatemala, the screening of donors for T. cruzi prevented the transfusion of 1,481 infected units and the potential infection of 300 individuals in 1996. In the same year, in seven countries of South America, the screening prevented the transfusion of 36,017 infected units and 7, 201 potential cases of transfusional infection. PMID- 10677697 TI - The evolution of Chagas disease (American trypanosomiasis) control after 90 years since Carlos Chagas discovery. PMID- 10677698 TI - The noble enigma: Chagas' nominations for the Nobel prize. AB - Carlos Chagas, a Brazilian physician, discovered the American trypanosomiasis in 1909. Like other remarkable discoveries of those days, his work helped to articulate the insect-vector theory and other theoretical guidelines in tropical medicine. Unlike all other discoveries, all the stages of this work were accomplished in a few months and by a single man. Chagas' discovery was widely recognized at home and abroad. He was twice nominated for the Nobel Prize - in 1913 and in 1921-, but never received the award. Evidence suggests that the reasons for this failure are related to the violent opposition that Chagas faced in Brazil. The contentions towards Chagas were related to a rejection of the meritocratic procedures that gave him prominence, as well as to local petty politics. PMID- 10677699 TI - Biology and ultra-structure of Trypanosoma cruzi: a 90-year old challenge for scientists. PMID- 10677700 TI - Features of host cell invasion by different infective forms of Trypanosoma cruzi. AB - Through its life cycle from the insect vector to mammalian hosts Trypanosoma cruzi has developed clever strategies to reach the intracellular milieu where it grows sheltered from the hosts' immune system. We have been interested in several aspects of in vitro interactions of different infective forms of the parasite with cultured mammalian cells. We have observed that not only the classically infective trypomastigotes but also amastigotes, originated from the extracellular differentiation of trypomastigotes, can infect cultured cells. Interestingly, the process of invasion of different parasite infective forms is remarkably distinct and also highly dependent on the host cell type. PMID- 10677701 TI - The reservosome of Trypanosoma cruzi epimastigotes: an organelle of the endocytic pathway with a role on metacyclogenesis. PMID- 10677702 TI - Internalization of components of the host cell plasma membrane during infection by Trypanosoma cruzi. AB - Epimastigote and trypomastigote forms of Trypanosoma cruzi attach to the macrophage surface and are internalized with the formation of a membrane bounded vacuole, known as the parasitophorous vacuole (PV). In order to determine if components of the host cell membrane are internalized during formation of the PV we labeled the macrophage surface with fluorescent probes for proteins, lipids and sialic acid residues and then allowed the labeled cells to interact with the parasites. The interaction process was interrupted after 1 hr at 37 masculineC and the distribution of the probes analyzed by confocal laser scanning microscopy. During attachment of the parasites to the macrophage surface an intense labeling of the attachment regions was observed. Subsequently labeling of the membrane lining the parasitophorous vacuole containing epimastigote and trypomastigote forms was seen. Labeling was not uniform, with regions of intense and light or no labeling. The results obtained show that host cell membrane lipids, proteins and sialoglycoconjugates contribute to the formation of the membrane lining the PV containing epimastigote and trypomastigote T. cruzi forms. Lysosomes of the host cell may participate in the process of PV membrane formation. PMID- 10677703 TI - Trypanosoma cruzi-cardiomyocytes: new contributions regarding a better understanding of this interaction. AB - The present paper summarizes new approaches regarding the progress done to the understanding of the interaction of Trypanosoma cruzi-cardiomyocytes. Mannose receptors localized at the surface of heart muscle cell are involved in binding and uptake of the parasite. One of the most striking events in the parasite-heart muscle cells interaction is the disruption of the actin cytoskeleton. We have investigated the regulation of the actin mRNA during the cytopathology induced in myocardial cells by the parasite. T. cruzi invasion increases calcium resting levels in cardiomyocytes. We have previously shown that Ca2+ ATPase of the sarcoplasmic reticulum (SERCA) is involved in the invasion of T. cruzi in cardiomyocytes. Treating the cells with thapsigargin, a drug that binds to all SERCA ATPases and causes depletion of intracellular calcium stores, we found a 75% inhibition in the T. cruzi-cardiomyocytes invasion. PMID- 10677704 TI - Why studies on invasion of host cell by Trypanosoma cruzi using established cell lines or primary cell cultures give conflicting results? PMID- 10677705 TI - Recent issues of the biochemistry and molecular biology of Trypanosoma cruzi. PMID- 10677706 TI - Epidemiology, biochemistry and evolution of Trypanosoma cruzi lineages based on ribosomal RNA sequences. PMID- 10677707 TI - Differential gene expression during Trypanosoma cruzi metacyclogenesis. AB - The transformation of epimastigotes into metacyclic trypomastigotes involves changes in the pattern of expressed genes, resulting in important morphological and functional differences between these developmental forms of Trypanosoma cruzi. In order to identify and characterize genes involved in triggering the metacyclogenesis process and in conferring to metacyclic trypomastigotes their stage specific biological properties, we have developed a method allowing the isolation of genes specifically expressed when comparing two close related cell populations (representation of differential expression or RDE). The method is based on the PCR amplification of gene sequences selected by hybridizing and subtracting the populations in such a way that after some cycles of hybridization amplification genes specific to a given population are highly enriched. The use of this method in the analysis of differential gene expression during T. cruzi metacyclogenesis (6 hr and 24 hr of differentiation and metacyclic trypomastigotes) resulted in the isolation of several clones from each time point. Northern blot analysis showed that some genes are transiently expressed (6 hr and 24 hr differentiating cells), while others are present in differentiating cells and in metacyclic trypomastigotes. Nucleotide sequencing of six clones characterized so far showed that they do not display any homology to gene sequences available in the GeneBank. PMID- 10677708 TI - Organization and expression of a multigene family encoding the surface glycoproteins of Trypanosoma cruzi metacyclic trypomastigotes involved in the cell invasion. PMID- 10677709 TI - Trypanosoma cruzi mucins: potential functions of a complex structure. PMID- 10677710 TI - Characterization of Trypanosoma cruzi. PMID- 10677711 TI - Taxonomy of Trypanosoma cruzi: a commentary on characterization and nomenclature. PMID- 10677712 TI - Trypanosoma cruzi: clonal structure of parasite strains and the importance of principal clones. PMID- 10677713 TI - Genetic diversity and genetic exchange in Trypanosoma cruzi: dual drug-resistant "progeny" from episomal transformants. AB - Extensive characterisation of Trypanosoma cruzi by isoenzyme phenotypes has separated the species into three principal zymodeme groups, Z1, Z2 and Z3, and into many individual zymodemes. There is marked diversity within Z2. A strong correlation has been demonstrated between the strain clusters determined by isoenzymes and those obtained using random amplified polymorphic DNA (RAPD) profiles. Polymorphisms in ribosomal RNA genes, in mini-exon genes, and microsatellite fingerprinting indicate the presence of at least two principal T. cruzi genetic lineages. Lineage 1 appears to correspond with Z2 and lineage 2 with Z1. Z1 (lineage 2) is associated with Didelphis. Z2 (lineage 1) may be associated with a primate host. Departures from Hardy-Weinberg equilibrium and linkage disequilibrium indicate that propagation of T. cruzi is predominantly clonal. Nevertheless, two studies show putative homozygotes and heterozygotes circulating sympatrically: the allozyme frequencies for phosphoglucomutase, and hybrid RAPD profiles suggest that genetic exchange may be a current phenomenon in some T. cruzi transmission cycles. We were able to isolate dual drug-resistant T. cruzi biological clones following copassage of putative parents carrying single episomal drug-resistant markers. A multiplex PCR confirmed that dual drug resistant clones carried both episomal plasmids. Preliminary karyotype analysis suggests that recombination may not be confined to the extranuclear genome. PMID- 10677714 TI - Populational heterogeneity of Brazilian Trypanosoma cruzi isolates revealed by the mini-exon and ribosomal spacers. PMID- 10677715 TI - Reflections on the population dynamics of Trypanosoma cruzi: heterogeneity versus plasticity. PMID- 10677716 TI - The sylvatic cycle of Trypanosoma cruzi: a still unsolved puzzle. PMID- 10677717 TI - Morphobiological characterization of Trypanosoma cruzi Chagas, 1909 and its distinction from other trypanosomes. PMID- 10677718 TI - Trypanosoma cruzi - vector-vertebrate hosts interactions. PMID- 10677719 TI - Biological factors involving Trypanosoma cruzi life cycle in the invertebrate vector, Rhodnius prolixus. PMID- 10677720 TI - Some morphological and molecular aspects of the life cycle of Trypansoma cruzi in the insect vector. PMID- 10677721 TI - Immunity in Rhodnius prolixus: trypanosomatid-vector interactions. PMID- 10677722 TI - Triatominae as a model of morphological plasticity under ecological pressure. AB - The use of biochemical and genetic characters to explore species or population relationships has been applied to taxonomic questions since the 60s. In responding to the central question of the evolutionary history of Triatominae, i.e. their monophyletic or polyphyletic origin, two important questions arise (i) to what extent is the morphologically-based classification valid for assessing phylogenetic relationships? and (ii) what are the main mechanisms underlying speciation in Triatominae? Phenetic and genetic studies so far developed suggest that speciation in Triatominae may be a rapid process mainly driven by ecological factors. PMID- 10677724 TI - The synanthropic process of Chagas disease vectors in Brazil, with special attention to Triatoma brasiliensis Neiva, 1911 (Hemiptera, reduviidae, triatominae) population, genetical, ecological, and epidemiological aspects. PMID- 10677723 TI - Mitochondrial DNA variation of Triatoma infestans populations and its implication on the specific status of T. melanosoma. AB - DNA sequence comparison of 412 base-pairs fragments of the mitochondrial cytochrome B gene was used to infer the genetic structure of nine geographical Triatoma infestans populations and their phylogenetic relationship with T. melanosoma and T. brasiliensis. T. infestans and T. melanosoma were compared by morphometry, allozyme and cytogenetic analyses, as well as subjected to reciprocal crosses, in order to clarify the taxonomic status of the latter. No differences were found to distinguish the two species and the crosses between them yielded progeny. T. infestans populations presented four haplotypes that could be separated in two clusters: one formed by the samples from Bolivia (Andes and Chaco) and the other formed by samples from Argentina and Brazil. Silvatic and domestic T. infestans populations from Bolivia (Andes) were genetically identical. PMID- 10677725 TI - Trypanosoma cruzi interaction with its vectors and vertebrate hosts. PMID- 10677726 TI - Immunopathology of Chagas disease - a historical overview. PMID- 10677727 TI - Integration of Trypanosoma cruzi kDNA minicircle sequence in the host genome may be associated with autoimmune serum factors in Chagas disease patients. AB - Integration of kDNA sequences within the genome of the host cell shown by PCR amplification with primers to the conserved Trypanosoma cruzi kDNA minicircle sequence was confirmed by Southern hybridization with specific probes. The cells containing the integrated kDNA sequences were then perpetuated as transfected macrophage subclonal lines. The kDNA transfected macrophages expressed membrane antigens that were recognized by antibodies in a panel of sera from ten patients with chronic Chagas disease. These antigens barely expressed in the membrane of uninfected, control macrophage clonal lines were recognized neither by factors in the control, non-chagasic subjects nor in the chagasic sera. This finding suggests the presence of an autoimmune antibody in the chagasic sera that recognizes auto-antigens in the membrane of T. cruzi kDNA transfected macrophage subclonal lines. PMID- 10677728 TI - The role of the immune response on the development of severe clinical forms of human Chagas disease. PMID- 10677729 TI - Immunopathology of cardiomyopathy in the experimental Chagas disease. AB - The mechanisms by which Trypanosoma cruzi causes cardiomyopathy and induces neuronal destruction are discussed in this paper. The results suggest that autoimmunity in the chronic phase is the main cause of the progressive cardiac destruction, and that autoreactivity is restricted to the CD4+ T cell compartment. During the acute phase, the neuronal and cardiac fiber destruction occurs when ruptured parasite nests release T. cruzi antigens that bind to the cell surface in the vicinity which become targets for the cellular and humoral immune response against T. cruzi. The various factors involved in the genesis of autoimmunity in chronic T. cruzi infection include molecular mimicry, presentation of self-antigens and imbalance of immune regulation. PMID- 10677730 TI - Human chronic chagasic cardiopathy: participation of parasite antigens, subsets of lymphocytes, cytokines and microvascular abnormalities. AB - This article tries to demonstrate by new pathological findings (with the use of immunohistochemical technique and confocal laser microscopy) that chronic chagasic cardiomyopathy is a result of multiple factors involving myocarditis, immunodepression, severe fibrosis and microvessels dilatation and that all of these alterations are probably directly related with the presence of Trypanosoma cruzi parasites in the host associated with inadequate immunological response of the host. PMID- 10677731 TI - Mouse as a model for Chagas disease: does mouse represent a good model for Chagas disease? PMID- 10677732 TI - Immunosuppressive drugs as a tool to explore immunopathology in experimental Chagas disease. PMID- 10677733 TI - Evolution on the immunopathology of Chagas disease. PMID- 10677734 TI - The role of tissue-infiltrating T cells in immunopathology of Chagas disease. PMID- 10677735 TI - Development in the etiologic diagnosis of Chagas disease. PMID- 10677736 TI - Evolution of knowledge on the etiological diagnosis of chagasic infection. PMID- 10677737 TI - Serological diagnosis of Chagas disease with purified and defined Trypanosoma cruzi antigens. PMID- 10677738 TI - Xenodiagnosis. PMID- 10677739 TI - Changes in the hemoculture methodology improve the test positivity. PMID- 10677740 TI - Chagas disease diagnosis using polymerase chain reaction, hemoculture and serologic methods. PMID- 10677741 TI - Diagnosis of Trypanosoma cruzi chronic infections in humans: usefulness of the complement regulatory protein antigens and lytic antibodies in the control of cure. PMID- 10677742 TI - Polymerase chain reaction detection: new insights into the diagnosis of chronic Chagas disease. PMID- 10677743 TI - Screening and confirmation in chagas disease serology - a contribution. PMID- 10677744 TI - Clinical evolution and morbi-mortality in Chagas disease. PMID- 10677745 TI - Indeterminate form of Chagas disease. PMID- 10677746 TI - Risk of death due to chronic chagasic cardiopathy. AB - In this longitudinal study 5,710 people were included. The inclusion criteria were two positive serological results for Trypanosoma cruzi infection, 15 and 50 years old and no other demonstrable disease at the time of study. In the five year follow up 1,117 patients were lost. The follow up involved yearly evaluation of serology, clinical examination, X-ray of thorax, and ECG, for 4,593 patients and 263 were contacted at home because they did not assist for their clinical consultant. Time average of follow up was 5.3 years. Eighty nine (1.5%) of the 4,593 patients died during the follow-up period, 63 (71%) by cardiac insufficiency (CI) and 26 (29%) by severe ventricular arrhythmias. Diagnosis of cardiomegaly was present in all the patients with diagnosis of CI and in 15 (5%) of the patients with diagnosis of arrhythmias. The ECG alterations of these patients show 61 right bundle branch block (RBBB), associated or not with left anterior hemiblock (LAHB), 47 pathological Q wave and 70 primary repolarization alterations; 61 had polyfocal ventricular arrhythmia. The death rate was similar in the sexes and was more frequent between 40 and 50 years of age. Information on 1,380 recuperated patients shows that 15 died with no previous symptoms and without medical assistance and were interpreted as sudden death. The latest ECG in three follow-up of these patients indicates (before death) that only one had normal study and 14 presented 12 RBBB; 9 LAHB; 7 isolated ventricular arrhythmia; 10 repolarize alterations; 2 pathological Q wave, 10 patients of them with RBBB and repolarize alterations. In all the cases we had people between 35 and 43 years old, 9 men and 6 women. This study shows that in Chagas disease is possible to differentiate two risk groups. A low risk death group that have normal ECG and clinical evaluation during the follow up, and a high risk group associate ECG with RBBB and primary alterations of repolarization and/or inactivation zones with not annual clinical evaluation. PMID- 10677747 TI - Sudden death in patients with Chagas disease. PMID- 10677748 TI - Chagas disease and immunosuppression. PMID- 10677749 TI - Longitudinal radiological study of the esophagus in Chagas disease. PMID- 10677750 TI - Criteria of Chagas disease cure. PMID- 10677751 TI - Etiological treatment for infection by Trypanosoma cruzi. PMID- 10677752 TI - Clinical treatment of the digestive form of Chagas disease. PMID- 10677753 TI - Chagas disease surgery. PMID- 10677754 TI - Parasitological cure of Chagas disease: is it possible? Is it relevant? PMID- 10677755 TI - Chagas disease: criteria of cure and prognosis. PMID- 10677756 TI - Treatment of Trypanosoma cruzi infection in the undetermined phase. Experience and current guidelines of treatment in Argentina. PMID- 10677757 TI - Climatic factors related to Chagas disease transmission. PMID- 10677758 TI - Genetic variability in Brazilian triatomines and the risk of domiciliation. PMID- 10677759 TI - The process of domestication in Triatominae. PMID- 10677760 TI - Chagas disease: from bush to huts and houses. Is it the case of the Brazilian Amazon? AB - Two of the major problems facing the Amazon - human migration from the other areas and uncontrolled deforestation - constitute the greatest risk for the establishment of endemic Chagas disease in this part of Brazil. At least 18 species of triatomines had been found in the Brazilian Amazon, 10 of them infected with Trypanosoma cruzi, associated with numerous wild reservoirs. With wide-range deforestation, wild animals will perforce be driven into other areas, with tendency for triatomines to become adapted to alternative food sources in peri and intradomicilies. Serological surveys and cross-sectional studies for Chagas disease, carried out in rural areas of the Rio Negro, in the Brazilian Amazon, showed a high level of seropositivity for T. cruzi antibodies. A strong correlation of seroreactivity with the contact of gatherers of piacava fibers with wild triatomines could be evidenced. PMID- 10677761 TI - Epidemiology and dynamics of the vectorial transmission of Chagas disease. PMID- 10677762 TI - Epidemiology of Chagas disease in Ecuador. A brief review. AB - Chagas disease is a complex public health problem that has been underestimated in Ecuador. Here we review the relevant published information, and present unpublished and new data that help to understand the current Chagas disease epidemiological situation and its evolution in the country. Three main characteristics have been identified: (i) persistence of Trypanosoma cruzi transmission in already known foci; (ii) a marked endemicity in some urban areas of Guayaquil; and (iii) the transformation of new Amazon foci into truly endemic areas. The situation in other suspect areas remains uncertain. Five Triatominae species have been implicated in the transmission of T. cruzi to people in Ecuador (Triatoma dimidiata, Rhodnius ecuadoriensis, R. pictipes, R. robustus and Panstrongylus geniculatus), but some others may also play a role in some areas (P. rufotuberculatus, P. howardi, T. carrioni and P. chinai). Other Triatominae reported seem to have little or no epidemiological relevance (T. venosa, T. dispar, Eratyrus mucronatus, E. cuspidatus, P. lignarius and Cavernicola pilosa). High frequency of acute cases and severe chronic disease has been observed. Although cardiomyopathy is more frequent, serious digestive disease is also present. It is estimated that around 120,000-200,000 people may be infected. 2.2 to 3.8 million people are estimated to live under transmission risk conditions. PMID- 10677763 TI - Considerations on the epidemiology and transmission of Chagas disease in the Brazilian Amazon. PMID- 10677764 TI - Potential for domestication of Panstrongylus geniculatus (Latreille, 1811) (Liemiptera, reduviidae, triatominae) in the municipality of Muana, Marajo island, state of Para, Brazil. PMID- 10677765 TI - Progress towards interruption of transmission of Chagas disease. PMID- 10677766 TI - Elimination of vector-borne transmission of Chagas disease. AB - The control of the vector-borne transmission of Chagas disease in Brazil was organized as a national program in 1975, when two large entomological and sero epidemiological surveys were conducted in the country in order to identify areas at highest risk of transmission and to guide interventions regarding the chemical treatment of domestic vectors of the disease. The authors present the baseline data gathered through these studies and compare them with more recent data. The evaluation performed shows that the transmission by Triatoma infestans is virtually interrupted and that the transmission by other native species of triatominae from different regions of the country is possibly very low. It is emphasized the need to maintain permanent actions of entomological surveillance in order to prevent recurrent transmission. PMID- 10677767 TI - Interruption of Chagas disease transmission in the Andean countries: Colombia. PMID- 10677768 TI - Elimination of the vectorial transmission of Chagas disease in Central American countries: Honduras. PMID- 10677769 TI - Chagas infection transmission control: situation of transfusional transmission in Brazil and other countries of Latin America. AB - The transmission of the transfusion-associated Chagas disease is an important mechanism of its dissemination in several Latin American countries. The transmission risk depends on five factors: prevalence of infection in blood donors, degree of serological coverage, sensibility of used tests, safety of obtained results and infection risk. The Southern Cone Iniciative set off by the Pan-American Health Organization, in 1991, is contributing to the implementation of blood law in each endemic country, and to reduce the risk of transfusional transmission of this horrible disease. Despite the clear improvement of Brasilian hemotherapy after 1980 (with the creation of the Blood National Program - Pro Sangue) and the significant reduction of the chagasic infection among its blood donors; socio-economic, politic and cultural unlevels, prevent it from reaching the necessary universality and security. In order to assure both, the Brazilian Ministry of Health decided to restructure its blood system. In May, 1998, a great program was launched, to reach a specific goal: Blood - 100% with quality safety in all its process until 2003. It was divided in 12 projects, intends to guarantee the quality and self sufficiency in blood and hemoderivates. PMID- 10677771 TI - Recommendations from a satellite meeting. PMID- 10677770 TI - Differences of susceptibility of five triatomine species to pyrethroid insecticides - implications for Chagas disease vector control. AB - As pyrethroids are presently the favored group of insecticides to control triatomines, we performed a series of bioassays to determine the intrinsic activity of some of the main compounds used in the control campaigns, against five of the main species of triatomines to be controlled. Comparing the insecticides it can be seen that lambdacyhalothrin is more effective than the other three pyrethroids, both considering the LD50 and 99 for all the three species with comparable results. On Triatoma infestans the LD50 of lambdacyhalothrin was followed by that of alfacypermethrin, cyfluthrin and deltamethrin. On Rhodnius prolixus the sequence, in decreasing order of activity, was lambdacyhalothrin, alfacypermethrin, deltamethrin and cyfluthrin. Some modifications can be seen when we compare the LD99, that has more to see to what happens in the field. T. brasiliensis showed to be as sensible to lambdacyhalothrin as T. infestans, the most susceptible for this product. By the other side T. sordida is the least susceptible considering the LD99 of this insecticide. PMID- 10677772 TI - The Mount Sinai Hospital--a brief history. AB - In 1852, The Jews Hospital was founded for the increasing number of Jews in New York. It opened in 1855 with 45 beds on West 28th Street; 92% of the patients were indigent. In 1864, the hospital formally became nonsectarian and, in 1866, changed its name to The Mount Sinai Hospital. The medical staff was primarily Jewish, because until relatively recently, it was difficult for Jewish doctors to obtain postgraduate training or specialist posts at major New York hospitals. As the Jewish population moved uptown, so did The Mount Sinai Hospital: in 1870 to 66th Street, and in 1904 to 100th Street, with 456 beds, growing with new buildings and services to the current 1100 beds, 50,000 discharges, 400,000 inpatient days and 300,000 outpatient visits each year. Services increasingly became specialized, and then subspecialized. Key innovations included the choice of interns by competitive examination (1872), an advisory Medical Board (1872), the Nurse Training School (1881), the library (1883), the Alumni Association (1896), a professional medical hospital administrator (1903), research laboratories (1904), clinicopathological conferences (1905), the Social Services Department (1906), postgraduate teaching programs (1923), full-time chiefs of clinical services (1944), the dedication of the Mount Sinai School of Medicine (1968), and the merger in 1998 into the Mount Sinai-New York University Medical Center. PMID- 10677773 TI - Gastroenterology and hepatology as subspecialties. AB - Gastroenterology grew as a subspecialty in Germany in the 19th century. In the 1880s and 1890s, Austrian and German clinics were attended by American physicians who, on returning to the U.S., founded the American Gastroenterological Association in 1897. The creation of a subspecialty board, however, had to wait until 1941. At The Mount Sinai Hospital, Dr. A.A. Berg was appointed Surgeon in 1899. His practice focused on the alimentary tract, which in 1910 became one of the four surgical specialties. In 1914, further subdivision led to the stomach and duodenum becoming additional specialties. In 1917, wards were endowed for Dr. Berg's specialty. The first Mount Sinai physician to have an interest in gastroenterology was Morris Manges, but the first to limit his practice to gastroenterology was Dr. Edward Aronson, for whom a specialist outpatient division was formed in 1913. Aronson died in 1922 and was succeeded by Dr. Burrill Crohn, who was followed in 1934 by Dr. Asher Winkelstein; all three collaborated closely with the surgeons, physiologists and biochemists. In 1958, Dr. Henry Janowitz became chief of the Division of Gastroenterology; he was succeeded in 1983 by Dr. David Sachar, who was followed in 1999 by his associate Dr. Steven Itzkowitz. In 1958 Dr. Fenton Schaffner became chief of the Division of Hepatology (now headed by Dr. Paul Berk), and in 1979 Dr. LeLeiko became chief of Pediatric Gastroenterology. PMID- 10677774 TI - Morris Manges and Edmund Aronson. PMID- 10677775 TI - Burrill B. Crohn (1884-1983). PMID- 10677776 TI - Asher Winkelstein (1893-1972). PMID- 10677777 TI - Franklin Hollander (1899-1966). PMID- 10677778 TI - Gastroenterology and hepatology--the diagnostic data. AB - The Annual Reports of the Mount Sinai Hospital from the 1850s, and the Mount Sinai Hospital Reports for 1897-1906, make it possible to trace the discharges of gastroenterological inpatients, and (for a few years) of outpatients. Fully computerized diagnostic data have only been available since 1986. In the 19th century, about 20% of the outpatients had digestive disorders, the commonest of which were gastralgia/gastritis/dyspepsia, gastroenteritis, oropharyngeal complaints and constipation. A similar proportion of inpatients had digestive diagnoses, but the four disorders listed above decreased markedly in the second half of the 19th century, so that by the turn of the century the commonest diseases were typhlitis (appendicitis), hemorrhoids and other anal problems. By the 1990s, digestive diseases accounted for only 5% of total admissions, hepatobiliary diagnoses being the commonest group. Some cancers such as gastric and esophageal showed little change, while colorectal increased markedly. Some newly recognized diseases, such as peptic ulcer, waxed and then waned, while colitis and regional enteritis came and have continued to increase. Other new diagnoses, such as autointoxication and visceroptosis, flashed into prominence and then disappeared totally, presumably because they were nondiseases. PMID- 10677779 TI - The esophagus. AB - Original investigations and descriptions of the radiographic findings and techniques of evaluation of the esophagus and esophagogastric junction were made at The Mount Sinai Hospital by Drs. Bernard S. Wolf and colleagues in the third quarter of the 20th century. These included basic descriptions of peptic ulceration of the esophagus, the gastric lined esophagus, definitions of hiatus hernia, terminology of the esophagogastric junction, use of the barium pill and correlations of cineradiology with manometry. PMID- 10677780 TI - Gastric secretion. AB - The European gastric test meal was widely used in The Mount Sinai Hospital in the 1890s and early 1900s, but was then abandoned diagnostically after the introduction of gastroscopy and radiology. The fundamental methodological advances of Franklin Hollander led to his quantitative formulation of the ionic concentrations of the gastric acid parietal and nonparietal components, followed by his insulin test for completeness of vagotomy. PMID- 10677781 TI - Acid secretion after gastric operations. AB - In the early 20th century, the commonest surgical treatment of peptic ulcer was gastroenterostomy. Crohn and Wilensky demonstrated that this operation did not achieve its aim of markedly reducing gastric acidity or of accelerating motility. These results were highly controversial, but led to Lewisohn visiting Haberer in Austria in 1922, and convincing Dr. A.A. Berg to abandon gastroenterostomy and use partial gastrectomy as the standard ulcer operation, with additional vagotomy in those patients with duodenal ulcer with high acidity. In 1929, a few patients were treated by vagotomy and gastrojejunostomy by Dr. Ralph Colp, with discouraging results. It was only in the 1940s that Mount Sinai surgeons adopted transthoracic or subdiaphragmatic vagotomy and gastroenterostomy (or later, pyloroplasty) as their standard, effective acid-lowering treatment of peptic ulcers. PMID- 10677782 TI - The gastric mucosal barrier. AB - Most gastroduodenal ulcer disease results from a weakness in the normal gastric mucous barrier against the penetration of acid secreted by the stomach. Based on meticulous and insightful research, the distinguished physiologist Franklin Hollander hypothesized that the stomach is protected against its own acid secretion by a dynamic two-component mucus-mucosal barrier. Hollander and his co workers defined the physical and chemical characteristics of the mucus components of this barrier, as well as the defense provided by the surface epithelial cell layer, which he viewed as the second line of defense (the second component). Barrier investigators at Mount Sinai demonstrated the effects of impairment of barrier function with resultant increased back-diffusion of acid, and they defined the consequences of this acid penetration into the gastric epithelium. The contribution of these workers included important observations on the natural impermeability of the gastric corpus and fundus as well as the normally increased permeability of the antrum. They also presented evidence on the role of bile in duodenogastric reflux in gastric ulcer disease and the presence of impaired barrier function in patients with gastric ulcer and pernicious anemia. Further studies included demonstration that stress and carcinogens could disrupt the gastric mucosal barrier. Disruption of the barrier, in turn, was shown to allow carcinogenesis to occur by permitting the absorption of certain carcinogens which otherwise are warded off by the barrier. The Hollander two-component gastric mucosal barrier hypothesis has, in recent years, been increasingly validated by experimental data coming from other laboratories. PMID- 10677783 TI - Peptic ulcer. AB - Indigestion and heartburn have been described for thousands of years, but it was only in the 16th century that the disease peptic ulcer was established by autopsy. At first, only gastric ulcers were identified. In the 18th century, duodenal ulcers, most of which were fatal cases after perforation or hemorrhage, were seen. In the 19th century, when autopsy became a common, even routine, hospital procedure, uncomplicated acute and chronic ulcers were found and then correlated with symptoms. Thus, our current clinical understanding dates from the 1820s, by which time peptic ulcers were being reported in the U.S. It is unclear why gastric ulcers were not diagnosed at The Mount Sinai Hospital until 1873 and duodenal ulcers until 1885. However, after that time both conditions were diagnosed frequently, and they rapidly became common and were treated medically and surgically. PMID- 10677784 TI - Treatments of peptic ulcer. AB - From the late 19th century, Mount Sinai gastroenterologists declared their scepticism of the efficacy of all recommended treatments of peptic ulcer, and looked forward to trials which could distinguish between sequence and consequence, between association and causation. The rationale of all the early studies was to reduce gastric acidity, but it soon became clear that any neutralization by single doses of antacids was brief and ineffective. Winkelstein s demonstration that patients with duodenal ulcer had higher acidities not only before and after meals but also through the night hours led him to introduce a new treatment, the alkalinized intragastric milk drip together with atropine. One of the earliest controlled clinical trials at Mount Sinai compared different antacid regimes and showed that pH values above 3.5 were achieved in only about half of the patients on the various drips. When the new anticholinergic drugs were developed in the 1950s, they were found to produce sustained hypoacidity and were tried as maintenance treatment, as an alternative to acid-lowering operations. The third Mount Sinai approach was to attack the machinery of the acid-producing cell itself by an inhibitor of the enzyme producing hydrogen ions. In 1939, this enzyme had been thought to be carbonic anhydrase, but when Janowitz and Hollander tested its inhibitor, acetazolamide, and showed marked but very brief acid inhibition, they concluded that its action was too brief to be therapeutically useful. The problem was to be solved decades later by H2 receptor blockers from Britain and H+K+ATPase inhibitors from Sweden. PMID- 10677785 TI - The pancreas. AB - Pancreatic secretion was first studied at The Mount Sinai Hospital by Crohn in 1912, but measurements of pancreatic enzymes in duodenal aspirate or feces were found unhelpful in diagnosis. Such pancreatic tests fell into disuse because of advances in radiology of the biliary tree in the 1920s. Once extracts of secretin and cholecystokinin-pancreozymin became available from Sweden in the 1930s, it became possible for the biochemist Franklin Hollander and the surgeon David Dreiling to develop pancreatic secretion tests into practical procedures for the diagnosis of benign and malignant diseases of the pancreas and biliary tree, and produce physiological studies of the mechanisms of ion transport. With more purified hormones, it became possible to measure maximum (alkaline) bicarbonate output of the pancreas analogous to the maximal acid response of the stomach to an augmented histamine test, and to determine whether patients with duodenal ulcer had decreased neutralization of gastric acid in the duodenum. Clinical studies were also directed to the pathophysiology of acute relapsing and chronic pancreatitis and carcinoma. However, advances in imaging and endoscopy have now shifted the thrust of pancreatology. PMID- 10677786 TI - The history of liver disease at The Mount Sinai Hospital. AB - Diseases of the liver and biliary tract interested the physicians of The Mount Sinai Hospital from the time the hospital started until the present. Indeed, the institution has become a well-recognized center for the study of the liver and its diseases. During the first 75 years of the hospital, there were many admissions for hepatobiliary diseases, resulting in many case reports. The evolution of the hospital into a teaching hospital brought with it a more systematic method of studying diseases, not only in Pathology under Paul Klemperer, but in clinical chemistry and microbiology as well. Liver biopsy was also attempted. With the arrival of Hans Popper in 1957, the emphasis shifted to coordinated studies of structure and function under normal circumstances and in diseases as they progressed. Soon, Liver Diseases (Hepatology) were split from Gastroenterology, with Fenton Schaffner as the first chief. Over the next 30 years, more than 1000 papers, chapters and books were published. The main areas of research were fibrosis, cholestasis (especially morphology and bile salt metabolism), toxic liver injury, metabolic transformations and carcinogenesis. Primary biliary cirrhosis and viral hepatitis were and continue to be special interests. Fellows from all over the world were trained and many moved on to leadership positions. Although he was active in the development of the liver transplant program, Popper did not live to see its start. A new generation of hepatologists maintains the interest and position of The Mount Sinai Hospital in this important field of medicine. PMID- 10677787 TI - Alcohol and the liver: metabolism of alcohol and its role in hepatic and extrahepatic diseases. AB - Dr. Charles S. Lieber conducted clinical and experimental studies for more than four decades (three at Mount Sinai and the Bronx VA Medical Centers) with emphasis on liver, nutrition and GI pathophysiology. His major contributions include elucidation of the pathogenesis of alcoholic liver disease, by demonstrating the toxic role of alcohol and describing associated metabolic disorders. This was achieved through judicious clinical studies and newly developed rodent and primate models with the administration of ethanol in liquid diets. The mechanisms of various pathological and metabolic effects of ethanol were clarified, including hyperlipemia (with the rise in HDL), hyperuricemia, the role of acetaldehyde toxicity and alcohol-induced oxidative stress. The latter, including glutathione depletion, was corrected by S-adenosyl-1-methionine given to alcohol-fed baboons; the compound is now being used successfully for the treatment of patients with alcoholic liver disease in Europe. Alcoholic cirrhosis was produced for the first time in nonhuman primates and shown to be fully prevented by polyenylphosphatidylcholine, which is now being tested in a multicenter clinical trial. Lieber also discovered a new (microsomal) pathway of ethanol metabolism, responsible for the tolerance to ethanol and for several clinically important toxic interactions with other drugs (e.g., acetaminophen), anesthetics, industrial solvents, carcinogens, as well as retinol and b-carotene, with narrowing of their therapeutic window. His work defined the role of the stomach in ethanol metabolism, description of corresponding gender differences, cloning (for the first time) of the gene for sigma ADH (a newly-recognized gastric alcohol dehydrogenase isozyme) with its chromosomal localization, and the discovery of the effects of commonly used medications (e.g., H2 blockers and aspirin) on the activities of the enzyme and on blood alcohol levels in social drinkers. Lieber was among the first to use antibiotics for the elimination of gastric bacterial urease and its ammonia production in man, thereby alleviating chronic gastritis and hypoacidity, with attenuation of hepatic encephalopathy in cirrhotics. He promoted early detection and treatment of heavy drinkers before their social or medical disintegration, by defining precirrhotic lesions and markers of alcohol consumption. CONCLUSIONS: The research of Dr. Lieber and his group yielded a better understanding of the pathogenesis of common hepatic, gastric and nutritional disorders, with elucidation and prevention of serious toxic alcohol-drug interactions and the development of methods for early recognition and more effective approaches to prevent and treat liver and gastrointestinal diseases. PMID- 10677788 TI - The role of clinical risk factors in the prediction of future risk. PMID- 10677789 TI - Predicting a second hip fracture. AB - In an attempt to identify a high-risk cohort of patients, who could be offered preventive therapy, we assessed patients who had suffered one hip fracture. A total of 394 patients were prospectively followed to determine those who had suffered a second fracture. Entry bone mass of the unfractured hip and total body was examined by dual X-ray absorptiometry (DXA) and of the os calcis, by quantitative ultrasound (QUS), along with various clinical parameters. The relative risks in the QUS parameters did not reach significance, except for broadband ultrasound attentuation as measured by the McCue CUBA Clinical, whereas femoral neck and total body bone mineral density also reached significance. Lowest quartile body weight was also a significant risk factor as were occurrence of a new fall and poor mobility score. Using Receiver Operator Characteristic curves, we found no significant differences between DXA trochanter or for the Mini Mental State Examination score in predicting those who sustained a second hip fracture. In this elderly group risk factors are almost as good as bone mass at predicting those who will sustain a second hip fracture. Low body weight and poor mobility could be used as triggers for the use of preventive therapy without the use of bone mass measurements and to target expensive preventive therapy to reduce fracture risk. PMID- 10677790 TI - Influence of strontium on bone mineral density and bone mineral content measurements by dual X-ray absorptiometry. AB - The presence of Sr in bone influences bone mineral density (BMD) and content (BMC) measurements by dual-energy X-ray absorptiometry (DXA). This interaction is of interest, since strontium ranelate (S12911) demonstrated positive effects on bone metabolism in various animal models of osteoporosis, and is currently being evaluated for treatment of postmenopausal osteoporosis. The present in vitro study aimed to determine adjustment factors for DXA measurements of BMC and BMD at different Sr concentrations in order to estimate the corresponding values that would have been measured without Sr. A series of mixtures of Ca and Sr hydroxyapatites were prepared, with biologically relevant Sr/Ca ratios ranging from 0 to 3.5 mol/mol%, and a constant total concentration of divalent cations (145 mmol). The mixtures were conditioned in plastic dishes 4.5 cm in diameter, to obtain an areal density close to the human vertebral mineral density of 0.7 1.1 g/cm(2). DXA measurements of the mixtures were made with a wide range of different instruments and various acquisition modes. A direct linear relationship (r(2) > 0.99) was found between strontium content and overestimation of BMD and BMC. There were no significant differences in adjustment factors for BMC or BMD between the different machines or acquisition modes, and the presence of Sr in the water bath used to mimic soft tissues did not affect the accuracy and precision of the method. This demonstrates that reliable DXA determinations of BMD may be carried out in the presence of Sr, and may be interpreted in terms of calcium hydroxyapatite equivalent if the bone Sr content of the measured bone is known. The same adjustment factor (10% overestimation for 1 mol/mol% Sr) can be used for all presently available types of instrument and acquisition modes. PMID- 10677791 TI - Measurement of distal forearm bone mineral density: can different forearm segments be used interchangeably ? AB - Many different forearm sites have been used for the measurement of bone mineral density (BMD) and prediction of risk of future fracture among community dwelling populations. In populations where bone densitometry of peripheral sites may be the most cost effective and practical means of measuring BMD, such as the nursing home population, knowing the characteristics of forearm BMD measures would be beneficial. The purpose of this study was to assess the relationship of four common commercially available measures to each other and to estimate the inter- and intrarater reliability of the measures in a sample of nursing home residents as a first step toward identifying appropriate forearm measurement sites. These sites were the distal radius, the distal ulna, a composite of the distal radius and distal ulna, and the ultra distal radius. BMD measurements on 48 nursing home patients were obtained using single X-ray absorptiometry. Inter- and intrarater reliability was excellent at all four sites (interclass correlation coefficients > 0. 85). Moderate to high correlations (0.84-0.91) between the distal radius and ultra distal radius sites of the forearm suggest that these measures may be interchangeable. Although not directly assessed here, differences in bone composition among forearm sites may partially explain moderate rather than high correlations between sites and may affect the ability of each site to predict future fractures. Thus, different forearm sites may be used interchangeably for diagnostic purposes; however, the prognostic value of each site remains unknown. PMID- 10677792 TI - Quantitative ultrasound in the assessment of skeletal status in uremic patients. AB - Renal osteodystrophy (ROD) can be characterized by both high (HT) and low (LT) bone turnover states. Although bone biopsy remains the "gold standard" to diagnose ROD, noninvasive tools for the diagnosis and follow-up of such bone disease are desirable. Recently, ultrasound (US) techniques, proposed to assess skeletal status, have been shown to be correlated not only with bone density but also with bone quality. We have investigated 98 patients on chronic hemodyalisis (HD) and 98 healthy, sex- and age-matched subjects. Amplitude-dependent speed of sound (AD-SOS) and ultrasound bone profile score (UBPS) at phalanxes and speed of sound (SOS), broadband ultrasound attenuation (BUA), and a quantitative ultrasound index (QUI/stiffness) at the heel were performed in both groups. In all subjects intact parathyroid hormone (PTH), total alkaline phosphatase (T ALP), bone isoenzyme alkaline phosphatase (B-ALP), and carboxy-terminal telopeptide of type I collagen (ICTP) were assessed. All US parameters were significantly lower in the hemodialysis group than in control subjects. Moreover, among US parameters only AD-SOS and UBPS showed a significant correlation with PTH, T-ALP, and B-ALP. Dialytic age showed a modest, but significant correlation only with US parameters at the phalanxes. On the basis of bone biochemical markers, we considered a group with high and a group with normal to low bone turnover. AD-SOS and UBPS, but not SOS, BUA, and stiffness were significantly (p < 0.01) lower in the high bone turnover than in low bone turnover group. Furthermore, in the high bone turnover group, parameters of the US phalanxes strongly correlated with B-ALP. Our results seem to demonstrate that US parameters are a useful tool in the assessment of skeletal status in patients on maintenance dialysis. PMID- 10677793 TI - Radiation dose to the patient and operator from a peripheral dual X-ray absorptiometry system. AB - Although peripheral dual X-ray absorptiometry (pDXA) scanners for measuring bone mineral density (BMD) in the forearm are known to produce an exceptionally low radiation dose to the patient, quantitative assessment of patient dose from pDXA procedures is important for reassuring patients about their safety. We have estimated the effective dose of radiation (ICRP-60) to the patient and also the scattered dose to the operator from a forearm BMD examination performed on a DTX 200 pDXA system (Osteometer Meditech, Hoersholm, Denmark). Measurements were performed using thermoluminescent dosimeters (TLD's) attached to the forearm phantom supplied by the manufacturer. The effective dose to a patient was estimated to be 0.1 microSv. At a distance of 1 m from the center of the forearm, the time-averaged scattered dose to the operator assuming scanning five patients per hour was measured to be <0.1 microSv/h. The dose rate over the outside surface of the DTX-200 in line with the primary X-ray beam was measured to be 1.4 microSv/h. These figures compare with a natural background radiation in the United Kingdom of 7 microSv/d. In conclusion the radiation doses from forearm pDXA to both patients and operator were found to be truly trivial. PMID- 10677794 TI - A comparison of calcaneus ultrasound and dual X-ray absorptiometry in healthy North American youths and young adults. AB - Quantitative ultrasound is the newest noninvasive method to be accepted for assessing bone mineral in adults. Heel ultrasound measurements correlate with bone density measurements by dual X-ray absorptiometry (DXA) and predict fracture risk in adults. Far less is known about the value of calcaneus ultrasound (CUS) in children. We determine spine, femoral neck, and whole-body bone mineral by DXA and heel bone mass by CUS in 125 youths (69 females, 56 males) ages 9-25 yr. CUS and DXA measurements of bone mass increased with age and pubertal development during adolescence in a parallel fashion. Among females, Tanner stage was a stronger predictor than age for all CUS and DXA measurements, and among males, pubertal stage was a stronger predictor for spine bone mineral apparent density (BMAD) and femoral bone mineral density (BMD). CUS measurements correlated moderately well with DXA measurements of the spine, femoral neck, and whole-body BMD and spine BMAD (r = 0.23-0.58, p < 0. 008). CUS warrants further study as a tool for assessing bone mineral acquisition in children. PMID- 10677795 TI - In vivo detection of structural differences between dominant and nondominant radii using peripheral quantitative computed tomography. AB - This cross-sectional study identifies differences in distal radial trabecular bone structure related to habitual loading patterns in the upper extremities using high-resolution peripheral quantitative computed tomography. As well, it determines whether measurements of these indices in one limb serve as satisfactory surrogates for the contralateral limb. The dominant and nondominant forearms of 106 adult volunteers (mean age [SD], 44.3[17.5] yr) were scanned and indices of trabecular bone structure (connectivity index [CI], maximum hole size [H(M)], and mean hole size [H(A)]) were determined at the distal radius. The images were also analyzed to determine bone density. For all subjects, H(M) is significantly smaller in the dominant radius (p < 0.01). Right-handed subjects (n = 96) have greater CI (p < 0.05) and smaller H(M) (p < 0.01) in the dominant radius. For the total group, the dominant limb has a greater mass (total and cortical bone mineral content, p < 0.01 and p < 0.05, respectively) and greater total bone volumetric density (p < 0.05). There are no significant differences between limbs for the group of left-handed subjects &lapr;n = 10). As expected, significant associations exist between side-to-side measurements of bone structure and density (p < 0.001). The correlation coefficients for connectivity index, H(M), and H(A) are 0.86, 0.85, and 0.87, respectively. For bone density, the between-limb associations are 0. 90, 0.73, and 0.92 for the total, cortical, and trabecular bone compartments at the distal radius. Differences in the structure of the trabecular bone network suggest that differential loading of the dominant limb preserves bone strength. PMID- 10677796 TI - Perspectives on bone mechanical properties and adaptive response to mechanical challenge. AB - The bones of the human skeleton serve a mechanical function besides providing a reservoir for calcium and hematopoietic homeostatis. When mechanically challenged, they usually respond and adapt; failure to do so can result in fracture. The mechanical behavior of bone is determined by bone mass and its material properties and by its geometry and architecture. Therefore, in vivo noninvasive measurements of bone mass, geometry, and structure can predict bone strength and are usually employed as a useful-if not always reliable-way to estimate bone fragility, whereas direct bone biomechanical testing in vitro can provide detailed information about mechanical strength. Because bone strains are likely to be regulators of bone mass and strength, exercise protocols designed to counteract the effects of osteoporosis should load the target bone with repeated high peak forces and high strain rates or high impacts on a long-term basis. Such a protocol creates varied strain distributions throughout the bone structure, producing short, repeated strains on the bone in directions to which it is unaccustomed. Exercise in this manner can maintain and perhaps increase bone mass and improve mechanical properties and neuromuscular competency, reducing skeletal fragility and the predisposition to falls. PMID- 10677797 TI - A review of clinical trials of therapies for osteoporosis using fracture as an end point. AB - As the population ages, fragility fractures grow in importance as a public health problem. The principal goal of osteoporosis therapy is primary and secondary fracture prevention. A growing choice of therapies is now available for the treatment of osteoporosis. In this article, we review their efficacy using fracture prevention as an end point. The considerable heterogeneity among studies with regard to patient age, past fracture history, fracture site, and analytical methods precludes the possibility of performing a meaningful meta-analysis. Fracture outcomes have been reported in clinical trials with calcium supplementation, vitamin D supplementation, estrogen replacement therapy (ERT), calcitonin, etidronate, alendronate, sodium fluoride (NaF), parathyroid hormone (PTH), and raloxifene. Compelling evidence for fracture prevention has been provided for calcium and vitamin D supplementation and alendronate treatment. Evidence of fracture prevention exists for ERT, raloxifene, calcitonin, etidronate, and PTH. Data on NaF are inconsistent. Across agents, there is a trend toward greater efficacy for patients at greatest risk of fracture. PMID- 10677798 TI - A marked increase in bone mineral density in a patient with glucocorticoid induced osteoporosis. AB - During the densitometric follow-up of a 73-yr-old male suffering from bullous pemphigoid and glucocorticoid-induced osteoporosis, a marked increase in bone mineral density (BMD) led to the diagnosis of an asymptomatic prostatic cancer with secondary diffuse bone lesions. This eventuality must be considered in patients with a rapid and unexplained gain in BMD. PMID- 10677799 TI - Perspective on assessment of vitamin D nutrition. AB - Although routine fortification of milk and a few other dairy products has been successful in preventing rickets in children, its impact on preventing vitamin D depletion in adults is less than satisfactory. The prevalence of vitamin D depletion in the elderly is on the rise again and appears to be more common than is currently appreciated. Several groups of individuals are at risk of developing vitamin D depletion, and a significant minority of otherwise healthy individuals is vitamin D insufficient. Unrecognized vitamin D depletion leads to secondary hyperparathyroidism, accelerates cortical bone loss, and increases the risk of hip fractures. With the availability of techniques to assess vitamin D nutrition, it is probably cost effective to routinely measure 25-hydroxyvitamin D levels in individuals at the greatest risk and in patients with various metabolic bone diseases to prevent vitamin D depletion. Early recognition and prompt treatment of vitamin D depletion improves functional well being of the individual, reduces morbidity related to bone loss and fractures, and is associated with a highly favorable cost/benefit ratio. PMID- 10677800 TI - PROQUAD: accreditation program of the Brazilian society for clinical densitometry. AB - In 1996, the Brazilian Society for Clinical Densitometry (SBDens) developed an accreditation program to be applied to all densitometry centers, under the supervision of the certified physicians from SBDens. This program is named Programa Nacional de Qualidade em Densitometria (PROQUAD) and was developed to verify at least three aspects of the bone densitometry practice: the functioning of equipment, the methods used by technicians in performing the scans, and how the physicians are analyzing the scans and interpreting the results. From the 360 certified physicians who are members of SBDens, nearly one-third are participating in this program. The final purpose of PROQUAD is to provide approved centers with a seal, to be stamped on their examinations. The seal will signify to referring physicians, and the community, which centers are working under high standards of quality. It has been 2 yr since PROQUAD was established, and the data obtained confirm the improvement of quality in the practice of densitometry in Brazil. Any input from other societies that could improve PROQUAD is welcomed. Ensuring good densitometry practices is a goal for which all densitometry societies must strive. PMID- 10677801 TI - Electroporation therapy of solid tumors. AB - The curative effects of some chemotherapeutic drugs are impeded by their poor permeation through the cell membrane. This limitation can be overcome by a novel approach called electroporation therapy (EPT), electrochemotherapy (ECT), or electrical impulse chemotherapy (EIC). The method involves application of brief electrical pulses, which destabilize the cell membrane barrier, allowing intracellular access of chemotherapeutic drugs that otherwise would not be able to penetrate the cell membrane effectively. EPT makes it possible to lower the drug dose, thereby relieving the patient of adverse side effects associated with conventional chemotherapy. Even with the lower drug dose, EPT has shown significantly higher efficacy than has conventional chemotherapy. The method is currently being evaluated clinically for treating various cancer indications using the anticancer drugs bleomycin or cisplatin. This article provides a historical perspective and current insights into this new modality of cancer treatment, including basic physical, biological, and medical facts about EPT; computer-assisted development of electrical pulse generators and electrodes necessary to create effective electrical fields in the treatment area; results of cancer cell and tumor treatments in vitro, in animals, and in humans; safety aspects of EPT; potential combined delivery of chemotherapeutic drugs and biological agents to reduce or eliminate metastatic disease; and intracellular delivery of DNA by electroporation for cancer gene therapy. PMID- 10677802 TI - Pharmacodynamic aspects of modes of drug administration for optimization of drug therapy. AB - Because of various pharmacodynamic properties, such as the nonlinearity of the concentration-effect relationship, activation of feedback homeostatic mechanisms, induction of pharmacodynamic tolerance, etc., administration of the same dose of drug by different modes is expected to produce different outcomes. This review clarifies the theoretical and practical aspects of the impact of different modes of drug administration on the magnitude of response, and hence on therapy outcomes. It discusses how the interrelationship between the pharmacodynamic properties and the drug input function affects the magnitude of response. To demonstrate this special dimension of drug therapy, relevant pharmacodynamic data was obtained for drugs with different therapeutic applications, including antibiotics, analgesics, diuretics, anti-cancer, anti-ulcer, anti-inflammatory, anti-hypertensive, lipid-lowering anti-parkinsonian, and immunosuppressive drugs. These examples provide guidelines for implementing the role of the mode of drug administration (including rate, schedule, and route of drug treatment) during drug development or optimization of drug therapy. PMID- 10677803 TI - Analysis of mean disintegration time and mean dissolution time by moment analysis using microcalorimetric curves. AB - The mean disintegration time (MDGT; mean time required for disintegration of tablets) and mean dissolution time (MDST; mean time required for drug dissolution) of water-soluble drugs from solid dosage forms were determined by moment analysis using microcalorimetric curves. Microcalorimetric curves for heat of dilution and for heat of dissolution of the drug were prepared, and the zeroth and first moments of the calorimetric curves were then calculated. The difference between the first moments of the curves for powder dissolution and tablet dissolution was taken to be the MDGT. The difference between the first moment of the curve for heat of dilution and that of the curve for heat of dissolution was taken to be the MDST. Nicotinic acid and D-mannitol were used as model drugs. The dissolution rate was determined by the conventional beaker method and also by the deconvolution method. The dissolution process could be traced well by moment analysis, as well as by the other methods employed. Moment analysis has some advantages: (a) both the MDGT and the MDST can be determined simultaneously; (b) it is applicable to many drugs that are soluble with heat evolution without the need for quantitative analysis of the drug. PMID- 10677804 TI - The release of 5-fluorouracil from microspheres of poly(epsilon-caprolactone-co ethylene oxide). AB - The purpose of this study was to evaluate the in vitro release of 5-fluorouracil from microspheres prepared using a novel triblock copolymer of epsilon caprolactone and ethylene oxide as the encapsulating material. Microspheres of poly(epsilon-caprolactone-co-ethylene oxide) were prepared by employing the "hot melt" method of microencapsulation. Microspheres were sized using sieve analysis and scanning electron microscopy (SEM). Release studies were performed using a custom-made rotating paddle dissolution apparatus. Copolymer microspheres, fabricated by the hot melt method were shown by electron microscopy to have smooth, nonporous surfaces. Drug-loaded microspheres were found to have a broad distribution of sizes, which was thought to be a consequence of the wide range of crystal sizes of the encapsulated unmilled drug. Nonlinear release kinetics were observed from microspheres in the size fraction 75-250 microns, with a pronounced "burst release" associated with the presence of drug at the surface of the microspheres. A specific delineation of the drug release mechanism was not possible due to rapid gelation, swelling, and subsequent dissolution of the microspheres that occurred on hydration. This work describes the preparation of microspheres that swell rapidly and coalesce together on hydration, accompanied by rapid drug release and copolymer dissolution over a 2-hr period. PMID- 10677805 TI - The stability of theophylline tablets with a hydroxypropylcellulose matrix. AB - The behavior of 40:60 anhydrous theophylline/hydroxypropylcellulose (HPC) direct compression tablets obtained using a variety of hydroxypropylcelluloses with low or medium-high degrees of substitution (L-HPCs and HPCs, respectively) was determined immediately following their preparation and after storage for 6 months at 20 degrees C and a relative humidity (RH) of either 70.4% or 93.9%. The lower relative humidity did not bring about hydration of the active principle in any formulation, but the higher relative humidity totally hydrated the drug in all except one L-HPC formulation, in which hydration remained incomplete. Both relative humidities caused significant tablet swelling, with L-HPC formulations being more affected than HPC formulations. Drug release was slowed by hydration of the active principle, but accelerated with tablet swelling. The lower relative humidity caused significant alteration of drug release characteristics in only two L-HPC formulations, release from which was accelerated, while the higher relative humidities only failed to cause such alterations in two HPC formulations, with release from all except one of the others slowed (in the exceptional formulation, which exhibited incompletely hydrated theophylline and the greatest swelling of all, release was accelerated). PMID- 10677806 TI - Development of matrix-based theophylline sustained-release microtablets. AB - Microtablets containing high theophylline content (from 60% to 80%) based on a Eudragit RS PO matrix were produced on a rotary tablet press. The influence of the compaction pressure, the plasticizer content used for the granulation of theophylline particles, and the amount of theophylline on the drug release were investigated. The effects of surface area and the addition of magnesium stearate as a hydrophobic agent on the drug release were studied. The storage stabilities of the release rate at room temperature and at 50 degrees C were also determined. Dissolution profiles expressed as percentage of theophylline dissolved were obtained over 8 hr in 900 ml of purified water at 37 degrees C and 75 rpm. It was observed that the compaction pressure (from 200 MPa to 250 MPa) had no effect on the theophylline release. The use of triethyl citrate (TEC) as a plasticizer in the granulation of theophylline enhanced the physical properties of the microtablets. Theophylline content in the range 60% to 80% did not affect the drug release. The theophylline release obtained was a function of the quotient surface area/tablet weight and therefore was dependent on the tablet diameter. To reduce the dissolution rates, magnesium stearate was added in a concentration up to 50% of the matrix material. Tablets of this hydrophobic formulation fulfilled the requirements of USP 23 for theophylline sustained-release preparations. Storage at room temperature for 3 months and at 50 degrees C for 2 months showed no significant influence on the theophylline release. PMID- 10677807 TI - Bioavailability of itraconazole in rats and rabbits after administration of tablets containing solid dispersion particles. AB - A tablet dosage form containing solid dispersions of itraconazole (Asd tablets) was prepared by using the spray-drying and wet granulation methods. The dissolution rate of itraconazole from Asd tablets was fast, with more than 90% released within 10 min, compared to less than 20% for a marketed product, Sporanox capsules. The oral absorption of itraconazole from Asd tablets was determined in rats and rabbits and was compared with that for Sporanox capsules. In the rat, there was no difference between the Asd tablets and Sporanox capsules in the mean area under the curve (AUC) (3089.5 +/- 4332.8 ng.hr/ml and 3653.9 +/- 2348.9 ng.hr/ml, respectively) and Cmax (295.0 +/- 344.5 and 390.5 +/- 169.4 ng/ml, respectively). Also, in the rabbit, no difference was found between the two products in the mean AUC (AUMC; 19357.9 +/- 5117.5 ng.hr/ml and 23382.2 +/- 6236.5 ng.hr/ml, respectively) and Cmax (766.4 +/- 276.5 and 1127.5 +/- 577.9 ng/ml, respectively). Despite the rapid in vitro release characteristics of itraconazole from the Asd tablets, the in vivo absorption of itraconazole was comparable to that of Sporanox capsules, with no difference in Tmax in both animal species. Serum levels of the major active metabolite hydroxyitraconazole were also measured. Itraconazole was rapidly converted to hydroxyitraconazole in both rats and rabbits, but there were species-specific differences in their pharmacokinetics. It is concluded that, in addition to drug solubility and dissolution characteristics, other formulation factors such as the physical state of the drug and the granulation process, may also need to be considered in the prediction of the in vivo absorption of itraconazole based on in vitro data. PMID- 10677808 TI - The influence of hydrocolloid patch composition on the bioavailability of triamcinolone acetonide in humans. AB - Triamcinolone acetonide (TACA) is a corticosteroid; it is used in the systemic and topical treatment of a variety of inflammatory conditions, including eczema and psoriasis. Conventionally, for topical use, the drug is formulated in a cream or ointment. However, it has been observed in vitro that percutaneous penetration of corticosteroids can be influenced by hydrocolloid patches. Corticosteroids produce a pallor or blanching when applied to the skin that correlates with anti inflammatory activity; this property has been used extensively as a bioassay. The aim of this study therefore was to evaluate the occlusive properties of a range of hydrocolloid patches containing TACA on the drug's penetration in vivo using visual assessment and a graded multiple measurement. The in vivo hydration of these dermatological patches was also investigated. Statistical analysis of the weight gains of patches containing either NaCMC 39% or pectin 39% showed that there was a significant difference in the rates of hydration of the two types of patch (p < .005). An increase in application time of the hydrocolloid patches allowed more TACA to be released, which was illustrated by an increase in both the maximum percentage total possible score (%TPS) values and AUC, although changes in the hydrocolloid composition did not significantly alter the blanching response. All of the patches adhered well, were unobtrusive to the normal activity of the wearers, and showed great potential for the convenient, localized, prolonged delivery of drugs to the skin. PMID- 10677809 TI - The skin permeation mechanism of ketotifen: evaluation of permeation pathways and barrier components in the stratum corneum. AB - To evaluate the pathways and barrier components in the stratum corneum (SC) for the permeation of ketotifen, the effect of delipidization on the permeation and partition was examined under several donor pHs. Assuming that ionized ketotifen (KTH+) and un-ionized ketotifen (KT) contribute independently in both permeation and partition, the intrinsic permeability coefficients and SC/water partition coefficients of both species were estimated. Delipidization enlarged the permeability of KTH+ 100 times. This suggested that the lipid phase functions as the barrier against KTH+. KT has an intrinsic permeability 100 times larger than that of KTH+. Delipidization did not result in a significant change in permeability of KT. This suggested that the permeability of KT through the lipid phase is comparable to that through the aqueous phase in delipidized SC; that is, the lipid phase functions as a highly permeable pathway for KT. On the other hand, the permeability coefficient of KT through delipidized SC was 1/34 of that through the pure aqueous layer, which had a thickness equivalent to SC. Since this suggests that the permeability of KT through the proteinaceous phase is much lower than that through the aqueous phase, the proteinaceous phase can be assumed to function as a barrier against the permeation of KT. From these results, it is concluded that the predominant permeation pathway for KT is through the lipid phase. The SC/water partition coefficient of KT was cut in half by delipidization, but the value was still more than 100. These results show that the proteinaceous phase functions not only as the barrier, but also as the depot for KT. The knowledge obtained here will be useful for formulation design and for the selection of enhancers in a transdermal therapeutic system of ketotifen. PMID- 10677810 TI - Application of a sensorial response model to the design of an oral liquid pharmaceutical dosage form. AB - In this paper, we discuss the application of a compartmental model to study the sensorial response, in terms of taste intensity versus time, in an oral solution for pharmaceutical use. The numerical model was developed from sensorial response curves obtained by a panel of three trained individuals. Parameter identification was carried out by means of a least-squares procedure that obtained the linear coefficients in the model by solving an exact linear least-squares problem conditional on the values of the nonlinear parameters for each iteration. Thus, nonlinear estimation was done in terms of the first-order kinetic parameters only, and ill-conditioning of the Hessian matrix present in these models was solved. Results of modeling for a set of formulations were used to determine the effects of various ingredients (sweeteners and an essence) on a baseline unflavored formulation of acetaminophen in a mixture of cosolvents. The first moment of the area under the curve of taste intensity versus time was found to be the best global indicator of taste for the purpose of product design. It was found that a mixture of sweeteners and an essence was the most efficient way of masking the bitter taste of this active ingredient. PMID- 10677811 TI - Biodegradable progesterone microsphere delivery system for osteoporosis therapy. AB - The purpose of this study was to formulate and characterize a controlled-release biodegradable delivery system of progesterone for the treatment or prevention of osteoporosis. Microspheres of progesterone were formulated using copolymers of poly(glycolic acid-co-dl-lactic acid) (PGLA 50/50 and PGLA 15/85) and poly(L lactic acid) (L-PLA) of similar molecular weight by the emulsion solvent evaporation technique. The effects of process variables, such as volume fraction, polyvinyl alcohol (PVA) concentration, polymer composition, and stir speed during preparation, on the yield, encapsulation efficiency (EEF), particle size distribution, in vitro release profiles of progesterone, and surface morphology of progesterone microspheres were investigated. Increasing the volume fraction from 9% to 22% increased the EEF without significantly increasing the yield; however, the rate of progesterone release from the microspheres decreased. Increasing the PVA concentration from 1% to 5% had no significant influence on the EEF, but the rate of progesterone release from microspheres increased. Polymer composition had no significant effect on the EEF, but had a significant effect on the particle size distribution, surface morphology, and release rate of progesterone from the microspheres. Stir speed did not have a significant influence on the EEF; however, stir speed influenced particle size distribution and the rate of progesterone release from microspheres of the same sieve-size range. The results suggest that controlled release of progesterone is possible by varying the different process variables, and that PGLA 50/50 provided the slowest release of progesterone. This should provide a means of delivering progesterone for months for the treatment or prevention of osteoporosis in postmenopausal women. PMID- 10677812 TI - Scale-up of an oil/water cream containing 40% diethylene glycol monoethyl ether. AB - The purpose of this study was to scale up an oil/water (o/w) cream formulation containing 40% diethylene glycol monoethyl ether (DGME), developed via 300-g laboratory batches in a 2(5-2) fractional factorial design, to 7-kg batch sizes in a Brogli-10 homogenizer. The o/w cream was manufactured via a standard phase inversion process in the Brogli-10 homogenizer. Partitioning studies of DGME were conducted in test tubes at ambient temperature and after 24 hr at 70 degrees C in a convection oven. Phase height was measured by vernier calipers. Microscopy studies of excipients with and without treatment with water or a DGME/water mixture were conducted with a Nikon microscope after equilibration at 35 degrees C for 24 hr. During creation of the 7-kg pilot-scale batches, congealed material was observed between the sweep agitation blade and the discharge port, where the Brogli-10 homogenizer is not temperature jacketed. Factors that increased the amount of congealed material were higher temperatures during primary emulsification and longer cooling times. Partitioning studies revealed that DGME resides in the aqueous external phase of this formulation. Microscopy studies revealed that DGME in the external phase of this cream has a profound impact on the solubility of certain solid, waxy excipients (e.g., cetyl alcohol and polyoxyethylene-2-stearyl ether) at 35 degrees C. From this study, it appears that DGME resides in the external phase of the o/w cream. During manufacturing, it is hypothesized that the presence of DGME in the external phase alters the solubility of certain solid, waxy excipients in the formula such that they no longer primarily reside in the internal oil phase. On cooling, these materials precipitate or congeal in the external phase. The fractional factorial experimental design at the 300-g laboratory scale did not predict the issues encountered during scale-up. Differences between laboratory scale and pilot plant scale that explain why this phenomenon was not seen during laboratory scale are differences in cooling times, nonjacketed or "cold spots" in the Brogli-10 homogenizer, and a low proportion of congealed material in relation to the total batch size (< 1.5%). PMID- 10677813 TI - The use of a hydrophobic matrix for the sustained release of a highly water soluble drug. AB - An experimental investigation into the use of a hydrophobic matrix to control the release of a highly water soluble drug was undertaken. Matrices consisting of hydrogenated vegetable oil and calcium sulfate with a 4% drug loading showed a sustained-release profile of up to 24 hr. The release mechanism from such matrices seemed to obey both root time kinetics and first-order behavior. Investigations showed that the effect of geometry had a significant effect on the drug release rate. PMID- 10677814 TI - Interaction of ubiquinone-10 with dipalmitoylphosphatidylcholine and their formation of small dispersed particles. AB - Stable aqueous dispersions of ubiquinone-10 (UQ) were obtained by cosonication with dipalmitoylphosphatidylcholine (DPPC) in the UQ mole fraction range 0.1-0.7. To clarify the dispersal mechanism, the dispersed particles were characterized, and the interaction between UQ and DPPC was investigated using several physicochemical techniques. Dynamic light scattering (DLS) measurements showed that the diameter of the dispersed particles was 50-70 nm. A limited amount of UQ was incorporated into DPPC bilayer membranes (approximately 5 mol%). The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein, and the volume in the UQ/DPPC particles decreased remarkably with the addition of UQ into small unilamellar vesicles of DPPC. The decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO4. These results indicate that the excess UQ separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer. PMID- 10677815 TI - An approach to controlled-release dosage form of propranolol hydrochloride. AB - It is possible to release a drug with only limited diffusion from a membrane coated system using osmotic pumping. In this study, a propranolol osmotic pump was produced by coating the core tablets with cellulose acetate. The effects of membrane thickness, pore size, and stirring rate on the release rate of propranolol hydrochloride were studied. It was found that the thickness of cellulose acetate membrane had a profound effect on the release rate of propranolol hydrochloride from the membrane-coated tablets. The results showed that, when the membrane thickness increased, the release rate of propranolol decreased. The drug release follows a zero-order release when the delivery orifice is between 200 and 800 microns, but when the delivery orifice size is increased to 1000 microns, the release kinetic is abnormal. Fluid dynamics have an important effect on the delivery rate of propranolol from this device; the delivery rate increases as a function of the fluid flow. The drug release is higher under a turbulent condition with high rate of stirring. PMID- 10677816 TI - Studies of the drug permeability and mechanical properties of free films prepared by cellulose acetate pseudolatex coating system. AB - Free films produced with cellulose acetate (CA) pseudolatex were prepared by the casting method. The effects of plasticizer concentration, drying temperature, and drying time on drug permeability and mechanical properties of free films were investigated by three-factor spherical second-order composite experimental design. The results were analyzed by the multivariable regression method. The experimental results indicated that plasticizer concentration, drying temperature, and drying time had complex effects on free film permeability and mechanical behavior. These results probably arise from the film-forming ability of CA pseudolatex particles at various conditions and the evaporation of plasticizer during the film-forming process. PMID- 10677817 TI - Pharmacokinetics and urinary excretion of DW116, a new fluoroquinolone antibacterial agent, in humans as a phase I study. AB - Pharmacokinetics and urinary excretion of the new fluoroquinolone antibacterial agent DW116 [1-(5-fluoro-2-pyridyl)-6-fluoro-7-(4-methyl-1-piperazynyl)-1, 4 dihydro-4-oxoquinolone-3-carboxylic acid, hydrochloride] following oral administration (200, 400, 600 mg) were studied in humans as a phase I study. The plasma concentration of DW116 declined monoexponentially with a half-life range of 16-22 hr. The area under the curve (AUC) and Cmax increased proportionally as the dose increased. The T1/2 and mean residence time (MRT) (28.3-30.9 hr) were independent of dose. The Tmax appeared within 3 hr (0.9-2.7 hr) after drug administration. The Ka ranged from 1.3 to 4.1 (hr-1). The plasma half-life was much longer, and Cmax was higher by about two- to three-fold than conventional fluoroquinolones. Urinary recovery of DW116 was 29.6-61.6% of the dose. The maximum excretion rate appeared within 4 hr and decreased continuously after drug administration. A urinary metabolite was not detected in the urine extract obtained before and after hydrolysis by beta-glucuronidase (from Escherichia coli); this was different from other fluoroquinolone antibacterial agents. Poor metabolism in the kidney may contribute to the good oral bioavailability, but due to the low recovery (< 60%) in urine, it is possible that DW116 is metabolized in the liver or other organs. PMID- 10677818 TI - Using online and offline change models to improve ICU access and revenues. AB - BACKGROUND: Hospital operational problems that span departments often present formidable challenges because they involve both processes and organizational relationships. Many improvement efforts fail because of relationship issues rather than a lack of understanding of system processes. Reflection on a recent change initiative led to the development of an integrated change model that includes both online and offline components. The online component draws on performance improvement models that provide concepts and tools for use in team meetings to improve processes. The offline component borrows from an earlier tradition of change management that offers guidelines for individuals or teams desiring to be change agents. METHODS: The integrated change model was applied in 1997 at The Johns Hopkins Hospital, Baltimore, to reduce ambulance bypass hours, a chronic problem resulting in $6.7 million in lost revenue annually. The goal was to reduce red alert hours per month by 50%. Three Plan-Do-Study-Act (PDSA) cycles were implemented to test change concepts. RESULTS: There was a significant reduction in red alert hours after the change initiative, with an estimated $6 million in additional hospital revenue. DISCUSSION: The integrated change model may serve as a prototype for improving complex problems in which improving organizational relationships may be as difficult as improving processes and is likely to require a significant amount of work offline. For example, this approach may be particularly helpful for improving processes that span departments or functional units such as reducing cycle times for admissions, first-dose medications, as well as in building and improving integrated delivery systems. The model awaits further testing and evolution. PMID- 10677819 TI - Improving management of atrial fibrillation and anticoagulation in a community hospital. AB - BACKGROUND: Clinical trials have established the safety and efficacy of warfarin anticoagulation for stroke prevention in patients with atrial fibrillation. Other studies have documented patterns of underutilization and suboptimal warfarin therapy; physician underuse of warfarin may reflect the demands associated with monitoring the drug's effects. BASELINE STUDY: At Carney Hospital, a 230-bed acute care community teaching hospital in Boston, a retrospective chart review indicated that between July 1, 1995, and June 30, 1996, of 465 patients admitted with atrial fibrillation, 209 (45%) patients were discharged with warfarin therapy: 198 were receiving warfarin at admission, and 11 began therapy during hospitalization. Analysis of the admission international normalized ratios (INRs) indicated that a minority of patients on warfarin were safely anticoagulated at the time of admission. DESIGNING THE INTERVENTION: An anticoagulation clinic was established in fall 1997 to increase utilization of warfarin, standardize anticoagulation practices, and minimize physician time and effort needed to ensure safe anticoagulation. In early 1998 monitoring of hospitalized patients with chronic atrial fibrillation began. RESULTS: The proportion of patients receiving warfarin therapy at admission increased from 46% in February-May 1998 to 63% in April-June 1999. Between October 1997 and July 1998, 49.1% of the 2,738 patient visits to the anticoagulation clinic showed an INR in the desired range. For the 2,238 visits during January through August 1999, 53.7% of the INRs were in the desired range. DISCUSSION: Establishment of a clinic to oversee warfarin therapy and dissemination of indications for anticoagulation in patients with atrial fibrillation were followed by increases in the frequency of warfarin use in hospital patients and the incidence of safe therapy in ambulatory patients. PMID- 10677820 TI - Merging clinical pathway programs as part of overall health system mergers: a ten step guide. Spectrum Health. AB - BACKGROUND: Mergers, acquisitions, and reorganizations can be stressful and accompanied by ambivalence, confusion, and uncertainty. Providing clear and simple steps for merging clinical pathways may help organizations move through the transition process more smoothly. The ten steps according to which Spectrum Health merged its pathway program-conduct an inventory of previous efforts, plan for the ideal program, bring staff together early in the merger process, decide on a common format, standardize the development and revision process, standardize a reporting tool, create a clinical pathway manual, implement an educational plan, present the program to key customers, and appoint an advisory group-need not be done sequentially. The ten-step pathway merger program uses pathways as a means to improve the quality of the care provided, with a focus on multidisciplinary clinical pathway teamwork. Before the merger, the two hospital systems' pathway programs used different approaches to operations and pathway format. When the announcement to merge came in September 1997, steps to merge the clinical pathway programs began. DISCUSSION: More than two years into the merger, Spectrum Health continues to struggle with the evolution of the health system. Clinical pathways represent just one of the significant and extensive issues related to organizational mergers; organizational values, finances, vision, mission, customer relations, strategic priorities, and people issues are a few of the others. Focusing on merging programs such as clinical pathway programs can help put one large piece of the merger puzzle in place and reduce some of the ambiguity associated with all mergers. Executive support is critical to the success of the clinical pathway program. PMID- 10677821 TI - Controlled trials of CQI and academic detailing to implement a clinical practice guideline for depression. AB - BACKGROUND: The release of the Agency for Health Care Policy and Research (AHCPR)'s Guideline for the Detection and Treatment of Depression in Primary Care created an opportunity to evaluate under naturalistic conditions the effectiveness of two clinical practice guideline implementation methods: continuous quality improvement (CQI) and academic detailing. A study conducted in 1993-1994 at Kaiser Permanente Northwest Division, a large, not-for-profit prepaid group practice (group-model) HMO, tested the hypotheses that each method would increase the number of members receiving depression treatment and would relieve depressive symptoms. METHODS: Two trials were conducted simultaneously among adult primary care physicians, physician assistants, and nurse practitioners, using the same guideline document, measurement methods, and one year follow-up period. The academic detailing trial was randomized at the clinician level. CQI was assigned to one of the setting's two geographic areas. To account for intraclinician correlation, both trials were evaluated using generalized equations analysis. RESULTS: Most of the CQI team's recommendations were not implemented. Academic detailing increased treatment rates, but--in a cohort of patients with probable chronic depressive disorder--it failed to improve symptoms and reduced measures of overall functional status. CONCLUSIONS: New organizational structures may be necessary before CQI teams and academic detailing can substantially change complex processes such as the primary care of depression. New research and treatment guidelines are needed to improve the management of persons with chronic or recurring major depressive disorder. PMID- 10677822 TI - Water-macromolecule interactions by NMR: a quadrature-free constant-time approach and its application to CI2. AB - A pulse sequence is proposed to select water magnetization with enhanced specificity through a synergetic combination of several filtering principles. This approach relies on a constant-time evolution period implemented without quadrature detection, which results in a square root 2 increase in signal-to noise ratios as compared to traditional non-selective methods for water filtration. In addition, the quadrature-free constant-time block facilitates the implementation of the water flip-back strategy, which leads to further gains in sensitivity. The proposed experiment was applied to unlabeled HEW lysozyme and to 15N-labeled chymotrypsin inhibitor 2 which was partially or non 13C-enriched. Water molecules belonging to a spine of hydration between two pseudo beta-sheet strands were identified, solving previously reported discrepancies between the X ray and refined NMR structure of CI2. The proposed experiment in particularly suitable for hydration studies of mixtures of labeled and unlabeled components, such as ligand-macromolecule complexes. PMID- 10677823 TI - Random-coil chemical shifts of phosphorylated amino acids. AB - The 1H, 13C, 15N and 31P random-coil chemical shifts and phosphate pKa values of phosphorylated amino acids pSer, pThr and pTyr in the protected peptide Ac-Gly Gly-X-Gly-Gly-NH2 have been obtained in water at 25 degrees C over the pH range 2 to 9. Analysis of ROESY spectra indicates that the peptides are unstructured. Phosphorylation induces changes in random-coil chemical shifts, some of which are comparable to those caused by secondary structure formation, and are therefore significant in structural analyses based on the chemical shift. PMID- 10677824 TI - Observation of internucleotide NH...N hydrogen bonds in the absence of directly detectable protons. AB - Several structural motifs found in nucleic acids involve N-H...N hydrogen bonds in which the donor hydrogens are broadened to extinction due to chemical or conformational exchange. In such situations, it is impossible to use the well established HNN-COSY or soft HNN-COSY experiments, which report the presence of the hydrogen bond directly on the donor proton(s). We present a pulse sequence, H(CN)N(H), for alleviating this problem in hydrogen bonds of the type NdH...Na CH, in which the donor Nd nitrogen is correlated with the corresponding non exchangeable C-H proton associated with the acceptor Na nitrogen. In this way, missing NdH...Na correlations in an HNN-COSY spectrum may be recovered from CH-Nd correlations in the H(CN)N(H) spectrum. By correlating a different set of nuclei relative to the HNN-COSY class of experiments, the H(CN)N(H) experiment also serves to remove ambiguities associated with degeneracies in HNN-COSY spectra. The technique is demonstrated on d(GGAGGAG)4,a quadruplex containing a novel A. (G.G.G.G). A hexad and on d(GGGCAGGT)4, containing a G.G.G.C tetrad, in which missing NH2...N7 correlations are retrieved via H8-(N2,N6) correlations in the H(CN)N(H) spectrum. PMID- 10677825 TI - Solvation study of the non-specific lipid transfer protein from wheat by intermolecular NOEs with water and small organic molecules. AB - Intermolecular nuclear Overhauser effects (NOEs) were measured between the protons of various small solvent or gas molecules and the non-specific lipid transfer protein (ns-LTP) from wheat. Intermolecular NOEs were observed with the hydrophobic pocket in the interior of wheat ns-LTP, which grew in intensity in the order cyclopropane (saturated solution) < methane (140 bar) < ethane (40 bar) < acetonitrile (5% in water) < cyclohexane (saturated solution) < benzene (saturated solution). No intermolecular, NOEs were observed with dioxane (5% in water). The intermolecular NOEs were negative for all of the organic molecules tested. Intermolecular NOEs between wheat ns-LTP and water were weak or could not be distinguished from exchange-relayed NOEs. As illustrated by the NOEs with cyclohexane versus dioxane, the hydrophobic pocket in wheat ns-LTP preferably binds non-polar molecules. Yet, polar molecules like acetonitrile can also be accommodated. The pressure dependence of the NOEs between methane and wheat ns LTP indicated incomplete occupancy, even at 190 bar methane pressure. In general, NOE intensities increased with the size of the ligand molecule and its vapor pressure. NMR of the vapor phase showed excellent resolution between the signals from the gas phase and those from the liquid phase. The vapor concentration of cyclohexane was fivefold higher than that of the dioxane solution, supporting the binding of cyclohexane versus uptake of dioxane. PMID- 10677826 TI - Efficient side-chain and backbone assignment in large proteins: application to tGCN5. AB - In determining the structure of large proteins by NMR, it would be desirable to obtain complete backbone, side-chain, and NOE assignments efficiently, with a minimum number of experiments and samples. Although new strategies have made backbone assignment highly efficient, side-chain assignment has remained more difficult. Faced with the task of assigning side-chains in a protein with poor relaxation properties, the Tetrahymena histone acetyltransferase tGCN5, we have developed an assignment strategy that would provide complete side-chain assignments in cases where fast 13C transverse relaxation causes HCCH-TOCSY experiments to fail. Using the strategy presented here, the majority of aliphatic side-chain proton and carbon resonances can be efficiently obtained using optimized H(CC-CO)NH-TOCSY and (H)C(C-CO)NH-TOCSY experiments on a partially deuterated protein sample. Assignments can be completed readily using additional information from a 13C-dispersed NOESY-HSQC spectrum. Combination of these experiments with H(CC)NH-TOCSY and (H)C(C)NH-TOSCY may provide complete backbone and side-chain assignments for large proteins using only one or two samples. PMID- 10677827 TI - Determination of sugar conformation in large RNA oligonucleotides from analysis of dipole-dipole cross correlated relaxation by solution NMR spectroscopy. AB - A new experiment, the forward directed quantitative gamma-HCCH-TOCSY for the measurement of the conformation of the five-membered ribosyl unit in RNA oligonucleotides, is presented. The experiment relies on quantification of cross peak intensities caused by evolution of CH, CH-dipole-dipole cross correlated relaxation in non-evolution periods and the resolution enhancement obtainable in forward directed HCC-TOCSY transfer. Cross correlated relaxation rates are interpreted to reveal the sugar conformation of 22 out of 25 nucleotides in an isotopically labelled 25-mer RNA. The results obtained with this new method are in agreement with the conformational analysis derived from 3J(H,H) coupling constants. PMID- 10677829 TI - 1H and 15N sequential assignment and solution secondary structure of 15N labelled human pancreatic ribonuclease. PMID- 10677828 TI - HYPER: a hierarchical algorithm for automatic determination of protein dihedral angle constraints and stereospecific C beta H2 resonance assignments from NMR data. AB - A new computer program, HYPER, has been developed for automated analysis of protein dihedral angle values and C beta H2 stereospecific assignments from NMR data. HYPER uses a hierarchical grid-search algorithm to determine allowed values of phi, psi, and chi 1 dihedral angles and C beta H2 stereospecific assignments based on a set of NMR-derived distance and/or scalar-coupling constraints. Dihedral-angle constraints are valuable for restricting conformational space and improving convergence in three-dimensional structure calculations. HYPER computes the set of phi, psi, and chi 1 dihedral angles and C beta H2 stereospecific assignments that are consistent with up to nine intraresidue and sequential distance bounds, two pairs of relative distance bounds, thirteen homo- and heteronuclear scalar coupling bounds, and two pairs of relative scalar coupling constant bounds. The program is designed to be very flexible, and provides for simple user modification of Karplus equations and standard polypeptide geometries, allowing it to accommodate recent and future improved calibrations of Karplus curves. The C code has been optimized to execute rapidly (0.3-1.5 CPU-sec residue-1 using a 5 degrees grid) on Silicon Graphics R8000, R10000 and Intel Pentium CPUs, making it useful for interactive evaluation of inconsistent experimental constraints. The HYPER program has been tested for internal consistency and reliability using both simulated and real protein NMR data sets. PMID- 10677830 TI - 1H, 13C and 15N chemical shift assignments of the capsid protein from Rous sarcoma virus. PMID- 10677831 TI - 1H, 15N, and 13C resonance assignment of the PH domain from C. elegans UNC-89. PMID- 10677832 TI - The use of solid-phase microextraction in conjunction with a benchtop quadrupole mass spectrometer for the analysis of volatile organic compounds in human blood at the low parts-per-trillion level. AB - The analysis of volatile organic compounds (VOCs) in whole human blood at the low parts-per-trillion level has until recently required the use of a high-resolution mass spectrometer to obtain the specificity and detection limits required for epidemiological studies of VOC exposure in the general public. Because of the expense and expertise required to operate and maintain a high-resolution instrument, the applicability of this method has been limited. These limitations are overcome in a new method using automated headspace solid-phase microextraction (SPME) in conjunction with a gas chromatograph and a benchtop quadrupole mass spectrometer. A combination of SPME and multiple single-ion monitoring minimizes the interferences and chemical noise associated with whole blood samples. This method permits the analysis of 10 VOCs in human blood while simplifying the sample preparation and reducing the possible exposure of the analyst to blood aerosols. Twelve samples can be run successively in a fully automated mode, thus eliminating the need for operator attention. Detection limits are below 50 ppt (pg/mL) for a majority of the VOCs tested with a 5-mL sample. PMID- 10677833 TI - Determination of polycyclic aromatic hydrocarbons in sediment using solid-phase microextraction with gas chromatography-mass spectrometry. AB - Manual solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) is applied for the determination of polycyclic aromatic hydrocarbons (PAHs) from natural matrix through a distilled water medium. Seven of the 16 PAH standards (naphthalene, acenaphthene, fluorene, anthracene, fluoranthene, pyrene, benzo[a]anthracene) are spiked on a marine muddy sediment. The samples, containing PAHs in the range of 10-20 ppm, are then aged at room temperature more than 10 days before analysis. The influence of the matrix, SPME adsorption time, pH, salt content, and SPME adsorption temperature are investigated. The reproducibility of the technique is less than 13% (RDS) for the first 6 considered PAHs and 28% (RDS) for benzo(a)anthracene with a fiber containing a 100-micron poly dimethylsiloxane coating. Linearity extended in the range of 5-50 picograms for PAHs direct injection, 5-70 picograms for PAHs in water, and 1-170 picograms for PAHs in sediment. The detection limit is estimated less than 1 microgram/kg of dry sample for the first 6 considered PAHs in sediment and 1.5 micrograms/kg of dry sample for benzo(a)anthracene using the selected ion monitoring mode in GC-MS. The recoveries of the considered PAHs are evaluated. PMID- 10677834 TI - Complexation ion-exchange chromatography of some metal ions on papers impregnated with Ti(IV)-based inorganic ion exchangers. AB - The chromatographic behavior of 40 metal ions is studied on titanium (IV) arsenate, titanium (IV) phosphate-, titanium (IV) molybdate-, titanium(IV) tungstate-, and titanium(IV) selenite-impregnated papers in 0.1M oxalic, citric, and tartaric acid as mobile phases. Similar studies are carried out on Whatman No. 1 papers for comparison. The ion-exchange capacity of these papers is determined, and their selectivity for different cations is discussed. The mechanism of migration is explained in terms of ion-exchange, precipitation, and adsorption. The prediction of elution sequence from RF values is also checked. The average Ri is found to be almost linearly dependent on the charge of the metal ions. The effect of the pKa of complexing acids on average RF values of 3d series metal ions is explained. A number of binary and ternary separations are achieved. PMID- 10677835 TI - A convenient method for routine estimation of dead time in gas chromatography. AB - A set of retention times (tR) of n-alkanes at different temperatures and the primary dead times (tM) are used to determine the 4 numerical constants (a, b, c, and d) of an equation. This equation is rearranged into a second equation, used in turn for routine calculation of the secondary dead time (tMS) of each chromatogram from any member of the n-alkane series. Equation 2 can be used to calculate the tMS of both packed and capillary columns. The calculated tMS values are in good agreement with those of the tM values. The greatest difference is 3.38%, but it is speculated that the percent difference would be lower when more tR data are collected for the determination of the 4 numerical constants of Equation 1. Error in measurement of retention time undoubtedly would affect the accuracy of the estimated dead time, but it is attenuated by a factor of 1 + e(a + bN + c/T + dN/T). PMID- 10677836 TI - Liquid chromatographic determination of rifampin in human plasma, bronchoalveolar lavage fluid, and alveolar cells. AB - A technique is presented for the measurement of rifampin in humans by reversed phase column chromatography and postcolumn photo irradiation. The assay employs diazepam as an internal standard and provides specific, rapid, and reliable determinations for drug concentration in plasma, bronchoalveolar lavage (BAL), and alveolar cells (AC). The preparation of plasma and AC samples utilizes a simple deproteinization step, whereas BAL supernatants require a solid-phase extraction. The assay produces sharp peaks with retention times of 6.2 and 15 min for rifampin and diazepam, respectively. The detection limits for rifampin were 0.5 microgram/mL for plasma, 0.015 microgram/mL for BAL supernatants, and 0.03 microgram/mL for AC suspensions. The assay has excellent performance characteristics, making it suitable for pharmacological studies in humans, and it is being used to support a study of the intrapulmonary pharmacokinetics of rifampin. PMID- 10677838 TI - Gas chromatography problem solving and troubleshooting. PMID- 10677837 TI - Analyte loss due to membrane filter adsorption as determined by high-performance liquid chromatography. AB - The phenomenon of membrane filter adsorption in high-performance liquid chromatography (HPLC) is investigated utilizing 16 brands of filters representing 3 polymeric materials: cellulose acetate (CA), nylon, and polyvinylidene difluoride in a variety of diameters (3, 4, 7, 13, and 25 mm). Sixteen compounds commonly encountered in drug preparations are selected as sample analytes and classified as acidic, basic, and neutral in chemical behavior. Six mobile phase/sample solvent mixtures are included: 3 with methanol-water and 3 with acetonitrile-water as major constituents. When using methanol as the mobile phase organic component, CA, nylon, and polyvinylidene difluoride (PVDF) filters exhibit negligible to moderate adsorption levels with regard to the neutral and basic drug compounds. The acidic drug test compounds are adsorbed by 50% of all 3 filter materials tested in methanol-water. In acetonitrile, neutral compounds are affected by 31.4%, basic compounds are affected by 47.0%, and acidic compounds are affected by 53.6% of the nylon and PVDF filters. CA is incompatible with acetonitrile and is excluded from the study with this solvent. PMID- 10677839 TI - Disinfectant effects of hot water, ultraviolet light, silver ions and chlorine on strains of Legionella and nontuberculous mycobacteria. AB - The disinfectant effects on Legionella and nontuberculous mycobacteria of hot water, ultraviolet light, silver ions and chlorine, were evaluated. The bacterial strains Legionella pneumophila ATCC33152 and Mycobacterium avium ATCC25291 and strains of L. pneumophila and M. avium which had been isolated from a 24 h bath, were examined for their resistance to treatments. All strains were killed within 3 min on exposure to hot water at 70 degrees C and exposure to ultraviolet light at 90 mW.s/cm2. The strains of L. pneumophila tested were killed within 6 h on exposure to a solution of silver ions at 50 micrograms/l. The number of viable cells of strains of M. avium fell from 10(5) CFU/ml to 10(3) CFU/ml after exposure to an aqueous solution of silver ions at 100 micrograms/l for 24 h. Chlorine effectively killed strains of Legionella which were exposed to an aqueous solution of chlorine at 2 mg/l within 3 min, but strains of Mycobacterium survived exposure to chlorine at 4 mg/l for more than 60 min. PMID- 10677840 TI - Enhancement of growth and toxin production of Clostridium argentinense by co culture with Pseudomonas mendocina. AB - The growth and toxin production of Clostridium argentinense in co-culture with Pseudomonas mendocina was examined in a micro-fermenter without aeration characterizing the association in terms of several growth parameters. The biomass obtained in co-cultures was 4.4 times higher than that in C. argentinense monocultures with total consumption of the carbon source. The pH and Eh attained in co-cultures at later stages of cultivation were suitable for toxin production by C. argentinense. In comparison with C. argentinense monocultures the production of toxin was 17.5 times higher with a specific toxicity of 0.56 LD50 per g of co-culture biomass. PMID- 10677841 TI - Flow cytometric analysis of a marine LAS-degrading consortia. AB - The specific nucleic acid fluorochrome SYTO-13 was used in flow cytometric analysis to assess changes in the density and heterogeneity of marine bacterial populations which biodegrade linear alkylbenzene sulphonate (LAS). Seawater samples with LAS and incubated in the laboratory (20 degrees C, 100 rpm, 30 days) were used to monitor LAS-degrading consortia. Flow cytometric studies and culture methods were used to characterize the LAS degrading bacterioplankton consortia. Fluorescence and scatter signals enabled us to define three regions (R1, R2 and R3) in the dual parameter cytograms. The distribution of the bacterial counts in these regions allowed us to monitor the formation and evolution of the consortia. PMID- 10677842 TI - Effects of caffeic acid, chlorogenic acid and ferulic acid on growth and arylamine N-acetyltransferase activity in Shigella sonnei (group D). AB - Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene (2-AF) as substrates were determined in Shigella sonnei (group D) collected from patients with diarrhoeal disease. The NAT activity was determined using an acetyl CoA recycling assay and high pressure liquid chromatography. Inhibition of growth studies from S. sonnei (group D) demonstrated that caffeic acid (CA), chlorogenic acid (CGA) and ferulic acid (FA) elicited a dose-dependent bactericidal effect in S. sonnei (group D) cultures, i.e. the greater the concentration of CA, CGA and FA, the greater the inhibition of growth of S. sonnei (group D). Cytosols or suspensions of S. sonnei (group D) with and without selected concentrations of CA, CGA and FA co-treatment showed different percentages of 2-AF acetylation. The data indicated that there was reduced NAT activity associated with increased CA, CGA and FA in Shigella dysenteriae (group D) cytosols and intact cells. For the cytosol and intact bacteria examinations, the apparent values of K(m) and Vmax decreased after being co-treated with 400 microM CA, CGA and FA. This report is the first demonstration of plant phenolic inhibition (CA, CGA and FA) of arylamine NAT activity and growth in the bacterium S. sonnei (group D). PMID- 10677843 TI - Effects of media composition of delta-endotoxin production and morphology of Bacillus thuringiensis in wild types and spontaneously mutated strains. AB - Bacillus thuringiensis strains HD-73 and 4412, and two spontaneous mutants termed 4412aa-ind and 4412sph-cry were studied for the ability to produce crystals of different size and shape when grown in a rich medium and in an appropriate minimal medium defined during this study. Strain 4412aa-ind showed medium dependent variation in the crystal phenotype. Scanning electron microscopy was utilized in order to show crystal variations in size and shape. B. thuringiensis strains 4412aa-ind and 4412sph-cry grown in rich and in minimal media produced differences in crystal morphology, and SDS-PAGE gel indicated that crystal variation was only at the morphological level and not in composition of the amino acids. A further nineteen B. thuringiensis strains were tested for the ability to sporulate in two simple defined media. Of these strains thirteen were able to complete sporulation with crystal production in one or both media. All strains grew and sporulated in a medium containing the usual twenty amino acids, and no vitamins or other growth factors were required. PMID- 10677844 TI - Compartment-specific isoforms of TPI and GAPDH are imported into diatom mitochondria as a fusion protein: evidence in favor of a mitochondrial origin of the eukaryotic glycolytic pathway. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and triosephosphate isomerase (TPI) are essential to glycolysis, the major route of carbohydrate breakdown in eukaryotes. In animals and other heterotrophic eukaryotes, both enzymes are localized in the cytosol; in photosynthetic eukaryotes, GAPDH and TPI exist as isoenzymes that function in the glycolytic pathway of the cytosol and in the Calvin cycle of chloroplasts. Here, we show that diatoms--photosynthetic protists that acquired their plastids through secondary symbiotic engulfment of a eukaryotic rhodophyte--possess an additional isoenzyme each of both GAPDH and TPI. Surprisingly, these new forms are expressed as an TPI-GAPDH fusion protein which is imported into mitochondria prior to its assembly into a tetrameric bifunctional enzyme complex. Homologs of this translational fusion are shown to be conserved and expressed also in nonphotosynthetic, heterokont-flagellated oomycetes. Phylogenetic analyses show that mitochondrial GAPDH and its N-terminal TPI fusion branch deeply within their respective eukaryotic protein phylogenies, suggesting that diatom mitochondria may have retained an ancestral state of glycolytic compartmentation that existed at the onset of mitochondrial symbiosis. These findings strongly support the view that nuclear genes for enzymes of glycolysis in eukaryotes were acquired from mitochondrial genomes and provide new insights into the evolutionary history (host-symbiont relationships) of diatoms and other heterokont-flagellated protists. PMID- 10677845 TI - Mitochondrial genes and mammalian phylogenies: increasing the reliability of branch length estimation. AB - Since branch lengths provide important information about the timing and the extent of evolutionary divergence among taxa, accurate resolution of evolutionary history depends as much on branch length estimates as on recovery of the correct topology. However, the empirical relationship between the choice of genes to sequence and the quality of branch length estimation remains ill defined. To address this issue, we evaluated the accuracy of branch lengths estimated from subsets of the mitochondrial genome for a mammalian phylogeny with known subordinal relationships. Using maximum-likelihood methods, we estimated branch lengths from an 11-kb sequence of all 13 protein-coding genes and compared them with estimates from single genes (0.2-1.8 kb) and from 7 different combinations of genes (2-3.5 kb). For each sequence, we separated the component of the log likelihood deviation due to branch length differences associated with alternative topologies from that due to those that are independent of the topology. Even among the sequences that recovered the same tree topology, some produced significantly better branch length estimates than others did. The combination of correct topology and significantly better branch length estimation suggests that these gene combinations may prove useful in estimating phylogenetic relationships for mammalian divergences below the ordinal level. Thus, the proper choice of genes to sequence is a critical factor for reliable estimation of evolutionary history from molecular data. PMID- 10677846 TI - High DNA sequence variability at the alpha 1 Na/K-ATPase locus of Artemia franciscana (brine shrimp): polymorphism in a gene for salt-resistance in a salt resistant organism. AB - We previously reported that the Na/K-ATPase alpha 1 subunit coding gene showed signs of being a very polymorphic locus in Artemia franciscana. This species is adapted to highly saline waters, and the Na/K-ATPase alpha 1 isoform presumably plays a key role in this adaptation. Therefore, we were interested in further study of the alpha 1 Na/K-ATPase polymorphisms to examine whether they might be due to an adaptation to salt resistance driven by natural selection. Using coding sequences from 10 genomic clones and 3 cDNAs, we observed that most substitutions are in synonymous positions (88.8%). The 12 nonsynonymous substitutions code for conservative amino acid replacements with an apparent scattered distribution across functional domains of the protein. Interspecific comparison between these sequences and two genomic clones from Artemia parthenogenetica containing 1,122 bp of the alpha 1 Na/K-ATPase locus coding sequence showed independence of the synonymous/nonsynonymous ratio in the comparison within A. franciscana and between A. franciscana and A. parthenogenetica, which fits the neutral model of evolution. Since there were no previous studies on DNA polymorphism for other A. franciscana genes, we also studied variability at the Actin 302 locus for comparison. Both loci were amplified by reverse transcription-polymerase chain reaction, and 20 sequences were obtained for each. This study shows that the amplified region of the alpha 1 Na/K-ATPase gene is 3.5 times as polymorphic as the Actin 302 gene and 2.9 times as heterozygotic. Interestingly, under a model of neutral evolution, the data observed would be expected with a probability of approximately 0.05, suggesting an excess of intraspecific variation of alpha 1 Na/K-ATPase with respect to Actin 302. Restriction fragment length polymorphism studies show similar patterns of polymorphism along the approximately 41-kb span of the alpha 1 Na/K-ATPase locus. Most of the nucleotide differences are linked in a few haplotypes, although recombination events are also inferred from the data. We propose a possible explanation for the high polymorphic levels at the alpha 1 Na/K-ATPase locus which invokes positive selection acting tightly to the locus in transiently isolated or semi-isolated subpopulations. PMID- 10677847 TI - Multiple acquisition of methanogenic archaeal symbionts by anaerobic ciliates. AB - Anaerobic heterotrichous ciliates (Armophoridae and Clevelandellidae) possess hydrogenosomes that generate molecular hydrogen and ATP. This intracellular source of hydrogen provides the basis for a stable endosymbiotic association with methanogenic archaea. We analyzed the SSU rRNA genes of 18 heterotrichous anaerobic ciliates and their methanogenic endosymbionts in order to unravel the evolution of this mutualistic association. Here, we show that the anaerobic heterotrichous ciliates constitute at least three evolutionary lines. One group consists predominantly of gut-dwelling ciliates, and two to three, potentially four, additional clades comprise ciliates that thrive in freshwater sediments. Their methanogenic endosymbionts belong to only two different taxa that are closely related to free-living methanogenic archaea from the particular ecological niches. The close phylogenetic relationships between the endosymbionts and free-living methanogenic archaea argue for multiple acquisitions from environmental sources, notwithstanding the strictly vertical transmission of the endosymbionts. Since phylogenetic analysis of the small-subunit (SSU) rRNA genes of the hydrogenosomes of these ciliates indicates a descent from the mitochondria of aerobic ciliates, it is likely that anaerobic heterotrichous ciliates hosted endosymbiotic methanogens prior to their radiation. Therefore, our data strongly suggest multiple acquisitions and replacements of endosymbiotic methanogenic archaea during their host's adaptation to the various ecological niches. PMID- 10677848 TI - Rapid expansion of microsatellite sequences in pines. AB - Microsatellite persistence time and evolutionary change was studied among five species of pines, which included a pair of closely related species (Pinus sylvestris and Pinus resinosa) in the subgenus Pinus, their relative Pinus radiata, and another closely related species pair (Pinus strobus and Pinus lambertiana) in the subgenus Strobus. The effective population sizes of these species are known to have ranged from the very small bottlenecks of P. resinosa to vast populations of P. sylvestris. This background allowed us to place the microsatellite evolution in a well-defined phylogenetic setting. Of 30 loci originating from P. strobus and P. radiata, we were able to consistently amplify 4 in most of the these pine species. These priming sites had been conserved for over 100 Myr. The four microsatellites were sequenced in the five species. Flanking sequences were compared to establish that the loci were orthologous. All microsatellites had persisted in these species, despite very different population sizes. We found a recent microsatellite duplication: a closely related pair of loci in P. strobus, where the other four species had just one locus. On two independent occasions, the repeat area of this same microsatellite (locus RPS 105a/b) had grown from a very low repeat number to 15 or 17 in the last 10-25 Myr. Other parts of the same compound microsatellite had remained virtually unchanged. Locus PR 4.6 is known to be polymorphic in both P. radiata and P. sylvestris, but the polymorphism in the two species is due to different motifs. The very large pine genomes are highly repetitive, and microsatellite loci also occur as gene families. PMID- 10677849 TI - Complete sequence of the mitochondrial DNA of the primitive opisthobranch gastropod Pupa strigosa: systematic implication of the genome organization. AB - The complete sequence (14,189 bp) of the mitochondrial DNA of the opisthobranch gastropod Pupa strigosa was determined. The genome contains 13 protein, 2 rRNA, and 22 tRNA genes typical of metazoan mtDNA. The Pupa mitochondrial genome is highly compact and shows the following unusual features, like pulmonate land snails: (1) extremely small genome size, (2) absence of lengthy noncoding regions (with the largest intergenic spacer being only 46 nt), (3) size reduction of encoded genes, and (4) many overlapping genes. Several tRNA genes exhibit bizarre secondary structures with reduced T or D stems, and many tRNA genes have unstable acceptor stems that might be corrected by posttranscriptional RNA editing. The Pupa mitochondrial gene arrangement is almost identical to those of pulmonate land snails but is radically divergent from those of the prosobranch gastropod Littorina saxatilis and other molluscs. Our finding that the unique gene arrangement and highly compact genome organization are shared between opisthobranch and pulmonate gastropods strongly suggests their close phylogenetic affinity. PMID- 10677850 TI - An unusual form of purifying selection in a sperm protein. AB - Protamines are small, highly basic DNA-binding proteins found in the sperm of animals. Interestingly, the proportion of arginine residues in one type of protamine, protamine P1, is about 50% in mammals. Upon closer examination, it was found that both the total number of amino acids and the positions of arginine residues have changed considerably during the course of mammalian evolution. This evolutionary pattern suggests that protamine P1 is under an unusual form of purifying selection, in which the high proportion of arginine residues is maintained but the positions may vary. In this case, we would expect that the rate of nonsynonymous substitution is not particularly low compared with that of synonymous substitution, despite purifying selection. We would also expect that the selection for a high arginine content results in a high frequency of the nucleotide G in the coding region of this gene, because all six arginine codons contain at least one G. These expectations were confirmed in our study of mammalian protamine genes. Analysis of nonmammalian vertebrate genes also showed essentially the same patterns of evolutionary changes, suggesting that this unusual form of purifying selection has been active since the origin of bony vertebrates. The protamine gene of an insect species shows similar patterns, although its purifying selection is less intense. These observations suggest that arginine-rich selection is a general feature of protamine evolution. The driving force for arginine-rich selection appears to be the DNA-binding function of protamine P1 and an interaction with a protein kinase in the fertilized egg. PMID- 10677851 TI - Intragenomic variation within ITS1 and ITS2 of freshwater crayfishes (Decapoda: Cambaridae): implications for phylogenetic and microsatellite studies. AB - Intragenomic variation in ITS1 and ITS2 is known to exit but is widely ignored in phylogenetic studies using these gene regions. The amount of variation in seven crayfish species, including three populations of Orconectes luteus and two of Procambarus clarkii, was assessed by sequencing 3, 5, or 10 clones from the same individuals, for a total of 77 sequences. The ITS1 and ITS2 sequences reported here are some of the longest known, with aligned lengths of 760 and 1,300 bp, respectively. They contain multiple microsatellite insertions, all of which show considerable intragenomic variation in the number of repeat elements. This variation is enough to obscure phylogenetic relationships at the population level, although relationships between species can be estimated. Given the hybridization techniques used to locate microsatellites, multiple-copy regions like ITS1 and ITS2 will be preferentially found if they contain microsatellites, and in these cases the microsatellites will not behave as typical Mendelian markers and could give spurious results. PMID- 10677852 TI - The alpha-mannosidases: phylogeny and adaptive diversification. AB - alpha-Mannosidase enzymes comprise a class of gylcoside hydrolases involved in the maturation and degradaton of glycoprotein-linked oligosaccharides. Various alpha-mannosidase enzymatic activities are encoded by an ancient and ubiquitous gene superfamily. A comparative sequence analysis was employed here to characterize the evolutionary relationships and dynamics of the alpha-mannosidase superfamily. A series of lineage-specific BLAST searches recovered the first ever recognized archaean and eubacterial alpha-mannosidase sequences, in addition to numerous eukaryotic sequences. Motif-based alignment and subsequent phylogenetic analysis of the entire superfamily revealed the presence of three well-supported monophyletic clades that represent discrete alpha-mannosidase families. The comparative method was used to evaluate the phylogenetic distribution of alpha mannosidase functional variants within families. Results of this analysis demonstrate a pattern of functional diversification of alpha-mannosidase paralogs followed by conservation of function among orthologs. Nucleotide polymorphism among the most closely related pair of duplicated genes was analyzed to evaluate the role of natural selection in the functional diversification of alpha mannosidase paralogs. Ratios of nonsynonymous and synonymous variation show an increase in the rate of nonsynonymous change after duplication and a relative excess of fixed nonsynonymous changes between the two groups of paralogs. These data point to a possible role for positive Darwinian selection in the evolution of alpha-mannosidase functional diversification following gene duplication. PMID- 10677853 TI - Solvent accessibility and purifying selection within proteins of Escherichia coli and Salmonella enterica. AB - The neutral theory of molecular evolution predicts that variation within species is inversely related to the strength of purifying selection, but the strength of purifying selection itself must be related to physical constraints imposed by protein folding and function. In this paper, we analyzed five enzymes for which polymorphic sequence variation within Escherichia coli and/or Salmonella enterica was available, along with a protein structure. Single and multivariate logistic regression models are presented that evaluate amino acid size, physicochemical properties, solvent accessibility, and secondary structure as predictors of polymorphism. A model that contains a positive coefficient of association between polymorphism and solvent accessibility and separate intercepts for each secondary structure element is sufficient to explain the observed variation in polymorphism between sites. The model predicts an increase in the probability of amino acid polymorphism with increasing solvent accessibility for each protein regardless of physicochemical properties, secondary-structure element, or size of the amino acid. This result, when compared with the distribution of synonymous polymorphism, which shows no association with solvent accessibility, suggests a strong decrease in purifying selection with increasing solvent accessibility. PMID- 10677854 TI - The glutamine synthetases of rhizobia: phylogenetics and evolutionary implications. AB - Glutamine synthetase exists in at least two related forms, GSI and GSII, the sequences of which have been used in evolutionary molecular clock studies. GSI has so far been found exclusively in bacteria, and GSII has been found predominantly in eukaryotes. To date, only a minority of bacteria, including rhizobia, have been shown to express both forms of GS. The sequences of equivalent internal fragments of the GSI and GSII genes for the type strains of 16 species of rhizobia have been determined and analyzed. The GSI and GSII data sets do not produce congruent phylogenies with either neighbor-joining or maximum likelihood analyses. The GSI phylogeny is broadly congruent with the 16S rDNA phylogeny for the same bacteria; the GSII phylogeny is not. There are three striking rearrangements in the GSII phylograms, all of which might be explained by horizontal gene transfer to Bradyrhizobium (probably from Mesorhizobium), to Rhizobium galegae (from Rhizobium), and to Mesorhizobium huakuii (perhaps from Rhizobium). There is also evidence suggesting intrageneric DNA transfer within Mesorhizobium. Meta-analysis of both GS genes from the different genera of rhizobia and other reference organisms suggests that the divergence times of the different rhizobium genera predate the existence of legumes, their host plants. PMID- 10677855 TI - Evolutionary history of the human endogenous retrovirus family ERV9. AB - Several distinct families of endogenous retrovirus-like elements (ERVs) exist in the genomes of primates. Despite the important evolutionary consequences that carrying these intragenomic parasites may have for their hosts, our knowledge about their evolution is still scarce. A matter of particular interest is whether evolution of ERVs occurs via a master lineage or through several lineages coexisting over long periods of time. In this work, the paleogenomic approach has been applied to the study of the evolution of ERV9, one of the human endogenous retrovirus families mobilized during primate evolution. By searching the GenBank database with the first 676 bp of the ERV9 long terminal repeat, we identified 156 different element insertions into the human genome. These elements were grouped into 14 subfamilies based on several characteristic nucleotide differences. The age of each subfamily was roughly estimated based on the average sequence divergence of its members from the subfamily consensus sequence. Determination of the sequential order of diagnostic substitutions led to the identification of four distinct lineages, which retained their capacity of transposition over extended periods of evolution. Strong evidence for mosaic evolution of some of these lineages is presented. Taken altogether, the available data indicate that the possibility of ERV9 still being active in the human lineage can not be discarded. PMID- 10677856 TI - Evidence for genomic duplication of the glutathione transferase A3 gene in genus Rattus. PMID- 10677857 TI - Neuromuscular disorders in childhood: a descriptive epidemiological study from western Sweden. AB - A retrospective epidemiological study of neuromuscular disorders was carried out in children born between 1979 and 1994 in western Sweden. The purpose was to determine overall and specific prevalences, overall cumulative incidence and birth incidences of selected disorders. Cases were ascertained from 12 different sources and medical records, investigations and diagnosis were reviewed. We found a point prevalence in the population < 16 years of age of 63.1 x 10(-5) for all neuromuscular disorders and 53.1 x 10(-5) for inherited neuromuscular disorders. The point prevalence in children of school age was even higher. We found a higher occurrence of hereditary motor and sensory neuropathy, congenital myopathies and mitochondrial encephalo-myopathy, a slightly lower occurrence of Duchenne muscular dystrophy and spinal muscular atrophy and equal occurrence of myotonic dystrophy compared to previous studies in other countries. We conclude that neuromuscular disorders are more common in childhood than has previously been reported. PMID- 10677858 TI - Autosomal recessive hereditary neuropathy with focally folded myelin sheaths and linked to chromosome 11q23: a distinct and homogeneous entity. AB - We describe a six generation Saudi kindred, with a recessive hereditary motor and sensory neuropathy (HMSN). Four individuals were affected including two children (a boy and a girl) and a 23-year-old man. The fourth (a female) died at the age of 14 years. Onset of the disease was early (< 2 years) and the clinical and neurophysiological features were, generally, quite similar to those of an Italian family linked to chromosome 11q23. The peculiar pathologic pattern was irregular and redundant loops associated with folding of the myelin sheaths. The genetic study confirmed linkage to chromosome 11q23 and refined the location of the gene between D11S1311 and D11S917, a 3.3 cM region. These findings support the existence of a homogeneous and distinct entity within the form of HMSN associated with focally folded myelin sheaths. PMID- 10677859 TI - Muscle membrane-skeleton protein changes and histopathological characterization of muscle-eye-brain disease. AB - Muscle-eye-brain disease belongs to congenital muscular dystrophies with central nervous system abnormalities. The etiology of MEB is still unknown, but abnormal immunoreactivity for laminin-2 has been reported. To evaluate disease progression in muscle tissue, 32 biopsy specimens from 17 muscle-eye-brain patients were analysed. The samples of four patients were studied by immunohistochemical techniques and by quantitative Western blotting. The samples showed a great variation in the muscle pathology. Regenerative fibers and mild fiber size variation were present in over 60%. At infancy, necrotic and regenerative fibers were common, while fat infiltration was the most prominent finding in the age group over five years. In quantitative studies, the amount of laminin alpha 2 chain was clearly reduced to 10-20% of normal. In contrast, laminin beta 2 chain was overexpressed in the Western blotting studies. These findings may reflect a yet unidentified primary disturbance in the basement membrane composition and function. PMID- 10677860 TI - Two distal mutations in the gene encoding emerin have profoundly different effects on emerin protein expression. AB - Emerin, the product of the gene responsible for X-linked Emery-Dreifuss muscular dystrophy (EDMD), has a ubiquitous tissue distribution and is localised to the nuclear envelope. We present here the relationship between emerin protein expression, nuclear localization and clinical phenotype for two distal mutations identified in unrelated EDMD patients. The first mutation predicts the replacement of the last eight amino acids of emerin with the addition of 101 amino acids, but no emerin expression is detected. The second mutation, 35 bp upstream from the first mutation, deletes six amino acids from the transmembrane region, but in this case emerin expression is seen. Emerin from this second patient is expressed at reduced levels, mistargeted and has altered biochemical properties compared to wild type emerin. In both cases the clinical phenotype was similar to patients with typical null mutations. We discuss these data in comparison with previous reports of other C-terminal mutations in the emerin gene and suggest that the efficiency of emerin's nuclear membrane localization is affected by the hydrophobicity (and possibly length) of its transmembrane region, and a longer C-terminal tail prevents nuclear localization. PMID- 10677861 TI - Different effects of mexiletine on two mutant sodium channels causing paramyotonia congenita and hyperkalemic periodic paralysis. AB - Effects of the antiarrhythmic and antimyotonic drug mexiletine were studied on two sodium channel mutants causing paramyotonia congenita (R1448H) and an overlap paramyotonic and hyperkalemic paralytic syndrome (M1360V). Channels were expressed in human embryonic kidney cells and studied electrophysiologically, using the whole-cell patch-clamp technique. Compared to the wild-type, channel, both mutants showed alterations of inactivation, i.e. slower inactivation, left shift of steady-state inactivation and faster recovery from inactivation. Mexiletine caused a significantly larger use-dependent block of the R1448H mutant when compared to M1360V and wild-type channels. This can be explained by a prolonged recovery from mexiletine block as observed for R1448H channels, since the affinity of mexiletine for the inactivated state was similar for all three clones. The use-dependent block of sodium channels by mexiletine reduces repetitive series of action potentials and therefore improves muscle stiffness in myotonic patients. The enhanced use-dependent block as seen with R1448H may explain the extraordinary therapeutic efficacy of mexiletine in most patients with paramyotonia congenita. PMID- 10677862 TI - Effects of aerobic training on lactate and catecholaminergic exercise responses in mitochondrial myopathies. AB - The aim of this study was to evaluate the effects of an aerobic training program on the metabolic and sympathetic responses to exercise in 12 patients with mitochondrial myopathies. A 10-week course of aerobic training, consisting of supervised exercise every other day on an electrically braked pedal-rate bicycle ergometer was prescribed to each patient and four healthy controls. Venous lactate, epinephrine (EP) and norepinephrine (NEP) levels were assessed at baseline and after the aerobic training by means of constant-workload exercise performed at near lactate threshold (LT). In patients, a decrease in exercise peak values, significant for lactate (-38.6%, P < 0.01) but not for catecholamines (EP: -26.0%, NEP: -22.1%) was observed after training, findings confirmed by the lactate/EP and lactate/NEP area ratios. The results show that lactate accumulation during exercise is decreased after aerobic training in mitochondrial myopathies and that the effect is partially dissociated from the catecholaminergic response. This in turn suggests that the lactate decrease can be explained, at least in part, by the improved muscle oxidative metabolism consequent to the proposed training program. PMID- 10677863 TI - Dermatomyositis and Whipple's disease. AB - A 14-year-old boy presented with a 3-year history of a skin rash typical of juvenile dermatomyositis, and a 2-month history of mild proximal weakness, myalgia, and weight loss. A quadriceps biopsy showed perifascicular fibre atrophy, focal necrosis and regeneration, immunohistochemical labelling for HLA-1 on the surface of the fibres, and focal C5-9 deposition in capillaries. Macrophages with diastase-resistant, PAS-positive cytoplasm were present. Ultrastructural studies showed electron dense and membranous debris. The patient's symptoms responded to intravenous immunoglobulin and oral prednisolone. Four months after discontinuing prednisolone, the patient developed cardiac failure, ventricular tachycardia, and a recurrence of his rash. The 16S ribosomal RNA specific for Tropheryma whippelii was identified by polymerase chain reaction (PCR) analysis in skeletal and cardiac muscle. The myalgia and skin rash responded to prednisolone and oral co-trimoxazole, and the tachycardia is controlled by oral verapamil. This patient appears to have a novel association of juvenile dermatomyositis and Whipple's disease. PMID- 10677864 TI - An Italian family with Ala-47 transthyretin mutation associated with cardiomyopathy and polyneuropathy. AB - We describe two Italian first cousins with familial amyloidotic polyneuropathy associated with transthyretin variant consisting of the substitution of alanine for glycine at codon 47 (TTR Ala-47), from a family with a history of cardiac failure. The 40-year-old patient presented with autonomic dysfunction and the 44 year-old cousin with congestive heart failure. Both developed sensorimotor and autonomic polyneuropathy. Since a similar clinical picture has been described in another Italian family, the cardiac involvement must be regarded as a salient and early feature of the TTR Ala-47 mutation. PMID- 10677865 TI - Single large-scale mitochondrial DNA deletion in a patient with encephalopathy, cardiomyopathy, and prominent intestinal pseudo-obstruction. AB - We studied a 62 year-old woman with a clinical phenotype characterized by encephalopathy, restrictive cardiomyopathy, and prominent intestinal pseudo obstruction. Muscle morphology showed ragged red fibres with ultrastructurally abnormal mitochondrial whereas muscle respiratory chain was normal. Molecular genetics revealed the 'common deletion' in mtDNA, which represented 40% of total mtDNA. These data expand and confirm the wide clinical spectrum of mitochondrial disorders associated with single large-scale mtDNA deletions. PMID- 10677866 TI - Identification of homozygous and heterozygous dy2J mice by PCR. AB - The dystrophia muscularis dy2J/dy2J mouse is an animal model for one form of human congenital muscular dystrophy. A point mutation in the gene coding for the laminin-2 alpha 2 chain leads to the expression of a truncated, partially functional protein. We developed a simple assay for the detection of the dy2J allele, which contains a new NdeI restriction site. Genomic DNA was prepared from animals of known status and amplified by PCR. The digestion of the PCR product with the restriction enzyme resulted in characteristic profiles. Then, this technique was used to identify heterozygous mice among unaffected animals of unknown status. Subsequently, the heterozygous genotype of these mice was confirmed by the birth of dystrophic offspring after mating. This technique allows the detection of the dy2J allele in heterozygous and homozygous animals at any age. PMID- 10677867 TI - Amyotrophic lateral sclerosis: copper/zinc superoxide dismutase (SOD1) gene mutations. AB - Mutations of the SOD1 gene, encoding the enzyme copper/zinc superoxide dismutase, have been identified in around 20% of patients with familial amyotrophic lateral sclerosis (ALS), and also in patients with apparently sporadic ALS. The table documents the mutations identified and published to date, and references clinical and pathological descriptions of the patients and families with individual mutations. The table includes 63 different mutations of SOD1 at 43 codons, three intronic sites, and two in the 3' untranslated region. Most of the mutations are heterozygotes, with autosomal dominant inheritance, but a small number of individuals appear to be sporadic, or are homozygotes with autosomal dominant recessive inheritance. PMID- 10677868 TI - 5th Workshop of the European CMT Consortium, 69th ENMC International Workshop: therapeutic approaches in CMT neuropathies and related disorders 23-25 April 1999, Soestduinen, The Netherlands. PMID- 10677869 TI - Ten years of ENMC--from a patients' initiative to a successful European research institution: the story of the European Neuromuscular Centre. PMID- 10677870 TI - Neuromuscular disorders: gene location. PMID- 10677871 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 10677872 TI - Introduction: community action research and the prevention of alcohol problems at the local level. AB - Many community action projects from around the world exist to reduce alcohol problems at the local level. The role of research within this international movement is discussed within this introduction for the entire special issue on community action research in alcohol problem prevention. Previous community prevention programs have utilized a variety of prevention strategies: (a) an educational approach which focuses on changing behavior through changes in knowledge, attitudes, and information; and (b) an environmental approach which focuses on changing behavior through changes in the social and economic systems within a community. Many projects have used both approaches. This special issue provides a current overview of many types of community action projects from different countries and summarizes what has been learned to date from these experiences. PMID- 10677873 TI - To prevent alcohol problems in Europe by community actions--various national and regional contexts. AB - The European transformation during the 1990s, concerning political, economic, social and cultural changes, has made alcohol policy in many countries very fragile and uncertain because of new social and economic conditions and decreasing power of national governments. On this background, regions, municipalities, nongovernmental organizations and the civic society are to play a more prominent role in the European alcohol policy arena. In December 1997 the first European research and evaluation symposium on community action alcohol programs was held in Malmo, Sweden. The themes were: Research-based evaluation of community action programs; The interaction between local, national, and European prevention strategies; Local programs in schools, workplaces, health sectors, primary health care, and other community contexts; The role of citizens' movements, consumer organizations, women's groups, and other voluntary organizations; Local political action. This paper is the first attempt to summarize the similarities and differences in the programs presented at the symposium. PMID- 10677874 TI - Reducing alcohol problems through community action research projects: contexts, strategies, implications, and challenges. AB - Community-based action research projects may include a number of challenges. The secular context may impede a project; for example, reducing aggregate rates of drinking-related problems may involve curtailing very popular high-risk drinking occasions. These projects may also embrace important but unrealistic goals, require matching competing goals emerging from multifoci project teams, or involve convoluted funding arrangements. Attention to team development, priority setting, and project design and evaluation issues is essential. Many projects downplay conceptual issues, such as understanding the nature of communities, organizations, systems, their operation, and social change and prevention models. Focus populations, community members and leaders, change agents team members, funding agencies, and policymakers can benefit from these projects. PMID- 10677875 TI - A decade of community action research. AB - The aim of this paper is to reflect on the past decade of research and community action on alcohol and especially on some of the presentations given in the three previous international meetings on community action: in Ontario 1989, San Diego 1992, and Greve Florence 1995. The projects reported on are diverse, reflecting the different cultures represented, but there are also common strands. Among these common strands is the growing consensus that at the heart of successful evaluated community action projects is a process of reciprocal and respectful communication: between different community sectors and also between the community and researchers. While there is increased acknowledgment of the knowledge community sectors bring to planning and implementing community action, there is also an increasing focus on the role of the researcher in providing research based knowledge to facilitate the development of effective community strategies to reduce alcohol-use-related harm. This is in contrast to a research role which emphasizes only outcome evaluation. Another development apparent through the years covered in the international meeting is the use of more naturalistic approaches to evaluation in acknowledgment that experimental design may not be feasible or scientifically appropriate for the evaluation of community action projects. PMID- 10677876 TI - Institutionalization of community action projects to reduce alcohol-use related problems: systematic facilitators. AB - This article reviews papers from a recent conference on community action research in order to identify factors that contribute to long-term maintenance, sustainability, or institutionalization of community project interventions. The descriptions of long-term outcomes and aftereffects of projects that emerged in the conference are valuable because relatively few instances of institutionalization have been documented in the scientific literature. After a general theoretical discussion of institutionalization in communities, the article identifies characteristics of successful community action programs that outlived their original funding. These characteristics include honoring community values and cultural relevance, cultivating key leader support, and utilizing indigenous staff. They also include developing local resources, maintaining flexibility, and leveraging prior success. The paper concludes by noting that aiming for policy and structural changes is a goal for an institutionalization of measures positively affecting desired health outcomes, even if the programs which created them are not themselves sustained. PMID- 10677877 TI - Community action research: who does what to whom and why? Lessons learned from local prevention efforts (international experiences). AB - This paper describes lessons learned about community action research, drawing upon papers written and presented at a recent international conference on community action research and the prevention of alcohol and other drug problems. Projects reflected both action and evaluation research traditions and focused on a variety of issues from moderation of drinking to alcohol-related violence, and on range of target populations from youth to specific ethnic groups. The interventions described ranged from policy-based prevention to education and training and to secondary prevention and treatment. Lessons identified in the papers are discussed within three broad areas: the community targeted for change; the implementation of community projects; and community action research projects generally. The common lessons emerging from these diverse projects provide useful lessons on which to base future progress in community action research. PMID- 10677878 TI - The Finnish case: community prevention in a time of rapid change in national and international trade. AB - This article looks at the present Finnish situation in planning and development of community-level prevention of alcohol and drug problems, and the experiences gained so far. Results from the first extensive evaluated project of this kind in Finland, the Lahti Project, and the program and evaluation plans for a new project in the Helsinki metropolitan area are also described. It is argued that in the present Finnish context there is need for detailed theoretical and well measured evaluation on why the results of community-based prevention are or are not achieved. PMID- 10677879 TI - Alcohol carousel and children's school drawings as part of a community educational strategy. AB - Within a community action research program, messages for the community population can be conveyed through already existing channels (newspapers, magazines, TV, radio) or special tools can be created. As part of the Rifredi Health District (16,900 inhabitants), Florence, Italy, Community Alcohol Action Research Project, 5,500 alcohol carousels (translated and adapted from the Stockholm carousel) were distributed during 1996 in the project's area where they were freely available. Two samples, one of a consumers' association (response rate 26%) and the other of school parents, employed a questionnaire. A few local key people underwent a qualitative interview. In all circumstances the carousel proved to be understandable, useful, and able to elicit discussions about alcohol issues. In 1996-97, after a 2-year training program in communication skills and alcohol prevention, 13 teachers in local preschools, elementary schools, and middle schools planned and implemented a health education program on the issues of alcohol and food. One outcome was nine drawings produced by the school children. The drawings were exhibited in some schools and supermarkets, and were hung in city buses. PMID- 10677880 TI - Community action to reduce rural drink and drive crashes in New Zealand: adapting approaches in dynamic environments. AB - This paper discusses the evolution of a two-and-a-half year pilot community action project aimed at developing strategies to reduce alcohol-use-related crashes in a rural police district in New Zealand. Formative evaluation aimed to assist an intersectoral coordinating group identify and implement strategies. Initially the idea was to establish community-based committees to mobilize on drinking and driving around the district. However, it became clear that the original concept impeded action. The focus was changed to provide more support to strengthen existing initiatives, particularly those relating to police traffic enforcement and drinking environments. This helped give the project renewed life and direction. The paper focuses on process, organizational, and external community issues affecting the project and the use of formative evaluation to assist the project to respond and adapt to dynamic circumstances. PMID- 10677881 TI - Community-identified alcohol issues in the Mexican American community: research design and utilization. AB - This article describes a community-based study, Alcohol Outlet Density and Mexican American Youth Violence, funded by the California Wellness Foundation- Violence Prevention Initiative to the prevention Research Center in Berkeley, California. The study was conducted in three northern California cities in 1993 1996. The focus is on the inclusionary planning process in designing and implementing the study. Community members were an integral part in the identification of study questions. As a result, the findings of the study are relevant to community activists in advocating alcohol-related policies. The need for more utilization-based community studies is emphasized. PMID- 10677882 TI - Time trends in alcohol habits. Results from the Kirseberg project in Malmo, Sweden. AB - This paper reports on a pilot demonstration to implement an alcohol consumption reduction project in the community of Kirseberg, located in the city of Malmo, Sweden. The objective is to present data on time trends in alcohol use habits and alcohol-use-related problems in the Kirseberg population and to discuss potential effects of the alcohol prevention activities. It was possible to implement an alcohol consumption prevention community program successfully. The analysis of time trends in alcohol use habits and alcohol-use-related problems was, however, mainly inconclusive, but indicated a decrease in alcohol consumption and alcohol use-related problems among young and middle-aged men. PMID- 10677883 TI - Malczyce, Poland: a multifaceted community action project in eastern Europe in a time of rapid economic change. AB - The major focus of this paper is the sustainability of a one-year demonstration project on drug misuse prevention, which was implemented in a local community affected by acute economic crisis and high unemployment. The project was initiated by the Institute of Psychiatry and Neurology, and supported by the European Commission. The primary goal of the project was to demonstrate that community-based prevention is possible and feasible within the context of current transitions in Poland. Its major outcome was a community prevention package consisting of a number of booklets and videos to assist other communities in their prevention efforts. Experiences from this study suggest that factors contributing to the sustainability of a community prevention project can be identified and emphasized through simple analysis of community surveys, as well as focus group discussion. PMID- 10677884 TI - Cardiovascular effects of odors. AB - Several occupational and residential settings can expose both normal and sensitive human subjects to odors and irritants. These settings include intensive agricultural operations housing swine and poultry, cigarette-smoke-filled bars, landfills and manufacturing processes. The literature suggests that adverse sensory reactions to strong odors and irritants may lead to the release of catecholamines and stress hormones. Physiological and biochemical measurements related to cardiovascular risk, e.g., blood pressure, heart rate, high-density lipoprotein (HDL) cholesterol level and serum triglyceride level, may be altered as a result of exposure to odor and irritant-induced release of catecholamines. Further work in the form of field studies and chamber exposure protocols is required to determine whether the physiological and biochemical changes observed to date represent an increase in cardiovascular risk, or are reversible changes within the normal homeostatic range. PMID- 10677885 TI - Agency for Toxic Substances and Disease Registry's 1997 priority list of hazardous substances. Latent effects--carcinogenesis, neurotoxicology, and developmental deficits in humans and animals. AB - In support of Superfund re-authorization legislation, the Division of Toxicology of the Agency for Toxic Substances and Disease Registry (ATSDR) prepared a chemical-specific consultation document for Congress that identified those chemicals with carcinogenic, neurological, or developmental adverse effects having a latency period longer than 6 years. The review was limited to the top 50 substances listed on ATSDR's 1997 Priority List of Hazardous Substances (Priority List). Among the top 50 chemicals, a review of the technical literature indicated that 38 (76%) were classified as "reasonably anticipated," "possibly," or "probably" capable of causing cancer in humans, based either on human and animal data. Eight chemicals (16%) had well-established cancer latency periods in humans of 6 years or more following exposure. Three substances (6%)--arsenic, creosote, and benzidine--had data indicating latency periods longer than 6 years. The technical literature review likewise confirmed the potential for neurological and developmental effects with a latency of 6 years. Twenty-seven (54%) of the top 50 substances caused acute and/or chronic neurotoxic effects; a number of these also caused neurological effects that persisted beyond 6 years (or the equivalent in animal studies) such as: behavioral problems, neurological deficiencies, reduced psychomotor development, cognitive deficiencies, and reduced IQ. Twenty-eight substances (56%) caused adverse developmental effects in offspring of exposed individuals or animals including increased fetal and infant mortality, decreased birth weights and litter sizes, and growth delays. Latency periods for related chemicals are expected to be similar due to structural and toxicological similarities. PMID- 10677886 TI - [Misconduct and fraud in science]. PMID- 10677887 TI - [Current aspects of handedness]. AB - Handedness is one example of many forms of behavioural lateralization seen in humans. Left-handedness has existed in a small subset of the human population, approximately 8%, since the origin of man. The incidence of left-handedness is usually reported to be consistent among human populations. Sinistrality is more common in males than in females. A vast range of testing techniques have been used to assess handedness. There are preference and performance tests. Writing hand and self-report are two of the most popular techniques. There is a strong evidence that the dominance of language functions in the cerebral cortex is different in left-handed from that in right-handed people. An understanding of handedness may provide valuables clues as to how the brain becomes organized the way it is. Several theories have been advanced over the years to explain the genesis of handedness and, in particular, left-handedness. Theories have ranged from genetic models to socio-cultural theories. Other authors suggested a pathological origin of left-handedness in man. Left-handedness runs in families and adoption studies suggests a genetic rather than an environmental origin. However, monozygotic twins appear to be substantially discordant. A polygenetic explanation which takes environmental influences into consideration is probably called for. PMID- 10677888 TI - Twenty years of the ethics committee at the medical faculty of the University of Vienna. An interim report. AB - The organisation and assessment procedures of the ethics committee of the University of Vienna medical faculty are described. Data concerning the work of the committee from 1993 through to 1997 are given in detail. Finally, the results of a survey among physicians on the acceptance of the committee's work are presented. In conclusion, the workload of an ethics committee in a large medical faculty can only be handled by efficient management of assessment procedures. However, it is difficult to achieve sufficient acceptance in a field governed by various interests. PMID- 10677889 TI - Treatment of hairy cell leukemia with cladribine (2-chlorodeoxyadenosine). AB - Hairy cell leukemia is a rare lymphoproliferative disorder resistant to conventional chemotherapeutic agents. Recently, the purine analogue cladribine (2 chlorodeoxyadenosine, 2-CdA) was introduced for the treatment of this disease. We report on 14 patients with hairy cell leukemia who were treated with 2-CdA at our department between 1993 and 1997. The patients received a single cycle of 2-CdA at a dose of 0.07 or 0.09 mg/kg/day by continuous infusion, over a seven-day period. Five patients were previously untreated, while the others had received prior treatment with interferon-alpha (seven patients), interferon-alpha and splenectomy (one patient) or interferon-alpha, splenectomy and pentostatin (one patient). Six patients achieved complete remission, three a good partial response and three partial remission. Two patients did not respond to treatment and one of them died from septicemia in aplasia. Relapse of the disease occurred in two patients. Side effects such as fever (WHO grade 2) and/or neutropenia (WHO grade 4) were noted in eight patients. Thus, 2-CdA is an effective treatment of hairy cell leukemia that can induce long lasting remissions in both, previously treated and untreated patients. PMID- 10677890 TI - The role of local anaesthetic inhalation during premedication before bronchoscopy. AB - In a prospective randomized study, the efficacy of local anaesthetic inhalation during premedication before bronchoscopic examination was evaluated. Eighty patients with chronic nonproductive cough were inhaling either nebulized anaesthetics (10 ml of 1% trimecain; 40 patients--group A) or an isotonic chlorine solution (40 patients--group B). This was followed by topical anaesthesia using spray and laryngeal syringe. Comparing the score of cough and episodes of gagging, the inhalation of local anaesthetics appeared to make the procedure slightly more comfortable for some patients. Additional anaesthesia was less frequently needed in group A than in group B (12 vs. 19 patients). However, none of the observed differences reached statistical significance. In conclusion, the inhalation of local anaesthetics at the beginning of premedication before bronchoscopy was not confirmed as a useful method that made the examination more comfortable for patients with chronic non-productive cough, but did produce a moderately beneficial effect in some of them. PMID- 10677891 TI - [Hormone replacement therapy with 17 beta-estradiol dydrogesterone: results of a 3-month open-label study]. AB - Hormone replacement therapy is well known for its beneficial effects on climacteric symptoms and is also used for the prevention of osteoporosis. In a prospective open label study we evaluated the efficacy and safety of hormone replacement therapy with 17 beta estradiol dydrogesterone (Femoston, 17 beta estradiol/continuously and dydrogesterone/sequentially). We observed 704 women who were treated with 17 beta estradiol-dydrogesterone over three months. 448 of the women previously had not used hormone replacement therapy, 224 women had been treated with a different hormone replacement therapy before they were entered into the study; for 20 women this information was not available. The physicians were asked to assess the severity of climacteric symptoms at baseline and after three months of hormone replacement therapy. In addition, the following parameters were evaluated before and at the end of the study: blood pressure, total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, blood glucose, alkaline phosphatase and gamma glutamyltransferase. Twelve women did not tolerate 17 beta estradiol-dydrogesterone and therefore dropped out of the study. Climacteric symptoms clearly improved after treatment with 17 beta estradiol dydrogesterone. During our open label prospective study, a significant decrease in blood pressure and serum levels of total cholesterol, LDL cholesterol and the LDL/HDL ratio were observed, whereas serum levels of HDL cholesterol increased significantly. Surprisingly, triglyceride levels also decreased significantly. Serum levels of alkaline phosphatase decreased significantly in women who had received a different hormone replacement therapy before they took 17 beta estradiol-dydrogesterone. We conclude that hormone replacement therapy with 17 beta estradiol-dydrogesterone is highly effective and well tolerated. Hormone replacement therapy with 17 beta estradiol-dydrogesterone appears to have a positive effect on blood pressure and the serum lipid profile. We therefore hypothesise that prolonged treatment with 17 beta estradiol-dydrogesterone may reduce morbidity and mortality secondary to cardiovascular diseases. PMID- 10677892 TI - [Austrian Society for Lung Diseases and Tuberculosis: Consensus on diagnosis and therapy of bronchial asthma in adults. Revised draft 1999]. PMID- 10677893 TI - Minimal residual disease, its detection and significance in hairy-cell leukemia. AB - As minimal residual disease (MRD) is considered the detection of hairy cells (HCs) in a patient with hairy cell leukemia (HCL) in complete remission with the absence of detectable HCs by routine morphology of peripheral blood, aspirates and bone marrow core sections, using more sensitive methods of identification as immunohistological staining or polymerase chain reaction (PCR) to detect immunoglobulin heavy chain genes rearrangement. Various monoclonal antibodies (MoAbs) as CD20, DBA.44, B ly-7, HC2, CD25 and CD11c have been applied using immunological staining. There is no standardized technique for identification of MRD. According to the technique used the MRD has been detected in 13% to 100% of patients in complete remission (CR). It may be concluded that many patients, if not all, in stable CR may have residual HCs. Whether MRD will have impact on early relapse or on long term outcome, or whether patients in CR with persistent MRD will remain so, is a matter of a longer follow-up. PMID- 10677894 TI - The influence of dimethoxybenfluron on biochemical and haematological parameters in rabbits. AB - The influence of repeated i.v. administration of dimethoxybenfluron (NO-1-B) (12 or 24 mg base/kg once weekly, 10 weeks) on biochemical and haematological parameters were studied in rabbits in vivo. No significant changes were mostly found in the serum ion levels between the dimethoxybenfluron and the control groups, as well as in most of other biochemical parameters (including total protein and albumin levels). Nevertheless, the lower dose of dimethoxybenfluron caused an increase in the glucose level. Furthermore, no significant changes were mostly present also in haematological parameters in the dimethoxybenfluron groups of rabbits (a mild decrease in thrombocytes and leucocytes). The results of our study support an assumption of good tolerance of dimethoxybenfluron from the viewpoint of its influence on biochemical and haematological parameters in rabbits and may be considered of importance for a possible therapeutic use of the derivatives. PMID- 10677895 TI - Is factor V Leiden a risk factor for fetal loss? AB - A successful pregnancy is dependent on the development of adequate placental circulation. The abnormalities of placental vasculature may result in a number of gestational pathologies, including fetal loss. The aim of our study was to determine whether women with f V Leiden are at an increased risk of pregnancy loss. For this purpose we assessed three groups of women. In a prospective group we examined 30 females with spontaneous abortions for f V Leiden. In a retrospective group we assessed the frequency of abortions in 80 women (172 pregnancies) with f V Leiden (72 heterozygous, 8 homozygous) from 57 unrelated families. In a control group we evaluated the frequency of abortions in 45 women without f V Leiden. Factor V Leiden was found in 3% of women in the 1st group. Fetal loss occurred in 10% of women in the 2nd group and in 9% in the 3rd group. Factor V Leiden was not found to be a risk factor for fetal loss in our study group. PMID- 10677896 TI - Serum soluble adhesion molecules (sICAM-1, sVCAM-1, sE-selectin) and neopterin in patients with Sjogren's syndrome. AB - Sjogren's syndrome is a systemic autoimmune disease characterized by focal lymphocytic infiltration of the salivary and lacrimal glands. Expression and up regulation of adhesion molecules and activation of cellular immune system is essential for the migration of inflammatory cells into tissues. Soluble forms of adhesion molecules sICAM-1, sVCAM-1, sE-selectin and neopterin were analyzed in serum of 17 patients with primary Sjogren's syndrome and 11 patients with secondary Sjogren's syndrome together with 26 age-matched healthy blood donors. There were significantly higher serum concentrations (mean +/- 1SD) of sICAM-1 (362.0 +/- 67.9 ng/ml, p < 0.001), sE-selectin (78.7 +/- 28.1 ng/ml, p < 0.001) and neopterin (17.9 +/- 6.4 nmol/l, p < 0.001) in primary Sjogren's syndrome patients in comparison to control group (sICAM-1: 128.3 +/- 46.9 ng/ml, sE selectin: 46.3 +/- 39.5 ng/ml, and neopterin: 7.6 +/- 2.3 nmol/l). Sera from patients with secondary Sjogren's disease contained significantly higher levels of sICAM-1 (356.0 +/- 62.4 ng/ml, p < 0.001), sE-selectin (65.5 +/- 27.0 ng/ml, p < 0.05), and neopterin (18.8 +/- 9.8 nmol/l, p < 0.001) in comparison with control group. There were no significant differences between patients with primary and secondary Sjogren's syndrome in any parameters tested. No statistically significant differences in serum levels of sVCAM-1 were found either in patients with primary or secondary SS compared to control group. PMID- 10677897 TI - Tumours and tumour-like lesions of scapula. AB - A retrospective study of 68 cases of tumours and tumour-like lesions related to the scapula, included in the registry of the Bone Tumour Reference Centre at the Institute of Pathology/University Clinics, Basle, has been carried out. Each case was evaluated for lesion entity, activity and location, age and sex of the patient, and, in 49 files with available radiographic documentation (mostly plain films), for radiologic appearance, with the aim to predict the histologic diagnosis or at least the correct dignity of the lesion. Statistically most frequent were cartilaginous tumours. More than 1/3 of all cases were osteochondromas, which demonstrated mostly a typical appearance. They were encountered predominantly in the first 3 decades in males and were located most often in the body of the scapula. 1/4 of all cases were chondrosarcomas, which were prevailing in the 4th-7th decades, but were occasionally found at a younger age too. Chondrosarcomas were located mainly at the lateral scapular margin over the inferior angle and in the acromion and coracoid process and their appearance ranged from typical to falsely benign. 1/3 of the cases represented a number of other benign and malignant histological entities. PMID- 10677898 TI - Surgery for bullous emphysema. AB - The present indications for surgery are mainly large or increasing bullae that result in compression of apparently good lung tissue, and the complications of bullous diseases such as pneumothorax. The results of local resection of localized giant bullae are dramatic. The resection of small bullae generally has little effect on lung function. Lobectomy should not be done until bullae have been removed locally and the remaining lung has been tested by positive ventilation. The indications for the resection of large bullae in the presence of diffuse emphysema require very careful individual study. Pulmonary function tests are mandatory but computed tomography is the single most useful method of assessing the extent of the bullous disease and the underlying lung disease. If the underlying lung is diffusely cystic then any surgical treatment is palliative only. PMID- 10677899 TI - [Changes in visual function in myopia 12 months after photorefractive keratectomy]. AB - 1. 110 myopes undergoing photorefractive keratectomy (PRK) with refraction from 0.25 D to -12.0 D were divided into 4 groups: A: upto -2.75 D, B: -3.0 D to -5.75 D, C: -6.0 D to -8.75 D and D: -9.0 D to -12.0 D. BCVA using the computerized method with Landolt rings and CS using an adjustment method with ascendant and descendent approach to threshold contrast adaptation on a computerized system of the Contrast sensitivity 8010 type were examined in patients before and 1 year after PRK. 20 emmetropes of the same median age were evaluated as a control group. 2. Preoperative BCVA and CS in myopes of all four groups was significantly lower (p < 0.05 az p < 0.001), compared to controls. Decrease of functions was proportional to refraction. 3. With increasing refraction less patients were within +/- 0.5 D and +/- 1.0 after surgery. 4. Twelve months after PRK, BCVA in the group A reached the level of controls, CS in the same time interval was equal to control even in groups A and B. 5. According to results of our study, PRK is a suitable method for myopia upto -6.0 D. PMID- 10677900 TI - [Hereditary occurrence of deep cavities in the optic nerve associated with Best's disease]. AB - The authors present a pedigree where in three generations in male members deep excavations of the optic discs along with Best's disease of the centres in a variable form were recorded. The heredity is autosomal dominant. Retinal changes conditioned by defects of the pigmented epithelium reduce vision to a maximum of 6/60. The anomaly is associated with a pathological EOG record and normal ERG response. The observation is a contribution to discussions in the literature about the hereditary character of excavations of the optic nerve. PMID- 10677901 TI - [Removal of silicone oils from the eyes after vitrectomy]. AB - The author describes his experience with elimination of silicone oil in 60 patients. The follow-up period was 4 to 16 months. In 30% of patients where the retina adhered, removal of the oil was associated with improved visual acuity and quality of vision. In 5%, on the other hand, removal of silicone oil was associated with reoperation of a relapse of detachment of the retina as the operation proved ineffective and the finding on the retina did not change. In 3% patients who had only one functional eye there was a subjective deterioration of vision. Before removal of silicone oil in adverse cases it is always necessary to consider carefully the risks of a relapse of detachment and deterioration of visual functions. PMID- 10677902 TI - [Clinical results in 100 corneal transplantations]. AB - The authors refer clinical results of 100 consensual penetrating keratoplasties performed at the Department of Ophthalmology, University Hospital, Hradec Kralove, in the year 1997. There were 55 women (55%) and 45 men (45%). The mean age of the patients was 58 years (range 18-84). The operation was performed for keratoconus in 24 eyes (24%), pseudophakic or aphakic endothelial failure in 18 eyes (18%), primary endothelial failure in 14 eyes (14%), traumatic changes in 18 eyes (18%), post-inflammatory changes in 7 eyes (7%). The repeated transplantation was performed in 19 cases (19%), for rejection of the previous graft in 18 eyes (94.7%) and for refractive error in 1 eye (5.3%). The average follow-up period was 10 months (range 6-17 months). For the control analysis 74 patients (74%) were available. The best corrected visual acuity was 6/6-6/18 in 21 eyes (28.4%) and 6/24-6/60 in 15 eyes (20.3%). Postoperative complications comprised rejection of the graft in 17 patients (23.0%), endophthalmitis in 3 patients (4.1%) and secondary glaucoma in 4 patients (5.4%). The results of this study indicate that despite the advantages achieved on this field, there is still scope for improvement. PMID- 10677903 TI - [A tear of the retinal pigment epithelium]. AB - Tears of the retinal pigment epithelium is less common complication of RPE ablation. There is a discussion about the pathogenesis and the reasons of the disease's origin. The presence of drusen reflects congenital changes of Bruch's membrane with a tendency to rise the RPE tears after the former detachment. Demonstrated patient before the foundation of a tear was followed up for drusen in maculae bilaterally. There is reported the very characteristical ophthalmoscopic and mainly fluorescein angiographic appearance accompanied with a sudden loss of vision and the foundation of a fibrovascular scar. PMID- 10677904 TI - [The Schmid-Fraccaro syndrome]. AB - The authors describe a female patient with bilateral colobomatous malformations of the uvea in conjunction with anorectal atresia and other symptoms suggesting Schmid-Fraccaro's syndrome called also cat eye syndrome. Using fluorescent hybridization in situ, the authors identified the supernumerous bisatellite marker chromosome derived from chromosome 22 which made it possible to confirm the suspected diagnosis. PMID- 10677905 TI - [Measurement of adhesion of the corneal lamella to the stroma in the early postoperative period after microkeratotomy]. AB - The early postoperative adhesion of corneal lamella after LASIK was measured. The interface was washed by hypo, iso and hypertonic saline solution and the influence on the lamella adhesion was assessed. Drying the interface and the excimer laser ablation of epithelial cells along the edge of the lamella were assessed in terms of the influence on the adhesion. The term "phototerapeutic epithelectomy (PTE)" was proposed for the excimer laser ablation of corneal epithelial cells. Small insignificant influence of PTE was found. The clinical application of PTE to increase the early adhesion of lamella is not recommended because of the epithelial destruction along the lamella edge. The adhesion was increased significantly by means of drying the interface which is recommended for the clinical application. High variability od measurements between different eyes was found. PMID- 10677906 TI - [Use of citation indexes and impact factors]. AB - Article deals with evaluation method of scientific works by means of Citation Indexes and Impact Factor which are produced by Institute for Scientific Information of Philadelphia. It criticizes criteria of the usage of Impact factor and way of research workers evaluation. PMID- 10677907 TI - [Surgical treatment of idiopathic macular tears]. PMID- 10677908 TI - [Giant retinal tears]. PMID- 10677909 TI - [The HAAG-STREIT AG 900BQ--a good quality and affordable endoset for corneal endothelial microscopy]. PMID- 10677910 TI - [Expression of p53, p21 and bcl-2 in prognosis of lung carcinomas]. AB - Expression of suppressor genes 53 and bcl-2 as well as of protein p21 (partly induced by p53 gene) was analyzed in a group of 77 resection specimens and bronchial excision of lung carcinomas (of all basic histological types--squamous cell, neuroendocrine, adenocarcinoma, undifferentiated). Simultaneously the relation of tumor immunophenotype and level of differentiation, cell death and 2 year-survival of patients was evaluated. Gene p53 showed non-only an expected strong expression in squamous cell carcinomas but especially in adenocarcinomas, which were newly characterized by exceptional hyper-expression of p53 in lowly differentiated variants. Expression level of protein p21 and gene p53 was parallel only in adenocarcinomas and undifferentiated carcinomas but discordant in squamous cell and neuroendocrine carcinomas. Positivity of p21 slightly prevailed in well-differentiated variants of the histological types but an exceptional positivity was found even in all the undifferentiated carcinomas. Gene bcl-2 revealed a paradox of strong expression in lowly differentiated neuroendocrine and undifferentiated carcinomas. The level of bcl-2 expression in squamous cell carcinomas was found higher than in references. Among tumors with cell death there was an inverted relation of bcl-2 and p53 expression (high/low) in neuroendocrine carcinomas but both of them were mostly negative in squamous cell carcinomas. A more frequent 2-year-survival of squamous cell carcinomas was verified for bcl-2 positive tumors and newly for p53 positive squamous cell carcinomas. Evaluation of the expression of p53, p21 and bcl-2 in lung carcinomas is so equivocal that its prognostic usage was found to be only complementary to the direct immunohistochemical investigation of the growth activities. PMID- 10677911 TI - [New diagnostic possibilities in Alport's syndrome]. AB - An overview of immunohistological and molecular genetic methods for diagnosis of Alport syndrome (AS) is given with practical experience from groups of authors' observations. Immunofluorescent investigation using antibodies against alfa chains of collagen IV was performed on cryostat sections from 29 punction nephrobiopsies and 9 skin excisions taken for support of differential diagnosis of AS particularly against the thin membranes glomerulopathy. Alfa chains deviations in other renal diseases were followed in another 14 cases. Molecular genetical investigation of AS by an indirect DNA diagnostics was performed in 35 families with presumed AS and in 27 patients with probable mutation a mutation screening of COL4AS gene by a direct method SSCP was made. The mutation was proved in 10 cases. Because of genotypical and phenotypical variability of AS the diagnostic gain only increases when all the accessible methods are combined. PMID- 10677912 TI - Ovarian hemangioma. AB - A rare case report of a clinically asymptomatic hemangioma of the left ovary is described. This tumor was an additional and accidental finding in a biopsy specimen removed because of a medium-sized mucinous cystadenoma in a 32-year-old patient. Immunohistochemically the expressions of Factor VIII and smooth muscle actin and negativity of estrogen and progesterone receptors were found. PMID- 10677913 TI - [Well differentiated inflammatory subcutaneous liposarcoma (inflammatory atypical lipomatous tumor)]. AB - Well differential inflammatory liposarcoma is a rare type of low grade liposarcoma. Its microscopical features may simulate benign inflammatory pseudotumor. A case of this lesion was described occurring in the subcutis of the right supraclavicular region in 61-year-old woman. The tumor was round well circumscribed and measured 3 cm in diameter. Histologically, it contained a 1.5 cm inflammatory nodule, which was surrounded by otherwise typical lipoma-like liposarcoma. The inflammatory nodule was composed of lymphocytic aggregates and spindle to stellate cells with mild atypism. No lipoblasts were seen in this area which mimicked an inflammatory pseudotumor. Correct diagnosis was based on the recognition of the lipoma-like liposarcoma area in this lesion. Our patient had no recurrence six years after the tumor excision. PMID- 10677914 TI - [Helicobacter heilmanii, a spiral bacterium, in gastric mucosa biopsies]. AB - Interest in possible microbiological causes of gastritis has increased significantly since the discovery of Helicobacter pylori (Hp). Recently a spiral bacterium named Helicobacter heilmannii (Hh) was described in association with chronic gastritis in adult and pediatric patients. Comparisons between these two organisms, as well as the literature on Hh, have also been reviewed. The incidence of Hh gastritis is far lower than that of Hp gastritis. Concomitant infections by Hh and Hp are very rare. It is very probable that Hh gastritis is transmitted from domestic animals or pets to humans. The frequency of Hh gastritis (11/6059 cases, 0.18%) in authors' material was similar to that reported in Western Europe. The role of touch cytology has been becoming more and more significant recently in the diagnosis of mucosal infections of the GIT. PMID- 10677915 TI - [Use of science citation indexes and impact factors]. AB - Article deals with evaluation method of scientific works by means of Citation Indexes and Impact Factor which are produced by Institute for Scientific Information of Philadelphia. It criticizes criteria of the usage of Impact factor and way of research workers evaluation. PMID- 10677916 TI - [The effect of autolysis on the interpretation of results of immunohistochemical detection of myoglobin and fibrinogen in the myocardium]. AB - Immunohistochemical detection of myoglobin and fibrinogen in the myocardium makes it possible to diagnose very early stages of ischaemic/hypoxic changes of the heart muscle. The authors demonstrate on the myocardium of a 58-years-old female patients who died suddenly with the finding of acute infarction of the anterior wall of the left ventricle with transmural rupture and cardiac tamponade the effect of autolysis on this examination. Tissue excision taken from the close vicinity of the rupture were subjected to autolysis at room temperature and immersed in formol for fixation within an interval of 24 hours. The control series of examinations was made in a similar way on the heart muscle of a 20 years-old woman who died from violent death, and ischaemic changes of the heart muscle were not anticipated. While in the early stages of autolysis it was possible to use combined immunohistochemical detection of myoglobin and fibrinogen, after a longer postmortal interval extensive artificial losses of myoglobin were observed, and the method of detection of fibrinogen deposits in damaged cardiomyocytes seemed more suitable. Even after a postmortal interval of 168 hours it was possible to differentiate reliably damaged myocytes with fibrinogen deposits from intact muscle fibres, where fibrinogen deposits were not observed. PMID- 10677917 TI - [Overview of the changes in alcohol content of beers produced in the Czech Republic]. AB - Paper concerns the alcohol contents of beer now produced. The previous and contemporary ways of beer signing as well as a complete list of beer sorts produced in the Czech Republic is presented. PMID- 10677918 TI - Impact of hepatitis B and C virus infection on the outcome of kidney transplantation in Chinese patients. AB - BACKGROUND: There has been improvement in kidney transplantation over the years; however, the impact of hepatitis B and C virus (HBV, HCV) infection on the long term outcome of kidney transplant is still controversial. METHODS: A total of 113 patients who received renal allografts from 1986 to 1998 were analyzed. Nine were positive for both hepatitis B surface antigen (HBsAg) and antibody to HCV (anti HCV) (Group 1), 20 were HBsAg-positive and anti-HCV-negative (Group 2), 30 were HBsAg-negative and anti-HCV-positive (Group 3) and 54 were negative for both markers (Group 4). The outcome and survival were compared among the four groups of patients. RESULTS: The mean follow-up period was 5.1 +/- 3.2 years (range, 0.5 13 years) for all patients. Group 2 patients had significantly higher liver related complications (35% vs 0%, p < 0.0001) and liver-related deaths (20% vs 0%, p = 0.004) than did Group 4 patients. Among all, four HBsAg-positive patients had fulminant hepatitis and died within two years of transplantation. Three (Group 2) of the patients who died were seropositive for hepatitis B e antigen and/or HBV DNA and none had a history of or positive serologic marker to indicate hepatitis of other etiologies. The remaining patient (Group 1) had evidence of superinfection of HCV. Liver cirrhosis occurred in one, two and one patient in Groups 1, 2 and 3, respectively, and hepatocellular carcinoma occurred in two and one patient in Groups 2 and 3, respectively. Despite high liver-related mortality in HBV-infected patients, paradoxically, no significant differences among the four groups in the long-term graft and patient survivals were demonstrated. The presence of HBsAg or anti-HCV was not associated with a poor prognosis of survival as determined by Cox regression analysis. CONCLUSIONS: HBV or HCV infection is not a contraindication to kidney transplantation in Chinese patients. However, it should be noted that serious liver-related complications may occur and limit survival in HBV- and/or HCV-infected patients after kidney transplantation. PMID- 10677919 TI - Significantly higher levels of oxidized LDL autoantibody in coronary artery disease patients. AB - BACKGROUND: Increasing evidence shows that oxidized low-density lipoprotein (ox LDL) might play an important role in the pathogenesis of atherosclerosis. Ox-LDL is immunogenic and induces an autoantibody, which we used as a tool for measuring the content of ox-LDL in vivo. METHODS: Patients who were admitted for diagnostic cardiac catheterization for typical or atypical angina pectoris were enrolled in this study. After fasting for 12 hours, a venous blood sample was drawn from the antecubital vein for testing triglyceride, total cholesterol, LDL cholesterol, high-density lipoprotein (HDL) cholesterol, and ox-LDL autoantibody. The ox-LDL autoantibody was quantified using an enzyme linked immunosorbent assay. All patients underwent coronary angiography. Those who had more than 50% angiographic coronary luminal stenosis, were grouped into the coronary artery disease (CAD) group. RESULTS: Sixty-four patients were enrolled in the study (male/female = 46/18; mean +/- standard deviation, age, 64 +/- 9 years). The CAD group had a significantly higher level of ox-LDL autoantibody than the non-CAD group (494.0 +/- 355.0 mU/ml vs 258.1 +/- 196.8 mU/ml, p = 0.004). However, the other lipid profiles including triglyceride, total cholesterol, LDL-cholesterol and HDL cholesterol were not statistically different between the two groups. Forty-six patients in this study had an arterial blood sample taken from the femoral artery for testing ox-LDL autoantibody. There was no significant difference between the arterial and venous samples of ox-LDL autoantibody (385.2 +/- 333.3 mU/ml vs 399.3 +/- 339.5 mU/ml, n = 46, p = 0.530). CONCLUSIONS: Ox-LDL autoantibody was significantly higher in the CAD group. Ox-LDL may prove to play a key role in the pathogenesis of atherosclerosis. Further study of Ox-LDL and its role in the process of atherosclerosis is warranted. PMID- 10677920 TI - Catheter-induced coronary spasm--a view of mechanical factors and experience with selective left coronary arteriography. AB - BACKGROUND: Coronary spasm during cardiac catheterization is not unusual. The mechanism of spasm remains uncertain, but is considered to be multifactorial. Many researchers believe that coronary spasm that develops during catheterization is partly spontaneous and partly catheter-induced. Because catheter-induced spasm results from mechanical irritation, we tried to find the iatrogenic factors that predispose patients to coronary spasm during coronary angiography. METHODS: Retrospectively, we reviewed the records of 7,295 patients who underwent coronary angiography at our hospital from June, 1983 to November, 1997; coronary spasm was documented in 30 patients, who became the study group. We randomly selected 41 patients who had normal coronary arteries as the normal control group. After reviewing cine films of coronary angiography, we compared these two groups for several parameters. These parameters included the length and diameter of the left main coronary artery (LMC), the angle between the LMC and the aorta, the angle between the catheter tip and the LMC, whether the catheter tip came into contact with the vascular wall and whether there was vessel wall bulging, catheter size and catheter/LMC ratio. This angiographic data and the demographic features, including age, sex, history of hypertension, diabetes mellitus, smoking, previous myocardial infarction, family history of coronary artery disease, cholesterol and triglyceride levels and chest pain character (exertional or rest pain) were compared between the study patient group and the control group. RESULTS: The results disclosed that larger catheter size (7.1 +/- 0.6 mm vs 6.4 +/- 0.7 mm, p < 0.001), smaller LMC diameter (4.2 +/- 0.9 mm vs 4.9 +/- 1.0 mm, p = 0.004), larger catheter/LMC ratio (0.07 +/- 0.05 vs 0.05 +/- 0.03, p = 0.022), catheter contact with the vessel wall (27/30 vs 20/41, p < 0.001) and vessel bulging (18/30 vs 5/41, p < 0.001) were related to catheter-induced coronary spasm. We found that the catheter tip coming into contact with the vessel wall, vessel wall bulging and catheter/LMC ratio (odds ratio 8.92 x 10(14)) were statistically significant factors predisposing patients to catheter-induced coronary spasm. CONCLUSIONS: Multiple factors contribute to coronary spasm. Of those, mechanical or iatrogenic factors might predispose patients to spasm during coronary catheterization. These facts deserve our attention, because iatrogenically induced spasms may be avoided by meticulously selecting catheters and manipulating them gently. PMID- 10677921 TI - Influence of morning or evening administration on absorption of theophylline. AB - BACKGROUND: Bronchoconstriction during the night causing nocturnal and early morning wheezing is recognized as a major problem for asthmatics. Oral sustained release theophyllines (SRTs) were developed to reduce the symptoms. A circadian variation in theophylline kinetics has been demonstrated with many SRTs. The purpose of this study was to evaluate the differences in serum theophylline concentration (STC) caused by morning or evening dosing of Euphyllin Retard, a brand of SRT, for a period of 36 hours following oral administration. METHODS: A total of nine non-smoking healthy male volunteers were involved in the study, with a two-period crossover comparison. They were randomly divided into two groups. The first group took a single oral dose of 350 mg Euphyllin Retard at 8:00 A.M. and the second group took it at 8:00 P.M. Blood samples were collected during the 36 hours following administration. Two weeks later, the first group took the drug at night and the second group took it in the morning. The difference in the absorption of theophylline with daytime administration versus night-time administration was assessed using pharmacokinetic parameters derived from the plasma drug concentration vs time curve. RESULTS: The means of unextrapolated area under the concentration vs time curve (AUC) from time 0 to 24 hours (AUCUN) and of the extrapolated AUC from time 0 to infinity (AUCEX) in the night phase were higher than those in the day phase (62.403 micrograms/ml/hr vs 53.081 micrograms/ml/hr, p = 0.9186; 107.21 micrograms/ml/hr vs 98.879 micrograms/ml/hr, p = 0.8807, respectively). The mean of maximum concentration (Cmax) was higher in the night phase than that in the day phase (4.166 micrograms/dl vs 3.451 micrograms/dl, p = 0.9234). Daytime administration showed a delayed time to maximum concentration (Tmax) when compared to that of night time administration (6.5 hr vs 5.75 hr, p = 0.6244). The terminal elimination rate constant (Kel) was lower in the day phase than in the night phase (0.053 l/hr vs 0.06 l/hr, p = 0.7601). The day phase and night phase data are combined data from the two night and two day groups. The statistical analysis of the results show that the time of administration does not influence the STC. CONCLUSIONS: No diurnal variation in theophylline kinetics was found with Euphyllin Retard. This study was performed in a limited number of normal healthy subjects, and the same result is yet to be proved in asthmatic patients and a larger population of normal subjects. PMID- 10677922 TI - In vitro activity of quinupristin/dalfopristin and other antibiotics against ampicillin-resistant enterococcus faecium. AB - BACKGROUND: Enterococcus faecium constitutes approximately 10% of clinical isolates of enterococci and is noted for its antimicrobial resistance. In particular, E faecium is commonly resistant to ampicillin. The optimal treatment for severe infections caused by these multi-resistant organisms has yet to be determined. METHODS: Enterococci tested were isolated from blood, pleural fluid and cerebrospinal fluid. Ampicillin-resistant Enterococcus faecium (AREF) was identified using the API Rapid Strep Kit system. A total of 58 isolates of AREF were enrolled in this study. Ten different antibiotics were tested, including Synercid (quinupristin/dalfopristin), teicoplanin, vancomycin, ampicillin, trimethoprim/sulfamethoxazole (TMP/SMX), ciprofloxacin, gentamicin, chloramphenicol, rifampicin and tetracycline. The agar dilution method described by the National Committee for Clinical Laboratory Standards was used to determine the minimum inhibitory concentrations (MICs) of the antibiotics tested. RESULTS: Teicoplanin showed the best in vitro activity. Its MIC ranged from 0.25 to 2 micrograms/ml with an MIC90 of 1 microgram/ml. The MIC of vancomycin was 0.5-128 micrograms/ml with an MIC90 of 2 micrograms/ml. Three strains were vancomycin resistant, and they were the VanB phenotype. The MIC of quinupristin/dalfopristin was 0.5 to 8 micrograms/ml with an MIC90 of 2 micrograms/ml. Chloramphenicol and tetracycline showed moderate susceptibility. AREF showed high resistance to other antibiotics tested, including ciprofloxacin, gentamicin, TMP/SMX and rifampicin. High-level gentamicin resistance (MIC > 1,000 micrograms/ml) was found in 78% of AREF tested. CONCLUSIONS: Teicoplanin showed the best in vitro activity against AREF. Clinical studies are necessary to confirm the efficacy of quinupristin/dalfopristin in vivo. PMID- 10677924 TI - Lymph node revealing solution and traditional 10% buffered formaldehyde for detecting lymph nodes in colorectal carcinoma. AB - BACKGROUND: Cancer stage has been the most important factor in evaluating the prognosis and treatment of colorectal carcinoma, and the presence of lymph node metastasis in the resected specimen plays a key role in serial staging. We evaluated a newly proposed "lymph node revealing solution" (LNRS) that proponents claim is inexpensive, easy, rapid, and innocuous, and that could help to establish the stage of disease more accurately than buffered formaldehyde (formalin). Our study compared traditional 10% formalin and LNRS as second search solutions to determine whether LNRS is a more useful and practical solution than formalin. METHODS: Thirty randomly selected colorectal carcinomas from low anterior resection (LAR) were studied. The specimens were handled routinely on the first day. Then, the entire fat of odd-numbered cases was immersed in three times its volume of LNRS, and the entire fat of even-numbered cases was immersed in 10 times its volume of formalin overnight. The chi-squared test was used to evaluate the results. RESULTS: Of 145 cassettes in the second search by LNRS, seven had no lymph nodes. Of 107 cassettes in the second search by formalin, 18 had no lymph nodes. Totally, 792 lymph nodes were found with an average number of 26.4 per case and 1.53/cm (per centimeter of specimen) in all 30 cases. By the LNRS method, 138 lymph nodes were found in the 15 odd-numbered cases, and by the formalin method, 89 lymph nodes were found in the 15 even-numbered cases. We were able to adjust the stage upward in two cases by LNRS-fixation method. There were no changes in staging in the formalin-fixation method cases. CONCLUSIONS: LNRS showed no statistically significant difference from formalin in the numbers of total lymph nodes, positive lymph nodes, or minute (less than 1 mm) lymph nodes found. Nonetheless, we do believe that LNRS is a potential replacement solution for formalin due to its fast fixation, ease of searching, high identification rate and time-saving procedure. PMID- 10677923 TI - Prediction of biochemical relapse for stage pT3 prostate cancer following radical prostatectomy. AB - BACKGROUND: Approximately 30% to 50% of clinically localized prostate cancers are found to be locally advanced after radical prostatectomy. Without adjuvant therapy, more than 30% of stage pT3 patients will have biochemical failure in a median follow-up of three years. Whether adjuvant therapy should be given remains controversial. Identification of prognostic indicators may be helpful to select patients at risk of biochemical failure for postoperative adjuvant therapy. METHODS: The medical records of 22 pT3 prostate cancer patients were analyzed retrospectively. Postoperative management included surveillance (7 patients), adjuvant hormone therapy (7) and radiotherapy (8). Patients were monitored every three months using serum prostate-specific antigen (PSA). Biochemical failure was defined as an elevation in PSA of more than 0.2 ng/ml. The clinical outcome was correlated with tumor grading, pathologic staging, preoperative PSA level, and p53 and bcl-2 status. RESULTS: Five patients (23%) experienced biochemical relapse within a median follow-up time of 52 months (range, 10-71 months). A Gleason score of 8 or more (p = 0.001) and seminal vesicle involvement (p = 0.014) were significant prognostic indicators in the univariate analysis. Patients with low preoperative PSA levels (< 20 ng/ml) had a significantly higher failure rate (p = 0.031). However, most of these patients (56%) had tumors with Gleason scores of 8 or more. Neither p53 gene mutation nor bcl-2 overexpression predicted PSA failure. Postoperative adjuvant therapy did not appear to reduce the risk of disease recurrence. In multivariate analysis, the Gleason score was the only significant factor predicting biochemical failure (p = 0.017). CONCLUSIONS: Patients with poorly differentiated tumors and/or seminal vesicle invasiveness are at higher risk of biochemical failure after radical prostatectomy. Our limited experience did not support the routine application of adjuvant therapy for this subgroup of patients. A larger sample size with a longer follow-up period is necessary to reach a definitive conclusion. PMID- 10677925 TI - Midbrain hemorrhage presenting with trochlear nerve palsy. AB - A 40-year-old normotensive man suddenly developed diplopia, tinnitus and a burning sensation on the left side of his body while driving a motorcycle. He did not complain of headache, nausea or vomiting. Neurologic examination revealed left trochlear nerve palsy and impaired pinprick, temperature and joint position sensation of the left limbs. There was no ptosis or motor deficit. He had a mild bleeding diathesis due to alcoholic liver cirrhosis. Computerized tomography and magnetic resonance image of the brain disclosed hemorrhages in the right midbrain tectum and the left temporal lobe. After nine months of observation, there was nearly complete recovery of symptoms, except for mild residual diplopia. From a literature review, only nine case of midbrain tectal hemorrhage involving the inferior colliculus have been reported. These patients had a unique clinical presentation. Diplopia due to trochlear nerve palsy, either unilateral or bilateral, was present in all of the cases. Tinnitus and sensory disturbance contralateral to the lesion side were very common. Only three patients had risk factors for hemorrhage, including bleeding diathesis, hypertension and vascular anomalies. In the majority of patients, no underlying causes were detected. The outcome was favorable with conservative treatment. PMID- 10677926 TI - Symptomatic esophageal stricture after endoscopic variceal ligation--success of endoscopic balloon dilation. AB - Endoscopic variceal ligation is a preferred endoscopic method to treat esophageal variceal bleeding. Esophageal stricture complicated by endoscopic ligation is very unusual. We report a case of symptomatic esophageal stricture after variceal ligation. Endoscopic balloon dilatation was used to treat the stricture successfully. The possible mechanism of stricture formation is discussed. PMID- 10677927 TI - Immediate repair of intestinal injury during laparoscopically assisted vaginal hysterectomy. AB - With the increasing popularity of laparoscopic surgery in gynecologic diseases, complications with this procedure are noted more frequently. The majority of complications occur during operation, and most of the cases are immediately identified. The recovery and outcome are relatively good due to urgent repair and management. Herein, we report a complication--a trocar, inducing small intestinal damage, which was detected during laparoscopically assisted vaginal hysterectomy. The damage was repaired promptly extraperitoneally with an uneventful recovery. Through a review of the literature and our experience, possible preventive methods of this complication are discussed. In addition, by this case review, we hoped that such a complication could be avoided in the future. PMID- 10677928 TI - Coexistence of epilepsy, myasthenia gravis and psoriasis vulgaris. AB - We report the case of a 36-year-old Chinese man with a history of complex partial seizure of temporal lobe origin since the age of 12 years, superimposed by myasthenia gravis since the age of 27 years and psoriasis vulgaris since the age of 29 years. With an eight-year follow-up, the above three diseases remained without complete remission. Anticonvulsant therapy (phenytoin and trimethadione) caused drug-induced myasthenia gravis, which should gradually disappear after discontinuing the drugs. However, the myasthenic symptoms and serum acetylcholine receptor antibody persisted following the discontinuation of phenytoin in our patient. Myasthenia gravis and psoriasis are both autoimmune diseases and correlate with specific human histocompatibility antigens. This suggests a close connection between these two diseases. The coexistence of epilepsy, myasthenia gravis and psoriasis vulgaris has not been previously reported, and to the best of our knowledge, our patient is the first reported case. The relationship among these three diseases requires further investigation. PMID- 10677929 TI - Endoscopic sinus surgery for ethmoid sinus meningioma. AB - Meningiomas in the ethmoid sinuses are a challenge to manage. A 50-year-old man suffered from a left olfactory groove meningioma. He underwent a bilateral craniotomy to remove the tumor mass in August, 1997. During the follow-up period, a tumor was found in the right posterior ethmoid sinus. Endoscopic sinus surgery was performed to remove the tumor mass in August, 1998. Pathologic examination of the mass revealed a meningioma. No intraoperative or postoperative complications occurred, except for an episode of seizure. PMID- 10677930 TI - Successful birth after intracytoplasmic sperm injection for severe male factor infertility in a woman with poor response to controlled ovarian hyperstimulation. AB - Poor responders to controlled ovarian hyperstimulation (COH) present a clinical challenge for in vitro fertilization (IVF) and embryo transfer. The failure of IVF for the treatment of severe male-factor infertility can now be overcome by intracytoplasmic sperm injection (ICSI). The infertile couple documented in this case report came to our hospital because of bilateral tubal occlusion and severe oligoasthenospermia. After three poor-response cycles to COH, one mature oocyte was retrieved and was fertilized using ICSI. Normal fertilization ensued and one good-quality, eight-celled embryo was transferred into the woman's uterus. A single gestation was confirmed by ultrasound seven weeks after transfer. Amniocentesis was performed at 16 weeks and demonstrated a normal male fetus with a karyotype of 46,XY. The patient had a spontaneous, normal, vaginal delivery of a 2,650 g healthy male infant. PMID- 10677931 TI - Congenital muscular dystrophy. AB - Congenital muscular dystrophy (CMD) is a rare heterogeneous disease found in the oriental population, especially the occidental type of CMD. We report a case of a one-year-old infant who presented with early onset hypotonia, muscular weakness, delayed motor development and normal intelligence. A muscle biopsy revealed dystrophic muscle fibers. A high creatine kinase (CK) level, mostly of the MM type, was also noted. Further study of brain images showed hyperintense lesions in the white matter area. The patient showed the clinical and laboratory findings characteristic of CMD, more likely to be of the occidental type. Further genetic or histopathologic studies, especially merosin investigation, are suggested for improved classification and prognosis prediction. PMID- 10677932 TI - Chronic myeloid leukemia initially presenting with spontaneous mediastinal hematoma and hemothorax. AB - Spontaneous mediastinal hematoma is rarely seen in hematologic malignancy. We report a case of chronic myeloid leukemia initially presenting with spontaneous hematoma and hemothorax. In addition to a detailed history, computerized tomography of the chest is important in analyzing whether an anterior mediastinal mass lesion is present. Magnetic resonance imaging is helpful in confirming the nature of a mediastinal hematoma. Trauma, vascular disease and coagulopathy should first be ruled out when making a diagnosis of spontaneous bleeding in the thorax. In our patient, the mediastinal hematoma regressed spontaneously after three months. Leukemia should be considered in the differential diagnosis of spontaneous mediastinal hematoma. In leukemia patients with spontaneous mediastinal hematoma, supportive observation and close follow-up may be better than surgery, unless massive hemorrhage or active bleeding in the thorax is suspected. PMID- 10677933 TI - [nm23 expression and its correlation with lung metastasis in human salivary adenoid cystic carcinoma]. AB - The nm23 gene products, nucleoside diphosphate kinase (NDPK), expression in salivary adenoid cystic carcinoma (ACC) was evaluted by using LSAB immunohistochemical method. Of 25 cases tested, 16 (64.0%) showed positive staining, in which, higher incidence of positive staining was found in ACC without lung metastasis (88.2%, 15/17) than in that with lung metastasis (12.5%, 1/8; P < 0.01). Expression of NDPK/nm23 was correlated with the P-TNM pattern (P < 0.05), otherwise it was not correlated with pathologic types (P > 0.05). The result suggest that the nm23/NDPK may play a role in suppressing the metastatic potential in ACC. PMID- 10677934 TI - [Serotype distribution of Actinobacillus actinomycetemcomitans]. AB - A total of 131 Actinobacillus actinomycetemcomitans clinical strains from 28 periodontally healthy or diseased subjects were serologically categorized by indirect enzyme-linked immnosorbent assay. Sepecies-specific and serotype b, c specific monoclonal antibodies against Actinobacillus actinomycetemcomitans were used. Preliminary data revealed that each subject harbored only one serotype of Actinobacillus actinomycetemcomitans. The distribution of serotype a, b and c were respectively 11%, 21% and 68%, both of periodontally healthy subjects harbered serotype b, 7 of 9 juvenile periodontitis subjects harbered serotype c, 12 of the other subjects which included 3 adult periodontitis and 14 gingivitis harbered serotype c. PMID- 10677935 TI - [Surgical correction of dentofacial deformities following temporomandibular joint ankylosis]. AB - The paper presents 22 patients (9 male, 13 female; 13 unilateral, 9 bilateral) with temporomandibular ankylosis and dentofacial deformities treated by arthroplasty and orthognathic operations. The results were satisfactory. The key point is to ensure the function of mouth opening and correction of dentofacial deformities caused by ankylosis. The indications for one-stage or two-stage surgical correction are discussed. PMID- 10677936 TI - [Mast cells in the labial cancer: histochemical and electron microscopical study]. AB - The labial cancer tissue of 24 cases and normal labial tissue of 7 cases were studied histochemically and electron microscopically. The result showed that the number of mast cells around the periphery of labial cancer tissue increased significantly. The histochemical properties of these mast cells differed from those in the normal labial tissue. The former did not contain heparin which was present normally. Under electron microscope, according to the characteristic ultrastructure, the mast cells around the labial cancer were TC mast cells. Some of them showed the appearance of degranulation, and contacted intimately with cancer cells, lymphocytes and macrophages, etc. These findings suggest that together with lymphocytes, macrophages, these mast cells contribute to the defence reaction to the cancer tissue. PMID- 10677937 TI - [Experimental study on bone formation in a denser coral used for repairing cortical defects in dogs]. AB - The present study was designed to investigate a coral skeleton, Favites, which was originated from the reef builders in Hainan island, as a bone substitute. The coral Favites, like porites, is composed of anagonite (CaCO3), but contains less volume of porosity, higher strength and denser construction than Porites. After implanted into cortical bone defects in mandibles and femurs of dogs, skeleton fragment of coral Favites demonstrated good biocompatibility and biodegradability. A resorption of the implant and a simultaneous apposition of new bone was observed. Coral Favites was totally replaced by newly formed bone after a period of time from 6-8 months. Compared to Porites, Favites has a lower rate of resorption and therefore the defects of host bone are repaired completely without any lost of bone volume. The results showed that coral Favites may be used as a bone substitute for the segmental defects of mandibles. PMID- 10677938 TI - [Finite element analysis of force produced by "T" loop retraction archwire]. AB - This study introduced three presumed springs to imitate the three dimensional bolstering function of the bracket-tooth-periodontal tissue on the archwire with three dimensional finite element method (FEM), and, systematically analysed the force system produced by the "T" loop retraction archwire on the incisors. The following conclusions were drawn; (1) The activation of the "T" loop retraction archwire will produce extrusion and root lingual torque force as well as horizontal force on the incisors; (2) Adequate torqul on the incisor segment and gable bend mesial to the T-loop are necessary to control the position of the anterior teeth while they are being retracted; (3) retraction archwire should not be activated too much; (4) Lighter wire will produce milder and more durable forces. PMID- 10677939 TI - [Clinical studies on pulpitis and periapical periodontitis caused by traumatic occlusion]. AB - 89 cases including 100 teeth with pulpitis and apical periodontitis caused by traumatic occlusion were clinically observed. All teeth are caries-free, no any dental disease, and no pocket formation. But there is a distinct evidence of marked occlusal trauma, such as premature contact and occlusal interference. The single-root teeth are usually involved. When subjected to vitality testing the teeth associated with the periapical lesions sometimes yield positive response (about 30%). The result of investigation endodontic treatment demonstrated that the presence of predisposing factors of trauma from occlusion led to decrease of endodontic success. PMID- 10677940 TI - [Electron microscopy of elastic system fibers in the rat mandibular joint]. AB - We study the Elastic System Fibers (ESFs) in the rat mandibular joint using light microscope and image analyzer system and Electron Microscope. The results showed that the ESFs in the articular disc and capsule were more than in the articular cartilage. In addition, microfibrils and elaunin were the principal ESFs in all the articular components. The microfibrils in the articular surfaces ran at nearly right angle to the collagen fibers. The ESFs beneath the articular surfaces and the other articular components showed no specific directivity. ESFs construct an extensive fibers network in the disc and the capsule. PMID- 10677941 TI - [Using Nd-YAG laser to desensitize the hypersensitive dentine in preparing the abutment teeth for removable partial dentures prosthesis]. AB - Patients often suffer from sensitivity and pain while preparing the seats of occlusal rests of clasps on abutment teeth with hypersensitive dentine for removable partial dentures prosthesis. Base Nd-YAG laser desensitization effect by making a hard surface layer in the hypersensitive dentine of the abutment teeth, We use this method to treat the teeth before and after occlusal seats of clasps preparation, 20 patients, more than 30 occlusal seats of clasp rests are completed smoothly without any suffering since 1991. All the patients wear their dentures satisfactorily without afterward side effect. After half one year to four years followed observation all the rests adapt to their seats well without marking wear, We believe that the desensitizing effect of Nd-YAG Laser is effective in preparing occlusal seats for removable partial denture prosthesis. PMID- 10677942 TI - [Quantitative histological study of adult condylar proliferation zone]. AB - The thicknesses and cell numbers of 43 adult condylar proliferation zone were observed by serial section and multiarea measurement in order to investigate the relationships between them and age. The samples were divided into three groups by age (19-45, 46-59, 60-74). By way of statistical analysis, the cell numbers and thicknesses of different areas of the same age group, different age of the same area, and the relationship between thicknesses and age was analysed. The results show that the average proliferation zone thickness is different among different age groups, it becomes thinner with the increase of age, and the cell number is decreased. There is high significant difference among different age groups (P < 0.01). But there is no significant difference of the proliferation zone thickness between the group of 45-59 years old and the older group (60-74)(P > 0.05). There is diverse correlation between thicknesses and age increase. PMID- 10677943 TI - [A bio-electrochemical theory in pathogenesis of dental caries]. AB - Since the Bio-electrochemical theory in pathogenesis of dental caries was suggested by Huang (1990), many experimental proofs have obtained in studies of dental bio-electrical potential and dielectric properties of the tooth. Recently, the model of the electrochemical artificial dental caries that have directly proved Huang's theory was developed successfully in laboratory. This paper make a systematic exposition of the bio-electrochemical theory in pathogenesis of dental caries. PMID- 10677944 TI - [Mandibular ridge augmentation by sandwich osteotomy and BMP-HA implantation]. AB - In order to solve a series of problems ridge augmentation by subperiosteal implantstion, HA combined with BMP was implanted in between the two pieces of mandible produced by osteotomy. Nine adult mongrel dogs were used in this study and six patients were treated with this mathod. The specimens were examined by light microscope, scanning electron microscope in the experimental study. The results show that there are a large amount of new bone formation both within and around the combined material. The amount is increasing as the time lapsed and the new bone is connected directly with the host bone of two side. Clinical application showed that after augmentation of the atrophic alveolar ridge with this mathod, the shape and volume of the alveolar ridge were good. The retention and stability of the lower denture was improved significantly. The results showed that this method of alveolar ridge augmentation has many advantages. PMID- 10677945 TI - [The experimental study about the relations between Chinese herb-epimedium leptorrhizum stearn (CH-ELS) and endogenous cAMP in alveolar bone of orthodontic tooth in rats]. AB - The aim of this study was, to explore the mechanism of CH-ELS promoting bone remodelling of orthodontic tooth in rats. 11 male sprague-Dawleg rats was divided into 3 groups. Group I (normal control group) Group II (applied force, no injection) Group III (applied force, injection). The experimental period was 3 days. The radioimmunoassay (RIA) method were used to investigate the effects of CH-ELS on the content of endogenous cAMP in alveolar bone of orthodontic tooth in rats. Results indicate that CH-ELS has no significant effects on endogenous cAMP in alveolar bone of orthodontic tooth, perhaps, cAMP were not involved to regulate the mechanism of CH-ELS promoting bone remodelling. To explore the mechanism of bone remodelling of orthodontic tooth is more significant in orthodontic clinic. PMID- 10677946 TI - [Significance of glycosaminoglycans content in developing mechanism of cleft palate]. AB - To explore significance of Glycosaminoglycans content in the development of embryo and forming mechanism of cleft palate, Dexamethason acetate was injected into the abdomen of NIH mouse on the 12th day of gestation to induce cleft palate mouse embryo model. Then by taking cranio-maxillary coronary sections, stained with Alcian blue/PAS and quantitatively analysing the area of the palatal process covered by glycosaminoglycans using a TDS-90 photodigital system with computer. Results indicate that on 13(12) day(hour) of gestation the colour of Alcian blue starts to show in the palatal process. By then, the area covered by glycosaminoglycans in the normal group is 4.69 mm whereas for the experimental group is 2.40 mm, showing a significant statistical difference (P < 0.05). The area of the palatal process covered by glycosaminoglycans gradually increases as the embryo developes, but is distinctly smaller in experimental than in the normal group (P > 0.05). This proves that there's a relationship between the level of glycosaminoglycans in the palatal process and development of cleft palates. PMID- 10677947 TI - [Effects of IL-1 on experimental tooth movement in rabbits]. AB - The purpose of this study was to search the effects of IL-1 on the orthodontic tooth movement and alveolar bone by animal experiment. The distance of tooth movement was measured and histologic changes were observed. 12 rabbits were divided into two groups. IL-1 was injected in the gingiva around the moving tooth of the experimental group. The rate of tooth movement and the number of osteoclasts of the two groups were compared. The results suggested that IL-1 can promote the tooth movement and increase the number of osteoclasts and can also promote bone remodelling. PMID- 10677948 TI - [The alveolar bone density in rats with experimental diabetes]. AB - The alveolar bone turnover in rat with experimental diabetes was investigated. The serum osteocalcin concentration in diabetic rat was shown to increase as determined by radioimmunoassay; while alveolar and femur bone density decrease as determined by Dual Energy X-ray Bone Densitometer; histological examination showed that bone resorption was increased and bone formation decreased in the alveolar bone in diabetic rat. It is suggested that the alveolar bone in rats with diabetes mellitus have a tendency to develop osteoporosis. PMID- 10677949 TI - [Effects of overdose fluoride on metabolism of extracellular matrix collagen and proteoglycan in condylar cartilage]. AB - The study was conducted to evaluate the effects of chronic fluorosis on the metabolism of extracellular matrix in the development of rabbit condylar cartilage using the technique of pathology, biochemistry and radiographs. The results demonstrated that collagen content of condylar cartilage and the bone under condylar cartilage decreased (P < 0.01), and the proteoglycar contents increased significantly during the process of fluorosis (P < 0.01). The chondroprogenitor was obviously thin, and the cells of maturation zone increased in number. All the results indicated that overdose of fluoride retarded the growth of the condyle and ramus of the mandible. PMID- 10677950 TI - [The behaviour and treatment of myoepithelial carcinoma of salivary glands]. AB - Nineteen cases of myoepithelial carcinoma (malignant myoepithelioma) in salivary glands were studied clinicopathologically. The clinical features are as follows: arise most frequently within the parotid glands, followed by the submandibular glands; rapidly enlarging mass with extensive invasion of the surrounding tissues in some cases; low rate of cervical lymph node metastasis but high rate of distant metastasis; very frequent recurrence after surgical excision; and poor prognosis. The carcinoma belongs to the high grade malignant tumor. It must be treated radically. The elective neck dissection is generally unnecessary. It is resistant to radiotherapy. It is possible to get rather good result in limited recurrent tumors by prompt surgery. PMID- 10677951 TI - [Detection of types I, III and IV collagen in human cementum, periodontal ligament and alveolar bone]. AB - The aim of the present study was to investigate distribution and composition of collagen Types I, III and IV in human cementum, periodontal ligament and alveolar bone with immunohistochemical localization and image analysis technique. The results showed that collagen Type I was 78.06% in periodontal ligament, 73.09% in cementum and 30.50% in alveolar bone; Type III was 11.73% in periodontal ligament and was also observed in apical cementum and bone tissue of alveolar socket; Type IV was restricted to the basement membrane of the blood vessles; Sharpey's fibers was stained by Type I and its periphery was demonstrated stronger staining by Type I and Type III as a sheath. These results provided necessary informations on healthy periodontal tissues that will be required for future studies on the effects of pathological and regenerative processes. PMID- 10677952 TI - [Titanium plate reconstruction and fixation system for maxillofacial surgery: a synthetic clinical and experimental investigation]. AB - Titanium plate-screw systems for maxillofacial reconstruction and internal fixation were studied including examination of chemical components, measurement of mechanical natures, biomechanical and biological evaluation, in combination with retrospective analysis of 319 cases fixed by means of plate-screw appliances. The results demonstrated that all experimental data have satisfied basically the design and clinical requirements, with the exception of a weaker curve-resistance and fixative stability of miniplate as well as lower surface wear-resistance of pure titanium. So, further improvement is necessary. Additionally, the function classification and indication of plate-screw systems are discussed. The duration and condition of intermaxillary fixation is suggested. The complications and their causes are analysed through comparison among different fixation systems. PMID- 10677953 TI - [Image analysis of cell nuclear morphology and DNA content in osteosarcoma of the jaws]. AB - The measurement of nuclear morphology and quantitative analysis of nuclear DNA content were made by Interactive Image Analysis System in 47 osteosarcomas of the jaws. The results showed that with a decreased degree of differentiation, the cell nuclei of osteoblastic and chondroblastic osteosarcomas came to be large and irregular, those of fibroblastic osteosarcomas, however, to be small and round. It was revealed that high-grade osteosarcomas had increased amounts of DNA and that the mean of DNA content, proportion of hyperploid cells and tumor ploidy were found to be of important prognostic value. PMID- 10677954 TI - [Studies on aging enzyme activities of the human dental pulp blood vessels]. AB - Histochemical distribution and age changes of alkaline phosphatase (AKPase E. C. 3. 1. 3. 1) and adenosine triphosphatase (ATPase E. C. 3. 6. 1. 3) in normal human dental pulp blood were investigated qualitatively and quantitatively. 42 dental pulp samples from patients aged 10 to 70 years old were divided into three groups. Results showed that accompanying aging, activities of AKPase and ATPase of dental pulp blood vessels decreased. AKPase staining quantities for group one (10-30 years), group two (31-50 years) and group three (51-70 years) were 40.62 +/- 13.79, 36.75 +/- 9.78 and 19.20 +/- 5.35 and ATPase staining quantities were 33.00 +/- 10.09, 27.53 +/- 16.60 and 23.27 +/- 5.04 respectively. The endothelial cells of capillaries in dental pulp changed greatly. With aging, metabolic ability of dental pulp decreased gradually. In addition, it was observed that the capillaries in sub-odontoblasts decreased and odontoblast layer became thin with aging. The close association of the capillaries of the pulp to the odontoblasts had been demonstrated. PMID- 10677955 TI - [Gama hydroxybutyrate sodium (gama-OH)-diazepam combined anaesthesia for cleft palate repair in children below 3 years old]. AB - To investigate the effect of Gama-OH-Diazepam combined anaesthesia for cleft palate repair in children below 3 years old. METHODS: Heart rates were measured in 120 children at induction, at 5 minutes after endotrocheal intubation, at finishing of operation, and at 5 minutes after extubation. In addition, changes of SpO2 was randomly detected in 10 children. RESULTS: Heart rate decreased after intubation, increased when operation was completed and decreased again after extubation. SpO2 of all stages were in the normal range. CONCLUSIONS: This method of anaesthesia is suitable for cleft palate repair in children below 3 years old, with the advantages of little effect on the children's circulatory and respiratory systems, and being easy to perform and control. PMID- 10677956 TI - [Palatography including lingual and occlusal surface of teeth, palatogram pattern of Chinese sounds]. AB - In order to develop a method that can determine not only the contact pattern of the tongue with the palate but also with the teeth during speech. Static palatograph recording system was used in this study. Palatograms of six normal dentulous adults were made by the recording system during pronounce the following Chinese phonetic alphabets: /S/, /X/, /D/, /J/, /N/, /K/, /R/, /L/. The contact pattern that is the contact area of tongue to palate and tongue to teeth in the palatograms were identified and analysed. The result indicated that the palatograms of the six normal adults are different which can be divided into four types: Type1/S, X/; Type2/D, N, J/; Type 3/K/; Type 4/R, L/. PMID- 10677957 TI - [The inhibitory effect of hydroprednisolone on tumor necrosis factor in rabbits' temporomandibular joint disturbance syndrome]. AB - In order to study the mechanism of hydroprednisolone for temporomandibular joint disturbance syndrome (TMJDS), 12 white rabbits were injected TNF into the rabbits' right TMJDS for 2 to 3 times, 90,000 u each time, then another drug, 2 mg hydroprednisolone, was injected into these TMJDS again. The rabbits were killed after 8 to 18 days and the TMJDS were examined by microscope. We found the articular tissues only destroyed slightly and sometimes the cartilages were nearly normal. So, the study suggested that hydroprednisolone can reduce the destruction of TNF on rabbit's TMJ obviously. This may be the cause why hydroprednisolone is useful to TMJDS. PMID- 10677958 TI - [The use of panoramic radiograph to observe abnormal movement of condyle]. AB - The abnormal movement of mandible is one of the three chief symptoms of TMJDS, it is described usually by the degree and type of mouth opening. We use close and open mouth position panoramic radiographs of 144 cases with TMJDS to observe the relationship between the head of condyle and the articular eminence. It can show the different degrees of movement of condyle and the degree of movement of both sides to see they are in balance or not. We classified the movement of the condyle in to six types, and measure the degree of mouth opening on the same film. The rela-tionship between the result of them and the climnical symptoms are then analysed. We concluded that the supermovement of the condyle, which cause the acute and chronic trauma to the structures of TMJ and related muscles and ligaments, is one of the chief etiologic factors of TMJDS. The method and advantages of using panoramic radiography to observe movement of condyle are discussed. PMID- 10677959 TI - [A study of parotid salivary proteins from caries-free and caries-active people by high performance liquid chromatography]. AB - In this study, we isolated and analysed parotid salivary proteins of different individuals with different caries-susceptibility. Protein fractions isolated were identified by biochemical methods. The results showed that six proteins were isolated from parotid saliva by HIC (hydrophobic interaction chromatography). Four proteins were found in all hydrophobic interaction chromatography. Three of them were identified as different polymorphic forms of proline-rich proteins (PRPS) and the other as alpha-amylase. Comparing chromatography of fifteen healthy caries-free adults with ten caries-active adults, we found that there was no difference in salivary protein composition. Three different phenotypes of PRPS had different relations with caries-susceptibility and relative ratios of the three PRPS were different between two groups. It suggested that abnormal proportions of salivary proteins may be another important determinant in caries susceptibility. PMID- 10677960 TI - [A comparative study of autologous facial nerve graft in situ with the vein tubulation: the observation of neurons changes]. AB - For further study of the effects of the repairment of facial nerve with vein tubulation, we used horseradish peroxidase (HRP) retrograde mark method to observe the changes of the labeled cells in facial neucleus after the tubulation of rat facial nerve. And, we compared the changes with nerve graft in situ and the normal group. The results showed: 1. The vein tubulatiton can repair the short defect of facial nerve as well as the nerve graft does. 2. Irregular outgrowth of axon sprouts could be found in both methods. 3. There was few chance for homolateral target organ to be reinnervated by contralateral motoneurons in both methods. 4. Using the vein tubulation method, a more natural relationship between the peripheral target organ and central nucleus may be easily achieved. PMID- 10677961 TI - [Ultrastructure of adherence of Streptococcus mutans MT8148 and its glucosyltransferase deficient mutants]. AB - The ultrastructure of adherence of Streptococcus mutans MT8148 and its glucosyltransferase deficient mutants were compared by Scanning Electronic Microscope. Four patterns of adherence of tested strains in the presence of sucrose were observed. These results suggest that the cell-associated GTase is responsible for firm aggregation of cells. However, cell free GTase is also important in aggregation between cells. PMID- 10677962 TI - [Three-dimensional finite element stress analysis of magnetical retaining and bar retaining in overdenture implants]. AB - The purpose of this study was to determine which type of denture retaining implants can produce better biomechanical effect on the bone tissue. Two mechanical models of magnetically retained complete overdenture supported by osseo-integrated implant and bar-retained complete over denture supported by osseo-integrated implant were made. The peak stress in mandibular peri-implant bone tissue of the models were studied under the same loading conditions vertically and horizontally by three-dimensional FEM stress analysis. The results were as follows: (1) Both types of dentures retention can produce significant stress effect on the peri-implant and other regions of the jaw bone and (2) Magnetical retention is more beneficial to maintenance both of peri-implant bone tissue and long-term success of denture supported by osseo-integrated implant than bar retention does. PMID- 10677963 TI - [Observations on before and after employ of screw of implant denture with SEM]. AB - The loss of screw is one of the clinic complication in MDIC implant denture cases. The purpose of this study is to find the cause of loss of screw. The authors has studied MDIC implant denture with SEM before and after employ of the screw and found that the causes of screw loss were crevice corrosion and friction pressure. Crevice corrosion may promote the loss of screw. Base the above condition, it is necessary to select Ti-alloy screw and associat with anticorrosive measure in MDIC implant denture construction. PMID- 10677964 TI - [The application of dental stents in implant therapy]. AB - The improperiety of the location of implants would cause many difficulties to prosthetic procedure. The function, appearance and long-term health of implant denture would then be effected. Dental stents were used to resolve the above problems for 30 cases of implant denture involved 74 implants in present study. A radiographic stent with given diameter sphere can be used to eliminate distortion and determine the bone quantity. A surgical stent can express the prosthetic plan to surgeons during operation directly in the mouths of patients. It is very helpful to the surgeons to take account of both the condition of jaw bone and the future denture making. Holes, tunnels and windows on surgical stents can be used as the guidances of drilling positions and directions. The surgical stents may also be a useful reference to surgeons to make bone graft, do some changes in surgical plan and even cancel the implant denture therapy when the situation of jaw bone significantly differ from what the expectancy. PMID- 10677965 TI - [A clinical study on treating the obstructive sleep apnea syndrome with orthognathic surgery]. AB - When severe mandibular hypoplasia accompanying obstructive sleep apnea syndrome (OSAS) were treated by the orthognathic surgery, the results were quite successful not only in patients' profile, occlusion and functions, but also in the subjective and objective evaluations related to OSAS. The paper summarizes the methods and surgical procedures of treating OSAS with orthognathic surgery through the diagnoses and treatment of a group cases suffering from severe mandibular hypoplasia accompanying OSAS. Severe mandibular hypoplasia can cause not only the deformity and the disorder of occlusion, but also the OSAS. The advancements of mandible and hyoid with orthognathic surgery can improve the patients'prfile, occlusion and expand the patency of upper airway, so it can cure OSAS. PMID- 10677966 TI - [Anterior segmental maxillary osteotomies (ASMO)]. AB - Anterior segmental maxillary osteotomies (ASMO) were performed with both the Wassmund and downfracture methods in 15 patients and folloned-up regularly for 12 months The postoperative maxillary incisor edge point "Is" movements and complications were examined. The results indicated that the point "Is" movement was 1.66 +/- 0.77 mm postoperatively. No major complications occurred, The stability of anterior segmental maxillary osteotomies was found to be acceptable. PMID- 10677967 TI - [Dentin non-collagenous proteins and their induction of dentin cells]. PMID- 10677968 TI - [To raise critical care in China to a higher standard]. PMID- 10677969 TI - [Effect of recombinant bactericidal/permeability-increasing protein on pulmonary cytokine mRNA expression in rats following hemorrhage and resuscitation]. AB - To determine the possible mechanisms underlying beneficial effect of recombinant bactericidal/permeability-increasing protein (rBPI) on acute lung injury response to blood loss, we used reverse transcription polymerase chain reaction to measure pulmonary tumor necrosis factor (TNF), interleukin 6 (IL-6) mRNA expression in a rat model of prolonged hemorrhagic shock (4.00 kPa, 180 min) followed by adequate resuscitation. The results showed that systemic plasma endotoxin concentrations elevated rapidly after a 180-min hemorrhagic insult (P < 0.05), and TNF, IL-6 mRNA expression in the lung were significantly increased at 2, 8 hours after resuscitation respectively. However, treatment with rBPI resulted in almost neutralization of plasma endotoxin values, remarkable reduction of TNF, IL-6 mRNA levels following hemorrhage/resuscitation. Also, it was found that rBPI administration markedly blunted the increase in pulmonary Evans blue dye extravasation, concomitant with a significant decrease in lung myeloperoxidase activity compared with the control group (P < 0.05-0.01). These data suggest that local proinflammatory cytokine mRNA expression associated with gut origin endotoxemia may be an important mechanism contributing to the development of hemorrhage-induced lung injury. Treatment with rBPI is effective in inhibiting marked TNF, IL-6 mRNA expression and ameliorating acute lung injury secondary to severe hemorrhagic shock. PMID- 10677970 TI - [The effect of anisodaminum and dexamethasone on microcirculation, TNF, LPO and pathology in MODS]. AB - In order to evaluate the effect of anisodaminum and dexamethasone to the microcurculation, TNF, LPO and pathology in MODS, we first established MODS model using cecem ligature and perfuration of rabbit then divided it into four groups: anisodaminum, dexamethasone, combined anisodaminum and dexamethasone and control. After used the corresponding drugs we observed the change of microcirculation, TNF, LPO and organ pathology. The result showed that microcirculation improved, the serum level of TNF and LPO decreased, and the pathomorphological changes lessensed. It is suggested that the combined use of anisodaminum and dexamethasone may be a new way for treating MODS. PMID- 10677971 TI - [The nosocomial infection in ICU: characteristics and prevention]. AB - In 84 consecutive patients who were treated in ICU, 35 had microbiological 5 data. 117 strains (bacteria: 77; fungus: 40) were isolated from 24 of the 35 patients for 97 times. The positive rate was 68.6%, or 28.6% of 84 patients. The gram-negative bacilli were still the main source of nosocomial infection in ICU, especially the pseudomonas aeruginosa, clostridium welchii, and E. coli, but gram positive cocci, especially MRSA infection were also increased. Mycotic infection increased very rapidly. 12 of the 24 patients were found a complicated mycotic infection. Six of the 12 (50%) died. Candida tropicalis and candida guilliermondi replaced candida albicans as the main pathogenic fungus. Sputum is still the most important specimen source of positive culture, followed by wound drainage, blood, urine, central venous catheter as well as feces. It is indicated that the respiratory system is still the main focus of nosocomial infection, and mycotic infection is most dangerous in ICU. Regular monitoring of microbial flora changes, detection correct use of antibiotics by drug sensitivity test, adequate surgical drainage, early and prevention of mycotic infection as well as improvement of ICU design will prevent or minimize the nosocomial infection in ICU. PMID- 10677972 TI - [Prospective study of central venous catheter-related sepsis in critically ill patients]. AB - To evaluate the incidence of central venous catheter-related sepsis (CRS) in critically ill patients, we performed a prospective study of the central venous catheters (CVCs) in ICU of the Peking Union Medical College Hospital from Jan. 1995 to March 1996. Of 151 CVCs, 13 (8.6%) had CRS, with an incidence of 16.7 episodes per 1000 catheter-days. Presence of infectious focus at catheterization, catheter insertion site, duration of catheterization, and the decrease of body temperature after catheter removal correlated with definite CRS, while difficulty of insertion, body temperature at catheter removal, as well as the decrease of body temperature after catheter removal correlated well with no CRS. The study showed that CRS is a serious problem in critically ill patients. Careful manipulation of CVCs is a major determinant in reducing the incidence of CRS. PMID- 10677973 TI - [Clinical study of systemic inflammatory response syndrome and multiple organ dysfunction syndrome in critically patients]. AB - We defined the epidemiology of systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) in critically ill patients, and evaluated the procession from SIRS to MODS and the therapeutic strategies. 230 patients were studied prospectively until discharge or death. On admission, the morbidity rate of SIRS was 71.3%. The mortality rate of the patients with SIRS was 18.9%. MODS was developed in 65 patients (28.3%), and 33 patients dided (50.8%). In SIRS patients with non-infectious SIRS, sepsis and septic shock, the morbidity rates of MODS were 22.8%, 61.1% and 85.7%, and the mortality rates were 11.4%, 30.6% and 50.0% respectively. The outcome of critically ill patients may be improved if SIRS is early diagnosed and the body inflammatory response is reglulated properly. PMID- 10677974 TI - [Changes of plasma cytokines in patients with severe trauma and their relationship with organ damage]. AB - We investigated kinetics of plasma TNF, IL-6 and IL-8 and their relationship with organ dysfunction and endotoxemia in 17 patients with severe trauma in order to further elucidate the role of cytokines in the development of organ damage and their production mechanism after trauma. Plasma cytokine levels significantly increased in trauma patients, and their plasma TNF was increased earlier. The cytokines were positively correlated with ISS, cardiac and hepatic enzyme activities, index of renal function, and plasma endotoxin levels. It is suggested that TNF, IL-6 and IL-8 may participate in the development of organ damage after trauma, and its release might be related to massive endotoxin translocation into body at the early stage of trauma. PMID- 10677975 TI - [Total knee replacement of severe flexion contracture deformities greater than 60 degree]. AB - The technique of total knee arthroplasty for the patients with severe flexion contractures of more than 60 degrees is not clear. Recently, We have performed 37 total knee arthroplasties in 23 patients with flexion contracture of more than 60 degrees (average 77.97 degrees). Among them, 14 knees (37.9%) with flexion contracture of more than 90 degrees, and 7 knees (18.0%) with 90 degrees flexion fusion deformities. Significant improvements occurred after averaged 4.3-year follow-up. Complications occurred in four patients: three had transient peroneal nerve palsy, and one had temporary circulatory disturbance of the lower extremity. They recovered after conservative therapy. We consider that severe flexion contracture of more than 60 degrees is not a contraindication of TKR. Staged bone resection and thoroughly soft-tissue release of the posterior capsule and collateral ligament balance were the critical procedure. If necessary, additional distal femoral condyle resection with posterior cruciate ligment sacrifice can be considered. PMID- 10677976 TI - [The structural changes of cortical bone beneath plate after rigid platefixation and removal]. AB - We investigated the structural changes of bone under the plate after plate fixation and removal. Twenty-four New Zealand white rabbits were plated on their intact left tibiae with stainless steel plates and 4 animals served as controls. The plates were removed 2 months after implantation in 20 plated animals, of which, 4 were sacrificed immediately after plate removal and the other were killed in successive groups of 4 each at 1, 2, 3 and 4 months after plate removal respectively. The remaining 4 plated animals were killed at 6 months after implantation. Bone samples were harvested and prepared for scanning microscopic observation. Internal fixation with a rigid plate may lead not only to osteopenia of the cortex under the plate, but also to disorganization of the cortex, in which the mineral column and collagen fibers were oriented in a randomized pattern. The regional osteoporosis could recover gradually after removal of the rigid plate. However, the restoration of normal bone structure occurred later than that of the bone mass. Delayed restoration of bone structure might be one of the potential causes of refracture of the plated bone. PMID- 10677977 TI - [Long-term results in 31 patients on total hemispherectomy modified for infantile hemiplegic epilepsy]. AB - To overcome long-term complications of hemispherectomy, We modified its operative method. The modified hemispherectomy was performed for 31 patients between 1985 and 1992. These patients were studied for 7 years after operation. The results showed total control of seizures in 28 children with chronic epilepsy (90%) or near-total control in the others. No deaths or delayed complications were noted but improvement in behavior and hemiplegia. CT and MRI showed marked shift of the remaining hemisphere. On brain-stem auditory evoked potentials, the latency of peak I was not variant (P > 0.05). The method makes the insulation of the subdural cavity from the ventricular system more reliable, and climates the pathological conditions. PMID- 10677978 TI - [Coronary artery bypass graft with internal mammary artery in 53 cases]. AB - Coronary bypass graft (CABG) with internal mammary artery was used (IMA) in 53 cases. They were treated medically but not effective. More than one time myocardial infarctions occurred 44 cases, and 16 of them had complicated ventricular aneurysm. All of them had moderate hypothermia cardiopulmonary bypass except one, who was only subjected to anastomosis of left IMA to left anterior descending branch without cardiopulmonary bypass. The mean grafts of this group were 4.28 (lift ventricular aneurysmectomy was performed simultaneously in 4 cases). Operative death was occurred in 4 cases. Of the 35 cases which had been followed for 6 months to 1 year, 30 were free of symptoms, 5 got better and had more physical exertion. It is suggested that CABG with IMA is satisfactory. PMID- 10677979 TI - [Surgical consideration in congenital partial atrioventricular septal defect in 59 patients]. AB - We report surgical anatomic characteristics, timing of surgery, corrective methods of partial atrioventricular defect and prevention of complications. In the past 15 years, we operated on 59 patients with partial atrioventricular septal defect, associated with conduction abnormality in 39 patients, common atrium in 2, mitral insufficiency (MI) in 54, tricuspid insiufficiency in 29, andunroofed coronary sinus in 6. One patient could not wean from cardiopulmanary bypass and died during operation. After surgery, 5 patients had moderate residual MI, and 1 complete heart block. During 1 to 16 years of followup, all but 1 patient with class III (NYHA) had satisfactory results. PMID- 10677980 TI - [Management of acute arterial occlusion of limbs: an analysis of 105 cases]. AB - 105 cases of acute arterial occlusion of limbs were admitted to our hospital from march 1982 to September 1996. We compared the treatment results of the patients with acute embolism and acute thrombosis. The limb-salvage rate of 86.8% in patients with embolic occlusion was higher than that of 57.9% in patients with thrombotic occlusion (P < 0.01), while the mortality rate of 8% patients associated with thrombosis was lower than that of 15.7% with embolism (P < 0.01). The results of limb salvage and mortality were related to different etiology of acute arterial occlusion, embolism and thrombosis. PMID- 10677982 TI - [Factors for postoperative persistent hypertension in patients with aldosterone producing adenoma]. AB - We determined the factors for postoperative persistent hypertension in the patients with aldosterone-producing adrenal adenoma (APA) in 53 patients with APA who were followed up for average 3.1 years. All had normal serum potassium concentration postoperatively. Blood pressure was normal in 37 patients (69.8%) but 18.7/12.7 kPa or more in 16 patients (30.2%) with persistent hypertension. Also compared were sex, age, history of hypertension, effect of reducing blood pressure to antisterone, preoperative blood pressure, time of persistent hypertension, serum potassium concentration, aldosterone concentration in 24 hour urine, amount of PRA, and the type of operation. The results showed that an APA patient aged 50 years or more appears to have a great chance of persistent hypertension than an APA patient under age of 40 years, and the odds ratio is 3:1. There was a significant difference between the mean age for persistent hypertension and for normal blood pressure, and varioas response of reducing blood pressure to antisterone (P < 0.05). It is suggested that for an older APA patient and the patient without of reducing blood pressure to antisterone, there are other factors for hypertension such as renal veinlet change or renal interstitial lesions except for hyperaldosteronism. We recommend renal biopsy (using kidney puncture) at the operating table for those patients in order to understand pathological change and guide treatment after operation. PMID- 10677981 TI - [Prenatal diagnosis and early treatment of congenital urological deformites]. AB - 11 children were treated in our hospital from 1987 to 1990 for the congenital urological defomities. They were diagnosed prenatally and were treated early after birth, and 5 of them were compared with 28 children who had been treated in the same hospital in the same stage for hydronephrosis due to the congenital pelvic-ureter stenosis, that was not prenatally diagnosed. Analysis of a series of renal functional test, special methods and clinical study, showed the possibility of prenatal diagnosis and the necessary of early treatment of congenital urological deformities. PMID- 10677983 TI - [Quantitative electroencephalogram monitoring the depth of anesthesia during skin incision]. AB - We investigated the EEG parameter changes of skin incision during different depth of sevoflurane/nitrous oxide analgesia. 65 ASA physical status I patients (aged 34 +/- 12 yr) scheduled for elective abdominal surgery were studied. The tracheal of each patient was intubated and the lungs were ventilated. Patients were randomly assigned to one of three groups. Anesthesia was maintained with 1% (group I n = 25), 1.5% (group II n = 20) or 2% (group III n = 20) end-tidal sevoflurane concentration in 66% nitrous oxide. Each of the concentration levels was maintained for at least 15 minutes before surgical incision. The EEG electrodes were placed on each patient in a front-oparietal montage (Fp1- A1, Fp2 A2) referred to Cz. EEG was recorded during 3-min period before incision. Hemodynamic variables were also monitored. Inadequate anesthetic depth was defined as patient movement in response to a 5-cm skin incision. The ventilation was controlled to maintain normocapnia (PETCO2 5 +/- 0.04 kPa). The data were analysed using ANOVA, liner correlation analysis and t-test. A significant difference between EEG parameters (SEF, BIS,) and skin incision responsive rate were found among the three concentration groups (P < 0.01). There were no difference of hemodynamics among the three groups. Patients who moved at incision also had significantly higher SEF, BIS leveles compared to non-movers (P < 0.01). Quantitative EEG determinants were correlated well with the end-tidal sevoflurane concentration and were a useful predictor of patient movement in response to skin incision during sevoflurane/nitrous oxide anesthesia. PMID- 10677984 TI - [Early diagnosis and treatment of mesenteric venous thrombosis]. AB - From 1967 to 1995, we treated 16 patients with mesenteric venous thrombosis (MVT). Major complaint was abdominal pain, and 13 patients had predisposing factors. One was diagnosed by color Doppler ultrasonography and cured by anticoagulant, 14 were misdiagnosed, 7 died postoperatively, and 8 cured. Early diagnosis lies on predisposing factors, discordance of symptoms and signs, color Doppler ultrasonography, CT, and angiography. Early operation for removing the involved intestine with its mesentery and early application of anticoagulant are essential to decrease the mortality and recurrence. PMID- 10677985 TI - [Nutritional rehabilitation for patients with extreme short bowel]. AB - The prolonged parenteral nutrition and intestinal transplantation is considered as useful therapeutics for patients with extreme short bowel. However, both of them are still limited. We report the results of nutritional rehabilitation for patients with extreme short bowel. Three patients whose residual intestines were 65 cm, 75 cm and 30 cm respectively were treated with growth hormone, glutamine and a fiber-containing diet for 2-3 weeks. The nutritional status of patients and the absorptive ability of residual intestine in these patients improved obviously after the treatment. This nutritional rehabilitation regime offers a new potential method for patients with short bowel syndrome. PMID- 10677986 TI - [Liver resection after transcatheter hepatic arterial chemoembolization for hepatocellular carcinoma and curative effect analysis]. AB - To study the therapeutic result of hepatic resection for those hepatocellular carcinoma (HCC) shrunked after transcatheter hepatic arterial chemoembolization (TACE) in the patients with unresectable HCC, authors reported 59 patients with HCC. Among the 59 patients, the maximum diameter of the tumor was 5.6 to 20.0 cm prior to the first TACE, mean 9.43 cm. The patients underwent 1 to 6 times of TACE, mean 2.9 times. The tumor diameters were reduced to 3.29 cm prior to operations. The duration between the last TACE treatment and sequential resection varied from 1 to 7 months, mean 2.53 months. Of the 59 patients, 35 patients' serum alpha-fetoprotein (AFP) levels were elevated. AFP levels returned to normal after TACE treatment in 13 patients. Of the patients, liver segmentectomy, combined liver segmentectomy or partial liver resection was performed in 56 patients, left trilobectomy in 2 and left hemihepatectomy in 1. Tumor necrosis ranged from 40% to 100% pathologically and complete tumor necrosis occurred in 9 patients. Of the 13 patients with AFP levels decreased to normal, 9 still had microscopic living tumor foci. The 1-, 3- and 5- year survival rates were 79.7%, 65% and 56%, respectively. These results indicated that TACE treatment can provide chance of tumor resection for those patients with unresectable HCC and good results can be obtained. PMID- 10677987 TI - [Clinical application of expanded radical operation with improved Whipple technique to late pancreas head cancer]. AB - Studies on the expanded eradicative and modified operation for treatment in patients of late pancreas head cancer, so as to enhance the incised rate, survival rate and life quality. Elective laparotomy was done in 20 patients. Expanded eradicative operation was performed in 18 patients (portal vein excision 19, left outer leaf of liver 8, transverse colon 3). We have reconstructed the portal vein 6, modified the Whipple operation, reconstructed the elementary tract using Roux-Y interposed jejunum with orientatied intussusception of artificial papilla. The incised rate of expanded eradication was 90% without postoperative serious complications such as biliary and pancreatic leakage, massive hemorrhage and operative death. The longest survival time was 5 years. The two-year survival rate was 50%. Statistical analysis (t test and x2) showed signithicant differences (P < 0.001).: (1) The exsion of original carcinomous focus may relieve the patient's symptoms and can prevent portal vein from carcinomous cell infiltration. (2) After enbloc incision with metastatic focus, sequential treatment could be given successfully. PMID- 10677988 TI - [Serial modifications of bacon's pull through resection for low rectal cancer]. AB - Four modifications of Bacon's pull-thru resection were undertaken to improve postoperative defecatory control and to avoid the 2nd stage resection of the pull thru colon stump. 54 cases of modified Bacon's rectal resection (9.64% of sphincter saving resections) were performed between 1954-1989. The 1st modification (1954) preserves the levator ani and the intact anorectal ring. The 2nd modification (1964) preserves the dentate margin and anal transitional zone (acute anal sensation), thus greatly improving defecatory control. The 3rd modification (1980) simplified intraanal resective procedure. The 4th modification (1991) was to encircle and ligate the distal end of colon over a sterilized corrugated intraluminal splinting tube (The other end of which is already connected to a long latex tube prior to operation). The latex tube was pulled out thru anus, until the colonic ligature reaches the Dentate margin, to substitute for colonic pull-thru and to divert feces during & after operation, 4 8 fine stitches approximate the colon wall 1 cm proximal to the ligature, to the cut edge of anal mucosa. The colon wall distal to the ligature sloughed in 7-10 days, the proximal colon has partially healed to the raw areas of anal canal without infection. The 2nd stage colon stump resection is thus obviated and hospitalization shortened. The postoperative anal function was good in 86.66%. Modified Bacon's operation is indicated for very low rectal cancer when the rectal remnant above levator ani after adequate resection is less than 1 cm which is difficult for intraabdominal anastomosis. It extends the scope of sphincter saving operation. It is a good substitute for Park's coloanal anastomosis. PMID- 10677989 TI - [Treatment of atrial fibrillation using maze procedure by radiofrequency ablation]. AB - From May 1994 to May 1996, 20 cases of atrial fibrillation were treated by means of Maze procedure by radiofrequenncy ablation, at the same time 19 cases of these patients were complicated with rheumatic heart valve disease and valve replacement operations were perfomned, in the other case atrial septal defect was repaired. Yoshio Kosakai's operation route was adopted in radiofrequency ablation procedure. After operation 16 patients of atrial fibrillation resumed sinus rhythm (80%), in 4 casess of atrial fibrillation sinus rhythm was unsuccessfully restored, two patients remained atrial fibrillation, one patient was of atrial flutter, the other was of nodal rhythm. Short time was needed in radiofrequency ablation Maze procedure, average time increase of aortic clamping was 20.5 minutes, and there was no danger of hemorrhage related to this kinds of Maze procedure. During 7-10 days after operation, there appeared superventricular arrhythmia which might be related to ill-distribution of radiofrequency ablation, and interference of atrial electric activity. PMID- 10677990 TI - [Complications of osteotomy of vertebral arch and body in kyphosis]. AB - Among total 67 postoperative complications resulted from osteotomy of vertebral arch and body in 439 cases of kyphosis, massive bleeding accounted for 2 cases (0.46%); spondylolisthesis, 6 (1.37%); paraplegia caused by compressing of the spinal cord, 5(1.14%); root pain, 6(1.37%); loss of correction, 26(29.55%); leaking and subcutaneous accumulation of CST, 5(1.14%); incision necrosis, 16(3.64%); lifting of luque rod with peneatration through the skin, 1(0.23%). Causes, treatment and prevention of these complications have been discussed in this study. We emphasize that the operation should be done after the stable of ankylosing spondylcitis. The degree of correction by osteotomy at one-level should be limited. PMID- 10677991 TI - [Reconstruction of anterior cruciate ligament using bone-patellar tendon prepatellar periosteum free graft with impacting bone technique]. AB - In order to make full and effective use of patellar tendon (middle one-third) during reconstruction of anterior cruciate ligament, we designed bone-patellar tendon-tendon of quadriceps femoris free graft with impacting bone technique and 38 cases were treated by this method. Follow-up for 2 years and 7 months showed that the clinical results were good. The exellent and good rate was 89.7%. Clinical results indicated that the fixation of this reconstructive method was firm and grafts in the joint cavity were patellar tendon tissue. The anterior medial rotatory instability of the knee joint could be corrected with the same graft that have enough length. The experience of acute rupture of anterior cruciate ligament and the rupture of anterior cruciate ligament combined with injury of meniscus were reported. PMID- 10677992 TI - [Forty six cases of application of free femoris forelateralis flap in trauma]. AB - In serious scar or chronic ulcer caused by burn or trauma, skin graft can not be survival and the local flap can not be found in these cases. Also skin grafts can not improve function of these places. Free flap transplantation should be used in these cases. Femoris forelateralis flap has some advantages. It is hidden from view and there is no effect after it is taken. It has constant blood vessels which are easy anastomosed. So the flap is more widely used. Forty-six cases were given femoris forelateralis flaps (hand burn scar deformity 15 cases, foot scar 10, forearm or leg scar or tissue defect 8, leg or foot chronic ulcer 8, face or neck tissue defect or serious scar 8). Forty-five cases (97.8%), were survival. Only one case had necrosis. The key of survival is choice of the operative indication, excellent anastomosis technique, good fixation postoperatively, care observation, and timely handling. In vasculitis or phlebitis ulcer, the vessel condition is bad. In old people, especial with heart or vessel diseases, condition is also bad. We shoule be careful in these situations. PMID- 10677993 TI - [Treatment of posterior urethral break caused by pelvic fracture and inflammatory urethral stricture via laying three silicone tubers into urethra]. AB - We investigated a new approach for the treatment of posterior urethral break caused by pelvic fracture and inflammatory urethral stricture. 7-10 days after the operation of urethral realignment on patients with urethral break caused by pelvic fracture or after forced dilatation of inflammatory urethral stricture, three silicone tubers were laid into urethra and kept there for three months. In 66 cases of urethral break, the cure rate (58/66) was 87.9%, the improving rate (2/66) 3.0% and the failure rate (6/66) 9.1%. In 15 cases of inflammatory urethral stricture, the recovery rate was 100%. In this group of 81 cases, the cure rate was 92.6% and failure rate was only 7.4%. This new therapeutic approach proved to be simple, safe and effective. It could also dramatically reduce the infection or the stricture of urethra. PMID- 10677994 TI - [Use of appendix to restore a ureter: report of 2 cases]. AB - In two cases the appendix was utilised as a substitute for part of the left ureter. Case one used the appendix as a conduit from the upper left ureter to the right one to treat the megaureter, and the other one used the appendix as a connection to join the two transected openings of the left ureter to treat the obstruction caused by the chronic ureteritis of the left ureter. Long-term observation after the operation showed satisfactory functions of kidneys. We suggested that the appendix is an autologus tissue whose active peristalsis, little mucous absorption along with its easy performance make this method highly valuable and practical. PMID- 10677995 TI - [High-methylprednisolone treatment in acute cervical spinal cord injury without fracture and dislocation]. AB - We compared methylprednisolone (MP), surgical decompression and both in acute cervical spinal cord injury without fracture and dislocation. The study involved 32 acute cervical spinal cord injury without fracture and dislocation. MP was given to 8 cases (high-MP group) within 8 hours after injury. 12 cases (surgical group) underwent surgical decompression (anterior or posterior approach) within 48 hours after injury. 12 cases (combined group) were treated with MP within 8 hour of their injury and surgical decompression within 48 hours after injury. Neurological function was assessed using scores according to international standards for neurological and functional classification of spinal cord injury. The results showed that combined group were more effective than high-MP or surgical group, in the complete or incomplete spinal cord injury, and in motor and sensory. The risk of such complication as gastrointestinal bleeding or delayed wound healing is not significant with using high-MP. PMID- 10677996 TI - [Video image analysis of AgNORs distribution in the testicular tumor]. AB - For the purpose of exploring the relationship between AgNORs and the biological behaviour of the testicular tumor, the value of enumeration and image analysis of AgNORs has been investigated in 10 normal testicular tissues and 49 cases of testicular tumors. The major findings were: AgNORs value for number, area, mean area, large partical number (LPN) and roundness in normal, benign and malignant tumors had significant difference (P < 0.01). The former four parameters turned larger when the tumor became more malignant, and they were significantly higher in the good prognosis group than in the bad one. LPN was higher in stage III than in stage I and II (P < 0.05), no difference was found between stage I and II. Other parameters had no significant difference in each stage. These results indicate that AgNORs is helpfull in differentiating the benign and malignant testicular tumors, in determining the degree of malignancy and offering an additional prognostic indicator in testicular tumors. Partical numbers, area and LPN were more helpfull. It had no important value in differenciating each stage. PMID- 10677997 TI - [The role and mechanism of fatty acids in gallstones]. AB - 18 Chinese cases and 37 Japanese cases of gallstones were analyzed according to the procedure of extracting fatty acids in the gallstone. High perforamance liquid chromatography (HPLC), showed that the contents of total fatty acid and free fatty acid in pigment stones were obviously higher than those in black stone or cholesterol stone. The ratio free saturated to total saturated fatty acid is the highest in intrahepatic pigment stone and less in extrahepatic pigment stone, which significantly differs from the latter two kinds of stones. This may indicate that phospholipid in the bile catablizes into free fatty acids involved in the formation of stone under the action of phospholipase in the course of pigment stone formation. The action of phosphoipase A1 is more important than phospholipase A2. PMID- 10677998 TI - [Experimental study on the treatment of smoke inhalation injury with lung lavage and exogenous pulmonary surfactant]. AB - To investigate the prevention and treatment effects of lung lavage and exogenous pulmonary surfactant (PS) on endogenous surfactant system dysfunction and acute respiratory failure caused by severe smoke inhalation, Wistar rats were randomly divided into five groups: Group I, normal control; Group II, smoke inhalation; Group III, smoke + lavage + PS + mechanical ventilation (MV); Group IV, smoke + lavage + MV; Group V, smoke + MV. The lungs were lavaged with 30 ml/kg 0.9% NaCl containing 100 mg/kg PS or same volume of saline via tracheal catheter at 5 min after smoke inhalation, then the animals were placed on a ventilator for 4 h, and observed until 24 h postinjury. The arterial blood gas levels, lung water volume, static lung compliance (Cst), total protein and albumin contents in bronchoalveolar lavage fluid (BALF), surface tension properties of BALF, and fatality rate at 24 h were measured. Smoke inhalation caused a similar acute hypoxia and severe carbon monoxide poisoning immediately in all injuried groups. The animals in group II showed acute respiratory failure, serious high permeability pulmonary edema, and surfactant system dysfunction. The surface tension properties of BALF, Cst, and the oxygenation were significantly improved with lung lavage and exogenous PS treatment. The lung water volume, total protein and albumin contents in BALF were decreased dramatically in this group. The fatality rate at 24 h declined markedly only in group III. It was suggested that lung lavage and exogenous surfactant treatment trestores effectively endogenous surfactant function inhibited by smoke inhalation, improves lung function, prevents high permeability pulmonary edema and respiratory failure, and decreases the mortality in early stage after smoke inhalation injury. PMID- 10677999 TI - [The mutual benefit role of pancreas and liver in combined transplantation]. AB - We study observed the mutual benefits of pancreas and liver in combined hepaticopancreatic transplantation in rats. The result indicated that pancreas, when transplanted with liver, could survive for a significantly longer time (13.4 +/- 1.01 days) than it transplanted alone (9.2 +/- 1.14 days) (P < 0.05). The interstitial rejection was mild and its rejection grade was significantly different from that of pancreas transplanted alone (P < 0.05, chi 2 test). The liver, when transplanted with pancreas, regenerated with strong competence and contact structure morphologically compared with liver transplanted alone. We concluded that pancreas could be immunologically protected against rejection and liver can be nutritionalized by pancreas in combined pancreas and liver transplantation. PMID- 10678000 TI - [The hepatic ischemia/reperfusion injury in cirrhotic rats]. AB - We assessed ischemia/reperfusion injury in carbon tetrachloride induced cirrhotic liver as compared to normal liver in the rats. Hepatic vein nitric oxide (NO) level was measured by method of luminol chemiluminensence, and portal vein endotoxin level by limulus lysate with chyomogenic substract. In cirrhotic liver, instead of diminishing the hepatic vein NO level increased significantly after ischemia and remained high till 5 hrs postreperfusion. The portal vein endotoxin level was also increased but to a higher level than that of normal liver. In cirrhotic liver, ischemia/reperfusion injury is aggrevated as evidenced by higher level of endotoxin, increased generation of NO. PMID- 10678001 TI - [Maze procedure for chronic atrial fibrillation: electrophysiologic basis and therapeutic effects]. PMID- 10678002 TI - [Pay more attention to the diagnosis and treatment of gallbladder cancers]. PMID- 10678003 TI - [Retrospective analysis of 830 extrahepatic biliary carcinoma]. AB - We retrospectively analysed clinical data of 830 patients with extrahepatic biliary carcinomas treated in our hospital from 1956 to 1995. Among them, 601 had gallbladder carcinomas, and 229 cholangiocarcinomas. The results showed that extrahepatic bile duct carcinoma accounted for 6.77% of bile duct diseases. Gallbladder carcinomas and cholangiocarcinomas accounted for 72.4% and 27.6% of extrahepatic biliary carcinomas, while the mean age was 56.55 and 56.44 years old, respectively. The sex ratio (male:female) was 1:2. 54 and 1. 46:1. The operation rate and resection rate were 65.1% and 29.4% in gallbladder carcinoma while 66.4% and 20.4% in cholangiocarcinoma. The incidence rate and resection rate of extrahepatic biliary carcinomas were elevated. PMID- 10678004 TI - [The long-term results of surgical treatment in 103 cases of hilar cholangiocarcinoma]. AB - We studied characteristics including clinicopathology, operation style, late survival rate of 103 patients with hilar bile duct carcinomas treated surgically in our hospital between 1986 and 1996. The factors affecting surgical treatment and late result of hilar bile duct carcinoma were analyzed. Of the 103 patients, subjected to 66 (radical resection, 36; palliative resection, 30) were section, and 37 (internal drainage, 11; external drainage, 26) were not. The total resectional rate was 64.1%. Operative mortality was 2.9% for palliltive or noresctive groups. In the radical resection group, the 1, 3 and 5-year survival rates were 96.7%, 23.3% and 13.2%, and the longest survival time was more than 8 years, whereas in the palliative resection group, the 3-year survival rate was only 3.8% and no one survived for over 5 years. In the unresected group, the survival time of the patients with internal drainage was longer than that of the patients with external drainage, but their viability was not improved significantly. Most of them died within 12 months. We proposed the new clinical types for hilar bile duct carcinoma. The pathological type in all patients survived for over 5 years was welldifferatiated papillary or tubular adenocarcinomas. The secondary operative rate in these patients was high. This showed that the biological characteristic is very important for the long-term results of bile duct carcinoma after operation. PMID- 10678005 TI - [Complications of laparoscopic cholecystectomy in China: analysis of 39,238 cases]. AB - All Chinese articles published from November 1995 through April 1994 about laparoscopic cholecystectomy were identified through CMCC. From 600 titles, 105 were read and left for analysis. Other 21 articles were from the 6th Biliary Surgical Congress and this hospital. Severe complications of laparoscopic cholecystectomy were identified in 409 patients (0.4%), bile duct injury accounted for 0.32%, postoperative cystic duct leak and bile leak for 0.11% and 0.20% respectively. Peritoneal abscess and bowel injury occurred in 0.07% and 0.06% of cases, and postoperative hemorrhages were identified in 0.1% of the cases. Fourteen (0.04%) postoperative deaths resulted from operative injury. The data demonstrate that laparoscopic cholecystectomy is an operation which associated with low morbidity and mortality. But bile duct injury is still a problem, and complications of laparoscopic cholecystetomy can be minimized by improving operative procedure. PMID- 10678006 TI - [Effects of gallbladder ruptured under laparoscopic cholecystectomy on pulmonary function]. AB - To assess effects of gallbladder ruptured and bile running out under laparoscopic cholecystectomy on pulmonary function, we preprospectively studied 20 patients without gallbladder rupture (group A), 10 with gallbladder rupture (group B), and 10 open cholecystectomy (group C). Significant differences were found in the group B and C to all pulmonary function values compared with preoperative values until the 5th to 7th postoperative day (P < 0.05). The group A showed only a significant difference on the first day after operation (P < 0.05). There are no significant differences between groups B and C. Comparing the group A with the group B and C respectively showed a significant differences from the 1st to 7th day (P < 0.05). This study confirms that the group A is superior to the groups B and C. Postoperative peritonitis is also an important cause of pulmonary dysfunction to reduce small airway function. Performing LC, we should try to avoid rupture of gall bladder or hemorrhage to fasten the recovery of patients' pulmonary function and reduce the chance of pulmonary complications. PMID- 10678007 TI - [The comparison of conventional open cholecystectomy, laparoscopic cholecystectomy and minor-incision cholecystectomy]. AB - To study the clinical value of minor incision for cholecystectomy, compared conventional open cholecystectomy (300 patients, group A), alparoscopic cholecytectomy (300 patients, group B), and minor-incision cholecystectomy (300 patients, group C) with regard to duration (day) of operation, amount of intraoperative bleeding, complications, time (day) of gastrointestinal function recovery, intravenous infusion, and expense and time (day) of hospitalization. The results showed that the operating time, amount of bleeding, duration of intravenous infusion, time of gastrointestinal function recovery and hospitalization in group A were significantly different from those in group B and C (P < 0.05), but the incidence in group A was lower than that in group B. The operating time, amount of intraoperative bleeding, and the time of gastrointestinal function recovery and intravenous infusion in group B were similar to those in group C, but there was a high rate of intraoperative and postoperative complication in group B (1.66%). There was no complication in group C (P < 0.05). Comparison of the effects among group A, B and C showed that it was superior in group C to those in group A and B. PMID- 10678008 TI - [Detection of bacterial DNA from cholesterol gallstones by NP-PCR and its clinical significance]. AB - To search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture. We used nested primers polymerase chain reaction (NP-PCR) technique to amplify bacterial gene fragments were amplified in vitro from DNA extracted from cholesterol gallstones. Comparative 16S ribosomal RNA sequence analysis was used for elucidation of bacterial identification. The gallbladder gallstones of 30 patients were analyzed. Bacterial DNA was found in the stones of 26 patients. There was no difference either in cholesterol and water content or in harboring bacterial DNA of gallstones. E. coli-related DNA fragments were found in the stones of 8 patients (26.67%). Propionibacteria type DNA was found in the stones of 7 patients (23.33%). Stones of 2 patients (6.67%) harbored bacterial gene fragments with similarity of Streptococcus pyogenes. A more heterogeneous sequence collection was found in 7 patients (23.33%) and could be assigned to the multiple bacterial infections. Another stones of 2 patients (6.67%) had bacterial DNA with lower molecularweight which might be related to some unidentified bacteria. The results suggested that most cholesterol gallstones harbor bacterial DNA. It is important to determine whether these microorganisms are innocent bystanders or active participants in cholesterol gallstone formation. PMID- 10678009 TI - [Maze procedure for chronic atrial fibrillation associated with mitral valve disease]. AB - From May 1995 to October 1996, 20 patients with mitral valve disease underwent the maze procedure for chronic atrial fibrillation and mitral valve replacement or mitral valvuloplasty. Epicardial mapping data demonstrated that the large macrorecurre flutter circuit was located in left atrium (14/20) and complex fibrillation in right atrium (18/20) of the majority of patients. No early death occurred. 20 patients were followed up for at least 3 months (range 3-20 months) and 14 patients for at least 1 year after operation, sinus rhythm and atrioventricular synchrony were restored (100%), atrial fibrillation was not induced by electrophysiologic study. The right and left atrial transport function was preserved (100%) by Doppler echocardiogram tracings. One patient died from acute necrotic hepatitis, 4 and half months after operation. Cox maze procedure was modified, atrial flutter and atrial fibrillation never occurred. In this study, the electrophysiologic mechanism of chronic atrial fibrillation associated with mitral valve disease, indications of operation and clinical results are discussed. PMID- 10678010 TI - [VATS in malignant pleural effusions]. AB - Between November 1992 and October 1996, 20 patients with malignant pleural effusions were submitted to VATS under general anesthesia. The overall positive histological diagnoses were obtained through VATS (20/20). The malignant pleural mesothelioma in 8 cases and metastatic cancers in 12 cases were histopathologically confirmed. Talc pleurodesis was performed for all the cases, and 18 patients gained lasting pleurodesis and the other 2 patients did not. The failure of pleurodesis was due to that the lung could not reexpand to come into contact with the chest wall. Postoperative complications included transient fever and slight asthma in 2 cases, and the symptoms were relieved obviously after some simple therapy to symptom. In conclusion, VATS has provided a high positive diagnosis of patients with malignant pleural effusions not diagnosed by conventional methods and good results in pleurodesis, but some operating skills must be emphasized. PMID- 10678011 TI - [Atypical manigioma]. AB - Most meningiomas have typical clinical and CT features. But some meningiomas which have an atypical clinical and CT findings can lead to a wrong diagnosis. The authors have reviewed 123 hospitalized patients with meningiomas which had been pathologically diagnosed. Nine of them had atypical meningiomas according to their histories, symptoms, signs and CT findings. All of these patients were mis diagnosed as having gliomas or metastatic tumors. The authors analyed these cases carefully and find the clues that may help to do an accurate preoperative diagnosis. PMID- 10678012 TI - [Scalp expansion in treatment of scalp defect with skull exposure]. AB - We repaired the scalp defect and skull exposure with scalp expansion. The technique consisted of two stages. First, expander was implaned an then N. S injected to its enough volume. The size of the expander was estimated by the formula that 1 cm2 needs 4 ml expansive volume. The incision was made at normal skin. Second flap was transferred. The expander was removed and the external skull which was necrotic severed. There were five patient with eight flaps at all, followed up for 6 months to 1 year. The hair grew. The results were satisfactory. This method not only treated the disease, but also avoided alopecia. PMID- 10678013 TI - [Clinical observations on syringomyelia treated by syrinx-peritoneal shunting with "T" tube]. AB - We 22 cases of syringomyelia were verified by MR and treated by syrinx-peritoneal shunting with "T" tube. After the operation, the patients were reexamined by MR and B ultrasound and confirmed that their shunting was functional. Symptoms of the patients disappeared. But four patients got slight numbness in one leg or at the saddle area. One patient suffered from a difficult movement in one hand. With the exception of 6 patients, 16 showed that both of the newly and original symptoms improved significantly. Follow up for 2 years with regular MR and B ultrasound reexaminations was made. The patients resumed their work and study. MR and B ultrasound reexaminations showed that all of syrinxes collapsed significantly as the same as that after the operation immediately. These mean that this kind of operation is useful and effective for most syringomeylias. PMID- 10678014 TI - [Investigation on matrix degrading enzymes of lumbar intervertebral discs]. AB - Changes in the macromolecular matrix of the intervertebral disc may predispose to biomechanical failure of the disc. Such changes would involve extracellular enzymes capable of altering the collagen and proteoglycan of the disc matrix. In this study, tritium-labeled type I collagen was used as a substrate to estimate the activity of collagenase in the discs of 41 cases of lumbar disc protrusion (LDP) patients by surgical intervention. The annulus fibrous (AF) and nucleus pulposus (NP) were measured separately. 34 normal discs harvested by autopsy acted as controls. For estimation of relative neutral proteinase content of 6 normal and 16 degenerated lumbar discs, polyacrylamide gelelectrophoresis (PAGE), heat-denatured collagen as a substrate, and photo-density scanning with peak area autocalculating system were adopted. The results presented that both AF and NP of the normal discs had a similar lower collagenolytic activity and a very limited activity of neutral proteinase, while the degenerated discs showed a higher activity, especially in the degenerated NP. The extruded type of LDP got a higher collagenolytic activity in NP than that of the prolapsed LDP. The fact showed that the matrix degrading enzymes play a very important role in the process of lumbar disc degeneration. The difference of disc degeneration is the biochemical basis of different clinical types of LDP. Matrix degrading enzyme system is a very complexed multienzymatic system. Other neutral proteinases may join this system besides the collagenase. PMID- 10678015 TI - [Influence of human bone morphogenetic protein (hBMP) on the articular cartilage pieces cultured in vitro]. AB - To understand the influence of hBMP on the articular cartilage, we investigated the expression of osteocalcin (BGP) and hBMP, and calcium deposit in cartilage matrix induced by hBMP. Under the inducing of hBMP, chondrocytes in cartilage pieces begun to express BGP and hBMP. The hBMP promoted calcium deposition in the cartilage matrix, hBMP made some chondrocytes further differentiat into osteoblast-like cells. We consider that these osteoblast-like cells might be other source of osteoblasts in endochondral osteogenesis. PMID- 10678016 TI - [The experimental studies of immune response of antigen-extracted bovine cancellous bone grafting]. AB - Xenogeneic bone grafting is an alternative to autogeneic bone grafting, but the intense immune rejection makes its clinical practice limited. We performed lymphocytes proliferation assay, enzyme-linked immunosorbent assay, and histological observation to evaluate the levels of cellular and humoral immunity, and the tissue reaction to the grafting of BALB/c mice receiving fresh bovine cancellous bone (FCB), antigen-extracted bovine massive cancellous bone (MCB), antigen-extracted bovine granular cancellous bone (GCB). Lymphocyte proliferation was increased in an early phase of the grafting and persisted for a long period in FCB group. Meanwhile higher levels of specific antibody were detected. In MCB and GCB groups, lymphocyte proliferation and specific antibody production were not ascended and no significant difference was observed between MCB and GCB groups in these immunological responses. There were no appreciable histologic signs of immune or foreign body reaction both in MCB and GCB groups. The results suggested that MCB, GCB were characterized less antigenicity and can be used as osteoconductive material or a carrier of bone growth factors. PMID- 10678017 TI - [The etiologic role of thermal injury on the induction of NO and NOS in plasma, burned wounds and visceral organs]. AB - Using a 35% TBSA burned animal moedl, we investigated the levels of NO contents (nitritc concentration assay) and NOS (hemoglobulin absorbent optical density test) in the burned wound, circulation, as well as five visceral organs which inluded heart, lung, liver, kidney and intestine mucosa. Samples were obtained from scalded male Wistar rats at 1 hour postburn (PBh1)PBh3, PBh8, PBh12, PBh24, PBh48 and PBh72, respectively. Samples of sham burned rats were obtained as controls. The plasma level of NO concentration was decreased within 72 hours postburn especially at PBh3, 8, 12, 24, 48 (P < 0.05 to P < 0.01). This was in agreement with our findings in the ESR assay. The cutaneous NO cotents was 3.8 to 57.8 times higher than that of plasma and other visceral organs. It is thus possible for burned skin tissue to be an important sites in the production of NO. However, the cutaneous NO might have little systemic influences and can only be a local fator due to its short half-life. The close relationship of changes in the NO and NOS in five visceral organs postburn provided strong evidence on the function of NOS as NO premerase. PMID- 10678018 TI - [Drug resistance and its mechanism of intrinsic drug-resistant cell line GRC-1]. AB - In order to probe the characteristics of drug resistance and its mechanisms of renal cell carcinoma, drug-resistant spectrum of renal cell carcinoma cell line GRC-1 was detected by in vitro MTT colorimetric assay, the mechanism of drug resistance in GRC-1 was also studied by the methods of both immunocytochemistry assay and flow fluorescence cytometry. The results demonstrated that GRC-1 was cross-resistant to adriamycin, vincrinstine, etoposide and carboplatinium, both mdr1 gene product P-glycoprotein and GST-pi which was an isozyme of glutathione S transferases were expressed in GRC-1. The accumulation of net intracellular drugs of GRC-1 was less than that of drug sensitive breast cancer cell line MCF7, and the ability of pumping drugs out of cells was higher than that of MCF7. The results suggested that there is an intrinsic multidrug resistance in GRC-1 cell line, and both P-glycoprotein and glutathione systems play a role in the development of drug resistance for GRC-1. GRC-1 is an ideal target cell line for the study of drug resistance. PMID- 10678019 TI - [Possibility of gene therapy in orthopedics]. PMID- 10678020 TI - [The different influences of splenectomy plus ligation of pericardial vein and shunt on portal hypertensive gastropathy]. AB - A prospective and controlled study has been made to probe into the different influences of splenectomy plus ligation of pericardial vein (SPLPY) and shunt on portal hypertensive gastropathy (PHG). 26 patients with cirrhosis of liver and portal hypertension admitted from January 1994 to April 1996 were randomized into two groups. One group was treated with SPLPV (15 cases), and the other shunt (11 cases). Gastroscopy was made in all patients pre- and postoperatively to observe the changes of gastric mucosa. It has been shown that SPLPV exacerbate PHG (P < 0.05) and the shunt has an opposite effect (P < 0.05). We consider that it is necessary to modify SPLPV and reevaluate the clinical effects of the both. PMID- 10678021 TI - [Clinical study of expandable venous stents for treatment of Budd-Chiari syndrome]. AB - Expandable venous stents is a new method for treatment of Budd-Chiari syndrome (BCS). Ten cases of BCS underwent expandable venous stents in inferior cava vein or/and in hepatic vein, or/and intrahepatic portosystemic shunt according to the location and the extent of lesion. Expandable venous stents were successful in all patients. No serious procedure-related complications were observed. The symptoms and signs disappeared. Portal pressure and inferior cava vein pressure gradient reduced immediately after treatment. The blood in stents flowed quickly. During follow-up of 1-21 months, rebleeding was observed in one case. The stenoses of intrahepatic shunts and inferior vena vein respectively occurred in two cases requiring repeat intervention. Therefore, expandable venous stent is safe and effective in patients with BCS, and will be of great value in wide application and further study. PMID- 10678022 TI - [Clinico-transcranial Doppler sonography monitoring on vasospasm and delayed cerebral ischemia after resection of intracranial tumors]. AB - The occurrence of vasospasm and delayed cerebral ischemia after resection of intracranial tumor has not received extensive attention clinically, and is often misdiagnosed and improperly treated as surgical brain damage or brain swelling. Seventy-two patients with intracranial tumor were continuously monitored pre- and postoperatively by means of neurological assessment and transcranial Doppler sonography. Vasospasm was found in 35 (48.6%) patients (18 mild, 13 moderate and 4 severe vasospasm). No significant difference among age, sex, surgical approaches, pathological diagnosis, duration of surgery, amount of blood loss and transfusion during surgery were found, but significant difference was seen in cisternal hemorrhage on CT scan and the amount of blood in cerebrospinal fluid. The cause and features of postoperative vasospasm were discussed, transcranial Doppler sonography played an important role in the diagnosis of vasospasm. To decrease the amount of blood in basal cistern by microsurgery in preventing vasospasm and to differentiate vasospasm from brain swelling are helpful to confirm the coexistent or causal relation based on neurological assessment, CT imagine, transcranial Doppler sonography and ICP monitoring both in deciding therapeutic strategy and successfully controlling vasospasm. Nimotop played a key role in preventing brain damage from vasospasm and cerebral swelling. PMID- 10678023 TI - [Influence of idiotypic network regulation on renal transplant]. AB - Pretransplant sera from 68 patients were tested for antiidiotypic antibodies (Ab2) and anti-antiidiotypic antibodies (Ab3) by lyphocytotoxicity inhibitory and potentiating assay. In the first six nomths, 6% of the patients in the Ab2 group had had a rejection episode compared with 21% of the patients in the no antibody (P < 0.01), whereas patients in the Ab3 group had significant rejection episodes (67%) compared with no antibody group (P < 0.01). These results suggest that Ab2 can introduce recipients tolerating the graft in some degree and patients with Ab3 are of high risk of graft rejection. PMID- 10678024 TI - [The significance of use of anti-intrarenal artery spasm in renal allografts with HAR like manifestation]. AB - In the past, severe ischemia of unknown cause in renal allografts after restoration of renal blood flow was regarded as a sign of hyperacute rejection (HAR) or other irreversible lesions, so that the grafts were usually excised during the operation. From January, 1994 to April, 1996, 8 cases of renal allografts with ischemia as described above were encountered in our hospital. Measures of anti-intrarenal artery spasm (IRAS) were taken to those grafts. Except that three grafts of HAR were excised, ischemia disappeared an renal functions recovered in 5 grafts. It suggested that ischemia of the five grafts was caused by IRAS. Thus, IRAS should be taken into account for renal allografts with severe ischemia of unknown cause and early anti-IRAS is essential for the diagnosis and treatment of IRAS. PMID- 10678025 TI - [Diagnostic value of CD44 splice variants in urine exfoliated cells of bladder cancer]. AB - To study the early and non-invasive diagnostic value of CD44 splice variants in urine exfoliated cells of bladder cancer. We used reverse transcriptionpolymerase chain resection (RT-PCR) and southern blot hybridization to detect CD44 splice variants in exfoliated cells in 40 urine samples (20 bladder cancers, 20 non neoplastic controls), and compared with the results of urine cytology on the same set of samples. 90% (18/20) of the urine samples of bladder cancer showed overexpression of CD44 splice variants while none of the 20 controls did so. This method not only has a sensitivity of 90% (18/20) which was much higher than that of 65% (13/20) by using urine cytology, but also is non-invasive and comfortable. The results suggest that CD44 splice variants in exfoliated cells in urine samples are a new tumor marker for early and non-invasive diagnosis of bladder cancer. PMID- 10678026 TI - [Hepatic resection and hepatic arterial chemoembolization for primary liver carcinoma]. AB - Three hundreds of primary liver carcinoma (PLC) were subjected to hepatic resection and hepatic chemoembolization (HACE). Among them, hepatic resetion was performed in 106 cases, and HACE in 194. The two-stage operation after HACE was performed in 23 cases. Hepatitis B antigen was positive in 69.8%. PLC with cirrhosis accounted for 81.6%. The effect of hepatic resection was superior to HACE. The cause of early local recurrence after HACE can be a valuable therapy in the hepervascular PLC. However, HACE should be unsuitable for ischemic PLC. Quantitative estimation of hepatic resection of PLC with cirrhosis was dependent on 5 parameters (ALT, serum bilirubin, PT and R15TCG), and morphological changes of cirrhotic liver. PMID- 10678027 TI - [Primary curative incision in the treatment of perianorectal abscess]. AB - More than 50% of the patients with perianorectal abscess treated with traditional incisional drainage will lead to fistula formation postoperatively. We present a procedure of primary curative incision for treatment with perianorectal abscess without fistula formation. The result of primary curative incision in comparison with traditional incisional drainage in a randomized control study showed that the incidence of postoperative fistula formation and recurrent abscess was 2.56% in the former and 56.25% in the latter. The key points of the procedure were discussed in detail and the causes and prevention of the disease occurred after injection of sclerotic drugs for hemorrhoids were also discussed. PMID- 10678028 TI - [Experimental and clinical study on gastroesophagostomy after cardiectomy of gastric stump]. AB - 60 hybrid dogs, divided into 2 groups, were subjected to subtotal gastrectomy in B-I and B-II pattern respectively. 20 weeks later, resection of cardia and fundus ventriculi and the gastric remnant esophagus anastomosis was performed. There were more omentum blood vessel embedments about the gastrojejunostomic and gastroduodenostomic stoma and collateral circulation was plentiful. In 1980, there were 25 patients with cardiac cancer of the gastric stump with an average period of 13 years and 3 months following subtotal gastrectomy and a mean age of 59.1 years. 19 patients were subjected to exploratory thoractomy, 17 resection of cardia and fundus of gastric stump and gastroesophagostomy (9 to tunnel gastroesophagostomy and 8 end-to-side esophagogastrostomy). The largest tumor was 5 cm x 4 cm. Except for a death due to intestinal obstruction following operation, the others attained a good recovery. Resection of cardia and fundus of gastric stump and gastroesophagostomy can be carried out for patients with cardiac cancer for gastric stump, which is in line with the principle of conservation surgery. PMID- 10678029 TI - [Video-assisted thoracoscopic bullectomy for giant bullous emphysema]. AB - Giant bullous emphysema often has serious dyspnea, and is difficult to manage. Bullectomy through thoracotomy for this disease carries a substantial morbidity and mortality. The aim of this report is to investigate the feasibility and key techniques of video-assisted thoracoscopic bullectomy for giant bullous emphysema. From December 1995 to October 1996, 6 patients with giant bullous emphysema underwent bullectomy by means of video-assisted thoracoscopy. Giant bullae occupied at least 50% of hemithorax, and 4 of which occupied more than 90%. According to Hugh-Jones dyspnea criteria: grade II in 3 cases, grade III in 2 and grade IV in 1, four bullectomies were done by video-assisted theracoscopy alone. One bullectomy and one left pneumonectomy were performed by combination of theracoscopy and a 8 cm thoracic incision. All procedures were accomplished successfully. The operating time ranged from 65 to 150 minutes. There was no blood transfusion and perioperative complications. Lung function was significantly improved in all patients after surgery (all better than grade II). Thoracoscopic bullectomy for giant bullous emphysema is a technically feasible and safety procedure, especially for group I and group II patients. However, there are still many problems to be resolved in thoracoscopy for group III and IV giant bullous emphysema. PMID- 10678030 TI - [Diagnosis and treatment of mediastinal tumor by VATS]. AB - Twenty-eight patients with mediastinal tumors underwent thoracoscopic operation, among which 22 received the resection of mediastinal tumor including mediastinal cyst (9), teratoma (5), neurogenic tumor (5), thymoma (1), vagothyrophyma (1), and lipoma (1), and 6 patients received diagnostic biopsy of mediastinal tumor. The preliminary results were promising. The experience of thoracoscopic operation on mediastinal tumor are discussed in this article and VATS is regarded as a new diagnostic and treating method which is superior to conventional thoractomy on some kinds of mediastinal diseases. PMID- 10678031 TI - [Treatment of carinal resections for primary pulmonary carcinoma]. AB - Threeteen patients of lung cancer of center type underwent carinal resections in our department from January 1985 to December 1996. There were 3 patients, T3N2M0 and 10 patients, T4N2M0. Palliative resections were performed for 3 patients and radical resections, for 10 patients. Carinal resection with right pneumonectomy was done in 3 patients, part carinal resection with right pneumonectomy 6 patients, part carinal resection with left pneumonectomy 1 case, and part carinal resection with right sleeve upper and middle lobectomy 3 patients. There were no operative complications and deaths. The 3-year survival rate was 54% and the 5 year survival rates, 30%. One patient has survived for 8 years. The satisfied results of operative therapy were followed when no lymph node metastesis in mediastinum and mass could be resected. PMID- 10678032 TI - [Treatment of esophageal and cardial stricture with TiNi memory alloys srent]. AB - A thermal-shaped memory metal stent, which is made of Nitinal, used in 95 patients with malignant tumors and 35 patients with anastomosis stenosis tumor resection. All patients had no opportunities of surgical operation and no response to dilation radiotherapy, chemotherapy, lasertherapy and traditional Chinese medicine. Dysphagia ameliorated in all patients after stent therapy. The mean dysphagia grade varied from 3.71 +/- 0.45(range 3-4) to 1.08 +/- 0.59 (range 0-2)(P < 0.01). Complications occurred in 49 patients (31.5%), including chest pain in 36, local bleeding in 5, tumor overgrowth in 4, stent migration in 2, and food obstraction in 2. In summary, treatment with placement of a Nitinol stent is effective, safe and simple in our patients with dysphagia which were due to malignant esophageal and cardiac strictures and is feasible for some benign esophageal and cardiac strictures as well through our clinical practice and observation nearly three years. PMID- 10678033 TI - [Measures for decreasing the early mortality after valvular replacement cardiovascular surgery]. AB - To probe the effective measure for decreasing the early mortality after artificial valvular replacement. We analyzed the cause of death among 1215 patients receiving artificial valvular replacement who were admitted in our intensive care unit during 1990-1995. All 44 deaths were serious-illed patients with preoperative cardiac functions of 3-4 degree. The factors affecting surgical survival rate include: enhancement of patient's cardiac function before operation and prevention of cardiac arrhythmia; a clear operational view and myocardial protection, especially for reoperative patients; application of membrane oxygenator; treatment of LCOS without delay, including intraveously administered agents and IABP used promptly when necessary; prevention and treatment of postoperative cardiac arrhythmia; strict, intensive care and synthesized treatment. PMID- 10678034 TI - [Limb salvage and amputation in osteosarcoma: report of 31 cases]. AB - Three different types of operation were performed in 31 osteosarcoma patients, and some cases received chemotherapy. It revealed, wide resection of tumor supplemented with chemotherapy improved the survival and limb salvage rate enormously. Chemotherapy and wide resection were all absolutely essential. The life of prosthesis was long and most patients were satisfied with it. Chemotherapy and limb salvage can enhance the survival and life quality in patients with osteosarcoma. PMID- 10678036 TI - [Auxiliary partial orthotopic liver transplantation in rats]. AB - A rat model of auxiliary partial orthotopic liver transplantation (APOLT) was developed in this study. Under hemihepatic inflow and outflow occlusions, the recipient underwent 75% partial hepatectomy before the liver graft comprising 30% of the donor liver was implanted on the orthotopic site. Forty rat APOLT were successfully performed. The 5-day survival rate of the recipient was 87.5% and that of the graft was 75%. On the fifth postoperative day, the living graft weight increased by 100%, the liver cell showed an active proliferation with a diploid of deoxyribonuclease (DNA), and the liver cells in DNA-synthetic phase accounted for 22.6 +/- 2.75%, significantly exceeding 12.22% +/- 1.48% of the normal rat hepatocytes (P < 0.001). It may be concluded that the rat APOLT is a more ideal animal model. PMID- 10678035 TI - [Nutritional effects of glutamine-enriched parenteral nutrition on transplanted small intestine in the rat]. AB - We evaluated the preventive effects of glutamine-enriched pareneral nutrition on the atrophy and hypofunction of the transplanted small intestines in the rat. Wistar rats received jejunal isografts and central venous catheters for parenteral nutrition (PN) for ten days. Rats received either PN with 3% alanyl glutamine or the PN with isonitrogenous ballanced nonessential amino acids. Glutamine-enriched PN significantly increased mucosal villous height mucosal thickness, crypt depth and villous surface area compared with the non-glutamine PN. Normal enterocyte ultrastructure of the graft was maintained in the Glutamine enriched group. Atrophied microvilli and broken mitochondrial crista were observed in the control group. It was found that the amount of absorption of 15N glycine was greater at the 1st, 2nd, 3rd hour in glutamine-enriched group than that in the control. These results indicate that the glutamine-enriched parenteral nutrition can promote the proliferation of mucosa, maintain the ultrastructure and improve the amino acid absorption in the transplanted small bowel in the rat. PMID- 10678037 TI - [The effects of microwave heating on osteoinduction of demineralized bone matrix in rabbits]. AB - In order to study the change of the autograft demineralized bone matrix, twenty four new-Zealand rabbits were randomly divided into four groups of 2, 4, 8, 12 week by sacrified time, each group had 7, 7, 5, 5 rabbits. Autoradiography was done for 2 rabbits in 2, 4 weeks respectively. The tibiae were removed by amputation at knee joint level, then they were cleansed of marrow and periosteun, cut into six pieces in length of 0.8 cm heated with microwave. Demineralized bone matrix were made by defated and decalcified, and implanted into abdomenal walls autograft. The results showed that the implants heated at 45 degrees C 30 min had same osteoinduction as control group. The 65 degrees C 30 minutes heating may reduce implant's osteoinduction slightly. The osteoinduction of the groups heated 75 degrees C for 30-60 minutes were impaired severely. The implants boiled at 100 degrees C 30 minuts had no osteoinduction at all, 65 degrees C 30 minutes may be a "safty" limit when heat treatment on bone clinically. Otherwise, bone formation ability would be reduced and remodelling was delayed. PMID- 10678038 TI - [Advances in the etiology and pathogenesis of congenital megacolon]. PMID- 10678039 TI - [Advances in the chemotherapy of osteosarcoma]. PMID- 10678040 TI - [Emphasis on combined modality therapy of breast cancers]. PMID- 10678042 TI - [The correlation between tumor angiogenesis and lymph node metastasis in primary breast carcinoma]. AB - To study the relationship between tumor angiogenesis and lymph node metastasis in primary breast carcinoma. Agiogenesis was assessed by the microvessel density (MVD) and expression of vascular endothelial growth factor (VEGF) using immunohistochemical staining. Paraffin-embedded specimens from 70 patients with primary breast cancer who had undergone radical mastectomy from 1984 to 1985 were studied. Thirty-one patients had histologically proven positive axillary lymph node (N+). The axillary node negative (N-) group was composed of thirty-nine patients. Microvessels per 200x (as MVD) and VEGF positively stained cancer cells per 400x were counted with light microscope. MVD and VEGF expression were higher in tumors with N+ showed than those with N-, MVD and VEGF expression were higher in N+ than in N-. MVD and the expression of VEGF are highly correlated with metastasis in primary breast cancer, which may serve as a parameter for determining tumor biological, metastatic potential and prognosis. PMID- 10678041 TI - [Prognosis and adjuvant chemotherapy of axillary node-negative breast cancer patients]. AB - The significance of postoperative chemotherapy was studied in 439 cases of unilateral primary breast cancer which were proved histologically with no axillary lymph node metastasis. The survival rate was analyzed by life table method. The prognosis of node negative breast cancer patients was mainly associated with tumor size. 10 year survival rate of patients with tumor less than 3 cm treated operativel and by operation combined with chemotherapy was 92.60% and 94.13% respectively. If tumor size was larger than 3 cm, the ten year survival rate of the patients treated operatived was 79.89%, and that of the patients with combined theropy 96.02%. The prognosis was statistically different between the two groups. As to age, status of menopause, pathologral type, kind of surgery and status of ER, the prognosis of patients treated by combined therapy was better than that of patients treated by operation alone. The postoperative chemotherapy was not significantly beneficial to breast cancer patients with tumor less than 3 cm. PMID- 10678043 TI - [The prognostic values of FCM determination of primary tumor and axillary lymph node of the patients with breast cancer]. AB - Using Flow cytometry, we determined the ploidy and SPF on the primary tumor and axillary lymph node of 58 patients with breast cancer. Follow up for 5 years evealed that 23 patients had recurrence. 21 of them died. We conclude that the patients with aneuploid and high SPF have a higher relapce rate than those with diploid and low SPF, especially in those with advanced stage tumor or metastatic lymph node. It seems that ploidy and SPF of primary tumors affect more strong the prognosis of the patient than those of axillary lymph node. The probable cause of the recurrence of diploid tumor patients suggest post operation treatment for the patients with aneuploid cancer. PMID- 10678044 TI - [Therapy for 29 sarcoma of the breast]. AB - We discuss the best therapeutic method for sarcoma of the breast. The data of all cases treated from March 1964 to May 1994 were analyzed retrospectively. Of all cases, 7 died, 9 recurred, and 15 survived more than 15 years and 5 over 20 years. The follow-up time was 4 months to 25 years. Most sarcomas were pathologically proved. The best therapeutic method was operation. The sarcomas of all cases must be extensively resected including surrounding tissues. The recurrent sarcoma should be resected again. The axillary lymphnodes should not be eliminated when lymphnodes is not discovered. PMID- 10678045 TI - [Male breast cancer: experience with 42 cases]. AB - Data were collected on 42 men with breast cancer treated at department of surgery, cancer hospital in Shanghai between 1960 to 1996. We studied several clinical features and the importance of established prognostic factors. Observation for 76 months showed the 5 year survival rate was 57.1%, and the total survival was 64.3%. Prognostic indicator analysis showed that only axillary lymph node status proved to have a prognostic impact. Tumor size, age did not show any prognostic influence. Because of less cases, we can not use Cox's regression model to do multivariate analysis. PMID- 10678046 TI - [Micrometastasis of colorectal carcinoma in bone marrow with immunohistochemical staining]. AB - From April 1994 to April 1996, immunohistochemical technique was used to detect micrometastasis of patients with colorectal carcinoma with monoclonal antibody to epithelial membrane antigen (anti EMA). Positive cells were found in their bone marrow in 23 of 57 cases, including 14 in 34 patients with colonic carcinoma (positive rate 41%) and 9 in 23 patients with rectal carcinoma (positive rate 39.1%). The positive rate was 40.4%. Statistical analysis showed that the modified Dukes stage of the cancer, the pathological type of carcinoma and the age of the patients were correlated with the positive rate, but the primary locations of the cancer were not. This method can be used to identify the extent of the cancer and direct clinical comprehensive treatment for colorectal carcinoma. PMID- 10678047 TI - [The effect of low temperature on von Willbrand factor expression of cultured human umbilical vein endothelial cells]. AB - We investigated the effect of low temperature on von Willbrand factor (vWF) of endothelial cells (EC) and the importance of low temperature in thrombogensis. We used the immunocytochemistry staining and image analysis system to study the expression of vWF of cultured human umbilical vein endothelial cells in low (33 degrees C) and normal (37 degrees C) temperature conditions. The expression of von Willbrand factor of EC was higher in low temperature than in normal temperature (P < 0.001). The low temperature plays an important role in thrombogensis. PMID- 10678048 TI - [Nucleolar organizer regions enumeration in testicular teratoma]. AB - To differenciate the benign and malignant testis teratoma. We investigated the value of AgNORs enumeration in 10 normal testis tissues, 6 benign teratomas and 7 malignant teratomas. The shape of the AgNORs in normal and benign teratoma was round, regular, with clear boundary and even size, setting in the center or the margin of the nucleus. In the malignant teratoma, the shape was irregular with unclear boundary and uneven size, setting in the near center or scattered in the nucleus. AgNORs enumeration: normal group: 1.56 +/- 0.17, benign teratoma: 2.40 +/- 0.26, malignant teratoma: 5.24 +/- 0.36. There was significant difference between the two groups (P < 0.01). The range of AgNORs enumeration in benign and malignant teratoma was separate. These results indicate that the mathod is helpful in differentiating benign and malignant testicular teratoma, and may provide an objective parameter. PMID- 10678049 TI - [Expression of the bcl-2 and bax oncoprotein in TCC and its clinical significances]. AB - To investigate the role of bcl-2 (an apoptosis suppressing oncogene) and bax (an apoptosis accelerating oncogene) in the development of TCC. We investigated bcl-2 and bax expression by means of immunohistochemical technique in 34 cases of TCC and in 9 cases of normal bladder tissue. Bcl-2 was positive in 44.44% of normal bladder tissue and in 82.35% of TCC. Bcl-2 staining intensity was significantly stronger in TCC than that in normal bladder tissue. Intensity and positivity of bcl-2 also increased with increasing grades of TCC. Bax was positive in 88.89% of normal bladder tissue and in 52.94% of TCC. Bax staining intensity was significantly weaker in TCC than that in the normal bladder tissue. These results suggested that increased expression of bcl-2 and decreased expression of bax in TCC play an important role in the development of TCC. PMID- 10678050 TI - [Fertility and related hormones before and after female successful renal transplantation]. AB - To evaluate the level of hormones before and after female renal transplantation, we measured pituitary gonadal hormones, estrovite, progestin, and prolactin in 25 renal transplant recipients (RTR) and 25 cases of chronic renal failure (CRF) using engyme immunoassay (EIA). The results indicated that serum RRL, FSH and LH level were reduced in RTR females compared with CRF, whereas E2 and P were normal. Serum PRL levels were elevated in CRF females whereas P levels were significantly lower compared with those of other groups. After clomiphene stimulation test, the plasma levels of LH FSH and E2 elevated, suggesting hypothalamic anovulation. Following successful renal transplantation, uremic hypothalamic disfunction was ameliorated and normal menstrual cycle, fertility was restored. During dialysis, treatment was given using suit therapy rather than trigger the ovulatory. Renal transplantation is the best treatment. PMID- 10678051 TI - [Surgical management of patients with infected vascular prostheses]. AB - We evaluated the therapeutic efficacy of surgical management of patients with infected vascular prostheses. Eight cases of infected vascular prosthetic grafts from 250 prosthetic bypasses were reviewed. The rate of graft infection was 3.2%. Clinical manifestations were localized wound infection with prosthetic graft exposure, anastomotic hemorrhage and gangrene in lower extremity. Treatment included graft removal and debridement; graft removal and primary amputation; graft removal and revascularization; debridement and local graft irrigation. Two cases died from anastomotic hemorrhage and the others recovered. The predisposing factors of vascular prosthetic infection are diabetes mellitus, secondary hemotoma and reoperation in the same position. Conservative treatment efforts without revascularization justifies a more aggressive approach to suspected graft infection. PMID- 10678052 TI - [Diagnosis and treatment of congenital choledochal cysts in adults]. AB - We report the experience with 97 adult patients with bile duct cysts in the past 30 years (1965-1995). The median age at time of initial therapy in our department was 37 years (range, 16 to 78 years). Clinical symptoms in most cases were non specific, resulting in delayed diagnosis. 74 patients (76%) had coexistent pancreaticohepatobiliary disease. Carcinoma of the biliary duct occurred in 16 patients (17%). Abnormal pancreatobiliary duct junction (APBDJ) was found in 28 patients. 94 patients underwent a total of 169 biliarytract procedures. Cyst excision with Roux-en-y hepaticojejunostomy and internal drainage were the main procedures. The result of long-term follow-up shows that internal drainage frequently resulted in recurrent cholangitis and cyst malignancy which need reoperation. In contrast, cyst excision was associated with a significantly lower incidence of recurred cholangitis and lower occurrence of malignancy. Thus, cyst excision with Roux-en-y hepaticojejunostomy is recommended as the treatment of choice for adult patient. PMID- 10678053 TI - [Clinical application of transmyocardial laser revascularization]. AB - Transmyocardial laser revascularization (TMLR), a new technique, provides direct perfusion of ischemic myocardium via laser-created transmural channels. From 1995 to 1996, we treated 7 patients (mean age 60 years, all men) with TMLR. Preperatively, 5 patients were in angina class (C. C. S) 3-4 and 2 patients had unstable angina. To identify the location and extent of their reversible ischemia, the coronary angiogram, 99mSPECT and/or dobutamine echocardiography was performed before operation in 7 patients. Through a left anterior thoractomy in the fifth intehcostal space, heart exposure was gained. With the use of a 700 watt CO2 laser, TMLR was performed on the beating heart. An average of 22 +/- 3 channels were created in 45 minutes with a total operative time of less than 2 and half hours. The in hospital mortality was one of 7 patients. Follow-up ranged from 2 to 12 months (accumulated 48 patient-months). Postoperatively, the relief of angina was noted in 6 patients. Postoperative SPECT and dubatamin-UCG were obtained at 3, 6, and 12 months. (99mTc) SPECT showed a significant improvement of myocardial perfusion in the area of reversible ischemia. Dobutamin-UCG documented an increase in the ventricular wall motion and LVEF in 2 patients as compared with basline. These early results indicate that TMLR may provide angina relief, improve myocardial perfusion and increase cardiac function for patients with ischemic heart disease. PMID- 10678054 TI - [Experience and lessons of lung transplantation]. AB - In the last 2 years, four hospitals performmed cooperatively single-lung transplantation for 3 patients, 1 left lung and 2 right lung. The patients had pulmonary fibrosis, COPD with cancer or tuberculosis, infection in some extent before transplantation. The patients died from severe infection 9, 48 and 43 days after the operation. We discussed the selection of donors and recipients, operative procedures, postoperative management, especially monitoring, differential diagnosis and treatment of infection and rejection. HLA compatibility and transbronchial lung biopsy were important to the success of lung transplantation. We believe that rejection and infection are important causes of short-term death that should be given more attention. PMID- 10678055 TI - [24 hour esophageal pH monitoring in patients with cardiac cancer postoperatively]. AB - Twenty-four-hour esophageal pH monitoring was performed in thirty patients with cardiac cancer on the 13rd to 18th postoperative day in order to understand the condition of gastroesophageal reflux (GER), the value of commonly used operative procedures and the effect of patients sleeping position for preventing GER. GER parameters were higher than those of normal subjects (P < 0.001), but there were only 60% patients who had typical GER symptoms. No significant differences were found between anastomoses in abdomen and chest, encasing-in style, and "scarf" style (P > 0.05). Patients sleeping with upper body raised for 30 degrees had much lower GER parameters than those of controls. Manual anastomosis often used now couldn't decrease GER. All patients with cardiac cancer have postoperative GER though they had no typical symptoms of GER, but GER was avoided during sleeping with upper body erected. PMID- 10678056 TI - [Traumatic ascending ischemic injury of the spinal cord]. AB - We report five cases that the paraplegic level showed ascending from the segment of spinal injury upward to the higher level. In T10-11 fracture-dislocation, the paraplegic level ascended gradually to C2-3 in case 1 within 10 days and death due to breathing paralysis. In case 2, the paraplegic level ascended to cervical cord with loss of strength in both arms within 14 days. The remaining 3 cases were Ti2(in 2 cases) and L3(1 case) fracture. Their paraplegic level ascended to T8(2 cases) and T9(1 case). There were lower motor neuron paralysis in all cases. Observation on spinal cord specimen of case 1 showed thrombosis of dorsal vessels at T9-10 segment and multiple thrombosis were found in anterior spinal vessels, central spinal vessels and intramedullar small vessels upward to C3 and downward to S1 segments. The whole cord underwent ischemic necrosis. PMID- 10678057 TI - [The vascular basis of pedicled adrenal transposition and its usage in the treatment of Cushing's disease]. AB - To treat Cushing's disease, we studied the length and exterior diameter of the inferior phrenic and superior adrenal artery and vein, and the reflow of the adrenal vein. The length and exterior diameter of those blood vessels were divided and measured by veiner calipor. Six patients with Cushing's disease were treated by pedicled adrenal transposition to the musculus of the back. The right inferior phrenic artery was 75-110 mm from the exterior edge of the vena cava to the distant branch of the superior adrenal artery, while the left inferior phrenic artery was 70-90 mm from the left edge of the aorta to the distant branch of the superior adrenal artery, both of which have veins to accompany them. The superior adrenal artery came from the inferior phrenic artery relatively common and was divided into six branches. The exterior diameter of the distal superior adrenal artery was 1.5-2.5 mm and could be freed for 40-80 mm. Lateral veins could complete the adrenal blood drainage after the left central adrenal vein was ligated. The transpositioned blood vessel pedicle consisted of the distant inferior phrenic artery and vein, the distant branches of the superior adrenal artery and vein. The patient who had received the transposition treatment had neither hypocortisolism nor recurrence after 4 to 35 months. The length of the inferior phrenic, superior adrenal gland vessels could content the remained adrenal gland to reach their transposition bed when freed. The pedicled adrenal transposition to the musculi is an ideal method in the treatment of Cushing's disease. PMID- 10678058 TI - [Experimental study on the effect of extremities ischemia reperfusion inducing anterior tibial compartment pressure]. AB - We observed the changes of compartment pressure after extremities were reperfused at different intervals of ischemia and its relationship with the changes of serum MDA and CPK content. The protective effect of mannite on extremities ischemia reperfusion was also studied. 18 New Zealand rabbits were separated into three groups: group A (ischemia 4 h); group B (ischemia 8 h); group C (ischemia 4 h). The rabbits of group A and B were intravenous injected with normal saline (NS), and group C was injected with 20% mannite. The changes of compartment pressure of serum MDA and CPK content and the histological changes of skeletal muscles in every group were noted at different times: preoperation; the beginning of ischemia; the end of ischemia and reperfusion for 4 h; 24 h; 72 h, respectively. The longer ischmia lasted, the more serious the tissue damage was. Reperfusion made tissue damaged further. After reperfusion for 24 h, the damage was the most serious. Meanwhile, the serum MDA and CPK content also reached the peak value. These changes were in accord with those of compartment pressure. On the other hand, the damage of mannite group was relatively mild. Extremity ischemia reperfusion would led to the increase of compartment pressure, then intensified the damage. The damage was the most serious at 24 h of reperfusion. Using mannite could reduce the tissue damage and compartment pressure. Mannite should be recommended as the first-selected drug for extremity ischemia reperfusion injury. PMID- 10678059 TI - [Lumbar intervertebral disc injuries]. PMID- 10678060 TI - [Prevention and treatment of joint prosthesis complications]. PMID- 10678061 TI - [Severe vascular injuries following total hip replacement]. AB - Severe vascular injury following total hip replacement (THR) is very rare. It has an a incidence of 0.25% and poses a serious threat to the involved limb and the patient's life. Vascular structures affected include the external iliac artery and vein, femoral artery and vein. In 721 cases of THR 3 had vascular injuries. The first case sustained the injury of the external iliac vessel which was cut by the sharp edge of bone cement while removing the displaced prosthesis. The second had laceration of the iliac vein while a pneumatic drill was used in the anterion superior acetabulum, and the third had the cut injury of the femoral artery during resection of the capsular scar tissue. The understanding of the mechanisms and location of vascular injury during THR will help surgeons in early diagnosis and treatment of the injury. PMID- 10678062 TI - [One-stage reimplantation for the salvage of total knee arthroplasty complicated by infection]. AB - One-stage reimplantation for the salvage of infected total knee arthroplasty in 8 patients was reviewed at an average follow-up of 20.1 months late infections occurred in 7 (87.5%) patients. The timing of the diagnosis of the infection after knee arthroplasty was of the prosthesis averaged 11.5 months. No one had recurrent infection and pain was relieved significantly in all patients. Our results suggest that one-stage reimplantation is a reasonably reliable procedure for the management of an infected prosthesis. The use of Gentamicin-impregnated bone cement and the Streptomicin bead mixed with Gentamicin improve the success of treating or preventing recurrence of the infection. Early one-stage reimplantation was needed as soon as the deep infection was defined in order to decrease more destruction of the bone. PMID- 10678063 TI - [Polyethylene wear in periprosthetic tissue of failed cemented total hip arthroplasties]. AB - To elucidate the relationship between polyethylene wear and loosening of total hip arthroplasty, we observed grossly the periprosthetic tissues, polyethylene wear debris and polyethylene acetabular liners retrieved from 6 failed cemented total hip arthroplasties during revisions by means of polar light miscroscope and scanning electron microscope. The production mechanism of polyethylene wear debris from acetabular liner was analyzed together with its role in periprosthetic fibrosis and prosthetic loosening. Results showed the abrasion of two bearing surfaces interposed with third-body particles as called three-body abrasion was the main model of polyethylene wear. All inner surfaces of the liners appeared clear abrasive scratches and big flat debris were found in most of the retrieved tissues. Histologically, the wear debris of various sizes embraced by 1 or several polynuclear giant cells that were found not only great in number but also active in function. The implication is that three body abrasion is the key factor of high rate wear of the polyethylene liner and that polynuclear giant cell is acting as an important cellular component leading to the prosthetic loosening. PMID- 10678064 TI - [Total knee replacement in diabetic patients]. AB - From December 1987 to March 1995, 16 TKRs were performed for 9 diabetic patients at our department. 7 of them had rheumatoid arthritis (RA), and 2 osteoarthritis (OA). There were 1 male and 8 females. The average age was 55.9 years (range 49 69 years), and the average weight was 64.5 kg (range 54-78 kg). According to the hospital for special surgery (HSS) knee rating scale, the pre and postoperative evaluations were made. The HSS scores were improved after the operation from average 30.2 points preoperatively to 78.2 points postoperatively. The excellent and good rate was 94%. After 3.9 years follow-up (range 10 months to 8 years), the HSS score was 74.4 points, and the rate of excellent and good was 87.5%. Compared to the other 209 RA or OA patients with 287 TKR at the same period, the HSS score was almost the same, but the infection rate was higher (3.5%). We conclude that the TKR of diabetic patients could also get the similar results as common patients if the patients are under the good control of glycemia and medical treatment. PMID- 10678065 TI - [Sonographic measures of the hip joint of 1328 newborns of Han and Uygur nationalities-relative factors of congenital dislocation of hip]. AB - The four sonographic methods (Engesaeter, Terjesen, Graf, Morin-Harcke) were used for examining the hip joint of 1328 newborns of Han and Uygur nationalities. The difference between Han and Uygur newborns was not statistical significant, but some literatures reported that in our country, the incidence of different nationalities in different regions varied. Our conclusion is that there is no obvious relationship between the depth of the acetabulum and the percentage of the femoral head covered by the acetabulum. It is caused by other factors, inherent or acquired. The value of the hip joint of the male and female has significant difference (P < 0.0005). The incidence of CDH in girls is much higher than that in boys. This may be related to the different depth of the acetabulum and different percentage of the femoral head covered by the acetabulum. In the period of newborn, girl's depth of the acetabulum is smaller than that of boys. PMID- 10678066 TI - [Effects of peripheral biofilm infection of artificial prosthesis on bacteria]. PMID- 10678067 TI - [Posterior cruciate ligament injuries]. PMID- 10678068 TI - [Prognosis prediction of S-phase fraction and p53, c-erbB-2, estrogen receptor, progesterone receptor in axillary node-negative breast cancer]. AB - The prognostic factors in 180 axillary node-negative breast cancer patients with more than 5 year follow-up were searched for with multiple regression. Based on the regression results, the cases with treatment failure in 5 year and the cases with 5 year disease free survival were matched at 1:2 ratio. Then the c-erbB-2 protein, p53 Protein, estrogen receptor (ER), progesterone receptor (PR) and the S-phase fraction (SPF) were measured in paraffin-embedded breast cancer tissue. The results were analysed with log-rank test and Cox-Model. Among the biobehaviour factors measured, the SPF was the strongest prognosis predictor for ANN. The 5 year disease free survival rate of ANN with SPF < 10% or > 10% was 94% and 52% respectively (P < 0.001). The treatment failure relative risk of patients with SPF > 10% ANN was 11.31. The relative risk of other four factors was respectively, PR 3.58 (P < 0.002), ER 2.93 (P < 0.05), p53 1.44 (P > 0.3), c-erbB 2 1.38 (P > 0.5). The combination of PR or EP with SPF could make the relative risk even higher. PMID- 10678069 TI - [Application of preoperative angiography in diagnosis of gastric cancer]. AB - To improve preoperative diagnosis of gastric cancer, we applied DSA in clinical diagnosis. 36 patients with gastric cancer were observed. Preoperative DSA and postoperative pathological analysis revealed the following results. With DSA, the accuracy of diagnosis of invasion depth of gastric cancer reached 72.2%. We summarised the criteria for diagnosis of gastric cancer. If subsidiary anastomotic channels narrow or obliterate in DSA, the case may be early cancer. If parietal branches narrow or obliterate in DSA, the cancer may invade the muscle of the stomach. If arcade of this lesser curvature narrows in DSA, the cancer may invade the serosa of the stomach. If the roots of main vascular vessels, such as the left gastric artery, and vascular vessels of other organs narrow or obliterate in DSA, the cancer may directly invade other organs as well. Two-thirds of gastric cancer could be identified through DSA with their shapes and characteristics. The varied shapes of vascular vessels caused by invasion of gastric cancer could reveal growing mode of the cancer. PMID- 10678070 TI - [Extra-anatomical bypass for subclavian artery occlusion]. AB - 20 patients with subclavian artery occlusion were treated by extra-anatomical bypass from 1989 to 1996. There were 9 men and 11 women, aged from 20 to 63 years with a duration of 1 month to 18 years. The main causes of illness were Takayasu's arteritis and atherosclerosis. Symptoms of upper extremity ischemia were present in 17 patients and vertebrobasilar insufficiency in 11. Carotid subclavian/axillary bypass and axilloaxillary bypass were performed on 17 and 3 patients respectively. There were no operative deaths. Relief of symptomes was achieved in all the patients except one who had reocclusion of bypass graft due to rather poor outflow vessel. We stressed the surgiacal technique of extra anatomical bypass in the treatment of subclavian artery occlusion. With few postoperative complications and high patency rate, we consider extra-anatomical bypass a safe, simple, well-tolerated and durable procedure for the treatment of subclavian artery occlusive disease in high-risk patients. PMID- 10678071 TI - [Treatment of malignant or aggressive bone tumors with microwave induced hyperthermia]. AB - Limb-sparing procedures have been well established for dealing with malignant bone tumors. Unfortunately, these procedures have different problems. We used an alternative operation combined with microwave-induced hyperthemia to modify the surgical methods. Thermotherapy with microwave intracorporeal irradiation was used to treat 112 patients with bone tumors. In this series, 79 had malignant tumors and 33 aggressive benigh tumors. Postoperatively, immune therapy was carried out regularly. The patients immunologic functions were monitored by assay of the subpopulation of T cells, IL-2 and sIL-2R (soluble IL-2 receptor). Follow up varied from 3 to 50 months (mean 23 month) s. Excluding 5 patients with malignancy in the vertebrae treated for palliation, 107 were evaluated by oncological and orthopedic criteria. 10 patients had local recurrence and required amputation. The remaining 97 had excellent local control. In 12 of the 74 patients with malignancy of the extremities, lung metastasis occurred 4 months to 2 years after surgery. Pathological fracture occurred at devitalized bone in 8 patients. In 29 out of 40 tumor-free cases followed for more than 2 years, the knee joints functioned properly with almost full range of motion. Single photon emission computered tomography (SPECT) study revealed revascularization of the devitalized tumor bearing bone segment could accomplish in one year or more. The immune state was improved after thermotherapy plus immunotherapy in the majority of patients. These results indicated that the use of microwave hyperthermia and adjuvant immunotherapy in the surgical treatment of bone tumors can be considered a definitive procedure, which is safe and well-tolerated. PMID- 10678072 TI - [Tracheobronchoplasty: a report of 40 cases]. AB - Tracheo bronchoplastic surgery with or without simultaneous arterioplasty was porformed in 40 cases of various bronchopulmanary diseases (benign in 8 and malignant 32). The operation consisted of reconstruction of trachea in 3 cases, plastic carinoplasty 2 cases, and bronchoplasty and pulmonary arterioplasty 1 case, sleeve resection of bronchus 34 cases, 4 cases (10%) had postoperative complications. Two cases (5%) died shortly after operation. Thirty-eight cases recovered. Thirty cases of malignant tumor were followed up from 1 to 10 years. The survival rates were 83.3% (25/30), 53.3% (16/30), 40% (12/30), 23.3% (7/30) at 1, 3, 5 and 10 years respectively. We discussed indication, anesthetic management, surgical procedure and results. The indication must be controlled strictly. The operative procedure and postoperative management must be further improved because the complications and mortality of tracheobronchoplasty are higher than those of conventional resections. PMID- 10678073 TI - [Surgical treatment of non-thoracotraumatic pneumomediastinum]. AB - The experience of surgical treatment of non-thoracotraumatic pneumomediastinum in 48 cases was summarized. The etiology, classification, pathology, and surgical treatment of non-thoracotraumatic pneumomediastinum were discussed. We conclude that most non-thoracotraumatic pneumomediastinum was complicated by secondary diseases. Therefore, the effective treatments for primary diseases are important. A satisfactory effect must be based on different surgical techniques according to the different clinical symptoms and age of patients. PMID- 10678074 TI - [Diagnosis and surgical management of 22 patients with congenital coronary artery fistula]. AB - Twenty-two patients underwent surgical treatment of coronary artery fistulas. The right coronary artery was the most common vessel of origin (68.2%), and the most frequent drainage site was the right ventricle (45.4%). The operation was performed with the use of extracorporeal circulation in 21 patients. Only one patient underwent distal ligation without the use of extracorporeal circulation. The absence of operative mortality and severe postoperative complication provide clear indications for surgical treatment. In view of currently improved and standardized techniques of extracorporeal circulation, we believe that it should be employed routinely. PMID- 10678075 TI - [Simultaneous coronary artery bypass grafting with other cardiovascular surgical procedures]. AB - To improve the effect and reduce the mortality of the simultaneous coronary artery bypass grafting (CABG) with other cardiovascular surgical procedures, from Nov, 1984 to July, 1996, 51 patients underwent such operation. Among them 45 patients had valvular heart diseases, 4 postinfarction ventricular septal defect and ventricular aneurysm, and 1 myxoma of left atrium and abdominal aortal aneurysm. The operative mortality was 5.85% (3/51), and 3 patients died. Cardiovascular surgical patients of over 50 years or with angina pectorsi and ECG confirmed myocardiac ischemia should undergo coronary angiography routinely. If main coronary artery branches stenosis occupied over 50%, CABG must be performed. During the operation revasculariztion should be made as full as possible to enhance myocardiac protection and reduce the ascending aortic cross-clamping time. PMID- 10678076 TI - [Isolation of human herpes virus 6 from peripheral blood of renal transplant recipients]. AB - One strain of the viruses was isolated from preipheral blood lymphocytes (PBL) of a renal transplant recipient. PBL isolated from blood samples were cocultured with the PHA actived cord blood lymphocytes (CBL). Two of twelve recipient's samples found cytopathic effect after 10 to 14 days. Examination of ultrathin sections of the virus infected cells by electron microscope showed herpes-like virus particles. Detection of indirect immunofluorescences with McAbs against HHV 6 was positive in the infected cells. PMID- 10678077 TI - [Diagnosis and treatment of postrenal acute renal failure]. AB - 52 cases of postrenal acute renal failure (ARF) from 1985 to 1995 were studied. 50 cases underwent emergency operation, and 2 were drained with ureter intubation by cystoscope. 37 cases (71.2%) were cured, 14 (26.9%) were improved, and 1 (1.9%) died. Oliguria, anuria and progressive increase of blood urea nitrogen and serum creatinine are the main points of diagnosis. Renal percussive pain is the important sign. B-ultrasonography examination is the first choice and often indicate the increase of the volume of kidney and mild hydronephrosis. Obstruction should be removed as quickly as possible, infection should be prevented and treated to protect renal function. The way of treatment should be adopted according to the variant causes and conditions of disease. The etiology, clinical findings, diagnosis, operating methods and cautions were discussed. PMID- 10678078 TI - [The role of PMN CD11b/CD18 on the increasing PMN adhesion to endothelial cells induced by severe burn injury]. AB - The process of leukocyte adhesion was mediated by the intercellular adhesion molecular. In this study, both the influence of burn serum on the expression of PMN CD11b/CD18 and the role of CD11b/CD18 on the burn serum stimulated PMN adhesion to endothelial cells (EC) were investigated. The expression of CD11b/CD18 on PMN incubated with burn serum for 2 hours was increased obviously. So was PMN adhesion to EC. Monoclone antibody to PMN CD11b/CD18 could reduce the normal PMN adhesion to EC by half and block the increasing PMN adhesion induced by burn serum. These indicated that PMN CD11b/CD18 may be the main adhesion molecular participating in the increasing PMN adhesion to EC due to severe burn. PMID- 10678079 TI - [Preparation and clinical application of blocking glue for cancerous serosa]. AB - The blocking glue of cancerous serosa (F-TH glue) is unharmful to human body. It has characteristics of shorter solidification time to become membrane, good elasticity and close adherence to tissue, mainly used for covering cancerous serosa in operation to prevent dropping of cancer cells. In this group, F-TH glue was used in 200 cases of gastrointestinal cancer, in which 75 had positive results of imprinting slice of cancerous serosa, and all became negative. It was confirmed that the glue adhered to cancerous serosa firmly and formed intact protective membrane. No rhagades and dropping of glue were seem by naked eye magnifying glass and electron microscope. The membrane of glue forbid cancer cells not penetrated. It was an excellent glue to block the cancerous serosa. PMID- 10678080 TI - [Treatment of liver or renal cyst by percutaneous injection of TH glue into the cyst cavities]. AB - From April 1995 to May 1996, 32 cases of liver or renal cyts were detected by B ultra sound. Among them, TH glue was injected into the cyst to make it coagulated and occluded in 4 cases of liver cyst and 8 cases of renal cyst after intracapsular fluid was abstracted. The curative effect was good. The curative rate of single injection was 83.3%. The period of treatment and the curative rate were obviously different from the past methods. The present method is easy, quick, small wounded, few complicated and no toxic and side effects. PMID- 10678081 TI - Dynamic digital fluorescence ratio imaging of cell calcium in vascular endothelial cells. AB - AIM: To study the spatial and temporal distribution of intracellular Ca2+ concentration in cultured bovine pulmonary artery endothelial (BPAE) cells. METHODS: Cultured BPAE cells were loaded with Fura-2 and observed under an inverted microscope coupled to a microfluorimeter, which enables pixel-to-pixel ratio imaging of the BPAE cells in real time. RESULTS: Addition of Ca2+ 1-2 mmol.L-1 to BPAE cells, which were exposed to Ca(2+)-free medium containing egtazic acid, resulted in a transient elevation of cytosolic Ca2+ concentration, which rapidly returned to the resting level. Biphasic elevation (a larger transient phase followed by a smaller sustained phase) of intracellular Ca2+ concentration was observed upon the addition of ATP (via activation of surface membrane receptor). 4-Chloro-3-ethyl phenol (CEP; an activator of Ca(2+)-induced Ca2+ channels) potently induced elevation of Ca2+ level. Cyclopiazonic acid (CPA; an inhibitor of endoplasmic reticulum Ca(2+)-ATPase pump) offered a more sustained elevation of Ca2+. In most cases, the highest level of Ca2+ elevation was observed around the cell peripheries, sometimes at rest and particularly upon stimulation. Ca2+ elevation associated with nuclear complex seemed to be higher compared to that in the cytosolic compartment. CONCLUSION: Changes of cell Ca2+ upon stimulation by various agents that acted at different intracellular sites were found to be temporarily and spatially heterogenous among BPAE cells. At the single cell level, Ca2+ elevation seemed to occur initially near the peripheral region followed by the nuclear region. This study raised the possibility that nuclear Ca2+ and cytosolic Ca2+ might be regulated independently in BPAE cells. PMID- 10678082 TI - 6,7-dimethoxycoumarin attenuated cisplatin-induced DNA interstrand crosslink and DNA-protein crosslink in primary cultured rabbit kidney proximal tubular cells. AB - AIM: To study the mechanism of cisplatin interaction with DNA, and the attenuating effects of 6,7-dimethoxycoumarin (DMOC) on crosslink. METHODS: Primary cultured rabbit kidney proximal tubular cells (PTC) were established. DNA interstrand crosslink was assayed with ethidium bromide binding and DNA-protein crosslink with 125I-postlabelling. PTC were incubated with cisplatin for 24 h. DMOC was preincubated with PTC for 24 h, and cisplatin (26 mumol.L-1) was added into culture and incubated for another 24 h. RESULTS: Cisplatin induced formation of DNA interstrand crosslink (13, 26, 52, and 78 mumol.L-1) and DNA-protein crosslink (26, 52, and 78 mumol.L-1) (P < 0.01). DNA interstrand crosslink in DMOC (0.4, 4, and 8 mg.L-1) and DNA-protein crosslink in DMOC (4, 8 mg.L-1) were less than those in cisplatin group (26 mumol.L-1), respectively (P < 0.01). CONCLUSION: The mechanisms of cisplatin interaction with DNA in PTC were DNA interstrand crosslink and DNA-protein crosslink, and DMOC attenuated these effects in vitro. PMID- 10678083 TI - Cytosolic-Ca2+ and coxsackievirus B3-induced apoptosis in cultured cardiomyocytes of rats. AB - AIM: To explore the role of cytosolic free calcium ([Ca2+]i) in apoptosis induced by coxsackievirus B3 (CVB3) in cultured cardiomyocytes of rats. METHODS: Primary cultured cardiomyocyte was prepared from Wistar rats ages 2-3 d. The apoptosis in cardiomyocyte was determined by terminated deoxynucleotide transferase directed d UTP nick and end labeling (TUNEL) method, and the apoptosis was observed under a transmission electron microscope. [Ca2+]i in single cardiomyocyte loaded with Fluo 3-AM was measured by confocal microsorope. RESULTS: (1) The concentration of CVB3 in the medium reached the peak at 24 h after CVB3 infection. (2) The apoptotic cells were not found in CVB3-infected cardiomyocyte in first 10 h, but amounted to 5% at 17 h, 60% at 24 h, and 90% at 36 h. (3) The peak value of [Ca2+]i elevation reached at 17 h after CVB3 infection (P < 0.01). (4) The characteristics of apoptosis was also seen by transmission electron microscope. CONCLUSION: CVB3 induced the apoptosis in cultured cardiomyocyte, and [Ca2+]i mobilization was involved in the signal transduction process in apoptosis cells, and played an important role especially in the early stage of apoptosis induced by CVB3. PMID- 10678084 TI - Effects of anordrin, droloxifene, nomegestrol, and mifepristone on cultured rat luteal cell apoptosis. AB - AIM: To study the effect of four kinds of antifertility agents anordrin(Ano), droloxifene(Dro), nomegestrol (Nom), and mifepristone (Mif) on luteal cell apoptosis. METHODS: Cultured rat luteal cells were incubated with different agents. HE stain was used to observe morphological changes. Extracted DNA was electrophoresed on agarose gel. Apoptotic cells were quantitated by flow cytometry. RESULTS: All 4 drugs reduced cell viability. Dro induced apoptosis while the other 3 drugs induced necrosis. Typical DNA ladders were observed after cells were incubated with Dro and there were 15.4%, 75.4%, or 90.5% apoptotic cells after treatment with Dro 1.25, 2.5, or 3.75 mg.L-1, respectively. CONCLUSION: Dro induced apoptosis while Ano, Nom, and Mif induced necrosis in cultured rat luteal cells. PMID- 10678085 TI - Systemic anti-inflammation by synthetic interleukin-1 blockers. AB - AIM: To study the systemic anti-inflammatory actions of interleukin-1 (IL-1) blockers, OB-101 and OB-186. METHODS: Prevention of palm swelling induced by carrageenin injection was used as an animal model of systemic anti-inflammation efficacy. RESULTS: Both OB-101 and OB-186 (10-30 mg.kg-1) were approximately 10 30-fold more potent than aspirin (300 mg.kg-1) to inhibit carrageenin-induced systemic inflammation. The LD50 of OB-101 and OB-186 were at least 20 g.kg-1 i.g., indicating that they were extremely safe agents with a therapeutic index (LD50/ED50) of at least 2000. CONCLUSION: These IL-1 blockers are extremely safe systemically and are useable for the treatment of systemic inflammation such as rheumatoid arthritis. PMID- 10678086 TI - Protective effect of melatonin on injuried cerebral neurons is associated with bcl-2 protein over-expression. AB - AIM: To study the protective effect of melatonin against neuronal injury and the possible roles of alteration in the expression of bcl-2 and bax following brain ischemia. METHODS: Brain ischemia was induced by left middle cerebral artery occlusion (MCAO) for 60 min in rats. Brain damage was evaluated by the infarct area and the neuronal cell counting. The expression of bcl-2 and bax was analyzed by immunohistochemical method. RESULTS: Melatonin decreased the infarct area and prevented the neuronal death after 24-h reperfusion following 1-h MCAO. Melatonin given before the ischemia enhanced the expression of bcl-2 in the penumbra area and had no significant effect on the expression of bax. CONCLUSION: Melatonin effectively attenuated ischemic brain injury and increased the expression of neuronal bcl-2 in the ischemic brain, indicating that the protective effect of melatonin was associated with up-regulation of bcl-2 in ischemia-induced neuronal death. PMID- 10678087 TI - Effects of calcium channel blockers on calcium release-activated calcium currents in rat hepatocytes. AB - AIM: To study the influences of calcium channel blockers on calcium release activated calcium currents (ICRAC) in rat hepatocytes. METHODS: Whole-cell patch clamp technique was used. RESULTS: The peak amplitude of ICRAC was -0.41 nA +/- 0.09 nA (n = 15), its reversal potential was about 0 mV. Verapamil (Ver), diltiazem (Dil), and nifedipine (Nif) decreased ICRAC strikingly, without affecting its reversal potential. The inhibitory rate of Ver 5 mumol.L-1 was 40% +/- 12% (n = 3), Ver 50 mumol.L-1 reduced the peak amplitude of ICRAC from -0.49 nA +/- 0.12 nA to -0.20 nA +/- 0.09 nA (P < 0.01 vs control, n = 5). The inhibitory rate was 57% +/- 15%. Dil 50 mumol.L-1 and Nif reduced ICRAC from 0.43 nA +/- 0.10 nA to -0.29 nA +/- 0.07 nA (P < 0.01 vs control, n = 5), from 0.32 nA +/- 0.08 nA to -0.27 nA +/- 0.08 nA (P < 0.01 vs control, n = 5). The inhibitory rate was 31% +/- 11%, 19% +/- 7%, respectively. The amplitude of ICRAC was dependent on extracellular Ca2+ concentration. The peak amplitude of ICRAC was -0.21 nA +/- 0.08 nA (n = 3) in Tyrode's solution with Ca2+ 1.8 mmol.L-1 (P < 0.01 vs the peak amplitude of ICRAC in external solution with Ca2+ 10 mmol.L-1). CONCLUSION: The three calcium antagonists inhibited ICRAC effectively and protected hepatocytes from calcium overload via the inhibition of ICRAC. PMID- 10678088 TI - Devazepide reversed effect of sincalide against morphine on rat jejunal activities. AB - AIM: To study the antagonism of sincalide to the effect of morphine and its mechanism. METHODS: The electrophysiologic and mechanic activities of rat jejunum in vitro were recorded. RESULTS: Acetylcholine (ACh, 150 nmol.L-1) increased the spike potential amplitude (SPA) and the number (SPN) of rat jejunum in vitro, followed by an increase of jejunal contraction amplitudes (CA), showing a positive correlation. Morphine 330 nmol.L-1 inhibited the potentiation of ACh, showing a negative correlation. Sincalide 0.7 nmol.L-1 antagonized the effects of morphine, i.e., the SPA and SPN were increased again, followed by an increase of CA. CCK-A receptor antagonist devazepide (10 nmol.L-1) reversed the antagonism of sincalide to the effect of morphine. CONCLUSION: Sincalide antagonized the effect of morphine which inhibited the potentiation of ACh on jejunal activities in vitro. The antagonistic effect of sincalide on morphine was mainly mediated by CCK-A receptor. PMID- 10678090 TI - Quercetin decreased heart rate and cardiomyocyte Ca2+ oscillation frequency in rats and prevented cardiac hypertrophy in mice. AB - AIM: To study the effects of quercetin (Que) on myocardial excitation-contraction coupling and cardiac remodeling. METHODS: Left ventricles and femoral arteries of rats were cannulated for hemodynamic recording. Mouse cardiac hypertrophy was induced by abdominal aortic coarctation (AAC). Cultured myocardial cells in neonatal rats were loaded with Fura 2-AM. The intracellular calcium ([Ca2+]i) and spontaneous [Ca2+]i oscillations ([Ca2+]i-SO) were tested by AR-CM-MIC cation measurement system. RESULTS: Que 3 or 25 mg.kg-1 i.v. in rats decreased heart rate from (420 +/- 19) to (390 +/- 15) and (314 +/- 18) beat.min-1, respectively, companied with very modest changes in both left ventricular pressures (LVP) and its differential dpLV/dtmax. Que 10, 50, 250 mumol.L-1 concentration-dependently slowed the frequency of [Ca2+]i-SO in cultured myocardial cells from (26 +/- 4) to (25 +/- 3), (18 +/- 4), and (12 +/- 3) time.min-1, respectively, but did not change their resting [Ca2+]i or amplitudes of [Ca2+]i-SO. Similarly, the increases in frequency of [Ca2+]i-SO caused by either isoproterenol (Iso) or ouabain (Oua) were prevented by Que 100 mumol.L-1, while the simultaneous increases in amplitude of [Ca2+]i-SO remained. Besides, [Ca2+]i rises excited by angiotensin II (Ang II) but not high [K+]o were prevented by Que 100 mumol.L-1. Daily administration of Que 120 mg.kg-1 i.g. for 5 d markedly prevented the cardiac hypertrophy in AAC mice, without effects on the ventricular mass to body weight ratio (VM/BW) in sham-operated mice. CONCLUSION: Que decreased myocardial [Ca2+]i-oscillation frequency and prevented cardiac remodeling, but had no direct effect on cardiac excitation-contraction coupling. PMID- 10678089 TI - Antagonistic effects of Ginkgo biloba extract on adhesion of monocytes and neutrophils to cultured cerebral microvascular endothelial cells. AB - AIM: To study the action of Ginkgo biloba extract (GbE) on tumor necrosis factor (TNF-alpha)-induced adhesion of monocytes (Mon) and neutrophils (Neu) to cultured cerebral microvascular endothelial cells. METHODS: TNF-alpha-induced endothelial adhesivity toward Mon and Neu was studied using bovine cerebral microvascular endothelial cells (BCMEC) in vitro. The number of Mon and Neu adhering to the BCMEC monolayers was determined by flow cytometry. RESULTS: Pretreatment of BCMEC with TNF-alpha increased Mon and Neu adhesion to BCMEC from 12.5% +/- 0.2% to 31.3% +/- 0.5% and from 13.8% +/- 0.4% to 32.1% +/- 0.5%, respectively. GbE (1 100 mg.L-1) inhibited the effect of TNF-alpha in a concentration-dependent manner. E-selectin mAb (1 mg.L-1) blocked Mon and Neu adhesion to BCMEC induced by TNF-alpha. CONCLUSION: The inhibition of GbE on Mon and Neu adhesion to BCMEC was mediated through the suppression of E-selection expression. PMID- 10678091 TI - Selective effects of alfuzosin and doxazosin with intraduodenal administration on urethral pressure of cats. AB - AIM: To observe the selective effects of alfuzosin (Alf) and doxazosin (Dox) on the urethral pressure by different administration routes. METHODS: The urethral pressure of the anesthetized cat was increased by electric stimulation of the hypogastric nerve. The different effects of Alf or Dox on the arterial blood pressure and urethral pressure between intraduodenal administration (i.d.) and intravenous infusion (i.v.) were compared. RESULTS: When the hypogastric nerve was stimulated by electric stimulation (10 Hz, 25 V), the ratios of ED20(BP)/ED50(UP) i.d. to ED20(BP)/ED50(UP) i.v. were 10.9:4.3 for Alf, and 3.1:2.1 for Dox. The reduction in urethral pressure induced by i.d. Alf was greater than that by i.v. Alf. Dox did not show any difference in its effects by 2 administration routes. CONCLUSION: Intraduodenal administration of Alf, but not Dox, selectively decreased the urethral pressure elevated by electric stimulation. The uroselectivity of i.d. Alf was not due to the species difference in its bioavailability and biotransformation. PMID- 10678092 TI - Circumvention of tumor multidrug resistance by a new annonaceous acetogenin: atemoyacin-B. AB - AIM: To explore the effect of atemoyacin-B (Ate) on overcoming multidrug resistance (MDR). METHODS: Bullatacin (Bul) was used as a positive control. Cytotoxic effects of Bul and Ate were studied with cell culture of human MDR breast adenocarcinoma cells, MCF-7/Dox and human KBv200 cells, and their parental sensitive cell lines MCF-7 and KB. Cytotoxicity was determined by tetrazolium (MTT) assay. The function of P-glycoprotein (P-gp) was examined by Fura 2-AM assay. Cellular accumulation of doxorubicin (Dox) was determined by fluorescence spectrophotometry. Apoptosis was measured by flow cytometry. RESULTS: IC50 of Ate for MCF-7/Dox, MCF-7, KBv200, and KB cells were 122, 120, 1.34, and 1.27 nmol.L 1, respectively. IC50 of Bul for MCF-7/Dox, MCF-7, KBv200, and KB cells were 0.60, 0.59, 0.04, and 0.04 nmol.L-1, respectively. The cytotoxicities of Bul and Ate to MDR cells were similar to those to parental sensitive cells. Bul and Ate markedly increased cellular Fura-2 and Dox accumulation in MCF-7/Dox cells, but not in MCF-7 cells. The rates of apoptosis in MDR cells were similar to those in sensitive cells induced by Ate. CONCLUSION: There was no cross-resistance of P-gp positive MCF-7/Dox and KBv200 cell lines to Bul and Ate as compared with their sensitive P-gp negative MCF-7 and KB cell lines. The mechanism of the circumvention of MDR was associated with the decrease of P-gp function and the increase of cellular drug accumulation in MDR cells. PMID- 10678093 TI - Gene therapy for human hepatocellular carcinoma with cytosine deaminase gene and prodrug flucytosine. AB - AIM: To investigate the antitumor effects of cytosine deaminase (CD) gene in combination with prodrug flucytosine (Flu, 5-fluorocytosine) on human hepatocellular carcinoma. METHODS: CD gene was transduced into human hepatocellular carcinoma cell line SMMC7721 with retroviral method and the cytotoxicity of Flu on the tumor cells was assayed in vitro with clonogenic techniques. The xenograft tumor model in nude mice was used to study in vivo therapeutic effects of CD gene/Flu system against human hepatocellular carcinoma. RESULTS: CD gene/Flu system had significant antitumor activities on human hepatocellular carcinoma cells in vitro and in nude mice. The antitumor activities of Flu 500 mg.kg-1 on hepatocellular carcinoma xenografts in nude mice were more potent than those of 5-fluouracil 10 mg.kg-1. CD gene/Flu system possessed bystander killing effects on hepatocellular carcinoma xenografts in nude mice. CONCLUSION: The experiment demonstrates the potential value of the CD gene/Flu system in the treatment of human hepatocellular carcinoma. PMID- 10678094 TI - Apoptosis and necrosis induced by sulfur mustard in Hela cells. AB - AIM: To study the apoptotic effect of sulfur mustard (SM) on Hela cells. METHODS: Exponentially growing Hela cells were treated with SM at various concentrations for 3 h, then apoptosis was examined by electron-microscope, DNA gel electrophoresis, and flow cytometry. RESULTS: SM 1 mumol.L-1 arrested cell growth. After treatment with SM 10-100 mumol.L-1, cells were mainly blocked at G1 phase with apoptosis. Agarose gel electrophoresis of DNA from cells treated with SM revealed "DNA Ladder." About 33% of the Hela cells showed apoptosis 12 h after 3-h treatment with SM 100 mumol.L-1 as determined by flow cytometry and the S phase cells were more susceptible. However, SM 1000 mumol.L-1 caused marked necrosis in Hela cells. CONCLUSION: SM caused 2 distinct forms of cell death, apoptosis or necrosis, in Hela cells in a concentration-dependent manner. PMID- 10678095 TI - Ro 31-8220 inhibits release of interleukin-1 and interleukin-6 from mouse peritoneal macrophages induced by fibrin fibrinogen degradation products. AB - AIM: To study the effect of fibrin fibrinogen degradation products (FFDP) on release of interleukin-1 (IL-1) and interleukin-6 (IL-6) from mouse peritoneal macrophages, and the effect of a new selectively potent protein kinase C inhibitor Ro 31-8220 (Ro). METHODS: IL-1 and IL-6 activities were measured by thymocyte proliferation assay and B9 cell proliferation methyl thiazolyl tetrazolium (MTT) colorimetric method, respectively. RESULTS: Ro 0.01-1 mumol.L-1 obviously inhibited FFDP-induced release of IL-1 and IL-6 from mouse peritoneal macrophages. CONCLUSION: Ro exerted inhibitory effects on FFDP-induced release of IL-1 and IL-6 in vitro. PMID- 10678096 TI - Effects of opioid receptor agonists on cAMP second messenger system. AB - AIM: To study the mechanism underlying the difference in physical dependence potential of morphine (Mor), methadone (Met), buprenorphine (Bup), etorphine (Eto), and dihydroetorphine (DHE). METHODS: Adenylate cyclase of NG108-15 cells were used for studying the effects of different opiates on cAMP second messenger system. RESULTS: Bup, DHE, and Eto were distinct from Mor in naloxone precipitated rebound response of cAMP in NG108-15 cells chronically treated with these opiates. Naloxone given to NG108-15 cells treated with Mor for 24 h produced marked rebound response of adenylate cyclase. While no such rebound response was detected when the cells were treated with Bup, DHE, and Eto for 24 h. The naloxone-induced rebound response of cAMP in chronic Met-treated NG108-15 cells was also lower than that in chronic Mor-treated NG108-15 cells. Following a prolonged exposure to Bup, DHE, and Eto for 72 h, the naloxone-induced rebound response of cAMP in these cells was still markedly lower than that in Mor-treated cells. The substitution of Mor with Bup, Met, DHE, and Eto inhibited naloxone induced rebound response of cAMP in chronic Mor-treated NG108-15 cells. CONCLUSION: There were distinct differences among these opiates in regulating cAMP second messenger system, which was related to their physical dependence potential. PMID- 10678097 TI - Anti-inflammatory and nitric oxide-inhibiting properties of granulocyte colony stimulating factor. AB - A proposed scheme between the possible interactions of pro- and anti-inflammatory cytokines, NO and G-CSF during severe inflammation/infection is presented. Taken together, these data indicate that G-CSF exhibits anti-inflammatory properties which may prove to be beneficial in situations associated with an increased activity of the cellular immune system. Since the suppressive effects of G-CSF on the production of pro-inflammatory mediators like TNF-alpha and nitric oxide are most likely neither cell type nor tissue specific, it is conceivable that NO release induced by pro-inflammatory mediators can be reduced by G-CSF in various organ systems and in different forms of shock. In this context, G-CSF might represent a counterregulatory mechanism directed against a downstream oriented inflammatory response to infection. Therefore, the investigation of G-CSF in the prophylaxis of nonneutropenic infections, sepsis, and other severe inflammatory disorders seems reasonable. PMID- 10678098 TI - Vascular effects of estrogens: rapid actions, novel mechanisms, and potential therapeutic implications. AB - Although estrogen-dependent effects on the vasculature were first observed more than a century ago, many of the mechanisms by which estrogens interact with the vascular wall have been identified only in the past 15 years. Estrogens bind to vascular estrogen receptors (ER), including the ER alpha, the novel ER beta as well as to membrane-bound receptors. Estrogens have direct effects in human coronary and internal mammary arteries by inducing rapid, endothelium-independent relaxation, enhancement of endothelial function and inhibition of vasoconstriction by vasoactive agonists. Furthermore, estrogens contribute to vascular homeostasis through modulation of gene expression, changes in membrane potentials, as well as expression and function of receptors. In addition, estrogens interfere with the activity of vasoactive peptides and vascular enzymes and act as natural antioxidants. Some of these effects have also been observed for phyto-estrogens, which are important dietary components in Asian countries. In the vasculature, the sum of these actions of estrogens results in vasodilatation and inhibition of vascular cell growth. Accordingly, estrogens have been shown to improve vascular function of animals and humans and to inhibit the response to injury after balloon angioplasty and the progression of atherosclerosis. Prospective clinical studies are ongoing to determine whether replacement therapy with estrogen or derivatives provides an alternative to lower cardiovascular mortality in postmenopausal women. PMID- 10678099 TI - Comparison of effects of surfactant and inhaled nitric oxide in rabbits with surfactant-depleted respiratory failure. AB - AIM: To compare effects of pulmonary surfactant and inhaled nitric oxide (iNO) in improvement of survival and blood oxygenation in ventilated rabbits with acute hypoxic respiratory failure induced by repeated bronchoalveolar lavage (BAL). METHODS: After BAL all the rabbits had more than 50% reduction of dynamic lung compliance (Cdya), 50% increment of resistance of respiratory system (Rrs), and an increase of mean oxygenation index (OI) from 1 to 22. The rabbits were then randomly allocated to groups receiving (1) mechanical ventilation only (Control), (2) iNO 0.8 mumol.L-1 (20 ppm) (NO), (3) intratracheal bolus surfactant phospholipids at 100 mg.kg-1 (Surf), and (4) combined surfactant at 100 mg.kg-1 with inhaled NO at 0.8 mumol.L-1 (Surf + NO). All the rabbits were ventilated with standardized tidal volume (8-10 mL.kg-1) for another 8 h or until early death. RESULTS: The rabbits in both control and NO groups had the lowest survival rate, deterioration of lung mechanics and OI, whereas those in the Surf and Surf + NO groups had modestly improved Cdyn, Rrs, and OI. Only rabbits in the Surf + NO group had significantly improved survival rate and alveolar expansion. CONCLUSION: Surfactant with or without iNO is more effective compared to the control and iNO groups in rabbit, suggesting that iNO is not effective unless a method to recruit alveoli is applied. PMID- 10678100 TI - dl-3-n-butylphthalide attenuates reperfusion-induced blood-brain barrier damage after focal cerebral ischemia in rats. AB - AIM: To study the protective effect of dl-3-n-butylphthalide (NBP) on blood-brain barrier (BBB) damage induced by reperfusion following focal cerebral ischemia. METHODS: Focal cerebral ischemia in rats was performed by inserting a nylon suture into intracranial segment of internal carotid artery to block the origin of middle cerebral artery and reperfusion by withdrawing the nylon suture. Permeability of BBB was determined by extravasation of the protein-bound Evans blue dye to cerebral cortex and further evaluated by immunohistochemical or electronmicroscopic method. RESULTS: Reperfusion for 3 h following focal cerebral ischemia for 3 h produced BBB damage which exhibited the increase in extravasation in cerebral cortex, elevation of the expression of immunoglobulin (IgG), and pore formation in endothelial cell membrane of capillary in cerebral cortex. NBP (5-20 mg.kg-1) decreased the extravasation in a dose-dependent manner. The expression of IgG in cerebral cortex was decreased and the ultrastructure damage of capillaries was alleviated after treatment with NBP. NBP 20 mg.kg-1 also alleviated brain edema caused by 3-h reperfusion following 3-h middle cerebral artery occlusion (MCAO). CONCLUSION: NBP has protective effect on BBB damage induced by reperfusion after MCAO. PMID- 10678101 TI - Antagonistic effects of shikimic acid against focal cerebral ischemia injury in rats subjected to middle cerebral artery thrombosis. AB - AIM: To study the effects of shikimic acid (SA) on focal cerebral ischemic injury after middle cerebral artery thrombosis (MCAT). METHODS: Thrombosis was induced by FeCl3 in middle cerebral artery of rats. The influences of SA on neurologic deficit (ND), infarct size (IS), brain edema, and cerebral blood flow (CBF) in ischemic region were observed. RESULTS: SA 25 and 50 mg.kg-1 i.p. for 3 d before MCAT attenuated ND, and reduced IS by 51% and 42%; and decreased brain water content from 80.7% to 79.8% and 79.9%; and increased CBF after ischemia from 50.2% of the preischemic level to 75.5% and 73.3%, respectively. In pathologic examination, there was much less thrombosis in MCA in the rat with the pretreatment by SA 25 mg.kg-1. The extent of brain ischemia was much less than that of control. CONCLUSIONS: SA reduced focal cerebral ischemic injury induced by middle cerebral artery thrombosis. PMID- 10678102 TI - Effects of berbamine on ATP-induced [Ca2+]i mobilization in cultured vascular smooth muscle cells and cardiomyocytes. AB - AIM: To study the effects of berbamine (Ber) on [Ca2+]i homeostasis induced by adenosine triphosphate (ATP) in vascular smooth muscle cells (VSMC) of rabbits and cardiomyocytes of rats. METHODS: Both cell types were cultured and loaded with Fura 3-AM. [Ca2+]i was measured by fluorescent intensity (FI) in each cell with confocal microscopy. RESULTS: (1) ATP 30 mumol.L-1 elevated [Ca2+]i in VSMC and cardiomyocytes, FI values reached 660 +/- 258 and 1058 +/- 252 from 250 +/- 84 and 218 +/- 76 at 19 s +/- 5 s and 11.8 s +/- 2.4 s, but FI in nucleus was not changed in VSMC. (2) Ber 30 mumol.L-1 did not affect the resting FI in both cell types, but prolonged the time to peak (P < 0.01) and reduced the FI elevated by ATP (P < 0.01), but not completely inhibited even at 100 mumol.L-1. (3) In D Hanks' solution or in the presence of egtazic acid (EGTA) 3 mmol.L-1, the inhibitory effect of Ber was not seen (P > 0.05). (4) All effects of Ber on ATP induced [Ca2+]i mobilization were similar to those of Ver 10 mumol.L-1. CONCLUSION: In VSMC and cardiomyocytes, ATP-induced CA2+ influx was inhibited by Ber and Ver, while the Ca2+ release was not. PMID- 10678103 TI - Antiviral effects of rhIFN-alpha 1 against seven influenza viruses. AB - AIM: To study the antiviral effects of rhIFN-alpha 1 (Chinese silkworm gene recombinant interferon alpha 1) on 7 influenza viruses in MDCK cells and in mouse pneumonia caused by PR8 virus. METHODS: 100TCID50 virus (H1N1, H2N2, H3N3, type B, type C, clinical A1, and clinical B) were inoculated into MDCK cells, PR8 viruses were dropped nasally in mice, the antiviral effects of rhIFN-alpha 1 were observed. RESULTS: The minimal effective concentrations of rhIFN-alpha 1 against these 7 influenza viruses were 12.5, 25, 50, 25, 12.5, 25, and 12.5 kU.L-1, respectively. The infectious therapeutic indices of rhIFN-alpha 1 to these viruses in MDCK cells were 8 x 10(3), 4 x 10(3), 2 x 10(3), 4 x 10(3), 8 x 10(3), 4 x 10(3), and 8 x 10(3), respectively. The inhibitory indices of rhIFN-alpha 1 to the 7 influenza viruses in MDCK cells were 3.6, 4.7, 3.5, 3.3, 3.9, 4.6, and 3.5, respectively. The rhIFN-alpha 1 inhibited the intracellular replication of influenza viruses effectively, but did not kill viruses directly. The rhIFN-alpha 1 prolonged the life span of mice infected with pneumonia by influenza virus A strain PR8 to 94.2%-132.7%. It inhibited the inflammation and hyperplasia of interstitial fibers, and decreased the virus titer. The inhibitory rates of rhIFN alpha 1 to pulmonary-indice were 14.8%-37.4%. CONCLUSION: rhIFN-alpha 1 inhibited the proliferation of influenza virus and improved the symptom of mouse pneumonia caused by influenza virus. PMID- 10678104 TI - Supraspinal D2 receptor involved in antinociception induced by l tetrahydropalmatine. AB - AIM: To study the role of dopamine (DA) receptors in l-tetrahydropalmatine (l THP)-induced antinociception. METHODS: The intraperitoneal (i.p.) and intrathecal (ith) injections were used to give the drugs. The tail-flick test was used to assess the nociceptive threshold of rats. RESULTS: By i.p. injection, l-THP (10, 20, 40 mg.kg-1) as well as D2 receptor antagonist spiperone (1, 2, 3 mg.kg-1) produced dose-dependent antinociceptive effects on the nociception of rats, while D2 receptor agonist quinpirole, D1 receptor agonist SKF38393, and D1 receptor antagonist Sch-23390 showed no antinociception. Moreover, l-THP- or spiperone induced antinociception was markedly attenuated by quinpirole (2, 3 mg.kg-1) but not SKF38393 or naloxone. On the other hand, ith quinpirole (20, 30, 40 micrograms.kg-1) also induced a dose-dependent antinociception, while ith l-THP, spiperone, SKF38393, and Sch-23390 did not exhibit any antinociception. Furthermore, ith spiperone (20, 30, 40 micrograms.kg-1) but not Sch-23390 dose dependently antagonised the antinociception induced by quinpirole. l-THP (ith, 100, 200, 300 micrograms.kg-1) also dramatically attenuated the quinpirole induced antinociception with a dose-dependent relationship. CONCLUSION: Activating the spinal D2 receptor or blocking the supraspinal D2 receptor produces antinociception. D1 receptor might be not directly involved in the antinociception. l-THP (as a D2 antagonist) as well as spiperone produces antinociception via blocking the supraspinal D2 receptor. PMID- 10678105 TI - CYP2D6 phenotype determines pharmacokinetic variability of propafenone enantiomers in 16 HAN Chinese subjects. AB - AIM: To determine the role of the CYP2D6 phenotype in the metabolism of propafenone (Pro) enantiomers in 16 HAN Chinese subjects. METHODS: Seven very extensive metabolizers (VEM) and nine intermediate metabolizers (IM) were enrolled from a Chinese population whose phenotype had been previously assessed with a dextromethorphan metabolic phenotyping. The blood samples (0-15 h) were taken after oral administration of a single dose (400 mg) of rac-Pro hydrochloride. Enantiomeric concentrations of propafenone in plasma were measured by a reverse-phase HPLC with precolumn derivatization. RESULTS: S-Pro was less metabolized and had higher plasma concentrations than R-Pro in both CYP2D6 phenotypes. Besides, the T1/2 of R-Pro was longer than that of S-Pro in IM, but not in VEM. However, there were significant differences in the metabolism of Pro enantiomers between VEM and IM. The Cmax and AUC of both isomers in the IM group were higher than those in the VEM group (P < 0.01). The Cl of Pro enantiomers in IM group was only about half of that in VEM group [(67 +/- 17) vs (133 +/- 28) L.h-1 for S-Pro, (90 +/- 24) vs (200 +/- 87) L.h-1 for R-Pro, P < 0.01]. The S/R ratios of T1/2, Tmax, Cmax, Cl, and AUC were not significantly different (P > 0.05). CONCLUSION: CYP2D6 phenotype determines the pharmacokinetic variability of propafenone enantiomers and existence of IM may be relevant to diminished capacity of CYP2D6 enzyme in Chinese subjects. PMID- 10678106 TI - Subtypes of central nicotinic receptors involved in learning and memory. AB - AIM: To observe the effects of different subtypes of central nicotinic receptor on learning and memory. METHODS: Passive avoidance response, including step-down avoidance and step-through avoidance in mice and long-term potentiation (LTP) in rat hippocampal slices. RESULTS: Hexamethonium (C6) 7 micrograms/mouse and kappa bungarotoxin (kappa-BTX) 0.6 microgram/mouse inhibited the acquisition of avoidance conditioning in mice, and kappa-BTX yielded this effect with a dose response relationship. kappa-BTX 1 mumol.L-1 suppressed the induction of LTP (P < 0.05), but not normal synaptic transmission and maintenance of LTP in rat hippocampal slices. CONCLUSION: The subtypes of central nicotinic receptor sensitive to kappa-BTX play an important role in learning and memory. PMID- 10678107 TI - Effects of tetrandrine on changes of NMDA receptor channel in cortical neurons of rat induced by anoxia. AB - AIM: To study the effects of tetrandrine (Tet) on the changes of NMDA receptor channels in cortical neurons induced by anoxia. METHODS: Cell-attached configuration of patch-clamp techniques. Anoxia was produced by perfused cells with 95% N2 + 5% CO2 gassed bath solution. RESULTS: During anoxia, the open time constant (tau 2), open probability (Po) of 35-pS and 100-pS channels increased. Tet 7.5 mumol.L-1 reduced the Po of 35-pS and 100-pS channels, 15 and 30 mumol.L 1 inhibited open of 100-pS channel fully, and changed the open time constant of 35-pS from two to single exponential distribution. CONCLUSION: Tet inhibition of the open of NMDA receptor channels induced by anoxia was one of its protective mechanisms. PMID- 10678108 TI - L-pyroglutamic acid protects rat cortical neurons against sodium glutamate induced injury. AB - AIM: To evaluate the effects of L-pyroglutamic acid (L-PGA, L-5-oxo-2 pyrrolidinecaroxylic acid) on sodium glutamate-induced neurotoxicity in rat cortical neurons. METHODS: In primary cortical cultures from 16-d-old fetal rat, neuronal viability and contents of nitrite in the bathing medium after transient exposure to sodium glutamate (Glu) were measured; with Fura 2-AM as an intracellular calcium indicator, AR-CM-MIC cation measurement system was used to examine cytosolic free calcium ([Ca2+]i). RESULTS: L-PGA 10-80 mumol.L-1, inhibited Glu (500 mumol.L-1)-induced neuronal loss in a concentration-dependent manner with IC50 value of (41 +/- 9) mumol.L-1 (95% confidence limits: 30.3-54.7 mumol.L-1). L-PGA also attenuated Glu-induced NO release. L-PGA 1, 3, 10, 30, and 100 mumol.L-1 depressed Glu-caused [Ca2+]i elevation by 20.5%, 34.4%, 47.7%, 70.6%, and 80.4%, respectively. CONCLUSION: L-PGA protects cortical neurons against Glu-induced neurotoxity which may be related to inhibition of NO formation or suppression of the rise in [Ca2+]i. PMID- 10678109 TI - Intrathecal injection of corticotropin inhibited nitric-oxide synthase-positive neuron increase in rat spinal cord after formalin-induced hyperalgesia. AB - AIM: To study the effects of corticotropin (Cor) on formalin-induced hyperalgesia and the change of nitric-oxide synthase (NOS)-positive neurons in spinal dorsal horn in rats. METHODS: Measurement of pain intensity rating (PIR), NADPH-d histochemistry, and Fos immunohistochemistry were adopted. RESULTS: The increases of NOS-positive neurons, Fos, NOS/Fos double labelling neurons of the spinal dorsal horn and the PIR after formalin injection were markedly inhibited by intrathecal injecting (ith) Cor (0.5-1.5 U), which were obviously attenuated by L arginine (Arg, 5-15 nmol, ith), the substrate of NOS. CONCLUSION: Cor inhibits formalin-induced hyperalgesia by the decrease of NOS-positive neurons in the spinal dorsal horn of rats. PMID- 10678110 TI - Effects of pentoxifylline and protein kinase C inhibitor on phorbol ester-induced intercellular adhesion molecule-1 expression in brain microvascular endothelial cells. AB - AIM: To study the potential roles of protein kinase C (PKC) on expression of intercellular adhesion molecule-1 (ICAM-1) in rat brain microvascular endothelial cells (RBMEC). METHODS: ICAM-1 expression in RBMEC was measured by ELISA analyses. RESULTS: Phorbol ester (PMA) enhanced the expression of ICAM-1 in a concentration (10-100 nmol.L-1) and time (4-16 h)-dependent manner in RBMEC. Pentoxifylline (PTX) 1-100 mumol.L-1 and the PKC inhibitor H7 5-50 mumol.L-1 prevented PMA-induced stimulation of ICAM-1 expression. At PTX 100 mumol.L-1 and H7 50 mumol.L-1, they reached maximal inhibitory effects [ICAM-1 expression (A) from (0.410 +/- 0.014) to (0.175 +/- 0.022) and (0.182 +/- 0.013), respectively; P < 0.01]. CONCLUSION: Activation of PKC in RBMEC is associated with increased expression of ICAM-1 in RBMEC. PTX and H7 preincubation may inhibit PKC-induced up-regulation of ICAM-1. PMID- 10678111 TI - Hydrolysis of extracellular adenine nucleotides by cultured bovine endocardial endothelial cells. AB - AIM: To characterize the ATP diphosphohydrolase (apyrase) of bovine endocardial endothelial cells, and to compare ecto-adeninenucleotidase activity between bovine endocardial and aortic endothelial cells (BEEC and BAEC). METHODS: The nucleotide was analyzed by reversed phase HPLC and apyrase activity was assayed by inorganic phosphate release. RESULTS: Apyrase inhibitors, both NaN3 10 mmol.L 1 and NaF 20 mmol.L-1, inhibited BEEC apyrase activity by 51% and 38%, respectively. The inhibitor for Na+/K(+)-ATPase, ouabain, did not affect the enzyme activity. Edetic acid 5 mmol.L-1 completely inhibited the enzyme activity. H2O2 0.5 mmol.L-1 downregulated BEEC apyrase activity in a time-dependent manner. The apyrases activities in BAEC were higher than those in BEEC, while the ecto AMPase activity in BAEC was much weaker than that in BEEC. CONCLUSION: BEEC have NaN3- and NaF-sensitive, ouabain-insensitive apyrase activity. BEEC had high ecto AMPase activities, and low apyrases activities as compared with BAEC. PMID- 10678112 TI - Effect of artemether on phosphorylase, lactate dehydrogenase, adenosine triphosphatase, and glucosephosphate dehydrogenase of Schistosoma japonicum harbored in mice. AB - AIM: To study the effect of artemether (Art) on phosphorylase (PP), lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (G-6-PDH), and adenosine triphosphatase (ATPase) of S japonicum. METHODS: Mice infected with S. japonicum cercariae for 32-38 d were treated i.g. with Art 100-300 mg.kg-1 and killed 24-72 h after treatment for collection of schistosomes. The activities of PP, LDH, and G-6-PDH were measured by the formation of NADH or NADPH. The activity of ATPase was measured by the rate of release of inorganic phosphate (Pi) from ATP at 37 degrees C. RESULTS: After infected mice were treated i.g. with Art 300 mg.kg-1 for 24-48 h, the activities of total PP and PPa (active form) increased markedly in both male and female worms, while PPb (inactive form) showed no or only a slight increase. At 24-72 h after the above-mentioned mice were treated i.g. with Art 100-300 mg.kg-1, the inhibitory rates of LDH and G-6-PDH were 9%-59% (male) and 41%-75% (female) as well as 22%-42% (male) and 74%-89% (female), respectively. When Art 300 mg.kg-1 was given to infected mice for 24 h, only the activity of Mg(2+)-ATPase showed marked inhibition in both male and female worms. At 48 h, the Ca(2+)-ATPase, Mg(2+)-ATPase, and Na(+)-K(+)-ATPase were all inhibited, the inhibitory rates of 17% (male) and 19% (female), 32% (male) and 48% (female) as well as 29% (male) and 44% (female), respectively. CONCLUSION: In schistosomes, the increase in the activity of AMP-independent PPa induced by Art may enhance the decomposition of glycogen and the inhibition of LDH by Art could reduce the formation of lactate. Moreover, Art exerts a potent inhibition on the G-6-PDH activity of the female S japonicum. PMID- 10678113 TI - Schizontocidal effects of oral artesunate on Plasmodium berghei in mice and P knowlesi in monkeys. AB - AIM: To study the blood schizontocidal effect of oral artesunate on P berghei in mice and P knowlesi in monkey. METHODS: Effects of artesunate and chloroquine were detected with "4-day test" and "28-day test" on P berghei in mice and "7-day test" on P knowlesi in Macaca mudatta. RESULTS: The suppressive efficacy of oral artesunate was inferior to chloroquine on P berghei K173 strain but the time for 50% and 90% reduction and the time of clearance of parasitemia was 10-15 h shorter than that of chloroquine. Its curative effect on RC/K173 line was markedly superior to that of chloroquine. Moreover, artesunate showed no cross resistance with chloroquine, index of resistance I90 was only 1.4. At 31.6, 10.0, and 3.16 mg.kg-1, artesunate and chloroquine oral administrations cured P knowlesi in all monkeys. Recrudescence did not occur in 105 d. CONCLUSION: The study of effects of oral artesunate in P berghei/mice and P knowlesi/Macaca mulatta model provided a useful index for clinical trial. PMID- 10678114 TI - Cytokines enhance nitric oxide production from human BT325 astrocytoma cells. AB - AIM: To study the effects of lipopolysaccharides (LPS) and pre-inflammatory cytokines on nitric oxide (NO) production from cultured astrocytes. METHODS: Nitrites in supernatants were measured with Griess assay. RESULTS: The NO production from cultured human BT325 astrocytoma cells started when cultured for 4 h, reached the peak concentration (15.0 mumol.L-1) at 12 h, maintained a high level (15.0-17.5 mumol.L-1) up to 72 h, and was enhanced by LPS 1 mg.L-1, interferon-gamma (IFN-gamma) 100 kU.L-1, tumor necrosis factor-alpha (TNF-alpha) 100 kU.L-1, interleukin (IL)-1 100 kU.L-1, or IL-2 100 kU.L-1. The enhancements of TNF-alpha, IL-1, IL-2, or the mixture of the above four cytokines were higher. CONCLUSION: Stimulants and pre-inflammatory cytokines enhance astrocytes producing NO. PMID- 10678115 TI - Effect of recombinant human basic fibroblast growth factor on stomach ulcers in rats and mice. AB - AIM: To study the curative effects of recombinant human basic fibroblast growth factor (rh-bFGF) on gastric ulcer healing. METHODS: Pylorus ligated, water immersion stress-induced, reserpine-induced, and acetic acid-induced gastric ulcers in rats or mice were prepared. Morphometric analyses on ulcer were performed using microscope and true color image analysis system. The DNA and RNA contents were measured by diphenylamine method and orcinol method, respectively. RESULTS: In acetic acid-induced gastric ulcers in rats, rh-bFGF 2.5-40 kU.kg-1 i.g. accelerated the chronic ulcer healing with a bell-shaped dose-effect curve and the best dosage was 10 kU.kg-1. The regenerated gland epithelium width, density of capillaries in granulation tissue, and collagen content in scar tissues obviously increased in rh-bFGF-treated groups. Simultaneously, rh-bFGF promoted the differentiation and maturation of regenerated glands around ulcers. rh-bFGF 2-4 kU.kg-1 s.c. also increased the synthesis of RNA in ulcer tissues. In acute gastric ulcers, rh-bFGF i.g. was only effective on pylorus ligated ulcers, but showed no effect on total acid output and pepsin activity in gastric juice of rats. CONCLUSION: rh-bFGF promoted the gastric ulcer healing and improved the quality of gastric ulcer healing. PMID- 10678116 TI - Pharmacological regulation of striatal gene expression by metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGluR) are densely expressed by striatal medium spiny neurons. Activation of mGluR in this brain region alters local transmitter release and behaviors of experimental animals. In particular, mGluR regulate transcription factor and neuropeptide gene expression in striatal neurons through their connections with multiple intracellular effectors. This prominent involvement of mGluR in overall cellular activity is pivotal for the development of neuronal plasticity underlying long-term adaptive changes in cellular physiology related to a variety of neurologic disorders. Accumulating evidence demonstrates that the subtypes of mGluR have distinct effects on gene expression: group I subtypes facilitating, and group II/III subtypes inhibiting, gene expression. Thus, the mGluR can be considered as promising targets in the development of novel therapeutic drugs that can relieve neurologic disorders resulting from dysfunction of the striatum. PMID- 10678117 TI - Correlation between cytochrome P-450 CYP2D6 (CYP2D6) genotype and phenotype. AB - AIM: To study the correlation between CYP2D6 genotype and its phenotype. METHODS: CYP2D6 genotyping was made by detecting CYP2D6*3, *4, *6, and *7 alleles with an allele-specific polymerase chain reaction procedure. RESULTS: The CYP2D6 genotypes were well correlated with its phenotypes in all 125 extensive metabolizers and in 43 poor metabolizers. Extensive metabolizers had at least one wildtype CYP2D6 gene and the genotypes were *1/*1, *1/*3, and *1/*4. Poor metabolizers were found to be homozygous mutants of CYP2D6 gene and the genotypes were *3/*4, *4/*4, *3/*6, *4/*7, *4/*6, and *6/*6. CONCLUSION: Genotype could be used to screen variations of CYP2D6 expression. PMID- 10678118 TI - Presynaptic release of ATP from superior cervical ganglion of rats modulated by various receptors. AB - AIM: To determine whether adenosine 5'-triphosphate (ATP) is released from the superior cervical ganglion (SCG) of rats and whether the release is regulated by presynaptic mechanism. METHODS: Using the luciferin-luciferase technique. RESULTS: Electric stimulation evoked the release of ATP from the rat SCG. Adenosine (100 mumol.L-1), P1(A1) purinoceptor agonist N6-cyclopentyladenosine (0.1 mumol.L-1), the muscarinic agonist oxotremorine (1 mumol.L-1), and 5 hydroxytryptamine (100 mumol.L-1) decreased the evoked release of ATP from the rat SCG. On the contrary, P1(A1) purinoceptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (10 nmol.L-1), P2 purinoceptor antagonist pyridoxal-5-phosphate 6-azophenyl-2',4-disulphonic acid (10 mumol.L-1), muscarinic antagonist atropine (1 mumol.L-1), alpha 2 adrenoceptor antagonist yohimbine (3 mumol.L-1), D2 dopamine receptor antagonist sulpiride (20 mumol.L-1), and histamine (100 mumol.L 1) increased the evoked release of ATP from the rat SCG. CONCLUSION: ATP is released from the rat SCG and the release of ATP can be presynaptically modulated by P1(A1), P2, muscarinic, alpha adrenergic, D2, 5-HT, and H1 receptor agonists and antagonists. PMID- 10678119 TI - Influence of agmatine in adaptation of cAMP signal transduction system of opiate receptors. AB - AIM: To observe attenuative effects of agmatine on opiate desensitization and substance dependence. METHODS: Guanosine 5'-O-(3-[35S] thiotriphosphate) ([35S]GTTP) binding and cellular cyclic AMP (cAMP) level were determined by radioligand binding assay and radioimmunoassay in NG108-15 cells, respectively. RESULTS: Agmatine increased stimulative action of opioids on [35S]GTTP binding by about 35% and inhibitory effects of opioids on cellular cAMP concentration by about 114.3% in NG108-15 cells pretreated with opioids. On the other hand, it also inhibited cAMP over-shooting by 214.9% of morphine substance dependent cells precipitated by naloxone compared with that of control. These effects of agmatine were antagonized by idazoxan in a concentration-dependent manner. CONCLUSION: Agmatine reversed the formative process of adaptation in cAMP signal transduction cascade. PMID- 10678120 TI - Effects of fenfluramine combined with electroacupuncture on monoamine release in periaqueductal gray of rat brain. AB - AIM: To study the changes of monoamines in ventrolatoral periaqueductal gray of rat brain before and after electroacupuncture (EA) analgesia (EAA) was enhanced by fenfluramine (Fen), a 5-hydroxytryptamine (5-HT) releaser. METHODS: Monoamines were collected by in vivo microdialysis and measured by HPLC connected with electrochemical detector. RESULTS: The level of norepinephrine (Nor) after EA was decreased (P < 0.05 vs NS group). The contents of 5-HT, 5-hydroxyindol acetic acid (5-HIAA), dopamine (DA), and homovanillic acid (HVA) in periaqueductal gray dialysate were increased (P < 0.05 vs NS group). When Fen was combined with EA, the level of 5-HT and 5-HIAA were further increased (P < 0.05 vs NS + EA group). There was no obvious change of Nor, DA, and HVA. CONCLUSION: Fen potentiating EAA may be related to further activation of serotoninergic system. PMID- 10678121 TI - Huperzine-A capsules enhance memory and learning performance in 34 pairs of matched adolescent students. AB - AIM: To study the efficacy of huperzine-A capsules (Hup) on memory and learning performance of adolescent students. METHODS: Using double-blind and matched pair method, 34 pairs of junior middle school students complaining of memory inadequacy were divided into two groups by normal psychological health inventory (PHI), similar memory quotient (MQ), same sex and class. The Hup group was administrated orally 2 capsules of Hup (each contains Hup 50 micrograms) b.i.d., and the placebo group was given 2 capsules of placebo (starch and lactose inside) b.i.d. for 4 wk. RESULTS: At the end of trial, the Hup group's MQ (115 +/- 6) was more than that of the placebo group (104 +/- 9, P < 0.01), and the scores of Chinese language lesson in the Hup group were elevated markedly too. CONCLUSION: The Hup capsules enhance the memory and learning performance of adolescent students. PMID- 10678122 TI - Effect of moxonidine on carotid sinus baroreflex in anesthetized rats. AB - AIM: To study the effect of moxonidine (Mox) on carotid sinus baroreflex. METHODS: By perfusing the carotid sinus in anesthetized rats, the functional parameters of baroreflex were measured. The femoral artery was perfused with constant flow and the change of perfusing pressure was recorded to determine the effect of Mox on vascular tone. RESULTS: Mox 32 and 100 mumol.L-1 shifted the functional curve of carotid sinus baroreflex to the right and upward, with the reduction in peak slope and in reflex decrease of mean arterial pressure, suggesting that Mox produced an inhibitory effect on baroreflex. The effect of Mox 100 mumol.L-1 on baroreflex was completely blocked by efaroxan 100 mumol.L-1. Mox increased vascular resistance. CONCLUSION: Mox inhibits carotid baroreflex via its constrictive action on sinus wall. PMID- 10678123 TI - Effects of allitridi on intracellular Ca2+ concentration in isolated rat brain cells. AB - AIM: To study actions of allitridi extracted from garlic on intracellular calcium in isolated rat brain cells. METHODS: Brain cells were isolated from newborn rat brain with Fura 2-AM measurements of intracellular Ca2+ concentration ([Ca2+]i). RESULTS: Allitridi 1-100 mumol.L-1 concentration-dependently blocked increases of [Ca2+]i caused by potassium chloride and sodium glutamate (Glu) with IC50 of 59.7 and 69.9 mumol.L-1 respectively. Allitridi 100 mumol.L-1 blocked norepinephrine (Nor)-induced [Ca2+]i elevation. CONCLUSION: Allitridi is an effective agent for blocking the [Ca2+]i increase caused by potassium chloride, Nor and Glu. PMID- 10678124 TI - Target selectivity of MAPK phosphorothioate antisense ODN on p42/p44, p38 MAPK, and JNK protein expression and its inhibitory effect on VSMC DNA synthesis. AB - AIM: To analyze the target selective and sequence-specific inhibitory effect of mitogen-activated protein kinase (MAPK) phosphorothioate antisense oligodeoxynucleotides (ODN) on p42/p44, p38 MAPK, c-jun NH2-terminal protein kinases (JNK) protein expression, and DNA synthesis in vascular smooth muscle cell (VSMC). METHODS: Using a phosphorothioate-protected 17-mer antisense MAPK ODN directed against the initiation of translation sites of the p42/p44 MAPK isoforms by liposomal transfection to deplete cultured rat, rabbit, and fetal calf VSMC MAP kinases. The 17-mer sense and random sequence MAPK ODN were used as controls. After liposomal transfection, cells were exposed to 20% serum for 24 h, and then harvested in lysis buffer. P42/p44, p38 MAPK, and p46/p58 JNK protein expression were measured by Western blot. DNA synthesis was measured by [3H]thymidine incorporation. RESULTS: Treatment with MAPK antisense ODN (0.1-0.8 mumol.L-1) for 48 h reduced phosphored p42/p44 MAPK protein expression but without effect on p38 MAPK and JNK expression, and inhibited cultured rat, rabbit, and fetal calf VSMC [3H]thymidine incorporation stimulated by 20% serum in a concentration-dependent manner. CONCLUSION: The MAPK antisense ODN target selectively and sequence-specifically reduces the p42/p44 MAPK protein expression and concentration-dependently inhibits proliferation of rat, rabbit and fetal calf VSMC. PMID- 10678125 TI - Effect of advanced glycosylation end products on diacylglycerol signaling pathway in cultured rat aortic smooth muscle cells. AB - AIM: To investigate whether diacylglycerol (Dia) signaling pathway is stimulated by advanced glycosylation end products (AGEP) and to determine effect of vitamin E and aminoguanidine on Dia level induced by AGEP in cultured rat aortic vascular smooth muscle cells (VSMC). METHODS: The effect of AGEP on Dia level in cultured VSMC was measured by radio-enzymatic assay. Quantitative measurements of [32P]phosphatidic acid were performed by thin-layer chromatography and autoradiography. RESULTS: The Dia level in VSMC incubated with AGEP-BSA was markedly increased in a concentration-dependent, biphasic manner. The early phase was rapid and transient, peaking at 15 s. The late phase reached the maximal level at 10 min and decayed slowly. The Dia levels in VSMC exposed to different concentrations of AGEP-BSA and AGEP-BSA glycosylated for various periods were greatly increased compared with control group. Vitamin E 50, 100 nmol.L-1 prevented the AGEP-BSA-stimulated elevation of Dia level in VSMC, from (940 +/- 43) pmol.L-1 to (599 +/- 38), (290 +/- 21) pmol.L-1, respectively. In aminoguanidine-treated AGEP-BSA groups, Dia level in the cells incubated with the same concentration of AGEP-BSA (100, 200 mg.L-1) were decreased to (260 +/- 8) and (289 +/- 10) pmol.L-1, respectively. Early glycosylated low-density lipoproteins (LDL) did not affect Dia level. CONCLUSION: AGEP causes a robust stimulation of Dia signaling pathway in VSMC. Vitamin E and aminoguanidine attenuate the production of Dia. PMID- 10678126 TI - Inhibitory effects of sodium quercetin monosulfate on pig platelet aggregation induced by thrombin. AB - AIM: To study the inhibitory effects of sodium quercetin monosulfate (SQMS) on pig platelet aggregation induced by thrombin. METHODS: Platelet aggregation was analyzed by turbidimetry. Cytosolic free calcium concentration ([Ca2+]i) was determined by Fura-2 fluorescence. Activity of protein kinase C (PKC) was assayed by incubating PKC with histone III S and [gamma-32 P] ATP. The cytoskeletal proteins were precipitated by Triton X-100 and separated by SDS-PAGE. RESULTS: SQMS inhibited the platelet aggregation induced by thrombin 500 U.L-1 with IC50 132 (50-347) mumol.L-1. SQMS inhibited Ca2+ influx in blood platelets induced by thrombin 500 U.L-1 in the presence of extracellular Ca2+ 1 mmol.L-1 with IC50 20 (9-46) mumol.L-1; SQMS inhibited the internal Ca2+ release in the absence of extracellular Ca2+. SQMS also decreased [Ca2+]i level in quiescent blood platelets. SQMS (10-160 mumol.L-1) inhibited the activity of cytosolic PKC from blood platelets in a concentration-dependent manner, but had no effect on membrane PKC. SQMS (20-80 mumol.L-1) inhibited the actin polymerization induced by thrombin 500 U.L-1 inblood platelets in a concentration-dependent manner. CONCLUSION: SQMS inhibited pig platelet aggregation induced by thrombin and its mechanism might be due to its inhibitions of Ca2+ influx, internal Ca2- release, PKC activity, and actin polymerization. PMID- 10678127 TI - Effects of MK-447 on platelet shape change, aggregation, and ATP release by collagen, ADP, and stable analogue of thromboxane A2 in rabbit platelets. AB - AIM: To investigate the effects of MK-447 on platelet shape change, aggregation, and ATP release by collagen (Col), ADP, and stable analogue of thromboxane A2 (STA2) in rabbits. METHODS: Platelet shape change and aggregation were quantified in light transmission by turbidimetric method and release reaction was assessed by the amount of ATP in platelet-rich plasma (PRP). RESULTS: (1) MK-447 100-700 mumol.L-1 caused only the shape change, which was not inhibited by indometacin 3 mumol.L-1. Platelet shape changes by Col, ADP, and STA2 were reduced (P < 0.01) after the addition of MK-447. The lag phase was prolonged (P < 0.01) in Col and shortened (P < 0.01) in ADP. (2) MK-447 reduced the aggregation by Col 5 mg.L-1 (P < 0.01), and enhanced that by ADP 0.3-10 mumol.L-1 and STA2 0.1-3 mumol.L-1 (P < 0.01). (3) The release reaction by STA2 1-3 mumol.L-1 was also increased (P < 0.01). The effects of MK-447 on STA2 were not inhibited by S-145. CONCLUSION: MK 447 induced the platelet shape change, and showed the dual effects, inhibition or enhancement, on the actions by different aggregating agents. PMID- 10678128 TI - Inhibitory effects of estradiol on inward rectifier and delayed rectifier K+ currents in guinea pig ventricular myocytes. AB - AIM: To study the effects of estradiol (Est) on inward rectifier K+ (IK1) and delayed rectifier K+ (IK) channels in isolated guinea pig ventricular myocytes. METHODS: Using whole cell patch-clamp recording techniques. RESULTS: Est 10 mumol.L-1 and 100 mumol.L-1 decreased the action potential duration, APD50, from (474 +/- 71) ms to (330 +/- 75) ms and (229 +/- 67) ms (n = 7 cells of 7 guinea pigs, P < 0.05), respectively. Est 100 mumol.L-1 also decreased APD90 from (587 +/- 60) ms to (418 +/- 79) ms (n = 7, P < 0.05). Est inhibited IK tail current (IK.tail) concentration-dependently. IK.tail was depressed 53% (n = 5, P < 0.05) at 10 mumol.L-1 and 80% (n = 5, P < 0.01) at 100 mumol.L-1 compared with control. Est > or = 10 mumol.L-1 blocked IK1. The maximal inhibition of inward current of IK1 occurred at -100 mV test potential was 49% (n = 5, P < 0.01) and outward current of IK1 at -40 mV was 72% (n = 5, P < 0.01). The reverse potential shifted negatively, from -70 to -76 mV. CONCLUSION: Est possessed blocking effects on both IK1 and IK channels in guinea pig ventricular myocytes. PMID- 10678130 TI - Hypotensive effect of tenuifolic saponin and its mechanism. AB - AIM: To study the effect of tenuifolic saponin (TS) on arterial pressure. METHODS: Mean arterial pressure (MAP) was recorded from left carotid artery in rat which was anesthetized with urethane and then injected i.v. gtt with a transfusion of NaCl 0.15 mol.L-1. Systolic blood pressure (SBP) of conscious rat and renovascular hypertensive rat (RVHR) was measured by tail cuff method. RESULTS: TS 2, 4, 8 mg.kg-1 i.v., 20 and 40 mg.kg-1 i.g. reduced the MAP by 31%, 37%, 50%, 21%, and 31%, respectively. Bilateral vagotomy plus atropine (Atr) i.v., or pretreatment with diphenhydramine hydrochloride (Dip) failed to influence TS effect. Lack of effect of TS on carotid-occlusion-induced- or epinephrine (Epi)-induced-hypertensive response was found. SBP in conscious rat and RVHR was suppressed, highest by 38.0% and 26.8% at 60 and 90 min, maintaining at least 2 and 3 h, respectively, after i.g. TS 40 mg.kg-1. CONCLUSION: TS reduced the arterial pressure, not related to vagus excitation, ganglionic blockade, and peripheral alpha-adrenergic-, M-cholinergic-, and H1-receptors. PMID- 10678129 TI - Protective effects of Ginkgo biloba extract on cultured rat cardiomyocytes damaged by H2O2. AB - AIM: To investigate the influence of Ginkgo biloba extract (GbE) on cardiomyocytes damaged by H2O2. METHODS: Cultured rat cardiomyocytes were divided into 3 groups randomly: control group; H2O2 (2.5 mmol.L-1) group; H2O2 2.5 mmol.L 1 + GbE 150 mg.L-1 group. The cardiomyocytes were cultured in MEM (Eagle's) at 37 degrees C in the presence of 5% CO2 for 4 h. Lactate dehydrogenase (LDH) was assayed by colorimetric method. Lipid peroxidation was determined by measuring thiobarbituric acid-reactive substances. Ultrastructure was viewed under transmission electron microscope. RESULTS: Compared with the control group, LDH leakage and malondialdehyde (MDA) content increased in H2O2 group, LDH increased from (2166 +/- 247) U.L-1 to (5180 +/- 648) U.L-1, MDA increased from (3.5 +/- 0.2) nmol/10(6) cells to (7.2 +/- 0.4) nmol/10(6) cells (P < 0.01). The ultrastructure was damaged seriously. GbE inhibited the increase of LDH leakage and MDA content induced by H2O2. In this group, LDH decreased from (5180 +/- 648) U.L-1 to (3496 +/- 386) U.L-1, MDA decreased from (7.2 +/- 0.4) nmol/10(6) cells to (4.8 +/- 0.9) nmol/10(6) cells (P < 0.01). Ultrastructure of cells was also protected by GbE. CONCLUSION: GbE protected the cardiomyocyte against H2O2 injury, the protective action was attributed to its antiperoxidative effect. PMID- 10678131 TI - Glutathione antagonized cyclophosphamide- and acrolein-induced cytotoxicity of PC3 cells and immunosuppressive actions in mice. AB - AIM: To study the antagonistic effect of glutathione (GSH) on toxicity of PC3 cell induced by cyclophosphamide (Cyc) and acrolein (Acr) and on immunosuppressive actions caused by Cyc. METHODS: Splenocyte, PC3 cell proliferation and cell protein content were measured by tetrazolium (MTT) assay and Coomassie brilliant blue assay. Serum anti-SRBC hemolysin, agglutinin, and splenocyte proliferation were measured in normal and S-180-bearing mice. Tumors were weighed. RESULTS: Pretreatment with GSH 2 mmol.L-1 reduced splenocyte proliferation inhibition from 18.64%, 49.72% to 6.78%, 18.36% (induced by Cyc 1, and 5 mmol.L-1), and PC3 cell proliferation inhibition from 27.7%, 45.3%, and 74.6% to 14.6%, 18.8%, and 49.1% (induced by Cyc 1, 3, and 5 mmol.L-1), and from 62.6%, 85.4%, and 90.6% to 41.9%, 57.7%, and 86.4% (induced by Acr 10, 25, and 50 mumol.L-1), respectively. In normal mice, s.c. GSH 75 or 150 mg.kg-1 b.i.d. x 5 d after i.p. Cyc 40 mg.kg-1, the hemolysin and the splenocyte proliferation were higher than those in normal mice i.p. Cyc 40 mg.kg-1 alone. Hemolysin, serum agglutinin, and splenocyte proliferation in S-180-bearing mice given s.c. GSH 150 mg.kg-1 b.i.d. x 10 d after i.p. Cyc 40 mg.kg-1 were also markedly higher than those in S-180-bearing mice given i.p. Cyc alone. But, according to tumor weight, GSH did not interfere the antitumor activity of Cyc in S-180-bearing mice. CONCLUSION: GSH exhibited protective effects against Cyc and Acr, but had no effect on the antitumor action of Cyc. PMID- 10678132 TI - R-dl-verapamil downmodulates multidrug resistance of KBv200 cells to vincristine and doxorubicin. AB - AIM: To study the attenuation of multidrug resistance (MDR) by R-dl-verapamil (R Ver) and the acute animal toxicity of R-Ver, and to compare these results of R Ver with the results of dl-verapamil (Ver). METHODS: Cytotoxicity was determined by tetrazolium (MTT) assay. Cellular accumulation of doxorubicin (Dox) was measured by fluorescence spectrophotometry. Acute animal toxicity was tested by i.p. drug administration in BALB/c mice. RESULTS: R-Ver attenuated MDR of KBv200 cells to vincristine (VCR) and Dox. This attenuation ability was dose-related, and was also dependent on drug exposure time. R-Ver 1.25 mumol.L-1 increased the sensitivity of KBv200 cells to VCR (P < 0.01) with a 24-h period of drug exposure. R-Ver downmodulated MDR and increased cellular Dox accumulation of KBv200 cells as effectively as Ver, but possessed lower acute toxicity in BALB/c mice. While LD50 of Ver was 60 (49-73) mg.kg-1, LD50 of R-Ver was 166 (137-202) mg.kg-1. CONCLUSION: R-Ver downmodulated the MDR to VCR and Dox at 1.25 mumol.L 1, and this effect on VCR can be realized with drug exposure duration of 24 h. PMID- 10678133 TI - Bronchodilating effects of bambuterol on bronchoconstriction in guinea pigs. AB - AIM: To study the effects of bambuterol (Bam) on bronchoconstriction in guinea pigs. METHODS: Bronchospasm induced by histamine aerosol, lung resistance (RL) and dynamic lung compliance (Cdyn) changes induced by ovalbumin aerosol in vivo, isolated resting lung parenchyma strips, and carbamylcholine-induced tracheal constriction in vitro in guinea pig were investigated. RESULTS: Bam dose dependently prolonged the time to histamine-induced collapse, ED50 values (95% confidence limits) of Bam intragastric gavage (i.g.) after 1 h, 4 h, and 24 h were 0.74 (0.60-0.91), 0.75 (0.61-0.91) and 1.00 (0.77-1.30) mg.kg-1, respectively. Bam 2 or 10 mg.kg-1 i.g. 2 h before ovalbumin aerosol partly or almost completely inhibited bronchial challenge of ovalbumin-induced change of RL and Cdyn. Bam 0.1-1.0 mumol.L-1 gave a weak relaxation on isolated tracheal strips induced by carbamylcholine and failed to relax the isolated resting lung parenchyma strips in guinea pig. CONCLUSION: Bam showed a long-acting bronchodilation by its slow metabolism in vivo. PMID- 10678134 TI - Pharmacokinetics of 2-hydroxyflutamide, a major metabolite of flutamide, in normal and CCl4-poisoned rats. AB - AIM: To study the pharmacokinetics of 2-hydroxyflutamide (HF), a major active metabolite of flutamide (Flu), in normal and CCl4-poisoned rats. METHODS: Normal and CCl4-poisoned rats were given i.g. HF 25 mg.kg-1. HF concentrations of plasma were determined by HPLC with YWG C 18 column, Flu was used as an internal standard. The mobile phase was composed of methanol: water = 3:2 (vol), and absorbance was measured at lambda 295 nm. RESULTS: HF elimination was inhibited in CCl4-poisoned rats compared with normal rats. K decreased from (0.11 +/- 0.05) to (0.05 +/- 0.01) h-1 (P < 0.01), T1/2 was prolonged from (6.8 +/- 1.9) to (14 +/- 4) h (P < 0.01), Cl decreased from (0.18 +/- 0.06) to (0.12 +/- 0.02) L.kg 1.h-1 (P < 0.05), AUC increased from (149 +/- 47) to (226 +/- 54) mg.L-1.h (P < 0.05). CONCLUSION: This HPLC assay was sensitive and precise, and the elimination of HF was inhibited due to CCl4 poisoning. PMID- 10678135 TI - [Effects of acetylcholine on inositol 1,4,5-triphosphate formation in cervix uteri myometrial cells of human]. AB - AIM: To detect inositol 1,4,5-triphosphate (IP3) formation of pregnant and nonpregnant human myometrial cells induced by acetylcholine (ACh). METHODS: [3H] IP3 competitive protein binding assay. RESULTS: Basal levels of IP3 in pregnant and nonpregnant human myometrial cells were (82 +/- 9) and (96 +/- 10) nmol.g-1 (protein), respectively (n = 6). Incubated with ACh 50 mumol.L-1 for 5 min, IP3 reached the peak levels (109 +/- 11) and (122 +/- 15) nmol.g-1 (protein), respectively (n = 6), but difference of the increments of IP3 in pregnant and nonpregnant women was not significant. The increased IP3 induced by ACh was inhibited by atropine (Atr) 1 mumol.L-1. CONCLUSION: The basal IP3 level in pregnant cervix myometrial cells was higher than that in nonpregnant women. ACh increased the IP3 formation. PMID- 10678136 TI - Effects of ramipril on cardiac gene transcription levels of angiotensin II receptors after myocardial infarction. AB - AIM: To study the early changes of cardiac angiotensin (Ang) II receptor gene transcription after myocardial infarction (MI) in rats chronically treated with the angiotensin-converting enzyme (ACE) inhibitor ramipril. METHODS: MI was induced by left anterior descending coronary artery ligation in rats and sham operated rats were used as control. Rats were treated daily with ramipril (1 mg.kg-1) or water, initiated 1 wk before surgery. Quantitative RT-PCR was applied to determine the Ang II receptors AT1, AT2 receptor gene mRNA levels in the non infarcted myocardium. RESULTS: AT1 and AT2 mRNA levels increased time point dependently in the cardiac septum after MI reaching a peak on d 1. There was no significant difference of the myocardial AT1 and AT2 receptor mRNA levels between the ramipril-treated and water-treated rats after MI. CONCLUSION: The AT1 and AT2 receptor gene transcription in the non-infarcted myocardium was associated with the process of cardiac remodeling after MI but not affected by ACE inhibition. PMID- 10678137 TI - Huperzine-A in capsules and tablets for treating patients with Alzheimer disease. AB - AIM: To compare the efficacy and safety between huperzine-A (Hup) in capsules and tablets for treating patients with Alzheimer disease (AD). METHODS: Using multicenter, prospective, double-blind, double-mimic, parallel, positive controlled and randomized methods, 60 patients meeting with the NINCDS-ARDRA criteria of AD were divided into 2 equal groups. Patients in the capsule group received 4 capsules of Hup (each contains 50 micrograms) and 4 tablets of placebo (lactose and starch inside); while the tablet group received 4 tablets of Hup (each contains 50 micrograms) and 4 capsules of placebo, p.o., twice a day for 60 d. All the patients were evaluated with a lot of related ranting scales, and physiological and laboratory examination. RESULTS: There were significant differences (P < 0.01) on all the psychological evaluations between 'before' and 'after' the 60-d trial of 2 groups, but there was no significant difference between 2 groups by group t test (P > 0.05). The changes of oxygen free radicals in 2 groups showed marked improvement. No severe side effect besides moderate to mild nausea was found in both groups. CONCLUSION: There is equal efficacy and safety between Hup in capsule and tablet for treating patients with AD, and Hup can reduce the pathological changes of the oxygen free radicals in the plasma and erythrocytes of patients with AD. PMID- 10678138 TI - Leucine-2-alanine enkephalin induced delta opioid receptors internalization expressed stably in CHO cells. AB - AIM: To characterize the internalization of delta opioid receptors (DOR) stably expressed in Chinese hamster ovary (CHO) cells and the role of the C-terminal in this process. METHODS: Receptor membrane anchoring was shown by immunofluorescence microscopy. Receptor internalization was assessed by measuring the radioligand binding resistant to the acid-buffer wash. RESULTS: Originally, all the wild-type (CHO-W) and C-truncated (CHO-T) DOR expressed were localized to the membrane. Agonist [3H] leucine-2-alanine enkephalin (LAE) but not the antagonist [3H]diprenorphine (Dip) induced rapid receptor internalization. The internalization of C-truncated DOR in CHO-T was similar to that of the wild-type in maximal level, but climbed up more slowly. DOR internalization was extracellular osmolarity- and temperature-sensitive. Pertussis toxin and universal protein kinase inhibitor staurosporine had no effect on it. CONCLUSION: DOR internalization is an agonist and clathrin-coated pits dependent, but post receptor cellular signal transduction independent process; moreover, the C terminal of DOR, not engaged in membrane anchoring, affects the initialization of DOR internalization. PMID- 10678139 TI - Recurrent events in meta-analysis of multiple clinical trials. AB - AIM: To study the efficacy and safety of drug or therapy with recurrent events in meta-analysis of multiple clinical trials. METHODS: A nonparametric approach is proposed to estimate the rates of recurrent events for meta-analysis of trials. The method was used in meta-analysis of angiotensin-converting enzyme (ACE) inhibitor clinical trials to analyze the relative rates and the excess rates between ACE inhibitor and placebo treatment groups for endpoints of hospitalizations for CHF, hospitalizations for CHF or cardiac death, and hospitalizations for CHF or any death, respectively. RESULTS: The estimates of those three endpoints were 69%, 74%, and 76% (P < 0.01). Compared with placebo, ACE inhibitor reduced 30 cases of hospitalizations for CHF per 1000 person-years, or 40 cardiac deaths or hospitalizations for CHF per 1000 person-years (P < 0.01). CONCLUSION: The method was a simple and efficient approach to conduct meta analysis of clinical trials with recurrent events. PMID- 10678140 TI - Effects of low-pH treatment on cAMP second messenger system regulated by different opioid agonists. AB - AIM: To study the mechanism of opioid agonists in regulation of cAMP second messenger system. METHODS: Low-pH treatment was used to deplete the stimulatory G protein (Gs) function. The effects of some opiates on adenylate cyclase were compared between control and low-pH treatment membranes. RESULTS: In contrast to dehydroetorphine (DHE), etorphine (Eto), morphine (Mor) and methadone (Met) substantially increased the inhibitory effects on adenylate cyclase in membranes prepared from naive and chronic Mor- or Met-treated NG108-15 cells by low-pH treatment. In contrast to Mor, DHE and Eto did not result in significant decrease in the inhibitory effects on adenylate cyclase in membranes from the cells treated chronically with DHE or Eto. Marked rebound of adenylate cyclase was also not observed in membranes from chronic DHE or Eto-treated cells when precipitated with naloxone. Low-pH treatment eliminated naloxone-induced rebound of adenylate cyclase in chronic Mor-treated cells. CONCLUSION: The difference in opiate induced functional adaptive alteration of Gs is at least one biochemical mechanism of developing opiate tolerance and dependence. PMID- 10678141 TI - 5-HT1P receptor-mediated slow depolarization in neurons of guinea pig inferior mesenteric ganglion. AB - AIM: To study the effects of several 5-hydroxytryptamine (5-HT) receptor subtype antagonists on 5-HT-induced depolarization and the effects of 5-HT1P receptor agonist on the membrane potential in the neurons of guinea pig inferior mesenteric ganglion (IMG). METHODS: Intracellular recordings were made from neurons of the isolated guinea pig IMG. RESULTS: Cyproheptadine (5-HT1/2 antagonist 10 mumol.L-1, n = 7) and BRL 24924 (5-HT1P antagonist 10 mumol.L-1, n = 19) reversibly suppressed 5-HT slow response; pressure ejection of MCPP (5-HT1P agonist 10 mmol.L-1) induced a slow depolarization in most of 5-HT sensitive neurons (10/14). CONCLUSION: 5-HT-induced slow depolarization is mediated by 5 HT1P receptor. PMID- 10678142 TI - dl-3-n-butylphthalide improves regional cerebral blood flow after experimental subarachnoid hemorrhage in rats. AB - AIM: To investigate the effect of dl-3-n-butylphthalide (dl-NBP) on experimental subarachnoid hemorrhage (SAH) in rats. METHODS: SAH was induced by injection of autologous artery blood 0.35 mL into lateral ventricle. Regional cerebral blood flow (rCBF) in caudate nucleus was determined by hydrogen clearance method. RESULTS: A rapid and marked decrease in rCBF in caudate nucleus was observed 15 min after SAH and the rCBF remained at low level (about 50% pre-SAH value) within 180 min. dl-NBP (50, 100 mg.kg-1, i.g.) increased rCBF 30-180 min after the onset of SAH without significant effect on mean artery blood pressure. dl-NBP 100 mg.kg 1 increased rCBF in caudate nucleus by 26% at 15 min and by 36% at 180 min respectively after SAH. d-NBP but not l-NBP (10 mg.kg-1, i.p.) increased rCBF. CONCLUSION: dl-NBP improves rCBF in caudate nucleus of rats subjected to SAH. PMID- 10678143 TI - Dauricine suppressed CsCl-induced early afterdepolarizations and triggered arrhythmias in rabbit heart in vivo. AB - AIM: To study the effect of dauricine on CsCl-induced early afterdepolarizations (EAD) and ventricular arrhythmias in rabbits. METHODS: Monophasic action potentials (MAP) of the left ventricle of the rabbit heart in situ were recorded with MAP recording technique. CsCl 1-2 mmol.kg-1 i.v. was used to induce EAD and ventricular arrhythmias. RESULTS: CsCl resulted in decrease of MAP amplitude (MAPA, P < 0.05) and prolongation of MAP duration at 90% repolarization (MAPD90, P < 0.01), QRS, and R-R duration (P < 0.05) compared with those before CsCl in the dauricine and control group. CsCl injection induced EAD that appeared within about 30 s and disappeared 5-15 min thereafter. EAD always preceded ventricular arrhythmias including ventricular premature beats and paroxysmal ventricular tachycardia. The EAD amplitude (EADA) in the dauricine group (26% +/- 9% of MAPA) was smaller than that in the control group (52% +/- 5% of MAPA, P < 0.05) and the incidence of arrhythmias in dauricine group (28%) was lower than that in control group (80%, P < 0.05). CONCLUSION: Dauricine exerted an antagonistic effect on EAD and suppressed triggered ventricular arrhythmias by decreasing EADA. PMID- 10678144 TI - Anti-arrhythmic effects of sophoridine and oxysophoridine. AB - AIM: To compare the effects of oxysophoridine (Oxy) and sophoridine (Sop) on experimental arrhythmias and myocardial physiologic properties. METHODS: Arrhythmias were induced by drugs and myocardial ischemia. Physiologic properties were determined on isolated heart atria. RESULTS: Oxy 500 mg.kg-1 (1/6 LD50) decreased the incidence of ventricular arrhythmias induced by aconitine (P < 0.01), increased the threshold dose of ouabain-induced ventricular premature (VP, P < 0.05), ventricular tachycardia (VT, P < 0.05), ventricular fibrillation (VF, P < 0.01), and cardiac arrest, (P < 0.01). After i.v. Oxy 500 mg.kg-1 into the rats with ligation of left anterior descending coronary artery, the total numbers of ectopic beats were decreased (P < 0.05), the incidence of VF was lowered, and the duration of VT was shortened (P < 0.01). Oxy 250 mg.kg-1 (1/13 LD50) i.v. shortened the duration of arrhythmias induced by BaCl2 (P < 0.01) and delayed the onset of arrhythmias induced by chloroform-epinephrine (P < 0.05). Oxy produced dose-dependent positive inotropic effects in the isolated left atrial of guinea pigs, increased the concentration of epinephrine to elicit automaticity in left atria, decreased slightly the excitability, and prolonged the functional refractory period. Sop produced the similar effects on arrhythmias as Oxy. CONCLUSION: Oxy produced the similar anti-arrhythmic effects as Sop did at the equivalent effective dose. PMID- 10678145 TI - High glucose enhances mitogenic response to endothelin-1 in rabbit vascular smooth muscle cells. AB - AIM: To examine the effects of high glucose on the mitogenic response of rabbit aortic vascular smooth muscle cells (VSMC) to endothelin-1 (ET-1). METHODS: VSMC were cultured in normal glucose (5.5 mmol.L-1), high glucose (25 mmol.L-1) or high osmolality (glucose 5.5 mmol.L-1, plus mannitol 19.5 mmol.L-1). DNA synthesis was measured by [3H]thymidine incorporation. The expression of phospho p44/42 MAPK was determined by Western blot. RESULTS: At a concentration range from 10(-12) to 10(-8) mol.L-1, ET-1 stimulated [3H]thymidine incorporation and phospho-p44/42 MAPK expression in VSMC in a concentration-dependent manner. From 10(-11) to 10(-8) mol.L-1, the mitogenic effect of ET-1 was higher in VSMC cultured in high glucose at equivalent concentration than cells cultured in normal glucose or high osmolality (P < 0.05 or P < 0.01), but no marked difference was observed in the growth response between cells cultured under the latter two conditions. Similarly, ET-1 increased expression of phospho-p44/42 MAPK by 60%-65% in VSMC cultured in high glucose, compared with cells in normal glucose or high osmolality. CONCLUSION: VSMC cultured in high glucose exhibited increased mitogenic response to ET-1, which seemed to be related to the enhanced expression of phospho-p44/42 MAPK. PMID- 10678146 TI - Inhibition of myocardial inward rectifier potassium current by propylbutyldopamine. AB - AIM: To study the effects of propylbutyldopamine (PBDA) on the inward rectifier potassium current (Ik1). METHODS: The quasi-steady state current-voltage relationship from the isolated guinea pig ventricular cells were measured using whole-cell patch-clamp techniques with a slow ramp depolarization (8 mV.s-1). RESULTS: PBDA 5, 50, and 100 mumol.L-1 concentration-dependently reduced the inward rectifier potassium current. PBDA blocked Ik1 in guinea pig ventricular cells. The effect of PBDA was not blocked by the selective dopamine D2-receptor blocker, domperidone. CONCLUSION: PBDA inhibited Ik1 directly, independent of the dopamine D2-receptor. PMID- 10678147 TI - Prophylactic effects of tetramethyl pyrazine on mice with endotoxemia and its relationship with platelet-activating factor. AB - AIM: To study the prophylactic effects of tetramethyl pyrazine (TMP) on mice with endotoxemia and its relationship with platelet-activating factor (PAF). METHODS: LD80 of lipopolysaccharides (LPS, 15 mg.kg-1) was injected into mice (i.v.) pretreated with TMP (i.p.). The survival rate and the level of serum PAF were observed. The PAF induced by LPS in vivo and in vitro, and the activities of PLA2 and acetyl-CoA: lyso-PAF acetyltransferase were determined. RESULTS: TMP (10, 30, 90 mg.kg-1) obviously lowered the mortality of mice and also dose-dependently decreased the level of serum PAF [(25.5 +/- 1.7), (13.4 +/- 3.2), (9.6 +/- 2.1) micrograms.L-1, vs control (29.3 +/- 2.1) micrograms.L-1, P < 0.01]. TMP (0.05, 0.5, 5, 50 mumol.L-1) dose-dependently decreased the release of PAF [(12.7 +/- 1.6), (8.9 +/- 1.2), (6.9 +/- 0.8), (5.5 +/- 1.0) micrograms.L-1 vs control (11.9 +/- 1.8) micrograms.L-1, P < 0.01] from PMO cultured with LPS (5 mg.L-1), reduced the PLA2 activity [(149.9 +/- 2.8), (117.5 +/- 2.0), (89.6 +/- 2.0), (75.0 +/- 2.8) U vs control (170.8 +/- 3.9) U, P < 0.01] and acetyl-CoA: lyso-PAF acetyltransferase activity [PAF (9.5 +/- 0.7), (5.2 +/- 0.7), (2.9 +/- 0.3), (2.5 +/- 0.3) micrograms.g-1 (protein).min-1 vs control (11.0 +/- 0.7) micrograms.g-1 (protein).min-1, P < 0.01] of PMO lysate. CONCLUSION: TMP protected the mice with endotoxemia from the death by decreasing the biosynthesis of PAF through the inhibition of the activities of PLA2 and acetyl-CoA: lyso-PAF acetyl-transferase. PMID- 10678148 TI - Dauricine inhibits redistribution of platelet membrane glycoprotein IV and release of intracellular alpha-granule thrombospondin induced by thrombin. AB - AIM: To study the possibility of dauricine (Dau) inhibiting redistribution of platelet membrane glycoprotein IV (GPIV) and release of intracellular alpha granule thrombospondin (TSP) on platelet activation. METHODS: Using the flow cytometric assay of washed platelet to record expression of GPIV and release of TSP induced by thrombin. RESULTS: Dau did not affect GPIV and TSP on resting platelet membrane but inhibited redistribution of GPIV to the platelet surface and TSP release on activated platelet. There was a marked positive correlation between changes of GPIV and TSP (r = 0.511, P < 0.01). The inhibitory effect of Dau appeared not to be Ca2+ concentration-dependent. CONCLUSION: Dau inhibited redistribution of GPIV and release of intracellular alpha-granule thrombospondin induced by thrombin. PMID- 10678149 TI - Flutamide suppressed prostate hypertrophy in rats and mice. AB - AIM: To study the suppressive effect of flutamide (Flu) on benign prostate hypertrophy. METHODS: The effect of Flu 10, 25, and 50 mg.kg-1 i.g. on the prostate was tested in orchiectomized rats with s.c. testosterone daily for 30 d and in mice implanted with homologous strain fetal mouse urogenital sinus for 14 d. RESULTS: 1) Flu dose-dependently suppressed the weight and volume of each lobe of the prostate to about 10%-50% of control. Also, the acini and height of epithelial cells atrophied. The effect was more powerful than that of estradiol (Est). 2) The weight and volume of the mouse prostate diminished in Flu-treated groups, but the dose-response relationship was seen only in volume. In this model, Est was better than Flu. CONCLUSION: Flu possesses the suppressive action on benign prostate hypertrophy. PMID- 10678150 TI - Effects of antitumor compounds isolated from Pteris semipinnata L on DNA topoisomerases and cell cycle of HL-60 cells. AB - AIM: To study the effect of the antitumor compounds 5F, 6F, and A from Pteris semipinnata L on the activities of DNA topoisomerases and cell cycle of HL-60 cells, and the synergism of compound 6F in combination with genistein in vitro. METHODS: DNA topoisomerases were isolated from HL-60 cell lines, and supercoiled pBR322 DNA was used as substrate to determine the activities of DNA topoisomerase I and II. Cell cycle was analyzed by flow cytometry (FCM). Cytotoxicity assay was tested by MTT method. RESULTS: Compounds 5F, 6F, and A inhibited the activities of DNA topoisomerase I and II. After exposure of the cells to compound 6F, an increase in cells in the S and G2/M phases and a decrease in cells in the G0/G1 phase of the cell cycle were observed. At low concentrations (57.8 and 115.6 nmol.L-1), compound 6F enhanced the cytotoxicity against HL-60 cell line in combination with genistein, q values were > 1.15. The enhancement times of 57.8 and 115.6 nmol.L-1 of 6F by genistein were 2.60 and 4.65, respectively. CONCLUSION: Compounds 5F, 6F, and A inhibited the activities of DNA topoisomerases of HL-60 cells. Compound 6F increased the number of cells in S and G2/M phases, decreased the population of G0/G1 phase cells, and enhanced the cytotoxicity of genistein, which had synergism with 6F in antitumor action. PMID- 10678151 TI - Suppressive effect of kanglemycin C on T- and B-lymphocyte activation. AB - AIM: To elucidate the suppressive effect of kanglemycin C (Kan) on lymphocyte proliferation and T-lymphocyte subsets. METHODS: Splenocyte proliferation was quantified with [3H]thymidine ([3H]TdR) pulsing method or 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetery. L3T4+ and Lyt2+ T-cell subsets were measured with fluorescence-activated cell sorter (FACS). Splenocyte viability was assessed with trypan blue exclusion. RESULTS: Like ciclosporin (Cic), Kan 8, 40, 80, and 400 nmol.L-1 inhibited the proliferation of 20%-80% incubated mouse splenocytes stimulated by concanavalin A (Con A) 5 mg.L-1, phytohemagglutinin (PHA) 5 mg.L-1, tetradecanoylphorbol acetate (TPA) 10 micrograms.L-1 + ionomycin (IM) 0.5 mg.L-1, and alloantigen (mixed lymphocyte reaction). Kan had no toxicity to the splenocytes at the treated doses. Suppression by Kan was declined with addition time of Kan after culture onset. Furthermore, the suppressive effect of Kan on splenocyte proliferation stimulated by lipopolysaccharides (LPS) 10 mg.L-1 was similar to that on splenocyte proliferation mediated by Con A. Unlike Cic, Kan reversed the ratio of L3T4+/Lyt2+ T-cell subsets. CONCLUSION: Kan had a suppressive action on proliferation of T- and B-lymphocytes and had a selective effect on helper inducer T-lymphocyte (Th) subset from Cic. Suppression by Kan was time-dependent and not associated with toxicity of Kan. PMID- 10678152 TI - Anti-inflammatory effects of total saponins of Panax notoginseng. AB - AIM: To study the anti-inflammatory effects of total saponins of Panax notoginseng (PnS). METHODS: Rat air-pouch acute inflammatory model was established with s.c. carrageenan (Car, 25 mg.kg-1). The protein content in exudate was measured. Micro-acid titration assay and radioimmunoassay (RIA) were applied respectively to investigate effects of PnS on phospholipase A2 (PLA2) activity and dinoprostone (Din) content in exudate. Fura-2 fluorescence technique was used to determine the intracellular free calcium concentration in neutrophils (Neu-[Ca2+]i). RESULTS: At 12 h, PnS 60-240 mg.kg-1 i.p. reduced Neu counts, protein content [(7.7 +/- 1.3) to (4.4 +/- 1.4) g.L-1], and Din content [(1619 +/ 391) to (883 +/- 268) ng.L-1]; inhibited the PLA2 activity in exudate [(248 +/- 42) to (157 +/- 35) kU.L-1] in a dose-dependent manner. PnS 60, 120, and 240 mg.kg-1 lowered the level of Neu-[Ca2+]i with the inhibitory rate of 9.1%, 33.2%, and 39.4%, respectively. CONCLUSION: PnS has an obvious anti-inflammatory effect and its mechanisms are related to the inhibition of the Neu-[Ca2+]i level and PLA2 activity, and reduction of Din content. PMID- 10678153 TI - Scavenging effect of chinonin on free radicals studied with quantum chemistry. AB - AIM: To study whether the xanthonoid structure can enhance the ability of chinonin to scavenge free radicals and to understand the sequence of activity of chinonin hydroxyls. METHODS: Semiempirical quantum chemistry method AM1 was employed to calculate delta HOF values, the differences between heat of formations of mother molecule and its free radicals, and spin density distribution of different states of chinonin. delta HOF values were used as theoretical indices to measure scavenging activity of chinonin on free radicals and effects of structure on delta HOF values were investigated. RESULTS: delta HOF values of different phenolics were calculated to be 139.09 kJ.mol-1 (O5-H6), 141.46 kJ.mol-1 (O4-H5), 185.14 kJ.mol-1 (O2-H2) and 195.71 kJ.mol-1 (O1-H1). Spin density distribution of free radicals were obtained as well. CONCLUSION: Xanthonoid structure of chinonin made ring C to display high passive effect on ring B, which reduced scavenging activity of phenolics of ring B on free radicals. The sequence of activities of chinonin hydroxyls was O5-H6 > O4-H5 > O2 H2 > O1-H1. PMID- 10678154 TI - Water-retention effect of suberogorgin was due to secretion of antidiuretic hormone in rat. AB - AIM: To study the mechanism of antidiuretic effect of suberogorgin (Sub). METHODS: Conscious rat was given i.g. Sub 3.16 mg.kg-1 20 min after water-loaded treatment and then urine was collected in metabolic cage. Ion excretion was determined in atomic emission spectrometry. Urinary prostaglandin E (PGE), plasma PGE, antidiuretic hormone (ADH), and aldosterone were measured with RIA. Sub vs pituitrin or DOCA effects were carried out in hypophysectomized or adrenalectomized rats. RESULTS: The urine volume and the excretions of urinary sodium and potassium were decreased, maximally by 91%, 76%, and 86%, during the 24-h period after Sub. This antidiuretic effect possessed a progressive weakening with time. The concentrations of urinary PGE, plasma PGE, and ADH were increased by 25%, 212%, and 538%, respectively, but plasma aldosterone was not significantly influenced, 2 h after Sub dosing. The response of urine-excretion of rat to Sub was almost resisted by hypophysectomy but not by adrenalectomy. CONCLUSION: Sub decreased the urine excretion by, at least in part, accelerating the secretion of ADH but neither by PGE nor by aldosterone. PMID- 10678155 TI - Effect of aerobic exercise and ginsenosides on lipid metabolism in diet-induced hyperlipidemia mice. AB - AIM: To study the effect of aerobic exercise and its combination with Gin (ginsenosides from stems and leaves of ginseng) on lipid metabolism in diet induced hyperlipidemia mice. METHODS: The mouse hyperlipidemia model was set up by feeding high cholesterol diet. Unloaded swimming was designed to be a manner of aerobic exercise. The effects of aerobic exercise and its combination with Gin on total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-c) in serum, malondialdehyde (MDA), and superoxide dismutase (SOD) in liver tissue were measured; the thymus and liver were weighed. RESULTS: (1) The mouse hyperlipidemia model was set up successfully: TC and MDA increased (P < 0.05) but HDL-c and SOD decreased (P < 0.05); the liver weight increased and the thymus weight reduced; fatty liver was found; (2) aerobic exercise reduced TC but increased MDA and HDL-c in cholesterol-rich diet mice; the liver weight did not reduce, and fatty liver did not clear up; and (3) when aerobic exercise combined with Gin, TC and TG decreased markedly (P < 0.01), and MDA also decreased (P < 0.05); SOD and HDL-c increased markedly (P < 0.01); the thymus weight increased and the liver weight decreased to normal level; fatty liver cleared up. CONCLUSION: Aerobic exercise could lower serum lipid to some extent but could not satisfactorily regulate lipid metabolism. When combined with Gin, aerobic exercise could better lower serum lipid, regulate lipid metabolism, promote antioxidation, and enhance immune activity. PMID- 10678156 TI - [Effect of endothelin-1 injected into rostral ventrolateral medulla on cardiovascular responses in cats]. AB - AIM: To study the effect of endothelin-1 (ET-1) in the rostral ventrolateral medulla (rVLM) on cardiovascular responses in cats. METHODS: The stereotatic technique and microinjection method were used. RESULTS: ET-1 (4 mumol.L-1 0.5 microL) microinjected into rVLM induced mean arterial pressure (MAP) increasing (3.7 +/- 1.3) kPa, heart rate (HR) accelerating (29 +/- 7) beats.min-1, and renal nerve activity (RNA) intensifying 45% +/- 10%. The effects were dose-dependent. Before and after bilateral vagotomy, there was no significant difference in the reaction of MAP, HR, and RNA. After intravenous injection with phentolamine (5 mg.kg-1, alpha-blocker), ET-1 did not induce significant change of MAP. ET-1 raised the content of peripheral plasma argipressin (Arg) from (12.4 +/- 6.5) to (70.3 +/- 24.2) ng.L-1 with radioimmunoassay, and showed a correlation with MAP changes. ET-1 induced heart rhythm disorder (HRhD) in acute myocardiac ischemia, the occur time of HRhD was (4.8 +/- 2.9) min, and the score was 4.4 +/- 1.6, and it was significantly different from control. CONCLUSION: ET-1 microinjected into rVLM could involve with control regulation of cardiovascular and sympathetic nerve activity. PMID- 10678157 TI - Action of free radical in podophyllic acid piperindyl hydrazone nitroxide radical on its antitumor activity and toxicity. AB - AIM: To study the action of free radical in the spin-labeled podophyllotoxin derivative, podophyllic acid piperindyl hydrazone nitroxide radical (GP-1) on its antitumor activity and toxicity, by comparison with those of its free radical reduced product, podophyllic acid piperindyl hydrazone (GP-1-H). METHODS: After treatment with GP-1 and GP-1-H, the inhibitory effects on the growth of mouse transplantable tumors were determined; MTT formazan formation, [3H]deoxythymidine ([3H]TdR) incorporation, cell cycle progression, and mitotic index of SGC-7901 or L1210 cells were measured; the acute toxicity and immune function of mice were assayed. RESULTS: At doses of 1/6 and 1/12 LD50, the inhibitory rates against Lewis lung carcinoma were 60.3% and 42.1% (GP-1), 38.9% and 10.3% (GP-1-H), respectively; more effective antitumor activity of GP-1 against P388, HePS, and S 180 than that of GP-1-H were found. In vitro, GP-1 exhibited more powerful inhibitory effects on the proliferation and DNA synthesis of SGC-7901 and L1210 cells than GP-1-H. GP-1 and GP-1-H arrested the L1210 cells at G2/M phase with a corresponding decrease of the cells in G1 phase, and increased the mitotic index of the cells; but the effects of GP-1-H were weaker than those of GP-1. After treatment with doses of 1/4 and 1/8 LD50 for 5 d, no significant difference on immune function of mice between GP-1 and GP-1-H was found. CONCLUSION: GP-1 had more powerful antitumor activities than GP-1-H. The free radical in the spin labeled podophyllotoxin derivative, GP-1, played an important role in its antitumor activity. PMID- 10678158 TI - Emergency department evaluation and treatment of pelvic fractures. AB - Pelvic fractures are associated with a high morbidity and mortality rate. This article reviews the anatomy of the pelvis, discusses fracture patterns commonly seen in patients with an injured pelvis, and proposes a new method of classifying pelvic fractures based on potential associated injuries. Finally, algorithms for the management of hemodynamically stable and unstable patients with pelvic fractures are presented. PMID- 10678159 TI - Emergency department evaluation and treatment of hip and thigh injuries. AB - This article reviews the clinical and diagnostic evaluation of patients with injuries to the hip and thigh. The history and physical examination, appropriate imaging strategies, complications and associated injuries, analgesia, treatment, and appropriate patient disposition are emphasized. PMID- 10678160 TI - Emergency department evaluation and treatment of knee and leg injuries. AB - The knee is one of the most commonly injured joints in the human body, and, largely because of athletic injuries, they are increasing in frequency in the United States. This article provides a brief overview of knee anatomy, examines radiographic imaging techniques and arthrocentesis of the knee, and discusses injuries specific to the knee. An overview of leg anatomy is also presented, along with discussions of specific fractures common to the leg. PMID- 10678161 TI - Emergency department evaluation and treatment of ankle and foot injuries. AB - Ankle and foot injuries are among the most common sports injuries and extremity complaints presenting to the emergency department. Although generally benign, some of these injuries have prolonged morbidity. This article reviews the anatomy of the foot and ankle and examines the approach and therapy for common types of injuries. PMID- 10678162 TI - Soft tissue complications of orthopedic emergencies. AB - The emergency physician encounters a diversity of potentially devastating and disabling soft tissue maladies. This article reviews the literature and approach to the compartment syndrome and Volkmann contracture, reflex sympathetic dystrophy and causalgia, fracture blisters, and gas gangrene. PMID- 10678163 TI - Systemic analgesia and sedation in managing orthopedic emergencies. AB - Many potent agents have become available in the emergency department for providing systemic analgesia and sedation for painful orthopedic procedures. This article details the pharmacology and principles of systemic analgesia and sedation, which will help the emergency physician provide maximal patient comfort with minimal complications during painful procedures. The use of an appropriate agent in these situations will optimize the outcome of the procedure itself and result in greater patient satisfaction. PMID- 10678164 TI - The activation loop of phosphatidylinositol phosphate kinases determines signaling specificity. AB - Phosphatidylinositol-4,5-bisphosphate plays a pivotal role in the regulation of cell proliferation and survival, cytoskeletal reorganization, and membrane trafficking. However, little is known about the temporal and spatial regulation of its synthesis. Higher eukaryotic cells have the potential to use two distinct pathways for the generation of phosphatidylinositol-4,5-bisphosphate. These pathways require two classes of phosphatidylinositol phosphate kinases, termed type I and type II PIP kinases. While highly related by sequence, these kinases localize to different subcellular compartments, phosphorylate distinct substrates, and are functionally nonredundant. Here, we show that a 20- to 25 amino acid loop spanning the catalytic site, termed the activation loop, determines both enzymatic specificity and subcellular targeting of PIP kinases. Therefore, the activation loop controls signaling specificity and PIP kinase function at multiple levels. PMID- 10678165 TI - Diaphanous-related formins bridge Rho GTPase and Src tyrosine kinase signaling. AB - We have examined the role of the mouse Diaphanous-related formin (DRF) Rho GTPase binding proteins, mDia1 and mDia2, in cell regulation. The DRFs are required for cytokinesis, stress fiber formation, and transcriptional activation of the serum response factor (SRF). 'Activated' mDia1 and mDia2 variants, lacking their GTPase binding domains, cooperated with Rho-kinase or ROCK to form stress fibers but independently activated SRF. Src tyrosine kinase associated and co-localized with the DRFs in endosomes and in mid-bodies of dividing cells. Inhibition of Src also blocked cytokinesis, SRF induction by activated DRFs, and cooperative stress fiber formation with active ROCK. Our results show that the DRF proteins couple Rho and Src during signaling and the regulation of actin dynamics. PMID- 10678166 TI - Microtubule binding to Smads may regulate TGF beta activity. AB - Smad proteins are intracellular signaling effectors of the TGF beta superfamily. We show that endogenous Smad2, 3, and 4 bind microtubules (MTs) in several cell lines. Binding of Smads to MTs does not require TGF beta stimulation. TGF beta triggers dissociation from MTs, phosphorylation, and nuclear translocation of Smad2 and 3, with consequent activation of transcription in CCL64 cells. Destabilization of the MT network by nocodazole, colchicine, or a tubulin mutant disrupts the complex between Smads and MTs and increases TGF beta-induced Smad2 phosphorylation and transcriptional response in CCL64 cells. These data demonstrate that MTs may serve as a cytoplasmic sequestering network for Smads, controlling Smad2 association with and phosphorylation by activated TGF beta receptor I, and suggest a novel mechanism for the MT network to negatively regulate TGF beta function. PMID- 10678167 TI - Left-right asymmetric expression of lefty2 and nodal is induced by a signaling pathway that includes the transcription factor FAST2. AB - The left-right (L-R) asymmetric expression of lefty2 and nodal is controlled by a left side-specific enhancer (ASE). The transcription factor FAST2, which can mediate signaling by TGF beta and activin, has now been identified as a protein that binds to a conserved sequence in ASE. These FAST2 binding sites were both essential and sufficient for L-R asymmetric gene expression. The Fast2 gene is bilaterally expressed when nodal and lefty2 are expressed on the left side. TGF beta and activin can activate the ASE activity in a FAST2-dependent manner, while Nodal can do so in the presence of an EGF-CFC protein. These results suggest that the asymmetric expression of lefty2 and nodal is induced by a left side-specific TGF beta-related factor, which is most likely Nodal itself. PMID- 10678168 TI - Signal transduction by a death signal peptide: uncovering the mechanism of bacterial killing by penicillin. AB - The binding of bactericidal antibiotics like penicillins, cephalosporins, and glycopeptides to their bacterial targets stops bacterial growth but does not directly cause cell death. A second process arising from the bacteria itself is necessary to trigger endogenous suicidal enzymes that dissolve the cell wall during autolysis. The signal and the trigger pathway for this event are completely unknown. Using S. pneumoniae as a model, we demonstrate that signal transduction via the two-component system VncR/S triggers multiple death pathways. We show that the signal sensed by VncR/S is a secreted peptide, Pep27, that initiates the cell death program. These data depict a novel model for the control of bacterial cell death. PMID- 10678169 TI - Hedgehog creates a gradient of DPP activity in Drosophila wing imaginal discs. AB - Hedgehog (HH) and Decapentaplegic (DPP) direct anteroposterior patterning in the developing Drosophila wing by functioning as short- and long-range morphogens, respectively. Here, we show that the activity of DPP is graded and is directly regulated by a novel HH-dependent mechanism. DPP activity was monitored by visualizing the activated form of Mothers against dpp (MAD), a cytoplasmic transducer of DPP signaling. We found that activated MAD levels are highest near the source of DPP but are unexpectedly low in the cells that express dpp. HH induces dpp in these cells; it also attenuates their response to DPP by downregulating expression of the DPP receptor thick veins (tkv). We suggest that regulation of tkv by HH is a key part of the mechanism that controls the level and distribution of DPP. PMID- 10678170 TI - The murine SCP3 gene is required for synaptonemal complex assembly, chromosome synapsis, and male fertility. AB - During meiosis, the homologous chromosomes pair and recombine. An evolutionarily conserved protein structure, the synaptonemal complex (SC), is located along the paired meiotic chromosomes. We have studied the function of a structural component in the axial/lateral element of the SC, the synaptonemal complex protein 3 (SCP3). A null mutation in the SCP3 gene was generated, and we noted that homozygous mutant males were sterile due to massive apoptotic cell death during meiotic prophase. The SCP3-deficient male mice failed to form axial/lateral elements and SCs, and the chromosomes in the mutant spermatocytes did not synapse. While the absence of SCP3 affected the nuclear distribution of DNA repair and recombination proteins (Rad51 and RPA), as well as synaptonemal complex protein 1 (SCP1), a residual chromatin organization remained in the mutant meiotic cells. PMID- 10678171 TI - CDK inactivation is the only essential function of the APC/C and the mitotic exit network proteins for origin resetting during mitosis. AB - Passage through mitosis is required to reset replication origins for the subsequent S phase. During mitosis, a series of biochemical reactions involving cyclin-dependent kinases (CDKs), the anaphase promoting complex or cyclosome (APC/C), and a mitotic exit network including Cdc5, 14, and 15 coordinates the proper separation and segregation of sister chromatids. Here we show that cyclin B/CDK inactivation can drive origin resetting in either early S phase or mitosis. This origin resetting occurs efficiently in the absence of APC/C function and mitotic exit network function. We conclude that CDK inactivation is the single essential event in mitosis required to allow pre-RC assembly for the next cell cycle. PMID- 10678172 TI - Identification of two catalytic residues in RAG1 that define a single active site within the RAG1/RAG2 protein complex. AB - During V(D)J recombination, the RAG1 and RAG2 proteins cooperate to catalyze a series of DNA bond breakage and strand transfer reactions. The structure, location, and number of active sites involved in RAG-mediated catalysis have as yet not been determined. Using protein secondary structure prediction algorithms, we have identified a region of RAG1 with possible structural similarities to the active site regions of transposases and retroviral integrases. Based on this information, we have identified two aspartic acid residues in RAG1 (D600 and D708) that function specifically in catalysis. The results support a model in which RAG1 contains a single, divalent metal ion binding active site structurally related to the active sites of transposases/integrases and responsible for all catalytic functions of the RAG protein complex. PMID- 10678173 TI - The eIF1A solution structure reveals a large RNA-binding surface important for scanning function. AB - The translation initiation factor eIF1A is necessary for directing the 43S preinitiation complex from the 5' end of the mRNA to the initiation codon in a process termed scanning. We have determined the solution structure of human eIF1A, which reveals an oligonucleotide-binding (OB) fold and an additional domain. NMR titration experiments showed that eIF1A binds single-stranded RNA oligonucleotides in a site-specific, but non-sequence-specific manner, hinting at an mRNA interaction rather than specific rRNA or tRNA binding. The RNA binding surface extends over a large area covering the canonical OB fold binding site as well as a groove leading to the second domain. Site-directed mutations at multiple positions along the RNA-binding surface were defective in the ability to properly assemble preinitiation complexes at the AUG codon in vitro. PMID- 10678174 TI - Mammalian microsomal cytochrome P450 monooxygenase: structural adaptations for membrane binding and functional diversity. AB - Microsomal cytochrome P450s participate in xenobiotic detoxification, procarcinogen activation, and steroid hormone synthesis. The first structure of a mammalian microsomal P450 suggests that the association of P450s with the endoplasmic reticulum involves a hydrophobic surface of the protein formed by noncontiguous portions of the polypeptide chain. This interaction places the entrance of the putative substrate access channel in or near the membrane and orients the face of the protein proximal to the heme cofactor perpendicular to the plane of the membrane for interaction with the P450 reductase. This structure offers a template for modeling other mammalian P450s and should aid drug discovery and the prediction of drug-drug interactions. PMID- 10678175 TI - Mapping interactions between nuclear transport factors in living cells reveals pathways through the nuclear pore complex. AB - The interactions between transport receptors and proteins of the nuclear pore complex (NPC) are fundamental to understanding nucleocytoplasmic transport. In order to delineate the path that a particular transport receptor takes through the NPC, we have employed fluorescence resonance energy transfer (FRET) between enhanced cyan and yellow fluorescent proteins (ECFP, EYFP) in living cells. A panel of yeast strains expressing functional receptor--ECFP and nucleoporin--EYFP fusions has been analyzed with a FRET assay. With this approach, we define points of contact in the NPC for the related importin Pse1/Kap121 and exportin Msn5. These data demonstrate the utility of FRET in mapping dynamic protein interactions in a genetic system. Furthermore, the data indicate that an importin and exportin have overlapping pathways through the NPC. PMID- 10678176 TI - Disruption of the beta-sarcoglycan gene reveals pathogenetic complexity of limb girdle muscular dystrophy type 2E. AB - Limb-girdle muscular dystrophy type 2E (LGMD 2E) is caused by mutations in the beta-sarcoglycan gene, which is expressed in skeletal, cardiac, and smooth muscle. beta-sarcoglycan-deficient (Sgcb-null) mice developed severe muscular dystrophy and cardiomyopathy with focal areas of necrosis. The sarcoglycan sarcospan and dystroglycan complexes were disrupted in skeletal, cardiac, and smooth muscle membranes. epsilon-sarcoglycan was also reduced in membrane preparations of striated and smooth muscle. Loss of the sarcoglycan-sarcospan complex in vascular smooth muscle resulted in vascular irregularities in heart, diaphragm, and kidneys. Further biochemical characterization suggested the presence of a distinct epsilon-sarcoglycan complex in skeletal muscle that was disrupted in Sgcb-null mice. Thus, perturbation of vascular function together with disruption of the epsilon-sarcoglycan-containing complex represents a novel mechanism in the pathogenesis of LGMD 2E. PMID- 10678177 TI - The ves multigene family of B. bovis encodes components of rapid antigenic variation at the infected erythrocyte surface. AB - B. bovis, an intraerythrocytic protozoal parasite, establishes chronic infections in cattle in part through rapid variation of the polymorphic, heterodimeric VESA1 protein on the infected erythrocyte surface and sequestration of mature parasites. We describe the characterization of the ves1 alpha gene encoding the VESA1a subunit, thus providing a description of a gene whose product is involved in rapid antigenic variation in a babesial parasite. This three-exon gene, a member of a multigene family (ves), encodes a polypeptide with no cleavable signal sequence, a single predicted transmembrane segment, and a cysteine/lysine rich domain. Variation appears to involve creation and modification or loss of a novel, transcribed copy of the gene. PMID- 10678178 TI - Rnq1: an epigenetic modifier of protein function in yeast. AB - Two protein-based genetic elements (prions) have been identified in yeast. It is not clear whether other prions exist, nor is it understood how one might find them. We established criteria for searching protein databases for prion candidates and found several. The first examined, Rnq1, exists in distinct, heritable physical states, soluble and insoluble. The insoluble state is dominant and transmitted between cells through the cytoplasm. When the prion-like region of Rnq1 was substituted for the prion domain of Sup35, the protein determinant of the prion [PSI+], the phenotypic and epigenetic behavior of [PSI+] was fully recapitulated. These findings identity Rnq1 as a prion, demonstrate that prion domains are modular and transferable, and establish a paradigm for identifying and characterizing novel prions. PMID- 10678179 TI - The crystal structure of the nuclear receptor for vitamin D bound to its natural ligand. AB - The action of 1 alpha, 25-dihydroxyvitamin D3 is mediated by its nuclear receptor (VDR), a ligand-dependent transcription regulator. We report the 1.8 A resolution crystal structure of the complex between a VDR ligand-binding domain (LBD) construct lacking the highly variable VDR-specific insertion domain and vitamin D. The construct exhibits the same binding affinity for vitamin D and transactivation ability as the wild-type protein, showing that the N-terminal part of the LBD is essential for its structural and functional integrity while the large insertion peptide is dispensable. The structure reveals the active conformation of the bound ligand and allows understanding of the different binding properties of some synthetic analogs. PMID- 10678180 TI - VASA mediates translation through interaction with a Drosophila yIF2 homolog. AB - The Drosophila gene vasa (vas) encodes an RNA-binding protein required for embryonic patterning and germ cell specification. In vas mutants, translation of several germline mRNAs is reduced. Here we show that VAS interacts directly with the Drosophila homolog of yeast translation initiation factor 2, encoded by a novel gene, dIF2. Embryos produced by vas/+; dIF2/+ females have pattern defects and fewer germline progenitor cells, indicating a functional interaction between endogenous vas and dIF2 activities. Mutations in other translation initiation factors do not enhance the vas phenotype, suggesting that dIF2 has a particular role in germ plasm function. We conclude that VAS regulates translation of germline mRNAs by specific interaction with dIF2, an essential factor conserved from bacteria to humans. PMID- 10678181 TI - Nf1 and Gmcsf interact in myeloid leukemogenesis. AB - The NF1 tumor suppressor gene encodes neurofibromin, a GTPase-activating protein (GAP) for p21ras (Ras). Children with NF1 are predisposed to juvenile myelomonocytic leukemia (JMML). Some heterozygous Nf1 mutant mice develop a similar myeloproliferative disorder (MPD), and adoptive transfer of Nf1-deficient fetal liver cells consistently induces this MPD. Human JMML and murine Nf1 deficient cells are hypersensitive to granulocyte-macrophage colony-stimulating factor (GM-CSF) in methylcellulose cultures. We generated hematopoietic cells deficient in both Nf1 and Gmcsf to test whether GM-CSF is required to drive excessive proliferation of Nf1-/- cells in vivo. Here we show that GM-CSF play a central role in establishing and maintaining the MPD and that recipients engrafted with Nf1-/- Gmcsf-/- hematopoietic cells are hypersensitive to exogenous GM-CSF. PMID- 10678182 TI - Disclosure of HIV status and its consequences. PMID- 10678183 TI - HIV seropositivity in the Western Cape. PMID- 10678184 TI - Chemically dependent medical practitioners--the prognosis for treatment. PMID- 10678185 TI - Durban Malaria Conference--a short report. PMID- 10678186 TI - Rationalizing the use of intravenous therapy in newborns. PMID- 10678187 TI - AZT--the saga continues. PMID- 10678188 TI - Saving mothers--report on maternal deaths in South Africa. PMID- 10678190 TI - R100M Vincent Pallotti Hospital expansion completed PMID- 10678189 TI - SAMA assists with development of health package for regional hospitals. PMID- 10678192 TI - Health and human rights in post-apartheid South Africa. PMID- 10678191 TI - Clinical progression of HIV infection in adults. PMID- 10678193 TI - The value of electromyography and nerve conduction studies in the diagnosis of neurological disorders. PMID- 10678194 TI - How iron should be administered. PMID- 10678195 TI - Benzoylserine and benzoylthreonine in propionic acidaemia treated with sodium benzoate. PMID- 10678196 TI - Microbiologically proven Pneumocystis carinii pneumonia (PCP) at Ga-Rankuwa Hospital. PMID- 10678197 TI - Growth and nutrition in South African children with cystic fibrosis. AB - OBJECTIVES: To study nutritional status and its dietary correlates in a South African cystic fibrosis (CF) population. DESIGN: Cross-sectional survey. POPULATION: Thirty-eight children and adolescents attending the CF clinic at Red Cross War Memorial Children's Hospital, Cape Town. METHODS: Standard anthropometry and a 3-day weighted food record. RESULTS: Median percentage expected weight for height (WFH) was 93 (interquartile range 84-101). Sixteen per cent of patients were below the 5th percentile for height. The proportion of patients who were malnourished (WFH less than 90) was greater among those over 10 years of age (47% v. 14.3%, chi 2 = 4.33, P = 0.037). Sixty-eight per cent of patients consumed less than the recommended daily intake of energy. There was no correlation between WFH and energy intake. Fat intake represented 29.6% (interquartile range 27.5-33%) of daily energy intake. CONCLUSIONS: Young South African children with CF are growing well despite relatively low intakes of energy and fat. Greater attention needs to be given to overcoming malnutrition among older children. PMID- 10678198 TI - Pregnancy- and lactation-related folate deficiency in South Africa--a case for folate food fortification. AB - OBJECTIVE: Characterisation of patients presenting with megaloblastic anaemia according to clinical, sociological, haematological and aetiological aspects of their disease, and use of these findings to increase awareness among clinicians and to make recommendations regarding changes in national health policy. METHODS: This study included 104 patients presenting with megaloblastic anaemia to a large referral hospital over a 1-year period. Data were collected and analysed in terms of age, gender, parity, gravidity, duration of lactation, socio-economic status, geographical origins, diet, previous haematinic treatment, clinical presentation and haematological measurements. RESULTS: The most common cause of megaloblastic anaemia was pernicious anaemia or probable pernicious anaemia (50%), followed by pregnancy- and lactation-related folate deficiency (32%); of these patients, the majority (28) presented postpartum while lactating; 5 patients were in the immediate puerperal period of 6 weeks, and a further 16 were seen during the first year and 7 during the second year following delivery. Only 4 patients were pregnant, and it is noteworthy that 2 of these were still lactating at 34 weeks' gestation. CONCLUSION: Pregnancy- and lactation-related folate deficiency up to 2 years after delivery remains a common cause of megaloblastic anaemia in South Africa. Certain communities in rural South Africa have recently been shown to have high incidences of both neural tube defects and folate deficiency. The fortification of a staple food (e.g. maize or flour) with folic acid is feasible, inexpensive, safe and likely to be beneficial. This practice should reduce the prevalences of megaloblastic anaemia in fertile women, neural tube defects, other congenital abnormalities, intra-uterine growth retardation, prematurity and possibly cardiovascular disease. There is urgent need for a national policy in this regard. PMID- 10678199 TI - Intracranial mass lesions in HIV-positive patients--the KwaZulu/Natal experience. Neuroscience AIDS Research Group. AB - BACKGROUND: Neurological disease heralds the development of AIDS in 10-20% of HIV seropositive individuals. In over half of these cases the presentation will be that of an intracranial mass lesion (IML). In developed countries toxoplasmosis is the most frequent cause of IML in a positive patient, followed by primary central nervous system lymphoma. Less common causes include tuberculomas, cryptococcomas, abscesses and gummas. As a result of these observations, the algorithm adopted in developed countries calls for initial empirical treatment for toxoplasmosis. Biopsy of the IML is only considered if there is no response to treatment after 10-14 days. Whether such an algorithm would be applicable to the local population is unknown. OBJECTIVE: We undertook a prospective study to determine the type and frequency of IML in local HIV-seropositive patients. A secondary objective, based on the findings, was to develop a local algorithm of management. PATIENTS AND METHODS: Over a 17-month period HIV-seropositive individuals with an IML were entered into the study. Biopsy or aspiration of the lesion was performed either stereotactically or free-hand. Tissue obtained was processed for routine and special histological studies. RESULTS: In the 38 cases where tissue was obtained, the most frequent cause of the IML was toxoplasmosis followed by encephalitis of obscure origin', brain abscess and tuberculoma/mycobacterial infection. CONCLUSION: This study demonstrated that the spectrum of IML seen locally was similar to that in developed countries. The management protocol used elsewhere was therefore adopted for local patients. PMID- 10678201 TI - Medical recipients of the Victoria Cross during the Anglo-Boer War, 1899-1902. PMID- 10678200 TI - Aorto-iliac occlusive disease in the different population groups--clinical pattern, risk profile and results of reconstruction. AB - BACKGROUND: It has previously been accepted that atherosclerotic disease is uncommon among blacks worldwide; however, recent studies have increasingly reported atherosclerotic disease in this group. STUDY DESIGN: Prospective study of hospital patients with aorto-iliac occlusive disease presenting to the vascular service of the Durban metropolitan hospitals. The study was designed to assess clinical pattern, risk profile and results of reconstruction in these patients. METHODS: This is a study of 688 patients with aorto-iliac occlusive disease managed over 9 years in Durban, with clinical pattern and risk factors compared in the different population groups. A subgroup of 492 patients underwent aortobifemoral bypass, providing material for comparison of the results of reconstruction in the different population groups. RESULTS: More black patients presented with gangrene and threatened limb, whereas whites tended to present early with claudication. All groups had hypertension and diabetes as risk factors. In addition, whites and Indians had ischaemic heart disease, which was not found among blacks. Mortality was 5% (blacks 1.8%, whites 8.5%, Indians 5%). Medium-term occlusion rates were 19% in blacks, 13% in Indians and 5% among whites. Five-year cumulative patency rates were 92% for whites, 77% for Indians and 74% for blacks. CONCLUSION: Whites do significantly better than blacks, who tend to present at an advanced stage of the disease. The presence of ischaemic heart disease among whites and Indians contributes to the higher mortality in these groups. PMID- 10678202 TI - Media events. PMID- 10678203 TI - A statistical perspective on gender in medicine. PMID- 10678204 TI - Gender bias in biomedical research. PMID- 10678205 TI - Gender bias in medical education: twenty vignettes and recommended responses. PMID- 10678206 TI - Gender discrimination: a health and career development problem for women physicians. PMID- 10678208 TI - The need for a national cancer control strategy in New Zealand. PMID- 10678207 TI - Gender bias in a peer-reviewed medical journal. PMID- 10678209 TI - Socio-demographic characteristics of New Zealand smokers: results from the 1996 census. AB - AIMS: To examine the key socio-demographic characteristics of adult smokers in New Zealand based on 1996 census data. METHODS: Data were obtained from the 1996 Census of Populations and Dwellings on smoking status and key socio-demographic variables. Age standardised smoking prevalence rates were calculated. RESULTS: Of the New Zealand population aged 15 years and over, 23.7% reported that they were regular smokers. Maori have the highest smoking prevalence in New Zealand (40.5%) and the peak rate is 55% among young Maori women aged 25-29 years. Pacific Island people, particularly males, have higher smoking rates than Europeans, while among Asians, the rate for males is three to four times the rate for females. People with no qualifications, who are unemployed or earning less than $30,000 per year, and women with high parities also have relatively high smoking prevalence rates. CONCLUSION: The smoking rates of New Zealand population groups are highly heterogeneous and there is substantial scope for focusing tobacco control interventions on those groups with the highest prevalence. PMID- 10678210 TI - How many pregnant women in Christchurch are using folic acid supplements in early pregnancy? AB - AIMS: To determine the proportion of pregnant women in Christchurch using folic acid supplements in early pregnancy. To evaluate the level of current knowledge relating to folic acid amongst pregnant women. To determine the main sources from which this information was gained. METHODS: A short questionnaire was administered to 191 pregnant women in Christchurch during antenatal visits with their lead maternity carer. The survey contained questions relating to knowledge about folic acid and use together with sources of information regarding folic acid. Obstetric and demographic details were also collected. RESULTS: The response rate was 95.5%. Ninety-one per cent (174/191) of participants had heard of folic acid and, of these, 63% knew that folic acid reduces the risk of spina bifida. Of the 191 participants in the study, 118(62%) took folic acid supplements at some stage of their pregnancy, however, only 33(17%) had taken periconceptual folic acid supplements. Of the 44% of all women in the study with a planned pregnancy, only 35% had taken folic acid supplements periconceptually. Of those women with an unplanned pregnancy (55%), only 2.8% had taken a folic acid supplement periconceptually. The main sources of advice for women relating to folic acid were general practitioners (48%) or media advertising, either in the form of a magazine, or health pamphlet or television promotion (20%). CONCLUSIONS: The results of this study indicate that the level of knowledge amongst women of child-bearing age relating to folic acid is relatively high compared with other countries. Despite this high level of knowledge, only a small percentage of women are actually consuming a folic acid supplement during the recommended periconceptual period due in part to the high proportion of unplanned pregnancies. These results emphasize the need for an effective public health strategy to ensure that all women of child-bearing age have access to an adequate folic acid intake. PMID- 10678211 TI - Outcomes of transrectal ultrasound scan of the prostate with sector biopsies for 323 New Zealand men with suspicion of prostate cancer. AB - AIMS: To assess the results and clinical outcomes of the first four years of transrectal ultrasound scanning (TRUS) with sector biopsies of the prostate, as the definitive second-line investigation for men with suspicion of prostate cancer, including comparability with subsequent information from histology of surgical specimens. METHODS: Information was collated from the author's ongoing surgical audit. TRUS and sector biopsies were carried out as a rooms procedure using a Toshiba Sono-Layer SSA-270A ultrasound machine with a PVL 725 RT transrectal probe and biopsy guide. Six or eight sector biopsies were taken with a Manan biopsy gun using 18 French gauge biopsy needles. Prophylactic ciprofloxacin and tinidazole were administered. Men with suspicion of prostate specific antigen assay or digital rectal examination were considered for the investigation, especially if they were candidates for potentially curative treatments. RESULTS: Of 330 TRUS procedures performed on 323 men, 328 were done in rooms under local anaesthetic. The only significant complication was a transient bacteraemia in one patient not taking the antibiotics. Twenty men had prostate intraepithelial neoplasia or atypia, and 94 (29%) had cancer, of whom 24 (25%) had evidence of metastases. Curative treatment by radical prostatectomy or radiotherapy was attempted in 62 men (66%) and 28 are being managed by surveillance only. Comparisons with subsequent radical prostatectomy in 44 men showed that if only one TRUS biopsy core was involved with cancer (15 men), surgical margins were clear. For those with more cores involved (29 men), one third (9 men) had positive margins or capsular perforation. The Gleason Score was different, more often higher, for 29 men by histology at radical prostatectomy compared to TRUS. Twenty-eight men had a subsequent transurethral resection of the prostate of which the histology resulted in management changes for ten of the 28, including three who were placed on surveillance and seven who underwent radical prostatectomy. CONCLUSIONS: TRUS and sector biopsy is a tolerable rooms procedure for men with suspician of prostate cancer. Complications are rare if prophylactic antibiotics are taken. The procedure provides information of sufficient quality to advise patients further. No defects in specificity were detected. Defects in sensitivity were demonstrated by comparison with later histology following transurethral resection. Therefore, men with benign results at TRUS need ongoing follow-up. TRUS histology tends to underestimate the extent of cancer present as determined by subsequent radical prostatectomy histology. When used with prostate-specific antigen and digital rectal examination, TRUS and sector biopsy is capable of detecting prostate cancer before it has metastasized, more reliably than if symptoms are awaited before diagnosis is attempted. It may also recognize forms of cancer of apparent low clinical significance which can be managed by surveillance. PMID- 10678212 TI - For whom is the Caesarean section rate high? AB - AIM: To define a method for examining and comparing Caesarean section rates. METHODS: Data on Caesarean section rates at National Women's Hospital for 1997 were analysed by two methods which adjust Caesarean section rates according to casemix: 1. standard nulliparae and 2. grouping women by factors influencing Caesarean section rates. RESULTS: The Caesarean section rate in New Zealand is rising and National Women's Hospital had a Caesarean section rate of 22.3% in 1997. The instrumental vaginal delivery rate was 13.1%. Our 1997 data were analysed for casemix -- standard nulliparae and dividing women into six distinct groups. The Caesarean section rate for standard nulliparae was 19.2%. Nulliparae had an increased induction of labour rate compared to multiparae (29.1% versus 22.9%, p < 0.001). The Caesarean section rate was increased in association with induction of labour for nulipare (26.0% versus 13.1%, p < 0.001), multiparae with unscarred uteri, (5.4% versus 2.5%, p < 0.001) and primiparae with previous caesareans (36.8% versus 30.4%, p = 0.02) CONCLUSION: We suggest our second method of grouping women by the most important factors influencing intervention rates should be used by obstetric units for benchmarking and internal audit purposes. PMID- 10678213 TI - Attention is needed to the high prevalence of vitamin D deficiency in our older population. AB - The aim of this study was to assess the prevalence of vitamin D deficiency in females residing in one large, aged-care facility in Auckland. Thirty-nine residents, most of whom were able to go outdoors without assistance, were tested for midwinter 25-hydroxyvitamin D, and, of these, 36 were tested again in midsummer. The prevalence of frank hypovitaminosis (<10 microg/L) was found to be 49% in midwinter and 33% in midsummer. The vitamin D status of such at-risk individuals could be normalised either by 15-30 minutes of daily sun exposure or, alternatively, a programme of supplementation. PMID- 10678214 TI - Patient enrollment with co-payments: implications for patient choice in general practice. PMID- 10678215 TI - Antibiotics in otitis media. PMID- 10678216 TI - Home birth outcome: a personal series. PMID- 10678217 TI - Screening for prothrombotic diathesis in family planning clinics. PMID- 10678218 TI - Medical Council and penalties for misconduct. PMID- 10678219 TI - Ethics of ACC continued. PMID- 10678220 TI - Mycobacterium xenopi lung infection. PMID- 10678221 TI - Measuring health-related quality of life using the SF-36. PMID- 10678222 TI - Medical practitioner guilty of misconduct. PMID- 10678223 TI - Cardiac allograft vasculopathy: the green lane hospital experience 1987-1998. AB - AIMS: To determine the prevalence of cardiac allograft vasculopathy in heart transplant recipients at Green Lane Hospital and to examine potential risk factors for vasculopathy. METHODS: We retrospectively reviewed the coronary angiograms of all cardiac transplant recipients. Angiography was usually performed one, two and five years after operation. The diagnosis of allograft vasculopathy was made if there was any evidence of coronary artery disease. Patients' medical records were reviewed for potential risk factors. RESULTS: Ninety-one patients underwent cardiac transplantation between December 1987 and March 1998. One year survival was 87%. Angiographic evidence of coronary disease was present in 30 patients and in three patients coronary lesions were first identified at post mortem. Vasculopathy was present in 25%, 35% and 61% of patients at one two and five years following transplant. Donor-acquired lesions could not be excluded as few patients had immediate postoperative angiograms for comparison. Five late deaths have been due to vasculopathy. Recipient age, race, donor age and ischaemic time were similar for those with and without vasculopathy. Frequency or severity of acute rejection episodes, cytomegalovirus infection, lipid profiles, diabetes and hypertension were not significantly different in patients with vasculopathy. CONCLUSION: Cardiac allograft vasculopathy is a common finding after heart transplantation. No definite risk factors were identified in this patient group. PMID- 10678224 TI - Postoperative adhesive small bowel obstruction: the resources impacts. AB - AIM: To assess the resource implications of managing small bowel obstruction, which is common and has diverse causes and outcomes. METHOD: A retrospective study of 332 patients documented to have postoperative, adhesive small bowel obstruction, from 1988 to 1996, was carried out. Complications and resources used were recorded and costs were determined. Results. There were 207 females and 125 males, with a median age of 63 years. There were 374 hospital admissions, in 121 (32.4%) of which no surgical operation was performed. Patients had a median hospital stay of eight days. The overall median cost for a patient who had no operation was NZ$1 039 (minimum $94, maximum $13 262), compared to NZ$7 630 (minimum $2 038, maximum $135 173) for a patient who had an operation. Postoperative adhesive small bowel obstruction accounted for 1.3% of all admissions, 59.2% of all cases of bowel obstruction, 65.2% of all admissions with small bowel conditions, 73.5% of laparotomies for bowel obstruction and 4.1% of all laparotomies. Sixty-eight patients (20.5%) developed a total of 102 complications and there were eight deaths (2.4%). CONCLUSION: Postoperative adhesive small bowel obstruction is a common condition, which is associated with a substantial morbidity and workload. The treatment of these conditions has significant health care costs. Most are emergencies and the costs of their management are very variable. PMID- 10678225 TI - Susceptibility of anaerobic bacteria in Auckland: 1991-1996. AB - AIM: To determine the antimicrobial susceptibility of local anaerobic bacteria. METHOD: The antimicrobial susceptibility of 357 obligate anaerobes collected between 1991 and 1997 was determined by a standard agar dilution method. Isolates tested included Bacteroides spp. 131, Fusobacterium spp. 12, Prevotella spp. 13, Veillonella spp. 5, Clostridium perfringens 27, other Clostridium spp. 29, Propionibacterium spp. 57, Actinomyces spp. 7, other non-sporing gram-positive bacilli 28 and Peptostreptococcus spp. 48. Ten antimicrobials were tested: penicillin, amoxycillin/ clavulanic acid, pipercillin/tazobactam, ceftriaxone, cefoxitin, cefotetan, imipenem, meropenem, clindamycin and metronidazole. RESULTS: Imipenem, pipercillin/tazobactam, meropenem and amoxycillin/clavulanic acid were active against virtually all anaerobes tested. Metronidazole was active against all anaerobic gram-negative bacteria and Clostridium spp., but had variable activity against other anaerobes. Cefoxitin was the most active cephalosporin against Bacteroides spp., with 76%, 64% and 15% of Bacteroides spp. being susceptible to cefoxitin, cefotetan and ceftriaxone, respectively. Penicillin had poor activity against anaerobic gram negative bacilli. Actinomyces and Propionibacterium spp. were susceptible to all antimicrobials tested except metronidazole. Variable results were obtained with other antimicrobial-organism combinations. Comparison of results with data from a previously published survey showed little change in susceptibility except for increased resistance of Bacteroides fragilis to ceftriaxone and Clostridium species (not C perfringens) to clindamycin. CONCLUSION: Our results update the local susceptibility profile of anaerobic bacteria and may be considered when choosing an antimicrobial agent for prophylaxis or treatment of anaerobic infections. PMID- 10678226 TI - The health of elderly residents in long term care institutions in New Zealand. AB - AIMS: To estimate the morbidity of residents in long term care institutions in Hamilton, New Zealand and examine the prescribing practice within these institutions. METHODS: One hundred residents were selected at random from the total population of residents in long-term institutions in Hamilton. Residents were examined by a physician to arrive at ICD-9 diagnoses and details were recorded about prescribing practice. RESULTS: Eighty residents were examined. Of these, 73% had dementia. Forty-five per cent were diagnosed as having heart failure and 44% cerebrovascular disease. The most common previously undiagnosed disorder was postural hypotension (24%). Residents were prescribed an average of 4.5 non-psychotropic medications. Seventy-seven per cent of residents were prescribed three or more medications. CONCLUSION: Elderly residents in long term care institutions in Hamilton have complex health care needs which offer particular challenges for doctors and other health care workers. Regular health care reviews could lead to more accurate diagnosis and better prescribing practice. PMID- 10678227 TI - Death due to unrecognised myocardial infarction causing left ventricular rupture: can we improve the diagnostic rate? AB - AIM: To investigate the clinical presentations of ruptured myocardial infarction, where the initial diagnosis of myocardial infarction was missed, to enhance the diagnostic rate of primary care physicians. METHODS: We studied 67 cases of myocardial infarction, terminating with left ventricular rupture, between January 1988 and December 1996. The study was restricted to sudden death where, at coroner-directed autopsy, a ruptured myocardial infarction was determined as the cause of death. It was also restricted to patients who consulted a doctor within the two weeks prior to death. The report made to the coroner by attending police and the autopsy report was studied, and the requisite data were abstracted. RESULTS: Half of our study group did not present with chest pain. Of the atypical presentations: 15/67 cases (22%) were from referred pain (neck, arm, abdomen or back), 12/ 67 patients presented with "flu-like illness" (18%), 4/67 cases had respiratory presentations (cough or shortness of breath) (6%) and 2/67 falls (3%). Of those with chest pain, 16/34 (47%) were diagnosed or referred and 2/15 infarcts with atypical or referred pain were diagnosed. None of those presenting with "flu like illness" or respiratory symptoms was diagnosed or referred. CONCLUSION: Fifty per cent of our patients had "silent" myocardial infarcts. A large proportion of this group complained of a flu-like illness, which is currently not considered a presentation of this disease. Patients at higher risk of a myocardial infarct, should be treated with a high index of suspicion when unwell, especially when complaining of a flu-like illness. Pathologically, posterior and lateral infarcts accounted for over half the cases. PMID- 10678228 TI - Screening for hepatitis B carriers: evidence and policy development in New Zealand. PMID- 10678229 TI - PHARMAC: at what cost? PMID- 10678230 TI - PHARMAC and good science? PMID- 10678231 TI - PHARMAC complicates treatment of glaucoma. PMID- 10678232 TI - Measuring health-related quality of life. PMID- 10678233 TI - Systemic inflammatory response syndrome (SIRS) from acute polyarticular gout. PMID- 10678234 TI - Diabetes complication screening. PMID- 10678235 TI - A changing world- the need and value of home visits. PMID- 10678236 TI - Do referring doctors and rheumatologists agree? PMID- 10678237 TI - Oral contraceptives and venous thromboembolism. PMID- 10678238 TI - Simple technology use in patient care at a distance. PMID- 10678239 TI - Radiologist found guilty of misconduct. PMID- 10678240 TI - Hyperhomocysteinaemia: a risk factor for vascular disease. PMID- 10678241 TI - Homocysteine levels in healthy New Zealanders and those with vascular disease. AB - AIMS: Levels of plasma homocysteine (tHcy) have been shown to vary between populations. The aim of the present study was to determine tHcy levels in a New Zealand population to facilitate interpretation of international reference ranges. METHODS: Fasting tHcy levels were determined in 431 volunteer men and women, aged between 17 and 83 years, from the greater Otago region. Subjects with self-reported incidences of vascular disease (n=138) were compared with healthy control subjects (n=293). RESULTS: Mean (SD) fasting tHcy level of the entire population was 8.3(3.5) micromol/L and men had significantly higher levels than women (9.0(3.4) and 7.8(3.5) micromol/L, respectively, p<0.05). Levels increased significantly with increasing age (0.5 micromol/L for every ten years). There was a small but non-significant difference in tHcy levels between subjects with and without vascular disease (difference after excluding two outliers and adjusting for age and gender; 0.63, 95% CI, -0.03,1.29). tHcy levels were not significantly correlated with lipid or lipoprotein levels. CONCLUSION: Levels of tHcy in a group of volunteers from the greater Otago region were similar to those reported in other populations at high risk of cardiovascular disease. Reference ranges derived from these populations would appear to be applicable for New Zealanders. tHcy measurements should be made when assessing individuals at high risk of vascular disease and intervention strategies considered. PMID- 10678242 TI - Methadone maintenance treatment: outcomes from the Otago methadone programme. AB - AIM: To provide information on methadone treatment outcomes for opiate-dependent individuals. METHODS: Questionnaires and random urine tests were completed for 112 Otago clients comparing outcomes before and during methadone maintenance treatment. RESULTS: Treatment retention rates were high, with 86% of clients remaining on the programme six months or more. The number of clients on benefits reduced by almost 30% during treatment, with employment rates doubling from 19% to 40% (including attendance at educational programmes). For the 89 clients injecting opiates daily at initial presentation, 64% reported no opiate use in the three months prior to review. Of the remaining 36%, opiate use reduced significantly. Rates of sharing injecting equipment reduced by almost 90%. Almost 50% of cannabis users reduced their use from daily to less than daily use. Clients reporting no current use of illicit benzodiazepines increased by 85%. Heavy binge drinking weekly or more reduced by almost 75%. Use of other illicit drugs reduced by almost 90%. Drug-related convictions reduced by almost 60%, while accidental drug overdoses reduced by over 90%. CONCLUSION: The widespread benefits of methadone maintenance treatment demonstrated underline the importance of making quality methadone programmes readily accessible within the health system. Currently, there are long waiting lists and many individuals cannot gain access to active treatment. We believe the health system urgently needs to look at expanding existing services and/or establishing private methadone clinics similar to those in New South Wales. PMID- 10678243 TI - Ophthalmomyiasis and nasal myiasis in New Zealand: a case series. AB - We report three cases of ophthalmomyiasis in New Zealand, due to the larvae of Oestrus ovis. All three patients reported eye injury caused by a fly. The larvae were removed from the conjunctival sac without difficulty under local anaesthesia. Presenting ocular symptoms of foreign body sensation, irritation, redness and photophobia all resolved swiftly. Topical antibiotic and steroid eye drops were administered. All three patients also developed nasal symptoms such as sneezing, nasal discharge and epistaxis. Otolaryngology follow-up demonstrated nasal myiasis in two patients which was treated with nasal decongestants. In addition, all three patients were treated with ivermectin (Mectizan). PMID- 10678244 TI - Cervical pregnancy managed without hysterectomy. PMID- 10678245 TI - Excessive use of inhaled salbutamol: the potential benefits of dose-reduction. A case report. AB - Inhaled, short-acting, beta-agonist medications are widely used in the treatment of asthma. It is recommended in current asthma management guidelines that these medications should only be used "as required" for relief of asthma symptoms. Despite this, there are still a number of asthmatic patients who are using excessive amounts. In occasional patients this may be detrimental to the control of asthma. We report the case of a 43-year-old, steroid-dependent asthmatic, with unstable asthma, using salbutamol up to 30 times a day. Following reduction of her beta-agonist use she had a dramatic improvement in her asthma control and was able to successfully stop her oral steroids. PMID- 10678246 TI - Breathlessness in older people. AB - Breathlessness in the elderly is a common clinical problem but should not be considered an inevitable consequence of the aging process. Because of the diverse causes and the possibility of more than one underlying mechanism, the approach to the breathless patient needs to be comprehensive with investigations guided by specific clinical questions. Once the underlying reversible factors have been identified and treated as far as practicable, management of the chronically breathless patient is based on symptom relief, exercise conditioning, optimisation of breathing patterns and patient education. Interventions should be objectively evaluated using symptom scores or a measure of exercise tolerance rather than a physiological measure alone. As there is the potential for harm (including cost), treatments offering no benefit should be promptly withdrawn. PMID- 10678247 TI - Immunisation coverage in New Zealand. PMID- 10678248 TI - Childhood immunisation in Rotorua. PMID- 10678249 TI - Third generation oral contraceptives and the risk of pulmonary embolism. PMID- 10678250 TI - Alternative therapies and "standard medicines". PMID- 10678251 TI - General practitioner found guilty of professional misconduct. PMID- 10678252 TI - Obesity research and the new century. PMID- 10678253 TI - Use of tape-recorded food records in assessing children's dietary intake. AB - BACKGROUND: Dietary assessment among children is particularly problematic when techniques are dependent on memory skills or advanced cognitive development. OBJECTIVE: The current study explored the use of self-report by tape recorders to document children's dietary intake immediately upon consumption, and compared this method with the traditional, interviewer-guided recall technique. In addition, the influence of body fatness and sociodemographic characteristics on the accuracy of recall and tape-recorded food records was determined. RESEARCH METHODS AND PROCEDURES: The sample included 30 black and white children aged 6.5 to 11.6 years (x = 9.5). Energy intake (EI), measured by six 24-hour food records (three for each method), was compared with total energy expenditure calculated by the doubly labeled water technique. Paired t tests, correlation analyses, and multiple regression analyses were performed. RESULTS: The analyses revealed poor validity of the tape recorder method (x misreporting score = -1.13+/-2.62 MJ/day, r for total energy expenditure and EI = -0.06, p = 0.74). Estimates of EI differed significantly between the tape recorder and recall methods (p<0.01). The traditional recall method was confirmed as a valid estimate of energy intake (x misreporting score = 0.04+/-2.38 MJ/day), although demonstrating a modest correlation with TEE (r = 0.32, p = 0.08). Although no significant predictors of misreporting using the recall method were identified in the multivariate analyses, older children and children with higher adiposity were more likely to misreport using the tape recorder method. DISCUSSION: The results suggest that the use of the tape recorder for estimating EI does not result in accurate assessments among children, although this technique may be useful for specific subgroups (i.e., younger and leaner children). PMID- 10678254 TI - Heart rate variability in obese children: relations to total body and visceral adiposity, and changes with physical training and detraining. AB - OBJECTIVE: Heart rate variability provides non-invasive information about cardiac parasympathetic activity (PSA). We determined in obese children: (1) relations of baseline PSA to body composition and hemodynamics; (2) effects of physical training (PT) and cessation of PT; and (3) which factors explained individual differences in responsivity of PSA to the PT. RESEARCH METHODS AND PROCEDURES: The root mean square of successive differences (RMSSD) was the index of PSA. Obese children (n = 79) were randomly assigned to groups that participated in PT during the first or second 4-month periods of the study. RESULTS: Baseline RMSSD was significantly (p<0.05) associated with lower levels of: fat mass, fat-free mass, subcutaneous abdominal adipose tissue, resting heart rate (HR), resting systolic blood pressure, and exercise HR. Stepwise multiple regression produced a final model (R2 = 0.36) that included only resting HR. The analysis of changes over the three time points of the study found a significant (p = 0.026) time by group interaction, such that RMSSD increased during periods of PT and decreased following cessation of PT. Greater individual increases in response to the PT (p<0.05) were seen in those who had lower pre-PT RMSSD levels, showed the greatest decreases in resting HR, and increased most in vigorous physical activity. The final regression model retained only the change in resting HR as a significant predictor of the changes in the RMSSD (R2 = 0.23). DISCUSSION: Regular exercise that improved fitness and body composition had a favorable effect on PSA in obese children. PMID- 10678255 TI - Abnormal signal-averaged electrocardiogram (SAECG) in obesity. AB - OBJECTIVE: The occurrence of small high-frequency electrocardiogram (ECG) potentials (1 to 20 microV) seen at the end of the QRS complex and into the ST segment have been correlated with increased risk for ventricular arrhythmias and sudden cardiac death. Computer-assisted analysis of these "late potentials" by signal-averaged electrocardiography (SAECG) has been studied and utilized to predict the likelihood of ventricular arrhythmias in various clinical states. Obesity is associated with significant cardiovascular morbidity and sudden death. Ventricular arrhythmias are postulated causes. We studied the occurrence of late potentials in a randomly selected group of obese patients and healthy volunteers. RESEARCH METHODS AND PROCEDURES: We performed SAECG on 105 subjects. Of these, 62 were obese ambulatory patients with body mass index (BMI) of >30 kg/m2, whereas 43 were healthy asymptomatic volunteers with a BMI of <30 kg/m2. Patients with a history of clinical heart disease and pulmonary disease, electrolyte abnormalities, recent hospitalizations, or abnormal screening ECG or taking medications known to alter the QRS interval were excluded. At least 250 beats were analyzed with a noise level of <0.50 microV. Criteria of a late potential include QRS duration >114 ms, high-frequency low amplitude >38 ms, and root-mean square voltage <20 microV. Patients were divided into four subgroups based on BMI values. The prevalence of SAECG abnormalities in each BMI subgroup was studied. We utilized multiple logistic regression analysis to study the effect of obesity, hypertension, and diabetes mellitus on abnormal SAECG results. RESULTS: Compared to age- and sex-matched healthy volunteers with BMI of <30 kg/m2, obese patients with BMI of >30 kg/m2 had significantly more abnormalities on SAECG (4.6% vs. 55%). In the obese group, the prevalence and number of abnormalities increased with increase in BMI (35% in the BMI 31 to 40 kg/m2 subgroup, 86% in the BMI 41 to 50 kg/m2 subgroup, and 100% in patients with BMI of >50 kg/m2). Multiple logistic regression analysis shows that BMI is an independent predictor variable of abnormal SAECG results in obese patients (n = 62) with BMI of >30 kg/m2 as well as in all study subjects (n = 105). BMI also predicts abnormality of each abnormal SAECG criterion in both obese and all subjects. Hypertension was found to influence the QRS duration alone in obese and all subjects. DISCUSSION: Obesity is associated with increased occurrence of abnormal SAECG results. These abnormalities are found both in obese patients with and without hypertension and/or diabetes. Obesity is an independent predictor variable of abnormal SAECG results. A history of hypertension predicts abnormality of QRS duration only. PMID- 10678256 TI - Leptin responses to weight loss in postmenopausal women: relationship to sex hormone binding globulin and visceral obesity. AB - OBJECTIVE: Leptin concentrations increase with obesity and tend to decrease with weight loss. However, there is large variation in the response of serum leptin levels to decreases in body weight. This study examines which endocrine and body composition factors are related to changes in leptin concentrations following weight loss in obese, postmenopausal women. RESEARCH METHODS AND PROCEDURES: Body composition (DXA), visceral obesity (computed tomography), leptin, cortisol, insulin, and sex hormone-binding globulin (SHBG) concentrations were measured in 54 obese (body mass index [BMI] = 32.0+/-4.5 kg/m2; mean +/- SD), women (60+/-6 years) before and after a 6-month hypocaloric diet (250 to 350 kcal/day deficit). RESULTS: Body weight decreased by 5.8+/-3.4 kg (7.1%) and leptin levels decreased by 6.6+/-11.9 ng/mL (14.5%) after the 6-month treatment. Insulin levels decreased 10% (p< 0.05), but mean SHBG and cortisol levels did not change significantly. Relative changes in leptin with weight loss correlated positively with relative changes in body weight (r = 0.50, p<0.0001), fat mass (r = 0.38, p<0.01), subcutaneous fat area (r = 0.52, p<0.0001), and with baseline values of SHBG (r = 0.38, p<0.01) and baseline intra-abdominal fat area (r = -0.27, p<0.06). Stepwise multiple regression analysis showed that baseline SHBG levels (r2 = 0.24, p<0.01), relative changes in body weight (cumulative r2 = 0.40, p<0.05), and baseline intra-abdominal fat area (cumulative r2 = 0.48, p<0.05) were the only independent predictors of the relative change in leptin, accounting for 48% of the variance. DISCUSSION: These results suggest that obese, postmenopausal women with a lower initial SHBG and more visceral obesity have a greater decrease in leptin with weight loss, independent of the amount of weight lost. PMID- 10678257 TI - Usefulness of anthropometry and DXA in predicting intra-abdominal fat in obese men and women. AB - OBJECTIVE: To investigate the usefulness of anthropometry and DXA in predicting intra-abdominal fat (IAF) in obese men and women. RESEARCH METHODS AND PROCEDURES: Observational, cross sectional study of 22 women and 18 men with a body mass index of 30 or above. IAF from 20 cm above and 10 cm below the L4 to L5 intervertebral disc was measured by magnetic resonance imaging (MRI) as a reference method. Central abdominal fat was measured from the upper border of L2 to the lower border of L4 by DXA. Waist and hip circumferences were also measured. RESULTS: In obese women DXA, waist circumference and waist-hip ratio were equally well correlated with IAF (r = 0.74, 0.75, and 0.70, respectively). In obese men DXA was moderately correlated with IAF measured by MRI (r = 0.46), whereas waist circumference and waist-hip ratio were not significantly correlated with IAF. DISCUSSION: The prediction of IAF in obese subjects was highly dependent on sex more than in non-obese persons. Anthropometry and DXA were equally useful in obese women, whereas anthropometry had no predictive power and DXA was the only acceptable predictor of IAF in obese men. PMID- 10678258 TI - Diabetes disease stage predicts weight loss outcomes with long-term appetite suppressants. AB - OBJECTIVES: Characterize degree of weight loss with stage of diabetes and describe its effect on cardiovascular disease risk factors in obese patients with and without diabetes. RESEARCH METHODS AND PROCEDURES: Retrospective cohort analysis from patients participating in a long-term weight management protocol using diet, exercise, behavioral modification, and appetite-suppressant therapy. Patient groups, with (n = 19) and without diabetes (n = 19) were matched for age, gender, and weight before weight loss therapy. The effect of 12 months of therapy on weight, blood pressure, glycemic control, lipid profile, and medication requirements were tested. Additionally, patients were grouped or staged based upon therapy required for control of diabetes at the beginning of weight loss intervention. Analysis of covariance described relationships between diabetes disease stage and weight loss at 12 months. RESULTS: Nondiabetic patients had greater mean reduction in BMI than the diabetic group (7.98 kg/m2 vs. 4.77 kg/m2, p<0.01). A significant linear trend (p<0.001) for decreasing weight loss with stage of diabetes was observed. Blood pressure, lipid profile, and glycemia improved significantly. The average daily glyburide-equivalent dose decreased from 9.4 to 3.0 mg (p<0.01). DISCUSSION: Patients with diabetes lost less weight than similarly obese patients without diabetes. Regardless of differential weight loss between groups, cardiovascular disease risk factors improved. Hypoglycemic medication requirements decreased with weight loss therapy. A predictive relationship may exist between diabetes disease stage before weight loss therapy and future weight loss potential. PMID- 10678259 TI - Weight loss, weight maintenance, and improved cardiovascular risk factors after 2 years treatment with orlistat for obesity. European Orlistat Obesity Study Group. AB - OBJECTIVE: To determine the effect of orlistat, a new lipase inhibitor, on long term weight loss, to determine the extent to which orlistat treatment minimizes weight regain in a second year of treatment, and to assess the effects of orlistat on obesity-related risk factors. RESEARCH METHODS AND PROCEDURES: This was a 2-year, multicenter, randomized, double-blind, placebo-controlled study. Obese patients (body mass index 28 to 43 kg/m2) were randomized to placebo or orlistat (60 or 120 mg) three times a day, combined with a hypocaloric diet during the first year and a weight maintenance diet in the second year of treatment to prevent weight regain. Changes in body weight, lipid profile, glycemic control, blood pressure, quality of life, safety, and tolerability were measured. RESULTS: Orlistat-treated patients lost significantly more weight (p<0.001) than placebo-treated patients after Year 1 (6.6%, 8.6%, and 9.7% for the placebo, and orlistat 60 mg and 120 mg groups, respectively). During the second year, orlistat therapy produced less weight regain than placebo (p = 0.005 for orlistat 60 mg; p<0.001 for orlistat 120 mg). Several obesity-related risk factors improved significantly more with orlistat treatment than with placebo. Orlistat was generally well tolerated and only 6% of orlistat-treated patients withdrew because of adverse events. Orlistat leads to predictable gastrointestinal effects related to its mode of action, which were generally mild, transient, and self-limiting and usually occurred early during treatment. DISCUSSION: Orlistat administered for 2 years promotes weight loss and minimizes weight regain. Additionally, orlistat therapy improves lipid profile, blood pressure, and quality of life. PMID- 10678260 TI - Lipoprotein subpopulation distributions in lean, obese, and type 2 diabetic women: a comparison of African and white Americans. AB - OBJECTIVE: Abnormal subpopulation distributions of plasma lipoproteins have been reported in white American (WA) women with obesity and type 2 diabetes that explain part of the elevated rate of cardiovascular disease in these patients. This study examined if these perturbations also occur in obese and diabetic African American (AA) women and compared the lipoprotein profiles with WA counterparts. RESEARCH METHODS AND PROCEDURES: We determined the lipoprotein subpopulation distribution in the plasma of 51 lean women (29 WA, 22 AA, body mass index [BMI] < 30), 50 obese women (27 WA, 23 AA, BMI > 30), and 43 obese women with type 2 diabetes (27 WA, 16 AA), by nuclear magnetic resonance spectroscopy. RESULTS: AA diabetic women, like WA diabetic women, had a larger average very low density lipoprotein (VLDL) size, elevated levels of small low density lipoprotein cholesterol (LDL-C), and lower levels of small high density lipoprotein cholesterol (HDL-C), when compared to lean controls (p<0.05). These differences were accompanied by higher VLDL-triglycerides (TG) and LDL-C in WA (p<0.05), but not in AA. Although the effects of obesity and diabetes on lipoprotein subpopulation were fairly similar for AA and WA, some racial differences, particularly with respect to HDL, were observed. DISCUSSION: The atherogenic perturbations in lipoprotein profiles of obese AA women, particularly those with diabetes, were relatively similar to those found in WA women and may be contributing to the increased rate of cardiovascular disease (CVD) in AA with obesity and diabetes. The parameters of subpopulation distribution may provide better markers for CVD than lipid concentrations alone, particularly in AA women. Furthermore, subtle racial differences in lipoprotein profiles suggest that race specific criteria may be needed to screen patients for CVD. PMID- 10678261 TI - Six-month treatment of obesity with sibutramine 15 mg; a double-blind, placebo controlled monocenter clinical trial in a Hispanic population. AB - OBJECTIVE: To evaluate the safety and efficacy of sibutramine 15 mg by mouth once per day in obese patients over a period of 6 months. RESEARCH METHODS AND PROCEDURES: A monocenter, double-blind, placebo controlled, parallel, prospective clinical trial was carried out. Sixty-nine male and female obese patients (body mass index [BMI] > 30 kg/m2) aged 16 to 65 years entered the trial. RESULTS: 22 of 35 patients in the sibutramine group and 9 of 34 patients in the placebo group completed the trial. The high dropout rate in the sibutramine group was due to adverse events in 3 cases, lack of efficacy (as judged by patients) in 7, loss to follow-up in 2, and an orthopedic device being worn in 1; in the placebo group the dropouts were ascribed to lack of efficacy (as judged by patients) in 17 cases and to loss to follow-up in 8 cases. Using the method of last observation carried forward, the weight loss in the sibutramine group was 10.27 kg (95% confidence intervals [95% CI] 7.66; 13.07) and 1.26 kg (95% CI 0.3; 2.23) in the placebo group. The BMI loss was 4.17 kg/m2 (95% CI 3.11; 5.22) in the sibutramine group and 0.53 kg/m2 (95% CI 0.13; 0.92) in the placebo group. The waist circumference reduction was 12.51 cm (95% CI 9.25; 15.77) in the sibutramine group and 3.26 cm (95% CI 1.38; 5.14) in the control group (p<0.05 by paired Student's t test for all the intragroup comparisons). Twenty-three sibutramine patients had 34 adverse events, the most frequent adverse events in the sibutramine group were upper respiratory tract infections (n = 6) and constipation (n = 6); 16 placebo patients had 21 adverse events. Three sibutramine patients withdrew their informed consent when they had adverse events. DISCUSSION: The results show that sibutramine induces significant loss of body weight and waist circumference. Cardiovascular function was not significantly affected by sibutramine. Sibutramine was well tolerated by most of the patients. PMID- 10678262 TI - The expression of peroxisome proliferator-activated receptor gamma in pig fetal tissue and primary stromal-vascular cultures. AB - OBJECTIVE: This study was designed to determine when peroxisome proliferator activated receptor gamma (PPARgamma) is expressed in developing fetal adipose tissue and stromal-vascular adipose precursor cells derived from adipose tissue. In addition we examined developing tissue for CCAAT/enhancer-binding protein beta (C/EBPbeta) expression to see if it was correlated with PPARgamma expression. Pituitary function and hormones involved with differentiation (dexamethasone and retinoic acid) were also tested for their effects on PPARgamma expression to determine if hormones known to affect differentiation also effect PPARgamma expression in vivo and in cell culture. RESEARCH METHODS AND PROCEDURES: Developing subcutaneous adipose tissues from the dorsal region of the fetal pig were collected at different gestation times and assayed using Western blot analysis to determine levels of PPARgamma and C/EBPbeta. Hypophysectomy was performed on 75-day pig fetuses and tissue samples were then taken at 105 days for Western blot analysis. Adipose tissue was also taken from postnatal pigs to isolate stromal-vascular (S-V) cells. These adipose precursor cells were grown in culture and samples were taken for Western blot analysis to determine expression levels of PPARgamma. RESULTS: Our results indicate that PPARgamma is expressed as early as 50 days of fetal development in adipose tissue and continues through 105 days. Expression of PPARgamma was found to be significantly enhanced in adipose tissue from hypophysectomized fetuses at 105 days of fetal development (p<0.05). C/EBPbeta was not found in 50- or 75-day fetal tissues and was found only at low levels in 105-day tissues. C/EBPbeta was not found in hypophysectomized (hypoxed) 105-day tissue where PPARgamma was elevated. S-V cells freshly isolated from adipose tissue of 5- to 7-day postnatal pigs showed the expression of PPARgamma1. When S-V cells were cultured, both PPARgamma1 and 2 were expressed after the first day and continued as cells differentiated. High concentrations of retinoic acid decreased PPARgamma expression in early S-V cultures (p<0.05). DISCUSSION: Our data indicate that PPARgamma is expressed in fetal adipose tissue very early before distinct fat cells are observed and can be expressed without the expression of C/EBPbeta. The increase in PPARgamma expression after hypophysectomy may explain the increase in fat cell size under these conditions. Adipose precursor cells (S-V cells) from 5- to 7-day postnatal pigs also express PPARgamma in the tissue before being induced to differentiate in culture. Thus S V cells from newborn pig adipose tissue are probably more advanced in development than the 3T3-L1 cell model. S-V cells may be in a state where PPARgamma and C/EBPalpha are expressed but new signals or vascularization are needed before cells are fully committed and lipid filling begins. PMID- 10678263 TI - The human obesity gene map: the 1999 update. AB - This report constitutes the sixth update of the human obesity gene map incorporating published results up to the end of October 1999. Evidence from the rodent and human obesity cases caused by single gene mutations, Mendelian disorders exhibiting obesity as a clinical feature, quantitative trait loci (QTL) uncovered in human genome-wide scans and in crossbreeding experiments with mouse, rat, pig and chicken models, association and linkage studies with candidate genes and other markers is reviewed. Twenty-five human cases of obesity can now be explained by variation in five genes. Twenty Mendelian disorders exhibiting obesity as one of their clinical manifestations have now been mapped. The number of different QTLs reported from animal models reaches now 98. Attempts to relate DNA sequence variation in specific genes to obesity phenotypes continue to grow, with 89 reports of positive associations pertaining to 40 candidate genes. Finally, 44 loci have linked to obesity indicators in genomic scans and other linkage study designs. The obesity gene map depicted in Figure 1 reveals that putative loci affecting obesity-related phenotypes can be found on all autosomes, with chromosomes 14 and 21 showing each one locus only. The number of genes, markers, and chromosomal regions that have been associated or linked with human obesity phenotypes continues to increase and is now well above 200. PMID- 10678264 TI - Population advice on salt restriction: the social issues. AB - The scientific evidence that underlies public health advice depends upon critical integration of information from several sources. The most informative evidence relating to the effects of population reduction in salt intake comes from systematic reviews of clinical trials. Recent rigorous reviews of salt restriction trials in normal subjects show extremely small effects ranging from 1 to 2 mm Hg for systolic blood pressure and 0.1 to 1.0 mm Hg for diastolic pressure. These are the result of much greater reductions in sodium intake than can be achieved by population advice, and may be further amplified by publication bias and effects of changes in other dietary components. There is little trial evidence to enable possible benefits and adverse effects to be balanced. Reviews biased by the inclusion of nonrandomized studies exaggerate the apparent blood pressure fall 5- to 50-fold and create spurious apparent progressive falls in blood pressure. Nevertheless, citation analysis shows that they are quoted much more frequently than rigorous reviews reaching more negative conclusions. This appears to be the result of an attempt to create an impression of scientific consensus. The salt debate has important implications for social policy. PMID- 10678265 TI - Evidence in favor of moderate dietary sodium reduction. PMID- 10678266 TI - Plasma homocysteine concentrations and insulin sensitivity in hypertensive subjects. AB - Hyperhomocysteinemia is associated with several cardiovascular disease risk factors including endothelial dysfunction and abnormalities of clotting functions, which are also common features of insulin resistance syndrome observed in hypertensive patients. Recent study has shown that acute hyperinsulinemia can lower plasma homocysteine concentrations in nondiabetic but not in type 2 diabetic individuals, indicating that insulin may regulate homocysteine metabolism. To investigate the relationships between plasma homocysteine concentration and insulin sensitivity, we studied 90 Chinese hypertensive patients and a group of control subjects (n = 86) matched for age, gender, and body mass index. Fasting plasma homocysteine levels, plasma lipoprotein concentrations, plasma glucose, and insulin responses to oral glucose tolerance tests (OGTT) were determined. The results showed that fasting plasma homocysteine concentrations were significantly higher in subjects with hypertension than in those with normotension (mean +/- SEM, 8.1 +/- 0.6 v 6.8 +/- 0.2 micromol/L; P < .05). Fasting plasma homocysteine levels correlated significantly with insulin secretion in response to OGTT even after adjustment for body mass index (P < .05) in hypertensive patients but not in normotensive individuals. However, fasting plasma homocysteine concentrations showed no correlations with steady-state plasma glucose concentration, a measurement of insulin sensitivity, during an insulin suppression test in groups of hypertensive (n = 42) and normotensive (n = 37) subjects. When the steady-state plasma glucose concentrations were divided into three tertiles, fasting plasma homocysteine concentrations showed no difference across these three groups in either hypertensive patients (8.6 +/- 0.5 v 7.2 +/- 0.5 v 8.4 +/- 0.6 micromol/L; P = .148) or normotensive subjects (6.3 +/- 0.4 v 8.0 +/- 0.8 v 7.0 +/- 0.8 micromol/L; P = .199). In conclusion, hypertensive Chinese subjects had higher fasting plasma homocysteine concentrations and a higher degree of insulin resistance when compared to a group of age-, gender-, and body mass index-matched normotensive individuals. Fasting plasma homocysteine levels were associated with insulin response to OGTT in hypertensives but not in normotensives. No correlation was observed between the degree of insulin resistance and plasma homocysteine levels in either the hypertensive or the normotensive group. The role of insulin in homocysteine metabolism deserves further investigation. PMID- 10678267 TI - Effects of hyperinsulinemia on sympathetic responses to mental stress. AB - In a recent study, we could not find evidence to support the hypothesis that insulin activates the sympathetic nervous system (SNS) during a hyperinsulinemic glucose clamp procedure. Mental stress tests (MST), however, may be used to detect differences in blood pressure and SNS activity that are not present during baseline or resting conditions. In this study, we aimed to investigate the effects of hyperinsulinemia during glucose clamp on blood pressure and sympathetic responses to mental stress. Borderline hypertensive but otherwise healthy 21-year-old men (n = 18) underwent 5 min of mental arithmetic stress testing (MST-1) before and at the end of 120 min of isoglycemic hyperinsulinemic glucose clamp (MST-2) with infusion rates of glucose and insulin kept constant. Insulin concentration increased from 119 +/- 10 pmol/L to 752 +/- 65 pmol/L. We observed highly significant increases in blood pressure and heart rate in response to MST, but neither insulin nor saline solution infusions affected these responses. During MST-1, norepinephrine increased by 461 +/-165 pmol/L (mean +/- SEM) and epinephrine by 218 +/- 76 pmol/L. During MST-2 the changes were 372 +/- 112 pmol/L and 187 +/- 60 pmol/L, respectively. The norepinephrine (P = .8) and epinephrine (P = .7) responses were unchanged by insulin. Thus, there were similar increases in blood pressure, heart rate, and plasma catecholamine concentrations in arterialized venous blood in response to MST despite the infusion of insulin. A possible time effect was excluded by including a saline solution control group (n = 7) that showed almost identical results. Our results suggest that acute hyperinsulinemia during isoglycemic glucose clamp does not interfere with cardiovascular or sympathetic responses to mental stress. PMID- 10678268 TI - Sympathoexcitatory responses to the acute blood pressure fall induced by central or peripheral antihypertensive drugs. AB - This study was designed to evaluate the effects of an acute blood pressure reduction brought about by a peripheral vasodilator agent (prazosin) or by a drug combining central and peripheral modes of action (urapidil), on three markers of adrenergic tone such as muscle sympathetic nerve traffic (MSNA), venous plasma norepinephrine (NE), and heart rate (HR). In 12 untreated essential hypertensives (age, 50.7 +/- 1.9 years; mean +/- SEM), we evaluated in two experimental sessions, according to a double-blind crossover design, the effects of acute oral administration of 2 mg prazosin or 30 mg urapidil on beat-to-beat finger blood pressure (Finapres), HR (electrocardiogram), NE (high-performance liquid chromatography), and MSNA (microneurography at a peroneal nerve). In each session measurements were performed in the no-drug control state and repeated throughout a 3-h period after drug administration. For similar blood pressure reductions, the two drugs caused similar increases in NE and MSNA (peak effects: NE = +1.1 +/ 0.2 vs 0.9 +/- 0.2 nmol/L and MSNA = +10.9 +/- 1.8 vs +10.1 +/- 1.6 bursts/min for prazosin and urapidil respectively, P = ns between drugs), whereas HR increased more markedly after prazosin administration (+6.1 +/- 1.1 vs +2.4 +/- 0.8 beats/min, P < 0.05). These data provide evidence that acute blood pressure reductions induced by antihypertensive drugs with central or peripheral modes of action activate the sympathetic nervous system to a similar extent. Thus adrenergic activation is not peculiar to vasodilators but rather generalized to any drug-induced acute blood pressure fall, presumably because of the lack of a baroreflex resetting, which occurs during chronic but not during acute antihypertensive treatment. PMID- 10678269 TI - Changes in clinical features and long-term prognosis in patients with pheochromocytoma. AB - To investigate changes in preoperative clinical features and the long-term outcome of tumor recurrence, mortality, and morbidity in patients with pheochromocytoma, we retrospectively examined changes in the clinical features by comparing 49 patients from 1957 to 1985 (group I) with 46 patients from 1986 to December 1995 (group II). In addition in these 95 patients (excluding 2 who had died before operation), we evaluated long-term postoperative outcome from the initial operation to August 1996 (909 patient-years). The mean age in group II was older than that of group I. The percentage of patients having proteinuria or hypertensive retinopathy in group II was less than that in group I. Of 20 patients with incidentally discovered pheochromocytoma, 7 (35%) were > or =60 years old, 7 asymptomatic, and 11 (55%) normotensive. Plasma and urinary catecholamines in these patients were significantly (P < .01) lower than in patients with pheochromocytoma having typical clinical features. Long-term cohort study showed 14 deaths. Relative survival rates were 91% at 5 years and 83% at 10 years and unchanged thereafter. The Kaplan-Meier estimate of pheochromocytoma free survival was shorter in patients with a larger-than-median (60 g) tumor weight. Six patients had malignant recurrence 3 to 101 months (median, 45 months) after the initial operation. Of 65 patients confirmed alive at follow-up, 11 were hypertensive. In the Cox model, hypertension-free survival was not associated with age, a family history of hypertension, duration of hypertension, or creatinine clearance. Pheochromocytoma should be diagnosed from a wide spectrum of clinical features including those that are not generally suspected of resulting from excess catecholamines or hypertension, and after surgery, patients with this disease should be followed-up carefully for a long period (at least 10 years) because of the risk of tumor recurrence and the high prevalence of disease. PMID- 10678270 TI - Ambulatory blood pressure after acute exercise in older men with essential hypertension. AB - We sought to determine whether reductions in blood pressure in hypertensives after acute exercise persist for more than the 2 to 3 h found in controlled laboratory settings. Subjects (n = 11) were obese (32 +/- 4% body fat), sedentary (VO2max 27 +/- 4 mL/kg/min) 60 +/- 6-year-old men with stage 1 or 2 essential hypertension. Ambulatory blood pressure was recorded on 1 day preceded by 45 min of 70% VO2max treadmill exercise and on another day not preceded by exercise. Systolic blood pressure was lower by 6 to 13 mm Hg for the first 16 h after exercise (P < .05) compared to the day without prior exercise. Twenty-four-hour, day, and night average systolic blood pressures were significantly lower on the day after exercise. There was a trend for peak systolic blood pressure to be lower during the entire 24 h and the day portion of the recording; peak systolic blood pressure was significantly lower during the night portion of the recording after exercise. Systolic blood pressure load (percent of systolic blood pressure readings >140 mm Hg) was reduced during the entire 24 h and the day portion of the recording after exercise. Diastolic blood pressure was lower for 12 of the first 16 h after acute exercise (hours 0 to 4, 5 to 8, 13 to 16) (P < .05) compared to the day without prior exercise. Twenty-four-hour, day, and night average diastolic blood pressure was also significantly lower on the recording after exercise. Peak diastolic blood pressure was lower over the entire 24-h period. Diastolic blood pressure load (percent of diastolic blood pressure readings >90 mm Hg) was lower during the entire 24 h and the day portion of the day after exercise. Preliminary data also suggest that common genetic polymorphisms at the angiotensinogen, lipoprotein lipase, and angiotensin converting enzyme loci may affect the blood pressure-lowering response after acute exercise. Thus, in sedentary, obese hypertensive men a single aerobic exercise session reduced blood pressure enough to result in significantly lower 24-h average systolic, diastolic, and mean arterial blood pressure. This could result in a reduced cardiovascular load during the 24 h after acute exercise in older hypertensive men. PMID- 10678271 TI - Predictors of interindividual variation in ambulatory blood pressure and their time or activity dependence. AB - The objectives of this study were to determine whether total interindividual variation in blood pressure (BP) differs between inactive and active hours of the day, to identify predictors of interindividual variation in BP, and to assess whether variation associated with any of these identified predictors is greater (or less) during inactive hours than during active hours of the day. We obtained ambulatory BP recordings over 20 consecutive hours (12 active, out of bed [daytime]; and 8 inactive, in bed [nighttime]) in a sample of 240 unrelated, non Hispanic white adults (138 men; 102 women). We estimated total interindividual variation in BP, and the percentage of interindividual variation associated with measures of age and body size, metabolic traits, catecholamines, erythrocyte cation transport, and renal function. We used linear regression to assess changes in the hourly estimates of total interindividual variation and in variation attributable to each set of predictor traits over the 20 h. In both men and women, total interindividual variation in systolic BP was significantly greater (not less) during inactive hours than during active hours. In addition, in women, total interindividual variation in diastolic BP was as great during inactive hours as during active hours. Each set of traits considered predicted a statistically significant percentage of interindividual variation in BP. None of the sets of traits predicted a greater percentage of interindividual variation during the inactive hours than during the active hours. Measures of age and body size, catecholamines, cation transport and renal function traits predicted significantly less interindividual variation during inactive hours than during active hours of the day. That total interindividual variation in BP is as great or greater during inactive hours than during active hours of the day emphasizes the potential for differences in nighttime BP to contribute to the development of cardiovascular disease. In as much as the predictors of interindividual variation in BP differ between the daytime and nighttime, the causes of variation during these two times may also differ. PMID- 10678272 TI - Resistance to activated protein C and FV leiden mutation in patients with a history of acute myocardial infarction or primary hypertension. AB - This study was designed to investigate both resistance to activated protein C (APC-R) and the factor FV Q506 mutation incidence in patients with a history of acute myocardial infarction (AMI) and patients with primary hypertension (PH), a high-risk group for arterial thrombosis. Eighty patients with a history of AMI (group A), 160 patients with a history of PH (group B), and 124 age-matched controls without arterial disease (group C) were studied. APC-R was determined using the Coatest APC Resistance Kit of Chromagenix, Sweden. The prevalence of the FV Q506 mutation was estimated by DNA analysis (Bertina method). The prevalence of the FV Q506 mutation was 20%, 13.75%, and 8% in groups A, B, and C, respectively (A v C P = .0466). The prevalence of APC-R was 47.5% in group A v 13% in group C (P < .0001) and 36.25% in group B v 13% in group C (P < .0001). The response to activated protein C expressed as mean value +/- SD was 2.05 +/- 0.33 in group A v 2.56 +/- 0.46 in group C (P < .05) and 2 +/- 0.22 in group B v 2.56 +/- 0.46 in group C (P < .05). These findings suggest that patients with a history of AMI or PH have a significantly increased incidence of both APC-R and FV Q506 mutation compared with the control group. These findings support the hypothesis that these anticoagulant defects may be risk factors for arterial thrombosis. PMID- 10678273 TI - Human chorionic gonadotrophin is an endothelium-independent inhibitor of rat aortic smooth muscle contractility. AB - The study tested the hypothesis that human chorionic gonadotrophin (hCG) attenuates isolated vascular smooth muscle contractility and investigated the role of the vascular endothelium in hCG-induced altered responses of vascular smooth muscle. The contractile responses of isolated aortic rings from normal, hCG-treated, and estrogen-treated female virgin Wistar rats to phenylephrine, angiotensin II, KCl, and CaCl2 were compared. The effect of pretreatment with N monomethyl-L-arginine (L-NMMA), methylene blue, indomethacin, calcium-free medium, and de-endothelialization on responses to phenylephrine of aortic rings from control and hCG-treated rats were also examined. Intraperitoneal administration of hCG caused attenuation of contractile responses of isolated aortic rings to all agents. The attenuated responses to phenylephrine were not reversed by de-endothelialization, or pretreatment of the rings with L-NMMA, methylene blue, or indomethacin. It was concluded that hCG attenuates vascular smooth muscle contractility. The effect is independent of the vascular endothelium, not agonist-specific, and appears to involve alterations of calcium availability. PMID- 10678274 TI - Peroxisome proliferator-activated receptor (PPAR)-gamma expression in human vascular smooth muscle cells: inhibition of growth, migration, and c-fos expression by the peroxisome proliferator-activated receptor (PPAR)-gamma activator troglitazone. AB - This study was conducted to determine whether cultured human coronary artery and aorta vascular smooth muscle (VSM) cells express the nuclear transcription factor peroxisome proliferator-activated receptor-gamma (PPARgamma); whether the thiazolidinedione troglitazone, a ligand for PPARgamma, would inhibit c-fos expression by these cells; and whether troglitazone would inhibit proliferation and migration induced in these cells by mitogenic growth factors. Using immunoblotting and reverse-transcriptase polymerase chain reaction (RT-PCR) techniques, we show that both human aorta and coronary artery VSM cell lines expressed PPARgamma protein and mRNA for both PPARgamma isoforms, PPARgamma1 and PPARgamma2. Immunocytochemical staining localized the PPARgamma protein primarily within the nucleus. Troglitazone inhibited basic fibroblast growth factor and platelet-derived growth factor-BB induced DNA synthesis in a dose-dependent manner and downregulated the growth-factor-induced expression of c-fos. Troglitazone also inhibited the migration of coronary artery VSM cells along a platelet-derived growth factor-BB concentration gradient. These findings demonstrate for the first time the expression and nuclear localization of PPARgamma in human coronary artery and aorta VSM cells. The data also suggest that the downregulation of c-fos expression, growth-factor-induced proliferation, and migration by VSM may, in part, be mediated by activation of the PPARgamma receptor. PMID- 10678275 TI - Modulation of endothelin-1 coronary vasoconstriction in spontaneously hypertensive rats by the nitric oxide system. AB - To determine whether nitric oxide contributes to the augmented vasoconstrictive response to endothelin-1 (ET-1) in coronary vessels of hypertensive hearts, and also whether L-arginine administration can inhibit the augmented response to ET 1, we designed experiments to measure coronary perfusion resistance in isolated hearts of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) with or without L-arginine administration (0.5 g/L) for 2 weeks. The hearts were paced at a constant rate and perfused by the Langendorff technique at constant pressure (75 mm Hg). Perfusion flow and pressure were monitored, and coronary vascular resistance (CVR) was calculated. ET-1 infusion elicited dose dependent increases in CVR in both WKY and SHR. At an ET-1 concentration of 1.5 x 10(-9) mol/L, the response was significantly greater in SHR. In L-NAME-treated WKY and SHR, responses to ET-1 were augmented, compared with those of nontreated rats, and this augmentation was greater in WKY. L-arginine administration reduced the CVR response to ET-1 in SHR, whereas it did not change responses to ET-1 in WKY. These findings suggest that the augmented vasoconstriction of the coronary artery induced by ET-1 in hypertensive hearts was due to a reduction in nitric oxide release in coronary vessels and that L-arginine can partially inhibit the vasoconstrictive response of the coronary artery. PMID- 10678276 TI - Influence of correction for within-person variability in blood pressure on the prevalence, awareness, treatment, and control of hypertension. AB - We assessed the influence of correction for within-person variability (WPV) on the prevalence, awareness, treatment, and control of hypertension. Data were collected from two cross-sectional population-based studies on cardiovascular disease risk factors from 1987 to 1995 among 56,026 subjects aged 20 to 59 years. Correction factors were calculated from an internal reproducibility study among 924 subjects who were examined in 1989 and 1990. The prevalence of hypertension without a correction of blood pressure values for WPV was substantially overestimated (38%), whereas the prevalence of awareness and treatment of hypertension were substantially underestimated (-13% and -28%). The prevalence of control of hypertension did not change much after this correction. It may be advisable to perform a correction for within-person variability to obtain valid prevalence estimates in surveys that only take one or two measurements of blood pressure. PMID- 10678277 TI - Reproducibility of heart rate measured in the clinic and with 24-hour intermittent recorders. AB - This study was undertaken to assess the reproducibility of office versus ambulatory heart rates in 839 hypertensive subjects participating in the Hypertension and Ambulatory Recording Venetia Study (HARVEST). A 24-hour heart rate was recorded twice; this procedure was repeated three months later. Reproducibility was better for ambulatory than for office measurement, and was greater for 24-hour than for daytime heart rate, and lowest for night-time heart rate. Reproducibility of office heart rate was impaired above 85 bpm, and was poorer in subjects with more severe office hypertension. A small but significant decrease in average daytime (-1 bpm, P < 0.0001) and virtually no change in night time heart rate (-0.3 bpm, NS) were observed at repeat recording. Heart rate reproducibility indices were related to the extent of the heart rate and blood pressure white-coat effect, but did not vary according to age, gender, body mass index, day-night blood pressure difference, or alcohol or tobacco use. Results indicate that heart rate recorded over the 24 hours has a better reproducibility than office heart rate, and could thus be a better prognostic indicator than traditional measurement of resting heart rate in the hospital setting. PMID- 10678278 TI - Chronic intravenous glucose infusion causes moderate hypertension in rats. AB - We have reported that chronic insulin infusion increases mean arterial pressure (MAP) in rats. In those studies, glucose was coinfused to prevent hypoglycemia, but it is possible that the glucose infusion rate may have exceeded the rate actually required to prevent hypoglycemia. If true, then the glucose infusion alone should have a similar effect, and this study tested that hypothesis. In six rats (insulin group) instrumented with artery and vein catheters, insulin was infused for 7 days intravenously (iv) at 1.5 mU/kg/min together with glucose iv at 18.6 mg/kg/min. Seven other rats (glucose group) received the same glucose infusion for 7 days but without iv insulin. MAP increased significantly in both groups, from 98 +/- 3 and 96 +/- 2 mm Hg to 107 +/- 5 and 104 +/- 3 mm Hg in the insulin and glucose groups, respectively, and the renal and hormonal changes were similar to those previously reported during insulin infusion. There were no significant differences between the two groups for any variable measured. These data indicate that the sugar intake provided by the glucose infusion essentially mimics the response to our insulin and glucose infusion protocol, and that similar mechanisms underlie the renal and cardiovascular responses to each protocol. PMID- 10678279 TI - Statement from the National High Blood Pressure Education Program: prevalence of hypertension. PMID- 10678280 TI - What level of plasma homocyst(e)ine should be treated? Effects of vitamin therapy on progression of carotid atherosclerosis in patients with homocyst(e)ine levels above and below 14 micromol/L. AB - High levels of plasma homocyst(e)ine (H[e]) are associated with increased vascular risk. Treatment is being contemplated, but the level at which patients should be treated is not known. We compared the response of carotid plaque to vitamin therapy in patients with H(e) above and below 14 micromol/L, a level commonly regarded as high enough to warrant treatment. Two-dimensional B-mode ultrasound measurement of carotid plaque was used to assess the response to vitamin therapy with folic acid 2.5 mg, pyridoxine 25 mg, and cyanocobalamin 250 microg daily, in 101 patients with vascular disease (51 with initial plasma levels above, and 50 below 14 micromol/L). Among patients with plasma H(e) >14 micromol/L, the rate of progression of plaque area was 0.21 +/- 0.41 cm2/year before vitamin therapy, and -0.049 +/- 0.24 cm2/year after vitamin therapy (P2 = .0001; paired t test). Among patients with levels <14 micromol/L, the rate of progression of plaque was 0.13 +/- 0.24 cm2/year before vitamin therapy and 0.024 +/- 0.29 cm2/year after vitamin therapy (P2 = .022, paired t test). The change in rate of progression was -0.15 +/- .44 cm2/year below 14 micromol/L, and -0.265 +/- 0.46 cm2/year above 14 micromol/L (P = 0.20). Vitamin therapy regresses carotid plaque in patients with H(e) levels both above and below 14 micromol/L. These observations support a causal relationship between homocyst(e)ine and atherosclerosis and, taken with epidemiologic evidence, suggest that in patients with vascular disease, the level to treat may be <9 micromol/L. PMID- 10678281 TI - Carotid thickening, cardiac hypertrophy, and angiotensin converting enzyme gene polymorphism in patients with hypertension. AB - An insertion/deletion (I/D) polymorphism of the angiotensin converting enzyme (ACE) gene has been associated with increased risk for myocardial infarction, cardiomyopathy, carotid thickening, and cardiac hypertrophy. However, a conclusive agreement about the role of ACE genotype in the genetics of cardiovascular disease has not yet been reached. This study was undertaken to investigate the relationship of the I/D polymorphism of the ACE gene with carotid intima-media thickness (IMT) and left ventricular mass (LVM) in 175 Chinese patients with mild-to-moderate hypertension. The I/D genotypes were detected by the polymerase chain reaction using primers flanking the polymorphic region in intron 16 of the ACE gene. The IMT was measured in the common carotid and carotid bifurcation by B-mode ultrasound. The LVM was calculated with M-mode echocardiographic measures of the left ventricle. Patients with the DD genotype (n = 41) showed significant greater carotid IMT (1.593 +/- 0.879 v 1.309 +/- 0.703 and 1.171 +/- 0.583 mm, P = .01) but insignificant higher LVM index (123.8 +/- 36.6 v 123.7 +/- 37.4 and 118.2 +/- 33.0 g/m2, P = .61) than did those with the DI (n = 69) and II (n = 65) genotypes. The deletion polymorphism of the ACE gene (P = .04) was a significant predictor for carotid IMT on multiple regression analysis, controlling all the potential confounding factors including age (P = .001), systolic blood pressure (P = .09), smoking (P = .08), and plasma tissue plasminogen activator antigen (P = .03), but the LVM correlated only with age (P = .02), sex (P < .001), and body mass index (P < .001). These results indicated that the DD genotype of the ACE gene could be considered a risk factor for the development of early atherosclerosis in carotid arteries but not for left ventricular hypertrophy in the hypertensive population. PMID- 10678282 TI - Risk stratification in hypertension: new insights from the Framingham Study. AB - Five decades of epidemiologic research have established that blood pressure elevation is a common and powerful contributor to all of the major cardiovascular diseases, including coronary disease, stroke, peripheral artery disease, renal disease, and heart failure. The common variety of hypertension designated benign essential hypertension was not shown to be either benign or essential. Although clinicians favor the diagnosis and treatment of hypertension in terms of diastolic blood pressure elevation and categoric cut points, epidemiologic data show a more important influence of systolic blood pressure, and a continuous, graded influence of blood pressure even within what is regarded as the normotensive range. An important revelation in epidemiologic hypertension research is that hypertension usually occurs in conjunction with other metabolically linked risk factors; therefore, less than 20% occurs in isolation. The other risk factors that tend to accompany hypertension include glucose intolerance, obesity, left ventricular hypertrophy, and dislipidemia (elevated total, LDL, and small dense LDL cholesterol levels, raised triglyceride, and reduced HDL cholesterol levels). Clusters of three or more of these additional risk factors occur at four times the rate expected by chance. This clustering is attributed to an insulin resistance syndrome promoted by abdominal obesity. The amount of risk factor clustering accompanying elevated blood pressure was observed to increase with weight gain. Based on Framingham Study data the prevalence of insulin resistance syndrome in the general population could be as high as 22% in men and 27% in women. Risk of coronary disease, the most common and most lethal sequel to hypertension, increased stepwise with the extent of risk factor clustering. Among persons with hypertension, about 40% of coronary events in men and 68% in women are attributable to the presence of two or more additional risk factors. Only 14% of coronary events in hypertensive men and 5% of those in hypertensive women occurred in the absence of additional risk factors. Other important features of risk stratification of hypertension are the presence of an elevated heart rate and left ventricular hypertrophy, and an elevated fibrinogen that often accompany hypertension. Recent population-based data reported suggest that elevated renin accompanying hypertension may independently enhance the risk of coronary events. Because clustering of other major risk factors with hypertension is the rule, the prudent physician should routinely screen for the presence of these other factors. Multivariate risk assessment profiles are now available for coronary disease, stroke, peripheral artery disease, and heart failure, to enable physicians to pool all the relevant risk factor information so as to arrive at a composite risk estimate. Hypertensive patients are more appropriately targeted for therapy by such risk stratification and the goal of the therapy should be to improve the multivariate risk profile. PMID- 10678283 TI - Trials of antihypertensive treatment--new agenda for the millennium. AB - In 1968 a group of investigators from the US Veterans Association study reported that, compared to placebo, antihypertensive medication dramatically reduces morbidity and mortality in patients whose diastolic blood pressure was 115 mm Hg or higher. Three years later a reduction of morbidity had also been demonstrated in subjects whose diastolic pressure ranged from 95 to 114 mm Hg. In the following decades these findings were confirmed in numerous investigations of various populations. We now know that antihypertensive treatment is beneficial in mild hypertension, in isolated systolic hypertension, and in hypertension of the elderly. When it comes to the treatment of mild hypertension we will have to practice evidenceless medicine and act on the internal logic of the issue. The logic suggests that many present-day antihypertensive agents are harmless, that over the long term they may prevent target organ damage, that early intervention may be more beneficial than late treatment, and, finally, that treating mild hypertension may have a major positive impact on public health. PMID- 10678284 TI - Newly emerging pharmacologic differences in angiotensin II receptor blockers. AB - Several angiotensin II receptor blockers (ARB) are currently available for the treatment of hypertension. These drugs share a common mechanism of action antagonism of angiotensin II AT1 receptors; however, their receptor binding kinetics differ. Candesartan has a higher affinity for the AT1 receptor than all the other ARB. In addition, candesartan and irbesartan block the AT1 receptor with insurmountable antagonism, whereas losartan, valsartan, and eprosartan are competitive antagonists. The pharmacokinetics of these ARB also differ in terms of oral bioavailability, rate of absorption, metabolism, and route and rate of elimination. Both losartan potassium and candesartan cilexetil are prodrugs; however, losartan is partially converted into EXP3174 in the liver, whereas candesartan cilexetil is converted completely into candesartan during gastrointestinal absorption. On the basis of elimination half-lives, losartan, valsartan, and eprosartan may be classified as shorter acting and candesartan cilexetil and irbesartan as longer acting. Each drug effectively lowers blood pressure during once daily administration to patients with mild to moderate hypertension, with candesartan cilexetil requiring the lowest dosage and providing dose-dependent efficacy. Initial comparative clinical trials suggest that both candesartan cilexetil and irbesartan in the doses used are significantly more effective than losartan in lowering trough sitting diastolic blood pressure. It remains to be determined, however, whether the observed pharmacologic and pharmacokinetic differences among the members of the ARB class will have a clinically significant impact on long-term cardiovascular outcomes and reductions of cardiovascular mortality. PMID- 10678285 TI - Update on the clinical pharmacology of candesartan cilexetil. AB - The renin-angiotensin system plays a central role in the regulation of blood pressure through its primary effector hormone angiotensin II. Studies conducted nearly 30 years ago with peptidic angiotensin II receptor blockers (ARB) suggested that disruption of the renin-angiotensin system offered considerable promise for the treatment of hypertension as well as heart failure. This promise was initially realized with the advent of angiotensin converting enzyme inhibitors, and more recently with nonpeptidic ARB that selectively antagonize the AT1-angiotensin receptor subtype. The potent and long-acting agent candesartan cilexetil illustrates how these new ARB fulfill the promises suggested by the early studies. Candesartan cilexetil provides a clinically relevant, dose-dependent reduction in diastolic and systolic blood pressure at doses of 4 to 16 mg once daily in patients with mild to moderate hypertension. Recent studies suggest that further blood pressure lowering is obtained with a 32 mg once daily dose. In comparative clinical trials, 8 mg of candesartan cilexetil and 10 to 20 mg of enalapril provided comparable antihypertensive effects. The safety and tolerability profile of candesartan cilexetil is comparable to placebo. Notably, this agent does not produce the dry, nonproductive cough that often limits use of angiotensin converting enzyme inhibitors, nor does it cause side effects that limit other antihypertensive drug classes. On the basis of the results of initial clinical studies, ARB also possess cardioprotective and renoprotective properties that promise to expand the role that these new agents will play in treating cardiovascular disorders. PMID- 10678286 TI - Localization and function of angiotensin AT1 receptors. AB - The distributions of angiotensin AT1 and AT2 receptors have been mapped by in vitro autoradiography throughout most tissues of many mammals, including humans. In addition to confirming that AT1 receptors occur in sites known to be targets for the physiologic actions of angiotensin, such as the adrenal cortex and medulla, renal glomeruli and proximal tubules, vascular and cardiac muscle and brain circumventricular organs, many new sites of action have been demonstrated. In the kidney, AT1 receptors occur in high density in renal medullary interstitial cells. The function of these cells, which span the interstitial space between the tubules and the vasa rectae, remains to be determined. Renal medullary interstitial cells possess receptors for a number of vasoactive hormones in addition to AT1 receptors and this, in concert with their anatomic location, suggests they may be important for the regulation of fluid reabsorption or renal medullary blood flow. In the heart, the highest densities of AT1 receptors occur in association with the conduction system and vagal ganglia. In the central nervous system, high AT1 receptor densities occur in many regions behind the blood-brain barrier, supporting a role for neurally derived angiotensin as a neuromodulator. The physiologic role of angiotensin in many of these brain sites remains to be determined. The AT2 receptor also has a characteristic distribution in several tissues including the adrenal gland, heart, and brain. The role of this receptor in physiology is being elucidated, but it appears to inhibit proliferation and to participate in development. Thus, receptor-binding studies, localizing the distribution of AT1 and AT2 receptors, provide many insights into novel physiologic roles of angiotensin. PMID- 10678287 TI - AT1 receptors: coronary flow and flow reserve. AB - Angiotensin II, through its effects at the angiotensin-type 1 receptor, elevates arterial pressure and exacerbates hypertensive heart disease. Alterations in coronary hemodynamics, including reductions in coronary blood flow and flow reserve promotes coronary insufficiency and contributes to the increased cardiovascular risk associated with these conditions. In spontaneously hypertensive rats, coronary flow reserve, the difference between basal coronary blood flow and the flow achieved during maximal coronary vasodilation achieved by dipyridimole, was increased to a greater extent after treatment for 3 months with an angiotensin II receptor blocker as compared with an angiotensin converting enzyme inhibitor. The combination of the two agents, in equidepressor doses, almost restored coronary flow reserve to levels seen in normotensive Wistar Kyoto rats. This finding suggests a possible advantage of combination angiotensin converting enzyme inhibitors and angiotensin II receptor blocker therapy in patients with hypertension and hypertensive heart disease. PMID- 10678289 TI - The role of pharmacists in the detection, management, and control of hypertension: a national call to action. PMID- 10678288 TI - Renal responses to AT1 receptor blockade. AB - Because of the importance of the renin-angiotensin system in the pathophysiology of hypertension and in mediating associated alterations in renal function, angiotensin II (Ang II) AT1 receptor blockers provide a direct means of protecting against influences of excessive Ang II levels. The kidney is an important site of action of Ang II AT1 receptor blockers because intrarenal Ang II not only vasoconstricts the renal vasculature but also reduces sodium excretion and suppresses the pressure natriuresis relationship. Even in normal conditions, intrarenal Ang II content is greater than can be explained on the basis of circulating Ang II and is compartmentalized with proximal tubule concentrations of Ang I and Ang II being several times higher than plasma concentrations. The localization of angiotensinogen in proximal tubule cells further supports the concept that the proximal tubule secretes Ang II or precursors of Ang II into the tubular fluid to activate luminal Ang II receptors. Recent immunohistochemical studies have demonstrated an abundance of AT1 receptors on the luminal surface of proximal and distal tubule cells as well as on vascular smooth muscle cells of afferent and efferent arterioles and on glomerular mesangial cells. Activation of luminal AT1 receptors stimulates the sodium hydrogen exchanger and increases reabsorption rate. The prominence of AT1 receptors in vascular and epithelial tissues in the kidney provides the basis for the powerful effects of AT1 receptor blockers on renal function especially in hypertensive conditions. In the two-kidney, one-clip (2K1C) Goldblatt hypertensive rat model, the nonclipped kidney is renin depleted but the intrarenal Ang II levels are not suppressed and Ang II concentrations in proximal tubular fluid remain high (10(-8) mol/L). AT1 receptor blockers such as candesartan have been shown to cause significant increases in glomerular filtration rate, renal blood flow and proportionately much greater increases in sodium excretion and fractional sodium excretion. Ang II blockade also markedly increases the slope of the pressure natriuresis relationship. The collective actions of Ang II blockers on tubular transport and renal hemodynamics provide long-term effects to regulate sodium balance, which contributes to the long-term control of hypertension. PMID- 10678290 TI - Levalbuterol nebulizer solution: is it worth five times the cost of albuterol? AB - Albuterol is a 50:50 mixture of R-albuterol, the active enantiomer, and S albuterol, which appears to be inactive in humans. The Food and Drug Administration recently approved levalbuterol, the pure R-isomer, as a preservative-free nebulizer solution. Published studies indicate that it is neither safer nor more effective than an equimolar dose of racemic albuterol (levalbuterol 1.25 mg = albuterol 2.5 mg). However, these studies were conducted in patients with stable asthma (at the top of the dose-response curve), whereas a nebulized bronchodilator most likely would be used by patients with an acute exacerbation. Because such patients, in the hospital setting, often require higher doses of albuterol, the manufacturer's recommended dose of levalbuterol is likely to be too low for rescue therapy. Levalbuterol may cost as much as 5 times more than racemic albuterol, depending on purchase method. We conclude that levalbuterol offers no advantage over albuterol but is likely to be more costly. PMID- 10678291 TI - Pharmacologic, pharmacokinetic, and therapeutic differences among angiotensin II receptor antagonists. AB - Over the past 4 years, six angiotensin II receptor antagonists (ARBs) were approved for treating essential hypertension. They differ with respect to dosing, metabolism, elimination, clinical efficacy, and investigational applications. Candesartan cilexetil is the only prodrug among the agents. Losartan is distinguished from other ARBs by cytochrome P450 (CYP) 3A4- and CYP2C9-mediated biotransformation to its active metabolite EXP-3174. No ARB requires dosage adjustment for renal impairment, but the initial dose of losartan should be reduced 50% in hepatically impaired patients. None of the drugs is significantly cleared by hemodialysis. Completion of continuing trials will elucidate the drugs' role in treating heart failure, cerebral stroke, and myocardial infarction. PMID- 10678292 TI - Treatment of pediatric hypertension. AB - We conducted a MEDLINE search from January 1966-March 1999 to obtain information on clinical trials of treatment of pediatric hypertension. An article was selected for review if it described a randomized or nonrandomized study; randomized studies were given priority. Case reports were considered when studies were unavailable. Review articles were useful in identifying references. According to data we collected, hypertension is present in 1-3% of the pediatric population. Nonpharmacologic treatment may be effective initially in those with mild to moderate disease or as an adjunct to drug therapy. Drugs for treatment of chronic hypertension include calcium channel blockers, angiotensin-converting enzyme inhibitors, diuretics, and beta-blockers. Patient and drug characteristics determine therapy. Intravenous labetalol, nicardipine, and nitroprusside are effective for treating hypertensive emergencies. PMID- 10678293 TI - Human immunodeficiency virus drug resistance testing: state of the art in genotypic and phenotypic testing of antiretrovirals. AB - Antiretroviral drugs have significantly reduced death rates from the acquired immunodeficiency syndrome in the United States. They are highly effective in reducing viral replication, but their utility is threatened by rapid development of drug resistance. Although antiretroviral drug resistance testing is available by either genotyping or phenotyping, no consensus guidelines have been published regarding the appropriate use or interpretation of these new tests. Even though their role in clinical practice is not defined, it is important for clinicians to become familiar with relative advantages and disadvantages of genotypic and phenotypic testing and various mechanisms of antiretroviral resistance. PMID- 10678294 TI - A review of pathophysiology and therapy of patients with vasovagal syncope. AB - Vasovagal syncope is a common disorder that can compromise quality of life and lead to significant morbidity. It is characterized by an initial exaggerated sympathetic output followed by parasympathetic activation and sympathetic withdrawal, as shown by diagnostic head-up tilt (HUT) table testing. Numerous drugs have been evaluated for treating this disorder. beta-Blockers are well studied and commonly administered but are specifically more efficacious in patients with isoproterenol HUT than in those with regular HUT. The role of the serotonergic system has captured new interest. Selective serotonin reuptake inhibitors show promising results in preventing vasovagal syncope in treatment refractory patients. Also, new investigations suggest that serotonin receptor antagonism may be beneficial. Despite these findings, definitive treatment does not exist. PMID- 10678295 TI - Aerosolized antimicrobial therapy in acutely ill patients. AB - Recent data are sparking renewed interest in therapy with aerosolized antimicrobials in critically ill patients as well as other populations such as those with neutropenia, human immunodeficiency virus infection, and cystic fibrosis. Pneumonia is a common complication in these patients and is associated with substantial morbidity and increased mortality. Clinical trials evaluated aerosolized antimicrobials for the prevention and treatment of pneumonia in hospitalized patients. In addition, factors that affect the pulmonary deposition of aerosolized drugs in mechanically ventilated patients were identified. PMID- 10678296 TI - Evaluation of the influence of diabetes mellitus on antipyrine metabolism and CYP1A2 and CYP2D6 activity. AB - STUDY OBJECTIVE: To evaluate the metabolism of antipyrine, a general metabolic probe, caffeine, a probe for cytochrome P450 (CYP) 1A2 and N-acetyltransferase activity, and dextromethorphan, a specific probe for CYP2D6 activity in patients with type 1 or 2 diabetes mellitus. DESIGN: Prospective, controlled study. SETTING: Research facility. Patients. Fifteen patients with type 1 and 16 with type 2 diabetes, and 16 healthy controls. INTERVENTION: Each subject simultaneously received antipyrine 10 mg/kg, caffeine 100 mg, and dextromethorphan 30 mg. MEASUREMENTS AND MAIN RESULTS: The pharmacokinetics of antipyrine and its primary metabolites were determined from saliva and urine samples. Type 1 diabetes had marked effects on antipyrine metabolism whereas type 2 disease did not alter the metabolism of any of the probe drugs. The apparent oral clearance of antipyrine was increased 72% in patients with type 1 disease compared with controls (p=0.0001). In addition, formation clearances of 4 hydroxyantipyrine and 3-hydroxymethylantipyrine were increased by 74% and 137% in those patients relative to controls. The caffeine metabolic index (paraxanthine/caffeine) was increased 34% (p=0.11), and N-acetylation and CYP2D6 phenotype were not altered. CONCLUSION: The metabolism of antipyrine is increased in patients with type 1 diabetes. Based on in vitro reports of antipyrine metabolism and current caffeine metabolic index data, the predominant effect of type 1 diabetes appears to be an increase in CYP1A2 activity. Assessment of the effect of the disease on other specific CYP metabolic pathways is warranted. PMID- 10678297 TI - Quantification of human H1 histamine receptor mRNA from peripheral blood. AB - STUDY OBJECTIVE: To develop a reverse transcription (RT)-polymerase chain reaction (PCR) technique to detect and quantify human histamine1 (H1) receptor mRNA in peripheral blood. METHODS: Primer pairs were based on the human H1 receptor nucleotide sequence. A competitive reference standard (CRS) was developed that used the same primers as wild-type mRNA but contained a 92-bp deletion. RT-PCR was performed with 5 microg of total RNA obtained from venous blood of six subjects that was added to known concentrations of CRS RNA. Linear regression comparing wild-type with CRS product was used to quantify wild-type mRNA. MEASUREMENTS AND MAIN RESULTS: Three subjects had detectable H1 mRNA, with a range of 31-435 pg. In three subjects PCR product was not detected, although the presence of RNA was confirmed. Redesigned primer pairs showed mRNA to H1 receptor in two of the remaining subjects, but it was undetectable in the third. CONCLUSION: RT-PCR can be used to detect and quantify human H1 receptor mRNA from peripheral blood. PMID- 10678298 TI - Increased therapeutic failure for cephalexin versus comparator antibiotics in the treatment of uncomplicated outpatient cellulitis. AB - We reviewed records of outpatients to determine the therapeutic failure rate of cephalexin in treating uncomplicated cellulitis. Therapeutic failure was defined as an increase in antibiotic dosage, prescription renewal, or addition or substitution of another antibiotic. Demographics, physical characteristics, risk factors, intervention, and outcome data were collected. Twenty-seven percent of patients failed therapy with an oral antibiotic. The failure rate for cephalexin was 40% versus 20% for comparator antibiotics (p=0.02, odds ratio [OR] 2.62, 95% confidence interval [CI] 1.18-5.75). We identified no statistically significant variables related to cephalexin failure. Concomitant acid suppressive therapy was administered with cephalexin in 42% of failures and 20% of nonfailures (p=0.11, OR 2.78, 95% CI 0.77-9.87). These data suggest that cephalexin's efficacy was less than that of other antimicrobials in treating cellulitis, possibly related to concurrent acid suppression. PMID- 10678299 TI - A multidisciplinary renal clinic for corticosteroid-induced bone disease. AB - A multidisciplinary clinic to manage complicated bone disease was established due to the high prevalence of osteoporosis in corticosteroid-treated patients with a history of organ transplantation or chronic glomerulonephritis. Assessments were performed by a renal clinical pharmacist, nephrology nurse, and rheumatologist. Of 70 patients (27 men, 43 women) evaluated from December 1997-June 1999, 37% had osteoporosis (30% spine, 23% hip, 16% both sites) and 34% had a history of fracture. Analysis revealed low 1,25-hydroxyvitamin D3 levels (15 patients), hormone deficiency (16), elevated parathyroid hormone (27), and history of taking at least one other risk drug in addition to corticosteroids (58). Thirty-nine percent of patients had a documented height loss (mean 1.0 in.). Other risk factors included 32 episodes of graft rejection requiring additional corticosteroids, history of smoking (24 patients), poor physical activity (40), and low dietary calcium intake (47). Drug interventions included calcium and/or vitamin D (44 patients), calcitonin (7), alendronate (20), and hormone replacement therapy (11). Preliminary results showed an increase in bone mineral density (a surrogate marker for fracture risk) of 3-5%. An organized clinic to assess osteoporosis risks can unmask a large population of patients with documented bone loss. Appropriate interventions such as drug therapy and lifestyle changes may increase bone mineral density. A long-term benefit of therapy, although not measured in this study, may be a decreased predisposition to fractures and their sequelae. PMID- 10678300 TI - Impact of an indigent care program on use of resources: experiences at one hospital. AB - Thirty-six indigent patients hospitalized within 6 months of study initiation and who met criteria were enrolled in a 6-month assistance program to determine if provision of both medical care and prescription drugs at no cost would be associated with a change in overall health care charges secondary to a change in therapy adherence. A historical control group was identified. Charges for drugs and medical care, and the number of hospitalizations and emergency room visits were evaluated for comparison with the pretreatment period. Adherence to drug regimens was measured using the Med-Out indicator. Inpatient admissions decreased by 39.5% (from 43 to 26) and outpatient visits decreased by 64.4% (from 194 to 69). This amounted to a cost avoidance to the hospital of $378,183. The cost of drugs during the study was $27,588. Patients who adhered to therapeutic regimens provided an even greater cost avoidance. PMID- 10678301 TI - A database analysis of potentially inappropriate drug use in an elderly medicaid population. AB - We conducted a cross-sectional retrospective review of 1996 Kentucky Medicaid Pharmacy claims data to examine the prevalence of potentially inappropriate drug use in 64,832 Medicaid recipients aged 65 years and older who received a prescription. Twenty-seven percent of patients received at least one potentially inappropriate agent. Prevalence was higher for nursing home residents (33%) than for community dwellers (24%). Amitriptyline (7.6%), propoxyphene (6.5%), doxepin (4.0%), and indomethacin (2.3%) were the most prescribed potentially inappropriate agents. Education programs and interventions aimed at optimizing the prescribing and dispensing of the most appropriate drugs are needed. PMID- 10678302 TI - Phenytoin as a possible cause of acetaminophen hepatotoxicity: case report and review of the literature. AB - A 55-year-old woman was hospitalized for treatment of community-acquired pneumonia. Unexplained, moderate elevations in hepatic transaminase and enzyme levels prompted review of her drug regimen. She had taken acetaminophen 1,300 6,200 mg/day during the hospitalization. She also received phenytoin for posttraumatic seizures. Acetaminophen was discontinued, and the patient's liver chemistries returned to normal within 2 weeks of discharge. Acetaminophen is metabolized in part by cytochrome P450 (CYP) 2E1, and inducers of CYP2E1 are known to predispose patients to acetaminophen-related hepatotoxicity. Phenytoin induces CYP2C and CYP3A4 isoforms, but not CYP2E1. The literature suggests, however, that CYP3A4 may participate in acetaminophen metabolism to a greater extent than previously realized, and induction of this isoform may predispose patients to acetaminophen-induced hepatotoxicity. PMID- 10678303 TI - Potentiation of oral anticoagulation and hemarthrosis associated with nabumetone. AB - Concomitant therapy with warfarin and nonsteroidal antiinflammatory drugs (NSAIDs) is of concern due to the potential for increased bleeding. Nonsteroidal antiinflammatory drugs may alter patient response to warfarin by pharmacodynamic or pharmacokinetic interactions. A man receiving long-term, stable warfarin therapy experienced a significant increase in international normalized ratio 1 week after nabumetone was added to his regimen. Despite prompt reduction of the warfarin dosage, he experienced hemarthrosis of his right knee. Previous reports suggested lack of interaction between nabumetone and warfarin. Caution and close monitoring are advisable when the two agents are administered concomitantly. PMID- 10678304 TI - Epistaxis associated with elevation of INR in a patient switched to generic warfarin. AB - A 61-year-old man with atrial fibrillation receiving Coumadin brand warfarin was switched to Barr brand warfarin without his knowledge as a result of a retail pharmacy dispensing error. The patient took the same dosage for 6-7 days and experienced severe epistaxis that required two visits to the emergency room. Previously, his coagulation values were within therapeutic range, but when tested at the emergency room the international normalized ratio was elevated. The patient denied changes in therapy compliance, diet, alcohol ingestion, or use of other drugs. His only other drug, taken periodically, was sildenafil for erectile dysfunction. Clinicians should be aware of differences between branded and generic compounds. PMID- 10678305 TI - Psychiatric diagnosis in sociocultural context. PMID- 10678306 TI - International systems of diagnosis in psychiatry. AB - International standardization of diagnosis is the culmination of developments set in motion in Europe during the first half of the 19th century. Its ultimate rationale has been the science of descriptive psychopathology. The enterprise implies that a common way of defining, describing, identifying, naming, and classifying such disorders is possible and that a common system of psychiatric diagnosis constitutes a first step toward dealing with them. Its natural science approach implies that social and cultural factors are extraneous. Yet, as discussed, the enterprise is based upon unique historical and cultural responses to human behavioral individuality. Cultural aspects of psychiatric phenomena create tensions in the application of the internationalist enterprise: Although in theory applicable to all people regardless of populational/genetic, national, or cultural background, it is used by clinicians of highly specific cultural origin and in settings characterized by distinctive cultural traditions about sickness, healing, nonsickness or health, and social behavior. Tensions created by the international enterprise are discussed and illustrated by drawing attention to how cultural factors impact on its basic assumptions and by a selective review of literature. PMID- 10678307 TI - Painful coitus: a review of female dyspareunia. AB - The literature on female dyspareunia is reviewed with an emphasis on description and classification, incidence and prevalence, etiological factors, and treatment approaches. The research is found to be plagued with methodological problems and characterized by a persistent dichotomization of issues into physiological and psychological categories. A systematized and integrated approach to the study of coital pain is proposed. PMID- 10678308 TI - Visual hallucinations in patients from an ophthalmology clinic and medical clinic population. AB - Although visual hallucinations have been associated with patients with visual disorders, no study has specifically examined whether visual hallucinations are indeed more prevalent than in a general medical population. In this study, 127 consecutive visual disorder patients and 100 consecutive general medical patients were screened for complex visual hallucinations. A total of 6.3% of visual disorder patients and 2% of general medical patients had visual hallucinations. Interestingly, the two medical patients with visual hallucinations also had visual disorders. Factors significantly associated with visual hallucinations were female sex (p = .029) and lower cognitive score (p = .001). Data from a previous study of patients with the visual disorder age-related macular degeneration were combined with this study to increase the sample size of visual hallucinators. Factors significantly associated with visual hallucinations in the combined sample were female sex (p = .015), living alone (p = .019), having hearing problems (p = .047), older age (p = .013), and lower cognitive score (p < .001). Implications and future research are discussed. PMID- 10678309 TI - Longitudinal, expert, all data procedure for psychiatric diagnosis in patients with psychoactive substance use disorders. AB - The Longitudinal, Expert, All Data (LEAD) procedure has been proposed as a criterion for the assessment of the procedural validity of diagnostic instruments. The authors evaluated the procedure's test-retest reliability and whether it enhanced diagnosis based on a single interview. Data were collected using interviews and questionnaires to assess current and lifetime substance use disorders and common comorbid disorders in 100 patients recruited from a substance abuse treatment program. An initial diagnostic interview was conducted by a primary expert who, at the end of treatment, formulated LEAD diagnoses for each patient, based on the results of the research assessment and all available clinical records. Secondary experts used a similar procedure in a subsample of 40 patients to provide a measure of test-retest reliability. Overall reliability of LEAD diagnoses was good, though it varied from excellent for substance use disorders to poor for some comorbid psychiatric disorders. As a consequence of the LEAD procedure, the total number of substance use diagnoses increased significantly, with no effect on diagnostic reliability. Based on these findings, we conclude that, while the overall reliability of the LEAD procedure is comparable to other diagnostic methods, there was considerable variability among groups of diagnoses. The additional cost of the procedure would appear to be justified only when diagnostic sensitivity is at a premium and principally for the diagnosis of psychoactive substance use disorders. Before it is widely used as a diagnostic criterion measure, the utility of the LEAD procedure should be evaluated in a variety of patient samples and under varying circumstances. PMID- 10678310 TI - A sibling study of sensation seeking and opiate addiction. AB - Substance abuse and its correlative personality traits may have familial associations. We assessed the relationship between sensation seeking and drug use in 201 opiate addicts and 133 of their siblings in a cross-sectional family study. Probands and their drug-abusing siblings showed greater sensation seeking than their non-drug-abusing siblings and this diagnosis accounted for the most variance in regression models. Degree of sensation seeking correlated among drug abusing siblings and with age of first drug use. The results are discussed in terms of substance abuse typologies and for using sensation-seeking assessments for prevention and treatment of substance abuse. PMID- 10678311 TI - Remission and relapse in subjects with panic disorder and panic with agoraphobia: a prospective short-interval naturalistic follow-up. AB - This article reports on the course of uncomplicated panic disorder and panic with agoraphobia on 309 patients participating in the Harvard/Brown Anxiety Research Project, a prospective longitudinal study of patients with DSM-III-R-defined anxiety disorders. At 1 year, there was a .39 probability of full remission for uncomplicated panic disorder and a .17 probability of full remission for panic disorder with agoraphobia Similar differences in time to remission for these syndromes were still found when criteria for remission were made less stringent. However, even requiring less improvement for remission left a large percentage of subjects in an episode, and for those that remitted, relapse occurred quickly, indicating a chronic and recurrent course of illness. This is the first longitudinal, prospective, naturalistic study on a large cohort of subjects with anxiety disorders to have regular, structured, short-interval follow-up. Our results are consistent with the view that panic disorder has a chronic course with high rates of relapse after remission and longer episodes when agoraphobia is a part of the constellation of symptoms. PMID- 10678312 TI - A prospective, follow-along study of the course of social phobia: II. Testing for basic predictors of course. AB - This study examined the 65-week outcome of a group of subjects with social phobia to determine predictors of course. Social phobic patients in the Harvard/Brown Anxiety Disorders Research Project study were followed for 65 weeks using the Longitudinal Interval Follow-Up Evaluation-UpJohn scale. The following variables did not predict outcome over the course of the study: sex, age of onset, duration of illness, lifetime history of various anxiety disorders, current comorbidity of anxiety or depressive disorders, Global Assessment Scale score, or measures of role functioning. We find that in a social phobic population with a mean duration of illness of 18 years, none of the tested variables examined predicted 65-week outcome. PMID- 10678313 TI - Hyperreligiosity in psychotic disorders. PMID- 10678314 TI - The heterogeneity of stealing behaviors in Chinese patients with anorexia nervosa in Hong Kong. PMID- 10678315 TI - Domains of psychopathology: an approach to the reduction of heterogeneity in schizophrenia. AB - The manifest clinical heterogeneity of schizophrenia, combined with the failure, to date, to demonstrate the existence of a unitary disease process, has led to the conceptualization of schizophrenia as a pathophysiologically heterogeneous disorder. Various approaches have been developed to define homogeneous subgroups of schizophrenic patients. An alternative approach to the use of multiple criteria for defining putative disease entities is the use of specific sign and symptom complexes, or domains of psychopathology, for reducing heterogeneity. There is now considerable evidence supporting the separation of schizophrenic symptoms into three domains: hallucinations and delusions, thought disorder, and deficit symptoms. The conceptual evolution and validating evidence for this approach are reviewed, and an illustration of how the domains of psychopathology are applied in schizophrenia research is presented. PMID- 10678316 TI - Application of artifical intelligence principles to the analysis of "crazy" speech. AB - Artificial intelligence computer simulation methods can be used to investigate psychotic or "crazy" speech. Here, symbolic reasoning algorithms establish semantic networks that schematize speech. These semantic networks consist of two main structures: case frames and object taxonomies. Node-based reasoning rules apply to object taxonomies and pathway-based reasoning rules apply to case frames. Normal listeners may recognize speech as "crazy talk" based on violations of node- and pathway-based reasoning rules. In this article, three separate segments of schizophrenic speech illustrate violations of these rules. This artificial intelligence approach is compared and contrasted with other neurolinguistic approaches and is discussed as a conceptual link between neurobiological and psychodynamic understandings of psychopathology. PMID- 10678317 TI - Dissecting psychotic speech. PMID- 10678318 TI - Metaphors and models in the neuropsychiatry of language. PMID- 10678319 TI - Artificial intelligence and schizophrenic speech analysis. PMID- 10678320 TI - The problem of missing clinical data for research in psychopathology: some solution guidelines. AB - There are no guidelines to help psychiatric researchers statistically adjust for missing data. We discuss the problems resulting from missing values, and illustrate some of them with examples from our work. Using structured instruments, we obtained clinical information from 241 patients. Some instrument items were not rated, and these did not occur randomly: hallucinations and delusions were most frequently unrated, especially in chronic schizophrenics, and patients with high scores for other psychopathology. Systematically assigning an intermediate value between present and absent to nonrated items was a satisfactory solution, unaffected by nonrandom missing values. This simple solution was equivalent to a complicated one (vectoring) in discriminating patients. When relationships between variables are linear, we recommend the intermediate value method as a practical solution to missing values. We stress that missing values do not mean missing information, and the most common response to missing values (dropping subjects) is least informative. PMID- 10678321 TI - Are there two depressive syndromes after stroke? AB - The objective of this study was to describe the clinical characteristics of minor depression after stroke and to compare this disorder with poststroke major depression and the nondepressed state. Ninety-four stroke inpatients were examined 8 weeks after stroke and reexamined 15 months later. Twenty-one (22%) of the 94 patients suffered from minor depression, 14 (15%) suffered from major depression, and 59 (63%) were not depressed. Minor depressed patients were twice as symptomatic as nondepressed patients but were only half as symptomatic as major depressed patients. Minor depressed patients were more likely than nondepressed patients to have a previous history of stroke and were more physically disabled. They were less likely than major depressed patients to have a family history of affective disorder. Depression symptom severity was associated with greater physical disability among minor but not major depressed patients. Fewer minor than major depressed patients were depressed at 15 months. These findings suggest that poststroke major and minor depression may be different depressive syndromes. Some cases of minor depression may be construed as an adjustment reaction to stroke disability. PMID- 10678322 TI - Group differences in the relationship between apathy and depression. AB - The purpose of this study was to evaluate the discriminability of apathy and depression by determining whether the relationship of these two dimensions of behavior varies in different diagnostic groups. Using the authors' Apathy Evaluation Scale and the Hamilton Rating Scale for Depression, we rated 123 subjects, mean age 72 years, who met research criteria for healthy elderly controls, left or right hemisphere stroke, probable Alzheimer's disease, and major depression. Elevated apathy scores unassociated with elevated depression were most frequent in Alzheimer's disease and right hemisphere stroke, and also occurred in a small number of left hemisphere stroke and normal subjects. In major depression, apathy was associated with high depression scores, although a substantial number of major depressives showed elevated depression without elevated apathy. In left hemisphere stroke, probable Alzheimer's disease, and major depression, there were significant positive correlations between apathy and depression. The slope of the regression of apathy on depression was greatest in probable Alzheimer's disease and major depression. These results indicate that the relationship between apathy and depression differs across diagnostic groups and, thus, support the discriminability of apathy and depression. PMID- 10678323 TI - Impulsivity scores in patients with obsessive-compulsive disorder. PMID- 10678324 TI - Propranolol: a treatment for pseudoakathisia. PMID- 10678325 TI - Monitoring chronic fatigue syndrome. PMID- 10678326 TI - A case of posttraumatic stress disorder masked by pseudoseizures in a Jewish Iranian immigrant in Israel. PMID- 10678327 TI - Posttraumatic stress disorder and sociopolitical context: a comment. PMID- 10678328 TI - Comparing buprenorphine and methadone maintenance. PMID- 10678329 TI - Aspirin for atrial fibrillation. PMID- 10678330 TI - Preventing stroke in patients with atrial fibrillation. PMID- 10678331 TI - Bed rest ineffective as therapy. PMID- 10678332 TI - Helicobacter pylori eradication for nonulcer dyspepsia is of limited value. PMID- 10678334 TI - Interspeciality differences in medical resource utilization. PMID- 10678335 TI - Physician satisfaction with Medicaid managed care: the Missouri experience. AB - BACKGROUND: Medicaid managed care is important to health reform at the state level. However, little is known about physician satisfaction with these programs. We sought to measure this satisfaction in Missouri and determine its predictors. METHODS: We surveyed a random sample of primary care physicians participating in Medicaid managed care (n = 670) or traditional Medicaid (n = 670). Primary outcomes measured were physicians' satisfaction Medicaid managed care, traditional Medicaid and commercial managed care. Satisfaction was measured on a 5-point Likert-type scale. RESULTS: The response rate was 52%. Physicians participating in Medicaid managed care were less likely to be satisfied or very satisfied with Medicaid managed care (28.6%) than with commercial managed care (40%) or their previous experience with traditional Medicaid (39.7%). Among physicians participating in traditional Medicaid, 29.8% were satisfied or very satisfied with traditional Medicaid. Physicians participating in Medicaid managed care were less satisfied with clinical autonomy under that system in comparison with their previous experience with traditional Medicaid (relative difference = 10.8%, P =.001). In multiple linear regression analyses, clinical autonomy (R2 = 0.40) was a strong predictor of overall satisfaction with Medicaid managed care. CONCLUSIONS: Enhancing physicians' clinical autonomy may result in improved satisfaction with Medicaid managed care. State Medicaid agencies should include physician satisfaction as a measure of Medicaid managed care plans' quality. PMID- 10678336 TI - A circle, broken. PMID- 10678337 TI - Alcohol-related discussions in primary care: a report from ASPN. Ambulatory Sentinel Practice Network. AB - BACKGROUND: Problem drinking is common, and a 15-minute intervention can help some patients reduce drinking to safe levels. Little is known, however, about the frequency and duration of alcohol-related discussions in primary care. METHODS: Nineteen clinicians in the Ambulatory Sentinel Practice Network (ASPN) collected data about alcohol-related discussions for 1 week following their usual office routine (Phase 1) and for 1 week with the addition of routine screening for problem drinking (Phase 2). Of those, 15 clinicians collected data for a third week after receiving training in brief interventions with problem drinkers (Phase 3). Clinicians collected data on standard ASPN reporting cards. RESULTS: In Phase 1 the clinicians discussed alcohol during 9.6% of all visits. Seventy-three percent of those discussions were shorter than 2 minutes long, and only 10% lasted longer than 4 minutes. When routine screening was added (Phase 2), clinicians were more likely to discuss alcohol at acute-illness visits, but the frequency, duration, and intensity of such discussions did not change. Only 32% of Phase 2 discussions prompted by a positive screening result lasted longer than 2 minutes. After training, the duration increased (P <.004). In Phase 3, 58% of discussions prompted by a positive screening result lasted longer than 2 minutes, but only 26% lasted longer than 4 minutes. CONCLUSION: Routine screening changed the kinds of visits during which clinicians discussed alcohol use. Training in brief-intervention techniques significantly increased the duration of alcohol related discussions, but most discussions prompted by a positive screening result were still shorter than effective interventions reported in the literature. PMID- 10678338 TI - Use of a high-sensitivity rapid strep test without culture confirmation of negative results: 2 years' experience. AB - BACKGROUND: Optimal diagnostic management of patients with pharyngitis is controversial. In our study, we compared streptococcal complication rates at a large suburban medical center during 2 time periods: when pharyngitis patients were managed almost exclusively with throat culture and when they were managed primarily with a high-sensitivity antigen test without culture confirmation of negative results. METHODS: Using a combination of Current Procedural Terminology and International Classification of Diseases, Ninth Revision codes, we studied all patients seen for either pharyngitis or known streptococcal complications during a 4-year period. We then reviewed all available charts of patients with known streptococcal complications for coding accuracy. We compared streptococcal complication rates during each -time period. RESULTS: A total of 30,036 patients were seen for pharyngitis during the 4 years. A streptococcal diagnostic test was used in 66% of patient encounters. During the first 2 years (period 1), 99.9% of the tests ordered were blood agar plate throat cultures. During the second 2 years (period 2), 76.6% of tests ordered were high-sensitivity antigen tests without culture confirmation of negative results. Suppurative complications occurred in 37 patients in period 1 and 36 patients in period 2. There were no cases of acute rheumatic fever in either period. There was one case of poststreptococcal glomerulonephritis in period 2. CONCLUSIONS: Use of a high sensitivity antigen test without culture confirmation of all negative results has not been associated with an increase in suppurative and nonsuppurative complications of group A beta-hemolytic streptococci. PMID- 10678339 TI - The impact of regular vaginal pH screening on the diagnosis of bacterial vaginosis in pregnancy. AB - BACKGROUND: Bacterial vaginosis has recently been associated with preterm labor and delivery. The purpose of our study was to determine whether regular prenatal vaginal pH testing resulted in more frequent diagnoses of bacterial vaginosis and other vaginal infections, more frequent treatment with antibiotics, and fewer preterm deliveries. We also sought to determine the sensitivity and specificity of pH testing and vaginal symptom reporting in identifying vaginal infections. METHODS: Our study was a prospective clinical trial involving 121 pregnant women randomized to receive either standard prenatal care, including routine inquiry about vaginal symptoms, or standard care supplemented by vaginal pH testing. Women with symptoms or a vaginal pH level >4.5 received a wet mount examination. Confirmed infections were treated according to study protocols. RESULTS: Women who received regular pH testing showed significantly higher detection rates for bacterial vaginosis than controls (48.4% vs 27.1%, P =.015) and more frequent detection of Trichomonas vaginalis (7.8% vs 1.7%, P = .116). A higher percentage of women in the experimental group were treated for bacterial vaginosis and trichomoniasis (46.9% vs 27.1%, P =.024), and the preterm birth rate was one half that of the control group (4.7% vs 10.2%, P = .243). The presence of vaginal symptoms or a vaginal pH level >4.5 identified bacterial vaginosis or trichomoniasis with 84.4% sensitivity. CONCLUSIONS: In our study, frequent vaginal pH testing during pregnancy resulted in more frequent diagnosis and treatment of bacterial vaginosis. Since vaginal symptoms and elevated pH levels appear to be useful in screening for bacterial vaginosis and trichomoniasis, frequent pH testing should be evaluated in larger studies. PMID- 10678340 TI - The evidence regarding the drugs used for ventricular rate control. AB - OBJECTIVE: Our goal was to determine what drugs are most efficacious for controlling the ventricular rate in patients with atrial fibrillation. SEARCH STRATEGY: We conducted a systematic review of the literature published before May 1998, beginning with searches of The Cochrane Collaboration's CENTRAL database and MEDLINE. SELECTION CRITERIA: We included English-language articles describing randomized controlled trials of drugs used for heart rate control in adults with atrial fibrillation. DATA COLLECTION/ANALYSIS: Abstracts of trials were reviewed independently by 2 members of the study team. We reviewed English-language abstracts of non-English-language publications to assess qualitative consistency with our results. MAIN RESULTS: Forty-five articles evaluating 17 drugs met our criteria for review. In the 5 trials of verapamil and 5 of diltiazem, heart rate was reduced significantly (P <.05), both at rest and with exercise, compared with placebo, with equivalent or improved exercise tolerance in 6 of 7 comparisons. In 7 of 12 comparisons of a beta-blocker with placebo, the beta-blocker was efficacious for control of resting heart rate, with evidence that the effect is drug specific, as nadolol and atenolol proved to be most efficacious. All 9 comparisons demonstrated good heart rate control with beta-blockers during exercise, although exercise tolerance was compromised in 3 of 9 comparisons. In 7 of 8 trials, digoxin administered alone slowed the resting heart rate more than placebo, but it did not significantly slow the rate during exercise in 4 studies. The trials evaluating other drugs yielded insufficient evidence to support their use, but those drugs may yet be promising. CONCLUSIONS: The calcium-channel blockers verapamil or diltiazem, or select beta-blockers are efficacious for heart rate control at rest and during exercise for patients with atrial fibrillation without a clinically important decrease in exercise tolerance. Digoxin is useful when rate control during exercise is less a concern. PMID- 10678341 TI - Becoming an information master: using POEMs to change practice with confidence. Patient-Oriented Evidence that Matters. AB - Today's physicians are faced with identifying, evaluating, and applying a huge quantity of medical information. In addition, many stakeholders in health care, from patients to payers, are taking an active role in the previously inviolable process of physician decision making. This is the third paper in a series discussing the concept of information mastery. In the first paper we discussed using the criteria for Patient-Oriented Evidence that Matters (POEMs) to distill clinically relevant information. The second paper in the series focused on techniques for efficiently obtaining this information from the myriad resources available. In this paper we discuss the final step in the process, changing practice habits after finding new information that necessitates it. We discuss managing change, consider barriers, and present ideas to help with the process. PMID- 10678342 TI - Depression diagnoses and antidepressant use in primary care practices: a study from the Practice Partner Research Network (PPRNet). AB - BACKGROUND: We examined the pharmacologic management and follow-up of adults with newly diagnosed depression, and the use of antidepressants among patients not diagnosed with depression in primary care practice. A total of 389 physicians in 39 practices in the Practice Partner Research Network (PPRNet), a national network of primary care physicians provided data for the study. METHODS: We performed a retrospective cohort study for the year 1996 using demographic, contact, diagnosis, and prescription data available in the December 1997 PPRNet database. We identified patients with new diagnoses of depression from the problem lists in the electronic medical record. Psychopharmacologic agents prescribed within 5 days of the diagnosis, follow-up contacts within 6 months of the diagnosis, and diagnoses of patients prescribed antidepressants without a new diagnosis of depression were also identified. We performed descriptive analyses for all practices and for individual practices. RESULTS: During 1996, there were 149,327 active adult patients in the 39 participating practices. Of the 131,141 patients without a history of depression or antidepressant prescription, 2103 (1.6%) had a new diagnosis of depression in 1996. Incidence among the 39 practices ranged from 0.4% to 4.0%. Forty-nine percent of the newly diagnosed patients received an antidepressant prescription within 5 days of diagnosis; 81% of the prescriptions were for selective serotonin reuptake inhibitors. Ninety percent of the patients prescribed antidepressants had at least one contact in the 6 months after diagnosis (mean = 5.3 contacts). One third of the patients who had not begun antidepressants within 5 days of their diagnoses started taking one by the end of 1996. Among the 149,327 active patients, 6.3% received a prescription for an antidepressant in 1996. More than 40% of these patients had never been diagnosed with depression. CONCLUSIONS: Our study highlights the high prevalence and wide interpractice variations of diagnosing depression and prescribing antidepressants in primary care. Follow-up of patients newly diagnosed with depression was common and consistent with published guidelines. Opportunities for increased detection and treatment of depression exist in approximately half of the study practices. PMID- 10678343 TI - High prevalence of overweight children in Michigan primary care practices. An UPRNet study. Upper Peninsula Research Network. AB - BACKGROUND: Our goal was to determine whether the prevalence of obesity in children who receive care in Michigan primary care practices is greater than national and state prevalences. METHODS: We compared prevalences of overweight children and adolescents in primary care practices with the results of the National Health Examination Survey (NHES), the National Health and Nutrition Examination surveys, and a contemporary survey of Michigan schoolchildren. We collected data from 19 rural family practice offices and 2 urban clinics. We measured the heights and weights of 993 consecutive patients aged 4 to 17 years who visited one of the participating practices during the spring of 1996. RESULTS: Obesity prevalences were the main outcome measure. Of the boys, 38% were above the 85th percentile of the NHES, and 16% were above the 95th percentile. Of the girls, 33% were above the 85th percentile, and 13% were above the 95th percentile. Prevalences of obesity were much higher among the primary care patients than in the results of the national surveys and the contemporary Michigan schoolchildren survey. CONCLUSIONS: The prevalences of obesity for children and adolescents presenting for care in Michigan primary care practices are higher than the prevalences documented in state and national surveys. A larger systematic study is needed to confirm or refute these findings. If this prevalence of obesity in primary care patients is confirmed, explanations for the differences should be explored. PMID- 10678344 TI - Helicobacter pylori eradication does not improve symptoms of nonulcer dyspepsia. PMID- 10678345 TI - Neuroleptics for behavioral symptoms of dementia. PMID- 10678346 TI - Treating carpal tunnel syndrome. AB - BACKGROUND CTS is a common problem caused by compression of the median nerve at the wrist resulting in hand numbness, loss of dexterity, muscle wasting, and decreased functional ability at work. This study investigated the efficacy of a corticosteroid injection just proximal (not into) the carpal tunnel for CTS. POPULATION STUDIED: Study participants included 60 patients referred to a neurology clinic in Amsterdam, Netherlands, with CTS symptoms for longer than 3 months' duration and confirmed with electrophysiological tests. Patients in the intervention and control groups had symptoms for an average of 32 months and 25 months, respectively. In patients with bilateral symptoms, the arm with the most severe symptoms was chosen for randomization. Patients aged younger than 18 years and those who had previous treatment for CTS were excluded. STUDY DESIGN AND VALIDITY: Patients were randomized to receive an injection of either lignocaine (Lidocaine 10 mg) and methylprednisolone 40 mg or a lignocaine 10-mg injection only. The site of injection was proximal to the carpal tunnel on the volar side of the forearm 4 cm proximal to the wrist crease, between the tendon of the radial flexor muscle and the long palmar muscle. Injections were given at a 10 degrees to 20 degrees angle with a 3-cm needle. At baseline, there were no significant differences between the control group and the intervention group. The study was performed at one clinic where one neurologist performed all injections. Thus, we do not know if the results of this technique can be consistently reproduced. No patients were reported lost to follow-up at 1 year. To ensure blinding of the treatment assignment, a pharmacist wrapped the syringes in paper and a second neurologist performed outcomes assessment interviews. One month after the initial injection, patients were asked whether they had no symptoms or only minor symptoms that they considered so much improved that they felt no further treatment was necessary. Investigators broke the trial code at follow-up assessment visits to offer nonresponders an injection with methylprednisolone or surgery. OUTCOMES MEASURED: Patients were considered improved if they self reported no symptoms or only minor symptoms needing no additional treatment. Other symptoms (weakness, nighttime pain) or impact on lifestyle and occupation were not reported. RESULTS: At 1 month, 20% of the patients in the control group had improved compared with 77% of patients in the intervention group (P <.001; number needed to treat = 1.8). After 1 year, 8 of the 23 patients (35%) who initially responded to methylprednisolone required a second injection. A total of 86% of nonresponders in the control group improved after receiving a methylprednisolone injection, but 50% of these patients went on to need surgical treatment within 1 year. The investigators reported no side effects to the injection. PMID- 10678347 TI - Treatment modalities for primary basal cell carcinomas. PMID- 10678348 TI - The hidden value of primary care. PMID- 10678349 TI - Treatment of influenza. PMID- 10678350 TI - Targeting gene expression to tumor cells with loss of wild-type p53 function. AB - The tumor suppressor protein p53 is a transcription factor that can positively regulate the expression of critical target genes involved in negative control of cell growth or induction of apoptosis; p53 is also able to suppress the transcription of other genes by virtue of its ability to bind components of the basal transcription machinery. Over 50% of human tumors are characterized by p53 mutations that result in a loss of wild-type p53 (wtp53) function in the transcriptional control of these target genes. We have exploited this loss of p53 function in the regulation of gene transcription to develop a novel gene therapy strategy that maximizes expression of the potential therapeutic gene in tumors while simultaneously down-regulating the same gene in normal cells. In one construct (unit I), the potential therapeutic gene (in this case represented by a luciferase reporter) is placed under the control of a promoter such as the heat shock protein 70 gene promoter, which is repressed by wtp53 but overexpressed in many tumor cells with defective p53 function. Residual expression of the reporter in normal cells is repressed by cotransfection of another construct (unit II) consisting of a repressor of unit I under the control of a promoter that is activated by wtp53 expression. Unit II contains a promoter with a consensus wtp53 binding site driving a transcriptional repressor or an antisense construct for the gene in unit I. Our results suggest that this dual control approach may represent a strategy with wide applications in the field of cancer gene therapy. PMID- 10678351 TI - Modifications of the fiber in adenovirus vectors increase tropism for malignant glioma models. AB - Recombinant adenovirus (Ad) vectors provide a means of local, therapeutic gene delivery to a wide range of neoplasms. Ad-mediated gene therapy trials in malignant glioma models have been limited by the need for high viral titers and multiple dosages. In an attempt to improve Ad vector gene transfer, we studied human (U87, D54) and rodent (GL261, C6) malignant glioma cell lines transfected with various doses of unmodified Ad vectors (AdZ), Ad vectors that contain an alteration of the fiber-coat protein and that direct virus binding to heparan sulfate receptors (AdZ.F(pK7)), and Ad vectors with modifications of the fiber coat protein that direct virus binding to alpha1, integrin cellular receptors (AdZ.F(RGD)). AdZ.F(pK7) increased the frequency of cells expressing the reporter gene, beta-galactosidase, and improved transduction by 2- to 20-fold compared with AdZ in U87, D54, and GL261 cells. In U87, D54, GL261, and C6 tumors, AdZ.F(pK7) increased gene transfer by 10- to 100-fold compared with AdZ. AdZ.F(RGD) increased gene expression in C6 xenografts compared with AdZ, but had reduced transduction compared with the C6 xenografts of AdZ in all other glioma tumors. These findings suggest that the increased tropisms resulting from alterations of the Ad vector fiber-coat protein as in AdZ.F(pK7) and AdZ.F(RGD) offer a feasible approach to improving in vitro and in vivo transduction efficiencies in certain malignant glioma cell lines. PMID- 10678352 TI - Cytosine deaminase suicide gene therapy for peritoneal carcinomatosis. AB - Gene therapy is a novel therapeutic approach that might soon improve the prognosis of some cancers. We investigated the feasibility of cytosine deaminase (CD) suicide gene therapy in a model of peritoneal carcinomatosis. DHD/K12 colorectal adenocarcinoma cells transfected in vitro with the CD gene were highly sensitive to 5-fluorocytosine (5-FC), and a bystander effect could also be observed. Treating CD+ cells with 5-FC resulted in apoptosis as detected by terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick-end labeling. In vitro, several human cell lines derived from ovarian or colorectal carcinomas, as well as the rat glioblastoma 9 L cell line, responded to CD/5-FC and showed a very strong bystander effect. 5-FC treatment of peritoneal carcinomatosis generated in syngeneic BDIX rats by CD-expressing DHD/K12 cells led to a complete and prolonged response and to prolonged survival. Our study thus demonstrated the efficacy of CD suicide gene therapy for the treatment of peritoneal carcinomatosis. PMID- 10678353 TI - Highly efficient suicide gene expression in hepatocellular carcinoma cells by epstein-barr virus-based plasmid vectors combined with polyamidoamine dendrimer. AB - The present study was aimed at devising an efficient nonviral strategy for suicide gene therapy of hepatocellular carcinoma (HCC). To improve the efficiency of DNA delivery and expression, we applied Epstein-Barr virus (EBV)-based plasmid vectors instead of conventional plasmid vectors and combined them with cationic liposome (EBV/lipoplex) or polyamidoamine dendrimer (PAAD) (EBV/polyplex). When the beta-galactosidase gene was transferred to HuH7, PLC/PRF/5, or HLE cells, < or =50-fold higher beta-galactosidase activities were demonstrated in the cells transfected with EBV vector compared with those transfected with conventional plasmid vectors. PAAD-mediated transfection of HCC with pSES.Tk (an EBV-based vector carrying the herpes simplex virus-1 thymidine kinase gene) resulted in a marked reduction in viable cell number by the addition of ganciclovir (GCV). The HCC cells transfected with pSES.Tk/PAAD showed 100- to 1000-fold higher susceptibilities to GCV than those transfected with pS.Tk (a conventional plasmid vector carrying herpes simplex virus-1 thymidine kinase gene)/PAAD. The pSES.Tk transfected HCC cells were effectively killed by day 9 in culture with a clinically feasible concentration of GCV (25 microM), whereas the pS.Tk transfected cells survived the culture. These results demonstrate highly efficient suicide gene transfer into various HCC cells by EBV-based plasmid vectors in vitro, suggesting the possible application of this nonviral vector system to gene therapy of HCC. PMID- 10678354 TI - Adenoviral transfer of xenogeneic MHC class I gene results in loss of tumorigenicity and inhibition of tumor growth. AB - The immune system confers protection against a variety of pathogens and contributes to the destruction of neoplastic cells. Foreign major histocompatibility complex (MHC) protein serves as a potent stimulus to the immune system. In this report, a mouse H-2Kb gene was introduced into two poorly immunogenic tumor cell lines, a mouse colonic carcinoma cell line, MCA-26 (H 2Kd), and a rat mammalian carcinoma cell line, LN-4, in an effort to stimulate tumor rejection. Our results showed that the expression of xenogeneic MHC class I antigen completely abolished the LN-4 tumorigenicity in rats, whereas the expression of allogeneic MHC class I antigen only partially reduced the MCA-26 tumorigenicity in mice. Rats with tumor regression of LN-4/H-2Kb developed a T helper type 1-dominant response, whereas rats with LN-4 tumor growth developed a T helper type 2-dominant response. The immunized rats that experienced LN-4/H-2Kb tumor regression further developed protective immunity against a subsequent challenge of LN-4 cells. This protective immunity was mediated by the LN-4 tumor specific cellular immune response against both the transduced and the parental LN 4 cells. Recombinant adenoviral vectors are highly efficient at in vitro and in vivo gene delivery. The LN4 cells transfected with the recombinant adenovirus AdV H-2Kb in vitro expressed the cell surface H-2Kb molecule by fluorescence activated cell sorter analysis. Adenovirus-mediated H-2Kb gene transfer in vivo can further significantly inhibit pre-established LN-4 tumors. Those rats with complete tumor regression further developed protective immunity against the subsequent challenge of a parental LN-4 tumor. Therefore, our study indicates that the adenovirus-mediated transfer of xenogeneic MHC class I gene may be an effective alternative to the current protocol of cancer gene therapy in which the allogeneic MHC class I gene is used. PMID- 10678355 TI - Potentiation of ganciclovir toxicity in the herpes simplex virus thymidine kinase/ganciclovir administration system by ponicidin. AB - The herpes simplex virus thymidine kinase (HSV-TK)/ganciclovir (GCV) administration system is commonly used in gene therapy trials. We have evaluated the effect of ponicidin, a diterpenoid isolated from a plant, Rabdosia ternifolia, on the cell-killing activity of the anti-herpes drugs acyclovir (ACV) and GCV. Ponicidin preferentially activated HSV-1-specific TK but not cellular kinases. In HSV-infected cells, ponicidin significantly accumulated the phosphorylated metabolites of GCV and suppressed the extracellular release of GCV. These data suggested that the cytotoxicities of ACV and GCV in HSV-TK expressing cells might be potentiated by ponicidin. After transfected with the HSV-1 TK gene, COS-1 and several human cancer cells became highly sensitive to the cytotoxic properties of the nucleoside analogs. When ponicidin at the concentration without antiviral activities (0.2 microg/mL) was combined with ACV or GCV, the cytotoxic levels in HSV-TK-expressing cells were enhanced by 3- to 87 fold and 5- to 52-fold, respectively, compared with the nucleoside alone. When the stability of the bioactivity of ponicidin in the blood of mice was evaluated, the substance showed relatively long-lasting effects on the potentiation of the anti-herpetic and cytotoxic activities of GCV after intravenous administration. These data suggest that the combined use of ponicidin with GCV will be effective for cancer gene therapy, because high cytotoxicity in viral TK-expressing cells should yield more rapid and enhanced tumor elimination. PMID- 10678356 TI - Fusogenic effects of murine retroviruses and cationic enhancers of transduction. AB - The maturation of retrovirus particles involves proteolytic cleavage of the envelope glycoprotein transmembrane component, resulting in conversion of the virus particle to a fusogenic or infectious state. Susceptible murine cells exposed to virus-containing supernatants from ecotropic retroviral helper cells occasionally fused to neighboring cells, resulting in syncytia (giant cells with multiple nuclei). Polycationic molecules dramatically enhanced the effect, leading to widespread cell death. The degree of cell fusion was dependent upon the retroviral envelope subtype (ecotropic-->amphotropic, gibbon ape leukemia virus was negative) as well as on the polycationic reagent used (G9 dendrimer- >Lipofectamine-->polybrene). Cell fusion effects were not mediated by the retroviral vector backbone, because virus-containing supernatants from helper cells (without vector) and vector producer cells had a similar effect. Human target cells were not fused by any type of murine retrovirus; in addition, amphotropic virus from human helper cells was not fusogenic toward murine cells. Thus, fusogenic effects were important during the propagation of vectors using murine helper cells but were not a significant factor during the transduction of human cells. PMID- 10678357 TI - Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells. AB - Poor transfection efficiency is the major drawback of lipofection. We showed previously that addition of transferrin (TF) to Lipofectin enhanced the expression of a reporter gene in HeLa cells by 120-fold and achieved close to 100% transfection efficiency. The purpose of this study was to determine whether TF and other ligands could improve the efficiency of lipofection in lung carcinoma cells. Confluent A549, Calu3, and H292 cells were transfected for 18 hours with a plasmid DNA (pCMVlacZ) using Lipofectin plus TF, insulin, or epidermal growth factor as the vector. The transfected cells were assessed for transfection efficiency by beta-galactosidase activity (light units/microg protein) and the percentage of blue cells following 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside staining. Lipofectin supplemented with epidermal growth factor yielded the largest enhancement of lipofection efficiency (< or =23-fold over that by Lipofectin alone) in all three cell lines. Insulin significantly enhanced the lipofection efficiency in A549 and Calu3 cells but not in H292 cells, whereas TF showed significant lipofection efficiency-enhancing effect in Calu3 and H292 cells but not in A549 cells. The transfection efficiency correlated well with the amounts of DNA delivered to the nucleus as well as the amounts of the receptor. These results indicate that the gene delivery strategy employing ligand-facilitated lipofection can achieve high transfection efficiency in human lung carcinoma cells. In addition, enhancement of the expression of the receptor may be a possible strategy for increasing the efficiency of gene targeting. PMID- 10678358 TI - Vaccinia as a vector for tumor-directed gene therapy: biodistribution of a thymidine kinase-deleted mutant. AB - Tumor-directed gene therapy, such as "suicide gene" therapy, requires high levels of gene expression in a high percentage of tumor cells in vivo to be effective. Current vector strategies have been ineffective in achieving these goals. This report introduces the attenuated (thymidine kinase (TK)-negative) replication competent vaccinia virus (VV) as a potential vector for tumor-directed gene therapy by studying the biodistribution of VV in animal tumor models. A TK deleted recombinant VV (Western Reserve strain) expressing luciferase on a synthetic promoter was constructed. Luciferase activity was measured in vitro after transduction of a variety of human and murine tumor cell lines and in vivo after intraperitoneal (i.p.) delivery in C57BL/6 mice with 7-day i.p. tumors (10(6) MC-38 cells). Three other in vivo tumor models were examined for tumor specific gene expression after intravenous delivery of VV (human melanoma in nude mice, adenocarcinoma liver metastasis in immunocompetent mice, and subcutaneous sarcoma in the rat). In addition, a replication-incompetent vaccinia (1 microg of psoralen and ultraviolet light, 365 nm, 4 minutes) was tested in vitro and in vivo and compared with active virus. Luciferase activity in i.p. tumors at 4 days after i.p. injection of VV was >7000-fold higher than lung, >3000-fold higher than liver, and >250-fold higher than ovary. In addition, intravenous injection of VV resulted in markedly higher tumor luciferase activity compared with any other organ in every model tested (up to 188,000-fold higher than liver and 77,000-fold higher than lung). Inactivation of the virus resulted in negligible gene expression in vivo. In summary, VV has a high transduction efficiency in tumor cells with high levels of gene expression. The results suggest a selective in vivo replication of TK-deleted VV in tumor cells. Replication competent, TK deleted VV appears to be an ideal vector for testing the in vivo delivery of toxic genes to tumor cells. PMID- 10678359 TI - In vivo efficacy and toxicity of 5-fluorocytosine/cytosine deaminase gene therapy for malignant gliomas mediated by adenovirus. AB - We evaluated the therapeutic efficacy and neurotoxicity of adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for experimental malignant brain tumors. The 5-FC sensitivity in 9 L cells infected by an adenovirus vector expressing CD (AdexCACD) was increased 1700-fold compared with control cells. Rats bearing 9 L brain tumors were treated with an intratumoral injection of AdexCACD followed by intraperitoneal administration of 5-FC. The rats demonstrated remarkable inhibition of tumor growth by magnetic resonance imaging, and 7 of 10 rats survived for >90 days. To evaluate the potential side-effects of the 5-FC/CD gene therapy, rats were treated with an intracerebral injection of AdexCACD into the right basal ganglia and with 5-FC. The magnetic resonance imaging showed a highly enhanced area on the gadollinium enhanced T1-weighted image at 18 days postinjection. Pathologically, this corresponded to an area of necrosis with surrounding apoptotic cells. In addition, there was demyelination and gliosis with enlargement of the lateral ventricles. These results suggest that the 5-FC/CD gene therapy may provide an anticancer effect for malignant brain tumors in humans, but also show that there are neurotoxic effects on normal brain tissue. PMID- 10678360 TI - Effectiveness of cancer vaccine therapy using cells transduced with the interleukin-12 gene combined with systemic interleukin-18 administration. AB - We introduced the interleukin-12 (IL-12) gene into mouse renal cell carcinoma (RenCa) cells to develop a tumor vaccine and to examine mechanisms of tumor rejection. IL-12-secreting RenCa (RenCa/IL-12) cells were completely rejected when implanted into syngeneic BALB/c but not athymic nude mice, suggesting that T cells were involved in this antitumor effect. Depletion of natural killer (NK) cells in nude mice did not affect the tumor growth of RenCa/IL-12. The simultaneous injection of mitomycin C-treated RenCa/IL-12 inhibited the tumor growth of parental RenCa injected at a distant site, whereas injection of mitomycin C-treated parental RenCa did not. The antitumor effect of RenCa/IL-12 as a cancer vaccine was induced by CD8+ T cells and NK cells and was inhibited by CD4+ T cells. Although the systemic administration of recombinant IL-18 (rIL-18) alone did not inhibit the tumor growth, it did enhance the cancer vaccine effect of RenCa/IL-12. The combination therapy of RenCa/IL-12 and the systemic administration of rIL-18 retarded even the growth of established tumors. The effector cells of this combination therapy consist not only of CD8+ T cells and NK cells but also of CD4+ T cells. This synergistic cancer vaccine effect of in situ secretion of IL-12 and the systemic administration of rIL-18 may be attributed to a functional change of CD4+ T cells. PMID- 10678361 TI - Attenuating the growth of tumors by intratumoral administration of DNA encoding Pseudomonas exotoxin via cationic liposomes. AB - A gene therapy approach was taken to inhibit tumor growth by transfecting tumor cells with a plasmid encoding a truncated but active form of Pseudomonas exotoxin A (PE), using cationic lipids as the transfection reagent. Cells transfected with this plasmid express PE intracellularly and undergo apoptosis. Transfection was optimized in vitro using two cationic lipids, DOGS and DOSPER. A ratio of between 1:4 and 1:10 (wt/wt) was found to be optimal for DOSPER, and the ratio 1:4 was used for the in vivo study when a smaller injection volume was desired. Estimating the activity of the PE-encoding plasmid was done both directly, by counting cells in vitro after transfection, and by using a cytotoxicity assay, and indirectly, by cotransfecting the plasmid with a plasmid carrying a reporter beta-galactosidase gene and observing a reduction in beta-galactosidase activity with increasing amounts of the PE-encoding plasmid. The cotransfection method was found to be very sensitive, and showed transfection of cells even with 1-2 ng of the PE-encoding plasmid per 10(5) cells. Complexes of the PE-encoding plasmid together with cationic lipid were injected into tumor xenografts in athymic nude mice. The tumor growth of transfected tumors was attenuated compared with control untreated tumors or tumors transfected with a nontoxin-expressing vector. These results indicate the potential of such a treatment for attenuating solid tumor growth in vivo. PMID- 10678362 TI - The nitroreductase/CB1954 combination in Epstein-Barr virus-positive B-cell lines: induction of bystander killing in vitro and in vivo. AB - Epstein-Barr virus (EBV)-based gene delivery vectors that preferentially express toxic genes in EBV-infected cells could be used to target EBV-positive tumors for destruction. We have shown previously that the cytosine deaminase (CD) enzyme, which converts the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5 fluorouracil efficiently kills EBV-positive cells in the presence of 5-FC, with a substantial bystander killing effect in vitro and in vivo. To identify the optimal enzyme/prodrug combination for treating EBV-positive lymphomas, we have compared the effectiveness of the CD/5-FC combination with the nitroreductase (NTR)/CB1954 combination for killing EBV-positive B-cell lines. NTR metabolizes CB1954 into an alkylating agent that cross-links DNA. When the CD gene or the NTR gene were transfected into two different EBV-positive B-cell lines in vitro, approximately 90% of cells were killed in a prodrug-dependent manner, although the transfection efficiency was <5%. However, severe combined immunodeficient mouse tumors containing either 30% or 100% of NTR-expressing Burkitt lymphoma (Jijoye) cells were growth inhibited, but not cured, by treatment with intraperitoneal CB1954 (20 mg/kg/day) for 10 days. These results suggest that the NTR/CB1954 combination induces efficient bystander killing of EBV-positive B-cell lines in vitro but may not be as effective as the CD/5-FC combination for treating B-cell lymphomas in vivo. PMID- 10678363 TI - Comparison of the genotoxic and apoptosis-inducing properties of ganciclovir and penciclovir in Chinese hamster ovary cells transfected with the thymidine kinase gene of herpes simplex virus-1: implications for gene therapeutic approaches. AB - We studied the genotoxic and apoptosis-inducing properties of ganciclovir (GCV) and penciclovir (PCV) using Chinese hamster ovary cells stably transfected with the thymidine kinase (tk) gene of herpes simplex virus-1 (HSV-1). Cells expressing HSVtk were 300 and 100 times more sensitive than their isogenic HSVtk- counterparts to the cytotoxic effects of GCV and PCV, respectively. Using radiolabeled drugs, GCV was found to be incorporated into the genomic DNA much more effectively than PCV. GCV was highly potent in inducing chromosomal aberrations compared with PCV, which provoked less sister chromatid exchanges and chromosomal changes using equimolar or equitoxic doses. For both agents, apoptosis was shown to be the major route of cell killing. Time course experiments revealed that neither genotoxicity nor apoptosis were induced within the cell cycle exposed to the drug; they are late events provoked in the following cell cycle(s). This indicates that the incorporation/exposure step of GCV or PCV into DNA is not decisive for triggering genotoxicity and apoptosis, but that events occurring subsequently, presumably during replication of a DNA containing the nucleotide analogs, are of major importance. Because PCV, unlike GCV, induced highly effectively apoptosis without exerting much genotoxicity, the use of PCV as a relatively safe alternative drug for suicide gene therapy of malignant diseases is recommended. PMID- 10678364 TI - Purified herpes simplex thymidine kinase retroviral particles. II. Influence of clinical parameters and bystander killing mechanisms. AB - High-titer, purified herpes simplex virus thymidine kinase (HSV-TK) retroviral particles, followed with intraperitoneal ganciclovir (GCV) were tested in Fischer rats bearing 1-week established 9L gliosarcomas. 9L cells were infused intracranially through a cannula on day 0, given intracranial infusions of HSV-TK retroviral particles on days 7-12, and given 5 or 10 daily doses of intraperitoneal GCV starting at day 14. Tumor volumetric studies performed on rat brains at day 26 after tumor infusion revealed significant differences in experimental groups receiving HSV-TK retroviral particles plus 10-day GCV or the 100% transduced 9L-TK group receiving GCV versus control groups treated with either buffer, HSV-TK vector, or either 5- or 10-day regimens of GCV alone. The duration of GCV administration and the volume of tumor burden influenced efficacy. Adjuvant dexamethasone did not significantly affect efficacy. Experiments in which 9L rechallenge of animals treated with 9L-TK/GCV or 9L tumors treated with HSV-TK vector/GCV indicated that a host immune response was involved in rejecting the rechallenge tumor. Outcome was dependent upon the site and size of the rechallenge inoculum. In vitro, bystander effects were significant but were especially marked when cell-to-cell contact was maintained. Cumulatively, the data indicate that both the bystander effect and the host immune response are responsible for the efficacy observed in the suicide gene therapy paradigm using purified HSV-TK retroviral particles and GCV to treat smaller tumor burden in rats with 9L gliosarcoma. PMID- 10678365 TI - Ligand-mediated cytolysis of tumor cells: use of heregulin-zeta chimeras to redirect cytotoxic T lymphocytes. AB - New specificities may be engrafted onto lymphocytes by the transfer of genes for chimeric receptors that combine antigen recognition and signal-transducing elements. We have engineered and evaluated a new class of chimeric receptors that use the natural ligands of receptors found to be frequently overexpressed by cancer cells. The heregulin molecule, a ligand for Her3 and Her4 receptors when fused with the CD3 zeta-chain, was capable of redirecting T lymphocytes to recognize and respond to cancer cell lines that overexpress these receptors. Thus, CD8+ T lymphocytes were isolated from a healthy individual and transduced to express the chimeric heregulin-zeta receptor. These modified effector cells acquired the ability to specifically lyse a breast cancer cell line that overexpresses Her3 and Her4. A new class of chimeric receptors, such as heregulin zeta, endowing anti-cancer effector cells with the potential to recognize and eliminate tumor targets, are likely to increase the effectiveness of adoptive immunotherapy for the treatment of cancer. PMID- 10678366 TI - Development of a murine orthotopic model of leukemia: evaluation of TP53 gene therapy efficacy. AB - The onco-suppressor gene TP53 has potential use in the gene therapy of many human cancers including leukemias. The latter indication derived from numerous experimental reports of p53-mediated suppressing effects on human and murine leukemia cells in vitro. However, few in vivo experiments have been performed, and those that have used a subcutaneous injection of p53-transduced leukemia cells. Thus, we developed an orthotopic leukemia model in adult, syngenic mice to evaluate the feasibility of TP53-mediated therapeutic approaches. We found that among other cells, v-src-transformed 32D myeloid progenitors induce leukemia when injected intravenously in syngenic mice. The resulting malignancy resembles the clinical manifestations of human acute myeloid leukemia because it is characterized by a massive invasion of bone marrow compartments, splenomegaly, generalized lymphadenopathy, and a macroscopic or microscopic infiltration of the kidneys, liver, and lungs. When these 32Dv-src cells were infected with a TP53 recombinant retrovirus before intravenous injection, we found a decreased mortality and, in those animals that develop leukemia, a drastic reduction of the generalized organ infiltration, suggesting that exogenous TP53 expression might be used for ex vivo bone marrow purging from leukemia cells. PMID- 10678367 TI - Immunogene therapy against mouse leukemia using B7 molecules. AB - B7 costimulatory molecules play an important role in T-cell activation. It is well known that tumor cells that express B7 molecules can elicit antitumor immunity, but little is known regarding which B7 molecule, B7-1 (CD80) or B7-2 (CD86), can do so more efficiently. To address this issue, we have introduced B7 1 or B7-2 into 8709 cells, a radiation-induced mouse myelocytic leukemic cell line, and have compared their potentials regarding the induction of antitumor immunity. Either B7-1- or B7-2-transduced monoclonal sublines, 8709/B7-1 or 8709/B7-2, respectively, diminished tumorigenicity in syngeneic C3H mice. Some reports have indicated that B7-1 is superior to B7-2 in the induction of antitumor immunity. Contrary to these results, the 8709/B7-2 lines are superior to the 8709/B7-1 lines in their capacity to induce antitumor immunity. In vivo depletion of lymphocyte subsets demonstrated that both CD4+ and CD8+ T cells were indispensable for B7-1- or B7-2-dependent antitumor immunity, whereas natural killer cells were not. These results suggest that in some circumstances, B7-2 molecule is more effective than B7-1 molecule in eliciting antitumor immunity. PMID- 10678368 TI - Tetracycline-induced expression of an anti-c-Myb single-chain antibody and its inhibitory effect on proliferation of the human leukemia cell line K562. AB - Ablation of c-Myb function might be an effective approach for the therapy of chronic myelogenous leukemia or other c-myb-dependent malignancies. To this end, we have previously used an intracellular anti-c-Myb single-chain antibody (sFv) to achieve the functional knockout of the c-Myb oncoprotein. In this study, we have employed a tetracycline-inducible system to control the expression of the sFv. A nuclear-localizing form of an anti-c-Myb sFv was cloned into a tet regulated plasmid vector. Using a transient expression system in COS-1 cells, we observed that doxycycline (Dox) induced expression of the sFv in a dose-dependent manner, and that the sFv was localized mainly in the nucleus. The Dox-induced anti-c-Myb sFv also inhibited the transactivating activity of c-Myb in a dose dependent manner. We subsequently confirmed the Dox-induced expression of the sFv in the leukemia cell line K562. Proliferation of the target leukemia cells was also inhibited. These results suggest that the anti-c-Myb sFv may represent a viable method for gene therapy of c-myb-dependent hematopoietic malignancies. PMID- 10678369 TI - The aortic valve blood supply. AB - BACKGROUND AND AIM OF THE STUDY: Normal valves are known to be metabolically active, yet the route of oxygen delivery is unclear. Although diffusion from the valve surface is the presumed source, the presence, distribution and importance of the aortic valve's vascular bed is unclear. METHODS: Seventeen porcine hearts (51 cusps) were obtained at slaughter. The coronary circulation was pressure rinsed with a heparin solution and filled with an Aquablack solution at physiological pressure. The cusps were subsequently dissected and fixed for viewing with an inverted microscope. The anterior leaflet of the mitral valve was evaluated as a control for the vessel filling protocol. Using Adobe Photoshop and captured images, whole cusps were reconstructed, a grid was overlaid and vascular distribution evaluated. RESULTS: Of the 15 porcine aortic valves that filled, 32/45 (71%) of the cusps contained vessels. Nine valves had vasculature in all three cusps and two valves were completely avascular. Vessels were found predominantly in the basal third of the cusps and extended in from the commissures almost to the level of the free edge. There was a statistically significant difference (p<0.001) between the appearance of vessels in the basal region and in the mid and free edge regions of the valve. No difference in vascularization pattern was noted between the left, right or non-coronary cusps. CONCLUSIONS: The presence of a vasculature suggests that the metabolic activity of the cusp is greater than can be supported by diffusion from the cusp surface alone. The absence of functioning vessels might explain the failure associated with cryopreserved implants and may play a role in the durability problems faced by bioprosthetic valves. It will also be important to consider the role of this intrinsic circulation with the advent of tissue-engineered valves. PMID- 10678370 TI - Significant increase of aortic root volume and commissural area occurs prior to aortic valve opening. AB - BACKGROUND AND AIM OF THE STUDY: The increased use of autologous, homologous or heterologous aortic root demands a detailed knowledge of its anatomy and function. The advent of 3-D digital sonomicrometry offered the opportunity to acquire precise information on the root and leaflet movements during the cardiac cycle. METHODS: Under cardiopulmonary bypass, sonomicrometry crystals were implanted in the aortic root and valve of eight sheep. Crystals were sutured at each commissure (n = 3), the top of the sinotubular junction (n = 3), lowest point of the annulus (n = 3), and leaflet tip (n = 3). 3-D coordinates of each crystal were recorded, together with left ventricular and aortic root pressures and ECG. When the animal had returned to a stable hemodynamic condition, the maximum and minimum distances between two crystals, and areas between three crystals, were calculated. Changes in root volume and leaflet position were time related to the pressure changes. RESULTS: The most significant change between maximum and minimum distance between crystals during the cardiac cycle occurred at the commissural level. Similarly, the triangle defined by the three commissural crystals showed the greatest change in area (47%). The root volume increased by an average of 22%; about 40% of this increase occurred during the isovolumic phase. The aortic leaflets began to open before ejection. CONCLUSION: We postulate that aortic valve opening is initiated by the outward pull of the commissures. These findings should impact on aortic root surgery. PMID- 10678371 TI - Aortic root dilation prior to valve opening explained by passive hemodynamics. AB - BACKGROUND AND AIM OF STUDY: During the June 1999 World Symposium on Heart Valve Disease, the mechanism by which the aortic valve opens was discussed. It was suggested, indirectly, that the recently discovered contractile elements within the aortic valve may be responsible. We propose that aortic root dilation does not require any active mechanism within the leaflets or aortic wall, and provide an explanation based entirely on the passive hemodynamics of the aortic valve and root. METHODS AND RESULTS: Previous studies using cine fluoroscopy and sonomicrometry have reported a 5-7% expansion of the aortic root during ventricular contraction, prior to aortic valve opening. Simplified force calculations indicate that the mechanical interactions between the aortic valve and root produce an inward pull on the commissures, constraining the aorta from fully dilating. During systole, as the pressure in the ventricle increases and the aortic valve becomes unloaded, the inward pull on the commissures is reduced and the aorta is able to dilate fully. Aortic dilation therefore occurs before the aortic valve opens. CONCLUSION: We conclude that this mechanism of aortic root dilation prior to valve opening is purely passive, and does not require any active process by the aortic valve or the aortic root. PMID- 10678372 TI - Decalcification of the aortic valve does not prevent early recalcification. AB - BACKGROUND AND AIM OF THE STUDY: The excellent results with atrioventricular valve reconstruction have stimulated surgeons to attempt reconstruction of calcified aortic valves using decalcifying techniques, but long-term results have been disappointing. The aim of this in vitro study was to evaluate the surface structure of decalcified aortic valve tissue and its potential for recalcification. METHODS: Aortic leaflets were removed from 26 patients with aortic stenosis during elective valve replacement and decalcified by meticulous dissection. Representative specimens were prepared for scanning electron microscopy (SEM) and calcium content in the heavily calcified part of the leaflet in both macroscopically non-calcified and decalcified tissue was determined by atomic absorption spectroscopy (AAS). Additional probes of 'non-calcified' and decalcified tissue were incubated for two and four weeks with medium containing a physiological concentration of calcium to determine their potential for recalcification. As a control, 13 specimens from non-calcified valves were incubated according to the same protocol. RESULTS: All calcified specimens contained high calcium levels (142.70+/-53.76 mg/g). Surgical dissection reduced tissue calcium content significantly (10.04+/-13.43 mg/g). Following two weeks' incubation with calcium, these specimens retained significantly higher levels of calcium (2.88+/-5.17 mg/g) than the 'non-calcified' specimens (19.17+/-7.61 versus 13.49+/-6.27 mg/g; p<0.05); after four weeks similar calcium levels were reached (32.00+/-10.27 versus 28.35+/-9.84 mg/g; p = NS). Non-calcified tissue showed the lowest calcium uptake (4.75+/-4.55 mg/g and 12.29+/-9.43 mg/g at two and four weeks; p<0.05). SEM revealed a loss of endothelial coverage in the calcified areas; decalcification led to an irregular fibrillar surface. Only parts of the macroscopically normal tissue contained endothelial cells, whereas the control tissue showed intact cellular coverage. CONCLUSION: Aortic valve decalcification can effectively remove calcifications, but leaves a fibrillar structure that tends rapidly to accumulate calcium. Even normal-appearing tissue from diseased valves has a higher potential for calcification than normal valvular tissue. These data support the observation of only limited clinical benefits being derived after aortic valve decalcification for aortic stenosis. PMID- 10678373 TI - Mechanics of cryopreserved aortic and pulmonary homografts. AB - BACKGROUND AND AIM OF THE STUDY: The surgical placement of pulmonary valve grafts into the aortic position (the Ross procedure) has been performed for three decades. Cryopreserved pulmonary valves have had mixed clinical results, however. The objectives of this study were to compare the mechanics of cryopreserved human aortic and pulmonary valve cusps and roots to determine if the pulmonary root can withstand the greater pressures of the aortic position. METHODS: Six aortic and six pulmonary valve roots were obtained from the Oxford Valve Bank. They were harvested during cardiac transplantation from hearts explanted for dilated cardiomyopathy (mean patient age 68 years). The whole roots were initially stored frozen at -186 degrees C, then shipped packed on dry ice. After complete thawing, the roots were pressurized whole; test strips were then cut from the valve cusps, roots and sinuses and tested for stress/strain, stress relaxation, and ultimate failure strength. RESULTS: The pulmonary roots were more distensible (30% versus 20% strain to lock-up) and less compliant when loaded to aortic pressures. The pulmonary valve cusp and root tissue also showed greater extensibility and greater stiffness (lower compliance) when subjected to the same loads. CONCLUSION: We conclude that mechanical differences between aortic and pulmonary valve tissues are minimal. The pulmonary root should withstand the forces imposed on it when placed in the aortic position. However, if implanted whole, the pulmonary root will distend about 30% more than the aortic root when subjected to aortic pressures. These geometric changes may affect valve function in the long term and should be appreciated when implanting a pulmonary valve graft. PMID- 10678374 TI - Results of the Ross operation in rheumatic versus non-rheumatic aortic valve disease. AB - BACKGROUND AND AIM OF THE STUDY: A total of 213 patients underwent the Ross operation at our institution between January 1990 and January 1999. Outcome was assessed in rheumatic (RH) patients and compared with that in patients with other etiology (non-RH). METHODS: After exclusion of 69 patients with a follow up of <18 months, the study group comprised 144 patients (119 RH, 25 non-RH). Patients were studied clinically and by echo-Doppler cardiography preoperatively, within 2 months and 6-8 months after surgery, and yearly afterwards. Preoperative assessment included age, gender, body surface area (BSA), type of aortic valve lesion and additional valve disease, left and right ventricular outflow tract (LVOT, RVOT) dimensions, and left ventricular (LV) size, function and mass. Postoperatively, patients were studied for presence and severity of autograft regurgitation, mitral regurgitation, LV size, function and mass, and incidence and timing of reoperation. RESULTS: On average, RH patients were older and had higher BSA, more aortic regurgitation than stenosis, more additional mitral valve disease (mostly regurgitation), larger LV size and poorer LV function. Mitral valve repair was performed in 24% of RH patients versus 0% of non-RH patients. Postoperatively, differences in LV size, function and mass remained present, but diminished during follow up. The autograft reoperation incidence was 22% (26/119) in RH patients versus 8% (2/25) in non-RH patients (p = NS). Preoperative predictors for reoperation in the RH group were severe concomitant mitral regurgitation (MR), followed by male gender and large indexed LVOT (all p<0.001 by discriminant analysis). CONCLUSION: Marked differences were present in patient characteristics between rheumatic and nonrheumatic patients who underwent the Ross operation. Rheumatic patients had a higher incidence of autograft reoperation. Severe concomitant MR was the most important predictor for reoperation in rheumatic patients. PMID- 10678375 TI - Comparative analysis of left ventricular hemodynamics and hypertrophy after aortic valve replacement with homografts or mechanical valves. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to examine comparatively the effects of prosthetic and homograft valves in the aortic position on ventricular hemodynamics and structure. METHODS: Hemodynamic evaluations were performed at rest and during exercise in 38 patients who had undergone aortic valve replacement (AVR) with either a homograft (n = 19) or prosthetic valve (19-23 mm; n = 19). Using echocardiographic, electrocardiographic and hematologic methods, the pressure gradient (PG); aortic valve area; diameters of left anterior wall, posterior wall (PW) and interventricular septum (IVS); ejection fraction (EF); left ventricular mass (LVM) and mass index (LVMI); electrocardiographic data of LV hypertrophy; hemoglobin; hematocrit and lactate dehydrogenase (LDH) levels were measured. RESULTS: LVM and LVMI decreased significantly after surgery in both groups (p<0.001), but the decrease was significantly greater in the homograft group (p<0.05). The IVS and PW diameters in the homograft group decreased significantly postoperatively (p<0.05); the inter-group difference was also significant (p<0.01). In the homograft group there was a significant improvement in EF (p<0.05), and the exercise PG was significantly less. Both groups showed improved LV hypertrophy and correlation between V1S >24 mm criteria and LVMI measurements. Postoperative LDH levels in the homograft group were significantly lower than preoperative levels (p<0.05); the intergroup difference was also significant (p<0.001). CONCLUSIONS: Our data suggest that homografts, as compared to mechanical prostheses, provide significantly better hemodynamics in the aortic position. PMID- 10678376 TI - Wound healing in the mitral valve. AB - BACKGROUND AND AIMS OF THE STUDY: Limited information is available on the healing of wounds in the mitral valve. In the present study, this process has been evaluated in young sheep. METHODS: Histologic, histochemical and transmission electron microscopic studies were made of the healing of wounds produced surgically under cardiopulmonary bypass, in the mitral valve (a 1 cm-long radially oriented incision at the free edge of the anterior leaflet) of young sheep (n = 11). RESULTS: All animals developed severe mitral regurgitation, as demonstrated by hemodynamic, ultrasound and ventriculographic studies. At four to five weeks, the edges of the incision were only partially covered by endothelium and had small deposits of fibrin. Two types of granulation tissue were associated with wound healing. The first covered the cut edge and consisted of myxoid tissue that extended from the valvular spongiosa and fibrosa; the second type consisted of spindle-shaped cells arranged parallel to the surface and surrounded by a slightly myxoid stroma. This granulation tissue resembled that caused by reactive fibrosis in regurgitant mitral valves. The connective tissue cells in granulation tissue developed increasing amounts of actin-like filaments as a function of time after operation. This differentiation resulted in the development of myofibroblasts. In contrast to the avascular nature of normal mitral leaflets, capillaries and arterioles first appeared in the middle portion of the leaflet at eight weeks, and at the proximal edges of the wounds at 12 weeks. CONCLUSION: The healing of mitral valvular wounds is a slow process that requires between eight and 12 weeks for the formation of a dense collagenous scar at the edges and complete restoration of the endothelial lining. Progression of healing is associated with the phenotypic modulation of connective tissue cells from fibroblasts to myofibroblasts and neovascularization of the leaflets. PMID- 10678377 TI - Ischemic mitral valve repair surgery. AB - BACKGROUND AND AIM OF THE STUDY: The management of concomitant moderate or severe ischemic mitral regurgitation in the presence of ischemic heart disease is important for long-term prognosis. Mitral repair by either a suture or ring annuloplasty method has been advocated, although clear superiority of either method has not been established. METHODS: Combined coronary artery bypass and mitral valve surgery for ischemic mitral incompetence was performed on 68 consecutive patients between January 1996 and December 1998. The outcome in 63 of these patients (35 females, 28 males) who underwent mitral valve repair was reviewed. RESULTS: Average patient age was 61+/-9.4 years (range: 39-81 years). Average left ventricular ejection fraction (LVEF) was 42.1%; a suture annuloplasty was used in 84% and a ring in 16%. The average number of distal anastomoses was 3.9+/-1.1 (range: 1-6) and aortic cross-clamp time was 131+/-35 min (range: 58-238 min). Operative mortality rate (<30 days or in-hospital) was 12.7% and only requirement for intra-aortic balloon pumping either before or during surgery (21%) was predictive (p<0.05). On discharge, 98.2% of patients were in NYHA class I or II. Follow up (range: 1-35 months) was complete in 95% of cases. Moderate mitral regurgitation on discharge occurred in nine patients and was not related to the type of annuloplasty. Predictive risk factors were preoperative severe mitral regurgitation (p<0.04), poor LVEF (p = 0.05), and was predictive of deterioration of NYHA class (p<0.02), progression of regurgitation (p<0.05), and poor outcome (p<0.01). Poor outcome was also related to surgeon's experience. Structural valvular deterioration occurred in 21.8% of operative survivors, and there was one reoperation and four late deaths. The survival rate (including operative deaths) at 35 months was 68.3 +/- 13.1%, and event-free survival rate (no mortality, reoperation or angina) 65.2+/-6.2%. CONCLUSIONS: The type of annuloplasty used did not influence outcome. The risk of structural mitral valve dysfunction on follow up was related to severe preoperative mitral regurgitation, poor LVEF, surgeon's experience, and was predictive of poor outcome. PMID- 10678378 TI - Surgical repair of the prolapsing anterior leaflet in degenerative mitral valve disease. AB - BACKGROUND AND AIM OF THE STUDY: Repair of the prolapsing anterior leaflet (AML) in degenerative mitral valve disease is more demanding than that of the posterior leaflet. We reviewed our experience in the past eight years, to examine the safety, efficacy and stability of various repair artifices. METHODS: Between January 1989 and December 1997, 102 patients (mean age 64 years; range: 26-86 years) with mitral regurgitation (MR) due to prolapse of the anterior or both mitral leaflets underwent mitral valve repair. Sixty-six patients were in NYHA class > or =III, and 94 had MR grade >II. Acute endocarditis was present in 12 patients and Barlow disease in 16. Surgical techniques consisted of chordal shortening (n = 36), chordal transposition (n = 16), papillary muscle shortening or plication (n = 10), flip-over (n = 20) and artificial chordae implantation (n = 20). RESULTS: There was no early mortality; one patient required early mitral valve replacement (MVR) for late-appearing systolic anterior motion, and one patient benefited from a successful re-repair on day 8 for partial posterior leaflet desinsertion. Mean follow up was 30 months (range: 3-92 months); there were four late deaths (two valve-related cerebrovascular accidents); two patients required re-repair (one after three months for prosthetic ring thrombosis, and one after 10 months for rupture of shortened chordae (corrected by flip-over)). Five patients had MVR between four and 32 months later: one for mitral stenosis due to posterior leaflet calcification, and four for recurrent MR due to the rupture of shortened chordae (n = 3) or plicated papillary muscle (n = 1). One patient suffered bacterial endocarditis which was treated medically. Of the 92 remaining patients with valve repair, 81 are currently asymptomatic, five are in NYHA class II and four in class III. Transesophageal echocardiographic restudy (n = 76) at a mean of 30 months after surgery revealed no MR in 68 patients, and MR of grade or =70 years and with small aortic root diameters (< or =23 mm). PMID- 10678384 TI - First clinical experience with a mechanical valve with silver coating. AB - BACKGROUND AND AIM OF THE STUDY: The interface between the annulus and sewing cuff is the infectious center of prosthetic valve endocarditis (PVE). To decrease the incidence of PVE, the sewing cuff of the St. Jude Medical (SJM) mechanical heart valve was permanently coated with elemental silver (Silzone coating). In vitro data have supported the antimicrobial efficacy of this coating. METHODS: To study any adverse effects of the silver coating in humans, serum silver levels were determined (by graphite furnace atomic absorption spectrometry) before (baseline) and at five intervals after operation: day 1, day 3, discharge, one month, and two months. Between January and August 1997, 38 patients (71% males) underwent surgical implant of a SJM Masters Series valve with Silzone coating for the aortic valve (n = 29), mitral valve (n = 6), or both valves (n = 3). Five patients (13%) underwent concomitant procedures. Two patients (5%) presented with native active valve endocarditis. RESULTS: There was no hospital mortality or valve-related hospital morbidity. Blood silver concentrations peaked shortly after surgery and then decreased during the postoperative period. Average levels were consistently below 4 parts per billion (ppb). Levels below 10 ppb are considered normal. Follow up was 95% complete. There were no recurrent or new cases of endocarditis. CONCLUSION: These clinical data indicate that the SJM Masters Series valve with Silzone coating performs well. No adverse effects of the silver coating could be detected, and there were no valve-related complications. PMID- 10678385 TI - Preliminary experience with Silzone-coated St. Jude medical valves in acute infective endocarditis. AB - BACKGROUND AND AIM OF THE STUDY: The rate of recurrent postoperative endocarditis after valve replacement in early-stage acute infective endocarditis is extremely high. Metallic silver coating of the sewing ring may improve the short- and long term outcome after valve implantation. This report details our experience with the St. Jude Medical Silzone prosthesis in early surgical treatment of acute infective endocarditis. METHODS: Ten patients (mean age 66.4 years) referred for native valve or prosthetic valve endocarditis were operated on between April 1998 and June 1999. The microorganisms responsible for the acute infection were Staphylococcus (n = 1), Streptococcus (n = 1) and Pseudomonas aeruginosa (n = 1); blood cultures remained negative in two cases. The indication for surgical treatment was related to hemodynamic condition (n = 5), a major cerebral event (stroke; n = 1), annulus abscess (n = 1), and echocardiographic evidence of large cuspal vegetations (n = 3). All patients had received preoperative intravenous antibiotics (mean 7.8 days). Four mitral, five aortic valve replacements, and one double mitral-aortic valve replacement, were performed after extensive debridement of the infected and necrotic tissues. Mean duration of postoperative antibiotic treatment was 32.3 days. Postoperative follow up (mean 6 months; range: 2-14.2 months) was 100% complete, and included prospective repeated transthoracic echocardiography at one week, and one, six and 12 months postoperatively. RESULTS: One patient died early in the immediate postoperative period from pneumonia and major hypoxemia. All other patients are symptom-free, without evidence of recurrent infection and perivalvular leak. CONCLUSION: Although these early results with the St. Jude Medical Silzone prosthesis require confirmation by more extensive studies, they infer that silver coating of the sewing ring may dramatically improve management of patients with active endocarditis. PMID- 10678386 TI - In vivo efficacy of silver-coated fabric against Staphylococcus epidermidis. AB - BACKGROUND AND AIM OF THE STUDY: Prosthetic valve endocarditis (PVE) is a serious complication of heart valve replacement. The use of silver-coated polyester fabric in sewing cuff fabrication is intended to reduce the incidence of PVE. In this study, three pathogenic strains of Staphylococcus epidermidis (two PVE and one peripheral vasculature pathogen) were used to evaluate infection of silver coated and uncoated fabric. METHODS: Fabric was inoculated by preincubation for 30 min in bacterial suspensions containing 10(4), 10(5), 10(6), 10(7) or 10(8) CFU/ml, and implanted subdermally in mice for up to seven days. Bacteria adherent to fabric implanted for zero, one, three, or seven days were enumerated by sonicating the fabric and plating serial dilutions of the resulting suspension. Percent infection was assessed by implanting samples, subdermally, for seven days, then incubating explanted samples in growth media for three days and calculating the percent of fabric showing bacterial growth, for each concentration of inoculum. A logistic regression model was used to estimate curves relating percent infection to log concentration of the bacterial inoculum. These curves were used to estimate the concentration of inoculum required to produce 50% infection (ID50) for the three strains of S. epidermidis, for silver coated and uncoated fabric. RESULTS: Direct enumeration showed no difference in bacteria adherent to silver-coated and uncoated fabric, and no bacteria present in either fabric type in samples implanted for seven days. Nevertheless, incubation of those samples in growth media showed that many of the samples were infected. The calculated ID50 was significantly lower for silver-coated fabric than for uncoated fabric, for all three strains of S. epidermidis tested. CONCLUSION: Silver-coated fabric increases resistance to infection by S. epidermidis in this direct-contamination, mouse subdermal model. PMID- 10678387 TI - Examination of hemolytic potential with the On-X(R) prosthetic heart valve. AB - BACKGROUND AND AIM OF THE STUDY: Mechanical valves are known to produce chronic, subclinical hemolysis in most patients. Generally, haptoglobin is reduced to below normal in most patients, while lactate dehydrogenase (LDH) is increased to as much as 200% above the upper normal, sometimes resulting in anemia. The study was designed to investigate the clinical hemolysis of the On-X(R) prosthetic heart valve in a multicenter experience with a standard protocol and a single clinical laboratory. METHODS: Between September 1996 and August 1998, 248 patients underwent isolated valve replacement at 10 European centers. Blood samples were collected from these preoperatively and at 3-6 months and one year postoperatively. All samples were analyzed at a central laboratory, thus assuring poolability of the data. In total, 151 patients were tested at 3-6 months, and 62 at one year. Blood parameters measured were LDH, haptoglobin, hematocrit, total hemoglobin, red cell count and reticulocyte count. Paired analysis was used to compare preoperative baseline values with 3-6-month and one-year values. Data were analyzed with regard to both valve position and size. RESULTS: At 3-6 months and one year after surgery, average values for hematocrit, hemoglobin, red cell count and reticulocyte count were all near the center of the normal range, regardless of valve position or size. Statistically significant increases in red cell count and decreases in reticulocyte count occurred after both aortic valve replacement (AVR) and mitral valve replacement (MVR). These changes were of no clinical importance, but indicate that anemia has not occurred in these patients. At 3-6 months, haptoglobin was reduced to below normal in 86% of both AVR and MVR patients; this also occurred after one year and was statistically significant. Postoperatively, the mean LDH value in AVR was 228 U/l (91% of upper normal, 250 U/l) at 3-6 months, and 246 U/l (98% of upper normal) at one year. In MVR, these LDH values were 271 U/l (108% of upper normal) and 265 U/l (106% of upper normal). CONCLUSIONS: These results indicate that the On-X valve causes lower levels of chronic hemolysis in the immediate postoperative period and up to one year after surgery, especially when compared with reports of LDH elevations up to 200% of upper normal. Hemolytic anemia has not occurred in this patient population. PMID- 10678388 TI - Repeated thrombolysis in multiple episodes of obstructive thrombosis in prosthetic heart valves: a report of three cases and review of the literature. AB - BACKGROUND AND AIM OF THE STUDY: Thrombolysis is an emerging method to open thrombosed prosthetic heart valves. However, its applicability and safety in multiple recurrent thrombotic episodes is unclear. METHODS: Among 16 patients with thrombosed prosthetic valves treated with thrombolysis during a 33-month period, three patients (one mitral and two tricuspid) experienced four episodes each, and these were treated with repeated thrombolytic therapy. Data on patient demographics, clinical presentation, diagnosis, treatment and outcome are presented. RESULTS: Thrombolysis was successful in 10/12 episodes (83%); there was delayed response in one episode (8%), and partial response in one episode (8%). There were no major complications. However, a fifth thrombotic episode occurred in two patients with tricuspid prostheses, mandating re-do surgery. CONCLUSION: Thrombolysis in re-thrombosed prosthetic heart valves is feasible, highly successful and safe, and may therefore be used judiciously in selected patients who could not, or would not, undergo redo surgery. A high recurrence rate in the tricuspid position may implicate earlier surgical intervention, which should be individualized. PMID- 10678389 TI - Phenotypic and functional characterization of interstitial cells from human heart valves, pericardium and skin. AB - BACKGROUND AND AIM OF THE STUDY: Human heart valve interstitial cells (ICs) have been understudied to date. The aim of this study was to determine whether valve ICs were uniquely different from pericardial ICs and skin fibroblasts by determining their phenotype and investigating their reactivity. METHODS: ICs were cultured from human heart valves (n = 13), pericardium (n = 4) and skin (n = 4). Cell phenotype was determined by immunofluorescence using a panel of antibodies against surface and cytoskeletal components, and intracellular calcium changes were evaluated by loading the cells with fura-2 acetoxymethyl ester. RESULTS: Skin fibroblasts expressed virtually no smooth muscle (SM) alpha-actin, whereas, 56.9+/-8.9% (mean +/- SEM) of ICs from valves and 21+/-7.6% of ICs from pericardium expressed SM alpha-actin. ICs from pericardium and skin fibroblasts always expressed a fibroblast surface antigen, whereas expression was variable on valve ICs (80+/-6.9%). All cells expressed prolyl 4-hydroxylase, beta-tubulin and vimentin, but not desmin. Transient increases in intracellular calcium were induced by vasoactive agents: skin fibroblasts were least responsive to all agents. CONCLUSIONS: ICs cultured from heart valves consist of a mixed population of specific cell types, many of which express SM alpha-actin and may be classified as myofibroblasts. The intracellular calcium responses to vasoactive agents indicated that a number of receptor signaling pathways existed. Evaluation of these may help to elucidate the role of myofibroblasts and other fibroblast phenotypes in valve function/dysfunction, as well as contribute to the development of tissue-engineered valves. PMID- 10678390 TI - The dilemma of accelerated bioprosthetic and polymeric valve testing. PMID- 10678391 TI - Fatigue analysis of clinical bioprosthetic heart valves manufactured using photooxidized bovine pericardium. AB - BACKGROUND AND AIM OF THE STUDY: Recent reports have given details of early clinical failures of bioprosthetic heart valves manufactured using dye-mediated photooxidized bovine pericardium. These failures were attributed to abrasion of the inflow surface of the leaflets against the cloth-covered inner face of the outer valve frame. These failures had not been detected during preclinical testing of such valves. The aim of this study was to determine if such failure modes could be replicated during fatigue testing of the photooxidized valves, and whether lining of the inflow face of the outer frame with pericardium could eliminate these failures. METHODS: The fatigue properties of six lined and six unlined PhotoFix(R) alpha valves was determined using two Rowan Ash fatigue testers which were cycled at 15 Hz for a maximum of 210 million cycles. The closing pressure within in each chamber was 110+/-10 mm Hg. Each valve was inspected every 40 million cycles for any signs of fatigue failure. All valves were tested to at least 210 million cycles. RESULTS: In all six lined PhotoFix alpha valves there was no evidence that wear, tear or abrasion of the inflow aspect of the leaflets adjacent to the inflow face of the outer frame had occurred. Only one unlined valve showed any signs of damage adjacent to the outer frame, as evidenced by loss of loose connective tissue, but this did not have the imprint of the cloth covering, which would have suggested actual wear. Effectively, after 200 million cycles, no fatigue failures of either the lined or the unlined PhotoFix alpha valves had occurred. Moreover, the leaflet tears that had been observed clinically were not replicated in this study. CONCLUSION: Previous studies have shown a good qualitative, quantitative association between fatigue failure in Rowan Ash accelerated fatigue testers, and clinical experience in glutaraldehyde-treated bovine pericardial valves. The accelerated fatigue tester did not provide an adequate model for the prediction of clinical failure for the photooxidized pericardial valves. PMID- 10678392 TI - Future directions in tissue heart valves: impact of recent insights from biology and pathology. PMID- 10678393 TI - Mitral valve replacement with a pulmonary autograft: initial experience. PMID- 10678394 TI - Sheep CD1 genes and proteins. AB - Interest in CD1 genes and proteins was initially stimulated by their close evolutionary and structural relationship to MHC class I molecules. The demonstration that CD1b and c molecules present novel non-peptide antigens to T cells and play a role in protection against mycobacterial infection then focused attention on the functional role of CD1 proteins. Sheep possess at least seven CD1 genes, including CD1B, D and E, which is the most complex genetic arrangement identified so far in any animal. OvCD1B consists of at least three distinct genes, with the probability of limited polymorphism and the existence of splice variants. Most anti-sheep CD1-specific monoclonal antibodies react with OvCDlb and phenotypic and immunochemical data suggests the existence of two variants. CD1D genes have been identified in all species studied, suggesting a conserved role for CDld proteins across mammalian species. Presumptive evidence for the existence of OvCDIE has been obtained by NH2-terminal sequencing of protein precipitated by the mAb 20.27 (SBU-T6). Confirmatory evidence from gene cloning experiments is currently being sought. Collectively, these factors make the sheep CD1 family a highly relevant model for investigating the in vivo role of CD1 molecules. In this survey, the properties of monoclonal antibodies specific for sheep CD1, the cellular distribution and physicochemical characteristics of sheep CD1 molecules and the current state of knowledge on sheep CD1 genetics are reviewed. PMID- 10678395 TI - Lymphocyte populations and adhesion molecule expression in bovine tonsils. AB - Bovine tonsils are mucosal-associated lymphoid tissue (MALT) located at the entry of the pharynx where both inhaled and ingested antigens can induce an immune response. This study was conducted to determine the lymphocyte populations and adhesion molecule expression in the palatine tonsil (PT) and pharyngeal tonsil (PhT) of adult cattle and compare them with typical MALT (discrete Peyer's patches, PP) and a peripheral lymph node (parotid lymph node, PLN). The distribution of various lymphocyte subsets was determined in situ by immunofluorescence, and their proportions were determined by multicolor flow cytometry. The tonsils were similar to PP in the proportions of B- and T-cells (25-32% T-cells, 39-45% B-cells), and T cell subpopulations (CD4, CD8, and gammadelta). The PP contained the highest proportion of memory T-helper cells with beta7 integrin (30.3%+/-5.4), the tonsils intermediate (PT: 19.8%+/-4.4 and PhT: 19.7%+/-4.9), and the PLN had the lowest proportion (15.4%+/-3.1). The opposite relationship was observed with CD62L on naive T- helper cells as PP had the lowest proportion (14.2%+/-6.4), the tonsils intermediate (PT: 17.4%+/-2.5 and PhT: 24.3%+/-7.3), and the PLN the highest proportion (45.3%+/-6.5). MAdCAM-1 was highly expressed in the high endothelial venules (HEV) of PP, with variable and weak expression in the tonsils and PLN. PNAd, on the other hand, was highly expressed in HEV of tonsils and PLN, and weakly expressed in the PP. These results indicate that the bovine tonsils share characteristics with both PP and PLN. The alpha4beta7/MadCAM-land CD62L/PNAd interaction may be involved in lymphocyte migration to the tonsils, but it is likely that other adhesion molecules participate as well. Similarities between the human and bovine tonsils suggest that cattle may provide a good model to study the role of the tonsil in the respiratory immune response. PMID- 10678396 TI - Molecular cloning, phylogenetic analysis and expression of beluga whale (Delphinapterus leucas) interleukin 6. AB - Interleukin 6 (IL-6) is a cytokine produced primarily by the monocytes/macrophages with regulatory effects in hematopoiesis, acute phase response, and multiple aspects of the immune response. IL-6 exerts its activity through its binding to specific high affinity receptors at the surface of target cells. As yet, no molecular data have been reported for the beluga whale IL-6. In this study, we cloned and determined the entire beluga whale IL-6-encoding cDNA sequence by reverse transcription-polymerase chain reaction (RT-PCR) sequencing, and analysed its genetic relationship with those from several mammalian species including human, rodent, ruminant, carnivore and other marine species. The identity levels of beluga whale IL-6 nucleic and deduced amino acid sequences with those from these mammalian species ranged from 62.3 to 97.3%, and 42.9 to 95.6%, respectively. Phylogenetic analysis based on amino acid sequences showed that the beluga whale IL-6 was most closely related to that of the killer whale. Thereafter, beluga whale IL-6-encoding sequence was successfully expressed in Escherichia coli by using the pTHIOHisA expression vector for the production of a recombinant fusion protein. The immunogenicity of the recombinant fusion protein was then confirmed as determined by the production of a beluga whale IL-6 specific rabbit antiserum. PMID- 10678397 TI - Equine bullous pemphigoid IgG autoantibodies target linear epitopes in the NC16A ectodomain of collagen XVII (BP180, BPAG2). AB - Bullous pemphigoid (BP) is an autoimmune subepithelial blistering dermatosis of humans, dogs, cats and pigs. It is characterized by skin-fixed and circulating IgG autoantibodies that target one or both BP antigens. An immunological homologue of BP in humans was diagnosed in two horses with cutaneous and mucosal ulcerations as well as microscopic subepithelial vesiculation. Immunological investigations revealed similar findings for both the horses. Direct immunofluorescence demonstrated the presence of IgG deposited linearly at the dermoepidermal junction in mucosal and skin biopsy specimens. Indirect immunofluorescence testing confirmed the existence of circulating basement membrane-specific IgG autoantibodies. Using intact and salt-split epithelial substrates, serum IgG were shown to target antigens situated not only at the basal, but also at the lateral and apical aspects of stratum basale keratinocytes. Immunoblotting and ELISA corroborated that the IgG from affected horses, but not those from normal controls, exhibited high immunoreactivity against the NC16A extracellular domain of type XVII collagen (BPAG2, BP180). Equine BP could be proposed, therefore, as another spontaneous model of this most common basement membrane autoimmune dermatosis of humans. PMID- 10678398 TI - Identification of two allelic IgG1 C(H) coding regions (Cgamma1) of cat. AB - Two types of cDNA encoding IgG1 heavy chain (gamma1) were isolated from a single domestic short-hair cat. Sequence analysis indicated a higher level of similarity of these Cgamma1 sequences to human Cgamma1 sequence (76.9 and 77.0%) than to mouse sequence (70.0 and 69.7%) at the nucleotide level. Predicted primary structures of both the feline Cgamma1 genes, designated as Cgamma1a and Cgamma1b, were similar to that of human Cgamma1 gene, for instance, as to the size of constant domains, the presence of six conserved cysteine residues involved in formation of the domain structure, and the location of a conserved N-linked glycosylation site. Sequence comparison between the two alleles showed that 7 out of 10 nucleotide differences were within the C(H)3 domain coding region, all leading to nonsynonymous changes in amino acid residues. Partial sequence analysis of genomic clones showed three nucleotide substitutions between the two Cgamma1 alleles in the intron between the CH2 and C(H)3 domain coding regions. In 12 domestic short-hair cats used in this study, the frequency of Cgamma1a allele (62.5%) was higher than that of the Cgamma1b allele (37.5%). PMID- 10678399 TI - Production and characterisation of two monoclonal antibodies recognising equine IgG Fc receptors. AB - Despite the importance of IgG Fc receptors in the regulation of various immunological mechanisms, these receptors have not been well characterised in the domesticated animals including equines. This paper describes the production of two monoclonal antibodies (CVS 59 and CVS 61) that recognise equine IgG Fc receptors. Fusions were conducted using BALB/c mice hyperimmunised with equine peripheral blood mononuclear cells. Hybridoma supernatants were screened on the basis of their ability to inhibit the rosetting of equine antibody coated sheep erythrocytes with equine peripheral blood mononuclear cells. The mAbs were then characterised for cellular distribution of the antigen by flow cytometry. Using immunoprecipitation, it was shown that both under reducing and non-reducing conditions, CVS 59 and CVS 61 precipitated a 80 and 40 kD protein, respectively. From the functional study coupled with cellular distribution and molecular weight of the antigen recognised by these mAbs, it would appear that CVS 59 and CVS 61 recognise an epitope on equine equivalent of human and murine FcgammaRIII and FcgammaRII, respectively. PMID- 10678400 TI - Induction of aggregation in porcine lymphoid cells by antibodies to CD46. AB - CD46 is a major transmembrane glycoprotein that belongs to the regulator of complement activation (RCA) family. Recently, mAbs to human CD46 were shown to suppress IL-12 production. Here, we describe that mAbs against different porcine CD46 epitopes induced a marked adhesion of normal lymphocytes. Addition of low amounts of antibody to freshly isolated lymphocytes or thymocytes resulted in the clustering of the cells. Cross-linking of CD46 molecules seems essential since Fab fragments failed to induce aggregation. This aggregation was dependent on active cell metabolism and on the presence of divalent cations and required a functional cytoskeleton. It was not inhibited by antibodies to CD18, CD29, CD2, CD11a and CD11b. Staurosporine, an inhibitor of protein kinases, partially blocked the aggregation. This finding is indicative of a role of protein kinases in the transduction of the signal generated by CD46 engagement. PMID- 10678401 TI - Phenotypical characterization of T and B cell areas in lymphoid tissues of dogs with spontaneous distemper. AB - CD3, CD4, CD5, and CD8 antigen expression of T cells and IgG expression of B cells and canine distemper virus (CDV) antigen distribution were immunohistochemically examined in lymphoid tissues (lymph node, spleen, thymus, and tonsil) of control dogs and animals with spontaneous canine distemper. In addition, CNS tissue of all animals was studied for neuropathological changes and CDV antigen distribution. Based on the degree of depletion distemper dogs were classified into two groups. Group I represented animals with moderate to marked lymphoid depletion, while group II dogs displayed mild or no depletion. CDV antigen was mainly found in lymphocytes and macrophages of group I dogs, whereas CDV expression was most prominent in dendritic cells of group II animals. In group I dogs, a marked loss of CD3, CD4, CD5, CD8, and IgG expression was noticed, hereby loss of CD4+ cells was more prominent than depletion of CD8+ cells. In the lymphoid tissues of group II animals, a significant increase in the number of T and B cells was observed compared to group I dogs. The number of CD3+, CD4+, and CD8+ cells in group II dogs was similar to the findings in controls, however, CD5 and IgG expression was mildly reduced in T and B cell areas, respectively. Additionally, in groups I and II dogs, CD3+ and CD5- T cells were detected in T cell areas. Whether this cell population represents a cell type with autoimmune reactive potential remains to be determined. Surprisingly in group II animals, viral antigen was found predominantly in dendritic cells indicating a change in the cell tropism of CDV during chronic infection and a possible mechanism of viral persistence. The two patterns of lymphoid depletions correlated to two different types of canine distemper encephalitis (CDE). Group I dogs displayed acute non-inflammatory CDE, whereas group II dogs suffered from chronic inflammatory demyelinating CDE, indicating a pathogenic relationship between lymphocytic depletion and inflammatory brain lesions in distemper. PMID- 10678402 TI - The possible role of osteoclastogenic oral bacterial products in etiology of Paget's disease. PMID- 10678403 TI - Normative data for iliac bone histomorphometry in growing children. AB - Many insights into normal and pathologic bone development can only be gained by bone histomorphometry. However, the use of this technique in pediatrics has so far been hampered by the lack of reference data. Therefore, we obtained transfixing iliac bone samples from 58 individuals between 1.5 and 22.9 years of age (25 male; tetracycline labeling performed in 48 subjects), who underwent surgery for reasons independent of abnormalities in bone development and metabolism. The results of histomorphometric analyses of cancellous parameters and cortical width are presented as means and standard deviations, as well as medians and ranges in five age groups. In addition, the original data are available from the authors. There were significant age-dependent increases in both cortical width and cancellous bone volume, the latter being due to an increase in trabecular thickness. Osteoid thickness did not vary significantly with age. Bone surface-based indicators of bone formation showed an age-dependent decline, reflecting similar changes in activation frequency. Mineral apposition rate decreased continuously with age. Parameters of bone resorption did not vary significantly between age groups. Paired biopsies from adjacent sites, obtained in eight subjects, were used to examine the reproducibility of histomorphometric parameters in children. The lowest coefficients of variation (<10%) were found for structural measures, as well as mineral apposition rate and wall thickness. The highest variability was found for cellular parameters. The availability of reference material will greatly facilitate the use of histomorphometry in pediatrics. PMID- 10678404 TI - No major effect of estrogen receptor gene polymorphisms on bone mineral density or bone loss in postmenopausal Danish women. AB - The polymorphisms of the estrogen receptor (ER) gene defined by the restriction enodonucleases PvuII and XbaI have recently been reported to be associated with bone mineral density (BMD) in postmenopausal women. To investigate the possible relation of the PvuII and XbaI restriction fragment-length polymorphisms of the ER gene with BMD in Danish postmenopausal women, two studies were undertaken: 1) a cross-sectional study of 499 postmenopausal women, where the ER genotypes and alleles were related to BMD of the hip, spine, and lower forearm; and 2) a longitudinal study of 101 postmenopausal women followed up for 18 years. In the latter study, late postmenopausal bone loss in the hip and spine was determined over a period of 6 years in women (mean age of 63 to 69 years), and long-term postmenopausal bone loss in the lower forearm was determined over a period of 18 years in women (mean age of 51 to 69 years). Genotyping was performed through the restriction cleavage of polymerase chain reaction-amplified genomic DNA with the two restriction enzymes, PvuII and XbaI. Restriction fragment-length polymorphisms were represented as P or p (PvuII) and X or x (XbaI), with the lower case letters signifying the presence of the restriction site. The frequencies of the ER genotypes were similar to previously published genotype frequencies in Caucasian and Asian populations. No significant effect of the ER genotypes or alleles on BMD was found at any site, nor was there a relation between ER genotypes and the rate of bone loss either in the hip and spine over 6 years, or in the lower forearm over 18 years. In conclusion, we could not demonstrate any major effect of the ER gene polymorphisms on BMD or rate of bone loss in healthy postmenopausal Danish women. PMID- 10678405 TI - Cellular distribution of estrogen receptor beta in neonatal rat bone. AB - Estrogens affect bone metabolism, and ovariectomy of rats results in marked bone loss caused by stimulation of osteoclastic bone resorption. Estrogen receptors (ER) have been demonstrated in osteoblasts and bone marrow stromal cells, but their presence in osteoclasts is controversial. Until recently, only one type of ER (now renamed ERalpha) had been identified. After the discovery of a novel ER subtype (ERbeta), it became necessary to re-investigate the ER expression in human and rodent bone. In the present study we examined the expression of ER mRNA in neonatal rat bone. Expression of ER alpha and beta mRNA (RT-PCR) was evident in femurs of 3-week-old male and female rats. In situ hybridization histochemistry of femural bones with digoxigenin labelled riboprobes, as well as radioactively labeled riboprobes, revealed that ERbeta mRNA was predominantly expressed in osteoblasts covering the metaphyseal bone trabecular surface. The presence of ERbeta mRNA in osteoblasts of rat bone suggests that ERbeta is involved in the mechanism of action of estrogens in bone. PMID- 10678406 TI - Cross-sectional assessment of age-related bone loss in men: the MINOS study. AB - There are few data on osteoporosis in men, but cross-sectional studies have shown that age-related bone loss in men is of lower magnitude than in women. To elucidate some controversies related partially to methodological aspects, we measured bone mineral density (BMD) by dual-energy X-ray absorptiometry (DEXA) at various skeletal sites (spine, hip, and whole body using a Hologic QDR-1500 device; forearm using an Osteometer DTX 100 device) in a large cohort of 1040 men, aged 19-85 years. The final investigation was performed on 934 men, aged 19 85 years, after exclusion of 106 men with disease or treatment known to affect bone metabolism. Peak BMD was achieved at 25 and 29 years at the lumbar spine and hip, respectively, but only at 40 and 37 years at the distal forearm and whole body, respectively. The magnitude of bone loss between peak bone mass and 80 years of age was linear at most sites and averaged 13%-18%; that is, SD of 1.1 1.8 from peak BMD, except for Ward's triangle, which showed a marked bone loss of 43% (i.e., 2.5 SD), and for the lumbar spine. In the entire cohort, increase of the average lumbar spine BMD after the age of 55 years was related to the development of osteoarthritis, because, in men without severe arthritis, lumbar spine BMD continued to decrease. Height-adjusted partial correlations indicate that both the mineral content and the area of long bones of the limbs increased with age up to 50 years, followed by a significant decrease of BMD without change of bone surface. SD of mean BMD increased significantly with age at most skeletal sites. In summary, age-related change of BMD varied according to skeletal site in men with peak bone mass achieved earlier at sites rich in trabecular bone than at those rich in cortical bone. Bone loss varied according to skeletal site from 14% to 43%. The variability of BMD increased with age, which may reflect interindividual variability of age-related bone loss. PMID- 10678407 TI - Cross-sectional and longitudinal assessment of pre- and postmenopausal bone loss with a portable forearm X-ray device: the Ofely study. AB - The aim of our study was to describe cross-sectional and longitudinal bone mineral decrease in pre-, peri-, and postmenopausal healthy women using a monoenergy X-ray densitometer specifically designed for forearm assessement. Measurements were performed on the most distal part of the radius (ultradistal, 55% of trabecular bone and 45% of trabecular bone), and on the distal part (distal, 13% of trabecular bone and 87% of cortical bone). A specific trabecular rich region of interest (nROI) comprising two trapezoids regions of interest located proximally to the endplates of the radius and ulna was also investigated. From a large prospective study (OFELY study), 455 women were measured once a year for 2 years (three measurements). The proportion of postmenopausal women classified as having osteoporosis (i.e., a T score <-2.5) was 33% for the distal region, 44% for the ultradistal region, and 45% for the nROI. No significant bone mineral decrease was found over the 2-year period in premenopausal women (n = 138). In perimenopausal (n = 48) women, a bone loss of 1% was found at the distal site. In the 269 postmenopausal women, a significant decrease was observed at all sites, ranging from 2.14% for the nROI to 2.68% for the ultradistal part. Bone loss was greater in the first 5 years after menopause in trabecular sites and decreased thereafter. For the distal site, bone loss remained stable during the postmenopausal period. We conclude that this small and portable forearm densitometer is suitable for the diagnosis of osteoporosis, and provides information on the rate of bone loss in peri- and postmenopausal women in trabecular and cortical compartments of bone. PMID- 10678408 TI - Cytokine RNA levels in transiliac bone biopsies from healthy early postmenopausal women. AB - The cytokines interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF alpha), and IL-6 induce osteoclast formation and may contribute to the development of postmenopausal osteoporosis. Cross-sectional studies have suggested that both IL-1 and IL-1ra secretion increase on estrogen withdrawal, and that postmenopausal osteoporosis is associated with an inadequate increase in monocyte IL-1ra secretion with age. We measured cytokine mRNA (IL-1beta, IL-1ra, IL-6, and TNF-alpha) directly in bone biopsies from early postmenopausal women to determine if a lower compensatory increase in IL-1ra mRNA could be demonstrated in women with rapid bone loss after the menopause. Biopsies were obtained from 23 early postmenopausal women (mean age 53.9 years) who participated in a randomized study of hormone replacement therapy (HRT) and risk factors for osteoporosis. Bone mineral density was assessed by duel energy X-ray absorptiometry at 0, 1, 2, and 5 years. Women in the control group were recruited to the biopsy study based on their observed rate of bone loss (upper or lower tertile). Consent was also obtained from 11 participants receiving HRT. Biopsies were taken at 2 years, frozen in nitrogen, and homogenized. Cytokine mRNA was measured by competitive reverse transcriptase polymerase chain reaction. The IL-1ra/IL-1beta mRNA slope for the slow-loss group was steeper (deltaF = 23.3, p < 0.01) than that observed in the fast-loss group, indicating that slower bone loss was associated with higher IL-1ra mRNA levels relative to IL-1beta. During HRT, the IL-1beta mRNA level was inversely correlated with serum estradiol (log r2 = 0.77, p < 0.01), and women with a serum estradiol below 200 pmol/L during HRT had IL-1beta, mRNA levels identical to the control group. In contrast, IL-1ra mRNA was independent of serum estradiol. Histomorphometric analysis revealed weak correlations between IL-1beta mRNA and activation frequency (r2 = 0.26, p = 0.06) and between IL-1ra and volume referent bone resorption rate (r2 = 0.19, p = 0.11). TNF-alpha was not associated with the bone loss rates or with serum estradiol, and only three samples were positive for IL-6 mRNA. The findings support the hypothesis that IL 1beta production within bone increases with declining estrogen levels, and that an increase in II-1ra protects against accelerated bone loss. PMID- 10678409 TI - In vivo diffuse damage in human vertebral trabecular bone. AB - Accumulation of microdamage in vivo may lead to loss of bone quality. Until recently, linear microcracks were the only known form of in vivo microdamage, but through the use of confocal microscopy an additional level of damage (diffuse damage) has been identified. In this study, in vivo diffuse damage was characterized and quantified in human vertebral trabecular bone as a function of tissue morphology, age, race, gender, and previously quantified in vivo linear microcracks. Presence of diffuse damage in human vertebral tissue was confirmed and validated by simultaneous use of polarized, ultraviolet, and laser confocal microscopy. Diffuse damage was found to occur preferentially within trabecular packets rather than in interstitial bone (p < 0.05). It was consistently higher in men compared with women (p < 0.05), but was not different by race or age group. Diffuse damage did not correlate with linear microcracks, but both exhibited the same probability distribution in which the percentage of individuals having a particular amount of damage decreased exponentially as damage content increased. These findings suggest that diffuse damage accumulation and repair are governed by the same biological phenomena as microcracks, but diffuse damage contributes independently to the microdamage content of bone. PMID- 10678410 TI - Does the mechanical milieu associated with high-speed running lead to adaptive changes in diaphyseal growing bone? AB - Exercise during growth can be important for attaining optimal bone mass. High intensity long-duration protocols, however, can have detrimental effects on immature bone morphology and mechanics. The underlying mechanisms are poorly understood. Here, we quantified the mechanical environment of the middiaphyseal rooster tarsometatarsus during high-speed running and examined whether short bouts of this exercise-related mechanical milieu can induce positive changes in cortical bone morphology, mechanics, and mineral ash content. At 9 weeks of age, roosters were assigned to controls (n = 9) and runners (n = 8). Treadmill running was applied in loading sessions of 5 min, three times per day (approximately 2600 cycles/day) for 8 weeks. Both controls and runners received double-fluorochrome labels during weeks 3 and 8 of the protocol. Middiaphyseal distributions of tarsometatarsal longitudinal normal strain, strain rate, and strain gradients engendered by walking and running were determined via in vivo strain gauges. Compared with walking, running elevated mean peak strain magnitude by 19%, peak strain rates by 136%, and peak strain gradients by approximately 18%. After 8 weeks of running, middiaphyseal areal and mechanical properties and normalized ash weight were no different between runners and controls. Transient and focal reductions in periosteal mineral apposition rates occurred during the exercise protocol. Our current data suggest that reducing the number of loading cycles can mitigate the adverse response previously observed in this model with long duration running. This study also supports the tenet that the exercise-generated mechanical milieu must differ substantially from the habitual milieu to induce significant adaptations. PMID- 10678411 TI - Effects of recombinant human fibroblast growth factor-2 on osteogenic cell populations during orthopic osteogenesis in vivo. AB - The osteogenic effects of fibroblast growth factor-2 (FGF-2) in vivo on different cell populations of the osteoblastic cell lineage have not been fully elucidated. In this study, the efficacy of recombinant human fibroblast growth factor-2 (rhFGF-2) to stimulate orthopic bone formation in transosseous rat mandibular defects, with different cell populations allowed access to the defects, was investigated with the aim to further decipher FGF-2 effects. Three different doses of rhFGF-2 (10 ng, 100 ng, and 1 microg) were delivered in an absorbable collagen sponge carrier, whereas some defects were implanted with the carrier only, and some were left untreated. Barrier membranes, made of microporous expanded polytetrafluoroethylene, were simultaneously placed over half the number of defects in each treatment group, thus forcing osteogenic cells to be derived from intraosseous sources. Evaluation was made by light microscopy and computerized image analysis after 12 and 24 of days healing. Whereas no general stimulatory effect could be ascertained at 12 days, higher rhFGF-2 doses decreased bone formation by both intraosseously and periosteally derived cells. At 24 days, a clear, although rather limited, stimulatory effect on osteogenesis was observed, but again a decrease was observed with the 1 microg dose. At both observation periods, an increased number of osteocytes was found in the newly formed bone at sites treated with the lower rhFGF-2 doses, whereas the high-dose rhFGF-2 resulted in a return to control levels, irrespective of whether cells were intraosseously derived or from the periosteum also. Based on differential analysis of bone healing by cells from different sources as well as on bone cellularity, the results suggest that rhFGF-2 in vivo exerts a stimulatory effect on proliferation of committed osteoblastic cells. This effect is biphasic, in that higher doses are without effect or may even be inhibitory. No inductive effect on osteoblast recruitment could be found. These effects differ from those of, for instance, BMP-2 and TGF-beta1. PMID- 10678412 TI - Bone stiffness predicts strength similarly for human vertebral cancellous bone in compression and for cortical bone in tension. AB - The yield strength and ultimate strength of cortical and cancellous bone tissue are very highly correlated to bone stiffness. For samples of human vertebral cancellous bone in compression and for bovine cortical bone in tension, the coefficient of determination (r2) for regression between ultimate strength and stiffness was 0.89 and 0.92, and between yield strength and stiffness it was 0.94 and 0.93, respectively. The slope of the regression for human vertebral cancellous bone ultimate strength predicted by stiffness was not statistically different from similar regressions for cortical bone in tension in either a bovine sample or in published data from multiple species. We believe that the observed correlation results from the evolutionary need to build sufficiently strong bones using cells that are sensitive to deformation and that directly control bone stiffness, but not strength. The practical significance of this work is that an in vivo estimate of bone stiffness (e.g., from ultrasound measurement) may be a surrogate for bone strength. PMID- 10678413 TI - Daily intermittent decreases in serum levels of parathyroid hormone have an anabolic-like action on the bones of uremic rats with low-turnover bone and osteomalacia. AB - The calcium receptor agonist (calcimimetic) compound NPS R-568 causes rapid decreases in circulating levels of parathyroid hormone (PTH) in rats and humans. We hypothesized that daily intermittent decreases in serum PTH levels may have different effects on bone than do chronically sustained decreases. To test this hypothesis, we compared two NPS R-568 dosing regimens in rats with chronic renal insufficiency induced by two intravenous injections of adriamycin. Fourteen weeks after the second adriamycin injection, creatinine clearance was reduced by 52%, PTH levels were elevated approximately 2.5-fold, and serum 25(OH)D3 and 1,25(OH)2D3 levels were reduced substantially. Treatment by daily per os gavage, which decreased PTH levels intermittently, or continuous subcutaneous infusion, which resulted in a sustained suppression of serum PTH levels, then began for 8 weeks. Despite the hyperparathyroidism, the adriamycin-injected rats developed a low-turnover bone lesion with osteomalacia (fourfold increase in osteoid volume in the proximal tibial metaphysis) and osteopenia (67% decrease in cancellous bone volume and an 18% reduction in bone mineral density at the distal femur). Daily administered (but not infused) NPS R-568 significantly increased cancellous bone volume solely by normalizing trabecular thickness, and increased femoral bone mineral density by 14%. These results indicate that daily intermittent, but not sustained, decreases in PTH levels have an "anabolic-like" effect on bones with a low-turnover lesion in this animal model of chronic renal insufficiency. PMID- 10678414 TI - A murine model of inflammatory bone disease. AB - We have recently reported the identification of a new recessive mutation on murine chromosome 18 that results in tail kinks and deformity in the lower extremities of mice. Preliminary examination of the bones of these mice showed that there are abnormalities present that resembled chronic recurrent multifocal osteomyelitis. Accordingly, this new mutation was named "CMO." In this report, we describe the histology of bones in CMO mice, as well as the capacity of the bone marrow cells from these animals to form osteoclasts (OCLs). In addition, we tested conditioned media from non-adherent marrow cells and total marrow cells from CMO mice for their capacity to induce OCL formation in normal murine marrow cultures. These studies demonstrated that the bone disease in these animals is inflammatory in nature, and a soluble factor(s) that is not IL-1alpha, IL-6 or TNF-alpha is released by marrow cells from CMO animals and enhances OCL formation in normal murine marrow cultures. PMID- 10678415 TI - Relationships between bone protein and mineral in developing porcine long bone and calvaria. AB - Several proteins in the bone matrix have been implicated in the regulation of mineral crystal formation and growth. To investigate the relationships between these proteins and the mineral phase at various stages of mineral maturation, fetal porcine calvariae and long bones were fragmented and the particles (20 microm) separated by density gradient sedimentation into fractions of increasing density (1.8 to >2.2 g/cm3). Samples from each fraction were analyzed by X-ray diffraction to obtain the average crystal size/strain and chemical composition. Other samples were sequentially extracted, first with 4.0 mol/L guanidium hydrochloride (GuHCl) (G1), then with 0.5 mol/L ethylene-diamine tetraacetic acid (EDTA) (E), and again with 4.0 mol/L Gu-HCI (G2), for analysis of proteins in different tissue compartments. Based on the mineral density distribution and crystal size, fetal porcine bone protein content was determined for tissue residue and each extract and the protein composition analyzed by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE). Although the insoluble organic matrix decreased with mineral density the collagen and protein content remained fairly constant, representing approximately 10% of the tissue weight, except in the highest density fraction. Whereas the total extractable protein, representing predominantly noncollagenous proteins, did not show density-related differences, differences were observed for individual proteins on SDS-PAGE. Consistent with their presence in osteoid, the content of bone sialoprotein (BSP), tyrosine-rich acidic matrix protein (TRAMP), and a series of small proteins with cell attachment properties in the G1 extract decreased with mineral density, whereas TRAMP and BSP were increased in G2 extracts. Mineral-associated proteins, including alpha2HS-glycoprotein, BSP, osteopontin (OPN), and osteocalcin, increased with mineral density, whereas secreted protein acidic and rich in cysteine (SPARC)/osteonectin, and some minor proteins, appeared to decrease. Differences of individual proteins within and between the calvarial and long bones could be related to the role of these proteins in the formation and maturation of hydroxyapatite crystals. Collectively, these studies demonstrate mineral density-associated changes in protein composition that reflect a rapid maturation of mineral crystals in embryonic porcine bones. PMID- 10678416 TI - Criteria used to identify osteoclasts formed in vitro. PMID- 10678417 TI - Multidrug-resistant Salmonella Typhimurium DT104 in poultry. AB - Salmonella Typhimurium isolates from feed ingredients or poultry sources isolated during 1995 to 1997 from different geographical locations within Minnesota were examined for the presence of Salmonella Typhimurium definitive type 104 (DT104). Antibiotic susceptibility studies indicated that 15 of 50 isolates of Salmonella Typhimurium had an antibiotic resistance pattern (ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline) that is usually observed with multidrug-resistant Salmonella Typhimurium DT104. Of the 15 isolates showing the antibiotic resistance pattern, 8 isolates were phage type 104, 3 isolates were typed as phage type 104 complex, and the remaining 4 isolates belonged to phage types 193, 81, and 126. DT104 was recovered from both feed ingredients and poultry samples. Of the seven feed ingredient-associated Salmonella Typhimurium isolates, four were DT104, whereas only 7 of 43 poultry-associated Salmonella Typhimurium isolates were DT104. A repetitive sequence-based polymerase chain reaction (rep-PCR) of 50 isolates of Salmonella Typhimurium representing 13 phage types identified seven distinct fingerprint profiles. No correlation between phage type and rep-PCR type was noticed. Eleven Salmonella Typhimurium isolates belonging to DT104 and its complex were grouped into two closely related rep-PCR types. PMID- 10678418 TI - Survival of Pseudomonas fluorescens and Salmonella typhimurium after electron beam and gamma irradiation of refrigerated beef. AB - The radurization effects of gamma ray and electron beam irradiation at 1.5 and 3.0 kGy on beef steaks inoculated with Salmonella Typhimurium and Pseudomonas fluorescens were investigated during 8 days of storage at 5 degrees C. Total bacterial counts and numbers of Salmonella Typhimurium and P. fluorescens were analyzed at 2-day intervals. Total bacterial counts of samples irradiated by both gamma rays and electron beam were significantly (P < 0.05) reduced by 3.8 to 5.3 log CFU/g. Salmonella Typhimurium was not detectable during the experimental period. P. fluorescens counts of beef samples irradiated by gamma rays at both 1.5 and 3.0 kGy were not detected; however, P. fluorescens in samples irradiated by electron beam at 1.5 and 3.0 kGy was recovered after 2 days, and bacterial counts reached 7.8 and 6.9 log CFU/g, respectively. Both gamma ray and electron beam irradiation reduced total bacterial counts initially, possibly extending shelf life. Irradiation was very effective in destroying Salmonella Typhimurium; however, P. fluorescens was not completely eliminated by electron beam irradiation. Consequently, gamma ray irradiation was more effective than electron beam irradiation in the destruction of P. fluorescens. PMID- 10678419 TI - Microbiological sampling of carcasses by excision or swabbing. AB - Groups of 25 carcasses were obtained by random selection of carcasses at the end of each of eight commercial processes for the dressing or cooling of carcasses. Samples were collected from six groups of pig or beef carcasses by excision or swabbing with sponge, gauze, or cotton wool, with one sample obtained by each of the four methods from a separate, randomly selected site on each carcass. Total aerobic counts, coliforms, and Escherichia coli from each sample were enumerated. Values for the mean log10, log10 mean, and log10 total numbers recovered were calculated for each set of total aerobic counts. Those statistics indicated that the numbers of bacteria recovered by excision or swabbing with sponge or gauze were similar, while the numbers recovered by swabbing with cotton wool were at the lower end of or below the range of the numbers recovered by the other methods. The numbers of coliforms or E. coli recovered from carcasses by sampling areas up to 100 cm2 were too few for the estimation of log mean numbers. Sampling of two groups of carcasses by swabbing with gauze indicated that each 10-fold increase in the area sampled, from 10 to 1,000 cm2, approximately doubled the number of samples from which coliforms or E. coli were recovered. Sampling of six groups of carcasses from one process indicated that the sizes of swabs and volumes of diluent used for processing swabs did not have to be increased proportionally to the area of carcass surface sampled to recover numbers of E. coli proportional to the sampled area. It therefore appears that carcass sampling techniques can be varied widely without compromising the recovery of bacteria, and that the relative efficiencies with which bacteria are recovered by different techniques can be assessed by sampling each carcass in a group of 25 by each of the methods to be compared. PMID- 10678420 TI - Attempts to isolate Helicobacter from cattle and survival of Helicobacter pylori in beef products. AB - This study focused on important factors related to the potential of cattle and beef products to transmit Helicobacter pylori to humans. Mucosal samples were collected from the rumen and abomasum of 105 cattle and were plated on a selective medium to isolate Helicobacter spp.; none of the samples examined contained these bacteria. Studies were also conducted to determine how long H. pylori survives in refrigerated or frozen ground beef; results indicated that the microorganism dies rapidly in ground beef, whether refrigerated or frozen. Packaging in vacuum or air had little effect on survival of the organism. The number of H. pylori decreased in refrigerated samples from 3.3 log10 CFU/g on day 0 to 1.4 log10 CFU/g on day 6. H. pylori died even more rapidly when frozen, decreasing from 3.3 log10 CFU/g on day 0 to 0.5 log10 CFU/g on day 6. Retail beef cuts (n = 20) were also examined for the presence of H. pylori by direct plating on a selective medium and by incubation in an enriched broth followed by plating on a selective medium. None of the retail samples contained H. pylori. This research suggests that transmission of H. pylori from beef and beef products is not a primary factor in the high prevalence of this bacterium in humans. PMID- 10678421 TI - Ribotype analysis of strain distribution in Listeria monocytogenes. AB - Changes in the temporal and spatial patterns of strain distribution for the foodborne pathogen Listeria monocytogenes were studied by ribotyping using the Qualicon Riboprinter system. Ribotype patterns were obtained by using the restriction enzymes EcoRI and PvuII for 72 isolates of L. monocytogenes recovered from smoked salmon samples over a period of 3 years. Each pattern was classified both by comparison to a pattern library and by comparison among the 72 isolate patterns. Eleven EcoRI-based ribogroups and 16 PvuII groups were identified. Eight of the 11 EcoRI ribogroups were found in isolates obtained over a period of >12 months, and 75% of the EcoRI ribogroups that were found in more than one food sample were distributed nationally. Within the set of isolates, there were 26 instances where more than one isolate was obtained from a single food sample. In 35% of these instances, the co-isolates produced different ribotype patterns, indicating that multiple strains of L. monocytogenes commonly coexist in the same environment. Overall, these data indicate that the population of L. monocytogenes consists of a number of widely dispersed strains with little geographic or temporal stratification. PMID- 10678422 TI - Incidence and characterization of Listeria monocytogenes from domestic and imported foods in Korea. AB - A total of 1,537 domestic and imported food products were examined for the incidence of Listeria monocytogenes between 1993 and 1997 in Korea. L. monocytogenes was detected using the U.S. Department of Agriculture isolation method. Isolated L. monocytogenes was confirmed by polymerase chain reaction with hly1 and hly2 primers designed from the listeriolysin O. Overall, 122 samples (7.9%) contained L. monocytogenes. The rate of isolation was 4.3% for beef, 19.1% for pork, 30.2% for chicken, 1.2% for shellfish, 4.4% for raw milk, 4.4% for frozen smoked mussels, and 6.1% for ice cream. No L. monocytogenes was found in pasteurized milk, pasteurized processed cheese, saltwater fish, dried seafoods, or ham. The overall incidence was lower than that reported in previous studies from other countries. Most isolates were serotype 1/2b except for chicken, in which serotype 1/2a was predominant. The serotyping results might imply the presence of food or geography-specific L. monocytogenes strains. PMID- 10678423 TI - Estimating the survival of Clostridium botulinum spores during heat treatments. AB - A recently published study of the inactivation of Clostridium botulinum spores at various temperatures in the range of 101 to 121 degrees C and neutral pH revealed that their semilogarithmic survival curves all had considerable upward concavity. This finding indicated that heat inactivation of the spores under these conditions did not follow a first-order kinetics and that meaningful D values could not be calculated. The individual survival curves could be described by the cumulative form of the Weibull distribution, i.e., by log S = -b(T)t(n(T)), where S is the survival ratio and b(T) and n(T) are temperature-dependent coefficients. The fact that at all temperatures in the above range n(T) was smaller than 1 suggested that as time increases sensitive members of the population parish and survivors with increasing resistance remain. If damage accumulation is not a main factor, and the inactivation is path independent, then survival curves under monotonously increasing temperature can be constructed using a relatively simple model, which can be used to calculate the spores' survival in a limiting case. This is demonstrated with computer-simulated heating curves and the experimental constants of the C. botulinum spores, setting the number of decades reduction to 8, 10, and 12 (the current criterion for commercial sterility). PMID- 10678424 TI - Oyster preservation by high-pressure treatment. AB - The purpose of this study was to analyze the effect of 10-min continuous pressure and pulsed pressure in two 5-min steps (400 MPa at 7 degrees C) on the microbial flora, total volatile bases, pH, and texture of purified and unpurified oysters. High-pressure treatment reduced the number of all the target microorganisms (total viable count, H2S-producing microorganisms, lactic acid bacteria, Brochothrix thermosphacta, and coliforms), in some cases by around 5-log units. The difference between the counts in the control and the pressurized oysters remained stable throughout 41 days of storage at 2 degrees C. No Salmonella spp. were detected in either the control batch or the pressurized batches during this storage period. Deterioration of the oyster was accompanied by increased total volatile bases, mainly in the nonpressurized samples. The pH was practically constant in the pressurized oysters and fell slightly in unpressurized samples. As for mechanical properties, shear strength values were higher in pressurized than in unpressurized oysters. Step-pulse pressurizing (400 MPa at 7 degrees C in two 5-min pulses) produced no apparent advantages over continuous pressurizing based on any of the indices used. PMID- 10678425 TI - The antimicrobial properties of chitosan in mayonnaise and mayonnaise-based shrimp salads. AB - The potential for using chitosan glutamate as a natural food preservative in mayonnaise and mayonnaise-based shrimp salad was investigated. Mayonnaise containing 3 g/liter of chitosan combined with acetic acid (0.16%) or lemon juice (1.2 and 2.6%) was inoculated with log 5 to 6 CFU/g of Salmonella Enteritidis, Zygosaccharomyces bailii, or Lactobacillus fructivorans and stored at 5 and 25 degrees C for 8 days. In mayonnaise containing chitosan and 0.16% acetic acid, 5 log CFU/g of L. fructivorans were inactivated, and numbers remained below the sensitivity limit of the plate counting technique for the duration of the experiment. Z. bailii counts were also reduced by approximately 1 to 2 log CFU/g within the first day of incubation at 25 degrees C, but this was followed by growth on subsequent days, giving an overall growth delay of 2 days. No differences in counts of Z. bailii in mayonnaise stored at 5 degrees C or of Salmonella Enteritidis stored at either temperature were observed. In mayonnaise containing lemon juice at both 1.2 and 2.6%, no substantial differences were observed between the controls and the samples containing chitosan. In shrimp salads stored at 5 degrees C, the presence of a coating of chitosan (9 mg/g of shrimp) inhibited growth of the spoilage flora from approximately log 8 CFU/g in the controls to log 4 CFU/g throughout 4 weeks. However, at 25 degrees C, chitosan was ineffective as a preservative. The results demonstrated that chitosan may be useful as a preservative when combined with acetic acid and chill storage in specific food applications. PMID- 10678426 TI - Microbial testing methods for detection of residual cleaning agents and disinfectants-prevention of ATP bioluminescence measurement errors in the food industry. AB - The ATP luminescence measurement is based on the presence of an enzymatic reaction and may significantly be affected by cleaning agents and disinfectants. In addition, disinfectants can also reduce the activity of the luciferase enzyme and also act as ATP-releasing agents. The agents disrupt the cell walls but preserve ATP in measurable form, and therefore correlation with culture methods can be poor. Therefore, if a rapid method is used to detect ATP, a control must be used for reliable results. The possible effect of disinfectants can be eliminated with a rapid test to minimize sources of error. In the present study a microbiological residue testing method that is nonspecific for residues was developed. The effects of a total of 38 commercial cleaning agents and disinfectants of various types were assessed using two microbiological methods, the Vibrio fischeri photobacteria test and Micrococcus luteus inhibition zone technique. The results show that the V. fischeri photobacteria test is very sensitive. This test can therefore be used for testing cleaning agent residues on surfaces in very small amounts. A small study was also carried out in a food factory to show applicability in processing facilities. The study showed, that a need for this type of method exists in food processing. PMID- 10678427 TI - Characterization of whey cheese packaged under vacuum. AB - Vacuum packaging was assayed at 4 degrees C and was tested in comparison to unpackaged counterparts, in both microbiological and physicochemical terms, in studies pertaining to the preservation of Requeijao, a traditional Portuguese whey cheese. Bacteria were absent (i.e., <10 CFU/g) in whey cheeses on the day of manufacture as a result of thermal processing. After storage, both unpackaged and packaged cheeses exhibited high viable counts of Bacillus, Pseudomonas, Enterobacteriaceae, and lactic acid bacteria (especially lactococci). Yeasts, staphylococci, enterococci, and spore-forming clostridia were severely inhibited by the package vacuum combined with the increasing acidification developed therein. Whey cheeses packaged under vacuum underwent substantial acidification, slight depletion of lactose, and no significant variation in moisture content or texture; conversely, unpackaged whey cheeses exhibited substantial loss of water and a concomitant increase in rigidity. Vacuum packaging strongly inhibited lipolysis (even if viable counts of some microbial groups were high); saturated fatty acids (mainly C16:0 and C14:0) accounted for ca. 73% of the total free fatty acid content, whereas the most concentrated unsaturated fatty acids were C18:1 and C18:2 (ca. 14% each). The conclusions generated in our study are, in general, useful for a wide range of whey cheeses worldwide: i.e., Requeson (Spain), Ricotta (Italy), Broccio (France), and Anthotyro (Greece). In addition, our conclusions are particularly helpful in terms of improving the safety of Requeijao, a widely acclaimed dairy specialty. PMID- 10678428 TI - The boundary for growth of Zygosaccharomyces bailii in acidified products described by models for time to growth and probability of growth. AB - Models to predict days to growth and probability of growth of Zygosaccharomyces bailii in high-acid foods were developed, and the equations are presented here. The models were constructed from measurements of growth of Z. bailii using automated turbidimetry over a 29-day period at various pH, NaCl, fructose, and acetic acid levels. Statistical analyses were carried out using Statistical Analysis Systems LIFEREG procedures, and the data were fitted to log-logistic models. Model 1 predicts days to growth based on two factors, combined molar concentration of salt plus sugar and undissociated acetic acid. This model allows a growth/no-growth boundary to be visualized. The boundary is comparable with that established by G. Tuynenburg Muys (Process Biochem. 6:25-28, 1971), which still forms the basis of industry assumptions about the stability of acidic foods. Model 2 predicts days to growth based on the four independent factors of salt, sugar, acetic acid, and pH levels and is, therefore, much more useful for product development. Validation data derived from challenge studies in retail products from the U.S. market are presented for Model 2, showing that the model gives reliable, fail-safe predictions and is suitable for use in predicting growth responses of Z. bailii in high-acid foods. Model 3 predicts probability of growth of Z. bailii in 29 days. This model is most useful for spoilage risk assessment. All three models showed good agreement between predictions and observed values for the underlying data. PMID- 10678429 TI - Effects of substrate, water activity, and temperature on growth and verrucosidin production by Penicillium polonicum isolated from dry-cured ham. AB - Penicillium polonicum, a common mold on dry-cured meat products, is able to produce verrucosidin, a potent neurotoxin. The ability of P. polonicum isolated from dry-cured ham to grow and produce verrucosidin from 4 to 40 degrees C at water activities (a(w)) of 0.99, 0.97, and 0.95 on malt extract agar (MEA) and a medium made up with meat extract, peptone, and agar (MPA) was evaluated. Verrucosidin was quantified by high-pressure liquid chromatography and mass spectrometry. P. polonicum was able to grow on MEA and MPA at all the a(w) values tested from 4 to 37 degrees C but not at 40 degrees C. The optimal environmental conditions for growth were 20 degrees C, 0.99 a(w) on MEA and 20 to 25 degrees C, 0.97 a(w) on MPA, but the highest amount of verrucosidin was obtained at 25 degrees C, 0.99 a(w) in both media. No direct correlation between extension of mold growth and verrucosidin production was found. Temperature appears to be the most important factor ruling mycelial growth, whereas verrucosidin accumulation is mostly influenced by a(w). However, analysis of variance of the data showed that there was a complex interaction among all the environmental factors (medium, temperature, and a(w)) that significantly (P < 0.0001) affected growth and verrucosidin production. The reduction of a(w) to intermediates values of 0.95 has a stronger effect on growth on MEA than on MPA. Given that the meat-based medium proved to be an appropriate substrate for the biosynthesis of verrucosidin by P. polonicum, the ability of this mold to produce the toxin on meat products should be established. PMID- 10678430 TI - Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages. AB - The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied. The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch). The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage. Tyramine, cadaverine, and putrescine were the main amines formed during the ripening. The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated. A great reduction in tyramine content was achieved when L. sakei CTC494 was inoculated, whereas L. curvatus CTC371 only attenuated tyramine accumulation compared with the control batch. Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora. Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C. On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage. The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines. PMID- 10678431 TI - Histamine and biogenic amine production by Morganella morganii isolated from temperature-abused albacore. AB - Histamine-producing bacteria were isolated from albacore stored at 0, 25, 30, and 37 degrees C. They were screened using Niven's differential medium, and their histamine production was confirmed by high-pressure liquid chromatography analysis. The optimum temperature for growth of histamine-producing bacteria was 25 degrees C. The bacterium producing the highest level of histamine was isolated from fish abused at 25 degrees C. It was identified as Morganella morganii by morphological, cultural, biochemical, and antimicrobial characteristics and by the Vitek microbial identification system. The M. morganii isolate was inoculated into tuna fish infusion broth medium, and the effect of temperature was determined for microbial growth and formation of histamine and other biogenic amines. The isolate produced the highest level of histamine, 5,253 ppm, at 25 degrees C in the stationary phase. At 15 degrees C, histamine production was reduced to 2,769 ppm. Neither microbial growth nor histamine formation was detected at 4 degrees C. To determine whether the isolate can also produce other biogenic amines that can potentiate histamine toxicity, production of cadaverine, putrescine, serotonin, tryptamine, tyramine, phenylethylamine, spermidine, and spermine by the isolate was also monitored. Cadaverine, putrescine, and phenylethylamine were detected with microbial growth in the tuna fish infusion broth medium. The optimum temperature for cadaverine, putrescine, and phenylethylamine formation was found to be 25 degrees C, as it was for histamine. PMID- 10678432 TI - A sandwich enzyme-linked immunosorbent assay for the detection of almonds in foods. AB - An enzyme-linked immunosorbent assay was developed to detect almonds as potential allergenic contaminants in food. Polyclonal antibodies directed against roasted almonds were partially purified from immunized sheep and rabbits and used as capture and secondary antibodies, respectively, in a sandwich-type, 96-well plate format. Food samples and almond-spiked samples were extracted 1:10 in phosphate buffered saline at 60 degrees C for 2 h, centrifuged, and applied to wells coated with sheep anti-almond antibody. After incubation, washing, and the addition of rabbit anti-almond antibody, the amount of almond present was detected with the subsequent addition of goat anti-rabbit immunoglobulin G-alkaline phosphatase conjugate and p-nitrophenyl phosphate substrate. Plate absorbances were read at 410 nm, and standard curves were developed in all matrices to quantify unknowns. Antibodies developed were specific for almond; however, some cross-reactivity was observed with extracts of some tree nuts and sesame seeds. Sodium dodecylsulfate polyacrylamide gel electrophoresis and Western immunoblotting indicated that sheep anti-almond antibody recognized proteins extracted from black walnuts, Brazil nuts, cashews, hazelnuts, macadamia nuts, pistachios, and sesame seeds in addition to those from almond. The assay was optimized to detect less than 1 ppm of almond and was used successfully to determine almond residues in cereal and chocolate without cross-reacting interferences. A retail survey of 20 brands of cereal demonstrated that the assay produced statistically consistent results. This assay provides a useful quality control tool for the food industry for the protection of consumers allergic to almonds. PMID- 10678433 TI - Identification of central nervous system tissue in retail meat products. AB - A procedure to detect tissues from the central nervous system that involved quantification of cholesterol and immunochemical detection of neuron-specific enolase and glial fibrillary acidic protein was used to analyze 402 samples of heat-treated meat products from various food outlets in Germany. The cholesterol content of 16 samples (4.0%) indicated the possible presence of central nervous system tissue because the levels exceeded the normal maximum cholesterol content of cooked sausages. In 7 of these 16 heat-treated meat products, immunoblotting of both neuron-specific enolase and glial fibrillary acidic protein confirmed the presence of CNS tissue. Repeated sampling by veterinary officials and analysis by both cholesterol quantification and immunoblotting confirmed these findings. Whereas all of the control samples (with and without added central nervous system tissue) were correctly classified by both cholesterol quantification and immunoblotting, negative results of immunoblotting must be carefully interpreted in the case of intensively heat-treated meat products. Thus, studies have yet to establish an increase in sensitivity of immunoblotting of neuron-specific enolase and glial fibrillary acidic protein. However, the detection of illegal use of central nervous system tissue in heat-treated retail meat products demonstrates the need for suitable analytical methods to control transmissible encephalopathies and to enforce labeling laws. PMID- 10678434 TI - Specificity of a conductance assay for enumeration of Escherichia coli from broiler carcass rinse samples containing genetically similar species. AB - Experiments were conducted to evaluate the specificity of a rapid method for enumeration of Escherichia coli from fresh broiler chicken carcasses. In three separate trials, E. coli, Citrobacter freundii, Salmonella Enteritidis, and Shigella sonnei were serially diluted and then inoculated into identical broiler chicken carcass rinses. Inoculated rinses were mixed with double-strength Coliform Medium supplemented with 2% dextrose. This mixture was placed in a Bactometer module in duplicate, and conductance was measured at 44 degrees C. Results indicated that C. freundii did not grow to an appreciable degree in the selective medium at 44 degrees C. Salmonella Enteritidis grew similarly to E. coli; however, an initial level of 10(6) Salmonella in the food product would be required for Salmonella to interfere with enumeration of E. coli using this method. S. sonnei grew at a more rapid rate than E. coli; however, there was an interaction between the regression lines formed when serial dilutions (log10 CFU/ml) were compared to E. coli detection times for these two species of bacteria. Therefore, high levels of S. sonnei in a food sample may interfere with the enumeration of E. coli. In general, Salmonella and Shigella are not found at high enough levels on poultry products to interfere with enumeration of E. coli using this method and, if found at high levels, would be detected and rejected using this procedure. Hence, the presence of organisms that are genetically and phylogenetically similar to E. coli would not preclude enumeration of E. coli using conductance under these conditions. PMID- 10678435 TI - Validation and analysis of modeled predictions of growth of Bacillus cereus spores in boiled rice. AB - The growth of psychrotrophic Bacillus cereus 404 from spores in boiled rice was examined experimentally at 15, 20, and 30 degrees C. Using the Gompertz function, observed growth was modeled, and these kinetic values were compared with kinetic values for the growth of mesophilic vegetative cells as predicted by the U.S. Department of Agriculture's Pathogen Modeling Program, version 5.1. An analysis of variance indicated no statistically significant difference between observed and predicted values. A graphical comparison of kinetic values demonstrated that modeled predictions were "fail safe" for generation time and exponential growth rate at all temperatures. The model also was fail safe for lag-phase duration at 20 and 30 degrees C but not at 15 degrees C. Bias factors of 0.55, 0.82, and 1.82 for generation time, lag-phase duration, and exponential growth rate, respectively, indicated that the model generally was fail safe and hence provided a margin of safety in its growth predictions. Accuracy factors of 1.82, 1.60, and 1.82 for generation time, lag-phase duration, and exponential growth rate, respectively, quantitatively demonstrated the degree of difference between predicted and observed values. Although the Pathogen Modeling Program produced reasonably accurate predictions of the growth of psychrotrophic B. cereus from spores in boiled rice, the margin of safety provided by the model may be more conservative than desired for some applications. It is recommended that if microbial growth modeling is to be applied to any food safety or processing situation, it is best to validate the model before use. Once experimental data are gathered, graphical and quantitative methods of analysis can be useful tools for evaluating specific trends in model prediction and identifying important deviations between predicted and observed data. PMID- 10678436 TI - Heavy metals in mussels (Mytilus galloprovincialis) from the lonian Sea, Italy. AB - Concentrations of six heavy metals (Hg, Pb, Cd, Cr, Zn, and Sn) were determined in mussels (Mytilus galloprovincialis) collected between June and September 1997 from 10 locations along a sound formed by two inlets (Mar Piccolo) near the Gulf of Taranto (Ionian Sea, Italy). The average concentrations of the heavy metals found in mussels samples were 0.15 mg/kg for Hg, 1.19 mg/kg for Pb, 0.64 mg/kg for Cd, 0.31 mg/kg for Cr, 5.15 mg/kg for Zn, and 0.54 mg/kg for Sn. The concentrations of heavy metals in mussels from the first inlet did not differ greatly from those observed in mussels from the second inlet. The concentrations of heavy metals in the mussels analyzed were below acceptable levels for human consumption. PMID- 10678437 TI - Application of the Bigelow (z-value) model and histamine detection to determine the time and temperature required to eliminate Morganella morganii from seafood. AB - In New Zealand, the product most frequently implicated in cases of scombroid or histamine poisoning is the hot-smoked fish, kahawai (Arripis trutta). A properly controlled heating step in the production of hot-smoked seafood could eliminate bacteria able to convert the amino acid histidine to histamine. In this study, we determined the core temperatures and times required during hot smoking of kahawai to eliminate histamine-forming bacteria and to ensure a final product that will not produce histamine if subsequent temperature abuse occurs. Morganella morganii strains previously isolated from portions of hot-smoked kahawai with elevated histamine levels were inoculated onto product to be tested. A variation of the Bigelow or z-value model was used to generate a thermal death time graph, where the production of histamine, in a heat-treated and subsequently temperature abused sample, was scored as a positive value (growth) and the absence of histamine was scored as a negative value (no growth). From a line fitted to the data, calculated times for the elimination of histamine-forming bacteria at test temperatures of 58, 59, 60, 61, and 62 degrees C were estimated to be 15.27, 8.81, 4.79, 2.68, and 1.46 min, respectively, giving a z value of 3.85 degrees C. This approach to thermal death determination, based on the presence or absence of a bacterial metabolite, proved to be an efficient way to determine the thermal regime required to eliminate bacteria capable of converting histidine to histamine on kahawai. PMID- 10678438 TI - Immunochemical detection of molds: a review. AB - Molds are widely distributed in nature and cause deterioration of foods and feeds. Their mycotoxins can adversely affect human and animal health. Suitable assays for molds, therefore, are required to implement control and regulatory strategies and to develop appropriate feeding regimens for mold-infested feeds. Many different types of mold assays have been used, most of which are not reproducible or accurate. However, the immunoassays, particularly enzyme-linked immunosorbent assays (ELISAs), can be especially useful. Among these, assays that detect the water-soluble extracellular secretions of fungi, the exoantigens, are generally able to detect fungi at the genus or species level, whereas the heat stable polysaccharides tend to be specific for one or more genus of fungi. Several species and genus (genera)-specific ELISAs have been developed using monoclonal or polyclonal antibodies against exoantigens and heat-stable polysaccharides from a wide range of fungi, including Aspergillus, Penicillium, and Fusarium species. Other assays have been developed that nonspecifically detect mold in food or feed, some using antibodies against a mixture of antigens from different fungi. These assays are highly sensitive, are easy to perform, and provide an index of the amount of mold present in the sample. Further refinement of these assays should facilitate their widespread use by food and feed processors, regulatory agencies, taxonomists, and research scientists. PMID- 10678439 TI - Factors influencing clinical outcomes after revascularization in the asymptomatic cardiac ischemia pilot (ACIP). ACIP Study Group. AB - BACKGROUND AND AIM: The Asymptomatic Cardiac Ischemia Pilot is the first randomized trial where revascularization involved choice of either coronary bypass or angioplasty used in an early or a delayed symptom-driven approach. One year outcomes were favorable (reduced recurrent ischemia and adverse outcomes) for an early revascularization strategy (within 4 weeks), compared with an early medical strategy when revascularization was delayed until symptom-driven. This ancillary study examined variables influencing outcomes after these 2 revascularization approaches (early vs. delayed until symptom-driven). METHODS: Participants were clinically stable coronary disease patients with stress-induced and daily life ischemia who underwent revascularization. Characteristics associated with clinical outcomes occurring within the year following revascularization were examined using Cox regression analysis. RESULTS: A total of 262 patients received revascularization; 170 in the early approach and 92 in the delayed symptom-driven approach. Thirty-three patients had adverse outcomes (death, nonfatal myocardial infarction, or repeat revascularization) during 1 year follow-up. The most important independent predictor of improved outcome during the follow-up year was attempted revascularization of > or = 66% of vessels with significant stenosis for the early (risk ratio [RR] 0.25, 95% confidence interval [CI] 0.09-0.67) and the delayed (RR 0.21, CI 0.08-0.58) approaches. Factors such as age, stress test results, and coronary angiographic findings did not predict clinical outcome. CONCLUSIONS: Our findings are important in the planning of a large trial with longer follow-up. PMID- 10678440 TI - Self-expanding endovascular stent-graft implant for treatment of descending aortic diseases. AB - BACKGROUND: Aneurysms and dissections involving the descending thoracic aorta and the distal portion of the aortic arch are difficult to resolve surgically. The introduction of endovascular self-expanding stent-grafts has simplified the operation. Given the complications associated with their peripheral placement, we explored the feasibility of surgical insertion. METHODS: Thirteen patients underwent surgical insertion of a stent-graft into the aortic arch via longitudinal aortotomy. Six patients had aneurysms (ruptured in two, and seven dissections (acute in two, ruptured in one). Five patients also underwent associated procedures including aortic valve replacement (one), ascending aorta replacement (two), arch replacement (one), and coronary artery bypass (one). RESULTS: There was one intraoperative death due to ascending aortic dissection, and two hospital deaths due to multiple complications. Of ten patients discharged, one died 3 months postoperatively. The remaining survivors are well, and imaging studies confirmed adequate correction of the aortic disease. CONCLUSIONS: The use of this technique simplifies the operation and treatment of particular cases of aortic disease. The observed morbidity and mortality are due to factors independent of the technique. PMID- 10678441 TI - Clinical results of endarterectomy of the right and left anterior descending coronary arteries. AB - In this study, we examined the clinical outcome of coronary endarterectomy. From 1990 to 1998, 4839 patients underwent surgical revascularization. Coronary artery bypass graft surgery (CABG) was performed alone on 4516 patients, was combined with right coronary artery endarterectomy (RCA-E) in 242 patients, and was combined with left anterior descending coronary artery endarterectomy (LAD-E) in 81 patients. An analysis of preoperative variables revealed a higher proportion of males (90.7% vs 80.2%, p < 0.001), of patients with low ejection fraction (< 35%; 4.6% vs 1.7%, p < 0.001), and of three-vessel disease (47.9% vs 36%, p < 0.001) in the RCA-E versus the CABG patients. There was a higher proportion of unstable angina (51.9% vs 40.3%, p = 0.04) in the LAD-E patients. The 30-day mortality rate for CABG was 2% versus 2.5% for RCA-E and 3.7% for LAD-E (p = NS). Perioperative myocardial infarction (MI) rate for CABG was 3.4% versus 7.0% for RCA-E (p < 0.001) and 4.9% for LAD-E patients (p = NS). Postoperative low cardiac output syndrome was recorded in 11.5% of CABG, 18.6% of RCA-E (p = 0.01), and 11.1% of LAD-E (p = NS) patients. Predictors of postoperative bad outcome (death, MI, low cardiac output, cerebrovascular accident) were preoperative intra-aortic balloon pump, repeat operation, ejection fraction of < 35%, renal insufficiency, female gender, RCA-E, and age over 70. Protective factors included the use of internal mammary artery, multiple arterial grafts, and warm cardioplegia. Actuarial analysis at 6, 12, and 24 months showed late mortality rates of 0.8%, 1.3%, and 2.1% for CABG; 1.2%, 3.7%, and 3.7% for RCA-E; and 2.9%, 2.9%, and 2.9% for LAD-E, respectively. Late MI occurrence was 0.4%, 0.4%, and 0.7% for CABG; 1.5%, 1.5%, and 2.7% for RCA-E; and 0% for LAD-E, respectively. Multivariate analysis found renal insufficiency, ejection fraction of < 35%, repeat operation, female gender, New York Heart Association functional class IV, and diabetes to be predictors for late adverse events (recurrence of angina, MI, and cardiac death), and RCA-E was found to be a predictor of late MI. We conclude that the use of coronary endarterectomy to achieve complete revascularization in patients with diffuse distal coronary artery disease is a reasonable option, associated with a minimal addition in complication rates. PMID- 10678442 TI - Use of extrafascially harvested radial artery for coronary artery revascularization: technical considerations. AB - BACKGROUND: The use of the radial artery for coronary artery revascularization was abandoned due to its tendency for spasm; the revival was attributed to improved harvesting technique as well as the use of calcium channel blockers. METHODS: Between February 1996 and June 1997, the radial artery graft was used in 77 of 89 consecutive patients undergoing coronary artery bypass graft surgery. Only the patients with positive Allen's test or forearm deformity were denied the use of the radial artery. We used an extrafascial, no-touch technique using low strength electrocautery for harvesting the radial artery. Calcium channel blockers were not used in any of these patients. RESULTS: There were no early deaths. No patient sustained perioperative myocardial infarction or required intra-aortic balloon pump. Only one patient required inotropic agents. Three noncardiac late deaths occurred during the follow-up of 6 to 24 months. No early or late ischemic or functional forearm disability was reported in any of the patients. CONCLUSIONS: The radial artery is easy to harvest and safe to use routinely. When harvested extrafascially, diltiazem infusion may not be necessary. Maximal arterial-global revascularization using the left internal thoracic artery-to-left anterior descending coronary artery and radial artery-to circumflex artery system may improve the early and long-term results. PMID- 10678443 TI - Excision of coronary artery fistula and coronary artery reconstruction without cardiopulmonary bypass. AB - In three patients, coronary artery fistulas originating from a conal branch of the mid-segment of the left anterior descending coronary artery (n = 2) and right coronary artery (n = 1) with drainage into the right atrium (n = 2) and right ventricle (n = 1) were successfully closed without the use of cardiopulmonary bypass. The use of a coronary artery stabilizer greatly facilitated the operation by immobilization of the fistula, its supplying coronary artery, and the regional myocardium. In selected patients, this technique allows secure closure of the fistula and meticulous reconstruction of the coronary artery without the use of cardiopulmonary bypass. PMID- 10678444 TI - New horizons on the surgical treatment of dilated cardiomyopathy. PMID- 10678445 TI - Left ventricular volume reduction: new dawn or false horizon? Basic science and clinical doubts. AB - Left ventricular volume reduction surgery (LVVR) for end-stage dilated cardiomyopathy is a surgical option used selectively but with unclear long-term results. Increasing numbers of reports are appearing in the literature. These should be pooled into an international registry through collaborative efforts that allow for more effective analysis. Furthermore, high priority must be given to identify subgroups of patients who will potentially gain most benefit from LVVR. Basic science may add invaluable data and in this article we describe how intraoperative myocardial biopsies from patients with idiopathic dilated cardiomyopathy were utilized to isolate myocytes in an effort to determine differential physiological characteristics at the cellular level. The result showed various degrees of contractile anomalies in response to electrical stimulation associated with defective calcium handling as reflected by measurements of calcium transients. It is hoped that this approach may be extended to preoperative catheter biopsy to gain information that will facilitate patient selection to avoid unnecessary surgical failures. PMID- 10678446 TI - Endoventricular patch reconstruction in large ischemic wall-motion abnormalities. AB - Endoventricular patch plasty (EVPP) has been used since 1984 to rebuild the left ventricle. The global experience of our group includes more than 835 cases. Large wall-motion abnormalities were detected by the center line method when > 60% of the circumference of the left ventricle was asynergic. In this series, 269 patients had an ejection fraction < 30%. Surgery for repair of large wall-motion abnormalities was conducted on the arrested heart with insertion within the left ventricle of a patch rebuilding the contractile area while leaving a residual volume between 50 and 70 cc/m2 of body surface. The global results of the technique of EVPP are analyzed on the last 700 operated patients. Three series of patients with large wall-motion abnormalities were examined. We conclude that this technique is appropriate in advanced stages of ischemic disease as an alternative to cardiac transplant. At an operative risk of approximately 12%, improvement is obtained in 80% of cases. PMID- 10678447 TI - Commonality of ischemic and dilated cardiomyopathy: laplace and ventricular restoration. PMID- 10678448 TI - Left ventricular volume reduction for end-stage heart disease. AB - Partial left ventriculectomy (PLV) was recently introduced for end-stage dilated cardiomyopathy to improve ventricular function. Since November 1996 we have performed PLV in 14 patients; preoperatively 4 patients had idiopathic dilated cardiomyopathy and 10 had ischemic dilated cardiomyopathy. 57.1% of patients were in New York Heart Association functional Class IV. The mitral valve was replaced in 11 patients. Postoperative echocardiography showed a reduction of left end diastolic diameter (55.4 +/- 5.4 mm) and an increase in forward ejection (cardiac index from 2.19 +/- 0.571 min/m2 to 2.67 +/- 0.931/min/m2). The 30-day mortality was 28.6% and 20-month survival was 57.2%. Only one patient was not in NYHA functional class due to postoperative progressive mitral incompetence. Prognostic factors should be identified to avoid early failure. However, even if the mortality rate for PLV high, this operation is a valid choice for the treatment of end-stage dilated cardiomyopathy. PMID- 10678449 TI - Dilated cardiomyopathy: changing pathophysiological concepts and mechanisms of dysfunction. AB - Experimental observations made over the past two decades have led to a profound shift in the conceptual paradigms about the syndrome of heart failure and dilated cardiomyopathy. As a consequence, heart failure is currently considered a complex disease and is not merely characterized by hemodynamic disturbances. It is now believed that the syndrome is governed and impelled by neurohormonal imbalances and intracardiac paracrine processes. The latter processes are mediated by activated cardiac endothelial cells and cytokines, creating a state of cardiac maladaption and leading to disease progression. Therapeutic interventions such as operative left ventricular volume reduction or mitral valve reconstruction should therefore no longer be solely interpreted in terms of hemodynamics (i.e., symptomatic improvements). Effects on neurohormonal, endothelial, and cytokine activities should be taken equally into account. PMID- 10678450 TI - Low vision and blindness. PMID- 10678451 TI - Outcome assessment of the rehabilitation of the visually impaired. AB - The purposes of this project are to establish the psychometric properties of instruments used to gather data relevant to blind rehabilitation outcomes, to refine the scaling and scoring protocols for the instruments, and to revise and refine the outcome instruments. This 3-year project will gather outcome and demographic data from an estimated 1,200 visually impaired veterans per year, along with a companion sample an estimated 1,200 visually impaired nonveterans per year, using the following core measures: Blind Rehabilitation Service Follow up Outcome Survey (BRSFOutSur) measuring functional performance, Blind Rehabilitation Service Data Base (BRSDBase) recording subject characteristics, and Blind Rehabilitation Service Satisfaction Survey (BRSSatSur) measuring satisfaction with rehabilitation. As of July 1999, data from 2,624 veterans have been collected for the demographic instrument, from 1,630 veterans for the functional outcomes instrument, and from 1,655 veterans for the satisfaction instrument. Data collection and analysis are currently ongoing. These findings and the further development of outcome instruments in this area will contribute to greater efficiency and effectiveness of the delivery of blind rehabilitation services by the Department of Veterans Affairs. PMID- 10678452 TI - Characteristics of AMD patients with low vision receiving visual rehabilitation. AB - The purpose of this retrospective study done on 255 AMD patients evaluated at a low vision rehabilitation service was: 1) to describe the visual function characteristics (VFCs) of AMD patients presenting to visual rehabilitation, 2) to document changes in these VFCs between initial and follow-up rehabilitation visits, and 3) to investigate the relationship of the VFCs found at rehabilitation intake to the length of time between initial diagnosis and initial rehabilitation visit. Standard clinical testing (visual acuity and contrast sensitivity) as well as Scanning Laser Ophthalmoscope (SLO) visual function testing were performed to determine visual function including: 1) macular perimetry for scotoma boundary mapping and 2) PRL (preferred retinal locus) location and abilities in fixation, saccadic, and pursuit eye movements. The difference between the first and second visit VFCs were compared to the length of time between visits for 44 of the 255 patients returning for a second visit 0.5 to 4.5 years later. Finally, the initial date of AMD diagnosis was found for 51 of the 255 patients to analyze VFCs as a function of the time duration between diagnosis and the intake to the rehabilitation. Most VFCs had a wide range of results at initial intake to rehabilitation, while all patients had significant visual impairment by 24 months after initial diagnosis. The majority of low vision patients with AMD have bilateral central scotomas with the corresponding visual function and ADL problems that can often be overcome with visual rehabilitation. PMID- 10678453 TI - Visual factors and mobility in persons with age-related macular degeneration. AB - The objectives of this study were to determine the effects of reducing light level on mobility performance in persons with age-related macular degeneration (ARMD) and how performance relates to measures of visual sensory and perceptual function. ARMD results in the loss of central, high-acuity vision and is the leading cause of vision loss in veterans participating in the blind rehabilitation programs of the Department of Veterans Affairs. In 41 subjects with ARMD acuity, peak letter contrast sensitivity, visual field extent, glare disability, color confusion, spatio-temporal contrast sensitivity, motion sensitivity, scanning ability, and figure-ground discrimination were measured to determine their ability to predict mobility performance. Mobility performance was assessed under photopic (high illumination) and mesopic (low illumination) lighting conditions on a laboratory obstacle course and two real-world courses, an indoor hallway and an outdoor residential route. Reducing illumination resulted in significant increases in the time to complete each course and the number of mobility incidents (errors) that occurred. Two measures of overall performance, total time and total mobility incidents, were calculated for each course by summing time and incidents over the two illumination levels. Combinations of vision variables were able to account for 30 to 60% of the variance in the measures of overall performance. Log contrast sensitivity measured with the Pelli-Robson chart test and visual field extent were the most important predictors of performance. Other variables making significant contributions to prediction in multi-predictor models included: scanning ability, glare sensitivity, color confusion, and peak contrast sensitivity to drifting gratings. PMID- 10678454 TI - Auditory perception of walls via spectral variations in the ambient sound field. AB - Individuals with visual disabilities often use their hearing in order to maintain a line of travel parallel to walls, such as when walking down a hallway or along the side of a building. Previous studies established that this ability depends on the sense of hearing, but the specific acoustic information has not been investigated. The present paper describes a model of how sound pressure builds up within a meter or so in front of a wall, particularly in the low frequency end of the sound spectrum. This buildup of sound pressure is based on ambient or "background" sound, not self-produced sound such as footsteps. The model leads to a prediction that walls are detected by means of a spectral shift toward low frequencies. This prediction was tested in three experiments, in which sighted adults listened for such spectral shifts. In each experiment, a threshold value was obtained corresponding to the farthest simulated distance from a wall that could be detected. Threshold values were in good agreement with previous observations of the distance at which pedestrians can utilize acoustic information from walls. There was no evidence that simulated listener motion enhanced perception of walls. The model underlying these experiments implies that the term echolocation carries inappropriate connotations about the auditory processes that are involved in walking along walls. It is suggested that a more apt description is that pedestrians listen for spatial variations in the structure of the ambient sound field. PMID- 10678455 TI - A biomechanical evaluation of visually impaired persons' gait and long-cane mechanics. AB - This study was designed to compare selected kinematic components of gait and long cane mechanics between groups of visually impaired travelers. Twenty subjects were placed in Traditional or Modified technique groups according to their long cane traveling technique. Subjects were measured during the following conditions; 1) normal walking (NW), 2) walking while anticipating a simulated drop-off (AD), 3) walking while responding to an audible task (ST) and, 4) walking while anticipating a simulated drop-off and responding to an audible task (STAD). Data were analyzed using a repeated measures analysis of variance (ANOVA) and Pearson's r correlation coefficient. Analyses revealed no differences between groups of travelers. However, significant differences were noted between trials for components of gait velocity, stride length, and hip flexion velocity. These findings may indicate a potentially dangerous alteration in the normal gait cycle of visually impaired travelers when faced with additional attention-demanding tasks while walking. PMID- 10678456 TI - The use of the sonic pathfinder as a secondary mobility aid for travel in business environments: a single-subject design. AB - Two studies were conducted using a single subject research design in different business environments to evaluate the efficacy of the Sonic Pathfinder for increasing efficiency in travel with an experienced and accomplished blind traveller. The Sonic Pathfinder is one of eight or nine commercially available electronic travel aids (ETAs) designed for use by blind or visually impaired persons. The use of ETAs are thought to result in more rapid travel and a greater ability to detect and avoid obstacles in one's path. Elapsed time for travel and the number of unintentional contacts made while travelling were used as the dependent variables in both studies. No marked effect was observed for either variable in either study. An additional traveller who was slower and more tenuous in his movements was recruited to participate in the second study to investigate the possibility that a floor effect may have masked the results of the use of the ETA with the first subject. However, no marked effects were observed with this subject either. Yet, both subjects stated that the use of the aid increased their distance for environmental preview over and above that provided by the long cane alone, while providing them with the opportunity to judge the distance of objects approached beyond cane length. Discussion centers on the need to identify other means to evaluate the utility of this aid in light of the positive statements made by the participants of these studies. PMID- 10678457 TI - Remote infrared signage evaluation for transit stations and intersections. AB - Opportunities for education and employment depend upon effective and independent travel. For mainstream society, this is accomplished to a large extent by printed signs. People who are print disabled, visually impaired, or totally blind are at a disadvantage because they do not have access to signage. Remote infrared signage, such as the Talking Signs (TS) system, provides a solution to this need by labeling the environment for distant viewing. The system uses a transmitting "sign" and a hand-held receiver to tell people about their surroundings. In a seamless infrared signage environment, a visually impaired traveler could: walk safely across an intersection to an ATM or fare machine, from fare machine to bus stop, from bus stop to bus; from bus to building, from building to elevator, from elevator to office, from office to restroom, and so forth. This paper focuses on two problems that are among the most challenging and dangerous faced by blind travelers: negotiating complex transit stations and controlled intersections. We report on human factors studies of TS in these critical tasks, examining such issues as how much training is needed to use the system, its impact on performance and safety, benefits for different population subgroups and user opinions of its value. Results indicate that blind people can quickly and easily learn to use remote infrared signage effectively, and that its use improves travel safety, efficiency, and independence. PMID- 10678458 TI - Relative locations of macular scotomas near the PRL: effect on low vision reading. AB - Patients referred for low vision rehabilitation had Minnesota Reading Acuity (MNRead), visual acuity (VA), and scanning laser ophthalmoscope (SLO) macular function testing performed in their initial evaluation to determine whether dense macular scotomas near the preferred retinal locus (PRL) have a significant effect on the characteristics of reading based on rate. The 99 subjects had macular scotoma characteristics relative to the fovea/PRL of: 22% only to the right; 15% only to the left; 26% both the right and left; 19% above or below; 17% had no dense scotomas. Reading performance (maximum reading speed, critical print size, and reading acuity) was significantly different between the non-scotoma group and all of the scotoma groups. There was no statistically significant difference in the characteristics of reading based on rate between the four scotoma groups: within each there was a wide variation in the characteristics of reading based on rate not fully explained by either VA or scotoma location. The position of the scotoma relative to the PRL was not a statistically significant factor in determining reading rate as found in studies on normally sighted people with artificial scotomas. Other factors (e.g., maybe PRL ability in fixation and saccadic eye movements and/or cognitive ability) are significantly involved in determining reading rate characteristics in people with macular scotomas. PMID- 10678459 TI - Impact of digital miniaturization and networked topologies on access to next generation telecommunication by people with visual disabilities. AB - In the past, telecommunication technologies did not present any particular problem for persons with visual disabilities. The telephones themselves were auditory in nature and could be operated by touch. As telecommunication begins to incorporate video and as telecommunication devices become more complex (including the incorporation of visual displays), new barriers are appearing. Fortunately, advancing technologies are also providing new opportunities for access. The rapidly shrinking size and cost of electronics is allowing us to build intelligence and flexibility into telecommunication products. Advances will soon allow voice to be incorporated into most devices. In addition, clever use of networks and network-based services will allow access features to be built directly into the network, providing access to key visual information. As a result, future telecommunication systems can be more accessible than technologies of the past-if they are implemented correctly. PMID- 10678460 TI - Subretinal implantation of semiconductor-based photodiodes: progress and challenges. AB - Retinal diseases that result in photoreceptor degeneration may spare the inner retinal layers. This review concerns a prosthetic approach to restoring visual function through the use of a semiconductor-based microphotodiode array implant, designed to be placed under the neural retina in the subretinal space. The fundamental idea is that current generated by the device in response to light stimulation will alter the membrane potential of overlying neurons and thereby activate the visual system. Initial acute studies indicated that the implant will function in the subretinal space in the absence of an external power supply. More recent and ongoing studies involve chronic subretinal implantations in normal animals. Post-operative studies have demonstrated that implant function will persist for many months. These chronic studies have also assessed the biocompatibility of the implant. Photoreceptors are lost directly overlying the implant, due to the blockade of choroidal circulation to the outer retina by the solid disk device. In comparison, the inner retina maintains its characteristic lamellar structure. Away from the implant site, the retina retains normal anatomy and function. Future studies are needed to determine whether the implant can establish a functional connection to the inner retina and to determine the quality of this connection. PMID- 10678461 TI - A new method for continuous, long-term polysomnographic recording of neonatal rats. AB - STUDY OBJECTIVES: Many findings suggest that in altricial mammals neonatal REM sleep has developmental functions. However, investigations of these developmental functions has been hampered by technical limitations of the conventional polysomnographic (PSG) recording technique. One limitation is that continuous (24 hour/day), long-term (weeks) PSG recordings have not been achieved. A second limitation is that the metal screw electrodes and head plugs cemented to the skull cannot be removed to allow the neonate to mature into adulthood. As a result of these limitations, the relationship between neonatal sleep/wake variables and adult variables has not been studied. Also the effects of polysomnographically controlled neonatal REM sleep deprivation on adult variables have not been studied. The present work describes a new technique called the soft head plug (SH) method for continuous, long-term PSG recording. DESIGN: In the new technique, electrodes are thin, strong, Teflon wires that are led by a suturing needle through the soft skull to the epidural space, then with a U-turn exited from the skull and tied to the entry wire. Thus, in contrast to the conventional technique, the soft head plug technique does not use screws as electrodes and does not cement a hard, relatively large electrode plug to the skull, removal of which is fatal or very traumatic. The SH recording electrodes can be removed without damage to neonates. SETTING: NA. PATIENTS: NA. INTERVENTIONS: NA. RESULTS: In the present study sleep/wake results with the soft head plug technique were reliable (replicated) and, compared with results of the conventional method, valid. CONCLUSIONS: The results indicate that the soft head plug technique can be used to study relationships between neonatal sleep/wake variables and adult variables. PMID- 10678462 TI - The impact of split-night polysomnography for diagnosis and positive pressure therapy titration on treatment acceptance and adherence in sleep apnea/hypopnea. AB - STUDY OBJECTIVES: The time and resource intensive nature of the traditional two night paradigm for diagnosing and titrating positive pressure therapy for Obstructive Sleep Apnea/Hypopnea (OSA/H) contributes to patient care cost and limitation of service availability. Although split night polysomography (PSG(SN)) algorithms can establish a diagnosis of OSA/H and establish a positive pressure prescription for many patients, there has been only limited evidence that this strategy does not impair acceptance and adherence to treatment. The objective of this study was to test the null hypothesis that PSG(SN) does not adversely impact acceptance and adherence to positive pressure therapy for OSA/H compared with a standard two-night PSG strategy (PSG(TN)). DESIGN: Retrospective case-controlled study. SETTING: University-based sleep disorders program PATIENTS: Both PSG(SN) and PSG(TN) (control) patients were selected on the basis of having an initial medical/sleep evaluation by a full-time physician member of the University of Pittsburgh Sleep Disorders Program, must not have had prior diagnostic PSG or treatment for sleep-disordered breathing, and must have been followed by the Sleep Program team. Selection of PSG(SN) patients required the ability to be matched with a control patient. Both groups underwent evaluation during the same time period. Of 146 patients who underwent PSG(SN) between October 1995 and September 1997, 51 had their initial evaluation and subsequent follow-up by physician-staff members of our Program. Of these, 15 were excluded from analysis because of a previous diagnostic PSG's or prior OSA/H therapy. Also, matches were unavailable for 5 patients. Seven patients refusal to use positive pressure at home and were not available for assessment of adherence, but were included in analysis of therapeutic acceptance. Thus, analysis of the impact of PSG(SN) on adherence to positive pressure therapy was based on a data set of 24 patients in whom a PSG(SN) was performed and 24 patients who had PSG(TN). The two groups were matched for age, Apnea+Hypopnea Index (AHI) and gender. MEASUREMENT AND RESULTS: There were no significant differences between the PSG(SN) and PSG(TN) groups with respect to age, body mass index (BMI), Desaturation Event Frequency (DEF), Arousal Index (ArI) or the Epworth Sleepiness Score (ESS). The nadir of oxyhemoglobin saturation (SpO2) during sleep was lower in the PSG(TN) than PSG(SN) group (69.3+/-15 vs. 79.8+/-9, mean+/-SD, p=0.012). During positive pressure titration, the time spent at the final pressure which was prescribed for the patients were comparable in both groups (123.4+/-64 vs. 161+/-96 minutes, PSG(SN) and PSG(TN), respectively, p=0.17). Adherence to therapy was objectively assessed by the average daily run-time of the positive pressure device at the first meter-read following initiation of treatment (55.1+/-44 vs. 40.8+/-16 days following home set-up, PSG(SN) and PSG(TN), respectively, p=0.14). Depending whether or not patients with previous exposure to positive pressure therapy were included in the analysis, 84-86% of patients undergoing PSG(SN) accepted therapy. There was no difference between the groups with respect to adherence (5.1+/-4 vs. 4.6+/-3 hours, PSG(SN) and PSG(TN), respectively, p=0.64). CONCLUSIONS: In a population of predominantly moderate-to-severe OSA/H patients, PSG(SN) strategy does not adversely impact on adherence to positive pressure therapy over the first six weeks of treatment. Acceptance of therapy is comparable to that reported in the literature following PSG(TN). PMID- 10678463 TI - Prevalence and correlates of self-reported sleep problems among Chinese adolescents. AB - STUDY OBJECTIVES: This study examined the prevalence and correlates of sleep problems in Chinese adolescents. DESIGN AND SETTING: An epidemiological questionnaire survey was carried out in five high schools in Shandong Province of Mainland China. PARTICIPANTS: A total of 1365 adolescents between the ages of 12 and 18 years comprising 823 boys and 542 girls. MEASUREMENTS AND INTERVENTIONS: The participants completed a self-administered questionnaire regarding sleep duration, sleep problems, stressful life events, lifestyles, and personal and family characteristics. RESULTS: Mean sleep duration at night was 7.64 hours (SD = 0.86) and decreased with increasing age. Of the sample, 16.9% (95% CI = 13.2 20.5%) reported insomnia symptoms including difficulty initiating sleep (10.8%), difficulty maintaining sleep (6.3%), and early morning awakening (2.1%). Nightmares were reported more frequently by girls (chi2=20.09, p<0.001). Only 2.3% of the sample had ever taken hypnotic medication during the past month. Almost 22% of the subjects went to bed later than 12:00 PM at least once a week. Multiple logistic regression analysis showed that greater age, being at senior high school, doing no habitual physical exercise, poor physical health, self selection of diet, longer distance from home to school, and life stress experienced during the past 12 months were significantly associated with an increased risk of insomnia. CONCLUSIONS: Self-reported sleep problems in Chinese adolescents are common and associated with multiple factors. These findings suggest the need for comprehensive programs to prevent sleep problems in adolescents. PMID- 10678464 TI - An epidemiological study of insomnia among the Japanese general population. AB - The study examined the prevalence and correlates of insomnia in a representative sample (n=3030) from the general population of Japan. Using a structured questionnaire, we found that the overall prevalence of insomnia during the preceding month was 21.4%, including difficulty initiating sleep (DIS: 8.3%), difficulty maintaining sleep (DMS: 15.0%), and early morning awakening (EMA: 8.0%). Multiple logistic regression analysis showed that older age, being unemployed, lack of habitual exercise, poor perceived health, psychological stress, and being unable to cope with stress were associated with an increased prevalence of insomnia. These findings indicate that the prevalence of insomnia in the general population of Japan is comparable to that reported in Western countries, and that insomnia is associated with multiple psychosocial factors. PMID- 10678465 TI - Sleepiness at work among commercial truck drivers. AB - Two separate groups consisting of both long-haul (N=184) and short-haul (N=133) truck drivers were surveyed to examine the frequency of driver sleepiness-related problems at work during the previous three months and to assess the incidence of sleep apnea syndrome symptoms. We also aimed to identify factors likely to predict self-reported difficulties in staying alert in work driving, dozing off (sometimes referred to as microsleeps) at the wheel and near misses. The responses suggest that for approximately 13% of the long-haul drivers the mean driving time per shift exceeded the EEC regulation. About 40% of the long-haul drivers and 21% of the short-haul drivers reported having problems in staying alert on at least 20% of their drives. Over 20% of the long-haul drivers also reported having dozed off at least twice while driving. Near misses due to dozing off had occurred in 17% of these drivers. Factors indicating sleep apnea syndrome occurred in only about 4% of the long-haul drivers and in only two short-haul drivers. Work and individual related factors as well as factors indicating sleep apnea syndrome contributed only slightly to predicting driver sleepiness-related problems. This suggests that driver sleepiness-related problems tend to be shared by many of the professional drivers, rather than being a "specific" and permanent problem for a smaller portion of drivers. However, difficulties in sleep patterns, such as having difficulty falling asleep, were infrequent. PMID- 10678466 TI - Accuracy of oximetry with thermistor (OxiFlow) for diagnosis of obstructive sleep apnea and hypopnea. AB - OBJECTIVES: To evaluate the diagnostic accuracy for obstructive sleep apnea and hypopnea (OSAH) of the OxiFlow (OF) device which combines oximetry with recording of thermistor airflow. DESIGN & SETTING: Patients scheduled for overnight diagnostic polysomnography (PSG) were studied with OF either simultaneously during laboratory PSG (L-OF, n=86), at home on a separate night (H-OF, n=66), or both (n=55). PATIENTS: 97 patients with suspected OSAH, of whom 40 had OSAH defined as an apnea-hypopnea index (AHI) of more than 15 events per hour of sleep on PSG. INTERVENTIONS: NA. MEASUREMENTS & RESULTS: The automated respiratory disturbance index (RDI) generated by the OF software considerably underestimated the AHI by PSG for both L-OF and H-OF. Altering the parameters for hypopnea identification by the software did not improve this. Visual inspection of the computerized OF tracings added considerable diagnostic information, but a manual count of RDI during visual review overestimated AHI. For the identification of cases vs. non-cases of OSAH, receiver operating characteristic area-under-the curve statistics ranged from 0.77-0.90 for L-OF and from 0.71-0.77 for H-OF. Combining automated analysis with subsequent visual inspection of OF tracings yielded an overall sensitivity of 86% and specificity of 74% for the diagnosis of OSAH during H-OF recordings. Analysis of potential technician time saved indicated a benefit from the use of OF. CONCLUSIONS: OF has diagnostic utility for the identification of OSAH. However, because of hardware and software limitations, it is unclear whether this device is superior to oximetry alone. PMID- 10678467 TI - Sleep latency and duration estimates among sleep disorder patients: variability as a function of sleep disorder diagnosis, sleep history, and psychological characteristics. AB - STUDY OBJECTIVES: Insomnia patients often report greater sleep disturbance than found via polysomnography; yet the specific patient factors related to such sleep time misperceptions are poorly understood. We sought to characterize the extent to which a diverse group of patients complaining of insomnia (n=104) misperceive overnight total sleep time and sleep latency, and to identify patient factors associated with these variations. DESIGN: Cross-sectional. SETTING: University based sleep disorders center. PATIENTS: Sleep disorder groups consisted of patients with psychophysiological insomnia (n=19), sleep state misperception (n=8), insomnia with depressive disorder (n=11), insomnia secondary to Axis I psychiatric disorder other than depression (n=21), periodic limb movement disorder (n=24), and obstructive sleep apnea (n=21). MEASUREMENT AND RESULTS: Patients completed a sleep history questionnaire and the MMPI, underwent overnight diagnostic polysomnographic assessment, and then estimated their total sleep time and sleep latency the subsequent morning. On average, patients overestimated sleep latency, but were equally likely to underestimate vs. overestimate total sleep time. Sleep time misperception was associated with longer periods of wakefulness following sleep onset, greater self-perceived sleep impairment, as well as several psychological dimensions. CONCLUSIONS: Patient factors, including sleep quality, perceptions of habitual sleep time, and current psychopathology, potentially influence sleep time estimation. Whereas psychological factors may lead to exaggeration of sleep disturbance among some patients, sleep quality itself may also influence the congruence between subjective and objective indices of sleep. PMID- 10678468 TI - Sleep deprivation and phasic activity of REM sleep: independence of middle-ear muscle activity from rapid eye movements. AB - In the recovery nights after total and partial sleep deprivation there is a reduction of rapid eye movements during REM sleep as compared to baseline nights; recent evidence provided by a selective SWS deprivation study also shows that the highest percentage of variance of this reduction is explained by SWS rebound. The present study assesses whether the reduction of rapid eye movements (REMs) during the recovery night after total sleep deprivation is paralleled by a decrease of middle-ear muscle activity (MEMA), another phasic muscle activity of REM sleep. Standard polysomnography, MEMA and REMs of nine subjects were recorded for three nights (one adaptation, one baseline, one recovery); baseline and recovery night were separated by a period of 40 hours of continuous wake. Results show that, in the recovery night, sleep deprivation was effective in determining an increase of SWS amount and of the sleep efficiency index, and a decrease of stage 1, stage 2, intra-sleep wake, and NREM latencies, without affecting REM duration and latency. However, MEMA frequency during REM sleep did not diminish during these nights as compared to baseline ones, while there was a clear effect of REM frequency reduction. Results indicate an independence of phasic events of REM sleep, suggesting that the inverse relation between recovery sleep after sleep deprivation and REM frequency is not paralleled by a concomitant variation in MEMA frequency. PMID- 10678469 TI - Daily social and physical activity increases slow-wave sleep and daytime neuropsychological performance in the elderly. AB - Decreased levels of physical and social activity associated with aging can be particularly pronounced in residents of assisted living facilities. Reduced exposure to important behavioral and time-giving cues may contribute to the age related changes in circadian rhythmicity and sleep. The present study was conducted to test the hypothesis that an enforced schedule of structured social and physical activity (0:900 to 10:30 and 19:00 to 20:30 daily for two weeks) can have beneficial effects on circadian rhythmicity, nocturnal sleep, daytime functioning, mood, and vigor. The subjects were 14 elderly residents of continued care retirement facilities while a similar group of 9 elderly residents served as controls. The group exposed to structured activities had increased amounts of slow-wave sleep and demonstrated improvement in memory-oriented tasks following the intervention. Conversely, no significant changes were noted in the amplitude and phase of the body temperature rhythm or in subjective measures of vigor and mood. These results indicate that short-term exposure to structured social intervention and light physical activity can significantly improve memory performance and enhance slow-wave sleep in older adults without alterations to the circadian phase or amplitude of body temperature. This is the first report to demonstrate that low intensity activity in an elderly population can increase deep sleep and improve memory functioning. The high degree of interest in these activities paired with the simple nature of the tasks makes this a potentially practical intervention which can be adapted for both community dwelling and assisted-living elders. PMID- 10678471 TI - Recent literature in sleep research. PMID- 10678470 TI - The roles of vertex sharp waves and K-complexes in the generation of N300 in auditory and respiratory-related evoked potentials during early stage 2 NREM sleep. AB - STUDY OBJECTIVES: To determine the scalp topography of the N300 response to stimuli of different modalities and to investigate the relationship of the N300 component to K-complexes and vertex sharp waves seen in the un-averaged EEG. DESIGN: Two experiments were conducted one using auditory; the other using respiratory occlusion stimuli presented during stage 2 sleep. Trials were classified on the basis of whether they produced a K-complex, a vertex sharp wave, or some other response. Auditory stimuli were presented in the form of an oddball paradigm, and averaged separately depending on whether they were "frequent" or "rare". In both experiments, responses were averaged separately based on the appearance of K-complexes, vertex sharps waves, or some "other" response to the stimuli. SETTING: Data were collected in the Melbourne University Sleep Laboratory. PARTICIPANTS: Young healthy male adults, eight in experiment 1 and six in experiment 2. INTERVENTIONS: NA. MEASUREMENTS AND RESULTS: Data were collected from 29 scalp sites. In all cases, N300 amplitude was maximal in the vertex sharp wave averages, despite being clearly present in the averages of K complexes and "other" responses. The vertex maximal scalp topography of the N300 did not differ across response conditions or as a function of stimulus modality. This is consistent with the N300 being produced by the same intracranial generators in all cases. There were no effects of stimulus or response type on N300 latency. CONCLUSIONS: N300 should be viewed as a multi-modal component with a different underlying generator mechanism than that of the K-complex. PMID- 10678472 TI - The emerging AIDS crisis in Russia: review of enabling factors and prevention needs. AB - Eastern Europe is experiencing increased rates of HIV/AIDS, and the Russian Federation is among the countries with the most alarming case rate increases. Behavioural and biological studies demonstrate that the transmission of HIV in Russia is occurring as a result of injection drug use, homosexual, and heterosexual risk behaviours. Factors that promote risk and therefore enable HIV transmission in Russia parallel those found in other countries, including epidemics of other sexually transmitted infections, economic instability, poverty, and social factors such as gender roles. Research is urgently needed to better understand and forecast the HIV epidemic in Russia, as well as to develop effective interventions to prevent a Russian AIDS crisis. PMID- 10678473 TI - Mycoplasma species in rapid and slow HIV progressors. AB - We determined the relationship between the presence of Mycoplasma fermentans and Mycoplasma penetrans and the rate of progression of HIV-associated disease in a nested case-control study based on a cohort of 159 HIV-infected patients with different rates of disease progression. Study participants were divided into 3 progression groups: non-progressors who had been HIV-1 seropositive for at least 9 years and had remained asymptomatic with a CD4 cell count of > 500/mm3; slow progressors who had been HIV-1 seropositive for at least 9 years and whose CD4 cell count had fallen below 500 cells, and who had developed symptomatic disease or AIDS; and rapid progressors who had developed AIDS within 5 years of HIV infection. Peripheral blood mononuclear cells (PBMCs) were collected at enrollment and examined by mycoplasma polymerase chain reaction (PCR) assays. Three (7%) of 46 non-progressors, 3 (3%) of 86 slow progressors, and 2 (7%) of 27 rapid progressors were M. fermentans positive. The PBMCs from 91 subjects were tested for M. penetrans DNA and none was positive. The small proportion of M. fermentans-positive patients indicates that the mycoplasma cannot be important in the development of AIDS in the large majority of patients. Furthermore, no association was found between its presence and more rapid HIV disease progression. PMID- 10678474 TI - Asymptomatic penile HPV infection: a prospective study. AB - The occurrence of human papillomavirus (HPV) among males was analysed with the polymerase chain reaction (PCR) method. Penile brush samples were taken once from 147 males attending for a control or for HPV non-related reasons, and consecutive samples were collected from 88 males re-attending the clinic. Of the males attending once, 13% (19/147) were HPV DNA positive and among the re-attenders 14% (12/88) were initially positive as compared with 33% (29/88) who were positive at least at one visit. Totally, 22 different HPV types were detected of which HPV 16 was most common, found in 6.4% (15/235), followed by HPV 42 found in 3.8% (9/235). Among 14 HPV-positive males with at least one follow-up, 7 had persistent infections with at least one HPV type, and transient HPV types were observed in 9; but in 5 of them new types appeared at follow-up. Among sexually active males subclinical/latent HPV infection is common and repeated sampling increases its prevalence. PMID- 10678475 TI - Detection of human cytomegalovirus retinitis and monitoring of ganciclovir treatment using conjunctival swab with polymerase chain reaction in AIDS patients. AB - This report studies the accuracy of conjunctival swab polymerase chain reaction (CS-PCR) for the diagnosis of human cytomegalovirus retinitis (HCMV) in AIDS patients. PCR and virus culture were used for the detection of HCMV in conjunctival swab, serum, and urine specimens from 38 AIDS patients between April 1996 and April 1998. The clinical utility of the identification of HCMV retinitis by these 6 different methods was demonstrated by their prediction power to estimate AIDS patients at risk of contracting HCMV retinitis. The sensitivity, specificity, positive predictive value, and negative predictive value of CS-PCR for the detection of HCMV retinitis were 91.5%, 80.9%, 60.8%, and 92.7%, respectively; for serum PCR were 74.3%, 81.7%, 57.2%, and 90.3%; for urine PCR were 100%, 17.3%, 20.4%, and 100%; for conjunctival swab culture were 22.7%, 100%, 100%, and 86%; for serum culture were 27.3%, 98.1%, 75%, and 86.4%; and for urine culture were 90.9%, 44.2%, 25.6%, and 95.8%. PMID- 10678476 TI - A survey of patients with Chlamydia trachomatis infection: sexual behaviour and perceptions about contact tracing. AB - The aim of this study was to evaluate how patients with Chlamydia trachomatis infection perceived the legal enforcement of partner notification and to seek their views on legislation impinging on their own sexual behaviour. The investigation was performed at STD clinics in Stockholm, Sweden in 1997. Consecutive patients (n=192) answered a questionnaire about sexual behaviour and contact tracing. More men (40%) than women (21%) had had sexual intercourse during the past 6 months with an occasional partner. The mean number (6 months prior to this) was 2.3 partners (1-15) for men and 2.2 partners (1-21) for women. Eighteen per cent admitted to having avoided disclosing the name of their partner(s). Ninety per cent considered it beneficial that chlamydial infection was regulated and that a named partner could be forced to undergo STD testing. Partly based on this report, the government has recommended police enforcement to be removed from the legislation as a tool for contact tracing in chlamydial infections. PMID- 10678477 TI - Condom use and 'withdrawal': exploring gay men's practice of anal intercourse. AB - Some gay men who have unprotected anal intercourse avoid ejaculation-they practise 'withdrawal'. Using data collected in 1997 from a sample of Sydney gay men (n=625), we explored the relation between men's practice of ejaculation and their use of condoms. We also investigated whether men who had unprotected withdrawal but not unprotected ejaculation were more likely to think unprotected withdrawal was safe, liked condoms less, liked anal intercourse more, or were more sexually adventurous. Considering separately insertive and receptive anal intercourse with regular and with casual partners, we found that the majority of men who practised unprotected withdrawal also practised unprotected ejaculation. Of those whose only unprotected sex was withdrawal ('true withdrawers'), most never used condoms (they did not also have protected sex with ejaculation). True withdrawers were compared with men who had unprotected ejaculation, who always used condoms, who had no anal sex and who had no partners. Those who were true withdrawers with casual partners were more likely to believe withdrawal was safe; no group effects were found with regular partners. No significant differences in condom attitudes were found. True withdrawers with regular partners liked anal intercourse less than other men, but true withdrawers with casual partners were indistinguishable from those who had unprotected ejaculation. True withdrawers did not differ in sexual adventurousness from other men who had anal intercourse. Most withdrawers avoided anal sex with ejaculation rather than use condoms. Converting them into reliable condom users may be a considerable challenge for health promotion. PMID- 10678478 TI - Evaluation of a drama-in-education programme to increase AIDS awareness in South African high schools: a randomized community intervention trial. AB - A community intervention trial was undertaken in KwaZulu Natal, South Africa to evaluate the effectiveness of a high school drama-in-education programme. Seven pairs of secondary schools were randomized to receive either written information about HIV/AIDS or the drama programme. Questionnaire surveys of knowledge, attitude and behaviour were compared before and 6 months after the interventions. One thousand and eighty students participated in the first survey and 699 in the second. Improvements in knowledge (P=0.0002) and attitudes (P < 0.00001) about HIV/AIDS were demonstrated in pupils at schools receiving the drama programme when compared to pupils receiving written information alone. These changes were independent of age, gender, school or previous sexual experience. In schools receiving the drama programme, sexually active pupils reported an increase in condom use (P < 0.01). It is important to provide resources to sustain such programmes and to obtain stronger evidence of effect on behaviour by measuring changes in HIV incidence. PMID- 10678479 TI - Risk perception and counselling among HIV-positive women in Sao Paulo, Brazil. AB - This study of HIV-positive women at a clinic for HIV/AIDS in Sao Paulo examined their risk perception for HIV before they had learned of their diagnosis and their experiences with pre- and post-test counselling. A sample of 148 women was interviewed regarding demographics, HIV risk factors and risk perception, pre- and post-test counselling, and sexual and reproductive conduct. The majority (77%) had been infected by their partners--37% by an injecting drug user partner. More than half (53%) did not perceive themselves at risk before learning of their HIV status and, of 68 who had perceived themselves at risk, 29% did so only after their partners became ill. The majority (64%) did not receive any kind of pre test counselling. Post-test counselling was reported by 83% but 14% reported being mishandled by a physician in the process. Findings suggest the importance of prevention efforts to reduce women's barriers to learning about HIV status and the necessity of improving the quality of pre- and post-test counselling. PMID- 10678480 TI - Trend of antimicrobial resistance in Neisseria gonorrhoeae at New Delhi, India. AB - We aim to monitor the trends of antimicrobial resistance in Neisseria gonorrhoeae and to compare the results of antimicrobial sensitivity by disc diffusion and minimum inhibitory concentration (MIC). Two hundred and eleven confirmed strains of N. gonorrhoeae were subjected to antimicrobial sensitivity testing by disc diffusion using penicillin, tetracycline, ciprofloxacin and ceftriaxone from 1995 to June 1999. Penicillinase-producing Neisseria gonorrhoeae (PPNG) were detected by lodometric method. Minimum inhibitory concentration was determined by E test. A low level of penicillin resistance and PPNG detected in 1996 was maintained over the years. Significant increasing trend of tetracycline and ciprofloxacin resistance with high MIC i.e. 2-96 microg/ml and 1-32 microg/ml respectively were found. Ceftriaxone was found to be the drug of choice, being 100% sensitive. Comparison of resistance pattern by the 2 tests showed satisfactory agreement. Emergence of penicillin, quinolone and tetracyline resistance in N. gonorrhoeae isolates from a major STD centre at New Delhi indicates the need for increased awareness, prudent use of antimicrobials, and evaluation of new antimicrobials for the treatment of gonorrhoea. PMID- 10678481 TI - Lymphocytic interstitial pneumonitis in an HIV-infected adult: response to antiretroviral therapy. PMID- 10678482 TI - An annular erythema secondary to primary genital herpes. PMID- 10678483 TI - Disseminated Penicillium marneffei in a patient infected with human immunodeficiency virus. AB - A case of a 31-year-old man with systemic Penicillium marneffei infection acquired in Thailand and who developed endophthalmitis is described. This presentation has not previously been reported. He responded to combined treatment with intravenous and intravitreal amphotericin. PMID- 10678484 TI - Sexual health knowledge of adolescents in a Great Yarmouth high school. PMID- 10678485 TI - The need to exercise caution in the management of patients co-infected with HIV and hepatitis B. PMID- 10678486 TI - Four-drug salvage antiretroviral treatment including nelfinavir and efavirenz in highly treated children with congenital HIV disease. PMID- 10678487 TI - Behavioural and biomedical risk factors for the transmission of HIV/AIDS in Bangladesh. PMID- 10678488 TI - Evaluation and prediction of drug permeation. AB - A major challenge confronting the pharmaceutical scientist is to optimize the selective and efficient delivery of new active entities and drug candidates. Successful drug development requires not only optimization of specific and potent pharmacodynamic activity, but also efficient delivery to the target site. Following advances in rational drug design, combinatorial chemistry and high throughput screening techniques, the number of newly discovered and promising active compounds has increased dramatically in recent years, often making delivery problems the rate-limiting step in drug research. To overcome these problems, a good knowledge of the pharmacokinetic barriers encountered by bioactive compounds is required. This review gives an overview of the properties of relevant physiological barriers and presents some important biological models for evaluation of drug permeation and transport. Physicochemical determinants in drug permeation and the relevance of quantitative and qualitative approaches to the prediction and evaluation of passive drug absorption are also discussed. PMID- 10678489 TI - Biochemical and pharmacological characteristics of a newly synthesized H+/K+ ATPase inhibitor, YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7 naphthyridine, in pigs and rats. AB - We have investigated the effect of the newly synthesized proton-pump inhibitor YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, on gastric mucosal proton pump (H+/K+-ATPase) activity, gastric acid secretion and gastric lesions in experimental animals. In lyophilized pig gastric microsomes, YJA20379-8 was shown to inhibit H+/K+-ATPase activity; the inhibitory effect was not affected by pH, the IC50 (dose resulting in 50% inhibition) being 28.0 microM and 30.0 microM at pH 6.4 and pH 7.4, respectively. The effect was fully reversed by dilution and subsequent washing of the incubation mixtures of H+/K+-ATPase and YJA20379-8, suggesting the reversible nature of the enzyme inhibition. In pylorus ligated rats, YJA20379-8 administered by different routes (intraduodenal, subcutaneous, intravenous or oral) resulted in dose-dependent suppression of basal gastric acid secretion. The duration of antisecretory action of 30 mg kg( 1) YJA20379-8 given intraduodenally was very brief (less than 7 h). Pretreatment with YJA20379-8 also dose-dependently prevented gastric lesions induced by absolute ethanol and water-immersion stress in rats. These results suggest that YJA20379-8 might exert its antiulcer activity partly by reversible suppression of acid secretion and partly by protecting the gastric mucosa against ulcerative stimuli. PMID- 10678490 TI - Inclusion complexation of the sunscreen agent 2-ethylhexyl-p dimethylaminobenzoate with hydroxypropyl-beta-cyclodextrin: effect on photostability. AB - The interaction between the UV filter, 2-ethylhexyl-p-dimethylaminobenzoate, and unmodified and modified alpha-, beta- or gamma-cyclodextrins was studied in water by phase-solubility analysis. Of the cyclodextrins available, only hydroxypropyl beta-cyclodextrin caused a marked increase in the aqueous solubility of 2 ethylhexyl-p-dimethylaminobenzoate. The data from the solubility study indicated the formation of a 1:1 (sunscreen-cyclodextrin) complex. The inclusion of the sunscreen agent into the hydroxypropyl-beta-cyclodextrin cavity was confirmed by thermal analysis and by nuclear magnetic resonance spectroscopy. Irradiation induced degradation of 2-ethylhexyl-p-dimethylaminobenzoate was reduced by complexation with hydroxypropyl-beta-cyclodextrin, this effect being more pronounced in solution (the extent of degradation was 25.5% for the complex compared with 54.6% for free 2-ethylhexyl-p-dimethylaminobenzoate) than in the emulsion vehicle (the extent of degradation was 25.1% for the complex compared with 33.4% for free 2-ethylhexyl-p-dimethylaminobenzoate). Although photodegradation of the sunscreen agent is significantly reduced by formation of the inclusion complex it is important to design a suitable vehicle. Inclusion of 2-ethylhexyl-p-dimethylaminobenzoate-DMAB into the hydroxypropyl-beta cyclodextrin cavity limits interaction of the UV filter with the skin reducing the side-effects of the formulation. PMID- 10678491 TI - Absorption of vitamin K2 by dogs after oral administration of a soft gelatin capsule formulation containing a new emulsion-type vehicle. AB - This study has evaluated the performance of a newly developed vehicle for administration of a drug in a soft gelatin capsule. The absorption of vitamin K2 in dogs after oral administration of the vitamin in a soft gelatin capsule containing the newly developed vehicle was compared with absorption after administration of a control formulation prepared by encapsulating the contents of a commercially available vitamin K2 capsule (Glakay capsules 15 mg) in the same type of soft gelatin. Under non-fasted conditions the profile of the plasma concentration of vitamin K2 against time for the test formulation was comparable with that for the control formulation in non-fasted dogs. Under fasted conditions, however, both the maximum concentration (Cmax) and the area under the plot of concentration against time (AUC) were significantly smaller for the test formulation than for the control formulation. The Cmax and AUC for the test formulation were about 10 times larger for non-fasted dogs than for fasted dogs whereas values for the control formulation were about twice as large. These results suggest that both formulations might require the presence of food or digestive fluid components, or both, for better absorption of vitamin K2. It seems that although the performances of the test and control formulations were comparable in the presence of these components, the control formulation works better in their absence. It should be also noted that, in contrast with the results from the absorption tests, the dispersibility of the test vehicle in water was much better than that of the control vehicle. This suggests that dispersibility does not significantly affect vitamin K2 absorption. In conclusion, although the new vehicle did not perform better than the control vehicle in terms of vitamin K2 absorption, the performance of the control formulation was comparable for non-fasted dogs. Because the new vehicle contains considerably less surfactant than the vehicles currently used in soft gelatin capsules, it could be a safer alternative for use under non-fasted conditions. PMID- 10678492 TI - Statistical moments for placental transfer of solutes in man. AB - The placental transfer of red blood cells and solutes in man has been investigated by statistical moment analysis, using the impulse-response technique. Model compounds of different lipophilicity (sucrose, water, antipyrine, propranolol and labetalol) were injected with a vascular reference (labelled red blood cells) as boluses into either the foetal or maternal circulation of a single-pass perfused placental lobule. Maternal and foetal venous outflow fractions were collected at intervals ranging from 1 to 600 s. Perfusion was conducted at maternal flow rates of 4 and 6 mL min(-1) and foetal flow rates of 2 and 3 mL min(-1), respectively, to yield a constant materno foetal flow ratio of 2. The outflow concentration-time profile curves were analysed by statistical moment analysis. The sum of foetal and maternal recovery was close to 100% for red blood cells, sucrose, water and antipyrine, but lower for propranolol and labetalol. The mean transit time (MTT) values ranged from 20 to 500 s. The normalized variance (CV2) for red blood cells in the foetal and maternal circulation of the placenta were in the ranges 2.31 to 3.86 and 2.00 to 2.03, respectively. The shape of the outflow concentration-time profiles after bolus input is consistent with that of vascular residence time models such as the dispersion model. The heterogeneity in red blood cell transit times, as defined by CV2, is greater than in either the perfused leg or perfused liver. PMID- 10678493 TI - Effect of oral administration of fennel (Foeniculum vulgare) on ciprofloxacin absorption and disposition in the rat. AB - The aim of this study was to investigate the possibility of a drug-drug interaction between ciprofloxacin and fennel (Foeniculum vulgare) in a rat model. Pharmacokinetic assessment of ciprofloxacin was performed in two groups of male Sprague-Dawley rats. One group (n = 5) received 20 mg kg(-1) antibiotic orally with concomitant oral dosing of the aqueous fennel extract (2 g herb kg(-1)) whereas the controls (n = 5) received 20 mg kg(-1) oral ciprofloxacin. Blood and urine samples were collected over 6 and 24 h, respectively, for quantitation of ciprofloxacin by HPLC. A non-compartmental model was employed for pharmacokinetic analysis. Major ingredients and metal cations in the fennel extract were determined. Compared with the control, maximum plasma concentration, area under the curve and urinary recovery of ciprofloxacin were significantly (P < 0.05) lower, by 83, 48 and 43%, respectively, in rats receiving concomitant dosing of the two agents. The relative bioavailability of ciprofloxacin, under the influence of fennel, was estimated to be 0.52. In addition, its apparent volume of distribution and terminal elimination half-life were significantly (P < 0.05) increased, from 30.8 +/- 11.1 (L kg(-1)) and 2.0 +/- 0.4 (h) to 143.8 +/- 31.6 (L kg(-1)) and 5.2 +/- 2.0 (h), respectively. Although none of the organic components of fennel seemed to cause this interaction, the total amount of ten metal cations measured was found to be 13 mg g(-1). Significant interaction between ciprofloxacin and fennel was observed in this study. Absorption, distribution and elimination of ciprofloxacin were all affected. These changes might be because of the formation of a more lipophilic ciprofloxacin chelate in the presence of relatively large amounts of metal cations. If, therefore, the two therapeutic agents are used concurrently, an adequate dosing interval is needed to ensure the efficacy of ciprofloxacin. PMID- 10678494 TI - The mechanisms of immune suppression by high-pressure stress in mice. AB - The effects of high-pressure stress on the induction of anti-sheep red blood cells (SRBC) and of plaque-forming cells (PFC), and on thymus weight, were studied in BALB/c mice in-vivo and in-vitro. The efficacy of high-pressure stress in suppressing PFC and thymic involution was maximum when the stress was applied 1 h day(-1) for 2 days before immunization with SRBC. Both effects were blocked by administration of indomethacin, atropine, naloxone or phentolamine before the first application of stress, whereas hexamethonium and propranolol had no such effect. Hexamethonium, naloxone and propranolol administered before the second application of high-pressure stress blocked both effects. Prostaglandin and acetylcholine given 24 h before application of high-pressure stress caused a marked reduction in PFC count, but not in thymus weight. The reduced PFC count caused by acetylcholine was blocked by pretreatment with indomethacin. When adrenaline was injected 24 h after application of high-pressure stress a marked reduction in PFC was observed, but without thymic involution. When adrenaline was injected 24 h after prostaglandin injection the PFC count was also markedly reduced, but not thymus weight. The decrease in PFC caused by two exposures to stress or one exposure to stress plus injection of adrenaline was blocked by diethylcarbamazine before the second exposure to stress or the injection of adrenaline. In addition, normal spleen cells, were induced as suppressor cells when incubated with the serum of stressed mice, but not when supplemented with anti-leukotriene C4, D4 antibody. These data suggest that mice fall into a pre stress condition via the release of prostaglandin after the first stress, and then immunosuppression is induced in these prestressed mice via the release of leukotriene C4, D4, caused by the activation of the autonomic nervous system by the second exposure to stress. PMID- 10678495 TI - Time-dependent striatal dopamine depletion after injection of 6-hydroxydopamine in the rat. Comparison of single bilateral and double bilateral lesions. AB - For future investigation of possible perturbation of circadian rhythm in animal models of Parkinson's disease we needed an animal model providing lasting 80-100% striatal dopaminergic depletion in rats, but without induced mortality. We have thus compared the effects of a single hydroxydopamine bilateral striatal lesion (SB-hydroxydopamine) with those of a double hydroxydopamine bilateral lesion (DB hydroxydopamine) at the same dose (16 microg/striatum) by HPLC determination of dopamine and 3,4-dihydrophenylacetic acid (dopac) levels in the striatum. Two weeks after neurosurgery, SB-hydroxydopamine and DB-hydroxydopamine induced dopaminergic depletion of at least 81% compared with control groups. After eight weeks striatal dopaminergic depletion was only 60.97% in SB-hydroxydopamine rats, suggesting a compensatory phenomenon, whereas in DB-hydroxydopamine rats dopaminergic loss was stable at 88%. For the DB-hydroxydopamine group the dopac/dopamine ratio was significantly increased at week 2 only, whereas no significant change was observed for other groups. This increase might be explained by increased dopamine turnover. We have demonstrated that striatal DB hydroxydopamine injection induces lasting 80-100% neuronal loss, close to that observed in the disease in man, without induced mortality, and provides a tool which meets our experimental requirements. PMID- 10678496 TI - Oestrogenic compounds modulate cytokine-induced nitric oxide production in mouse osteoblast-like cells. AB - Nitric oxide (NO) is a mediator of bone metabolism with effects on both bone resorption and formation. Its production by both the constitutive and inducible isoforms of nitric oxide synthase (NOS) is affected by oestrogen in several types of cell and in tissues other than bone cells. Recently, oestrogens were found to increase basal NO production by osteoblasts via enhanced activity or expression, or both, of NOS-3. Inflammatory cytokines, however, increase NO by increasing the expression of NOS-2. In this study we have examined whether cytokine-induced NO production by osteoblastic cells was affected by oestrogenic compounds by studying the effect of 17beta-oestradiol and the anti-oestrogens ICI164,384 and 4 hydroxytamoxifen on cytokine-induced NO production in oestrogen receptor positive MC3T3-E1 osteoblast-like cells. Combinations of the inflammatory cytokines interleukin-1beta, tumour necrosis factor-alpha, and interferon-gamma with lipopolysaccharide stimulated NO production up to 11-fold. This cytokine-induced NO production was further increased dose-dependently by the anti-oestrogens ICI164,384 and 4-hydroxytamoxifen (133.3 +/- 3.2% and 146.0 +/- 13.2%, respectively). 17Beta-oestradiol either had no effect on or slightly inhibited cytokine-induced NO production. It did, however, dose-dependently counteract the stimulatory effect of the anti-oestrogens. Concentrations of 17beta-oestradiol needed to prevent the stimulatory effect of 4-hydroxytamoxifen were ca tenfold that of ICI164,384. These findings show that, in addition to the stimulatory effect of oestrogen on basal NO production by NOS-3, cytokine-induced NO production is also affected by oestrogenic compounds in osteoblasts. PMID- 10678497 TI - Effects of 5-hydroxytryptamine on the transintestinal electrical activity and cardiovascular function of fawn-hooded rats in-vivo. AB - Fawn-hooded rats, which have abnormal serotonergic function, were used to investigate the receptors involved in 5-hydroxytryptamine (5-HT)-induced intestinal secretion. The effects of 5-HT on secretion by the small intestine and proximal colon, monitored as increased transintestinal electrical activity, and on cardiovascular function, measured as changes in heart rate and blood pressure, were compared in fawn-hooded and Wistar rats. The maximum fall in heart rate induced by 5-HT (mediated by 5-HT3 receptors) was greater in fawn-hooded than in Wistar rats. ED50 values (the doses resulting in 50% of the maximum effect) for the 5-HT2-mediated increases in systolic pressure were lower for both 5-HT and 5 methoxytryptamine in the fawn-hooded group. The prolonged fall in diastolic pressure mediated by 5-HT1-like receptors was significantly attenuated in fawn hooded rats, with the maximum responses to 5-HT, 5-methoxytryptamine and 6 hydroxyindalpine reduced to 21%, 42% and 28%, respectively, of the values obtained for Wistar rats. In fawn-hooded rats the small intestine was less sensitive to the effects of 5-HT (ED50 = 47 nmol kg(-1); ED50 for Wistar rats = 23 nmol kg(-1)) and the maximum colonic response to 5-methoxytryptamine was greater (7.0 mV compared with 4.3 mV in Wistar rats), but other indices did not differ for the two strains. The responses to 6-hydroxyindalpine were similar in fawn-hooded and Wistar rats. It is concluded that although the cardiovascular response of fawn-hooded rats to 5-HT challenge is very different from that of Wistar rats, this difference is not reflected in marked alterations in 5-HT induced intestinal secretion. This is consistent with 5-HT stimulating secretion via the activation of several different receptor subtypes so that any changes in the receptor profile in fawn-hooded rats results in little alteration in the overall intestinal response. PMID- 10678498 TI - Effect of pretreatment with FTY720 and cyclosporin on ischaemia-reperfusion injury of the liver in rats. AB - The effect of pretreatment with FTY720 (2-amino-2-[2-(4-octylphenyl)ethyl]-1,3 propanediol hydrochloride) or cyclosporin, or both, on neutrophil-mediated injury has been examined by use of a rat model of transient clamping of hepatic flow. Pretreatment with FTY720 alone or with cyclosporin induced a marked reduction of circulatory lymphocytes, whereas the use of these drugs in combination was very effective at suppressing the elevation of the number of peripheral polymorphonuclear neutrophils (PMN) after reperfusion. Pretreatment with cyclosporin, with or without FTY720, significantly reduced hepatic damage, whereas FTY720 alone tended to prolong hepatic damage. Pretreatment of cyclosporin alone, but not in combination with FTY720, significantly reduced the accumulation of PMN and led to lower myeloperoxidase levels in the damaged liver. In conclusion, pretreatment with cyclosporin, with or without FTY720, reduced hepatic damage after warm ischaemia-reperfusion, whereas pretreatment with FTY720 alone tended to prolong this damage. PMID- 10678499 TI - Modulation of nitric oxide-dependent vascular and platelet function in-vitro by the novel phosphodiesterase type-V inhibitor, ONO-1505. AB - We have characterized the in-vitro modulation of both nitric oxide (NO)-dependent vasodilator activity and anti-platelet function by the novel type-V phosphodiesterase inhibitor, ONO-1505 (4-[2-(2-hydroxyethoxy)ethylamino]-2-(1H imidazol-1-yl)-6-methoxyquin azoline methanesulphonate). ONO-1505 elicited vasorelaxation in the rat isolated aorta. If the concentration of ONO-1505 was < or = 10 microM the vasorelaxation was abolished by N(G)-nitro-L-arginine methyl ester (L-NAME), by methylene blue, and by endothelial denudation. Furthermore, pretreatment of the rat isolated aorta for 10 min with ONO-1505 in the presence of L-NAME potentiated vasorelaxation to the NO-donor, sodium nitroprusside. Similarly, ONO-1505, although having no effect on adenosine diphosphate (ADP) induced rat platelet aggregation in-vitro, augmented established anti-aggregatory effects of sodium nitroprusside. The data therefore show that the novel phosphodiesterase V inhibitor ONO-1505 augments endogenous and exogenous nitrovasodilator activity in-vitro; they also imply modulation of the NO pathway in the haemodynamic actions of this compound, previously reported in-vivo. PMID- 10678500 TI - In-vivo and in-vitro assessment of the free-radical-scavenger activity of Ginkgo flavone glycosides at high concentration. AB - Free radicals are involved in numerous skin diseases, especially inflammatory reactions and photosenescence. To identify possible free-radical scavenging by an original terpene-free Ginkgo biloba extract containing 33% Ginkgo flavone glycosides, mostly quercetin and kaempferol derivatives, we studied its activity by means of in-vitro and in-vivo experiments, using superoxide dismutase (SOD) as a positive control. By means of an in-vitro electron-spin resonance (ESR) assay we compared the activity of the Ginkgo extract with that of its two aglycones, quercetin and kaempferol. Quercetin and Ginkgo extract had significant antioxidant properties without pro-oxidant effect. In contrast, kaempferol, above an optimum antioxidant concentration, behaved as a pro-oxidant. The in-vivo experiments were conducted on an anti-inflammatory model. The cutaneous blood flux which reflects the skin inflammatory level was recorded by means of a laser Doppler perfusion imager. The data confirmed the free-radical-scavenging property of both Ginkgo extract and SOD. The Ginkgo extract significantly inhibited (37%) cutaneous blood flux to the same extent as SOD. These data confirmed the antioxidant property of Ginkgo extract. A complementary spin-trapping technique would enable identification of the free radicals involved. This Ginkgo extract should be useful for protection of the skin against free radicals. PMID- 10678501 TI - Isolation of opioid-active compounds from Tabernaemontana pachysiphon leaves. AB - A procedure for prefractionation of crude plant extracts by centrifugal partition chromatography (CPC) has been developed to enable rapid identification of known positive compounds or false-positive compounds and to increase the chance of identifying minor unknown-active compounds. The study explored the use of CPC as a tool in the prefractionation step before investigation of bioactivity. Fractions obtained by CPC from an ethanolic extract of Tabernaemontana pachysiphon Stapf (Apocynaceae) were screened by means of an opiate-receptor binding assay and an adenosine A1-receptor-binding assay. Fractions containing fatty acids, which had false-positive effects on the assay, were identified, as were unknown-positive fractions from which two opioid-active compounds, tubotaiwine and apparicine, were subsequently isolated. The affinities (Ki) of tubotaiwine and apparicine at the opiate receptor were 1.65 +/- 0.81 and 2.65 +/- 1.56 micromol, respectively. Both alkaloids had analgesic activity in the abdominal constriction test in mice. CPC prefractionation led to the rapid isolation of two opioid-active compounds, tubotaiwine and apparicine, from the unknown-positive fraction; false-positive fractions were rapidly identified. Both tubotaiwine and apparicine had affinity for adenosine receptors in the micromolar range and also had in-vivo analgesic activity in mice. PMID- 10678502 TI - Anti-inflammatory and antinociceptive effects in rodents of the essential oil of Croton cajucara Benth. AB - The plant Croton cajucara Benth. (Euphorbiaceae) is widely used in Amazonian folk medicine for the treatment of a wide range of illnesses. In this investigation the analgesic and anti-inflammatory properties of the essential oil from the bark of C. cajucara Benth., administered orally, were determined in several standard rodent models of pain and inflammation. We observed that pretreatment with essential oil significantly reduced the latency of sleeping time evoked by pentobarbital compared with the control group (P < 0.001). Doses of 100 or 1000 mg kg(-1) also increased the sleeping time induced by pentobarbital (30.9 +/- 3.91 and 52.1 +/- 15.6 min, respectively) compared with the negative control (12.4 +/- 4.27 min). We investigated the antinociceptive effect of the essential oil in chemical (acetic acid) and thermal (hot-plate) models of nociception in mice. Dipyrone (200 mg kg(-1)) and the highest doses of the essential oil (1000 mg kg(-1)) significantly inhibited acetic acid-induced abdominal constriction in mice (5.00 +/- 1.38 and 6.8 +/- 2.1 constrictions, respectively) compared with the negative control (33.1 +/- 2). The same dose of essential oil also raised the pain thresholds of mice in the hot-plate test and significantly (P < 0.05) increased the latency at all observation times. In acetic acid-induced abdominal constriction in mice pretreatment of the animals with naloxone (5 mg kg(-1)) significantly reversed the analgesic effect of morphine and of the essential oil at the highest dose (1000 mg kg(-1)). The essential oil of C. cajucara was also investigated for its anti-inflammatory properties. At the lowest dose (100 mg kg( 1)) the essential oil had anti-inflammatory effects in animal models of acute (carrageenin-induced paw oedema in mice) and chronic (cotton pellet granuloma) inflammation. The essential oil at doses of 50, 100 and 200 mg kg(-1) significantly and dose-dependently inhibited carrageenan-induced oedema (49 +/- 5; 37 +/- 5; 34 +/- 8 mg, respectively) compared with the negative control (74 +/ 8 mg). The essential oil (100 mg kg(-1)) also inhibited chronic inflammation by 38% whereas diclofenac inhibited it by 36%. However, the essential oil did not inhibit the migration of neutrophils into the peritoneal cavity. These data show that the essential oil from C. cajucara contains compounds that had a significant antinociceptive effect when the oil was administered at the highest dose. This effect seems to be related to interaction with the opioid system. The essential oil also had a significant anti-inflammatory effect in acute and chronic inflammation models when administered at lower doses. This effect seems to be related to cyclooxygenase inhibition. PMID- 10678503 TI - Investigation of plants used in Jamaican folk medicine for anti-bacterial activity. AB - We have started a systematic scientific study of folklore medicinal plants currently used as alternative medicine in Jamaican society. In this initial study, extracts of plants widely used by the islanders are studied for antibacterial activity against five common pathogens; Streptococcus group A, Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa and Escherichia coli. These studies revealed that 25% (approx.) of the plant extracts had antimicrobial activity against at least one of the microbes used. Subsequent to these observations, extracts from Mikania micrantha were examined in detail. This led to the isolation of two sesquiterpenoids, mikanolide and dihydromikanolide, with activity against S. aureus and C. albicans. The results suggest that traditional folk medicine could be used as a guide in our continuing search for new natural products with potential medicinal properties. PMID- 10678504 TI - Fluorimetric determination of ampicillin by use of non-toxic catalysts. Estimation of beta-lactamase activity and parameters. AB - A fluorescence assay based on the use of biological reducing agents as catalysts rather than heavy metal ions has been developed for estimation of ampicillin concentrations. The assay is based on the conversion of ampicillin to its penicilloate, by treatment with sodium hydroxide, then neutralization, dilution with 0.5 M acetate buffer at pH4 and heating for 30 min at 100 degrees C in the presence of ascorbic acid (0.5 mg) and EDTA (50 microM). Reduced glutathione, cysteine and N-acetylcysteine also catalysed the development of fluorescence. A practical sensitivity range of 0.5-50 microM ampicillin was used. The assay was used to estimate ampicillin in some biological solutions to which the antibiotic has been added. Milk, blood serum, trypticase soy broth and nutrient broth containing 25 microM antibiotic assayed at 18.5, 21.7, 16.5 and 14.7 microM, respectively, with standard deviations between 1.2 and 0.7%. The low results were attributed to binding of some ampicillin by proteins or peptides which were removed by pretreatment. Urine containing 5 mM ampicillin assayed at 4.97 mM with a standard deviation of 3%. A modification of the procedure was used to measure beta-lactamase activity against ampicillin in several organisms in a fixed time assay. Kinetic parameters of a commercial beta-lactamase preparation from Bacillus cereus could also be determined by an additional modification. In both instances a correction was required for the intrinsic fluorescence of ampicillin remaining in the solution. The preparation examined had a Michaelis constant (Km) of 0.32 mM, a maximum velocity (Vmax) of 5398 micromol ampicillin hydrolysed mg( 1) min(-1), an apparent catalytic constant (Kcat, turnover number) of 20,220 s( 1) and a Kcat/Km ratio of 0.63 x 10(7) M(-1) s(-1). The major advantage of using this assay technique is that toxic metals are not used in the development of fluorescence so it is more environmentally acceptable. The technique is useful for examining beta-lactamase activity against ampicillin and might be useful for pharmaceutical products-for determining available therapeutic levels and for monitoring the activity of penicillin acylase against the penicilloate of ampicillin. PMID- 10678505 TI - Application of magnetoencephalography to the study of autism. PMID- 10678506 TI - Cognitive correlates of the negative, disorganized, and psychotic symptom dimensions of schizophrenia. AB - Knowledge of the relationship between specific cognitive abnormalities and the clinical symptoms of schizophrenia could give insight into the nature of their underlying pathophysiology. Composite scores were generated for negative, disorganized, and psychotic symptom ratings in 134 patients with schizophrenia (DSM-IV criteria). Partial correlations (each composite corrected for the others) were computed with neuropsychological measures. Negative symptoms were related to poor performance on tests of verbal learning and memory, verbal fluency, visual memory, and visual-motor sequencing. Disorganized symptoms were correlated with lower verbal IQ and poor concept attainment. Psychotic symptoms had no significant relationship with cognitive deficit. PMID- 10678507 TI - Prevalence of obsessive-compulsive disorder in schizophrenia and significance of motor symptoms. AB - To investigate the differences between schizophrenic subjects with and without obsessive-compulsive disorder (OCD), the authors systematically assessed 76 schizophrenic subjects for OCD. Subjects with and without OCD were then compared with regard to motor symptoms, including catatonia, and several measures of psychopathology. Treatment strategies were evaluated retrospectively. The 12 subjects with OCD (15.8%) had more motor symptoms, including catatonia, than non OCD schizophrenic subjects. Some differences were found with regard to psychopathological symptoms. Treatment strategies also differed in the two groups. The high prevalence of motor symptoms in these subjects supports the hypothesis of a basal ganglia-frontal lobe connection linking OCD with schizophrenia. PMID- 10678508 TI - Occipital atrophy is associated with visual hallucinations in Alzheimer's disease. AB - In this study of patients with Alzheimer's disease (AD), patients with visual hallucinations were compared with patients who did not have visual hallucinations to determine if selective occipital lobe atrophy is associated with visual hallucinations. Seven AD patients with visual hallucinations were matched by cognitive score to 7 AD patients without visual hallucinations and 3-D MRI images obtained. A ratio of measured occipital volumes to whole brain volumes was compared between the two groups. AD patients with visual hallucinations had a significantly smaller occipital/whole brain ratio than AD patients without visual hallucinations. These results suggest visual hallucinations in AD may be associated with neuropathology of the occipital lobe. PMID- 10678509 TI - Olfactory dysfunction discriminates probable Alzheimer's dementia from major depression: a cross-validation and extension. AB - The present study was conducted to cross-validate and extend the hypothesis that olfactory dysfunction could discriminate between groups of patients with Alzheimer's disease and major depression. Forty patients meeting the DSM-IV criteria for either Alzheimer's disease or major depression (20 per group) underwent assessment with the Pocket Smell Test (PST), a three-item screening measure of odor identification, and the Mini-Mental State Examination (MMSE). A PST score of < or = 1 (1 or 0 correct) discriminated between the groups with a hit rate of 97.5% (sensitivity = 95%, specificity = 100%). The optimal hit rate for the MMSE (< or =24) was less effective (hit rate = 90%, sensitivity = 80%, specificity = 100%). Age, gender, and education had minimal impact on the PST for both groups. Olfactory assessment continues to add to the diagnostic utility in the differential diagnosis of Alzheimer's disease versus major depression in elderly patients. PMID- 10678510 TI - An instrumental study of the relationship between extrapyramidal signs and psychosis in Alzheimer's disease. AB - Neurobehavioral disturbances often coexist with extrapyramidal signs (EPS) in patients with Alzheimer's disease (AD). In the present study, we examined the relationships between delusions and hallucinations and EPS by using standard observer ratings and sensitive electromyographic (EMG) measures in 52 patients with probable AD. On the basis of observer ratings, 36.5% of the patients exhibited psychotic features and 63.5% exhibited parkinsonism. Severity of clinically rated parkinsonism and the EMG measure of bradykinesia were significantly correlated with severity of neurobehavioral disturbances in this sample. The association between parkinsonism and delusions and hallucinations suggests a subcortical mechanism in the etiopathology of psychosis in AD. PMID- 10678511 TI - Treatment of dementia-associated agitation with gabapentin. AB - The authors describe the use of gabapentin in the treatment of 4 outpatients with dementia-associated agitation. On the basis of clinical case reports and the Overt Agitation Severity Scale, all 4 patients had reduced agitation with gabapentin. Three of 4 patients were successfully titrated to a full dose of 2,400mg/day. These findings suggest a possible role for gabapentin in the behavioral management of patients with dementia. PMID- 10678512 TI - Normal cognitive performance in patients with chronic alcoholism in contrast to patients with Korsakoff's syndrome. AB - This study investigated which cognitive deficits are associated with chronic alcoholism. Neuropsychological profiles and MRI brain structure volumes of 14 patients with Korsakoff's syndrome, 15 patients with chronic alcoholism, and 16 healthy control subjects were compared. The patients with alcoholism had a normal cognitive performance and normal brain structure volumes. The patients with Korsakoff's syndrome had performance deficits on tests of memory, visuoperceptual, and executive functions, as well as reduced brain structure volumes. The results suggest that the cognitive deficits cannot be ascribed to mere chronic consumption of alcohol. If cognitive deficits are present in patients with chronic alcoholism, this may point to an underlying brain disorder. PMID- 10678513 TI - Relationship between symptoms and motoric subtype of delirium. AB - For 46 patients with delirium who were consecutive referrals to a consultation liaison psychiatry service, the authors describe the relationships between symptoms, as rated on the Delirium Rating Scale, and delirium motoric subtypes, as defined by Liptzin and Levkoff's criteria. Most cases were of the mixed subtype (46%), 24% were hypoactive, and 30% were hyperactive. Overall scores differed significantly among motoric subtype groups, being highest in the hyperactive, lowest in the hypoactive, and intermediate in the mixed. On item scores, the hypoactive group scored lower than the hyperactive group for delusions, mood lability, sleep-wake cycle disturbances, and variability of symptoms, but lower than the mixed group only for mood lability. The results suggest that delirium presents as motoric subtypes that differ according to symptom profile and severity of delirium. These subtypes may differ in their underlying pathophysiologies, responsiveness to therapeutic interventions, and outcome. PMID- 10678514 TI - Is delirium after cardiac surgery related to plasma amino acids and physical condition? AB - The authors studied interrelationships between plasma levels of amino acids, physical condition (as apparent from cortisol, albumin, and thyroid hormone concentrations), and postoperative delirium in 296 patients undergoing elective cardiac surgery. Both plasma tryptophan (Trp) and ratio of Trp to the other large neutral amino acids (oLNAA) were reduced in delirious patients compared with control patients. The lower availability of Trp for the brain in delirious patients may lead to decreased serotonergic function. Besides, the ratio of phenylalanine (Phe) to the oLNAA was increased in delirium, which may result in a higher synthesis of cerebral dopamine and norepinephrine. Delirious patients were also in poorer physical condition than nondelirious patients, having decreased albumin level and increased ratio of inactive reverse triiodothyronine (T3) to active T3. Decreased Trp and increased Phe availability may give rise to an imbalance in cerebral neurotransmitters and thus contribute to delirium. PMID- 10678515 TI - A neuropsychological comparison of depressed suicide attempters and nonattempters. AB - The neuropsychological performance of 18 older inpatients with major depression who were admitted following a suicide attempt was compared with that of 29 older depressed inpatients who had never attempted suicide. There was an interactive effect of age and group on the Trail Making Test, part B, such that attempters showed greater performance declines with age. No other differences were detected between groups on a range of neuropsychological tasks. These findings are discussed in the context of the methodological limitations of previous studies and the need for future research to better elucidate the nature of the relationships between age, cognitive functioning, and suicidal behavior. PMID- 10678516 TI - Pedophilia and temporal lobe disturbances. AB - Paraphilias may occur with brain disease, but the nature of this relationship is unclear. The authors report 2 patients with late-life homosexual pedophilia. The first met criteria for frontotemporal dementia; the second had bilateral hippocampal sclerosis. Both were professional men with recent increases in sexual behavior. In both, 18-fluorodeoxyglucose positron emission tomography revealed prominent right temporal lobe hypometabolism. These cases and the literature suggest that bilateral anterior temporal disease affecting right more than left temporal lobe can increase sexual interest. A predisposition to pedophilia may be unmasked by hypersexuality from brain disease. These observations have potential implications for all neurologically based paraphilias. PMID- 10678517 TI - Impaired auditory gating and P50 nonsuppression following traumatic brain injury. AB - Traumatic brain injury (TBI) can produce persistent attention and memory impairment that may in part be produced by impaired auditory sensory gating. The P50 evoked waveform response to paired auditory stimuli appears to be a useful measure of auditory gating. The first controlled measurement of the P50 ratio in TBI patients is described: when 20 patients with persistently symptomatic TBI were compared with 20 control subjects, the P50 ratio was significantly greater in the TBI group. The potential neurophysiologic and therapeutic implications of this finding in TBI patients who report symptoms consistent with impaired auditory gating are discussed. PMID- 10678518 TI - Interrelationships between nocturnal sleep, daytime alertness, and sleepiness: two types of alertness proposed. AB - The authors studied daytime sleepiness and alertness (based on the Multiple Sleep Latency Test [MSLT] and Maintenance of Wakefulness Test [MWT]) and nocturnal sleep in 22 patients with depression/anxiety and in 47 nondepressed patients with sleep apnea. The patients underwent two overnight sleep studies followed by daytime tests. In depressed patients, MWT scores correlated negatively with total sleep time and stage 3. MSLT scores correlated negatively with total sleep time and with sleep efficiency. Apneic patients showed a negative correlation between MWT results and amount of stage 1 sleep. MSLT results correlated positively with sleep onset latency on the preceding overnight sleep study. Thus, in depressed patients, there is a paradox that with more disturbed sleep there is greater daytime alertness. In contrast, the more disturbed the sleep is in sleep apnea patients, the more difficult it is to maintain daytime alertness. PMID- 10678519 TI - Clinical and neuropsychological profiles of obsessive-compulsive schizophrenia: a pilot study. AB - This pilot study compared characteristics of obsessive-compulsive (OC) schizophrenic patients and a matched non-OC schizophrenic control group. The OC schizophrenic group required more intensive clinical interventions and had a poorer clinical course, lower levels of functioning, and longer periods of hospitalization. They showed greater negative symptoms and more impaired executive functioning. These findings suggest OC-schizophrenic patients may have an atypical set of clinical and neuropsychiatric characteristics, perhaps constituting a subgroup within the schizophrenia spectrum. Pathophysiology and possible treatment implications require further study. PMID- 10678521 TI - Perceptual aberration and schizotypy: a cautionary note. AB - This study assessed 51 college students for associations between Continuous Performance Test performance and schizotypy scale scores. Results suggest that perceptual aberration scores, while generally correlated with overall schizotypy scores, may not be adequate as a single-criterion measure of schizotypy. PMID- 10678520 TI - Heritability heightens brain metabolite differences in schizophrenia. AB - Single-voxel proton magnetic resonance spectroscopy was performed in 64 medicated schizophrenic patients and 51 healthy subjects. Spectra were obtained from a voxel in the left medial temporal lobe by using a 2.0-tesla whole-body magnetic resonance imaging system. Schizophrenic patients showed a lower N acetylaspartate/ creatine-phosphocreatine ratio than did healthy subjects, and this reduction was greater in 13 patients with a family history of psychotic disorders. PMID- 10678522 TI - Treatment of pathological affect: variability of response for laughter and crying. AB - Pathological laughing and crying (PLC) is increasingly recognized to accompany diverse neurologic conditions, although it remains poorly understood. The authors describe 3 cases of amyotrophic lateral sclerosis (ALS) with an unusual change from a predominance of pathological crying to laughter following drug treatment. Possible explanations for this phenomenon are discussed. PMID- 10678524 TI - "Hyperacusis" and origins of lowered sound tolerance. PMID- 10678523 TI - Loss of recent memory after bilateral hippocampal lesions. 1957. AB - Bilateral medial temporal lobe resection in man results in a persistent impairment of recent memory whenever the removal is carried far enough posteriorly to damage portions of the anterior hippocampus and hippocampal gyrus. This conclusion is based on formal psychological testing of nine cases (eight psychotic and one epileptic) carried out from one and one-half to four years after operation. The degree of memory loss appears to depend on the extent of hippocampal removal. In two cases in which bilateral resection was carried to a distance of 8 cm posterior to the temporal tips the loss was particularly severe. Removal of only the uncus and amygdala bilaterally does not appear to cause memory impairment. A case of unilateral inferior temporal lobectomy with radical posterior extension to include the major portion of the hippocampus and hippocampal gyrus showed no lasting memory loss. This is consistent with Milner and Penfield's negative findings in a long series of unilateral removals for temporal lobe epilepsy. The memory loss in these cases of medial temporal lobe excision involved both anterograde and some retrograde amnesia, but left early memories and technical skills intact. There was no deterioration in personality or general intelligence, and no complex perceptual disturbance such as is seen after a more complete bilateral temporal lobectomy. It is concluded that the anterior hippocampus and hippocampal gyrus, either separately or together, are critically concerned in the retention of current experience. It is not known whether the amygdala plays any part in this mechanismi, since the hippocampal complex has not been removed alone, but always together with uncus and amygdala. PMID- 10678525 TI - Cotard's syndrome in a young male bipolar patient. PMID- 10678526 TI - The distributions of apoptotic cells in the medial preoptic areas of male and female neonatal rats. AB - The medial preoptic area (MPA) of the hypothalamus of the rat contains two sexually dimorphic nuclei, the periventricular preoptic nucleus (PVpo) and the medial preoptic nucleus (MPN). To examine the relationship between sexual dimorphism and neuronal death, we examined the number of apoptotic cells in the subdivisions of the MPA in neonatal rats of postnatal days 1 (P1), 4 (P4), 7 (P7) and 14 (P14). Apoptotic cells in these areas were classified according to their progression into three stages. P1 and P4 rats contained many apoptotic cells in the subfield along the third ventricle, including the PVpo, and their number was significantly larger in P1 males: in particular, the number of early-stage cells was larger in males than females. The number of apoptotic cells in the MPN was increased in P4 and P7 rats, although no significant sexual differences were seen in the total number or in the number of each progressive stage of apoptotic cells. In P14 rats, very few apoptotic cells were seen in the MPA. Our data revealed that the distribution of apoptotic cells in the MPA of developing rats depends on the sexuality, subdivision of the area and postnatal period. PMID- 10678527 TI - Hemispheric asymmetry and corpus callosum morphometry: a magnetic resonance imaging study. AB - Previous post-mortem studies (Aboitiz, F., Scheibel, A.B., Fisher, R.S., Zaidel, E., 1992. Brain Res. 598, 154-161 and Aboitiz, F., Scheibel, A.B., Zaidel, E., 1992. Brain 115, 1521-1541) have shown an inverse association between asymmetry in perisylvian areas and the size of a specific segment, the isthmus, of the corpus callosum (CC) in males. The purpose of this work was to study in vivo the association between hemispheric asymmetry and the total size of the CC in 35 right-handed subjects (16 males, 19 females; mean age 24.9 +/- 3.9). An MRI scan was performed for each subject. The area of the right (RH) and left (LH) hemispheres were measured from images in the sagittal plane and the area of the CC from images in the mid-sagittal plane. The index of hemispheric asymmetry was absolute value((LH - RH)/[(LH + RH)/2]). There was a negative correlation between the absolute value of hemispheric asymmetry and the size of the CC in males (r = 0.55, P = 0.03) but not in females (r = -0.20, P = 0.42). These findings, like those of Aboitiz et al. (Aboitiz, F., Scheibel, A.B., Zaidel, E., 1992. Brain 115, 1521-1541), suggest a sex-dependent decrease in interhemispheric connectivity with increasing hemispheric asymmetry. PMID- 10678528 TI - Direct projection from the cardiovascular control region of the cerebellar cortex, the lateral nodulus-uvula, to the brainstem in rabbits. AB - In decerebrate unanesthetized rabbits, electrical stimulation of the lateral nodulus-uvula in the cerebellar vermal cortex evoked an increase in renal sympathetic nerve activity, an increase in blood pressure and a decrease in renal arterial blood flow, which were all in contrast to the effects reported previously in the anesthetized rabbits. In order to identify the pathway mediating these responses, we investigated the Purkinje cell projection from the lateral nodulus-uvula using both anterograde (biotinylated dextran amine, BDA) and retrograde (horseradish peroxidase, HRP) tracing methods in rabbits. When BDA was iontophoretically injected into the lateral nodulus-uvula, labeled Purkinje cell axons were found within and around the superior and inferior cerebellar peduncles (SCP and ICP, respectively). Furthermore, terminal-like fields were found in the dentate and vestibular nuclei as reported in previous studies. However, the terminal-like patterns that we observed in the parabrachial nucleus (PB) in the rabbit have not been reported yet. When HRP was microinjected into the lateral PB, retrogradely labeled Purkinje cells were found in the lateral nodulus-uvula. These results indicate that Purkinje cells in the lateral nodulus uvula project into the vestibular nuclei via the ICP and to the lateral PB via the SCP. We suggest that these two pathways mediating cardiovascular responses have different sensitivities to anesthetics. PMID- 10678529 TI - Suppression on neuronal responses by a metacontrast masking stimulus in monkey V4. AB - We studied the temporal characteristics of suppression in area V4 of the monkey using a visual stimulus for metacontrast masking. Visual responses of V4 neurons to a brief test stimulus presented within the receptive field were recorded, and the effect of a mask stimulus that did not spatially overlap the test stimulus was examined. Responses to the test stimulus were suppressed by the mask stimulus, which either preceded or followed the test stimulus. To study the temporal characteristics of suppression, the interval between the onset of the test stimulus and that of the mask stimulus (stimulus onset asynchrony, SOA) was varied. Maximum suppression occurred with a simultaneous presentation of the two stimuli, and the suppression gradually weakened as the SOA increased. The suppressive effect of the mask stimulus lasted on average about 77 ms in the negative SOA (forward masking) and 65 ms in the positive SOA (backward masking). These results indicate that surround suppression in V4 neurons has considerable temporal width, which is longer than that previously reported in areas V1 and V2. There were marked differences between the time course of suppression in V4 neurons in the present study and those reported in human metacontrast masking. PMID- 10678530 TI - Interaction between nitric oxide and substance P on heat-induced inflammation in rat paw. AB - To elucidate the interaction between nitric oxide (NO) and substance P (SP) in neurogenic inflammatory responses, we measured the change in the degree of Evans blue leakage and NO levels in perfusate from the subcutaneous space in the rat instep following noxious heat stimulation (47 degrees C for 30 min). Furthermore, the effects of drugs affecting nitric oxide synthase were examined. Noxious heat stimulation caused on an increase in NOx, or NO2- and NO3- into the perfusate in parallel with plasma extravasation. Nw-nitro-L-arginine methylester (L-NAME: 100 mg/kg once daily.) intraperitoneally (i.p.) given five times (chronic treatment) significantly suppressed the increase in Evans blue extravasation induced by heat stimulation, whereas acute treatments with L- and D-NAME (100 mg/kg once, i.p.) did not show any significant effect. NO release induced by heating also was significantly suppressed by chronic pretreatment with L-NAME, but not by acute treatment. SP (10(-5) M) applied into the perfusate caused a remarkable increase in the NOx release into the perfusate. Intra-arterial injection of RP67580 (1 mg/kg) on the perfused side, but not SR48968 (1 mg/kg), significantly attenuated the increases in Evans blue leakage and NOx release during heat stimulation. These results suggest that heat-induced SP release from the peripheral endings of small-diameter afferent fibers causes NO generation through NK-1R, and that this gas act to elicit or enhance inflammatory responses. PMID- 10678531 TI - Intrinsic and commissural connections within the entorhinal cortex. An anterograde and retrograde tract-tracing study in the cat. AB - Intrinsic and commissural connections within the entorhinal cortex (EC) were examined in the cat by the anterograde and retrograde tract-tracing methods with Phaseolus vulgaris leucoagglutinin and cholera toxin B subunit. Intrinsic axons to the superficial layers (layers I-III) arose mainly from layers II, III, Vd (deep part of layer V), and VI, were distributed more widely in the superficial layers than in the deep layers, and terminated progressively more densely in more superficial layers; most densely in layer I. In the medial entorhinal area (MEA) and the ventromedial and the ventrolateral divisions of the lateral entorhinal area (VMEA and VLEA), the longitudinal connections through the intrinsic fibers to the superficial layers is often more restricted in rostral direction than in caudal direction. In the dorsolateral division of the lateral EC (DLEA), the longitudinal connections through the intrinsic fibers to the superficial layers extended distantly in both rostral and caudal directions. Intrinsic fibers to the deep layers (layers IV-VI) originated mainly from layers IV and Vs (superficial part of layer V) and were distributed rather sparsely and diffusely; they were distributed more widely in the deep layers than in the superficial layers. Commissural axons to the homotopic EC regions originated from layers II and III of the MEA and DLEA and terminated in all EC layers, most densely in layer I. PMID- 10678532 TI - Histamine excites rat cerebellar Purkinje cells via H2 receptors in vitro. AB - Recent neuroanatomical studies have revealed a direct hypothalamocerebellar histaminergic pathway. However, the functional significance of the histaminergic fibers in the cerebellum is not yet clear. In this study, the effects of histamine on the firing of cerebellar Purkinje cells (PCs) were investigated in vitro. Histamine predominantly produced excitatory (106/111, 95.5%) and in a few cases inhibitory (5/111, 4.5%) responses in PCs. The histamine-induced excitation was not blocked by perfusing the slice with low Ca2+ high/Mg2+ medium (n = 8), supporting a direct postsynaptic action of histamine. The histamine H2 receptor antagonist ranitidine effectively blocked the excitatory response of PCs to histamine (n = 20), but triprolidine, an H1 receptor antagonist, could not significantly block the histamine-induced excitation, or only very slightly decreased the excitatory effect of histamine on the cells (n = 13). On the other hand, the highly selective H2 receptor agonist dimaprit mimicked the excitatory effect of histamine on PCs and this dimaprit-induced excitation was also blocked by ranitidine (n = 20), but not triprolidine (n = 8). However, the H1 receptor agonists betahistine and 2-thiazolylethylamine did not show any effect on the PCs (n = 9 and 14). These results reveal that histamine excites cerebellar PCs via H2 receptors and suggest that the hypothalamocerebellar histaminergic fibers may play an important role in functional activities of the cerebellum. PMID- 10678533 TI - Heterosynaptic expression of depolarization-induced suppression of inhibition (DSI) in rat hippocampal cultures. AB - Depolarization-induced suppression of inhibition (DSI) is a transient suppression of the inhibitory synaptic transmission, observed in the hippocampus and the cerebellum, upon postsynaptic depolarization. Using rat hippocampal cultures, we examined whether DSI is confined to the inhibitory synapses on the depolarized neuron or, if DSI can spread to those on neighboring non-depolarized neurons. Whole-cell recordings were performed in 108 neuronal pairs with the following synaptic responses. Stimulation of one neuron evoked the inhibitory autaptic currents (IACs) recurrently in that neuron and also elicited the inhibitory postsynaptic currents (IPSCs) orthodromically in the other neuron. In 38 of 108 pairs, the postsynaptic depolarization caused transient suppression of IPSCs (homosynaptic DSI). In 11 of the 38 pairs exhibiting the homosynaptic DSI, the depolarization also induced suppression of IACs (heterosynaptic DSI). The heterosynaptic DSI, like the homosynaptic DSI, depended on depolarizing pulse duration and was blocked by a phorbol ester. These results suggest that DSI can spread to the synapses on a neighboring non-depolarized neuron in rat hippocampal cultures. PMID- 10678534 TI - Directional tuning profiles of motor cortical cells. AB - The directional tuning profiles of motor cortical cells are commonly described by a cosine tuning function with three adjustable parameters (Georgopoulos, A.P., Kalaska. J.F., Crutcher, M.D., Caminiti, R., Massey, J.T., 1982. On the relations between the direction of two-dimensional (2D) arm movements and cell discharge in primate motor cortex. J. Neurosci. 2, 1527-1537). In this study the variation in the shape of the directional tuning profiles among a population of cells recorded from the arm area of the motor cortex of monkeys using movements in 20 directions, every 18 degrees, was examined systematically. This allowed the investigation of tuning functions with extra parameters to capture additional features of the tuning curve (i.e. tuning breadth, symmetry, and modality) and determine an 'optimal' tuning function. These functions provided better fit than the standard cosine one. The optimal function for the large majority of tuned cells was unimodal (84%), and only for a few of them (16%) it was bimodal. Of the unimodal cells, 73% exhibited symmetric and 27% asymmetric shape. The half-width, sigma, at the midpoint of optimal tuning curves differed among cells from 30 to 90 degrees, with a median at 56 degrees. This is much narrower than in the standard cosine tuning function with a fixed width of sigma = 90 degrees. It was concluded that motor cortical cells are more sharply tuned than previously thought. PMID- 10678535 TI - When pyramidal neurons lock, when they respond chaotically, and when they like to synchronize. AB - We give an overview on the locking properties of perturbed regularly firing pyramidal neurons, as a function of perturbation strength, self-spiking frequency and perturbation frequency. For inhibitory perturbations, instead of locking chaotic response emerges for a whole range of parameters. This suggests that global synchronization on the set of inhibitory connections may easily be achieved. PMID- 10678536 TI - A new technique for implanting a fine-wire microelectrode for chronic recording of unit activity from freely-moving mice. AB - We report here a newly developed chronic implantation technique using an epoxy coated fine-stainless steel wire (33 microm in diameter) to record single unit activity from the brain of freely-moving mice with as little tissue injury as possible. Since the fine-wire electrode is not capable of staying straight by itself or of penetrating into the brain, a pair of permanent neodymium magnets placed on a micromanipulator as well as below the animal's head was used for stereotaxic implantation to keep the fine-wire straight and strong by the magnetic fields. With those implanted electrodes recording of single units from the hippocampal CA1 of freely-moving mice was performed during sleep and wakefulness. PMID- 10678537 TI - Should we still use nitrovasodilators to test baroreflex sensitivity? PMID- 10678538 TI - How to measure baroreflex sensitivity: from the cardiovascular laboratory to daily life. AB - Arterial baroreflex function in humans is commonly assessed through a number of laboratory tests based on quantification of the reflex responses in heart rate or blood pressure to external stimuli applied to the cardiovascular system. Evidence is available that these laboratory estimates of baroreflex sensitivity have both pathophysiological and clinical relevance. Indeed, a number of studies have shown that the sensitivity of the baroreceptor-heart rate reflex may have a prognostic value in myocardial infarction, heart failure and diabetic patients, where mortality seems to be inversely related to the sensitivity of cardiac baroreflex modulation. A deeper insight into the features of daily-life baroreflex cardiovascular control has been offered more recently by techniques based on computer analysis of spontaneous blood pressure and heart rate fluctuations. This innovative approach allows spontaneous baroreflex sensitivity to be assessed in real life conditions, with no need for external stimulation of the patient as required by the older laboratory techniques. This review will briefly survey the methods most widely used to assess baroreflex function in humans, in the laboratory and in daily life. PMID- 10678539 TI - Controlled study of circadian rhythm of blood pressure in patients with aldosterone-producing adenoma compared with those with essential hypertension. AB - OBJECTIVE: The circadian rhythm of blood pressure in patients with aldosterone producing adenoma (APA) is not well understood. We evaluated the circadian blood pressure rhythm in such patients by comparison with that in patients with essential hypertension (EHT). The latter are characterized by a nocturnal blood pressure decline, the so called 'dipping' blood pressure pattern. DESIGN AND METHODS: A total of 12 patients with APA and 36 patients with EHT who were matched by age and sex, and who had no severe organic disorders, were hospitalized to control their diet (low sodium) and activities. Ambulatory blood pressure monitoring was conducted for 24 h. The 24-h blood pressure was divided into waking blood pressure (0600-2130 h) and sleeping blood pressure (2200-0530 h). RESULTS: The two groups showed no significant differences in age, sex, serum creatinine, plasma glucose, daily urinary sodium excretion, and left ventricular mass index. Although the 24-h mean blood pressure was higher in APA (112 +/- 8 mmHg) than EHT (102 +/- 12 mmHg), the dipping mean blood pressure ratio (%), which was calculated from the sleeping and waking blood pressures, did not differ significantly between the two groups (93.2 +/- 5.4 versus 92.8 +/- 5.9). CONCLUSION: The dipping ratio of blood pressure in patients with APA resembled that of patients with EHT. Variables that would influence the circadian rhythm of blood pressure were controlled during study. The results suggest that a circadian blood pressure in patients with APA is of the dipping type, characterized by a nocturnal blood pressure decline, when a low sodium diet is ingested. PMID- 10678540 TI - Blood pressure and menopausal transition: the Atherosclerosis Risk in Communities study (1987-95). AB - OBJECTIVE: Blood pressure changes during menopausal transition have not been studied previously using a biracial sample. We investigated whether menopausal transition was associated with change in blood pressure in African-American or white women. DESIGN, SETTING AND PARTICIPANTS: The prospective multicenter study, the Atherosclerosis Risk In Communities (ARIC) Study (1987-95) was utilized. Included were never-users of hormone replacement therapy (3,800 women, 44% of the original sample). MAIN OUTCOME MEASURE: Changes in blood pressure were adjusted for baseline age and body mass index, baseline blood pressure, antihypertensive use, ARIC field center and weight change. The menopausal transition group was compared to the non-transition group, separately, by ethnicity. RESULTS: Women undergoing the menopausal transition did not differ significantly in regard to systolic blood pressure change [5.2, 95% confidence interval (CI) 4.0-6.4] from non-transitional women (4.6, 95% CI 4.0-5.2); adjustment for age, baseline systolic blood pressure and other factors did not alter this finding. Transitional women had significantly less diastolic blood pressure change (-0.5, 95% CI -1.1 to 0.2) than non-transitional women (-2.0, 95% CI -2.4 to -1.7, P= 0.000) but, after adjustment for other covariates, the result was not significant African-American women had significantly (P= 0.003) higher systolic blood pressure change compared to white women, but this difference became non significant (P= 0.21) after restricting the sample to women younger than 55 years of age. Interactions between menopausal transition and ethnicity were not significant, either in systolic blood pressure or diastolic blood pressure change. CONCLUSION: Menopausal transition is not associated with significant blood pressure change in African-American or white women. PMID- 10678541 TI - Effect of bisoprolol and atenolol on endurance exercise capacity in healthy men. AB - OBJECTIVES: To compare the effects of a highly beta1-selective adrenoceptor antagonist bisoprolol with those of atenolol and placebo on endurance exercise capacity in young, healthy male volunteers. DESIGN: Twelve subjects randomly received oral placebo, atenolol (100 mg/day) or bisoprolol (10 mg/day) for 3 weeks, following a double-blind cross-over design. METHODS: At the end of each period, the subjects performed an endurance exercise test on the bicycle ergometer at 70% of maximal aerobic power. Cardiac output was measured by means of an automated CO2-rebreathing method. Venous blood was sampled before, during and after exercise. RESULTS: Exercise duration was not significantly different between the two drugs tested. Total exercise duration was significantly reduced by bisoprolol (-19.4 +/- 6.7%, P< 0.01) (mean +/- SEM) and by atenolol (-29.8 +/- 6.6%, P< 0.001), compared with placebo. Atenolol and bisoprolol were equally effective in lowering resting plasma renin activity, heart rate and systolic blood pressure. Resting and exercise stroke volume were significantly increased by both drugs, so that cardiac output was not significantly affected. Both drugs induced significant decreases in plasma-free fatty acid concentrations during recovery and blunted the exercise-induced increase. There were no significant relationships between the reduction of exercise duration and the haemodynamic changes or the degree of impairment of the exercise-induced increase in free fatty acid release resulting from beta-blockade. CONCLUSIONS: It is concluded that both drugs affect endurance exercise capacity in young, normotensive men, with a tendency to a smaller reduction during bisoprolol treatment. Haemodynamic variables are unlikely to be involved in the reduction of endurance exercise capacity. The role of the reduced availability of plasma free fatty acids remains unclear. PMID- 10678542 TI - Fetal versus maternal determinants of the reduced fetal and placental growth in spontaneously hypertensive rats. AB - OBJECTIVE: Epidemiological studies indicate that a reduced birth weight and enlarged placenta increase the likelihood of human cardiovascular disease later in life. The relative importance of fetal versus maternal factors in these phenomena is not known. To assess the relative role of genotypic versus environmental factors in this effect, we examined whether the altered fetal and placental growth rates and amniotic fluid volume of spontaneously hypertensive rat (SHR) fetuses of the Okamoto strain, are modified by gestation in normotensive Wistar- Kyoto (WKY) rat mothers and vice versa. DESIGN: One-day-old SHR embryos were gestated for 16 days in either SHR or WKY recipients. Similarly, 1-day-old WKY rat embryos were gestated for 16 days in either SHR or WKY surrogates. At 16 days, fetal and placental weights were recorded. Paternal and maternal donor and recipient blood pressures, maternal body weight and average litter size within the four groups were also studied. METHODS: One cell SHR and WKY embryos were harvested from timed matings and transferred to psuedopregnant mothers of the same or opposite strain. Timed matings required routine vaginal smears for the detection of proestrus and the presence of sperm following overnight matings. Harvested embryos were temporarily maintained in culture medium in a 37 degrees C incubator until injection into the oviduct of recipients. Blood pressures were meaured using indirect tail-cuff plethysmography and a computerized data acquisition system. RESULTS: SHR fetal and placental weights at 16 days gestation were significantly lower than WKY fetal and placental weights, irrespective of maternal strain. At 16 days of gestation, the fetal and placental weights of SHR fetuses gestated in WKY rat surrogate mothers (0.21 +/- 0.01 g and 0.19 +/- 0.01 g, respectively) were not significantly different from those of SHR gestated in a surrogate SHR mother (0.21 +/- 0.01 g and 0.18 +/- 0.01 g, respectively). Similarly, the fetal and placental weights of WKY fetuses gestated in a WKY rat (0.27 +/- 0.01 g and 0.25 +/- 0.01 g, respectively) were unaltered by gestation in a SHR recipient (0.25 +/- 0.01 g and 0.23 +/- 0.01 g, respectively). The amniotic fluid volumes of SHR gestated in WKY rats and those of WKY fetuses gestated in SHR were not significantly different to each other (0.37 +/- 0.01 ml versus 0.38 +/- 0.01 ml, respectively) and were intermediate between the values for SHR and WKY fetuses gestated in a mother of the same strain (0.34 +/- 0.01 ml versus 0.44 +/- 0.02 ml, respectively). CONCLUSION: The SHR fetus exhibited reduced growth rate and placental size irrespective of maternal surrogate strain, suggesting that these measures are likely to be determined by the fetus or the placenta and, presumably, are independent of maternal blood pressure or altered electrolyte and hormonal milieu. PMID- 10678543 TI - Effect of ACE inhibition and angiotensin AT1 receptor blockade on renal and blood pressure response to L-arginine in humans. AB - OBJECTIVE: Nitric oxide (NO) may contribute to the actions of angiotensin converting enzyme (ACE) inhibitors. In contrast, angiotensin type 1 (AT1) receptor blockers (AT1B) have been considered to act exclusively by inhibiting angiotensin II actions. However, recent experimental findings suggest that AT1B actions may be also partly mediated by NO. In this study, we explored whether ACE inhibitors and AT1B modulate hemodynamic responses to L-arginine (L-arg), a NO precursor. METHODS: Systemic (Finapres) and renal hemodynamic responses to L-arg (200 mg/kg body weight), associated with markers of systemic and renal NO production, were assessed before (control) and after 3 weeks of randomized pretreatment with the ACE inhibitor ramipril (5 mg/day for 3 weeks) or the AT1B losartan (50 mg/day for 3 weeks) in nine healthy male subjects (33 +/- 2 years; body mass index 25.5 +/- 0.5 kg/m2). RESULTS: Control L-arg did not influence mean arterial pressure (MAP) (92 +/- 5 versus 90 +/- 5 mmHg; not significant). In contrast, L-arg decreased MAP when administered after pretreatment with ramipril (89 +/- 5 versus 83 +/- 4 mmHg; P< 0.01) or losartan (90 +/- 44 versus 86 +/- 4; P< 0.05). Control L-arg infusion had no effect on renal plasma flow (RPF) (paraminohippuric acid clearance) and renal vascular resistance (RVR), whereas the glomerular filtration rate (GFR) (inulin clearance) decreased (98 +/- 4 versus 89 +/- 5 ml/min; P< 0.05), resulting in a decrease in filtration fraction (P< 0.05). After ramipril, L-arg induced renal vasodilation as indicated by significant changes in RPF (576 +/- 41 versus 669 +/- 21 ml/min; P< 0.01) and RVR (P< 0.05). The GFR did not change statistically after ramipril pretreatment (91 +/- 3 versus 97 +/- 4 ml/min; not significant); however, the trend was different as compared with control (F= 5.7, P < 0.05). L-Arg-induced renal vasodilation was also observed after losartan (RPF, 637 +/- 34 versus 706 +/- 40 ml/min; P< 0.05). Enhanced renal and systemic responses to L-arg after ACE inhibitor and AT1B were associated with a rise in plasma L-citrulline levels, which was greater than after control L-arg (P < 0.05). However, other indicators of NO activity such as plasma and urinary cyclic guanosine 3',5'-monophosphate, and nitrates, remained unchanged throughout all experiments. CONCLUSION: The results indicate that ACE inhibitors and AT1B have a potential to enhance L-arg-induced vasodilation both in systemic and renal vascular beds. However, these hemodynamic responses were not associated with convincing changes in indicators of systemic or renal NO activity, suggesting a contribution of NO-independent vasodilator mechanisms. PMID- 10678544 TI - Adrenomedullin selectively inhibits angiotensin II-induced aldosterone secretion in humans. AB - OBJECTIVE: Adrenomedullin inhibits angiotensin II stimulated aldosterone production in vitro and in vivo in experimental animals. The aim of this study was to investigate the effect of adrenomedullin on angiotensin II and adrenocorticotrophic hormone-stimulated aldosterone production in vivo in healthy humans. DESIGN AND METHODS: Seven volunteers were studied in a quiet, temperature controlled laboratory. After 35 min of rest, an infusion of placebo or adrenomedullin (3 pmol/kg per min) was given over 60 min; 15 min after starting this first infusion, a second infusion of angiotensin II (0.96 fmol/kg per min) or adrenocorticotrophic hormone (0.1 mIU/kg per min) was co-infused and continued for 45 min. RESULTS: Adrenomedullin significantly inhibited angiotensin II stimulated aldosterone production: the increment in aldosterone on the placebo day was 691 pmol/l compared with 552 pmol/l on the adrenomedullin day (P< 0.004). Adrenomedullin did not inhibit adrenocorticotrophic hormone-stimulated aldosterone or cortisol release. CONCLUSION: Adrenomedullin selectively inhibits angiotensin II-stimulated aldosterone production. PMID- 10678545 TI - Postural hypotension following N-type Ca2+ channel blockade is amplified in experimental hypertension. AB - OBJECTIVE: To determine the relative importance of the cardiac and vascular sympathetic components of the orthostatic response to 90 degrees head-up tilt after N-type calcium-channel blockade in normotensive (sham renal cellophane wrap) and hypertensive (renal wrap) conscious rabbits. METHODS: The effects of N type calcium-channel blockade with omega-conotoxin GVIA (omega-CTX, 10 microg/kg i.v. bolus) were assessed in the absence or presence of cardiac block by propranolol and methscopolamine. These were contrasted with the effects of alpha1 adrenoceptor antagonism (prazosin 0.5 mg/kg i.v. bolus, in the presence of cardiac block) or ganglion blockade (mecamylamine 4 mg/kg i.v. bolus). RESULTS: In vehicle (0.9% saline) treatment groups, the response to tilt consisted of a small pressor effect (4 +/- 2 and 7 +/- 1 mmHg) and tachycardia (29 +/- 6 and 17 +/- 6 beats/min) in sham (n = 6) and wrap (n = 5) rabbits, respectively. After prazosin administration (with cardiac block), there were significant falls in MAP of 3 +/- 1 and 7 +/- 2 mmHg in sham (n = 7) and wrap (n = 6) rabbits, respectively, in response to tilt omega-CTX caused postural hypotensive responses of 8 +/- 2 and 13 +/- 2 mmHg in sham (n = 6) and wrap (n = 7) rabbits, respectively, and 7 +/- 1 and 14 +/- 2 mmHg in sham (n = 7) and wrap (n = 7) rabbits with prior cardiac block. Similarly, mecamylamine caused falls in MAP of 8 +/- 1 and 10 +/- 2 mmHg in response to tilt in sham (n = 6) and wrap (n = 9) animals, respectively. CONCLUSION: Sympathetic vasoconstrictor effectors are primarily responsible for maintaining blood pressure during tilt in conscious rabbits. The postural hypotension caused by sympatholytic agents is about double in hypertensive rabbits, and N-type calcium-channel blockade is as effective as ganglion blockade at inducing this syndrome. PMID- 10678546 TI - Relative influence of insulin resistance versus blood pressure on vascular changes in longstanding hypertension. ICARUS, a LIFE sub study. Insulin Carotids US Scandinavia. AB - BACKGROUND: Insulin resistance is associated with hypertension. The relative influences of hyperinsulinaemia and high blood pressure on vascular hypertrophy and carotid distensibility is unclear in patients with longstanding hypertension. METHODS: In 88 unmedicated patients with stage II-III hypertension and left ventricular hypertrophy on electrocardiogram we measured blood pressure, minimal forearm vascular resistance (MFVR) using plethysmography, intima-media thickness (IMT) and the wall distensibility of the common carotid arteries using ultrasound, and insulin sensitivity using a 2-h isoglycaemic hyperinsulinaemic clamp. RESULTS: IMT was positively correlated to systolic blood pressure (r= 0.26, P < 0.05), whole body glucose uptake index (M/IG; r= 0.22, P< 0.05), age (r= 0.24, P< 0.05) and negatively correlated to body mass index (r= -0.24, P < 0.05); IMT did not correlate to fasting serum insulin (r= -0.14, NS). In men (n = 64) MFVR was positively correlated to systolic blood pressure (r = 0.30, P < 0.05), but was unrelated to M/G and serum insulin. The distensibility of the common carotid arteries was negatively correlated to systolic blood pressure (r = -0.40, P< 0.001) and in untreated patients (n = 22) positively correlated to M/IG (r = 0.47, P < 0.05). CONCLUSIONS: High systolic blood pressure was related to vascular hypertrophy, whereas hyperinsulinaemia and insulin resistance were not, suggesting that longstanding high blood pressure is a far more important determinant for structural vascular changes than insulin resistance at this stage of the hypertensive disease. However, hyperinsulinaemia and insulin resistance were associated with low distensibility of the common carotid arteries in the subgroup of never treated hypertensive patients. PMID- 10678547 TI - Insulin levels during fasting and the glucose tolerance test and Homa's index predict subsequent development of hypertension. AB - OBJECTIVE: To determine whether there is a longitudinal relationship between hypertension and hyperinsulinemia and to find the most useful parameter(s) for predicting the subsequent development of hypertension. SUBJECTS AND METHODS: The oral glucose (75 g) tolerance test (OGTT) was performed in 313 patients, who were divided into three groups according to glucose tolerance based on the WHO criteria: normal, borderline and diabetes mellitus. The fasting insulin (IRI) levels, sigmaIRI (the sum of the insulin levels 0, 30, 60 and 120 min after the OGTT), insulinogenic index and Homa's index, a candidate for the simple assessment of insulin sensitivity, of the normotensive and hypertensive subjects in each subgroup were compared. In addition, 145 normotensive subjects were followed up for over 3 years and observed for the development of hypertension. RESULTS: Hypertensive diabetic subjects had not only higher fasting IRI levels and sigmaIRI values, but they also had higher Homa's indices than normotensive diabetics. Normotensive subjects with normal glucose tolerance (n = 20) did not develop hypertension. However, 16 out of 94 patients with borderline glucose tolerance and five out of 31 diabetics became hypertensive. The incidence of hypertension in the group with fasting IRI > or = 15, sigmaIRI > or = 150 or Homa's index > or = 4 was between 5 and 9 times higher than that in the group with fasting IRI < 10, sigmaIRI < 100 or Homa's index < 2. This difference was still significant when multivariate analysis, including various factors such as age, body mass index (BMI) and sex, was performed. CONCLUSIONS: These results suggest that higher plasma IRI levels and/or insulin resistance are closely related to the pathogenesis of hypertension in patients with diabetes mellitus. Homa's index, fasting and sigmaIRI may be useful predictors of the subsequent development of hypertension. PMID- 10678548 TI - Safety of the combination of valsartan and benazepril in patients with chronic renal disease. European Group for the Investigation of Valsartan in Chronic Renal Disease. AB - OBJECTIVE: Several experimental and clinical studies indicate that the renin system may play a pivotal role in progressing renal disease. The combination of an angiotensin-converting enzyme inhibitor and an angiotensin receptor blocker could provide a higher degree of blockade of the renin-angiotensin system than either agent alone. Such enhanced suppression might be of benefit for patients exhibiting a progressive decline in renal function because of chronic renal disease. METHODS: A pilot multinational, multicentre, randomized, active controlled, parallel group open-label study has been conducted in a group of patients with progressive chronic renal failure (creatinine clearance 20-45 ml/min) either with or without proteinuria and hypertension. The primary aim of the study was to investigate the safety and tolerability of the combination of valsartan and benazepril. Patients were randomly assigned to one of three groups: group 1 received valsartan 160 mg once daily (n = 22); group 2 received valsartan 80 mg once daily plus benazepril 5 or 10 mg once daily (n = 42); group 3 received valsartan 160 mg once daily plus benazepril 5 or 10 mg once daily (n = 44). The study lasted for 5 weeks, and in groups 2 and 3 benazepril was added on top of valsartan after the first week of therapy with the angiotensin receptor blocker. RESULTS: Serum creatinine increased in all three groups (mean change within a group: 11 micromol/l in group 1, P= 0.045; 9 micromol/l in group 2, P= 0.030; 15 micromol/l in group 3, P= 0.0006). Serum potassium also increased in all three groups of patients (mean change within a group: 0.28 mmol/l in group 1, P= 0.28; 0.48 mmol/l in group 2, P= 0.0008; 0.36 mmol/l in group 3, P= 0.02). After 5 weeks of treatment, the largest decrease in blood pressure was observed in group 3 (the mean change from baseline in seated diastolic blood pressure (SDBP) and seated systolic blood pressure (SSBP), respectively, were: -2.0 and -11.5 mmHg in group 1; -7.6 and -15.4 mmHg in group 2; -12.6 and -21.6 mmHg in group 3). In addition, both combination treatments resulted in the reduction of proteinuria. The total number of patients with adverse experiences were 10 (45.5%), 14 (33.3%) and 11 (25%) in groups 1,2 and 3, respectively. In six patients (5.6%) therapy was discontinued as a result of adverse experiences. Only one patient in each of the combined therapy groups withdrew from the study because of hyperkalaemia and no patients were forced to withdraw because of an increase in serum creatinine, acute renal failure or hospitalization. CONCLUSIONS: These results indicate that short-term combination of an angiotensin-converting enzyme inhibitor and an angiotensin receptor blocker is safe and well tolerated in patients with moderate chronic renal failure. PMID- 10678549 TI - Serum urate is associated with baseline renal dysfunction but not survival or deterioration in renal function in malignant phase hypertension. AB - BACKGROUND: There has been speculation whether serum uric acid levels are an independent prognostic factor in patients with hypertension. OBJECTIVE: To investigate the clinical associations and prognostic value of serum urate in patients with malignant phase hypertension (MHT), by comparing clinical features in patients with serum urate levels above and below the median levels for this population, and secondly, by performing a survival analysis in these patients. PATIENTS AND METHODS: Review of the data on 153 patients (98 males; mean age 50.3 years, SD 13.5) with MHT on the west Birmingham MHT register. Median uric acid levels in this population was 0.41 mmol/l (6.9 mg/dl), with an interquartile range of 0.34-0.50 mmol/l (5.7-8.4 mg/dl). Clinical characteristics of patients with a serum urate <0.41mmol/1 (group 1) were compared to those with levels above the median (0.41 mmol/l, group 2). RESULTS: Mean duration of follow-up was similar in both groups. The mean diastolic blood pressure at presentation and both mean systolic and diastolic blood pressures at follow-up were significantly higher in group 2 (that is, those with high serum urate levels) (unpaired t test, P= 0.039). There was also more renal dysfunction in group 2 patients with MHT, with higher mean serum urea and creatinine levels, both at presentation and at follow-up (unpaired t test, P< 0.01). The commonest causes of death were myocardial infarction (n = 7), heart failure (n = 4), stroke (n = 10) and renal failure (n = 5). There was no difference in mean survival duration between groups 1 and 2 (Kaplan-Meier, 64.6 versus 66.8 months; log-rank test, P= 0.519). Serum urate levels also did not predict the rise in serum creatinine levels (log-rank test, P= 0.84) or urea (P= 0.4033) amongst these patients. Using a multivariate Cox proportional hazards analysis, the only independent predictors of outcomes (death or the need for dialysis) were age (P = 0.007) and serum creatinine levels at presentation (P = 0.0046). CONCLUSION: Our analysis of a large series of patients with MHT shows that those with high urate levels had higher diastolic blood pressures and greater renal impairment at baseline. At follow-up, patients with median serum urate >0.41 mmol/l showed a greater deterioration in renal function and higher blood pressures, but no significant difference in survival. Serum urate levels also do not appear to be predictive of the deterioration in renal function or overall survival in patients with MHT. PMID- 10678550 TI - Relationship of renal histological damage to glomerular hypertension in patients with immunoglobulin A nephropathy. AB - OBJECTIVE: Studies of experimental animals show glomerular hypertension to be important in the progression of glomerular disease. We evaluated this connection clinically by examining the relationship between glomerular hemodynamics and histological changes in patients with immunoglobulin (Ig)A nephropathy. METHODS: The subjects were 23 patients with IgA nephropathy. All patients underwent renal biopsies. Glomerular hemodynamics, in terms of glomerular capillary hydraulic pressure (PGC) and the whole-kidney ultrafiltration coefficient, were calculated from the renal clearance, plasma total protein concentration, and pressure natriuresis relationship. The severity of glomerulosclerosis, tubulointerstitial damage and mesangial matrix expansion was evaluated semiquantitatively. RESULTS: PGC ranged from 33-69 mm Hg, and the mean arterial pressure (MAP) from 79-112 mm Hg. Their correlation was not significant (r= 0.29, P= 0.18). PGC was significantly correlated with the glomerulosclerosis score, and also with the score for tubulointerstitial damage (r= 0.65, P < 0.001 and rs = 0.59, P = 0.007, respectively), but not with the score for mesangial matrix expansion (r= 0.08, P= 0.72). MAP was significantly correlated only with the score for tubulointerstitial damage (rs = 0.63, P = 0.004). In multiple linear regression analysis of the histological changes and hemodynamics, the glomerulosclerosis score and the score for tubulointerstitial damage were correlated with PGC, but not with MAP. CONCLUSION: These clinical results support the speculation that glomerular hypertension is involved in the glomerulosclerosis and tubulointerstitial damage that occurs in IgA nephropathy. PMID- 10678551 TI - Development of different phenotypes of hypertensive heart failure: systolic versus diastolic failure in Dahl salt-sensitive rats. AB - OBJECTIVE: There are two phenotypes of heart failure, systolic failure and isolated diastolic heart failure with preserved left ventricular systolic function. Although isolated diastolic heart failure frequently occurs, there are only models for diastolic dysfunction unassociated with heart failure and models with overt diastolic heart failure have not been established. We attempted to develop two different models, i.e. diastolic and systolic failure models, based on hypertension. MATERIALS AND METHODS: Dahl salt-sensitive rats were placed on 8% NaCl diet from 7 weeks old (7-week starting group) or 8 weeks old (8-week starting group). As an age-matched control, Dahl salt-sensitive rats were consistently placed on normal chow. In these rats, echocardiogram was serially recorded, followed by hemodynamic and histological studies. RESULTS: The 7-week starting rats showed a steep elevation in blood pressure and progressive left ventricular hypertrophy, and fell into overt heart failure at approximately 19 weeks. The development of heart failure was not associated with a decrease in left ventricular midwall fractional shortening or an increase in left ventricular end-diastolic dimension as compared with the age-matched control, which mimics the characteristics of clinically observed isolated diastolic heart failure. The 8-week starting rats showed a gradual rise in blood pressure and less progressive left ventricular hypertrophy, and fell into heart failure at approximately 26 weeks with a decrease in mid-wall fractional shortening and an increase in left ventricular end-diastolic dimension. Hemodynamic and histological studies at failing stage revealed comparable elevation of left ventricular end-diastolic pressure and comparable left ventricular fibrosis in both groups. CONCLUSION: These two different models of overt heart failure may be useful as models of isolated diastolic heart failure and systolic heart failure based on the same hypertensive heart disease, respectively, and may contribute to discrimination of the mechanisms of the development of the two different phenotypes of heart failure. PMID- 10678552 TI - How to assess endothelial function in human blood vessels. PMID- 10678553 TI - Tumor markers in monitoring response to chemotherapy for patients with gastric cancer. PMID- 10678555 TI - Frequency of HLA-A alleles in Japanese patients with head and neck cancer. AB - BACKGROUND: Association between certain human leukocyte antigen (HLA) types such as HLA-A1 and -A3 and squamous cell carcinoma of the head and neck (SCCHN) has been demonstrated in the Caucasian population. HLA typings in these studies were performed by conventional serological methods. However, recent comparison studies between serological and molecular typings have revealed that the former are often inaccurate. METHODS: The frequency of HLA-A alleles in 100 Japanese patients with SCCHN and 100 control subjects was determined by the polymerase chain reaction, with primers specific for the HLA-A locus, in combination with dot-blot hybridization with 31 sequence-specific oligonucleotides. RESULTS: The frequencies of HLA-A*2602 and HLA-A*3303 were higher and those of HLA-A*2603 and HLA-A*3101 were lower in the patients with SCCHN than in healthy controls, but these differences were not statistically significant. In the 39 male patients with laryngeal carcinoma, the most common malignancies in Japanese patients with SCCHN, the frequency of HLA-A*2402 was significantly lower than that in the 80 male controls; however, after correction of the P value, statistical significance was not confirmed. In oral carcinoma patients, the frequency of HLA-A*2402 was significantly higher than that in healthy controls. CONCLUSIONS: These results suggest that the contribution of certain HLA-A alleles to susceptibility to SCCHN may differ between sites in the head and neck regions, despite these cancers being of an identical histological type, and that HLA-A*2402 may influence the development of oral carcinoma in Japanese patients. PMID- 10678554 TI - Expression pattern of chemoresistance-related genes in human malignant brain tumors: a working knowledge for proper selection of anticancer drugs. AB - BACKGROUND: In addition to traditional modalities such as surgical intervention and radiotherapy, chemotherapy is a common therapeutic method for human malignant brain tumors. However, the effectiveness of chemotherapy is frequently hampered by cancer cell chemoresistance, resulting in an unsatisfactory outcome. To overcome this disadvantage, the proper selection of efficacious anticancer agents is required. METHODS: The expression levels of chemoresistance-related genes, MGMT, mdr1, MRP, MTIIA and GST-pi, in 28 surgical specimens of human brain tumors and in 10 human glioma cell lines were examined by Northern blot analysis. In addition, the SD10 values of human glioma cell lines against ACNU, CDDP, ADM and VP16 were estimated by a cell survival assay. RESULTS: The expression levels of each of the chemoresistance-related genes, except MRP, were generally higher in brain tumors than those in non-neoplastic brain tissues. MGMT expression correlated exclusively with ACNU resistance in all glioma cell lines examined (p = 0.0002). The transcriptional level of mdr1 in the tumor cells correlated with the SD10 values of VCR (p = 0.04) and ADM (p = 0.034). In contrast, the expression levels of MTIIA and GST-pi did not correlate with resistance to any of the drugs tested. A correlation of MRP mRNA expression with multidrug resistance was not apparent in the 10 cell lines tested. CONCLUSIONS: The data indicate that knowledge of the expression levels of MGMT and mdr1 may be particularly useful for a more rational selection of drugs which are not influenced by these resistance genes and which have improved efficacy against human brain tumors. PMID- 10678556 TI - Lung cancer in patients who had received thoracoplasty for pulmonary tuberculosis. AB - BACKGROUND: In Japan in the 1950s, thoracoplasty was a powerful therapy for pulmonary tuberculosis. Now there are many aged people who have tuberculosis sequelae caused by thoracoplasty. We have encountered some cases of lung cancer among these people. METHODS: To elucidate the features of lung cancer occurring after thoracoplasty for pulmonary tuberculosis, we reviewed for analysis 20 such cases. RESULTS: There were 17 men and three women, aged 55 to 78 years (mean 65 years). All had respiratory dysfunction and most were cigarette smokers. Lung cancers were located in the upper lobes in nine cases, in the middle lobe in one and in the lower lobes in 10. Ten lung cancers were in the thoracoplastied lung and the remaining 10 in the opposite lung. Histologically, squamous cell carcinoma was predominant (11 cases). Eight lung cancers were detected in stages I and II and 12 in stages III and IV. Most cancer lesions were separate from tuberculosis lesions. Surgical resection was selected in only three of 11 stages I-IIIA cases in consideration of respiratory dysfunction and/or ventilatory impairment due to thoracoplasty. Chemotherapy and/or radiotherapy were performed in nine and supportive care alone was performed in eight. Fourteen patients died of lung cancer and four died of cor pulmonale due to tuberculosis sequelae. Five year survival was achieved in only one surgical case. Eight of the nine patients who received chemotherapy or radiotherapy died within 1 year, and, further, seven of eight patients who received supportive care died within 6 months. CONCLUSION: Lung cancer in the patients who had received thoracoplasty occurred in each lung and every lobe, independent of thoracoplasty. In addition, delay of detection was such that stage III-IV cases were in the majority, there were some limitations in therapeutic benefits related to thoracoplasty and the prognosis was very poor. Physicians should avoid delay in the detection of lung cancer through careful follow-up of such patients. PMID- 10678557 TI - Phase I study of cisplatin and docetaxel plus mitomycin C in patients with metastatic non-small cell lung cancer. AB - BACKGROUND: Docetaxel, cisplatin and mitomycin C are some of the active drugs used in the treatment of patients with metastatic non-small cell lung cancer (NSCLC). The purpose of this study was to determine the maximum tolerated dose (MTD) and recommended dose of the three drugs in combination for such patients. METHODS: Chemotherapy-native patients with metastatic NSCLC were enrolled in this study. The doses of docetaxel and cisplatin were fixed at 60 and 80 mg/m2, respectively. It was planned to increase the dose of mitomycin C from 4 to 6 and 8 mg/m2. All drugs were administered on day 1 and repeated every 3-4 weeks. RESULTS: All six patients received 60 mg/m2 of docetaxel and 80 mg/m2 of cisplatin, three of them with 4 mg/m2 of mitomycin C (level 1) and the other three with 6 mg/m2 of mitomycin C (level 2). Two of the three level 2 patients experienced dose-limiting toxicities (DLTs) in first cycle: febrile neutropenia and grade 3 hyponatremia. Based on these data, the MTD was concluded to be 60 mg/m2 for docetaxel, 80 mg/m2 for cisplatin and 6 mg/m2 for mitomycin C. Evaluation of the data from all of the cycles, however, showed that four of the six patients experienced DLTs. CONCLUSIONS: The addition of mitomycin C to docetaxel and cisplatin resulted in relatively high toxicities. It was impossible to use a high enough dose of mitomycin C to improve the survival of NSCLC patients. We therefore concluded that further evaluation of this combination is unwarranted. PMID- 10678558 TI - Tumor markers CEA, CA19-9 and CA125 in monitoring of response to systemic chemotherapy in patients with advanced gastric cancer. AB - BACKGROUND: To evaluate whether tumor markers can be used to assess response to systemic chemotherapy, we analyzed preliminarily the relationship between the response to chemotherapy based on serial imaging and on change in serum tumor marker level of CEA, CA19-9 and CA125. METHODS: We analyzed 26 patients with advanced gastric cancer in whom at least one of the tumor markers CEA, CA19-9 and CA125 was elevated before systemic chemotherapy with regard to the relationship between the change in serum tumor marker level and response assessment by imaging studies throughout the treatment course. A responder was defined as showing a > or = 50% drop in tumor marker level for more than 4 weeks. RESULTS: The sensitivity and negative predictive value of falling tumor marker level after chemotherapy for a partial response in imaging was 100%. When patients were categorized as responders or non-responders, a significant correlation was observed between the assessment of response by tumor markers and by imaging studies. The survival time of responders assessed by tumor markers was significantly longer than that of non-responders. CONCLUSIONS: The measurement of tumor markers might be useful in monitoring response and in predicting the prognosis of patients with advanced gastric cancer treated with systemic chemotherapy. Tumor markers may be used as a means of monitoring treatment in patients when in an imaging study it is difficult to assess response to chemotherapy in clinical practice. Further studies are required to confirm these findings. PMID- 10678559 TI - Clinicopathological comparisons of familial and sporadic cases in 219 consecutive Japanese epithelial ovarian cancer patients. AB - BACKGROUND: It is estimated that approximately 5-10% of epithelial ovarian cancer patients in Western countries are associated with an autosomal dominant inheritance with variable penetrance. There are a few reports of familial ovarian cancer in Japan and considerable uncertainties remain regarding Japanese familial ovarian cancer. The aim of this study was to clarify the clinicopathological features of Japanese familial ovarian cancer. METHODS: We studied clinicopathological findings for 219 consecutive epithelial ovarian cancer patients treated at our institution from April 1987 to September 1997. RESULTS: Eleven patients in nine families were diagnosed as familial ovarian cancer and the incidence of familial cases was 5.0%. Most women (90.9%) with familial cases were diagnosed as the breast ovarian cancer syndrome, whereas ovarian cancer associated with hereditary nonpolyposis colorectal cancer was relatively rare (9.1%). Serous adenocarcinoma, high histological grade, advanced FIGO stage and breast cancer as multiple primary cancer were significantly more common in familial cases compared with sporadic cases (p < 0.001, p < 0.05, p < 0.005 and p < 0.005, respectively). Earlier age of onset was thought to be a characteristic of familial ovarian cancer in Western countries; however, we did not find any difference in age at diagnosis between familial and sporadic cases (53.4 vs 51.3 years). The prognosis of familial ovarian cancer remains controversial and our data did not show a significant difference (p = 0.45) in prognosis between these two groups. CONCLUSION: These findings, except for age at diagnosis, in Japanese familial ovarian cancer are in accordance with the features of familial ovarian cancer in Western countries. PMID- 10678561 TI - Surgical management of primary lung cancer in an elderly patient with preoperative empyema. AB - A 74-year-old man with primary lung cancer developed preoperative empyema but was successfully managed surgically. The patient was given a diagnosis of c-T2N1M0, stage IIB, moderately differentiated squamous cell carcinoma, but before surgery pneumothorax and empyema developed, resulting from rupture of the carcinoma. Thoracic drainage, lavage and systemic administration of antibiotics improved his empyema. As there were no malignant cells in the drainage fluid, right middle lower bilobectomy, empyemal cavity resection and lymph node dissection were performed. The bronchial stump was covered with an intercostal muscle flap. Thoracic drainage, lavage and systemic administration of antibiotics were performed for 6 days following the operation. The patient was discharged on the 27th postoperative day without any complications having developed. The pathological diagnosis of the tumor was p-T4N2(#7)M0, stage IIIB, br(-), ly(+), v(+), p3(pleura), pm1 and d0. He died of recurrence at home 18 months after the operation. We believe the following to be the minimum requirements for surgical management of such patients: (1) immediate thoracic cavity drainage and lavage with systemic antibiotic therapy, aiming at infection control before surgery; (2) prophylactic lavage of the thoracic cavity during and after surgery and (3) coverage of the bronchial stump with an adequate flap. Six reported cases of primary lung cancer with preoperative empyema are also discussed. PMID- 10678560 TI - Goserelin acetate with or without antiandrogen or estrogen in the treatment of patients with advanced prostate cancer: a multicenter, randomized, controlled trial in Japan. Zoladex Study Group. AB - OBJECTIVE: The aims of this randomized, controlled study were to investigate the efficacy and safety of long-term monotherapy with the luteinizing hormone releasing hormone agonist goserelin acetate compared with both short- and long term combined androgen blockade. METHODS: Patients with advanced prostate cancer (n = 371) were randomized to treatment with goserelin acetate alone or a combination of goserelin acetate plus either long-term or short-term antiandrogen (chlormadinone acetate) or short-term estrogen (diethylstilbestrol diphosphate). RESULTS: There were no significant differences between the treatment groups with respect to objective progression, overall survival or disease-specific survival. Nevertheless, subgroup analysis suggested that patients with minimal disease or a good prognosis might benefit more from combined androgen blockade than other patients. Combined androgen blockade significantly reduced the incidence of disease flare compared with goserelin acetate treatment alone. CONCLUSIONS: Neither short- nor long-term combined androgen blockade had a survival advantage over goserelin acetate alone. PMID- 10678562 TI - Possible associations of rectal carcinoma with Schistosoma japonicum infection and membranous nephropathy: a case report with a review. AB - We report the first case of rectal carcinoma associated with S. japonicum and membranous nephropathy. A 57-year-old Japanese man noticed narrowing of his feces. He had lived in Yamanashi prefecture, an endemic area of S. japonicum. He had suffered from nephrotic syndrome for about 1 year. Barium enema study showed a severe stricture in the upper rectum and biopsy specimens from the tumor demonstrated well differentiated adenocarcinoma and many ova of S. japonicum. Sonography of the liver showed a network pattern and a linear high echoic area. Low anterior resection with incisional biopsy of the liver and the right kidney was performed. Histopathological findings showed well differentiated adenocarcinoma and schistosomal ova. The total number of ova in the resected colon amounted to 15,133, consisting of 2243 inside and 12,890 outside the carcinoma. The nearer to the carcinoma the area was, the higher was the density of ova. The findings of light microscopy and electron microscopy of the biopsy specimen from the kidney were compatible with membranous nephropathy (stage II). This case suggests that schistosomal ova have some effect on carcinogenesis and nephrotic syndrome. In patients with nephrotic syndrome of unknown cause, especially in inhabitants of endemic areas of S. japonicum, gastrointestinal malignancy should be ruled out as an etiological factor. Sigmoidoscopy would be useful for colorectal carcinoma surveillance in S. japonicum patients. PMID- 10678563 TI - Cisplatin-5-fluorouracil therapy with remarkable effect and 5-year survival for paraaortic lymph node metastases of rectal carcinoma in females: a case report. AB - A 68-year-old woman was admitted because of a rectal carcinoma with huge paraaortic lymph node metastases. Low anterior resection with regional lymph node dissection was performed, leaving the paraaortic mass. After the operation, cisplatin-5-fluorouracil therapy was used as supplemental chemotherapy. The metastatic lymph nodes shrank remarkably in response to anticancer drugs. We evaluated the effect of chemotherapy as a partial response. The physical condition of the patient was well controlled for more than 4 years until she was admitted again because of cardiac failure accompanied by relapse of abdominal lymph node swelling. She died of cardiac failure 5 years and 3 days after the operation. PMID- 10678564 TI - The probability of a Japanese person developing cancer during their lifetime. PMID- 10678565 TI - Gene regulation during the development of embryonic vascular endothelium- reflections of an Irish postdoc in Japan. PMID- 10678566 TI - Subtle signs may mask cancer-related emergencies. PMID- 10678567 TI - Cancer statistics digest. Comparison of mortality and incidence trends in Japan. PMID- 10678568 TI - International Agency for Research on Cancer (http://www.iarc.fr/) PMID- 10678569 TI - Phenotypical and morphological alterations to rat sinusoidal endothelial cells in arterialized livers after portal branch ligation. AB - The hepatic sinusoids are preferentially supplied with portal venous blood and equipped with fenestrated endothelial cells that are distinct from capillary endothelial cells. We previously observed in rats that sinusoidal capillarization proceeded concurrently with arterial blood supply during hepatocarcinogenesis. This study aimed to clarify the inducing role of arterialization in sinusoidal capillarization by investigating phenotypical, morphological and functional alterations to sinusoidal endothelial cells (SECs) in arterialized rat livers induced by portal branch ligation. At one week, after massive hepatic necrosis following ligation, the livers were restored to their normal architecture without causing post-necrotic fibrosis. At 12-21 weeks, they exhibited a normal histology except for mild pericellular fibrosis which developed along sinusoids or between adjacent hepatocytes. SECs expressed factor VIII-related antigen and showed a decrease in the number of fenestrae and porosity, still lacking any basement membrane but further retaining the functional capacity for carmine dye uptake. Stellate cells, while occasionally associated with large amounts of collagen bundles, contained many lipid droplets and expressed no alpha-smooth muscle actin, indicating a quiescent property. Kupffer cells were commonly found within the sinusoids. The present results indicate that arterialization of the liver induces a partial (but not complete) transition of SECs into capillary-type endothelial cells, suggesting that arterialization might be one of the factors which induce sinusoidal capillarization in the development of hepatocellular carcinoma. PMID- 10678570 TI - The role of Kupffer cells in liver regeneration. AB - The liver has a remarkable proliferative capacity after a partial hepatectomy. Previous studies have indicated that Kupffer cells have the potential to exert both stimulatory and inhibitory influences on hepatocyte proliferation. To elucidate the role of Kupffer cells in liver regeneration, mice were selectively depleted of Kupffer cells by injection of liposome-encapsulated dichloromethylene diphosphonate (lipo-MDP) at day 3 after a two-thirds hepatectomy. Results showed that liver regeneration was delayed after Kupffer cell-depletion. In control mice, hepatocyte growth factor (HGF) mRNA expressions were enhanced during liver regeneration and expressions of HGF were localized in fat-storing cells (Ito cells). In Kupffer cell-depleted mice, the number of HGF-expressing cells decreased in the regenerating liver, and expressions of HGF and its receptor (c met) as well as other growth factors/cytokines were less prominent than in control mice. In contrast, expressions of TNF-alpha, another potent cytokine involved in liver regeneration, did not differ between Kupffer cell-depleted and control mice during the regeneration. Administration of TNF-alpha antibody did not reduce the expression of HGF or liver regeneration. These findings imply that Kupffer cells play a stimulatory role in liver regeneration by enhancing HGF expression via TNF-alpha-non-mediated mechanisms. PMID- 10678571 TI - The induction of autophagic vacuoles and the unique endocytic compartments, C shaped multivesicular bodies, in GH4C1 cells after treatment with 17beta estradiol, insulin and EGF. AB - The mechanisms for the formation of autophagic vacuoles were investigated using GH4C1 cells, a rat pituitary tumor cell line, whose induction increases intracellular levels of lysosomal proteinases and their mRNA by treatment with a combination of hormones (17beta-estradiol, insulin and EGF). By ordinary electron microscopy, autophagic vacuoles containing various undigested structures with or without limiting membranes were abundant in the hormone-induced cells. These vacuoles, also containing numerous small vesicles, appeared to be derived from multivesicular bodies. In fact, there were also numerous C-shaped multivesicular bodies which enclosed cytoplasmic portions, suggesting that these unique structures are involved in the production of the autophagic vacuoles. Moreover, the cytoplasmic portions enlapped by the C-shaped multivesicular bodies were high in electron density and contained filamentous structures. By the cryothin-section immunogold method, the C-shaped multivesicular bodies in some cases contained lysosomal marker proteins such as cathepsins B and H, and Igp 120. Using an anti actin monoclonal antibody, immunogold particles clearly labeled the cytoplasmic portions enclosed by the C-shaped multivesicular bodies. Pulse-chase experiments with horse radish peroxidase, a fluid-phase endocytic marker, revealed that the incidence of the C-shaped multivesicular bodies labeled with horse radish peroxidase peaked at 30 min after the beginning of chase incubation, whereas no C shaped multivesicular body with horse radish peroxidase was detected in the cells by cytochalasin D treatment. These results suggest that the C-shaped multivesicular bodies occur in a transitional process from endosomes to lysosomes by the action of actin filaments, and that this morphological change may be essential for the production of autophagic vacuoles in the hormone-induced GH4C1 cells. PMID- 10678572 TI - Heterogeneous localizations of Trk B among individual periodontal Ruffini endings in the rat incisor. AB - The present immunocytochemical study examined the localization of Trk B, a high affinity neurotrophin receptor, in the neural elements of the periodontal ligament of the rat incisor. In light microscopy, the immunoreactivity was demonstrated in dendritic profiles in the alveolar half of the periodontal ligament. Their location and morphological features indicated that they were periodontal Ruffini endings. Occasional rounded cells associated with periodontal Ruffini endings, which had immunonegative kidney-shaped nuclei, were immunoreactive; these were judged to be terminal Schwann cells. Immunoelectron microscopy revealed the heterogeneous localization of Trk B among individual Ruffini endings. Some terminal Schwann cells contained immunoreactive products for Trk B in the cytoplasm, while others did not. Similarly, a part of the Schwann sheaths covering the axon terminals showed Trk B immunoreactivity. Most axon terminals associated with periodontal Ruffini endings were immunopositive for Trk B, though a few of them were immunonegative. The ordinary Schwann cells did not contain Trk B immunoreactive products. These findings imply that Trk B is required for the maintenance of periodontal Ruffini endings. The different expression pattern of Trk B suggests that neuronal and glial elements comprising individual periodontal Ruffini endings are subject to heterogeneous conditions with regard to the requirement of Trk B. PMID- 10678573 TI - Immunocytochemical localization of taurine in the developing retina of the lefteye flounder Paralichthys olivaceus. AB - Light microscopic immunolocalization of taurine, a sulfur-containing free amino acid, was investigated in the developing retina of a lefteye flounder, Paralichthys olivaceus, which exhibits metamorphic changes with rod cell addition for 3-5 weeks after hatching. This immunocytochemical study of the developing retina revealed: 1) From 3 to 13 days after hatching, intense immunostaining was shifted from the surroundings of neural cells to the neural somata and processes in the inner retina. 2) Intense immunoreactivity appeared also in the outer and inner segments and basal processes (pedicles) of cone cells within 6 days or 13 days after hatching. 3) Lack of immunoreactivity was found in the outer segment of rod cells from their appearance during metamorphosis. These findings are discussed with the possible functional roles of taurine in the fish retina: 1) involvement in cell differentiation and/or development; 2) protection of the outer segments against light stimuli; and 3) regulation of neural transmission. PMID- 10678574 TI - Reappraisal of potassium permanganate oxidation applied to Lowicryl K4M embedded tissues processed by high pressure freezing/freeze substitution, with special reference to differential staining of the zymogen granules of rat gastric chief cells. AB - The high pressure freezing/freeze substitution technique is known to yield a deep vitreous freezing of tissues. Combination of this technique with Lowicryl K4M embedding allows us histochemical studies of dynamic cellular processes with improved structural preservation. The disadvantage of Lowicryl K4M embedding is its poor electron density in electron microscopy. To address this problem, we examined the effects of KMnO4 oxidation applied to Lowicryl K4M embedded rat gastric glands processed by high pressure freezing. The KMnO4 oxidation-uranyl acetate-lead citrate sequence succeeded not only in contrast enhancement of cellular components, but also in differential staining of the zymogen granules of rat gastric chief cells. This technique could be applied to semi-thin sections of Lowicryl K4M embedded rat gastric glands. The KMnO4 oxidation-toluidine blue staining provided sufficient contrast with regard to the zymogen granules. Various experiments used in this study verified that the KMnO4 oxidation plays an essential role in the differential staining of the zymogen granules. Combined use of the KMnO4 oxidation with phospholipase A2-immunostaining demonstrated that gold labeling was localized to the zymogen granules without the loss of immunolabeling. Energy dispersive X-ray microanalysis revealed some manganese depositions on the zymogen granules. It is highly anticipated that the KMnO4 oxidation will become a useful tool for histochemical investigations combined with cryofixation/freeze substitution and low temperature embedding techniques. PMID- 10678575 TI - Immunolocalization of tight junction proteins, occludin and ZO-1, and glucose transporter GLUT1 in the cells of the blood-nerve barrier. AB - Facilitated-diffusion glucose transporter GLUT1 is abundant in the blood-nerve barrier. To observe the relationship between glucose transfer across the barrier and the molecular architecture of the barrier, we examined the localization of GLUT1 and tight junction proteins, occludin and zonula occludens-1 (ZO-1), by immunofluorescence microscopy and immunogold electron microscopy in the rat sciatic nerve. GLUT1 was enriched at the whole aspects of the plasma membranes of the cells of the barrier: perineurial cells, and endothelial cells of the blood vessels in the endoneurium. These GLUT1-positive cells were also positive for occludin and ZO-1, both of which were localized at tight junctions. ZO-1 additionally was present in the GLUT1-negative cells not serving as the blood nerve barrier. These observations suggest that occludin in the tight junctions and GLUT1 at the plasma membranes in the cells of the barrier may constitute a mechanism for the selective transfer of glucose across the barrier while preventing the non-specific flow of blood constituents. PMID- 10678576 TI - Porosity of the epithelial basement membrane as an indicator of macrophage enterocyte interaction in the intestinal mucosa. AB - The epithelial basement membrane of intestinal villi is perforated with numerous small pores, through which free cells in the lamina propria communicate with the enterocytes. This study was a comparative analysis of the pores in the basement membrane by SEM after removal of the gut epithelium with OsO4 maceration. The porosity as represented by the area fraction of the pores varied along the baso apical axis of villi in patterns specific for each animal species examined: consistent scantiness along the entire length of villi in mice, acute elevation in the second and third distal one-sixths of villi in rats, and gradual augmentation toward the villus tips in guinea pigs. Size distribution analyses of the pores indicated their heterogeneous enlargement in the regions of elevated porosity. Concomitant observation of lamina propria macrophages by histochemical labelings and by conventional TEM showed that the cells specifically clustered beneath the hyperporous basement membrane, with their thick processes penetrating it. The spatially-regulated patterns of perforation of the epithelial basement membrane indicate phase-specific interventions of lamina propria macrophages in the maturation or aging of enterocytes, which steadily proliferate in crypts and exfoliate at the villus tips. PMID- 10678577 TI - Immunocytochemical demonstration of heat shock protein 25 in the rat temporomandibular joint. AB - The expression of heat shock protein 25 (Hsp 25) was investigated in the rat temporomandibular joint by immunocytochemistry combined with confocal and electron microscopy. Immunostaining with an antibody to Hsp25 was able to demonstrate various cellular elements in the synovial membrane of the joint. Intense immunoreaction for Hsp25 was recognized in certain cells comprising the synovial lining layer. Confocal microscopic observation revealed two characteristic profiles of the Hsp25-positive cells with cytoplasmic processes: one extended thick and long processes towards the articular cavity, and the other prejected horizontally slender processes which covered the synovial membrane. Under the electron microscope, the immunoreactive synovial lining cells were characterized by a well-developed rough endoplasmic reticulum and secretory granules, suggesting that they can be categorized as fibroblastic type B cells. The covering by the cytoplasmic extensions was confirmed by immuno-electron microscopic observations. This cytoplasmic covering presumably performs a barrier function and expedites the effective secretion/resorption of synovial fluids. Since it has been proposed that Hsp 25 is associated with an estrogen receptor, the immunopositive synovial lining cells were considered estrogen-target cells. Immunoreactivity for Hsp25 was also observed in the chondrocytes of the maturative and hypertrophic cell layers as well as in the cells of the articular disk. A suggestion was made that Hsp25 might be involved in the inhibition of apoptosis of those cells. PMID- 10678578 TI - Microdissection or microspot CO2 laser for limited vocal fold benign lesions: a prospective randomized trial. AB - CO2 lasers have become an important technological advance and an integral tool for the laryngeal surgeon since the 1960s. Surgeons have used lasers for a variety of benign and malignant lesions in the larynx with good success. With better understanding of the microarchitecture of the vocal folds and the recognition of heat distribution into surrounding tissues that occurs with the use of standard CO2 lasers, questions and concerns have been raised regarding the use of the CO2 laser for benign lesions of the vocal folds. With the advent of the microspot CO2 laser with a spot size of less than 250 microm, the potential heat distribution to the deeper layers of the lamina propria has been reduced. The microspot CO2 laser has been suggested to be an appropriate tool for the excision of superficial benign lesions of the vocal fold and may be considered as an appropriate treatment alternative to microdissection. Only a limited number of studies have compared the efficacy of microdissection versus microspot CO2 laser surgery in the larynx, and no prospective, randomized trials have been performed. OBJECTIVE: This study was designed to compare microspot CO2 laser excision and microdissection for superficial benign lesions confined to the free margin of the vocal fold. STUDY DESIGN: A randomized, prospective trial comparing microspot CO2 laser excision and microdissection in the removal of nodules, polyps, and mucous retention cysts of the vocal fold. METHODS: Acoustic and aerodynamic measures and videostroboscopic and perceptual audio recordings evaluated by a panel of blinded viewers and listeners were studied preoperatively and 2 to 3 weeks and 5 to 12 weeks postoperatively. Surgical and recovery times were compared between the two groups. RESULTS: Thirty-seven patients met selection criteria and were enrolled, 21 in the microdissection group and 16 in the laser excision group. Significant improvements in videostroboscopic parameters were found over time in both groups. Significant improvements were noted for perceptual analysis over time for the laser excision group with nonsignificant improvements over time for the microdissection group. There was no difference in any measure between laser excision and microdissection at the two postoperative visits. There was no difference in surgical or recovery time between laser excision and microdissection. Acoustic and aerodynamic parameters were noncontributory in evaluating outcomes of treatment, since most values were normal before surgery. CONCLUSION: No differences in clinical outcomes are identified when comparing microdissection with laser excision of nodules, polyps, and mucous retention cysts of the vocal folds. PMID- 10678579 TI - Molecular pathology of cyclooxygenase-2 in neoplasia. AB - Cyclooxygenase (COX)-2 levels are elevated in several types of human cancer tissues. Nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit both the COX-1 and COX-2 protein, the two enzymes that convert arachidonic acids to prostaglandins. Regular use of such NSAIDs significantly reduces the risk and spread of some cancers. The objective of this study was to elucidate the molecular pathology of neoplasms that overexpress COX-2. Epidemiological data and clinical studies were analyzed and compared with results of studies of human tumor tissues, animal models, and cultured tumor cells. COX-2, but not COX-1, is highly expressed in human colon carcinoma, squamous cell carcinoma of the esophagus, and skin cancer. COX-2 is inducible by oncogenes ras and scr, interleukin-1, hypoxia, benzo[a]pyrene, ultraviolet light, epidermal growth factor, transforming growth factor beta, and tumor necrosis factor alpha. Dexamethasone, antioxidants, and tumor-suppressor protein p53 suppress COX-2 expression. COX-2 synthesizes prostaglandin E2 (PGE2) which stimulates bcl-2 and inhibits apoptosis, and induces interleukin-6 (IL-6) which enhances haptoglobin synthesis. PGE2 is associated with tumor metastases, IL-6 with cancer cell invasion, and haptoglobin with implantation and angiogenesis. Drastic reduction in polyp number results from COX-2 gene knockout as well as from selective COX-2 inhibition in a mouse model of human familial adenomatous polyposis. Nonselective NSAIDs, for instance aspirin, and selective COX-2 inhibitors such as celecoxib (SC-58635) and NS-398 suppress azoxymethane-induced colon carcinogenesis in rats. Aspirin, indomethacin, and ibuprofen decrease cultured lung cancer cell proliferation. Selective inhibition of COX-2 is preferable to nonselective inhibition. It reduces cancer cell proliferation, induces cancer cell apoptosis, and spares COX-1-induced cytoprotection of the gastrointestinal tract. PMID- 10678580 TI - Wellness assessment: A role for laboratory medicine. AB - Within the next 20-25 years, 20% of the U.S. population is expected to be 65 years old or older. Unless significant changes are made (ie, increased research, improved treatment, and promotion of health and disease prevention), medical costs will continue to rise but with fewer citizens to finance them. However, this need not be the case. The 10 most common causes of death in the U.S. are all, to a significant degree, lifestyle-related. Most of these diseases take years or even decades to develop. By the time a physician makes a diagnosis, the disease is already present. However, with a panel of laboratory tests, combined with age, sex, family history, body mass index, blood pressure, and information regarding lifestyle (diet, exercise, smoking, alcohol), an estimate can be made regarding the relative risk for developing the major diseases/disorders. If recommendations are followed, these conditions will decline, medical costs will be reduced, and people will enjoy longer, healthier, and more productive lives. PMID- 10678581 TI - Molecular predictive factors for local recurrence and distant metastasis of breast cancer after lumpectomy with postoperative radiation therapy. AB - To determine the risk factors associated with the recurrence and metastasis of breast cancer after lumpectomy with postoperative radiation therapy, 112 cases were studied who had been treated during a period of 11 years at the University of Florida Health Science Center/Jacksonville. The patients were evaluated for their age, race, and clinical stage, as well as the tumor grade, stage, histological type, and node involvement. Among these cases, four (4%) recurred locally within a year of treatment; 10 (9%) cases presented with distant metastasis within three years. No obvious clinical risk factors were identified for local recurrence; however, positive-node status seemed to be associated with distant metastasis. The primary tumors of these cases were then studied using immunohistochemical staining to evaluate the potential prognostic value of tumor markers such as estrogen receptor (ER), progesterone receptor (PR), tumor suppressor gene p53, HER-2/neu oncogene, and multi-drug resistance gene (MDR). The expression of p53 was associated with all local recurrence cases as well as 50% of those who had metastasis. The expression of MDR was observed in 80% of the distant metastatic cases. This preliminary result may warrant further studies on larger number of cases to assess the predictive value of p53 and MDR in the outcome of breast cancers in patients treated with postoperative radiation therapy. PMID- 10678582 TI - Fluorescence in situ hybridization (FISH) for detection of HER-2/neu amplification in breast cancer: a multicenter portability study. AB - Amplification and/or overexpression of HER-2/neu has been shown to be both a prognostic and predictive marker in breast cancer. Recent studies have also confirmed the efficacy of Herceptin (trastuzumab) as adjuvant therapy for patients with overexpression of HER-2/neu. Therefore, it is critical that precise and reproducible assays be used in the clinical laboratory setting for determination of the HER-2/neu status in patients with breast cancer. The objective of this study was to determine the portability (reproducibility between different institutions) of the PathVysion HER-2 fluorescence in situ hybridization (FISH) assay used for detection of amplification of the HER-2/neu gene in formalin-fixed, paraffin-embedded tissue sections of invasive ductal carcinoma of the breast. Study specimens consisted of one breast tumor with a normal HER-2/neu copy number, two tumors with a low level, and one tumor with a high level of HER-2/neu amplification. The PathVysion HER-2 assay was shown to be highly reproducible on different assay days (n = 3) and between different institutions (n = 5) in the detection of amplification of the HER-2/neu gene in routinely processed clinical specimens of breast carcinoma. In addition, this study examined the feasibility of enumerating FISH signals in 20 nuclei in contrast to 60 nuclei per specimen. Although a modest increase in variation was observed when analyzing 20 compared to 60 nuclei, the mean ratios were similar. Therefore, analysis of as few as 20 nuclei with this FISH HER-2/neu assay may be sufficient for determining the amplification level of the HER-2/neu gene. PMID- 10678583 TI - Quantification of HER2 oncoprotein in fine-needle aspirates of the breast. AB - Measurements of either HER2 gene overexpression or its gene-coded protein (p185) are clinically useful for predicting prognosis in breast cancer. The measurements are also useful for identifying metastatic breast cancer patients who may benefit from Herceptin treatment. Since fine needle aspiration (FNA) of the breast has become an increasingly popular technique for obtaining tissue specimens, we have developed a sensitive method to quantify p185 in the aspirate. For this procedure, p185 from the cell pellet of FNA is extracted with a buffer containing Triton X-100, and the p185 is measured with an enzyme immunoassay. Most of the malignant breast tumors (N=7) in this study were associated with elevated p185 concentrations (6/7, 319+/-222 U/mg), compared to the p185 concentrations in normal breast tissue (42.8+/-35 U/mg, N=47) or benign lesions (43.1+/-20.2 U/mg, N=22). Quantification of p185 in FNA may improve the assessment of breast cancer patients, revealing whether they are at high risk and may benefit from Herceptin treatment. PMID- 10678584 TI - Glutathione-S-Transferase as a selective inhibitor of oncogenic ras-p21-induced mitogenic signaling through blockade of activation of jun by jun-N-terminal kinase. AB - We have identified the intracellular detoxification enzyme, glutathione-S transferase (GST), as a potent inhibitor of the activation of jun by its kinase, jun-N-terminal kinase (JNK), in vitro. All three major isozymes (alpha, mu, and pi) bind to JNK-jun complexes and inhibit activation of jun by JNK. We now find that GST inhibits JNK-induced oocyte maturation in vivo and strongly inhibits oocyte maturation induced by oncogenic ras-p21 protein, but not by insulin activated normal cellular p21 protein. These results correlate with the finding that oncogenic, but not insulin-activated normal, p21 induces high levels of activated JNK. GST also strongly blocks induction of oocyte maturation by protein kinase C (PKC) which is a critical downstream target of oncogenic but not normal ras-p21. Thus, we have established a new function for GST as a potent physiological inhibitor of the ras-JNK-jun pathway. PMID- 10678585 TI - On the role of hydroxyl radical and the effect of tetrandrine on nuclear factor- kappaB activation by phorbol 12-myristate 13-acetate. AB - Nuclear factor kappaB (NF-kappaB) is considered to be an important target for therapeutic intervention because of its role in the regulation of proinflammatory and profibrotic mediators. The present study examined the role of hydroxyl (*OH) radical and the effect of tetrandrine, an alkaloid extracted from the Chinese medicinal herb Stephania tetrandra, on NF-kappaB activation by a tumor promoter, phorbol 12-myristate 13-acetate (PMA) in human lymphoid T cells (ie, Jurkat cells). Exogenous superoxide dismutase (SOD) enhanced the NF-kappaB activation by PMA, while catalase blocked it. Formate, a scavenger of *OH radical, also was inhibitory, as was deferoxamine, a metal chelator. These data suggest an important role of *OH radical in PMA-induced NF-kappaB activation. Incubation of the cells with tetrandrine prior to the stimulation of the cells was found to inhibit PMA-induced NF-kappaB activation. Tetrandrine activity was so potent that 50 microM of tetrandrine was sufficient to inhibit activation of NF-kappaB completely. Electron spin resonance (ESR) spin trapping was used to investigate the antioxidant action of tetrandrine using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trap. Tetrandrine is an antioxidant for both *OH and superoxide (O2-)radicals. The reaction rate constant of tetrandrine with *OH is 1.4 x 10(10) M(-1)sec(-1), which is comparable with several well established antioxidants, such as ascorbate, glutathione, and cysteine. The Fenton reaction (Fe(II) + H2O2- >Fe(III) + *OH + OH-) and xanthine/xanthine oxidase were used as sources of *OH and O2- radicals. The free radical scavenging activity of tetrandrine is responsible for its inhibition of PMA-induced NF-kappaB activation. PMID- 10678586 TI - Cytogenetics as an aid in the diagnosis of lymphomas. AB - Multiple classifications of lymphomas are available. Generally, distinctions are made to identify low, intermediate, and high-risk groups. Histopathologic differentiation is at times difficult. The revised European-American lymphoma classification (REAL) uses histology, clusters of differentiation markers, histochemistry, and cytogenetics for definitive identification. This work reviews the karyotypic and FISH (fluorescent in situ hybridization) findings in some common lymphomas. B-Cell lymphomas, which make up approximately 85-90% of lymphomas, are associated with cytogenetic changes of +12, 13q14, 14q32, 2p11, and 22q13. Translocations help to support the diagnosis of follicular cell lymphoma t(14;18),(q32;q21), mantle cell lymphoma t(11;14)(q13;q32), and Burkitt's lymphoma t(2;8),t(8;14) and t(8;22). T-Cell lymphomas may show changes in 14q11,7p or 7q. Many of the lymphomas are characterized by complex karyotypic changes. Specific FISH probes are useful in determining characteristic or identifying marker chromosomes. Cytogenetic and FISH studies aid in the diagnosis, correct classification, and evaluation of therapy for a variety of lymphomas. PMID- 10678587 TI - Acute renal artery and vein thrombosis after renal transplant, associated with a short partial thromboplastin time and factor V Leiden mutation. AB - Renal graft thrombosis is a rare but devastating complication of renal transplantation. It accounts for one-third to one-half of early graft losses. We report a patient with acute renal artery and vein thrombosis associated with abnormally short activated partial thromboplastin time (aPTT) and factor V Leiden mutation. Vascular thrombosis developed on the ninth post-transplant day and led to a graft loss. Before transplantation, the patient had three episodes of thrombosis of arteriovenous access for hemodialysis. Our case illustrates the importance of investigating pretransplant patients for hypercoagulable states, particularly those with short aPTT. PMID- 10678588 TI - Urine protein electrophoresis and immunofixation electrophoresis supplement one another in characterizing proteinuria. AB - Urine protein electrophoresis (UPE) is often considered to have limited usefulness in evaluating proteinuria that is not associated with gammopathies. Unusual protein bands that are detected by UPE are commonly characterized by immunofixation electrophoresis (IFE). In this paper, electrophoretic gel patterns are shown to illustrate the greater sensitivity of IFE, compared to UPE. However, UPE remains useful for three applications: (1) UPE provides distinctive patterns that can indicate the source of proteinuria and is useful in assessing renal diseases that are independent of gammopathy; (2) combined use of UPE and IFE can avoid misinterpretations and repeated analyses of urine proteins, and (3) UPE can be used in conjunction with IFE to improve the quantitation of Bence-Jones proteinuria (BJP). PMID- 10678589 TI - Modification of screening immunoassays to detect sub-threshold concentrations of cocaine, cannabinoids, and opiates in urine: use for detecting maternal and neonatal drug exposures. AB - Testing for drugs of abuse in urine is commonplace in emergency departments and neonatal units. However, the clinical sensitivity of immunochemical screening methods is limited by the threshold concentrations used to distinguish between positive and negative specimens. Immunochemical screening methods for cocaine metabolite (benzoylecgonine), cannabinoids, and opiates in urine were recalibrated to detect drugs at lower threshold concentrations. The precision and linearity of the signals at the modified thresholds were verified by diluting drug-positive urine specimens to concentrations below the conventional cutoff concentration and measuring the rate signals in triplicate. To assess the clinical performance of the modified methods, specimens that tested negative using the unmodified assays were re-screened at the lower threshold, and specimens that re-screened positive were submitted for gas chromatographic/mass spectrometric (GC/MS) confirmation. Reproducibility of sub-threshold measurements was comparable to the unmodified assays, and rate separations between successive dilutions were sufficient to give semi-quantitative results. Using the lower thresholds, drugs were detected in 4-5% of the subjects that had screened negative at the conventional threshold concentration. GC/MS analysis confirmed the presence of cannabinoids and cocaine metabolite in 74% and 84%, respectively, of urine specimens that re-screened positive. Morphine, codeine, hydromorphone, or hydrocodone was detected by GC/MS analysis in 31% of opiate-positive re screens. PMID- 10678590 TI - Expression of stress proteins HSP 72 & HSP 32 in response to endotoxemia. AB - Pretreatment with heat decreases mortality and acute lung injury in the rat septic shock model, presumably by the production of heat shock proteins (HSP). However, endotoxin, a severe cell stresser, has not been shown to induce HSP 70. We investigated the effects of severe endotoxemia on the expression of specific protective stress proteins, including HSP 72 (inducible HSP 70), HSP 32 (heme oxygenase-1), and HSP 90. Fifteen rats received intravenously either 3 mg/kg of endotoxin (E. coli O127:B8 lipopolysaccharide, LPS) (n=9) or saline (n=6). Two hr later the spleen was removed and splenocytes were separated into three groups and analyzed for specific HSP by Western blot. In Group 1, both endotoxin-treated and saline-treated splenocytes were incubated for 3 hr at 37 degrees C. In Group 2, the splenocytes were washed twice, then heat shocked for 30 min at 42 degrees C and subsequently incubated for 2.5 hr at 37 degrees C. In Group 3, splenocytes were washed twice, then incubated for 3.0 hr at 37 degrees C. HSP 90 & HSP 70c (constitutive) were present in all groups. Consistent with observations by others, HSP 72 was not induced in Group 1. HSP 72 was induced in both the saline treated and endotoxin-treated splenocytes after heating (Group 2). However, in the absence of heat stress, HSP 72 was present in endotoxin-treated but not in saline-treated splenocytes after incubation (Group 3). Conversely, HSP 32, while present in Group 1 splenocytes, was not detected in the endotoxin-treated splenocytes of Group 2 and Group 3, but was present in the saline-treated cells. In conclusion, endotoxemic shock results in induction of HSP 72 and depletion of HSP 32, but only after the cells have been washed and further incubated. PMID- 10678591 TI - Effect of gliotoxin on development of diabetes mellitus in diabetes-prone BB/Wor rats. AB - The object of this investigation was to determine if gliotoxin, an immunomodulating fungal secondary metabolite, is capable of preventing the development of autoimmune diabetes mellitus in diabetes-prone BB/Wor rats. Chronic treatment, consisting of 1 microg gliotoxin/g of body wt administered three times weekly from the age of 30 days through 120 days, reduced the incidence of diabetes from 90% diabetic by 120 days among vehicle-treated animals to 56% diabetic among gliotoxin-treated animals. This result was significant by life table analysis. Animals treated with gliotoxin maintained lower serum glucose levels even in the pre-diabetic state than control (vehicle-treated) rats. Gliotoxin at levels used in this study showed no appreciable effect on the viability of rat insulinoma (RIN 38) cells in culture and only slightly decreased their insulin secretion. Animals chronically treated with gliotoxin showed weight gains comparable to those seen in controls, and the effect of gliotoxin on peripheral blood leukocyte counts was not significant. The possibility that gliotoxin exerted its effect through immunomodulating effects was implied by the loss of white pulp in splenic follicles of gliotoxin-treated animals. PMID- 10678592 TI - Pathoaetiology, epidemiology and diagnosis of hypertension. AB - Hypertension is currently defined in terms of levels of blood pressure associated with increased cardiovascular risk. A cut-off of 140/90 mm Hg is accepted as a threshold level above which treatment should at least be considered. This would give a prevalence of hypertension of about 20% of the adult population in most developed countries. Hypertension is associated with increased risk of stroke, myocardial infarction, atrial fibrillation, heart failure, peripheral vascular disease and renal impairment. Hypertension results from the complex interaction of genetic factors and environmental influences. Many of the genetic factors remain to be discovered, but environmental influences such as salt intake, diet and alcohol form the basis of nonpharmacological methods of blood pressure reduction. Investigation of the individual hypertensive patient aims to identify possible secondary causes of hypertension and also to assess the individual's overall cardiovascular risk, which determines the need for prompt and aggressive therapy. Cardiovascular risk can be determined from (i) target organ damage to the eyes, heart and kidneys; (ii) other medical conditions associated with increased risk; and (iii) lifestyle factors such as obesity and smoking. Secondary causes of hypertension are individually rare. Screening tests should be initially simple, with more expensive and invasive tests reserved for those in whom a secondary cause is suspected or who have atypical features to their presentation. The main determinants of blood pressure are cardiac output and peripheral resistance. The typical haemodynamic finding in patients with established hypertension is of normal cardiac output and increased peripheral resistance. Treatment of hypertension should initially use nonpharmacological methods. Selection of initial drug therapy should be based upon the strength of evidence for reduction of cardiovascular mortality in controlled clinical trials, and should also take into account coexisting medical conditions that favour or limit the usefulness of any given drug. Given this approach, it would be reasonable to use a thiazide diuretic and/or a beta-blocker as first-line therapy unless there are indications to the contrary. Individual response to given drug classes is highly variable and is related to the underlying variability in the abnormal pathophysiology. There are data to suggest that the renin-angiotensin system is more important in young patients. The targeting of this system in patients under the age of 50 years with a beta-blocker (or ACE inhibitor), and the use of a thiazide diuretic (or calcium antagonist) in patients over 50 years, may enable blood pressure to be controlled more quickly. PMID- 10678593 TI - Pharmacoeconomic considerations in the management of hypertension. AB - Hypertension is highly prevalent in developed and developing countries (more than 30% of the adult population when a threshold value of 140/90 mm Hg is selected). It constitutes one of the major cardiovascular risk factors and accounts for more than 5% of total deaths worldwide. The economic impact of hypertension is enormous, representing $US23.74 billion in the US in 1995 and approximately $US1685 million in Spain in 1994. Direct costs amount to more than 50% of the total costs of hypertension, and almost 70% of these are attributable to drug treatment. Furthermore, hypertensive patients use medical services 50% more than normotensive individuals, and hypertension represents one of the 3 leading causes of visits to primary healthcare centres. When considering the cost effectiveness of hypertension treatment, there is no doubt that it is cost effective in comparison with other interventions, although some controversies exist, mainly with respect to mild-to-moderate hypertension and to the long term versus short term benefits. The controversy about the absolute risk of hypertension influences the cost-effectiveness analysis. Because of the limitations of the available cost effectiveness analyses, it is currently impossible to recommend the use of any particular antihypertensive drug for all patients with hypertension. Consequently, the choice of antihypertensive in any patient should be guided by clinical experience and the recommendations of the present international guidelines. PMID- 10678594 TI - Diuretics as a basis of antihypertensive therapy. An overview. AB - Diuretics have been, except for during a few recent years, the most commonly used therapy for hypertension. Although use of these agents fell significantly in the early 1990s, since then it has begun to increase again. Their recent return to popularity reflects 3 major factors: (i) recognition of the effectiveness of much lower dosages than previously used, thereby providing good antihypertensive activity with fewer adverse effects; (ii) the excellent reductions in morbidity and mortality achieved by low dosage diuretic-based therapy in multiple randomised controlled trials in elderly patients with hypertension; and (iii) the increasing recognition that some diuretic-induced shrinkage of effective blood volume is essential for the adequate treatment of many, if not most, patients with hypertension. Therefore, diuretics will probably continue to be the basis for antihypertensive therapy, and the indapamide sustained release 1.5 mg formulation provides all the essential characteristics of diuretic therapy. PMID- 10678595 TI - Clinical positioning of indapamide sustained release 1.5mg in management protocols for hypertension. AB - Indapamide sustained release (SR) 1.5mg is a new galenic formulation that is characterised by a relatively constant plasma concentration at steady state, with only minor fluctuations during the 24-hour period. A dose-titration study of 3 doses of indapamide SR (1.5, 2 and 2.5mg) given once daily has shown that the 3 dosages are equipotent in lowering blood pressure, and have an effect similar to that of indapamide immediate-release (IR) 2.5mg; all were statistically more effective than placebo. The percentage of hypertensive patients whose serum potassium was less than 3.4 mmol/L was significantly lower after indapamide SR 1.5mg than after indapamide IR 2.5mg. Neither indapamide formulation had any significant effects on lipid profile, glucose, urea and serum creatinine; only uric acid was slightly raised during the 2-month study. In an equivalence study, indapamide SR 1.5mg and IR 2.5mg produced similar blood pressure reductions (within the equivalence limit of +/-5mm Hg), whereas the percentage of patients whose serum potassium fell to less than 3.4 mmol/L was lower in the IR 1.5mg group than in the SR 2.5mg group. Antihypertensive treatment with indapamide SR 1.5mg once daily produced reductions in blood pressure in elderly patients with systolic/diastolic or isolated systolic hypertension that were similar to reductions with amlodipine 5 mg/day. The incidence of adverse effects was very low in all studies with indapamide SR 1.5mg and very similar to that in the placebo group, confirming thereby the improvement in the efficacy: tolerance ratio with the new indapamide compound. PMID- 10678596 TI - Paralytic vs. "nonparalytic" polio: distinction without a difference? AB - Nonparalytic polio (NPP) is commonly thought to be synonymous with "abortive polio," in which the poliovirus neither entered the central nervous system nor damaged neurons. Described are two epidemic illness-"The Summer Grippe" and Iceland disease-apparently caused by a low virulence but neuropathic type 2 poliovirus. Studies show that neuronal lesions in the brain and spinal cord and muscle weakness were common in NPP, and epidemiologic studies document late-onset weakness and fatigue in 14% to 42% of NPP survivors. These findings indicate that clinicians should not require a history of paralytic polio, electromyographic evidence of denervation, and new muscle weakness for the diagnosis of "Postpolio Syndrome" but should be aware that NPP, and possibly even poliovirus-induced "minor illnesses," can be associated with acute central nervous system damage and late-onset muscle weakness and fatigue. PMID- 10678597 TI - Nonparalytic polio and postpolio syndrome. AB - We describe four cases of postpolio syndrome with typical histories, physical examination results, and electrodiagnostic evidence of extensive anterior horn cell disease, as well as the putative pathophysiology of postpolio syndrome in persons with histories of nonparalytic polio and the diagnostic implications for individuals older than 40 yr of age who are experiencing unexplained new weakness, fatigue, and muscle or joint pain. Although the diagnosis of postpolio syndrome traditionally has required a remote history of paralytic polio, many persons such as the ones described here with typical symptoms of postpolio syndrome have no clear history of paralytic disease and are being misdiagnosed. With this in mind, we believe that the diagnostic criteria for postpolio syndrome should be modified to include the following: a history of remote paralytic polio or findings on history, physical examination results, and laboratory studies compatible with poliovirus damage of the central nervous system earlier in life. PMID- 10678598 TI - Late functional loss in nonparalytic polio. PMID- 10678599 TI - Kyphoscoliosis ventilatory insufficiency: noninvasive management outcomes. AB - OBJECTIVE: To determine the effects on symptoms, pulmonary function, sleep, and other clinical variables of treating kyphoscoliosis-associated chronic alveolar hypoventilation with nocturnal nasal ventilation. DESIGN: Sixteen patients with kyphoscoliosis were treated with nocturnal nasal ventilation delivered by volume cycled (seven patients) and pressure-cycled (nine patients) ventilators. Dyspnea, morning headaches, fatigue, hypersomnolence, and perceived sleep quality were assessed. RESULTS: All pretreatment symptoms improved significantly with nasal ventilation. Likewise, PaO2 (mm Hg), PaO2/FlO2, PaCO2 (mm Hg), pH, and forced vital capacity (in milliliters and as a percentage of predicted normal) significantly improved with treatment. Maximum inspiratory pressures and maximum expiratory pressures also significantly increased. Tidal volumes increased significantly and breathing frequency decreased (not significant). Although perceived sleep quality improved, as well as sleep oxyhemoglobin saturation, there was no significant change in sleep architecture. Hospitalization days for respiratory difficulties also decreased from 10.9 +/- 13.3 days in the 6 mo before intermittent positive-pressure ventilation to 0 days during the first 6 mo of treatment. CONCLUSIONS: Although not apparently affecting sleep architecture, nocturnal nasal ventilation can significantly improve nocturnal and daytime blood gases, pulmonary function, and symptoms of hypoventilation for patients with severe kyphoscoliosis. PMID- 10678600 TI - Balance, mobility, and falls among elderly African American women. AB - OBJECTIVE: To compare balance, mobility, recent falls, and injuries among elderly African American and white women. DESIGN: This was a nonexperimental study. Participants, who were older than 65 yr of age, able to walk at least 30 ft, not residing in a nursing home, and with no acute medical problems, were recruited from 17 senior citizens' community centers. RESULTS: Compared with white women (n=180), African American women (n = 118) took fewer medications, had greater body mass indexes, had less muscle strength, and had more medical conditions and neurologic abnormalities. Additionally, these women were less active and had poorer performances on an obstacle course. The two groups had a similar histories of falls and injuries. For both groups, activity level and neurologic findings were predictors of obstacle course performance. For white women, muscle strength was an additional predictor of obstacle course performance. An additional predictor for African American women was range of motion. CONCLUSION: The poorer balance and mobility of African American women compared with white women may have consequences such as their functional dependence, resulting in their greater use of hospitals and formal and informal health services. PMID- 10678601 TI - Temporal effects of isometric contraction maneuvers on threshold sural amplitude. AB - OBJECTIVE: To investigate the effect of isometric biceps brachii contraction and neck flexion on the time course of threshold sural amplitude. DESIGN: Twelve healthy subjects, who were asked to lie supine on an examination bench, performed 1 min of muscle contraction. The sural sensory nerve action potential was recorded before, immediately after, and at 2-min intervals after muscle contraction. The preexercise level of stimulus intensity remained unchanged for sural readings throughout the entire course of the experiment. RESULTS: The temporal changes in sensory nerve action potential amplitudes for both maneuvers were similar (P = 0.9734, two-way interaction). The mean sural amplitude after neck flexion increased from 6.0 +/- 2.9 microV (SD) to 10.6 +/- 6.6 microV (SD) 10 min after contraction. Similarly, mean sural amplitude increased from 6.5 +/- 1.8 microV (SD) to 14.5 +/- 9.7 microV (SD) 8 min after biceps brachii contraction. Statistical analysis performed using repeated measures with post hoc least significant difference showed a significant temporal effect in the two groups (P = 0.04). CONCLUSION: The temporal responses of threshold sural amplitudes after isometric biceps brachii contraction and central reinforcement neck flexion maneuvers are nearly identical with regard to increase in the amplitude. PMID- 10678602 TI - Botulinum toxin injection of spastic finger flexors in hemiplegic patients. AB - OBJECTIVE: To assess the outcomes of botulinum toxin injection of spastic finger flexors followed by intensive training of finger extensors. DESIGN: Fourteen subjects with chronic hemiplegia spasticity of the upper limb had electromyographic-guided botulinum toxin injection into the long finger flexors. All patients presented with minimal active finger extension with the wrist flexed, sustained clonus of the finger flexors, functional proximal arm function, and absence of fixed contracture. Cadaver dissections directed selection of two injection sites: the flexor digitorum sublimis and the flexor digitorum profundus. Fifty mouse units of botulinum toxin were injected into each muscle. After injection, the subjects were instructed in a home program of stretching the long finger flexors, upper limb weight bearing with a weight-bearing splint, and exercise to improve finger extension control. RESULTS: Compared with preinjection measures, assessment the first week after the initial injection showed significantly reduced tone, reduced clonus, and greater active finger extension with the wrist in the neutral position. Four months later, the Ashworth scale increased to preinjection levels in the six subjects with repeated injections but was again decreased postinjection. Active finger extension with the wrist in the neutral position and clonus showed a statistically nonsignificant trend toward cumulative improvement after the second injection. CONCLUSION: The greatest change in finger extension and spasticity reduction occurred after the first injection. Continued significant improvement in finger extension was not observed. PMID- 10678603 TI - Treatment of myofascial pain. AB - OBJECTIVE: To investigate the effectiveness of ultrasound treatment and trigger point injections in combination with neck-stretching exercises on myofascial trigger points of the upper trapezius muscle. DESIGN: Depression and anxiety associated with chronic pain were assessed using the Beck Depression Inventory (BDI) and the Taylor Manifest Anxiety Scale (TMAS). The study population comprised 102 patients who had myofascial trigger points in one side of the upper trapezius. The patients were randomly assigned to one of three groups: group 1 received ultrasound therapy to trigger points in conjunction with neck-stretching exercises; group 2 received trigger point injections and performed neck stretching exercises; and group 3, the control group, performed neck-stretching exercises only. Treatment effectiveness was assessed using subjective pain intensity (PI) with a visual analog scale, pressure pain threshold (PT) with algometry, and range of motion (with a goniometer) of the upper trapezius muscle. RESULTS: Compared with the control group, patients in groups 1 and 2 had a statistically significant reduction in PI, an increase in PT, and an increase in range of motion. There were no statistically significant differences between treatment groups 1 and 2. Although not statistically significant, patients in the control group had better results at the 3-mo follow-up. The BDI scores indicated depression in 22.9% of the patient, with 4.8% of the patients having severe depression. High anxiety scores on the TMAS were present in 89.3% of the patients. When BDI and TMAS scores were compared with PI or PT levels, no significant correlations were found, but when compared with pain duration before treatment, correlations were significant. CONCLUSIONS: Patients with myofascial pain syndrome had higher scores for anxiety than for depression. When combined with neck stretching exercises, ultrasound treatment and trigger point injections were found to be equally effective. PMID- 10678604 TI - Effects of botulinum toxin A on upper limb spasticity in children with cerebral palsy. AB - OBJECTIVE: Botulinum toxin A inhibits presynaptic release of acetylcholine at the neuromuscular junction and has reportedly been successful in the treatment of spastic disorders. This prospective study attempted to determine whether botulinum toxin A injection resulted in clinically measurable gains for 4 mo. DESIGN: Measurements were obtained from 32 children (range, 1-18 yr; average age, 6.9 yr) with hemiplegic or quadriplegic cerebral palsy before and at 1, 3, and 4 mo after botulinum toxin A injections. Spasticity was measured using the Modified Ashworth Scale for 12 different joints. RESULTS: Results showed that spasticity as measured by Ashworth scores for elbow and wrist extension clearly declined (P < 0.02) by 1 mo after botulinum toxin A injection, and diminished spasticity continued for 3-4 mo. Caregivers reported improvement in subjectively rated management, appearance, and function. However, patient response to botulinum toxin A injection was not predictable. Age had no significant relationship to gains. CONCLUSIONS: Further research is needed on the use of botulinum toxin A to diminish spasticity and improve function. PMID- 10678606 TI - Dosing practices of physicians for anticoagulation with warfarin during inpatient rehabilitation. AB - OBJECTIVE: To determine the percentage of international normalized ratios (INRs) maintained within the therapeutic range for patients receiving chronic anticoagulation treatment with warfarin during inpatient rehabilitation. DESIGN: A consecutive, 4-month, retrospective chart review of all patients receiving oral anticoagulation treatment was conducted in a large academic rehabilitation center. The percentage of INRs within and out of the therapeutic range, frequency of blood samples, length of therapy, and warfarin dose prescribed by physicians were calculated. A total of 181 patients receiving chronic anticoagulation treatment were identified. A total of 3,709 blood samples were analyzed. In 74 patients, the primary physician recommended a therapeutic range (Group 1). In the remaining 107 patients, no therapeutic range was specified, and a target INR range of 2.0-3.0 was assumed (Group 2). RESULTS: In Group 1, the INRs were in the recommended range in 38.2% of all blood samples. In Group 2, 37.6% of all blood drawn was within an INR range of 2.0-3.0. Statistical analysis showed that no better accuracy was obtained when the INR range was predefined by a physician (Group 1) or assumed to be in the 2.0-3.0 range (Group 2; P = 0.839). CONCLUSIONS: Despite frequent physician monitoring, this study demonstrates the difficulty in maintaining INRs within therapeutic ranges for patients receiving oral anticoagulation. An overall tendency for underdosing is observed. Improvement is necessary, given the high morbidity and mortality associated with insufficient anticoagulation in rehabilitation inpatients. PMID- 10678605 TI - Effect of history and exam in predicting electrodiagnostic outcome among patients with suspected lumbosacral radiculopathy. AB - OBJECTIVE: To determine the extent to which the history and physical examination predict the outcome of the electrodiagnostic (EDX) evaluation in patients with suspected lumbosacral radiculopathy. DESIGN: Data for 170 subjects referred for low-back and lower limb symptoms were prospectively collected at five EDX laboratories. The sensitivity, specificity, positive and negative predictive values, and odds ratios were determined for symptoms and neurologic signs. RESULTS: Symptoms were not significantly associated with an EDX study or a lumbosacral radiculopathy. The physical examination was better at predicting that an EDX study would be abnormal in general than it was at predicting a lumbosacral radiculopathy in particular. Of those subjects with normal physical examinations, 15%-18% still had abnormal EDX findings. CONCLUSIONS: In a population of patients referred for an EDX study, the history and physical examination alone cannot reliably predict electrodiagnostic outcome. PMID- 10678607 TI - The future of physical medicine and rehabilitation. PMID- 10678608 TI - The Uniform Data System for Medical Rehabilitation: report of first admissions for 1998. PMID- 10678609 TI - Heparin-induced hyperkalemia confirmed by drug rechallenge. AB - Subcutaneous heparin is commonly used as a prophylaxis against deep venous thrombosis in a wide variety of hospitalized patients. As with most medications, heparin has a significant side effect profile; heparin-induced hyperkalemia is an unusual but well described side effect. To increase awareness of heparin-induced hyperkalemia and of those patients at greatest risk, we present two cases of documented hyperkalemia induced by heparin and reconfirmed by drug rechallenge. PMID- 10678610 TI - Improving the workers compensation system: case management perspective. PMID- 10678611 TI - Rough sets: a knowledge discovery technique for multifactorial medical outcomes. AB - Rough sets is a fairly new and promising technique for data mining and knowledge discovery from databases. This tutorial article presents the fundamentals of rough set theory in a nontechnical manner and outlines how the technique can be used to extract minimal if-then rules from tables of empirical data that either fully or approximately describe given example classifications. An example application for prediction of ambulation for patients with spinal cord injury is given. Because such rules are readily interpretable, they can be inspected to yield possible new insight into how various contributing factors interact and, thus, serve as hypothesis generators for further research. Additionally, the set of mined rules may function as a classifier of new, unseen cases. PMID- 10678612 TI - Starting inhaled corticosteroids in asthma: when, how high, and how long. PMID- 10678613 TI - The lung in inflammatory bowel disease. PMID- 10678614 TI - Low- and high-dose fluticasone propionate in asthma; effects during and after treatment. AB - The dose dependency of the effects of inhaled corticosteroids on markers of asthmatic airway inflammation have not been well studied. There is a need to study the dose/response effects on this inflammation. In order to determine the dose/response effects of fluticasone propionate (FP), 24 asthmatic subjects were randomized to low- (100 microg x day(-1)) or high-dose (1,000 microg x day(-1)) FP for six weeks followed by placebo for 3 weeks. During treatment, the median increase in forced expiratory volume in one second (FEV1)was 12% in the high-dose group (p<0.05) and 10% in the low-dose group (p<0.05) (p>0.05 between groups); the median decrease in the percentage of sputum eosinophils was 93% in the high dose group (p<0.05) and 46% in the low-dose group (p<0.05) (p>0.05 between groups). Symptoms, salbutamol use, morning peak flow, provocative concentration of methacholine causing a 20% fall in FEV1 (PC20), sputum eosinophil cationic protein concentration and tryptase activity improved significantly in both groups (p<0.05), but only the improvement in salbutamol use was greater in the high-dose group (p<0.05). During the run-out, the improvements in FEV1 and PC20 were rapidly reversed in both groups, but the improvements in peak flow and tryptase activity persisted; the improvement in sputum eosinophil concentration persisted only in the high-dose group (p<0.05). It was concluded that dose/response effects for FP are not easily demonstrable because low-dose FP is quite effective. For most outcomes, the effects of high- and low-dose FP are relatively short-lived after treatment is stopped. This finding raises questions about the extent to which inhaled corticosteroids are disease-modifying in asthma. PMID- 10678615 TI - Effect of inhaled fluticasone propionate on airway responsiveness in treatment naive individuals--a lesser benefit in females. AB - A randomized double-blind placebo-controlled parallel group study with inhaled fluticasone propionate over 6 weeks, designed to quantify the beneficial effect on airway responsiveness, and so assess whether short pulses of intermittent prophylactic treatment might serve as an alternative means of managing mild asthma, is reported. The 20-50-yr-old participants, who were recruited from an epidemiological study of the general population, had never knowingly received any regular treatment for asthma. Fluticasone propionate at the maximum recommended dose level (2,000 microg daily) and placebo were administered via metered-dose inhalers, and airway responsiveness was quantified conventionally by the provocative dose of methacholine causing a 20% fall in forced expiratory volume in one second (FEV1) (PD20) at 2-week intervals during the treatment phase and at various intervals subsequently. Compared with placebo fluticasone propionate was associated with a highly significant decrease in airway responsiveness (1.9 doublings of the geometric mean PD20), which was maximal at the end of the 6-week treatment period. No persisting benefit was detectable at the next measurement 2 weeks later, or thereafter. Multiple linear regression analysis showed that the magnitude of the fluticasone propionate effect was significantly greater in males than in females (3.2 versus 1.2 doublings respectively of the geometric mean PD20), but was uninfluenced by current smoking, age or FEV1. In conclusion, in the absence of any possibility of tachyphylaxis, inhaled fluticasone propionate at this dose causes a steadily increasing improvement in airway responsiveness over a 6-week period, which is modified by sex but lost almost immediately on treatment cessation. Short pulses of intermittent prophylactic treatment would not, therefore, be useful as a means of managing mild asthma. PMID- 10678616 TI - Asthma management in five European countries: doctors' knowledge, attitudes and prescribing behaviour. Drug Education Project (DEP) group. AB - The aim of the study was to examine the relationship between guideline recommendations on asthma management, and the performance of doctors in five different European health care contexts. Knowledge, attitudes and prescribing behaviour of doctors recruited to an educational project was investigated. A total of 698 general practitioners from Germany, The Netherlands, Norway and Sweden, and 94 specialists from the Slovak Republic participated. A questionnaire was used to assess their knowledge and attitudes. Antiasthmatic drugs dispensed to their patients reflected their prescribing behaviour. In response to questions on how to treat chronic asthma, most doctors were in agreement with guideline recommendations. In practice, however, the proportion of asthma patients receiving inhaled steroids varied almost twofold, ranging 31% in Germany to 58% in The Netherlands. On questions related to exacerbation of asthma, German and Slovakian doctors often preferred treatment with antibiotics to steroids. They also more often associated yellow-green sputum with bacterial infection. In conclusion, although many doctors in different health care contexts have accepted the recommendations given in guidelines, the proportion of their patients treated accordingly differed. German and Slovakian doctors seem to attach less importance to the inflammatory features of asthma than the doctors from the other three European countries. PMID- 10678618 TI - Neutrophils in induced sputum arise from central airways. AB - A high neutrophil count is often found in induced sputum compared to bronchoalveolar lavage fluid (BALF). This study investigated whether such a high neutrophil count may be a response to inhaled hypertonic saline or that the airway compartment sampled by induced sputum has a higher concentration of neutrophils than BALF. Saliva and induced sputum samples were taken at 10 and 20 min following inhalation of 3% hypertonic saline from an ultrasonic nebulizer in 12 healthy nonsmoking subjects. Four days later the 12 subjects underwent bronchoscopy (six following inhalation with 3% saline). Tracheal, proximal bronchial secretions, bronchial washings and BALF samples were obtained. The neutrophil count (% total leukocytes) increased significantly in saliva at 10 and 20 min post nebulization with 3% saline, although there was no change in neutrophils in induced sputum at 10 and 20 min. There was no significant difference in neutrophil count in the subjects who inhaled 3% saline as compared to those who did not, in secretions from the trachea, proximal bronchi, bronchial washings and BALF. Neutrophil counts were significantly higher in the trachea, proximal bronchi and bronchial washings as compared to BALF (p<0.001). It is concluded that neutrophil counts in healthy subjects increase from the peripheral towards the proximal airways, in the absence of hypertonic saline-induced changes. This suggests that the relatively high neutrophil count in induced sputum arises from the proximal airways and is not a response to inhaled hypertonic saline during the procedure. PMID- 10678617 TI - Bronchial hyperresponsiveness and airway inflammation markers in nonasthmatics with allergic rhinitis. AB - Bronchial hyperresponsiveness (BHR) is a characteristic feature of asthma which is often associated with airways inflammation. However, some patients with allergic rhinitis and no clinical evidence of asthma also exhibit BHR. This study therefore investigated whether inflammatory cell infiltrate is present in the induced sputum of nonasthmatic subjects with allergic rhinitis during the pollen season and examined its relationship with airway hyperresponsiveness to inhaled methacholine and adenosine 5'-monophosphate (AMP). Twenty subjects (12 allergic rhinitis, eight nonallergic controls) underwent methacholine and AMP challenge and sputum induction with hypertonic saline on separate days. Cell differentials were calculated from whole sputum samples. A significantly greater number of eosinophils was found in the sputum of nonasthmatic subjects with allergic rhinitis compared to that of nonallergic controls, their median (range) percentages being 17.5 (4-47) and 1.5 (0-5) (p<0.001) respectively. Although sputum eosinophilia failed to be significantly associated with methacholine responsiveness (r(s)=-0.50; p=0.095), the provocative concentration of AMP causing a 20% fall in forced expiratory volume in one second correlated strongly and significantly with the absolute number of eosinophils (r(s)= -0.73; p=0.007). Eosinophil cationic protein levels in the sputum of rhinitic subjects were significantly elevated compared to controls and correlated with eosinophil number (r(s)=0.67; p=0.017). These findings support the view that bronchial eosinophilia alone is insufficient to cause asthmatic symptoms. Diverse agonists for assessing bronchial hyperresponsiveness are selectively associated with airway inflammation in allergic rhinitis. PMID- 10678619 TI - Clinical and radiological characteristics of lung disease in inflammatory bowel disease. AB - The pulmonary associations of inflammatory bowel disease (IBD) are poorly characterized. The clinical, physiological and high-resolution computed tomographic thorax characteristics of the lung disease in patients with IBD presenting with respiratory symptoms are described. Detailed clinical information was obtained and standard pulmonary physiological tests and thorax high resolution computed tomography performed on 14 patients with ulcerative colitis (UC) and three with Crohn's disease (CD), 10 male, aged 38-83 yrs. Respiratory symptoms had been present for 2-50 yrs and extraintestinal manifestations were present in three (17.6%). Normal pulmonary physiology (six patients) was associated with the high resolution computed tomographic changes of bronchiectasis, mosaic perfusion and air trapping suggestive of obliterative bronchiolitis and a pattern of centrilobular nodules and branching linear opacities ("tree in bud" appearance) suggestive of either cellular bronchiolitis or bronchiolectasis with mucoid secretions. Bronchiectasis was found in 13 patients (11 UC, 2 CD), 11 had air trapping and five had a "tree in bud" appearance on computed tomography. One patient had a predominantly peripheral reticular pattern at the lung bases similar to that found in cryptogenic fibrosing alveolitis and one patient had a mixed reticular and ground-glass pattern in the midzones with a patchy distribution in the central and peripheral portions of the lungs with air trapping. Eleven patients (three with alveolitis) exhibited a clinical and/or physiological response to steroids. Pulmonary abnormalities in ulcerative colitis and Crohn's disease can present years after the onset of the bowel disease and can affect any part of the lungs. Early recognition is important as they can be strikingly steroid-responsive. PMID- 10678620 TI - B7-1, B7-2 and class II MHC molecules in idiopathic pulmonary fibrosis and bronchiolitis obliterans-organizing pneumonia. AB - Interstitial lung diseases are thought to be associated with the infiltration of activated T-lymphocytes. To induce an effective immune response, antigen presenting cells have to not only present antigenic peptide with major histocompatibility complex (MHC) molecules to T-lymphocytes but also express B7 molecules. Therefore, the expression of B7-1, B7-2 and class II MHC molecules was investigated in lung tissues from patients with idiopathic pulmonary fibrosis (IPF) and bronchiolitis obliterans-organizing pneumonia (BOOP), and in normal lung parenchyma as a control, using immunohistochemical localization. B7-1 and B7 2 were aberrantly expressed in bronchiolar and alveolar epithelial cells, and class II MHC molecules were also aberrantly expressed in bronchiolar epithelial cells in IPF. B7-1 was aberrantly expressed in bronchiolar epithelial cells in BOOP. There was no significant difference in the expression of these proteins in alveolar macrophages between IPF and control subjects. However, B7-2 and class II MHC molecule expression in alveolar macrophages was decreased in BOOP compared with that in control subjects. Expression of CD28 and CTLA4, receptors for B7 molecules, was detected in infiltrating lymphocytes in lung tissues in IPF and BOOP. It was concluded that bronchiolar and alveolar epithelial cells may actively participate in the pathophysiology of idiopathic pulmonary fibrosis through the aberrant expression of B7 and class II major histocompatibility complex molecules. The dysregulation of these molecules in epithelial cells may lead to the activation of autoreactive T-lymphocytes, which might contribute to the pathogenesis of fibrosing lung diseases. PMID- 10678621 TI - Pulmonary function and complications following chemotherapy and stem cell support in breast cancer. AB - Pulmonary complications are frequent in patients treated with high-dose chemotherapy and autologous bone marrow transplantation for breast cancer or other solid tumours. This study analyses the development of lung toxicity, changes in respiratory function and occurrence of clinical symptoms in a group of 24 patients (mean age 46+/-7 yrs) who underwent high-dose sequential chemotherapy (HDS) with autologous peripheral blood stem cell (PBSC) support for high risk breast cancer. Clinical examination, chest radiography and lung function tests were performed before the HDS and 1 and 3 months following transplantation. Only one patient developed acute interstitial pulmonary disease which resolved after prednisone therapy. No patients developed infectious complications after transplantation. Baseline respiratory function was normal for most of the parameters. Only lung diffusing capacity of the lung for carbon monoxide (TL,CO) and maximal inspiratory pressure were below the normal range. Following PBSC transplantation only one patient had an altered vital capacity while 72.3% of patients had reduced TL,CO values at 1 month and 54.5% at 3 months after transplantation. Maximal expiratory flow at 25% forced vital capacity, TL,CO and maximal expiratory pres-sure were significantly reduced after 1 month but recovered slightly by 3 months. Arterial oxygen tension between baseline and both follow-up evaluations declined significantly in patients seropositive for human cytomegalovirus. It is concluded that this high-dose sequential chemotherapy regimen is acceptably safe since no pulmonary related mortality or respiratory infectious complications were observed. The only lung function alteration induced was an isolated diffusing capacity of the lung for carbon monoxide impairment, clinically negligible and partially recovered within 3 months. PMID- 10678622 TI - Macrolide antibiotics inhibit nitric oxide generation by rat pulmonary alveolar macrophages. AB - There is evidence that macrolide antibiotics are effective in the treatment of chronic airway inflammatory diseases, probably through actions other than their antibacterial properties. In order to determine whether macrolides affect the nitric oxide-generating system in the respiratory tract, rat pulmonary alveolar macrophages (PAMs) were studied in vitro. The release of NO was assessed by direct measurement with a specific amperometric sensor for this molecule, and the expression of type II NO synthase (NOS) messenger ribonucleic acid (mRNA) was determined by Northern blotting. Incubation of PAMs with lipopolysaccharide from Escherichia coli and recombinant human interferon-gamma caused release of NO, which was accompanied by induction of type II NOS mRNA. The release of NO was reduced by coincubation of cells with the macrolides erythromycin, clarithromycin and josamycin in a concentration-dependent manner, the maximal inhibition being 73+/-10, 81+/-6 and 84+/-9%, respectively, but was not altered by amoxycillin or cefaclor. These macrolides likewise inhibited the induction of type II NOS mRNA, whereas no inhibitory effects were observed with amoxycillin or cefaclor. These results suggest that macrolide antibiotics specifically inhibit type II NO synthase gene expression and consequently reduce NO production by rat pulmonary alveolar macrophages, which might result in attenuation of airway inflammation. PMID- 10678623 TI - Budesonide and formoterol inhibit ICAM-1 and VCAM-1 expression of human lung fibroblasts. AB - The glucocorticoid budesonide and the long-acting beta2-adrenoceptor agonist formoterol are used in asthma therapy for their anti-inflammatory and bronchodilating effects, respectively. Since expression of adhesion molecules on resident cells in the lung plays an important role in asthmatic inflammatory responses, the effects of these drugs on the cytokine-induced intercellular adhesion molecule-1 (ICAM)-1 and vascular cell adhesion molecule-1 (VCAM)-1 expression of human lung fibroblasts were investigated. Budesonide and formoterol were added in the absence or presence of interleukin (IL)-1beta, tumour necrosis factor-alpha (TNF-alpha), interferon gamma (IFN-gamma) or IL-4 to human lung fibroblasts; ICAM-1 and VCAM-1 expression were measured after 8 h using a cell surface enzyme linked immunosorbent assay (ELISA). It was found that both budesonide and formoterol significantly inhibited (p<0.05) the increased expression of ICAM-1 and VCAM-1 after stimulation with IL-1beta (maximal inhibition (median (25-75% percentiles) 50 (48-52) and 61% (42-69), respectively, with budesonide and 55 (50-73) and 86% (64-94), respectively, with formoterol (10(-7) M)), TNF-alpha (maximal inhibition 49 (46-57) and 57% (44-68), respectively, with budesonide and 44 (40-75) and 62% (52-83) respectively, with formoterol), IFN-gamma (maximal inhibition 64% (41-67) with budesonide and 39% (29-49) with formoterol for ICAM-1) and IL-4 (maximal inhibition 82% (69-92) with budesonide and 43% (33-67) with formoterol for VCAM-1) in a dose-dependent manner. The results show that budesonide, as well as formoterol, in probably clinically relevant concentrations inhibits cytokine-induced adhesion molecule expression on human lung fibroblasts from a concentration of 10(-9) M. This inhibitory effect on resident cells may have implications for the infiltration of inflammatory cells into pulmonary tissue during therapy with these drugs in asthma. PMID- 10678624 TI - Leukotoxin and its diol induce neutrophil chemotaxis through signal transduction different from that of fMLP. AB - When injected into animals, leukotoxin (Lx) causes acute lung injury which is associated with neutrophils infiltrating the lung tissues. However, the effect of Lx on neutrophils is still unknown, and recently it has been reported that Lx diol, a hydrolyzed metabolite, should be more potent than Lx in vitro. In this study, the authors examined the effect of Lx and its diol on human neutrophils by assessing their chemotactic response, expression of adhesion molecules, and production of peroxides. Both Lx and its diol induced chemotaxis in human neutrophils via an involvement of pertussis toxin-sensitive G-proteins, but they did not influence the expression of adhesion molecules or the production of peroxides. Furthermore, Lx synergistically affected chemotaxis with N-formyl methionyl-leucyl-phenylalanine (fMLP), but not with endothelin-1. Neutrophil chemotaxis induced by both Lx and its diol was inhibited by phosphatidylinositol 3-kinase (PI3-K) inhibitors, but not by protein tyrosine kinase (PTK) inhibitors or by protein kinase C (PKC) inhibitors, whereas fMLP-induced chemotaxis was inhibited by PTK inhibitors, but not by PI3-K inhibitors or by PKC inhibitors. These results suggest that neutrophil chemotaxis induced by both Lx and its diol involves pathways different from those induced by fMLP. In conclusion, both leukotoxin and its diol metabolite induce chemotaxis in human neutrophils in a unique way and may act as important bioactive lipids when considering the pathological mechanism of acute lung injury. PMID- 10678625 TI - Effects of aging on surfactant forms in rats. AB - Surfactant present in the alveolar space exists in two major forms: functional large aggregate forms (LA) and nonfunctional small aggregate forms (SA), but there is no information about the changes of surfactant forms and the rate of conversion of LA to SA in the aged lungs. The purpose of the present study was to investigate the developmental aspects of surfactant forms in newborn, young, middle-aged and aged rats, LA and SA were recovered from alveolar lavages of rats. The rate of conversion from LA to SA was then analysed using a surface-area cycling technique. Age-related changes of saturated phosphatidylcholine (Sat-PC) and surfactant protein A (SP-A) pool sizes were also evaluated in alveolar lavages. The alveolar lavages recovered from aged rats contained a significantly higher proportion of LA than did those obtained from young or newborn rats. There was also an age-related decrease in the rate of conversion from LA to SA in vitro. The Sat-PC pool sizes in the alveolar lavages decreased with age, but the SP-A contents were similar between young and aged rats. These results suggested that decreased form conversion may contribute to maintaining functional surfactant pool sizes in the lungs of aged rats. PMID- 10678626 TI - Respiratory effects of lipopolysaccharide-induced inflammatory lung injury in mice. AB - The pathogenic mechanisms of lipopolysaccharide (LPS)-induced lung injury have not been classified. This study examined the physiological changes after endotoxin inhalation and related those to features of pulmonary inflammation in mice. Pulmonary mechanics, histopathology, and bronchoalveolar lavage fluid (BALF) from BALB/c mice were analysed at different occasions (3, 24, 48 and 72 h) after inhalation of saline or LPS from Escherichia coli (0.3 (L0.3) or 10 mg x mL(-1) (L10)). Mice were sedated, anaesthetized, and ventilated. After chest wall resection static (Est) and dynamic (Edyn) elastances, deltaE (Edyn-Est), resistive (deltaP1) and viscoelastic/inhomogeneous pressures (deltaP2), and deltaP1+deltaP2 (deltaPtot) were obtained by end-inflation occlusion method. Lungs were prepared for histopathology. In parallel groups, tumour necrosis factor (TNF)-alpha, neutrophils, and protein were evaluated in the BALF. L0.3 and L10 showed a time-dependent production of TNF-alpha preceding a massive neutrophil infiltration. In L10 BALF there was an increase in protein level at 24 and 48 h. Est and Edyn increased early in L0.3 (65%, 63%) and L10 (41%, 51%). In L10 deltaE, deltaP2, and deltaPtot showed a gradual rise. At 72 h all groups were similar. L0.3 showed an early increase in cellularity, which returned to normal at 72 h. L10 presented the same pattern with the cell count remaining elevated until 72 h. In conclusion, lipopolysaccharide inhalation led to elastic and viscoelastic pulmonary changes together with tumour necrosis factor-alpha production and neutrophil infiltration in mouse lung. PMID- 10678627 TI - Skeletal muscle strength and endurance in patients with mild COPD and the effects of weight training. AB - This study poses two questions: 1) is there an abnormality in isokinetic skeletal muscle strength and endurance in mild chronic obstructive pulmonary disease (COPD)? and 2) what is the effect of a randomized, controlled, 12 week hospital outpatient weight training programme in terms of skeletal muscle function and exercise tolerance? Upper and lower limb isokinetic maximum and sustained muscle function were compared in 43 COPD patients (age 49+/-11 yrs), mean forced expiratory volume in one second (FEV1) 77+/-23% pred and 52 healthy, sedentary subjects (age 51 (10) yrs), mean FEV1 109+/-16% pred. The 43 COPD patients were randomly allocated into training (n=26) and control (n=17) groups. Isokinetic and isotonic muscle function, whole body endurance, maximal exercise capacity and lung function were measured. The COPD patients had reduced isokinetic muscle function (with the exception of sustained upper limb strength) as compared with healthy sedentary subjects. Muscle function improved after weight training in the COPD patients. Whole body endurance during treadmill walking also improved with no change in maximal oxygen consumption. A deficit in skeletal muscle function can be identified in patients with mild chronic obstructive pulmonary disease which cannot be explained by factors such as hypoxaemia and malnutrition. Intervention with weight training is effective in countering this deficit which the authors conclude is probably due to muscle deconditioning. PMID- 10678628 TI - Nasal ventilation in COPD exacerbations: early and late results of a prospective, controlled study. AB - Noninvasive positive pressure mechanical ventilation (NIPPV) in exacerbated chronic obstructive pulmonary disease (COPD) has been investigated early and after 1 yr of follow-up. To this end, 30 patients were enrolled in a prospective, controlled trial: 15 had early administration of NIPPV (Group A), 15 had medical therapy only (Group B); assignment was made on the basis of equipment availability only. In-hospital mortality, need for endotracheal intubation and mean length of hospitalization were lower in Group A, though the difference was not statistically significant. Arterial oxygen tension in arterial blood (Pa,O2), carbon dioxide tension in arterial blood (Pa,CO2) and HCO3- improved significantly in both groups from admission to discharge: 45.8+/-8.6 versus 64.9+/-10.0; 59.4+/-11.8 versus 48.6+/-7.3; 34.3+/-4.3 versus 30.1+/-3.4 in group A; 49.2+/-11.4 versus 60.9+/-8.2; 52.6+/-15.9 versus 44.4+/-8.7; 31.7+/-5.9 versus 28.0+/-3.6 in group B, respectively, p<0.05 for all comparisons; pH, percentage forced expiratory volume in one second (FEV1) and tidal volume (VT) improved significantly in patients of group A only: 7.36+/-0.04 versus 7.41+/ 0.02; 39.8+/-13.6 versus 49.4+/-11.7; 0.71+/-0.3 versus 0.84+/-0.4, respectively, p<0.05. During follow-up, 3, 6, and 12 months survival rates were significantly higher in Group A than in Group B (p<0.02). Hospital new admissions over 1 yr were more frequent in Group B (n=6, incidence rate: 0.216%) than in Group A (n=4, incidence rate: 0.084%). Therefore, noninvasive positive pressure mechanical ventilation may be added to "conventional" medical therapy in exacerbated chronic obstructive pulmonary disease. PMID- 10678629 TI - Peripheral airway findings in chronic obstructive pulmonary disease using an ultrathin bronchoscope. AB - The authors performed bronchoscopic examination using an ultrathin bronchoscope to determine the characteristics of the peripheral airways in chronic obstructive pulmonary disease (COPD). The study population comprised 13 healthy control subjects, 10 patients with chronic bronchitis without airflow limitation, and 20 patients with COPD. The COPD patients were divided clinically into 10 with chronic bronchitis and 10 with pulmonary emphysema. The peripheral airways were examined using an ultrathin bronchoscope. In chronic bronchitis, peripheral airways of the 11th or 12th generation showed a high frequency of obstruction and mucosal changes such as granulation. In pulmonary emphysema, the peripheral airways frequently showed a net-like appearance of the bronchial epithelium and obstruction at the 11th or 12th generation. Morphological changes of the small airways in chronic obstructive pulmonary disease can be detected by an ultrathin bronchoscope, and this method is likely to be useful for investigating the small airways in vivo. PMID- 10678630 TI - Comparison of induced sputum with bronchial wash, bronchoalveolar lavage and bronchial biopsies in COPD. AB - It is unclear how cellular and soluble inflammatory markers in induced sputum relate to markers in lavage fluid and biopsies in chronic obstructive pulmonary disease (COPD). This was investigated and also the possible differences between subjects with COPD and healthy controls assessed. Eighteen nonatopic subjects with COPD and 11 healthy controls were studied. Sputum was induced by inhalation of hypertonic saline. The airways were lavaged, using the first 50 mL for bronchial wash (BW) and the subsequent 150 mL for bronchoalveolar lavage (BAL), and biopsies were taken from subsegmental carinae. Neutrophils were the predominant cell type in sputum in COPD (median 77.3%) but not in BW (5.5%) and BAL fluid (1.7%). Differential cell counts in sputum did not correlate with the counts in BW or BAL fluid or biopsies, whereas sputum eosinophil cationic protein (ECP) levels correlated with BW fluid ECP levels (p=0.66, p=0.007) and sputum interleukin-8 (IL-8) concentration with BAL fluid IL-8 concentration (p= 0.52, p=0.026). Subjects with COPD had a higher percentage of sputum neutrophils and eosinophils and higher concentrations of ECP and IL-8 than healthy controls. The higher percentages of eosinophils and concentrations of ECP were also seen in BW and BAL fluid. Finally, higher numbers of macrophages and eosinophils were found in biopsies. In conclusion, induced sputum is derived from a different compartment from BW and BAL fluid and biopsies. Induced sputum may be useful for studying the contribution of luminal neutrophils and eosinophils in chronic obstructive pulmonary disease. PMID- 10678631 TI - Venous and arterial changes in pulmonary veno-occlusive disease, mitral stenosis and fibrosing mediastinitis. AB - The pathogenesis of pulmonary veno-occlusive disease (PVOD) is not known. The diagnosis of PVOD frequently relies on its histological changes since it is often difficult to distinguish clinically from primary pulmonary hypertension. This study carried out a systematic analysis of the pulmonary venous and arterial remodelling that occurs in PVOD (n=5) and compared these changes to two other diseases affecting the pulmonary veins, mitral stenosis (MS; n=6) and fibrosing mediastinitis (FM; n=2), using established morphometric techniques. In PVOD, pronounced intimal and adventitial thickening were noted in veins of all sizes and arterialization of veins >50 microm external diameter was found. Similar changes were evident in the arterial wall, but intimal thickening was less severe than in the veins and medial thickening was more pronounced in arteries <300 microm external diameter. Eccentric intimal fibrosis of the veins was also noted for the first time in PVOD, although this feature occurred less frequently (approximately one third) than in MS. Less pronounced structural remodelling was also encountered in the veins in cases of MS and FM. The severity of the venous changes in PVOD may aid its diagnosis and lend insight into its pathogenesis. However, the similarity of the vascular changes in each form of venous hypertension also suggests that pathology alone may not always differentiate between these disease states. The similarity of the vascular changes in the three forms of venous hypertension suggests that, as in pulmonary artery hypertension, pressure, per se, is one of the triggers to vascular remodelling. PMID- 10678632 TI - Theophylline improves acute mountain sickness. AB - A randomized two-part study was conducted in order to determine the efficacy of theophylline in the treatment of acute mountain sickness during fast ascent to altitudes >2,500 m. Fourteen healthy male subjects participated in a randomized single-blind placebo-controlled crossover study carried out in a decompression chamber (simulated altitude 4,500 m). A second randomized single-blind, placebo controlled study was conducted at a high-altitude research laboratory (3,454 m) and included 21 healthy male subjects. The study medication was either 375 mg oral slow-release theophylline (250 mg if <70 kg) or a matched placebo tablet taken twice daily. The acute mountain sickness score (AMSS) was assessed three times a day, beginning 18 h prior to altitude exposure and continuing for 18 h after altitude exposure. In addition, measurements of respiratory frequency, pulse rate, oxygen saturation and arterial blood gas levels were performed. Acute mountain sickness was significantly reduced by theophylline during the decompression chamber study (mean+/-SD 1.2+/-0.9) with placebo versus 3.6+/-0.8 with theophylline; p=0.03). During the high-altitude study, subjects with theophylline showed a significantly lower AMSS on arrival and after 18 h at altitude (0.6 versus 2.3, p=0.03). Oxygenation was improved in both parts of the study. In conclusion, oral slow-release theophylline improves acute mountain sickness. PMID- 10678633 TI - Assessing the risk of hypoxia in flight: the need for more rational guidelines. AB - This study aimed to test the hypothesis that advice currently given by respiratory physicians to potentially hypoxic patients planning air travel varies and is not evidence-based. A prospective observational study was performed, surveying respiratory physicians in England and Wales. Sixty-two per cent responded. Nearly two-thirds worked in district general hospitals, a quarter in university hospitals, and the rest in tertiary referral (specialist) centres or a combination thereof. Most provide advice routinely; most of the remainder do on request or if concerned. Assessments comprise spirometry, blood gas level measurement, oximetry, predictive equations and hypoxic challenge tests. Twenty five per cent of physicians measuring blood gas levels recommend in-flight oxygen when arterial oxygen tension (Pa,O2) <7.3 kPa, 50% when Pa,O2 is 7.3-8.0 kPa. Over two-thirds using spirometry recommend oxygen when the forced expiratory volume in one second <40% of the predicted value. Half recommend oxygen when arterial oxygen saturation (Sa,O2) <90%, 33% when Sa,O2 is 90-94%. Fewer than 10% of district hospital physicians (and none in other hospitals) use predictive equations. More than half of specialists but fewer than 10% of district hospital physicians perform hypoxic challenge tests. The risk of hypoxia at altitude is recognized by most respiratory physicians in England and Wales, but assessment methods and criteria for recommending oxygen vary widely. This suggests that most current advice is not evidence-based. Evidence-based guidelines are required. PMID- 10678634 TI - Clinically "small" effects of air pollution on FVC have a large public health impact. Swiss Study on Air Pollution and Lung Disease in Adults (SAPALDIA) - team. AB - Epidemiological studies have repeatedly established adverse health effects due to long-term exposure to ambient air pollution. The Swiss Study on Air Pollution and Lung Disease in Adults (SAPALDIA) published a -3.14% decrease in forced vital capacity (FVC) per 10 microg x m(-3) increment in particulate matter (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM(10)). Compared to the within-subject variability of FVC, the effect may be considered small. This individual (or clinical) perspective is, however, misleading. The purpose of this study was to demonstrate the public health relevance of apparently "small" effects, using the impact of PM10 on FVC as an example. The scenario compares a population A, exposed to an annual mean PM10 of 20 microg x m(-3), with a population B exposed to 30 microg x m(-3) mean PM10. A shift of -3.14% in the population distribution of FVC increases the number of subjects in the lower tail of the distribution. In population B a relative increase was expected of 47% (16 91%) in the prevalence of "FVC <80% predicted", (i.e., from 5.17 to 7.59% and 5.88 to 8.65% among males and females, respectively). The relative increase in the prevalence of "FVC <70% predicted" (approximately 1% of population) was 63% (30-98%, males) and 57% (21-86%, females). An epidemiological estimate of a change in the mean value of the population distribution should not be misinterpreted as an effect on the individual level. However, the impact of a 10 microg x m(-3) increase in particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10) on the number of subjects with a clinically relevant reduction in lung function is quantitatively important. PMID- 10678635 TI - Impact of correcting peak flow for nonlinear errors on air pollutant effect estimates from a panel study. AB - Air pollutant effects are commonly investigated using panel studies employing daily measurement of changes in peak expiratory flow (PEF). Variable orifice PEF meters are inaccurate with a nonlinear relationship to actual PEF. The impact on a panel study of correcting these errors was examined. Twice-daily PEF readings were taken by 147 9-yr old children for 8 weeks and corrected using an equation derived from the response of 32 Vitalograph meters to a servomechanism-controlled pump. Pollutant effect estimates for corrected and uncorrected readings were derived using a regression approach incorporating appropriate confounders. Correction impacted little on mean PEF values (333.1-334.2 L x min(-1)), but did alter effect sizes. Nonsignificant nitrogen dioxide estimates for the entire panel decreased by up to 73%, but, for symptomatic/atopic children, a significant 5-day mean result was lost (decrease in effect size from -2.53 to -0.90% per 10 parts per billion (ppb)) and lag 0 became significant (decrease from -0.51 to 1.22% per 10 ppb). Mass concentration estimates of particles with a 50% cut-off aerodynamic diameter of 2.5 microm moved in both directions (-0.22 changed to 0.11% per 10 microg x m(-1) lag 3 and -0.29 to -0.73% per 10 microg x m(-3) for the 5-day mean). Correction of nonlinearity of peak expiratory flow meters influenced the overall outcome of this panel study, and the changes in effect estimates would be sufficient to alter the interpretation of some studies. For adults, a greater change in effect estimates may follow the larger correction required for their usual peak expiratory flow range. PMID- 10678636 TI - Fractional analysis of bronchoalveolar lavage fluid cytology in cystic fibrosis patients with normal lung function. Bronchoalveolar lavage for the evaluation of anti-inflammatory treatment (BEAT) study group. AB - Cystic fibrosis (CF) is associated with a neutrophil dominated airway inflammation. So far bronchoalveolar lavage (BAL) studies in CF have used pooled BAL samples which may be more representative of the alveolar compartment rather than the airways. To assess whether the first sample of a BAL is more sensitive in the evaluation of airway inflammation, the authors have studied 105 stable CF patients aged 5-37 yrs with a mean forced expiratory volume in one second (FEV1) of 96+/-15% (mean+/-SD). BAL cytology of the first and pooled samples were compared to reference values obtained in children without respiratory disease. Absolute cell counts and the percentage of neutrophils were significantly increased in CF patients. If the 95% confidence interval was used as a cut-off point, 17/105 CF patients had a normal percentage of neutrophils in pooled BAL samples, but only three also had a normal percentage of neutrophils in the first BAL aliquot. Therefore, neutrophil dominated airway inflammation is more pronounced in the first, mainly bronchial, bronchoalveolar lavage sample suggesting that sequential analysis of bronchoalveolar lavage fluid may have a higher sensitivity to detect early inflammatory changes in CF patients. PMID- 10678637 TI - Sodium channel blockers and uridine triphosphate: effects on nasal potential difference in cystic fibrosis mice. AB - Sodium channel inhibitors block the enhanced Na+ reabsorption in cystic fibrosis (CF). Extracellular nucleotides facilitate Cl- secretion via Ca2+ gated Cl- channels. A combination of these effects may produce less viscid secretions in CF which are easier to expectorate. This study examined the effects of combining sodium channel blockers with uridine triphosphate (UTP) on nasal membrane potential difference (PD) in CF insertional null mutant mice (cftr(tm1HGU)), deltaF508 homozygous mice (cftr(tm1Cam)) and matched control animals. Median basal PD in the insertional CF mice and deltaF508 CF mice were -28 and -34 mV respectively. These values were significantly different to the control animals ( 20 mV). Amiloride and loperamide reduced the PD in cftr(tm1HGU) CF mice (deltaPD 13 mV & 15 mV respectively) suggesting Na+ blockade. The subsequent addition of UTP in a chloride-free vehicle increased the PD (deltaPD -8- -12.5 mV). DeltaF508 mice showed significantly greater responses compared with CF insertional null mutant mice (p<0.05). The action of UTP was brief and not prolonged by the addition alpha-beta-methylene-adenosine 5' diphosphate. Suramin, a competitive antagonist of P2 purinoceptors blocked the action of UTP. In conclusion, this study demonstrated dose dependant nasal membrane potential changes in differences mice with uridine triphosphate in the presence of sodium channel blockers suggestive of chloride secretion. More stable analogues of uridine triphosphate in combination with long acting sodium channel blockers such as loperamide may have therapeutic potential in cystic fibrosis. PMID- 10678638 TI - The association between early life lung function and wheezing during the first 2 yrs of life. AB - Reports have suggested that certain infants are predisposed to wheezing in the first 2 yrs of life due to abnormal lung function, prior to the first wheezing illness. The authors investigated the association between infant lung function and wheeze during the first 2 yrs of life. A cohort of 253 infants was evaluated. Respiratory function assessment was performed at 1, 6, and 12 months of age. Parental history of asthma, atopy, and maternal antenatal smoking habits were recorded. An infant was identified as having wheezed on the basis of parental report and, where possible, physician diagnosis. One hundred and sixty infants (63%) had complete diary and questionnaire information on wheeze available for analysis. Of these: 79 infants (50%) had never wheezed (NW) during the first 2 yrs of life and 81 had reported wheeze (W) (50%). Of those with a report of wheeze, the distribution through the first 2 yrs of life was; 28 during the first year of life only (Y1), 21 in the second year of life only (Y2), and 32 wheezed in both the first and second years of life (Y1&2). At the age of 1 month, prior to any lower respiratory illness, the W group had impaired lung function in comparison to the NW group. In Y1 infants, the neonatal lung function differences resolved by 12 months of age. In Y2 and Y1&2 infants lung function differences persisted throughout the first year of life. Prevalence of parental asthma and maternal antenatal smoking was increased in the W group p=0.001, p=0.008, respectively), in comparison to the NW infants. Maternal antenatal smoking prevalence was increased in the Y2 and Y1&2 infants in comparison to the NW group (p=0.04), (p=0.01), respectively. Wheezing during the first year of life is often a transient condition which improves with time. It appears to be related to early life reduced small airway calibre. Wheezing that begins or persists into the second year of life is usually associated with a different abnormality of the airways. Commencement or persistence of wheeze into the second year of life may be part of the clinical entity recognized as asthma. PMID- 10678639 TI - Maturational changes of endothelial vasoactive factors and pulmonary vascular tone at birth. AB - The aim of this study was to determine which endothelial factors were involved in the decrease of pulmonary vascular resistance at birth, and how they changed with maturation. Response of intrapulmonary artery rings precontracted with prostaglandin F2alpha were studied from piglets aged <2 h, 2-3 day, 10 day and adult pigs for pharmacological responses to acetylcholine (ACh) and cromakalim (CMK) in the presence and the absence of the nitric oxide synthase (NOS) inhibitor, N(omega)-nitro-L-arginine (L-NA), the adenosine triphosphate sensitive potassium (K(ATP)) channel blocker, glibenclamide and the endothelin (ET)-A receptor antagonist, BQ123. In situ hybridization and immunochemistry studies were performed in lung tissues of the same animals in order to determine the expression of NOS and ET. There was a small contractile effect of ACh in the newborn. Relaxation to ACh, which was blocked by L-NA and reduced by glibenclamide, only appeared from the age of 3 days. The significantly greater relaxation to CMK in rings without endothelium (p<0.05) was abolished by BQ123 in the newborn, and then disappeared by 2 days of age. Glibenclamide had a greater inhibitory effect on relaxation induced by CMK at 10 days than in the newborn and 2 days old piglets. NOS expression was low in pulmonary arteries of the newborn and increased by 2 days of age whereas the converse was seen with ET expression. It is concluded that: 1) relaxant response to acetylcholine was absent at birth and appeared at 2 days; 2) the reduced relaxant response to cromakalin in rings with endothelium at birth could be blocked by BQ123; and 3) the expression of endothelin decreased whereas the expression of nitric oxide synthase increased from birth to 2 days of age. PMID- 10678640 TI - Diagnostic utility of eosinophils in the pleural fluid. AB - This study was conducted to assess the prevalence of eosinophilia in 358 consecutive samples of pleural fluid (all cases corresponded to first thoracentesis), to review the cause of eosinophilic pleural effusions, and to determine whether the presence of eosinophils increases the likelihood of nonmalignant underlying disorders. Eosinophilic pleural effusions were identified in 45 patients (12.6%): malignant underlying conditions were diagnosed in 11 patients (24.4% with eosinophilic effusions) and benign aetiologies were found in 27 patients. Benign aetiologies included uncomplicated paraneumonic effusion in 10 patients, tuberculosis in seven, complicated paraneumonic in five, liver cirrhosis in three, hydronephrosis in one and pulmonary thromboembolism in one. Seven pleural effusions were idiopathic. There was no difference in the prevalence between eosinophilic and noneosinophilic effusions according to the different diagnoses. With parameters of sensitivity, specificity, pretest and post-test probability and positive and negative predictive values for any prevalence figure using the Bayes' theorem and for any value of eosinophils (both in percentage or absolute numbers) in the pleural fluid (receiver operating characteristic curve) an adequate predictor of benign disease was not found. It is concluded that pleural eosinophilia at the initial thoracentesis cannot be considered as a predictor of an underlying benign disorder. PMID- 10678641 TI - Release of endothelins and platelet-activating factor by a rat pleural mesothelial cell line. AB - Thrombin is a multifunctional serine protease. It is generated in inflammatory processes and induces the proliferation and chemotaxis of a variety of cells including mesothelial cells (MTCs). MTCs are epithelial cells derived from the mesoderm, as are the vascular endothelial cells. Since thrombin acts on endothelial cells to produce platelet-activating factor (PAF) and endothelin (ET) 1, it was hypothesized that MTCs also produce PAF and ET via the action of thrombin. Rat pleural MTC (RMTC, 4/4 R.M.-4) monolayers were cultural in tissue culture dishes for various periods. The supernatants were fractionated by means of high-performance liquid chromatography to determine the ET isoforms and PAF species present. Immunoreactive ET was measured using an enzyme-linked immunosorbent assay, and PAF was measured by means of a bioassay using a platelet aggregometer. ET-1, ET-2 and ET-3 were detected in RMTC-conditioned medium, and the predominant isoforms were ET-1 and ET-2. RMTCs mainly released C16:0 PAF into the supernatant. Immunoreactive ET and PAF were released via the action of thrombin. Synthetic PAF significantly induced secretion of ET, but the PAF receptor antagonists, WEB2086 and E6123, failed to modulate thrombin-induced ET release. These results indicate that thrombin acts on pleural rat mesothelial cells to release ET and PAF, which may play a role in the development of pleurisy. PMID- 10678642 TI - The measurement of exhaled carbon monoxide is influenced by airflow obstruction. AB - The concentration of carboxyhaemoglobin (COHb) is often estimated from measurements of carbon monoxide in the exhaled air (COexh). This study investigates whether the presence of airflow obstruction significantly alters the relationship between COexh and COHb. Eighty-one regular smokers were prospectively studied and divided in four groups according to the presence and severity of airflow obstruction (none, mild, moderate, severe). In each subject, the authors measured in this order: 1) arterial blood gases; 2) haemoglobin concentration and COHb (by co-oxymetry); 3) COexh; 4) lung volumes; and 5) forced spirometry. The size of the measurement error (deltaCO) was calculated from the difference between COHb and COexh. Neither the smoking history nor COexh were different in the four groups of subjects studied. In contrast, deltaCO increased in parallel to the degree of airflow obstruction. DeltaCO was >2% (a threshold value normally used in the clinic to separate smokers from nonsmokers) only in patients with severe airflow obstruction. A stepwise multivariate analysis showed that both forced expiratory volume in one second (FEV1) (percentage reference) and COHb contributed significantly (p<0.0001) to predict deltaCO. This study shows that the estimation of carboxyhaemoglobin from exhaled carbon monoxide measurements can be inaccurate in patients with severe airflow obstruction. In these patients, the direct measurement of carboxyhaemoglobin seems advisable in clinical practice. PMID- 10678643 TI - Comparison of two standardized methods of methacholine inhalation challenge in young adults. AB - In the European Community Respiratory Health Study (ECRHS), airway responsiveness to methacholine was determined using the Mefar dosimeter protocol. Elsewhere, the 2-min tidal breathing method has become the preferred standardized method. The relationship between measurements of responsiveness by these two methods is not well established. This study measured airway responsiveness to methacholine by dosimeter and tidal breathing methods in 47 healthy asthmatic subjects aged 20-44 yrs. Tests were performed within 1 week and in random order. Baseline forced expiratory volume in one second (FEV1) varied by <10% between tests in 42/47 subjects. There was a close association between responsiveness determined by the two methods. A provocative concentration of methacholine causing a 20% fall in FEV1 (PC20) value of < or =8.0 mg x mL(-1) (tidal method) used to categorize airway hyperresponsiveness agreed most closely with a provocative dose of methacholine causing a 20% fall in FEV1 (PD20) value of < or =0.5 mg (dosimeter method) (kappa statistic 0.78). Each doubling or halving of PC20 to define a level of hyperresponsiveness agreed closely with a doubling or halving of PD20. Assessment of airway responsiveness as provocative dose of methacholine causing a 20% fall in forced expiratory volume in one second by the Mefar dosimeter protocol gave a close and predictable relationship with provocative concentration of methacholine causing a 20% fall in expiratory volume in one second assessed using the tidal breathing method. Airway hyperresponsiveness as determined by the accepted criterion of provocative concentration of methacholine causing a 20% fall in expiratory volume in one second < or =8 mg x mL(-1) was best correlated with provocative dose of methacholine causing a 20% fall in forced expiratory volume in one second <0.5 mg by Mefar dosimeter. PMID- 10678644 TI - A simplified method for monitoring respiratory impedance during continuous positive airway pressure. AB - The forced oscillation technique is useful in detecting changes in upper airway obstruction in patients with sleep apnoea undergoing continuous positive airway pressure (CPAP) ventilation. The aim of this study was to implement and evaluate a method for estimating respiratory impedance (Zrs) from the pressure and flow recorded at the inlet of the CPAP tubing. The method is based on correcting impedance measured at the inlet of the CPAP tubing (Zi) for the effect of the tubing and the exhalation port. The method was evaluated in mechanical analogues and in a healthy subject. Sinusoidal oscillation of 5, 10 and 20 Hz were superimposed on CPAP (5-15 cmH2O). At 5 Hz, the changes in airflow obstruction were substantially underestimated by Zi. Furthermore, Zi exhibited a negative dependence on Zrs at 20 Hz. The assessment of Zrs was greatly improved after correcting Zi for the effects of the CPAP tubing and the exhalation port. Zrs was well estimated at low frequencies, reaching very high values during total occlusion (>60 cmH2O x s x L(-1) at 5-10 Hz). These results indicate that changes in airflow obstruction can be detected using the forced oscillation technique from pressure and flow recorded on the continuous positive airway pressure device. This facilitates the clinical application of the forced oscillation technique for monitoring upper airway patency during sleep. PMID- 10678645 TI - Shortening the duration of treatment for cervical tuberculous lymphadenitis. AB - The aim of the study was to determine the optimal duration of treatment for patients with tuberculous lymphadenitis. The Medline database was searched for relevant articles published between 1978-1997. Inclusion criteria were study populations of patients with predominantly cervical tuberculous lymphadenitis in whom the diagnosis had been confirmed bacteriologically and/or histologically, or was made probable by using clinical and laboratory markers. Treatment management had to include at least isoniazid, rifampicin and pyrazinamide and a follow-up of at least 12 months after the end of treatment. Patients with resistance to rifampicin and pyrazinamide and previous treatment for tuberculosis were excluded. The number of patients who relapsed after treatment was calculated. The study population in eight out of the 35 articles retrieved were suitable for analysis. Some concerned comparative studies. There were eight treatment schedules of 6 months' duration and three schedules of 9 months' duration. Treatment for 6 months resulted in a tuberculous lymphadenitis relapse rate of 13/422=3.3% (95% confidence interval: 1.7-5.5), with a mean follow-up of 31 months after completion of treatment. Treatment for 9 months resulted in a relapse rate of 3/112=2.7% (95% confidence interval: 0.6-7.8), with a mean follow up of 20 months. Despite the limitations of the literature available, 6 months of therapy is probably sufficient for patients with tuberculous lymphadenitis. PMID- 10678646 TI - Physiotherapy for airway clearance in paediatrics. AB - The basic therapeutic principles in paediatric chest physiotherapy (CPT) are identical to those applied in adults. However, the child's growth and development results in continuing changes in respiratory structure and function, and the requirement for different applications of CPT in each age group. Forced expiratory manoeuvres and coughing serve as basic mechanisms for mobilization and transport of secretions, but the reduced bronchial stability after birth requires special techniques in very young patients. High externally applied transthoracic pressures have to be avoided in order to prevent interruption of airflow. In addition, airway patency is maintained by the application of back pressure and by liberal use of continuous positive airway pressure. Since sympathomimetic bronchodilators might further decrease bronchial stability, their use must be individualized in newborns and young infants. Inspiration is a basic mechanism for inflating alveolar space behind obstructing mucus plugs. Due to a highly unstable chest, the premature baby, newborn and infant cannot distend their lung parenchyma to the same extent as can older patients. Consequently all chest physiotherapy strategies applied in this age group have to incorporate appropriate techniques for raising lung volume. Positioning serves to redistribute ventilation, but the young infant's response to gravitational forces differs substantially from that of the adult, and consequently strategies used in older patients have to be modified. In addition, the therapist has to consider pathology such as bronchial instability lesions and airway hyperresponsiveness and has to adjust the therapeutic response accordingly. It is particularly important to consider the special vulnerability of newborns and young infants and to modify therapeutic interventions to avoid the harm that could otherwise be inflicted. Consideration of these differences between infant, child and adult and careful analysis of the available mucus clearance techniques allows tailoring of an individualized therapeutic approach to the paediatric patient. PMID- 10678647 TI - Eosinophilic pneumonia and respiratory failure associated with venlafaxine treatment. AB - Drugs are well known causes of eosinophilic lung disease. In many patients, symptoms increase slowly, pulmonary infiltrates and eosinophilia progress over weeks, and resolve upon withdrawal of the offending agent. Rarely, the disease presents like acute eosinophilic pneumonia with acute onset of symptoms and rapidly progressing infiltrates which may be associated with respiratory failure. This report describe a case of venlafaxine-induced acute eosinophilic pneumonia causing respiratory insufficiency that rapidly resolved upon institution of corticosteroid treatment. This 5-hydroxytryptamine and noradrenaline reuptake inhibitor was previously not known to cause lung or peripheral blood eosinophilia. Considering the increasing use of this class of medication physicians have to be aware of this life-threatening and fully reversible complication. PMID- 10678648 TI - Severe tracheobronchial stenosis in a patient with Crohn's disease. AB - Tracheobronchial involvement in Crohn's disease is rare, usually associated with symptoms of tracheobronchitis, and typically responds well to steroids. The authors report a case of a 29-yr old patient with Crohn's disease, who presented with dyspnoea, fever, and a productive cough. Computed tomography of the chest revealed extensive nodular tracheobronchial stenosis, that was accompanied by severe mucosal inflammation at bronchoscopy. High-dose oral steroids diminished the mucosal inflammation, but had limited efficacy on the underlying tracheobronchial stenosis. It is speculated that this relative ineffectiveness of steroids may be due to the persistence of the untreated inflammatory process. PMID- 10678649 TI - Granulomatous Pneumocystis carinii pneumonia in Wegener's granulomatosis. AB - This study reports on a first case of granulomatous Pneumocystis carinii pneumonia (PCP) in a human immunodeficiency virus-negative patient with antineutrophil cytoplasmic antibody-positive Wegener's granulomatosis whilst receiving immunosuppressive treatment. The patient presented with diffuse alveolar haemorrhage, pauci-immune rapid progressive glomerulonephritis and leukocytoclastic vasculitis of the skin. Granulomatous Pneumocystis carinii pneumonia developed under immunosuppressive treatment with cyclophosphamide and prednisone. At the time Pneumocystis carinii pneumonia developed, there was a marked lymphopenia with a very low CD8+ cell count in the blood. Grocott staining in bronchoalveolar lavage fluid revealed no Pneumocystis carinii. The diagnosis was made via a video-assisted thoracoscopic lung biopsy which showed granulomas containing high numbers of Pneumocystis carinii cysts. PMID- 10678650 TI - Bronchoalveolar lavage in children. ERS Task Force on bronchoalveolar lavage in children. European Respiratory Society. PMID- 10678651 TI - Alveolar ejection volume: a misnomer? PMID- 10678652 TI - Difficult asthma. PMID- 10678653 TI - New syndrome?: Three sibs diagnosed prenatally with situs inversus totalis, renal and pancreatic dysplasia, and cysts. AB - Recently we described a previously apparently undescribed autosomal recessive syndrome in two sib fetuses with situs inversus totalis, cystic dysplastic kidneys and pancreas, bowing of the lower limbs and clavicles, severe intrauterine growth retardation, and oligohydramnios. This syndrome differs from that of Ivemark and related syndromes due to lack of liver involvement. After these two sibs, this consanguineous family had a third child and an early prenatal diagnosis of pancreatic and dysplastic renal cysts was made in the 19.5 week-old fetus. The last case supports the genetic hypothesis. PMID- 10678654 TI - Short stature in carriers of recessive mutation causing familial isolated growth hormone deficiency. AB - Isolated growth hormone deficiency (IGHD) IB is an autosomal recessive disorder characterized by a good response to exogenous growth hormone (GH) treatment without development of anti-GH antibodies. Patients with IGHD IB were found to be compound heterozygotes for deletion and frameshift mutations as well as homozygotes for splicing mutations in the GH-1 gene. Recently, a novel splicing mutation in the GH-1 gene was identified in an extended, consanguineous Arab Bedouin family from Israel with IGHD IB. Prior to the identification of this mutation, a considerable number of children with short stature in this family were found normal on pharmacological stimulation for GH release. This observation prompted a genotype/phenotype correlation of potential heterozygotes in the family. Carriers of the mutant GH-1 allele were found as a group to have a significantly shorter stature than normal homozygote (mean standard deviation scores, 1.67 and -0.40, respectively, P<0.05). Moreover, 11 of 33 (33%) heterozygotes, but only 1 of 17 (5.9%) normal homozygotes, had their height at 2 or more SD below the mean. Overall, 48.5% of studied heterozygotes were found to be of appreciably short stature with height at or lower than the 5th centile (> or = -1.7 SD), whereas only 5.9% of the normal homozygotes did (P<0.004). This phenomenon of heterozygotes for a recessive mutation in the GH-1 gene manifesting short stature, might imply that some such mutations may account for non-GH deficiency reduced height in the general population. PMID- 10678655 TI - Congenital absence of permanent teeth in a six-generation Chinese kindred. AB - We report on rare, heritable, permanent tooth agenesis in a large Chinese kindred. The congenital absence of permanent teeth except the first and second accessory teeth was observed in 52 individuals through six successive generations in the kindred comprising 328 members. Clinical assessments were carried out, and inheritance mode and spousal influence of the anomaly on their offspring were analyzed. Consequently, the anomaly was transmitted in an autosomal dominant fashion with incomplete penetrance (P = 0.88), and no significant clinical manifestations other than the oligodontia were found. A geographical or environmental effect on the affected individuals was obviously eliminated, because any who are related to the kindred but live under the same conditions are fully healthy. The disorder we describe, therefore, differs from any previously reported oligodontia/anodontia syndromes. The oligodontia ranged from a few teeth to the whole set of teeth, and usually occurred at a period from age 7 or 8 years, the time when primary teeth are normally replaced by permanent teeth, to the forties. Roentgenography of the affected persons indicated that only the first and/or second accessory teeth with tooth buds developed as permanent teeth. In fact, the diphyodontic germination sometimes occurred in the oral cavity of the affected individuals. PMID- 10678656 TI - Spondylometaphyseal dysplasia: Sedaghatian type. AB - We report a case of spondylometaphyseal dysplasia in an infant who was born to nonconsanguineous Yemeni parents. Radiological findings were consistent with lethal metaphyseal chondrodysplasia (Sedaghatian type). Although all previously reported cases died within 4 days of life, our patient survived 161 days. This reported case was thoroughly investigated for serum calcium, magnesium, zinc, ammonia, phosphate level, alkaline phosphatase, parathormone level, liver and renal function test, TORCH, metabolic screening, skeletal survey, chromosomal studies, muscle enzymes, EEG, computed tomography scan, and magnetic resonance imaging (brain). Genomic DNA analysis of patient and parents were sent to the Faculty de Medicine Xavier Bichat, France, but yet abnormal gene could not be detected. PMID- 10678657 TI - Cri du chat syndrome: changing phenotype in older patients. AB - The cri du chat syndrome or 5p deletion syndrome is a well-delineated clinical entity and has an incidence of 1/50,000 in newborn infants. A de novo deletion is present in 85% of the patients. Ten to 15% are familial cases with more than 90% due to a parental translocation and 5% due to an inversion of chromosome 5. Although the size of the deleted segment varies, the critical segment that is deleted in all patients appears to be 5p15.2. The clinical picture is well known in younger patients and includes the typical high-pitched cry, psychomotor retardation, microcephaly, growth rate failure, and craniofacial abnormalities including round face, hypertelorism, broad nasal bridge, downward slanting palpebral fissures, and micrognathia. With advancing age, the clinical picture becomes less striking. We present seven patients with 5p deletion syndrome, who were between age 16 and 47 years. Comparing their phenotype at several ages, a change of their phenotype was noted. Some of the clinical characteristics became more evident such as long face, macrostomia, and scoliosis. All patients were severely or profoundly mentally retarded except one patient who was mildly mentally retarded. The diagnosis was difficult to make in some of the patients who were first seen at an older age. In some of them, the craniofacial appearance resembled that seen in Angelman syndrome. Most patients had periods of destructive behavior, self mutilation, and aggression. The clinical diagnosis should be confirmed as soon as possible with cytogenetic investigation to provide specific support, prevention, and treatment of complications. Therefore, it is important to perform follow-up studies in young children to determine their outcome after infant-stimulation programs. PMID- 10678658 TI - Splicing mutations in the COL3 domain of collagen IX cause multiple epiphyseal dysplasia. AB - We report on a three-generation family with multiple epiphyseal dysplasia (MED). The propositus had typical MED findings of knees, ankles, elbows, and hands in childhood. The 2 other affected relatives were adults. The main clinical findings consisted of osteochondritis dissecans and osteoarthritis of the knees. DNA of the propositus was screened for mutations by conformation sensitive gel electrophoresis in all known candidate genes for MED, cartilage oligomeric matrix protein, and the COL9A1, COL9A2, and COL9A3 genes coding for the alpha1, alpha2, and alpha3 chains of collagen IX. The screening identified a unique change in PCR products of exon 3 of the COL9A3 gene. Sequencing indicated a G to A mutation in the acceptor splice site (G(-1)IVS2-->A) of intron 2 in all affected relatives, but not in unaffected relatives. Analysis of RNA from the propositus indicated a skipping of exon 3, and thus, a deletion of 12 amino acid residues as a consequence of the mutation. All four other collagen IX mutations previously described in MED have consequences identical to that characterized here, thus it seems likely that this type of mutation in collagen IX plays an important role in the pathogenesis of MED. PMID- 10678659 TI - Rothmund-Thomson syndrome due to RECQ4 helicase mutations: report and clinical and molecular comparisons with Bloom syndrome and Werner syndrome. AB - Rothmund-Thomson syndrome (RTS), an autosomal recessive disorder, comprises poikiloderma, growth deficiency, some aspects of premature aging, and a predisposition to malignancy, especially osteogenic sarcomas. Two kindreds with RTS were recently shown to segregate for mutations in the human RECQL4 helicase gene. We report identification of a new RTS kindred in which both brothers developed osteosarcomas. Mutation analysis of the RECQL4 gene was performed on both brothers and both parents. The brothers were shown to be compound heterozygotes for mutations in the RECQL4 gene, including a single basepair deletion in exon 9 resulting in a frameshift and early termination codon and a base substitution in the 3-prime splice site in the intron-exon boundary of exon 8, which would be predicted to cause a deletion of at least part of a consensus helicase domain. Each parent was shown to be a heterozygote carrier for one mutation. This report strengthens the association between mutations in RECQL4 helicase gene and RTS. Two other recessive disorders, Bloom syndrome and Werner syndrome, are known to be due to other human RECQ helicase gene mutations. These three disorders all manifest abnormal growth, premature aging, and predisposition to site-specific malignancies. The clinical and molecular aspects of RTS, Bloom syndrome, and Werner syndrome are compared and contrasted. PMID- 10678660 TI - Twin carriers of X-linked agammaglobulinemia (XLA) due to germline mutation in the Btk gene. AB - We report on an X-linked agammaglobulinemia (XLA) family in which mothers of two affected cousins were monozygotic twins. We analyzed the Btk gene of several members in three generations of the family by SSCP analysis, DNA sequencing, and RFLP analysis following polymerase chain reaction-amplification of the individual exons. We identified a missense point mutation, G1817C (R562P), in exon 17 of the Btk gene in the affected cousins. The same mutation was also present in both mothers (twin sisters) of the cousins identifying them as carriers. However, the mutation was absent in all other relatives including the grandmother of the cousins (mother of the twin sisters). This strongly suggests that the mutation in the Btk gene had originated in one of the germ lines or in the zygote. This may be the first demonstration of a germ line (or zygotic) mutation in XLA. PMID- 10678661 TI - Routine prenatal diagnosis of aneuploidy by FISH studies in high-risk pregnancies. AB - This study is a prospective clinical trial with fluorescent in situ hybridization (FISH) as a "routine" test for prenatal detection of the most common aneuploidies in high-risk pregnancies. Since April 1996, FISH studies with multicolor, commercially available, specific probes for chromosomes 13, 18, 21, X, and Y have been routinely performed in our cytogenetic laboratory on uncultured chorionic villous samplings (CVS), amniotic fluid samples, or fetal blood obtained by cordocentesis from patients with major or minor fetal anomalies detected by ultrasonography. Among the 4,193 prenatal samples analyzed between April 1996 and June 1998, routine FISH studies were ordered by the referring physicians on 301 (7.2%) cases. Aneuploidies were detected in 32 (10.6%) samples. Fourteen trisomy 21, 10 trisomy-18, 3 trisomy-13, 4 monosomies of X, and 1 case of triploidy were diagnosed by FISH. All 1,505 hybridizations were informative, and all 301 results were available and reported to the referring physicians in 24-48 hr. All relevant FISH results were confirmed by subsequent cytogenetic analysis. In 10 (3.8%) cases with normal FISH results, the final cytogenetic analysis revealed abnormal chromosomal rearrangements that could not be detected by the routine FISH studies. We conclude that rapid FISH analysis of interphase, uncultured fetal cells is an accurate and very sensitive method for routine prenatal diagnosis of the most common aneuploidies in high-risk pregnancies. PMID- 10678662 TI - Boy with syndactylies, macrocephaly, and severe skeletal dysplasia: not a new syndrome, but two dominant mutations (GLI3 E543X and COL2A1 G973R) in the same individual. AB - An unusual combination of syndactylies, macrocephaly, and severe skeletal dysplasia was observed in a newborn infant. A history of digital anomalies in the father and grandfather lead to the diagnosis of dominantly inherited Greig cephalopolysyndactyly syndrome (GCPS, MIM #175700). Having explained the digital findings and macrocephaly, the skeletal changes were thought to fit best congenital spondyloepiphyseal dysplasia (SEDC MIM #183900), a type II collagen disorder. Molecular analysis confirmed the presence of two dominant mutations in the propositus: a GLI3 mutation (E543X), which was present also in the father and grandfather, and a de novo COL2A1 mutation leading to a G973R substitution. Thus, this boy combined the syndactyly-macrocephaly phenotype of Greig cephalosyndactyly syndrome with a severe form of spondyloepiphyseal dysplasia caused by the structural defect in type II collagen. The diagnostic difficulties posed by the combination of two genetic disorders and the contribution of molecular diagnostics are well illustrated by this case. PMID- 10678663 TI - Giant congenital aortic aneurysm with cleft sternum, supraumbilical raphe, and hemangiomatosis: report and review. AB - We report on a child with giant congenital aortic aneurysm, sternal defect, hemangiomas of face, supraumbilical raphe, and review the only two other cases reported to date. Congenital aortic aneurysm is an ominous malformation that has to be systematically searched in children with the sternal malformation/vascular dysplasia complex. PMID- 10678664 TI - Pathogenetic classification of a series of 27,145 consecutive infants with congenital defects. AB - We studied a series of 27,145 consecutive infants with congenital defects and classified them into the currently recognized pathogenetic types of errors of morphogenesis, as defined by the International Working Group [Spranger et al., 1982: J Pediatrics 1:160-165]. Of all infants with congenital defects, 97.94% had malformations, 3.92% deformations, and 1.65% disruptions. Malformations associated with deformations were present in 3.12% of children with congenital anomalies, malformations with disruptions in 0.18%, deformations with disruptions in 0.07%, and malformations with deformations and disruptions in 0.14%. While deformations, including deformation sequences, were 2.38 times more common than disruptions and disruption sequences, isolated disruptions (1.27%) were more frequent than isolated deformations (0.59%). Knowledge of the frequencies of the different types of errors of morphogenesis (malformations, deformations, disruptions, developmental field defects, associations, complexes, unrecognized patterns of multiple congenital anomaly, and syndromes) may be of great value in the evaluation of patients with congenital anomalies. PMID- 10678665 TI - Recurrence risk for sibs of children with "sporadic" achondroplasia. AB - Because of gonadal mosaicism, the risk of recurrence of achondroplasia in the sibs of achondroplastic children with unaffected parents is presumably higher than twice the mutation rate, but it has not been measured. Data from 11 Canadian genetics centers provide an estimate of 1/443, or 0.02%. PMID- 10678666 TI - Ataxia-pancytopenia syndrome. AB - We report on a Mexican girl who developed cerebellar ataxia at age 3 years and pancytopenia at age 13 years. Cerebral computed tomography scan and magnetic resonance imaging showed evidence of severe cerebellar atrophy. Telangiectasias were not present; immunoglobulins and alpha-fetoprotein levels were normal. Cytogenetic studies showed no evidence of spontaneous chromosome aberrations, a normal rate of diepoxybutane (DEB) and mitomycin C (MMC)-induced chromosome aberrations, but an increased response to bleomycin. The phenotype support the diagnosis of ataxia-pancytopenia syndrome, although monosomy of chromosome 7 was not found in bone marrow. The cytogenetic studies suggest that this may be a chromosomal instability disorder. PMID- 10678667 TI - Whistling face syndrome with normal hands. PMID- 10678668 TI - Second case of bladder carcinoma in a patient with Costello syndrome. PMID- 10678669 TI - Incidence of Smith-Lemli-Opitz syndrome in Slovakia. PMID- 10678670 TI - Miscarriage and use of multi-vitamins or folic acid. PMID- 10678671 TI - The methods to measure stable isotope tracers and their applications to clinical nutrition problems keep getting better... PMID- 10678672 TI - Energy expenditure in wasting diseases: current concepts and measurement techniques. AB - Weight loss occurs secondary to many acute and chronic disease states. The measurement of energy balance is crucial to understanding disease-related changes in body composition. This review considers various techniques for measuring energy expenditure in wasting diseases and the current concepts regarding the effect of disease on energy expenditure. PMID- 10678673 TI - Neutron activation analysis determination of body composition. AB - The nutritional status of patients can be evaluated by monitoring changes in body composition, including the depletion of protein and muscle, adipose tissue distribution and changes in hydration status, bone or cell mass. Neutron activation analysis is a unique reference tool for the in-vivo determination of body composition. In this review we describe the recent changes in the field that followed the advent of new portable generators of fast neutrons, capable of performing elemental analysis in the clinical environment. New models were developed based on the partition of the measurable elements of the body. The recent developments help evaluate new treatments for wasting and obesity, in which change in body composition is the main outcome. PMID- 10678674 TI - Use of gas chromatography-combustion-isotope ratio mass spectrometry in nutrition and metabolic research. AB - Linking gas chromatography via an on-line combustion interface to isotope ratio mass spectrometry has opened the door to high-precision compound-specific isotope analysis. For this reason, gas chromatography-combustion-isotope ratio mass spectrometry is now increasingly employed in metabolic and nutritional research because it offers a reliable and risk-free alternative to the use of radioactive tracers. PMID- 10678675 TI - Inter-organ fluxes in a methodological perspective. AB - Inter-organ fluxes remain a cornerstone as an investigating tool in metabolic research. Combination with isotopic labelling and microdialysis have now opened new possibilities of addressing a number of questions not previously accessible. However, the technique requires a high level of knowledge among investigators as well as among readers. PMID- 10678676 TI - Plasma cytokines: what we are measuring. AB - Immunoassays, in particular enzyme-linked immunosorbent assays, have become increasingly important tools for the measurement of plasma cytokines. However, many technical factors contribute to the complexity of their quantitation. The study of plasma cytokine levels is also of limited value, and complementary methods are now available to investigate and understand more precisely the involvement of cytokines in pathological processes. PMID- 10678677 TI - Methods for assessing the potential of prebiotics and probiotics. AB - Prebiotics and probiotics are microflora management tools designed to improve human health. Both are dietary materials that fortify components of the gut flora seen as 'beneficial'. Gut flora modulation is an important area of the nutritional sciences, however, it is imperative that reliable methodologies be used to determine efficacy. This review will discuss the current techniques used in prebiotic and probiotic research. PMID- 10678678 TI - Modulation of endogenous glutathione availability. AB - Glutathione is quantitatively the most important antioxidant and scavenger. In addition it has a number of important functions in amino acid transport across membranes, in protein synthesis and degradation, in gene regulation and in cellular redox regulation. It becomes more and more evident that depletion of glutathione is associated with states of severe diseases. From this perspective, the possibility of manipulating the availability of glutathione becomes a very attractive form of treatment. In this review, new insights into pathophysiology and the regulation of glutathione metabolism, in addition to effects of precursors to and stimulants of glutathione synthesis, are covered. It is very likely that glutathione precursors will soon be an important pharmacological tool for treatment in a number of diseased states. PMID- 10678679 TI - N-acetyl-cysteine in the therapy of HIV-positive patients. AB - Randomly selected asymptomatic HIV-positive persons reveal, on average, a massive daily loss of sulphur, which appears to represent in first approximation the mean loss throughout the asymptomatic stage, and may explain the widely observed decrease in cyst(e)ine and glutathione levels. This sulphur loss is reasonably expected to lead, within a few years, to a life-threatening condition and may, therefore, contribute decisively to disease progression. Importantly, the rate of sulphur loss is not ameliorated by highly active antiretroviral therapy and may contribute to antiretroviral treatment failure. Several clinical trials on N acetyl-cysteine treatment of HIV-positive patients have revealed various therapeutic effects, but did not meet the rigorous standards for approval by the health authorities. PMID- 10678680 TI - Recent advances in conjugated linoleic acid research. AB - New results on the physiological properties of conjugated linoleic acid have been published by several working groups, especially showing the effects of single conjugated linoleic acid isomers on carcinogenesis and body composition. Recently, other studies have shown that conjugated linoleic acid has an influence on diabetes mellitus, platelet aggregation and the immune system. Conjugated linoleic acid was found to modify prostaglandin metabolism and delta9-desaturase activity and influence apoptosis. Furthermore, improved analytical methods using 13C nuclear magnetic resonance and silver ion high performance liquid chromatography are available to investigate the composition of conjugated linoleic acid mixtures and the exact structure of separated isomers. Also, the synthesis of isolated isomers is described, as published by different authors, in order to determine further the effects of each single conjugated linoleic acid isomer. In addition, new data on the contents of conjugated linoleic acid in foods, human adipose tissue and fluids are given in this review. More data need to be obtained using isolated isomers, with particular emphasis on studies in humans. PMID- 10678681 TI - Benefits and dangers of iron during infection. AB - Iron is essential for both human and microbial metabolism, and it is therefore important that iron status is maintained at a level that permits optimal immune responses by the host but does not facilitate availability of iron to microorganisms. This paper reviews the role of iron in resistance to infection, with particular reference to malaria and hepatitis C, and discusses the desirability of iron supplementation of populations at risk of infection. PMID- 10678682 TI - Carbohydrate and exercise. AB - Total body carbohydrate stores are limited, and are often less than the carbohydrate requirements of athletic training and competition. However, the availability of carbohydrate as a substrate for muscle metabolism is a critical factor in the performance of both high-intensity intermittent work and prolonged aerobic exercise. The rate of carbohydrate oxidation during exercise is tightly regulated, with glucose availability closely matching the needs of the working muscles. Both the absolute and relative work rate play important roles in the regulation of substrate metabolism: carbohydrate-based fuels predominate at moderate to high power outputs, with muscle glycogen and glucose utilization scaling exponentially to the relative work rate. As such, strategies to maintain or enhance carbohydrate availability, such as the ingestion of carbohydrate before, during and after exercise, are critical to the performance of a variety of sports events, and are a key recommendation in current sports nutrition guidelines. PMID- 10678683 TI - Dietary fat and physical performance. AB - Although the relationship between dietary carbohydrate and physical performance is well described, there is much controversy about the relationship between dietary fat and physical performance. Recently, several studies have tried to clarify this relationship. Here the effects of acute fat on metabolism and performance will be discussed, as well as the effects of short-term and long-term high-fat diets. PMID- 10678684 TI - The tricarboxylic acid cycle in human skeletal muscle: is there a role for nutritional intervention? AB - The tricarboxylic acid (TCA) cycle is essential for oxidative energy production. The expansion (anaplerosis) of the intermediates of the TCA cycle is achieved via a number of pathways, and is known to be influenced by metabolic status and nutritional and pharmacological interventions. Contraction is associated with anaplerosis in skeletal muscle, and some authors have suggested that the rate of anaplerosis can limit oxidative energy delivery. The results of more recent studies, however, are consistent with the idea that expansion of the muscle TCA intermediate pool is principally a reflection of muscle pyruvate availability, and is of little functional importance to TCA cycle flux, thereby indicating that any intervention aimed at increasing TCA intermediates expansion will be of little practical value. PMID- 10678685 TI - Protein and physical performance. AB - Important advances have been made in the understanding of the regulation of protein metabolism, which are of relevance to those interested in maximizing muscle performance in sport and aging. The use of 24 h amino acid infusion studies, the development of methods to measure skeletal muscle fractional breakdown rate and the fractional synthetic rate of specific proteins have and will continue to enhance our understanding of protein metabolism in exercise. Recent studies have demonstrated potentially beneficial effects upon protein metabolism by altering the composition and timing of nutrient delivery. Exercise and nutritional interventions may positively influence the negative effects of aging upon protein metabolism. PMID- 10678686 TI - Amino acid supplements to improve athletic performance. AB - This review provides a critical evaluation of the metabolic rationale for the use of individual amino acids as nutritional ergogenic (work-generating) aids in athletes. The conclusion is that in contrast to the claims made on sport nutrition products, branched-chain amino acids do not improve endurance performance, that the evidence that glutamine supplements may improve immune function is rather weak, and that the available commercial supplements contain too little arginine to increase growth hormone levels. No studies have been performed to investigate the claim that tyrosine supplements can improve explosive exercise. PMID- 10678687 TI - Bibliography. Current world literature. Assessment of nutritional status and analytical methods. PMID- 10678688 TI - Bibliography. Current world literature. Nutraceutics. PMID- 10678689 TI - Bibliography. Current world literature. Nutrition and physiological function. PMID- 10678690 TI - The effect of variability of unattended information on global and local processing: evidence for lateralization at early stages of processing. AB - Visual objects can often be analyzed as hierarchical in structure, composed of local elements that are spatially arranged to form a global shape. The brain mechanisms involved in the analysis of hierarchical figures have been under considerable scrutiny in recent years, and one of the many interesting features that have emerged is that there is an asymmetry across the two hemispheres for global (right hemisphere) vs local (left hemisphere) processing. Event-related potentials (ERP) were used to examine selective attention to global or local levels of hierarchical figures to determine the stage of processing at which the asymmetry first emerges. Two conditions were tested, one in which unattended information was variable from trial to trial, and one in which it was not. The variability of unattended information influenced the lateralization of processing. Presentation of invariable, neutral distractors resulted in global/local processing asymmetries at early stages (P1). In contrast, presentation of variable, task-relevant distractors resulted in processing asymmetries that occurred at much later stages (N2). Our hypothesis is that lateralized enhancement of neural populations in extrastriate cortex results from both selective attention to locations in the visual field, as well as selective attention to global or local information. We suggest that unattended information that varies from trial to trial is processed in parallel with attended information, masking hemisphere biases for local vs global information at early stages of processing. PMID- 10678691 TI - Implicit and explicit memory: a functional dissociation in persons with Down syndrome. AB - This study aimed at investigating implicit and explicit long-term memory functioning in subjects with Down syndrome (DS) compared to Mental-Age (MA) matched normal children. For this purpose, tests of verbal and visuo-perceptual explicit memory, verbal and visual repetition priming and procedural learning tasks were administered to 14 DS and 20 MA subjects. Our results document comparable implicit memory abilities in the two groups. In contrast, regarding explicit memory, normal children performed better than DS individuals. These results reveal a functional dissociation between implicit and explicit memory in subjects with DS. Theoretical and rehabilitative implications are discussed. PMID- 10678692 TI - The nature and staging of attention dysfunction in early (minimal and mild) Alzheimer's disease: relationship to episodic and semantic memory impairment. AB - The development of cholinergic therapies for Alzheimer's disease (AD) has highlighted the importance of understanding the role of attentional deficits and the relationship between attention and memory in the earliest stages of the disease. Variability in the tasks used to examine aspects of attention, and in the disease severity, between studies makes it difficult to determine which aspects of attention are affected earliest in AD, and how attentional impairment is related to other cognitive modules. We tested 27 patients in the early stages of the disease on the basis of the MMSE (minimal 24-30 corresponding to minimal cognitive impairment, very mild or possible AD in other classifications; and mild 18-23) on a battery of attentional tests aimed to assess sustained, divided, and selective attention, plus tests of episodic memory, semantic memory, visuoperceptual and visuospatial function, and verbal short-term memory. Although the mildly demented group were impaired on all attentional tests, the minimally impaired group showed a preserved ability to sustain attention, and to divide attention based on a dual-task paradigm. The minimally demented group had particular problems with response inhibition and speed of attentional switching. Examination of the relationship between attention and other cognitive domains showed impaired episodic memory in all patients. Deficits in attention were more prevalent than deficits in semantic memory suggesting that they occur at an earlier stage and the two were partially independent. Impairment in visuoperceptual and visuospatial functions and verbal short-term memory were the least common. Although attention is impaired early in AD, 40% of our patients showed deficits in episodic memory alone, confirming that amnesia may be the only cognitive deficit in the earliest stages of sporadic AD. PMID- 10678693 TI - Hemispheric asymmetries in the resolution of lexical ambiguity. AB - The linguistic phenomenon of lexical ambiguity has been intensively investigated as a means of gaining insight into general mechanisms of lexical access. It is now evident that both context and meaning frequency are significant factors in the determination of lexical outcomes. This suggests that hemispheric processes may be relevant to the resolution of lexical ambiguity, because both factors have been shown to have differential implications for the processing of language in the hemispheres. This study set out to examine the effects of context and meaning frequency on the resolution of ambiguous word meanings within the hemispheres. Sentences presented at the beginning of each trial embodied contexts which expressed either the dominant or subordinate meaning of a terminating homographic prime. Laterally presented target words reflected senses of the prime which were either consistent with, or inconsistent with, the context created by the preceding sentence. The most interesting results were observed at short prime target intervals where it was found that although dominant meanings of the target did not give rise to visual field differences, subordinate meanings evoked facilitated responses only in the left visual field. This result suggests that the right hemisphere immediately and exhaustively activates the various meanings associated with a word, while in the left hemisphere initial access is selectively restricted to the dominant meaning. It is proposed that this reflects a model of language comprehension in which the right hemisphere plays a supportive role by making available a set of alternative and less probable word meanings, thus freeing the left hemisphere to focus cognitive resources upon the most probable meaning of a word in a given context. PMID- 10678694 TI - Divided visuo-spatial attention systems with total and anterior callosotomy. AB - The role of the corpus callosum in the inter-hemispheric integration of the visuo spatial attention system, was investigated in patients with a total callosotomy or with an anterior callosal section. Subjects produced simple reaction times (RTs) to visual targets shown to the left or right visual hemifield. Preceding the target by an interval of 500 ms, arrow cues predicting the target location were shown left and right of the point of ocular fixation. For a majority of total and anterior callosotomy patients, results with valid focused cues (both arrows pointing to the target location) and with divided-attention cues (arrows pointing away from fixation) did not differ and both conditions produced shorter RTs than with neutral cues (equal signs). In contrast, neurologically intact subjects showed equal RTs with divided-attention and neutral cues, whereas valid focused cues produced reduced RTs relative to neutral cues. These results indicate that most split-brains, in contrast to normal observers, are capable of directing their attention to left and right visual field locations simultaneously, and therefore that each cerebral hemisphere controls its own visuo-spatial attention mechanism. PMID- 10678695 TI - Material-specific and non-specific attention deficits in children and adolescents following temporal-lobe surgery. AB - Attentional control in children and adolescents with unilateral temporal-lobe excisions was examined using two experimental tasks, a lexical-decision and a spatial-cue task, and a standardized vigilance task. Participants with left temporal excisions took longer than controls to reorient their attention after invalid cues compared to neutral cues in the lexical task and they made more errors on all three of the tasks. Participants with right temporal excisions differed from controls in the number of errors made on the spatial task. No differences were found between the lesion and control groups on reaction-time measures of the spatial task. The results suggest that a material-specific inhibition impairment, as well as a more general sustained attention deficit, may result after a left temporal excision in childhood or adolescence. These deficits are considered in the context of neuroanatomical models of attentional control. PMID- 10678696 TI - Interhemispheric asymmetry of the human motor cortex related to handedness and gender. AB - Most people are right-handed, preferring the right hand for skilled as well as unskilled activities, but a notable proportion are mixed-handed, preferring to use the right hand for some actions and the left hand for others. Assuming a structural/functional correlation in the motor system we tested whether asymmetries in hand performance in consistent right and left handers as well as in mixed handers are associated with anatomical asymmetries in the motor cortex. In vivo MR morphometry was used for analyzing interhemispheric asymmetry in the depth of the central sulcus in the region of cortical hand representation of 103 healthy subjects. Subjects were tested both for hand preference and hand performance. As expected, left-right differences in hand performance differed significantly between consistent right, consistent left and mixed handers and were independent on gender. Male consistent right handers showed a significant deeper central sulcus on the left hemisphere than on the right. Anatomical asymmetries decreased significantly from male consistent right over mixed to consistent left handers. Sixty two per cent of consistent left handers revealed a deeper central sulcus on the right than on the left hemisphere, but for the group as a whole this rightward asymmetry was not significant. No interhemispheric asymmetry was found in females. Thus, anatomical asymmetry was associated with handedness only in males, but not in females, suggesting sex differences in the cortical organization of hand movements. PMID- 10678697 TI - Insights from semantic dementia on the relationship between episodic and semantic memory. AB - An influential theory of long-term memory, in which new episodic learning is dependent upon the integrity of semantic memory, predicts that a double dissociation between episodic and semantic memory is not possible in new learning. Contrary to this view, we found, in two separate experiments, that patients with impaired semantic memory showed relatively preserved performance on tests of recognition memory if the stimuli were perceptually identical between learning and test. A significant effect of semantic memory was only seen when a perceptual manipulation was introduced in the episodic task. To account for these findings, we propose a revision to current models of long-term memory, in which sensory/perceptual information and semantic memory work in concert to support new learning. PMID- 10678698 TI - The calculating brain: an fMRI study. AB - To explore brain areas involved in basic numerical computation, functional magnetic imaging (fMRI) scanning was performed on college students during performance of three tasks; simple arithmetic, numerical magnitude judgment, and a perceptual-motor control task. For the arithmetic relative to the other tasks, results for all eight subjects revealed bilateral activation in Brodmann's area 44, in dorsolateral prefrontal cortex (areas 9 and 10), in inferior and superior parietal areas, and in lingual and fusiform gyri. Activation was stronger on the left for all subjects, but only at Brodmann's area 44 and the parietal cortices. No activation was observed in the arithmetic task in several other areas previously implicated for arithmetic, including the angular and supramarginal gyri and the basal ganglia. In fact, angular and supramarginal gyri were significantly deactivated by the verification task relative to both the magnitude judgment and control tasks for every subject. Areas activated by the magnitude task relative to the control were more variable, but in five subjects included bilateral inferior parietal cortex. These results confirm some existing hypotheses regarding the neural basis of numerical processes, invite revision of others, and suggest productive lines for future investigation. PMID- 10678699 TI - Large plaque parapsoriasis: clinical and genotypic correlations. AB - Twelve patients with large plaque parapsoriasis (LPP) were investigated for the presence of predominant T-cell clones, analyzing the T-cell receptor (TCR) gamma chain gene. The diagnostic and prognostic significance of TCR gene rearrangement status was assessed by a correlation with the long-term clinical follow-up. Six out of 12 patients showed a clonal T-cell population. Clinically, among the patients with clonal disease one developed clearcut mycosis fungoides (MF) after a follow-up of 8 years, in the other 5 patients no such diagnosis could be made after follow-up of 2-21 years (median: 9 years). In patients with polyclonal infiltrates the lesions remained virtually unchanged. These findings indicate that in LPP TCR gene rearrangement status has no prognostic significance and does not allow distinction of LPP and early MF. Both conditions show a clonal T-cell infiltrate with similar frequency, are very similar in clinical and histologic presentation and according to recent studies share the same low risk to develop overt MF. Therefore both terms refer to the identical clinical situation. This should be designated as early MF and efforts should concentrate on identifying those patients that are at risk to develop aggressive disease. PMID- 10678700 TI - The microanatomy of the distal arrector pili: possible role for alpha1beta1 and alpha5beta1 integrins in mediating cell-cell adhesion and anchorage to the extracellular matrix. AB - The arrector pili (AP) muscle is a small band of smooth muscle that attaches proximally to the bulge area of the pilosebaceous apparatus in the reticular dermis and extends up toward the epidermis. The distal anatomy of the AP and the anchorage mechanism allowing hair erection have not been previously described. Integrins are likely candidates mediating this attachment. Immunohistochemical techniques were used to determine the distribution of the following integrins: alpha1, alpha2, alpha3, alpha4, alpha5, alpha6 and beta1 as well as fibronectin. Frozen human scalp tissue was sectioned in traditional planes, obliquely and horizontally to visualize microanatomy in three dimensions. Histological examination revealed that the distal portions of smooth muscle fibers splay extensively between collagen bundles of the upper dermis. Integrin subunits alpha1, alpha5 and beta1 were expressed by the AP muscle. Analysis of the relative density of immunoreactivity in digitized sections revealed increased alpha5 subunit expression at the extracellular matrix (ECM)-muscle interface. These data suggest that anchorage of the AP muscle to the ECM is via alpha5beta1 integrin and alpha1beta1 integrin functions in muscle cell-cell adhesion. Extensive splaying of smooth muscle fibers may allow increased surface area contact between the ECM and smooth muscle cells expressing peripherally situated alpha5 integrin. PMID- 10678701 TI - Melanocytes in nevi and melanomas synthesize basement membrane and basement membrane-like material. An immunohistochemical and electron microscopic study including immunoelectron microscopy. AB - Light microscopic studies have shown that nevus cell nests and melanoma nests are surrounded by basement membrane (BM) material containing type IV collagen and laminin. This study confirms this by electron microscopy and relates it to proteins which interact with the basement membrane. Nevi except for dysplastic and Spitz nevi, malignant melanomas, and melanoma metastases were studied by immunohistopathology, routine electron microscopy (EM), and immunoelectron microscopy. The lesions were incubated with monoclonal antibody (moAb) against type IV collagen, laminin, and the integrin alpha6 and studied by light microscopy. In addition, melanomas were studied by immuno-EM after incubation with a moAb against matrix metalloproteinase-2 (MMP-2). Nevus cell nests and melanoma nests are surrounded by BM material containing type IV collagen and laminin by immuno-EM. The BM material various in thickness and is amorphous. Type IV collagen, laminin, and MMP-2 are synthesized by melanoma cells as well as adjacent fibroblasts. Destruction or loss of the BM is not mandatory for melanoma invasion or even metastasis. Possibly the BM material is a protective wall for melanoma cells. Interactions between melanocytes and the extracellular matrix of which the BM is a part, can be traced back to the migration of melanocytes from the neural crest. PMID- 10678702 TI - Benign mucinous metaplasia of the penis. A lesion resembling extramammary Paget's disease. AB - Benign mucinous metaplasia in the surface epithelium of the genital area is rare and has only been reported once in the vulva. A unique case of benign mucinous metaplasia of the prepuce in a 65-year-old man is reported here. The lesion measured 0.6 cm, was located in the mucous surface of the foreskin, and showed acid mucin containing cells. We regard benign mucinous metaplasia as a reactive rather than a neoplastic process. The main lesions to be considered in the differential diagnosis are mucinous syringometaplasia, extramammary Paget's disease, cutaneous squamous cell carcinoma in situ with mucinous metaplasia, superficial spreading malignant melanoma, and epidermotropic metastasis. The confinement of mucin-containing cells to the epidermis, the absence of nuclear atypia, the basal orientation of the nuclei, the predominant location of the cells in the upper layers of the epithelium, and the fact that the mucinous cells are replacing the squamous epithelium rather that infiltrating it, all assist in recognizing mucinous metaplasia of the penis as a specific and benign entity. PMID- 10678703 TI - Extranodal peripheral T-cell lymphoma with angiocentric growth pattern and Epstein-Barr viral DNA associated affecting paratesticular soft tissue. AB - Peripheral T-cell lymphomas are uncommon, accounting for only 10% to 15% of all non-Hodgkin lymphomas and their classification has been controversial. We report a case of peripheral T-cell lymphoma with angiocentric growth pattern which presented as a paratesticular tumoral nodule in a 47-year-old-man. Formalin-fixed paraffin-embedded samples from the paratesticular tumor and non-infiltrated adjacent tissue were submitted to histological, immunohistochemical, polymerase chain reaction (PCR)-based and in situ hybridization analysis. Histopathologically, there was a lymphomatous infiltrate in the paratesticular soft tissue, composed of a variable mixture of medium-sized to large cells with large cytoplasm and irregular-shaped nuclei, together with blood vessel destruction, necrosis and karyorrhexis. Immunohistochemical study revealed a high p53 expression in neoplastic cells that showed T cytotoxic immunophenotype, failing to express the natural killer (NK)-cell antigen CD56. A monoclonal rearrangement of the T-cell receptor (TCR) gamma gene by a PCR technique was demonstrated. Type-A Epstein-Barr Virus (EBV) DNA was detected by PCR-based analysis. A combined in situ hybridization and immunohistochemical study revealed that most cells labeled positive for EBV RNA showed immunostaining with the CD45RO antibody. Based on the above results, the case reported was classified as extranodal peripheral T-cell lymphoma with cytotoxic phenotype and EBV associated. The present case does not fit neatly into any of the specific types of peripheral T-cell lymphomas of the REAL classification, so a diagnosis of peripheral T-cell lymphoma unspecified was made. PMID- 10678704 TI - Glomeruloid hemangioma in POEMS syndrome shows two different immunophenotypic endothelial cells. AB - The case of a Japanese woman with glomeruloid hemangioma, an initial marker for POEMS syndrome, is reported. Her cutaneous lesions were multiple and consisted of glomeruloid hemangiomas, cherry-type capillary hemangiomas, and a mixture of both. The specimens of glomeruloid hemangiomas were studied by paraffin section immunohistochemistry with a large panel of antibodies and electron microscopy, respectively. The lesions, whose size ranged from minute foci to large nodules, were composed of anastomosing vascular channels resembling renal glomeruli and had irregular lumina, often featuring capillaries and sinusoid-like spaces. The vascular channels were lined by a single layer of endothelial cells, which showed two types of cells. The capillary-type endothelium possessed large vesicular nuclei with open chromatin and large amount of cytoplasm. The sinusoidal endothelium possessed small basal nuclei with dense chromatin as well as scant amount of cytoplasm. The former cells had a characteristic CD31+/CD34+/UEA I+/CD68- phenotype. Some of these cells ultrastructurally showed intracytoplasmic lumen formation. The latter cells had a characteristic CD31+/CD34-/UEA I-/CD68+ phenotype. The present study shows that glomeruloid hemangioma has unique morphologic and immunologic features that differ from the traditional hemangiomas as well as littoral cell angioma of the spleen. PMID- 10678705 TI - Pigmented squamous cell carcinoma. AB - Pigmented squamous cell carcinomas have been reported in the oral and ocular mucosae, but rarely in the skin. We present a case of pigmented squamous cell carcinoma of the forehead and review the current English literature. Pigmented squamous cell carcinoma can be confused with pigmented basal cell carcinomas and melanoma, especially those melanomas associated with pseudoepitheliomatous hyperplasia and should be included in the differential diagnosis of atypical pigmented lesions. PMID- 10678706 TI - Generalized pruritic eruption with suprabasal acantholysis preceeding the development of bullous pemphigoid. AB - We report a patient who presented with a papular pruritic eruption of a 3-month duration that histologically showed suprabasal acantholysis accompanied of an eosinophilic inflammatory infiltrate that was consistent with the diagnosis of Grover's disease. Later, erythematous plaques and vesicles appeared which showed a histopathological pattern of eosinophilic spongiosis. The direct immunofluorescence (DIF) study showed lineal IgG and C'3 at the dermal epidermal junction which was consistent with the diagnosis of bullous pemphigoid. No anti intercellular deposits of immunoglobulin G (IgG) or C'3 were observed. We consider that suprabasal acantholysis may represent the early phase of bullous pemphigoid. PMID- 10678707 TI - Pleomorphic giant cells in basal cell carcinoma. PMID- 10678708 TI - Cell death in pilomatricoma. PMID- 10678709 TI - Myofibroblasts and matrix components in healing palatal wounds in the rat. AB - In order to identify wound contraction and scar formation during palatal mucoperiosteal wound healing in growing rats, the temporal and spatial distribution of myofibroblasts and matrix components were determined immunohistochemically. Myofibroblasts were found in the mucosal part of the palatal wound tissue between 4 and 22 days, with the highest density at 8 days post-wounding. The number of collagen type I and type III fibers gradually increased until about 8 days postwounding, and thereafter the staining intensity of collagen type III decreased. At 60 days post-wounding there were more transversely oriented collagen type I fibers and less type III fibers and elastin present in the submucosa than in normal tissue. The results suggest that in this model wound contraction mainly takes place in the mucosa between 4 and 22 days postwounding. Furthermore, palatal wounds made in young rats heal with distinct scar tissue formation. Therefore, this model is useful to test the effects of therapies that aim to reduce wound contraction and scarring after cleft palate surgery. PMID- 10678710 TI - Synergistic enhancement of collagenous protein synthesis by human gingival fibroblasts exposed to nifedipine and interleukin-1-beta in vitro. AB - Gingival overgrowth commonly occurs coincident to therapy with calcium channel blockers. The biologic mechanism for this condition is unknown; however, many clinicians suggest that poor oral hygiene may contribute to development of the overgrowth. This study tests the hypothesis that collagenous protein synthesis by gingival fibroblasts is synergistically enhanced when they are exposed to both nifedipine (N) and the pro-inflammatory cytokine, interleukin-1-beta, a cytokine expressed in inflamed gingiva. Human gingival fibroblasts were isolated from biopsies of normal gingiva and cells separated into two groups. Group 1 was exposed to media containing 0, 5, 50, or 500 pg/ml IL-1-beta, or 10(-7) M N for 7 days; Group 2 was exposed to those concentrations of IL-1-beta +10(-7) M N. [3H] proline was added to the medium for the final 24 h. Cells and matrix were harvested and radioactivity determined by liquid scintillation analysis. Means (d.p.m./10(3) cells) were compared by factorial ANOVA and Scheffe comparisons. Collagenous protein synthesis was significantly reduced by 5 pg/ml IL-1-beta +10( 7) M N and enhanced by 500 pg/ml IL-1-beta +10(-7) M N as compared to N or IL-1 beta alone. Thus, patients may be more susceptible to gingival overgrowth coincident to nifedipine therapy as a result of the synergistic enhancement of connective tissue synthesis by these agents. PMID- 10678711 TI - Juvenile ossifying fibroma. An analysis of eight cases and a comparison with other fibro-osseous lesions. AB - Juvenile ossifying fibroma (JOF) is a well-defined clinical and histological entity that has recently been separated from other fibro-osseous lesions, including cemento-ossifying fibromas. Its biological behaviour is well defined, but unexplained. Its behaviour, clinical and histological appearance, however, bears resemblance to osteofibrous dysplasia of long bones, a lesion that in some cases has been reported to be part of a spectrum of diseases associated with adamantinoma, thus accounting for its variable biological behaviour. Eight cases of JOF were examined for islands of epithelium or single epithelial cells using immunocytochemistry. While these cases of JOF could clearly be separated from other fibro-osseous lesions, and were histologically similar to osteofibrous dysplasia, the absence of cytokeratin-positive cells in all cases suggests that another reason for its biological behaviour has still to be found. PMID- 10678712 TI - Immunolocalization of tumor necrosis factor-alpha expressing cells in recurrent aphthous ulcer lesions (RAU). AB - Tumor necrosis factor (TNF)-alpha is a pro-inflammatory cytokine and crucial mediator in many aspects of immunity. Although several studies have shown that recurrent aphthous ulcers (RAU) can be prevented by treatment that prevents the synthesis of endogenous TNF-alpha little is known about the location and distribution of TNF-alpha-expressing cells at disease sites. The aim of the present work is, therefore, to investigate TNF-alpha and its cellular distribution in RAU lesions compared with those in induced oral traumatic ulcers (TUs). Twelve biopsies of RAU lesions of oral mucosa were obtained from 12 patients with RAU. They were compared to a control group consisting of ten samples of induced TUs. All samples were analyzed for TNF-alpha expression by using monoclonal mouse anti-human TNF-alpha antibody in avidin-biotin-peroxidase complex (ABC) staining. Results were quantified by a semi-automatic VIDAS image analysis system. TNF-alpha immunoreactivity was contained mainly in monocyte/macrophages and lymphocytes within the mononuclear inflammatory infiltrates. TNF-alpha was often seen in mast cells and vascular endothelial cells in connective tissue lateral to the inflammatory infiltrates. Interestingly, 32%-60% of the mononuclear cells were found to be TNF-alpha immunoreactive in RAU lesions. TNF-alpha containing cells were more numerous in aphthae (188+/-46 cells/0.2 mm2) compared with controls (52+/-14 cells/0.2 mm2, P<0.001). These findings suggest that RAU lesions are characterized by high expression of TNF-alpha. Because such expression occurred in the mononuclear inflammatory cells, mast cells and vascular endothelial cells, TNF-alpha, which is a major inflammatory mediator, may contribute to the activation and recruitment of leukocytes that are found in RAU lesions. PMID- 10678713 TI - Lack of association between Streptococcus oralis and recurrent aphthous stomatitis. AB - In the present study, the potential involvement of Streptococcus oralis in the aetiology of recurrent aphthous stomatitis (RAS) was investigated using the polymerase chain reaction (PCR). Biopsies from 28 RAS patients were analysed, in addition to 20 oral lichen planus (OLP) and 13 normal biopsies that were used as controls. PCR was carried out using a primer pair that targets the D-alanine:D alanine ligase gene and detects DNA from both S. oralis and the closely related species Streptococcus mitis. Discrimination between these two species was achieved by digestion of PCR products with the restriction endonucleases HaeIII and HindIII, which both give distinct restriction profiles for each species. S. oralis DNA was detected in 8 of 28 (29%) RAS samples, 10 of 20 (50%) OLP samples and 6 of 13 (46%) normal samples. These results suggest that S. oralis is not of primary aetiological significance in RAS. PMID- 10678714 TI - Argyrophilic nucleolar organizer regions (AgNORs) in mucosal epithelium under experimental denture bases in rats. AB - The purpose of this study was to evaluate changes in the argyrophilic nucleolar organizer region (AgNOR) counts in mucosal epithelium induced by continuous or intermittent compressive pressure exerted through experimental denture bases and to examine the relationships between the AgNOR count, histopathological changes and the intensity of the pressure under denture bases. Continuous or intermittent compressive pressure exerted through the denture bases was applied to the hard palate of the molar region in rats. A morphometric analysis of AgNORs was performed in denture-supporting tissue 3 days and 1, 2, 4, 8, 12 and 20 weeks after the denture insertion. From the results of this study, it was found that non-pressure contact of the denture bases with palatal tissues did not change the AgNOR count. The AgNOR count was decreased by continuous or intermittent compressive pressure, and then recovered to almost the same level as with the non pressure contact at 20 weeks following a decrease of the pressure. The AgNOR counts in the epithelium under the denture bases were revealed to be related to the histopathological changes in the denture-supporting tissues and the intensity of the pressure under the denture bases. PMID- 10678715 TI - Hepatitis C virus-associated oral lichen planus: no influence from hepatitis G virus co-infection. AB - There is a variable geographic distribution in the prevalence of hepatitis C virus (HCV)-related oral lichen planus (OLP), which appears unrelated to either HCV genotype or HCV epidemiology. The present study investigated whether hepatitis G virus (HGV) co-infection may be a feature of patients with HCV related OLP, which might explain these phenomena. HGV co-infection was detected in 6 of 39 Italian patients with HCV-related OLP, but the presence of HGV did not influence the clinical presentation of OLP. It is concluded that HGV co-infection is unlikely to influence the clinical detection of HCV-related OLP. PMID- 10678716 TI - Clear cell odontogenic tumor in the mandible: report of a case with duct-like appearances and dentinoid induction. AB - A case of clear-cell odontogenic tumor with unusual histological features is presented. A 61-year-old Japanese man was admitted because of swelling of the left premolar-molar region of the mandible. Radiological examination revealed a multilocular radiolucency with irregular margins. Histological examination of the resected specimen showed infiltrative proliferation of both clear and eosinophilic cells into the adjacent soft tissue without encapsulation, suggesting the malignant potential of the tumor. The tumor cells sporadically formed cystic lesions. In addition, several tumor cell nests showed duct-like characteristics, and many eosinophilic dentin-like structures were attached to the tumor cell nests, suggesting the potential for epithelial-mesenchymal induction. Histochemically, the clear tumor cells possessed cytoplasmic glycogen granules. Both clear and eosinophilic tumor cells showed positive immunoreactivities for cytokeratin 19, epithelial membrane antigen and filaggrin, indicating an odontogenic epithelial origin. PMID- 10678717 TI - Food allergy: when and how to perform oral food challenges. AB - In many situations, the diagnosis of food allergy rests simply upon a history of an acute onset of typical symptoms, such as hives and wheezing, following the isolated ingestion of a suspected food, with confirmatory laboratory studies of positive prick skin tests or Radioallergosorbent tests. However, the diagnosis is more complicated when multiple foods are implicated or when chronic diseases, such as asthma or atopic dermatitis, are evaluated. The diagnosis of food allergy and identification of the particular foods responsible is also more difficult when reactions are not mediated by IgE antibody, as is the case with a number of gastrointestinal food allergies. In these latter circumstances, well-devised elimination diets followed by physician-supervised oral food challenges are critical in the identification and proper treatment of these disorders. Because childhood food allergies to common allergenic foods such as milk, egg, wheat and soy are usually outgrown, oral food challenges are also an integral part of the long-term management of these children. PMID- 10678718 TI - Different serum interleukin-12 and sCD30 levels in food- and pollen-sensitized children. AB - It has been proposed that a down-regulation of interleukin (IL)-12 and interferon (IFN)-gamma might be related to susceptibility to allergy in early life. The aim of this study was to assess serum IL-12 levels in food-sensitized and pollen sensitized children and to compare these with another activation marker, sCD30. Twenty children with pollen allergy and 22 food-sensitized children were included. The diagnosis of immunoglobulin (Ig)-E-mediated allergy, suggested by clinical symptoms, was based on skin-prick tests, serum IgE antibodies and total IgE levels. Samples from 24 non-allergic children were used as controls. IL-12 and sCD30 levels were measured by ELISA. It was found that pollen-sensitized patients had normal IL-12 and higher sCD30 levels than controls (114 vs. 63 U/ml, p = 0.028), but, surprisingly, food-sensitized infants showed normal sCD30 and increased serum IL-12 levels (323 vs. 118 pg/ml, p = 0.0001). No differences were found in patients suffering from asthma or allergic dermatitis. Levels of sCD30 and IL-12 determined in May showed a strong correlation with those obtained in November. Interleukin-12 and IgE levels had an inverse correlation (r = -0.494, p = 0.0001) whereas no correlation was found between sCD30 and IgE. Age had a strong negative influence on IL-12 levels in allergic (Z = 4.834, p < 0.0005) and in normal children (Z = 3.00, p < 0.002); by contrast, sCD30 levels were not significantly age-dependent. When IL-12 levels from the food-allergy group were compared with those from normal controls younger than 4 years of age, the difference remained significant (p = 0.001), ruling out an age-bias. The conclusions made in this study were that serum IL-12 and sCD30 showed different behaviors in children with food or pollen allergy. We found IL-12 and sCD30 levels in pollen-allergic patients that agree with the classical T-helper (Th) 1/Th2 paradigm of allergy. In contrast, serum IL-12 levels were increased in food sensitized children, suggesting a different immunologic pathogenesis. PMID- 10678719 TI - Infants colonized with enterotoxin-producing staphylococci at 3 months display a decreased frequency of interferon-gamma-producing CD45RO lymphocytes upon stimulation with staphylococcal enterotoxin A at birth but not at 6 months of age. AB - The aim of the study was to elucidate the relationship between the cytokine response to staphylococcal enterotoxin A (SEA) at birth and subsequent staphylococcal colonization in the first months of life. In a cohort of 45 newborns, cord blood lymphocytes were stimulated with SEA (10 ng/ml) in vitro, re stimulated with PMA (phorbol myristate acetate) and ionomycin at day 3 and assessed for CD45RO expression and cytokine generation by flow cytometry. The infants were classified into three groups according to nasal staphylococcal colonization and enterotoxin generation at 3 months: There were 16 infants with either no colonization or non-enterotoxin-producing staphylococci, 16 infants with enterotoxins B, C, D and E, and 13 infants colonized with SEA-producing staphylococci. At birth, the group without subsequent colonization displayed a significantly higher frequency of CD45RO-positive interferon-gamma-producing cells (1.7%; range 0.0-9.3%) in comparison to the SEA-positive group (0.1%; range 0.0-0.4%) and also to the group positive for other enterotoxins (0.50%; range 0.0 2.5%). Comparable but less pronounced results were found for interleukin-5 but not for interleukins 2 and 4. At 6 months, no differences in cytokine generation were detected between the three groups. The results provide evidence that a non specific immunologic immaturity at birth is a risk factor for early bacterial colonization. Furthermore, it is remarkable that this immaturity is similar to that seen in infants destined to be atopic with respect to disequilibrium of interferon-gamma to interleukin-4 generation. Thus the link between early staphylococcal colonization and subsequent atopy requires further investigation. PMID- 10678720 TI - Mononuclear cell reactivity to food allergens in neonates, children and adults. AB - A model of antigen-specific T-cell proliferative responses based on reciprocal patterns of responses to dietary and inhalant allergens has been suggested, the former being frequent in infancy but rare in adults, whereas the latter are preserved and expand between infancy and adulthood. We have evaluated the age related variations of mononuclear cell reactivity to food allergens. The cord blood mononuclear cells (CBMC) of 30 neonates without family history of atopy and the peripheral blood mononuclear cells (PBMC) of 20 healthy children and of 40 healthy adults were stimulated in vitro with beta-lactoglobulin (BLG) or ovalbumin (OVA) and the cultures were harvested after 7 days. Neonates, children and adults were compared for the percentages of positive responses and for the magnitude of response. Adult subjects showed significantly lower percentages of positive responses and reduced magnitude of response than those observed in neonates and children either in BLG or in OVA cultures. We have not observed a decrease of food allergen mononuclear cell reactivity between neonates and children for the frequency of positive responses. The magnitude of response of neonates was significantly lower than that of children in BLG cultures. Our results seem to confirm the loss of mononuclear cell reactivity to food allergens in adult age. However, other reports show conflicting data. We suggest that a rigorous standardization of the methodological steps of in vitro mononuclear cell stimulation with allergen is necessary. PMID- 10678721 TI - Cutaneous lymphocyte-associated antigen expression in children with atopic dermatitis and non-atopic healthy children. AB - Cutaneous lymphocyte-associated antigen (CLA) is a cell surface glycoprotein which has been implicated in the homing of lymphocytes to cutaneous sites. It is postulated to play an important role in T-cell migration to skin in atopic dermatitis; however, the expression of CLA in both normal children and children with atopic dermatitis has not been extensively studied. If CLA expression on T cells were important in the traffic of lymphocytes to atopic dermatitis skin lesions, it might be expected that the proportion of CLA+ T cells in unstimulated peripheral blood from children with atopic dermatitis would be elevated. We have examined the proportion of CLA+ T cells in children with atopic dermatitis and non-atopic age-matched controls. The proportion of CLA+ T cells in non-atopic children was highly associated with and increased with increasing age (r = 0.88, p < 0.001). There was no difference between the proportion of T cells expressing CLA in the unstimulated peripheral blood mononuclear cells from children with severe (p = 0.18) or with mild/moderate (p = 0.3) atopic dermatitis and age matched non-atopic controls. Despite this, children with atopic dermatitis did show evidence of perturbation of CLA expression, as unlike the non-atopic children the proportion of CLA+ T cells in the atopic children did not correlate with age. These findings suggest that while CLA expression may play a role in atopic dermatitis, other as yet undefined surface markers are likely to principally determine the migration of T cells to skin in atopic dermatitis. PMID- 10678722 TI - Dust mite allergens are carried on not only large particles. AB - The major obstacle for the successful measurement of airborne mite allergen is its very low concentration in the absence of vigorous disturbance. The aim of this study was to investigate the particle size distribution of group 2 dust mite allergen using an amplified ELISA system. Air sampling was performed using an Andersen sampler placed in the centre of the room, 1.2 m above floor level (airflow rate 28.7 l/min). This is a multistage, multiorifice cascade impactor that is comprised of six stages. Any particle greater that 4.7 microm should impact on stages 1 and 2, whilst stages 3-6 measure the predominantly respiratory range. The sampling was carried out for 30 min after 15 min of vigorous disturbance (vacuum cleaning without bag and filter). Der p 2 was measured using mAb-based ELISA with the AmpliQ amplification kit (Dako Ltd, Cambridgeshire, UK). The sensitivity was increased 15-fold as compared with standard assay, bringing the level of detection to 300 pg/ml. The majority of airborne Der p 2 (79.4%) was carried on large particles (> 4.7 microm). However, a small but important proportion of airborne Der p 2 (20.6%) was associated with small particles (1.1 4.7 microm). It is worth noting that all the levels measured were below the detection limit of standard assay. In conclusion, we have shown that using an amplification system, airborne mite allergen previously undetectable owing to its low concentration can be quantified. Group 2 dust mite allergen is carried not only on large particles. A small, but potentially significant proportion of this airborne allergen is associated with small particles which, when inhaled, may penetrate deep into the human respiratory tract. PMID- 10678723 TI - Prevalence of asthma and allergic diseases in primary school children in Ankara, Turkey: two cross-sectional studies, five years apart. AB - The prevalence of allergic diseases is reported to have increased worldwide. Two questionnaire surveys, five years apart, were conducted to evaluate the trend of prevalence rates and possible risk factors among primary school children in Ankara, Turkey. A previous survey in 1992 revealed the lifetime prevalences of asthma, wheezing, allergic rhinitis and atopic dermatitis were 17.4%, 23.3%, 28% and 6.1%, and the prevalences for the last 12 months were 8.3%, 11.9%, 15.4% and 4%, respectively. The survey was repeated with the same questionnaire in the same age group (6-13 years) of the same school in May 1997. The parents of 358 boys and 380 girls completed the questionnaire. The lifetime and last 12 months' prevalences of asthma, wheezing, rhinitis and atopic dermatitis were 16.8%, 22.5%, 18.7%, 6.5%, and 9.8%, 13.3%, 14.1%, 4.3%, respectively. There was a significant change only for the lifetime prevalence of rhinitis (p < 0.001). The rate of indoor smoking had declined from 73.9% to 64%, and pet ownership had risen from 7.9% to 22.9% (p < 0.001 for both). Atopic family history was the most prominent risk factor for all types of allergic disorders. Male gender was a significant risk factor for current asthma and wheezing [odds ratio (OR) = 1.80 and 1.59; 95% confidence intervals (CI) = 1.09-2.98 and 1.01-2.48, respectively], and passive smoking affected the occurrence of allergic rhinitis (OR = 1.84; CI = 1.13-3.00). The prevalence rates of allergic diseases among primary school children in Ankara stabilized during a 5-year period for all diseases other than allergic rhinitis. However, there are changing behavior patterns, i.e. indoor smoking and keeping pet animals, which that may have affected these rates. PMID- 10678724 TI - Use of pocket-sized turbine spirometer in monitoring exercise-induced bronchospasm and bronchodilator responses in children. AB - For field studies of asthma, portable hand-held pulmonary function testing devices are required. Other than for peak flow measurements, little has been done to validate their use in children. Fifty children aged 5-15 years having asthma symptoms were examined using an exercise challenge (8 min free running outdoors) and a bronchodilation test (salbutamol inhalation at a dose of 0.15 mg/kg). Pulmonary function was measured with a turbine spirometer, with a Wright peak flow meter (WPEF) and with a flow-volume spirometer (FVS). A fall of 15% or more in peak expiratory flow associated with wheezing or cough was considered diagnostic for bronchial hyper-responsiveness to exercise (BHRE). A rise of 15% or more from baseline in peak expiratory flow after salbutamol inhalation was considered as a positive bronchodilator response (BDR). BHRE was present in 16 children (32%). Using the limit of a 15% or greater fall in FEV1, turbine spirometry identified 12 as BHRE-positive and no additional cases, giving a sensitivity of 75% and a specificity of 100%. The turbine spirometer showed lower FEV1 values than the FVS, the difference increasing with airway obstruction. BDR was positive in eight children (16%). Using the limit of a 10% or greater rise in FEV1, turbine spirometry was positive in six cases. FEV1 measured by turbine spirometry could not be used interchangeably with conventional FVS. However, the turbine spirometer offers the possibility to measure FEV1 repeatedly in field conditions, such as during exercise challenges outdoors. PMID- 10678725 TI - Allergy to an occupational allergen (Sapelli wood) in a child. AB - The case is presented of a child who developed rhinoconjunctivitis, angioedema and asthma by sensitization to Sapelli wood, which was used in his father's carpentry. Positive skin-prick test, high levels of specific immunoglobulin E by ELISA and a positive conjunctival challenge test suggest a type I hypersensitivity mechanism to this wood. This is the first case report of sensitization to Sapelli wood and it confirms that occupational allergen exposure as Sapelli wood may also cause sensitization in a child. PMID- 10678726 TI - The clinical interpretation of skin prick tests (SPT) PMID- 10678727 TI - Anatomical bases for paravertebral anesthetic block: fluid communication between the thoracic and lumbar paravertebral regions. AB - An injection of a local anesthetics in the paravertebral region produces an analgesic field on the same side of the body, a paravertebral block. One point in question about this block is whether the local anesthetic spreads from the thoracic to the lumbar level of the paravertebral region. The purpose of this study was to find how the anesthetic fluid traveled to the lumbar paravertebral region, if at all. Twelve cadavers were used in this study. 15 ml of crimson dye was injected into the paravertebral region at the 11th thoracic level. The viscerae were removed so that we could examine the dye spread. While the crimson dye spread in the endothoracic fascia posterior to the parietal pleura, it also spread downward in the fascia mostly along the splanchnic nerves. At the upper surface of the diaphragm the dye spread laterally in the fascia, and entered the abdominal cavity through the medial and lateral arcuate ligaments. In the abdominal cavity, the dye was found to have spread so widely in the transversalis fascia that the subcostal, iliohypogastric, ilioinguinal, genitofemoral, lateral femoral cutaneous and femoral nerves were involved. We concluded that the dye in the thoracic paravertebral region can enter the abdominal cavity through the medial and lateral arcuate ligaments. This study explained possible fluid communication between the thoracic and lumbar paravertebral regions and confirmed our former clinical observations. The result is important for the future clinical application of paravertebral anesthesia. PMID- 10678728 TI - High and low lateral approaches to the popliteal artery. AB - Using a series of 20 dissections and two anatomic transverse sections of a lower limb, the authors investigated the lateral approaches to the popliteal artery. The high lateral approach (above the knee) is not very aggressive and gives access to the retro-genicular part of the popliteal artery. After cutaneous and fascial incision, a simple gap between the vastus lateralis and biceps femoris mm. allows easy exposure of the popliteal vessels after backward retraction of the sciatic nerve. The low lateral approach to the artery (below the knee) is very aggressive for the vessels, nerves, and ligaments of the area. It involves the resection of the upper fourth of the fibula and the isolation and protection of the common peroneal nerve Nevertheless, these lateral approaches must be known and used when classic approaches (medial and posterior) are impossible. PMID- 10678729 TI - Topoanatomic relations of the ophthalmic artery viewed in four horizontal layers. AB - In the present paper we have studied the gross anatomy of the ophthalmic artery in 200 human cadaver dissections viewed in four horizontal layers. The ophthalmic artery can be divided into the origin, intracranial, intracanalicular and intraorbital parts. The most common origin of the artery was from the medial half of the anterior side of the internal carotid artery (ICA) in its upper curve (52%), followed by the medial half of the superior side (44%), and in only 4% from the medial side of the upper curve of the ICA. We have examined the topographic anatomy of the ophthalmic artery in detail, and found a rare anastomosis of the ophthalmic artery with the frontal branch of the middle meningeal artery. PMID- 10678730 TI - Biomechanical behaviour in vitro of the spine and lumbosacral junction. AB - Six fresh human specimens extending from the 9th thoracic vertebra (T9) to the pelvis were used to study the biomechanical behaviour of the long lumbopelvic segments, including mobility of the sacrum. The loads were applied at T9 using pure couples up to 5 Nm. The displacements were measured by an optoelectronic method (VICON 140). Stress-displacement curves were obtained for the three angular components of the vertebra studied according to the plane of the principal stresses and of the two other planes corresponding to the coupled mobilities. Mobility decreased from T9 to the sacrum. There was mobility of the sacrum in relation to the pelvis in flexion, with a mean of 1.28 degrees (0.5 to 2.8 degrees); 3 sacrums showed a mobility of the order of one degree for torsional stresses. There was no sacral mobility during stresses in lateral flexion. The use of this experimental protocol with low mechanical stresses should allow the evaluation of long osteosyntheses extending to the sacrum. PMID- 10678731 TI - A new radiographic method for evaluation of the position of the carpus in the coronal plane: results in normal subjects. AB - The methods used to quantify pathological variations of the position of the carpus in the coronal plane, mainly ulnar translation of the carpus from trauma or rheumatoid disease, are often difficult to use in arthritic or postsurgical wrists; moreover, they require the measurement of the whole length of the third metacarpal. The aim of this study was to determine a reliable and easy-to-use index to analyse the position of the carpus in the coronal plane. One hundred PA X-rays of normal wrists were studied, of which 56 presented with a medial hamate facet of the lunate. An index of position of the carpus in the coronal plane is defined as the ratio of orthogonal distances from [1] the most medial point of the distal radius and [2] the most medial point of the capitate to the long axis of the radius: its mean value being 1.06 (+/- 0.13) in this series of normal wrists. The index is influenced by the presence of a medial hamate facet of the lunate, but is not dependent on the ulnar head and radial styloid process, nor on the length of the third metacarpal. To help to define the usefulness of the index in quantifying the different types of ulnar carpal translations in clinical practice, further studies are required. PMID- 10678732 TI - Importance of elliptic Fourier methods for morphometry of complex outlines: application to the distal human femur. AB - The purpose of this technical note is to present an automatic procedure of shape characterisation using new developments in elliptical Fourier methods combined with image analysis techniques. It was applied, as an example, to the outline of the distal extremity of the distal human femur in inferior axial view. This outline was automatically extracted and characterised by an ordered series of harmonics, each harmonic being described by four new parameters called elliptical descriptors. Step by step reconstructions of outlines using an increasing number of harmonics were than performed. The simultaneous study of the elliptic descriptors and of the step by step reconstructions allowed a considerably easier geometric and morphologic interpretation of the harmonic contributions than classically. The main morphologic features of the distal femur (lateral and medial condyles, intercondylar fossa, lateral and medial borders of the articular surface, and patellar groove) were mainly described by the first seven harmonics. These new developments in elliptic Fourier methods open interesting perspectives for the study of complex outlines, providing an accurate individual morphologic characterisation, and thus the possibility of polymorphic analysis. PMID- 10678733 TI - The neurovascular and muscular anomalies of the gluteal region: an atypical pudendal nerve. AB - In two cases, one male and one female, muscular anomalies together with neurovascular variations were encountered in the gluteal regions, in each cadaver on the same side. In the male cadaver, there was a double piriformis muscle and high division of the sciatic nerve. In the female cadaver, in addition to these anomalies, the superior and inferior gemelli and obturator internus muscles, and the internal pudendal vessels and pudendal nerve passed behind the sacrotuberous ligament. Although duplication of the piriformis and high division of the sciatic nerve have been reported previously, to the best of our knowledge the other anomalies have not yet been reported. The abnormal relationship of the internal pudendal vessels and the pudendal nerve with the sacrotuberal ligament, as in our case, may cause venous congestion, arterial obstruction, dysfunction of penile erection and perineal neuralgia. These anomalies of the gluteal region are not only of academic interest, but may be of practical importance for surgical intervention in the area. PMID- 10678734 TI - Double gluteus maximus muscle with associated variations in the gluteal region. AB - We report bilateral muscular and neurovascular anomalies of the gluteal region in a cadaver. On the right side, the gluteus maximus muscle had two parts, one of which was fibrous and the other muscular. In addition, there were duplicated piriformis muscles and high division of the sciatic nerve. The common peroneal nerve passed between the two parts of the piriformis muscle, and the tibial nerve emerged from under the lower piriformis muscle (infrapiriform foramen). At the same time the internal pudendal vessels and pudendal nerve passed over the sacrotuberous ligament on the left side. The double piriformis muscles and high division of the nerve are known as an anomaly which is believed to cause a nerve compression syndrome called the syndrome of the piriformis muscle. To the best of our knowledge anomalies of the gluteus maximus muscle and pudendal structures have not yet been reported. This complex anomaly should be kept in mind in connection with intramuscular injections of the gluteal region, the piriformis syndrome, and the surgery of this region. PMID- 10678735 TI - Role of hip abduction in the kinetics of the thoracic and lumbar spine. AB - The motion of the thoracic and lumbar spine segments was analysed by mean of the skin landmarks method in 14 volunteers (7 males, 7 females). The results demonstrated that there was no significant difference between men and women and there was either facilitation or limitation of the motion of the characteristic landmarks of spinal segments during trunk motion in perpendicular planes. The mobility of spinal segments was associated with pelvic girdle motion in order to obtain the planned amplitude, and varied with hip joint abduction. The analysis of the precise mechanisms of relaxing or stretching of the pelvivertebral musculo ligamentous structures constitutes a large field of investigation that necessitates the study of other lower limb postures and the use of robotic techniques. PMID- 10678736 TI - Physiological time series: distinguishing fractal noises from motions. AB - Many physiological signals appear fractal, in having self-similarity over a large range of their power spectral densities. They are analogous to one of two classes of discretely sampled pure fractal time signals, fractional Gaussian noise (fGn) or fractional Brownian motion (fBm). The fGn series are the successive differences between elements of a fBm series; they are stationary and are completely characterized by two parameters, sigma2, the variance, and H, the Hurst coefficient. Such efficient characterization of physiological signals is valuable since H defines the autocorrelation and the fractal dimension of the time series. Estimation of H from Fourier analysis is inaccurate, so more robust methods are needed. Dispersional analysis (Disp) is good for noise signals while bridge detrended scaled windowed variance analysis (bdSWV) is good for motion signals. Signals whose slopes of their power spectral densities lie near the border between fGn and fBm are difficult to classify. A new method using signal summation conversion (SSC), wherein an fGn is converted to an fBm or an fBm to a summed fBm and bdSWV then applied, greatly improves the classification and the reliability of H, the estimates of H, for the times series. Applying these methods to laser-Doppler blood cell perfusion signals obtained from the brain cortex of anesthetized rats gave H of 0.24+/-0.02 (SD, n=8) and defined the signal as a fractional Brownian motion. The implication is that the flow signal is the summation (motion) of a set of local velocities from neighboring vessels that are negatively correlated, as if induced by local resistance fluctuations. PMID- 10678737 TI - Heart fatty acid binding protein and cardiac troponin T plasma concentrations as markers for myocardial infarction after coronary artery ligation in mice. AB - Ligation of the main left coronary artery in mice serves as a model for myocardial infarction (MI). We tested whether plasma concentrations of heart-type fatty acid-binding protein (H-FABP) and/or cardiac troponin T (cTnT) discriminate between infarcted and sham-operated mice and allow estimation of infarct size. Mice were subjected to coronary artery ligation or sham surgery and release curves of H-FABP and cTnT were determined. At 4 h after surgery the mean (+/-SD) H-FABP plasma concentration was 461+/-134 microg/l (n=10) in MI and 185+/-51 microg/l (n=6; P<0.001) in sham-operated mice. By 24 h after surgery H-FABP levels had returned to normal in both groups. cTnT plasma concentrations increased up to 48 h after MI to 13.5+/-6.2 microg/l (n=6; P<0.001) compared with 0.031+/-0.063 microg/l (n=7) in sham-operated mice. Linear regression analysis revealed a significant correlation between plasma H-FABP at 4 h and infarct size assessed 7 days after surgery. Plasma cTnT did not correlate significantly with infarct size. In conclusion, plasma cTnT concentration at 48 h after infarction can be used to distinguish MI from sham mice, whereas H-FABP concentration at 4 h can be used for stratification of animals according to infarct size. PMID- 10678738 TI - Modulation of cloned skeletal muscle sodium channels by the scorpion toxins Lqh II, Lqh III, and Lqh alphaIT. AB - The scorpion alpha-toxins Lqh II, Lqh III, and Lqh alphaIT from Leiurus quinquestriatus hebraeus are representatives of typical alpha-toxins, specific for either mammals (Lqh II) or insects (Lqh alphaIT), and alpha-like toxins (Lqh III) which act on both mammals and insects. For a comparative study of the effects of these toxins on mammalian sodium channels we stably expressed rat skeletal muscle sodium channel alpha subunits (microI) in HEK 293 cells and measured Na+ currents in the whole-cell patch-clamp mode. The alpha- and alpha like toxins strongly slowed down channel inactivation with a half-maximal effect at 1.4 nM (Lqh II), 5.4 nM (Lqh III), and 0.5 nM (Lqh alphaIT). The recovery from fast inactivation was accelerated by all toxins with the potency sequence: Lqh II>Lqh alphaIT>Lqh III. The voltage dependence of inactivation and recovery from inactivation were reduced while the threshold for activation was only slightly shifted by approximately equal to 10 mV without altering the slope factors, suggesting uncoupling of the impaired inactivation from the activation. The toxins induced an increase in peak inward current, which was accounted for by an increased maximal open-channel probability. Although all three toxins induced similar modifications of the channel properties, their kinetics of association and dissociation were very different. Between -140 and -80 mV toxin association was not voltage dependent. In 100 nM toxin the association time constants were: 1.3 s (Lqh II), 20 s (Lqh III), and 3.8 s (Lqh alphaIT). At positive voltages the toxin dissociated from the channel; at +100 mV the dissociation time constants were 30, 321, and 135 ms, respectively. In contrast to the association, dissociation was voltage dependent with a similar slope of about 12 mV per e-fold change for all three toxins. The strong differences in the association and dissociation kinetics of these toxins may identify them as members of different scorpion alpha-toxin subgroups. PMID- 10678740 TI - Endogenous nitric oxide attenuates erythropoietin gene expression in vivo. AB - This study aimed to investigate the role of endogenous nitric oxide (NO) in erythropoietin (EPO) gene expression in mice in vivo. For this purpose EPO mRNA was semiquantitated by ribonuclease protection assay in livers and kidneys of three groups of mice: wild-type (wt), endothelial NO-synthase (NOS) knockout mice (eNOS-/-), and wt treated with the NOS inhibitor N(G)-nitro-L-arginine methyl ester (50 mg x kg(-1) x day(-1)) for 4 days (wt+L-NAME). EPO gene expression was stimulated by normobaric hypoxia (8% O2) or by 0.1% carbon monoxide (CO) inhalation for 4 h each, or by intraperitoneal injection of 60 mg/kg cobaltous chloride (CoCl2) for 6 h. Renal EPO mRNA in wt increased 12-, 40-, and 13-fold over normoxic levels in response to hypoxia, CO and CoCl2 respectively. EPO mRNA was detectable in the livers only after CO exposure. Renal and hepatic EPO gene expression in wt+L-NAME appeared moderately increased relative to wt with a maximal 2.5-fold enhancement after CO exposure. EPO mRNA levels in eNOS-/- mirrored those of wt+L-NAME, but the effects were less prominent. Our data suggest that endogenous NO attenuates EPO gene expression in mice. This effect is dependent on the rate of EPO gene induction. PMID- 10678741 TI - Structural and cellular adaptation of duodenal iron uptake in rats maintained on an iron-deficient diet. AB - Iron deficiency induced in rats maintained on a commercial diet with a low iron content has been used to investigate adaptive mechanisms that enhance duodenal iron uptake. These adaptive changes have been divided into those that result from changes in villus surface area (structural adaptation) and those that reflect changes in the way individual enterocytes express iron transport function (cellular adaptation). Cellular adaptation was assessed by carrying out microdensitometry of autoradiographs prepared from duodenal tissue previously incubated for 5 min in 200 micromol/l 59Fe2+-ascorbate. Structural adaptation was studied by performing image analysis of microdissected and sectioned villi. Cellular adaptation involved increased iron uptake by enterocytes present in the lower villus. Thus iron deficiency resulted in a threefold enhanced expression of uptake in the lower 100 microm villus (3.9+/-2.4 versus 12.6+/-1.5 arbitrary units, P<0.001). Maximal uptake was reached in the upper region of both control and iron-deficient villi, but iron deficiency had no effect on cellular uptake at this part of the villus. Structural adaptation involved the lengthening (+16%, P<0.05) and broadening (+14%) of villi in the duodenum of iron-deficient rats. The resultant expansion in villus area caused a further increase in uptake that was mostly expressed in the upper villus. Maximal uptake corrected for structure occurred in the middle third of villi from control and iron-deficient rats. Cellular plus structural adaptation produced a twofold increase in iron uptake. More than half of this effect was caused by changes in villus structure. [3H]Thymidine labelling experiments revealed a slightly earlier expression of enterocyte iron uptake in iron deficiency. PMID- 10678739 TI - Calcium, ATP and nuclear pore channel gating. AB - Nuclear envelope (NE) cisternal Ca2+ and cytosolic ATP are required for nuclear pore-complex-(NPC-) mediated transport of DNAs, RNAs, transcription factors and other large molecules. Isolated cardiomyocyte nuclei, capable of macromolecular transport (MMT), have intrinsic NPC ion channel behavior. The large ion conductance (gamma) activity of the NPC channel (NPCC) is blocked by the NPC monoclonal antibody mAb414, known to block MMT, and is also silenced during periods of MMT. In cardiomyocytes, neither cytosolic Ca2+ nor ATP alone directly affects NPCC gating. To test the role of Ca2+ and ATP in NPCC activity, we carried out the present patch-clamp study with the pipette attached to the outer NE membrane of nuclei isolated from cultured Dunning G prostate cancer cells. Our investigations demonstrate that in these isolated nuclei neither cytosolic Ca2+ nor ATP alone directly affects NPCC gating. However, when simultaneously applied to the bath and pipette, they transiently silence NPCC activity through stimulation of MMT by raising the Ca2+ concentration in the NE cisterna ([Ca2+]NE). Our fluorescence microscopy observations with nuclear-targeted macromolecular fluorochromes (B-phycoerythrin and plasmid for the enhanced green fluorescence protein EGFP, pEGFP-C1) and with FITC-labeled RNA support the view that channel silence accompanies MMT. Repeated Ca2+ loading of the NE with Ca2+ and ATP, after unloading with 1-5 microM inositol 1,4,5-trisphosphate (IP3), thapsigargin (TSG) or 5 mM BAPTA or EGTA, failed to affect channel gating. This result indicates that other factors are involved in this phenomenon and that they are exhausted during the first cycle of NE Ca2+ loading/unloading--in agreement with current theories of NPC-mediated MMT. The results explain how Ca2+ and IP3 waves may convert the NE into an effective Ca2+ barrier and, consequently, affect the regulation of gene activity and expression through their feedback on MMT and NPCC gating. Thus, [Ca2+]NE regulation by intracellular messengers is an effective mechanism for synchronizing gene activity and expression to the cellular rhythm. PMID- 10678742 TI - Lithium activates mammalian Na+/H+ exchangers: isoform specificity and inhibition by genistein. AB - Replacement of external NaCl with LiCl induced cytoplasmic alkalinization in CCL 39 cells and rat L6 myoblasts expressing the endogenous Na+/H+ exchanger isoform NHE1. This Li+-induced alkalinization is due to activation of the Na+/H+ exchanger because it was completely inhibited by 100 microM ethylisopropylamiloride (apparent Kd=1 microM) and because it did not occur in exchanger-deficient PS120 cells. The effect of Li+ was not mimicked by Na+, K+, Cs+ and choline+. Li+ caused cytoplasmic alkalinization in PS120 cells expressing NHE1 or NHE2, but not NHE3, when Li+ was added to cells at a concentration high enough to saturate their external transport sites as predicted from Li+ affinities. Li+ did not induce phosphatidylinositol (PI) turnover or intracellular Ca2+ mobilization. Li+-induced alkalinization was not affected by protein kinase C down-regulation, loss of glycogen synthase kinase 3beta caused by antisense oligonucleotide treatment, or pretreatment with calphostin C, pertussis toxin, MEK inhibitor PD98059 and PI3-kinase inhibitor LY294002. However, it was markedly suppressed by the tyrosine kinase inhibitor genistein (10 microM). Thus, externally added Li+ activates NHE1 and NHE2 via a mechanism possibly involving a tyrosine kinase, causing an increase in cytoplasmic pH that could potentially affect various cell functions. PMID- 10678743 TI - Developmental changes in chemoreceptor nerve activity and catecholamine secretion in rabbit carotid body: possible role of Na+ and Ca2+ currents. AB - In order to better understand the post-natal increase in peripheral chemoreceptor responsiveness to hypoxia, chemoreceptors of newborn (1-2 days) and older (10-12 days, 30 days, adult) rabbits were isolated and superfused, in vitro. The free tissue catecholamine concentration was measured using carbon-fiber voltammetry and pauci-fiber nerve activity was recorded from the sinus nerve during stimulation (4 min) with graded hypoxia or increased potassium. Both the peak catecholamine and peak nerve responses to stimulation with 10% and 0% oxygen increased with age, particularly between 10 and 30 days of age. In contrast, peak nerve and peak catecholamine responses to increased potassium did not significantly change with age. For a better understanding of how responsiveness increases with age, the fast Na+ and the Ca2+ currents were measured from isolated glomus cells of newborn and older rabbits, but the magnitude of the currents when normalized to membrane area was not significantly different between ages. We conclude that: (1) rabbit chemoreceptors mature in the newborn period (10-30 days) and part of this maturation is an increase in catecholamine secretion, (2) maturation of hypoxia transduction primarily occurs in steps prior to depolarization since potassium-evoked responses were not affected, and (3) an increase in the magnitude of glomus cell fast Na+ or Ca2+ currents is not a likely mechanism for the maturational change, but changes in the oxygen sensitivity of these currents cannot be excluded. PMID- 10678744 TI - Inner-medullary organic osmolytes and inorganic electrolytes in K depletion. AB - The renal concentrating defect typical for chronic K depletion has been ascribed to malfunction of renomedullary cells caused by inadequate accumulation of organic osmolytes. A reduction in intracellular ionic strength, which is believed to influence decisively the accumulation of organic osmolytes, has been held responsible for insufficient osmolyte accumulation. To test this hypothesis, intra- and extracellular Na, Cl and K concentrations, the major determinants of ionic strength, were measured in the papilla by electron microprobe analysis and organic osmolytes (glycerophosphorylcholine, betaine, sorbitol, myo-inositol, free amino acids) in inner-medullary tissue by HPLC in antidiuretic rats kept on either a control (normal-K) or a K-deplete (low-K) diet and in euhydrated rats with free access to water and control diet. K depletion was associated with a reduced urine concentrating ability. Papillary interstitial ionic strength (sum of Na, Cl and K) in antidiuretic low-K rats was significantly reduced compared with antidiuretic normal-K rats (688+/-19 vs. 971+/-61 mmol/kg wet wt) but was similar to that in euhydrated normal-K rats (643+/-35 mmol/kg wet wt). The lower interstitial ionic strength in antidiuretic low-K and euhydrated normal-K rats was associated with a lower total content of organic osmolytes in the inner medulla (365+/-14 and 381+/-20, respectively, vs. 465+/-11 mmol/kg protein in antidiuretic normal-K rats). Intracellular ionic strength (sum of Na, Cl and K) of papillary collecting duct cells, however, was similar in antidiuretic normal-K and euhydrated normal-K rats (171+/-5 and 179+/-11 mmol/kg wet wt) but lower in antidiuretic low-K rats (138+/-9 mmol/kg wet wt). These results do not support the view that, in the steady state of osmotic adaptation of renomedullary cells in situ, intracellular ionic strength is the decisive factor for maintaining high levels of organic osmolytes. During chronic K depletion, reduced osmolyte accumulation by renomedullary cells may be the consequence, rather than the cause, of lower medullary interstitial tonicity. PMID- 10678745 TI - Trans/paracellular, surface/crypt, and epithelial/subepithelial resistances of mammalian colonic epithelia. AB - The epithelial barrier function of the large intestine resides in the trans- and paracellular pathways of the surface epithelium and crypts. Conventional transmural resistance and permeability measurements, however, yield only the resistance of the whole tissue and not that of its individual components. Combining conductance scanning techniques and impedance analysis, we determined the resistance of epithelial and subepithelial tissues, crypts and surface epithelium, and trans- and paracellular pathways of the mouse distal colon. The subepithelial tissue contributed 15% to the transmural resistance of 118+/-9 omega x cm2. In the epithelium proper the resistance of crypts (429+/-86 omega x cm2) exceeded that of the surface epithelium (132+/-15 omega x cm2). The paracellular resistance (3.2+/-0.4 k omega x cm2) of the surface epithelium was 23-fold higher than the transcellular resistance (137+/-16 omega x cm2), and thus the epithelium was classified as "medium tight". In order to investigate the trans- and paracellular resistances of the crypt epithelium as well, flat monolayers of HT-29/B6 cultured colon crypt cells were studied, which had a transepithelial resistance of 349+/-32 omega x cm2. With transcellular resistance (377+/-41 omega x cm2) tenfold lower than the paracellular resistance (3.9+/-1.3 k omega x cm2), this cryptal monolayer was also classified as "medium tight". Hence, considering the 1.2 times larger area of the crypt epithelium, the surface epithelium has a 4 times larger ion permeability than the crypt epithelium. However, the paracellular resistances are not different. Thus the lower transcellular resistance of the surface compared to the crypt epithelium suggests a higher density of ion channels in the apical membrane of surface cells. PMID- 10678746 TI - Operation Everest III: energy and water balance. AB - We hypothesized that hypoxia decreases energy intake and increases total energy requirement and, additionally, that decreased barometric pressure increases total water requirement. Energy and water balance was studied over 31 days in a hypobaric chamber at 452-253 Torr (corresponding to 4,500-8,848 m altitude), after 7 days acclimatization at 4,350 m. Subjects were eight men, age 27+/-4 years (mean+/-SD), body mass index 22.9+/-1.5 kg/m2. Food and water intake was measured with weighed dietary records, energy expenditure and water loss with labelled water. Insensible water loss was calculated as total water loss minus urinary and faecal water loss. Energy intake at normoxia was 13.6+/-1.8 MJ/d. Energy intake decreased from 10.4+/-2.1 to 8.3+/-1.9 MJ/d (P<0.001) and energy expenditure from 13.3+/-1.6 to 12.1+/-1.8 MJ/d (P<0.001) over the first and second 15-day intervals of progressive hypoxia. Absolute insensible water loss did not change (1.67+/-0.26 and 1.66+/-0.37 l/d), however, adjusted for energy expenditure it increased with ambient pressure reduction (P<0.05). In conclusion, hypoxia induced a negative energy balance, mainly by a reduction of energy intake. Overall insensible water loss was unchanged because the increase in respiratory evaporative water loss was counterbalanced by a decrease in metabolic rate that probably limited the hypoxia-induced increase in ventilation. PMID- 10678747 TI - Chronic oxidative stress in the RVLM modulates sympathetic control of circulation in pigs. AB - Oxidative stress is a key event in the pathogenesis of several cardiovascular diseases and may be similarly induced by long-term treatment with organic nitrates. We examined the effects of inhibiting extracellular oxidative stress in the rostral ventrolateral medulla (RVLM), the brain stem area which primarily controls sympathetic tone. Superoxide dismutase (SOD, 10 U/microl) was microinjected into the RVLM of anesthetized pigs that were either untreated (control, n=10), treated for 4 weeks with the organic nitrate isosorbidedinitrate (ISDN, 4 mg kg(-1) day(-1), n=6) or ISDN-treated followed by a 2-week recovery period (recovery, n=4). In control animals SOD produced moderate inhibitory effects on baseline sympathetic activity, indicated by decreases in renal sympathetic nerve activity (RSNA), mean arterial blood pressure (MAP), and heart rate (HR) without causing changes in femoral vascular conductance (FC). These effects of SOD were greatly enhanced in ISDN-treated pigs. Following the recovery period, SOD again produced smaller effects in the RVLM but they were, however, still significantly greater than in untreated animals. In contrast, the transmission of sympathoexcitatory reflexes by the RVLM, as evoked by sciatic nerve stimulation, was not affected by SOD injections in either experimental group. Furthermore, the number of NO-synthase-positive neurons in the RVLM region was significantly reduced both in ISDN-treated and the recovery pigs, suggesting that oxidative stress caused sustained changes in NOS activity within the brain stem. These data suggest that excitatory actions of oxidative stress contribute significantly to the generation of baseline sympathetic tone in the RVLM during long-term treatment with organic nitrates. Similar mechanisms could promote sympathetic tone in cardiovascular diseases that are associated with endogenous oxidative stress for longer periods. PMID- 10678748 TI - Lysophosphatidylcholine triggers intracellular calcium release and activation of non-selective cation channels in renal arterial smooth muscle cells. AB - The effects of a lipid component of oxidized low-density lipoproteins (ox-LDL), L alpha-palmitoyl-lysophosphatidylcholine (LPC), on membrane currents of isolated canine renal artery smooth muscle cells (RASMC) were examined using the whole cell configuration of the patch-clamp technique. In RASMC exposed to nominally Ca2+-free solutions and dialyzed with 0.1 mM EGTA and 140 mM K+, superfusion with LPC (10 microM) elicited spontaneous transient outward currents (STOCs) and/or spontaneous transient inward currents (STICs), followed by the activation of a large voltage-independent current with a reversal potential (Er) close to 0 mV. Buffering intracellular Ca2+ with 10 mM BAPTA prevented the appearance of STOCs and STICs, but not the activation of the voltage-independent current. Er of the LPC-induced voltage-independent current exhibited sensitivity to changes in [K+]o and [Na+]o in a manner consistent with a non-selective cation current (I(NSC)) and was blocked by gadolinium (Gd3+; 10 microM). Shifts in Er of the LPC-induced I(NSC) in response to changes in [Ca2+]o were used to estimate a relative Ca2+ to Na+ permeability ratio (P(Ca)/P(Na)) of 1.67. These results suggest that LPC causes abnormal sarcoplasmic reticulum Ca2+ regulation, leading to the appearance of STOCs and STICs, and the activation of I(NSC) in vascular smooth muscle cells. These effects may explain the ability of ox-LDLs to elevate [Ca2+]i in vascular smooth muscle and inhibit endothelium-dependent relaxation. PMID- 10678749 TI - Quinpirole attenuates the striatal immediate early gene expression, but not the hyperactivity, induced by the serotonin agonist RU-24969. AB - Systemic administration of the mixed 5-HT(1A/1B) agonist RU-24969 has been shown to produce a dramatic increase in locomotor activity and to induce robust c-Fos expression in the rat striatum. Previous studies have also shown that pretreatment with the D2-like dopamine agonist quinpirole virtually abolishes RU 24969-induced striatal c-Fos expression. The present study was undertaken to determine whether the effects of RU-24969 on immediate early gene expression extend to the additional Fos family transcription factors FosB and Fra-2. Additionally, this study quantitatively examined the effect of quinpirole pretreatment on the ability of RU-24969 to induce both locomotor hyperactivity and striatal immediate early gene expression. RU-24969 alone produced elevations in locomotor activity and induced clear expression of c-Fos, FosB and Fra-2 throughout the entire striatal complex. Quinpirole pretreatment virtually abolished RU-24969-induced expression of all three transcription factors, but did not alter the elevated locomotor activity produced by RU-24969. These results demonstrate that the effects of RU-24969 on locomotor activity can be dissociated from its effects on immediate early gene expression within the striatum. PMID- 10678750 TI - Mapping of the progressive metabolic changes occurring during the development of hippocampal sclerosis in a model of mesial temporal lobe epilepsy. AB - We have recently characterized the histopathological changes in an experimental model of mesial temporal lobe epilepsy (MTLE) induced by the intrahippocampal injection of low dose of kainate in mice. Although cerebral metabolism and blood flow are extensively studied and used in human MTLE to locate the regions involved in seizures before surgery, this exploration is only performed once the disease has fully developed. Therefore, in the present study, we followed the temporal evolution of intrahippocampal kainate-induced metabolic changes in mice from kainate injection to 120 days later by the quantitative autoradiographic [14C]2-deoxyglucose (2DG) technique. At day 0 (late phase of status epilepticus (SE)) and 15 days after kainate, i.e., during the period of ongoing neuropathological changes, glucose utilization was decreased bilaterally in all parts of the cerebral cortex, and ipsilaterally in the thalamus. In the hippocampus, CA1 metabolic activity was depressed at day 0 and increased at day 15 while CA3 glucose utilization was increased at both day 0 and 15. By day 30, there were almost no pyramidal cells left in the two hippocampal regions. At day 120, ipsilateral decreases persisted in the entorhinal cortex, anterior and ventromedian thalamus, and metabolic increases were recorded bilaterally in the central amygdala, anterior hypothalamus and mamillary body. At all times after kainate, a normo-, hypo- or hypermetabolic level was recorded in the dentate gyrus. The present study shows that the process of hippocampal sclerosis involves bilateral cortical reactivity and the participation of some limbic forebrain and motor structures. When hippocampal sclerosis has fully developed, hypometabolism is limited to regions directly connected to the damaged hippocampus and most likely involved in the new hyperexcitable circuit of limbic seizures. PMID- 10678751 TI - Gender differences in brain volume and size of corpus callosum and amygdala of rhesus monkey measured from MRI images. AB - While it has been established that the weight of the female rhesus monkey brain is less than that of the male, the sexual dimorphism of specific brain structures has not been well-documented. To further understand potential sex differences, we measured the whole brain volume and the size of the corpus callosum (mid sagittal) and amygdala (largest coronal section) in MRI images from juvenile to adult male and female rhesus monkeys between 8 months and 7.2 years of age. The mean volume of the male brain was 89.2 +/- 1.9 (S.E.M.) compared to the female brain volume of 70.8 +/- 0.72 cm3. The average area of the corpus callosum increased from 8 months to 4.5 years; 0.56 to 0.93 cm2 in males and 0.45 to 0.66 cm2 in females. However, the average area of splenium is significantly greater in females (0.280 cm2), than males (0.184 cm2). The average area of the amygdala did not change with age; it was 1.07 +/- 0.037 (S.E.M.) in males and 1.08 +/- 0.022 cm2 in females. This data suggests that the whole brain volume and the size of the entire corpus callosum of young adult female rhesus monkeys are approximately 20% smaller than those of young adult males. Interestingly, the area of the splenial portion of the corpus callosum is larger in female monkeys. The size of the amygdala showed no sex difference. PMID- 10678752 TI - Evidence that synaptically-released zinc contributes to neuronal injury after traumatic brain injury. AB - Prior evidence indicates that synaptically-released zinc enters postsynaptic neurons in toxic excess during ischemia and seizures. In addition, prevention of this zinc translocation has been shown to be neuroprotective in both ischemia and seizures. Here we show evidence that the same translocation of zinc from presynaptic boutons into postsynaptic neurons occurs after mechanical injury to the brain. Specifically, using a rat model of traumatic brain injury, we show that trauma is associated with (i) loss of zinc from presynaptic boutons (ii) appearance of zinc in injured neurons, and (iii) neuroprotection by intraventricular administration of a zinc chelator just prior to brain impact. The possible use of zinc chelators for neuroprotection after head trauma is considered. PMID- 10678753 TI - Histochemically-reactive zinc in amyloid plaques, angiopathy, and degenerating neurons of Alzheimer's diseased brains. AB - Excess brain zinc has been implicated in Alzheimer's neuropathology. Here we evaluated that hypothesis by searching the brains of Alzheimer's patients for abnormal zinc deposits. Using histochemical methods, we found vivid Zn2+ staining in the amyloid deposits of dense-core (senile) plaques, in the amyloid angiopathy surrounding diseased blood vessels, and in the somata and dendrites of neurons showing the characteristic neurofibrillary tangles (NFT) of Alzheimer's. In contrast, brains from age-matched, non-demented subjects showed only occasional staining for Zn2+ in scattered neurons and possible plaques. A role of abnormal zinc metabolism in Alzheimer's neuropathology is suggested. PMID- 10678754 TI - p75-mediated neuroprotection by NGF against glutamate cytotoxicity in cortical cultures. AB - Accumulating evidence suggests that the neurotrophin receptors, Trks and p75, play distinct roles in regulating cells survival and death, with Trks important for cell survival, and p75 acting to induce cell death. Here, we provide evidence that, in neuronal cultures from rat cerebral cortex, nerve growth factor (NGF) exerts neuroprotective actions via p75. Incubating cultures with NGF for 1-24 h protected cortical neurons from delayed cytotoxicity induced by brief exposure to glutamate. Delayed neurotoxicity induced by a calcium ionophore, ionomycin, or nitric oxide (NO) donors such as S-nitrosocysteine (SNOC) and 3 morpholinosydnonimine (SIN-1), was also attenuated by pretreatment with NGF. RT PCR analysis revealed the presence of p75 and trkB transcripts in cortical cultures, but did not detect transcripts of trkA, a high-affinity receptor for NGF. Brain-derived neurotrophic factor (BDNF), but not NGF, induced tyrosine phosphorylation of Trks, indicating that NGF does not activate Trks in cortical neurons. Concurrent application of anti-p75 neutralizing antibody markedly reduced the neuroprotective effect of NGF, but resulted in only a modest reduction of that of BDNF. BDNF-induced neuroprotection, but not NGF-induced neuroprotection, was inhibited by a protein synthesis inhibitor cycloheximide. Distinct signaling pathways mobilized by NGF and BDNF were also revealed in that NGF but not BDNF stimulated significant production of ceramides, whereas BDNF but not NGF caused persistent activation of mitogen-activated protein kinases. These results indicate that, although NGF and BDNF both protect cortical neurons from excitotoxicity, the mechanisms involved in their effects are totally different. The present results are, to our knowledge, the first to demonstrate the principal involvement of p75 in cytoprotective actions of neurotrophins. PMID- 10678755 TI - Functional MRI of apomorphine activation of the basal ganglia in awake rhesus monkeys. AB - Functional magnetic resonance imaging (fMRI) was used to analyze blood oxygen level-dependent (BOLD) responses in the nigrostriatal system (caudate nucleus, putamen and substantia nigra) of awake rhesus monkeys to systemic apomorphine administration. The study (1) measured BOLD responses as an index of neuronal activity in the three structures following injections of the mixed D1/D2 agonist, and (2) assessed the effects of isoflurane anesthesia on the fMRI responses. Compared to control saline injections, 0.1 mg/kg apomorphine significantly activated the caudate nucleus (P < or = 0.005), putamen (P < or = 0.001) and substantia nigra (P < or = 0.005). The responses were consistent with activation of GABAergic neurons in these three structures seen in other animal models. Isoflurane gas measurably blunted the response to apomorphine, so that a significant apomorphine activation was only seen in the substantia nigra of anesthetized animals. Even there, the mean MR signal change was reduced from 9.8% in awake monkeys to 2.3% in anesthetized animals. The data support the hypothesis that fMRI can be used to study the effects of drugs that alter basal ganglia activity in awake rhesus monkeys. PMID- 10678756 TI - Acrylamide-regulated neurofilament expression in rat pheochromocytoma cells. AB - Using the rat pheochromocytoma cell line (PC12), we present molecular evidence that the neurotoxicant acrylamide directly induces neurofilament gene expression, and the signaling pathways are initially distinctive from, but eventually merged into, that for nerve growth factor (NGF)-induced neurofilament expression. In PC12 cells, acrylamide increased neurofilament protein levels and synthesis. Acrylamide had no effect on the stability of neurofilament mRNAs suggesting that it directly increased neurofilament mRNA synthesis. K252a, a selective inhibitor for NGF receptor gp140trk, had no effect on acrylamide induction, but completely inhibited NGF-induced neurofilament protein synthesis. Therefore, the initial step for acrylamide signaling was distinctive from NGF. Dexamethasone reversed the effects of both NGF and acrylamide on neurofilament protein levels and synthesis indicated that there is a dexamethasone-sensitive signaling step upon which NGF and acrylamide merge, suggesting involvement of transcription activating proteins like AP-1. These results, taken together with previous studies of transgenic mice that overexpress neurofilament genes, may partially explain the mechanisms of neurofilament accumulation in distal parts of large axons, a pathognomonic feature of acrylamide neurotoxicity in animals. PMID- 10678757 TI - Neuregulin found in cultured-sciatic nerve conditioned medium causes neuronal differentiation of PC12 cells. AB - The present work deals with the search and identification of the molecule or combination of molecules, present in a medium conditioned by cultured rat-sciatic nerves (CM), able to cause neuronal differentiation of PC12 cells. The molecular mass range of the active fraction, as well as the thermostability and heparin affinity of the active component found in previous work, all characteristics shared with neuregulin (NRG) family members, led us to search for a NRG protein in the CM. Nerves were previously cultured for 8 days and the CM collected every 24 h, the following 3 days. The CM was concentrated (30,000 NMWL) and fractionated by quaternary ammonium chromatography and Cibacron blue affinity chromatography. The most active fraction B1.2 was further characterized by heparin affinity chromatography, size exclusion HPLC, Western blotting and immunoprecipitation. Results reveal abundance of NRG mRNA in the cultured nerves, presence of a 54 kDa NRG protein in the CM that increases along fractionation, and progressive diminution of fraction B1.2 differentiation activity on PC12 cells by gradual removal of the NRG protein by immunoprecipitation. The abundance of Schwann cells and the lack of axons in the cultured nerves suggest Schwann cells as the main NRG source, to which fibroblasts and perineurial cells might contribute. PMID- 10678758 TI - Nitrotyrosine generation via inducible nitric oxide synthase in vascular wall in focal ischemia-reperfusion. AB - Nitrotyrosine produced by NO-mediated reaction is a possible marker for cytotoxicity in brain ischemia. In this study, we aimed to determine whether iNOS is responsible for the nitrotyrosine formation and which type of cell is predominantly nitrated. Fifty-eight wild-type and 28 iNOS knockout male mice were used. Under halothane anesthesia the left middle cerebral artery was occluded for 2 h and reperfused for 0.5 or 15 h. The ratio of nitrotyrosine to total tyrosine (%NO2-Tyr) was measured by means of a hydrolysis/HPLC. After 0.5-h reperfusion, %NO2-Tyr in the ischemic cortex of wild-type and knockout mice amounted to 0.037 +/- 0.040% (n = 8) and 0.064 +/- 0.035% (n = 6), respectively, being significantly higher than that in the sham operation group (n = 7) (P < 0.05). After 15-h reperfusion, nitrotyrosine was detected only in wild-type mice (0.039 +/- 0.025%, n = 7), not in knockout or sham-operated mice (P < 0.05). Immunohistochemical reaction for nitrotyrosine was seen predominantly in the vascular wall in the peri-infarct region of the cerebral cortex in wild-type mice after 15-h reperfusion, but not in corresponding knockout mice. Our data suggest that iNOS is responsible for nitrotyrosine formation in the later phase of reperfusion, and that vascular endothelium is the major site of this reaction, at least in the case of 15-h reperfusion. PMID- 10678759 TI - Oxidative stress and Ca2+ influx upregulate calpain and induce apoptosis in PC12 cells. AB - Calpain, a Ca2+-dependent cysteine protease, has previously been implicated in apoptosis or programmed cell death (PCD) in immune cells. Although oxidative stress and intracellular free Ca2+ are involved in neurodegenerative diseases, the mechanism of neuronal cell death in the central nervous system (CNS) due to these agents has not yet been defined. To explore a possible role for calpain in neuronal PCD under oxidative stress and Ca2+ influx, we examined the effects of H2O2 and A23187 on PC12 cells. Treatments caused PCD (light microscopy and TUNEL assay) with altered mRNA expression (RT-PCR) of bax (pro-apoptotic) and bcl-2 (anti-apoptotic) genes, resulting in a high bax/bcl-2 ratio. Control cells expressed 1.3-fold more microcalpain (requiring microM Ca2+) than mcalpain (requiring mM Ca2+). Expression of mcalpain was significantly increased following exposure to oxidative stress and Ca2+ influx. The mRNA levels of calpastatin (endogenous calpain inhibitor) and beta-actin (house-keeping) genes were not changed. Western analysis indicated degradation of 68 kDa neurofilament protein (NFP), a calpain substrate. Pretreatment of cells with MDL28170 (a cell permeable and selective inhibitor of calpain) prevented increase in bax/bcl-2 ratio, upregulation of calpain, degradation of 68 kDa NFP, and occurrence of PCD. These results suggest a role for calpain in PCD of PC12 cells due to oxidative stress and Ca2+ influx. PMID- 10678760 TI - Subcommissural organ-Reissner's fiber complex of the teleost Clarias batrachus responds to GABA treatment. AB - Subcommissural organ (SCO) is a highly specialized ependymal gland located in the roof of the third ventricle. The secretory products of the SCO, which condense to form Reissner's fiber (RF), were recently found to cross-react with the anti calcitonin antibody. To understand the mechanisms regulating the formation of the RF and the possible function of these discrete structures, we studied the response of the SCO-RF complex to intracranially administered GABA, using immunocytochemical labeling with anti-calcitonin antibody. Although the SCO-RF complex of control fish was intensely immunostained, 1 h after GABA treatment, the ependymal cells revealed partial loss of immunoreactivity; the RF showed occasional loss of immunoreactivity with its diameter increased by about 56% of the control value. Following 2 h of GABA treatment, the SCO revealed dramatic loss of calcitonin-like immunoreactivity from the ependymal cells. The RF showed a dual response in this group, while in some segments the RF appeared conspicuously thick, elsewhere it appeared thin. The mean diameter was, however, not significantly different from the normal. Following 4 h of GABA treatment, while calcitonin-like immunoreactive material made its reappearance in the SCO, the RF diameter was uniformly reduced to about 35% of the control value. The responses by the RF as well as the SCO to intracranially administered GABA were blocked by pretreatment with bicuculline, a GABA(A) receptor antagonist. The results suggest that GABA, acting via GABA(A) receptors, may trigger the release of secretory material from the SCO and induce histomorphological changes in the RF indicative of discharge of stored material. PMID- 10678761 TI - Propofol increases agonist efficacy at the GABA(A) receptor. AB - Using the whole-cell patch-clamp technique, we have determined that propofol, but not midazolam, increases the efficacy of piperidine-4-sulphonic acid (P4S), a partial agonist at alpha1beta1gamma2s, GABA(A) receptors expressed in HEK 293 cells. These findings are consistent with the idea that propofol facilitates receptor gating, while midazolam increases receptor occupancy by the agonist. PMID- 10678762 TI - Macrophages: a major source of cytochrome b558 in the rat carotid body. AB - The carotid body monitors arterial oxygen tension. Spectrophotometric recording of the intact organ has revealed a cytochrome aa3 and a cytochrome b558 as potential oxygen sensor candidates. The latter is known as part of the NADPH oxidase system generating superoxide anions in the "respiratory burst" defense mechanism, and glomus cells have been found to exhibit immunoreactivity against this phagocyte cytochrome b558. Using a monoclonal antibody against the large cytochrome b558 subunit, gp91phox, and other antibodies serving as neural (PGP 9.5) and monocyte/macrophage markers (ED1, ED2), we here demonstrate at light and electron microscopical level that monocytes/macrophages are abundantly present in the rat carotid body and represent the major source of cytochrome b558 in this organ. Their presence has profound implications on the interpretation of spectrophotometric recordings aimed to elucidate the mechanisms of oxygen sensing since their high cytochrome b558 content will obscure possible contributions of cell types involved in the oxygen sensor process. PMID- 10678763 TI - Neurotrophin receptors in the somatosensory cortex of the mature rat: co localization of p75, trk, isoforms and c-neu. AB - Trk immunoreactivity is expressed by a discrete population of cortical neurons, primarily those with cell bodies in layer Vb and dendrites in supragranular cortex. We tested the hypothesis that neurons co-express multiple isoforms of trk receptors. The distribution of neurons expressing specific high affinity neurotrophin receptors was determined immunohistochemically. Multiple antibodies directed against each trk isoform and an antibody directed against an epitope shared by all three trk isoforms were used. The distribution of neurons expressing each of the three receptors was virtually identical. Each anti-trk antibody primarily labeled neurons with cell bodies in layer V. More than one third of layer V neurons was positive for a high affinity trk receptor. Few immunoreactive somata (1%-5%) were in the other layers. In addition, the neuropil in the supragranular laminae was immunopositive for each trk isoform. Recent data show that layer V neurons in the mature somatosensory cortex express the tyrosine kinase receptor c-erbB2, also known as c-neu. Immunofluorescence double labeling shows that approximately 80% of the c-neu-immunolabeled neurons in layer V co expressed pan-trk immunoreactivity and two-thirds of all c-neu-positive neurons expressed a specific trk isoform. We concluded from these data that there is significant co-expression of trk isoforms in layer V neurons. In summary, trkA, trkB, trkC, and c-neu were primarily expressed by cortical projection neurons in layer V and co-expression among these receptors was common. This implies that cortical growth factor systems are redundant and that cortical neurons are responsive to more than one growth factor. PMID- 10678764 TI - In vivo evidence that activation of tyrosine kinase is a trigger for lipopolysaccharide-induced fever in rats. AB - We measured the rectal temperature of free-moving, conscious rats after intracerebroventricular (i.c.v.) injections of lipopolysaccharide (LPS) and interleukin-1beta (IL-1beta) with or without various antagonists to investigate the mechanisms involved in LPS-induced fever. LPS (3 microg) elicited significant increases in rectal temperature, which lasted from 0.5 h to more than 8 h after administration. This febrile response was inhibited by pretreatment with L-nitro arginine (LNA), indomethacin (IND), genistein (GEN), tyrphostin 46 and anti-rat IL-1beta antibody (anti-IL-1beta Ab), but was not inhibited by pretreatment with daidzein or chelerythrine (CHE) into the ventricle. LPS (0.3 microg) following orthovanadate (i.c.v.) produced fever, although the small amount of LPS (0.3 microg) or orthovanadate alone showed no effect on rectal temperature. I.c.v. injections of IL-1beta also induced fever of approximately 4-h duration. This effect was inhibited by pretreatment with IND and anti-IL-1beta Ab, but was not inhibited by pretreatment with LNA, GEN or CHE into the ventricle. These findings demonstrate that in the central nervous system, LPS increases IL-1beta production after activation of tyrosine kinase and NO synthase, and IL-1beta promotes prostaglandin production resulting in increased rectal temperature. Activation of tyrosine kinase in the central nervous system is probably a trigger for the febrile response induced by LPS. PMID- 10678765 TI - GSH transport in human cerebrovascular endothelial cells and human astrocytes: evidence for luminal localization of Na+-dependent GSH transport in HCEC. AB - The purpose of the present study was to identify and localize glutathione (GSH) transport in an in vitro tissue culture model of blood-brain barrier (BBB). The localization of Na+-dependent GSH transport in an immortalized cell line of human cerebrovascular endothelial cells (HCEC) and asymmetry of transport in Transwell studies were investigated. Initial studies with cultured HCEC established a significant (45%) Na+-dependency for GSH uptake in cultured HCEC pretreated with acivicin, an inhibitor of gamma-glutamyltranspeptidase (GGT). Transendothelial electrical resistance (TEER) and uptake of [35S]GSH from luminal and abluminal fluids of HCEC were measured in Na+-containing and Na+-free (choline chloride) buffers using cells grown on gelatin-coated membrane filters. TEER of HCEC monolayers in regular medium was 40.1 +/- 8.0 ohms cm2. Human astrocyte conditioned medium (ACM) caused no change in TEER, but increased GGT activity approximately threefold when measured in cell lysates. Luminal and abluminal GSH uptake increased in a time-dependent fashion and were not affected by inhibition of GGT activity with acivicin. Sodium dependency was only observed for luminal uptake (Na+-containing 2.41 +/- 0.15 vs. Na+-free 0.96 +/- 0.03 pmol/30 min/million cells, p < 0.001) but not for abluminal uptake (1.02 +/- 0.13 vs. 1.11 +/- 09, p > 0.05). Apparent efflux via the luminal membrane was lower in the presence of sodium as compared to that without sodium, further suggesting that a Na+-dependent uptake process for GSH is operative at this membrane. GSH uptake and efflux were also demonstrated in neonatal rat and fetal human astrocytes, both exhibiting partial Na+-dependency of uptake. In conclusion, our results show for the first time, that HCEC and astrocytes take up GSH by both Na+-dependent and -independent mechanisms. The Na+-dependent GSH transport process in HCEC appears to be localized to luminal plasma membranes of HCEC. PMID- 10678766 TI - The neural code for taste in the nucleus of the solitary tract of the rat: effects of adaptation. AB - Adaptation of the tongue to NaCl, HCl, quinine or sucrose was used as a tool to study the stability and organization of response profiles in the nucleus of the solitary tract (NTS). Taste responses in the NTS were recorded in anesthetized rats before and after adaptation of the tongue to NaCl, HCl, sucrose or quinine. Results showed that the magnitude of response to test stimuli following adaptation was a function of the context, i.e., adaptation condition, in which the stimuli were presented. Over half of all taste responses were either attenuated or enhanced following the adaptation procedure: NaCl adaptation produced the most widespread, non-stimulus-selective cross-adaptation and sucrose adaptation produced the least frequent cross-adaptation and the most frequent enhancement of taste responses. Adaptation to quinine cross-adapted to sucrose and adaptation to HCl cross-adapted to quinine in over half of the units tested. The adaptation procedure sometimes unmasked taste responses where none were present beforehand and sometimes altered taste responses to test stimuli even though the adapting stimulus did not itself produce a response. These effects demonstrated a form of context-dependency of taste responsiveness in the NTS and further suggest a broad potentiality in the sensitivity of NTS units across taste stimuli. Across unit patterns of response remained distinct from each other under all adaptation conditions. Discriminability of these patterns may provide a neurophysiological basis for residual psychophysical abilities following adaptation. PMID- 10678767 TI - Activation of CB1 cannabinoid receptors inhibits neurotransmitter release from identified synaptic sites in rat hippocampal cultures. AB - The effects of cannabinoids on synaptic transmission were measured optically in rat hippocampal cultures. Synaptic release sites were labeled with the fluorescent dye FM1-43 in a stimulus-dependent manner. Action potential-induced release of FM1-43 required extracellular Ca2+ and was inhibited 65 +/- 3% by blockade of high-threshold voltage-gated Ca2+ channels with omega-grammotoxin SIA (300 nM). The cannabimimetic drug, Win 55212-2 (300 nM), inhibited FM1-43 release by 51 +/- 3%. The inhibition produced by Win55212-2 was blocked by the CB1 cannabinoid receptor antagonist, SR141716 (1 microM). The intensity of FM1-43 labeled puncta ranged 4-fold, although the inhibition produced by Win55212-2 was distributed normally across synaptic sites of various labeling intensities. The FM1-43-based optical method appears promising for the study of the effects of cannabinoids and other drugs on synaptic networks. These results indicate that cannabimimetics act presynaptically to inhibit the release of neurotransmitter and that this inhibition is observed uniformly at boutons of varied activity levels. PMID- 10678768 TI - Evidence for physiological asymmetries in the intertectal connections of the pigeon (Columba livia) and their potential role in brain lateralisation. AB - In pigeons, visual object processing is lateralised with a dominance of the left tectofugal system. To test the hypothesis, that avian visual lateralisation may arise, at least in part, from asymmetric interhemispheric inhibition, the intertectal modulation was quantified in 19 pigeons. Field potentials were recorded from intratectal electrodes in response to a stroboscope flash to the contralateral eye. Electrical stimulation of the contralateral tectum changed these flash-evoked potentials. This change was taken as a measure of intertectal modulation. It was found that the left-to-right tectotectal modulation was more pronounced than vice versa, supporting the hypothesis of an asymmetric modulation between the tecta of both hemispheres. It is conceivable that this lateralised interhemispheric crosstalk could constitute an important component of asymmetric visual processing. PMID- 10678769 TI - Functional properties of the primary motor cortex and ventral premotor cortex in the monkey during a visually guided jaw-movement task with a delay period. AB - This study investigated single neuronal activity in the face area of the primary motor cortex (MI) and ventral part of the premotor cortex (PMv) while a monkey performed a visually guided jaw-movement task with a delay period. When the monkey executed the jaw movements, 48 MI and 53 PMv neurons showed statistically significant activities time-locked to jaw movements and were defined as movement related neurons. The activities of movement-related neurons could be classified into phasic, phasic-tonic and tonic patterns based on the changes in discharge rate. Most of the neurons exhibiting phasic and phasic-tonic activities probably contributed to the initiation of jaw movements, since they exhibited transient responses immediately after the onset of the go-cue indicating the jaw movement. In contrast, the sustained activity of the movement-related neurons exhibiting phasic-tonic and tonic activities may be involved in controlling and/or maintaining jaw position. Sustained activity was also detected during the delay period in 4 MI and 29 PMv neurons and these neurons were defined as set-related neurons. It is thought that these set-related neurons are involved in the preparation for the subsequent jaw movement, since the masticatory muscles showed no significant changes during the delay period. These findings suggest that the MI may be involved predominantly in the initiation and control of jaw movements, and that the PMv may be involved in motor preparation, and may play a role as a higher-order motor area related to the initiation and control of jaw movements. PMID- 10678770 TI - Expression of c-fos-like immunoreactivity in the feline brainstem in response to isometric muscle contraction and baroreceptor reflex changes in arterial pressure. AB - This study compared whether activation of muscle ergoreceptor afferents caused by isometric muscle contraction, activation of baroreceptor afferents induced by i.v. infusion of phenylephrine, or baroreceptor afferent inactivation, caused by carotid artery occlusion, elicit similar patterns of c-Fos induction in brainstem areas. Adult cats were anesthetized with alpha-chloralose, and in each case, the experimental intervention caused an increase in the arterial blood pressure. There were two sets of control experiments: in both, animals underwent the same surgical procedures but then either remained at rest for the entire study, or the tibial nerve was stimulated, as in the contraction group, following muscle paralysis with tubocurarine. Following the procedures, animals rested for 90 min to allow neuronal expression of c-Fos. Control cats showed very little c-Fos immunoreactivity (c-Fos-ir) in the brainstem. Muscle contraction induced c-Fos-ir expression mainly in the nucleus tractus solitarius, lateral reticular nucleus, lateral tegmental field, vestibular nucleus, subretrofacial nucleus, spinal trigeminal tract and in a lateral region of the periaqueductal grey (P 0.5-1.0). The majority of the c-Fos-ir was found in brainstem areas contralateral to the contracted muscle. In addition, muscle contraction induced c-Fos-ir in the dorsal horns of spinal segments L6-S1 on the ipsilateral side of the spinal cord. Phenylephrine infusion caused c-Fos-ir expression in the nucleus tractus solitarius, spinal trigeminal tract, solitary tract, and dorsal motor nucleus of the vagus. No c-Fos-ir was apparent in the periaqueductal grey. Carotid occlusions induced c-Fos-ir expression in the area postrema, nucleus tractus solitarius, solitary tract, and spinal trigeminal tract. Expression was bilateral. Areas that exhibited c-Fos-ir correspond to sites previously reported to release various neuropeptides in response to muscle contraction or carotid occlusions. These results indicate that the exercise pressor reflex and baroreflex activate similar, but not completely identical, sites in the brainstem. PMID- 10678771 TI - (+/-)-Alpha-methyl-4-carboxyphenylglycine, a metabotropic glutamate receptor blocker, impairs retention of an inhibitory avoidance task in rats when infused into the basolateral nucleus of the amygdala. AB - The amygdala is important for memory processes of emotionally motivated learning and the amygdala glutamatergic system may play a key role in this process. In this study we assessed the effect of the infusion of (+/-)-alpha-methyl-4 carboxyphenylglycine (MCPG), a metabotropic glutamate receptor (mGluR) antagonist, into the basolateral complex of the amygdala (BLA) on the learning and retention of an emotionally motivated task. Rats received either vehicle or three different doses of MCPG (0.2, or 1.0, or 5.0 microg/0.2 microl/side, respectively) bilaterally into the BLA, 5 min before they were trained in a continuous multiple-trial inhibitory avoidance (CMIA) task. Response latencies during the training were recorded. Retention was assessed 8 days later. MCPG in the doses given did not significantly affect the acquisition of the CMIA task. However, MCPG at a dose of 5.0 microg/0.2 microl/side impaired the long-term retention test performance. Additionally, a nociception test indicated that dose of MCPG infused into the BLA did not affect the footshock sensitivity. Our results indicate that MCPG, when infused into the BLA of rats prior to the training, impaired long-term memory of aversive training without affecting acquisition. PMID- 10678773 TI - Expression and development of the proenkephalin mRNA in the C cells of chicken ultimobranchial glands. AB - A large number of enkephalin-immunoreactive cells transiently appear in chick ultimobranchial glands during embryonic development. The expression and development of proenkephalin mRNA were examined in the ultimobranchial glands by in situ hybridization with digoxigenin (DIG)-labeled oligonucleotide probes, in comparison with those of calcitonin mRNA and enkephalin peptide. Proenkephalin mRNA, as well as calcitonin mRNA, appeared in some C cells at embryonal day 14 (E 14), and in many cells at E 16. Subsequently, there is a marked increase in the level of calcitonin mRNA around E 18-19; all C cells exhibited intense reaction for calcitonin mRNA. After hatching, intensity of calcitonin mRNA expression was more and more increased. Northern blot analysis with the calcitonin probe also indicated that calcitonin synthesis of the C cells progressively increased with developmental gradient, and reached to the adult level at 1 month after hatching. On the other hand, intensity of hybridization signal of proenkephalin mRNA was maintained moderately during development. In contrast to enkephalin immunoreactivity, which is markedly decreased after hatching, proenkephalin mRNA expression was consistently detected in many C cells of 1- and 2-month-old chickens. Reverse transcription-polymerase chain reaction (RT-PCR) analysis confirmed that proenkephalin mRNA was obtained in the ultimobranchial glands of not only embryos but also 1-day- and 1-month-old chickens. Furthermore, Northern blot analysis demonstrated that a single band for proenkephalin mRNA was obtained in the poly (A)+RNA isolated from the ultimobranchial gland of 1-day-old chicks. Thus, the present study evidences that proenkephalin mRNA is synthesized in almost all C cells of chicken ultimobranchial glands throughout life. Enkephalin may be essential for C cell function. PMID- 10678772 TI - Co-distribution patterns of chromogranin B-like immunoreactivity with chromogranin A and secretoneurin within the human brainstem. AB - As members of the chromogranin family, chromogranin A, chromogranin B, and secretogranin II are acidic proteins found in large, dense core vesicles. They are endoproteolytically processed to smaller peptides and released after neuronal stimulation. Using immunocytochemistry, this study closely examines chromogranin B-like immunoreactivity within the human brainstem and then takes a comparative view of co-distribution patterns by chromogranin B, chromogranin A, and secretogranin II. We used an antiserum raised against a synthetic peptide (PE-11) present in the chromogranin B molecule. Secretogranin II was localized with an antiserum against secretoneurin, a 33 amino acid peptide, found within the secretogranin II precursor. Like chromogranin A and secretoneurin, chromogranin B is expressed through all levels of the human brainstem. Chromogranin B was exclusively detected in neuronal structures. The medial part of the substantia nigra pars reticulata, the nucleus interpeduncularis, the area of the central gray, and the raphe complex displayed a high density of PE-11-like immunoreactivity. Furthermore, a prominent staining was found in the medial, dorsal and gelatinous subnuclei of the solitary tract and the dorsal motor nucleus of vagus. The substantia gelatinosa of the caudal trigeminal nucleus and spinal cord were also very strongly PE-11-immunopositive. In conclusion, chromogranin B and secretogranin II showed similar distributions while neuronal localization typically differed from chromogranin A aside from a few exceptions. These findings may provide a framework for future research in revealing a functional role of chromogranin peptides in the human brainstem. PMID- 10678774 TI - A comparative non-radioactive in situ hybridization and immunohistochemical study of the distribution of alpha7 and alpha8 subunits of the nicotinic acetylcholine receptors in visual areas of the chick brain. AB - The distribution of mRNA transcripts corresponding to the alpha7 and alpha8 subunits of the nicotinic acetylcholine receptors (nAChRs) was studied in selected structures of the chick visual system with non-radioactive in situ hybridization and immunohistochemical techniques. The results indicated that the alpha7 and alpha8 nAChR transcripts are widely distributed in the brain, exhibiting differential expression in some structures but also some degree of co localization. The pattern of localization of alpha7 and alpha8 nAChR transcripts was highly correlated with immunohistochemical data, with very few instances of possible mismatches between the distribution of mRNAs and their corresponding proteins. PMID- 10678775 TI - Upregulation of brain somatostatin and neuropeptide Y following lidocaine-induced kindling in the rat. AB - Male Sprague-Dawley rats received a daily injection of 60 mg/kg of lidocaine (> 30 days). Twenty percent of rats developed convulsions (kindled rats) and remaining rats did not show convulsions (non-kindled rats). The level of immunoreactive somatostatin (IR-SRIF) in kindled rats was significantly increased in amygdala than that in non-kindled rats and control rats. Immunoreactive neuropeptide Y (IR-NPY) contents in kindled rats were significantly increased in amygdala, hippocampus, cortex and striatum compared to non-kindled and control rats. The expression of SRIF mRNA in kindled rats produced a significant increase in amygdala, while NPY mRNA in kindled rats showed an elevated expression in both amygdala and hippocampus. These results coincide with the previous findings with the elevated expression of SRIF and NPY mRNA in electrically and pharmacologically kindled models, suggesting the important role of these peptides in the kindling phenomenon. PMID- 10678776 TI - The distribution of neuropeptide Y and brain-derived neurotrophic factor immunoreactivity in hippocampal formation of the monkey and rat. AB - The distribution of neuropeptide Y (NPY) and Brain-Derived Neurotrophic Factor (BDNF) in the hippocampal formation of monkey and rat brains was studied immunohistochemically. The NPY-neuronal system is more highly developed in the monkey compared to that in the rat. The distribution of NPY-positive products was coincident with that of abundant BDNF-positive deposits. These observations suggest that the role of BDNF and the interaction of BDNF-NPY may differ between species. PMID- 10678777 TI - Sleep-inducing effects of adenosine microinjections into the medial preoptic area are blocked by flumazenil. AB - Microinjection of a wide range of sedative agents, including triazolam, pentobarbital, ethanol and adenosine, into the medial preoptic area has been shown to increase sleep, suggesting that it is an important (though not necessarily the only) anatomic site mediating hypnotic effects of these compounds. The mechanism by which adenosine increases sleep at this site is not clear, but one possibility is that this is related to its effects on the GABA(A) benzodiazepine receptor complex. In order to assess this possibility, this paper describes the administration of adenosine, alone and in combination with the benzodiazepine receptor blocker flumazenil, into the MPA. It was found that 12.5 and 25 nM of adenosine significantly reduced sleep latency and increased total sleep time. The sleep-inducing effect was blocked by flumazenil. Flumazenil caused a modest increase in total sleep, and prevented the increase in total sleep induced by the higher dose of adenosine. These data suggest that at least one aspect of the hypnotic properties of adenosine is mediated by a direct or indirect action on the GABA(A)-benzodiazepine receptor complex. PMID- 10678778 TI - Human interferon-alpha induces immobility in the mouse forced swimming test: involvement of the opioid system. AB - In a previous study, we indicated that human interferon (IFN)-alpha (IFN-alpha, 6 x 10(4) IU/kg, i.v.), but not human IFN-beta or -gamma, prolonged the immobility time of the forced swimming test in mice. In this study, we investigated the mechanism of the effect of human IFN-alpha. None of the mouse IFNs tested (IFN alpha/beta, IFN-beta, and IFN-gamma, 3 x 10(5) U/kg, i.v.) changed the immobility time or the spontaneous locomotor activity in mice. Indomethacin (10 mg/kg, s.c.), a cyclooxygenase inhibitor, did not affect the increase in the immobility time induced by human IFN-alpha (6 x 10(4) IU/kg, i.v.). However, naloxone (1 mg/kg, s.c.), an opioid receptor antagonist, blocked the increasing caused by human IFN-alpha in the forced swimming test. These results suggest that the increase in the immobility time caused by human IFN-alpha in the forced swimming test might be mediated through opioid receptors, but not mouse IFN receptors. PMID- 10678779 TI - Interactions of galanin and morphine in the spinal antinociception in rats with mononeuropathy. AB - The increased hind-paw withdrawal latency (HWL) to thermal stimulation and hind paw withdrawal threshold (HWT) to mechanical stimulation induced by morphine were attenuated by intrathecal injection of 1 or 3 nmol, but not 0.3 nmol of the selective galanin antagonist galantide. The result indicated a possible interaction between galanin and opioids in the transmission of presumed nociceptive information in the spinal cord of rats with mononeuropathy. PMID- 10678780 TI - Potassium chloride depolarization enhances MPP+-induced hydroxyl radical generation in the rat striatum. AB - We determined that extracellular potassium ion concentration, [K+]o-induced depolarization, enhances 1-methyl-4-phenylpyridinium ion (MPP+)-induced hydroxyl radical (*OH) generation in the rat striatum. Rats were anesthetized, and sodium salicylate in Ringer's solution (0.5 nmol/microl/min) was infused through a microdialysis probe to detect the generation of *OH as reflected by the non enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) in the striatum. Induction of high concentration KCl (70 mM) drastically increased formation of *OH trapped as DHBA by the action of MPP+. When dopamine (DA) was administered to the high KCl-treated animals, a marked elevation of DHBA was observed, compared with MPP+-only-treated animals, that showed a positive linear correlation between DA and *OH formation trapped as DHBA (R2 = 0.979) in the dialysate. When corresponding experiments were performed with iron (II), the same results were obtained: a positive linear correlation between the release of iron (II) and DHBA (R2 = 0.988) in the dialysate. These results suggest that [K+]o-induced depolarization enhances the formation of *OH products of efflux/oxidation due to MPP+. PMID- 10678781 TI - A role of glutamate in drug-induced ototoxicity: in vivo microdialysis study combined with on-line enzyme fluorometric detection of glutamate in the guinea pig cochlea. AB - The time course of the changes in perilymphatic glutamate was determined during the application of kanamycin and ethacrynic acid, which are known to damage the hair cells in the inner ear. For the continuous recording of glutamate, the microdialysis technique combined with an enzyme-linked fluorometric assay was used. In guinea pigs receiving a loading dose of 800 mg/kg of kanamycin subcutaneously followed 3 h later by an i.v. injection of 40 mg/kg of ethacrynic acid, a marked glutamate release was clearly found about 2 h after the injection of ethacrynic acid. Injection of kanamycin or ethacrynic acid alone did not produce any change in the perilymphatic glutamate. The morphological changes induced by the administration of both drugs indicated that the collapsing hair cells might release glutamate into the perilymphatic space. The present findings provide additional evidence that glutamate acts as an aggravating factor in aminoglycoside-induced ototoxicity. PMID- 10678782 TI - Loss of bag-1 immunoreactivity in rat brain after transient middle cerebral artery occlusion. AB - Although bag-1 is a strong apoptosis repressor protein, its functions in normal or injured brains are not fully understood. In the present study, we investigated expression of bag-1 protein in rat brain after transient middle cerebral artery (MCA) occlusion, and compared the results with that of terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling (TUNEL). Immunohistochemical analysis revealed that neuronal, choroid plexus, and ependymal cells were positively stained in the sham control brain. After 90 min of transient MCA occlusion, immunoreactivity for bag-1 progressively decreased from 3 to 48 h in the nuclei of neurons. Western blot analysis revealed that immunoreactive bag-1 was markedly decreased in the nuclear fraction. In contrast, cytosolic and mitochondrial fractions showed no or only slight change after the ischemia. TUNEL positive cells appeared at 48 h after the reperfusion, which was preceded by loss of bag-1 immunoreactivity. The present results suggest that bag-1 plays some roles in normal neuronal function, and its loss may be involved in neuronal cell death after ischemia. PMID- 10678783 TI - New advances in preventing and treating serious surgical infection: introduction. PMID- 10678784 TI - Antimicrobial action and pharmacokinetics/pharmacodynamics: the use of AUIC to improve efficacy and avoid resistance. AB - In in-vitro and in animal models, antibiotics show good relationships between concentration and response, when response is quantified as the rate of bacterial eradication. The strength of these in-vitro relationships promises their utility for dosage regimen design and predictable cure of human infections. Resistance is also predictable from these parameters, fostering a rational means of using dosing adjustments to avoid or minimize the development of resistant organisms. Newly developed computerized methods for the quantitation of susceptibility allow testing of integrated kinetic-susceptibility models in patients. Our attention has focused recently on fluoroquinolones, since they are relatively non-toxic and provide the necessary range of dosage needed to elucidate correlations between concentration and response in the Intensive Care Unit patient. Studies conducted in patients with nosocomial gram-negative pneumonia reveal good correlations between bacterial eradication and integration of concentration with bacterial susceptibility. In patients, the best correlation parameters are time over MIC, and the ratio of 24-hour AUC to MIC (AUIC). Patients with serious infections like nosocomial pneumonia require bactericidal antimicrobial activity. Studies in our laboratory demonstrate that the minimum effective antimicrobial action is an area under the inhibitory titer (AUIC) of 125, where AUIC is calculated as the 24-hour serum AUC divided by the MIC of the pathogen. This target AUIC may be achieved with either a single antibiotic or it can be the sum of AUIC values of two or more antibiotics. There is considerable variability in the actual AUIC value for patients when antibiotics are given in their usually recommended dosages. Examples of this variance will be provided using aminoglycosides, fluoroquinolones, beta-lactams, macrolides and vancomycin. The achievement of minimally effective antibiotic action, consisting of an AUIC of at least 125, is associated with bacterial eradication in about 7 days for beta-lactams and quinolones. When AUIC is increased to 250, the quinolone ciprofloxacin (which displays in vivo concentration dependent bacterial killing) can eliminate the bacterial pathogen in 1-2 days. Beta lactams, even when dosed to an AUIC of 250, often require longer treatment duration to eliminate the bacterial pathogen, because the in vivo bacterial killing rate is slower with beta-lactams than with the quinolones. This remains true even at AUIC values of 250 for both compounds, which is theoretically identical dosing. Antibiotic activity indices allow clinicians to evaluate individualized patient regimens. Furthermore, antibiotic activity is a predictable clinical endpoint with predictable clinical outcome. This value is also highly predictive of the development of bacterial resistance. Antimicrobial regimens that do not achieve an AUIC of at least 125 cannot prevent the selective pressure that leads to overgrowth of resistant bacterial sub populations. Indeed, there is considerable anxiety that conventional respiratory tract infection management strategies, which prescribe antibacterial dosages that may attain AUIC values below 125, are contributing to the pandemic rise in bacterial resistance levels. PMID- 10678785 TI - Antimicrobial chemotherapy in the control of surgical infectious complications. AB - In spite of the progress in surgical technique and antibiotic prophylaxis, postoperative infection still accounts for both the commonest surgical complication and one of the most frequent nosocomial infections, also causing an increase in duration and costs of hospital stay. The choice of treatment for post surgical infections requires an understanding of the usual infecting flora, available antimicrobial agents, and susceptibility patterns. The most common organisms in simple wound infection are gram-positive cocci and mainly Staphylococcus aureus. Staphylococcus epidermidis and S. aureus (quite often methicillin-resistant strains) are the organisms which predominate in the infectious complications following clean surgical procedures with implantation of vascular grafts or prosthetic devices. Mixed aerobic and anaerobic flora are mainly responsible for cases of intra-abdominal and intra-pelvic postoperative infections: the most common aerobes are Enterobacteriaceae (Escherichia coli, Proteus spp., and others) and enterococci, and among anaerobes Bacteroides fragilis group prevails. Adequate drainage and surgical control of the source of infection, when needed, and adjunctive effective antimicrobial therapy are important factors in successful treatment of postoperative infections. Semisynthetic penicillinase-resistant penicillins and glycopeptides (vancomycin and teicoplanin) are the drugs of choice for the treatment of infections caused by penicillin-resistant methicillin-sensitive, and methicillin-resistant, respectively, S. aureus and S. epidermidis. For the treatment of intra-abdominal and intra-pelvic infections, animal and human studies support the recommendation that treatment should be directed against both gram-negative enteric and anaerobic bacteria. Combinations of aminoglycosides with clindamycin or metronidazole have been widely used with great success; however adverse reactions such as nephrotoxicity and ototoxicity have been a problem in some patients. In recent years monotherapy with either a carbapenem (imipenem/cilastatin or meropenem) or a penicillin/beta-lactamase inhibitor combination has been proposed. Among these combination antimicrobials, piperacillin combined with tazobactam is a very well designed formulation. Indeed, piperacillin is active against a broad range of gram-negative and gram-positive pathogens, and tazobactam is a potent beta-lactamase inhibitor which acts on a variety of clinically important plasmid and chromosomal beta-lactamases. This combination seems particularly attractive for the treatment of mixed polymicrobial anaerobic aerobic infections such as intra-abdominal and intra-pelvic postoperative infections. PMID- 10678786 TI - Chemotherapy for gram-positive nosocomial sepsis. AB - Over recent years gram-positive bacterial pathogens have become dominant in many forms of nosocomial infections. The principal pathogens in severe infections are Staphylococcus aureus and enterococci. The utility of the traditional antibiotics used for nosocomial sepsis, particularly beta-lactam agents, has been severely compromised by the spread of resistance and there was, often, no therapeutic alternative to the glycopeptide antibiotics, vancomycin and teicoplanin, for empirical (and often also the specific) therapy of infections caused by methicillin-resistant S. aureus (MRSA) and Enterococcus spp. This reliance on glycopeptides, however, is now also threatened by acquired resistance. Vancomycin resistant enterococci (VRE), particularly E. faecium, have become a therapeutic problem in many European cities and are now endemic in some hospital wards. The recent reports from several continents of MRSA with reduced glycopeptide susceptibility (GISA) is of grave concern. New agents are needed to meet these threats and several classes of compounds are under development. One class is the streptogramins and the combination of quinupristin/dalfopristin (Synercid) is nearing licensing. Clinical trials and a compassionate use programme have already shown it to have considerable promise for the treatment of the most problematic forms of gram-positive nosocomial sepsis, including MRSA and vancomycin-resistant E. faecium infections that had failed therapy with other antibiotics. PMID- 10678787 TI - New advances in the use of antimicrobial agents in surgery: intra-abdominal infections. AB - Advances in both technical methods and antimicrobial therapy have significantly reduced morbidity and mortality for secondary (enterogenous) or community acquired intra-abdominal infections. Presumptive antimicrobial therapy for most community-acquired intra-abdominal infection can be safely initiated with a single broad-spectrum antimicrobial effective against the expected Enterobacteriaceae and anaerobic flora. Beta-lactams and carbapenems are effective against gram-negative rods and anaerobes, achieve therapeutic levels rapidly, and have low toxicity in the absence of penicillin allergy. Second generation cephalosporins (e.g. cefoxitin and cefotetan) remain useful in surgical prophylaxis and treatment of mild community-acquired pneumonia, but limitations in their spectra and antimicrobial resistance restrict their utility in more serious infections. The fourth generation cephalosporins are also effective, but should be combined with other antimicrobials such as metronidazole for adequate anaerobic coverage. Preliminary data on new fluoroquinolones are scant, but promising results were obtained in one clinical trial. We predict the current trend toward the use of broad-spectrum single agent antimicrobials for therapy of intra-abdominal infection will continue. PMID- 10678788 TI - Management of complicated intra-abdominal infections. AB - Complicated intra-abdominal infections are defined by the U.S. Food and Drug Administration as those in which an operation would not remove all of the infected tissue. Therefore perforated appendicitis, although usually straightforward to treat, would be considered complicated, whereas gangrenous non perforated appendicitis would not. Antibiotics play an adjunctive role to the surgical procedure in the management of these infections. Studies of newer antibiotics generally exclude critically ill patients, so it is unclear whether dose or duration of therapy can be addressed by such studies. Typical characteristics of anti-infective studies of intra-abdominal infection are: enrollment of upwards of 50% appendicitis cases, mortality 5%, and a clinical cure rate of 85%. Several antibiotic combinations with metronidazole are acceptable (e.g. third- or fourth-generation cephalosporin, aminoglycoside, aztreonam, or second-generation quinolone), as are several agents as monotherapy (e.g. second-generation cephalosporin, beta-lactamase agent, or third-generation quinolone). In addition to questions of dose and duration, questions have been raised regarding the value of intraoperative cultures, and whether issues of the quality of the surgical procedure can be addressed. The issue of the adequacy of surgical "source control" may be paramount, as an improper, untimely, or incorrect operation would have an overwhelmingly negative effect on outcome compared to the efficacy of the antibiotic. PMID- 10678789 TI - High-dose intravenous fluoroquinolones in the treatment of severe infections. AB - A bacterial infection should be considered "serious" in case of underlying disease, nosocomial origin, antibiotic resistant pathogen, and/or poor delivery of antibiotics at the site of infection. Treatment of most serious infections requires parenteral administration of antimicrobial agents. Intravenous fluoroquinolones are a class of antimicrobial agents from which physicians must choose when treating nosocomial infections. Fluoroquinolones are bactericidal antimicrobial agents that act by inhibiting DNA gyrase. They are active in vitro against most gram-negative bacteria and methicillin-susceptible staphylococci. Activity against anaerobic bacteria and streptococci is poor. The rapid development of bacterial resistance in centers with high quinolone usage is of great concern. Resistance develops most commonly in Pseudomonas aeruginosa and staphylococci. Most clinical trials with ciprofloxacin, ofloxacin, pefloxacin, the fluroquinolones currently available in France for parenteral use, are almost 10 years old. There are few studies with higher dosage and most of them have been carried out with ciprofloxacin. The findings of these studies indicate that higher dosage regimens of i.v. ciprofloxacin are much more effective against severe nososcomial infections than is the dosage of 200 mg twice daily. The higher dosage regimens resulted in greater rates of clinical cure and improvement in both monomicrobial and polymicrobial infections. Although the overall frequency of side effects to fluoroquinolones is low, seizures and allergic reactions have been attributed to their use. PMID- 10678790 TI - Fourth generation cephalosporins in the antimicrobial chemotherapy of surgical infections. AB - Surgical infections include a variety of entities such as secondary peritonitis, intra-abdominal abscesses, obstetric and gynecological infections as well as bone joint and soft-tissue infections. By definition the term "surgical infection" implies that surgery itself plays the major role in therapy, while antimicrobial chemotherapy is only supplementary. Broad-spectrum empirical regimens employed include the combination of a 1st or 2nd generation cephalosporin plus clindamycin or metronidazole +/- aminoglycoside (depending on the severity of the condition). Cefepime and cefpirome are new 4th generation parenteral cephalosporins with a spectrum of activity which makes them suitable for the treatment of infections caused by a wide variety of bacteria. They are active against both gram-positive and gram-negative organisms, including Staphylococcus aureus and Pseudomonas aeruginosa with activity comparable to or greater than that of cefotaxime or ceftazidime respectively. Cefepime in particular is also very active against strains of Enterobacter and Pseudomonas spp resistant to these two agents. In comparison with 3rd generation cephalosporins, cefepime appears to be less likely to induce resistance, due to a lower rate of hydrolysis by beta-lactamases, a low affinity for these enzymes and more rapid permeation into the cell. Despite the fact that a 4th generation cephalosporin is well-suited for the treatment of polymicrobial infections, the following should be kept in mind: (I) MRSA strains and Bacteroides fragilis group are not included in their spectrum of activity. (II) Cefpirome is the only cephalosporin with in vitro activity against Enterococci. (III) Severe surgical infections of nosocomial origin, and particularly in settings where Enterobacter spp predominate, represent the major indication for empirical use of a 4th generation cephalosporin in combination with a nitroimidazole. PMID- 10678791 TI - Treatment of fungal infections in surgical patients using conventional antifungals. AB - In surgical and intensive care units an alarming increase in the number of invasive fungal infections has been observed. This is partly due to temporal transferral of patients from hemato-oncological units or transplant units and partly to the enhanced use of corticosteroids and other immunosuppressants. Candida species have now become a common isolate in ill patients. Amphotericin B with or without flucytosine constituted the standard therapy for candidosis but similar response rates with less toxicity may be obtained with lipid and with fluconazole. Data on an improved outcome of candidemia if a central venous catheter is removed promptly are conflicting. Amphotericin B remains the standard therapy for other invasive mycoses; it is seldom possible to administer adequate doses and therefore the options and limitations of the triazoles and liposomal preparations should be explored. PMID- 10678792 TI - Role of newer azoles in surgical patients. AB - Fungal infection has become an important cause of morbidity and mortality in critically ill surgical patients. Surgical patients at highest risk for invasive mycoses include those undergoing extensive abdominal surgery, those with underlying malignancy or other immunosuppressive conditions, and patients undergoing transplantation. Nosocomial candidemia remains a major complication for patients in surgical intensive units; however, the epidemiology of invasive fungal infection continues to change with molds and yeasts other than Candida albicans emerging as important causes of infection especially in immunosuppressed patients. This changing epidemiology has resulted in the need for an expanded armamentarium of antifungal therapies. One effective approach has been the utilization of higher doses of well-tolerated azoles, such as fluconazole, particularly against yeasts with dose-dependent susceptibility. Alternatively, the presumptive use of therapeutic doses of fluconazole may be indicated in intensive care unit patients with persistent leukocytosis and fever in whom a source of fever cannot be identified, particularly if the patient is extensively colonized at mucosal sites with yeast. New azoles with an expanded spectrum of activity are in development. These include agents include voriconazole, which has activity against resistant yeasts and molds and is in phase III clinical trials, posaconazole (Sch 56592) and ravuconazole (BMS-207147)--both of which are less advanced in clinical development, but which also offer an expanded spectrum of activity. Other new azoles with expanded activity are still in the early phases of development. In this review, strategies for optimizing use of the clinically available new azoles and the potential for new agents are discussed. PMID- 10678793 TI - Impact of severe infections on the outcome of major liver surgery: a pathophysiologic and clinical analysis. AB - Although major progress has been achieved, particularly in the field of patient selection and postoperative intensive care, morbidity and mortality rates after major liver surgery are still significant. In fact, the mortality rate in major series reaches 30% of patients undergoing complex liver procedures, mostly related to postoperative septic events. Among these, although extra-abdominal infectious localizations are also frequently reported, intra-abdominal sepsis and abscess formation are probably the most frequent infective clinical presentations. The literature reports that the magnitude of the resection and duration of surgery are associated with a significantly higher postoperative morbidity and mortality rate. Severe postoperative infectious events cause a high proportion of this morbidity and in the presence of a septic evolution of the clinical picture the mortality rises dramatically. Such a tight association between severe infections and mortality after major hepatic surgery gives account to the fundamental role played by the liver in the metabolic homeostasis of the patient and also to the central hepatic function in the immune response to microorganisms of gastroenteric origin. After major liver surgery these central hepatic functions may by significantly impaired, thus leading to higher susceptibility to infections, in particular in the elderly. On these bases the improvement in prophylaxis protocols, in the early diagnosis and in the treatment of these postoperative infectious events can help optimize clinical results after major hepatic surgery. PMID- 10678794 TI - Emerging antimicrobial resistance in the surgical compromised host. AB - Improvements in the treatment of compromised patients have resulted in their prolonged survival in a debilitated state. Patients have repeated courses of antibiotics and become colonised with multiresistant pathogens during a stay in the intensive care unit. Surgical wound infections can then be very difficult to treat. Methicillin-resistant Staphylococcus aureus is now common although wide variations in prevalence exist between countries and regions. Klebsiella spp with multiple resistance is a common cause of septicemia and can be associated with cephalosporin use. Acinetobacter spp and vancomycin-resistant enterococci can cause infections resistant to all readily available antibiotics. The prevalence of infection with each of these pathogens is increasing. Control measures should include hand washing, universal precautions for infection control, source isolation, restrictive antibiotic policy and antibiotic rotation. Although new agents currently in trials may be effective in the long term, the future for antibiotic treatment or prophylaxis of surgical infections is in doubt. PMID- 10678795 TI - Management of the critically ill patient with severe sepsis. AB - Sepsis is a common cause of morbidity and mortality among the critically ill patient population. However, no anti-sepsis therapy has yet been found to be effective and treatment is thus largely supportive. Adequate fluid resuscitation must be accompanied by effective ventilation, and adrenergic agents may be needed to restore perfusion pressure and improve myocardial function. Enteral nutritional support with specialized nutrients has beneficial effects on morbidity, and should be started early. Further research will allow better definition of the septic patient according to immune status and enable more effective targeting of future anti-sepsis treatments. PMID- 10678796 TI - Antibiotic use and microbial resistance in intensive care units: impact of computer-assisted decision support. AB - As part of our integrated hospital information system (the HELP system), we developed computer-assisted decision support programs for antimicrobial prescribing that are available at bedside terminals throughout our 520-bed community hospital. Recently, options have been added to allow direct physician order entry of anti-infective agents in the shock-trauma intensive care unit (STRICU). Physicians prescribed the computer-suggested regimens for 46% but followed the suggested dose and interval for 93% of the orders during a 1-year study period. In comparison to a 2-year pre-intervention period, improved drug selection and reductions in adverse drug events and costs were seen. Antimicrobial resistance patterns for nosocomial gram-negative isolates remained stable or improved in the STRICU over an 11-year period of computer-assisted antibiotic management. We conclude that strategies for optimizing antimicrobial prescribing have the potential to stabilize resistance and reduce costs by encouraging heterogeneous prescribing patterns, use of local antimicrobial susceptibility patterns to inform empiric drug selection, and reduced "tonnage" of antibiotic use. PMID- 10678797 TI - Sepsis and septic shock: pro-inflammatory or anti-inflammatory state? AB - A considerable body of evidence indicates that together with an important pro inflammatory reaction, a anti-inflammatory response contributes to the onset of sepsis and organ failure. At a local site of injury or infection and during the initial appearance of pro- and anti-inflammatory mediators in the circulation, the beneficial effects of these compounds counterbalance their harmful effects. Only when the balance between these two forces is lost may these substances become harmful. The sequelae of an unbalanced systemic inflammatory reaction include derangement of microcirculation, shock, transudation into organs and defects of coagulation. An unbalanced systemic compensatory anti-inflammatory response often results in anergy and immunosuppression. The pro-inflammatory and anti-inflammatory conditions may ultimately reinforce each other, creating a state of destructive immunologic dissonance. In the present report, recent literature on cytokines was reviewed together with the 89 clinical papers published between 1993 and 1997 on the role of cytokines during sepsis and other inflammatory reactions. Cytokines were analyzed in more than 5,000 patients. Sepsis and septic shock were the two groups most represented (2834 individuals and 550 respectively). Only 12% (11) of the studies showed a correlation with the presence of sepsis or septic complications. Overall 27 (39%) studies found a correlation between levels of cytokine and mortality. Fifty (62%) of the 80 studies that investigated this correlation found that the amount of cytokines did not predict death. The rest of the 30 (48%) investigations depicted an unhomogeneous picture: even though 27 studies evidenced higher levels of cytokines in non-survivors, 3 studies found the opposite. PMID- 10678798 TI - New rationale for glucocorticoid treatment in septic shock. AB - Two recent small randomized trials evaluating a 5- to 12-day course of low dose hydrocortisone in patients with septic shock have reported a significant clinical improvement and a reduction in mortality. Recent studies indicate that an overaggressive and unregulated systemic inflammatory response is a major determinant of outcome in sepsis. In septic shock, nonsurvivors as opposed to survivors have over time: (1) significantly higher NF-kB activity in peripheral mononuclear cells, (2) persistent elevation in circulating inflammatory cytokine levels, and (3) elevated ACTH and cortisol levels. Current research recognizes that cytokines can cause a concentration-dependent resistance to endogenous glucocorticoids (GC). It is postulated that an excess of cytokine-induced transcription factors, such as NF-kB, may form complexes with activated glucocorticoid receptors (GCR), preventing GCR interaction with DNA. When T cells are incubated with a combination of cytokines, GC resistance is induced in a cytokine concentration-dependent fashion and reversed by removal of cytokines. Prolonged treatment with physiological doses of exogenous GCs may be necessary to compensate adequately for the inability of target organs to respond to endogenous cortisol and for the inability of the host to produce appropriately elevated levels of GCs. This hypothesis is supported by the laboratory findings of a recent randomized study of patients with unresolving acute respiratory disease. PMID- 10678799 TI - Short-term prophylaxis in clean-contaminated surgery. AB - Postoperative infections are not consistently controlled by current practice measures. From a recent study of 12,384 patients, postoperative infection occurred in 22% of colorectal procedures and 25% of upper gastrointestinal procedures. Infections were associated with a higher death rate, longer hospitalization, and more intense post-discharge care. Control of infections for clean-contaminated procedures requires effective bowel cleansing when appropriate, meticulous surgical technique, and timely antimicrobial administration. Many patients undergoing clean-contaminated surgery do not receive properly timed antimicrobials. Although the comparative value of oral (neomycin and erythromycin) or parenteral antimicrobials for colon surgery remains an unresolved issue, the combination can be beneficial for many colorectal operations. Third generation cephalosporins are not consistently more effective than older agents such as cefoxitin and increase bacterial resistance. Improper antimicrobial timing is one of the most common problems with surgical prophylaxis and is fully under the control of the surgeon. To maximize benefits of antimicrobial prophylaxis, systems should be devised to assure timely administration. PMID- 10678800 TI - Is single-dose antibiotic prophylaxis sufficient for any surgical procedure? AB - The objective of perioperative prophylaxis is to prevent postoperative infections, which are the primary cause of morbidity and mortality in patients undergoing surgery today. One cannot predict with certainty when bacterial contamination at the operative site may occur during surgery. Furthermore, it has been suggested that the period of highest risk may actually be at the end, rather than at the beginning, of the operation. Therefore, the effect of antimicrobial prophylaxis ideally should cover the entire perioperative "period of risk". It should be remembered that the period of risk for postoperative infection may last substantially longer than the actual surgical procedure. The duration of the risk period also may vary based on a number of other factors, such as the age and general condition of the patient, presence of concomitant disease, amount of blood loss during surgery, and number of blood transfusions required. Antimicrobial prophylaxis that provides coverage throughout the entire perioperative period of risk will reduce not only the risk of wound infections but may also reduce the danger of other types of infectious complications. Numerous clinical studies have clearly shown that appropriately-timed "single shot" prophylaxis is as effective as multiple-dose prophylaxis. This paper considers the evolution of this therapeutic intervention and reviews the opportunities available for antibiotic prophylaxis in surgery, with particular attention to the long-acting cephalosporin, ceftriaxone. PMID- 10678801 TI - Antimicrobial prophylaxis in surgery: the role of pharmacokinetics. AB - Even though surgical infection rates have decreased dramatically during the past 25 years, morbidity and mortality of infection in surgical treatment remains substantial. From a pharmacological point of view, the key factor of the efficacy of antibiotic prophylaxis is to attain bactericidal levels of antibiotic in serum and tissues (target site) during the whole intraoperative and early postoperative period. The success of antibiotic prophylaxis is assured only when the chosen antibiotic with a targeted spectrum and high antimicrobial efficacy is available at the critical moment, at the correct site and in sufficiently high concentration to prevent bacterial contamination of the surgical area. It would be desirable for reasons of convenience and cost if a single preoperative administration were sufficient. The pharmacokinetics and the half-life of antibiotics in the serum are directly related to the duration of activity of antibiotic in the tissue. Antibiotics with longer half-lives maintain levels in the tissues for longer periods than do antibiotics with shorter half-lives and they cover with a single dose the time required for prophylaxis even for longer operations. Finally, the application of the pharmacokinetic properties of antibiotics to surgical prophylaxis can provide the surgeon with certainty that adequate coverage and protection with antibiotics are achieved before and throughout the operation. PMID- 10678802 TI - Monitoring of antimicrobial prophylaxis in general surgery. AB - The incidence of infections in general surgery is related to different factors. Cost-benefit analysis of antimicrobic prophylaxis is positive, even though incorrect use may be even dangerous (development of resistance and/or superinfections, for instance). The authors report data on a study concerning a total of 316 patients divided into two series, who had antimicrobic prophylaxis before a surgical operation. 274 patients out of 316 (or 86.7%) had an ultra short (one-shot-only) or short (<24 hours) prophylaxis, 42 (13.3%) standard (>24 hours). The operations performed were classified following class of contamination, i.e. I (clean), II (potentially contaminated), III (contaminated). Antibiotics used were ceftizoxime, cefepime, ceftriaxone, piperacillin and gentamicin in combination. A total of 16 postoperative infections was observed (5%); 11 of these 16 belonged to class III operations. Escherichia coli and Staphylococcus aureus were isolated in most of the infected wounds. The data confirm what is reported in the literature. The authors conclude that a preoperative single-shot 3rd or 4th generation cephalosporin reduces the incidence of wound infections in clean and clean-contaminated surgery. PMID- 10678803 TI - Antimicrobial prophylaxis in obstetric and gynecological surgery. AB - A major problem in obstetric and gynecological surgery, especially following cesarean section in labor, total vaginal or abdominal hysterectomy, or myomectomy, is postoperative wound infection. Consequently, the use of antimicrobial prophylaxis for cesarean section and for gynecological surgery has been advocated and shown to be effective in reducing postoperative morbidity, costs and duration of hospitalization. We reviewed 1021 patients who underwent cesarean section (597 elective, 424 emergency) and 814 gynecological patients undergoing abdominal (373) or vaginal (248) hysterectomy and myomectomy (193) between 1997-98 in the Obstetrics and Gynecology Clinic of the University of Florence. Before surgery 83.6% of obstetric and 75.1% of gynecological patients received 1 or 2 g of a first or second generation cephalosporin i.v. as a single dose regimen at induction of anesthesia and sometimes a second postoperative dose. 1.5% of obstetric surgical patients had wound infection, as did 2.8% of gynecological surgery patients, with a mean postoperative hospital stay of 8 days. The short-term perioperative antimicrobial prophylaxis with cephalosporins is useful and provides the benefit of minimal toxicity and risk of chemoresistance. PMID- 10678804 TI - Management of Helicobacter pylori infection: gastroenterological and surgical perspectives. AB - Diagnosis and treatment of Helicobacter pylori is a crucial point in the management of the different gastroduodenal disorders. Management involves the general practitioner and different specialists such as internists, gastroenterologists and surgeons. Among the most frequent H. pylori-related gastroduodenal disorders of medical interest are some diseases such as dyspepsia and gastroesophageal reflux, where the role of the bacterium is not well defined and therefore the importance H. pylori eradication is still controversial. On the contrary, the relationship of H. pylori and gastric and duodenal peptic ulcer is widely and definitively proven, and there are no doubts regarding the importance of curing the bacterium in these disorders. However, the surgical aspect of peptic ulcer, in particular the relevance and management of its complications, has not been widely investigated so far. In fact, the prevalence of H. pylori in perforated, bleeding and stenotic peptic ulcers seem to be lower that in non complicated peptic ulcer, and whenever H. pylori eradication virtually prevents the re-bleeding of peptic ulcer in all cases, the effect of curing the bacterium in perforated and stenotic ulcers is still largely unknown. The management of H. pylori infection after gastric surgery is also still controversial. Most studies suggest that H. pylori can persist after gastric surgery whenever its incidence is much lower than that before operation. However it seems most unlikely that the infection plays a major role in the development of ulcer recurrence after gastric surgery or in the induction of gastric carcinoma. In any case, there are no convincing data that its cure may prevent the occurrence of gastric carcinoma following gastrectomy procedures. PMID- 10678805 TI - Helicobacter pylori determinants of pathogenicity. AB - The bacterium Helicobacter pylori colonises the stomach of man and induces a strong inflammatory response. Differences in the possession of pathogenicity determinants by H. pylori isolates could account in part for the different clinical outcomes of infection. The main H. pylori pathogenic factors, i.e. urease, the cytotoxin VacA, and the genes involved in virulence contained in the pathogenicity island (PAI) cag, may promote tissue damage and ulceration, and could contribute to gastric cancer development. Strains with the mosaic vacA allelic type s1a/m1 and possessing the cag insertion are considered endowed with increased inflammatory potential, and are more likely to be isolated from patients with peptic ulcer and gastric cancer. The presence in H. pylori cag PAI of operons involved in the stimulation of gastric epithelial cells to secrete high levels of inflammatory cytokines, in mobilisation of DNA, and formation of secretory mechanisms and conjugation apparati, could contribute to increase the risk of gastric cancer development in patients infected by this microorganism. PMID- 10678807 TI - Pet Travel Scheme: updated advice from MAFF. PMID- 10678806 TI - Helicobacter pylori: optimum diagnosis and test of cure. AB - The fact that about 50% of the world's population is infected with Helicobacter (H.) pylori and the important role that this bacterium plays in public health have been important incentives in the search for accurate diagnostic methods. A large number of invasive and non-invasive methods have been used to diagnose H. pylori infection. Each method has its advantages and disadvantages and each practitioner should choose the best diagnostic method according to the facilities available. Non-invasive tests for the diagnosis of H. pylori infection are largely used in clinical practice and in management of patients with gastroduodenal disease. Serology is the most widespread test but its use is not advised in the post-treatment follow-up. The Urea Breath Test is a simple, safe and highly accurate method ideal for evaluating the short-term follow-up of H. pylori eradication after therapy. PMID- 10678808 TI - Abomasal bloat, haemorrhage and ulcers in young Norwegian lambs. AB - An abomasal syndrome affecting mainly three- to four-week-old lambs was studied by using a case-control design involving 88 cases and 85 controls. The principal clinical signs were tympany and colic. The cases were divided into three groups according to the main gross pathological findings in 82 of them. The 28 lambs in group 1 had abomasal tympany, the 29 lambs in group 2 had severe damage to the abomasal mucosa, and the 25 lambs in group 3 consisted mainly of lambs with various other diseases in combination with abomasal changes. The lambs in group 1 had a significantly (P<0.05) lower mean (se) abomasal pH (2.7 [0.19]) than those in group 2 (4.1 [0.32]), group 3 (3.7 [0.39]) or the controls (3.3 [0.13]). The ruminal pH values ranged from 3.5 to 7.4 but there were no significant differences between the groups. Lambs with ulcers had a significantly (P<0.05) higher frequency of trichophytobezoars, than the cases without ulcers or the controls. The tympanic lambs in group 1 had a significantly higher mean packed cell volume, and higher mean red and white blood cell counts, and a significantly lower mean cell volume and mean cell haemoglobin concentration than the healthy control lambs. PMID- 10678809 TI - Identification of porcine circovirus in tissues of pigs with porcine dermatitis and nephropathy syndrome. AB - Thirty-three pigs affected by porcine dermatitis and nephropathy syndrome, 30 from Spain and three from the USA, were investigated in order to detect porcine circovirus (PCV) in their tissues. A standard in situ hybridisation technique using a specific DNA 317-bp probe based on a well-conserved sequence of PCV (which recognises both PCV-1 and PCV-2) was applied to formalin-fixed, paraffin embedded tissues. Twenty-eight of the 30 Spanish pigs and all three American pigs had PCV in at least one tissue. Viral nucleic acid was detected mainly in lymphoid organs, and especially the lymph nodes. The viral genome was also found, in order of decreasing quantity, in Peyer's patches, tonsil, lung, spleen, kidney, liver, and skin. Viral nucleic acid was located mainly within the cytoplasm of monocyte/macrophage lineage cells, including follicular dendritic cells, macrophages, histiocytes and Kupffer cells. No viral nucleic acid was found in damaged glomeruli or arteriolar walls. In frozen samples available from three Spanish pigs, the virus was identified as type 2 by using the polymerase chain reaction and restriction fragment length polymorphism. Most of the pigs from which serum was available were seropositive against porcine respiratory and reproductive syndrome virus (PRRSV), and PRRSV antigen was detected in the lung of two of the Spanish pigs. These results suggested that PCV is present in tissues of almost all pigs affected by PDNS, and PCV has to be considered as a possible agent involved in the pathogenesis of the syndrome. PMID- 10678810 TI - Urethral duplication and chromosomal translocation in a Swiss braunvieh heifer. AB - As it was urinating, a six-month-old Swiss braunvieh heifer produced a second stream of urine from a fistula that opened on the ventrolateral margin of the left vulval lip. A catheter was introduced into this opening and passed easily into the bladder. Urethrography showed that the fistula joined the urethra in the mid-pelvic region and that a single canal originated from the bladder. Endoscopy confirmed this finding and also revealed a duplication of the vaginal portion of the cervix, a division of the cranial vagina by a septum and a fibrous band in the region of the hymenal ring. Cytogenetic examination revealed reciprocal translocation between chromosomes 20q23 and 22q23. A diagnosis of urethra duplex, duplication of the vaginal portion of the cervix and reciprocal autosomal translocation between chromosomes 20 and 22 was made on the basis of these findings. PMID- 10678811 TI - Bordetella bronchiseptica infection in cats following contact with infected dogs. PMID- 10678812 TI - Dichloromethane poisoning in a dog: a case report. PMID- 10678813 TI - Birth of a BVDV-free calf from a persistently infected embryo transfer donor. PMID- 10678814 TI - Rabies in African wild dogs (Lycaon pictus) in the Madikwe Game Reserve, South Africa. PMID- 10678815 TI - Development of canine oocytes matured and fertilised in vitro. PMID- 10678816 TI - Chronic nasal discharge in dairy cows. PMID- 10678817 TI - Vet Helpline. PMID- 10678818 TI - Severe foot lesions in sheep. PMID- 10678819 TI - Society for Veterinary Clinical Research. PMID- 10678820 TI - Returning to practice. PMID- 10678821 TI - Reindeer concerns. PMID- 10678822 TI - Reindeer concerns. PMID- 10678823 TI - Interpreting differential temperature trends at the surface and in the lower troposphere AB - Estimated global-scale temperature trends at Earth's surface (as recorded by thermometers) and in the lower troposphere (as monitored by satellites) diverge by up to 0.14 degrees C per decade over the period 1979 to 1998. Accounting for differences in the spatial coverage of satellite and surface measurements reduces this differential, but still leaves a statistically significant residual of roughly 0.1 degrees C per decade. Natural internal climate variability alone, as simulated in three state-of-the-art coupled atmosphere-ocean models, cannot completely explain this residual trend difference. A model forced by a combination of anthropogenic factors and volcanic aerosols yields surface troposphere temperature trend differences closest to those observed. PMID- 10678824 TI - Crystal structure of the ribonucleoprotein core of the signal recognition particle. AB - The signal recognition particle (SRP), a protein-RNA complex conserved in all three kingdoms of life, recognizes and transports specific proteins to cellular membranes for insertion or secretion. We describe here the 1.8 angstrom crystal structure of the universal core of the SRP, revealing protein recognition of a distorted RNA minor groove. Nucleotide analog interference mapping demonstrates the biological importance of observed interactions, and genetic results show that this core is functional in vivo. The structure explains why the conserved residues in the protein and RNA are required for SRP assembly and defines a signal sequence recognition surface composed of both protein and RNA. PMID- 10678825 TI - Three-layered atmospheric structure in accretion disks around stellar-mass black holes AB - Modeling of the x-ray spectra of the Galactic superluminal jet sources GRS 1915+105 and GRO J1655-40 reveals a three-layered atmospheric structure in the inner region of their accretion disks. Above the cold and optically thick disk with a temperature of 0.2 to 0.5 kiloelectron volts, there is a warm layer with a temperature of 1.0 to 1.5 kiloelectron volts and an optical depth around 10. Sometimes there is also a much hotter, optically thin corona above the warm layer, with a temperature of 100 kiloelectron volts or higher and an optical depth around unity. The structural similarity between the accretion disks and the solar atmosphere suggests that similar physical processes may be operating in these different systems. PMID- 10678826 TI - Multidecadal changes in the vertical temperature structure of the tropical troposphere AB - Trends in global lower tropospheric temperature derived from satellite observations since 1979 show less warming than trends based on surface meteorological observations. Independent radiosonde observations of surface and tropospheric temperatures confirm that, since 1979, there has been greater warming at the surface than aloft in the tropics. Associated lapse-rate changes show a decrease in the static stability of the atmosphere, which exceeds unforced static stability variations in climate simulations with state-of-the-art coupled ocean-atmosphere models. The differential temperature trends and lapse-rate changes seen during the satellite era are not sustained back to 1960. PMID- 10678827 TI - Self-assembling amphiphilic siderophores from marine bacteria. AB - Most aerobic bacteria secrete siderophores to facilitate iron acquisition. Two families of siderophores were isolated from strains belonging to two different genera of marine bacteria. The aquachelins, from Halomonas aquamarina strain DS40M3, and the marinobactins, from Marinobacter sp. strains DS40M6 and DS40M8, each contain a unique peptidic head group that coordinates iron(III) and an appendage of one of a series of fatty acid moieties. These siderophores have low critical micelle concentrations (CMCs). In the absence of iron, the marinobactins are present as micelles at concentrations exceeding their CMC; upon addition of iron(III), the micelles undergo a spontaneous phase change to form vesicles. These observations suggest that unique iron acquisition mechanisms may have evolved in marine bacteria. PMID- 10678828 TI - Widespread complex flow in the interior of the antarctic ice sheet AB - It has been suggested that as much as 90% of the discharge from the Antarctic Ice Sheet is drained through a small number of fast-moving ice streams and outlet glaciers fed by relatively stable and inactive catchment areas. Here, evidence obtained from balance velocity estimates suggests that each major drainage basin is fed by complex systems of tributaries that penetrate up to 1000 kilometers from the grounding line into the interior of the ice sheet. This finding has important consequences for the modeled or estimated dynamic response time of past and present ice sheets to climate forcing. PMID- 10678829 TI - Density of phonon states in iron at high pressure AB - The lattice dynamics of the hexagonal close-packed (hcp) phase of iron was studied with nuclear inelastic absorption of synchrotron radiation at pressures from 20 to 42 gigapascals. A variety of thermodynamic parameters were derived from the measured density of phonon states for hcp iron, such as Debye temperatures, Gruneisen parameter, mean sound velocities, and the lattice contribution to entropy and specific heat. The results are of geophysical interest, because hcp iron is considered to be a major component of Earth's inner core. PMID- 10678830 TI - Inhibition of experimental liver cirrhosis in mice by telomerase gene delivery. AB - Accelerated telomere loss has been proposed to be a factor leading to end-stage organ failure in chronic diseases of high cellular turnover such as liver cirrhosis. To test this hypothesis directly, telomerase-deficient mice, null for the essential telomerase RNA (mTR) gene, were subjected to genetic, surgical, and chemical ablation of the liver. Telomere dysfunction was associated with defects in liver regeneration and accelerated the development of liver cirrhosis in response to chronic liver injury. Adenoviral delivery of mTR into the livers of mTR(-/-) mice with short dysfunctional telomeres restored telomerase activity and telomere function, alleviated cirrhotic pathology, and improved liver function. These studies indicate that telomere dysfunction contributes to chronic diseases of continual cellular loss-replacement and encourage the evaluation of "telomerase therapy" for such diseases. PMID- 10678831 TI - Prevention of acute liver failure in rats with reversibly immortalized human hepatocytes. AB - Because of a critical shortage in suitable organs, many patients with terminal liver disease die each year before liver transplantation can be performed. Transplantation of isolated hepatocytes has been proposed for the temporary metabolic support of patients awaiting liver transplantation or spontaneous reversion of their liver disease. A major limitation of this form of therapy is the present inability to isolate an adequate number of transplantable hepatocytes. A highly differentiated cell line, NKNT-3, was generated by retroviral transfer in normal primary adult human hepatocytes of an immortalizing gene that can be subsequently and completely excised by Cre/Lox site-specific recombination. When transplanted into the spleen of rats under transient immunosuppression, reversibly immortalized NKNT-3 cells provided life-saving metabolic support during acute liver failure induced by 90% hepatectomy. PMID- 10678832 TI - The glucocorticoid receptor: rapid exchange with regulatory sites in living cells. AB - Steroid receptors bind to site-specific response elements in chromatin and modulate gene expression in a hormone-dependent fashion. With the use of a tandem array of mouse mammary tumor virus reporter elements and a form of glucocorticoid receptor labeled with green fluorescent protein, targeting of the receptor to response elements in live mouse cells was observed. Photobleaching experiments provide direct evidence that the hormone-occupied receptor undergoes rapid exchange between chromatin and the nucleoplasmic compartment. Thus, the interaction of regulatory proteins with target sites in chromatin is a more dynamic process than previously believed. PMID- 10678833 TI - Dopaminergic loss and inclusion body formation in alpha-synuclein mice: implications for neurodegenerative disorders. AB - To elucidate the role of the synaptic protein alpha-synuclein in neurodegenerative disorders, transgenic mice expressing wild-type human alpha synuclein were generated. Neuronal expression of human alpha-synuclein resulted in progressive accumulation of alpha-synuclein-and ubiquitin-immunoreactive inclusions in neurons in the neocortex, hippocampus, and substantia nigra. Ultrastructural analysis revealed both electron-dense intranuclear deposits and cytoplasmic inclusions. These alterations were associated with loss of dopaminergic terminals in the basal ganglia and with motor impairments. These results suggest that accumulation of wild-type alpha-synuclein may play a causal role in Parkinson's disease and related conditions. PMID- 10678834 TI - Neuroimaging evidence for dissociable forms of repetition priming. AB - Repetition priming has been characterized neurophysiologically as a decreased response following stimulus repetition. The present study used event-related functional magnetic resonance imaging to investigate whether this repetition related response is sensitive to stimulus familiarity. A right fusiform region exhibited an attenuated response to the repetition of familiar stimuli, both faces and symbols, but exhibited an enhanced response to the repetition of unfamiliar stimuli. Moreover, both repetition effects were modulated by lag between successive presentations. Further experiments replicated the interactions between repetition, familiarity, and lag and demonstrated the persistence of these effects over multiple repetitions. Priming-related responses are therefore not unitary but depend on the presence or absence of preexisting stimulus representations. PMID- 10678835 TI - Sparse coding and decorrelation in primary visual cortex during natural vision. AB - Theoretical studies suggest that primary visual cortex (area V1) uses a sparse code to efficiently represent natural scenes. This issue was investigated by recording from V1 neurons in awake behaving macaques during both free viewing of natural scenes and conditions simulating natural vision. Stimulation of the nonclassical receptive field increases the selectivity and sparseness of individual V1 neurons, increases the sparseness of the population response distribution, and strongly decorrelates the responses of neuron pairs. These effects are due to both excitatory and suppressive modulation of the classical receptive field by the nonclassical receptive field and do not depend critically on the spatiotemporal structure of the stimuli. During natural vision, the classical and nonclassical receptive fields function together to form a sparse representation of the visual world. This sparse code may be computationally efficient for both early vision and higher visual processing. PMID- 10678836 TI - Mitochondrial FtsZ in a chromophyte alga. AB - A homolog of the bacterial cell division gene ftsZ was isolated from the alga Mallomonas splendens. The nuclear-encoded protein (MsFtsZ-mt) was closely related to FtsZs of the alpha-proteobacteria, possessed a mitochondrial targeting signal, and localized in a pattern consistent with a role in mitochondrial division. Although FtsZs are known to act in the division of chloroplasts, MsFtsZ-mt appears to be a mitochondrial FtsZ and may represent a mitochondrial division protein. PMID- 10678837 TI - Convergent solutions to binding at a protein-protein interface. AB - The hinge region on the Fc fragment of human immunoglobulin G interacts with at least four different natural protein scaffolds that bind at a common site between the C(H2) and C(H3) domains. This "consensus" site was also dominant for binding of random peptides selected in vitro for high affinity (dissociation constant, about 25 nanomolar) by bacteriophage display. Thus, this site appears to be preferred owing to its intrinsic physiochemical properties, and not for biological function alone. A 2.7 angstrom crystal structure of a selected 13 amino acid peptide in complex with Fc demonstrated that the peptide adopts a compact structure radically different from that of the other Fc binding proteins. Nevertheless, the specific Fc binding interactions of the peptide strongly mimic those of the other proteins. Juxtaposition of the available Fc-complex crystal structures showed that the convergent binding surface is highly accessible, adaptive, and hydrophobic and contains relatively few sites for polar interactions. These are all properties that may promote cross-reactive binding, which is common to protein-protein interactions and especially hormone-receptor complexes. PMID- 10678838 TI - Evidence for a high frequency of simultaneous double-nucleotide substitutions. AB - Point mutations are generally assumed to involve changes of single nucleotides. Nevertheless, the nature and known mechanisms of mutation do not exclude the possibility that several adjacent nucleotides may change simultaneously in a single mutational event. Two independent approaches are used here to estimate the frequency of simultaneous double-nucleotide substitutions. The first examines switches between TCN and AGY (where N is any nucleotide and Y is a pyrimidine) codons encoding absolutely conserved serine residues in a number of proteins from diverse organisms. The second reveals double-nucleotide substitutions in primate noncoding sequences. These two complementary approaches provide similar high estimates for the rate of doublet substitutions, on the order of 0.1 per site per billion years. PMID- 10678839 TI - Treating young patients with breast cancer. The evidence suggests that all should be treated with adjuvant therapy. PMID- 10678840 TI - Is CS gas dangerous? Current evidence suggests not but unanswered questions remain. PMID- 10678841 TI - Cholesterol and strokes. Cholesterol lowering is indicated for strokes due to carotid atheroma. PMID- 10678842 TI - Innovation to prevent dependency in old age. Technological innovations may reduce the cost burden of an ageing population. PMID- 10678843 TI - Beds in the NHS. The National Bed Inquiry exposes contradictions in government policy. PMID- 10678844 TI - Government inquiry finds inadequate beds provision. PMID- 10678845 TI - Police question medical MP over "euthanasia". PMID- 10678846 TI - Clinic sued for unauthorized use of sperm. PMID- 10678847 TI - In brief PMID- 10678848 TI - Senators introduce bill to improve patient safety. PMID- 10678849 TI - Doctor's book shames French prisons. PMID- 10678850 TI - GMC speeds up disciplinary action against doctors. PMID- 10678851 TI - GMC reprimands doctor for denigrating rival's service. PMID- 10678852 TI - Bristol inquiry hears closing submissions. PMID- 10678853 TI - Heart disease rising in central and eastern Europe. PMID- 10678854 TI - Clinton outlaws genetic discrimination in federal jobs. PMID- 10678855 TI - Nestle accused of breaking international code PMID- 10678856 TI - Additives may be displayed on cigarette packs. PMID- 10678857 TI - Extent and determinants of error in doctors' prognoses in terminally ill patients: prospective cohort study. AB - OBJECTIVE: To describe doctors' prognostic accuracy in terminally ill patients and to evaluate the determinants of that accuracy. DESIGN: Prospective cohort study. SETTING: Five outpatient hospice programmes in Chicago. PARTICIPANTS: 343 doctors provided survival estimates for 468 terminally ill patients at the time of hospice referral. MAIN OUTCOME MEASURES: Patients' estimated and actual survival. RESULTS: Median survival was 24 days. Only 20% (92/468) of predictions were accurate (within 33% of actual survival); 63% (295/468) were overoptimistic and 17% (81/468) were overpessimistic. Overall, doctors overestimated survival by a factor of 5.3. Few patient or doctor characteristics were associated with prognostic accuracy. Male patients were 58% less likely to have overpessimistic predictions. Non-oncology medical specialists were 326% more likely than general internists to make overpessimistic predictions. Doctors in the upper quartile of practice experience were the most accurate. As duration of doctor-patient relationship increased and time since last contact decreased, prognostic accuracy decreased. CONCLUSION: Doctors are inaccurate in their prognoses for terminally ill patients and the error is systematically optimistic. The inaccuracy is, in general, not restricted to certain kinds of doctors or patients. These phenomena may be adversely affecting the quality of care given to patients near the end of life. PMID- 10678858 TI - A memorable patient: The scars of the Jewish holocaust. PMID- 10678859 TI - Factors influencing the effect of age on prognosis in breast cancer: population based study. AB - OBJECTIVE: To investigate whether young age at diagnosis is a negative prognostic factor in primary breast cancer and how stage of disease at diagnosis and treatment influences such an association. DESIGN: Retrospective cohort study based on a population based database of patients with breast cancer containing detailed information on tumour characteristics, treatment regimens, and survival. SETTING: Denmark. SUBJECTS: 10 356 women with primary breast cancer who were less than 50 years old at diagnosis. MAIN OUTCOME MEASURES: Relative risk of dying within the first 10 years after diagnosis according to age at diagnosis after adjustment for known prognostic factors and expected mortality. RESULTS: Overall, young women with low risk disease who did not receive adjuvant treatment had a significantly increased risk of dying; risk increased with decreasing age at diagnosis (adjusted relative risk: 45-49 years (reference): 1; 40-44 years: 1.12 (95% confidence interval 0.89 to 1.40); 35-39 years: 1.40 (1.10 to 1.78); <35 years: 2.18 (1.64 to 2.89). However, no similar trend was seen in patients who received adjuvant cytotoxic treatment. The increased risk in younger women who did not receive adjuvant treatment compared with those who did remained when women were grouped according to presence of node negative disease and by tumour size. CONCLUSION: The negative prognostic effect of young age is almost exclusively seen in women diagnosed with low risk disease who did not receive adjuvant cytotoxic treatment. These results suggest that young women with breast cancer, on the basis of age alone, should be regarded as high risk patients and be given adjuvant cytotoxic treatment. PMID- 10678861 TI - Effectiveness of modified release isosorbide mononitrate affected by incorrect use. PMID- 10678860 TI - Obstructive sleep apnoea syndrome as a risk factor for hypertension: population study. AB - OBJECTIVE: To assess whether sleep apnoea syndrome is an independent risk factor for hypertension. DESIGN: Population study. SETTING: Sleep clinic in Toronto. PARTICIPANTS: 2,677 adults, aged 20-85 years, referred to the sleep clinic with suspected sleep apnoea syndrome. OUTCOME MEASURES: Medical history, demographic data, morning and evening blood pressure, and whole night polysomnography. RESULTS: Blood pressure and number of patients with hypertension increased linearly with severity of sleep apnoea, as shown by the apnoea-hypopnoea index. Multiple regression analysis of blood pressure levels of all patients not taking antihypertensives showed that apnoea was a significant predictor of both systolic and diastolic blood pressure after adjustment for age, body mass index, and sex. Multiple logistic regression showed that each additional apnoeic event per hour of sleep increased the odds of hypertension by about 1%, whereas each 10% decrease in nocturnal oxygen saturation increased the odds by 13%. CONCLUSION: Sleep apnoea syndrome is profoundly associated with hypertension independent of all relevant risk factors. PMID- 10678862 TI - Clopidogrel associated with acute arthritis. PMID- 10678863 TI - Misunderstandings in prescribing decisions in general practice: qualitative study. AB - OBJECTIVES: To identify and describe misunderstandings between patients and doctors associated with prescribing decisions in general practice. DESIGN: Qualitative study. SETTING: 20 general practices in the West Midlands and south east England. PARTICIPANTS: 20 general practitioners and 35 consulting patients. MAIN OUTCOME MEASURES: Misunderstandings between patients and doctors that have potential or actual adverse consequences for taking medicine. RESULTS: 14 categories of misunderstanding were identified relating to patient information unknown to the doctor, doctor information unknown to the patient, conflicting information, disagreement about attribution of side effects, failure of communication about doctor's decision, and relationship factors. All the misunderstandings were associated with lack of patients' participation in the consultation in terms of the voicing of expectations and preferences or the voicing of responses to doctors' decisions and actions. They were all associated with potential or actual adverse outcomes such as non-adherence to treatment. Many were based on inaccurate guesses and assumptions. In particular doctors seemed unaware of the relevance of patients' ideas about medicines for successful prescribing. CONCLUSIONS: Patients' participation in the consultation and the adverse consequences of lack of participation are important. The authors are developing an educational intervention that builds on these findings. PMID- 10678864 TI - Mortality variations as a measure of general practitioner performance: implications of the Shipman case. PMID- 10678865 TI - Before birth PMID- 10678867 TI - St Columba's case book PMID- 10678866 TI - Tropical medicine. PMID- 10678868 TI - ABC of heart failure. Management: digoxin and other inotropes, beta blockers, and antiarrhythmic and antithrombotic treatment. PMID- 10678869 TI - Internal and external morality of medicine: lessons from New Zealand. PMID- 10678870 TI - Expressing the magnitude of adverse effects in case-control studies: "the number of patients needed to be treated for one additional patient to be harmed". PMID- 10678872 TI - The royal academy exhibition PMID- 10678873 TI - Doctors write on patients' eye view of quality. Do patients want first class or economy services? PMID- 10678871 TI - "Where name and image meet"--the argument for "adrenaline". PMID- 10678874 TI - Effectiveness of rivastigmine in Alzheimer's disease. Participation in trials should be based on clinical uncertainty, not enforcement. PMID- 10678875 TI - Accuracy of perceptions of hepatitis B and C status. Injecting drug users need vaccination against hepatitis B. PMID- 10678876 TI - Acupuncture may be associated with serious adverse events. PMID- 10678877 TI - Healthcare providers in New Zealand and England could learn from each other. PMID- 10678878 TI - Bible's stance on homosexuality. Bible shows no understanding of homosexual orientation as mutually supportive and affirming. PMID- 10678879 TI - Evidence produced in evidence based medicine needs to be relevant. PMID- 10678880 TI - Fatigue and psychological distress. Statistics are improbable. PMID- 10678881 TI - Evaluation of effect of changes is essential in policymaking. PMID- 10678883 TI - Obituaries PMID- 10678882 TI - Getting HIV/AIDS accepted on the political agenda. PMID- 10678884 TI - NHS funding confused by lack of clarity PMID- 10678885 TI - The magic bullet and other medical stories PMID- 10678887 TI - ADAM interactive anatomy PMID- 10678886 TI - Assuming the risk: the mavericks, the lawyers, and the whistleblowers who beat big tobacco PMID- 10678888 TI - Netlines PMID- 10678889 TI - The stigma of schizophrenia. PMID- 10678890 TI - Tropical medicine PMID- 10678893 TI - No quick fixes PMID- 10678892 TI - Lord, protect me from my friends PMID- 10678895 TI - Chemotherapy may benefit young, low risk breast cancer patients PMID- 10678894 TI - Doctors are overoptimistic in predicting survival for patients with cancer PMID- 10678896 TI - Sleep apnoea contributes to hypertension PMID- 10678897 TI - Doctors and patients misunder- stand each other when relevant information is not exchanged PMID- 10678898 TI - Mortality data for GPs may not identify future murders PMID- 10678899 TI - Renaming adrenaline is wrong and may lead to errors PMID- 10678900 TI - Differential bacterial survival, replication, and apoptosis-inducing ability of Salmonella serovars within human and murine macrophages. AB - Salmonella serovars are associated with human diseases that range from mild gastroenteritis to host-disseminated enteric fever. Human infections by Salmonella enterica serovar Typhi can lead to typhoid fever, but this serovar does not typically cause disease in mice or other animals. In contrast, S. enterica serovar Typhimurium and S. enterica serovar Enteritidis, which are usually linked to localized gastroenteritis in humans and some animal species, elicit a systemic infection in mice. To better understand these observations, multiple strains of each of several chosen serovars of Salmonella were tested for the ability in the nonopsonized state to enter, survive, and replicate within human macrophage cells (U937 and elutriated primary cells) compared with murine macrophage cells (J774A.1 and primary peritoneal cells); in addition, death of the infected macrophages was monitored. The serovar Typhimurium strains all demonstrated enhanced survival within J774A.1 cells and murine peritoneal macrophages, compared with the significant, almost 100-fold declines in viable counts noted for serovar Typhi strains. Viable counts for serovar Enteritidis either matched the level of serovar Typhi (J774A. 1 macrophages) or were comparable to counts for serovar Typhimurium (murine peritoneal macrophages). Apoptosis was significantly higher in J774A.1 cells infected with serovar Typhimurium strain LT2 compared to serovar Typhi strain Ty2. On the other hand, serovar Typhi survived at a level up to 100-fold higher in elutriated human macrophages and 2- to 3-fold higher in U937 cells compared to the serovar Typhimurium and Enteritidis strains tested. Despite the differential multiplication of serovar Typhi during infection of U937 cells, serovar Typhi caused significantly less apoptosis than infections with serovar Typhimurium. These observations indicate variability in intramacrophage survival and host cytotoxicity among the various serovars and are the first to show differences in the apoptotic response of distinct Salmonella serovars residing in human macrophage cells. These studies suggest that nonopsonized serovar Typhimurium enters, multiplies within, and causes considerable, acute death of macrophages, leading to a highly virulent infection in mice (resulting in death within 14 days). In striking contrast, nonopsonized serovar Typhi survives silently and chronically within human macrophages, causing little cell death, which allows for intrahost dissemination and typhoid fever (low host mortality). The type of disease associated with any particular serovar of Salmonella is linked to the ability of that serovar both to persist within and to elicit damage in a specific host's macrophage cells. PMID- 10678901 TI - Reduced Th1, but not Th2, cytokine production by lymphocytes after in vivo exposure of healthy subjects to endotoxin. AB - Endotoxin (lipopolysaccharide [LPS]) tolerance is characterized by a reduced capacity of monocytes to produce proinflammatory cytokines upon restimulation in vitro. To determine whether LPS exposure induces a change in lymphocyte cytokine production and whether this results in a shift in the T-helper 1 (Th1)/Th2 balance, whole blood obtained from seven healthy subjects before and after an intravenous injection of LPS (4 ng/kg) was stimulated in vitro with the T-cell stimulus anti-CD3/CD28 or staphylococcal enterotoxin B. Whole-blood production of the Th1 cytokines gamma interferon (IFN-gamma) and interleukin-2 (IL-2) was markedly reduced at 3 and 6 h, while the production of the Th2 cytokines IL-4 and IL-5 was not influenced or was slightly increased. The IFN-gamma/IL-4 ratio was strongly decreased at 6 h. Serum obtained after LPS exposure could slightly inhibit the release of IFN-gamma but increased IL-4 production during stimulation of blood drawn from subjects not previously exposed to LPS. Normal serum also inhibited IFN-gamma production, albeit to a lesser extent. LPS exposure influences lymphocyte cytokine production, resulting in a shift toward a Th2 cytokine response, an effect that may be mediated in part by soluble factors present in serum after LPS administration in vivo. PMID- 10678902 TI - Allele substitution of the streptokinase gene reduces the nephritogenic capacity of group A streptococcal strain NZ131. AB - To investigate the role of allelic variants of streptokinase in the pathogenesis of acute poststreptococcal glomerulonephritis (APSGN), site-specific integration plasmids were constructed, which contained either the non-nephritis-associated streptokinase gene (skc5) from the group C streptococcal strain Streptococcus equisimilis H46A or the nephritis-associated streptokinase gene (ska1) from the group A streptococcal nephritogenic strain NZ131. The plasmids were introduced by electroporation and homologous recombination into the chromosome of an isogenic derivative of strain NZ131, in which the streptokinase gene had been deleted and which had thereby lost its nephritogenic capacity in a mouse model of APSGN. The introduction of a non-nephritis-associated allelic variant of streptokinase did not rescue the nephritogenic capacity of the strain. The mutant and the wild-type strains produced equivalent amounts of streptokinase. Complementation of the ska deletion derivative with the original ska allele reconstituted the nephritogenicity of wild-type NZ131. The findings support the hypothesis that the role of streptokinase in the pathogenesis of APSGN is related to the allelic variant of the protein. PMID- 10678903 TI - B cells are essential for vaccination-induced resistance to virulent Toxoplasma gondii. AB - T lymphocytes and gamma interferon (IFN-gamma) are known mediators of immune resistance to Toxoplasma gondii infection, but whether B cells also play an important role is not clear. We have investigated this issue using B-cell deficient (muMT) mice. If vaccinated with attenuated T. gondii tachyzoites, muMT mice are susceptible to a challenge intraperitoneal infection with highly virulent tachyzoites that similarly vaccinated B-cell-sufficient mice resist. Susceptibility is evidenced by increased numbers of parasites at the challenge infection site and by extensive mortality. The susceptibility of B-cell-deficient mice does not appear to be caused by deficient T-cell functions or diminished capacity of vaccinated and challenged B-cell-deficient mice to produce IFN-gamma. Administration of Toxoplasma-immune serum, but not nonimmune serum, to vaccinated B-cell-deficient mice significantly prolongs their survival after challenge with virulent tachyzoites. Vaccinated mice lacking Fc receptors or the fifth component of complement resist a challenge infection, suggesting that neither Fc-receptor dependent phagocytosis of antibody-coated tachyzoites nor antibody-dependent cellular cytotoxicity nor antibody-and-complement-dependent lysis of tachyzoites is a crucial mechanism of resistance. However, Toxoplasma-immune serum effectively inhibits the infection of host cells by tachyzoites in vitro. Together, the results support the hypothesis that B cells are required for vaccination-induced resistance to virulent tachyzoites in order to produce antibodies and that antibodies may function protectively in vivo by blocking infection of host cells by tachyzoites. PMID- 10678904 TI - Shigella flexneri 2a strain CVD 1207, with specific deletions in virG, sen, set, and guaBA, is highly attenuated in humans. AB - A phase 1 clinical trial was conducted among 35 healthy adult volunteers to evaluate the safety, immunogenicity, and shedding of different doses of CVD 1207, a live attenuated Shigella flexneri 2a vaccine candidate with specific deletion mutations in virG, sen, set, and guaBA. CVD 1207 retains the ability to invade epithelial cells but cannot effectively spread intercellularly after invasion (DeltavirG), does not produce enterotoxin (Deltasen and Deltaset), and has limited proliferation in vivo (DeltaguaBA). In a consecutive fashion, groups of three to seven subjects ingested a single oral dose of CVD 1207 at an inoculum of either 10(6), 10(7), 10(8), 10(9), or 10(10) CFU. CVD 1207 was remarkably well tolerated at inocula as high as 10(8) CFU. In comparison, one of 12 subjects who received 10(9) CFU experienced mild diarrhea and another experienced a single episode of emesis. One of five subjects who received 10(10) CFU experienced watery diarrhea and emesis. All subjects who ingested doses of 10(8) to 10(10) CFU excreted the vaccine; in 23 of 25, the duration of excretion was 1 microgram/ml in patients was recorded only for serotypes 4, 9V, 14, and 19F, which are known to be more immunogenic than serotypes 6B, 18C, and 23F. In the patient group, 70% responded to serotype 19F (Pnc 19F), 65% responded to Pnc 14 and 4, 60% responded to Pnc 9V, 55% responded to Pnc 18C, 50% responded to Pnc 23F, and 25% responded to Pnc 6B. In the control group >95% of individuals showed a titer of >1 microgram/ml to every serotype. The vaccine was tolerated well, and no major side effects have been reported. The new pneumococcal conjugate vaccine is clearly more immunogenic in previous nonresponders than is the 23-valent pneumococcal vaccine. Immunization with a pneumococcal conjugate vaccine should be considered as a strategy to protect high-risk patients. PMID- 10678958 TI - Characterization of Porphyromonas gingivalis-induced degradation of epithelial cell junctional complexes. AB - Porphyromonas gingivalis is considered among the etiological agents of human adult periodontitis. Although in vitro studies have shown that P. gingivalis has the ability to invade epithelial cell lines, its effect on the epithelial barrier junctions is not known. Immunofluorescence analysis of human gingival epithelial cells confirmed the presence of tight-junction (occludin), adherens junction (E cadherin), and cell-extracellular matrix junction (beta1-integrin) transmembrane proteins. These transmembrane proteins are expressed in Madin-Darby canine kidney (MDCK) cells. In addition, MDCK cells polarize and therefore serve as a useful in vitro model for studies on the epithelial cell barrier. Using the MDCK cell system, we examined the effect of P. gingivalis on epithelial barrier function. Exposure of the basolateral surfaces of MDCK cells to P. gingivalis (>10(9) bacteria/ml) resulted in a decrease in transepithelial resistance. Immunofluorescence microscopy demonstrated decreases in the amounts of immunoreactive occludin, E-cadherin, and beta1-integrin at specific times which were related to a disruption of cell-cell junctions in MDCK cells exposed to basolateral P. gingivalis. Disruption of cell-cell junctions was also observed upon apical exposure to bacteria; however, the effects took longer than those seen upon basolateral exposure. Cell viability was not affected by either basolateral or apical exposure to P. gingivalis. Western blot analysis demonstrated hydrolysis of occludin, E-cadherin, and beta1-integrin in lysates derived from MDCK cells exposed to P. gingivalis. Immunoprecipitated occludin and E-cadherin molecules from MDCK cell lysates were also degraded by P. gingivalis, suggesting a bacterial protease(s) capable of cleaving these epithelial junction transmembrane proteins. Collectively, these data suggest that P. gingivalis is able to invade the deeper structures of connective tissues via a paracellular pathway by degrading epithelial cell-cell junction complexes, thus allowing the spread of the bacterium. These results also indicate the importance of a critical threshold concentration of P. gingivalis to initiate epithelial barrier destruction. PMID- 10678959 TI - CpG oligodeoxynucleotides act as adjuvants for pneumococcal polysaccharide protein conjugate vaccines and enhance antipolysaccharide immunoglobulin G2a (IgG2a) and IgG3 antibodies. AB - Pneumococcal polysaccharide-protein conjugate vaccines elicit antipolysaccharide antibodies, but multiple doses are required to achieve protective antibody levels in children. In addition, the immunogenicity of experimental multivalent pneumococcal conjugate vaccines varies with different polysaccharide serotypes. One strategy to improve these vaccines is to incorporate an adjuvant to enhance their immunogenicity. Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are adjuvants that promote T-cell and T-dependent antibody responses to protein antigens, but it has been unclear whether CpG ODN can enhance polysaccharide-specific antibody responses. The present studies demonstrate significant adjuvant activity of CpG ODN for antibody responses against Streptococcus pneumoniae polysaccharide types 19F and 6B induced by conjugates of 19F and 6B with the protein carrier CRM(197). BALB/c ByJ mice were injected with 19F-CRM(197) or 6B-CRM(197) with or without CpG ODN, and sera were tested for anti-19F or anti-6B antibodies by enzyme-linked immunosorbent assay. The polysaccharide-specific antibody response to 19F-CRM(197) alone was predominantly of the immunoglobulin G1 (IgG1) and IgM isotypes, but addition of CpG ODN markedly increased geometric mean titers of total anti-19F antibody (23 fold), anti-19F IgG2a (26-fold), and anti-19F IgG3 (>246-fold). The polysaccharide-specific antibody response to 6B-CRM(197) alone consisted only of IgM, but addition of CpG ODN induced high titers of anti-6B IgG1 (>78-fold increase), anti-6B IgG2a (>54-fold increase), and anti-6B IgG3 (>3,162-fold increase). CpG ODN also increased anti-CRM(197) IgG2a and IgG3. Adjuvant effects were not observed with control non-CpG ODN. Thus, CpG ODN significantly enhance antipolysaccharide IgG responses (especially IgG2a and IgG3) induced by these glycoconjugate vaccines. PMID- 10678960 TI - Chlamydial development is adversely affected by minor changes in amino acid supply, blood plasma amino acid levels, and glucose deprivation. AB - This study has demonstrated the extreme sensitivity of Chlamydia trachomatis growing in McCoy cells to small changes in external amino acid supply. In the absence of cycloheximide, a decrease in the amino acid concentration of medium to 75% of control values was sufficient to induce the growth of enlarged chlamydial forms of reduced infectivity. Morphology became more distorted and the yield of infectious particles from inclusions declined as medium amino acid levels were further reduced. These events correlated with a general decline in intracellular amino acids, as measured by high-performance liquid chromatography, suggesting that chlamydiae require a minimum concentration of each amino acid for normal development. Cycloheximide enhanced the production of normal organisms and increased infectivity yield in media, suggesting that the drug increased the available pool of amino acids. This was supported by intracellular amino acid analyses. Aberrant forms with reduced infectivity were also induced during supply of infected cell cultures with medium containing blood plasma amino acid concentrations, supporting the proposal that nutrient levels in vivo could promote abnormal chlamydial development. Markedly abnormal forms were also observed during glucose deprivation, providing further evidence that aberrant development is a general stress-related response. PMID- 10678961 TI - Use of defined mutants to assess the role of the Campylobacter rectus S-layer in bacterium-epithelial cell interactions. AB - Campylobacter rectus is a periodontal pathogen with a 150-kDa protein on its cell surface. This protein forms a paracrystalline lattice, called the S-layer, surrounding the outer membrane of this gram-negative bacterium. To initiate a genetic analysis of the possible role of the S-layer in the initial interaction of C. rectus with host epithelial cells, C. rectus strains lacking the S-layer protein gene (crsA) were constructed by allelic exchange mutagenesis. Surprisingly, the lack of the S-layer had only a minor effect on the interaction of C. rectus with HEp-2 epithelial cells; CrsA(+) cells were 30 to 50% more adherent than were CrsA(-) bacteria. Since the host cell expression of cytokines appears to play an important role in the pathogenesis of periodontal diseases, the effect of the S-layer on the epithelial cell cytokine response was also examined by quantitative reverse transcriptase PCR and enzyme-linked immunosorbent assay. Although there were no changes in the mRNA levels for the anti-inflammatory cytokines interleukin-1 receptor agonist (IL-1ra), IL-13, and transforming growth factor beta, the expression and secretion of the proinflammatory cytokines IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) were significantly induced by both wild-type C. rectus and CrsA(-) bacteria. Interestingly, the kinetics of cytokine induction differed for the CrsA(+) and CrsA(-) bacteria. At early time points, the HEp-2 cells challenged with CrsA(-) bacteria produced higher levels of IL-6, IL-8, and TNF-alpha mRNA and protein than did cells challenged with CrsA(+) bacteria. We conclude that C. rectus may help initiate periodontitis by increasing the expression of proinflammatory cytokines and that the S-layer may temper this response to facilitate the survival of C. rectus at the site of infection. PMID- 10678962 TI - Identification of two distinct types of flagellar cap proteins, FliD, in Pseudomonas aeruginosa. AB - Binding of Pseudomonas aeruginosa strain PAK to mucin has been shown to be mediated by the flagellar cap protein, product of the fliD gene. Since the flagellar cap is very likely an exposed structure, the FliD polypeptide should be recognized by the host immune system, analogous to the recognition of dominant epitopes located in the exposed parts of the flagellin polypeptide within the assembled flagellum. In P. aeruginosa, a number of distinct flagellin variants are made, and these variable sequences presumably allow the newly infected P. aeruginosa to escape recognition by the antibody induced during a previous infection. Since similar mechanisms may direct the selection of FliD variants, we examined the extent of sequence heterogeneity among various FliD sequences among a selected group of P. aeruginosa. The results of PCR and nucleotide sequencing of the fliD region of eight different P. aeruginosa strains (laboratory strains PAK, PAO1, and PA103; clinical strains 1244, CS2, and CS32; cystic fibrosis strains CS29 and MDR) suggested that there were two distinct types of FliD in P. aeruginosa, which we named A type and B type. The results of Western blotting using the polyclonal antibodies raised against the purified FliD of A type (PAK) or B type (PAO1) further confirmed the existence of two distinct antigenic types of FliD proteins, with no cross-reactivity between the two serotypes. Further Western immunoblot analysis of the same strains using polyclonal FliC antibody showed that the strains with A-type FliD possessed a-type FliC and those with B type FliD had b-type FliC. Similar Western blot analyses of 50 more P. aeruginosa strains obtained from varied sources revealed that all strains contained either A type or B-type FliD, suggesting the existence of only two types of FliD in P. aeruginosa and indicating that fliC and fliD were coinherited. This limited diversity of FliC and FliD serotypes seems to be a unique feature of flagellar proteins. A chromosomal mutant having an insertion in the fliD gene of P. aeruginosa PAO1 was constructed. The motility defect of this mutant and a previously constructed PAK fliD mutant was better complemented with the fliD gene of the homologous types. PMID- 10678963 TI - Influence of the bcg locus on natural resistance to primary infection with the facultative intracellular bacterium Francisella tularensis in mice. AB - The implication of the Bcg locus in the control of natural resistance to infection with a live vaccine strain (LVS) of the intracellular pathogen Francisella tularensis was studied. Analysis of phenotypic expression of natural resistance and susceptibility was performed using mouse strains congenic at the Bcg locus. Comparison of the kinetics of bacterial colonization of spleen showed that B10.A.Bcg(r) mice were extremely susceptible during early phases of primary sublethal infection, while their congenic C57BL/10N [Bcg(s)] counterparts could be classified as resistant to F. tularensis LVS infection according to the 2-log lower bacterial CFU within the tissue as long as 5 days after infection. Different phenotypes of Bcg congenic mice were associated with differential expression of the cytokines tumor necrosis factor alpha, interleukin-10, and gamma interferon and production of reactive oxygen intermediates. These results strongly suggest that the Bcg locus, which is close or identical to the Nramp1 gene, controls natural resistance to infection by F. tularensis and that its effect is the opposite of that observed for other Bcg-controlled pathogens. PMID- 10678964 TI - Cellular changes and apoptosis in the spleens and peripheral blood of mice infected with blood-stage Plasmodium chabaudi chabaudi AS. AB - Infection with blood-stage Plasmodium chabaudi chabaudi AS results in splenomegaly, peripheral leukocytosis, and a major activation of the immune system. The frequencies and absolute numbers of T-cell, B-cell, and macrophage populations in spleen and peripheral blood from P. chabaudi-infected BALB/c mice were compared and found to be significantly altered during acute infection. The kinetics of the redistribution of the different cell types in spleen and peripheral blood were different, with T and B cells appearing in the blood when their frequencies and absolute numbers in the spleen were low. The frequency and absolute number of apoptotic cells in the spleen were increased during acute P. chabaudi infection and involved both T cells, B cells, and macrophages. Both Fas and Fas-ligand expression were increased in the spleen. Taken together, our data provide new information on the complex cellular interactions that take place in the immune system during blood-stage malaria infection in a mouse model. PMID- 10678965 TI - The virulence regulatory protein ToxR mediates enhanced bile resistance in Vibrio cholerae and other pathogenic Vibrio species. AB - The transmembrane regulatory protein ToxR is required for expression of virulence factors in the human diarrheal pathogen Vibrio cholerae, including cholera toxin (CT) and the toxin coregulated pilus (TCP). ToxR is necessary for transcription of the gene encoding a second regulatory protein, ToxT, which is the direct transcriptional activator of CT and TCP genes. However, ToxR, independent of ToxT, directly activates and represses transcription of the outer membrane porins OmpU and OmpT, respectively. The genes encoding TCP and CT (and including ToxT) lie on horizontally acquired genetic elements, while the toxR, ompU, and ompT genes are apparently in the ancestral Vibrio chromosome. The contribution of ToxR dependent modulation of outer membrane porins to cholera pathogenesis has remained unknown. We demonstrate that ToxR mediates enhanced bile resistance in a ToxT-independent manner. In both classical and El Tor biotypes of V. cholerae, a toxR mutant strain has a reduced minimum bactericidal concentration (MBC) of bile, the bile component deoxycholate (DC), and the anionic detergent sodium dodecyl sulfate (SDS) compared to both wild-type and toxT mutant strains. Classical and El Tor toxR mutant strains also exhibit reduced growth rates at subinhibitory concentrations of DC and SDS. Growth of either V. cholerae biotype in subinhibitory concentrations of bile or DC induces increased ToxR-dependent production of a major 38-kDa outer membrane protein, which was confirmed to be OmpU by Western blot. Measurement of transcription of a ompUp-lacZ fusion in both biotypes reveals stimulation (about two- to threefold) of ToxR-dependent ompU transcription by the presence of bile or DC, suggesting that ToxR may respond to the presence of bile. The toxR mutant strains of three additional human intestinal pathogenic Vibrio species, V. mimicus, V. fluvialis, and V. parahaemolyticus, display lower MBCs of bile, DC, and SDS and have altered outer membrane protein profiles compared to the parental wild-type strains. Our results demonstrate a conserved role for ToxR in the modulation of outer membrane proteins and bile resistance of pathogenic Vibrio species and suggest that these ToxR-dependent outer membrane proteins may mediate enhanced resistance to bile. We speculate that ToxR-mediated bile resistance was an early step in the evolution of V. cholerae as an intestinal pathogen. PMID- 10678966 TI - Listeria monocytogenes as a short-lived delivery system for the induction of type 1 cell-mediated immunity against the p36/LACK antigen of Leishmania major. AB - Listeria monocytogenes has been used as an experimental live vector for the induction of CD8-mediated immune responses in various viral and tumoral experimental models. Susceptibility of BALB/c mice to Leishmania major infection has been correlated to the preferential development of Th2 CD4 T cells through an early production of interleukin 4 (IL-4) by a restricted population of CD4 T cells which react to a single parasite antigen, LACK (stands for Leishmania homologue of receptors for activated C kinase). Experimental vaccination with LACK can redirect the differentiation of CD4(+) T cells towards the Th1 pathway if LACK is coadministrated with IL-12. As IL-12 is known to be induced by L. monocytogenes, we have tested the ability of a recombinant attenuated actA mutant L. monocytogenes strain expressing LACK to induce the development of LACK specific Th1 cells in both B10.D2 and BALB/c mice, which are resistant and susceptible to L. major, respectively. After a single injection of LACK expressing L. monocytogenes, IL-12/p40 transcripts showed a rapid burst, and peaks of gamma interferon (IFN-gamma)-secreting LACK-specific Th1 cells were detected around day 5 in the spleens and livers of mice of both strains. These primed IFN-gamma-secreting LACK-reactive T cells were not detected ex vivo after day 7 of immunization but could be recruited and detected 15 days later in the draining lymph node after an L. major footpad challenge. Although immunization of BALB/c mice with LACK-expressing L. monocytogenes did not change the course of the infection with L. major, immunized B10.D2 mice exhibited significantly smaller lesions than nonimmunized controls. Thus, our results demonstrate that, in addition of its recognized use for the induction of effector CD8 T cells, L. monocytogenes can also be used as a live recombinant vector to favor the development of potentially protective IFN-gamma-secreting Th1 CD4 T lymphocytes. PMID- 10678967 TI - Infectious CTXPhi and the vibrio pathogenicity island prophage in Vibrio mimicus: evidence for recent horizontal transfer between V. mimicus and V. cholerae. AB - Vibrio mimicus differs from Vibrio cholerae in a number of genotypic and phenotypic traits but like V. cholerae can give rise to diarrheal disease. We examined clinical isolates of V. mimicus for the presence of CTXPhi, the lysogenic filamentous bacteriophage that carries the cholera toxin genes in epidemic V. cholerae strains. Four V. mimicus isolates were found to contain complete copies of CTXPhi. Southern blot analyses revealed that V. mimicus strain PT5 contains two CTX prophages integrated at different sites within the V. mimicus genome whereas V. mimicus strains PT48, 523-80, and 9583 each contain tandemly arranged copies of CTXPhi. We detected the replicative form of CTXPhi, pCTX, in all four of these V. mimicus isolates. The CTX prophage in strain PT5 was found to produce infectious CTXPhi particles. The nucleotide sequences of CTXPhi genes orfU and zot from V. mimicus strain PT5 and V. cholerae strain N16961 were identical, indicating contemporary horizontal transfer of CTXPhi between these two species. The receptor for CTXPhi, the toxin-coregulated pilus, which is encoded by another lysogenic filamentous bacteriophage, VPIPhi, was also present in the CTXPhi-positive V. mimicus isolates. The nucleotide sequences of VPIPhi genes aldA and toxT from V. mimicus strain PT5 and V. cholerae N16961 were identical, suggesting recent horizontal transfer of this phage between V. mimicus and V. cholerae. In V. mimicus, the vibrio pathogenicity island prophage was integrated in the same chromosomal attachment site as in V. cholerae. These results suggest that V. mimicus may be a significant reservoir for both CTXPhi and VPIPhi and may play an important role in the emergence of new toxigenic V. cholerae isolates. PMID- 10678968 TI - Immunity reduces reservoir host competence of Peromyscus leucopus for Ehrlichia phagocytophila. AB - Infection with Ehrlichia phagocytophila in white-footed mice is transient and followed by a strong immune response. We investigated whether the presence of acquired immunity against E. phagocytophila precludes white-footed mice from further maintenance of this agent in nature. Mice were infected with E. phagocytophila via tick bite and challenged either 12 or 16 weeks later by Ixodes scapularis nymphs infected with the same agent. Xenodiagnostic larvae fed upon each mouse simultaneously with challenging nymphs and 1 week thereafter. Ticks were tested for the agent by PCR, and the prevalence of infection was compared to that in ticks that fed upon nonimmune control mice. Only 30% of immunized mice sustained cofeeding transmission of E. phagocytophila between simultaneously feeding infected and uninfected ticks, compared to 100% of control mice. An average of 6.3% of xenodiagnostic ticks acquired Ehrlichia from previously immunized mice when fed 1 week after the challenge, compared to 82.5% infection in the control group. Although an immune response to a single infection with E. phagocytophila in white-footed mice provided only partial protection against reinfection with the same agent, the majority of mice were rendered reservoir incompetent for at least 12 to 16 weeks. Immunity acquired by mice during I. scapularis nymphal activity in early summer may exclude a large proportion of the mouse population from maintaining E. phagocytophila during the period of larval activity later in the season. PMID- 10678969 TI - Differential regulation of CD4 lymphocyte recruitment between the upper and lower regions of the genital tract during Chlamydia trachomatis infection. AB - Genital infection with Chlamydia trachomatis results in both the local recruitment of protective immune responses and an inflammatory infiltrate that may also participate in tubal pathology. As a beginning to understanding the etiology of immune system-mediated tubal pathology, we evaluated the regional recruitment of lymphocyte subsets to different areas of the female genital tract (GT) over the course of a murine infection with the mouse pneumonitis agent of Chlamydia trachomatis (MoPn). Using flow cytometric techniques we found that the CD4 lymphocyte subset was preferentially recruited to the upper GT (oviduct and uterine horn) over the lower GT (cervical-vaginal region) throughout the course of MoPn infection. The influx of CD4 cells also correlated with the expression of endothelial cell adhesion molecules (ECAMs) and in vitro lymphocyte adherence in the upper GT. Interestingly, the expression of ECAMs in the lower GT was not maintained longer than 7 days after infection, even in the presence of viable chlamydiae. Taken together, these data suggest that regulatory mechanisms of lymphocyte recruitment differ between the upper and lower regions of the GT and may influence the clearance of chlamydiae and the development of tubal pathology. PMID- 10678970 TI - Phase 1 and phase 2 studies of Salmonella enterica serovar paratyphi A O-specific polysaccharide-tetanus toxoid conjugates in adults, teenagers, and 2- to 4-year old children in Vietnam. AB - Salmonella enterica serovar Paratyphi A O-specific polysaccharide (O-SP) was activated with 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) and bound to tetanus toxoid (TT) with adipic acid dihydrazide as a linker (SPA-TT(1)) or directly (SPA-TT(2)). In mice, these two conjugates elicited high levels of immunoglobulin G (IgG) anti-lipopolysaccharide (LPS) in serum with bactericidal activity (E. Konadu, J. Shiloach, D. A. Bryla, J. B. Robbins, and S. C. Szu, Infect. Immun. 64:2709-2715, 1996). The safety and immunogenicity of the two conjugates were then evaluated sequentially in Vietnamese adults, teenagers, and 2- to 4-year-old children. None of the vaccinees experienced significant side effects, and all had preexisting LPS antibodies. At 4 weeks after injection, there were significant increases of the geometric mean IgG and IgM anti-LPS levels in the adults and teenagers: both conjugates elicited a greater than fourfold rise in the IgG anti-LPS level in serum in >/=80% of the volunteers. SPA TT(2) elicited slightly higher, though not statistically significantly, levels of IgG anti-LPS than did SPA-TT(1) in these age groups. Accordingly, only SPA-TT(2) was evaluated in the 2- to 4-year-old children. On a random basis, one or two injections were administered 6 weeks apart to the children. No significant side effects were observed, and the levels of preexisting anti-LPS in serum were similar in children of all ages. A significant rise in the IgG anti-LPS titer was elicited by the first injection (P = 0.0001); a second injection did not elicit a booster response. Representative sera from all groups had bactericidal activity that could be adsorbed by S. enterica serovar Paratyphi A LPS. PMID- 10678971 TI - Colonization of the respiratory tract by a virulent strain of avian Escherichia coli requires carriage of a conjugative plasmid. AB - The E3 strain of E. coli was isolated in an outbreak of respiratory disease in broiler chickens, and experimental aerosol exposure of chickens to this strain induced disease similar to that seen in the field. In order to establish whether the virulent phenotype of this strain was associated with carriage of particular plasmids, four plasmid-cured derivatives, each lacking two or more of the plasmids carried by the wild-type strain, were assessed for virulence. Virulence was found to be associated with one large plasmid, pVM01. Plasmid pVM01 was marked by introduction of the transposon TnphoA, carrying kanamycin resistance, and was then cloned by transformation of E. coli strain DH5alpha. The cloned plasmid was then reintroduced by conjugation into an avirulent plasmid-cured derivative of strain E3 which lacked pVM01. The conjugant was shown to be as virulent as the wild-type strain E3, establishing that this plasmid is required for virulence following aerosol exposure. This virulence plasmid conferred expression of a hydroxamate siderophore, but not colicins, on both strain E3 and strain DH5alpha. Carriage of this plasmid was required for strain E3 to colonize the respiratory tracts of chickens but was not necessary for colonization of the gastrointestinal tract. However, the virulence plasmid did not confer virulence, or the capacity to colonize the respiratory tract, on strain DH5alpha. Thus, these studies have established that infection of chickens with E. coli strain E3 by the respiratory route is dependent on carriage of a conjugative virulence plasmid, which confers the capacity to colonize specifically the respiratory tract and which also carries genes for expression of a hydroxymate siderophore. These findings will facilitate identification of the specific genes required for virulence in these pathogens. PMID- 10678972 TI - Colonic bacteria express an ulcerative colitis pANCA-related protein epitope. AB - Bacteria are a suspected pathogenic factor in inflammatory bowel disease, but the identity of the relevant microbial species remains unresolved. The pANCA autoantibody is associated with most cases of ulcerative colitis (UC) and hence reflects an immune response associated with the disease process. This study addresses the hypothesis that pANCA identifies an antigen(s) expressed by bacteria resident in the human colonic mucosa. Libraries of colonic bacteria were generated using aerobic and anaerobic microbiologic culture conditions, and bacterial pools and clonal isolates were evaluated for cross-reactive antigens by immunoblot analysis using the pANCA monoclonal antibody Fab 5-3. Two major species of proteins immunoreactive to pANCA monoclonal antibodies were detected in bacteria from the anaerobic libraries. Colony isolates of the expressing bacteria were identified as Bacteroides caccae and Escherichia coli. Isolation and partial sequencing of the B. caccae antigen identified a 100-kDa protein without database homologous sequences. The E. coli protein was biochemically and genetically identified as the outer membrane porin OmpC. Enzyme-linked immunosorbent assay with human sera demonstrated elevated immunoglobulin G anti OmpC in UC patients compared to healthy controls. These findings demonstrate that a pANCA monoclonal antibody detects a recurrent protein epitope expressed by colonic bacteria and implicates colonic bacterial proteins as a target of the disease-associated immune response. PMID- 10678973 TI - Construction and characterization of a Salmonella enterica serovar typhimurium clone expressing a salivary adhesin of Streptococcus mutans under control of the anaerobically inducible nirB promoter. AB - Attenuated Salmonella enterica serovar Typhimurium has been used for targeted delivery of recombinant antigens to the gut-associated lymphoid tissues. One potential problem associated with this vaccine approach is the likelihood of in vivo instability of the plasmid constructs caused by constitutive hyperexpression of the heterologous immunogen. The aim of this study was to generate and characterize an expression system encoding the saliva-binding region (SBR) of Streptococcus mutans antigen I/II adhesin, either alone or linked with the mucosal adjuvant cholera toxin A2/B subunits (CTA2/B), under the control of the inducible nirB promoter. This promoter is activated in an anaerobic environment and within macrophages, which are the primary antigen-presenting cells involved in phagocytosis and processing of Salmonella. The gene encoding the chimeric SBR CTA2/B was amplified by PCR using primers containing appropriate restriction sites for subcloning into pTETnirB, which contains the nirB promoter. The resulting plasmid was introduced into serovar Typhimurium by electroporation. Production of the SBR-CTA2/B chimeric protein under anaerobic conditions was verified by enzyme-linked immunosorbent assay of whole-cell lysates on plates coated with G(M1) ganglioside and developed with antibodies to SBR. Similar procedures were followed for cloning the gene encoding SBR in serovar Typhimurium under nirB control. Anaerobic expression of SBR was confirmed by Western blotting of whole-cell lysates probed with anti-SBR antibodies. The resulting serovar Typhimurium strains were administered by either the oral or the intranasal route to mice, and colonization was assessed by microbiologic analysis of dissociated spleens, Peyer's patches (PP), and nasal tissues. High numbers of the recombinant strains persisted in PP and spleen for at least 21 days following oral challenge. A single intranasal administration of the Salmonella clones to mice also resulted in the colonization of the nasal tissues by the recombinant bacteria. Salmonellae were recovered from nasal lymphoid tissues, superficial lymph nodes, internal jugular lymph nodes, PP, and spleens of mice for at least 21 days after challenge. This study provides quantitative evidence for colonization by Salmonella strains expressing a recombinant protein under the control of the inducible nirB promoter in PP or nasal tissues following a single oral or nasal administration of the bacteria, respectively. PMID- 10678974 TI - Relative roles of pneumolysin and hydrogen peroxide from Streptococcus pneumoniae in inhibition of ependymal ciliary beat frequency. AB - Ciliated ependymal cells line the ventricular system of the brain and the cerebral aqueducts. This study characterizes the relative roles of pneumolysin and hydrogen peroxide (H(2)O(2)) in pneumococcal meningitis, using the in vitro ependymal ciliary beat frequency (CBF) as an indicator of toxicity. We have developed an ex vivo model to examine the ependymal surface of the brain slices cut from the fourth ventricle. The ependymal cells had cilia beating at a frequency of between 38 and 44Hz. D39 (wild-type) and PLN-A (pneumolysin negative) pneumococci at 10(8) CFU/ml both caused ciliary slowing. Catalase protected against PLN-A-induced ciliary slowing but afforded little protection from D39. Lysed PLN-A did not reduce CBF, whereas lysed D39 caused rapid ciliary stasis. There was no effect of catalase, penicillin, or catalase plus penicillin on the CBF. H(2)O(2) at a concentration as low as 100 microM caused ciliary stasis, and this effect was abolished by coincubation with catalase. An additive inhibition of CBF was demonstrated using a combination of both toxins. A significant inhibition of CBF at between 30 and 120 min was demonstrated with both toxins compared with either H(2)O(2) (10 microM) or pneumolysin (1 HU/ml) alone. D39 released equivalent levels of H(2)O(2) to those released by PLN-A, and these concentrations were sufficient to cause ciliary stasis. The brain slices did not produce H(2)O(2), and in the presence of 10(8) CFU of D39 or PLN-A per ml there was no detectable bacterially induced increase of H(2)O(2) release from the brain slice. Coincubation with catalase converted the H(2)O(2) produced by the pneumococci to H(2)O. Penicillin-induced lysis of bacteria dramatically reduced H(2)O(2) production. The hemolytic activity released from D39 was sufficient to cause rapid ciliary stasis, and there was no detectable release of hemolytic activity from the pneumolysin-negative PLN-A. These data demonstrate that D39 bacteria released pneumolysin, which caused rapid ciliary stasis. D39 also released H(2)O(2), which contributed to the toxicity, but this was masked by the more severe effects of pneumolysin. H(2)O(2) released from intact PLN-A was sufficient to cause rapid ciliary stasis, and catalase protected against H(2)O(2) induced cell toxicity, indicating a role for H(2)O(2) in the response. There is also a slight additive effect of pneumolysin and H(2)O(2) on ependymal toxicity; however, the precise mechanism of action and the role of these toxins in pathogenesis remain unclear. PMID- 10678975 TI - Regulation of immunoglobulin G2 production by prostaglandin E(2) and platelet activating factor. AB - Patients with localized juvenile periodontitis (LJP) have elevated levels of immunoglobulin G2 (IgG2) in their sera. This is also observed in vitro when peripheral blood leukocytes from LJP patients are stimulated with pokeweed mitogen. In previous studies, we showed that lymphocytes from subjects with no periodontitis (NP subjects) produced substantial amounts of IgG2 when they were cultured with monocytes from LJP patients (LJP monocytes). These observations indicate that monocytes or monocyte-derived mediators are positive regulators of the production of IgG2. The present study was initiated to determine if secreted factors from LJP monocytes were capable of enhancing IgG2 production and to determine if prostaglandin E2 (PGE(2)), which LJP monocytes produce at elevated levels, enhances IgG2 production. Experiments in a transwell system and with monocyte-conditioned media indicated that cell-cell contact was not necessary for LJP monocytes to augment the production of IgG2 by T and B cells from NP subjects. Moreover, the production of IgG2 was selectively induced by the addition of PGE(2) or platelet-activating factor (PAF), another lipid cytokine, which can elevate PGE(2) synthesis. Furthermore, IgG2 production was abrogated when cells were treated with indomethacin, a cyclooxygenase inhibitor that blocks the synthesis of PGE(2), or the PAF antagonists CV3988 and TEPC-15. The effects of indomethacin were completely reversed by PGE(2), indicating that this is the only prostanoid that is essential for the production of IgG2. Similarly, PGE(2) reversed the effects of a PAF antagonist, suggesting that the effects of PAF are mediated through the induction of PGE(2) synthesis. Together, these data indicate that PGE(2) and PAF are essential for the production of IgG2. PMID- 10678976 TI - Immune responses to specific antigens of Streptococcus pneumoniae and Moraxella catarrhalis in the respiratory tract. AB - Streptococcus pneumoniae and Moraxella catarrhalis are two common respiratory pathogens, colonizing as many as 54 and 72% of children, respectively, by 1 year of age. The immune responses to surface protein A of S. pneumoniae (PspA) and the high-molecular-weight outer membrane protein of M. catarrhalis (UspA) in the sera of various age groups in the general population and in the nasopharynges of 30 children monitored from birth through 1 year of age were evaluated. Immunoglobulin G (IgG) was the dominant serum antibody to PspA and UspA. Whereas the serum antibody response to PspA peaked in childhood, the antibody response to UspA peaked in adulthood. In the first 2 years of life, comparable amounts of IgM and IgG antibodies to both proteins were observed. In older persons, IgG antibodies to both antigens predominated over IgM antibodies. The levels of IgA antibody to these antigens in serum remained low during the first 2 years of life. The levels of IgM antibody to the two antigens in serum exceeded the levels of IgA antibody to the same two antigens throughout life. Although IgA was the dominant antibody to PspA and UspA in airway secretions, it was detected in a minority of the children (3 of 15 for PspA and 0 of 15 for UspA). Even the majority of the children previously colonized with these pathogens lacked antibody to them in their secretions. PMID- 10678977 TI - Molecular and evolutionary characterization of the cp32/18 family of supercoiled plasmids in Borrelia burgdorferi 297. AB - In this study, we characterized seven members of the cp32/18 family of supercoiled plasmids in Borrelia burgdorferi 297. Complete sequence analysis of a 21-kb plasmid (cp18-2) confirmed that the strain 297 plasmids are similar in overall content and organization to their B31 counterparts. Of the 31 open reading frames (ORFs) in cp18-2, only three showed sequence relatedness to proteins with known functions, and only one, a ParA/SopA ortholog, was related to nonborrelial polypeptides. Besides the lipoproteins, none of the ORFs appeared likely to encode a surface-exposed protein. Comparison with the B31 genomic sequence indicated that paralogs for most of the ORFs in cp18-2 can be identified on other genetic elements. cp18-2 was found to lack a 9- to 10-kb fragment present in the 32-kb homologs which, by extrapolation from the B31 cp32 sequences, contains at least 15 genes presumed to be unnecessary for plasmid maintenance. Sequence analysis of the lipoprotein-encoding variable loci provided evidence that recombinatorial processes within these regions may result in the acquisition of exogenous DNA. Pairwise analysis with random shuffling revealed that the multiple lipoproteins (Mlp; formerly designated 2.9 LPs) fall into two distinct homology groups which appear to have arisen by gene fusion events similar to those recently proposed to have generated the three OspE, OspF, and Elp lipoprotein families (D. R. Akins, M. J. Caimano, X. Yang, F. Cerna, M. V. Norgard, and J. D. Radolf, Infect. Immun. 67:1526-1532, 1999). Comparative analysis of the variable regions also indicated that recombination within the loci of each plasmid may occur independently. Last, comparison of variable loci revealed that the cp32/18 plasmid complements of the B31 and 297 isolates differ substantially, indicating that the two strains have been subject to divergent adaptive pressures. In addition to providing evidence for two different types of recombinatorial events involving cp32/18 plasmids, these findings underscore the need for genetic analysis of diverse borrelial isolates in order to elucidate the Lyme disease spirochete's complex parasitic strategies. PMID- 10678978 TI - Analysis of the F antigen-specific papA alleles of extraintestinal pathogenic Escherichia coli using a novel multiplex PCR-based assay. AB - Polymorphisms in PapA, the major structural subunit and antigenic determinant of P fimbriae of extraintestinal pathogenic Escherichia coli, are of considerable epidemiological, phylogenetic, and immunotherapeutic importance. However, to date, no method other than DNA sequencing has been generally available for their detection. In the present study, we developed and rigorously validated a novel PCR-based assay for the 11 recognized variants of papA and then used the new assay to assess the prevalence, phylogenetic distribution, and bacteriological associations of the papA alleles among 75 E. coli isolates from patients with urosepsis. In comparison with conventional F serotyping, the assay was extremely sensitive and specific, evidence that papA sequences are highly conserved within each of the traditionally recognized F serotypes despite the diversity observed among F types. In certain strains, the assay detected serologically occult copies of papA, of which some were shown to represent false-negative serological results and others were shown to represent the presence of nonfunctional pap fragments. Among the urosepsis isolates, the assay revealed considerable segregation of papA alleles according to O:K:H serotype, consistent with vertical transmission within clones, but with exceptions which strongly suggested horizontal transfer of papA alleles between lineages. Sequencing of papA from two strains that were papA positive by probe and PCR but F negative in the new PCR assay led to the discovery of two novel papA variants, one of which was actually more prevalent among the urosepsis isolates than were several of the known papA alleles. These findings provide novel insights into the papA alleles of extraintestinal pathogenic E. coli and indicate that the F PCR assay represents a versatile new molecular tool for epidemiological and phylogenetic investigations which should make rapid, specific detection of papA alleles available to any laboratory with PCR capability. PMID- 10678979 TI - Bacterial induction of beta interferon in mice is a function of the lipopolysaccharide component. AB - We investigated the reason for the inability of lipopolysaccharide (LPS) resistant (Lps-defective [Lps(d)]) C57BL/10ScCr mice to produce beta interferon (IFN-beta) when stimulated with bacteria. For this purpose, the IFN-beta and other macrophage cytokine responses induced by LPS and several killed gram negative and gram-positive bacteria in LPS-sensitive (Lps-normal [Lps(n)]; C57BL/10ScSn and BALB/c) and Lps(d) (C57BL/10ScCr and BALB/c/l) mice in vitro and in vivo were investigated on the mRNA and protein levels. In addition, double stranded RNA (dsRNA) was used as a nonbacterial stimulus. LPS and all gram negative bacteria employed induced IFN-beta in the Lps(n) mice but not in the Lps(d) mice. All gram-positive bacteria tested failed to induce significant amounts of IFN-beta in all four of the mouse strains used. As expected, all other cytokines tested (tumor necrosis factor alpha, interleukin 1alpha [IL-1alpha], IL 6, and IL-10) were differentially induced by gram-negative and gram-positive bacteria. Stimulation with dsRNA induced IFN-beta and all other cytokines mentioned above in all mouse strains, regardless of their LPS sensitivities. The results suggest strongly that LPS is the only bacterial component capable of inducing IFN-beta in significant amounts that are readily detectable under the conditions used in this study. Consequently, in mice, IFN-beta is inducible only by gram-negative bacteria, but not in C57BL/10ScCr or other LPS-resistant mice. PMID- 10678980 TI - Serum resistance in Haemophilus ducreyi requires outer membrane protein DsrA. AB - Haemophilus ducreyi is resistant to killing by normal serum antibody and complement. We discovered an H. ducreyi outer membrane protein required for expression of serum resistance and termed it DsrA (for "ducreyi serum resistance A"). The dsrA locus was cloned, sequenced, and mutagenized. An isogenic mutant (FX517) of parent strain 35000 was constructed and characterized, and it was found to no longer express dsrA. FX517 was at least 10-fold more serum susceptible than 35000. DsrA was expressed by all strains of H. ducreyi tested, except three naturally occurring, avirulent, serum-sensitive strains. FX517 and the three naturally occurring dsrA-nonexpressing strains were complemented in trans with a plasmid expressing dsrA. All four strains were converted to a serum resistant phenotype, including two that contained truncated lipooligosaccharide (LOS). Therefore, serum resistance in H. ducreyi does not require expression of full-length LOS but does require expression of dsrA. The dsrA locus from eight additional H. ducreyi strains was sequenced, and the deduced amino acid sequences were more than 85% identical. The major difference between the DsrA proteins was due to the presence of one, two, or three copies of the heptameric amino acid repeat NTHNINK. These repeats account for the variability in apparent molecular mass of the monomeric form of DsrA (28 to 35 kDa) observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Since DsrA is present in virulent strains, is highly conserved, and is required for serum resistance, we speculate that it may be a virulence factor and a potential vaccine candidate. PMID- 10678981 TI - Genetic resistance to experimental infection with Mycobacterium bovis in red deer (Cervus elaphus). AB - Tuberculosis (Tb) caused by Mycobacterium bovis is a worldwide threat to livestock and humans. One control strategy is to breed livestock that are more resistant to Mycobacterium bovis. In a 3-year heritability study 6 farmed red deer stags were selected from 39 on the basis of their differing responses to experimental challenge via the tonsillar sac with approximately 500 CFU of M. bovis. Two stags remained uninfected, two were moderately affected, and two developed serious spreading Tb. Seventy offspring, bred from these six stags by artificial insemination using stored semen, were similarly challenged with M. bovis. The offspring showed patterns of response to M. bovis challenge similar to those of their sires, providing evidence for a strong genetic basis to resistance to Tb, with an estimated heritability of 0.48 (standard error, 0.096; P < 0. 01). This is the first time the heritability of Tb resistance in domestic livestock has been measured. The breeding of selection lines of resistant and susceptible deer will provide an ideal model to study the mechanisms of Tb resistance in a ruminant and could provide an additional strategy for reducing the number and severity of outbreaks of Tb in farmed deer herds. Laboratory studies to identify genetic and immunological markers for resistance to Tb are under way. Preliminary studies showed no associations between NRAMP or DRB genes and resistance to Tb in deer. Patterns of immune responses seen in resistant animals suggest that both innate and acquired pathways of immunity are necessary to produce the resistant phenotype. PMID- 10678982 TI - Endotoxin-induced lung inflammation is independent of the complement membrane attack complex. AB - Several products of the activated complement system are known to modulate endothelial cell function in vitro. It has been shown that the membrane attack complex (MAC) (C5b-C9) can enhance tumor necrosis factor alpha (TNF-alpha) induced expression of P- and E-selectin and intercellular adhesion molecule type 1 in cell cultures of human umbilical vein endothelial cells. In the present study the potential role of this synegism for lung injury during endotoxin mediated septic shock in vivo was examined using a model of C6-deficient PVG (C-) (RT1(C)) rats and the congenic PVG (C+) (RT1(C)) strain. Following administration of a high (5 mg/kg) or low (0.5 mg/kg) dose of lipopolysaccharide (LPS) (Escherichia coli O55:B5), we determined the expression of cytokines, chemokines, and adhesion molecules as well as the recruitment of leukocytes in the lung. Challenge with intraperitoneal i.p. injections of LPS resulted in a strong induction of TNF-alpha, interleukin-1alpha/beta, cytokine-induced neutrophil chemoattractant, interferon-inducible protein 10, macrophage inflammatory proteins 1alpha and 2, macrophage chemotactic protein 1, and P-selectin. However, there were no significant differences between PVG (C-) and PVG (C+) rats. Immunoperoxidase staining showed a similar increase of lung infiltration by CD11b/c(+) leukocytes in both rat strains. We therefore conclude that the described synergism between TNF-alpha and the MAC of the complement system on the induction of endothelial adhesion molecules is dispensable for inflammatory processes during endotoxin-mediated septic shock in vivo. PMID- 10678984 TI - Protection against candidiasis by an immunoglobulin G3 (IgG3) monoclonal antibody specific for the same mannotriose as an IgM protective antibody. AB - We previously reported that a liposome-mannan vaccine (L-mann) of Candida albicans induces production of mouse antibodies that protect against disseminated candidiasis and vaginal infection. Immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a C. albicans cell surface beta-1,2-mannotriose, protects mice against both infections. Another IgM MAb, termed B6, which is specific for a different cell surface mannan epitope, does not protect against disseminated candidiasis. The B6.1 epitope is displayed homogeneously over the entire cell surface, compared to a patchy distribution of the B6 epitope. To determine if protection is restricted to an IgM class of antibody, we tested an IgG antibody. MAb C3.1 was obtained from L-mann-immunized mice. By results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, capture enzyme-linked immunosorbent assay, and immunodiffusion tests, MAb C3.1 is an IgG3 isotype. By epitope inhibition assays, we determined that MAb C3.1 is specific for same mannotriose as MAb B6. 1. As expected by the results of the inhibition assays, immunofluorescence microscopy showed that the C3.1 epitope is distributed on the yeast cell surface in a pattern identical to that of the B6.1 epitope. Kidney CFU and mean survival times of infected mice pretreated with MAb C3.1 indicated that the antibody enhanced resistance of mice against disseminated candidiasis. Mice in pseudoestrus that were given MAb C3.1 prior to vaginal infection developed fewer vaginal Candida CFU than control animals that received buffered saline instead of the antibody. The finding that an IgG3 antibody is protective is consistent with our hypothesis that epitope specificity and complement activation are related to the ability of an antibody to protect against candidiasis. PMID- 10678983 TI - Comparative genome analysis of the pathogenic spirochetes Borrelia burgdorferi and Treponema pallidum. AB - A comparative analysis of the predicted protein sequences encoded in the complete genomes of Borrelia burgdorferi and Treponema pallidum provides a number of insights into evolutionary trends and adaptive strategies of the two spirochetes. A measure of orthologous relationships between gene sets, termed the orthology coefficient (OC), was developed. The overall OC value for the gene sets of the two spirochetes is about 0.43, which means that less than one-half of the genes show readily detectable orthologous relationships. This emphasizes significant divergence between the two spirochetes, apparently driven by different biological niches. Different functional categories of proteins as well as different protein families show a broad distribution of OC values, from near 1 (a perfect, one-to one correspondence) to near 0. The proteins involved in core biological functions, such as genome replication and expression, typically show high OC values. In contrast, marked variability is seen among proteins that are involved in specific processes, such as nutrient transport, metabolism, gene-specific transcription regulation, signal transduction, and host response. Differences in the gene complements encoded in the two spirochete genomes suggest active adaptive evolution for their distinct niches. Comparative analysis of the spirochete genomes produced evidence of gene exchanges with other bacteria, archaea, and eukaryotic hosts that seem to have occurred at different points in the evolution of the spirochetes. Examples are presented of the use of sequence profile analysis to predict proteins that are likely to play a role in pathogenesis, including secreted proteins that contain specific protein-protein interaction domains, such as von Willebrand A, YWTD, TPR, and PR1, some of which hitherto have been reported only in eukaryotes. We tentatively reconstruct the likely evolutionary process that has led to the divergence of the two spirochete lineages; this reconstruction seems to point to an ancestral state resembling the symbiotic spirochetes found in insect guts. PMID- 10678985 TI - Influence of synthetic antiendotoxin peptides on lipopolysaccharide (LPS) recognition and LPS-induced proinflammatory cytokine responses by cells expressing membrane-bound CD14. AB - Lipopolysaccharides (LPS) are proinflammatory bacterial products implicated in the pathogenesis of gram-negative sepsis and septic shock. Polymyxin B (PMB), a cyclic, cationic peptide antibiotic, inhibits biological activities of LPS through high-affinity binding to the lipid A moiety. Small synthetic peptides have been designed to mimic the primary and secondary structures of PMB to determine structural requirements for binding and detoxification of lipid A and to assess possible therapeutic potential. The purpose of this study was to compare and contrast the endotoxin-neutralizing activities of two synthetic antiendotoxin peptides (SAEP-2 and SAEP-4), PMB, and an LPS core-specific monoclonal antibody (MAb), WN1 222-5, based on their abilities to inhibit CD14 mediated target cell uptake of fluorescein isothiocyanate (FITC)-conjugated LPS, detected by flow cytometry and confocal microscopy, and LPS-induced production of the proinflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), as measured by bioassays. PMB and SAEP-4 produced dose dependent inhibition of FITC-LPS uptake by CD14-transfected Chinese hamster ovary fibroblasts (CHO-CD14 cells) and by human peripheral blood mononuclear cells. The anti-LPS MAb, WN1 222-5, also blocked LPS uptake by these cells and synergized with PMB and SAEP-4. LPS-induced IL-6 release was inhibited by PMB, SAEP-4, and MAb WN1 222-5, and these inhibitory activities were additive or synergistic. LPS induced TNF-alpha release by PBMC was also inhibited by PMB and SAEP-4 alone and in combination with anti-LPS MAb. SAEP-2, in contrast, produced comparatively minor decrements in cellular uptake of LPS and LPS-induced cytokine responses, and did so only in the absence of serum, while a nonsense peptide exerted no discernible inhibitory effect on LPS uptake or LPS-induced cytokine expression in the presence or absence of serum. Thus, PMB and SAEP-4, like the LPS-reactive MAb, WN1 222-5, block proinflammatory activities of LPS in part by preventing LPS recognition by membrane-bound CD14-expressing target cells. Differences in peptide structure, however, like those exemplified by SAEP-2 and SAEP-4, may differentially affect the endotoxin-neutralizing potency of these peptides despite similar binding activity against lipid A, reflecting possible differences in peptide solubility or peptide regulation of intracellular signal transduction. PMID- 10678986 TI - Bacterial phosphorylcholine decreases susceptibility to the antimicrobial peptide LL-37/hCAP18 expressed in the upper respiratory tract. AB - A number of pathogens of the upper respiratory tract express an unusual prokaryotic structure, phosphorylcholine (ChoP), on their cell surface. We tested the hypothesis that ChoP, also found on host membrane lipids in the form of phosphatidylcholine, acts so as to decrease killing by antimicrobial peptides that target differences between bacterial and host membranes. In Haemophilus influenzae, ChoP is a phase-variable structure on the oligosaccharide portion of the lipopolysaccharide (LPS). There was a bactericidal effect of the peptide LL 37/hCAP18 on a nontypeable H. influenzae strain, with an increasing selection for the ChoP(+) phase as the concentration of the peptide was raised from 0 to 10 microgram/ml. Moreover, constitutive ChoP-expressing mutants of unrelated strains showed up to 1,000-fold-greater survival compared to mutants without ChoP. The effect of ChoP on resistance to killing by LL-37/hCAP18 was dependent on the salt concentration and was observed only when bacteria were grown in the presence of environmental choline, a requirement for the expression of ChoP on the LPS. Further studies established that there is transcription of the LL-37/hCAP18 gene on the epithelial surface of the human nasopharynx in situ and inducible transcription in epithelial cells derived from the upper airway. The presence of highly variable amounts of LL-37/hCAP18 in normal nasal secretions (<1.2 to >80 microgram/ml) was demonstrated with an antibody against this peptide. It was concluded that ChoP alters the bacterial cell surface so as mimic host membrane lipids and decrease killing by LL-37/hCAP18, an antimicrobial peptide that may be expressed on the mucosal surface of the nasopharynx in bactericidal concentrations. PMID- 10678988 TI - Interaction between Burkholderia pseudomallei and Acanthamoeba species results in coiling phagocytosis, endamebic bacterial survival, and escape. AB - Burkholderia pseudomallei causes melioidosis, a potentially fatal disease whose clinical outcomes include rapid-onset septicemia and relapsing and delayed-onset infections. Like other facultative intracellular bacterial pathogens, B. pseudomallei is capable of survival in human phagocytic cells, but unlike mycobacteria, Listeria monocytogenes, and Salmonella serovar Typhimurium, the species has not been reported to survive as an endosymbiont in free-living amebae. We investigated the consequences of exposing Acanthamoeba astronyxis, A. castellani, and A. polyphaga to B. pseudomallei NCTC 10276 in a series of coculture experiments. Bacterial endocytosis was observed in all three Acanthamoeba species. A more extensive range of cellular interactions including bacterial adhesion, incorporation into amebic vacuoles, and separation was observed with A. astronyxis in timed coculture experiments. Amebic trophozoites containing motile intravacuolar bacilli were found throughout 72 h of coculture. Confocal microscopy was used to confirm the intracellular location of endamebic B. pseudomallei cells. Transmission electron microscopy of coculture preparations revealed clusters of intact bacilli in membrane-lined vesicles inside the trophozoite cytoplasm; 5 x 10(2) CFU of bacteria per ml were recovered from lysed amebic trophozoites after 60 min of coculture. Demonstration of an interaction between B. pseudomallei and free-living acanthamebae in vitro raises the possibility that a similar interaction in vivo might affect environmental survival of B. pseudomallei and subsequent human exposure. Endamebic passage of B. pseudomallei warrants further investigation as a potential in vitro model of intracellular B. pseudomallei infection. PMID- 10678987 TI - Complement activation in Mycoplasma fermentans-induced mycoplasma clearance from infected cells: probing of the organism with monoclonal antibodies against M161Ag. AB - Mycoplasma fermentans, a cell wall-less prokaryote, is capable of infecting humans and has been suggested to serve as a cofactor in AIDS development. Recently, we discovered a novel lipoprotein with a molecular mass of 43 kDa originating from M. fermentans. This protein, named M161Ag, activated human complement via the alternative pathway and efficiently induced the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha, IL-6, IL-10, and IL-12 in human peripheral blood monocytes. It is likely that M161Ag of M. fermentans affects the host immune system upon mycoplasma infection. In this study, we developed monoclonal antibodies (MAbs) against M161Ag and examined the direct role of complement in M. fermentans infection using these MAbs as probes. M. fermentans was rapidly cleared from the surfaces of infected cells by human complement, but a low-grade infection persisted in human tumor cell lines. Mycoplasma particles remaining alive in host cells may cause recurrent infection, and liberated M161Ag may serve as a biological response modifier affecting both innate and acquired immunity. PMID- 10678989 TI - Roles of the surface layer proteins of Campylobacter fetus subsp. fetus in ovine abortion. AB - The role of the surface (S)-layer proteins of Campylobacter fetus subsp. fetus has been investigated using an ovine model of abortion. Wild-type strain 23D induced abortion in up to 90% of pregnant ewes challenged subcutaneously. Isolates recovered from both dams and fetuses expressed S-layer proteins with variable molecular masses. The spontaneous S-layer-negative variant, strain 23B, neither colonized nor caused abortions in pregnant ewes. A series of isogenic sapA and recA mutants, derived from 23D, also were investigated in this model. A mutant (501 [sapA recA(+)]) caused abortion in one of five challenged animals and was recovered from the placenta of a second animal. Another mutant (502 [sapA recA]) with no S-layer protein expression caused no colonization or abortions in challenged animals but caused abortion when administered intraplacentally. Mutants 600(2) and 600(4), both recA, had fixed expression of 97- and 127-kDa S layer proteins, respectively. Two of the six animals challenged with mutant 600(4) were colonized, but there were no abortions. As expected, all five strains recovered expressed a 127-kDa S-layer protein. In contrast, mutant 600(2) was recovered from the placentas of all five challenged animals and caused abortion in two. Unexpectedly, one of the 16 isolates expressed a 127-kDa rather than a 97 kDa S-layer protein. Thus, these studies indicate that S-layer proteins appear essential for colonization and/or translocation to the placenta but are not required to mediate fetal injury and that S-layer variation may occur in a recA strain. PMID- 10678990 TI - Enhanced susceptibility to subcutaneous abscess formation and persistent infection around catheters is associated with sustained interleukin-1beta levels. AB - A persistent Staphylococcus epidermidis infection in mice around a subcutaneous polyvinylpyrrolidone-grafted silicon elastomer catheter (SEpvp) but not around a conventional silicon elastomer catheter was observed. With SEpvp pericatheter tissue, protracted and exaggerated interleukin-1beta (IL-1beta) production was found. Apparently, sustained levels of IL-1beta are associated with enhanced susceptibility to biomaterial-associated S. epidermidis infection. PMID- 10678991 TI - Macropinocytosis as a mechanism of entry into primary human urethral epithelial cells by Neisseria gonorrhoeae. AB - Gonococcal entry into primary human urethral epithelial cells (HUEC) can occur by macropinocytosis. Scanning and transmission electron microscopy revealed lamellipodia surrounding gonococci, and confocal laser scanning microscopy analysis showed organisms colocalized with M(r) 70,000 fluorescein isothiocyanate labeled dextran within the cells. Phosphoinositide 3-kinase inhibitors and an actin polymerization inhibitor prevented macropinocytic entry of gonococci into HUEC. PMID- 10678992 TI - Cytotoxic cell vacuolating activity from Vibrio cholerae hemolysin. AB - A Vibrio cholerae cytotoxin, designated VcVac, was found to cause vacuolation in Vero cells. It was originally detected in the pathogenic O1 Amazonia variant of V. cholerae and later shown to be produced in environmental strains and some El Tor strains. Comparison of VcVac production in various strains suggested that hemolysin was responsible for the vacuolating phenotype. Genetic experiments established a firm correlation between vacuolation and hemolysin production. The mammalian cell vacuolating activity of the V. cholerae hemolysin is a new property of this protein and points to a previously unknown type of interaction between V. cholerae and its host. PMID- 10678993 TI - Effective preexposure tuberculosis vaccines fail to protect when they are given in an immunotherapeutic mode. AB - Two vaccine formulations previously shown to induce protective immunity in mice and prevention of long-term necrosis in guinea pigs were tested as potential immunotherapeutic vaccines in mice earlier infected by aerosol with Mycobacterium tuberculosis. Neither vaccine had any effect on the course of the infection in the lungs, but both reduced the bacterial load in the spleen. Similarly, inoculation with Mycobacterium bovis BCG had no effect whatsoever and, if given more than once, appeared to induce an increasingly severe pyogranulomatous response in the lungs of these mice. PMID- 10678994 TI - Intestinal epithelial cell apoptosis following Cryptosporidium parvum infection. AB - Cryptosporidium parvum induces moderate levels of apoptosis of cultured human intestinal epithelial cells, which are maximal at 24 h after infection. Apoptosis is further increased in C. parvum-infected cells by inhibition of NF-kappaB. C. parvum infection also attenuates epithelial apoptosis induced by strongly proapoptotic agents. The data suggest C. parvum has developed strategies to limit apoptosis in order to facilitate its growth and maturation in the early period after epithelial cell infection. PMID- 10678995 TI - Conservation and heterogeneity of vlsE among human and tick isolates of Borrelia burgdorferi. AB - The vls (variable major protein [VMP]-like sequence) locus of Borrelia burgdorferi encodes an antigenic variation system that closely resembles the VMP system of relapsing fever borreliae. To determine whether vls sequences are present consistently in low-passage, infectious isolates of B. burgdorferi, 22 blood and erythema migrans biopsy isolates from Lyme disease patients in Westchester County, New York, were examined by Southern blot and PCR analysis. Each of the strains contained a single plasmid varying in size from 21 to 38 kb that hybridized strongly with a vlsE probe based on the B. burgdorferi B31 sequence. In contrast, PCR products were obtained with only 10 of the 22 strains when primers corresponding to the 5' and 3' regions of the B31 vlsE sequence outside the variable cassette region were used. Only 2 of 16 B. burgdorferi infected tick specimens yielded detectable PCR product. Eight of 10 strains that yielded a PCR product under these conditions were type 1 (a genotype with a high rate of dissemination), according to PCR-restriction fragment length polymorphism analysis of intergenic rDNA sequences, whereas the isolates that did not yield vlsE PCR products were either type 2 or type 3. Comparison of the sequences of cloned PCR products from the patient isolates indicated a high degree of identity to the B31 sequence, with most of the differences restricted to the hypervariable regions known to undergo sequence variation. Taken together, these results both reinforce previous evidence that vls sequences are present consistently in low passage Lyme disease spirochetes and indicate that both highly conserved and heterogeneous subgroups exist with regard to vlsE sequences. PMID- 10678996 TI - T-cell epitopes in variable segments of Chlamydia trachomatis major outer membrane protein elicit serovar-specific immune responses in infected humans. AB - We previously identified 18 stimulatory Chlamydia trachomatis major outer membrane protein (MOMP) peptides containing at least 23 epitopes presented with various HLA class II allotypes. Only one peptide contained an epitope localized in a variable segment (VS2). Continued studies reported here identified a total of five VS peptides containing T-cell epitopes that are distributed among MOMPs VS1, VS2, and VS4. Only MOMP-primed T-cell cultures from subjects infected with serovar E responded to the serovar E VS peptides, while the response of such cultures to constant-segment peptides was independent of the infecting serovar. Furthermore, MOMP-primed T cells proliferated in response only to the VS peptides encoded in serovar E but not to the corresponding peptides derived from serovar F, I, or J, confirming that these responses were serovar specific. PMID- 10678997 TI - Effects of iron on extracellular and intracellular growth of Penicillium marneffei. AB - Killing of intracellular Penicillium marneffei conidia is demonstrated in gamma interferon-lipopolysaccharide-activated human THP1 and mouse J774 cells. Iron overload significantly reduces the antifungal activity of macrophages. Likewise, exogenous iron enhances and iron chelators inhibit the extracellular growth of P. marneffei. These results suggest that iron availability critically affects immunity to and the pathogenicity of P. marneffei. PMID- 10678998 TI - Stability of Borrelia burgdorferi bdr loci in vitro and in vivo. AB - The Lyme disease spirochete Borrelia burgdorferi expresses diverse subsurface yet antigenically cross-reactive Bdr protein paralogs from distinct circular- and linear-plasmid loci. We assessed the possible effects of in vitro and in vivo growth on bdr locus structure, searching for recombinational events leading to either deletions or insertions of central repeat units or novel amino- and carboxy-terminus combinations. Our data indicate that, apart from plasmid loss during in vitro cultivation, the bdr paralog loci of strain B31 are stable. This suggests that recombinatorial variation of bdr genes is not essential for persistent mammalian infection. PMID- 10678999 TI - Lipopolysaccharide entry in the damaged cornea and specific uptake by polymorphonuclear neutrophils. AB - Bacterial lipopolysaccharide (LPS) is an important agent of induction of ocular pathology following corneal injury or wearing of contaminated contact lenses. The mechanism of LPS uptake through the corneal epithelium is unclear, and the role played by inflammatory cells in this phenomenon has not been previously assessed. Fluorescein isothiocyanate-labeled LPS from Escherichia coli was deposited onto the abraded corneas of New Zealand White rabbits. Epifluorescence microscopy of living excised corneas revealed diffuse LPS staining in the epithelial and stromal layers only in the vicinity of the abrasion. In addition, specific cellular uptake of LPS was suggested by fluorescence staining of cells along the abrasion site. In a second series of experiments, an anti-CD18 polyclonal antibody was used to block infiltration of polymorphonuclear neutrophils (PMN) into the cornea. In these experiments, a diffuse distribution of fluorescent LPS was still observed along the abrasion, but the specific cellular uptake was abolished. The findings indicate that LPS enters the cornea via diffuse penetration at sites of injury and that specific cellular uptake of LPS occurs within the cornea via PMN which have migrated into the damaged tissue. PMID- 10679000 TI - Bordetella pertussis virulence factors affect phagocytosis by human neutrophils. AB - The interaction between human neutrophils and wild-type Bordetella pertussis or mutants expressing altered lipopolysaccharide or lacking virulence factors pertussis toxin, adenylate cyclase toxin, dermonecrotic toxin, filamentous hemagglutinin (FHA), pertactin, or BrkA-was examined. In the absence of antibodies, the wild-type strain and the mutants, with the exception of mutants lacking FHA, attached efficiently to neutrophils. The addition of opsonizing antibodies caused a significant reduction (approximately 50%) in attachment of the wild-type strain and most of the mutants expressing FHA, suggesting that bacterium-mediated attachment is more efficient than Fc-mediated attachment. Phagocytosis was also examined. In the absence of antibodies, about 12% of the wild-type bacteria were phagocytosed. Opsonization caused a statistically significant reduction in phagocytosis (to 3%), possibly a consequence of reduced attachment. Phagocytosis of most of the mutants was similar to that of the wild type, with the exception of the mutants lacking adenylate cyclase toxin. About 70% of the adenylate cyclase toxin mutants were phagocytosed, but only in the presence of opsonizing antibody, suggesting that Fc receptor-mediated signaling may be needed for phagocytosis. These studies indicate that FHA mediates attachment of B. pertussis to neutrophils, but adenylate cyclase toxin blocks phagocytosis. PMID- 10679001 TI - Lipopolysaccharides of Brucella abortus and Brucella melitensis induce nitric oxide synthesis in rat peritoneal macrophages. AB - Smooth lipopolysaccharide (S-LPS) and lipid A of Brucella abortus and Brucella melitensis induced the production of nitric oxide (NO) by rat adherent peritoneal cells, but they induced lower levels of production of NO than Escherichia coli LPS. The participation of the inducible isoform of NO synthase (iNOS) was confirmed by the finding of an increased expression of both iNOS mRNA and iNOS protein. These observations might help to explain (i) the acute outcome of Brucella infection in rodents, (ii) the low frequency of septic shock in human brucellosis, and (iii) the prolonged intracellular survival of Brucella in humans. PMID- 10679002 TI - Polymorphonuclear neutrophils are necessary for the recruitment of CD8(+) T cells in the liver in a pregnant mouse model of Chlamydophila abortus (Chlamydia psittaci serotype 1) infection. AB - The role of polymorphonuclear neutrophils (PMNs) in the development of the specific immune response against Chlamydophila abortus (Chlamydia psittaci serotype 1) infection was studied in a pregnant mouse model involving treatment with RB6-8C5 monoclonal antibody. PMN depletion significantly affected the immune response in the liver, in which the T-lymphocyte and F4/80(+) cell populations decreased, particularly the CD8(+) T-cell population. A Th1-like response, characterized by high levels of gamma interferon without detectable levels of interleukin 4 (IL-4) in serum, was observed in both depleted and nondepleted mice, although an increased production of IL-10 was detected in the depleted group. Our results suggest that PMNs play a very important role in the recruitment of other leukocyte populations to the inflammatory foci but have little influence in the polarization of the immune specific response toward a Th1 like response. PMID- 10679003 TI - Quantitative imaging of pre-mRNA splicing factors in living cells. PMID- 10679004 TI - Reconstitution of ATP-dependent movement of endocytic vesicles along microtubules in vitro: an oscillatory bidirectional process. AB - We have previously used the asialoglycoprotein receptor system to elucidate the pathway of hepatocytic processing of ligands such as asialoorosomucoid (ASOR). These studies suggested that endocytic vesicles bind to and travel along microtubules under the control of molecular motors such as cytoplasmic dynein. We now report reconstitution of this process in vitro with the use of a microscope assay to observe the interaction of early endocytic vesicles containing fluorescent ASOR with fluorescent microtubules. We find that ASOR-containing endosomes bind to microtubules and translocate along them in the presence of ATP. This represents the first time that mammalian endosomes containing a well characterized ligand have been directly observed to translocate on microtubules in vitro. The endosome movement does not require cytosol or exogenous motor protein, is oscillatory, and is directed toward the plus and minus ends at equal frequencies. We also observe endosomes being stretched in opposite directions along microtubules, suggesting that microtubules could provide a mechanical basis for endocytic sorting events. The movement of endosomes in vitro is consistent with the hypothesis that microtubules actively participate in the sorting and distribution of endocytic contents. PMID- 10679005 TI - Sbe2p and sbe22p, two homologous Golgi proteins involved in yeast cell wall formation. AB - The cell wall of fungal cells is important for cell integrity and cell morphogenesis and protects against harmful environmental conditions. The yeast cell wall is a complex structure consisting mainly of mannoproteins, glucan, and chitin. The molecular mechanisms by which the cell wall components are synthesized and transported to the cell surface are poorly understood. We have identified and characterized two homologous yeast proteins, Sbe2p and Sbe22p, through their suppression of a chs5 spa2 mutant strain defective in chitin synthesis and cell morphogenesis. Although sbe2 and sbe22 null mutants are viable, sbe2 sbe22 cells display several phenotypes indicative of defects in cell integrity and cell wall structure. First, sbe2 sbe22 cells display a sorbitol remediable lysis defect at 37 degrees C and are hypersensitive to SDS and calcofluor. Second, electron microscopic analysis reveals that sbe2 sbe22 cells have an aberrant cell wall structure with a reduced mannoprotein layer. Finally, immunofluorescence experiments reveal that in small-budded cells, sbe2 sbe22 mutants mislocalize Chs3p, a protein involved in chitin synthesis. In addition, sbe2 sbe22 diploids have a bud-site selection defect, displaying a random budding pattern. A Sbe2p-GFP fusion protein localizes to cytoplasmic patches, and Sbe2p cofractionates with Golgi proteins. Deletion of CHS5, which encodes a Golgi protein involved in the transport of Chs3p to the cell periphery, is lethal in combination with disruption of SBE2 and SBE22. Thus, we suggest a model in which Sbe2p and Sbe22p are involved in the transport of cell wall components from the Golgi apparatus to the cell surface periphery in a pathway independent of Chs5p. PMID- 10679006 TI - beta1 integrins regulate keratinocyte adhesion and differentiation by distinct mechanisms. AB - In keratinocytes, the beta1 integrins mediate adhesion to the extracellular matrix and also regulate the initiation of terminal differentiation. To explore the relationship between these functions, we stably infected primary human epidermal keratinocytes and an undifferentiated squamous cell carcinoma line, SCC4, with retroviruses encoding wild-type and mutant chick beta1 integrin subunits. We examined the ability of adhesion-blocking chick beta1-specific antibodies to inhibit suspension-induced terminal differentiation of primary human keratinocytes and the ability of the chick beta1 subunit to promote spontaneous differentiation of SCC4. A D154A point mutant clustered in focal adhesions but was inactive in the differentiation assays, showing that differentiation regulation required a functional ligand-binding domain. The signal transduced by beta1 integrins in normal keratinocytes was "do not differentiate" (transduced by ligand-occupied receptors) as opposed to "do differentiate" (transduced by unoccupied receptors), and the signal depended on the absolute number, rather than on the proportion, of occupied receptors. Single and double point mutations in cyto-2 and -3, the NPXY motifs, prevented focal adhesion targeting without inhibiting differentiation control. However, deletions in the proximal part of the cytoplasmic domain, affecting cyto-1, abolished the differentiation-regulatory ability of the beta1 subunit. We conclude that distinct signaling pathways are involved in beta1 integrin-mediated adhesion and differentiation control in keratinocytes. PMID- 10679007 TI - Rab7: a key to lysosome biogenesis. AB - The molecular machinery behind lysosome biogenesis and the maintenance of the perinuclear aggregate of late endocytic structures is not well understood. A likely candidate for being part of this machinery is the small GTPase Rab7, but it is unclear whether this protein is associated with lysosomes or plays any role in the regulation of the perinuclear lysosome compartment. Previously, Rab7 has mainly been implicated in transport from early to late endosomes. We have now used a new approach to analyze the role of Rab7: transient expression of Enhanced Green Fluorescent Protein (EGFP)-tagged Rab7 wt and mutant proteins in HeLa cells. EGFP-Rab7 wt was associated with late endocytic structures, mainly lysosomes, which aggregated and fused in the perinuclear region. The size of the individual lysosomes as well as the degree of perinuclear aggregation increased with the expression levels of EGFP-Rab7 wt and, more dramatically, the active EGFP-Rab7Q67L mutant. In contrast, upon expression of the dominant-negative mutants EGFP-Rab7T22N and EGFP-Rab7N125I, which localized mainly to the cytosol, the perinuclear lysosome aggregate disappeared and lysosomes, identified by colocalization of cathepsin D and lysosome-associated membrane protein-1, became dispersed throughout the cytoplasm, they were inaccessible to endocytosed molecules such as low-density lipoprotein, and their acidity was strongly reduced, as determined by decreased accumulation of the acidotropic probe LysoTracker Red. In contrast, early endosomes associated with Rab5 and the transferrin receptor, late endosomes enriched in the cation-independent mannose 6 phosphate receptor, and the trans-Golgi network, identified by its enrichment in TGN-38, were unchanged. These data demonstrate for the first time that Rab7, controlling aggregation and fusion of late endocytic structures/lysosomes, is essential for maintenance of the perinuclear lysosome compartment. PMID- 10679008 TI - Role for dynamin in late endosome dynamics and trafficking of the cation independent mannose 6-phosphate receptor. AB - It is well established that dynamin is involved in clathrin-dependent endocytosis, but relatively little is known about possible intracellular functions of this GTPase. Using confocal imaging, we found that endogenous dynamin was associated with the plasma membrane, the trans-Golgi network, and a perinuclear cluster of cation-independent mannose 6-phosphate receptor (CI-MPR) containing structures. By electron microscopy (EM), it was shown that these structures were late endosomes and that the endogenous dynamin was preferentially localized to tubulo-vesicular appendices on these late endosomes. Upon induction of the dominant-negative dynK44A mutant, confocal microscopy demonstrated a redistribution of the CI-MPR in mutant-expressing cells. Quantitative EM analysis of the ratio of CI-MPR to lysosome-associated membrane protein-1 in endosome profiles revealed a higher colocalization of the two markers in dynK44A expressing cells than in control cells. Western blot analysis showed that dynK44A expressing cells had an increased cellular procathepsin D content. Finally, EM revealed that in dynK44A-expressing cells, endosomal tubules containing CI-MPR were formed. These results are in contrast to recent reports that dynamin-2 is exclusively associated with endocytic structures at the plasma membrane. They suggest instead that endogenous dynamin also plays an important role in the molecular machinery behind the recycling of the CI-MPR from endosomes to the trans-Golgi network, and we propose that dynamin is required for the final scission of vesicles budding from endosome tubules. PMID- 10679009 TI - Nuclear pre-mRNA compartmentalization: trafficking of released transcripts to splicing factor reservoirs. AB - In the present study, the spatial organization of intron-containing pre-mRNAs of Epstein-Barr virus (EBV) genes relative to location of splicing factors is investigated. The intranuclear position of transcriptionally active EBV genes, as well as of nascent transcripts, is found to be random with respect to the speckled accumulations of splicing factors (SC35 domains) in Namalwa cells, arguing against the concept of the locus-specific organization of mRNA genes with respect to the speckles. Microclusters of splicing factors are, however, frequently superimposed on nascent transcript sites. The transcript environment is a dynamic structure consisting of both nascent and released transcripts, i.e., the track-like transcript environment. Both EBV sequences of the chromosome 1 homologue are usually associated with the track, are transcriptionally active, and exhibit in most cases a polar orientation. In contrast to nascent transcripts (in the form of spots), the association of a post-transcriptional pool of viral pre-mRNA (in the form of tracks) with speckles is not random and is further enhanced in transcriptionally silent cells when splicing factors are sequestered in enlarged accumulations. The transcript environment reflects the intranuclear transport of RNA from the sites of transcription to SC35 domains, as shown by concomitant mapping of DNA, RNA, and splicing factors. No clear vectorial intranuclear trafficking of transcripts from the site of synthesis toward the nuclear envelope for export into the cytoplasm is observed. Using Namalwa and Raji cell lines, a correlation between the level of viral gene transcription and splicing factor accumulation within the viral transcript environment has been observed. This supports a concept that the level of transcription can alter the spatial relationship among intron-containing genes, their transcripts, and speckles attributable to various levels of splicing factors recruited from splicing factor reservoirs. Electron microscopic in situ hybridization studies reveal that the released transcripts are directed toward reservoirs of splicing factors organized in clusters of interchromatin granules. Our results point to the bidirectional intranuclear movement of macromolecular complexes between intron-containing genes and splicing factor reservoirs: the recruitment of splicing factors to transcription sites and movement of released transcripts from DNA loci to reservoirs of splicing factors. PMID- 10679010 TI - Regulation of the vitellogenin receptor during Drosophila melanogaster oogenesis. AB - In many insects, development of the oocyte arrests temporarily just before vitellogenesis, the period when vitellogenins (yolk proteins) accumulate in the oocyte. Following hormonal and environmental cues, development of the oocyte resumes, and endocytosis of vitellogenins begins. An essential component of yolk uptake is the vitellogenin receptor. In this report, we describe the ovarian expression pattern and subcellular localization of the mRNA and protein encoded by the Drosophila melanogaster vitellogenin receptor gene yolkless (yl). yl RNA and protein are both expressed very early during the development of the oocyte, long before vitellogenesis begins. RNA in situ hybridization and lacZ reporter analyses show that yl RNA is synthesized by the germ line nurse cells and then transported to the oocyte. Yl protein is evenly distributed throughout the oocyte during the previtellogenic stages of oogenesis, demonstrating that the failure to take up yolk in these early stage oocyte is not due to the absence of the receptor. The transition to the vitellogenic stages is marked by the accumulation of yolk via clathrin-coated vesicles. After this transition, yolk protein receptor levels increase markedly at the cortex of the egg. Consistent with its role in yolk uptake, immunogold labeling of the receptor reveals Yl in endocytic structures at the cortex of wild-type vitellogenic oocytes. In addition, shortly after the inception of yolk uptake, we find multivesicular bodies where the yolk and receptor are distinctly partitioned. By the end of vitellogenesis, the receptor localizes predominantly to the cortex of the oocyte. However, during oogenesis in yl mutants that express full-length protein yet fail to incorporate yolk proteins, the receptor remains evenly distributed throughout the oocyte. PMID- 10679011 TI - Functional elements within the dynein microtubule-binding domain. AB - Dynein interacts with microtubules through an ATP-sensitive linkage mapped to a structurally complex region of the heavy chain following the fourth P-loop motif. Virtually nothing is known regarding how binding affinity is achieved and modulated during ATP hydrolysis. We have performed a detailed dissection of the microtubule contact site, using fragment expression, alanine substitution, and peptide competition. Our work identifies three clusters of amino acids important for the physical contact with microtubules; two of these fall within a region sharing sequence homology with MAP1B, the third in a region just downstream. Amino acid substitutions within any one of these regions can eliminate or weaken microtubule binding (KK3379, 80, E3385, K3387, K3397, KK3410,11, W3414, RKK3418 20, F3426, R3464, S3466, and K3467), suggesting that their activities are highly coordinated. A peptide that actively displaces MAP1B from microtubules perturbs dynein binding, supporting previous evidence for similar sites of interaction. We have also identified four amino acids whose substitutions affect release of the motor from the microtubule (E3413, R3444, E3460, and C3469). These suggest that nucleotide-sensitive affinity may be locally controlled at the site of contact. Our work is the first detailed description of dynein-tubulin interactions and provides a framework for understanding how affinity is achieved and modulated. PMID- 10679012 TI - Detergent-insoluble GPI-anchored proteins are apically sorted in fischer rat thyroid cells, but interference with cholesterol or sphingolipids differentially affects detergent insolubility and apical sorting. AB - In contrast to Madin-Darby canine kidney cells, Fischer rat thyroid cells deliver the majority of endogenous glycosylphosphatidyl inositol (GPI)-anchored proteins to the basolateral surface. However, we report here that the GPI proteins Placental Alkaline Phosphatase (PLAP) and Neurotrophin Receptor-Placental Alkaline Phosphatase (NTR-PLAP) are apically localized in transfected Fischer rat thyroid cells. In agreement with the "raft hypothesis," which postulates the incorporation of GPI proteins into glycosphingolipids and cholesterol-enriched rafts, we found that both of these proteins were insoluble in Triton X-100 and floated into the lighter fractions of sucrose density gradients. However, disruption of lipid rafts by removal of cholesterol did not cause surface missorting of PLAP and NTR-PLAP, and the altered surface sorting of these proteins after Fumonisin B1 treatment did not correlate with reduced levels in Triton X-100 -insoluble fractions. Furthermore, in contrast to the GPI-anchored forms of both of these proteins, the secretory and transmembrane forms (in the absence of a basolateral cytoplasmic signal) were sorted to the apical surface without association with lipid microdomains. Together, these data demonstrate that the GPI anchor is required to mediate raft association but is not sufficient to determine apical sorting. They also suggest that signals present in the ectodomain of the proteins play a major role and that lipid rafts may facilitate the recognition of these signals in the trans-Golgi network, even though they are not required for apical sorting. PMID- 10679013 TI - The puc1 cyclin regulates the G1 phase of the fission yeast cell cycle in response to cell size. AB - Eukaryotic cells coordinate cell size with cell division by regulating the length of the G1 and G2 phases of the cell cycle. In fission yeast, the length of the G1 phase depends on a precise balance between levels of positive (cig1, cig2, puc1, and cdc13 cyclins) and negative (rum1 and ste9-APC) regulators of cdc2. Early in G1, cyclin proteolysis and rum1 inhibition keep the cdc2/cyclin complexes inactive. At the end of G1, the balance is reversed and cdc2/cyclin activity down regulates both rum1 and the cyclin-degrading activity of the APC. Here we present data showing that the puc1 cyclin, a close relative of the Cln cyclins in budding yeast, plays an important role in regulating the length of G1. Fission yeast cells lacking cig1 and cig2 have a cell cycle distribution similar to that of wild-type cells, with a short G1 and a long G2. However, when the puc1(+) gene is deleted in this genetic background, the length of G1 is extended and these cells undergo S phase with a greater cell size than wild-type cells. This G1 delay is completely abolished in cells lacking rum1. Cdc2/puc1 function may be important to down-regulate the rum1 Cdk inhibitor at the end of G1. PMID- 10679014 TI - Characterization of a bone morphogenetic protein-responsive Smad-binding element. AB - Bone morphogenetic proteins (BMPs) are pleiotropic growth and differentiation factors belonging to the transforming growth factor-beta (TGF-beta) superfamily. Signals of the TGF-beta-like ligands are propagated to the nucleus through specific interaction of transmembrane serine/threonine kinase receptors and Smad proteins. GCCGnCGC has been suggested as a consensus binding sequence for Drosophila Mad regulated by a BMP-like ligand, Decapentaplegic. Smad1 is one of the mammalian Smads activated by BMPs. Here we show that Smad1 binds to this motif upon BMP stimulation in the presence of the common Smad, Smad4. The binding affinity is likely to be relatively low, because Smad1 binds to three copies of the motif weakly, but more repeats of the motif significantly enhance the binding. Heterologous reporter genes (GCCG-Lux) with multiple repeats of the motif respond to BMP stimulation but not to TGF-beta or activin. Mutational analyses reveal several bases critical for the responsiveness. A natural BMP responsive reporter, pTlx-Lux, is activated by BMP receptors in P19 cells but not in mink lung cells. In contrast, GCCG-Lux responds to BMP stimulation in both cells, suggesting that it is a universal reporter that directly detects Smad phosphorylation by BMP receptors. PMID- 10679015 TI - Conserved composition of mammalian box H/ACA and box C/D small nucleolar ribonucleoprotein particles and their interaction with the common factor Nopp140. AB - Small nucleolar ribonucleoprotein particles (snoRNPs) mainly catalyze the modification of rRNA. The two major classes of snoRNPs, box H/ACA and box C/D, function in the pseudouridylation and 2'-O-methylation, respectively, of specific nucleotides. The emerging view based on studies in yeast is that each class of snoRNPs is composed of a unique set of proteins. Here we present a characterization of mammalian snoRNPs. We show that the previously characterized NAP57 is specific for box H/ACA snoRNPs, whereas the newly identified NAP65, the rat homologue of yeast Nop5/58p, is a component of the box C/D class. Using coimmunoprecipitation experiments, we show that the nucleolar and coiled-body protein Nopp140 interacts with both classes of snoRNPs. This interaction is corroborated in vivo by the exclusive depletion of snoRNP proteins from nucleoli in cells transfected with a dominant negative Nopp140 construct. Interestingly, RNA polymerase I transcription is arrested in nucleoli depleted of snoRNPs, raising the possibility of a feedback mechanism between rRNA modification and transcription. Moreover, the Nopp140-snoRNP interaction appears to be conserved in yeast, because depletion of Srp40p, the yeast Nopp140 homologue, in a conditional lethal strain induces the loss of box H/ACA small nucleolar RNAs. We propose that Nopp140 functions as a chaperone of snoRNPs in yeast and vertebrate cells. PMID- 10679016 TI - An endosome-to-plasma membrane pathway involved in trafficking of a mutant plasma membrane ATPase in yeast. AB - The plasma membrane ATPase, encoded by PMA1, is delivered to the cell surface via the secretory pathway. Previously, we characterized a temperature-sensitive pma1 mutant in which newly synthesized Pma1-7 is not delivered to the plasma membrane but is mislocalized instead to the vacuole at 37 degrees C. Several vps mutants, which are defective in vacuolar protein sorting, suppress targeting-defective pma1 by allowing mutant Pma1 to move once again to the plasma membrane. In this study, we have analyzed trafficking in the endosomal system by monitoring the movement of Pma1-7 in vps36, vps1, and vps8 mutants. Upon induction of expression, mutant Pma1 accumulates in the prevacuolar compartment in vps36 cells. After chase, a fraction of newly synthesized Pma1-7 is delivered to the plasma membrane. In both vps1 and vps8 cells, newly synthesized mutant Pma1 appears in small punctate structures before arrival at the cell surface. Nevertheless, biosynthetic membrane traffic appears to follow different routes in vps8 and vps1: the vacuolar protein-sorting receptor Vps10p is stable in vps8 but not in vps1. Furthermore, a defect in endocytic delivery to the vacuole was revealed in vps8 (and vps36) but not vps1 by endocytosis of the bulk membrane marker FM 4-64. Moreover, in vps8 cells, there is defective down-regulation from the cell surface of the mating receptor Ste3, consistent with persistent receptor recycling from an endosomal compartment to the plasma membrane. These data support a model in which mutant Pma1 is diverted from the Golgi to the surface in vps1 cells. We hypothesize that in vps8 and vps36, in contrast to vps1, mutant Pma1 moves to the surface via endosomal intermediates, implicating an endosome-to surface traffic pathway. PMID- 10679017 TI - Roles of Hof1p, Bni1p, Bnr1p, and myo1p in cytokinesis in Saccharomyces cerevisiae. AB - Cytokinesis in Saccharomyces cerevisiae occurs by the concerted action of the actomyosin system and septum formation. Here we report on the roles of HOF1, BNI1, and BNR1 in cytokinesis, focusing on Hof1p. Deletion of HOF1 causes a temperature-sensitive defect in septum formation. A Hof1p ring forms on the mother side of the bud neck in G2/M, followed by the formation of a daughter-side ring. Around telophase, Hof1p is phosphorylated and the double rings merge into a single ring that contracts slightly and may colocalize with the actomyosin structure. Upon septum formation, Hof1p splits into two rings, disappearing upon cell separation. Hof1p localization is dependent on septins but not Myo1p. Synthetic lethality suggests that Bni1p and Myo1p belong to one functional pathway, whereas Hof1p and Bnr1p belong to another. These results suggest that Hof1p may function as an adapter linking the primary septum synthesis machinery to the actomyosin system. The formation of the actomyosin ring is not affected by bni1Delta, hof1Delta, or bnr1Delta. However, Myo1p contraction is affected by bni1Delta but not by hof1Delta or bnr1Delta. In bni1Delta cells that lack the actomyosin contraction, septum formation is often slow and asymmetric, suggesting that actomyosin contraction may provide directionality for efficient septum formation. PMID- 10679018 TI - VPS21 controls entry of endocytosed and biosynthetic proteins into the yeast prevacuolar compartment. AB - Mutations in the VPS (vacuolar protein sorting) genes of Saccharomyces cerevisiae have been used to define the trafficking steps that soluble vacuolar hydrolases take en route from the late Golgi to the vacuole. The class D VPS genes include VPS21, PEP12, and VPS45, which appear to encode components of a membrane fusion complex involved in Golgi-to-endosome transport. Vps21p is a member of the Rab family of small Ras-like GTPases and shows strong homology to the mammalian Rab5 protein, which is involved in endocytosis and the homotypic fusion of early endosomes. Although Rab5 and Vps21p appear homologous at the sequence level, it has not been clear if the functions of these two Rabs are similar. We find that Vps21p is an endosomal protein that is involved in the delivery of vacuolar and endocytosed proteins to the vacuole. Vacuolar and endocytosed proteins accumulate in distinct transport intermediates in cells that lack Vps21p function. Therefore, it appears that Vps21p is involved in two trafficking steps into the prevacuolar/late endosomal compartment. PMID- 10679019 TI - Occludin 1B, a variant of the tight junction protein occludin. AB - Occludin and claudin are the major integral membrane components of the mammalian tight junction. Although more than 11 distinct claudins have been identified, only 1 occludin transcript has been reported thus far. Therefore, we searched by reverse transcription-PCR for occludin-related sequences in Madin-Darby canine kidney (MDCK) mRNA and identified a transcript encoding an alternatively spliced form of occludin, designated occludin 1B. The occludin 1B transcript contained a 193-base pair insertion encoding a longer form of occludin with a unique N terminal sequence of 56 amino acids. Analysis of the MDCK occludin gene revealed an exon containing the 193-base pair sequence between the exons encoding the original N terminus and the distal sequence, suggesting that occludin and occludin 1B arise from alternative splicing of one transcript. To assess the expression and distribution of occludin 1B, an antibody was raised against its unique N-terminal domain. Immunolabeling of occludin 1B in MDCK cells revealed a distribution indistinguishable from that of occludin. Furthermore, occludin 1B staining at cell-to-cell contacts was also found in cultured T84 human colon carcinoma cells and in frozen sections of mouse intestine. Immunoblots of various mouse tissues revealed broad coexpression of occludin 1B with occludin. The wide epithelial distribution and the conservation across species suggests a potentially important role for occludin 1B in the structure and function of the tight junction. PMID- 10679020 TI - The role of the tethering proteins p115 and GM130 in transport through the Golgi apparatus in vivo. AB - Biochemical data have shown that COPI-coated vesicles are tethered to Golgi membranes by a complex of at least three proteins: p115, giantin, and GM130. p115 binds to giantin on the vesicles and to GM130 on the membrane. We now examine the function of this tethering complex in vivo. Microinjection of an N-terminal peptide of GM130 or overexpression of GM130 lacking this N-terminal peptide inhibits the binding of p115 to Golgi membranes. Electron microscopic analysis of single microinjected cells shows that the number of COP-sized transport vesicles in the Golgi region increases substantially, suggesting that transport vesicles continue to bud but are less able to fuse. This was corroborated by quantitative immunofluorescence analysis, which showed that the intracellular transport of the VSV-G protein was significantly inhibited. Together, these data suggest that this tethering complex increases the efficiency with which transport vesicles fuse with their target membrane. They also provide support for a model of mitotic Golgi fragmentation in which the tethering complex is disrupted by mitotic phosphorylation of GM130. PMID- 10679021 TI - The secretory pathway mediates localization of the cell polarity regulator Aip3p/Bud6p. AB - Aip3p/Bud6p is a regulator of cell and cytoskeletal polarity in Saccharomyces cerevisiae that was previously identified as an actin-interacting protein. Actin interacting protein 3 (Aip3p) localizes at the cell cortex where cytoskeleton assembly must be achieved to execute polarized cell growth, and deletion of AIP3 causes gross defects in cell and cytoskeletal polarity. We have discovered that Aip3p localization is mediated by the secretory pathway. Mutations in early- or late-acting components of the secretory apparatus lead to Aip3p mislocalization. Biochemical data show that a pool of Aip3p is associated with post-Golgi secretory vesicles. An investigation of the sequences within Aip3p necessary for Aip3p localization has identified a sequence within the N terminus of Aip3p that is sufficient for directing Aip3p localization. Replacement of the N terminus of Aip3p with a homologous region from a Schizosaccharomyces pombe protein allows for normal Aip3p localization, indicating that the secretory pathway-mediated Aip3p localization pathway is conserved. Delivery of Aip3p also requires the type V myosin motor Myo2p and its regulatory light-chain calmodulin. These data suggest that one function of calmodulin is to activate Myo2p's activity in the secretory pathway; this function is likely the polarized movement of late secretory vesicles and associated Aip3p on actin cables. PMID- 10679022 TI - Essential functions of protein tyrosine phosphatases PTP2 and PTP3 and RIM11 tyrosine phosphorylation in Saccharomyces cerevisiae meiosis and sporulation. AB - Tyrosine phosphorylation plays a central role in eukaryotic signal transduction. In yeast, MAP kinase pathways are regulated by tyrosine phosphorylation, and it has been speculated that other biochemical processes may also be regulated by tyrosine phosphorylation. Previous genetic and biochemical studies demonstrate that protein tyrosine phosphatases (PTPases) negatively regulate yeast MAP kinases. Here we report that deletion of PTP2 and PTP3 results in a sporulation defect, suggesting that tyrosine phosphorylation is involved in regulation of meiosis and sporulation. Deletion of PTP2 and PTP3 blocks cells at an early stage of sporulation before premeiotic DNA synthesis and induction of meiotic-specific genes. We observed that tyrosine phosphorylation of several proteins, including 52-, 43-, and 42-kDa proteins, was changed in ptp2Deltaptp3Delta homozygous deletion cells under sporulation conditions. The 42-kDa tyrosine-phosphorylated protein was identified as Mck1, which is a member of the GSK3 family of protein kinases and previously known to be phosphorylated on tyrosine. Mutation of MCK1 decreases sporulation efficiency, whereas mutation of RIM11, another GSK3 member, specifically abolishes sporulation; therefore, we investigated regulation of Rim11 by Tyr phosphorylation during sporulation. We demonstrated that Rim11 is phosphorylated on Tyr-199, and the Tyr phosphorylation is essential for its in vivo function, although Rim11 appears not to be directly regulated by Ptp2 and Ptp3. Biochemical characterizations indicate that tyrosine phosphorylation of Rim11 is essential for the activity of Rim11 to phosphorylate substrates. Our data demonstrate important roles of protein tyrosine phosphorylation in meiosis and sporulation PMID- 10679023 TI - Effects of I domain deletion on the function of the beta2 integrin lymphocyte function-associated antigen-1. AB - A subset of integrin alpha subunits contain an I domain, which is important for ligand binding. We have deleted the I domain from the beta2 integrin lymphocyte function-asssociated antigen-1 (LFA-1) and expressed the resulting non-I domain containing integrin (DeltaI-LFA-1) in an LFA-1-deficient T cell line. DeltaI-LFA 1 showed no recognition of LFA-1 ligands, confirming the essential role of the I domain in ligand binding. Except for I domain monoclonal antibodies (mAbs), DeltaI-LFA-1 was recognized by a panel of anti-LFA-1 mAbs similarly to wild-type LFA-1. However, DeltaI-LFA-1 had enhanced expression of seven mAb epitopes that are associated with beta2 integrin activation, suggesting that it exhibited an "active" conformation. In keeping with this characteristic, DeltaI-LFA-1 induced constitutive activation of alpha4beta1 and alpha5beta1, suggesting intracellular signaling to these integrins. This "cross-talk" was not due to an effect on beta1 integrin affinity. However, the enhanced activity was susceptible to inhibition by cytochalasin D, indicating a role for the cytoskeleton, and also correlated with clustering of beta1 integrins. Thus, removal of the I domain from LFA-1 created an integrin with the hallmarks of a constitutively active receptor mediating signals into the cell. These findings suggest a key role for the I domain in controlling integrin activity. PMID- 10679024 TI - The yeast kinesin-related protein Smy1p exerts its effects on the class V myosin Myo2p via a physical interaction. AB - We have discovered evidence for a physical interaction between a class V myosin, Myo2p, and a kinesin-related protein, Smy1p, in budding yeast. These proteins had previously been linked by genetic and colocalization studies, but we had been unable to determine the nature of their association. We now show by two-hybrid analysis that a 69-amino acid region of the Smy1p tail interacts with the globular portion of the Myo2p tail. Deletion of this myosin-binding region of Smy1p eliminates its ability to colocalize with Myo2p and to overcome the myo2-66 mutant defects, suggesting that the interaction is necessary for these functions. Further insights about the Smy1p-Myo2p interaction have come from studies of a new mutant allele, myo2-2, which causes a loss of Myo2p localization. We report that Smy1p localization is also lost in the myo2-2 mutant, demonstrating that Smy1p localization is dependent on Myo2p. We also found that overexpression of Smy1p partially restores myo2-2p localization in a myosin-binding region dependent manner. Thus, overexpression of Smy1p can overcome defects in both the head and tail domains of Myo2p (caused by the myo2-66 and myo2-2 alleles, respectively). We propose that Smy1p enhances some aspect of Myo2p function, perhaps delivery or docking of vesicles at the bud tip. PMID- 10679025 TI - Monoclonal antibodies to NTF2 inhibit nuclear protein import by preventing nuclear translocation of the GTPase Ran. AB - Nuclear transport factor 2 (NTF2) is a soluble transport protein originally identified by its ability to stimulate nuclear localization signal (NLS) dependent protein import in digitonin-permeabilized cells. NTF2 has been shown to bind nuclear pore complex proteins and the GDP form of Ran in vitro. Recently, it has been reported that NTF2 can stimulate the accumulation of Ran in digitonin permeabilized cells. Evidence that NTF2 directly mediates Ran import or that NTF2 is required to maintain the nuclear concentration of Ran in living cells has not been obtained. Here we show that cytoplasmic injection of anti-NTF2 mAbs resulted in a dramatic relocalization of Ran to the cytoplasm. This provides the first evidence that NTF2 regulates the distribution of Ran in vivo. Moreover, anti-NTF2 mAbs inhibited nuclear import of both Ran and NLS-containing protein in vitro, suggesting that NTF2 stimulates NLS-dependent protein import by driving the nuclear accumulation of Ran. We also show that biotinylated NTF2-streptavidin microinjected into the cytoplasm accumulated at the nuclear envelope, indicating that NTF2 can target a binding partner to the nuclear pore complex. Taken together, our data show that NTF2 is an essential regulator of the Ran distribution in living cells and that NTF2-mediated Ran nuclear import is required for NLS-dependent protein import. PMID- 10679026 TI - Palmitoylation of apolipoprotein B is required for proper intracellular sorting and transport of cholesteroyl esters and triglycerides. AB - Apolipoprotein B (apoB) is an essential component of chylomicrons, very low density lipoproteins, and low density lipoproteins. ApoB is a palmitoylated protein. To investigate the role of palmitoylation in lipoprotein function, a palmitoylation site was mapped to Cys-1085 and removed by mutagenesis. Secreted lipoprotein particles formed by nonpalmitoylated apoB were smaller and denser and failed to assemble a proper hydrophobic core. Indeed, the relative concentrations of nonpolar lipids were three to four times lower in lipoprotein particles containing mutant apoB compared with those containing wild-type apoB, whereas levels of polar lipids isolated from wild-type or mutant apoB lipoprotein particles appeared identical. Palmitoylation localized apoB to large vesicular structures corresponding to a subcompartment of the endoplasmic reticulum, where addition of neutral lipids was postulated to occur. In contrast, nonpalmitoylated apoB was concentrated in a dense perinuclear area corresponding to the Golgi compartment. The involvement of palmitoylation as a structural requirement for proper assembly of the hydrophobic core of the lipoprotein particle and its intracellular sorting represent novel roles for this posttranslational modification. PMID- 10679027 TI - Association of insulin receptor substrate proteins with Bcl-2 and their effects on its phosphorylation and antiapoptotic function. AB - Insulin receptor substrate (IRS) proteins are docking proteins that couple growth factor receptors to various effector molecules, including phosphoinositide-3 kinase, Grb-2, Syp, and Nck. Here we show that IRS-1 associates with the loop domain of Bcl-2 and synergistically up-regulates antiapoptotic function of Bcl-2. IRS-2 but not IRS-3 binds to Bcl-2, and IRS-1 associates with Bcl-XL but not with Bax or Bik. Overexpression of IRS-1 suppresses phosphorylation of Bcl-2 induced by stimulation with insulin, and the hypophosphorylation may lead to its enhanced antiapoptotic activity. The binding site for Bcl-2 is located on the carboxyl half-domain of IRS-1. IRS-3, which lacks the corresponding region, dominant negatively abrogates the survival effects of IRS-1 and Bcl-2. For the antiapoptotic activity of IRS-1, binding to Bcl-2 is more critical than activating phosphoinositide-3 kinase. Our results indicate that IRS proteins transmit signals from the insulin receptor to Bcl-2, thus regulating cell survival probably through regulating phosphorylation of Bcl-2. PMID- 10679028 TI - The mouse SKD1, a homologue of yeast Vps4p, is required for normal endosomal trafficking and morphology in mammalian cells. AB - The mouse SKD1 is an AAA-type ATPase homologous to the yeast Vps4p implicated in transport from endosomes to the vacuole. To elucidate a possible role of SKD1 in mammalian endocytosis, we generated a mutant SKD1, harboring a mutation (E235Q) that is equivalent to the dominant negative mutation (E233Q) in Vps4p. Overexpression of the mutant SKD1 in cultured mammalian cells caused defect in uptake of transferrin and low-density lipoprotein. This was due to loss of their receptors from the cell surface. The decrease of the surface transferrin receptor (TfR) was correlated with expression levels of the mutant protein. The mutant protein displayed a perinuclear punctate distribution in contrast to a diffuse pattern of the wild-type SKD1. TfR, the lysosomal protein lamp-1, endocytosed dextran, and epidermal growth factor but not markers for the secretory pathway were accumulated in the mutant SKD1-localized compartments. Degradation of epidermal growth factor was inhibited. Electron microscopy revealed that the compartments were exaggerated multivesicular vacuoles with numerous tubulo vesicular extensions containing TfR and endocytosed horseradish peroxidase. The early endosome antigen EEA1 was also redistributed to these aberrant membranes. Taken together, our findings suggest that SKD1 regulates morphology of endosomes and membrane traffic through them. PMID- 10679029 TI - Role of ribosome and translocon complex during folding of influenza hemagglutinin in the endoplasmic reticulum of living cells. AB - Protein folding in the living cell begins cotranslationally. To analyze how it is influenced by the ribosome and by the translocon complex during translocation into the endoplasmic reticulum, we expressed a mutant influenza hemagglutinin (a type I membrane glycoprotein) with a C-terminal extension. Analysis of the nascent chains by two-dimensional SDS-PAGE showed that ribosome attachment as such had little effect on ectodomain folding or trimer assembly. However, as long as the chains were ribosome bound and inside the translocon complex, formation of disulfides was partially suppressed, trimerization was inhibited, and the protein protected against aggregation. PMID- 10679031 TI - Progressive familial intrahepatic cholestasis: a personal perspective. AB - Progressive familial intrahepatic cholestasis (PFIC), originally described as "Byler disease" in an Amish kindred, has been distinguished from other forms of cholestatic liver disease in childhood by clinical findings, clinical-laboratory observations, and morphologic studies in biopsy, hepatectomy, and autopsy specimens. Correlation with genetic analyses has permitted both more precise definition of PFIC and distinctions within PFIC. Two types of PFIC now are recognized: PFIC-1, resulting from mutations in a gene called FIC1 (familial intrahepatic cholestasis, type 1), and PFIC-2, resulting from mutations in a gene called BSEP (bile salt export pump). Other forms of PFIC may yet be identified. The roles of FIC1 and BSEP in the secretion of bile acids into bile and in the post-secretory modification of bile are under study. PMID- 10679032 TI - Definitive classes of childhood supratentorial neuroglial tumors. The Childhood Brain Tumor Consortium. AB - Our objective in this study was to identify histologically homogenous classes of childhood supratentorial neuroglial tumors. Previously, we identified five quantitative histologic factors (differing linear combinations of 17 reliably recognized histologic features in neuroglial tumors). They account for much of the histologic variance in the 703 supratentorial tumors in the Childhood Brain Tumor Consortium (CBTC) database. In this study, we used the scores on the factors in cluster analyses and identified eight classes of neuroglial tumors. Each of these classes had significant differences in histology, allowing the separation of many of the conventional types of neuroglial tumors into two or more classes. For instance, fibrillary astrocytoma, pilocytic astrocytoma, subependymal giant cell astrocytoma, anaplastic astrocytoma, oligodendroglioma, and ependymoma were represented in two or more classes. Often these classes had statistically significant differences in survival distributions. For instance, the two classes of "anaplastic astrocytomas" have widely discrepant 5-year survival probabilities of 0.7 and 0.2. Use of the classes identified in this study ensures relatively homogeneous histologic subsets of tumors. We suggest that these classes will be useful for the selection of children for therapeutic clinical trials. PMID- 10679030 TI - Identification of novel, evolutionarily conserved Cdc42p-interacting proteins and of redundant pathways linking Cdc24p and Cdc42p to actin polarization in yeast. AB - In the yeast Saccharomyces cerevisiae, Cdc24p functions at least in part as a guanine-nucleotide-exchange factor for the Rho-family GTPase Cdc42p. A genetic screen designed to identify possible additional targets of Cdc24p instead identified two previously known genes, MSB1 and CLA4, and one novel gene, designated MSB3, all of which appear to function in the Cdc24p-Cdc42p pathway. Nonetheless, genetic evidence suggests that Cdc24p may have a function that is distinct from its Cdc42p guanine-nucleotide-exchange factor activity; in particular, overexpression of CDC42 in combination with MSB1 or a truncated CLA4 in cells depleted for Cdc24p allowed polarization of the actin cytoskeleton and polarized cell growth, but not successful cell proliferation. MSB3 has a close homologue (designated MSB4) and two more distant homologues (MDR1 and YPL249C) in S. cerevisiae and also has homologues in Schizosaccharomyces pombe, Drosophila (pollux), and humans (the oncogene tre17). Deletion of either MSB3 or MSB4 alone did not produce any obvious phenotype, and the msb3 msb4 double mutant was viable. However, the double mutant grew slowly and had a partial disorganization of the actin cytoskeleton, but not of the septins, in a fraction of cells that were larger and rounder than normal. Like Cdc42p, both Msb3p and Msb4p localized to the presumptive bud site, the bud tip, and the mother-bud neck, and this localization was Cdc42p dependent. Taken together, the data suggest that Msb3p and Msb4p may function redundantly downstream of Cdc42p, specifically in a pathway leading to actin organization. From previous work, the BNI1, GIC1, and GIC2 gene products also appear to be involved in linking Cdc42p to the actin cytoskeleton. Synthetic lethality and multicopy suppression analyses among these genes, MSB, and MSB4, suggest that the linkage is accomplished by two parallel pathways, one involving Msb3p, Msb4p, and Bni1p, and the other involving Gic1p and Gic2p. The former pathway appears to be more important in diploids and at low temperatures, whereas the latter pathway appears to be more important in haploids and at high temperatures. PMID- 10679033 TI - Congenitally malformed hearts from a population of children undergoing cardiac transplantation: comments on sequential segmental analysis and dissection. AB - Our aim is to examine the types of cardiac malformations found in a population of children undergoing cardiac transplantation, and to discuss a method for examining cardiac explants based on intrinsic morphology. We describe in detail the congenital malformations found in 65 cardiac explants acquired from a population of children over a period of 15 years. The specimens were examined and diagnosed using the method of sequential segmental analysis. The most prevalent type of cardiac malformation was severe obstruction of the left heart (29. 2%), followed by double-outlet right ventricle (15.4%), complete transposition (13.8%), hearts with left-hand ventricular topology (10.8%), ventricular septal defect(s) (9.2%), tricuspid valvar agenesis (4.6%), and tetralogy of Fallot (4.6%). These abnormalities accounted for 87.6% of the specimens studied. We also cataloged the extracardiac malformations found at autopsy in those patients who died despite the transplantation. Extracardiac malformations were identified in 10 of the 19 patients who came to autopsy. Three had heterotaxy syndrome with isomerism of the atrial appendages, one with right and two with left isomerism. Other anomalies included tracheoesophageal fistula, pulmonary sequestration, extrahepatic biliary atresia, duodenal atresia, choanal atresia, and vascular malformations.Our study shows that even the most complicated cardiac malformations can readily be diagnosed in an explanted heart using the segmental approach based on observed morphology. PMID- 10679034 TI - Bannayan-Riley-Ruvalcaba syndrome: spectrum of intestinal pathology including juvenile polyps. AB - Bannayan-Riley-Ruvalcaba syndrome (BRRS) is a disorder that includes juvenile polyposis as part of its pathologic spectrum, and it recently has been shown to share phenotypic and genotypic features with Cowden's disease. In existing literature, descriptions of intestinal pathology in patients with BRRS are relatively sparse and occasionally erroneous. We describe the intestinal pathology in multiple specimens from three children with BRRS. Examination of gastrointestinal biopsies from these children revealed predominantly colonic and rectal polyps with the histology of juvenile polyps. Additionally, two cases with clusters of ectopic ganglion cells within the lamina propria, one in a colonic polyp and one in a duodenal biopsy, and an atypical polyp were observed. Bannayan Riley-Ruvalcaba syndrome should be included in the list of differential diagnostic considerations when a child or young adult presents with a juvenile polyp, particularly if unusual histologic features such as atypical polyps or ectopic ganglion cells are encountered. PMID- 10679035 TI - Evaluation of a neuraminidase detection assay for the rapid detection of influenza A and B virus in children. AB - A prototype version of a new diagnostic assay for influenza A and B (Zstat Flutrade mark) based on detection of viral neuraminidase was evaluated and compared to culture in 196 clinical samples. Children with respiratory illnesses were prospectively evaluated at a pediatrician's office and at a large children's hospital using the neuraminidase assay and viral culture performed on respiratory secretions. Influenza virus was isolated from 51 samples and 83 were positive by the neuraminidase assay. When compared to culture the sensitivity of the assay was 96%, specificity was 77%, positive predictive value was 59%, and negative predictive value was 98%. Testing in the laboratory of pure cultures of bacteria and non-influenza viruses frequently found in the respiratory tract showed 0% cross-reactivity with the neuraminidase assay and 100% specificity for influenza virus in vitro. This new assay provided useful information for the preliminary diagnosis of influenza A and B infections and appears to be suitable for both point-of-care use in the physician's office and rapid diagnosis in a virology laboratory. The high sensitivity makes it particularly useful as a screening test for exclusion of influenza A and B infections. To confirm the diagnosis and exclude a false-positive result, as well as to determine the influenza virus type, a viral culture may be considered. PMID- 10679036 TI - Clinical and histopathological study of merosin-deficient and merosin-positive congenital muscular dystrophy. AB - The clinical features of merosin-positive congenital muscular dystrophy (CMD) and merosin-deficient CMD are well known, with those of merosin-deficient CMD being more severe. Whether the severity of histopathological findings correlates with these clinical features remains unanswered. In this study, the clinical and histopathological findings of 39 merosin-deficient and 37 merosin-positive CMD patients were compared. Merosin-deficient CMD patients were found to be younger, with earlier onset of symptoms, age of diagnosis, and a more severe clinical state (reflecting maximum motor capacity and contractures). On histopathological evaluation, endomysial fibrosis, perimysial fibrosis, and histopathological state (reflecting fibrosis, adiposis, necrosis, and variation in fiber size) were more severe in merosin-deficient CMD. There was a correlation between clinical and histopathological states only in merosin-deficient CMD. PMID- 10679037 TI - Significance of lymphoid follicles and aggregates in gastric mucosa of children. AB - This study was designed to evaluate the significance of gastric lymphoid follicles (LF) and aggregates (LA) in children with and without Helicobacter pylori (HP) infection. All 605 antrum biopsies performed during 1994 were reviewed and classified according to the presence or absence of inflammation, LF, or LA. HP was searched with a DiffQuik stain in all biopsies showing gastritis, LF, or LA. Gastritis was diagnosed in 80 biopsies (16 with LF, 18 with LA and 46 without LA or LF). Identification of HP in these biopsies was as follows: (a) cases with LF: 12/16; (b) cases with LA: 3/18; (c) cases without LF or LA: 8/46. The biopsies without gastritis had a higher frequency of LA (65/525) than of LF (2/525). HP was not identified in any case without gastritis. The presence of LF with histologic gastritis had the strongest correlation with HP (R = 0.5, p < 0.00001). LF with gastritis had a positive predictive value of 75% for HP and the absence of LF had a negative predictive value of 82.8% (sensitivity 52%; specificity 92%). LA with gastritis had no significant correlation with HP. From these results we conclude that lymphoid follicles should be distinguished from lymphoid aggregates. Lymphoid follicles can rarely be present in an otherwise normal gastric mucosa; however, they are more frequently found in cases of gastritis and are strongly associated with HP infection. Lymphoid aggregates are not significantly associated with HP infection and may be a component of the normal gastric lymphoid tissue. PMID- 10679038 TI - Novel point mutation (W184R) in neonatal type 2 Gaucher disease. AB - Gaucher disease is the most prevalent inherited sphingolipidosis and results from deficient glucocerebrosidase activity. Three clinical forms of Gaucher disease have been described: type 1, or non-neuronopathic; type 2, or acute neuronopathic; and type 3, or subacute neuronopathic. We have identified a novel mutation in a patient of Russian-British descent who died of type 2 Gaucher disease a few hours after birth. A heterozygous T --> C transition mutation in exon 6, cDNA nucleotide position 667, results in the substitution of tryptophan by arginine at amino acid residue 184 (W184R) of glucocerebrosidase. This mutation creates a new cleavage site for the restriction endonuclease Hinf1. We developed a method that utilizes Hinf1 restriction endonuclease analysis to confirm the presence of the mutation and test family members. The second mutation identified in the other glucocerebrosidase allele of the patient is mutation L444P, a severe mutation frequent in type 2 and 3 Gaucher disease. Since the patient died very shortly after birth, we postulate that the W184R/L444P genotype may result in little or no detectable glucocerebrosidase activity and thus a poor prognosis. PMID- 10679039 TI - Peripheral neuroblastic tumors: pathologic classification based on recommendations of international neuroblastoma pathology committee (Modification of shimada classification). PMID- 10679041 TI - Twenty-One Years Old-Pediatric Cardiology Coming of Age. PMID- 10679040 TI - Lysozyme immunostaining in renal tubular dysgenesis. PMID- 10679044 TI - Inhibition of germ tube formation by Candida albicans by local anesthetics: an effect related to ionic channel blockade. AB - Formation of germ tubes by Candida albicans has been assumed as a putative virulence factor. Local anesthetics (LAs), e.g., lidocaine and bupivacaine, are known to inhibit germ tube formation. The study confirmed this observation for the novel drug ropivacaine, although it was less potent than the former two drugs. Hypothesizing that the effect is due to blockading ionic channels, we exposed Candida albicans to selective calcium blockers, i.e., nifedipine and verapamil, and to a general blocker of ionic channels, i.e., lanthanum. All blockers inhibited germ tube formation. The effect was dose-dependent and pH independent. Addition of calcium reverted the effect of the blockers as well as the effect of lidocaine and ropivacaine. The study suggests that the inhibitory effect of LAs on germ tube formation by C. albicans is due to blockade of ionic channels, particularly calcium channels. Therefore, LAs can affect morphology and probably also the pathogenesis of C. albicans. PMID- 10679045 TI - Streptococcal opacity factor: a family of bifunctional proteins with lipoproteinase and fibronectin-binding activities. AB - The serum opacity factor (SOF) of Streptococcus pyogenes is a type-specific lipoproteinase of unknown biological significance. We have sequenced the sof gene and characterized the corresponding SOF protein from a strain of type M63. It was found that sof63 is related to sof22 and that, similar to SOF22 [25], SOF63 binds fibronectin. Moreover, we demonstrate opacity factor activity in a Streptococcus dysgalactiae fibronectin-binding protein FnBA that is structurally related to the SOF proteins of S. pyogenes. Sequence analysis of these three SOF proteins showed a unique periodical pattern of conserved and variable regions. The enzymatically active part of SOF63 was localized to the fragment corresponding to the entire set of conserved and variable sequences, while for fibronectin-binding a single repeat in the C terminal part of the protein was sufficient. The results show that streptococcal SOF proteins form a novel family of bifunctional proteins with lipoproteinase and fibronectin-binding activities. PMID- 10679046 TI - Use of sequence-specific antisense oligodeoxynucleotides to determine the protozoan parasite antigen recognized by nonspecific cytotoxic cells. AB - The antigen on the protozoan parasite Tetrahymena pyriformis recognized by catfish nonspecific cytotoxic cells (NCC) is a 46- to 48-kDa protein referred to as NKTag. The complete cDNA-derived amino acid sequence of NKTag has been obtained. The antigenic determinant of NKTag corresponding to the NCC binding site has been determined with synthetic peptides in target cell competition experiments. To more directly characterize the mechanism of parasite:effector cell interaction, we applied NKTag sequence-specific antisense oligodeoxynucleotides to Tetrahymena in vitro. NKTag mRNA translation by Tetrahymena was blocked by specific antisense (AS) oligodeoxynucleotides. 5'-3' sense (S) oligodeoxynucleotide sequences were synthesized corresponding to the first 17 N-terminal amino acids of NKTag (in addition to -2 untranslated codons plus the start codon). Complimentary AS oligodeoxynucleotides were likewise synthesized. To determine the optimum in vitro conditions for AS treatment, we tested parasites at various phases of their growth cycle for the effects of a single AS treatment. At 9 h post-AS treatment (during the linear phase of the growth curve), maximum reduction in membrane expression of NKTag was observed. Eighty-five percent of Tetrahymena were positive for expression of NKTag at 0 time post-AS treatment versus 13% positive at 9 h. Membrane expression of AS treated parasites returned to normal levels by 24 h post-treatment. In cold target inhibition experiments, the reduced NKTag expression by Tetrahymena at 9 h AS treatment was confirmed by observing a complete inability (compared with S treated parasites) to compete with IM-9 cells for binding with NCC. These data demonstrated a unique experimental in vitro system to define the antigen determinant on target cells responsible for recognition by cytotoxic effector cells that participate in innate immune responses. PMID- 10679047 TI - Construction of a promoter-rescue plasmid for Butyrivibrio fibrisolvens and its use in characterization of a flagellin promoter. AB - The Butyrivibrio fibrisolvens/Escherichia coli shuttle vector pBHerm has been modified to produce a plasmid (pBHE) that can be used for the identification and characterization of promoters in B. fibrisolvens. pBHE allows the insertion of a test promoter immediately upstream of a promoterless erythromycin resistance gene (ermAM). The efficacy of the pBHE plasmid in isolating and characterizing promoters was tested by inserting the flagellin gene (flaA) promoter from B. fibrisolvens OR77. Transcription of the ermAM gene from the flaA promoter was significantly higher than that observed when the ermAM gene was under the control of its own promoter. The flagelling gene of OR77 appears to be transcribed from two different promoters that produce transcripts initiating approximately 130 bp apart. Two mutant flaA promoter constructs, containing mutations in the -10 and 35 regions of either of the two putative promoter regions, showed drastic alterations in both the origin and amounts of the two transcripts produced. Mutations in either promoter affected transcription from both promoters, indicating that both regions contribute to gene expression. PMID- 10679048 TI - Mcchs1, a member of a chitin synthase gene family in Mucor circinelloides, is differentially expressed during dimorphism. AB - A complete chitin synthase gene and one chitin synthase gene fragment of the zygomycete Mucor circinelloides have been cloned and analyzed. Both genes encode zymogenic Class II chitin synthases. Hybridization analysis showed that there must exist at least another Class II chitin synthase gene in M. circinelloides highly homologous to the cloned Mcchs1 and Mcchs2. The expression of these genes during the dimorphic growing stages was analyzed. Northern hybridizations showed that Mcchs1 transcript accumulates only during the exponentially growing hyphal stage, while no expression could be detected in the yeast form. Expression of Mcchs2 could not be detected at any stage. Accumulation of Mcchs1 transcript was not influenced by visible light. The existence of a multigene chitin synthase family and the observation that Mcchs1 transcription depends upon the dimorphic stage indicate that various chitin synthase activities may have different roles in the dimorphic growth of M. circinelloides. PMID- 10679049 TI - The effect of a methanogen, Methanobrevibacter smithii, on the growth rate, organic acid production, and specific ATP activity of three predominant ruminal cellulolytic bacteria. AB - Three predominant ruminal cellulolytic organisms, Fibrobacter succinogenes S85, Ruminococcus albus 8, and R. flavefaciens FD-1, were cultured with a methanogen, Methanobrevibacter smithii. Growth rates, bacterial protein, organic acids, and methane production were measured. When grown in diculture with the methanogen, a fermentative advantage was observed with F. succinogenes S85 as seen by an increase in specific rate of ATP production and organic acid concentration. The introduction of the methanogen did not improve the growth rate, organic acid yield, or specific rate of ATP production for R. albus 8. The growth rate and amount of organic acid end products increased when R. flavefaciens FD-1 was cultured with the methanogen; however, the specific activity of ATP production did not increase. PMID- 10679050 TI - Purification and characterization of invertase from Lactobacillus reuteri CRL 1100. AB - The invertase of Lactobacillus reuteri CRL 1100 is a glycoprotein composed by a single subunit with a molecular weight of 58 kDa. The enzyme was stable below 45 degrees C over a wide pH range (4.5-7.0) with maximum activity at pH 6.0 and 37 degrees C. The invertase activity was significantly inhibited by bivalent metal ions (Ca(++), Cu(++), Cd(++), and Hg(++)), beta-mercaptoethanol, and dithiothreitol and partially improved by ethylenediaminetetraacetic acid. The enzyme was purified 32 times over the crude extract by gel filtration and ion exchange chromatography with a recovery of 17%. The K(m) and V(max) values for sucrose were 6.66 mM and 0.028 micromol/min, respectively. An invertase is purified and characterized for the first time in Lactobacillus, and it proved to be a beta-fructofuranosidase. PMID- 10679051 TI - Variations in the ability of ruminal gram-negative Prevotella species to resist monensin. AB - Gram-negative, ruminal Prevotella strains (n = 15) differed greatly in their sensitivity to the feed additive monensin. Strains that were repeatedly transferred with sublethal doses tolerated more monensin than those that were unadapted, but growth experiments indicated that the sensitivity range was as great as 2000-fold. Prevotella bryantii B(1)4 grew with monensin concentrations as high as 20 microM, but P. ruminicola H15a, D31d, 20-63, E40a, and D42f never initiated growth if monensin was greater than 0.01 microM. Washed cell preparations that were energized with glucose lost intracellular potassium when monensin was added, and potassium depletion could also be used as an index of monensin sensitivity. Adapted cells of P. bryantii B(1)4 had a half-maximal potassium depletion constant (K(d)) of 3.2 microM, but the K(d) values of P. ruminicola strains H15a, D31d, 20-63, E40a, and D42f were less than 0.04 microM. Maximal potassium depletion (K(max)) values range from 90% to 40%, and monensin adapted cells always had lower K(max) values than unadapted cells. A linear regression of log K(d)/K(max) versus percentage decrease in optical density divided by monensin concentration had an r(2) of 0.75, and this regression indicated that potassium depletion from washed cells closely correlated with growth inhibition. P. bryantii B(1)4 had a K(d)/K(max) ratio that was sevenfold greater than other Prevotella strains, and this result indicated that P. bryantii may be unusual in its ability to grow with very high concentrations of monensin. PMID- 10679052 TI - Formation of aniline as a transient metabolite during the metabolism of tetryl by a sulfate-reducing bacterial consortium. AB - A laboratory study was conducted to determine whether tetryl (2,4,6 trinitrophenylmethylnitramine) can be degraded by an anaerobic process. The results indicated that the metabolic conversion of tetryl to aniline is possible by a sulfate-reducing bacterial (SRB) consortium. This SRB consortium metabolized tetryl by co-metabolism with pyruvate as a growth substrate. For every mole of tetryl metabolized, 1 mole of aniline was produced, and the aniline was further metabolized. This metabolic conversion of tetryl is likely to be of value in the anaerobic treatment of tetryl-contaminated soil and ground water, such as found at many military ammunition sites. PMID- 10679053 TI - Biodegradability of food-associated extracellular polysaccharides. AB - Exopolysaccharides (EPSs) produced by lactic acid bacteria, which are common in fermented foods, are claimed to have various beneficial physiological effects on humans. Although the biodegradability of EPSs is important in relation to the bioactive properties, knowledge on this topic is limited. Therefore, the biodegradability of eight EPSs, six of which were produced by lactic acid bacteria, was compared with microorganisms from human feces or soil. EPS degradation was determined from the decrease in polysaccharide-sugar concentration and by high-performance size exclusion chromatography (HPSEC). Xanthan, clavan, and the EPSs produced by Streptococcus thermophilus SFi 39 and SFi 12 were readily degraded, in contrast to the EPSs produced by Lactococcus lactis ssp. cremoris B40, Lactobacillus sakei 0-1, S. thermophilus SFi20, and Lactobacillus helveticus Lh59. Clearly, the susceptibility of exopolysaccharides to biological breakdown can differ greatly, implying that the physiological effects of these compounds may also vary a lot. PMID- 10679054 TI - The crystal proteins from Bacillus thuringiensis subsp. thompsoni display a synergistic activity against the codling moth, Cydia pomonella. AB - Crystal proteins from Bacillus thuringiensis subsp. thompsoni strain HnC are active against the codling moth, Cydia pomonella, a major pest of orchards. Inclusion bodies purified from strain HnC displayed an LC(50) of 3.34 x 10(-3) microgram/microliter. HnC-purified crystals were tenfold more active than Cry2Aa and Cry1Aa toxins, and 100-fold more toxic than Cry1Ab. The 34-kDa and 40-kDa proteins contained in HnC inclusion bodies were shown to act synergistically. The toxicity of crystal proteins produced by the recombinant B. thuringiensis strain BT-OP expressing the full-length native operon was about tenfold higher than that of the 34-kDa protein. When the gene encoding the non-insecticidal 40-kDa protein, which is not active, was introduced into the recombinant strain producing only the 34-kDa protein, the toxicity was raised tenfold and was similar to that of the strain BT-OP. PMID- 10679055 TI - Occurrence of Bacillus thuringiensis on cured tobacco leaves. AB - A worldwide survey was conducted to evaluate the frequency and distribution of Bacillus thuringiensis populations on cured tobacco leaves during post-harvest storage. In total, 133 tobacco samples of different types and origins were analyzed. Nine percent of the samples showed the presence of B. thuringiensis, and 24 B. thuringiensis strains were isolated and characterized. The majority of the isolates produced bipyramidal crystals, and three fourths of them showed a second type of crystal protein (cuboidal or heterogeneous crystals). Only three isolates showed the rhomboidal crystal morphology characteristic of the anti coleopteran B. thuringiensis subsp. tenebrionis. PCR analysis with primers specific for cry1 and cry3 genes revealed eight distinct cry gene profiles. The results of this study indicate that B. thuringiensis is naturally present at low frequency on the phylloplane of cured tobacco leaves and that its distribution is worldwide. PMID- 10679056 TI - Influence of sulfate-reducing bacteria on outdoor hydrogen production by photosynthetic bacterium with seawater. AB - The application of seawater for bacterial fermentative production is a cost effective technology. Hydrogen production by marine photosynthetic bacterium with seawater failed to continue after more than 10 days, and was accompanied by the formation of hydrogen sulfide and a change in culture color from red to black. However, substrate consumption in the blackish culture was comparable to that in a hydrogen-producing culture. A decrease in hydrogen production occurred upon the addition of sodium sulfide at concentrations of 1.5 mM or higher. PCR analysis targeted at the 16S rDNA sequence selective for sulfate-reducing bacteria revealed the existence of sulfate-reducing bacteria in inoculation cultures of the phototrophic bacterium and medium for hydrogen production. Hence, the high sulfate concentration of seawater, the low oxidation-reduction potential under hydrogen-producing conditions, and the presence of electron donors such as acetate might promote the metabolic activities of sulfate-reducing bacteria, resulting in the deterioration of hydrogen production with seawater. PMID- 10679058 TI - Cutting edge: lipoxin (LX) A4 and aspirin-triggered 15-epi-LXA4 block allergen induced eosinophil trafficking. AB - Tissue eosinophilia prevention represents one of the primary targets to new anti allergic therapies. As lipoxin A4 (LXA4) and aspirin-triggered 15-epi-LXA4 (ATL) are emerging as endogenous "stop signals" produced in distinct pathologies including some eosinophil-related pulmonary disorders, we evaluated the impact of in situ LXA4/ATL metabolically stable analogues on allergen-induced eosinophilic pleurisy in sensitized rats. LXA4/ATL analogues dramatically blocked allergic pleural eosinophil influx, while concurrently increasing circulating eosinophilia, inhibiting the earlier edema and neutrophilia associated with allergic reaction. The mechanisms underlying this LXA4/ATL-driven allergic eosinophilia blockade was independent of mast cell degranulation and involved LXA4/ATL inhibition of both IL-5 and eotaxin generation, as well as platelet activating factor action. These findings reveal LXA4/ATL as a novel class of endogenous anti-allergic mediators, capable of preventing local eosinophilia. PMID- 10679059 TI - Cutting edge: functional role for proline-rich tyrosine kinase 2 in NK cell mediated natural cytotoxicity. AB - Protein tyrosine kinase activation is one of the first biochemical events in the signaling pathway leading to activation of NK cell cytolytic machinery. Here we investigated whether proline-rich tyrosine kinase 2 (Pyk2), the nonreceptor protein tyrosine kinase belonging to the focal adhesion kinase family, could play a role in NK cell-mediated cytotoxicity. Our results demonstrate that binding of NK cells to sensitive target cells or ligation of beta2 integrins results in a rapid induction of Pyk2 phosphorylation and activation. By contrast, no detectable Pyk2 tyrosine phosphorylation is found upon CD16 stimulation mediated by either mAb or interaction with Ab-coated P815 cells. A functional role for Pyk2 in natural but not Ab-mediated cytotoxicity was demonstrated by the use of recombinant vaccinia viruses encoding the kinase dead mutant of Pyk2. Finally, we provide evidence that Pyk2 is involved in the beta2 integrin-triggered extracellular signal-regulated kinase activation, supporting the hypothesis that Pyk2 plays a role in the natural cytotoxicity by controlling extracellular signal regulated kinase activation. PMID- 10679060 TI - Cutting edge: differential production of prostaglandin D2 by human helper T cell subsets. AB - Several effector molecules, including cytokines, are differentially produced by Th1 and Th2 cells. We used a gene expression screen method to identify a gene encoding hematopoietic PG D synthase (hPGDS) which was preferentially expressed in human Th2 but not Th1 clones. Studies with anti-hPGDS mAbs confirmed the Th2 dominated expression of hPGDS protein. Upon stimulation with anti-CD3 plus anti CD28 mAbs, coordinated cyclooxygenase-2 expression and PGD2 production were induced in Th2 lines. hPGDS expression was also observed in a small population (<1.0%) of peripheral blood CD4+ lymphocytes from healthy adults. Most hPGDS expressing CD4+ lymphocytes showed a typical Th2-type cytokine pattern. Our results suggest that, at the sites of Ag presentation, at least part of the Th2 cell population produces PGD2, which may be involved in various aspects of Th2 related immune responses similar to mast cells. PMID- 10679061 TI - Cutting edge: complement-activating complex of ficolin and mannose-binding lectin associated serine protease. AB - Both ficolins and mannose-binding lectin (MBL) are lectins characterized by the presence of collagen-like and carbohydrate-binding domains in a subunit, although their carbohydrate-binding moieties are quite different. A fibrinogen-like domain is in ficolins, and a carbohydrate recognition domain is in MBL. On binding to pathogens, human MBL activates the complement system via the lectin pathway in association with two types of MBL-associated serine proteases (MASP), MASP-1 and MASP-2 and its truncated form, small MBL-associated protein (sMAP, also called MAp19). We report here that ficolin/P35, a human serum ficolin, was found to copurify with MASPs and sMAP. MASPs that were complexed with ficolin/P35 exhibited proteolytic activities against complement components C4, C2, and C3. The ficolin/P35-MASPs-sMAP complex that was bound to Salmonella typhimurium activated complement. These findings indicate that ficolin/P35 is a second collagenous lectin capable of activating the lectin pathway and thus plays a role in innate immunity. PMID- 10679062 TI - TGF-beta 1 reciprocally controls chemotaxis of human peripheral blood monocyte derived dendritic cells via chemokine receptors. AB - We examined the effect of TGF-beta 1 on the chemotactic migratory ability of human monocyte-derived dendritic cells (DCs). Treatment of immature DCs with TGF beta 1 resulted in increased expressions of CCR-1, CCR-3, CCR-5, CCR-6, and CXC chemokine receptor-4 (CXCR-4), which were concomitant with enhanced chemotactic migratory responses to their ligands, RANTES (for CCR-1, CCR-3, and CCR-5), macrophage-inflammatory protein-3 alpha (MIP-3 alpha) (for CCR-6), or stromal cell-derived growth factor-1 alpha (for CXCR-4). Ligation by TNF-alpha resulted in down-modulation of cell surface expressions of CCR-1, CCR-3, CCR-5, CCR-6, and CXCR-4, and the chemotaxis for RANTES, MIP-3 alpha, and stromal cell-derived growth factor-1 alpha, whereas this stimulation up-regulated the expression of CCR-7 and the chemotactic ability for MIP-3beta. Stimulation of mature DCs with TGF-beta 1 also enhanced TNF-alpha-induced down-regulation of the expressions of CCR-1, CCR-3, CCR-5, CCR-6, and CXCR-4, and chemotaxis to their respective ligands, while this stimulation suppressed TNF-alpha-induced expression of CCR-7 and chemotactic migratory ability to MIP-3 beta. Our findings suggest that TGF beta 1 reversibly regulates chemotaxis of DCs via regulation of chemokine receptor expression. PMID- 10679063 TI - IFN-alpha 2b reduces IL-2 production and IL-2 receptor function in primary CD4+ T cells. AB - Initially described as an antiviral cytokine, IFN-alpha has been subsequently shown to affect several cellular functions, including cellular differentiation and proliferation. For these reasons, IFN-alpha is currently used in clinical practice for the treatment of viral infections and malignancies. In this manuscript, we show two novel mechanisms concomitantly responsible for the antiproliferative effect of IFN-alpha. First, long-term treatment with IFN-alpha of primary CD4+ T cells reduced surface expression of CD3 and CD28. These events resulted in decreased phosphorylation of the mitogen-activated extracellular signal-regulated activating kinase and its substrate extracellular signal regulated kinase, leading to diminished production of IL-2. Second, IFN-alpha treatment of primary CD4+ T cells reduced proliferative response to stimulation in the presence of exogenous IL-2 by markedly decreasing mRNA synthesis and surface expression of CD25 (alpha-chain), a critical component of the IL-2R complex. These results may be relevant for the antitumor effects of IFN-alpha and may help us to better understand its detrimental role in the inhibition of proliferation of the bulk of CD4+ T cells (uninfected cells) in HIV-infected persons, who are known to overproduce IFN-alpha. PMID- 10679064 TI - Stat6 regulation of in vivo IL-4 responses. AB - Although in vitro development of a Th2 response from naive CD4+ T cells is Stat6 dependent, mice immunized with a goat Ab to mouse IgD have been reported to produce a normal primary IL-4 response in Stat6-deficient mice. Experiments have now been performed with mice immunized with more conventional Ags or inoculated with nematode parasites to account for this apparent discrepancy. The ability of an immunogen to induce a primary in vivo IL-4 response in Stat6-deficient mice was found to vary directly with its ability to induce a strong type 2 cytokine biased response in normal mice. Even immunogens, however, that induce strong primary IL-4 responses in Stat6-deficient mice induce poor memory IL-4 responses in these mice. Consistent with this, Stat6-deficient CD4+ T cells make relatively normal IL-4 responses when stimulated in vitro for 3 days with anti-CD3 and anti CD28, but poor IL-4 responses if they are later restimulated with anti-CD3. Thus, Stat6 signaling enhances primary IL-4 responses that are made as part of a type 0 cytokine response (mixed type 1 and type 2) and is required for normal development or survival of Th2 memory cells. PMID- 10679065 TI - Identification of a membrane Ig-induced p38 mitogen-activated protein kinase module that regulates cAMP response element binding protein phosphorylation and transcriptional activation in CH31 B cell lymphomas. AB - The cAMP response element (CRE) binding protein (CREB) is emerging as a key regulatory factor of gene transcription in B lymphocytes; however, the postreceptor pathways that regulate CREB activity and CRE-dependent gene transcription remain largely undefined. We investigated B cell Ag receptor (BCR) mediated phosphorylation and activation of CREB in the surface IgM+ CH31 B cell lymphoma, which undergoes Ag-dependent cell death. The activity of p38 mitogen activated protein kinase (MAPK) was increased in response to BCR ligation. Phosphorylation of CREB on serine 133, a modification that positively regulates its trans-activation, was concomitantly increased. Inhibition of p38 MAPK by pretreating CH31 B cells with the highly specific bicyclic imidazole inhibitor, SB203580, reduced BCR-induced CREB phosphorylation. BCR cross-linking also led to increased MAPK-activated protein kinase-2 activity, an enzyme that lies immediately downstream from p38 MAPK; MAPK-activated protein kinase-2 immune complexes phosphorylated a peptide substrate containing the CREB serine 133 phosphoacceptor motif. Given the role of CREB in regulating junB gene expression in mature B lymphocytes, we examined whether p38 MAPK activity was necessary for CRE-dependent junB transcription in CH31 B cells. BCR ligation led to increased junB mRNA levels, which were significantly reduced in CH31 B cells pretreated with SB203580. Activation of a CRE-dependent junB promoter/chloramphenicol acetyltransferase (CAT) reporter gene by the BCR was also blocked by SB203580. Similarly, inhibition of p38 MAPK in surface IgM+ WEHI-231 B cell lymphomas resulted in reduced BCR-induced junB mRNA expression and junB promoter activation. The results implicate a p38 MAPK pathway in BCR-mediated CREB phosphorylation and junB transcriptional activation in B cell lymphomas. PMID- 10679066 TI - 4-1BB costimulation is required for protective anti-viral immunity after peptide vaccination. AB - Peptide vaccination induces T cell activation and cytotoxic T cell development. In an effort to understand what factors can improve immune responses to peptide vaccination, the role of 4-1BB (CD137) costimulation was examined, since 4-1BB has been shown to promote T cell responses in other systems. 4-1BBL-deficient (-/ ) and wild-type (+/+) mice were immunized with a lipidated lymphocytic choriomeningitis virus (LCMV) peptide NP396-404. Analysis of peptide-specific responses early after immunization by CTL assay, intracellular IFN-gamma staining, and IFN-gamma enzyme-linked immunospot assay (ELISPOT) indicated that CD8 T cell responses were reduced 3- to 10-fold in the absence of 4-1BB costimulation. Moreover, when agonistic anti-4-1BB Ab was given, CD8 T cell responses in 4-1BBL-/- mice were augmented to levels similar to those in 4 1BBL+/+ mice. Two months after immunization, 4-1BBL+/+ mice still had epitope specific cells and were protected against viral challenge, demonstrating that peptide vaccination can induce long-term protection. In fact, 70% of CD8 T cells were specific for the immunizing peptide after viral challenge, demonstrating that strong, epitope-specific CD8 T cell responses are generated after peptide vaccination. In contrast, peptide-immunized 4-1BBL-/- mice had fewer epitope specific cells and were impaired in their ability to resolve the infection. These results show that immunization with a single LCMV peptide provides long term protection against LCMV infection and point to costimulatory molecules such as 4 1BB as important components for generating protective immunity after vaccination. PMID- 10679067 TI - MEK activity regulates negative selection of immature CD4+CD8+ thymocytes. AB - CD4+CD8+ thymocytes are either positively selected and subsequently mature to CD4 single positive (SP) or CD8 SP T cells, or they die by apoptosis due to neglect or negative selection. This clonal selection is essential for establishing a functional self-restricted T cell repertoire. Intracellular signals through the three known mitogen-activated protein (MAP) kinase pathways have been shown to selectively guide positive or negative selection. Whereas the c-Jun N-terminal kinase and p38 MAP kinase regulate negative selection of thymocytes, the extracellular signal-regulated kinase (ERK) pathway is required for positive selection and T cell lineage commitment. In this paper, we show that the MAP/ERK kinase (MEK)-ERK pathway is also involved in negative selection. Thymocytes from newborn TCR transgenic mice were cultured with TCR/CD3epsilon-specific Abs or TCR specific agonist peptides to induce negative selection. In the presence of the MEK-specific pharmacological inhibitors PD98059 or UO126, cell recovery was enhanced and deletion of DP thymocytes was drastically reduced. Furthermore, development of CD4 SP T cells was blocked, but differentiation of mature CD8 SP T cells proceeded in the presence of agonist peptides when MEK activity was blocked. Thus, our data indicate that the outcome between positively and negatively selecting signals is critically dependent on MEK activity. PMID- 10679068 TI - Qualitative changes accompany memory T cell generation: faster, more effective responses at lower doses of antigen. AB - The generation of memory T cells is critically important for rapid clearance and neutralization of pathogens encountered previously by the immune system. We have studied the kinetics of response and Ag dose requirements for proliferation and cytokine secretion of CD4+ memory T cells to examine whether there are qualitative changes which might lead to improved immunity. TCR Tg CD4+ T cells were primed in vitro and transferred into T cell-deficient hosts. After 6 or more weeks, the persisting T cells were exclusively small resting cells with a memory phenotype: CD44high CD62L+/- CD25-. Memory CD4 T cells showed a similar pattern of response as naive cells to peptide analogues with similar Ag dose requirements for IL-2 secretion. However, memory cells (derived from both Th2 and Th1 effectors) displayed faster kinetics of cytokine secretion, cell division, and proliferation, enhanced proliferation in response to low doses of Ag or peptide analogues, and production of IL-4, IL-5, and IFN-gamma. These results suggest there is a much more efficient response of CD4 memory T cells to Ag re-exposure and that the expanded functional capacity of memory cells will promote a rapid development of effector functions, providing more rapid and effective immunity. PMID- 10679069 TI - Exclusive Th2 primary effector function in spleens but mixed Th1/Th2 function in lymph nodes of murine neonates. AB - Recent studies have shown that neonatal mice are competent to develop mature, Ag specific Th1 function in situ. However, under many conditions, Th2 responses dominate in the neonate, while Th1 responses are more prevalent in adults. To compare further the immune responses of neonates and adults, we used the enzyme linked immunospot method to measure the frequencies of primary Th1/Th2 effectors generated in situ in the spleens and lymph nodes. As assessed by the detection of IFN-gamma- or IL-4-producing cells, adults developed mixed Th1/Th2 responses in both organs. Neonatal lymph nodes contained mature frequencies of IFN-gamma- and IL-4-producing cells. In striking contrast, while mature frequencies of Th2 cells developed in neonatal spleens, virtually no IFN-gamma-secreting cells were detected. Exclusive Th2 function was observed in both BALB/c and C57BL/6 neonates, strains in which the Th2 and Th1 lineages, respectively, are favored in adults. Although Th1 effectors were virtually undetectable, the addition of rIL 12 boosted the frequency of IFN-gamma-secreting cells to adult levels. Therefore, Th1 effectors apparently developed in situ, but Th1 effector function either was not promoted or was inhibited upon subsequent exposure to the Ag in culture. Together, these results indicate that the quality of a primary Th response in neonates is strongly dependent on the site of initial Ag exposure; responses initiated in the lymph nodes are mixed Th1/Th2, whereas responses occurring in the spleen are heavily Th2 biased. PMID- 10679070 TI - The density of peptides displayed by dendritic cells affects immune responses to human tyrosinase and gp100 in HLA-A2 transgenic mice. AB - Several HLA-A*0201-restricted peptide epitopes that can be used as targets for active immunotherapy have been identified within melanocyte differentiation proteins. However, uncertainty exists as to the most effective way to elicit CD8+ T cells with these epitopes in vivo. We report the use of transgenic mice expressing a derivative of HLA-A*0201, and dendritic cells, to enhance the activation of CD8+ T cells that recognize peptide epitopes derived from human tyrosinase and glycoprotein 100. We find that by altering the cell surface density of the immunizing peptide on the dendritic cells, either by pulsing with higher concentrations of peptide, or by changing the MHC-peptide-binding affinity by generating variants of the parent peptides, the size of the activated CD8+ T cell populations can be modulated in vivo. Significantly, the density of peptide that produced the largest response was less than the maximum density achievable through short-term peptide pulsing. We have also found, however, that while some variant peptides are effective at eliciting both primary and recall CD8+ T cell responses that can recognize the parental epitope, other variant epitopes lead to the outgrowth of CD8+ T cells that only recognize the variant. HLA-A*0201 transgenic mice provide an important model to define which peptide variants are most likely to stimulate CD8+ T cell populations that recognize the parental, melanoma-specific peptide. PMID- 10679071 TI - Heat shock protein 70 is able to prevent heat shock-induced resistance of target cells to CTL. AB - Heat shock or transfection with heat shock protein 70 (Hsp70) genes has been shown to protect tumor cell lines against immune mechanisms of cytotoxicity. We have reported previously that heat shock confers resistance to CTL in the rat myeloma cell line Y3 that is Hsp70 defective. Evidence is now presented that Hsp70 is able to prevent the induction of the resistant phenotype. In Con A stimulated lymphocytes and in lymphocyte x Y3 somatic cell hybrid clones a severe, non-Hsp70-inducing heat shock elicits resistance to CTL in contrast to a heat shock that results in Hsp70 expression. Thus, Hsp70 expression appears to be negatively associated with the development of resistance. Furthermore, loading of Y3 cells with recombinant Hsp70 protein before heat shock is able to prevent resistance. Because apoptosis induced in Y3 cells by heat shock is not affected, Hsp70 appears to interfere selectively with the CTL-induced lethal pathway that is found to be calcium but not caspase dependent. It is suggested that after heat shock Hsp70 enhances the CTL-induced apoptotic pathway by chaperoning certain proteins in the target cell that are involved in the execution of cell death. Thus, although shown to confer protection against many cytotoxic mechanisms, Hsp70 does not appear to be generally cytoprotective. This observation could also be of relevance when interpreting the effectiveness of tumor immunity. PMID- 10679072 TI - CpG-DNA activates in vivo T cell epitope presenting dendritic cells to trigger protective antiviral cytotoxic T cell responses. AB - MHC class I-restricted T cell epitopes lack immunogenicity unless aided by IFA or CFA. In an attempt to circumvent the known inflammatory side effects of IFA and CFA, we analyzed the ability of immunostimulatory CpG-DNA to act as an adjuvant for MHC class I-restricted peptide epitopes. Using the immunodominant CD8 T cell epitopes, SIINFEKL from OVA or KAVYNFATM (gp33) from lymphocytic choriomeningitis virus glycoprotein, we observed that CpG-DNA conveyed immunogenicity to these epitopes leading to primary induction of peptide-specific CTL. Furthermore, vaccination with the lymphocytic choriomeningitis virus gp33 peptide triggered not only CTL but also protective antiviral defense. We also showed that MHC class I-restricted peptides are constitutively presented by immature dendritic cells (DC) within the draining lymph nodes but failed to induce CTL responses. The use of CpG-DNA as an adjuvant, however, initiated peptide presenting immature DC progression to professional licensed APC. Activated DC induced cytolytic CD8 T cells in wild-type mice and also mice deficient of Th cells or CD40 ligand. CpG DNA thus incites CTL responses toward MHC class I-restricted T cell epitopes in a Th cell-independent manner. Overall, these results provide new insights into CpG DNA-mediated adjuvanticity and may influence future vaccination strategies for infectious and perhaps tumor diseases. PMID- 10679073 TI - HLA-DR-mediated apoptosis susceptibility discriminates differentiation stages of dendritic/monocytic APC. AB - Professional APC are characterized by their ability to present peptide via HLA class II in the presence of costimulatory molecules (CD40, CD80, and CD86). The efficiency of Ag presentation can be classed as follows: mature dendritic cells (DC) are most efficient, immature DC and macrophages are intermediate, and monocytes are considered poor APC. There is a large body of evidence demonstrating that HLA-DR transmits signals in the APC. In this study, we have addressed the question of the outcome of HLA-DR signals on APC of the monocyte/DC lineages throughout their differentiation from immature to mature APC. DC were generated from both monocytes and CD34+ cells of the same individual, macrophages were differentiated from monocytes. Immunophenotypical analysis clearly distinguished these populations. HLA-DR-mediated signals led to marked apoptosis in mature DC of either CD34 or monocytic origin. Significantly less apoptosis was observed in immature DC of either origin. Nonetheless, even immature DC were more susceptible to HLA-DR-mediated apoptosis than macrophages, whereas monocytes were resistant to HLA-DR-mediated apoptosis. The mechanism of HLA-DR-mediated apoptosis was independent of caspase activation. Taken together, these data lead to the notion that signals generated via HLA-DR lead to the demise of mature professional APC, thereby providing a means of limiting the immune response. PMID- 10679074 TI - Sensitivity difference to the suppressive effect of prostaglandin E2 among mouse strains: a possible mechanism to polarize Th2 type response in BALB/c mice. AB - PGE2 has been shown to play a prominent role in regulating Th1 and Th2 type responses. We studied the role of PGE2 in IFN-gamma production by Staphylococcus aureus Cowan I-stimulated spleen cells from several mouse strains such as BALB/c, C3H/HeN, and C57BL/6. When spleen cells were pretreated with indomethacin (cyclooxygenase (COX)-1 and COX-2 inhibitor) or NS-398 (COX-2-specific inhibitor), S. aureus Cowan I -induced IFN-gamma production was increased more markedly in spleen cells from BALB/c mice than from C3H/HeN and C57BL/6 mouse. However, PGE2 production was not significantly different among spleen cells from three mouse strains. When various concentrations of PGE2 were exogeneously added to spleen cells, PGE2 showed a stronger suppressive effect on IFN-gamma production in spleen cells from BALB/c mice than from other strains of mice. This suppressive effect of PGE2 in BALB/c mice mainly depended on IL-12p70 production by APCs. More PGE2 binding sites were found in BALB/c spleen cells than in C3H/HeN spleen cells, indicating that the sensitivity difference to the suppressive effect of PGE2 was due to the difference of the number of PGE2 receptors. The administration of NS-398 into BALB/c mice enhanced Ag-specific IFN gamma production, but not IL-4 production. This effect is the same as IL-12 administration in vivo. From these results, we propose that the modulation of PGE2 is important for Th1 activation via IFN-gamma and IL-12p70 production in vitro and in vivo and that PGE2 is one of the pivotal factors in the Th2-dominant immune response in BALB/c mice. PMID- 10679075 TI - RANTES binding and down-regulation by a novel human herpesvirus-6 beta chemokine receptor. AB - The human herpesvirus 6 (HHV-6) U51 gene defines a new family of betaherpesvirus specific genes encoding multiple transmembrane glycoproteins with similarity to G protein-coupled receptors, in particular, human chemokine receptors. These are distinct from the HHV-6 U12 and HCMV US28 family. In vitro transcription and translation as well as transient cellular expression of U51 showed properties of a multiple transmembrane protein with a 30-kDa monomer as well as high m.w. aggregates or oligomers. Transient cellularly expressed U51 also appeared to form dimeric intermediates. Despite having only limited sequence similarity to chemokine receptors, U51 stably expressed in cell lines showed specific binding of the CC chemokine RANTES and competitive binding with other beta chemokines, such as eotaxin; monocyte chemoattractant protein 1, 3, and 4; as well as the HHV 8 chemokine vMIPII. In epithelial cells already secreting RANTES, U51 expression resulted in specific transcriptional down-regulation. This correlated with reduced secretion of RANTES protein into the culture supernatants. Regulation of RANTES levels may alter selective recruitment of circulating inflammatory cells that the virus can infect and thus could mediate the systemic spread of the virus from initial sites of infection in epithelia. Alternatively, chemokine regulation could modulate a protective inflammatory response to aid the spread of virus by immune evasion. Such mimicry, by viral proteins, of host receptors leading to down-regulation of chemokine expression is a novel immunomodulatory mechanism. PMID- 10679076 TI - 1 Alpha,25-dihydroxyvitamin D3 inhibits differentiation, maturation, activation, and survival of dendritic cells leading to impaired alloreactive T cell activation. AB - 1 Alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3), the active form of vitamin D3, is a potent immunomodulatory agent. Here we show that dendritic cells (DCs) are major targets of 1,25(OH)2D3-induced immunosuppressive activity. 1,25(OH)2D3 prevents the differentiation in immature DCs of human monocytes cultured with GM-CSF and IL-4. Addition of 1,25(OH)2D3 during LPS-induced maturation maintains the immature DC phenotype characterized by high mannose receptor and low CD83 expression and markedly inhibits up-regulation of the costimulatory molecules CD40, CD80, and CD86 and of class II MHC molecules. This is associated with a reduced capacity of DCs to activate alloreactive T cells, as determined by decreased proliferation and IFN-gamma secretion in mixed leukocyte cultures. 1, 25(OH)2D3 also affects maturing DCs, leading to inhibition of IL-12p75 and enhanced IL-10 secretion upon activation by CD40 ligation. In addition, 1,25(OH)2D3 promotes the spontaneous apoptosis of mature DCs. The modulation of phenotype and function of DCs matured in the presence of 1,25(OH)2D3 induces cocultured alloreactive CD4+ cells to secrete less IFN-gamma upon restimulation, up-regulate CD152, and down-regulate CD154 molecules. The inhibition of DC differentiation and maturation as well as modulation of their activation and survival leading to T cell hyporesponsiveness may explain the immunosuppressive activity of 1, 25(OH)2D3. PMID- 10679077 TI - NK1.1+ T cells in the liver arise in the thymus and are selected by interactions with class I molecules on CD4+CD8+ cells. AB - NK1.1+ T cells represent a specialized T cell subset specific for CD1d, a nonclassical MHC class I-restricting element. They are believed to function as regulatory T cells. NK1.1+ T cell development depends on interactions with CD1d molecules presented by hematopoietic cells rather than thymic epithelial cells. NK1.1+ T cells are found in the thymus as well as in peripheral organs such as the liver, spleen, and bone marrow. The site of development of peripheral NK1.1+ T cells is controversial, as is the nature of the CD1d-expressing cell that selects them. With the use of nude mice, thymectomized mice reconstituted with fetal liver cells, and thymus-grafted mice, we provide direct evidence that NK1.1+ T cells in the liver are thymus dependent and can arise in the thymus from fetal liver precursor cells. We show that the class I+ (CD1d+) cell type necessary to select NK1.1+ T cells can originate from TCRalpha-/- precursors but not from TCRbeta-/- precursors, indicating that the selecting cell is a CD4+CD8+ thymocyte. 5-Bromo-2'-deoxyuridine-labeling experiments suggest that the thymic NK1.1+ T cell population arises from proliferating precursor cells, but is a mostly sessile population that turns over very slowly. Since liver NK1.1+ T cells incorporate 5-bromo-2'-deoxyuridine more rapidly than thymic NK1.1+ T cells, it appears that liver NK1.1+ T cells either represent a subset of thymic NK1.1+ T cells or are induced to proliferate after having left the thymus. The results indicate that NK1.1+ T cells, like conventional T cells, arise in the thymus where they are selected by interactions with restricting molecules. PMID- 10679078 TI - Role of IL-6 in directing the initial immune response to schistosome eggs. AB - The eggs of Schistosoma mansoni are strong inducers of a Th2 response, and previous work has shown that Ag-specific IL-6 is produced within 24 h after the injection of eggs into mice. Investigations to determine the role of IL-6 in orchestrating the early response to schistosome eggs have revealed that IL-12 is rapidly produced in lymph node cell cultures from egg-injected mice. This "early" IL-12 primes for the production of IL-6 and IFN-gamma, for in IL-12-/- mice egg injection fails to stimulate increased production of either of these cytokines. Furthermore, IL-6 also up-regulates IL-10 production which, together with IL-6, negatively regulates IL-12 and IFN-gamma production. Finally, IL-10 down regulates the production of its inducer, IL-6. These data indicate that the anti inflammatory role of IL-6 may be effected through negative regulation of type 1 (IFN-gamma) and type 1-associated (IL-12) cytokines either directly (by IL-6) or indirectly (through the induction of IL-10) and suggest that one mechanism by which eggs may support the development of Th2 responses is through the negative regulation of the type 1 response. PMID- 10679079 TI - Specific B cell tolerance is induced by cyclosporin A plus donor-specific blood transfusion pretreatment: prolonged survival of MHC class I disparate cardiac allografts. AB - Donor-specific blood transfusion (DST), designed to prolong allograft survival, sensitized recipients of the high-responder PVG-RT1u strain, resulting in accelerated rejection of MHC-class I mismatched (PVG-R8) allografts. Rejection was found to be mediated by anti-MHC class I (Aa) alloantibody. By pretreating recipients 4 wk before grafting with cyclosporin A (CsA) daily (x7), combined with once weekly (x4) DST, rejection was prevented. The investigation explores the mechanism for this induced unresponsiveness. CD4 T cells purified from the thoracic duct of CsA/DST-pretreated RT1u rats induced rejection when transferred to R8 heart-grafted RT1u athymic nude recipients, indicating that CD4 T cells were not tolerized by the pretreatment. To determine whether B cells were affected, nude recipients were pretreated, in the absence of T cells, with CsA/DST (or CsA/third party blood) 4 wk before grafting. The subsequent transfer of normal CD4 T cells induced acute rejection of R8 cardiac allografts in third party- but not DST-pretreated recipients; prolonged allograft survival was reversed by the cotransfer of B cells with the CD4 T cells. Graft survival correlated with reduced production of anti-MHC class I (Aa) cytotoxic alloantibody. The results indicated that the combined pretransplant treatment of CsA and DST induced tolerance in allospecific B cells independently of T cells. The resulting suppression of allospecific cytotoxic Ab correlated with the survival of MHC class I mismatched allografts. The induction of B cell tolerance by CsA has important implications for clinical transplantation. PMID- 10679080 TI - Engrafting costimulator molecules onto tumor cell surfaces with chelator lipids: a potentially convenient approach in cancer vaccine development. AB - The genetic modification of cells to develop cell-based vaccines and to modulate immune responses in vivo can be risky and inconvenient to perform in clinical situations. A novel chelator lipid, nitrilotriacetic acid di-tetradecylamine (NTA DTDA) that, via the NTA group has high affinity for 6His peptide, was used to directly anchor recombinant forms of T cell costimulatory molecules containing a C-terminal 6-His sequence onto tumor cell surfaces. Initial experiments using murine P815 tumor cells established the optimum conditions for incorporating NTA DTDA onto the membranes of cells. P815 cells with incorporated NTA-DTDAbound hexahistidine-(6His)-tagged forms of the extracellular domains of murine B7.1 and CD40 (B7.1-6H and CD40-6H) at very high levels (fluorescence 200-300-fold above background), and both proteins could be anchored onto the cells simultaneously. Significant loss of the anchored or "engrafted" protein occurred through membrane internalization following culture of the cells under physiological conditions, but P815 cells with engrafted B7.1-6H and/or CD40-6H stimulated the proliferation of allogenic and syngeneic splenic T cells in vitro, and generated cytotoxic T cells when used as vaccines in syngeneic animals. Furthermore, the immunization of syngeneic mice with P815 cells engrafted with B7.1-6H or with B7. 1-6H and CD40-6H induced protection against challenge with the native P815 tumor. The results indicate that the use of chelator lipids like NTD-DTDA to engraft costimulatory and/or other molecules onto cell membranes could provide a convenient alternative to transfection in the development of cell-based vaccines and for modulation of immune function. PMID- 10679081 TI - Low CD86 expression in the nonobese diabetic mouse results in the impairment of both T cell activation and CTLA-4 up-regulation. AB - The nonobese diabetic (NOD) mouse spontaneously develops autoimmune insulin dependent diabetes mellitus and serves as a model for human type I diabetes. NOD spleen cells proliferate to a lesser extent than those from C57BL/6 and BALB/c mice in response to anti-CD3. To investigate the cause of this reduced T cell proliferation, costimulatory molecule expression was investigated. It was found that NOD macrophages, dendritic cells, and T cells, but not B cells, expressed lower basal levels of CD86, but not CD80, CD28, or CD40, compared with C57BL/6 and BALB/c. This low CD86 expression was not dependent on the MHC haplotype or on diabetes development since the NOD-related, diabetes-free mouse strains NON (H 2nb1) and NOR (H-2g7) exhibited similar low levels of CD86 expression and proliferation. Furthermore, following activation, the relative up-regulation of CTLA-4, as compared with CD28, was more pronounced on C57BL/6 and BALB/c T cells as shown by an increased CTLA-4/CD28 ratio. This activation-induced increase in the CTLA-4/CD28 ratio was markedly reduced on NOD T cells compared with the other two strains. The low CD86 expression in NOD mice may account for the reduced increase in both proliferation and the CTLA-4/CD28 ratio, since reducing CD86 expression in C57BL/6 and BALB/c cultures to NOD levels significantly reduces the proliferation and the CTLA-4/CD28 ratio. Therefore, we propose that a low level of CD86 expression in the NOD mouse contributes to a defective regulation of autoreactive T cells by preventing the full activation of T cells and therefore the up-regulation of CTLA-4. PMID- 10679082 TI - IL-10 contributes to the inhibition of contact hypersensitivity in mice treated with photodynamic therapy. AB - We have explored the effect of photodynamic therapy (PDT) with verteporfin on the induction and expression of contact hypersensitivity (CHS) to 2,4 dinitrofluorobenzene (DNFB) in normal mice and IL-10-deficient mice. Our results indicate that DNFB sensitized mice given PDT with verteporfin and whole body red light irradiation exhibited a significant reduction in CHS compared with control animals. Administration of rIL-12 reversed the effect(s) of PDT as did treatment of mice with anti-IL-10-neutralizing Ab. Knockout mice deficient in IL-10 were found to be resistant to the inhibitory effects of PDT. In vitro proliferative responses using spleen cells from DNFB-sensitized and PDT-treated mice showed a significantly lower response to DNBS as compared with cells from DNFB-sensitized mice or DNFB and PDT-treated IL-10-deficient mice. Finally, naive mice exposed to PDT exhibited an increase in skin IL-10 levels, which peaked between 72 and 120 h post-PDT. Together these data support the role of IL-10 as a key modulator in the inhibition of the CHS response by whole body PDT. PMID- 10679083 TI - Stage-specific expression of mucosal addressin cell adhesion molecule-1 during embryogenesis in rats. AB - Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is essential for lymphocyte trafficking to gut-associated lymphoid tissues and is implicated in inflammatory disorders in the gut and pancreatic islets. In this study, we examined the functional role of MAdCAM-1 during rat ontogeny using newly generated specific mAb. As previously observed in mice and humans, MAdCAM-1 was preferentially expressed in high endothelial venules (HEV) in gut-associated lymphoid tissues and venules of lamina propria in adult rats. Lymphocyte rolling and adhesion on HEV in Peyer's patches (PP) were completely abrogated with neutralizing anti MAdCAM-1 mAb, in agreement with the notion that MAdCAM-1 is the principal HEV ligand for lymphocyte rolling and adhesion in adult PP. In the developing gastrointestinal tract, MAdCAM-1 was widely expressed in the venules of the lamina propria of fetal rats. In addition, MAdCAM-1 was also expressed in follicular dendritic cells in the neonatal PP. Interestingly, MAdCAM-1 expression was found also in nonmucosal tissues during ontogeny. MAdCAM-1 was transiently expressed in blood vascular endothelial cells in the fetal skin and neonatal thymus. Notably, MAdCAM-1-positive blood vessels were localized mainly in the cortico-medullary junction in the neonatal thymus and about 10-20% of thymocytes, most of which were either CD4, CD8 double positive or single positive specifically reacted with soluble MAdCAM-1 via integrin alpha4beta7. After birth, MAdCAM-1 expression in thymus blood vessels disappeared and concomitantly, the soluble MAdCAM-1-reactive thymocytes were rapidly down-regulated. Our results suggest that MAdCAM-1 functions as a vascular addressin in not only mucosal, but also nonmucosal lymphoid tissues during ontogeny. PMID- 10679084 TI - The accumulation of dendritic cells in the lung is impaired in CD18-/- but not in ICAM-1-/- mutant mice. AB - Bone marrow-derived dendritic cell (DC) precursors migrate via the blood stream to peripheral tissues to adopt their sentinel function. To identify factors facilitating their emigration to the lung, mutant mice deficient in E-selectin, P selectin, E/P-selectin, ICAM-1, or CD18 and their respective controls were examined. DCs and monocytes/macrophages were immunolabeled with M5/114 and MOMA-2 mAbs, respectively, and quantified morphometrically. Of these genotypes, the numbers of DC and MOMA-2+ cells were significantly less only in the lungs of CD18 /- mice by 68 and 35% in alveolar walls and by 28 and 26% in venous walls, respectively. DCs were reduced by 30 and 41% around large and small airways, respectively, but the number of MOMA-2+ cells in these locations was not significantly different from controls. Ablation of a single gene may be associated with augmented expression of other, related gene products. Therefore, we examined the expression of VCAM-1. Increased numbers of arteries exhibited continuous luminal VCAM-1 staining in both CD18-/- and ICAM-1-/- mutants. VCAM-1 expression was absent in pulmonary capillaries and unchanged in veins. These data suggest that under nonperturbing conditions, CD18-mediated adhesion is required for the full complement of DC precursors to accumulate in the lungs. However, the defect in CD18-/- mice is partial, suggesting that CD18-independent adhesion occurs. The alternative pathway may involve VLA-4/VCAM-1 in arteries and venules but not in capillaries. The smaller defect in ICAM-1-/- mice suggests that the CD11/CD18 complex recognizes ligands other than ICAM-1 at some sites. PMID- 10679085 TI - Specific inhibition of glucocorticoid-induced thymocyte apoptosis by substance P. AB - Glucocorticoids (GC) are strong inducers of thymocyte apoptosis. In the present study we looked into the possibility that the neuropeptide substance P (SP) might serve as an antagonist to GC-induced apoptosis. Indeed, SP inhibited hydrocortisone (HC)-induced apoptosis of CD4+CD8+ thymocytes in mice, both in vivo and in vitro. It also inhibited HC-induced apoptosis in the T cell hybridoma line 2B4.11, which is sensitive to GC. The inhibitory effect was complete if SP was given with HC or within 1 h after it; partial inhibitory effect could be seen at 2 h and no effect at 3 h. The presence of the SP antagonist nullified SP effect. The effect was specific to both components of the system (i.e., HC as apoptosis inducer and SP as its inhibitor), as judged from comparison to three other apoptosis-inducing means (irradiation, thymic epithelial cells, or retinoic acid), and to two other neuropeptides (somatostatin and vasoactive intestinal peptide). SP/HC antagonism was further demonstrated in two relevant molecular events: 1) HC augmented GC receptor production in our cell system and this was inhibited by SP; and 2) HC reduced the expression of the transcription factor NF kappaB, SP increased it and when both were present, SP effect dominated. On the other hand, the level of IkappaB (NF-kappaB inhibitory molecule) was decreased by SP, preserved at a relatively high level with HC, and when both SP and HC were present, SP effect dominated. The intensity of SP effect, both in vivo and in vitro, its specificity, its inhibition by SP antagonist, as well as the previously documented presence of SP and its receptor in the thymus suggest that SP might be a physiological antagonist of the potent thymocyte apoptosis induced by GC. PMID- 10679086 TI - Maturation and trafficking of monocyte-derived dendritic cells in monkeys: implications for dendritic cell-based vaccines. AB - Human dendritic cells (DC) have polarized responses to chemokines as a function of maturation state, but the effect of maturation on DC trafficking in vivo is not known. We have addressed this question in a highly relevant rhesus macaque model. We demonstrate that immature and CD40 ligand-matured monocyte-derived DC have characteristic phenotypic and functional differences in vitro. In particular, immature DC express CC chemokine receptor 5 (CCR5) and migrate in response to macrophage inflammatory protein-1alpha (MIP-1alpha), whereas mature DC switch expression to CCR7 and respond exclusively to MIP-3beta and 6Ckine. Mature DC transduced to express a marker gene localized to lymph nodes after intradermal injection, constituting 1.5% of lymph node DC. In contrast, cutaneous DC transfected in situ via gene gun were detected in the draining lymph node at a 20-fold lower frequency. Unexpectedly, the state of maturation at the time of injection had no influence on the proportion of DC that localized to draining lymph nodes, as labeled immature and mature DC were detected in equal numbers. Immature DC that trafficked to lymph nodes underwent a significant up-regulation of CD86 expression indicative of spontaneous maturation. Moreover, immature DC exited completely from the dermis within 36 h of injection, whereas mature DC persisted in large numbers associated with a marked inflammatory infiltrate. We conclude that in vitro maturation is not a requirement for effective migration of DC in vivo and suggest that administration of Ag-loaded immature DC that undergo natural maturation following injection may be preferred for DC-based immunotherapy. PMID- 10679088 TI - Lymphotoxin-alpha-dependent spleen microenvironment supports the generation of memory B cells and is required for their subsequent antigen-induced activation. AB - Lymphotoxin alpha-deficient (LTalpha-/-) mice show dramatically reduced IgG responses after either primary or secondary immunizations with sheep red blood cells (SRBC). When splenocytes from SRBC-primed wild-type donor mice were infused into irradiated naive wild-type recipient mice, they generated a robust memory IgG response, but not when infused into LTalpha-/- recipients, indicating that the microenvironment that develops in LTalpha-/- mice is incompetent to support the activation of this memory response. When irradiated wild-type mice were reconstituted with splenocytes from primed LTalpha-/- donors and then challenged with the same immunizing Ag, no memory response was observed, indicating further that memory cells could not be generated in the LTalpha-/- environment. To address which lymphocyte subsets were impaired in the LTalpha-/- mice, we performed reconstitution experiments using a hapten/carrier system and T cells and B cells from different primed donors. There was no detectable defect in either the generation or expression of memory T cells from LTalpha-/- donors. In contrast, B cells were not primed for memory in the microenvironment of LTalpha-/ mice. Additionally, primed wild-type memory B cells could not express a memory IgG response in the LTalpha-/- microenvironment. Thus, splenic white pulp structure, which depends on the expression of LTalpha for its development and maintenance, is needed to support the generation of memory B cells and to permit existing memory B cells to express an isotype switched memory Ig response following antigenic challenge. PMID- 10679087 TI - NOD/LtSz-Rag1null mice: an immunodeficient and radioresistant model for engraftment of human hematolymphoid cells, HIV infection, and adoptive transfer of NOD mouse diabetogenic T cells. AB - Development of a small animal model for the in vivo study of human immunity and infectious disease remains an important goal, particularly for investigations of HIV vaccine development. NOD/Lt mice homozygous for the severe combined immunodeficiency (Prkdcscid) mutation readily support engraftment with high levels of human hematolymphoid cells. However, NOD/LtSz-scid mice are highly radiosensitive, have short life spans, and a small number develop functional lymphocytes with age. To overcome these limitations, we have backcrossed the null allele of the recombination-activating gene (Rag1) for 10 generations onto the NOD/LtSz strain background. Mice deficient in RAG1 activity are unable to initiate V(D)J recombination in Ig and TCR genes and lack functional T and B lymphocytes. NOD/LtSz-Rag1null mice have an increased mean life span compared with NOD/LtSz-scid mice due to a later onset of lymphoma development, are radioresistant, and lack serum Ig throughout life. NOD/LtSz-Rag1null mice were devoid of mature T or B cells. Cytotoxic assays demonstrated low NK cell activity. NOD/LtSz-Rag1null mice supported high levels of engraftment with human lymphoid cells and human hemopoietic stem cells. The engrafted human T cells were readily infected with HIV. Finally, NOD/LtSz-Rag1null recipients of adoptively transferred spleen cells from diabetic NOD/Lt+/+ mice rapidly developed diabetes. These data demonstrate the advantages of NOD/LtSz-Rag1null mice as a radiation and lymphoma-resistant model for long-term analyses of engrafted human hematolymphoid cells or diabetogenic NOD lymphoid cells. PMID- 10679089 TI - Oral administration of hapten inhibits in vivo induction of specific cytotoxic CD8+ T cells mediating tissue inflammation: a role for regulatory CD4+ T cells. AB - We investigated whether oral tolerance could block the development of an inflammatory response mediated by CD8+ T cells, using a mouse model of oral tolerance of contact sensitivity (CS) to the hapten 2, 4-dinitrofluorobenzene (DNFB). In this system, the skin inflammatory response is initiated by hapten specific class I-restricted cytotoxic CD8+ T (CTL) cells, independently of CD4 help. Oral delivery of DNFB before skin sensitization blocked the CS response by impairing the development of DNFB-specific CD8+ effector T cells in secondary lymphoid organs. This was shown by complete inhibition of DNFB-specific CTL and proliferative responses of CD8+ T cells, lack of specific IFN-gamma-producing CD8+ T cells, and inability of CD8+ T cells to transfer CS in RAG20/0 mice. RT PCR and immunohistochemical analysis confirmed that recruitment of CD8+ effectors of CS in the skin at the site of hapten challenge was impaired in orally tolerized mice. Sequential anti-CD4 Ab treatment showed that only depletion of CD4+ T cells during the afferent phase of CS abrogated oral tolerance induction by restoring high numbers of specific CD8+ effectors in lymphoid organs, whereas CD4 depletion during the efferent phase of CS did not affect oral tolerance. These data demonstrate that a single intragastric administration of hapten can block in vivo induction of DNFB-specific CD8+ CTL responsible for tissue inflammation and that a subset of regulatory CD4+ T cells mediate oral tolerance by inhibiting expansion of specific CD8+ effectors in lymph nodes. PMID- 10679090 TI - Significant role for Fas in the pathogenesis of autoimmune diabetes. AB - Programmed cell death represents an important pathogenic mechanism in various autoimmune diseases. Type I diabetes mellitus (IDDM) is a T cell-dependent autoimmune disease resulting in selective destruction of the beta cells of the islets of Langerhans. beta cell apoptosis has been associated with IDDM onset in both animal models and newly diagnosed diabetic patients. Several apoptotic pathways have been implicated in beta cell destruction, including Fas, perforin, and TNF-alpha. Evidence for Fas-mediated lysis of beta cells in the pathogenesis of IDDM in nonobese diabetic (NOD) mice includes: 1) Fas-deficient NOD mice bearing the lpr mutation (NOD-lpr/lpr) fail to develop IDDM; 2) transgenic expression of Fas ligand (FasL) on beta cells in NOD mice may result in accelerated IDDM; and 3) irradiated NOD-lpr/lpr mice are resistant to adoptive transfer of diabetes by cells from NOD mice. However, the interpretation of these results is complicated by the abnormal immune phenotype of NOD-lpr/lpr mice. Here we present novel evidence for the role of Fas/FasL interactions in the progression of NOD diabetes using two newly derived mouse strains. We show that NOD mice heterozygous for the FasL mutation gld, which have reduced functional FasL expression on T cells but no lymphadenopathy, fail to develop IDDM. Further, we show that NOD-lpr/lpr mice bearing the scid mutation (NOD-lpr/lpr-scid/scid), which eliminates the enhanced FasL-mediated lytic activity induced by Fas deficiency, still have delayed onset and reduced incidence of IDDM after adoptive transfer of diabetogenic NOD spleen cells. These results provide evidence that Fas/FasL-mediated programmed cell death plays a significant role in the pathogenesis of autoimmune diabetes. PMID- 10679091 TI - The IL-2 receptor promotes lymphocyte proliferation and induction of the c-myc, bcl-2, and bcl-x genes through the trans-activation domain of Stat5. AB - Studies assessing the role of Stat5 in the IL-2 proliferative signal have produced contradictory, and thus inconclusive, results. One factor confounding many of these studies is the ability of IL-2R to deliver redundant mitogenic signals from different cytoplasmic tyrosines on the IL-2R beta-chain (IL-2Rbeta). Therefore, to assess the role of Stat5 in mitogenic signaling independent of any redundant signals, all cytoplasmic tyrosines were deleted from IL-2Rbeta except for Tyr510, the most potent Stat5-activating site. This deletion mutant retained the ability to induce Stat5 activation and proliferation in the T cell line CTLL 2 and the pro-B cell line BA/F3. A set of point mutations at or near Tyr510 that variably compromised Stat5 activation also compromised the proliferative signal and revealed a quantitative correlation between the magnitude of Stat5 activation and proliferation. Proliferative signaling by a receptor mutant with a weak Stat5 activating site could be rescued by overexpression of wt Stat5a or b. Additionally, the ability of this receptor mutant to induce c-myc, bcl-x, and bcl 2 was enhanced by overexpression of wt Stat5. By contrast, overexpression of a version of Stat5a lacking the C-terminal trans-activation domain inhibited the induction of these genes and cell proliferation. Thus, Stat5 is a critical component of the proliferative signal from Tyr510 of the IL-2R and regulates expression of both mitogenic and survival genes through its trans-activation domain. PMID- 10679092 TI - Molecular analysis of the autoantibody response in peptide-induced autoimmunity. AB - Immunization of nonautoimmune BALB/c mice with multimeric DWEYSVWLSN, a peptide mimotope of DNA, induces anti-DNA and other lupus-associated Abs. To further investigate the pathogenesis of the autoantibody response induced by peptide immunization, we generated hybridomas from peptide-immunized mice that bound peptide, dsDNA, cardiolipin, Sm/ribonucleoprotein (RNP), or some combination of these Ags. Analysis of 24 IgM Abs led to the identification of three groups of Abs: 1) Abs reactive with peptide alone, 2) anti-peptide Abs cross-reactive with one or more autoantigens, and 3) autoantibodies that do not bind to peptide. The gene families and particular VH-VL combinations used in those hybridomas binding DNA were similar to those used in the anti-DNA response in spontaneous murine lupus. Another similarity to the spontaneous anti-DNA response was the generation of arginines in the complementarity-determining region-3 of DNA-binding hybridomas. Interestingly, one Ab had the VH-VL combination present in the original R4A anti-DNA Ab used to select the DWEYSVWLSN peptide from a phage display library. Many of the heavy and light chains displayed evidence of somatic mutation, suggesting that they were made by Ag-activated B cells. Analysis of the Ab repertoire in peptide-induced autoimmunity may provide insights into the generation of anti-DNA Abs following exposure to foreign Ag. Furthermore, the recovery of an Ab with the heavy and light chain combination of the Ab originally used to isolate the immunizing peptide confirms the utility of phage display peptide libraries in generating true molecular mimics. PMID- 10679093 TI - An upstream Oct-1- and Oct-2-binding silencer governs B29 (Ig beta) gene expression. AB - The B cell-specific B29 (Igbeta) gene is activated in the earliest B cell precursors and is expressed throughout B cell development. Tissue-specific expression of the murine B29 gene is controlled by a B cell-specific promoter whose activity is governed by a cassette of upstream transcriptional silencers. This study describes a potent new silencer that is located 5' of the previously identified B29 silencer elements, FROG and TOAD. Like these known elements, the new B29 silencer is not restricted to the B29 promoter. Nuclear proteins from all cell lines tested interacted with this A+T-rich sequence, which closely resembled a noncanonical octamer binding motif and also conformed to the consensus sequence for nuclear matrix attachment regions. Interaction of Oct-1 and Oct-2 with the B29 A+T-rich sequence was confirmed using octamer-specific Abs. Oct-1/Oct-2 binding was required for the inhibitory activity of this sequence because mutations that blocked Oct-1/Oct-2 binding also eliminated inhibition of the B29 promoter. This B29 A+T-rich sequence specifically interacted with isolated nuclear matrix proteins in vitro, suggesting that it may also function as a matrix attachment region element. Maintenance of the level of B29 gene expression through the interaction of the minimal promoter and the upstream silencer elements FROG, TOAD, and the A+T-rich Oct-1/Oct-2 binding motif may be essential for normal B cell development and/or function. PMID- 10679094 TI - Regulation of the calcium/NF-AT T cell activation pathway by the D2 domain of CD45. AB - CD45 contains tandem repeated protein tyrosine phosphatase (PTP) domains and is essential for the initiation of the earliest activation events resulting from Ag ligation of the TCR. The second PTP domain (D2) contains four CK2 phosphorylation sites in a unique 19-aa insert, which are targets of CK2 phosphorylation. This study was designed to evaluate the roles of these Ser residues in T cell activation. Transient transfection of the CD45- T cell line, J45.01, with CD45 cDNA incorporating four Ser to Ala (S/A) mutations in the 19-aa insert did not affect the magnitude of NF-AT activation resulting from TCR ligation. However, the basal level of NF-AT activity in unstimulated cells expressing the CD45 S/A mutation was elevated 9- to 10-fold. Increased basal NF-AT was dependent on extracellular Ca2+ stores as judged by EGTA treatment. In additional experiments, isolation of stable clones derived from transfection of the CD45 S/A mutant into CD45- H45.01 cells showed sustained calcium flux after TCR engagement. The sustained calcium flux returned to baseline levels after addition of EGTA, suggesting that the expression of the CD45 S/A mutant may have prevented deactivation of plasma membrane calcium channels. Consideration of both transient and stable transfection systems suggests that in addition to being essential for initial events in T cell triggering, the intact CD45 D2, 19-aa insert is necessary for regulation of TCR-mediated calcium signaling pathways. PMID- 10679095 TI - Identification of a gene coding for a new squamous cell carcinoma antigen recognized by the CTL. AB - Peptide-based specific immunotherapy has resulted in tumor regression in some melanoma patients. However, tumor Ags and peptides for specific immunotherapy, except for treatment of melanomas, have not yet been well identified. In this study, we report a gene encoding a new squamous cell carcinoma (SCC) Ag recognized by cells of the HLA-A24-restricted and tumor-specific CTL line. This gene with 3958-bp length was transcribed from the chromosome 6q22 with six exons, and its mRNA was ubiquitously expressed in both SCCs and normal tissues, and partly expressed in adenocarcinomas. The deduced 958-aa sequence encoded by this gene showed no similarity to any known amino acid sequences. This gene product had a characteristic of an endoplasmic reticulum-resident protein. A 100-kDa protein was detected in the vast majority of SCCs from various tissues, in majority of renal cell carcinomas and brain tumors, and in about one-third of melanomas and adenocarcinomas from various organs other than the breast. In contrast, it was not expressed at all in any of the normal cells or tissues tested, including the testis and fetal liver. Three different peptides at positions 93-101, 161-169, and 899-907 of this Ag were recognized by this CTL line, and all of them induced HLA-A24-restricted and tumor-specific CTLs from PBMCs of SCC patients. Therefore, these peptides may be useful for peptide-based specific immunotherapy of HLA-A24+ patients with SCC in various organs, as well as for treatment of other cancer. PMID- 10679096 TI - IL-12-independent Th1-type immune responses to respiratory viral infection: requirement of IL-18 for IFN-gamma release in the lung but not for the differentiation of viral-reactive Th1-type lymphocytes. AB - We demonstrated that IL-12 was induced during primary or secondary pulmonary adenoviral infection in wild-type (wt) mice. However, cellular responses were not compromised in the lungs of IL-12-/- mice. The level of IFN-gamma in the lung was similar in wt and IL-12-/- mice during pulmonary viral infection. Upon Ag stimulation in vitro, lymphocytes from draining lymph nodes or spleen of infected IL-12-/- mice released large amounts of IFN-gamma, but not IL-4, which were comparable to those released by wt lymphocytes. Furthermore, a predominantly IgG2a response to adenoviral infection was unimpaired in IL-12-/- mice. These significant anti-adenoviral Th1-type responses in IL-12-/- mice led to an efficient clearance of virus-infected cells in the lung. Whether IL-18 was involved in IL-12-independent anti-adenoviral immune responses was investigated. Abrogation of endogenous IL-18 by an Ab resulted in diminished IFN-gamma release and lymphocytic infiltrate in the lung during adenoviral infection. Nevertheless, the development of lymphocytes of the Th1 phenotype was unimpaired in the absence of both IL-12 and IL-18. In contrast to their intact ability to mount Th1-type responses to viral infection, IL-12-/- mice suffered impaired Th1-type immune responses to pulmonary mycobacterial infection. Our findings suggest that IL-12, although induced, is not required for Th1-type responses to respiratory viral infection, in contrast to mycobacterial infection. IL-18 is required for the optimal release of IFN-gamma in the lung during viral infection, but is not required for the generation of virus-reactive Th1-type lymphocytes. Th1 differentiation during respiratory adenoviral infection may involve molecules different from IL-12 or IL-18. PMID- 10679097 TI - Schistosome infection of transgenic mice defines distinct and contrasting pathogenic roles for IL-4 and IL-13: IL-13 is a profibrotic agent. AB - Experimental Schistosoma mansoni infections of mice lead to a dynamic type 2 cytokine-mediated pathological process. We have used IL-4-deficient, IL-13 deficient, and IL-4/13-deficient mice to dissect the role of these cytokines in the development of immune response and pathology following S. mansoni infection. We demonstrate that while both of these cytokines are necessary to develop a robust Th2 cell-driven, eosinophil-rich granuloma response, they also perform disparate functions that identify novel sites for therapeutic intervention. IL-13 deficient mice demonstrated significantly enhanced survival following infection, which correlated with reduced hepatic fibrosis. In contrast, increased mortality was manifest in IL-4-deficient and IL-4/13-deficient mice, and this correlated with hepatocyte damage and intestinal pathology. Therefore, we demonstrate that during a dynamic type 2 cytokine disease process IL-13 is detrimental to survival following infection, whereas IL-4 is beneficial. PMID- 10679098 TI - Lipopolysaccharide inhibits HIV-1 infection of monocyte- derived macrophages through direct and sustained down-regulation of CC chemokine receptor 5. AB - It is now well established that HIV-1 requires interactions with both CD4 and a chemokine receptor on the host cell surface for efficient infection. The expression of the CCR5 chemokine receptor in human macrophages facilitates HIV-1 entry into these cells, which are considered important in HIV pathogenesis not only as viral reservoirs but also as modulators of altered inflammatory function in HIV disease and AIDS. LPS, a principal constituent of Gram-negative bacterial cell walls, is a potent stimulator of macrophages and has been shown to inhibit HIV infection in this population. We now present evidence that one mechanism by which LPS mediates its inhibitory effect on HIV-1 infection is through a direct and unusually sustained down-regulation of cell-surface CCR5 expression. This LPS mediated down-regulation of CCR5 expression was independent of de novo protein synthesis and differed from the rapid turnover of these chemokine receptors observed in response to two natural ligands, macrophage-inflammatory protein 1alpha and -1beta. LPS did not act by down-regulating CCR5 mRNA (mRNA levels actually increased slightly after LPS treatment) or by enhancing the degradation of internalized receptor. Rather, the observed failure of LPS-treated macrophages to rapidly restore CCR5 expression at the cell-surface appeared to result from altered recycling of chemokine receptors. Taken together, our results suggest a novel pathway of CCR5 recycling in LPS-stimulated human macrophages that might be targeted to control HIV-1 infection. PMID- 10679099 TI - Granulocyte-macrophage colony-stimulating factor in the innate immune response to Pneumocystis carinii pneumonia in mice. AB - Innate immunity plays an important role in pulmonary host defense against Pneumocystis carinii, an important pathogen in individuals with impaired cell mediated immunity. We investigated the role of GM-CSF in host defense in a model of P. carinii pneumonia induced by intratracheal inoculation of CD4-depleted mice. Lung GM-CSF levels increased progressively during the infection and were significantly greater than those in uninfected controls 3, 4, and 5 wk after inoculation. When GM-CSF gene-targeted mice (GM-/-) depleted of CD4+ cells were inoculated with P. carinii, the intensities of infection and inflammation were increased significantly compared with those in CD4-depleted wild-type mice. In contrast, transgenic expression of GM-CSF directed solely in the lungs of GM-/- mice (using the surfactant protein C promoter) dramatically decreased the intensity of infection and inflammation 4 wk after inoculation. The concentrations of surfactant proteins A and D were greater in both uninfected and infected GM-/- mice compared with those in wild-type controls, suggesting that this component of the innate response was preserved in the GM-/- mice. However, alveolar macrophages (AM) from GM-/- mice demonstrated impaired phagocytosis of purified murine P. carinii organisms in vitro compared with AM from wild-type mice. Similarly, AM production of TNF-alpha in response to P. carinii in vitro was totally absent in AM from GM-/- mice, while GM-CSF-replete mice produced abundant TNF in this setting. Thus, GM-CSF plays a critical role in the inflammatory response to P. carinii in the setting of impaired cell-mediated immunity through effects on AM activation. PMID- 10679100 TI - Purification and characterization of two mannan-binding lectins from mouse serum. AB - Mannan-binding lectin (MBL) is a serum protein that activates the complement system after binding to glycoconjugates found on the surface of microorganisms. By molecular cloning two forms of MBL have been identified in the mouse (mMBL-A and mMBL-C), but only mMBL-A has been purified and characterized at the protein level. MBL-C has been termed the liver form of MBL. The present report describes the purification and characterization of mMBL-A and mMBL-C from serum. The two forms of mMBL could be separated both by ion-exchange and carbohydrate-affinity chromatography. The initial identification by immunochemical technique was confirmed by N-terminal amino-acid sequencing. Both proteins give bands corresponding to polypeptide chains of 28 kDa on SDS-PAGE in the reduced state, but mMBL-A migrated more rapidly than mMBL-C in acid/urea-PAGE, in accordance with the calculated pIs. Both forms mediated activation of complement component C4 in mannan-coated microtiter wells. MBL-A showed a higher affinity for d glucose and alpha-methyl-d -glucose then did MBL-C. Serum concentrations of mMBL A in laboratory strains and wild mice were found to vary from 5 to 80 microg/ml, with wild mice tending to show higher levels than laboratory strains. PMID- 10679101 TI - Mice bearing late-stage tumors have normal functional systemic T cell responses in vitro and in vivo. AB - Immune suppression in tumor-bearing hosts is considered to be one factor causally associated with the growth of antigenic tumors. Support for this hypothesis has come from reports that spleen T cells in tumor-bearing mice are deficient in either priming or effector phase functions. We have reexamined this hypothesis in detail using multiple murine tumor models, including transplantable adenocarcinoma, melanoma, sarcoma, and thymoma, and also a transgenic model of spontaneous breast carcinoma. In both in vitro and in vivo assays of T cell function (proliferation, cytokine production, induction of CD8+ alloreactive CTL, and development of anti-keyhole limpet hemocyanin CD4+ T cells, rejection of allogeneic or syngeneic regressor tumors, respectively) we show that mice bearing sizable tumor burdens are not systemically suppressed and do not have diminished T cell functions. Therefore, if immune suppression is a causal function in the growth of antigenic tumor, the basis for escape from immune destruction is likely to be dependent upon tumor-induced T cell dysfunction at the site of tumor growth. PMID- 10679102 TI - Decreased resistance of B cell-deficient mice to infection with Toxoplasma gondii despite unimpaired expression of IFN-gamma, TNF-alpha, and inducible nitric oxide synthase. AB - The role of B cells in resistance against Toxoplasma gondii was studied using B cell-deficient (muMT) mice. Following peroral infection with 10 cysts of the ME49 strain, all muMT mice survived the acute stage of the infection but died between 3 and 4 wk after infection. In contrast, all control mice were alive at 8 wk after infection. At the stage during which muMT animals succumbed to the infection, parasite replication and pathology were most evident in their brains; small numbers of tachyzoites were also detectable in their lungs. Significantly greater numbers of T. gondii cysts and areas of inflammation associated with tachyzoites were observed in brains of muMT than in control mice. Large areas of necrosis associated with numerous tachyzoites were observed only in brains of muMT mice. Anti-T. gondii IgG Abs were detected only in sera of control mice, whereas similar levels of IFN-gamma were detected in sera of both strains of mice. Amounts of mRNA for IFN-gamma, IL-10, and inducible NO synthase in the brain did not differ between infected muMT and control mice. Expression of mRNA for TNF-alpha was increased in brains of muMT mice. Administration of polyclonal rabbit anti-T. gondii IgG Ab prevented early mortality and pathology associated with tachyzoites in the brain in the infected muMT mice. These results indicate that B cells play an important role, most likely through their production of specific Abs, in resistance to persistent active (tachyzoite) infection with T. gondii in mice, especially in the brain and lung. PMID- 10679103 TI - Th cell-deficient mice control influenza virus infection more effectively than Th and B cell-deficient mice: evidence for a Th-independent contribution by B cells to virus clearance. AB - The notion that MHC class I- restricted CD8+ T (Tc) cells are capable of resolving autonomously infections with influenza virus is based largely on studies testing virus strains of low pathogenicity in CD4+ T (Th) cell deficient/depleted mice. To test whether this holds also for pathogenic strains and to exclude possible contributions by B cells, we analyzed PR8 infection in Th cell-depleted B cell-deficient (muMT) mice. These mice, termed muMT (-CD4), showed 80% mortality after infection with a small dose of PR8, which resulted in insignificant mortality in intact or Th cell-depleted BALB/c mice. Infection of muMT(-CD4) mice with a virus of low pathogenicity was resolved without mortality, but, compared with intact BALB/c mice, with delay of approximately 5 and approximately 20 days from lung and nose, respectively. The low mortality of Th cell-depleted BALB/c mice suggested that B cells contributed to recovery in a Th independent manner. This was verified by showing that transfer of 8-10 million T cell-depleted naive spleen cells into muMT(-CD4) mice 1 day before infection reduced mortality to 0%. The mechanism by which B cells improved recovery was investigated. We found no evidence that they operated by improving the lung associated Tc response. Treatment of infected muMT(-CD4) mice with normal mouse serum spiked with hemagglutinin-specific IgM did not reduce mortality. Taken together, the data show that 1) the Tc response is capable of resolving autonomously (in conjunction with innate defenses) influenza virus infections, although with substantial delay compared with intact mice, and 2) B cells can contribute to recovery by a Th-independent mechanism. PMID- 10679104 TI - Neutralization of IL-18 reduces neutrophil tissue accumulation and protects mice against lethal Escherichia coli and Salmonella typhimurium endotoxemia. AB - In addition to stimulating IFN-gamma synthesis, IL-18 also possesses inflammatory effects by inducing synthesis of the proinflammatory cytokines TNF and IL-1beta and the chemokines IL-8 and macrophage inflammatory protein-1alpha. We hypothesized that neutralization of IL-18 would have a beneficial effect in lethal endotoxemia in mice. IL-1beta converting enzyme (ICE)-deficient mice, lacking the ability to process mature IL-18 and IL-1beta, were completely resistant to lethal endotoxemia induced by LPS derived from either Escherichia coli or Salmonella typhimurium. In contrast, both wild-type and IL-1beta-/- mice were equally susceptible to the lethal effects of LPS, implicating that absence of mature IL-18 or IFN-gamma but not IL-1beta in ICE-/- mice is responsible for this resistance. However, IFN-gamma-deficient mice were not resistant to S. typhimurium LPS, suggesting an IFN-gamma-independent role for IL-18. Anti-IL-18 Abs protected mice against a lethal injection of either LPS. Anti-IL-18 treatment also reduced neutrophil accumulation in liver and lungs. The increased survival was accompanied by decreased levels of IFN-gamma and macrophage inflammatory protein-2 in anti-IL-18-treated animals challenged with E. coli LPS, whereas IFN gamma and TNF concentrations were decreased in treated mice challenged with S. typhimurium. In conclusion, neutralization of IL-18 during lethal endotoxemia protects mice against lethal effects of LPS. This protection is partly mediated through inhibition of IFN-gamma production, but mechanisms involving decreased neutrophil-mediated tissue damage due to the reduction of either chemokines (E. coli LPS) or TNF (S. typhimurium LPS) synthesis by anti-IL-18 treatment may also be involved. PMID- 10679105 TI - Role of CC chemokines (macrophage inflammatory protein-1 beta, monocyte chemoattractant protein-1, RANTES) in acute lung injury in rats. AB - The role of the CC chemokines, macrophage inflammatory protein-1 beta (MIP-1 beta), monocyte chemotactic peptide-1 (MCP-1), and RANTES, in acute lung inflammatory injury induced by intrapulmonary deposition of IgG immune complexes injury in rats was determined. Rat MIP-1 beta, MCP-1, and RANTES were cloned, the proteins were expressed, and neutralizing Abs were developed. mRNA and protein expression for MIP-1 beta and MCP-1 were up-regulated during the inflammatory response, while mRNA and protein expression for RANTES were constitutive and unchanged during the inflammatory response. Treatment of rats with anti-MIP-1 beta Ab significantly decreased vascular permeability by 37% (p = 0.012), reduced neutrophil recruitment into lung by 65% (p = 0.047), and suppressed levels of TNF alpha in bronchoalveolar lavage fluids by 61% (p = 0.008). Treatment of rats with anti-rat MCP-1 or anti-rat RANTES had no effect on the development of lung injury. In animals pretreated intratracheally with blocking Abs to MCP-1, RANTES, or MIP-1 beta, significant reductions in the bronchoalveolar lavage content of these chemokines occurred, suggesting that these Abs had reached their targets. Conversely, exogenously MIP-1 beta, but not RANTES or MCP-1, caused enhancement of the lung vascular leak. These data indicate that MIP-1 beta, but not MCP-1 or RANTES, plays an important role in intrapulmonary recruitment of neutrophils and development of lung injury in the model employed. The findings suggest that in chemokine-dependent inflammatory responses in lung CC chemokines do not necessarily demonstrate redundant function. PMID- 10679106 TI - Lipoxin A4 inhibits IL-1 beta-induced IL-6, IL-8, and matrix metalloproteinase-3 production in human synovial fibroblasts and enhances synthesis of tissue inhibitors of metalloproteinases. AB - Lipoxins are a novel class of endogenous eicosanoid mediators that potently inhibit inflammatory events by signaling via specific receptors expressed on phagocytic cells. Animal models have shown that lipoxin A4 (LXA4) down-regulates inflammation in vivo. Here we demonstrate, for the first time, the expression of LXA4 receptors, and their up-regulation by IL-1 beta, in normal human synovial fibroblasts (SF). We examined whether exogenous LXA4 abrogated IL-1 beta stimulation of SF in vitro. IL-1 beta induced the synthesis of IL-6, IL-8, and matrix metalloproteinases (MMP)-1 and -3. At nanomolar concentrations, LXA4 inhibited these IL-1 beta responses with reduction of IL-6 and IL-8 synthesis, by 45 +/- 7% and 75 +/- 11%, respectively, and prevented IL-1 beta-induced MMP-3 synthesis without significantly affecting MMP-1 levels. Furthermore, LXA4 induced a 2-fold increase of tissue inhibitor of metalloproteinase (TIMP)-1 and a approximately 3-fold increase of TIMP-2 protein levels. LXA4 inhibitory responses were dose dependent and were abrogated by pretreatment with LXA4 receptor antiserum. LXA4-induced changes of IL-6 and TIMP were accompanied by parallel changes in mRNA levels. These results indicate that LXA4 in activated SF inhibits the synthesis of inflammatory cytokines and MMP and stimulates TIMP production in vitro. These findings suggest that LXA4 may be involved in a negative feedback loop opposing inflammatory cytokine-induced activation of SF. PMID- 10679107 TI - Role for IgE in airway secretions: IgE immune complexes are more potent inducers than antigen alone of airway inflammation in a murine model. AB - IgE is present in airway secretions from human patients with allergic rhinitis and bronchial asthma. However, the contribution of IgE present locally to the overall airway inflammation is not well understood. We hypothesize that Ag specific IgE can capture airborne Ags and form immune complexes. These immune complexes may function as potent inducers of immune responses in the lung, contributing to the perpetuation of airway inflammation. BALB/c mice were first sensitized with OVA in alum systemically and then challenged with nebulized OVA. Bronchoalveolar lavage (BAL) fluid from these mice contained significant amounts of IgE, of which >50% was Ag specific. The IgE levels in airway secretions remained elevated for more than 15 days after the termination of Ag exposure. Significant amounts of IgE-OVA immune complexes were detected in BAL fluid from the OVA-challenged mice. For comparison of IgE immune complexes vs Ag alone, we treated OVA-immunized mice with intranasal administration of trinitrophenyl-OVA or trinitrophenyl-OVA-anti-DNP IgE. Those treated with the immune complexes showed significantly higher levels of IL-4 and more pronounced eosinophilia in BAL fluid than did those receiving the Ag alone. The IgE immune complexes did not augment the inflammatory response in high affinity IgE receptor (FcepsilonRI) deficient mice. We conclude that IgE present in the airways can capture the Ag and that the immune complexes thus formed may augment allergic airway response in an FcepsilonRI-dependent manner. Thus, IgE present in airway secretions may facilitate Ag-mediated allergic airway inflammation. PMID- 10679108 TI - Neutrophil activation by bacterial lipoprotein versus lipopolysaccharide: differential requirements for serum and CD14. AB - Neutrophil activation plays an important role in the inflammatory response to Gram-negative bacterial infections. LPS has been shown to be a major mediator of neutrophil activation which is accompanied by an early down-regulation of L selectin and up-regulation of CD1lb/CD18. In this study, we investigated whether lipoprotein (LP), the most abundant protein in the outer membrane of bacteria from the family Enterobacteriaceae, can activate neutrophils and whether this activation is mediated by mechanisms that differ from those used by LPS or Escherichia coli diphosphoryl lipid A (EcDPLA). Neutrophil activation was assessed by measuring down-regulation of L-selectin and up-regulation of CD11b/CD18. When comparing molar concentrations of LP vs EcDPLA, LP was more potent (four times) at activating neutrophils. In contrast to LPS/EcDPLA, LP activation of neutrophils was serum independent. However, LP activation of neutrophils was enhanced by the addition of soluble CD14 and/or LPS-binding protein. In the presence of serum, LP activation of neutrophils was inhibited by different mAbs to CD14. This inhibition was significantly reduced or absent when performed in the absence of serum. Diphosphoryl lipid A from Rhodobacter spheroides (RaDPLA) completely inhibited LPS/EcDPLA activation of neutrophils but only slightly inhibited LP activation of neutrophils. These results suggest that LP activation of human neutrophils can be mediated by a mechanism that is different from LPS activation and that LP is a potentially important component in the development of diseases caused by Gram-negative bacteria of the family Enterobacteriaceae. PMID- 10679109 TI - Functional genomic analysis in arthritis-affected cartilage: yin-yang regulation of inflammatory mediators by alpha 5 beta 1 and alpha V beta 3 integrins. AB - Osteoarthritis-affected cartilage exhibits enhanced expression of fibronectin (FN) and osteopontin (OPN) mRNA in differential display and bioinformatics screen. Functional genomic analysis shows that the engagement of the integrin receptors alpha 5 beta 1 and alpha v beta 3 of FN and OPN, respectively, have profound effects on chondrocyte functions. Ligation of alpha 5 beta 1 using activating mAb JBS5 (which acts as agonist similar to FN N-terminal fragment) up regulates the inflammatory mediators such as NO and PGE2 as well as the cytokines, IL-6 and IL-8. Furthermore, up-regulation of these proinflammatory mediators by alpha 5 beta1 integrin ligation is mediated via induction and autocrine production of IL-1 beta, because type II soluble IL-1 decoy receptor inhibits their production. In contrast, alpha v beta 3 complex-specific function blocking mAb (LM609), which acts as an agonist similar to OPN, attenuates the production of IL-1 beta, NO, and PGE2 (triggered by alpha 5 beta 1, IL-1 beta, IL 18, or IL-1 beta, TNF-alpha, plus LPS) in a dominant negative fashion by osteoarthritis-affected cartilage and activated bovine chondrocytes. These data demonstrate a cross-talk in signaling mechanisms among integrins and show that integrin-mediated "outside in" and "inside out" signaling very likely influences cartilage homeostasis, and its deregulation may play a role in the pathogenesis of osteoarthritis. PMID- 10679110 TI - Lipopolysaccharide induces scavenger receptor A expression in mouse macrophages: a divergent response relative to human THP-1 monocyte/macrophages. AB - Gene deletion studies indicate that the macrophage scavenger receptor A (SR-A) protects mice from LPS-induced endotoxemia. Paradoxically, cultured human monocyte-derived macrophages down-regulate SR-A expression when exposed to LPS. We found that human THP-1 monocyte/macrophages decrease SR-A expression in response to LPS independent of their differentiation status. In contrast, primary and elicited mouse peritoneal macrophages as well as the J774A.1 and RAW264.7 mouse macrophage lines increase SR-A expression in response to LPS. Exposure to LPS caused J774A.1 and RAW264.7 cells to increase SR-A transcripts by 3- and 5 fold, respectively. LPS caused a concomitant 3-fold increase in SR-A protein levels and increased cell membrane expression of the receptor. RAW264.7 cells increased SR-A transcript levels in response to LPS at concentrations as low as 1 ng/ml, and the response was saturated at 10 ng/ml. The LPS induction of SR-A transcripts required continual protein synthesis and began at 8 h, peaked by 16 h, and persisted for at least 48 h. LPS induction did not increase SR-A gene transcription or affect alternative transcript splicing, but mildly increased mature transcript stability and proceeded in the presence of actinomycin D. Finally, treatment of RAW264.7 cells with TNF-alpha did not induce SR-A transcript levels, indicating that a TNF-alpha autocrine/paracrine signaling mechanism alone is not sufficient to recapitulate the LPS induction of SR-A transcripts. The induction of SR-A expression by LPS-stimulated mouse macrophages is the opposite of the down-regulation of SR-A reported in human monocyte-derived macrophages and may have implications for the observed resistance mice show toward endotoxemia. PMID- 10679111 TI - IFN-gamma-inducing factor (IL-18) increases allergic sensitization, serum IgE, Th2 cytokines, and airway eosinophilia in a mouse model of allergic asthma. AB - We investigated the effects of IFN-gamma-inducing factor (IL-18) in a ragweed (RW) mouse model of allergic asthma. Administration of IL-18 in conjunction with allergic sensitization and challenge in wild-type, but not IFN-gamma -/- mice, inhibited the bronchoalveolar lavage (BAL) eosinophilia induced by RW challenge, and increased serum levels of RW-specific IgG2a and production of IFN-gamma from splenocytes cultured with RW, indicating a critical role for IFN-gamma in mediating these effects. Paradoxically, the same treatment schedule in WT mice increased serum levels of RW-specific IgE and IgG1, and production of IL-4 and IL 5 from splenocytes cultured with RW. When the effects of the same IL-18 treatment schedule were allowed to mature for 3 wk, the inhibition of lung eosinophil recruitment was replaced by augmentation of lung eosinophil recruitment. In another experiment, IL-18 administered only with allergic sensitization increased BAL eosinophilia and lung expression of IL-5 and IFN-gamma, while IL-18 administered only with RW challenge decreased BAL eosinophilia and increased lung IFN-gamma expression, while lung expression of IL-5 remained unchanged. IL-18 administered without RW or adjuvant to naive mice increased total serum IgE levels. Finally, intrapulmonary administrations of IL-18 plus RW in naive mice dramatically increased Th2 cytokine production, IgE levels, eosinophil recruitment, and airway mucus, demonstrating induction of allergic sensitization. This is the first report demonstrating that IL-18 promotes a Th2 phenotype in vivo, and potently induces allergic sensitization. These results suggest that IL 18 may contribute to the pathogenesis of allergic asthma. PMID- 10679112 TI - A conserved mycobacterial heat shock protein (hsp) 70 sequence prevents adjuvant arthritis upon nasal administration and induces IL-10-producing T cells that cross-react with the mammalian self-hsp70 homologue. AB - Immunization with Mycobacterium tuberculosis heat shock protein (hsp) 60 has been shown to protect rats from experimental arthritis. Previously, the protection inducing capacity was shown to reside in the evolutionary conserved parts of the molecule. Now we have studied the nature of the arthritis suppressive capacity of a distinct, antigenically unrelated protein, M. tuberculosis hsp70. Again, a conserved mycobacterial hsp70 sequence was found to be immunogenic and to induce T cells that cross-reacted with the rat homologue sequence. However, in this case parenteral immunization with the peptide containing the critical cross-reactive T cell epitope did not suppress disease. Upon analysis of cytokines produced by these peptide-specific T cells, high IL-10 production was found, as was the case with T cells responsive to whole hsp70 protein. Nasal administration of this peptide was found to lead to inhibition of subsequent adjuvant arthritis induction. The data presented here shows the intrinsic capacity of conserved bacterial hsp to trigger self-hsp cross-reactive T cells with the potential to down-regulate arthritis via IL-10. PMID- 10679113 TI - T lymphocytes activated by persistent viral infection differentially modify the expression of metalloproteinases and their endogenous inhibitors, TIMPs, in human astrocytes: relevance to HTLV-I-induced neurological disease. AB - Activation of T lymphocytes by human pathogens is a key step in the development of immune-mediated neurologic diseases. Because of their ability to invade the CNS and their increased secretion of proinflammatory cytokines, activated CD4+ T cells are thought to play a crucial role in pathogenesis. In the present study, we examined the expression of inflammatory mediators the cytokine-induced metalloproteinases (MMP-2, -3, and -9) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMP-1, -2, and -3), in human astrocytes in response to activated T cells. We used a model system of CD4+ T lymphocytes activated by persistent viral infection (human T lymphotropic virus, HTLV-I) in transient contact with human astrocytes. Interaction with T cells resulted in increased production of MMP-3 and active MMP-9 in astrocytes despite increased expression of endogenous inhibitors, TIMP-1 and TIMP-3. These data suggest perturbation of the MMP/TIMP balance. These changes in MMP and TIMP expression were mediated, in part, by soluble factors (presumably cytokines) secreted by activated T cells. Integrin-mediated cell adhesion is also involved in the change in MMP level, since blockade of integrin subunits (alpha1, alpha3, alpha5, and beta1) on T cells resulted in less astrocytic MMP-9-induced expression. Interestingly, in CNS tissues from neurological HTLV-I-infected patients, MMP-9 was detected in neural cells within the perivascular space, which is infiltrated by mononuclear cells. Altogether, these data emphasize the importance of the MMP TIMP axis in the complex interaction between the CNS and invading immune cells in the context of virally mediated T cell activation. PMID- 10679114 TI - Induction of chemokine secretion and enhancement of contact-dependent macrophage cytotoxicity by engineered expression of granulocyte-macrophage colony stimulating factor in human colon cancer cells. AB - We investigated the role of tumor cell-derived GM-CSF in recruitment and tumoricidal activation of tissue macrophages. Transfection of the murine GM-CSF gene into KM12SM human colon cancer cells decreased the tumorigenicity of transfected cells and nontransfected bystander colon cancer cells in nude mice. Sequential tissue sections from sites injected with high GM-CSF-producing tumor cells (but not from nontransfected or low GM-CSF-producing cells) demonstrated a dense infiltration of polymorphonuclear cells (PMN), followed by infiltration of macrophages, which correlated with expression of the macrophage-inflammatory protein-1alpha and the monocyte chemoattractant protein-1 (MCP-1) in mouse PMN and macrophages. GM-CSF-producing KM12SM cells were highly sensitive to lysis by mouse macrophages and also increased macrophage-mediated lysis of bystander nontransfected KM12SM cells. The incubation of macrophages with GM-CSF induced expression of the CD11b surface adhesion molecule, which was associated with increased attachment to tumor cells. All KM12SM cells were sensitive to macrophage-mediated lysis in the presence of rGM-CSF and recombinant MCP-1. Collectively, the results demonstrate that tumor cell-derived GM-CSF stimulates PMN and macrophages to secrete macrophage-inflammatory protein-1alpha and MCP-1, which triggers recruitment of mononuclear cells, induces expression of adhesion molecules on macrophages, and enhances contact-dependent cytolysis of tumor cells. PMID- 10679115 TI - Expression and contribution of endogenous IL-13 in an experimental model of sepsis. AB - IL-13 has been shown to exert potent anti-inflammatory properties. In this study, we elucidated the functional role of endogenous IL-13 in a murine model of septic peritonitis induced by cecal ligation and puncture (CLP). Initial studies demonstrated that the level of IL-13 increased in tissues including liver, lung, and kidney, whereas no considerable increase was found in either peritoneal fluid or serum after CLP. Immunohistochemically, IL-13-positive cells were Kupffer cells in liver, alveolar macrophages in lung, and epithelial cells of urinary tubules in kidney. IL-13 blockade with anti-IL-13 Abs significantly decreased the survival rate of mice after CLP from 53% to 14% on day 7 compared with control. To determine the potential mechanisms whereby IL-13 exerted a protective role in this model, the effects of anti-IL-13 Abs on both local and systemic inflammation were investigated. Administration of anti-IL-13 Abs did not alter the leukocyte infiltration and bacterial load in the peritoneum after CLP but dramatically increased the neutrophil influx in tissues after CLP, an effect that was accompanied by significant increases in the serum levels of aspartate transaminase, alanine transaminase, blood urea nitrogen, and creatinine. Tissue injury caused by IL-13 blockade was associated with increases in mRNA and the protein levels of CXC chemokines macrophage inflammatory protein-2 and KC as well as the CC chemokine macrophage inflammatory protein-1alpha and the proinflammatory cytokine TNF-alpha. Collectively, these results suggest that endogenous IL-13 protected mice from CLP-induced lethality by modulating inflammatory responses via suppression of overzealous production of inflammatory cytokines/chemokines in tissues. PMID- 10679116 TI - Pivotal role of CCR1-positive leukocytes in bleomycin-induced lung fibrosis in mice. AB - We have investigated the involvement of chemokine receptor CCR1-positive cells in bleomycin-induced lung injury, a model of pulmonary fibrosis. After bleomycin challenge in C57BL/6J mice, the expression of CCR1 mRNA increased and peaked at day 7, which paralleled to the expression of its ligands, macrophage-inflammatory protein-1 alpha and RANTES. Immunohistochemical study showed that CCR1-positive cells accumulated in the interstitial inflammatory site. Furthermore, the treatment of anti-CCR1 Ab significantly reduced the accumulation of inflammatory cells and collagen deposition, resulting in dramatic improvement of survival. These results suggest that CCR1-positive cells play significant roles in the pathogenesis of pulmonary fibrosis subsequent to bleomycin-induced lung injury, and that CCR1 could be a novel molecular target for intervention therapy against pulmonary fibrosis. PMID- 10679117 TI - Time-dependent loss of Mac-1 from infiltrating neutrophils in the reperfused myocardium. AB - Numerous studies have shown that polymorphonuclear neutrophils (PMNs) infiltrate the myocardium immediately after reperfusion of infarcted tissue. Studies with mAbs in vivo and cellular studies in vitro suggest that PMN-induced injury of the cardiac myocyte involve Mac-1 adhesion to myocyte ICAM-1. In this study we demonstrate that PMNs that have infiltrated the ischemic area begin to lose Mac-1 within the first 3 h. By the fifth hour of reperfusion, minimal CD11b staining is seen on PMNs using immunostaining, whereas CD11a remained unchanged. Immunoreactivity of postreperfusion cardiac lymph with R15.7 (anti-CD18) or MY904 (anti-CD11b) was positive in all animals but not for CD11a (R7.1), indicating a specific loss of Mac-1. Immunoprecipitation with either R15.7 or MY904 resulted in identical peptides (a doublet at 190 kDa and a band at 80 kDa), suggesting that both alpha and beta subunits of Mac-1 heterodimer were released. Immunoprecipitation of control PMN lysates revealed bands of 198 kDa and 91 kDa slightly greater than those from the released Mac-1. An in vitro model of homotypic aggregation showed a similar loss of Mac-1 from PMNs; immunoprecipitates of the supernatant demonstrated peptide bands identical with those found in postischemic cardiac lymph. The appearance of soluble Mac-1 in vitro was prevented by anti-CD18 mAb, R15.7, and also by protease inhibition by PMSF. Thus, in vivo and in vitro, activated PMNs lose Mac-1 in a process that may be dependent upon adhesion and subsequent proteolysis. PMID- 10679118 TI - IFN-gamma shapes immune invasion of the central nervous system via regulation of chemokines. AB - Dynamic interplay between cytokines and chemokines directs trafficking of leukocyte subpopulations to tissues in autoimmune inflammation. We have examined the role of IFN-gamma in directing chemokine production and leukocyte infiltration to the CNS in experimental autoimmune encephalomyelitis (EAE). BALB/c and C57BL/6 mice are resistant to induction of EAE by immunization with myelin basic protein. However, IFN-gamma-deficient (BALB/c) and IFN-gammaR deficient (C57BL/6) mice developed rapidly progressing lethal disease. Widespread demyelination and disseminated leukocytic infiltration of spinal cord were seen, unlike the focal perivascular infiltrates in SJL/J mice. Gr-1+ neutrophils predominated in CNS, and CD4+ T cells with an activated (CD69+, CD25+) phenotype and eosinophils were also present. RANTES and macrophage chemoattractant protein 1, normally up-regulated in EAE, were undetectable in IFN-gamma- and IFN-gammaR deficient mice. Macrophage inflammatory protein-2 and T cell activation gene-3, both neutrophil-attracting chemokines, were strongly up-regulated. There was no induction of the Th2 cytokines, IL-4, IL-10, or IL-13. RNase protection assays and RT-PCR showed the prevalence of IL-2, IL-3, and IL-15, but no increase in IL 12p40 mRNA levels in IFN-gamma- or IFN-gammaR-deficient mice with EAE. Lymph node cells from IFN-gamma-deficient mice proliferated in response to myelin basic protein, whereas BALB/c lymph node cells did not. These findings show a regulatory role for IFN-gamma in EAE, acting on T cell proliferation and directing chemokine production, with profound implications for the onset and progression of disease. PMID- 10679119 TI - IL-8 reduced tumorigenicity of human ovarian cancer in vivo due to neutrophil infiltration. AB - Paclitaxel is a frontline therapy for ovarian cancer. Our laboratory has shown that paclitaxel induces IL-8, a member of the C-X-C family of chemokines, in subsets of human ovarian cancer cells. However, the critical issue concerns the biological significance of this chemokine in human ovarian cancer. To study the influence of IL-8 on tumor growth, human ovarian cancer cell lines were transfected with an expression vector for human IL-8 and tested for their ability to form tumors in nude mice. IL-8 expression by the transfected cells did not alter their growth properties in vitro. In contrast, tumor growth in vivo was significantly attenuated in animals receiving IL-8-expressing cells when compared with mice injected with control cells. As additional evidence that IL-8 is a crucial factor in tumor growth, it was noted that ovarian cell lines in which constitutive IL-8 expression is elevated did not form tumors. Injection of neutralizing Ab to IL-8 reverted the phenotype and caused tumor growth in vivo. Examination of tissue from the inoculation site revealed a dramatically elevated cellularity, containing neutrophils and macrophages, in mice receiving IL-8 expressing tumor cells. These results suggest that IL-8 production by human ovarian tumor cells can play a role in reducing the rate of tumor growth; this effect may be mediated by the increased targeting of neutrophil and other mononuclear cells to the tumor injection site. These studies indicate a role for IL-8 in ovarian cancer control and suggest that chemotherapy-induced IL-8 may have a positive role in controlling tumor growth. PMID- 10679120 TI - Gene therapy for chronic relapsing experimental allergic encephalomyelitis using cells expressing a novel soluble p75 dimeric TNF receptor. AB - In a murine relapsing experimental allergic encephalomyelitis (EAE) model, gene therapy to block TNF was investigated with the use of a retroviral dimeric p75 TNF receptor (dTNFR) construct. To effectively produce these TNF inhibitors in vivo, a conditionally immortalized syngeneic fibroblast line was established, using a temperature-sensitive SV40 large T Ag-expressing retrovirus. These cells were subsequently infected with a retrovirus expressing soluble dTNFR. CNS injected cells could be detected 3 mo after transplantation and were shown to produce the transgene product by immunocytochemistry and ELISA of tissue fluids. These levels of dTNFR protein were biologically active and could significantly ameliorate both acute and relapsing EAE. This cell-based gene-vector approach is ideal for delivering proteins to the CNS and has particular relevance to the control of inflammatory CNS disease. PMID- 10679121 TI - Accumulation of clonally related B lymphocytes in the cerebrospinal fluid of multiple sclerosis patients. AB - The accumulation of B lymphocyte clones in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) and patients with other neurological disorders was investigated using PCR technologies. Oligoclonal B cell accumulations were detected in 10 of 10 MS patients, but only in 3 of 10 of the patients with other neurological disorders. Analyses of the Ig V(D)J sequences on the CSF from MS patients disclosed that VH3 and VH4 genes were extensively mutated compared with germline sequences. Moreover, a substantial proportion of the molecular clones analyzed shared the same third CDR of the H chain variable region gene (HCDR3) and the same VH genes, albeit with different numbers and locations of point mutations, thus indicating an ongoing process of intraclonal diversification. A larger number of clonally related VH sequences could be obtained by using a VH3 gene-specific PCR so that genealogical trees depicting the process of diversification could be drawn. Analyses of the Ig V(D)J from the CSF of a patient with viral meningitis and oligoclonal B cell accumulations revealed that VH3 genes were extensively mutated. However, no intraclonal diversification could be observed even using VH3 gene-specific PCR methodologies. Clone-specific PCR and sequencing was used to detect the V(D)J found in the CSF of one MS patient in the PBL of the same patient. Only 1/3 of the V(D)J sequences investigated could be demonstrated in the PBL, indicating that the V(D)J genes utilized by B cells in the CSF are much less represented in the PBL. Collectively, the data suggest that in MS there is a compartmentalized clonal expansion. PMID- 10679122 TI - Naturally developing memory T cell xenoreactivity to swine antigens in human peripheral blood lymphocytes. AB - Naturally developing xenospecific Abs are well-documented barriers to xenograft transplantation in humans, but whether analogous xenoreactive T cell immunity develops is not known. We used an enzyme-linked immunospot assay to determine the frequency and cytokine profiles of xenoreactive PBLs from a panel of human volunteers. Because naive T cells produce only IL-2 in short term culture, IFN gamma production by this approach is a measure of a memory immune response. Stimulation of human PBLs or purified T lymphocytes with stimulator cells from inbred swine revealed a high frequency of IFN-gamma producers with 5-fold fewer IL-2 producers. In contrast, lymphocytes obtained from neonatal umbilical cord blood contained swine-specific IL-2 producers but few IFN-gamma producers, which is what one would expect to find with a naive phenotype. Moreover, PBLs from adults with a history of abstention from pork consumption responded to swine cells with a significantly lower frequency of IFN-gamma producers than PBLs from adults with unrestricted diets did, suggesting that pork consumption may result in priming of swine-specific T cell immunity. Our findings provide the first evidence for naturally occurring xenospecific T cell immunity in humans. The detected strength of this memory response suggests that it will present a formidable barrier to transplantation of swine organs. PMID- 10679123 TI - Systemic activation and antigen-driven oligoclonal expansion of T cells in a mouse model of colitis. AB - Transfer of CD4+CD45RBhigh T cells into immunodeficient mice results in both the expansion of the transferred T cells and colitis. Here we show that colitis pathogenesis requires expression of MHC class II molecules by the immune deficient host. Analysis of the TCRbeta repertoire of the cells found in the large intestine of diseased mice revealed a population with restricted TCR diversity. Furthermore, nucleotide sequence analysis demonstrated the selection for particular CDR3beta amino acid sequence motifs. Collectively, these data indicate that the expansion of T cells in the intestine and colitis pathogenesis are likely to require the activation of Ag-specific T cells, as opposed to nonspecific or superantigen-mediated events. There is relatively little overlap, however, when the TCR repertoires of different individuals are compared, suggesting that a number of Ags can contribute to T cell expansion and the generation of a T cell population in the intestine. Surprisingly, many of the expanded clones found in the large intestine also were found in the spleen and elsewhere, although inflammation is localized to the colon. Additionally, donor derived T cells appear to be activated in both the intestine and the spleen at early time points after cell transfer. Together, these results strongly suggest that disease induction in this model involves either the early and systemic activation of antigen-specific T cells or the rapid dispersal of T cells activated at a particular site. PMID- 10679124 TI - The human UTY gene encodes a novel HLA-B8-restricted H-Y antigen. AB - The mammalian Y chromosome encodes male-specific minor histocompatibility (H-Y) Ags that are recognized by female T cells in an MHC-restricted manner. Two human H-Y epitopes presented by HLA-A2 and HLA-B7, respectively, have been identified previously and both are derived from the SMCY gene. We previously isolated CD8+ CTL clones that recognized a male-specific minor histocompatibility Ag presented by HLA-B8. In contrast to the SMCY-encoded H-Y epitopes, the B8/H-Y Ag was not presented by fibroblasts from male donors, suggesting that it was encoded by a novel gene. We now report that the HLA-B8-restricted H-Y epitope is defined by the octameric peptide LPHNHTDL corresponding to aa residues 566-573 of the human UTY protein. Transcription of the UTY gene is detected in a wide range of human tissues, but presentation of the UTY-derived H-Y epitope to CTL by cultured human cells shows significant cell-type specificity. Identification of this CTL-defined H-Y epitope should facilitate analysis of its contribution to graft/host interactions following sex-mismatched organ and bone marrow transplantation. PMID- 10679125 TI - Analysis and significance of anti-latent membrane protein-1 antibodies in the sera of patients with EBV-associated diseases. AB - Anti-latent membrane protein-1 (LMP-1) is an EBV-encoded type III integral membrane protein with oncogenic potential that is expressed most consistently in various EBV-associated malignancies. Unlike many other EBV proteins, LMP-1 Abs have rarely been demonstrated in EBV-associated disease conditions. We established a high level LMP-1-expressing cell clone and used it for the detection, quantitation, and characterization of these Abs in various human sera in immunoblots and ELISA. Our results demonstrate that, in contrast to the commonly held notion, LMP-1 induces significant humoral immune responses in EBV associated malignant conditions especially in nasopharyngeal carcinoma (NPC) patients in whom >70% sera are positive for these Abs, and their titers correlate with the clinical condition of the tumors. Interestingly, anti-LMP-1 Abs of IgA isotype were found only in NPC patients. These Abs were absent from the sera of infectious mononucleosis and chronic EBV infection patients, whereas a small fraction ( approximately 5%) of the healthy, EBV-seropositive individuals were positive for them; however, their OD values were much lower than those of NPC patients. These studies demonstrate, for the first time, the potential significance of LMP-1-specific Abs for the diagnosis and prognosis of EBV associated malignancies, especially of NPC. PMID- 10679126 TI - Accumulation of common T cell clonotypes in the salivary glands of patients with human T lymphotropic virus type I-associated and idiopathic Sjogren's syndrome. AB - To clarify the pathogenesis of human T lymphotropic virus type I (HTLV-I) associated Sjogren's syndrome (SS), the TCR Vbeta gene usage by the infiltrating lymphocytes in the target organ was examined. The Vbeta families predominantly used in the labial salivary gland (LSG) from the HTLV-I-seropositive (HTLV-I+) SS patients were more restricted than those from the HTLV-I-seronegative (idiopathic) SS patients, and were commonly Vbeta5.2, Vbeta6, and Vbeta7. The single-strand conformation polymorphism analysis revealed that T cell clonotypes with Vbeta5.2, Vbeta6, and Vbeta7 accumulate in the LSG from the HTLV-I+ and idiopathic SS patients. Among junctional sequences of the most dominant Vbeta7 transcripts, the conserved amino acid motif (QDXG: X is any amino acid) was found in six of the five HTLV-I+ SS patients and was also detected in two of the five idiopathic SS patients. Using the probes specific to the motif, the Vbeta7 transcripts with the motif were detected in the LSG from all of the seven HTLV-I+ and five of the six idiopathic SS patients, but not from eight healthy subjects. The Vbeta7 transcripts with this motif were also detected in the HTLV-I-infected T cell lines obtained from the LSG of an HTLV-I+ SS patient. The accumulation of HTLV-I-infected T cells expressing TCR with the conserved motif was thus indicated. These T cells were commonly present in patients with idiopathic SS and are strongly suggested to most likely be involved in the pathogenesis of both HTLV-I-associated and idiopathic SS. PMID- 10679128 TI - Correction PMID- 10679127 TI - High levels of IL-17 in rheumatoid arthritis patients: IL-15 triggers in vitro IL 17 production via cyclosporin A-sensitive mechanism. AB - Recent data suggest that IL-15 plays an important role in the pathogenesis of rheumatoid arthritis. In the present study, we hypothesized that elevated in the joints of rheumatoid arthritis, but not osteoarthritis, patients, IL-15 may exert its proinflammatory properties via the induction of IL-17, a cytokine known to stimulate synoviocytes to release several mediators of inflammation including IL 6, IL-8, GM-CSF and PGE2. To test this hypothesis, we first measured the levels of IL-17 and IL-15 using specific ELISA and found that synovial fluids of patients with rheumatoid arthritis, but not with osteoarthritis, contain high levels of these cytokines. A strong correlation between IL-15 and IL-17 levels in synovial fluids was observed. Among tested factors, LPS and TNF-alpha failed, IL 15 and IL-2 were equipotent, and PMA + ionomycin was far more efficient in the induction of IL-17 secretion by PBMCs isolated from healthy blood donors. Interestingly, synovial fluid cells, in contrast to PBMCs isolated from patients with rheumatoid arthritis, but not osteoarthritis, respond to PMA + ionomycin with much lower, comparable to IL-15-triggered IL-17 secretion. Moreover, PMA + ionomycin-triggered IL-17 secretion is completely or partially blocked in the presence of low doses of cyclosporin A or high doses of methylprednisolone, respectively. IL-15-triggered IL-17 secretion by PBMCs was completely inhibited by these drugs. Thus, our results suggest for the first time that IL-15 may represent a physiological trigger that via cyclosporin A and steroid sensitive pathways leads to the overproduction of IL-17 in the joints of rheumatoid arthritis patients. PMID- 10679130 TI - Effect of a comprehensive infection control program on the incidence of infections in long-term care facilities. AB - BACKGROUND: Control of infection within the long-term care facility is a daunting problem. Elderly patients are at high risk for contracting infection because of reduced innate immunity, malnutrition, and the presence of chronic medical conditions. This small study tested the effect of developing and implementing a comprehensive preventive infection control program in the long-term care setting and examined the resultant incidence of infections. METHODS: Eight private, freestanding, long-term care facilities in urban and suburban settings were selected for the study. The 4 test sites had a total of 443 beds; there were 447 beds in 4 matched control sites. Data on infection rates were accrued in both preintervention and intervention years. The control homes maintained their existing infection control policies and procedures. The test homes were provided with an infection control educational program and replaced all currently used germicidal products with single-branded products for a 12-month period. A criteria-based standardized infection control surveillance system was used to monitor and report infections in all facilities. RESULTS: In the preintervention year, the test sites experienced 743 infections (incidence density rate, 6.33) and the control homes experienced 614 infections (incidence density rate, 3.39). In the intervention year, the test homes reported 621 infections, a decrease of 122 infections (incidence density rate, 4.15); in the control homes, the number of infections increased slightly, to 626 (incidence density rate, 3.15). The greatest reduction in infections in the test homes was in upper respiratory infections (P =.06). CONCLUSIONS: This study provides additional evidence that a comprehensive infection control program that includes handwashing and environmental cleaning and disinfecting may help reduce infections among the elderly residing in long-term care settings. PMID- 10679131 TI - Microbiologic survey of long-term care facilities. AB - BACKGROUND: We undertook a microbiologic survey of long-term care facilities to categorize bacteria found in cultures of residents. Culture and sensitivity data were collected on 566 samples from indwelling bladder catheters, percutaneous gastrostomy tubes, nares, stool, wounds, pressure ulcers, and tracheostomies in 25 Nebraska and Iowa facilities. Information was also collected on resident factors (eg, presence of indwelling urinary catheter, prior antibiotic administration) and institutional variables (eg, number of beds, nosocomial infection rates). RESULTS: There were 478 gram-negative isolates, the leading organisms being Pseudomonas aeruginosa, Proteus mirabilis, Escherichia coli, and Klebsiella pneumoniae. There were 221 gram-positive isolates, the most frequently seen of which were enterococci and Staphylococcus aureus. Of the 442 residents sampled in the study, 168 (38%) were taking, or had within the previous month been taking, a systemic antibiotic. Quinolones were the most frequently prescribed antibiotic class. The institutional prevalence of urinary catheterization averaged 6.7%. CONCLUSIONS: Significant antibiotic pressure exists in long-term care facilities, a fact that is reflected in antibiotic resistance patterns. A variety of gram-positive and gram-negative bacteria were found in nursing home culture specimens. PMID- 10679132 TI - Surgical site infections at the National Cancer Institute in Mexico: a case control study. AB - OBJECTIVES: To quantify the surgical infection rate and to identify risk factors associated with surgical site infection. METHODS: We conducted a case-control study of all surgical patients between January 1, 1993, and June 30, 1994. The frequency of surgical site infection per 100 surgeries was calculated. The odds ratio (OR) was estimated by using logistic regression analysis. SETTING: A 130 bed tertiary-care teaching hospital for adult patients with cancer. RESULTS: The study followed 3372 surgeries. Three hundred thirteen patients had a surgical site infection (rate per 100 surgeries: 9. 30). The risk factors associated with surgical site infection were diabetes mellitus (OR = 2.5, 95% confidence interval [CI] = 1.27-4. 91), obesity (OR = 1.76, 95% CI = 1.14-2.7), presence of surgical drains for >5 and <16 days (OR = 1.84, 95% CI = 1.02-3.31), and presence of surgical drains for >/=16 days (OR = 2.14, 95% CI = 1. 0-4.6). The bacteria most frequently isolated were Escherichia coli 38 (21.8% of the total of microorganisms found), Pseudomonas sp 22 (12.6%), Staphylococcus aureus 16 (9.2%), and coagulase-negative Staphylococcus 25 (13.6%). The coexistence of other nosocomial infections was greater among the cases (OR = 1.8, 95% CI = 1.1 3.1) than in the control group. CONCLUSIONS: The surgical site infection rate in our hospital is slightly higher than the rates reported for general hospitals. The risk factors associated with surgical site infection are similar to those previously reported. Diabetes mellitus, obesity, and prolonged presence of a surgical drain increased the risk of infection. PMID- 10679133 TI - Survey on attitudes toward HIV-infected individuals and infection control practices among dentists in Mexico City. AB - BACKGROUND: The teaching of infection control is gradually being introduced at dental schools in Mexico. However, most practicing dentists have limited access to current infection control standards. Deficiencies of knowledge with regard to blood-borne pathogens such as HIV and hepatitis B virus may influence attitudes toward infected individuals and reduce compliance with infection control recommendations. OBJECTIVE: The purpose of this study was to assess (1) attitudes toward HIV-infected patients and hepatitis B virus-infected patients and (2) infection control knowledge and practices among dental practitioners in Mexico City. METHOD: A total of 196 dentists were interviewed by means of a questionnaire with Likert-type scales and open-ended questions (response rate, 86.1%). RESULTS: Most respondents had no previous social or professional contact with HIV-positive individuals. Nine percent indicated that they had knowingly treated HIV-positive patients. Perceived professional and moral obligations to treat HIV-positive patients were high. Thirty-five percent of the respondents perceived the risk of HIV infection as "considerable" to "very strong." The risk of hepatitis B infection was considered significantly higher than the risk of HIV infection (P <.01); however, 78% of the respondents had not been immunized against hepatitis B. Reported use of personal protective equipment was high. Most respondents used dry heat sterilization. The principal disinfectants used were quaternary ammonium compounds, bleach, and glutaraldehyde. Fifty-four percent of the respondents acknowledged that clinical precautions reduced occupational risks. CONCLUSIONS: This survey revealed contradictory attitudes toward HIV positive individuals and limited understanding of infection control recommendations. Educational and regulatory efforts are needed to promote better adherence to current infection control standards. PMID- 10679134 TI - Watching the bottom line: enhancing the role and impact of infection control in a managed care environment. AB - Health care expenditures exceeded $1 trillion in 1997, with projections for $2.2 trillion in expenditures by 2008. Decreasing organizational revenues and efforts to reduce operating costs have had a direct impact on infection control programs. Senior managers in hospitals and other provider organizations are focusing on achieving and maintaining revenues while controlling costs. Infection control professionals must align themselves and their programs with these organizational goals by (1) identifying areas in which the infection control program can support and enhances revenues, (2) facilitating the avoidance of excess costs for care, especially those related to nosocomial infection, (3) identifying opportunities for cost reduction through value analysis, and (4) participating in efforts to measure and prevent other adverse outcomes of care. PMID- 10679135 TI - The fifth evolutionary era in infection control: interventional epidemiology. AB - A historical review of infection control over the last 4 decades indicates that the field has evolved from being one whose investigative work laid the foundation for understanding the chain of infection to an influential profession whose research on effective prevention methods have revoluntionized clinical practice throughout the world. Underlying our successes is the fact that growth in the profession has brought with it an enormous expansion in responsibilities, which in turn has impacted, in some cases severely, the personnel and time resources of infection control departments. At the same time, the economic pressures brought on by the upheavals in the business of health care have trickled down wherein it now influences the makeup and effectiveness of infection control programs. To continue with our mission of reducing morbidity and mortality, and perhaps to avoid a diminishing of our own professional influence, it will become essential that new approaches to the management of infection control programs be implemented. The approach must start by incorporating a basic mandate for change in the infection control professional. PMID- 10679136 TI - Reengineering and infection control programs: commentary and a case study. AB - Transformation of the health care system has been an ongoing process for generations, but many changes in the past 2 decades have focused on reducing costs in concert with rapidly changing technologies and demands for high quality care. Many cost-containment efforts in the 1990s are characterized by attempts to apply the business model for "reengineering the corporation" to health care systems. This commentary reviews principles of reengineering and how strategies to reduce costs through market forces, competition, and downsizing can result in substantial problems for bureaucratic organizations unaccustomed to rapid change and innovation. A case study drawn from experiences of a large metropolitan academic health care system is presented, with specific focus on lessons that will be helpful to infection control professionals (ICPs) confronted with similar situations. PMID- 10679137 TI - Effect of a comprehensive program to reduce infectious waste. PMID- 10679138 TI - Direct costs associated with a nosocomial outbreak of Salmonella infection: an ounce of prevention is worth a pound of cure. AB - BACKGROUND: Nosocomial outbreaks of Salmonella infections in Australia are an infrequent but significant source of morbidity and mortality. Such an outbreak results in direct, measurable expenses for acute care management, as well as numerous indirect (and less quantifiable) costs to those affected, the hospital, and the wider community. This article describes the significant direct costs incurred as a result of a nosocomial outbreak of Salmonella infection involving patients and staff. METHOD: Information on costs incurred by the hospital was gathered from a number of sources. The data were grouped into 4 sections (medical costs, investigative costs, lost productivity costs, and miscellaneous) with use of an existing tool for calculating the economic impact of foodborne illness. RESULTS: The outbreak cost the hospital more than AU $120, 000. (US $95,000). This amount is independent of more substantial indirect costs. CONCLUSION: Salmonella infections are preventable. Measures to aid the prevention of costly outbreaks of nosocomial salmonellosis, although available, require an investment of both time and money. We suggest that dedication of limited resources toward such preventive strategies as education is a practical and cost-effective option for health care facilities. PMID- 10679139 TI - Expanding the role of the infection control professional in the cost-effective use of antibiotics. AB - There is a growing demand that health care expenses be contained and that excessive and inappropriate use of antibiotics be eliminated. At the University of California, San Diego Medical Center, strategies aimed at controlling drug usage and subsequently reducing costs have been implemented and found to be effective. Mechanisms designed to achieve such goals without diminishing quality of care involve expanding the role of the infection control professional (ICP) while implementing antibiotic control stratagems such as antimicrobial utilization teams, antibiotic order sheets, audits of use, automatic stop orders, computer-assisted management, drug use reviews, educational efforts, formulary practice, restricted drug policies, and target drug monitoring. The infection control professional, as well as other members of the antimicrobial utilization team, contributes to the promotion of the appropriate use of antibiotics in part by identifying individual cases in which antibiotics might be used inappropriately, such as for the treatment of colonization rather than infection or when appropriate microbiologic testing has not been carried out. PMID- 10679140 TI - "Where's the science"? PMID- 10679141 TI - Clinical and economic consequences of nosocomial catheter-related bacteriuria. AB - Indwelling catheters are strongly associated with the development of bacteriuria, which can lead to significant morbidity in hospitalized patients. This report, a review of the literature, evaluates the infectious outcomes of patients with indwelling catheters to determine the precise clinical and economic impact of catheter-related infection. Statistical pooling was used to estimate the incidence of bacteriuria in hospitalized patients with indwelling catheters. In addition, the proportion of patients with catheter-related bacteriuria in whom symptomatic urinary tract infection and bacteremia will develop was estimated through quantitative synthesis of previous reports. Costs were estimated by using microcosting techniques. Of patients who have indwelling catheters for 2 to 10 days, bacteriuria is expected to develop in 26% (95% confidence interval [CI], 23% to 29%). Among patients with bacteriuria symptoms of urinary tract infection will develop in 24%, (95% CI, 16% to 32%), and bacteremia from a urinary tract source will develop in 3.6% (95% CI, 3.4% to 3.8%). Each episode of symptomatic urinary tract infection is expected to cost an additional $676, and catheter related bacteremia is likely to cost at least $2836. Given the clinical and economic burden of urinary catheter-related infection, infection control professionals and hospital epidemiologists should use the latest infection control principles and technology to reduce this common complication. PMID- 10679142 TI - Gastrointestinal hormones. AB - Solomon A. Berson, M.D., the first Murray M. Rosenberg Professor and Chair of the Department of Medicine at Mount Sinai from 1968 until his death in 1972, and Rosalyn S. Yalow, Ph.D., 1977 Nobel Laureate in Medicine or Physiology and Solomon A. Berson Distinguished Professor-at-Large, brought meticulous quantitation and new vistas to all of clinical medicine and biomedical science through the application of their technique of radioimmunoassay. I was fortunate to know and work with them for many years. In 1972, while I was an NIH Fellow in gastroenterology at Mount Sinai, Dr. Berson suggested that I pursue my research in their laboratory at the Bronx Veterans Administration Hospital. Dr. Berson died one month after I began my research in the Bronx. Yalow and Berson had already discovered big gastrin (G-34), but much work with gastrin remained to be done. Challenging work with secretin, cholecystokinin, and a host of other gut peptides, would keep the Mount Sinai group at the forefront of this exciting field. PMID- 10679144 TI - Robert Jackson Williams (1931-1999). PMID- 10679145 TI - Cryopreservation of mouse oocytes using a medium with low sodium content: effect of plunge temperature. AB - The effect of various combinations of plunge temperature and thawing protocol on the survival and viability of mouse oocytes was examined. The oocytes were frozen either in a standard freezing medium (ETFM, embryo transfer freezing medium) or in a low-sodium, choline-based freezing medium (CJ2), with 1.5 M 1,2-propanediol and 0.1 M sucrose, and using a conventional slow cooling method. The criteria used to assess survival were morphological state after thawing (intact or lysed), ability to become fertilized, and ability to develop to the two-cell, morula, and blastocyst stage in vitro. Oocytes frozen in CJ2 and plunged into liquid nitrogen (LN(2)) from -10, -20, or -33 degrees C remained intact and developed to the blastocyst stage at significantly higher rates than oocytes frozen in ETFM. For oocytes plunged into LN(2) from -33 degrees C, very rapid thawing (10 s in 30 degrees C water) was more detrimental than rapid or slow thawing (holding in air at room temperature for 10 or 30 s, respectively, prior to submersion in water at 30 degrees C for 10 s). By contrast, oocytes plunged into LN(2) from -10 or -20 degrees C survived better when thawing was very rapid or rapid than when thawing was slow. With the current protocol CJ2 was very effective over a wide range of plunge temperatures (-20 to -33 degrees C), although the optimal thawing protocol depended on the particular plunge temperature. Over 90% of oocytes surviving after slow cooling in CJ2 to -33 degrees C could be plunged to -196 degrees C with little or no further damage. PMID- 10679146 TI - Rapid MR imaging of cryoprotectant permeation in an engineered dermal replacement. AB - Magnetic resonance (MR) imaging is a powerful technique for monitoring the permeation of cryoprotective agents (CPAs) inside tissues. However, the techniques published until now suffer from inherently long imaging times, limiting the application of these techniques to slow diffusion processes and large CPA concentrations. In this study, we present a rapid MR imaging technique based on a CHESS-FLASH scheme combined with Keyhole image acquisition. This technique can image the fast permeation of Me(2)SO solutions into freeze-dried artificial dermal replacements for concentrations down to 10% v/v. Special attention is given to evaluating the technique for quantitative analysis. PMID- 10679147 TI - Effect of phospholipase A2 inhibitors on the release of arachidonic acid and cell viability in cold-stored hepatocytes. AB - We investigated the effect of phospholipase A(2) (PLA(2)) inhibitors on PLA(2) activity and cell viability in cold-stored rat hepatocytes. The cells were radiolabeled with [(3)H] arachidonic acid (AA) and cold stored in the University of Wisconsin (UW) solution containing various PLA(2) inhibitors. PLA(2) activity was determined by measuring the total free (cellular + supernatant) AA by thin layer chromatography after inhibiting reacylation of free AA with inhibitors of energy production (carbonyl cyanide m-chlorophenylhydrazone + iodoacetate). Aristolochic acid, chlorpromazine, and quinacrine in the UW solution showed a significant inhibitory effect throughout 48 h cold storage but only at relatively high concentration. PLA(2) activity was also suppressed (58% of control) by trifluoperazine (50 microM), but its effect was limited to only 24 h. In contrast, pretreatment of the cells prior to hypothermic preservation with trifluoperazine (10 to 100 microM) suppressed PLA(2) activity during 48 h storage. Inclusion of calmodulin antagonist W-7 did not affect PLA(2) activity. Thus, the inhibitory activity of these agents appears unrelated to Ca-calmodulin phospholipid interaction but to have an inhibitory effect on PLA(2) activity. To study the effects of PLA(2) inhibitors on cell viability, lactate dehydrogenase (LDH) release was measured in the presence or absence of inhibitors upon rewarming cold-stored cells in Krebs-Henseleit buffer for 2 h at 37 degrees C. None of the inhibitors tested improved cell viability after 48 h storage. Thus, although PLA(2) inhibitors blocked PLA(2) activity, there was no suppression of LDH release. PLA(2) may play a minor role in preservation/reperfusion injury to cold-stored hepatocytes. PMID- 10679148 TI - Fertilizing potential of mouse spermatozoa cryopreserved in a medium containing whole eggs. AB - Experiments were conducted to improve survival of mouse spermatozoa through the cryopreservation process. In the first experiment, percentages of motile spermatozoa and fertilizing capacities of spermatozoa were evaluated when mouse spermatozoa were cryopreserved using three previously reported cryopreservation media: (1) 18% raffinose in 3% skim milk; (2) Tes/Tris medium containing 25% egg yolk and 1.25% glycerol; and (3) PBS containing 18% raffinose and 1.75% glycerol, each at three different cooling rates (-3, -10, and -50 degrees C/min). Spermatozoa frozen in the skim milk/raffinose medium exhibited the highest percentage of motile spermatozoa (39%) when cells were frozen at -10 degrees C/min (P<0.05). The second experiment evaluated the effects of modifying the Tes/Tris/egg yolk medium, comparing different concentrations of egg yolk, BSA, and sodium dodecyl sulfate. Reducing egg yolk from 25% of the medium volume to 5%, increased percentages of motile spermatozoa after cryopreservation from 29 to 36% (P<0.05). Addition of 1% BSA and sodium dodecyl sulfate to medium containing 5% egg yolk further improved percentages of motile spermatozoa after freezing. In the final experiment, 20% whole egg was substituted for 5% egg yolk and 1% BSA used in previous experiments and resulted in percentages of motile spermatozoa (51%) equal to that of the skim milk-raffinose medium. However, fertility rates were higher (68%) than for spermatozoa frozen in the skim milk-raffinose medium (P < 0.05) and were comparable to the fertility rates of fresh spermatozoa (77%; P>0.05). In conclusion, freezing mouse spermatozoa in a medium containing 20% whole egg, 0.035% sodium dodecyl sulfate, and 1.25% glycerol using a cooling rate of -10 degrees C/min preserves the motility and fertilization capacity of mouse spermatozoa. PMID- 10679149 TI - Cryopreservation of seabream (Sparus aurata) spermatozoa. AB - The aim of this research was to optimize protocols for freezing spermatozoa of seabream (Sparus aurata). All the phases of the cryopreservation procedure (sampling, choosing the cryoprotective extender, cooling, freezing, and thawing) were studied in relation to the species of spermatozoa under examination, so as to be able to restore on thawing the morphological and physiological characteristics of fresh semen. Seabream spermatozoa were collected by stripping and transported to the laboratory chilled (0-2 degrees C). Five cryoprotectants, dimethyl sulfoxide (Me(2)SO), ethylene glycol (EG), 1,2-propylene glycol (PG), glycerol, and methanol, were tested at concentrations between 5 and 15% by volume to evaluate their effect on the motility of semen exposed for up to 30 min at 26 degrees C. The less toxic cryoprotectants, 10% EG, 10% PG, and 5% Me(2)SO, respectively, were added to 1% NaCl to formulate the extenders for freezing. The semen was diluted 1:6 with the extender, inserted into 0.25-ml plastic straws by Pasteur pipette, and frozen using a cooling rate of either 10 or 15 degrees C/min to -150 degrees C followed by transfer and storage in liquid nitrogen (-196 degrees C). The straws were thawed at 15 degrees C/s. On thawing, the best motility was obtained with 5% Me(2)SO, although both 10% PG and EG showed good results; no differences were found between the two freezing gradients, although semen frozen with the 10 degrees C/min gradient showed a slightly higher and more prolonged motility. PMID- 10679150 TI - Cell-cell contact affects membrane integrity after intracellular freezing. AB - The response of cells to freezing depends critically on the presence of an intact cell membrane. During rapid cooling, the cell plasma membrane may no longer be an effective barrier to ice propagation and can be breached by extracellular ice resulting in the nucleation of the supercooled cytoplasm. In tissues, the formation of intracellular ice is compounded by the presence of cell-cell and cell-surface interactions. Three different hamster fibroblast model systems were used to simulate structures found in organized tissues. Samples were supercooled to an experimental temperature on a cryostage and ice nucleated at the constant temperature. A dual fluorescent staining technique was used for the quantitative assessment of the integrity of the cell plasma membrane. A novel technique using the fluorescent stain SYTO was used for the detection of intracellular ice formation (IIF) in cell monolayers. The cumulative incidence of cells with a loss of membrane integrity and the cumulative incidence of IIF were determined as a function of temperature. Cells in suspension and individual attached cells showed no significant difference in the number of cells that formed intracellular ice and those that lost membrane integrity. For cells in a monolayer, with cell-cell contact, intracellular ice formation did not result in the immediate disruption of the plasma membrane in the majority of cells. This introduces the potential for minimizing damage due to IIF and for developing strategies for the cryoprotection of tissues during rapid cooling. PMID- 10679151 TI - A two-parameter model of cell membrane permeability for multisolute systems. AB - A two-parameter model of cell osmotic response (F. W. Kleinhans, 1998, Cryobiology 37, 271-289) is expanded for multisolute systems. The cell water volume W and intracellular osmolalities of N solutes are related as W[1 + L(p)RTSigma(N)(i=1)(M(i)/P(i))] = W(0)[1 + L(p)RTSigma(N)(i=1)(M(0)i)/P(i))], where M(i) is the intracellular osmolality of the ith solute (i = 1 ellipsis N), P(i) is the membrane permeability of the ith solute, L(p) is the membrane hydraulic conductivity, R is the gas constant, T is the absolute temperature, and the subscript "0" denotes the initial values at time zero. The above formula allows calculating the final (equilibrium) volume when all entities are permeable. Simple algebraic expressions for calculation of the number and magnitude of transient maximum volume excursions are presented. These simple expressions can all be calculated by hand on a pocket calculator. Practical examples of one-, two-, and three-solute systems are discussed. Special attention has been given to situations when systems contain an impermeable component. All formulas are simple to use for optimization of variety of cryobiological protocols. Application of the theory for optimization of addition and dilution of a permeable cryoprotectant is also discussed. PMID- 10679152 TI - Transplantation of mammalian livers following freezing: vascular damage and functional recovery. AB - We transplanted rat livers cryopreserved at high subzero temperatures with a protocol that mimics freezing in freeze-tolerant animals. The results of nine transplants show that: (a) every single transplanted liver produced bile, which suggests that the cryopreserved livers retained some physiological function; (b) eight of the animals survived between 2 and 4 h with loss of microvascular integrity which suggests that transplantation failure is related to the circulation and tests of bile production are not indicative of transplantation success; and (c) one animal survived for 5 days with an intact circulation which might be due to an unidentified technical variation or could indicate that when the circulation recovers animals with transplanted livers survive. PMID- 10679153 TI - Molecular phylogeny of north mediterranean freshwater barbs (genus Barbus: cyprinidae) inferred from cytochrome b sequences: biogeographic and systematic implications. AB - We investigated phylogenetic relationships among north Mediterranean species of the genus Barbus using sequences of the cytochrome b gene. Our results indicate that the species belong to two major clades that are consistent with those previously defined from morphological features. The first clade includes species ranging from France to the Black Sea. In this clade, there is a well-supported monophyletic group of large-sized fluvio-lacustrine barbs; however, the monophyly of the small-sized rheophilic species is not clear. The second clade comprises species found in Spain, Greece, and Asia Minor and probably represents the oldest group present in the north Mediterranean rivers. In general, there is good concordance between geography and phylogenetic relationships. These results are compared to those from previous morphological- and allozyme-based studies and demonstrate widespread discordance and polyphyly in the traditional taxonomy of the genus Barbus. This study is one of the first reporting the phylogenetic and biogeographic relationships of a genus that is widely distributed in European rivers and contains species that are a major component of the European ichthyofauna. PMID- 10679154 TI - An 18S rDNA-based molecular phylogeny of aphidiinae (Hymenoptera: braconidae). AB - We have obtained a molecular phylogeny of the subfamily Aphidiinae (Hymenoptera: Braconidae) by sequencing the 18S rDNA in 37 aphidiine taxa. Approximately 1857 nucleotides were sequenced in each species. Evolutionary relationships were established by comparing the results of maximum-parsimony, maximum-likelihood, and distance analyses. The most variable region of this gene, V4 (approx 403 nucleotides), was employed to establish the basality of the tribe Ephedrini within this subfamily. All phylogenetic reconstructions yielded trees with very similar topologies that confirmed the existence of two of the four traditionally accepted tribes, Ephedrini and Praini, but questioned the existence of Trioxini and Aphidiini. To better ascertain the status of some groups, the same analyses were repeated with a reduced taxonomic sample in which some species that produced systematic errors in the former phylogenetic reconstructions had been removed. The results from this second analysis favor either the three-tribes hypothesis (Ephedrini, Praini, and Aphidiini) or a new classification with at least five tribes (Ephedrini, Praini, Monoctonini, Trioxini, and Aphidiini). The 18S rDNA gene is a useful marker to recover relationships not only at the tribe but also at the subtribe and genus levels in this group. The natural status of some traditionally accepted clusters is also corroborated with the present data whereas the placement of other clusters is questioned or remains unresolved. PMID- 10679155 TI - Molecular phylogenetics of subfamily Calamoideae (Palmae) based on nrDNA ITS and cpDNA rps16 intron sequence data. AB - Phylogenetic relationships among the 22 genera of the palm subfamily Calamoideae were investigated using DNA sequence data from the nuclear ribosomal internal transcribed spacer (ITS) region and the chloroplast rps16 intron. The rps16 intron displayed low levels of variation, corroborating previous reports that the chloroplast genome of palms is highly conserved. High levels of within-individual polymorphism were identified in the ITS region, indicating that concerted evolution is not effectively homogenizing the ITS repeats. In the majority of cases, multiple clones from individuals resolved as monophyletic. However, the high levels of homoplasy in the ITS dataset, along with generally poor jackknife support for many clades, led to concerns that topologies obtained from these data might be unreliable. Nevertheless, congruence between trees based on ITS data alone and those based on rps16 intron data was high. Simultaneous analyses of both datasets yielded well-resolved topologies with high levels of jackknife support. A number of exciting groups emerged from the analyses: the African rattan clade comprising the endemic African rattan genera Laccosperma, Eremospatha, and Oncocalamus; the Lepidocaryeae-Raphia clade comprising the fan leaved New World tribe Lepidocaryeae and the African genus Raphia; and the Asian clade comprising all Asian genera except Eugeissona. The position of Eugeissona was variable, although it did not resolve inside any of the three major clades mentioned above. PMID- 10679156 TI - Molecular phylogenetics of Calamus (Palmae) and related rattan genera based on 5S nrDNA spacer sequence data. AB - Phylogenetic relationships among the rattan palm genera Calamus, Daemonorops, Ceratolobus, Calospatha, Pogonotium, and Retispatha were investigated using DNA sequences from the nontranscribed spacer of 5S nrDNA. Moderate levels of intragenome polymorphism were identified, indicating that concerted evolution is not completely homogenizing the multiple copies of the 5S nrDNA repeat present in the nuclear genome. The existence of intragenome polymorphism did not excessively interfere with phylogeny reconstruction because, in the majority of cases, multiple clones obtained from individual species were resolved as monophyletic groups. The highly speciose genus Calamus was found to be nonmonophyletic with all five remaining genera being embedded within it. A number of major lineages within Calamus were resolved, one of which included the monotypic genus Calospatha, another included the monotypic genus Retispatha, and a third included a monophyletic group comprising Daemonorops, Ceratolobus, and Pogonotium. While the findings indicate that generic circumscriptions require revision, a nomenclatural solution was not sought at this stage because inadequate sampling and lack of support at basal nodes suggested that the topologies obtained might not be entirely reliable. Under these circumstances, name changes to such an important group would be both unhelpful and irresponsible. PMID- 10679157 TI - Study of the evolutionary relationships among Limonium species (Plumbaginaceae) using nuclear and cytoplasmic molecular markers. AB - The genus Limonium, due to the patchiness of the natural habitats of its species as well as the high frequency of hybridization and polyploidy and the possibility of reproduction by apomixis, provides an example of all the principal mechanisms of rapid speciation of plants. As an initial study of evolution in this genus, we have analyzed intra- and interspecific variability in 17 species from section Limonium, the largest in the genus, based on RFLPs of cpDNA and nuclear rDNA ITS sequences. In the cpDNA analysis, 21 restriction enzymes were used, resulting in 779 fragments, 490 of which were variable and 339 parsimony informative. L. furfuraceum exhibited two relatively divergent cpDNA haplotypes. The relationships found among the species based on cpDNA restriction fragments were coincident using different methods of phylogenetic analysis. Due to the presumed reticulate evolution in the genus Limonium, the comparison of these results with data from the nuclear DNA was necessary; ITS sequences were analyzed. The final alignment contained 488 characters, of which 198 were variable and 156 parsimony informative. Two relatively divergent ITS types were present at the intraindividual level in L. delicatulum, a triploid species. Each type was related to ITS from different groups of diploid Limonium species, one with a base haploid chromosome number n = 8 (represented by L. cossonianum) and the other with n = 9 (represented by L. minutum). The different phylogenetic inference methods used for the analysis of ITS sequences rendered very similar topologies. In general, the relationships among the species studied were coincident with those obtained with the chloroplast genome. Both nuclear and cytoplasmic markers support the polyphyly of section Limonium, with at least two species, L. narbonense and L. vulgare, clearly divergent from the rest. Moreover, the remaining subsections into which section Limonium is currently divided seem to be artificial. PMID- 10679158 TI - A new molecular phylogenetic hypothesis for the evolution of freshwater eels. AB - Phylogenetic analysis of a segment of the mitochondrial 16S rDNA of eight Anguilla species from the Indo-Pacific region and from the North Atlantic revealed that the genus Anguilla appears to be surprisingly young, based upon the small observed maximum genetic distance of 4.8% and the high degree of morphological similarity among the species. The placement of A. marmorata as the most ancestral lineage suggests that the genus is likely to have originated in the Indo-Malayian region, from which it quickly spread. Two Pacific species, A. obscura and A. japonica, branched next. A. japonica was placed as sister group to all remaining species, which formed three clades: the first comprising A. australis, the second A. reinhardti and A. mossambica, and the third A. anguilla and A. rostrata. All analyzed specimens of A. rostrata originating from southern New Jersey to Nova Scotia had identical mitotypes, while five mitochondrial genotypes were found in Europe differing by zero to two substitutions. The two Atlantic eel species are very closely related; all surveyed specimens of A. anguilla differ by three to five substitutions from their American allies, corroborating the existence of two distinct biological species. This was also confirmed by restriction analysis of a 350-bp segment of the cytochrome b, in which American specimens were distinct in sharing a single diagnostic restriction site of HinfI. Our results suggest little to no gene flow between the two nominal Atlantic eel species. PMID- 10679159 TI - DNA archives and our nearest relative: the trichotomy problem revisited. AB - Ever since Thomas H. Huxley correctly identified the chimpanzee and the gorilla as the two closest relatives of the human, the problem of the relationship among the three species ("the trichotomy problem") has remained unresolved. Comparative morphology and other classical methods of biological investigation have failed to answer definitively whether the chimpanzee or the gorilla is the closest relative of the human species. DNA sequences, both mitochondrial and nuclear, too, have provided equivocal solutions, depending on the region of the genome analyzed. Random sorting of ancestral allelic lineages, sequence convergence, and sequence exchanges between alleles or duplicated loci have been identified as likely factors confounding the interpretation of the interrelationships among the three species. In the present study most of these difficulties are overcome by identifying evolutionary causes that might potentially provide misleading information. Altogether, 45 loci consisting of 46, 855 bp are analyzed. About 60% of the loci and approximately the same proportion of phylogenetically informative sites support the human-chimpanzee clade. The remaining 40% of loci and sites support the two alternatives equally. It is demonstrated that, while incompatibility between loci can be explained by random sorting of allelic lineages, incompatibility within loci must be attributed largely to the joint effect of recombination and genetic drift. The trichotomy problem can be properly addressed only within this framework. PMID- 10679160 TI - Ancestral lineages of arbuscular mycorrhizal fungi (Glomales). AB - Using new and existing 18S rRNA sequence data, we show that at least five species of glomalean fungi lie outside the previously defined families and diverged very early in the evolution of that group. These five fungi would have been missed by many previous ecological studies because their sequences are not well matched to available taxon-specific primers and they do not stain well with the standard reagents used for morphological analysis. Based upon spore morphology, these species are currently assigned to Glomus and Acaulospora, and two of the species are dimorphic, exhibiting spore stages of both genera. This suggests that dimorphic spores are the ancestral state for the order and that one or the other morphology was lost in various lineages. Our analyses also show that Geosiphon pyriforme, a symbiont with cyanobacteria, is not necessarily a sister group of the Glomales; instead, it may be derived from mycorrhizal ancestors. PMID- 10679161 TI - The complete external transcribed spacer of 18S-26S rDNA: amplification and phylogenetic utility at low taxonomic levels in asteraceae and closely allied families. AB - For molecular phylogenetic reconstruction of some intrageneric groups of plants, a DNA region is needed that evolves more rapidly than the internal transcribed spacer (ITS) of the 18S-26S nuclear ribosomal DNA (nrDNA) repeat. If the region identified is nuclear, it would also be desirable for it to undergo rapid concerted evolution to eliminate problems with coalescence. The external transcribed spacer (ETS) of the nrDNA repeat has shown promise for intrageneric phylogenetic reconstruction, but only the 3' end of the region has been utilized for phylogenetic reconstruction and "universal" primers for PCR amplification have been elusive. We present a method for reliably amplifying and sequencing the entire ETS throughout Asteraceae and some closely allied families. We also show that the ETS is more variable and phylogenetically informative than the ITS in three disparate genera of Asteraceae-Argyranthemum (tribe Anthemideae), Asteriscus (tribe Inuleae), and Helianthus (tribe Heliantheae). The full ETS was amplified using a primer (ETS1f) within the intergenic spacer in combination with a primer (18S-2L) in the 5' end of the highly conserved 18S gene. ETS1f was designed to correspond to a highly conserved region found in Helianthus and Crepis, which are in separate subfamilies of Asteraceae. ETS1f/18S-2L primed in all of the tribes of Asteraceae as well as exemplar taxa from Campanulaceae, Goodeniaceae, and Calyceraceae. For both Argyranthemum and Asteriscus, we were able to directly sequence the ETS PCR products when a single band was produced. When multiple bands were produced, we gel-purified and occasionally cloned the band of interest before sequencing. Although PCR produced single bands for Helianthus species, it was necessary to clone Helianthus amplifications prior to sequencing due to multiple intragenomic ETS repeat types. Alignment of ETS sequences for Argyranthemum and Asteriscus was straightforward and unambiguous despite some subrepeat structure in the 5' end. For Helianthus, different numbers of large tandem subrepeats in different species required analysis of the orthology of the subrepeats prior to alignment. In all three genera, the ETS provided more informative variation for phylogenetic reconstruction and allowed better resolution of relationships than the ITS. Although cloned sequences from Helianthus differed, intragenomic clones consistently formed clades. This result indicated that concerted evolution was proceeding rapidly enough in ETS that species-specific phylogenetic signal was retained. It should be now be possible to use the entire ETS for phylogenetic reconstruction of recently diverged lineages in Asteraceae and at least three other families (approximately 26,000 species or about 8% of all angiosperms). PMID- 10679162 TI - The evolutionary history of the genus Timarcha (Coleoptera, Chrysomelidae) inferred from mitochondrial COII gene and partial 16S rDNA sequences. AB - The apterous genus Timarcha consists of three subgenera and more than 100 species in its Palearctic distribution, with specialized feeding on few plant families. Fifty-four sequences sampled from 31 taxa of the genus plus three outgroup leaf beetles were studied for their complete cytochrome oxidase II (COII) and a fragment of 16S rDNA mitochondrial genes, representing a total of about 1200 bp. Phylogenetic analyses using maximum-parsimony and distance methods for each gene separately and for the combined data set gave compatible topologies. The subgenus Metallotimarcha consistently appears in a basal position and is well differentiated from the remaining Timarcha, but no clear monophyletic grouping of Timarchostoma and Timarcha s. str. subgenera can be deduced from our analysis. Calibration of the molecular clock has been done using the opening of the Gibraltar Strait after the Messinian salinity crisis (about 5.5 MYA) as the biogeographic event causing disjunction of two particular taxa. Accordingly, the COII evolutionary rate has been estimated to be of 0.76 x 10(-8) substitution/site/year in Timarcha. Relation between phylogeny and host-plant use indicates widening of trophic regime as a derived character in Timarcha. PMID- 10679163 TI - Characterization of novel sequences from distantly related taxa by walking PCR. PMID- 10679182 TI - Regional differences in the CBF and BOLD responses to hypercapnia: a combined PET and fMRI study. AB - Previous fMRI studies of the cerebrovascular response to hypercapnia have shown signal change in cerebral gray matter, but not in white matter. Therefore, the objective of the present study was to compare (15)O PET and T *(2)-weighted MRI during a hypercapnic challenge. The measurements were performed under similar conditions of hypercapnia, which were induced by inhalation of 5 or 7% CO(2). The baseline rCBF values were 65.1 ml hg(-1) min(-1) for temporal gray matter and 28.7 ml hg(-1) min(-1) for white matter. By linear regression, the increases in rCBF during hypercapnia were 23.0 and 7. 2 ml hg(-1) min(-1) kPa(-1) for gray and white matter. The signal changes were 6.9 and 1.9% for the FLASH sequence and were 3.8 and 1. 7% for the EPI sequence at comparable echo times. The regional differences in percentage signal change were significantly reduced when normalized by regional flow values. A deconvolution analysis is introduced to model the relation between fMRI signal and end-expiratory CO(2) level. Temporal parameters, such as mean transit time, were derived from this analysis and suggested a slower response in white matter than in gray matter regions. It was concluded that the differences in the magnitude of the fMRI response can largely be attributed to differences in flow and that there is a considerable difference in the time course of the response between gray and white matter. PMID- 10679183 TI - Electrophysiological correlates of recollecting faces of known and unknown individuals. AB - We recorded brain potentials from healthy human subjects during a recognition test in order to monitor neural processing associated with face recollection. Subjects first attempted to memorize 40 faces; half were accompanied by a voice simulating that person speaking (e.g., "I'm Jimmy and I was a roadie for the Grateful Dead") and half were presented in silence. In the test phase, subjects attempted to discriminate both types of old faces (i.e., "named" and "unnamed" faces) from new faces. Recognition averaged 87% correct for named faces, 74% correct for unnamed faces, and 91% correct for new faces. Potentials to old faces were more positive than those to new faces from 300 to 600 ms after face onset. For named faces, the old-new ERP difference was observed at anterior and posterior scalp locations. For unnamed faces, the old-new ERP difference was observed only at posterior scalp locations. Results from a prior experiment suggest that these effects do not reflect perceptual priming of faces. The posterior portion of the old-new ERP difference was thus interpreted as a neural correlate of retrieval of visual face information and the anterior portion as an indication of retrieval of person-specific semantic information. PMID- 10679184 TI - Cortical activity related to accuracy of letter recognition. AB - Previous imaging and neurophysiological studies have suggested that the posterior inferior temporal region participates in tasks requiring the recognition of objects, including faces, words, and letters; however, the relationship between accuracy of recognition and activity in that region has not been systematically investigated. In this study, positron emission tomography was used to estimate glucose metabolism in 60 normal adults performing a computer-generated letter recognition task. Both a region of interest and a voxel-based method of analysis, with subject state and trait variables statistically controlled, found task accuracy to be: (1) negatively related to metabolism in the left ventrolateral inferior temporal occipital cortex (Brodmann's area 37, or ventrolateral BA 37) and (2) positively related to metabolism in a region of the right ventrolateral frontal cortex (Brodmann's areas 47 and 11, or right BA 47/11). Left ventrolateral BA 37 was significantly related both to hits and to false alarms, whereas the right BA 47/11 finding was related only to false alarms. The results were taken as supporting an automaticity mechanism for left ventrolateral BA 37, whereby task accuracy was associated with automatic letter recognition and in turn to reduced metabolism in this extrastriate area. The right BA 47/11 finding was interpreted as reflecting a separate component of task accuracy, associated with selectivity of attention broadly and with inhibition of erroneous responding in particular. The findings are interpreted as supporting the need for control of variance due to subject and task variables, not only in correlational but also in subtraction designs. PMID- 10679185 TI - The functional neuroanatomy and long-term reproducibility of brain activation associated with a simple finger tapping task in older healthy volunteers: a serial PET study. AB - We examined long-term reproducibility of the functional organization of the brain associated with a simple finger tapping movement using positron emission tomography (PET). Repeat measurements of regional cerebral blood flow were obtained in 10 individuals, ages 35 to 82 years (mean 52 years), at scanning sessions separated by 6 months. Although the functional neuroanatomy of hand movements has previously been investigated with PET by a number of groups, none has reported systematic investigation of the consistency of brain activation over an extended time. As expected, we found significant activation in the left precentral gyrus [Talairach coordinate (-32, -34, 52)], postcentral gyrus (-22, 48, 56), and supplementary motor area (SMA) (-2, -18, 52) at the initial study, consistent with previous studies in younger subjects. For the follow-up study we also found significant activation in the left precentral (-36, -28, 52) and postcentral (-28, -36, 52) gyri and in the SMA (2, -16, 56). Our group results demonstrate consistent anatomical location and extent of motor activation over time. More importantly, analysis of individuals confirmed the presence of consistent sites of activation in primary sensorimotor cortex and SMA over the 6 month interval in most subjects. A high degree of consistency in location of activation in the group, and within individuals, over time suggests that changes in loci of activation may be confidently monitored using the PET method. Evidence of individual differences in extent of activation over time highlights the need for caution when interpreting similar changes in patient studies. PMID- 10679186 TI - Distinguishing the functional roles of multiple regions in distributed neural systems for visual working memory. AB - We have investigated the human neural systems for visual working memory using functional magnetic resonance imaging to distinguish sustained activity during memory delays from transient responses related to perceptual and motor operations. These studies have identified six distinct frontal regions that demonstrate sustained activity during memory delays. These regions could be distinguished from brain regions in extrastriate cortex that participate more in perception and from brain regions in medial and lateral frontal cortex that participate more in motor control. Moreover, the working memory regions could be distinguished from each other based on the relative strength of their participation in spatial and face working memory and on the relative strength of sustained activity during memory delays versus transient activity related to stimulus presentation. These results demonstrate that visual working memory performance involves the concerted activity of multiple regions in a widely distributed system. Distinctions between functions, such as perception versus memory maintenance, or spatial versus face working memory, are a matter of the degree of participation of different regions, not the discrete parcellation of different functions to different modules. PMID- 10679187 TI - A PET investigation of the attribution of intentions with a nonverbal task. AB - Several authors have demonstrated that theory of mind is associated with a cerebral pattern of activity involving the medial prefrontal cortex. This study was designed to determine the cerebral regions activated during attribution of intention to others, a task which requires theory-of-mind skills. Eight healthy subjects performed three nonverbal tasks using comic strips while PET scanning was performed. One condition required subjects to attribute intentions to the characters of the comic strips. The other two conditions involved only physical logic and knowledge about objects' properties: one condition involved characters, whereas the other only represented objects. The comparison of the attribution of intention condition with the physical logic with characters condition was associated with rCBF increases in the right middle and medial prefrontal cortex including Brodmann's area (BA) 9, the right inferior prefrontal cortex (BA 47), the right inferior temporal gyrus (BA 20), the left superior temporal gyrus (BA 38), the left cerebellum, the bilateral anterior cingulate, and the middle temporal gyri (BA 21). The comparison of the physical logic with characters condition and the physical logic without characters condition showed the activation of the lingual gyri (BA 17, 18, 19), the fusiform gyri (BA 37), the middle (BA 21) and superior (BA 22, 38) temporal gyri on both sides, and the posterior cingulate. These data suggest that attribution of intentions to others is associated with a complex cerebral activity involving the right medial prefrontal cortex when a nonverbal task is used. The laterality of this function is discussed. PMID- 10679188 TI - Regulation of beta-catenin signaling in the Wnt pathway. AB - beta-Catenin not only regulates cell to cell adhesion as a protein interacting with cadherin, but also functions as a component of the Wnt signaling pathway. The Wnt signaling pathway is conserved in various organisms from worms to mammals, and plays important roles in development, cellular proliferation, and differentiation. Wnt stabilizes cytoplasmic beta-catenin and then beta-catenin is translocated into the nucleus where it stimulates the expression of genes including c-myc, c-jun, fra-1, and cyclin D1. The amounts and functions of beta catenin are regulated in both the cytoplasm and nucleus. Its molecular mechanisms are becoming increasingly well understood. PMID- 10679189 TI - Evidence for the carboxyl methylation of nuclear lamin-B in the pancreatic beta cell. AB - Lamins are intermediate filament proteins that constitute the main components of the lamina underlying the inner-nuclear membrane and serve to organize chromatin. Lamins (e.g., lamin-B) undergo posttranslational modifications (e.g., isoprenylation and methylation) at their C-terminal cysteine. Such modifications are thought to render optimal association of lamins with the nuclear envelop. Herein, we examined whether nuclear lamin-B undergoes carboxyl methylation in islet beta cells. A 65- to 70-kDa protein was carboxyl methylated in intact rat islets and clonal beta (HIT or INS) cells or in homogenates which could be immunoprecipitated using lamin-B antiserum. Incubation of purified HIT cell nuclear fraction with [(3)H]S-adenosyl methionine yielded a single carboxyl methylated protein peak (ca. 65-70 kDa); this protein was immunologically identified as lamin-B. Several methylation inhibitors, including acetyl farnesyl cysteine, a competitive inhibitor of protein prenyl cysteine methylation, inhibited the carboxyl methylation of lamin-B, indicating that the carboxyl methylated amino acid is cysteine. These findings, together with our recent observations demonstrating that inhibition of protein isoprenylation causes apoptotic death of the pancreatic beta cell, raise an interesting possibility that inhibition of C-terminal cysteine modifications of lamin-B might result in disruption of nuclear assembly, leading to further propagation of apoptotic signals, including DNA fragmentation and chromatin condensation. PMID- 10679190 TI - Evidence for translational repression of the SOCS-1 major open reading frame by an upstream open reading frame. AB - The suppressor of cytokine signalling 1 protein (SOCS-1) belongs to a novel family of cytokine inducible factors which function as inhibitors of the JAK/STAT pathway. While SOCS-1 previously has been described as a single-exon gene, here we present evidence for an additional 5' exon, separated by a 509 bp intron from exon 2. Exon 1 and part of exon 2 contain an open reading frame of 115 nt, ending one nucleotide upstream of the major reading frame. Using SOCS-1 promoter/luciferase constructs, we investigated which sequences are involved in the regulation of SOCS-1 expression. Serial promoter deletion clones indicate the localization and functionality of SP1, interferon-stimulated responsive elements (ISRE), and interferon-gamma-activated sites (GAS) promoter elements in the SOCS 1 5' flanking region. We present evidence that the upstream open reading frame (uORF) represses the translation of the downstream major open reading frame (mORF). Mutating the start codon of the uORF relieves this repression. Our data indicate that expression of the SOCS-1 protein is repressed on translational level by a mechanism, which bears similarities to that postulated for genes like retinoic acid receptor beta2 (RARbeta2), S-adenosylmethionine-decarboxylase (AdoMetDC), Bcl-2, and others. PMID- 10679191 TI - Mode of molecular recognition of L-fucose by fucose-binding legume lectins. AB - Recognition of cell surface carbohydrate moieties by lectins plays a vital role in many a biological process. Fucosyated residues are often implicated as key recognition markers in many cellular processes. In particular, the aspects of molecular recognition of fucose by fucose-bindinglectins UEA 1 and LTA pose a special case because no crystal structure of these lectins is available. The study was conducted to elucidate the process of recognition of l-fucose by UEA1 and LTA by correlating structure-based sequence alignment and other available biochemical/biophysical data. The study points out that the mode of recognition of l-fucose is coordinated by the invariant triad of residues the asparagine 137, glycine 105, and aspartate 87. The major hydrophobic stacking residue in this case is the tyrosine 220. The study also reiterates the key role of the conserved triad of residues in the combining site which is a common feature for all legume lectins whose crystal structures are known. PMID- 10679192 TI - Retinoic acid extends the in vitro life span of normal human oral keratinocytes by decreasing p16(INK4A) expression and maintaining telomerase activity. AB - Retinoic acid (RA) plays an important role in the regulation of cell growth and differentiation. To investigate whether RA extends in vitro the life span of human epithelial cells, we examined the effect of all-trans RA on both the cumulative population-doubling level (PDL) and the replicative senescence of cultured oral keratinocytes. When proliferating oral keratinocytes were cultured in medium containing 1 nM of all-trans RA, the in vitro life span of the cells was increased 1.5- to 1.8-fold compared to the vehicle control and the replicative senescence of the cells was significantly inhibited. Since the replicative senescence of human epithelial cells is associated with a steady increase of p16(INK4A) and a loss of telomerase activity, we expected that RA could delay the replicative senescence of oral keratinocytes by decreasing p16(INK4A) expression and/or inhibiting the loss of telomerase activity. To test this possibility, we examined the expression of replicative senescence-associated genes and the telomerase activities of different PDL numbers of oral keratinocytes exposed to 1 nM of all-trans RA. The protein level of cellular p16(INK4A) in the RA-treated oral keratinocytes was gradually but significantly enhanced by an increased PDL number; however, the level was significantly lower than that of the vehicle control at all of the same PDL numbers. In contrast, the telomerase activity was maintained in oral keratinocytes with increasing PDL numbers induced by RA treatment. Summarizing, these results indicate that RA induces the in vitro life-span extension of oral keratinocytes, which is linked to a decreased cellular level of p16(INK4A) and the maintenance of telomerase activity. PMID- 10679193 TI - Vomeroglandin/CRP-Ductin is strongly expressed in the glands associated with the mouse vomeronasal organ: identification and characterization of mouse vomeroglandin. AB - Vomeroglandin, a subform of mouse CRP-ductin, is a protein strongly expressed in the glands of mouse vomeronasal system. Both the proteins contain several of scavenger receptor cysteine-rich and CUB domains and one ZP domain. This domain arrangement is similar to those of rat Ebnerin, human DMBT1, and rabbit hensin. In situ hybridization analysis shows strong expression of vomeroglandin mRNA in the glands of vomeronasal system. Immunological analyses detect both membrane bound and secreted forms of vomeroglandin. The secreted protein seems to be localized in the lumen of the vomeronasal organ, playing a certain role in the pheromone perception. PMID- 10679194 TI - Isolation and characterization of dcw cluster from Streptomyces collinus producing kirromycin. AB - A 4.5-kb BamHI fragment of chromosomal DNA of Streptomyces collinus containing gene ftsZ was cloned and sequenced. Upstream of ftsZ are localized genes ftsQ, murG, and ftsW, and downstream is yfiH. Gene ftsA is not adjacent to ftsZ or other genes of the cloned fragment. Protein FtsZ was isolated and characterized with respect to its binding to GTP and GTPase activity. The binding of GTP to FtsZ was Ca(2+) or Mg(2+) dependent with an optimum at 10 mM. The rate of GTP hydrolysis by FtsZ was stimulated by KCl. The presence of Ca(2+) (3-5 mM) resulted in a significant increase of GTPase activity. Higher concentrations of Ca(2+) than 5 mM had an inhibitory effect on GTPase activity. These results indicate that divalent ions (Ca(2+) or Mg(2+)) can be involved in regulation of GTP binding and hydrolysis of FtsZ. The maximum level of FtsZ was detected in aerial mycelium when spiral loops and sporulation septa were formed. FtsZ is degraded after finishing sporulation septa. PMID- 10679195 TI - Vectors of shannon information from fourier signals characterizing base periodicity in genes and genomes. AB - Equal Symbol Fourier Transforms (FTES), characterizing nucleotide periodicity, comprise components of 5-D vectors that define base-repeat properties of a genomic sequence. This report describes a conversion of the FTES signals to a common platform of Shannon information content to facilitate comparisons of periodic data with other measures of information for genes and genomes. The autocorrelation used to compute the discrete FTES formed the basis to define repeating bases in terms of conditional probabilities. We derived a vector equation to express the Shannon information content of a sequence in a way that preserves the distinct specificity of base repeat patterns characterized by FTES vectors. We suggest application of such information vectors to study the structure of information in genes, chromosomes, and genomes by chi(2) comparisons. PMID- 10679196 TI - Purification, cloning, and three-dimensional structure prediction of Micrococcus luteus FAD-containing tyramine oxidase. AB - The FAD-containing tyramine oxidase enzyme and gene from the Gram (+) bacterium Micrococcus luteus were isolated, and computer prediction was used to propose a preliminary 3D model of the protein. A 2.8-kb Sau3AI fragment containing the structural gene of tyramine oxidase was cloned from a M. luteus genomic DNA library. The 1332 bp gene encodes a protein of 443 amino acids, with a calculated molecular mass of 49.1 kDa. The enzyme was found to be a homodimer with a molecular weight of 49,000. It oxidizes tyramine, adrenaline, 3-hydroxytyramine, dopamine, and noradrenaline, and was reversibly inhibited by FAD-containing monoamine oxidase A and B specific inhibitors. Sequence comparison show that tyramine oxidase is smaller than other FAD-amine oxidases but that it contains well-conserved amino acid residues reported in all other FAD-amine oxidases. A hypothetical three-dimensional structure of tyramine oxidase has also been proposed based on secondary structure predictions, threading, and comparative modeling. PMID- 10679197 TI - Cloning and characterization of a putative human d-2-hydroxyacid dehydrogenase in chromosome 9q. AB - There is little information on d-isomer-specific dehydrogenases in humans. Identification of d-2-hydroxyglutaric aciduria, an inherited metabolic disorder associated with severe neurological dysfunction, highlights the role of d-isomers in human metabolism. The possibility of a defect in d-2-hydroxyglutarate dehydrogenation prompted us to employ E. coli d-2-hydroxyacid dehydrogenase cDNA to search the human expressed sequence tags database. Two human EST homologues were retrieved and sequenced. Analysis showed the two clones were identical with 1258 nucleotides encoding 248 amino acids of the putative human d-2-hydroxyacid dehydrogenase. It was highly homologous to bacterial d-2-hydroxyacid dehydrogenases (46%), d-phosphoglycerate dehydrogenase (38%), and formate dehydrogenase (36%) at the amino acid level. The gene is expressed ubiquitously in tissue, most abundantly in liver, and was mapped to chromosome 9q between markers WI-3028 and WI-93330. To our knowledge this is the first cloning and characterization of the cDNA for a human d-isomer specific NAD(+)-dependent 2 hydroxyacid dehydrogenase. PMID- 10679198 TI - Deficiency of current methods in assaying endochitinase activity. AB - Since chitin is degraded by a combination of both endo- and exochitinases, it is likely that both enzymes will be present in a crude extract. Currently used substrates for detecting endochitinase activity suffer from the fact that they could easily be digested by the contaminating exochitinase, thus giving either a false-positive or an inaccurate reading of the endochitinase activity. Using Photorhabdus luminescens, a bacterium symbiotically associated with insect parasitic nematode Heterorhabditis bacteriophora as an exemplary system, we have identified these two chitinases by a simple "fluorimetric zymography" procedure. The exochitinase is a metalloenzyme and its activity is inhibited by 1,10 phenanthroline. Once the exochitinase activity is detected in a crude extract, its contribution must be eliminated before accurate determination of the endochitinase activity can be carried out. Specific properties of these enzymes including the pH activity profile, the requirement of metal ions for activity, and the molecular weight of the enzymes are among the factors to be considered in developing assaying procedures for endochitinase. PMID- 10679199 TI - Molecular characterization reveals identity of microtubule-associated proteins MAP3 and MAP4. AB - The identity of two microtubule-associated proteins, MAP3 and MAP4, was verified both immunologically and biochemically. MAP3 was enriched from the heat-stable fraction of rat brain extracts by reverse-phase HPLC and preparative two dimensional gel electrophoresis. Both MAP3 and MAP4 antibodies reacted with the corresponding spots on two-dimensional Western blots. Amino acid sequences of internal peptides derived from rat MAP3 matched with corresponding sequence stretches of mouse MAP4. In the kidney cortex, the MAP3 antibody stained not only glomerular podocytes but also interstitial cells. This distribution pattern of MAP3 is identical to that of MAP4 reported previously. These results indicate that MAP3 and MAP4 are identical. PMID- 10679200 TI - Inhibition of protein tyrosine phosphatases by low-molecular-weight S nitrosothiols and S-nitrosylated human serum albumin. AB - The homogeneous recombinant mammalian protein tyrosine phosphatase 1B (PTP1B) and Yersinia protein tyrosine phosphatase (PTPase) are inactivated by a series of low molecular-weight S-nitrosothiols. These compounds exhibited different inhibitory activities in a time- and concentration-dependent manner with second-order rate constants (k(inact)/K(I)) ranging from 37 to 113 M(-1) min(-1) against mammalian PTP1B and from 66 to 613 M(-1) min(-1) against Yersinia PTPase. Furthermore, the inactivation of Yersinia PTPase by S-nitrosylated protein:S-nitroso human serum albumin was investigated. Both single-S-nitrosylated and poly-S-nitrosylated human serum albumin show good inhibitory ability to Yersinia PTPase. The second order rate constants are 472 and 1188 M(-1) min(-1), respectively. This result indicates a possibility that S-nitrosylated albumin in vivo may function as an inhibitor for a variety of cysteine-dependent enzymes. PMID- 10679202 TI - Cloning and characterization of a novel adaptor protein, CIN85, that interacts with c-Cbl. AB - The c-Cbl protooncogene product is a prominent substrate of protein tyrosine kinases and is rapidly tyrosine-phosphorylated upon stimulation of a wide variety of cell-surface receptors. We have identified a novel c-Cbl-interacting protein termed CIN85 with a molecular mass of 85 kDa which shows similarity to adaptor proteins, CMS and CD2AP. CIN85 mRNA is expressed ubiquitously in normal human tissues and cancer cell lines analyzed. CIN85 was basally associated with c-Cbl. For interaction of CIN85 with c-Cbl, the second SH3 domain of CIN85 was shown to serve as a central player. The CIN85-c-Cbl association was enhanced shortly after stimulation of 293 cells with epidermal growth factor (EGF) and gradually diminished to a basal level, which correlated with a tyrosine phosphorylation level of c-Cbl. Our results suggest that CIN85 may play a specific role in the EGF receptor-mediated signaling cascade via its interaction with c-Cbl. PMID- 10679201 TI - PKB inhibition prevents the stimulatory effect of insulin on glucose transport and protein translocation but not the antilipolytic effect in rat adipocytes. AB - We identified 1-(5 chloronaphthalenesulfonyl)-1H-hexahydro-1, 4-diazepine, also known as ML-9, as a powerful inhibitor of PKB activity in different cells as well as of recombinant PKB. It also inhibits other downstream serine/threonine kinases, such as PKA and p90 S6 kinase, but not upstream tyrosine phosphorylation or PI3-kinase activation in response to insulin. We compared the effects of ML-9 and wortmannin on several insulin-stimulated effects in isolated rat fat cells. Both ML-9 and wortmannin inhibited glucose transport and GLUT4/IGF II receptor translocation to the plasma membrane. In contrast, only wortmannin inhibited the antilipolytic effect and PDE3B activation by insulin. Thus, ML-9 inhibits PKB but not PI3-kinase activation in response to insulin and is useful to differentiate between these effects. Both PI3-kinase and PKB are important for glucose transport and intracellular protein translocation while PKB does not appear to play an important role for the antilipolytic effect or activation of PDE3B in response to insulin. PMID- 10679203 TI - Effect of hypoxia on nitric oxide production and its synthase gene expression in rat smooth muscle cells. AB - It has not been clarified yet as to whether hypoxia and inflammation affect NO synthesis. In this study, we investigated the transcription of inducible nitric oxide synthase (iNOS) mRNA and the production of nitric oxide (NO) in rat smooth muscle cells (SMCs) cultured under hypoxic conditions in the presence and absence of proinflammatory cytokine interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS). We found that hypoxia inhibited the production of NO but did not affect the transcription of iNOS mRNA in rat SMCs treated with IFN-gamma, LPS, or both. These results indicate that O(2) is involved in the regulation of NO synthesis in inflammatory tissues. PMID- 10679204 TI - Antimicrobial peptide of korean native goat lactoferrin and identification of the part essential for this activity. AB - The antimicrobial activity of lactoferrin isolated from Korean native goat (KN goat) milk was studied and its antimicrobial domain was identified using synthetic peptides. Antimicrobial activity was assayed by a micro-method using 96 well microplates and a microplate reader. The amino acid sequence of the antimicrobial domain was suggested to be YQWQRRMRKLGAPSIT and this sequence corresponds to amino acid residues 20 to 35 of KN goat lactoferrin. Five peptides with certain amino acid residues deleted were synthesized in an effort to identify the residues essential for antimicrobial activity and it was found that the part with the sequence RRMRK (24-28) is the region most important for this activity. On the other hand, the conformation of the peptides did not influence the antimicrobial activity. PMID- 10679205 TI - Constitutive and inducible expression of hepatic CYP2E1 in leptin-deficient ob/ob mice. AB - In this study we have analyzed the inducible as well as constitutive hepatic expression of Cyp2e1 in a genetic model of obesity and non-insulin dependent (type II) diabetes, the leptin-deficient ob/ob mouse. In obese mice, Cyp2e1 levels were decreased compared to lean littermates. Treatment with leptin increased hepatic Cyp2e1 in obese mice to the levels observed in lean animals, but failed to alter Cyp2e1 expression in lean animals. As expected, leptin also reduced food intake in treated mice compared to saline-treated controls. In obese mice pair-fed the reduced amount of food, there was a significant increase in Cyp2e1 mRNA but no increase in Cyp2e1 protein or enzyme activity. Fasting and administration of acetone and 4-methylpyrazole increased Cyp2e1 mRNA as well as protein and activity in both obese and lean mice. The present data indicate that while Cyp2e1 is still inducible in obese mice by xenobiotics and fasting, full constitutive expression of Cyp2e1 requires leptin to be present. This effect of leptin appears to be at least partly independent of the hypothalamic control of food intake. PMID- 10679206 TI - Cloning and characterization of the cytochrome P450 oxidoreductase gene from the zygomycete fungus Cunninghamella. AB - The filamentous fungus Cunninghamella utilizes cytochrome P450 system(s) in the metabolism of a broad range of polyaromatic and aliphatic pollutants and a variety of drugs, but prior attempts at isolation of P450 system components of this fungus have been generally unsuccessful. We report upon the cytochrome P450 oxidoreductase (CPR) gene from two widely studied species, C. elegans and C. echinulata. The C. elegans CPR gene was obtained by screening a genomic library using as probe a PCR amplicon obtained with degenerate primers based on known CPRs. The 2420 bp coding region contained two apparent introns (149 bp and 138 bp). Northern blot analysis showed that the CPR gene is transcriptionally expressed in C. elegans and appears to be inducible by an alkane substrate, n tetradecane. Phylogenetic comparison of the deduced C. elegans CPR (710 aa) suggested that it is more closely related to animal CPRs (41-42%) than to yeast (38-41%) and plant (35-36%) forms. A 2074 bp sequence containing most of the CPR gene homolog from C. echinulata was also isolated. PMID- 10679207 TI - Alternative splicing of gar-1, a Caenorhabditis elegans G-protein-linked acetylcholine receptor gene. AB - We have recently identified a gene, designated gar-1, coding for a novel form of G-protein-linked acetylcholine (ACh) receptor in Caenorhabditis elegans. Although this receptor is most closely related to muscarinic ACh receptors (mAChRs), electrophysiological analyses have shown that ligand binding specificity of the receptor is distinct from that of mAChRs. Here we report that three receptor isoforms are generated by alternative splicing of the gar-1 transcript. These receptor isoforms differ only in the third intracellular loop that is considered to be important for G protein coupling. The three splice variants, when expressed in Xenopus oocyte, displayed similar pharmacological profiles and signaling activities. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that the three gar-1 mRNAs are present at all developmental stages examined. The results in this study provide evidence that alternative splicing is involved in promoting molecular diversity of G-protein-linked ACh receptors. PMID- 10679208 TI - Restoration of mRNA splicing by a second-site intragenic suppressor in the T4 ribonucleotide reductase (small subunit) self-splicing intron. AB - The nrdB gene of bacteriophage T4 codes for the small subunit of ribonucleotide reductase and contains a 598-base self-splicing intron which is closely related to other group I introns of T4 and eukaryotes. Thirty-one mutants causing splicing defects in the nrdB intron were isolated. Twenty-three EMS-induced revertants for these 31 primary mutants were isolated by the strategic usage of the white halo plaque phenotype. We mapped these revertants by marker rescue using subclones of the nrdB gene. Some of these second-site mutations mapped to regions currently predicted by the secondary structure model of the nrdB intron. One of these suppressor mutants (nrdB753R) was found to be intragenic by marker rescue with the whole nrdB gene. However, this mutation failed to map within the nrdB intron. Splicing assays showed that this pseudorevertant restored splicing proficiency of the nrdB primary mutation to almost wild-type conditions. This is the first example of a mutation within the exons of a gene containing a self splicing intron that is capable of restoring a self-splicing defect caused by a primary mutation within the intron. In addition, two other suppressor mutations are of interest (nrdB429R and nrdB399R). These suppressors were able to restore their primary 5' defect but in turn create a 3' splicing defect. Both of these revertants mapped in different regions of the intron with respect to their primary mutations. PMID- 10679209 TI - The metal dependence of Bacillus subtilis phytase. AB - The metal ion requirement of a Bacillus subtilis phytase has been studied. Removal of metal ions from the enzyme by EDTA resulted in complete inactivation. Circular dichroism spectroscopy was used to study the effect of metal ion removal on the protein conformation. The loss of enzymatic activity is most likely due to a conformational change, as the circular dichroism spectra of holoenzyme and metal-depleted enzyme were different. Metal-depleted enzyme was partially able to restore the active conformation when incubated in the presence of calcium. Only minor reactivation was detected with other divalent metal ions and their combinations. Based on the data we conclude that B. subtilis phytase requires calcium for active conformation. Calcium has also a strong stabilizing effect on the enzyme against thermal denaturation. However, the conformational change resulted by calcium depletion does not affect the protease susceptibility. PMID- 10679210 TI - X-Linked juvenile retinoschisis associated with a 4-base pair insertion at codon 55 of the XLRS1 gene. AB - X-linked juvenile retinoschisis (RS) is a bilateral vitreoretinal disorder with no known cure. The gene responsible for the disease was recently isolated by positional cloning methods and a spectrum of mutations has been described in families with RS pathology. In this report, we screened six sporadic cases of RS for mutations in the RS gene to understand the etiology of isolated cases. Our extensive studies revealed a novel 4 bp insertion in one family and the remaining families did not show mutations in the RS gene. This mutation altered the reading frame including codon 55 resulting in nine aberrant amino acid residues. The unaffected mother did not contain this mutation. Additionally, it was not found in 60 normal control chromosomes, suggesting that the insertion mutation is disease related in the family analyzed. PMID- 10679211 TI - Expression of the Aspergillus fumigatus phytase gene in Pichia pastoris and characterization of the recombinant enzyme. AB - Aspergillus fumigatus phytase is a heat-stable enzyme of great potential. Our objective was to determine if a high level of functional expression of the A. fumigatus phytase gene could be produced in Pichia pastoris and how the recombinant phytase reacted to different substrates, heating conditions, and proteases. A 1.4-kb DNA fragment containing the coding region of the gene was inserted into the expression vector pPICZalphaA and expressed in P. pastoris as an active, extracellular phytase (r-Afp). The yield was 729 mg of purified protein per liter of culture, with a specific activity of 43 units/mg of protein. The enzyme r-Afp shared similar pH and temperature optima, molecular size, glycosylation extent, and specificity for p-nitrophenyl phosphate and sodium phytate to those of the same enzyme expressed in A. niger. Given 20 min of exposure to 65 to 90 degrees C, the enzyme retained 20 to 39% higher residual activity in 10 and 200 mM sodium acetate than that in sodium citrate. The enzyme seemed to be resistant to pepsin digestion, but was degraded by high levels of trypsin. In conclusion, P. pastoris is a potential host to express high levels of A. fumigatus phytase and the thermostability of the recombinant enzyme is modulated by the specificity of buffer used in the heat treatment. PMID- 10679212 TI - Characterization of a new phenoloxidase inhibitor from the cuticle of Manduca sexta. AB - Melanin, the phenolic biopolymer that serves as a skin- and hair pigment protecting agent against harmful solar radiation and a free radical trap, is biosynthesized in animals mainly by the action of tyrosinase also known as phenoloxidase. Regulation of tyrosinase and hence melanogenesis is vital for all animals. In this report, we present the isolation and characterization of a new, heat-labile glycoprotein inhibitor of phenoloxidase from the larvae of Manduca sexta. The inhibitor was isolated from the live larval cuticle by buffer extraction and purified to homogeneity employing ammonium sulfate precipitation, dialysis, and concanavalin A-Sepharose chromatography. It migrated with a molecular weight of 380,000 on SDS-PAGE gels and inhibited the activity of insect and plant as well as fungal phenoloxidases. Inhibitor formed a tight complex with phenoloxidases, which resisted dissociation even by 1% Triton X-100 or SDS. Selective inhibition of phenoloxidase, while acting on certain but not all different substrates, was observed. The physiological importance of this newly discovered high-molecular-weight phenoloxidase inhibitor is discussed. PMID- 10679213 TI - Computational design of new cyclic urea inhibitors for improved binding of HIV-1 aspartic protease. AB - We report in this paper the design, by means of computational techniques, of new cyclic urea inhibitors of the HIV aspartic protease. The relationship between the complexation energies of the enzyme with known inhibitors and the experimentally determined log K(i) have been studied and used to predict inhibition constants for new inhibitors. PMID- 10679214 TI - Light chain of natural antibody plays a dominant role in protein antigen binding. AB - Examinations of the contribution and the specificity of heavy (H) and light (L) chains of natural antibodies to antigen binding may help us to better understand antigen recognition and the development of naive B cells. We previously generated natural Fab antibody fragments reactive to preS1 of HBV using a naive, non immunized Fab antibody library derived from peripheral B cells of a normal healthy volunteer. We now constructed expression vectors for the Fd (VH + CH1), L chain, and scFv fragments using the sequences encoding parental Fabs as a source of natural antibody genes. The recombinant antibody fragments were expressed as inclusion bodies in E. coli BL21 (DE) cells. When denatured and then refolded, the antibody fragments retained their binding properties. Recombinant L chains and scFvs exhibited three- to 40-fold higher affinities (in the order of 10(7) M( 1)) over the parental Fabs, whereas the affinities of Fds (in the order of 10(5) M(-1)) were much lower compared to the parental Fabs. The results obtained from sandwich ELISA revealed that the L chains bound the virus more efficiently than Fds. Additional experiments were performed to evaluate the specificity of the recombinant fragments for surface proteins of HBV. Fds and L chains were reactive towards HBsAg and the preS2 peptide as well as preS1 and showed patterns of epitope recognition quite different from those of parental Fabs. The data presented here demonstrate that the prominence of the L chain in determining protein binding activity is a property of natural antibodies and is quite unlike the antibodies induced by immunization, and that the specificity of Fab is not determined by the individual antibody chain but by the correct pairing of H and L chain. PMID- 10679215 TI - Interactions of cytochrome c peroxidase with lysine peptides. AB - Structural change of Cytochrome c peroxidase (CcP) due to interaction with lysine peptides (Lysptds) has been studied by absorption spectra and measurements on electron transfer between cytochrome c (cyt c) and CcP in the presence of Lysptd. Peaks were observed in the difference absorption spectrum of CcP between in the presence and absence of Lysptds, demonstrating a structural perturbation of CcP, at least at its heme site, on interaction with Lysptd. The interaction between CcP and Lysptd was electrostatic, since no significant peak was detected in the difference absorption spectrum when 100 mM of NaCl was added to the solution. Lysptds competitively inhibited electron transfer from cyt c to CcP, which indicated that they interacted with CcP at the same site as cyt c and would be models of the CcP interacting site of cyt c. PMID- 10679216 TI - High affinity ScFvs from a single rabbit immunized with multiple haptens. AB - We report the generation of single-chain Fv (scFv) fragments with high affinities against four different hapten molecules from a single immunised rabbit. The rabbit was immunised with a mixture of protein conjugates of four different haptens, namely the herbicide mecoprop and derivatives of the herbicides atrazine, simazine, and isoproturon. An scFv phage display library was constructed, and several scFvs with high affinity against each hapten were isolated. For each hapten, a single binder was selected by k(off) ranking and used for affinity determination. The affinities were in sub-nanomolar range and the lowest K(d) value obtained was 6.75 x 10(-10) M. An unusual feature of one of the anti-isoproturon scFvs was its ability to retain binding activity at pH1.7. The utility and potential of using a single animal and immunisation with multiple antigens for the production of multiple, specific, high affinity scFvs by phage display is discussed. PMID- 10679217 TI - A novel function of serum amyloid A: a potent stimulus for the release of tumor necrosis factor-alpha, interleukin-1beta, and interleukin-8 by human blood neutrophil. AB - High density lipoprotein (HDL) and its main apolipoproteins, AI and serum amyloid A (SAA), present in physiological and acute phase response conditions, respectively, affect the inflammatory process. This study focuses on the effect of AI, SAA, and HDL from healthy (N-HDL) and acute phase individuals (AP-HDL) on the release of TNF-alpha, IL-1beta, and IL-8 by human blood neutrophils. It was observed that SAA (100 microg/mL) causes a dramatic increase (75-400 times) in the basal liberation of the three cytokines assayed. This effect is not triggered by AP-HDL. Although AI (100 microg/ml) increases the release of IL-1beta and IL-8 modestly, N-HDL does not. Both HDLs (0.16-0.32 mg of protein/mL) had an anti inflammatory action, decreasing the basal and LPS-stimulated cytokine release. Given that the biological role of SAA is still uncertain, the present study adds an important finding potentially pertinent to the biological role of this acute phase protein. PMID- 10679218 TI - Physiological evidence for an interaction between helix XI and helices I, II, and V in the melibiose carrier of Escherichia coli. AB - In a previous study 23 residues in helix XI of the cysteine-less melibiose carrier were changed individually to cysteine. Several of these cysteine mutants (K377C, A383C, F385C, L391C, G395C) had low transport activity and they were white on melibiose MacConkey fermentation plates. After several days of incubation of these white clones on melibiose MacConkey plates a rare red mutant appeared. The plasmid DNA was then isolated and sequenced. The two second site revertants from K377C were I22S and D59A. This change of aspartic acid to a neutral residue suggests that physiologically there is an interaction between K377 and D59 (possibly a salt bridge). The revertants from A383C were in positions 20 (F20L) and 22 (I22S and I22N). Revertants of F385C were intrahelical changes (I387M and A388G) and a change in C-terminal loop (R441C). Revertants of L391C were in helix I (I22N, I22T and D19E) and helix V (A152S). Revertants of G395C were in helix I (D19E and I22N). We suggest that there is an interaction between helix XI and helices I, II, and V and proximity between these helices. PMID- 10679219 TI - Differential stimulation of prostaglandin G/H synthase-2 in osteocytes and other osteogenic cells by pulsating fluid flow. AB - Mechanical stress produces flow of fluid in the osteocytic lacunar-canalicular network, which is likely the physiological signal for the adaptive response of bone. We compared the induction of prostaglandin G/H synthase-2 (PGHS-2) by pulsating fluid flow (PFF) and serum in osteocytes, osteoblasts, and periosteal fibroblasts, isolated from 18-day-old fetal chicken calvariae. A serum-deprived mixed population of primarily osteocytes and osteoblasts responded to serum with a two- to threefold induction of PGHS-2 mRNA. Serum stimulated PGHS-2-derived PGE(2) release from osteoblasts and osteocytes but not from periosteal fibroblasts as NS-398, a PGHS-2 blocker, inhibited PGE(2) release from osteocytes and osteoblasts with 65%, but not that from periosteal fibroblasts. On the other hand PFF (0.7 Pa, 5 Hz) stimulated (3 fold) PGHS-2 mRNA only in OCY. The related PGE(2) response could be completely inhibited by NS-398. We conclude that osteocytes have a higher intrinsic sensitivity for loading-derived fluid flow than osteoblasts or periosteal fibroblasts. PMID- 10679220 TI - Characterization of Erwinia chrysanthemi PY35 cel and pel gene existing in tandem and rapid identification of their gene products. AB - Genomic DNA of the phytopathogenic Erwinia chrysanthemi PY35 was partially digested with Sau3AI, ligated into the BamHI site of pBluescript II SK+, and introduced into E. coli. One clone that was able to hydrolyse carboxymethylcellulose and polygalacturonic acid was selected. A 2.9 kb fragment containing the pelL1 gene (pPY300) and cel5Z gene (pPY401) in tandem was subcloned and sequenced. The pelL1 and cel5Z genes had open reading frames of 1,278 bp and 1,281 bp encoding 425 and 426 amino acid residues with calculated molecular weights of 45,649 Da and 46,473 Da, respectively. pelL1 and cel5Z carried a typical prokaryotic signal peptide of 24 and 41 amino acid residues, respectively. The apparent molecular masses of the proteins when expressed in E. coli cells were approximately 43 kDa (PelL1) and 42 kDa (Cel5Z) as assessed by PGA-SDS-PAGE and CMC-SDS-PAGE. PMID- 10679221 TI - Correlation between the chaperone-like activity and aggregate size of alpha crystallin with increasing temperature. AB - alpha-Crystallin, the major protein of the mammalian eye lens, is also found in the major tissues of the body, where one or the other of its two isoforms is characteristically expressed. Both isoform sequences are highly related to others of the small heat shock protein superfamily, leading to speculation about their functions in vivo outside of the lens. Tests of chaperone-like activity at 37 and 66 degrees C indicate that the protein can act to prevent the superaggregation of partially denatured proteins, but both alpha-crystallin aggregate size and shape are significantly altered with increasing temperature. Characterization of these changes indicates that secondary, tertiary, and quaternary structure are modified, with the latter effect especially striking above 50 degrees C. Furthermore, these changes appear to be irreversible when the temperature is returned to 25 or 37 degrees C. Functionally, the protein is effective in chaperone-like activity at all temperatures, but exhibits a somewhat increased capability after a cycle of heating and cooling. The results presented here indicate the heat-induced formation of high-molecular-weight aggregates of alpha crystallin is a slow progressive process. The increased activity of these aggregates suggests that chaperone-like activity depends in part on the packing parameters of the aggregate and on conformation of the subunit within that aggregate. PMID- 10679222 TI - Kassinatuerin-1: a peptide with broad-spectrum antimicrobial activity isolated from the skin of the hyperoliid frog, Kassina senegalensis. AB - Kassinatuerin-1 (GFMKYIGPLI(10)PHAVKAISDL(20)I.NH(2)) was isolated in high yield (75 nmol/g) from an extract of the skin of a Hyperoliid frog, the African running frog Kassina senegalensis and its sequence was confirmed by total synthesis. The peptide inhibited growth of the gram-negative bacterium Escherichia coli (minimum inhibitory concentration, MIC = 4 microM), the gram-positive bacterium Staphylococcus aureus (MIC = 8 microM), and the yeast Candida albicans (MIC = 70 microM). A structurally related peptide, kassinatuerin-2 (FIQYLAPLI(10)PHAVKAISDL(20)I.NH(2)) was also isolated in high yield (96 nmol/g) from the extract but was devoid of antimicrobial activity against these microrganisms. Kassinatuerin-1 may be classified with other linear, cationic antimicrobial peptides that can potentially adopt an amphipathic alpha-helical conformation but it contains almost no amino acid sequence identity with previously characterized bioactive peptides from frog skin. PMID- 10679223 TI - Human 3'-phosphoadenosine 5'-phosphosulfate synthetase 1 (PAPSS1) and PAPSS2: gene cloning, characterization and chromosomal localization. AB - Sulfae conjugation is an important pathway in the metabolism of a large number of exogenous and endogenous compounds. These reactions are catalyzed by sulfotransferase (SULT) enzymes that utilize 3'-phosphoadenosine 5' phosphosulfate (PAPS) as a sulfate donor. PAPS is synthesized from ATP and inorganic sulfate by PAPS synthetase (PAPSS). Two separate PAPSS cDNAs, PAPSS1 and PAPSS2, have been identified in human tissues. We have cloned and characterized the genes for human PAPSS1 and PAPSS2 to make it possible to study the pharmacogenomics of these enzymes. Both genes consisted of 12 exons with virtually identical exon-intron splice junction locations. All splice junctions conformed to the "GT-AG" rule. The total length of PAPSS1 was approximately 108 kb, while that of PAPSS2 was greater than 37 kb. The 5'-flanking region of PAPSS1 did not include a TATA box sequence near the site of transcription initiation, but PAPSS2 had a TATA motif located 21 bp upstream from the site of transcription initiation. Northern blot analysis showed that the major PAPSS1 and PAPSS2 transcripts were approximately 2.7 and 4.2 kb in length, respectively. PAPSS1 mapped to human chromosome band 4q24 while PAPSS2 mapped to 10q22-23 by fluorescence in situ hybridization analysis. Cloning and structural characterization of PAPSS1 and PAPSS2 will make it possible to perform molecular genetic and pharmacogenomic studies of these important enzymes in humans. PMID- 10679224 TI - Number of glomeruli is increased in the kidney of transgenic mice expressing the truncated type II activin receptor. AB - Histological analyses of the kidney were performed in transgenic mice expressing the truncated type II activin receptor. In these mice, signaling through the activin receptor was attenuated. Size and wet weight of the kidneys were identical to those of normal mice. Histologically, the number of glomeruli was approximately 180% of that in normal mice. The sizes and shapes of the glomeruli were variable, but many of them were smaller than those in normal mice. Morphometrically, the total glomerular area was 130% of that of the normal mice. Abnormality of the epithelia in Bowman's capsule was observed and the number of tubular epithelial cells was increased in the transgenic mice. The serum levels of blood urea nitrogen, creatinine, and creatinine clearance were identical to those in normal mice. These results suggest that the action of activin or related ligands is critical for determination of the nephron number. PMID- 10679225 TI - Multipotency of a bone marrow stromal cell line, TBR31-2, established from ts SV40 T antigen gene transgenic mice. AB - Bone marrow is believed to contain multipotential stromal stem cells which can differentiate into osteoblasts, chondrocytes, adipocytes, and myoblasts (Prockop, D. J. Science 276, 71-74, 1997). Therefore, characterization and identification of the stem-like cell within the stromal cells are important to understand bone marrow function in relation to the hematopoietic microenvironment, and repair/regeneration of tissue defects. TBR31-2 cell, a bone marrow stromal cell line established from bone marrow of transgenic mice harboring temperature sensitive (ts) simian virus (SV) 40T-antigen gene for immortality, is induced toward both adipocytic and osteogenic cells under conditions of the inactivation of T-antigen (Okuyama, R., Yanai, N., Obinata, M. Exp. Cell Res. 218, 424-429, 1995). In this work, using a semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, mRNA expressions of tissue-specific differentiation markers for adipocyte (lipoprotein lipase), osteoblast (type I collagen and osteocalcin), chondrocyte (type II and X collagen), and muscle cell (desmin) were examined during a long-term culture of the cell. In addition, histochemical studies showed the appearance of adipocytic, osteoblastic, chondrocytic, and muscle cells during this long-term culture. Thus, TBR31-2, which has characteristics of an undifferentiated cell, has the potential to express the multipotential cell lineages. These results indicated that a multipotential progenitor cell including potential to differentiate into a muscle cell and which is situated in the mesenchymal cell lineage was first obtained. PMID- 10679226 TI - Human hepatitis B virus X protein is a possible mediator of hypoxia-induced angiogenesis in hepatocarcinogenesis. AB - The hepatitis B virus (HBV)-encoded transcriptional activator HBV-X protein (HBx) was known to be involved in hepatocarcinogenesis. Hepatocarcinogenesis generally included an active angiogenesis that was mainly considered to be due to a local hypoxia in liver tissues. However, the exact mechanisms of HBx-induced hepatocarcinogenesis were poorly understood. In this study, we examined the role of HBx in the increased angiogenesis and the possible regulating mechanisms of HBx by hypoxia. We demonstrated that HBx stimulated the transcription of vascular endothelial growth factor (VEGF), a potent angiogenic factor, in HBx-stable transfectants. HBx-induced angiogenesis was confirmed by in vivo tumor angiogenesis assay, resulting in that the HBx transfectants increased the formation of new blood vessels compared to the control transfectants. Then, we demonstrated that the expression of HBx was enhanced after incubating HBV infected hepatoma cells under hypoxia. Moreover, the activity of HBV enhancer 1 (Enh1) was increased when hepatoma cells transfected with the reporter plasmid containing HBV Enh1 were exposed to hypoxic conditions. These results strongly suggest that HBx may play a critical role in the hypoxia-induced angiogenesis through transcriptional activation of VEGF during hepatocarcinogenesis. PMID- 10679227 TI - Engineered metal binding sites on green fluorescence protein. AB - The ability to assay a variety of metals by noninvasive methods has applications in both biomedical and environmental research. Green fluorescent protein (GFP) is a protein isolated from coelenterates that exhibits spontaneous fluorescence. GFP does not require any exogenous cofactors for fluorescence, and can be easily appended to other proteins at the DNA level, producing a fluorescence-labeled target protein in vivo. Metals in close proximity to chromophores are known to quench fluorescence in a distance-dependent fashion. Potential metal binding sites on the surface of GFP have been identified and mutant proteins have been designed, created, and characterized. These metal-binding mutants of GFP exhibit fluorescence quenching at lower transition metal ion concentrations than those of the wild-type protein. These GFP mutants represent a new class of protein-based metal sensors. PMID- 10679228 TI - Characterization of a novel gene encoding a phytocyanin-related protein in morning glory (Pharbitis nil). AB - A cDNA for a novel phytocyanin homolog was cloned from apical buds of morning glory (Pharbitis nil). The predicted protein was most similar to a family of early nodulins, which are expressed during the formation of symbiotic root nodules of legume plants, and less similar to typical phytocyanins such as lacquer tree stellacyanin and cucumber basic protein. The amino acid sequence predicted that it is a secreted protein associated with other components of the extracellular matrix. Hybridization analyses showed that the transcript was expressed specifically in meristems and procambia of apical buds and root tips. The transcript level in the apical buds decreased significantly on flower inducing treatment. Involvement of this phytocyanin-related protein in plant organ differentiation is discussed. PMID- 10679229 TI - Enhancement of the migration of metastatic human breast cancer cells by phosphatidic acid. AB - Phosphatidic acid (PA), lysophosphatidic acid (LPA), and sphingosine 1-phosphate (SPP) are naturally occurring phospholipids which induce a variety of effects as extracellular messengers. In this study, we compared the effects of these phospholipid signaling molecules on the migration of invasive and noninvasive breast cancer cell lines, an index of the metastatic potential of these cells. As previously demonstrated, invasive MDA-MB-231 breast cancer cells exhibited increased constitutive (nonstimulated) migration in comparison to poorly invasive MCF-7 cells. Phosphatidic acid employed at nanomolar concentrations markedly potentiated migration of the invasive cells but had no effect on migration of either the noninvasive MCF-7 cells or nonneoplastic human epithelial cells. Lysophosphatidic acid and sphingosine 1-phosphate inhibited both the directed (chemotactic) and random (chemokinetic) migration of MDA-MB-231 cells. Experiments were undertaken to characterize the signaling pathway involved in constitutive and PA-stimulated migration of MDA-MB-231 cells. The tyrosine kinase inhibitors staurosporine and genistein inhibited constitutive and PA-induced migration in a dose-dependent manner, consistent with a role for tyrosine phosphorylation in the migratory response. In addition, the phosphatidylinositol (PI) 3' kinase inhibitors wortmannin and LY294002 strongly inhibited both the constitutive and PA-stimulated migration of the invasive breast cancer cells, indicating that PI-3' kinase plays an important role in the metastatic migration of breast cancer cells. Finally, PA-induced migration of MDA-MB-231 was markedly attenuated by pretreatment of cells with Clostridium difficile Toxin B, pertussis toxin and suramin, implying a role for a Gi receptor-dependent process involving activation of the small GTP-binding protein Rho. Since an enhanced ability to migrate heightens the metastatic potential of cells within solid tumors, our results suggest that the metastatic capabilities of breast cancer cells may be enhanced by a receptor-driven cellular process initiated by phosphatidic acid or related lipid phosphate messengers. PMID- 10679230 TI - Nicotine binding to native and substituted peptides comprising residues 188-207 of nicotinic acetylcholine receptor alpha1, alpha2, alpha3, alpha4, alpha5, and alpha7 subunits. AB - Structural determinants of L-[(3)H]nicotine binding to synthetic peptides comprising residues 188-207 of nicotinic acetylcholine receptor alpha subunits were invesitigated by equilibrium binding analysis. Two binding components were detected, one of low affinity (K(d) approximately 1.5 microM) that did not differ significantly among peptides and another of high affinity. The high affinity binding component was higher for the neuronal peptides (K(d) = 14-23 nM) than the muscle alpha1 peptides (K(d) = 52 nM). The following nonconservative substitutions in the alpha4 peptide resulted in a significant decrease in nicotine affinity for the peptide: Y190A, Y190D, C192G, E195A, E195-, P199A, P199 , and Y203A. Substitution of alpha4P199 with a leucine which is present in the alpha1 sequence decreased the affinity of the alpha4 peptide for nicotine and substitution of alpha1L199 with a proline (alpha4) or a glutamine (alpha3) increased the affinity of the alpha1 peptide. It is concluded that aromatic residues contribute to the binding site for nicotine on the alpha4 subunit and that the residue present at position 199 partly determines differences in nicotine affinity for different alpha subunits. PMID- 10679231 TI - Pancreatic acinar cells submitted to stress activate TNF-alpha gene expression. AB - To elucidate whether pancreatic acinar cell submitted to stress is able to express TNF-alpha, we studied TNF-alpha mRNA expression by Northern blot and in situ hybridization in healthy pancreas, in tissue from caerulein-induced pancreatitis and after lipopolysaccharide (LPS) treatment. In specimens from normal pancreas, TNF-alpha mRNA expression, as judged by both Northern blot and in situ hybridization, was negative, whereas a strong but transient expression was observed in acinar cells from caerulein pancreatitis and LPS treatment. TNF alpha mRNA appeared as rapidly as 30 min after treatment, and was maximal 6 h after. At this time, there was mild infiltration consisting mostly of polymorphonuclear leukocytes (PMNL) and no signal of TNF-alpha transcript was found in their cytoplasm. Our results strongly indicate that pancreatic acinar cell is the source of TNF-alpha early in the course of acute pancreatitis and LPS treatment, and suggest that the expression of this cytokine is a part of a general response of the acinar cell to aggression. PMID- 10679232 TI - The role of glucose and its metabolism in the regulation of glucokinase expression in isolated human pancreatic islets. AB - Previous reports concerning the regulation of glucokinase expression in beta cells have been done using cell models from rodent origin. Evidence is lacking so far to implicate the same regulatory mechanisms in human cells. In this study, we investigate the effects of glucose on the expression of glucokinase using isolated human pancreatic islets. High glucose (16.7 mM), in a time-dependent manner, increases the amount of immunoreactive glucokinase (+150% after 7 days culture, P < 0.01) without apparent changes in glucokinase gene expression, suggesting that glucose exerts its effect at a posttranscriptional level. Mannose, but not the nonmetabolized hexoses, 3-O-methylglucose or 2-deoxyglucose, increases glucokinase protein content. Even though these findings are compatible with an involvement of signals derived from glucose metabolism, additional data argue against this hypothesis: (i) a glucokinase inhibitor (mannoheptulose) does not block glucose-induced increase in glucokinase content and (ii) other metabolic fuels (amino acids) are ineffective. We suggest that the glucose molecule, by mechanisms yet to be defined, but probably not involving its metabolism, regulates human glucokinase expression. PMID- 10679233 TI - Peridinin as the major biological carotenoid quencher of singlet oxygen in marine algae Gonyaulax polyedra. AB - Carotenoids in light-harvesting proteins and reaction centers increase the overall efficiency of photosynthesis by transferring absorbed light energy to chlorophylls. Peridinin and beta-carotene were isolated from Gonyaulax polyedra in a one-step purification protocol using the preparative circular chromatography (Chromatotron), performed on silica gel under N(2) atmosphere and n hexane/acetone 8:2 as mobile phase and characterized by extensive (1)H NMR, infrared, and electrospray ionization mass spectrometry analyses. The quenching of singlet molecular oxygen [O(2) ((1)Delta(g))] was evaluated by NIR-emission assays using singlet oxygen generated by sensitization of either perinaphthenone or methylene blue. The NIR-emission assay showed that peridinin quench as singlet oxygen (k(q) = 9.5 x 10(8) M(-1) s(-1)) 5-fold less efficiently than beta carotene (52 x 10(8) M(-1) s(-1)). A method, based on the use of high-performance liquid chromatography with UV-VIS detection, was then developed for the sensitive quantification of peridinin (55% of total carotenoids) and beta-carotene (4.1% of total carotenoids). Thus, since peridinin is 10-fold more abundant than beta carotene, it is expected to be the major protector against the deleterious effects of O(2) ((1)Delta(g)) in Gonyaulax polyedra. PMID- 10679234 TI - Histone H1 variants play individual roles in transcription regulation in the DT40 chicken B cell line. AB - Thirty-nine of the 44 chicken histone genes are located in a major gene cluster of 110 kb, the others being distributed in four separate regions. All 6 H1 genes, which are present in the cluster and encode different variants, are expressed in the DT40 chicken B cell line, at levels ranging from about 5 to 40%. To clarify differences in the natures of these H1 variants, using gene-targeting techniques, we generated a series of DT40 mutants, which are devoid of each of the 5 H1 genes, respectively. Analyses of six H1-deficient mutants, comprising the latter five and a previously generated H1-deficient mutant, revealed that the protein patterns on 2D-PAGE were definitely different from each other, indicating that each H1 variant plays an individual role in the transcription regulation of specific genes in DT40 cells. PMID- 10679235 TI - Identification of a nuclear ribonucleoprotein particle which contains incompletely spliced HIV-1 RNAs. AB - Unspliced and partially spliced HIV RNAs are transported to the cytoplasm by the HIV encoded Rev protein. In the present study, a ribonucleoprotein complex which contains such incompletely spliced HIV RNA is identified. Soluble nuclear extracts were prepared from the lymphocyte cell line H9/IIIB that constitutively produces HIV-1 from a stably integrated provirus. Sucrose gradient centrifugation of the extracts and subsequent analysis of the gradient fractions by a ribonuclease protection assay revealed a population of incompletely spliced HIV-1 RNAs which accumulates in the 100S region of the gradient. Similar analysis of cellular mRNAs including beta-actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) revealed that these RNA molecules also exhibit characteristic sedimentation profiles in sucrose gradients. This study suggests that nuclear ribonucleoprotein particles containing incompletely spliced HIV-1 RNAs are amenable for biochemical characterisation. PMID- 10679236 TI - Activation of caspase-3 in molt4 cells by apoptosis-inducing nucleosides from CD57(+)HLA-DR(bright) natural suppressor cell line. AB - Apoptosis-inducing nucleosides (AINs), which were released and isolated from CD57(+)HLA-DR(bright) natural suppressor (57.DR-NS) cell line derived from human decidual tissue, induced apoptosis in Molt4 cells. The addition of caspase-3 inhibitor into the reaction blocked the cleavage of caspase-3 and apoptosis in Molt4 cells treated with AINs, detected by flow cytometrical or spectrofluorometrical analysis and DNA fragmentation assay. Furthermore, by means of immunoblotting, the processing of caspase-3 was shown with the appearance of their catalytically active subunits of 20 and 11 kDa during the generation of apoptosis in Molt4 cells treated with AINs. This processing of caspase-3 into active subunits was also blocked by the addition of caspase-3 inhibitor. Thus, it was definitely revealed that the activation of caspase-3 was a key feature in the caspase cascade of AINs-induced apoptosis in Molt4 cells. PMID- 10679237 TI - Duality monomer-dimer of the pheromone-binding protein from Bombyx mori. AB - The analysis of a recombinant pheromone-binding protein from the silkworm moth, Bombyx mori, by native gel electrophoresis with Coomassie staining showed one single band with a molecular mass consistent with a monomer. A slow migrating band, detected in the recombinant and native samples by a polyclonal antibody, was indistinguishable from the monomer in the mass spectrum fragmentation pattern and chromatographic behavior. Flow injection analyses of the protein by mass spectrometry in the negative mode showed fragments of a dimer. The dimeric form was also supported by estimation of the molecular mass by gel filtration at basic pH. A cross-linked dimer coeluted with the noncovalent dimer on a gel filtration column. The molecular mass of the protein changed in a pH-dependent way with a dramatic transition from dimer to monomer between pH 6 and 4.5. A low pH induced not only dissociation of the dimer, but also a conformational change in the protein. In marked contrast to denaturation with guanidinium chloride, the emission maxima of tryptophan was not significantly changed at low pH. BmPBP is thus a dimer at slightly acid, neutral, and basic pH, which dissociates and then undergoes conformational change at low pH. PMID- 10679238 TI - Human p55(CDC)/Cdc20 associates with cyclin A and is phosphorylated by the cyclin A-Cdk2 complex. AB - The initiation of anaphase and exit from mitosis depend on the activation of the anaphase-promoting complex/cyclosome (APC/C), a multicomponent, ubiquitin-protein ligase. The WD-repeat protein called p55(CDC)(Cdc20) directly binds to and activates APC/C. By using yeast two-hybrid screening, we found that cyclin A, a critical cell cycle regulator in the S and G2/M phases, specifically interacts with p55(CDC). Ectopically expressed p55(CDC) and cyclin A form a stable protein complex in mammalian cells. The p55(CDC)-cyclin A interaction occurs through the region containing the WD repeats of p55(CDC) and the region between the destruction box and the cyclin box of cyclin A. In addition to the physical interaction, p55(CDC) is phosphorylated by cyclin A-associated kinase. These findings suggest that the function of p55(CDC) is mediated or regulated by its complex formation with cyclin A. PMID- 10679239 TI - Genome-wide detection of unknown subtle mutations in bacteria by combination of MutS and RDA. AB - We propose a procedure for detecting unknown, subtle DNA changes throughout the entire bacterial genome by a combination of MutS and RDA. Current techniques detect subtle mutations after PCR amplification of the target regions, so the mutation detection is done between amplified PCR fragments. In this paper, genome wide subtle mutation scanning in bacteria was performed by combining the MutS and RDA techniques. Our strategy for cloning a small mutation region is composed of two steps: an enrichment of fragments containing subtle mutations using MutS, followed by an RDA subtraction procedure for further enrichment. We successfully identified small mutations such as a four-base insertion, a two-base insertion, and transition mutations in bacteria. PMID- 10679240 TI - Signaling through the p38 and p42/44 mitogen-activated families of protein kinases in pancreatic beta-cell proliferation. AB - The present study has focused on the role of the 42- and 44-kDa mitogen-activated protein kinases (p42/44 MAPKs) and the 38-kDa mitogen-activated protein kinase (p38 MAPK) in the proliferation of the pancreatic beta-cell line MIN6. MIN6 beta cell proliferation was assessed by measuring 5-bromo-2'-deoxyuridine (BrdU) incorporation into cellular DNA. Inhibition of both the p42/44 MAPK pathway using the MEK inhibitor PD098059 (PD) and the p38 MAPK pathway using the p38 inhibitor SB203580 (SB) caused a marked, concentration-dependent reduction in the BrdU immunostaining observed in the presence of 15% FCS when assessed using fluorescence immunocytochemistry. These data provide direct evidence of a role for p42/44 MAPKs in the mitogenic response of MIN6 beta-cells to FCS. Furthermore, these data also suggest a novel role for the p38 MAPK pathway in MIN6 beta-cell proliferation. PMID- 10679241 TI - E-Selectin expression in a murine model of chronic colitis. AB - The objective of this study was to quantify E-selectin surface expression in the colon as well as other tissues in a CD4(+) T-cell model of chronic colitis in mice using the newly developed dual radiolabel monoclonal antibody technique. Male SCID mice were reconstituted with either 5 x 10(5) CD4(+) CD45RB(low) or CD45RB(high) T-cells isolated from normal CB-17 donor mouse spleens and subsequently monitored for clinical signs of colitis. We found that animals injected with CD45RB(high) but not CD45RB(low) T-cells nor PBS developed colitis at 6-8 weeks following reconstitution as assessed by loss of body weight, development of loose stools and/or diarrhea, and histopathology. Concurrent with the onset of distal bowel inflammation was enhanced expression of E-selectin compared to SCID mice injected with PBS or reconstituted with CD45RB(low) T cells, both of which did not develop colitis. We also observed significant increases in E-selectin expression in cecum, small intestine, mesentery, and liver of colitic mice. Our data confirm that reconstitution of SCID mice with CD45RB(high) but not CD45RB(low) T-cells induces chronic colitis and demonstrate that this chronic colitis is associated with enhanced expression of an endothelial cell-specific adhesion molecule. Furthermore, our studies demonstrate that reconstitution of SCID mice with CD45RB(high) T-cells enhances E-selectin expression in a variety of tissues distant from the site of active inflammation. PMID- 10679242 TI - Growth suppression of Escherichia coli by induction of expression of mammalian genes with transmembrane or ATPase domains. AB - Growth inhibition of Escherichia coli host cells is frequently observed when some mammalian genes are induced to express exogenously. To find common features of these mammalian genes, an assay was designed for the isolation of these genes which show growth-inhibitory effect on E. coli by induction of expression. Of 38,000 clones derived from a mouse brain cDNA library, 64 cDNA clones were systematically selected out by this method, of which 45 clones had putative open reading frames encoding proteins with putative membrane-associated regions or ATP binding/ATPase activities. These results show that a fraction of membrane associated proteins or ATP-binding/ATPase genes can be isolated from cDNA libraries by our simple method. PMID- 10679243 TI - Identification of casein kinase I substrates by in vitro expression cloning screening. AB - Casein kinase I (CKI) is a widely expressed protein kinase family implicated in diverse processes including membrane trafficking, DNA repair, and circadian rhythm. Despite the large number of CKI genes, few biologically relevant substrates have been identified. As an approach to better defining the spectrum of CKI substrates, we extended a recently described in vitro expression cloning (IVEC) strategy. Polypeptides pools were screened for kinase-dependent electrophoretic mobility shifts. Ten putative CKI substrates were isolated from an initial sample of 3000 random cDNA clones. Candidate substrates include proteins involved in RNA metabolism (a putative RNA helicase, the nucleolar protein hNOP56, and hnRNP A1, and ribosomal proteins L4, L8, and L13), as well as keratin 17, a necdin-related protein, and the calcium-binding proteins desmoglein 2 and annexin II. The same pools were also screened with active ERK2, and four substrates identified: aldolase, NSD-like protein, uracil-DNA glycosylase, and HHR23A. IVEC is an effective method to identify novel protein kinase substrates. PMID- 10679244 TI - Effect of somatostatin and octreotide on proliferation and vascular endothelial growth factor secretion from murine endothelial cell line (HECa10) culture. AB - Angiogenesis, development of new blood vessels, is required for normal tissue repair and also for tumor cell proliferation, extracellular matrix invasion, and hematogenous metastases. Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that has been shown to play a key role in neovascularization. Inhibition of angiogenesis in vitro and in vivo was documented by administration of native neuropeptide somatostatin and its analog octreotide. We have studied the effect of somatostatin-14 (SRIF) and ocreotide (sandostatin) on proliferation activity and VEGF release from cultured murine endothelial cells HECa10 in vitro. SRIF in concentrations from 10(-9) to 10(-5) M and ocreotide in concentrations from 10(-9) to 10(-5) M diminished the proliferative activity of cultured cells vs controls. SRIF and ocreotide in concentrations from 10(-14) to 10(-6) M did not change the release of VEGF into supernatants of 24 or 72 h endothelial cell cultures. Although we showed the antiproliferative effect of SRIF and ocreotide on mouse endothelial cells, we were unable to demonstrate the inhibitory effect of tested peptides on VEGF secretion in vitro. PMID- 10679245 TI - Rapid agonist-induced phosphorylation of the human CRF receptor, type 1: a potential mechanism for homologous desensitization. AB - Agonist-induced phosphorylation of the human corticotropin-releasing factor type 1 receptor (hCRF(1)-R) was investigated using an influenza hemagglutinin (HA) epitope-tagged receptor transiently expressed in COS-7 cells. The HA-hCRF(1)-R migrated as a broad band (M(r) 60,000-70,000) in SDS-PAGE and showed increased mobility (M(r) approximately 48,000) after enzymatic deglycosylation with peptide N-glycosidase F, consistent with the predicted size (47 kDa) of the nonglycosylated HA-hCRF(1)-R protein. A marked increase in HA-hCRF(1)-R phosphorylation was observed in HA-hCRF(1)-R-expressing COS-7 cells exposed to 1 microM ovine CRF for 5 min, whereas activation of protein kinase A (PKA) by 50 microM forskolin, or of Ca(2+)/calmodulin (CaM)-dependent kinases by 10 microM ionomycin, had little effect. These findings are consistent with preliminary data suggesting that CRF(1)-R phosphorylation mediated by G protein receptor kinase 3 (GRK3), but not by PKA or CaM-dependent kinases, has an important role in the homologous desensitization of brain CRF(1)-Rs. PMID- 10679246 TI - Characterization of a postreceptor signaling defect that impairs cfos expression in cultured fibroblasts of a patient with insulin resistance. AB - Induction of cfos expression is a definite end point of signal transduction by receptor tyrosine kinases via MAPK cascades. We have examined signal transduction to transcription factor cFos in isolated fibroblasts of a patient with an inherited syndrome of insulin resistance. MAPK phosphorylation and activity were unaltered, but inducibility of cfos transcription was strongly impaired by insulin and reduced by PDGF. Induction of the cfos promoter via MAPK is mediated by activation of the ternary complex. Abundance of SRF or Elk-1 was unaltered, but Elk-1 phosphorylation following stimulation was reduced. Transient transfections with reporter genes under control of the Elk-1 binding ets/sre cis element or expression plasmids coding for the regulatory domain of Elk-1 fused to heterologous DNA binding domains revealed a defect of Elk-1 activation in the patient cells. These data identify a novel postreceptor defect of insulin and growth factors involving activation of transcription. PMID- 10679247 TI - Edg-6 as a putative sphingosine 1-phosphate receptor coupling to Ca(2+) signaling pathway. AB - The endothelial differentiation gene-6 (Edg-6) was recently identified as an orphan G-protein-coupled receptor. Its predicted amino acid sequence is very close to Edg family of receptor proteins whose ligand is supposed to be lysophosphatidic acid (LPA) or lysosphingolipid such as sphingosine 1-phosphate (S1P) and sphingosylphosphorylcholine (SPC). Transfection of the Edg-6 into Chinese hamster ovary (CHO) cells and K562 cells resulted in the appearance of high-affinity [(3)H]S1P binding activity. Among lipids employed, S1P and, even though less potent, SPC, displaced the [(3)H]S1P binding, but LPA was inactive. In Edg-6-transfected CHO cells, an increase in cytosolic Ca(2+) concentration in response to S1P or SPC was clearly enhanced without change in the LPA-induced action as compared with the vector-transfected cells. The enhancement of the Ca(2+) response was associated with a significant accumulation of inositol phosphate, reflecting activation of phospholipase C. Similar enhancement of Ca(2+) response to S1P or SPC was also observed in Edg-6-expressing K562 cells. These lipid-induced actions in CHO cells and K562 cells expressing Edg-6 were markedly suppressed by pertussis toxin treatment. We conclude that Edg-6 is one of S1P or lysosphingolipid receptors that couple to phospholipase C-Ca(2+) system through pertussis toxin-sensitive G-proteins. PMID- 10679248 TI - The oocyte-specific methylated region of the U2afbp-rs/U2af1-rs1 gene is dispensable for its imprinted methylation. AB - Imprinted genes harbor discrete regions which are differentially methylated in gametes; usually the final differential methylation patterns in adults are established during embryogenesis through modifications of the initial methylation patterns in gametes. Previous reports have shown that a 200-bp region termed region II within the CpG island of the mouse imprinted U2afbp-rs gene is methylated in oocytes but not in sperm, suggesting that this region is a center for the propagation of methylated CpGs on the maternal allele and is also a candidate for an imprinting control element. To determine whether region II is required for the imprinted methylation of this gene at the endogenous locus, we generated mice carrying a deletion of this region. We herein show that parental methylation differences still exist in the CpG island on the region II-deleted allele. These findings suggest that region II is dispensable for the imprinted methylation of the U2afbp-rs gene. PMID- 10679249 TI - Identification of three alternative first exons and an intronic promoter of human PDE5A gene. AB - In the accompanying paper we present evidence for the existence of three PDE5A isoforms that differed only in the 5' end of the mRNAs. In this paper we present evidence that the three isoform-specific 5' ends were encoded by three alternative first exons that were arranged in the order of A1-A3-A2. Because the isoform-specific mRNAs could be transcribed from individual promoters, DNA fragments of the two intronic regions (A1-A3 and A3-A2) were tested for possible promoter activities. The intron between A1- and A3-specific exons did not exhibit any promoter activities even in smooth muscle cells that expressed the A3 isoform (see accompanying paper). In contrast, the intron between A3- and A2-specific exons had promoter activities in PDE5A2-expressing COS-7 and smooth muscle cells. This intronic promoter was bound by transcription factors AP-2 and Sp1, but not by AP-1, as shown by DNase I footprint analysis. However, the sequence bound by AP-2 (5'-GGGAAACGCTCGCGGGAGAGTTGG) is unusual in that it bears little resemblance to the consensus AP-2-binding sequence. PMID- 10679250 TI - Sphingomyelin potentiates chemotherapy of human cancer xenografts. AB - We propose that one manifestation of altered sphingolipid metabolism within tumor cells may be a reduced sensitivity to anti-cancer therapies because of an inability to produce a sufficient apoptotic signal via sphingomyelin hydrolysis to ceramide. If so, then sphingomyelin administration could reverse this effect and increase a tumor's sensitivity to chemotherapy. In vivo, intravenous sphingomyelin (10 mg/day, 7 days) potentiated 5-fluorouracil chemotherapy (0.45 mg/day, 5 days) when co-administered to HT29 human colonic xenograft-bearing nude mice. In vitro, sphingomyelin (SM) at its maximum tolerated concentration increased 5-fluorouracil and doxorubicin sensitivity of HCT15 and MOSER (1 mg/ml SM) and LS174T and SW480 human colonic tumor cells (0.1 mg/ml) approximately 100 300%. At 1 mg/ml SM, however, no effect was seen using HT29, LoVo and WiDr cells. There was no sensitization of normal human umbilical cord endothelial cells. Thus, sphingomyelin co-administration may be one method to improve the selective efficacy of chemotherapy in some tumors, possibly through enhancement of the apoptotic response. PMID- 10679251 TI - Transforming growth factors beta(1) (TGF-beta(1)) and TGF-beta(2) promote glioma cell migration via Up-regulation of alpha(V)beta(3) integrin expression. AB - The migratory behaviour of malignant gliomas relies on the interaction of integrins with extracellular matrix (ECM) components. Transforming growth factor beta(1) (TGF-beta(1)) potently stimulates glioma cell motility whereas TGF beta(2) is known for its immunosuppressive properties. Here, we show that both TGF-beta(1) and TGF-beta(2) promote migration of glioma cells. In parallel, TGF beta(1) and TGF-beta(2) induce alpha(V) and beta(3) intergrin mRNA expression and enhance cell surface expression of alpha(V)beta(3) integrin. TGF-beta-mediated promotion of migration is abrogated by echistatin, a Arg-Gly-Asp (RGD) peptide antagonist of alpha(V)beta(3) integrin, and by a neutralizing anti alpha(V)beta(3) integrin antibody. Taken together, we report a novel mechanism by which TGF-beta modulates cell ECM interactions and promotes glioma cell motility. PMID- 10679252 TI - Missense variations of the gene responsible for Wolfram syndrome (WFS1/wolframin) in Japanese: possible contribution of the Arg456His mutation to type 1 diabetes as a nonautoimmune genetic basis. AB - Recently, a novel gene for a putative transmembrane protein (WFS1/wolframin) was found to be mutated in patients with Wolfram syndrome or DI-DM-OA-D (diabetes insipidus, diabetes mellitus, optic atrophy, and deafness) syndrome. It is suggested that the WFS1 protein is important in the survival of islet beta-cells. We studied the WFS1 gene in a Japanese population to assess its possible role in common type 1 diabetes. Mutation screening revealed four missense mutations; R456H, G576S, H611R, and I720V. By genetic association studies of 185 type 1 diabetes patients and 380 control subjects, we found that R456H was significantly increased in the type 1 diabetes group compared to the control group (P = 0.0005); H611R and I720V were also significantly increased with weaker significance. Furthermore, in patients with the R456H mutation, type 1 diabetes resistant HLA-DRB1 alleles (DRB1*0406, 1501, and 1502) were significantly increased compared to mutation-negative patients while susceptible DRB1*0901 was significantly decreased. Frequencies of autoimmunity characteristics (ICA or GAD Ab positiveness and combination of autoimmune thyroid disease) were decreased in the R456H-positive patients compared to the R456H-negative patients. These data suggest that the WFS1 gene may have a role in the development of common type 1 diabetes as a nonautoimmune genetic basis. PMID- 10679253 TI - Cloning and characterization of mRNA capping enzyme and mRNA (Guanine-7-) methyltransferase cDNAs from Xenopus laevis. AB - The mRNA cap structure, which is synthesized by a series of reactions catalyzed by capping enzyme, mRNA (guanine-7-)-methyltransferase, and mRNA (ribose-2'-O-) methyltransferase, has crucial roles for RNA processing and translation. Methylation of the cap structure is also implicated in polyadenylation-mediated translational activation during Xenopus oocyte maturation. Here we isolated two Xenopus laevis cDNAs, xCAP1a and xCAP1b, for mRNA capping enzyme and one cDNA for mRNA (guanine-7-)-methyltransferase, xCMT1, which encode 598, 511, and 402 amino acids, respectively. The deduced amino acid sequence of xCAP1a was highly homologous to that of human capping enzyme hCAP1a, having all the characteristic regions including N-terminal RNA 5'-triphosphatase as well as C-terminal mRNA guanylyltransferase domains which are conserved among animal mRNA guanylyltransferases, whereas in xCAP1b the most C-terminal motif was missing. The amino acid sequence of xCMT1 was also similar to human (guanine-7-) methyltransferase, hCMT1a, with all the conserved motifs among cellular (guanine 7-)-methyltransferases, except for its N-terminal portion. The recombinant xCAP1a and xCMT1 exhibited cap formation and mRNA (guanine-7-)-methyltransferase activities, respectively. RT-PCR analysis showed that mRNA for xCAP1a and xCMT1 exist abundantly in fertilized eggs as maternal mRNAs, but xCMT1 mRNA gradually decreased in its amount in later stages of early development. PMID- 10679254 TI - ATP synthesis in rod outer segments of bovine retina by the reversal of the disk Ca(2+) pump. AB - Purified disk membranes from rod outer segments of the bovine retina were able to synthesize ATP with a maximal activity (about 52 nmoles ATP/min/mg of protein) at physiological calcium concentrations. This activity was inhibited by vanadate or thapsigargin but not by oligomycin, suggesting the reversal functioning of the disk Ca(2+)-ATPase, which would act as a ATP synthesizer at the expense of the calcium gradient between the disks and the cytoplasm of the rod outer segment. The results are discussed in terms of the need of an immediate source of ATP on the disk membranes where the energy is required to supply the rapid reactions of the photoreception processes. PMID- 10679255 TI - Expression of three isoforms of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in human penile cavernosum. AB - Inhibition of cGMP-specific phosphodiesterase type V (PDE5) has been shown to improve penile erection in patients with erectile dysfunction. We report here the cloning of three PDE5 isoforms from human penile tissues. Two of the isoforms were identical to PDE5A1 and PDE5A2, respectively, which had been isolated from nonpenile tissues. The third isoform was novel and hence called PDE5A3. The deduced amino acid sequence of PDE5A3 was the same as the C-terminal 823-residue sequence of PDE5A1 and PDE5A2. While PDE5A1 and A2 isoforms were expressed in all tissues examined, the A3 isoform was confined to tissues with a smooth muscle or cardiac muscle component. When expressed in COS-7 cells, PDE5A1, A2, and A3 isoforms had similar cGMP-catalytic activities with K(m) of 6.2, 5.75, and 6.06 microM, respectively. Their cGMP-catalytic activities were inhibited by zaprinast with IC(50) values of 3.2 microM, 1.3 microM, and 1.6 microM, respectively, and by sildenafil with IC(50) of 28, 14, and 13 nM, respectively. PMID- 10679256 TI - C2-Ceramide increases cytoplasmic calcium concentrations in human parathyroid cells. AB - Effects of extracellular calcium ([Ca(2+)](ext)) on parathyroid cells are mainly due to the activation of a plasma membrane calcium receptor (CaR) coupled with release of intracellular calcium. In addition, high [Ca(2+)](ext) activates the sphingomyelin pathway in bovine parathyroid cells, generating ceramides and sphingosine. This study explored the direct effects of synthetic ceramides on [Ca(2+)](i) in human parathyroid cells. Cells from five parathyroid adenomas removed from patients with primary hyperparathyroidism were dispersed and maintained in primary culture. Intracellular calcium concentration ([Ca(2+)](i)) [Ca(2+)](i) was monitored using standard quantitative fluorescence microscopy in Fura-2/AM-loaded cells. Laser scanning microscopy was used to monitor the intracellular distribution of a fluorescent ceramide analogue (BODIPY-C5). After addition of 10 microM C2-ceramide (N-acetyl-d-erythro-sphingosine), [Ca(2+)](i) increased rapidly (30-60 s) to a peak three times above basal levels in 70% of cells (37/55 cells in four experiments). This effect appeared to be due to release of Ca(2+) from intracellular stores rather than Ca(2+) entry from the extracellular medium. C2-responsive cells had a smaller [Ca(2+)](i) response to subsequent stimulation with the CaR agonist-neomycin (1 mM). These responses were specific to C2 since C6-ceramide (N-hexanoyl-d-erythro-sphingosine) did not affect basal [Ca(2+)](i) nor the responses to an increase in [Ca(2+)](ext) and to neomycin. C5-BODIPY generated intense perinuclear fluorescence, suggesting targeting of the ceramides to the Golgi apparatus. These data demonstrate that endogenous generation of ceramides has the potential to modulate changes in [Ca(2+)](i) and secretion in response to [Ca(2+)](ext) in human parathyroid cells. PMID- 10679257 TI - Oxidative stress induces increase in intracellular amyloid beta-protein production and selective activation of betaI and betaII PKCs in NT2 cells. AB - Amyloid beta-protein (Abeta) aggregation produces an oxidative stress in neuronal cells that, in turn, may induce an amyloidogenic shift of neuronal metabolism. To investigate this hypothesis, we analyzed intra- and extracellular Abeta content in NT2 differentiated cells incubated with 4-hydroxy-2,3-nonenal (HNE), a major product of lipid peroxidation. In parallel, we evaluated protein kinase C (PKC) isoenzymes activity, a signaling system suspected to modulate amyloid precursor protein (APP) processing. Low HNE concentrations (0.1-1 microM) induced a 2-6 fold increase of intracellular Abeta production that was concomitant with selective activation of betaI and betaII PKC isoforms, without affecting either cell viability or APP full-length expression. Selective activation of the same PKC isoforms was observed following NT2 differentiation. Our findings suggest that PKC beta isoenzymes are part of cellular mechanisms that regulate production of the intracellular Abeta pool. Moreover, they indicate that lipid peroxidation fosters intracellular Abeta accumulation, creating a vicious neurodegenerative loop. PMID- 10679258 TI - Inhibition of MAP kinase kinase (MEK) results in an anti-inflammatory response in vivo. AB - The MAP kinase pathway has been well-characterized as a cascade of sequential protein phosphorylation events leading to the upregulation of a variety of genes in response to growth factors and mitogens. We are interested in the role of these kinases in inflammation and have thus examined their activity in vivo using TPA-induced ear edema in the mouse as a model of inflammation. We show that the activities of both ERK-1 and ERK-2 are upregulated in this model in response to TPA. Increased levels of ERK phosphorylation are measurable as early as 15 min poststimulation and reach a level 8-fold over controls at 4 h. In contrast, minimal activation of JNK or p38 is observed. Topical treatment of ears with the MEK inhibitor, U0126, prevents ERK phosphorylation and ear swelling in a dose dependent manner in this model. These results suggest that the MEK/ERK pathway is important during an inflammatory response in vivo. PMID- 10679259 TI - Zinc induces the accumulation of hypoxia-inducible factor (HIF)-1alpha, but inhibits the nuclear translocation of HIF-1beta, causing HIF-1 inactivation. AB - The replacement of heme iron by cobalt or nickel in a putative oxygen sensor is supposed to reduce oxygen binding to the heme protein, resulting in HIF-1 activation and erythropoietin (EPO) induction. According to this hypothesis, zinc might be another example of a transition metal which is capable of stimulating EPO production. By substituting for heme iron, zinc protoporphyrin IX is produced, which has a known low oxygen affinity. However, it has been reported that zinc fails to induce EPO in normoxia, and that it suppresses EPO production in hypoxic cells. This unexpected effect of zinc on EPO production is not understood. In this study, we found that zinc induced the accumulation and nuclear translocation of hypoxia-inducible factor (HIF)-1alpha but inhibited the nuclear translocation of HIF-1beta, which inactivated HIF-1 and suppressed EPO mRNA induction in hypoxic cells. PMID- 10679260 TI - Lipopolysaccharide augments expression and secretion of vascular endothelial growth factor in rat ventricular myocytes. AB - Vascular endothelial growth factor (VEGF), also known as vascular permeability factor, is highly expressed in the myocardium under various stimuli including hypoxia and ischemia. On the other hand, lipopolysaccharide (LPS) causes systemic inflammatory response syndrome (SIRS), which consists of systemic pathophysiological changes related to vascular hyperpermeability. To test the hypothesis that VEGF is one of the important mediators of SIRS, we examined effects of LPS on the VEGF expression and secretion in cultured neonatal rat ventricular myocytes. LPS (10 microg/ml) rapidly (within 1 h) augmented the levels of VEGF mRNA in these cells. Pharmacological inhibition of nucleic factor kappaB or tyrosine kinases did not affect the LPS-induced augmentation of VEGF mRNA expression, while these treatments markedly suppressed the up-regulation of inducible nitric oxide synthase (iNOS) expression by LPS. The VEGF concentrations in the conditioned media were also significantly increased by the LPS treatment of 6 h. In conclusion, LPS augments VEGF expression and secretion in rat ventricular myocytes, suggesting that VEGF may be involved in pathogenesis of SIRS. LPS may induce VEGF mRNA through the signaling pathways that are distinct from those responsible for the iNOS induction. PMID- 10679261 TI - Heme metabolism of Plasmodium is a major antimalarial target. AB - The malarial parasite manifests unique features of heme metabolism. In the intraerythrocyte stage it utilizes the host hemoglobin to generate amino acids for its own protein synthesis, but polymerizes the acquired heme as a mechanism for detoxification. At the same time the parasite synthesizes heme de novo for metabolic use. The heme biosynthetic pathway of the parasite is similar to that of hepatocytes and erythrocytes. However, while the parasite makes its own delta aminolevulinate (ALA) synthase that is immunochemically different from that of the host, it imports ALA dehydrase and perhaps the subsequent enzymes of the pathway from the host red cell. Many schizonticidal drugs such as chloroquine and artemisinin act by interfering with the heme metabolism of the parasite and there is scope to design new molecules based on the unique features of this metabolic machinery in the parasite. PMID- 10679262 TI - Age-related decline in osteoprotegerin expression by human bone marrow cells cultured in three-dimensional collagen sponges. AB - With advancing age, an increase in bone resorption relative to bone formation results in bone loss. Bone marrow stromal cells and their products support osteoclastogenesis from hematopoietic progenitors. Another of their products, osteoprotegerin (OPG), blocks the osteoclast-stimulatory effects of OPG ligand. We tested the hypothesis that with advancing age there is a decrease in OPG expression by human bone marrow cells. Bone marrow cells were obtained from 18 subjects (age range 38-84 years). Expression of mRNA transcripts of OPG was assessed by quantitative competitive RT-PCR. Median number of OPG transcripts in the younger group was 0. 3 zetptomoles (range 0.01 to 1.30) and was higher than in the older group's median of 0.06 (range 0 to 0.5; p < 0.05). The decline in the expression of OPG with age may increase the capacity of stromal/osteoblast cells to support osteoclastogenesis. PMID- 10679263 TI - Dramatic magnesium efflux induced by high potassium in rat thymocytes. AB - When incubated in 150 mM KCl, rat thymocytes exhibited a very important magnesium efflux (11.4 +/- 0.7 mmoles/liter cells/20 min, n = 29), about 90 times higher than the physiological magnesium efflux catalyzed by the Na-Mg exchanger (0.126 +/- 0.093 mmoles/liter cells/20 min). Cells remained viable (trypan blue test) and membrane integrity was shown by the absence of an increase in sodium permeability. K(+)-induced magnesium efflux exhibited the following properties: (i) it required the presence of external chloride; (ii) it was fully blocked by DIOA, a selective KCl-cotransporter inhibitor (IC(50) = 35 microm); and (iii) it was associated to a progressive increase in cell volume via the DIOA-sensitive K Cl cotransporter. Such cell swelling seems to play a causal role, because (i) hypertonic media (+400 mM sucrose) abolished K(+)-induced magnesium efflux and (ii) hypotonic Ringer media (205 mOsm) increased both cell volume and magnesium efflux (from a basal value of 0.35 +/- 0.03 mmoles/liter cells/20 min up to 1.44 +/- 0.24 mmoles/liter cells/20 min), even in the presence of DIOA. In conclusion, high potassium induced a dramatic release of intracellular magnesium from rat thymocytes. Such a phenomenon was, at least in part, caused by cell swelling via the DIOA-sensitive K-Cl cotransporter. The nature of the magnesium transport mechanism and its role in the transduction signal of K-Cl cotransporter activation by cell swelling deserve further investigation. PMID- 10679264 TI - Cloning and expression of cytochrome P450 genes belonging to the CYP4 family and to a novel family, CYP48, in two hymenopteran insects, Trichogramma cacoeciae and Apis mellifera. AB - Cytochrome P450 partial sequences were isolated by PCR using genomic DNA from two hymenopteran insects of agronomical importance, Trichogramma cacoeciae, a parasitoid wasp, and Apis mellifera, the honeybee. Four new P450 genes were identified: one honeybee gene belongs to the CYP4 family and was named CYP4G11; the three other genes were from Trichogramma and belong to the CYP4 family (CYP4G12) and to a novel family, the CYP48 one (CYP48A1 and CYP48A2). The four genes contain a short intron (72-95 bp) at the same position as already described for other insect species. The two genes CYP48A1 and CYP48A2 have a supernumary intron (57-71 bp) upstream the first one. Only the two CYP4 genes were constitutively transcribed, at a high level for CYP4G12 and at a low level for CYP4G11. No expression was observed for CYP48A1 and CYP48A2. PMID- 10679265 TI - The effect of water on the Fe(3+)/Fe(2+) reduction potential of heme. AB - Hemeproteins can act as catalysts, oxygen carriers or electron conductors. The ferric/ferrous reduction potential E(m7) of iron in the center of the prosthetic group ranges from negative values for peroxidases to an extreme positive value for cytochrome a(3) with Hb and Mb in the middle [1]. Proteins exercise their influence on E(m7) in several ways: via substituents at the periphery of the chelate structure, via the proximal ligand, and via interaction with the surrounding medium, amino acid side chains, or polar solvents. Work on recombined proteins and 2,4-substituted free hemes documented that the first two effects are additive [2]. For the third effect, models of the dielectric media on a molecular level have been successfully applied [3-5]. E(m7) has also been empirically correlated to the degree of heme exposure to water [6-8]. The apoprotein/porphyrin and water/porphyrin interfaces are complementary since water molecules fill any empty space in the crevice and surround any pertinent part of heme outside the protein boundary. The present work links to this idea by a combination of statistical mechanics simulations and quantum mechanical calculations comparing heme in water with heme in an apolar environment. Our results show that polarization of the porphyrin pi-electron cloud by the field from water dipoles influences E(m7). The dominant effect of this and other determinates of iron electron availability is perturbations of delocalized electron density in the porphyrin chelate, reproduced by a model where the prosthetic group is treated as a disc of uniform electron density. The present work is also of interest since the interfacial energy constitutes the main barrier for heme-protein separation [9-11]. PMID- 10679266 TI - Expression of a Desulfovibrio tetraheme cytochrome c in Escherichia coli. AB - A tetraheme cytochrome c was successfully overexpressed for the first time in Escherichia coli. Desulfovibrio desulfuricans ATCC 27774 tetraheme cytochrome c(3) was expressed in aerobically grown Escherichia coli cotransformed with Escherichia coli ccm gene cluster (Arslan et al. (1998) Bioch. Biophys. Res. Commun. 251, 744-747). The analysis of the produced cytochrome showed that the signal peptide was correctly cleaved, the four heme groups were inserted and the electronic structure around the heme irons was conserved, i.e., the recombinant tetraheme cytochrome was identical to that isolated from the native source. Contradicting previous results which indicated that Escherichia coli was only capable of producing apocytochrome c(3) (Pollock et al. (1989) J. Gen. Microbiol. 135, 2319-2328), the present work proves unequivocally that the holoform can also be obtained. PMID- 10679267 TI - Human glial cell-line-derived neurotrophic factor: a structure-function analysis. AB - Glial cell-line-derived neurotrophic factor (GDNF) is a protein known to enhance the survival of dopaminergic and motor neurons. It has been shown to have therapeutic potential in the treatment of Parkinson's disease and other neurodegenerative diseases. GDNF gene was modified by deletion and insertion mutagenesis using PCR methods. The various mutants were all highly expressed in Escherichia coli. The recombinant proteins were purified and their survival promoting activities were determined by motor neurons. The result showed that the C-terminus was critical for structure stability of GDNF, and the alpha-helix, finger1 and finger2 regions were involved in receptor binding, while the N terminus was not essential for the biological functions of GDNF. PMID- 10679268 TI - Molecular cloning of murine STAP-1, the stem-cell-specific adaptor protein containing PH and SH2 domains. AB - To identify the novel substrate of c-kit which is important for hematopoietic stem cell self-renewal or differentiation, CD34-low/negative, Sca-1-positive, c kit-positive, and lineage marker-negative (CD34(low/-)Sca-1(+)c-kit(+)Lin(-)) cells were sorted by a fluorescence-activated cell sorter from mouse bone marrow cells and a yeast two-hybrid cDNA library was constructed. By screening with c kit as bait, we cloned a novel cDNA, designed STAP-1, encoding an adaptor protein with a Pleckstrin homology domain, the Src homology 2 (SH2) domain, and a number of tyrosine phosphorylation sites. RT-PCR analysis revealed that STAP-1 expression is restricted in the bone marrow cell fraction expressing c-kit. The highest expression was observed in the CD34(low/-)Sca-1(+)c-kit(+)Lin(-) stem cell-enriched fraction. The murine myeloid cell line, M1, expressed a high level of STAP-1. However, the expression was strongly repressed in response to leukemia inhibitory factor (LIF) which induced monocytic differentiation of M1 cells, suggesting that STAP-1 is associated with the undifferentiated cell type. A two hybrid assay indicated that STAP-1 bound not only to c-kit but also to c-fms but not to JAK2 or Pyk2. In 293 cells, STAP-1 was tyrosine-phosphorylated by activated c-kit. An in vitro binding assay suggested that the STAP-1 SH2 domain interacted with several tyrosine-phosphorylated proteins including c-kit and STAT5. These suggest that STAP-1 functions as an adaptor molecule downstream of c kit in hematopoietic stem cells. PMID- 10679269 TI - The human PEX3 gene encoding a peroxisomal assembly protein: genomic organization, positional mapping, and mutation analysis in candidate phenotypes. AB - In yeasts, the peroxin Pex3p was identified as a peroxisomal integral membrane protein that presumably plays a role in the early steps of peroxisomal assembly. In humans, defects of peroxins cause peroxisomal biogenesis disorders such as Zellweger syndrome. We previously reported data on the human PEX3 cDNA and its protein, which in addition to the peroxisomal targeting sequence contains a putative endoplasmic reticulum targeting signal. Here we report the genomic organization, sequencing of the putative promoter region, chromosomal localization, and physical mapping of the human PEX3 gene. The gene is composed of 12 exons and 11 introns spanning a region of approximately 40 kb. The highly conserved putative promoter region is very GC rich, lacks typical TATA and CCAAT boxes, and contains potential Sp1, AP1, and AP2 binding sites. The gene was localized to chromosome 6q23-24 and D6S279 was identified to be the closest positional marker. As yeast mutants deficient in PEX3 have been shown to lack peroxisomes as well as any peroxisomal remnant structures, human PEX3 is a candidate gene for peroxisomal assembly disorders. Mutation analysis of the human PEX3 gene was therefore performed in fibroblasts from patients suffering from peroxisome biogenesis disorders. Complementation groups 1, 4, 7, 8, and 9 according to the numbering system of Kennedy Krieger Institute were analyzed but no difference to the wild-type sequence was detected. PEX3 mutations were therefore excluded as the molecular basis of the peroxisomal defect in these complementation groups. PMID- 10679270 TI - Apoaequorin monitors degradation of endoplasmic reticulum (ER) proteins initiated by loss of ER Ca(2+). AB - Apoaequorin was targeted to the cytosol, nucleus, and endoplasmic reticulum of HeLa cells in order to determine the effect of Ca(2+) release from the ER on protein degradation. In resting cells apoaequorin had a rapid half-life (ca. 20 30 min) in the cytosol or nucleus, but was relatively stable for up to 24 h in the ER (t(1/2) > 24 h). However, release of Ca(2+) from the ER, initiated by the addition of inhibitors of the ER Ca(2+)/Mg(2+) ATPase such as 2 microM thapsigargin or 1 microM ionomycin, initiated rapid loss of apoaequorin in the ER, but had no detectable effect on apoaequorin turnover in the cytosol nor the nucleus. This loss of apoprotein was not the result of secretion into the external fluid, and could not be inhibited by inhibitors of protein degradation by proteosomes. Proteolysis of apoaequorin in cell extracts (t(1/2) < 20 min) was completely inhibited in the presence of 1 mM Ca(2+), and this effect was independent of the ER retention signal KDEL at the C-terminus. Proteolysis was unaffected by the presence of selected serine protease inhibitors, or 10 microM Zn(2+), a known caspase-3 inhibitor. The results show that apoaequorin can monitor proteolysis of ER proteins activated by loss of ER Ca(2+). Several Ca(2+) binding proteins exist in the ER, acting as the Ca(2+) store and chaperones. Our results have important implications both for the role of ER Ca(2+) in cell activation and stress and when using aequorin for monitoring free ER Ca(2+) over long time periods. PMID- 10679271 TI - Fungal gliotoxin targets the onset of superoxide-generating NADPH oxidase of human neutrophils. AB - Gliotoxin from Aspergillus, bearing a S&bond;S bond in its structure, prevented the onset of O(-)(2) generation by the human neutrophil NADPH oxidase in response to phorbol myristate acetate (PMA). Gliotoxin affected the activation process harder than the activated oxidase, as shown by its stronger inhibition when added to neutrophils prior to, than post-PMA at maximum enzyme turnover. Decreased O( )(2) generation persisted even if cells treated with gliotoxin were subsequently washed, with half-inhibition concentrations (IC(50)) of 5.3, and 3.5 microM for treatments of 15 and 30 min, respectively. In addition, gliotoxin made neutrophils reduce cytochrome c regardless of absence of PMA, through its reaction with intracellular reductants in an oxygen-dependent process, named redox cycling. Thus, we next tested whether preincubation of neutrophils with gliotoxin under hypoxic conditions would relieve the inhibition of NADPH oxidase. Instead, this prevention of redox cycling significantly favored damage to the NADPH oxidase with an IC(50) of 0.009 microM. Moreover, conversion of gliotoxin to its dithiol derivative by addition of reduced dithiothreitol during incubation protected cells from losing oxidase activity. These findings support that the disulfide form of gliotoxin targets NADPH oxidase activation. PMID- 10679272 TI - 3-Methyladenine-DNA glycosylase I from Escherichia coli-computer modeling and supporting experimental evidence. AB - TagA (3-methyladenine-DNA glycosylase I) excises 3-methyadenine and 3 methylguanine from alkylated DNA. The structure of this enzyme has not yet been determined experimentally. We propose a three-dimensional model of the TagA protein based on the threading algorithm. The model shows that TagA is a mostly alpha-helical protein, in agreement with circular dichroism measurements. None of the eight cysteines present in the TagA sequence forms a disulfide bridge in the model structure, which has also been experimentally verified with the use of Ellman method. PMID- 10679273 TI - In situ demonstration of inhibitory effects of hammerhead ribozymes that are targeted to the hepatitis Bx sequence in cultured cells. AB - Chronic hepatitis B virus (HBV) infection is endemic to several populous areas of the world and is frequently complicated by hepatocellular carcinoma. Ribozymes can be designed to cleave target RNA sequences specifically and show promise for the treatment of HBV infection. Demonstration of intracellular inhibition of HBV gene expression, essential to developing therapeutic ribozymes, has been the aim of this investigation. We generated two vectors encoding hammerhead ribozymes that target the HBx region of HBV. Plasmids containing intact HBV sequences or a modification in which the preS2/S region was replaced by DNA encoding enhanced green fluorescent protein (EGFP) were used to test ribozyme action in transfected cells. Both ribozymes inhibited surface antigen secretion and EGFP expression similarly. The measurement of EGFP expression is convenient to assess ribozyme action in situ and effective targeting of HBV sequences that are common to all HBV transcripts is potentially useful to develop strategies to counter HBV infection. PMID- 10679274 TI - Receptor-mediated gene delivery approach demonstrates the role of 5'-proximal DNA region in conferring phenobarbitone responsiveness to CYP2B2 gene in rat liver in vivo. AB - The phenobarbitone (PB) responsiveness of the 5'-proximal region of the CYP2B1/B2 gene was examined in detail with plasmid DNA constructs containing G-free cassette as reporter, using in vivo targeting of the same DNA constructs into rat liver as galactosylated-polylysine complexes. The contribution of the proximal region (-1 to -179 bp) and the positive element (-69 to -98 bp) identified earlier in this laboratory to PB responsiveness was assessed. The results obtained on PB treatment of rats subjected to receptor-mediated gene delivery to liver were conclusive and dramatic, with the control (saline-treated) rats manifesting very little expression of the reporter, reflecting the in vivo picture of CYP2B1/B2 gene expression. The positive element conferred PB responsiveness to homologous and heterologous promoters. Deletion of the positive element led to elimination of PB response. The entire -179 bp region was significantly more effective in responding to PB treatment than the region up to 98 bp, both containing one copy of the positive element. Thus, the positive element and its flanking sequences in the 5'-proximal region are involved in conferring PB responsiveness to the CYP2B1/B2 gene. PMID- 10679275 TI - Clay-bridged electron transfer between cytochrome p450(cam) and electrode. AB - We demonstrate a very fast heterogeneous redox reaction of substrate-free cytochrome P450(cam) on a glassy carbon electrode modified with sodium montmorillonite. The linear relationship of the peak current in the cyclic voltammogram with the scan rate indicates a reversible one-electron transfer surface process. The electron transfer rate is in the range from 5 to 152 s(-1) with scan rates from 0.4 to 12 V/s, respectively. These values are comparable to rates reported for the natural electron transfer from putidaredoxin to P450(cam). The formal potential of adsorbed P450(cam) is -139 mV (vs NHE) and therefore positively shifted by 164 mV compared to the potential of substrate-free P450(cam) in solution. UV-VIS and FTIR spectra do not indicate an influence of the clay colloidal particles on the heme and the secondary structure of P450(cam) in solution. However, P450(cam) adsorbed on the surface of the clay-modified electrode may undergo partial dehydration resulting in the shift of the formal potential. PMID- 10679276 TI - Biochemical/spectroscopic characterization and preliminary X-ray analysis of a new aldehyde oxidoreductase isolated from Desulfovibrio desulfuricans ATCC 27774. AB - Aldehyde oxidoreductase (AOR) activity has been found in different sulfate reducing organisms (Moura, J. J. G., and Barata, B. A. S. (1994) in Methods in Enzymology (Peck, H. D., Jr., and LeGall, J., Eds.), Vol. 243, Chap. 4. Academic Press; Romao, M. J., Knablein, J., Huber, R., and Moura, J. J. G. (1997) Prog. Biophys. Mol. Biol. 68, 121-144). The enzyme was purified to homogeneity from extracts of Desulfovibrio desulfuricans (Dd) ATCC 27774, a sulfate reducer that can use sulfate or nitrate as terminal respiratory substrates. The protein (AORDd) is described as a homodimer (monomer, circa 100 kDa), contains a Mo-MCD pterin, 2 x [2Fe-2S] clusters, and lacks a flavin group. Visible and EPR spectroscopies indicate a close similarity with the AOR purified from Desulfovibrio gigas (Dg) (Barata, B. A. S., LeGall, J., and Moura, J. J. G. (1993) Biochemistry 32, 11559-11568). Activity and substrate specificity for different aldehydes were determined. EPR studies were performed in native and reduced states of the enzyme and after treatment with ethylene glycol and dithiothreitol. The AORDd was crystallized using ammonium sulfate as precipitant and the crystals belong to the space group P6(1)22, with unit cell dimensions a = b = 156.4 and c = 177.1 A. These crystals diffract to beyond 2.5 A resolution and a full data set was measured on a rotating anode generator. The data were used to solve the structure by Patterson Search methods, using the model of AORDg. PMID- 10679277 TI - Amyloid-beta peptides interact with plasma proteins and erythrocytes: implications for their quantitation in plasma. AB - Amyloid beta peptides are bound rapidly in the plasma complicating an accurate assessment of their in vivo abundance by immunoassay procedures. The extent of Abeta immunoassay interference was used to estimate the Abeta binding capacity of purified plasma proteins, erythrocytes and whole plasma. Human serum albumin bound Abeta peptides rapidly with a 1:1 stoichiometry and at physiological concentrations was capable of binding over 95% of an input of 5 ng/ml Abeta. Purified alpha2-macroglobulin was able to bind Abeta peptides and at physiological concentration bound 73% of 5 ng/ml of Abeta. Erythrocytes also sequestered the Abeta peptides, showing a preference for binding Abeta 1-42. Incubation of 5 ng/ml of Abeta in plasma revealed that about 30% of the peptides were still detectable by immunoassay, presumably reflecting the binding of Abeta peptides with albumin and other plasma molecules. Thus, our studies reveal that both the soluble and formed elements of the blood are capable of sequestering Abeta peptides. To avoid underestimating plasma Abeta values, we employed an improved column chromatography method under denaturing conditions to liberate Abeta from its associations with plasma proteins. Quantification of Abeta 40 and 42 levels in plasma from both normal and AD individuals after chromatography showed a large overlap between AD and control groups, despite the very large pool of Abeta present in the AD brains. The potential origins of the plasma Abeta pool are discussed. PMID- 10679278 TI - Involvement of ERK in BMP-2 induced osteoblastic differentiation of mesenchymal progenitor cell line C3H10T1/2. AB - The signaling mechanisms responsible for bone morphogenetic protein (BMP) induced osteoblast differentiation remains poorly understood. Previous research demonstrated that Smad proteins are the substrates and the mediators of BMP bound serine/threonine receptor kinase. In the present study, we examined the possible involvement of extracellular signal-regulated kinase (Erk) in the BMP induced osteoblast differentiation of mesenchymal progenitor cell C3H10T1/2. Our results indicate that BMP-2 inducement increased MAP kinase activity in mesenchymal progenitor cell line C3H10T1/2. Contrary to previous reports, this increased MAP kinase activity showed a latent but sustained pattern. Elevation of Erk1 and Erk2 protein levels was observed simultaneously. RT-PCR results demonstrated that the elevation of Erk protein level in BMP-2 induced cells was from the upregulation of mRNA expression. Furthermore, upregulated Erk proteins present enhanced phosphorylation. By using a dominant-negative Erk2 cell line, we demonstrated that nonfunctional Erk2 partially eliminated BMP-2 induced cell proliferation and ALP activity in the C3H10T1/2 cell. These results indicate that Erk is involved in BMP-2 induced osteoblast differentiation. The results also demonstrate that a latent and sustained signaling pattern exists in BMP induced signaling cascade. PMID- 10679279 TI - Enhanced expression of cellular prion protein gene by insulin or nerve growth factor in immortalized mouse neuronal precursor cell lines. AB - In order to understand the fundamental and putative roles of PrP(c) in the central nervous system, neuronal cell lines were established. Cells were immortalized by recombinant retrovirus vector-mediated transduction of SV40 T antigen gene. Among these, two cell lines were selected based on their RT-PCR expressions of neuron-specific neurofilament (NF-H, NF-M) and cell morphology. These cell lines showed the properties of neuronal progenitor cells in antigenicity, morphology and responses to differentiating agents. Expression of PrP(c) was detected by immunocytochemical analysis. These cell lines responded to differentiating agents such as dibutyl cyclic AMP (dcAMP) and phorbol 12 myristate 13-acetate (PMA) before developing into neuronal-like cells. Neurite extensions were observed 20 min after incubation with the differentiating agents. Treatment with nerve growth factor (NGF) and insulin induced cell differentiation and enhanced expression of PrP gene (Prnp) mRNA and protein. The latter phenomenon was not inhibited by wortmannin, which is a specific inhibitor of phosphatidylinositol 3-kinase. These results suggest that PrP(c) plays an important role in the differentiation-mediated classic signaling pathway of neuronal cell. PMID- 10679280 TI - Green tea polyphenols: novel and potent inhibitors of squalene epoxidase. AB - The green tea gallocatechins, (-)-epigallocatechin-3-O-gallate (EGCG) (IC(50) = 0.69 microM), (-)-gallocatechin-3-O-gallate (GCG) (IC(50) = 0.67 microM), (-) epicatechin-3-O-gallate (ECG) (IC(50) = 1.3 microM), and theasinensin A (IC(50) = 0.13 microM), were found to be potent and selective inhibitors of rat squalene epoxidase (SE), a rate-limiting enzyme of cholesterol biogenesis. On the other hand, flavan-3-ols without galloyl group at C-3 did not show significant enzyme inhibition. It was demonstrated for the first time that the cholesterol lowering effect of green tea may be attributed to their potent SE inhibition activities. Inhibition kinetics revealed that EGCG inhibited SE in noncompetitive (K(I) = 0.74 microM), and non-time-dependent manner. The potent enzyme inhibition would be caused by specific binding to the enzyme, and by scavenging reactive oxygen species required for the monooxygenase reaction. PMID- 10679281 TI - Selective activation of antitumor activity of macrophages by the delivery of muramyl dipeptide using a novel polynucleotide-based carrier recognized by scavenger receptors. AB - We have shown that muramyl dipeptide (MDP) conjugated to a 10-mer polyguanylic acid (PolyG) is specifically internalized by macrophages through scavenger receptor (SCR)-mediated endocytosis. Macrophages activated by PolyG-MDP displayed about 20-fold higher cytotoxic activity against nonmacrophage tumor cells compared to that elicited by free MDP. The PolyG-MDP was found to trigger the secretion of higher levels of interleukin-6, interleukin-1alpha, TNF-alpha, and nitric oxide in comparison to free MDP. Addition of antibodies directed against IL-6 and TNF-alpha to macrophage culture completely abrogated the tumoricidal response of PolyG-MDP, indicating that these two cytokines are primarily responsible for bioefficacy. This general approach of PolyG as a vehicle may find wide application in the delivery of genes and antisense oligonucleotides to macrophages. PMID- 10679282 TI - Expression of 5-lipoxygenase by human colorectal carcinoma Caco-2 cells during butyrate-induced cell differentiation. AB - Butyrate, a short-chain fatty acid, modulates proliferation and differentiation of normal and neoplastic colonocytes. We examined the expression of 5 lipoxygenase (5-LO) and its metabolites in human colorectal carcinoma (Caco-2) cells, exposed to differentiation-inducing doses of butyrate. Treatment with butyrate significantly increased 5-lipoxygenase mRNA and protein in comparison to nontreated cells. Cyclooxygenases (COX)-1 and COX-2 mRNA were not significantly influenced by the treatment. However, 5-LO activity, low in nontreated cells, increased only minimally after butyrate, and its metabolic product (5-HETE) was detectable neither in control nor in treated cells. In contrast, 15-HETE (a product of 15-LO, which is also upregulated by butyrate) rose significantly. We conclude that, although being overexpressed by butyrate on mRNA and protein level, 5-LO remains inactive in differentiating Caco-2 cells. This is likely to be due either to some associated actions of butyrate, or to 5-LO-inhibition by 15 HETE, concomitantly induced by butyrate treatment. PMID- 10679283 TI - Ras-related GTPase RhoB forces alkylation-induced apoptotic cell death. AB - rhoB encoding a Ras-related GTPase is immediate-early inducible by genotoxic treatments. To address the question of the physiological role of RhoB in cellular defense, cells stably overexpressing wild-type RhoB protein were generated. Overexpression of RhoB renders cells hypersensitive to the killing effect of alkylating agents including antineoplastic drugs but not to UV-light and doxorubicin. As compared to control cells, RhoB overexpressing cells revealed an increase in the frequency of alkylation-induced apoptotic cell death. This indicates that RhoB is involved in modulating apoptotic signaling. Furthermore, overexpression of RhoB resulted in a prolonged transient block to DNA replication upon MMS treatment. UV-induced replication blockage was not affected by RhoB. Based on the data we suggest RhoB to be a novel regulatory factor which takes influence on the level of cytotoxicity of DNA damaging drugs and forces cells to alkylation-induced apoptosis. The data indicate that this might be due to RhoB mediated delay in cell cycle progression upon alkylation treatment. PMID- 10679284 TI - Intracellular supply of phospholipids for biliary secretion: evidence for a nonvesicular transport component. AB - Phospholipids (PL) for biliary secretion could be supplied from the endoplasmic reticulum (ER) to the plasma membrane by cytosolic transfer proteins or transport vesicles. Therefore, we studied whether biliary secretions of PL and apolipoprotein A-I (apo A-I), as markers for the ER-to-Golgi vesicular transport pathway, are tightly coupled in isolated perfused rat livers with enhanced secretion (+60%) of PL after withdrawal of the cholesterol synthesis inhibitor pravastatin (0.1% of chow, fed for 7 days). Blocking agents dissociated the secretion of apo A-I and PL. Brefeldin A as well as cycloheximide inhibited biliary secretion of apo A-I (-52%; -68%), however, not of PL. Both bilirubin ditaurate and taurodehydrocholic acid reduced biliary secretion of PL (-27%; 79%), but not of apo A-I. The data support the concept that PL destined for biliary secretion bypass the vesicular transport pathway of apo A-I through the Golgi compartment, most likely via cytosolic transfer proteins. PMID- 10679285 TI - De novo RNA synthesis catalyzed by HCV RNA-dependent RNA polymerase. AB - The 65 kDa RNA-dependent RNA polymerase (NS5B), encoded by the hepatitis C virus (HCV) genome, is a key component involved in viral replication. Here we provide the direct evidence that purified HCV polymerase catalyzed de novo RNA synthesis in a primer-independent manner using homopolymers and HCV RNA as templates. The enzyme could utilize both polyC and polyU as templates for de novo RNA synthesis, suggesting that NS5B specifically recognized pyrimidine bases for initiation. More importantly, NS5B also catalyzed de novo RNA synthesis with an HCV RNA template; the resulting nascent RNA products, smaller than the template used, contained ATP as the first nucleotide. These results indicate that the newly synthesized RNAs did not result from template self-priming and suggest that a replication initiation site in the HCV RNA genome is a uridylate. PMID- 10679286 TI - A two-hit model for development of multiple endocrine neoplasia type 2B by RET mutations. AB - Multiple endocrine neoplasia (MEN) type 2B mutations have been reported at methionine 918 or alanine 883 in the tyrosine kinase domain of the RET proto oncogene. Recently, a new combination of two germline missense mutations at valine 804 and tyrosine 806 was identified in a patient with MEN 2B-like clinical phenotypes including medullary thyroid carcinoma, mucosal neuroma, and marfanoid habitus. In this case, valine 804 and tyrosine 806 were replaced with methionine and cysteine, respectively. In the present study, biological activities of RET with these new mutations were compared with those with known MEN 2A or MEN 2B mutations. The transforming activity of RET with the V804M/Y806C mutation was about 8- to 13-fold higher than that of RET with a single V804M or Y806C mutation. Like RET with the M918T or A883F MEN 2B mutation, the transforming activity of RET with the V804M/Y806C mutation was not affected by substitution of phenylalanine for tyrosine 905 that abolished the activity of RET with the MEN 2A mutation. On the other hand, substitution of phenylalanine for tyrosines 864 and 952 drastically diminished the activity of RET with the V804M/Y806C, M918T or A883F mutation, suggesting that these three mutant proteins have similar biological properties. PMID- 10679287 TI - In vitro remodeling of tumor vascular endothelial cells using conditioned medium from various tumor cells and their sensitivity to TNF-alpha. AB - Prevention of tumor-associated blood vessel formation (angiogenesis) is a potentially powerful strategy to treat cancer. We found that tumor vascular endothelial cells were rearranged in vitro with conditioned culture medium derived from tumor cells and compared the sensitivity to the effects of TNF-alpha between normal and tumor endothelial cells. Incubation with tumor (Meth-A, Colon26)-derived conditioned medium showed that no effect was observed on cell growth. Tumor cells (Meth-A, Colon26, and B16BL6) only showed no sensitivity to TNF-alpha. Normal and control endothelial cells in culture showed little cytotoxicity in response to TNF-alpha treatment, but marked cytotoxicity of TNF alpha was observed in endothelial cells cultured with tumor-derived conditioned medium. Sensitivity to TNF-alpha was different depending on the type of tumor from which the conditioned medium was derived. This difference in sensitivity was assumed to be due to the in vivo sensitivity to TNF-alpha. The results of this study suggested that the sensitivity of tumors to TNF-alpha is controlled by the sensitivity of tumor vasculature. PMID- 10679288 TI - N-Glycan structures of an osteopontin from human bone. AB - N-Glycan structures of osteopontin (a bone matrix protein) from human bone (lumbar vertabrate) are reported in detail. Asn-linked glycan portion was released from 100 microg of osteopontin by digestion with glycoamidase A (from sweet almond), and the reducing ends of the N-glycans were reductively aminated with 2-aminopyridine. The derivatized N-glycans were separated and structurally identified by a multidimensional mapping technique on HPLC columns. Two major N glycan structures were also confirmed by mass spectrometry. The proposed structures are shown below. The result should permit future comparison with the N glycan structures of osteopontins obtained from other sources (kidney tissues, macrophages, urinary stones, human milk, etc.). PMID- 10679289 TI - The kinetics of copper-induced LDL oxidation depend upon its lipid composition and antioxidant content. AB - Copper promotes oxidation of human low-density lipoprotein (LDL) through molecular mechanisms that are still under investigation. We employed native human LDL, phospholipid-containing delipidated LDL ghosts, or trilinolein reconstituted, phospholipid-containing LDL to investigate both LDL oxidation and the associated process of copper reduction. Both LDL ghosts and trilinolein reconstituted LDL were devoid of antioxidants and were extremely susceptible to AAPH-induced oxidation but, paradoxically, were rather resistant to copper mediated oxidation. The dynamic reduction of Cu(II) to Cu(I) was quantitatively decreased in LDL ghosts and in trilinolein-reconstituted LDL, also lacking the initial rapid reduction and the subsequent inhibition phases, due to the absence of endogenous antioxidants. Conversely, the rate of copper reduction was linear and likely due to lipid peroxides, either already present or formed during copper induced oxidation. We suggest that copper undergoes redox transitions in LDL by utilizing reducing equivalents originating from endogenous antioxidants and/or from lipid peroxides in the LDL lipid core. PMID- 10679290 TI - Frequency and Markov chain analysis of amino-acid sequences of human glutathione reductase. AB - The amino-acid sequence of human glutathione reductase was measured according to two- and three-amino-acid sequences. The measured frequency and probability were compared with predicted frequency and probability. Of 477 two-amino-acid sequences in human glutathione reductase, 176 (36.897%) and 90 (18.868%) sequences can be explained by the predicted frequency and the predicted probability according to a purely random mechanism. Of 477 measured first Markov transition probabilities for the second amino acid in two-amino-acid sequences, 1 (0.210%) measured first Markov transition probability matches the predicted conditional probability and can therefore be explained by a purely random mechanism. No more than two-amino-acid sequences can be explained by a purely random mechanism. PMID- 10679291 TI - Cardiac type cGMP-inhibited phosphodiesterase (PDE3A) gene structure: similarity and difference to adipocyte type PDE3B gene. AB - Phosphodiesterase type 3 isoforms, PDE3A and 3B, are expressed primarily in cardiovascular and adipose tissues, respectively. We previously reported a shorter transcript of 4.4-kb PDE3A which is predominantly transcribed in human placenta, whereas a full-length 7. 6-kb transcript corresponding to the cardiac PDE3A cDNA has not been characterized. Due to unfortunate circumstances created by changes in PDE3 nomenclature, PDE3B gene structure previously reported used PDE3A in its title. Here, we describe PDE3A gene structure, which comprises 16 exons spanning over 130 kb on chromosome 12p12. Two PDE3 isoforms share similar gene organization, but localize to different chromosomes. The most distal transcription initiation site of the PDE3A gene is approximately 1071 bases upstream of the ATG site, suggesting that exon 1 consists of 1071 and 960 bp of untranslated and translated sequences, respectively. The proximal 5'-flanking region, which does not contain TATA-like sequences, exhibited weak but significant promoter activity. Results suggest potential involvement of distal promoter/enhancer and translational regulation for expression of the 7.6-kb transcript. PMID- 10679292 TI - Apolipoprotein E and apolipoprotein D expression in a murine model of singlet oxygen-induced cerebral stroke. AB - Apolipoprotein E (apoE)-deficient mice exhibit neuronal abnormalities similar to those in Alzheimer's disease and enhanced sensitivity to stroke-associated injuries. Here, we show that apoE deficiency results in impaired microglia/macrophage recruitment and accumulation after cerebral infarct. Astrogliosis and apolipoprotein D (apoD) expression are unaffected, suggesting that the neurological abnormalities of apoE-deficient mice could be due to impaired microglia/macrophage recruitment/accumulation, which is important for the clearance of neurodegenerative products via reverse cholesterol transport. To our knowledge, the results presented herein provide the first experimental evidence that brain microglia/macrophage recruitment/accumulation is affected by apoE deficiency. The insights gained from this study should facilitate the elucidation of the role of apoE in neurological disorders such as dementia with stroke and Alzheimer's disease. PMID- 10679293 TI - Survival and differentiation of cultured retinal progenitors transplanted in the subretinal space of the rat. AB - We have shown that embryonic retina contains progenitors which display stem cell properties in vitro. These cells are proliferative and in addition to expressing the neuroectodermal marker, nestin, are multipotential. These properties and the fact that the putative stem cells can differentiate as photoreceptors when exposed to conducive environment identify them as a viable transplantation reagents to address degenerative retinal diseases. Here we report the survival and differentiation of cultured retinal progenitors upon subretinal transplantation. The retinal progenitor grafts, either as neural spheres or in the form of dissociated cells, survived without disrupting the morphology and laminar organization of the host retina. They did not form rosettes, the morphological barrier to the reconstruction of the normal anatomy of the retina. In addition, transplanted progenitors expressed photoreceptor-specific markers, suggesting that progenitors have the potential to differentiate as photoreceptors. Our observations suggest that cultured retinal progenitors can be a viable reagents for therapeutic transplantation. PMID- 10679294 TI - An evaluation of the expression, subcellular localization, and function of rab4 in the exocrine pancreas. AB - The small GTP-binding protein, rab4, is involved in recycling of transferrin receptors and translocation of GLUT4. Recent studies suggest that rab4 controls regulated exocytosis in the exocrine pancreas. We conducted the present study to further investigate the role of rab4 in the exocrine pancreas. We found that the exocrine pancreas expresses two rab4 immunoanalogs, one of approximately 28 kDa identified previously in neonatal glands, and one of approximately 24 kDa which is similar to rab4 characterized in other systems. The latter species was mostly membrane-anchored and localized to endosome-like structures in a supranuclear region that was immunopositive for the transferrin receptor. The approximately 24 kDa rab4 form also localized to the apical plasmamembrane, and this immunofluorescence increased greatly in tissue challenged with a secretagogue. We propose that the approximately 24-kDa rab4 species is involved in compensatory membrane retrieval following regulated exocytosis, and that rab4-positive endocytic vesicles move through a supranuclear recycling compartment. PMID- 10679295 TI - A non-transmembrane form of Jagged-1 regulates the formation of matrix-dependent chord-like structures. AB - Jagged-Notch interactions regulate a transmembrane ligand-receptor signaling pathway involved in the regulation of cell fate determination as well as myoblast and endothelial cell differentiation. To further examine the role of the transmembrane ligand, Jagged-1, in the regulation of cell differentiation, we stably transfected NIH 3T3 cells with a truncated form of Jagged(J)-1, which results in the secretion of a soluble(s) form of the protein. Comparison of gene expression by serial analysis demonstrated that among the 227 transcripts differentially regulated in the sJ-1 transfectants, the expression of the pro alpha-2(I) collagen transcript and pro-alpha-1(I) collagen translation product was predominantly repressed in sJ-1 transfectants. When plated on extracellular matrices, sJ-1 transfectants formed prominent chord-like structures on type I collagen but not on fibrin, fibronectin, or vitronectin. While the sJ-1 transfectants exhibited growth kinetics similar to control cells and were unable to grow in soft agar, the cells were less sensitive to contact inhibition of growth in vitro and sJ-1 allografts formed tissue masses in nude mice after a prolonged latency period and exhibited an abundance of host-derived microvascular endothelial cells. These data suggest that J-1 may be able to modulate, in a matrix-dependent manner, the organization of cell to cell interactions including its ability to promote the development of chord-like structures. PMID- 10679296 TI - Association of EXT1 and EXT2, hereditary multiple exostoses gene products, in Golgi apparatus. AB - We prepared the specific antibodies for EXT1 and EXT2, hereditary multiple exostoses (HME) gene products, and characterized their expression, subcellular localization, and protein association among EXT members. Biochemical analyses indicate that EXT1 and EXT2 can associate and form homo/hetero-oligomers in vivo with or without HME-linked mutations, EXT1 (R340C) and EXT2 (D227N), when exogenously expressed in COS-7 cells. An immunocytochemical analysis showed that both EXT1 and EXT2 localized in Golgi apparatus, irrespective of HME mutations. An immunohistochemical analysis on developing bones further showed that both EXT1 and EXT2 were concomitantly expressed in hypertrophic chondrocytes of forelimb bones from 1-day-old neonatal mouse, but down-regulated in maturing chondrocytes of developing cartilage from 21-day-old mouse. Taken together with the recent finding that EXTs encode for the glycosyltransferase required for the synthesis of heparan sulfate [Lind, T., Tufaro, F., McCormick, C., Lindahl, U., and Lindholt, K. (1998) J. Biol. Chem. 273, 26265-26268], our results implied a molecular basis that a HME-linked mutation found in EXT genes could interfere the physiological function(s) of EXT homo/hetero-oligomers as glycosyltransferases in the developing bones of HME patients. PMID- 10679297 TI - Direct detection of apoptotic cells in peripheral blood from highly pathogenic SHIV-inoculated monkey. AB - Apoptosis in peripheral blood leukocytes (PBL) has been estimated by the enhancement of spontaneous apoptosis after in vitro culture, because apoptotic cells have not been observed directly in freshly isolated PBL in the course of HIV/AIDS. In monkeys infected with a highly pathogenic simian/human immunodeficiency virus (SHIV), which corresponds to rapid progressors of HIV infection, a high frequency of apoptotic cells was directly detected in fresh PBL by electron-microscopic studies. Peripheral blood apoptosis transiently occurred after intense plasma viremia, and peaking at 3 weeks postinfection; occurrence was not limited specifically to lymphocytes, but also occurred in other types of leukocytes. Apoptosis in peripheral lymph nodes was also detected following intense plasma viremia. However, the in vivo apoptosis was not detected in nonpathogenic SHIV-infected monkeys that showed no cell loss. Thus, we directly showed the apoptosis of PBL, which might be associated with pathogenic SHIV produced during the time of plasma viremia. PMID- 10679298 TI - Human growth hormone stimulates proteinase activities of rabbit bone cells via IGF-I. AB - Human growth hormone (hGH) and human insulin-like growth factor-I (hIGF-I) are known to have a marked influence on osteoclastic formation and bone resorption in an unfractionated rabbit bone cell model. This study investigated the effects of both of these factors on the induction of cysteine-proteinases and matrix metalloproteinase-2 (MMP-2) and MMP-9. After 4 days of rabbit bone cell culture, hGH and hIGF-I significantly modulated cathepsin, MMP-9 (latent form) and MMP-2 (active form) activities. Similar studies were performed in the presence of parathyroid hormone (hPTH). hPTH increased MMP-2 and MMP-9 activities whereas it had no effect on the production of cathepsins by bone cells. When neutralizing anti-hIGF-1 antiserum was added to the culture, the stimulatory effects of hGH were totally abolished, indicating that hGH-modulated cathepsin and metalloproteinase activities were partly mediated by local hIGF-I secretion. Cysteine-proteinase activities released by purified osteoclasts were very low and were not modulated by hGH and h-IGF-I. However, hIGF-I but not hGH increased MMP 2 and MMP-9 activities released by purified osteoclasts. It may be concluded that hGH markedly stimulates the expression of proteinases in total rabbit bone cells via local hIGF-I production by stromal cells. Cysteine-proteinase activities are mainly produced by non-osteoclastic cells, while MMP-2 and MMP-9 modulated by hIGF-I are mainly expressed by osteoclastic cells. PMID- 10679299 TI - Binding domain for p21(WAF1) on the polypeptide chain of the protein kinase CK2 beta-subunit. AB - Protein kinase CK2 is a ubiquitous serine/threonine kinase which is involved in many proliferation-related processes in the cell. It is composed of two regulatory beta-subunits and two catalytic alpha-subunits. Its regulation still remains mysterious in spite of many years of intense research. One of its regulators is the cdk inhibitory molecule p21(WAF1)-a protein which is expressed in situations of genotoxic stress. p21(WAF1) binds to the beta-subunit of CK2 and inhibits the activity of CK2. Using deletion mutants of CK2 beta as well as a peptide library consisting of 15-amino-acid-long peptides derived from the polypeptide chain of CK2 beta we mapped the binding region for p21(WAF1) on the polypeptide chain of CK2 beta. We localized an amino-terminal and a carboxy terminal binding domain. Binding of p21(WAF1) to both regions of the CK2 beta subunit interferes with the phosphotransferase activity of the CK2 holoenzyme. PMID- 10679300 TI - Promoter usage for insulin-like growth factor-II in cancerous and benign human breast, prostate, and bladder tissues, and confirmation of a 10th exon. AB - Upregulation of insulin-like growth factor (IGF)-II expression has been reported for a variety of childhood and adulthood tumors. We determined IGF-II gene promoter usage in human cancerous and benign tissues by semiquantitative RT-PCR using P1-P4-specific primers. Although the human IGF-II gene structure is commonly thought to consist of nine exons and four promoters, we detected substantial utilization of a previously reported exon 4b, which is downstream of exon 4. Thus, exon 4b was intensively studied using 4b-specific primers. IGF-II gene promoter usage is highly variable in malignant and benign breast, prostate, and bladder tissues. While a majority of samples utilized P2-P4 promoters in a variety of combinations, when quantitated, P3 and P4 promoters were much more active than P2 promoter. This study not only demonstrated that IGF-II gene promoter usage is highly variable in malignant and benign tissues, but suggested that alternatively spliced exon 4b should be recognized as a 10th exon. PMID- 10679301 TI - Three rat brain alternative splicing dynamin-like protein variants: interaction with the glycogen synthase kinase 3beta and action as a substrate. AB - Dynamin-like protein, a large GTP-binding protein, has recently been cloned, and studies have shown that it may be involved in the formation of coated vesicles. In this report, three different alternatively spliced dynamin-like protein variants (DLP1-WT, -11, and -37) from rat brain were identified by reverse transcription/polymerase chain reaction (RT-PCR). One novel rat alternatively spliced variant (DLP1-37), not described previously, was identified. We examined the interaction of these three rat brain dynamin-like protein variants with glycogen synthase kinase 3beta (Gsk-3beta) using the yeast two-hybrid screening, in vitro binding assay, and immunoprecipitation analysis. It was found that all three examined rat brain dynamin-like protein variants can bind to Gsk-3beta. Moreover, in vitro kinase (phosphorylation) assay showed that mammalian dynamin like protein acts as a substrate for glycogen synthase kinase 3beta. These data suggest that Gsk-3beta may participate in a functional role in dynamin-like proteins in vesicle trafficking. PMID- 10679302 TI - High extracellular calcium concentrations directly stimulate osteoclast apoptosis. AB - Although the inhibitory effects of high extracellular calcium concentrations ([Ca](e)) on osteoclastic bone resorption have been known for several years, the exact mechanism remains poorly understood. The present study was performed to investigate the possible effect of [Ca](e) on osteoclast apoptosis. Using highly purified rabbit osteoclasts, we have shown that calcium directly promotes apoptosis in a dose-dependent manner which correlates with the dose range of calcium for the inhibition of bone resorption. A time-course experiment of apoptotic changes of osteoclasts cultured in presence of 1.8 or 20 mM calcium showed a significant difference after as early as 8 h of culture. After 72 h of culture, we observed that 80% of the cells cultured in the presence of 20 mM calcium displayed the typical features of apoptosis compared to only 20% in the medium containing 1.8 mM calcium. Calcium channel blockers and ryanodine abrogated the effects of [Ca](e) on apoptosis while neomycin, a calcium-sensing receptor agonist, did not alter cell viability. Taken together, these results suggest that calcium influx is involved in calcium-induced osteoclast apoptosis. Our results are consistent with the concept that in the presence of high [Ca](e) generated during bone demineralization, osteoclasts are subjected to negative feedback regulation due, at least in part, to the induction of apoptosis. PMID- 10679303 TI - Characterization of endopeptidase activity of tripeptidyl peptidase-I/CLN2 protein which is deficient in classical late infantile neuronal ceroid lipofuscinosis. AB - Endopeptidase activities of the CLN2 gene product (Cln2p)/tripeptidyl peptidase I (TPP-I), purified from rat spleen, were studied using the synthetic fluorogenic substrates. We designed and constructed decapeptides, based on the known sequence cleavage specificities of bacterial pepstatin-insensitive carboxyl proteases (BPICP). MOCAc-Gly-Lys-Pro-Ile-Pro-Phe-Phe-Arg-Leu-Lys(Dnp)r-NH(2) is readily hydrolyzed by Cln2p/TPP-I (K(cat)/K(m) = 7.8 s(-1) mM(-1)). The enzyme had a maximal activity at pH 3.0 for an endopeptidase substrate, but at pH 4.5 with respect to tripeptidyl peptidase activity. Both endopeptidase and tripeptidyl peptidase activities were strongly inhibited by Ala-Ala-Phe-CH(2)Cl, but not inhibited by tyrostatin, an inhibitor of bacterial pepstatin-insensitive carboxyl proteases, pepstatin, or inhibitors of serine proteases. Fibroblasts from classical late infantile neuronal ceroid lipofuscinosis patients have less than 5% of the normal tripeptidyl peptidase activity and pepstatin-insensitive endopeptidase activity. Cln2p/TPP-I is a unique enzyme with both tripeptidyl peptidase and endopeptidase activities for certain substrate specificity. PMID- 10679304 TI - In vitro binding of nucleolin to double-stranded telomeric DNA. AB - We have purified a 100 kDa protein, resolved in a Southwestern binding screen of total nuclear proteins from Hela cells with double-stranded human telomeric probe. A polyclonal antiserum raised by this protein recognizes purified nucleolin and stains nucleoli in growing Hela cells. We demonstrate that a truncated form of human nucleolin and a purified deletion derivative of mouse nucleolin bind in vitro to duplex telomeric DNA. This study suggests a new link between telomeres and the nucleolus. PMID- 10679305 TI - MPP(+)-induced mitochondrial dysfunction is potentiated by dopamine. AB - MPP(+), the major metabolite of the Parkinsonism-inducing compound MPTP, responsible for the destruction of the nigrostriatal pathway in primates and rodents, has been assayed in isolated rat liver mitochondria in the presence of physiological concentrations of dopamine or analogous concentrations of melanin dopamine. 5 microM MPP(+) in the presence of 70 microM dopamine or melanin dopamine, but not alone, decreased the heat production and oxygen consumption of a mitochondrial suspension activated with succinate and ADP. Both dopamine and oxidized dopamine plus MPP(+) also decreased the mitochondrial reductive power measured with MTT. Mitochondrial swelling was observed, associated with an increase in membrane mitochondrial potential, as a synergistic effect between low concentrations of MPP(+) and dopamine. It is suggested that cytosolic dopamine, by itself or via its autooxidation products, may play a relevant role in the mitochondrial toxicity of MPP(+). A failure in the regulation of the storage/release of dopamine could aggravate a mitochondrial damage and trigger the neurodegenerative process underlying MPTP toxicity and Parkinson's disease. PMID- 10679306 TI - Mouse peroxiredoxin V is a thioredoxin peroxidase that inhibits p53-induced apoptosis. AB - We have identified human and mouse peroxiredoxin V (Prx-V) by virtue of the sequence homologies to yeast peroxisomal antioxidant enzyme PMP20. Prx-V represents the fifth of the six currently known subfamilies of mammalian peroxiredoxins. It is a novel organellar enzyme that has orthologs in bacteria. Biochemically, Prx-V is a thioredoxin peroxidase. One important aspect of p53 function in mammalian cells involves induction of apoptosis likely mediated by redox. We show that overexpression of Prx-V prevented the p53-dependent generation of reactive oxygen species. Likewise, Prx-V inhibited p53-induced apoptosis. Thus, Prx-V is critically involved in intracellular redox signaling. PMID- 10679307 TI - Induction of COX-2 expression by nitric oxide in rheumatoid synovial cells. AB - Prostaglandins formed by cyclooxygenase (COX) enzymes are important mediators of inflammation. The contribution of inducible COX-2 in the rheumatoid synovium is well documented. In this study, we evaluated the contribution of nitric oxide (NO) to COX-2 expression in rheumatoid synovial cells. Exposure of rheumatoid synovial cells to a NO donor, SNAP, induced COX-2 protein expression in a dose dependent manner. RT-PCR analysis also demonstrated that COX-2 mRNA was induced in SNAP-treated synovial cells. Dexamethasone at therapeutic concentrations markedly inhibited this NO-mediated COX-2 expression in synovial cells. In contrast to its effect on COX-2 expression, SNAP did not affect the constitutive expression of COX-1 in rheumatoid synovial cells. Our findings suggest that NO is an important modulator of COX-2 expression and that glucocorticoids exert their anti-inflammatory action in rheumatoid synovium, at least in part, by suppression of COX-2 induction. PMID- 10679308 TI - Cloning and characterization of a gene, mpsA, encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1. AB - Proteins located within the lipid bilayer, surrounding the intracellular bacterial magnetic particles (BMP) from Magnetospirillum sp. AMB-1, were separated using SDS-PAGE. Several major proteins of approximate molecular weight 66.2, 35.6, and 24.8 kDa were identified. The N-terminal amino acid sequence of one of these proteins, designated MpsA, was determined and used to design a pair of PCR primers which amplified a 105 bp DNA fragment from AMB-1 genomic DNA. Gene walking, using anchored PCR, was used to determine the complete nucleotide sequence (954 bp) of the mpsA gene. The mpsA encodes a 317 amino acid protein which does not have an N-terminal cytoplasmic transport signal sequence. Intracellular localization studies were carried out using an mpsA-luc gene fusion expressed in AMB-1 following gene transfer by conjugation. The gene fusion was constructed by cloning a 1.6 kb mpsA fragment upstream of luc in the conjugal plasmid pKLC. The MpsA-Luc fusion protein was preferentially located on the magnetic particle membrane. Although the function of MpsA remains unknown, homology searches suggest similarity with the alpha subunit of acetyl-CoA carboxylase and the CoA-binding motif. PMID- 10679309 TI - Identification of viral macrophage inflammatory protein (vMIP)-II as a ligand for GPR5/XCR1. AB - Lymphotactin is unique among chemokines in that it contains only two of four conserved cysteines and may possess a structure less constrained than other chemokines. The viral chemokine vMIP-II, which presumably has a structure similar to that of CC chemokines has been shown to inhibit many chemokine receptors, but its activity at GPR5/XCR1 has not been described. Interestingly, vMIP-II (but not vMIP-I) was found to be a potent antagonist of lymphotactin activity at GPR5/XCR1, extending the range of chemokine classes that this viral protein is known to inhibit to include the C class chemokine. In addition, we have extended previous analyses of GPR5/XCR1 expression and show that this receptor is expressed in leukocyte cells previously shown to be responsive to lymphotactin. PMID- 10679310 TI - Pyrrolidine dithiocarbamate induces apoptosis by a cytochrome c-dependent mechanism. AB - Pyrrolidine dithiocarbamate (PDTC) is a synthetic antioxidant molecule, which has been recently proposed as an antitumoral agent on the basis of its capability of inducing apoptosis. We investigated the effect of PDTC on the proliferation and survival of the promyelocitic cell line HL-60. Concentration as low as 10 microM of PDTC induces a significant reduction of the growth rate and the contemporaneous activation of the apoptotic process. Programmed cell death was demonstrated by biochemical analyses, including the activation of procaspase 3 and the cleavage of poly(ADP-ribose) polymerase (PARP). PDTC-dependent apoptosis was associated with an early release of cytochrome c from mitochondria, while the involvement of pathways due to cell death receptors engagement was ruled out by detailed time-course analyses of caspases 3 and 8 activation. Moreover, no up regulation of p21(CIP1) level, a pivotal cyclin-dependent kinase inhibitor, occurred at PDTC concentration able to induce apoptosis. Finally, in vitro incubation of purified mitochondria with PDTC demonstrated that the molecule is directly able to induce cytochrome c release from the intermembrane space, thus confirming that mitochondria are a primary cellular target of the molecule. PMID- 10679312 TI - Epstein-Barr virus: Co-opting B-cell memory and migration. AB - Epstein-Barr virus, a B-lymphotropic human herpesvirus, persists in vivo by entering the long-lived memory B-cell compartment. Work with genetically modified mice suggests that the viral latent membrane protein LMP1 might allow infected B cells to access the memory compartment by an unusual route. PMID- 10679311 TI - Xenopus and chicken sperm contain a cytosolic soluble protein factor which can trigger calcium oscillations in mouse eggs. AB - There is evidence showing that at fertilization the sperm introduces into egg cytoplasm a protein-based cytosolic factor, which serves as the physiological trigger for inducing Ca(2+) oscillations in mammalian eggs. Here we show that sperm of nonmammalian vertebrates also contain a cytosolic protein factor that can induce Ca(2+) oscillations when introduced into mammalian eggs. We have observed that cytosolic extracts derived from Xenopus or chicken sperm could induce mouse eggs to undergo Ca(2+) oscillations similar to those induced by bovine sperm extracts. The factor responsible for inducing Ca(2+) oscillations was of high molecular weight and heat- or proteinase K-labile. We show that 0.5 chicken sperm-equivalents or 1-2 Xenopus sperm-equivalents of the extracts had enough activity to trigger Ca(2+) oscillations in mouse eggs. Our findings illustrate that although Xenopus, chicken, and mammals are evolutionarily divergent species, the function of the sperm protein factor in triggering Ca(2+) oscillations in mammalian eggs appears not to be species specific in vertebrates. PMID- 10679313 TI - Axon regeneration: Vaccinating against spinal cord injury. AB - Myelin is a potent inhibitor of axon regeneration, but has been viewed as just one of many factors that prevent regeneration after injury. So it comes as a surprise that immunization against myelin has been found to allow extensive axon regeneration after injury, without apparent autoimmune-induced demyelination. PMID- 10679314 TI - Neurotransmission: Chemical and electrical interneuron coupling. AB - Recent studies have described the coupling between pairs of neocortical interneurons involving both electrical and chemical transmission; these new results may have important implications for the mechanisms underlying neuronal synchrony and rhythmic activity in the brain. PMID- 10679315 TI - Seed plant phylogeny: Demise of the anthophyte hypothesis? AB - Recent molecular phylogenetic studies indicate, surprisingly, that Gnetales are related to conifers, or even derived from them, and that no other extant seed plants are closely related to angiosperms. Are these results believable? Is this a clash between molecules and morphology? PMID- 10679316 TI - Protein folding: Thickening the broth. AB - Recent results support the notion that macromolecular 'crowding' enhances protein aggregation, at the expense of correct folding. The results can be rationalised in terms of kinetic competition between distinct processes, taking into account the relative influence of crowding on each process. PMID- 10679317 TI - Intracellular signalling: Is PIP(2) a messenger too? AB - The phospholipid phosphatidylinositol (4,5) bisphosphate (PIP(2)) has recently been shown to act downstream of the small G proteins Rac and Arf. Different effectors may be employed in each case, suggesting that PIP(2) has multiple signalling roles. PMID- 10679318 TI - Signal transduction: Life, the universe and ... development. AB - Glycogen synthase kinase 3 (GSK-3) is a key component of the Wnt signalling pathway, among others, and is known to be regulated by inhibition. Now a novel, dual specificity protein kinase known as Zaphod kinase has been discovered that activates GSK-3 by tyrosine phosphorylation. PMID- 10679319 TI - Quantitative genetics: Resolving wing shape genes. AB - Recent high-resolution quantitative mapping experiments aimed at elucidating the genetics of natural variation for wing shape in fruit-flies suggest that very many genes can subtly influence the trait. PMID- 10679320 TI - Molecular motors: Kinesin's dynamically dockable neck. AB - Kinesin is a molecular walking machine with two identical motor heads connected to a coiled-coil tail. Details of the coordination mechanism, which causes kinesin to walk directionally, and the tracking mechanism, which guides each detaching head to its next site on the microtubule, are beginning to emerge. PMID- 10679321 TI - Activating the DNA damage checkpoint in a developmental context. AB - BACKGROUND: Studies in unicellular systems have established that DNA damage by irradiation invokes a checkpoint that acts to stall cell division. During metazoan development, the modulation of cell division by checkpoints must occur in the context of gastrulation, differential gene expression and changes in cell cycle regulation. To understand the effects of checkpoint activation in a developmental context, we examined the effect of X-rays on post-blastoderm embryos of Drosophila melanogaster. RESULTS: In Drosophila, DNA damage was previously found to delay anaphase chromosome separation during cleavage cycles that lack a G2 phase. In post-blastoderm cycles that included a G2 phase, we found that irradiation delayed the entry into mitosis. Gastrulation and the developmental program of string (Cdc25) gene expression, which normally regulates the timing of mitosis, occurred normally after irradiation. The radiation-induced delay of mitosis accompanied the exclusion of mitotic cyclins from the nucleus. Furthermore, a mutant form of the mitotic kinase Cdk1 that cannot be inhibited by phosphorylation drove a mitotic cyclin into the nucleus and overcame the delay of mitosis induced by irradiation. CONCLUSIONS: Developmental changes in the cell cycle, for example, the introduction of a G2 phase, dictate the response to checkpoint activation, for example, delaying mitosis instead of or in addition to delaying anaphase. This unprecedented finding suggests that different mechanisms are used at different points during metazoan development to stall cell division in response to checkpoint activation. The delay of mitosis in post-blastoderm embryos is due primarily to inhibitory phosphorylation of Cdk1, whereas nuclear exclusion of a cyclin-Cdk1 complex might play a secondary role. Delaying cell division has little effect on gastrulation and developmentally regulated string gene expression, supporting the view that development generally dictates cell proliferation and not vice versa. PMID- 10679322 TI - Rsk1 mediates a MEK-MAP kinase cell survival signal. AB - BACKGROUND: Growth factors activate an array of cell survival signaling pathways. Mitogen-activated protein (MAP) kinases transduce signals emanating from their upstream activators MAP kinase kinases (MEKs). The MEK-MAP kinase signaling cassette is a key regulatory pathway promoting cell survival. The downstream effectors of the mammalian MEK-MAP kinase cell survival signal have not been previously described. RESULTS: We identify here a pro-survival role for the serine/threonine kinase Rsk1, a downstream target of the MEK-MAP kinase signaling pathway. In cells that are dependent on interleukin-3 (IL-3) for survival, pharmacological inhibition of MEKs antagonized the IL-3 survival signal. In the absence of IL-3, a kinase-dead Rsk1 mutant eliminated the survival effect afforded by activated MEK. Conversely, a novel constitutively active Rsk1 allele restored the MEK-MAP kinase survival signal. Experiments in vitro and in vivo demonstrated that Rsk1 directly phosphorylated the pro-apoptotic protein Bad at the serine residues that, when phosphorylated, abrogate Bad's pro-apoptotic function. Constitutively active Rsk1 caused constitutive Bad phosphorylation and protection from Bad-modulated cell death. Kinase-inactive Rsk1 mutants antagonize Bad phosphorylation. Bad mutations that prevented phosphorylation by Rsk1 also inhibited Rsk1-mediated cell survival. CONCLUSIONS: These data support a model in which Rsk1 transduces the mammalian MEK-MAP kinase signal in part by phosphorylating Bad. PMID- 10679323 TI - Ordered assembly of roX RNAs into MSL complexes on the dosage-compensated X chromosome in Drosophila. AB - BACKGROUND: In the male Drosophila, the X chromosome is transcriptionally upregulated to achieve dosage compensation, in a process that depends on association of the MSL proteins with the X chromosome. A role for non-coding RNAs has been suggested in recent studies. The roX1 and roX2 RNAs are male-specific, non-coding RNAs that are produced by, and also found associated with, the dosage compensated male X chromosome. Whether roX RNAs are physically part of the MSL complex has not been resolved. RESULTS: We found that roX RNAs colocalize with the MSL proteins and are highly unstable unless the MSL complex is coexpressed, suggesting a physical interaction. We were able to immunoprecipitate roX2 RNA from male tissue-culture cells with antibodies to the proteins Msl1 and Mle, consistent with an integral association with MSL complexes. Localization of roX1 and roX2 RNAs in mutants indicated an order of MSL-complex assembly in which roX2 RNA is incorporated early in a process requiring the Mle helicase. We also found that the roX2 gene, like roX1, is a nucleation site for MSL complex spreading into flanking chromatin in cis. CONCLUSIONS: Our results support a model in which MSL proteins assemble at specific chromatin entry sites (including the roX1 and roX2 genes); the roX RNAs join the complex at their sites of synthesis; and complete complexes spread in cis to dosage compensate most genes on the X chromosome. PMID- 10679324 TI - Type Ialpha phosphatidylinositol-4-phosphate 5-kinase mediates Rac-dependent actin assembly. AB - Action polymerization is essential for a variety of cellular processes including movement, cell division and shape change. The induction of actin polymerization requires the generation of free actin filament barbed ends, which results from the severing or uncapping of pre-existing actin filaments [1] [2], or de novo nucleation, initiated by the Arp2/3 complex [3] [4] [5] [6] [7]. Although little is known about the signaling pathways that regulate actin assembly, small GTPases of the Rho family appear to be necessary [8] [9] [10] [11]. In thrombin stimulated platelets, the Rho family GTPase Rac1 induces actin polymerization by stimulating the uncapping of actin filament barbed ends [2]. The mechanism by which Rac regulates uncapping is unclear, however. We previously demonstrated that Rac interacts with a type I phosphatidylinositol-4-phosphate 5-kinase (PIP 5 kinase) in a GTP-independent manner [12] [13]. Because PIP 5-kinases synthesize phosphatidylinositol-4,5-bisphosphate (PI(4,5)P(2)), a lipid that dissociates capping proteins from the barbed ends of actin filaments [14] [15] [16], they are good candidates for mediating the effects of Rac on actin assembly. Here, we have identified the Rac-associated PIP 5-kinase as the PIP 5-kinase isoforms alpha and beta. When added to permeabilized platelets, PIP 5-kinase alpha induced actin filament uncapping and assembly. In contrast, a kinase-inactive PIP 5-kinase alpha mutant failed to induce actin assembly and blocked assembly stimulated by thrombin or Rac. Furthermore, thrombin- or Rac-induced actin polymerization was inhibited by a point mutation in the carboxyl terminus of Rac that disrupts PIP 5 kinase binding. These results demonstrate that PIP 5-kinase alpha is a critical mediator of thrombin- and Rac-dependent actin assembly. PMID- 10679325 TI - Role of proton gradients and vacuola H(+)-ATPases in the refilling of intracellular calcium stores in exocrine cells. AB - Numerous hormones and neurotransmitters activate cells by increasing cytosolic calcium concentration ([Ca(2+)](i)), a key regulatory factor for many cellular processes. A pivotal feature of these Ca(2+) signals is the release of Ca(2+) from intracellular stores, which is followed by activation of extracellular calcium influx, allowing refilling of the stores by SERCA pumps associated with the endoplasmic reticulum. Although the mechanisms of calcium release and calcium influx have been extensively studied, the biology of the Ca(2+) stores is poorly understood. The presence of heterogeneous calcium pools in cells has been previously reported [1] [2] [3]. Although recent technical improvements have confirmed this heterogeneity [4], knowledge about the mechanisms underlying Ca(2+) transport within the stores is very scarce and rather speculative. A recent study in polarized exocrine cells [5] has revealed the existence of Ca(2+) tunneling from basolateral stores to luminal pools, where Ca(2+) is initially released upon cell activation. Here, we present evidence that, during stimulation, Ca(2+) transported into basolateral stores by SERCA pumps is conveyed toward the luminal pools driven by proton gradients generated by vacuolar H(+)-ATPases. This finding unveils a new aspect of the machinery of Ca(2+) stores. PMID- 10679326 TI - Role of the kinesin neck linker and catalytic core in microtubule-based motility. AB - Kinesin motor proteins execute a variety of intracellular microtubule-based transport functions [1]. Kinesin motor domains contain a catalytic core, which is conserved throughout the kinesin superfamily, followed by a neck region, which is conserved within subfamilies and has been implicated in controlling the direction of motion along a microtubule [2] [3]. Here, we have used mutational analysis to determine the functions of the catalytic core and the approximately 15 amino acid 'neck linker' (a sequence contained within the neck region) of human conventional kinesin. Replacement of the neck linker with a designed random coil resulted in a 200-500-fold decrease in microtubule velocity, although basal and microtubule stimulated ATPase rates were within threefold of wild-type levels. The catalytic core of kinesin, without any additional kinesin sequence, displayed microtubule stimulated ATPase activity, nucleotide-dependent microtubule binding, and very slow plus-end-directed motor activity. On the basis of these results, we propose that the catalytic core is sufficient for allosteric regulation of microtubule binding and ATPase activity and that the kinesin neck linker functions as a mechanical amplifier for motion. Given that the neck linker undergoes a nucleotide-dependent conformational change [4], this region might act in an analogous fashion to the myosin converter, which amplifies small conformational changes in the myosin catalytic core [5,6]. PMID- 10679327 TI - Lif1p targets the DNA ligase Lig4p to sites of DNA double-strand breaks. AB - DNA ligases catalyse the joining of DNA single- and double-strand breaks. Saccharomyces cerevisiae Cdc9p is a homologue of mammalian DNA ligase I and is required for DNA replication, recombination and single-strand break repair. The other yeast ligase, Lig4p/Dnl4p, is a homologue of mammalian DNA ligase IV, and functions in the non-homologous end-joining (NHEJ) pathway of DNA double-strand break repair [1] [2] [3] [4]. Lig4p interacts with Lif1p, the yeast homologue of the human ligase IV-associated protein, XRCC4 [5]. This interaction takes place through the carboxy-terminal domain of Lig4p and is required for Lig4p stability. We show that the carboxy-terminal interaction region of Lig4p is necessary for NHEJ but, when fused to Cdc9p, is insufficient to confer NHEJ function to Cdc9p. Also, Lif1p stimulates the in vitro catalytic activity of Lig4p in adenylation and DNA ligation. Nevertheless, Lig4p is inactive in NHEJ in the absence of Lif1p in vivo, even when Lig4p is stably expressed. We show that Lif1p binds DNA in vitro and, through in vivo cross-linking and chromatin immuno precipitation assays, demonstrate that it targets Lig4p to chromosomal DNA double-strand breaks. Furthermore, this targeting requires another key NHEJ protein, Ku. PMID- 10679328 TI - Discrete in vivo roles for the MutL homologs Mlh2p and Mlh3p in the removal of frameshift intermediates in budding yeast. AB - The DNA mismatch repair machinery is involved in the correction of a wide variety of mutational intermediates. In bacterial cells, homodimers of the MutS protein bind mismatches and MutL homodimers couple mismatch recognition to downstream processing steps [1]. Eukaryotes possess multiple MutS and MutL homologs that form discrete, heterodimeric complexes with specific mismatch recognition and repair properties. In yeast, there are six MutS (Msh1-6p) and four MutL (Mlh1-3p and Pms1p) family members [2] [3]. Heterodimers comprising Msh2p and Msh3p or Msh2p and Msh6p recognize mismatches in nuclear DNA [4] [5] and the subsequent processing steps most often involve a Mlh1p-Pms1P heterodimer [6] [7]. Mlh1p also forms heterodimeric complexes with Mlh2p and Mlh3p [8], and a minor role for Mlh3p in nuclear mismatch repair has been reported [9]. No mismatch repair function has yet been assigned to the fourth yeast MutL homolog, Mlh2p, although mlh2 mutants exhibit weak resistance to some DNA damaging agents [10]. We have used two frameshift reversion assays to examine the roles of the yeast Mlh2 and Mlh3 proteins in vivo. This analysis demonstrates, for the first time, that yeast Mlh2p plays a role in the repair of mutational intermediates, and extends earlier results implicating Mlh3p in mismatch repair. PMID- 10679329 TI - Re-modelling of nuclear architecture in quiescent and senescent human fibroblasts. AB - Spatial organisation of the genome within the nucleus can play a role in maintaining the expressed or silent state of some genes [1]. There are distinct addresses for specific chromosomes, which have different functional characteristics, within the nuclei of dividing populations of human cells [2]. Here, we demonstrate that this level of nuclear architecture is altered in cells that have become either quiescent or senescent. Upon cell cycle exit, a gene-poor human chromosome moves from a location at the nuclear periphery to a more internal site in the nucleus, and changes its associations with nuclear substructures. The chromosome moves back toward the edge of the nucleus at a distinctive time after re-entry into the cell cycle. There is a 2-4 hour period at the beginning of G1 when the spatial organisation of these human chromosomes is established. Lastly, these experiments provide evidence that temporal control of DNA replication can be independent of spatial chromosome organisation. We conclude that the sub-nuclear organisation of chromosomes in quiescent or senescent mammalian somatic cells is fundamentally different from that in proliferating cells and that the spatial organisation of the genome is plastic. PMID- 10679330 TI - Biology in pictures. Grow with the flow. PMID- 10679331 TI - The yen for success. PMID- 10679332 TI - Allergy. PMID- 10679333 TI - Fevered headlines. PMID- 10679334 TI - Muscular dystrophy. PMID- 10679335 TI - Chemoinformatics - similarity and diversity in chemical libraries. AB - Molecular similarity and molecular diversity techniques lie at the heart of attempts to design structurally diverse combinatorial libraries for the identification of novel bioactive compounds. Recent advances include the development of new types of selection algorithm, the validation of such algorithms, the use of filtering systems to screen out undesirable molecules prior to the design of a library, and the integration of similarity and diversity analysis with other methods for computer-aided molecular design. PMID- 10679336 TI - Neutron scattering: good news for biotechnology. AB - In its application to biological systems, neutron scattering is still an emerging technology with a great deal of potential. A consequence of the native interaction between neutrons and biological samples is that the hydrogen isotopes (1)H and (2)H are most significant in dynamical and structural studies, respectively. PMID- 10679337 TI - Large-scale predictions of secretory proteins from mammalian genomic and EST sequences. AB - Machine learning techniques have improved predictions of secretory proteins from protein, genomic and expressed sequence tag (EST) sequences. Artificial neural networks, physical sequence analysis using high-performance optimization, and hidden Markov models identify extremely variable signal peptides (the vehicles of protein transport across the endoplasmic reticulum membrane), transmembrane segments, and specific extracellular and intracellular domains as indicators of possible roles in the intercellular and intracellular chemical signaling pathways. The major role of peptide hormones, blood coagulation factors, carcinogenesis agents, and other secretory proteins in orchestrating multicellular life indicates pharmacological potential in the cure of major diseases and numerous biotechnological applications. PMID- 10679338 TI - Extraction of pharmacophore information from high-throughput screens. AB - Two major advances have been made in the computational perception and utilization of pharmacophores in compound libraries, both real and virtual. Firstly, a hierarchical set of filtering calculations has emerged that can be used to efficiently partition a library into a trial set of pharmacophores. This sequential filtering permits large libraries to be efficiently processed, as well as compounds judged as 'hits' to be analyzed in great detail. Secondly, new and extended methods of QSAR (quantitative structure-activity relationship) analysis have evolved to translate pharmacophore information into QSAR models that, in turn, can be used as virtual high-throughput screens for activity profiling of a library. PMID- 10679339 TI - Microarray technology - enhanced versatility, persistent challenge. AB - Microarray analysis of nucleic acid related phenomena on a genome-wide scale is now a proven technology. New applications of the method are appearing rapidly and problems unique to the handling and interpretation of the large data sets produced by the technique are beginning to be addressed. PMID- 10679340 TI - Visualization of data. AB - Data visualization has developed in several directions: theoretical; methodological; and in new application areas. Advances include the development of a grammar of graphics, deeper understanding of human perception and implications for graphical layout, and better approaches to visualizing multi-dimensional data and large data sets. Gene expression is a notable new application area for visualization of large data sets. PMID- 10679341 TI - Finding function through structural genomics. AB - The recent availability of whole-genome sequences and large numbers of protein coding regions from high-throughput cDNA analysis has fundamentally changed experimental biology. These efforts have provided huge databases of protein sequences, many of which are of unknown function. Deciphering the functions of these myriad proteins presents a major intellectual challenge. PMID- 10679342 TI - Advances in surface plasmon resonance biosensor analysis. AB - The number and diversity of surface plasmon resonance (SPR) biosensor applications continue to increase. Evolutions in instrument and sensor chip technology, experimental methodology, and data analysis are making it possible to examine a wider variety of biomolecular interactions in greater mechanistic detail. SPR biosensors are poised to make a significant impact in basic research and pharmaceutical discovery. PMID- 10679343 TI - Discovery and modeling of transcriptional regulatory regions. AB - A complex network of regulatory controls governs the patterns of gene expression. Enabled by the tools of molecular cloning, initial experimental queries into the gene regulatory network elucidated a wide array of transcription factors and their cognate binding sites from hundreds of genes. The recent fusion of genome scale experimental tools, a more comprehensive gene catalog, and concomitant advances in computational methodology, has extended the range of questions being posed. The potential to further our understanding of the biochemical mechanisms of transcriptional regulation and to accelerate the delineation of regulatory control regions in the human genome is enormous. PMID- 10679344 TI - Chemoinformatics - predicting the physicochemical properties of 'drug-like' molecules. AB - A few major advances have occurred in the area of physicochemical modeling of organic compounds during the past several years, spurred on by changes in the pharmaceutical industry. Recent advances include the ability to categorize and screen the overall physicochemical properties of potential drug candidates based entirely on their molecular structures and the ability to model the components that contribute to the oral absorption characteristics of potential drug candidates. PMID- 10679345 TI - Structural energetics of protein folding and binding. AB - Structural energetics is a method for calculating the energetics of protein folding and binding reactions as a function of temperature. This approach allows measured energetics to be interpreted with regards to the protein structure and the prediction of energetics from known structures. Recent advances include improvements in the parameterization of enthalpy, entropy and heat capacity terms and new applications, especially with regards to understanding dynamic properties of proteins and how these are affected by ligand binding. PMID- 10679346 TI - Biotechnology match making: screening orphan ligands and receptors. AB - To date there has been a considerable amount of interest and success in the pharmaceutical industry in the discovery of drug targets and diagnostics via genomic technologies, namely DNA sequencing, mutation/polymorphism detection and expression monitoring of mRNA. As the ultimate targets for the majority of these methods are actually proteins, more and more emphasis has been placed upon protein-based methods in an effort to define the function of proteins discovered by genomic technologies. One of the most challenging areas of drug target discovery facing researchers today is the search for novel receptor-ligand pairs. Database mining techniques in conjunction with other computational methods are able to identify many novel sequences of putative receptors, but the ability to similarly identify the receptor's natural ligand is not possible by these methods. The past few years have seen an increase in methodology and instrumentation focused on the ability to discover and characterize protein protein interactions, as well as receptor-ligand pairs. Significant advances have been made in the areas of instrumentation (biosensors and fluorescent plate readers) as well as methodologies relating to phage/ribosome display and library construction. PMID- 10679347 TI - Riding the wave: structural and energetic principles of helical membrane proteins. AB - Genome sequencing efforts have revealed that perhaps as many as 20-40% of open reading frames in complex organisms may encode proteins containing at least one helical transmembrane segment. Contrasting with this approaching tidal wave of helical membrane proteins is the fact that our understanding of the sequence structure-function relationships for membrane proteins lags far behind that of soluble proteins. This looming reality emphasizes the tremendous biochemical and structural work that remains to be done on helical membrane proteins in order to elucidate the structural and energetic principles that specify and stabilize their folds, which define their functions. These facts are not lost on the pharmaceutical industry, where successful therapeutics and major discovery efforts are targeting membrane proteins. PMID- 10679348 TI - Biopharmaceutical formulation. AB - The rapid maturation of the field of biopharmaceutical formulation is the result of the simultaneous development of a thermodynamic mechanism for protein-solvent interaction and identification of natural chemicals employed by nature to stabilize proteins in response to environmental stresses. In general, these cosolvents are excluded from the protein surface. Proteins are maintained in their native folded conformations by these cosolvents as a result of the highly unfavorable interaction between cosolvents and peptide backbones, which would be exposed to the cosolvent upon unfolding. PMID- 10679349 TI - High-throughput and ultra-high-throughput screening: solution- and cell-based approaches. AB - The trend towards assay miniaturization for high-throughput and ultra-high throughput screening continues to spur development of homogeneous, fluorescence based assays in higher density, smaller volume microplate formats. Recently, first-generation microfluidic devices have been designed for performing continuous-flow biochemical and cell-based assays. These devices provide orders of-magnitude reduction in reagent consumption, and offer the potential for implementing high-throughput screening in formats that integrate up-front compound handling with unique assay functionality. PMID- 10679350 TI - Biological function made crystal clear - annotation of hypothetical proteins via structural genomics. AB - Many of the gene products of completely sequenced organisms are 'hypothetical' - they cannot be related to any previously characterized proteins - and so are of completely unknown function. Structural studies provide one means of obtaining functional information in these cases. A 'structural genomics' project has been initiated aimed at determining the structures of 50 hypothetical proteins from Haemophilus influenzae to gain an understanding of their function. Each stage of the project - target selection, protein production, crystallization, structure determination, and structure analysis - makes use of recent advances to streamline procedures. Early results from this and similar projects are encouraging in that some level of functional understanding can be deduced from experimentally solved structures. PMID- 10679351 TI - gamma-tubulin complexes: binding to the centrosome, regulation and microtubule nucleation. AB - Microtubule assembly is initiated in vivo by gamma-tubulin complexes. Cytoplasmic gamma-tubulin complexes are targeted to centrosomes or to other microtubule organizing centers (MTOCs) via a set of so called gamma-tubulin complex binding proteins (GTBPs) that probably interact with the conserved Spc97p/Spc98p protein family of gamma-tubulin complexes. In other cell types, gamma-tubulin complexes may initiate microtubule formation near chromosomes in a MTOC-independent manner. Recently, major advances have been achieved through the finding that gamma tubulin, Spc97p and Spc98p form a conserved core that is probably responsible for microtubule nucleation, and by the discovery that a yeast GTBP is regulated in a cell-cycle-dependent manner and in response to an external signal. Furthermore, it was found that the small GTPase Ran in its GDP-bound state may promote spindle assembly. In addition, an essential function of gamma-tubulin in basal body duplication has been demonstrated in Paramecium. PMID- 10679352 TI - Membrane trafficking, organelle transport, and the cytoskeleton. AB - Cytoskeleton-associated motor proteins typically drive organelle movements in eukaryotic cells in a manner that is tightly regulated, both spatially and temporally. In the past year, a novel organelle transport mechanism utilizing actin polymerization was described. Important advances were also made in the assignment of functions to several new motors and in our understanding of how motor proteins are regulated during organelle transport. In addition, insights were gained into how and why organelles are transported cooperatively along the microtubule and actin cytoskeletons, and into the importance of motor-mediated transport in the organization of the cytoskeleton itself. PMID- 10679354 TI - Microtubule dynamics and tubulin interacting proteins. AB - Microtubule dynamics are crucial in generation of the mitotic spindle. During the transition from interphase to mitosis, there is an increase in the frequency of microtubule catastrophes. Recent work has identified two proteins, Op 18/stathmin and XKCM1, which can promote microtubule catastrophes in vitro and in cells or extracts. Although both of these proteins share the ability to bind tubulin dimers, their mechanisms of action in destabilizing microtubules are distinct. PMID- 10679353 TI - Parallel actin bundles and their multiple actin-bundling proteins. AB - Parallel actin bundles are present in a diverse array of structures, where they are critical determinants of cellular shape and physiology. In the past 18 months, new findings have solidified the concept that parallel actin bundles are assembled in cells through the sequential action of multiple actin-bundling proteins and have begun to shed light on the roles played by the individual actin bundling proteins. PMID- 10679355 TI - Kinesins and microtubules: their structures and motor mechanisms. AB - Atomic resolution three-dimensional structures of two oppositely directed kinesin motors - conventional kinesin and non-claret disjunctional (ncd) protein - are now available in their functional dimeric form. A detailed model of the microtubule has also been recently obtained by docking the 3.7 A structure of tubulin into a 20 A map of the microtubule. Recent structural studies of kinesin motors and their microtubule tracks are contributing to our current understanding of kinesin motor mechanisms. PMID- 10679356 TI - Cytokinesis without myosin II. AB - The ability of substrate-anchored Dictyostelium cells to divide without myosin II has opened the possibility of analysing the formation of cleavage furrows in the absence of a contractile ring made of filamentous myosin and actin. Similar possibilities exist in mutants of budding yeast and, less strictly, also in drug treated mammalian cells. Myosin-II-independent activities in Dictyostelium include the microtubule-induced programming of the cell surface into ruffling areas and regions that are converted into a concave furrow, as well as the translocation of cortexillins and cross-linked membrane proteins towards the cleavage furrow. A centripetal flow of actin filaments followed by their disassembly in the cleavage furrow is proposed to underlie the translocation. PMID- 10679357 TI - Functional cooperation between the microtubule and actin cytoskeletons. AB - In diverse cell types, microtubule (MT) and actin filament networks cooperate functionally during a wide variety of processes, including vesicle and organelle transport, cleavage furrow placement, directed cell migration, spindle rotation, and nuclear migration. The mechanisms by which MTs and actin filaments cooperate to mediate these different processes can be grouped into two broad categories: coordinated MT- and actin-based transport to move vesicles, organelles, and cell fate determinants; and targeting and capture of MT ends at cortical actin sites. Over the past several years, a growing number of cellular factors that bridge these cytoskeletal systems have been identified. These include 'hetero-motor' complexes (physically associated myosin and kinesin), myosin-CLIP170 complexes, formin homology (FH) proteins, dynein and the dynactin complex, Kar9p, coronin, Kelch repeat-containing proteins, and ERM proteins. PMID- 10679358 TI - Control of actin assembly and disassembly at filament ends. AB - The most important discovery in the field is that the Arp2/3 complex nucleates assembly of actin filaments with free barbed ends. Arp2/3 also binds the sides of actin filaments to create a branched network. Arp2/3's nucleation activity is stimulated by WASP family proteins, some of which mediate signaling from small G proteins. Listeria movement caused by actin polymerization can be reconstituted in vitro using purified proteins: Arp2/3 complex, capping protein, actin depolymerizing factor/cofilin, and actin. actin depolymerizing factor/cofilin increases the rate at which actin subunits leave pointed ends, and capping protein caps barbed ends. PMID- 10679359 TI - Functions of unconventional myosins. AB - To date, fourteen classes of unconventional myosins have been identified. Recent reports have implicated a number of these myosins in organelle transport, and in the formation, maintenance and/or dynamics of actin-rich structures involved in a variety of cellular processes including endocytosis, cell migration, and sensory transduction. Characterizations of organelle dynamics in pigment cells and neurons have further defined the contributions made by unconventional myosins and microtubule motors to the transport and distribution of organelles. Several studies have provided evidence of complexes through which cooperative organelle transport may be coordinated. Finally, the myosin superfamily has been shown to contain at least one processive motor and one backwards motor. PMID- 10679360 TI - Intermediate filaments and their associates: multi-talented structural elements specifying cytoarchitecture and cytodynamics. AB - The assembly of intermediate filament (IF) arrays involves the recruitment of a complex set of cell-type-specific IF-associated proteins. Some of them are integral membrane proteins, others act as crosslinking proteins with vectorial binding activities, and yet others comprise motor proteins. In vivo IFs appear to be predominantly heteropolymers, although in vitro several IF proteins (e.g. vimentin, desmin, neurofilament (NF)-L and the nuclear lamins) do self-assemble into IF-like polymers. In contrast, NF-M, NF-H, nestin, synemin and paranemin, all bona fide IF proteins, are unable to self-assemble into IFs either in vitro or in vivo. The individual IF proteins of this large multigene family are chemically heterogeneous, exhibiting different assembly kinetics and yielding discrete types of filaments. The unique physical properties and interaction capabilities of these distinct IF molecular building blocks, in combination with accessory proteins, mediate the generation of a highly dynamic and interconnected, cell-type-specific cytoarchitecture. PMID- 10679361 TI - Focal adhesions - the cytoskeletal connection. AB - Cellular contacts with the extracellular matrix are regulated by the Rho family of GTPases through their effects on both the actin and the microtubule cytoarchitecture. Recent genetic, biochemical and structural data have highlighted the role played by a subset of actin-binding proteins in coupling integrins to cytoskeletal actin and in assembling signalling complexes that are important for cell motility and cell proliferation. PMID- 10679362 TI - How WASP-family proteins and the Arp2/3 complex convert intracellular signals into cytoskeletal structures. AB - In most cells, the structure of the actin cytoskeleton is regulated by Rho-family G proteins. Recent work has outlined a highly conserved signaling pathway from G protein activation to actin assembly. The key downstream components are WASP family proteins - adaptor molecules that bind multiple signaling and cytoskeletal proteins - and the Arp2/3 complex - a multi-functional protein complex that nucleates and crosslinks actin filaments. PMID- 10679363 TI - Actomyosin: law and order in motility. AB - The crystal structures of smooth muscle and scallop striated muscle myosin have both been completed in the past 18 months. Structural studies of unconventional myosins, in particular the stunning discovery that myosin VI moves backwards on actin, are starting to have deep impact on the field and have induced new ways of thinking about actin-based motility. Sophisticated genetic, biochemical and biophysical studies were used to test and refine hypotheses of the molecular mechanism of motility that were developed in the past. Although all these studies confirmed some aspects of these hypotheses, they also raised many new unresolved questions. Much of the evidence points to the importance of the actin-myosin binding process and an associated disorder-to-order transition. PMID- 10679364 TI - How centrioles work: lessons from green yeast. AB - Centrioles are the organizing centers around which centrosomes assemble. Despite a century of study, the molecular details of centriole function and assembly remain largely unknown. Recent work has exploited the unique advantages of unicellular algae to reveal proteins that play central roles in centriole biology. PMID- 10679365 TI - Single molecule analysis of the actomyosin motor. AB - Progress in imaging techniques and nano-manipulation of single molecules has been remarkable. These techniques have allowed the accurate determination of myosin head-induced displacements and of how the mechanical cycles of the actomyosin motor are coupled to ATP hydrolysis. This has been achieved by measuring mechanical and chemical events of actomyosin directly at the single molecule level. Recent studies have made detailed measurements of myosin step size and mechanochemical coupling. The results of these studies suggest a new model for the mechanism of motion underlying actomyosin motors, which differs from the currently accepted lever-arm swinging model. PMID- 10679366 TI - Actin machinery: pushing the envelope. AB - The reconstitution of microbial rocketing motility in vitro with purified proteins has recently established definitively that no myosin motor is required for protrusion. Instead, actin polymerization, in conjunction with a small number of proteins, is sufficient. A dendritic pattern of nucleation controlled by the Arp2/3 complex provides an efficient pushing force for lamellipodial motility. PMID- 10679367 TI - Oxynitrilases for asymmetric C-C bond formation. AB - Oxynitrilases for the preparation of (R)- or (S)-cyanohydrins are now readily available. The research efforts of a number of groups have established these enzymes as catalysts with significant potential for application to asymmetric synthesis. Advances made in molecular cloning and genetics have delivered information on the oxynitrilase mechanism of action and sufficient quantities of enzyme to satisfy industrial requirements. PMID- 10679368 TI - Elucidation of protein-protein interactions using chemical cross-linking or label transfer techniques. AB - Understanding the architectures of multiprotein complexes is a central problem in biology. Of the many chemical methods available, label transfer and cross-linking are becoming more popular. Recently, label transfer has been applied to very large protein complexes with great success, and new oxidative methods for protein cross-linking have been developed that are fast and highly efficient. Advances in these techniques should increase the understanding of biological structures and mechanisms. PMID- 10679369 TI - Enzyme-catalyzed synthesis of carbohydrates. AB - Several new enzymes of utility in the synthesis of carbohydrates have been reported during the past year. Additionally, the utility of several well studied enzymes has been expanded. Pyruvate aldolases, aldolase abzymes and both wild type and mutated glycosidases have found increasing acceptance in the community. Preliminary reports suggest that thermophilic enzymes may possess significant advantages compared to their mesophilic counterparts for carbohydrate synthesis. PMID- 10679370 TI - Nitrile hydrolases. AB - A number of nitrile-related enzymes have been screened over the past year for use in synthetic applications. There have also been significant advances in our understanding of the structures and modes of regulation of metal-containing nitrile hydratases. Enzyme structural characterization has provided new insights into how the molecular structure determines the enzyme function and how an enzyme can be endowed with new properties. This information has important implications for potential future applications other than the present industrial production of acrylamide and nicotinamide. PMID- 10679371 TI - Covalent modification as a strategy to block protein-protein interactions with small-molecule drugs. AB - It is generally difficult to block protein-protein interactions with small molecule drugs. A novel pharmaceutical development strategy to block protein interactions is emerging: targeted covalent modification to sterically block interactions. By this approach, compounds first interact non-covalently with a specific target protein. This interaction juxtaposes a weakly reactive group of the drug with a target amino acid sidechain, which then react by virtue of their high local concentration. PMID- 10679372 TI - Design of novel sequence-specific DNA-binding proteins. AB - The design and selection of DNA-binding proteins or individual domains capable of novel sequence recognition continues to make great strides. Recent studies have also highlighted the role of the non-DNA-contacting portions of the protein and the optimal assembly of the domains. For the first time, it appears that it is possible to produce proteins capable of targeting any gene with an 18 base pair recognition domain. A variety of applications are being explored, such as targeted transcriptional regulation, recombination and viral integration. These proteins will probably find diverse applications in gene therapy, functional genomics, and agriculture. PMID- 10679373 TI - Directed evolution of enantioselective enzymes for organic chemistry. AB - The production of enantiopure compounds is of steadily increasing importance to the chemical and biotechnological industries. In principal, the application of directed evolution in combination with newly developed screening methods enables the generation of enzymes with improved enantioselectivity. The first and most advanced example relates to a bacterial lipase from Pseudomonas aeruginosa. This enzyme was evolved towards a model substrate to yield in a lipase mutant showing > 90% enantiomeric excess as compared to 2% for the wild-type lipase. The creation of enantioselective enzymes by directed evolution will become an important technology in the near future. PMID- 10679374 TI - Small-molecule inhibitors of the cell cycle. AB - The cell cycle remains an attractive target for the development of small-molecule inhibitors for use as both novel chemotherapeutics and research probes. Given the importance of cytoskeletal dynamics and cyclin-dependent kinases for cell-cycle progression, much interest has focused on the identification of anti-mitotic agents and kinase inhibitors. However recent advances in cell-based screening technologies and an increased interest in inhibitors with greater specificity are beginning to influence the search for novel cell-cycle inhibitors. PMID- 10679375 TI - Role of linkers in communication between protein modules. AB - Multidomain proteins are common in a variety of cellular processes. Their domains are interconnected through short stretches of amino acid residues referred to as linkers. Recent studies on many systems have provided compelling evidence that linkers are more than simple covalent connectors. They also perform the important task of establishing communication between the different functional modules that exist within such proteins. PMID- 10679376 TI - Modifying ligand specificity of gene regulatory proteins. AB - Nuclear receptors contain a conserved hydrophobic ligand binding pocket that is particularly amenable to structure-based protein engineering. Thus, site-directed mutagenesis of the ligand binding pocket has resulted in the creation of nuclear receptors with novel ligand specificities. Such proteins are now being used to control gene expression in vivo in a ligand-dependent manner. PMID- 10679377 TI - Combinatorial protein reagents to manipulate protein function. AB - The design and use of combinatorial protein libraries has become a fast moving field in molecular biology. Different experimental systems supporting various selection schemes are now available. The latest breakthroughs include evolutionary experiments to improve existing binding surfaces, selections of homodimerizing peptides, the use of peptide aptamers to disrupt protein interactions inside living cells, and functional selections of aptamers to probe regulatory networks. PMID- 10679378 TI - Engineering biosynthetic pathways: new routes to chiral amino acids. AB - The engineering of microbial biosynthetic pathways is advancing rapidly because of new molecular genetic approaches, sophisticated analysis of metabolic flux and the rapid sequencing of diverse bacterial genomes. The classical methods of mutagenesis/selection, originally applied in the development of amino-acid-over producing bacteria are now being complemented by an increasingly rational strategy. PMID- 10679379 TI - Biocatalysis in low-water media: understanding effects of reaction conditions. AB - The key role played by counter-ions with enzymes in low-water systems has become better appreciated with, for example, large effects on enantioselectivity. In low dielectric media, counter-ions will associate strongly with charges in the protein or its substrates. Studies of temperature dependence have shown that hard to-model entropies have a significant effect on behaviour, including enantioselectivity. Evidence has been presented that the supramolecular organisation of enzyme molecules can have important effects on behaviour, for example collapse of microstructure in cross-linked crystals. PMID- 10679380 TI - New approaches for cell-specific targeting: identification of cell-selective peptides from combinatorial libraries. AB - Peptides that recognize specific cell types promise to be valuable tools both in research and clinical applications. Cell-specific peptides can be useful as drug delivery vehicles, diagnostic agents, affinity reagents for cell purification, gene therapy delivery agents, and research tools to probe the nature of a cell's surface. Recently, cell-specific targeting-peptides have been identified by phage display selections against purified cell-surface markers, whole cells in tissue culture, and even tissues within live animals. These methods for identifying cell targeting peptides will certainly increase the tools available to the scientist for cell-specific targeting. PMID- 10679381 TI - Observation of unstable species in enzyme-catalyzed transformations using protein crystallography. AB - Recent advances in rapid X-ray diffraction data collection methods, cryocrystallography, and other techniques have made it possible to visualize short-lived species in enzyme-catalyzed reactions directly at atomic resolution for a significant number of crystalline enzymes. The wide range of reaction types, intermediate lifetimes, and crystal characteristics means that different methods must be employed in each case, but there are enough examples now of successful structure determinations of normally unstable species to suggest guidelines for future investigations. PMID- 10679382 TI - Molecular modeling and biocatalysis: explanations, predictions, limitations, and opportunities. AB - Rapid advances in structural biology have revealed the three-dimensional structures of many biocatalysts. Molecular modeling is the tool that links these structures with experimental observations. As a qualitative tool, current modeling methods are extremely useful. They can explain, on a molecular level, unusual features of reactions. They can predict how to increase the selectivity either by substrate modification or by site-directed mutagenesis. Quantitative predictions, for example the degree of enantioselectivity, are still not reliable, however. Modeling is limited also by the availability of three dimensional structures. Most current modeling involves hydrolases, especially proteases and lipases, but structures for other types of enzymes are starting to appear. PMID- 10679383 TI - The ARF/p53 pathway. AB - The ARF tumor suppressor connects pathways regulated by the retinoblastoma protein and p53. ARF inactivation reduces p53-dependent apoptosis induced by oncogenic signals. Nucleolar relocalization of Mdm2 by ARF connotes a novel mechanism for preventing p53 turnover and provides a framework for understanding how stress signals cooperate to regulate p53 function. PMID- 10679384 TI - RecQ family helicases: roles in cancer and aging. AB - The RecQ family of DNA helicases includes at least three members in humans that are defective in genetic disorders associated with cancer predisposition and/or premature aging. Recent studies have shed light on the roles of RecQ helicases in suppressing 'promiscuous' genetic recombination and in ensuring accurate chromosome segregation. In particular, the biochemical properties of several family members have been characterised and functional interactions with other nuclear proteins have been defined. PMID- 10679385 TI - Complexity in the spindle checkpoint. AB - Cell viability requires accurate chromosome segregation at mitosis. The spindle checkpoint ensures that anaphase is not attempted until the sister chromatids of each chromosome are attached to spindle microtubules from opposite poles. The checkpoint mechanism involves a signal transduction cascade that is more complex than was originally envisioned. PMID- 10679386 TI - Tumor suppressor genes. AB - Although tumor suppressor genes continue to be discovered, the most recent advances have been made in attributing new and exciting functions to existing ones - such as the apparent role of VHL as a regulator of proteolysis. Great insights have also come from piecing genes together into pathways and networks. For instance the discovery that cyclin D1 is regulated by beta-catenin/Tcf-4 allows us to tie the APC pathway to the RB pathway and cell cycle control. Similarly, tumor suppressor genes have been fitted together with oncogenes into the various pathways that regulate apoptosis such that tumor suppressor function is now attributed to some of the basic components of the apoptotic machinery, such as caspases and Apaf-1. The great pace at which mouse models of tumorigenesis continue to advance our knowledge of tumor suppressor gene function has led us to look anew at the role of genes such as TCF-1 and SMAD-3 in human cancer. Finally, the realisation that different growth regulatory pathways give rise to generic signals suggests that future work may lie in integrating the signals from different pathways and in understanding the importance of protein levels to cellular function. PMID- 10679387 TI - The genetic control of organ growth: insights from Drosophila. AB - The genetic control of growth ensures that animals grow to reproducible sizes and that tumourous growth is rare. This year, the regulation of organ growth has been studied extensively in Drosophila imaginal discs, and a signalling pathway that regulates organ growth and size has been identified. Furthermore, the role of Drosophila homologues to human tumour suppressor genes and oncogenes in imaginal disc growth has been investigated. PMID- 10679388 TI - Extrinsic regulators of epithelial tumor progression: metalloproteinases. AB - Extracellular metal-dependent proteinases regulate cell behavior by remodeling stromal and cell surface proteins, thereby influencing cell recruitment, cell shape, motility, proliferation, survival, genomic (in)stability, and differentiation. In recent years, the importance of proteinase-induced signaling has been underscored by evidence that altered regulation of cell-extracellular matrix and cell-cell interactions by proteinases can contribute, in a causal manner, to neoplastic progression. PMID- 10679389 TI - Controlling the end of the cell cycle. AB - The past year has seen significant advances in our understanding of how the events which occur at the end of mitosis, such as cytokinesis and the inactivation of mitotic cyclin dependent kinases are triggered, and also how they are prevented from occurring prematurely or inappropriately. This control is achieved through a combination of temporally ordered proteolytic events and changes in the subcellular localisation of proteins. These studies have also revealed that the nucleolus and spindle pole bodies play a key role in this regulation. PMID- 10679390 TI - Viral-encoded cyclins. AB - D-type cyclin homologs have been found in the genomes of herpesviruses associated with neoplasias. They appear to exploit features of G(1) cyclins but extend their properties to allow for deregulation of the cell cycle. Advances in the study of the molecular basis for these novel features as well as the potential role of viral cyclins in tumorigenesis are addressed. PMID- 10679391 TI - Action of Myc in vivo - proliferation and apoptosis. AB - The protein products of many dominant oncogenes are capable of inducing both cell proliferation and apoptosis. Recent experiments employing transgenic mice that express an ectopically regulatable myc gene or protein have begun to elucidate the role of the balance between proliferation and apoptosis in Myc-induced carcinogenesis. An outstanding feature of these experiments is the demonstration that the balance between oncogene-induced proliferation and apoptosis in a given tissue can be a critical determinant in the initiation and maintenance of the tumor. PMID- 10679392 TI - A critical role for telomeres in suppressing and facilitating carcinogenesis. AB - Progressive telomere shortening occurs with the division of primary human cells and activates tumor suppressor pathways, triggering senescence and inhibiting tumorigenesis. Loss of p53 function, however, allows continued cell division despite increasing telomere dysfunction and entry into telomere crisis. Recent data suggest that the severe chromosomal instability of telomere crisis promotes secondary genetic changes that facilitate carcinogenesis. Reactivation of telomerase stabilizes telomere ends and allows continued tumor growth. PMID- 10679393 TI - Cloak and dagger in the avoidance of immune surveillance. AB - CD95 and CD95-ligand (CD95L) are physiological mediators of apoptosis required for the control of cell numbers in the human immune system. Discoveries in CD95 dependent mechanisms of immune evasion by tumours suggest regulation by oncogene expression. Clonal contraction of lymphocytes by a CD95/CD95L-independent mechanism has been reported and new evidence supports a role for CD95-dependent peripheral lymphocyte deletion by non-lymphoid tissue. Additionally, factors affecting the pro- and anti-inflammatory effects of CD95L point to a balance of cytokines and growth factors. PMID- 10679394 TI - Proteolysis and the cell cycle: with this RING I do thee destroy. AB - The ubiquitin system drives the cell division cycle by the timely destruction of numerous regulatory proteins. Remarkably, the two main activities that catalyze substrate ubiquitination in the cell cycle, the Skp1-Cdc53/cullin-F-box protein (SCF) complexes and the anaphase-promoting complex/cyclosome (APC/C), define a new superfamily of E3 ubiquitin ligases, all based on related cullin and RING-H2 finger protein subunits. The circuits that interconnect the SCF, APC/C and cyclin dependent kinase activities form a master oscillator that coordinates the replication and segregation of the genome. PMID- 10679395 TI - Sensing and responding to DNA damage. AB - DNA damage or stalled DNA replication can activate specific signal transduction pathways, termed checkpoints. Checkpoint activation can result in increased repair, induction of a transcriptional programme and inhibition of cell-cycle progression. Recent results have suggested possible mechanisms for the detection of specific DNA structures, provided further information on the organisation of the signal transduction cascade and demonstrated involvement of the checkpoint pathway in DNA repair. PMID- 10679396 TI - Cell-cycle checkpoints that ensure coordination between nuclear and cytoplasmic events in Saccharomyces cerevisiae. AB - Cytoskeletal organization is crucial for several aspects of cell-cycle progression but cytoskeletal elements are quite sensitive to environmental perturbations. Two novel checkpoint controls monitor the function of the actin and microtubule systems in budding yeast and operate to delay cell-cycle progression in response to cytoskeletal perturbations. In cells whose actin cytoskeleton has been perturbed, bud formation is frequently delayed and the morphogenesis checkpoint introduces a compensatory delay of nuclear division until a bud has been formed. In cells whose microtubule cytoskeleton has been perturbed, anaphase spindle elongation often occurs entirely within the mother cell, and the post-anaphase nuclear migration checkpoint introduces a compensatory delay of cytokinesis until one pole of the anaphase nucleus enters the bud. Recent studies indicate that regulators of entry into mitosis are localized to the daughter side of the mother-bud neck whereas regulators of exit from mitosis are localized to the spindle pole bodies. Thus, specific cell-cycle regulators are well-placed to monitor whether a cell has formed a bud and whether a daughter nucleus has been delivered accurately to the bud following mitosis. PMID- 10679397 TI - Integration of positive and negative growth signals during ras pathway activation in vivo. AB - Expression of RAS proteins can have either positive or negative effects on cell growth, differentiation and death. New technologies are being developed for the generation of animal models to address the questions of where, when and how much Ras is expressed during tumorigenesis, and how these disparate signals are integrated during multistage carcinogenesis. PMID- 10679398 TI - The role of IL-18 in innate immunity. AB - IL-18 - a novel cytokine with potent IFN-gamma-inducing activities - plays an important role in the Th1-mediated immune response in collaboration with IL-12. IL-18 is a member of the IL-1 family. The IL-18 receptor system and its signal transduction pathway are analogous to those of the IL-1 receptor. Mice deficient in IL-18 have demonstrated its critical role in natural killer cell activity and in vivo Th1 response. PMID- 10679399 TI - NF-kappaB and the innate immune response. AB - In the innate immune reaction, microbial pathogens activate phylogenetically conserved cellular signal transduction pathways that regulate the ubiquitous nuclear factor-kappaB (NFkappaB). NF-kappaB has pleiotropic functions in immunity; however, it is also critical for development and cellular survival. Many aspects of how the different pathways utilize a common kinase complex that ultimately activates NF-kappaB have been clarified by gene inactivation and biochemical analysis. PMID- 10679400 TI - CD14-dependent clearance of apoptotic cells: relevance to the immune system. AB - Until very recently, the function of CD14 was thought to be limited to innate immune responses to bacterial and other microbial structures. It is now known that macrophage CD14 serves an additional unexpected function, namely as a receptor involved in the recognition and phagocytosis of cells undergoing apoptosis. In stark contrast to its role in eliciting pro-inflammatory responses following binding of microbial ligands, macrophage CD14 mediates clearance of apoptotic cells without inciting inflammation. Increasing interest in the profoundly important final stage of apoptosis - the engulfment process - together with significant advances in knowledge of the immunological consequences of apoptotic-cell clearance and of the means by which signal transduction may be achieved following CD14-ligand binding have begun to produce a clearer picture of the role of CD14 in the immune system. PMID- 10679401 TI - Antigen arrays in T cell immunology. AB - The screening of compound arrays in in vitro bioassays has developed into a powerful tool for the identification of biologically active substances. In the past decade, this technology has increasingly been applied to immunology. As the specificity of the immune system is determined by antigen detection via receptors on B and T cells, targeting the specificity of these immune receptors with random arrays is unique in its ability to generate general and quantitative information on cellular (cross-)reactivity. Synthetic array studies have been useful for identification of epitopes and antigens from databases by defining recognition patterns, isolation of synthetic peptides capable of modulating T cell responsiveness, characterisation of TCR promiscuity, and identification of functionally cross-reacting peptides that are potentially involved in molecular mimicry. PMID- 10679402 TI - Accessory molecules for MHC class II peptide loading. AB - Accessory molecules, such as HLA-DM and invariant chain, modulate the ligands bound to MHC class II molecules in antigen-presenting cells. Recent investigations, including gene targeting experiments, have shed light on the functions of these molecules, their mechanisms of action, interactions with class II molecules, and the relationships with associated molecules such as tetraspanins and HLA-DO. PMID- 10679403 TI - Human complement regulators: a major target for pathogenic microorganisms. AB - The C3 convertases of the human complement system are controlled by fluid-phase and membrane proteins in the RCA (regulators of complement activation) family. Accumulated data show that many pathogenic microorganisms interact with these complement regulators. Recent advances in this field include determination of the crystal structure of the binding domains in the measles virus receptor CD46 and identification of a CD46 transgenic mouse line that is sensitive to measles virus. Moreover, recent findings support the hypothesis that pathogenic bacteria binding fluid-phase RCA proteins exploit these proteins to escape complement attack. These studies provide novel insight into the interplay between pathogens and the innate immune system and may have implications for the plans to use animals expressing an RCA protein for xenotransplantation. PMID- 10679404 TI - Reactive oxygen and reactive nitrogen intermediates in innate and specific immunity. AB - Nitric oxide, nitric oxide derivatives and reactive oxygen intermediates are toxic molecules of the immune system which contribute to the control of microbial pathogens and tumors. There is recent evidence for additional functions of these oxygen metabolites in innate and adaptive immunity; these functions include the modulation of the cytokine response of lymphocytes and the regulation of immune cell apoptosis, as well as immunodeviating effects. Components of several signal transduction pathways have been identified as intracellular targets for reactive nitrogen and oxygen intermediates. PMID- 10679405 TI - Rational antigen modification as a strategy to upregulate or downregulate antigen recognition. AB - Recent and rapid advances in our understanding of the cellular and molecular mechanisms of antigen recognition by CD8(+) and CD4(+) T lymphocytes have led to the birth of possibilities for site-directed, rational modification of cognate antigenic determinants. This immunologic concept has vast biomedical implications for regulation of host immunity against the pathogenesis of diverse disease processes. The upregulation of antigen-specific T-cell responses by 'agonistic' peptides would be most desirable in response to invasive pathogenic challenges, such as infectious and neoplastic disease, while the downregulation of antigen specific T-cell responses by 'antagonistic' peptides would be most efficacious during inappropriate pathologic consequences, such as autoimmunity. The capacity to experimentally manipulate intrinsic properties of cognate peptide ligands to appropriately alter the nature, course and potency of cellular immune interactions has important potential in both preventive and therapeutic clinical paradigms. PMID- 10679406 TI - The role of CpG motifs in innate immunity. AB - Pattern recognition receptors of the innate immune system are able to distinguish certain prokaryotic DNAs from vertebrate DNAs by detecting unmethylated CpG dinucleotides in particular base contexts ('CpG motifs'). Recent studies have begun to define the molecular mechanisms of actions of CpG motifs and have demonstrated their stimulatory effects on leukocytes from humans and vertebrates other than mice. Oligodeoxynucleotides containing CpG motifs are highly effective Th1-like vaccine adjuvants through multiple routes of immunization and show promise as immunotherapeutic agents for cancer and allergic diseases. PMID- 10679407 TI - Toll signaling pathways in the innate immune response. AB - The Toll signaling pathway, which is required for the establishment of the dorsal ventral axis in Drosophila embryos, plays an important role in the response of larval and adult Drosophila to microbial infections. Recent genetic evidence has shown that a mammalian Toll-like receptor, mouse Tlr4, is the signal transducing receptor activated by bacterial lipopolysaccharide. Thus, Toll-like receptors appear to detect a variety of microbial components and to trigger a defensive reaction in both Drosophila and mammals. Genetic data from both Drosophila and mice have defined components required for activation of Toll-like receptors and for the downstream pathways activated by the Toll-like receptors. PMID- 10679408 TI - The control of T cell responses by dendritic cell subsets. AB - Dendritic cells are known as the most efficient antigen-presenting cell type to activate naive T cells; however, they are able to do more than just efficiently present antigen to T cells. They are key modulators of the immune response that can influence Th cell differentiation by preferentially inducing Th type 1 or 2 cell responses, and the differential polarisation of CD4(+) T cells appears to be mediated by discrete dendritic cell subsets. PMID- 10679409 TI - Cathepsins and compartmentalization in antigen presentation. AB - Intracellular trafficking and cell surface expression of MHC class II molecules is a tightly regulated process and is to a large extent, determined by the fate of the class II chaperone, the invariant chain. Inhibition of endosomal proteases critical to invariant chain proteolysis reveals marked shunting of class II complexes to lysosomal compartments. Regulation of endosomal protease activity by expression of cystatin C directs class II cell surface expression during maturation of dendritic cells. These studies highlight the taut interactions between class-II-invariant-chain complexes and endosomal proteases during MHC class II maturation. PMID- 10679410 TI - Identification of HLA-class-II-restricted epitopes of autoantigens in transgenic mice. AB - The past year has seen a spate of research in the use of HLA transgenic mice in the identification of self antigens associated with autoimmunity. Dominant T cell determinants - and, in a few cases, B cell determinants - have been characterized in the context of disease-predisposing HLA DR and DQ genes for at least three prominent and devastating autoimmune diseases: rheumatoid arthritis, multiple sclerosis and insulin-dependent diabetes mellitus. PMID- 10679411 TI - Tlr4: central component of the sole mammalian LPS sensor. AB - Mutations of the mouse Lps locus abolish responses to lipopolysaccharide (LPS). Positional cloning work has revealed that Lps encodes the Toll-like receptor 4 (Tlr4), which functions as the transmembrane component of the LPS receptor complex, an unduplicated pathway for the detection of endotoxin. The structurally related protein Tlr2 makes no contribution to LPS signal transduction. PMID- 10679412 TI - From synapses to immunological memory: the role of sustained T cell stimulation. AB - T cell activation is a sustained process driven by antigen and cytokines, which results in the generation of large numbers of effector and memory cells. Recent experiments from different fields have shed light on the mechanisms that maintain the signaling process at the level of a single synapse between a T cell and an antigen-presenting cell, as well as at the level of a secondary lymphoid organ, in the course of the immune response. These findings explain the unique capacity of the immune system to discriminate between antigens from infectious and noninfectious agents. PMID- 10679413 TI - Cellular routes of invasion by enteropathogens. AB - The cellular pathways of infection utilized by pathogenic enteric bacteria have important implications for their clinical manifestations. Yersinia reaches Peyer's patches via M cells and uses plasmid-encoded factors to resist phagocytic cells. Shigella also translocates via M cells and incapacitates phagocytes, but subsequently re-enters the epithelium basolaterally to elicit an acute inflammatory response. Salmonella has recently been shown to both colonize Peyer's patches via M cells and independently disseminate to extraintestinal sites via CD18-expressing phagocytes. M cell-mediated entry can lead to gastroenteritis and mucosal antibody production, while systemic dissemination can result in septicemia and elicitation of systemic immune responses. PMID- 10679414 TI - Novel approaches to monitor bacterial gene expression in infected tissue and host. AB - Elucidating the complex and dynamic host-microbe interactions during infection requires, among other things, detailed knowledge of microbial gene expression in vivo. Recently, advances in fluorescence and bioluminescence detection techniques, as well as recombinase-based in vivo expression technology, have rendered monitoring virulence gene expression in vivo a feasible task. These techniques have been adapted by several laboratories to study the spatial and temporal patterns of virulence gene expression by organisms such as Salmonella typhimurium, Listeria monocytogenes, Yersinia entercolitica and Vibrio cholerae during infection of tissue culture or animal models of infection. PMID- 10679415 TI - Caenorhabditis elegans: a model genetic host to study Pseudomonas aeruginosa pathogenesis. AB - In the past year, a Caenorhabditis elegans-Pseudomonas aeruginosa pathogenesis model has been developed to facilitate the systematic dissection of both host and pathogen genes involved in pathogenic interactions. Analysis of the P. aeruginosa C. elegans interaction should shed light on the larger question of how organisms interact at the molecular level in antagonistic relationships. PMID- 10679416 TI - Analyzing the molecular foundations of commensalism in the mouse intestine. AB - We maintain complex societies of nonpathogenic microbes on our mucosal surfaces. Although the stability of this flora is important for human health, very little is known about how its constituents communicate with us to forge stable and mutually advantageous relationships. The vast majority of these indigenous microbes reside in the intestine. Recent studies of a gut commensal, Bacteroides thetaiotaomicron, has revealed a novel signaling pathway that allows the microbe and host to actively collaborate to produce a nutrient foundation that can be used by this bacterium. This pathway illustrates the type of dynamic molecular interactions that help define commensal relationships. PMID- 10679417 TI - Reactive nitrogen intermediates and the pathogenesis of Salmonella and mycobacteria. AB - Over the past decade, reactive nitrogen intermediates joined reactive oxygen intermediates as a biochemically parallel and functionally non-redundant pathway for mammalian host resistance to many microbial pathogens. The past year has brought a new appreciation that these two pathways are partially redundant, such that each can compensate in part for the absence of the other. In combination, their importance to defense of the murine host is greater than previously appreciated. In addition to direct microbicidal actions, reactive nitrogen intermediates have immunoregulatory effects relevant to the control of infection. Genes have been characterized in Mycobacterium tuberculosis and Salmonella typhimurium that may regulate the ability of pathogens to resist reactive nitrogen and oxygen intermediates produced by activated macrophages. PMID- 10679418 TI - Genetic susceptibility to intracellular infections: Nramp1, macrophage function and divalent cations transport. AB - Nramp1 is one of the few host resistance genes that have been characterized at the molecular level. Nramp1 is an integral membrane protein expressed in the lysosomal compartment of macrophages and is recruited to the membrane of bacterial phagosomes where it affects intracellular microbial replication. Nramp1 is part of a very large gene family conserved from bacteria and man that codes for transporters of divalent cations transporters. We propose that Nramp1 affects the intraphagosomal microbial replication by modulating divalent cations content in this organelle. Both mammalian and bacterial transporters may compete for the same substrate in the phagosomal space. PMID- 10679419 TI - Bacterial pili: molecular mechanisms of pathogenesis. AB - Gram-negative bacteria produce a diverse array of pili that mediate microbe microbe and host-pathogen interactions important in the development of disease. The structural and functional characterization of these organelles, particularly their role in triggering signals in both the bacterium and the host upon attachment, has begun to reveal the molecular mechanisms of bacterial diseases. PMID- 10679420 TI - Bacterial replication in the host cell cytosol. AB - Intracellular bacteria in mammalian host cells can either live in a membrane bound vacuole modified to support bacterial growth, or escape from the primary phagosome into the host cell cytoplasm. Phagosomal escape is best studied in Listeria monocytogenes in which a pore-forming cytolysin and two phospholipases are involved in the lysis of the phagosomal membrane. The mechanisms of and requirements for cytoplasmic growth are less clear but there is growing evidence that proficient replication of bacteria in the cytoplasmic compartment requires specific bacterial and cellular preconditions. PMID- 10679421 TI - Effector proteins of phytopathogenic bacteria: bifunctional signals in virulence and host recognition. AB - Phytopathogenic bacteria deliver effectors of disease into plant hosts via a Type III secretion system. These Type III effectors have genetically determined roles in virulence. They also are among the components recognized by the putative receptors of the plant innate immune system. Recent breakthroughs include localization of some of these Type III effectors to specific host cell compartments, and the first dissection of pathogenicity islands that carry them. PMID- 10679422 TI - Bacterial adaptation to host innate immunity responses. AB - Several recent reports show that different bacterial components trigger innate and inflammatory responses in host organisms. In parallel, selected bacterial virulence factors have been identified that interfere with corresponding responses. In many cases, this involves interference with host proinflammatory signal transduction pathways, whereas in selected cases bacterial virulence factors interfere with host antibacterial mechanisms. This indicates that bacteria, besides activating cellular responses, also have the capacity to directly interact with branches of the innate defence. PMID- 10679423 TI - Borrelia pathogenesis research in the post-genomic and post-vaccine era. AB - In the two years after publication of the genome sequence of Borrelia burgdorferi and reports on human field trials of a vaccine against Lyme borreliosis, there has been further progress in understanding of host-parasite interactions during Lyme borreliosis and relapsing fever. Some mechanisms that Borrelia spirochetes use to avoid elimination and to persist in the host are novel. In addition, the recent discovery of antigenic variation in the Lyme disease agent B. burgdorferi adds to the complexity of the possible virulence properties of this human pathogen. PMID- 10679424 TI - Manipulating the host to study bacterial virulence. AB - The ability to manipulate animal hosts as well as bacterial pathogens greatly expands the utility of in vivo models of infection. For example, the construction of mice that harbor human tissues or express specific transgenes can provide ligand-receptor interactions that are essential for pathogenesis. Interactions between virulence factors and specific host defenses can sometimes be resolved by challenging selectively immuno deficient mice with bacteria containing virulence gene mutations. Transgenic animals expressing inducible reporters can be used to conveniently identify cells in which specific response pathways have been activated during infection. These and other approaches promise to improve the quality of information obtainable from in vivo assessments of pathogenesis. PMID- 10679425 TI - Endotoxin, toll-like receptor 4, and the afferent limb of innate immunity. AB - Positional cloning work and subsequent biochemical analyses have revealed that Toll-like receptor 4 (Tlr4) transduces the lipopolysaccharide (LPS) signal, alerting the host to infection by Gram-negative bacteria. Moreover, it appears that the LPS sensing pathway is a solitary one: disruption of Tlr4 causes complete unresponsiveness to LPS. As several Tlr family members exist in vertebrates, it appears likely that the innate immune system defends the host by recognizing a small number of structurally conserved molecules that distinguish the microbial world from tissues of the host. PMID- 10679426 TI - Signaling mechanisms in the antimicrobial host defense of Drosophila. AB - Drosophila has appeared in recent years as a powerful model to study innate immunity. Several papers published in the past year shed light on the role of the three Rel proteins Dorsal, Dif and Relish in the regulation of antimicrobial peptide expression. In addition, the discovery that a blood serine protease inhibitor is involved in the control of the antifungal response indicates that Toll is activated upon triggering of a proteolytic cascade and does not function as a Drosophila pattern recognition receptor. PMID- 10679427 TI - Development of neuron-neuron synapses. AB - Our understanding of neuronal synapse development has advanced in recent years. The development of glycinergic synapses appears to depend on gephyrin and glycine receptor activity. Molecular characterization of the structure and development of glutamatergic synapses is in progress, but the underlying mechanisms remain unclear. Activity-dependent mechanisms and specific molecules that regulate the morphological development of dendritic spines have recently been identified. PMID- 10679428 TI - Critical periods during sensory development. AB - Recent studies have made progress in characterizing the determinants of critical periods for experience-dependent plasticity. They highlight the role of neurotrophins, NMDA receptors and GABAergic inhibition. In particular, genetic manipulation of a single molecule, brain-derived neurotrophic factor (BDNF), has been shown to alter the timing of the critical period of plasticity in mouse visual cortex, establishing a causal relation between neurotrophin action, the development of visual function, and the duration of the critical period. PMID- 10679429 TI - The GDNF family ligands and receptors - implications for neural development. AB - The glial cell line derived neurotrophic factor (GDNF) family has recently been expanded to include four members, and the interactions between these neurotrophic factors and their unique receptor system is now beginning to be understood. Furthermore, analysis of mice lacking the genes for GDNF, neurturin, and their related receptors has confirmed the importance of these factors in neurodevelopment. The results of such analyses reveal numerous similarities and potential overlaps in the way the GDNF and the nerve growth factor (NGF) families regulate development of the peripheral nervous system. PMID- 10679430 TI - Vnd/nkx, ind/gsh, and msh/msx: conserved regulators of dorsoventral neural patterning? AB - Expression of vnd in ventral, ind in intermediate, and msh in dorsal columns of fly neurectoderm, and of homologous gene families in corresponding domains of vertebrate neurectoderm, suggests that elements of dorsoventral neural patterning have been evolutionarily conserved. However, upstream signaling pathways regulating this columnar gene expression pattern appear to have diverged significantly throughout evolution. In addition, while recent loss-of-function studies in flies and mice indicate that these three genes may have a conserved role in regional specification, there is no obvious conservation of the particular cell fates deriving from corresponding domains. The three-column expression pattern may thus represent a developmental mechanism that is more resistant to evolutionary changes than genetic events upstream or downstream of it. PMID- 10679431 TI - Molecular mechanisms for activity-regulated protein synthesis in the synapto dendritic compartment. AB - The creation of enduring modifications in synaptic efficacy requires new protein synthesis. Neurons face the formidable challenge of directing these newly made proteins to the appropriate subset of synapses. One attractive solution to this problem is the local translation of mRNAs that are targeted to dendrites and perhaps to synapses themselves. The molecular mechanisms mediating such local protein synthesis, notably CPEB-mediated cytoplasmic polyadenylation, are now being elucidated. PMID- 10679432 TI - Axonal signals in the assembly of neural circuitry. AB - Recent work in Drosophila and rodents has revealed that proteins transported along axons and delivered to pathway and target cell populations play important roles in the construction of neural circuitry. Interestingly, the parallels between these systems may extend to the identities of some of the molecules involved. PMID- 10679433 TI - Asymmetric division of Drosophila neural stem cells: a basis for neural diversity. AB - Recent studies of Drosophila neural precursor cells have unveiled the essential roles played by asymmetric cell divisions in the determination of cell fates during neural development. Our understanding now extends to the molecular nature of the cell polarity that underlies asymmetric divisions. This polarity is conserved among neural stem cells, epithelial cells and fertilized eggs. PMID- 10679434 TI - Transcriptional mechanisms in the development of motor control. AB - Considerable progress has been made in understanding how combinatorially expressed transcription factors control the development of neuronal subtypes in the fly and vertebrate central nervous systems. The mode of action of many of these factors has been conserved from invertebrates to vertebrates throughout evolution, such as the formation and regulation of specific transcriptional complexes, the utilization of repressors for maintaining specificity, and the use of phosphorylation as an important means for transiently altering transcriptional activity. PMID- 10679435 TI - Notch and presenilins in vertebrates and invertebrates: implications for neuronal development and degeneration. AB - Recent progress in elucidating the biology of Notch and presenilin has revealed a close functional relationship between these two proteins during cell fate determination in worms, flies and humans. Presenilins are required for the putatively intramembranous proteolysis of Notch to release its intracellular domain to the nucleus. This finding establishes a specific biochemical role for presenilins in Notch signaling and interfaces with emerging evidence about how frizzled, disheveled and numerous other genes regulate the highly complex Notch pathway. Advances in understanding Notch and presenilin functions in the differentiation of neurons and non-neural cells have important implications not only for development but also for late-life degenerative disorders such as Alzheimer's disease. PMID- 10679436 TI - Active killing of neurons during development and following stress: a role for p75(NTR) and Fas? AB - Evidence for active triggering of neuronal death continues to accumulate. The transmembrane receptors p75(NTR) and Fas can trigger (and in some cases are required for) programmed cell death of the neurons that express them, through signalling pathways that are regulated by a variety of cytoplasmic effectors. Neuronal death induced by trophic deprivation often requires Fas signalling, further blurring the boundaries between naturally occurring and stress-induced neuronal death. PMID- 10679437 TI - Get to know your stem cells. AB - Our view of the central nervous system has changed dramatically over the past few years. It is now well established that new neurons are generated continuously in adult mammals, including humans. These neurons derive from self-renewing multipotent neural stem cells. The identify of these stem cells has recently been unveiled. PMID- 10679438 TI - Semaphorins and their receptors in vertebrates and invertebrates. AB - The semaphorins are a family of intercellular signaling proteins that has grown to include 19 identified members in higher vertebrates. Several of its members act as axonal guidance molecules. One participates in signaling in the immune system. The majority, however, do not yet have known biological functions. Recent studies have shown that neuropilins and plexins act as receptors for semaphorins. The most important challenge for the future is to define the biological roles of semaphorins in vivo. PMID- 10679439 TI - From Abl to actin: Abl tyrosine kinase and associated proteins in growth cone motility. AB - The Abl tyrosine kinase plays an important role in axonogenesis. Recent reports indicate that this role involves interaction with several different protein families, including LAR phosphatases, catenin/cadherin cell adhesion complexes, Trio family GEFs, and Ena/VASP family actin regulatory proteins. These findings suggest that Abl and its associated proteins may regulate cell adhesion and actin polymerization, thereby regulating growth cone motility during axonogenesis. PMID- 10679440 TI - Rapid dendritic movements during synapse formation and rearrangement. AB - Major technical advances in the imaging of live cells have led to a recent flurry of studies demonstrating how dendrites remodel dynamically during development. Taken together with our current understanding of axonal development, these studies help provide a more unified picture of how neural circuits might be formed altered or maintained throughout life. PMID- 10679441 TI - Discussion point. The case for floor plate induction by the notochord. PMID- 10679442 TI - The broken mouse: the role of development, plasticity and environment in the interpretation of phenotypic changes in knockout mice. AB - With the advent of gene knockout technology has arisen the problem of how to interpret the resulting phenotypic changes in mice lacking specific genes. This problem is especially relevant when applied to behavioral phenotypes of knockout mice, which are difficult to interpret. Of particular interest are the roles of development and compensatory changes, as well as other factors, such as the influence of the gene knockout on nearby genes, the effect of the genetic background strain, maternal behavioral influences, and pleiotrophy. PMID- 10679443 TI - Discussion point. Origin and specification of the neural tube floor plate: insights from the chick and zebrafish. PMID- 10679444 TI - Slit proteins: key regulators of axon guidance, axonal branching, and cell migration. AB - In the past year, Slit proteins have been identified as important regulators of axon guidance and cell migration in Drosophila and vertebrates. Remarkably, they were simultaneously identified as negative regulators, repelling various axonal and cell migrations in both invertebrates and vertebrates, and as positive regulators, stimulating branching and extension of at least one class of axons in vertebrates. PMID- 10679445 TI - Cell signaling within the shoot meristem. AB - Shoot apical meristems are self-renewing stem cell populations that generate all of the above-ground organs (i.e. stems, leaves and flowers) of higher plants. Recent studies have identified new molecular components required for proper shoot meristem activity, and they have revealed that complex, intercellular communication pathways play important roles in coordinating meristem function. PMID- 10679446 TI - How plants tell the time. AB - The components of the circadian system that have recently been discovered in plants share some characteristics with those from cyanobacterial, fungal and animal circadian clocks. Light input signals to the clock are contributed by multiple photoreceptors: some of these have now been shown to function specifically in response to light of defined wavelength and fluence rate. New reports of clock-controlled processes and genes are highlighting the importance of time management for plant development. PMID- 10679447 TI - The YABBY gene family and abaxial cell fate. AB - The establishment of abaxial-adaxial polarity in lateral organs involves factors intrinsic to the primordia and interactions with the apical meristem from which they are derived. Recently, a small plant-specific family of genes, the YABBY gene family, has been proposed to specify abaxial cell fate. Each asymmetric above-ground lateral organ expresses at least one member of the family in a polar manner, and loss- and gain-of-function studies indicate that they are sufficient to specify abaxial cell fate and that they act in both distinct and redundant manners. PMID- 10679448 TI - Three ways to learn the ABCs. AB - The ABC model of flower development represents a milestone in explaining how the fate of emerging floral organ primordia is specified. This model states that organ identity is specified by different combinations of the activities of the A, B and C class homeotic genes. In spite of the remarkable simplicity of this model, the complex regulatory interactions that establish the initial pattern of A, B and C gene activity have yet to be fully explained. It has been shown that the LEAFY gene functions early to promote flower meristem identity, and that it is subsequently required for the normal expression of the ABC genes. Recently, LEAFY has been identified as an immediate upstream regulator of the floral homeotic genes, thus opening up an avenue to examine the transcriptional interactions that underlie floral patterning. PMID- 10679449 TI - Genetics of gametophyte biogenesis in Arabidopsis. AB - The identification of several mutations and genes involved in sporogenesis and gametogenesis has initiated a genetic framework for understanding gametophyte biogenesis. Recent advances include the molecular characterization of genes required for sporocyte formation and meiosis. These studies have revealed some unexpected interactions linking development of sporophytic cells and tissues with initiation and progression of gametophyte development in angiosperms. PMID- 10679450 TI - Brassinosteroid signal transduction: still casting the actors. AB - Significant advances in the genetic dissection of brassinosteroid biosynthesis and signaling have been made during the past few years. Genetic and biochemical data have helped to elucidate the pathways of biosynthesis of brassinolide, the most active brassinosteroid. In addition, several models have been put forward for the perception of brassinolide by its putative receptor, BRI1, a ubiquitously expressed plasma membrane localized protein kinase. These studies provide the basic framework for future analysis of brassinosteroid signaling. PMID- 10679451 TI - New genes and new biological roles for expansins. AB - Expansins are extracellular proteins that loosen plant cell walls in novel ways. They are thought to function in cell enlargement, pollen tube invasion of the stigma (in grasses), wall disassembly during fruit ripening, abscission and other cell separation events. Expansins are encoded by two multigene families and each gene is often expressed in highly specific locations and cell types. Structural analysis indicates that one expansin region resembles the catalytic domain of family-45 endoglucanases but glucanase activity has not been detected. The genome projects have revealed numerous expansin-related sequences but their putative wall-loosening functions remain to be assessed. PMID- 10679452 TI - Developmental complexities of simple leaves. AB - Recent papers have explored early events in the development of simple leaves. Functional compartmentalization of the shoot apical meristem correlates with distinct fields of cells connected by plasmodesmata. Molecules important in the initiation of phyllotactic pattern are described and the relationship between dorsoventral patterning and lateral leaf expansion is investigated. PMID- 10679453 TI - Response of plant development to environment: control of flowering by daylength and temperature. AB - The transition from vegetative growth to flowering is often controlled by environmental conditions and influenced by the age of the plant. Intensive genetic analysis has identified pathways that regulate flowering time of Arabidopsis in response to daylength or low temperature (vernalization). These pathways are proposed to converge to regulate the expression of genes that act within the floral primordium and promote floral development. In the past year, genes that confer the responses to daylength or vernalization have been cloned and have enabled aspects of the genetic models to be tested at the molecular level. PMID- 10679454 TI - Division decisions and the spatial regulation of cytokinesis. AB - Cytokinesis in plant cells in accomplished when a membranous cell plate is guided to a pre-established division site. The orientation of the new wall establishes the starting position of a cell in a growing tissue, but the impact of this position on future development varies. Recently, proteins have been identified that participate in forming, stabilizing and guiding the cell plate to the correct division site. Mutations that affect cytokinesis with varying impacts on plant development are providing information about the mechanics of cytokinesis and also about how the division site is selected. PMID- 10679455 TI - Polarity: the role of localized secretion. AB - Studies of single cells from brown algae suggest that localized secretions stabilize the polar axis resulting in an asymmetry in the cell wall. This cortical asymmetry appears to play a role in orienting the plane of cell division and in determining the different fates of the resulting daughter cells. Recent studies indicate that similar processes may operate in seed plants. PMID- 10679456 TI - Enzymes that catalyse the restructuring of proteins. AB - The large enzyme families of protein disulfide isomerases and peptidyl prolyl cis/trans isomerases have been shown to assist polypeptide restructuring. Various folding states of polypeptides may serve as substrates of the catalysed reaction. Our understanding of the cellular function of these enzymes is increasing as a result of the availability of more specific inhibitors, the discovery of natural substrates and the use of genetically modified organisms. Further highlights of these studies include insights into the three-dimensional structures of enzyme ligand complexes, as well as into the mechanism of slow folding phases on the atomic level. PMID- 10679457 TI - DNA helicases: 'inching forward'. AB - Recently determined crystal structures of PcrA helicase complexed with a DNA substrate have revealed details of the helicase mechanism. PcrA and UvrD helicases have been shown to be functional as monomers, challenging previous suggestions that all helicases are required to be oligomeric. Crystal structures of the hexameric helicases RepA and T7 gene 4 explain the formation of hexameric assemblies from identical monomers with RecA-like folds, but their molecular mechanism remains elusive. PMID- 10679458 TI - The many routes of bacterial transfer RNAs after aminoacylation. AB - Subsequent to their aminoacylation, tRNAs are subject to specific maturation and/or correction processes. Aminoacylated tRNAs ready for use in translation are then specifically channelled to the ribosomal A or P sites. Structural and biochemical studies have opened the way towards furthering our understanding of these routes to the ribosome, which involve a strict distinction between initiator and elongator tRNAs. PMID- 10679459 TI - Structure and in vivo function of Hsp90. AB - Until recently, Hsp90 was one of the least well understood of the molecular chaperones, but considerable progress is now being made in unravelling its biochemistry. Hsp90 has now been shown to possess an inherent ATPase that is essential for the activation of authentic 'client' proteins in vivo and in vitro. The molecular detail of Hsp90's interactions with co-chaperones is also becoming clearer and the identification of key roles in assembling regulatory and signalling pathways has made it a target for anticancer drug development. Despite this, a clear understanding of how Hsp90 contributes to the folding and/or activation of its client proteins remains some way off. PMID- 10679460 TI - Mammalian prion proteins. AB - The past two years have seen the extension of our knowledge on the cellular prion protein structure with new NMR data on both the hamster and human proteins. In addition, the folding dynamics of two cellular prion proteins have been elucidated. There are now several examples of recombinant prion proteins that are able to adopt different conformations in solution and recent work on the molecular basis of prion strains has done much to consolidate the protein-only hypothesis. Important advances in relating disease to structure have also been made through the identification of the minimal prion protein fragment that is capable of conferring susceptibility to and propagation of the scrapie agent. PMID- 10679461 TI - Structural basis of mRNA cap recognition by proteins. AB - Crystal structures have recently become available for two proteins (VP39 and eIF4E) complexed with their cognate ligand - the mRNA cap. Despite their total structural dissimilarity, both proteins bind N7-methylguanine between two parallel aromatic sidechains. The resulting stacked arrangement governs their high specificity for the alkylated form of the nucleobase. PMID- 10679462 TI - Amyloid fibrillogenesis: themes and variations. AB - Recent progress has improved our knowledge of how proteins form amyloid fibrils. Both 'natively unfolded' and globular proteins have been shown to initiate fibrillization by adopting a partially structured conformation. Oligomeric prefibrillar intermediates have been extensively characterized with respect to their morphology and temporal evolution. Three-dimensional models obtained using biophysical and computational methods have provided information about fibril structure. All of these advances suggest common features of self-assembly pathways, with subtle variations accounting for differences among distinct amyloid fibrils. PMID- 10679463 TI - Protein folding mechanisms: new methods and emerging ideas. AB - During the past year, advances in our understanding of folding mechanisms have been made through detailed experimental and theoretical studies of a number of proteins. The development of new methods has allowed the earliest events in folding to be probed and the measurement of folding at the level of individual molecules is now possible, opening the door to exciting new experiments. PMID- 10679464 TI - Small heat-shock proteins and their potential role in human disease. AB - The elevated expression of stress proteins is considered to be a universal response to adverse conditions, representing a potential mechanism of cellular defense against disease and a potential target for novel therapeutics, including gene therapy and chaperone-modulating reagents. Recently, a single mutation in the small heat-shock protein human alphaB-crystallin was linked to desmin-related myopathy, which is characterized by abnormal intracellular aggregates of intermediate filaments in human muscle. New findings demonstrate that the high level of expression of stress proteins can contribute to an autoimmune response and can protect proteins that contribute to disease processes. PMID- 10679465 TI - Implications of macromolecular crowding for protein assembly. AB - Recent studies have led to increased appreciation of the influence of excluded volume in solutions of high total macromolecular content ('macromolecular crowding') upon the various classes of reaction that lead to the assembly of proteins and protein complexes. In general, crowding is expected to stabilize native protein structure relative to less compact non-native structures and to favor the formation of functional complexes of native proteins. Under certain pathological conditions, 'overcrowding' may enhance the formation of nonfunctional aggregates of non-native protein (e.g. amyloid and inclusion bodies). PMID- 10679466 TI - Single-stranded-RNA binding proteins. AB - Our knowledge of protein interactions with RNA molecules has been, so far, largely restricted to cases in which the RNA itself is folded into a secondary and/or tertiary structure stabilised by intramolecular base pairing and stacking. Until recently, only limited structural information has been available about protein interactions with single-stranded RNA. A breakthrough in our understanding of these interactions came in 1999, with the determination of four crystal structures of protein complexes with extended single-stranded RNA molecules. These structures revealed wonderfully satisfying patterns of the ability of proteins to accommodate RNA bases, with the sugar-phosphate backbone often adopting conformations that are different from the classical double helix. PMID- 10679467 TI - Protein folding in vivo: the importance of molecular chaperones. AB - The contribution of the two major cytosolic chaperone systems, Hsp70 and the cylindrical chaperonins, to cellular protein folding has been clarified by a number of recent papers. These studies found that, in vivo, a significant fraction of newly synthesized polypeptides transit through these chaperone systems in both prokaryotic and eukaryotic cells. The identification and characterization of the cellular substrates of chaperones will be instrumental in understanding how proteins fold in vivo. PMID- 10679468 TI - Insights into transcription: structure and function of single-subunit DNA dependent RNA polymerases. AB - Single-subunit RNA polymerases are widespread throughout prokaryotic and eukaryotic organisms, and also viruses. T7 RNA polymerase is one of the simplest DNA-dependent enzymes, capable of transcribing a complete gene without the need for additional proteins. During the past two years, three illuminating crystal structures of T7 RNA polymerase complexed to either T7 lysozyme, which is a transcription inhibitor, an open promoter DNA fragment or a promoter DNA fragment being transcribed into RNA at initiation have been determined. For the first time, these structures describe in detail the intricate mechanism of transcription initiation by T7 RNA polymerase, which is likely to be a general model for other related RNA polymerases. PMID- 10679469 TI - Recognition of distorted DNA structures by HMG domains. AB - Recent biochemical and structural studies have shown that the preferential recognition of distorted DNA structures, including DNA bulges, four-way junctions and cis-platinated DNA, by HMG domains is dependent on residues immediately preceding the second alpha helix of the L-shaped HMG domain. PMID- 10679470 TI - Winged helix proteins. AB - The winged helix proteins constitute a subfamily within the large ensemble of helix-turn-helix proteins. Since the discovery of the winged helix/fork head motif in 1993, a large number of topologically related proteins with diverse biological functions have been characterized by X-ray crystallography and solution NMR spectroscopy. Recently, a winged helix transcription factor (RFX1) was shown to bind DNA using unprecedented interactions between one of its eponymous wings and the major groove. This surprising observation suggests that the winged helix proteins can be subdivided into at least two classes with radically different modes of DNA recognition. PMID- 10679471 TI - Mysteries of magnesium homeostasis. PMID- 10679472 TI - Myoendothelial gap junctions: the gap is there, but does EDHF go through it? PMID- 10679473 TI - Vascular endothelial growth factor: A Jack-of-all-trades or a nonspecific stress gene? PMID- 10679474 TI - Viral myocarditis: receptors that bridge the cardiovascular with the immune system? PMID- 10679475 TI - Calcineurin-mediated hypertrophy protects cardiomyocytes from apoptosis in vitro and in vivo: An apoptosis-independent model of dilated heart failure. AB - We have previously shown that the calcium-calmodulin-regulated phosphatase calcineurin (PP2B) is sufficient to induce cardiac hypertrophy that transitions to heart failure in transgenic mice. Given the rapid onset of heart failure in these mice, we hypothesized that calcineurin signaling would stimulate myocardial cell apoptosis. However, utilizing multiple approaches, we determined that calcineurin-mediated hypertrophy protected cardiac myocytes from apoptosis, suggesting a model of heart failure that is independent of apoptosis. Adenovirally mediated gene transfer of a constitutively active calcineurin cDNA (AdCnA) was performed in cultured neonatal rat cardiomyocytes to elucidate the mechanism whereby calcineurin affected myocardial cell viability. AdCnA infection, which induced myocyte hypertrophy and atrial natriuretic factor expression, protected against apoptosis induced by 2-deoxyglucose or staurosporine, as assessed by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) labeling, caspase-3 activation, DNA laddering, and cellular morphology. The level of protection conferred by AdCnA was similar to that of adenoviral Bcl-x(L) gene transfer or hypertrophy induced by phenylephrine. In vivo, failing hearts from calcineurin-transgenic mice did not demonstrate increased TUNEL labeling and, in fact, demonstrated a resistance to ischemia/reperfusion-induced apoptosis. We determined that the mechanism whereby calcineurin afforded protection from apoptosis was partially mediated by nuclear factor of activated T cells (NFAT3) signaling and partially by Akt/protein kinase B (PKB) signaling. Although calcineurin activation protected myocytes from apoptosis, inhibition of calcineurin with cyclosporine was not sufficient to induce TUNEL labeling in Gqalpha-transgenic mice or in cultured cardiomyocytes. Collectively, these data identify a calcineurin-dependent mouse model of dilated heart failure that is independent of apoptosis. PMID- 10679476 TI - Targeted disruption of the mouse Sod I gene makes the hearts vulnerable to ischemic reperfusion injury. AB - The role of Cu/Zn-superoxide dismutase (SOD) in myocardial ischemic reperfusion injury was studied by using a mouse model with targeted disruption of the mouse Sod I gene. Inactivation of the functional mouse Sod I gene in hearts by gene targeting (Sod I(+/-)) resulted in a 50% reduction of Cu/Zn-SOD mRNA and significant reduction of Cu/Zn-SOD enzyme activity compared with that of wild type Sod I(+/+) mice. Cu/Zn-SOD mRNA could not be detected in Sod I(-/-) heart. The isolated buffer-perfused hearts from the knockout mice devoid of any functional copy of the Sod I (Sod I(-/-)) and matched nontransgenic control mice were subjected to 30 minutes of global ischemia followed by 2 hours of reperfusion. For both groups of mice, the postischemic functional recovery for the hearts was lower than the baseline, but the recovery for the Sod I(-/-) was less compared with the wild-type mice. Thus, the postischemic recovery of the developed force and the maximum first derivative of the developed force were consistently lower for the Sod I(-/-) mouse hearts compared with wild-type control hearts. The coronary flow was lower compared with the baseline levels for both groups of hearts, but there was no significant difference between the groups. The myocardial infarction determined from the ratio of infarct size/area of risk was higher for the Sod I(-/-) mice compared with the control mice. The amount of creatine kinase release from the wild-type mouse hearts was less compared with the Sod I(-/-) mouse hearts. In concert, a reduced amount of oxidative stress was found in the hearts of wild-type mice compared with Sod I(-/ ) mouse hearts. These results documented that Sod I(-/-) mouse hearts were more susceptible to ischemic reperfusion injury compared with corresponding wild-type mouse hearts, suggesting that the Sod I gene constitutes an important defense element for the hearts. PMID- 10679477 TI - Myocardial glucose uptake is regulated by nitric oxide via endothelial nitric oxide synthase in Langendorff mouse heart. AB - Although the role of nitric oxide (NO) in the modulation of vascular tone has been studied and well understood, its potential role in the control of myocardial metabolism is only recently evident. Several lines of evidence indicate that NO regulates myocardial glucose metabolism; however, the details and mechanisms responsible are still unknown. The aim of this study was to further define the role of NO in the control of myocardial glucose metabolism and the nitric oxide synthase (NOS) isoform responsible using transgenic animals lacking endothelial NOS (ecNOS). In the present study, we examined the regulation of myocardial glucose uptake using isometrically contracting Langendorff-perfused hearts from normal mice (C57BL/6J), mice with defects in the expression of ecNOS [ecNOS (-/ )], and its heterozygote [ecNOS (+/-)], and wild-type mice [ecNOS (+/+)] (n=6, respectively). In hearts from normal mice, little myocardial glucose uptake was observed. This myocardial glucose uptake increased significantly in the presence of N(omega)-nitro-L-arginine methyl ester (L-NAME). Similarly, in the hearts from ecNOS (-/-), glucose uptake was much greater than in normal mice, whereas myocardial glucose uptake of ecNOS (+/-) and ecNOS (+/+) mice was not different from normal mice. In addition, myocardial glucose uptake of ecNOS (+/-) and ecNOS (+/+) mice increased significantly in the presence of L-NAME. At a workload of 800 g. beats/min, L-NAME increased glucose uptake from 0.1+/-0.1 to 3+/-0.4 microg/min x mg in ecNOS (+/-) mice and from 0.2+/-0.1 to 2.7+/-0.7 microg/min x mg in ecNOS (+/+) mice. Furthermore, in the hearts from ecNOS (-/-) mice, 8 bromoguanosine 3':5'-cyclic monophosphate (8-Br-cGMP), a cGMP analog or S-nitroso N-acetylpenicillamine (SNAP), a NO donor essentially shut off glucose uptake, and in hearts from ecNOS (+/-) mice, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ), an inhibitor of cGMP, increased the glucose uptake significantly. These results indicate clearly that cardiac NO production regulates myocardial glucose uptake via a cGMP-dependent mechanism and strongly suggest that ecNOS plays a pivotal role in this regulation. These findings may be important in the understanding of the pathogenesis of the diseases such as ischemic heart disease, heart failure, diabetes mellitus, hypertension, and hypercholesterolemia, in which NO synthesis is altered and substrate utilization by the heart changes. PMID- 10679478 TI - Expression of coxsackievirus and adenovirus receptor in hearts of rats with experimental autoimmune myocarditis. AB - The expression of coxsackievirus and adenovirus receptor (CAR) was dominant in the brains and hearts of mice until the newborn phase. There is no detailed information concerning the relation between the expression of CAR and development of hearts. It is also uncertain whether CAR is able to be induced in adult hearts after cardiac injury. We demonstrated that CAR was abundant in the hearts of newborn rats but was barely detectable in the hearts of adult rats. The expression of CAR in rat hearts with experimental autoimmune myocarditis, which was induced by immunization of purified cardiac myosin, was serially investigated. Active myocarditis was observed from day 15 after immunization. By immunohistochemistry, cardiomyocytes were strongly stained for CAR antibody from days 24 to 42. CAR mRNA was also detected from days 18 to 30 by using reverse transcription-polymerase chain reaction. In the next experiment, the induction of CAR on isolated cardiomyocytes was investigated. CAR was barely detectable in cultured cardiomyocytes by Western blot analysis after isolation. This molecule gradually appeared along with the creation of clusters and beating of cardiomyocytes. Furthermore, the induction of CAR in cultured cardiomyocytes increased after supplement with conditioned medium of rat splenocytes activated by concanavalin A. In conclusion, rat CAR is expressed strongly in the hearts of newborn rats and is suppressed in those of adult rats. The expression of CAR is enhanced during the active phase of experimental autoimmune myocarditis and is induced by inflammatory mediators. CAR may play a role in cell-to-cell contact and adhesion of cardiomyocytes. PMID- 10679479 TI - Immunoglobulin isotype determines pathogenicity in antibody-mediated myocarditis in naive mice. AB - Antimyosin reactivity is associated with cardiac damage in autoimmune myocarditis, an inflammatory heart disease characterized by a cellular infiltrate in the myocardium and myocyte necrosis. We are interested in the pathogenicity of antimyosin antibodies and their ability to cause autoimmune myocarditis. We have shown that antimyosin antibodies of the IgG isotype will induce disease in the DBA/2 mouse. In the present study, we show that IgM antimyosin antibodies do not induce myocarditis; however, these same antibodies become pathogenic when converted to the IgG isotype. Although IgM antibodies can penetrate the myocardium during cardiac inflammation, they are usually less able to leave the vascular compartment and penetrate cardiac tissue, thus accounting for their lack of pathogenicity. Thus, antimyosin B cells may be potentially pathogenic only after antigen activation and heavy chain class switching or under conditions that alter vascular permeability in the heart. PMID- 10679480 TI - Intussusceptive angiogenesis: its role in embryonic vascular network formation. AB - Intussusceptive angiogenesis is a novel mode of blood vessel formation and remodeling, which occurs by internal division of the preexisting capillary plexus without sprouting. In this study, the process is demonstrated in developing chicken eye vasculature and in the chorioallantoic membrane by methylmethacrylate (Mercox) casting, transmission electron microscopy, and in vivo observation. In a first step of intussusceptive angiogenesis, the capillary plexus expands by insertion of numerous transcapillary tissue pillars, ie, by intussusceptive microvascular growth. In a subsequent step, a vascular tree arises from the primitive capillary plexus as a result of intussusceptive pillar formation and pillar fusions, a process we termed "intussusceptive arborization." On the basis of the morphological observations, a 4-step model for intussusceptive arborization is proposed, as follows: phase I, numerous circular pillars are formed in rows, thus demarcating future vessels; phase II, formation of narrow tissue septa by pillar reshaping and pillar fusions; phase III, delineation, segregation, growth, and extraction of the new vascular entity by merging of septa; and phase IV, formation of new branching generations by successively repeating the process, complemented by growth and maturation of all components. In contrast to sprouting, intussusceptive angiogenesis does not require intense local endothelial cell proliferation; it is implemented primarily by rearrangement and attenuation of the endothelial cell plates. In summary, transcapillary pillar formation, ie, intussusception, is a central and probably widespread process, which plays a role not only in capillary network growth and expansion (intussusceptive microvascular growth), but also in vascular plexus remodeling and tree formation (intussusceptive arborization). PMID- 10679481 TI - The epsilon subtype of protein kinase C is required for cardiomyocyte connexin-43 phosphorylation. AB - Gap junctions (GJs), composed of connexins, are intercellular channels ensuring electric and metabolic coupling between cardiomyocytes. We have shown previously that an endogenous mitogenic and cardioprotective protein, fibroblast growth factor-2 (FGF-2), decreases cardiomyocyte GJ permeability by stimulating phosphorylation of connexin-43 (Cx43). Identifying the kinase(s) phosphorylating cardiac Cx43 may thus provide a way of modulating cardiac intercellular communication. Because FGF-2 activates receptors linked to protein kinase C (PKC) and mitogen-activated protein kinase, we first investigated participation of these enzymatic systems in Cx43 phosphorylation. The inhibitor PD98059 blocked activation of mitogen-activated protein kinase, but it did not prevent the FGF-2 effects on GJs. In contrast, the PKC inhibitor chelerythrine blocked the effects of FGF-2 on Cx43 phosphorylation and permeability. Because the epsilon-isoform of PKC localizes to plasma membrane sites, we examined whether it is directly involved in the FGF-2-induced Cx43 phosphorylation. In nonstimulated myocytes, PKCepsilon displayed a discontinuous pattern of localization at intercellular contact sites and partial colocalization with Cx43. Treatment with FGF-2 or phorbol 12-myristate 13-acetate induced a more continuous pattern of PKCepsilon distribution, whereas the anti-Cx43 staining appeared to overlap extensively with that of PKCepsilon. In immunoprecipitation experiments using specific anti-Cx43 antibodies, PKCepsilon but not PKCalpha coprecipitated with Cx43. FGF-2 increased levels of coprecipitated PKCepsilon, suggesting increased association between PKCepsilon and Cx43 on stimulation. Transient gene transfer and overexpression of cDNAs coding for truncated or mutated dominant-negative forms of PKCepsilon decreased cardiomyocyte Cx43 phosphorylation significantly. We conclude that PKC mediates the FGF-2-induced effects on cardiac GJs and that PKCepsilon likely interacts with and phosphorylates cardiac Cx43 at sites of intercellular contact. PMID- 10679482 TI - Electrophysiological effects of remodeling cardiac gap junctions and cell size: experimental and model studies of normal cardiac growth. AB - The increased incidence of arrhythmias in structural heart disease is accompanied by remodeling of the cellular distribution of gap junctions to a diffuse pattern like that of neonatal cardiomyocytes. Accordingly, it has become important to know how remodeling of gap junctions due to normal growth hypertrophy alters anisotropic propagation at a cellular level (V(max)) in relation to conduction velocities measured at a macroscopic level. To this end, morphological studies of gap junctions (connexin43) and in vitro electrical measurements were performed in neonatal and adult canine ventricular muscle. When cells enlarged, gap junctions shifted from the sides to the ends of ventricular myocytes. Electrically, normal growth produced different patterns of change at a macroscopic and microscopic level. Although the longitudinal and transverse conduction velocities were greater in adult than neonatal muscle, the anisotropic velocity ratios were the same. In the neonate, mean V(max) was not different during longitudinal (LP) and transverse (TP) propagation. However, growth hypertrophy produced a selective increase in mean TP V(max) (P<0.001), with no significant change in mean LP V(max). Two-dimensional neonatal and adult cellular computational models show that the observed increases in cell size and changes in the distribution of gap junctions are sufficient to account for the experimental results. Unexpectedly, the results show that cellular scaling (cell size) is as important (or more so) as changes in gap junction distribution in determining TP properties. As the cells enlarged, both mean TP V(max) and lateral cell-to-cell delay increased. V(max) increased because increases in cell-to-cell delay reduced the electric current flowing downstream up to the time of V(max), thus enhancing V(max). The results suggest that in pathological substrates that are arrhythmogenic, maintaining cell size during remodeling of gap junctions is important in sustaining a maximum rate of depolarization. PMID- 10679483 TI - Acidosis inhibits endothelial cell apoptosis and function and induces basic fibroblast growth factor and vascular endothelial growth factor expression. AB - Endothelial cells are exposed to an acidotic environment in a variety of pathological and physiological conditions. However, the effect of acidosis on endothelial cell function is still largely unknown, and it was evaluated in the present study. Bovine aortic endothelial cells (BAECs) were grown in bicarbonate buffer equilibrated either with 20% CO(2) (pH 7.0, acidosis) or 5% CO(2) (pH 7.4, control). Acidosis inhibited BAEC proliferation in 10% FCS, whereas by day 7 in serum-free medium, cell number was 3-fold higher in acidotic cells than in control cells. Serum deprivation enhanced BAEC apoptosis, and apoptotic cell death was markedly inhibited by acidosis. Additionally, acidosis inhibited FCS stimulated migration in a modified Boyden chamber assay and FCS-stimulated differentiation into capillary-like structures on reconstituted basement membrane proteins. Conditioned media from BAECs cultured for 48 hours either at pH 7.0 or pH 7.4 enhanced BAEC proliferation and migration at pH 7.4, and both effects were more marked with conditioned medium from BAECs grown in acidotic than in control conditions. Acidosis enhanced vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA expression as well as bFGF secretion, and a blocking bFGF antibody inhibited enhanced BAEC migration in response to conditioned medium from acidotic cells. These results show that acidosis protects endothelial cells from apoptosis and inhibits their proangiogenic behavior despite enhanced VEGF and bFGF mRNA expression and bFGF secretion. PMID- 10679484 TI - Hypoxic regulation of inducible nitric oxide synthase via hypoxia inducible factor-1 in cardiac myocytes. AB - The relationship between hypoxia and regulation of nitric oxide synthase (NOS) in myocardial tissue is not well understood. We investigated the role of hypoxia inducible factor-1 (HIF-1) on expression of the inducible NOS (iNOS) in myocardial cells in vivo and in vitro. In situ hybridization in myocardial tissue from rats exposed to hypoxia for 3 weeks demonstrated increased iNOS mRNA expression. Northern analysis of RNA from hearts of those animals and from cells exposed to hypoxia for 12 hours in vitro demonstrated an increase of HIF-1 RNA expression. Electrophoretic mobility shift assays using oligonucleotides containing the iNOS HIF-1 DNA binding site and nuclear extracts from cardiac myocytes showed induction of specific DNA binding in cells subjected to hypoxia. Transient transfection of cardiac myocytes using the murine iNOS promoter resulted in a 3.43-fold increase in promoter activity under hypoxia compared with normoxia. Mutation or deletion of the HIF-1 site eliminated the hypoxic response. As cytokines have been shown to regulate iNOS expression in myocardial cells, cultured neonatal cardiac myocytes were stimulated with interleukin-1beta causing a dramatic induction of iNOS protein expression under normoxia, with further augmentation under hypoxia. Transient transfection of cells stimulated with interleukin-1beta showed an increased iNOS promoter activity under normoxic conditions compared with unstimulated cells, with a further increase in response to hypoxia, which was dependent on HIF-1. These results demonstrate that hypoxia causes an increase in iNOS expression in cardiac myocytes and that HIF-1 is essential for the hypoxic regulation of iNOS gene expression. PMID- 10679485 TI - Parallel stimulation of glucose and Mg(2+) accumulation by insulin in rat hearts and cardiac ventricular myocytes. AB - The stimulation of beta-adrenoceptors in cardiac cells results in a rapid loss of cellular Mg(2+). Because insulin physiologically counteracts several of the cellular effects mediated by the activation of beta-adrenoceptors and the elevation of cytosolic cAMP levels, we investigated whether insulin administration could prevent Mg(2+) mobilization from rat hearts and ventricular myocytes. Rat hearts were perfused in a retrograde Langendorff system, and the changes in extracellular Mg(2+) were measured by atomic absorbance spectrophotometry. Pretreatment of the hearts with 6 nmol/L insulin completely prevented the Mg(2+) extrusion induced by the beta-adrenergic agonist isoproterenol. Furthermore, the administration of insulin per se induced an accumulation of Mg(2+) by the heart. This accumulation was small but detectable in the presence of 25 to 35 micromol/L [Mg(2+)](o) and increased in proportion to [Mg(2+)](o). Insulin-mediated Mg(2+) accumulation was not observed in hearts perfused with a medium devoid of glucose or with a medium containing the inhibitors of glucose transport, cytochalasin B and phloretin. Insulin-stimulated [(3)H]2-deoxyglucose accumulation was measured in collagenase-dispersed cardiac ventricular myocytes in the presence of varying levels of [Mg(2+)](o). Glucose transport was not observed below 25 micromol/L [Mg(2+)](o), and it also increased in proportion to [Mg(2+)](o). Taken together, these results indicate the presence of a major uptake of Mg(2+) into cardiac cells that is stimulated by insulin and may require the insulin-induced operation of a glucose transporter. Hence, extracellular and/or intracellular Mg(2+) may modulate glucose transport and/or utilization. PMID- 10679486 TI - Photodynamic therapy generates a matrix barrier to invasive vascular cell migration. AB - Photodynamic therapy (PDT) inhibits experimental intimal hyperplasia. PDT results in complete vascular wall cell eradication with subsequent adventitia but minimal media repopulation. This study was designed to test the hypothesis that PDT alters the vascular wall matrix thereby inhibiting invasive cell migration, and as such, provides an important barrier mechanism to favorably alter the vascular injury response. Untreated smooth muscle cells (SMCs) and fibroblasts were seeded on control and PDT-treated (100 J/cm(2); photosensitizer was chloroaluminum sulfonated phthalocyanine, 5 microg/mL) 3-dimensional collagen matrix gels. Invasive cell migration was temporally quantified by calibrated microscopy. Zymography and ELISA assessed SMC matrix metalloproteinase levels. Molecular changes of gel proteins and their susceptibility to collagenase were analyzed by SDS-PAGE and Western blot. Limited pepsin digestion and histology were used to assess the in vivo relevance of the model, using an established rat carotid artery model at 1 and 4 weeks after balloon injury and PDT. PDT of 3-dimensional matrix of gels led to a 52% reduction of invasive SMCs and to a 59% reduction of fibroblast migration (P<0.001) but did not significantly affect secretion of matrix metalloproteinases. PDT induced collagen matrix changes, including cross linking, which resulted in resistance to protease digestion. PDT led to a durable 45% reduction in pepsin digestion susceptibility of treated arteries (P<0.001) and inhibition of periadventitial cell migration into the media. These data suggest that PDT of matrix gels generates a barrier to invasive cellular migration. This newly identified effect on matrix proteins underscores its pleiotropic actions on the vessel wall, and as such, PDT may be of considerable potential therapeutic value to inhibit restenosis. PMID- 10679487 TI - Incidence of myoendothelial gap junctions in the proximal and distal mesenteric arteries of the rat is suggestive of a role in endothelium-derived hyperpolarizing factor-mediated responses. AB - Although the chemical nature of endothelium-derived hyperpolarizing factor (EDHF) remains elusive, electrophysiological evidence exists for electrical communication between smooth muscle cells and endothelial cells suggesting that electrotonic propagation of hyperpolarization may explain the failure to identify a single chemical factor as EDHF. Anatomical evidence for myoendothelial gap junctions, or the sites of electrical coupling, is, however, rare. In the present study, serial-section electron microscopy and reconstruction techniques have been used to examine the incidence of myoendothelial gap junctions in the proximal and distal mesenteric arteries of the rat where EDHF responses have been reported to vary. Myoendothelial gap junctions were found to be very small in the mesenteric arteries, the majority being <100 nm in diameter. In addition, they were significantly more common in the distal compared with the proximal regions of this arterial bed. Pentalaminar gap junctions between adjacent endothelial cells were much larger and were common in both proximal and distal mesenteric arteries. These latter junctions were frequently found near the myoendothelial gap junctions. These results provide the first evidence for the presence of sites for electrical communication between endothelial cells and smooth muscle cells in the mesenteric vascular bed. Furthermore, the relative incidence of these sites suggests that there may be a relationship between the activity of EDHF and the presence of myoendothelial gap junctions. PMID- 10679488 TI - Transcriptional and posttranscriptional regulation of endothelial nitric oxide synthase expression by hydrogen peroxide. AB - Diverse stimuli, including shear stress, cyclic strain, oxidized LDL, hyperglycemia, and cell growth, modulate endothelial nitric oxide synthase (eNOS) expression. Although seemingly unrelated, these may all alter cellular redox state, suggesting that reactive oxygen intermediates might modulate eNOS expression. The present study was designed to test this hypothesis. Exposure of bovine aortic endothelial cells for 24 hours to paraquat, a superoxide (O(2)(-*)) generating compound, did not affect eNOS mRNA levels. However, cotreatment with paraquat and either Cu(2+)/Zn(2+) superoxide dismutase or the superoxide dismutase mimetic tetrakis(4-benzoic acid)porphyrin chloride increased eNOS mRNA by 2.3- and 2.2-fold, respectively, implicating a role for H(2)O(2). Direct addition of 100 and 150 micromol/L H(2)O(2) caused increases in bovine aortic endothelial cell eNOS mRNA that were dependent on concentration (ie, 3.1- and 5.2 fold increases) and time, and elevated eNOS protein expression and enzyme activity, accordingly. Nuclear run-on and 5, 6-dichloro-1-beta-D ribofuranosylbenzimidazole-chase studies showed that H(2)O(2) caused a 3.0-fold increase in eNOS gene transcription and a 2.8-fold increase in eNOS mRNA half life. Induction of eNOS by H(2)O(2) was not affected by the hydroxyl radical scavenger DMSO, mannitol, or N-tert-butyl-alpha-phenylnitrone, but it was inhibited by the antioxidants N-acetylcysteine, ebselen, and exogenously added catalase. Unlike H(2)O(2), the 4.0-fold induction of eNOS by shear stress (15 dyne/cm(2) for 6 hours) was not inhibited by N-acetylcysteine or exogenous catalase. In conclusion, H(2)O(2) increases eNOS expression through transcriptional and post-transcriptional mechanisms. Although H(2)O(2) does not mediate shear-dependent eNOS regulation, it is likely to be involved in regulation of eNOS expression in response to other physiological and/or pathophysiological stimuli. PMID- 10679489 TI - Interactions between Ca(2+) and H(+) and functional consequences in vascular smooth muscle. AB - Ca(2+) and H(+) ions can profoundly alter vascular tone. In many physiological and pathological processes, changes in the concentration of both ions occur. Thus, to understand the processes and mechanisms that modify force, it is necessary to understand what changes occur in these ions and, importantly, how they interact with each other. In this minireview, we highlight the quantitatively important mechanisms involved in the contractile responses of vascular tissues to pH change and discuss the cellular and molecular reasons underlying these responses. PMID- 10679490 TI - Regulation of endothelial nitric oxide synthase expression by albumin-derived advanced glycosylation end products. AB - We examined whether albumin-derived advanced glycosylation end products (AGEs) downregulate the expression of endothelial nitric oxide synthase (NOS). Significant reductions in NOS activity and cGMP levels in bovine aortic endothelial cells were observed when exposed to different concentrations of albumin-derived AGEs. Western and Northern blot analyses showed significant decreases at the protein and transcript levels. Both reductions became evident after 24 hours of exposure. Nuclear run-on assays showed that AGE-BSA did not modify the transcription rate of the NOS III gene; however, AGE-BSA treatment markedly reduced the half-life of NOS III mRNA. In addition, AGE-treated endothelial cells displayed significant reduction on their antiplatelet properties. These results indicate that NOS expression is reduced by AGEs by increasing the rate of mRNA degradation and may be relevant to the impairment of some endothelial functions observed in diabetes and aging. The full text of this article is available at http://www.circresaha.org. PMID- 10679491 TI - Possible mechanism(s) of arachidonic acid-induced intracellular acidosis in rat cardiac myocytes. AB - Arachidonic acid (AA) and other nonesterified fatty acids (FAs) have been shown to exert harmful effects during cardiac ischemia. By continuously measuring intracellular pH (pH(i)) changes in neonatal and adult cardiac myocytes, we have found, for the first time, that 10 micromol/L AA induces a substantial intracellular acidosis (0.3 to 0.4 pH units). We have ruled out the possibilities that the AA-induced acidosis is caused by (1) inhibition or stimulation of the pH(i) regulators, (2) protein kinase C activation or the generation of AA metabolites or free radicals, or (3) activation of NADPH oxidase or an inward H(+) current. The AA-induced acidosis fits to a simple diffusion mechanism, as proposed by Kamp and Hamilton (flip-flop model) for artificial phospholipid bilayers. The important properties found in the cardiac myocyte are that (1) the initial rate of acid flux (J(H)) increases with the AA concentration (2 to 50 micromol/L), (2) FAs with a (-)COOH group (eg, AA, oleic acid, and linoleic acid) induce intracellular acidification, but FAs with a (-)COOCH(3) group (eg, AA methyl ester) have little effect on the pH(i), (3) tetradecylamine (FA amine) induces intracellular alkalosis, and, most importantly, (4) both the AA- and tetradecylamine-induced pH(i) changes can be reversed by 0.3% BSA. Because a low concentration of AA (10 micromol/L) can induce a substantial acidosis, the possible involvement of the FA-evoked acidosis in the negative inotropic effect during cardiac ischemia is discussed. The full text of this article is available at http://www. circresaha.org. PMID- 10679492 TI - Online-only or Print? : An urgent request to readers and authors PMID- 10679493 TI - e-hypertension : opening new vistas PMID- 10679494 TI - Long-term absolute benefit of lowering blood pressure in hypertensive patients according to the JNC VI risk stratification. AB - Blood pressure (BP) levels alone have been traditionally used to make treatment decisions in patients with hypertension. The sixth report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) recently recommended that risk strata, in addition to BP levels, be considered in the treatment of hypertension. We estimated the absolute benefit associated with a 12 mm Hg reduction in systolic BP over 10 years according to the risk stratification system of JNC VI using data from the National Health and Nutrition Examination Survey Epidemiologic Follow-up Study. The number-needed-to-treat to prevent a cardiovascular event/death or a death from all causes was reduced with increasing levels of baseline BP in each of the risk strata. In addition, the number-needed-to-treat was much smaller in persons with > or =1 additional major risk factor for cardiovascular disease (risk group B) and in those with a history of cardiovascular disease or target organ damage (risk group C) than in those without additional major risk factors for cardiovascular disease (risk group A). Specifically, the number-needed-to-treat to prevent a death from all causes in patients with a high-normal BP, stage 1 hypertension, or stage 2 or 3 hypertension was, respectively, 81, 60, and 23 for those in risk group A; 19, 16, and 9 for those in risk group B; and 14, 12, and 9 for those in risk group C. Our analysis indicated that the absolute benefits of antihypertensive therapy depended on BP as well as the presence or absence of additional cardiovascular disease risk factors and the presence or absence of preexisting clinical cardiovascular disease or target organ damage. PMID- 10679495 TI - Long-term effects of weight loss and dietary sodium reduction on incidence of hypertension. AB - To examine the long-term effects of weight loss and dietary sodium reduction on the incidence of hypertension, we studied 181 men and women who participated in the Trials of Hypertension Prevention, phase 1, in Baltimore, Md. At baseline (1987 to 1988), subjects were 30 to 54 years old and had a diastolic blood pressure (BP) of 80 to 89 mm Hg and systolic BP <160 mm Hg. They were randomly assigned to one of two 18-month lifestyle modification interventions aimed at either weight loss or dietary sodium reduction or to a usual care control group. At the posttrial follow-up (1994 to 1995), BP was measured by blinded observers who used a random-zero sphygmomanometer. Incident hypertension was defined as systolic BP > or =160 mm Hg and/or diastolic BP > or =90 mm Hg and/or treatment with antihypertensive medication during follow-up. Body weight and urinary sodium were not significantly different among the groups at the posttrial follow-up. After 7 years of follow-up, the incidence of hypertension was 18.9% in the weight loss group and 40.5% in its control group and 22.4% in the sodium reduction group and 32.9% in its control group. In logistic regression analysis adjusted for baseline age, gender, race, physical activity, alcohol consumption, education, body weight, systolic BP, and urinary sodium excretion, the odds of hypertension was reduced by 77% (odds ratio 0.23; 95% confidence interval 0.07 to 0.76; P=0.02) in the weight loss group and by 35% (odds ratio 0.65; 95% confidence interval 0.25 to 1.69; P=0.37) in the sodium reduction group compared with their control groups. These results indicate that lifestyle modification such as weight loss may be effective in long-term primary prevention of hypertension. PMID- 10679496 TI - Regulation of sodium balance and blood pressure by the AT(1A) receptor for angiotensin II. AB - To examine the role of the angiotensin II (AT)(1A) receptor in the regulation of blood pressure and sodium balance, we measured systolic blood pressure responses in AT(1A) receptor-deficient (Agtr1a-/-) and wild-type (Agtr1a+/+) mice while dietary sodium content was systematically altered. On a 0.4% sodium diet, systolic blood pressures were significantly lower in Agtr1a-/- than in +/+ mice. In Agtr1a+/+ mice, changing dietary sodium content did not affect blood pressure. In contrast, when Agtr1a-/- mice were fed a high-salt diet (6% NaCl), their systolic blood pressures increased significantly from 79+/-4 to 94+/-4 mm Hg (P<0.006). The low blood pressures of Agtr1a-/- mice decreased further while on a low-salt diet from 82+/-3 to 69+/-3 mm Hg (P<0.03). On the high-salt diet, urinary sodium excretion increased to similar levels in Agtr1a+/+ and -/- mice. Although urinary sodium excretion was substantially reduced in both groups during the low-salt diet, cumulative sodium balances became negative in Agtr1a-/- mice despite a 6-fold increase in urinary aldosterone. We infer, therefore, that the reduced blood pressures in Agtr1a-/- mice on a normal diet are caused by depletion of sodium and extracellular volume. Their "sodium sensitivity" suggests a critical role for renal AT(1A) receptors to modulate sodium handling. PMID- 10679497 TI - beta-2 adrenergic receptor gene variations, blood pressure, and heart size in normal twins. AB - Genetic variability, which influences cardiovascular phenotypes in normal persons, is likely to be relevant to cardiovascular disease. We studied normal monozygotic and dizygotic twins and found strong genetic influences on blood pressure and heart size. We then relied on the dizygotic twins and their parents to apply molecular genetic techniques. We performed a linkage analysis with markers close to the beta-2 adrenergic receptor (AR) gene locus in the dizygotic twins and their parents and found strong evidence for linkage to the quantitative traits of blood pressure and heart size. We then used allele-specific polymerase chain reaction to genotype the subjects further. We performed an association analysis and found that 4 functionally relevant polymorphisms in the beta-2 AR gene, namely Arg16/Gly, Gln27/Glu, Thr164/Ile, and a variant in the promoter region (-47C/T), were variably associated with blood pressure and heart size differences but were in linkage dysequilibrium with each other. A subsequent conditional analysis suggested that the Arg16/Gly polymorphism exerted the predominant effect. These findings underscore the importance of the beta-2 AR gene to blood pressure regulation, heart size, and probably to the development of hypertension. We suggest that a combined linkage and association approach will elucidate the genetic variability influencing blood pressure and other cardiovascular phenotypes. PMID- 10679498 TI - Hypertension incidence is predicted by high levels of hopelessness in Finnish men. AB - Recent studies have reported that hopelessness is an important factor in cardiovascular morbidity and mortality, including ischemic heart disease, acute myocardial infarction, and atherosclerotic progression. This study examined the relationship between hopelessness and incident hypertension in a population-based sample of 616 initially normotensive, middle-aged men from eastern Finland, an area with high rates of cardiovascular disease. Participants completed a medical examination and a series of psychological questionnaires at baseline and at the 4 year follow-up. Hopelessness was measured by 2 items assessing negative expectancy about the future and one's goals. A logistic regression model with adjustments for age, body mass index, baseline resting blood pressure, physical activity, smoking, alcohol consumption, education, parental history of hypertension, and self-reported depressive symptoms revealed that men reporting high levels of hopelessness at baseline were 3 times more likely to become hypertensive (systolic blood pressure > or =165 mm Hg and/or a diastolic blood pressure > or =95 mm Hg or confirmed use of antihypertensive medication) in the intervening 4 years than men who were not hopeless (odds ratio, 3.22; 95% confidence interval, 1. 56, 6.67). Men reporting moderate levels of hopelessness were not at a significantly increased risk of hypertension (odds ratio, 1.27; 95% confidence interval, 0.79, 2.07). This is the first study to identify a significant relationship between hopelessness and incident hypertension. Research is needed to explore the neuroendocrine and central nervous system mechanisms underlying this association. PMID- 10679499 TI - Contribution of autosomal loci and the Y chromosome to the stress response in rats. AB - Stress is a critical contributor to cardiovascular diseases through its impact on blood pressure variability and cardiac function. Familial clustering of reactivity to stress has been demonstrated in human subjects, and some rodent models of hypertension are hyperresponsive to stress. Therefore, the present study was designed to uncover the genetic determinants of the stress response. We performed a total genome linkage search to identify the loci of the body temperature response to immobilization stress in a set of recombinant inbred strains (RIS) originating from reciprocal crosses of spontaneously hypertensive rats (SHR) with a normotensive Brown Norway Lx strain. Two quantitative trait loci (QTLs) were revealed on chromosomes (Chrs) 10 and 12 (logarithm of odds scores, 2.2 and 1. 3, respectively). The effects of these QTLs were enhanced by a high sodium diet (logarithm of odds scores, 4.0 and 3.3 for Chrs 10 and 12, respectively), which is suggestive of a salt-sensitive component for the phenotype. Congenics for Chr 10 confirmed both the QTL and the salt effect in RIS. Negatively associated loci were also identified on Chrs 8 and 11. Interaction between the loci of Chrs 10 and 12 was demonstrated, with the rat strains bearing SHR alleles at both loci having the highest thermal response to stress. Furthermore, the Y Chr of SHR origin enhanced the response to immobilization stress, as demonstrated in 2 independent models, RIS and Y Chr consomics. However, its full effect requires autosomes of the SHR strain. These findings provide the first evidence for the genetic determination of reactivity to stress with interactions between autosomal loci and between the Y and autosomal Chrs that contribute to the explanation of the 46% of variance in the stress response. PMID- 10679500 TI - Heritability of central systolic pressure augmentation: a twin study. AB - Less than 50% of the variance in left ventricular mass is explained by conventional factors such as age, blood pressure, and body size. Genetic influences may account for part of the unexplained variance. The central (aortic) pressure augmentation index has been suggested as a noninvasive measure of pulsatile load, which is a likely determinant of left ventricular mass. We quantified the genetic influence on augmentation index and determined the extent to which this influence is dependent on the effects of age, height, heart rate, and blood pressure. We performed a classical twin study composed of 225 monozygotic and 594 dizygotic female white twin pairs aged 18 to 73 years. Augmentation index and mean arterial pressure were based on the central pressure wave derived from the radial waveform as measured by applanation tonometry. Quantitative genetic modeling techniques were used to analyze the data. The heritability of augmentation index was 37%, whereas heritabilities for blood pressure traits varied between 13% and 25%. Most of the variance in augmentation index could be explained by genetic and environmental factors specifically influencing augmentation index. Only a relatively small part of the total variance in augmentation index could be attributed to genes in common with height (3.1%), heart rate (4.6%), and mean arterial pressure (5.6%). Age explained 19% of the total variation in augmentation index. In conclusion, augmentation index has a significant heritable component, which is largely independent of the influence of blood pressure, heart rate, height, and age. Finding genes for the augmentation index could help to unravel pathophysiological mechanisms causing left ventricular hypertrophy and lead to improvements in prevention, diagnosis, and treatment of at-risk populations. PMID- 10679501 TI - Continuous relation between left ventricular mass and cardiovascular risk in essential hypertension. AB - The detection of left ventricular (LV) hypertrophy on echocardiography is a powerful risk indicator in essential hypertension. However, the prognostic impact of LV mass values within the "normal" range and the shape of the relation between LV mass and prognosis remain unclear. Thus, 1925 white subjects with uncomplicated essential hypertension underwent off-therapy 24-hour blood pressure monitoring and M-mode echocardiography. During 4. 0+/-2 years of follow-up, there were 181 major cardiovascular events (2.4/100 patient-years) and 49 deaths from all causes. In the 5 gender-specific quintiles of LV mass distribution (partition values: 92, 105, 120, and 138 g/m(2) in men and 79, 91, 102, and 116 g/m(2) in women), cardiovascular event rates were 0.8, 1.7, 2.2, 2.9, and 4. 3 per 100 patient-years. After adjustment for several risk factors, including 24-hour ambulatory blood pressure, the relative risk (RR) of developing a cardiovascular event increased progressively from the first quintile (RR 1) to the second (RR 1.6, 95% CI 0.8 to 3.1), third (RR 1.9, 95% CI 1.01 to 4.0), fourth (RR 3.0, 95% CI 1.5 to 5. 8), and fifth (RR 3.5, 95% CI 1.8 to 6.8) quintile. For all-cause death, the RR in the fifth quintile compared with the first quintile was 4.3 (95% CI 1.2 to 13.4). In conclusion, the powerful relation between LV mass and risk of cardiovascular disease in subjects with uncomplicated essential hypertension is continuous over a wide range of LV mass values, even below the current "upper normal" limits. The relation remains significant after control for traditional risk factors, including ambulatory blood pressure. PMID- 10679502 TI - Blood pressure-independent effects in rats with human renin and angiotensinogen genes. AB - The blood pressure-independent effects of angiotensin II (Ang II) were examined in double transgenic rats (dTGR) harboring human renin and human angiotensinogen genes, in which the end-organ damage is due to the human components of the renin angiotensin system. Triple-drug therapy (hydralazine 80 mg/L, reserpine 5 mg/L, and hydrochlorothiazide 25 mg/L in drinking water) was started immediately after weaning. Triple-drug therapy normalized blood pressure and coronary resistance, only partially prevented cardiac hypertrophy, and had no effect on ratio of renal weight to body weight. Although triple-drug therapy delayed the onset of renal damage, severe albuminuria nevertheless occurred. Semiquantitative scoring of ED 1-positive and MIB-5-positive (nuclear cell proliferation-associated antigen Ki 67) cells showed profound perivascular monocyte/macrophage infiltration and cell proliferation in kidneys and hearts of untreated dTGR. Triple-drug therapy had only a minimal effect on local inflammatory response or vascular cell proliferation. In contrast, a novel orally active human renin inhibitor (HRI), 30 mg/kg by gavage for 4 weeks, normalized blood pressure and coronary resistance and also prevented cardiac hypertrophy and albuminuria. ED-1-positive cells and MIB-5-positive cells were decreased by HRI in hearts and kidneys almost to levels observed in normotensive Sprague-Dawley rats. The renoprotective effects of HRI were at least in part due to improved renal hemodynamics and distal tubular function, since HRI shifted renal pressure-diuresis/natriuresis curves leftward by approximately 35 mm Hg, increased glomerular filtration rate and renal blood flow, and shifted the fractional water and sodium excretion curves leftward. In untreated dTGR, plasma Ang II was increased by 400% and renal Ang II level was increased by 300% compared with Sprague-Dawley rats. HRI decreased plasma human renin activity by 95% and normalized Ang II levels in both plasma and kidney compared with triple-drug therapy. Our findings indicate that in dTGR harboring human renin and angiotensinogen genes, Ang II causes end-organ damage and promotes inflammatory response and cellular growth largely independent of blood pressure. PMID- 10679503 TI - Gene transfer of endothelial nitric oxide synthase reduces angiotensin II-induced endothelial dysfunction. AB - Angiotensin II stimulates vascular NADPH oxidase to produce superoxide, which can react with nitric oxide and impair vasomotor function. We tested the hypothesis that the overexpression of endothelial nitric oxide synthase (eNOS) or superoxide dismutase (SOD) would correct angiotensin II-induced endothelial dysfunction. We examined the effects of the gene transfer of eNOS or 2 isoforms of SOD to the aorta in angiotensin II-treated rabbits on vasomotor function. New Zealand White rabbits were treated for 1 week with angiotensin II (100 ng. kg(-1). min(-1)) or saline by osmotic minipumps. In angiotensin II-treated rabbits, mean blood pressure was 107+/-8 mm Hg; it was 67+/-5 mm Hg in saline-infused rabbits (P<0.05). In aortas from angiotensin II-treated rabbits, lucigenin-enhanced chemiluminescence demonstrated a 2.5-fold increase in superoxide levels, and the oxidative fluorescent probe hydroethidine indicated increased superoxide levels throughout the vascular wall, especially in the endothelium and adventitia. Maximal relaxation to acetylcholine was less in aortas from rabbits treated with angiotensin II (72+/-5% versus 87+/-4% in saline-treated rabbits; P<0.01), but responses to sodium nitroprusside were similar. Segments of the thoracic aorta were incubated in vitro with an adenoviral vector that expressed eNOS, copper zinc SOD (CuZnSOD), extracellular SOD (ECSOD), or beta-galactosidase. beta-Gal treatment with adenovirus containing the gene for eNOS (AdeNOS) but not adenovirus containing the gene for beta-gal (Adbeta-gal) (control virus) restored responses to acetylcholine (82+/-3% after AdeNOS and 67+/-4% after Adbeta-gal). Gene transfer of CuZnSOD or ECSOD did not improve the endothelium-dependent relaxation of the aorta in rabbits that received angiotensin II. Thus, gene transfer of eNOS, but not SOD, effectively restores vasomotor function in angiotensin II-infused rabbits. PMID- 10679504 TI - Vasopressin does not effect hypertension caused by long-term nitric oxide inhibition. AB - Nitric oxide attenuates both vasopressin-induced vasoconstriction and vasopressin release. We tested whether hypertension and renal dysfunction elicited by chronic inhibition of nitric oxide (NO) synthesis using N(G)-nitro-L-arginine (L-NNA) could be mediated in part by vasopressin V(1A) receptors. Male rats were treated orally for 6 weeks with L-NNA (15 mg/kg per day), a nonpeptide V(1A) receptor antagonist (2S)-1-[(2R,3S)-5-chloro-3-(2-chlorophenyl)-1-(3, 4-dimethoxybenzene sulfonyl)-3-hydroxy-2, 3-dihydro-1H-indole-2-carbonyl]-pyrrolidine-2-carboxamide (SR 49059, 30 mg/kg per day), or a combination of SR 49059 and L-NNA (same doses), or they received no treatment. Both drugs were added to the food. Measurements were performed in conscious rats (urine collection in metabolic cages, tail-cuff arterial pressure) and at the end of the study in anesthetized rats (clearance measurements). L-NNA produced sustained hypertension, decreased glomerular filtration rate, and increased renal vascular resistance, plasma renin activity, and urinary albumin excretion. SR 49059 had no effect per se on these parameters and also did not attenuate the hypertension and renal dysfunction induced by L-NNA. Surprisingly, SR 49059 potentiated L-NNA-induced hypertension at the end of the 6-week treatment. However, the blood pressure response and the renal and mesenteric vasoconstriction elicited by exogenous vasopressin were attenuated in rats treated with SR 49059. L-NNA did not change plasma vasopressin concentration or 24-hour urinary vasopressin excretion. Our findings suggest that activation of vasopressin V(1A) receptors does not contribute to the hypertension and renal dysfunction induced by chronic NO synthesis inhibition. They also document unchanged plasma vasopressin concentration in NO-deficient hypertension. PMID- 10679505 TI - Role of alpha(2)-adrenergic receptor subtypes in the acute hypertensive response to hypertonic saline infusion in anephric mice. AB - Experimental evidence suggests that the acute hypertensive response induced in anephric animals by infusion of a hypertonic saline solution is mediated by disinhibition of the presynaptic sympathoinhibitory alpha(2)-adrenergic receptors (alpha(2)-AR) of the central nervous system. The purpose of the present experiments was to dissect the role of the 3 distinct alpha(2)-AR subtypes (alpha(2A)-, alpha(2B), - and alpha(2C)-AR) in this response. Groups of genetically engineered mice deficient in each one of these alpha(2)-AR subtype genes were submitted to bilateral nephrectomy followed by a 0.4-mL infusion of 4% saline over a 2-hour period, with constant direct blood pressure (BP) monitoring. The alpha(2A)-AR-deficient and alpha(2C)-AR-deficient mice responded with significant BP elevations (by 11.8+/-2.5 and 16.7+/-1.7 mm Hg, respectively), and so did their wild-type counterparts (17.8+/-2.5 and 11.8+/-2.0 mm Hg, respectively) and the wild-type alpha(2B) +/+ (13.1+/-2.4 mm Hg). However, the alpha(2B)-AR-deficient mice were unable to raise their BP and had a slightly lowered BP (by -3.0+/-4. 0 mm Hg) at the end of the infusion period. All 6 groups exhibited elevated plasma norepinephrine levels ranging between 0.8 and 1.8 ng/mL at the end of the infusion. In all cases, the alpha(2)-AR-deficient groups tended to have higher norepinephrine levels than their wild-type counterparts. Surprisingly, this difference was significant only in the alpha(2B)-AR-deficient mice, which, despite the elevated norepinephrine, were unable to raise their BP. The data suggest that a full complement of the alpha(2B)-AR is needed to mediate the hypertensive response to acute saline load, even though its absence does not prevent the release of norepinephrine under these conditions. PMID- 10679506 TI - Attenuation of the "white-coat effect" by antihypertensive treatment and regression of target organ damage. AB - This study assessed whether 2 common surrogate measures of the "white-coat effect," namely the clinic-daytime and the clinic-home differences in blood pressure (BP), were attenuated by long-term antihypertensive treatment and whether this attenuation is relevant to the treatment-induced regression of left ventricular hypertrophy, thus having clinical significance. We considered data from 206 patients with essential hypertension (aged 20 to 65 years) who had a diastolic BP between 95 and 115 mm Hg and echocardiographic evidence of left ventricular hypertrophy. In each patient, clinic BP, 24-hour ambulatory BP, and left ventricular mass index were assessed at baseline, after 3 and 12 months of treatment with an angiotensin-converting enzyme inhibitor, and after a final 4 week placebo run-off period. At baseline, the clinic-daytime differences in systolic and diastolic BP were 12.1+/-15.4 and 6.8+/-10.1 mm Hg, respectively; the corresponding values for the clinic-home differences were 5.7+/-10.6 and 2.9+/-6.1 mm Hg, respectively. These differences were reduced by 57.6% and 77.1% (P<0.01) and by 65.7% and 64.3% (P<0.01), respectively, after 12 months of treatment, with a partial return toward the pretreatment differences after the final placebo period. The observed treatment-induced reductions in left ventricular mass index and those in the clinic-daytime or clinic-home differences for systolic and diastolic BP showed no significant relationship when tested by multiple regression analysis. This provides the first longitudinal evidence that clinic-daytime and clinic-home differences in BP have no substantial value in predicting the regression of target organ damage, such as left ventricular hypertrophy, that has prognostic relevance. PMID- 10679507 TI - Amlodipine, enalapril, and dependent leg edema in essential hypertension. AB - Calcium channel blockers (CCBs) blunt postural skin vasoconstriction, an autoregulatory mechanism that minimizes gravitational increases in capillary pressure and avoids fluid extravasation when standing. To evaluate the dose response relation between this pharmacological interference and dependent edema, a frequent side effect of CCBs during antihypertensive treatment, skin blood flow (laser Doppler flowmetry) at the dorsum of the foot, both supine and with the limb passively placed 50 cm below the heart level, and leg weight (Archimedes principle) were measured at baseline, during increasing doses of the dihydropyridine amlodipine (5 and 10 mg UID each for 2 weeks), and after drug withdrawal in 10 hypertensive men. Because angiotensin-converting enzyme inhibitors may attenuate ankle swelling by CCBs, those parameters were evaluated according to a similar design during amlodipine (10 mg UID) and enalapril (20 mg UID) combined (n=10). As a control, the effect of enalapril monotherapy (10 and 20 mg UID for 2 weeks each) was evaluated in a third series of patients (n=8). Amlodipine (5 mg UID) increased leg weight without modifying postural vasoconstriction (the percent skin blood flow decrease from horizontal to dependent position), which indicates that extravascular fluid shift was independent of postural skin vasoconstriction. At 10 mg UID, however, amlodipine blunted postural vasoconstriction and increased leg weight further, which suggests that skin blood flow autoregulation limited additional fluid transfer. Both parameters normalized after drug withdrawal. Enalapril per se did not affect cutaneous vasomotion or leg weight but reduced the amount of dependent fluid extravasation by the CCB despite a persistent antagonism for postural vasoconstrictor responses. PMID- 10679508 TI - A novel stable inhibitor of endopeptidases EC 3.4.24.15 and 3.4.24.16 potentiates bradykinin-induced hypotension. AB - We have developed a novel inhibitor of the metalloendopeptidases EC 3.4.24.15 (EP24.15) and EC 3.4.24.16 (EP24.16), N-[1-(R, S)-carboxy-3-phenylpropyl]-Ala-Aib Tyr-p-aminobenzoate (JA2), in which alpha-aminoisobutyric acid (Aib) is substituted for an alanine in a well-described but unstable inhibitor, cFP-AAY pAB. This substitution increases the resistance of the inhibitor to degradation without altering potency. In the present study, we investigated the effects of JA2 (5 mg/kg) on the responses of mean arterial pressure to bradykinin, angiotensin I, and angiotensin II in conscious rabbits. The depressor responses to both low (10 ng/kg) and high (100 ng/kg) doses of bradykinin were increased 7.0+/-2. 7-fold and 1.5+/-0.3-fold, respectively, during the 30 minutes after JA2 administration (mean+/-SEM, n=8). Bradykinin potentiation was undiminished 4 hours after JA2 injection. In contrast, the hypertensive effects of angiotensins I and II were unaltered, indicating that the bradykinin-potentiating effects were not due to angiotensin-converting enzyme inhibition. These data suggest that JA2 is not only a potent and specific inhibitor of EP24.15 and EP24. 16 but is also stable in vivo. Furthermore, the potentiation of bradykinin-induced hypotension by JA2 suggests for the first time a role for one or both of these peptidases in the metabolism of bradykinin in the circulation. PMID- 10679509 TI - Sympathoinhibition by central and peripheral infusion of nifedipine in spontaneously hypertensive rats. AB - The present study assessed whether central mechanisms may contribute to the hypotensive effect of the calcium channel blocker nifedipine. In conscious, spontaneously hypertensive rats (SHR) on a high-salt diet, hemodynamic (mean arterial pressure [MAP] and heart rate) and sympathetic (renal sympathetic nerve activity) responses to low, central, intracerebroventricular infusion rates (25 microg. kg(-1). h(-1) for 2 hours) and peripheral intravenous rates (50 microg. kg(-1). h(-1) for 3 hours and then 100 microg. kg(-1). h(-1) for 2 hours) of nifedipine were evaluated. The distribution of nifedipine in the blood and tissues was assessed at the end of the infusions. Nifedipine significantly inhibited renal sympathetic nerve activity and lowered MAP in SHR beginning 30 minutes after the start of the intracerebroventricular infusion. The decrease of MAP by intravenous infusion began at 60 minutes and was more profound with 100 microg. kg(-1). h(-1). Inhibition of sympathetic activity preceded and then paralleled the decrease in blood pressure; it occurred earlier with central (15 to 30 minutes) than with peripheral (30 to 60 minutes) infusion. Intravenous infusion resulted in concentrations of nifedipine in brain structures (brain stem, midbrain, and cortex) that were 30% to 40% of those in the heart, kidneys, and liver. From the hemodynamic and sympathetic responses and the distribution of nifedipine into the central nervous system, we conclude that the peripheral infusion of nifedipine at relatively low rates may evoke a hypotensive response in SHR, not only via peripheral mechanisms, but also through central mechanisms, which will lead to an inhibition of sympathetic outflow and, therefore, a lowering of blood pressure. PMID- 10679510 TI - Effect of age on brachial artery wall properties differs from the aorta and is gender dependent: a population study. AB - Compliance and distensibility are wall properties of large arteries, which may play a role in cardiovascular disease. The purpose of this study was to investigate whether the influence of age on these vessel wall properties differs between vascular territories and is gender-dependent. In a population sample of 498 men and women 20 to 79 years of age, diameter, distensibility, and compliance coefficient of the muscular brachial artery were measured with an echo-tracking device. Distensibility of the aorta was measured with the use of pulse-wave velocity. The effects of age and gender were assessed and adjusted for confounding factors such as mean blood pressure, pulse rate, body mass index, smoking, alcohol intake, and antihypertensive treatment. Covariance analysis showed no relation between gender and distensibility of the elastic aorta. Distensibility of the muscular brachial artery was lower in men, whereas men had a larger diameter and larger compliance of the brachial artery. With age, distensibility of the aorta decreased in both sexes to the same extent, whereas distensibility of the brachial artery did not change significantly. With age, brachial artery diameter increased; this increase was more pronounced in women. In men brachial artery compliance did not change with age, whereas in women compliance of the brachial artery increased with age. This study (1) confirms that distensibility of the aorta, an elastic artery, decreases with age. (2) In contrast to the aorta, after adjustment for confounding factors, in both men and women, no relation exists between age and distensibility of the muscular brachial artery. (3) Brachial artery diameter increase with age is more pronounced in women than in men. (4) In contrast to the well-known decrease in arterial compliance of elastic arteries with age, brachial artery compliance is not decreased with age and is increased in women. In conclusion, the effect of age on large-artery wall properties is not uniform but depends on gender and vascular territory. PMID- 10679511 TI - Age-related changes in renal cyclic nucleotides and eicosanoids in response to sodium intake. AB - The signaling molecules cGMP, cAMP, prostaglandin E(2) (PGE(2)), and prostaglandin F(2alpha) (PGF(2alpha)) play important roles in mediating the response of the kidney to changes in dietary sodium intake. We used a renal microdialysis technique in conscious rats to address the hypothesis that the renal ability to produce these mediators in response to dietary sodium intake is altered during maturation. Young (4-week-old) or adult (6-month-old) rats were studied after the consumption for 5 days of diets containing low (0. 04% NaCl), normal (0.28% NaCl), or high (4.0% NaCl) levels of sodium. Plasma renin activity was significantly increased by low-sodium diet and significantly decreased by high-sodium diet, with no significant difference between the responses of the 2 age groups. Renal interstitial fluid (RIF) levels of cGMP, cAMP, PGE(2), and PGF(2alpha) on normal-sodium diet were similar in the 2 age groups. Low-sodium diet caused a significant increase in RIF levels of all 4 mediators, with no significant differences between the responses of the 2 age groups. High-sodium diet also caused a significant increase in RIF levels of all 4 mediators. However, RIF production of cGMP, cAMP, and PGE(2) was significantly greater, and RIF PGF(2alpha) production was significantly lower, in young rats compared with adult rats. These data demonstrate that the kidneys of young and adult rats respond to dietary sodium restriction in a similar manner but that there are age related changes in the renal response to sodium loading. PMID- 10679512 TI - Elevated perfusion pressure upregulates endothelin-1 and endothelin B receptor expression in the rabbit carotid artery. AB - To investigate the hypothesis that high blood pressure activates the endothelin system in the vessel wall, isolated segments of the rabbit carotid artery were subjected to different levels of perfusion pressure. Both preproendothelin-1 (ppET-1) mRNA abundance and intravascular ET-1 peptide content were strongly upregulated on raising the intraluminal pressure from 90 to 160 mm Hg for 3 to 12 hours, and this increase in ppET-1 mRNA occurred predominantly in the endothelial cells. Endothelin-converting enzyme-1 and endothelin A receptor (ET(A)-R) expression were pressure-insensitive, whereas that of the ET(B)-R in the smooth muscle cells was also significantly enhanced. Both the pressure-induced increase in ppET-1 and ET(B)-R expression required RNA synthesis because they were abolished by actinomycin D. The nuclear signaling mechanisms involved therein, however, appeared to be different. Thus, the pressure-induced expression of ppET 1 and activation of CCAAT-enhancer binding proteins beta and delta were blocked by the tyrosine kinase inhibitor herbimycin A, whereas ET(B)-R expression and the nuclear translocation of activator protein-1 were abolished by the protein kinase C inhibitor Ro 31-8220. One consequence of these presumably deformation-induced changes in gene expression was an increased rate of apoptosis of the smooth muscle cells in the media that if transferable to the situation in human blood vessels may contribute to hypertension-induced arterial remodeling. PMID- 10679513 TI - Effects of systemic inhibition of neuronal nitric oxide synthase in diabetic rats. AB - Diabetes is associated with alterations in nitric oxide-mediated vasomotor function. The role of nitric oxide generated via the neuronal nitric oxide synthase pathway in the control of systemic and renal hemodynamics in diabetes has not been studied. To explore the hypothesis that diabetic vascular dysfunction is in part caused by altered neuronal nitric oxide synthase activity, systemic and renal hemodynamics were assessed before and after acute inhibition of this enzyme with a specific inhibitor, S-methyl-L-thiocitrulline, in control and diabetic rats. The interaction of this pathway and the renin-angiotensin system was studied in separate groups of rats pretreated with the angiotensin II receptor blocker losartan; these rats were compared with rats treated with losartan alone. Diabetic animals demonstrated higher baseline glomerular filtration rates and filtration fractions. At a low dose, the neuronal nitric oxide synthase inhibitor induced similar dose-dependent pressor responses in control and diabetic rats. Losartan abolished the pressor response in both groups. No changes in renal plasma flow or renal vascular resistance occurred in control rats. In contrast, diabetic rats responded with significant renal vasoconstriction. At a high dose, the renal vasoconstriction was similar in both groups and was not affected by losartan. In conclusion, neuronal nitric oxide synthase-derived nitric oxide plays a role in the control of systemic and renal hemodynamics in normal and diabetic rats. Diabetic rats are more sensitive to the inhibitor, suggesting increased activity of this pathway in the diabetic kidney. Furthermore, renal responses in diabetic rats were attenuated by angiotensin II receptor blockade, whereas losartan alone induced hemodynamic changes that were opposite those seen with neuronal nitric oxide synthase inhibition. This observation implicates angiotensin II as an important modulator of this nitric oxide pathway in diabetes. PMID- 10679514 TI - Blood pressure is related to placental volume and birth weight. AB - The objective of this study was to determine whether maternal nutrition and fetal and placental size program blood pressure. A longitudinal study linking the maternal anthropometric measurements of the first antenatal visit, ultrasound data of placental and fetal size, anthropometry at birth, and childhood growth and blood pressure was performed. The subjects were 428 women who attended the antenatal clinic at the University Hospital of the West Indies, Kingston, Jamaica, and their children, who were subsequently followed up. Systolic blood pressure at ages 1, 2, 2.5, 3, and 3.5 years was the main outcome measure. Pooling the data across ages, systolic blood pressure fell by 1.4 mm Hg for every 1-kg increase in birth weight (95% CI 0.2 to 2.7, P=0.02) and by 1.2 mm Hg for every 100-mL increase in placental volume at 20 weeks of gestation (95% CI 0.4 to 2.0, P=0.004). Blood pressure was also negatively associated with placental volume at 17 weeks and fetal abdominal circumference at 20 weeks. Measures of maternal nutritional status were strongly related to birth weight and placental volume but not directly to childhood blood pressure at these young ages. In conclusion, blood pressure is associated with fetal size in this population, as previously described among Europeans. We found associations between placental volume and abdominal circumference in the second trimester and childhood blood pressure, suggesting that the initiating events of blood pressure programming occur early in pregnancy. Measures of maternal nutritional status were not directly related to childhood blood pressure at these young ages but were strong predictors of both birth weight and placental volume, suggesting an indirect relation. PMID- 10679515 TI - Glucocorticoid-remediable aldosteronism and pregnancy. AB - Glucocorticoid-remediable aldosteronism (GRA) is a hereditary form of primary hyperaldosteronism that presents with hypokalemia and hypertension from childhood onward. GRA is characterized by the ectopic production of aldosterone in the cortisol-producing zona fasciculata under the regulation of adrenocorticotrophic hormone. Despite the early age of onset, no previous reports of pregnancy and GRA exist. Therefore, we set out to describe the maternal and fetal outcomes of pregnancy in women with GRA. Data regarding the blood pressure and pregnancy outcomes were collected in a retrospective chart review of prenatal and hospital records of 35 pregnancies in 16 women with genetically proven GRA. A total of 6% of pregnancies in women with GRA (GRA+) were complicated by preeclampsia. The published rates of preeclampsia in general obstetric populations vary from 2.5% to 10%. Despite the lack of an apparent increase in the rate of preeclampsia, GRA+ women with chronic hypertension had a high rate (39%) of pregnancy aggravated hypertension. Starting with a higher baseline blood pressure, maternal blood pressure plotted over the time course of pregnancy followed a quadratic curve similar to that previously described in normal pregnancy. Mean gestational age at delivery was 39.1 weeks. Mean birth weight, excluding the 3 sets of twins, was 3219 g. However, infants of GRA+ mothers with pregnancy-aggravated hypertension tended to have lower birth weights than those that did not (3019 g versus 3385 g, respectively; P=0.08). The primary cesarean section rate was 32%, which is approximately double that seen in other general or hypertensive obstetric populations. In summary, GRA+ women did not seem to have an increased risk of preeclampsia. However, GRA+ women with chronic hypertension seem to be at an increased risk for an exacerbation of their hypertension during pregnancy. PMID- 10679516 TI - p38 MAP kinase is required for vasopressin-stimulated HSP27 induction in aortic smooth muscle cells. AB - We previously showed that arginine vasopressin (AVP) stimulates heat shock protein 27 (HSP27) induction through protein kinase C activation in aortic smooth muscle A10 cells. In the present study, we examined whether the mitogen-activated protein (MAP) kinase superfamily is involved in the AVP-stimulated HSP27 induction in A10 cells. AVP stimulated the phosphorylation of p42/p44 MAP kinase and p38 MAP kinase. On the contrary, AVP had little effect on SAPK (stress activated protein kinase)/JNK (c-Jun N-terminal kinase) phosphorylation. The HSP27 accumulation by AVP was not affected by PD98059, an inhibitor of the upstream kinase that activates p42/p44 MAP kinase. SB203580 and PD169316, specific inhibitors of p38 MAP kinase, suppressed the AVP-induced accumulation of HSP27. 12-O-tetradecanoylphorbol 13-acetate, an activator of protein kinase C, induced accumulation of HSP27 and was not inhibited by PD98059 but was inhibited by SB203580. Calphostin C and ET-18-OCH(3), inhibitors of protein kinase C, reduced the phosphorylation of p38 MAP kinase by AVP. SB203580 and PD169316 suppressed the AVP-increased levels in mRNA for HSP27. Dissociation of the aggregated HSP27 to the dissociated HSP27 was induced by AVP. These results strongly suggest that p38 MAP kinase takes part in the pathway of the AVP stimulated induction of HSP27 in vascular smooth muscle cells. PMID- 10679517 TI - Cardiovascular effects of 2-arachidonoyl glycerol in anesthetized mice. AB - Cannabinoids, including the endogenous ligand anandamide, elicit pronounced hypotension and bradycardia through the activation of CB1 cannabinoid receptors. A second endogenous cannabinoid, 2-arachidonoyl glycerol (2-AG), has been proposed to be the natural ligand of CB1 receptors. In the present study, we examined the effects of 2-AG on mean arterial pressure and heart rate in anesthetized mice and assessed the role of CB1 receptors through the use of selective cannabinoid receptor antagonists and CB1 receptor knockout (CB1(-/-)) mice. In control ICR mice, intravenous injections of 2-AG or its isomer 1-AG elicit dose-dependent hypotension and moderate tachycardia that are unaffected by the CB1 receptor antagonist SR141716A. The same dose of SR141716A (6 nmol/g IV) completely blocks the hypotensive effect and attenuates the bradycardic effect of anandamide. 2-AG elicits a similar hypotensive effect, resistant to blockade by either SR141716A or the CB2 antagonist SR144528, in both CB1(-/-) mice and their homozygous (CB1(+/+)) control littermates. In ICR mice, arachidonic acid (AA, 15 nmol/g IV) elicits hypotension and tachycardia, and indomethacin (14 nmol/g IV) inhibits the hypotensive effect of both AA and 2-AG. Synthetic 2-AG incubated with mouse blood is rapidly (<2 minutes) and completely degraded with the parallel appearance of AA, whereas anandamide is stable under the same conditions. A metabolically stable ether analogue of 2-AG causes prolonged hypotension and bradycardia in ICR mice, and both effects are completely blocked by SR141716A, whereas the same dose of 2-AG-ether does not influence blood pressure and heart rate in CB1(-/-) mice. These findings are interpreted to indicate that exogenous 2-AG is rapidly degraded in mouse blood, probably by a lipase, which masks its ability to interact with CB1 receptors. Although the observed cardiovascular effects of 2-AG probably are produced by an arachidonate metabolite through a noncannabinoid mechanism, the CB1 receptor-mediated cardiovascular effects of a stable analogue of 2-AG leaves open the possibility that endogenous 2-AG may elicit cardiovascular effects through CB1 receptors. PMID- 10679518 TI - Vascular response to angiotensin II in atherosclerosis: role of the baroreflex. AB - High-cholesterol alimentation is associated with an induction of angiotensin converting enzyme and angiotensin II receptor expression within the vascular wall of the aorta. Despite an enhanced pressure response to angiotensin II in atherosclerotic conscious rabbits, angiotensin II-induced contraction was reduced in isolated vascular rings from the aorta and unchanged in those from the iliac artery. We, therefore, investigated whether cholesterol-induced atherosclerosis enhances overall vascular responsiveness to angiotensin II in intact animals and whether an altered arterial baroreflex sensitivity can explain the discrepancy between experiments in intact animals and isolated blood vessels. Rabbits were maintained on a high-cholesterol diet (2 g/d cholesterol plus 20 mL/d sunflower seed oil, n=11) or on a standard diet (n=12) for 12 weeks. Total serum lipids markedly increased (P<0.05). Tissue examinations 6 weeks after termination of the high-cholesterol diet revealed distinct atherosclerosis and elevated cholesterol content in the aorta (P<0.05). A high-cholesterol diet did not change baseline hemodynamic parameters. However, angiotensin II-induced increases in total peripheral resistance were larger in the atherosclerotic animals (86.3+/-13.0 versus 41.9+/-9.7 mm Hg. L(-1). min, P<0.05). In addition, the blood pressure pulse interval relationship was markedly reduced (slope: 0.80+/-0.14 versus 0. 49+/-0.06 ms/mm Hg, P<0.05), which suggested that the baroreflex blunted the angiotensin II response to a lesser extent in atherosclerotic animals. In conclusion, the overall vascular responsiveness to angiotensin II is increased in the atherosclerotic rabbit as indicated by the larger increase in total peripheral resistance. An attenuation of the arterial baroreflex sensitivity may contribute to this effect. PMID- 10679519 TI - Hypertension online only : february 2000 PMID- 10679520 TI - Long-term telemetric recording of arterial pressure and heart rate in mice fed basal and high NaCl diets. AB - Research examining the control of arterial pressure in mice has primarily relied on tail-cuff plethysmography and, more recently, on tethered arterial catheters. In contrast, the radiotelemetry method has largely become the "gold standard" for long-term monitoring of arterial pressure and heart rate in rats. Whereas smaller telemetry probes have recently been developed, no published studies have used radiotelemetric monitoring of arterial pressure in mice, largely because of a relatively low success rate in small mice (ie, <30 g body weight). We report on the development of a protocol for the use of these probes to continuously monitor arterial pressure and heart rate in mice as small as 19 g body weight. To test the accuracy and reliability of this method, adult C57/BL6 mice were monitored for 3 weeks during exposure to a basal followed by a high NaCl diet. The results demonstrate that carotid and aortic placements of the telemetry probe provide equally accurate monitoring of arterial pressure and heart rate, but the carotid placement has a much greater rate of success. Exposure to a high NaCl diet increases both the amplitude of the arterial pressure rhythm (+ 6.0+/-0.6 mm Hg, approximately 32%) and the average mean arterial pressure (+ 8.6+/-1.1 mm Hg, approximately 8%), as would be predicted from previous studies in NaCl-resistant rats. Thus, the data demonstrate that telemetric recording of long-term arterial pressure and heart rate provides a powerful tool with which to define the mechanisms of cardiovascular control in mice. PMID- 10679521 TI - Heart rate variability: how to assess effects of mild therapies on autonomic control in small groups of mild and borderline hypertensives? PMID- 10679522 TI - Stability of the molten globule state of a domain-exchanged chimeric protein between human and bovine alpha-lactalbumins. AB - A domain-exchanged chimeric alpha-lactalbumin (alpha-LA), which consisted of the alpha-domain of human alpha-LA and the beta-domain of bovine alpha-LA, was constructed. Like native alpha-LA, the chimeric protein was in a molten globule state in the absence of Ca(2+) at neutral pH and low salt concentration. The stability of the molten globule state of the constructed chimeric protein was identical to that of the recombinant human protein and was higher than that of the recombinant bovine protein. The stability of the molten globule state of alpha-LA is defined by the stability of the alpha-domain. PMID- 10679523 TI - Production of an activated form of Bacillus stearothermophilus L-2-hydroxyacid dehydrogenase by directed evolution. AB - Bacillus stearothermophillus lactate dehydrogenase (bsLDH) is activated in the presence of fructose 1,6 bisphosphate (FBP). The activator is expensive and representative of the sort of co-factor complications that are undesirable in industrial processes. Three rounds of random mutagenesis and screening produced a mutant (6A) which is almost fully activated in the absence of FBP. Wild-type bsLDH has a K(pyr)(M) of 5 mM in the absence of FBP but when activated (+FBP) the K(pyr)(M) drops to 0.05 mM. The mutant 6A has a K(pyr)(M) of 0.07 mM in the absence of FBP. 6A has three amino acid substitutions-R118C, Q203L and N307S resulting in a 70-fold activation, none of the mutations are near the active site. The activation of wild type bsLDH is due to an FBP induced tetramerization of dimeric bsLDH bringing about a structural rearrangement of key active site residues. The most likely explanation for the activation of 6A is derived from the position of Q203L, which is at the dimer-dimer interface. The suggestion is that the hydrophilic to hydrophobic change has altered the dimer-tetramer equilibrium position towards that of the tetramer. What is significant is the activation of bsLDH by a subtle long range event produced by the 'blind' directed evolution approach. PMID- 10679524 TI - Protein thermal stability: insights from atomic displacement parameters (B values). AB - The factors contributing to the thermal stability of proteins from thermophilic origins are matters of intense debate and investigation. Thermophilic proteins are thought to possess better packed interiors than their mesophilic counterparts, leading to lesser overall flexibility and a corresponding reduction in surface-to-volume ratio. These observations prompted an analysis of B values reported in high-resolution X-ray crystal structures of mesophilic and thermophilic proteins. In this analysis, the following aspects were addressed: (1) frequency distribution of normalized B values (B' factors) over all the proteins and for individual amino acids; (2) amino acid compositions in high B value regions of polypeptide chains; (3) variation in the B values from core to the surface of proteins in terms of their radius of gyration; and (4) degree of dispersion of normalized B values in spheres around the Calpha atoms. The analysis revealed that (1) Ser and Thr have lesser flexibility in thermophiles than in mesophiles, (2) the proportion of Glu and Lys in high B value regions of thermophiles is higher and that of Ser and Thr is lower and (3) the dispersion of B values within spheres at Calpha atoms is similar in mesophiles and thermophiles. These observations reflect plausible differences in the dynamics of thermophilic and mesophilic proteins and suggest amino acid substitutions that are likely to change thermal stability. PMID- 10679525 TI - Is it better to combine predictions? AB - We have compared the accuracy of the individual protein secondary structure prediction methods: PHD, DSC, NNSSP and Predator against the accuracy obtained by combing the predictions of the methods. A range of ways of combing predictions were tested: voting, biased voting, linear discrimination, neural networks and decision trees. The combined methods that involve 'learning' (the non-voting methods) were trained using a set of 496 non-homologous domains; this dataset was biased as some of the secondary structure prediction methods had used them for training. We used two independent test sets to compare predictions: the first consisted of 17 non-homologous domains from CASP3 (Third Community Wide Experiment on the Critical Assessment of Techniques for Protein Structure Prediction); the second set consisted of 405 domains that were selected in the same way as the training set, and were non-homologous to each other and the training set. On both test datasets the most accurate individual method was NNSSP, then PHD, DSC and the least accurate was Predator; however, it was not possible to conclusively show a significant difference between the individual methods. Comparing the accuracy of the single methods with that obtained by combing predictions it was found that it was better to use a combination of predictions. On both test datasets it was possible to obtain a approximately 3% improvement in accuracy by combing predictions. In most cases the combined methods were statistically significantly better (at P = 0.05 on the CASP3 test set, and P = 0.01 on the EBI test set). On the CASP3 test dataset there was no significant difference in accuracy between any of the combined method of prediction: on the EBI test dataset, linear discrimination and neural networks significantly outperformed voting techniques. We conclude that it is better to combine predictions. PMID- 10679526 TI - Effect of the reaction field electrostatic term on the molecular dynamics simulation of the activation domain of procarboxypeptidase B. AB - Molecular dynamics simulations of the activation domain of porcine procarboxypeptidase B (ADBp) were performed in order to examine the effects of the inclusion of a reaction field (RF) term into the calculation of electrostatics forces for highly charged proteins. Two simulations were performed with the GROMOS96 package, studying the influence of counterions on the final results. Comparison with previous results without the inclusion of the RF term (Marti-Renom, M.A., Mas,J.M., Oliva,B., Querol,E. and Aviles,F.X., Protein Engng, 1998, 11, 101-110) shows that the structure is well maintained when the RF term is included. Moreover, the analysis of the trajectories shows that simulations of solvated highly-charged proteins are sensitive to the presence of counterions, the secondary structures being more stable when their charges are neutralized. PMID- 10679527 TI - Molecular modeling of the collagen-like tail of asymmetric acetylcholinesterase. AB - The asymmetric form of acetylcholinesterase comprises three catalytic tetramers attached to ColQ, a collagen-like tail responsible for the anchorage of the enzyme to the synaptic basal lamina. ColQ is composed of an N-terminal domain which interacts with the catalytic subunits of the enzyme, a central collagen like domain and a C-terminal globular domain. In particular, the collagen-like domain of ColQ contains two heparin-binding domains which interact with heparan sulfate proteoglycans in the basal lamina. A three-dimensional model of the collagen-like domain of the tail of asymmetric acetylcholinesterase was constructed. The model presents an undulated shape that results from the presence of a substitution and an insertion in the Gly-X-Y repeating pattern, as well as from low imino-acid regions. Moreover, this model permits the analysis of interactions between the heparin-binding domains of ColQ and heparin, and could also prove useful in the prediction of interaction domains with other putative basal lamina receptors. PMID- 10679528 TI - An in vitro peptide folding model suggests the presence of the molten globule state during nascent peptide folding. AB - Although molten globules have been widely accepted as a general intermediate in protein folding, there is no clear evidence to show their presence during nascent peptide folding. This paper concentrates on whether the molten globule state occurs, and if it does, when does it form during nascent peptide folding, by comparing the changes in conformation during peptide chain extension of staphylococcal nuclease R. The results show that a large N-terminal fragment of staphylococcal nuclease, SNR121, which already contains more than 80% amino acid sequence of the nuclease, is found to fulfill all the criteria for the molten globule state, suggesting that the molten globule should occur at a later stage of peptide elongation. At this stage the hydrophobic collapse of the polypeptide chain occurs driven by the hydrophobic force, which leads to the formation of a solvent-accessible non-polar core, characterized by the high ANS-binding fluorescence. The nascent peptide folding of the nuclease is a hierarchical process that at the very least includes the following steps: secondary structure accumulation, pre-molten globule state, molten globule state, post-molten globule state and finally the native state. Constant conformation adjustment is necessary for correct folding and active expression of the protein. PMID- 10679529 TI - Functional interdependence of DNA polymerizing and 3'-->5' exonucleolytic activities in Pyrococcus furiosus DNA polymerase I. AB - Pyrococcus furiosus DNA polymerase I (Pol BI) belongs to the family B (alpha like) DNA polymerases and has a strong 3'-->5' exonucleolytic activity, in addition to its DNA polymerizing activity. To understand the relationship between the structure and function of this DNA polymerase, three deletion mutants, Delta1 (DeltaLeu746-Ser775), Delta2 (DeltaLeu717-Ser775) and Delta3 (DeltaHis672 Ser775), and two substituted mutants of Asp405, D405A and D405E, were constructed. These substitutions affected both the DNA polymerizing and the 3'- >5' exonucleolytic activities. The Delta1 mutant protein had DNA polymerizing activity with higher specific activity than that of the wild-type Pol BI, but retained only 10% of the exonucleolytic activity of the wild-type. The other two deletion mutants lost most of both activities. These results suggest that the DNA polymerizing and exonucleolytic activities are closely related to each other in the folded structure of this DNA polymerase, as proposed in the family B DNA polymerases. PMID- 10679530 TI - From DNA sequence to improved functionality: using protein sequence comparisons to rapidly design a thermostable consensus phytase. AB - Naturally-occurring phytases having the required level of thermostability for application in animal feeding have not been found in nature thus far. We decided to de novo construct consensus phytases using primary protein sequence comparisons. A consensus enzyme based on 13 fungal phytase sequences had normal catalytic properties, but showed an unexpected 15-22 degrees C increase in unfolding temperature compared with each of its parents. As a first step towards understanding the molecular basis of increased heat resistance, the crystal structure of consensus phytase was determined and compared with that of Aspergillus niger phytase. Aspergillus niger phytase unfolds at much lower temperatures. In most cases, consensus residues were indeed expected, based on comparisons of both three-dimensional structures, to contribute more to phytase stabilization than non-consensus amino acids. For some consensus amino acids, predicted by structural comparisons to destabilize the protein, mutational analysis was performed. Interestingly, these consensus residues in fact increased the unfolding temperature of the consensus phytase. In summary, for fungal phytases apparently an unexpected direct link between protein sequence conservation and protein stability exists. PMID- 10679531 TI - Tryptophan fluorescence of calmodulin binding domain peptides interacting with calmodulin containing unnatural methionine analogues. AB - The interactions between the abundant methionine residues of the calcium regulatory protein calmodulin (CaM) and several of its binding targets were probed using fluorescence spectroscopy. Tryptophan steady-state fluorescence from peptides encompassing the CaM-binding domains of the target proteins myosin light chain kinase (MLCK), cyclic nucleotide phosphodiesterase (PDE) and caldesmon site A and B (CaD A, CaD B), and the model peptide melittin showed Ca(2+)-dependent blue-shifts in their maximum emission wavelength when complexed with wild-type CaM. Blue-shifts were also observed for complexes in which the CaM methionine residues were replaced by selenomethionine, norleucine and ethionine, and when a quadruple methionine to leucine C-terminal mutant of CaM was studied. Quenching of the tryptophan fluorescence intensity was observed with selenomethionine, but not with norleucine or ethionine substituted protein. Fluorescence quenching studies with added potassium iodide (KI) demonstrate that the non-native proteins limit the solvent accessibility of the Trp in the MLCK peptide to levels close to that of the wild-type CaM-MLCK interaction. Our results show that the methionine residues from CaM are highly sensitive to the target peptide in question, confirming the importance of their role in binding interactions. In addition, we provide evidence that the nature of binding in the CaM-CaD B complex is unique compared with the other complexes studied, as the Trp residue of this peptide remains partially solvent exposed upon binding to CaM. PMID- 10679532 TI - A sparse matrix approach to the solubilization of overexpressed proteins. AB - Many biophysical experiments depend on large amounts of pure, soluble protein. Indeed, the revolution in structural biology has depended on molecular biology's potential to make experiments possible by allowing the overexpression of normally rare proteins in a heterologous host. All too often, however, overexpressed proteins are poorly soluble in buffers that attempt to mimic physiological conditions. Often in such cases the overexpressed protein is assumed to be present in inclusion bodies and hopes of obtaining the desired sample from the overexpression vector are abandoned. We have developed a sparse matrix approach to the solubilization of such proteins that is often successful. This approach relies on well accepted theories of protein solubility and folding to build a sparse matrix that samples 'solubility space' effectively. The buffers of the sparse matrix are used to make crude extracts that are rapidly assayed for soluble protein using gel electrophoresis. We describe our approach and give examples of its application. PMID- 10679533 TI - In what way may race, ethnicity or culture influence asthma outcomes? PMID- 10679534 TI - Asthma education and quality of life in the community: a randomised controlled study to evaluate the impact on white European and Indian subcontinent ethnic groups from socioeconomically deprived areas in Birmingham, UK. AB - BACKGROUND: Whether asthma morbidity in minority groups can be reduced by preventative health care measures delivered in the relevant ethnic dialects requires further evaluation. This study reports clinical outcomes and quality of life from a community based project investigating white European (W/E) and Indian subcontinent (ISC) ethnic groups with asthma living in deprived inner city areas of Birmingham, UK. METHODS: Six hundred and eighty nine asthmatic subjects (345 W/E, 344 ISC) of mean (SD) age 34.5 (15) years (range 11-59) and mean forced expiratory volume in one second (FEV(1)) of 80% predicted were interviewed in English, Punjabi, Hindi, or Urdu. Subjects randomised to the active limb of a prospective, open, randomised, controlled, parallel group, 12 month follow up study underwent individually based asthma education and optimisation of drug therapy with four monthly follow up (active intervention). Control groups were seen only at the beginning and end of the study. Urgent or emergency interactions with primary and secondary health care (clinical outcomes) and both cross sectional and longitudinal data from an Asthma Quality of Life Questionnaire (AQLQ) were analysed. RESULTS: Clinical outcomes were available for 593 subjects. Fewer of the active intervention group consulted their GP (41.8% versus 57.8%, odds ratio (OR) 0.52 (95% CI 0.37 to 0.74)) or were prescribed antibiotics (34.9% versus 51.2%, OR 0.51 (95% CI 0.36 to 0.72)), but by ethnicity statistically significant changes occurred only in the W/E group with fewer also attending A&E departments and requiring urgent home visits. Active intervention reduced the number of hospital admissions (10 versus 30), GP consultations (341 versus 476), prescriptions of rescue oral steroids (92 versus 177), and antibiotics (220 versus 340), but again significant improvements by ethnicity only occurred in the active W/E group. AQLQ scores were negatively skewed to the higher values; regression analysis showed that lower values were associated with ISC ethnicity. Longitudinal changes (for 522 subjects) in the mean AQLQ scores were small but statistically significant for both ethnic groups, with scores improving in the active and worsening in the control groups. CONCLUSIONS: Active intervention only improved clinical outcomes in the W/E group. AQLQ scores, although lower in the ISC group, were improved by active intervention in both ethnic groups. PMID- 10679535 TI - Relationship between exhaled nitric oxide and mucosal eosinophilic inflammation in mild to moderately severe asthma. AB - BACKGROUND: Exhaled levels of nitric oxide (NO) are raised in asthma but the relationship between exhaled NO levels and a direct measure of airway inflammation has not been investigated in asthmatic patients treated with inhaled steroids. METHODS: The relationship between exhaled NO levels, clinical measures of asthma control, and direct markers of airway inflammation were studied in patients with asthma treated with and without inhaled corticosteroids. Thirty two asthmatic patients (16 not using inhaled steroids and 16 using inhaled beclomethasone dipropionate, 400-1000 microg/day) were monitored with respect to measures of asthma control including lung function, symptom scores, medication usage, and variability of peak expiratory flow (PEF) for one month. Measurements of exhaled NO and fibreoptic bronchoscopy were performed at the end of the monitoring period. Bronchial mucosal biopsy specimens were stained with an anti MBP antibody for quantification of eosinophils. RESULTS: There was no significant difference in lung function, symptom scores, or medication usage between the two groups, but there was a significant difference in PEF variability (8.7 (1.2)% in steroid naive patients versus 13.6 (1.9)% in steroid treated patients, p<0.05) and exhaled NO levels (9.9 (3.5) ppb in steroid naive patients versus 13.6 (2.0) ppb in steroid treated patients, p<0.05). There was no correlation between exhaled NO and mucosal eosinophils, or between NO and conventional measures of asthma control. There was a significant correlation between mucosal eosinophils and lung function (r = -0.43, p<0.05). CONCLUSIONS: Exhaled NO levels do not reflect airway mucosal eosinophilia and these markers reflect different aspects of airway inflammation. The clinical usefulness of exhaled NO needs to be determined in prospective longitudinal studies. PMID- 10679536 TI - Is there any relationship between plasma antioxidant capacity and lung function in smokers and in patients with chronic obstructive pulmonary disease? AB - BACKGROUND: It has been suggested that oxidative stress is an important factor in the pathogenesis of chronic obstructive pulmonary disease (COPD). We have shown that an oxidant/antioxidant imbalance occurs in the distal air spaces of smokers and in patients with COPD which is reflected systemically in the plasma. A study was undertaken to determine whether plasma antioxidant status correlated with lung function as assessed by forced expiratory volume in one second (FEV(1)) and forced vital capacity (FVC) in smokers and patients with COPD. METHODS: Plasma antioxidant capacity, assessed by the Trolox equivalent antioxidant capacity (TEAC) as an index of overall systemic oxidative stress, and protein thiol levels were measured in 95 patients with stable COPD, in 82 healthy smokers, and in 37 healthy non-smokers. RESULTS: Mean (SE) plasma TEAC levels were significantly decreased in patients with COPD (0.81 (0.03) mmol/l, p<0.001) and in healthy smokers (0.87 (0.04) mmol/l, p<0. 001) compared with healthy non-smokers (1.31 (0.11) mmol/l). The mean differences in plasma antioxidant capacity (mM) were (0.81, 95% confidence interval (CI) 0.22 to 1.48), (0.87, 95% CI 0.2 to 1.46), and (1.31, 95% CI 1.09 to 1.58) in patients with COPD, healthy smokers, and healthy non-smokers, respectively. This reduction was associated with a 29% (95% CI 18 to 38) and a 30% (95% CI 19 to 40) decrease in plasma protein thiol levels in COPD patients and smokers, respectively. Current smoking was not the main contributor to the reduction in antioxidant capacity in patients with COPD as those patients who were still smokers had similar TEAC levels (mean (SE) 0. 78 (0.05); n = 25) to those who had stopped smoking (0.84 (0.02); n = 70). No significant correlations were found between spirometric data measured as FEV(1) % predicted or FEV(1)/FVC % predicted and the plasma levels of TEAC in patients with COPD, healthy smokers, or healthy non-smokers. Similarly, there was no significant correlation between FEV(1) % predicted or FEV(1)/FVC % predicted and the levels of plasma protein thiols in the three groups. CONCLUSIONS: These data confirm decreased antioxidant capacity in smokers and patients with COPD, indicating the presence of systemic oxidative stress. However, no relationship was found between protein thiols or TEAC levels and measurements of airflow limitation in either smokers or in patients with COPD. PMID- 10679537 TI - Lack of association between ipratropium bromide and mortality in elderly patients with chronic obstructive airway disease. AB - BACKGROUND: Ipratropium is commonly used for the management of elderly patients with obstructive airway disease. However, a recent report suggested that its use might be associated with a significant increase in mortality. A study was therefore conducted to compare all-cause mortality rates between users and non users of ipratropium in elderly patients with either asthma or chronic obstructive pulmonary disease (COPD). METHODS: A retrospective cohort study was performed using linked data from the Canadian Institute for Health Information, the Ontario Drug Benefit Program, the Ontario Health Insurance Plan, and the Ontario Registered Persons database. A total of 32 393 patients were identified who were aged 65 years or older and who had been discharged from hospital with asthma or COPD between 1 April 1992 and 31 March 1997. All-cause mortality rates were compared between those treated and those not treated with ipratropium following discharge from hospital. RESULTS: In total, 49% of patients received ipratropium within 90 days of discharge. After adjusting for age, sex, comorbidity, use of health services, and other airway medications there was no significant association in patients with COPD between the use of ipratropium and mortality (relative risk (RR) 1.03; 95% confidence interval (CI) 0.98 to 1.08). In patients with asthma, however, there was a slight increase in the relative risk of mortality associated with the use of ipratropium (RR 1.24; 95% CI 1.11 to 1.39). A dose-response increase in the mortality rate was not observed with increasing use of ipratropium in either COPD or asthma. CONCLUSIONS: The use of ipratropium in patients with COPD was not associated with an increase in mortality. However, in asthma there was a small increase in the mortality rate. Since asthmatic patients who received ipratropium had greater use of other airway medications and health services, the difference in mortality rate between users and non-users may be a reflection of unmeasured differences in asthma severity. PMID- 10679538 TI - Airflow obstruction in bronchiectasis: correlation between computed tomography features and pulmonary function tests. AB - BACKGROUND: An obstructive defect is usual in bronchiectasis, but the pathophysiological basis of airflow obstruction remains uncertain. High resolution computed tomographic (CT) scanning now allows quantitation of static morphological abnormalities, as well as dynamic changes shown on expiratory CT scans. The aim of this study was to determine which static and dynamic structural abnormalities on the CT scan are associated with airflow obstruction in bronchiectasis. METHODS: The inspiratory and expiratory features on the CT scan of 100 patients with bronchiectasis undergoing concurrent lung function tests were scored semi-quantitatively by three observers. RESULTS: On univariate analysis the extent and severity of bronchiectasis, the severity of bronchial wall thickening, and the extent of decreased attenuation on the expiratory CT scan correlated strongly with the severity of airflow obstruction; the closest relationship was seen between decreased forced expiratory volume in one second (FEV(1)) and the extent of decreased attenuation on the expiratory CT scan (R(s) = -0.55, p<0. 00005). On multivariate analysis bronchial wall thickness and decreased attenuation were consistently the strongest independent determinants of airflow obstruction. The extent of decreased attenuation was positively associated with the severity of bronchial wall thickness, but was not independently linked to gas transfer levels. Endobronchial secretions seen on CT scanning had no functional significance; the severity of bronchial dilatation was negatively associated with airflow obstruction after adjustment for other morphological features. CONCLUSIONS: These findings indicate that airflow obstruction in bronchiectasis is primarily linked to evidence of intrinsic disease of small and medium airways on CT scanning and not to bronchiectatic abnormalities in large airways, emphysema, or retained endobronchial secretions. PMID- 10679539 TI - Exhaled 8-isoprostane as a new non-invasive biomarker of oxidative stress in cystic fibrosis. AB - BACKGROUND: Cystic fibrosis is characterised by oxidative stress in the airways. Isoprostanes are prostaglandin isomers formed by free radical catalysed peroxidation of arachidonic acid. 8-Isoprostane is increased in interstitial lung diseases, asthma, chronic obstructive pulmonary disease, and adult respiratory distress syndrome. Exhaled nitric oxide (NO) and carbon monoxide (CO) are biomarkers of inflammation and oxidative stress in the airways, respectively. METHODS: Concentrations of 8-isoprostane in the breath condensate of 10 normal subjects and 19 patients with stable cystic fibrosis were measured using an enzyme immunoassay (EIA). Breath condensate is a non-invasive method of collecting airway secretions. Exhaled nitric oxide (NO) and carbon monoxide (CO) levels were measured by a chemiluminescence analyser. RESULTS: Concentrations of 8-isoprostane in the breath condensate of patients with stable cystic fibrosis were increased about threefold compared with normal subjects (42.7 (4.5) pg/ml vs 15.2 (1.7) pg/ml; p<0.005, 95% CI 14.6 to 40.9). 8-Isoprostane concentrations were negatively correlated with forced expiratory volume in one second in patients with cystic fibrosis (r = -0.61; p<0.005). Exhaled CO was also increased in patients with cystic fibrosis compared with normal subjects (6.7 (1.2) ppm vs 2.9 (0.3) ppm; p<0.05, 95% CI 0.2 to 7.4). 8-Isoprostane concentrations were significantly correlated with CO levels (r = 0.66; p<0.002). CONCLUSIONS: The results of this study show that oxidative stress is increased in cystic fibrosis and may be quantified by measuring 8-isoprostane concentrations in breath condensate. PMID- 10679540 TI - Management of opportunist mycobacterial infections: Joint Tuberculosis Committee Guidelines 1999. Subcommittee of the Joint Tuberculosis Committee of the British Thoracic Society. PMID- 10679541 TI - Severity prediction rules in community acquired pneumonia: a validation study. AB - BACKGROUND: The British Thoracic Society (BTS) developed a rule (BTSr) based on severity criteria to predict short term mortality in adults admitted to hospital with community acquired pneumonia (CAP). However, neither the BTSr nor a recent modification of it (mBTSr) have been validated in the UK. A case-control study was conducted in a typical UK population to determine the clinical factors predictive of mortality and to assess the performance of these rules. METHODS: Cases were drawn from all patients with CAP who died in 1997 in five large hospitals in the Mid Trent area. Controls were randomly selected from survivors. Factors associated with mortality were identified following review of medical case notes and performance of the severity prediction rules assessed. RESULTS: Age >65 years, temperature <37 degrees C, respiratory rate >24 breaths/min, mental confusion, urea concentration of >7 mmol/l, sodium concentration of <135 mmol/l, and the presence of a pleural effusion, all determined on admission, were independently associated with in-hospital mortality on multivariate analysis. The BTSr was 52% sensitive and 79% specific in predicting death while the mBTSr displayed 66% sensitivity and 73% specificity. CONCLUSIONS: The value of three of the four factors (presence of mental confusion, raised respiratory rate, raised urea) used in the mBTSr as predictors of mortality is confirmed. However, the BTSr and mBTSr did not perform as well in this validation study which included a high proportion (48%) of elderly patients (> or =75 years) compared with the derivation studies. PMID- 10679542 TI - Randomised prospective parallel trial of therapeutic versus subtherapeutic nasal continuous positive airway pressure on simulated steering performance in patients with obstructive sleep apnoea. AB - BACKGROUND: Obstructive sleep apnoea (OSA) impairs vigilance and may lead to an increased rate of driving accidents. In uncontrolled studies accident rates and simulated steering performance improve following treatment with nasal continuous positive airway pressure (NCPAP). This study seeks to confirm the improvement in steering performance in a randomised controlled trial using subtherapeutic NCPAP as a control treatment. METHODS: Fifty nine men with OSA (Epworth Sleepiness Score (ESS) of > or =10, and > or =10/h dips in SaO(2) of >4% due to OSA) received therapeutic or subtherapeutic NCPAP ( approximately 1 cm H(2)O) for one month. Simulated steering performance over three 30-minute "drives" was quantified as: standard deviation (SD) of road position, deterioration in SD across the drive, length of drive before "crashing", and number of off-road events. The reaction times to peripheral target stimuli during the drive were also measured. RESULTS: Subtherapeutic NCPAP did not improve overnight >4% SaO(2) dips/h compared with baseline values, thus acting as a control. The SD of the steering position improved from 0.36 to 0.21 on therapeutic NCPAP, and from 0.35 to 0.30 on subtherapeutic NCPAP (p = 0.03). Deterioration in SD of the steering position improved from 0.18 to 0.06 SD/h with therapeutic NCPAP and worsened from 0.18 to 0.24 with subtherapeutic NCPAP (p = 0.04). The reaction time to target stimuli was quicker after therapeutic than after subtherapeutic NCPAP (2.3 versus 2.7 seconds, p = 0.04). CONCLUSIONS: Therapeutic NCPAP improves steering performance and reaction time to target stimuli in patients with OSA, lending further support to the hypothesis that OSA impairs driving, increases driving accident rates, and that these improve following treatment with NCPAP. PMID- 10679543 TI - Non-invasive markers of airway inflammation as predictors of oral steroid responsiveness in asthma. AB - BACKGROUND: Sputum eosinophil counts and exhaled nitric oxide (NO) levels are increased in asthma and both measurements fall in response to corticosteroids. METHODS: Exhaled NO levels and sputum eosinophil counts were assessed as non invasive markers of the response to an oral steroid in 37 patients (19 women) with stable chronic asthma (mean (SD) age 48.6 (12.2) years, asthma duration 25. 9 (17.3) years, and baseline forced expiratory volume in one second (FEV(1)) 76.3 (21.9)% predicted). Spirometric tests, with reversibility to a beta agonist (2.5 mg nebulised salbutamol), and induced sputum (using nebulised 3% saline) were performed at recruitment and following treatment with 30 mg prednisolone/day for 14 days. RESULTS: Baseline NO levels correlated with the percentage improvement in FEV(1) from baseline to the post-steroid, post-bronchodilator value (r(s) = 0.47, p = 0.003), with an NO level of >10 ppb at baseline having a positive predictive value of 83% for an improvement in FEV(1) of > or =15% (sensitivity 59%, specificity 90%). Sputum eosinophilia (> or =4%) had a positive predictive value of 68% (sensitivity 54%, specificity 76%) for an increase in FEV(1) of > or =15%. A combination of sputum eosinophilia and increased NO levels resulted in a positive predictive value of 72% and a negative predictive value of 79% (sensitivity 76%, specificity 75%). CONCLUSION: Exhaled NO levels and sputum eosinophilia may be useful in predicting the response to a trial of oral steroid in asthma. PMID- 10679544 TI - Association of sputum parameters with clinical and functional measurements in asthma. PMID- 10679546 TI - Superior vena cava obstruction caused by radiation induced venous fibrosis. AB - Superior vena cava syndrome is most often caused by lung carcinoma. Two cases are described in whom venous obstruction in the superior mediastinum was caused by local vascular fibrosis due to radiotherapy five and seven years earlier. The development of radiation injury to greater vessels is discussed, together with the possibilities for treatment of superior vena cava syndrome. PMID- 10679545 TI - Asthma and poverty. PMID- 10679547 TI - Mitral stenosis obscuring the diagnosis of plexogenic pulmonary arteriopathy and familial pulmonary hypertension. AB - A patient who died after surgery for critical mitral stenosis was found to have underlying unrecognised plexogenic pulmonary arteriopathy and familial pulmonary hypertension. The importance of recognising familial pulmonary hypertension is discussed, together with the contribution of genetic and other risk factors to plexogenic pulmonary arteriopathy. PMID- 10679549 TI - Case Presentations in Clinical Tuberculosis. PMID- 10679548 TI - Bronchiolitis obliterans organising pneumonia associated with the use of nitrofurantoin. AB - The spectrum of nitrofurantoin lung injury continues to widen. The case histories are presented of two patients who developed lung disease associated with the use of nitrofurantoin with histological features of bronchiolitis obliterans organising pneumonia (BOOP), a rare but recognised form of drug induced injury. The two middle aged women presented with respiratory symptoms after prolonged treatment with nitrofurantoin. Both had impaired lung function and abnormal computed tomographic scans, and their condition improved when nitrofurantoin was withdrawn and corticosteroid treatment commenced. The favourable outcome in these two patients contrasts with the fatal outcome of the two other reported cases of nitrofurantoin induced BOOP. We suggest that the previous classification of nitrofurantoin induced lung injury into "acute" and "chronic" injury is an oversimplification in view of the wide variety of pathological entities that have subsequently emerged. PMID- 10679550 TI - Anti-Inflammatory Drugs in Asthma. PMID- 10679575 TI - Thrombin-induced phosphorylation of MARCKS does not alter its interactions with calmodulin or actin. AB - Myristoylated alanine-rich C kinase substrate (MARCKS) is a calmodulin (CaM)- and actin-binding protein and prominent protein kinase C (PKC) substrate. In vitro phosphorylation of MARCKS by PKC has been shown to induce the release of both CaM and actin, leading to the suggestion that MARCKS may regulate CaM availability during agonist-induced signalling. In support of this hypothesis we previously demonstrated that thrombin-induced MARCKS phosphorylation in endothelial cells (EC) parallels activation of myosin light chain kinase, a CaM-dependent enzyme. To test this theory further, we transfected CHO cells, which normally do not express significant levels of MARCKS, with a MARCKS cDNA. The thrombin-stimulated phosphorylation of myosin light chains and the sensitivity to CaM antagonists in the MARCKS overexpressing cells was the same as that in control CHO cells. MARCKS associated with the actin cytoskeleton in EC was markedly increased upon treatment with the PKC activator, PMA, but only modestly enhanced by thrombin treatment. Similarly, colocalisation of MARCKS with actin was enhanced when the EC were challenged with PMA but not thrombin. These data may be partially explained by PKC-independent phosphorylation of MARCKS in response to thrombin stimulation. PMID- 10679576 TI - Sphingolipid signalling domains floating on rafts or buried in caves? AB - Ceramide is a novel lipid mediator involved in regulating cell growth, cell differentiation and cell death. Many studies have focused on characterizing the stimulus-induced production of ceramide and identifying putative downstream molecular targets. However, little remains known about the localization of the regulated production of ceramide through sphingomyelin metabolism in the plasma membrane. Additionally, it is unclear whether a localized increase in ceramide concentration is necessary to facilitate downstream signalling events initiated by this lipid. Recent studies have suggested that detergent-insoluble plasma membrane domains may be highly localized sites for initiating signal transduction cascades by both tyrosine kinase and sphingolipid signalling pathways. These domains are typically enriched in both sphingolipids and cholesterol and have been proposed to form highly ordered lipid rafts floating in a sea of glycerophospholipids. Alternatively, upon integration of the cholesterol binding protein caveolin, these domains may also form small cave-like structures called caveolae. Emerging evidence suggests that the enhanced sphingomyelin content of these lipid domains make them potential substrate pools for sphingomyelinases to produce a high local concentration of ceramide. The subsequent formation of ceramide microdomains in the plasma membrane may be a critical factor in regulating downstream signalling through this lipid messenger. PMID- 10679577 TI - The essential role of H2O2 in the regulation of intracellular Ca2+ by epidermal growth factor in rat-2 fibroblasts. AB - We have investigated a new mechanism by which epidermal growth factor (EGF) increases intracellular Ca(2+) ([Ca(2+)](i)) in Rat-2 fibroblasts. EGF induced a transient increase of [Ca(2+)](i), and sustained Ca(2+) increase disappeared in the absence of extracellular Ca(2+). However, EGF had no effect on the formation of inositol phosphates. Expression of N17Rac or scrape-loading of C3 transferase blocked the elevation of [Ca(2+)](i) by EGF, but not by lysophosphatidic acid (LPA). EGF increased intracellular H(2)O(2), with a maximal increase at 5 min, which was blocked by catalase, scrape-loading of C3 transferase, or expression of N17Rac. H(2)O(2) scavengers, catalase and N-acetyl-L-cysteine, also blocked the Ca(2+) response to EGF, but not to LPA. In the presence of EGTA, preincubation with EGF completely inhibited subsequent Ca(2+) response to extracellular H(2)O(2) and vice versa. Incubation with EGF or phosphatidic acid abolished subsequent elevation of [Ca(2+)](i) by phosphatidic acid or EGF, respectively. Furthermore, preincubation with LPA inhibited the subsequent Ca(2+) response to EGF, but not vice versa. These results suggested that intracellular H(2)O(2) regulated by Rac and RhoA, but not inositol phosphates, was responsible for the EGF-stimulated elevation of [Ca(2+)](i). It was also suggested that EGF cross talked with LPA in the regulation of [Ca(2+)](i) by producing intracellular H(2)O(2). PMID- 10679578 TI - Potentiation of calcium levels by extracellular arachidonic acid in nuclei isolated from macrophages stimulated with receptor-recognized forms of alpha(2) macroglobulin. AB - Ligation of macrophage alpha(2)-macroglobulin signalling receptors (alpha(2)MSR) with activated alpha(2)-macroglobulin (alpha(2)M*) increases intracellular Ca(2+), and cytosolic phospholipase A(2) (cPLA(2)) and phospholipase D activities. In view of the relationship between cellular Ca(2+) and mitogenesis, we examined the effect of the product of cPLA(2) activity, arachidonic acid (AA), on nuclear Ca(2+) levels in macrophages stimulated with alpha(2)M*, platelet derived growth factor, and bradykinin. AA addition increased Ca(2+) levels in Fura-2/AM loaded nuclei from both buffer-treated and agonist-stimulated cells, but the increase in stimulated macrophages was 2-4-fold higher. Preincubation of Fura-2/AM loaded nuclei with EGTA or BAPTA/AM abolished AA-induced increase in nuclear Ca(2+) levels. Preincubation of nuclei with indomethacin did not affect AA-induced increase in nuclear Ca(2+) in agonist-stimulated nuclei. It is concluded that in macrophages stimulated with various agonists, AA, derived from cPLA(2)-dependent hydrolysis of phospholipids, plays a significant role in regulating nuclear Ca(2+) levels and thus nuclear functions. PMID- 10679579 TI - Protein kinase and phosphatase modulation of quail brain GABA(A) and non-NMDA receptors co-expressed in Xenopus oocytes. AB - The GABA(A) receptor and the non-NMDA subtype of the ionotropic glutamate receptor were co-expressed in Xenopus oocytes by injection of quail brain mRNA. The oocytes were treated with various protein kinase (PK) and protein phosphatase (PP) activators and inhibitors and the effects on receptor functioning were monitored. Two phorbol esters, 4-beta-phorbol 12-myristate-13-acetate (PMA) and 4 beta-phorbol 12,13-dibutyrate (PDBu); the cGMP-dependent PK activators sodium nitroprusside (SNP) and S-nitrosoglutathione (SNOG); and the PP inhibitor okadaic acid (OA) reduced the amplitude of the GABA-induced currents, whilst the PK inhibitor staurosporine potentiated it. In addition, PMA, PDBu, SNP, and OA reduced the desensitization of the GABA-induced response. Identical treatments generally had similar but less pronounced effects on responses generated by kainate (KA) but the desensitization characteristic of the non-NMDA receptor was not affected. None of the treatments had any effect on the reversal potentials of the induced currents. Immunoblots revealed that the oocytes express endogenous PKG and guanylate cyclase. The results are discussed in terms of the molecular structures of GABA(A) and non-NMDA receptors and the potential functional consequences of phosphorylation/dephosphorylation. PMID- 10679580 TI - Evidence for stimulation of adenylyl cyclase by an activated G(s) heterotrimer in cell membranes: an experimental method for controlling the G(s) subunit composition of cell membranes. AB - Heterotrimeric (alphabetagamma) G(s) mediates agonist-induced stimulation of adenylyl cyclase (AC). Cholera toxin (CTx) will ADP-ribosylate the alpha-subunit of G(s) (G(s)alpha). G(s)alpha-deficient cyc(-) membranes were "stripped" of Gbeta. When the stripped cyc(-) were incubated with G(s)alpha and/or Gbetagamma, each was incorporated into the membranes independently of the other. Both G(s)alpha and Gbetagamma had to be present in the membranes, and they had to be able to form a heterotrimer in order for CTx to ADP-ribosylate G(s)alpha, indicating that the membrane bound G(s) heterotrimer is a substrate for CTx, but the G(s)alpha subunit by itself is not. When G(s)alpha was completely and irreversibly activated with GTPgammaS and incorporated into stripped cyc(-), it was a poor substrate for CTx and a weak stimulator of AC unless Gbetagamma was also incorporated. Furthermore, the level of AC stimulation corresponded to the amount of G(s) heterotrimer that was formed in the membranes from GTPgammaS activated G(s)alpha and Gbetagamma. These data suggest that AC is stimulated by an activated G(s) heterotrimer in cell membranes. PMID- 10679581 TI - [The journal enters the new millennium. A readership survey: why?]. PMID- 10679582 TI - [Intraductal mucinous papillary tumors of the pancreas: which procedure for which tumor?]. PMID- 10679583 TI - [Megaduodenum in chronic intestinal pseudo-obstruction: management by duodenectomy-duodenoplasty]. AB - BACKGROUND: Surgical management of primitive chronic intestinal pseudo obstruction involving the duodenum (megaduodenum) is an uncommon but still difficult problem. PATIENTS AND METHODS: Six patients who experienced severe symptoms were managed by an original surgical procedure including partial duodenal resection and reconstruction of a duodenal tract using a large duodenal anastomosis (duodenectomy-duodenoplasty). RESULTS: There was no postoperative complication. All preoperative symptoms completely regressed in all but one patient who had previously undergone a vagotomy and experienced transient early post-operative gastric stasis. With a median follow-up of 6 years (range 4-9), all patients had good functional results without any evidence of other motility disorders. The mean weight gain was 10 kg (range 7-15). CONCLUSIONS: Duodenectomy duodenoplasty is a safe procedure resulting in efficient symptom relief in patients suffering from megaduodenum. PMID- 10679584 TI - [Intraperitoneal cisplatin plus epinephrine and surgical debulking for the treatment of advanced peritoneal carcinomatosis in the rat]. AB - OBJECTIVE: To evaluate and compare the effects of cytoreductive surgery with intraperitoneal cisplatin and epinephrine on peritoneal carcinomatosis in the rat. MATERIAL AND METHODS: Twenty-day old peritoneal carcinomatosis was obtained after intraperitoneal injection of 1 x 10(6) DHD/K12/PROb cells into BDIX rats. The surgical treatment included electric fulguration of the peritoneal tumors with spleen and omentum removal while intraperitoneal chemotherapy included platinum (3 mg/kg) associated with epinephrine (2 mg/kg). RESULTS: Surgery did not increase rat survival unlike cisplatin or cisplatin/epinephrine chemotherapy. Intraperitoneal chemotherapy alone with cisplatin +/- epinephrine increased survival but did not provide cure. Surgery followed by intraperitoneal cisplatin and epinephrine cured four out of five twenty-day old peritoneal carcinomatosis. CONCLUSION: Surgery combined with intraperitoneal cisplatin and epinephrine could be an efficient treatment for peritoneal carcinomatosis in man. PMID- 10679585 TI - Low sensitivity of invasive tests for the detection of Helicobacter pylori infection in patients with bleeding ulcer. AB - BACKGROUND: A high false negative rate for antral infection with Helicobacter pylori when assessed by rapid urease test has recently been reported in patients with bleeding ulcer. This result could partly explain the differing prevalence of H. pylori infection in bleeding and non-bleeding ulcers. AIMS: To evaluate the accuracy of a rapid urease test (UT), histology and culture for detection of H. pylori in antral biopsies from acute bleeding peptic ulcer patients using a serological test as reference. PATIENTS AND METHODS: All consecutive patients with active bleeding gastric or duodenal ulcer at endoscopic examination admitted in six university hospitals in France were considered for inclusion. Five antral biopsies were taken during the diagnostic endoscopy for UT, culture and histology. A blood sample was taken for H. pylori serology. RESULTS: One hundred and eighty one patients were included and 129 (71%) had a positive serology. The sensitivity of UT, histology and culture for detection of H. pylori infection were 41%, 33% and 34%, respectively. The sensitivity and specificity of the combination of the three invasive tests were 48.8% (95% CI: 40.2-57.4) and 90.6% (95% CI: 82. 6-99) respectively. In the 52 serologically negative patients, only 5 had at least one invasive positive test. The sensitivity of the invasive tests decreased significantly with age but was not influenced by NSAIDs intake. Of 80 patients with a positive serological test and negative histological evaluation for H. pylori, chronic antral inflammation was found in 70 patients (87%). In 46 patients with both negative serological test and H. pylori negative test according to histology, only 13 (28%) had chronic antral inflammation. CONCLUSIONS: The sensitivity of invasive tests for detection of H. pylori is low during acute ulcer bleeding, and they should be used with caution in this condition. A serological test is recommended to identify patients with H. pylori infection in spite of negative invasive tests. PMID- 10679586 TI - [Anoperineal manifestations of Crohn's disease]. PMID- 10679587 TI - [Clinical hepatology at the dawn of the year 2000]. PMID- 10679588 TI - [Prevalence of hepatitis C in patients with chronic inflammatory bowel disease in the region of Nice and evaluation of risk factors]. AB - OBJECTIVES: To estimate the prevalence of viral hepatitis C markers and to determine independent risk factors in a population of patients with inflammatory bowel disease. METHODS: We studied 117 consecutive out-patients (male/female, 53/64; mean age 41 +/- 16 yrs) with ulcerative colitis (43 patients) or Crohn's disease (74 patients). Anti-hepatitis C virus antibodies were tested with a third generation Elisa test. The following risk factors were tested for each patient: duration of inflammatory bowel disease, number of colonoscopies, history of surgical procedures, blood transfusions, intravenous drug abuse and immunosuppressive treatments. RESULTS: The seroprevalence of hepatitis C virus was 5.98% (7/117). The only risk factor independently associated with serological markers for hepatitis C virus was blood transfusion (odds ratio: 7.77; confidence interval: 95% (1.63-49.09); P=0.012). CONCLUSIONS: The prevalence of hepatitis C virus infection was high in patients with inflammatory bowel disease, mainly due to blood transfusions. Colonoscopies and surgical procedures were not found to be additional risk factors for infection with hepatitis C virus. PMID- 10679589 TI - [The Hepatitis Network in Maine-et-Loire city hospitals: six year follow-up of patients and physicians. Hepatitis Network of Maine-et-Loire]. AB - OBJECTIVES: To evaluate 6 years of a city-hospital hepatitis network. The network was set up in 3 steps: 1988: intrahospital network, 1991: city-hospital network, 1997: compliance with government regulations. METHODS: The whole activity from 1991 to 1997 was evaluated and special attention was paid to patient files and participating physicians. RESULTS: From June 1991 to December 1997 (6.5 years), 759 patient files were registered which corresponds to 531 patients (male 57%) with a mean age of 44 +/- 16 years (+/- standard deviation). Four hundred and twenty one patients (79%) had hepatitis C, 95 (18%) hepatitis B and 15 (3%) co infection; 83% of patients had had a liver biopsy confirming cirrhosis in 21.5%. The annual number of files registered increased continuously. This was more a result of recruiting known patients than new patients, after the network had been in place for several years, mainly with hepatitis B virus (known patients in 1997: hepatitis B virus: 53% vs 33% for hepatitis C virus, P<0.05). Treatment protocols (73%) were more frequent for hepatitis C virus patients than for hepatitis B (73% vs 59%, P<0.01). Therapeutic trial proposals (37%) increased from 21% in 1991 to 59% in 1997, P<0.01. Participation in monthly meetings by academic hepato-gastroenterologists increased slightly while that of regional hospital hepato-gastroenterologists increased markedly and that of private hepato gastroenterologists remained stable. The annual proportion of files submitted by academic hepato-gastroenterologists decreased in parallel to the increase in submission of patient files by other hepato-gastroenterologists. CONCLUSIONS: During 6 years of activity, the network grew with an increase in the annual number of patient files, growing participation in therapeutic trials as well as in monthly meetings by practitioners. PMID- 10679590 TI - [Hepatic vascular malformations in Rendu-Osler disease]. PMID- 10679591 TI - [Pre-test]. PMID- 10679592 TI - [Treatment of gastrointestinal bleeding in patients with cirrhosis]. PMID- 10679593 TI - [Portal hypertension: primary prevention]. PMID- 10679594 TI - [Hemorrhage caused by ruptured esophageal varices in acute alcoholic hepatitis in a patient with cirrhosis]. PMID- 10679595 TI - [Questions to Pr. P. Cales]. PMID- 10679596 TI - [Answers to the pre-test]. PMID- 10679597 TI - [Mesenteric localization of Castleman's disease]. AB - Castleman's disease (or angiofollicular lymph node hyperplasia) is a rare pathologic process of unknown etiology. Commonly, it is an unique tumor localized in the mediastinum (unicentric disease). Extrathoracic localizations are however reported with an increasing frequency. We describe two cases of mesenteric Castleman's disease. Diagnosis can be suggested by preoperative morphologic explorations (ultrasonography with Doppler, computed tomography scan, magnetic resonance imaging and angiography) and confrontation with clinical and biological data (young age, biological signs of inflammation). In the classical unique tumor of hyalin-vascular type, surgical removal allows definitive cure. Prognosis is poor in the plasma cell type, mostly in multicentric disease due to a high incidence of malignancy. PMID- 10679598 TI - [Enucleation of intraductal papillary-mucinous tumor of the head of the pancreas. Report of 2 cases]. AB - Intraductal papillary-mucinous tumors of the pancreas are characterized by malignant transformation of unpredictable occurrence because of their unknown natural history. Surgical treatment is duodenopancreatectomy or left pancreatectomy even for benign tumors. We report 2 cases of benign intraductal papillary-mucinous tumor confined to the head of the pancreas and treated by enucleation. PMID- 10679599 TI - [Emergence of resistant hepatitis B virus strains during long-term lamivudine therapy in human immunodeficiency virus co-infected patients]. AB - Seven patients co-infected with hepatitis B virus (HBsAg and HBeAg carriers, quantifiable HBV DNA with the bDNA technic) and human immunodeficiency virus received a triple antiretroviral combination therapy, including lamivudine (150 mg twice a day). Hepatitis B viral load rapidly became undetectable in 6/7 patients. It remained below the level of detection in 2 subjects, after 20 and 22 months of treatment, with one of them achieving HBeAg/anti-HBe seroconversion. However, in the other 4 individuals, hepatitis B viremia increased again after 8 to 16 months of lamivudine-containing regimen. The last patient was a non responder. The 4 relapsers developed a double mutation Leu(528) for Met(528) and Met(552) for Val(552), on hepatitis B virus polymerase, either concomitant (M8 and M16) with a hepatitis B virus DNA increase, or 2 months earlier (M10 and M12). The high frequency of hepatitis B virus resistance to lamivudine emphasizes the necessity of identifying more effective strategies, such as double combination therapies. PMID- 10679600 TI - [Hemorrhagic rectocolitis following liver transplantation for alcoholic cirrhosis]. PMID- 10679601 TI - Geriatric nursing comes of age. PMID- 10679602 TI - Research review committees in long-term care settings. AB - As the number of elderly people grows, the interest in research for this population increases. Federal funding is available for research regarding elders' needs, and researchers are recognizing the problems this population incurs. Residents and staff members in long-term care facilities are prime candidates for study subjects, and institutional research review committees, supported by the facilities' administration, are necessary to protect them. This article discusses ways to establish a research review committee and its working processes. PMID- 10679603 TI - A family perspective of family/staff interaction in long-term care facilities. AB - This article explores the ways that families perceive their relationships with the staff in long-term care facilities. Qualitative interviews were conducted with 27 families who had elder members in two facilities in Ontario, Canada. The families identified the staff characteristics that they valued and the ways they developed relationships with staff members. PMID- 10679604 TI - Comparison of pain assessment instruments in cognitively intact and cognitively impaired nursing home residents. AB - This study was conducted to determine which pain severity and location instruments were most useful in the nursing home setting. Pain severity and location were assessed monthly for 1 year in 37 participants enrolled in a restorative rehabilitation program. Pain location was determined by the residents' indications on a diagram, a doll, and their body. Pain severity was determined by resident response to verbal, visual analog, faces, and word scales. Cognitively impaired residents had greater difficulty using all instruments. The McGill Word Scale was used most to determine pain severity. Pointing to themselves most frequently determined pain location among residents. New strategies are needed for pain assessment in the elderly, especially the cognitively impaired elderly, and a combination of instruments to assess pain in the latter group may be necessary. PMID- 10679605 TI - Joyce Springate, EdD, RN,Contiguous Canadian Sister and Colleague. PMID- 10679606 TI - Proudly professional: the Canadian Gerontological Nursing Association. PMID- 10679607 TI - Aging and meaning in life: examining the concept. AB - To make sense of their existence in the face of adversity and chaos or during times of relative calm, human beings seek meaning. Meaning has been identified as a significant factor in health and well-being in later years. Engaging in an intensive literature search about meaning in later life constituted the beginning step for a qualitative research study. By studying how older people experience meaning in their lives, we may learn more about the human experiences of joy and hope and their capacity to respond to opportunities and manage problems in their lives. PMID- 10679608 TI - Specialized care unit: family and staff perceptions of significant elements. AB - Specialized care units (SCUs) for nursing home residents with dementia are increasingly prevalent. Although reported to promote positive resident outcomes, the specific characteristics of SCUs most likely to yield these outcomes have not yet been identified. This article presents the findings of a qualitative study that investigated the perceptions of family members and staff about SCU characteristics that contribute to positive outcomes for residents with dementia. All family and staff members interviewed agreed that the SCU fostered feelings of personal space, personhood, and an unforced routine. These findings are discussed and provide specific direction for care of nursing home residents with dementia. PMID- 10679609 TI - Life albums in long-term care: resident, family, and staff perceptions. AB - A life album is a collection of selected memorabilia, photographs, and archival and other material that describes an individual's life in a photograph book format. During a summer research program, 13 long-term care residents created life albums that contained material from their past and current life. Some pages were left unfilled to allow material to be added in the future. These albums provided residents, families, and staff members with a means to recall life events. This article reports their perceptions of creating such albums. PMID- 10679610 TI - Resident abuse: an insider's perspective. AB - The purpose of this study was to assess how resident abuse is perceived by members of the long-term care institution. Groups of registered nurses, nonprofessional staff, older residents, and significant others were interviewed individually and participated in focus groups. Resident abuse is perceived by the participants in this study as an experience that causes a perception of hurt in older residents. This perception is voiced by either older residents themselves or other members of the institution on their behalf. Participants' views about resident abuse are always framed within the context of institutional life. PMID- 10679611 TI - GN management leaders: Cheryl Ciechomski and Bernardine Gorek. Interview by Priscilla Ebersole. PMID- 10679612 TI - Jog your memory: a community screening model. AB - This study's objectives were to establish an annual memory-screening week, heighten public awareness of the issues surrounding cognitive impairments, promote baseline cognitive screening as part of routine examinations in primary care settings for the older adult, and detect early dementia and depression. The 264 participants completed demographic and anecdotal information and the Geriatric Depression Scale (GDS). Nurses scored the GDS and administered the Mini Mental State Examination (MMSE) and the Clock Drawing Test. Screening results and educational information were given to the participants. Results revealed that 14% of participants scored abnormally on the MMSE and approximately 14% had an abnormal GDS score. PMID- 10679613 TI - Advising older adults about pain remedies. PMID- 10679614 TI - Telecaring in home care: making a telephone visit. PMID- 10679616 TI - High-frequency dielectric study of side-chain dynamics in poly(lysine) aqueous solutions. AB - The high-frequency dielectric properties of poly(lysine) of different chirality in aqueous solutions have been measured in the frequency range from 1 MHz to 1.8 GHz. The dielectric spectra show the existence of relatively small dielectric dispersions at around 100 MHz that have been attributed to internal motion in the polymer chain, due to side-chain polar groups. Our results indicate that the local structure of the chain and its possibility to undergo a conformational transition induced by pH does not modify the main feature of the side-chain dynamics, the dielectric strength being largely proportional to the concentration of charged groups. A similar behavior has been found in poly(alpha-glutamate) and in poly(gamma-glutamate) aqueous solutions, where the dielectric parameters appear to be related to the change in the charge density on the main chain, rather than to the accompanying conformational helix-coil transition. PMID- 10679615 TI - The third-dimensional structure of the complex between an Fv antibody fragment and an analogue of the main immunogenic region of the acetylcholine receptor: a combined two-dimensional NMR, homology, and molecular modeling approach. AB - Binding of autoantibodies to the acetylcholine receptor (AChR) plays a major role in the autoimmune disease Myasthenia gravis (MG). In this paper, we propose a structure model of a putative immunocomplex that gives rise to the reduction of functional AChR molecules during the course of MG. The model complex consists of the [G(70), Nle(76)] decapeptide analogue of the main immunogenic region (MIR), representing the major antigenic epitope of AChR, and the single chain Fv fragment of monoclonal antibody 198, a potent MG autoantibody. The structure of the complexed decapeptide antigen [G(70), Nle(76)]MIR was determined using two dimensional nmr, whereas the antibody structure was derived by means of homology modeling. The final complex was constructed using calculational docking and molecular dynamics. We termed this approach "directed modeling," since the known peptide structure directs the prestructured antibody binding site to its final conformation. The independently derived structures of the peptide antigen and antibody binding site already showed a high degree of surface complementarity after the initial docking calculation, during which the peptide was conformationally restrained. The docking routine was a soft algorithm, applying a combination of Monte Carlo simulation and energy minimization. The observed shape complementarity in the docking process suggested that the structure assessments already led to anti-idiotypic conformations of peptide antigen and antibody fragment. Refinement of the complex by dynamic simulation yielded improved surface adaptation by small rearrangements within antibody and antigen. The complex presented herein was analyzed in terms of antibody-antigen interactions, properties of contacting surfaces, and segmental mobility. The structural requirements for AChR complexation by autoantibodies were explored and compared with experimental data from alanine scans of the MIR peptides. The analysis revealed that the N-terminal loop of the peptide structure, which is indispensable for antibody recognition, aligns three hydrophobic groups in a favorable arrangement leading to the burial of 40% of the peptide surface in the binding cleft upon complexation. These data should be valuable in the rational design of an Fv mutant with much improved affinity for the MIR and AChR to be used in therapeutic approaches in MG. PMID- 10679617 TI - Arg side-chain-backbone interactions evidenced in model peptides by 17O-NMR spectroscopy. AB - The guanidinium group of arginine possesses a variety of biochemical functions, either by participating in direct interactions in recognition processes, or by stabilizing secondary structures. Three model compounds, selectively (17)O enriched, Ac-Arg-Ala-[(17)O]Pro-NH(2) (1), Piv-Arg-Pro-[(17)O]Gly-NH(2) (2) (C terminal segment of the luteinizing hormone releasing hormone), and Piv-Nle-Pro [(17)O]Gly-NH(2) (3), were prepared and studied by (17)O-nmr spectroscopy. A direct hydrogen-bonded interaction between the Arg side chain and the carbonyl main chain carboxy-terminus was found, thus confirming the tendency of Arg to participate in proton-acceptor functions. PMID- 10679618 TI - Effect of lengthening of peptide backbone by insertion of chiral beta-homo amino acid residues: conformational behavior of linear peptides containing alternating L-leucine and beta-homo L-leucine residues. AB - The synthesis and the solution behavior of the linear peptides containing a beta homo (beta-H) leucine residue-Boc-Leu-beta-HLeu-Leu-OMe, Boc-beta-HLeu-Leu-beta HLeu-Leu-OMe, and Boc-Leu-beta-HLeu-Leu-beta-HLeu-Leu-OMe-as well as the solid structure of the tripeptide, are reported. The conformational behavior of the peptides was investigated in solution by two-dimensional nmr. Our data support the existence in solution with different families of conformers in rapid interchange. The crystals of the tripeptide are orthorhombic, space group P2(1)2(1)2, with a = 15.829(1) A, b = 29.659(1) A, c = 6.563(1) A, and Z = 4. The structure has been solved by direct methods and refined to final R1 and wR2 indexes of 0.0530 and 0.1436 for 2420 reflections with I > 2sigma(I). In the solid state, the tripeptide does not present intramolecular H bonds, and the peptide backbone of the two leucine residues adopts a quasi-extended conformation. For the beta-HLeu residue, the backbone conformation is specified by the torsion angles straight phi(2) = -120.9(4) degrees, mu(2) = 56.7(4) degrees, psi(3) = -133.2(4) degrees. The side chains of the three residues assume the same conformation (g(-), g(-), trans), and all peptide bonds, except the urethane group at the N-terminus, are in the trans conformation. Preliminary conformational energy calculations carried out on the Ac-NH-beta-HAla-NHMe underline that the conformations with mu angle equal to 180 degrees and 60 degrees assume lower energy with respect to the others. In addition, we found a larger conformational freedom for the psi angle with respect to the straight phi angle. PMID- 10679619 TI - The crystal structure of Afc-containing peptides. AB - A systematic structural analysis of Afc (9-amino-fluorene-9-carboxylic acid) containing peptides is here reported. The crystal structures of four fully protected tripeptides containing the Afc residue in position 2: Z-X(1)-Afc(2) Y(3)-OMe (peptide a: X = Y = Gly; peptide b: X = Aib, C(alpha, alpha) dimethylglycine, Y = Gly; peptide c: X = Gly, Y = Aib; peptide d: X = Y = Aib) have been solved by x-ray crystallography. All the results suggest that the Afc residue has a high propensity to assume an extended conformation. In fact, the Afc residue adopts an extended conformation in three peptides examined in this paper (peptides a-c). In contrast, Afc was found in a folded conformation, in the 3(10)-helical region, only in the peptide d, in which it is both preceded and followed by the strong helix promoting Aib. PMID- 10679620 TI - Conformational behavior of C alpha,alpha-diphenyl glycine: extended conformation in tripeptides containing consecutive D phi G residues. AB - Recent studies on the conformational preferences of the Dphig (C(alpha,alpha) diphenylglycine) residue showed that this C(alpha,alpha)-disubstituted glycine has a structural versatility. In fact, depending on the nature of the following or preceding residue, Dphig can assume either folded or extended conformations. We have carried out the analysis of the conformational preferences of the Dphig residue in tripeptides containing consecutive Dphig residues. The crystal structures of Z-Dphig-Dphig -Dphig-OMe (a; Z = benzyloxycarbonyl; OMe = methyl ester), Z-Aib-Dphig-Dphig-OMe (b; Aib = alpha-aminoisobutyric acid), and Z-Ac(3)c Dphig-Dphig-OMe (c; Ac(3)c = alpha-amino-cyclopropan carboxylic acid), are here reported. The Dphig residues adopt the fully extended conformation in the three tripeptides examined. Together with our previous findings on Dphig containing peptides, the structures of the peptides here examined, indicate that the presence of adjacent Dphig residue in the sequence further stabilizes the extended conformation. PMID- 10679621 TI - A spectroscopic and molecular mechanics investigation on a series of AIB-based linear peptides and a peptide template, both containing tryptophan and a nitroxide derivative as probes. AB - Linear Aib-based hexapeptides, of the general formula Ac-Toac-(Aib)(n) -Trp (Aib)(r) -OtBu [T(Aib)(n) Trp], where n + r = 4, and Toac is a nitroxide spin labeled C(alpha,alpha)-disubstituted glycine, were investigated by steady-state and time-resolved fluorescence measurements in different solvent media. A related peptide, i.e., cyclo-?Orn-[(Aib)(2)-Trp-(Aib)(2)-Z]-Asp-[(Aib)(2)-Toac-(Aib)(2)-+ ++OtBu ]? [T-cyclo-Trp], was also studied by the same techniques. It is a L-Orn, L-Asp diketopiperazine template, to which two Aib-based chains are covalently attached, each one containing one chromophore only, i.e., Trp or Toac. Whatever the solvent, in the former series of peptides quenching of the excited Trp exhibits three lifetime components and proceeds on a time scale from subnanoseconds to a few nanoseconds, while in the case of the template the same process occurs entirely on the nanoscale time scale, exhibiting two lifetimes only. The ir absorption spectral patterns suggest that the backbone of the peptides examined is in the 3(10)-helical conformation, as earlier determined by x-ray diffraction for T(Aib)(3)Trp in the crystal state. In all cases, the fluorescence results are satisfactorily described by a dipole-dipole interaction mechanism, in which electronic energy transfer takes place from the excited Trp to Toac, provided the mutual orientation between the fluorophore and Toac is taken into account. This implies that interconversion among conformational substates is slow on the time scale of the transfer process, allowing us to estimate the dynamics of the process. Molecular mechanics calculations coupled with time decay data made it possible to build up the most probable structures of these peptides in solution. PMID- 10679622 TI - The crystal structure of a Dcp-containing peptide. AB - We have investigated the conformational preferences of a newly synthesized C(alpha,alpha) symmetrically disubstituted glycine, namely alpha,alpha dicyclopropylglycine (Dcp). We report here the crystal structure of a fully protected dipeptide containing Dcp, namely Z-Dcp(1)-Dcp(2)-OCH(3). Both Dcp residues are in a folded conformation. The overall peptide structural organization corresponds to an alpha-pleated sheet conformation, similar to that observed in linear peptides made up of alternating D- and L-residues and in Z-Aib Aib-OCH(3) (Aib: alpha,alpha-dimethylglycine). These preliminary data suggest that the Dcp could represent an alternative as molecular tool to stabilize folded conformations. PMID- 10679623 TI - Solid state structural analysis of the cyclooctapeptide cyclo- (Pro1-Pro-Phe-Phe Ac6c-Ile-D-Ala-Val8). AB - A solid state analysis of the cyclic octapeptide c(-Pro(1)-Pro-Phe-Phe-Ac(6)c-Ile D-Ala-Val(8)-) (C8-CLA), containing the Pro-Pro-Phe-Phe sequence, followed by the bulky helicogenic C(alpha,alpha)-dialkylated 1-aminocyclohexane-1-carboxylic acid (Ac(6)c) residue and a D-Ala residue in position 7, has been carried out by x-ray diffraction. The crystals, grown from a DMSO solution, are monoclinic, space group P2(1) with a = 13.458(3) A, b = 19. 404(5) A, c = 21.508(4) A, and beta = 90.83(6) degrees, with two independent cyclic molecules in the asymmetric unit, two DMSO molecules, and three water molecules. The structure has been solved using the half and bake procedure by Sheldrick, and refined to final R1 and wR2 indices of 0.0613 and 0.1534 for 9867 reflections with I > 2sigma(I). This cyclic peptide, a deletion analogue of the naturally occurring cyclic nonapeptide cyclolinopeptide A [c(Pro-Pro-Phe-Phe-Leu-Ile-Ile-Leu-Val), CLA] has been designed to study the influence of the ring size reduction on the conformational behavior of CLA and more in general to obtain structural information on asymmetric cyclic octapeptides. The compound exhibits, in the solid state, a "banana-twisted" conformation with a cis peptide bond located between the two proline residues. Five intramolecular H bonds stabilize the structure: one type VIa beta-turn, two consecutive type III/I beta-turns, one gamma-turn, and one C(16) bend. The structure has also been compared with either the solution structure previously reported by us and obtained by nmr and computational analysis, and with solid state structural data reported in the literature on cyclic octapeptides. PMID- 10679624 TI - Conformational restriction through C alpha i <--> C alpha i cyclization: Ac12c, the largest cycloaliphatic C alpha,alpha- disubstituted glycine known. AB - Two complete series of N-protected, monodispersed oligopeptide esters to the pentamer level from 1-aminocyclododecane-1-carboxylic acid (Ac(12)c), an alpha amino acid conformationally constrained through C(alpha)(i) <--> C(alpha)(i) cyclization, and either L-Ala or Aib residues, along with the N-protected Ac(12)c homopeptide alkylamide series from monomer to trimer, have been synthesized by solution methods and fully characterized. The solution-preferred conformations of these peptides have been assessed by Fourier transform ir absorption and (1)H-nmr techniques. Moreover, the molecular structures of one derivative (Z-Ac(12)c-OH) and three peptides [the tripeptide ester Z-L-Ala-Ac(12)c-L-Ala-OMe, the tripeptide alkylamide Z-(Ac(12)c)(3)-NHiPr, and the tetrapeptide ester Z-(Aib)(2) Ac(12)c-Aib-OtBu (Aib, alpha-aminoisobutyric acid)] have been determined in the crystal state by x-ray diffraction. The results obtained point to the conclusion that beta-bends and 3(10)-helices are preferentially adopted by peptides based on Ac(12)c, the largest cycloaliphatic C-disubstituted glycine known. A comparison with the structural tendencies extracted from published works on peptides from Aib, the prototype of C-disubstituted glycines, and the other extensively studied members of the class of 1-aminocycloalkane-1-carboxylic acids (Ac(n) c, with n = 3-9), is made and the implications for the use of the Ac(12)c residue in the Ac(n) c scan approach of conformationally restricted analogues of bioactive peptides are briefly discussed. PMID- 10679625 TI - Effect of lysine residues on the deamidation reaction of asparagine side chains. AB - The effect of lysine residues on the deamidation reaction of the asparagine side chain has been studied on the peptide and on its lysine-acetylated derivative in a wide range of pH values. The amino acid sequence of these peptides is similar to the local sequence flanking the labile Asn-67 in RNAse A. The experimental data show that Lys influences both the deamidation rate and the relative yield of the two reaction products, i.e., the aspartic acid and beta-aspartic acid containing peptide. These effects are pH dependent and can be rationalized based on the mechanism previously proposed for the deamidation reaction via succinimide derivative. PMID- 10679626 TI - How the protein concentration affects unfolding curves of oligomers. AB - The position of unfolding curves of oligomeric proteins depends on the protein concentration. The extent of this dependence is analyzed here in terms of the midpoint concentration, i.e., the denaturant concentration at which the fractions of folded and unfolded protein are equal. Reexamination of published data highlights that the midpoint concentration decreases as the protein concentration becomes lower, as expected. Moreover, there are differences between urea and guanidine hydrochloride, as well as discrepancies between the linear extrapolation model and the denaturant binding model. These discrepancies could be used to choose the denaturation model that best fits experimental data. The equations used can be applied to any oligomeric system to check the validity of the two-state assumption. PMID- 10679627 TI - Amino acid preferences of small, naturally occurring polypeptides. AB - An analysis of amino acid composition of small, naturally occurring peptides ranging in size from 3 to 50 residues has been carried out. The purpose of the study is to determine whether differential trends in amino acid usage exist for small peptides compared to larger polypeptides and proteins. Results indicate that Cys, Trp, and Phe are substantially more frequent in peptides compared to their abundance in proteins at large. Aliphatic hydrophobic residues, particularly Leu and Ile, are somewhat underrepresented, while the frequency of Glu is significantly reduced. The shorter peptides are also more frequently neutral and become increasingly charged as their size increases. PMID- 10679628 TI - Evidence for a strong sulfur-aromatic interaction derived from crystallographic data. AB - We have uncovered new evidence for a significant interaction between divalent sulfur atoms and aromatic rings. Our study involves a statistical analysis of interatomic distances and other geometric descriptors derived from entries in the Cambridge Crystallographic Database (F. H. Allen and O. Kennard, Chem. Design Auto. News, 1993, Vol. 8, pp. 1 and 31-37). A set of descriptors was defined sufficient in number and type so as to elucidate completely the preferred geometry of interaction between six-membered aromatic carbon rings and divalent sulfurs for all crystal structures of nonmetal-bearing organic compounds present in the database. In order to test statistical significance, analogous probability distributions for the interaction of the moiety X-CH(2)-X with aromatic rings were computed, and taken a priori to correspond to the null hypothesis of no significant interaction. Tests of significance were carried our pairwise between probability distributions of sulfur-aromatic interaction descriptors and their CH(2)-aromatic analogues using the Smirnov-Kolmogorov nonparametric test (W. W. Daniel, Applied Nonparametric Statistics, Houghton-Mifflin: Boston, New York, 1978, pp. 276-286), and in all cases significance at the 99% confidence level or better was observed. Local maxima of the probability distributions were used to define a preferred geometry of interaction between the divalent sulfur moiety and the aromatic ring. Molecular mechanics studies were performed in an effort to better understand the physical basis of the interaction. This study confirms observations based on statistics of interaction of amino acids in protein crystal structures (R. S. Morgan, C. E. Tatsch, R. H. Gushard, J. M. McAdon, and P. K. Warme, International Journal of Peptide Protein Research, 1978, Vol. 11, pp. 209 217; R. S. Morgan and J. M. McAdon, International Journal of Peptide Protein Research, 1980, Vol. 15, pp. 177-180; K. S. C. Reid, P. F. Lindley, and J. M. Thornton, FEBS Letters, 1985, Vol. 190, pp. 209-213), as well as studies involving molecular mechanics (G. Nemethy and H. A. Scheraga, Biochemistry and Biophysics Research Communications, 1981, Vol. 98, pp. 482-487) and quantum chemical calculations (B. V. Cheney, M. W. Schulz, and J. Cheney, Biochimica Biophysica Acta, 1989, Vol. 996, pp.116-124; J. Pranata, Bioorganic Chemistry, 1997, Vol. 25, pp. 213-219)-all of which point to the possible importance of the sulfur-aromatic interaction. However, the preferred geometry of the interaction, as determined from our analysis of the small-molecule crystal data, differs significantly from that found by other approaches. PMID- 10679629 TI - An examination of the steric effects of 5-tert-butylproline on the conformation of polyproline and the cooperative nature of type II to type I helical interconversion. AB - The influence of steric effects on the helical geometry and the interconversion of type II to type I polyproline in water was examined by the synthesis and analysis of proline dimers and hexamers containing up to three (2S,5R)-5-tert butylproline residues. In the dimers, the bulky 5-tert-butyl substituent was found to exert a significant influence on the local prolyl amide geometry such that the predominant trans-isomer in N-(acetyl)prolyl-prolinamide (1) was converted to 63% cis isomer in N-(acetyl)prolyl-5-tert-butylprolinamide (2) as measured by (1)H-nmr spectroscopy. Similarly, the presence of a 5-tert-butyl group on the C-terminal residue in the polyproline hexamer Ac-Pro(5)-t-BuPro NH(2) (4) produced a local 5-tert-butylprolyl amide population containing 61% cis isomer in water. In spite of the presence of a local prolyl cis amide geometry, the downstream prolyl amides in 4 remained in the trans isomer as determined by NOESY spectroscopy. Conformational analysis by (13)C-nmr and CD spectroscopy indicated that Ac-Pro(6)-NH(2) (3) adopted the all-trans amide polyproline type II helix in water. As the amount of 5-tert-butylproline increased from one to three residues in hexamers 4-6, a gradual destabilization of the polyproline type II helical geometry was observed by CD spectroscopy in water; however, no spectrum was obtained, indicative of a complete conversion to a polyproline type I helix. The implications of these results are discussed with respect to the previously proposed theoretical mechanisms for the helical interconversion of polyproline, which has been suggested to occur by either a cooperative C- to N terminal isomerization of the prolyl amide bonds or via a conformational intermediate composed of dispersed sequences of prolyl amide cis and trans isomers. PMID- 10679630 TI - Solution structure of nocistatin, a new peptide analgesic. AB - Nocistatin, a new heptadecapeptide encoded in the bPNP-3 gene, has a powerful biological activity connected with the mechanisms of pain transmission. It does not bind to the opioid receptors but to another brain receptor with high affinity. In order to substantiate these novel biological data with a structural basis, we have undertaken a conformational study in solution. Proton nmr data in helicogenic solvents are consistent with a well-defined helical structure that is consistent with the nmr parameters of the C-terminal octapeptide, a shorter fragment that retains allodynia-blocking activity. PMID- 10679631 TI - Electrostatic steering of substrate to acetylcholinesterase: analysis of field fluctuations. AB - Based on previous molecular dynamics simulation results for acetylcholinesterase dimer, we calculate and analyse the electrostatic field fluctuations around the enzyme. The results show that dynamic features of the electrostatic field favor attraction of the positively-charged substrate. An Internet link to an animation of the results is also provided. PMID- 10679633 TI - Members of the biospectroscopy editorial advisory board, 1995-1999 PMID- 10679632 TI - Hydroxyl group interactions in polysaccharides: a deuterium-induced differential isotope shift 13C-NMR investigation. AB - The secondary isotope shift in (13)C-nmr spectra in water was used to obtain information on the interactions of hydroxyl groups with their environment in polysaccharides. Specifically, the possibility of detecting the preference of intramolecular hydrogen bonding with respect to solvation was investigated. Different aliphatic alcohols were studied in both protic and aprotic solvents in order to obtain reference systems. The polysaccharides investigated were selected so as to include both different types of glycosidic linkages and different conformational properties of the polymeric chain. In addition to polysaccharides, beta-cyclodextrin and inulin were also investigated. The experiments demonstrated that isotope shift data can advantageously contribute to the understanding of the conformational properties of polysaccharides and in particular, in setting up of constraints in molecular modeling calculations. PMID- 10679634 TI - Raman spectroscopic study of polycrystalline mono- and polyunsaturated 1 eicosanoyl-d(39)-2-eicosenoyl-sn-glycero-3-phosphocholines: bilayer lipid clustering and acyl chain order and disorder characteristics. AB - Polycrystalline lipid samples of a series of mono- and polyunsaturated, double bond positional isomers of 1-eicosanoyl-d(39)-2-eicosenoyl-sn-glycero-3 phosphocholines [C(20-d(39)):C(20:1 Delta(j))PC, with j = 5, 8, 11, or 13; C(20 d(39)):C(20:2 Delta(11,14))PC; and C(20-d(39)):C(20:3 Delta(11, 14,17))PC] were investigated using vibrational Raman spectroscopy to assess the acyl chain packing order-disorder characteristics and putative bilayer cluster formation of the isotopically differentiated acyl chains. Perdeuteration of specifically the saturated sn-1 acyl chains for these bilayer systems enables each chain's intra- and intermolecular conformational and organizational properties to be evaluated separately. Various saturated chain methylene CD(2) and carbon-carbon (C&bond;C) stretching mode peak height intensity ratios and line width parameters for the polycrystalline samples demonstrate a high degree of sn-1 chain order that is unaffected by either the double bond placement or number of unsaturated bonds within the sn-2 chain. In contrast, the unsaturated sn-2 chain spectral signatures reflect increasing acyl chain conformational disorder as either the cis double bond is generally repositioned toward the chain terminus or the number of double bonds increases from one to three. The lipid bilayer chain packing differences observed between the sn-1 and sn-2 chains of this series of monounsaturated and polyunsaturated 20 carbon chain lipids suggest the existence of laterally distributed microdomains predicated on the formation of highly ordered, saturated sn-1 chain clusters. PMID- 10679635 TI - IR spectroscopic study of phospholipid emulsions containing cholesteryl oleate. AB - As models for the lipid organization of low density lipoproteins (LDL), protein free aqueous emulsions are prepared from dimyristoyl phosphatidyl choline (DMPC), dipalmitoyl phosphatidyl choline (DPPC), and cholesteryl oleate (CO). Aqueous dispersions containing these lipids are sonicated and yield stable particles with diameters varying between 20 and 40 nm as measured through electron microscopy. IR spectroscopy shows that emulsions consisting of DMPC, DPPC, and CO at 3/1/1 and 1/1/1 ratios undergo specific thermal transitions, depending on their composition, that can be assigned to the phospholipids forming the surface layer of the emulsion particles and to core-located CO. However, at the 1/3/1 DMPC/DPPC/CO ratio this lipid system exhibits an order-disorder transition of the mixed phospholipids with no significant transition associated with core-located CO. Observation of the methylene C&bond;H and C&bond;D stretching modes of nondeuterated and deuterated lipids enables the packing characteristics and conformational order of each lipid to be monitored separately. The transition temperature changes compared to the temperatures for the analogous transitions in neat CO and CO-free phospholipid vesicles suggest the existence of interactions between CO and the above phospholipids in the ternary emulsion particles; these interactions are stronger at the 1/3/1 DMPC/DPPC/CO ratio. The results show that interactions between core and surface phases are dependent on the emulsion lipid composition and that these findings may be extended to native lipoproteins. PMID- 10679636 TI - Keratin orientation in wool and feathers by polarized raman spectroscopy. AB - Good quality polarized Raman spectra of a single wool fiber and an intact feather barbule are presented. The intensity ratio of the alpha-helix component of the amide I band measured parallel and perpendicular to the wool fiber axis was 0.39 +/- 0.05. This is consistent with theoretical predictions based on orientational calculations using the normal Raman polarizability tensor for an alpha-helical amide I band where the protein strands are aligned roughly parallel with the fiber axis. However, the depolarized spectral intensity of the alpha-helix mode was greater than expected. For the feather barbule, despite high quality spectra, a unique orientation of the beta-sheet structure could not be determined using the Raman intensity ratios of the amide I band alone. Using previously developed methods, the protein chains were found to be oriented between 60 and 90 degrees from the long axis of the barbule compared to an angle of 51 degrees calculated from polarized IR spectra of the same barbule. The Raman tensor methods for the determination of protein orientation in these fibers was found to be constrained by the complexity of the materials and the limitations of the band fitting methods used to apportion the intensity among the various vibrational modes of their spectra. Other advantages and limitations of polarized Raman microscopic methods of structural determination are discussed. PMID- 10679637 TI - Characterization of equilibrium intermediates in denaturant-induced unfolding of ferrous and ferric cytochromes c using magnetic circular dichroism, circular dichroism, and optical absorption spectroscopies. AB - Protein unfolding during guanidine HCl denaturant titration of the reduced and oxidized forms of cytochrome c is monitored with magnetic circular dichroism (MCD), natural CD, and absorption of the heme bands and far-UV CD of the amide bands. Direct MCD spectral evidence is presented for bis-histidinyl heme ligation in the unfolded states of both the reduced and oxidized protein. For both redox states, the unfolding midpoints measured with MCD, which is an indicator of tertiary structure, are significantly lower than those measured with far-UV CD, an indicator of secondary structure. The disparate titration curves are interpreted in terms of a compound mechanism for denaturant-induced folding and unfolding involving a molten globulelike intermediate state (MG) with near-native secondary structure and nonnative tertiary structure and heme ligation. A comparison of the dependence of the free energy of formation of the MG intermediate on the redox state with the known contributions from heme ligation and solvation suggests that the heme is significantly more accessible to solvent in the MG intermediate than it is in the native state. PMID- 10679638 TI - Flow analysis for determination of paraoxon with use of immobilized acetylcholinesterase reactor and new type of chemiluminescent reaction. AB - A highly sensitive flow analysis method for determination of acetylcholinesterase (AChE) inhibitors like organophosphorous pesticides using a new chemiluminescent reaction was developed and optimized. This method is fast, sensitive, and cheap, because it requires only one enzyme and its substrate. The system incorporates a reactor with immobilized AChE on controlled pore glass (CPG) and a chemiluminometric detector. Variations in enzyme activity due to inhibition are measured from the changes of concentrations of thiocholine produced when the substrate (acetylthiocholine chloride) is pumped before and after the passage of the solution containing the pesticide through the immobilized AChE reactor. Thiocholine is determined by a new chemiluminescent reaction with luminol in the presence of potassium ferricyanide. The percentage inhibition of enzyme activity is correlated to the pesticide concentration. The inhibited enzyme is reactivated by 10 mM pyridine-2-aldoxime methiodide (2-PAM). The experimental conditions were first optimized for activity determination of the effect of pH, flow rates, and Tris concentrations. For the measurement of AChE inhibition, the appropriate concentration of the substrate is selected such that the rate of noninhibited reaction can be considered unchanged and could be used as a reference. For optimization of experimental conditions for inhibition, several parameters of the system are studied and discussed: flow rate, enzyme-pesticide contact time, luminol concentration, ferricyanide concentration, 2-PAM concentration, and configuration of the FIA manifold. Paraoxon, an organophosphorous pesticide was tested. For an inhibition time of 10 min the calibration graph is linear from 0.1 to 1 ppm paraoxon with a relative standard deviation (n = 5) of 4.6% at 0.5 ppm. For an inhibition time of 30 min the calibration graph is linear from 25 to 250 ppb paraoxon. PMID- 10679639 TI - IR study of self-assembly of capsular exopolymers from Pseudomonas sp. NCIMB 2021 on hydrophilic and hydrophobic surfaces. AB - Capsular exopolymers (EPS) of the bacterium Pseudomonas sp. NCIMB 2021 are allowed to self-assemble on hydrophilic and hydrophobic gold surfaces. Tapping mode atomic force microscopy confirms the differences in the surface topography between EPS adsorbed on both surfaces. Fourier-transform IR spectroscopy indicates that the EPS surface coverage is much greater on the hydrophobic surface. Furthermore, an increased contribution is observed from hydrophobic (i.e., methyl and tyrosyl residues) and electrostatic (i.e., carboxylate residues) groups at the hydrophobic surface, but there is relatively less neutral polymer compared to the hydrophilic surface. The behavior of this EPS is in agreement with the behavior of cells of Pseudomonas sp. NCIMB 2021 at hydrophilic and hydrophobic surfaces. PMID- 10679640 TI - Adrenocortical carcinoma: clinical and laboratory observations. AB - BACKGROUND: The clinical features and natural history of adrenocortical carcinoma are highly dependent on the type of center reporting their experience. Observations from oncology services suggest a high incidence of nonfunctioning tumors, whereas reports from endocrine clinics emphasize excessive corticoid and androgen production in the majority of tumors. The incidence rate and natural history of childhood adrenal carcinoma generally has been under emphasized. METHODS: Over the past 17 years, the authors have evaluated and treated 47 patients with adrenocortical carcinoma referred to the University of Sao Paulo, 22 of whom were children. RESULTS: There is a bimodal age incidence of adrenal carcinoma, with the disease peaking in the first and fourth decades of life. Childhood adrenal carcinoma is characterized by a high rate of incidence of virilization, marked overproduction of androgens, and a less aggressive clinical course, and appears to be more amenable to surgical and other therapeutic modalities. By contrast, adrenocortical carcinoma occurring in adults presents more commonly as a mixed Cushing and virilizing syndrome, with overproduction of corticoids and androgens and a far more aggressive clinical course, leading to rapid death within months or years. Nonfunctioning adrenocortical carcinoma is less common; it generally occurs in older adults and exhibits a rapid downhill course. Modern day imaging methods have improved the diagnosis and staging of adrenal carcinoma greatly. In the authors' experience, the histologic criteria of Weiss appeared to predict tumor prognosis most accurately, whereas immunologic markers, cytoskeletal markers, DNA ploidy, cell phase markers, and oncogenic probes have yielded inconsistent results to date. Surgical removal of a localized tumor remains the best hope for long term survival. Medical therapy with mitotane and its successors in patients with Stage III or IV (MacFarlane system as modified by Sullivan et al.) disease appear to have added little to longevity or quality of life. CONCLUSIONS: When diagnosed in children, adrenal carcinoma is associated with virilism and a less aggressive natural history; however, when it occurs in adults, the disease presents more commonly as a mixed Cushing virilizing syndrome and has a virulent course. The Weiss histologic criteria appear to correlate best with disease prognosis, but other histochemical, cell cycle, and genetic markers have not, to date, aided in disease management. PMID- 10679641 TI - Vitamins, carotenoids, dietary fiber, and the risk of gastric carcinoma: results from a prospective study after 6.3 years of follow-up. AB - BACKGROUND: Numerous components of fruit and vegetables are considered to decrease the risk of gastric carcinoma. In the current prospective study, the authors examined the association between the intake of vitamins, carotenoids, and dietary fiber and vitamin supplement use and the incidence rate of gastric carcinoma. METHODS: The Netherlands Cohort Study began in 1986 with 120,852 men and women ages 55-69 years. Data regarding diet and other covariates were collected by means of a self-administered questionnaire. After 6.3 years of follow-up, data regarding 282 incident cases of gastric carcinoma and 3123 subcohort members were available for case-cohort analyses. RESULTS: In analyses adjusted for age, gender, smoking history, education, stomach disorders, and family history of gastric carcinoma, an inverse association with vitamin C intake (relative risk [RR] for highest vs. lowest intake category, 0.7; 95% confidence interval [95% CI], 0.5-1.0) was observed, with a borderline significant trend across three intake categories (P = 0. 06). After the exclusion of cases diagnosed in the first and second follow-up years, the RR was 0.9 (95% CI, 0.6 1.2; P trend = 0.44). Intake of retinol and beta-carotene were associated positively with gastric carcinoma risk with highest versus lowest quintile RRs of 1. 6 (95% CI, 1.0-2.5; P trend = 0.02) and 1.6 (95% CI, 1.0-2.6; P trend = 0.13), respectively, after the exclusion of first-year and second-year cases. Intake of folate, vitamin E, alpha-carotene, lutein plus zeaxanthin, beta-cryptoxanthin, lycopene, and dietary fiber was not associated with gastric carcinoma. Patients who used vitamin A-containing supplements had a lower risk of gastric carcinoma than nonusers (RR = 0.4; 95% CI, 0.2-0.9). CONCLUSIONS: No clear inverse associations were found between the intake of vitamins, carotenoids, and dietary fiber and the risk of gastric carcinoma after adjustment for confounding variables and the exclusion of first-year and second-year cases. PMID- 10679642 TI - Molecular identification of metastatic cancer to the skin using laser capture microdissection: a case report. AB - BACKGROUND: In the current study the authors report a 57-year-old woman with a scalp tumor and cervical lymphadenopathy who had a previously resected duodenal carcinoid. Histologic and immunophenotypic characteristics of the duodenal carcinoid differed from those of the scalp and cervical lymph node tumors, prompting the use of molecular methodologies to make the diagnosis. METHODS: Paraffin embedded tissues from the duodenal carcinoid, scalp, and lymph node tumors were dissected using microscopic visualization and laser capture microdissection. DNA was extracted and polymerase chain reaction (PCR) was performed to evaluate loss of heterozygosity and microsatellite alterations using primers flanking 22 polymorphic microsatellite markers from 9 chromosomal regions, including genes associated with MEN-1 (11q), CDKN2 (9p), p53 (17p), and bronchial carcinoid (3p). Microdissected lymphocytes from the three tissues were used as source of constitutional DNA (controls). RESULTS: Fourteen of the 22 markers were informative (heterozygous in control lymphocytes). A marker on 3p12 showed loss of the same parental allele in the three tumors. A different marker on 3p14.2 showed an identical shifted band in the three tumors indicative of a common microsatellite alteration. CONCLUSIONS: The shared molecular abnormalities among the three tumors indicated a common clonal origin, leading to a diagnosis of primary duodenal carcinoid with clear cell metastases to the scalp and cervical lymph nodes. These findings led to radiation therapy and immunotherapy rather than chemotherapy. This case illustrates the novel application of laser capture microdissection combined with PCR-based analyses of genomic markers for the identification of the origin of metastatic disease. PMID- 10679643 TI - Adenomatous polyposis coli I1307K mutation in Jewish patients with different ethnicity: prevalence and phenotype. AB - BACKGROUND: A new mutation, I1307K, recently was reported in the adenomatous polyposis coli (APC) gene. This mutation was found to be predominant in Ashkenazi Jews, creating a hypermutable area and predisposing the development of carcinoma. The objective of the current study was to estimate the prevalence of this mutation in several of the ethnic groups that comprise the Israeli population and to elucidate the clinical features of the mutation carriers with colorectal carcinoma (CRC). METHODS: A total of 111 consecutive CRC patients were evaluated and their medical history and clinical data recorded. The general population (298 Ashkenazim and 189 Yemenites) also was tested for the presence of this mutation. Mutation screening was performed using both the polymerase chain reaction-based amplification refractory mutation system and a commercial APC kit. RESULTS: Of the total of 111 CRC patients, 15 (13.5%) carried the I1307K mutation and 26 of 487 subjects from the general population (5.3%) carried the I1307K mutation (P = 0.004). Among the 71 Ashkenazi CRC patients there were 12 carriers (16.9%) whereas 17 of the 298 Ashkenazi Jewish general population (5.7%) carried the mutation (P = 0.004). Of the 4 CRC patients of Yemenite origin, 3 carried the mutation and 9 carriers were found among 189 subjects in the general Yemenite population (4.7%) (P = 0.0007). None of the 34 Sepharadic or 2 Arab CRC patients carried the APC I1307K allele. Late age at diagnosis (64.6 years +/- 10.0, which is similar to that of the noncarriers), mostly right-sided tumors, and moderate to good differentiation constituted the phenotype of the mutation carriers. CONCLUSIONS: The authors believe the findings of the current study broaden the known spectrum of ethnic groups in which the APC I1307K mutation is prevalent. The phenotype of the mutation carrier CRC patients does not conform to the expected familial pattern of germline mutations. The phenotype and the differential incidence rate of CRC among APC I1307K carriers of various ethnic groups suggest low penetrance. PMID- 10679644 TI - Abnormal expression of hepatoma specific gamma-glutamyl transferase and alteration of gamma-glutamyl transferase gene methylation status in patients with hepatocellular carcinoma. AB - BACKGROUND: Hepatoma specific gamma-glutamyl transferase (HS-GGT) bands were expressed in the development of hepatocellular carcinoma (HCC) and were associated with a high incidence of HCC diagnosis. The objectives of this study were to determine the levels of HS-GGT quantitatively in the sera of patients with different liver diseases. The methylational status of GGT gene CCGG sites was analyzed in hepatoma tissues. METHODS: The HS-GGT concentrations were quantitatively analyzed in the sera of 156 HCC patients and others with liver diseases or extrahepatic tumors. In 20 hepatoma tissues, the GGT enzyme proteins were purified, the activities of GGTs of different molecular form were examined, total RNAs were extracted and amplified by using a nested polymerase chain reaction (PCR) assay, and the methylational status of CCGG site (M3) in the 5' noncoding region of GGT genes was investigated with the restriction enzyme Hpa II. RESULTS: Total GGT activities in patients with liver diseases and extrahepatic tumors were abnormally increased. The levels of serum HS-GGT were significantly elevated (P < 0.001) in the HCC group; the incidence of HS-GGT over 5.5 IU/L was 86% in HCC patients and less than 3% in patients with other diseases. From liver cancer to distal noncancerous tissues, an increasing tendency (P < 0.05) of total RNA concentrations was found; the frequencies of amplified fragment and hypomethylated M3 site of GGT genes were 100% and 75% in HCC, 85% and 55% in paracancerous tissues, and 75% and 50% in noncancerous tissues, respectively. An inverse correlation was found between methylational degrees of GGT genes and expression levels of GGT. CONCLUSIONS: The abnormal alteration of serum HS-GGT level is a sensitive tumor marker for HCC diagnosis or differentiation, and the overexpression of GGT in HCC may be related to the hypomethylational status of CCGG sites of GGT genes. PMID- 10679645 TI - Treatment of carcinoid syndrome: a prospective crossover evaluation of lanreotide versus octreotide in terms of efficacy, patient acceptability, and tolerance. AB - BACKGROUND: The somatostatin analogues lanreotide and octreotide have previously been shown to be effective in controlling flushing and diarrhea in patients with carcinoid syndrome. As lanreotide requires injection only every 10 days, compared with twice-daily injections of octreotide, a direct comparison between these two treatments in terms of patient acceptability, patient preference, and efficacy in controlling symptoms was performed in patients with carcinoid syndrome. METHODS: Thirty-three patients with carcinoid syndrome were included in an open, multicenter, crossover study. Half of the patients received octreotide 200 microg subcutaneously twice or thrice daily for 1 month followed by lanreotide 30 mg intramuscularly every 10 days for 1 month, while the other half commenced with lanreotide followed by octreotide in a similar fashion. Quality-of-life assessments were performed at each visit and patient preference for one of the two treatments evaluated. The number and intensity of flushing episodes and bowel movements, urinary 5-hydroxyindoleacetic acid (5HIAA) levels, and plasma serotonin levels were recorded. RESULTS: No significant differences were found between lanreotide and octreotide in terms of quality of life. The majority of patients (68%) preferred lanreotide (P = 0.03), largely due to its simplified mode of administration. Disappearance or improvement in flushes occurred in 53.8% of patients (14 of 26) while on lanreotide and in 68% (17 of 25) on octreotide. A disappearance or improvement of diarrhea in 45.4% (10 of 22) on lanreotide, compared with 50% (11 of 22) on octreotide, was also observed. Lanreotide and octreotide were equally effective in reducing urinary 5HIAA levels and plasma serotonin levels. Both treatments were well tolerated, with mild symptoms of abdominal pain and nausea observed in 29% and 14% receiving octreotide and lanreotide, respectively. CONCLUSIONS: Lanreotide and octreotide are equally efficacious in terms of symptom control and reduction in tumor cell markers for patients with carcinoid syndrome. Due to its simplified mode of administration, most patients prefer treatment with lanreotide. PMID- 10679646 TI - Current follow-up strategies after potentially curative resection of extremity sarcomas: results of a survey of the members of the society of surgical oncology. AB - BACKGROUND: The follow-up of patients after potentially curative resection of extremity sarcomas has significant clinical and fiscal implications. However, the ideal postoperative surveillance regimen for these uncommon neoplasms remains ill defined. This study was designed to determine the current follow-up practices of a large, diverse group of physicians who care for sarcoma patients. METHODS: The 1592 members of the Society of Surgical Oncology (SSO) were surveyed regarding their follow-up practices with a detailed questionnaire mailed in 1997. Information regarding frequency of follow-up testing was requested for extremity sarcoma patients treated for cure based on 4 vignettes: low grade lesion 5 cm and high grade lesion 5 cm. Respondents were asked to indicate the number of office visits, laboratory tests and imaging studies performed annually during the first 5 years and the 10th year after surgery. RESULTS: Forty-five percent (716 of 1592) completed the survey. Of the 343 respondents who performed sarcoma surgery, 318 (93%) also provided long term postoperative follow-up for their patients. Ninety-four percent of respondents (295 of 318) were trained in general surgery and 5% (15 of 318) completed orthopedic residencies. Ninety-one percent (291 of 318) were also fellowship trained (80% in surgical oncology). Sixty-three percent (201 of 318) were in academic practice. Routine office visits and chest X-ray (CXR) were the most frequently performed items for each of the years. The frequency of office visits and CXR increased with tumor size and grade and decreased with postoperative year. Complete blood count and liver function tests were the most commonly ordered blood tests, but many respondents did not order any blood tests routinely. Imaging studies of the extremities were performed on the majority of patients with large (> 5 cm) low grade lesions and on both large and small high grade lesions during the first postoperative year. CONCLUSIONS: Postoperative sarcoma surveillance strategies utilized by members of the SSO rely most heavily on office visits and CXR. Tumor grade, tumor size, and postoperative year affect surveillance intensity. PMID- 10679647 TI - Menstrual and reproductive factors and risk of soft tissue sarcomas. AB - BACKGROUND: Soft tissue sarcomas (STS) are a heterogeneous group of neoplasms whose etiology remains largely undefined. A role for female hormones in the development of STS has been suggested. To investigate this possibility, the authors analyzed data from a hospital-based case-control study conducted in Northern Italy between 1983 and 1998. METHODS: Cases were 104 women aged < 79 years with incident STS who were admitted to the cancer institutes and major teaching and general hospitals. Controls were 505 women admitted to the same network of hospitals for acute, nonneoplastic, nongynecologic, and nonimmune related conditions. RESULTS: The multivariate odds ratio (OR) for women aged >/= 15 years compared with those aged < 12 years at menarche was 1.94 (95% confidence intervals [95% CI], 0.80-4.74). No association with STS risk was observed for menstrual cycle pattern, age at menopause, parity, and abortions. Late age at first pregnancy and birth were found to be related to an increased risk of STS, with an OR of 3.16 (95% CI, 0. 96-10.44) and 2.79 (95%% CI, 0.79-9.90) for women aged >/= 30 years at first pregnancy and birth compared with those aged < 20 years. The trend in risk was significant for age at first pregnancy. No relation with the risk of STS emerged for age at last birth and time since first or last birth. CONCLUSIONS: The risk of STS was found to be weakly related to late age at first pregnancy or birth, but not to other menstrual and reproductive factors. PMID- 10679648 TI - High dose chemotherapy and autologous blood stem cell support in women with breast carcinoma and isolated supraclavicular lymph node metastases. AB - BACKGROUND: The prognosis of patients with isolated supraclavicular lymph node (SCN) metastases is similar to patients with metastatic breast carcinoma involving other sites. Because these patients have a lower disease burden compared with women with distant metastases, their outcome after high dose chemotherapy (HDCT) may be superior. METHODS: The authors evaluated event free survival (EFS) and overall survival in a series of 20 consecutive women with SCN metastases as the only site of metastatic disease who were treated with HDCT and peripheral blood stem cell transplantation at The Toronto Hospital. All patients had responded to 4-6 cycles of induction CT using either an anthracycline containing regimen or a single agent taxane, and received intensive therapy comprised of mitoxantrone, 64 mg/m(2); cyclophosphamide, 6000 mg/m(2); and carboplatin, 800-2000 mg/m(2), each divided over 4 days followed by the infusion of autologous peripheral blood stem cells. Involved field radiation therapy (RT) was administered when possible after transplantation to the supraclavicular fossa and tamoxifen was given to previously untreated patients if they were hormone receptor positive or if their hormone receptor status was unknown. RESULTS: At a median follow-up of 28 months, 13 of the 20 women were alive, 11 of whom (55%) remained in continuous complete remission. There were no treatment-related deaths. The median overall survival was 37 months and the median progression free survival was 32 months from the date of transplantation. Consolidative RT was delivered to 11 women and on univariate analysis was found to be significantly associated with better EFS (P = 0.02). CONCLUSIONS: The long term outcome of women with breast carcinoma and isolated SCN metastases whose disease is sensitive to CT appears to be favorable; whether this result is superior to that achieved with standard therapy alone remains to be confirmed in prospective, randomized trials. PMID- 10679649 TI - The predictive value of body protein for chemotherapy-induced toxicity. AB - BACKGROUND: The use of body surface area in determining chemotherapy dosing, particularly in the obese, remains controversial. Total body nitrogen (TBN) measurement in patients with serious illness has been suggested to be an accurate predictor of clinical course. The ability of TBN to predict chemotherapy-induced neutropenia was examined in the current study. METHODS: TBN measurements were performed in 31 female outpatients with breast carcinoma who were undergoing standard cyclophosphamide, methotrexate, and 5-fluorouracil (CMF)-based chemotherapy (median age, 48 years; range, 26- 77 years). TBN was measured using the in vivo neutron capture analysis technique on Day 1 of Cycles 2-6. The chemotherapy toxicity index used was the absolute neutrophil count nadir (ANCN). Neutropenia was defined as an ANCN < 1.0 x 10(9)/L. The nitrogen index (NI) (TBN expressed as a percentage of age-, gender-. and height-matched healthy patients) then was compared with the corresponding ANCN values. RESULTS: Using receiver operating characteristics analysis, a "cut-off" value of NI = 0.89 was found. In this group of patients, when the NI was < 0.89, 11 of 13 courses in 7 patients (85%) led to an ANCN of < 1.0 x 10(9)/L, and when the NI was > 0.89, 29 of 109 courses (27%) led to an ANCN of < 1.0 x 10(9)/L (P < 0.0001). CONCLUSIONS: In this small group of breast carcinoma patients, the NI was found to be the most powerful predictor of neutropenia after CMF-based chemotherapy. The authors conclude that NI may be a useful clinical tool in identifying patients at a higher risk of chemotherapy-induced toxicity when widely distributed drug combinations such as CMF are used, and warrants further study with other commonly used drugs or drug regimens. PMID- 10679650 TI - The prognostic value of p53 and c-erb B-2 immunostaining is overrated for patients with lymph node negative breast carcinoma: a multivariate analysis of prognostic factors in 613 patients with a follow-up of 14-30 years. AB - BACKGROUND: Approximately 30% of breast carcinoma patients with negative lymph nodes die of their disease. Biologic markers such p53 protein and c-erb B-2 have been related to tumor progression, but their prognostic value remains controversial. METHODS: Two large series of a total of 613 lymph node negative breast carcinoma patients from a single institution were analyzed with respect to tumor size, histologic grade, and immunohistochemical staining for p53, c-erb B 2, estrogen receptor (ER), and progesterone receptor (PgR). Interobserver variation in histologic grading was evaluated by Kappa statistics. The two series had different treatment modalities: 228 patients (SACGS group) were treated surgically with mastectomy and given 1 perioperative chemotherapy course, and 385 patients (HOST group) were treated with mastectomy and ovarian radiation and further randomized to receive postoperative treatment with radiotherapy or no adjuvant treatment. The follow-up ranged from 14-30 years. RESULTS: Immunoreactivities for p53, c-erb B-2, ER, and PgR did not differ significantly in the two series. p53 immunostaining was present in 187 of 613 tumors (29%), and c-erb B-2 immunoreactivity was present in 58 of the tumors (10%). Three hundred forty-eight tumors (57%) were positive for ER. Kappa statistics value of interobserver variation in the histologic grading of ductal carcinomas was 0.69, which is considered to be a substantial degree of agreement. No significant differences in survival were found when comparing p53, c-erb B-2, ER, and PgR positive and negative cases. However, both recurrence free survival rates and overall survival rates after 10 years were significantly better in the T1N0M0 group compared with the T2N0M0 group (81% vs. 67% [P < 0.0001] and 85% vs. 70% [P < 0.0001]). Ten-year recurrence free survival rates for patients with histologic Grade 1 versus Grades 2-3 (according to Elston and Ellis' modification of the Bloom and Richardson method) tumors were 90% and 70%, respectively (P < 0. 0001), and overall survival rates for the same groups were 94% and 81%, respectively (P=0.0002). After 30 years of follow-up, the overall survival rate for patients with tumors of histologic Grade 1 versus Grades 2-3 were 87% and 68%, respectively, and were 78% and 66%, respectively, for patients with tumors 20-50 mm. Approximately 35% of the patients with tumors of histologic Grades 2-3 and measuring > 20 mm were dead after 10 years of follow-up, contrary to 6% of the patients with tumors of histologic Grade 1 measuring /=T3) are at risk of recurrence, metastasis, and dying of the disease, whereas patients in this study with T1 or T2 disease had favorable outcomes after complete tumor resection. PMID- 10679655 TI - Atypical nephrogenic metaplasia of the urinary tract: a precursor lesion? AB - BACKGROUND: Nephrogenic metaplasia with cytologic atypia (atypical nephrogenic metaplasia) is occasionally encountered and its biologic potential is uncertain. METHODS: The authors describe 18 cases of atypical nephrogenic metaplasia characterized by the presence of prominent cytologic atypia, including nuclear enlargement, nuclear hyperchromasia, and enlarged nucleoli. DNA ploidy analysis by digital image analysis and immunostaining for high-molecular-weight cytokeratin (34betaE12), cytokeratin 7, cytokeratin 20, carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), p53, and MIB-1 were performed in 9 cases. RESULTS: The mean patient age was 62 years (median, 65 years; range, 39-84 years). The male-to-female ratio was 2.6:1. Two patients had a history of noninvasive papillary urothelial carcinoma. The typical clinical presentation was hematuria (8 patients) and voiding symptoms (5 patients). Cystoscopic findings were suspicious for neoplasm in 7 of 13 cases. The neoplastic cells were positive for high-molecular-weight cytokeratin, cytokeratin 7, and EMA, and were usually negative for cytokeratin 20 and CEA. p53 nuclear accumulation and increased MIB-1 labeling index were seen in 4 cases. DNA ploidy analysis showed aneuploid pattern in 2 of 9 cases. The mean patient follow-up was 3.5 years (range, 0.5-10.6 years); 2 patients had recurrent nephrogenic metaplasia, and the remainder were alive without recurrence or urothelial carcinoma. CONCLUSIONS: Atypical nephrogenic metaplasia is benign; it occasionally displays substantial cytologic abnormalities of no apparent clinical significance. Awareness of the spectrum of cytologic changes within this entity is critical to prevent overdiagnosis of cancer and avoid unnecessary treatment. There is no direct evidence that links atypical nephrogenic metaplasia to cancer. PMID- 10679656 TI - Evaluation of cell proliferation, epidermal growth factor receptor, and bcl-2 immunoexpression as prognostic factors for patients with World Health Organization grade 2 oligodendroglioma. AB - BACKGROUND: Prognostic factors in oligodendrogliomas are an area of controversy in neuropathology. Although grading and the study of some morphologic variables may be of value as prognostic parameters, the variability of postoperative disease free survival in patients with World Health Organization Grade 2 oligodendroglioma indicates that the biologic behavior of this entity remains unknown. The objective of the current study was to evaluate immunoexpression of the proliferation index (PI), epidermal growth factor receptor (EGFR), and bcl-2 as prognostic factors in patients with Grade 2 oligodendroglioma. METHODS: In a series of 19 cases of pure Grade 2 oligodendroglioma, we assessed the mitotic count, labeling index for MIB-1 and PCNA, and immunoreactivity for EGFR and bcl-2 with semiquantitative parameters and compared these with postoperative disease free survival. Statistical analyses using the Cox-Mantel nonparametric test and Spearman correlation coefficient were used to evaluate the data. RESULTS: Disease free survival was significantly shorter when the MIB-1 PI was > 5% (P = 0.0096) and the PCNA PI was > 9% (P = 0.00011) and when mitoses were observed (P = 0.00004). The paired variables also were found to correlate: MIB-1 versus PCNA (P = 0.04), MIB-1 versus mitotic count (P = 0.0184), and PCNA versus mitotic count (P = 0.0079). In all cases, there was immunoreactivity for EGFR; conversely, all cases were negative for bcl-2 in the cells with an oligodendrocyte phenotype. CONCLUSIONS: The PI was found to correlate with the postoperative disease free survival in patients with Grade 2 oligodendroglioma; therefore, the authors suggest a possible subdivision of Grade 2 oligodendrogliomas into two groups based on the mitotic count and PI. PMID- 10679657 TI - Stereotactic radiosurgery for recurrent ependymoma. AB - BACKGROUND: Patients with recurrent intracranial ependymomas were evaluated to assess local control, overall survival, and complications from stereotactic radiosurgery (SRS). METHODS: Twelve patients (with a total of 17 tumors) with recurrent ependymoma underwent SRS. Local failure was defined as tumor progression within the prescription isodose volume, and marginal failure was defined as tumor progression adjacent to the SRS prescription isodose volume. Tumor progression away from the prescription volume was considered distant failure. Eleven of the 12 patients had undergone previous resection and external beam radiation therapy (46-56 Grays [Gy]; median, 54 Gy) before radiosurgery, and 1 patient had failure after complete resection alone. Age at SRS ranged from 5-56 years (median, 29 years). Three patients were female. The marginal tumor dose was 12-24 Gy (median, 18 Gy). One to 14 isocenters (median, 4 isocenters) were utilized to irradiate volumes of 0.3-15.5 cm(3) (median, 3.2 cm(3)). RESULTS: The duration of follow-up ranged from 2.5-60 months (median, 22.5 months). The median overall survival after SRS was 3.4 years (range, 1.4-5 years). In-field local control was achieved in 14 of the 17 tumor sites and estimated 3-year local control was 68%. There were two in-field failures and one marginal failure. Distant failure occurred in two patients. Two patients developed treatment related complications after SRS. CONCLUSIONS: SRS provides good local tumor control for patients with recurrent intracranial ependymoma and may have a favorable impact on survival. SRS should be evaluated more extensively in the initial treatment of patients with ependymoma to minimize local failure after surgical management. PMID- 10679658 TI - Mature results of a phase III randomized trial comparing concurrent chemoradiotherapy with radiation therapy alone in patients with stage III and IV squamous cell carcinoma of the head and neck. AB - BACKGROUND: The current study presents mature results from a Phase III randomized trial comparing radiation therapy and concurrent chemoradiotherapy in patients with resectable American Joint Committee on Cancer Stage III and IV disease. METHODS: One hundred patients were randomized to receive either radiation therapy alone (Arm A) (at a dose of between 66-72 grays [Gy] at 1.8-2 Gy per day) and the identical radiation therapy with concurrent chemotherapy (Arm B) (5-fluorouracil, 1000 mg/m(2)/day, and cisplatin, 20 mg/m(2)/day, both given as continuous intravenous infusions over 4 days beginning on Days 1 and 22 of the radiation therapy). Primary site resection was planned for patients with residual or recurrent local disease. Cervical lymph node dissection was performed for regional persistent disease or recurrence, or if N2-3 disease was present at the time of presentation. RESULTS: After completing all therapy including surgery, 82% of the patients in Arm A and 98% of the patients in Arm B had been rendered disease free (P = 0.02). At a median follow-up of 5 years (range, 3-8 years), the 5-year Kaplan-Meier projections for overall survival for Arm A versus Arm B were 48% versus 50% (P = 0.55). Kaplan-Meier projections for the recurrence free interval were 51% versus 62% (P = 0.04), projections for a distant metastasis free interval were 75% versus 84% (P = 0. 09), projections for overall survival with primary site preservation were 34% versus 42% (P = 0.004), and projections for local control without surgical resection were 45% versus 77% (P < 0.001). Salvage surgery proved to be successful in 63% and 73%, respectively, of the Arm A and Arm B patients with primary site failure. Unrelated death while free of disease occurred in 22% and 32%, respectively, of Arm A and Arm B patients (P = 0.26). CONCLUSIONS: The addition of concurrent chemotherapy to definitive radiation in patients with resectable Stage III and IV squamous cell carcinoma of the head and neck improves the likelihood of disease clearance, a recurrence free interval, and primary site preservation. However, overall survival does not appear to be improved, reflecting both effective surgical salvage after local recurrence and competing causes of death. PMID- 10679659 TI - The origins of multiple squamous cell carcinomas in the aerodigestive tract. AB - BACKGROUND: Chemoprevention and cessation of smoking and alcohol may prevent development of multiple tumors (MTs) in the aerodigestive tract if new MTs arise independently, but they are of no benefit if MTs are due to migration of an already transformed clone of tumor cells. This issue was addressed in this study by investigation of the clonality among MTs. METHODS: Mutation analysis of the entire coding region of p53 and loss of heterozygosity (LOH) pattern analysis of microsatellite markers on chromosome arms 3p, 9p, and 17p are promising for the investigation of clonality. In the first part of this study, the authors established the variability and stability of these clonal markers by comparing primary head and neck squamous cell carcinomas (HNSCCs) with their metastases. In the second part of this study, the authors evaluated nine patients with multiple HNSCCs using these markers. In the final part, the authors illustrate the use of these clonal markers in 11 patients for whom there was diagnostic uncertainty as to whether their second squamous cell carcinoma was either a new primary tumor, a metastasis, or a recurrence. RESULTS: Both p53 gene mutations and LOH patterns were stable during tumor progression. Furthermore, the variability of p53 gene mutations was high. More than 90% of the tumors contained a p53 mutation. A particular mutation never occurred more than twice in a total of 69 primary HNSCCs. Five of 69 cases presented a common mutation. In contrast, LOH patterns showed less variability; they were identical in 5 of 16 cases. The metachronous HNSCCs from nine patients all showed different p53 mutations, and in the three cases that were subjected to LOH analysis different patterns were observed. All 11 patients for whom there was diagnostic uncertainty about the origin of their second squamous cell carcinoma could be categorized as having multiple primary tumors, disseminated disease, or recurrent disease. CONCLUSIONS: Metachronous HNSCCs at different locations are not clonally related and thus have not developed from the migration of tumor cells. PMID- 10679660 TI - Bone marrow staging of patients with non-Hodgkin lymphoma by flow cytometry: correlation with morphology. AB - BACKGROUND: Immunophenotypic analysis is an established tool in the diagnosis and classification of many hematolymphoid disorders; however, the role of flow cytometry (FC) in detecting bone marrow involvement during the staging of non Hodgkin lymphoma (NHL) has yet to be defined. METHODS: The authors retrospectively analyzed 157 staging and 70 restaging bone marrow biopsies on which morphologic and FC analyses were performed; these biopsies were taken from 195 consecutive patients. Bone marrow biopsies were blindly and independently reviewed and determined to be positive, negative, or suspicious for morphologic involvement by NHL, with disagreements settled by a third reviewer. A selected panel of monoclonal antibodies was used to determine whether bone marrow involvement was immunophenotypically positive (>5%), minimal (<5%), negative, or nondiagnostic. RESULTS: FC and morphology agreed in 78% of cases (178 of 227: 129 both negative, 49 both positive) and were discrepant in 22% (49 of 227). Seven percent (16 of 227) were morphologically positive but showed no evidence of disease on FC, whereas 12% (27 of 227) were positive by FC but had no morphologic involvement. Of the 162 morphologically negative or suspicious bone marrows, 27 were shown to be involved by FC, resulting in a false-negative detection rate of 17%. Most of these (22 of 27, 81%) had minimal detectable disease. Seven percent of Stage I and 26% of Stage II NHL cases with negative staging bone marrow morphologically were found to be involved by FC. CONCLUSIONS: Neither morphologic examination of bone marrow biopsy specimens nor FC alone is adequate to detect all cases of NHL with bone marrow involvement. FC is most sensitive for detecting minimal bone marrow lymphoma, whereas morphology will detect most cases in which involvement is >5%. Cases of early stage NHL with morphologically negative bone marrow could potentially be restaged as Stage IV on the basis of FC results. The clinical importance of minimal bone marrow involvement by NHL needs further evaluation. PMID- 10679661 TI - Assessment of the need for palliative care as perceived by individual cancer patients and their families: a review of instruments for improving patient participation in palliative care. AB - BACKGROUND: Palliative care should fit the needs of individual cancer patients and their families. Instruments can help to improve needs assessment. This review evaluates questionnaires for the systematic assessment of needs experienced by individual cancer patients for help, care, or support, as well as the needs of their family members. METHODS: The MEDLINE and PsycLIT data bases were searched systematically. Questionnaires were evaluated by reviewing their contents and estimating their validity, reliability, and feasibility for use in caregiving practice. RESULTS: Analysis of the 471 articles identified from the searches yielded 9 questionnaires for the assessment of patients' needs and 6 questionnaires for the assessment of family members' needs. Most of these instruments were carefully constructed; their validity and reliability were satisfactory and well documented. However, in most questionnaires the needs for care were confounded by satisfaction with care, and the problems experienced by patients. Only one questionnaire for patients specifically addressed the need for help; none for family members was so specific. Data on the feasibility of questionnaires for use in regular care were scarce. Issues frequently omitted were spiritual issues, the personal needs of family members, and the continuity of care. CONCLUSIONS: None of the instruments the authors found was complete for all dimensions of palliative care. Most instruments were constructed for research purposes and had not been tested for use in practical care. Further development of practical instruments based on a theoretical concept of needs assessment seems necessary. The feasibility of instruments for use in practical caregiving and their effects on the quality of care needs further research. PMID- 10679662 TI - The National Cancer Data Base Report on treatment patterns for hepatocellular carcinomas: improved survival of surgically resected patients, 1985-1996. AB - BACKGROUND: The Commission on Cancer data from the National Cancer Data Base (NCDB) has previously reported data evaluating time trends in various cancers, including such elements as stage of disease at diagnosis, treatment, and survival for multiple tumor sites. In this report, data collected from 1985, 1986, 1990, 1991, 1995, and 1996 for primary hepatocellular carcinoma (HCC) tumors are presented. METHODS: The data presented in this review were collected from hospital cancer registries from across the U.S. Eight calls for data have yielded a total 6.9 million cases for the years 1985-1996, including 1158 HCC cases in 1985-1986, 3319 cases in 1990-1991, and 5683 cases in 1994-1995 from hospital cancer registries across the U. S. These data represent approximately 4.3%, 11.2%, and 14.8% of the estimated cases of carcinomas of the liver and biliary tract diagnosed in the U.S. in each of the three respective time periods. RESULTS: The outcome for patients diagnosed with HCC remains poor, with only 10% of patients with American Joint Committee on Cancer Stage I disease surviving 5 years. Approximately 50% of patients received no therapy for their HCC, even those with early stage disease. Over these three time periods, the use of chemotherapy appears to have decreased. Among patients diagnosed with Stage II and III disease a difference in survival was noted between those treated with surgery only and those treated with chemotherapy only. Women appear to have a limited survival advantage over men. CONCLUSIONS: In spite of an overall poor prognosis, subsets of patients with HCC appear to benefit from surgical resection/ablation of their tumor. The decreasing use of chemotherapy and the early reports of newer ablative techniques (e.g., cryotherapy) suggest that other treatment modalities are emerging. These NCDB data provide a baseline for HCC treatment from which prospective studies are being developed to assess the newer treatments as well as the underlying causes. PMID- 10679663 TI - The National Cancer Data Base Report on poor survival of U.S. gastric carcinoma patients treated with gastrectomy: Fifth Edition American Joint Committee on Cancer staging, proximal disease, and the "different disease" hypothesis. AB - BACKGROUND: A high proportion of U.S. patients with gastric carcinoma do not receive surgical treatment. To sharpen staging criteria and facilitate comparisons with surgical series, an analysis of patients whose treatment included gastrectomy was undertaken. In addition, to evaluate the "different disease" hypothesis as an explanation for superior Japanese results, outcomes for Japanese Americans were examined. METHODS: Data were obtained from National Cancer Data Base (NCDB) reports of 50,169 gastric carcinoma cases diagnosed during the years 1985-1996 and treated with gastrectomy. In addition to demographic and treatment information, 5-year and 10-year relative survival rates are presented, with stage defined according to fifth edition American Joint Committee on Cancer (AJCC) staging procedures. RESULTS: Stage-stratified 5-year and 10-year relative survival rates were as follows: Stage IA, 78%/65%; Stage IB, 58%/42%; Stage II, 34%/26%; Stage IIIA, 20%/14%; Stage IIIB, 8%/3%; and Stage IV, 7%/5%. Stage-stratified survival for Japanese Americans was higher. Males had a poorer prognosis than females, and the male-to-female ratio for Japanese Americans was lower. Proximal tumors were associated with a worse prognosis than distal tumors; the proportion of Japanese Americans with proximal disease was less than in the overall patient group. Japanese Americans underwent resection of adjacent organs less frequently. In this series, adjuvant therapy did not substantially affect survival. Overall, 20% were 10-year survivors; of these, 67% were lymph node negative and 98% had /= 15 lymph nodes analyzed. Stage migration was evident in cases with 100-fold difference in implant-retained protein dose. When rhBMP-2 and rhBMP-4 were implanted with the carriers, more rhBMP-2 was retained in an implant, and the osteoinductive potency of rhBMP-2 typically was higher than rhBMP-4 at low implantation doses. We conclude that protein pI plays a significant role in the local retention of implanted rhBMP and that higher retention yields a higher osteoinductive activity. PMID- 10679689 TI - The study of surface transformation of pulsed laser deposited hydroxyapatite coatings. AB - Hydroxyapatite (HA) coatings generally exhibit very good biocompatibility owing to their compositional resemblance to the natural hard tissue and to bioactive properties that are directly related to surface transformations in physiological fluids. In this study, two types of porous HA coatings produced with pulsed laser deposition were tested with respect to their dissolution/reprecipitation in a semidynamic simulated physiological solution. Coatings with higher porosity produced with a 355-nm wavelength laser exhibited significant reprecipitation earlier than those produced with a 266-nm wavelength laser. The dissolution of the non-HA phases played a major role in the reprecipitation of HA-like material as indicated by X-ray diffraction (XRD). The coatings' Ca/P ratio became closer to the theoretical value of HA. The newly formed HA had imperfect crystal structure and/or small crystal size as suggested by XRD. The reprecipitation resulted in a very dense morphology as shown by scanning electron microscopy, suggesting a mechanically strong structure after reprecipitation. Despite undergoing dissolution and reprecipitation, the coatings showed sufficient stability in the solution, as XRD and energy-dispersive X-ray studies indicated no significant loss of the coatings. The stability of these HA coatings and their ability to cause reprecipitation of HA in the simulated physiological solution showed the potential of these coatings for clinical applications. PMID- 10679690 TI - Structural and morphological study of pulsed laser deposited calcium phosphate bioceramic coatings: influence of deposition conditions, laser parameters, and target properties. AB - Calcium phosphate (CaP) bioceramics, especially hydroxyapatite (HA), have been used as coatings on implants owing to their biocompatible properties. The commercial practice for applying HA coating, plasma spraying, has some disadvantages which limit the long-term stability of the implants. Pulsed laser deposition (PLD) is being investigated as an alternative technique. The purpose of this research was to systematically study the effect of various parameters of the PLD process on the properties of CaP coatings. In this study, three types of HA targets and two laser wavelengths were used to make six categories of coatings. Predominantly crystalline HA coatings were produced under all six categories at optimum conditions, although small amounts of minor phases sometimes were found. Sufficient coating/substrate bond strength was also obtained. A wide variety of coating morphologies was obtained, from rather dense and uniform to rough and porous. The important factors that affected the morphology included target properties, vacuum level, deposition temperature, and laser wavelength and energy density. PLD's ability to produce both amorphous and crystalline, and both smooth/dense and rough/porous coatings may be a unique advantage. PMID- 10679691 TI - Foreign-body reaction to dermal sheep collagen in interferon-gamma-receptor knock out mice. AB - This study was performed to gain more insight into the role of interferon-gamma (IFN-gamma), a potent macrophage activator, in the foreign-body reaction to hexamethylenediisocyanate-crosslinked dermal sheep collagen (HDSC). Because the results of earlier studies aimed at modulating the foreign-body reaction in AO rats by local or systemic treatment with anti-IFN-gamma were not completely unambiguous, we extended our investigations to IFN-gamma-receptor knock-out (KO) mice. Several parameters (i.e., macrophages, giant cells, T-cells, B-cells, granulocytes, expression of MHC class II, stroma formation, and degradation and calcification of the biomaterial) were compared between wild-type (WT) and KO mice. Remarkably, the foreign-body reaction was very similar in WT and KO mice. In both, giant cells were formed, but in contrast to previous results in AO rats, phagocytosis of HDSC bundles occurred hardly at all up to 9 weeks, and MHC class II expression was minimal. Stroma formation and vascularization were high and calcification occurred. T-cells comprised less than 1%; a few plasma cells were present in the KO mice at later time points. Granulocytes, mainly eosinophils, were present at all explantation time points. Because of the similar results in WT and KO mice, we question whether IFN-gamma plays a role at all in the foreign body reaction in mice. PMID- 10679692 TI - In vitro stability of plasma-sprayed hydroxyapatite coatings on Ti-6Al-4V implants under cyclic loading. AB - The success of hydroxyapatite (HA)-coated Ti-6Al-4V implants relies on the long term stability of HA coatings. In this study, the mechanical stability of plasma sprayed HA coatings on Ti-6Al-4V implants under four-point cyclic bending was systematically investigated in both air and simulated body fluid (SBF) environments at room temperature. To have a clear view of the microscale damage evolution, the surface morphology change of HA coatings during cyclic loading was carefully examined by scanning electron microscopy at the same locations on the coating surfaces after four-point bending for 4, 6.5, 8.5, and 10 million cycles. Also, possible changes of other characteristics such as thickness, weight, crystallinity, and residual stress of HA coatings were measured as a function of loading cycles. Up to 10 million cycles of bending in air and SBF, we found no significant microcracking or coating spalling on the surface of coatings, and no significant changes in thickness, weight, crystallinity, or residual stress of the plasma-sprayed HA coatings. The experiment results indicate that thickness and crystallinity had no effects on the stability of the HA coatings. HA coating resistance to the cyclic four-point bending might result from the stress shielding effects of preexisting microcracks in the coatings. PMID- 10679694 TI - Effect of external stresses on calcium phosphate glass investigated by IR spectroscopy. PMID- 10679693 TI - Processing cell-seeded polyester scaffolds for histology. AB - Biodegradable 3-dimensional scaffolds of various morphologies are currently being developed for tissue engineering. Poly(lactide-co-glycolide)s (PLGAs) of various lactide to glycolide ratios are frequently used for such applications. Tissue engineering involves an in vitro stage during which cells are seeded onto scaffolds and allowed to settle and/or grow for various time periods. To assess cell distribution and/or tissue formation throughout the scaffolds during this in vitro stage, techniques such as confocal microscopy and magnetic resonance imaging have been applied. However, such cultured scaffolds have been refractory to histological evaluation because of numerous technical difficulties. We describe a method to prepare histological sections of cell cultured PLGA scaffolds for tissue engineering. The technique involves in situ labeling of cultured scaffolds, infiltration of the scaffolds with a 10% poly(vinyl alcohol) solution under a low vacuum, and cryosectioning of samples onto acid-treated glass coverslips. Sections obtained with this technique show cell distribution and cell-tissue morphology on the pore wall structures of entire centimeter-thick scaffolds. This rapid and easy technique allows for fast evaluation of tissues grown on biodegradable scaffolds. PMID- 10679696 TI - Effect of hemodynamic conditions on sonographic measurements of peak systolic velocity and arterial diameter in patients with peripheral arterial stenosis. AB - PURPOSE: We measured changes in peak systolic velocity ratio and sonogaphic vascular diameter during different hemodynamic conditions in patients with femoral or iliac artery stenosis. METHODS: In 35 patients with isolated femoral or iliac artery stenosis, prestenotic and intrastenotic peak systolic velocity and inner vascular diameter were calculated using color Doppler sonography and gray-scale sonography, respectively. The measurements were performed with the patient at rest (baseline), after leg exercise, and again after oral administration of 10 mg of the vasodilator nifedipine. RESULTS: The mean prestenotic and intrastenotic peak systolic velocity and the peak systolic velocity ratio (intrastenotic/prestenotic peak systolic velocity) were 70 +/- 31 cm/second, 360 +/- 130 cm/second, and 6.5 +/- 3.6 at baseline; 78 +/- 37 cm/second, 404 +/- 171 cm/second, and 6.6 +/- 4.2 after leg exercise; and 71 +/- 30 cm/second, 353 +/- 109 cm/second, and 5.9 +/- 3.2 after nifedipine administration. The mean prestenotic and intrastenotic diameter and percentage of diameter reduction were 5.9 +/- 3.2 mm, 2.3 +/- 1.1 mm, and 59 +/- 13% at baseline; 4.8 +/- 2.4 mm, 2.0 +/- 1.3 mm, and 62 +/- 13% after leg exercise; and 5.9 +/- 2.9 mm, 2.5 +/- 1.0 mm, and 54 +/- 14% after nifedipine administration. Only the difference in intrastenotic diameter after leg exercise was significantly different from baseline. CONCLUSIONS: The peak systolic velocity ratio in peripheral arterial stenosis seems to be relatively independent of the hemodynamic conditions and cannot be used for investigations of vasomotion of stenotic arterial segments during different hemodynamic conditions. PMID- 10679697 TI - Cerebrovascular reserve capacity in patients with hyperlipidemia. AB - PURPOSE: Because recent data are conflicting, it is not certain whether hyperlipidemia is an independent risk factor for cerebrovascular diseases. Decreased cerebrovascular reserve capacity refers to the decreased ability of the cerebral arterioles to adapt in critical conditions and probably predicts a higher risk of stroke. The aim of this study was to compare cerebrovascular reserve capacity in hyperlipidemic patients and healthy controls using transcranial Doppler sonography. METHODS: Thirty-four hyperlipidemic patients and 21 healthy controls were examined. With transcranial Doppler sonography, the mean blood flow velocity in the middle cerebral artery was registered at rest and at 5, 10, 15, and 20 minutes after intravenous administration of 1,000 mg acetazolamide. Cerebrovascular reactivity and reserve capacity were calculated from mean blood flow velocities. Various laboratory measurements were also made and assessed for correlation with resting cerebral blood flow velocity and cerebrovascular reserve capacity. RESULTS: No significant differences could be observed between controls and hyperlipidemic patients in cerebrovascular reactivity or cerebrovascular reserve capacity. No correlation was found between various laboratory measurements and resting cerebral blood flow velocity or cerebrovascular reserve capacity. CONCLUSIONS: We could not demonstrate any differences in cerebrovascular reserve capacity between hyperlipidemic patients and healthy controls. Thus, the vasodilatory ability of the cerebral arterioles seems to remain unchanged in this patient group and is not correlated with the severity of hyperlipidemia. PMID- 10679698 TI - Transabdominal pelvic sonography in the preoperative evaluation of patients with congenital adrenal hyperplasia. AB - PURPOSE: We assessed the clinical value of transabdominal pelvic sonography in the preoperative evaluation of patients with congenital adrenal hyperplasia (CAH) who required feminizing genitoplasty. METHODS: From 1987 to 1998, 31 patients with female pseudohermaphroditism due to CAH underwent feminizing genitoplasty. The median age of the patients was 9 months (range, 1-18 years). Radiologic evaluation performed before surgical reconstruction included retrograde genitography in the first 10 patients and sonographic examination in all 31 patients. Imaging was used to evaluate the anatomic positions and the length of the vagina, whether the junction of the vagina and the urogenital sinus occurred distal or proximal to the pelvic floor, and the presence of internal genitalia. RESULTS: Abdominal sonography identified internal female genitalia in all 31 patients, identified the anatomic shape and position of the vagina in 30 patients (97%), and confirmed the site of communication between the vagina and the urogenital sinus relative to the pelvic floor in 28 patients (90%). Sonographic findings were confirmed by intraoperative panendoscopy. Genitography was less useful than sonography, identifying the site of communication between the vagina and urogenital sinus in only 6 (60%) of 10 patients. CONCLUSIONS: In patients with CAH undergoing vaginal reconstruction, sonography provides adequate information about the anatomy of the vagina and urogenital sinus for surgical decision-making. PMID- 10679699 TI - Demonstration of gastric submucosal lesions by high-resolution transabdominal sonography. AB - PURPOSE: We evaluated the accuracy of high-resolution transabdominal sonography (TAUS) in identifying and characterizing gastric submucosal masses previously detected by endoscopy. METHODS: Patients given endoscopy for suspected submucosal gastric lesions and 2 patients with gastric wall cysts were enrolled. Patients underwent TAUS and then endoscopic sonography (EUS) on the same day, and the sonographic results were compared with endoscopic and histologic findings. RESULTS: Among 101 patients with gastric submucosal masses on endoscopy, TAUS revealed a mass in 94 (93%); of these 94 patients, 60 underwent EUS. The final diagnoses were 31 leiomyomas, 10 leiomyosarcomas, 13 ectopic pancreases, 2 cysts, and 1 glomus tumor, 1 carcinoid tumor, 1 lipoma, and 1 fibroma. Leiomyomas (mean size, 3.2 cm) and leiomyosarcomas (mean size, 7.1 cm) were shown sonographically to originate from the muscular layer. Ectopic pancreases (mean size, 1.2 cm) were shown to originate from the submucosal layer and had a homogeneously echogenic pattern. Gastric cysts were found in the submucosal layer and were anechoic. The glomus tumor and the carcinoid tumor were found in the muscular layer and were hypoechoic. The lipoma and the fibroma were located in the submucosal layer and were echogenic on TAUS. CONCLUSIONS: TAUS had a detection rate of 93% in visualizing submucosal gastric masses previously identified by endoscopy. TAUS is less invasive than EUS and can be used to follow submucosal gastric masses that are not excised. PMID- 10679700 TI - Sonographic detection of visceral adhesion in percutaneous drainage of afferent loop small-intestine obstruction. AB - To facilitate the percutaneous drainage of an afferent-loop small-intestine obstruction, we used sonography to detect visceral adhesions and select a safe puncture route. The portion of the small intestine that was fixed to the anterior abdominal wall was sonographically identified by using a high-frequency transducer to locate the area of restricted visceral sliding. The needle was then inserted into the intestine. In 3 cases, we have found that this technique improves the confidence of the physicians who perform the percutaneous drainage and may help to minimize the risks associated with the percutaneous drainage. PMID- 10679701 TI - Early prenatal sonographic diagnosis of twin triploid gestation presenting with fetal hydrops and theca-lutein ovarian cysts. AB - The presence of theca-lutein ovarian cysts in the early second trimester of pregnancy is highly suspicious for a complete hydatidiform molar pregnancy but can be seen in association with a partial mole. Theca-lutein cysts may occur following hormonal stimulation for assisted reproductive techniques or in association with multiple gestations. Rare causes include immune and nonimmune fetal hydrops, maternal hypothyroidism, and triploid gestations. We report a case of a monochorionic twin gestation in which prenatal sonography demonstrated multiple anomalies and hydrops in each twin and bilateral theca-lutein ovarian cysts. Triploidy in both twins and a partial hydatidiform mole were confirmed at pathologic examination. PMID- 10679702 TI - Sonographic detection of delayed small bowel perforations after blunt abdominal trauma. AB - Isolated perforations of the small bowel resulting from blunt abdominal trauma are rare and difficult to diagnose because of their variable presentation, especially when their appearance is delayed. We report 2 such cases that were diagnosed by sonography and emphasize the usefulness of sonography in detecting bowel injuries after blunt abdominal trauma. PMID- 10679703 TI - Color Doppler sonography of uterine arteriovenous malformation. AB - Arteriovenous malformation of the uterus is a rare but potentially life threatening lesion. We report a case of arteriovenous malformation of the uterus in a 19-year-old nulliparous woman. The diagnosis was made with color Doppler sonography and was confirmed histologically. Color Doppler sonography helps differentiate uterine arteriovenous malformation from other entities that have a similar appearance on gray-scale sonograms. PMID- 10679704 TI - Prenatal sonographic detection of a lipomeningocele as a sacral lesion. AB - We present a case of a lipomeningocele in a newborn. Prenatal sonography revealed dysraphia and a 3.8 x 4.3 cm, semisolid, echogenic mass that was continuous with the sacral area and bulged posteriorly under the skin. The mass was diagnosed after birth as a lipomeningocele based on the results of MRI. This diagnosis was confirmed histologically. PMID- 10679705 TI - Sonographic diagnosis of a small fistulous communication between a subphrenic abscess and a perforated duodenal ulcer. AB - We report a case of a fistula between a subphrenic abscess and a perforated duodenal ulcer diagnosed by sonography and confirmed by CT. The sonographic findings included a subphrenic fluid collection connected to the anterior aspect of the superior duodenum by a nonpulsatile, anechoic tubular lesion. Manual compression of the upper epigastrium resulted in movement of echogenic debris from the antrum and superior duodenum through the fistulous tract into the abscess. PMID- 10679706 TI - Type I interferons and chronic inflammatory demyelinating polyneuropathy: treatment or cause? PMID- 10679707 TI - Mechanisms of paresthesias arising from healthy axons. AB - Paresthesias are common manifestations of central and peripheral pathological processes and are due to ectopic impulse activity in cutaneous afferents or their central projections. Cutaneous afferents are more excitable than motor axons, due to differences in their biophysical properties. These differences probably include more persistent Na(+) conductance and inward rectification on cutaneous afferents, properties which probably confer greater protection from impulse dependent conduction failure but create a greater tendency to ectopic activity. Ectopic discharges can be induced in normal afferents by four maneuvers: hyperventilation, ischemia, release of ischemia, and prolonged tetanization. The alkaline shift produced by hyperventilation selectively increases the persistent Na(+) conductance, while the membrane depolarization produced by ischemia affects both transient and persistent Na(+) channels. Postischemic and posttetanic paresthesias occur when hyperpolarization by the Na(+)/K(+) pump is transiently prevented by raised extracellular K(+). The electrochemical gradient for K(+) is reversed, and inward K(+) currents trigger regenerative depolarization. These mechanisms of paresthesia generation can account for paresthesias in normal subjects and may be relevant in some peripheral nerve disorders. PMID- 10679708 TI - Electrophysiological studies of myoclonus. AB - As myoclonus is often associated with abnormally increased excitability of cortical structures, electrophysiological studies provide useful information for its diagnosis and classification and about its generator mechanisms. The EEG-EMG polygraph provides the most essential information about the myoclonus of interest. Jerk-locked back averaging and evoked potential studies combined with recording of the long latency, long loop reflexes are useful to further investigate the pathophysiology of myoclonus, especially that of cortical myoclonus. A recent advance in magnetoencephalographic techniques has contributed significantly to the elucidation of some of the cortical mechanisms underlying myoclonus. Elucidation of physiological mechanisms underlying myoclonus in each individual patient is important for selecting the most appropriate treatment of choice. PMID- 10679709 TI - Amyotrophic lateral sclerosis: early contributions of Jean-Martin Charcot. AB - Amyotrophic lateral sclerosis is historically an important entity because its manifestations involve distinct signs that can be correlated with gray and white matter lesions at specific sites within the central nervous system. Working at the end of the nineteenth century, the celebrated neurologist, Jean-Martin Charcot, used this disorder as a prototypic example of the power of his research method, termed "methode anatomoclinique." Using clinical cases and autopsy material, he showed how anatomical lesions in the nervous system could be accurately determined by the presence of carefully analyzed clinical signs. Charcot's work on amyotrophic lateral sclerosis brought together neurological entities formerly considered as disparate disorders, primary amyotrophy and primary lateral sclerosis. In addition, these studies contributed to the understanding of spinal cord and brain stem anatomy and the organization of the normal nervous system. Because of Charcot's fundamental contributions, the eponym "Charcot's disease" has been used internationally in association with amyotrophic lateral sclerosis. PMID- 10679710 TI - Nerve conduction studies in amyotrophic lateral sclerosis. AB - We studied 137 ulnar nerves and abductor digiti minimi (ADM) muscles in 70 patients with amyotrophic lateral sclerosis (ALS), and correlated the results with ADM strength graded on the Medical Research Council (MRC) scale, to address the potential value of a standardized neurophysiological assessment of this nerve muscle system. The ulnar nerves of 35 normal subjects matched for age, gender, and height served as controls. Reduced compound muscle action potential (CMAP) amplitude and area in the ADM muscle recordings correlated strongly with weakness. Distal motor latency, proximal conduction time, and F-wave frequency were abnormal with minimally detectable weakness. In weaker ADM muscles, conduction velocities and F-wave latencies were also abnormal. Conduction block was never observed and sensory potentials were normal. An "ALS neurophysiological index" was derived from these ulnar nerve studies and consisted of the expression: (CMAP amplitude/DML) x F frequency -, where F frequency was expressed as the number of F responses recorded in 20 trials. This index was strongly correlated with ADM weakness (r = 0.74, P < 0.001). Neurophysiological studies restricted to a single nerve-muscle system, the ulnar nerve/ADM, appear potentially useful in objectively assessing change in ALS. PMID- 10679711 TI - Central scalp projection of the N30 SEP source activity after median nerve stimulation. AB - Conflicting results have been reported about abnormalities of the N30 somatosensory evoked potential (SEP) in movement disorders. In these studies, the N30 amplitude was measured in the frontal scalp region. Our aim was to identify the scalp electrodes recording the genuine activity of the N30 generator. In 18 subjects, we recorded the scalp SEPs from 19 electrodes and found a negative potential around 30 ms reaching its maximal amplitude in the frontal region. However, neither simple visual inspection of the frontal traces nor topographic analysis could distinguish the N24 from the N30 component of the frontal negativity. Brain electrical source analysis of SEPs showed that a four dipolar source model could well explain the scalp SEP distribution. We calculated the scalp field distributions of the source activities as modeled from the scalp recordings and observed that the maximal field distribution reflecting the activity of the N30 source was in the central region, whereas that reflecting the N24 source activity was frontal. We conclude that the negative response recorded around 30 ms in the central traces represents "genuine" N30 source activity, whereas the frontal negativity, which is higher in amplitude, is a mixture of the activities of both the N30 and N24 sources. PMID- 10679712 TI - Quantitative EMG and motor unit recruitment threshold using a concentric needle with quadrifilar electrode. AB - According to Henneman's size principle, small motor units are recruited before large ones. We used the electromyographic (EMG) signal decomposition technique to determine the quantitative relationships between five motor unit action potential (MUAP) parameters (amplitude, duration, area, thickness, and size index) and the recruitment threshold of the motor units recruited up to 50% of the maximum voluntary contraction in the first dorsal interosseous, biceps brachii, rectus femoris, and tibialis anterior muscles of 5 healthy young men. In each muscle, the amplitude, duration, area, and size index had significant, positive high correlations with the motor unit recruitment thresholds. We conclude that the size principle applies to recordings made with concentric needle EMG electrodes under special recording conditions, and therefore that more importance should be attached to the patient's contraction force during EMG examinations in order to evaluate MUAPs for electrodiagnostic purposes. PMID- 10679713 TI - Intraoperative monitoring of the inferior alveolar nerve during mandibular sagittal-split osteotomy. AB - In order to evaluate the risk of nerve injury and to prevent iatrogenic damage at different stages of bilateral sagittal-split osteotomy (BSSO) of the mandible, we monitored the function of the inferior alveolar nerve (IAN) continuously on both sides in 13 orthognathic patients undergoing BSSO. The IAN was stimulated at the mental foramen with two monopolar needle electrodes fixed to the dental splint, and the orthodromic sensory nerve action potentials (SNAP) of the IAN were recorded with a silver-wire electrode inserted near the oval foramen on each side. The latencies, amplitudes, and sensory nerve conduction velocities at baseline, after medial opening, sawing, splitting, eventual manipulation, and fixation of the mandible were analyzed. The monitoring method functioned technically well in 25 of 26 nerves. Both the surgical technique and the duration of medial opening had conspicuous effects on the function of the IAN, which led us to modify the medial approach. When finer instruments were used for retraction and the duration of medial opening was shortened to less than 10 min, the SNAP of the IAN was always preserved at this stage. Monitoring of the IAN also prevented nerve injury during splitting and fixation. This technique for intraoperative monitoring of the IAN seems to be a feasible and promising tool for objective evaluation of intraoperative events and for prevention of nerve injury during BSSO. PMID- 10679714 TI - Evidence of inability to fully activate human limb muscle. AB - The purpose of this study was to determine whether muscle activation level estimated by twitch interpolation technique was different when an electrical stimulus was applied during a dynamic force (DF; force rising) task from that when the stimulus was applied during a static force (SF; constant force) task. Fourteen subjects performed voluntary SF and DF contractions involving isometric elbow flexion at seven voluntary force levels. At each level, the electrical stimulation was applied to the surface of the biceps brachii muscle when the force was steady (SF task) and when the force was rising (DF task). The voluntary activation level of the biceps brachii muscle during the SF maximal voluntary contraction (MVC) was 98.5% and that during the DF MVC task was significantly lower (94.5%; P < 0.05). The motoneurons and/or muscle fibers may become more excitable during the DF task so that the same stimulus can recruit those that are otherwise less excitable during the SF task. PMID- 10679715 TI - Expression of HLA-DR and its enhancing molecules in muscle fibers in polymyositis. AB - Polymyositis (PM) is an autoimmune inflammatory muscle disease of unknown cause in which cellular immunity is thought to play an important pathogenic role. Class II major histocompatibility complex (class II MHC: human leukocyte antigen (HLA) DR operates as a cofactor of antigen presentation in immunological responses. There has been a major debate over whether muscle fibers themselves synthesize and express HLA-DR molecules and play a role in antigen presentation in PM pathogenesis. In this study, we demonstrated that most muscle fibers from patients with PM synthesized and expressed HLA-DR molecules on their surface. Human leukocyte antigen-DR expression was highly specific to PM. In addition, class II transactivator (CIITA), human leukocyte antigen DM (HLA-DM), and invariant chain (Ii), which are indispensable for expression of mature HLA-DR molecules and for antigen processing and presentation, were co-expressed. One of the cytokines that could induce this expression is interferon-gamma (IFN-gamma), released by activated lymphocytes. Our results indicate that in PM muscle fibers synthesize and express HLA-DR molecules and may contribute to the inflammatory responses together with lymphocytes. PMID- 10679716 TI - Activation linearity and parallelism of the superficial quadriceps across the isometric intensity spectrum. AB - The purpose of this study was to assess neuromuscular activation of the three superficial portions of the quadriceps femoris muscles during linearly increasing isometric contraction intensities. Thirty healthy volunteers were assessed for isometric electromyographic (EMG) activity of the vastus medialis (VM), vastus lateralis (VL), and rectus femoris (RF) muscles with the knee at 60 degrees of flexion. For 5 s, subjects performed isometric contractions equivalent to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90% of the average of three maximal voluntary contractions (MVC), in random order. Full-wave rectified and integrated EMG signals over the middle 3 s of each contraction were expressed as a percentage of the activity recorded during the three averaged MVCs. One sample t tests and 95% confidence intervals were calculated at each relative torque level. A two-factor analysis of variance (muscle by intensity) with repeated measures was performed to evaluate parallel activation across the intensity levels. Activation linearity was assessed via regression analysis for each muscle. VM activation was shown to be significantly lower than expected at 20-70% MVC. VL and RF activations were significantly higher than expected at 10% MVC, and RF EMG was less than expected at 40-70% MVC. EMG of VM was shown to increase significantly more than VL and RF from 80% to 90% MVC. Significant linear and quadratic relations were also demonstrated for all three muscles. Parallel activation of the superficial quadriceps muscles occurred from low to moderate intensities, whereas convergence was noted at near maximal intensities. PMID- 10679717 TI - Multiple measures of axonal excitability: a new approach in clinical testing. AB - From measurements of nerve excitability and the changes in excitability produced by nerve impulses and conditioning currents, it is possible to infer information about the membrane potential and biophysical properties of peripheral axons. Such information cannot be obtained from conventional nerve conduction studies. This article describes a new method that enables several such measurements to be made on a motor nerve quickly and reproducibly, with minimal operator intervention. The protocol measures stimulus-response behavior using two stimulus durations (from which the distribution of strength-duration time constants can be estimated), threshold electrotonus to 100-ms polarizing currents, a current threshold relationship (indicating inward and outward rectification), and the recovery of excitability following supramaximal activation. The method was tested on 30 healthy volunteers, stimulating the median nerve at the wrist and recording from the abductor pollicis brevis. The results were comparable with previously published normal data, but the recordings took less than 10 min. The convenience and brevity of the new method make it appropriate for routine clinical use. PMID- 10679718 TI - Electrophysiological evidence for afferent nerve fibers in human ventral roots. AB - This study was designed to test the hypothesis that ventral roots in humans contain afferent nerve fibers. We made direct electrophysiological recordings of compound nerve action potentials in dorsal and ventral roots in children undergoing selective dorsal rhizotomy for spastic cerebral palsy. We stimulated the saphenous or sural nerves, which are pure sensory nerves, with electrical stimuli while systematically recording from ventral and dorsal roots from L3 to S2. In addition to the dorsal root nerve action potentials which we expected, we found smaller compound nerve action potentials, which were clearly afferent, in the ventral roots. This confirms the limited amount of experimental evidence that ventral roots do contain some afferent nerve fibers. The functional significance of these observations is not yet clear. PMID- 10679719 TI - Contractile properties of the tongue's genioglossus muscle and motor units in the rat. AB - The contractile characteristics of individual mammalian tongue muscles have rarely been investigated, in contrast to spinal cord-innervated and extraocular muscles. Therefore, whole muscle and motor unit contractile forces, plus muscle fiber types, were studied in the genioglossus, the major protrusor muscle, of the rat tongue. The muscle, exclusively composed of fast-contracting units, could be activated from rostroventral hypoglossal nucleus sites only. The following figures represent the means of the contractile measures. Whole muscle twitch tension was 7.02 g, contraction time was 14.22 ms, fusion frequency was 104 Hz, maximum tetanic tension was 37.22 g, and fatigue index was 0.72. Single motor unit twitch tension was 45. 9 mg, contraction time was 11.7 ms, fusion frequency was 94.8 Hz, maximum tetanic tension was 241.95 mg, and fatigue index was 0.68. The genioglossus muscle appeared qualitatively similar to the rat styloglossus muscle, one of the two major retractor muscles of the tongue. The delineation of motor unit contractile characteristics in tongue muscle is important in our understanding of the control of tongue movement. PMID- 10679720 TI - Proximal median neuropathy secondary to humeral neck fracture. AB - Median neuropathies proximal to the wrist are uncommon and usually result from penetrating injuries, fracture dislocation of the distal humerus, or compression by fibrous bands. A 66-year-old man suffered a comminuted fracture of the proximal humerus after a fall. Electrodiagnostic studies revealed a severe proximal median neuropathy and a mild distal radial mononeuropathy. Proximal median neuropathy rarely occurs in humeral neck fracture, mostly because the median nerve is not in close contact with the humerus proximally. PMID- 10679721 TI - Serum type IV collagen increases with duration of amyotrophic lateral sclerosis. AB - We analyzed the serum levels of type IV collagen, a major component of the basement membrane, during various stages of sporadic amyotrophic lateral sclerosis (ALS). Serum concentrations of type IV collagen increased significantly with the duration of illness. Findings indicate that the metabolism of type IV collagen and the basement membrane may be affected in the disease process of ALS. PMID- 10679722 TI - Chronic inflammatory demyelinating polyneuropathy after treatment with interferon alpha. AB - Treatment with interferon-alpha (IFN-alpha) has been associated with the occurrence of a number of autoimmune disorders. We report a case of chronic inflammatory demyelinating polyneuropathy (CIDP) occurring in a patient with a chronic viral hepatitis C infection who received a novel, long-acting form of IFN alpha. After withdrawal of the interferon treatment, this patient responded to a single extended course of plasma exchange that resulted in a complete clinical remission of symptoms without relapse. PMID- 10679723 TI - Kaposi's sarcoma herpesvirus-associated Castleman's disease with POEMS syndrome. AB - The pathogenic link between POEMS (polyneuropathy, organomegaly, endocrinopathy, M protein, skin changes, and various other clinical signs) syndrome and Castleman's disease is unclear. Roles for M protein in POEMS syndrome and cytokines in systemic manifestations of multicentric Castleman's disease have been suggested. Recently, pathogenic roles of cytokines in POEMS syndrome and Kaposi's sarcoma-associated herpesvirus (KSHV) in Castleman's disease have been reported. We report on a patient with KSHV-associated Castleman's disease with POEMS syndrome, and suggest a possible role of KSHV in initiating and linking these two diseases. PMID- 10679724 TI - Simultaneous exacerbation and remission of central and peripheral demyelination. PMID- 10679725 TI - AAEM news and comments. PMID- 10679726 TI - Noninvasive determination of genome size and ploidy level in fishes by flow cytometry: detection of triploid Poecilia formosa. AB - BACKGROUND: In order to understand the evolutionary significance of single triploids among the mostly diploid Poecilia formosa we have developed a simple, noninvasive technique for DNA content and ploidy determination. METHODS: From dorsal fin clips of 14 different fish species single cell suspensions were obtained by chopping the material in 2.1% citric acid/0.5% Tween20, passing it through a 0. 6-gauge needle and incubating it for 20 min at room temperature (RT) with gentle agitation. After overnight fixation in 70% ethanol, the cells were treated with 1ml 0.5% pepsin/0.1 M HCl for 15 min at RT before adding DAPI to a final volume of 2 ml. The cells were stained for 1-3 h and then analyzed by flow cytometry. RESULTS: We obtained good measurements with CVs ranging from 1.23% to 3.36%. The poeciliid species measured contain from 1.6 to 2.0 pg/nucleus, Oryzias latipes (Medaka) exhibits a nuclear DNA content of 2.2 pg, Danio rerio (zebrafish) 4.6 pg, Tetraodon fluviatilis (freshwater fugu) 0.70 pg. All values except zebrafish are in good agreement with the literature. CONCLUSIONS: The identification of living specimens of different ploidy for breeding experiments, behavioral studies and tissue transplantations is now made possible. With slight modifications the method can be extended to a field technique, providing therefore a useful tool for a variety of researchers. PMID- 10679727 TI - Four-color multiparameter DNA flow cytometric method to study phenotypic intratumor heterogeneity in cervical cancer. AB - BACKGROUND: Multiparameter DNA flow cytometry using a one-laser bench-top flow cytometer has been restricted to three different colors. The two laser FACSCalibur has recently been introduced, allowing four-color analysis. Therefore, we optimized and extended our three-color method (Corver et al., 1994, Corver et al. 1996) to a four-color analysis of phenotypic intra-tumor heterogeneity using a bench-top flow cytometer. METHODS: First, the effect of a range of different propidium iodide (PI) and TO-PRO-3 iodide (TP3) concentrations on the coefficient of variation (CV) of the DNA histograms was measured using paraformaldehyde-fixed lysolecithin-permeabilized peripheral blood lymphocytes (PBLs) and SiHa and HeLa cervical cancer cells. Second, labeling freshly isolated cervical cancers from solid tumors was optimized with a mixture of anti-keratin antibodies. Third, the FACSCalibur hardware was modified, thereby allowing the simultaneous measurement of allophycocyanin (APC) fluorescence (FL4) in combination with FL3 pulse processing (FL3-W vs. FL3-A). The optimized procedure was then applied to cell suspensions from four different human cervical cancers to study phenotypic intratumor heterogeneity. Cell suspensions were simultaneously stained for DNA (PI, fluorescence) and three cellular antigens: (a) the epithelial cell-adhesion molecule (Ep-CAM; APC fluorescence), (b) keratin (R-phycoerythrin [RPE] fluorescence) to identify the epithelial fraction, and (c) vimentin (fluorescein-isothiocyanate [FITC] fluorescence) to label stromal cells. RESULTS: Overall, PI produced better CVs than did TP3. The optimal concentration of PI was 50-100 microM for all cells tested. Average CVs were 1.76% (PBL), 3.16% (HeLa), and 2.50% (SiHa). Optimal TP3 concentrations were 0.25-2.0 microM. Average CVs were 2. 58% (PBL), 5.16% (HeLa), and 3.96% (SiHa). Inter- or intra DNA stem line heterogeneity of Ep-CAM expression was observed in the keratin positive fractions. Vimentin-positive, keratin-negative cells were restricted to the DNA diploid fraction. CONCLUSIONS: PI is a superior DNA stain to TP3 when using intact normal PBL and human cancer cells. Four-color high-resolution multiparameter DNA flow cytometry allows the identification of intratumor subpopulations using PI as DNA stain and FITC, RPE, and APC as reporter molecules. The FACSCalibur bench-top flow cytometer can be used for this purpose, allowing the application of this technique in clinical laboratories. PMID- 10679728 TI - Detection of cell cycle subcompartments by flow cytometric estimation of DNA-RNA content in combination with dual-color immunofluorescence. AB - BACKGROUND: Correlated flow cytometric measurements of phenotype and DNA-RNA content offer detailed information on cell cycle status of subpopulations in heterogeneous cell preparations in response to stimulation. We have developed a method for flow cytometric analysis of DNA-RNA content that has been optimized for simultaneous measurement of dual-color immunofluorescence. METHODS: Nucleic acid staining was performed at low pH in the presence of saponin. DNA was stained with 7-aminoactinomycin D (7-AAD) and RNA with pyronin Y(G) (PY); both dyes were used at low concentrations, and 7-AAD was exchanged with nonfluorescent actinomycin D after DNA staining to minimize fluorochrome-fluorochrome interactions. For cell surface antigen staining, allophycocyanin was combined with pH-independent Alexa488 instead of fluorescein-isothiocyanate (FITC) because FITC is pH sensitive. RESULTS: This method identified cell cycle subcompartments in CEM cells comparable to published results on cell lines using other dyes and staining methods. Measurement of DNA-RNA content in CD8 lymphocyte subsets of human peripheral blood mononuclear cells costimulated with CD3/CD28.2 showed that, after 48 h of stimulation, 80% of CD8(+) T cells were in the proliferative state, whereas 86% of CD8(+) non-T cells remained in G(0). CONCLUSIONS: This technique permits the clear identification of cellular subpopulations by phenotype and assessment of their cell cycle status. PMID- 10679729 TI - Different expression profiles of human cyclin B1 in normal PHA-stimulated T lymphocytes and leukemic T cells. AB - BACKGROUND: In a previous work, we demonstrated with flow cytometry (FCM) methods that accumulation of human cyclin B1 in leukemic cell lines begins during the G(1) phase of the cell cycle (Viallard et al. , Exp Cell Res 247:208-219, 1999). In the present study, FCM was used to compare the localization and the kinetic patterns of cyclin B1 expression in Jurkat leukemia cell line and phytohemagglutinin (PHA)-stimulated normal T lymphocytes. METHODS: Cell synchronization was performed in G(1) with sodium n-butyrate, at the G(1)/S transition with thymidine and at mitosis with colchicine. Cells (leukemic cell line Jurkat or PHA-stimulated human T-lymphocytes) were stained for DNA and cyclin B1 and analyzed by FCM. Western blotting was used to confirm certain results. RESULTS: Under asynchronous growing conditions and for both cell populations, cyclin B1 expression was essentially restricted to the G(2)/M transition, reaching its maximal level at mitosis. When the cells were synchronized at the G(1)/S boundary by thymidine or inside the G(1) phase by sodium n-butyrate, Jurkat cells accumulated cyclin B1 in both situations, whereas T lymphocytes expressed cyclin B1 only during the thymidine block. The cyclin B1 fluorescence kinetics of PHA-stimulated T lymphocytes was strictly similar when considering T lymphocytes blocked at the G(1)/S phase transition by thymidine and in exponentially growing conditions. These FCM results were confirmed by Western blotting. The detection of cyclin B1 by Western blot in cells sorted in the G(1) phase of the cell cycle showed that cyclin B1 was present in the G(1) phase in leukemic T cells but not in normal T lymphocytes. Cyclin B1 degradation was effective at mitosis, thus ruling out a defective cyclin B1 proteolysis. CONCLUSIONS: We found that the leukemic T cells behaved quite differently from the untransformed T lymphocytes. Our data support the notion that human cyclin B1 is present in the G(1) phase of the cell cycle in leukemic T cells but not in normal T lymphocytes. Therefore, the restriction point from which cyclin B1 can be detected is different in the two models studied. We hypothesize that after passage through a restriction point differing in T lymphocytes and in leukemic cells, the rate of cyclin B1 synthesis becomes constant in the S and G(2)/M phases and independent from the DNA replication cycle. PMID- 10679730 TI - Enrichment for submitotic cell populations using flow cytometry. AB - BACKGROUND: One of the most dramatic events during the course of the mammalian cell cycle is mitosis, when chromosomes condense and segregate, the nuclear envelope breaks down, and the cell divides into two daughter cells. Although cells undergoing mitosis are cytologically distinguishable from nonmitotic cells, few molecular markers are available to specifically identify mitotic cells, especially cells within different stages of mitosis. METHODS: We applied the flow cytometric method of Juan et al. (Cytometry 32:71-77, 1998) to obtain cells with various levels of the molecular markers cyclin B1 and phosphorylated histone H3; fluorescence microscopy was then used to identify sorted cells in different stages of mitosis. RESULTS: We observed the substantial enrichment of submitotic cell populations. CONCLUSIONS: This method represents an effective approach to obtain an enriched population of submitotic cells without the use of drug treatments or prior synchronization. PMID- 10679731 TI - Multiplexed single nucleotide polymorphism genotyping by oligonucleotide ligation and flow cytometry. AB - BACKGROUND: We have developed a rapid, high throughput method for single nucleotide polymorphism (SNP) genotyping that employs an oligonucleotide ligation assay (OLA) and flow cytometric analysis of fluorescent microspheres. METHODS: A fluoresceinated oligonucleotide reporter sequence is added to a "capture" probe by OLA. Capture probes are designed to hybridize both to genomic "targets" amplified by polymerase chain reaction and to a separate complementary DNA sequence that has been coupled to a microsphere. These sequences on the capture probes are called "ZipCodes". The OLA-modified capture probes are hybridized to ZipCode complement-coupled microspheres. The use of microspheres with different ratios of red and orange fluorescence makes a multiplexed format possible where many SNPs may be analyzed in a single tube. Flow cytometric analysis of the microspheres simultaneously identifies both the microsphere type and the fluorescent green signal associated with the SNP genotype. RESULTS: Application of this methodology is demonstrated by the multiplexed genotyping of seven CEPH DNA samples for nine SNP markers located near the ApoE locus on chromosome 19. The microsphere-based SNP analysis agreed with genotyping by sequencing in all cases. CONCLUSIONS: Multiplexed SNP genotyping by OLA with flow cytometric analysis of fluorescent microspheres is an accurate and rapid method for the analysis of SNPs. PMID- 10679732 TI - Apoptosis can be detected in attached colonic adenocarcinoma HT29 cells using annexin V binding, but not by TUNEL assay or sub-G0 DNA content. AB - BACKGROUND: Induction of apoptosis in adherent cell lines is associated with cell loss from the substratum. In this study the adenocarcinoma cell line, HT29, treated with indomethacin (400microM) has been employed as a model system to demonstrate how flow cytometric analysis can be used to quantify the changes that occur during this process. METHODS: Adherent and floating cell populations have been analyzed independently for effects on cell number, cell cycle characteristics and apoptosis using TUNEL assay and Annexin V binding. In addition apoptosis has been assessed using DNA laddering and morphology. RESULTS: Apoptosis was detected in adherent cells treated with indomethacin using Annexin V binding but not by other techniques employed in this study. In contrast, analysis of "floating" cells revealed the presence of apoptotic cells both in control and indomethacin treated cells using all the techniques employed. However quantification by flow cytometry showed that a significantly higher proportion of control "floaters" were late apoptotic/necrotic rather than apoptotic. DISCUSSION: The data here illustrate the need to interpret measures of apoptosis in adherent cell lines with care and the value of using flow cytometric techniques in the quantitative evaluation of the process. PMID- 10679733 TI - Simultaneous analysis of radio-induced membrane alteration and cell viability by flow cytometry. AB - BACKGROUND: Modifications of intracellular transfer, resulting from a loss of membrane integrity may contribute toward setting the cell onto the pathway of apoptosis. METHODS: We have developed an original technique of measuring simultaneously, with flow cytometry, changes in membrane fluidity and cell death status. Our aim was to assess the extent to which radio-induced cell death and membrane alterations are linked. Investigations were performed on lymphocytes 24 h after whole human blood gamma-irradiation. RESULTS: Our results confirmed the expected increase in the percentage of apoptotic cells as a function of dose, but revealed that the percentage of necrotic cells appeared stable after irradiation. At the same time, the fluorescence anisotropy of the living lymphocyte subpopulation decreased significantly and dose dependently as measured 24 h post irradiation. With TMA-DPH, the anisotropy index of apoptotic lymphocytes was always lower than that of the viable lymphocyte subpopulation. On the other hand, 1,6-diphenyl-1,3,5-hexatriene (DPH) anisotropy was similar in apoptotic and viable cells after irradiation. These findings suggest that apoptotic lymphocytes are characterised by a membrane fluidization that mainly occurs on the cell membrane surface. CONCLUSION: Our study made technical advances in using cytometric fluorescence anisotropy measurement as an early biological indicator of apoptosis after cellular exposure to ionising radiation. PMID- 10679734 TI - Flow cytometry analysis of atherosclerotic plaque cells from human carotids: a validation study. AB - BACKGROUND: Atherosclerotic plaques are heterogeneous vascular lesions. Changes in cell plaque composition are fundamental events inside the plaque microenvironment that are strictly related to the clinical outcome of these lesions (organ damage). The knowledge of these modifications may help to better understand the pathophysiological mechanisms of atherosclerosis. METHODS: We report on a flow cytometry method to characterize and quantify the cell subpopulations in human atherosclerotic plaques. Cells were obtained from endarterectomy specimens after collagenase digestion. Both surface and intracytoplasmic antigens were labeled. RESULTS: Our data demonstrated that the method we described allowed the characterization of cell populations that compose the atherosclerotic plaque, avoiding contamination by tunica media smooth muscle cells and the noise of cellular debris. Moreover this validation study showed that about 50% of cells in the atherosclerotic plaques are inflammatory mononuclear cells (T lymphocytes and monocytes/macrophages). CONCLUSIONS: Reproducible quantitative methods for cell population characterization may increase the understanding of pathophysiological mechanisms responsible for plaque progression. The methodology herein described gave us the possibility of quickly calculating the relative amount of each cell population and studying both surface and intracellular markers to analyze the functional stage of the cells. The clinical correlation was not assessed in the present study, because we used a small patient group to validate the method, but should be the subject of further analyses in a larger patient population. PMID- 10679735 TI - Determination of liposome size distribution by flow cytometry. AB - BACKGROUND: An essential parameter that describes the quality of liposome suspensions is the mean size, respectively the size distribution. Currently several analytical methods including laser light scattering techniques (LLST) are being employed. METHODS: Here we present an alternative technique using flow cytometry (FCM) to characterize uni- and polydisperse suspensions. As model liposomes preparations containing dipalmitoylphosphatidylcholine (DPPC) were used. A constant number of particles (1,500/s) in the fluid stream and a representative number of 10,000 particles of each sample was measured. Fluorescence-labeled latex beads were measured identically, and their side scatter signals were calibrated and correlated to the results obtained with liposome vesicles. RESULTS: Evaluation of the measurement and validation of the FCM results in comparison to LLST confirm the reliability of results obtained with our method. Latex beads in the range of 100-1000 nm were used for calibration to classify liposomes. Although measurement characteristics and calculation in both methods are basically different, very good agreement of the results was achieved. CONCLUSIONS: Demonstration of stability, reproducibility, and reliability of results make the employment of this method acceptable for an adequate routine analysis technique. PMID- 10679736 TI - Analyses of quality assessment studies using CD45 for gating lymphocytes for CD3(+)4(+)%. AB - BACKGROUND: The purpose of this study was to assess whether laboratories which do not use CD45 for gating lymphocytes with three- (or four-) color flow cytometry (non-CD45 laboratories) for CD3(+)4(+)% and CD3(+)8(+)% do worse on quality assessment (QA) studies than laboratories which do use CD45 (CD45 laboratories). METHODS: Data came from blood specimens donated by 62 donors (50 HIV-positive) assayed over 2 years (November, 1996-October, 1998) by 35 laboratories in the NIAID DAIDS Flow Cytometry QA Program. RESULTS: Non-CD45 laboratories were significantly more likely to be classified as having unacceptable inter laboratory results (far from the group median) than CD45 laboratories (5.6% vs 1.5%, P = 0.005 for CD3(+)4(+)%; 10.4% vs 5.0%, P = 0.007 for CD3(+)8(+)%). The intra-laboratory range of results on blinded replicates was significantly more likely to be deemed unacceptable (range >4%) in non-CD45 laboratories than in CD45 laboratories for CD3(+)8(+)% (14. 5% vs 3.5%, P = 0.002) but not for CD3(+)4(+)% (2.6% vs 1.5%, P = 0. 62). These differences in favor of CD45 gating were observed even though the non-CD45 laboratories had been doing three-color flow cytometry in the QA program significantly longer (P = 0.05) than the CD45 laboratories, and so would be expected to have fewer problems with the assay. CONCLUSIONS: Laboratories which choose to use a single CD3/CD4/CD8 tube for immunophenotyping may be sacrificing both accuracy and reproducibility. PMID- 10679737 TI - Role of immunological lymphocyte subset typing as a screening method for lymphoid malignancies in daily routine practice. AB - BACKGROUND: The major diagnostic role of peripheral lymphocyte subset typing is to distinguish between malignant and reactive conditions. METHODS: The present study evaluates the screening efficacy of flow cytometric lymphocyte subset typing for the presence of a lymphoid malignancy. Four hundred samples were analyzed with a combination of anti-T-, B-, and natural killer (NK)-cell monoclonal antibodies. RESULTS: Two hundred and twenty (55%) samples showed a normal distribution of lymphocyte subsets, 73 (18%) samples exhibited unspecific alterations of lymphocyte subsets, 19 (5%) samples exhibited a reactive phenotype typical of Epstein-Barr virus/cytomegalovirus (EBV/CMV) infection, and 88 (22%) samples expressed a phenotype suggestive of lymphoma. The most predictive independent factor of a lymphoma-specific phenotype was the absolute lymphocyte count (P = 0.0001, odds ratio 73.225). Seventy-eight percent of samples containing >/=4 x 10(9)/l lymphocytes and 2% of samples with lymphocyte counts <4 x 10(9)/l exhibited a lymphoma-specific phenotype. The specificity of the referring clinical comment was the second best predictor of a lymphoma-specific typing outcome (P = 0.0001, odds ratio 19.589). The independent predictive values of lymphocyte morphology and of relative lymphocyte counts were of borderline significance. CONCLUSIONS: The use of flow cytometric lymphocyte subset typing as a diagnostic screening method for lymphoma should be restricted to cases of unexplained elevation of absolute lymphocyte counts with or without morphological atypias and to cases with definite clinical symptoms of lymphoma. PMID- 10679738 TI - Lymphocyte subsets and specific T-cell immune response in thalassemia. AB - Infection is very common in thalassemia and is one of the major causes of death. To date, it is not quite clear why these patients are susceptible to infection. In this study, lymphocyte immunophenotyping for CD3(+) (T-cells), CD3(+)CD4(+) (T helper/inducer cells), CD3(+)CD8(+) (T-suppressor/cytotoxic cells), CD3(-)CD19(+) (B-cells), and CD3(-)CD16/56(+) (natural killer cells) subsets and expression of the activation antigen CD69 on CD3(+)CD4(+) and CD3(+)CD8(+) T-cells were determined in the whole blood of thalassemia patients, using a three-color flow cytometric technique. Results showed that only splenectomized beta thalassemia/hemoglobin (Hb) E patients displayed a marked increase in absolute number of all lymphocytes. In addition, splenectomized beta-thalassemia/Hb E showed a significantly lower percentage of CD3(+) cells, with a corresponding increase in CD19(+) cells. These differences, when compared with normal subjects and other thalassemia patients, may be attributed to splenectomy. alpha thalassemia patients, on the other hand, showed no significant difference from the normal group. While lymphocyte subsets in splenectomized beta-thalassemia/Hb E patients showed an abnormal distribution, T-cell activation in these patients was not different from the activation seen in normal subjects. This implies that thalassemia patients, during the steady state of disease, appear to have normal T lymphocyte function with only moderate abnormalities of T- and B-lymphocyte subsets. PMID- 10679739 TI - Improving accuracy in the grading of renal cell carcinoma by combining the quantitative description of chromatin pattern with the quantitative determination of cell kinetic parameters. AB - The determination of grade and stage in renal cell carcinomas (RCCs) often fails to predict the actual clinical outcome for individual patients. The aim of the present work was to investigate whether it is possible to significantly improve the prognostic accuracy of the grading system by using the combination of two independent computer-assisted microscopy techniques. The first technique relates to the quantitative description of morphonuclear and nuclear DNA content features by means of the image analysis of Feulgen-stained cell nuclei, and the second quantitatively characterizes tumor growth by means of different cell kinetic parameters. These parameters consist of a duplication of a time-related parameter determined by means of the technique of using silver-stained proteins in interphase nucleolar organizer regions (AgNOR), a proliferation index determined by means of MIB-1 immunohistochemistry, and an apoptotic index determined by means of the terminal dUTP nick end labeling technique. The prognostic value of these quantitative features was investigated in a series of 60 RCCs. The quantitative analysis of Feulgen-stained nuclei made it possible to identify subgroups of patients with significantly different prognoses in both grade II and grade III RCCs. We labeled the RCCs associated with the most favorable prognoses as grade II- and III- and those with the least favorable ones as grade II+ and III+. The two most important kinetic variables to identify patients with different clinical outcomes were the MIB-1 index and the mean AgNOR area in the MIB-1-positive cells. Three significantly different survival curves were obtained for the 53 grade II and III RCC patients. Our results show that conventional RCC grading can be significantly improved by the quantitative analysis of Feulgen stained nuclei, by cell kinetic parameter determination, and, more importantly, by combining the proliferation index with the mean AgNOR area parameter. PMID- 10679740 TI - Flow cytometric DNA ploidy, p53, PCNA, and c-erbB-2 protein expressions as predictors of survival in surgically resected gastric cancer patients. AB - In order to determine retrospectively the impact of some cytometric and immunohistochemical parameters on the overall survival of gastric cancer patients treated with surgery alone, paraffin-embedded tumor samples from 137 gastric carcinoma patients undergoing curative resection from 1987-1993 were analyzed by flow cytometry (FCM) and immunohistochemistry (p53, c-erbB-2, and PCNA expression). FCM-derived parameters were DNA ploidy and fraction of S-phase cells (SPF). Multiple regression analysis was applied to determine the prognostic significance of the conventional clinicopathologic findings together with the flow cytometric and immunohistochemical parameters on overall survival. When all parameters were entered simultaneously into the Cox regression model, stage and DNA ploidy (DNA index >1.35) clearly emerged as the only independent prognostic factors. When the stages were analysed separately, the independent prognostic factors resulted DNA ploidy in early stages (I-II) and grading in stage IIIA tumors. For stage IIIB tumors, no independent prognostic factor was found. These results indicate that the DNA ploidy pattern is a valuable predictor of survival in curatively resected gastric cancer patients, especially when less advanced tumors are taken into consideration. PMID- 10679741 TI - Variation of flow cytometric DNA measurement in 1,485 primary breast carcinomas according to guidelines for DNA histogram interpretation. AB - From 1990-1996, 1,485 previously untreated invasive breast carcinomas were sampled by a pathologist for flow cytometric DNA analysis. The aim of the present work was to study the variations of flow cytometric DNA ploidy and S-phase evaluation according to the conditions of DNA histogram interpretation. Results obtained with the American Consensus guidelines of 1993 and the Francois Baclesse Department of Pathology's own guidelines are presented. According to the percentage of events taken into account to identify a DNA aneuploid peak, the proportion of DNA diploid cases can change from 35-39%. For S-phase evaluation, although the two guidelines were quite different, the results of S-phase cutoff were identical. Whichever guidelines were used, there was a strong relationship between DNA ploidy and/or S-phase and classical clinicopathological factors (T, N, histological type, grade, receptor status, or lymphatic invasion), with the exception of age, whose correlation was discrepant with S phase according to the set of guidelines. Whichever guidelines were used, ploidy and S phase correlated strongly with survival (overall, metastasis-free, or recurrence-free). Hence we recommend the use of the American consensus guidelines, despite minor imperfections, because they are now well-known, allow a high yield in the ratio of assessable S phases, and permit standardization in the technical processing and reporting of S phases, thanks to the use of terciles. PMID- 10679742 TI - Novel functional multiparameter flow cytometric assay to characterize proliferation in skin. AB - Keratins are a group of cytoskeletal proteins that are found in human epidermis and other stratified squamous epithelia. Several different types of keratins have been described. Keratin 10 (K10) is a keratin that is expressed in well differentiated, suprabasal keratinocytes, and keratin 6 (K6) is a keratin which is associated with hyperproliferation. Psoriasis is a chronic inflammatory skin disease, and besides inflammation, disturbed differentiation and hyperproliferation are its hallmarks. In order to study the hyperproliferation associated keratinization in both well differentiated and poorly differentiated keratinocytes, and in order to assess the proliferative activity of all K10 and K6 subpopulations, simultaneous assessment of K6, K10, and DNA content is required. So far, a triple staining protocol had not been available. In the present study, we established a novel protocol for simultaneous measurement of K6, K10, and DNA content, which enables the characterization of the proliferative activity of several cellular subpopulations in epidermis. From 16 patients with psoriasis and from 15 healthy volunteers, punch biopsies were obtained. After preparation of single cell suspensions, cells were stained with the anti-keratin 10 IgG(1)-isotype monoclonal antibody RKSE60, with the anti-keratin 6 IgG(2a) isotype monoclonal antibody LHK6B, and with the DNA fluorochrome TO-PRO-3 iodide. Isotype specific secondary antibodies conjugated with phycoerythrein (PE) and fluorescein isothiocyanate (FITC) were used as the second step in the staining procedure. Controls were measured omitting the primary antibodies, and gates were set in order to differentiate between the K10 and K6 subpopulations. Samples from both psoriatic patients and healthy volunteers were than measured. Owing to the IgG specificity of RKSE60 and LHK6B, no cross-reactivity was observed between these antibodies. The triple staining with RKSE60, LHK6B, and TO-PRO-3 iodide showed subpopulations of K10 expressing cells, K10/K6 co-expressing cells, and K6 only expressing cells. There was a significant difference in the proportion of K6 expression and K10/K6 co-expression between psoriatic and normal skin. Moreover, the proliferative activity of these subpopulations could be quantified by this protocol. We concluded that a triple staining protocol for the assessment of K6, K10, and DNA content, using the monoclonal antibodies LHK6B, RKSE60, and TO-PRO-3 iodide, supplies reliable and reproducible data for cellular studies on these keratins and for studying the proliferative activity of the subpopulations of these keratins in epidermis. Moreover, the present study showed that with respect to the proportion of K6, significant differences are present between psoriatic and healthy human skin. PMID- 10679743 TI - Response of chemosensitive and chemoresistant leukemic cell lines to drug therapy: simultaneous assessment of proliferation, apoptosis, and necrosis. AB - BACKGROUND: The balance between cell proliferation and drug-induced cell death by apoptosis or necrosis plays a major role in determining response to chemotherapy. Commonly-used DNA analysis methods cannot study both parameters simultaneously. A new approach described here combines a green fluorescent membrane-intercalating dye (PKH67) with Hoechst 33342 or annexin V and propidium iodide, to allow simultaneous assessment of cell division, cell cycle status, apoptosis, and necrosis, respectively. METHODS: To test this approach, we used cultured K562 leukemic cell lines which are drug-sensitive (K562S) or drug-resistant (K562R) by virtue of whether they lack or exhibit expression, respectively, of the gp-170 (PGP) glycoprotein pump involved in multidrug resistance. RESULTS: We found that: 1) PKH67 fluorescence intensity decreases proportionately to number of cell divisions, 2) labeling with PKH67 does not alter either cell cycle distribution, as assessed by vital DNA staining with Hoechst 33342, or cell growth, and 3) using a simple threshold analysis method suitable for real-time sorting decisions, subpopulations of proliferating cells present at initial levels of >/= 10% can readily be detected after two cell division times, based on decreased PKH67 intensity. Finally, we demonstrated that after treatment of an admixture of K562S and K562R with vincristine, triple-labeling with PKH67, annexin V, and propidium iodide can be used to identify and sort those cells which remain not only viable (nonnecrotic, nonapoptotic) but actively dividing (decreased PKH67 intensity) in the presence of drug. CONCLUSIONS: Although the studies described here were carried out in a model system using cells having known drug resistance phenotypes, we expect that the methods described will be useful in ex vivo studies of clinical leukemic specimens designed to identify the role played by specific chemoresistance proteins and mechanisms in therapeutic outcomes for individual patients. PMID- 10679744 TI - Bcl-2: bax and bcl-2: Bcl-x ratios by image cytometric quantitation of immunohistochemical expression in ovarian carcinoma: correlation with prognosis. AB - Bcl-2 is a proto-oncogene which is involved in prolonging cell survival by inhibiting programmed cell death. Bax and bcl-x are members of the bcl-2 family; when overexpressed, they can counteract the ability of bcl-2 to inhibit apoptosis. This suggests a model in which the ratios of bcl-2 to bax and bcl-x can be used to determine response to therapy and prognosis. The expression of bcl 2, bax and bcl-x was studied in 50 ovarian carcinomas. The percentage of positive area immunostained (PPA) in the nucleus and cytoplasm of each ovarian carcinoma was quantitated in 15 high power fields by image cytometry. The ratios were obtained by dividing the PPA of bcl-2 by the PPA of bax and bcl-x. 17 of 50 ovarian carcinomas (34%) stained positively for bcl-2, 39 for bax (78%) and 47 for bcl-x (94%). Although there is no significant statistical correlation between expression of bcl-2, bax or bcl-x and grade (P = 0.15; P = 0. 47; P = 0.56), stage (P = 0.71; P = 0.6; P = 0.42), and overall or disease-free survival (P = 0.26; P = 0.55; P = 0.16), increased bcl-2 expression was demonstrated in patients with shortened overall and disease-free survival. Also, increased expression of bax and bcl-x was associated with increased overall and disease free survival. Bcl-2:bax and bcl-2:bcl-x ratios less than 1 are associated with survival advantage, although not statistically significant (P = 0.83; P = 0.93). Image cytometric measurement of bcl-2, bax, and bcl-x expression is feasible. There is a tendency for their expression to correlate with prognosis in ovarian carcinomas. PMID- 10679745 TI - Detection of apoptotic T lymphocytes in peripheral blood of human immunodeficiency virus (HIV)-infected subjects by apostain. AB - Apoptosis has been indicated as a mechanism of T cell depletion in HIV-infected subjects and useful in monitoring disease progression. We investigated for the presence of apoptotic T lymphocytes in 130 HIV subjects in various stages of disease by the newly developed cell permeant DNA dye Apostain. Blood was collected in EDTA, lysed in buffered ammonium chloride, fixed in freshly prepared 1% paraformaldehyde and stored in aliquots at -80 degrees C. Samples were thawed and double stained with FITC conjugated-CD3 monoclonal antibody and Apostain. Flow cytometry was then performed and T cells gated on a CD3 versus side scatter dot plot. Normal samples treated in the same manner served to establish the boundary separating non-apoptotic from apoptotic cells. There was no statistically significant association between the proportion of subjects with detectable apoptotic cells and CDC clinical categories A, B and C at the time of admission to the study, although a trend toward a lower apoptotic rate in category A (A= 29%, B=40% and C=41%) was noticed. Conversely, CDC T cell categories 2 and 3 contained significantly higher proportions of Apostain positive patients (1=6%, 2=32% and 3=49%, P=0.072, by chi(2) test). Most importantly, Apostain test identified subjects at risk of disease progression during a 3.5-7 months follow-up in CDC category B and 2 (P=0.008 and P=0.0003, by Fisher's exact test, respectively). A similar, albeit not statistically significant trend was observed also in the other categories. Not requiring extensive manipulation of fresh samples nor cumbersome culture techniques, Apostain test appears suitable for identifying HIV subjects at higher risk of disease progression in clinical settings. PMID- 10679746 TI - Comparison of DiOC(6)(3) uptake and annexin V labeling for quantification of apoptosis in leukemia cells and non-malignant T lymphocytes from children. AB - Early during apoptosis, there is a reduction in mitochondrial transmembrane potential (MTP) and externalization of phosphatidylserine (PS) in cell membrane prior to eventual cell death. Flow cytometric detection techniques targeting these changes, reduction of DiOC(6)(3) uptake upon the collapse of MTP and annexin V binding to PS have been successfully used to detect apoptotic cells. These methods have given comparable results when cell lines were used. We compared the two different techniques, DiOC(6)(3) uptake and Annexin V-propidium iodide co-labeling in the quantification of cytarabine, vincristine and daunorubicin induced apoptosis on three leukemia cell lines (HL-60, CEM, U937), and bone marrow blasts from 26 children with acute myeloid leukemia, 14 with T cell acute lymphoblastic leukemia. Anti-Fas-induced apoptosis in culture-grown peripheral blood T lymphocytes on 18 samples from 9 children with non-malignant conditions were also studied by these techniques. Our results showed that there is a correlation (P < 0. 05) between the apoptosis rates measured by these two techniques for drug-induced apoptosis in myeloid and lymphoid blasts, and for anti-Fas mAb-induced apoptosis in T lymphocytes. This data suggests that reduction of the MTP and PS externalization may be common to many apoptotic pathways and techniques targeting either of these changes may be used in quantification of apoptosis in different clinical samples. PMID- 10679748 TI - Forum: journal club PMID- 10679747 TI - CD4 count/viral load in HIV. PMID- 10679749 TI - RAPD library fingerprinting of bacterial and human DNA: applications in mutation detection. AB - Random amplified polymorphic DNA (RAPD) fingerprinting is a modification of the polymerase chain reaction (PCR), which utilises a single, arbitrarily-chosen primer to amplify a number of fragments from a given template DNA to generate a discrete "fingerprint" when resolved by gel electrophoresis. Alterations by as little as a single base in the primer sequence lead to marked alterations in the fingerprints generated with a given template under optimised conditions. By inference, single base alterations in the genomic template DNA may also lead to changes in the RAPD fingerprints. We have examined this potential application to detect mutations in bacteria and cultured human cells. We have utilised Escherichia coli and human lymphoblastoid cell lines exposed to UV radiation, selected for by cellular mutation assays, and compared RAPD fingerprints of mutant and non-mutant samples. Polymorphisms became evident as the presence and/or absence of DNA fragments between the two samples. A dose-dependent increase in the number of polymorphic bands was seen with UV irradiation of E. coli. To a lesser degree, polymorphisms were also evident for human lymphoblastoid DNA. The possible underlying mechanisms for these alterations in fingerprints as a result of mutation(s) in the primer binding site(s) are discussed. The ability of RAPD fingerprinting to detect a mutant in a population of non-mutants is evaluated, and whilst the lack of sensitivity inherent in the technique precludes its use as a mutation screening assay, its potential for generation of mutant and non-mutant DNA probes for other mutation detection techniques may prove to be of great merit. Teratogenesis Carcinog. Mutagen. 20:49 63, 2000. PMID- 10679750 TI - Evaluation of the genotoxic effects of pyrimethamine, an antimalarial drug, in the in vivo mouse. AB - Pyrimethamine is an antimalarial drug and a known teratogenic agent. With this drug, positive and negative results have been reported by various investigators in in vivo and in vitro genotoxicity/mutagenicity assays. In this investigation the genotoxic effects of pyrimethamine (PY) were tested in mice in vivo systems, using the bone marrow micronucleus test (MNT) and the transplacental MN test (TMNT). PY at the highest dose (40 mg/kg) induced statistically significant MN in bone marrow cells at 24 and 48 h. In the transplacental MN test, PY did not induce significant MN in fetal liver or in maternal bone marrow. Teratogenesis Carcinog. Mutagen. 20:65-71, 2000. PMID- 10679751 TI - Pathogenesis of cleft palate in mouse embryos exposed to 2,3,7, 8 tetrachlorodibenzo-p-dioxin (TCDD). AB - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces cleft palate in mouse embryos. It has been believed that TCDD inhibits palatal fusion by suppression of disappearance of medial edge epithelial (MEE) cells on palatal shelves. However, we found that exencephalic mouse embryos were resistant to the cleft palate inducing action of TCDD. In the present study, we examined cell kinetics in MEE and palatal mesenchyme in embryos exposed to TCDD with or without exencephaly for elucidation of pathogenesis of cleft palate by TCDD. Pregnant Jcl:ICR mice were given TCDD orally at 40 microg/kg at gestation day (GD) 12.5. Embryos were harvested between GD 13.5 and GD 14.5 and examined for cell kinetics by bromodeoxyuridine (BrdU) and TUNEL methods. Exencephaly was induced by intraperitoneal injection of CdCl(2) at 6 mg/kg at GD 7.5. BrdU-positive cells were decreased in TCDD-treated embryos in MEE and mesenchymal cells. TUNEL positive cells were detected in MEE both in TCDD-treated and untreated control embryos, as well as in embryos with or without exencephaly. We also measured the gap between shelves between GD 14. 0 and GD 14.5. There were no differences at GD 14.0 between control and TCDD-exposed embryos, but at GD 14.25 and GD 14.5, TCDD exposed embryos had wider gaps than controls. These findings indicate that cleft palate by TCDD results from poor development of palatal shelves. Teratogenesis Carcinog. Mutagen. 20:73-86, 2000. PMID- 10679752 TI - Assessing the predictive validity of frog embryo teratogenesis assay-Xenopus (FETAX). AB - The ability of frog embryo teratogenesis assay - Xenopus (FETAX) to identify the potential developmental toxicity of a group of diverse chemicals was evaluated by comparison with results from in vivo studies in rats. A total of 12 chemicals, three of which were shown to be teratogenic in vivo, four of which were embryolethal (but not teratogenic) in vivo, and five which did not produce any developmental toxicity in vivo in the rat were evaluated using FETAX. Results of the FETAX test with these 12 blind-coded compounds correctly predicted that three chemicals had strong teratogenic potential, four had low teratogenic hazard potential but were embryolethal, and five posed little if any developmental toxicity hazard. In addition, this study concluded that within a family of chemistry analogs could be ranked according to relative teratogenic hazard and that for the teratogenic compounds the types of malformations induced in Xenopus mimicked the abnormalities induced in vivo in rats. In summary, these results confirmed that the FETAX assay is predictive and can be useful in an integrated biological hazard assessment for the preliminary screening of chemicals. Teratogenesis Carcinog. Mutagen. 20:87-98, 2000. PMID- 10679753 TI - Analysis of recent trends in prostate cancer incidence and mortality. AB - BACKGROUND: There is debate over whether the recent increases seen in prostate cancer are due to lead-time bias from screening, or identification of clinically insignificant lesions. METHODS: Population-based incidence rates for 1973-1996 were calculated, based on the Surveillance, Epidemiology, and End Results Program (SEER) tumor registries. Relative incidence rates for prostate cancer by stage, fatal incidence, and lymph nodes were calculated, adjusted for age. RESULTS: Localized and regional stage prostate cancer increased through 1992 and then dropped. The rate of distant-stage disease was relatively stable from 1973-1991, with a decrease in distant stage starting in 1992. The 2-year mortality rates were constant for 1973-1989. A decline in the 2-year mortality among cases (fatal incidence) also began in 1992. CONCLUSIONS: These data show large increases in early disease, followed by a drop and leveling off along with a decrease in advanced disease (distant stage, 2-year mortality, positive lymph nodes). This indicates that the increasing incidence rates for prostate cancer are largely due to lead-time bias from increased early detection and treatment of prostate cancer. However, since incidence rates have not declined to rates seen in the 1970s, the additional cases may also reflect length bias from insignificant lesions or a true increase in incidence over time. PMID- 10679754 TI - Effects of tobacco smoke on tumor growth and radiation response of dunning R3327 prostate adenocarcinoma in rats. AB - BACKGROUND: The influence of tobacco smoke has been investigated on the growth rate and histology of prostate cancer, both in untreated tumors and in those subjected to fractionated irradiation. METHODS: Twenty-five rats were implanted bilaterally with Dunning R3327 tumor fragments at 10 weeks of age. Approximately 3 months later, they were randomly allocated to two groups, one of which was exposed to tobacco smoke for an hour each day, 5 days a week. Three weeks later the groups were further subdivided into two groups which acted as controls or were subjected to 5 daily doses of 6 Gy. The tumors were measured weekly to construct growth curves. At a fixed time, 9 weeks or 20 weeks later, the animals were sacrificed and the tumors were removed for histological evaluation of the tissue composition. Sections from each tumor were scored in a morphometric analysis of the fraction of the area of tumor that was occupied by (epithelial) tumor cells, by stroma, or by luminal spaces. In addition, the density of mast cells was assessed in adjacent sections stained with toluidine blue. RESULTS: Smoking caused only minor changes in the growth rates of both the control and the irradiated tumors. At the cellular level, smoking caused a small but significant increase in the fraction of tumor cells relative to controls. Irradiation also caused a small but significant decrease in tumor cell fraction compared to controls, even after 20 weeks of regrowth. This difference was reduced in the smoking and irradiation group. The main difference observed was in the mast cell numbers. Smoking caused a 4-fold increase in mast-cell density. Irradiation caused an even greater increase (25-fold). The combination of smoking and irradiation resulted in an intermediate increase (10-fold). CONCLUSIONS: Long term smoke exposure can slightly alter the growth rate and morphology of Dunning R3327 rat prostatic adenocarcinoma, but our study does not show a negative effect on the outcome of radiation treatment of this tumor model. We have also demonstrated a highly elevated number of mast cells in the irradiated group, and have shown that smoke exposure significantly depressed the radiation-induced enhancement of the number of mast cells. PMID- 10679755 TI - Heterogeneous apoptotic responses of prostate cancer cell lines identify an association between sensitivity to staurosporine-induced apoptosis, expression of Bcl-2 family members, and caspase activation. AB - BACKGROUND: The goal of this work was to identify mechanisms for the inability of metastatic prostate cancer cells to engage the apoptotic pathway following hormonal or cytotoxic therapy. METHODS: Genotypically diverse cell lines isolated from patients with metastatic disease were used. RESULTS: The LNCaP and TsuPr(1) lines exhibited quintessential apoptotic features in response to the pleiotropic apoptotic inducer staurosporine (STS): rapid cytochrome c translocation to the cytosol, proteolytic processing and catalytic activation of caspase-3 and -7, proteolytic inactivation of the death substrates DNA fragmentation factor (DFF) and poly-ADP-ribose polymerase (PARP), and TUNEL-positive polyfragmented nuclei. In contrast, DU-145 and PC-3 cells exhibited few, if any, of these features, while appearing necrotic by confocal microscopy. The presence of caspase-3 and -7 without proteolytic processing suggested that the apoptotic blockade was upstream of executioner caspases in these resistant cell lines. To identify the locus of this block, Western blot analysis of cytochrome c subcellular localization and of pro- and antiapoptotic Bcl-2 family members was performed, and suggested that heterogeneous expression of these proteins might be the underlying mechanism for apoptotic resistance to STS in these cell lines. Thus, the absence of the proapoptotic Bax in DU-145 cells indicated a mechanism for apoptotic resistance of these cells. Similarly, decreased Bax expression during STS treatment, coupled with overexpression of the antiapoptotic Bcl-x(L) and inability to translocate cytochrome c to the cytosol, provided a mechanism for the insensitivity of PC-3 cells. CONCLUSIONS: These observations suggest that activation of the apoptotic machinery in metastatic prostate cancer cell lines may be determined by expression levels of Bcl-2 family members, by the ability of cytochrome c to translocate to the cytosol, and by the ability of the caspase pathway to react in response to activation of the mitochondrial phase. PMID- 10679756 TI - Regressive changes and neuroendocrine differentiation in prostate cancer after neoadjuvant hormonal treatment. AB - BACKGROUND: We studied the extent of neuroendocrine (NE) tumor cell differentiation and its relation to regressive changes in prostate cancer after 3 month hormonal treatment. METHODS: Radical prostatectomy specimens from 103 patients, randomized to 3-month neoadjuvant LH-RH-analogue treatment (neoadjuvant group) or to surgery alone (control group), were available for analysis. The effects of hormonal treatment in terms of positive surgical margins, the degree of histopathological changes, and tumor cell proliferation were evaluated in relation to NE-differentiation assessed with antibodies against chromogranin A (CGA). RESULTS: Both the number of CGA-positive cells/cm(2) (P < 0.003) and the proportion of NE-positive tumors (P = 0.07) were greater in the neoadjuvant group than in the control group. No correlation existed between NE-differentiation and the effects of the neoadjuvant hormonal treatment; nor did NE-differentiation correlate to the decrease in serum PSA. CONCLUSIONS: Neuroendocrine differentiation in prostate cancer increases after 3 months of neoadjuvant hormonal treatment but does not correlate to the effects of hormonal treatment. PMID- 10679757 TI - Immunohistochemical staining for DNA topoisomerase II-alpha in benign, premalignant, and malignant lesions of the prostate. AB - BACKGROUND: The DNA topoisomerase II-alpha (topo II-alpha)-targeting drug etoposide was recently shown to be an active agent in the combined chemotherapy of hormone-insensitive prostatic carcinoma. Aside from being the molecular target of etoposide, topo II-alpha is also a cell proliferation marker. Much experimental data indicate that cells sensitive to topo II-targeting chemotherapeutic drugs are rapidly proliferating and show elevated topo II expression. There is little information concerning topo II expression in lesions of the prostate. METHODS: Paraffin blocks from cases of invasive prostatic carcinoma, prostatic intraepithelial neoplasia, and prostatic nodular hyperplasia were retrieved from the surgical pathology files at the University of Utah Health Sciences Center. Using a new immunohistochemical stain, specific for the alpha isoform of DNA topo II, enzyme expression was evaluated in 54 prostatic adenocarcinomas, 22 lesions of high-grade prostatic intraepithelial neoplasia (PIN), and 10 cases of benign prostatic nodular hyperplasia. Results were semiquantitated by determining for each case a topo II-alpha index, which represented the percent of positively staining cells. RESULTS: The average topo II-alpha index for well-differentiated prostatic adenocarcinomas (Gleason scores 2-4) was 1.5 +/- 0.9; for moderately differentiated tumors (Gleason scores 5-7), 3.1 +/- 2.4; and for poorly differentiated tumors (Gleason scores 8-10), 6.7 +/- 5.5. The average topo II-alpha index for all invasive prostatic adenocarcinomas was 4.0 (range, 0-19.0). Benign prostatic nodular hyperplasia had the lowest average topo II-alpha index, of 0.54 (range, 0.2-1.0). The average topo II-alpha index of 2.3 (range, 0-8.6) for high-grade prostatic intraepithelial neoplasia was intermediate between the invasive tumors and benign prostate. CONCLUSIONS: Topo II-alpha expression in carcinoma of the prostate correlates with Gleason score. The carcinomas with the highest expression of enzyme are more poorly differentiated and have the highest Gleason scores. Prostatic nodular hyperplasia shows little expression of topo II-alpha. Prostatic intraepithelial neoplasia has an average topo II-alpha index intermediate between nodular hyperplasia and carcinoma. PMID- 10679758 TI - Establishment and characterization of a prostatic small-cell carcinoma cell line (PSK-1) derived from a patient with Klinefelter syndrome. AB - BACKGROUND: Prostatic small-cell carcinoma (SMCC) is an extremely aggressive, rarely occurring tumor, and there has been no previous report of prostatic SMCC in association with Klinefelter syndrome. This study reports on the first such case and the establishment of the first cell line of SMCC from this tumor. METHODS: Prostatic SMCC tissue was derived from a 29-year-old man with Klinefelter syndrome. Characteristics of the culture tumor cells were evaluated with cell growth in vitro, neuron-specific enolase (NSE) secretion ability, tumorigenicity in nude mice, chemosensitivity to anticancer drugs, and karyotypic analysis. RESULTS: A culture cell line (PSK-1) was successfully established from prostatic SMCC with Klinefelter syndrome. PSK-1 cells had a polygonal epithelioid morphology and demonstrated loss of contact inhibition. These cells secreted NSE into the culture supernatant. Tumors produced in nude mice were histologically similar to the original SMCC. In a chemosensitivity test, PSK-1 cells were found to be sensitive in vitro to cisplatin, etoposide, and doxorubicin, but resistant to dacarbazine and 5-fluorouracil. Cytogenetic analysis showed that the PSK-1 cells at passage 35 revealed 76-84 chromosomes, with a mode of 82 chromosomes. CONCLUSIONS: PSK-1 cells could represent some properties of the original tumor cells, and could be used in studies on the etiology and treatment of this disease. PMID- 10679759 TI - Presence of receptors for bombesin/gastrin-releasing peptide and mRNA for three receptor subtypes in human prostate cancers. AB - BACKGROUND: Bombesin-like peptides can function as autocrine or paracrine growth factors and stimulate the growth of some cancer cells, including human prostate cancer. Three bombesin receptor subtypes, termed gastrin-releasing peptide receptor (GRPR), neuromedin B receptor (NMBR), and bombesin receptor subtype 3 (BRS-3), have been identified in rodents and humans. METHODS: We investigated the presence and characteristics of the functional receptors for bombesin/GRP in human prostate adenocarcinoma specimens by radio-receptor assay and the mRNA expression of the three bombesin receptor subtypes by RT-PCR. RESULTS: Of the 80 specimens of primary prostate cancer examined by receptor binding assays, 50 ( approximately 63%) showed high-affinity, low-capacity binding sites for bombesin/GRP, and 12 of these 50 receptor-positive specimens also showed a second binding site. Of the 22 prostate cancer specimens analyzed by RT-PCR, 20 (91%) expressed GRPR mRNA, 3 (14%) showed NMBR mRNA, and 2 ( approximately 9%) revealed BRS-3 mRNA. No correlation was observed between receptor expression and patients' age or pathological data. CONCLUSIONS: The detection of a wide distribution of bombesin/GRP receptors in human prostate carcinomas supports the view that they may be involved in modulation of tumor progression and suggests that approaches based on binding of bombesin receptor antagonists or new targeted cytotoxic bombesin analogs to prostate cancers could be considered for the therapy. PMID- 10679760 TI - Rye bran and soy protein delay growth and increase apoptosis of human LNCaP prostate adenocarcinoma in nude mice. AB - BACKGROUND: In this study, we investigated whether dietary intervention could inhibit tumor growth of an androgen-sensitive human prostatic cancer. METHODS: LNCaP cells were transplanted subcutaneously in nude-mice. The animals were then put on different diets and tumor take, tumor growth and prostate specific antigen (PSA) secretion were studied during 9 weeks. RESULTS: Palpable tumors developed in 75% of the tumor-cell injected sites in animals fed a control diet (corn starch, sucrose, etc.) whereas, for animals given rye bran (RB), ethyl acetate extraction from rye bran supplemented cellulose based diets (CCEE), palpable tumors were seen in only 30% and for soy protein based diets (SCC) 50% of the transplantation sites, respectively. The tumors that grew to palpable size in the rye (RB) and soy (SCC) groups were smaller and secreted less PSA than those in the control group. In the rye and soy groups tumor cell apoptosis was increased, but cell proliferation was unaffected. Addition of fat to the rye diet reduced its effect on prostate cancer growth. CONCLUSIONS: Factors in rye bran and soy protein may inhibit prostate cancer growth. The effect is more apparent for rye than for soy. Further studies are needed to identify the effective substances and to explore the mechanism of action. PMID- 10679761 TI - Follow-up ProstaScint scans verify detection of occult soft-tissue recurrence after failure of primary prostate cancer therapy. AB - BACKGROUND: A reliable imaging modality is required to uncover occult soft- tissue recurrence after failure of primary prostate cancer therapy. This retrospective study was done to evaluate the ability of the (111)Indium-labeled monoclonal antibody (ProstaScint(R)) scan in detection of prostatic bed recurrence and/or metastases to regional and/or distant lymph nodes. METHODS: One hundred sequential patients were evaluated with repeated ProstaScint(R) scans because of evidence of recurrence during the course of their disease. These 100 patients were followed closely from November 1994 and April 1999, and had concurrent bone scans and serum prostate-specific antigen (PSA) evaluations. They have had hormone therapy (n = 53) and/or experienced a rising PSA after radical prostatectomy (n = 38) or after radiation therapy (n = 56). Scan images were scored 0-3, where score 0 = negative, score 1= prostate bed uptake, score 2 = regional lymph node uptake, and score 3 = distant lymph node uptake. In each patient, the uptake of the follow-up scan(s) was compared to that of the initial scan. RESULTS: The median age was 70 years (range, 45-87), and 23 patients had a positive bone scan. The average PSA was 40.5 ng/ml (standard deviation, 223.5). There was 257 scans representing 100 patients. All patients had at least 2 scans, 35 patients had 3 scans, and 11 patients had 4 scans. No individual exhibited detectable adverse clinical reactions during or after the scan. The findings of the initial and consecutive scans were anatomically consistent in 79%, whereas in 21% there were skip metastases. In 24 patients the lesions progressed by scan and PSA, 10 patients showed progression of scan but no PSA progression, 49 patients showed no change, and 17 patients showed a remission related to adjuvant therapy. CONCLUSIONS: The consistency on repeating the scan (79%) and the high percentage of patients showing persistent uptake at the prostate bed (43%) as well as the percentage of detection of regional nodes (20%) and distant nodes (32%) reflects the importance of using the ProstaScint(R) scan in finding occult recurrences after primary treatment failure of prostate cancer. These results are similar to those reported earlier in autopsy series studies in similar populations. PMID- 10679763 TI - Preface PMID- 10679762 TI - Serum levels of PSMA. PMID- 10679764 TI - Stem cells and the cellular organization of the brain. AB - The modern field of neuroscience emerged in the hands of neuroanatomists at the end of the last century. They recognized that information flows through arrays of cells and that the structure of the cells shapes the passage of information through the brain. In the middle of this century, the role of ion flux in the movement of information along axons was established. The electron microscope and the microelectrode helped to explain information transfer at the synapse, and study of the modulation of synaptic strength is currently a major area of neuroscience research. The origin of the many types of neuron seen by the early anatomists remains enigmatic, but the solution to this mystery is now emerging. The identification of stem cells and of the mechanisms that control their differentiation into distinct neuron types will contribute to a new understanding of both the origin of neuronal types and neuronal circuits. The current enthusiasm for stem cells is stimulated by interest from both the academic and nonacademic communities. These enthusiasts recognize a simple truth, that the cells are smart; they are the agents that control cell number and type in the brain. They create the structure that generates the higher order brain function. If these premises are true, then in the new century our understanding of the nervous system will be transformed by the facts of stem cell biology. J. Neurosci. Res. 59:298-300, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10679765 TI - Early cortical precursors do not undergo LIF-mediated astrocytic differentiation. AB - Multipotential stem cells have been isolated from the developing and adult CNS. Similar identified factors control the differentiation of these cells. A striking example is the instructive action of CNTF/LIF activating the JAK/STAT pathway to induce astrocytic differentiation in both fetal and adult CNS stem cells. Here we show that E12 cortical precursors express functional LIF receptors but do not exhibit this differentiation response to CNTF/LIF either in explant or in dissociated cell culture. The lack of response to LIF-induced astrocytic differentiation is maintained in cocultures with LIF responsive cells derived from E15 cortex. This suggests cell intrinsic differences between early and late stage precursors in the interpretation of LIF-mediated signaling; however, the early nestin-positive precursor population differentiates into both neurons and neural crest derivatives. These data define differences between CNS stem cells from different stages of cortical development. J. Neurosci. Res. 59:301-311, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10679766 TI - Developmental changes in neural progenitor cell lineage commitment do not depend on epidermal growth factor receptor signaling. AB - Multipotent neural progenitor cells become progressively more biased towards a glial fate during development coincident with an increase in expression of the epidermal growth factor receptor (EGFR). To determine whether differences in lineage commitment of neural progenitor cells from different stages are causally related to expression of the EGFR and whether generation of glia is EGFR dependent, we used an EGFR-specific tyrosine kinase inhibitor, PD158780, to block the activation of EGFR in progenitor cells. Treatment of cultured neonatal progenitor cells with PD158780 completely blocked EGF-induced proliferation of the cells but did not affect bFGF-induced proliferation. Nevertheless, treatment with the inhibitor failed to inhibit the generation of astroglia in the presence of either EGF or bFGF. Treatment with bone morphogenetic protein-2 (BMP2) enhanced astroglial differentiation and suppressed oligodendroglial (OL) differentiation. PD158780 treatment had no effect on the BMP2-induced astroglial differentiation or OL suppression. These observations suggest that the generation of astroglia is not dependent on EGFR activation. Because it was still possible that the progenitor cell responses reflected a prior history of EGFR signaling, rat forebrain cells were cultured in the presence of PD158780 from a time (E12.5) preceding expression of the EGFR. After time in culture, the E12.5 cells expressed EGFR by Western analysis both in the presence and in the absence of PD158780, but activation of EGFR kinase (receptor autophosphorylation) was undetectable in the presence of PD158780 and the cells did not proliferate in response to EGF. Nevertheless, astroglial differentiation was normal in PD158780 treated cells both in the absence and in the presence of BMPs or CNTF. Furthermore, the propensity towards glial differentiation increased with time in culture even in the absence of EGFR signaling. This suggests that the increased bias towards glial differentiation during development does not depend on EGFR signaling. PMID- 10679768 TI - Endogenous IGF-1 regulates the neuronal differentiation of adult stem cells. AB - Stem cells from the adult forebrain of mice were stimulated to form clones in vitro using fibroblast growth factor-2 (FGF-2). At concentrations above 10 ng/ml of FGF-2, very few clones gave rise to neurons; however, if FGF-2 was removed after 5 days, 20-30% of clones subsequently gave rise to neurons. The number of neuron-containing clones and the number of neurons per clone was significantly enhanced, if insulin-like growth factor (IGF)-1 or heparin were added subsequent to FGF-2 removal. The spontaneous production of neurons after FGF-2 removal was shown to be due to endogenous IGF-1, since antibodies to IGF-1 and an IGF-1 binding protein totally inhibited neuronal production. Similarly, these reagents also abrogated the neuron-promoting effects of heparin. Thus, it appears that endogenous IGF-1 may be a major regulator of stem cell differentiation into neurons. Furthermore, it was found that high levels of IGF-1 or insulin promoted the maturation and affected the neurotransmitter phenotype of the neurons generated. PMID- 10679767 TI - Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone. AB - Adult humans, like their nonhuman mammalian counterparts, harbor persistent neural progenitor cells in the forebrain ventricular lining. In the absence of adequate surface markers, however, these cells have proven difficult to isolate for study. We have previously identified and selected neural progenitor cells from both the fetal and adult rodent ventricular zone (VZ), by sorting forebrain cells transfected with plasmid DNA encoding the gene for green fluorescent protein driven by the early neuronal promoter for Talpha1 tubulin (P/Talpha1:hGFP). We have now extended this approach by purifying both P/Talpha1:hGFP tubulin-defined neuronal progenitors, as well as potentially less committed E/nestin:hGFP-defined neural progenitor cells, from the adult human VZ. The ventricular wall of the temporal horn of the lateral ventricle was dissected from temporal lobes obtained from four adult patients undergoing therapeutic lobectomy. These samples were dissociated, and the cultured cells transduced with either P/Talpha1:hGFP or E/nestin:EGFP plasmid DNA. A week later, the cells were redissociated, selected via fluorescence-activated cell sorting (FACS) on the basis of neural promoter-driven GFP expression, and replated. The majority of these cells expressed the early neuronal protein betaIII-tubulin upon FACS; within the week thereafter, most matured as morphologically evident neurons that coexpressed betaIII-tubulin and microtubule-associated protein (MAP)-2. Many of these neurons had incorporated bromodeoxyuridine in vitro in the days before FACS, indicating their mitogenesis in vitro. Thus, the use of fluorescent transgenes under the control of early neural promoters permits the enrichment of neuronal progenitor cells from the adult human ventricular zone. The specific acquisition, in both purity and number, of residual neural progenitor cells from the adult human brain may now permit hitherto unfeasible studies of both their biology and practical application. PMID- 10679769 TI - Motoneuron differentiation of immortalized human spinal cord cell lines. AB - Human motoneuron cell lines will be valuable tools for spinal cord research and drug discovery. To create such cell lines, we immortalized NCAM(+)/neurofilament(+) precursors from human embryonic spinal cord with a tetracycline repressible v-myc oncogene. Clonal NCAM(+)/neurofilament(+) cell lines differentiated exclusively into neurons within 1 week. These neurons displayed extensive processes, exhibited immunoreactivity for mature neuron specific markers such as tau and synaptophysin, and fired action potentials upon current injection. Moreover, a clonal precursor cell line gave rise to multiple types of spinal cord neurons, including ChAT(+)/Lhx3(+)/Lhx4(+) motoneurons and GABA(+) interneurons. These neuronal restricted precursor cell lines will expedite the elucidation of molecular mechanisms that regulate the differentiation, maturation and survival of specific subsets of spinal cord neurons, and the identification and validation of novel drug targets for motoneuron diseases and spinal cord injury. PMID- 10679770 TI - Trophic factors: An evolutionary cul-de-sac or door into higher neuronal function? AB - Trophic factors, such as the neurotrophins, CNTF, and GDNF, represent unique families of proteins that are essential for the development of the vertebrate nervous system. Surprisingly, there is little evidence to date that these proteins exist in Drosophila melanogaster and Caenorhabditis elegans, even though other polypeptide growth factors, such as EGF, FGF, and insulin, are conserved in these species. For the neurotrophins, the evolution of NGF, BDNF, NT-3, and NT-4 as a family implies that these signaling molecules may be involved in mediating additional neural activities, such as learning, memory, and behavior. Indeed, there is abundant evidence now that BDNF is involved in synapse modification, neurotransmitter release, long-term potentiation, and mechanosensation. The widening scope of neurotrophin action will require more physiological, genomic, and integrative approaches to understand fully the mechanisms by which survival factors can mediate so many diverse effects. PMID- 10679771 TI - Association of the Abl tyrosine kinase with the Trk nerve growth factor receptor. AB - Nerve growth factor (NGF) initiates the majority of its biological effects by promoting the dimerization and activation of the tyrosine kinase receptor TrkA. In addition to rapid increases in the phosphorylation of phosphatidylinositol 3' kinase (PI 3-kinase) and phospholipase C-gamma and increased ras activity, phosphorylation of c-Crk and paxillin proteins has been observed upon TrkA activation. The c-Abl tyrosine kinase is involved in the control of the axonal cytoskeleton and is known to interact with c-Crk proteins. Here we have tested the possibility that TrkA receptors might form an association with the c-Abl protein. After transfection in 293T cells, TrkA and c-Abl kinases could be coimmunoprecipitated. This interaction did not require TrkA receptors to be autophosphorylated. Mapping analysis indicated that the region of c-Abl association was confined to the juxtamembrane region of TrkA. The interaction of c-Abl with TrkA was also observed in differentiated pheochromocytoma PC12 cells. These results suggest that c-Abl may be recruited to the NGF receptor complex and be involved in regulating specific phosphorylation events that occur during neuronal differentiation. PMID- 10679772 TI - Differential effects of TrkC isoforms on sensory axon outgrowth. AB - Neurotrophins are powerful regulators of neuronal morphology. Several lines of evidence are consistent with the idea that characteristic axonal and dendritic morphologies throughout the nervous system may be determined by local patterns of neurotrophin and neurotrophin receptor expression. Neurotrophin receptor tryosine kinases (Trks) exist in both tyrosine-containing (TK+) and tyrosine-lacking (TK-) isoforms, both of which are expressed in many neuronal populations. However, ratios of TK+ to TK- isoforms may vary at different stages of development and may be differentially distributed to cellular compartments. To test whether these isoforms have different functions related to axon outgrowth, full-length or tyrosine kinase-lacking TrkC receptors were overexpressed in embryonic dorsal root ganglion neurons maintained in explant cultures in neurotrophin-3 (NT-3) containing media. Neurons were transfected with plasmid DNA encoding enhanced yellow fluorescent protein (EYFP) and TrkC receptor isoforms by particle-mediated gene transfer. Control neurons possessed 3.7 +/- 1.3 primary processes and 113.8 +/- 46 branch points. About 80% of the branches were located along the distal part of the axon. Transfection with the trkC TK+ increased the number of primary processes (6.5 +/- 2.8), whereas transfection with trkC TK- reduced the formation of primary processes (3.0 +/- 1.3). Surprisingly, the distribution of branch points was shifted to the proximal region of axons in neurons transfected with trkC TK-. These observations are consistent with the idea that differential expression of Trk isoforms during development may sculpt axonal morphology. PMID- 10679773 TI - TrkB expression and early sensory neuron survival are independent of endogenous BDNF. AB - Sensory neurons initially survive independently of neurotrophins in culture during the stage of development when their axons are growing to their targets. Because mRNAs encoding brain-derived neurotrophic factor (BDNF) and its receptor tyrosine kinase TrkB are detectable in subsets of sensory neurons from the earliest stages of their development, we investigated whether a BDNF autocrine loop is responsible for sustaining the survival of these neurons during this early stage in their development. Low-density dissociated cultures of nodose and dorsal root ganglion neurons were established from wild type and BDNF(-/-) mouse embryos at this stage and were grown in defined medium without added neurotrophins. Wild type and BDNF-deficient neurons survived equally well under these conditions, indicating that a BDNF autocrine loop does not play a role in sustaining the survival of sensory neurons during the earliest stages of their development. As sensory axons approach their targets, TrkB expression increases in a subset of neurons that becomes dependent on BDNF produced by other cells. Because numerous studies have shown that neurotrophins, including BDNF, increase expression of their receptors, we investigated whether endogenous BDNF is required for the increase in TrkB expression observed during stage of development. Quantitative reverse transcriptase-polymerase chain reaction (RT PCR) showed that the developmental increase in TrkB mRNA expression occurred normally in the sensory ganglia of BDNF(-/-) embryos. Taken together, our studies of sensory neuron development in BDNF-deficient embryos have demonstrated that endogenous BDNF is neither required for the early survival of these neurons nor for the induction of TrkB expression. PMID- 10679774 TI - Reduction of endogenous TGF-beta increases proliferation of developing adrenal chromaffin cells in vivo. AB - Chromaffin cells and sympathetic neurons are derivatives of the sympathoadrenal cell lineage of the neural crest. Although they are similar with respect to many cell biological aspects, chromaffin cells, in contrast to sympathetic neurons, continue to synthesize DNA and proliferate through their whole life span. Large numbers of neural and hormonal signals have been implicated in the regulation of chromaffin cell proliferation based on in vitro studies. We have previously shown that chromaffin cells synthesize and release transforming growth factor-beta (TGF beta) and that exogenously applied TGF-beta suppresses chromaffin cell proliferation in vitro. We show now that TGF-beta is also a physiologically relevant factor in the control of cell division in developing chromaffin cells. We have neutralized endogenous TGF-beta in quail embryos using a monoclonal antibody recognizing all three TGF-beta isoforms, TGF-beta1, -beta2, and -beta3. Embryos deprived of TGF-beta show a prominent increase in numbers of tyrosine hydroxylase (TH)-immunoreactive adrenal chromaffin cells and TH-positive cells incorporating 5'-bromo-2'deoxyuridine. This is the first documentation of the physiological significance of a factor that has been suggested to play a role in the regulation of chromaffin cell mitosis based on in vitro experiments. PMID- 10679775 TI - Development of the interstitial cell of Cajal: origin, kit dependence and neuronal and nonneuronal sources of kit ligand. AB - Kit is a marker for interstitial cells of Cajal (ICC). ICCs interact with enteric neurons and are essential for gastrointestinal motility. The roles of neural crest-derived cells, neurons, Kit, and Kit ligand (KL) in ICC development were analyzed. ICC development lagged behind that of neurons and smooth muscle. Although mRNA encoding Kit and KL was detected at E11, Kit-immunoreactive ICCs did not appear until E12 in foregut and E14 in terminal hindgut. Transcripts of Kit and KL and Kit-immunoreactive cells were found in aganglionic gut from ls/ls and c-ret -/- mice. ICCs also developed in crest-free cultures of ls/ls terminal colon. ICCs appeared in cultures of noncrest- but not those of crest-derived cells isolated from the fetal bowel by immunoselection with antibodies to p75(NTR). KL immunoreactivity was coincident in cells with neuronal or smooth muscle markers. The development of ICCs in cultures of mixed cells dissociated from the fetal gut was dependent on plating density. No ICCs appeared at /=200,000 cells/ml. Exogenous KL partially substituted for a high plating density. These data support the ideas that mammalian ICCs are neither derived from the neural crest nor developmentally dependent on neurons. ICC differentiation/survival requires KL, which can be provided by neurons or cells in a smooth muscle lineage. Neurons may be needed for development of myenteric ICCs and the mature ICC network. PMID- 10679776 TI - Neurite outgrowth can be modulated in vitro using a tetracycline-repressible gene therapy vector expressing human nerve growth factor. AB - The delivery of neurotrophic factors to the adult nervous system has potential applications for the treatment of neurodegenerative diseases and trauma. In vivo and ex vivo gene therapy offer a means of delivering growth factors and other therapeutic substances to the central nervous system (CNS) in an intraparenchymal, accurately targeted, and regionally restricted manner. Ideally, gene therapy delivery systems should also be regulatable, allowing exogenous control of amount of gene product delivery. In the present experiment, a tetracycline-regulatable gene expression system was generated to determine whether controllable release of nerve growth factor (NGF) and green fluorescent protein (GFP) from primary rat fibroblasts could modulate biological responses (neurite outgrowth) in vitro. Using a tetracycline-repressible construct, it was found that NGF mRNA, NGF protein, and NGF-induced neurite outgrowth could be tightly regulated within a 24 hour period, and in a dose-dependent fashion, by exposure to the tetracycline analog doxycycline. Similarly, levels of green fluorescence could be regulated in GFP-transfected cells. These findings in a neurobiological system lay the framework for future studies using regulated neurotrophin delivery in in vivo models of neurodegenerative diseases and CNS injury. PMID- 10679777 TI - Glial differentiation and lineages. PMID- 10679778 TI - Local control of oligodendrocyte development in isolated dorsal mouse spinal cord. AB - The earliest oligodendrocyte precursors have been proposed to arise in the ventral ventricular zone of the embryonic thoraco-lumbar spinal cord and subsequently migrate to populate dorsal spinal cord. Using the expression of O4 immunoreactivity to define cells of the oligodendrocyte lineage, the development of oligodendrocytes in different regions of the mouse spinal cord was assayed. Consistent with earlier studies in other species, isolated explants of E11 ventral but not dorsal mouse spinal cord developed oligodendrocytes after 7 days in vitro. In contrast, in cultures derived from E13 embryos O4(+) oligodendrocytes developed in both ventral and dorsal cultures after 5 days in vitro. These data are consistent with a ventral to dorsal migration of committed oligodendrocyte progenitors occurring between E11 and E13. Although isolated early embryonic dorsal spinal cord does not give rise to oligodendrocytes in short term cultures, in long term cultures O4(+) cells develop in a subset of dorsal explants. After 10 days in vitro approximately 25% of both cervical and thoraco-lumbar E11 derived dorsal explants contained significant numbers of O4(+) cells. The molecular requirements for the dorsally-derived oligodendrocytes was similar to that in ventral cord. The appearance of O4(+) cells was dependent on sonic hedgehog and enhanced by neuregulin. These data suggest that early embryonic dorsal mouse spinal cord has an independent potential to generate oligodendrocytes under appropriate conditions. Whether this potential is realized during normal spinal cord development is currently unknown. PMID- 10679779 TI - Tracing human oligodendroglial development in vitro. AB - Human neural precursor cell cultures (neurospheres) were established from fetal brain tissues of 15-20 gestation weeks and propagated for over a year in the presence of epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor. Neurospheres were differentiated without the presence of above growth factors to follow the development of oligodendroglia. Oligodendroglial progenitors, identified by their bipolar morphology and expression of platelet derived growth factor receptor-alpha (PDGFRalpha), emerged from spheres as early as 1 DIV; O4+ cells with bipolar to multipolar processes were observed at 3 DIV whereas O1+ multiprocess-bearing oligodendroglia did not appear until 5-7 DIV. They further differentiated to myelin basic protein-expressing oligodendrocytes after 2-3 weeks in culture. Thus, human oligodendroglial maturation in vitro follows the same pathway as rat cells but takes twice as long as their rodent counterparts. Bromodeoxyuridine incorporation indicated that PDGFRalpha expressing cells but not O4+ oligodendroglia proliferated. More oligodendroglial progenitors incorporated BrdU and more O4+ cells survived when they were in contact with neurons and astrocytes than when they developed beyond the astrocyte layer. In addition, oligodendroglia on astrocytes had a complex process branching whereas those growing beyond astrocyte layer often formed membrane sheaths. Thus the survival, proliferation and maturation of oligodendroglia are influenced by other cell types. PMID- 10679780 TI - Analysis of oligodendroglial differentiation using cDNA arrays. AB - We used cDNA arrays to investigate molecular aspects of the differentiation of an immortalized line of oligodendroglial progenitors, and of immunopan-purified primary cultures of oligodendroglial progenitors, to immature oligodendroglia. Developmental regulation of the proteolipid and 2-hydroxyacylsphingosine 1 galactosyltransferase genes was tighter in the primary than in the immortalized cells. Our data suggest that increased expression of genes encoding the following proteins are involved in oligodendroglial differentiation: Fyn, Erk, p85, G-alpha 12 guanine nucleotide binding, and transducin beta-2 signal transduction molecules; glial maturation factor; the proteasomal subunits C8 and C3; the proteasomal targeting molecule polyubiquitin; the cell cycle regulatory proteins Set, protein phosphatase 2A, and nuclear tyrosine phosphatase (PRL-1); and the high-affinity glutamate cotransporter EAAC-1. PMID- 10679781 TI - Oligodendrocytes as glucocorticoids target cells: functional analysis of the glycerol phosphate dehydrogenase gene. AB - Previous research has established that the development and function of oligodendrocytes are influenced by glucocorticoids. The enzyme glycerol phosphate dehydrogenase (E.C.1.1.1.8) has been used as a model to study glucocorticoid regulation of gene expression in oligodendrocytes and the C6 glial cell line. In the rat brain this enzyme is exclusively localized to oligodendrocytes. The sequence of the 5' flanking region for the rat gene encoding Glycerol Phosphate Dehydrogenase (GPDH; EC 1.1.1.8) was determined. 4 kb of sequence from the 5' flanking region, exon 1, and part of intron 1 of the rat GPDH gene was compared to the corresponding mouse sequence. Dotplot matrix comparison revealed that the rat sequence is more than 80% similar to the mouse sequence, but differs from the mouse sequence in two regions: the rat sequence is devoid of 200 bp of B1 repeat sequence that is present in the mouse, and the rat sequence has an excess 700 bp of B2 repeat sequence inserted between -0.7 kb and -1. 4 kb that is absent in the mouse. To determine the regulatory activity of the rat GPDH 5' flanking region, various portions of the rat GPDH 5' flanking region were placed in luciferase reporter constructs and tested for transcriptional activity. Transient transfection of reporter constructs into the C6 glial cell line revealed that the distal end of the 5' flanking region was glucocorticoid-inducible. A 385 bp Glucocorticoid Response Unit (GRU) was identified whose glucocorticoid induction was enhanced by dibutyryl-cAMP and reduced by phorbol esters. Sequence analysis of the GRU revealed the presence of four consensus GRE sequences and other putative consensus elements. Results here suggest that the 5' flanking region of the GPDH gene mediates the ligand-inducible regulation of GPDH, and that multiple signaling pathways converge at the 5' regulatory sequence to modulate GPDH gene expression in oligodendrocytes. PMID- 10679782 TI - Expression of rab GTP-binding proteins during oligodendrocyte differentiation in culture. AB - Oligodendrocytes (OLs) synthesize and transport vast amounts of proteins and lipids from the cell body to the morphologically and biochemically distinct domains of the myelin membrane. From our prediction that regulators of vesicular transport should be up-regulated at the time of myelin production, we hypothesized that the up-regulated and unidentified small GTPases found by Huber et al. [1994a] may be Rab proteins. We have analyzed the mRNA expression of rabs in OLs, and have detected rabs 10, 11b, 18, 24, 26, and 28 in addition to rabs that were found previously. Our data show that among the Rabs so far detected during differentiation, only Rabs 5a and 8a exhibited up-regulation in addition to the previously published Rab3a (Madison et al. [1999], J. Neurochem. 72:988 998). We discuss the limited extent of up-regulation of rabs in the context of the presumed necessity for an increase in Rab activity during myelin assembly. PMID- 10679783 TI - Localization of brain-derived neurotrophic factor and TrkB receptors to postsynaptic densities of adult rat cerebral cortex. AB - Although neurotrophins are critical for neuronal survival and differentiation, recent studies suggest that they also regulate synaptic plasticity. Brain-derived neurotrophic factor (BDNF) rapidly increases synaptic transmission in hippocampal neurons, and enhances long-term potentiation (LTP), a cellular and molecular model of learning and memory. Loci and precise mechanisms of BDNF action remain to be defined: evidence supports both pre- and postsynaptic sites of action. To help elucidate the synaptic mechanisms of BDNF action, we used antisera directed against the extracellular and intracellular domains of trkB receptors, anti trkBout and anti-trkBin, respectively, to localize the receptors in relation to synapses. Synaptic localization of BDNF was examined in parallel using anti-BDNF antisera. By light microscopy, trkBin and trkBout immunoreactivities were localized to hippocampal neurons and all layers of the overlying visual cortex. Immunoelectron microscopic analysis of the cerebral cortex revealed that trkB and BDNF localize discretely to postsynaptic densities (PSD) of axo-spinous asymmetric synaptic junctions, that are the morphological correlates of excitatory, glutamatergic synapses. TrkB immunoreactivity was also detected in the nucleoplasm by light and electron microscopy. Western blot analysis indicated that both anti-trkBout and anti-trkBin antisera react with a protein band in the PSD corresponding to the molecular weight expected for trkB; however, molecular species distinct from that for trkB were recognized in the nuclear fraction by both anti-trkBin and anti-trkBout antisera, indicating that the nuclear immunoreactivity, seen by immunocytochemistry, reflects cross-reactivity with proteins closely related to, but distinct from, trkB. The PSD localization of both BDNF and trkB supports the contention that this receptor/ligand pair participates in postsynaptic plasticity. PMID- 10679784 TI - Obituary PMID- 10679785 TI - Spatiotemporal development of oligodendrocytes in the embryonic brain. AB - In the central nervous system (CNS), oligodendrocytes have long been considered to be the last cell type to be generated during development. In rodents, the progenitor cells that give rise to oligodendrocytes have been reported to originate in the subventricular zone. Here, we review recent data demonstrating the existence of oligodendrocyte precursor cells in the ventricular layer of the neural tube that emerge prior to the progenitor stage. Oligodendrocyte precursors arise in restricted foci that are distributed along the rostrocaudal axis of the neural tube, for the most part ventrally. The generation of oligodendrocyte precursor cells occurs either simultaneously with, or follows closely upon the emergence of the first neurons. Experiments with quail-chick chimeras provide evidence that oligodendrocyte progenitors derived from ventricular precursors migrate either tangentially or radially to colonize extensive or segmentally restricted territories of the brain. The choice depends on their site of origin. Finally, we discuss the possibility that oligodendrocytes could be a mosaic population that originates from at least two types of precursor cells. PMID- 10679786 TI - Oligodendrocytes and the control of myelination in vivo: new insights from the rat anterior medullary velum. AB - The rat anterior medullary velum (AMV) is representative of the brain and spinal cord, overall, and provides an almost two-dimensional preparation for investigating axon-glial interactions in vivo. Here, we review some of our findings on axon-oligodendrocyte unit relations in our adult, development, and injury paradigms: (1) adult oligodendrocytes are phenotypically heterogeneous, conforming to Del Rio Hortega's types I-IV, whereby differences in oligodendrocyte morphology, metabolism, myelin sheath radial and longitudinal dimensions, and biochemistry correlate with the diameters of axons in the unit; (2) oligodendrocytes derive from a common premyelinating oligodendrocyte phenotype, and divergence of types I-IV is related to the age they emerge and the presumptive diameter of axons in the unit; (3) during myelination, axon oligodendrocyte units progress through a sequence of maturation phases, related to axon contact, ensheathment, establishment of internodal myelin sheaths, and finally the radial growth and compaction of the myelin sheath; (4) we provide direct in vivo evidence that platelet-derived growth factor-AA (PDGF-AA), fibroblast growth factor (FGF-2), and insulin-like growth factor-I (IGF-I) differentially regulate these events, by injecting the growth factors into the cerebrospinal fluid of neonatal rat pups; (5) in lesioned adult AMV, transected central nervous system (CNS) axons regenerate through the putatively inhibitory environment of the glial scar, but remyelination by oligodendrocytes is incomplete, indicating that axon-oligodendrocyte interactions are defective; and (6) in the adult AMV, cells expressing the NG2 chondroitin sulphate have a presumptive adult oligodendrocyte progenitor antigenic phenotype, but are highly complex cells and send processes to contact axolemma at nodes of Ranvier, suggesting they subserve a specific perinodal function. Thus, axons and oligodendrocyte lineage cells form interdependent functional units, but oligodendrocyte numbers, differentiation, phenotype divergence, and myelinogenesis are governed by axons in the units, mediated by growth factors and contact-dependent signals. PMID- 10679787 TI - Hepatocyte growth factor protects cultured rat cerebellar granule neurons from apoptosis via the phosphatidylinositol-3 kinase/Akt pathway. AB - Recent studies suggest that hepatocyte growth factor (HGF) functions as a neurotrophic factor in the central nervous system. In this study, we investigated the neuroprotective effect of HGF and its mechanism of action. We used cultured cerebellar granule neurons that underwent apoptosis when the culture medium was changed from that containing serum with 25 mM K(+) to serum-free medium containing 5 mM K(+), and HGF prevented apoptotic cell death. HGF stimulated both mitogen-activated protein (MAP) kinase and phosphatidylinositol-3 (PI3)-kinase activity in cerebellar granule neurons. Two specific inhibitors of PI3-kinase, wortmannin and LY294002, efficiently blocked this neuroprotective effect of HGF. In contrast, PD98059, a selective inhibitor of MAP kinase kinase (MEK), did not affect the anti-apoptotic effect of HGF. The downstream signal of PI3-kinase in this protection was further investigated. HGF-induced phosphorylation of Akt and pretreatment of the cells with wortmannin completely impaired Akt activation. These results suggest that HGF prevents apoptosis in cerebellar granule neurons via the PI3-kinase/Akt pathway. PMID- 10679788 TI - Neuroprotective effects of a new glutathione peroxidase mimetic on neurons of the chick embryo's retina. AB - During their period of naturally occurring neuronal death, retinal ganglion cells are particularly vulnerable to axotomy. The resulting cell death requires protein synthesis and is redox-regulated, since antioxidants protect axotomized-ganglion cells when given in doses that maintain the redox status near an optimal set point. Here we report the effects of BXT-51072, a new glutathione peroxidase mimetic, on ganglion cell death induced in various ways in the retinas of chick embryos. The intraocular injection of BXT-51072 protected axotomized neurons at doses in a narrow (tenfold) range. It also reduced the deleterious effects of intraocular tert-butyl hydroperoxide, an inducer of lipid peroxidation, and diminished the excitotoxic degeneration induced by N-methyl-D-aspartate. However, BXT-51072 did not noticeably reduce naturally occurring cell death. Globally, our results show that BXT-51072 has numerous protective effects in the retina. In accordance with published data, the present report indicates that glutathione peroxidase mimetics may have potential applications for neurologic or degenerative diseases. PMID- 10679789 TI - Differentiation of rat striatal embryonic stem cells in vitro: monolayer culture vs. three-dimensional coculture with differentiated brain cells. AB - Several groups have demonstrated the existence of self-renewing stem cells in embryonic and adult mouse brain. In vitro, these cells proliferate in response to epidermal growth factor, forming clusters of nestin-positive cells that may be dissociated and subcultured repetitively. Here we show that, in stem cell clusters derived from rat embryonic striatum, cell proliferation decreased with increasing number of passages and in response to elevated concentrations of potassium (30 mM KCl). In monolayer culture, the appearance of microtubule associated protein type-5-immunoreactive (MAP-5(+)) cells (presumptive neurons) in response to basic fibroblast growth factor (bFGF) was reduced at low cell density and with increasing number of passages. In the presence of bFGF, elevated potassium caused a more differentiated neuronal phenotype, characterized by an increased proportion of MAP-5(+) cells, extensive neuritic branching, and higher specific activity of glutamic acid decarboxylase. Dissociated stem cells were able to invade cultured brain cell aggregates containing different proportions of neurons and glial cells, whereas they required the presence of a considerable proportion of glial cells in the host cultures to become neurofilament H positive. The latter observation supports the view that astrocyte-derived factors influence early differentiation of the neuronal cell lineage. PMID- 10679790 TI - Multilamellar packing of myelin modeled by lipid-bound MBP. AB - Membrane compaction and adhesion at the major dense line (cytoplasmic apposition) of myelin, particularly in the central nervous system (CNS), is typically attributed to myelin basic protein (MBP). To explore the role of MBP in myelin membrane adhesion, we attempted to reconstitute the major dense line of myelin from purified lipid-bound MBP, which is a detergent-soluble form of MBP that retains the binding of all the myelin lipids. Removal of detergent by long-term dialysis yielded a precipitate, which, when analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and thin-layer chromatography, contained MBP that was still associated with myelin lipids, but in different proportions than in the native membrane. Comparison of lipid composition among isolated myelin, MBP-free myelin lipids, and lipid-bound MBP aggregates showed that the lipid-bound form of the protein was specifically enriched in phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, phosphatidylinositol, and phosphatidylserine. Electron microscopy and x-ray diffraction demonstrated that the lipid-MBP complexes formed multilayers having periods of 70-85 A, which correspond in width to individual myelin membranes. By contrast, the lipids alone assembled as multilayers having a period of approximately 40 A. Thus, the detergent-soluble form of MBP, which is bound to lipids, might serve as a simple model for the cytoplasmic apposition of myelin. PMID- 10679791 TI - Visualization of beta-amyloid peptide (Abeta) phagocytosis by human mononuclear phagocytes: dependency on Abeta aggregate size. AB - Previous studies from this laboratory have shown that human monocytes exposed to beta-amyloid peptide (Abeta) exert a graded neurotoxic response in an organotypic brain culture paradigm. Moreover, this toxicity can be reduced by compounds that inhibit cell motility and phagocytosis, suggesting that internalization of Abeta may be a requirement for neurotoxic action. To confirm that Abeta is indeed phagocytosed by monocytes and to further lay the groundwork for resolving the possible linkage between this process and neurotoxicity, we examined Abeta:monocyte interactions using immunocytochemistry and fluorescence histochemistry followed by confocal microscopy and three-dimensional image reconstruction. Internalization of Abeta was detected by 24 hr following exposure of monocytes to the purified peptide, and the relative efficacy of this process appeared to be influenced by the size of the Abeta aggregates. Specifically, smaller aggregates were observed to be more efficiently internalized, while larger Abeta masses tended to reside only on the monocyte surface, apparently bound to several monocytes at once. Both colchicine and cytochalasin D, cytoskeletal perturbants that block phagocytosis, caused Abeta to accumulate in deep pits within monocytes and inhibited complete envelopment by monocyte cytoplasm. These results suggest that monocytes can indeed phagocytose aggregates of Abeta and that this process may be critical in activating these cells to a neurocytopathic state. Accordingly, interference of Abeta phagocytosis by monocytes or monocyte-derived cells may be a novel target for therapeutic action. PMID- 10679792 TI - A panel of epitope-specific antibodies detects protein domains distributed throughout human alpha-synuclein in Lewy bodies of Parkinson's disease. AB - To facilitate studies of the normal biology of alpha-synuclein, a member of a family of neuronal proteins of unknown function, and to elucidate the role of alpha-synuclein pathologies in neurodegenerative diseases, we generated and characterized a panel of anti-synuclein antibodies. Here we demonstrate that these antibodies recognize defined epitopes spanning the entire length of human alpha-synuclein, and that some of these antibodies also cross-react with zebra finch and rodent synucleins. Since alpha-synuclein has been reported to be a major component of Lewy bodies (LBs) in Parkinson's disease (PD), dementia with LBs and common variants of Alzheimer's disease, we performed immunohistochemical studies showing that these antibodies label numerous LBs in the PD substantia nigra, thereby localizing protein domains throughout human alpha-synuclein in LBs. Taken together, our data indicate that this panel of antibodies can be exploited to probe the normal biology of alpha-synuclein as well as the role of pathological forms of this protein in PD and related neurodegenerative synucleinopathies. PMID- 10679793 TI - Lobster GABA receptor subunit expressed in neural tissues. AB - A cDNA encoding an ionotropic gamma-aminobutyric acid (GABA) receptor subunit was isolated from a lobster (Homarus americanus) cDNA library. A longer version of this cDNA, containing a 108-bp insert, was also detected. The two cDNAs are predicted to encode alternatively spliced proteins of 485 and 521 amino acids, respectively. The sequences were most similar to the Drosophila RDL (resistance to dieldrin) GABA subunit with 54% identity, and 30-35% identity with vertebrate ionotropic GABA receptor subunits. Only the shorter clone formed functional ion channels when transfected into human embryonic kidney (HEK) 293 cells. GABA caused a Cl(-)-selective current in the presence of GABA that was blocked by picrotoxin. The GABA-induced current was weakly sensitive to the GABA(A) antagonist, bicuculline, but was enhanced by pentobarbital. Expression of the GABA receptor mRNA was highest in brain and the olfactory organ, but was not detected in leg muscle. These data suggest that the isolated cDNAs are likely to encode proteins that comprise subunits of native GABA receptors expressed in olfactory receptor neurons and projection neurons of the olfactory deutocerebrum. PMID- 10679794 TI - Influence of transferrin on manganese uptake in rat brain. AB - To evaluate the influence of transferrin (Tf) on manganese (Mn) uptake in the brain, pH 8.6 buffer-treated (54)MnCl(2), which has a higher affinity for Tf than untreated (54)MnCl(2), and Tf-bound (54)Mn were prepared. When pH 8.6 buffer treated (54)MnCl(2) and untreated (54)MnCl(2) were incubated with apo-Tf in Tris (2-amino-2-hydroxymethylpropane-1,3-diol)-HCl buffer, the percentage of the total (54)MnCl(2) bound to Tf was approximately 85% and 10%, respectively. One hour after intravenous (iv) injection of pH 8.6 buffer-treated (54)MnCl(2) and untreated (54)MnCl(2), both tracers were concentrated similarly in the choroid plexus in the ventricles and distributed in other brain regions. Six days after iv injection, both pH 8.6 buffer-treated (54)MnCl(2) and untreated (54)MnCl(2) tracers were concentrated in the superior olivary complex, inferior colliculi, and red nuclei, although the former radioactivity was lower than the latter. Moreover, Tf-bound (54)Mn was prepared and injected iv into rats. The radioactivity from Tf-bound (54)Mn, which was also concentrated in the same regions, e.g., the superior olivary complex, was the lowest of all three traces. Tf-bound (54)Mn was stable during incubation with serum for 1 hr. It is likely that more Mn is transported into the brain when Mn is not bound to Tf. When Tf bound (54)Mn and (54)MnCl(2) were unilaterally injected into the lateral ventricle, radioactivity was distributed only around the ipsilateral ventricle in the Tf-bound (54)Mn group 7 days after injection, whereas it was distributed more extensively in the (54)MnCl(2) group. It is likely that Tf-bound Mn in the CSF is less readily transported into the brain parenchymal cells than the non-Tf-bound form. These results suggest that Mn is transported into the brain efficiently via a Tf-independent uptake system. PMID- 10679795 TI - Muscular dystrophy in adult and aged anti-NGF transgenic mice resembles an inclusion body myopathy. AB - The role of nerve growth factor (NGF) and its receptors in the physiology of skeletal muscles has not been extensively studied in animal models. We describe the production of transgenic lines of mice expressing a neutralizing antibody against NGF (alphaD11) and the morphological and histochemical analysis of skeletal muscles from adult and aged anti-NGF mice. This study reveals that the chronic deprivation of NGF results in a decreased size of myofibers of dorsal and hindlimb muscles in adult but not in postnatal day (P)2 mice. In myofibers from adult anti-NGF mice, the presence of central nuclei, vacuolization of the cytoplasm, and inflammatory cell infiltration was observed. The immunohistochemical analysis of these muscular fibers revealed an upregulation of p75 expression, a decrease in adenosine triphosphatase (ATP)ase activity, and a subsarcolemmal Congo Red-positive staining. Immunostaining with an antibody against amyloid precursor protein showed an increased labeling of the cytoplasm of myofibers from adult and aged anti-NGF mice. These features are reminiscent of human myopathies, such as inclusion body myositis. We conclude that NGF deficits might be relevant for a class of human myopathies. PMID- 10679796 TI - Localization of glyoxylate dehydrogenase and glyoxylate-complex molecules in the rat prefrontal cortex: enzymohistochemical and immunocytochemical study. AB - Glyoxylic acid is synthesized and catabolized in cells of vertebrates; several pathways have been described. In previous papers, we have demonstrated the localization in some areas of the rat cerebral cortex both of beta-NAD-dependent glyoxylate dehydrogenase (glyoDH), using an enzymohistochemical method, and of glyoxylate-complex molecules, using immunocytochemical procedures. In this study we have applied these two techniques in various areas of the prefrontal cortex with different histological cytoarchitecture. GlyoDH has been located in most neurons, in some glial cells, and in capillary wall structures in all cortical layers of all areas of the rat prefrontal cortex. Antibodies against glyoxylate complex molecules showed positive immunoreactivity in scattered neurons, mostly of multipolar or stellate appearance, from layers III, IV, and V in the medial precentral area, but not in cortical areas 24, 25, or 32 of the prefrontal cortex. Immunoreaction was found in the periphery of neuronal perikarya and in some of their processes. These results demonstrate the existence of a particular area-dependent neuronal cortical system, of specific but uncertain function, related to glyoxylic acid and/or glyoxylate compounds. At the electron microscope level, positive reaction was associated with synaptic sites, axonal filaments, glial cells, and several components of the blood-brain barrier. These localizations suggest the involvement of glyoxylate derivatives in synaptic functioning and also in glial cell functions. PMID- 10679797 TI - Membrane asymmetry and DNA degradation: functionally distinct determinants of neuronal programmed cell death. AB - The ability to elucidate the molecular mechanisms that modulate programmed cell death (PCD) may provide the crucial clues to unravel the cellular basis of neurodegenerative disorders. Employing both a novel assay to follow serially PCD in individual living neurons and the neuroprotective agent lubeluzole as an investigative tool, we examined the development of nitric oxide (NO)-induced PCD over time through the reversible annexin V labelling of membrane phosphatidylserine (PS) exposure and the electron microscopy of genomic DNA in primary rat hippocampal neurons. Exposure to the NO generators SNP (300 microM) or NOC-9 (300 microM) alone increased annexin V-positive neurons in the population from 7% +/- 4% in untreated cultures to 13% +/- 4% at 1 hr and to 61% +/- 5% at 24 hr. Administration of a neuroprotective concentration of lubeluzole (750 nM) at the time of NO exposure initially prevented the exposure of PS residues, but consistently maintained DNA integrity over a 24 hr period. During posttreatment paradigms of lubeluzole (750 nM) at 2, 4, and 6 hr following NO exposure, progression of membrane PS inversion was reversed and subsequently suppressed over a 24 hr course. Our work illustrates that neuronal PCD is composed of at least two physiologically distinct and separate pathways that consist of the externalization of membrane PS residues and the independent maintenance of genomic DNA integrity. In addition, neuronal injury is fluid and reversible in nature, suggesting a "window of opportunity" for the repair and reversal of neurons yet to be committed to PCD. PMID- 10679798 TI - Peripheral nerve regeneration and cholesterol reutilization are normal in the low density lipoprotein receptor knockout mouse. AB - Following peripheral nerve injury, cholesterol from degenerating myelin is retained locally within macrophages and subsequently reutilized by Schwann cells for synthesis of new myelin during nerve regeneration. Substantial evidence indicates this conservation and reutilization of cholesterol is accomplished via lipoprotein-mediated intercellular transport, although the identities of the lipoproteins and their receptors are unresolved. Because Schwann cells in regenerating nerve are reported to express the low-density lipoprotein (LDL) receptor (LDLR), we used the LDLR knockout mouse to examine the potential role of this receptor in cholesterol reutilization. Sciatic nerves were crushed in knockout and wild-type mice and examined 3 days to 10 weeks later. Morphometric analyses and measures of mRNA levels for myelin protein P(0), indicate that axon regeneration and myelination proceed normally in the LDLR knockout mouse. We therefore measured hydroxy-methylglutaryl-coenzyme A (HMG-CoA) reductase activity and mRNA levels to determine whether Schwann cells compensated for the absence of the LDLR by upregulating cholesterol synthesis. Unexpectedly, these measures remained at the same downregulated levels found in regenerating nerves of wild type animals. The apparently normal nerve regeneration, coupled with the lack of any compensatory upregulation of cholesterol synthesis in the LDLR knockout mice, indicates that other lipoprotein receptors must be primarily involved in cholesterol uptake by Schwann cells. PMID- 10679799 TI - Activated recombinant human coagulation factor VII (rFVIIa) therapy for abdominal bleeding in patients with inhibitory antibodies to factor VIII. AB - Eight patients with inhibitors to factor VIII (4 hemophilia A and 4 nonhemophilic) were treated with recombinant activated factor VII (rFVIIa) to control severe abdominal bleeding. The recombinant factor was supplied under an open-label, emergency-use program to patients previously unresponsive to one or more alternative therapies. Therapy with rFVIIa was administered for nine separate bleeding events; one patient was treated for two separate bleeding episodes. Patients were treated for an average of 9 days and received a mean total dose of 5.2 mg of rFVIIa for control of bleeding. Treatment was considered successful and hemostasis adequate in 7 of the 9 episodes (78%). Treatment with rFVIIa was partially successful in one other episode. Four patients in this series experienced serious adverse events; all the adverse events were considered unrelated to rFVIIa therapy. The results of this limited series indicate that rFVIIa is an effective means of managing life-threatening abdominal bleeding in individuals with hemophilia or acquired antibodies to factor VIII. PMID- 10679800 TI - High-level, stable expression of blood group antigens in a heterologous system. AB - The detection and identification of blood group antibodies in patients is crucial for successful allogeneic blood transfusions. Current methods are highly subjective and rely on red blood cells (RBCs), which simultaneously express many blood group antigens, have a short shelf-life, and carry potential biohazard risks. To overcome these problems, we have used the approach of expressing individual blood group antigen-bearing proteins in a heterologous system. We report here the high-level surface expression of type I (Knops), type II (Kell), and type III/multi-pass (Duffy) membrane proteins that carry blood group antigens in mouse erythroleukaemic (MEL) cells using a vector containing the beta-globin locus control region. Importantly, the antigens expressed were detected specifically by a panel of patients' sera containing alloantibodies at sensitivities that are comparable to antigen-positive RBCs. Furthermore, in contrast to other mammalian expression systems, antigen expression was stable following freezing and thawing of the cell lines. Thus, this system has the potential both to replace the current use of RBCs by providing a one step method to detect and identify blood group antibodies and to allow the automation of antibody identification for the clinical laboratory. PMID- 10679801 TI - Cyclophosphamide, etoposide, vincristine, adriamycin, and dexamethasone (CEVAD) regimen in refractory multiple myeloma: an International Oncology Study Group (IOSG) phase II protocol. AB - A 4- day continuous intravenous (CIV) infusion of vincristine and doxorubicin with high-dose dexamethasone (VAD) regimen is a standard refractory multiple myeloma (MM) regimen. A Phase II study of a CEVAD regimen, i.e., VAD plus etoposide administered as a 96-hr continuous infusion, was carried out with IV bolus cyclophosphamide. Thirty-six patients were treated on study and received a total of 114 cycles of CEVAD: median 2 cycles (range 1-8). No patient achieved a CR. The overall rate of PR was 15/36 (42%). Patients achieved maximal response after a median of 4 (range 3-6) courses. PR rates were 40% (4/10) in patients with primary refractory disease, 48% (11/23) in patients with secondary refractory disease, 31% (6/19) in patients who had failed previous VAD therapy, and 50% (7/14) in patients receiving 2nd or subsequent relapse therapy. Three patients died during their initial cycle of therapy from rapidly progressive disease and sepsis. Overall median survival was 24 weeks with a 1-year survival of 33.3% ?95% confidence interval of 20-46%?. Myelosuppression was the most frequent adverse event with NCI grade 2 neutropenia and/or thrombocytopenia in 15% of first cycles, grade 3 in 20%, and grade 4 in 65%. Two-thirds of patients had at least one episode of grade 3 or 4 sepsis. In 15% of septic episodes positive blood cultures were obtained. Overt cardiotoxicity was seen in two patients. CEVAD as used in this study was not more effective than VAD in terms of overall response rate or survival. PMID- 10679802 TI - Strong inverse correlation between serum TPO level and platelet count in essential thrombocythemia. AB - Serum thrombopoietin (TPO) levels in 50 essential thrombocythemia (ET) patients were measured using a highly sensitive sandwich ELISA. In nine cases, TPO levels were measured at two points with different platelet counts. ET patients showed significantly higher serum TPO levels (n = 59, 2.70 +/- 2.74 fmol/mL, P < 0.0001) than those of normal individuals (n = 29, 0.83 +/- 0.36 fmol/mL). Twenty-three previously untreated ET patients also showed significantly higher serum TPO levels (1.33 +/- 0.75 fmol/mL, P = 0.0066) than normal individuals. Extremely high serum TPO levels (5.46 +/- 3.68 fmol/mL) were observed in ET patients with normal platelet counts. Furthermore, a strong inverse correlation was found between serum TPO levels and platelet counts in ET patients (R = -0.729, P < 0. 0001). This inverse correlation also held for each of nine cases with two-point TPO measurements. In the clinical course of ET, megakaryocyte mass may parallel the platelet mass before and after chemotherapy. Although it is unknown whether overproduction of TPO exists or not in ET, total platelet and megakaryocyte mass, i.e., the total number of c-Mpl, may play a role to regulate serum TPO levels. PMID- 10679803 TI - Study on complex formation between recombinant human thrombomodulin fragment and thrombin using surface plasmon resonance. AB - Human thrombomodulin (hTM) is a newly described endothelial cell associated protein that functions as a potent natural anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. The affinity constant of recombinant human soluble TM (rhs-TM) and a peptide containing active site of hTM fragment (f-hTM) with thrombin were determined using the surface plasmon resonance. The interaction of f-hTM with thrombin could be analyzed by a simple model, whereas the association and the dissociation steps of rhs-TM with thrombin consisted of at least two kinds of interaction phases. The dissociation constant for complex (K(D)) of f-hTM and thrombin was determined to be 205 nM, which was more than twice as high as that of rhs-TM (6.7 and 75 nM). The lower affinity of f-hTM was not due to the slow association rate but to the rapid dissociation rate. It comes clear that f-hTM interacts with thrombin rapidly. PMID- 10679804 TI - Ticlopidine-induced aplastic anemia: development of chromosomal abnormalities and response to immunosuppressive therapy. AB - Severe aplastic anemia is a well-recognized complication of ticlopidine therapy that carries a high mortality. Therapy with colony-stimulating factors or corticosteroids has been largely ineffective in this disorder. We report a case of ticlopidine-induced aplastic anemia that was successfully treated with cyclosporine and high-dose dexamethasone. The patient rapidly responded to immunosuppressive therapy and had a normal hemogram after cessation of immunosuppression. On long-term follow-up, the patient developed a progressive macrocytic anemia. Repeat bone marrow evaluation demonstrated myelodysplasia with erythroid hypoplasia. An associated chromosomal abnormality consisting of a t(3;16) (q21; p13.3) translocation was detected. This is the first report of a chromosomal abnormality associated with ticlopidine induced marrow aplastic anemia. PMID- 10679805 TI - Antibody studies of factor VIII inhibitor in a case with Waldenstrom's macroglobulinemia. AB - We report a case of Waldenstrom's macroglobulinemia with prominent bleeding tendency; laboratory investigation revealed an elevated activated partial thromboplastin time. Further laboratory evaluation showed circulating factor VIII anticoagulant, deemed polyclonal IgG, with a titer of 700 Bethesda Units/ml. The factor VIII inactivation kinetics of the patient plasma were identical to those of a type II inhibitor, and the inhibitor was found to recognize the A2 domain of the factor VIII heavy chain. Apparently, paraprotein is not always the cause of reduced activity of coagulation factors in neoplastic dysproteinemias. PMID- 10679806 TI - Fibrinogen Longmont: a dysfibrinogenemia that causes prolonged clot-based test results only when using an optical detection method. AB - A new fibrinogen variant was discovered as a result of discrepancies found in routine laboratory screening. The patient, a healthy 37-year-old woman, had a mild bleeding history. Initial coagulation studies on the patient revealed a prolonged prothrombin time (PT) and a normal activated partial thromboplastin time (APTT). Further investigation on the patient and her mother demonstrated both had a PT with no end point using an optical detection method (ACL3000+) and a normal PT using an electromechanical detection method (ST4 Clot Detection System). The APTT for both the patient and her mother were essentially normal with both optical and mechanical detection methods. The patient and her mother also had markedly prolonged thrombin time and reptilase time results on the ACL3000+, but they were normal on the ST4. Coagulation test results on the patient's father were all normal. We believe the fibrinogen defect in this family may affect fibrin polymerization only enough to effect light scatter interpretation, while there is enough polymerization to increase plasma viscosity and yield an end point using an electromechanical analyzer. This report should alert pathologists and clinicians to possible discrepancies between mechanical and spectrophotometric clot testing methods. PMID- 10679807 TI - The cost of treating immune thrombocytopenic purpura using intravenous Rh immune globulin versus intravenous immune globulin. AB - Multiple factors, including efficacy, toxicity and cost, may influence the decision to treat immune thrombocytopenic purpura (ITP) with intravenous immune globulin (IVIG) or intravenous Rho (D) immune globulin (IV RhIG). We conducted a survey of 50 hospitals in 31 states to determine the costs for treating ITP using conventional doses for IVIG or IV RhIG, based on package insert recommendations. The average cost for a dose of IVIG ($2,771) was 71.7% ($1,157) more than that for a dose of IV RhIG ($1,614). In the absence of clearly defined differences in clinical outcomes when treating ITP with IVIG or IV RhIG, the difference in cost may be an important factor in selecting the treatment. PMID- 10679808 TI - Rapid detection of glucose-6-phosphate dehydrogenase type A-(202A/376G) deficiency by allele-specific polymerase chain reaction. PMID- 10679809 TI - Severe hemolytic anemia following high-dose intravenous immunoglobulin administration in a patient with Kawasaki disease. PMID- 10679810 TI - Philadelphia chromosome positive acute lymphocytic leukemia arising from aplastic anemia. PMID- 10679811 TI - Molecular characterization and chromosomal localization of mouse Puralpha gene. AB - Puralpha is a 39-kDa sequence-specific single-stranded DNA/RNA binding protein with the ability to modulate transcription of several genes containing the Pur element in their promoter region. Human and mouse Puralpha exhibit an extraordinary degree of conservation with only two changes at amino acid residues 49 and 306. A 15-kb genomic clone encompassing the mouse Puralpha gene was isolated by screening the mouse genomic library, using a PCR-amplified fragment from human Puralpha cDNA. Results from sequencing analysis confirmed the isolated genomic clone to be Puralpha and not the other members of the Pur family, including Purbeta. Characterization of the mouse Puralpha gene by restriction analysis/Southern blotting and sequencing revealed that the Puralpha gene contains only one intron within the 5' UTR and the open reading frame was intact. Using chromosomal markers, the Puralpha gene was mapped to chromosome 18 in mouse and rat. PMID- 10679812 TI - Induction of apoptosis in HL-60 cells by N(6)-benzyladenosine. AB - Treatment of HL-60 cells with micromolar concentrations of N(6)-benzyladenosine (N(6)-benzylaminopurine riboside [BAPR]) led to the occurrence of apoptosis in a concentration-dependent manner. Incubation period as short as 2 h in the presence of BAPR was sufficient for triggering irreversible changes leading to apoptosis even after the transfer of cells to the standard medium (without BAPR). Cell death induced by BAPR proceeded rapidly and in a very synchronous fashion. Detailed study of temporal changes in a chromatin structure and DNA integrity showed that the movement of chromatin toward the nuclear periphery is the fundamental event within dying cells. We demonstrated that this rearrangement of chromatin is irreversible and it takes place without apparent DNA degradation. The extensive DNA cleavage seems to be a rather late event, as it was observed in cells that exhibited a typical apoptotic morphology (apoptotic bodies). On the basis of temporal changes in the ATP level within dying cells, it is concluded that ATP is essential for the movement of chromatin toward the nuclear envelope but not for the subsequent chromatin condensation leading to the formation of apoptotic bodies. DNA fragmentation also seems to be ATP independent. BAPR interfered with neither pyrimidine nor purine biosynthesis, as none of the tested bases and the corresponding nucleosides prevented or reduced apoptosis in BAPR treated cells. Adenosine was the only exception that substantially reduced the effect of BAPR. Since transport of exogenous adenosine into cells was essential to manifest its protective effect, we assume that adenosine is a competitive inhibitor of adenosine kinase and thus reduces intracellular phosphorylation of BAPR. Indeed, 4-amino-3-iodo-1(beta-D-ribofuranosyl)pyrazolo[3,4-d]pyrimidine, a potent inhibitor of adenosine kinase, fully prevented BAPR-induced apoptosis. These results suggest that cytotoxic effect of BAPR is related to its activation by phosphorylation within cells, rather than to its interaction with extracellular adenosine receptors. PMID- 10679813 TI - Dietary regulation and localization of apoptosis cascade proteins in the colonic crypt. AB - This study was designed primarily to assess the localization of apoptosis cascade proteins along the rat colonic crypt and secondarily to test whether the activity and/or localization of these proteins are affected by the enrichment of the diet with the soluble fiber pectin. Expression of apoptosis cascade proteins was assessed in isolated colonocytes harvested from the luminal and basal crypt colonocyte populations. Two different dietary regimens were tested: a standard diet (diet A), and a diet enriched in pectin (diet B), a soluble fiber that undergoes fermentation in the cecum and produces high concentrations of intracolonic short-chain fatty acids. Caspase-1 expression was maximal in luminal colonocytes of rats fed diet B, as evidenced by Western blot and immunohistological analyses. Expression of the cleaved poly(ADP-ribose) polymerase product was elevated in both the luminal and basal colonocytes of the pectin-fed group, whereas in rats fed diet A, the expression was lower, especially in basal crypt colonocytes. The highest expression of the antiapoptotic protein Bcl-2 was observed in the lower compartments of the colonic crypt tissue and was maximal in the rat group fed a standard diet. The apoptotic index in colonocytes of rats fed diet B was higher than that measured in rats fed diet A. Cumulatively, our results indicate that apoptosis cascade proteins are differentially localized along the lumen-crypt axis, and their expression and activity may be controlled by dietary components. These results may, at least partially, account for the documented protective effect of butyrogenic fibers on colorectal cancer. PMID- 10679814 TI - Modified intranuclear organization of regulatory factors in human acute leukemias: reversal after treatment. AB - Acute leukemias arise secondary to chromosomal aberrations that cause dysfunctions in gene regulation and regulatory factors. Significant differences in morphology between acute leukemic and nonleukemic hematopoietic cells are readily observed. How morphologic changes of the nuclei of acute leukemic cells relate to the underlying functional alterations of gene expression is minimally understood. Spatial modifications in the representation and/or organization of regulatory factors may be functionally linked to perturbations of gene expression in acute leukemic cells. Using in situ immunofluorescence microscopy, we addressed the interrelationships of modifications in nuclear morphology with the intranuclear distribution of leukemia-related regulatory factors (including ALL 1, PML, and AF-9) in cells from patients with acute leukemia. We compared the localization of leukemia-associated proteins with various factors involved in gene transcription and RNA processing (e.g., RNA polymerase II and SC-35). Our findings suggest that there are leukemia-associated aberrations in mechanisms that direct regulatory factors to sites within the nucleus. This misplacement of key cognate factors may contribute to perturbations in gene expression characteristic of leukemias. PMID- 10679815 TI - Stimulant-free preculture in heterologous serum-supplemented medium induces unresponsiveness of T cells to subsequent mitogenic stimulation. AB - Quite often freshly isolated lymphocytes are kept in culture before experimentation for 1 or more days without any stimulus. Most of the time, culture is supplemented with fetal bovine serum (FBS) which is heterologous to all species except bovine. In the present study, we found that freshly isolated murine T cells show a good proliferative response to concanavalin A (Con A) and phorbol ester (PMA)/ionomycin in FBS medium, without any detectable background proliferation. However, the cells kept in the same culture without any stimulus for prolonged period of time (referred to as preculture in this report) showed reduced response to Con A and PMA/ionomycin in a time-dependent manner. Almost a complete loss of response to Con A was observed within 1 day of preculture. However, loss of response to PMA/ionomycin was observed only after 2 days of preculture. Interestingly, similar preculture in autologous mouse serum supplemented media did not cause any loss of the response to these mitogens. The loss of responsiveness of T cells during preculture in heterologous serum was irreversible. The heterologous serum-induced unresponsiveness of T cells to these mitogens was also prevented by adding Calphostin C, a specific protein kinase C (PKC) inhibitor, during preculture in heterologous serum. These results showed that prolonged stimulant-free preculture in heterologous serum induces irreversible unresponsiveness of T cells to mitogens through the down regulation of T cell receptor signaling pathway, which can be prevented by autologous serum or a PKC inhibitor. PMID- 10679816 TI - Transcriptional control elements and complex initiation pattern of the TATA-less bidirectional human thymidylate synthase promoter. AB - The nucleotide sequences that are important for transcription of the human thymidylate synthase gene were analyzed by deletion and site-directed mutagenesis of the promoter region. Deletion analyses from the 5' and 3' ends indicated the presence of multiple positive and negative elements. The promoter had approximately the same strength in the normal or inverted orientation. The region between 161 and 141 nt upstream of the translational start codon was found to be both necessary and sufficient for high-level promoter activity in both directions and was designated the essential promoter region. This region, which is highly conserved in human, mouse and rat TS promoters, contains potential binding sites for Ets, Sp1, and LSF transcription factors. Site directed mutagenesis of each of these elements led to large decreases in promoter strength. However, inactivation of potential Sp1 and E2F elements adjacent to the essential promoter region led to increases in promoter strength. The transcriptional start site pattern was analyzed by S1 nuclease protection assays of mRNA isolated from cells transiently transfected with TS minigenes. Multiple start sites were detected, most of which were between 160 and 120 nt upstream of the AUG codon. PMID- 10679817 TI - Interaction of HIV-1 Tat with Puralpha in nuclei of human glial cells: characterization of RNA-mediated protein-protein binding. AB - A complex between the Tat protein, encoded by human immunodeficiency virus type 1 (HIV-1), and the cellular protein, Puralpha, has been implicated in activation of the late promoter of JC virus (JCV) and in enhancement of JCV DNA replication. JCV is the causative agent of progressive multifocal leukoencephalopathy (PML), an acquired immunodeficiency syndrome (AIDS) opportunistic infection of the brain. Puralpha also binds the HIV-1 TAR RNA element and activates HIV-1 transcription, suggesting a role for RNA binding in the action of this protein. Using immunoelectron microscopy, we find that in human glial cells expressing both proteins, Tat and Puralpha are colocalized in extranucleolar chromatin structural elements. The colocalized Puralpha and Tat are nearly exclusively nuclear, although individual proteins can be seen in both nucleus and cytoplasm, suggesting a preferential tropism of the complex for the nucleus. Analysis of the interaction between purified proteins indicates that the Tat-Puralpha interaction is strongly enhanced by the presence of RNA. Tat amino acids from 37-48 are essential for Tat binding. Residues 49-72, including the TAR RNA-binding domain, are critical for binding to Puralpha, while Cys(22), in the Tat transactivation domain, is responsible for an important global effect. Puralpha repeat II domains are involved in the interaction, and a polypeptide based on one such sequence inhibits binding. After RNase treatment of Puralpha enhancement of Tat binding can be partially restored by addition of a single-stranded JCV DNA PUR element, to which Tat does not bind. The results indicate that the Tat-Puralpha interaction is direct, rather than through an RNA link, and that RNA binding configures Puralpha for optimal interaction with Tat. PMID- 10679818 TI - Cyclin C is a primary 1alpha,25-dihydroxyvitamin D(3) responding gene. AB - 1alpha,25-dihydroxyvitamin D(3) (VD) is a pleiotropic nuclear hormone that also has effects on cell cycle regulation. VD and its synthetic analogues are known inhibitors of cellular growth and inducers of apoptosis, however, the primary mediator genes of these effects largely remain unknown. In order to identify novel targets for VD, that may be involved in the regulation of the cell cycle, a differential display PCR (ddPCR) approach was applied to the MCF-7 human breast cancer cell line, which provided the gene for cyclin C as an interesting candidate. Quantitative assessment of cyclin C expression showed that the gene was significantly upregulated by VD and its analogues, EB1089 and CB1093 both on the level of mRNA expression and more so on the level of protein expression in MCF-7 cells. Upregulation of cyclin C protein expression could also be confirmed in MeWo human melanoma and in U937 human promyelocytic leukemia cells. This observation adds a new gene candidate to the list of primary VD responding genes. Cyclin C is not a typical cyclin, as it apparently modulates the activity of the RNA polymerase II complex, which provides fresh insight into the mechanisms of cell cycle and general transcriptional regulation by VD and its analogues. PMID- 10679819 TI - Multiple low-dose and single high-dose treatments with streptozotocin do not generate nitric oxide. AB - Streptozotocin (STZ) is a widely used diabetogenic agent that damages pancreatic islet beta cells by activating immune mechanisms, when given in multiple low doses, and by alkylating DNA, when given at a single high dose. Actually, STZ contains a nitroso moiety. Incubation of rat islets with this compound has been found to generate nitrite; moreover, photoinduced NO production from STZ has been demonstrated. These reports have suggested that direct NO generation may be a mechanism for STZ toxicity in diabetogenesis. Several other studies have denied such a mechanism of action. This study has shown that (1) the multiple low-dose (MLDS) treatment does not stimulate NO production at the islet level; in fact, nitrite + nitrate levels and aconitase activity (also in the presence of an NO synthase inhibitor, namely NAME) remain unmodified; RT-PCR analysis demonstrates that this treatment does not stimulate iNOS activity; (2) the high-dose (HDS) treatment does not stimulate NO production; in fact nitrite + nitrate levels remain unmodified and iNOS mRNA levels are not altered, although aconitase activity is significantly decreased. Moreover, we have confirmed that the MLDS treatment is able to decrease SOD activity by day 11 and that STZ, given in a single high dose, transiently increases superoxide dismutase (SOD) values (24 h from the administration), then dramatically lowers SOD levels. On the basis of our results, we conclude that STZ, "in vivo" is unable to generate NO, both as a MLDS or HDS treatment, thus excluding that NO exerts a role in streptozotocin dependent diabetes mellitus. PMID- 10679820 TI - Isolation and characterization of the chicken vitamin D receptor gene and its promoter. AB - The sequences from several independent cDNA clones encoding the chicken vitamin D receptor as well as primer extension assay have clearly delineated the 5' terminus and the transcriptional start site. Screening a chicken genomic library produced genomic clones containing vitamin D receptor (VDR) gene fragments. Restriction map of clone 8 showed that the 18.6-kb chicken VDR fragment has exons 1 and 2, intron 1, part of intron 2, and 7-kb 5' flanking region. Exons 1, 2, and 3 found in the chicken VDR gene shares low homology with its mammalian counterparts (i.e., E1A, E1B, and E1C in human). By contrast, the fourth exon and following exons for the coding region of VDR gene are highly conserved between avian and mammalian species. While the fourth exon bears the ATG sites for translation initiation in mammals, the third exon in birds has two extra ATG sites for leaky translation as determined previously. Thus, the avian VDR has more N-terminal sequence than the mammalian VDR and is found in two distinct forms. The 5' flanking region from genomic clone 8 shares considerable homology in several regions with the human and mouse VDR promoters. Moreover, the 5' flanking region of chicken VDR gene possesses promoter activity, as shown by its ability to drive the luciferase reporter gene in cell transfection assays. Like other steroid receptor promoters, the chicken VDR promoter contains no TATA box but possesses several GC boxes or SP1 sites. A series of deletional promoter constructs established that the proximal GC boxes are the major drivers of gene transcription, while the more upstream sequences have repressive elements. PMID- 10679821 TI - Cloning the full-length cDNA for rat connective tissue growth factor: implications for skeletal development. AB - The mammalian osteopetroses represent a pathogenetically diverse group of skeletal disorders characterized by excess bone mass resulting from reduced osteoclastic bone resorption. Abnormalities involving osteoblast function and skeletal development have also been reported in many forms of the disease. In this study, we used the rat mutation, osteopetrosis (op), to examine differences in skeletal gene expression between op mutants and their normal littermates. RNA isolated from calvaria and long bones was used as a template for mRNA differential display. Sequence information for one of the many cDNA that were selectively expressed in either normal or mutant bone suggested that it is the rat homologue of connective tissue growth factor (CTGF) previously cloned in the human, mouse, and other species. A consensus sequence was assembled from overlapping 5'-RACE clones and used to confirm the rat CTGF cDNA protein coding region. Northern blot analysis confirmed that this message was highly (8- to 10 fold) over-expressed in op versus normal bone; it was also upregulated in op kidney but none of the other tissues (brain, liver, spleen, thymus) examined. In primary rat osteoblast cultures, the CTGF message exhibits a temporal pattern of expression dependent on their state of differentiation. Furthermore, CTGF expression is regulated by prostaglandin E(2), a factor known to modulate osteoblast differentiation. Since members of the CTGF family regulate the expression of specific genes, such as collagen and fibronectin, we propose that CTGF may play a previously unreported role in normal skeletal modeling/remodeling. Its dramatic over-expression in the op mutant skeleton may be secondary to the uncoupling of bone resorption and bone formation resulting in dysregulation of osteoblast gene expression and function. PMID- 10679822 TI - A deficit in collagenase activity contributes to impaired migration of aged microvascular endothelial cells. AB - Angiogenesis is impaired in aging. Delayed neovascularization is due, in part, to slowed endothelial cell migration. Migration requires an optimal level of adhesion to matrix proteins, a process mediated by matrix-degrading metalloproteases (MMPs) such as MMP1. To determine whether impaired angiogenesis in aging is associated with altered synthesis and activity of MMP1, we examined the expression of collagenase and tissue inhibitor of metalloprotease 1 (TIMP1) by immunostain of angiogenic sponge implants from young and aged mice. To characterize the relevance of MMP activity during the movement of aged endothelial cells, the secretion of MMP1 and TIMP1 by late-passage human microvascular endothelial cells (hmEC aged in vitro) and their non-aged (young) counterparts was quantified. The migration of aged human microvascular endothelial cells and the effect of inhibition of TIMP1 on the migration of aged hmEC or collagen I was also measured. Relative to young mice, granulation tissue from aged mice showed less expression of collagenase and increased expression of TIMP1. In vitro, aged hmEC were deficient in MMP1 secretion (55 +/- 13% relative to young cells) and activity but showed increased expression of TIMP1 (280 +/- 109% relative to young cells). Aged hmEC migrated significantly less distance than did young hmEC over a 5-day period (59 +/- 8% relative to young cells). In the presence of a blocking antibody to TIMP1, aged hmEC showed a significant increase in the distance migrated on collagen I over a 5 day period (142 +/- 11% relative to untreated aged hmEC). We propose that deficient MMP1 activity contributes to impaired cellular movement in aged microvascular endothelial cells and that perturbations that enhance collagenase activity increase their migratory ability and angiogenic potential. PMID- 10679823 TI - Regulation of heat shock protein message in Jurkat cells cultured under serum starved and gravity-altered conditions. AB - Although our understanding of effects of space flight on human physiology has advanced significantly over the past four decades, the potential contribution of stress at the cellular and gene regulation level is not characterized. The objective of this ground-based study was to evaluate stress gene regulation in cells exposed to altered gravity and environmentally suboptimal conditions. We designed primers to detect message for both the constitutive and inducible forms of the heat shock protein, HSP-70. Applying the reverse transcriptase-polymerase chain reaction (RT-PCR), we probed for HSP-70 message in human acute T-cell leukemia cells, Jurkat, subjected to three types of environmental stressors: (1) altered gravity achieved by centrifugation (hypergravity) and randomization of the gravity vector in rotating bioreactors, (2) serum starvation by culture in medium containing 0.05% serum, and (3) temperature elevation (42 degrees C). Temperature elevation, as the positive control, significantly increased HSP-70 message, while centrifugation and culture in rotating bioreactors did not upregulate heat shock gene expression. We found a fourfold increase in heat shock message in serum-starved cells. Message for the housekeeping genes, actin and cyclophilin, were constant and comparable to unstressed controls for all treatments. We conclude that gravitational perturbations incurred by centrifugal forces, exceeding those characteristic of a Space Shuttle launch (3g), and culture in rotating bioreactors do not upregulate HSP-70 gene expression. In addition, we found RT-PCR useful for evaluating stress in cultured cells. PMID- 10679824 TI - Transgenic mice with a mutated collagen promoter display normal response during bleomycin-induced fibrosis and possess neurological abnormalities. AB - We have previously identified a potential TGF-beta activation element (TAE) in the rat collagen alpha1(I) promoter at -1624 upstream of the transcriptional start site [Ritzenthaler et al., 1991, 1993]. To determine the importance of the TAE in vivo, we produced transgenic mice carrying 3.6 kb of the rat collagen alpha1(I) promoter linked to the reporter gene chloramphenicol acetyl transferase with and without site-directed mutations that eliminate DNA-protein binding at the TAE site. Tissue-specific expression of the reporter gene in transgenic mice with the mutated collagen promoter was similar to that of transgenic mice with the normal promoter in two genetic backgrounds as judged by in situ hybridization, reporter assays, and immunochemistry. Endotracheal instillation of bleomycin induces lung fibrosis, mediated in part by TGF-beta. Earlier studies indicated that expression of wild-type collagen-reporter gene was upregulated in transgenic mice lungs in response to endotracheal instillation of bleomycin. A similar level of reporter gene upregulation was observed in transgenic mice carrying the mutation in the TAE. Two lines of transgenic mice carrying the mutated promoter construct displayed unexpected neurological abnormalities. In the FVB genetic background, there was a higher than normal incidence of mortality, spontaneous seizures, and an inability to nurture offspring. Histological evidence demonstrated clear abnormalities, including disorderly arrangement of neurons in the hippocampus and significant laminar cortical necrosis in the cerebrum in animals after seizures. In the C57Bl/6 background, there was a high incidence of severe communicating hydrocephalus, early runting, and increased mortality similar to that in transgenic animals with astroglial overexpression of TGF-beta. These animals provide an interesting model system to investigate molecular mechanisms responsible for seizures and hydrocephalus. PMID- 10679825 TI - Parathyroid hormone inhibits collagen synthesis and the activity of rat col1a1 transgenes mainly by a cAMP-mediated pathway in mouse calvariae. AB - We examined the effect of parathyroid hormone and various signaling molecules on collagen synthesis and chloramphenicol acetyltransferase activity in cultured transgenic mouse calvariae carrying fusion genes of the rat Col1a1 promoter and the chloramphenicol acetyltransferase reporter. After 48 h of culture, parathyroid hormone, forskolin, dibutyryl cAMP, 8-bromo cAMP, and phorbol myristate acetate inhibited transgene activity, while the calcium ionophore ionomycin had no effect. Pretreatment of calvariae with the phosphodiesterase inhibitor isobutylmethylxanthine potentiated the inhibitory effect of 1 nM parathyroid hormone on transgene activity and collagen synthesis. Parathyroid hormone further inhibited transgene activity and collagen synthesis in the presence of phorbol myristate acetate. Parathyroid hormone inhibition of transgene activity and collagen synthesis was not affected by indomethacin or interleukin-6. After 48 h of culture, parathyroid hormone inhibited chloramphenicol acetyltransferase activity by 50-85% in cultured calvariae carrying transgenes having progressive 5' upstream deletions of promoter DNA down to -1683 bp. These data show that the inhibitory effect of parathyroid hormone on Col1a1 expression in mouse calvariae is mediated mainly by the cAMP signaling pathway. Prostaglandins and IL-6 are not local mediators of the parathyroid hormone response in this model. Finally, regions of the Col1a1 promoter downstream of -1683 bp are sufficient for parathyroid hormone inhibition of the Col1a1 promoter. PMID- 10679826 TI - Expression and regulation of the cGMP-binding, cGMP-specific phosphodiesterase (PDE5) in human colonic epithelial cells: role in the induction of cellular refractoriness to the heat-stable enterotoxin peptide. AB - Stable toxin (ST) peptides are the causative agents for a severe form of watery diarrhea. These peptides bind to a membrane-associated form of guanylyl cyclase, guanylyl cyclase C. The result is an accumulation of cyclic guanosine monophosphate (cGMP) in the intestinal cell, regulating protein kinase activity and the phosphorylation of a number of proteins involved in ion transport across the intestine. Using the human T84 colonic cell line as a model system, we show that cGMP accumulation in these cells after ST application is regulated by the activity of the cGMP-binding, cGMP-specific phosphodiesterase (PDE5). The presence of human PDE5 in this cell line was confirmed by Western blot analysis, using an antibody raised to the bovine enzyme, and by the observation that cGMP hydrolytic activity detected in T84 cell lysates was almost completely inhibited by low concentrations of zaprinast, a specific inhibitor of PDE5. An increase in activity of PDE5 was observed in T84 cell lysates on exposure to the ST peptide and prolonged exposure of T84 cells to the ST peptide led to the induction of cellular refractoriness in these cells, which was largely contributed in terms of an increased rate of degradation of cGMP in desensitized cells as a result of PDE5 activation. This activation was correlated with an increase in the affinity of the enzyme for the substrate cGMP, as well as an increased affinity for zaprinast. We provide evidence for the first time that cGMP levels in the human colonocyte are regulated by the cGMP-hydrolytic activity of PDE5 and suggest that the expression and regulation of PDE5 in the intestine could therefore be important in controlling cGMP-mediated signaling in this tissue. PMID- 10679827 TI - Keeping score. PMID- 10679828 TI - Standardisation of ambulatory urodynamic monitoring: Report of the Standardisation Sub-Committee of the International Continence Society for Ambulatory Urodynamic Studies. PMID- 10679829 TI - A simplified urinary incontinence score for the evaluation of treatment outcomes. AB - There are no standardized definitions for anti-incontinence therapy outcomes. The present study was conducted to evaluate whether the incorporation of several non invasive outcome measures into a new score may serve as a meaningful outcome instrument. Ninety-four consecutive sphincteric incontinent women who underwent a pubovaginal sling by a single surgeon were enrolled. All patients underwent a full clinical evaluation, including pre- and post-operative questionnaires, 24 hour voiding diary, and 24-hour pad test. Surgery outcomes were classified twice: First, by analyzing the patient questionnaire, voiding diary, and pad test separately, according to previously published criteria, and second, by combining the three outcome tools into a new response score. The new score was constructed in a simple, easy-to-remember format and divided into five categories: cure, good response, fair response, poor response, and failure. All patients were evaluated at least 1 year post-operatively. Comparison of the old and new classifications suggests that the new response score provides a more accurate evaluation of the surgical outcomes. Although 64 to 69% of the patients were originally classified as cure according to the old classification, only 44.7% were re-classified as cure by the strict criteria employed in the new score. Furthermore, the response score also differentiates between various degrees of clinical improvement (i.e., good, fair, or poor response). Twenty-five (26.6%) patients, most of whom were previously classified as cure, were re-classified as good response, whereas 20 others were re-classified as fair (13. 9%), or poor (7.4%) response. Seven (7.4%) patients were re-classified as surgical failures. All were diagnosed pre operatively as having complex sphincteric incontinence. Specific failure rates were therefore 11.3% for complex and 0% for simple cases. In conclusion, the suggested post-operative response score incorporates in a user-friendly format three popular outcome tools (i.e., 24-hour diary, 24-hour pad test, and patient questionnaire) and seems to reflect the surgical results more accurately. Further studies are needed to assess its validity and reproducibility in other treatment modalities. Neurourol Urodynam. 19:127-135, 2000. PMID- 10679830 TI - A severity index for epidemiological surveys of female urinary incontinence: comparison with 48-hour pad-weighing tests. AB - In epidemiological surveys of female urinary incontinence, it is not feasible to demonstrate urine loss objectively. The aim of this study was to develop a valid epidemiological instrument (a severity index) for assessing the severity of incontinence. The severity index is based on information about frequency (four levels) and amount of leakage (two or three levels). By multiplication, an index value (1-8 or 1-12) is reached. This index value is further categorized into a severity index of three or four levels. The index was compared with the results of 315 pad-weighing tests performed by 265 women in hospital and general practice. Data from an epidemiological survey were also re-analyzed by applying the four-level severity index. Mean pad-weighing results (grams per 24 hours, 95% confidence interval) for the three-level severity index was slight (6; 2-9), moderate (17; 13-22), and severe (56; 44-67). For the four-level severity index, the results were slight (6; 2-9), moderate (23; 15-30), severe (52; 38-65), and very severe (122; 84-159). Spearman's correlation coefficient for pad-weighing results and the three-level severity index was 0.47 (P < 0.01) and for the four level severity index 0.54 (P< 0.01). The four-level severity index gave a more balanced distribution among the women in the clinical materials, and data from the epidemiological survey showed that the four-level severity index identifies a sub-group of older women with very severe incontinence. The four-level severity index seems to be a valid representation of incontinence severity as measured by pad-weighing tests in women presenting for clinical care. It should be considered a potentially valid measure of incontinence severity in epidemiological studies. Neurourol. Urodynam. 19:137-145, 2000. PMID- 10679831 TI - Effects of menstrual cycle and urinary tract instrumentation on uroflowmetry. AB - We sought to compare bladder emptying function in normal women during the proliferative and secretory phases of the menstrual cycle and to evaluate whether urethral catheterization affected uroflowmetry parameters. Forty-nine normal volunteers (ages 19-42 years) were recruited and underwent uroflowmetry in the proliferative and secretory phases of the menstrual cycle. A serum progesterone level of <3.0 ng/mL defined the proliferative phase. During the proliferative phase, volunteers underwent uroflowmetry analysis when a sensation of fullness was appreciated. A post-void residual volume was determined, and the bladder was filled with 400 mL of normal saline. Repeat uroflowmetry analysis was then performed. This two-step procedure was repeated at a separate visit during the secretory phase. Voided volume, residual volume, maximum and average flow rates, time-to-maximum flow, and duration of flow were recorded. Wilcoxon signed-rank tests were used for statistical analysis. A two-tailed alpha value of <0.05 defined statistical significance. Our analysis was limited to the 33 patients whose predicted menstrual dating correlated with the obtained progesterone levels. During the follicular phase, we found significantly faster maximum (P < 0.0001) and average flow rates (P = 0.01), along with a shorter time-to-maximum flow (P < 0.0001) and shorter duration of flow (P < 0.0001), during the pre catheterized void than the post-catheterized void. Similar results were observed in the secretory phase with the exception of a slightly higher residual volume (P = 0.05). No difference was seen in any measured uroflowmetry parameter when comparing similar voids between phases of the menstrual cycle. We conclude that when evaluating pre-menopausal patients, uroflowmetry may be scheduled and performed during either phase of the menstrual cycle. Neurourol. Urodynam. 19:147 152. PMID- 10679832 TI - A mathematical micturition model to restore simple flow recordings in healthy and symptomatic individuals and enhance uroflow interpretation. AB - We describe a model and report a new method to extract quantified data from the simple analysis of whole uroflow curves in healthy and symptomatic individuals. Recorded flow curves were compared with the curves theoretically predicted from a mathematical micturition model. This model was developed by relating each physiological event occurring during micturition to a set of mathematical equations. Due to improvements in speed of computer calculations, a very fast and adaptable mathematical micturition model became available. A total of 302 uroflow curves from 142 patients (61 men and 81 women) were studied. The control group consisted of seven men and 25 women; the symptomatic groups comprised 54 men and 56 women. The mathematical model was applied to analyze all the recorded curves. For patients with lower urinary tract symptoms, specific modelization parameters were introduced according to the clinical condition to be tested. Using two compulsory (patient sex and voided volume) and two optional parameters (clinical condition and urethral catheter size), our mathematical model was able to produce uroflow calculated curves similar to observed curves. In the control group, the calculated and recorded uroflow curves were found to superimpose with an impressive accuracy, i.e., a quadratic error <1%. Test-re-test studies gave the same determination of the specific parameters. In benign prostatic hyperplasia patients, the compressive effect on both prostatic urethra and bladder neck was separately identified. The same intra-individual values were found at 2-week intervals in the group with no treatment (P = no significance), whereas after 3 months of treatment with an alpha-blocker, a decrease in values was noted in responder patients (P < 0.001). In women with various degrees of cystocele, a constrictive effect was identified and found to be identical for successive flows during the same urodynamic testing. This large prospective study demonstrated the relevance of a sophisticated, heavily computerized micturition model, taking into account physiological voiding parameters, to the study of flow in healthy individuals and patients with benign prostatic hyperplasia or cystocele. Curve fitting led to the determination of critical events during flow such as break point and plateau phase. Determination of these events may enhance uroflow interpretation by providing additional information on the detrusor function. Neurourol. Urodynam. 19:153-176, 2000. PMID- 10679833 TI - Editorial comment. PMID- 10679834 TI - Voiding pattern in healthy newborns. AB - A 4-hour observation period has been used in infants to investigate suspected bladder dysfunction. The aim of the present study was to extend the usefulness of this protocol by establishing reference values for voiding frequency, intervals, volumes, and residual urine in healthy newborns. The study included 51 healthy newborns, 26 girls and 25 boys, aged 3 to 14 days. During a 4-hour period, all micturitions and residuals were recorded as well as feeding, sleeping, crying, and defecations. The observation was completed with the child undressed to observe the urinary stream during one void. Different provocation tests were tried to induce urinary leakage. All newborns voided with a stream, about once per hour, with a median volume of 23 mL. For each voiding parameter, there was a large inter- and intra-individual variability. Double voidings were common as well as sizable residual volumes. The diuresis was about six times higher than in healthy school children. The healthy newborns did not leak during provocation tests such as manual compression of the bladder. Neurourol. Urodynam. 19:177-184, 2000. PMID- 10679835 TI - The distribution of vesicular acetylcholine transporter in the human male genitourinary organs and its co-localization with neuropeptide Y and nitric oxide synthase. AB - Because doubt still remains concerning the distribution of nerves that are unequivocally cholinergic in the human genitourinary organs, we have used a specific marker, namely, an antibody to vesicular acetylcholine transporter (VAChT), to immunolabel cholinergic axons and cell bodies in specimens of urinary bladder, seminal vesicle, vas deferens, and prostate gland obtained from neonates and children post mortem. In addition some sections were double-immunolabeled with VAChT and either neuropeptide Y (NPY) or nitric oxide synthase (NOS). The results demonstrated a rich cholinergic innervation to the muscle coat of the bladder body with a much less prominent, but nonetheless significant, cholinergic innervation to the smooth muscle components of the seminal vesicle, vas deferens, and prostate. Small ganglia were scattered throughout the detrusor muscle of the urinary bladder, approximately 75% of the intramural neurons being VAChT immunoreactive, whereas approximately 95% contained NPY and approximately 40% contained NOS. VAChT immunoreactivity was observed in 40% of neurons in ganglia scattered throughout the pelvic plexus. Almost all these cholinergic neurons contained NPY and approximately 65% contained NOS. Almost all the cholinergic nerve fibers throughout the genitourinary organs also contained NPY. Although NOS was sparse in the cholinergic nerves of the bladder body, it occurred in the majority of cholinergic nerves at the bladder neck and was also present in a proportion of the cholinergic nerves in the other organs examined. VAChT immunoreactive nerves were also observed in a sub-epithelial location in all the organs examined, the majority containing NPY, whereas a small proportion contained NOS. Although doubt remains about the function of sub-epithelial cholinergic nerves in the urinary bladder, the majority of similar nerves in the seminal vesicle, vas deferens, and prostate gland are considered to be secretomotor. Collectively these findings demonstrate that the cholinergic innervation of the male genitourinary system is well established in the neonate and child. Neurourol. Urodynam. 19:185-194, 2000. PMID- 10679836 TI - Acute increase in blood flow to the rat bladder subsequent to partial bladder outlet obstruction. AB - Partial obstruction of the rat bladder outlet initiates a multi-step process during which the bladder progressively loses its functional ability. The first step in this progression is bladder hypertrophy; the organ dramatically increases in size and weight to compensate for the effects of obstruction. Unoperated female rats, age-matched, sham-obstructed rats, and rats that received a partial bladder outlet obstruction were studied. During the first 24 hours after partial bladder outlet obstruction, relative bladder blood flow was measured using a fluorescent microsphere infusion technique and laser Doppler imaging. Nitric oxide synthase (NOS) activities of control and obstructed rat bladder tissues were determined using an enzymatic assay that measures the conversion of (3)H-L arginine to (3)H-L-citrulline. Using the microsphere infusion technique, a significant increase in blood flow to the obstructed rat bladder was observed during the first 24 hours after partial bladder outlet obstruction. Relative bladder blood flow increased approximately sixfold at 4 and 6 hours post obstruction and remained elevated through 24 hours of obstruction. Sham operations (evaluated after 6 hours after surgery) resulted in a minor increase in blood flow that did not reach statistical significance. Relative blood flow to the spleen, measured in the same rats, was not significantly changed. Laser Doppler measurements also identified a significant increase in rat bladder blood flow after outlet obstruction and showed that increased blood flow could be detected as early as 1 hour post-obstruction. Interestingly, despite the significant differences in bladder blood flow between control and early post obstructed rat bladders, NOS activities of control and obstructed rat bladders were comparable. The increase in bladder blood flow precedes the urothelial, fibroblast and smooth muscle cell hyperplasia, and the smooth muscle hypertrophy that occurs after obstruction. We propose that, in response to surgical induction of partial outlet obstruction, acute up-regulation of bladder blood flow may be an initiating factor for subsequent bladder cell proliferation and smooth muscle hypertrophy. Neurourol. Urodynam. 19:195-208, 2000. PMID- 10679837 TI - Editorial comment. PMID- 10679838 TI - Authors' reply. PMID- 10679839 TI - Stress urinary incontinence: prevalence among nulliparous compared with primiparous and grand multiparous pre-menopausal women. PMID- 10679840 TI - Delusions and hallucinations in Alzheimer's disease: prevalence and clinical correlates. AB - OBJECTIVES: The purpose of this study was to examine the frequency of delusions and hallucinations in patients with Alzheimer's disease (AD) and to investigate factors associated with each or the combination of the two. DESIGN: This was a cross-sectional, case-control study. SETTING: Neuropsychiatry and Memory Group, The Johns Hopkins University, USA. PARTICIPANTS: Three hundred and forty-two community-residing patients with probable AD according to NINCDS/ADRDA criteria were included in the study. MEASURES: Patients were assessed clinically for the presence of psychotic symptoms using the DSM-IV glossary definitions. The patients were also rated on standardized measures of cognitive impairment, depression, extrapyramidal symptoms, functional impairment and general health. RESULTS: Seventy-five (22%) AD patients had delusions only, nine (3%) had hallucinations only and 30 (9%) had both delusions and hallucinations. Hallucinations were associated with less education, African-American race, more severe dementia, longer duration of illness, falls and use of anxiolytics. Delusions were associated with older age, depression, aggression, poor general health and use of antihypertensives. Patients with both delusions and hallucinations were similar to the patients with delusions only. CONCLUSIONS: This study confirms the high prevalence of psychotic symptoms in AD patients encountered in clinical practice and suggests that individual psychotic symptoms have different associations. PMID- 10679841 TI - Is aggressive behaviour influenced by the use of a behaviour rating scale in patients in a psychogeriatric nursing home? AB - OBJECTIVE: To study the influence of the introduction of a behaviour rating scale on reported incidence and management of aggressive behaviours in patients in a psychogeriatric nursing home. DESIGN: An 8-month prospective cohort intervention study. SETTING: Two wards of a Dutch psychogeriatric nursing home with residents of 65 years and older. PATIENTS: All residents of the two wards were included (N=75). INTERVENTION: Social Dysfunction and Aggression Scale (SDAS) assessed at weekly intervals during 4 months after a 4 months' baseline period. ASSESSMENTS: During 8 months at 2 months' interval the BOP (ie the validated Dutch version of the Stockton Geriatric Rating Scale); frequency of aggressive behaviour reported at daily nursing staff report; mean prescriptions of psychotropic and somatic drugs; number of days a patient was submitted to physically restrictive measures. RESULTS: Eleven patients (N=11) did not complete the study; information on 64 patients was analysed. The frequency of aggressive behaviour reported by the nursing staff increased, while prescriptions of psychotropic drugs decreased. No alteration was found for BOP scores, mean prescriptions of somatic drugs and the use of physically restrictive measures. CONCLUSIONS: The introduction of a behaviour rating scale does influence the reported incidence and management of aggressive behaviour. Prospective intervention studies should include a stabilization phase for measurements prior to any planned trial. PMID- 10679843 TI - Do personality traits predict the occurrence of Alzheimer's disease? AB - OBJECTIVE: To identify specific premorbid personality traits in patients with Alzheimer's disease (AD). DESIGN: A prospective case-control study. SETTING: A memory clinic of a department of geriatric medicine in a teaching hospital. PATIENTS: Fifty-six consecutive patients with probable AD. Sixty-five controls with Parkinson's disease (PD). MEASURES: Premorbid personality traits were assessed using the relative rating version of the Munich Personality Test (MPT). RESULTS: The AD patients showed higher neuroticism than the controls with PD (p=0.013). In comparison with MPT normative values for psychiatric inpatients, the AD patients scored significantly (p<0.05) lower on neuroticism and higher on frustration tolerance and rigidity. CONCLUSION: Our results support the assumption of specific premorbid characteristics in AD patients, ie increased neuroticism and rigidity. More research is needed to confirm the existence of typical premorbid personality traits in AD. PMID- 10679842 TI - Factors predicting the relapse of depression in old age. AB - BACKGROUND: Studies in mixed-aged populations show differences between the predictors of a relapse and those of a long-term course of depression, supporting the hypothesis about similar differences among the aged. AIM: The aim was to identify the factors predicting or related to a relapse of depression among the Finnish elderly having recovered from depression during treatment. MATERIAL AND METHODS: The population consisted of 70 depressed (DSM-III criteria) elderly (60 yr-) Finns having recovered from depression during treatment as determined 15 months after baseline. By the 4-year follow-up after the recovery, 20 patients had relapsed and 50 persons were non-depressed. RESULTS: The logistic regression model showed major depression and psychomotor retardation to be independent predictors. Relapses were not related to stressors in life or physical illnesses occurring during the follow-up. CONCLUSIONS: Major depressive elderly patients have a high risk for relapses without the occurrence of the stressors or physical illnesses. In clinical practice, major depressive elderly patients should be followed up in order to detect and treat potential relapses as early as possible. Cooperation between psychiatrists and general practitioners is needed in the follow-up. Theoretically, the results suggest the assumption of a biochemical aetiology of major depression. PMID- 10679844 TI - A pilot study of sibling resemblance in later life. AB - OBJECTIVE: Behavioural genetic studies of later life are strictly limited. We carried out a community-based pilot study of sibling resemblance with the primary aims of establishing the feasibility of such work in this population and estimating genetic influence on depression and its risk factors. METHODOLOGY: Data were collected on surviving siblings of individuals interviewed in previous phases of an epidemiological study of the elderly (the Gospel Oak survey); scales relevant to the investigation of late life depression and its risk factors were utilized. Since families tend to be geographically scattered, the interview was conducted by telephone. Comparisons were made between data relating to the siblings and those obtained on the probands. Variability in phenotypic traits and environmental measures was partitioned into between- and within-family variation, in order to distinguish familial and non-familial sources of variation. Intraclass correlations were used to estimate the strength of genetic influences on continuous measures, while pairwise concordances were calculated for dichotomous traits. RESULTS: Thirty-two siblings from 20 families were ultimately identified and interviewed. Intraclass correlations for the Depression and Dementia Diagnostic Scales and the Handicap Scale were 0, 0.27 and 0.22, respectively. Those for number of life events, number of friends in contact and number of neighbours in contact were 0.08, 0.03 and 0, respectively. Concordance for both depression caseness and dementia caseness was 0. DISCUSSION: There were difficulties carrying out this study, which are discussed. The study is the first of its kind to examine familial resemblance for the common disorders of old age. Establishing ways of engaging elderly families with research will be a challenge that future research will need to meet. PMID- 10679845 TI - Development of simple cognitive function measures in a community dwelling population of elderly in Spain. AB - OBJECTIVE: To develop and assess the consistency and validity of simple cognitive function measures for an elderly population with low levels of formal education for use in a longitudinal study of dementia. METHODS: Data were from the population longitudinal study 'Growing old in Leganes' (Spain). In 1993, a random sample of 1540 people over 65 was drawn from the City Roll of Leganes from which 1284 (83%) were successfully interviewed. Measures of memory and orientation were based on the SPMSQ (Short Portable Mental Status Questionnaire), the Barcelona test and the short story from EPESE (Established Populations Epidemiologic Studies of the Elderly). Non-response to a test item was coded as an error. Internal consistency was assessed by factor analysis and Cronbach's alpha. Construct validity was examined with multiple linear regressions of the proposed measurements on variables chosen from the existing literature on cognitive function. RESULTS: Two factors, memory and orientation, emerged from the factor analysis. Internal consistency of the proposed indexes for memory and orientation was acceptable. Memory and orientation scores were summed into one summary index of cognition. Associations between covariates and both cognitive indexes were in the expected direction. Among those highly functional, orientation was influenced by illiteracy due to higher error rates in the time orientation items based on dates; however, memory and summary scores were not significantly different by literacy status. A large proportion of the variance in IADL was explained by the memory and orientation measures. CONCLUSION: The memory and orientation indexes are valid and reliable measurements of cognitive function for use in a population of community dwelling elderly with low levels of formal education and high rates of illiteracy. PMID- 10679846 TI - The effects of emotion-oriented approaches in the care for persons suffering from dementia: a review of the literature. AB - OBJECTIVE: This article presents an overview of the results of intervention studies in various emotion-oriented approaches in the care for people suffering from dementia. Recommendations are made with regard to clinical practice and future research. DATA SOURCES: We searched for references (1990-99) in several bibliographical databases, i.e. Medline, PsycLit, Embase, Sociofile and Current Contents. The terms 'dementia' and 'Alzheimer's disease' were linked separately to the search terms: emotion-oriented, validation (therapy), sensory integration/sensory stimulation/snoezelen, simulated presence therapy and reminiscence (therapy)/life-review. Based on references in the articles found, other publications were traced. STUDY SELECTION: We started from the 'emotion oriented' approaches used in 24-hour care distinguished by the American Psychiatric Association (1997) i.e. validation, sensory stimulation/integration, simulated presence therapy and reminiscence. We selected research articles that describe intervention, design, measuring instruments and results. DATA EXTRACTION: The articles were analyzed with regard to research group, setting, design, effect variables, intervention, measuring instruments, statistical analyses and results. DATA SYNTHESIS: It is shown that mainly positive results (including increased social interaction and decrease of behavior problems) are achieved with these emotion-oriented approaches. Unfortunately many studies have methodological limitations and are done independently, which makes comparison difficult. CONCLUSIONS: Despite the limited cogency of the studies we traced, the results are promising. Emotion-oriented care approaches offer the opportunity to tailor the care to the individual needs of dementing elderly and can be complemented with other psychosocial approaches (e.g. psychomotor therapy and music therapy) when necessary. The challenge for the care sector is to develop guidelines to determine which approach should be applied to whom and when. Scientific research can contribute by examining which emotion-oriented approaches, possibly in combination with each other or with psychosocial therapies, effect an increase in the well-being and improve functioning in which patients. PMID- 10679847 TI - Suicidal ideation in acutely medically ill elderly inpatients: prevalence, correlates and longitudinal stability. AB - BACKGROUND: Suicidal ideation among acutely medically ill elderly inpatients has been sparsely studied. A prospective study measuring the prevalence, correlates and longitudinal stability of suicidal ideation in acutely medically ill elderly inpatients was undertaken. METHOD: Suicidal ideation was measured using the Beck Scale for Suicidal Ideation (BSSI) and the items of pessimism, life not worth living and a wish to die on the Brief Assessment Schedule (BAS). Formal measures of physical illness, functional disability and handicap were also used. Patients were seen at the outset and at about 6 months. RESULTS: The prevalence of suicidal ideation on the BSSI and the BAS items of pessimism, life not worth living and a wish to die were 36%, 60%, 33% and 22%, respectively. These four variables were significantly inter-correlated. The BSSI was significantly associated with BAS depression scores (P=0.0001), BAS depression caseness (P=0.0001) and prescription of antidepressants (P=0.007). Similar results were ascertained for the BAS items of pessimism, life not worth living and a wish to die. CONCLUSIONS: Further studies examining the longitudinal stability of suicidal ideation coupled with intervention studies to reduce suicidal ideation are required. PMID- 10679848 TI - Can chronic neuroleptic treatment promote sleep disturbances in elderly schizophrenic patients? AB - It has been proposed that sleep disturbances, especially reduced delta sleep, are related to a poor outcome in schizophrenia. To determine whether long-term treatment with neuroleptics can promote sleep disturbances by increasing the risk of a nocturnal myoclonus syndrome (NMS) (=periodic movements in sleep) related insomnia, we performed all-night polysomnography in 10 chronically ill schizophrenic patients who had been under neuroleptic therapy for a mean of 27 years. NMS-related insomnia was detected in all 10 patients. Potential pathophysiological relationships between long-term neuroleptic therapy and NMS occurrence are discussed. Our findings suggest that long-term administration of neuroleptics favours the appearance of insomnia. PMID- 10679849 TI - Sertraline in the treatment of minor depression in nursing home residents: a pilot study. AB - 'Minor' depression affects up to 50% of residents in long-term care facilities and is associated with considerable discomfort, disability and risk of morbidity. Despite the prevalence of this problem, few studies addressing the treatment of these patients have been conducted. In an open clinical trial, 12 nursing home residents who met the DSM-IV description for minor depressive disorder were treated with sertraline for 6 weeks. Adverse effects and clinical response were monitored. All residents tolerated their medication without any significant side effects. At the completion of the study, the Hamilton Depression Rating Scale and Global Assessment Scale change scores both indicated significant improvement and 75% of the residents met criteria for 'remission'. This preliminary study provides evidence that nursing home residents with minor depression tolerated treatment with sertraline and improved clinically. PMID- 10679850 TI - Quality of life in dementia patients in long-term care. AB - OBJECTIVE: To evaluate variables associated with quality of life (QOL) in dementia residents in a long-term care facility using a recently standardized and validated dementia-specific QOL scale (ADRQL). METHOD: A cross-sectional, case control design was employed using validated scales to assess dementia-related symptomatology. Thirty-two facility staff members were interviewed to assess the QOL of 120 patients meeting DSM-IV for dementia criteria residing in long-term care. RESULTS: ADRQL scores were higher in assisted living residents than in skilled nursing facility residents. In univariate analyses, worse orientation, greater physical dependency, depression, and treatment with anxiolytics were associated with lower ADRQL scores. In multivariate analyses, lower scores were associated with worse orientation, greater physical dependency, depression, and anxiolytic treatment. CONCLUSIONS: Residents exhibited better QOL than expected. Future longitudinal studies should address if reorientation, activity therapy, treatment of depression, and avoidance of benzodiazepines might improve QOL in this population. Interventions that might improve orientation and physical abilities, such as cholinomimetic therapies, psychosocial interventions, or behavioral strategies, should also be studied in future research on QOL. PMID- 10679851 TI - Anastomosis between the external branch of the superior laryngeal nerve and the recurrent laryngeal nerve. AB - An incidental finding in the anatomy lab showed up a plexus of the external branch of the right superior laryngeal nerve (SLN), including an anastomosis with the recurrent laryngeal nerve (RLN). The external branch of the SLN divided in two extensions: The ventral extension reached the mesopharynx laterally and by supplying the latter, ended at the cricothyroid muscle. The dorsal extension formed a plexus a finger's breadth beneath the inferior margin of the pharynx, on the lateral aspect of the esophagus. The anastomosis ran from the lower part of the plexus to the RLN along the esophagus, laterally. PMID- 10679852 TI - Radiological anatomy of the intratemporal course of facial nerve. AB - Preoperative evaluation of the facial nerve (FN) anatomy within the temporal bone by high-resolution computed tomography (HRCT) helps in minimizing surgical trauma to the nerve. In order to demonstrate the radiological correlation of the intratemporal FN, eight adult, formalin-preserved cadavers were studied by comparing the transaxial and coronal sections of HRCT with anatomic microdissection findings. It was possible to visualize all segments of the FN canal in its intratemporal course. The most difficult part of the FN to demonstrate was the pyramidal section. Anatomic microdissection findings were consistent with the HRCT images. It was concluded that adequate information on the FN anatomy could be obtained from standard HRCT scans. PMID- 10679853 TI - Cadaver profile at university of Stellenbosch Medical School, South Africa, 1956 1996. AB - Data on 1,698 cadavers donated during the period 1956-1996 were obtained from files of the Department of Anatomy and Histology at the University of Stellenbosch Medical School, Tygerberg Hospital, South Africa, to project a profile of the characteristics of those accepted on the program for dissection. A breakdown of the data also provided information on the profile of donors belonging to different population groups. Donors to our program were predominantly male (68%) and predominantly colored, which in South Africa identifies those of mixed heritage (63%). The average age of death was 55 years (range 15-98). Donors belonging to the white population group had the highest female : male ratio. Circulatory disorders accounted for most deaths in the white population group (48%), whereas cancer was the leading cause of death in the colored and black population groups ( approximately 25%). Pulmonary tuberculosis accounted for 13% deaths in the colored population group, 14% of deaths in the black population group, but only 0.5% of deaths in the white population group. Cervical cancer and breast cancer accounted for approximately one-third of cancer deaths in women, with cervical cancer more common in colored and black female donors and breast cancer more common in white female donors. The cadaver profile in general reflects the health status of the different population groups in South Africa. The profiles of the colored and black groups reflect that of disadvantaged population groups (a high prevalence of infectious disease; relatively young populations), whereas the white donor profile is that of a privileged population group (a high prevalence of degenerative disease; aging population). PMID- 10679854 TI - Anatomical knowledge and clinical evaluation of the muscles of mastication. AB - In this survey, we identify the positive role that an instructional anatomy course has prior to a workshop on temporomandibular dysfunction. Not only did it result in enhanced clinical evaluation of the muscles of mastication acquired by the participants, but it also approximated the skills of dental and medical practitioners. PMID- 10679855 TI - Theoretical and analytical embryology of conjoined twins: part II: adjustments to union. AB - A previous report in this journal (Spencer, 2000) discussed the probable embryologic etiology of conjoined twins, along with a system of classification based on the embryological structures postulated to be involved in the union. Part II correlates and compares the variations in the abnormal development of the individual organ systems in more than 1,200 actual cases, revealing details of embryogenesis not considered in previous publications. The site, incidence, and range of anomalies in the conjoined structures, as well as the associated malformations, follow a definite pattern as the union proceeds from one area to another; many can be explained in relation to the proposed embryologic adjustments to union, including both temporal and spatial influences. In addition, six currently inexplicable or unclassifiable cases are briefly described (including one with 12 feet), as well as two examples of early abnormal conjoined twins. PMID- 10679856 TI - Position, shape, and dimension of the maxilla in unoperated cleft lip and palate patients: review of the literature. AB - The inhibition of growth and development resulting from surgical treatment of the cleft lip and palate is a widely discussed topic. Various studies have been conducted in search of answers as to how the untreated upper jaw develops, focusing on individuals with untreated cleft lip and palate as found in so-called Third World countries. This study offers the opportunity to compile literature dealing with the research and description of untreated unilateral cleft lip and palate. The focus was to take a closer look at groups of individuals with complete unilateral cleft lip and palate, who had received no surgical treatment at all, as well as groups who had received surgical treatment of only the cleft lip. The upper jaw of untreated cleft lip and palate patients most often adopts a protruded position without enlarging the maxilla itself. The horizontal dimension tends to be reduced, whereas the vertical dimension is normal. The upper jaw of patients with unilateral cleft lip and palate who received surgical treatment of the lip more often adopted a retruded position. The model analysis showed no clear-cut tendencies. There seemed to be a degree of regional variation. Considering the relatively small number of recruitable individuals with untreated cleft lip and palate, the introduction of a standard method of evaluation is desirable. This would significantly facilitate the comparison of different studies with each other in the future. The first steps in this direction have already been initiated. PMID- 10679857 TI - Aberrant left gastric vein directly draining into the liver. AB - An aberrant left gastric vein found in dissection is reported here. The right gastric vein did not exist and only the left gastric vein originating from the lesser curvature of the stomach was present. It directly entered the liver without typically joining the trunk of the portal vein. After giving off a small branch to the liver parenchyma, the left gastric vein merged into the left branch of the portal vein. This aberrant left gastric vein may correspond to the phylogenetic and ontogenetic "left portal vein." The aberrant left gastric vein is considered to play an important role as a portal collateral pathway of the portal system, which is critical not only in anatomy but also in clinical diagnosis. PMID- 10679858 TI - Sternalis muscle: topic for debate. PMID- 10679859 TI - The new anatomy: A forecast of hope PMID- 10679860 TI - Standards in health care and medical education. PMID- 10679861 TI - What is anatomy? Implications for anatomy as a discipline and for Clinical Anatomy as a journal. AB - Anatomy as a discipline is explored against a background of the growth of scientific knowledge and developments within modern universities. The structural overtones of anatomic approaches provide intellectual cohesion for the approaches of complementary disciplines. When anatomy departments function in a transdisciplinary manner, they encompass a range of areas from molecular biology to functional anatomy and biological anthropology. Suggestions are made for a journal, such as Clinical Anatomy, to provide stimulus for the further development of anatomy as a coherent discipline. PMID- 10679862 TI - Preface PMID- 10679863 TI - A framework for assessing health-related quality of life among children with cancer. PMID- 10679864 TI - Commentary on assessing health-related quality of life in children with cancer. PMID- 10679865 TI - The Miami pediatric quality of life questionnaire: parent scale. AB - Because there were limited measures available to assess health-related quality of life (HRQL) in children with chronic illnesses, this study was initiated to develop an empirically derived questionnaire for use in evaluating HRQL issues in children treated for cancer. Extensive interviews were conducted with 30 families of children with cancer, 10 of pre-school age, 10 of school age and 10 of adolescent age. Responses were videotaped and transcribed, then categorized to develop a pool of 56 items, which were administered to 132 children with cancer and to their parents. This report focuses on parental responses to objective items and ratings of importance of each of these items. Three primary categories, Self-Competence, Emotional Stability and Social Competence, were identified, each of which had solid internal consistency, sensitivity and reliability across 1 month intervals. The measure demonstrated the ability to discriminate between children with different types of cancer, offers an alternative to measures relying on expert judgment to assess HRQL and may lead to greater inclusion of psychological and social concerns as primary factors in determining HRQL in children participating in clinical trials. PMID- 10679866 TI - Assessment of health-related quality of life in acute in-patient settings: use of the BASES instrument in children undergoing bone marrow transplantation. AB - The Behavioral, Affective and Somatic Experiences Scale (BASES) represents a set of tools for assessing aspects of health-related quality of life (HRQL) in patients undergoing active, intensive therapy. Separate versions have been developed for parent, nurse and patient reports. The scales were constructed to be sensitive to change and appropriate for repeated measures in longitudinal designs. We report preliminary results with these measures from a sample of 105 children undergoing bone marrow transplantation (BMT). Adequate reliability of the instruments is documented through measures of both internal consistency and cross-informant consistency. Several analyses provide evidence of the clinical validity of the measures. Repeated-measures ANOVAs indicated reliable patterns of change over time, with trajectories that conformed to a priori predictions. Discriminative validity was demonstrated through detection of significant differences in the predicted direction between patients undergoing allogeneic and autologous BMT. Additional evidence for validity comes from the very similar symptom trajectories in parent, nurse and patient reports. Differences between the BASES and other measures of HRQL are identified and alternative uses of the instruments are discussed. PMID- 10679867 TI - Health status and health-related quality of life in long-term adult survivors of pediatric solid tumors. AB - We have examined the influence of selected factors (gender, marital status, socio economic status, co-morbid conditions, access to medical care, age at diagnosis, intensity of therapy and time since diagnosis) on subsequent health status and health-related quality of life (HRQL) of long-term survivors of pediatric solid tumors. Two hundred and twenty individuals who had survived a pediatric solid tumor 15 years or longer completed telephone and written assessments of their current status. Health status was assessed using the Late Effects of Normal Tissues toxicity scale. HRQL was investigated using the Ferrans and Powers Quality of Life Index-Cancer (QLIC) and the EORTC Quality of Life Questionnaire C30 (QLQ-C30). Results indicated that health status and HRQL were better in survivors treated with low-intensity therapy. One hundred and thirty respondents (59.1%) reported at least 1 serious toxicity. Dyspnea and fatigue were commonly reported in survivors of Hodgkin's disease. Correlational analyses showed that predictors of health status included socio-economic status, marital status and the presence of co-morbid factors. Mean HRQL scores for the 4 domains of the Ferrans and Powers QLIC and the functional scales of the EORTC QLQ-C30 indicated that most of the survivors were experiencing moderately good to excellent HRQL. One-third of survivors reported that their history of cancer had an adverse impact on their current financial status. Prediction models constructed for 3 of the domains from the 2 HRQL instruments are presented (health and functioning, global HRQL and financial impact). Within these 3 models, consistent predictors of HRQL outcomes included health status, presence of dyspnea or pain, marital status and socio-economic status. PMID- 10679868 TI - Assessment of health status and health-related quality of life in survivors of Hodgkin's disease in childhood. AB - Although the great majority of children with Hodgkin's disease survive with modern treatment strategies, the list of late sequelae is long, yet there is no published information on the comprehensive health status and health-related quality of life (HRQL) in this population. In the experience of a single institution, survivors of Hodgkin's disease in childhood were invited to self report on their health status using a 15-item questionnaire connected to the Health Utilities Index, a series of multi-attribute health status classification systems that, in turn, are linked to preference functions which provide single attribute and global utility scores for HRQL. The mean global utility score was 0.85 (on a 0 = dead to 1. 0 = perfect health scale), a figure less than that in survivors of acute lymphoblastic leukemia (ALL) but comparable to that in survivors of brain tumors (0.84) or extremely low birthweight (ELBW 0.82). The burden of morbidity is emphasized by the ratio of the numbers of health states per patient:0.67 for survivors of Hodgkin's disease, 0.66 for survivors of brain tumors, 0.39 for survivors of ELBW, 0.47 for survivors of high-risk ALL and 0.28 for survivors of standard-risk ALL. In Hodgkin's disease survivors, the attributes affected most commonly and severely were pain, cognition and emotion. This experience demands exploration of the health status and HRQL in a much larger cohort of such survivors, perhaps in the context of co-operative group studies. PMID- 10679869 TI - A comparison of parent and adolescent reports describing the health-related quality of life of adolescents treated for cancer. AB - Our objectives were to compare adolescent and parent ratings of the health related quality of life (HRQL) of adolescents treated for cancer, to compare the HRQL of adolescents who were on treatment vs. the HRQL of those who were off treatment following their diagnosis with cancer and to assess the HRQL of adolescents who were at different points of time following their diagnosis with cancer. The HRQL of 70 adolescents (aged 10 to 18 years) consecutively attending the Women's and Children's Hospital Oncology Clinic in South Australia was assessed by means of standard questionnaires. Parents completed the Child Health Questionnaire, the Functional Status II(R) Questionnaire and the Impact-on-Family Scale. Adolescents completed the self-report version of the Child Health Questionnaire. In general, there was good agreement between parent and adolescent reports. However, parents of adolescents receiving active treatment for cancer reported that their illness was having a greater impact on the adolescents' physical functioning than was reported by the adolescents. The psycho-social functioning of adolescents in single-parent families was reported also by parents to be worse than that of adolescents in 2-parent families. The physical functioning of adolescents had only a weak relationship with parental status but a significant relationship with treatment status. Despite generally good agreement between parent and adolescent reports describing the HRQL of adolescents treated for cancer, it cannot be assumed that reports from parents are always an accurate reflection of the views of the adolescents. Studies examining the influence of independent factors on adolescents' HRQL must take into account differences in reports from these 2 informants and the possibility that key independent variables have differing relationships with the various domains which comprise adolescents' HRQL. PMID- 10679870 TI - Health-related quality of life in pediatric bone marrow transplant survivors: according to whom? AB - Historically, health-related quality of life (HRQL) assessment in pediatrics, including the few validated instruments in pediatric oncology, has been based on proxy reporting, relying primarily on parental assessment. Children have been deemed incapable of providing consistent and reliable information about their level of functioning or state of well-being. Previous studies have been hampered by either limited or poor correlation among the proxy reporters, i.e., teachers, parents and physicians, and in comparisons to disease severity. Simply stated, proxy reporters have greater agreement about what the child can do vs. what the child thinks or feels. Comparisons among proxy reporters have been hindered also by a lack of parallel content in the instruments used, which may result in poorly congruent assessments simply because the instruments measure different constructs. In addition to the measurement issues, the emotional milieu of the parent, particularly the mother, has been shown to influence assessments of the child's functioning. Maternal distress, marital adjustment and health locus of control all co-vary with reports of the child's behavior. What, then, is the proxy reporter telling us about the child? We conducted a cross-sectional study of school-aged pediatric bone marrow transplant (BMT) patients at our institution to evaluate children's self-reported HRQL and functional status. We formally tested the Child Health Rating Inventories (CHRIs), a recently developed generic health-status measure, with its companion measure, the Disease Impairment Inventories-Bone Marrow Transplant (DSII-BMT). Separate questionnaires were administered to patients, parents and physicians at a scheduled outpatient visit after BMT. The questionnaires were designed to have parallel content. All responses were confidential. The psychometric properties of the CHRIs and DSII BMT are reported elsewhere. In brief, the responses of all raters were reliable, based on measurements of internal consistency. The children's self-reported health status was correlated significantly with the physicians' disease severity rating (DSR) across all generic and disease-specific domains. In contrast, parental reports of child health status were not correlated significantly with the DSR for disease-specific problems or the child's pain. Parental ratings deviated most from the children's ratings within the dimensions of mental health and quality of life (p < 0.001). For the entire sample, parental ratings were significantly lower than the children's ratings. Within the subgroup "early after transplant (<6 months)", parental ratings were significantly lower than the children's self-reports in all categories. In the subgroup ">12 months after transplant", with the exception of mental health and quality of life, parental scores were the same as or higher than the children's ratings. Our results confirm previous studies that the parental reporting of children's health status is a complex construct and that valuable information can be elicited directly from the children. Further research is needed to substantiate these findings, particularly in longitudinal applications with adequate sample sizes. PMID- 10679871 TI - Somatization, anxiety and depression as measures of health-related quality of life of children/adolescents with cancer. AB - This descriptive study of health-related quality of life of children with cancer compared children/adolescents', parents' and teachers' ratings for somatization, depression and anxiety to determine if there were significant correlations among respondent scores. In addition, the percentage of agreement among respondents and significant differences based on age, gender, use of cranial radiation and treatment status were measured. Forty-three children/adolescents with cancer, currently receiving therapy for at least 1 year or who had completed therapy for no more than 3 years (excluding children who had received bone marrow transplants or who had brain tumors), were recruited, with a parent and teacher, from 3 university medical centers. The Behavioral Assessment System for Children questionnaires for children/adolescents, parents and teachers were used. Parents reported a higher level of depression for the children/adolescents with cancer than did the teachers or the children/adolescents themselves. Parents reported a higher level of anxiety for the children/adolescents than did the teachers. High positive correlations were found among scores from parents and teachers and among scores from parents and children/adolescents for the anxiety and depression but not somatization subscales. Children/adolescents and teachers had high, positively correlated scores only for the depression subscale. High, positive correlations were found between somatization, anxiety and depression within each group of respondents. A significant percentage of agreement between all respondents on ratings for at-risk status was obtained only for the depression subscale. Age was the only variable found to have an influence on scores and only for the anxiety subscale. PMID- 10679872 TI - Health-related quality of life in childhood cancer: discrepancy in parent-child reports. AB - The purpose of our study was to describe reports of parents and of children with cancer on items taken from 4 domains of health-related quality of life (HRQL), bodily pain/distress, general health perceptions, physical functioning and limitations in role/social functioning as a result of physical health, and to examine whether differences in parent-child reports varied as a function of the child's health condition (cancer vs. healthy). Twenty-seven child-parent dyads with cancer and 27 child-parent dyads who were healthy (child ages 8 to 18 inclusive) completed measures of child HRQL [Child Health Questionnaire-Parent Form (CHQ-PF50) and Child Health Questionnaire (CHQ-CF87)] and demographic information at a scheduled out-patient general pediatric or pediatric oncology clinic appointment. Sixteen items included on both the CHQ-CF87 and CHQ-PF50 were examined to compare parent and child reports of child HRQL. As hypothesized, greater discrepancies were evident in the reports of parents of children with cancer than parents of children who are healthy [F(16,31) = 3.98, p < 0.0001]. Statistically significant discrepancies emerged in parent and child responses on 50% of the items in the sample of children with cancer, with parents reporting that their children experience more limitations in their lives than did the children themselves. In the healthy group, statistically significant discrepancies emerged on only 1 of the 16 items (6.3%). PMID- 10679873 TI - Development of the Royal Marsden Hospital paediatric oncology quality of life questionnaire. AB - Our objective was to develop a health-related quality of life measure for use in pediatric oncology. The development process followed the EORTC Quality of Life Study Group (QLSG) guidelines but utilized a parental proxy rating methodology developed within the framework of the EORTC QLSG. Data are reported on the preliminary stages of development, which include interviews in the target population, specialist review of questionnaire content and initial results on the psychometric structure of the measure. The questionnaire has been translated from English to Swedish and Dutch and is available for international field testing. Suggestions for further development of the new measure are described, including the need for parallel forms for use with children and adolescents as well as the parental proxy rating form described here. PMID- 10679874 TI - The Pediatric Cancer Quality of Life Inventory: a modular approach to measuring health-related quality of life in children with cancer. AB - Measurement of pediatric cancer patients' health-related quality of life (HRQL) in phase III randomized, controlled clinical trials is being recognized increasingly as an essential component in evaluating the comprehensive health outcomes of modern anti-neoplastic treatment protocols. Use of a brief core measure of HRQL plus disease-specific symptom modules is a way to assess specific HRQL outcomes with a minimum of subject burden. Demonstrating a measure's feasibility, reliability and validity also represents children's ability to provide reliable and valid responses to HRQL questions. The Pediatric Cancer Quality of Life Inventory (PCQL) Modular Approach consists of a 15-item core measure of HRQL and 2 specific symptom modules: pain and nausea. To validate a patient-report form and a parent-report form, the PCQL was administered to 291 pediatric cancer patients and to their parents. Feasibility and range of measurement, as well as patient-parent concordance, were assessed. Internal consistency reliability was assessed via Cronbach's alpha. Validity was determined by the known-groups approach and by correlating PCQL scores with days missed from school. Patients had minimal missing data, and the range of measurement for the items was good. Patient-parent concordance was large but not perfect. For both patient and parent forms, internal consistency reliability of the PCQL core scale (0.83 and 0. 86, respectively) was strong. The internal consistency reliabilities of the 2 symptom modules for both patient and parent forms were in the acceptable range for group comparisons. Regarding clinical validity, the core scale and the 2 symptom modules distinguished between patients on and off treatment for both patient and parent reports. Further, both patient and parent reports correlated with days of missed school in the past 6 and 12 months. The PCQL Modular Approach has demonstrated acceptable internal consistency reliability and clinical validity for both patient-report and parent report forms. By implication, children are capable of providing reliable and valid responses to these HRQL questions. PMID- 10679875 TI - Standardized quantitative assessment of brain tumor survivors treated within clinical trials in childhood. AB - Important morbidity and impairment of life quality arises from both the primary pathology and therapeutic interventions in children with central nervous system (CNS) tumors. Standardized and systematic collection of morbidity data is a prerequisite of clinical trials in this field. The perception of the survivor is paramount in the determination of quality of life as this variable is dependent on the beholder. Comprehensive assessment of outcome following therapeutic intervention should evaluate this in parallel with other physical and psycho social outcome parameters. A structured, simple schema for the evaluation of survivors of childhood CNS tumors is presented. It is intended to be easily applicable by clinicians within the everyday clinical setting. Information relating to pre- and post-operative states, function, health status and emotional and psychological well-being is collected at regular intervals from diagnosis. Re integration into society and independence are evaluated. Children self-complete health-status assessments where appropriate. Evidence to support this is presented. The schema is intended to provide a basic framework for the monitoring of health status following treatment of CNS tumors in childhood. Regular assessments may identify individuals in need of more detailed investigation and further understanding of the evolution of morbidity in this cohort. Survivors' perception of the impact of documented dysfunction on their health-related quality of life will be determined. Optimization of the planning of future clinical service provision and therapies will result. PMID- 10679876 TI - Studies on health-related quality of life in childhood cancer in the European setting: an overview. AB - Since the beginning of the 1990s there has been a growing interest, in the European setting, in evaluating health-related quality of life (HRQL) in clinical studies. Assessing HRQL in childhood cancer survivors, in particular, is a new field of research. Studies of survivors of leukemia and brain tumors are of special interest since these are the commonest groups of survivors of cancer in childhood. Initial reports suggest that most of the survivors of childhood cancer are in good health with a normal psychosocial status, social life and capacity to cope with activities of daily living. More discriminative evaluations identify a number of subtle problems, such as cognitive deficits in brain tumor survivors or anxiety about a recurrence (especially) in children who have had megatherapy and autologous bone marrow rescue. In this group, pain is also a lasting problem in about one-third of patients. The main problem in these studies is comparability as the study designs vary widely. Little information is available with respect to inter-observer agreement, which is important since it is known that proxy respondents will under- or over-estimate components of the HRQL of the child. The most important base for further development of HRQL research is communication between researchers, in order to exchange experience and to avoid duplication of effort. Communication is necessary also for combining interests and forces in developing a standardized methodology as well as for conducting collaborative studies using equivalent measures. PMID- 10679877 TI - Health-related quality-of-life measures for children. AB - Our purpose is to report the development and psychometric properties of a generic computer-delivered measure of health-related quality of life (HRQL) suitable for children aged 6 to 11 years, the Exeter HRQL scale (EHRQL). The theoretical model adopted is based on a definition of HRQL which assumes that HRQL is the result of discrepancies between an individual's actual self and ideal self. The EHRQL consists of 16 pictures, each of which is rated twice, first in terms of "like me" and second as "I would like to be". The difference between these scores is assumed to be indicative of HRQL. The EHRQL is delivered using a Macintosh Powerbook and takes approximately 20 min. Data are reported for 60 children with asthma (mean age = 8.93 years) and 69 healthy children (mean age = 7.49 years). In addition, children with asthma completed the Childhood Asthma Questionnaire (CAQ) and a measure of self-efficacy. For children with asthma, significant correlations were found between discrepancy scores and 3 of the 4 subscales of the CAQ. In addition, higher discrepancies were found for children with asthma compared with healthy children (p < 0.05). The EHRQL has acceptable internal reliability, and these data provide preliminary support for the theoretical assumption that HRQL reflects perceived discrepancies between an individual's actual self and ideal self. The measure also distinguished, as predicted, between children with asthma and healthy children. Methodological refinements to the EHRQL are suggested. PMID- 10679878 TI - Correlation of the Health Utilities Index Mark 2 cognition scale and neuropsychological functioning among survivors of childhood medulloblastoma. AB - Children surviving medulloblastoma have a high risk for chronic, treatment related neurocognitive deficits. Neuropsychological testing provides important data regarding the comparative toxicities of various therapeutic approaches. However, such testing can be expensive and logistically difficult, especially if a consulting psychologist is not readily available at the treating institution. Our purpose was to investigate the usefulness of a health-related quality of life inventory that does not require a psychologist for completion. We assessed the concurrent validity of traditional intelligence (IQ) testing and levels on the Cognition attribute of the multi-attribute Health Utilities Index Mark 2 (HUI 2) in estimating academic achievement scores of 22 patients treated for medulloblastoma with craniospinal irradiation following surgical resection. The results demonstrated that the Cognition utility scores were significantly lower than scores from the other components of the HUI 2 (Sensation, Mobility, Emotion, Self-Care, Pain). Cognition scores were also significantly positively correlated with IQ and achievement scores. Furthermore, Cognition scores were significantly lower among children who had received special educational services when compared with those who had not received such services. Our results provide preliminary evidence of the potential usefulness of the HUI 2 Cognition attribute in estimating IQ, achievement and the likelihood of the need for special educational services among children treated for medulloblastoma. PMID- 10679879 TI - Mutual concurrent validity of the child health questionnaire and the health utilities index: an exploratory analysis using survivors of childhood cancer. AB - Mutual concurrent validity of 2 generic measures of health-related quality of life (HRQL), the Child Health Questionnaire (CHQ) and the Health Utilities Index Mark 2 (HUI2) and HUI3, was assessed. Data were from 3 centers participating in a Canadian multi-center retrospective cohort study currently in progress to assess psycho-social and physical late effects in children surviving >/=5 years after cancer diagnosis between 1981 and 1990. Exploratory results are from 244 parent reports on HRQL in children <16 years old when studied. Spearman rank-order correlations between sub-scale scores for the CHQ and single-attribute utility scores for the corresponding attribute from the HUI2 and HUI3 were used. As predicted, the correlation between CHQ bodily pain and HUI2 and HUI3 pain was strong, 0.58 and 0.60, respectively, while correlations between CHQ physical functioning and HUI2 mobility and HUI3 ambulation were moderate, both 0.45. Correlations between CHQ mental health and HUI2 and HUI3 emotion were strong, 0.64 and 0.54, respectively, rather than moderate, as predicted. Both the CHQ general health scale and the general health single item were moderately correlated with the HUI2 and HUI3 global utility scores rather than weakly, as predicted (CHQ general health scale and HUI2 and HUI3 global utility were 0.43 and 0.44, respectively; CHQ general health single item and HUI2 and HUI3 global utility were 0. 38 and 0.42, respectively). The CHQ and HUI, which are based on different methodologies (summative Likert scaling and utility analysis, respectively), appear to capture similar constructs in childhood cancer survivors. PMID- 10679880 TI - Comparison of screening instruments for disability and emotional/behavioral disorders with a generic measure of health-related quality of life in survivors of childhood brain tumors. AB - The sensory, motor, educational and emotional/behavioral outcomes in 32 survivors of childhood brain tumors were evaluated by examination, interview, questionnaires on emotion/behavior and the Health Utilities Index Mark 2 (HUI 2). Thirty-eight percent had moderate/severe disability, and this was associated closely with special educational provision. Pre- and peri-operative factors were the commonest determinants of disability. Fifty percent had a high score on the emotion/behavior questionnaires, suggesting a high risk of an emotional or behavioral problem. The HUI 2 discriminated well between those survivors who had and those who had not had special provision made for their education but poorly between those with high and those with low scores on the emotion/behavior questionnaires. Previous studies have found self-reported health-related quality of life to be related more closely to emotional/behavioral sequelae than to disability. Possible uses and limitations of the HUI 2 in this clinical context are discussed. PMID- 10679881 TI - Cross-cultural adaptation of a health status classification system in children with cancer. First results of the French adaptation of the Health Utilities Index Marks 2 and 3. AB - Our objective was to adapt and validate the Health Utilities Index Mark 2 (HUI 2) and HUI 3 health status classification systems self-report questionnaire in a population of children with cancer, a group of 42 children already included in a multi-centre database designed by the Group on Brain Tumors in Children of the French Society for Pediatric Oncology. Children were recruited during a routine consultation. Most of them had completed treatment. The version of the questionnaire for French adults was adapted linguistically for children. Open ended queries by children about the comprehensiveness of the questions and very low non-response rates showed a good acceptability of the questionnaire. The main psychometric properties of the HUI 2 and HUI 3 classification systems were assessed in 3 groups of raters (child, parent, physician): construct validity was tested against the rating of the child's health state on a Likert scale and through comparison with clinical data, and internal consistency was determined through multi-trait analysis. Weighted and unweighted kappa values were used to measure the inter-rater agreement between the child's, parent's and physician's assessment of the child's health state. The convergent validity was satisfactory, with better results when the physician's assessment was used. The most affected attributes were the expected ones (i.e., cognition, pain and emotion). Disagreement was observed between the 3 raters, more often in the same direction: taking the child's assessment as the reference, the parents tended to under estimate the health status while physicians tended to over-estimate it. PMID- 10679882 TI - Preliminary translation and cultural adaptation of Health Utilities Index questionnaires for application in Argentina. AB - Quality-of-life assessment is being used increasingly in clinical research. This is true particularly in the case of survivors of cancer in childhood, where improving survival rates have raised concern regarding the long-term effects of medical cure. Health-status assessment and quality-of-life instruments have been developed for the most part in the English language, thus necessitating their translation and cultural adaptation for use in non-English-speaking countries. Our purpose was to develop a set of Spanish-language questionnaires for application with a population of children with cancer in a tertiary-care center in Buenos Aires, Argentina. The Health Utilities Index (HUI), a conceptual framework for assessing health status, was chosen for this study. Three distinct questionnaires, based on the HUI, were used: a self-completed one for health professionals and teachers (15Q) to report assessments of children and 2 interviewer-administered ones, for child survivors (42Q) to report assessments about their own health status and parents (45Q) to report assessments about their children's health status. The original translations and reviews were accomplished with direct oversight by members of the HUI Group, to ensure conceptual equivalence. The instruments were then tested in Buenos Aires by application to staff of the hematology-oncology service, childhood cancer patients and the parents of childhood cancer patients. Several modifications were made based on these tests. We concluded that the translation and cultural adaptation of these instruments was adequate for use with the groups tested in a pilot survey of survivors of childhood cancer in Argentina. PMID- 10679883 TI - Research triangulation to derive meaning-based quality-of-life theory: adolescent resilience model and instrument development. AB - We describe the triangulation of qualitative and quantitative research methods used to develop and test the Adolescent Resilience Model (ARM). The differences in meaning-based and function-based health-related quality of life (HRQL) are discussed, and method triangulation is presented as a means of developing models of HRQL that represent the perspectives of the adolescent and family. Qualitative methods of phenomenology, simultaneous concept analysis, focus groups and thematic analysis were used to generate the ARM. Quantitative instrumentation and structural equation model development and testing were used to evaluate the ARM. A decision-making process for combining qualitative and quantitative research, so that both approaches are equally valued and used, is also presented. Int. J. Cancer Suppl. 12:125-131, 1999. PMID- 10679884 TI - Interpreting the meaningfulness of changes in health-related quality of life scores: lessons from studies in adults. AB - The measurement of health-related quality of life (HRQL) in adults with cancer has proceeded more quickly than has similar measurement in children, so there may be value in applying some methods used in adults to studies in children. An example is a health-transition state instrument called the Subjective Significance Questionnaire (SSQ). The SSQ asks patients to give their own estimates of the degree to which their HRQL has changed with time and, thus, provides a method for interpreting the meaningfulness of changes in scores as derived from a general questionnaire, the EORTC QLQ-C30. The development of similar health-transition instruments for children poses special challenges and requires the development of appropriate methodology. It is suggested that, with the use of cartoons and the answers of proxies, it should be feasible to assess the meaningfulness of changes in HRQL over time in young children as well as in adolescents. Int. J. Cancer Suppl. 12:132-137, 1999. PMID- 10679885 TI - Feasibility of implementing health promotion interventions to improve health related quality of life. AB - Survivors of childhood cancer are a growing and vulnerable population. Cure rates for pediatric cancers now exceed 60% and, by the year 2000, an estimated 1 of every 1,000 young adults will be a cancer survivor. Because this population is at increased risk for late medical and neoplastic complications that impact adversely on health-related quality of life, it is important to investigate methods to promote risk reduction by motivating survivors to practice health promoting behaviors. With this background, we initiated a prospective, randomized, controlled feasibility study in which survivors attending a long-term follow-up clinic were randomized to receive standard care or standard care plus an educational intervention. Our objectives were to determine if the intervention would improve the survivors' knowledge about their cancer treatment and risks of late effects and increase their practice of health-protective behaviors. Since July 1995, 272 of 318 families (86%) approached about the study agreed to participate. Of these, 266 are evaluable for assessment of baseline knowledge and health behaviors. Demographic features, baseline knowledge, health perceptions and health behaviors did not differ among randomized groups. Assessment of the intervention's efficacy at changing health behaviors of survivors randomized to the intervention group will be available when the 1-year follow-up evaluations are completed for the study cohort. Our preliminary experience with this pilot study supports the feasibility of educational intervention research in a specialty clinic dedicated to monitoring long-term childhood cancer survivors. Int. J. Cancer Suppl. 12:138-142, 1999. PMID- 10679886 TI - Measuring health-related quality of life in childhood cancer: lessons from the workshop (discussion). PMID- 10679887 TI - Measuring health-related quality of life in pediatric oncology patients: a brief commentary on the state of the art of measurement and application (discussion). PMID- 10679888 TI - Obstacles and opportunities for the use of health-related quality-of-life assessment in pediatric cancer clinical trials (discussion). PMID- 10679889 TI - A postscript to the international workshop on assessing health-related quality of life in children with cancer. PMID- 10679890 TI - Are there two distinct research strategies for developing biologically active molecules: rational design and empirical selection? PMID- 10679891 TI - Native-like cyclic peptide models of a viral antigenic site: finding a balance between rigidity and flexibility. AB - Antigenic site A of foot-and-mouth disease virus (serotype C) has been reproduced by means of cyclic versions of peptide A15, YTASARGDLAHLTTT, corresponding to residues 136-150 of envelope protein VP1. A structural basis for the design of the cyclic peptides is provided by crystallographic data from complexes between the Fab fragments of anti-site A monoclonal antibodies and A15, in which the bound peptide is folded into a quasi-cyclic pattern. Head-to-tail cyclizations of A15 do not provide peptides of superior antigenicity. Internal disulfide cyclization, however, leads to analogs which are recognized as one to two orders of magnitude better than linear A15 in both ELISA and biosensor experiments. CD and NMR studies show that the best antigen, CTASARGDLAHLTT-Ahx-C (disulfide), is very insensitive to environment-induced conformational change, suggesting that cyclization helps to stabilize a bioactive-like structure. PMID- 10679892 TI - Membrane-perturbing activity of Viperidae myotoxins: an electrostatic surface potential approach to a puzzling problem. AB - Phospholipase-like myotoxins are a class of proteins present in Viperidae venom. Despite the high level of amino acid and structural homology with soluble phospholipases A(2), myotoxins are devoid of enzymatic activity and share cytolytic activity by means of a totally unknown mechanism involving the lipid bilayer perturbation. The distribution of electrostatic surface potentials of four myotoxins and seven phospholipases A(2) has been compared. The charge distribution is similar in all active non-cytolytic phospholipases with a strongly positive side corresponding to the domain interacting with the micellar substrate and with the opposite side negatively charged. In contrast, all myotoxins examined are positively charged on both sides. Myotoxin III, the only known example of a myotoxin sharing enzymatic activity, displays the same electrostatic surface potential as other related toxins. Using liposomes made with non-hydrolysable phospholipids, we demonstrate that myotoxin III perturbs the lipid bilayer like other myotoxins. Based on these results, a molecular model for myotoxin-membrane perturbing activity is proposed. In this model, potential double-face binding of myotoxic phospholipases A(2) to lipid surfaces could trigger a lipid bilayer destabilization and could generate a stable fusion pore, probably because of the presence of hydrophobic moieties that flank the cationic sites. PMID- 10679893 TI - Application of surface plasmon resonance for analysis of protein-protein interactions in the G protein-mediated signal transduction pathway. AB - Hundreds of extracellular stimuli are received by cells via the pathways consisting of three basic components: cell-surface receptors, heterotrimeric G proteins, and intracellular effector enzymes or ion channels. A number of additional molecules, including G protein-coupled receptor kinases (GRKs), phosducin and Ca(2+)-binding proteins modulate signal transduction through these cascades. Understanding how these universal pathways work requires a detailed analysis of the interactions between these proteins. The recently emerged technology of surface plasmon resonance (SPR) can study protein-protein interactions by measuring not only the equilibrium binding constants, but also the association and dissociation rates. This article reviews experimental design used by researchers to analyze different components of the G protein pathway by SPR and focuses on the insights this technique provides regarding the kinetics, structure-function aspects and regulation of specific molecular events in the cascade. PMID- 10679894 TI - Conformations of nicotinamide adenine dinucleotide (NAD(+)) in various environments. AB - Enzymes bind NAD(+) in extended conformations and yet NAD(+) exists in aqueous solution as a compact, folded molecule. Thus, NAD(+) conformation is environment dependent. In an attempt to investigate the effects of environmental changes on the conformation of NAD(+), a series of molecular dynamics simulations in different solvents was performed. The solvents investigated (water, DMSO, methanol and chloroform) represented changes in relative permittivity and hydrophobic character. The simulations predicted folded conformations of NAD(+) to be more stable in water, DMSO and methanol. In contrast, extended conformations of NAD(+) were observed to be more stable in chloroform. Furthermore, the extended conformations observed in chloroform were similar to conformations of NAD(+) bound to enzymes. In particular, a large separation between the aromatic rings and a strong interaction between the pyrophosphate and nicotinamide groups were observed. The implications of these observations for the recognition of NAD(+) by enzymes is discussed. It is argued that a hydrophobic environment is important for stabilizing unfolded conformations of NAD(+). PMID- 10679895 TI - Crystal structure of asparagine 233-replaced cyclodextrin glucanotransferase from alkalophilic Bacillus sp. 1011 determined at 1.9 A resolution. AB - The crystal structure of asparagine 233-replaced cyclodextrin glucanotransferase from alkalophilic Bacillus sp. 1011 was determined at 1.9 A resolution. While the wild-type CGTase from the same bacterium produces a mixture of mainly alpha-, beta- and gamma-cyclodextrins, catalyzing the conversion of starch into cyclic or linear alpha-1,4-linked glucopyranosyl chains, site-directed mutation of histidine-233 to asparagine changed the nature of the enzyme such that it no longer produced alpha-cyclodextrin. This is a promising step towards an industrial requirement, i.e. unification of the products from the enzyme. Two independent molecules were found in an asymmetric unit, related by pseudo two fold symmetry. The backbone structure of the mutant enzyme was very similar to that of the wild-type CGTase except that the position of the side chain of residue 233 was such that it is not likely to participate in the catalytic function. The active site cleft was filled with several water molecules, forming a hydrogen bond network with various polar side chains of the enzyme, but not with asparagine-233. The differences in hydrogen bonds in the neighborhood of asparagine-233, maintaining the architecture of the active site cleft, seem to be responsible for the change in molecular recognition of both substrate and product of the mutant CGTase. PMID- 10679896 TI - The conformations of locked nucleic acids (LNA). AB - We have used 2D NMR spectroscopy to study the sugar conformations of oligonucleotides containing a conformationally restricted nucleotide (LNA) with a 2'-O, 4'-C-methylene bridge. We have investigated a modified 9-mer single stranded oligonucleotide as well as three 9- and 10-mer modified oligonucleotides hybridized to unmodified DNA. The single-stranded LNA contained three modifications whereas the duplexes contained one, three and four modifications, respectively. The LNA:DNA duplexes have normal Watson-Crick base-pairing with all the nucleotides in anti-conformation. By use of selective DQF-COSY spectra we determined the ratio between the N-type (C3'-endo) and S-type (C2'-endo) sugar conformations of the nucleotides. In contrast to the corresponding single stranded DNA (ssDNA), we found that the sugar conformations of the single stranded LNA oligonucleotide (ssLNA) cannot be described by a major S-type conformer of all the nucleotides. The nucleotides flanking an LNA nucleotide have sugar conformations with a significant population of the N-type conformer. Similarly, the sugar conformations of the nucleotides in the LNA:DNA duplexes flanking a modification were also shown to have significant contributions from the N-type conformation. In all cases, the sugar conformations of the nucleotides in the complementary DNA strand in the duplex remain in the S-type conformation. We found that the locked conformation of the LNA nucleotides both in ssLNA and in the duplexes organize the phosphate backbone in such a way as to introduce higher population of the N-type conformation. These conformational changes are associated with an improved stacking of the nucleobases. Based on the results reported herein, we propose that the exceptional stability of the LNA modified duplexes is caused by a quenching of concerted local backbone motions (preorganization) by the LNA nucleotides in ssLNA so as to decrease the entropy loss on duplex formation combined with a more efficient stacking of the nucleobases. PMID- 10679897 TI - FDG PET evaluation of head and neck cancer: value of imaging the thorax. AB - BACKGROUND: Patients with primary tumors of the head and neck have been reported to have a high rate of synchronous primary tumors of the upper aerodigestive tract. This study was performed to determine whether inclusion of the thorax in the scan volume would be diagnostically useful for positron emission tomography (PET) with [F-18] fluorodeoxy-D-glucose (FDG) in patients with primary tumors of the head and neck. METHODS: FDG PET scans from the midcranium to the diaphragm were obtained on 56 patients with a variety of head and neck tumors on initial examination before definitive therapy. PET findings in the chest were correlated with results of all other imaging studies, biopsy results, and clinical follow up. RESULTS: In nine studies (16%), areas of increased FDG uptake in the chest were seen and were judged to be tumors. Six of these probably were false-positive results, although one of these six may have been unconfirmed true positives. Of the three confirmed true-positive studies, two were obvious from other routine studies. In only one case did the PET study reveal a significant lesion not found by means of routine evaluation, resulting in a case-finding yield of 2%. If the unconfirmed possible true-positive results are included, the case-finding yield increases to 4%. CONCLUSIONS: No compelling indication was seen for including the chest in PET studies of patients with head and neck cancer. PMID- 10679898 TI - The fate of osseointegrated implants in patients following oral cancer surgery and mandibular reconstruction. AB - BACKGROUND: The feasibility of implant treatment in patients after oral ablative tumor surgery and defect reconstruction has not yet been investigated in terms of the requisite high standards of success assessment. A report on this topic must address not only implant survival but implant health, bone response, soft tissue health, failure pattern, and time of failure, as well. METHODS: From June 1990 through December 1997, 90 patients received 320 dental implants after oral tumor resection and immediate soft tissue reconstruction. Included in the study were 45 patients with 162 implants loaded for at least 1 year. Regular follow-up for 6 years consisted of detailed medical history and evaluation of periodontal parameters. Out of this population, 10 vascularized iliac bone grafts for mandibular reconstruction containing loaded implants were selectively evaluated for bone loss. RESULTS: The assessment of pocket probing depths, plaque accumulation, bleeding disposition, implant mobility by means of the Periotest method applied to the restoration type, horizontal and vertical (peri-implant) bone loss according to x-ray findings, causes and time of implant loss, and subjective statements offered results comparable to those found in healthy subjects examined with periodontal success parameters. CONCLUSION: Prosthetic restoration of patients after oral ablative tumor surgery followed by hard and soft tissue reconstruction can be achieved with dental implants with similar long term efficacy as found in healthy subjects adhering to internationally established requirements. PMID- 10679899 TI - Impact of tongue base and posterior pharyngeal wall biomechanics on pharyngeal clearance in irradiated postsurgical oral and oropharyngeal cancer patients. AB - BACKGROUND: Postsurgical oral and oropharyngeal cancer patients may experience pharyngeal clearance problems after completion of postoperative radiotherapy. METHODS: Swallowing was examined in six patients using videofluoroscopy for up to 1 year after surgery. Biomechanical analysis was used to mark movement of the tongue base and posterior pharyngeal wall during swallowing. RESULTS: The majority of patients experienced increased problems with pharyngeal clearance at or after their 6 month posthealing evaluation, generally 18 to 22 weeks after completion of radiotherapy. Pharyngeal residue was associated with a disruption in either tongue base or posterior pharyngeal wall movement. CONCLUSIONS: Increased fibrosis of the pharyngeal musculature after completion of radiotherapy may have a negative impact on pharyngeal clearance in addition to any pharyngeal clearance problems resulting from surgical resection. Tongue base to posterior pharyngeal wall contact is essential but not sufficient for effective pharyngeal clearance. Sufficient duration of tongue base to posterior pharyngeal wall contact is also needed to provide adequate pharyngeal bolus driving pressure. PMID- 10679900 TI - Reliability and validity of an observer-rated disfigurement scale for head and neck cancer patients. AB - BACKGROUND: Facial disfigurement is considered to be one of the most distressing aspects of head and neck cancer and its treatment, but it has been the focus of little systematic study. Existing studies have yielded conflicting results about the psychosocial impact of disfigurement. No studies to date have examined disfigurement using a valid and reliable observer-rated measure. The purpose of the current study was to examine the validity (convergent and discriminant) and the inter-rater reliability of a novel nine-point observer-rated disfigurement scale. METHODS: The sample consisted of 74 ambulatory head and neck cancer patients more than 6 months post treatment. Ratings of disfigurement were assigned independently by surgical and nonsurgical raters. Validity was assessed by comparing the association between disfigurement ratings and sociodemographic and illness treatment variables. Reliability was assessed by examining the concordance between the surgical and nonsurgical ratings. RESULTS: Disfigurement ratings were not associated with several sociodemographic variables, supporting the discriminant validity of the scale. Disfigurement was significantly related to a diagnosis of oral cancer, a history of adjunctive radiation, the type of surgical procedure performed, the degree of physical dysfunction, and the presence of postoperative complications. Observer ratings of disfigurement were significantly related to patient ratings of disfigurement. These findings support the convergent validity of the disfigurement scale. Inter-rater reliability of the scale was high (intraclass correlation coefficient =.91). CONCLUSION: The study provides preliminary evidence for the validity and inter-rater reliability of a novel nine point observer-rated disfigurement scale that may be useful in evaluating the impact of disfigurement on quality of life in head and neck cancer. PMID- 10679901 TI - Experience with Barton button and peristomal breathing valve attachments for hands-free tracheoesophageal speech. AB - BACKGROUND: Tracheostoma breathing valves permit hands-free tracheoesophageal (TE) speech production; however, few laryngectomees routinely use them because of problems with attachment. METHODS: We retrospectively reviewed the charts of 45 TE speakers to determine the success rate and factors associated with successful breathing valve use based on attachment. All patients attempted to use a tracheostoma breathing valve with either a standard or customized peristomal housing, or a standard or customized Barton button. Device selection was based on inspection of the patient's neck and peristomal contour. Six to eight consecutive hours of attachment defined success. RESULTS: Overall, 9% of subjects succeeded with any peristomal attachment as compared to 68% with either a standard (57%) or customized (85%) Barton button. Smooth stomal contour, a contiguous stomal lip, and correct button length were important for successful Barton button use. CONCLUSIONS: Standard or customized Barton buttons offer excellent alternatives to peristomal housing attachments for hands-free TE speech in select patients. PMID- 10679902 TI - Effects of arachidonic acid metabolites in a murine model of squamous cell carcinoma. AB - BACKGROUND: A murine model (C3H mice) of squamous cell carcinoma (SCCVII) has been used to investigate the role of arachidonic acid (AA) metabolites in head and neck cancer. Inhibition of tumor growth by cyclooxygenase (COX) and lipoxygenase (LOX) inhibitors of AA metabolism has been associated with changes in levels of AA metabolites in tumor tissues and inflammatory cell infiltrates. To characterize this model further, the effects of exogenous AA metabolites on tumor growth in vitro and in vivo were investigated. METHODS: Following subcutaneous inoculation with SCCVII tumor cells, control (16 mice) and treatment (24 mice) groups were injected with peritumoral vehicle or AA metabolite. Peritumoral injections of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and 12 hydroxyeicosatetraenoic acid (12-HETE) were performed for 16-21 days, and final excised tumor weights were measured. In vitro production of PGE2 and LTB4 was assayed in 2-5 day cultures of SCCVII. Exogenous PGE2 effects on tumor cell growth was assessed with the MTT assay in vitro. RESULTS: Tumor growth was significantly inhibited (p =.03) following peritumoral injection of PGE2. Final tumor weights were not affected by LTB4 or 12-HETE. Tumor inhibition by PGE2 was associated with increased tumor tissue levels of LTB4 (p =.04). In vitro, SCCVII produced minimal amounts of PGE2 and LTB4, and PGE2 had minimal effect on growth. CONCLUSIONS: In this model, tumor inhibition by exogenous PGE2 is primarily mediated by affecting host-tumor interactions, although there may be some direct effect on tumor cells. Changes in tumor tissue levels of LTB4 following peritumoral PGE2 administration may be attributable to negative feedback inhibition of the COX pathway with shunting into the LOX pathway. SCCVII cells are probably not a significant source of prostaglandins and leukotrienes in vivo. These data provide insight into the mechanism of action of inhibitors of AA metabolism on tumor growth. PMID- 10679903 TI - Multicentricity in pharyngoesophageal tumors: argument for total pharyngolaryngoesophagectomy and gastric transposition. AB - BACKGROUND: Pharyngoesophageal tumors pose a challenge to surgical management, and there is controversy in the literature as to the best procedure to be used. Advantages and disadvantages are mentioned for total pharyngolaryngoesophagectomy and gastric transposition (PLE>), free jejunal transplants, and free forearm flaps. One of the arguments for PLE> is the persistence or subsequent occurrence of multiple primaries in a field cancerization region. Multiple tumors in the head and neck/esophagus/lung axis have been reported. However, despite extensive investigation, there is little information on specific multicentricity in patients treated with PLE> for pharyngolaryngoesophageal carcinomas. METHODS: A clinicopathological study was undertaken in 35 consecutive patients who underwent PLE> for pharyngoesophageal cancer to evaluate synchronicity, multicentricity, and metachronicity. Only in situ and invasive carcinomas were considered. The findings were compared with the reports in the literature. RESULTS: Thirty-eight tumors were diagnosed preoperatively, with the main indications for PLE> being tumors located in the esophagus or hypopharynx (32 patients) and larynx (three patients). After the surgical treatment, 21 patients had single primaries (60%) and 14 (40%) had 25 multiple primaries in addition to their main primaries (total of 60 tumors in the whole group). Synchronous, previous metachronous and subsequent metachronous carcinomas occurred in 26%, 17%, and 8.5% of the instances, respectively. Twenty of the 25 multicentric carcinomas were invasive (80%). Either the main primaries or the multicentric carcinomas were located in the esophagus or hypopharynx (91.5% and 60%, respectively). Other sites included the larynx, oropharynx, oral cavity, and lung. CONCLUSION: The incidence of multicentric tumors in patients with pharyngoesophageal carcinomas may favor total PLE> as the procedure of choice, because it includes all the condemned upper pharyngolaryngoesophageal mucosa. PMID- 10679904 TI - Squamous cell carcinoma of maxillary sinus. AB - BACKGROUND: Medical records of 43 patients with histologically proved diagnosis of squamous cell carcinoma who were treated between the years 1975 and 1994 at the department of Otolaryngology Head Neck Surgery, VU Amsterdam were examined. METHODS: Tumors were restaged according to UICC classification 1997. Thirty-eight patients were treated for cure, nine were treated with chemotherapy followed by external beam radiotherapy, and 28 patients were treated with surgery followed by postoperative radiotherapy. No patient was lost to follow-up. Data with respect to survival were analyzed. RESULTS: Eighty-three percent of the tumours were in stage III or stage IV at the time of first presentation. Five-year survival after surgery and postoperative radiotherapy for all patients was 64%. For stages II, III, and IV it was 83%, 49%, and 37%, respectively. Cervical nodal metastases were present in 4.1% at the time of presentation. Thirty-seven percent of the patients survived 2 years after chemotherapy followed by radiotherapy. CONCLUSIONS: Squamous cell carcinoma continues to be diagnosed late. Surgery followed by radiotherapy remains the treatment of choice. Mandibulotomy should be considered for better clearance of retromaxillary space in T3 -T4 tumors. The eye should be preserved whenever it is oncologically safe to do so. PMID- 10679905 TI - Diagnosis of superficial esophageal cancer and dysplasia using endoscopic screening with a 2% lugol dye solution in patients with head and neck cancer. AB - BACKGROUND: Head and neck cancer (HNC) has a high incidence in Brazil, with cancer of the oral cavity being one of the five most common cancers among Brazilians. Alcohol and tobacco consumption may contribute to synchronous or metachronous HNC and esophageal cancer. The early detection of superficial esophageal cancer and dysplasia in asymptomatic patients with HNC, after successfully treating the primary cancer, may provide an effective cure. METHODS: A prospective study involving 60 patients with HNC was carried out at the State University of Campinas (UNICAMP) to screen for superficial esophageal cancer and dysplasia using endoscopy and a 2% lugol dye solution followed by biopsy of the suspicious areas. RESULTS: Five patients (8.3%) had superficial esophageal cancer, which was diagnosed as intraepithelial carcinoma in three of them (5.0%). In four patients, the superficial esophageal cancer was synchronous, and in one it was metachronous to HNC. Five patients (8.3%) had dysplasias in the esophageal epithelium (three were classified as mild and two as moderate). CONCLUSION: These results demonstrate the value of endoscopic screening of the esophagus using lugol dye in patients with HNC, particularly because superficial esophageal cancer is extremely difficult to detect by conventional methods in asymptomatic patients. PMID- 10679906 TI - The performance of SF-36 health survey in patients with laryngeal cancer. Head and Neck Cancer Italian Working Group. AB - BACKGROUND: Interest in measuring health-related quality of life has increased together with the awareness that such humanistic outcomes require valid and reliable measures. Among the several generic questionnaires, the Short Form 36 Items Health Survey (SF-36) is recognized for its comprehensiveness, brevity, and high standards of reliability and validity. It has been translated and validated in several languages. METHODS: In the framework of a larger, prospective, multicenter study aimed to produce and validate an Italian questionnaire tailored to laryngeal cancer patients, the SF-36 was administered to a sample of well characterized cases. It was, therefore, possible to test its characteristics in terms of patients' acceptance, psychometric, and clinical validity. RESULTS: Overall, findings show that in this sample of 165 consecutive patients with laryngeal cancer at various stage of disease, the SF-36 performance was very good. The patients' acceptance was satisfactory: all patients completed the questionnaire. All the questionnaire scales met the standards suggested in terms of grouping and scaling assumptions. The internal reliability coefficients actually replicate the satisfactory findings reported for the original SF-36. In terms of capability of the questionnaire scales to discriminate between groups expected to differ in a given health concept in relation to clinical variables, the results were also good. CONCLUSIONS: This study showed that SF-36 was well accepted by patients and was able to detect the impact of different treatment approaches on health- related quality of life. It is likely that the sensitivity and the precision of the SF-36 can be further improved by integrating brief questionnaire modules specific for laryngeal clinical issues. PMID- 10679907 TI - Primary melanocytic schwannoma of cervical sympathetic chain. AB - BACKGROUND: Primary melanocytic schwannoma arising from the cervical sympathetic chain is a rare pigmented nerve sheath tumor. Two cases are presented from an academic medical center. Patients and Methods Patients were initially seen with an enlarging neck mass associated with sympathetic nervous system dysfunction. Radiography demonstrated a mass located posterior to the carotid sheath. Primary therapy consisted of surgical excision and postoperative radiation therapy. RESULTS: The tumors were found to be melanocytic schwannomas arising from the cervical sympathetic chain. The pathologic characteristics of this neoplasm are reviewed. One patient remained disease free for 12 years after treatment, whereas 1 patient died as a result of local recurrence and distant metastases. CONCLUSIONS: Melanocytic schwannoma of the cervical sympathetic chain is a rare nerve sheath tumor of the head and neck that may be misdiagnosed as malignant melanoma. The clinical behavior of this neoplasm is variable. Preoperative neurologic findings, anatomic location, electron microscopy, and immunohistochemistry findings help to establish the diagnosis, and electron microscopy may have a role in distinguishing between benign and malignant lesions. Complete surgical excision is the treatment of choice. PMID- 10679908 TI - Expression of basic fibroblast growth factor protein and its down-regulation by interferons in head and neck cancer. AB - BACKGROUND: Angiogenesis is crucial for tumor growth and metastasis. In several tumors, microvascular density has been shown to correlate with metastasis and aggressiveness. Basic fibroblast growth factor (bFGF) has potent angiogenic activity and has been identified in a wide variety of malignancies including head and neck squamous cell carcinomas (HNSCC). Material and Methods Frozen sections of 50 HNSCC were immunostained for von Willebrand factor and bFGF. Microvessels were counted by light microscopy; bFGF expression was studied at the light and electron microscopic level. Laryngeal cancer cell line HlaC79 was incubated with interferon (IFN) alpha and beta. bFGF quantification was performed by ELISA, and antiproliferative effects were determined by BrdU assay. RESULTS: The mean number of blood vessels (77.5 +/- 23.7) is significantly increased in HNSCC compared with controls (17.1 +/- 5.9). bFGF protein expression was detected in all HNSCC but not in control tissue. An correlation between bFGF expression and mean number of microvessels was found (p <.001). However, no correlation between bFGF expression and the main clinicopathologic features was shown. The long-term exposure (144 hr) of HNSCC cells to noncytostatic concentrations of IFN alpha and beta (>10 U/mL) down-regulated the protein production of bFGF. CONCLUSION: bFGF expression and angiogenesis are enhanced in HNSCC. The higher microvessel density in HNSCC with strong bFGF expression supports the importance of bFGF for tumor angiogenesis. IFN alpha and beta treatment leads to a down-regulation of bFGF expression independent of their antiproliferative effects, suggesting that IFN treatment might result in a reduction of angiogenesis in HNSCC. PMID- 10679909 TI - Carcinoma of the dorsum of the tongue. AB - BACKGROUND: Squamous cell carcinoma (SCC) of the anterior two thirds of the tongue is the second most common oral cancer, with the lateral border being the most common location. Squamous cell carcinoma of the dorsum of the tongue is exceedingly rare and has been described in the past as a myth or misdiagnosis. The clinical diagnosis of SCC on the dorsum of the tongue is difficult because it may be mimicked by a wide variety of benign and premalignant lesions, including granular cell myoblastoma, erosive lichen planus, medial rhomboid glossitis, and amyloidosis. In this study we re-evaluate the entity of SCC of the dorsum of the tongue. METHODS: We reviewed 5 large series of carcinoma of the tongue, which include accurate documentation of the topographic location of the carcinoma. We also describe 5 cases of SCC of the dorsum of the tongue from our own series of 99 patients with carcinoma of the tongue. RESULTS: In all 6 series, carcinoma of the dorsum of the tongue was present in 3 to 5% of the carcinomas of the tongue. CONCLUSION: Although rare, SCC of the dorsum of the tongue exists and may be mimicked by benign conditions, thus SCC should be suspected when diagnosing lesions of this area of the tongue. PMID- 10679910 TI - Identification of novel regions of allelic loss from a genomewide scan of esophageal squamous-cell carcinoma in a high-risk Chinese population. AB - Esophageal cancer is one of the most common fatal cancers worldwide. Deletions of genomic regions are thought to be important in esophageal carcinogenesis. We conducted a genomewide scan for regions of allelic loss using microdissected DNA from 11 esophageal squamous-cell carcinoma patients with a family history of upper gastrointestinal tract cancer from a high-risk region in north central China. Allelic patterns of 366 fluorescently labeled microsatellite markers distributed at 10-cM intervals over the 22 autosomal chromosomes were examined. We identified 14 regions with very high frequency (>/= 75%) loss of heterozygosity (LOH), including broad regions encompassing whole chromosome arms (on 3p, 5q, 9p, 9q, and 13q), regions of intermediate size (on 2q, 4p, 11p, and 15q), and more discrete regions identified by very high frequency LOH for a single marker (on 4q, 6q, 8p, 14q, and 17p). Among these 14 regions were 7 not previously described in esophageal squamous-cell carcinoma as having very high frequency LOH (on 2q, 4p, 4q, 6q, 8p, 14q, and 15q). The very high frequency LOH regions identified here may point to major susceptibility genes, including potential tumor suppressor genes and inherited gene loci, which will assist in understanding the molecular events involved in esophageal carcinogenesis and may help in the development of markers for genetic susceptibility testing and screening for the early detection of this cancer. Genes Chromosomes Cancer 27:217 228, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10679911 TI - Identification of breakpoint cluster regions at 1p36.3 and 3q21 in hematologic malignancies with t(1;3)(p36;q21). AB - The reciprocal translocation t(1;3)(p36;q21) is associated with myelodysplastic syndromes (MDSs) and acute myeloid leukemia (AML) characterized by trilineage dysplasia, in particular dysmegakaryocytopoiesis, and a poor prognosis. As yet no molecular genetic analyses of the t(1;3) have been reported. In four patients with t(1;3), all of whom had AML-M4, which evolved from MDS, the breakpoints at 3q21 clustered within a 60-kb region centromeric to the breakpoint of the inv(3)(q21q26), whereas the breakpoints at 1p36 clustered within a 90-kb region at 1p36.3. The presence of novel clusters in both the 3q21 and 1p36 breakpoints (BCRs) suggests a common, underlying molecular mechanism for the development of t(1;3)-positive MDS/AML. The Ribophorin I (RPN1) gene close to the BCR at 3q21 was highly expressed without gross structural changes, whereas the GR6 gene located within the BCR at 3q21 was not expressed. No other highly expressed genes were isolated in a 150-kb region at 3q21. Thus, it is likely that a gene at 1p36.3 is activated by the translocation of the 3q21 region or a gene important for transformation lies on 3q21, outside the 150-kb region. Further characterization of the BCRs at 1p36.3 and 3q21 should provide important insights into the molecular genetic mechanisms involved in the genesis of t(1;3)-positive MDS/AML. Genes Chromosomes Cancer 27:229-238, 2000. PMID- 10679912 TI - Genomic FHIT analysis in RER+ and RER- adenocarcinomas of the pancreas. AB - Alterations of the candidate tumor suppressor gene FHIT have been reported in multiple tumor types, including pancreatic carcinoma. The mechanism of FHIT genomic inactivation is unusual, most frequently occurring by homozygous deletion, whereas only rare cases have missense mutations. Altered (shortened) transcripts and reduced protein expression are reported, but a genetic basis for these is often inapparent. We studied FHIT genomic alterations of pancreatic carcinomas. Loss of heterozygosity (LOH) was found in 41% of 93 carcinomas without microsatellite instability (RER(-)), but no mutations were found by genomic sequencing. Homozygous deletions inside the FRA3B fragile site were found in four RER(-) tumors, but only two affected the FHIT coding region. In contrast, FHIT alterations were found in the three RER(+) pancreatic carcinomas screened; two had FHIT homozygous deletions affecting exon 5 and the third had a heterozygous missense mutation (H76N). The excess occurrence of homozygous deletions at this site in RER(+) pancreatic cancers is statistically significant (P < 0.01). Since homozygous deletions have not previously been reported in RER(+) carcinomas at any genomic site, an extremely high rate of site-specific deletion must exist within the FRA3B-related FHIT gene. Consequently, the paucity of documented inactivating point mutations cannot be used to judge the presence or absence of putative FHIT-related selective pressures that act during tumorigenesis of RER(-) neoplasia. Nonetheless, the identification of a heterozygous mutation as the sole sequence abnormality might raise doubt as to whether strong selective pressures are afforded by FHIT genomic inactivation in this tumor type. Genes Chromosomes Cancer 27:239-243, 2000. PMID- 10679913 TI - Allelotyping of anaplastic thyroid carcinoma: frequent allelic losses on 1q, 9p, 11, 17, 19p, and 22q. AB - Little is known about the genetic mechanisms behind the genesis of anaplastic thyroid carcinoma. This is among the most virulent of all human malignancies, and it is believed to result most often from transformation of differentiated thyroid carcinomas of the papillary type. So far, TP53 and beta-catenin mutations are the only genetic alterations that have been implicated in its pathogenesis. To identify loci of other potential tumor suppressor genes, we carried out a genome wide allelotyping study using 39 microsatellite markers representing all nonacrocentric autosomal arms, in a panel of 21 anaplastic thyroid carcinomas. Frequent allelic losses were identified in 1q (40%), 9p (58%), 11p (33%), 11q (33%), 17p (44%), 17q (43%), 19p (36%), and 22q (38%). Deletion mapping of chromosome arms with the most frequent allelic losses (frequencies above 40%) localized the commonly deleted region to 1q31-42, 9p21-22, 17p12-ter, and 17q21.1 22. The mean frequency of loss of heterozygosity on all arms tested was 20%, and the mean fractional allelic loss among the cancers examined was 0.20. These findings defined a sharp distinction between anaplastic thyroid carcinomas and papillary thyroid carcinomas, because the latter do not tend to show losses at the same loci. Frequent allelic losses at multiple loci may implicate chromosomal instability as an important factor in the development of anaplastic thyroid carcinomas. Genes Chromosomes Cancer 27:244-251, 2000. PMID- 10679914 TI - Genetic and phenotypic changes associated with the acquisition of tumorigenicity in human bladder cancer. AB - There has been a general lack of human paired cell lines that both reproduce the in vivo spectrum of tumor progression of bladder cancer and have some of the genetic changes associated with progression in human tumor tissue. T24, a cell line established from an invasive human transitional cell carcinoma (TCC) of the bladder, has been used extensively in bladder cancer research. However, a significant limitation of this cell line is its lack of tumorigenicity when injected into immunocompromised mice. This characteristic was used to our advantage as we sought to characterize T24T, a highly tumorigenic variant that could then be used to elucidate the genes responsible for human bladder tumor progression. In culture, T24T has a faster doubling time, reaches a higher cell density in monolayer culture, and is more motile than T24 at higher cell densities. T24T is able to form colonies in soft agar, whereas T24 is not, and expresses HRAS, a gene associated with increased aggressiveness in human TCC, at higher levels than T24. Most importantly, T24T forms solid tumors when injected subcutaneously in SCID mice both with and without Matrigel (Sigma, St. Louis, MO), whereas T24 does not. Cytogenetically, the 2 cell lines contain at least 5 shared structural anomalies, as determined by detailed karyotyping. Interestingly, T24T has acquired 4 new structural changes, 3 of which [add(10)(p12), i(10)(q10), -15] have been observed in loss of heterozygosity (LOH) studies of tumor progression in human TCC. It appears that the T24/T24T model may be an excellent tool for the study of human TCC progression because of its relationship with known karyotypic changes associated with human bladder cancer progression. We are currently taking advantage of these paired cell lines to identify genes involved in human TCC progression. Genes Chromosomes Cancer 27:252-263, 2000. PMID- 10679915 TI - MLL-CBP fusion transcript in a therapy-related acute myeloid leukemia with the t(11;16)(q23;p13) which developed in an acute lymphoblastic leukemia patient with Fanconi anemia. AB - We describe a boy with Fanconi anemia (FA) who developed acute lymphoblastic leukemia (ALL) (FAB-LI) followed by acute myeloid leukemia (AML) (FAB-M5) at relapse. The patient was diagnosed with early pre-B-cell ALL without preceding aplastic anemia and was treated with ALL-oriented chemotherapy which included doxorubicin (a total dose of 140 mg/m(2) administered), which is a topoisomerase II inhibitor. Complete remission was obtained, but after 38 weeks AML developed. The karyotype of ALL cells at diagnosis showed 46,XY, and that of AML cells at relapse was 46,XY, t(11;16)(q23;p13). An MLL gene rearrangement and MLL-CBP chimeric mRNA were found in AML, but not in ALL. A diagnosis of FA was confirmed by an increased number of chromosomal breaks and rearrangements in peripheral blood lymphocytes cultured with mitogen in the presence of mitomycin C. We conclude that this FA patient developed ALL followed by a therapy-related t(11;16)-AML resulting in an MLL-CBP fusion. Further examination of such patients would shed light on leukemogenesis in FA patients. Genes Chromosomes Cancer 27:264-269, 2000. PMID- 10679916 TI - Modification in the inherent mode of allelic replication in lymphocytes of patients suffering from renal cell carcinoma: a novel genetic alteration associated with malignancy. AB - Using fluorescence in situ hybridization (FISH) to interphase nuclei, we examined the replication timing of 1 allele relative to its counterpart in PHA-stimulated peripheral blood lymphocytes of normal subjects and patients suffering from a solid tumor (renal cell carcinoma). In the FISH assay, an unreplicated DNA sequence is identified by a single dot-like hybridization signal, whereas a replicated region gives rise to a duplicated, bipartite signal. Accordingly, lymphocytes of normal individuals show 2 patterns of allelic replication: (i) synchronized replication of allelic counterparts, as exemplified by the biallelically expressed loci TP53 and D21S55; and (ii) non-synchronized replication of allelic partners, as exemplified by the early and late replicating alleles of GABRB3, an imprinted locus subjected to monoallelic expression. However, when present in lymphocytes of the cancer patients, all 3 loci change their replication mode: alleles of TP53 and D21S55 become asynchronous, whereas the early replicating allele of GABRB3 delays replication, leading to relaxation in the imprinted mode of replication. Based on the tight relationship between temporal order of allelic replication and allelic mode of expression, the modified order of allelic replication observed in nonmalignant cells of individuals diagnosed with cancer represents a novel genetic alteration associated with malignancy. This alteration detected by simple cytogenetic means, applied to peripheral blood lymphocytes, offers a potential test for cancer identification. Genes Chromosomes Cancer 27:270-277, 2000. PMID- 10679917 TI - Non-random involvement of chromosome 13 in patients with persistent or relapsed disease after bone-marrow transplantation for chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) patients with persistent or relapsed disease following bone-marrow transplantation (BMT) usually show both clonal and non clonal cytogenetic changes in addition to the Philadelphia (Ph) translocation. These changes are presumably due to conditioning prior to transplantation and are generally not thought to be of clinical significance. We have examined the additional cytogenetic changes found in Ph+ve cells after BMT in 47 CML patients. Forty patients showed clonal changes. The involvement of each chromosome was compared statistically with expected values assuming that further chromosome changes are random and related to chromosome size. In clones that comprised 50% or more of the Ph+ve metaphases, chromosome 13 was involved in 12 of 22 clones (55%); this was highly significant when compared with the theoretical expected value of 3.2 (14.5%) (P < 0.001). The chromosome 13 rearrangements comprised both translocations and deletions. By means of FISH with a panel of 13q YAC clones, the breakpoints in 6 of these patients were investigated, but no common site of translocation was identified. The YAC panel was then used on material from 6 patients with chromosomal deletions. A common region of deletion was identified at 13q12-14, suggesting the presence of one or more tumor suppressor genes. We conclude that chromosome 13 deletions are non-randomly overrepresented in Ph+ve metaphases following BMT for CML. Genes Chromosomes Cancer 27:278-284, 2000. PMID- 10679918 TI - t(11;14)-positive mantle cell lymphomas exhibit complex karyotypes and share similarities with B-cell chronic lymphocytic leukemia. AB - Until now, few data on additional chromosomal aberrations in t(11;14)-positive mantle cell lymphomas (MCLs) have been published. We analyzed 39 t(11;14) positive MCLs by either comparative genomic hybridization (CGH; n = 8), fluorescence in situ hybridization (FISH) with a set of DNA probes detecting the most frequent aberrations in B-cell neoplasms (n = 12), or both techniques (n = 19). The t(11;14) was present in all cases. In 37 of 39 cases, chromosomal imbalances were found. In 27 cases, complex karyotypes, i.e., >/= 3 aberrations, were identified. The most frequent aberrations were losses of 13q14-21 or 13q32 34 (27 cases), 9p21 (16 cases), and 11q22-23 (12 cases) and gains of 3q26-29 (19 cases), 8q22-24 (11 cases), and 18q21-22 (9 cases). In 26% of cases (7 of 27) analyzed by CGH, a total of 10 high-level DNA amplifications were identified. Although in comparison with B-cell chronic lymphopcytic leukemia (B-CLL) MCL is characterized by a much higher complexity of chromosomal aberrations, there are striking similarities: 13q14 deletions were identified in more than 50% of both MCL and B-CLL cases. In contrast, in our CGH database containing 293 B-cell lymphomas, this aberration was found in only 11% of other nodal lymphomas. Even more strikingly, 11q deletions, which are present in 20%-30 % of MCL and B-CLL, were found very rarely in other nodal B-cell lymphomas (CGH: 1 of 208 cases; FISH: 1 of 69 cases). These data show that MCL is characterized by specific secondary aberrations and that there may be similarities in the pathogenesis of MCL and B-CLL. Genes Chromosomes Cancer 27:285-294, 2000. PMID- 10679919 TI - A sporadic breast tumor with a somatically acquired complex genomic rearrangement in BRCA1. AB - Germ-line mutations in BRCA1 cause a substantial proportion of inherited breast cancer, and most result in inactivated BRCA1 proteins upon translation. Tumours developing in BRCA1 mutation carriers generally show loss of the wild-type allele. However, acquired inactivating mutations in BRCA1 in non-inherited breast tumours showing loss of heterozygosity at the gene locus have not been detected so far. Here we provide evidence that such mutations can be detected in a small proportion of breast tumours. Prompted by recent reports of Alu-mediated large genomic rearrangements in BRCA1, we have investigated whether such rearrangements might occur in sporadic breast cancer as well and have been missed thus far by traditional PCR-based mutation screening technology. To this end, we performed Southern blot analysis of 81 apparently sporadic breast tumours using probes covering exons 6-24 and 3 restriction enzymes. We identified 1 case with an acquired rearrangement (1.2%), indicating that BRCA1 inactivation through changes in the primary genomic sequence of the gene is uncommon in breast cancer. Genes Chromosomes Cancer 27:295-302, 2000. PMID- 10679920 TI - Novel gene fusion of COX6C at 8q22-23 to HMGIC at 12q15 in a uterine leiomyoma. AB - Cytogenetic analyses have shown that aberrations involving 12q13-15 are frequent chromosomal changes in a variety of human benign mesenchymal tumors, e.g., pleomorphic adenomas of the parotid gland, pulmonary chondroid hamartomas, lipomas, and uterine leiomyomas. Recently, the high-mobility group protein gene HMGIC was identified as the target gene affected by the 12q13-15 aberrations. Using 3' rapid amplification of cDNA ends experiments, we isolated novel ectopic sequences fused to HMGIC in a uterine leiomyoma. Cloning of the fusion cDNA identified the human cytochrome c oxidase subunit VIc (COX6C) gene on 8q22-23 as the fusion partner of HMGIC. Nucleotide sequences of the fusion transcript revealed that the first 3 exons of the HMGIC gene, encoding the 3 DNA binding domains, was fused to the exon 2 of the COX6C gene. The identification of a gene rearrangement suggests a role for HMGIC in tumorigenesis of uterine leiomyoma and suggests a possible involvement of HMGIC in mesenchymal differentiation. Genes Chromosomes Cancer 27:303-307, 2000. PMID- 10679921 TI - Two regions of homozygous deletion clusters at chromosome band 9p21 in human lung cancer. AB - We examined 149 lung cancer cell lines for homozygous deletions using 24 DNA markers, which were mapped and ordered in chromosome band 9p21, to define the target regions for 9p21 deletions in human lung cancer. Homozygous deletions were detected in 39 (26%) cell lines and clustered at 2 independent regions. One was the region containing the p16/CDKN2A tumor suppressor gene, and this region was deleted in 32 (21%) cell lines. The other was the region containing D9S171, which is the locus approximately 3 Mb proximal to the CDKN2A locus. This region, designated as the D9S171 region, was deleted in 18 (12%) cell lines. Seven of the 18 cell lines had identical minimum deletions of a 17,036 bp sequence located 20 kb distal to the D9S171 locus. However, such a deletion was also observed in the corresponding B-lymphoblastoid cell line from 1 of the 7 cell lines and in 5 (16%) of 32 noncancerous tissues, suggesting that the deletion was a genetic polymorphism. By considering this polymorphism, 11 (7%) cell lines still had deletions at the D9S171 region. Two NSCLC cell lines showed deletions at the D9S171 region and retentions of the CDKN2A locus. Furthermore, an NSCLC cell line showed discontinuous deletions including either the CDKN2A or D9S171 locus. Therefore, the region surrounding the D9S171 locus was defined as another target region for the 9p21 deletions. It is possible that unknown tumor suppressor gene(s) are present in this chromosomal region. Genes Chromosomes Cancer 27:308 318, 2000. PMID- 10679922 TI - Molecular analysis of the candidate tumor suppressor gene ING1 in human head and neck tumors with 13q deletions. AB - The candidate tumor-suppressor gene ING1 encodes p33(ING1), a nuclear protein which physically interacts with TP53. It has been shown that p33(ING1) acts in the same biochemical pathway as TP53, leading to cell growth inhibition. Interestingly, a rearrangement of the ING1 gene was found in a neuroblastoma cell line, supporting its involvement in tumor development. Because ING1 resides on the long arm of chromosome 13 (13q34) (a region frequently deleted in many tumor types), we sought to characterize its role in head and neck squamous-cell carcinoma (HNSCC). We first analyzed 44 primary tumors for loss of heterozygosity (LOH) at 13q, using four widely spaced microsatellite markers (13q14, 13q14.3 q22, 13q22, and 13q34). Twenty (48%) of the tumor samples showed LOH in all of the informative markers tested, including D13S1315 at 13q34. Two of the tumors displayed partial losses restricted to one marker (D13S118 at 13q14 in tumor 1164, and D13S135 at 13q14.3-q22 in tumor 1398). We then determined the genomic structure of the ING1 gene and sequenced the entire coding region in 20 primary tumors showing 13q LOH and in five head and neck cancer cell lines. A single germline polymorphism was detected in 10 of the tumors analyzed (T to C change) located 110 nucleotides upstream of the starting methionine. No somatic mutations were found in any of the samples, suggesting that ING1 is not a tumor suppressor gene target in head and neck cancer. Genes Chromosomes Cancer 27:319-322, 2000. PMID- 10679923 TI - Recurrent allelic deletions of chromosome arms 15q and 16q in human small cell lung carcinomas. AB - The genetic lesions that lead to the development of small cell lung carcinoma (SCLC) remain incompletely defined. To identify recurrent allelic deletions in specific chromosomal regions that could serve as markers for tumor suppressor gene (TSG) inactivation in SCLC, we performed a comprehensive allelotype analysis of all 39 nonacrocentric autosomal arms. Alterations in 158 polymorphic microsatellite alleles were examined in 24 pairs of human SCLC tumor and normal control DNA samples. A total of 2,107 informative reactions were analyzed. This analysis revealed allelic losses of 100% on chromosome arm 3p, >85% loss within chromosome arms 13q and 17p, and >70% loss within chromosome arms 4q, 5q, 15q, and 16q. The allelic deletions on chromosome arms 15q and 16q have not been defined previously for SCLC and are candidate regions to harbor novel TSGs. Genes Chromosomes Cancer 27:323-331, 2000. PMID- 10679924 TI - PRK, a cell cycle gene localized to 8p21, is downregulated in head and neck cancer. AB - The human PRK gene encodes a protein serine/threonine kinase of the polo family and plays an essential role in regulating meiosis and mitosis. We have previously shown that PRK expression is downregulated in a significant fraction of lung carcinomas. Our current studies reveal that PRK mRNA expression is downregulated in a majority (26 out of 35 patients) of primary head and neck squamous-cell carcinomas (HNSCC) compared with adjacent uninvolved tissues from the same patients, regardless of stage. In addition, PRK transcripts were undetectable in one of the two HNSCC cell lines analyzed. Ectopic expression of PRK, but not a PRK deletion construct, in transformed A549 fibroblast cells suppresses their proliferation. Furthermore, fluorescence in situ hybridization analyses show that the PRK gene localizes to chromosome band 8p21, a region that exhibits a high frequency of loss of heterozygosity in a variety of human cancers, including head and neck cancers, and that is proposed to contain two putative tumor suppressor genes. Considering that PRK plays an important role in the regulation of the G2/M transition and cell cycle progression, our current studies suggest that deregulated expression of PRK may contribute to tumor development. Genes Chromosomes Cancer 27:332-336, 2000. PMID- 10679925 TI - Expression of the imprinted genes MEST/Mest in human and murine placenta suggests a role in angiogenesis. AB - In the mouse fetus, Mest is widely expressed in mesoderm derived tissues. In separate studies in mice and in humans, it has been shown to be maternally imprinted, that is, only the paternally inherited allele is active. Here, we show that starting with implantation, Mest is also expressed in maternal decidua of the mouse and in placenta of both humans and mice. Expression in murine decidua was restricted to endothelial cells. After Day 7, expression in the decidua gradually decreased. Mest-specific RT-PCR and restriction fragment length variant (RFLV) analysis of decidualized endometrium isolated from (M. musculus x M. spretus)F1 females showed that only the paternally derived Mest allele was activated in the decidual endothelium. In the mouse extraembryonic tissues, Mest transcripts were detected in derivatives of extraembryonic mesoderm only. Here, hemangioblast precursor cells and endothelial cells were positive. At all developmental stages of the mouse, trophoblast-derived cells were clearly devoid of Mest transcripts. In the human placenta MEST transcripts were also detected in hemangioblast precursor cells, however, MEST was also expressed in villous and invasive cytotrophoblast. In a human choriocarcinoma/trophoblastic tumour grown in a nude mouse, human MEST was expressed in the tumour cells, whereas murine Mest was expressed in endothelia of the murine capillaries. The expression pattern exhibited by both Mest and MEST in extraembryonic tissues during development and during formation of choriocarcinoma/trophoblast tumour suggests a functional role of the MEST proteins related to oncofetal angiogenesis. Dev Dyn 2000;217:1-10. PMID- 10679926 TI - Embryonic vasculogenesis by endothelial precursor cells derived from lung mesenchyme. AB - During development, the lung mesenchyme has a dynamic relationship with the branching airway. Embryonic lung mesenchyme is loosely packed and composed of indistinguishable cells, yet it is the source of vascular progenitors that will become endothelial cells, smooth muscle cells and fibroblasts. In the lung, vessel development in the periphery proceeds first through vasculogenesis, the migration and assembly of cells into a primitive network, and subsequently, through angiogenesis, the sprouting of vessels from this network. As a way to assess the cellular and molecular mechanisms of lung vascularization, we have isolated and cloned cell lines from mouse fetal lung mesenchyme (MFLM). Two of these MFLM cell lines, MFLM-4 and MFLM-91U, display characteristics of an endothelial lineage. RNA analysis demonstrates transcripts for the vascular endothelial growth factor receptors R1 and R2, the receptor tyrosine kinases, Tie 1 and Tie-2, as well as the Tie-2 ligands, Ang-1 and -2. The MFLM cell lines form extensive networks of capillary-like structures with lumens when cultured on a reconstituted basement membrane. In vivo, following blastocyst injection, the MFLM cells chimerize endothelium of the lung and areas of the heart vasculature. The results from these studies suggest that MFLM-4 and MFLM-91U, derived from embryonic lung mesenchyme, can function in vitro and in vivo as endothelial precursors and as models of cardiopulmonary vascularization. Dev Dyn 2000;217:11 23. PMID- 10679927 TI - Regulation of FGF soluble receptor type 1 (SR1) expression and distribution in developing, degenerating, and FGF2-treated retina. AB - The spatial and temporal patterns of expression and content of the fibroblast growth factor (FGF) soluble receptor SR1, a specific inhibitor of FGF, were investigated during embryonic and postnatal development of the retina in Fisher rats. As early as at embryonic day 18 (E18), SR1 mRNA and protein were detected in the retina. SR1 protein was strongly associated with the differentiating ganglion cells and its distribution paralleled the radial pattern of retinal development, from center to periphery. From E18 to postnatal day 5, the levels of both SR1 mRNA and SR1 protein remained constant. Thereafter, they decreased rapidly, by a factor of 5 in the adult retina. SR1 was labeled in the inner nuclear layer, but never in the photoreceptor nuclei. In the neural retina of RCS dystrophic rats, the levels SR1 mRNA and SR1 protein were 2 to 3 times higher than those in the normal congenic controls, before and during photoreceptor degeneration. These results provide the first evidence that a natural FGF inhibitor is regulated during retina development and degeneration and suggest that changes in SR1 content may be involved in the regulation of FGF activities in retina. This was confirmed in vivo in RCS rats, in which delayed photoreceptor apoptosis by intravitreal injection of FGF2 was accompanied by a downregulation of SR1 expression. Dev Dyn 2000;217:24-36. PMID- 10679928 TI - Evidence for the expression of neonatal skeletal myosin heavy chain in primary myocardium and cardiac conduction tissue in the developing chick heart. AB - We isolated a neonatal skeletal myosin heavy chain (MHC) cDNA clone, CV11E1, from a cDNA library of embryonic chick ventricle. At early cardiogenesis, diffuse expression of neonatal skeletal MHC mRNA was first detected in the heart tube at stage 10. During subsequent embryonic stages, the expression of the mRNA in the atrium was upregulated until shortly after birth. It then diminished, dramatically, and disappeared in the adult. On the other hand, in the ventricle, only a trace of the expression was detected throughout embryonic life and in the adult. However, transient expression of mRNA in the ventricle was observed, post hatching. At the protein level, during the embryonic stage, the atrial myocardium was stained diffusely with monoclonal antibody 2E9, specific for chick neonatal skeletal MHC, whereas the ventricles showed weak reactivity with 2E9. At the late embryonic and newly hatched stages, 2E9-positive cells were located clearly in the subendocardial layer, and around the blood vessels of the atrial and ventricular myocardium. These results provide the first evidence that the neonatal skeletal MHC gene is expressed in developing chick hearts. This MHC appears during early cardiogenesis and is then localized in cardiac conduction cells. Dev Dyn 2000;217:37-49. PMID- 10679929 TI - Repression of myosin isoforms in developing and denervated skeletal muscle fibers originates near motor endplates. AB - During development of chicken pectoralis muscle, a neonatal myosin heavy-chain isoform is supplanted progressively by an adult isoform. This expression is under neuronal control. In this study we test the hypothesis that developmental myosin transformations are initiated near the motor endplate of each muscle fiber, thereafter progressing toward the fiber ends. By using immunocytochemical methods, pectoralis muscle from chickens aged 1-115 days after hatching were labeled by antibody against neonatal isoform. Ellipse minor axis and mean optical density of labeled and/or unlabeled fiber profiles from each bird were measured by computer image analysis. Acetylcholinesterase (AChE) activity was demonstrated histochemically. Using serial cross sections, we show that smaller fiber profiles are the tapered ends of larger fiber profiles. The largest fiber profiles (central regions of the fibers) were the first to lose their neonatal myosin during development. Motor endplates were localized by AChE activity to the central regions of the fibers. The pectoralis of mature chickens was denervated for 3, 7, 15, or 21 days. After 2 weeks' denervation, neonatal myosin is first reexpressed in the fiber ends. Dev Dyn 2000;217:50-61. PMID- 10679930 TI - Regulation of the Hoxa4 and Hoxa5 genes in the embryonic mouse lung by retinoic acid and TGFbeta1: implications for lung development and patterning. AB - We have previously described a 5; cis-acting retinoic acid response element that is required for a subset of Hoxa4 expression, including the midgestation mouse lung. As both retinoids and Hox genes have been implicated in lung development and patterning, we have examined Hoxa4 expression in the developing mouse lung and extended our work on its regulation. At E12.5, a Hoxa4/lacZ transgene is expressed in the mesenchymal compartment of the lung. Later in development expression is restricted to the proximal mesenchyme and is also observed in smooth muscle cells, subepithelial fibroblasts, and alveolar cells. We show that both Hoxa4 and Hoxa5 are upregulated when cultured in the presence of all-trans retinoic acid. In addition, retinoic acid extends the domain of Hoxa4 and Hoxa5 expression to the periphery of the explants where the distal epithelia are developing. Interestingly, the effect of retinoic acid on Hoxa5 expression was not observed in a Hoxa4 mutant background. In contrast, TGFbeta1 was found to downregulate both Hoxa4 and Hoxa5 expression in cultured lung explants. We also establish that retinoic acid has the effect of proximalizing the mouse lung when cultured in a serum-free medium, as evidenced by reduced expression of the distal marker surfactant protein-C. Lungs from Hoxa4 mutant embryos exhibited a similar response to retinoic acid, suggesting that Hoxa4 alone is not required for the proximalizing effect. Based on their retinoid-dependent expression, we conclude that members of the group 4 and/or group 5 Hox genes are likely to be involved in patterning of the mouse lung. Dev Dyn 2000;217:62-74. PMID- 10679931 TI - Suppression of atrial myosin gene expression occurs independently in the left and right ventricles of the developing mouse heart. AB - Many cardiac genes are broadly expressed in the early heart and become restricted to the atria or ventricles as development proceeds. Additional transcriptional differences between left and right compartments of the embryonic heart have been described recently, in particular for a number of transgenes containing cardiac regulatory elements. We now demonstrate that three myosin genes which become transcriptionally restricted to the atria between embryonic day (E) 12.5 and birth, alpha-myosin heavy chain (MHC), myosin light chain (MLC) 1A and MLC2A, are coordinately downregulated in the compact myocardium of the left ventricle before that of the right ventricle. alpha-MHC protein also accumulates in the right, but not left, compact ventricular myocardium during this period, suggesting that this transient regionalization contributes to fktal heart function. dHAND and eHAND, basic helix-loop-helix transcription factors known to be expressed in the right and left ventricles respectively at E10. 5, remain regionalized between E12.5 and E14.5. Downregulation of alpha-MHC, MLC1A, and MLC2A in iv/iv embryos, which have defective left/right patterning, initiates in the morphological left (systemic) ventricle regardless of its anatomical position on the right or left hand side of the heart. This points to the importance of left/right ventricular differences in sarcomeric gene expression patterns during fktal cardiogenesis and indicates that these differences originate in the embryo in response to anterior-posterior patterning of the heart tube rather than as a result of cardiac looping. Dev Dyn 2000;217:75-85. PMID- 10679932 TI - Differential expression of endothelial beta-catenin and plakoglobin during development and maturation of the blood-brain and blood-retina barrier in the chicken. AB - The development of the blood-brain barrier depends upon the formation of a closely regulated system of adherens and tight junctions. A prerequisite for a functional junction system is the linkage of transmembrane adhesion receptors (cadherins) to the cytoskeleton via catenins. The localization of specific catenins at the adherens junction correlates with the stability of interendothelial contacts in vitro, but in vivo data are lacking thus far. Investigating brain angiogenesis in the chicken, we demonstrated that beta catenin, but not plakoglobin, initially codistributed with N-cadherin at the ablumenal endothelial membrane at contact sites to perivascular cells, from where both antigens disappeared during blood-brain barrier maturation. In contrast, plakoglobin was most prominent at the interendothelial junction where only small amounts of beta-catenin were present. Western-blot analysis revealed a stronger developmental decrease of beta-catenin than plakoglobin, whereas N-cadherin was completely lost. beta-Catenin but not N-cadherin was reinduced in brain endothelial cells during dedifferentiation in vitro and localized to the interendothelial junctions. These first in vivo data support the hypothesis that endothelial beta-catenin and N-cadherin are transiently relevant for the contact of brain endothelial to perivascular cells. Plakoglobin seems not to interact with N-cadherin but is exclusively localized at interendothelial junctions providing evidence for its role in the formation of stable adherens junctions, which may play a role for the initiation, and/or stabilization of tight junctions. Dev Dyn 2000;217:86-98. PMID- 10679933 TI - Filamin isogene expression during mouse myogenesis. AB - The developmental pattern of filamin gene expression has been studied in mouse embryos by using in situ hybridization. The probes used were isoform specific, (35)S-labeled antisense complementary ribonucleic acids (cRNAs) to the 3; untranslated region (3; UTR) of muscle-specific and nonmuscle-specific filamin genes. Northern blot and in situ hybridization results showed that nonmuscle specific filamin transcripts had a size of 9.5 kb and were expressed in all nonmuscle tissues. Labeling was most intense in tissues containing a substantial proportion of epithelial and smooth muscle cells. Muscle-specific filamin transcripts had a size of 10 kb and were expressed primarily in cardiac and skeletal muscle. The expression of muscle-specific filamin messenger ribonucleicacids (mRNAs) was detected in heart at 8.0 days after coitum, whereas that in the myotomes of somites was not detected until 10.5 days after coitum. The expression of muscle-specific filamin mRNAs in heart and in skeletal muscle continued through the subsequent days of myogenesis. The results showed that muscle-specific filamin gene transcripts are detected before the formation of myotubes in vivo. This is the first study of filamin gene expression at the early stages of skeletal muscle development. Dev Dyn 2000;217:99-108. PMID- 10679934 TI - A novel pbx family member expressed during early zebrafish embryogenesis forms trimeric complexes with Meis3 and Hoxb1b. AB - pbx genes encode homeodomain-containing transcriptional regulators that interact with other proteins to control embryogenesis and tumorigenesis. We present the characterization of a zebrafish pbx CDNA that appears to encode a novel family member, pbx4. pbx4 RNA is maternally deposited and is detected throughout the zebrafish embryo during blastula stages. It becomes excluded from ventroanterior structures at late gastrula stages and is detected within the forming central nervous system during segmentation stages. pbx4 expression overlaps with that of two other homeobox genes, hoxb1b and meis3, in the region of the presumptive caudal hindbrain during gastrula stages. In vitro binding experiments revealed that protein complexes containing Pbx4/Meis3 and Pbx4/Hoxb1b, but not Meis3/Hoxb1b could be generated. A novel trimeric complex containing Pbx4, Meis3, and Hoxb1b was also formed. We speculate that complexes with different combinations of Pbx4, Meis3, and Hoxb1b specify different developmental fates during vertebrate embryogenesis. Dev Dyn 2000;217:109-119. PMID- 10679935 TI - Transgenic rescue of aganglionosis and piebaldism in lethal spotted mice. AB - Complete colonization of the gut by enteric neural precursors depends on activation of ednrB and Ret receptors by their respective ligands, edn3 and gdnf. Mutations that eliminate expression of either ligand or either receptor produce intestinal aganglionosis in rodents and humans. Embryos homozygous for the lethal spotted (ls) allele, a loss of function mutation in the edn3 gene, have no ganglion cells in their terminal large intestines and are spotted, due to incomplete colonization of the skin by melanocyte precursors. Expression of edn3 in enteric neural precursors of transgenic mice compensates fully for deficient endogenous edn3 in ls/ls embryos. The effects of the edn3 transgene are dose dependent, as lower levels of expression in one line prevent aganglionosis in only a subset of animals and reduce, but fail to eliminate, piebaldism. In contrast, expression of neither constitutively active Ret nor activated ras in enteric neural progenitors alters the severity of aganglionosis or piebaldism in ls/ls mice. Given the spatial and temporal pattern of edn3-transgene expression, our results suggest that edn3/ednrB signals are not required prior to the arrival of crest cells in the gut and endrB stimulation elicits distinct cellular responses from Ret or ras activation. Dev Dyn 2000;217:120-132. PMID- 10679936 TI - Mutations in the X-linked pyruvate dehydrogenase (E1) alpha subunit gene (PDHA1) in patients with a pyruvate dehydrogenase complex deficiency. AB - Defects in the pyruvate dehydrogenase (PDH) complex are an important cause of primary lactic acidosis, a frequent manifestation of metabolic disease in children. Clinical symptoms can vary considerably in patients with PDH complex deficiencies, and almost equal numbers of affected males and females have been identified, suggesting an autosomal recessive mode of inheritance of the disease. However, the great majority of PDH complex deficiencies result from mutations in the X-linked pyruvate dehydrogenase (E1) alpha subunit gene (PDHA1). The major factors that contribute to the clinical variation in E1alpha deficiency and its resemblance to a recessive disease are developmental lethality in some males with severe mutations and the pattern of X-inactivation in females. To date, 37 different missense/nonsense and 39 different insertion/deletion mutations have been identified in the E1alpha subunit gene of 130 patients (61 females and 69 males) from 123 unrelated families. Insertion/deletion mutations occur preferentially in exons 10 and 11, while missense/nonsense mutations are found in all exons. In males, the majority of missense/nonsense mutations are found in exons 3, 7, 8 and 11, and three recurrent mutations at codons R72, R263 and R378 account for half of these patients with missense/nonsense mutations (25 of 50). A significantly lower number of females is found with missense/nonsense mutations (25). However, 36 females out of 55 affected patients have insertion/deletion mutations. The total number of female and male patients is thus almost the same, although a difference in the distribution of the type of mutations is evident between both sexes. In many families, the parents of the affected patients were studied for the presence of the PDHA1 mutation. The mutation was never present in the somatic cells of the father; in 63 mothers studied, 16 were carriers (25%). In four families, the origin of the new mutation was determined to be twice paternal and twice maternal. PMID- 10679937 TI - Molecular basis of multiple exostoses: mutations in the EXT1 and EXT2 genes. AB - Hereditary multiple exostoses (EXT) is an autosomal dominant disorder characterized by the formation of exostoses, which are cartilage-capped bony protuberances mainly located on long bones. Two genes, EXT1 and EXT2, and at least one other unidentified gene, are known to be involved in the formation of exostoses. To date, 49 different EXT1 and 25 different EXT2 mutations have been found in EXT patients, and there is evidence that mutations in these two genes are responsible for over 70% of the EXT cases. Among the 49 EXT1 mutations there are 9 nonsense, 21 frameshift, and 5 splice site mutations; 2 in-frame deletions of 1 and 5 amino acids respectively; and 12 missense mutations. For EXT2, 8 nonsense, 11 frameshift, 3 splice site and 3 missense mutations are described. The majority of these mutations are mutations causing loss of function, which is consistent with the presumed tumor suppressor function of the EXT genes. PMID- 10679938 TI - An update of the mutation spectrum of the survival motor neuron gene (SMN1) in autosomal recessive spinal muscular atrophy (SMA). AB - Spinal muscular atrophy (SMA) is characterized by degeneration of motor neurons in the spinal cord, causing progressive weakness of the limbs and trunk, followed by muscle atrophy. SMA is one of the most frequent autosomal recessive diseases, with a carrier frequency of 1 in 50 and the most common genetic cause of childhood mortality. The phenotype is extremely variable, and patients have been classified in type I-III SMA based on age at onset and clinical course. All three types of SMA are caused by mutations in the survival motor neuron gene (SMN1). There are two almost identical copies, SMN1 and SMN2, present on chromosome 5q13. Only homozygous absence of SMN1 is responsible for SMA, while homozygous absence of SMN2, found in about 5% of controls, has no clinical phenotype. Ninety-six percent of SMA patients display mutations in SMN1, while 4% are unlinked to 5q13. Of the 5q13-linked SMA patients, 96.4% show homozygous absence of SMN1 exons 7 and 8 or exon 7 only, whereas 3. 6% present a compound heterozygosity with a subtle mutation on one chromosome and a deletion/gene conversion on the other chromosome. Among the 23 different subtle mutations described so far, the Y272C missense mutation is the most frequent one, at 20%. Given this uniform mutation spectrum, direct molecular genetic testing is an easy and rapid analysis for most of the SMA patients. Direct testing of heterozygotes, while not trivial, is compromised by the presence of two SMN1 copies per chromosome in about 4% of individuals. The number of SMN2 copies modulates the SMA phenotype. Nevertheless, it should not be used for prediction of severity of the SMA. PMID- 10679939 TI - A missense mutation in the OCTN2 gene associated with residual carnitine transport activity. AB - Primary carnitine deficiency is an autosomal recessive disorder of fatty acid oxidation caused by defective carnitine transport. This disease can present early in life with hypoketotic hypoglycemia and acute metabolic decompensation, or later in life with skeletal or cardiac myopathy. Mutations abolishing the function of OCTN2, an organic cation/carnitine transporter with twelve putative transmembrane spanning domains, were recently demonstrated in patients with early and late-onset (up to seven years of age) presentation of this syndrome. Most of the reported mutations are null alleles. Here we evaluate the OCTN2 gene in a male patient who presented at seven years of age with severe dilated cardiomyopathy. Plasma carnitine levels were undetectable and carnitine transport by his fibroblasts was reduced to about 3% of normal controls. This patient was homozygous for a single base pair change in exon 8 of the OCTN2 gene (1354G>A) converting the codon for Glu 452 to Lys (E452K) in the predicted intracellular loop between transmembrane domains 10 and 11. Stable expression of the mutant E452K-OCTN2 cDNA in Chinese hamster ovary (CHO) cells caused a partial increase in carnitine transport to 2-4% of the levels measured in the wild type transporter. This reduced transport activity was associated with normal Km toward carnitine (3.1 +/- 1.1 microM), but markedly reduced Vmax. These results indicate that primary carnitine deficiency can be caused by mutations encoding for carnitine transporters with residual activity, and that the E452K affects a domain not involved in carnitine recognition. PMID- 10679940 TI - Screening of thiopurine S-methyltransferase mutations by horizontal conformation sensitive gel electrophoresis. AB - The genetic polymorphism of thiopurine S-methyltransferase (TPMT) has had a highly significant clinical impact due to its association with individual variation in the toxicity and therapeutic efficiency of thiopurine drugs, which are pharmaceutical agents widely used in the treatment of several kinds of diseases. Until now, ten mutant alleles responsible for TPMT deficiency and several silent and intronic mutations have been described. In this work we present an alternative molecular method for the detection of TPMT alleles. It is an adaptation for horizontal conditions of a conformation-sensitive gel electrophoresis technique. The method has proven to be very efficient as a rapid screening approach for the study of TPMT genetic variability. The method was applied to analyse eight TPMT exons and the corresponding flanking intronic regions in a sample of unrelated healthy individuals from North Portugal. Here we report the allelic frequencies concerning TPMT-deficient alleles and several silent and intronic mutations, including two newly detected intronic polymorphisms: an A (-101) T substitution in intron 3 and a variation involving the number of T nucleotides in a DNA stretch in intron 5. Additionally, we also present data from a sample of 43 children undergoing therapy for acute lymphoblastic leukemia. In this clinical sample we have registered a statistically significant higher frequency for the TPMT*3C allele. This finding raises the question whether the TPMT genotype can contribute to any genetic predisposition for development of the malignancy. PMID- 10679941 TI - Phenylketonuria and hyperphenylalaninemia in eastern Germany: a characteristic molecular profile and 15 novel mutations. AB - Phenylketonuria (PKU) is an important error of amino acid metabolism which results in most patients from phenylalanine hydroxylase (PAH) deficiency. PKU displays a marked genotypic heterogeneity both within and between different populations. The aim of this study was to establish the genotypic spectrum of PKU in eastern Germany, and to compare this to the distribution of mutations in western Germany. The study population included 302 patients in 290 families who were followed at treatment centers in Berlin, Leipzig and Jena. The study showed marked genotypic variability with a total of 75 mutations, including 15 that have so far not been described (eleven missense mutations, one splicing mutation, and three small deletions). One of these novel mutations, E183Q, occurred in cis to a R408W mutation. In the non-immigrant eastern German population, the frequency of R408W accounted for 40.1% of the PKU alleles. In the immigrant Turkish population of the former West Berlin, the most prevalent mutation was IVS10-11G>A (57%). There was a marked difference of the genotypic spectrum between the population studied here and the data reported from the western part of the country. PMID- 10679942 TI - Eight novel mutations and consequences on mRNA and protein level in pyruvate kinase-deficient patients with nonspherocytic hemolytic anemia. AB - Pyruvate kinase (PK) deficiency (PKD) is an autosomal recessive disorder with the typical manifestation of nonspherocytic hemolytic anemia. We analyzed the mutant enzymes of 10 unrelated patients with PKD, whose symptoms ranged from a mild, chronic hemolytic anemia to a severe anemia, by sequence analysis for the presence of alterations in the PKLR gene. In all cases the patients were shown to be compound heterozygous. Eight novel mutations were identified: 458T-->C (Ile153Thr), 656T-->C (Ile219Thr), 877G-->A (Asp293Asn), 991G-->A (Asp331Asn), 1055C-->A (Ala352Asp), 1483G-->A (Ala495Thr), 1649A-->T (Asp550Val), and 183 184ins16bp. This 16 bp duplication produces a frameshift and subsequent stop codon resulting in a drastically reduced mRNA level, and probably in an unstable gene product. Surprisingly, the existence of M2-type PK could be demonstrated in the patient's red blood cells. The study of different polymorphic sites revealed, with one exception, a strict linkage of the 1705C, 1738T, IVS5(+51)T, T(10) polymorphisms and the presence of 14 ATT repeats in intron 11. Our analyses show the consequences of a distorted structure on enzyme function and we discuss the correlations between the mutations identified and the parameters indicative for enzyme function. PMID- 10679943 TI - Detection of eight novel palmitoyl protein thioesterase (PPT) mutations underlying infantile neuronal ceroid lipofuscinosis (INCL;CLN1). AB - The infantile form of neuronal ceroid lipofuscinosis (INCL; CLN1) is the earliest onset form of the neuronal ceroid lipofuscinoses (NCL), a group of progressive encephalopathies of children. INCL is caused by mutations in the palmitoyl protein thioesterase (PPT) gene, and we report here eight novel INCL mutations in PPT. Five of the mutations, c.456C>A, c.162-163insA, c.174-175delG, c.774 775insA, and a splice acceptor mutation IVS1-2A>G in intron 1, caused the generation of a premature STOP codon either directly or after a frameshift. One mutation was a three-bp insertion in exon 2 (c. 132-133insTGT) leading to insertion of one extra cysteine (Ser44-insCys-Cys45), and another mutation, a 3 bp deletion in exon 3 (c.249-251delCTT), led to deletion of Phe84 in PPT. A splice acceptor mutation IVS6-1G>T in intron 6 can be predicted to cause skipping of exon 7 in PPT. All of these novel mutations were associated with the classical phenotype of INCL, with the first symptoms starting around 12 months of age. The severe phenotypes could be explained by the nature of the novel mutations: five are predicted to lead to premature translational termination, thus abolishing the active site of PPT, and three will probably cause a misfolding of the nascent polypeptide. Thirty-five percent (7/20) of the disease alleles in these 11 families contained the most prevalent c.451C>T missense mutation outside Finland [Das et al., 1998]. Consequently, 31 different mutations underlying INCL have been found so far, the majority leading to classical INCL. PMID- 10679944 TI - Characterization of six novel mutations in the methylenetetrahydrofolate reductase (MTHFR) gene in patients with homocystinuria. AB - Severe deficiency of methylenetetrahydrofolate reductase (MTHFR) is the most common inborn error of folate metabolism. Patients are characterized by severe hyperhomocysteinemia, homocystinuria and a variety of neurological and vascular problems. Eighteen rare mutations have been reported in this group of patients. Two polymorphisms which cause mild enzyme deficiencies have been described (677C- >T and 1298A-->C). The first sequence change encodes a thermolabile enzyme and is associated with mild hyperhomocysteinemia. Six novel point mutations are described in patients with severe deficiency of MTHFR, along with their associated polymorphisms and clinical phenotypes. Of the two nonsense mutations (1762A-->T, 1134C-->G) and four missense mutations (1727C-->T, 1172G-->A, 1768G- >A, and 358G-->A), one was identified in the N-terminal catalytic domain, while the others were located in the regulatory C-terminal region. All four residues affected by missense mutations are conserved in one or more MTHFRs of other species. This report brings the total to 24 mutations identified in severe MTHFR deficiency, with two mutations identified in each of 22 patients. PMID- 10679946 TI - Fifteen new mutations (-195C>T, L-12X, 298-2A>G, T117N, A159T, R229S, 997+2T>A, E274X, A331T, H364R, D389G, 1256delC, R433H, N461I, C472S) in the tissue nonspecific alkaline phosphatase (TNSALP) gene in patients with hypophosphatasia. AB - Hypophosphatasia is a rare inherited disorder characterized by defective bone mineralization and deficiency of serum and liver/bone/kidney-type alkaline phosphatase (L/B/K ALP) activity. We report the characterization of tissue nonspecific alkaline phosphatase (TNSALP) gene mutations in a series of 12 families affected by severe or mild hypophosphatasia. Twenty distinct mutations were found, 5 of which were previously reported. Nine of the 15 new mutations were missense mutations (T117N, A159T, R229S, A331T, H364R, D389G, R433H, N461I, and C472S). The others were 2 nonsense mutations (L-12X and E274X), one single nucleotide deletion (1256delC), 2 mutations affecting splicing (298-2A>G, 997+2T>A), and a mutation in the major transcription start site (-195C>T). Hum Mutat 15:293, 2000. PMID- 10679947 TI - Mutation 985A>G in the MCAD gene shows low incidence in Estonian population. AB - Medium-chain acyl-CoA dehydrogenase (MCAD) is an enzyme responsible for large part of mitochondrial beta-oxidation of fatty acids and therefore stays on key position of cellular energy supply. In case of its deficiency, starvation, rapid growth periods or infections may cause fatal lack of energy, especially in the first years of life. MCAD deficiency is inherited in an autosomal recessive manner and it has been shown to be rather common in some European countries (Great Britain 1 in 6,000, Switzerland 1 in 10,000). In Caucasoid populations one mutation, the 985A>G transition, causing the amino acid substitution K329E, accounts for about 90% of all mutant MCAD alleles. Here we present data about screening the Estonian population for this mutation. We analyzed the DNA from 1,098 persons from all regions of Estonia (all newborns born in one month) and found 5 heterozygotes for 985A>G, that makes the carrier frequency 1 in 220 and the frequency of possibly affected homozygotes 1 out of 193,000. No mutant alleles were found among the samples of the children, who had unclear diagnosis for death during the years 1994 and 1995. PMID- 10679948 TI - Two homozygous mutations (R193W and 794/795 delAA) in the myophosphorylase gene in a patient with McArdle's disease. AB - We report two novel homozygous mutations in the myophosphorylase gene (PYGM) in a patient with McArdle's disease. A C-to-T transition that changed an arginine to tryptophan at codon 193 (R193W) in exon 5, and a deletion of two adenine base pairs in exon 20 at codon 794/795 (794/795 delAA) were identified. Several lines of evidence suggest the pathogenicity of both mutations: (i) they were the only nucleotide alteration in the coding region and adjacent exon/intron boundaries of the PYGM gene; (ii) the R193W mutation leads to the replacement of a highly conserved amino acid residue involved in glucose-6-P binding, and the 794/795 delAA mutation predicts a frameshift and premature termination of the protein; (iii) 60 normal controls and 20 disease controls did not have the mutations in their 160 alleles. Hum Mutat 15:294, 2000. PMID- 10679949 TI - Microsomal triglyceride transfer protein (MTP) gene mutations in Canadian subjects with abetalipoproteinemia. AB - Abetalipoproteinemia (ABL) is an extremely rare autosomal recessive disorder, which is characterized by defective assembly and secretion of plasma apolipoprotein (apo) B-containing lipoproteins. ABL results from mutations in the gene encoding the microsomal triglyceride transfer protein (MTP). We sequenced the MTP gene in six Canadian subjects with ABL, of whom four were found to be simple homozygotes and two were found to be compound heterozygotes for MTP gene mutations. Of the 8 MTP gene mutations identified, 6 had not been previously reported, including two new nonsense mutations (K448X and K842X), two new missense mutations (S590I and G746E), one new frameshift mutation (1820del1) and one new splice donor site mutation (G1770A). Despite appropriate treatment with high doses of fat-soluble vitamins in all subjects, there was a wide variation in the progression and severity of the clinical phenotypes. For example, the presence of severe retinopathy and neuropathy did not correlate with the type and position of the mutation, but rather with the age at diagnosis and onset of treatment with fat-soluble vitamins. These findings suggest that genetic and non genetic factors can modulate the clinical impact of mutant MTP in ABL patients. PMID- 10679950 TI - High incidence of 550delA mutation of CAPN3 in LGMD2 patients from Russia. AB - Autosomal recessive limb gird muscular dystrophy (LGMD2) is a clinically and genetically heterogeneous group of diseases that are characterized by progressive atrophy and weakness of the proximal limb muscles. At least eight genetic loci leading to LGMD2 are recognized. The proportion of particular gene involved in producing different forms of LGMD2 shows a marked geographical variation. We studied 19 LGMD2 patients from Russia (15 families) and found calpain 3 (CAPN3) gene mutations in most of the patients studied. Sequence analysis of the fourth exons revealed two sibs - heterozygous compound for a 15-bp deletion (nt598-612) and 550 adenine deletion, and two sibs homozygous for a 550delA. We developed assay based on allele specific amplification (ASA) for rapid screening of the 550delA. The ASA assay of the LGMD2 patients under study showed that 7 patients from 6 families were homozygous for 550delA and 7 patients from 4 families were heterozygous for 550delA. A linkage analysis employing four microsatellites flanking the LGMD2A locus was performed. We found complete haplotype identity in most cases what favors the possibility of a common founder. Heterozygous carriers of 550delA were found in general population. The crude estimate of the mutation frequency is 1/150. Hum Mutat 15:295, 2000. PMID- 10679951 TI - IDDM7 links to insulin-dependent diabetes mellitus in Danish multiplex families but linkage is not explained by novel polymorphisms in the candidate gene GALNT3. The Danish Study Group of Diabetes in Childhood and The Danish IDDM Epidemiology and Genetics Group. AB - The insulin-dependent diabetes mellitus (IDDM) susceptibility locus IDDM7 on 2q31 links to IDDM in some but not other populations. Linkage of D2S152, the marker for IDDM7, has hitherto not been demonstrated in Danish patients. GALNT3 that encodes the UDP-GalNAc: polypeptide N-acetyl-galactosaminyltransferase-T3 (GalNAc T3), was recently identified and mapped to a region 5-25 cM from D2S152. The GalNAc transferases may play a role in immune mediated diseases by glycosylating autoantigens. Hence, the aims of the present study were to investigate by means of extended transmission disequilibrium testing (ETDT) and transmission disequilibrium testing (TDT) of the marker for IDDM7, D2S152, the marker for GALNT3, D2S2363, and novel polymorphisms identified through mutation screening of the entire GALNT3 for linkage with IDDM in 241 Danish IDDM multiplex families. ETDT analysis demonstrated linkage between IDDM and D2S152 (P(ETDT)=0.034). A prevalent T-->A polymorphism, T284A, was found in the GALNT3 3'UTR. Analysis of the D2S2363 and the T284A GALNT3 transmission patterns did not show linkage to IDDM in Danish patients (P(ETDT)=0.15 and P(TDT)=0.76, respectively). In conclusion, IDDM7 (D2S152) links to IDDM in Danish patients, but D2S2363 and the identified T284A polymorphism in the GALNT3 3'UTR did not. Hence, it is unlikely that the GALNT3 is an IDDM susceptibility gene. PMID- 10679952 TI - PCR diagnosis of X-linked ichthyosis: identification of a novel mutation (E560P) of the steroid sulfatase gene. AB - X-linked ichthyosis (XLI) is an inherited skin disorder due to deficiency of steroid sulfatase (STS) activity. XLI has been diagnosed by assaying STS activity in placenta or lymphocytes of patients after birth. Most patients have a large deletion of the STS gene, generated by inaccurate recombination at the STS locus. However, point mutations in the STS gene have been reported in some patients with complete STS deficiency. In a new case of STS deficiency, we identified an STS missense mutation, Glu560Pro or E560P. This new point mutation suggests that the C-terminal region of the STS enzyme is important for STS enzymatic function. Hum Mutat 15:296, 2000. PMID- 10679953 TI - Various AGC repeat numbers in the coding region of the human transcription factor gene E2F-4. AB - The E2F family of transcription factors regulates the expression of genes required for DNA synthesis and cell cycle control. The AGC triplet repeat in the coding region of the E2F-4 gene, a member of the family, has been reported to be mutated in colorectal cancers with a microsatellite instability (MSI) phenotype. We found a wider range variation of the repeat number in DNAs from tumors, the corresponding normal mucosa, and healthy individuals. A total of 5 repeat variants, ranging from 8 to 17 AGC repeats, was detected in 6 (9.7%) of the 62 healthy individuals and 8 (8.9%) of the 90 normal DNAs of the patients. The wild type 13 repeat was present in all of these individuals. The variation of the AGC repeat number may be a polymorphism. Further, loss of heterozygosity (LOH) at the E2F-4 locus in the tumor tissues of 2 (25%) of the 8 informative cases was detected. The variation may be a useful marker for detection of LOH in primary tumors. PMID- 10679954 TI - The Arg1075His substitution in the FBN1 gene is clinically innocent for Marfan syndrome. PMID- 10679955 TI - Polymorphism (g2035C>T) in the amelogenin gene. PMID- 10679956 TI - A novel DNA polymorphism (4886C>T) in the human LCAT gene. PMID- 10679957 TI - A novel missense mutation (R712L) adjacent to the "active thiol" region of the cardiac beta-myosin heavy chain gene causing hypertrophic cardiomyopathy in an Indian family. PMID- 10679958 TI - Identification of a novel large F9 gene mutation-an insertion of an Alu repeated DNA element in exon e of the factor 9 gene. PMID- 10679959 TI - Identification of a new polymorphism (c134G>A) in the exon 2 of the myelin protein zero gene. PMID- 10679960 TI - Introduction to muscular dystrophy. PMID- 10679961 TI - Plasma membrane cytoskeleton of muscle: a fine structural analysis. AB - The discovery of dystrophin and its definition as the causative molecule in Duchenne Muscular Dystrophy has led to a renewed interest in the molecular structure of the muscle fiber plasma membrane and its association with the extracellular basal lamina. The original identification of dystrophin gave credence to the possibility that the plasma membrane of the muscle fiber may be highly organized and involved in maintaining appropriate homeostasis in this actively contracting cellular system. In this review, we examine the currently known members of the muscle fiber plasma membrane cytoskeleton and the interactions that occur between the different members of this complex using histological, electron microscopic, and confocal methods. From our studies and others cited in this review, it is clear that the dystrophin cytoskeletal complex is not completely understood and component molecules continue to be discovered. Perhaps equally importantly, currently defined molecules (such as alpha-actinin or neuronal nitric oxide synthase) are being recognized as being specifically associated with the complex. What is striking from all of the studies, to date, is that while we are able to identify members of the dystrophin cytoskeletal complex and while we are able to associate mutations of individual molecules with disease(s), we are still unable to truly define the roles of each of the molecules in maintaining the normal physiology of the muscle fiber. PMID- 10679962 TI - Immunocytochemical analysis of human muscular dystrophy. AB - Immunocytochemistry is an essential tool for the assessment of muscle biopsies from patients with muscular dystrophy, especially the recessive forms. Antibodies can detect primary defects when there is an alteration in expression, in particular in Xp21 muscular dystrophies, Emery-Dreifuss muscular dystrophy, the limb-girdle dystrophies caused by abnormal expression of the sarcoglycans, and in the form of congenital muscular dystrophy linked to the gene for laminin alpha2. Absence of a protein is easily observed and reduction in expression can be assessed provided adequate controls and baselines are established. Assessment of secondary defects can also be of diagnostic value; they widen the understanding of pathology changes, and are helping in the development of therapeutic strategies. PMID- 10679963 TI - Dystrophin and utrophin: genetic analyses of their role in skeletal muscle. AB - Since the identification of dystrophin as the causitive factor in Duchenne muscular dystrophy, there has been substantial progress in understanding the functions and interactions of this protein. Dystrophin has been shown to interact with a group of peripheral- and trans-membrane proteins known as the dystrophin associated protein complex (DAPC) and mutations in some of the members of this complex have been shown to account for other forms of muscular dystrophy. This review summarizes the experiments using transgenic and knockout mouse models that have defined the roles of dystrophin, and the dystrophin-related protein utrophin at the skeletal muscle membrane and at the neuromuscular junction. These studies are presented in the context of other known interactions at the muscle membrane. Studies of the dystrophin-deficient mdx mouse have lead to a greater understanding of the human disease. Knockouts and transgenics of utrophin have shown this protein to be sufficient to functionally compensate for dystrophin. Dystrophin transgenic mice combined with the mdx mouse have been used to study the function of specific domains of the dystrophin protein. Together these animal models have led to a delineation of protein functions and localization patterns that will be useful for the generation of potential therapies for DMD. PMID- 10679964 TI - Sarcoglycans in muscular dystrophy. AB - Muscular dystrophy is a heterogeneous genetic disease that affects skeletal and cardiac muscle. The genetic defects associated with muscular dystrophy include mutations in dystrophin and its associated glycoproteins, the sarcoglycans. Furthermore, defects in dystrophin have been shown to cause a disruption of the normal expression and localization of the sarcoglycan complex. Thus, abnormalities of sarcoglycan are a common molecular feature in a number of dystrophies. By combining biochemistry, molecular cell biology, and human and mouse genetics, a growing understanding of the sarcoglycan complex is emerging. Sarcoglycan appears to be an important, independent mediator of dystrophic pathology in both skeletal muscle and heart. The absence of sarcoglycan leads to alterations of membrane permeability and apoptosis, two shared features of a number of dystrophies. beta-sarcoglycan and delta-sarcoglycan may form the core of the sarcoglycan subcomplex with alpha- and gamma-sarcoglycan less tightly associated to this core. The relationship of epsilon-sarcoglycan to the dystrophin-glycoprotein complex remains unclear. Animals lacking alpha-, gamma- and delta-sarcoglycan have been described and provide excellent opportunities for further investigation of the function of sarcoglycan. Dystrophin with dystroglycan and laminin may be a mechanical link between the actin cytoskeleton and the extracellular matrix. By positioning itself in close proximity to dystrophin and dystroglycan, sarcoglycan may function to couple mechanical and chemical signals in striated muscle. Sarcoglycan may be an independent signaling or regulatory module whose position in the membrane is determined by dystrophin but whose function is carried out independent of the dystrophin-dystroglycan laminin axis. PMID- 10679965 TI - Merosin and congenital muscular dystrophy. AB - Merosin (also called as Laminin-2) is an isoform of laminin comprised of the alpha2, beta1 and gamma1 chains. In European populations, half of the patients with classical congenital muscular dystrophy have mutations of the LAMA2 gene (6q22-23) and present reduced or absence of laminin alpha2 chain. This form is generally referred to as merosin-deficient CMD. Merosin-deficient CMD is characterized by involvement of not only skeletal muscle but also central and peripheral nervous systems: Extensive brain white matter abnormalities are found by magnetic resonance imaging (MRI). However, most patients show no mental retardation. Recent case studies reported that some patients have several structural abnormalities such as abnormal cerebral cortical gyration, hypoplasia of cerebellum and pons, and dilation of ventricles. At present, functions of merosin related to muscle degeneration have not been fully elucidated. In addition, the mechanisms responsible for pathogenesis of diffuse brain white matter abnormalities remain to be determined. As mouse models for merosin deficient CMD, three spontaneous mutants(dy, dy(2J), dy(PAS1)) and two mutants named dy(W) and dy(3K) by targeted gene disruption have been reported. These mice will help to elucidate the pathogenesis of merosin-deficient CMD and serve to develop therapy. PMID- 10679966 TI - Eye muscle sparing by the muscular dystrophies: lessons to be learned? AB - The devastating consequences of the various muscular dystrophies are even more obvious when a muscle or muscle group is spared. The study of the exceptional cell or tissue responses may prove to be of considerable value in the analysis of disease mechanisms. The small muscles responsible for eye movements, the extraocular muscles, have functional and morphological characteristics that set them aside from other skeletal muscles. Notably, these muscles are clinically unaffected in Duchenne/Becker, limb-girdle, and congenital muscular dystrophies, pathologies due to a broken mechanical or signaling linkage between the cytoskeleton and the extracellular matrix. Uncovering the strategies used by the extraocular muscles to "naturally" protect themselves in these diseases should contribute to knowledge of both pathogenesis and treatment. We propose that careful investigation of the cellular determinants of extraocular muscle-specific properties may provide insights into how these muscles avoid or adapt to the cascade of events leading to myofiber degeneration in the muscular dystrophies. PMID- 10679967 TI - Cell cycle behavior and MyoD expression in response to T3 differ in normal and mdx dystrophic primary muscle cell cultures. AB - Since mdx limb muscle regeneration in vivo is accompanied by rapid myoblast proliferation and differentiation compared to normal, we tested the possibility that proliferation and differentiation were differentially regulated in normal and mdx dystrophic muscle cells. Cell cycle behavior, MyoD expression, and the effects of thyroid hormone (T3) treatment were examined in primary cultures. Using a 4-hour pulse time for bromodeoxyuridine (BrdU) incorporation during S phase, the phases of the cell cycle (early S, late S, G(2)/M, and G(0)/G(1)) were separated by 2-colour fluorescence (BrdU/PI) analysis using flow cytometry. The G(0)/G(1)-early S and the late S-G(2)/M transitions were examined under the influence of T3 in cycling normal and mdx muscle cell cultures over a 20-hour time period. Myogenesis and differentiation were assessed morphologically and by immunostaining for MyoD protein. Mdx cultures had fewer cells in G(0)/G(1) at 20 hours and more cells in early and late S-phase compared to normal cultures. T3 significantly increased the proportion of normal cells in early S-phase by 20 hours, and reduced the proportions in G(2)/M phase. Over the same time interval in parallel cultures, the proportion of MyoD+ normal cells decreased significantly. In the absence of T3, mdx cell cultures showed greater proportions of cells in S-phase than normal cultures, and similar increases in S-phase and loss of MyoD expression over time. However, mdx cultures had no change in the proportion that were MyoD+ during T3 treatment. The results confirm that T3 in primary cultures increased proliferation and prevented the de-differentiation of mdx cells to a greater degree than was typical of normal cells. The different susceptibilities to T3-related shifts between proliferation and differentiation observed in vitro support the idea that committed mdx myoblasts may be more activated and proliferative than normal myoblasts during regeneration in vivo. PMID- 10679968 TI - Progress in myoblast transplantation: a potential treatment of dystrophies. AB - Myoblast transplantation (MT) consists of injecting normal or genetically modified myogenic cells into muscles, where they are expected to fuse and form mature fibers. As an experimental approach to treat severe genetic muscle diseases, MT was tested in dystrophic patients at the beginning of the 1990s. Although these early clinical trials were unsuccessful, MT has progressed through the research on animal models. Many factors that may condition the success of MT were identified in the last years. The present review updates our knowledge on MT and describes the different problems that have limited its success. Factors that were first underestimated, like the specific immune response after MT, are presently well characterized. Destruction of the hybrid fibers by activated T lymphocytes and production of antibodies against the transplanted myoblasts take place after MT and are responsible for the graft rejection. The choice of the immunosuppression seems to be very important, and FK506 is the best agent known to allow the best results after MT. Under FK506 immunosuppression, very efficient MT were obtained both in mice and monkeys. Moreover, in dystrophic mice it was demonstrated that MT ameliorates some phenotypical characteristics of the disease. The improvement of the survival of the transplanted cells and the increase of their migration into the injected tissue are presently under investigation. Some of the present research is directed also to bypass the immunosuppression by using the patient's own cells for MT. In this sense, efforts are conducted to introduce the normal gene into the patient's myoblasts before MT and to improve the ability of these cells to proliferate in vitro. Micros. Res. Tech. 48:213-222, 2000. PMID- 10679970 TI - Obituary: takekiyo matsuo (1943-1999) PMID- 10679969 TI - Developments in gene therapy for muscular dystrophy. AB - Gene therapy for muscular dystrophy (MD) presents significant challenges, including the large amount of muscle tissue in the body, the large size of many genes defective in different muscular dystrophies, and the possibility of a host immune response against the therapeutic gene. Overcoming these challenges requires the development and delivery of suitable gene transfer vectors. Encouraging progress has been made in modifying adenovirus (Ad) vectors to reduce immune response and increase capacity. Recently developed gutted Ad vectors can deliver full-length dystrophin cDNA expression vectors to muscle tissue. Using muscle-specific promoters to drive dystrophin expression, a strong immune response has not been observed in mdx mice. Adeno-associated virus (AAV) vectors can deliver small genes to muscle without provocation of a significant immune response, which should allow long-term expression of several MD genes. AAV vectors have also been used to deliver sarcoglycan genes to entire muscle groups. These advances and others reviewed here suggest that barriers to gene therapy for MD are surmountable. PMID- 10679971 TI - Introduction to modern biological mass spectrometry. PMID- 10679972 TI - Structural studies of the Maillard reaction products of a protein using ion trap mass spectrometry. AB - The early stage products of the Maillard reaction of egg white lysozyme with D glucose were studied. Incubation with D-glucose at 50 degrees C for 20 days caused reaction on the Lys and Arg residues of lysozyme as follows: all of the six Lys residues and 10 of the 11 Arg residues in lysozyme reacted with D glucose; Arg 61 did not react with D-glucose. The Lys residues reacted with D glucose with 1 mol of dehydration per mole of residue, and the Arg residues reacted with 2 mol of dehydration per mole of residue. The major constituent of the Amadori product with the epsilon-amino group of the Lys residue and the D glucose was found to be the beta-pyranose form. The structure of the early stage product of the Maillard reaction of a protein with a sugar is the same as that of an amino acid with a sugar. PMID- 10679973 TI - Survey of the proton affinities of adenine, cytosine, thymine and uracil dideoxyribonucleosides, deoxyribonucleosides and ribonucleosides. AB - The kinetic method was applied to the determination of the proton affinities (PAs) of modified deoxy- and dideoxyribonucleosides. A correlation between the measured PAs and the replacement of one of the three hydroxyl groups of the ribose unit is presented. A PA scale was obtained which shows that the replacement of the primary or of one or both secondary hydroxyl groups of a ribonucleoside with a hydrogen atom induces the lowering or the enhancement of the nucleoside PA, respectively. The scale extends over a very narrow range of approximately 2 kcal mol(-1), thus demonstrating the sensitivity of the kinetic method in the evaluation of small differences in thermodynamic parameters. PMID- 10679974 TI - Structure determination of two conotoxins from Conus textile by a combination of matrix-assisted laser desorption/ionization time-of-flight and electrospray ionization mass spectrometry and biochemical methods. AB - Two highly modified conotoxins from the mollusc Conus textile, epsilon-TxIX and Gla(1)-TxVI, were characterized by matrix-assisted laser desorption/ionization and electrospray mass spectrometry and also by electrospray ionization tandem and triple mass spectrometry in combination with enzymatic cleavage and chemical modification reactions. The mass spectrometric studies allowed the confirmation of the sequence determined by Edman degradation and assignment of unidentified amino acid residues, among which bromotryptophan residues and an O-glycosylated threonine residue were observed. Methyl esterification was found necessary for the site-specific assignment of the Gla residues in the peptides. PMID- 10679975 TI - Improving the sensitivity of the end-cap reflectron time-of-flight mass spectrometer. AB - A miniaturized time-of-flight (TOF) mass spectrometer utilizes an end-cap reflectron to achieve high kinetic energy focusing and improved mass resolution. However, the coaxial geometry gives rise to considerable losses in sensitivity resulting from reflected ion trajectories close to the center. These trajectories were modeled, using initial ion velocity distributions in the radial direction up to 300 m s(-1), and the portion of the active area of the detector that is utilized was evaluated experimentally using a variable diameter iris diaphragm. The sensitivity was improved by modification of the reflectron by tilting the end cap electrode 4 degrees and redirecting the ions to a portion of the detector active area. Sensitivity was then measured as 3 fmol of the peptide substance P. PMID- 10679976 TI - Construction of a new multi-turn time-of-flight mass spectrometer. AB - A new type of multi-turn time-of-flight mass spectrometer was constructed, consisting of four cylindrical electric sectors and 28 electric quadrupole lenses, the size of the vacuum chamber being 60 x 70 x 20 cm. It was demonstrated that the mass resolution can be increased according to the number of cycles of the ions through the ion optical system. PMID- 10679977 TI - Dissociation study of tellurium cluster ions, Te(n)(+) (n = 25-85) using secondary ion mass spectrometry. AB - Dissociation pathways of tellurium clusters, Te(n)(+) (n = 25-85), were investigated by secondary ion mass spectrometry. Positively charged ions were generated from a tellurium sheet by bombardment with 10 keV xenon ion beam. Mass analyses of cluster ions were performed using a grand-scale sector mass spectrometer. In the first field-free region, Te(n)(+) (n = 25-80) had a large dissociation probability with five- and six-atom emission and Te(n)(+) (n = 50 85) had a slightly large dissociation probability with 10- and 11-atom emission. Five- and six-atom dissociation in the second field-free region could be also observed. These results were most likely due to the cluster emission processes for Te(n)(+), although sequential atom emission and cluster emission could not be distinguished by this type of experiments. PMID- 10679978 TI - Quantitative electrospray ionization mass spectrometric studies of ternary complexes of amino acids with Cu(2+) and phenanthroline. AB - Electrospray ionization (ESI) mass spectra of ternary complexes of Cu(2+) and 1,10-phenanthroline with the 20 essential amino acids (AA) were investigated quantitatively. Non-basic amino acids formed singly charged complexes of the [Cu(AA - H)phen](+) type. Lysine (Lys) and arginine (Arg) formed doubly charged complexes of the [Cu(HAA - H)phen](2+) type. Detection limits were determined for the complexes of phenylalanine (Phe), glutamic acid (Glu) and Arg, which were at low micromolar or submicromolar concentrations under routine conditions. Detection limits of low nanomolar concentrations are possible for amino acids with hydrophobic side-chains (Phe, Tyr, Trp, Leu, Ile) as determined for Phe. The efficiencies for the formation by ESI of gaseous [Cu(AA - H)phen](+) ions were determined and correlated with the acid-base properties of the amino acids, ternary complex stability constants and amino acid hydrophobicities expressed as the Bull-Breese indices (DeltaF). A weak correlation was found between DeltaF and the ESI efficiencies for the formation of gaseous [Cu(AA - H)phen](+) [Cu(HAA - H]phen](2+) and [AA + H](+) ions that showed that amino acids with hydrophobic side-chains were ionized more efficiently. In the ESI of binary and ternary amino acid mixtures, the formation of gas-phase Cu-phen complexes of amino acids with hydrophobic side-chains was enhanced in the presence of complexes of amino acids with polar or basic side-chains. An interesting enhancement of the ESI formation of [Cu(Glu - H)phen](+) was observed in mixtures. The effect is explained by ion cluster formation at the droplet interface that results in enhanced desorption of the glutamic acid complex. PMID- 10679979 TI - High-energy collision-induced dissociation of small polycyclic aromatic hydrocarbons. AB - High-energy collision-induced dissociation (CID) experiments on polycyclic aromatic hydrocarbons (PAHs) having 2-6 rings, naphthalene, anthracene, phenanthrene, fluoranthene, pyrene and coronene, were performed, and the relative abundances of their fragment ions were investigated as a function of collision energy. The results revealed that the PAHs except naphthalene showed a bimodal type distribution of positive fragmentation ions, which is closely similar to the fragment-ion distribution reported for the CID of three-dimensional fullerene, C(60)(+) and C(70)(+). The three-ring isomers of anthracene and phenanthrene and the four-ring isomers of fluoranthene and pyrene can be distinguishable in their spectra under an electron ionization energy of 70 eV, but the high-energy CID spectra of the three- and four-ring isomers were almost identical. The fragmentation corresponding to fragment ions in the low-mass region of the bimodal CID spectra could be interpreted by the simple statistical model that fragment ions are formed by random evaporation from the molecular ions after a considerable structural rearrangement, 'phase transition', occurring at some high energy state. PMID- 10679980 TI - Metastable decay of peptide ions on a Fourier transform mass spectrometer equipped with an external ion source. AB - Metastable decay rates of two peptides, RPPGFSPF and PKPQQFFGLM, were determined from ions produced in an external matrix-assisted laser desorption/ionization source with a Fourier transform mass spectrometer. An isolation and subtraction method that gives difference spectra was employed to monitor the product formation and metastable decays. The dependence of metastable decay rates on laser fluences and matrixes was demonstrated. PMID- 10679981 TI - Double transacetalization of diacylium ions. AB - A novel gas-phase reaction of diacylium ions of the O=C=X(+)=C=O type (X = N, CH) is reported: double transacetalization with cyclic acetals or ketals. The reaction is exothermic and highly efficient, and forms members of a new class of highly charged-delocalized ions: cyclic ionic diketals. Pentaquadrupole double- and triple-stage mass spectrometric (MS(2) and MS(3)) experiments reveal the high double transacetalization reactivity of O=C=N(+)=C=O and O=C=CH(+)=C=O, whereas the synthesis of differently substituted cyclic ionic diketals is performed in MS(3) experiments via sequential mono- and double transacetalization of O=C=N(+)=C=O and O=C=CH(+)=C=O with different acetals. With cyclic acetals, the acylium-thioacylium ion O=C=N(+)=C=S reacts promptly and selectively by mono transacetalization at its acylium site, but the free thiacylium site of its cyclic ionic ketal is nearly unreactive by double transacetalization. Therefore, only the acylium site of O=C=N(+)=C=S can be efficiently protected by transacetalization. Low-energy MS(3) collision-induced dissociation of the cyclic ionic diketals of O=C=N(+)=C=O and O=C=CH(+)=C=O sequentially frees each of the protected acylium site to form the mono-derivatized ion, and then the fully deprotected diacylium ion. PMID- 10679982 TI - Atmospheric pressure chemical ionization multi-stage mass spectrometry in the characterization of stereoisomeric synthons of cyclopropane amino acids. AB - The mass spectrometric behaviour of (1S,2R)-, (1R,2R)-, (1R,2S)- and (1S,2S)-2 [(S)-2,2-dimethyl-1, 3-dioxolan-4-yl]-1-spiro-?4'[2'-phenyl-5'(4'H)-oxazolone]? cyclopropane (2) and (1S,2R)-, (1R,2R)-, (1R,2S)- and (1S, 2S)-methyl-1-benzamido 2-[(S)-2,2-dimethyl-1, 3-dioxolan-4-yl]cyclopropanecarboxylate (3) was studied under atmospheric pressure ionization conditions and by multi-stage mass spectrometric (MS(n)) experiments performed with an ion trap. Interestingly, by using methanol as solvent, compounds 2 lead to [M + H + CH(3)OH](+) ions which, as proved by collisional experiments, exhibit the same structure of the corresponding compound 3. MS/MS of [MH](+) ions allows a clear characterization of the different stereoisomers, which give rise to specific fragmentation pathways, rationalized with respect to the structure of the neutral molecules. PMID- 10679983 TI - Determination of the carbohydrate composition and the disulfide bond linkages of bovine lactoperoxidase by mass spectrometry. AB - The extent and distribution of N-glycosylation and the nature of most of the disulfide bond linkages were determined for bovine lactoperoxidase through proteolytic and glycolytic digestions combined with matrix-assisted laser desorption/ionization mass spectrometric analysis. In addition, 98% of the primary sequence of the protein was confirmed. All five of the asparagines present in sequons were found to be glycosylated, predominantly by high mannose and complex structures. Six disulfide bonds were assigned, including Cys 32-Cys 45, Cys 146-Cys 156, Cys 150-Cys 174, Cys 254-Cys 265, Cys 473-Cys 530 and Cys 571-Cys 596. PMID- 10679984 TI - Determination of [(13)C]galactose enrichment in human plasma by gas chromatography/positive chemical ionization tandem mass spectrometry. AB - Galactosemia is a potentially fatal disease resulting from a deficiency of galactose-1-phosphate uridyl transferase. In order to perform mechanistic studies designed to elucidate further the etiology of the disease, we required a method to monitor (13)C enrichment in plasma galactose following a single oral dose or intravenous infusion of [1-(13)C]galactose. Determinations of plasma [(13)C]galactose enrichment requires methodology with extremely high specificity because of potential interference from other low molecular mass plasma constituents and from glucose, an isomer which is present in much higher concentrations. We have developed a method based on gas chromatography/positive chemical ionization tandem mass spectrometry (GC/PCI-MS/MS) for the precise and accurate determination of plasma [(13)C]galactose enrichment. The method employed a pentaacetylaldononitrile derivative of galactose in order to improve its GC and MS characteristics. Peak areas resulting from the transitions m/z 328 --> 106 and m/z 329 --> 107 were used to quantify the relative abundance of labeled and unlabeled galactose. Validation of the method was performed by determination of the precision and accuracy over a wide range of galactose concentrations and (13)C enrichments. The GC/PCI-MS/MS method was able to determine accurately enrichments at galactose concentrations down to 0.8 microM in the presence of 4 mM glucose, making it both highly selective and the most sensitive method currently available. PMID- 10679986 TI - Antisense peptide interactions studied by electrospray ionization mass spectrometry. AB - Non-covalent interactions between met- and leu-enkephalins and their antisense peptides were studied by electrospray ionization mass spectrometry. Mixtures of sense and antisense peptides gave both the corresponding homodimers and heterodimers. The relative abundance ratios of the heterodimer to that of the homodimer of the sense peptide and the relative stability constants of the heterodimers were compared with the corresponding values from mixtures of the sense peptides and a control peptide. The results show that there is a preferential interaction between the sense and antisense peptides compared with that between the sense and control peptides. PMID- 10679985 TI - Analysis of oxidized glycerophosphocholine lipids using electrospray ionization mass spectrometry and microderivatization techniques. AB - Oxidized low-density lipoprotein (LDL) is thought to play an important role in atherogenesis and cardiovascular disease in humans. Oxidized LDL is a complex mixture of many oxidized species, including numerous oxidized glycerophospholipids. Electrospray ionization and tandem mass spectrometry as well as microchemical derivatization of high-performance liquid chromatographically purified fractions derived from oxidized LDL were investigated as means to determine the structure of individual components present in oxidized LDL. One major oxidized phosphocholine lipid had an [M + H](+) ion at m/z 650. Derivatization to the trimethylsilyl ether and methoxime caused shifts in mass which, along with negative ion collision-induced dissociation mass spectra, were consistent with the presence of three species, 1-palmitoyl-2-(9 oxononanoyl)glycerophosphocholine and two isomeric 1-octadecanoyl-2 (hydroxyheptenoyl)glycerophosphocholines. These species were chemically synthesized. Trimethylsilylation of free hydroxyl groups increased the mass of the phospholipid acyl chains containing hydroxyl groups by 72 u. Conversion of carbonyl groups to the methoxylamine derivative increased the mass by 29 u. Ozonolysis of those products which contained double bonds proved to be a facile technique to determine the position and number of double bonds present. The use of these techniques was illustrated in the structural characterization of one major component (m/z 650, positive ions) in oxidized LDL as 1-octadecanoyl-2-(7 hydroxyhepta-5-enoyl)glycerophosphocholi ne. A possible mechanism for the formation of this unique chain-shortened glycerophospholipid is proposed. PMID- 10679987 TI - Tandem mass spectrometric analysis of (13)C-containing ions from a mixture of homologous peptides differing by one mass unit at a residue. AB - Tandem mass spectrometry of a mixture of two peptides that differ from each other by a single mass unit due to mutation is presented. The mutant beta-globin of hemoglobin Hoshida is present along with the normal counterpart, and the amino acid substitution of glutamine for glutamic acid is located within tryptic peptide T5 of M(r) 2057. 9. The mass of the mutated peptide is 1 u lower. In the isotopic cluster for the doubly charged ion of the peptide T5, the resolved ion with mass of 1030.0 represents the normal peptide with 93 (12)C atoms and the mutated one with 92 (12)C and one (13)C atoms. Collision-induced dissociation (CID) of this composite ion identified the mutation by presenting a key fragment derived from the (12)C-only mutant peptide, as reported in a previous study. Similarly, when an ion containing multiple (13)C atoms was selected as a precursor for CID, the mutation could be identified, even in large fragments, by a marked change in the shape of the isotopic cluster for the consecutive product ions. This study demonstrates the merit of selecting a resolved ion rather than the whole isotopic cluster as a precursor in the CID measurements of large peptides or proteins for characterizing heterozygous mutations. PMID- 10679988 TI - First generation and characterization of the enol of glycine, H(2)N--CH=C(OH)(2), in the gas phase. AB - The enol of glycine, H(2)N-CH&dbond;C(OH)(2), is generated in the gas phase by neutralization of the corresponding radical cation, which is available by dissociative electron ionization of isoleucine. Reionization approximately 0.6 micros later shows that the isolated enol (2) exists and does not isomerize to the significantly more stable glycine molecule, H(2)N--CH(2)--COOH (1); hence the intramolecular tautomerization 2-->1 must be associated with high barriers. The neutralization-reionization reactivity of 1(+*) further confirms that neutral glycine has a canonical structure (1) and is not a zwitterion. The unimolecular chemistry of 1(+*) is dominated by C--C bond cleavage to the immonium ion (+)H(2)NCH(2); in sharp contrast, 2(+*) primarily loses H(2)O. The ylide ion (+)H(3)N--CH(*)--COOH, an intermediate in the water loss from 2(+*), is found to readily equilibrate to 2(+*) prior to dissociation. Tautomers 1(+*) and 2(+*) differ in their charge-stripping behavior, with only 2(+*) forming a stable dication. The radical anions 1(-*) and 2(-*), formed by charge reversal of 1(+*) and 2(+*), respectively, dissociate extensively to (mainly) different closed shell fragment anions. An important channel is H(*) loss; 1(-*) yields the carboxylate ion H(2)N--CH(2)--COO(-) whereas 2(-*) yields the enolate ion H(2)N- CH=C(OH)O(-). PMID- 10679989 TI - Mapping of surrogate markers of cellular components and structures using laser desorption/ionization mass spectrometry. AB - Laser desorption/ionization mass spectrometry (LDI-MS) has been used to assess the potential of using surrogate markers, bound to cellular structures containing nucleic acids, to image or map the position of these structures within biological samples. In this study, organic dyes were used as markers because of their established use in the histochemical marking of nucleic acids, and also because they are amenable to LDI-MS. Eight cationic dyes were tested and all could be desorbed from nucleic acid samples without additional matrix after specifically binding to these molecules. Methylene Blue was the best of these based on its sensitivity to detection by LDI-MS and the fact that it can be washed from the tissue in areas where it was not specifically bound to provide low-intensity background signals. Experiments are reported which characterize the M(+) ion signal obtained from Methylene Blue with regard to sensitivity, reproducibility and possible use for quantitation. This dye was used to map (with a lateral resolution of 25 microm) several nucleic acid-containing samples spotted on prepared surfaces, and to image the location of nucleic acids in two model tissues, retinal vertical sections and thyroid whole mount sections. PMID- 10679990 TI - Location of alkali metal binding sites in endothelin A selective receptor antagonists, cyclo(D-Trp-D-Asp-Pro-D-Val-Leu) and cyclo(D-Trp-D-Asp-Pro-D-Ile Leu), from multistep collisionally activated decompositions. AB - We previously showed by using mass spectrometry that endothelin A selective receptor antagonists BQ123 and JKC301 form novel coordination compounds with sodium ions. This property may underlie the ability of an ET(A) antagonist to induce net tubular sodium reabsorption in the proximal tubule cells and reverse acute renal failure induced by severe ischemia. We have now defined the metal binding sites on BQ123 and JKC301 by subjecting the metal-containing peptides to multiple stages of collisionally activated decomposition (CAD) in an ion trap mass spectrometer. When submitted to low-energy CAD, the ring opens at the Asp Pro amide bond. The metal ion, which bonds, inter alia, to the carbonyl oxygen of the proline residue, acts as a fixed charge site, and directs a charge-remote, sequence-specific fragmentation of the ring-opened peptide. Amino acid residues are sequentially cleaved from the C-terminal end, and the terminal aziridinone structure moves one step toward the N-terminus with each C-terminal amino acid residue removed. These observations are the basis of a new method to sequence cyclic peptides. Amino acid residues are observed as sets of three ions, a*(n)PD, b*(n)PD and c*(n)PD where n is the number of amino acid residues in the peptide. PMID- 10679991 TI - Compassionate conservatism in urinary cytology. PMID- 10679992 TI - Fine-needle aspiration cytology of salivary gland: a review of 341 cases. AB - Three hundred and forty-one salivary gland fine-needle aspiration (FNA) cytology specimens taken over a 6-yr period were reviewed and correlated with clinical and/or histological findings. The aspirates were derived from parotid gland (212 cases), submandibular gland (124 cases), and minor salivary gland (5 cases). The major diagnostic categories were unsatisfactory (10 cases), normal (100 cases), sialadenitis (74 cases), cyst (34 cases), lipoma (5 cases), pleomorphic adenoma (55 cases), Warthin's tumor (36 cases), and malignancy (27 cases). The latter included 14 primary salivary neoplasms (4 lymphomas of mucosa-associated lymphoid tissue (MALT) type, 3 adenocarcinomas, 2 squamous carcinomas, 2 adenoid cystic cacinomas, and one case each of carcinoma ex pleomorphic adenoma, undifferentiated carcinoma, and high-grade mucoepidermoid carcinoma), and 13 metastases, 9 of which were derived from squamous carcinomas of head and neck origin. Clinicopathological review showed that 88 of 91 (97%) benign epithelial tumors and 27 of 31 (87%) malignant neoplasms with adequate FNA sampling were accurately diagnosed cytologically. False-negative results were caused by sampling error (7 cases), most notably in cystic tumors, or were due to misinterpretation of uncommon neoplasms (3 cases). The overall sensitivity, specificity, and accuracy were 92%, 100%, and 98%, respectively. FNA cytology provides accurate diagnosis of most salivary gland lesions and contributes to conservative management in many patients with nonneoplastic conditions. PMID- 10679993 TI - Cytology and flow cytometry of malignant effusions of multiple myeloma. AB - Identifying malignant plasma cells in body fluids from multiple myeloma patients is important for therapeutic and prognostic considerations. This can be difficult when plasma cells are mature in appearance or low in number. We examined the cytological and flow cytometric findings of myelomatous pleural and pericardial effusions from 8 patients with advanced multiple myeloma. Cytoplasmic immunoglobulin light chain excess vs. DNA ploidy in the plasma-cell population was evaluated by flow cytometry (FCM). The cytology smears of one pericardial and 14 pleural effusions from the 8 patients were reviewed. Screening Papanicolaou stained smears facilitated the detection of malignant nuclear features; however, morphology of plasma cells was best seen in Diff-Quik-stained smears. Low cellularity and inadequate air-drying of smears accounted for the false-negative cytology seen in two fluids from a single patient. A malignant plasma cell population was identified in 9 of 10 fluids submitted for FCM, including the two fluids with negative cytology. The false-negative FCM was from a suboptimal specimen with high background staining. Six fluids had an aneuploid DNA content, and four were diploid. A combination of Papanicolaou- and Diff-Quik-stained smears is recommended for the evaluation of plasma cells in effusions from patients with multiple myeloma. Cytology and flow cytometry confirmed malignancy in 87% and 90% of fluids evaluated, respectively; all cases were diagnosed by either one or both methods. Our results suggest that FCM and cytology of serous effusions in multiple myeloma patients are complementary and should be used in difficult cases. Diagn. Cytopathol. 2000;22:147-151. PMID- 10679994 TI - DNA flow cytometry of non-Hodgkin's lymphomas: correlation with cytologic grade and clinical relapse. AB - In this prospective study, we correlated cytological grading and clinical follow up of non-Hodgkin's lymphomas (NHL) with DNA flow cytometry (FCM) data from fine needle aspiration biopsy (FNAB) material. FNAB was performed from 34 successive cases of NHL, and the aspirated material was analyzed by DNA FCM. Cytological subtyping and grading were done by modified Working Formulation. Cases were followed up for 2.5 yr, and cytological grading, clinical follow-up, and DNA FCM data were correlated. There were 8 cases of low, 14 intermediate, and 12 high grade NHL. None of the cases of low-grade NHL showed DNA aneuploidy. Of the 8 DNA aneuploid cases, 5 were of intermediate and 3 were of high-grade NHL. Mean growth fraction (GF) of low-grade, intermediate-grade, and high-grade NHL was 6.1, 9.4, and 19.4, respectively. DNA aneuploidy was statistically significant (P < 0.05) between low- vs. intermediate- and high-grade NHL. Growth fraction was also statistically significant between low- vs. high-grade NHL. Six cases showed clinical recurrence, and one case died within 6 mo of diagnosis, due to widespread involvement of NHL. DNA aneuploidy and mean GF of recurrent and nonrecurrent cases were statistically significant (P < 0.05) irrespective of grading of NHL. DNA aneuploidy and GF are not well-correlated with morphological grading (by the Working Formulation). However, these two criteria are important for assessment of early clinical relapse of NHL. PMID- 10679995 TI - Incidental sampling of branchial remnants: a potential source of error in fine needle aspiration of neck lesions-a case report. AB - Remnants of the branchial apparatus can produce lesions in the head and neck region in later life, often amenable to fine-needle aspiration (FNA) diagnosis. Yet such remnants or rudimentary lesions can remain clinically undetected and can later interfere with the cytologic interpretation of other deep lesions of the neck, as the present case demonstrates. In this case the lesion, which by a subsequent resection turned out to be a neurilemmoma, had been adequately sampled by the FNA, yet the cytologic diagnosis was sidetracked by the presence in the specimen of immature squamous epithelial tissue fragments and other elements (multinucleated histiocytes, calcifications), on the basis of which the diagnosis of an epithelial lesion, likely malignant, was made. The neck surgery and a preceding endoscopic examination of the mouth, pharynx, and larynx did not identify such a lesion, but a detailed microscopic examination of the fibroadipose tissue between the tumor and the peripharyngeal region revealed the presence of epithelial microfragments with morphology partly corresponding to that of the FNA cytology, highly indicative of a branchiogenic lesion in the peripharyngeal region. The basic embryology of the branchial apparatus resulting in such defects is presented, as well as tentative guidelines for recognizing material deriving from accidental sampling of such lesions during FNA investigations of deep-seated masses of the neck. Diagn. Cytopathol. 2000;22:157 160. PMID- 10679996 TI - Peripheral primitive neuroectodermal tumor of the parotid gland region: report of a case with fine-needle aspiration findings. AB - A case of peripheral primitive neuroectodermal tumor of the parotid gland region in a 38-yr-old woman is reported. She had a 1-yr history of a large, firm, and slightly tender left parotid-region mass. CT scan showed an invasive tumor involving the parotid gland, mandible, infratemporal fossa, and parapharyngeal space. Fine-needle aspiration cytology of the mass showed a highly cellular, poorly cohesive smear pattern exhibiting small cuboidal cells, with fibrillary cytoplasm forming occasional rosette-like structures. Numerous intact single cells with fragile cytoplasm, finely granular chromatin, and inconspicuous nucleoli were present together with free-lying nuclei in the background. Histologic, immunohistochemical, and ultrastructural findings confirmed the diagnosis. Diagn. Cytopathol. 2000;22:161-166. Published 2000 Wiley-Liss, Inc. PMID- 10679997 TI - Nontyrosine crystalloids in salivary gland lesions: report of seven cases with fine-needle aspiration cytology and follow-up surgical pathology. AB - We report a series of seven patients who underwent fine-needle aspiration (FNA) for clinically apparent parotid gland lesions. In all seven cases, numerous to abundant polyhedral, multifaceted (nontyrosine) crystalloids were noted in the background of scanty cellular specimens composed predominantly of oncocytic cells. Subsequent surgical excision showed that three of the seven glands revealed sialolithiasis and sialadenitis without evidence of neoplasia. The histology of the remaining four cases consisted of two Warthin's tumor, one oncocytic papillary cystadenoma, and one cellular benign mixed tumor. In all seven cases the nontyrosine crystalloids were found in highest concentrations in cystic spaces lined with oncocytic metaplastic cells. We conclude that nontyrosine cystalloids can be associated with both neoplastic and nonneoplastic salivary gland disease, and they may be a product of oncocytic cell secretion. Diagn. Cytopathol. 2000;22:167-171. PMID- 10679998 TI - Fine-needle aspiration biopsy of postradiation epithelioid angiosarcoma of breast. AB - Angiosarcoma of breast skin and parenchyma is a rarely reported complication of irradiation for breast carcinoma. We report a case of a subareolar epithelioid angiosarcoma arising 8 years subsequent to lumpectomy and irradiation of the ipsilateral breast for infiltrating carcinoma. The epithelioid appearance of the neoplastic cells on fine-needle aspiration biopsy (FNA) biopsy suggested a recurrence of the primary carcinoma. Careful attention to certain cytomorphologic features and cell block immunohistochemistry were useful in the distinction from recurrent carcinoma. Cytologic features that identified this neoplasm as an angiosarcoma included marked cell discohesiveness, elongate cytoplasmic processes or "pseudopodia," heterogeneous cell size, large nucleoli or macronucleoli, and cytoplasmic lumina. Immunohistochemical markers, including Factor VIII antigen, CD31, and CD34, were positive, confirming the vascular nature of the neoplasm. Other markers ruled out morphologically similar neoplasms such as recurrent carcinoma and melanoma. Epithelioid angiosarcoma should be included in the differential diagnosis of a suspected recurrence of breast carcinoma several years postirradiation therapy. Diagn. Cytopathol. 2000;22:172-175. PMID- 10679999 TI - Cytology of polypoid adenomyomas: a report of two cases. AB - Uterine polypoid adenomyomas, both typical and atypical variants, often arise in the lower uterine segment or endocervical canal as pedunculated polypoid masses that may be accessible for cytologic sampling. However, their cytologic findings have rarely been described in the literature. Two women in their reproductive age presented with abnormal vaginal bleeding. The cervicovaginal smear of the first patient contained sheets and strips of reactive endocervical cells in an inflammatory background. In addition, loose aggregates of spindle-shaped smooth muscle cells were also noted. The findings were consistent with those of a typical polypoid adenomyoma. The cervicovaginal smears of the second patient consisted of tightly packed, crowded clusters of glandular cells which were initially interpreted as atypical glandular cells, suspicious of adenocarcinoma. In retrospect, loose aggregates of smooth muscle stromal cells were noted. Subsequent curettage revealed an atypical polypoid adenomyoma. The cytologic findings of typical polypoid adenomyoma were nonspecific except for the presence of loose aggregates of smooth muscle cells. The cytologic features of an atypical polypoid adenomyoma may mimic that of a neoplastic glandular process. The findings of tightly packed clusters of glandular cells and loose aggregate of bland-appearing smooth muscle cells in premenopausal patients may suggest the diagnosis of atypical polypoid adenomyoma. Diagn. Cytopathol. 2000;22:176-180. PMID- 10680000 TI - Cytology of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease). AB - Sinus histiocytosis with massive lymphadenopathy (SHML) is a benign, self limiting condition of unknown etiology, which generally presents as massive bilateral cervical lymphadenopathy. It is important to distinguish SHML from other causes of histiocytosis because of the different treatment modalities. This study was carried out to assess the utility of fine-needle aspiration cytology (FNAC) findings in SHML and to distinguish if from other reactive lymphadenopathies. The lymph nodes in 4 patients (3 male and 1 female) presenting with massive bilateral cervical lymphadenopathy were aspirated. All presented with persistent bilateral cervical lymphadenopathy, polymorphnuclear leukocytosis, and raised erythrocyte sedimentation rate (ESR). Smears showed a reactive lymphoid population consisting of mature lymphocytes, plasma cells, a few polymorphs, and many histiocytes showing emperipolesis. Based on the cytologic and clinical findings, a diagnosis of SHML was made. Histopathology confirmed the diagnosis in all cases. A conclusive diagnosis of SHML can be based on cytology, provided that the cytologic findings are interpreted in the appropriate clinical context. Biopsy can be avoided in these patients. Diagn. Cytopathol. 2000;22:181-185. PMID- 10680001 TI - Performance characteristics of the Indiana University Medical Center endometrial sampler (Tao Brush) in an outpatient office setting, first year's outcomes: recognizing histological patterns in cytology preparations of endometrial brushings. AB - Following successful hysterectomy-controlled trials, Tao brush endometrial cytology with liquid fixation was offered to clinicians for office use. This report recounts our first year's cytology and correlative histology outcomes. One hundred thirteen cases were accrued. Correlative tissue examinations comprising Pipelle (Prodimed, Neuilly-en-Thelle, France) biopsy, hysteroscopy and biopsy, dilatation and curettage, and hysterectomy were available at this institution for 59 cases. In 42 cases, cytology diagnoses could be compared to histology diagnoses. Twenty-five of 63 normal brushings were followed up. Fourteen were normal. Eleven Pipelle biopsies of cytologically atrophic endometrium were quantitatively limited and insufficient for diagnosis. Thirty-seven cases were abnormal, and 15 of these showed nuclear anaplasia. Twenty-eight of the abnormal cases were followed up. All correlative tissue examinations confirmed an abnormality. All 15 cases with nuclear anaplasia showed significant histopathology comprising atypical endometrial hyperplasia, endometrial intraepithelial neoplasia (EIN), endometrial intraepithelial carcinoma (EIC), and invasive adenocarcinoma. There were 13 inadequate endometrial brushings. Three cases had insufficient cellular material. The remaining 10 cases were cellular but were chiefly cervical/endocervical samples. Two of the cellular cases resulted from clinicians failing to replace the protective sheath over the brush bristles before removing the Tao brush from the endometrial cavity. The remaining 11 cases resulted from inaccessibility of the uterine cavity due to a tight or stenotic cervix. The Tao brush is a reliable uterine sampling device that performs well as a diagnostic tool for outpatient assessment of the endometrium of women with patent cervices. An advantage of endometrial cytology is that it accurately represents atrophic endometrium, and it is an effective case-finding tool for EIN and EIC. Women with tight or stenotic cervices are poor candidates for endometrial brushing, and may experience pain if the procedure is attempted. Diagn. Cytopathol. 2000;22:186-195. PMID- 10680002 TI - Expression of cytokeratin 19 in cytologic specimens of thyroid. PMID- 10680003 TI - Fine-needle aspiration of intra- and extraocular masses. PMID- 10680004 TI - Regarding the validity of cytology proficiency testing. PMID- 10680005 TI - Reply to dr. cramer PMID- 10680007 TI - Publisher's announcement PMID- 10680006 TI - Cytopathology in the United Kingdom: 1854 to the present. AB - The paper traces the history of cytopathology in the U.K. from the time of the pioneers in the last century to the 1930s and continues with the development of cervical and breast screening, with reference to training and quality control, to the present time. Diagn. Cytopathol. 2000;22:203-206. PMID- 10680008 TI - Cancer gene therapy: hard lessons and new courses. AB - Gene therapy for the treatment of cancer was initiated with high levels of optimism and enthusiasm. Recently, this perception has had to be tempered by the realisation that efficiency and accuracy of gene delivery remain the most significant barriers to its success. So far, there has been a disappointing inability to reach target cells with sufficient efficacy to generate high enough levels of direct killing and this has necessitated the invocation of bystander effects in order for any potential strategy to be convincing. At least in the foreseeable future, clinical advance will come from co-operation with other more established disciplines - such as chemotherapy, radiotherapy and immunotherapy. This is inevitable - and necessary - in order to prove that gene therapy can have efficacy as part of a combinatorial therapy, before hoping to move clinical mountains alone. In addition, there will have to be a thorough understanding of the clinical situations in which gene therapy will be used in order both to understand its own limitations, and to exploit its full potential. This will enable it to find the appropriate clinical niche in which its abilities will be optimally useful. Finally, anyone wishing to practise clinical cancer gene therapy will rapidly have to learn the ways of the free market and be able to juggle commercial necessities with ideological purity. Gene Therapy (2000) 7, 2 8. PMID- 10680009 TI - The application of gene therapy in autoimmune diseases. AB - The application of gene therapy in autoimmune disease represents a novel use of this technology. The goal of gene therapy in the treatment of autoimmune disease is to restore 'immune homeostasis' by countering the pro-inflammatory effects of the CD4+ T cells in the lesions of autoimmunity. This can be accomplished by adoptive therapy with transduced T cells which can specifically home to the site of inflammation and secrete 'regulatory' protein(s) to ameliorate the inflammation or by direct targeting of the retroviral vector to activated T cells in the sites of inflammation. Transduction of autoantigen recognizing CD4+ T cells, to secrete anti-inflammatory products, may become the 'magic bullet' to combat the ravages of autoimmune inflammation and tissue destruction. Gene Therapy (2000) 7, 9-13. PMID- 10680010 TI - Gene therapy in transplantation in the year 2000: moving towards clinical applications? AB - Transplantation faces several major obstacles that could be overcome by expression of immunomodulatory proteins through application of gene therapy techniques. Gene therapy strategies to prolong graft survival involve gene transfer of immunosuppressive or graft-protecting molecules. Very promising results have been obtained in small animal experimental models with inhibitors of co-stimulatory signals on T cells, immunosuppressive cytokines, donor major histocompatibility antigens and regulators of cell apoptosis or oxidative stress. The application of gene therapy techniques to transplantation offers a great experimental and therapeutic potential. Local production of immunosuppressive molecules may increase their therapeutic efficiency and reduce their systemic effects. When compared with other clinical situations, gene therapy in transplantation offers several potential advantages. Gene transfer into the graft can be performed ex vivo, during the transit between the donor and the recipient, thus avoiding many of the hurdles encountered with in vivo gene transfer. Furthermore, the difficulties associated with immune responses to the gene transfer vectors and transient gene expression may be easier to overcome when gene therapy protocols are applied to transplantation than when applied to other clinical situations. The next century should witness a rapid increase in the application of gene therapy techniques to large animal pre-clinical models of transplantation and later to clinical trials. Gene Therapy (2000) 7, 14-19. PMID- 10680011 TI - Lentiviral vectors: turning a deadly foe into a therapeutic agent. AB - The past 3 years have witnessed the spectacular irruption of lentiviral vectors into the limelight of the gene therapy scene. Owing to their ability to deliver transgenes in tissues that had long appeared irremediably refractory to stable genetic manipulation, lentivectors have opened fresh perspectives for the genetic treatment of a wide array of hereditary as well as acquired disorders, and a concrete proposal for their clinical use seems imminent. This article traces the path that has led to this rapid development and describes the current state of the art in the design and production of lentiviral vectors. The important question of biosafety is discussed. This system seems to have the edge over other gene delivery tools for particular targets, however, there remain several issues to be resolved before lentivectors make it to the bedside. Gene Therapy (2000) 7, 20-23. PMID- 10680012 TI - AAV vectors: is clinical success on the horizon? AB - Potential applications and impact of the adeno-associated virus (AAV) as a gene transfer vector have expanded rapidly in the last decade. Recent advances in the production of high-titer purified rAAV vector stocks have made the transition to human clinical trials a reality in the last moments of the millenium. Production improvements will be complemented in the coming years with understanding of and innovations in the targeting and packaging of rAAV, the design of transgene cassettes, and the host immune response to the vectors. These expected areas of progress are discussed, with special attention to clinical applications for which rAAV vectors may help close the gap towards successful gene therapy. Gene Therapy (2000) 7, 24-30. PMID- 10680013 TI - Nonviral gene therapy: promises and challenges. AB - The last 10 years have seen substantial progress in the development and application of nonviral vectors in gene therapy. However, many problems remain to be resolved before nonviral gene therapy can become a standard clinical practice. This review highlights the major breakthroughs in this field. The problems and future research directions are also discussed. Gene Therapy (2000) 7, 31-34. PMID- 10680014 TI - Functional balance between T cell chimeric receptor density and tumor associated antigen density: CTL mediated cytolysis and lymphokine production. AB - Genetically engineered expression of tumor-specific single chain antibody chimeric receptors (ch-Rec) on human T lymphocytes endow these cells with the parental monoclonal antibody (mAb) dictated tumor specificity and may be useful for clinical immuno-genetherapy. Therefore it was of importance to assess how the densities of tumor-specific receptors and tumor associated antigens (TAA), respectively, affect primary human T lymphocyte functions in relation to target cell susceptibilities to lysis. We therefore studied the functional balance between ch-Rec densities on human T lymphocytes and TAA on tumor cells. The gene construct encoding a ch-Rec derived from (1) a renal carcinoma cell (RCC) specific mouse mAb (G250), and (2) the human signal transducing Fc(epsilon)RI gamma-chain was used. To obtain ch-RecHIGH-POS and ch-RecLOW-POS T lymphocytes, two distinct retroviral vectors were used to introduce the gene constructs into primary human T lymphocytes. Levels of ch-Rec-redirected T lymphocyte mediated tumor cell lysis, as well as lymphokine production were determined using RCC lines as target/stimulator cells, which express either no or increasing densities of the TAA. A functional and dynamic balance between ch-Rec densities on cytotoxic T lymphocytes (CTLs) on the one hand and TAA densities on RCCs on the other, was found. In short, ch-RecHIGH-POS CTLs are triggered by TAAHIGH-POS as well as TAALOW-POS RCCs to lyse tumor cells and produce (IFN-gamma and TNF-alpha) lymphokine. In contrast, ch-RecLOW-POS T lymphocytes are only triggered for cytolysis and lymphokine production by relatively TAAHIGH-POS RCCs. In conclusion, (1) the activation of T lymphocyte responses is co-determined by the expression levels of the ch-Rec on T lymphocytes and the TAA on tumor cells and (2) at relatively high T lymphocyte ch-Rec expression levels the CTLs lyse tumor cells with a wide range of TAA densities. Gene Therapy (2000) 7, 35-42. PMID- 10680015 TI - Genetically modified CD34+ cells as cellular vehicles for gene delivery into areas of angiogenesis in a rhesus model. AB - To develop a cellular vehicle able to reach systemically disseminated areas of angiogenesis, we sought to exploit the natural tropism of circulating endothelial progenitor cells (EPCs). Primate CD34+ EPCs were genetically modified with high efficiency and minimal toxicity using a non-replicative herpes virus vector. These EPCs localized in a skin autograft model of angiogenesis in rhesus monkeys, and sustained the expression of a reporter gene for several weeks while circulating in the blood. In animals infused with autologous CD34+ EPCs transduced with a thymidine kinase-encoding herpes virus, skin autografts and subcutaneous Matrigel pellets impregnated with vascular growth factors underwent necrosis or accelerated regression after administration of ganciclovir. Importantly, the whole intervention was perfectly well tolerated. The accessibility, easy manipulation, lack of immunogenicity of the autologous CD34+ cell vehicles, and tropism for areas of angiogenesis render autologous CD34+ circulating endothelial progenitors as ideal candidates for exploration of their use as cellular vehicles when systemic gene delivery to those areas is required. Gene Therapy (2000) 7, 43-52. PMID- 10680016 TI - Effective suicide gene therapy in vivo by EBV-based plasmid vector coupled with polyamidoamine dendrimer. AB - This study demonstrates in vivo effectiveness of a nonviral vector system, Epstein-Barr virus (EBV)-based plasmid vector coupled with polyamidoamine (PAMAM) dendrimer (EBV/polyplex), in suicide gene therapy of cancer. The EBV-based vector is a plasmid vector containing EBV nuclear antigen 1 (EBNA1) gene and oriP from EBV genome. HSV-1 tk gene was transferred into Ewing's sarcoma cell lines, A4573 and KP-EWS-YI, by using an EBV-based plasmid vector, pSES.Tk, or a conventional plasmid vector, pS.Tk. Cells transfected with pSES.Tk/dendrimer showed approximately 100 times lower ID50 to ganciclovir (GCV) compared with those transfected with pS. Tk/dendrimer. Intratumoral injection of pSES.Tk/dendrimer but not pS. Tk/dendrimer drastically suppressed the growth of tumors which had generated from A4573 or Huh7 hepatocellular carcinoma (HCC) cells inoculated into severe combined immunodeficiency (SCID) mice. The treatment with pSES.Tk/dendrimer also resulted in significant prolongation of survival of the mice implanted with A4573. These results suggest that the EBV/polyplex system could be useful for in vivo suicide gene therapy of cancer. Gene Therapy (2000) 7, 53-60. PMID- 10680017 TI - Gene transfer facilitated by a cellular targeting molecule, reovirus protein sigma1. AB - To facilitate eventual genetic vaccination of mucosal tissues, a receptor mediated gene transfer system was devised using the reovirus adhesin, protein sigma1. Highly efficient uptake and internalization of protein sigma1 polylysine (PL) DNA complexes could be demonstrated by fluorescent microscopy. Successful cellular transfection of rodent and human cell lines was obtained with the recombinant protein sigma1 as a PL-DNA complex, and could be shown to be receptor specific. Transfection efficiency was dependent upon the ratio of DNA complexed to protein sigma1-PL and chloroquine treatment improved transfection efficiency dramatically. To test its ability to bind a mucosal inductive tissue, recombinant protein sigma1 was specifically bound to the nasal-associated lymphoid tissues (NALT). Thus, recombinant protein sigma1-PL-DNA conjugates can efficiently bind and transfect cells that express the receptor for protein sigma1. Gene Therapy (2000) 7, 61-69. PMID- 10680018 TI - Fas ligand gene-carrying adeno-5 AdEasy viruses can be efficiently propagated in apoptosis-sensitive human embryonic retinoblast 911 cells. AB - Recent reports have pointed out the difficulties in generating recombinant adenoviral vectors expressing FasL using eukaryotic cells. In the present study we cloned the murine FasL (mFasL) gene into the pCA14 or pShuttle vectors and recombined them with the adeno-5 virus backbone pBHG10 or pAdEasy-1 in eukaryotic (911 cells) or prokaryotic (E. coli) cells, respectively. Recombination of pCA14 mFasL with pBHG10 in Fas-carrying 911 cells leads to rapid expression of mFasL and cell death by apoptosis before virus replication is initiated. The same effect is observed when 911 cells are transfected with the pCA14-mFasL shuttle vector only. If recombination (pShuttle-mFasL with pAdEasy-1) is performed first in bacteria and 911 cells are transfected thereafter with recombined AdEasy mFasL, virus production starts immediately and the cells survive longer. The resulting AdEasy-mFasL viruses are able to infect other eukaryotic cells and induce expression of functional mFasL. This study describes a method for efficient generation of recombinant FasL adeno-5 viruses in eukaryotic cells. The method may be generally suitable for producing viruses carrying deleterious genes. Gene Therapy (2000) 7, 70-74. PMID- 10680020 TI - New tools for the generation of E1- and/or E3-substituted adenoviral vectors. AB - We have designed new vectors for the construction of recombinant adenoviruses containing expression cassettes in the E1 and/or E3 regions. Using a versatile set of restriction enzymes, the cassettes are cloned into small bacterial vectors and subsequently introduced into large plasmids containing the adenoviral sequences. Two positive selection markers facilitate the recovery of a cosmid containing a copy of the sequence of the recombinant adenovirus. The resulting cosmid is transfected into 293 or 911 cells in order to rescue the virus. Importantly, the method does not require any recombination event, either in E. coli or in mammalian cells. The entire procedure can generate viral plaques in 12 days. Gene Therapy (2000) 7, 80-87. PMID- 10680019 TI - A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax. AB - The bax gene plays a critical role in signaling apoptosis and expression through gene transfer may be valuable in the treatment of a variety of apoptosis-related diseases such as cancer. However, constructing an adenoviral vector expressing a bax gene driven by a constitutive promoter has been difficult, presumably because of the gene's high proapoptotic activity. Here we report a system that induces the expression of the bax gene safely by adenovirus-mediated gene cotransfer. Briefly, the system involves an adenoviral vector containing a human bax cDNA driven by a synthetic promoter consisting of five GAL4-binding sites and a TATA box (GT). This vector expresses a minimal background level of bax protein in cultured mammalian cells thus preventing apoptosis of packaging cells, however, expression of the bax gene can be induced substantially in vitro and in vivo by transferring it into target cells along with an adenoviral vector expressing the transactivator, fusion protein GAL4/VP16. Extensive apoptosis was observed after induction of the bax gene both in cultured human lung carcinoma cells and in the livers of Balb/c mice. Our results suggest that this GAL4 gene regulatory system provides an alternative approach to constructing viral vectors that express potentially toxic genes. Gene Therapy (2000) 7, 75-79. PMID- 10680021 TI - Guest Editor Dr B Hogan. PMID- 10680022 TI - Optimal therapeutic management of non-Q-wave myocardial infarction. PMID- 10680023 TI - Methods used to interpret the 12-lead electrocardiogram: Pattern memorization versus the use of vector concepts. AB - This article extols the value of using Grant's approach to the interpretation of electrocardiograms (ECGs). The essay includes a discussion on how people learn and emphasizes the difference in memorizing information, thinking, and learning. Simply stated, the brains of most people are not designed to memorize countless numbers of ECG patterns. Accordingly, the essay supports the view that a method of interpretation must be used, and the reader is encouraged to use basic principles of electrocardiography, including vector concepts, to interpret each ECG. PMID- 10680024 TI - Selective estrogen receptor modulators--a new age of estrogens in cardiovascular disease? AB - A large body of evidence suggests hormone replacement therapy (HRT) reduces cardiovascular risk in postmenopausal women. It is, however, associated with serious side effects, such as increased risk of breast and endometrial cancer. This has likely caused uneasiness among both women and health care providers. A new class of compounds, called selective estrogen receptor modulators (SERMs), have emerged. Through their interactions at the estrogen receptor level they have become a class of compounds distinct from estrogen. While they share similar effects with estrogen on such factors as lipid profile and bone density, they affect other tissues differently. Specifically, they do not induce endometrial hyperplasia and are therefore not associated with endometrial cancer. In vitro studies have also shown that they inhibit lipoprotein oxidation and vascular smooth muscle cell proliferation. The cumulative effects of these compounds may prove quite beneficial in reducing cardiovascular risk in postmenopausal women while avoiding serious side effects. This may, in turn, ease much of the anxiety surrounding the issue of HRT. Clinical trials are presently being conducted to evaluate the effectiveness of raloxifene, a SERM, on cardiovascular risk reduction in postmenopausal women. PMID- 10680025 TI - Anomalies in the dosing of diltiazem. AB - From early research, investigators understood that the dose of diltiazem required for the treatment of hypertension (commonly 360 mg/day) was greater than that required for the treatment of angina (commonly 240 mg/day). Nonetheless, studies of recent prescribing practices show that the 240 and 180 mg capsule strengths constitute more than 70% of the diltiazem prescriptions for hypertension. Physicians became accustomed to the lower antianginal doses of diltiazem for 7 years before a hypertension indication was approved. Subsequently, these dosing levels were reinforced by the production of once-a-day formulations with highest capsule strengths of 240 mg and 300 mg. These strengths were dictated by the sheer bulk of the formulations, which limited how much diltiazem could be inserted into the #00 capsule, the largest capsule that can be comfortably administered. An examination of the combined data from the six randomized, blinded, and placebo-controlled trials submitted to the FDA for the original new drug applications of the three formulations of diltiazem available in the United States shows a clear linear dose-response relationship between diltiazem dose and blood pressure lowering through the 480-540 mg/day range. It also demonstrates that the 90-120 mg/day range is the "no-effect dose." These conclusions are supported by a MEDLINE review of all other studies of multilevel dosing of higher dose levels of diltiazem. The data support the conclusion that diltiazem is generally underdosed, but when properly dosed may be the single most potent antihypertensive overall. PMID- 10680026 TI - Dipyridamole-induced reversible thallium-201 defect in patients with vasospastic angina and nearly normal coronary arteries. AB - BACKGROUND: Dipyridamole is a vasodilator of resistance vessels and has no vasoconstrictive effect on large coronary arteries. HYPOTHESIS: The present study used dipyridamole thallium-201 (201Tl) scintigraphy to clarify the role of microvasculature in coronary flow limitation in patients with vasospastic angina and normal coronary arteries. METHODS: Sixteen patients underwent dipyridamole and exercise 201Tl scintigraphy and provocative coronary angiography with acetylcholine. All patients showed coronary spasm (> 90% vasoconstriction concomitant with ST change) in at least one vessel. RESULTS: Dipyridamole or exercise caused only ST depression despite the ST elevation observed at spontaneous attack. Dipyridamole 201Tl scintigraphy demonstrated the reversible defects (11 cases), as did exercise 201Tl scintigraphy (13 cases). The region of 201Tl defect was not always consistent with the territories of angiographically depicted vasospastic arteries. Basal coronary tone, assessed by percentage of diameter change of epicardial arteries from baseline to isosorbide dinitrate administration, did not differ between the positive and the negative defect regions. CONCLUSION: These results support the hypothesis that, in addition to epicardial coronary spasm, the dysfunction of microvasculature is responsible for abnormal coronary perfusion in the subgroup of patients with vasospastic angina and normal coronary arteries. PMID- 10680027 TI - Rest and exercise hemodynamics before and after valve replacement--a combined Doppler/catheter study. AB - BACKGROUND: Hemodynamic improvement is a common finding following valve replacement. However, despite a normally functioning prosthesis and normal left ventricular ejection fraction, some patients may show an abnormal hemodynamic response to exercise. METHODS: In a combined catheter/Doppler study, rest and exercise hemodynamics were evaluated in 23 patients following aortic (n = 12) (Group 1) or mitral valve (n = 11) (Group 2) replacement and compared with preoperative findings. Patient selection was based on absence of coronary artery disease and left ventricular failure as shown by preoperative angiography. Cardiac output, pulmonary artery pressure, pulmonary capillary pressure, and pulmonary resistance were measured by right heart catheterization, whereas the gradient across the valve prosthesis was determined by Doppler echocardiography. Postoperative evaluation was done at rest and during exercise. The mean follow-up was 8.2 +/- 2.2 years in Group 1 and 4.2 +/- 1 years in Group 2. RESULTS: With exercise, there was a significant rise in cardiac output in both groups. In Group 1, mean pulmonary pressure/capillary pressure decreased from 24 +/- 9/18 +/- 9 mmHg preoperatively to 18 +/- 2/12 +/- 4 mmHg postoperatively (p < 0.05), and increased to 43 +/- 12/30 +/- 8 mmHg with exercise (p < 0.05). The corresponding values for Group 2 were 36 +/- 12/24 +/- 6 mmHg preoperatively, 24 +/- 7/17 +/- 6 mmHg postoperatively (p < 0.05), and 51 +/- 2/38 +/- 4 mmHg with exercise (p < 0.05). Pulmonary vascular resistance was 109 +/- 56 dyne.s.cm-5 preoperatively, 70 +/- 39 dyne.s.cm-5 postoperatively (p < 0.05), and 70 +/- 36 dyne.s.cm-5 with exercise in Group 1. The corresponding values for Group 2 were 241 +/- 155 dyne.s.cm-5, 116 +/- 39 dyne.s.cm-5 (p < 0.05), and 104 +/- 47 dyne.s.cm-5. There was a significant increase in the gradients across the valve prosthesis in both groups, showing a significant correlation between the gradient at rest and exercise. No correlation was found between valve prosthesis gradient and pulmonary pressures. CONCLUSION: Exercise-induced pulmonary hypertension and abnormal left ventricular filling pressures seem to be a frequent finding following aortic or mitral valve replacement. Both hemodynamic abnormalities seem not to be determined by obstruction to flow across the valve prosthesis and may be concealed, showing nearly normal values at rest but a pathologic response to physical stress. PMID- 10680028 TI - Lipid-altering efficacy and safety of simvastatin 80 mg/day: long-term experience in a large group of patients with hypercholesterolemia. World Wide Expanded Dose Simvastatin Study Group. AB - BACKGROUND: Elevated levels of low-density lipoprotein (LDL) cholesterol promote the development of atherosclerosis and coronary heart disease. HYPOTHESIS: Simvastatin 80 mg/day will be more effective than simvastatin 40 mg/day at reducing LDL cholesterol and will be well tolerated. METHODS: Two similar, randomized, multicenter, controlled, double-blind, parallel-group, 48-week studies were performed to evaluate the long-term lipid-altering efficacy and safety of simvastatin 80 mg/day in patients with hypercholesterolemia. One study conducted in the US enrolled patients meeting the National Cholesterol Education Program (NCEP) LDL cholesterol criteria for pharmacologic treatment. In the other multinational study, patients with LDL cholesterol levels > or = 4.2 mmol/l were enrolled. At 20 centers in the US and 19 countries world-wide, 1,105 hypercholesterolemic patients, while on a lipid-lowering diet, were randomly assigned at a ratio of 2:3 to receive simvastatin 40 mg (n = 436) or 80 mg (n = 669) once daily for 24 weeks. Those patients completing an initial 24-week base study were enrolled in a 24-week blinded extension. Patients who had started on the 80 mg dose in the base study continued on the same dose in the extension, while those who had started on the 40 mg dose were rerandomized at a 1:1 ratio to simvastatin 40 or 80 mg in the extension. RESULTS: There was a significant advantage in the LDL cholesterol-lowering effect of the 80 mg dose compared with that of the 40 mg dose, which was maintained over the 48 weeks of treatment. The mean percentage reductions (95% confidence intervals) from baseline in LDL cholesterol for the 40 and 80 mg groups were 41% (42, 39) and 47% (48, 46), respectively, for the 24-week base study, and 41% (43, 39) and 46% (47, 45), respectively, after 48 weeks of treatment (p < 0.001 between groups). Larger reductions in total cholesterol and triglycerides were also observed with the 80 mg dose compared with the 40 mg dose at Weeks 24 and 48. Both doses were well tolerated, with close to 95% of patients enrolled completing the entire 48 weeks of treatment. Myopathy (muscle symptoms plus creatine kinase increase > 10 fold upper limit of normal) and clinically significant hepatic transaminase increases (> 3 times the upper limit of normal) occurred infrequently with both doses. There was no significant difference between the groups in the number of patients with such increases, although there were more cases for both with the 80 mg dose. CONCLUSIONS: Compared with the 40 mg dose, simvastatin 80 mg produced greater reductions in LDL cholesterol, total cholesterol, and triglycerides. Both doses were well tolerated. PMID- 10680029 TI - Incidence and significance of profound hypotension during dobutamine stress echocardiography. AB - BACKGROUND: Mild hypotension (drops of systolic blood pressure of > or = 20 mmHg) occurs in 14-38% of dobutamine stress echo (DSE) and carries a good prognosis for subsequent cardiac events. The incidence and significance of more profound hypotension (PH) (> or = 50 mmHg) is unknown. HYPOTHESIS: The aim of the study was to determine the incidence of PH during DSE and its prognosis for subsequent cardiac events. METHODS: We reviewed 617 DSE performed at our institution between 1992 and 1996 and identified two DSE subgroups. The first group (PH group) consisted of all patients with PH during DSE. A second group was selected with baseline characteristics similar to the PH group but without PH during DSE (non PH group). Follow-up was by a physician chart review and direct telephone contact. Cardiac event rates were determined for hard [myocardial infarction (MI), or cardiac death] and soft (angina, congestive heart failure, coronary angioplasty, or coronary bypass surgery) events occurring after the DSE. RESULTS: Of the 617 DSE performed, 16 (3%) patients developed PH (PH group) during DSE, with 13 showing no inducible ischemia. The hard and soft cardiac event rate in this 13 PH group was 46% (mean follow-up of 28.7 +/- 18 months). Of the non-PH group, 32 patients had a negative DSE with a coronary event rate of 12.5%. Profound hypotension correlated with a significantly higher cardiac event rate (p < 0.02). CONCLUSIONS: The incidence of PH during DSE is low (3%) and appears to predict a worse prognosis for subsequent cardiac events. PMID- 10680030 TI - Tricuspid stenosis and regurgitation: Doppler and color flow echocardiography and cardiac catheterization findings. PMID- 10680031 TI - Cardiac hemangioma. PMID- 10680032 TI - Fungal endocarditis of the mitral valve. PMID- 10680033 TI - Tachycardia-dependent right bundle-branch block with supernormal conduction. AB - This paper reports the case of a 76-year-old man in whom atrial flutter with varying atrioventricular block and intermittent right bundle-branch block was found. This is the first report on tachycardia-dependent right bundle-branch block associated with supernormal conduction in a case of atrial flutter. When an impulse is conducted to the ventricles beyond 0.72 s after a QRS complex of right bundle-branch block configuration, the impulse falls after the abnormally long effective refractor period of the right bundle branch and passes through the right bundle branch. When the conducted impulse occurs within 0.72 s after a QRS complex of right bundle-branch block configuration, the impulse usually falls in the refractory period and is blocked in the right bundle branch; however, only when the impulse occurs 0.48 or 0.49 s after that does it fall in the supernormal period and passes through the right bundle branch. The findings in the present report strengthen our previous suggestion that the presence of supernormal conduction plays an important role in the initiation of reentrant ventricular tachycardia. PMID- 10680034 TI - Antidromic atrioventricular reentrant tachycardia mimicking ventricular tachycardia in the setting of previous myocardial infarction. AB - The differentiation between ventricular tachycardia and broad-complex supraventricular tachycardia can be extremely difficult, particularly in emergency situations. We report a case of hemodynamically compromising broad complex tachycardia in a 63-year-old man. The patient had previously sustained an anteroseptal myocardial infarction and had subsequently undergone coronary artery bypass surgery because of triple-vessel coronary artery disease. Intravenous treatment with ajmalin terminated the tachycardia and revealed preexcited QRS complexes compatible with the presence of a left-sided atrioventricular accessory pathway. An antidromic atrioventricular reentrant tachycardia (identical to the clinical tachycardia) was induced during an electrophysiologic study. In conclusion, there are several causes of broad-complex tachycardia, even in patients with previous myocardial infarction, and, where doubt exists, electrophysiologic studies should be performed. PMID- 10680035 TI - John B. Barlow: master clinician and compleat cardiologist. PMID- 10680036 TI - Introduction: Expanding the horizons in unstable coronary artery disease. PMID- 10680037 TI - Can we differentiate the low-molecular-weight heparins? AB - The low-molecular-weight heparins (LMWHs) have a number of therapeutic advantages, relative to standard unfractionated heparin (UFH). They are readily bioavailable when injected subcutaneously and can be given in fixed doses, allowing for far simpler administration. Several LMWHs are now commercially available, each demonstrating different physical and chemical properties and different activities in animal models of anticoagulation or hemorrhage. In clinical comparisons with placebo in the treatment of unstable coronary artery disease (UCAD), the LMWHs dalteparin sodium and nadroparin calcium have demonstrated good anticoagulant efficacy. In comparisons with UFH, on the other hand, only enoxaparin has shown superior anticoagulant activity, as reported in the results of the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-wave Coronary Events (ESSENCE) and Thrombolysis In Myocardial Infarction (TIMI) 11B trials. However, close scrutiny of the methodology of the clinical trials in UCAD reveals considerable differences in study designs, dosage regimens, duration of administration of active treatments, and the timing and definition of endpoints. Therefore, it would not be scientifically sound to compare results with the different LMWHs based on the current available studies. It is also not possible to draw any conclusions with regard to the relative efficacy of the different LMWHs, since there are no properly-sized comparative data between dalteparin sodium, enoxaparin sodium, and nadroparin calcium. PMID- 10680038 TI - Acute management--how should we intervene? AB - A crucial question in the acute management of the patient with unstable coronary artery disease (UCAD) is whether to carry out early intervention, performing angiography soon after presentation and following this with revascularization where appropriate, or whether to follow a noninvasive medical strategy as far as possible unless symptoms necessitate intervention. The body of literature addressing this question is sparse, but the recent Fast Revascularization during InStability in Coronary artery disease (FRISC II) study has provided new insights into the problem. Using a factorial design to randomize patients to invasive or noninvasive management strategies, and to short- or long-term treatment with the low-molecular-weight heparin (LMWH) dalteparin sodium (Fragmin), it was shown in FRISC II that early invasive treatment (within 7 days), when combined with optimal medical pretreatment with dalteparin sodium, aspirin, and appropriate antianginal medication, is associated with improved clinical outcomes, relative to a "watchful waiting" approach based on noninvasive therapy. Thus, an early invasive approach following aggressive medical pretreatment should be the preferred strategy for patients with UCAD who present with signs of ischemia on the electrocardiogram or raised biochemical markers of myocardial damage at admission. PMID- 10680039 TI - Long-term management--the way forward? AB - The mainstay of treatment for unstable coronary artery disease (UCAD) currently consists of antithrombotic therapy with aspirin plus unfractionated heparin (UFH), together with anti-ischemic treatment with beta blockers and nitrates. Recently, there has been a trend toward replacement of UFH with low-molecular weight heparins (LMWHs), since these products offer significant advantages over the parent compound. Several lines of evidence suggest that prolongation of treatment with LMWHs beyond the acute phase may be appropriate in patients with UCAD. The Fragmin and Fast Revascularization during InStability in Coronary artery disease (FRISC II) study was designed to evaluate this hypothesis using the LMWH dalteparin sodium (Fragmin). A factorial design was used to randomize patients enrolled in the FRISC II study to an invasive or noninvasive management strategy, and to treatment with dalteparin sodium or placebo. Treatment with dalteparin sodium significantly reduced incidences of death and/or myocardial infarction (MI) during the first months of treatment (the reduction in the relative risk of double endpoint events was statistically significant at 47.0% at 1 month, and remained so at 2 months, but was no longer statistically significant at the 3-month assessment). However, risk, as defined by the triple endpoint of death, MI, and revascularization, was significantly lower (13.0% relative risk reduction) at 3-month follow-up in the treatment group randomized to dalteparin sodium than among patients receiving placebo. In patients in whom revascularization procedures were carried out, the risk of new, postprocedural events was low in both the placebo and dalteparin sodium arms. Thus, dalteparin sodium appears to protect patients from cardiac events until they undergo invasive procedures, and it can therefore be used as a bridge to revascularization. PMID- 10680040 TI - Targeting treatment for optimal outcome. AB - Rapid assessment of patients presenting with acute chest pain is essential, in order to distinguish between those who have a life-threatening condition, such as myocardial infarction or unstable angina, and the substantial proportion who do not have an acute coronary syndrome. It is thus of vital importance that reliable techniques are available to facilitate rapid risk stratification, as an aid to both clinical diagnosis and management strategy decisions. Assessments based on clinical findings, electrocardiographic monitoring, symptom-limited exercise testing, and biochemical marker measurements, used either singly or in various combinations, can fulfill this role. The present paper reviews some of the recent data that demonstrate the value of these techniques. Very few studies allow conclusions to be drawn about optimal treatment strategies in relation to groups stratified according to prognostic markers, and the question of whether intense medical treatment or early invasive intervention is most beneficial is one that clinical trials have yet to address adequately. In the recently completed Fragmin and Fast Revascularization during InStability in Coronary artery disease (FRISC II) study, comparisons were made of clinical outcomes achieved with early invasive versus noninvasive (i.e., medical) management strategies, and with short term versus prolonged anticoagulation with dalteparin sodium (Fragmin), in patients with unstable coronary artery disease. All study participants underwent symptom-limited exercise testing and provided blood sample for measurements of biochemical markers; continuous electrocardiography monitoring and echocardiography were also performed in a high proportion of patients. Data from the FRISC II trial thus shed further light on the issue of risk stratification and its use to determine optimal treatment strategies. PMID- 10680041 TI - Regulation of macrophage gene expression by peroxisome-proliferator-activated receptor gamma: implications for cardiovascular disease. AB - The peroxisome-proliferator-activated receptor gamma is a member of the nuclear receptor superfamily that functions as a key transcriptional regulator of cell differentiation and lipid metabolism. In addition, peroxisome-proliferator activated receptor gamma is now recognized to be the biological receptor for the thiazolidinedione class of antidiabetic drugs, which includes troglitazone and rosiglitazone. Recent evidence indicates that peroxisome-proliferator-activated receptor gamma is expressed at high levels in macrophages, including the foam cells of atherosclerotic lesions. Oxidized low-density lipoprotein, which plays a central role in lesion development, can activate peroxisome-proliferator activated receptor gamma by providing the cell with oxidized fatty acid ligands of the receptor. The elucidation of a peroxisome-proliferator-activated receptor gamma signalling pathway in macrophages provides a mechanism by which oxidized lipids may directly regulate gene expression in the context of the atherosclerotic lesions. A number of potential target genes for peroxisome proliferator-activated receptor gamma in these cells have been identified. Some, such as the type B scavenger receptor CD36 are induced by peroxisome-proliferator activated receptor gamma ligands, whereas others, such as scavenger receptor type A, inducible nitric oxide synthetase and certain cytokines, are repressed. Given the widespread clinical use of thiazolidinediones, it is important to consider the influence of these drugs on the risk of atherosclerosis. The net effect of peroxisome-proliferator-activated receptor gamma ligands on the atherogenic process is likely to reflect a balance between local effects in the artery wall and systemic effects on lipid metabolism. PMID- 10680042 TI - Influence of the HDL receptor SR-BI on atherosclerosis. AB - The scavenger receptor class B, type I (SR-BI) is an HDL receptor that mediates selective cholesterol uptake from HDL to cells. In rodents, SR-BI has a critical influence on plasma HDL-cholesterol concentration and structure, the delivery of cholesterol to steroidogenic tissues, female fertility, and biliary cholesterol concentration. SR-BI can also serve as a receptor for non-HDL lipoproteins and appears to play an important role in reverse cholesterol transport. Recent studies involving the manipulation of SR-BI expression in mice, either using adenovirus-mediated or transgenic hepatic overexpression or using homologous recombination for complete functional ablation, indicate that the expression of SR-BI protects against atherosclerosis. If SR-BI has a similar activity in humans, it may become an attractive target for therapeutic intervention. PMID- 10680043 TI - Pathology of in-stent restenosis. AB - The process of in-stent restenosis parallels wound healing responses. Stent deployment results in early thrombus deposition and acute inflammation, granulation tissue development, and ultimately smooth muscle cell proliferation and extracellular matrix synthesis. The severity of arterial injury during stent placement correlates with increased inflammation and late neointimal growth. These pathological findings provide useful targets for therapies aimed at reducing the incidence of in-stent restenosis. PMID- 10680044 TI - Smooth muscle cell phenotypes in atherosclerotic lesions. AB - Recently, there has been a dramatic change in the way we think about the role of vascular smooth muscle cells in atherosclerosis, and it is now generally accepted that a dearth of vascular smooth muscle cells in an atherosclerotic plaque is a detrimental feature of the disease. Indeed, it is now recognized that the phenotypes of vascular smooth muscle cells within a plaque dictate its features, progression and stability. Therefore an understanding of the processes that generate and regulate vascular smooth muscle cell heterogeneity are of critical importance for future therapeutic advancement in the treatment of atherosclerosis. PMID- 10680045 TI - Functional genomics and DNA array techniques in atherosclerosis research. AB - DNA arrays are revolutionizing the analysis of gene expression and single nucleotide polymorphisms of genomic DNA. Currently, the expression of 10-15% of human genes can be analysed simultaneously in a single experiment using cDNA or oligonucleotide-based format of DNA array. Alternatively, smaller DNA arrays with a limited number of selected genes, such as cytokines, growth factors or transcription factors, can be used. In concordance with Human Genome Project, after a few years, the DNA arrays will allow the analysis of expression of the whole human genome and will have a great impact on basic research, drug development and diagnostics. It is important to characterise mechanisms of atherosclerosis-related diseases at the level of gene expression so that new therapeutic strategies can be identified. With the aid of DNA array it is possible to identify multiple, simultaneous, transcriptional events that ameliorate or contribute to atherogenesis. The results are non-physical maps of the function, hierarchy and interactions of genetic programs. In this review we focus on DNA array technology and its applications in atherosclerosis research. PMID- 10680046 TI - Clinical lipidology at the end of the millennium. PMID- 10680047 TI - Modification of coronary artery disease progression by cholesterol-lowering therapy: the angiographic studies. AB - Large randomized placebo-controlled trials have demonstrated that cholesterol lowering with statin therapy reduces the incidence of adverse cardiac events. Smaller angiographic studies have shown that coronary artery disease progression can be slowed and, in some cases, reversed by cholesterol-lowering interventions. These anatomical changes, however, are small and occur too slowly to account for the early clinical benefit. Current evidence suggests that plaque stabilization is the most important mechanism, by which cholesterol-lowering therapy reduces both the incidence of adverse cardiac events and coronary artery disease progression. PMID- 10680048 TI - Effects of statins on carotid disease and stroke. AB - The results of recent trials indicate that statin treatment reduces not only the risk of coronary heart disease, but also the risk of stroke, in patients with existing heart disease. The need for the treatment of such patients is now generally recognized. Mechanisms for risk reduction include the retardation of plaque progression, plaque stabilization, and reducing the risk of coronary events. Questions remain regarding the discrepancy between epidemiological data and statin trials data, the precise mechanism of action of statins, and their role in the prevention of recurrent stroke in individuals who have experienced a previous stroke or transient ischemic attack but are free of coronary disease. PMID- 10680049 TI - Low-density lipoprotein-independent effects of statins. AB - Statins have pleiotropic properties that complement their cholesterol-lowering effects. These properties may partly account for their established benefit in the prevention of coronary artery disease beyond the reduction of LDL-cholesterol levels. The most widely recognized properties are reviewed here. They include: (i) nitric oxide-mediated improvement of endothelial dysfunction and upregulation of endothelin-1 expression; (ii) antioxidant effects; (iii) anti-inflammatory properties; (iv) inhibition of cell proliferation with anticarcinogenic actions in animals; (v) stabilization of atherosclerotic plaques; (vi) anticoagulant effects; and (vii) inhibition of graft rejection after heart and kidney transplantation. As advances are made in our knowledge, new properties are steadily being uncovered. Pleiotropic effects are currently being given consideration when instituting combination therapy for patients at high cardiovascular risk. Some pleiotropic effects are negative, and may account for occasional untoward drug interactions. For many of these new properties, the clinical relevance has not been established. The challenge for the future will be to design and carry out appropriate clinical trials to establish their relative importance in the prevention of coronary artery disease. PMID- 10680050 TI - Fibrates, dyslipoproteinaemia and cardiovascular disease. AB - Recent epidemiological data have reaffirmed that elevated plasma triglyceride and low HDL-cholesterol levels are important risk factors for atherosclerotic vascular disease. The rationale for the clinical use of fibric acid derivatives, which are designed to correct this metabolic nexus, is now on firmer ground. The mechanism of action of fibrates on lipoprotein metabolism has recently been elucidated at the molecular level and involves the activation of peroxisome proliferator-activated receptor-alpha 1 in the liver, with the net effect of improving the plasma transport rates of several lipoproteins. Other potential anti-atherothrombotic effects include the inhibition of coagulation and enhancement of fibrinolysis, as well as the inhibition of inflammatory mediators involved in atherogenesis. These consequences probably underpin the favourable effects of fibrates seen in recent angiographic and clinical trials. Two important clinical trials on the effect of gemfibrozil (Veterans Administration HDL-Cholesterol Intervention Trial) and bezafibrate (Bezafibrate Infarction Prevention Study) have recently been completed in subjects with elevated triglyceride, low HDL and normal or near-normal LDL-cholesterol levels. The results testify to the efficacy of these agents in decreasing the incidence of cardiovascular events, particularly in patients with multiple risk factors and plasma triglyceride levels of over 2.2 mmol/l. The findings of these trials are compared with the statin-based Air Force/Texas Coronary Atherosclerosis Prevention Study, with a recommendation that future studies in appropriately selected patients should examine the synergistic effect of the fibrate/statin combination. The absolute risk reduction in the incidence of coronary events in the Veterans Administration-HDL-Cholesterol Intervention Trial compares favourably with the statin trials. The therapeutic aspects of the efficacy and safety of fibrates are reviewed. Besides primary mixed hyperlipidaemias, particular indications for the clinical use of fibrates include type 2 diabetes, the metabolic syndrome and renal insufficiency. The St Mary's, Ealing, Northwick Park Diabetes Cardiovascular Disease Prevention Study has suggested that fibrates may decrease the incidence of coronary events in type 2 diabetes, but this hypothesis will be more extensively tested in the Diabetes Atherosclerosis Intervention Study, Fenofibrate in Event Lowering in Diabetes Study and Lipids in Diabetes Study projects. Although significant new knowledge has accrued over the past few years concerning the fundamental and clinical aspects of fibrates, the success of these agents in clinical practice depends on the availability of methods for assessing cardiovascular risk as well as on treatment guidelines, which as presently designed and recommended may be inaccurate and suboptimal. PMID- 10680051 TI - Non-pharmacological lowering of low-density lipoprotein by apheresis and surgical techniques. AB - In the past year, new data have appeared on the long-term benefits of low-density lipoprotein apheresis in severely hypercholesterolemic patients who are refractory to lipid-lowering drug therapy. Such data are critical for clinical decision-making, because they confirm the hypothesis that the dramatic reduction in low-density lipoprotein made possible by this technique produces clear-cut clinical benefits. Because of its efficacy and low incidence of side-effects, apheresis for severe drug-refractory hypercholesterolemia has superseded surgical approaches, such as liver transplantation or ileal bypass. PMID- 10680052 TI - Hormone replacement therapy and cardiovascular disease. AB - This year's work on hormone replacement therapy (HRT) and cardiovascular disease has been remarkable for the publication of the first randomised controlled trial of HRT use, the Heart Estrogen Replacement Study (HERS). The findings go against not only the trend of previous observational epidemiological studies, but also against findings in the very many studies which have previously shown and continue to show this year a beneficial effect of HRT on a large variety of cardiovascular risk factors, including endothelial function, here reviewed. The aspect of the effect of HRT on clotting variables is clearly crucial given the increased risk of venous thrombosis, and also increased number of cardiac events in the first 4 months of the HERS. Prothrombotic factors increase with age in women, and HRT alters these, particularly fibrinogen, factor VII, and PAI (less change with transdermal HRT) and antithrombin III. In normal women therefore the balance should be towards fibrinolysis rather than coagulation. Work has been presented in abstract for clarifying the effects of HRT on coagulation markers and grasping the problem of differences according to its route of administration. The full publications on this work are expected shortly. We are still awaiting evidence from randomized controlled trials of HRT in primary prevention; one is now recruited but will not report until 2005. PMID- 10680053 TI - Dietary and pharmacological antioxidants in atherosclerosis. AB - Antioxidants that inhibit LDL oxidation are thought to be potential anti atherogenic compounds. The results of major human randomized trials with antioxidants have, however, been disappointing, except for probucol, which consistently inhibits restenosis. Similarly, animal intervention studies show that antioxidants do not generally inhibit atherosclerosis, although some compounds provide protection. Direct evidence for the oxidation of LDL causing atherosclerosis is needed. This article summarizes results from antioxidant intervention studies, and highlights some of the key issues that need to be addressed to link biochemical changes in the arterial wall more directly to the oxidation theory of atherosclerosis. PMID- 10680054 TI - Bibliography. Current world literature. Atherosclerosis: cell biology and lipoproteins. PMID- 10680055 TI - Bibliography. Current world literature. Therapy and clinical trials. PMID- 10680056 TI - Nutrition. PMID- 10680057 TI - Genetics and molecular biology. PMID- 10680058 TI - Lipid metabolism. PMID- 10680059 TI - Hyperlipidaemia and cardiovascular disease. PMID- 10680060 TI - What's sex got to do with it? PMID- 10680061 TI - Therapy and clinical trial. PMID- 10680062 TI - Monoclonal antibody against tetrahydrocannabinolic acid distinguishes Cannabis sativa samples from different plant species. AB - The cross-reaction of anti-delta 1-THCA MAb against other cannabinoids was very wide. However, other naturally occurring and synthetic phenolics including opium alkaloids did not react to the MAb. Using this ELISA, this paper reports application of the competitive ELISA for detection of marijuana samples. The ELISA described here was very sensitive to the ether extracts of marijuana samples when compared to those of other plants. The assay provided a sensitive method useful for the judge of marijuana samples. PMID- 10680063 TI - Screening experiments of ecstasy street samples using near infrared spectroscopy. AB - Twelve different sets of confiscated ecstasy samples were analysed applying both near infrared spectroscopy in reflectance mode (1100-2500 nm) and high performance liquid chromatography (HPLC). The sets showed a large variance in composition. A calibration data set was generated based on the theory of factorial designs. It contained 221 N-methyl-3,4-methylenedioxyamphetamine (MDMA) samples, 167 N-ethyl-3,4-methylenedioxyamphetamine (MDE), 111 amphetamine and 106 samples without a controlled substance, which will be called placebo samples thereafter. From this data set, PLS-1 models were calculated and were successfully applied for validation of various external laboratory test sets. The transferability of these results to confiscated tablets is demonstrated here. It is shown that differentiation into placebo, amphetamine and ecstasy samples is possible. Analysis of intact tablets is practicable. However, more reliable results are obtained from pulverised samples. This is due to ill-defined production procedures. The use of mathematically pretreated spectra improves the prediction quality of all the PLS-1 models studied. It is possible to improve discrimination between MDE and MDMA with the help of a second model based on raw spectra. Alternative strategies are briefly discussed. PMID- 10680064 TI - Origin of blood ethanol in decomposed bodies. AB - Problems related to blood contamination by other postmortem fluids in decomposed bodies (DB) make the interpretation of medicolegal blood alcohol levels (B EtOH) a very difficult task. So the aim of this paper is to show the utilization of vitreous humor (VH) as the biological fluid for an unequivocal determination of ethanol origin in DB for forensic purposes. Alcohol was determined in VH, blood (chest fluid-CF) and urine (Ur) collected from 27 DB in different states of putrefaction. A simple head-space gas-chromatographic method was used. In fifteen cases alcohol was found to be of endogenous production due to its absence in VH. In the twelve remainders, alcohol was detected in VH and CF in an atypical distribution. Examining the reliable scene and historical information together with the analytical data, ethanol origin in these cases was classified: endogenous production (3 cases), ingested (2 cases), both (2 cases), contaminated plus endogenous production (3 cases) and unable to determine (2 cases). According to the results obtained it was possible to conclude that alcohol analysis in VH is fundamental for determining the origin of ethanol detected in CF of DB. PMID- 10680065 TI - Robustness of the Y STRs DYS19, DYS389 I and II, DYS390 and DYS393: optimization of a PCR pentaplex. AB - Various technical methods were investigated with the aim of developing a multiplex system to amplify five Y-chromosome STR loci in the same PCR reaction: DYS393, DYS19, DYS390, DYS389 I and DYS389 II. A sequenced allelic ladder was constructed with previously sequenced alleles including the most common ones. A number of reamplification conditions of the allelic ladders were tested. The pentaplex was evaluated for typing using two different platforms (ABI and ALF) with promising results. However, in degraded samples non-specific artifacts were observed in the DYS393 system in the same range of sizes as the real alleles. This system can also be typed in females under relatively low stringency conditions in the PCR amplification, making this system prone to errors in critical samples. This lack of specificity can be reduced by increasing the stringency of the PCR conditions. The DYS19 ladder cannot be reamplified as stutters appear after a few reamplifications. These stutters are probably due to a 2 bp slippage induced by the presence of a TA repeat stretch in the PCR amplified fragments. Non-specific products were also noted in the DYS389 I and DYS389 II amplification, although out of the range of other alleles in this pentaplex. This newly constructed pentaplex has proved to be very useful in population genetic studies because all five Y STR markers can be loaded in the same lane of a gel with other Y STR singleplex or multiplexes. The usefulness of Y-chromosome STRs in criminal casework is especially evident in analyzing azoospermic individuals. PMID- 10680066 TI - Drugged driving in the Nordic countries--a comparative study between five countries. AB - The purpose of this study was to compare whether the high incidence of drugged driving in Norway was different to that in the other Nordic countries. All blood samples received by Nordic forensic institutes during one week in 1996, from drivers suspected by the police of driving under the influence (Denmark: n = 255, Finland: n = 270, Iceland: n = 40, Sweden: n = 86, Norway: n = 149), were analysed for alcohol and drugs (benzodiazepines, cannabinoids, amphetamines, cocaine, opiates and a number of antidepressant drugs) independent of the primary suspicion, and using the same analytical cut-off levels at the different institutes. The primary suspicion was directed towards drugs in more than 40% of the Norwegian cases, drugs were detected in more than 70% of these samples. In only 0-3% of the cases from Denmark, Finland and Iceland, were drugs suspected, while the corresponding frequency for Sweden was 17%. However, evidential breath analyses were used for about three-quarters of the Swedish drivers suspected to be influenced by alcohol. Blood alcohol concentrations (BAC's) below the legal limits were found in 32, 18 and 2% of the Norwegian, Icelandic and Finnish cases, respectively (BAC < 0.05%), in 10% of the Danish cases (BAC < 0.08%) and in 20% of the Swedish cases (BAC < 0.02%). Drugs were most frequently found in the Norwegian and Swedish cases with no alcohol (80-83%). Similar frequencies of drugs in samples with BAC's above the legal limits (19-22%), were obtained for all countries. Benzodiazepines, tetrahydrocannabinol and amphetamine represented the most commonly detected drugs. Our results show that differences between Norway and other Nordic countries with regard to drugs and driving, are connected to the selection criteria made by the police and with more focus on drugged driving in Norway. PMID- 10680067 TI - Colchicine poisoning by accidental ingestion of meadow saffron (Colchicum autumnale): pathological and medicolegal aspects. AB - Although intoxications with colchicine, the alkaloid of Colchicum autumnale (meadow saffron), are well known, in most cases the intoxications are evoked by oral or parenteral preparations traditionally used as medication against gout. The accidental ingestion of Colchicum autumnale, on the other hand, is a rare event and has to our knowledge only twice been described in detail. We report a further case in which two persons confused this highly poisonous plant with wild garlic (Allium ursinum), a popular spice in the Central European cuisine. While one person merely complained about a 3-day episode of nausea, vomiting and watery diarrhea, the second person died of multi-organ system derangements 48 h after the ingestion of the colchicum leaves. At autopsy hemorrhagic lung oedema, hypocellular bonemarrow, centrilobular fatty necrosis of the liver and necrosis of the proximal convoluted tubuli of the kidneys were observed. A colchicine concentration of 7.5 micrograms/ml was found in the bile whereas no substance was detected in the postmortem blood. PMID- 10680068 TI - Cancer and aging. An evolving panorama. AB - This article illustrates how the nosology of cancer evolves with the patient's age. If the current trends are maintained, 70% of all neoplasms will occur in persons aged 65 years and over by the year 2020, leading to increased cancer related morbidity among older persons. Cancer control in the older person involves chemoprevention, early diagnosis, and timely and effective treatment that entails both antineoplastic therapy and symptom management. These interventions must be individualized based on a multidimensional assessment that can predict life expectancy and treatment complications and that may evaluate the quality of life of the older person. This article suggests a number of interventions that may improve cancer control in the aged. Public education is needed to illustrate the benefits of health maintenance and early detection of cancer even among older individuals, to create realistic expectations, and to heighten awareness of early symptoms and signs of cancer. Professional education is needed to train students and practitioners in the evaluation and management of the older person. Of special interest is the current initiative of the Hartford Foundation offering combined fellowships in oncology and geriatrics and incorporating principles of geriatric medicine in medical specialty training. Prudent pharmacologic principles must be followed in managing older persons with cytotoxic chemotherapy. These principles include adjusting the dose according to the patient's renal function, using epoietin to maintain hemoglobin levels of 12 g/dL or more, and using hemopoietic growth factors in persons aged 70 years and older receiving cytotoxic chemotherapy of moderate toxicity (e.g., CHOP). To assure uniformity of data, a cooperative oncology group should formulate a geriatric package outlining a common plan for evaluating function and comorbidity. This article also suggests several important areas of research items: Molecular interactions of age and cancer Host-tumor interactions in the older tumor host Chemoprevention of cancer and aging Laboratory evaluation of aging Development of shorter forms of geriatric assessment Management of the frail cancer patients Clinical trials of tumor-specific issues. PMID- 10680069 TI - Aging and cancer in America. Demographic and epidemiologic perspectives. AB - It has been stated in this article and elsewhere that cancer patients aged 65 years and older deserve special attention as a target group for research efforts across the cancer-control spectrum. The available data show that the vulnerability of older persons to cancer is unmistakable. Clinicians will be treating more older patients as the nation ages. The future needs of this segment of the population must be anticipated. In this context, the following generic treatment questions are pertinent. What are the peculiarities of the aged host of which clinicians must be aware in evaluating the older cancer patient? Do various forms of cancer present differently in the elderly? How can be complications caused by the multiple pathologies inherent in the older patient be anticipated? What are the potential hazards and limitations of surgery, radiotherapy, and chemotherapy for older persons with cancer? What is known regarding increased risk of adverse reactions to medications, drugs, and interaction of drugs in older patients? The surveillance data and population estimates and projections presented in this article illustrate the extent of the problems of cancer in the elderly at the macro level. For the individual patient, the special knowledge of aging individuals and their health status based on geriatric medicine and gerontology that has been accumulating for the past several decades needs to be incorporated into the oncology armamentarium that has developed during the same period. The information and expertise from both fields must converge, and new knowledge must be developed at the aging/cancer interface and applied for the optimal treatment of cancer in the elderly. PMID- 10680070 TI - Molecular interactions of cancer and age. AB - The authors believe that the aging process--the loss of youthful resilience--is caused by the decline of many hormones. By restoring these hormone levels, the decay associated with old age can be eliminated and in some cases, perhaps reversed. The hormones that naturally occur in the body should be replenished through a medically sound regimen. The hormones must work together. The data indicate that aging occurs at two levels: systemically and cellularly. Systemic controls regulate the rates of intracellular enzymatic processes that accelerate to protect against aging. PMID- 10680071 TI - Hematopoiesis and cytokines. Relevance to cancer and aging. AB - Impaired hematopoiesis and dysregulated cytokine expression have important implications for cancer in the elderly. In aged people, hematopoiesis is dysregulated and becomes paradoxically down-modulated under periods of increased hematopoietic demand. This down-modulation may explain, at least in part, the increased incidence of anemia in the elderly, although the cause of anemia can usually be identified in these patients and frequently reversed with targeted therapy. An age-associated decrease in the expression of interleukin-2 may contribute to impaired cellular immunity. Additionally, the increased interleukin 6 production frequently found in the elderly may participate in promoting the survival and proliferation of malignant myeloma and in inducing resistance by myeloma cells to therapies that act through apoptosis. Dysregulation of the expression of these and other cytokines may be a mechanism contributing to age related impairment of the hematopoietic response, the genesis and therapeutic resistance of specific malignancies, and cancer cachexia. PMID- 10680072 TI - Assessment of the older cancer patient. AB - The correct assessment of a cancer patient is a key step in the treatment process. In older people, this assessment entails not only the patient's basic medical history and the standard cancer staging, but also much more comprehensive evaluation of the various facets of the patient's health and environment that may interfere with his or her therapy. Patient fitness for elective surgery, radiation therapy, and chemotherapy must be considered. Geriatricians have defined the relevant aspects of the general evaluation of the older person, and now this work is being adapted to cancer patients. This article reviews the various aspects of a comprehensive assessment applicable to the cancer patient in settings such as academic oncology programs, cooperative group studies, and private oncology practice. PMID- 10680073 TI - Qualitative research for the study of cancer and age. AB - Qualitiative research emphasizes identification, illumination, and understanding of phenomena, the meaning and theory behind which are unpresumed by the investigator. Although quantitative techniques are used to test predetermined hypotheses, qualitative techniques are used to generate hypotheses. Qualitative techniques have only begun to be used in medical research in the past decade but are especially useful in exploring content areas about which little is known and in eliciting and understanding the patient's perspective. Despite the aging of the United States population, the cancer illness experience has not been well studied in older patients. Because communication preferences, treatment decision making styles, psychosocial issues, and the illness experience itself may be significantly different for older persons diagnosed with cancer than for younger persons, qualitative research techniques can be used to identify those differences critical to the effective health care of this burgeoning population. PMID- 10680074 TI - Cancer screening in the elderly population. AB - This article reviews the current state of knowledge regarding cancer screening in the geriatric population. Care of the elderly requires knowledge of underlying physiologic changes, comorbidities, quality-of-life factors, and life expectancies. There is always the danger that ageism may prevent elderly cancer patients from receiving the proper treatment. On the other hand, overzealous treatment can lead to adverse results if elderly patients are not properly targeted based on current evidence of the benefits and risks of specific screening practices. PMID- 10680075 TI - Chemoprevention of breast cancer in the older patient. AB - Age is the most important risk factor for the development of breast cancer. The risk of breast cancer continues to increase in American women until the age of 80 years. A family history of breast cancer helps identify those who possibly have the highest risk of developing breast cancer; however, most women who develop breast cancer do not have a first-degree relative with a history of breast cancer. Currently, the Gail model is a commonly used model to identify risk, and this model has now been validated in several populations of women undergoing screening for breast cancer. The first large-scale breast cancer prevention trial investigating the preventive effects of tamoxifen has demonstrated a decrease in the development of breast cancer by almost 50% in the women in the tamoxifen treatment arm as compared with those receiving placebo. The NSABP P-1 trial was the largest of the three tamoxifen breast cancer prevention trials and had the greatest power to detect a difference between the two treatment groups in breast cancer events. This trial also included the largest percentage of postmenopausal women. It is unclear why the Italian and Royal Marsden Hospital trials had negative results regarding the preventive effects of tamoxifen. These two trials were strikingly different from the NSABP P-1 trial, however, and they included a different population of women. The issues surrounding the use of HRT for treatment of hot flashes in the Italian and Royal Marsden Hospital trials adds to the controversy concerning the negative results of these trials. The new SERM, raloxifene, has shown promise in preliminary studies as a preventive agent for breast cancer. The STAR trial will open soon and will evaluate the efficacy of raloxifene in preventing breast cancer in a prospective fashion, comparing its efficacy with tamoxifen treatment. Other endpoints will evaluate side effects such as menopausal symptoms, endometrial cancer, thromboembolic events, and benefits regarding serum lipids and incidence of osteoporotic bone fractures. The development of SERMs results from an understanding of novel mechanisms of ER modulation and allows targeting for favorable effects in specific tissues. The challenge is to develop an ideal SERM that is effective in preventing breast cancer and does not increase the risk of endometrial cancer, while providing beneficial estrogenic effects on serum lipids and bone mineral density changes. Estrogen receptor-mediated intracellular processes are complex. There are at least two different types of estrogen receptors. The alpha receptors predominate in the breast and uterus, and the beta receptors predominate in the bone and blood vessels. Many proteins also interact with these receptors as co-activators or co-repressors. Transcription-activating factors modulate the effects of estrogen on its target genes. Future prevention strategies may use a combined targeted approach to inhibit ER-mediated cancer progression pathways. The retinoids are under investigation in prevention studies for a multitude of cancers, because they have been shown to inhibit cellular proliferation and to induce cellular differentiation. The retinoid 4HPR was selected for use in breast cancer prevention studies because of its low toxicity profile and prevention efficacy in preclinical studies. It is now being used in combination with tamoxifin in a phase II breast cancer prevention trial. Multiple surrogate endpoint biomarkers are being measured before and after treatment, including measurement of serum IGF-I levels. Future directions in breast cancer prevention include the development of more potent hormonal therapies that completely inhibit ER-mediated cancer progression and, ultimately, multitargeted therapies involving agents that work synergistically. PMID- 10680076 TI - Radiation therapy of the older patient. AB - Radiotherapy has a major role in the multidisciplinary approach to cancer therapy. It is widely used for curative and palliative treatment of cancer involving various sites. Radiotherapy is of particular benefit to older and frail cancer patients as an alternative to surgery and to systemic therapy. The available data on the sensitivity of normal tissues to radiotherapy in elderly patients strongly suggest that older patients with good functional status tolerate radiotherapy as well as younger patients and have comparable tumor response and survival rates. Aggressive radiotherapy should not be withheld from older patients because of chronological age alone. PMID- 10680077 TI - Cancer surgery in the elderly. AB - The incidence of most cancers increases with age. Although the risk for surgery increases in elderly patients who have comorbidities, evaluations of risk can allow interventions that may decrease morbidity and mortality. Appropriate treatments should be offered to the elderly until studies demonstrate the elderly can safely be managed differently from younger patients. The elderly should not be denied adequate treatment simply because of age. PMID- 10680078 TI - Antineoplastic chemotherapy of the older cancer patient. AB - Cancer chemotherapy may be effective and safe in older patients if some proper provisions are made. Doses of chemotherapy should be adjusted to the patient's glomerular filtration rate, and his or her hemoglobin should be maintained for the duration of the therapy. For patients who are 70 years of age or older and who are receiving moderately toxic chemotherapy, growth factors should be used. The risk of mucositis increases with the age of the patient, so it is important to treat it aggressively at the first signs of the complication. PMID- 10680079 TI - Breast cancer and aging. Clinical interactions. AB - The incidence and mortality rates of breast cancer increase with age. As the geriatric population grows, the number of breast cancer cases will reach epidemic proportions. The number of coexisting medical conditions also increases with advancing age. The presence and severity of comorbid conditions influences an individual's ability to tolerate procedures and treatments and must be considered in making disease-management decisions. Screening mammography can potentially save lives in older women. Women whose life expectancy exceeds 5 years should continue annual screening mammography. Choices for local definitive therapy, systemic adjuvant therapy, and treatment of metastatic disease should be based on patient preference and ability to tolerate the planned procedure. In general, otherwise healthy older women should be offered the same treatment options given to younger, postmenopausal women. Alternative, less aggressive, or nonstandard approaches are warranted in women whose life expectancy is limited or who are unable or unwilling to undergo standard management procedures. PMID- 10680080 TI - Cancer in the frail patient. A coming epidemic. AB - With the aging of the population, frailty has emerged as a new clinical entity. The frail person has exhausted any functional reserve. Current criteria for the recognition of frailty include age of over 85 years, dependence in one or more activities of daily living, three or more comorbid conditions, and the presence of one or more geriatric syndromes. It is calculated that there are approximately 6 million frail patients in the United States and approximately 400,000 of them have cancer. Management of cancer in the frail person is mainly comprised of palliation, which may include some forms of chemotherapy, such as navelbine, gemcitabine, or low-dose taxanes. PMID- 10680081 TI - Acute myelogenous leukemia and aging. Clinical interactions. AB - Effective treatment of the elderly patient with AML remains a challenging task. Acute myelogenous leukemia is clearly a different disease in the elderly than in the young, for many reasons, both clinical and biologic, which contribute to the worse prognosis in the elderly. The elderly, as a group, have been underrepresented in clinical trials. Several important prognostic variables have been identified and described, however, that can help the physician select the appropriate treatment for any individual patient. Age itself should not preclude an attempt at therapy, especially for AML, which progresses very rapidly in the absence of treatment. After careful analysis of prognostic factors, in any individual patient, however, the outlook may be so poor that it may be desirable to withhold treatment. With a better understanding of the pathophysiology of AML in the elderly, more targeted and less toxic treatment regimens will become available. At present, however, clinicians must use an improved understanding of the disease to predict its behavior in an individual patient, so that the currently available treatment modalities are used most prudently. PMID- 10680082 TI - The family caregiver of the older cancer patient. AB - Cancer care is becoming increasingly complex. The health care practices of the present day make it imperative that the family provide assistance in the day-to day management of a patient's symptoms and in the implementation of home care. This increased responsibility for the family caregiver can come at great cost to the overall functioning of the family, because of role changes, changes in the family structure, and financial stressors, among others. Caregivers can experience adverse effects on their mental and physical health that can remain long after the caregiving role has ended. Although much of the research has focused on the negative elements of caregiving, it must be remembered that there are positive aspects to caregiving, which are now receiving attention in the research literature. Although other factors affect caregiving, the model of factors important in the caregiving experience presented in this article may provide the impetus for studying the relationships among these different factors. Results of these studies will allow the development of specific interventions to help caregivers be more effective in their caregiving role and, at the same time, address the overall impact of caregiving on the family. PMID- 10680083 TI - Clinical research in the older cancer patient. AB - The opportunities for conducting focused research in older patients with cancer continue to increase and reflect the increasing awareness of the incidence of cancer in older persons. As the population ages, cancer will become more and more a problem of the elderly. Despite the magnitude of this problem, exploration of the unique problems associated with cancer in the aging has only begun. Economic issues, access to care, and the frequency of other serious illness in older patients often make screening and treatment of malignancy in older patients a great challenge. Oncologists and geriatricians must work together to guarantee that appropriate funding is made available for cancer research in the elderly population. PMID- 10680084 TI - Association of Helicobacter pylori with gastroduodenal diseases. AB - Helicobacter pylori was first cultured in vitro in 1982. This bacterium is a spiral gram-negative rod which grows under microaerophilic conditions. The ecological niche is the mucosa of the human stomach which had been thought to be aseptic before the discovery of this bacterium. This organism causes a long lasting infection throughout a person's life if there is no medical intervention. Numerous persons are infected with the organism around the world, and the rate of infection in Japan is nearly 50% of the population. However, the route of infection remains unclear because the organism has not been isolated from any environment other than several animals. H. pylori is now recognized as a causative agent of gastritis and peptic ulcers. Though gastritis, and especially chronic active gastritis, is observed at least histologically in all persons with H. pylori, peptic ulcers develop in only some infected persons. Specific factors in the host and/or the bacteria are needed for the development of peptic ulcer disease. Furthermore, H. pylori is considered to be related to the development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma, especially those of low grade. Also, H. pylori infection is a major determinant for initiating the sequence of events leading to gastric cancer. In some patients with low-grade gastric MALT lymphoma, the eradication of H. pylori led to a regression of lesion. Gastric cancer has been induced in Mongolian gerbils with long-term H. pylori infection. The combinations of drugs, which consist of an antisecretory agent (acid-suppressing agent) and antimicrobial agents, are used for the eradication of the organism. Eradication therapy is recommended at least for patients with peptic ulcers. PMID- 10680085 TI - Multiple sclerosis and measles virus. AB - Epidemiological studies suggest that patients with multiple sclerosis (MS) are exposed to some infectious agent(s) before puberty. The presence of virus-induced demyelination in animal models indicates that demyelination can occur following the trigger of a virus infection. Data regarding the immunological abnormalities to measles virus (MV) and the presence of neurological complications induced by MV infection suggest that MV may be a causative agent of the demyelination observed in MS. Numerous virological studies (e.g., morphological observation, virus isolation, and the search for the MV gene) have been performed, though definite evidence identifying MV as the causative agent has not yet been obtained. PMID- 10680086 TI - Role of human parvovirus B19 in the pathogenesis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease of unknown origin. It has been speculated that infectious agents are responsible for triggering RA. Persistent infection with human parvovirus B19 and its induction of immunopathology are hypothesized to explain the initiation and perpetuation of the disease process. PMID- 10680087 TI - Pediatric admission for vaccine preventable diseases: a 5-year survey from 1994 to 1998 in Aichi Prefecture. AB - Several infectious childhood diseases can be prevented by vaccination. A survey of hospital admissions for such diseases was conducted in Aichi Prefecture over 5 years beginning in 1994. A questionnaire was sent annually to hospitals with 100 or more beds to obtain information on pediatric patients requiring hospitalization for 10 vaccine preventable diseases. Information was obtained on 3,953 patients. Most admissions were for measles (49%), followed by mumps, chickenpox, pertussis, rubella, and tuberculosis. Over half of the patients were under 3 years old, with 20% aged under 12 months, 25% aged from 12 months to 2 years, and 10% aged from 2 to 3 years. The average hospital stay was longest for tuberculosis and pertussis, and was around 1 week for the other diseases. Familial transmission was the most common source of infection identified. The only death was an unvaccinated patient with measles pneumonia. Sequelae were reported at the time of discharge in 15 patients (0.4%), and were permanent in some cases. Some 96% of the patients surveyed were unvaccinated against the disease causing hospitalization. The fact that there were 14 patients with sequelae and the one patient who died were unvaccinated, emphasizes the need to promote vaccination. PMID- 10680088 TI - Plasmodium infection-induced changes in salivary gland proteins of malaria vector Anopheles stephensi (Diptera:Culicidae). AB - Parasitism by Plasmodium yoelii yoelii induced 18 polypeptides in the salivary glands of aging malaria vector Anopheles stephensi. A polypeptide of low molecular size (30 kDa) could generally be induced at all infected stages. On day 5 post blood feeding (PBF), no new polypeptide could be found in the salivary glands. Seven polypeptides of low molecular size and 3 of high molecular size could be induced on day 11 PBF, which inducibility coincided with the invasion of the salivary glands by the sporozoites. Quantitatively, soluble proteins decreased in the salivary glands by about one-third in females that had consumed infected or uninfected blood meal on day 9 (oocysts stage) as compared to nonfeeding females. However, on day 15, in the salivary glands invaded by sporozoites, the amount of proteins obtained from infected females was approximately 26% lower than that obtained from uninfected females. A similar reduction was also observed in aged (20 days PBF) salivary glands of infected mosquitoes. These proteins could confer parasite tolerance to the females and enhance parasite transmission potential. PMID- 10680089 TI - Non-radioactive screening of DNA polymerases by using oligo r(A-U) as primer/template for reverse transcriptase activity. PMID- 10680090 TI - Salmonella Oranienburg involved in a variety of diseases. PMID- 10680091 TI - Apparent cure of hepatitis C virus carrier state after hepatitis B virus superinfection. PMID- 10680092 TI - Selection of methicillin-resistant Staphylococcus aureus following increased use of vancomycin despite restricted use of third generation cephem. PMID- 10680093 TI - Review of the effect of spiritual and religious factors on mortality and morbidity with a focus on cardiovascular and pulmonary disease. PMID- 10680094 TI - The Toronto Cardiac Rehabilitation and Secondary Prevention Program: 1968 into the new millennium. AB - Given our approach to the cardiac rehabilitation process, which is reflected in the program structure and services and our high patient volume, this program model is effective for us. The model permits us to treat relatively large number of patients with relatively small numbers of staff. On average, a patient attends 32 supervised exercise sessions at the Centre over the course of 12 months. This is actually fewer supervised sessions than the popular model of 3 times per week for 12 weeks. However, the 12-month program provides an additional 9 months to work with patients on heart-healthy lifestyle modifications. At the same time, we realize our model is not the model of choice for all people in all settings for a variety of reasons. We trust that some elements of our program may be of interest and beneficial to some readers. Undoubtedly, the program will continue to evolve and develop into the future. Currently, we are conducting a cardiac rehabilitation outcomes study in an effort to determine the appropriate duration of cardiac rehabilitation to achieve optimal physiological, psychological, and cost benefits for patients. This study involves more than 700 patients and the results are intended to help us further refine the program structure and selected program elements. As the new millennium approaches, healthcare system reforms and continuing changes in the delivery of medical care to cardiac patients present opportunities, challenges, and some uncertainties for cardiac rehabilitation. To continue our services to patients and the medical community, cardiac rehabilitation programs will need to identify and develop even more innovative and effective concepts in response to ever-changing local, regional, and national issues. PMID- 10680095 TI - Rationale and design of the national emphysema treatment trial. A prospective randomized trial of lung volume reduction surgery. The national emphysema treatment trial research group AB - The National Emphysema Treatment Trial is a multicenter, randomized clinical trial of medical therapy vs medical therapy plus lung volume reduction surgery (LVRS) for the treatment of patients with severe bilateral emphysema. LVRS will be accomplished by bilateral stapled excision via median sternotomy or video assisted thoracoscopic surgery. Every patient will complete 6 to 10 weeks of pulmonary rehabilitation prior to randomization and will participate in a maintenance program of pulmonary rehabilitation after randomization. The primary outcome to be assessed by the trial is survival. Additional outcomes to be assessed are maximum exercise capacity, pulmonary function, oxygen requirement, distance walked in 6 min, quality of life, respiratory symptoms, and health-care utilization and costs. In addition, selected clinics will evaluate lung mechanics and respiratory muscle function, partial and maximal flow-volume curves, gas exchange during maximal exercise, and right heart function. The trial is targeted to enroll patients with severe emphysema who have no significant comorbid conditions; each patient will be randomized to one of the two treatment groups. The study duration is 4.5 years with a close-out period of 6 months. PMID- 10680096 TI - A comparison of autogenic drainage and the active cycle of breathing techniques in patients with chronic obstructive pulmonary diseases. AB - PURPOSE: The effects of a long-term treatment of autogenic drainage (AD) and the active cycle of breathing techniques (ACBT) were evaluated in patients with chronic obstructive pulmonary disease (COPD). METHODS: Thirty clinically stable male COPD patients were randomly assigned to AD or the ACBT treatment for a 20 day treatment period. Patients were assessed through pulmonary function tests, arterial blood gases, a 6-minute walking test, and a modified Borg Scale before, and immediately after the walking test. RESULTS: Autogenic drainage improved forced vital capacity, forced expiratory volume in 1 second, peak expiratory flow rate, forced expiratory volume from 25 to 75%, chronic hypercapnia, arterial oxygenation, exercise performance, and dyspnea perception during exercise. The ACBT increased forced vital capacity, peak expiratory flow rate, arterial oxygenation and exercise performance. Peak expiratory flow rate increased in AD more than in ACBT. In AD treatment, the increase in oxygen saturation was significantly higher than in ACBT treatment. Chronic hypercapnia improved significantly in AD treatment than in ACBT. No differences were found in other lung function parameters. CONCLUSIONS: Autogenic drainage is as effective as the ACBT in cleaning secretions and improving lung functions. These techniques can be used in stable COPD patients according to the patients' and the physiotherapists' preferences. PMID- 10680097 TI - Muscle metabolism assessed by phosphorus-31 nuclear magnetic resonance spectroscopy after myocardial infarction in rehabilitated patients: a 1-year follow-up. AB - BACKGROUND: The most common effect of postmyocardial infarction (post MI) rehabilitation is an increase of peak maximal oxygen consumption correlated with changes in calf muscle metabolism, but there are few data on follow-up after rehabilitation on skeletal muscle and maximal oxygen consumption. The purpose of this study was to investigate the respective modifications in skeletal muscle metabolism and peak oxygen consumption (VO2) occurring during a supervised rehabilitation program and 1 year after MI in patients free of heart failure. METHODS: Fifteen outpatients were studied prospectively after the acute phase of the MI, at the end of the rehabilitation program (2 months after the MI), and 1 year after. The rehabilitation comprised 20 sessions with three sessions per week. The program consisted of exercise training with bicycle, arm ergometer, and treadmill. The program also included respiratory exercises, psychological support, and counseling for secondary prevention of cardiovascular diseases. At each visit, a stress test on a bicycle ergometer was performed and the peak VO2 was measured. Phosphorus magnetic resonance spectroscopy of the gastrocnemius muscle was performed at rest and during a plantar flexion-type exercise against an adjustable load. Data were analyzed using analysis of variance and post-hoc test when appropriate. RESULTS: The mechanical power output measured during the bicycle exercise increased from 111 +/- 28 watts at the post MI test to 136 +/- 40 watts after rehabilitation (post rehab) and decreased to 125 +/- 36 watts at 1 year. The peak VO2 increased significantly (P < 0.05) from 22 +/- 7 ml/kg-1/min-1 (post MI) to 27 +/- 9 ml/kg-1/min-1 (post rehab), and decreased significantly to 24 +/- 8 ml/kg-1/min-1 (1 year). The mechanical power output measured in the magnet during the stress test increased from 2.22 +/- 0.13 watts (post MI) to 2.85 +/- 1.24 (post rehab), and stabilized at 2.78 +/- 1.10 watts at 1 year. At the highest workload attained in the three successive tests, the phosphocreatine/(phosphocreatine + inorganic phosphate) ratio rose significantly (P < 0.05) from 0.46 +/- 0.13 (post MI) to 0.51 +/- 0.13 (post rehab) and remained at 0.51 +/- 0.13 at 1 year. CONCLUSION: The improvement of the peak VO2 after training post MI is not maintained 1 year later. This decline is not accompanied by muscular metabolic abnormalities. This suggests that the muscle metabolism after MI remains normal, and that the long-term decrease of the peak VO2 reflects a global deconditioning that should be avoided by maintaining a long term phase III rehabilitation program. PMID- 10680098 TI - A home-based maintenance exercise program after center-based cardiac rehabilitation: effects on blood lipids, body composition, and functional capacity. AB - BACKGROUND: Previous research indicates that patients exiting a 12-week cardiac rehabilitation program (CRP) have difficulty maintaining an adequate exercise program. Thus, the authors' purpose was to determine if a home-based exercise program would enable patients to maintain/improve their blood lipids, body composition, and functional capacity after exiting the CRP. METHODS: Thirty-one patients exiting an initial 12-week CRP were assigned randomly to the home-based (HB) intervention or the standard care (SC) condition. After one home visit, the HB participants (n = 16) were contacted by telephone every other week by CRP staff and completed and returned weekly exercise logs. The SC participants (n = 15) had no contact with the CRP other than to schedule follow-up tests. A third group (n = 17), randomly selected from patients that elected to remain in the center-based CRP (CB) for the same duration, also were examined. All groups underwent exercise testing, fasting blood lipid analysis, and body composition assessment before starting CRP (0M), after 3 months (3M) in a standard CRP, and after 9 months (12M) in either HB, SC, or CB condition (12 months after starting CRP). RESULTS: Analysis of variance indicated that there were significant increases in metabolic equivalents and high-density lipoprotein, in all three groups, over time. However, analysis of covariance revealed no significant differences between the HB, SC, and CB groups at 12M for any variable. CONCLUSIONS: These data indicate that the HB program was as effective as the CB program at improving/maintaining functional capacity, blood lipids, and body weight/composition. The similar success of the SC group is likely due to their prior experience in CRP and knowledge of follow-up testing. Home-based maintenance program could be offered as a low-cost alternative to CB programs. PMID- 10680099 TI - Contemporary cardiac rehabilitation: patient characteristics and temporal trends over the past decade. AB - BACKGROUND: Recent and dramatic therapeutic advances, aggressive care of the elderly, and a heightened awareness of secondary prevention have had a significant, albeit incompletely described, impact on cardiac rehabilitation (CR) programs. The authors did a retrospective analysis on 1,848 patients from their phase II CR program that were treated over the past decade. The authors sought to identify and analyze how advances in cardiovascular care might be related to temporal changes in demographics, medical comorbidities, coronary artery disease risk factors, and cardiovascular history among individuals with cardiac disease who have completed the phase II CR program over a 10-year period. METHODS: Cardiac rehabilitation records were reviewed from 1986 to 1996 at Akron City Hospital. The data were compiled prospectively by nurses and exercise physiologists and were subsequently analyzed. The charts reviewed were from 1,848 patients who completed outpatient phase II CR. RESULTS: The program began in 1986 with 53 patients completing CR and evolved to 309 in 1996. There has been an increase in the number of elderly patients referred to and completing the program. The number of participants older than 65 years of age increased from 28.3% in 1986 to 52.1% in 1996. Cardiac rehabilitation participants reflect the known demographics of patients with clinical coronary artery disease. Men outnumber women and, on average, the female participants are older than the male participants. There has been a statistically significant increase in medical comorbidities over the course of the study. Although the absolute number of patients entering CR after coronary artery bypass graft survey has remained fairly constant, there has been a dramatic increase in the absolute number of patients entering CR after percutaneous tansluminal coronary angioplasty (with the latter recently comprising a majority of CR participants). CONCLUSION: There is a significant trend in the attendance and completion of CR programs by older individuals, which suggests a greater awareness of patients and their physicians. The growth of the program is fueled by high-risk patients with more comorbidities who potentially are the group of patients able to obtain the greatest absolute benefit from CR. PMID- 10680100 TI - Mood-altering drugs and alternative integrative medicine: what we don't know, what we need to know, and how we can find out. PMID- 10680101 TI - The self-reporting of psychiatric medications in patients scheduled for elective surgery. AB - STUDY OBJECTIVE: To determine by survey the percentage of patients, over the age of 21 years, reporting for elective surgery who also are taking psychotropic medications. DESIGN: Institution-approved, anonymous survey. SETTING: Teaching hospital. PATIENTS: 169 patients who were scheduled for elective surgery. INTERVENTIONS: After completion of the preoperative evaluation by members of the anesthesia care team, all patients were given an institution-approved survey of medications they were taking in addition to their nonpsychiatric medications. The survey listed 33 drugs known to affect central nervous system neurotransmitters, and included the most commonly prescribed antidepressants, antipsychotics, benzodiazapines, and lithium. Over-the-counter drugs known to affect mood, such as melatonin, also were included in the survey. Patients were not asked the indications for the medications, and no psychiatric questions were asked of the patients. MEASUREMENTS AND MAIN RESULTS: 300 surveys were distributed, and 169 patients completed the survey for a response rate of 53%. Forty-three percent of all patients who completed the survey admitted to taking one or more of the psychotropic medications. Of these patients, 35% were taking antidepressants, 34% were taking benzodiazapines, 19% were taking combination therapies, and 11% took antipsychotics, lithium, or over-the-counter drugs such as melatonin. CONCLUSION: The number of patients taking psychotropic medications and who present for elective surgery is high. The anesthetic implications of this drug usage are essentially unknown. Additional study of these medications and their impact on anesthetic care is warranted. PMID- 10680102 TI - Course of molecular hemostatic markers during and after different surgical procedures. AB - STUDY OBJECTIVE: To establish the most vulnerable time of thrombi formation with regard to the plasmatic (noncellular) part of the coagulatory and fibrinolytic systems. DESIGN: Nonrandomized observational study. SETTING: A surgical and an orthopedic unit and the central laboratory of a university hospital. PATIENTS: 61 consenting ASA physical status I and II inpatients undergoing four different types of surgery: total hip replacement (THR): 16 patients; hemicolectomy: 15 patients; endoscopic cholecystectomy: 15 patients; subtotal thyroid resection: 15 patients. INTERVENTIONS: The time course of 11 procoagulatory and fibrinolytic parameters was examined during the different types of surgery. Blood samples were drawn on the day before surgery, directly before the induction of general anesthesia, 1 to 2 hours postoperatively, and on the mornings of postoperative days 1, 2, 3, 4, and 5. MEASUREMENTS AND MAIN RESULTS: The coagulation samples were centrifuged within 1 hour of collection at 2,300 g for 15 minutes at 4 degrees C. Hemoglobin, hematocrit, platelets, fibrinogen, prothrombin time, activated partial thromboplastin time, thrombin time, antithrombin III, and protein C were determined immediately on laboratory arrival of the samples. The samples were aliquoted at -70 degrees C. They were thawed within 2 weeks and prepared for the following assays: thrombin-antithrombin III complexes (TAT complexes), D-dimers, and plasminogen activator inhibitor type 1. Maximum activation of coagulation is not reached until 2 hours postoperatively and slowly decreases until normal values are reached around the fifth postoperative day. Parameters displaying the greatest changes are TAT-complexes and D-dimers. The type of surgery with the most pronounced changes was total hip replacement, followed by hemicolectomy, cholecystectomy, and subtotal thyroid resection. CONCLUSION: The total hip replacement and hemicolectomy groups show similar and strong activation of the procoagulatory and fibrinolytic systems. Much less pronounced are the changes during endoscopic cholecystectomy and subtotal thyroid resection. Maximum activation occurs 1 to 2 hours postoperatively. PMID- 10680103 TI - The pharmacokinetics of morphine and lidocaine in nine severe trauma patients. AB - STUDY OBJECTIVE: To study the pharmacokinetic parameters of morphine and lidocaine after a single intravenous (i.v.) bolus in severe trauma patients. DESIGN: Clinical case study. SETTING: Department of Anesthesiology and Intensive Care of a university hospital. PATIENTS: Nine patients, ages 24 to 91 years (mean 54.4 yrs), admitted to the hospital with severe trauma (Injury Severity Score > 20) were included in the study. INTERVENTIONS: After initial evaluation and stabilization, a single i.v. dose of morphine 0.025 mg/kg and lidocaine 1.5 mg/kg was given separately, and blood samples were drawn for each drug serum concentration. MEASUREMENTS AND MAIN RESULTS: Morphine pharmacokinetics was studied in eight patients, lidocaine pharmacokinetics in seven patients, and both drugs were studied in six patients. Morphine clearance 2.5 to 10 ml/kg/min (6 +/- 2.6, mean +/- SD) and volume of distribution 0.28 to 3.30 L/kg (1.4 +/- 1.0) were found to be lower than values described previously for healthy volunteers (33.5 +/- 9 ml/kg/min and 5.16 +/- 1.40 L/kg, respectively), and are similar to those described in trauma patients (5 +/- 2.9 ml/kg/min and 0.9 +/- 0.2 L/kg, respectively). In contrast, lidocaine clearance 4.5 to 9.4 ml/kg/min (6.7 +/- 1.7) and volume of distribution 0.39 to 1.20 L/kg (0.72 +/- 0.28) were similar to the value described in healthy volunteers (10 ml/kg/min and 1.32 L/kg, respectively). CONCLUSION: Changes in pharmacokinetics of drugs eliminated by the liver may occur in patients with severe trauma. The preserved lidocaine clearance indicates an almost normal hepatic blood flow and suggests that other mechanisms may be involved in the lower morphine clearance. The findings may have applications for the treatment of severe trauma patients and suggest that drug monitoring might be needed in some instances so as to avoid toxicity. PMID- 10680104 TI - Influence of thiopental and propofol on postoperative cognitive recovery in the elderly patient undergoing general anesthesia. AB - STUDY OBJECTIVE: To assess mental and psychomotor recovery following induction of anesthesia with thiopental or propofol in elderly patients undergoing general anesthesia. DESIGN: Randomized, prospective, double-blind study. SETTING: Large referral hospital. PATIENTS: 40 elderly patients ASA physical status I-III (> 65 years) undergoing abdominopelvic surgery with an estimated surgical time of at least 90 minutes. INTERVENTIONS: All patients received combined epidural-general anesthesia. After establishing a T6 sensory blockade, patients were randomized to receive either thiopental or propofol for induction of general anesthesia. The induction drug was slowly titrated until loss of eyelash reflex was noted. Thereafter, all patients received desflurane (2% to 3% end-tidal) and 70% nitrous oxide (N2O) in oxygen for maintenance of general anesthesia. To facilitate tracheal intubation, intravenous alfentanil 10 micrograms/kg and atracurium 0.4 mg/kg were administered. Perioperative analgesia was maintained with epidural bupivacaine. MEASUREMENTS AND MAIN RESULTS: A digit substitution test (DSST) and shape-sorter test, as well as patient-generated 100-mm visual analog score (VAS; 0 = minimal and 100 = maximal) for anxiety, sleepiness, and coordination, were performed during the preanesthetic interview, on postanesthesia care unit admission, and at 15, 45, 90, and 120 minutes thereafter. To induce loss of consciousness, either thiopental 2.5 +/- 1.0 mg/kg or propofol 1.6 +/- 0.6 mg/kg was administered. The mean anesthetic time was 109 +/- 30 minutes and 114 +/- 38 minutes for the thiopental and propofol groups, respectively. Emergence, extubation, and orientation times, as well as time to follow commands, were unaffected by patient randomization. Similarly, the DSST and shape-sorter tests, in addition to the patient-generated VAS for pain, anxiety, and coordination, were similar among groups. However, irrespective of treatment modality, return to baseline digit substitution and shape-sorter scores were significantly delayed (p < 0.01). CONCLUSION: When compared to thiopental, propofol does not facilitate improved cognitive recovery in geriatric patients undergoing prolonged surgery. PMID- 10680105 TI - Effects of pretreatment with cisatracurium, rocuronium, and d-tubocurarine on succinylcholine-induced fasciculations and myalgia: a comparison with placebo. AB - STUDY OBJECTIVE: To evaluate the efficacy of cisatracurium, rocuronium, and d tubocurarine in preventing succinylcholine-induced fasciculations and postoperative myalgia in patients undergoing ambulatory surgery. DESIGN: Randomized, prospective, placebo-controlled trial SETTING: Teaching hospital. SUBJECTS: 80 ASA physical status I and II patients scheduled for elective ambulatory surgery with general anesthesia. INTERVENTION: A standardized balanced anesthetic technique was used for all patients. MEASUREMENTS AND MAIN RESULTS: Patients were randomized to receive cisatracurium 0.01 mg/kg, rocuronium 0.06 mg/kg, d-tubocurarine 0.05 mg/kg, or saline, 3 minutes prior to intravenous (i.v.) succinylcholine 1.5 mg/kg. The intensity of fasciculations and intubating conditions were assessed using a four-point rating scale. In addition, the severity of myalgia was assessed using a four-point rating scale in the postanesthesia care unit and at 24 hours postoperatively. No patient complained of any side effects after the administration of the study drug. Fasciculations were observed less frequently (p < 0.05) in the d-tubocurarine and rocuronium groups compared with the placebo and cisatracurium groups. However, there was no difference between the d-tubocurarine group and the rocuronium group (21% vs. 10%, respectively). Although fasciculations occurred less frequently in the cisatracurium group than in the placebo group (59% vs. 85%, respectively), this difference did not reach statistical significance. There was no difference among the four groups in the intubating conditions or the incidence of postoperative myalgia. CONCLUSION: Pretreatment with rocuronium and d-tubocurarine was superior to cisatracurium in preventing succinylcholine-induced fasciculations. However, pretreatment did not have any effect on the incidence of myalgia after ambulatory surgery. PMID- 10680106 TI - Effects of intraoperative glucose administration on circulating metabolites and nitrogen balance during prolonged surgery. AB - STUDY OBJECTIVES: To compare the effects of intraoperative administration of 2.5% glucose or Ringer's solution on metabolism during prolonged surgery. DESIGN: Prospective, randomized study. SETTING: Teaching hospital. PATIENTS: 20 ASA physical status I and II adults patients scheduled for thoracic or abdominal surgery lasting at least 3 hours. INTERVENTIONS: Patients received Ringer's solution (Group R) or 2.5% glucose solution (Group G) 10 ml.kg-1.h-1 during surgery and 2 ml.kg-1.h-1 during the first two postoperative hours (Ringer's or glucose), then 40 ml.kg-1.day-1 of 5% intravenous (i.v.) glucose postoperatively. MEASUREMENTS AND MAIN RESULTS: Plasma glucose, free fatty acids, ketone bodies, lactate, insulin, glucagon, cortisol, and growth hormone concentrations were determined after an overnight fast (T0), on induction of anesthesia (T1), at the end of surgery (T2), 2 hours thereafter (T3), and on the following morning (T4). Capillary blood glucose was determined every 30 minutes from T1 to T2. Urinary nitrogen and 3-methylhistidine were measured every day for 5 days. There were no differences between groups in demographic data, anesthesia, or surgical procedures. All data were comparable at baseline and on the following morning. In Group R, no patient experienced hypoglycemia, but plasma fatty acids and ketone bodies increased during surgery. In Group G, glycemia rose to very high levels, with a significant increase in insulin during surgery. Other hormones were the same within the two groups. Urinary nitrogen and 3-methylhistidine were similar in both groups. CONCLUSION: The absence of glucose infusion in prolonged surgery did not cause hypoglycemia, and no increase in protein catabolism was observed. PMID- 10680107 TI - Duration and recovery profile of cisatracurium after succinylcholine during propofol or isoflurane anesthesia. AB - STUDY OBJECTIVE: To determine the duration and recovery profile of maintenance doses of cisatracurium besylate following succinylcholine, and during propofol or isoflurane anesthesia. DESIGN: Randomized, open-label study. SETTING: Operating suite of a university-affiliated medical center. PATIENTS: Forty ASA physical status I and II adult patients having elective surgery with general anesthesia lasting longer than 90 minutes. INTERVENTIONS: Following a standardized induction sequence, a baseline electromyogram (EMG) was obtained. An intubating dose of intravenous (i.v.) succinylcholine 1.0 mg/kg was administered. Ventilation was maintained with a face mask until the first twitch (T1) of the evoked train-of four (TOF) reached 10% of control when tracheal intubation was performed. Spontaneous recovery from neuromuscular blockade was allowed to occur until the first twitch returned to 25% of control. Patients then were randomized to receive cisatracurium as follows. Group 1: 0.025 mg/kg [0.5 x 95% effective dose (ED95)]; Group 2: 0.05 mg/kg (ED95); Group 3: 0.05 mg/kg (ED95); and Group 4: 0.1 mg/kg (2 x ED95). Anesthesia for Groups 1 and 2 were maintained with isoflurane 1% to 2%, 66% nitrous oxide (N2O) in oxygen (O2), and in Groups 3 and 4, anesthesia was maintained with propofol 80 to 160 micrograms/kg/min, 66% N2O in O2. The TOF evoked EMG was recorded at 10-second intervals. The time for the evoked EMG to spontaneously return to 25%, 50%, and 75% of the original baseline was recorded. MEASUREMENTS AND MAIN RESULTS: There were 10 patients in each of the four groups. The duration of action of cisatracurium 0.05 mg/kg (ED95) after an intubating dose of succinylcholine is 24.5 +/- 10 minutes and 21.3 +/- 9 minutes during anesthesia maintained with isoflurance and propofol, respectively. Doubling the dose of cisatracurium resulted in approximately twice the duration of action (40.2 +/- 7 min) during propofol anesthesia. Following a dose of cisatracurium 0.025 mg/kg (0.5 x ED95), the T1 of the EMG-evoked response did not decrease below 25% in 7 of 10 patients. CONCLUSION: Following succinylcholine, the duration of action of a single dose of cisatracurium 0.05 mg/kg is 20 to 25 minutes during anesthesia maintained with propofol or isoflurane. The duration and recovery profile of cisatracurium is dose dependent during propofol and isoflurane anesthetics. Cisatracurium 0.025 mg/kg is an inadequate maintenance dose following recovery from succinylcholine and it fails to provide adequate surgical relaxation. PMID- 10680108 TI - Comparison of incidence of gastroesophageal reflux and regurgitation associated with timing of removal of the laryngeal mask airway: on appearance of signs of rejection versus after recovery of consciousness. AB - STUDY OBJECTIVES: To compare the incidence of gastroesophageal reflux and regurgitation associated with laryngeal mask airway (LMA) removal when signs of rejecting the LMA, such as swallowing, struggling, and restlessness, were observed and when the patient could open his or her mouth on command. DESIGN: Randomized clinical trial. SETTING: Operating room and recovery room of a tertiary care referral hospital. PATIENTS: 63 ASA physical status I and II adult patients scheduled for elective orthopedic surgery. INTERVENTIONS: Using a standardized general anesthetic technique, patients were allocated randomly to Group A (n = 34; LMA removed when signs of rejection, such as swallowing, struggling, and restlessness, were observed) or Group B (n = 29; LMA removed when the patient could open his or her mouth on command). MEASUREMENTS AND MAIN RESULTS: To detect gastroesophageal reflux throughout anesthesia, a pH monitoring probe was positioned in the lower esophagus on the day before surgery. To assess regurgitation during emergence, a gelatin capsule of methylene blue (50 mg) was swallowed prior to induction. At the end of anesthesia, episodes of reflux and regurgitation of gastric contents were analyzed/determined by pH below 4 and bluish staining of the pharynx and/or LMA, respectively. Physical events such as bucking, straining, and coughing during the arousal phase were recorded in both groups by an independent observer. The incidence of reflux (pH < 4) from the time of the appearance of rejection signs to LMA removal and the total incidence of reflux in Group B were significantly higher than in Group A (p < 0.05). Staining of the LMA and the pharynx by methylene blue was not observed in patients from either experimental group. The number of physical events in Group B during the arousal phase was significantly increased compared to Group A (p < 0.05). Considering all patients in Group A and Group B, physical events were associated with the occurrence of reflux (p < 0.05). Desaturation (SpO2 < 95%) and clinical evidence of aspiration of gastric contents did not occur in either group. CONCLUSION: Maintenance of the LMA until the patient can open his or her mouth on command increases the incidence of gastroesophageal reflux. PMID- 10680109 TI - Low doses of epidural ketamine or neostigmine, but not midazolam, improve morphine analgesia in epidural terminal cancer pain therapy. AB - STUDY OBJECTIVE: To examine analgesia and adverse effects of combination epidural pain therapy consisting of administration of morphine with either low dose of ketamine, neostigmine, or midazolam in terminal cancer pain patients. DESIGN: Randomized double-blind study. SETTING: Teaching hospital. PATIENTS: 48 terminal cancer patients suffering from chronic pain. INTERVENTIONS: Patients were randomized to one of four groups (n = 12). The concept of visual analog scale (VAS), which consisted of a 10-cm line with 0 equaling "no pain at all" and 10 equaling "the worst possible pain" was introduced. All patients were taking oral amitriptyline 50 mg at bedtime. Pain was initially treated with epidural morphine 2 mg twice daily (12-hr intervals) to maintain the VAS below 4/10. Afterwards, VAS scores > or = 4/10 at any time were treated by adding the epidural study drug (2 ml), which was administered each morning, just after the 2-mg epidural morphine administration. The control group (CG) received 2 mg of epidural morphine (2 ml). The ketamine group (KG) received 0.2 mg/kg epidural ketamine (2 ml). The neostigmine group (NG) received 100 micrograms epidural neostigmine (2 ml). The midazolam group (MG) received 500 micrograms epidural midazolam (2 ml). Patients received the study drugs on a daily basis. MEASUREMENTS AND MAIN RESULTS: Duration of effective analgesia was measured as time from the study drug administration to the first patient's VAS score > or = 4/10 recorded in days. The groups were demographically the same. The VAS pain scores prior to the treatment were also similar among groups. Only the patients in the KG demonstrated lower VAS scores compared to the MG (p = 0.018). Time since the epidural study drug administration until patient complaint of pain VAS > or = 4/10 was higher for both the KG and NG compared to the CG (KG > CG, p = 0.049; NG > CG; p = 0.0163). Only the KG used less epidural morphine compared to the CG during the period of study (25 days) (p = 0.003). CONCLUSION: The association of either low-dose epidural ketamine or neostigmine (but not midazolam) to epidural morphine increased the duration of analgesia in the population studied (gt;20 days) compared to the CG and MG (8 to 10 days) when administered in the early stages of terminal cancer pain therapy, without increasing the incidence of adverse effects. PMID- 10680110 TI - Interaction of an implanted pacemaker with a transesophageal atrial pacemaker: report of a case. AB - We report an interaction of a transesophageal atrial pacemaker (TAP) with a permanently implanted pacemaker in a cardiac patient who had undergone ablative therapy for atrial tachyarrhythmia 5 years earlier. The patient's permanent A-V pacemaker was completely inhibited by the TAP, and there was loss of ventricular contractions and blood pressure. The patient required epicardial A-V pacing to overcome the programmed heart rate of 76 bpm. We describe alternative methods to epicardial pacing. We also recommend close inspection of the chest radiograph, which often can reveal the serial numbers of the implanted pacemaker, as a means of identifying the device's functions and programming. PMID- 10680111 TI - Induction of general anesthesia using propofol for cesarean section of a woman with cerebral palsy. AB - A 45-year-old pregnant woman with cerebral palsy was scheduled for cesarean section at 37 weeks' gestation due to the risk of athetotic reaction. Spinal anesthesia appeared difficult to perform due to maintenance position, and because maternal respiratory depression due to athetotic reaction to mechanical stimulation might cause fetal hypoxia. We therefore selected general anesthesia. Propofol and succinylcholine were intravenously (i.v.) administered for induction, and additional propofol was administered i.v. for hemodynamics stabilization. Neonatal Apgar scores were 8 at one minute and 10 at five minutes. No maternal respiratory depression was observed postoperatively, and a healthy baby was successfully delivered. PMID- 10680112 TI - Noncardiogenic pulmonary edema associated with protamine administration during coronary artery bypass graft surgery. AB - Protamine sulfate is the only agent approved to reverse heparin-induced anticoagulation. As with any other drug, protamine has the potential to cause adverse effects that range from mild hypotension to potentially fatal events, such as noncardiogenic pulmonary edema (NCPE) and catastrophic pulmonary vasoconstriction. We report a case of NCPE after the administration of protamine to a patient undergoing coronary artery bypass graft surgery and discuss the diagnosis and management of this severe adverse event. PMID- 10680113 TI - VisualiSAR: a web-based application for clustering, structure browsing, and structure-activity relationship study. AB - VisualiSAR is a program designed to display chemical structures, find similarities and differences between them, and highlight relationships that might exist. The program integrates cluster analysis for the grouping of structurally related compounds, modal analysis of molecular fingerprints for the sorting and highlighting of chemical features, and a Web-based interface for flexibility and ease of use. VisualiSAR has proved useful for a number of applications including the discernment of structure-activity relationships (SAR) of high-volume screening data, and general structure browsing. This article discusses the design of the tool and illustrates two applications. PMID- 10680114 TI - Computational prediction of the three-dimensional structures for the Caenorhabditis elegans tubulin family. AB - In this article we characterize, from a structural point of view, all 16 members of the tubulin gene family of Caenorhabditis elegans (9 alpha-tubulins, 6 beta tubulins, and 1 gamma-tubulin). We obtained their tertiary structures by computationally modifying the X-ray crystal structure of the pig brain alpha/beta tubulin dimer published by Nogales et al. [Nature (London) 1998;391:199-203]. Our computational protocol involves changing the amino acids (with MIDAS; Jarvis et al., UCSF MIDAS. University of California, San Francisco, 1986) in the 3D structure of pig brain alpha/beta-tubulin dimer followed by geometry optimization with the AMBER force field (Perlman et al., AMBER 4. University of California, San Francisco, 1990). We subsequently analyze and compare the resulting structures in terms of the differences in their secondary and tertiary structures. In addition, we compare the pattern of hydrogen bonds and hydrophobic contacts in the guanosine triphosphate (GTP)-binding site for all members of the tubulin family. Our computational results show that, except for gamma-tubulin, all members of the C. elegans tubulin family have similar secondary and 3D structures and that the change in the pattern of hydrogen bonds in the GTP binding site may be used to assess the relative stability of different alpha/beta tubulin dimers formed by monomers of the tubulin family. PMID- 10680115 TI - The mean hydration of carbohydrates as studied by normalized two-dimensional radial pair distributions. AB - The hydration of carbohydrates plays a key role in many biological processes. Molecular dynamics simulations provide an effective tool for investigating the hydration of complex solutes such as carbohydrates. In this article we devise an algorithm for the calculation of two-dimensional radial pair distributions describing the probability of finding a water molecule in a site defined by two reference atoms. The normalized 2D radial pair distribution is proposed as an effective tool for investigating and comparing localized or ordered water sites around flexible molecules such as carbohydrates when analyzing molecular dynamics simulations and the utility of 2D radial pair distributions is demonstrated using sucrose as an example. In this relatively simple structure, 2D radial pair distributions were able to characterize and quantify the importance of two unique interresidue hydration sites in which a water molecule is forming a bridge between the glycopyranosyl and fructofuranosyl residues. The approach is proposed to be a valuable tool for comparing and understanding the hydration of flexible biomolecules such as carbohydrates. PMID- 10680116 TI - The whey acidic protein family: a new signature motif and three-dimensional structure by comparative modeling. AB - Whey acidic proteins (WAP) from the mouse, rat, rabbit, camel, and pig comprise two "four-disulfide core" domains. From a detailed analysis of all sequences containing this domain, we propose a new PROSITE motif ([KRHGVLN]-X-?PF?-X-[CF] [PQSVLI]-X(9,19)-C-?P?-X-[DN]-X-?N? -[CE]-X(5)-C-C) to accurately identify new four-disulfide core proteins. A consensus model for the WAP proteins is proposed, based on the human mucous proteinase inhibitor crystal structure. This article presents a detailed atomic model for the two-domain porcine WAP sequence by comparative modeling. Surface electrostatic potential calculations indicate that the second domain of the pig WAP model is similar to the functional human mucous proteinase inhibitor domains, whereas the first domain may be nonfunctional. PMID- 10680117 TI - A homology modeling method of an icosahedral viral capsid: inclusion of surrounding protein structures. AB - A methodological development is presented for homology modeling of an icosahedrally symmetric assembly of proteins. In the method, a main-chain structure of an asymmetric unit of a protein assembly is constructed and structure refinement is performed, taking the surrounding symmetry-related proteins into consideration with rotational symmetry boundary conditions. To test the procedure, three models of a poliovirus capsid were constructed with different modeling conditions based on the X-ray structure of a rhinovirus capsid. Model S and model N were constructed with and without considering surrounding proteins, respectively. Model N2 was obtained by refinement in rotational symmetry boundary conditions of the structure of model N. The three models were compared with the X-ray structure of a poliovirus capsid. Root mean square deviations and C alpha distances indicate that model S is the most accurate. Examination of the intermolecular short contacts indicates that model S and model N2 are superior to model N, because they do not make severe intermolecular short contacts. Symmetric intermolecular interactions are important for both the structural fragment search and energy minimization to predict better loop structures. The programs developed in this study are thus valuable in homology modeling of an icosahedral viral capsid. PMID- 10680118 TI - The art of molecular graphics. When left isn't right. PMID- 10680119 TI - High performance computational chemistry using parallel PC clusters: an introduction to Beowulf clusters for chemists. PMID- 10680120 TI - Using MOE to visualize molecular orbitals. PMID- 10680121 TI - Virtual network computing: cross-platform remote display and collaboration software. AB - VNC (Virtual Network Computing) is a computer program written to address the problem of cross-platform remote desktop/application display. VNC uses a client/server model in which an image of the desktop of the server is transmitted to the client and displayed. The client collects mouse and keyboard input from the user and transmits them back to the server. The VNC client and server can run on Windows 95/98/NT, MacOS, and Unix (including Linux) operating systems. VNC is multi-user on Unix machines (any number of servers can be run are unrelated to the primary display of the computer), while it is effectively single-user on Macintosh and Windows machines (only one server can be run, displaying the contents of the primary display of the server). The VNC servers can be configured to allow more than one client to connect at one time, effectively allowing collaboration through the shared desktop. I describe the function of VNC, provide details of installation, describe how it achieves its goal, and evaluate the use of VNC for molecular modelling. VNC is an extremely useful tool for collaboration, instruction, software development, and debugging of graphical programs with remote users. PMID- 10680122 TI - [Effect of obesity on the morphology of the left ventricle]. AB - BACKGROUND: To analyze the relationship between obesity in its different degrees and the left ventricle morphology. PATIENTS AND METHODS: M-mode echocardiography was used to estimate the mass, wall thickness and internal dimension of left ventricle in 48 obese women with different degrees of obesity, defined according to the body mass index. 25 women with normal weight were used as controls. RESULTS: The body mass index was correlated with left ventricular mass, as well as with both the wall thickness of the left ventricle and its diastolic internal dimension. The abnormalities in the heart morphology increased according to the obesity degree, ranging from a 59% in the lesser obesity group up to a 100% in the more obese women. The incidence of the left ventricular hypertrophy determined by echocardiography also increased along with the body mass index, ranging from a 29% in the lesser degree of obesity women up to an 82% in the patients with a body mass index > 35 kg/m2. CONCLUSIONS: Obesity, even in its lowest degrees, shows frequent alterations in the heart morphology. This is related with a left ventricular mass increase and a higher incidence of the left ventricular hypertrophy. The left ventricular mass increase is due to an increase in the left ventricular walls thickness and also to a dilatation of its cavity. PMID- 10680123 TI - [Secular increase of adipose tissue in adolescents from Zaragoza from 1980 to 1995]. AB - BACKGROUND: To quantify the differences between anthropometric measurements obtained nowadays in male and female adolescents and those obtained 15 years ago in a sample of similar characteristics. SUBJECTS AND METHODS: We have studied 658 healthy individuals, 329 males and 329 females from 10.0 to 15.0 years of age, from different socioeconomic levels. Weight, height, arm circumference and left skinfold thickness (biceps, triceps, subscapular and suprailiac) have been measured. We have calculated the body mass index (BMI), density, total body fat, percentage of body fat and the body adipose muscular index (BAMI), which is the ratio between body fat (kg) and non fat mass (kg). The values obtained have been compared with those obtained fifteen years ago, in another sample of 1,465 children with similar characteristics. The research team and the material used were the same in both studies. Mean differences were compared using the unpaired t-test. RESULTS: Weight has increased significantly (p < 0.05), except for males at 12 years and females at 13 and at 14 years. Height has increased significantly at all ages and in males and females (p < 0.05). BMI has only increased significantly at 10 and at 11 years in males, and at 10 years in females. Skinfold thicknesses have also increased significantly, except for biceps in males between ages of 12 and 14 years and biceps and subscapular in females at 14 years. Body fat mass and percentage of body fat have also increased significantly and, in consequence, the density has decreased and BAMI has increased both significantly (p < 0.05). CONCLUSIONS: Secular increase in weight and height during fifteen years has occurred. In general, BMI has not increased but skinfold thicknesses and body fat have increased significantly, this fact demonstrates that there has been increase in the body fat compartment. PMID- 10680124 TI - [Lipid profile of the Spanish population: the DRECE (diet and risk of cardiovascular disease in Spain) study. DRECE study group]. AB - BACKGROUND: In Spain the mortality rate due to cardiovascular disease (CVD) is relatively low compared to that of other developed countries. Until now few epidemiological studies have been performed among the global Spanish population to evaluate a relation between CVD risk factors and the lipid profile that could justify our privileged situation. For, this reason, the DRECE study was designed to know the situation at present in Spain respect to the risk of suffering from CVD in particular, the lipid profile. POPULATION AND METHODS: This study included 4,787 subjects (2,324 males and 2,463 females) with an age ranging from 5-60 years, representative of the total Spanish population with these characteristics during the period from 1992 to 1994. Medical history was made for all participants, who also underwent a physical examination. The following parameters were determined: total cholesterol (TC), triglycerides, high-density lipoproteins cholesterol HDLc, cholesterol transported by low-density lipoproteins, LDLc (estimated by the Friedewald's formula), apolipoprotein AI and apoliprotein B (immunoturbidimetry). RESULTS: The results obtained and expressed in mean (SD) show that, although the population has total cholesterol concentrations (190.1 [42.4] and 192.8 [44.8] mg/dl for females and males, respectively) and LDLc (113.9 [37.9] and 117.5 [38.1] mg/dl for females and males, respectively) with values as high as those found in developed countries, the HDLc concentrations (58.6 [13.2] and 51.5 [13.4] mg/dl for women and men, respectively) are also increased and this could be the reason why the mortality rate in Spain caused by CVD is lower than in other countries. CONCLUSIONS: The finding of high HDLc levels and their antiatherogenic role could justify that, at best in part, the rate mortality in Spain is lower than in other developed countries. PMID- 10680125 TI - [Changes in plasma concentrations of lipids and lipoproteins in students from 1992 to 1996: Cuenca study]. AB - BACKGROUND: The aims of this study were to analyze the evolution of plasma lipid and lipoprotein levels in school children of Cuenca city, Spain, between 1992 and 1996; and to study pubertal changes in 1992 cohort. SUBJECTS AND METHODS: A longitudinal observational study was made in 642 schoolchildren 9-16 years old recruited in three public schools of Cuenca city. In 1992, 307 schoolchildren 9 16 years old were examined. On a second examination made in 1996 were examined children of the cohort of 1992 as well as other children who were 9-11 years old in 1996; and belonged to the same schools were recruited. Socio-demographics variables, weight, height, body mass index, systolic blood pressure, diastolic blood pressure and fasting plasma total cholesterol, LDL-C, HDL-C and triglyceride concentrations were determined (and in addition, the apoproteins A-I and B100 and lipoprotein (a) were determined in the second examination). RESULTS: Between the years 1992 and 1996 there has been a falling in total cholesterol concentrations of schoolchildren of the cohort of 1992 due to a decreasing in LDL C and HDL-C levels, the levels total cholesterol ranged from 183.0 mg/dl to 163.3 mg/dl. Comparing the lipid concentrations of the schoolchildren who were 9 to 11 years old in the cohort of 1992 with the lipid concentrations who were 9 to 11 years old in the cohort of 1992 with the lipid concentrations of 1996 it has been found evidence of an important decreasing of total cholesterol and LDL-C, and a moderate increasing in the HDL-C, whereas triglycerides are stable. CONCLUSIONS: Due to the decreasing of total cholesterol and LDL-C levels and the increasing of HDL-C, the lipid profile in schoolchildren of Cuenca city has improved during the 1992-1996 years. Total cholesterol and HDL-C concentrations of 1992 cohort have been a falling during the puberty. PMID- 10680126 TI - [HIV infection: to eradicate it or control it?]. PMID- 10680127 TI - [Declaration of the financial interests of clinical trial researchers]. PMID- 10680128 TI - [Advances in the treatment of the critically burnt patient]. PMID- 10680129 TI - [Assessment of 2 serologic tests (indirect immunofluorescence and ELISA) for the detection of antimalaria antibodies]. PMID- 10680130 TI - [Antivenom serum against Vipera latasti?]. PMID- 10680132 TI - [PubMed: good, beautiful and affordable?]. PMID- 10680131 TI - [Lung leiomyosarcoma 21 years after hysterectomy for uterine leiomyosarcoma: late metastasis?]. PMID- 10680133 TI - [Influenza: disease risks versus vaccine risks]. PMID- 10680134 TI - [Affection-based medicine, family medicine, and university]. PMID- 10680135 TI - [Cytokines and immunomodulation in blood transfusion: a permanent challenge]. PMID- 10680136 TI - [Recurrent myocardial ischemia after spontaneous abortion secondary to oral ergotamine alkaloids]. PMID- 10680137 TI - [Angioedema-urticaria caused by angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists]. PMID- 10680138 TI - [Scleroderma and pregnancy: obstetrical complications and the impact of pregnancy on the course of the disease]. AB - BACKGROUND: To describe the outcome of the pregnancy in patients with scleroderma. PATIENTS AND METHODS: Patients with scleroderma and control group were included in this retrospective study. Two groups were different in pregnant patients with scleroderma: pregnancy before scleroderma (A1) and pregnancy after scleroderma (A2). The presence of clinical problems during pregnancy and the outcome of scleroderma were collected in a questionnaire. Differences in the frequencies of complications were analyzed using the U Mann-Whitney, the chi square or Fisher's exact test when necessary. RESULTS: The frequency of global fetal complications was increased in patients group, but there was no significantly increased frequency when variables were analyzed independently: number of births, miscarriages, fetal deaths, preterm births and low weight full term babies. There was no increased frequency of renal crisis, hypertension or eclampsia. Differences between diffuse and limited subsets were no observed. Improvement of scleroderma was seen in only 3 patients and worsening of skin thickening was experienced by 2 patients. CONCLUSIONS: The pregnant scleroderma patients are a group with high risk pregnancies and therefore well-supervised pregnancies are necessary. PMID- 10680139 TI - [The diet rich in monounsaturated fat modifies in a beneficial way carbohydrate metabolism and arterial pressure]. AB - OBJECTIVE: Two dietary regimens recommended for the reduction of coronary risk, by way of their effects on lipid profile, are the diet low in saturated fat and a diet rich in monounsaturated fats (MUFA). However the effects of these diets on carbohydrate metabolism in healthy subjects are not well known. The objective of this study was to compare the effect of both diets on various parameters of carbohydrate metabolism. METHODS: 41 healthy young males were submitted to 3 consecutive diets, each for a duration of 4 weeks. The first diet was rich in saturated fat (SAT) (38% fat, 20% saturated). The second was rich in carbohydrates following the recommendations of the NCEP-I (National Cholesterol Education Program type I) (28% fat, 47% carbohydrates). The last one was a diet rich in monounsaturated fatty acids (38% fat, 22% MUFA). At the end of each dietary period, blood pressure (BP) and blood levels of glucose, insulin and free fatty acids were determined. 29 subjects were also submitted to an oral glucose tolerance test (OGTT) at the end of each diet. RESULTS: The SAT diet induced the highest levels of insulin after the OGTT. The consumption of the MUFA diet determined the lowest levels of fasting blood glucose (-0.60 mmol/l [13%], p < 0.0002), insulin (-9 microUl/ml [47%], p < 0.0002) and free fatty acids (-0.11 mmol/l [24%], p = 0.006), compared to the NCEP-I diet. Systolic and diastolic blood pressure were higher in the NCEP-I diet than during the other periods (SBP: +6 mmHg compare with SAT [5%], p = 0.0001; and +5 mmHg compare with MUFA [4%], p = 0.0001; DBP: +20 mmHg compare with MUFA [27%], p = 0.0001) and +6 mmHg compared with SAT [8%], p = 0.0001). CONCLUSION: Of the diets most commonly used for the treatment and prevention of arteriosclerosis, a diet rich in monounsaturated fats is the most beneficial for the healthy population from the point of view of carbohydrate metabolism and blood pressure. PMID- 10680140 TI - [The determination of arterial pressure by the physician or the nurse: its relation to ambulatory pressure and left ventricular mass. The MAPA-Madrid Group. Monitorizacion Ambulatoria de la Presion Arterial (Ambulatory Monitoring of Arterial Pressure)]. AB - BACKGROUND: In the present study we evaluated the influence of the observer's status--physician or nurse--on blood pressure levels and the relationship among clinic blood pressure measurement with ambulatory blood pressure and left ventricle mass. PATIENTS AND METHODS: Cross sectional study performed in seven primary care centers. Participating physicians and nurses were trained for blood pressure measurement prior to the study and subsequently retrained at 3 month intervals during the study. Patients included in the study were 122 subjects with mild to moderate hypertension who underwent the following study protocol: a) measurement of clinic blood pressure by physician and nurse, in an independent fashion, on 3 visits; b) clinic-epidemiologic questionnaire; c) conventional hematological and biochemical study; d) electrocardiogram; e) 24-hour ambulatory blood pressure monitoring, f) M-mode and Doppler echocardiography (only in 58 subjects). RESULTS: Nurse-measured blood pressure levels were higher than those determined by physicians (mean differences: 3.9 [6.7] mmHg in systolic blood pressure and 2.6 [5.4] mmHg in diastolic blood pressure). The blood pressure level differences between the two observers were higher in female patients and subjects with low educational level, independently of the observer's gender. Nurse-measured blood pressure was more closely related to ambulatory blood pressure and left ventricle mass than physician-measured blood pressure. CONCLUSIONS: Nurse-measured blood pressure levels are lower than those determined by physicians and more closely related to ambulatory blood pressure and left ventricle mass than physician-measured blood pressure. These data support that nurses, instead of doctors, should routinely measure blood pressure in primary care centers. PMID- 10680141 TI - [Changes in the lipid profile and thyroid function in adult patients with hypopituitarism after substitutive treatment with growth hormone]. AB - BACKGROUND: Adult growth hormone (GH) deficiency is associated with changes in serum lipid levels that can modify after GH substitution. METHODS: We studied 18 patients with GH deficiency treated with GH for 18 or 24 months. RESULTS: A decrease of total cholesterol, LDL with an increase in HDL without triglycerides changes was observed. T4 levels decreased and T3 increased. CONCLUSIONS: The GH substitution treatment in patients with GH deficiency improves the lipid profile and promotes the T4 to T3 conversion. PMID- 10680142 TI - [Pregnancy and systemic autoimmune diseases]. PMID- 10680143 TI - [The monitoring of BCR-ABL mRNA by reverse transcription PCR in patients undergoing allogeneic bone marrow transplantation for acute lymphoblastic leukemia with a positive Philadelphia chromosome]. AB - BACKGROUND: Allogeneic bone marrow transplantation (BMT) has been performed as one mode of cure-oriented therapy for Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL-Ph' positive). However, the clinical significance of the residual BCR-ABL-positive clones after BMT is still controversial. PATIENTS AND METHODS: The BCR-ABL gene (p210 and p190) was prospectively studied by nested RT-PCR in 8 ALL-Ph' positive patients undergoing BMT. RESULTS: All patients received BMT at the time of clinical remission (CR). However, minimal residual disease (MRD) was detected in 7 of them. MRD detected just before BMT seems to be eradicated by BMT protocol. Four patients remained in CR and did not show BCR-ABL transcripts. Other 4 patients, relapsed, demonstrating MRD, which preceded recurrence by a median time of 6 weeks. Three relapsed patients showed p190 transcript and only one, p210 type. CONCLUSIONS: The RT-PCR assay appears to be a useful test for predicting at high risk of relapse after BMT and may identify patients who might benefit from therapeutic interventions. The finding that the expression of p190 BCR-ABL may carry an especially high risk of relapse suggests a different clinical and biologic behaviour between p190 and p210 BCR-ABL. PMID- 10680144 TI - [Resistance to thyroid hormones]. PMID- 10680145 TI - [The molecular changes in thalassemias in Spain. A review of existing studies]. PMID- 10680146 TI - [Primary hepatic pheochromocytoma]. PMID- 10680148 TI - Management of renal cell carcinoma. AB - Surgical resection remains the cornerstone of management for localized renal cell carcinoma. No effective postsurgical adjuvant therapy has been established for patients with locally advanced disease who are at high risk for recurrence. The effective treatment of metastatic kidney cancer remains a challenge. Immunologic therapy with cytokines, such as interferon-alfa (Intron A, Roferon-A) and interleukin-2 (IL-2 [Proleukin]), benefit relatively small numbers of patients. Preclinical research and clinical investigations aimed at identifying new agents and treatment programs with improved antitumor activity against metastases remain the highest priorities in this refractory disease. New immunologic approaches to the treatment of both advanced and high-risk postsurgical disease are focusing on novel vaccine therapies to target both renal epithelial and vascular antigens. PMID- 10680147 TI - [A neck abscess due to Streptococcus pneumoniae in a man with HIV infection]. PMID- 10680149 TI - Paclitaxel in the treatment of advanced urothelial cancer. AB - Median survival in patients with advanced urothelial carcinoma continues to be approximately 1 year following treatment with traditional cisplatin (Platinol) based regimens, which have substantial toxicity. Thus, research is focusing on newer chemotherapeutic agents and novel combination regimens to improve outcomes and tolerability. Paclitaxel (Taxol) demonstrated one of the highest single-agent response rates (42%) in patients with advanced urothelial tumors, prompting extensive evaluation of combination regimens, including paclitaxel/platinum-based doublets or triplets. In many trials, carboplatin (Paraplatin) has been substituted for cisplatin to produce a more convenient, less toxic regimen. These trials have demonstrated that paclitaxel/platinum-based combinations are similar in efficacy to traditional cisplatin-based regimens and are generally better tolerated. Consequently, paclitaxel-based combinations (e.g., paclitaxel/carboplatin) are now considered alternative treatment options for patients with advanced disease, particularly those who are ineligible for clinical trials or are unable to tolerate standard cisplatin-based regimens. Paclitaxel/ifosfamide (Ifex)/cisplatin is another promising alternative regimen for patients who can tolerate cisplatin-based regimens. An ongoing phase III trial comparing paclitaxel/carboplatin and MVAC (methotrexate, vinblastine, Adriamycin, and cisplatin) should clarify the role of paclitaxel-based combinations. Paclitaxel-based combinations are also under evaluation in the adjuvant setting, and future trials may assess their potential role as neoadjuvant therapy or in combination with radiation therapy. PMID- 10680150 TI - Neurocognitive dysfunction in cancer patients. AB - Many cancer patients experience impairments of neurocognitive function, including memory loss, distractibility, difficulty in performing multiple tasks (multitasking), and a myriad of other symptoms. Patients may also concurrently suffer from mood disturbance and symptoms that compromise their ability to function adequately, including fatigue and pain. The etiologies of these problems are diverse and include the direct effects of cancer within the central nervous system (CNS), indirect effects of certain cancers (e.g., paraneoplastic brain disorders), and both diffuse and highly specific effects of cancer treatments on the brain. In addition to these cancer-related causes, patients may have coexisting neurologic or psychiatric disorders that affect their cognition and mood. Careful assessment of patients complaining of neurocognitive or behavioral problems is essential to providing appropriate interventions and maximizing their ability to carry out usual activities. PMID- 10680151 TI - First phase II results of cisplatin/epinephrine in primary liver cancer. PMID- 10680152 TI - Radiotherapy in the management of plasma cell tumors. AB - Most patients with plasma cell tumors receive radiation therapy at some time during the course of their disease. Plasma cell tumors are radio-responsive, but the systemic nature of the disease in most patients limits the application of localized irradiation. In patients with solitary plasmacytomas (osseous and non osseous), radiation therapy is the primary treatment modality. It provides excellent local control that may translate into a long remission and even cure. Adequate dose and careful anatomic planning are essential. In patients with multiple myeloma, effective palliation of pain can be achieved with relatively small fields and low doses of radiation. Hemibody irradiation has been shown to provide cost-effective palliation but is associated with toxicity and has failed to contribute to a more definitive therapeutic approach. Hemibody irradiation is rarely used today. Total-body irradiation is often employed in conditioning regimens prior to autologous or allogeneic stem-cell transplantation for multiple myeloma. However, the magnitude of its contribution to the efficacy of high-dose programs in multiple myeloma remains to be studied. This article explores the rationale for and various aspects of providing effective radiotherapy in patients with plasma cell tumors. PMID- 10680153 TI - Counseling patients with newly diagnosed prostate cancer. AB - Almost 200,000 men are diagnosed with localized prostate cancer each year. With the burgeoning number of treatment options for this disease, the number of patients seeking second opinions from an increasing variety of medical specialties will continue to increase. In 1999 (and for the foreseeable future), there is no single best treatment for any given patient--only options. Each treatment choice has a variety of advantages, as well as a unique spectrum of complications and side effects, most of which are poorly quantified and difficult to compare among treatments. Much more objective data are available regarding the efficacy and necessity of staging tests for further evaluation of patients after diagnosis. Current information about both staging and treatment alternatives, discussed in this article, should be part of the knowledge base of all physicians treating patients with prostate cancer. PMID- 10680154 TI - Are 'platins' on the way out in regimens for NSCLC? PMID- 10680155 TI - Levels of spinal cord injury and predictors of neurologic recovery. AB - A comprehensive physical examination of the patient with acute spinal cord injury is essential in determining the initial level of injury and is the most accurate method of prognosticating neurologic recovery. Understanding neurologic recovery helps predict ultimate functional capability and needs, and helps evaluate the effectiveness of pharmacologic and therapeutic interventions. PMID- 10680156 TI - The respiratory system in spinal cord injury. AB - Respiratory complications are a leading cause of morbidity and mortality during the acute and chronic phases after spinal cord injury (SCI). This article reviews the respiratory impairments resulting from SCI and discusses a range of management strategies to be considered from the acute care phase through long term follow up. The treatment of specific complications and preventive measures are reviewed. PMID- 10680157 TI - The gastrointestinal system and bowel management following spinal cord injury. AB - Gastrointestinal system changes following spinal cord injury (SCI) are generally less obvious than other body system changes. These alterations in function and the response to management, however, may have profound implications on the social, emotional, and physical well-being of the individual with SCI. This article reviews changes in gastrointestinal function following SCI and discusses current issues related to the management of the neurogenic bowel. PMID- 10680158 TI - Surgical procedures of the bladder after spinal cord injury. AB - Surgery of the neurogenic bladder only should be considered after conservative treatment has failed. The goal of this type of surgery of the neuropathic bladder is to restore social continence, approximate normal bladder function, and preserve renal function. Careful selection of procedure and patients, based on medical status and level of ability, combined with realistic expectations, are necessary to ensure a successful outcome. PMID- 10680159 TI - Neurologic recovery and neurologic decline after spinal cord injury. AB - Physicians caring for patients with spinal cord injury facilitate neurologic recovery by optimizing nutrition and general health, by coordinating active exercise and functional training to enhance the underlying synapse growth, reversal of muscle atrophy, and motor learning, and by controlling interfering spasticity. SCI physicians also must monitor for neurologic decline during initial rehabilitation and later in life, diagnose promptly and accurately such decline, and orchestrate the appropriate intervention. PMID- 10680160 TI - Musculoskeletal conditions after spinal cord injury. AB - The impact of musculoskeletal diseases on the overall function and well-being of a person with spinal cord injury (SCI) cannot be overstated. Even relatively minor musculoskeletal problems can lead to significant secondary disabilities and new societal limitations. This article reviews common musculoskeletal problems and related secondary disabilities associated with SCI. Following an overview on the epidemiology and pathophysiology of each condition, a typical case is presented with a discussion about diagnostic challenges and therapeutic options. PMID- 10680161 TI - Metabolic changes in persons after spinal cord injury. AB - Persons with chronic SCI have several metabolic disturbances. As a consequence of inactivity and the body compositional changes of decreased skeletal muscle with a relative increase in adiposity, a state of insulin resistance and hyperinsulinemia has been demonstrated to exist, associated with abnormalities in oral carbohydrate handling. Elevated plasma insulin levels in persons with SCI probably contribute to the cause of frequent dyslipidemia and hypertension. This constellation of metabolic changes represents an atherogenic pattern of CHD risk factors with many of the distinctive features of a cardiovascular dysmetabolic syndrome that is called syndrome X. Reduction in modifiable risk factors for CHD should decrease the occurrence of catastrophic cardiovascular events. There is evidence to suggest that endogenous anabolic hormone levels are depressed in a proportion of individuals with SCI. Depression of serum testosterone and growth hormone/IGF-I levels may exacerbate the adverse lipid and body compositional changes, reduce exercise tolerance, and have deleterious effects on quality of life. Because of immobilization, individuals with paraplegia have osteoporosis of the pelvis and lower extremities, and those with tetraplegia also have osteoporosis of the upper extremities. In addition, there is evidence to suggest that bone loss progresses with time in persons with chronic SCI. This may be caused by chronic immobilization per se or may be a consequence of adverse hormonal changes, including deficiency of anabolic hormones or deficiency of vitamin D and calcium with secondary hyperparathyroidism. Serum thyroid function abnormalities resembling the euthyroid sick "low T3 syndrome" have been reported in those with acute and chronic spinal cord injury. Depressed serum T3 and elevated rT3 in chronic SCI may be caused by associated illness. Current practice has been hesitant to treat abnormal serum thyroid chemistries associated with nonthyroidal illness. Recognition of metabolic abnormalities in individuals with SCI is vital as a first step in improving clinical care. The application of appropriate interventions to correct or ameliorate these abnormalities promises to improve longevity and quality of life in persons with SCI. PMID- 10680162 TI - Sexual function and infertility following spinal cord injury. AB - Changes in sexual function and fertility frequently occur following spinal cord injury (SCI). This article presents an overview of human sexual response and the changes that occur in that response following SCI. This article addresses the issues of childbearing for women with SCI, erectile function for men with SCI, and the issues of fertility and parenting for men and women with SCI. PMID- 10680163 TI - Pain after spinal cord injury. AB - Evaluation of pain in a person with SCI should commence with a determination of the neurologic level and the completeness of injury. The pain then can be localized to one of three regions: above level, at level, or below level. The regional pain then should be categorized either as nociceptive or neuropathic and, after this, subdivided into a specific subtype. An evidence based treatment plan can be devised depending on the specific subtype, which may include physical measures, pharmacologic treatments, behavioral interventions, surgery, or an eclectic combination program. The treatment plan usually can provide some relief for any of the subtypes, although complete relief often is not possible. PMID- 10680164 TI - Medical and rehabilitation issues in the care of children with spinal cord injury. AB - Rehabilitation of the child with SCI involves setting and changing goals appropriate to the child's age, development, and family expectations. Unlike adults with SCI, who may be very close to expected levels of independence at discharge from inpatient rehabilitation, children often require years of outpatient therapy to achieve optimal outcomes. Children with SCI also are at risk for unique complications, which can alter their function. The rehabilitation community extends beyond the usual hospital based practice to include school and transitional services. The focus of rehabilitation changes across stages of development to initially enhance independence in mobility and self-care skills, and later to promote academic achievement, independent living, and employment. PMID- 10680165 TI - Seating assessment and planning. AB - The primary mobility for persons with spinal cord injury (SCI) is seated. This article addresses ways to maximize the SCI patient's functional mobility with proper prescription of seating. Critical components of seating are defined and goals for seating are identified. A format for a postural evaluation as a foundation for seating intervention is included. An overview of equipment for seating highlights unique features and components. PMID- 10680166 TI - Functional neuromuscular stimulation in spinal cord injury. AB - With recent advances in clinical medicine and biomedical engineering, functional neuromuscular stimulation (FNS) can now be added to the psychiatric armamentarium to decrease the debilitating effects of traumatic spinal cord injury. In this article, the components of FNS systems and their evolution in design are presented. The clinical implications of FNS are discussed with respect to upper and lower extremities and bladder applications, and perspectives on future developments and directions are reviewed. PMID- 10680167 TI - Developing clinical practice guidelines for spinal cord medicine. Lessons learned. AB - This article describes the process used by the Consortium for Spinal Cord Medicine to develop evidence-based clinical practice guidelines for managing and treating individuals with spinal cord injury and provides important information on lessons learned and the potential problems to avoid. Issues to consider during the guideline development process include topic selection and explication, methods for selecting the panel chair and panel members, the writing of recommendations and supporting scientific rationales, peer-reviewing guidelines, and the process for disseminating, implementing, and evaluating guidelines. The applicability, advantages, and disadvantages of available evidence and guideline recommendation grading systems and issues arising from the lack of scientific evidence supporting particular recommendations are also discussed. PMID- 10680168 TI - Phytoalexins from crucifers: synthesis, biosynthesis, and biotransformation. AB - Phytoalexins play a significant role in the defense response of plants. These secondary metabolites, which are synthesized de novo in response to diverse forms of stress, including fungal infection, are part of the plants' chemical and biochemical defense mechanisms. Phytoalexins from crucifers are structurally and biogenetically related, but display significantly different biological activities. Here, we review work reporting the chemical structures, synthesis, biosynthesis and metabolism of cruciferous phytoalexins, as well as their biological activity towards different microorganisms. PMID- 10680169 TI - Purification, stabilization and characterization of tomato fatty acid hydroperoxide lyase. AB - Fatty acid hydroperoxide lyase (HPO-lyase) was purified 300-fold from tomatoes. The enzymatic activity appeared to be very unstable, but addition of Triton X100 and beta-mercaptoethanol to the buffer yielded an active enzyme that could be stored for several months at -80 degrees C. The enzyme was inhibited by desferoxamine mesylate (desferal), 2-methyl-1,2-di-3-pyridyl-1-propanone (metyrapone), nordihydroguaiaretic acid (NDGA), n-propyl gallate and butylated hydroxyanisole, suggesting the involvement of free radicals in the reaction mechanism and the existence of a prosthetic group in the active center. However, no heme group could be demonstrated with the methods commonly used to identify heme groups in proteins. Only 13-hydroperoxides from linoleic acid (13-HPOD) and alpha-linolenic acid (alpha-13-HPOT) were cleaved by the tomato enzyme, with a clear preference for the latter substrate. The pH-optimum was 6.5, and for concentrations lower than 300 microM a typical Michaelis-Menten curve was found with a K(m) of 77 microM. At higher alpha-13-HPOT concentrations inhibition of the enzyme was observed, which could (at least in part) be attributed to 2E hexenal. A curve of the substrate conversion as a function of the enzyme concentration revealed that 1 nkat of enzyme activity converts 0.7 mumol alpha-13 HPOT before inactivation. Headspace analysis showed that tomato HPO-lyase formed hexanal from 13-HPOD and 3Z-hexenal from alpha-13-HPOT. A trace of the latter compound was isomerized to 2E-hexenal. In addition to the aldehydes, 12-oxo-9Z dodecenoic acid was found by GC/MS analysis. To a small extent, isomerization to 12-oxo-10E-dodecenoic acid occurred. PMID- 10680170 TI - Hydroquinone: O-glucosyltransferase from cultivated Rauvolfia cells: enrichment and partial amino acid sequences. AB - Plant cell suspension cultures of Rauvolfia are able to produce a high amount of arbutin by glucosylation of exogenously added hydroquinone. A four step purification procedure using anion exchange, hydrophobic interaction, hydroxyapatite-chromatography and chromatofocusing delivered in a yield of 0.5%, an approximately 390 fold enrichment of the involved glucosyltransferase. SDS PAGE showed a M(r) for the enzyme of 52 kDa. Proteolysis of the pure enzyme with endoproteinase LysC revealed six peptide fragments with 9-23 amino acids which were sequenced. Sequence alignment of the six peptides showed high homologies to glycosyltransferases from other higher plants. PMID- 10680171 TI - Characterization of isoforms of hexose kinases in rice embryo. AB - Hexose kinases in rice embryos have been characterized. Six isoforms were detected: i.e. three glucokinases (GK1-3), two hexokinases (HK1 and HK2) and one fructokinase (FK1). Out of these, GK3, HK1 and HK2 were inhibited by mannoheptulose and glucosamine, known inhibitors of hexokinase activity. These inhibitors are also known to be modulators of sugar sensing processes. The results suggest that GK3, HK1 and HK2 may play a role in sensing the cellular sugar status in the rice embryo. PMID- 10680172 TI - Indoxyl-UDPG-glucosyltransferase from Baphicacanthus cusia. AB - The enzyme catalyzing the transfer of glucose from uridine diphosphate glucose to indoxyl yielding the indoxyl glucoside indican was isolated from Baphicacanthus cusia Bremek (Acanthaceae). The indoxyl-uridine diphosphate glucose (UDPG) glucosyltransferase was purified to homogeneity in six chromatographic steps. The decisive step for the recovery of a homogeneous enzyme was the application of immobilized metal affinity chromatography yielding an 863-fold purified enzyme. From a total of 60 substances tested, in addition to the natural substrate 3-OH indole (indoxyl), only 4-OH-, 5-OH-, 6-OH-, and 7-OH-indole were accepted as substrates by the glucosyltransferase. However, the latter substrates were metabolized to varying extent. The optimum pH of the enzyme was 8.5, the optimum temperature was 30 degrees C and the isoelectric point was pH 6.5. The M(r) of the enzyme was determined to be 60 +/- 2 x 10(3). Indoxyl as substrate yielded a K(m) of 1.2 mM, while a K(m) of 1.7 mM was found for UDPG. PMID- 10680173 TI - Sulfation of naringenin by Cunninghamella elegans. AB - A new flavonoid sulfate, naringenin-7-sulfate, was obtained by fermentation of naringenin using the fungus Cunninghamella elegans NRRL 1392 in 23% yield. Structural elucidation of the metabolite was achieved using EIMS, UV, IR, 1D and 2D NMR spectroscopy beside acid and enzyme hydrolyses. PMID- 10680174 TI - Benzoxazinoids-cyclic hydroxamic acids, lactams and their corresponding glucosides in the genus Aphelandra (Acanthaceae). AB - An improved method of sample preparation and simultaneous HPLC separation was developed that allowed the separation of 2,4-dihydroxy-1,4-benzoxazine-3(4H)-one (DIBOA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazine-3(4H)-one (DIMBOA), 2-hydroxy 1,4-benzoxazine-3(2H)-one (HBOA), 2-hydroxy-7-methoxy-1,4-benzoxazine-3(2H)-one (HMBOA) and their corresponding glucosides as well as the benzoxazolinones BOA and MBOA. The amount and distribution of these compounds was determined in the roots of Aphelandra squarrosa and A. fuscopunctata plants. There is a significant difference in the amount and distribution of this substance class in the two species analyzed. The results are discussed in relation to their function as defence compounds and allelochemicals. PMID- 10680175 TI - Variation of DIMBOA and related compounds content in relation to the age and plant organ in maize. AB - We report the variation of all 1,4-benzoxazin-3-one derivatives content detectable in maize with plant age in roots and aerial parts. Our results show that the concentration of hydroxamic acids, 2,4-dihydroxy-7-methoxy-1,4 benzoxazin-3-one glucoside (DIMBOA-Glc) and its 8-methoxylated analogue (DIM2BOA Glc) is high after seed germination and then decreases with plant age. Nevertheless, these compounds continue to be biosynthesised during 6-10 days after germination. Variation in concentration of N-O-methylated DIMBOA-Glc (HDMBOA-Glc) is similar to the one of hydroxamic acids in aerial parts. On the contrary, in roots, its concentration remains relatively stable with plant age. After 10 days, HDMBOA-Glc becomes the main compound in roots. This compound is also present in higher concentration than hydroxamic acids in the oldest leaf of 20-day-old maize. The presence of four other DIMBOA related compounds in maize plants depends on variety, age and tissue. The role of these compounds in plant resistance to aphids is discussed. PMID- 10680176 TI - Trans-4-aminoproline, a phytotoxic metabolite with herbicidal activity produced by Ascochyta caulina. AB - A phytotoxic metabolite, characterized through NMR techniques and synthetic methods as trans-4-aminoproline, was isolated from the culture filtrates of Ascochyta caulina, a promising mycoherbicide for biological control of Chenopodium album. The metabolite, which shows interesting phytotoxic properties, together with ascaulitoxin (recently characterized as N.2-beta-D-glucoside of the unusual bis-amino acid 2,4,7-triamino-5-hydroxyoctandioc acid) and another unidentified compound, compose an active fraction of A. caulina culture filtrates with promising herbicidal properties. When assayed on leaves of host and non host dicots, including wild and cultivated plants, the trans-4-aminoproline showed a wide range of toxicity, with leaves of C. album being the most sensitive. Other interesting aspects were its inefficacy on several monocots, both cultivated and wild, and its lack of antifungal, antibiotic and zootoxic activities. This is the first report on trans-4-aminoproline as naturally occurring compound and phytotoxic metabolite produced by A. caulina. PMID- 10680177 TI - Phytochelatin homologs induced in hairy roots of horseradish. AB - When exposed to excess heavy metals, plants induce phytochelatins and related peptides (all designated as PCAs). Thus, when hairy roots of horseradish (Armoracia rusticana) were exposed for 3 days to cadmium (1 mM) along with reduced glutathione (2 mM), PCA induction occurred. Moreover, a new family of thiol peptides was detected as well as the previously known PCAs, as revealed by postcolumn-derivatization HPLC. Two were isolated and their structures were identified as (gamma-Glu-Cys)n-Gln (n = 3 and 4) by electrospray ionization-mass spectrometer spectra, this being confirmed by chemical synthesis of the peptides. These new analogs constitute the sixth PCA family identified to date. PMID- 10680178 TI - Sesqui- and diterpenes from the liverwort Gackstroemia decipiens. AB - Six rosanes, 5 beta,11 beta-dihydroxy-ros-15-ene, 5 beta,12 beta-dihydroxy-ros-15 ene, 11 beta-hydroxy-7-oxo-rosa-5,15-diene, 1 alpha,5 beta,11 beta-trihydroxy-7 oxo-ros-15-ene, 5 beta,20-epoxy-20-hydroxy-ros-15-ene and 5 beta,20-epoxy-20 methoxy-ros-15-ene along with the enantiomer of the already reported 11 beta hydroxy-rosa-5,15-diene and the known 5 beta-hydroxy-ros-15-ene have been isolated from the liverwort Gackstroemia decipiens. Furthermore, the sesquiterpenes 3-acetoxy-7,11-dihydroxy-farnesa-1,5,9-triene and 1 beta,10 beta epoxy-nardosin-7,11-diene were identified. Their structures were elucidated by NMR spectroscopy. PMID- 10680179 TI - Antinociceptive substances from Incarvillea delavayi. AB - Antinociceptive activities of an Incarvillea delavayi extract, as well as its constituents, 8-epideoxyloganic acid and delavayine A, were evaluated in the acetic acid induced writhing test in mice. An oral administration of the delavayi extract weakly decreased the number of writhings and stretchings in this test, in a dose-dependent manner. Furthermore, orally administered 8-epideoxyloganic acid showed weak antinociceptive activity, whereas administration by subcutaneous injection did not. However, subcutaneous injection of delavayine A, a novel monoterpene alkaloid, showed a more significant level of antinociceptive activity. PMID- 10680181 TI - Polar glycerolipids of Chlamydomonas moewusii. AB - The fatty acid and polar lipid compositions of the unicellular green alga Chlamydomonas moewusii were characterized. Since this organism is an important plant model for phospholipid-based signal transduction, interest was focused on the lipids phosphatidic acid, phosphatidylinositolphosphate and phosphatidylinositolbisphosphate. A phosphatidylinositol:phosphatidylinositolphosphate: phosphatidylinositolbisphosphate ratio of 100:1.7:1.3 was found. The polyphosphoinositides accounted for 0.8 mol% of the total phospholipids and their fatty acid compositions were similar to that of phosphatidylinositol except for the enrichment of linolenic acid in phosphatidylinositol phosphate. Phosphatidic acid accounted for 0.67 mol% of the phospholipids. Major structural glycerolipids were monogalactosyldiacylglycerol (35 mol%), digalactosyldiacylglycerol (15 mol%), sulfoquinovosyldiacylglycerol (10 mol%), diacylglyceryltrimethylhomoserine (16 mol%), phosphatidylglycerol (9 mol%), phosphatidylethanolamine (8 mol%) and phosphatidylinositol (6 mol%). Relative changes in the total fatty acid compositions found during growth on nutrient-limited medium reflected mainly alterations in the compositions of the chloroplast lipids phosphatidylglycerol and monogalactosyldiacylglycerol. [32P]Pi-incorporation studies revealed that it took 6 days before the amount of label in the major phospholipids was proportional to their abundance. PMID- 10680180 TI - Study of sesquiterpene lactones from Milleria quinqueflora on their anti inflammatory activity using the transcription factor NF-kappa B as molecular target. AB - In Central America aerial parts of the Asteraceae Milleria quinqueflora are used in traditional medicine as a remedy for skin infections. Reinvestigation of this plant afforded thirteen sesquiterpene lactones (Sls), three of them are new. All isolated Sls were studied for their anti-inflammatory activity using the transcription factor NF-kappa B as molecular target. NF-kappa B is involved in the synthesis of inflammatory mediators, such as cytokines and chemokines. NF kappa B DNA binding was inhibited at micromolar concentrations by all Sls. PMID- 10680182 TI - Highly oxygenated sesquiterpenes from the liverwort Trocholejeunea sandvicensis. AB - Four pinguisane type sesquiterpenes were isolated from the liverwort Trocholejeunea scandvicensis, together with three aromatic compounds. The structures of the cited compounds were established on the basis of spectroscopic means. The first two compounds were new sesquiterpenes, while the other compounds were previously isolated from other liverworts and lichen sp., respectively. The stereochemistry for lejeuneapinguisanolide was determined by X-ray analysis, a possible biosynthetic pathway to it was postulated. The other new sesquiterpene is lejeuneapinguisenone. PMID- 10680183 TI - Alkaloids from Ruta montana. AB - Two known and four new quinoline and 4-quinolone type alkaloids were isolated from Ruta montana collected from Rommani (Morocco). The known compounds were 1 methyl-4-methoxy-2-quinolone and evolitrine. The structures of the new compounds were established from 1D and 2D NMR experiments including HMQC, HMBC and MS spectral methods as 2-(nonan-8-one)-(1H)-4-quinolone, 2-(nonan-8-one)-4-methoxy quinoline, 2-(nonan-8-one)-N-methyl-4-quinolone and 2-(decan-9-one)-N-methyl-4 quinolone. PMID- 10680184 TI - Benzophenones of Garcinia pseudoguttifera (Clusiaceae). AB - Four biogenetically related benzophenones have been isolated from the Fijian Garcinia pseudoguttifera. They are: 6-hydroxy-2,4-dimethoxy-3,5-bis(3-methyl-2 butenyl)benzophenone (myrtiaphenone-A); 2,2-dimethyl-8-benzoyl-7-hydroxy-5 methoxy-6-(3-methyl-2-butenyl)benzopy ran (myrtiaphenone-B); 2,6-dihydroxy-4 methoxy-3,5-bis(3-methyl-2-butenyl)benzophenone (vismiaphenone-C) and a new benzophenone, 2,2-dimethyl-8-benzoyl-3,7-dihydroxy-5-methoxy- 6-(3-methyl-2 butenyl)-3,4-dihydrobenzopyran (pseudoguttiaphenone-A). Pseudoguttiaphenone-A could be biogenetically derived from vismiaphenone-C. The major component of G. pseudoguttifera was identified as eupha-8,24-dien-3 beta-ol. PMID- 10680185 TI - Tyrosinase catalysed biphenyl construction from flavan-3-ol substrates. AB - Mushroom tyrosinase catalysed oxidation of three flavan-3-ols, viz. catechin, fisetinidol and mesquitol, was conducted to construct biphenyl bonds. Exposure of the flavan-3-ols to tyrosinase and subsequent trapping of the o-quinone intermediates resulted in the formation of novel flavan-3-ol derivatives, the structures of which were elucidated by mono- and two-dimensional 1H-NMR experiments. Application of the methodology resulted in the improved synthesis of the natural flavan-3-ol dimer, mesquitol-[5-->8]-catechin, previously isolated from Prosopis glandulosa. PMID- 10680186 TI - Lipid composition of the extracellular matrix of Botrytis cinerea germlings. AB - Six simple lipid classes (mono-, di- and tri-acylglycerols, free fatty acids, free fatty alcohols and wax esters) were identified by TLC in the extracellular matrix of Botrytis cinerea germlings and the molecular components of each class were characterized using GC-MS. The relative amounts of fatty acids and fatty alcohols within each lipid class were determined by GC-FID. Over all the lipid classes, the most abundant saturated fatty acids were palmitic (ca. 30%) and stearic acid (ca. 22%). Palmitoleic and oleic acids made up ca. 21% and 24% (respectively) of the free fatty acids, while erucic (ca. 4.1%) and linoleic (ca. 3.6%) acids were the most abundant unsaturated fatty acids in the acylglycerides. The acylglycerides also contained almost 35% long chain fatty acids (C20:0 to C28:0). Six fatty acids were identified which had odd-numbered carbon chain lengths (C15:0, C17:0, C19:0, C21:0, C23:0 and C25:0). Of these, pentacosanoic acid made up almost 14% of the fatty acids in the acylglycerides. Three methyl branched chain fatty acids, namely isopalmitic, isoheptadecanoic and anteisopalmitic, were identified in the ECM, all in small amounts. Of the fatty alcohols identified, only palmityl and stearyl alcohols were found in the free form (ca. 57% and 43%, respectively) but arachidyl alcohol (ca. 47%) and 1 octacosanol (ca. 30%) were the most abundant fatty alcohols found in the wax ester fraction. PMID- 10680187 TI - Two flavonoid glycosides from Chenopodium murale. AB - Two new triglycosides, kaempferol-3-O-[(4-beta-D-apiofuranosyl)-alpha-L- rhamnopyranoside]-7-O-alpha-L-rhamnopyranoside and kaempferol-3-O-[(4-beta-D xylopyranosyl)-alpha-L-rhamnopyranoside]-7-O- alpha-L-rhamnopyranoside were isolated from the methanol extract of Chenopodium murale, together with a known diglycoside, kaempferol-3-O-beta-D-glucopyranoside-7-O-alpha-L-rhamnopyranoside. The characterization of the three compounds was achieved by various spectroscopic methods. PMID- 10680188 TI - Koelpinin-A, B and C--three triterpenoids from Koelpinia linearis. AB - Three new triterpenoids, designated as koelpinin-A, B and C and characterised as 28-nor-lup-12,17-dien-3 beta,16 alpha-diol,3 beta-acetoxy-28-nor-lup-12,17-dien 16 alpha-ol and 28-nor-lup-12,17-dien-3 beta-ol-16-one, respectively, together with 30-nor-lup-3 beta-ol-20-one, taraxeryl acetate and germanicol, were isolated, from the aerial parts of Koelpinia linearis. 13C-NMR shifts were assigned after performing APT and DEPT experiments. PMID- 10680189 TI - Oxidation and epimerization of epigallocatechin in banana fruits. AB - To examine the metabolism of proanthocyanidins in banana fruit, (-) epigallocatechin was treated with the homogenate of the fruit flesh to yield (-) gallocatechin and an oxidation product, 1-(3,4,5-trihydroxyphenyl)-3-(2,4,6 trihydroxyphenyl)-2-hydroxy-1-propan one. The latter product is a stable form of a key intermediate in the oxidative metabolism of flavan-3-ols, and is also related to the biogenesis of A-type proanthocyanidins. In addition, treatment of the reaction mixture with o-phenylenediamine afforded monomeric and dimeric phenazine derivatives generated by condensation with the o-quinone form of the oxidation product. PMID- 10680190 TI - Glucosamine in the treatment of osteoarthritis. AB - Glucosamine sulfate, a constituent of cartilage, is evaluated for the treatment of osteoarthritis. Available data suggest that glucosamine decreases pain and improves function in osteoarthritis. Most of the glucosamine studies have methodological flaws or used parenteral formulations, making their data difficult to extrapolate into clinical practice. Glucosamine sulfate is shown to be as good as ibuprofen for osteoarthritis of the knee. Better designed clinical trials of glucosamine are needed to identify its role in the pharmacotherapy of osteoarthritis. PMID- 10680191 TI - Phyto-anti-inflammatories. A systematic review of randomized, placebo-controlled, double-blind trials. AB - Herbal treatments are often used to treat rheumatic symptoms. This systematic review is aimed at determining the clinical efficacy of this approach. Computer literature searches are carried out to locate all placebo-controlled, double blind, randomized trials in this area. Nineteen studies meet the inclusion criteria. They are heterogeneous in terms of remedies tested, patients treated, and trial methodology applied. Most of the studies suggest that herbal remedies can have symptomatic effects beyond placebo. It is concluded that phyto-anti inflammatories have considerable, albeit under-researched, potential in the symptomatic treatment of rheumatic disorders. PMID- 10680192 TI - The Chinese anti-inflammatory and immunosuppressive herbal remedy Tripterygium wilfordii Hook F. AB - Various preparations of Tripterygium wilfordii Hook F (TwHF) have been used in the treatment of a number of autoimmune and inflammatory diseases since the 1960s. Accumulated data from the clinical trials suggest efficacy of this treatment in a number of rheumatic diseases, including rheumatoid arthritis and systemic lupus erythematosus. Studies on the relationship of the chemical components of TwHF and its immunosuppressive and anti-inflammatory effects suggest that diterpenoid compounds with epoxide groups account for the therapeutic effects of this herbal remedy. This herbal remedy is therefore a unique and powerful alternative therapy for autoimmune and inflammatory diseases. PMID- 10680193 TI - Electromagnetic fields and magnets. Investigational treatment for musculoskeletal disorders. AB - Certain pulsed electromagnetic fields (PEMF) affect the growth of bone and cartilage in vitro, with potential application as an arthritis treatment. PEMF stimulation is already a proven remedy for delayed fractures, with potential clinical application for osteoarthritis, osteonecrosis of bone, osteoporosis, and wound healing. Static magnets may provide temporary pain relief under certain circumstances. In both cases, the available data is limited. The mechanisms underlying the use of PEMF and magnets are discussed. PMID- 10680194 TI - Apheresis. AB - The removal of pathologic humors by various methods is an ancient medical remedy used in the management of diseases whose pathophysiology is poorly understood and whose effective treatment modalities are lacking. The contemporary means for such an approach involves apheresis, which is now possible due to advances in blood banking technologies. Apheresis has been used in most of the major rheumatic diseases, in particular systemic lupus erythematosus and rheumatoid arthritis. Although numerous case reports describe clinical benefits of apheresis in rheumatologic disorders, data from clinical trials are discouraging and suggest a limited role for apheresis in rheumatic disease management. PMID- 10680195 TI - Photopheresis and autoimmune diseases. AB - Although several case reports and case series suggest efficacy for photopheresis in the treatment of autoimmune diseases, few controlled studies have been conducted to test this hypothesis. After a decade of interest, multiple case reports, open trials, and one controlled study, the role of photopheresis in autoimmune disease remains to be established. Controlled multi-center trials in rheumatoid arthritis, SLE, and scleroderma may be costly but are clearly necessary for proper evaluation of this therapy. PMID- 10680196 TI - Manual and manipulation techniques for rheumatic disease. AB - Manipulation is practiced primarily by chiropractors and osteopaths and is one of the most commonly utilized alternative treatments for rheumatic diseases. Low back pain and neck pain are the most frequently treated disorders, but manipulation is also used to treat a broad range of rheumatic diseases. Manipulation has been shown to decrease joint pain and normalize function. The mechanisms of action, however, are not well understood. Current theories propose an imbalance of muscle activity is a source of pain that manipulation can relieve through reflexive actions. Such muscle imbalances would exacerbate rheumatic and arthritic conditions, suggesting that manipulation may be an important therapy that is appropriate for early conservative care as part of a comprehensive treatment program. PMID- 10680197 TI - Chiropractic. AB - The chiropractic is a health care profession that offers and purveys a distinctive treatment act. When reduced to its methodology, the chiropractic is hard pressed to demonstrate effectiveness. But as a treatment act, it has gained wide acceptance. The challenge raised in this discussion relates to the "moral hazard" of including the chiropractic treatment act in more general health insurance policies. PMID- 10680198 TI - The evidence for acupuncture as a treatment for rheumatologic conditions. AB - Individuals with rheumatic disorders, particularly those with more severe, chronic conditions, are likely to be frequent users of complementary and alternative medical therapies. Although large-scale clinical trials have yet to be conducted, there is moderately strong evidence that acupuncture may be effective for treating both osteoarthritis and fibromyalgia. The utility of acupuncture in treating rheumatoid arthritis has not been demonstrated in large, randomized controlled trials. Physicians who treat patients with rheumatic conditions should become knowledgeable about the literature on both the effectiveness of acupuncture for these conditions as well as its potential to cause adverse side effects in particular patient groups. PMID- 10680199 TI - Homeopathy and rheumatic disease. AB - Despite a growing interest in uncovering the basic mechanisms of arthritis, medical treatment remains symptomatic. Current medical treatments do not consistently halt the long-term progression of these diseases, and surgery may still be needed to restore mechanical function in large joints. Patients with rheumatic syndromes often seek alternative therapies, with homeopathy being one of the most frequent. Homeopathy is one of the most frequently used complementary therapies worldwide. PMID- 10680200 TI - Yoga. AB - Yoga is an ancient tradition that has been westernized and often practiced for its proposed health benefits. Traditional texts describe its benefits for many types of arthritis. Two limited studies of yoga in osteoarthritis of the hands and carpal tunnel syndrome show greater improvement in pain than in control groups. Yoga uses stretching and improves strength so that it theoretically should be beneficial for some musculoskeletal problems. Yoga merits further study into its cellular and physiologic effects. PMID- 10680201 TI - Ayurvedic medicine and arthritis. AB - The fundamental principles of Ayurveda are briefly reviewed. The ancient classification of arthritis is described along with the comparisons to the modern system. Though the diagnosis is historical and clinical, it is based on the tridosha hypothesis. The Ayurvedic pathogenesis links arthritis to the gut. Management chiefly consists of diet and lifestyle changes, the panchkarma process, and herbal drugs. The rasayana concept of immunomodulation is introduced. Clinical ethno-validation of the ancient therapy is necessary to meet the modern requirements and set up an interface with modern medicine. PMID- 10680202 TI - Exercise, education, and behavioral modification as alternative therapy for pain and stress in rheumatic disease. AB - Stress and pain mechanisms are complex and share many central nervous system pathways. Both are critical issues for patients with rheumatoid arthritis and other connective tissue diseases. The link between stress and neuroendoimmune function suggests that alternative therapies focusing on improved psychologic and metabolic function could significantly change patients' pain outcomes. Programs using alternative therapies such as tai chi and meditation in combination with traditional medications appear to be beneficial for patients with arthritis. These individuals appear to live better lives and may have better long-term outcomes. PMID- 10680203 TI - Mind-body medicine in rheumatologic disease. AB - Research over the last 20 years in Mind-Body Medicine has made significant contributions to the treatment of rheumatic disease. This approach is based on the concept that patients have the ability to influence their experience of illness through directed modification of their thoughts, emotions, and behaviors. This article finds that Mind-Body treatment results in significant, incremental symptom relief and improvement in disability status and well-being beyond that achieved through routine medical care. There is also evidence that these interventions reduce utilization of health care services, despite continuing progression of disease, a finding that has major economic implications for health policy. PMID- 10680204 TI - Prayer and spirituality. AB - Many patients with arthritis are strongly influenced by religious beliefs and often participate in religious healing activities such as prayer and worship attendance. Scientific studies demonstrate, and most patients confirm, that faith and involvement in religious healing activities can be helpful in preventing and treating illness, recovering from surgery, reducing pain, and improving quality of life. To improve the care of patients, clinicians should develop a patient centered, spiritually sensitive form of medical practice in which religious issues are addressed gently and appropriately with dignity, respect, and integrity. PMID- 10680205 TI - [Tick-borne fever (Ehrlichia phagocytophila infection) on a dairy farm in Friesland]. AB - Disease outbreaks, associated with ixodid ticks, among cows on a dairy farm in Friesland, in the north of the Netherlands, were monitored during the summers of 1996, 1997, and 1998. The most important symptoms were a sudden drop in milk yield, fever, apathy, and problems with locomotion. The diagnosis 'tick-borne fever' was confirmed by finding Ehrlichia phagocytophila inclusion bodies in neutrophils of blood smears from sick animals. Significant economic losses due to the disease were recorded each year. Borrelia burgdorferi, although transmitted by the same tick, did not seem to be involved. Further research on tick-borne fever is of particular relevance in connection with recent E. phagocytophila cases diagnosed in dogs and in a human patient in the Netherlands. PMID- 10680206 TI - [Plasma prolactin concentration and the effect of metergoline in pseudopregnant Afghan hounds]. AB - The effects of metergoline, a 5-hydroxytryptamine (serotinin) antagonist, on the plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and on the clinical symptoms of overt pseudopregnancy were studied. Plasma concentrations of prolactin and progesterone were determined in six Afghan hounds with signs of overt pseudopregnancy for 2-3 weeks and in three Afghan hounds that were not pseudopregnant at the time of blood sampling. In the overtly pseudopregnant bitches the plasma concentrations of prolactin before treatment (35.5 +/- 8.5 micrograms l-1) were significantly higher than the plasma concentrations of prolactin of the three bitches that were not pseudopregnant (6.3 +/- 0.5 micrograms l-1); the latter values were similar to those of non psueodopregnant beagle bitches during the total luteal phase. The six pseudopregnant Afghan hounds were treated for 10 days with the antiserotoninergic drug metergoline. At 2 h after the onset of treatment with metergoline, the mean plasma concentration of prolactin had decreased to 10.8 +/- 2.9 micrograms l-1. The plasma concentrations of prolactin continued to decline to 5.4 +/- 1.0 micrograms l-1 at 4 h and to 1.0 +/- 0.1 microgram l-1 during treatment days 3 10. Signs of pseudopregnancy, such as swelling of the mammary glands and digging, decreased during the treatment period. The treatment was associated with mild behavioural side effects such as whimpering and aggressiveness. These side effects are probably not related to suppression of prolactin but are due to a direct effect on serotoninergic pathways in the brain. It is concluded that high plasma concentrations of prolactin are associated with the development and maintenance of pseudopregnancy. The serotonin antagonist metergoline strongly suppresses plasma concentration of prolactine in pseudopregnant dogs and decreases the clinical signs of pseudopregnancy. PMID- 10680207 TI - Initial assessment of patients with neurologic dysfunction. AB - This article describes a practical approach to the initial assessment of patients with neurologic dysfunction. First, the patient's clinical signs are determined by obtaining a medical history and performing an examination. Based on these results, the disease is localized to a particular region of the nervous system. Analysis of the neuroanatomic diagnosis in conjunction with the history is used to establish a list of differential diagnoses. The clinician then recommends appropriate laboratory tests to confirm or exclude the diagnostic possibilities. By following a logical process of clinical reasoning, the practitioner is often able to deduce a clinical diagnosis and prognosis and offer recommendations for treatment. PMID- 10680208 TI - Diagnosis and management of urinary retention. AB - Failure to empty the urinary bladder completely can be attributed to failure of detrusor contractile function, inappropriate outlet resistance, or both. For each of these mechanisms, anatomic, neurogenic, and end-organ (myogenic or idiopathic) abnormalities are possible. The approach to urinary retention involves systematic consideration of neurogenic, obstructive, and functional causes and requires understanding of the neurophysiology and pharmacology of micturation. PMID- 10680209 TI - Selected disorders of muscle and the neuromuscular junction. AB - A variety of disorders affect the muscles or the neuromuscular junction of dogs and cats, most often causing weakness, exercise intolerance, and muscular pain or atrophy. The myopathies are infectious, immune-mediated, inherited or acquired secondary to systemic disease. Acquired myasthenia gravis is a common disorder of the neuromuscular junction, which clinically resembles a myopathy. Reaching a specific diagnosis is essential to determine optimal therapy and prognosis for each of the commonly recognized disorders. PMID- 10680210 TI - Intervertebral disk disease. AB - Neurologic dysfunction is the most common clinical manifestation of intervertebral disk disease. This article is a comprehensive review of intervertebral disk disease emphasizing clinical features, diagnosis and treatment of cervical and thoracolumbar disk disease. Clinical signs are determined by neuroanatomic localization and severity of spinal cord injury. Myelography is most commonly used for determining the location and extent of the disk protrusion/extrusion; however, computed tomography and magnetic resonance imaging are also common adjunctive and primary diagnostic techniques. Summaries from recent studies will give the clinician an improved understanding on how to confront controversial issues regarding prophylactic and therapeutic management and prognosis. PMID- 10680211 TI - Degenerative lumbosacral stenosis. AB - This article reviews the management of degenerative lumbosacral stenosis. Degenerative lumbosacral stenosis occurs when soft tissue and bony changes, possibly in conjunction with abnormal motion of the lumbosacral joint, impinge on the nerve roots or vasculature of the cauda equina. It occurs most frequently in middle-aged dogs of medium to large breed, especially the German Shepherd dog. Common signs are lumbosacral pain, lameness, pelvic limb weakness and ataxia, and urinary incontinence. Diagnosis is based on clinical features and imaging studies. Decompressive surgery is effective in most patients. PMID- 10680212 TI - Spinal fracture or luxation. AB - Spinal trauma is a common cause of spinal cord dysfunction in dogs and cats. When the spine is injured by exogenous injury, the impact often results in vertebral fracture or luxation. As each injury is unique, treatment guidelines have to be individualized to the animal. This article reviews the clinical management including surgical and nonsurgical treatments for animals with vertebral fracture/luxation. PMID- 10680213 TI - Fibrocartilaginous embolism in dogs. AB - Fibrocartilage embolism originating from the intervertebral disk nucleus pulposus may induce acute spinal cord infarction. The main characteristics of this syndrome (acute, nonprogressive transverse myelopathy) are well known by the clinician. However, the impression that this disease is more commonly encountered in giant breeds of dogs, with intumescence involvement and loss of nociception may have been skewed relative to published data. A review of the vascularization peculiarities of the spinal cord explains the still hypothetical pathogenesis and helps to understand the diversity of the clinical presentation. PMID- 10680214 TI - Diskospondylitis and other vertebral infections. AB - This article discusses infections of the spine, including diskospondylitis, vertebral osteomyelitis, and vertebral physitis. Spinal pain is the most common clinical sign. Plain radiography is usually diagnostic; computed tomography and magnetic resonance imaging are useful when the diagnosis is unclear. Serology for Brucella canis and cultures of blood and urine are important in identifying the cause. Appropriate antibiotic therapy is successful in the majority of cases of bacterial spinal infection. PMID- 10680215 TI - Idiopathic epilepsy in dogs. AB - Idiopathic epilepsy is a chronic condition characterized by recurrent seizures for which there is no identifiable cause. It is the most common neurologic disorder in the dog. This article discusses the diagnostic evaluation and rational treatment of dogs with recurrent seizures. Types of seizures, client education, choice of therapy, use of specific drugs, therapeutic monitoring, and nondrug treatments are reviewed. PMID- 10680216 TI - Emergency management of the head trauma patient. Principles and practice. AB - Management of the severely brain-injured dog or cat can be frustrating, especially considering the lack of proven effective therapies for head trauma patients. A working knowledge of the basic pathophysiology of head trauma and intracranial pressure (ICP) dynamics is essential to the logical treatment of head traumatized patients. Prevention and correction of hypotension and hypoxemia are necessary for preventing progressive increases in ICP. Mannitol is recommended in most cases of severe head trauma, but there is little evidence to support the use of glucocorticoids in acutely brain-injured dogs and cats. The role of surgical intervention for head-traumatized dogs and cats is still uncertain, but may be beneficial in some cases. Aggressive, expedient treatment and attentive patient monitoring are key aspects of successfully managing canine and feline head trauma patients. PMID- 10680217 TI - Vestibular dysfunction. AB - Vestibular dysfunction can be caused by damage to the peripheral or central components of the vestibular system. Signs of vestibular disease include ataxia, head tilt, and abnormal nystagmus. This article reviews the pathophysiology, diagnosis, and management of common vestibular disorders in dogs and cats. PMID- 10680218 TI - Is there any health care reform out there? PMID- 10680219 TI - Initial management of trauma by a trauma team: effect on timeliness of care in a teaching hospital. AB - The objective of this study was to determine if timeliness of care would improve after implementation of the team approach in trauma management in a single teaching hospital. To make this determination, we used a before-and-after retrospective cohort series for a 550-bed teaching and tertiary referral hospital that was not a level 1 trauma center. We included all patients who presented to the Emergency Department and who were admitted to St. Paul's Hospital because of trauma during 2 baseline months (May and November 1987; n = 111) and 2 follow-up months (May and November 1990; n = 142). In 1988, a formal trauma team was developed to coordinate the care of trauma patients who were seen in the Emergency Department. Indications for calling the trauma team were based on the criteria of the American College of Surgeons for triage to a trauma center. We calculated elapsed time from assessment in the Emergency Department to arrival of the trauma surgeon, discharge from the Emergency Department, and arrival of the patient in the operating room (for urgent or emergent surgery). We also determined the Revised Trauma Score, the Injury Severity Score (1985 version), the crude mortality ratio, and the Z statistic (population outcome comparison). After implementation of the trauma team, median elapsed time from initial nursing assessment in the Emergency Department to arrival in the operating Room for blunt trauma patients decreased from 11.33 to 4.82 hours (P = .05), but there were no significant differences in any other measures of timeliness, crude mortality, or adjusted mortality. We conclude that implementation of a trauma team in a teaching hospital is associated with a minimal effect on timeliness of care for admitted trauma patients. PMID- 10680220 TI - Admission patterns of an urban level I trauma center. AB - Because trauma admission and hospitalization patterns have profound effects on the organization and utilization of urban trauma-care systems, the objective of this study was to identify and analyze these patterns. As an example, admissions to an urban Level I trauma center were reviewed. Retrospective review of all 2029 trauma admissions to a Level I trauma center was conducted from 1993 to 1996. The result was that most trauma patients were young (40% < 30 years of age) and male (74%). Mechanisms of injury were motor vehicle accident (36%), fall (27%), gunshot (17%), stab (7%), assault (6%), and swimming or diving accident (3%). Half of the patients were directly admitted from the scene. Injury Severity Score, length of stay, and mortality were 14.1 +/- 0.3, 10.5 +/- 0.3 days, and 5.1%, respectively. Admissions tended to occur more frequently between 4:00 PM and midnight (46%), between Friday and Sunday (52%), and between July and October (41%). The following patterns were identified: admissions per year decreased ( 21%) because of reduced penetrating trauma (-43%, P < .01); pediatric patients (< 15 years) had similar incidence of penetrating trauma as adults (ages 15-45). Length of stay for all mechanisms of injury was not statistically different; most mortalities occurred within the first day (33%, P < .01) or after 6 days (36%, P < .01); early mortality was mainly due to penetrating injury (74%, P < .01), whereas late mortality was related to blunt trauma (92%, P < .01). The conclusion was that admission and demographic patterns were identified, which may be useful in the utilization, modification, and future design of trauma systems. PMID- 10680221 TI - Barriers to control of blood glucose in diabetes mellitus. AB - The purpose of this study was to characterize the barriers to tight control of diabetes mellitus. The study collected data from multiple sources at a primary health care clinic in an academic teaching hospital serving an urban population, including patients' charts, structured interviews with patients, a survey of physicians' general perspectives and beliefs concerning diabetes mellitus, and a physician's structured review of barriers to tight control for individual patients. One hundred thirty-five patients with scheduled appointments were eligible for the study, of whom 94% had had a recent hemoglobin A1C (HbA1C). Seventy-seven of 88 patients (88%) who presented for their appointments consented to the interview, 48 of 50 providers (96%) returned useable surveys, and providers completed individual assessments for 96 patients (71%). Patients had a mean age of 61 years, an average of 7.60 diagnoses, and an average of 8.96 prescriptions. Their diabetes control was less than ideal, with less than 15% having normal or near normal control and almost a quarter having poor control. Correlations of HbA1C with age and show rate were seen. Physicians' assessments showed that motivation and understanding of diabetes and compliance with diet and medications correlated with diabetes control. Neither patient knowledge nor physician knowledge appeared to be a problem, nor did either correlate with diabetes control. The number of barriers to control were many, with over half of the patients having five or more barriers. Tight control of blood glucose is felt to be an important aspect of quality care for diabetic patients. In this study, a representative sample of diabetic patients had less than ideal diabetes control. This population was receiving their care in an urban setting and had many comorbid illnesses and many barriers to control. Age, motivation, understanding of the disease, show rate, and compliance with diet and medications had statistical correlations with diabetes control. In order to improve the quality of care for diabetic patients, barriers to care must be addressed. PMID- 10680222 TI - Differences in outpatient corticosteroid prescribing patterns between attending and house staff physicians as an indicator of the quality of supervision. AB - Computerized information systems have become an indispensable source of quality improvement data in the healthcare field. The degree to which we are successful in using these systems is limited only by our ability to ask the right questions. In this study, computerized patient records were used to evaluate the uniformity in the prescribing patterns for oral corticosteroids among house staff and attending physicians as a measure of the adequacy of resident supervision in the outpatient setting. Retrospective analysis of the records of 771 outpatients receiving prescriptions for oral corticosteroid preparations over 1 year in a large tertiary-care university-affiliated Department of Veterans Affairs Medical Center indicated different prescribing patterns for attending physicians and house staff. Additionally, it was noted that house staff tended to manage more complex patients than did attending physicians. We further evaluated the clinical outcomes of these patients to assess the quality, appropriateness, and comparability of care within cohorts of patients and to determine the degree to which resident supervision may have affected outcomes. The study results suggest that there is an opportunity to improve the management of patients treated with oral corticosteroid therapy by increasing staff physician involvement either through direct care of the most complex cases or through enhanced resident supervision. PMID- 10680223 TI - Commentary: health care reform and specialists: the results are not very positive! AB - The present study explored the effects of the current round of health care reforms on practicing physicians in terms of income, cost of care, and improvements in the quality of care. Six neurologists from various parts of the country, all facing widely differing environmental factors, were interviewed. The results demonstrate that health care reform has forced physicians to work more hours to maintain their incomes, has done little to control costs, and has generated little interest in quality improvement plans. In fact, due to referral patterns, quality levels may have decreased under capitated payment plans. The conclusion is that health care must be reformed at the local level and involve physicians with a wide array of specialties. PMID- 10680224 TI - [Saint Liborius, patron of European urology. Iconography found in Croatia and Spain]. AB - The discovery made by the group of Dr. Stella Fatovic-Ferencic in Varazdin (Croatia) of a painting of St. Liborius, patron saint invoked for calculi, and the painting of the Sevillian school (circa 1700) discovered by another group in Spain have led to the retrieval of part of the history of European Urology forgotten 250 years ago. Saint Liborius, bishop of Le Mans (France), died in 397, at the time the barbarian hordes were ravaging the Roman Empire, which had been divided into a Western and an Eastern Empire on the death of Theodosius I. Learning more about St. Liborius is of interest. Here is an example of the Graeco Roman culture of antiguity that is passed on to the present time. The significance of both paintings and their differences are described. The article concludes that on the basis of his biography, St. Liborius should be considered the patron saint of Urology. PMID- 10680225 TI - [Innervation of the human adrenal gland]. AB - OBJECTIVE: To study the innervation of the human adrenal gland by immunohistochemical techniques using monoclonal antibodies against the 200-kDa phosphorylated neurofilament and S-100 protein, specific markers of the axons and Schwann cells, respectively. METHODS: 24 specimens of normal adrenal gland from patients that had undergone radical nephrectomy for a renal tumor were analyzed. The tissue was embedded in paraffin and prepared for analysis by immunohistochemical techniques with the indirect peroxidase method using primary monoclonal antibodies against the 200-kDA phosphorylated neurofilament and protein S-100. RESULTS: The nerves localized in the adrenal gland form a subcapsular plexus where branches arise that extend to the medulla and come into contact with the enterochromaffin cells and medullary neurons. The adrenal neurons are restricted to the medulla; they are dispersed or form ganglia. The satellite cells of the medullary ganglia and the sustentacular cells were positive for S-100. CONCLUSIONS: Most of the nerve fibers penetrate into the human adrenal gland, extending directly to the medulla apparently without coming into contact with the cells of the adrenal cortex and form similar connections to the synaptics with the chromaffin cells and the medullary neurons. The microganglia of the human adrenal gland are dispersed and comprised of few neurons. PMID- 10680226 TI - [Etiology and treatment of penoscrotal skin defects]. AB - OBJECTIVE: The influence of the type and extent of debridement on survival of patients with Fournier's gangrene and the efficacy of reconstruction with grafts and residual skin are analyzed. METHODS: The study comprised 43 patients; 35 Fournier's gangrene, 7 trauma and one tumor. The patients with gangrene were treated by debridement, drainage, amputation and antibiotics. Cutaneous grafts and residual skin were used to repair the defect in a second stage procedure. No testicular avulsion was found in the trauma cases. The penile and scrotal injuries were sutured. One burial and one local flap were performed in two penoscrotal avulsions. Skin from the penis was used to repair partial scrotal defects and grafts were applied on the penis. RESULTS: Mortality (21%) was high in partially debrided Fournier's gangrene, penis amputation and in cases undergoing only drainage. Skin grafts for total loss of penoscrotal cover and penile skin flaps for partial scrotal defects achieved good results. Tension suture was unfavorable in the penis, but was well-tolerated in the scrotum. Burial and local skin flaps were not good solutions. Poor results were obtained by conserving residual skin and grafting only the defect. CONCLUSIONS: Early, wide and repeated debridement procedures reduced the mortality in patients with gangrene, and the best results of reconstruction were obtained with split thickness skin grafts for total loss of penoscrotal cover and penile skin flaps for partial defects of the scrotum. PMID- 10680227 TI - [Renal angiomyolipoma. Ultrasonography and computerized tomography findings]. AB - OBJECTIVE: To analyze the US and CT findings and their value in the diagnosis and follow-up of renal angiomyolipoma. METHODS/RESULTS: The clinical features, US and CT findings in 17 cases of renal angiomyolipoma were reviewed. Patient mean age was 49.1 years, 15 were asymptomatic and two presented with spontaneous bleeding. Two patients with Bourneville's disease presented bilateral lesions. The presumptive diagnosis was based on the US findings in 15 patients and on the CT findings in 16 patients. The remaining patient was evaluated by MRI. Three patients underwent partial nephrectomy and three other patients required total nephrectomy. CONCLUSIONS: Ultrasound is useful in the diagnosis of renal angiomyolipoma and it is probably the method of choice for follow-up. Two thirds were echogenic, although an echogenic lesion is not necessarily fat or viceversa. It is therefore necessary to perform a CT evaluation to make the diagnosis, especially if the tumor produces symptoms. In this series, detection of fat on CT evaluation was diagnostic of renal angiomyolipoma, although fatty tissue can also be found in other tumors or inflammatory lesions. Occasionally, the densitometric findings may not be conclusive due to the artefact of partial volume. The use of 5 mm slices without contrast and volumetric acquisitions for reconstruction can enhance the spatial resolution. It must also be taken into account that the fat content of a lesion can be scanty and may be undetected or not distinguished from other tumors. PMID- 10680228 TI - [Development of a clinical pathway for radical prostatectomy]. AB - OBJECTIVE: To describe the clinical care path for retropubic radical prostatectomy of the La Paz teaching hospital and the results achieved after the first 6 months. METHODS: We have developed a clinical care path for radical prostatectomy with a hospital stay of 6 days. Thirty-one patients submitted to retropubic radical prostatectomy from June to November 1998 were included in the program. The mean length of total, preoperative and postoperative stay were analyzed and compared with those of 31 patients who had undergone radical prostatectomy before the program was developed. Readmissions, adverse effects and patient satisfaction were also analyzed. RESULTS: Of the 31 patients included in the clinical care path, 22 (71%) had a stay equal to or less than the program's length of stay. The mean total, pre and postoperative stay for the group of patients included in the clinical care path were 6.0 days (SD = 1.1), 1 day (SD = 0.0) and 4.9 days (SD = 1.1), respectively. The length of stay was significantly longer before the program was developed [mean total 10.2 days (SD = 4.9), mean preoperative 2.6 days (SD = 2.6) and mean postoperative 7.6 days (SD = 3.6)] (p < 0.001). Twenty-four patients (77.4%) completed the questionnaire on patient satisfaction, which was highly positive, the overall patient satisfaction rate being higher than the 90% standard. There were no readmissions or significant events ascribable to the program. CONCLUSIONS: In our experience, the clinical care path for radical prostatectomy is a useful tool to reduce the unwanted variability. Its design is based on the best possible evidence, therefore the scientific and technical quality, patient satisfaction and efficiency are enhanced. In our view, our results are attainable and feasible in any health care setting. PMID- 10680229 TI - [Expression of protein p53 in superficial transitional carcinoma of the bladder with differing course]. AB - OBJECTIVE: To determine the prognostic value of p53 protein expression in relation to progression of superficial bladder cancer. METHODS: A retrospective study was conducted in which p53 protein was determined in TUR fragments of 18 patients with superficial transitional cell carcinoma of the bladder with no evidence of tumor progression in the last 6 years and in 13 patients with superficial tumors that had become invasive. DO-7 monoclonal antibody was utilized (+if stained nuclei were more than 25%). RESULTS: Expression of p53 protein was found in 9 patients (50%) with bladder tumors that had not progressed and in 6 patients (46.1%) with bladder tumors that had become invasive (p = 0.83). CONCLUSION: Determination of p53 protein was not related with cancer progression in this series. PMID- 10680230 TI - [Clinical and ultrasonographic characteristics of prostatic cancer diagnosed with transrectal biopsy]. AB - OBJECTIVE: To analyze the clinical, ultrasound and pathological characteristics of patients with prostate cancer diagnosed by ultrasound-guided transrectal biopsy who consulted for lower urinary tract symptoms compatible with benign hyperplasia of the prostate. METHODS: From 1994 to 1998 ultrasound-guided transrectal biopsy of the prostate was performed in 1,548 patients aged 49-90 years (mean age 70 +/- 7). Of these, 613 (40%) were diagnosed with cancer. Biopsy was performed in 161 patients (60%) with elevated PSA but no suspicion of cancer on digital rectal examination, and 452 patients with a suspicious DRE. Only 10 of these 452 patients (2%) had a PSA value within the normal ranges. Ninety-seven percent of the patients with cancer were diagnosed at the first biopsy. RESULTS: The DRE findings were compatible with locally advanced cancer in 25% of the patients. An echogenic nodule was detected in 79%; the nodule was hypoechoic in 93% of the cases. Ninety-four percent of the patients with a suspicious DRE versus 37% of those with a normal DRE showed a nodule on ultrasound (p < 0.001). According to the ultrasound and DRE findings, 41% of the patients had a localized cancer. Cancer was diagnosed in 70% of the patients with a nodule; 51% of the cases had high grade carcinoma (score 8, 9 or 10). Sixty percent of the patients with a suspicious DRE had high grade cancer versus 30% of the patients with a normal DRE (p < 0.001). Twenty-three percent of the patients underwent radical prostatectomy. Only 34% of the patients had a localized tumor. CONCLUSIONS: Most of the tumors of the prostate diagnosed in patients who presented with lower urinary tract symptoms compatible with BHP were high grade and locally advanced and therefore not susceptible to cure. PSA determination and digital rectal examination at the primary care level can be useful in detecting prostate cancer in the early stages without significantly increasing costs. PMID- 10680231 TI - [Evidence-based medicine. Usefulness of isolated cystomanometry for the diagnosis of periurethral detrusor-sphincter dyssynergia in patients with suprasacral lesion]. AB - OBJECTIVE: To determine the utility of cystomanometry alone in the diagnosis of periurethral detrusor-sphincter dyssynergia in patients with suprasacral spinal cord lesion. METHODS: Cystomanometry and selective electromyography were performed to evaluate the periurethral sphincter of 40 patients (16 female and 24 male; mean age 46.6 years) with suprasacral spinal cord injury diagnosed by neurological evaluation. RESULTS: The pre-test probability of having dyssynergia was 0.45. The post-test probability, with cystomanometrically demonstrated hyperreflexia, was 0.58. The likelihood ratio utilizing cystomanometry alone was 1.69. CONCLUSION: Cystomanometry alone is not useful in the diagnosis of periurethral detrusor-sphincter dyssynergia in patients with suprasacral spinal cord injury. Periurethral electromyography should be used in combination with the foregoing technique. PMID- 10680232 TI - [Intestinal obstruction secondary to peritoneal carcinomatosis after iatrogenic perforation during TUR for transitional cell carcinoma of the bladder]. AB - OBJECTIVE: A case of intestinal obstruction following transurethral resection (TUR) of a locally advanced transitional cell carcinoma of the bladder is presented. METHODS/RESULTS: The unique features of the present case are described. Laparotomy was performed and peritoneal carcinomatosis was found. Renal function became impaired and the abdominal condition persisted. The patient died five days postoperatively. CONCLUSIONS: Intestinal obstruction following TUR of transitional cell carcinoma of the bladder is an uncommon complication that is mainly due to peritoneal seeding following iatrogenic perforation of the tumor or the bladder wall during resection. Still, TUR is undeniably useful in the diagnosis, staging and treatment of bladder neoplasm. PMID- 10680233 TI - [Reiter syndrome: atypical complication of treatment with BCG]. AB - OBJECTIVE: To report a case of Reiter's syndrome, an uncommon complication after intravesical BCG therapy. The etiology, clinical features, diagnostic difficulties and treatment of this condition are discussed. METHODS/RESULTS: A 56 year-old patient with superficial bladder carcinoma developed conjunctivitis, urethritis and arthritis of the right knee and wrist after the fourth BCG instillation. Other pathologies were ruled out by the clinical and laboratory findings (synovial fluid, cultures, HLA B-27, RF, ANA). A progressive clinical improvement was observed when BCG instillation was discontinued and treatment with indomethacin was started. CONCLUSION: Reiter's syndrome is an infrequent complication that is not related with the BCG strain utilized. Early diagnosis of this condition improves response to therapy. PMID- 10680234 TI - [Giant renal lithiasis]. AB - OBJECTIVE: To describe a case of giant renal calculus. METHODS/RESULTS: A 57-year old female with a previous history of left renal calculus 30 years earlier, consulted for episodes of gross hematuria. The imaging studies showed a giant, spherical calculus, 9 cm in diameter, in the left renal pelvis with hydronephrosis and remaining renal parenchyma functioning. The patient refused surgery. CONCLUSION: Giant renal calculi may progress slowly, without symptoms and without abolishing renal function. PMID- 10680235 TI - [Surgery of abdominal aortic aneurysm in horseshoe kidney. Report of a case and review of the literature]. AB - OBJECTIVE: A case of coexistent abdominal aortic aneurysm and horseshoe kidney is presented. The diagnostic difficulties and the different treatments are discussed. METHODS/RESULTS: A 55-year-old male patient with aneurysm of the abdominal aorta associated with a horseshoe kidney is described and the literature is reviewed. The diagnosis was made by CT and arteriography. Treatment was by the transperitoneal approach, division of the renal isthmus and placement of an aortoiliac dacron graft. CONCLUSION: Horseshoe kidney associated with abdominal aortic aneurysm requiring surgical management is uncommon, but when it occurs, aortic repair is significantly more difficult. PMID- 10680236 TI - [Giant condyloma of the penis. Clinical case]. AB - OBJECTIVE: To report an uncommon case of giant penile condyloma. METHODS/RESULTS: The patient was treated surgically by denudation and fulguration due to the extent of the penile lesion before treatment with topical podophyllin. The histopathological study was determinant in the diagnosis and the postoperative results were excellent. Papillomavirus infection and B. Lowenstein tumor are discussed. CONCLUSIONS: It is recommended to start treatment with topical podophyllin. If the lesion does not improve, resection and fulguration of the condyloma should be performed. The patient should be followed closely for recurrence. PMID- 10680237 TI - [Skin metastasis from transitional cell carcinoma of the bladder]. AB - OBJECTIVE: To report an additional case of skin metastasis from transitional cell carcinoma of the bladder. The clinical features, distribution, differential diagnosis, treatment and survival are reviewed. METHODS: A 73-year-old patient with stage pT2 pN0 pMx carcinoma of the bladder is described. The imaging studies indicated tumor recurrence, but the patient refused reevaluation by endoscopy one year ago. RESULTS: An erythematous, nodular, indurated lesion extending from the hypogastrium to the proximal region of both lower limbs was found. The biopsy findings were compatible with metastatic spread from a carcinoma. CONCLUSION: Although skin metastasis from bladder carcinoma is rare, the number of cases reported is increasing. Skin metastasis is generally limited to the advanced stages of the disease and is an indication of poor prognosis. PMID- 10680238 TI - [Non-specific granulomatous prostatitis with epithelioid morphology]. AB - OBJECTIVE: To report a case of nospecific granulomatous prostatitis. METHODS: A 57-year-old male presented with urinary symptoms (pollakiuria and dysuria), but no fever. On physical examination, the prostate was enlarged and indurated. Urine cultures were negative and blood PSA was slightly elevated. The patient had no previous history of surgery. RESULTS: The histopathological analysis showed granulomatous prostatitis with prominent epithelioid histiocytes. Abundant calculi were also found. The special techniques were negative for fungi and BAAR. No evidence of neoplasm was found. CONCLUSION: Granulomatous prostatitis can mimick a carcinoma, clinically and even histologically, particularly in those cases with prominent epithelioid histiocytes. PMID- 10680240 TI - A new century for international public health. PMID- 10680239 TI - Conservative surgery in small renal tumors: our experience. AB - OBJECTIVE: We evaluated the effectiveness and safety of the nephron-sparing surgery in the treatment of low stage, easily accessible renal cell carcinoma versus radical nephrectomy, the "gold standard" therapy according to data in the literature. METHODS: From 1988 to 1996, 36 patients (11 women and 25 men; mean age 59.6 years) with a small (< or = 5 cm) solitary renal cell carcinoma and a normal contralateral kidney were submitted to tumor enucleation through a transperitoneal approach. Hot ischemia was performed in 14 cases for an average of 16 minutes. In order to control the extent of surgical resection, in all cases frozen step sections of surgical margins were submitted to histopathological examination. The mean follow-up was 40 months. RESULTS: Complete local resection of the renal cell carcinoma was performed in all patients, with preservation to the furthest extent of the parenchyma not affected by the disease. Renal function remained normal in all cases. Bleeding was easily controlled without clamping the renal artery in 22 cases, while hot renal ischemia was necessary in the remaining 14 cases (mean ischemia time 16 minutes). Average blood loss was 450 cc. Frozen sections of the surgical margins were negative in all cases. Only one case of local recurrence was observed one year after the nephron-sparing operation, which was treated by radical nephrectomy. All patients are alive and cancer-free today. CONCLUSIONS: Elective tumor enucleation for low stage and easily accessible renal cell carcinoma can be performed safely and with a low risk of local recurrence. Definition of the appropriate pre-operative diagnostic approach, maximum tumor size, surgical mini-invasive approach and a longer follow-up are required before this procedure can be widely recommended. PMID- 10680241 TI - WHO and the International Diabetes Federation: regional partners. PMID- 10680242 TI - Incidence and outcome of injury in Ghana: a community-based survey. AB - Injury is an increasingly significant health problem in most low-income countries. However, strategies for preventing injury have not been well addressed. The present study was carried out to measure the incidence and outcome of various mechanisms of injury in Ghana in order to provide data for use in developing priorities for injury prevention efforts. For this purpose, using two stage cluster sampling and household interviews, we surveyed 21,105 persons living in 431 urban and rural sites. During the preceding year, 1609 injuries resulting in one or more days of loss of normal activity were reported. Injury related mortality was slightly higher in the urban (83 per 100,000) than in the rural area (53 per 100,000). However, the burden of disability from nonfatal injuries, as assessed by disability days, was higher in the rural (4697 disability days per 1000 person-years) than in the urban area (2671 days per 1000 person-years). Based on incidence rates and disability times, the major types of injury in the urban area were transport-related injury and falls. In the rural area, agricultural injuries predominated, followed by falls and transport-related injury. In rural and urban areas combined, 73% of motor vehicle-related injuries involved commercial vehicles. In this and other similar developing-country settings, injury prevention efforts should focus on falls and on transport safety in both urban and rural areas, with special attention being paid to commercial vehicles. In rural areas, agricultural injuries contributed the largest burden of morbidity, and should be a priority for prevention efforts. PMID- 10680243 TI - Prevalence of hepatitis C virus antibodies and genotypes in asymptomatic, first time blood donors in Namibia. AB - Reported is the prevalence of hepatitis C virus (HCV) in Namibia as determined using a third-generation enzyme-linked immunosorbent assay (ELISA) on samples of blood collected from all asymptomatic, first-time blood donors between 1 February and 31 July 1997 (n = 1941). The HCV seroprevalence was 0.9% (95% confidence interval (CI): 0.5-1.5%) and no associations were detected between a positive HCV serostatus and the person's sex, region of residence, or previous hepatitis B exposure or hepatitis B carrier status, as determined by hepatitis B surface antigen (HBsAg). The only significant association in a logistic regression model was an increase in HCV positivity with increasing age (P = 0.04). Viral RNA was amplified from 2 out of 18 (11.1%) specimens that were ELISA positive. Genotyping of these specimens, by restriction fragment length polymorphism (RFLP), showed the presence of genotypes 5 and 1a. The positive predictive value of using HBsAg positivity as a surrogate screening marker for HCV in Namibian blood donors was poor (1.6%), with low sensitivity (16.7%) and specificity (89.3%), and detecting only 3 out of 18 serologically HCV-positive specimens. The results of this first study of the prevalence and epidemiology of HCV infection in Namibia suggest that donor blood should be screened for HCV by ELISA in order to prevent the transmission of hepatitis C virus. PMID- 10680244 TI - Assessment of cell culture and polymerase chain reaction procedures for the detection of polioviruses in wastewater. AB - WHO considers that environmental surveillance for wild-type polioviruses is potentially important for surveillance for acute flaccid paralysis as a means of confirming eradication of poliomyelitis. The present study investigated methods for detecting polioviruses in a variety of water environments in South Africa. Most polioviruses were isolated on L20B mouse cells, which, however, were not selective: 16 reoviruses and 8 enteroviruses, apparently animal strains, were also isolated on these cells. Vaccine strains of polioviruses were isolated from surface waters during and shortly after two rounds of mass vaccination of children in an informal settlement where there was no sewerage. The results demonstrated the feasibility of poliovirus surveillance in such settlements. It was also evident that neither poliovirus vaccine strains nor other viruses were likely to interfere significantly with the detection of wild-type polioviruses. Optimal isolation of polioviruses was accomplished by parallel inoculation of L20B mouse cells and at least the PLC/PRF/5 human liver and buffalo green monkey (BGM) kidney cell lines. Analysis of cell cultures using the polymerase chain reaction revealed that 319 test samples contained at least 263 human enteroviruses that failed to produce a cytopathogenic effect. This type of analysis thus significantly increased the sensitivity of enterovirus detection. PMID- 10680245 TI - Status of national diabetes programmes in the Americas. AB - Reported are the responses in the latter half of 1997 of all ministries of health in the Region of the Americas to the Declaration of the Americas on Diabetes, which was adopted by the Directing Council of the Pan American Health Organization (PAHO) in 1996 as a basis for national programme development in diabetes. The short-term targets were the designation of national focal points, the preparation of national estimates of the disease burden, and the development and implementation of national strategies and plans to deal with diabetes. The survey found that most countries recognized diabetes as a significant public health problem. In terms of global relevance, a number of lessons have been learned from this exercise: the role of broadly based participation in gaining recognition at the national health policy level; the wide acceptance of an integrated programme model; the relevance of process-related targets to achieve short-term success; and the critical role of having a designated focal point within the managerial approach. PMID- 10680246 TI - Linking the integrated management of childhood illness (IMCI) and health information system (HIS) classifications: issues and options. AB - Differences in the terms used to classify diseases in the Integrated Management of Childhood Illness (IMCI) guidelines and for health information system (HIS) disease surveillance could easily create confusion among health care workers. If the equivalent terms in the two classifications are not clear to health workers who are following the guidelines, they may have problems in performing the dual activities of case management and disease surveillance. These difficulties could adversely affect an individual's performance as well as the overall effectiveness of the IMCI strategy or HIS surveillance, or both. We interviewed key informants to determine the effect of these differences between the IMCI and HIS classifications on the countries that were implementing the IMCI guidelines. Four general approaches for addressing the problem were identified: translating the IMCI classifications into HIS classifications; changing the HIS list to include the IMCI classifications; using both the IMCI and HIS classification systems at the time of consultations; and doing nothing. No single approach can satisfy the needs of all countries. However, if the short-term or medium-term goal of IMCI planners is to find a solution that will reduce the problem for health workers and is also easy to implement, the approach most likely to succeed is translation of IMCI classifications into HIS classifications. Where feasible, a modification of the health information system to include the IMCI classifications may also be considered. PMID- 10680247 TI - Strategies for safe injections. AB - In 1998, faced with growing international concern, WHO set out an approach for achieving injection safety that encompassed all elements from patients' expectations and doctors' prescribing habits to waste disposal. This article follows that lead and describes the implications of the approach for two injection technologies: sterilizable and disposable. It argues that focusing on any single technology diverts attention from the more fundamental need for health services to develop their own comprehensive strategies for safe injections. National health authorities will only be able to ensure that injections are administered safely if they take an approach that encompasses the whole system, and choose injection technologies that fit their circumstances. PMID- 10680248 TI - Auto-disable syringes for immunization: issues in technology transfer. AB - WHO and its partners recommend the use of auto-disable syringes, "bundled" with the supply of vaccines when donor dollars are used, in all mass immunization campaigns, and also strongly advocate their use in routine immunization programmes. Because of the relatively high price of auto-disable syringes, WHO's Technical Network for Logistics in Health recommends that activities be initiated to encourage the transfer of production technology for these syringes as a means of promoting their use and enhancing access to the technology. The present article examines factors influencing technology transfer, including feasibility, corporate interest, cost, quality assurance, intellectual property considerations, and probable time frames for implementation. Technology transfer activities are likely to be complex and difficult, and may not result in lower prices for syringes. Guidelines are offered on technology transfer initiatives for auto-disable syringes to ensure the quality of the product, the reliability of the supply, and the feasibility of the technology transfer activity itself. PMID- 10680249 TI - Anthropotechnological analysis of industrial accidents in Brazil. AB - The Brazilian Ministry of Labour has been attempting to modify the norms used to analyse industrial accidents in the country. For this purpose, in 1994 it tried to make compulsory use of the causal tree approach to accident analysis, an approach developed in France during the 1970s, without having previously determined whether it is suitable for use under the industrial safety conditions that prevail in most Brazilian firms. In addition, opposition from Brazilian employers has blocked the proposed changes to the norms. The present study employed anthropotechnology to analyse experimental application of the causal tree method to work-related accidents in industrial firms in the region of Botucatu, Sao Paulo. Three work-related accidents were examined in three industrial firms representative of local, national and multinational companies. On the basis of the accidents analysed in this study, the rationale for the use of the causal tree method in Brazil can be summarized for each type of firm as follows: the method is redundant if there is a predominance of the type of risk whose elimination or neutralization requires adoption of conventional industrial safety measures (firm representative of local enterprises); the method is worth while if the company's specific technical risks have already largely been eliminated (firm representative of national enterprises); and the method is particularly appropriate if the firm has a good safety record and the causes of accidents are primarily related to industrial organization and management (multinational enterprise). PMID- 10680250 TI - Humanitarian assistance should include aspects of sexuality. PMID- 10680251 TI - Survey of malaria treatment and deaths. PMID- 10680252 TI - Influenza surveillance and dissemination of information to health professionals and the general public in the province of Quebec. PMID- 10680253 TI - Transfusion-transmitted babesiosis in Ontario: first reported case in Canada. PMID- 10680254 TI - World survey of rabies, 1997. PMID- 10680255 TI - CPHA and the global effort to strengthen responses to HIV/AIDS. PMID- 10680256 TI - Incomes and outcomes. PMID- 10680257 TI - [Social capital, health promotion and population health]. PMID- 10680258 TI - Widening regional inequality in premature mortality rates in Manitoba. AB - OBJECTIVE: To describe regional trends in premature mortality in Manitoba. DESIGN: Comparison of all-cause and cause-specific mortality of persons less than age 75 in 11 Regional Health Authority populations over two time periods: 1985-89 and 1990-94. RESULTS: The provincial premature mortality rate declined over the two time periods (4.00/1,000 to 3.72/1,000). Declines were also observed in 9 of 11 regional populations. Premature mortality increased, however, in the 2 regional populations with the highest mortality rates in the first observation period. CONCLUSION: Declining premature mortality in low mortality populations and rising premature mortality in high mortality populations has resulted in a widening of regional mortality rates in Manitoba. Recent policy initiatives in many provinces, including the devolution of authority for the management and delivery of health services and the implementation of population need-based funding formulas to share health care resources among regional health authorities, if implemented, have the potential to partially mitigate the processes producing these widening regional health inequalities. PMID- 10680259 TI - Trends and variations in perinatal mortality and low birthweight: the contribution of socio-economic factors. AB - OBJECTIVE: To examine trends and regional variations in perinatal mortality and low birthweight (LBW) and regional variations in socio-economic risk factors. METHODS: Population-based study of Central West Region of Ontario with approximately 28,000 births annually during the period 1988-1995 using vital statistics records and Census data. RESULTS AND CONCLUSIONS: There was no significant change in the perinatal mortality rate averaging 9.4 per 1,000 births per year. The LBW rate increased from 49.7 to 54.8 per 1,000, while the prematurity rate increased from 56.1 to 75.8 per 1,000. Significant variation occurred in outcomes among different regions, which was partially explained by socio-economic factors. The increases in LBW and prematurity rate emphasize the need for effective targeted services and programs. In their planning and implementation, regional variations in socio-economic factors, and other factors such as: the availability and utilization of services and barriers to access in services, require further evaluation and consideration. PMID- 10680260 TI - Charitable food assistance: what are food bank users receiving? PMID- 10680261 TI - Geographic origin and risk for congenital infection in a Canadian inner city: findings and implications for policy. AB - This study examines associations between geographic origin and risk for congenital infections, through a chart review of women from the St. James Town area of Toronto delivering at Wellesley Hospital in 1996. Foreign-born women (n = 203) were significantly less likely than Canadian-born women (n = 53) to be HBsAg negative (187/193 vs. 48/48; RR = 0.97, 95% CI 0.94-0.99). There was no significant difference in rubella seronegativity, but rubella immunity was unacceptably low in both groups (less than 90%). A number of rubella non-immune women had delivered previously in Canada. Procedures must be implemented to ensure completion of hepatitis B immunization series in affected newborns, and rubella immunization in seronegative women prior to discharge. As well, updating immunization status most become a routine part of the immigration medical examination. PMID- 10680262 TI - The effect of tuberculosis and tuberculosis contact tracing on school function: an exploratory focus group study. AB - SETTING: Selected schools in East York, an ethnically diverse municipality of 110,000 people within Toronto. OBJECTIVE: To explore school staff's attitudes and beliefs about the nature of tuberculosis and its possible effect on the function and culture of schools. DESIGN: Four focus groups of 6-8 school staff, lasting from 1 to 1.5 hours, were held in the spring of 1997 at four different schools deemed to be at high risk for tuberculosis contact tracing. RESULTS: The study identified the following dominant themes: fear of tuberculosis and its impact on school, lack of knowledge and the need for education concerning tuberculosis, and issues in multiculturalism. CONCLUSION: Tuberculosis was perceived by staff of East York schools to be a source of fear. Lack of accurate and reliable information concerning tuberculosis contributes to this situation. Staff identified age-specific and culturally relevant, educational initiatives as means to reduce this fear. PMID- 10680263 TI - [The importance of dietary sodium: the time has come for a public health intervention]. AB - In recent years, many studies have been published regarding the link between sodium intake and high blood pressure. Canadian, American and WHO Guidelines on the treatment of hypertension all indicate salt reduction as an efficient non pharmacologic recommendation. However, due to the lack of clear and specific Canadian legislation on food labelling, consumers are not able to make informed choices of food products on the basis of salt content. The time has come for public health experts to join this debate. PMID- 10680264 TI - Sources of cigarettes for high school students in two Ontario counties: implications for developing a community response. PMID- 10680265 TI - A telephone-based support program for over-the-counter nicotine patch users. PMID- 10680266 TI - The first reported cluster of food-borne cyclosporiasis in Canada. AB - INTRODUCTION: Prior to 1996, sporadic cases of cyclosporiasis in Canada were most often associated with foreign travel and outbreaks throughout the world were associated with contaminated drinking water. In May 1996, the North York Public Health Department was notified of three laboratory-confirmed cases of cyclosporiasis among persons who attended a luncheon at a religious institution. A ceremonial bath (mikvah) was initially identified as a possible source of exposure to contaminated water. METHODS: Guests of a luncheon were interviewed regarding food, beverage and water exposure. The institution kitchen and water sources were inspected and environmental testing was performed. RESULTS: Eating strawberry flan, decorated with rasberries and blueberries, was associated with developing illness (relative risk = 2.13, p = 0.02). There was no evidence that water exposure was associated with illness. DISCUSSION: This event was the index Canadian cluster of a widespread North American outbreak associated with imported Guatemalan raspberries. The local investigation highlights the role of public health departments in multijurisdictional food-borne outbreaks of emerging pathogens. PMID- 10680267 TI - Infant feeding practices in Ottawa-Carleton: the introduction of solid foods. AB - Infant feeding guidelines regarding the introduction of solid foods are generally not well known in Canada. The guidelines recommend that solid foods be introduced between four to six months of age, depending on the developmental readiness of the infant. In order to understand the underlying factors and patterns which contribute to the introduction of solid foods in infants, data were analyzed from three cross-sectional surveys of parents of six-month-old infants from the Ottawa Carleton region (n = 373, 1988; n = 330, 1992; n = 338, 1996) conducted by the Ottawa-Carleton Health Department. Multivariable analysis showed that mothers who: did not breastfeed, were younger, had lower education, smoked or had partners that smoked, and lacked support after birth, were more likely to introduce solid foods before four months of age. These data support the need for nutrition education programs to increase adherence to the new Nutrition for Healthy Term Infants guidelines. PMID- 10680268 TI - Breastfeeding outcomes of women following uncomplicated birth in Hamilton Wentworth. AB - OBJECTIVE: To examine infant feeding practices up to 8 weeks postpartum in Hamilton-Wentworth. METHODS: A cross-sectional survey of 227 women using a pre discharge, self-administered questionnaire, medical record review and follow-up telephone interview. RESULTS: Breastfeeding initiation rate was 85%. By 6-8 weeks postpartum, 30% of women had stopped breastfeeding; 55% had switched to formula within the first 14 days. Infants who did not receive supplementation in hospital were 2.49 times more likely than infants who received supplementation to breastfeed for at least 6 weeks. Although 54% of mothers who initiated breastfeeding reported receiving formula gift packs, no association was found. CONCLUSIONS: The breastfeeding initiation rate appears to have increased in Hamilton-Wentworth since 1995. However, this study reinforces the need to address early cessation and infant supplementation, and raises concern about violation of the WHO/UNICEF International Code of Marketing of Breastmilk Substitutes through mailing of formula coupons. PMID- 10680269 TI - Newfoundland Panel on Health and Medical Care--adult health survey. AB - An adult health survey was undertaken on the island portion of the Province of Newfoundland as part of the Newfoundland Panel on Health and Medical Care. This study aims to develop models of medical care utilization over a seven-year period, before and after a major restructuring of the provincial health care system. A health survey on a large sample (11,789 individuals) permits both descriptive and explanatory analysis at the regional level. The results of the survey are presented first in a descriptive format, as they were included in reports for policy makers and managers, using administrative aggregations. To study regional differences, descriptive and multivariate analyses were done on an aggregation by an urban dimension, which divides the island into three areas. As expected, there were major differences across regions, and complex interactions on selected variables. The usefulness of health surveys for planning and evaluation of health reform is discussed. PMID- 10680270 TI - The effect of season and weather on suicide rates in the elderly in British Columbia. AB - OBJECTIVES: The authors examined the relationship of suicide in the elderly (65 years and older) to season and weather and compared it to that in the younger population (10-64 years). METHODS: Information on suicides and on weather was obtained for British Columbia for the period 1981 to 1991. The association of suicide with season and weather was assessed using Poisson regression. RESULTS: Whereas younger suicides were associated with season, showing a spring-summer peak, elderly suicides were associated with actual weather. They increased with higher mean daily temperature for the current month (RR = 1.16, 95% CI 1.05-1.28 for each 2.5 degrees C change in mean temperature), and with lower mean daily temperature for the preceding three months (RR = 1.12, 95% CI 1.01-1.23). CONCLUSIONS: Elderly suicide rates appear to be affected by deviations of monthly mean temperature from values expected for that time of year. Increased support by service agencies at times of predicted high risk is suggested. PMID- 10680271 TI - Routine child health care in the emergency department. PMID- 10680272 TI - Is God good for your health? The role of spirituality in medical care. AB - Many studies have found that religious belief and practice have a positive effect on physical and mental health, although the topic needs more research. As religious beliefs may affect both health and health-promoting behavior, physicians should try to understand their patients' beliefs. PMID- 10680273 TI - Are strict vegetarians at risk of vitamin B12 deficiency? PMID- 10680274 TI - Is there an advantage to combination therapy with ACE inhibitors and angiotensin II-receptor blockers? PMID- 10680275 TI - Antiviral agents for treating influenza. AB - The new neuraminidase inhibitors zanamivir and oseltamivir are important additions to the treatment of influenza, being the first class of agents active against both influenza A and influenza B. The decision to use these agents rather than amantadine or rimantadine, which are effective only against influenza A, should be based on the age of the patient, antiviral activity, side effect profile, ease of administration, drug interactions, and cost. All of these agents are effective only when started within 24 to 48 hours of onset of symptoms. To avoid inappropriate use of these agents, treatment should be continued only in patients with a confirmed diagnosis of influenza. Although effective in decreasing symptoms, none of these agents prevent pneumonia or hospitalization secondary to influenza. PMID- 10680276 TI - The spectrum of nonalcoholic fatty liver disease: from steatosis to nonalcoholic steatohepatitis. AB - Nonalcoholic fatty liver disease (NAFL) has been recognized only in the past 20 years. Autopsy studies indicate it is remarkably common, especially among obese persons and patients with type 2 diabetes. Although fatty liver alone is usually benign, an identifiable subset of patients may be at risk of progression to cirrhosis and liver failure. The role of liver biopsy is controversial. No specific, effective therapy as yet exists, although management of weight, lipid levels, and glucose levels is recommended. PMID- 10680277 TI - Psoriasis: a clinical update on diagnosis and new therapies. AB - Psoriasis varies widely in its clinical expression, from a single fingernail pit to widespread disfiguring skin lesions and disabling arthritis. Treatments are divided into five levels, providing a framework for approaching this disease according to severity and recalcitrance to previous treatment. Powerful immunosuppressive drugs are showing some success in treating severe cases. PMID- 10680278 TI - Brain metastases: presentation, evaluation, and management. AB - Brain metastases are a common and devastating consequence of cancer and carry a poor prognosis. Nevertheless, physicians can serve their patients well by suspecting, detecting, and treating them appropriately. PMID- 10680279 TI - Glycoprotein IIb/IIIa inhibitors in acute coronary syndromes. AB - Glycoprotein (GP) IIb/IIIa inhibitors are potent antiplatelet agents and represent an exciting breakthrough in the treatment of acute coronary syndromes. However, their safety and cost-effectiveness require further investigation, and more information on risk stratification is needed to clarify which patients benefit the most from empiric use of these agents. PMID- 10680280 TI - Was it preventable? The comprehensive review of inmate suicide. PMID- 10680281 TI - Befriending mental patients: experience in the Ukraine. PMID- 10680282 TI - I.A.S.P. guidelines for suicide prevention. I.A.S.P. Executive Committee. AB - There are a number of ways in which suicide can be prevented. Broad social issues can be influenced by firm advocacy for change in appropriate areas in different countries, such as by restricting access to specific means of suicide and by enhancing health and social services in general. At the individual level, after the establishment of rapport, there should be screening for the presence of specific mental disorders, which, if present, should be treated vigorously. If medication is indicated, the safest drug should be prescribed, although it is emphasized that even if drugs are utilized, nondrug treatment is important for every suicidal person. The focus of supportive therapy, which can be provided by both the helping professions and volunteer organizations, should be the provision of hope for the future, the enhancement of independence, and the learning of different ways of coping with the inevitable stressors of everyday life. If these guidelines are followed, there is every reason to believe that an impact can be made upon the worldwide problem of suicidal behavior. PMID- 10680283 TI - Incarcerated adolescents' distress and suicidality in relation to parental bonding styles. AB - This study examines the relationships between the bonding style of an incarcerated adolescent with parents and his/her current feelings of self-esteem, hopelessness, and suicidal thoughts and attempts. It also investigates differences between bonding to mother and bonding to father. Some 296 incarcerated adolescents were interviewed using the Parental Bonding Instrument. Significant relationships were found between youths' self-esteem, hoplessness, and suicidal behavior and their bonding style. Youths whose parent(s) had a parental bonding style of affectionless control reported the greatest distress, and youths whose parent(s) had an optimal bonding style reported the least distress. Differences were found between bonding styles with the mother and with the father. Attachment theory may be useful in targeting incarcerated youths who have affectionless control bonding with parent(s) for special interventions since these youths are most at risk for psychosocial problems. PMID- 10680284 TI - Risk assessment and suicide prevention in primary care. AB - General practitioners (GPs) are assumed to occupy an important position in the prevention of suicide through the introduction of risk assessment techniques commonly used in psychiatric practice. Despite this theoretical role for primary care services, it remains unclear how frequently GPs implement risk assessment in patients who may be vulnerable to suicide. To address this, a retrospective survey of probable suicides was conducted within a primary care setting utilizing a questionnaire of GPs who had experienced a patient suicide and was augmented by hospital and coroners' records. 85% of questionnaires were returned and 61 deaths were adjudged as suicides during the year long census period. 75% of suicides were male and 54% were aged under 35.28% were in contact with psychiatric services prior to death, although 60% had some diagnosis of mental disorder. GPs had little knowledge of a patient's life circumstances in up to half of cases. Recording of risk assessment occurred in 38% of subjects, was positively associated with prior psychiatric contact (p = 0.001) but negatively associated with presence of physical illness (p = 0.004), older patient age (p = 0.04), and GPs length in practice (p = 0.05). One GP felt their suicide case was preventable. The low rate of risk assessment and limited knowledge of patient lifestyle point to the need for active engagement of GPs in future suicide prevention strategies and should influence the content of training programs in primary care. PMID- 10680285 TI - The cost of hospital care in the year before and after parasuicide. AB - This study, based in Ireland in the Limerick centre of the WHO/EURO Multicentre Study of Parasuicide, tests the hypothesis that the uptake of hospital services increases significantly following an act of parasuicide. To investigate this, the costs of hospital attendance in the year before and in the year after an act of parasuicide are measured and compared. The sample is comprised of the first 100 individuals who attended an acute general hospital following an act of parasuicide after July 1, 1995. Using a computerized patient record system, every hospital attendance is identified, for each individual, in the 12 months before and after the parasuicide act. This includes every visit to the Emergency Room as well as both general and psychiatric inpatient admissions and outpatient attendances. There was a 50% increase in the uptake of hospital services--32% of the sample attended hospital in the year before compared with 48% in the year after. The total yearly costs for the 100 patients almost doubled from IR 53,652 Pounds (Euro 68,138) to IR 104,454 Pounds (Euro 132,657). Generalizing to the 539 individuals who engaged in parasuicide in the Limerick catchment area, total costs increased from IR 289,184 Pounds (Euro 367,264) to IR 563,007 Pounds (Euro 715,019). This study is an initial step toward the more complex task of estimating to what extent the increased uptake of hospital services is due to the consequences of parasuicide and how much is due to other aspects of the patient's health. PMID- 10680286 TI - Suicidal behavior in the Ukraine, 1988-1998. AB - This report studies the available data concerning suicide rates in the Ukraine and points to the importance of appropriate monitoring of suicides and attempted suicides. It illustrates the necessity of collecting this information and of developing "The Ukrainian National Program on Suicide Prevention." Unfortunately, suicide research and publications about suicide rates were prohibited in the former Soviet Union, so some of the data about suicidal behavior in the Ukraine is incomplete. We used the official suicide death statistics of the Ukraine from the Center of Statistics (Ukrainian Ministry of Health) for the period 1988-1998. The overall rate of suicide in the Ukraine is relatively high. Official statistics in the Ukraine show that there were 29.6 suicides per 100,000 population in 1998. The frequency of completed suicide differs in the various regions of the country, suicides being more frequent in the industrially developed regions and in the rural areas of the country than in the cities. In the western part of the Ukraine the frequency of suicide is relatively low (11.1 per 100,000). Between 1988 and 1997 the suicide rate increased by 57%. In 1998 the suicide rate for women was approximately five times lower than that for men. PMID- 10680287 TI - Evolution of a disease. PMID- 10680288 TI - Aid for International Medicine, Inc. (AIM). Our last ten years. PMID- 10680289 TI - Eye problems in the polar regions. PMID- 10680290 TI - Medical mission to Honduras. December 20, 1998-January 2, 1999. AB - This is the story of a team of medical professionals who responded to an appeal from Church World Services in New York to provide two weeks of primary care for Honduran people in areas devastated by Hurricane Mitch. It was a remarkable experience to see how a small team of changing physicians, nurses, and support personnel could function effectively although they had not known each other, or ever worked together before. Over a period of two weeks with two major holiday weekends and an unusual amount of time spent in travel, the team in seven working days provided primary care to about 500 patients ages 1 day to 82 years with limited medical supplies and no laboratory resources. The patients resided in rural villages, without electricity or running water, which could only be reached over poor rural roads. Full credit should be given to the Honduran CCD which provided support services for the dedicated, compassionate American volunteers, and to the Honduran villagers who patiently waited, often in the rain, to be seen. It is hoped that the medical services were as helpful to the recipients as the experience was to the team participants. There is little doubt that long run efforts of CCD and other non-governmental organizations (NGOs) are needed to stimulate overall improvement in the standard of living in rural villages. Future sustained development will take local leadership to stimulate interest in a better future by spacing children so they can have improved education and healthier living conditions. PMID- 10680291 TI - Medical assistance to the Dominican Republic. By health professionals of the Delaware Army National Guard. PMID- 10680292 TI - The medical missions of Rafael Zaragoza, M.D.: from homeland to hometown. PMID- 10680293 TI - A study of 45,X/46,XX mosaicism in Turner syndrome females: a novel primer pair for the (CAG)n repeat within the androgen receptor gene. AB - This paper describes the procedures developed for the determining of diparental/uniparental origin of X chromosomes in mosaic Turner females (karyotype 45,X/46,XX), and accounts for results of the analysis of chromosomal material from 20 girls with Turner syndrome. An (CAG)n repeat within the androgen receptor (AR) gene was selected as a genetic marker. A novel primer pair for amplification of the (CAG)12-30 repeat was designed. These primers gave an amplification product of 338 bp in length and were following (5'-->3'): agttagggctgggaagggtc and cggctgtgaaggttgctgt. Nineteen of the subjects were heterozygous for the selected marker. In 4 cases there were distinct signals from three alleles. The only Turner female in the study who had been previously ascribed a non-mosaic 45,X karyotype by using cytogenetic techniques, proved to be a cryptic mosaic, displaying two alleles of the genetic marker in the more sensitive molecular assay. These results suggest that in most cases 45,X/46,XX mosaicism in Turner females arises through loss of one of the X chromosomes in some cell lines in originally 46,XX conceptuses, rather than through mitotic non disjunction during early embryogenesis in originally 45,X conceptuses. A high sensitivity of the modified assay based on PCR-amplification of the (CAG)n repeat within AR gene proves its usefulness as a tool for studying mosaicism in Turner syndrome. PMID- 10680294 TI - Temporal and spatial variation of inversion polymorphism in two natural populations of Drosophila buzzatii. AB - The inversion polymorphism of the cactophilic fly Drosophila buzzatii was studied in two natural populations. We assessed the temporal changes and microspatial population structure. We observed a significant increase in the frequency of arrangement 2J at the expense of 2ST in both populations. These gene arrangements appear to affect the life-history of flies differently. Environmental heterogeneity explains the karyotype coexistence in nature. The analysis of population structure showed that differentiation of inversion frequencies among individual breeding sites, the rotting clacodes of Opuntia vulgaris, was highly significant. The karyotypic frequencies did not depart significantly from Hardy Weinberg expectations, neither in individual rots nor in the total population. These results suggest that the observed population structure can be easily accounted by random genetic drift. PMID- 10680295 TI - Development of PCR-based markers for thermosensitive genetic male sterility gene tms3(t) in rice (Oryza sativa L.). AB - Development of simple and reliable PCR-based markers is an important component of marker-aided selection (MAS) activities for agronomically important genes in rice breeding. In order to develop PCR-based markers for a rice thermosensitive genetic male sterility gene tms3(t), located on chromosome 6, the nucleotide sequences of four linked RAPD markers OPF18(2600), OPAC3(640), OPB19(750) and OPM7(550) were used to design and synthesize several pairs of specific primers for PCR amplification of the genomic DNA of both the parents IR32364TGMS (sterile) and IR68 (fertile), involved in mapping this gene. For the RAPD marker OPF 18(2600), two pairs of specific primer pair combination from different positions of the sequence resulted in generation of two codominant STS (Sequence Tagged Sites) markers. In case of markers OPAC3(640), OPB19(750) and OPAA7(550) the first two could generate dominant polymorphism, while the last one could not be successful in PCR amplification. Both the codominant STSs with primer combinations F18F/F18RM and F18FM/F18RM were found to be tightly linked to the tms3(t) gene with a genetic distance of 2.7 cM. The sizes of the different alleles in case of F18F/F18RM, F18FM/F18RM combinations were 2300 bp, 1050 bp, and 1900 bp, 1000 bp respectively. The efficiency of marker-assisted selection for this trait was estimated as 84.6%. Polymorphism survey of 12 elite rice lines, indicated that these PCR-based markers for tms3(t) can now be used in selecting TGMS plants at seeding stage in the segregating populations in environment independent of controlled temperature regime. PMID- 10680296 TI - Increased sister chromatid exchange in the peripheral blood lymphocytes of young women who smoke cigarettes. AB - Rates of sister chromatid exchange in dividing human peripheral blood lymphocytes were determined and compared between smoking and non smoking young women between the ages of 16 and 25. Chromosomes block-stained with Giemsa were also examined for chromosome aberrations. A striking difference in the frequency of sister chromatid exchange was found between young women who smoked and those who did not. Smokers scored a significantly higher, F(1) = 15.99, p = 0.0015, rate of sister chromatid exchange than non smokers. Smokers scored a higher mean of SCEs per cell (12.771, SD 3.53) than non smokers (9.712, SD 2.53). Smokers also scored a higher range of SCEs (4 to 28) as opposed to non smokers (4 to 17). No statistical difference was found between smokers and non smokers for the frequency of chromosome aberrations. The significantly higher frequency of exchange in young smoking women may indicate that initial damage to the DNA in many of these women has probably already occurred, thus causing an increased risk of developing cancer later in life. PMID- 10680297 TI - Expression of the heat-shock protein HSP70 in Drosophila buzzatii lines selected for thermal resistance. AB - The level of HSP70 expression induced by a non-lethal high temperature was examined in lines selected for increased thermal resistance and in corresponding control lines of Drosophila buzzatii, in order to test if selection for high temperature resistance leads to an increased level of HSP70 expression. The lines used were selected for up to 64 generations either as adults or through all larval stages. In adult selection lines, hard selection was implemented every second generation after mild heat hardening. In larval selection lines, larvae were exposed each generation to laboratory "natural" selection. Generally lines selected as adults showed a higher HSP70 expression than did controls, both in third instar larvae and in adults. A strong negative response to selection of HSP70 expression was found in all lines that were selected at cycling temperatures during larval development. The results suggests that a trade off between heat resistance in form of HSP70 expression and fecundity/fertility are responsible for the level of HSP70 expression. The effect of the different methods of selection on HSP70 expression suggests that heat resistance constitutes more than one trait. PMID- 10680298 TI - An IgM monoclonal antibody to Japanese encephalitis virus recognizing a cross reactive epitope on nuclear histones. AB - An IgM class of monoclonal antibody (MAb) raised against 'envelope' (E) glycoprotein of Japanese encephalitis (JE) virus, cross reacted with nuclear histones, in addition to recognizing the viral antigen present in the cytoplasm of infected cells by indirect fluorescent antibody (FA) technique. The experiments on histone depletion by the acid treatment of uninfected PS (porcine kidney) cells, revealed the loss of nuclear immunofluorescence (IF) which was regained after the reconstitution of acid treated cells with histones, prior-to reacting with MAb NHA-2. The IgM MAb recognized specifically the viral antigens expressed on the surface of JE virus infected PS cells by a modified indirect FA. The adsorption of MAb NHA-2 with calf thymus histones (type II-AS) showed a comparative higher drop in the reactivity to JE virus (54.2% reduction) as compared to that against uncomplexed histones (33.3%) by ELISA, thus indicating a higher MAb affinity to the former. In contrast, the adsorption of MAb with chicken RBC nuclei resulted in comparatively more reduction in the reactivity to the uncomplexed histones (52.4% reduction) as against JE virus (37.5%), suggesting that DNA plays some role in modifying and presenting these epitopes. The cross-linkage of epitopes by glutaraldehyde treatment of JE virus antigen and histones showed a 2-fold and higher rise in the MAb reactivity as against those with unfixed or methanol fixed antigens (no cross-linkage), suggesting that the epitope is conformation dependent. Thus, histones seem to share a partial conformational homology with 'E' glycoprotein of JE virus and immune reaction with histones might lead to an autoimmune disorder. PMID- 10680299 TI - Cytotoxicity of non O1, non O139 Vibrios isolated from fresh water bodies in Vellore, south India. AB - The samples of plankton, soil sediment and water from a pond, a lake and a moat respectively in and around Vellore were studied for environmental vibrios. Vibrios were isolated from all these specimens after enrichment in alkaline peptone water and subculture on selective media. Non O1, non O139 Vibrio cholerae, Aeromonas spp. and Plesiomonas spp. were isolated. There were no isolates of V. cholerae serogroup O1 and O139. Representative strains of non O1 and non O139 V. cholerae from environmental sources were tested for toxin production in Chinese hamster ovary (CHO) and Vero cell monolayer in microtitre plates. Thirty-three (91.7%) of the 36 strains tested demonstrated cytopathic effect (CPE) in both cell lines indicating their toxigenicity. PCR done on representative strains of non O1 and non O139 V. cholerae showed that none of the strains were positive for ctx A, tcp A-E and tcp C genes. These results indicate that these non agglutinating environmental vibrios produced cytotoxins other than the cholera toxin. PMID- 10680300 TI - Biotyping of Acinetobacter species isolated from clinical samples. AB - We used the biotyping scheme using carbohydrate substrate utilization test with 14 carbon sources to speciate Acinetobacter isolates from blood and cerebrospinal fluid cultures of patients admitted to the postoperative neurosurgery ICU during January to November 1996. Sixty one patients culture positive for Acinetobacter sp. from blood or cerebrospinal fluid were followed up prospectively. Among these patients, 40 patients had clinically diagnosed infections like bacteriemia or meningitis while in 21 patients the isolation was regarded as contaminants. A. baumanniii was the most common isolate associated with clinical infections while A. lwoffii was more likely to be an environmental contaminant. PMID- 10680301 TI - Changing trend in susceptibility pattern of Streptococcus pneumoniae to penicillin in India. AB - Prior to 1995 all strains of Streptococcus pneumoniae isolated at a tertiary care hospital in south India were uniformly susceptible to penicillin. However, since late 1995 strains of S. pneumoniae with intermediate resistance to penicillin have been observed. Altogether there were 25 such isolates, 9 from invasive (5 from CSF as well as blood, 1 from pleural fluid and 3 from CSF alone) and 16 from noninvasive sites (6 from throat, 6 from sputum, 3 from eye and 1 from ear) respectively, thus 4.6 per cent of S. pneumoniae showed intermediate resistance of a total of 535 strains studied so far. The minimum inhibitory concentration (MIC) values of penicillin, erythromycin, chloramphenicol and cefotaxime were determined by agar dilution method and for confirmation, E test was carried out for penicillin alone. The MIC range obtained for penicillin was between 0.125-1.0 microgram/ml. Kirby-Bauer disc diffusion method was adopted for testing of erythromycin, chloramphenicol, co-trimoxazole, cefotaxime, tetracycline and vancomycin. We observed that none of the strains with intermediate resistance to penicillin were multidrug resistant. These strains belonged predominantly to serotype 14 (n = 10), 7B (n = 9), 19A (n = 3), 7F (n = 2) and 23F (n = 1). Clonality was not observed in the 5 representative strains subjected to Box A finger printing method. PMID- 10680302 TI - Yeast colonisation & fungaemia in preterm neonates in a tertiary care centre. AB - Seventy consecutive preterm neonates who stayed in the hospital for more than seven days between March and October 1996, were studied for colonisation at oral, umbilical, groin, and rectal areas and for fungaemia. Overall, 71.4 per cent of the neonates were colonised and colonisation occurred within 24 h in 38 per cent preterm neonates. Neonates weighing less than 1500 g were colonised more frequently at more than one site and had higher load of yeast. Candida albicans (19%), Pichia (Hansenula) anomala (17.5%), C. tropicalis (13.2%), C. parapsilosis (12.3%) and Trichosporon cutaneum (10.0%) were the predominant colonising yeasts. Fungaemia was detected in 22.8 per cent of preterm neonates with predominance of P. anomala fungaemia (62.5%). Prematurity, male sex, broad spectrum antibiotic therapy, intubation and higher colonising rate were identified as significant risk factors for development of fungaemia. Except one strain of C. tropicalis, all yeast strains were sensitive to commonly used systemic antifungal agents. Study highlights the importance of routine surveillance of yeast colonisation of preterm neonates with identifying possible risk factors. PMID- 10680303 TI - A comparison of esmolol & diltiazem for heart rate control during coronary revascularisation on beating heart. AB - This prospective study was done to compare the control of heart rate and haemodynamics during coronary artery revascularisation without cardiopulmonary bypass using either esmolol or diltiazem. Sixty adult patients with one or two vessel coronary artery disease, were randomly divided into 2 equal groups. Group A received a 500 micrograms/kg loading dose of esmolol followed by a 100 micrograms/kg/h infusion, for control of heart rate during surgical anastomosis of the coronary vessel. While Group B received 0.15 mg/kg diltiazem as a loading dose followed by a 5 mg/h infusion for heart rate control, during the anastomosis. It was seen that heart rate control was better in Group A, 51.4 (+/- 1.3) beats/min, than in Group B, 69.6 (+/- 3.9) beats/min but the decrease in heart rate was significant in both the groups at peak effect compared to respective predrug values. Group A patients had unchanged systemic resistance and pulmonary artery wedge pressure but mean pulmonary artery pressure and pulmonary vascular resistance were significantly raised. Group B patients had decreased systemic resistance, mean pulmonary artery pressure and pulmonary artery wedge pressure, and reduced right ventricular stroke work index. We concluded that although esmolol provided dramatically slower heart rates, during surgery, the resulting elevations in mean pulmonary artery pressure and pulmonary vascular resistance would require caution if used in patients with underlying right ventricular dysfunction from ischaemia or infarction. Diltiazem by virtue of its effects on systemic vascular resistance, cardiac output, and lowering of mean arterial pressure may be a better choice in hypertensive patients. PMID- 10680304 TI - Millennial editorial message. PMID- 10680305 TI - Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin: an application of oxidative stress markers to cancer risk assessment of dioxins. AB - Dioxins are known to be a class of highly toxic and persistent environmental contaminants. Among them the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been the most intensively studied, and it has been classified as a human carcinogenic substance by the International Agency for Research on Cancer (IARC). Although the mechanism of carcinogenesis by TCDD is unclear, it is now considered to have act a cancer promoter. In this review, was discuss the ability of TCDD to induce oxidative stress in vivo, the mechanism of the oxidative stress induction, and how oxidative stress relates to the development of cancer. We then discuss the advantages of measuring the level of oxidative stress in people exposed to dioxins in epidemiological studies for cancer risk assessment. We also discuss several methods of measuring the level of oxidative stress in humans. PMID- 10680306 TI - Characteristics of coronary heart disease in Japanese taxi drivers as determined by coronary angiographic analyses. AB - Several epidemiological studies have shown that the prevalence of ischemic heart disease is higher in occupational drivers than in people with other occupations. Although occupation categories can be surrogate measures for coronary risk factors, the relationships between taxi driving and severity of coronary heart disease (CHD) has not been investigated. Even more important, the contribution of risk factors to the severity of CHD in taxi drivers remains unclear. Our study tested the hypothesis that taxi driving could be associated with the severity of CHD. We also examined the relation between this occupation and risk factors and social lifestyle. We analyzed the coronary angiograms of 57 consecutive male taxi driver patients and compared them with those of 215 age-adjusted male non-taxi driver patients. The number of diseased vessels and risk factors were compared between two groups. The prevalence of myocardial infarction and multi-vessel disease was higher in the taxi-driver patients than in the non-taxi-driver patients. The taxi-driver patients had higher prevalence of body mass index (BMI), diabetes, and smoking, higher levels of low-density lipoprotein cholesterol (LDL-C), and lower levels of apolipoprotein AI (ApoAI). Multiple logistic regression analysis showed that multi-vessel disease was associated with BMI and diabetes mellitus in taxi-driver patients. The taxi-driver patients were characterized by more extensive coronary atherosclerosis associated with higher prevalence of diabetes mellitus and obesity. These characteristics may be explained by in part their working environment. PMID- 10680307 TI - Laboratory evaluation of welder's exposure and efficiency of air duct ventilation for welding work in a confined space. AB - CO2 arc welding in a confined space was simulated in a laboratory by manipulating a welding robot which worked in a small chamber to experimentally evaluate the welder's exposure to welding fumes, ozone and carbon monoxide (CO). The effects of the welding arc on the air temperature rise and oxygen (O2) concentration in the chamber were also investigated. The measuring points for these items were located in the presumed breathing zone of a welder in a confined space. The time averaged concentrations of welding fumes, ozone and CO during the arcing time were 83.55 mg/m3, 0.203 ppm and 0.006%, respectively, at a welding current of 120A-200A. These results suggest serious exposure of a welder who operates in a confined space. Air temperature in the chamber rose remarkably due to the arc heat and the increase in the welding current. No clear decrease in the O2 concentration in the chamber was recognized during this welding operation. A model of air duct ventilation was constructed in the small chamber to investigate the strategy of effective ventilation for hazardous welding contaminants in a confined space. With this model we examined ventilation efficiency with a flow rate of 1.08-1.80 m3/min (ventilation rate for 0.40-0.67 air exchanges per minute) in the chamber, and proved that the exposure level was not drastically reduced during arcing time by this air duct ventilation, but the residual contaminants were rapidly exhausted after the welding operation. PMID- 10680308 TI - Drafts in cold environments--the significance of air temperature and direction. AB - This paper concerns the influence of air temperature and of the direction of drafts on subjective and physiological responses. In three experimental series 58 healthy persons (50 men, 8 women, 20-29 yrs) were exposed to drafts in overall 446 experimental sessions. Drafts were applied either horizontally or vertically with mean air velocities of 0.1 to 0.3 m/s and a turbulence intensity of 50%. Air temperature was varied between 11 and 23 degrees C and metabolic rates between < 70 and 156 W/m2. These parameters were kept constant during the single one-hour sessions. The subjects were dressed for thermal neutrality. Draft-induced annoyance was registered every 5 minutes using a list of prescribed body parts and skin temperature was measured at the forearm and at the neck. Subjective and physiological responses were systematically related to air temperature. Draft induced general annoyance, draft-induced local annoyance (neck, forearm) and the drop of the corresponding skin temperature were inversely related to air temperature. Concerning the direction horizontal drafts seem to cause somewhat stronger reactions. The predictive model developed by Toftum underestimates the percentage of persons annoyed. A modified version increases the predictive power significantly. PMID- 10680309 TI - Evaluation of workers' exposure to dust, ammonia and endotoxin in poultry industries at the province of Isfahan, Iran. AB - This study was conducted to assess various environmental exposure measurements (total dust, ammonia and endotoxin) of poultry workers at the province of Isfahan, Iran. The results show that the workers who worked in enclosed systems of parent stock barns have the highest exposure to total and respirable dust: 21.3 +/- 3.2 and 4.6 +/- 0.9 mg/m3, respectively. In comparison with different ages of chicken, the highest concentration of total and respirable dust were 5.4 +/- 0.7 and 3.3 +/- 0.7 mg/m3 in the 45th day. In the above mentioned situation, the results of endotoxin concentrations were 20.6 +/- 1.1, 23.6 +/- 2.2, 21.3 +/- 1.2 and 26.8 +/- 1.8 ng/m3, respectively. Ammonia concentrations had the highest rate in enclosed systems of laying hens in winter and the 45th day of chicken age, measuring 33.2 +/- 5.2 and 20.2 +/- 3.0 mg/m3, respectively. PMID- 10680310 TI - An electromyographic study of two different types of ballpoint pens- investigation of a one hour writing operation. AB - Recently there has been an increasing incidence of occupational cervicobrachial disorders (OCD) and writer's cramp in office workers using ballpoint pens in writing operations. For the sake of workers who use ballpoint pens, it is essential to prevent such health hazards. It has been observed that a strong gripping pressure on the ballpoint pen significantly contributes to the development of these conditions. The present authors have been developing a new ballpoint pen by altering the grip area in such a way as to reduce the gripping pressure, and thus prevent OCD. The purpose of this study is to compare our ballpoint pen (new pen) with a conventional ballpoint pen (conventional pen) for the load that they exert on the upper limb during one hour of continuous writing. Electromyograms (EMG) and upper limb pain scores are used as indicators. The conventional pen used was selected from commercially available ballpoint pens widely used in offices. The grip area is cylindrical with an 8.3 mm diameter. It is manufactured of hard plastic, which can make it feel rigid and slippery to the user. The new pen has a cylindrical grip area that flares out at the bottom, near the pen-tip, and has a diameter ranging from 11.9-13.6 mm. In addition, the grip is constructed of a 2 to 3 mm-thick silicon rubber sleeve that is softer and less slippery in comparison with the conventional pen. Twelve students (5 males and 7 females) without any preexisting cervicobrachial disorders were asked to transcribe an English text for one hour, alternately using the two kinds of pens. The EMG of the flexor pollicis brevis was measured and recorded every second, while subjective pain scores were recorded every five minutes for the thumb, forefinger, middle finger, thenar, forearm extensor (forearm) and shoulder. The EMG of the flexor pollicis brevis and the pain scores for the thumb, forefinger, middle finger, forearm and shoulder were significantly lower for the new pen than for the conventional pen. These results suggest that after an hour of continuous writing, the new pen reduces the muscle load on the upper limb, and therefore mitigates fatigue in this area. PMID- 10680311 TI - Blood lead levels in copper smelter workers in Japan. AB - Lead exposure of workers in a Japanese copper smelter was assessed by determining lead levels in blood, air and flue cinder at the copper smelting processes. All the samples were analyzed for lead by atomic absorption spectrometry. Mean lead levels of air were highest at the anode department followed by the converter, smelter and blend departments. The mean level of blood lead of the workers in the anode department was also the highest among the four smelting departments. The mean blood lead levels of the workers in each department were positively correlated with their air lead levels (r = 0.99, p < 0.01). This study indicates therefore that workers in copper smelters have been exposed to lead in their workplace. Though this finding has already been reported in preceding studies, the Ordinance on Prevention of Lead Poisoning in Japan has not included copper smelter into its target job categories if their lead concentration in the raw material is less than 3%. The limitation of the present Ordinance which defines the targets by the types of job and not by the actual exposure, is discussed. PMID- 10680312 TI - Sleep problems in white-collar male workers in an electric equipment manufacturing company in Japan. AB - In order to clarify poor sleep habits and prevalence in sleep problems (disturbances) of healthy male white-collar workers and the relationship of these to age and job type, a total of 1,161 Japanese employees of an electric equipment manufacturing company (aged 23 to 59, mean 37 years) were surveyed by means of a mailed questionnaire. The workers were asked about eleven sleep habits. They were also divided into four groups according to age and job type: 23-29 (n = 226), 30 39 (n = 597), 40-49 (n = 225) and 50-59 (n = 113); managerial (n = 209), professional (n = 336), technical (n = 475) and clerical (n = 134). In this study, the sleep problems were defined as who had at least one of the following four poor sleep habits: 1) taking more than 30 minutes to fall asleep, 2) awakening during sleep (ADS) almost every day, 3) early morning awakening (EMA) almost every day, and 4) excessive daytime sleepiness (EDS) at work almost every day. Analyses showed that sleep problems were present in 26.0% of workers; the problems were most serious among workers aged 50 years and more (36.0%) which may be due to a significant increase in the prevalence rate of EMA. The results revealed that sleep problems are common in male white-collar daytime workers. The findings also suggest a need for increased attention to sleep problems in older workers and younger ones with EDS at work. PMID- 10680313 TI - Laboratory measurement of hazardous fumes and gases at a point corresponding to breathing zone of welder during a CO2 arc welding. AB - Concentrations of fumes, ozone (O3), carbon monoxide (CO), nitric oxide (NO), manganese (Mn) and total and hexavalent chromium (Cr) as well as size distribution of fumes were measured at a point corresponding to the welder's breathing zone during CO2-arc welding, using a welding robot and three kinds of wires. Concentrations of fumes, O3, CO, Mn and total-Cr were found to exceed their corresponding occupational exposure limit (OEL) values, while the concentrations of NO and Cr(VI) were below those OEL levels. Airborne concentration of Mn exceeded its OEL value, and the Mn content was 8 times higher in welding fumes than in the wire. Using an additive equation of OEL and exposure concentration of each hazardous component, health risk in welders with combined exposure to welding fumes and gases was assessed as 18.6 to 46.0 times of OEL, which exceeded the unity. This finding suggests that effective protection of welders from the exposure can be attained by use of the supplied-air respirator or combined use of a dust respirator and a local exhaust system. PMID- 10680314 TI - Changes in cold-induced vasodilatation, pain and cold sensation in fingers caused by repeated finger cooling in a cool environment. AB - To examine how repeated cooling of fingers with a rest pause schedule at work affects cold-induced vasodilatation (CIVD), pain and cold sensation in fingers, six healthy men aged 21 to 23 years immersed their left index fingers six times in stirred water at 10 degrees C for 10 minutes. After each cold-water immersion of the fingers, 5-minute rest pause was taken to observe the recovery process of the indicators. This cold-water immersion/rest pause test was carried out in a range of three ambient temperature conditions: 30 degrees C (warm), 25 degrees C (thermoneutral), and 20 degrees C (cool) as experienced in daily life. At the ambient temperatures of 30 degrees C and 25 degrees C, marked CIVD response occurred and the CIVD reactivity did not significantly change upon repetition of cold-water immersion. The lowered finger skin temperature also tended to recover quickly to the pre-immersion level during each post-immersion rest period. At the ambient temperature of 20 degrees C, however, the CIVD response weakened continuously upon repetition of immersion and almost disappeared during the final immersion. The recovery of finger skin temperature during each post-immersion rest was gradually delayed upon repetition of immersion. At every ambient temperature, finger pain and cold sensation induced by each cold-water immersion significantly decreased upon repetition of immersion and completely disappeared during each post-immersion rest period. Oral temperature during the experiment showed no significant change at the ambient temperatures of 25 degrees C and 30 degrees C, but it decreased significantly at the ambient temperature of 20 degrees C. These results suggest that in a cool work environment where the body core temperature is liable to decrease, repeated finger cooling may weaken CIVD reactivity and delay the recovery of finger temperature during post-immersion rest periods. In such lower ambient temperature work conditions, subjective judgements such as the decrease in finger pain and cold sensation during repeated finger cooling and the absence of them during post-immersion rest may not be reliable indicators for monitoring the risk of progressive tissue cooling and frostbite formation. PMID- 10680315 TI - Individual metal responsive elements of the human metallothionein-IIA gene independently mediate responses to various heavy metal signals. AB - Metallothioneins (MTs) are small metal-binding proteins that have a role in the defense against heavy metals. Mammalian MT genes are transcriptionally activated by metals such as Cd and Zn through multiple copies of the metal responsive element (MRE) present in the 5'-flanking region. To examine whether each MRE in a single promoter has a distinct role, we characterized seven MREs located upstream of the human MT-IIA gene. By transient transfection experiments using MRE-driven reporter gene constructs, individual MREs were assayed for the activity to mediate transcription in response to several heavy metal species. Four MREs including MREs a, b, e and g independently mediated reporter gene expression in response to Zn, Cd and Hg, while other MREs were not responsive to any of these metals. These results suggest that the multiplicity of MRE contributes to enhancing its activity, rather than providing functional diversity. PMID- 10680316 TI - Selective determination method for vanadium (V) and vanadium (IV) controlling the pH of media for a solid-liquid extraction column. AB - Solid-liquid extraction separation based on a speciation method was studied for selective determination of vanadium (IV) and vanadium (V). Both V (IV) and V (V) cations transform to oxo-acid anion along with pH changes in the solution. The pH values for the transition points are different from each other and the difference was utilized in the separation. In the first step, particulate samples are dissolved by strong acids such as 1 M H2SO4 or 85% H3PO4. The pH of the strong acidic sample solutions of V (IV) and V (IV) are adjusted to the range between 3 and 4. In this condition, V (IV) is in the form of cation but V (V) is anion. The pH adjusted solution is applied to an anion exchange column. The solution is expected to contain V (IV) only. The trapped V (V) anion is eluted as VO2+ cation by a pH 1 acid. The author and coworker have already developed an HPLC separation method utilizing this separation concept. However, the HPLC method has some limitations, mainly originated in physical and chemical weaknesses of the HPLC column. In the present study, a firm solid-liquid column is adopted to replace the feeble HPLC column as a separation device. And a simple and convenient pH adjustment technique for making the sample solution is investigated at the same time. With these improvements, the speciation method developed with strong acidic solutions could determine the amount of V (IV) and V (V) in various environmental and biological samples. PMID- 10680317 TI - Relationship between blood lead level and urinary ALA level in workers exposed to very low levels of lead. AB - The relationship between blood lead (PbB) level and urinary delta-aminolevulinic acid (ALAU) level was examined in a total of 3,636 lead-exposed workers in a periodic medical examination in 1992, in accordance with the Ordinance on Prevention of Lead Poisoning. The results were consistent with previously reported results in that ALAU level was found to increase with an increase in PbB level above 22.4 micrograms/dl (1.35 as a logarithmic value) and to rise markedly above 35.5 micrograms/dl (1.55). On the contrary, the geometric means of ALAU levels appeared to decrease with an increase in PbB levels within a range between a logarithmic value of 0.15 (1.4 micrograms/dl) and 1.25 (17.8 micrograms/dl). Because the earliest sign of the adverse health effects of lead is reported to occur at a PbB level of of 20 micrograms/dl, the relationship between PbB level and ALAU level was examined at PbB levels below 20 micrograms/dl. A regression formula was obtained, Y (log ALAU (mg/l)) = -0.0570X (log PbB (microgram/dl) + 0.4099. This result indicates that ALAU level decreases with a concomitant increase in PbB level lower than 20 micrograms/dl. PMID- 10680318 TI - Effects of coffee consumption against the development of liver dysfunction: a 4 year follow-up study of middle-aged Japanese male office workers. AB - The association of coffee consumption with the development of increased serum aspartate aminotransferase (AST) and/or alanine aminotransferase (ALT) activities over 4 years was studied in 1221 liver dysfunction-free (serum AST and ALT < or = 39 IU/l and no medical care for or no past history of liver disease) Japanese male office workers aged 35 to 56 years. From the analysis using the Kaplan-Meier method, the estimated incidence of serum AST and/or ALT > or = 40 IU/l, > or = 50 IU/l, and > or = 60 IU/l decreased with an increase in coffee consumption. From the Cox proportional hazards model, coffee drinking was independently inversely associated with the development of serum AST and/or ALT > or = 40 IU/l (p = 0.019 by test for tendency), > or = 50 IU/l (p = 0.002), and > or = 60 IU/l (p = 0.007), controlling for age, body mass index, alcohol intake, and cigarette smoking. These results suggest that coffee may be protectively against the liver dysfunction in middle-aged Japanese men. PMID- 10680319 TI - On "A retrospective cohort study of male workers exposed to PVA fibers". PMID- 10680320 TI - OSMA launches physicians' campaign for a healthier Oklahoma. PMID- 10680321 TI - A cardiac patient with renal failure: a clinicopathologic correlation conference from the University of Oklahoma College of Medicine. PMID- 10680322 TI - Immunization rate comparison for Oklahoma Medicaid managed care population by age two: 1995-1998. AB - Processes of care are used as one measure for the quality of care rendered by providers. One example is the immunization of children by the age of two. The Oklahoma Health Care Authority, with the Oklahoma Foundation for Medical Quality, has been tracking childhood immunization rates from 1995 through 1998. The rate calculated included the medical record data and the Oklahoma Statewide Immunization Information System (OSIIS) data set. The standards were based on Advisory Committee on Immunization Practices recommendations and Quality Assurance Reform Initiative standards. The rate of provision of documented immunizations in the Medicaid managed care population under the age of two has improved markedly from 1995 to 1998. The trend analysis suggests there might be areas for continued improvement in the provision of immunization to individuals in Oklahoma's Medicaid managed care population. Further, the OSIIS data is critical for maintaining a uniform data set for immunization information. PMID- 10680323 TI - Severe invasive group A beta-hemolytic streptococcus infection complicating pharyngitis: a case report and discussion. AB - Group A beta-hemolytic streptococcus (GABHS) has long been recognized as a deadly pathogen with manifestations ranging from impetigo to necrotizing fasciitis. Bacteremia from streptococcal pharyngitis is a rare complication. We report a patient presenting with septic shock and diabetic ketoacidosis from streptococcal pharyngitis. The pathophysiology, classification, and treatment of invasive group A streptococcal infection is discussed. PMID- 10680324 TI - The Internet for dentists. PMID- 10680325 TI - Biopsychosocial solutions to TMD. PMID- 10680326 TI - Probationary adhesive dentistry. PMID- 10680327 TI - Glass ionomer restorations. PMID- 10680328 TI - Considerations re: Antibiotic prophylaxis for dental patients at risk. Canadian Dental Association. PMID- 10680329 TI - The oral effects of smokeless tobacco. AB - Smokeless tobacco use has increased rapidly in North America. This form of tobacco use has many oral effects including leukoplakia, oral cancer, loss of periodontal support (recession), and staining of teeth and composite restorations. Systemic effects such as nicotine dependence, transient hypertension and cardiovascular disease may also result from smokeless tobacco use. This paper aims to guide dental practitioners in identifying oral lesions that occur due to the use of smokeless tobacco and also offer guidelines on how to counsel patients who express a desire to stop using smokeless tobacco products. PMID- 10680330 TI - [Familial adenomatous polyposis or Gardner syndrome--review of the literature and presentation of 2 clinical cases]. AB - Gardner syndrome is a type of hereditary gastrointestinal polyposis. Dental professionals should be aware that this syndrome can present as multiple impacted teeth and sometimes as large osteomas in the head and neck area. Following a brief review of literature, we present two cases of Gardner syndrome. One of these cases was diagnosed after a dental examination. The high incidence of malignant transformation of polyps into colorectal cancer indicates the importance of early diagnosis and follow-up. PMID- 10680331 TI - When professional burnout syndrome leads to dysthymia. PMID- 10680332 TI - Effects of composite thickness on the shear bond strength to dentin. AB - The manufacturers of some condensable posterior composites claim that their products can be placed in bulk and light-cured in 5-mm-thick increments. This study compared the shear bond strengths of three composite resins when bonded to dentin in 2- and 5-mm-thick increments. Overall the bond strengths were adversely affected by the composite thickness (p < 0.0001). The shear bond strength of each composite tested was much lower when polymerized in a 5-mm increment than in a 2 mm increment of composite (p < or = 0.0005). The two condensable composites tested had a lower bond strength than the conventional composite when polymerized in a 5-mm bulk increment (p < or = 0.01). PMID- 10680333 TI - Risk of recurrence of fetal chromosomal aberrations: analysis of trisomy 21, trisomy 18, trisomy 13, and 45,X in 1,076 Japanese mothers. AB - OBJECTIVE: To evaluate the risk of recurrence of fetal chromosomal aberrations in women who had offspring with numeric chromosomal abnormalities. SUBJECTS AND METHODS: This collaborative study consisted of 1,076 Japanese women with a history of offspring with trisomy-21, -18, -13, or 45,X. Second-trimester amniocenteses were performed, resulting in 1,248 fetal karyotypes that were analyzed with reference to prior offspring karyotypes and maternal age. RESULTS: Of the 842 women with trisomy-21 offspring, 10 conceived another such fetus. In 2 women with 3 or more such offspring, parental mosaicism of trisomy-21 was suspected. The incidence of recurrence of trisomy-21 increased with age, and significantly exceeded the incidence of trisomy-21 fetuses in the general population. None of the 170 women with trisomy-18 offspring, and none of the 46 women with trisomy-13 offspring, had another such fetus. Of the 18 women with 45,X offspring, 1 with mos 45,X/46,XX had another such fetus. CONCLUSIONS: The risk of recurrence of trisomy-21 is affected by maternal age and parental germline mosaicism. The risk of recurrence of trisomy-18 or -13 appears to be much lower than that of trisomy-21. Women who give birth to more than 1 offspring with 45,X should be examined for mos 45,X/46,XX. PMID- 10680334 TI - Microsatellite instability and hMSH2 gene mutation in a triple cancer (colon cancer, endometrial cancer, ovarian cancer) patient in hereditary non-polyposis colorectal cancer (HNPCC) kindred. AB - A patient who had triple cancer (colon cancer, endometrial cancer, and ovarian cancer) in HNPCC kindred is reported. Her family history revealed the occurrence of colon cancer in her paternal aunt and in two cousins, fulfilling the minimum HNPCC criteria. Microsatellite instability analysis revealed replication error (RER)+ in all cancer lesions at 2 microsatellite loci (D1S191, BAT 40). SSCP analysis suggested germline mutation in exon 2 of the hMSH2 gene. This case showed the importance of complete family-history investigations to identify HNPCC patients. In the near future, definitive diagnosis of HNPCC will be possible on the basis of DNA studies. PMID- 10680335 TI - Significance of multi-drug-resistant proteins in predicting chemotherapy response and prognosis in epithelial ovarian cancer. AB - OBJECTIVES: To clarify the expression of multi-drug-resistant (MDR) markers, GST pi, c-Jun, P-glycoprotein (Pgp), and MDR-associated protein (MRP) in epithelial ovarian cancer, and to determine whether their expression is predictive of chemotherapy response and patient prognosis. METHODS: Specimens of 58 epithelial ovarian cancer cases obtained at initial surgery were studied immunohistochemically using antibodies. RESULTS: Overall positive rates in the 58 specimens were 58.6% for GST-pi, 44.8% for c-Jun, 27.6% for Pgp, and 22.4% for MRP. The 5-year disease-free survival rate was 26.0% for patients with MRP positive tumors and 75.2% for those with MRP-negative tumors. The prognosis for those with MRP-positive tumors was significantly poorer (p < 0.05). Patients with GST-pi-positive tumors had a significantly worse prognosis than those with GST-pi negative tumors (51.9% vs 79.2%, p < 0.05). Multivariate analysis showed that residual tumors 2 cm or larger and MRP expression were independent prognostic factors for chemotherapy resistance. The relative risk of chemotherapy resistance in a patient with a residual tumor 2 cm or larger, positive MRP, and positive GST pi was 10.6 times greater than the risk in a patient without these factors. CONCLUSION: MRP and GST-pi expression might be potential predictors of the response to standard chemotherapy in epithelial ovarian cancer. Their expression also might contribute to individualizing clinical trials of postoperative chemotherapy. PMID- 10680336 TI - Clinicopathological study of recurrent uterine cervical squamous-cell carcinoma. AB - OBJECTIVE: To improve prognoses of patients with recurrent uterine cervical squamous-cell carcinoma. PATIENTS AND METHODS: We clinicopathologically analyzed 464 patients with uterine cervical squamous-cell carcinoma (126 positive, 338 negative pelvic lymph-node metastasis) who were treated at the Saitama Cancer Center from January 1, 1976 to December 31, 1991. RESULTS: The recurrence rates of negative pelvic lymph-node metastasis patients were 14. 2% (39/274) in pT1b and 32.8% (21/64) in pT2b. But for positive lymph-node metastasis patients the rates were 39.0% (23/59) in pT1b and 58.2% (39/67) in pT2b. The interval to recurrence was shorter in positive pelvic lymph-node patients than in negative patients. The 5-year survival rates after relapse of negative lymph-node patients with intrapelvic, extrapelvic, and both-sites recurrence were 53, 12, and 40%, respectively. But among distant recurrent sites, lung metastasis in negative lymph-node patients and lymphatic tract metastasis brought relatively fair prognoses. CONCLUSIONS: Regular long-term checks are necessary and active retreatments are recommended for patients with local recurrences, lung metastasis, or lymphatic vessel metastatic lesions. PMID- 10680337 TI - Recent trends in histological pattern of cervical carcinoma among three ethnic groups in Malaysia. AB - OBJECTIVE: To study the trend of different histological types of cervical carcinoma among the 3 major ethnic groups in Malaysia. METHODS: All invasive cervical carcinoma histologically diagnosed for the first time in 1991-1992 and 1996-1997 at the University Hospital, Kuala Lumpur (UHKL) were reviewed for the following parameters; age, ethnic group, histological category. RESULTS: One hundred and twenty-one and 145 cases were diagnosed in 1991-1992 and 1996-1997, respectively. During both periods, squamous was followed by adeno and adenosquamous carcinoma in frequency. Patients' mean ages ranged within the 4th decade for all 3 major histological types. Ethnically, an overall predilection for the Chinese was observed. While squamous carcinomas had declined among the Chinese and Malays, adenocarcinomas were noted to increase. The converse was observed among the Indians. CONCLUSIONS: Ethnically, cervical carcinoma showed a predilection for Malaysians of Chinese descent. A decreasing incidence of squamous with a worrying increasing trend of adenocarcinoma was observed, like in other populations studied. PMID- 10680338 TI - Brain natriuretic peptide (BNP) and cyclic guanosine monophosphate (cGMP) levels in normal pregnancy and preeclampsia. AB - OBJECTIVE: Our purpose was to evaluate plasma levels of brain natriuretic peptide (pBNP) and cyclic guanosine monophosphate (pcGMP) in preeclamptic patients and controls. STUDY DESIGN: Blood samples were obtained from 35 patients with preeclampsia and from the same women during the subsequent puerperal period. The control group consisted of normotensive pregnant women, matched with the patients for age, gestational age, and parity. The concentrations of pBNP and pcGMP were determined by the RIA method. Statistical analysis was performed using the Mann Whitney's U test. RESULTS: The pBNP level in the preeclampsia group was significantly increased, to 7-fold that of the control group. The pcGMP level was 50% higher in the preeclampsia group than in the control group, but this was not significant. Both the pBNP level and the pcGMP level in the puerperal period did not significantly differ between the patients and the controls. CONCLUSION: The pBNP concentrations increased in the preeclamptic women, and then these compensations were normalized in the puerperal period. PMID- 10680339 TI - Androgen-producing ovarian tumors: a clinicopathological study of 3 cases. AB - The clinical course and pathological findings of 3 rare cases of androgen producing ovarian tumors are presented. The ages of the 3 patients (Cases 1, 2, and 3, respectively) were 43, 34, and 57 years, respectively. Their preoperative serum testosterone levels were 506, 491, and 231 ng/dl, respectively. The pathological diagnoses of Cases 1, 2, and 3 were a Sertoli-stromal cell tumor of intermediate differentiation, a stromal tumor containing Leydig cells, and a stromal tumor with minor sex cord elements, respectively. Patient 1 experienced a recurrence, of a lesion at the vaginal stump 1 year and 2 months after the initial surgery. The clinical courses of Cases 2 and 3 have been non contributory. PMID- 10680340 TI - Causes of stillbirth: an analysis of 77 cases. AB - OBJECTIVE: To devise preventive measures for stillbirths, which account for more than 70% of perinatal deaths in Japan. METHODS: We retrospectively reviewed the medical records of 77 women with singleton pregnancies who gave birth to stillborn infants at > or = 30 weeks between 1979 and 1996 at our hospital. RESULTS: Major malformations were present in 21 (27%) of 77 infants, including 11 infants with anencephaly. Two infants (2.6%) were severely hydropic. Preeclampsia preceded the stillbirth and might have been an indirect cause of stillbirth in 21 (39%) of 54 women whose infants had normal formations. The cause of stillbirth in 33 non-preeclamptic women was unclear in 15 (28%), abruptio placentae in 9, fetal growth retardation in 3, the HELLP syndrome in 3, chorioamnionitis in 2, and cord accident in 1. Abruptio placentae also occurred in 9 of 21 preeclamptic patients. Thus, abruptio placentae was responsible for 18 (33%) of 54 stillborn infants with a grossly normal appearance. An autopsy was performed on only 13 (24%) of 54 infants with grossly normal appearance and did not provide new information relating to deaths. CONCLUSIONS: The causes of stillbirth were many and varied, with a large proportion having no obvious cause, although autopsies were underused. Increased monitoring for women with preeclampsia and early diagnosis and prompt delivery for women with abruptio placentae might be helpful in reducing the number of stillbirths. PMID- 10680341 TI - Emergency obstetric hysterectomies for postpartum haemorrhage. AB - OBJECTIVE: To review emergency obstetric hysterectomy in our unit, including the indications for and morbidity associated with the procedure. DESIGN: A retrospective cases analysis. SUBJECTS AND METHODS: Records of obstetric patients who had undergone emergency hysterectomies in between 15 October 1993 and 31 December 1997 were reviewed retrospectively. RESULTS: There were 15,474 deliveries and 7 emergency obstetric hysterectomies. All cases had total abdominal hysterectomy. The indications for hysterectomy were uterine atony and placental disorders. There were one case of urinary bladder injury and 2 cases of disseminated intravascular coagulopathy. There was no maternal mortality. CONCLUSION: Emergency obstetric hysterectomy remains a potentially life-saving procedure in unavoidable catastrophe. The 7 patients with life threatening postpartum haemorrhage underwent hysterectomy after failure of conservative measures. The morbidity is low and there was no mortality in this series. PMID- 10680342 TI - The rapid biophysical profile for assessment of fetal well-being. AB - OBJECTIVE: To determine the efficacy of the rapid biophysical profile (BPP), the combination of amniotic fluid index (AFI) and sound-provoked fetal movement (SPFM) detected by ultrasound, in predicting intrapartum fetal distress in high risk pregnancies, compared with the nonstress test (NST). STUDY DESIGN: The prospective study of diagnostic tests was conducted on a total of 1,069 high-risk singleton pregnancies, undergoing antepartum assessment of both the standard NST and the new rapid BPP, including AFI and SPFM detected by ultrasound. Intrapartum continuous fetal heart rate (FHR) monitoring was performed in all of them. The diagnostic indices of the NST and the rapid BPP was calculated in term of predicting intrapartum fetal distress. RESULTS: The rapid BPP was a reliable predictor of intrapartum fetal distress with higher sensitivity and specificity. Its accuracy was better than that of the NST. The incidence of fetal compromise among positive, equivocal, and negative tests of the rapid BPP are 78.57, 15.82 and 0.9%, respectively, whereas the incidence among nonreactive and reactive NST are 31.63 and 2.52%, respectively. CONCLUSIONS: The rapid BPP is an effective predictor of intrapartum fetal distress in high-risk pregnancies. It may suffice as an inexpensive and less time-consuming method of evaluating antepartum fetal well-being. PMID- 10680343 TI - Delayed-type hypersensitivity reaction to human menopausal gonadotrophin. PMID- 10680344 TI - Comparing lifetime emissions of natural gas and conventional fuel vehicles: an application of the generalized ANCOVA model. AB - New regulations and incentives are encouraging the use of clean, alternative fuel vehicles (AFVs) in urban areas. These vehicles are seen as one option for reducing air pollution from mobile sources. However, because of the limited number of AFVs on the road, little is known about actual lifetime emissions characteristics of in-use AFVs. This study describes the use of a generalized analysis of covariance model to evaluate and compare the emissions from natural gas vehicles with emissions from reformulated gasoline vehicles. The model describes fleet-wide emissions deterioration, while also accounting for individual vehicle variability within the fleet. This ability to measure individual vehicle variability can then be used to provide realistic bounds for the emissions deterioration in individual vehicles and the fleet as a whole. In order to illustrate the use of the model, the carbon monoxide, oxides of nitrogen (NOx), non-methane hydrocarbon (NMHC), and carbon dioxide emissions characteristics of a fleet of dedicated natural gas Dodge Ram vans and a fleet of dedicated reformulated gasoline Dodge Ram vans operating in the U.S. government fleet are explored. The analysis demonstrates the utility of the statistical method and suggests a potential for natural gas Dodge Ram vans to be generally cleaner than their conventional fuel counterparts. However, in the case of NOx and NHMCs, the analysis also suggests that these emissions benefits might be reduced over the vehicle lifetime due to higher emissions deterioration rates for natural gas vehicles. As this paper is aimed at illustrating the analysis of the covariance model, the results reported herein should be considered within the context of a more comprehensive study of these data before general conclusions are possible. Generalization of these findings to other vehicle models and alternative fuel technologies is not justified without further study. PMID- 10680345 TI - An alternative to additional SO3 injection for fly ash conditioning. AB - Small concentrations, approximately 2-10 parts per million (ppm), of injected sulfur trioxide (SO3) have improved particulate collection efficiencies of electrostatic precipitators burning lower-sulfur coal. However, the addition of extra SO3 not only incurs costs but also presents negative environmental effects. This work explored a method that could be applied to existing coal-fired power plants to convert the sulfur dioxide (SO2) already present in the flue gas to sufficient levels of SO3 for fly ash conditioning as an alternative to adding SO3 by burning elemental sulfur. During this research, a pre-mixed natural gas flame was used to promote the conversion of SO2 to SO3 in a drop-tube furnace with average non-flame, free stream gas temperatures of 450 and 1000 K. SO3 concentrations measured by wet chemistry and confirmed using elemental balances of other sulfur species measured by gas chromatography revealed that as much as 7% of SO2 was homogeneously transformed to SO3. The results also showed that at low temperatures, the rate at which SO3 is converted back to SO2 decreased, thus extending the time period during which SO3 concentrations would be sufficient for ash conditioning. An additional benefit of this technique is speculated to result from increased flue gas humidity. PMID- 10680346 TI - Distance-weighted traffic density in proximity to a home is a risk factor for leukemia and other childhood cancers. AB - Occupational exposure to elevated concentrations of benzene is a known cause of leukemia in adults. Concentrations of benzene from motor vehicle exhaust could be elevated along highly trafficked streets. Several studies have reported significant associations between proximity to highly trafficked streets and the occurrence of childhood cancers and childhood leukemia. These associations may be due to chronic exposure to benzene or other carcinogenic components of vehicle exhaust from these nearby streets or to some other factor (e.g., noise, increased light exposure, or some unaccounted--for socioeconomic variable). We used data for homes studied in an earlier childhood cancer study conducted in Denver, CO, in the 1980s. No air pollution measurements were made in the original study. We identified the highest trafficked street near each study home and obtained the traffic density in 1979 and 1990. Traffic density was weighted for the distance from the street to the home using 3 different widths of Gaussian curves to approximate the decay of the emissions into the surrounding neighborhoods. The associations between the 750-ft-wide distance-weighted traffic density metrics and all childhood cancers and childhood leukemia are strongest in the highest traffic density category (> or = 20,000 vehicles per day [VPD]). The odds ratio is 5.90 (95% confidence interval [CI] 1.69-20.56) for all cancers and 8.28 (95% CI 2.09-32.80) for leukemia. The results are suggestive of an association between proximal high traffic streets with traffic counts > or = 20,000 VPD and childhood cancer, including leukemia. PMID- 10680347 TI - Determination of PM10 emission rates from street sweepers. AB - The use of street sweepers to clean paved roads, particularly after high-wind events, has been proposed as a PM10 control method. Using an artificial tunnel, the emission rates for several street sweepers were quantified under actual operating conditions. The tunnel was a tent enclosure, 6.1 x 4.3 x 73 m, open on both ends. PM10 concentrations were measured at the inlet and outlet while a sweeper removed sand deposited along the length. Measurements were made using a specialized low-volume filter sampler and an integrating nephelometer. The volume of air passing through the tunnel was measured by releasing an inert tracer, sulfur hexafluoride, at the inlet and measuring its concentration at the outlet. A large difference in emission rates between vacuum-type sweepers was observed, with rates varying from 5 to 100 mg m-1 swept. For the cleanest sweepers, the background rates (collected by sweeping clean pavement) were about half of the total PM10 emission rate. These background emission rates likely were from diesel exhaust; background rates for the single gasoline-powered sweeper were below detection. Particle light scattering data confirmed the filter collection results. The artificial tunnel approach would be useful in measuring total emissions from other mobile and stationary sources. PMID- 10680348 TI - Contribution of liquefied petroleum gas to air pollution in the metropolitan area of Mexico City. AB - An estimation of hydrocarbon emissions caused by the consumption of liquefied petroleum gas (LPG) in the Metropolitan Area of Mexico City (MAMC) is presented. On the basis of experimental measurements at all points of handling, during the distribution process, and during the consumption of LPG in industrial devices and domestic appliances, an estimated 76,414 tons/year are released to the air. The most important contribution is found during the domestic consumption of LPG (70%); this makes the control initiatives available to the consumer. By developing a control program of LPG losses, a 77% reduction in emission is expected in a 5-yr period. The calculated amounts of LPG emissions when correlated with the consumption of LPG, combined with information from air samples from the MAMC, do not point to LPG emissions as the most important factor contributing to tropospheric ozone in the air in Mexico City. PMID- 10680349 TI - Evaluation of total volatile organic compound emissions from adhesives based on chamber tests. AB - In 1997, Homeswest in western Australia and Murdoch University developed a project to construct low-allergen houses (LAHs) in a newly developed suburb. Before the construction of LAHs, all potential volatile organic compound (VOC) emission materials used in LAHs are required to be measured to ensure that they are low total VOC (TVOC) emission materials. This program was developed based on this purpose. In recent times, the number of complaints about indoor air pollution caused by VOCs has increased. A number of surveys of indoor VOCs have indicated that many indoor materials contribute to indoor air pollution. Although some studies have been conducted on the characteristics of VOC emissions from adhesives, most of them were focused on VOC emissions from floor adhesives. Few measurements of VOC emissions from adhesives used for wood, fabrics, and leather are available. Furthermore, most research on VOC emissions from adhesives has been done in countries with cool climates, where ventilation rates in the indoor environment are lower than those in Mediterranean climates, due to energy conservation. VOCs emitted from adhesives have not been sufficiently researched to prepare an emission inventory to predict indoor air quality and to determine both exposure levels for the Australian population and the most appropriate strategies to reduce exposure. An environmental test chamber with controlled temperature, relative humidity, and airflow rate was used to evaluate emissions of TVOCs from three adhesives used frequently in Australia. The quantity of TVOC emissions was measured by a gas chromatography/flame ionization detector. The primary VOCs emitted from each adhesive were detected by gas chromatography/mass spectrometry. The temporal change of TVOC concentrations emitted from each adhesive was tested. A double-exponential equation was then developed to evaluate the characteristics of TVOC emissions from these three adhesives. With this double-exponential model, the physical processes of TVOC emissions can be explained, and a variety of emission parameters can be calculated. These emission parameters could be used to estimate real indoor TVOC concentrations in Mediterranean climates. PMID- 10680350 TI - In-stack condensible particulate matter measurements and issues. AB - Particulate matter (PM) emitted from fossil fuel-fired units can be classified as either filterable or condensible PM. Condensible PM typically is not measured because federal and most state regulations do not require sources to do so. To determine the magnitude of condensible PM emissions relative to filterable PM emissions and to better understand condensible PM measurement issues, a review and analysis of actual U.S. Environmental Protection Agency (EPA) Method 202 (for in-stack condensible PM10) and EPA Method 201/201A (for in-stack filterable PM10) results were conducted. Methods 202 and 201/201A results for several coal-burning boilers showed that the condensible PM, on average, comprises approximately three fourths (76%) of the total PM10 stack emissions. Methods 202 and 201/201A results for oil- and natural gas-fired boilers showed that the condensible PM, on average, comprises 50% of the total PM10 stack emissions. Methods 202 and 201/201A results for oil-, natural gas-, and kerosene-fired combustion turbines showed that the condensible PM, on average, comprises 69% of the total PM10 stack emissions. Based on these limited measurements, condensible PM can make a significant contribution to total PM10 emissions for fossil fuel-fired units. A positive bias (indicating more condensible PM than is actually emitted) may exist in the measured data due to the conversion of dissolved sulfur dioxide to sulfate compounds in the sampling procedure. In addition, these Method 202 results confirm that condensible PM, on average, is composed mostly of inorganic matter, regardless of the type of fuel burned. PMID- 10680351 TI - Forecasts using neural network versus Box-Jenkins methodology for ambient air quality monitoring data. AB - This study explores ambient air quality forecasts using the conventional time series approach and a neural network. Sulfur dioxide and ozone monitoring data collected from two background stations and an industrial station are used. Various learning methods and varied numbers of hidden layer processing units of the neural network model are tested. Results obtained from the time-series and neural network models are discussed and compared on the basis of their performance for 1-step-ahead and 24-step-ahead forecasts. Although both models perform well for 1-step-ahead prediction, some neural network results reveal a slightly better forecast without manually adjusting model parameters, according to the results. For a 24-step-ahead forecast, most neural network results are as good as or superior to those of the time-series model. With the advantages of self-learning, self-adaptation, and parallel processing, the neural network approach is a promising technique for developing an automated short-term ambient air quality forecast system. PMID- 10680352 TI - Kinetics of catalytic oxidation of benzene, n-hexane, and emission gas from a refinery oil/water separator over a chromium oxide catalyst. AB - With the advances made in the past decade, catalytic incineration of volatile organic compounds (VOCs) has become the technology of choice in a wide range of pollution abatement strategies. In this study, a test was undertaken for the catalytic incineration, over a chromium oxide (Cr2O3) catalyst, of n-hexane, benzene, and an emission air/vapor mixture collected from an oil/water separator of a refinery. Reactions were carried out by controlling the feed stream to constant VOC concentrations and temperatures, in the ranges of 1300-14,700 mg/m3 and 240-400 degrees C, respectively. The destruction efficiency for each of the three VOCs as a function of influent gas temperature and empty bed gas residence time was obtained. Results indicate that n-hexane and the oil vapor with a composition of straight- and branch-chain aliphatic hydrocarbons exhibited similar catalytic incineration effects, while benzene required a higher incineration temperature or longer gas retention time to achieve comparable results. In the range of the VOC concentrations studied, at a given gas residence time, increasing the operating temperature of the catalyst bed increased the destruction efficiency. However, the much higher temperatures required for a destruction efficiency of over 99% may be not cost-effective and are not suggested. A first-order kinetics with respect to VOC concentration and an Arrhenius temperature dependence of the kinetic constant appeared to be an adequate representation for the catalytic oxidation of these volatile organics. Activation energy and kinetic constants were estimated for each of the VOCs. Low temperature destruction of the target volatile organics could be achieved by using the Cr2O3 catalyst. PMID- 10680353 TI - Methane emissions from domestic waste management facilities in Jordan- applicability of IPCC methodology. AB - In this paper, methane emissions from municipal wastewater treatment plants and municipal solid waste (MSW) landfills in Jordan for 1994 have been estimated using the methodology developed by the Intergovernmental Panel on Climate Change (IPCC). For this purpose, the 14 domestic wastewater treatment plants in the country were surveyed. Generation rates and characterization of MSW components as well as dumping and landfilling practices were surveyed in order to estimate 1994 CH4 emissions from these sites. Locally available waste statistics were used in cases where those of the IPCC guidelines were not representative of Jordan's statistics. Methane emissions from domestic wastewater in Jordan were estimated at 4.66 gigagrams (Gg). Total 1994 CH4 emissions from MSW management facilities in Jordan are estimated at 371.76 Gg--351.12 Gg (94.45%) from sanitary landfills, 19.83 Gg (5.33%) from MSW open dumps, and 0.81 Gg (0.22%) from raw sewage-water dumping ponds. Uncertainties associated with these estimations are presented. PMID- 10680354 TI - An intelligent emissions controller for fuel lean gas reburn in coal-fired power plants. AB - The application of artificial intelligence techniques for performance optimization of the fuel lean gas reburn (FLGR) system is investigated. A multilayer, feedforward artificial neural network is applied to model static nonlinear relationships between the distribution of injected natural gas into the upper region of the furnace of a coal-fired boiler and the corresponding oxides of nitrogen (NOx) emissions exiting the furnace. Based on this model, optimal distributions of injected gas are determined such that the largest NOx reduction is achieved for each value of total injected gas. This optimization is accomplished through the development of a new optimization method based on neural networks. This new optimal control algorithm, which can be used as an alternative generic tool for solving multidimensional nonlinear constrained optimization problems, is described and its results are successfully validated against an off the-shelf tool for solving mathematical programming problems. Encouraging results obtained using plant data from one of Commonwealth Edison's coal-fired electric power plants demonstrate the feasibility of the overall approach. Preliminary results show that the use of this intelligent controller will also enable the determination of the most cost-effective operating conditions of the FLGR system by considering, along with the optimal distribution of the injected gas, the cost differential between natural gas and coal and the open-market price of NOx emission credits. Further study, however, is necessary, including the construction of a more comprehensive database, needed to develop high-fidelity process models and to add carbon monoxide (CO) emissions to the model of the gas reburn system. PMID- 10680355 TI - A new screening measure to identify potential carbon monoxide hotspots. AB - To demonstrate conformity of transportation projects to National Ambient Air Quality Standards in accordance with State Implementation Plans, the U.S. Environmental Protection Agency (EPA) uses intersection level of service (LOS) as one of its major criteria for screening for potential carbon monoxide (CO) hotspots. Although intersection LOS is a measure of traffic volume, signal timing, and related congestion and delay, the assigned level reflects only the computed averaged stopped delay (ASD) per vehicle at the intersection. Thus, intersections can often operate at the same LOS but produce vastly different levels of predicted CO concentrations. For example, a two-lane approach operating at LOS D will produce very different levels of CO than a five-lane approach also operating at LOS D. This study explores the effectiveness of the LOS D criterion as a screen for identifying potential CO hotspots. The study results indicate that LOS is a poor predictor of potential CO hotspots when compared to results generated with the EPA-recommended micro-scale model CAL3QHCr. To more consistently screen out those intersections that will not be identified as CO hotspots using the micro-scale models, a new criterion, equivalent red-time vehicles (ERTV), is introduced. The modeling results using ERTV suggest that it is a more robust measure for identifying potential CO hotspots, and conversely, screening out those intersections that are not likely to be identified as hotspots using micro-scale simulation results. PMID- 10680356 TI - Meteorological factors of ozone predictability at Houston, Texas. AB - Several ozone modeling approaches were investigated to determine if uncertainties in the meteorological data would be sufficiently large to limit the application of physically realistic ozone (O3) forecast models. Three diagnostic schemes were evaluated for the period of May through September 1997 for Houston, TX. Correlations between measured daily maximum and model calculated O3 air concentrations were found to be 0.70 using a linear regression model, 0.65 using a non-advective box model, and 0.49 using a three-dimensional (3-D) transport and dispersion model. Although the regression model had the highest correlation, it showed substantial underestimates of the highest concentrations. The box model results were the most similar to the regression model and did not show as much underestimation. The more complex 3-D modeling approach yielded the worst results, likely resulting from O3 maxima that were driven by local factors rather than by the transport of pollutants from outside of the Houston domain. The highest O3 concentrations at Houston were associated with light winds and meandering trajectories. A comparison of the gridded meteorological data used by the 3-D model to the observations showed that the wind direction and speed values at Houston differed most on those days on which the O3 underestimations were the greatest. These periods also tended to correspond with poor precipitation and temperature estimates. It is concluded that better results are not just obtained through additional modeling complexity, but there needs to be a comparable increase in the accuracy of the meteorological data. PMID- 10680357 TI - Pyrolysis kinetics and residue characteristics of petrochemical industrial sludge. AB - This study investigated the pyrolysis characteristics of sludge from wastewater treatment plants in the petrochemical industry and focused on the pyrolysis kinetics, elemental composition of residue, and volatile organic compounds (VOCs) of exhaust gas. As pyrolysis temperature increased to 773 K, the increasing rate of crude oil production tended to a stable condition. The result indicated that the optimal temperature of crude oil and water mixed production was 773 K. When pyrolysis temperature increased from 673 to 973 K, carbon, oxygen, nitrogen, and hydrogen concentrations of residue decreased and the sulfur concentration of residue increased. The concentrations of benzene, toluene,ethylbenzene, and styrene increased by the increasing pyrolysis temperature. We found that the reaction order of sludge pyrolysis was 2.5 and the activation energy of the reaction was 11.06 kJ/mol. We believe that our pyrolysis system is transitional between devolatilization and combustion. PMID- 10680358 TI - Real-world vehicle emissions: a summary of the Ninth Coordinating Research Council On-Road Vehicle Emissions Workshop. AB - In April 1999, the Coordinating Research Council sponsored a workshop focusing on our understanding of real-world emissions from motor vehicles. This summary presents the latest information on in-use light- and heavy-duty vehicle tailpipe and evaporative emissions, the effects of fuels on emissions, field programs designed to understand the contribution of mobile sources to emission inventories, efforts to evaluate and improve mobile source emission models, progress of vehicle inspection/maintenance programs, and topics for future research. While significant progress has been made in understanding in-use vehicle emissions, further improvements are necessary. Moreover, the impact of current and future changes in emission control technologies and control programs will have to be monitored for effectiveness and incorporated into the emission factor models. PMID- 10680359 TI - Modeling relationships between indoor and outdoor air quality. AB - Information about the ratio between indoor and outdoor concentrations (IO ratios) of air pollutants is a crucial component in human exposure assessment. The present study examines the relationship between indoor and outdoor concentrations as influenced by the combined effect of time patterns in outdoor concentrations, ventilation rate, and indoor emissions. Two different mathematical approaches are used to evaluate IO ratios. The first approach involves a dynamic mass balance model that calculates distributions of transient IO ratios. The second approach assumes a linear relationship between indoor and outdoor concentrations. We use ozone and benzene as examples in various modeling exercises. The modeled IO ratio distributions are compared with the results obtained from linear fits through plots of indoor versus outdoor concentrations. PMID- 10680360 TI - An evaluation of the LPG vehicles program in the metropolitan area of Mexico City. AB - The environmental agency in the metropolitan area of Mexico City has launched a program to introduce more energy-efficient modes of transport, one of which is the use of alternative and less polluting fuels. With the perspective in mind, a liquefied petroleum gas (LPG) fleet of vehicles is exempt of the mandatory "one day without a car" program if the emission of pollutants is below the standard authorized for that specific purpose. Today, about 28,000 light-duty vehicles and heavy-duty trucks circulate in the area, most of them as aftermarket converted vehicles. In this work, we evaluated regulated exhaust emission and other parameters on 134 representative vehicles of that fleet. From the data obtained, an estimate of emission factors and their contribution to the global emission in the metropolitan area is provided. It is concluded that more than 95% of the in use vehicles using LPG presented regulated emissions which exceeded in one or more the environmental regulations values required for certification. The poor maintenance of the vehicles and the type of conversion kit installed could be the culprits of the results obtained. PMID- 10680361 TI - A new dose model for assessment of health risk due to contaminants in air. AB - The problem of making quantitative assessments of the risks associated with human exposure to toxic contaminants in the environment is a pressing one. This study demonstrates the capability of a new computational technique involving the use of fuzzy logic and neural networks to produce realistic risk assessments. The systematic analysis of human exposure involves the use of measurements and models, the results of which are sometimes used in regulatory decisions or in the drafting of legislation. Because of limited scientific understanding, however, interpretation of models often involves substantial uncertainty. Extensive measurement programs can be very expensive. The high complexity and inherent heterogeneity of exposure analysis is still a major challenge. The approach to this challenge tested here is to use a new model incorporating sophisticated artificial intelligence algorithms. Exposure assessment often requires that a number of factors be evaluated, including exposure concentrations, intake rates, exposure times, and frequencies. These factors are incorporated into a system that can "learn" the relevant relationships based on a known data set. The results can then be applied to new data sets and thus be applied widely without the need for extensive measurements. In this analysis, an example is developed for human health risk through inhalation exposure to benzene from vehicular emissions in the cities of Auckland and Christchurch, New Zealand. Risk factors considered were inhaled contaminant concentration, age, body weight, and activity patterns of humans. Three major variables affecting the inhaled contaminant concentration were emissions (mainly from motor vehicles), meteorology (wind speed, temperature, and atmospheric stability), and site factors (hilly, flat, etc.). The results are preliminary and used principally to demonstrate the technique, but they are very encouraging. PMID- 10680362 TI - Modeling and direct sensitivity analysis of biogenic emissions impacts on regional ozone formation in the Mexico-U.S. border area. AB - A spatially and temporally resolved biogenic hydrocarbon and nitrogen oxides (NOx) emissions inventory has been developed for a region along the Mexico-U.S. border area. Average daily biogenic non-methane organic gases (NMOG) emissions for the 1700 x 1000 km2 domain were estimated at 23,800 metric tons/day (62% from Mexico and 38% from the United States), and biogenic NOx was estimated at 1230 metric tons/day (54% from Mexico and 46% from the United States) for the July 18 20, 1993, ozone episode. The biogenic NMOG represented 74% of the total NMOG emissions, and biogenic NOx was 14% of the total NOx. The CIT photochemical airshed model was used to assess how biogenic emissions impact air quality. Predicted ground-level ozone increased by 5-10 ppb in most rural areas, 10-20 ppb near urban centers, and 20-30 ppb immediately downwind of the urban centers compared to simulations in which only anthropogenic emissions were used. A sensitivity analysis of predicted ozone concentration to emissions was performed using the decoupled direct method for three dimensional air quality models (DDM 3D). The highest positive sensitivity of ground-level ozone concentration to biogenic volatile organic compound (VOC) emissions (i.e., increasing biogenic VOC emissions results in increasing ozone concentrations) was predicted to be in locations with high NOx levels, (i.e., the urban areas). One urban center- Houston--was predicted to have a slight negative sensitivity to biogenic NO emissions (i.e., increasing biogenic NO emissions results in decreasing local ozone concentrations). The highest sensitivities of ozone concentrations to on road mobile source VOC emissions, all positive, were mainly in the urban areas. The highest sensitivities of ozone concentrations to on-road mobile source NOx emissions were predicted in both urban (either positive or negative sensitivities) and rural (positive sensitivities) locations. PMID- 10680363 TI - Particulate matter modeling in the Los Angeles basin using SAQM-AERO. AB - The SARMAP air quality model, enhanced with aerosol modeling capability, and its associated components were developed to understand causes of ozone (O3) and particulate matter exceedances in the San Joaquin Valley of California. In order for this modeling system to gain increasing acceptance and use in guiding air quality management, it is important to assess how transportable this modeling system is across geographic domains. We describe the first application of the modeling system outside the "home" domain for which it was developed and evaluated. We have chosen the August 27-28, 1987, intensive monitoring period of the Southern California Air Quality Study to evaluate the performance of the modeling system and to assess its sensitivity to emission control options. The predicted surface concentrations of O3 and other gas-phase species were spatially and temporally correlated with measured data. The fractional normalized absolute error was 0.32 to 0.36 for O3, and somewhat larger for other species. The fractional normalized bias for O3 on August 27 and 28, 1987, was 0.02 to 0.04. The simulated PM2.5 mass and constituent species concentrations reproduced the magnitude and variability of the observed daytime concentrations at most locations; however, nighttime PM2.5 concentrations were overpredicted by the model. The model's response to emission control options was consistent with other models of the same genre. PMID- 10680364 TI - Trends in fine particle concentration and chemical composition in southern California. AB - Airborne fine particle mass concentrations in Southern California have declined in recent years. Trends in sulfate and elemental carbon (EC) particle concentrations over the period 1982-1993 are consistent with this overall improvement in air quality and help to confirm some of the reasons for the changes that are seen. Fine particle sulfate concentrations have declined as a strict sulfur oxides (SOx) emission control program adopted in 1978 was implemented over time. Fine particle elemental (black) carbon concentrations have declined over a period when newer diesel engines and improved diesel fuels have been introduced into the vehicle fleet. Organic aerosol concentrations have not declined as rapidly as the EC particle concentrations, despite the fact that catalyst-equipped cars having lower particle emission rates were introduced into the vehicle fleet alongside the diesel engine improvements mentioned above. This situation is consistent with the growth in population and vehicle miles traveled in the air basin over time. Fine particle ammonium nitrate in the Los Angeles area atmosphere contributes more than half of the fine aerosol mass concentration on the highest concentration days of the year, emphasizing both the need for accurate aerosol nitrate measurements and the likely importance of deliberate control of aerosol nitrate as a part of any serious further fine particle control program for the Los Angeles area. PMID- 10680365 TI - Climatology of diurnal trends and vertical distribution of ozone in the atmospheric boundary layer in urban North Carolina. AB - Vertical measurements of ozone were made on a 610-m-tall tower located about 15 km southeast of Raleigh, NC, as part of an effort by the state of North Carolina to develop a state implementation plan (SIP) for ozone control in the Raleigh Metropolitan Statistical Area. During summer 1993, 1994, and 1995, ozone was monitored at ground level, 250 m, and 433 m. Boundary layer wind, temperature, and other meteorological variable profiles were determined from balloon soundings. During summer 1996 and 1997, ozone was monitored at ground level, 76 m, 128 m, and 433 m. This paper presents the analysis and discussion of the five year data. The evolutions of the convective boundary layer during daytime and the stable nocturnal boundary layer (NBL) were found to have marked impacts on ozone concentrations. A strong diurnal pattern, with an afternoon maximum and an early morning minimum, was dominant at ground level, but it was much weaker at elevated levels and insignificant above the NBL at night. Ozone deposition velocities at night during the measurement periods were estimated to range from 0.09 to 0.64 cm/sec. We found evidence of regional transport of ozone and/or its precursors from northwest and north of the site, which may play a role in high ozone events in the Raleigh-Durham area. Ozone concentrations between the various elevated levels were well correlated, while correlations between the ground and upper levels were much weaker. However, a strong correlation was found between the nighttime and early morning ozone concentrations (CR) in the residual layer above the NBL and the maximum ground level concentration (Co max) the following afternoon. Based on this correlation, the latter may be predicted by an observational model Co max = 27.76e 0.016 CR. PMID- 10680366 TI - Assessing the impact of differential measurement error on estimates of fine particle mortality. AB - In air pollution epidemiology, error in measurements of correlated pollutants has been advanced as a reason to distrust regressions that find statistically significant weak associations. Much of the related debate in the literature and elsewhere has been qualitative. To promote quantitative evaluation of such errors, this paper develops an air pollution time-series model based on correlations among unit-normal variables. Assuming there are no other sources of bias present, the model shows the expected amount of relative bias in the regression coefficients of a bivariate regression of coarse and fine particulate matter measurements on daily mortality. The model only requires information on instrumental error and spatial variability, along with the observed regression coefficients and information on the true fine-course correlation. Analytical results show that if one pollutant is truly more harmful than the other, then it must be measured more precisely than the other in order not to bias the ratio of the fine and course regression coefficients. Utilizing published data, a case study of the Harvard Six-Cities study illustrates use of the model and emphasizes the need for data on spatial variability across the study area. Current epidemiology time-series regressions can use this model to address the general concern of correlated pollutants with differing measurement errors. PMID- 10680367 TI - Factors influencing the variability of SO2 concentrations in Istanbul. AB - The correlation between sulfur dioxide (SO2) concentrations measured at the European and Asian sides of Istanbul and meteorological parameters is investigated using principal component analysis (PCA) and multiple regression analysis techniques. Several meteorological parameters are selected to represent the atmospheric conditions during two winter periods: 1993-1994 and 1994-1995. Six principal components are found to explain the majority of the observed meteorological variability. Surface pressure, 850-mb temperature, and surface zonal (east-west) and meridional (north-south) winds show high loadings on separate factors identified by PCA. We seek dominant meteorological parameters that control the SO2 levels at each monitoring station. Several multiple regression analysis models are fitted to the data from each monitoring station using six principal components and previous-day SO2 concentrations as independent variables. Results suggest that the most important parameters, highly correlated with SO2 concentrations in the Istanbul metropolitan area, are atmospheric pressure and surface zonal and meridional winds. These components have more influence on the determination of the air pollution levels at the Asian side than at the European side. PMID- 10680368 TI - Aerosols in a Mediterranean forest: sulfates, particle size distribution, and growth rates. AB - Particle size distribution measurements in 16 nonlinear intervals covering the 0.1-7.5 microns range and concurrent sulfate concentrations were continuously recorded in September 1996 over the period of two weeks in a Mediterranean forest. Sulfate size distribution was derived from a linear correlation fit between the concentrations and the number of particles recorded at each particle size interval. The results revealed two modal diameters for sulfates, with typical diameters at 0.3 and 0.675 micron. These results were associated with two different dominant chemical mechanisms governing sulfate formation. In order to describe the dominant chemical mechanism, the growth law approach was applied. Growth rates were determined using the parameter estimates of the fitted particle size distribution function. By matching these data with sulfate concentrations, the dominant chemical reactions were identified. The results have shown that sulfate formation is governed by both homogeneous and heterogeneous reactions and that the latter process was dominant. Condensation reactions prevailed in the early morning and late afternoon, and volume reactions at night, particularly in high-moisture conditions. From the observational data, the gas-to-particle conversion rate for sulfur dioxide (SO2) at nighttime was also derived, yielding a 2.18%/hr-1. PMID- 10680369 TI - Particulates generated from combustion of polymers (plastics). AB - This is an experimental study on the characterization of particulate (soot) emissions from burning polymers. Emissions of polystyrene (PS), polyethylene (PE), polypropylene (PP), polymethyl methacrylate (PMMA), and polyvinyl chloride (PVC) plastics were studied. Combustion took place in a laboratory-scale, electrically heated, drop-tube furnace at temperatures of 1300 and 1500 K, in air. The nominal bulk (global) equivalence ratio, phi, was varied in the range of 0.5-1.5, and the gas residence time in the nearly isothermal radiation zone of the furnace was approximately 1 sec. The particulate emissions were size classified at the exit of the furnace, using a multi-stage inertial particle impactor. Results showed that both the yields and the size distributions of the emitted soot were remarkably different for the five plastics burned. Soot yields increased with an increasing bulk equivalence ratio. Combustion of PS yielded the highest amounts of soot (most highly agglomerated), several times more than the rest of the polymers. More soot was emitted from PS at 1500 than at 1300 K. Substantial amounts of soot agglomerates were larger than 9 microns. At 1500 and 1300 K, 35 and 29% of the soot mass, respectively, was PM2 (2 microns or smaller). Emissions from PE and PP were remarkably similar to each other. These polymers produced very low emissions at phi < or = 0.5, but emissions increased drastically with phi, and most of the soot was very fine (70-97% of the mass was PM2, depending on phi). Emissions from the combustion of PMMA were comparatively low and were the least influenced by the bulk phi, and 79-95% of the emissions were PM2. Combustion of PVC yielded the lowest amounts of soot; moreover, only 13 34% of the mass was PM2. On a comparative basis, at 1500 K, the following ranges of particulate yields were PM2: 19-75 mg/g of PS, 8-36 mg/g of PE, 1.5-47 mg/g of PP, 11-20 mg/g of PMMA, and 2-8 mg/g of PVC, depending on phi. These comparative results demonstrate that PS produces the highest amounts of fine particulates, followed by PP, PE, and PMMA, and then PVC. Burning these materials with excess oxygen drastically reduces the particulate emissions of PE and PP, substantially reduces those of PS, and mildly reduces those of PMMA and PVC. PMID- 10680370 TI - Indoor/outdoor relationships for ambient PM2.5 and associated pollutants: epidemiological implications in Lindon, Utah. AB - Outdoor and indoor fine particulate species were measured at the Lindon Elementary School in Lindon, Utah, to determine which components of ambient fine particles have strong indoor and outdoor concentration correlations. PM2.5 mass concentrations were measured using tapered element oscillating microbalance (TEOM) monitors and by gravimetric analysis of Teflon filter samples. Gas-phase HNO3, sulfur dioxide, particulate nitrate, strong acid, and particulate sulfate were measured using annular denuder samplers. Soot was measured using quartz filters in filter packs. Total particulate number was measured with a condensation nucleus counter (CNC). Total particulate number and fine particulate sulfate and soot were correlated for ambient and indoor measurements. Indoor PM2.5 mass showed a low correlation with outdoor PM2.5 mass because of the influence of coarse material from student activities on indoor PM2.5. Fine particle acidity and the potentiation of biological oxidative mechanisms by iron were not correlated indoors and outdoors. PMID- 10680371 TI - Continuous emissions monitoring using spark-induced breakdown spectroscopy. AB - A new technology for monitoring airborne heavy metals on aerosols and particulates based on spark-induced breakdown spectroscopy (SIBS) was evaluated at a joint U.S. Environmental Protection Agency (EPA)/U.S. Department of Energy test at the rotary kiln incinerator simulator (RKIS) facility at EPA/Research Triangle Park, NC, in September 1997. The instrument was configured to measure lead and chromium in a simulated combustion flue gas in real time and in situ at target levels of 15 and 75 micrograms/dry standard cubic meters. Actual metal concentrations were measured during the tests using EPA Reference Method (RM) 29. The SIBS technology detected both lead and chromium at the low- and high-level concentrations. Additionally, the hardware performed without failure for more than 100 hr of operation and acquired data for 100% of the RM tests. The chromium data were well correlated with concentration increases resulting from duct operations and pressure fluctuations that are known to entrain dust. PMID- 10680372 TI - Near surface soil vapor clusters for monitoring emissions of volatile organic compounds from soils. AB - The overall objective of this research was to develop and test a method of determining emission rates of volatile organic compounds (VOCs) and other gases from soil surfaces. Soil vapor clusters (SVCs) were designed as a low dead volume, robust sampling system to obtain vertically resolved profiles of soil gas contaminant concentrations in the near surface zone. The concentration profiles, when combined with a mathematical model of porous media mass transport, were used to calculate the contaminant flux from the soil surface. Initial experiments were conducted using a mesoscale soil remediation system under a range of experimental conditions. Helium was used as a tracer and trichloroethene was used as a model VOC. Flux estimations using the SVCs were within 25% of independent surface flux estimates and were comparable to measurements made using a surface isolation flux chamber (SIFC). In addition, method detection limits for the SVC were an order of magnitude lower than detection limits with the SIFC. Field trials, conducted with the SVCs at a bioventing site, indicated that the SVC method could be easily used in the field to estimate fugitive VOC emission rates. Major advantages of the SVC method were its low detection limits, lack of required auxiliary equipment, and ability to obtain real-time estimates of fugitive VOC emission rates. PMID- 10680373 TI - Performance of the annular denuder system with different arrangements for HNO3 and HNO2 measurements in Taiwan. AB - Experiments on different annular denuder system (ADS) arrangements for sampling nitrous acid (HNO2) and nitric acid (HNO3) gases were conducted in this study to evaluate their sampling artifacts. The evaluation basis is the one that employed one sodium chloride denuder for sampling HNO3 gas and two sodium carbonate (Na2CO3) denuders for sampling HNO2 gas, which is a commonly employed ADS arrangement in many field applications in the United States. A field study was conducted in Hsinchu, Taiwan, and the results indicated that this ADS arrangement may yield over 80% relative errors for HNO3 gas. It also showed that the relative errors for HNO2 gas can be less than 10% as sampled with only one Na2CO3 denuder. This is attributed to the fact that the ambient HNO3 concentration measured in this study was relatively low while the HNO2 concentration was high, as compared to typical concentrations of these two gases measured in the United States. The sampling error of HNO3 gas may be due to high concentrations of N-containing interfering species present in Taiwan's atmosphere. Because the relative sampling errors of HNO3 and HNO2 gases depend mainly on their concentrations in the atmosphere as well as concentrations caused by interfering species, the risk for high error while measuring low HNO2 concentrations by only one Na2CO3 denuder is also possible. As a result, it is suggested that pretests are necessary to evaluate possible sources and degrees of sampling errors before field sampling of HNO2 and HNO3 gases. The sampling errors of these two gases can, therefore, be minimized with a better arrangement of the ADS. PMID- 10680374 TI - Concentration of atmospheric pollutants in the gaseous emissions of medical waste incinerators. AB - The purpose of this paper is to quantify the production of medical waste from a general hospital and to evaluate the atmospheric pollutant concentrations in gaseous emissions associated with its incineration. A 3.8 kg (bed.day)-1 production of medical waste was estimated for 1998; its incineration is related with an ash production of 0.3-0.4 kg (bed.day)-1. The concentrations of atmospheric pollutants were estimated using emission factors, comparing the effluents with and without control of atmospheric pollutants. The calculated concentrations were compared with the emission limits established by Portuguese legislation. The results indicate that, if there is no control of atmospheric pollutants, their concentrations exceed the established limits. This is observed even if correct operation and maintenance procedures are used. The emission concentrations of dioxins are higher than the Portuguese emission limit, which is particularly worrying due to the high toxicity of some of these compounds. Generally, it is possible to reduce pollutant concentrations if appropriate control equipment is used. The conclusions obtained clearly justify the great concern regarding air pollution associated with medical waste incinerators currently operating in Portugal. PMID- 10680375 TI - Sulfur hexafluoride (SF6): global environmental effects and toxic byproduct formation. AB - This work provides information concerning possible global environmental implications and personnel safety aspects that should be considered during the commercial uses of sulfur hexafluoride (SF6). SF6 is an anthropogenically produced compound, mainly used as a gaseous dielectric in gas insulated switchgear power installations. It is a potent greenhouse gas with a high global warming potential, and its concentration in the earth atmosphere is rapidly increasing. During its working cycle, SF6 decomposes under electrical stress, forming toxic byproducts that are a health threat for working personnel in the event of exposure. Several precautions are recommended to avoid personnel exposure to toxic byproducts: oxyfluoride levels or other byproduct concentrations in the operating gas matrix should be traced to predetermine the overall gas toxicity; contaminants should be systematically considered during maintenance, chamber evacuation and system opening process; small SF6 quantities leaking into air or stagnated pollutant concentrations in the operating field should be analyzed and compared to the threshold limit values and permissible exposure levels. New system design rules (i.e., hermetically sealed gas compartments, gas recycling or disposal in the field area) and different handling policies--both during maintenance and final disposal--now should be considered globally to provide for environmental and personnel safety. PMID- 10680376 TI - Predictions galore. PMID- 10680377 TI - Fugi I luting cement. PMID- 10680378 TI - Delegation a disservice? PMID- 10680379 TI - New FDA web site targets online quackery. PMID- 10680380 TI - Feds warn hospitals about drug-resistant infections. PMID- 10680381 TI - Surveys says Americans concerned about diet. PMID- 10680382 TI - Name one change or development in dentistry that you believe the profession will witness in the 21st century. PMID- 10680383 TI - Myofascial face pain. Clinical characteristics of those with regional vs. widespread pain. AB - BACKGROUND: The authors conducted a study to determine whether there are differences in salient clinical characteristics between patients who have both myofascial face pain, or MFP, and comorbid fibromyalgia, or FM, and patients who have MFP but not FM. METHODS: The authors enrolled in the study 162 female subjects who had histories of MFP. In physical examinations at the time of initial consultation, they recorded facial pain signs and symptoms. At the research interview follow-up (seven years post-consultation), participants were screened for a lifetime history of FM and other health problems. In addition, psychiatric interviewers conducted the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Third Edition-Revised, to assess each patient's history of depression and other psychiatric disorders. RESULTS: Of the 162 participants, 38 (23.5 percent) reported a history of FM. At the time of treatment for MFP, both the FM and non-FM groups had similar signs and symptoms of MFP. At the time of the research interview follow-up, participants with FM histories were significantly less likely than those without FM histories to report that they were free of MFP. On recall, those with FM histories reported experiencing more symptoms of MFP. Those with FM histories also were more likely to have had major depression and to report somatization symptoms. Finally, those who had FM more commonly had a history of facial pain's interference with social and occupational functioning and had more severe pain than did those without FM. CONCLUSIONS: Patients who have MFP and a history of widespread pain suggestive of FM are likely to have more persistent and debilitating MFP and to have higher rates of depression and somatization symptoms than those who have no history of widespread pain. PMID- 10680384 TI - Causes of failure among cuspal-coverage amalgam restorations: a clinical survey. AB - BACKGROUND: Investigations of cuspal-coverage amalgam restorations suggest that tooth fracture is the leading cause of failure, while for Class I and II restorations, the leading cause is caries. In this study, the authors evaluated the causes of failure for a large number of cuspal-coverage restorations. METHODS: The causes of failure for 706 cuspal-coverage amalgam restorations were determined through the use of a questionnaire. Dentists from a variety of dental schools; Army, Navy, Air Force, Public Health and Veterans Affairs dental clinics; and private practice were asked to record pertinent information regarding patients and restoration failures from choices provided on a survey form. RESULTS: The survey documented 706 failed restorations. Mandibular first molars accounted for 36.25 percent of all failures. The majority of failures were caused by fractured teeth (24.3 percent), caries (20 percent) and fractured restorations (17.1 percent). Among all of the failed restorations, 82.15 percent were restorable, 9.35 percent were repairable and 8.50 percent were nonrestorable. Among the fractured teeth, 80 percent were restorable, 14.5 percent were nonrestorable and 5.5 percent were repairable. Among the carious teeth, 84 percent were restorable, 8 percent were nonrestorable and 8 percent were repairable. A chi 2 analysis revealed that tooth fracture was more likely to be associated with nonrestorability than either caries (chi 2 = 5.013, P < .05) or restoration fracture (chi 2 = 6.202, P < .05). CONCLUSIONS: The leading cause of failure among the 706 restorations was tooth fracture, which resulted in significantly greater numbers of nonrestorable teeth than either caries or fractured restorations. CLINICAL IMPLICATIONS: Tooth fracture creates a greater risk of nonrestorability than any other cause of failure. Replacement or coverage of fracture-prone cusps is likely to improve the life expectancy of complex amalgam restorations. PMID- 10680385 TI - Dental unit waterline antimicrobial agents' effect on dentin bond strength. AB - BACKGROUND: In response to concerns of bacterial biofilm colonization of dental unit waterlines, a wide range of commercial intermittent and continuous chemical treatments for dental unit waterlines have been developed and marketed. There has been little research on the possible effect of continuous chemical treatment regimens on dentin-bonding agents. The authors evaluate the effect of four proposed antimicrobial agents used in dental unit waterlines on dentin bond strength. METHODS: The authors used a fifth-generation dentin-bonding agent to bond composite cylinders to molar dentin surfaces. They then used selected antimicrobial agents as rinsing agents after conditioning. The composite cylinders were shear tested, and their fracture strengths were compared statistically. RESULTS: All proposed antimicrobial agents reduced dentin bond strength. Proposed waterline treatment regimens of a diluted mouthrinse and chlorhexidine significantly reduced dentin bond strength compared with sodium hypochlorite and citric acid regimens. CONCLUSION: Dental professionals should be aware of potential interactions between dental unit waterline antimicrobial agents and dentin-bonding agents. Further research in this area is warranted, as the clinical implications are uncertain at this time. CLINICAL IMPLICATIONS: Dental unit waterline antimicrobial agents may adversely affect dentin bonding strength. PMID- 10680386 TI - Topical medications for orofacial neuropathic pain: a review. AB - BACKGROUND: The authors discuss the local pharmacotherapy for chronic orofacial neuropathic pain disorders such as neuropathies, neuromas and neuralgias. METHODS: The authors conducted a systematic literature review on this topic. The focus of the review involved the two most commonly applied medications for neuropathic disorders--local anesthetics and capsaicin. Other compounds such as nonsteroidal anti-inflammatory drugs, sympathomimetic agents, anticonvulsants and N-methyl-D-aspartate receptor antagonists also were reviewed. The medication delivery and retention methods appropriate for oral and perioral disease and pain control are described in this article. RESULTS: There are an ever-increasing number of agents that can be used to help patients with neuropathic-based oral and perioral pain problems. Moreover, a clear advancement in the delivery of these medications is the development of a vehicle-carrier agent (pluronic lecithin organogel) that can penetrate the mucosa and cutaneous tissues and carry the active medication with it to the treatment site. The major caveat underlying these treatment strategies is that except for patient testimony and a few studies, there are limited empirical data on the efficacy of most of these new formulations, and additional research is clearly needed. CONCLUSIONS: Because of their rapid onset and low side-effect profile, topical medications offer a distinct advantage over systemic administration for those orofacial disorders that are regional, near the surface and chronic and that demonstrate some response such as pain relief to topical or subcutaneous anesthetics. CLINICAL IMPLICATIONS: Practicing dentists now have some new tools they can use to help manage patients who have a chronic nerve pain disorder in and around the mouth. PMID- 10680387 TI - Repairing iatrogenic root perforations. AB - BACKGROUND: Post preparation is an integral part of restoring endodontically treated teeth in indicated cases. Iatrogenic perforation of the root can result from preparing post space and can severely compromise the prognosis of the tooth. CASE DESCRIPTION: Two years after a patient's maxillary lateral incisor was restored with a post-retained composite resin, he went to a dental school emergency clinic with a chief complaint of soft-tissue swelling adjacent to the tooth. The authors took a periapical radiograph that revealed evidence of a circumscribed radiolucent lesion associated with the distal midroot area and a periapical radiolucency. Based on the radiograph, the authors suspected that the canal preparation for the post and the post placement had perforated the root at the base of the post. CLINICAL IMPLICATIONS: The authors used a combined surgical and orthograde approach with a biocompatible restorative material and a clear, plastic light-transmitting post to repair the iatrogenic perforation. PMID- 10680388 TI - Usefulness of posture training for patients with temporomandibular disorders. AB - BACKGROUND: Many practitioners have found that posture training has a positive impact on temporomandibular, or TMD, symptoms. The authors conducted a study to evaluate its effectiveness. METHODS: Sixty patients with TMD and a primary muscle disorder were randomized into two groups: one group received posture training and TMD self-management instructions while the control group received TMD self management instructions only. Four weeks after the study began, the authors reexamined the subjects for changes in symptoms, pain-free opening and pressure algometer pain thresholds. In addition, pretreatment and posttreatment posture measurements were recorded for subjects in the treatment group. RESULTS: Statistically significant improvement was demonstrated by the modified symptom severity index, maximum pain-free opening and pressure algometer threshold measurements, as well as by the subjects' perceived TMD and neck symptoms. Subjects in the treatment group reported having experienced a mean reduction in TMD and neck symptoms of 41.9 and 38.2 percent, respectively, while subjects in the control group reported a mean reduction in these symptoms of 8.1 and 9.3 percent. Within the treatment group, the authors found significant correlations between improvements in TMD symptoms and improvements in neck symptoms (P < .005) as well as between TMD symptom improvement and the difference between head and shoulder posture measurements at the outset of treatment (P < .05). CONCLUSIONS: Posture training and TMD self-management instructions are significantly more effective than TMD self-management instructions alone for patients with TMD who have a primary muscle disorder. PRACTICE IMPLICATIONS: Patients with TMD who hold their heads farther forward relative to the shoulders have a high probability of experiencing symptom improvement as a result of posture training and being provided with selfmanagement instructions. PMID- 10680389 TI - Treating severe bruxism with botulinum toxin. AB - BACKGROUND: Locally administered botulinum toxin, or BTX, is an effective treatment for various movement disorders. Its usefulness in treating bruxism, however, has not been systematically evaluated. SUBJECTS AND METHODS: The authors studied 18 subjects with severe bruxism and whose mean duration of symptoms was 14.8 +/- 10.0 years (range three-40 years). These subjects audibly ground their teeth and experienced tooth wear and difficulty speaking, swallowing or chewing. Medical or dental procedures had failed to alleviate their symptoms. The authors administered a total of 241 injections of BTX type A, or BTX A, in the subjects' masseter muscles during 123 treatment visits. The mean dose of the BTX A was 61.7 +/- 11.1 mouse units, or MU (range 25-100 MU), per side for the masseter muscles. RESULTS: The mean total duration of response was 19.1 +/- 17.0 weeks (range six 78 weeks), and the mean peak effect on a scale of 0 to 4, in which 4 is equal to total abolishment of grinding, was 3.4 +/- 0.9. Only one subject (5.6 percent) reported having experienced dysphagia with BTX A. CONCLUSION: The results of this study suggest that BTX administered by skilled practitioners is a safe and effective treatment for people with severe bruxism, particularly those with associated movement disorders. It should be considered only for those patients refractory to conventional therapy. Future placebo-controlled studies may be useful in further evaluating the potential of BTX in the treatment of bruxism. PMID- 10680390 TI - Limiting sensitivity after quadrant scaling and root planing. PMID- 10680391 TI - Trends in total caries experience: permanent and primary teeth. AB - BACKGROUND: This article is the last in a series of three that focuses on recent changes in the caries status of children aged 18 years or younger in the United States. METHODS: This study is based on analyses of data regarding total carious (treated and untreated) permanent and primary teeth among children 6 to 18 years old and children 2 to 10 years old from the first and third National Health and Nutrition Examination Surveys, or NHANES I and NHANES III. The NHANES is periodically conducted by the National Center for Health Statistics of the Centers for Disease Control and Prevention. RESULTS: The cumulative number of carious permanent teeth, both treated and untreated, among 6- to 18-year-olds decreased 57.2 percent, from 4.44, as measured in NHANES I, to 1.90, as measured in NHANES III. The cumulative number of carious primary teeth, both treated and untreated, among 2- to 10-year-olds decreased 39.7 percent, from 2.29, as measured in NHANES I, to 1.38, as measured in NHANES III. CONCLUSIONS: Since the 1970s, the cumulative number of carious permanent and primary teeth, both treated and untreated, has declined substantially among children in the United States. PRACTICE IMPLICATIONS: Effective prevention has reduced caries in children. As a result, dental practice will be more focused on maintaining intact dentitions than on repairing teeth damaged by disease. PMID- 10680392 TI - Treating bruxism and clenching. PMID- 10680393 TI - Another approach to learning about health disparities: working toward individualized therapy. PMID- 10680394 TI - The war on fraud and its effect on dentistry. AB - The fraudulent actions of a disreputable few people in the health care industry have caused enormous losses to government and private health care payers. Fraud can take a number of forms, but usually is based on some form of deceit. In response, law enforcement efforts have been aided by a number of recent anti fraud statutes and significant new resources. The challenge to dentistry is to ensure that legitimate efforts to fight fraud do not unduly burden practitioners, the vast majority of whom are honest. PMID- 10680395 TI - Ask the expert. How much overhead is too much? PMID- 10680396 TI - Holding HMOs responsible. PMID- 10680397 TI - The woman physician in the year 2000. PMID- 10680398 TI - American women physicians in 2000: a history in progress. AB - This article surveys major trends in the history of women physicians in American medicine during the 20th century, noting especially factors that have elicited renewed and increasingly public attention during the past two decades. These include the challenges of achieving greater professional visibility while also balancing family and career, of sustaining women physicians' legacy of commitment to women's health and primary care medicine without reinforcing the traditional stereotype that these are the specialties "best suited" to women doctors, and of addressing the need for more ethnic and racial diversity in the medical profession. Other recent developments include the leveling off of the number of women entering medical school and the increasing tendency of both men and women physicians to practice as employees. PMID- 10680399 TI - Women in academic medicine. AB - Women now constitute 43% of US medical students, 37% of residents, and 27% of full-time medical school faculty. Less than 11% of women faculty are full professors, however, compared to 31% of men, and these proportions haven't changed in more than 15 years. Since the proportion of women reaching the top ranks remains relatively low, the pool of women available for leadership positions in academic medicine is still small. This review article first summarizes recent data on women's representation in academic medicine and then discusses why they are not succeeding at the same pace as men. Reasons include a complex combination of women's choices, sexism, cultural stereotypes, constraints in combining family responsibilities with professional opportunities, and lack of effective mentoring. Multiple approaches are required to overcome these "cumulative disadvantages," among them improving the gender climate at academic medical centers; the mentoring of women faculty, residents, and students; and skill-building opportunities for women. PMID- 10680400 TI - What impact have women physicians had on women's health? AB - The proportion of women in medicine in the United States is approaching that of men, but women physicians are still the minority in positions of power. Research has shown that women physicians order more preventive tests for women patients, are more attuned to patients' psychosocial needs, and have developed more patient oriented communication styles than men physicians. Recent organized efforts of women physicians have brought attention to many gender gaps in medical research and practice. If more women move into policy-making positions, especially in education and research, their gender-sensitive perspective could permanently influence the profession as a whole. PMID- 10680401 TI - Physician unions: organizing women in the year 2000. AB - Interest in physician unions is growing, but surprisingly little has been written about whether union membership addresses the particular needs and interests of women physicians. We begin by looking at the history of physician unions in the United States and then examine physician unions today, and how labor laws influence union membership of physicians. The third section looks at why women join unions, whether these reasons hold true for women physicians, and what role women are playing in physician unions. Finally, we give examples of union responses to gender discrimination and such issues as maternity leave, salary inequities, sexual harassment, and promotions. Since women are prominent as leaders in physician unions, these unions seem to be responsive to the needs of their women members. PMID- 10680402 TI - The subspecialty training, practice type, and geographical distribution of recently trained ophthalmologists: a study of male and female physicians. AB - OBJECTIVE: To characterize the distribution of male and female ophthalmologists with regard to practice type, subspecialty training, rural-urban distribution, and regional distribution. METHODS: Ophthalmology Matching Program files containing the records of residents who began their second year at accredited programs between 1986 and 1990 (inclusive), were compared to membership files of the American Academy of Ophthalmology. Practice locations for each individual were classified according to region, stage, and Rural-Urban Continuum County Code, as defined by the US Department of Agriculture. RESULTS: This cohort comprised 2,494 individuals, 77.1% (1922) of whom were male and 22.9% (572) of whom were female. Group practice was most common (55.9% for women and 61.3% for men). More women were in salaried positions associated with health maintenance organizations (p = 0.006) and academic settings (p < 0.001) than were men. Notable differences in subspecialty choice were restricted to pediatric ophthalmology, chosen three times more frequently by women, and vitreoretinal diseases/surgery, chosen twice as often by men. Only 5.6% of women selected nonmetropolitan practice locales compared to approximately twice that percentage of men. The Middle Atlantic and New England regions attracted more women, while the South Atlantic attracted more men. PMID- 10680403 TI - Medical faculty's views and experiences of parental leave: a collaborative study by the Gender Issues Committee, Council of Ontario Faculties of Medicine. AB - OBJECTIVES: To examine medical faculty's actual and ideal parental leave arrangements with the aim of informing policy decisions. Leave lengths, effect on career, financial arrangements, and availability of temporary replacements were explored. METHODS: All medical faculty (6387) in Ontario, Canada were surveyed by mail and asked about parental leave experiences since 1990. Responses of men and women were compared as were those of leave takers and the entire group. RESULTS: Thirty-two percent (n = 996) of the 3107 respondents were women and 68% (n = 2067) were men. Ninety-eight percent (n = 317) of new mothers had taken maternity leave, while only 21% (n = 159) of new fathers had. Both paid and unpaid leave was generally shorter than that allowed by law or identified as ideal. Parental leave had a somewhat negative effect on the careers of all faculty. Women were more worried than men about the effect of their absence on colleagues' work and more generous with ideal leave length and funding. Temporary replacement of leave takers was central to an effective leave policy. CONCLUSIONS: Institutional and academic culture may cause new parents to take suboptimal leave despite legislation allowing more. A change in the work environment is required for medicine to offer its teachers what it teaches--that infants benefit from nurturing, nursing, and stability early in life. PMID- 10680404 TI - Continuing medical education habits of US women physicians. AB - OBJECTIVES: To quantify the time women physicians spend on continuing medical education (CME) in total and by type of activity and to assess the impact of various demographic and professional characteristics on CME practices. METHODS: We analyzed CME habits among 4501 female physicians (a 59% response rate) using a 1994 national questionnaire-based survey, the Women Physicians' Health Study. RESULTS: US women physicians reported spending a monthly mean of 12.5 hours on CME, including 5.1 hours reading medical journals, 3.1 reading medical textbooks, 3.7 attending live CME lectures, 0.7 listening to CME audio tapes, and 0.4 watching medical television or videos. They also spent 0.9 hours per month learning from lay health media. Physicians with subspecialty training, medical school employment, and a non-US birthplace reported significantly (p < 0.05) more CME hours; age, ethnicity, region, specialty type, practice locale, career satisfaction, and board certification did not significantly predict reported CME hours. CONCLUSIONS: US women physicians reported spending an average of one-half hour each work day on CME, including about one hour per week reading medical journals, their most commonly reported CME activity. PMID- 10680405 TI - Differences in the preparation and practice of male and female physicians from combined baccalaureate--MD degree programs. AB - OBJECTIVE: To account for age and premedical education in determining whether or not men and women medical school graduates differ significantly in opinions of their medical school preparation, in professional activities, and in personal qualities and values. METHODS: 727 graduates (1983-1987) of seven combined baccalaureate-MD degree programs were surveyed. Men's and women's responses were compared. RESULTS: Women graduates were more likely than men to work less than full time and to report less preparation in the basic sciences, less scholarly activity, and more concern about psychosocial issues. CONCLUSION: Differences between men and women graduates have persisted despite women's increased access to medical school. Even a special curriculum does not moderate these differences. We call on academic medicine to value humanism in health care, not just economics. PMID- 10680406 TI - The gender composition of the medical profession in Mexico: implications for employment patterns and physician labor supply. AB - The gender composition of the medical profession is changing rapidly in many parts of the world, including Mexico. We analyze cross-sectional and longitudinal data on sex differences in physician employment from household employment surveys. The results suggest that Mexico is a particularly interesting example of the feminization of physician employment. Female enrollment in medical school increased from 11% in 1970 to about 50% in 1998. The increased participation of women in medicine seems to be accompanied by differences in employment patterns that could generate significant reductions in the total supply of physician hours of service. Women physicians are unemployed at a much higher rate than men and hence account for half of underused physician human capital. The results suggest that improved educational opportunities do not translate automatically into equal employment opportunities. PMID- 10680407 TI - Egyptian medical women, past and present. AB - In ancient Egypt, at least one woman carried the title of physician. University education for women started in 1930. Today, women are practicing in all disciplines and are effective in health care. Egyptian medical women represent 35% to 45% of the staff of faculties of medicine and about one-third of all medical graduates. They have contributed to the improvement of health, particularly in maternal and child health, and are role models for young girls in rural areas. PMID- 10680408 TI - Why single payer? Why now? AB - US health care costs are the highest in the world and are again rising. A reopening of debate on health care reform is imminent. More than 44 million Americans have no health insurance, an increase of 11 million people since 1989. Although women have been slightly more likely to have health insurance than men, recent declines in Medicaid enrollment resulting from welfare reform are eroding this slim advantage. Being uninsured is associated with compromised access to primary care and an increased risk of dying. At least 29 million Americans are underinsured; although they have some insurance, they would nonetheless be bankrupted by a major illness. A single-payer national health insurance system would cover all Americans in a non-profit, tax-funded system similar to Social Security. It would simplify health administration, saving at least $100 billion annually on paperwork and redirecting that money to patient care. PMID- 10680409 TI - Prevention works for women: women's health at the CDC. AB - Actively protecting America's health and safety, the Centers for Disease Control and Prevention (CDC) focuses on four priorities: 1) strengthening science for public health action, 2) collaborating with health care partners for prevention, 3) promoting healthy living at every stage of life, and 4) promoting health globally. The CDC plays a critical role in promoting the health and safety of women across the lifespan, through programs on human immunodeficiency virus infection, injury, contraceptive safety and efficacy, adolescent health, smoking, breast and cervical cancer, cardiovascular disease, and reproductive health. PMID- 10680410 TI - Randomized community trial of a breast self-examination skills-building intervention for inner-city African-American women. AB - OBJECTIVE: Breast cancer is the most common cancer in women, and African-American women are less likely to detect breast cancer at its early stages. Few controlled trials of interventions to increase breast self-examination (BSE) among low income minority women have been conducted. Our objective was to test a small group workshop intended to build BSE skills and promote BSE among low-income African-American women. METHODS: A randomized community field trial tested a BSE skills-building workshop based on social cognitive theory of behavior change compared to a matched sexually transmitted diseases prevention workshop, with one and three-month follow-ups for assessing increased practice of BSE. RESULTS: Women who did not regularly practice BSE and participated in a BSE skills building workshop were more likely to practice BSE than women in the comparison intervention at the one- (OR = 3.5, p = 0.04) and three-month follow-ups (OR = 4.9, p = 0.01). These results were not related to risk perceptions heightened by the intervention. Across conditions and controlling for covariates in a multivariate model, performing regular BSE was most closely associated with having received any formal BSE instruction. CONCLUSION: BSE skills building can effectively increase use of BSE among low-income African-American women who face multiple and competing health risks. The small-group experience is an important element in fostering norms for practicing BSE and enhancing BSE practices. PMID- 10680411 TI - "The speed of blur...". PMID- 10680412 TI - Optometry's research profile: we are on the way up; how high can we go? PMID- 10680413 TI - The changing faces of optometry. PMID- 10680414 TI - Competency-based optics instruction. AB - BACKGROUND: The Competency-Based Instruction (CBI) system has been used to teach physics for more than 20 years in the Michigan State University Physics Department. In this approach, traditional lectures have been replaced by a learning environment that contains a variety of instructional aids, including written materials, computer-assisted instruction, and interactions with a consultant. The CBI system allows students to adjust their pace through the course, moving nearly as quickly as they are able, with constant feedback to the student on his or her progress. METHODS: I have used an adapted version of the CBI system for use in the Physical Optics and Photometry course of the University of Missouri-St. Louis, School of Optometry for four years. This article will describe the mechanics of the system and discuss experiences with it. PMID- 10680415 TI - Pacific's experience with Web-based instruction: bats in the belfry or Webs in the classroom? AB - BACKGROUND: There is a revolution underway in education that involves a shift from the traditional lecture style of information presentation toward a more active style of learning. Many educators now believe that students must actively participate in the learning process for information to be truly understood and retained. Coincident with this revolution, there is an increased understanding that the body of knowledge in most professions has become too large to retain in the brains of individuals. These concepts have opened the door to innovative, computer-based educational techniques. METHODS: Pacific University College of Optometry has used a variety of Web-based educational methodologies in selected classes. These have ranged from shifting classes totally to the Web, to using the Web for pre-class preparation. RESULTS: Initial student acceptance of Web-based courses has been good. However, an undesirable trend has emerged--the tendency for students to print out Web-presented material rather than reading from the computer screen. Other concerns with shifting material from lecture to the Web include peer and administrator acceptance of this teaching style and issues associated with evaluation of professors who no longer give stand-up lectures. CONCLUSIONS: Changes in educational theory are making their way into the Schools and Colleges of Optometry. These changes will place greater reliance on active learning, reduced student memorization, and increased use of computers as information storage and retrieval devices. This will change the fundamental way in which students are educated, and this, in turn, will change the way in which future doctors, patients, and computers interact in clinical settings. PMID- 10680416 TI - Ocular toxicity of systemic medications: a case series. AB - BACKGROUND: There are many visually threatening conditions that may result from long-term use of systemic medications. Many of these adverse side effects can be greatly reduced or prevented with close monitoring of patients. In view of current knowledge, updated clinical guidelines for appropriate monitoring of ocular toxicity from systemic medications need to be developed for the eye care practitioner. CASE REVIEW: There have been many reports of ocular toxicity from isoniazid, thioridazine, steroids, and amiodarone therapy. Clinical cases illustrating possible adverse ocular side effects are presented, which include INH-induced optic neuropathy, phenothiazine-induced retinopathy, steroid-induced glaucoma, and vortex epitheliopathy secondary to amiodarone. CONCLUSION: Optometrists should be aware of the potential for ocular side effects from systemic medications. Eye care guidelines for monitoring ocular side effects from thioridazine, INH, steroids, and amiodarone use are suggested. PMID- 10680417 TI - Iowa's pediatric low-vision services. AB - BACKGROUND: Reports in the literature concerning best practice for the evaluation and management of children with visual impairments are limited, with a resulting lack of information concerning the potential for optimizing vision to enhance general development and assist with the educational needs of this population. METHOD: The development of a multidisciplinary approach to provide low-vision services for children with visual impairment has occurred over the past 18 years in Iowa. In that time, 1,348 children from around the state of Iowa have been evaluated through an itinerant low-vision service program, coordinated by the Iowa Braille School. RESULTS: A low-vision clinic model--designed to provide services (primarily) for academic students--was not meeting the needs of the pediatric low-vision population in the state. After a statewide review of the program, changes were made that have resulted in low-vision services being provided to a greater and more diverse number of students. The roles of the various members of the multidisciplinary team will be reviewed. Changes in large print orders and use by special education teachers in the state as a direct result of the low-vision services will also be discussed. CONCLUSION: Ongoing, comprehensive multidisciplinary low-vision services--including optometric low vision care as a key component--are necessary to help children with visual impairments meet their educational, vocational, and avocational needs. With ongoing low-vision services, unnecessary costs such as those associated with large-print materials can be reduced, thereby creating significant savings to local, state, and federal special educational services. PMID- 10680418 TI - A comparison of Fluoracaine and Fluorox on corneal epithelial cell desquamation after Goldmann Applanation Tonometry. AB - BACKGROUND: The purpose of this study was to quality the frequency and amount of corneal desquamation from a sodium fluorescein/proparacaine combination (Fluoracaine) as compared with sodium fluorescein/benoxinate combination ophthalmic solution (Fluorox) after Goldmann Applanation Tonometry. METHODS: One drop of Fluoracaine was randomly instilled into one eye and one drop of Fluorox was instilled into the opposite eye of the same patient. Intraocular pressures (IOPs) by GAT and tear break-up times (TBUTs) were taken. Corneal stinging was compared. Corneal integrity by Cornea and Contact Lens Research Unit (CCLRU) standards was evaluated at 0, 3, 7, 10, 15, and 20 minutes after instillation of the ophthalmic solutions. RESULTS: Sixty eyes of 30 patients were observed Forty seven percent of the patients reported Fluorox to string more than Fluoracaine; 23% of the patients reported that Fluoracaine stings more than Fluorox; and 30% the patients reported no difference. Average TBUTs were 6.87 and 7.17 seconds with Fluoracaine and Fluorox, respectively. Fluoracaine produced micro- and macropunctate keratitis of the superficial epithelium in 31% to 45% of the cornea. Fluorox caused superficial micropunctate keratitis in about 16% to 30% of the cornea. At 20 minutes, all eyes with Fluoracaine and all eyes but one with Fluorox had corneal desquamation. CONCLUSIONS: Fluoracaine causes marginally less stinging--however, clinically and statistically more corneal desquamation--than Fluorox after GAT. Corneal integrity after use of Fluoracaine should be evaluated even 20 minutes after GAT procedures for corneal disruption. PMID- 10680419 TI - Internet research: improving traditional community analysis before launching a practice. AB - Optometric practice management experts have always recommended that optometrists thoroughly research the communities in which they are considering practicing. Until the Internet came along, demographic research was possible but often daunting. Today, say these authors, it's becoming quite a bit easier ... and they show us how. PMID- 10680420 TI - Is an online discount broker right for you? AB - Let's face it ... you will eventually do all your investing online, if you're not doing so already. But how can you distinguish between the many brokerage choices? These authors offer their insight into finding the best online broker at the most reasonable cost. PMID- 10680421 TI - Computing & technology. PMID- 10680422 TI - The drug war. PMID- 10680423 TI - The costly quest for certainty. PMID- 10680424 TI - Not insured, and not worried. PMID- 10680425 TI - The cost of coverage. PMID- 10680426 TI - Health care decisions and the community. PMID- 10680427 TI - Community forums bring together Minnesota's health care players. PMID- 10680428 TI - Is the experimental treatment exclusion used appropriately? PMID- 10680429 TI - Pediatrics: a century of caring. PMID- 10680430 TI - MMA reviews prescription drug plans. PMID- 10680431 TI - Minnesota's Medicare PRO focuses on quality improvement. PMID- 10680432 TI - Diagnosis and management of carotid stenosis: a review. AB - Since its introduction in the 1950s, carotid endarterectomy has become one of the most frequently performed operations in the United States. The tremendous appeal of a procedure that decreases the risk of stroke, coupled with the large number of individuals in the general population with carotid stenosis, has contributed to its popularity. To provide optimal patient care, the practicing physician must have a firm understanding of the proper evaluation and management of carotid stenosis. Nevertheless, because of the large number of clinical trials performed over the last decade addressing the treatment of stroke and carotid endarterectomy, the care of patients with carotid stenosis remains a frequently misunderstood topic. This review summarizes the current evaluation and treatment options for carotid stenosis and provides a rational management algorithm for this prevalent disease process. PMID- 10680433 TI - Minnesota surveillance for unexplained deaths and critical illnesses of possible infectious cause. PMID- 10680434 TI - Itraconazole trough concentrations in antifungal prophylaxis with six different dosing regimens using hydroxypropyl-beta-cyclodextrin oral solution or coated pellet capsules. AB - We have previously shown that a trough concentration of at least 500 ng ml-1 itraconazole is necessary for an effective antifungal prophylaxis in neutropenic patients. Since the bioavailability of itraconazole is reduced in these patients, a satisfactory dosing regimen remains to be defined. In this study, six dosing regimens with itraconazole capsules 400, 600 or 800 mg day-1, itraconazole solution 400 mg day-1 (additional loading dose: 400 mg day-1 solution for 2 days), 800 mg day-1 or 400 mg day-1 (additional loading dose: 800 mg day-1 capsules for 7 days, s/c1200) were compared during 160 courses of myelosuppressive chemotherapy in 123 patients with acute leukaemia. After the first week, patients taking 800 mg day-1 or 400 mg day-1 (s/c1200) itraconazole solution achieved significantly higher trough concentrations (high-performance liquid chromatography) than patients in other groups (P < 0.05) and 87 and 100%, respectively, of these had concentrations > 500 ng ml-1. Contrary to a dose of 400 mg day-1, a dose of 800 mg day-1 itraconazole solution induced severe nausea and vomiting in 46% of the patients. We conclude that 400 mg day-1 itraconazole solution with a loading dose of 800 mg day-1 capsules for 7 days resulted in sufficient trough concentrations from the first week onwards and appears to be suitable for antifungal prophylaxis in neutropenic patients. PMID- 10680435 TI - Killer activity at different pHs against Cryptococcus neoformans var. neoformans serotype A by environmental yeast isolates. AB - Yeast isolates that share the same habitats as Cryptococcus neoformans var. neoformans serotype A in a restricted Mediterranean area were assayed in order to verify their killer activity against Cr. neoformans strains isolated from clinical and environmental sources. Many of the environmental yeast isolates expressed the killer phenomenon against the assayed strains of Cr. neoformans. Two species of Candida: Candida parapsilosis and Candida famata, and Pichia carsonii, were the most active killers at pH 4.6, 5.0 and 5.6 levels encountered in pigeon and canary guanos. Killer activity by C. parapsilosis is reported for the first time. The authors hypothesized that the killer phenomenon exerted by yeast species with heavy killer activity against Cr. neoformans would lend themselves for use as biological control agents against sensitive strains of Cr. neoformans when directly inoculated into the habitats of Cr. neoformans. PMID- 10680436 TI - Dematiaceous fungal pathogens: analysis of ribosomal DNA gene polymorphism by polymerase chain reaction-restriction fragment length polymorphism. AB - Restriction fragment length polymorphisms (RFLP) of ribosomal gene small subunit (SSU rDNA) and internal transcribed spacer (ITS) regions was examined in 12 isolates of dematiaceous agents of chromoblastomycosis and phaeohyphomycosis. The amplicon length of the fragment ITS1-ITS4, comprising the 5.8 rDNA and ITS1-ITS2 spacers, ranged in size from 620 to 690 bp. This result indicated a polymorphism of size in this region. Additionally the RFLP profiles showed a high degree of inter- and intra-specific variability. In contrast, the SSU rDNA amplification, using NS1-NS2 primers, originated a fragment of approximately 570 bp and its restriction profile proved to be well conserved among the species studied and was clustered into only two genetically heterogeneous groups, the first one formed by Fonsecaea pedrosoi and Fonsecaea compacta and the second one formed by Cladophialophora (Cladosporium) carrionii, Cladophialophora (Xylohypha) bantiana, Phialophora verrucosa and Rhinocladiella species. PMID- 10680437 TI - Isolation of total RNA from dermatophytes. AB - We report a method for the preparation of total RNA from the anthropophilic dermatophyte Trichophyton rubrum. To generate large quantities of mycelia, the fungus was grown in liquid culture medium. The harvested mycelial mass was ground to a fine powder in liquid nitrogen and homogenized in guanidine isothiocyanate buffer followed by ultracentrifugation of the obtained suspension through a caesium chloride gradient. Analysis of the prepared RNA showed two prominent ribosomal RNA (rRNA) bands of about 3.36 and 1.82 kb. Northern blot hybridization with a beta-actin cDNA confirmed the high quality of the fungal mRNA. Successful isolation of RNA from two other dermatophyte species, namely Trichophyton mentagrophytes and Microsporum canis, demonstrated the general applicability of the described procedure. PMID- 10680438 TI - Bacterial flora accompanying Candida yeasts in clinical specimens. AB - From 1986 to 1988, in the prefluconazole era, 67,765 clinical specimens from the Gottingen University Hospital were investigated for bacteria and fungi in our institution. Oral and throat swabs, respiratory secretions, gastric juices, faeces, urine, genital swabs, blood, wound secretions and skin swabs were analysed for yeast-like fungi, and opportunistic or pathogenic bacteria. A total of 5195 specimens (7.7%) yielded Candida spp. alone or in combination with bacteria (fungal (F-) group) and 62,570 specimens yielded bacteria only or remained sterile (non-fungal group, N-group). Elevated rates of accompanying bacteria were detected with Candida spp. colonizing blood, urine, and skin. Among the dominant bacterial isolates, the distribution of staphylococci and enterococci did not reflect a distinct association pattern. Among the enterobacterial isolates from patients in intensive care, colonization patterns of the throat, gastric juices, and faeces reflected the use of a selective decontamination of the digestive tract (SDD). A statistically significant association between Candida and enterobacteria of the genus Enterobacter which was unaffected by SDD, was observed throughout this study. Such an association pattern was also observed, to a lesser extent, with the related genera Klebsiella and Serratia, but not with Escherichia coli. PMID- 10680439 TI - In vitro susceptibility of yeasts for fluconazole and itraconazole. Evaluation of a microdilution test. AB - In vitro susceptibilities were determined for a total of 159 clinical isolates and 12 reference strains of yeasts belonging to different Candida species including 94 Candida albicans strains, and further genera such as Cryptococcus, Trichosporon, Geotrichum and Saccharomyces. Minimum inhibitory concentration (MIC) values for fluconazole and itraconazole were assessed using a microdilution technique with the semisynthetic high resolution (HR) medium supplemented with glucose and asparagine but without sodium hydrogen carbonate (pH 7.0), according to a proposal of the working group 'Clinical Mycology' of the German Speaking Mycological Society. Fluconazole MIC values for C. albicans were between 0.125 and > or = 128 micrograms ml-1. Thus, the median of 1 microgram ml-1 showed that the overall fluconazole susceptibility was good. As expected, Candida krusei (seven strains) exhibited diminished in vitro susceptibility with MIC values for fluconazole of 8 to 128 micrograms ml-1 with a median of 64 micrograms ml-1. Some Candida kefyr strains seemed to be less susceptible against fluconazole which was indicated by a MIC90 of 64 micrograms ml-1. Surprisingly, no Candida glabrata isolate exhibited a MIC value greater than 16 micrograms ml-1. Other Candida species, Trichosporon cutaneum, Geotrichum candidum and Saccharomyces cerevisiae showed low MICs to fluconazole. In vitro susceptibility testing of itraconazole revealed that all Candida species except C. albicans, but also Trichosporon cutaneum, Geotrichum candidum, and Saccharomyces cerevisiae exhibited acceptable low MIC values against itraconazole (0.03-2 micrograms ml-1). Their MIC90 values for itraconazole were in the close range between 0.125 and 2 micrograms ml-1. MIC values between 0.125 and 2 micrograms ml-1 were obtained, even for C. krusei strains. On the other hand, the range of C. albicans MICs was between 0.0125 and > or = 16 micrograms ml-1 with MIC50 and MIC90 values of 0.125 and > or = 16 micrograms ml-1, respectively, indicating that a considerable number of yeast strains have high MICs. The comparative evaluation of different experimental conditions revealed that there exists a marked influence both of inoculum size and incubation time on the results of susceptibility testing. Therefore, for routine usage 10(2) CFU ml-1 and 18-24 h incubation time for this microdilution method with HR medium are recommended. PMID- 10680440 TI - In vitro antifungal susceptibility of clinical isolates of Candida spp. from hospitalized patients. AB - A total of 122 Candida spp. strains, isolated from a group of 100 patients hospitalized in the Santa Casa de Misericordia of Belo Horizonte were assayed for in vitro susceptibility to amphotericin B, fluconazole, itraconazole, ketoconazole and flucytosine using a microbroth technique proposed by the National Committee for Clinical Laboratory Standards. In this study large variations were observed among minimum inhibitory concentration values depending on the species tested. The statistical analysis (Kruskal-Wallis test) showed that itraconazole and flucytosine were the more efficient antifungal drugs for most of species, and amphotericin B and fluconazole were the least efficient. PMID- 10680441 TI - The influence of alcohol disinfection of nail samples in the laboratory. AB - Samples of two hundred finger- and toenails cultured after a 70% alcohol disinfection for 2 min were compared with non-disinfected samples cultured on a classical way. Alcohol treatment caused a significant decrease in isolation of yeast and moulds. Both organisms were cultivated at 25.5 and 31%, respectively, when not treated with alcohol, whereas the isolation rate dropped to 10 and 17.5%, respectively, after alcohol treatment. Isolation of dermatophytes decreased from 27.5 to 23.5% when alcohol disinfection was used. In conclusion, although alcohol disinfection of nail samples in the laboratory effectively decreases the isolation rate of unwanted contaminants on Sabouraud glucose agar without cycloheximide, it hampers the isolation of dermatophytes and pathogenic yeasts on cycloheximide-containing media. PMID- 10680442 TI - Topical treatment of dermatophytosis and cutaneous candidosis with flutrimazole 1% cream: double-blind, randomized comparative trial with ketoconazole 2% cream. AB - In a double-blind, randomized study the efficacy and tolerance of flutrimazole 1% cream were compared with ketoconazole 2% cream, applied once daily for 4 weeks, in 60 patients with culturally proven dermatophytosis (47 patients) or cutaneous candidosis (13 patients). Both groups of patients and distribution of target lesions were similar. The sum of clinical scores had an even distribution in both groups at the end of treatment. The proportion of patients with negative microscopy and culture after 4 weeks of treatment was 70% in the flutrimazole group and 53% in the ketoconazole group; seven ketoconazole-treated patients (23%) compared with two flutrimazole-treated patients (6.6%) were asymptomatic carriers (clinically cured with positive cultures) at the end of treatment. At the assessment 6 weeks after the end of therapy the percentages of flutrimazole- and ketoconazole-treated patients with negative mycology were 57 and 70%, respectively. There were one relapse (3.3%) in the ketoconazole group and four (13.3%) in the flutrimazole group. One patient treated with ketoconazole (3%) had a premature termination due to adverse events attributable to the medication. The results of this study show that flutrimazole 1% cream is as effective and safe as ketoconazole 2% cream for Candida and dermatophyte skin infections. PMID- 10680443 TI - Epidemiology of dermatomycoses of humans in central Poland. Part IV. Onychomycosis due to dermatophytes. AB - The total number of dermatophytoses (7393) included 1567 (21.2%) cases of onychomycosis. Etiological factors in descending order were: Trichophyton rubrum (58.8%), Trichophyton mentagrophytes var. granulosum (26.2%), T. mentagrophytes var. quinckeanum, T. mentagrophytes var. interdigitale (5.1%), Trichophyton tonsurans (4.1%), Trichophyton violaceum (1.5%), Trichophyton spec. (1.1%). An increase in the incidence was noted between 1994 and 1996. At present, onychomycosis is third by incidence among all clinical forms of dermatophyte infections of skin and skin appendages in the Lodz region. PMID- 10680444 TI - Epidemiology of dermatomycoses of humans in central Poland. Part V. Tinea corporis. AB - The total number of dermatophytoses (7393) included 2204 (29.8%) cases of tinea glabrosa. Etiological factors in descending order were: Microsporum canis (23.5%), Trichophyton mentagrophytes var. granulosum (21.6%), Trichophyton rubrum (17.8%), Trichophyton tonsurans (10.4%), Epidermophyton floccosum (7.7%), T. mentagrophytes var. quinckeanum (6.0%), Microsporum gypseum (5.3%), Trichophyton violaceum (3.7%), T. mentagrophytes var. interdigitale (2.3%), Microsporum equinum (0.7%), Trichophyton verrucosum (0.4%), Trichophyton spec. (0.4%), Microsporum cookei (0.14%). At present tinea glabrosa is dominant among all clinical forms of dermatophyte infections of skin and skin appendages in the Lodz region. PMID- 10680445 TI - Antifungal activity of plant extracts against dermatophytes. AB - The aqueous extracts (15 micrograms ml-1 medium) of 22 plants used in folkloric medicine in Palestine were investigated for their antifungal activity and minimum inhibitory concentrations (MICs) against nine isolates of Microsporum canis, Trichophyton mentagrophytes and Trichophyton violaceum. The extract of the different plant species reduced colony growth of the three dermatophytes by 36 to 100% compared with the control treatment. Antimycotic activity of the extract against the three dermatophytes varied significantly (P < 0.05) between test plants. Extracts of Capparis spinosa and Juglans regia completely prevented growth of M. canis and T. violaceum. The most active extracts (90-100% inhibition) were those of Anagallis arvensis, C. spinosa, J. regia, Pistacia lentiscus and Ruta chalapensis against M. canis; Inula viscosa, J. regia and P. lentiscus against T. mentagrophytes; and Asphodelus luteus, A. arvensis, C. spinosa, Clematis cirrhosa, I. viscosa, J. regia, P. lentiscus, Plumbago europea, Ruscus aculeatus, Retema raetam and Salvia fruticosa against T. violaceum. The MICs of these most active plants ranged from 0.6 to 40 micrograms ml-1. The three dermatophytes differed significantly with regard to their susceptibility to plant extracts. Trichophyton violaceum was the most susceptible being completely inhibited by 50% of the extracts followed by M. canis and T. mentagrophytes which were completely inhibited by only 23 and 14% of the extracts, respectively. PMID- 10680446 TI - Comments on Malassezia species from dogs and cats. PMID- 10680447 TI - Case report. Localized pulmonary zygomycosis without pre-existing immunocompromised status. AB - Pulmonary zygomycosis rarely occurs without pre-existing immunocompromised disease. A 72-year-old male was found to have a nodular shadow (3 cm x 4 cm) in the right S8 and S9 on a chest X-ray. Right lower lobectomy was performed and histological examination of the resected material demonstrated pulmonary zygomycosis. Hyphae stained positively not only with Grocott-Gomori methenamine silver staining, but also with an anti-Rhizopus oryzae polyclonal antibody. PMID- 10680448 TI - Case reports. Invasive pulmonary aspergillosis in non-neutropenic patients treated with liposomal amphotericin B. AB - We report two cases of invasive pulmonary aspergillosis due to Aspergillus flavus in one patient who with chronic nephritis and to A. fumigatus in another with malignant lymphoma. After receiving intravenous liposomal amphotericin B therapy for 31 and 35 days, respectively, the patients were cured and did not experience any severe adverse effects. PMID- 10680449 TI - Case report. Folliculitis barbae caused by Candida albicans. AB - Folliculitis barbae candidomycetica is a very rare disease and in the scientific literature this manifestation of candidosis is rarely described. In most cases, a connection with predisposing factors is suspected, although in the cases cited this connection was not frequently proved. Furthermore, in our patient who was suffering from a folliculitis candidomycetica no predisposing factors or illnesses were apparent. For this reason, the importance of such aetiological factors of Candida folliculitis must be critically discussed. Effective drugs for use in treatment are fluconazole, itraconazole and ketoconazole. PMID- 10680450 TI - [Insulin-like growth factor-I: pathophysiologic and clinical aspects]. AB - The paper reviews the factors which affect synthesis and metabolic activity of the insulin-like growth factor-I (IGF-I), its receptor and binding proteins as age, nutritional state, hormonal state and liver function. Attention is also focused on the role of IGF-I in pathogenesis of osteoporosis and neoplastic diseases. PMID- 10680451 TI - [Evaluation of selected HLA DRB1 gene alleles as genetic markers of type I diabetes in the population of northeastern Poland]. AB - Insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease and both environmental and genetic factors play a role in its pathogenesis. The second generation screen of human genome has confirmed that diabetes mellitus type 1 is a polygenic disease. The identification of the alleles with the highest association with IDDM in the different populations gives the opportunity for better evaluation of the probability of IDDM development in subjects at risk (i.e. first degree relatives of IDDM patients with autoantibodies against B cells). The aim of the present study was to estimate the frequency of chosen HLA DRB1 gene alleles in patients with insulin-dependent diabetes mellitus and their first degree relatives in comparison with the healthy population in the north eastern region of Poland. The highest predisposition to IDDM in the population of the north-eastern region of Poland was associated with DRB1*0401 allele and DRB1*03-DRB1*04 haplotype, while the dominant protection was connected with DRB1*0403 and nonDRB1*03-nonDR*04 haplotype. Our study suggests that the estimation of HLA DRB1*04 allele subtypes serves as a marker for better identification of subjects at risk of IDDM development in the first degree relatives of IDDM patients. PMID- 10680452 TI - [Clinical evaluation of amino acid solution]. AB - Glutamine, a conditionally essential amino acid, is important for immune function. It is now being formulated for incorporation in to total parenteral nutrition (TPN). The aims of this study were to examine the effect of glutamine administration on clinical status, body composition, protein synthesis and immune function in critically ill patients with sepsis. Eleven patients were included into the study. Seven of them have been on conventional TPN for 8-63 days without significant clinical and nutritional improvement. Glutamine supplemented TPN was implemented for 10 subsequent days. Before, during and after the study venous blood samples were taken for the cellular immunity examination, and for the measurement of plasma albumin, transferrin and triglycerides levels. Nitrogen balance was calculated every day during the study. No side effects were noted. Patients receiving amino acid solution revealed improved nitrogen balance, body composition and body water distribution. Plasma proteins concentrations and immunological indices significantly increased during ten days TPN with Glamin. We confirm the beneficial effects of amino acid solution-supplemented TPN on nitrogen balance, plasma proteins, and immune status of critically ill patients. PMID- 10680453 TI - [Use of plasmapheresis in patients with AIDS and peripheral polyneuropathy]. AB - The plasmapheresis is procedure removal of plasma and simultaneously, together with plasma, toxins, incorrect metabolites, antigens and immunological complexes. In patients with HIV infection often appearance periphery polyneuropathy. This is connected with autoantibody action on the damage of neurone. The aim of this study was evaluation effectiveness of plasmapheresis in treatment patients with AIDS and polyneuropathy. Seven patients with AIDS--C3 and polyneuropathy coexistence were treatment by plasmapheresis. The symptoms of polyneuropathy decrease in all patients. Were don't influence on immunological system and side effect performance plasmapheresis. PMID- 10680454 TI - [Levels of proinflammatory cytokines: IL-1, IL-6, IL-8, TNF-alpha and receptor IL 6R in Lyme borreliosis]. AB - We estimated serum concentrations of cytokines IL-1, IL-6, IL-8, TNF-alfa and IL 6R of patients with diagnosed Lyme disease treated with beta-lactam antibiotics. Detection of proinflammatory cytokines was performed in ELISA tests. The examination was performed before and after treatment. Comparison with control group stated statistically significant higher concentration of IL-1 and IL-6 before and after treatment. There were no differences in concentration of TNF alfa, IL-8 and IL-6R. Comparing concentrations of cytokines before and after treatment there was no differences either. Lack of changes in concentration of proinflammatory cytokines during beta-lactam therapy could be explained by too short period of therapy or immunologic background of inflammatory process in Lyme disease which was only initiated by spirochete Borrelia burgdorferi. PMID- 10680455 TI - [Squamous cell carcinoma antigen for monitoring patients with head neoplasm]. AB - The authors present results of evaluation of squamous cell carcinoma antigen (SCC) in 73 persons (35 patients with head carcinoma and 38 healthy persons as control group). We analysed a concentration of SCC in preliminary study and in monitoring, depending on clinical advances of tumour, mass of primary tumour and metastases to lymph nodes. We observed high sensitivity (SE) (63%), high specificity (SP) (97.4%) and predictive values: positive PV(+)--95.6%, negative PV(-)--74% of SCC in preliminary study. These values changed to SE--83.3%, SP- 92.3%, PV(+) 90.9%, PV(-)--85.7% in monitoring. Our results indicate that evaluation of SCC have high value in monitoring patients with head carcinoma. PMID- 10680456 TI - [Clinical and histopathologic picture of lichen sclerosus treated with clobetasol propionate]. AB - Effects of treating of the vulvar lichen scleroses by topical using of clobetasol propionate. The period of observation was 1 year. All patients showed clinical improvement or full regression depending on the period of disease. In all cases histopathologic examination was performed. PMID- 10680457 TI - [Monilethrix--rare syndrome of structural hair abnormalities]. AB - Monilethrix is a rare structural disorder of hair. Characteristic abnormalities in the form of alternating thinning and fusiform thickening are observed in most of hair shafts that we call beaded hair. Macroscopic estimation shows lustreless, dry, rough, fragile hair. Trichological examination usually reveals a considerable percentage of anagenic hair. According to our own experiences and literature data systemic therapy (vitamins) and topical treatment (desquamative ointments) are not effective sufficiently. Spontaneous regression of symptoms often appears with time. Five cases of familial occurrence of monilethrix have been presented. PMID- 10680458 TI - [A case of hamartoma in the larynx]. AB - The authors describe a rare case of hamartoma in the larynx of a 31 years old man. The egg sized tumor was situated in the left aryepiglottic fold filling the lower pharyngeal space. The unfortunate location, patient disdain as well as late diagnosis and proper management caused the histologically benign tumor to enlarge and block the laryngeal entrance that endangering life. After initial tracheotomy the tumor was successfully removed through the mouth. PMID- 10680459 TI - [The role of some cytokines in the etiopathogenesis of leukemia]. AB - Some cytokines play important role in etiology and pathogenesis of leukaemia. Most of them more stimulate than inhibit the proliferation of leukaemia cells. PMID- 10680460 TI - [An attempt to clarify the etiopathogenesis of chronic lymphatic B-cell leukemia]. AB - The paper presents current opinions about aetiology and pathogenesis of chronic lymphatic B-cell leukaemia. PMID- 10680461 TI - [A review of methods for organizing nutritional education for hospitalized patients]. AB - Diet-related diseases are diagnosed very frequently among inpatients and result in high mortality rate. Most commonly, the severity of illness or poor prognosis can be reduced or diminish by change of the lifestyle or simply-eating behavior. Across many countries there is conducted research of nutrition education potentials. It is usually applied to patients during their staying in hospital and after. A review of methods of nutrition education for hospitalised patients was done. The methods can be classified into three categories: instructional, motivational and behavioral. The effectiveness of the methods was the main concern. There were two major ways to evaluate this effectiveness: direct approach (laboratory or anthropometry tests) and indirect approach (questionnaires, food diary). An advantage of nutrition education providers' employment was discussed. Many studies show that nutrition education is an effective method in secondary prevention of diet-related diseases. It is promising to disseminate these kind of efforts in Poland. PMID- 10680462 TI - [Treatment with iodine radioisotopes of benign thyroid disease]. PMID- 10680463 TI - [Hematopoietic stem cells I. Basal aspects]. AB - The knowledge on basic aspects of hematopoietic stem cells (HSC) has markedly increased during the last decade to the extent that such cells are now routinely employed therapeutically in order to restore hematopoiesis following myeloablative chemotherapy and irradiation in patients with malignant disorders. Different methods can be applied to characterize HSC. Thus, by immunophenotyping and flow cytometry it is possible to delineate subpopulations of cells enriched for HSC. However, since phenotypic characteristics associated with HSC do not necessarily correlate to their developmental potential, measurements of HSC also rely heavily upon functional in vitro and in vivo assays, which demonstrate the presence of HSC through their ability to proliferate and differentiate in a clonogenic fashion. In this review we describe the assays developed to characterize and quantitate immature HSC, which form the basis for their clinical application. PMID- 10680464 TI - [Hematopoietic stem cells II. Diagnostic and therapeutic aspects]. AB - Characterization and isolation of haematopoietic stem cells (HSC) have resulted in their clinical application in patients with malignant disorders and--through gene therapeutic initiatives--also in the treatment of inherited diseases. Autologous stem cell transplantation (ASCT), which was introduced because of the high number of relapses in cancer patients in remission, involves dose intensification (conditioning), which induces myeloablation. In this setting, reinfusion of HSC is performed to restore haematopoiesis. Flow cytometric determination of CD34+ cells and clonogenic assays for committed myeloid HSC (CFU GM) are vehicles for quality control of the harvested HSC material and are integrated into the ASCT programs. Moreover, harvest of HSC and purification of CD34+ cells enables new treatment options such as removal of cancer cells from grafts, optimization of gene transduction as well as ex vivo expansion of HSC before reinfusion. In conclusion, the expanding insights into HSC in the 1990's have already been translated into valuable diagnostic and therapeutic modalities. PMID- 10680465 TI - [The connection between the intima-media-complex and atherosclerosis]. AB - In atherosclerosis several years will pass from development of the first fatty streaks to the first clinical event. In epidemiological investigations of atherosclerosis it is therefore desirable to have a valid method for detecting atherosclerosis in the earliest stages, which is harmless and can be performed repeatedly without risks in the healthy volunteer. Measuring the intima-media complex (IMT) using ultrasound constitutes such a method. Results from international research are reviewed and it is concluded that the method should be applied in Denmark in atherosclerosis research. PMID- 10680466 TI - [Azathioprine treatment of Crohn disease]. AB - Azathioprine/6MP (AZA/6MP) is effective in long-term treatment (> 3 months) of Crohn's disease and superior to other established medical treatments. The optimal dose remains to be defined. So far, effect has been demonstrated with 2-2.5 mg azathioprine/kg/day, but not with 1 mg/kg/day. A disease controlling effect has been demonstrated during up to four years of continuous treatment, after which data remains to be established. As part of remission-inducing combination therapy the effect of AZA/6MP can not be detected until two-three months after treatment start. High dose intravenous AZA/6MP administration does not shorten this interval. Reversible dose dependent side effects may require dose reduction or termination of treatment. Reversible dose independent side effects exclude further or repeated treatment. Some 10-15% stop treatment due to side effects. There is no increased death rate due to cancer in AZA/6MP treated Crohn patients. When giving the above full dose of AZA/6MP, monthly blood tests are recommended for the entire treatment period, more often during the first three months. PMID- 10680467 TI - [Radioiodine therapy of benign thyroid disease in Denmark]. AB - The aim of the study was to evaluate differences in the use of radioactive iodine in the treatment of benign thyroid disease in Denmark. A questionnaire was distributed to all departments in Denmark which administer radioiodine in the treatment of benign thyroid disease (n = 20). Radioiodine is used for patients with toxic nodular goitre and for patients with relapse of toxic diffuse goitre. Four departments did not use radioiodine for volume reduction in non-toxic goitre. Patient informations included very different recommendations regarding cautions in relation to radioiodine treatment. Radioiodine is widely used in the treatment of benign thyroid disease. We recommend a national standardization of the cautions in relation to radioiodine treatment. PMID- 10680468 TI - [Nitric oxide inhalation therapy in patients with acute respiratory distress syndrome. A retrospective analysis]. AB - Acute lung injury in the form of the acute respiratory distress syndrome (ARDS) continues to carry a high mortality. Recently, inhaled nitric oxide (NO) has been used in ARDS patients with severe hypoxaemia. We present a retrospective analysis concerning 33 ARDS patients treated with inhaled NO during a period of four years from 1994 to 1998. Patients were young, mean 43 years, had poor oxygenation (PaO2/FiO2 index 67 mmHG) and a lung injury score above 2.5. Eighty-two percent responded immediately to NO, 9% at a later challenge and 9% were non-responders. Mortality for the whole group was 67% and to be expected considering the severity of disease. For the group of patients with ARDS as a complication of pneumonia mortality was only 40%. In one patient therapy had to be stopped due to formation of NO2 even at very low concentrations of inhaled NO. We conclude that inhaled NO is effective in improving oxygenation in patients with ARDS. The study does not allow us to conclude anything about mortality, which was probably unchanged by this novel therapy. PMID- 10680469 TI - [Nitrogen oxide inhalation in critically ill patients]. AB - Inhaled nitric oxide (iNO) has been extensively used in the treatment of severe hypoxaemic respiratory failure and/or pulmonary hypertension. Although the majority of studies have almost consistently demonstrated a beneficial effect on oxygenation and a reduction in pulmonary artery pressures, none of the randomised trials have reported a reduced mortality using iNO on patients with severe respiratory failure. In neonates with severe respiratory failure and pulmonary hypertension, however, iNO can reduce the need of extracorporal membrane oxygenation. Prior to using iNO on adult patients we suggest other measures to be taken (i.e. optimising ventilator settings, ventilation in prone position). PMID- 10680470 TI - [Prognosis after primary acute myocardial infarction]. AB - Results from many clinical trials have changed treatment strategies, but due to considerable selection caused by extensive inclusion- and exclusion criteria the results are only valid for smaller groups, while the majority of the patients still suffer from high mortality with conservative treatment. We have during eight years followed 933 patients with first myocardial infarction and found that the group who tolerated treatment with thrombolytic agents had a lower mortality than the rest. On the other hand, infarct patients are generally old, often with concomitant chronic diseases which affect mortality, and this in combination with heart-failure and cardiac arrhythmias determines outcome. Better prophylactic and treatment measures are needed to improve survival of ischaemic heart disease. PMID- 10680471 TI - [Staphylococcus aureus endocarditis in Denmark 1982-1991. Clinical picture, complications and antibiotic therapy in non-drug addicts]. AB - The central S. aureus surveillance in Denmark made it possible to analyze the clinical features of S. aureus endocarditis in a nation-wide population of non drug addicts. Almost all cases of bacteraemia with S. aureus are reported to the Staphylococcus laboratory, Copenhagen. The medical records were reviewed in cases from 1982 to 1991 in which the diagnosis of endocarditis was reported or suspected. Two hundred and sixty patients fulfilled the diagnostic criteria. In 83 patients the diagnosis of endocarditis was not suspected clinically. The overall mortality rate among those patients whose disease was diagnosed clinically was 46% and significantly associated with late congestive heart failure, age and involvement of the central nervous system. A more frequent use of echocardiography as a screening method seems essential to improve the prognosis of patients with S. aureus endocarditis. Involvement of the CNS constitutes a relative indication for early valve replacement. PMID- 10680472 TI - [Diet, cancer and health--a population study and establishment of a biological bank in Denmark]. AB - In order to test hypotheses on diet and the risk of cancer, a prospective cohort study was established. A total of 57,055 persons living in Copenhagen and Aarhus, between 50 and 65 years of age, visited a study clinic between December 1993 and May 1997. The participants provided questionnaire data on diet and lifestyle. Furthermore, anthropometric measurements, blood pressure and biological material were collected. All participants will be followed by linkage to health registries including the Cancer Registry and by self-administered follow-up questionnaires. The purpose of this publication is to describe the data-base, which will be available for research in the years to come including the results of the first two years of follow-up. PMID- 10680473 TI - [Perioperative administration of recombinant human erythropoietin in colorectal cancer surgery. A prospective, randomized, double-blind placebo controlled study]. AB - One hundred patients scheduled for elective colo-rectal cancer surgery, and with a preoperative haemoglobin level < or = 8.5 mmol/l were included. Eighty-one patients could be evaluated. Thirty-eight patients received r-HuEPO in a dose of 300 IU/kg body weight on day four before surgery and 150 IU/kg, daily, for the following seven days, and 43 patients received placebo. In addition, all patients received daily doses of 200 mg iron, orally, for four days before surgery. On the day of surgery and until discharge the haemoglobin concentration was significantly higher in the erythropoietin group compared to the placebo group. The number of blood transfusions given was significantly lower in the erythropoietin group with a mean of 0.3 units per patient (0-6) compared to 1.6 units (0-9) in the control group (p < 0.05). The clinical implications of these findings has yet to be assessed. PMID- 10680474 TI - [Mycobacterium avium complex infection in a patient with idiopathic CD4+ T lymphocytopenia]. AB - Primary cutaneous infections with MAC are extremely rare. We report a case of primary cutaneous infection with MAC, in a 69 year-old HIV-negative male. Idiopathic CD4+ T-lymphocytopenia was diagnosed. PMID- 10680475 TI - [Rupture of an isolated iliac aneurysm]. AB - Isolated iliac artery aneurysm threatening to rupture is a rare condition with a poor prognosis if not dealt with by the physician in a brisk and effective manner. Symptoms from an iliac aneurysm can masquerade as a range of symptoms from neighbouring organs or be almost asymptomatic before rupturing. A case of a 81 year old man, with symptoms of dizziness, fatigue and light lower abdominal pain, who developed sudden cardiovascular collapse, is presented. PMID- 10680476 TI - [Bilateral rupture of the quadriceps tendon in a healthy individual]. AB - A case of bilateral rupture of the quadriceps tendons in a 59 year-old man without any known systemic disease is presented. The ruptures occurred while he was descending a staircase, after which he fell. Repetitive microtrauma was suspected as the etiological reason for rupture. PMID- 10680477 TI - [Picture of the month. Hepatopulmonary syndrome]. PMID- 10680478 TI - [Leptin in arterial hypertension]. PMID- 10680479 TI - [From scientific recognition to practical clinical guidelines]. PMID- 10680480 TI - [Innate mechanisms during pregnancy]. PMID- 10680481 TI - [Demand and offer of coronary angiography in Denmark]. PMID- 10680482 TI - [Polyneuropathy in critically ill patients]. PMID- 10680483 TI - [A method of unilateral operation for early cleft palate repair]. AB - OBJECTIVE: This paper presents a new method for cleft palate repair. METHOD: Six changes have been made to the conventional procedures. After all of these six changes have been carried out on one side of the palate, the operated side becomes completely relaxed. It can be moved both posteriorly and medially to lengthen the side of the palate and to contact with the cleft margin of the non operated side without tension. Obviously, there is no need to perform the same surgical maneuvers on the other side again. The cleft is then closed by layers. RESULT: Totally 151 cases of unilateral and bilateral cleft palate have been repaired with this method. The age of these patients ranged from 4 months to 5 years. Postoperatively, there was neither one death nor one dehiscence. All of the baby patients who received treatment before they began to speak had good quality of speech or near normal speech function. CONCLUSION: In the unilateral operation, the surgical trauma, blood loss and time required for the operation are all less than that of conventional procedures which operate on both sides. It is a safer method for early cleft palate repair. PMID- 10680484 TI - [Cleft palate repair with bilateral buccal musculomucosal flaps]. AB - OBJECTIVE: A method for cleft palate repair was used for the purpose of diminishing surgical interference with maxillary growth. METHOD: Sixteen cases of cleft palate were repaired with bilateral buccal musculomucosal flaps. RESULT: The operation results in all cases were satisfactory. CONCLUSION: It is concluded: 1. The method does not traumatize the periosteum of the hard palate, thus alleviating postoperative maxillary deformity; 2. The buccal musculomucosal flap is generally long enough to reach across the incisive foramen and maintain good blood circulation; 3. The method benefits reconstruction of the levator sling and facilitates velopharyngeal closure. PMID- 10680485 TI - [Correction of nasal septal deformities of the unilateral cleft lip]. AB - OBJECTIVE: Septal deformities are very common in patients with unilateral cleft lip nose. The deformities are influential on the morphology of the external nose and respiratory function. The study was conducted to correct the septal deformities. METHOD: The operation procedure includes correction and replacement of the deviated septum to the middle position; correction of the displaced alar cartilage with the suspension method and correction of soft tissue deformities of the nose and lip. Twelve patients were treated with this method from 1994 to 1995. RESULT: Postoperative follow-up of 3-12 months demonstrated satisfactory results. CONCLUSION: Correction of septal deformities plays a very important role in the operation for the unilateral cleft lip nose. PMID- 10680486 TI - [Anatomic study of the musculus depressor septi nasi in unilateral cleft lip nasal deformity]. AB - OBJECTIVE: This study was to investigate the nasal musculature and its relationship with the deformities of the unilateral cleft lip nose. METHOD: The anatomic and histological examination and comparison of the musculus depressor septi nasi (MDSN) were performed in 33 normal cadavers and 30 patients with unilateral cleft lip undergoing the operation for secondary nasal deformities. RESULT: It was found that the MDSN in the cleft side of a unilateral cleft lip nose is lacking. CONCLUSION: Because of the anatomic deficiency on the cleft side, the function of the normal side is pathologically increased. The functional imbalance between the normal and abnormal side may bring about dislocation and malformation of the base of nasal columella as well as the septal structures. PMID- 10680487 TI - [The effect of tangential excision on the local IL-8 release and wound inflammatory response in deep-partial thickness burn]. AB - OBJECTIVE: The aim of this study is to investigate the effect of tangential excision on the local IL-8 release and wound inflammatory response. METHODS: The tissue specimens, which were taken from 7 patients with deep partial thickness burn before and after operation, were both cultured and histopathologically examined. The level of IL-8 in the culture medium was assayed, meanwhile local inflammatory response of tissue samples was evaluated. RESULTS: The result showed that the level of IL-8 is much lower in the postoperation wounds, while in the non-operation wounds the neutrophil infiltration was marked, with enlargement in necrotic area and disappearance of skin appendages. CONCLUSION: The findings suggested that tangential excision could reduce the local production of IL-8 and decrease local inflammatory response, which is helpful to prevent the progressive necrosis of burn wound. PMID- 10680489 TI - [Mechanisms of early gastro-intestinal ischemia after burn: hemodynamic and hemorrheologic features]. AB - OBJECTIVE: To study the mechanisms of early gastro-intestinal ischemia developed in acute burn period and its relationships with hemodynamic and hemorrheologic changes. METHODS: Twelve pigs were randomly allocated into two groups: group C, a sham group that was subjected to all surgical procedures except burn; group B, 30% TBSA cutaneous thermal injury, and was resuscitated with Parkland formula one hour after burn. RESULTS: MAP remained stable after burn, but RAP, MPAP, PAWP and CI decreased significantly to the lowest level 4-8 hours after burn, and recovered after resuscitation 24 hours postburn. Intramucosal pH declined immediately (1 hour after burn) and remained abnormal throughout observation period. Portal venous blood flow demonstrated similar changes as pHi, and correlated well with intestinal pHi. Whole blood viscosity and plasma viscosity in portal venous blood elevated obviously after burn. CONCLUSIONS: 1. GI ischemia occurred early and recovered slowly during burn shock phase with conventional resuscitation regime. 2. GI ischemia correlated significantly with portal venous blood flow, but did not with systemic hemodynamic variables. 3. Hemorheologic changes in portal venous blood may exaggerate ischemia injuries. PMID- 10680488 TI - [The influence of adhesion molecules expression to neutrophil aggregation in liver after severe burns]. AB - OBJECTIVE: To elucidate the mechanism of neutrophil aggregation in the liver and the influence of the expression of adhesion molecules after severe burns during early postburn phase, and to explore simultaneously the mechanism of liver damage after severe burns. METHODS: 1. The morphology of the liver and the activation of MPO were observed. 2. The human hepatocytes and hepatic sinusoidal endothelial cells (HSECs) were isolated and cultured, the flow cytometry and micropipette aspiration system were used to observe dynamically the change in intercellular adhesive force between neutrophils and hepatocytes under the influence of burn patient sera. Also the changes in the expression of P-selectin and ICAM-1 on the hepatocytes and HSECs under the influence of burn patient sera were studied. RESULTS: 1. MPO activation was significantly higher than that in the control group, and reached its peak at 6 hours. Three pieces of liver tissue at 6, 12, 24 hours were examined, and it was found that the Disse's spaces were wide and filled with a great number of PMNs. 2. A constitutive expression of P-selectin and ICAM-1 was found in normal hepatic sinusoidal endothelial cells but not in normal hepatocytes. Under the influence of burn patient sera, the expression of P selectin increased significantly at 1 hour (P < 0.01) in hepatic sinusoidal endothelial cells but not in hepatocytes, and the expression of ICAM-1 increased significantly at 1 and 2 hour (P < 0.01) in hepatocytes and hepatic sinusoidal endothelial cells. 3. In the presence of burn patient sera, the intercellular adhesive force between neutrophils and hepatocytes reached its peak level at 2 hour after co- cultivation and declined thereafter due to the aggravation of hepatocytic injury. After using ICAM-1 Mab in cultured hepatocytes, the adhesive force reduced significantly. CONCLUSION: Neutrophil aggregation in the liver was confirmed after severe burns during early postburn phase. The expression of P selectin and ICAM-1 on the hepatocytes and HSECs played key roles to neutrophil aggregation in liver after severe burns during early postburn phase, inducing the participation of neutrophils in hepatocyte injury. PMID- 10680490 TI - [Relationship between plasma D(-)-lactate levels and acute intestinal injury in rats following ischemia-reperfusion]. AB - OBJECTIVE: To determine the kinetics of plasma D(-)-lactate levels in both portal and systemic circulations, and to examine whether elevated plasma D(-)-lactate would correlate to intestinal injury in rats subjected to acute intestinal ischemia-reperfusion. METHODS: Anesthetized rats underwent 75 minutes of superior mesenteric artery occlusion followed by 6 hours of reperfusion. Plasma D(-) lactate levels were measured by an enzymatic spectrophotometric assay. RESULTS: It showed that intestinal ischemia for 75 minutes resulted in a significant elevation in D(-)-lactate levels in portal vein blood compared to baseline values (P < 0.05). Plasma D(-)-lactate levels had a tendency to further increase after reperfusion up to 6 hours. Similar alterations in D(-)-lactate were also found in systemic circulation, there were no significant differences between the portal and systemic circulation at any time point. Moreover, the histopathological evaluation scores were significantly correlated to the portal D(-)-lactate levels in animals at various time points (r = 0.415, P < 0.01). In addition, a remarkable rise of endotoxin concentration within portal vein was already found at the end of 75-minute ischemia (P < 0.05), reaching a peak at 2 hours post reperfusion. CONCLUSION: These data suggest that acute intestinal ischemia is associated with failure of mucosal barrier resulting in increased plasma D(-) lactate levels in both portal and systemic blood. The subsequent reperfusion might cause further increase in D(-)-lactate levels, which correlated to the histopathological alterations. Plasma D(-)-lactate may be a useful marker of intestinal injury following both ischemia and reperfusion insults. PMID- 10680491 TI - [A new burn wound covering-fibroin membrane]. AB - OBJECTIVE: This study was aimed at determining the physical and biological properties of fibroin membrane and evaluating its clinical effects. METHODS: 1. The physical and biological properties of fibroin membrane and partial thickness porcine skin were determined with various instruments. The determinations included water ratio, tensile strength, flexibility, water permeability, adhesive force and so on. 2. CLINICAL OBSERVATION: Two wounds with same depth in the same patient were selected. One was covered by fibroin membrane. The other one was treated with 1% SD-Ag cream. The changes in these wounds were observed and compared. RESULTS: This membrane had nearly same physical and biological properties compared with partial thickness porcine skin. It had no toxicity, irritation, and antigenicity. It possessed certain amount of water permeability and could adhere to the wound. Because of its good transparency, any change in underlying wound could be directly observed. It was helpful not only in local therapy but also in study of wound healing. CLINICAL OBSERVATIONs showed that this membrane had good protecting effect to wound. Its application could obviously relieve pain of wound and the healing time was 3-5 days ahead than that of the control wound. CONCLUSION: This membrane was an ideal wound covering and was worth being popularized. PMID- 10680492 TI - [The influence of different concentrations of hydrofluoric acid on blood calcium level in rats]. AB - OBJECTIVE: To evaluate the effects of different concentration of hydrofluoric acid (HF) on local skin and calcium level of serum. METHODS: SD rats were divided into two groups which were wounded by 20% and 40% percent of HF respectively. Samples of blood and wound tissue were harvested at different postburn time for the analysis of the calcium level of the serum and histological study. RESULTS: It was found that twenty percent of HF was enough to cause a skin damage, and might bring about fatal hypocalcemia after a prolonged contact. High concentration of HF (40%) could cause deep tissue necrosis within a short time, and result in a fatal hypocalcemia within 24 hour even in the case of a small area injury. CONCLUSION: It is important to treat the patient with HF injury as early as possible. Sufficient calcium must be applied guided by laboratory study in order to prevent the fatal hypocalcemia. PMID- 10680493 TI - [The effect of growth hormone on wound healing rate in adult burns]. AB - OBJECTIVE: To determine the wound healing effect of rHGH in adult burn patients. METHODS: 16 patients with burn wounds covering over 60% of total body surface (TBSA) were enrolled in this placebo controlled prospective study. They were comparable in nutrient intake, TBSA, and full thickness burn area. rHGH group patients were given rHGH subcutaneously in the dose of 0.3u.Kg-1 at 8 am each morning for 10 days beginning from POD 1. The control group patients were given normal saline as placebo. All the patients received scar excision within 4 days postburn, and the excision wounds were covered with autologous skin pulp grafting. Serum amino acid profile was analyzed at day 1 and day 20 post burn. Healing time of burn wound area and donor site was recorded. Wound healing rate was assessed at day 30 after day. RESULTS: 1. The healing time of autologous skin pulp grafting and donor site, and the length of hospital stay were significantly shorter in GH group patients than control group. 2. The amino acid profile showed no difference between two groups at day 1 and was significantly better in GH group at day 20. CONCLUSION: rHGH could enhance the wound healing rate, improve amino acid profile, and reduce the length of hospital stay of severe burn patients. PMID- 10680494 TI - [Experimental study on denervated skeletal muscle autotransplantation in the rhesus monkey]. AB - OBJECTIVE: This study was to provide a scientific basis for clinical application of denervated muscle transplantation. METHODS: A rhesus monkey was used in this experiment. The denervated extensor digitorum brevis (EDB) and the extensor hallucis brevis (EHB) of the rhesus monkey were autotransplanted in the sacrospinal muscle, and they were biopsied at intervals and stained with HE, Bielshowsky and enzyme. The specimen were examined with light and electro microscope. RESULTS: The metabolism of the denervated muscle was lower than the normal. There was muscle regeneration and growing nerve fibers in the graft 12 weeks after transplantation. CONCLUSION: Denervated muscle can regenerate in oxygen-deficient environment. If the denervated muscle is in contact with normal muscle, the nerve of the recipient muscle will grow into the graft. PMID- 10680495 TI - [A study on pearl fat transplantation--concentration of basic fibroblast growth factor (bFGF) and carrier of sustained delivery selected by orthogonal design]. AB - OBJECTIVE: The experiment was designed to solve the problem that free fat transplantation survived at low percentage. METHODS: Based on physiochemical and biological characteristics of bFGF, fibrinogen was used as its carrier of sustained delivery. We applied bFGF and fibrinogen in pearl adipose transplantation. Different concentrations of bFGF and fibrinogen were used. The relation between the concentration of bFGF and fibrinogen and their effects on fat graft maintenance were observed. RESULTS: At three months after transplantation, the weight of the fat graft varied from 98% to 225%. The latter was approximately consistent with synchronous increase of the body weight of the animal. Weight increase of the graft was closely related to the concentration of bFGF and the sustained delivery carrier. When bFGF in the concentration of 4000 U/10 microliters and fibrinogen of 1000 mg% were applied, the fat graft obtained the greatest weight increase. Histologically, the structure of the transplanted fat was normal with fibrotic envelope and mature adipocytes. CONCLUSION: bFGF with fibrinogen as its carrier of sustained delivery favors the survival of fat graft and weight increase. The effects of bFGF and fibrinogen depend on their concentration. PMID- 10680496 TI - [An experimental study on promoting reconstruction of microcirculation of the ischemic skin flap with basic fibroblast growth factor]. AB - OBJECTIVE: This study was to determine whether the basic fibroblast growth factor (bFGF) could promote reconstruction of microcirculation in the ischemic portion of a random skin flap. METHODS: Five methods were used in the experiment: measurement of the survival area of the skin flap; light and electron microscopic studies; immunohistochemical observation of the endothelial cell and microvessel image analysis technology (IAT). RESULT: The survival area and microvessels in the skin flap were significantly increased in the treated group. The results indicate that bFGF can stimulate the proliferation and differentiation of endothelial cells in vivo and promote the microcirculation of the ischemic flap. CONCLUSION: bFGF could promote reconstruction of microcirculation of the ischemic skin flap. PMID- 10680497 TI - [Application of the free parascapular flap in children]. AB - OBJECTIVE: To improve surgical results we have applied the free parascapular flap in children for facial or cervical burn scar contracture. METHOD: Eight children aged from 6 to 9 years received free flap transfer. Based on the defect and the distribution of the cutaneous branches of the circumflex scapular vessels, the flap was designed with its size ranging from 15 cm x 8 cm to 22 cm x 6.5 cm. RESULT: Free flap transferring with microvascular anastomosis was successful in all cases. The average operation time was 5 hours with average blood loss being about 100 ml. CONCLUSION: Free flap transplantation is safe and beneficial in children when the technique and skill of microsurgery and plastic and reconstructive surgery are mastered and intraoperative blood loss is controlled under 1% of the total blood volume. The vertically oriented parascapular flap can not only provide proper tissue for reconstraction but also minimise the donor site morbidity. PMID- 10680498 TI - [Surgical treatment of excessive hyperplasia symptom of skin keratosis at the limb ends]. AB - OBJECTIVE: We introduce the experience of treating 10 cases with excessive hyperplasia of skin keratosis at the limb end. There were 9 cases of palmar and plantar keratosis and 1 case of epidermodysplasia verruciformis. METHOD: All cases received the operation of lesion excision and skin grafting except 2 cases who received skin flap for defect covering after lesion excision because of deep tissue exposure and postradiation ulcer. RESULT: The operative effects were satisfactory after 8 years of follow-up. No recurrence was found in all cases and all patients resumed normal activity. CONCLUSION: The effect of the skin flap is better than skin graft at the plantar area. Because of lack of fibrous septum in the subcutaneous tissue, the flap, often moving during walking, is liable to trauma and ulceration. The operation area must be well protected. PMID- 10680499 TI - [A study of photographic standardization in rhinoplasty]. AB - OBJECTIVE: A method of photographic standardization for rhinoplasty is presented. METHOD: To delineate the nose objectively before and after rhinoplasty, we designed a method to standardize picturing by using the points and lines in the face and camera in nasal photography. RESULT: With the help of anatomic points in the face and lines in the camera, we can maintain the camera at a constant position to the face. In this way, the nasal picture is more comparable and instructive. CONCLUSION: This method has been proved to be helpful in rhinoplasty. PMID- 10680501 TI - [Application and characteristics of biomaterials commonly used in plastic surgery -current status and prospects]. PMID- 10680500 TI - [Progress in the study of wound dressing]. PMID- 10680502 TI - [Scopes and applications of LeFort III advancement osteotomy]. AB - In order to correct craniofacial retrusion and malocculusion caused by Crouzon syndrome, we employed LeFort III advancement osteotomies in 21 cases in last 10 years. The differences between craniofacial appearances and cephalometric analyses pre- and postoperatively showed that good results were achieved in all cases. No severe complications in these series were found. PMID- 10680503 TI - [The primarily study on the correlation between the cephalometric characteristics and the results of ploysomnography]. AB - A study on correlation between temporomandibular joint ankylosis and Obstructive Sleep Apnea Syndrome (OSAS) was conducted. The statistic results demonstrate that there is significant correlation between (TA + SPA)/OPA and AI, and there is negative correlation between PAS and AI, (TA + SPA)/OPA and SaO2min by correlation and step regression between the cephalometric analysis and polysomnogram of fourteen cases of OSAS patients. PMID- 10680504 TI - [A positioning device for oral paralleling technique]. AB - OBJECTIVE: To design a reliable method for standardized paralelling dental projection, for comparison of the tiny variations of alveolar bone during the treatment of the periodontal disease. METHODS AND MATERIAL: Periapical radiographs in 29 pairs of films with this device and 14 pairs of films with traditional method on dried skulls were comparatively analysed by digital subtraction. Standard deviations and angle variations of subtracted image were studied comparatively between this method and traditional paralleling technique. RESULTS: Significant difference of reproducibility between two groups was showed. The reproducibility of the successive pictures was apparently increased when projected with positioning device. PMID- 10680505 TI - [Quantitative studies of oncogene ras P21 and P53 gene protein expression in the benign and malignant pleomorphic adenomas of salivary gland]. AB - The oncogene ras P21 and P53 gene protein expression in the benign and malignant pleomorphic adenomas of salivary gland were quantitatively studied by flow cytometry and cellular immunofluorescence staining technique. The results showed that the ras P21 expression was found in 78% and 100% of the benign and malignant pleomorphic adenoma, respectively. The P53 protein was detected in 81% and 100% of the benign and malignant pleomorphic adenoma, respectively. The ras P21 and P53 protein were negative in the normal parotid tissue. The obvious different statistical significances of the fluorescence indexes (FI) in ras P21 and P53 expression were found between the benign and malignant pleomorphic adenoma (P < 0.001, respectively) as well as between the normal parotid tissue and the benign pleomorphic adenoma (P < 0.001, respectively). These results indicate that activation of ras oncogene and the mutation of P53 gene may play important roles in the tumourigenesis and the malignant transformation of the pleomorphic adenoma. PMID- 10680506 TI - [The effect of artificial traumatic occlusion on the pulp and periodontium in rat molars]. AB - In order to study relationship between the period of occlusal trauma and pulpal and periodontal tissues, 44 rats were used in experimental occlusal trauma. The pulpal and periodontal tissues of rat molars were observed by histological examination and micrographic examination. The results showed that the damage of pulp became more serious as time went on, but periodontium appeared adaptive changes. The same changing area in periodontal tissues were observed between histological section and micrographic film. The results provide experimental evidences for the pathology and pathogeny of pulpitis and perioapical periodontitis caused by occlusal trauma. PMID- 10680508 TI - [Anatomy of the cartilages of the nose with secondary deformity associated with unilateral cleft lip]. AB - To explore the influence of displacement and hypoplasia of nasal cartilages to cleft lip nose, 38 patients with secondary cleft lip nasal deformity were randomly chosen. Measurement of length and width of the nasal cartilages was made, and places of them were inspected. The displacement of the cartilages was found in each nose, and hypoplasia of cartilages was in those noses whose deformities were more severe. The conclusion is that both displacement and hypoplasia of nasal cartilages are causing factors of nasal deformity. PMID- 10680509 TI - [Subperiosteal implantation of block coral on the rabbit cavarial bone]. AB - The aim of this study was to evaluate osteoconductive effect of coral implanted on surface of the cortical bone. The 2 mm thick block corals were implanted on the surface of one side of rabbits cavarial bone, the another side were used as blank control. The specimen was examined by light microscopy, histometry and scanning electron microscopy at 2, 4, 8, 12 weeks after operation respectively. The results show that the new bone growth into the pore of the coral at 2 weeks after implantation. The coral was progressively absorbed and replaced by new bone. Up to 12 weeks, most of the coral was absorbed and newly formed bone had been remodeled. The reconstructed height of new bone in coral group was significantly higher than that of blank control and lower than the thickness of block coral. The study demonstrated good osteoconductive property of coral implanted on surface of cortical bone. PMID- 10680507 TI - [CT examination and diagnosis of oral and maxillofacial tumors invading the pterygopalatine space]. AB - CT findings in 33 cases (proven histopathologically) suffered from oral and maxillofacial tumors affecting the pterygopalatine space were retrospectively analyzed. The authors conclude that the main CT manifestations of this space involvement by tumors can be depicted as a soft tissue mass occupancy and the wall structures destruction. Tumors occurring in different locations of oral and maxillofacial areas have different features of CT appearances. In addition, the primary tumor of this space is probed from histogenesis and CT findings, and its diagnostic criteria are discussed. As a modality of imaging, CT has an important role for assessing the lesions of this fossa. PMID- 10680510 TI - [A preliminary report of histological study on TMJ reconstruction with autogenous costochondral graft]. AB - To study histologically the biological behaviour of the grafts and the reactions in the receipt area after reconstruction of temporomandibular joint (TMJ), we have reconstructed experimentally removed TMJs on purebred juvenile pigs by transplanting autogenous costo-chondral grafts. The findings of continuously survey of histologic changes in the receipt areas and the grafts showed that autogenous grafts healed well with the surrounding soft tissues, and induced the mesenchymal-like cells from the receipt region, rather than the periosteal cells which might be left, to differentiate and ossify both within and at the surface of them, and they themselves were gradually replaced by the new-formed bone; and that active osteogenesis could also be seen within the thick tissues far from the graft, which could not be the result of graft's induction. It may be concluded that the new bone, both from the induction of the graft which acts as a framework and somehow from the outside tissues, reconstructs the new condyle and determines the postsurgical growth of the mandible. PMID- 10680511 TI - [A quantitative study of microvascular density and proliferating cell ratio in buccal mucosa squamous cell carcinoma]. AB - By the ABC enzyme labelling method we studied the microvascular density and proliferating cell ratio in buccal mucosa squamous cell carcinoma (BMSCC) and in normal buccal tissues quantitatively. These two parameters in BMSCC are 208% and 335% (P < 0.05) respectively which are distinctly higher than those of the normal buccal tissues, indicating malignant features of the carcinoma. The distribution of proliferating cells, corresponds to the area where the microvascular density is high, mainly in the periphery and basal parts of cancer nests. There is a dependent relationship between them, showing a linear positive correlation statistically. The results confirm that angiogenesis and cancer cell proliferation promote each other. They are two important indexes of the local biological behavior of BMSCC and are very important to predict the malignant potential and to evaluate therapeutic effect for BMSCC. PMID- 10680512 TI - [Characteristics of lip-mouth region in smiling position from 80 adults with acceptable faces and individual normal occlusions]. AB - The Characteristics of Lip-mouth region including the soft and hard tissues in smiling position with frontal fixed position photographic computered analysis was studied. The examples were 80 adults (40 males and 40 females) with acceptable faces and individual normal occlusions. The age was between 17-25 years of age. The method adopting maximum smiling position to study the lip-mouth region is scientific, and the maximum smile line could be divided into three categories: low smile line-a-counted for 16.25% of the total samples; average smile line- 68.75%; high smile line--15%. PMID- 10680513 TI - [Immunohistochemical and quantitative analysis of expression of basement membrane in squamous cell carcinoma of the oral cavity]. AB - Fifty seven cases of oral squamous cell carcinoma (OSCC) were studied by immunohistochemical ABC method using type IV collagen and laminin antibodies and computer image analysis system to investigate the expression of basement membrane and its relation to clinicopathologic characteristics of the OSCC. The results demonstrated that: 1. in OSCC there were defects basement membrane in continuity of varying extents to complete loss of; 2. there was no correlation between the loss of basement membrane and tumor clinical stage (P > 0.05) by computer image analysis, while there was a highly significant correlation between the loss of basement membrane and histologic grading (P > 0.05). The well-differentiated tumor was associated with a low frequency of basement membrane loss. Therefore, the expression of basement membrane may be a useful parameter to evaluate tumor histologic differentiation. PMID- 10680514 TI - [Histopathological study on traumatic facial nerve injury in the rabbits]. AB - Six patterns of injuries (exposure, compression, crush, stretch, division and post-division anastomosis) were analysed in 136 facial nerve buccal branches of 68 rabbits in order to establish the experimental model for study of traumatic facial nerve injury. Changes of histopathology at definite time were studied. The results showed that: exposure can create degree I injury; compression, stretch and crush can create degree II-III injury; the prognosis in anastomosis group was obviously better than unanastomosis group. In moderate to serious injuries, changes of histopathology and ENoG were synchronized, but were not so in mild and later stage of serious injury. The 15th day postinjury was the best time of estimating the facial nerve function clinically. The area of injury would obviously influence the mylin lesion, while the quality and degree of injury would obviously influence the axonal lesion. PMID- 10680515 TI - [Expression of transforming growth factor beta 1 in keratinocytes of oral submucous fibrosis tissue]. AB - To explore significance of transforming growth factor beta 1 (TGF beta 1) in the pathogenesis of oral submucous fibrosis (OSF), TGF beta 1 mRNAs in keratinocytes of the paraffin embeded tissues of 25 OSF cases, 5 normals (NOR) and 10 oral lichen planus (OLP) were determined by the in situ hybridization technique. The result showed that there was an expression of TGF beta 1 mRNA in keratinocytes of 15 OSFs (60%). The positive expression of TGF beta 1 mRNA was mainly in kerationcytes of early and middle stage OSF. There was no expression in that of 5 NORs and 10 OLPs. The study suggests that keratinocytes of OSF tissue may synthesize and release TGF beta 1 which may play an important role in the pathogenesis of OSF and participate as a mediator in the pathogenetic process of OSF. PMID- 10680516 TI - [A three-year clinical evaluation of five light-cured composite resins in fillings of posterior teeth]. AB - In order to study the clinical performance and the evaluation method of light cured composite resin in filling for posterior teeth, two evaluation methods were used to evaluate 5 light-cured posterior composite resin fillings in 169 adult posterior class I cavity. Results showed that each evaluation method has its own advantages. The curative effect was declined with time in this study. Failures and defects were mainly occurred after 3 years. Secondary caries, loss of fillings and marginal stainings were the main reasons of failures. Compared with the effect of 1 year, the success rate after 3 years declined significantly, and further long-term clinical observation is needed. PMID- 10680517 TI - [A study on the cytotoxicity of six filling materials in vitro]. AB - This paper deals with the cytotoxicologic analyses on 6 filling materials with morphology of cells, ultraviolet light spectrophotometry and incorporation test using mouse L-929 fibroblasts labelled 3H-TdR. The results showed that the cytotoxicity of Silver amalgam and the Gallium-Silver alloys, which were produced by mixing the conventional dental alloys powder or high copper alloys powder with Gallium, was significantly stronger than that of light curing composites and the Gallium-Silver alloys that were produced by the spherical amalgam alloys powder and Gallium. It suggested that the level of mercury and copper in the alloys can influence their cytotoxic properties. PMID- 10680518 TI - [Perioral muscle forces and malocclusion]. PMID- 10680519 TI - [Power measurement of denervated muscle isografts with neurorrhaphy and nerve implantation in rats]. AB - The hypothesis was tested in this experiment that after 7 months of predencervation, the reinnervation of muscle grafts with neurorrhaphy results in greater recovery of force and power than with nerve-implantation. In a highly inbred strain of rats, soleus muscles were isografted. The donor muscles were either immediately denervated at the time of isografting or denervated 2, 4, 7 months prior to isografting. Soleus muscles from each donor group were transplanted into the right legs of hosts with either epineurial anastomosis (NR group) or nerve implantation (NI group). The contralateral soleus muscles of hosts served as controls. Sixty days after transfer, both right and left soleus grafts/muscles were evaluated for force and power measured in situ. The absolute force values were significant higher in NR group (61% of normal) than in NI group (40% of normal) in 2-month group but the result invenrsted in 7-month group, less than 20% and more than 20% of normal in NR and NI groups respectively. The reduced ability of grafts to generate force and power resulted from the different ways of reinnervation in denervated muscles and the period of predenervation. Maybe the nerve-implantation is better than the neurorrhaphy for reinnervating a long-term denervated muscle. PMID- 10680520 TI - [Interleukin-8, a regulator of inflammation in gingival crevicular fluid]. AB - The role of interleuking-8 (IL-8), a neutrophil-attracting and-activating cytokine, was investigated in gingival crevicular fluid (GCF). ELISA was used to detect the levels of IL-8 in GCF collected from 54 adult periodontitis (AP) patients (105 teeth) and 24 healthy subjects (54 teeth). The results showed that 1. The role of IL-8 was dependent upon the concentration of IL-8 in GCF. IL-8, in the low concentration (< or = 30 micrograms/L), was positively correlated with bleeding index (r = 0.36, P < 0.01). While in the high concentration (> 30 micrograms/L), was negatively correlated with bleeding index and probing depth (r = -0.54 and r = -0.65 respectively, P < 0.01). In the majority of periodontitis sites (91%), the concentration of IL-8 in GCF were lower than 30 microliters/L. IL-8 most likely acted as pro-inflammatory factor in these teeth. 2. IL-8 was a two-way regulator of inflammation, pro-inflammation and anti-inflammation. The threshold of IL-8 between inducing and suppressing inflammation was approximately 30 micrograms/L. Within the confined range, IL-8 concentration greater than 30 micrograms/L could be inflammation suppressive, while a less-than-30 micrograms/L concentration of IL-8 might become inflammation inducing. PMID- 10680521 TI - [Evaluation of reproducibilty of the hinge-axis and it's positional relation to condyles]. AB - The aim of this paper was to study the reproducibility of the positions of hinge axis of mandibular retruded small open-closed movements, and the positional relationship between the hinge-axis and condyles. Fourteen normal students were tested by a six degrees of freedom mandibular movement recorder MT1602 (Germany). The result showed that: the hinge-axis could not be exactly repeated and movable instantaneous centers of rotation (ICR) were recorded. These rotation centers distributed almost evenly around the condyles, the mean distance from these rotation centers to condyle centers was about 8.83 +/- 5.65 mm. PMID- 10680522 TI - [A comparative study of lanthanum, cerium and fluoride on the prevention of root surface caries by various procedure]. AB - In order to evaluate the anticaries effects of several trace elements by combined and respective treating procedure, studies on inhibiting demineralization and improving remineralization were carried out with lanthanum, cerium and fluoride in a pH-cycling condition. The analysis of polarized microscope and electron microprobe demonstrated that the lanthanum and fluoride to an acetate buffer solutions had less severe demineralization of root surface than individual lanthanum or fluoride to the solution. The former, furthermore, precipitated a compound membrane which may be F-La(Ce)-Ca containing crystal on the root surface, and showed a better resistibility to acid. A further discussion was made from the viewpoint of crystal physiochemistry. PMID- 10680523 TI - [The healing process of the temporalis myofascial flap in oral reconstruction]. AB - From a prospective study of 36 patients having had temporalis myofascial flap (mean follow-up 32 months), the clinical oral healing process and the histological characteristics of the repaired mucosa were investigated. Among this group, 24 patients were assessed of the clinical scarring over the reconstructed area and 11 patients for histological evaluation of the repaired mucosa. Result showed that the uncovered temporalis myofascial flap in the mouth underwent a consistent healing commencing with an acute inflammatory phase, through chronic inflammatory and proliferative phases to eventual epithelialization by oral mucosa. No major complications affecting the healing flap were noted. The healed mucosa exhibited scarring in 70% of cases and the repaired mucosa demonstrated histological features distinct from the normal mucosa. PMID- 10680524 TI - [Preliminary study of anatomic relation among nasopalatine duct, central incisor root and floor of nasal cavity with X-ray measurement]. AB - The authors studied anatomic constructure of anaterior maxilla. They observed the position relationships among nasopalatine duct, central incisor root and floor of nasal cavity through X-ray measurement. The results showed: (1) Average distance between the anterior wall of nasopalatine duct and cortical plate of anterior maxilla was 7.80 +/- 1.43 mm. (2) Average distance between the anterior wall of nasopalatine duct and central incisor root was 4.15 +/- 1.48 mm. (3) Average distance between the tip of central incisor root and floor of nasal cavity was 12.62 +/- 2.47 mm. (4) Average length of the nasopalatine duct was 15.04 +/- 2.20 mm. The results are beneficial of dental implantation and orthognathic surgery. PMID- 10680525 TI - [Observation of cervical vertebrae and estimation of their bone age]. AB - There are two objectives in this study: the first is to estimate skeletal age by lateral cephalomatric roentgengram of cervical vertebrae instead of X-ray of handwrist, the second is to study the rules of cervical vertebrae's growth and development of children from Beijing. The Auto CAD 12.0 computer software was used in measuring lateral cephalomatric roentgengrams of cervical vertebrae of 280 children from Beijing aged 9-15. The shape of cervical vertebrae of children with that of adults on X-ray films was compared, and the growth and development of cervical vertebrae of 9-15 years old children from Beijing was observed. We found out that the rapid growth period of cervical vertebrae was 12-14 years old for girls and 14-15 years old for boys. During puberty, the change of vertebrae's shape has no difference between male and female. 42 female and 28 male teenagers from the 280 aged 9-13 years old were taken X-ray films of left handwrist. The comparison between the films and roentgengrams shows that the appearance of sesamoid of hand and the concavity of the second vertebrae body is at the same time, which means that the beginning of rapid growth period can be estimated by the lateral cephalometric roentgengrams of cervical vertebrae. PMID- 10680526 TI - [Preliminary research on radiographic manifestations of health odontal and periodontal tissues in elderly people]. AB - Radiographs of mandibular first molars in 45 patients aged more than 60 year old were measured on several parameters of pulpal chamber, root canal as well as alveolar crest, and compared with those in 40 young people in order to study the radiographic manifestations of the healthy odontal and periodontal tissues in elderly people. Results shown that in aged people the average width and height of the pulpal chamber and the diameter of root canal were narrower; the root canal walls were thicker; the relation between the height of alveolar crest and the root length reduced significantly; and the lamina dura on the alveolar crest disappeared more common. Findings lay a preliminary foundation for investigation on radiographic manifestations of involved teeth in gerontology. PMID- 10680527 TI - [The immunogenicity of tooth allografts in mice]. AB - The mice used in this study were of the in-bred strains C57BL/6 and Balb/c which differ at the major histocompatibility complex (MHC). The immunogenicity of tooth allografts was studied in vitro by means of detecting the response of Antibody Complement Mediated Cytotoxity (ACMC) and Cell Mediated Cytotoxity (CMC) of the host. We found that there was significant difference for both ACMC and CMC activity between the allogenic and isogenic tooth transplantation (P < 0.01). The results indicate that tooth allografts are immunogens and do evoke the immune response of recipients. PMID- 10680528 TI - [A study on the changes of the estrogen level in the condylar cartilages in young growing rats after functional mandibular protrusion]. AB - Fifty female SD rats of 4-week-old were selected, divided equally and randomly into experimental and control groups. Simulated functional appliances were used to guide the mandibles of the rats forwards. The animals were sacrificed after 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks. The method of radioimmunoassay (RIA) was performed to quantitively detect the level of estrogen in the condylar cartilages. The results showed: 1. High levels of estrogen were detected in the condylar cartilages during their actively proliferative period. When they differentiated gradually, the levels of estrogen decreased. 2. The amount of estrogen was increased significantly after functional mandibular protrusion, especially during the period of the active proliferation of the condylar cartilages. It is suggested that estrogen has a close relationship with the hyperplasia and hypertrophy of the condylar chondrocytes and the increment of the levels of estrogen in the condylar cartilages will further promote the proliferation and functional adaptive remodelling of the condylar cartilages. PMID- 10680529 TI - [The change of calcitonin gene-related peptide positive fibers in inflammatory pulps]. AB - Deep cavities were prepared on rats' molars, and fresh carious dentin was enclosed in to establish animal pulpitis model. Immunohistochemical method was used to observe the distribution of calcitonin gene-related peptide (CGRP) positive nerve fibers in rats' normal dental pulps and inflammatory pulps at 4d, 8d, 14d respectively. CGRP positive fibers were found throughout the normal pulps, a large number of fibers penetrated into dentinal tubules. In 4d group, CGRP positive fibers became denser and the number of fibers penetrating into dentinal tubules also increased. In 8d group, a necrotic zone could be found in the coronal pulps, lots of fibers accumulated around the necrotic zone. In the radical pulps, CGRP positive fibers formed multiple arborizations. 14d postinflammation, all the pulps were necrotic, and the periapexs were richly innervated by CGRP positive fibers. The results suggesting that CGRP may play a role in the inflammation and repair of dental pulp. PMID- 10680530 TI - [Anatomy of the nasal cartilages of the unilateral lip and palate cleft nose]. AB - Noses of 6 stillborn infants with unilateral lip and complete palate cleft were dissected and analysed. The surgical dissection revealed that the lower lateral cartilages are asymmetrical on both sides, indicating displacement of the lower lateral cartilages and the cartilageous septum which deviates to the normal side. Paired T Test shown that there is no significant difference between two sides in length and width. The conclusion is that the nasal deformities of the lip and palate cleft are congenital and one of the major causal factors is the displacement of lower lateral cartilages and cartilageous septum in the cleft side. PMID- 10680531 TI - [Ameloblastic carcinoma and malignant amelobastoma: histological reunderstanding]. AB - Twenty-four cases of malignant ameloblastoma were reported, among them were nineteen cases followed up for 5 to 30 years. Histologic characteristics include predominated proliferation of the peripheral cells with papillary protruding to stroma, replacement of the stellate reticular cells in the follicular center by sarcomatous cells, as well as large and deep-stained or alveolar nuclei with moderate atypia, and high mitosis of the tumor cells. Clinico-radiologically, the tumor showed malignant features such as fast growing recently, pain, anesthesia, cauliflower ulcer, limited mouth opening, and large multilocular radiolucent area with ill-defined border. The biological behavior, nomenclature of the ameloblastic carcinoma or malignant ameloblastoma, and differential diagnosis from other tumors were also discussed. PMID- 10680532 TI - [Clear cell odontogenic carcinoma]. AB - Histopathologic and immunohistochemical features of 3 cases of clear cell odontogenic carcinoma were studied. The tumors were composed of sheets or islands of clear cells separated by mature collageneous tissue. Basaloid cells were also seen in the tumors. Tumor cells showed positive reactions for epithelial membrane antigen and cytokeratin. More PCNA positive cells were seen in basaloid cells than in clear cells. Metastasis to regional lymph nodes was found in all 3 cases. PMID- 10680533 TI - [Immunobiological significance of S-100 protein positive dendritic cells (S 100+DC) in patients with oral squamous cell carcinoma]. AB - The purpose of this study is to investigate the role and clinical significance of S-100+DC in oral squamous cell carcinoma. Distribution of S-100+DC in the tumor tissues and regional lymph nodes (LN) of 60 patients with oral squamous cell carcinoma was detected by immunohistochemistry (ABC). The results showed that the S-100+DC density in tumor tissues was correlated with the tumor histologic grade (P < 0.05). The density of S-100+DC was significantly higher in regional LN without tumor than those with metastases (P < 0.05). However, there was no difference of S-100+DC in tumor tissues between patients with and without regional LN metastases. The distribution of S-100+DC in tumor tissues and regional LN could be considered as a reference indicator of tumor histologic grade and clinical prognosis of patients. PMID- 10680534 TI - [The expression of proliferating cell nuclear antigen and p53 protein in salivary gland tumours]. AB - The expressions and distributions of PCNA and p53 protein were studied with anti PCNA and anti-p53 protein MoAbs, using immunohistochemical staining method in salivary gland tumour. The relationship between PCNA and p53 protein was also studied. Results indicated: Malignant mixed tumour (MmT) had a higher proliferating index (PI) of PCNA. The PI could be used as one of adjuvant diagnostic criteria; and as an important parameter for grading mucoepidermoid carcinoma (MEC); The positive expression of p53 might be a good tumour marker for MmT; Tumour tissues with positive expression of p53 had been shown higher PI of PCNA, however there was no significance difference in our group; It remained to be studied later. PMID- 10680535 TI - [Oral Candida and its pathogenesis]. PMID- 10680536 TI - [Clinical significance of temporomandibular joint disk displacement]. PMID- 10680537 TI - [Trends of the development of modern ophthalmology and the knowledge composition possessed by the ophthalmologists during the turn of centuries]. PMID- 10680538 TI - [The expression of EGF mRNA and EGF receptors in human trabecular meshwork cells in vitro]. AB - OBJECTIVE: To demonstrate that cultured trabecular meshwork cells can secrete epidermal growth factor (EGF) and there are EGF receptors (EGFRs) on the cells. METHODS: Human trabecular meshwork cells were cultured in vitro and passaged 3-5 times. Immunohistochemical stain was used to detect EGFRs on the trabecular meshwork cell membrane. EGF cDNA probe, alpha-32P isotope labeling and dot blot hybridization autoradiographic method were used to detect EGF mRNA in the cells. RESULTS: Cultured human trabecular meshwork cells were obtained. EGFR immunostain was positive, and the reaction of brown color was at the surface of the cells. Dot blot hybridization autoradiography showed that trabecular cells can express EGF mRNA. CONCLUSIONS: Trabecular meshwork cells can secrete EGF. There are EGFRs on the membrane of trabecular meshwork cells. It is suggested that probably up regulating the receptors or promoting the cells to secrete growth factors have potential significance in the regeneration of trabecular cells and recovery of the cell functions in open angle glaucoma. PMID- 10680539 TI - [The effects of pressure on cultured bovine trabecular meshwork cells]. AB - OBJECTIVE: To observe the effects of pressure on trabecular meshwork cells. METHODS: Bovine trabecular meshwork cells were cultured and submitted to different amounts of hydrostatic pressure. Cellular morphology and phagocytic function were observed under inverted phase-contrast microscope, light microscope and electron microscope. RESULTS: Compared with the control group, the cells under 2.0 kPa or 2.67 kPa for 48 hours had no remarkable difference in criteria observed. Those under 4.0 kPa for 24 hours showed slight changes in structure and a mild decrease in phagocytic function. The damage appeared more severe if the pressure was higher or lasted longer. CONCLUSION: Trabecular meshwork cells can only bear pressure below a certain level. They may be destroyed structurally or impaired functionally by pressure over this level. PMID- 10680540 TI - [Application of proliferating cell nuclear antigen in the study of human trabecular cell proliferation]. AB - OBJECTIVE: To evaluate the application of proliferating cell nuclear antigen (PCNA) in the study of human trabecular cell proliferation. METHODS: Immunohistochemical technique was used to observe the PCNA expressive level in the fourth passage of human trabecular meshwork, the effects of different concentrations of epinephrine, dexamethasone and epidermal growth factor on the level were also investigated, the results were compared with that of the normal cells cultured at the same time, and analyzed by a graphic device operation system in a computer. RESULTS: A stable proliferating curve was obtained according to the normal cell PCNA level, by which we could choose the best opportunity of drug application. Epinephrine and dexamethasone were found to significantly inhibit cell proliferation, while epidermal growth factor (EGF) could promote the proliferation. CONCLUSION: PCNA is considered to be a useful agent to observe the process of cell proliferation. The above mentioned methods are beneficial to the investigations of biochemical characteristics of trabecular meshwork cells and the pathogenic mechanisms of open-angle glaucoma. PMID- 10680541 TI - [Ahmed valve implantation for refractory glaucoma]. AB - OBJECTIVE: To evaluate the short-term and long-term efficacy of Ahmed valve implantation for refractory glaucoma and discuss its intraoperative and postoperative complications and management. METHODS: 20 cases of refractory glaucoma underwent Ahmed valve implantation, including 9 cases of neovascular glaucoma, 7 cases of aphakic or pseudophakic glaucoma, 3 cases of congenital glaucoma and one primary chronic angle-closure glaucoma. RESULTS: One month postoperatively, the intraocular pressure (IOP) was less than or equal to 2.8 kPa (1 kPa = 7.5 mmHg) in fifteen cases, with a total success rate of 75%. For cases with neovascular glaucoma, the success rate was 55.6%, while for the others, the rate was 90.9%. After six months of follow-up for 12 patients, the total success rate was 58.3%, for neovascular glaucomas and other glaucomas, the rate was 40.0% and 71.4%, respectively. The complications included transient hyphema, early postoperative hypotony, obstruction of the tube tip, tube touch to lens or cornea, tube exposure, exudative choroidal detachment and dropout of the plate. CONCLUSION: Ahmed valve implantation is an effective method in the management of refractory glaucoma in spite of its unnegligible complications. PMID- 10680542 TI - [Ocular echographic evaluation of human aqueous drainage implant]. AB - OBJECTIVE: To determine after implantation of a glaucoma aqueous drainage device whether fluid is present around the equatorial plate and to measure the bleb size around the plate. METHODS: 41 plates in 41 patients (41 eyes) with refractory glaucoma who had undergone human aqueous drainage (HAD) implantation underwent standardized echography (both A- and B-scan examinations) after a mean postoperative follow-up period, 5.2 months. According to the bleb height and the fluid underlying and/or overlying the plate, the blebs were divided into five grades and two types to evaluate their functions. RESULTS: Of the 41 cases examined with echography, 35 (85%) were associated with posterior blebs, 2 (5%) had no associated posterior blebs, and the posterior bleb in 4 (10%) cases was not certain. The percentage of the presence of filtering bleb around the plate was 15%, 12%, 29%, 37%, and 7% in Grade 0 to IV group, and 11% and 89% in type A and B, respectively. There was no significant correlation between the bleb size and the level of intraocular pressure control. CONCLUSION: Standardized ocular echography is helpful in the postoperative management of failed cases who have undergone HAD implantation because it can demonstrate the presence or absence of blebs and characterize them. PMID- 10680543 TI - [The relation of the changes of SOD and PGE2 in aqueous humor with the transient rise of intraocular pressure after argon laser iridectomy]. AB - OBJECTIVE: To study the mechanism of transient rise of the intraocular pressure (IOP) after argon laser iridectomy (ALI). METHODS: The rabbit changes of IOP, the activity of superoxide dismutase (SOD) and the concentration of prostaglandin E2 (PGE2) in aqueous 15 min, 60 min, 6 hr, 24 hr, 48 hr after ALI, the relationships of these changes with the transient rise of IOP, and the effects of using indomethacin, cortisone, vitamin E and C before ALI on the transient rise of IOP were observed. RESULTS: After ALI, the transient rise of IOP took place within postoperative 6 hr. Following the postoperative transient rise of IOP, the concentration of PGE2 in the aqueous was markedly increased, but the activity of SOD in the aqueous markedly decreased. When indomethacin, cortisone, vitamin E and C were used before ALI, the concentration of PGE2 was not markedly increased, the activity of SOD was not markedly decreased and the postoperative transient rise of IOP did not take place. CONCLUSIONS: The transient rise of IOP after ALI is related to the increase of concentration of PGE2 and the decrease of activity of SOD in the aqueous. Using indomethacin, cortisone, vitamin E and C before ALI can effectively prevent the postoperative transient rise of IOP. PMID- 10680544 TI - [Protective effects of exogenous superoxide dismutase on rabbit retinal injury by acute intraocular hypertension]. AB - OBJECTIVE: To study active oxygen and free radical injury in rabbit retina during elevated intraocular pressure and the protective effect of exogenous superoxide dismutase (SOD) on the retinal damage by the hypertension. METHODS: Lipid peroxidative product, malondialdehyde (MDA), activity of SOD and glutathione peroxidase (GSH-Px), reduced GSH in the retinal tissue were measured during 24 h after the release of an ocular hypertension, 6.67 kPa (1 kPa = 7.5 mmHg) maintaining for 1.5 h, and the effects of retrobulbarly injected Cu-Zn-SOD on the level of MDA and the activity of SOD in the retinal tissue after the release of ocular hypertension for 12 h were observed. RESULTS: MDA increased gradually during 0-12 h after the release of ocular hypertension and maintained at a relatively high level in 12-24 h. The activity of SOD and GSH-Px was lower than normal level immediately after the release, and then increased to a certain different extent. But the activity of SOD began to decrease gradually 4 h after the release. GSH had no significant changes during 24 h after the release. Retrobulbar injection of Cu-Zn-SOD reduced the production of MDA in the retinal tissue and enhanced SOD activity. CONCLUSIONS: Active oxygen and free radicals participate the rabbit retinal injury by elevated intraocular pressure. A high dose of Cu-Zn-SOD retrobulbar injection plays a beneficial role in enhancing the antioxidative ability of the retina. PMID- 10680545 TI - [Quantitative analysis of cAMP and cGMP in the plasma, iris and aqueous humor in rabbits with high intraocular pressure]. AB - OBJECTIVE: To study the relationship between the levels of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophos-phate (cGMP) in the plasma, iris and aqueous humor and high intraocular pressure in rabbits. METHODS: The quantitative tests of cAMP and cGMP from rabbits were carried out by radioimmunoassay technique. RESULTS: The results showed that there were 72.81 +/- 7.67, 141.08 +/- 16.70 and 28.40 +/- 0.49 of cAMP (nmol/L), and there were 10.37 +/- 6.15, 35.64 +/- 7.02 and 12.90 +/- 0.61 of cGMP (nmol/L) in the plasma, iris and aqueous humor respectively. The levels of cAMP were higher than those in the same samples from the animals as controls (P < 0.01), while the levels of cGMP were less than controls (P < 0.01). CONCLUSION: High intraocular pressure may be significantly associated with the levels of cAMP and cGMP. PMID- 10680546 TI - [Changes of corneal topography and refraction after radial keratotomy and its combination with transverse incision]. AB - OBJECTIVE: In order to understand the changes of morphology and refractive power of cornea after radial keratotomy (RK) and its combination with transverse incision (T-incision), and the wound healing of the T-incision. METHODS: Corneal topography aided with a computer was used to analyze the changes of patients having undergone RK and RK combined with T-incision. The wound healing was analyzed, and the refractive powers of strong axis and weak axis of corneas were measured. RESULTS: Corneal surface regularity index (SRI) and surface asymmetry index (SAI) became greater and greater along with the increase of myopia astigmatism. The refractive powers of corneal visual area 3, 5, 7 mm decreased when getting closer to the center of the cornea. The refractive power of the strong axis in cornea with RK combined with T-incision obviously decreased more than that of RK. There was no such difference at weak axis between RK and RK combined with T-incision. CONCLUSIONS: Corneal SAI and SRI will increase when myopia-astigmatism increases. The morphologic changes are from positive non spherical lens before surgery to negative non-spherical lens after surgery. The increase of radius of curvature and the decrease of refractive power are due to incisions. These phenomena become clearer when the incision getting closer to the visual area or more incisions are performed. A T-incision less than 2 mm long along the strong axis of astigmatism not only can lessen refractive power of strong axis, but also can not affect RK results. This kind of T-incision does not delay the wound healing either. PMID- 10680547 TI - [A study of multi-channel visual evoked potentials in exotropic children without amblyopia]. AB - OBJECTIVE: To investigate the clinical significance of multi-channel visual evoked potentials (VEPs) in constant exotropic children without amblyopia and the mechanism of exotropia. METHODS: With 14 active cup electrodes, and full and half field stimulation, the checkerboard pattern was reversed and the VEPs were recorded in control and experimental groups. RESULTS: With full field stimulation, the latent period 1 (LP1) of binocular VEPs was not longer and the amplitude N1-P1 (AN1P1) not higher respectively than those of monocular VEPs in experimental group. These properties were different from those of control group. The AN1P1 of non-dominant eye was decreased, and the LP1 of dominant and non dominant eye was longer in experimental group than that of the control group. There were no significant differences in comparing LP1 and AN1P1 between nasal retina and temporal-retina of experimental group. CONCLUSIONS: VEPs can provide a scientific basis for abnormal binocular vision in constant exotropia. Although the visual acuity of constant exotropic children is normal, their VEPs are abnormal. Therefore, there might be dysfunction in primary visual cortex. With half field stimulation, there is no suppression of temporal-retina in monocular VEPs of constant exotropic children. PMID- 10680548 TI - [Clinical observation on pars plana lensectomy-vitrectomy preserving lens anterior capsule]. AB - OBJECTIVE: To investigate the therapeutic effect of combined operation on cataract complicated with vitreous pathology. METHOD: 19 patients (20 eyes) with coexisting cataract and vitreous opacification or endophthalmitis or intraocular foreign bodies were operated with pars plana lensectomy-vitrectomy preserving lens anterior capsule. The ciliary sulcus intraocular lenses (IOLs) were primarily implanted for sixteen eyes of 19 cases in the operation. RESULTS: All cases were followed up for 12-36 months. Visual acuities of 18 cases were improved, 7 eyes between 0.3-0.5, 11 eyes beyond 0.5, the best one at 1.2. Anterior capsule opacification occurred merely in one eye, and in others the capsule kept being transparent. CONCLUSION: Preserving anterior lens capsule in the operation is beneficial to the insertion of IOL, and the anterior segment is affected slightly, thus it is a promising method for treating cataract complicated with vitreous opacification. PMID- 10680549 TI - [Diagnosis and treatment of blowout fracture of the orbit]. AB - OBJECTIVE: To analyze the diagnosis and treatment of blowout fracture of the orbit. METHODS: 78 cases with blowout fracture of the orbit seen in 20 years were retrospectively analyzed. In different years, different methods of examinations were used. There were 16 cases who had undertaken X-ray examinations, 31 cases, ultrasonographic examinations and 72 cases, CT. RESULTS: The positive rate of the CT examinations with blowout fracture of the orbit was 100%, CT having the tendency to eventually replace the X-ray. Ultrasonography shows only changes of intraorbital tissue, providing an auxiliary method for diagnosis. CONCLUSION: With the progress of modern photographic methods, blowout fracture of the orbit is known more deeply and the advance helps in the improvement of the treatment of the disease. PMID- 10680550 TI - [Cytological study of subretinal fluid]. AB - OBJECTIVES: To determine the cellular components of subretinal fluid and the origin of cells participating in the proliferation in the membranous tissue of proliferative vitreoretinopathy (PVR). METHODS: 35 subretinal fluid samples were studied by cytology using autofluorescent and immunohistochemical assays. RESULTS: There were a large quantity of retinal pigment epithelium (RPE) and small quantity of pigment epithelial cells originated from epithelial cells of ciliary body and iris, there were also various quantities of neuroretinal cells and macrophages. Neither T-lymphocytes nor B-lymphocytes were found in this group of samples. CONCLUSIONS: RPE, pigment epithelial cells, neuroretinal cells and macrophages are in subretinal fluid and may participate in the formation and development of PVR in various degrees. PMID- 10680551 TI - [A preliminary study on the association between HLA-DPB1 gene and pathological myopia]. AB - OBJECTIVE: To investigate the association between distribution of human leucocyte antigen class II DPB1 alleles and pathological myopia (PM) among Chinese. METHODS: The second exons of the HLA-DPB1 genes of 40 patients with PM were amplified by polymerase chain reaction (PCR), individual PCR products were digested by allele specific restriction enzymes: Bsp 1296 I, Fok I, Dde I, BsaJ I, BssH II, Rsa I, Ava II and EcoN I. Genotype of each patient was determined by restriction fragment length polymorphism (RFLP) pattern, then each DPB1 allele frequency of PM was calculated and compared with healthy control. RESULTS: The frequency of DPB1*0301 allele was significantly decreased in PM (Yates corrected chi 2 = 4.33, Fisher exacted P = 0.032 < 0.05) and no statistically significant difference was observed after P value was corrected by the number of compared alleles. The rate of DPB1*0501/0501 homozygote has a significant difference between PM and healthy controls (u = 2.27, P < 0.05). CONCLUSIONS: No genetic susceptible or resistant allele exists in HLA-DPB1 gene of PM. The significant increase of DPB1*0501/0501 homozygote in PM may be a linkage information. PMID- 10680552 TI - [Research of daunomycin as an agent for preventing posterior capsule opacification]. AB - OBJECTIVE: To investigate the inhibitory activity and the effective concentration of daunomycin against the proliferation of lens epithelial cells in vitro, and make progress in pharmacological prevention of posterior capsule opacification. METHODS: Cultured bovine, rabbit and human lens epithelial cells were exposed to 0.5, 2.5, 5.0, 7.5 and 10.0 micrograms/ml daunomycin solution for ten minutes. Growth of cells was observed, after 48 hours of exposure, absorbency of the cells was tested by using Giemsa staining and colorimetry, and the median lethal dose (LD50) was calculated respectively. RESULTS: Daunomycin significantly suppressed the proliferation of bovine, rabbit and human lens epithelial cells in vitro, showing dose-dependent manner. Daunomycin can effectively inhibit the proliferation of human lens epithelial cells at the concentration of 0.5 microgram/ml, almost reaching the greatest effect at the concentration of 7.5 micrograms/ml. The calculated LD50 for bovine, rabbit and human lens epithelial cells was 0.49, 4.30 and 4.06 micrograms/ml respectively. CONCLUSION: Short time exposure of daunomycin at low concentration can effectively inhibit the proliferation of lens epithelial cells in vitro, after further research in vivo, daunomycin may possibly become a valuable agent for the prevention of posterior capsule opacification. PMID- 10680553 TI - Sex effects on rate of change of P300 latency with age. AB - OBJECTIVE: Recent MRI evidence suggests that neuroanatomic structures may change more rapidly with age in males compared with females. Sex differences for P300 latency were tested to determine whether similar results might appear for P300 latency, a neurophysiological measure sensitive to age and neurodegenerative processes. METHODS: Auditory event related potentials (ERPs) were recorded using an auditory 'oddball' to elicit the N200 and P300 components. Forty-two male and 42 female healthy normal subjects (age range 15-85 years) were entered in this study. Both linear and curvilinear correlations of N200 and P300 latency/amplitude with age were tested. RESULTS: The slope of P300 latency on age for males was steeper than for females at Pz in subjects who were 30 years of age and older. N200 and P300 latencies were inversely correlated with age in young adult males (<30 years old). CONCLUSIONS: Males may experience more rapid change of P300 latency, but not amplitude, than females in middle to old age. Further research is required to determine whether those change reflects neural pathophysiology, or is mediated by such factors as neuroanatomic differences, body temperature, or mild auditory deficits. PMID- 10680554 TI - Event-related brain potentials in response to novel sounds in dementia. AB - OBJECTIVE: Non-target, deviant stimuli generate an earlier latency, front-central novelty P3, whereas correctly detected task-relevant stimuli generate a parietal maximal target P3. We examined whether the P3 component to novel stimuli is affected by dementing processes, and is therefore useful for distinguishing Alzheimer's type dementia (AD) from vascular dementia (VD). METHODS: We recorded ERPs to task-relevant stimuli (target P3) and novel task-irrelevant stimuli (novelty P3) in an auditory oddball task in AD (n = 16), VD (n = 16), and age matched controls (n = 18). The amplitude, latency, and scalp topography of target and novelty P3 were compared among 3 groups using ANOVA. The relationship between P3 measures and intelligence scores were evaluated by correlation analysis. RESULTS: The amplitude, latency and scalp topography of the target P3 were comparably affected by both AD and VD. However, the amplitude of the novelty P3 was markedly reduced in VD, but not in AD, and the scalp topographics were different in the 3 groups. The amplitude was maximal at frontal sites in controls, at central sites in AD, and at parietal sites in VD. The target P3 latency was prolonged in both AD and VD, whereas the novelty P3 latency was only prolonged in VD. AD was discriminated satisfactorily from VD by using the novelty amplitude at Cz and the ratio of the amplitudes at Fz and Pz as independent variables. CONCLUSIONS: These results suggest that the response to novel stimuli is differentially affected by dementia with degenerative and vascular etiology. PMID- 10680555 TI - Sequential information processing during a mental arithmetic is reflected in the time course of event-related brain potentials. AB - OBJECTIVE: We hypothesized that mental arithmetic is a complex of sequential information processing. In order to test the hypothesis, event-related potentials (ERPs) were measured during 3 mental tasks. METHODS: Fifteen normal human subjects performed the following tasks; (1) subjects added up every digit delivered successively on a computer display, (2) subjects counted the number of presented digits, or (3) subjects counted the number of meaningless patterns. Spatiotemporal differences between ERP waveforms under the 3 tasks were studied within the period of 1200 ms post-stimulus. RESULTS: During the adding task, N120 P180-N220 complex advanced in latency in the left frontal, central and parietal regions. P300 increased in amplitude in the frontal and temporal regions during adding and counting digits, which was specific to the digit presentation. A positive slow potential depended on the adding task and showed two spatiotemporal distributions; one was a widespread brain activity over the frontal, central, temporal and parietal regions observed within 400-820 ms, and another was a brain activity restricted to the frontal region lasting up to 1150 ms. CONCLUSIONS: The results suggest that the early portion of ERPs reflects identification of physical attributes of stimuli and numeric meaning of digits, and that the positive slow potential reflects processes associated with calculation. PMID- 10680556 TI - Identification of auditory evoked potentials of one's own voice. AB - OBJECTIVE: We recorded vocalization related cortical potentials (VRCP) with complete masking of one's own voice, and separated the feedback auditory potentials following vocalization from the VRCP complex. METHODS: We used 9 right handed healthy subjects. We recorded VRCP during simple vowel vocalizing under two conditions, (1) without masking (control); (2) with masking. Electroencephalography (EEG) was recorded from seven areas, Fz, Cz, Pz, C3, C4, T3 and T4, of the International 10-20 system. The trigger point was the onset of vocalized sound recorded through a microphone. RESULTS: In both control and masking conditions, similar negative potentials prior to the vocalization were observed. However, the amplitude just after the onset of vocalization was significantly smaller in the masking than the control condition. The difference waveform, obtained by subtracting the waveforms in the masking from those in the control condition, showed a simple negative peak after the vocalization onset. CONCLUSIONS: We considered that the difference waveform indicated the auditory evoked potentials following own voice in the auditory feedback process. Since there was no difference of the negative potentials before vocalization between the two conditions, we speculated that the masking effect on auditory feedback following vocalization was crucial, while its effect on the preparatory process of simple vocalization might be minimal. PMID- 10680557 TI - Maturation of human central auditory system activity: evidence from multi-channel evoked potentials. AB - OBJECTIVE: The purpose of this study was to evaluate central auditory system maturation based on detailed data from multi-electrode recordings of long-latency auditory evoked potentials (AEPs). METHODS: AEPs were measured at 30 scalp electrode locations from 118 subjects between 5 and 20 years of age. Analyses focused on age-related latency and amplitude changes in the P1, N1b, P2, and N2 peaks of the AEPs generated by a brief train of clicks presented to the left ear. RESULTS: Substantial and unexpected changes that extend well into adolescence were found for both the amplitude and latency of the AEP components. While the maturational changes in latency followed a pattern of gradual change, amplitude changes tended to be more abrupt and step-like. Age-related latency decreases were largest for the P1 and N1b peaks. In contrast, P2 latency did not change significantly and the N2 peak increased in latency as a function of age. Abrupt changes in P1, P1-N1b, and N2 peak amplitude (also RMS amplitude) were observed around age 10 at the lateral electrode locations C3 and C4, but not at the midline electrodes Cz and Fz. These changes in amplitude coincided with a sharp increase and plateau in AEP peak and RMS amplitude variability from 9 to 11 years of age. CONCLUSIONS: These analyses demonstrated that the observed pattern of AEP maturation depends on the scalp location at which the responses are recorded. The distinct maturational time courses observed for individual AEP peaks support a model of AEP generation in which activity originates from two or more at least partly independent central nervous system pathways. A striking parallel was observed between previously reported maturational changes in auditory cortex synaptic density and, in particular, the age-related changes in P1 amplitude. The results indicate that some areas of the brain activated by sound stimulation have a maturational time course that extends into adolescence. Maturation of certain auditory processing skills such as speech recognition in noise also has a prolonged time course. This raises the possibility that the emergence of adult like auditory processing skills may be governed by the same maturing neural processes that affect AEP latency and amplitude. PMID- 10680558 TI - The effect of interstimulus intervals and between-block rests on the auditory evoked potential and magnetic field: is the auditory P50 in humans an overlapping potential? AB - OBJECTIVE: The human auditory P50 may consist of overlapping potentials. To test this hypothesis, we manipulated interstimulus intervals (ISIs) and between-block rests, and recorded the P50, P50m, N100 and the N100m simultaneously. METHODS: Subjects were 12 right-handed healthy adults. Four conditions included: (1) 1.5 s/rest, (2) 1.5 s/successive, (3) 0.5 s/rest, and (4) 0.5 s/successive. Auditory stimuli were presented a total of 880 times for each condition. Auditory evoked potentials and magnetic fields were recorded. We examined the P50, N100, P50m, N100m and dipoles of the P50m and the N100m. RESULTS: There was no significant effect of the ISI on the P50 amplitudes, but the P50m amplitudes in the 0.5 s ISI conditions were significantly smaller than those in the 1.5 s ISI conditions. The N100 and the N100m amplitudes in the 0.5 s ISI conditions were significantly smaller than those in the 1.5 s ISI conditions. The N100 and the N100m amplitudes in the resting conditions were significantly larger than those in the successive conditions. The P50m dipoles were slightly deeper and more anterior than those of the N100m in primary auditory cortex. CONCLUSIONS: Central structures other than supratemporal cortex contribute to the P50 and that the P50 in humans represents overlapping potentials. PMID- 10680559 TI - Steady-state visual evoked potentials and travelling waves. AB - OBJECTIVE: The amplitude and phase of the steady-state visual evoked potential (SSVEP) is sensitive to cognition and attention but the underlying mechanism is not well understood. This study examines stimulus evoked changes in the SSVEP phase topography and the putative role of travelling waves. METHODS: Eighteen subjects viewed a central-field checkerboard and full-field flicker stimulus temporally modulated at the peak alpha rhythm frequency. EEG was recorded from 10 midline scalp sites and the bipolar SSVEP obtained from differences between adjacent electrodes. RESULTS: The SSVEP phase comprised either progressive variations consistent with travelling waves or a phase reversal consistent with standing waves. The checkerboard pattern elicited travelling wave patterns in 14 subjects with estimated phase velocities ranging from 7 to 11 m/s after correcting for folded cortex. The flicker stimulus elicited phase reversals in 9 subjects, suggesting standing waves. Six subjects demonstrated a phase topography specific to the stimulus with travelling wave patterns associated with the checkerboard and standing wave patterns associated with the flicker. CONCLUSIONS: These differences suggest the emergence of travelling and standing waves under different spatial configurations of visual input to the cortex and that wave phenomena contribute to the spatiotemporal dynamics of the SSVEP. PMID- 10680560 TI - Functional properties of sub-bands of oscillatory brain waves to pattern visual stimulation in man. AB - The scalp recorded transient visual evoked potential (VEP) represents the massed activity of a large number of neurons of the human visual cortex. Animal studies show that intracerebrally-recorded high frequency electrical activity represents binding between neurons participating in a cooperative response. We evaluated the relationship between scalp recorded high frequency activity and transient VEPs elicited by a repetitive (grating) pattern. Stimuli were 1 and 4 cycles/degree sinusoidal gratings, presented in an on/off mode. Following conventional averaging, the discrete wavelet transform (DWT) was applied. Multi-resolution decomposition was used to divide the responses into 6 orthogonal frequency bands. The results show that high frequency oscillatory activity in the beta and gamma frequency range is closely related in time to the N70 peak of the simultaneous VEP. Power in both bands is modulated by spatial frequency. Beta range response to hemifield stimulation recorded over a chain of electrodes over the occipital area lateralizes in the same manner as N70, while gamma range activity is insensitive to lateralization and is more closely linked to foveal stimulation. This dissociation between beta and gamma range activity suggests that different bands of high frequency oscillatory activity in humans, linked to visual stimulation, may represent different aspects of visual processing. PMID- 10680561 TI - Dysfunction of small myelinated afferents in diabetic polyneuropathy, as assessed by laser evoked potentials. AB - OBJECTIVE: To verify whether laser evoked potentials are useful in assessing the function of small afferent fibers and to compare dysfunction of large and small afferent fibers in patients with diabetic polyneuropathy. METHODS: The brain potentials evoked by CO2 laser stimulation of the hand and foot were studied in diabetic patients (n = 45) with various degrees of peripheral nerve damage. Laser evoked potentials (which assess the function of small myelinated afferents) were also compared with ulnar and sural nerve sensory action potentials (which assess the function of large myelinated afferents) by scoring the abnormalities of the two neurophysiological tests with similar criteria. RESULTS: Laser evoked potentials were often absent; the mean latency was normal and mean amplitude decreased, as expected in axonopathies. Although clinical examination showed more frequent impairment of vibratory than pinprick sensation, laser evoked potentials and sensory action potentials yielded similar abnormality scores and showed a strong intra-individual correlation. CONCLUSIONS: Laser evoked potentials, possibly better than standard clinical examination for assessing the abnormalities of small-diameter afferents, indicate that diabetic polyneuropathy induces large- and small-afferent dysfunction in parallel. PMID- 10680562 TI - Homeostatic sleep regulation in patients with idiopathic hypersomnia. AB - OBJECTIVE: To determine whether the spectral activity during sleep of patients with idiopathic hypersomnia (IH) differs from that of healthy subjects. METHODS: Spectral analysis of the electroencephalogram (EEG) was performed in 10 patients with IH and in 10 age-matched control subjects. We compared the time course of absolute power for slow wave activity (SWA: 0.75-4.5 Hz), and for theta, alpha, sigma and beta bands for the first 4 non-rapid-eye movement (NREM) episodes. RESULTS: Compared to controls, IH patients had less SWA across the night although the exponential decay was preserved. The fall in SWA was statistically significant for the first two NREM episodes only. The lower power of SWA was related to lower amounts of stages 3 and 4 of NREM sleep during the sleep episodes. No correlation was found between SWA during the night and the mean sleep latency on the multiple sleep latency test (MSLT). CONCLUSIONS: These results showed that, in IH patients, the homeostatic sleep regulatory mechanisms are preserved but the sleep pressure, indicated by SWA, is lower. PMID- 10680563 TI - CAP components and EEG synchronization in the first 3 sleep cycles. AB - OBJECTIVE: There is consolidated evidence that stage changes in sleep are closely related to spontaneous EEG fluctuations centered on the 20-40 periodicity of the cyclic alternating pattern (CAP). The present investigation aimed at assessing the involvement of the different components of CAP in the process of build-up, maintenance and demolition of deep non-REM (NREM) sleep. METHODS: CAP parameters were quantified in the first 3 sleep cycles (SC1, SC2, SC3), selected from polysomnographic recordings of 25 healthy sound sleepers belonging to an extensive age range (10-49 years). Only ideal SCs were selected, i.e. the ones uninterrupted by intervening wakefulness and in which all stages were represented and linked in a regular succession of a descending branch, a trough and an ascending branch. RESULTS: Among the first 3 SCs, a total amount of 45 (SC1, 16; SC2, 13; SC3, 16) met the inclusion requirements. SCI contained the highest amount of slow wave sleep (43.7 min) and the lowest values of CAP rate (31.6%). The number of phase A1 subtypes remained unmodified across the 3 SCs (SC1, 48; SC2, 48; SC3, 48), whereas both subtypes A2 (SC1, 9; SC2, 14; SC3, 14) and A3 (SC1, 2; SC2, 8; SC3, 10) increased significantly (P<0.028 and P<0.0001, respectively). The A1 subtypes composed more than 90% of all the A phases collected in the descending branches and in the troughs, while the A2 and A3 subtypes were the major representatives (64.3%) of the A phases occurring in the ascending branches. CONCLUSIONS: Within the dynamic organization of sleep, the non-random distribution of CAP sequences, with their succession of slow (subtypes A1) and rapid (subtypes A2 and A3) EEG shifts, seem to be responsible for sculpturing EEG synchrony under the driving and alternating forces of NREM and REM sleep. PMID- 10680564 TI - Measurement of lingual and palatine somatosensory evoked potentials. AB - OBJECTIVE: A technique is presented for generating and recording lingual and palatine nerve somatosensory evoked potentials (SEPs). METHODS: Pairs of thin, stainless steel disk electrodes were mounted onto mandibular or maxillary acrylic splints, similar to orthodontic retainers. Mandibular splint electrodes were oriented to contact the under surface of the tongue along the course of the right and left lingual nerves and maxillary splint electrodes were oriented to contact the hard palate bilaterally along the course of the palatine nerves. SEP recording electrodes were placed on the scalp 1 cm posterior to C5 and C6 (C5' and C6', respectively) using the combinatorial nomenclature of the International 10-20 system. Two reference electrode locations, Fz and C5' or C6', over the cortical hemisphere opposite that of the recording electrode, were used. RESULTS: Right and left lingual and palatine nerve SEPs were recorded from five normal adults. SEP latencies were similar to the N13 and P18 cortical peak latencies recorded in previous studies of trigeminal nerve branches to the lips regardless of reference electrode position. CONCLUSIONS: A more precise method of stimulating the intraoral lingual and palatine nerves was accomplished using dental splints. SEPs were easier to obtain using a contralateral cortex reference electrode location. PMID- 10680565 TI - Postanoxic alpha (theta) coma: a reappraisal of its prognostic significance. AB - OBJECTIVES: To appraise the controversial prognostic significance of postanoxic alpha or theta coma (ATC). METHODS: We prospectively assessed 14 comatose patients with ATC after cardiac arrest by means of a protocol which included repeated clinical examinations, EEG, and median somatosensory evoked potentials (SEP). Good outcome was defined by the reappearance of cognition (Glasgow outcome scale 3-5) at any time during the 1 year follow-up. RESULTS: Nine of 14 patients had a monotonous, frontally accentuated and a reactive alpha (theta) EEG activity (complete ATC). In these patients ATC was recorded a mean of 47 h after resuscitation, the mean Glasgow coma scale (GCS) was 4 at 48 h, and early cortical SEPs were altered or absent in 5 of 7 patients. All nine patients died. In five of 14 patients the alpha (theta) EEG activity was either not monotonous, partially reactive or posteriorly dominant (incomplete ATC). In these patients ATC was recorded a mean of 43 h after resuscitation, the mean GCS was 8 at 48 h, and early cortical SEP were normal in 4 of 5 patients. Three of 5 patients regained cognition, two of them remained however dependent in activities of everyday life. CONCLUSIONS: This study and a review 283 cases of postanoxic ATC reported in the literature suggest the existence of incomplete and complete variants of postanoxic ATC. Whereas complete ATC is invariably associated with a poor outcome, full recovery is possible in patients with incomplete ATC. The combination of EEG, clinical, and SEP findings improves the prognostic accuracy of postanoxic ATC. PMID- 10680566 TI - Differences in the dynamics of frontal sharp transients in normal and hypoglycemic newborns. AB - OBJECTIVE: This study focuses on the characteristics of frontal sharp transients (FST), a normal variant of neonatal EEG, in newborns with hypoglycemia. METHODS: The EEG from 20 newborns with symptomatic hypoglycemia were compared with a control group of normal newborns matched by conceptional age. The dynamics of these transients were evaluated concerning sleep stages. The density, type and bilateral synchrony of FST were also calculated. The results were compared in the two groups by Student's t test. RESULTS: When compared with controls, hypoglycemic newborns have increased density of frontal sharp transients in all sleep stages but less bilateral synchrony (P<0.05). FST density was even higher in small for gestational age newborns. CONCLUSIONS: The data suggests that normal patterns of neonatal EEG as FST can be influenced by systemic disturbances. PMID- 10680567 TI - Gabapentin as add-on therapy in focal epilepsy: a computerized EEG study. AB - OBJECTIVES: Gabapentin (GBP) possesses a well documented clinical efficacy in those types of focal epilepsy otherwise resistant to conventional antiepileptic drugs (AEDs); on the basis of this, it appears important to investigate the drug effects on the EEG epileptiform and background activity. METHODS: Twenty-five patients with cryptogenic or symptomatic partial epilepsy resistant to conventional AED treatment were included in the study. All patients underwent long-term video-EEG recordings before and after GBP addition (900-1200 mg/day). RESULTS: Quantitative analysis of the interictal EEG paroxysms revealed that GBP had no effect on the rate of occurrence of interictal and ictal EEG abnormalities. GBP was active in delimiting the spatial extent of the interictal spiking activity in those patients who displayed a significant reduction (> or =50%) in seizure occurrence (32% of the patients). EEG background activity recorded under rest condition from 18 out of 25 epileptic patients, before GBP therapy, was characterised by a higher content of the slow spectral components (delta and theta) with respect to control subjects. After GBP addition, the increase of theta relative power was also evident during task performance. CONCLUSIONS: These findings suggest that GBP does not interfere with the generation of interictal EEG spiking while it appears to reduce the susceptibility to seizures concomitantly with a limiting effect on the spiking activity spatial extent. The utilization of GBP in controlling focal seizures is reinforced by the absence of negative influence on cognitive functioning. PMID- 10680568 TI - Patterns of motor control reorganization in a patient with mirror movements. AB - OBJECTIVE: To explore motor control reorganization in a 40-year-old, left-handed patient with perinatally acquired mirror movements. METHODS: We performed simultaneous bilateral recordings of motor evoked potentials (MEPs) following focal transcranial magnetic stimulation (fTMS) and of central silent period (cSP) during unilateral voluntary contraction in abductor pollicis brevis (APB) and abductor digiti minimi (ADM) muscles. RESULTS: For both muscles the MEP study showed bilateral fast-conducting corticospinal projections from the right undamaged hemisphere, and residual contralateral projections from the left hemisphere. The cSP findings differed in the two muscles: the mirror phenomenon was bilateral in the ADM, but present only on the right side in the APB muscles; the mirror activity of right ADM and APB muscles was inhibited only by fTMS of the ipsilateral right motor cortex; the mirror phenomenon in the left ADM muscle was inhibited only by fTMS of the contralateral right motor cortex. CONCLUSIONS: Mirror movements of right APB and ADM muscles were sustained by the ipsilateral connections from the undamaged motor cortex, while the mirror phenomenon in the left ADM muscle could be explained by hypothesizing a bilateral activation of motor cortices. This previously unreported electrophysiological picture demonstrates that different patterns of motor control may realize after perinatal cerebral lesions, even in different distal muscles of the same patient. PMID- 10680569 TI - Electroencephalographic measurement of motor cortex control of muscle activity in humans. AB - OBJECTIVE: To detect and measure correlation between cortical and muscle activities, coherence analysis was used. METHODS: The electroencephalogram (EEG) and electromyogram (EMG) were recorded in 9 normal volunteers during tonic contraction of upper and lower limb muscles on the right side. Coherence between EEG and EMG was computed to analyze their linear association. RESULTS: EEG over the contralateral sensorimotor area was coherent with EMG, with peak coherence at 11-36 Hz (mean, 22 Hz). For the abductor pollicis brevis (APB) muscle, peak coherence, as determined by functional brain mapping with focal transcranial magnetic stimulation (TMS), was over or slightly posterior to the hand area on the primary motor cortex determined by focal transcranial magnetic stimulation (TMS). Peak coherence over the scalp was somatotopically organized. The temporal relation between EEG and EMG was analyzed with a new model for interpreting the phase shift ('constant phase shift plus constant time lag' model). For the APB muscle, the phase relation between cortical and muscular oscillations differed in the frequency ranges of 3-13 Hz and 14-50 Hz, respectively, suggesting that different coupling mechanisms operate in different bands. Only the phase shift between cortical and motoneuronal firing at 14-50 Hz was reliably estimated by a linear model. At 14-50 Hz, motoneuronal firing was led by surface-negative cortical activity with a constant time lag that depended on the cortical-muscular distance. For the APB muscle, the time lag was slightly shorter than the cortical muscular conduction time determined by TMS. Vibratory stimulation (100 Hz) of a muscle tendon during tonic contraction had no significant effect on cortical muscular coherence, indicating that cortical oscillation reflected motor rather than sensory activity. CONCLUSIONS: The present findings suggest temporal coding of the oscillatory motor control system (3-13 Hz vs. 14-50 Hz), and confirm the functional importance of cortical beta and gamma rhythms in the motor efferent command. Cortical-muscular synchronization is most likely mediated by the direct corticospinal pathway within the frequency range of 14-50 Hz. PMID- 10680570 TI - A novel quantitative method for 3D measurement of Parkinsonian tremor. AB - OBJECTIVE: To demonstrate the usefulness of a three dimensional (3D) motion analysis system for the quantitative measurement of tremor in patients with Parkinson's disease (PD). METHODS: Six PD patients with hand tremors were studied using a system that employed 3D electromagnetic position sensors to measure the actual, cumulative displacement of the tremoring finger. Patients were studied in different hand positions and activating conditions before and 30, 60, 90 and 120 min after intake of Pramipexole, a dopamine agonist known to reduce tremor. Tremor amplitude and frequency, before and after drug intake, were compared using Mann-Whitney U test and Wilcoxon rank test, respectively. RESULTS: The motion analysis system allowed discrimination of tremor related events from movement artifact and allowed the calculation of real world movement of the finger tremor despite altered hand positions and orientation. Average 3D tremor frequency ranged from 3.71 to 4.34 Hz. Median tremor amplitude (total distance traveled per 5 s interval) decreased with drug from 4.9 to 1.6 cm for resting tremor, 4.5 to 3.7 cm for postural tremor, 3.4 to 3.3 cm for precision tremor, 10.2 to 3.3 cm for tapping activation and 108.6 to 5.7 cm for counting activation. CONCLUSIONS: Our method of 3D analysis provides a robust, single quantitative measure of tremor amplitude that is intuitive and likely to reflect the functional impact of tremor. This methodology should be useful in comparing tremor across patients and in measuring the efficacy of therapeutic interventions. PMID- 10680571 TI - Changes in motor cortex excitability during ipsilateral hand muscle activation in humans. AB - OBJECTIVES: To test whether unilateral hand muscle activation involves changes in ipsilateral primary motor cortex (M1) excitability. METHODS: Single- and paired pulse transcranial magnetic stimulation (TMS) of the right hemisphere was used to evoke motor evoked potentials (MEPs) from the resting left abductor pollicis brevis (APB) in 9 normal volunteers. We monitored changes in motor threshold (MT), MEP recruitment, intracortical inhibition (ICI) and intracortical facilitation (ICF) while the ipsilateral right APB was either at rest or voluntarily activated. Spinal motoneuron excitability was assessed using F-wave recording procedures. RESULTS: Voluntary muscle activation of the ipsilateral APB significantly facilitated the MEPs and F-waves recorded from the contralateral APB. Facilitation was observed with muscle activation >50% of the maximum voluntary force and with stimulus intensities >20% above the individual resting motor threshold. Intracortical inhibition significantly decreased in the ipsilateral M , while there was no significant change in intracortical facilitation during this maneuver. CONCLUSIONS: Unilateral hand muscle activation changes the excitability of homotopic hand muscle representations in both the ipsilateral M1 and the contralateral spinal cord. While the large proportion of MEP facilitation most likely occurred at a spinal level, involvement of the ipsilateral hemisphere may have contributed to the enlargement of magnetic responses. PMID- 10680572 TI - Movement-related potentials in the human spinal cord preceding toe movement. AB - OBJECTIVES: A method by which potentials related to voluntary movement can be recorded noninvasively from the human spinal cord is presented. METHODS: A novel signal processing technique performed on signals recorded by surface electrodes placed over the spinal column was used to filter time-locked back muscle noise, so that the only remaining signals were the spinal movement-related potentials from the brain to the limbs and vice versa. RESULTS: The signals obtained from 7 subjects using this technique are shown and temporally compared with movement related cortical potentials (MRCP) and muscle electromyogram. It is demonstrated that the spinal signal starts approximately 600 ms before the actual movement, and that some features of this signal correspond to changes in cortical potentials. CONCLUSIONS: These findings imply that the spinal cord is not a simple command-carrying medium from the brain to the limbs, and implies that some computational activities take place at the spinal cord level. PMID- 10680573 TI - The short exercise test is normal in proximal myotonic myopathy. AB - OBJECTIVES: Proximal myotonic myopathy (PROMM) is a multisystem disorder that may mimic myotonic dystrophy (MD). Previously we demonstrated that the 60 s exercise test was normal in two siblings with PROMM. The test enabled distinction of PROMM from MD, as there is a well documented immediate post-exercise compound muscle action potential (CMAP) amplitude decline in MD. METHODS: We now performed exercise testing using several exercise durations in 8 PROMM patients from 6 kinships, and one MD patient, extending our previous observations. Repetitive stimulation and needle electromyography findings were also recorded. RESULTS: The 10 (n = 8), 30 (n = 5), and 60 (n = 5) s, and the 5 min (n = 1) exercise tests were normal in all PROMM patients. Specifically, the maximum post-exercise CMAP amplitude decline was 8%. In contrast, the MD patient had CMAP amplitude declines of 48% (10 s exercise test) and 26% (30 s exercise test). The distribution of repetitive stimulation and motor unit duration abnormalities were variable and less diagnostically useful. CONCLUSIONS: The 10, 30, and 60 s exercise tests help distinguish PROMM from MD. As the 10 s exercise test is rapid and easily tolerated, we recommend this test for clinical testing. PMID- 10680574 TI - Asynchronous burst suppression. PMID- 10680575 TI - Biological effects of resveratrol. AB - Resveratrol (3, 4', 5 trihydroxystilbene) is a naturally occuring phytoalexin produced by some spermatophytes, such as grapevines, in response to injury. Given that it is present in grape berry skins but not in flesh, white wine contains very small amounts of resveratrol, compared to red wine. The concentrations in the form of trans- and cis- isomers of aglycone and glucosides are subjected to numerous variables. In red wine, the concentrations of the trans-isomer, which is the major form, generally ranges between 0.1 and 15 mg/L. As phenolic compound, resveratrol contributes to the antioxidant potential of red wine and thereby may play a role in the prevention of human cardiovascular diseases. Resveratrol has been shown to modulate the metabolism of lipids, and to inhibit the oxidation of low-density lipoproteins and the aggregation of platelets. Moreover, as phytoestrogen, resveratrol may provide cardiovascular protection. This compound also possesses anti-inflammatory and anticancer properties. However, the bioavailability and metabolic pathways must be known before drawing any conclusions on the benefits of dietary resveratrol to health. PMID- 10680576 TI - Melatonin synthesis in the greenfrog retina in culture: I. Modulation by the light/dark cycle, forskolin and inhibitors of protein synthesis. AB - Melatonin is synthesized in the pineal gland and the retina of vertebrates. Retinal serotonin N-acetyltransferase (NAT) activity and melatonin show a daily rhythm with high levels during the dark phase of the photocycle. In some vertebrates, these retinal NAT and melatonin rhythms are maintained in vitro. The aim of present work is to develop an eyecup culture system for the greenfrog (Rana perezi), suitable to analyze the mechanisms of regulation of melatonin synthesis by simultaneous determination of NAT activity and melatonin release. The R. perezi eyecups released melatonin to the culture medium in a rhythmic manner at least over a 27-h period under photocycle conditions. NAT activity and melatonin rhythms were similar to that observed in vivo under natural environmental conditions. Rana perezi retina exhibits a pronounced photosensitivity in vitro. Forskolin increased up to 2-fold the NAT activity and 4-fold the melatonin production at any lighting conditions. The addition of the translation inhibitor, cycloheximide, to the medium reduced significantly both nocturnal NAT activity and melatonin release, suggesting that de novo protein synthesis is produced daily during darkness. Actinomycin D, a transcription inhibitor, needs a longer time of action, because pre-existing mRNA must be depleted before the inhibition of melatonin release can be observed. The eyecup culture system is highly sensitive to light and chemical factors, which makes it particularly suitable as a model for the neurochemical analysis of melatonin biosynthesis in the retina of Rana perezi. PMID- 10680577 TI - Melatonin synthesis in the greenfrog retina in culture: II. Dopaminergic and adrenergic control. AB - Serotonin N-acetyltransferase (NAT) activity and melatonin show a daily rhythm with high levels at night. Although the rhythmic properties of NAT and melatonin are similar in pineal gland and retina, great differences in the light perception and transmission mechanisms exist. We have analyzed the effects of adrenergic and dopaminergic agents on greenfrog (Rana perezi) eyecup culture, in order to identify the receptors involved in the regulation of retinal melatonin synthesis. A D2-like receptor is directly involved in the regulation of NAT activity and melatonin release in R. perezi retina. Quinpirole mimics the effect of light, reducing the darkness-stimulated NAT activity and melatonin release, while sulpiride antagonized these actions. Neither D1-agonist (SKF 38393) nor D1 antagonist (SCH 23390) had effect on NAT activity. However, a significant inhibition of darkness-evoked melatonin release was produced by SKF 38393 after 6 hours of culture. The beta- and antagonist1-agonists showed a clear inhibition. However, a direct effect of beta, alpha1 and D1-agonists on photoreceptors is unproven, being more probable that the adrenergic actions imply a non photoreceptor retinal cell. In conclusion, eyecup culture of Rana perezi revealed a dopaminergic control of melatonin synthesis and a possible modulation of dopaminergic tone by adrenergic receptors. Melatonin release is a more sensitive parameter than NAT activity to the action of neuroactive agents, suggesting that melatonin synthesis can be regulated by more than one enzymatic step in Rana perezi. PMID- 10680578 TI - Effect of vitamin E on the response to ischemia-reperfusion of Langendorff heart preparations from hyperthyroid rats. AB - Hyperthyroidism has been reported to decrease heart antioxidant capacity and increase its susceptibility to in vitro oxidative stress. This may affect the heart response to ischemia-reperfusion, a condition that increases free radical production. We compared the functional recovery from in vitro ischemia reperfusion (Langendorff) of hearts from euthyroid (E), hyperthyroid (H, ten daily intraperitoneal injections of T3, 10 microg/100g body weight), vitamin E treated (VE, ten daily intramuscular injections, 20 mg/100g body weight) and hyperthyroid vitamin E-treated (HVE) rats. We also determined lipid peroxidation, tissue antioxidant capacity and the tissue capability to face an oxidative stress in vitro. A significant tachycardia was displayed during reperfusion following 20 min ischemia by the hyperthyroid hearts, together with a low recovery of left ventricular developed pressure (LVDP) and left ventricular dP/dt(max). When H hearts were paced at 300 beats/min, the functional recovery (LVDP and dP/dt(max)) was close to 100% and significantly higher than in E paced hearts. At the end of the ischemia-reperfusion protocol, myocardium antioxidant capacity was significantly lower, whereas lipid peroxidation and the susceptibility to in vitro oxidative stress were higher in the T3 treated (H) than in euthyroid rats. The in vitro tachycardic response, the reduction in the antioxidant capacity and the increase in lipid peroxidation were prevented by treatment of hyperthyroid rats with vitamin E (HVE). These results suggest that the tachycardic response to reperfusion following chronic T3 pretreatment was associated with the reduced capability of the heart to face oxidative stresses in hyperthyroidism. PMID- 10680579 TI - Antioxidative activity of natural products from plants. AB - A variety of flavonoids, lignans, an alkaloid, a bisbenzyl, coumarins and terpenes isolated from Chinese herbs was tested for antioxidant activity as reflected in the ability to inhibit lipid peroxidation in rat brain and kidney homogenates and rat erythrocyte hemolysis. The pro-oxidant activities of the aforementioned compounds were assessed by their effects on bleomycin-induced DNA damage. The flavonoids baicalin and luteolin-7-glucuronide-6'-methyl ester, the lignan 4'-demethyldeoxypodophyllotoxin, the alkaloid tetrahydropalmatine, the bisbenzyl erianin and the coumarin xanthotoxol exhibited potent antioxidative activity in both lipid peroxidation and hemolysis assays. The flavonoid rutin and the terpene tanshinone I manifested potent antioxidative activity in the lipid peroxidation assay but no inhibitory activity in the hemolysis assay. The lignan deoxypodophyllotoxin, the flavonoid naringin and the coumarins columbianetin, bergapten and angelicin slightly inhibited lipid peroxidation in brain and kidney homogenates. It is worth stressing that the compounds with antioxidant effects in this assay, with the exception of tetrahydropalmatin and tanshinone I, have at least one free aromatic hydroxyl group in structure. Obviously, the aromatic hydroxyl group is very important for antioxidative effects of the compounds. None of the compounds tested exerted an obvious pro-oxidant effect. PMID- 10680580 TI - Antioxidative and free radical scavenging activities of selected medicinal herbs. AB - The antioxidative and superoxide- and hydroxyl radical-scavenging activities and pro-oxidant effect of twelve selected medicinal herbs were studied. The aqueous extracts of Coptis chinensis, Paeonia suffruticosa, Prunella vulgaris and Senecio scandens exhibited the highest potency in inhibiting rat erythrocyte hemolysis and lipid peroxidation in rat kidney and brain homogenates. The aforementioned four herbs also demonstrated strong superoxide- and hydroxyl radical-scavenging activity, but exerted only a slight pro-oxidant effect. PMID- 10680581 TI - Lack of effects of glycineB receptor ligands on the psychostimulant-induced discriminative stimuli in rats. AB - To examine the role of glycineB receptors in the stimulus effects induced by psychostimulants, separate groups of rats were trained to discriminate amphetamine (AMPH; 1 mg/kg) from saline (SAL), or cocaine (COC; 10 mg/kg) from SAL, using a two-lever operant procedure. Substitution studies showed that neither 1-aminocyclopropanecarboxylic acid (ACPC; 200 mg/kg) nor 7-chloro-4 hydroxy-3-(3-phenoxy)phenyl-(H)quinolone (L-701,324; 3 mg/kg), being a partial agonist or an antagonist at glycineB receptors, respectively, generalized for the training drugs. Combination tests of glycineB ligands demonstrated that injection of a fixed dose of ACPC (200 mg/kg) or L-701,324 (3 mg/kg) together with different doses of AMPH or COC practically did not modify dose-response curves of the psychostimulants, nor did it affect their ED50 values. Our results indicate that glycineB receptors do not play a role in the discriminative effects of AMPH and COC. PMID- 10680582 TI - Treatment and survival study in the C57BL/6J-APC(Min)/+(Min) mouse with R flurbiprofen. AB - Our previous studies with the mouse model of familial adenomatous polyposis (FAP), C57BL/6J-APC(Min)/+ or Min mouse, demonstrated the optimal dose for adenoma reduction with R-flurbiprofen was 10 mg/kg/day as an undivided dose. Divided doses exhibited no increased efficaciousness. This study examines 10 mg/kg R-flurbiprofen daily (qd) on survival as well as a second daily (q.o.d.) schedule and compares it with sulindac sulfone. The q.o.d. schedule at 10 mg/kg was equally efficacious as qd treatment at the same dose. For the q.o.d. group, tumor number decreased similarly (p<0.01); while body weight gain (p<0.01), hematocrit and average tumor area (both, p<0.05) were improved compared with qd treatment. Treatment with R-flurbiprofen (10 mg/kg/day) increased survival significantly (p=0.0004, log-rank) compared to vehicle treated animals. Major biological endpoints (hematocrit, weight gain, tumor number, average and total area [99% reduction]) were significantly improved in treated animals (p<0.01). Sulindac sulfone treatment (50 mg/kg/day) of the Min mouse produced no significant biological benefit. The dose schedule study suggests that for tumor reduction it is necessary to attain a threshold drug-level but not necessarily sustain it over 24 hrs (pharmacodynamic t1/2 >> pharmacokinetic t1/2). During the period of administration R-flurbiprofen dramatically prolongs survival for the mouse model of the human disease, FAP. PMID- 10680583 TI - The mechanism underlying the hypocholesterolaemic activity of aqueous celery extract, its butanol and aqueous fractions in genetically hypercholesterolaemic RICO rats. AB - Drinking aqueous celery extract for 8 weeks caused a significant reduction in serum total cholesterol (TC) level in growing genetically hypercholesterolaemic (RICO) rats. In addition, administration of butanol fraction (Fbu) and aqueous fraction (Faq) of celery extract for 7 days by intraperitoneal (i.p.) infusion effectively decreased the serum TC and high-density lipoprotein cholesterol (HDL C) levels of adult RICO rats. The 8-week study showed that oral intake of celery extract could enhance the 14C-cholesterol/metabolites excretion. The liver and small intestinal sterol synthesis were not affected. Also, long term drinking of aqueous celery extract did not lead to any undesirable side effects on liver functions. The Fbu and Faq lowered serum TC level mainly through increased bile acid excretion but not by modulating the activity of the rate-limiting enzyme for cholesterol biosynthesis, HMG-CoA reductase. Hence, the mechanism elucidated supports that suggested by the 8-week study. A preliminary chemical characterisation of Fbu and Faq fractions by thin layer chromatography (TLC) showed the presence of sugars and amino acids. There is a possibility that polar compounds with sugar or amino acid side chains(s) could contribute to the hypocholesterolaemic action of celery extract. PMID- 10680584 TI - Genetic evidence that cocaine and caffeine stimulate locomotion in mice via different mechanisms. AB - The effects of cocaine and caffeine on motor activity in two mouse strains 129/OlaHsd (129) and C57BL/6J (C57) were compared. The former mice exhibited lower basal motor activity than the latter. Cocaine (3, 10, 30 mg/kg) injected i.p. in habituated C57 mice produced a dose-dependent increase in rearing, motility and locomotion. In 129 mice, little or no stimulation was seen and only with the highest dose of cocaine. In both strains caffeine (3, 15, 30 mg/kg) produced a dose-dependent increase in rearing, motility and locomotion. The effect of caffeine on rearing was greater in C57 than in 129 mice, but motility and locomotion were stimulated approximately to the same degree in both strains. Thus, differences in the sensitivity to caffeine and cocaine between mouse strains provide genetic evidence that these two stimulants probably produce locomotor stimulation via somewhat different mechanisms. PMID- 10680585 TI - Orphanin FQ/nociceptin inhibits morphine withdrawal. AB - The influence of orphanin FQ/nociceptin (OFQ/N) on the morphine-withdrawal symptom was investigated. Withdrawal syndrome was induced in the morphine dependent rats by an intraperitoneal (i.p.) injection of 2 mg/kg naloxone hydrochloride--an opioid receptors antagonist. Wet-dog shakes were used as a measure of the abstinence syndrome. Intraventricular injections of OFQ/N (5-20 microg/animal) caused significant inhibition of the withdrawal signs at doses between 15-20 microg, in the morphine-dependent rats. OFQ/N alone did not change behavior of the morphine-dependent animals. The obtained results indicate that OFQ/N can inhibit the morphine withdrawal symptoms induced by naloxone. PMID- 10680586 TI - Evaluation of carcinogenic and co-carcinogenic potential Quinalphos in mouse skin. AB - Quinalphos [O,O-diethyl-O-quinoxalinyl-phosphorothidate] is an organophosphorus pesticide with tremendous utility in mixed pest control due to its insecticidal and acaricidal properties. Apart from its pesticidal property, Quinalphos is known to induce various toxic effects in nontarget species and experimental animals. No studies have been conducted to evaluate the carcinogenic/co carcinogenic hazards associated with Quinalphos exposure. In the present set of investigations, the tumorigenic potential of Quinalphos has been evaluated following topical exposure in Swiss albino mice. Long-term animal bioassays conducted for the evaluation of complete carcinogenic, tumour-initiating and tumour-promoting potential of Quinalphos revealed that it has only tumour initiating potential at a dose of 10 mg/kg body weight (b.wt.), in the two-stage mouse skin model of carcinogenesis. Quinalphos exposure failed to produce neoplasia when tested for complete carcinogenic activity at all three tested dose levels or tumour promoting activity. PMID- 10680587 TI - Effect of Withania somnifera on cyclophosphamide-induced urotoxicity. AB - Administration of an extract from the plant Withania somnifera (Family: Solanaceae) (20 mg/dose/animal; i.p.) for five days along with cyclophosphamide (CTX) (1.5 mmol/kg body wt. i.p.) reduced the CTX induced urotoxicity. Morphological analysis of the bladders of the CTX-treated group showed severe inflammation and dark coloration whereas CTX along with the Withania-treated group showed normal bladder morphology. The extract was found to reduce the protein level in the serum (7.92 g/l) after 4 h of CTX treatment, which was higher in the CTX alone-administered group (11.44 g/l). Blood urea N2 level which was drastically enhanced (136.78 mg/100 ml) 2 after the CTX treatment was significantly reduced (52.08 mg/100 ml) when the animals were treated with Withania extract. Similarly the glutathione (GSH) content in both bladder (1.55 micromol/mg protein) and liver (3.76 micromol/mg protein) was enhanced significantly (P<0.001) in the Withania-treated group compared with the CTX alone treated animals (bladder 0.5 micromol/mg protein; liver 1.2 micromol/mg protein) Histopathological analysis of the bladder of CTX alone-treated group showed severe necrotic damage where as the Withania somnifera-treated group showed normal bladder architecture. PMID- 10680588 TI - Inhibitory effect of Coptidis Rhizoma and berberine on the proliferation of human esophageal cancer cell lines. AB - Our previous study demonstrated that the herbal medicine, Oren-to, had antitumor effects on esophageal cancer cells (ECCs) in vitro. The purpose of this study was to examine which of the seven constituents of Oren-to had antitumor effects on esophageal cancer cells. MTT assay showed that, of the seven constituents, only the aqueous extract of Coptidis Rhizoma had potent inhibitory effect on the proliferation of two types of ECC lines, YES-3 and YES-4. In addition, the proliferation of all six types of ECC lines (YES-1 to YES-6) was inhibited in a dose-dependent manner (P<0.001 for all), when co-cultured at each concentration of Coptidis Rhizoma for 72 h. The ID50 of Coptidis Rhizoma for YES-1 to YES-6 was 2.2 microg/ml, 3.0 microg/ml, 0.25 microg/ml, 2.8 microg/ml, 2.5 microg/ml, and 0.5 microg/ml, respectively, berberine, one of protoberberine components of Coptidis Rhizoma, showed potent antitumor effects on all six types of ECC lines as well as Coptidis Rhizoma. In addition, the ID50 of berberine showed a positive correlation with that of Coptidis Rhizoma in six types of ECC lines examined (r2 = 0.763, P = 0.023). Cell cycle analysis of Coptidis Rhizoma-treated cancer cells showed the accumulation of cells in the G0/G1 phase and relative decrease of the S phase. These results support the possibility that the use of Coptidis Rhizoma containing abundant berberine may be useful as one of alternative therapies for esophageal cancers. PMID- 10680589 TI - Proliferation of colonic lymphocytes in response to inflammatory cytokines is lower in mice fed fish oil than in mice fed corn oil. AB - The literature supports an association of chronic inflammation with the development of tumors. The colon contains a specialized lymphocyte population that may influence various stages of colon carcinogenesis. Dietary factors are known to affect both inflammation and tumor development in this tissue. Diets high in n-6 fatty acids are considered to be proinflammatory and tumor-promoting whereas n-3 fatty acids are not. This study examined the proliferative response of colonic lymphocytes (CL) from BALB/c mice fed a diet high in either corn oil or menhaden oil when cultured in the presence of proinflammatory cytokines. CL were isolated from mice fed one of the experimental diets and cultured in the presence of anti-CD2, IL-1beta, IL-2, or TNF-alpha. Proliferation of CL in culture was measured by BrdU incorporation. CL from mice fed the high n-3 diet showed lower rates of proliferation following exposure to the inflammatory cytokines than CL from mice fed the high n-6 diet. CL from the high n-3 group showed the same proliferative potential as those from the n-6 diet group following exposure to a combination of anti-CD2 and TNF. Results from this study indicate that diets high in n-3 fatty acids slow the inflammatory response in the colon as compared to diets high in n-6 fatty acids. The n-3 lipids do not appear to compromise overall immune potential, however. PMID- 10680590 TI - Suppression of azoxymethane-induced colonic aberrant crypt foci by a nitric oxide synthase inhibitor. AB - Nitric oxide synthase (NOS), an important bioregulator of a variety of biological processes, is overexpressed in colonic tumors of humans and rodents. In this study, effects of L-N(G)-nitroarginine methyl ester (L-NAME), a NOS inhibitor, on development of aberrant crypt foci (ACF) induced by azoxymethane (AOM) in F344 male rats were investigated. Six-week-old male F344 rats were fed diets containing 0 or 100 ppm L-NAME, and given s.c. injections of AOM at 15 mg/kg body wt, once a week for 2 weeks. At 17 weeks of age, all animals were sacrificed and their colons were evaluated for numbers of ACF. Feeding of 100 ppm L-NAME inhibited the development of ACF in different sizes by 24-39%, those containing four or more crypts being most markedly affected. Assessment of silver-stained nucleolar organizer regions protein (AgNORs)/nucleus further revealed a 44% reduction by administration of L-NAME. These results suggest that the NOS inhibitor, L-NAME, may be an effective chemopreventive agent against colon carcinogenesis due to depression of cell proliferation. PMID- 10680591 TI - Anti-malignin antibody in serum and other tumor marker determinations in breast cancer. AB - In this study, 154 healthy volunteers and 76 patients were tested using the anti malignin antibody in serum (AMAS) test. Of the 154 volunteers, three were AMAS positive. After further examination, two were positive for cancer and one had a history of ulcerative colitis. Tumor biopsies of 43 suspicious mammography patients revealed that 32 were cancerous and 11 were benign by pathology. For the cancer patients, 31/32 were positive for the AMAS test, while 4/11 of the pathological benign cases were AMAS positive. In comparison to cancer antigen tests, AMAS was more sensitive (97%) in detecting breast cancer than CEA (0%), CA 15-3 (10%), CA 19-5 (5%) or CA 125 (16%) in the same patients. PMID- 10680592 TI - Absence of CD83-positive mature and activated dendritic cells at cancer nodules from patients with hepatocellular carcinoma: relevance to hepatocarcinogenesis. AB - Mature and activated dendritic cells (CD83-positive DCs) are essential for the recruitment and survival of activated tumor-specific lymphocytes during carcinogenesis. The frequencies of CD83 positive DCs were almost same in peripheral blood from patients with hepatocellular carcinoma (HCC) and cirrhosis of liver (LC). However, the numbers of CD83 positive DCs in liver tissues were significant lower in HCC compared with LC (6.6+/-10.9 vs. 33.3+/-24 DCs/specimen, P<0.05). Most importantly, there were no CD83-positive DCs at cancer nodules in HCC. A role of infiltration of activated DCs in liver during hepatocarcinogenesis is shown. PMID- 10680593 TI - Summation of initiation activities of low doses of the non-hepatocarcinogen 1,2 dimethylhydrazine in the liver after carbon tetrachloride administration. AB - Summation of initiation by low doses of the indirect-acting non-hepatocarcinogen, 1,2-dimethylhydrazine (DMH) after proliferative stimulation with a necrogenic dose of carbon tetrachloride (CCl4) was investigated in terms of the induction of glutathione S-transferase placental form (GST-P) positive liver cell foci. Cell kinetics of liver after CCl4 i.g. treatment were examined with bromodeoxyuridine (BrdU) labeling (experiment I). To assess the correlation between cell proliferation and induction of liver cell foci, DMH (10 mg/kg i.g.) was administrated to 7-week-old male F344 rats at 12, 24, 36, 48, 60, 96 h after CCl4 i.g. and initiated populations expanded using the resistant hepatocyte model (experiment IIA). Subsequently, effects of repeated administration (10 mg/kg, four times, i.g.) of DMH were compared with the results of a single administration (40 mg/ kg, i.g.) with the same total dose (experiment IIB). In experiments I and IIA, the numbers and areas of GST-P-positive foci increased with the BrdU labeling index at the time of DMH treatment (maximum after 60 h). In experiment HB, repeated exposure of DMH at 10 mg/kg, four times resulted in significant (P<0.05) increase in number and area of GST-P-positive foci compared with the single administration (40 mg/kg). Thus, multiple low dose treatments during cell proliferation might be most effective for detection of weak initiation activity. PMID- 10680594 TI - Caspase-9 regulates cisplatin-induced apoptosis in human head and neck squamous cell carcinoma cells. AB - The aim of this study was to understand the molecular requirements for cisplatin induced apoptosis in human head and neck squamous cell carcinoma (HNSCC) cell death. Cisplatin induced apoptosis in HNSCC cell lines, HSC-2, HSC-3 and HSC-4 in a dose-dependent manner. However, cisplatin did not induce the expression of Fas ligand mRNA or upregulation of Fas protein. By caspase activation assays, cisplatin induced Caspase-3 (Casp-3), -8 and -9 activation. In all three lines tested, the use of a specific inhibitor of Casp-9 almost completely blocked cisplatin-induced apoptosis, while the use of Casp-3 and -8 inhibitors resulted in a partial blockade of cisplatin-induced apoptosis. Our results strongly suggest that Casp-9-dependent apoptosis plays an important role in cisplatin induced HNSCC apoptosis. PMID- 10680595 TI - Aberrant DNA methylation precedes loss of heterozygosity on chromosome 16 in chronic hepatitis and liver cirrhosis. AB - The aim of this study was to examine the significance of aberrant DNA methylation, the participation of which in genetic instability is controversial, in hepatocarcinogenesis. The DNA methylation status of the region around the promoter of the E-cadherin tumor suppressor gene, which is located on 16q22.1, and the allelic status at the D16S421 locus, which is adjacent to the E-cadherin locus, were examined using microdissected liver specimens from 38 hepatocellular carcinoma (HCC) patients. Almost all of the non-cancerous liver tissues showed histological findings compatible with chronic hepatitis and cirrhosis, which are considered to be precancerous conditions. DNA hypermethylation was detected in 61% of the non-cancerous liver tissues. The incidence of DNA hypermethylation in the non-cancerous liver tissues of patients with HCCs also showing DNA hypermethylation (72%) was significantly higher than that of patients without DNA hypermethylation in their HCCs (53%, P<0.05). Loss of heterozygosity (LOH) at the D16S421 locus was detected in 35% of the non-cancerous liver tissues. The incidence of LOH in the non-cancerous liver tissues of patients with HCCs also showing LOH was 78%, whereas LOH was not detected in non-cancerous liver tissues of patients without LOH in their HCCs. Fifty-two percent of the non-cancerous liver tissues showed both or neither of DNA hypermethylation and LOH; the incidence of DNA hypermethylation alone in noncancerous liver tissue was 41%. The incidence of LOH alone in non-cancerous liver tissue (7%) was significantly lower compared to those of the former two cases (P<0.0001). These data suggest that aberrant DNA methylation participates in the precancerous stage of hepatocarcinogenesis by preceding, or causing, LOH. PMID- 10680596 TI - Trans-4-hydroxy-2-nonenal, an aldehydic lipid peroxidation product, lacks genotoxicity in lacI transgenic mice. AB - In order to cast light on the significance of lipid peroxidation products for carcinogenesis, the lacI mutant frequency (MF), micronucleus induction and cell proliferation were analyzed in lacI transgenic mice treated with trans-4-hydroxy 2-nonenal (HNE), a typical example. Male mice were ip injected with HNE at doses of 0, 5 or 50 mg/kg bw and 48 h thereafter, peripheral blood was collected for analyzing micronucleus induction. After 14 days, the mice were sacrificed to allow tissue sampling for examination of lacI MF and cell proliferative activity. Sixty percent of the mice given 50 mg/kg HNE died within 5 days after the treatment, but no other mortalities were observed. Histopathologically, marked pulmonary hemorrhage was found in the 50 mg/kg HNE group mice that survived until day 14. Immunohistochemically, HNE-modified proteins were detected in their alveolar macrophages. The HNE treatment did not increase lacI MF in the liver, kidney and lung and no significant increase in micronucleus induction or cell proliferation in major organs was found in either treatment. Moreover, no tumors developed in the 5 mg/kg HNE-treated mice which survived until week 78. Our results thus indicate that HNE lacks in vivo genotoxicity in lacI transgenic mice even when lethal doses are applied. PMID- 10680597 TI - Differential expression of an estrogen receptor messenger RNA containing exon 1' sequences in MCF-7 breast cancer cell line stocks. AB - The presence of an exon 1' sequence in the estrogen receptor alpha (ERalpha) mRNA was detected in different stocks of ER-positive MCF-7 human breast cancer cells by reverse transcriptase polymerase chain reaction (RT-PCR) and ribonuclease protection analysis (RPA), but not by Northern blot analysis. This mRNA, however, was not detectable in ERalpha-positive ZR-75-1 or ERalpha-negative MDA-MB-231 breast cancer cells, suggesting that exon 1' ER mRNA is differentially expressed in some but not all ER-positive cell lines, and then, only at very low levels. PMID- 10680598 TI - Analysis of highly frequent allelic loss region on distal chromosome 12 in murine radiation-induced lymphomas. AB - Recent genetic studies of tumorigenesis have strongly suggested an existence of tumor suppressor gene(s) on murine chromosome 12 and human chromosome 14q32. We previously described that putative tumor suppressor gene(s) might reside between D12Mit53 and D12Mit233. We analyzed three genes, Tcl1, Yy1 and Tnfalphaip2, which had been mapped around the region, as the candidates in radiation lymphomagenesis of (BALB/c x MSM/Ms)F1 hybrid mice. The locus order and distances of the three genes and microsatellite loci were estimated as follows: [centromere] - Tcl1-(> or =0.085 cM)-D12Mit50-(0.085 cM)D12Mit132-(1.96 cM)D12Mit122-(0.085 cM)D12Mit53 (1.37 cM)-[D12Mit233,D12Mit279,Yy1]-(0.085 cM)-D12Mit181-(> or =0.17 cM) Tnfalphaip2 - [telomere]. Allele losses at Tcl1, Yy1 genes and D12Mit233 were observed in 94(45%), 143(68%) and 147(70%) of 210 lymphomas, respectively. In semi-quantitative analysis of Yy1 mRNA levels by RT-PCR, kinetics of the yield of the Yy1-cDNA-specific PCR products showed almost the same profiles among thymic lymphomas with allelic loss at Yy1, lymphomas with both alleles retained and normal thymus. These results suggest that Tcl1, Yy1 and Tnfalphaip2 genes are not predominantly involved in radiation lymphomagenesis of mice. In further analysis of the common allelic loss region, we found new loci, Y152pR1 and Y184pR2, from YACs which located in the hot region between D12Mit53 and D12Mit233, and the highest frequency of allelic loss (71%) was observed at the Y184pR2 locus. The LOH patterns of individual lymphomas suggest that putative tumor suppressor gene(s) lies between Y152pR1 and Y184pR2. PMID- 10680599 TI - Involvement of retinoblastoma protein in p27Kip1-induced apoptosis. AB - p27Kip1, a cyclin-dependent kinase (CDK) inhibitor, plays a critical role in cell cycle regulation. Expression of p27Kip1 is shown to increase during apoptosis in mammalian cells. Here, to directly address the role of p27Kip1 in apoptosis, p27Kip1 is overexpressed in human SK-Hep1 hepatoma cells. This leads to apoptotic cell death and this reduces protein, but not mRNA, levels of the retinoblastoma (Rb). Consistently, accumulation of Rb protein blocks p27Kip1-mediated apoptosis. These studies demonstrate an involvement of Rb in the apoptotic cell death which is induced by overexpression of p27Kip1. PMID- 10680600 TI - Guidelines for surveillance, prevention, and control of West Nile virus infection -United States. AB - The introduction of West Nile (WN) virus in the northeastern United States during the summer and fall of 1999 raised the issue of preparedness of public health agencies to handle sporadic and outbreak-associated vector-borne diseases. In many local and state health departments, vector-borne disease capacity has diminished. Because it is unknown whether the virus can persist over the winter, whether it has already or will spread to new geographic locations, and the public health and animal health implications of this introduction, it is important to establish proactive laboratory-based surveillance and prevention and control programs to limit the impact of the virus in the United States. On November 8 and 9, 1999, CDC and the U.S. Department of Agriculture (USDA) cosponsored a meeting of experts representing a wide range of disciplines to review the outbreak and to provide input and guidance on the programs that should be developed to monitor WN virus activity and to prevent future outbreaks of disease. This report summarizes the guidelines established during this meeting. PMID- 10680601 TI - Accutane-exposed pregnancies--California, 1999. AB - Accutane (Roche Laboratories, Nutley, New Jersey), known by the generic name "isotretinoin," is a prescription oral medication approved by the Food and Drug Administration (FDA) to treat severe, recalcitrant nodular acne. It is also a known human teratogen that can cause multiple major malformations. Embryopathy associated with the mother's exposure to isotretinoin during the first trimester of pregnancy includes craniofacial, cardiac, thymic, and central nervous system malformations . In response to FDA recommendations, the manufacturer began a pregnancy-prevention program (PPP) in 1988 that included educational materials for physicians and patients and offered women reimbursement for contraceptive counseling by a physician. The PPP coordinators asked reproductive-aged women being treated with isotretinoin to enroll voluntarily in the Boston University Accutane Survey (BUAS). The total number of reproductive-aged women taking isotretinoin in the United States is unknown; however, 454,273 women enrolled in the BUAS from 1989 to October 1999. BUAS has estimated that 38%-40% of reproductive-aged women taking isotretinoin chose to enroll in the survey (BUAS, unpublished data, 1999). Although isotretinoin is contraindicated in pregnancy and has a package label warning users to avoid pregnancy while taking it, exposed pregnancies occur. Approximately 900 pregnancies occurred among BUAS enrollees during 1989-1998 (BUAS, unpublished data, 1999). Roche Laboratories began direct to-consumer print advertisements in 1996, added television and radio advertisements to selected cities in 1997, and expanded the campaign to the entire United States in 1998. PMID- 10680602 TI - Update: raccoon rabies epizootic--United States and Canada, 1999. AB - In 1977, an outbreak of raccoon rabies was detected in an area on the West Virginia-Virginia border. Since then, the area affected by this distinct variant of rabies virus associated with raccoons has spread to Ohio in the west and New York, Pennsylvania, Vermont, New Hampshire, and Maine in the north. In addition, the once separate epizootics of rabies among raccoons in the southeastern and mid Atlantic states converged in North Carolina. In July 1999, the raccoon rabies virus variant was reported from Ontario, Canada, on the New York border. This report describes the spread of this epizootic of raccoon rabies through mid Atlantic and northeastern states and into Canada. PMID- 10680603 TI - Recommended childhood immunization schedule--United States, 2000. AB - Each year, CDC's Advisory Committee on Immunization Practices (ACIP) reviews the recommended childhood immunization schedule to ensure it remains current with changes in manufacturers' vaccine formulations, revisions in recommendations for the use of licensed vaccines, and recommendations for newly licensed vaccines. This report presents the recommended childhood immunization schedule for 2000 and explains the changes that have occurred since January 1999. PMID- 10680604 TI - HPMA copolymer-modified avidin: immune response. AB - Protein-polymer conjugates to be used in the pretargeted delivery of a photosensitizer to cells were synthesized and characterized. Avidin was modified by N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers bearing the photosensitizer, mesochlorin e6 mono(N-2-aminoethylamide) (Mce6). Synthesis of HPMA copolymer-avidin-Mce6 conjugates was carried out so that either predominantly single point attachment or multipoint attachment of copolymer chains to avidin would result. HPMA copolymer-avidin conjugates were used which retained specific binding activity to a lower affinity biotin analog. Antigen specific anti-avidin immune response was shown to be reduced six-fold in some HPMA copolymer-avidin conjugates when compared to immune response to unmodified avidin. HPMA copolymer itself was shown to elicit a very low (IgM) immune response. PMID- 10680605 TI - Protein A immobilization and HIgG adsorption onto porous/nonporous and swellable HEMA-incorporated polyEGDMA microspheres. AB - Both non swellable and swellable poly(EGDMA/HEMA) microbeads were produced by suspension copolymerization. These microbeads were modified by immobilization of a spacer-arm (hexamethylene diamine (HMDA)) and protein A. The optimal values for modifications were as follows: sodium periodate concentration, 1.0 mgml(-1); HMDA concentration, 4 mgml(-1); and glutaraldehyde concentration, 0.070 microgml(-1). Adsorption of protein A onto the plain and periodate oxidized poly(EGDMA/HEMA) microbeads were very close to each other, and were 0.01-0.02 mg protein A on the 1-g Microbeads I and II, respectively. Protein A immobilization on poly(EGDMA/HEMA) microbeads were studied at different temperatures, times, and pHs using single protein solution containing different amounts of proteins. The optimal values for immobilization were as follows: the initial protein A concentration, 0.1 mgml(-1); temperature, 25 degrees C; pH, 9.5; and immobilization time, 120 min. Incorporation of protein A resulted in 1.420 and 1.825 mg protein A on the 1-g Microbeads I and II, respectively. HIgG adsorption capacity on the protein A-incorporated poly(EGDMA/HEMA) microbeads is 27 and 35 mg HIgGg(-1) polymer for Microbeads I and II, respectively. PMID- 10680606 TI - Controlled release of antihypertensive drug from the interpenetrating network poly(vinyl alcohol)-guar gum hydrogel microspheres. AB - Poly(vinyl alcohol)-guar gum interpenetrating network microspheres were prepared by cross-linking with glutaraldehyde. Nifedipine, an antihypertensive drug, was loaded into these matrices before and after cross-linking to study its release patterns. The extent of cross-linking was analyzed by Fourier transform infrared spectroscopy and differential scanning calorimetry. Furthermore, the microspheres were characterized for drug entrapment efficiency, particle size, transport of water into the matrix and drug release kinetics. Scanning electron microscopic photographs confirmed the spherical nature and surface morphology. The mean particle size of the microspheres was found to be around 300 microm. The molecular transport phenomenon, as studied by the dynamic swelling experiments, indicated that an increase in cross-linking affected the transport mechanism from Fickian to non-Fickian. The in vitro release study indicated that the release from these microspheres is not only dependent upon the extent of cross-linking, but also on the amount of the drug loaded as well as the method of drug loading. PMID- 10680607 TI - Effect of ionic strength on the loading efficiency of model polypeptide/protein drugs in pH-/temperature-sensitive polymers. AB - In this report, the effect of ionic strength on the loading efficiency of three model polypeptide/protein drugs, namely angiotensin II, insulin, and cytochrome c, in pH- and temperature-sensitive terpolymers of poly(NIPAAm-co butylmethacrylate-co-acrylic acid) (poly(NIPAAm-co-BMA-co-AA)) has been investigated. Loading efficiency of polypeptides in pH-/temperature-sensitive beads composed of poly(NIPAAm-co-BMA-co-AA) terpolymer is predominantly governed by hydrophobic interactions, both nonspecific surface interactions and/or specific interactions (binding pockets) between the protein and the polymer molecules. Thus, loading efficiency increases with ionic strength. However, as ionic strength increases further, polymer deswelling (collapse), which is also controlled by hydrophobic forces, becomes more pronounced, and consequently, a higher fraction of water is squeezed out during bead formation and the loading efficiency starts to decrease. PMID- 10680608 TI - Immunogenicity of antigens in boiled alginate microspheres. AB - Vaccine efficacy can be enhanced by delivery of antigens in synthetic microspheres. The process of antigen incorporation into microspheres can expose fragile antigens to damaging conditions, such as high temperatures, and to bacterial contamination. Maintenance of immunogenicity of several antigens and reduction of bacterial load in alginate microspheres following boiling was evaluated. Mice were immunized subcutaneously, initially and again 21 days later, with either non-boiled or boiled microspheres containing ovalbumin (OVA), a culture supernatant vaccine of Pasteurella haemolytica (PHV), or a potassium thiocyanate extract of P. multocida (PTE). Serum samples were obtained prior to immunization and at the time of euthanasia 28 days later. Culture of microspheres showed that boiling completely eliminated aerobic bacterial growth for OVA containing microspheres, and reduced growth by a factor of 10(4) for PTE microspheres. More bacteria were cultured after boiling than before for PHV microspheres. ELISA performed on serum and intestinal lamina propria explant supernatants showed that immunogenicity of PHV microspheres was not altered by boiling. Boiled OVA microspheres were still able to stimulate a significant serum IgG anti-OVA titer in mice, but boiled PTE microspheres completely lacked immunogenicity. Elispot assays of spleens showed that only PHV microspheres were able to retain immunogenicity after boiling. Results indicate that boiling is not an effective means for reducing the bacterial load of alginate microspheres and that the process is associated with a diminution of vaccine immunogenicity. PMID- 10680609 TI - Modification of the adhesive properties of collagen by covalent grafting with RGD peptides. AB - Collagen, either alone or in combination with other materials, is an important natural biomaterial that is used in a variety of tissue-engineering applications. Cell adhesion and migration of cells within collagen-based biomaterials may be controlled by modifying the adhesive properties of collagen. Furthermore, spatially controlling the adhesiveness of the collagen may allow controlled localization or redistribution of cells. A method is presented for covalently coupling peptides that contain the well-characterized arginine-glycine-aspartic acid adhesion sequence directly to type I collagen monomers prior to fibrillogenesis. A heterobifunctional coupling agent was used to create stable amide and disulfide bonds with the lysine residues of the collagen monomers and the cysteine termini of the peptide molecules, respectively. The degree of conjugation could be controlled by changing the reaction conditions (ratios of reactants added and the length of incubation). The microstructure and gelation times of gels composed of covalently modified collagen were similar to those of unmodified gels. Cell adhesion on adsorbed monolayers of modified collagen was quantified using a well-established clonal cell line (K1735 murine melanoma). Cell adhesion was found to increase with both increasing degree of conjugation and increasing ratio of modified to unmodified collagen. PMID- 10680610 TI - Polymerization and surface analysis of electrically-conductive polypyrrole on surface-activated polyester fabrics for biomedical applications. AB - A new synthetic route is reported for the synthesis and covalent bonding of electrically conductive polypyrrole to a poly(ethylene terephthalate) fabric. It involves a three-step process including surface phosphorylation and graft polymerization from the gaseous phase. In the first step, the fibre surfaces are activated using phosphorus trichloride. Then, 1-(3-hydroxypropyl) pyrrole is introduced and grafted to the phosphorus chloride to create an ester bond between the fibres and the pyrrole. Finally, the pyrrole-grafted fibres are dipped in an aqueous FeCl3 catalyst and exposed to pyrrole monomer vapor for the final polymerization. This last step creates an electrically conductive polypyrrole layer covalently linked to the poly(ethylene terephthalate) fibres. ESCA analysis indicates a high degree of phosphorylation and grafting of the anchor molecules. Scanning electron microscopy reveals an overall smooth and uniform surface coating of polypyrrole on the polyester fibres. The use of ATR-FTIR spectroscopy is not able to distinguish between polypyrrole-coated and non-coated fabrics because of the extremely thin polypyrrole layer. Measurements of dynamic surface wetting indicated that the polypyrrole-coated fabric is more hydrophilic than the untreated control. With values for surface resistivity in the range 10(4)-10(5) ohmz/square, such polypyrrole-coated fabrics are considered attractive candidates for biomedical applications. PMID- 10680611 TI - Novel bifunctional polymer with reactivity and temperature sensitivity. AB - To introduce reactive groups into temperature-responsive polymeric chains of poly(N-isopropylacrylamide) (PIPAAm), IPAAm is copolymerized with other monomer such as acrylic acid (AAc). IPAAm homopolymer exhibited temperature-responsive properties and phase transition at 32 degrees C, however, the lower critical solution temperature (LCST) of the IPAAm-AAc copolymer shifts to a higher temperature and the phase transition becomes insensitive with increasing AAc content. To achieve a useful bifunctional copolymer containing both reactivity and temperature-sensitivity, we assumed that the homopolymer-like structure in the polymer chain would be required to maintain a sensitive temperature response with functional groups. Therefore, we designed a reactive monomer, 2 carboxyisopropylacrylamide (CIPAAm), and investigated its copolymerization with IPAAm. The important characteristic of the poly(IPAAm-co-CIPAAm) structure is that it was composed of the same polymer backbone and isopropylamide groups and some additional carboxyl groups. The transmittance measurement of the polymer aqueous solution revealed that phase transition of IPAAm-co-CIPAAm random copolymer occurred within a very narrow temperature range in pH 6.4, 7.4, and also even 9.0 phosphate buffered solution. These profiles were almost same as that of IPAAm homopolymer. While, under the same conditions, phase transition properties of poly(IPAAm-co-AAc)s solution were considerably influenced by small AAc content. We succeeded with the preparation of bifunctional polymer that possessed reactive functional groups and very sensitive response to temperature change. PMID- 10680612 TI - In situ atomic force microscopic observation of albumin adsorption onto phase separated organosilane monolayer surface. AB - A mixed (n-octadecyltrichlorosilane (OTS)/[2 (perfluorooctyl)ethyl]trichlorosilane (FOETS)) monolayer was prepared on the water subphase and was subsequently immobilized onto the silicon wafer surface by chemical bonds. Atomic force microscopic (AFM) observation of the mixed (OTS/FOETS) monolayer revealed the formation of a phase-separated structure. In situ AFM observation of the adsorption behavior of bovine serum albumin (BSA) onto the mixed (OTS/FOETS) monolayers, successfully showed the adsorption behavior of BSA onto the phase-separated surface. It also revealed that in the case of pH 7.5, BSA was preferentially adsorbed onto the lower surface free energy FOETS phase of the mixed (OTS/FOETS) monolayer. On the other hand, BSA was adsorbed homogeneously onto the OTS and FOETS phases at the isoelectric point of BSA (pI 4.7). These results indicate that the preferential adsorption of BSA onto the FOETS phase in the mixed (OTS/FOETS) monolayer system may be due to: (1) the minimization of interfacial free energy between a monolayer surface and an aqueous solution; and (2) the electrostatic repulsion among BSA molecules bearing negative charges. PMID- 10680613 TI - Mechanical bioeffects from diagnostic ultrasound: AIUM consensus statements. American Institute of Ultrasound in Medicine. PMID- 10680614 TI - Section 2--definitions and description of nonthermal mechanisms. American Institute of Ultrasound in Medicine. PMID- 10680615 TI - Section 3--selected biological properties of tissues: potential determinants of susceptibility to ultrasound-induced bioeffects. American Institute of Ultrasound in Medicine. AB - Although harmful side effects have not been reported in humans associated with the clinical application of continuous wave (CW) ultrasound (typical of therapeutic applications) or pulsed ultrasound (typical of diagnostic applications), bioeffects have been reported in nonhuman mammalian species with both waveforms. Bioeffects have been reported in organ systems that have tissues associated with well-defined gas bodies, such as lung and intestine, and in tissues not associated with well-defined gas bodies, such as nervous tissue, liver, kidney, and reproductive tissues. Lesions in tissues with well-defined gas bodies include hemorrhage in lung and intestine; lesions in tissues without well defined gas bodies include cell and tissue destruction and necrosis with tissue specific hemorrhage in nervous tissue, liver, kidney, and reproductive tissues. Studies suggest that there are unique differences in the responses of tissues to CW and pulsed ultrasound among animal species and within a species based on age or stage of development. Data from reference materials covering the anatomy and histology of animal tissues indicate that important structural differences exist among species and within a species based on age or stage of development. These structural differences and how they influence the responses of tissues and organs to ultrasound appear to be focused on the serosal surfaces (protective coverings) of these organs. Such surfaces include, but are not limited to, the visceral pleura of the lung and the visceral peritoneum of all abdominal organs. The thickness and composition (e.g., structural fiber type, collagen, or elastic fibers) of these surfaces and their contiguous internal structural units, such as septa and trabecula, also appear to be important in determining the susceptibility of a tissue or organ to ultrasound-induced damage. Although data exist that support the hypothesis that there are species differences in susceptibility to CW ultrasound-induced lung hemorrhage, data from studies using pulsed ultrasound are less abundant but suggest that species differences in susceptibility to pulsed ultrasound-induced lung hemorrhage may also possibly exist. Species differences in susceptibility to ultrasound-induced lung hemorrhage appear inversely related to the thickness of the visceral pleura of the lung. Although pulsed ultrasound can induce hemorrhage in the intestine of mice, there are no data from intestinal studies currently available suggesting there are species differences in susceptibility to pulsed ultrasound. There exists, for the intestine, information similar to that reported in the lung, which documents innate tissue and organ differences among mammalian species that could influence susceptibility to pulsed ultrasound. The analysis, interpretation, and speculation regarding bioeffects and tissue properties documented in this section and the conclusions presented likely will mature and evolve as new information becomes available through additional in vivo bioeffects research. PMID- 10680617 TI - Section 5--nonthermal bioeffects in the absence of well-defined gas bodies. AB - The purpose of this section is to describe bioeffects that occur in those instances in which gas bodies are not known to commonly be present. This is in contrast to situations such as lung and intestine, where bioeffects have been proposed to be associated with the known presence of gas (Section 4). In addition, this section does not include those studies in which microbubbles (such as ultrasound contrast agents) are purposefully introduced (Section 6). Therefore, this section includes a large collection of bioeffects that may be related to cavitation activity or may not be related at all to the presence of gas bodies. The intention is to provide an overall summary of the information currently available that pertains to those situations in which gas bodies are not known to be present and to provide this information in support of conclusions and recommendations on this subject. PMID- 10680616 TI - Section 4--bioeffects in tissues with gas bodies. American Institute of Ultrasound in Medicine. AB - Several animal models have exhibited thresholds for petechial hemorrhage in lung within the current output of diagnostic ultrasound systems. In addition, thresholds for damage in the mouse intestine due to diagnostic pulses of ultrasound have been explored. The implications for human lung and intestinal exposure to clinical diagnostic ultrasound have not yet been determined. In this section, the data supporting the thresholds of petechial hemorrhage in these organ systems and the morphological observations will be reviewed. The potential mechanical mechanisms of damage to these organs due to diagnostic ultrasound also will be reviewed. Special attention will be given to the occurrence of inertial cavitation both in vitro and in vivo. The effects of ultrasound parameters, age, and species on the threshold for damage in animal models will be explored. PMID- 10680621 TI - Visual signal detection in structured backgrounds. III. Calculation of figures of merit for model observers in statistically nonstationary backgrounds. AB - Models of human visual detection have been successfully used in computer generated noise. For these backgrounds, which are generally statistically stationary, model performance can be readily calculated by computing the index of detectability d' from the noise power spectrum, the signal profile, and the model template. However, model observers are ultimately needed in more real backgrounds, which may be statistically non-stationary. We investigated different methods to calculate figures of merit for model observers in real backgrounds based on different assumptions about image stationarity. We computed performance of the nonpre-whitening matched-filter observer with an eye filter on mammography and coronary angiography for an additive or a multiplicative signal. Performance was measured either by applying the model template to the images or by computing closed-form expressions with various assumptions about image stationarity. Results show first that the structured backgrounds investigated cannot be considered stationary. Second, traditional closed-form expressions of detectability calculated from the noise power spectra with the assumption of background stationarity lead to erroneous estimates of model performance. Third, the most accurate way of measuring model performances is by directly applying the model template on the images or by computing a closed-form expression that does not assume image stationarity. PMID- 10680622 TI - Visual signal detection in structured backgrounds. IV. Figures of merit for model performance in multiple-alternative forced-choice detection tasks with correlated responses. AB - Many investigators are currently developing models to predict human performance in detecting a signal embedded in complex backgrounds. A common figure of merit for model performance is d', an index of detectability that can be mathematically related to the proportion correct (Pc) when the responses of the model are Gaussian distributed and statistically independent. However, in many multiple alternative forced-choice (MAFC) detection tasks, the target appears in one of M different locations within an image. If the image contains slow spatially varying luminance changes (low-pass noise), the pixel luminance values at the possible signal locations are correlated and therefore the model/human responses to the different locations might also be correlated. We investigate the effect of response correlations on model performance and compare different figures of merit for these conditions. Our results show that use of the standard d' index of detectability assuming statistical independence can lead to erroneous underestimates of Pc and misleading comparisons of models. We introduce a novel figure of merit d'(r) that takes into account response correlations and can be used to accurately estimate Pc. Furthermore, we show that d'(r) can be readily related to the standard index of detectability d' by d'(r) = d'/square root of (1 - r), where r is the correlation between the responses in any MAFC detection task. We illustrate the use of the theory by computing figures of merit for two linear models detecting a signal in one of four locations within medical image backgrounds. PMID- 10680623 TI - Color signals in natural scenes: characteristics of reflectance spectra and effects of natural illuminants. AB - Multispectral images of natural scenes were collected from both forests and coral reefs to represent typical, complex scenes that might be viewed by modern animals. Both reflectance spectra and modeled visual color signals in these scenes were decorrelated spectrally by principal-component analysis. Nearly 98% of the variance of reflectance spectra and color signals can be described by the first three principal components for both forest and coral reef scenes, which implies that three well-designed visual channels can recover almost all of the spectral information of natural scenes. A variety of natural illuminants affects color signals of forest scenes only slightly, but the variation in ambient irradiance spectra that is due to the absorption of light by water has dramatic influences on the spectral characteristics of coral reef scenes. PMID- 10680618 TI - Section 6--mechanical bioeffects in the presence of gas-carrier ultrasound contrast agents. American Institute of Ultrasound in Medicine. AB - This review addresses the issue of mechanical ultrasound-induced bioeffects in the presence of gas carrier contrast agents (GCAs). Here, the term "contrast agent" refers to those agents that provide ultrasound contrast by being composed of microbubbles, encapsulated or not, containing one or more gases. Provided in this section are summaries on how contrast agents work, some of their current uses, and the potential for bioeffects associated with their presence in an ultrasonic field. PMID- 10680624 TI - Relational color constancy in achromatic and isoluminant images. AB - Relational color constancy, which refers to the constancy of perceived relations between surface colors under changes in illuminant, may be based on the computation of spatial ratios of cone excitations. As this activity need occur only within rather than between cone pathways, relational color constancy might be assumed to be based on relative luminance processing. This hypothesis was tested in a psychophysical experiment in which observers viewed simulated images of Mondrian patterns undergoing colorimetric changes that could be attributed either to an illuminant change or to a nonilluminant change; the images were isoluminant, achromatic, or unmodified. Observers reliably discriminated the two types of changes in all three conditions, implying that relational color constancy is not based on luminance cues alone. A computer simulation showed that in these isoluminant and achromatic images spatial ratios of cone excitations and of combinations of cone excitations were almost invariant under illuminant changes and that discrimination performance could be predicted from deviations in these ratios. PMID- 10680625 TI - Chromatic contrast sensitivity: the role of absolute threshold and gain constant in differences between the fovea and the periphery. AB - A model of foveal achromatic and chromatic sensitivity [Vision Res. 36, 1597 (1996)] was extended to the peripheral visual field. Threshold-versus-illuminance functions were analyzed to determine effects of eccentricity on absolute thresholds and gain constants of chromatic and luminance mechanisms. The resulting peripheral model successfully predicted peripheral contrast sensitivity as a function of wavelength, for both white and 500-nm backgrounds. We conclude that the short-wavelength-sensitive cone opponent mechanism may mediate thresholds in Sloan's notch in the normal periphery and that interpretation of reduced chromatic sensitivity in the periphery requires an explicit model of how eccentricity affects both the gain constant and the absolute threshold. PMID- 10680626 TI - Role of perceptual organization in chromatic induction. AB - Color matches between two small patches were made in a display containing ten larger regions of different chromaticities. The spatial organization of the ten regions was varied while keeping constant the immediate surround of each patch as well as the space-average chromaticity of the entire stimulus. Different spatial arrangements were designed to alter the perceptual organization inferred by the observer without changing the ensemble of chromaticities actually in view. For example, one arrangement of the ten regions was consistent with five surfaces under two distinct illuminations, with one edge within the display (an "apparent illumination edge") dividing the stimulus into two areas, one under illuminant A and the other under illuminant C. Another spatial arrangement had the ten regions configured to induce an observer to infer ten surfaces under a single illumination. When the ten regions were arranged with an apparent illumination edge, the patch within the area of illuminant C was perceived as bluer than when the same patch and immediate surround were presented without an apparent illumination edge. The results are accounted for by positing that observers group together regions sharing the same inferred illumination, with a consequent effect on color perception: A fixed patch-within-surround shifts in hue and saturation toward the perceived illumination. We suggest that the change in color perception in a complex scene that results from a difference in real illumination may be caused by the inferred illumination at the perceptual level, not directly by the physical change in the light absorbed by photoreceptors. PMID- 10680619 TI - Section 7--discussion of the mechanical index and other exposure parameters. American Institute of Ultrasound in Medicine. AB - There have been long-term efforts to identify a threshold pressure for the onset of inertial cavitation under conditions relevant to ultrasound in medicine. Before the introduction of the output display standard [AIUM/NEMA, 1992a], quantities such as the spatial peak pulse average intensity (I(SPPA)), and, earlier, Im, the spatial peak intensity averaged over the largest half-cycle, were used to give a measure of the potential of a cavitation-based bioeffect due to an acoustic field. Relatively early in the Output Display Standard development effort, the Food and Drug Administration indicated a need for a superior indicator for the potential for cavitation-related bioeffects, initiating a search for such an index. The following paragraphs give an outline of the steps used to develop the Mechanical Index, its relevance as a potential bioeffects indicator, and some information on other exposure parameters involved in bioeffects research. PMID- 10680627 TI - Invariant cone-excitation ratios may predict transparency. AB - Cone-excitation ratios for pairs of surfaces are almost invariant under changes in illumination and offer a possible basis for color constancy [Proc. R. Soc. London Ser. B 257, 115 (1994)]. We extend this idea to the perception of transparency on the basis of the close analogy between the changes in color signals that occur for surfaces when the illumination changes and the changes in color signals when the surfaces are covered by a filter. This study presents measurements and simulations to investigate the conditions under which cone excitation ratios are statistically invariant for physically transparent systems. The invariance breaks down when the spectral transmission of the filters is low at some or all wavelengths. We suggest that cone-excitation ratios might be useful to define the stimulus conditions necessary for the perception of transparency. PMID- 10680628 TI - Image restoration with the Viterbi algorithm. AB - The Viterbi algorithm (VA) is known to given an optimal solution to the problem of estimating one-dimensional sequences of discrete-valued pixels corrupted by finite-support blur and memoryless noise. A row-by-row estimation along with decision feedback and vector quantization is used to reduce the computational complexity of the VA and allow the estimation of two-dimensional images. This reduced-complexity VA (RCVA) is shown to produce near-optimal estimation of random binary images. In addition, simulated restorations of gray-scale images show the RCVA estimates to be an improvement over the estimates obtained by the conventional Wiener filter (WF). Unlike the WF, the RCVA is capable of superresolution and is adaptable for use in restoring data from signal-dependent Poisson noise corruption. Experimental restorations of random binary data gathered from an optical imaging system support the simulations and show that the RCVA estimate has fewer than one third of the errors of the WF estimate. PMID- 10680629 TI - Polarization and statistical analysis of scenes containing a semireflector. AB - We present an approach to recover scenes deteriorated by reflections off a semireflecting medium (e.g., a glass window). The method, based on imaging through a polarizer at two or more orientations, separates the reflected and transmitted scenes and determines which is which. We analyze the polarization effects, taking into account internal reflections within the medium. The scene reconstruction requires the estimation of the orientation (inclination and tilt angles) of the transparent (invisible) surface. The inclination angle is estimated by seeking the value that leads to the minimal mutual information of the estimated scenes. The limitations and the consequences of noise and angle error are discussed, including a fundamental ambiguity in the determination of the plane of incidence. Experimental results demonstrate the success of angle estimation and consequent scene separation and labeling. PMID- 10680630 TI - High-diffraction-efficiency pseudorandom encoding. AB - Pseudorandom encoding (PRE) is a statistics-based procedure in which a pure-phase spatial light modulator (SLM) can yield, on the average, the prescribed diffraction pattern specified by the user. We seek to combine PRE with the optimization of an aperture-based target function. The target function is a fully complex input transmittance, unrealizable by a phase-only SLM, that generates a prescribed light intensity. The optimization is done to increase the diffraction efficiency of the overall process. We compare three optimization methods-Monte Carlo simulation, a genetic algorithm, and a gradient search-for maximizing the diffraction efficiency of a spot-array generator. Calculated solutions are then encoded by PRE, and the resulting diffraction patterns are computer simulated. Details on the complexity of each procedure are furnished, as well as comparisons on the quality, such as uniformity of the output spot array. PMID- 10680620 TI - Section 8--clinical relevance. American Institute of Ultrasound in Medicine. PMID- 10680631 TI - Propagation-invariant wave fields with finite energy. AB - Propagation invariance is extended in the paraxial regime, leading to a generalized self-imaging effect. These wave fields are characterized by a finite number of transverse self-images that appear, in general, at different orientations and scales. They possess finite energy and thus can be accurately generated. Necessary and sufficient conditions are derived, and they are appropriately represented in the Gauss-Laguerre modal plane. Relations with the following phenomena are investigated: classical self-imaging, rotating beams, eigen-Fourier functions, and the recently introduced generalized propagation invariant wave fields. In the paraxial regime they are all included within the generalized self-imaging effect that is presented. In this context we show an important relation between paraxial Bessel beams and Gauss-Laguerre beams. PMID- 10680632 TI - Dispersion control with use of long-period fiber gratings. AB - Chirped long-period fiber gratings are analyzed for management of dispersion in optical fiber communications systems. A ray model is used to derive simple analytic expressions that describe the transmission, chromatic delay, and dispersion properties of chirped long-period fiber gratings. A numerical model based on coupled-mode theory is used to verify the accuracy of the analytic expressions and explore design issues of the chirped long-period grating. With certain reasonable restrictions, chirped long-period gratings are found to be a viable and desirable alternative to existing dispersion compensation techniques. PMID- 10680633 TI - Elastic light scattering from single microparticles on a femtosecond time scale. AB - Experimentally measured and theoretically calculated elastic light-scattering spectra from single microparticles illuminated by 100-fs pulses are presented. Although in the theoretical calculation only a single incoming femtosecond laser pulse was used, the spectral behavior of scattered light shows all the features seen in the experimental spectrum from many femtosecond pulses, including morphology-dependent resonances (MDR's). The good agreement between experimental and theoretical elastic light-scattering data has stimulated a theoretical investigation of the time-dependent behavior of the elastically scattered light from a single microparticle on a femtosecond time scale. Since the spatial pulse length of the incoming laser pulse is smaller than the particle circumference, the temporal behavior of reflection, diffraction, refraction, and coupling into MDR's can be distinguished. Since the time-dependent scattering is strongly dependent on particle size, refractive index, and pulse chirp, it may be possible to encode several bits of information into a single laser pulse and therefore to increase optical data communication rates. PMID- 10680634 TI - Rigorous vector diffraction of electromagnetic waves by bidimensional photonic crystals. AB - We present a numerical study of bidimensional photonic crystals with an emphasis on the behavior of the gaps versus the polarization and the conicity of the incident plane wave. We use a rigorous modal theory of diffraction at oblique incidence by a set of arbitrarily shaped parallel fibers. This theory allows the study of the refractive properties of bidimensional photonic crystals. We develop a heuristic method of homogenization that allows us to predict the position of the gaps and their behavior with respect to the polarization and the conicity angle. With this homogenization scheme, we also present some important elements for obtaining full gaps. PMID- 10680635 TI - Characteristic properties of Mueller matrices. AB - A complete and minimum set of necessary and sufficient conditions for a real 4 x 4 matrix to be a physical Mueller matrix is obtained. An additional condition is presented to complete the set of known conditions, namely, the four conditions obtained from the nonnegativity of the eigenvalues of the Hermitian matrix H associated with a Mueller matrix M and the transmittance condition. Using the properties of H, a demonstration is also presented of Tr(M(T)M) = 4m(2)00 as being a necessary and sufficient condition for a physical Mueller matrix to be a pure Mueller matrix. PMID- 10680636 TI - Polarization of almost-plane waves. AB - The general polarization behavior of almost-plane waves, in which the electric field varies slowly over a circular pupil, is considered, on the basis of an axial Hertz potential treatment and expansion in Zernike polynomials. The resultant modes of a circular aperture are compared with the well-known waveguide (or optical fiber) modes and Gaussian beam modes. The wave can be decomposed into partial waves of electric and magnetic types. The modes for a square pupil are also considered. The particular application of the effect on polarization of focusing the waves is discussed. Another application discussed is the Fresnel reflection from a dielectric interface, it being shown that the Fresnel reflection alters the relative strength of the electric and magnetic components. PMID- 10680637 TI - Structure of the set of paraxial optical systems. AB - The set of paraxial optical systems is the manifold of the group of symplectic matrices. The structure of this group is nontrivial: It is not simply connected and is not of an exponential type. Our analysis clarifies the origin of the metaplectic phase and the inherent limitations for optical map fractionalization. We describe, for the first time to our knowledge, an image girator and a cross girator whose geometric and wave implementations are of interest. PMID- 10680638 TI - Aberration and the Strehl ratio. AB - An exact expression is derived for the Strehl ratio as a function of the minimum root-mean-square aberration value that may produce it. The result is applicable to any aperture and is shown to imply a similar expression for the Strehl ratio as a function of the minimum amplitude range of the aberration. PMID- 10680639 TI - The prevalence of C677T mutation in the methylenetetrahydrofolate reductase gene and its association with venous thrombophilia in Taiwanese Chinese. AB - C677T mutation of the methylenetetrahydrofolate reductase gene remains a controversial risk factor for venous thrombosis in Whites. The prevalence of methylenetetrahydrofolate reductase C677T genotype and its association with vascular thrombosis are not well established in Chinese population. We conducted a case-control study to investigate the prevalence of methylenetetrahydrofolate reductase C677T gene mutation and its association with venous thrombophilia in Taiwanese Chinese. The subjects consisted of 112 venous thrombophilic patients and 125 healthy controls, with similar age (p=0.08) and sex (p=0.58). The prevalent rates of C/T heterozygote were 32.8 and 44.6%; whereas those of T/T homozygote were 6.4 and 8.0% in the controls and patients, respectively. Neither C/T heterozygote (odds ratio, 1.7; 95% confidence interval, 1.0-3.0, p=0.05] nor T/T homozygote (odds ratio, 1.4; 95% confidence interval, 0.5-4.0, p=0.5) was significantly associated with venous thrombosis. Even when only subjects (52 patients and 107 controls) with normal inhibitor protein levels were analyzed, the association of T/T homozygote with venous thrombosis remained insignificant (p=0.06) with an odds ratio (95% confidence interval) of 3.4 (0.99-11.7). We concluded that, in Taiwanese Chinese, methylenetetrahydrofolate reductase C677T mutation is a common genetic mutation, but T/T homozygote is not a significant risk factor for venous thrombophilia. PMID- 10680640 TI - Factors affecting the first recurrence of noncardioembolic ischemic stroke. AB - Studies of the factors affecting the first recurrence of ischemic stroke have reported inconsistent findings. Types of initial stroke and the racial differences in study samples are among the explanations that may account for this inconsistency. The aims of this study were to estimate the cumulative recurrence rates of noncardioembolic ischemic stroke and identify the factors that influence the first recurrence of noncardioembolic ischemic stroke in the Taiwanese Chinese population. Four hundred and sixty-six patients with noncardioembolic ischemic stroke from thirteen hospitals in Taiwan were followed up in this study to ascertain first recurrence of noncardioembolic ischemic stroke between October 1992 and April 1995. The Kaplan-Meier method was used to estimate the cumulative recurrence rate. The Cox regression model was used to ascertain the significant factors affecting the first recurrence of noncardioembolic ischemic stroke. The overall cumulative recurrence rate was 10.5% (49/466) from the follow-up period of 30 months. After adjustment for age, sex, treatment modes, and variables pertinent to blood pressure, the site of brain lesion remained a significant factor. The relative risk of first recurrence for the basal ganglion vs. the region of middle cerebral artery was 3.06 (95% CI: 1.29-7.26). The brain lesion site was demonstrated to be an independent predictor of risk for the first recurrence of noncardioembolic ischemic stroke among the Taiwanese Chinese population. Whether this finding was also seen in other populations should be corroborated in future research. PMID- 10680641 TI - Markers of activated hemostasis and fibrinolysis in patients with pulmonary malignancies: comparison of plasma levels in central venous and pulmonary venous blood. AB - Malignancy frequently is accompanied by activated coagulation and fibrinolysis indicating a hypercoagulable state. The purpose of our study was to estimate the contribution of local tumor-induced mechanisms to the activation of hemostasis and fibrinolysis. In a prospective study, we compared the plasma levels of thrombin-antithrombin complexes, prothrombin fragment 1+2, and D-dimers in blood samples that simultaneously were drawn from the superior vena cava and the pulmonary vein of a tumor-bearing pulmonary lobe. Samples from the superior vena cava were drawn before operation and served as controls. After thoracotomy, a second group of samples was simultaneously taken from the pulmonary veins of the tumor-bearing lobe and the superior vena cava. Forty-five patients with pulmonary malignancies were included (25 adenocarcinomas and 20 squamous cell carcinomas). There were no significant differences of thrombin-antithrombin complexes, prothrombin fragment 1+2, and D-dimers levels in patients suffering from adenocarcinoma and from squamous cell carcinoma. Intraoperatively, prothrombin fragment 1+2 and D-dimers levels were markedly increased when compared with the preoperative values (p<0.0001). There was no increase of thrombin-antithrombin complexes levels due to the operative traumatization. Prothrombin fragment 1+2, thrombin-antithrombin complexes, and D-dimers plasma levels were significantly higher in the pulmonary venous blood than in the blood simultaneously drawn from the superior vena cava (p<0.0001). Our findings indicate that malignant lung tumors directly contribute to the activation of hemostasis and fibrinolysis in these clinical settings. PMID- 10680642 TI - NO reduces PMN adhesion to human vascular endothelial cells due to downregulation of ICAM-1 mRNA and surface expression. AB - Reperfusion damage is largely due to the adherence of polymorphonuclear leukocytes to the endothelium initiated by adhesion molecule upregulation. The reduced endothelial nitric oxide release during ischemia may be involved in the upregulation of intercellular adhesion molecule 1. In this study, we tested if nitric oxide donors suppress polymorphonuclear leukocyte adherence to activated endothelial cells by inhibition of the intercellular adhesion molecule 1 surface expression. Confluent human umbilical vein endothelial cells were stimulated with tumor necrosis factor alpha (300 U/mL) after preincubation with increasing concentrations of the nitric oxide donors CAS 1609 (0.005-5 mM/L) and 3-(4 morpholinyl)-sydnonimine (0.01-1 mM/L). Intercellular adhesion molecule 1 surface expression was measured in a cell surface enzyme-linked immunosorbent assay, intercellular adhesion molecule 1 mRNA by Northern analysis. Human saphenous vein endothelial cells were transfected with the inducible nitric oxide synthase gene and stimulated with tumor necrosis factor alpha (300 U/mL). Fluorescein green labeled polymorphonuclear leukocytes adhering to activated human umbilical vein endothelial cells/human saphenous vein endothelial cells were quantified by epifluorescent microscopy. The intercellular adhesion molecule 1 surface expression of activated human umbilical vein endothelial cells/human saphenous vein endothelial cells was significantly diminished to 40 to 60% of the maximum after treatment with CAS 1609, 3-(4-morpholinyl)-sydnonimine, or transfection with the inducible nitric oxide synthase gene. Intercellular adhesion molecule 1 mRNA was diminished by CAS 1609 and 3-(4-morpholinyl)-sydnonimine in the same manner. The functional relevance of our data was shown by reduction of polymorphonuclear leukocyte adherence to activated human umbilical vein endothelial cells/human saphenous vein endothelial cells following treatment with CAS 1609 and 3-(4-morpholinyl)-sydnonimine or transfection with inducible nitric oxide synthase. Tumor necrosis factor-induced polymorphonuclear leukocyte adherence was abolished by blocking antibody against intercellular adhesion molecule 1. Thus, exogenous or endogenous substitution of nitric oxide diminishes the expression of endothelial intercellular adhesion molecule 1 and its mRNA following tumor necrosis factor alpha stimulation. This results in a reduced polymorphonuclear leukocyte adherence to activated endothelium. PMID- 10680643 TI - Effect of DT-TX 30, a combined thromboxane synthase inhibitor and thromboxane receptor antagonist, on retinal vascularity in experimental diabetes mellitus. AB - Combined thromboxane synthase inhibitors and thromboxane receptors antagonists have been shown to have a beneficial effect on different models of thrombosis in vivo. We studied the action of one of these compounds (DT-TX 30) compared with dazoxiben (a thromboxane synthase inhibitor) on retinal vascularity in streptozotocin-diabetic rats. Ten nondiabetic animals were treated with isotonic saline, 30 (10 animals per group) were given 0.4, 4, and 8 mg kg(-1) per day of DT-TX 30 (p.o.) and 30 (10 animals per group) were given 10, 50, and 100 mg kg( 1) per day of dazoxiben (p.o.) over a 90-day study period. DT-TX 30 caused a dose dependent decrease of platelet aggregation and thromboxane B2 synthesis. There was an increase of 9, 65, and 166% in the synthesis of prostacyclin after treatment with 0.4, 4, and 8 mg kg(-1) per day, respectively. Retinal vascularity increased in 51, 72, and 182% of animals in response to the three doses used. Synthesis of prostacyclin and the degree of retinal vascularity showed a linear correlation (r2=0.6528,p<0.00001). Dazoxiben, at doses that inhibited thromboxane synthesis as much as DT-TX 30, increased prostacyclin production and retinal vascularity with less potency than DT-TX 30. In conclusion, the antagonism of thromboxane receptors may be an important additional effect to the inhibition of thromboxane synthase in the prevention of ischemic retinal lesions in experimental diabetes. PMID- 10680644 TI - A recombinant antibody-targeted plasminogen activator with high affinity for activated platelets increases thrombolytic potency in vitro and in vivo. AB - To increase thrombolytic specificity of urokinase (uPA), we engineered a recombinant chimeric plasminogen activator SZ51Hu-scuPA, which consists of a humanized monoclonal antibody (SZ-51Hu) specifically against P-selectin on activated human platelet and a single-chain urokinase (scuPA). The cDNA, encoding scuPA amino acids 1-411, was inserted in 5' end to 3' end orientation immediately after the CH3 of SZ-51Hu heavy-chain sequence in the expression vector alphaLys30. The resulting construct alphaLys30-SZ51VH/Hu-scuPA was used to transfect into SP2/0 murine myeloma cell line, which was pretransfected with SZ51Hu light chain. The fusion protein SZ51Hu-scuPA was expressed at 5 mg/L in the supernatant of cell culture. The fusion protein purified by affinity chromatography had a molecular weight of 160 kDa with fibrinolytic activity of 39,000 IU/mg and its affinity to activated human platelet was 67% of the parent murine mAb SZ-51. The thrombolytic property of the fusion protein was first characterized in an in vitro system, which consists of a 125I-fibrin-labeled human plasma clot containing different concentrations of human platelets suspended in citrated human plasma. Fifty percent lysis was reached with SZ51Hu scuPA in 1 hour at a concentration of 20 IU/mL or in 2 hours at a concentration of 10 IU/ mL, which was much faster than uPA at the same concentration. The maximal lysis of the clots by SZ51Hu-scuPA was 4.1 to 8.4 times more potent than that by uPA. The fusion protein was further characterized in the hamster pulmonary embolism model with clots prepared from fresh platelet-rich human plasma containing 125I-labeled fibrinogen. The thrombolytic activity of SZ51 scuPA was 3.9 times more potent than that of uPA at 2,000 IU/kg in this model. Almost no significant fibrinogen breakdown was observed either in vitro and in vivo. PMID- 10680645 TI - Platelet aggregation during abdominal surgery in an experimental pig model: the effects of presurgical antibiotic protocols and volume replacement with hydroxyethyl starch. AB - The effect of presurgical antibiotic protocols in combination with hemodilution on platelet aggregation was studied. Thirty pigs were randomly assigned to three groups. Group 1 received amoxicillin/clavulanic acid, group 2 metronidazole+cefuroxime, and group 3, as a control, sodium chloride. They underwent laparotomy, massive blood loss, and volume replacement with hydroxyethyl starch 200, followed by an anaphylactoid reaction. Platelet aggregation was measured by the turbidometric method. Neither antibiotic protocols had any effect on platelet aggregation as compared with the control group. In all three groups, aggregation to ADP and collagen was significantly reduced after volume replacement with hydroxyethyl starch. In contrast, the sensitivity to the aggregating effects of collagen was increased as assessed by a higher frequency of responses to low concentrations of collagen and a shortened latency of the aggregation response after collagen addition. Further in vitro studies revealed that dilution of plasma with hydroxyethyl starch specifically induced the changes seen after in vivo volume replacement. The results suggest that the plasma substitute hydroxyethyl starch 200 increases the sensitivity to low doses of collagen, an effect never described before and considered of clinical relevance. PMID- 10680646 TI - Detection of mononuclear cells as the source of the increased tissue factor mRNA in the liver from lipopolysaccharide-treated rats. AB - Tissue factor (TF) triggers the coagulation cascade reaction in vivo. Overexpression of TF mRNA is one leading cause of disseminated intravascular coagulation and thrombosis-related organ failure. In response to lipopolysaccharide (LPS) stimulation, various cell types can produce TF mRNA in vitro. However, there is currently no agreement on what types of cells in the liver overexpress TF mRNA after LPS treatment. For the first report, we found the increased TF mRNA with reverse transcription-polymerase chain reaction (RT-PCR), and confirmed a fourfold increase (p<0.001 vs. control, t-test) of the TF mRNA level with RT-competitive PCR in the liver of LPS-treated (2.0 mg/kg i.v. injection) rats. There was no significant difference in the glyceraldehyde-3 phosphate dehydrogenase mRNA level between LPS-treated rats and control rats. To clarify the localization and cellular source of LPS-induced TF mRNA, we performed in situ hybridization analysis with [35S]-labeled oligonucleotides probes, which we originally designed. We detected intense signals of TF mRNA in mononuclear cells but not in endothelial cells around the hepatic vein of LPS-treated rats. In this study, we showed that the TF mRNA level induced by LPS treatment, which may indicate mononuclear cells associated, significantly increased in the liver of rats. These results will provide circumstantial support for the therapeutic strategy that mononuclear cell should be one of the target cells to be treated in the early phase of disseminated intravascular coagulation in the liver, and that the need to suppress its overexpression of TF mRNA is essential for preventing hypercoagulable condition. PMID- 10680647 TI - Effect of methylenetetrahydrofolate reductase 677 C-T, 1298 A-C, and 1317 T-C on factor V 1691 mutation in Turkish deep vein thrombosis patients. AB - Possible effect of three common mutations in (MTHFR 677 C-T; 1317 T-C; 1298 C-A) and FV 1691 G-A mutation was studied in Turkish patients with thrombosis and compared with normal controls. The case-control study included 68 patients with the diagnosis of deep vein thrombosis and 66 controls, consecutively selected among subjects without personal and familial history of atherothrombosis. Patients with deep vein thrombosis were selected if Doppler ultrasonography was positive. Only, the comparison of factor V 1691 G-A mutation revealed statistically significant difference in control (6.06%) and deep vein thrombosis (23.5%) group. Risk assessment of double prothrombotic gene alterations revealed only FV 1691 G-A mutation as an independent risk factor for thrombosis (odds ratio 4.7 [1.5-15.0]), but our data suggested that MTHFR 677 has effect on its own (odds ratio 1.97 [0.6-2.7]) but may have synergy with FV 1691 G-A (odds ratio 8.12 [2.0-25.3]). However, MTHFR 1298 A-C and 1317 T-C does not have any effect; furthermore, being heterozygote at two different loci or homozygosity at least in a locus for 677 and 1298 revealed a significant increase (odds ratio 9 and 24 [1.3-59.3 and 2.3-240.3]) between these two groups. PMID- 10680648 TI - Free radicals are involved in cancer procoagulant-induced platelet activation. PMID- 10680649 TI - Vaccination of neonatal colostrum-deprived calves against Pasteurella haemolytica A1. AB - Colostrum-deprived Holstein calves were vaccinated at 2 and 4 wk of age with a Pasteurella haemolytica A1 culture supernatant vaccine to determine whether active immune responses and protection could be induced in this age group in the absence of maternal antibodies. All calves responded to vaccination with high titers of IgM antibodies to capsular polysaccharide within 1 wk of primary vaccination. Mean titers of IgG1 and IgG2 antibodies to this antigen increased significantly by 2 wk after secondary vaccination, but peak antibody titers were low. All of the vaccinated calves seroconverted with production of leukotoxin neutralizing antibodies, but peak antibody titers were low. Vaccinated calves experienced considerable lung damage after experimental challenge, but survival rate, clinical scores, and percent lung involvement were significantly better than those of control (placebo-injected) calves. PMID- 10680650 TI - Prevention of edema disease in pigs by passive immunization. AB - The effect of treatment with verotoxin 2e (VT2e) specific antiserum was evaluated in 3 Danish pig herds with edema disease (ED). The antiserum was prepared by immunizing horses with a VT2e toxoid. The study was performed as a randomized blind field trial with parallel treatment and control groups. There were approximately 50 piglets in each group in each of the 3 herds and 741 piglets were included in the study (244 from herd A, 249 from herd B, and 247 from herd C). Treatment groups received 2, 4, or 6 mL anti-VT2e serum intramuscularly the day before weaning. Control groups were treated with 6 mL normal horse serum or 6 mL RPMI 1640 medium as placebo. All pigs that died in the trial period (1 d before weaning to 44 d after weaning) were examined pathologically and microbiologically. Mortality due to ED, mortality due to other causes, and adverse effects due to treatment were recorded. As there was no mortality due to ED, herd B was excluded from statistical calculations on mortality. The content of horse antibodies specific to VT2e in serum from pigs was analyzed in an indirect ELISA. A higher dose of anti-VT2e serum was reflected in higher optical density values in the indirect ELISA. Transient adverse reactions, seen as vomiting, ataxia, and cyanosis, occurred shortly after the injection of horse serum in 1.5% of the pigs, and one pig died. There were no statistically significant differences in mortality due to other causes among the 3 treatment groups in herds A and C. Only pigs from which F18+, VT2e+, ST-, LT- hemolytic E. coli (0139 or O-rough) was isolated were diagnosed as dead due to ED. Deaths due to ED in the control groups were 8.1% and 12.0% in herds A and C, respectively, compared with 0% and 0.7% in the corresponding serum groups. The difference between treatment and control groups was statistically significant (P<0.0001). It was not possible to establish an effect of dose (2, 4, or 6 mL) of anti-VT2e serum, because only one pig died of ED in the treatment groups. It was concluded that passive immunization by intramuscular injection of a VT2e-specific antiserum can be used for protecting piglets against ED. PMID- 10680651 TI - Subtypes of intimin among non-toxigenic Escherichia coli from diarrheic calves in Brazil. AB - One hundred and five strains of Escherichia coli that were isolated from calves with diarrhea in the state of Sao Paulo, Brazil, and were negative for enterotoxins and cytotoxins, were examined for the eae gene. Four (3.8%) strains were positive by polymerase chain reaction (PCR) and were shown to produce intimin by using Western blot with specific antiserum against the conserved N terminal region of intimin. Subtyping of the intimins was done by PCR with specific primers and by Western blot with specific antisera against the C terminal variable region of the protein. Three of these isolates (O?:H11, O26:H-, O123:H1) produced the beta subtype of intimin, and the 4th (0103:H2) produced intimin that was not typable. The 0103:H2 and the O26:H-isolates adhered to HEp-2 cells with diffuse adherence and localized-like adherence patterns, respectively. The other strains did not adhere to HEp-2 cells. To our knowledge, this is the first report of the occurrence of a subtype of intimin described for human enteropathogenic E. coli among bovine diarrheogenic E. coli. It is also the first report from Brazil demonstrating the presence of bovine E. coli harboring the eae gene. PMID- 10680652 TI - Experimental aerosol transmission of Actinobacillus pleuropneumoniae to pigs. AB - In order to demonstrate the possible role of aerosol in the transmission of Actinobacillus pleuropneumoniae, an experiment including 18 specific pathogen free (SPF), 10-week-old piglets, randomly distributed into 2 adjacent units, was carried out. In these facilities, air was forced through absolute filters to prevent any contact with infectious agents. During the first 6 d post inoculation, the 2 units were connected by a rectangular opening and the air circulation was forced by the ventilation system from unit A (inoculated pigs) to unit B (non-inoculated pigs). The A. pleuropneumoniae strain (biovar 1 serovar 9) was isolated in France from an outbreak of porcine pleuropneumonia. Two different infecting doses, 10(7) cfu/animal and 10(8) cfu/animal, were inoculated by intranasal route in 6 pigs of unit A. The infection spread quickly from the inoculated pigs to the non-inoculated pigs. Clinical signs were acute during the 4 d post inoculation: hyperthermia, respiratory distress and, sometimes, death (6 pigs of the unit A and 2 pigs of the unit B). All pigs seroconverted against A. pleuropneumoniae serovar 9 within 2 weeks. Lung lesions were severe: fibrinous pleurisy and lung hemorrhages in the acute stage, pleural adherences and focal pulmonary necrosis in the chronic stage. Actinobacillus pleuropneumoniae was isolated from the tonsils and/or lungs in 16 animals. It could be also isolated from the air of the experimental unit. This study showed that A. pleuropneumoniae was readily transmitted through aerosol over a distance of at least 2.5 m. PMID- 10680653 TI - Assessment of various treatments to reduce carriage of Salmonella in swine. AB - In this study, different strategies to reduce carriage of Salmonella spp. in pigs were evaluated. Probiotics, prebiotics, vaccination, and acidification of drinking water were assessed as means of reducing Salmonella. Acidification of water, use of egg yolk-specific immunoglobulins, and vaccination with an endotoxin vaccine did not reduce Salmonella excretion in experimentally infected pigs. A reduction of Salmonella in the colonization of mesenteric lymph nodes was observed with the use of bambermycins and a live attenuated vaccine. A reduction in the shedding of S. Typhimurium was also observed after supplementation with fructooligosaccharides in drinking water. The use of probiotics and prebiotics appeared to change the pig fecal bacterial flora as indicated by Gram staining of smears from rectal swabs. PMID- 10680654 TI - Development of an indirect enzyme-linked immunosorbent assay for the detection of leptospiral antibodies in dogs. AB - Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources, expertise, or facilities to perform the microscopic agglutination test (MAT). Thus, there is a need for a rapid and simple serological test that could be used in any diagnostic laboratory. In this study, a genus-specific, heat-stable antigenic preparation from Leptospira interrogans serovar pomona was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of leptospiral antibodies in dog sera. This antigenic preparation reacted with rabbit antisera against L. interrogans serovars bratislava, autumnalis, icterohaemorrhagiae and pomona and with rabbit antiserum against L. kirschneri serovar grippotyphosa. The ELISA showed a relative specificity of 95.6% with 158 dog sera which were negative at a dilution of 1:100 in the MAT for serovars pomona, bratislava, icterohaemorrhagiae, autumnalis, hardjo, and grippotyphosa. The relative sensitivity of this assay with 21 dog sera that revealed serovars MAT titres of > or =100 to different serovars was 100%. This assay is easily standardized, technically more advantageous than MAT, and uses an antigenic preparation that can be routinely prepared in large amounts. It was concluded that this ELISA is sufficiently sensitive test to be used as an initial screening test for the detection of leptospiral antibodies in canine sera, with subsequent confirmation of positive test results with the MAT. PMID- 10680655 TI - Detection of antibodies to equine arteritis virus by a monoclonal antibody-based blocking ELISA. AB - A potent ELISA antigen was prepared from equine arteritis virus (EAV) by differential centrifugation of EAV-infected cell culture fluid, followed by solubilization of the preparation by Triton X-100 treatment. Using this antigen and a mouse monoclonal antibody against the G(L) protein of EAV, a reliable blocking ELISA (bELISA) was developed for the detection of EAV antibodies in equine sera. The bELISA was evaluated using a total of 837 test serum samples. The relative sensitivity (n = 320) of the bELISA compared to the serum neutralization (SN) test was 99.4%. The bELISA appears to be a highly specific test, the specificity of which did not appear to be adversely affected by previous exposure of horses to non-EAV-containing biologicals. Of 119 serum samples, 21 from horses without any history of exposure to EAV and 98 from racetrack Thoroughbreds, 118 were negative in the SN test and bELISA. One sample was SN-negative but suspicious with the bELISA. Based on testing 465 SN-negative field samples and 52 SN-negative samples from experimental horses, and excluding any sera giving a suspicious reaction, the relative specificity of the bELISA was 97.7%. Samples should be examined undiluted and diluted 1/10 in the bELISA because the testing of sera of high neutralizing antibody titer may be affected by a prozone-like phenomenon. The bELISA is a more rapid and cost-efficient test than the SN test for the detection of EAV antibodies in equine sera. PMID- 10680656 TI - PCR detection and characterization of type-2 porcine circovirus. AB - A polymerase chain reaction (PCR) assay was developed for detecting porcine circovirus (PCV). The assay readily detected type-2 PCV (PCV-2) and type-1 PCV (PCV-1). The PCR primers were designed based on DNA sequences conserved in all reported PCV genomes. Type 1 PCV and type 2 PCV both produced 438 bp amplification products, which were easily identified and differentiated from one another by restriction fragment length polymorphism (RFLP) analysis. Porcine circovirus was detected in 55% (931/1693) of randomly tested pigs with various clinical signs and lesions, most of which were difficult to differentiate from those associated with porcine reproductive and respiratory syndrome (PRRS). The PCR products from all positive clinical samples were identified by RFLP to be only PCV-2; DNA tested by PCR was extracted directly from one or more of lung, mesenteric or mediastinal lymph nodes, and tonsil. Type 2 PCV was also detected in 6% (2/34) of DNA extracted directly from semen of randomly chosen healthy boars. Positive PCR reactions from 554 diseased pigs were characterized by RFLP and categorized into 5 different profiles (A-E), of which 82.8% were PCV-2A (456/554), 3.0% were PCV-2B (17/554), 9.9% were PCV-2C (55/554), 1.1% were PCV-2D (6/554), and 3.2% were PCV-2E (18/554). The complete genomic nucleotide sequences of PCV-2A, B, C, D, and E were determined and found to have at least 95% homology compared with one another and with all other PCV-2 found in the GenBank database. All PCV-2 had less than 76% homology with PCV-1. This PCR assay will hopefully be useful to veterinary diagnostic laboratories for routine testing and surveillance of infection with PCV-2. The RFLP profiling system might be useful for preliminary characterization and identification of PCV isolates and might also benefit studies on the molecular epidemiology of PCV. PMID- 10680657 TI - Effect of aflatoxin on performance, hematology, and clinical immunology in lambs. AB - Twenty-four female lambs were intoxicated with a diet contaminated with 2 ppm aflatoxin for a period of 37 d. Twelve lambs were maintained as the control group. After this period, the lambs were left for 35 d without aflatoxin in their feed. Performance, hematology and clinical immunology were examined in the intoxicated lambs. A non-significant decrease in body weight was observed in the intoxicated lambs during the intoxication period, whereas a significant decrease (P<0.001) in average daily gain was noted on the last day of intoxication and during the clearance period. No significant differences were observed in erythrocyte count, white blood cell count or differential leukocyte count between the groups. Bacteriostatic activity of the serum was lower in the intoxicated lambs, however, there was no effect on serum opsonic activity. Phagocytosis by the neutrophils was higher during the intoxication period and the levels of IgG were elevated in the intoxicated lambs. In vivo cellular immunity was assessed by intradermal injection of phytohemagglutinin; the response was lower during intoxication period. These results indicate that a lowering in the average daily gain was the most sensitive indicator of aflatoxicosis in lambs, and that the immune response was altered, which could render the animals more susceptible to infectious diseases. PMID- 10680658 TI - Correlation of periovulatory serum and fecal progestins in the domestic dog. AB - This study was initiated to determine the relationship between canine ovarian steroids detected in serum and feces during the periovulatory interval in domestic dogs, and to examine the feasibility of a non-invasive method to estimate the time of ovulation in canid species. When bitches (n = 14) were observed to enter proestrus (based on vulvar enlargement or serosanguineous vaginal discharge), paired daily serum and fecal samples were collected for a 15- to 20-day period and stored at -20 degrees C. After extraction, progestin concentrations in both substrates were measured using an established enzyme immunoassay procedure. All samples were aligned to Day 0, the first day in which fecal progestins reached a sustained rise above 100 ng/g feces. Mean fecal progestin concentrations increased in parallel with mean serum progesterone values (r = 0.78), rising from 44.6+/-2.6 ng/g feces to 409.6+/-90.9 ng/g feces, and 5.4+/-0.9 nmol/L to 81.2+/-18.5 nmol/L, on Day -5 and Day 5, respectively. Individual fecal progestin concentrations varied markedly, but plotted against serum progesterone concentrations demonstrated correlation coefficients ranging from 0.41 to 0.97 (P<0.05). These results demonstrate that sequential changes in domestic dog serum ovarian steroid concentrations are paralleled in the feces, and that it is feasible to non-invasively monitor individual progestin changes in the periovulatory interval using fecal hormone analysis. PMID- 10680659 TI - Comparative cardiopulmonary effects of carfentanil-xylazine and medetomidine ketamine used for immobilization of mule deer and mule deer/white-tailed deer hybrids. AB - Three mule deer and 4 mule deer/white-tailed deer hybrids were immobilized in a crossover study with carfentanil (10 microg/kg) + xylazine (0.3 mg/kg) (CX), and medetomidine (100 microg/kg) + ketamine (2.5 mg/kg) (MK). The deer were maintained in left lateral recumbency for 1 h with each combination. Deer were immobilized with MK in 230+/-68 s (mean +/- SD) and with CX in 282+/-83 seconds. Systolic, mean and diastolic arterial pressure were significantly higher with MK. Heart rate, PaO2, PaCO2, pH, and base excess were not significantly different between treatments. Base excess and pH increased significantly over time with both treatments. Both treatments produced hypoventilation (PaCO2 > 50 mm Hg) and hypoxemia (PaO2 < 60 mm Hg). PaO2 increased significantly over time with CX. Body temperature was significantly (P<0.05) higher with CX compared to MK. Ventricular premature contractions, atrial premature contractions, and a junctional escape rhythm were noted during CX immobilization. No arrhythmias were noted during MK immobilization. Quality of immobilization was superior with MK, with no observed movement present for the 60 min of immobilization. Movement of the head and limbs occurred in 4 animals immobilized with CX. The major complication observed with both of these treatments was hypoxemia, and supplemental inspired oxygen is recommended during immobilization. Hyperthermia can further complicate immobilization with CX, reinforcing the need for supplemental oxygen. PMID- 10680660 TI - Effects of vitamins A and E on superoxide production and intracellular signaling of neutrophils in Holstein calves. AB - The effects of vitamin A and vitamin E treatment on superoxide (O2-) production of neutrophils and their intracellular signaling, including intracellular Ca2+, protein kinase C (PKC), and tyrosine kinase (TK), were examined in Holstein calves. After treatment with vitamin A, heat-aggregated IgG (H-agg.IgG)-induced O2- production in neutrophils ranged from 3.7+/-0.4 to 4.3+/-0.9 nmol (mean +/- SD), and it was significantly (P<0.05) higher than that in the control calves. Opsonized zymosan (OPZ)-induced O2- production was similar with that of control calves. Intracellular signaling of neutrophils in vitamin A-treated calves was enhanced compared with that of control calves when stimulated with H-agg. IgG, but not with OPZ. After treatment with vitamin E, H-agg.IgG- and OPZ-induced O2- production of neutrophils ranged from 4.2+/-0.8 nmol to 5.4+/-0.5 nmol and from 7.8+/-0.6 nmol to 8.1+/-0.4 nmol (mean +/- SD), respectively, and they were significantly (P<0.05) higher than those in control calves. Intracellular signaling was also enhanced compared with that of control calves. These results suggested that the effects of vitamin A and E treatment on O2- production were correlated to changes in intracellular signaling of neutrophils in Holstein calves. PMID- 10680661 TI - Does portal hypertension contribute to the pathogenesis of gastric ulcer associated with liver cirrhosis? AB - The prevalence of gastric ulcers in patients with liver cirrhosis is increased compared with that in the general population, and portal hypertension may contribute to the increased risk of gastric ulcer in cirrhosis patients. Aggressive factors involved in the pathogenesis of gastric ulcer are diminished in association with portal hypertension. In contrast, most of the important gastric mucosal defense mechanisms are shown to be impaired in portal hypertension; many of these mechanisms are also found to be altered in patients with liver cirrhosis. Portal hypotensive treatment with propranolol reduces ethanol-induced gastric mucosal damage in portal hypertensive rats and improves endoscopic signs of portal hypertensive gastropathy in cirrhosis patients. Together, these findings suggest portal hypertension-induced impairment of the gastric mucosal defenses to be an important factor in the pathogenesis of gastric ulcer in patients with liver cirrhosis. Prospective studies of portal pressure reducing procedures, such as pharmacotherapy with propranolol, and their effect on the incidence of gastric ulcer in cirrhosis patients are needed to confirm this suggestion. PMID- 10680662 TI - Feasibility of optical coherence tomography for high-resolution imaging of human gastrointestinal tract malignancies. AB - Optical coherence tomography (OCT) is a new imaging technology which can perform high-resolution, cross-sectional imaging of the internal microstructure of biological tissues. OCT is analogous to ultrasound, except that it measures the intensity of back-reflected infrared light rather than sound waves. OCT performs two- and three-dimensional imaging of tissue microstructure in situ and in real time. It can achieve image resolutions approaching the cellular level over approximately the same imaging depths as a conventional biopsy. In this article we examine the feasibility of OCT for high-resolution imaging of gastrointestinal malignancies with ex-vivo imaging of normal and pathologic microstructures. Tissue, both normal and neoplastic, was obtained from patients undergoing surgical resection after an initial diagnosis of a gastrointestinal malignancy. The tissue samples were imaged prior to fixation using a laboratory OCT system. The OCT system consists of a fiber optic-based Michelson interferometer, a commercially available amplified superluminscent light source, and a computer for data acquisition. The images were subsequently compared with histological cross sections corresponding to the imaged areas. The stratified squamous epithelium of the normal esophagus was clearly visible in the OCT images and contrasted to the disorganized and non-uniform nature of the mucosal layers of Barrett's esophagus and squamous carcinoma. The columnar epithelial morphology as well as other mucosal structures in normal colon were distinctly visible using OCT. In contrast, disorganization of the normal mucosal layers and ulcerative lesions were identified in tissues from ulcerative colitis and adenocarcinoma of the colon. The ability of OCT to image tissue microstructure at high resolutions makes it a potentially powerful technology for minimally invasive assessment of the gastrointestinal tract and the evaluation of early neoplastic changes. PMID- 10680663 TI - Altered vascular response to acetylcholine in conditions of endothelial damage in the isolated perfused rat stomach. AB - To examine the mechanism of stress ulcers and the relation between endothelium derived relaxing factor (EDRF)/NO and gastric mucosal blood flow (GMBF), we used an isolated perfused rat stomach model and studied the effects of an autonomic nerve activator, nitric oxide synthase (NOS) synthesis, and an EDRF/NO inhibitor on gastric blood circulation. Rats were divided into four groups according to pretreatment: (1) control; (2) those given gossypol, a drug provoking endothelial cell damage; (3) those given L-N monomethylarginine (L-NMMA), a specific inhibitor of EDRF/NO; and (4) those subjected to water-immersion stress. Using this model we collected the perfusion fluid from the portal vein at various time points. After administration of acetylcholine, the perfusion flow increased in the control group, but perfusion flow showed no change in the L-NMMA group. On the other hand, the perfusion flow decreased in the gossypol and water-immersion stress groups. The perfusion fluid from the control group contained cGMP, but this substance was absent in the perfusion fluid of the other experimental groups. We considered that increased cGMP in the fluid came from endothelial cells. We presume that the presence of EDRF/NO is essential for the control of GMBF and that from the viewpoint of gastric ulcers, the lack of EDRF/NO may be an important factor in the decrease of GMBF in the early stages of water-immersion stress. PMID- 10680664 TI - Clinical importance of n-3 fatty acid-rich diet and nutritional education for the maintenance of remission in Crohn's disease. AB - Elemental diet (ED) therapy has been established as primary therapy for Crohn's disease, and home enteral nutrition (HEN) has been reported to control relapse at a dose of more than 30kcal/kg of ideal body weight. However, a decrease in ED compliance with long-term use is becoming problem. We developed an n-3 fatty acid rich diet and carried out nutritional education specifically for Crohn's disease patients using HEN to facilitate compliance and to improve their nutritional status. After the introduction of this n-3 rich diet, disease activity was not altered, and nutritional status, especially serum n-3 fatty acid levels, improved. The remission periods in patients with poor compliance seemed to be prolonged by the nutritional education. Thus, a n-3 rich diet in combination with nutritional education specific for Crohn's disease patients is very important for the in maintenance of high compliance and for maintaining nutritional balance. PMID- 10680665 TI - Changes of liver fibrosis in chronic hepatitis C patients with no response to interferon-alpha therapy: including quantitative assessment by a morphometric method. AB - The aim of this study was to evaluate the antifibrotic effect of interferon (IFN) alpha in chronic hepatitis C (CH-C) patients with no response to IFN-alpha therapy. We studied 76 patients (46 men, 30 women; mean age, 55.6 years) who received IFN-alpha intramuscularly, at a total close of 480 to 880MU for 6 months (group A). As a control group, we studied 50 patients (32 men and 18 women; mean age, 58.5 years) with CH-C who received medication other than IFN (ie, Strong-Neo Minophagen C, ursodeoxycholic acid, and a herbal medicine, Sho-saiko-to [TJ-9]) and who had persistent alanine aminotransferase (ALT) elevation (group B). All patients were subdivided into three subgroups according to different patterns of ALT changes during the observation period, ie, (a) persistent ALT level < 60IU/ 1 (below about twice the upper limit of the normal range), (b) persistent ALT level > or = 60IU/1, (c) ALT levels other than (a) and (b). Liver biopsy was performed within 6 months prior to IFN therapy and more than 6 months after IFN therapy, while two liver biopsies were performed during therapy in group B. Liver fibrosis was compared between two specimens by staging. When the fibrosis stage was the same in the two specimens, we determined whether the fibrosis had improved or worsened by comparing the fibrotic ratio, ie, the ratio of the area of fibrosis to the area of the entire liver tissue specimen, calculated using computed graphic software. Serum aminoterminal peptide of type III procollagen (PIIIP) levels were measured on the day of the liver biopsy and their mean yearly changes were compared between the two groups. Improvement of liver fibrosis was found in 12% to 30% of patients in each ALT subgroup and in 24% of all patients in group A and there were no significant differences in liver fibrosis in comparison with findings in of group B when assessed by staging alone. However, these percentages rose to 59% to 75% and 66%, respectively, when liver fibrosis was assessed by the fibrotic ratio together with staging, resulting in a significant difference in fibrosis between groups A and B in total (P < 0.01). The mean yearly changes in serum PIIIP levels in each subgroup and in all patients in group A were below zero, indicating a tendency to improvement of fibrosis after IFN therapy, while these changes in group B were all above zero, except for subgroup (c). Improvement of fibrosis after IFN therapy was found in 15 of 24 patients (64%) whose ALT changes had the same pattern before and after IFN therapy, although no significant difference was noted between improved and worsened patients. These results suggest that IFN-alpha may have an antifibrotic effect even in CH-C patients with no overt response to IFN-alpha therapy, compared with the effect of medications other than IFN. PMID- 10680666 TI - Augmenter of liver regeneration (ALR) may promote liver regeneration by reducing natural killer (NK) cell activity in human liver diseases. AB - Cytotoxicity of liver natural killer cells against regenerating hepatocytes has been reported as a possible mechanism of regeneration failure in fulminant hepatitis. An augmenter of liver regeneration (ALR) inhibits liver natural killer cell activity in rats. In this study, we measured hepatic expression of ALR mRNA, blood levels of ALR, and peripheral blood natural killer cell activity in patients with various types of acute liver disease to investigate the relationship between failure of liver regeneration and hepatic natural killer cells. Hepatic ALR mRNA expression was higher in liver disease patients than in non-liver disease controls, and a correlation was found between serum ALR values and hepatic levels of ALR mRNA. In acute liver injury, the serum ALR level also showed a negative correlation with NK activity. ALR was produced by and released from the liver at the time of hepatic injury. Our findings suggest that ALR may protect against failure of regeneration by inhibition of hepatic natural killer cell activity in acute liver injury. PMID- 10680667 TI - Optimal route of administration of mixed endothelin receptor antagonist (TAK-044) in liver transplantation. AB - It is well known that endothelin-1(ET-1) is a factor involved in the pathogenesis of ischemia-reperfusion injury. This study was undertaken to investigate the optimal route (intravenous vs intraportal) for administering mixed endothelin receptor antagonist (TAK-044) in a liver transplantation. First, in a rat isolated liver cold-perfusion model, the pharmacodynamics of TAK-044 and endothelin-1 (ET) in the liver tissue and the systemic circulation after cold perfusion were compared in the different administration routes. Next, in a rat orthotopic transplantation model, we compared the hepatoprotective effect of TAK 044 among different administration routes. In each model, there were three groups: IV group, intravenous injection of TAK-044 (10mg/kg) immediately before cold perfusion or anhepatic phase; IP group, intraportal administration with cold perfusion solution or with reflush solution for the graft; control group, no treatment. In the cold perfusion model, liver tissue ET level increased to a similar extent after reperfusion in the three groups, and the plasma and liver tissue TAK-044 concentrations after reperfusion were highest in the IV group. However, the increase in plasma ET was also greatest, and therefore, the ratio of liver tissue to plasma TAK-044 was lower in the IV group compared with the IP group. In the transplantation model, elevation of plasma ET was significantly higher in the IV group. Leakage of serum alanine aminotransferase (ALT), sinusoidal narrowing, and cell swelling after grafting were significantly suppressed in the IP group. We conclude that intraportal administration before reperfusion offers more efficient accumulation of TAK-044 in the liver tissue, without harmful systemic elevation of ET, and achieves a hepatoprotective effect on the graft compared with intravenous administration. PMID- 10680668 TI - Early diagnosis of interferon-induced myocardial disorder in patients with chronic hepatitis C: evaluation by myocardial imaging with 123I-BMIPP. AB - Interferon (IFN) therapy for chronic hepatitis C is sometimes associated with cardiac complications. In the present study, we performed myocardial imaging with 123I-labeled beta-methyl-p-iodophenylpentadecanoic acid (123I-BMIPP) in order to evaluate myocardial disorders caused by IFN. We studied 40 healthy subjects (H group) and 25 patients with chronic hepatitis C who had been treated with IFN (IFN group). A Holter electrocardiogram (ECG) was performed and the autonomic nervous function was assessed by analyzing the spectral variability and 1/f fluctuation of heart rate. Myocardial planner imaging with 123I-BMIPP was performed to obtain the time activity curve for 20min immediately after administration of 123I-BMIPP (dynamic study). Early and delayed myocardial single photon emission computed tomography (SPECT) images were expressed as Bull's eyes and the myocardium was divided into four segments to calculate the washout rate for each segment on early and late SPECT images (early and late SPECT study). No significant differences in autonomic nervous function were observed between the two groups in heart rate variability. In a dynamic study, the reduction rate from the time activity curve was significantly higher in the IFN group compared with the H group (reduction rate, IFN group, 5.3 +/- 3.7% vs H group, 1.2 +/- 3.3%; P < 0.05). In the early and delayed myocardial SPECT study, the washout rate for the IFN group was significantly increased in all myocardial areas compared to that in the H group. However, the metabolic disorder of fatty acids caused by IFN was reversed on the second 123I-BMIPP myocardial scintigraphy examination several months after IFN therapy. These results indicate that metabolic disorders of fatty acids caused by IFN therapy can be detected before abnormalities are observed by Holter-ECG or echocardiography. PMID- 10680669 TI - Nationwide epidemiological survey of chronic pancreatitis in Japan. AB - The aim of this study was to estimate the number of patients treated for chronic pancreatitis in 1994 in Japan and to explore the clinico-epidemiological features of chronic pancreatitis. Two surveys were conducted. Stratified random sampling was used to select departments in which patients with chronic pancreatitis were treated, and two different questionnaires were administered to obtain relevant information. From the first survey, the total number of patients treated for chronic pancreatitis in Japan in the year 1994 was estimated as 32,000 (95% confidence interval, 25,000-39,000). Clinico-epidemiological features, based on the 2,523 patients reported from the second survey, were subsequently clarified. The sex ratio (male/female) of the patients was 3.5. Alcoholic pancreatitis was the most common type in males (68.5%), and idiopathic pancreatitis in females (69.6%). Compared with the findings in the last survey in 1985, the proportion of patients with alcoholic pancreatitis has decreased slightly, from 58.7% to 55.5%, while that of idiopathic chronic pancreatitis has increased in both males and females. Patients diagnosed by advanced techniques such as computed tomography (CT) and endoscopic retrograde cholangiopancreatography (ERCP) accounted for 68.1% of the total. The number of patients with chronic pancreatitis treated in 1994 in Japan, was estimated as 32,000, with an overall prevalence rate of 45.4 per 100,000 population in males and 12.4 per 100,000 population in females. PMID- 10680670 TI - Nuclear cyclin D1 overexpression is a critical event associated with cell proliferation and invasive growth in gallbladder carcinogenesis. AB - Cyclin D1 overexpression is remarkably frequent in several human carcinomas and is believed to be a critical event in oncogenesis. We examined cyclin D1 expression, p53 expression, and the Ki-67 labeling index by immunostaining in human gallbladder mucosa in conditions varying from normal to malignant tissue. We also examined K-ras codon 12 mutations in these tissues with a two-step polymerase chain reaction. Nuclear cyclin D1 overexpression was observed in 48% of carcinomas occurring independently of adenoma, but not in adenomas, carcinomas arising in adenomas, or nonneoplastic lesions. Cytoplasmic cyclin D1 overexpression was observed in about 15% of abnormal specimens, irrespective of the type of epithelial abnormality. Carcinomas showing nuclear cyclin D1 overexpression had significantly higher Ki-67 labeling indexes than those with no overexpression. Moderately to poorly differentiated adenocarcinomas showed a higher incidence of nuclear cyclin D1 overexpression than papillary to well differentiated carcinomas. Specimens with cyclin D1 overexpression showed a high incidence of lymph permeation, venous permeation, and lymph node metastasis. We conclude that nuclear cyclin D1 overexpression is a critical event importantly associated with cell proliferation and invasive growth in gallbladder carcinogenesis, and that cyclin D1 immunostaining may become a useful marker for evaluating gallbladder carcinomas. PMID- 10680671 TI - Four resections for hepatic metastasis from gastric cancer: histochemical analysis of cell proliferation, apoptosis, and angiogenesis. AB - In a patient with gastric cancer (GC) associated with one synchronous and three metachronous hepatic metastases (HM), who underwent four hepatectomies, we carried out histochemical investigations regarding cell proliferation, apoptosis, and angiogenesis in the GC and HM. Tissue samples were taken from the primary GC and four HM. Ki-67 immunostaining was performed to evaluate cell proliferation and determine the labeling index (Ki-67 LI; ie, the percentage of cancer cells with nuclei stained for Ki-67). Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) was performed to evaluate apoptosis and determine the apoptotic index (ie, the percentage of TUNEL positive cells), and immunostaining for factor VIII-related antigen was performed to evaluate angiogenesis and measure microvessel density (MVD). The Ki-67 LI was 43.2% in the primary GC and 39.9% in the synchronous HM, and the LI increased with the number of resections of metachronous HM. The apoptotic index was 3.36% in the primary GC, and 5.30% in the synchronous HM, and the index decreased after further resections of the metachronous HM. The MVD was 35 in the primary GC, and 22 in the synchronous HM, and it increased with the number of resections of metachronous HM. The primary GC in this patient may have strongly influenced the growth of HM through effects on cell proliferation, apoptosis, and angiogenesis. PMID- 10680672 TI - Brunner's gland adenoma with a focus of p53-positive atypical glands. AB - A submucosal tumor was resected endoscopically from the duodenal bulb in a 43 year-old man complaining of epigastric discomfort. The tumor, measuring 22 x 20 x 19mm, consisted mainly of Brunner's glands with no atypia. However, close histologic examination disclosed a focus of glands with cellular and structural atypia. The atypical glands showed staining by periodic acid-Schiff, alcian blue, and high iron-diamine methods. Mucin histochemistry was examined, and the atypical glands resembled the excretory ducts rather than the acinar cells of the tumor. Immunohistochemically, positivity for MIB-1 was high (38.0%), and p53 positive cells were detected sporadically in the atypical glands. These results indicated that the atypical glands probably represented a neoplastic lesion. Brunner's gland adenomas associated with foci of true neoplasm are very rare; only two cases, including one patient with microcarcinoid tumors, have been reported. PMID- 10680673 TI - Multiple duodeno-jejunal diverticula causing massive intestinal bleeding. AB - A case of massive intestinal blood loss from multiple duodeno-jejunal diverticula is described. A 39-year-old man was referred to our hospital because of recurrent bloody stool and worsening anemia. Upper and lower endoscopy, selective abdominal angiography, and radionuclide scanning were performed to seek the cause of the intestinal bleeding, but none of these studies revealed the source of bleeding. Small-bowel barium follow-through examination showed numerous diverticula in the distal duodenum and proximal jejunum. Excision of the duodenal diverticulum and resection of the involved portion of the jejunum cured the patient. On histopathological examination, an ulcerative lesion with an exposed vessel suggestive of the source of bleeding was seen in the resected duodenal diverticulum. Although duodeno-jejunal diverticula are rare, the importance of a careful search for this malformation in a patient with intestinal blood loss is stressed. PMID- 10680674 TI - Acquired ileal diverticulum: an unusual bleeding source. AB - Acquired ileal diverticulum is an uncommon condition and diagnosis is often difficult when bleeding occurs from this source. Here we describe two cases of ileal diverticulum with massive bleeding. Both patients presented with anal bleeding, but upper and lower gastrointestinal endoscopy did not reveal the source. Selective visceral angiography finally detected bleeding lesions in the terminal ileum. Surgical resection was performed in both patients, confirming that the bleeding arose from diverticula less than 1 cm in size. In patients with obscure gastrointestinal bleeding, an ileal diverticulum should be considered, and selective visceral angiography should be performed for precise diagnosis. PMID- 10680675 TI - Ischemic hepatitis induced by mesenteric volvulus in a patient with chronic obstructive lung disease. AB - A 66-year-old man with chronic obstructive lung disease was admitted to our hospital, presenting with mesenteric volvulus and mild liver injury. A superior mesenteric angiogram revealed that the arteries supplying the small intestine were twisted in the arterial phase, while the portal vein was not visualized in the late phase. A celiac angiogram demonstrated that portal blood flow from the splenic venous return was maintained. The patient's symptoms had almost resolved the day after admission, and his serum transaminases level had gradually decreased to normal with conservative therapy. A superior mesenteric angiogram on the 13th hospital day showed a normal arteriogram and the portal vein demonstrated blood flow from the superior mesenteric vein. Liver biopsy revealed hemorrhagic necrosis around the central veins, which was compatible with ischemic hepatitis. Since the patient's O2 saturation level on admission was not low enough to have caused ischemic hepatitis by itself, we suspect that a sudden decrease in portal blood flow was the additional factor that allowed the threshold for the initiation of ischemic liver damage to be reached. PMID- 10680676 TI - Is fish oil (n-3 fatty acids) effective for the maintenance of remission in Crohn's disease? PMID- 10680677 TI - Chronic pancreatitis in Japan: is alcoholic pancreatitis decreasing? PMID- 10680678 TI - In vivo MR micro imaging with conventional radiofrequency coils cooled to 77 degrees K. AB - Cryogenically cooled conventional surface coils are shown to provide significant signal-to-noise ratio (SNR) gains for MR micro imaging of tissue structure in vivo. Measurements are described which employ a simple, all-polyvinyl chloride (PVC) vacuum dewar capable of maintaining a bath of liquid nitrogen around the coil, within 5 mm of the tissue to be imaged. Images acquired in vivo at 64 MHz with a 2-cm diameter copper coil cooled to 77 K demonstrated a gain in SNR of approximately 2.7 +/- 0.3 relative to those obtained with the same coil at room temperature under otherwise identical conditions. This increase is consistent with the reduction in coil resistance and the minor contribution to overall resistance from the imaging object. The performance of the coil is illustrated with images from the human finger and rabbit eye and potential applications are discussed. PMID- 10680679 TI - Undersampled projection-reconstruction imaging for time-resolved contrast enhanced imaging. AB - In time-resolved contrast-enhanced 3D MR angiography, spatial resolution is traded for high temporal resolution. A hybrid method is presented that attempts to reduce this tradeoff in two of the spatial dimensions. It combines an undersampled projection acquisition in two dimensions with variable rate k-space sampling in the third. Spatial resolution in the projection plane is determined by readout resolution and is limited primarily by signal-to-noise ratio. Oversampling the center of k-space combined with temporal k-space interpolation provides time frames with minimal venous contamination. Results demonstrating improved resolution in phantoms and volunteers are presented using angular undersampling factors up to eight with acceptable projection reconstruction artifacts. PMID- 10680680 TI - Cardiac real-time imaging using SENSE. SENSitivity Encoding scheme. AB - Sensitivity encoding is used to improve the performance of real-time MRI. The encoding efficiency of single-shot and segmented echo-planar imaging is tripled by means of a 6-element receiver coil array. The feasibility of this approach is verified for double oblique cardiac real-time imaging of human subjects at rest as well as under physiological stress. Sample images are presented with scan times per image down to 13 msec at a spatial resolution of 4.1 mm, and 27 msec at a resolution of 2.6 mm. Moreover, multiple slice real-time imaging is demonstrated at a rate of 38 double-frames per second. PMID- 10680681 TI - A silent event-related functional MRI technique for brain activation studies without interference of scanner acoustic noise. AB - A new data acquisition method for silent, event-related functional MRI in which scanner acoustic noise does not interfere with brain activation is introduced and evaluated in an auditory tonotopic mapping experiment. This method takes into account the hemodynamic-response characteristics of the brain during activation, associated with both task performance and scanner noise. A data acquisition scheme was designed to collect task-induced brain activation signals without interference of scanner noise on stimulus delivery or on the measured response. The advantages of the technique were demonstrated in a tonotopic mapping experiment of human auditory cortex. Tonotopic maps obtained by the technique in normal subjects showed distinct spatial shifts of the activation foci in the lateral part of Heschl's gyrus with changing stimulus frequency, whereas no systematic shift was shown in a conventional event-related experiment using the same stimulation paradigm. Signal change in the activation foci with the new technique was 54% larger than with the conventional technique, suggesting an increased dynamic range of the signal change associated with task-induced brain activation under silent conditions. PMID- 10680682 TI - Assignment of the water slow-diffusing component in the central nervous system using q-space diffusion MRS: implications for fiber tract imaging. AB - Diffusion-weighted NMR spectroscopy (MRS) was performed on isolated bovine optic nerve and rat brain (in vitro) to characterize the multiexponential water signal decay in diffusion experiments. q-Space analysis of the diffusion data was used to obtain structural information about the investigated neuronal tissues. This analysis provided displacement distribution profiles of the water in the sample. Two diffusing components were identified from these profiles, thus enabling us to obtain the following information about the slow decaying component: 1) displacement of this component is restricted to a diffusing distance of approximately 2 microm; 2) it has a longer T2 than the rapidly diffusing component; and 3) the population fraction of this component depends on the orientation of the nerve fiber. When the diffusion was measured perpendicular to the long axis of the bovine optic nerve, the weighting of this population was 41 +/- 2%, whereas parallel to the long axis of the nerve it was found to be 14 +/- 2%. In the randomly oriented brain tissue, the population of this component was only 7 +/- 3%. These observations led to the conclusion that the slow-decaying component originates mainly from restricted water diffusion in the neuronal fibers. In view of these findings, in vitro and in situ diffusion-weighted images with high b values (with long delta) were acquired to obtain highly detailed images of white matter fiber tracts in the central nervous system. These images provide detailed information on white matter fiber tract location and allow spinal cord maturation to be followed with high accuracy. PMID- 10680683 TI - 3D localized 1H-13C heteronuclear single-quantum coherence correlation spectroscopy in vivo. AB - A method for spatially three-dimensional (3D) localized two-dimensional (2D) 1H 13C correlation spectroscopy, localized HSQC, is proposed. This method has the following special feature in the preparation period. The 180 degrees (13C) and 180 degrees (1H) pulses are separated in time, and the 180 degrees (13C) pulse is applied at 1/4 1JCH) before the 90 degrees (1H) polarization transfer pulse. The preparation (echo) period 2tau can then be set substantially longer than 1/(2 1JCH), so that even in a whole-body system, slice-selective 90 degrees (1H) pulses and gradient pulses can be applied in that period. The localization capabilities of this method were confirmed in a phantom experiment. The 3D localized 2D 1H-13C correlation spectra from a monkey brain in vivo were obtained after [1-13C]glucose injection, and amino acid metabolism was detected; that is, [4-13C]glutamate appeared immediately after the injection, followed by the appearance of [2-13C]glutamate, [3-13C]glutamate, and [4-13C]glutamine. PMID- 10680684 TI - 2D-spatial/2D-spectral spectroscopic imaging of intracerebral gliomas in rat brain. AB - 1H-MR spectroscopy in vivo is often hampered by poor spectral resolution. Spectral overlap can be avoided with two-dimensional spectroscopic techniques. Correlation peak imaging has been implemented to measure unambiguously the distribution of several metabolites in a rat brain glioma model. Acquisition weighted spectroscopic imaging reduced the experimental time and provided excellent spatial localization. The choice of an appropriate spectral acquisition window granted good sensitivity. Spectroscopic images presenting a full two dimensional spectrum in every image pixel were acquired in seven rats at 7 Tesla in 195 min, with a nominal voxel volume of 75 microl. Among other metabolites, the distribution of hypotaurine, phosphoethanolamine, alanine, and even glucose could be visualized both in the C6-glioma and in the unaffected brain. PMID- 10680685 TI - Temperature mapping using the water proton chemical shift: self-referenced method with echo-planar spectroscopic imaging. AB - An echo-planar spectroscopic imaging method of temperature mapping is proposed. This method is sufficiently faster than the so-called 3D magnetic resonance spectroscopic imaging (3D-MRSI) method and does not require image subtractions, unlike the conventional phase mapping method when an internal reference signal is detectable. The water proton chemical shift measured by using the tissue lipid as an internal reference clearly visualized the temperature change in a porcine liver sample in vitro. It was also demonstrated that the internally referenced echo-planar spectroscopic imaging method could markedly reduce a temperature error caused by a simple, translational motion between scans compared with the phase-mapping method. PMID- 10680686 TI - Strong field behavior of the NMR signal from magnetically heterogeneous tissues. AB - A theory for the behavior of the nuclear magnetic resonance (NMR) signal obtained from magnetically heterogeneous tissues is developed for the limit of a strong external magnetic field. If BO is the magnitude of the external magnetic field, it is found that a free-induction signal decays in a time scaling as 1/Bo, a single-spin echo signal decays in a time scaling as 1/Bo(2/3), and a multiple spin echo signal decays in a time scaling as 1/Bo(2). Moreover, it is shown that the form of the signal decay for a multiple-spin echo sequence may deviate significantly from an exponential. Numerical results for a model consisting of randomly distributed magnetic spheres are used to confirm the theory. In addition, good agreement is demonstrated between the theory and experimental measurements obtained with particle suspensions. The validity and application of the theory to biological tissues are discussed. PMID- 10680687 TI - Relaxation induced by ferritin and ferritin-like magnetic particles: the role of proton exchange. AB - Proton T1 and T2 in solutions of ferritin and fercayl (a ferritin-like iron dextran particle) solutions were measured, over a wide range of various parameters (Bo, temperature, interecho-time and pH). The window of the previously referred linear dependence of 1/T2 on the static field was increased, up to 500 MHz, and the independence of T2 on the echo time was confirmed. Correlation times were extracted from T1 nuclear magnetic relaxation dispersion profiles. In the pH range studied, no strong variation of the relaxivities of ferritin solutions was noticed. Fercayl, which, unlike ferritin, remains stable under large pH variations, is characterized by strongly pH-dependent relaxation rates. This feature is interpreted as due to the effect of proton exchange in the water relaxation process. Outer sphere theory, which ignores proton binding, is shown to be unable to describe the relaxation of ferritin and ferritin-like particles solutions, first because it predicts a quadratic rate dependence on Bo, but also because it severely underestimates the relaxation rate. Explaining relaxation induced by ferritin and ferritin-like particle solutions will likely require a model that accounts for proton binding. PMID- 10680688 TI - Method to correlate 1H MRSI and 18FDG-PET. AB - The in vivo neuronal contribution to human cerebral metabolic rate of glucose (CMRglc), measured by 18FDG-PET, is unknown. Examining the effect of 1H MRSI derived N-acetyl aspartate (NAA) concentration on positron emission tomography (PET) measures of metabolic activity might indicate the relationship of CMRglc to neuron density. In a population of 19 demented, cognitively impaired, and control subjects, the Miller-Gartner algorithm was applied to whole-brain PET data to isolate the PET signal originating in cortical gray matter alone (GMPET). An analogous procedure applied to multislice proton MRSI data yielded the N-acetyl aspartate concentration in cortical gray matter (GMNAA). In 18 of 19 subjects, a significant linear regression (P < 0.05) resulted when GMPET was plotted against GMNAA, whereby GMPET was higher for higher GMNAA. This suggests that CMRglc rises linearly with increasing neuron density in gray matter. This method may be used to investigate the relationship of CMRglc to neurons in various conditions. PMID- 10680689 TI - Real-time color flow MRI. AB - A real-time interactive color flow MRI system capable of rapidly visualizing cardiac and vascular flow is described. Interleaved spiral phase contrast datasets are acquired continuously, while real-time gridding and phase differencing is used to compute density and velocity maps. These maps are then displayed using a color overlay similar to what is used by ultrasound. For cardiac applications, 6 independent images/sec are acquired with in-plane resolution of 2.4 mm over a 20 cm field of view (FOV). Sliding window reconstruction achieves display rates up to 18 images/sec. Appropriate tradeoffs are made for other applications. Flow phantom studies indicate this technique accurately measures velocities up to 2 m/sec, and accurately captures real-time velocity waveforms (comparable to continuous wave ultrasound). In vivo studies indicate this technique is useful for imaging cardiac and vascular flow, particularly valvular regurgitation. Arbitrary scan planes can be quickly localized, and flow measured in any direction. PMID- 10680690 TI - Functional magnetic resonance imaging in real time (FIRE): sliding-window correlation analysis and reference-vector optimization. AB - New algorithms for correlation analysis are presented that allow the mapping of brain activity from functional MRI (fMRI) data in real time during the ongoing scan. They combine the computation of the correlation coefficients between measured fMRI time-series data and a reference vector with "detrending", a technique for the suppression of non-stimulus-related signal components, and the "sliding-window technique". Using this technique, which limits the correlation computation to the last N measurement time points, the sensitivity to changes in brain activity is maintained throughout the whole experiment. For increased sensitivity in activation detection a fast and robust optimization of the reference vector is proposed, which takes into account a realistic model of the hemodynamic response function to adapt the parameterized reference vector to the measured data. Based on the described correlation method, real-time fMRI experiments using visual stimulation paradigms have been performed successfully on a clinical MR scanner, which was linked to an external workstation for image analysis. PMID- 10680691 TI - A broadband phased-array system for direct phosphorus and sodium metabolic MRI on a clinical scanner. AB - Despite their proven gains in signal-to-noise ratio and field-of-view for routine clinical MRI, phased-array detection systems are currently unavailable for nuclei other than protons (1H). A broadband phased-array system was designed and built to convert the 1H transmitter signal to the non-1H frequency for excitation and to convert non-1H phased-array MRI signals to the 1H frequency for presentation to the narrowband 1H receivers of a clinical whole-body 1.5 T MRI system. With this system, the scanner operates at the 1H frequency, whereas phased-array MRI occurs at the frequency of the other nucleus. Pulse sequences were developed for direct phased-array sodium (23Na) and phosphorus (31P) MRI of high-energy phosphates using chemical selective imaging, thereby avoiding the complex processing and reconstruction required for phased-array magnetic resonance spectroscopy data. Flexible 4-channel 31P and 23Na phased-arrays were built and the entire system tested in phantom and human studies. The array produced a signal-to-noise ratio improvement of 20% relative to the best-positioned single coil, but gains of 300-400% were realized in many voxels located outside the effective field-of-view of the single coil. Cardiac phosphorus and sodium MRI were obtained in 6-13 min with 16 and 0.5 mL resolution, respectively. Lower resolution human cardiac 23Na MRI were obtained in as little as 4 sec. The system provides a practical approach to realizing the advantages of phased-arrays for nuclei other than 1H, and imaging metabolites directly. PMID- 10680692 TI - Interventional MRI-guided brain biopsies using inductively coupled surface coils. AB - The technical realization of inductively coupled surface coils for interventional MR-guided procedures, and the application to brain biopsies in a 0.2 T magnet is described. The advantages compared to standard coils are discussed, and the results of 26 biopsies on eight different neuropathologic diagnoses from varying locations within the brain are presented. Initial experience shows that inductively coupled coils can offer an increased number of indications for interventional procedures in the brain, easier handling of sterility, and often a better access for the surgeon, compared to the use of standard MR head coils. PMID- 10680693 TI - A transmit-only/receive-only (TORO) RF system for high-field MRI/MRS applications. AB - The design and operation of a detunable shielded hybrid birdcage RF head coil optimized for human brain imaging at 170 MHz is presented. A high duty-cycle and rapid-switching decoupling scheme that allows uniform RF transmission with the head coil and reception with a surface coil within the volume of the head coil is also demonstrated. In addition, the circumscribing hybrid coil can be biased to operate as a conventional transmit/receive head coil. Our RF design allows the use of higher sensitivity surface coils or phased-array coils at very high magnetic fields where body RF resonators are not currently available or whose use is precluded by specific-absorption ratio restrictions. The design also allows the use of receive-only coils within head gradient inserts, which normally do not allow transmission with an RF body resonator at any field strength. PMID- 10680694 TI - Demonstration of a compact compressor for application of metastability-exchange optical pumping of 3He to human lung imaging. AB - Hyperpolarized gas magnetic resonance imaging has recently emerged as a method to image lungs, sinuses, and the brain. The best lung images to date have been produced using hyperpolarized 3He, which is produced by either spin-exchange or metastability-exchange optical pumping. For hyperpolarized gas MRI, the metastable method has demonstrated higher polarization levels and higher polarizing rates, but it requires compression of the hyperpolarized gas. Prior to this work, compression of hyperpolarized gas had only been accomplished using a large, complex and expensive apparatus. Here, human lung ventilation images are presented that were obtained using a compact compressor that is relatively simple and inexpensive. For this test, 1.1 bar-L of 15% hyperpolarized 3He gas was produced at the National Institute of Standards and Technology using a modified commercial diaphragm pump. The hyperpolarized gas was transported to the University of Pennsylvania in a holding field provided by a portable solenoid. PMID- 10680695 TI - In vivo intravoxel incoherent motion measurements in the human placenta using echo-planar imaging at 0.5 T. AB - This paper presents the first in vivo measurements of intravoxel incoherent motion in the human placenta, obtained using the pulsed gradient spin echo (PGSE) sequence. The aims of this study were two-fold. The first was to provide an initial estimate of the values of the IVIM parameters in this organ, which are currently unknown. The second aim was then to use these results to optimize the sequence timings for future studies. The moving blood fraction (f), diffusion coefficient (D), and pseudo-diffusion coefficient (D*) were measured. The average value of f was 26 +/- 6 % (mean +/- SD), D was 1.7 +/- 0.5 x 10(-3) mm2/sec, and D* was 57 +/- 41 x 10(-3) mm2/sec. For the optimized values of b, the expected percentage uncertainty in the fitted values of f, D, and D* for the placenta were sigmaf/f = 14.9%, sigmaD/D = 14.3%, sigmaD*/D* = 44.9%, for an image signal-to noise of 20:1, and a total imaging time of 800 sec. PMID- 10680696 TI - Fast functional MRA using time-resolved projection MR angiography with correlation analysis. AB - Most recently, time-resolved 2D MRA after injection of a contrast agent bolus for various applications has been proposed. Similar to conventional digital subtraction angiography (DSA), 2D MR DSA offers the ability to observe the dilution of the bolus in the vascular system during the passage with a temporal resolution considerably below 1 sec. The purpose of this paper is to present strategies to improve the inherent low signal-to-noise ratio of 2D angiograms while retaining some temporal resolution. This can be achieved by applying algorithms for time series analysis as used in functional MRI. The significantly improved image quality is demonstrated on examples from clinical studies from bronchial MRA as well as cardiovascular MRA. In addition to the increased signal to-noise ratio, correlation analysis leads to suppression of background signals and to a better discrimination of overlapping vessels. Further improvements in the temporal discrimination of vessels is afforded by the use of consecutive multiple contrast agent boli as demonstrated by numerical simulations and experiments. PMID- 10680697 TI - A median filter for 3D FAST spin echo black blood images of cerebral vessels. AB - High-resolution black-blood MRA images of intracranial vascular anatomy can be obtained using 3D fast spin-echo techniques. Although these images demonstrate excellent contrast between vessels and surrounding soft tissues, the dark signal from air and bone can obscure the desired vascular information when a minimum intensity projection image is created. In this paper, we describe an image processing technique based upon a median filter that is effective for detecting narrow vessel-like structures. Minimum intensity projection images of the filtered MRA volume can be obtained in any orientation without prior segmentation of the skull or surrounding air spaces. The filter is very effective for detecting and visualizing small vessels, but is much less effective for detecting vessels and vascular pathology larger than the filter detection width. The filtering technique is demonstrated on black-blood MRA images from a volunteer study. PMID- 10680698 TI - Cardiac-respiratory gating method for magnetic resonance imaging of the heart. AB - In studies of transmural myocardial function, acquisitions of high spatial and temporal resolution tagged cardiac images often exceed the practical time limit for breath-hold fast imaging techniques. Therefore, a dual cardiac-respiratory gating device has been constructed to acquire SPAMM-tagged cardiac MR images at or near end-expiration during spontaneous breathing, by providing an external trigger to a conventional MRI system. Combined cardiac and respiratory gating essentially eliminates the respiratory motion artifacts in tagged cardiac MR images. Compared to cardiac-gated images obtained during intermittent breath holds, cardiac-respiratory gated images show improved tag-myocardium contrast due to magnetization recovery during inspiration. PMID- 10680699 TI - Field mapping without reference scan using asymmetric echo-planar techniques. AB - Improvements in Bo mapping and shimming were achieved by measuring the static field information in multiple subsequent echoes generated by an asymmetric echo planar readout gradient train. With careful compensation, eddy current effects were shown to affect the adjustment of the shim coils minimally. In addition to reducing the time required for field mapping by two-fold, the sensitivity was simultaneously optimized irrespective of the prevalent T2* present, thereby minimizing the error of the static field measurement to below 0.1 Hz. With adiabatic low flip-angle excitation, the time required for field mapping was below 1 second. PMID- 10680700 TI - Expression of complement regulatory proteins CR1, DAF, MCP and CD59 in haematological malignancies. AB - Host cells are protected from the lytic effect of the complement system by complement regulatory proteins. This study was designed to investigate the expression of complement regulatory proteins on leukemic blasts which may be susceptible to the lytic effects of the complement system in the circulation. The surface expressions of complement regulatory proteins, complement receptor 1 (CR1, CD35), decay accelerating factor (DAF, CD55), and homologous restriction factor 20 (HRF20, CD59), on peripheral blood and bone marrow blasts were evaluated by using flow cytometry in 16 acute myeloblastic leukemia (AML), 16 acute lymphoblastic leukemia (ALL), 4 chronic lymphocytic leukemia (CLL), 3 chronic myelocytic leukemia (CML) patients and control granulocytes and lymphocytes obtained from 15 healthy volunteers. mRNA expression was investigated by Northern blot analysis. mRNA abundances were calculated after normalization according to 28s rRNA. Surface expressions of CRI and DAF were marginally (p = 0.08 and p = 0.08, respectively) lower in AML, and DAF expression was significantly lower (p=0.0008) in ALL patients in comparison to their normal counterparts. Except from a slight increase that is detected for CD59 in CML patients (p=0.06), there was no significant difference between the surface expressions of CD59 in any of the groups studied. Densitometric analysis of autoradiographs obtained from Northern blots revealed that in AML patients, CR1 mRNA expression were 5.5-fold lower than controls (p=0.06), while DAF mRNA expression was significantly higher (p=0.0046). Furthermore, the mRNA expression of CRI in ALL patients was found significantly lower than in the control group (p = 0.0419). None of the values obtained from the other groups were significantly different from each other. These results suggest that leukemic blasts are protected from the lytic attack of the complement system at all levels, since all of the complement membrane regulatory proteins were expressed in all leukemia types (although at lower amounts in some cases), and it is also possible to use CRI and DAF as differentiation markers in acute leukemias. PMID- 10680701 TI - Cell kinetic study of normal human bone marrow hematopoiesis and acute leukemia using 7AAD/PY. AB - We have used the 7AAD/PY method to analyze the cell cycle status of normal human bone marrow hematopoiesis, and found that the cell kinetics differed. There were cells with relatively low levels of RNA in the S-phase (Type I) and a high level in the S-phase (Type 11). T-cells, B-cells, nucleated red cells and CD34+/CD19+ early B-cells in bone marrow were Type I, whereas myelomonocytic subset and CD34+/CD33-dim+ common myeloid cells were Type II. AC133+/CD38-dim cells, which were thought to be lineage-marker negative hematopoietic stem cells, had intermediate amounts of RNA in the S-phase between Type I and II (Type 0). Seventy-four cases of acute leukemia were also analyzed. Most of the T- and B-ALL cases were found to be Type I, most of the ANLL cases were Type II, and there were 10 cases that were Type 0. These findings yielded fundamental information about normal hematopoiesis and acute leukemia. PMID- 10680702 TI - Increased liposome-mediated gene transfer into haematopoietic cells grown in adhesion to stromal or fibroblast cell line monolayers. AB - We investigated transfection rates of CD34+ haematopoietic progenitor cells (HPC) or haematopoietic cell lines (TF-1, KG1a and K562) using the LacZ gene as a reporter and cationic liposomes. The transfection efficiency of CD34+ haematopoietic progenitor cells (HPC) or TF-1, KG1a and K562 grown in suspension is very low (average percentage of 0.013 for HPC and 0.03 for cell lines). Adhesion of HPC or cell lines to plates by immunological or physical methods significantly enhances transfection efficiency; however, the percentage of transfected cells still remained low. We found that adhesion of TF-1, KG1a and K562 HC to MS-5 stroma cells or NIH-3T3 fibroblast cells increased transfection efficiency. Under these conditions transfection is achieved in 11.2-25% (mean 18.30%) for the cell lines and 13.6% (range 8.2-24.2%) for CD34+ HPC. These results indicate that liposome-mediated transfection of HC is significantly increased when cells are grown in adherence to stroma or fibroblast monolayers. PMID- 10680703 TI - Polycythaemia vera: bone marrow histopathology under treatment with interferon, hydroxyurea and busulphan. AB - Little is known about long-term effects of myelosuppressive therapy on bone marrow of patients with polycythaemia vera, since histopathology from follow-up biopsies has not been frequently reported. Thus we conducted a retrospective morphometrical analysis of diagnostic and follow-up biopsies of 62 patients, evaluating fibre content, megakaryocytes and bone marrow cellularity. 8/62 patients were treated with interferon-alpha (INF), 11/62 with hydroxyurea (HU) and 11/62 with busulphan (BU). 32/62 served as controls; they were not treated with myelosuppressive drugs but with phlebotomy only. The median observation time was 2.3 yr. Results were compared on the basis of change per time. The bone marrow of the patients with phlebotomies only was characterised by increasing cellularity of haematopoesis, number and volume ratio of megakaryocytes and fibre content. In BU- and HU-treated patients, the haematopoesis was significantly reduced. The IFN patients revealed a reduction of cellularity which was not significant. The fibre content was reduced by BU only, but not significantly. No correlation between megakaryocytes and fibres was found. It could be concluded therefore that: 1) fibre proliferation within the bone marrow was not significantly altered by IFN, HU or BU. 2) Cellularity of haematopoesis was reduced significantly by HU and BU but only partly by IFN, corresponding with haematological remission. PMID- 10680704 TI - Neonatal and maternal thrombocytopenia: incidence and immune background. AB - We performed a prospective study on the incidence of thrombocytopenia (t-penia) and its immunological origin in unselected 26,275 mothers and 24,101 newborns. Platelet antibodies were examined by the platelet immunofluorescence test (PIFT) and the monoclonal antibody immobilisation of platelet antigens assay (MAIPA). T penia (platelet count < 100 x 10(9)/I) was found in 124 (0.5%) mothers (in 0.04%) severe, <50x 10(9)/l) and in 116 (0.5%) newborns (in 0.15% severe); 90 (72.6%) and 112 (96.6%), respectively, were available for further studies. In both groups non-immune t-penia was diagnosed about 4.5 times more often than the immune t penia. Among 90 mothers, t-penia was severe in 11.1%, antibodies were detected in 17.8%; both factors were not prognostic for delivering thrombocytopenic newborns. Among 112 babies, 21 were delivered by thrombocytopenic mothers and 91 by mothers with normal platelet count; among newborns with immune t-penia the proportion of alloimmune (NAIT) to autoimmune was equal (10 with NAIT, 10 with autoimmune, 4 of them born by mothers with hidden autoimmune t-penia). In 33% of the neonates t penia was severe, most often among NAIT. In conclusion, although t-penia in mothers as well as in infants is not frequent and severe, and an immune origin not often found, the search for antibodies, in particular alloantibodies, should be done. Even if the serological results are not helpful at the moment, they can be of importance in subsequent pregnancy and for related pregnant women. PMID- 10680705 TI - Alpha-interferon as induction and maintenance therapy in hairy cell leukemia: a long-term follow-up analysis. AB - Although in recent years the use of purine analogues has increased the percentage of long-term complete response the effect on overall survival of patients with hairy cell leukemia (HCL) is not yet clear. This study aimed to evaluate the long term outcome (mean follow up of 92 months) of 64 patients receiving IFN as first line therapy. IFN was well tolerated and effective. The overall response rate was 91% (PR 65%, CR 13%, GPR 13%). Forty-one patients (63%) received IFN 3 MU/ wk as maintenance therapy. The 10-yr projected survival rate of responding patients (CR and GPR 100%; PR 95%) and non-responders (SD, PD 80%) clearly shows that type of response does not affect survival. Patients receiving IFN maintenance had a statistically higher PFS than those who did not (p <0.01). This study shows that IFN is still one of the standard therapies for this disease, that achieving CR has no primary relevance for the control of the disease, and that good utilization of therapeutic resources may assure HCL patients a survival rate comparable to that of a normal, healthy population. PMID- 10680706 TI - Association of the alpha-spectrin R28H mutation with allele alphaLELY and with alphaI/alphaII domain haplotypes in three Brazilian families. AB - We have studied three Brazilian kindreds presenting spectrin alpha/74 hereditary elliptocytosis (HE) due to a G-->A substitution, responsible for the R28H mutation. The mutant allele was associated with alphaI domain haplotype 1 (XbaI /MspI-/PvuII+) in all three families and with two different alphaII domain haplotypes (1/RIT, 4/RVR). This result may reflect that this mutation occurs in a "hot spot" and may have arisen more than once or that a crossing over event may have occurred between the two domains studied. We detected one new haplotype in the alphaI domain (haplotype 3 -XbaI(+)/MspI(-)/PvuII(+)). The mutant allele was associated with the lack of the alphaII domain Alu insertion in all three cases. Allele alphaLELY, detected by PCR and restriction enzyme digestion, was present in the heterozygous form in patient 1 (alphaHE/alphaLELY) and in the homozygous form in patients 2 and 3(alphaHE-LEL/alphaLELY). It was found to be associated with domain haplotypes I (RIT) and 4 (RVR) and with the presence and absence of the Alu insertion. This may have arisen through recombination events, since this polymorphism is located in the alphaIV-alphaV domain junction, which is far distant from the alphaII domain. PMID- 10680707 TI - Hereditary haemorrhagic telangiectasia with protein S deficiency in a family: a case report. PMID- 10680708 TI - Effect of biscoclaurine alkaloids, prednisolone and ascorbic acid on myelodysplastic syndrome with pancytopenia: a case report. PMID- 10680709 TI - CD45-negative acute leukemia in adulthood. PMID- 10680710 TI - Gammadelta T cell lymphoproliferation: a case report. PMID- 10680711 TI - Relative and combined effects of ethanol and protein deficiency on bone histology and mineral metabolism. AB - This study was performed to analyze the relative and combined effects of ethanol and protein deficiency on bone histology and mineral metabolism in 4 groups of 7 animals each which were pair-fed during 8 weeks with 1) a nutritionally adequate diet; 2) a 36% (as energy) ethanol containing isocaloric diet; 3) a 2% protein, isocaloric diet; and 4) a 36% ethanol 2% protein isocaloric diet, respectively, following the Lieber-DeCarli model. Another group of five rats were fed ad libitum the control diet. The first and second lumbar vertebrae were removed after sacrifice, and processed for histomorphometrical analysis of undecalcified bone samples. Blood and 24-h urine were also collected. Protein malnutrition, but not ethanol, leads to osteoporosis and reduced osteoid synthesis, whereas ethanol and protein malnutrition both lead to impaired bone mineral apposition and increased urinary hydroxyproline excretion. These changes are accompanied by an increase in serum parathormone and serum 1,25 dihydroxy vitamin D3, a slight hypomagnesemia, hypercalciuria and hyperphosphaturia; protein deficiency plays an independent role in these alterations, whereas both ethanol and protein deficiency exert independent effects on decreasing serum testosterone levels; this last alteration may contribute to the bone changes mentioned before. PMID- 10680712 TI - Influence of ethanol on lead distribution and biochemical changes in rats exposed to lead. AB - In the present study, an attempt has been made to investigate the effect of ethanol consumption on the distribution of lead in different regions of brain and body organs of male albino rats. Lead when administered intragastrically, for a period of eight weeks resulted in almost uniform accumulation of this metal in all the regions of brain, which increased by almost two fold when ethanol was given along with lead. Lead was also seen to compartmentalise in almost all the tissues of the body to varying extents, with the highest accumulation in the kidney. A progressive and appreciable accumulation of lead was seen in blood with a concomitant increase in ZPP levels in animals during the course of treatment, which increased further when ethanol was administered along with lead. The activity of delta-ALAD and AChE in blood was significantly decreased in lead as well as ethanol treated animals. However, in animals coexposed to lead and ethanol, the inhibition of delta-ALAD was not significantly different, when compared to only lead-treated animals. The results suggested that animals exposed to ethanol and lead simultaneously accumulate higher levels of lead in blood and brain of animals making them more vulnerable to the haematological and neurological toxic effects of lead. PMID- 10680713 TI - Effects of phentermine and fenfluramine on alcohol consumption and alcohol withdrawal seizures in rats. AB - The drug combination of phentermine plus fenfluramine has been used clinically in both the treatment of obesity and alcoholism. The aim of the current study was to assess the interaction of the two drugs on consumption of both an alcohol containing and a nonalcoholic diet. Furthermore, the efficacy of the drug combination on suppression of withdrawal seizures was determined. Animals were either maintained on a 6% alcohol-containing diet, free-fed an isocaloric control, or pair-fed the control diet. It was observed that, with regard to body weight growth curves, alcohol provides about 2.5 kcal/g. Both phentermine and fenfluramine caused a decrease in consumption 1 h after administration; however, during the next 23 h, 4 mg/kg phentermine significantly increased consumption of all diets. At doses of 1 and 2 mg/kg, fenfluramine selectively reduced consumption of the alcohol-containing diet as compared to the isocaloric diets. Lower doses of fenfluramine blocked the increases in consumption induced by phentermine. Furthermore, in animals fed the nonalcoholic diet, the drug combination of 2 mg/kg fenfluramine plus 8 mg/kg phentermine produced a 63-82% reduction in consumption, an effect not seen when either drug was administered alone. This greater than additive effect was also seen in the earlier time periods in animals pair-fed the control diet. Neurochemical analysis from these animals revealed that the alcohol-dependent animals displayed a significant reduction of DOPAC and 5-HIAA levels in the striatum, frontal cortex, and hypothalamus after a 9-h withdrawal period, further implicating the serotonergic and dopaminergic systems in mediation of withdrawal symptoms and alcohol craving. Finally, 8 mg/kg phentermine plus 8 mg/kg fenfluramine completely abolished alcohol withdrawal seizures, compared to a 78% rate in saline treated rats. In conclusion, the coadministration of phentermine plus fenfluramine produced a moderate reduction of alcohol consumption and was completely effective at reducing alcohol withdrawal seizures. PMID- 10680714 TI - Motor impairment produced by ethanol and site-selective NMDA receptor antagonists in mice: tolerance and cross-tolerance. AB - Current perspectives on clinical use of N-methyl-D-aspartate (NMDA) receptor antagonists infer acute and repeated administration schedules for management of different pathological states. Development of tolerance and cross-tolerance between different antagonists may significantly affect their clinical effectiveness. Since ethanol was repeatedly demonstrated to act as NMDA receptor antagonist, ethanol use may also have its impact on the effects of NMDA receptor ligands. Using the rotarod test in mice, the present study evaluated development of tolerance and cross-tolerance between ethanol (3.2 g/kg, p.o.), competitive NMDA receptor antagonist, D-CPPene (5.6 mg/kg, i.p.), and low-affinity NMDA receptor channel blocker, memantine (30 mg/kg, i.p.), that were administered for seven days once a day after the daily rotarod training session. Acute tests with ethanol (0.3, 1, 1.7, 3.2 g/kg), D-CPPene (0.3, 1, 3, 5.6 mg/kg) and memantine (1, 3, 10, 30 mg/kg) revealed that (a) each of these drugs dose-dependently disrupted rotarod performance in drug-naive mice; (b) in ethanol- and D-CPPene treated mice, tolerance was observed to ethanol and D-CPPene but not to memantine; moreover, effects of memantine were even more pronounced in D-CPPene treated subjects; and (c) repeated memantine administration decreased acute motor impairing effects of ethanol, D-CPPene and memantine. Thus, the history of ethanol use or abuse may influence pharmacological activity of NMDA receptor antagonists and this effect is dependent on type of the NMDA receptor antagonist applied. PMID- 10680715 TI - Lipopolysaccharide-stimulated nitric oxide production and inhibition of cell proliferation is antagonized by ethanol in a clonal macrophage cell line. AB - Both chronic and acute ethanol exposure have been shown to be cytotoxic and also to disrupt normal cell function or responses in a variety of cell types. Macrophage function has specifically been shown to be disrupted by chronic ethanol exposure by mechanisms that have not been elucidated. It is known that exposure of macrophages to lipopolysaccharide (LPS) from gram-negative bacteria will decrease the number of cells. Since increased exposure to endotoxin is often associated with chronic alcoholism, this may be one mechanism to account for loss of macrophages in alcoholic patients. The loss of macrophages, as a consequence of endotoxin treatment, appears to be linked to cell activation and, in particular, LPS-stimulated synthesis of nitric oxide which has been suggested to cause an increase in apoptosis. Ethanol also increases apoptosis in some cell types but, in general, ethanol inhibits activation of macrophages. Thus, the overall effect on cell numbers and cell proliferation elicited by treating macrophages concomitantly with ethanol and LPS depends on the balance between inhibiting LPS-mediated activation and the actions of ethanol. The interaction between ethanol and LPS was investigated in a macrophage cell line (RAW 264.7 cells) by measuring nitric oxide production and cell proliferation. A 24-h exposure to ethanol (100 mM) decreased [3H]-thymidine incorporation significantly. LPS treatment elicited a concentration-dependent decrease in [3H] thymidine incorporation at LPS concentrations of 0.1 ng/ml to 1000 ng/ml and stimulated nitric oxide production at concentrations above 1 ng/ml. LPS stimulated nitric oxide production was inhibited by ethanol (20 to 100 mM) and the nitric oxide synthesis inhibitor, N(G)Nitro-L-arginine methyl L-NAME) ester (100 and 500 microM). However, LPS-inhibited [3H]-thymidine incorporation was not be totally reversed by ethanol- or L-NAME-treatment. A direct correlation between nitric oxide production and inhibition of cell proliferation could not be demonstrated. However, it appears that ethanol and LPS do affect some common mechanism(s) in this cell line. PMID- 10680716 TI - Ontogeny of ethanol elimination and ethanol-induced hypothermia. AB - Ontogeny of ethanol elimination rates and ethanol-induced hypothermia were examined as possible mechanisms contributing to the marked reduction in ethanol sensitivity early in life (Little et al., 1996; Silveri & Spear, 1998) and the notable gender difference in ethanol sleep-time seen in adult animals (Silveri & Spear, 1998). Elimination rates and brain/blood ethanol levels were determined following doses of 1.5 or 4.5 g/kg ethanol in male and female Sprague-Dawley rats at postnatal days (P)16, 26, 36, or 56. Animals were sacrificed at 40, 80, or 160 min post-injection, with ethanol elimination rates estimated from the slope of the regression of blood and brain alcohol levels across the three sampling periods. P16 animals exhibited the slowest rate of ethanol metabolism, while no gender effects were evident at any age. Observed ontogenetic increases in ethanol hypothermia were not systematically related to the ontogeny of ethanol metabolism. Factors other than ontogenetic changes in ethanol metabolism, hypothermia, or the distribution of ethanol between brain and blood must underlie the relative insensitivity to ethanol often reported in young and adolescent organisms, a fruitful area for future studies given the frequent use and misuse of alcohol by human adolescents. PMID- 10680717 TI - Reaction of nitroxyl, an aldehyde dehydrogenase inhibitor, with N-acetyl-L cysteine. AB - Nitroxyl (HNO) is the aldehyde dehydrogenase (AIDH) inhibitor produced by catalase action on cyanamide. Incubation of N-acetyl-L-cysteine (NAC), a reagent with a free sulfhydryl group, with Piloty's acid (a nitroxyl generator) suggested that NAC was acting as a competitive "trap" for nitroxyl. Elucidation of the structure of this reaction product should give an insight as to how nitroxyl interacts with AIDH, a sulfhydryl enzyme. We now present evidence that the product formed is N-acetyl-L-cysteinesulfinamide (NACS). We have synthesized NACS and showed that this synthetic product was identical to the product formed in the trapping experiment. Both had identical RT values by reverse phase HPLC and identical RF values by TLC using three different solvent systems. The structural identification of this nitroxyl trapped product as a sulfinamide now allows the chemical confirmation of the active-site cysteine residue of AIDH as Cys-302. PMID- 10680718 TI - Haloperidol administered subchronically reduces the alcohol-deprivation effect in mice. AB - During the pre-experimental phase, hybrid (CBA x C57BL) male mice having had 16 weeks free access to food, water and flavored 30% alcohol were deprived of alcohol for 3 days. The next day they were given free choice between similarly flavored water and 30% alcohol. The mice were divided into two subgroups having (HD) or lacking (LD) the deprivation-induced elevation in alcohol intake during the first 1.5 h of renewed access compared with their intake during the last 22.5 h of first postdeprivation day. In Experiment 1, alcohol naive, LD, and HD mice received daily injections of haloperidol (Haldol; 1 mg/kg) or vehicle during 14 days of abstinence. The behavior of the mice was evaluated in an exploratory cross-maze and inescapable slip funnel test a day after the 13th injection (before the 14th injection). On the first postinjection day, the mice were again given a free choice between flavored water and alcohol. In Experiment 2, all the mice were administered with vehicle during the first 13 days of abstinence. On 14th day, they received an injection of haloperidol (1 mg/kg) or vehicle and a day later were given choice between flavored water and alcohol. Unlike a single injection, the subchronic administration of haloperidol lowered the alcohol intake by HD mice with a more prominent decrease seen during the first 1.5 h than during the last 22.5 h of first postdeprivation day. The alcohol-deprivation effect in HD mice decreased by 79% after subchronic haloperidol. No significant change in alcohol intake was found in alcohol-naive and LD mice. Water intake did not vary systematically. Among the groups, the effect of subchronic haloperidol on the alcohol-deprivation effect did not parallel changes in most of the measures of exploratory or avoidance behavior. It is proposed that haloperidol administered subchronically may attenuate motivation for alcohol. PMID- 10680719 TI - Anxiety in mice following acute aspartame and ethanol exposure. AB - The purpose of the present study was to look at the effect of aspartame on the anxiolytic actions of ethanol. Previous research has shown that ethanol reliably produces an anxiolytic effect on rodent's plus-maze performance. There have been anecdotal reports that aspartame increases anxiety. CD-1 male mice were given i.p. aspartame doses of vehicle, 1000, or 2000 mg/kg, followed 30 min later by i.p. ethanol doses of 1.6 g/kg or vehicle. Animals were then placed in an open field, then tested in the plus-maze. Results determined that the aspartame condition had no significant effect on anxiety-related behavior, nor did it alter the anxiolytic actions of ethanol. Thus, acute high dose exposure to aspartame does not appear to affect anxiety-related behaviors. PMID- 10680720 TI - Differential sensitivity of mouse neural crest cells to ethanol-induced toxicity. AB - Neural crest cells (NCCs) have been identified as an important target population relative to ethanol-induced teratogenicity in both mouse and avian models. Additionally, whole embryo culture mouse models have shown strain-related differences in sensitivity to ethanol-induced damage following acute exposure during early NCC development. That differential sensitivity of NCCs may contribute to these strain differences has been unexplored. For this purpose, cultured NCCs from an inbred mouse strain (C57BL/6J; C57) that is more sensitive to ethanol-induced teratogenicity than an outbred strain (ICR) were compared. This study showed that the incidence of cell death was significantly higher for the C57 NCCs than those from the ICR strain at all ethanol concentrations tested, and as early as 12 hours after initial exposure to 100 mM ethanol. The lateral mobility of the membrane lipids was faster and the membrane GM1 content was lower in C57 cells than ICR cells both under control conditions and at all doses and times tested. Ethanol exposure resulted in significant increases in the membrane lipid lateral mobility, and decreases in the membrane GM1 content that occurred in a dose and time-dependent manner in the NCCs from both strains. A significant correlation was found between the GM1 content and lateral mobility of the membrane lipids, the lateral mobility of membrane lipids and cell viability, as well as the GM1 content and cell viability in the NCCs from both strains. These results suggest that different strain sensitivities to ethanol-induced teratogencity may lie, at least in part, in the interstrain differential response of the NCC population and that the vulnerability of the NCCs to ethanol-induced death may be related to their endogenous membrane GM1 content. PMID- 10680721 TI - The role of dietary fat in alcohol's prenatal effects. AB - Pregnant rats were fed a control diet or high saturated fat diet (lard) for 6 weeks prior to breeding and continued to consume these diets during pregnancy. Beginning on gestation day 8, rats in each diet group were intubated with 5.5 or 0 g/kg alcohol. Rats in the 0 g/kg group were pair-fed to those in their respective 5.5 g/kg groups. Offspring were weighed at birth. On postnatal days 18 and 20, they were tested for passive avoidance learning and locomotor activity, respectively. Animals prenatally exposed to alcohol weighed less at birth and at weaning time (21 days of age) and required more trials to reach criterion in the passive avoidance test but did not differ in activity. Diet did not affect any of these measures significantly nor were there any significant interactions. We conclude that high saturated fat from lard does not influence alcohol's prenatal effects. PMID- 10680722 TI - Thyrotropin releasing hormone decreases alcohol intake and preference in rats. AB - Thyrotropin releasing hormone (TRH) has been reported to reduce stress- and deprivation-induced eating, hypothetically by induction of satiation. Early work demonstrated thyroid extracts reduced alcohol intake, and recent research shows a TRH analog specifically inhibits alcohol preference. We determined whether parenteral administration of TRH reduces alcohol consumption and choice in a manner consistent with a satiation effect. Water-restricted ad lib fed female and male rats (n = 12) were given access to 5% w/v ethanol 0 or 30 minutes after intraperitoneal (i.p.) injection of TRH. TRH (20-40 mg/kg) inhibited alcohol intake only if injected immediately before alcohol access. Inhibition of alcohol intake was reliably accompanied by increased production of fecal boli but not by reliably decreased food intake. Rats given a choice of 2% w/v ethanol and water decreased alcohol preference after TRH (20 mg/kg) but did not reduce total fluid intake. Results are partially consistent with the hypothesis of TRH as one of several functional elements in the integrative neuropeptide control of alcohol consumption via short-term satiation. PMID- 10680723 TI - Neonatal alcohol exposure produces more severe motor coordination deficits in high alcohol sensitive rats compared to low alcohol sensitive rats. AB - Prenatal exposure to alcohol can produce a number of behavioral alterations, including hyperactivity, learning deficits and motor impairments. However, the severity and nature of behavioral alterations varies markedly among children of women who drink during pregnancy. One important determinant of this variation may be genetic differences in the response to alcohol. Recently, we demonstrated that exposure to alcohol during development produced hyperactivity in rats bred for high alcohol sensitivity (HAS), but not in rats bred for low alcohol sensitivity (LAS). These lines were selectively bred for extremes in alcohol-induced "sleep time." The present study investigated the effects of ethanol exposure during development on motor coordination later in life in both HAS and LAS rats. Using an artificial rearing procedure, neonatal pups from each line were exposed to a binge-like alcohol treatment on postnatal days (PD) 4-9. Within each line, one group was exposed to ethanol (6.0 g/kg/day), one group served as an artificially reared control, and a third served as a normally reared control group. On PD 30, parallel bar motor performance was evaluated. Exposure to ethanol during development severely impaired motor performance in the HAS rats compared to their controls. In LAS rats, early ethanol exposure produced only mild and nonsignificant effects on motor performance. Thus, HAS rats were more vulnerable to ethanol-induced motor deficits compared to the LAS rats. Importantly, there were no differences in peak blood alcohol level between the lines, indicating that vulnerability to ethanol's teratogenic effects was not due to differences in metabolic rate. These results suggest that genetic differences in response to alcohol may serve as a predictor for susceptibility to ethanol's teratogenic effects. PMID- 10680724 TI - Western diet, family history of colorectal cancer, NAT2, GSTM-1 and risk of colon cancer. AB - OBJECTIVE: In this study we examine the combined effects of Western diet, age at diagnosis, and genetic susceptibility. METHODS: We use data collected as part of an incident case-control study of colon cancer. Family history of colorectal cancer, N-acetyltransferase (NAT2), and glutathione-S-transferase (GSTM-1) are studied with Western diet and age at diagnosis. RESULTS: A significant interaction between age at time of diagnosis, Western dietary pattern, and family history of colorectal cancer (p for interaction = 0.03) was detected. Those with a family history of colorectal cancer who ate a predominantly Western diet were at increased risk of colon cancer (OR 14.0, 95% CI 3.9-50.1 for < or = 55 years; OR 7.7, 95% CI 2.0-29.1 for 56-66 years; OR 1.6, 95% CI 0.8-3.2 for > or = 67 years) compared to those without a family history of colorectal cancer and low levels of a Western diet. Associations with the Western diet were stronger than individual components of the dietary pattern. Neither NAT2 nor GSTM-1 showed significant interaction with Western diet. CONCLUSION: The extent to which diet comprising a Western dietary pattern influences risk of colon cancer is dependent on age. This dietary pattern also appears to modulate the colon cancer risk associated with a family history of colon cancer. PMID- 10680725 TI - World Cancer Research Fund/American Institute of Cancer Research 1997 recommendations: applicability to digestive tract cancer in Japan. AB - OBJECTIVES: This paper reviewed analytic epidemiological studies of the major Japanese digestive tract cancers, i.e. esophageal, stomach, colon and rectal. The applicability of the recommendations for prevention of these cancers by the World Cancer Research Fund/American Institute of Cancer Research (W&A) to Japan is considered. METHODS: Papers were searched by the MEDLINE for the period 1966 through 1997. Among them, 43 relevant papers including data from Japan were reviewed. RESULTS: Results for 11 lifestyle-related factors were considered. Cigarette smoking was a strong and consistent, thus, convincing, risk factor for esophageal cancer, and a possible risk factor for stomach and colorectal cancer. Excessive consumption of alcohol was a convincing risk factor for esophageal cancer, and a possible risk factor for stomach and colorectal cancer. Excessive salt intake was a risk factor supported by some strong evidence but inconsistent; therefore, it is a probable risk factor for stomach cancer and a possible risk factor for colorectal cancer. Low physical activity was a probable risk factor for colorectal cancer. On the other hand, sufficient intake of vegetables, including green-yellow vegetables, and fruits was regarded as a possible protective factor for these cancers. CONCLUSIONS: These observations were mostly consistent with those reported by W&A; therefore the recommendations by W&A for prevention of these cancers may be considered applicable to the current Japanese population. PMID- 10680726 TI - Vitamin D receptor genotype and breast cancer in Latinas (United States). AB - OBJECTIVE: Polymorphism in the vitamin D receptor (VDR) gene has been associated with variation in bone mineral density and with prostate cancer risk. The purpose of this study was to determine whether polymorphism in the VDR gene may also influence breast cancer risk. METHODS: Polymorphisms in the 5' and 3' ends of the VDR gene were genotyped for 143 Latina women with breast cancer and 300 cohort controls. RESULTS: Both the BsmI and poly-A polymorphisms in the 3' end of the VDR gene were associated with breast cancer risk, with a trend for increasing risk with increasing number of BsmI B alleles or short (S) poly-A alleles. Compared to subjects having two long poly-A alleles (genotype LL), odds ratios (and 95% confidence intervals) were 1.5 (1.0 2.3) and 3.2 (1.5-6.9) for subjects having genotypes SL and SS, respectively. Compared to BsmI genotype bb, odds ratios (and 95% confidence intervals) were 1.6 (1.1-2.5) and 2.2 (1.0-4.7) for genotypes Bb and BB respectively. The start codon polymorphism, FokI, was not associated with breast cancer risk. CONCLUSION: These results suggest that polymorphic variation in or near the 3' end of the VDR gene influences breast cancer risk in Latina women. PMID- 10680727 TI - Partitioning linear trends in age-adjusted rates. AB - OBJECTIVE: Surveillance of chronic diseases includes monitoring trends in age adjusted rates in the general population. Statistics that are calculated to describe and compare trends include the annual percent change and the percent change for a specified time period. However, it is also of interest to determine the contribution specific diseases make to an overall trend in order to better understand the impact of interventions and changes in the prevalence of risk factors. The objective here is to provide a method for partitioning a linear trend in age-adjusted rates into disease-specific components. METHODS: The method presented is based on linear regression. The decreasing trend in age-adjusted cancer mortality rates for the total United States during the period 1991-96 is analyzed to illustrate the method. RESULTS: Trends in mortality for cancers of the colon/rectum, breast, lung/bronchus, and prostate are found to be responsible for 75% of the decreasing trend in cancer mortality. CONCLUSIONS: It is possible to partition an overall trend in age-adjusted rates under the assumption that it and the trends for all mutually exclusive and exhaustive subgroups of interest are linear. PMID- 10680728 TI - Kidney cancer and occupational exposure to asbestos: a meta-analysis of occupational cohort studies. AB - OBJECTIVE: To study the risk of kidney cancer following asbestos exposure. METHODS: We carried out a meta-analysis of the results of cohort studies of workers predominantly exposed to asbestos. We contacted the authors of 70 cohort studies; published results were available from the reports of 10 cohorts; we obtained the relevant information for an additional 27 cohorts. RESULTS: The studies included in the analysis comprised a total of 169 kidney cancer deaths and 69 incident cases. The overall pooled standardized mortality ratio (SMR) of kidney cancer was 1.1, with a 95% confidence interval (95% CI) of 0.9-1.3. The pooled SMR was higher for workers with undefined asbestos exposure (SMR 1.2, 95% CI 0.9-1.6) than for workers with either predominant chrysotile (SMR 0.9, 95% CI 0.7-1.3) or some amphibole (SMR 0.96, 95% CI 0.6-1.5) exposure. Studies with published results had higher SMRs than studies for which information was obtained from the authors. Studies with high asbestos exposure and an elevated SMR of lung cancer tended to show an increased risk of kidney cancer. CONCLUSIONS: It is unlikely that asbestos exposure is responsible for an important increase in kidney cancer risk; however, high asbestos exposure might entail a small increase in risk. PMID- 10680729 TI - A multicenter case-control study of diet and lung cancer among non-smokers. AB - OBJECTIVE: We have examined the role of dietary patterns and specific dietary nutrients in the etiology of lung cancer among non-smokers using a multicenter case-control study. METHODS: 506 non-smoking incident lung cancer cases were identified in the eight centers along with 1045 non-smoking controls. Dietary habits were assessed using a quantitative food-frequency questionnaire administered by personal interview. Based on this information, measures of total carotenoids, beta-carotene and retinol nutrient intake were estimated. RESULTS: Protective effects against lung cancer were observed for high consumption of tomatoes, (odds ratio (OR) = 0.5; 95% confidence interval (CI) 0.4-0.6), lettuce (OR = 0.6; 95% CI 0.3-1.2), carrots (OR = 0.8; 95% CI 0.5-1.1), margarine (OR = 0.7; 95% CI 0.5-0.8) and cheese (OR = 0.7; 95% CI 0.5-1.0). Only weak protective effects were observed for high consumption of all carotenoids (OR = 0.8; 95% CI 0.6-1.0), beta-carotene (OR = 0.8; 95% CI 0.6-1.1) and retinol (OR = 0.9; 95% CI 0.7-1.1). Protective effects for high levels of fruit consumption were restricted to squamous cell carcinoma (OR = 0.7; 95% CI 0.4-1.2) and small cell carcinoma (OR = 0.7; 95% CI 0.4-1.2), and were not apparent for adenocarcinoma (OR = 0.9; 95% CI 0.6-1.3). Similarly, any excess risk associated with meat, butter and egg consumption was restricted to squamous and small cell carcinomas, but was not detected for adenocarcinomas. CONCLUSIONS: This evidence suggests that the public health significance of increasing vegetable consumption among the bottom third of the population would include a reduction in the incidence of lung cancer among lifetime non-smokers by at least 25%, and possibly more. A similar protective effect for increased fruit consumption may be present for squamous cell and small cell lung carcinomas. PMID- 10680730 TI - Thyroid cancer in French Polynesia between 1985 and 1995: influence of atmospheric nuclear bomb tests performed at Mururoa and Fangataufa between 1966 and 1974. AB - BACKGROUND: Between 1966 and 1974, France performed 41 atmospheric nuclear weapon tests in the Mururoa and Fangataufa atolls in French Polynesia. METHODS: We performed a geographic analysis of thyroid cancer incidence, using data from the cancer registry of French Polynesia, medical evacuation files, insurance records and hospital and pathology laboratory files. RESULTS: A total of 153 thyroid cancers were diagnosed between 1985 and 1995 in the population born before 1976 and residing in French Polynesia. The incidence of thyroid cancer was 2-3 times larger in French Polynesia than in Maoris of New Zealand and Hawaiians of Hawaii. Based on few cases, a nonsignificant (p = 0.1) increase with decreasing distance between Mururoa and the birth place was observed in women born between 1950 and 1975 for thyroid cancer. CONCLUSION: Because the difference between Polynesian and reference populations was not larger for Polynesians who were children during the tests than for Polynesians born earlier; as would be expected in the case of radioiodine contamination, the high thyroid cancer rates in French Polynesia could hardly be attributed to radioiodine fallout. Nevertheless, a surveillance of the population born close to Mururoa is necessary to confirm or deny the existence of a higher risk of thyroid cancer in this population. PMID- 10680731 TI - Premorbid diet in relation to survival from prostate cancer (Canada). AB - OBJECTIVES: To examine the associations between prediagnostic energy, fat, and vitamin A intake and survival from prostate cancer. METHODS: Two hundred and seven cases of prostate cancer from Toronto and 201 cases from Vancouver provided diet histories at diagnosis between 1989 and 1992 and were followed for survival from prostate cancer. After exclusions for various reasons, 263 cases (135 from Toronto, 128 from Vancouver) were analyzed in Cox proportional hazards models. RESULTS: Following adjustments for clinical stage, histologic grade, and other factors, significantly lower risks of dying from prostate cancer in the highest compared with the lowest tertiles of monounsaturated fat intakes were observed in each city and in the combined city analyses (combined cities: hazard ratio [HR] = 0.3; 95% confidence interval (CI) = 0.1-0.7). Survival from prostate cancer was significantly better for cases in the highest tertile of energy intake in Toronto (HR = 0.1; CI = 0.01-0.6) in contrast to that in Vancouver where these cases did relatively worse (HR = 2.6; CI = 0.6-10.7). Other nutrients were either not consistently or not significantly associated with prostate cancer survival in the two cities. CONCLUSIONS: This bi-center cohort study observed a consistent and significant inverse association between the premorbid intake of monounsaturated fat and risk of death from prostate cancer. The inconsistent results for energy intake between cities could potentially be attributed to non-respondent bias in Toronto. PMID- 10680732 TI - Prostate cancer in Saskatchewan Canada, before and during the PSA era. AB - OBJECTIVES: The purpose of this study was to describe changes in the incidence of prostate cancer and in survival of diagnosed cases, as well as prostate cancer specific mortality, during a period spanning the introduction of prostate specific antigen (PSA) testing in Saskatchewan in 1990. METHODS: All cases of neoplasms of the prostate (ICD-O = C61.9) diagnosed in Saskatchewan from 1970 to 1997 inclusive, were identified in the Saskatchewan Cancer Registry. A subgroup of adenocarcinomas was defined for further study. Age-adjusted and age-specific incidence rates, and actuarial and relative survival were calculated according to time period of diagnosis. Age-adjusted and age-specific mortality rates from prostate cancer were also calculated for each time period, using Vital Statistics data. RESULTS: The age-adjusted incidence of prostate cancer was 60.5 per 10(5) in 1970, rising gradually to 101.5 per 10(5) in 1989. In 1990, incidence rose much more sharply, reaching a peak of 163.1 per 10(5) in 1993, after which it began to fall. This sharp increase coincided with the introduction and increasing use of the PSA test in the province. Relative survival of prostate cancer patients was stable from the late 1970s through the 1980s, then improved markedly in the 1990-94 period. After the introduction of the PSA test, the relative risk of death for prostate cancer patients was only about 60% of what it had been throughout the previous 15 years. Prostate cancer-specific death rates did not change from the early 1980s to the end of the study period. CONCLUSIONS: The above data are consistent with earlier diagnosis of prostate cancer due to PSA screening. Because mortality has not yet changed, it is premature to recommend widespread screening of asymptomatic men. PMID- 10680733 TI - Research questions concerning insulin-like growth factor-1. AB - The Harvard Center for Cancer Prevention (HCCP) convened a workshop on June 4, 1999 in Boston, MA. The objectives of the meeting were to briefly review the current state of knowledge in the area of insulin-like growth factor-1 physiology as related to cancer risk, to define the area's major remaining gaps in knowledge, and lastly to discuss research opportunities. PMID- 10680734 TI - Illnesses caused by smoking cigarettes. PMID- 10680735 TI - Response to the letter by Maria Feychting and Anders Ahlbom (Cancer Causes and Control 10: 637, 1999.) Why is the cancer pattern so different among visually impaired persons in Finland and Sweden? PMID- 10680736 TI - Newcastle disease virus (NDV): brief history of its oncolytic strains. AB - BACKGROUND: While genetically engineered viruses are now being tested for the virus therapy of human cancers, some naturally occurring viruses display unmatched oncolytic activity. Newcastle disease virus (NDV) excels as an oncolytic agent. OBJECTIVES: As its virulence versus attenuation can be explained on molecular biological bases, it may be possible to develop or select highly oncolytic strains of NDV without adverse toxicity. STUDY DESIGN: Questions are posed as to the mechanisms of viral oncolysis, the appropriateness of tests to predict oncolytic activity of a given NDV strain and the best modes of administration for oncolytic effects. Answers are provided based on specific data or on considerations drawn from experience (the authors use NDV oncolysates to immunize against melanoma and kidney carcinoma) or from analogous clinical situations (therapeutic use of mumps or measles viruses). RESULTS AND CONCLUSIONS: NDV oncolysates probably suit better for immunotherapy (providing also active tumor-specific immunization) than massive repeated inoculations of NDV strains, especially when the NDV strain used is not proven to be oncolytic by appropriate pre-clinical tests. PMID- 10680737 TI - The possible role of human cytomegalovirus (HCMV) in the origin of atherosclerosis. AB - BACKGROUND: The biological properties of some herpesviruses such as the ability of latent persistency in the host cells and the presence of viral DNA in atherosclerotic lesions, suggest the possible role of herpesviruses in the development of atherosclerosis. Although many authors proved the presence of viral DNA in arterial wall tissue, the role of herpesviruses in the origin and progress of atherogenesis still remains unclear. OBJECTIVES: The aim of this study was to confirm the presence of viral DNA in arterial wall and to associate the presence of these viruses with the development of atherosclerosis in patients with ischemic heart disease (IHD). STUDY DESIGN: A possible role of HCMV, EBV and HHV6 in the development of atherosclerosis was tested in 244 IHD patients and 87 coronarographically negative controls. The presence of viral DNA in aortic and venous walls, as well as in a peripheral blood samples was tested by the use of polymerase chain reaction (PCR) accompanied by, immunological tests for anti virus antibodies IgM and IgG types for all experimental groups. RESULTS: The genomic DNA of HCMV was found in 76 and 59%, DNA of EBV in 59 and 50%, and DNA of HHV6 in 0.08 and 0.0%, of arterial walls of IHD patients and non-ischemic control group, respectively. No viral DNA was found in venous samples. Significant association (P < 0.01) has been proved between CMV infection and IHD. CONCLUSIONS: Our results suggest that HCMV and EBV can be found in the arterial wall, so that the arterial wall could be a potential site of persistency of those viruses. We also proved a significant association between the presence of HCMV DNA in aortic walls and atherosclerosis. Despite of the high genetic and biological similarity between CMV and HHV6 no substantial role of HHV6 in atherosclerosis has been proved. PMID- 10680738 TI - Recent advances in the therapy and prevention of CMV infections. AB - The introduction of highly active antiretroviral therapy (HAART) for HIV has had a major impact on the treatment of CMV disease in HIV-infected individuals. There is mounting evidence that in patients with CMV retinitis who have a sustained response to HAART, CMV maintenance treatment can be discontinued without relapse of retinitis. In HAART-naive individuals with newly diagnosed CMV retinitis, the optimal timing for the initiation of HAART relative to the start of anti-CMV treatment is currently unknown. New local therapies for CMV retinitis (e.g. ganciclovir implant, the new antisense compound fomivirsen) provide treatment options in situations where high local drug delivery is warranted. A treatment algorithm for CMV disease in the HAART era is proposed. In the transplant setting, ganciclovir and foscarnet remain the major compounds used for treatment of CMV disease. In marrow and stem cell transplant recipients, CMV pneumonia still carries a high mortality. Ganciclovir in combination with CMV-specific immunoglobulin or regular intravenous IG remains the treatment of choice for CMV pneumonia; extended antiviral maintenance for several months is recommended in patients with continued immunosuppression. Preemptive treatment based on virologic markers (e.g. pp65 antigenemia, CMV DNA) has been very successful in reducing the incidence of early CMV disease in the transplant setting. The duration of preemptive treatment should be guided by the underlying immunosuppression and virologic markers. Late CMV disease is a challenge in marrow and stem cell transplant recipients, and occurs increasingly in highly immunosuppressed solid organ transplant recipients as well. Recent advances in prophylaxis strategies include oral ganciclovir for liver transplant recipients and valacyclovir for kidney transplant recipients. PMID- 10680739 TI - Rapid detection of respiratory viruses by centrifugation enhanced cultures from children with acute lower respiratory tract infections. AB - BACKGROUND: Acute respiratory tract infection (ARI) is the major cause of morbidity and mortality in young children in developing countries. Information on viral aetiology in ARI in India is very limited. OBJECTIVE: The aim of the study was to define the role of viruses in acute lower respiratory tract infections (ALRTI) in children in India using centrifugation enhanced cultures followed by indirect immunofluorescence (IIF). STUDY DESIGN: Nasopharyngeal aspirates (NPAs) were collected from children from September 1995 to April 1997, attending paediatric clinic of All India Institute of Medical Sciences (AIIMS) with symptoms of ALRTI. Virus isolation was done by centrifugation enhanced cultures using HEp-2, LLC-MK2 and MDCK cells. The viruses were identified at 24-48 h post inoculation by IIF staining using monoclonal antibodies to respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus and adenovirus. RESULTS: Of 200 NPA samples, 89 (44.5%) were positive for one or more viral pathogens. RSV was detected in 34 (17%) of all ALRTI cases followed by influenza viruses in 29 (14.5%), PIVs in 23 (11.5%) and adenoviruses in three (1.5%). In 79 children with bronchiolitis, RSV was most frequently isolated (25%) pathogen, while in bronchopneumonia cases (101) the most common viral pathogen was influenza virus (17%). In eight cases (4%) of ALRTI dual infections were detected. In 100 NPA specimens IIF staining on direct cell smears was carried out and viruses were detected in only 17%. RSV and influenza virus infection peaked from September to December, where as PIV infections were more frequent from January to April. CONCLUSION: Respiratory viruses accounted for 44.5% of cases of ALRTI in India and the results of viral aetiology could be given in 24-48 h using centrifugation enhanced cultures. RSV was the most common viral agent associated with ALRTI in children under 5 years of age with greater association with bronchiolitis. PMID- 10680740 TI - Seroprevalence of viral childhood infections in Eritrea. AB - BACKGROUND: The seroprevalence of viral childhood infections in Africa has not been thoroughly investigated. The relatively recently discovered human parvovirus B19 (B19) and human herpesvirus 6 (HHV-6) have received particularly little attention. OBJECTIVE: To investigate the seroprevalence of viral childhood infections in different Eritrean populations and to define groups at high risk for infection. STUDY DESIGN: Five population groups in Eritrea have been examined to define the prevalence of specific antibodies to several childhood viruses. The study population of more than 400 persons consisted of children, pregnant women, female sex workers and members of a secluded tribe called Rashaida. RESULTS: All groups showed a high prevalence of antibodies to measles and HHV-6 (> 85%). For rubella, the seroprevalence was very high in all adult groups (93-99%) except the Rashaida group (71%). The mumps prevalence was surprisingly low in the Rashaida group (29%) compared to 46-85% in the other adults. Late encounter of mumps and rubella was also observed among the Rashaidas. The pattern of antibodies to B19 showed a higher seroprevalence in all groups (56-91%) compared to what has been reported from the western world. CONCLUSION: The findings represent what might be expected in an unvaccinated population. The exception was the Rashaidas, which had low seroprevalences and late encounter of mumps and rubella. This is of importance because it makes this tribe vulnerable to these infections, which are associated with complications when acquired in adult age. Also noteworthy is the high frequency of antibodies to HHV-6 and particularly B19 in all groups, indicative of an early encounter of both these viruses. PMID- 10680741 TI - No apparent effect of cidofovir in epidermodysplasia verruciformis. AB - This paper reports the failure of a patient suffering from Epidermodysplasia verruciformis, characterised by widespread infection of the skin with human papillomaviruses, to respond to topical and systemic treatment with the antiviral agent, Cidofovir, despite its previously demonstrated effectiveness against a range of different papillomavirus-associated conditions. PMID- 10680742 TI - Comparison of six nucleic acid extraction methods for detection of viral DNA or RNA sequences in four different non-serum specimen types. AB - BACKGROUND: Extraction of viral nucleic acids from serum samples is widely used in diagnostic pathology tests. However, the heterogeneous nature of non-serum samples may contribute to variations in the yields of viral nucleic acids with different extraction methods and specimen types. OBJECTIVES: Six different methods were compared for optimal extraction of viral DNA or RNA from four types of non-serum specimens. STUDY DESIGN: The DNA viruses used were herpes simplex virus and cytomegalovirus. The RNA viruses were poliovirus, rotavirus and small round structured virus. The specimens used were from respiratory, genital, faecal and peripheral blood mononuclear cell samples. The extracted nucleic acids were amplified by PCR and detected in an enzyme immunoassay using digoxygenin-labelled amplicons. RESULTS AND CONCLUSIONS: For extraction of viral DNA, the phenol chloroform method yielded the highest amount of DNA as judged by endpoint titration. The three methods compared for extraction of viral RNA used guanidine isothiocyanate and the QiaRNA kit was shown to yield the highest amount of viral RNA. PMID- 10680743 TI - Asymptomatic shedding of herpes simplex virus into the oral cavity of patients with atopic dermatitis. AB - BACKGROUND: Asymptomatic shedding of herpes simplex virus (HSV) into the oral cavity is one of the important sources of the infection. OBJECTIVES: We anticipated that the oral cavity is one of the possible sites of initial recurrence in patients with recurrent-type eczema herpeticum. MATERIALS AND METHODS: Oral swabs were collected from 24 patients with atopic dermatitis aged from 13 to 29 years, and as a control population, from seven patients with skin diseases other than atopic dermatitis aged from 16 to 29 years. We tried to detect HSV DNA by the polymerase chain reaction method. RESULTS: HSV DNA was detected in eight out of 31 specimens from 15 patients with facial lesions of atopic dermatitis, and in one of 15 specimens from nine patients without facial atopic dermatitis, and in two of ten specimens from seven patients with other skin diseases. However, these differences are not statistically significant. Moreover, no patients developed eczema herpeticum during the period of this study. CONCLUSIONS: These results suggest that facial atopic dermatitis may not lead to oral HSV shedding, and there may be little opportunity for the virus in the oral cavity to contaminate other skin sites in adolescents and adults. PMID- 10680744 TI - Hepatitis E virus in Cuba. AB - The hepatitis E virus (HEV) has a global distribution and is known to have caused large waterborne epidemics of icteric hepatitis. The transmission is primarily fecal-oral. Some reports have suggested parenteral transmission of HEV from its association to hepatitis B or hepatitis C infection, or due to the development of hepatitis E after blood transfusion. Though most of the developing countries in Asia and Africa have been shown to be endemic for HEV infection, studies in the Latin American countries have been limited to Mexico, Brazil and Venezuela. We have developed an enzyme immunoassay (EIA) for IgM and IgG antibodies to a recombinant protein containing antigenic epitopes of the ORF3 region of the HEV. This system, as well as a commercial kit that includes ORF2 and ORF3 antigenic epitopes, were used to study the prevalence of anti-HEV antibodies in a sample of Cuban blood donors, acute hepatitis cases and individuals subjected to plasmapheresis. The incidence of anti-HEV IgM was compared with other viral hepatitis markers. Our findings suggest that infections due to HEV are an important viral cause of sporadic hepatitis in Cuba, and that HEV is endemic to this region of the world. PMID- 10680745 TI - Concanavalin A-induced hepatitis in mice is prevented by interleukin (IL)-10 and exacerbated by endogenous IL-10 deficiency. AB - One single intra-venous (i.v.) injection of Concanavalin A (Con A) into mice provokes a cell-mediated immunoinflammatory hepatitis. We have presently evaluated the immunopharmacological effects of exogenous interleukin (IL)-10 and the role of endogenous IL-10 in this model by using exogenous IL-10, anti-IL-10 monoclonal antibody (mAb) and mice with disrupted IL-10 gene (IL-10 KO mice). Whilst exogenous IL-10 administered in a prophylactic (1 h prior to Con A) and even "early" therapeutic fashion (30 min after Con A) reduced the elevation of transaminase activities in plasma in a dose-dependent manner, observed in control mice, these biochemical markers of liver injury were significantly increased both in IL-10 KO mice as well as in those receiving anti-IL-10 mAb. Interestingly, doses of Con A lower than 20 mg/kg that were only capable of inducing slight serological signs of hepatitis in mice, exerted marked hepatitic effects when administered to either anti-IL-10 mAb-treated mice or to IL-10 KO mice. The disease modulating effects of exogenous IL-10 and either genetical or pharmacologically-induced IL-10 deficiency were associated with profound and opposite modifications of the Con A-induced increase in the circulating levels of IFN-gamma and TNF-alpha. Relative to control animals, the blood levels of these cytokines were diminished in IL-10-treated mice and augmented in both IL-10 KO mice and anti-IL-10 mAb-treated mice. These results prove the physiological antiinflammatory role of endogenous IL-10 in Con A induced hepatitis and the beneficial effects of IL-10 treatment to prevent this condition. PMID- 10680746 TI - Serum soluble HLA-DR antigens in autoimmune hepatitis. AB - To investigate the significance of HLA-class II, especially DR antigens, in autoimmune hepatitis (AIH), the serum concentrations of soluble HLA-DR antigen (sDR) were measured in 16 patients with AIH. The expression of HLA-DR antigens in the liver tissues of AIH patients was also studied by immunohistochemistry. AIH at diagnosis showed markedly higher serum sDR levels than controls, in which the liver tissues exhibited positive staining of HLA-DR antigens. Seven patients received corticosteroid therapy, in whom the serum sHLA-DR concentration was reduced dramatically from activated to remission stage. In sequentially follow-up cases, sDR correlated well with the disease activity, and also with the change of surface DR expression in the liver. A single major band with a molecular size of 60 kDa was detected, both in patient's sera and in normal control sera, by Western blotting. In conclusions, serum sHLA-DR level could be a marker reflecting immunological activity of the disease. PMID- 10680747 TI - The 5S rRNA is associated with Ro60 ribonucleoprotein and is co-precipitated with hYRNAs by anti-Ro antibodies. AB - Ro particles are conserved molecules that contain a YRNA and various Ro proteins, which are recognized by autoimmune sera from patients with lupus erythematosus or Sjogren's syndrome. The Ro60 ribonucleoprotein (RNP) forms complexes with certain 5S rRNAs, in such a manner that Ro60 could participate in the control of 5S rRNA production. The present studies were carried out to explore the interaction of Ro components, and to address the question whether Ro60 RNP binds simultaneously 5S rRNA and hYRNA. Anti-Ro60 antibodies were used to immunoprecipitate the RNA. Immunoprecipitates were reverse transcribed with specific oligonucleotides and the resulting cDNAs from 5S and hY4 were amplified by PCR. We found that 5S rRNA is complexed with hY4 and hY5 RNAs by means of the Ro60 RNP. Moreover, by in situ hybridization assays we were able to demonstrate that these molecules have a similar nuclear distribution. According to these results, it seems reasonable to assume that the Ro60 protein could be involved in ribosome assembly. PMID- 10680748 TI - Serial changes in the galactosylation of autoantibodies and serum IgG in autoimmune haemolytic anaemia. AB - A number of systemic autoimmune diseases are associated with increased levels of the agalactosyl (G0) IgG isoforms that lack a terminal galactose from the C(H)2 domain oligosaccharide. The aims were to determine whether there are also persistently high levels of G0 autoantibodies or serum IgG in autoimmune haemolytic anaemia (AIHA), and whether any changes in galactosylation over time are related to the course of disease. Autoantibodies eluted from red blood cells, and serum IgG, were obtained from a patient with chronic AIHA over a 21 month period, and the degree of galactosylation measured using a lectin-binding assay. There were wide fluctuations in the galactosylation of autoantibody and serum IgG, but these changes were unrelated to the severity of the anaemia. The galactosylation of autoantibody and serum IgG varied independently, and the autoantibodies were preferentially G0 in comparison with serum IgG in only half of the serial samples. We conclude that AIHA differs from other, systemic autoimmune conditions in that high levels of G0 autoantibodies or serum IgG are not persistent, and that changes in galactosylation do not parallel the course of disease. PMID- 10680749 TI - Anti-Beta 2-glycoprotein I antibody isotype and IgG subclass in antiphospholipid syndrome patients. AB - Beta 2-glycoprotein I (beta2-GPI) is an antigenic target recognised by antiphospholipid antibodies found in association with the antiphospholipid syndrome (APS). In this study, the prevalence of Immunoglobulin M (IgM) and IgA anti-beta2-GPI antibodies was examined in APS patients and compared with IgG antibodies. In addition the value of measuring antibody isotypes and IgG subclass was investigated in the laboratory diagnosis of APS. A solid phase enzyme linked immunosorbent assay was established to measure IgG, IgM and IgA and IgG subclass antibodies to beta2-GPI in patients with APS and a variety of other thrombotic and non-thrombotic disorders. Raised levels of IgM anti-beta2-GPI antibodies were observed in 65% of patients with APS, 21% with systemic lupus erythematosus (SLE), 23% with rheumatoid factor, 4% with stroke, 5% carotid artery stenosis (CAS), 17% with a biological false positive serology for syphilis, 43% with infectious mononucleosis (IM) and 27% with human immunodeficiency virus (HIV). The median value for IgM antibodies to beta2-GPI for all these groups ranged from 2 to 7 arbitrary units (AU). Elevated levels of IgA antibodies to beta2-GPI were found in patients with APS (47%), SLE (13%), rheumatoid factor (26%), CAS (48%), stroke (25%), VDRL false positive serology for syphilis (33%), IM (47%) and HIV (7%). The median value of IgA antibodies to beta2-GPI in all of these groups ranged from 2 to 4 AU. Conversely the median value for IgG anti-beta2-GPI in APS patients was 112 AU compared to 1-4 AU in the other conditions examined. The presence of IgM and IgA antibodies to beta2-GPI was much less specific and sensitive for APS than IgG, with raised levels of these isotypes seen in a variety of thrombotic and non-thrombotic disorders. Elevated levels of IgG1, IgG2, IgG3 and IgG4 antibodies to beta2-GPI were detected in APS patients. While all four IgG anti-beta2-GPI antibody subclasses were represented in APS patients there appeared to be a significant overall skewing towards to the IgG2 subclass. PMID- 10680750 TI - Reduced in vitro production of interferon-gamma, interleukin-4 and interleukin-12 and increased production of interleukin-6, interleukin-10 and tumour necrosis factor-alpha in systemic lupus erythematosus. Weak correlations of cytokine production with disease activity. AB - Production of cytokines in unstimulated and mitogen-stimulated cultures were evaluated by ELISPOT in 34 SLE patients with low to moderate disease activity and 23 healthy controls. Significantly reduced production of IFN gamma, IL4 and IL12 and significantly increased production of IL6, IL10 and TNF alpha were found in patients with SLE. Regression analysis revealed that production of all six cytokines tended to decrease with increasing disease activity, but negative correlation with SLEDAI was significant (p < 0.05) only for PHA-stimulated IL4, unstimulated and PHA-stimulated IL10 and SAC-stimulated IL6. Negative correlation of stimulated and unstimulated IL6 and TNF alpha production with anti-DNA antibody levels were also significant. PMID- 10680751 TI - Expression of HSP-65 in jejunal epithelial cells in patients clinically suspected of coeliac disease. AB - BACKGROUND: Coeliac disease (CD) can be classified both clinically and biologically an autoimmune disease. A close relationship obtains between heat shock proteins (HSPs) and numerous autoimmune diseases. HSPs are overexpressed when protecting the host against environmental insult. We sought here to establish whether dietary gluten is such a stress stimulus in patients clinically suspected of CD, and whether the expression of HSP-65 associates with densities of intraepithelial gammadelta+ T cells and/or with expression of mucosal HLA-DR. METHODS: Seventy-eight children with clinical suspicion of CD underwent a jejunal biopsy. Monoclonal antibodies were used to stain jejunal epithelial HSP-65, intraepithelial lymphocytes and mucosal HLA-DR. Serum IgA-class endomysial autoantibodies (EMA) were measured by an indirect immunofluorescence method. CD susceptibility HLA DQA1*0501 and DQB1*0201 alleles (HLA DQ2) were determined. RESULTS: Enhanced expression of epithelial cell mitochondrial HSP-65 was found in 80% (16/20) of coeliacs and in 24% (14/58) of children excluded for the disease, but in only 7% (2/28) of control subjects (p < 0.001, p = 0.049, respectively). Children with enhanced expression of HSP-65 had significantly higher gammadelta+ T cell densities than those with normal HSP-65 expression. A clear association between HSP-65 and serum IgA-class EMA were also ascertained in patients with normal jejunal mucosal morphology. HLA DQ2 positivity did not correlate with the HSP-65 expression. CONCLUSIONS: Gluten might be an environmental insult not only in CD patients but also in some patients excluded for the disease on biopsy. Enhanced expression of epithelial cell stress proteins might be an indicator of such an insult. PMID- 10680752 TI - Factors involved in the pathogenesis of neutrophilic vasculitis in MRL/Mp-lpr/lpr mice: a model for human microscopic angiitis. AB - Anti-neutrophil cytoplasm antibodies (ANCA) directed against myeloperoxidase (MPO) are detected in patients with microscopic angiitis. Human MPO autoantibodies stimulate neutrophil degranulation in vitro and are thought to be pathogenic. We have previously shown that MRL-lpr mice with MPO autoantibodies have a higher incidence of vasculitis than their seronegative littermates. The aim of the present study is to determine the relationship between MPO autoantibodies and microscopic angiitis. The neutrophil binding properties of anti-MPO monoclonal antibodies (mAbs) from MRL-lpr mice were tested using murine heterophils (neutrophils) present in blood and induced peritoneal exudates. MRL anti-MPO mAbs selectively bind activated neutrophils which express MPO in vitro. The pathogenicity of an IgG2b anti-MPO mAb, C6, was investigated in vivo. Anti MPO mAb, C6 was administered to young MRL mice which had been primed with exogenous TNF alpha to induce neutrophil activation and expression of MPO. Neutrophilic vasculitis similar to microscopic angiitis occurred in 33% of MRL mice which had been treated with anti-MPO mAb. The lesions were mainly restricted to sites of previous endothelial insult which suggests an active role for injured endothelium in this pathology. PMID- 10680753 TI - Amygdala, hippocampus and discriminative fear conditioning to context. AB - Various measures of fear have been shown to condition to a fearful context with different acquisition rates (Antoniadis EA, McDonald RJ. Fear conditioning to context expressed by multiple measures of fear in the rat, Behav Brain Res 1999;101(1):1-14). Freezing, locomotion, urination and preference are 'fast' measures of fear in that they discriminatively condition to context after a single training session, while ultrasonic vocalizations and defecation are 'slow' measures of fear given that they condition following three training sessions. In the present experiment we sought to assess the contribution of the amygdala and the hippocampus in this form of learning. Existing views differ on the degree of involvement of each memory structure. This discord probably emerges from the common use of non-discriminative paradigms and the assessment of a single measure of fear. With the use of a discriminative paradigm and the assessment of multiple measures of fear, results indicate that the amygdala is a memory structure that selectively mediates the conditioning of heart rate, and the hippocampus selectively mediates the conditioning of defecation and body temperature. The conditioning of preference, locomotion, freezing and ultrasonic vocalizations, necessitate the participation of both memory structures while the conditioning of urination does not seem to require the participation of either the hippocampus or the amygdala. The proposed view ascribes an equal role in fear conditioning to both the amygdala and the hippocampus. PMID- 10680754 TI - Visually guided locomotion and computation of time-to-collision in the mongolian gerbil (Meriones unguiculatus): the effects of frontal and visual cortical lesions. AB - Past research has indicated that many species use the time-to-collision variable but little is known about its neural underpinnings in rodents. In a set of three experiments we set out to replicate and extend the findings of Sun et al. (Sun H J, Carey DP, Goodale MA. Exp Brain Res 1992;91:171-175) in a visually guided task in Mongolian gerbils, and then investigated the effects of lesions to different cortical areas. We trained Mongolian gerbils to run in the dark toward a target on a computer screen. In some trials the target changed in size as the animal ran toward it in such a way as to produce 'virtual targets' if the animals were using time-to-collision or contact information. In experiment 1 we confirmed that gerbils use time-to-contact information to modulate their speed of running toward a target. In experiment 2 we established that visual cortex lesions attenuate the ability of lesioned animals to use information from the visual target to guide their run, while frontal cortex lesioned animals are not as severely affected. In experiment 3 we found that small radio-frequency lesions, of either area VI or of the lateral extrastriate regions of the visual cortex also affected the use of information from the target to modulate locomotion. PMID- 10680755 TI - The Dalila effect: C57BL6 mice barber whiskers by plucking. AB - Group-housed laboratory mice are frequently found with their whiskers and facial hair removed. It has been proposed that dominant mice are responsible for barbering the hair of the recipient (the Dalila effect), and early studies suggest that the hair is removed by nibbling. In the present study, pairs of C57BL6 mice, composed of a barber and recipient, were separated to allow hair to regrow. The animals were then placed together in an observation box and their social behavior was videorecorded. The videorecording was subjected to frame-by frame analysis. Barbering was found to occur during acts of mutual grooming. During grooming, one member of a mouse pair removed the vibrissae of the conspecific and did so by grasping individual whiskers with the incisors and plucking them out. Although plucking appeared 'painful', recipients were passive in accepting barbering, and even pursued conspecifics for further grooming. Other measures indicated that barbers were heavier than recipients and brain weights were not different. Although cortical barrel fields appeared normal to cytochrome oxidization and zinc staining, Golgi analysis of layer three, barrel-field basilar dendrites indicated changes in cell morphology. The results are discussed in relation to the hypothesis that barbering is an expression of social dominance, the origins of the barbering behavior, and the consequences of barbering on brain function. PMID- 10680756 TI - Use of a delayed non-matching to position task to model age-dependent cognitive decline in the dog. AB - Spatial learning and memory in young and old dogs was studied in a series of experiments using a delayed non-matching to position (DNMP) paradigm. Past research from our laboratory has suggested that aged dogs perform more poorly on a version of the DNMP task compared to young dogs [Head et al., Spatial learning and memory as a function of age in the dog, Behav. Neurosci. 1995;109(5):851 585]. We have now extended these findings by testing a large number of dogs on three different variations of the DNMP paradigm to evaluate different aspects of spatial learning and memory. Our results indicate that: (1) aged dogs show impaired spatial learning compared to young dogs, (2) aged dogs display spatial working memory deficits compared to young dogs, (3) young dogs have a greater maximum working spatial memory capacity than old dogs and (4) we can use the DNMP paradigm to cognitively categorize different subsets of aged dogs. These data indicate that the DNMP paradigm can serve as a valuable tool to evaluate age dependent cognitive dysfunction in the canine. PMID- 10680757 TI - Effects of NMDA receptor blockers on cocaine-stimulated locomotor activity in mice. AB - Effects of MK-801 and ketamine, N-methyl-D-aspartate (NMDA) receptor blockers, on cocaine-stimulated locomotor activity were investigated in male Swiss-Webster mice. MK-801 (0.25, 0.5, 1.0 and 2.5 mg/kg), ketamine (10, 25 and 50 mg/kg) or saline was injected 20 min before cocaine (5, 10 and 20 mg/kg i.p.). Locomotor activity was measured for 30 min immediately following cocaine treatment. All doses of the drugs were also tested for ability to depress or stimulate locomotor activity in the naive (no cocaine-treated) mice. Cocaine produced a dose dependent increase in locomotor activity that was blocked dose-dependently by MK 801 or ketamine. The blockade by MK-801 was more prominent than by ketamine. Our results may suggest that cocaine-induced locomotor stimulation in mice is modulated via NMDA receptor mediated mechanisms. PMID- 10680758 TI - Transient focal cerebral ischemia induces sensorimotor deficits in mice. AB - Rodents have been extensively used for experimental stroke research with rat and gerbil the preferred species. With the advent of transgenesis and gene targeting the number of mutant mouse strains is rapidly increasing. Thus, mouse models of stroke will be of great importance in the analysis of genetic factors affecting stroke. Demonstrating long-term functional recovery is of paramount importance for the pharmacological evaluation of putative stroke therapies. In the present paper we induce mild focal cerebral ischemia by tandem occlusion of the right middle cerebral artery (MCA), via craniotomy, together with the common carotid artery for 45 min in C57BL/6 strain of mice. The effects of ischemia were evaluated acutely by MRI and long-term (> 3 weeks) sensorimotor functional deficits were analyzed using a number of behavioral paradigms including the rotorod, wire hang, horizontal surface approach, eye-closure reflex, and T-maze tests. Although the induced brain damage is mild we show that it leads to clearly detectable and significant sensorimotor defects associated with fine motor coordination, balance, and postural and sensory reflexes. We conclude that the applied behavioral tests will be useful in the analysis of stroke in mutant mice. PMID- 10680759 TI - D1 and D2 dopamine receptor agonists improve deficits in motor programming of cats with a 6-hydroxydopamine lesion in the A8 cell group. AB - Recently, it has been shown that a small 6-hydroxydopamine lesion in the A8 cell group of cats trained to walk on a treadmill produces long-lasting deficits (Arts and Cools, 1998, Behav. Neurosci. 112; pp. 102-105). Some deficits could be attributed to a hypofunction of A9 cells, that is a reduced ability to switch arbitrarily motor patterns, and other deficits to a hyperfunction of A10 cells, that is an improved ability to switch motor patterns with the help of cues. This experiment was repeated in this study and the elicited behavioural symptoms were systemically treated with the dopamine D1 receptor agonist SKF 81297 and dopamine D2 receptor agonist LY 171555. The results show that a cocktail of these agonists restored both the lesion-induced reduced ability to switch arbitrarily motor patterns and the lesion-induced increased ability to switch motor patterns with the help of cues, suggesting that this treatment restored the functional misbalance between the A9 and A10 cells. PMID- 10680760 TI - The mechanisms of movement preparation: a precuing study. AB - It is well known that precues about the possible locations of upcoming targets reduce the manual reaction time. The present study investigates at which stage of the sensorimotor system such precues act. Several light dots were displayed as a precue; one of them became the target, and subjects had to produce a manual response to the target as fast as possible. Exp. A varied independently the number of precues and the area of space they occupy; we found that the reaction time of pointing movements depended on spatial extent, but not on the number of choices. The outcome was similar in Exp. B, where subjects produced stereotyped 'tapping' movements irrespective of target position. Taken together, both findings support the view that precues act mainly at a stage concerned with the internal representation of space, rather than with response selection or movement preparation. The effects of precues were preserved when subjects fixated throughout the precuing period (Exp. C), but not when precue and target positions were uncorrelated (Exp. D). These findings do not support the alternative interpretations, that precues act by guiding the eyes into the vicinity of targets, or by elevating the subjects' arousal level. PMID- 10680761 TI - The effect of an external load on the force and timing components of mentally represented actions. AB - The chronometry of real and imagined movements was investigated in a group of eight subjects under varying conditions. The visually-guided pointing task was used to investigate the speed for accuracy trade-offs that occur as target size is varied for both real and imagined performance. The task was performed both with and without an external load of 2 kg. For the no-load condition and load conditions, the speed for accuracy trade-off for both real and imagined performance conformed to Fitts' law. Movement durations of real movements remained largely unaffected by the addition of the load, however, movement durations of imagined movements increased significantly with the addition of the load. These patterns of results suggest that the weight disrupted the force calculation component of imagined movements but not the relative timing. PMID- 10680762 TI - Sensory and motor aspects of the coital reflex in the spinal male rat. AB - In the present study the sensory and motor aspects of the coital reflex model in male rats were evaluated. Electromyographic reflex activity after mechanical stimulation of the urethra, penis and scrotal skin was recorded in all genital muscles. The possibility that a facilitatory mechanism of these muscular responses was located in the rat spinal cord was evaluated by removing the genital afferents. Results showed that urethral, penile and scrotal stimulation evoked the coital reflex in all genital muscles. Similarly, coital responses were obtained spontaneously in deafferentated animals. The parameters among the motor patterns evoked with the different mechanical stimuli were very similar. Parameters of spontaneous motor patterns were not importantly different from those obtained reflexively. A conspicuous difference between these responses was the presence of an after-discharge activity in genitally-stimulated animals. Additionally, it was found that this motor pattern can be exhausted with repeated stimulation. Spontaneous responses did not show the exhaustion phenomenon. Results are discussed in the context of the sensory-motor aspects of male sexual behaviour. PMID- 10680763 TI - Activation of Broca's area by syntactic processing under conditions of concurrent articulation. AB - Regional cerebral blood flow (rCBF) was measured with positron emission tomography (PET) when 11 subjects made plausibility judgments about written sentences that varied in their syntactic complexity. While making their judgments, subjects uttered the word "double" aloud at a rate of one utterance per second to inhibit their ability to rehearse the sentences. Blood flow increased in Broca's area when subjects made judgments about the more complex sentences. This result replicates and extends previous findings that blood flow increases in this region when subjects process complex syntax under no interference conditions. The results of this experiment provide strong evidence that the increase in blood flow seen in Broca's area in association with processing syntactically complex structures is not due to subvocal rehearsal of those structures, but rather results from processing syntactic forms themselves. PMID- 10680764 TI - Human brain areas involved in the analysis of auditory movement. AB - This work tests the hypothesis that a network of areas involving bilateral premotor cortex and right parietal cortex subserves the analysis of sound movement. The components of this network have been examined at the level of individual subjects in a study where 720 fMRI scans were acquired per subject. Additionally, the effect of movement direction was investigated by varying this property systematically. Linear sound ramps that are perceived as movement toward one side of the head or the other were used in an experiment in which the principal contrast was between movement, and a stationary control stimulus made up of identical component interaural phase and amplitude cues. In a group analysis, the network of bifrontal and right parietal areas suggested by previous work was confirmed. The frontal activation included both dorsal premotor activity in the region of the frontal eye fields and discrete ventral premotor activation in an area corresponding to primate areas for multimodal spatial analysis and motor planning. The right parietal activation included both superior and inferior parietal cortex. Analysis of the individual data showed a similar pattern of activation in each subject, with the greatest variability within the right parietal area. The pattern of activation did not vary when the direction of movement was varied, suggesting that both directions of movement are represented in the network we have demonstrated. PMID- 10680765 TI - Mathematical/computational challenges in creating deformable and probabilistic atlases of the human brain. AB - Striking variations in brain structure, especially in the gyral patterns of the human cortex, present fundamental challenges in human brain mapping. Probabilistic brain atlases, which encode information on structural and functional variability in large human populations, are powerful research tools with broad applications. Knowledge-based imaging algorithms can also leverage atlased information on anatomic variation. Applications include automated image labeling, pathology detection in individuals or groups, and investigating how regional anatomy is altered in disease, and with age, gender, handedness and other clinical or genetic factors. In this report, we illustrate some of the mathematical challenges involved in constructing population-based brain atlases. A disease-specific atlas is constructed to represent the human brain in Alzheimer's disease (AD). Specialized strategies are developed for population based averaging of anatomy. Sets of high-dimensional elastic mappings, based on the principles of continuum mechanics, reconfigure the anatomy of a large number of subjects in an anatomic image database. These mappings generate a local encoding of anatomic variability and are used to create a crisp anatomical image template with highly resolved structures in their mean spatial location. Specialized approaches are also developed to average cortical topography. Since cortical patterns are altered in a variety of diseases, gyral pattern matching is used to encode the magnitude and principal directions of local cortical variation. In the resulting cortical templates, subtle features emerge. Regional asymmetries appear that are not apparent in individual anatomies. Population based maps of cortical variation reveal a mosaic of variability patterns that segregate sharply according to functional specialization and cytoarchitectonic boundaries. PMID- 10680766 TI - Explicit and implicit neural mechanisms for processing of social information from facial expressions: a functional magnetic resonance imaging study. AB - The processing of changing nonverbal social signals such as facial expressions is poorly understood, and it is unknown if different pathways are activated during effortful (explicit), compared to implicit, processing of facial expressions. Thus we used fMRI to determine which brain areas subserve processing of high valence expressions and if distinct brain areas are activated when facial expressions are processed explicitly or implicitly. Nine healthy volunteers were scanned (1.5T GE Signa with ANMR, TE/TR 40/3,000 ms) during two similar experiments in which blocks of mixed happy and angry facial expressions ("on" condition) were alternated with blocks of neutral faces (control "off" condition). Experiment 1 examined explicit processing of expressions by requiring subjects to attend to, and judge, facial expression. Experiment 2 examined implicit processing of expressions by requiring subjects to attend to, and judge, facial gender, which was counterbalanced in both experimental conditions. Processing of facial expressions significantly increased regional blood oxygenation level-dependent (BOLD) activity in fusiform and middle temporal gyri, hippocampus, amygdalohippocampal junction, and pulvinar nucleus. Explicit processing evoked significantly more activity in temporal lobe cortex than implicit processing, whereas implicit processing evoked significantly greater activity in amygdala region. Mixed high-valence facial expressions are processed within temporal lobe visual cortex, thalamus, and amygdalohippocampal complex. Also, neural substrates for explicit and implicit processing of facial expressions are dissociable: explicit processing activates temporal lobe cortex, whereas implicit processing activates amygdala region. Our findings confirm a neuroanatomical dissociation between conscious and unconscious processing of emotional information. PMID- 10680767 TI - Frequency dependence of the functional MRI response after electrical median nerve stimulation. AB - Localizing sensorimotor areas with high resolution functional MRI is of considerable interest for a wide range of medical applications from the preoperative planning of neurosurgical interventions to determining the course of neuroplastic reorganisation after brain lesions. We examined the effect of the stimulation frequency on the blood oxygen level dependent (BOLD) fMRI response and on perfusion weighted fMRI using electrical median nerve stimulation at 5, 15, 40, and 100 Hz. BOLD fMRI was performed using a single shot gradient echo EPI sequence to acquire 15 contiguous slices. For the qualitative flow sensitive studies, a single slice inversion recovery prepared spin echo echoplanar sequence (IR-SE EPI) was used. In the primary sensorimotor cortex, a linear increase of the fMRI-BOLD response, affecting both the number of activated pixels and the amplitude of the signal changes, was seen with increasing stimulation frequencies. The qualitative in-flow sensitive studies, using the IR-SE EPI sequence, indicate that the tissue perfusion also increases over the same range of frequencies. This implicates that larger fMRI responses can be obtained if electrical median nerve stimulation is performed at higher frequencies. The results are compared with electrophysiological data, which show a decrease of the early somatosensory evoked potentials at higher frequencies. PMID- 10680768 TI - Search for compounds that inhibit the genotoxic and carcinogenic effects of heterocyclic aromatic amines. AB - Over the last 30 years approximately 160 reports have been published on dietary compounds that protect from the mutagenic and carcinogenic effects of heterocyclic aromatic amines (HAAs). In the first section of this review, the current state of knowledge is briefly summarized. Based on the evaluation of the available data, various protective mechanisms are described, and the use of different methodologies for the detection of protective effects is critically discussed. In most antimutagenicity studies (>70%) bacterial indicators (predominantly Salmonella strain TA98) were used, and about 600 individual compounds and complex mixtures have been identified that attenuate the effects of HAAs. The most frequently used in vivo method to detect protective effects are adduct measurements; anticarcinogenic dietary factors were identified by aberrant crypt foci assays and liver foci tests with rats. The mechanisms of protection include inactivation of HAAs and their metabolites by direct binding, inhibition of enzymes involved in the metabolic activation of the amines, induction of detoxifying enzymes, and interaction with DNA repair processes. The detection spectrum of conventional in vitro mutagenicity assays with metabolically incompetent indicator cells is limited. These procedures reflect only simple mechanisms such as direct binding of the HAAs to pyrroles and fibers. It has been shown that these compounds are also effective in rodents. More complex mechanisms, namely, interactions with metabolic activation reactions are not adequately represented in in vitro assays with exogenous enzyme homogenates, and false-negative as well as false-positive results may be obtained. More appropriate approaches for the detection of protective effects are recently developed test systems with metabolically competent cells such as the human Hep G2 line or primary hepatocytes. SCGE tests and DNA adduct measurements with laboratory rodents enable the detection of antigenotoxic effects in different organs, including those that are targets for tumor induction by the amines. Medium term assays based on aberrant crypt foci in colon and liver foci tests have been used to prove that certain compounds that prevented DNA damage by HAAs also reduced their carcinogenic effects. These experiments are costly and time consuming and, due to the weak induction capacity of the amines, only pronounced anticarcinogenic effects can be detected. Over the years, a large bulk of data on HAA protective compounds has accumulated, but only for a few (e.g., fibers, pyrroles, constituents of teas, and lactic acid bacteria) is there sufficient evidence to support the assumption that they are protective in humans as well. PMID- 10680769 TI - Health effects of endocrine-disrupting chemicals on wildlife, with special reference to the European situation. AB - Many wildlife species may be exposed to biologically active concentrations of endocrine-disrupting chemicals. There is strong evidence obtained from laboratory studies showing the potential of several environmental chemicals to cause endocrine disruption at environmentally realistic exposure levels. In wildlife populations, associations have been reported between reproductive and developmental effects and endocrine-disrupting chemicals. In the aquatic environment, effects have been observed in mammals, birds, reptiles, fish, and mollusks from Europe, North America, and other areas. The observed abnormalities vary from subtle changes to permanent alterations, including disturbed sex differentiation with feminized or masculinized sex organs, changed sexual behavior, and altered immune function. For most reported effects in wildlife, however, the evidence for a causal link with endocrine disruption is weak or nonexisting. Crucial in establishing causal evidence for chemical-induced wildlife effects appeared semifield or laboratory studies using the wildlife species of concern. Impaired reproduction and development causally linked to endocrine-disrupting chemicals are well documented in a number of species and have resulted in local or regional population changes. These include: Masculinization (imposex) in female marine snails by tributyltin, a biocide used in antifouling paints, is probably the clearest case of endocrine disruption caused by an environmental chemical. The dogwhelk is particularly sensitive, and imposex has resulted in decline or extinction of local populations worldwide, including coastal areas all over Europe and the open North Sea. DDE-induced egg shell thinning in birds has caused severe population declines in a number of raptor species in Europe and North America. Endocrine-disrupting chemicals have adversely affected a variety of fish species. In the vicinity of certain sources (e.g., effluents of water treatment plants) and in the most contaminated areas is this exposure causally linked with the effects on reproductive organs that could have implications for fish populations. However, there is also a more widespread occurrence of endocrine disruption in fish in the U.K., where estrogenic effects have been demonstrated in freshwater systems, in estuaries, and in coastal areas. In mammals, the best evidence comes from the-field studies on Baltic gray and ringed seals, and from the Dutch semifield studies on harbor seals, where both reproduction and immune functions have been impaired by PCBs in the food chain. Reproduction effects resulted in population declines, whereas impaired immune function has likely contributed to the mass mortalities due to morbillivirus infections. Distorted sex organ development and function in alligators has been related to a major pesticide spill into a lake in Florida, U.S.A. The observed estrogenic/antiandrogenic effects in this reptile have been causally linked in experimental studies with alligator eggs to the DDT complex. Although most observed effects currently reported concern heavily polluted areas, endocrine disruption is a potential global problem. This is exemplified by the widespread occurrence of imposex in marine snails and the recent findings of high levels of persistent potential endocrine-disrupting chemicals in several marine mammalian species inhabiting oceanic waters. PMID- 10680770 TI - Changes of the thromboxane A2/prostacyclin balance in the urine of patients with renal diseases analyzed by gas chromatography/selected ion monitoring. AB - The thromboxane A2/prostacyclin (TX/PGI) ratios were measured in patients with renal diseases to elucidate the relationship between the ratios and the pathological changes of the diseases. Urinary stable metabolites of thromboxane A2 and prostacyclin, 11-dehydro-thromboxane B2 and 2,3-dinor-6-keto-prostaglandin F1alpha, respectively, were converted to 1-methyl ester-propylamide-9,12,15-tris dimethylisopropylsilyl ether derivative and 1-methyl ester-6-methoxime-9,12,15 tris-dimethylisopropylsilyl ether derivative, respectively, and applied to a gas chromatography/selected ion monitoring. The TX/PGI ratios of 10 outpatients and 6 inpatients with chronic glomerulonephritis were higher than those of 13 healthy volunteers. In an inpatient with systemic lupus erythematoides, the TX/PGI ratios were gradually lowered to the normal level with the therapies. Furthermore, the ratios seemed to change in advance of the changes of the levels of urinary protein and hematuria. These observations suggested that the TX/PGI ratio was a useful index to assess the pathological condition of renal diseases and the effects of treatment. PMID- 10680771 TI - Neonatal urinary prostanoid excretion. AB - Urinary excretion of prostanoids prostaglandin E2 (PGE2), PGE-M (7alpha-hydroxy 5,11-diketo-2,3,4,5,20-penta-19-carboxyprostano ic acid), 6-keto-PGF1alpha, 2,3 dinor-6-keto-PGF1alpha, thromboxane B2 (TxB2) 2,3-dinor-TxB2 and 11-dehydro-TxB2 was determined by gas chromatography/mass spectrometry in preterm and term infants to show that there is an age-dependent excretion rate of the above prostanoids in infants this young. Group I included premature children with normal postnatal development, Groups II and III included term children who were admitted in the neonatal period for observation because of feeding problems but who were subsequently found to be completely healthy. We present normal data of three primary prostanoids and four prostanoid metabolites. In Group I, excretion rates of 2,3-dinor-TxB2 were significantly lower than in Group II (P = 0.04) and in Group III (P = 0.05). Furthermore, the excretion rate of 11-dehydro-TxB2 in group I was significantly lower than in Group II (P = 0.05). We found no significant age-dependent differences between the three groups in excretion rates of PGE2, PGE-M, 6-keto-PGF1alpha, 2,3-dinor-6-keto-PGF1alpha, and TxB2. PMID- 10680772 TI - Cyclooxygenases-1 and 2 couple to cytosolic but not group IIA phospholipase A2 in COS-1 cells. AB - Phospholipases A2 (PLA2) and cyclooxygenases (COX) are important enzymes responsible for production of potent lipid mediators, including prostaglandins (PG) and thromboxane A2. We investigated coupling between PLA2 and COX isoforms by using transient transfection in COS-1 cells. Untransfected cells, incubated with or without phorbol ester + the Ca2+ ionophore ionomycin, generated trivial amounts of PGE2. In cells co-transfected with cytosolic PLA2 (cPLA2) and COX-1 or COX-2, phorbol ester + ionomycin markedly stimulated PGE2 production. There was no preferential coupling of cPLA2 to either of the COX isoforms. In contrast, group IIA secretory PLA2 (sPLA2) co-transfected with COX-1 or COX-2 did not lead to an increase in PGE2 production, despite high levels of sPLA2 enzymatic activity. Transfection of cPLA2 did not affect basal free arachidonic acid (AA) levels. Phorbol ester + ionomycin stimulated release of AA in cPLA2-transfected COS-1 cells, but not in untransfected cells, whereas sPLA2 transfection (without stimulation) led to high basal free AA. Thus, AA released by cPLA2 is accessible to both COX isoforms for metabolism to PG, whereas AA released by sPLA2 is not metabolized by COX. PMID- 10680773 TI - Suppression of oxytocin-induced prostaglandin F2alpha release after intra-uterine nordihydroguariaretic acid administration in ewes. AB - Intrauterine administration of the 5-lipoxygenase inhibitor nordihydroguariaretic acid (NDGA; 5 mg, bid) on Days 9-14 of the ovine estrous cycle (estrus = Day 0) delayed luteolysis and extended the duration of the estrous cycle (20+/-1, SD, vs. 16+/-1 days; P < 0.01). In control ewes, plasma concentrations of 13,14,dihydro-15-keto prostaglandin F2alpha increased significantly (P < 0.001) following i.v. administration of oxytocin (10 i.u.) on Day 14; in the nordihydroguariaretic acid-treated ewes, however, there was no such increase. In addition, concentrations of endometrial oxytocin receptors were significantly less (P < 0.01) in the nordihydroguariaretic acid-treated ewes (218+/-60 vs. 579+/-66 fmol/mg tissue). These results suggest that 5-lipoxygenase products of arachidonate metabolism may be involved in the control of ovine luteal function. PMID- 10680774 TI - Different actions of noradrenaline and nitric oxide on the output of prostaglandins and progesterone in cultured bovine luteal cells. AB - The effects of noradrenaline (NA) and nitric oxide (NO) on prostaglandins (PGs) and progesterone (P4) secretion during the development of the bovine corpus luteum (CL) were investigated. Bovine luteal cells of early and mid-cycle CL were cultured for 20 to 24 h in medium containing 10% calf serum, washed, and treated with NA or nitrergic agents for an additional 16 h in a serum-free medium. NA (10(-5) M) stimulated P4 from early and mid-cycle CL by 238% and 154% (P < 0.01), respectively. Moreover, although NA induced a twofold increase in PGE2 secretion (P < 0.01) in both examined periods, the effect of NA on PGF2alpha secretion was approximately 1.5 times higher (P < 0.05) in early than in mid-cycle CL. Two NO synthase inhibitors, L-NAME and L-NOARG (both 10(-4) M), stimulated P4 secretion only in mid-luteal cells (P < 0.01), although they did not affect the cells from early CL. Although a NO donor, S-NAP (10(-4) M) inhibited P4 secretion from mid cycle luteal cells (P < 0.05), it strongly stimulated PGE2 in both examined phases (P < 0.001). On the other hand, the output of PGF2alpha was stimulated by S-NAP only in the cells of the mid-cycle CL (P < 0.01). The overall results suggest that adrenergic and nitrergic agents play opposite roles in the regulation of bovine CL functions. Whereas NA may play a supporting role in luteal development, NO may participate in the functional regression of the bovine CL by inhibiting steroidogenesis. PMID- 10680775 TI - No involvement of lipid mediators in a guinea pig model of ultrasonically nebulized distilled water-induced bronchoconstriction. AB - An inhalation of ultrasonically nebulized distilled water (UNDW) induces bronchoconstriction only in asthmatics, but the mechanism underlying the response is not fully understood. We have reported that bronchoconstriction occurs immediately after UNDW is inhaled 20 min after an antigen challenge in guinea pigs. Our aim was to examine the role of lipid mediators in this response. Passively sensitized guinea pigs were anesthetized and artificially ventilated. A sulfidopeptide leukotriene receptor antagonist, KCA-757, and platelet-activating factor antagonists, Y-24180 and E6123, were administered i.v. 15 min after an aerosolized antigen challenge, and UNDW was inhaled 5 min later. KCA-757, Y 24180, or E6123 did not, significantly alter the UNDW-induced bronchoconstriction. Together with our previous data that thromboxane A2 receptor antagonists did not influence the UNDW-induced bronchoconstriction, the present results suggest that lipid mediators are not involved in the UNDW-induced bronchoconstriction in our guinea pig model. PMID- 10680776 TI - Fetal surfactant as a source of arachidonate in human amniotic fluid. AB - The factors responsible for the onset of labor in women are not well understood but it is clear that parturition is associated with increased production of prostanoids and release of arachidonic acid by intrauterine tissues. Pulmonary surfactant is secreted from the fetal lung into the amniotic fluid where its concentration increases toward term. In this paper we have shown that the ability of fetal surfactant to stimulate prostaglandin production by amnion cells is greatly enhanced by pre-incubating surfactant with amniotic fluid. This is due to the release of fatty acids, including arachidonate, from the lipids of fetal surfactant by the sequential action of phospholipase C and diglyceride lipase. Thus, in addition to providing the amnion with a source of arachidonate derived from the intracellular transfer of arachidonate from surfactant phosphatidylcholine to phosphatidylethanolamine and phosphatidylinositol in amnion cells, fetal surfactant also contributes to the pool of free arachidonate in amniotic fluid. PMID- 10680777 TI - The influence of prostaglandins on leukocyte-endothelium interactions in rabbit mesenteric venules. AB - Contradictory results have been reported concerning the effects of prostaglandins (PGs) on leukocyte-endothelium interactions. Therefore, we investigated the in vivo effects of PGE1, PGE2, Iloprost (a stable PGI2-analogue), and also of a combination of these PGs on leukocyte rolling and FMLP-induced leukocyte adhesion in venules of rabbit mesentery. This preparation was used because of its low level of vasoactivity, eliminating hemodynamic effects on leukocyte-endothelium interactions. The mesentery was superfused with PGs or vehicle. After 30 min FMLP was added to the PG-solution for 15 min, whereupon the tissue was superfused with the PG-solution alone for another 30 min. Neither the PGs nor the cocktail influenced leukocyte rolling. During FMLP administration leukocyte adhesion increased and leukocyte rolling decreased; adhesion was highest in the presence of PGE2. The FMLP-induced decrease in leukocyte rolling was similar in all groups. After FMLP administration had been stopped the number of adherent cells almost returned to baseline and the level of leukocyte rolling increased, the baseline level being reached only in the presence of PGE2. In conclusion, these findings indicate that the effects of PGs on leukocyte-endothelium interactions are limited. PMID- 10680779 TI - Seeking genes for MS: big risks for big gains. PMID- 10680778 TI - Tissue prostanoids as biomarkers for chemoprevention of colorectal neoplasia: correlation between prostanoid synthesis and clinical response in familial adenomatous polyposis. AB - Recent studies indicate that sulindac, a nonsteroidal anti-inflammatory drug (NSAID), lowers mucosal prostanoid levels and regresses colorectal adenomas in patients with familial adenomatous polyposis (FAP). To determine whether they are biomarkers for sulindac-mediated chemoprevention of colorectal adenomas, levels of 5 prostanoids [prostaglandin (PG) D2, PGE2, PGF2alpha, thromboxane B2, and 6 keto-PGF1alpha] in the normal-appearing rectal mucosa from 7 FAP patients with a history of subtotal colectomy and ileorectal anastomosis and 4 FAP patients without surgery, were measured in the absence or presence of exogenously added arachidonic acid before the initiation and at the end of 3 months of sulindac treatment. The addition of arachidonic acid resulted in a uniform increase in the levels of all 5 prostanoids although this increase was selectively attenuated in patients with ileorectal anastomosis who took sulindac. In the latter patients, arachidonic acid also augmented the inhibition of prostanoid synthesis by sulindac. In contrast, sulindac failed to attenuate the increase in prostanoid levels resulting from arachidonic acid in patients without previous surgery. Importantly, when measured in the presence of arachidonic acid, the reduction in the levels of all 5 prostanoids due to sulindac was statistically correlated with a reduction in the size and number of adenomas in the two groups of patients combined. These results suggest that tissue prostanoids measured in the presence of arachidonic acid may serve as sensitive and reliable biomarkers in monitoring the clinical responsiveness of FAP patients undergoing chemoprevention for colorectal neoplasia with NSAIDs. PMID- 10680780 TI - Do changes in brain sodium channels cause central pain? PMID- 10680781 TI - Hereditary recurrent focal neuropathies: clinical and molecular features. AB - The authors review the molecular genetics and pathophysiology of hereditary recurrent focal neuropathies: hereditary neuropathy with liability to pressure palsies (HNPP) and hereditary neuralgic amyotrophy (HNA). Significant progress in the understanding of HNPP and HNA has been achieved. HNPP and HNA are distinct clinical and pathologic disease entities with autosomal dominant inheritance. Molecular genetic studies have shown that HNPP and HNA are located on chromosome 17 but at distinct genetic loci (17p11.2 for HNPP, 17q25 for HNA). The 1.5 megabase deletion in 17p11.2 is the major cause of HNPP. This interstitial deletion causes the complete loss of one allele of the peripheral myelin protein 22 (PMP22) gene. Interestingly, rare HNPP patients are found without the 1.5 megabase deletion. However, these patients have distinct mutations in the PMP22 gene resulting in altered expression of the PMP22 protein. Current molecular genetic tests and clinical guidelines allow improved diagnosis, prognosis, and genetic counseling for patients with HNPP. Such tests are not available for HNA, because the disease-causing gene remains unknown. Molecular genetic advances in HNPP and HNA, as well as the study of transgenic animal and cellular models, will provide a more precise understanding of the disease mechanisms and will lead to the development of effective therapeutic tools for patients with inherited and sporadic recurrent peripheral neuropathies. PMID- 10680782 TI - Glutathione S-transferase polymorphisms in MS: their relationship to disability. AB - BACKGROUND: Oxidative stress has been implicated in inflammatory demyelination. The glutathione S-transferase (GST) supergene family encodes isoenzymes that appear to be critical in protection against oxidative stress. Certain GST loci are polymorphic, demonstrating alleles that are null (GSTM1/GSTT1), encode low activity variants (GSTP1), or are associated with variable inducibility (GSTM3). OBJECTIVES: To investigate the association between clinical outcome in MS and allelic variants of GSTM1, GSTM3, GSTT1, and GSTP1. METHODS: Four hundred patients with clinically definite MS were studied. Disability was measured using the Kurtzke Expanded Disability Status Scale (EDSS). Disability was graded as mild (EDSS 0-4), moderate (4.5-5.5), or severe (EDSS 6-10). PCR-based genotyping was performed using DNA extracted from lymphocytes. Significant associations between GST genotypes and clinical outcome were corrected for gender, onset age, and disease duration using logistic regression. RESULTS: We found that the GSTM3 AA genotype was associated with severe disability in patients with a disease duration of more than 10 years (p = 0.027, n = 177, OR = 2.4, 95% CI = 1.1-5.0). Homozygosity for both GSTM1*0 and GSTP1*Ile105 containing allele was associated with severe disability in patients with a disease duration greater than 10 years (p = 0.022, n = 179, OR = 5.0, 95% CI = 1.3-19.8). CONCLUSIONS: Our results suggest that long-term prognosis in MS is influenced by a genetically determined ability to remove the toxic products of oxidative stress. PMID- 10680783 TI - Cortical cerebral metabolism correlates with MRI lesion load and cognitive dysfunction in MS. AB - OBJECTIVE: To study the association between the cortical cerebral metabolic rate of glucose (CMRglc), MRI T2-weighted total lesion area (TLA), cognitive dysfunction, and neurologic disability in MS. BACKGROUND: MRI lesion load is widely used in the clinical evaluation of the MS patient but little is known about the associated changes in cortical activation. METHODS: Twenty-three patients with clinically definite MS underwent measurements of CMRglc, TLA, motor evoked potentials (MEPs), and cognitive and neurologic disability. CMRglc was calculated using PET and 18-F-deoxyglucose and compared with nine normal control subjects. RESULTS: Reductions in CMRglc (p < 0.01) were found in the cortical global and regional lobar measurements. Furthermore, regional CMRglc (rCMRglc) was reduced in the dorsolateral prefrontal cortex, orbitofrontal cortex, caudate, putamen, thalamus, and hippocampus. Global cortical CMRglc correlated with TLA (Spearman rank correlation coefficient [SRCC] = -0.66, p = 0.001), and rCMRglc correlated with regional lesion load in all cerebral lobes (p < or = 0.05). Global cortical CMRglc and cognitive disability also correlated (SRCC = 0.58, p = 0.015), and stepwise regression analysis showed a significant association between rCMRglc of the right thalamus and cognitive performance as well as TLA. There was no correlation between CMRglc and neurologic disability (Expanded Disability Status Scale) or MEP. CONCLUSION: Global and regional cortical CMRglc is reduced significantly in MS patients compared with normal control subjects. Furthermore, the CMRglc reductions correlate with TLA as well as with cognitive dysfunction, which indicates that MRI white matter lesion burden has a deteriorating effect on cortical cerebral neural function. PMID- 10680784 TI - Intravenous lidocaine in central pain: a double-blind, placebo-controlled, psychophysical study. AB - OBJECTIVE: To investigate the effects of systemic administration of lidocaine on different components of neuropathic central pains by quantitative sensory testing. METHODS: The efficacy of systemic lidocaine (5 mg/kg IV over 30 minutes) was evaluated in a double-blind, placebo-controlled, and cross-over fashion, on both spontaneous ongoing pain and evoked pains (allodynia and hyperalgesia) in 16 patients with chronic poststroke (n = 6) or spinal cord injury (n = 10) related pain. RESULTS: Lidocaine was significantly superior to the placebo (saline) in reducing the intensity of spontaneous ongoing pain for up to 45 minutes after the injection: 10 of 16 patients (62.5%) receiving lidocaine showed a significant reduction in spontaneous pain, whereas only six patients showed this after the placebo. Lidocaine also significantly reduced the intensity of brush-induced allodynia and mechanical hyperalgesia, but was no better than the placebo against thermal allodynia and hyperalgesia. In general, the side effects were moderate and consisted mainly of lightheadedness (44%). CONCLUSIONS: Systemic lidocaine can induce a significant and selective reduction of several components of pain caused by CNS injuries. The observed preferential antihyperalgesic and antiallodynic effects of this drug suggest a selective central action on the mechanisms underlying these evoked pains. PMID- 10680785 TI - Fear recognition deficits after focal brain damage: a cautionary note. AB - OBJECTIVE: To test the hypothesis that fear recognition deficits in neurologic patients reflect damage to an emotion-specific neural network. BACKGROUND: Previous studies have suggested that the perception of fear in facial expressions is mediated by a specialized neural system that includes the amygdala and certain posterior right-hemisphere cortical regions. However, the neuropsychological findings in patients with amygdala damage are inconclusive, and the contribution of distinct cortical regions to fear perception has only been examined in one study. METHODS: We studied the recognition of six basic facial expressions by asking subjects to match these emotions with the appropriate verbal labels. RESULTS: Both normal control subjects (n = 80) and patients with focal brain damage (n = 63) performed significantly worse in recognizing fear than in recognizing any other facial emotion, with errors consisting primarily of mistaking fear for surprise. Although patients were impaired relative to control subjects in recognizing fear, we could not obtain convincing evidence that left, right, or bilateral lesions were associated with disproportionate impairments of fear perception once we adjusted for differences in overall recognition performance for the other five facial emotion categories. The proposed special role of the amygdala and posterior right-hemisphere cortical regions in fear perception was also not supported. CONCLUSIONS: Fear recognition deficits in neurologic patients may be attributable to task difficulty factors rather than damage to putative neural systems dedicated to fear perception. PMID- 10680786 TI - Memory and MRI-based hippocampal volumes in aging and AD. AB - OBJECTIVE: To demonstrate structural-functional relationships between MRI-based volumetric measurements of medial temporal lobe structures and cognitive function. BACKGROUND: Previous work has documented the ability of MRI-based measurements of the hippocampus to discriminate between age-matched control subjects and patients with very mild AD. Relatively less is known about the correlation between medial temporal lobe structures and cognitive functions. METHOD: We evaluated structural-functional relationships among the hippocampal formation, parahippocampal gyrus, and amygdala, and measures of memory, language, and general cognitive performance in 220 probable AD patients and normal control subjects. Standardized instruments of memory and general cognitive function were used to assess subjects enrolled in a longitudinal study of aging and dementia. RESULTS: The volume of the hippocampal formation predicted performance on most acquisition and recall measures across the spectrum of normal aging and AD. If the groups were segregated, most of the expected associations between medial temporal lobe structures and memory measures were observed in the AD patients. CONCLUSION: MRI-based hippocampal volumetry accurately depicts the structural functional relationships between memory loss and hippocampal damage across the spectrum from normal aging to dementia. PMID- 10680787 TI - A randomized controlled trial of prednisone in Alzheimer's disease. Alzheimer's Disease Cooperative Study. AB - BACKGROUND: Laboratory and epidemiologic studies suggest that anti inflammatory/immunosuppressive therapy may be useful in the treatment of AD. In preliminary studies, a regimen of low to moderate dose prednisone was found to suppress peripheral inflammatory markers without adverse effects in subjects with AD. METHODS: We conducted a randomized, placebo-controlled multicenter trial to determine whether prednisone treatment slowed the rate of cognitive decline in AD. The active treatment regimen consisted of an initial dose of 20 mg of prednisone daily for 4 weeks tapered to a maintenance dose of 10 mg daily for 1 year, followed by gradual withdrawal during an additional 16 weeks. The primary outcome measure was the 1-year change in the cognitive subscale of the AD Assessment Scale. RESULTS: A total of 138 subjects were randomized to the drug and placebo groups. There was no difference in cognitive decline between the prednisone and placebo treatment groups in the primary intent-to-treat analysis, or in a secondary analysis considering completers only. Subjects treated with prednisone showed behavioral decline compared with those in the placebo group. CONCLUSION: A low-dose regimen of prednisone is not useful in the treatment of AD. PMID- 10680788 TI - APOE and AD concordance in twin pairs as predictors of AD in first-degree relatives. AB - OBJECTIVE: To examine the independent effects of the APOE genotype (APOE) and concordance for AD in twin pairs on the occurrence of AD in first-degree relatives. BACKGROUND: Studies of twins have been undertaken to investigate the influence of genes in a variety of conditions, including AD. A previous study, performed before reports linking APOE to AD, demonstrated an increase in AD among first-degree relatives of twins concordant for AD compared with relatives of discordant twins. METHODS: In a sample of 94 twin pairs the authors examined the association between concordance for AD within the twin pair and family history of AD among first-degree relatives of twins. They then examined the extent to which the presence of the APOE epsilon4 allele in the twin pair explains the association between concordance for AD within the twin pair and family history of AD. RESULTS: Concordance among twins was associated with increased risk of AD among relatives (logrank test, chi2 = 12.558; p = 0.0004), and the presence of at least one APOE epsilon4 allele in each member of the twin pair is also associated with increased risk of AD among family members (logrank test, chi2 = 7.712; p = 0.0055). CONCLUSIONS: APOE genotype explains much but not all of the association between concordance among twins and increased familial risk of AD. PMID- 10680789 TI - Associations between circulating sex steroid hormones and cognition in normal elderly women. AB - OBJECTIVE: To provide exploratory analyses of associations between levels of several sex hormones and cognitive performance in elderly women. BACKGROUND: Sex steroid hormones are implicated in the cognitive processes of the adult brain. Comparing cognitive performance across or between conditions associated with different hormone levels, such as phases of the menstrual cycle, surgical menopause, and estrogen replacement therapy suggests conditions with higher levels of estrogen are associated with better verbal memory and possibly worse visuospatial ability. METHOD: The authors measured circulating sex hormone levels in 39 highly educated, nondemented, predominantly white elderly women. Levels were correlated with neuropsychological performance, controlling for age, education, frequency of prior testing, use of estrogen replacement, and depression. RESULTS: High estradiol levels were associated with better delayed verbal memory and retrieval efficiency, whereas low levels were associated with better immediate and delayed visual memory. Levels of testosterone were related positively to verbal fluency. Levels of progesterone and androstenedione were unrelated to cognitive performance. CONCLUSIONS: Both estrogen and testosterone showed associations with cognitive performance. Estrogen may enhance, and depress, specific cognitive skills. PMID- 10680790 TI - A randomized trial of 3,4-diaminopyridine in Lambert-Eaton myasthenic syndrome. AB - OBJECTIVES: The authors report the results of a prospective, placebo-controlled, randomized study to evaluate the effectiveness of 3,4-diaminopyridine (DAP) in patients with Lambert-Eaton myasthenic syndrome (LEMS) and to determine the acute and long-term side effects of DAP. METHODS: Twenty-six patients with LEMS completed a two-arm parallel treatment protocol in which DAP, 20 mg three times daily, or placebo was given blindly for 6 days, and a quantitative examination of muscle strength (the quantitative myasthenia gravis [QMG] score) was used as the primary measure of efficacy. After the blinded study, patients were given open label DAP and monitored for side effects as long as there was symptomatic improvement. RESULTS: Twelve patients took DAP, and 14 took placebo. There was no difference in the age of LEMS onset, gender distribution, incidence of lung cancer, or baseline muscle strength between the patients who were randomly assigned to receive placebo and those randomly assigned to DAP. Statistical analysis using the Wilcoxon's rank sum test demonstrated that patients who received DAP had a significantly greater improvement in the QMG score and in the summated amplitude of compound muscle action potentials recorded from three sentinel limb muscles. All but one LEMS patient had significant symptomatic improvement from subsequent open-label DAP. Side effects of DAP were negligible, consisting of perioral and digital paresthesia. Laboratory measurements demonstrated no evidence of toxicity affecting liver, renal, hematologic, endocrinologic, encephalographic, or electrocardiologic function acutely or after 6 months of open-label DAP. CONCLUSIONS: This study corroborates previous studies and many years of clinical experience showing that DAP is an effective and safe treatment for LEMS. PMID- 10680791 TI - GCG genetic expansions in Italian patients with oculopharyngeal muscular dystrophy. AB - OBJECTIVE: To screen Italian patients with oculopharyngeal muscular dystrophy (OPMD) for GCG repeat expansions in the Poly(A) binding-protein 2 (PABP2) gene. BACKGROUND: Oculopharyngeal muscular dystrophy is an adult-onset autosomal dominant muscle disease linked to 14q11 pathologically characterized by unique 8.5 nm intranuclear filaments in skeletal muscle fibers. Short expansions of a (GCG)6 repeat located in exon 1 of the newly isolated PABP2 gene have been demonstrated in a large number of OPMD families. METHODS: We studied 18 patients diagnosed with OPMD. A muscle biopsy was performed in 16 patients. Screening for the pathologic expansion was performed on a PCR amplified DNA fragment encompassing the GCG repeat. RESULTS: Heterozygous (GCG)-repeat expansions were detected in 13 patients in association with (GCG)6 normal allele or (GCG)7 polymorphic allele. All the patients whose muscle biopsy showed typical 8.5 nm intranuclear filaments had a mutated PABP2 allele. Five patients with no intranuclear filaments were homozygous for the normal (GCG)6 allele. The pathologic expansion appeared to be stable with no variation among family members and between different tissues as blood and skeletal muscle in the same individual. CONCLUSIONS: These data 1) further confirm PABP2 gene analysis as a valuable tool in OPMD diagnosis; 2) indicate that PABP2 gene mutations are always present among Italian patients with morphologically proven OPMD, suggesting genetic homogeneity of the disease; and 3) strengthen the putative role of mutated PABP2 protein in filamentous inclusions accumulation. PMID- 10680792 TI - Distal acquired demyelinating symmetric neuropathy. AB - OBJECTIVE: To characterize an acquired, symmetric, demyelinating neuropathic variant with distal sensory or sensorimotor features. BACKGROUND: Classic chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) patients have prominent proximal and distal weakness. However, chronic demyelinating neuropathies may present with different phenotypes. An approach that distinguishes these disorders primarily according to the pattern of weakness may be useful to the clinician. METHODS: A total of 53 patients with acquired symmetric demyelinating polyneuropathies were classified primarily according to the pattern of the neuropathy and secondarily according to the presence and type of monoclonal protein (M-protein) in this retrospective review. The authors distinguished between patients with distal sensory or sensorimotor involvement, designated as distal acquired demyelinating symmetric (DADS) neuropathy, from those with proximal and distal weakness, who were designated as CIDP. RESULTS: M-proteins were present in 22% of patients with CIDP. There were no features that distinguished clearly between CIDP patients with or without an M-protein, and nearly all of these patients responded to immunomodulating therapy. In contrast, nearly two-thirds of the patients with DADS neuropathy had immunoglobulin M (IgM) kappa monoclonal gammopathies, and this specific combination predicted a poor response to immunomodulating therapy. Antimyelin-associated glycoprotein (anti MAG) antibodies were present in 67% of these patients. CONCLUSION: Distinguishing acquired demyelinating neuropathies by phenotype can often predict the presence of IgM kappa M-proteins, anti-MAG antibodies, and responses to immunomodulating therapy. PMID- 10680793 TI - Mild forms of Guillain-Barre syndrome in an epidemiologic survey in The Netherlands. AB - OBJECTIVE: Assessment of incidence rates of Guillain-Barre syndrome (GBS) in the Netherlands over a 10-year period; investigation of a relationship between possible seasonality in GBS and the occurrence of preceding infections; and determination of distinctive characteristics in patients with GBS who are only mildly affected (able to walk unaided at nadir). METHOD: Records of patients with GBS admitted between 1987 and 1996 from all 45 hospitals in the southwest Netherlands were evaluated, covering a population of 4.2 million inhabitants. RESULTS: A total of 476 patients met National Institute for Neurological and Communicative Disorders and Stroke criteria for GBS. This resulted in a crude incidence rate (IR) of 1.18/100,000 inhabitants. This IR increased linearly with age (p < 0.001). Men were more frequently affected than women (p < 0.001). No seasonal preponderance for GBS, nor for any of the preceding infections, was found. Patients under 50 years of age (p < 0.001) and men (p = 0.01) were more frequently found in the mildly affected group. In both groups a preceding infection was reported in 70% of the cases. In the severely affected group, serologic evidence for infection with Campylobacter jejuni, cytomegalovirus, Epstein-Barr virus, or Mycoplasma pneumoniae was found more frequently than in the mildly affected group (41% versus 16%, p = 0.001). CONCLUSIONS: Overall IR in the Netherlands are similar to those found in other studies. The incidence increases linearly with age and men are more frequently affected than women. Distinctive characteristics for mildly and severely affected patients were found regarding age, sex, and preceding infections. This suggests that other infectious agents or host factors may be involved in mild forms of GBS. PMID- 10680794 TI - Driving in adults with refractory localization-related epilepsy. Multi-Center Study of Epilepsy Surgery. AB - OBJECTIVE: To examine the frequency of driving an automobile and characteristics associated with driving in individuals with refractory localization-related epilepsy. BACKGROUND: Driving is generally restricted and monitored in people with epilepsy. Little is known about the frequency of driving and subsequent accidents specifically in individuals with uncontrolled epilepsy. METHODS: In an ongoing, prospective, multicenter study of resective epilepsy surgery, individuals were interviewed when they presented for surgical evaluation. Analyses were conducted using chi-square, t-tests, and multiple logistic regression. RESULTS: Of 367 eligible participants, 115 (31.3%) had driven in the last year, most on at least a weekly basis. In a multivariable analysis, factors associated with an increased likelihood of driving were having a current license (OR = 10.71, p < 0.001) and ever having had a license (OR = 3.86, p = 0.003). Younger individuals were also more likely to drive. Lower levels of driving were found in women (OR = 0.31, p < 0.001), individuals who were self-described as disabled (OR = 0.20, p < 0.001), and those who were employed full-time (OR = 0.43, p = 0.03) or part-time (OR = 0.15, p = 0.005). At some point in the past, 144 individuals experienced one or more seizures while driving, and 98 experienced at least one accident because of a seizure. Of those who had accidents, 94% reported property damage, 32% had an injury, and 20% caused injury to others. CONCLUSION: Despite restrictions, almost one third of individuals with refractory epilepsy drive. Understanding why they do may help identify means of modifying this behavior or identifying services that, if provided, would help people with uncontrolled epilepsy forego driving. PMID- 10680795 TI - Seizure outcome after temporal lobectomy for temporal lobe epilepsy: a Kaplan Meier survival analysis. AB - OBJECTIVE: To determine seizure outcome and its predictors in patients with medically refractory temporal lobe epilepsy (TLE) after temporal lobectomy (TL). BACKGROUND: TL is the most common surgical procedure performed in adolescents and adults for the treatment of medically refractory TLE. Seizure outcome has been reported extensively during the first few postoperative years, but little is known beyond that time. METHODS: The authors analyzed seizure outcome in 79 patients who underwent TL for epilepsy at the Duke University Medical Center from 1962 through 1984. Patients with less than 2 years of follow-up and degenerative disorders were excluded. Predictors of seizure outcome were analyzed using Kaplan Meier survival analyses. RESULTS: The mean follow-up was 14 years (range, 2.1 to 33.6 years). Using Engel's classification, 65% of patients were class I, 15% were class II, 11% were class III, and 9% were class IV. At least one postoperative seizure occurred in 55% of subjects. The majority of recurrences (86%) took place within 2 years of surgery. Later recurrences tended not to lead to medical intractability. Higher monthly preoperative seizure frequency was associated with poor seizure outcome. A seizure-free state at 2 years was found to be a better predictor of long-term outcome than the 6-, 12-, and 18-month landmarks. CONCLUSIONS: TL provides sustained, long-term benefit in patients with medically refractory TLE. Seizure-free status at 2 years from the time of surgery is predictive of long-term remission. PMID- 10680796 TI - An EEG should not be obtained routinely after first unprovoked seizure in childhood. AB - OBJECTIVE: To quantify and analyze the value of expected information from an EEG after first unprovoked seizure in childhood. BACKGROUND: An EEG is often recommended as part of the standard diagnostic evaluation after first seizure. METHODS: A MEDLINE search from 1980 to 1998 was performed. From eligible studies, data on EEG results and seizure recurrence risk in children were abstracted, and sensitivity, specificity, and positive and negative predictive values of EEG in predicting recurrence were calculated. Linear information theory was used to quantify and compare the expected information from the EEG in all studies. Standard test-treat decision analysis with a treatment threshold at 80% recurrence risk was used to determine the range of pretest recurrence probabilities over which testing affects treatment decisions. RESULTS: Four studies involving 831 children were eligible for analysis. At best, the EEG had a sensitivity of 61%, a specificity of 71%, and an expected information of 0.16 out of a possible 0.50. The pretest probability of recurrence was less than the lower limit of the range for rational testing in all studies. CONCLUSIONS: In this analysis, the quantity of expected information from the EEG was too low to affect treatment recommendations in most patients. EEG should be ordered selectively, not routinely, after first unprovoked seizure in childhood. PMID- 10680797 TI - Predictors of outcome in pediatric epilepsy surgery. AB - OBJECTIVE: To determine the correlation between pre- and perioperative variables on the outcome of children undergoing focal resections for medically intractable partial epilepsy. METHODS: Retrospective analysis of pre- and perioperative variables in a cohort of 75 patients younger than 12 years of age who underwent excisional surgery and had at least 1 year of follow-up. Outcome, measured by postoperative seizure frequency, was analyzed as a function of age at seizure onset, duration of epilepsy, presence of cognitive impairment, lobe of seizure origin, presence of a lesion, histopathology, and completeness of resection. Completeness of resection was defined on the basis of excising both the entire structural lesion if present and the region revealing prominent interictal and ictal abnormalities on intracranial EEG. RESULTS: Seventy-seven percent of patients had good outcomes (class 1 or 2), and 59% were seizure-free. Lesional status, site of resection, and pathologic diagnoses were not significant predictors of outcome except for in multilobar resection, for which overall outcome was relatively poor (44% class 3 or 4; 22% seizure-free). Completeness of resection was the only significant predictor of good outcome (p < 0.001), with 92% of patients who underwent complete resection of the epileptogenic zone achieving good outcome compared with 50% of patients who had incomplete resections. CONCLUSION: In this series of pediatric patients, complete resection of the lesion and the electrographically abnormal region was the main determinant of outcome after focal resections. Except for multilobar resections, other factors examined in this study did not significantly influence postoperative seizure prognosis and should not influence candidate selection for the surgical process. PMID- 10680798 TI - Reduction of corpus callosum growth after severe traumatic brain injury in children. AB - OBJECTIVE: To study effects of closed head injury (CHI) severity on development of corpus callosum (CC) in children, using MRI. BACKGROUND: Vulnerability of CC to diffuse axonal injury has been shown in adults and children by neuropathologic and MRI studies. Given continued development of CC through the second decade, serial MRI could characterize effects of CHI on CC growth in children. METHOD: MRI performed at 3 and 36 months after severe (mean age = 10.3 years, n = 25) and mild to moderate (mean age = 9.7 years, n = 28) CHI. Mild to moderate and severe CHI groups did not differ in demographic features. Morphometry of T1-weighted midsagittal CC by two operators with satisfactory interrater reliability yielded uncorrected and corrected CC volume. RESULTS: An interaction of occasion with CHI severity was present as CC area decreased from 3 to 36 months in severely injured children and increased in the mild to moderate CHI group. Uncorrected CC area was correlated with acute CHI severity and functional outcome at 36 months postinjury. CONCLUSIONS: Morphometric measurement of CC area provides a useful index of diffuse injury, which is related to functional outcome of CHI in children. PMID- 10680799 TI - Hypersomnia after head-neck trauma: a medicolegal dilemma. AB - OBJECTIVES: To evaluate the severity of daytime sleepiness in patients with a history of head trauma who complain of daytime somnolence, to investigate polygraphic abnormalities during nocturnal sleep, and to determine whether daytime sleepiness was the cause or consequence of the head trauma. METHODS: The authors performed a systematic evaluation of 184 patients comprised of clinical interviews, sleep disorders questionnaires, sleepiness and depression scales, medical and neurologic evaluations, sleep logs with actigraphy, nocturnal polysomnography, and the Multiple Sleep Latency Test (MSLT). Assessments of sleepiness before the accident were based on bed partner interviews, coworker and employer reports, health reports, driving records, and employment history that included absenteeism. RESULTS: Post-traumatic complaint of somnolence was associated with variable degrees of impaired daytime functioning in more than 98% of patients. Patients who were in a coma for 24 hours, who had a head fracture, or who had immediate neurosurgical interventions were likely to have scores > 16 points on the Epworth Sleepiness Scale (ESS) and < or = 5 minutes on the MSLT. Pain at night was an important factor in nocturnal sleep disruption and daytime sleepiness. Sleep-disordered breathing was a common finding and was the only finding in whiplash patients with daytime sleepiness. Extensive evaluation of pretrauma behavior supported the conclusion that the onset of symptomatic sleep disordered breathing was associated with the trauma. The patients who showed a "compulsive presleep behavior" were severely impaired in performing their daily activities. CONCLUSIONS: A systematic approach is required when dealing with patients complaining of hypersomnia following a head-neck trauma. PMID- 10680800 TI - Risk factors and outcome of patients with carotid artery stenosis presenting with lacunar stroke. North American Symptomatic Carotid Endarterectomy Trial Group. AB - OBJECTIVE: To examine the relationship between carotid artery stenosis, other risk factors, and lacunar stroke. BACKGROUND: Carotid artery stenosis in patients presenting with lacune stroke may be coincidental or causal. The distinction by risk factor profile is uncertain. The risk and cause of subsequent stroke, and benefit of carotid endarterectomy (CE) is unknown. METHODS: Stroke in patients entering the North American Symptomatic Carotid Endarterectomy Trial were classified as nonlacunar, possible lacune (symptoms without CT lacunae), or probable lacune (symptoms with CT lacunae). RESULTS: Of 1,158 patients with hemispheric stroke, 493 had features of lacunar stroke (283 possible and 210 probable). Lacunar stroke presented more commonly in patients with milder (<50%) degrees of internal carotid artery (ICA) stenosis (p = 0.003). History of diabetes and hyperlipidemia, not hypertension, were associated independently even after accounting for the degree of stenosis. Medically treated patients presenting with nonlacunar stroke had a low risk of subsequent lacunar events of 2.9% at 3 years in comparison with 9.2% for probable lacunar presentation (p = 0.03). For patients with 50 to 99% ICA stenosis, the relative risk reductions (RRRs) in stroke from CE were 35% when the presenting stroke was probable lacunar versus 61% when the stroke was nonlacunar. Patients presenting with a possible lacunar stroke had a 53% RRR. CONCLUSIONS: History of diabetes and hyperlipidemia were more important than arterial hypertension as risk factors for patients with lacunar stroke. Patients presenting with lacunar stroke more often had milder ICA stenosis. Although CE reduced the risk of stroke in all patients with 50 to 99% ICA stenosis, lesser benefits were observed in patients presenting with lacunar stroke. PMID- 10680801 TI - rtPA intravenous thrombolysis in anterior choroidal artery territory stroke. AB - OBJECTIVE: To study the possible specific response to recombinant tissue plasminogen activator (rtPA) thrombolysis of anterior choroidal artery (AChA) stroke. BACKGROUND: Outcome and response after rtPA thrombolysis are possibly better in small-vessel infarcts, but a specific study of AChA stroke has not yet been performed. METHODS: The authors proposed an open trial of IV rtPA within 7 hours in patients age 20 and 81 years with all types of internal carotid artery territory stroke if the baseline Scandinavian Stroke Scale (SSS) score was less than 48. A dose of rtPA 0.8 mg/kg was infused over 90 minutes. Of 114 consecutive patients, 9 patients (7.9%) exhibited hypodensity in the AChA territory on day 1 brain CT. RESULTS: Seven of nine patients with AChA infarct had a primary early recovery within 6 hours after the initiation of rtPA infusion. In addition, recovery was complete in five patients and partial in two patients. No intracerebral hematoma was observed. Three patients had a "reinfarct syndrome" at 12, 25, and 48 hours respectively. However, in the two latter patients treated with IV heparin, the deficit disappeared again with the increase of heparin dose in one patient and disappeared spontaneously in the other patient. The overall outcome at day 90 was six total recoveries in nine patients (66%). Patients with a final good outcome had a slight "unstructured" hypodensity in the AChA territory on day 1 brain CT, whereas patients with a bad outcome had the classic "structured" hypodensity of AChA territory stroke. CONCLUSION: These data support a specific quick response of AChA territory stroke to IV rtPA thrombolysis, probably due to the small size of the artery and of the "clot." The high frequency of the reinfarct syndrome is a clinical fact that is difficult to explain. Efficient heparin treatment after 24 hours may control the reinfarct syndrome in some patients. PMID- 10680802 TI - Multiple acute stroke syndrome: marker of embolic disease? AB - OBJECTIVE: To determine the frequency and etiologic significance of multiple acute ischemic lesions in stroke. BACKGROUND: Although patients may have more than one stroke during the course of their lives, acute ischemic stroke is usually thought of as a single event. Using diffusion-weighted imaging (DWI), an MRI technique that detects ischemic injury within minutes after onset, we have often observed multiple acute ischemic lesions. METHODS: The MRI scans of 59 consecutively studied patients were reviewed to determine the frequency and etiologic significance of multiple acute ischemic lesions on DWI. RESULTS: Multiple acute ischemic lesions were present in 10 (17%) of 59 patients. The lesions usually occurred within one major circulation (anterior or posterior), but in two patients (3%), lesions occurred in both cerebral hemispheres or in the anterior and the posterior circulations. The lesions often were small and resulted from presumed multiple emboli or the break-up of an embolus. Two patients had internal carotid artery occlusive disease and four had a cardiac or aortic source. In the other four patients the source was not determined. Lesions larger than 1 cm in diameter progressed to infarction, but some smaller lesions were not seen on follow-up T2-weighted imaging. CONCLUSIONS: Multiple acute stroke lesions on DWI are common and could be caused by multiple emboli or the breakup of an embolus. In some cases it might become possible to make early inferences concerning the stroke mechanism that could be of use for immediately directing the clinical work-up and treatment of the patient. PMID- 10680803 TI - Effectiveness of t-PA in acute ischemic stroke: outcome relates to appropriateness. AB - OBJECTIVE: To examine whether the demonstrated efficacy of tissue-type plasminogen activator (t-PA) for acute ischemic stroke can be effective in a community setting. METHODS: Sixty-eight consecutive patients with acute ischemic stroke treated with IV t-PA within 3 hours of symptom onset by attending general neurologists in a busy teaching hospital. Outcome measures at 3 months were the National Institute of Health Stroke Scale (NIHSS), functional outcome (independence [modified Rankin score 0-2], dependence [modified Rankin score 3 5], and death), and symptomatic hemorrhage. Appropriately treated patients were defined by adherence to the National Institute of Neurological Disorders and Stroke (NINDS) guidelines. Effectiveness is expressed as the absolute risk reduction in which the baseline risk is assumed to be similar to that of the NINDS control group. RESULTS: Of 68 consecutively treated patients (with a mean baseline NIHSS score of 15 +/- 6), 26 (38%) made a full recovery and 39 (57%) made an independent recovery. The 11 patients who violated protocol had a lower probability of independence (p < 0.02) and full neurologic recovery (p < 0.02) and a higher probability of symptomatic hemorrhage (p < 0.05) and death (p < 0.01) compared with those of 57 patients treated according to NINDS guidelines. CONCLUSIONS: The use of t-PA for stroke in this community is effective with a number needed to treat of six. The risk of symptomatic hemorrhage is similar to that noted in randomized trials. Treating patients who violate protocol results in excess risk with no observable benefit. PMID- 10680804 TI - Identification of lacunar infarcts before thrombolysis in the ECASS I study. AB - BACKGROUND: The identification of lacunar infarcts before thrombolysis would make it possible either to exclude them from treatment or to show that they also may benefit from it. OBJECTIVE: To determine whether clinical presentation or early CT findings of patients enrolled in the first European Cooperative Acute Stroke Study (ECASS I) trial would identify lacunar infarcts before treatment. METHODS: Predictive values, sensitivity, specificity, and accuracy of clinical presentation as pure motor hemiparesis (PMH) or sensorimotor stroke (SMS) syndromes and of baseline CT findings in predicting lacunar infarcts were calculated in the ECASS I patients. RESULTS: Of 514 patients, 44 placebo (17%) and 44 recombinant tissue plasminogen activator (rt-PA) (18%) patients had PMH/SMS involving at least two of three areas. Thirty-one placebo (12%) and 32 rt PA (13%) patients had PMH/SMS involving three areas. The 7-day CT was compatible with a lacunar infarct in 32 placebo (12%) and 44 rt-PA (18%) patients. PMH/SMS involving at least two areas had a positive predictive value of 30% both in placebo and rt-PA patients, whereas positive predictive values of the involvement of three areas were 23% and 31%. Those of absence of early CT signs were 21% and 30%, and those of leukoaraiosis or previous lacunar infarcts were 21% and 23%. Positive predictive values of PMH/SMS involving at least two areas combined with absence of early CT signs were 36% in placebo and 33% in t-PA patients, and those of PMH/SMS plus leukoaraiosis or previous lacunes were 28% and 7%, respectively. CONCLUSIONS: In the ECASS I trial, lacunar infarcts were not recognizable on clinical grounds, and early CT findings, alone or in combination with the clinical picture, added poorly to the differential diagnosis. PMID- 10680805 TI - Temporal evolution of ischemic injury evaluated with diffusion-, perfusion-, and T2-weighted MRI. AB - OBJECTIVE: Ischemic lesions seen on diffusion-weighted imaging (DWI) are reversible if reperfusion is performed within minutes after the onset of ischemia. This study was designed to determine whether acute reversibility of DWI abnormalities is transient following brief temporary focal brain ischemia and to characterize the temporal evolution of in vivo ischemic lesions. METHODS: Eight rats were subjected to 30 minutes of temporary middle cerebral artery occlusion and underwent diffusion-, perfusion-, and T2-weighted MRI during occlusion; immediately after reperfusion; 30, 60, and 90 minutes after reperfusion; and 12, 24, 48, and 72 hours after reperfusion. Average apparent diffusion coefficient (ADCav) values and the cerebral blood flow index (CBFi) ratio were calculated in both the lateral caudoputamen and overlying cortex at each time point. The size of the in vivo ischemic abnormalities was calculated from the ADCav and the T2 maps. Postmortem triphenyltetrazolium chloride (TTC) staining was used to verify ischemic injury. RESULTS: Both the CBFi ratio and ADCav values declined significantly in the two regions during occlusion. The CBFi ratio recovered immediately after reperfusion and remained unchanged over 72 hours. However, ADCav values returned to normal at 60 to 90 minutes and secondarily decreased at 12 hours after reperfusion as compared with those in the contralateral hemisphere. The extent of the in vivo ischemic lesions maximized at 48 hours and was highly correlated with TTC-derived lesion size. CONCLUSIONS: Acute recovery of initial ADCav-defined lesions after reperfusion is transient, and secondary ADCav-defined lesions develop in a slow and delayed fashion. PMID- 10680806 TI - Differentiation of atypical parkinsonian syndromes with routine MRI. AB - OBJECTIVE: To evaluate the use of routine MRI in differentiating between patients with progressive supranuclear palsy (PSP), multiple system atrophy (MSA), corticobasal degeneration (CBD) and control subjects. METHODS: Two neuroradiologists rated blindly and independently axial T2-weighted and proton density MR images of 54 patients with MSA, 35 patients with PSP, 5 patients with CBD, and 44 control subjects. RESULTS: More than 70% of patients with PSP and more than 80% of patients with cerebellar predominant MSA could be classified correctly with 0.5-T or 1.5-T scans, and no patient in these groups was misclassified. In the remaining patients an unequivocal differentiation could not be made. However, only approximately 50% of patients with parkinsonism predominant MSA could be classified correctly, and 19% of them (all of whom had had 0.5-T scans) were misclassified. CONCLUSIONS: Characteristic findings on routine MRI, either 1.5 T or 0.5 T, can contribute to the identification of MSA and PSP. However, in a minority of patients no unequivocal diagnosis can be made using MRI findings alone. PMID- 10680807 TI - Mitochondrial DNA mutations in complex I and tRNA genes in Parkinson's disease. AB - OBJECTIVE: To identify mitochondrial DNA (mtDNA) mutations that predispose to PD. BACKGROUND: Mitochondrial complex I activity is deficient in PD. mtDNA mutations may account for the defect, but the specific mutations have not been identified. METHODS: Complete sequencing was performed of all mtDNA-encoded complex I and transfer RNA (tRNA) genes in 28 PD patients and 8 control subjects, as well as screening of up to 243 additional PD patients and up to 209 control subjects by restriction digests for selected mutations. RESULTS: In the PD patients, 15 complex I missense mutations and 9 tRNA mutations were identified. After screening additional subjects, rare PD patients were found to carry complex I mutations that altered highly conserved amino acids. However, no significant differences were found in the frequencies of any mutations in PD versus control groups. The authors were unable to confirm previously reported associations of mutations at nucleotide positions (np) 4336, 5460, and 15927/8 with PD. Complex I mutations previously linked to Leber's hereditary optic neuropathy, one of which has been linked to atypical parkinsonism, were not associated with PD. CONCLUSIONS: mtDNA mutations with a high mutational burden (present in a high percentage of mtDNA molecules in an individual) in complex I or tRNA genes do not play a major role in the risk of PD in most PD patients. Further investigations are necessary to determine if any of the rare mtDNA mutations identified in PD patients play a role in the pathogenesis of PD in those few cases. PMID- 10680808 TI - Objective changes in motor function during placebo treatment in PD. AB - OBJECTIVE: To examine the frequency, temporal development, and stability of objectively derived motor changes during placebo treatment in PD and to define the clinical domains and demographic groups most affected. BACKGROUND: Placebo effects are documented in neurology, but the timing and specific disabilities most susceptible to changes during placebo treatment in PD have not been examined. METHODS: The authors examined the placebo-treated group from a randomized, multicenter, placebo-controlled clinical trial of monotherapy ropinerole in PD patients without motor fluctuations. In 105 patients, they evaluated placebo-associated effects on the motor section of the Unified Parkinson's Disease Rating Scale (UPDRS), dividing the motor examination into four categories: tremor, bradykinesia, rigidity, and gait/balance/midline functions. The motor UPDRS and its subscales were compared over time (at baseline and at 4, 12, and 24 weeks) using Wilcoxon's signed rank test. They applied a rigorous definition of placebo-associated improvement as an improvement over baseline score in motor UPDRS of at least 50% or a change in at least two motor items at any one visit by > or =2 points. RESULTS: During the 6-month study, 16% of subjects improved on placebo treatment. The prevalence of response was steady (8 to 9%) at any one visit without a predominance of an early effect. No patient showed a placebo-associated improvement on all visits. All domains of parkinsonian disability were subject to placebo-associated improvement, with a trend toward more response in bradykinesia and rigidity than in tremor or gait/balance/midline function. Gender, age, disease duration, and baseline disability score did not influence the likelihood of improvement in association with placebo treatment. CONCLUSION: Based on a rigorous definition of placebo associated improvement, prominent improvements in objective measures of PD disability occur during clinical trials. Because placebo-associated improvements occur throughout a 6-month trial, placebo-controlled studies in PD should be at least 6 months to capture early as well as late improvements. PMID- 10680809 TI - Quantitative 1H MR spectroscopic imaging in early Rett syndrome. AB - OBJECTIVE: To determine cerebral regional concentrations of N-acetyl aspartate (NAA), total choline (Cho), and total creatine (Cr) in Rett syndrome (RS) using 1H magnetic resonance spectroscopic imaging (MRSI). BACKGROUND: The biochemical defect underlying RS is unknown. Because in vivo MRSI can detect important cerebral metabolites, MRSI has a potential to reveal impairment of regional cerebral metabolism in RS noninvasively. METHODS: High-resolution, multislice 1H MRSI was carried out in 17 girls with RS. The control group consisted of nine healthy children. RESULTS: In patients with RS, average Cho concentration was 12% higher (p < 0.005) and average NAA concentration 11% lower (p < 0.0001) compared with the control group. Regional metabolic differences included significantly lower NAA concentration in the frontal gray and white matter, insula, and hippocampus in RS; no difference in regional Cho and Cr concentrations were found. A 20 to 38% higher Cho:NAA ratio in frontal and parietal gray and white matter, insular gray matter, and hippocampus (p < 0.05) and a 14 to 47% lower NAA:Cr ratio in frontal cortical gray matter, parietal and temporal white matter, insula, and putamen (p < 0.05) were found in subjects with RS compared with controls. Patients with seizures had higher average concentrations of Cho, Cr, and NAA compared with those without seizures (8-19%, p < 0.05). CONCLUSION: Metabolic impairment in RS involves both gray and white matter and particularly involves frontal and parietal lobes and the insular cortex. Loss of NAA most likely reflects reduced neuronal and dendritic tree size; increased Cho concentration may result from gliosis. PMID- 10680810 TI - Vestibular hypersensitivity to sound (Tullio phenomenon): structural and functional assessment. AB - OBJECTIVES: To establish the role of high-resolution CT imaging and tests of vestibulocollic reflexes in diagnosing and understanding the pathogenesis of the Tullio phenomenon. BACKGROUND: The Tullio phenomenon is a syndrome in which acoustic stimulation produces symptoms and signs of vestibular activation. It has previously been associated with an abnormally low threshold for click-evoked vestibulocollic responses and also with dehiscence of the roof of the anterior (superior) semicircular canal on high-resolution CT scans of the temporal bones. METHODS: High-resolution CT scans of the temporal bones and vestibulocollic responses in sternocleidomastoid to both clicks and transmastoid galvanic stimulation (3 mA/2 msec) were studied in four patients with the Tullio phenomenon (one bilateral). RESULTS: Click-evoked thresholds were low for all affected ears (four at 65 dB nHL, one at 55 dB nHL) and normal (>70 dB nHL) for the three unaffected ears. In contrast, galvanic-evoked vestibulocollic responses were symmetric and of normal size in all patients. The bony roof of the anterior (superior) semicircular canal was thin, possibly absent, on CT of all affected ears and also in two out of three unaffected ears. CONCLUSIONS: The normal galvanic vestibulocollic responses indicate that sound sensitivity in patients with the Tullio phenomenon is likely to occur distal to the vestibular nerve, probably at the level of the receptors. Both click hypersensitivity and dehiscence of the anterior (superior) semicircular canal are associated with the Tullio phenomenon but as the CT scan abnormality can occur in clinically unaffected ears, click testing is important for specific diagnosis. Abnormal sound sensitivity, as demonstrated by click responses, confirms that the radiologic abnormality is function significant. PMID- 10680811 TI - Genetic analysis of vitamin D related genes in Canadian multiple sclerosis patients. Canadian Collaborative Study Group. AB - The objective of this study was to investigate genes involved in the metabolism and function of vitamin D as candidate genes for genetic susceptibility to MS. Restriction fragment length polymorphisms and highly polymorphic microsatellite markers within or very close to the 1,25(OH)2D3 receptor (VDR) [12q14], the vitamin D binding protein (DBP) [4q12], and the 25(OH)D2 1alpha-hydroxylase [12q13] loci were analyzed for linkage or association with MS. We found no evidence for linkage or association of these candidate genes with MS in the Canadian population. PMID- 10680812 TI - Anti-inflammatory drugs and Alzheimer-type pathology in aging. AB - Anti-inflammatory drugs have been suggested as a treatment for AD. The authors examined the AD-type pathology in postmortem brain tissue from elderly nondemented individuals who were chronically exposed to anti-inflammatory drugs. The results suggest that 1) these drugs do not affect the formation of either senile plaques or neurofibrillary tangles and 2) nonsteroidal anti-inflammatory drugs may be more effective than steroids in treating AD because of their ability to suppress the microglial activation associated with senile plaques. PMID- 10680813 TI - Cerebrospinal fluid pyruvate levels in Alzheimer's disease and vascular dementia. AB - Increased amounts of CSF pyruvate and lactate were found in patients with AD and patients with vascular dementia (VaD). In the AD group, CSF pyruvate values did not show any overlap with those obtained in controls; within the VaD group, the highest values were observed in possible VaD cases. A significant correlation between the severity of dementia and these biochemical parameters was also observed in both AD and possible VaD. The similarities of CSF pyruvate patterns observed in AD and possible VaD patients implicate a neurodegenerative component in this VaD subgroup. PMID- 10680814 TI - Amnesic syndrome with bilateral mesial temporal lobe involvement in Hashimoto's encephalopathy. AB - A 25-year-old woman presented with a subacute confusional state, headaches, unsteadiness, myoclonus, seizures, and an amnesic syndrome as a manifestation of Hashimoto's encephalopathy. Investigations showed biochemical hypothyroidism, raised thyroid microsomal antibodies, and weakly positive antineuronal antibodies. A T2-weighted MRI of the brain showed bilateral symmetric areas of increased signal in the mesial temporal lobes and hippocampi that had a low signal intensity on T1-weighted imaging. Despite clinical and radiologic improvement after steroid and thyroid hormone replacement therapy, a severe amnesic syndrome with associated localized MRI abnormalities persists. PMID- 10680815 TI - Frequency and duration of hospitalization of patients with AD based on Medicare data: CERAD XX. AB - Medicare records on 477 Consortium to Establish a Registry for Alzheimer's Disease patients with AD for 1991 through 1995 showed a hospitalization rate of 0.37/person-year with a length of stay of 3.7 days/ person-year (average of 10 days/hospitalization). Unmarried and less-educated patients with AD were admitted to the hospital more frequently, and, along with black patients, had a longer length of stay. Frequency and duration of hospitalization were greater in the patients with AD than in Medicare beneficiaries in general, but the rate of diagnostic/therapeutic procedures was lower. PMID- 10680816 TI - AIDS- and non-AIDS-related PML association with distinct p53 polymorphism. AB - A population-based analysis of progressive multifocal leukoencephalopathy (PML) showed PML frequencies of 5.1% among patients with AIDS and 0.07% among patients with hematologic malignancies, but similar clinical features of PML in both groups. Sequencing of the p53 gene, exon 4, showed heterozygosity (Arg-Pro) at codon 72 in five of six PML patients. These findings indicate that frequencies of non-AIDS- and AIDS-related PML differ markedly but p53 polymorphisms may influence the occurrence of PML in both groups. PMID- 10680817 TI - Positive CSF HSV PCR in patients with GBM: a note of caution. AB - Because two patients with temporal lobe glioblastomas had herpes simplex (HSV) DNA detected in CSF using PCR at the time of their presentation, we reviewed our laboratory's experience and performed PCR on a bank of 159 frozen CSF samples from patients with glioblastoma multiforme and other neurologic disorders. Based on the inability to detect HSV in any other tumor sample, we conclude that the positive HSV PCR in our two index patients most likely represented false-positive results. A diagnosis of HSE should not be made by PCR alone when the clinical presentation is atypical. PMID- 10680818 TI - Natural history of aortic arch atherosclerotic plaque. AB - To define the natural history of aortic arch plaque, we used B-mode ultrasonography to perform sequential study of the aortic arch. Eighty-nine patients were studied for up to 18 months. There was no change in 67% of total plaques; 77% of simple plaque (<4 mm) and 48% of complex plaque (> or =4 mm) did not progress. Atherosclerosis of the aortic arch can be sequentially studied with B-mode ultrasonography, and most of these lesions remain unchanged after up to 18 months of observation. PMID- 10680819 TI - Cerebral deep venous thrombosis presenting as acute micrographia and hypophonia. AB - Deep cerebral venous thrombosis is often a devastating condition associated with hemorrhagic infarction. We describe a patient who presented with acute micrographia and hypophonia as the sole manifestations of extensive deep venous sinus thrombosis. PMID- 10680820 TI - Acute hydrocephalus in nonketotic hyperglycinemia. AB - We present four patients with typical neonatal onset non-ketotic hyperglycinemia (NKH) who developed hydrocephalus requiring shunting in early infancy. Brain imaging revealed acute hydrocephalus, a megacisterna magna or posterior fossa cyst, pronounced atrophy of the white matter, and an extremely thin corpus callosum in all. The three older patients had profound developmental disabilities. This suggests that the development of hydrocephalus in NKH is an additional poor prognostic sign. PMID- 10680821 TI - Recombinant calcium channel is recognized by Lambert-Eaton myasthenic syndrome antibodies. AB - The authors studied sera from 36 patients with Lambert-Eaton myasthenic syndrome (LEMS) by immunoblots using the recombinant protein derived from the DNA sequence encoding for the domain III S5-S6 linker of the P/Q-type voltage-gated calcium channel al subunit. The results of 18 patients were positive for antibodies to this recombinant protein. The results of 2 of 10 patients with lung cancer without LEMS were also positive. PMID- 10680822 TI - Neurogenic stunned myocardium in Guillain-Barre syndrome. AB - Neurogenic stunned myocardium (NSM), a syndrome of reversible left ventricular dysfunction best described after subarachnoid hemorrhage, has not been associated with peripheral neuropathy. We describe a woman with Guillain-Barre syndrome in whom a syndrome compatible with NSM developed in the setting of a physiologically documented increase in sympathetic cardiovascular tone. This case supports the presumed unifying role of excessive sympathetic nervous system activation in the pathogenesis of NSM. PMID- 10680823 TI - "Tactile" sensory nerve potentials elicited by air-puff stimulation: a microneurographic study. AB - To investigate the sensory nerve responses to selective touch stimulation, sensory nerve action potentials after brief air-puffs were recorded with a microelectrode. In patients with peripheral neuropathy, those with impairment of tactile sensations had significantly smaller responses than did those without tactile impairment, suggesting receptor activation failure as well as nerve conduction failure. Brief air-puff stimulation, when combined with microneurography, could be used for evaluating the tactile receptor properties in humans. PMID- 10680824 TI - Cortical reorganization after acute unilateral hearing loss traced by fMRI. AB - Unilateral acoustic stimulation produces a functional MRI (fMRI)-blood oxygenation-level-dependent (BOLD) response mainly in the contralateral auditory cortex. In unilateral deaf patients, the BOLD response is bilateral. We studied a subject with sudden hearing loss after cochlear nerve resection before and repeatedly after surgery. During normal bilateral hearing, contralateral cortical BOLD responses were found. Progressing compensatory reorganization with bilateral representation of unilateral stimulation was detected over a period of approximately 1 year. PMID- 10680825 TI - The risk of abducens palsy after diagnostic lumbar puncture. PMID- 10680826 TI - Hashimoto encephalopathy: a brainstem vasculitis? PMID- 10680827 TI - Acute idiopathic dysautonomia: electrophysiology and response to intravenous immunoglobulin. PMID- 10680828 TI - Morvan's syndrome associated with voltage-gated K+ channel antibodies. PMID- 10680829 TI - POEMS syndrome with necrotizing vasculitis: a novel feature of vascular abnormalities. PMID- 10680830 TI - Arm levitation in progressive supranuclear palsy. PMID- 10680831 TI - Arm levitation in progressive supranuclear palsy. PMID- 10680832 TI - Ventricular asystole during vagus nerve stimulation for epilepsy in humans. PMID- 10680833 TI - Ventricular asystole during vagus nerve stimulation for epilepsy in humans. PMID- 10680834 TI - An active-control trial of lamotrigine monotherapy for partial seizures. PMID- 10680835 TI - The influence of smoking on the risk of Alzheimer's disease. PMID- 10680836 TI - Isolated Echinococcus granulosus hydatid cyst in the CNS with severe reaction to treatment. PMID- 10680837 TI - Effective gene therapy for pancreatic cancer by cytokines mediated by restricted replication-competent adenovirus. AB - Pancreatic cancer has a poor prognosis even when surgical treatment can be accomplished. Studies have demonstrated that pancreatic cancer is associated with various genetic abnormalities in oncogenes and tumor suppressor genes including p53. New therapeutic approaches for pancreatic cancer can be developed by targeting these genetic alterations. Adenovirus (Adv) lacking the 55-kDa E1B protein (E1B55K) replicates preferentially in p53-deficient cancer cells. We constructed E1B55K-deleted Adv (AxE1AdB), and studied its replication and cytopathic effect on pancreatic cancer cells. AxE1AdB replicated in and caused cell death of the p53-deficient pancreatic cancer cell lines tested (e.g., PANC 1, MIAPaCa-2, SU.86.86, BxPC-3, and PK-1). To enhance its therapeutic effect, we examined the combination of coinfecting this restricted replication-competent adenovirus (RRCA) with other Adv. Coinfection of E1-deficient Adv expressing the reporter lacZ gene (AxCAlacZ) together with AxE1AdB resulted in the replication of both viruses and a marked increase in reporter gene expression. PANC-1 cells coinfected with AxE1AdB and the Adv for human IL-2 (AxCAhIL2), produced 110 times more IL-2 than those infected with AxCAhIL2 alone. Similarly, coinfection of AxE1AdB and Adv for human IL-12 augmented the IL-12 production by 370-fold. Injecting AxE1AdB into the PANC-1 tumor of severe combined immunodeficient mice (SCID mice) resulted in marked reduction of the volume of the tumor. Moreover, injecting AxE1AdB with AxCAhIL2 into the PANC-1 tumor resulted in complete regression of the established tumors. These data suggest that RRCA, which augments the antitumor effect of a viral transgene (i.e., cytokines), may be a powerful tool for treating p53-deficient pancreatic cancer. PMID- 10680838 TI - Analysis of adenoviral transport mechanisms in the vessel wall and optimization of gene transfer using local delivery catheters. AB - Local delivery devices have been used for adenovirus-mediated gene transfer to the arterial wall for the potential treatment of vascular proliferative diseases. However, low levels of adenoviral gene expression in vascular smooth muscle cells may pose a serious limitation to the success of these procedures in the clinic. In this study, we examined the mechanisms controlling adenoviral transport to the vessel wall, using both hydrogel-coated and infusion-based local delivery catheters, with the goal of enhancing in vivo gene transfer under clinically relevant delivery conditions. The following delivery parameters were tested in vivo: applied transmural pressure, viral solution volume and concentration, and delivery time. We found that viral particles are transported into the vessel wall in a manner consistent with diffusion rather than pressure-driven convection. Consistent with diffusion, viral concentration was shown to be the key variable for viral transport in the vessel wall and thus gene expression in vascular smooth muscle cells. A transduction level of 17.8+/-3.2% was achieved by delivering a low volume of concentrated adenoviral beta-galactosidase solution through an infusion balloon catheter at low pressure without an adverse effect on medial cellularity. Under these conditions, effective gene transfer was accomplished within a clinically relevant time frame of 2 min, indicating that longer delivery times may not be necessary to achieve efficient gene transfer. PMID- 10680839 TI - The CXC chemokine, monokine induced by interferon-gamma, inhibits non-small cell lung carcinoma tumor growth and metastasis. AB - Angiogenesis is an absolute requirement for tumor growth beyond 2 mm3 in size. The balance in expression between opposing angiogenic and angiostatic factors controls the angiogenic process. The CXC chemokines are a group of chemotactic cytokines that possess disparate activity in the regulation of angiogenesis. Non small cell lung carcinoma (NSCLC) has an imbalance in expression of ELR+ (angiogenic) compared with ELR- (angiostatic) CXC chemokines that favors angiogenesis and progressive tumor growth. We found that the level of the ELR- CXC chemokine MIG (monokine induced by interferon gamma) in human specimens of NSCLC was not significantly different from that found in normal lung tissue. These results suggested that the increased expression of ELR+ CXC chemokines found in these tumor samples is not counterregulated by a concomitant increase in the expression of the angiostatic ELR-CXC chemokine MIG. This would result in an even more profound imbalance in the expression of regulatory factors of angiogenesis that would favor neovascularization. We hypothesized that MIG might be an endogenous inhibitor of NSCLC tumor growth in vivo and that reconstituion of MIG in the tumor microenvironment would result in the inhibition of tumor growth and metastasis. In support of this hypothesis, we demonstrate here that overexpression of the ELR-CXC chemokine MIG, by three different strategies including gene transfer, results in the inhibition of NSCLC tumor growth and metastasis via a decrease in tumor-derived vessel density. These findings support the importance of the ELR- CXC chemokine MIG in inhibiting NSCLC tumor growth by attenuation of tumor-derived angiogenesis. Furthermore, these findings demonstrate the potential of gene therapy as an alternative means to deliver and overexpress a potent angiostatic CXC chemokine. PMID- 10680840 TI - Catheter-mediated vascular endothelial growth factor gene transfer to human coronary arteries after angioplasty. AB - Blood vessels are among the easiest targets for gene therapy. However, no data are available about the safety and feasibility of intracoronary gene transfer in humans. We studied the safety and efficacy of catheter-mediated vascular endothelial growth factor (VEGF) plasmid/liposome (P/L) gene transfer in human coronary arteries after percutaneous translumenal coronary angioplasty (PTCA) in a randomized, double-blinded, placebo-controlled study. The optimized angioplasty/gene delivery method was previously shown to lead to detectable VEGF gene expression in human peripheral arteries as analyzed from amputated leg samples. Gene transfer to coronary arteries was done with a perfusion-infusion catheter, using 1000 microg of VEGF or beta-galactosidase plasmid complexed with 1000 microl of DOTMA:DOPE liposomes. Ten patients received VEGF P/L, three patients received beta-galactosidase P/L, and two patients received Ringer lactate. Gene transfer to coronary arteries was feasible and well tolerated. Except for a slight increase in serum C-reative protein in all study groups, no adverse effects or abnormalities in laboratory parameters were detected. No VEGF plasmid or recombinant VEGF protein was present in the systemic circulation after the gene transfer. In control angiography 6 months later, no differences were detected in the degree of coronary stenosis between treatment and control groups. We conclude that catheter-mediated intracoronary gene transfer performed after angioplasty is safe and well tolerated and potentially applicable for the prevention of restenosis and myocardial ischemia. PMID- 10680841 TI - Retroviral vector-mediated gene expression in human CD34+CD38- cells expanded in vitro: cis elements of FMEV are superior to those of Mo-MuLV. AB - A novel murine stromal cell line, HESS-M28, was established, which supports the expansion of human CD34+CD38- cells more than 300-fold in vitro in the presence of human IL-3 and SCF. These cells were used in an attempt to evaluate cis-acting elements of retroviral vectors in human primitive hematopoietic cells. Cord blood cells were cultured on top of the mixed cell layers of the stromal cell line, HESS-M28, and retroviral vector-producing cells. The FMEV-type vector SF/Lyt contained the spleen focus-forming virus U3 and the MESV primer-binding site (PBS), while MO3/Lyt contained the U3 region and PBS from Mo-MuLV. After transduction by the FMEV-type and Mo-MuLV-based vectors, expression of the marker gene murine CD8 (mCD8) was examined in CD34-, CD34+, and CD34+CD38- cells. In CD34+ and CD34+CD38- cells, expression of mCD8 was higher with the FMEV-type vector, SF/Lyt, compared with the cells transduced by the Mo-MuLV-based vector MO3/Lyt, although the expression was comparable in CD34- cells. Expression of marker genes was also confirmed in long-term culture-initiating cells (LTC-ICs) and SCID-repopulating cells (SRCs). PMID- 10680842 TI - Regressions of established breast carcinoma xenografts by carboxypeptidase G2 suicide gene therapy and the prodrug CMDA are due to a bystander effect. AB - The role of the bystander effect in the treatment of a human breast carcinoma xenograft was studied by suicide gene therapy with carboxypeptidase G2 (CPG2) and CMDA. Cells expressing enzymatically active surface-tethered bacterial CPG2 [stCPG2(Q)3] were mixed with control beta-galactosidase (beta-Gal)-expressing cells to give stCPG2(Q)3:beta-Gal ratios of, respectively: group 1, 0:100; group 2, 10:90; group 3, 50:50; and group 4, 100:0. Four days after injection of the cells into nude mice, the prodrug 4-[(2-chloroethyl)(2 mesyloxyethyl)amino]benzoyl-L-glutamic acid (CMDA) was administered. Tumor growth delay correlated well with the levels of stCPG2(Q)3 expression: group 1, 0 day delay; group 2, 10 days; group 3, 16 days; and group 4, 90 days. Similarly, the number of cures was strongly correlated to the levels of stCPG2(Q)3 activity: group 1, zero of six cured; group 2, one of six cured; group 3, three of six cured and group 4, four of six cured. There was a good correlation between CPG2 enzyme activity in the tumors and the number of cures. The majority of cells from groups 2 and 3 were apoptotic whereas those from group 1 were not, indicating a substantial bystander effect in the tumors. These results suggest that a bystander effect plays a major role in suicide gene therapy regimens with stCPG2(Q)3 and CMDA. PMID- 10680843 TI - Targeting human T cells by retroviral vectors displaying antibody domains selected from a phage display library. AB - To generate T cell-specific retroviral vectors an scFv phage display library derived from immunized mice was selected for binding to the human T cell line Molt-4/8. The scFv cDNAs recovered from the selected phages were transiently expressed as an N-terminal fusion of the spleen necrosis virus (SNV) transmembrane protein (TM) subunit of the viral envelope protein (Env) in the cell line DSH-cxl, which packages the beta-galactosidase gene into SNV particles. Screening of supernatants from about 150 transfections resulted in the identification of 5 scFvs that mediated efficient transduction of Molt-4/8 cells. Using stable packaging cell lines vector preparations with titers greater than 10(4) EFU/ml on human T cells were obtained. The scFv 7A5 in particular was able to mediate selective transduction of human T cells with high efficiency. Titers of up to 106 EFU/ml were reached on Molt-4/8, Jurkat, and A301 cells, while titers on HeLa cells, TE671 cells, 293T cells, and HT1080 cells were below 102 EFU/ml. Transduction of stimulated primary human peripheral blood cells, which consisted mainly of T cells, was about fivefold more efficient than transduction of B cells. Western blot analysis of supernatant from the 7A5 packaging cells demonstrated incorporation of 7A5-TM into vector particles and indicated proteolytic processing of the coexpressed unmodified TM during particle formation. Binding of bacterially expressed 7A5-scFv to a panel of cell lines correlated well with the transduction results. These data provide the first proof of concept that a general approach can be taken to obtain scFvs able to mediate selective gene transfer into target cells. PMID- 10680844 TI - Macrophage colony-stimulating factor can modulate immune responses and attract dendritic cells in vivo. AB - Studies have indicated that professional APCs in the periphery, such as dendritic cells and macrophages, play an important role in initiating DNA vaccine-specific immune responses. To engineer the immune response induced by DNA vaccines in vivo we investigated the modulatory effects of codelivering growth factor genes for the hematopoietic APCs along with DNA vaccines. Specifically, we examined the effects on the antigen-specific immune responses following the codelivery of the gene expression cassettes for M-CSF, G-CSF, and GM-CSF along with HIV-1 DNA immunogen constructs. We observed that coimmunization with GM-CSF increased the antibody response and resulted in a significant enhancement of lymphoproliferative response. Furthermore, among all coinjection combinations, we found that M-CSF coinjections resulted in a high level of CTL enhancement. This enhancement of CTL responses observed from the coinjection with M-CSF was CD8+ T cell dependent and was associated with the presence of CD11c+ cells at the site of injection and with the antigen-specific induction of the beta-chemokine MIP 1beta, suggesting a role for this chemokine in CTL induction. These results suggest that hematopoietic growth factors should be further studied as potential adjuvants for in vivo modulators of immune responses. PMID- 10680845 TI - Lentiviral and murine retroviral transduction of T cells for expression of human CD40 ligand. AB - Efficient transduction of primary human T cells is an important goal toward treating a number of genetic defects. Patient T cells could be harvested by leukapheresis, transduced, and returned to the donor. A wide range of secreted or cell surface therapeutic proteins may be delivered in this way. The ability to produce antibodies is the consequence of interactions between T cells and B cells and lack of expression of CD40 ligand (CD40L) on T cells causes X-linked hyper IgM syndrome (XHIM). We are investigating delivery of a normal CD40 ligand to treat this disorder. We tested promoters driving the expression of either reporter genes such as enhanced green fluorescent protein (eGFP) or human CDC40L. Using murine retroviruses, the best able to drive gene expression in T cells was the cytomegalovirus (CMV) promoter enhancer element; however, transduction efficiency was low. To achieve efficient, high-level gene expression we tested lentiviral gene delivery vectors. At a low multiplicity of infection (MOI) (0.5 2) a large fraction of target cells was transduced by lentiviral vectors (40 93%), and the strength of gene expression was high, as determined by flow cytometric analysis. We monitored the expression of eGFP or human CD40L on T cell lines and untransformed primary human T cells from normal and CD40L-deficient patients. We achieved efficient gene expression without an extended exposure to virus, and without the need for selection. These results are encouraging for efficient lentivirus-mediated transduction of refractory human cells to achieve therapeutic gene delivery. PMID- 10680846 TI - DeltahGHR, a novel biosafe cell surface-labeling molecule for analysis and selection of genetically transduced human cells. AB - We describe a new selectable marker for retroviral transduction and selection of human and murine cells. The molecule expressed on the cell surface of the transduced population is a truncated version of human growth hormone receptor (deltahGHR), capable of ligand (hGH) binding, but devoid of the domains involved in signal triggering. We demonstrate that the engineered molecule is stably expressed in the target cells as an inert protein unable to trigger proliferation or to rescue the cells from apoptosis after ligand binding. This new marker will probably have a wide application spectrum, since hGHR in the human adult is highly expressed only in liver cells, and lower levels have been reported in certain lymphocyte cell populations. The deltahGHR label has high biosafety potential, as it belongs to a well-characterized hormonal system that is nonessential in adults, and there is extensive clinical experience with hGH administration in humans. This record allows us to hypothesize the lack of relevant clinical consequences resulting from massive transgene expression caused by successful replacement of a large tissue with genetically transduced cells. We take advantage of the differential binding properties of several monoclonal antibodies (MAbs) in describing a cell rescue method in which the antibody used to select deltahGHR-transduced cells is eluted by competition with hGH or, alternatively biotinylated hGH is used to capture tagged cells. In the latter system, the final purified population would be recovered free of attached antibodies in hGH (a substance approved for human use)-containing medium, providing additional biosafety relative to currently existing methods that rely on the use of murine MAb to rescue genetically labeled cells. PMID- 10680847 TI - Anti-human immunodeficiency virus type 1 gene therapy in human central nervous system-based cells: an initial approach against a potential viral reservoir. AB - Studies have demonstrated that human immunodeficiency virus type 1 (HIV-1) infection of central nervous system (CNS)-based cells in vivo results in a series of devastating clinical conditions collectively termed acquired immune deficiency syndrome (AIDS) dementia complex (ADC). Gene therapy for these neurovirological disorders necessitates utilization of a vector system that can mediate in vivo delivery and long-term expression of an antiretroviral transgene in nondividing/postmitotic CNS cellular elements. The present studies focus on the transfer of an anti-HIV-1 gene to primary isolated CNS microvascular endothelial cells (MVECs) and neuronal-based cells, for its effects in protecting these cells from HIV-1 infection. By using an HIV-1-based vector system, it was possible to efficiently transduce and maintain expression of a marker transgene, beta galactosidase (beta-Gal), in human CNS MVECs, human fetal astrocytes, plus immature and mature (differentiated) NT2 cells. Significant transduction of the marker gene, beta-Gal, in CNS-based cells prompted the utilization of this system with an anti-HIV-1 gene therapeutic construct, RevM10, a trans-dominant negative mutant Rev protein. Initially, it was not possible to generate any HIV-1 vector particles with the RevM10 gene in the transducing construct, because of inhibitory effects on the HIV-1 vector by this gene product. However, the vector could be partially rescued by adding an additional construct that supplied wild type rev, in trans, during a multiple construct transfection in the packaging 293T cells. Thus, it was possible to significantly improve the titer of RevM10 expressing viral particles generated from these cells. Moreover, this RevM10 vector transduced the neuronal precursor cell line NT2, retinoic acid differentiated human neurons (hNT) from the precursor cells, and primary isolated human brain MVECs with high efficiency. RevM10 generated from the HIV-1-based vector system potently inhibited replication of diverse HIV-1 strains in human CNS MVECs and neuronal cells. The data generated from these studies represent an initial approach for future development of anti-HIV-1 gene therapy in the CNS. PMID- 10680848 TI - Coarse spray delivery to a localized region of the pulmonary airways for gene therapy. AB - Targeting adenoviral vectors for cystic fibrosis gene therapy to the human airways with minimal exposure to alveoli would avoid adverse reactions and maximize response. At present, to deliver gene therapy vectors, large volumes of fluid are instilled or nebulized as aerosols. Either approach would likely cause alveolar exposure and increases the potential for side effects. We describe a coarse spray delivery device that precisely and reproducibly delivers the viral vector to the human airways to treat a small region of the airways for clinical trials. An endoscopic washing pipe (Olympus) that can be inserted into the channel of a bronchoscope was used. To minimize the escape of the therapeutic material downstream from the site of administration, we restricted the volume delivered to <150 microl (to prevent bulk flow), and used large droplets. Their high velocity further enhanced the probability of impaction in the vicinity of the nozzle. A pneumatic dosing system (Kahnetics) was used to reproducibly deliver the spray. The droplet size distribution was determined by laser diffraction and confirmed by cascade impaction: 190-microm volume median diameter with 1% mass <10 microm. The localization of the spray was studied in hollow cast models of human airways. 99mTc-sulfur colloid was used as a radiolabeled marker for these studies. Localization of the deposited spray was determined by scintigraphy and by measuring the radioactivity exiting the terminal airways. In the lung casts the spray was localized to one or two generations over an approximately 2-cm2 area. We conclude that delivery of large droplet sprays limits exposure to a few generations and may be useful in topical gene delivery clinical trials. PMID- 10680849 TI - Primary care options to prevent mental illness. AB - The common mental disorders, mainly anxiety and depression, constitute a major public health problem, incurring considerable costs in terms of use of health services and time lost from work. Risk factors include low socioeconomic status, poverty and poor housing, as well as stressful life events and difficulties such as demanding child care, separation or divorce, bereavement, loss of employment and caring for a dependant relative. Population approaches are probably necessary to reduce significantly the burden of such mental health problems, but health care measures are far from negligible. Primary care professionals have regular opportunities to identify people at risk of mental health problems and refer them to welfare and social support services (primary prevention). A number of interventions among high-risk groups have been shown to be effective, including problem-solving training and cognitive-behavioural approaches. The most important tasks in primary care are to identify people with depression, alcohol and drug misuse and eating disorders as early as possible in the course of their illness and to institute effective treatment (secondary prevention). Primary care teams should also join in shared care arrangements for patients with chronic disabling mental illnesses, in order to prevent recurrences and relapses (tertiary prevention). PMID- 10680851 TI - Development of gender differences in depression: description and possible explanations. AB - This article reviews the description and possible explanations for the development of gender differences in depression in children and adolescents. The emerging gender difference (more girls depressed than boys) in depressed mood and depressive disorders appears after the age of 13 years or midpuberty. Currently, little evidence supports that biological factors are an explanation. Genetic factors are associated more strongly with depression among pubertal girls than boys. Regarding cognitive factors, ruminative response style, but not dysfunctional attitudes or attributional style, has been supported to be a possible explanation. Studies on childhood adversities and gender role have provided evidence explaining why more girls are depressed than boys. Girls are more likely to experience negative events in the family than boys, and these adversities are in turn associated with elevated depression. Girls identify more strongly with a feminine stereotype of needing to appear thin and consequently become more dissatisfied with their body shape and physical appearance, which in turn is associated with increased depression. PMID- 10680850 TI - Euthanasia and physician-assisted suicide. AB - In the Netherlands there are about 9700 explicit requests for euthanasia or physician-assisted suicide (EAS) each year, of which approximately 3600 are granted. Other countries have criticized the Dutch policy concerning EAS. It has been suggested that palliative care in the Netherlands is not adequate and that euthanasia is often requested by patients with depression. In addition, this criticism is partly based on the firm stance that 'human life has an absolute value and a human being has under no circumstances the right of self determination over his or her own life'. Many aspects of EAS are currently the focus of attention in the literature. In this review the following aspects of EAS are discussed: ethics, judicial questions, the relationship between depression and euthanasia, and the impact of EAS on members of the family. Also, the current situation concerning EAS in the Netherlands is summarized and described. Despite the fact that EAS have been widely discussed in the literature, the association between depression and the number of requests for EAS remains to be discovered. It is also not yet known what the effects of EAS are on members of the family, and whether unnatural death causes a higher incidence of complicated grief. PMID- 10680852 TI - Memory retrieval under the control of the prefrontal cortex. AB - Memory retrieval is a process wherein a distributed neural network reactivates the brain's representation of past experiences. Sensory long-term memory is represented among a population of neurones in the modality-specific posterior association cortex. The coded representation of memory can be retrieved by interactions of hierarchically different cortical areas along bottom-up and top down anatomical connections. We examined the function of the prefrontal cortex in memory retrieval by two different approaches. Firstly, a meta-analysis of brain imaging studies revealed that the prefrontal cortex is reliably activated by memory retrieval in humans. Secondly, in order to determine the causal relationship between the prefrontal activations and memory retrieval, we designed a new experimental paradigm using posterior-split-brain monkeys. Following section of the splenium of the corpus callosum and the anterior commissure, visual stimulus-stimulus association learning within one hemisphere did not transfer to the other. Nevertheless, when a visual cue was presented to one hemisphere, the prefrontal cortex could instruct the contralateral hemisphere to retrieve the correct stimulus specified by the cue. These findings suggest that the prefrontal cortex can regulate the recall of long-term memory in the absence of bottom-up sensory inputs. PMID- 10680853 TI - Medical treatment of erectile dysfunction. AB - Erectile dysfunction (ED) is defined as the consistent inability to obtain or maintain an erection for satisfactory sexual relations. Data from the Massachusetts Male Aging Study have indicated that the prevalence of erectile dysfunction of any degree is 39% in 40-year old men, and 67% in those aged 70 years. Effective therapy has been available for some time, but it has commonly involved surgery, external devices or penile self-injection. For many men, these represent unacceptable barriers to seeking therapy. Recently, however, an effective oral medication has become available. This article reviews the physiology and pharmacology of ED. The literature currently available on the effectiveness and safety of various drugs used for ED is summarized, with particular attention to newly available oral agents. Guidelines for work-up and drug treatment of patients with ED are given. Detailed history and physical examination are crucial to the safe and effective treatment of men with erectile impotence. An extensive review of the literature shows that based on safety, effectiveness and ease of use, oral sildenafil citrate is an excellent choice for first-line therapy. Patients who use organic nitrates of any kind in any capacity should not be offered sildenafil. Based solely on effectiveness intracavernosal injection therapy remains the golden standard and should also be offered as an option for first-line therapy for the appropriate patients. Many alternatives are available for men who cannot use sildenafil or injection therapy. A thorough knowledge of existing medications is essential for proper treatment of ED. PMID- 10680854 TI - New diagnostic findings in coeliac disease. AB - Coeliac disease, a life-long gluten-sensitive disorder, characterized by malabsorption, villous atrophy and crypt hyperplasia, is well recognized. However, the disease is evidently underdiagnosed, and the classic forms constitute only the tip of the 'coeliac iceberg'. Patients with coeliac disease can have subtle symptoms, if any. Diagnostic difficulties may further emerge when minor mucosal changes are found. In coeliac screening and case-finding a novel test, the antitissue transglutaminase test, has proven promising with a sensitivity and specificity of over 95 %. Genetic and immunohistological research has taken a great leap forward. Coeliac disease is strongly associated with HLA DQ2, coded by the DQA1*0501 and DQB1*02 alleles, or the DQ8 (DQA1*03, DQB1*0302 alleles). The disease is rare in patients who do not share these alleles, a circumstance which can be utilized in diagnostics. An increase in small bowel intraepithelial lymphocytes especially gammadelta+ T-cell receptor-bearing cells is typical, albeit not pathognomonic, for coeliac disease. Combining new symptoms, humoral immunity, genetics and immunohistological staining can today offer a greater diagnostic scope for coeliac disease, especially in cases where clinical presentation and small bowel biopsy findings remain doubtful. PMID- 10680855 TI - Neurological channelopathies: diagnosis and therapy in the new millennium. AB - Rapid progress in the complementary fields of molecular genetics and cellular electrophysiology has led to a better understanding of many disorders which are caused by ion channel dysfunction. These channelopathies may manifest in a multitude of ways depending on the tissue specificity of the channel that is affected. Several important general medical conditions are now known to be channelopathies but the neurological members of this family are amongst the best characterized. Over recent years, ion channel dysfunction in skeletal muscle in particular has emerged as a paradigm for understanding neurological ion channel disorders. This review concentrates mainly on the diseases caused by dysfunction of the voltage-gated ion channels. We initially focus on the skeletal muscle channelopathies (the periodic paralyses, malignant hyperthermia, paramyotonia congenita and myotonia congenita). The central nervous system channelopathies are then explored, with particular reference to the advances which have implications for understanding the mechanisms of common neurological disorders such as epilepsy and migraine. Looking towards the new millennium, DNA-based diagnosis will become a realistic proposition for most neurological channelopathies. Furthermore, it seems likely that new therapies will be designed based on genotype and mode of ion channel dysfunction. PMID- 10680856 TI - In situ use of suicide genes for therapy of brain tumours. AB - Suicide gene therapy represents a new therapeutic approach to the treatment of patients with otherwise incurable malignant brain tumours. This strategy involves the introduction of a gene that renders the tumour cell susceptible to an otherwise nontoxic prodrug. The most often used genetic prodrug activation system is the herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) paradigm. An important aspect of this system is the 'bystander effect', the extension of cytotoxic effects to untransduced cells. For gene delivery, retroviral, adenoviral vectors and HSV-1 mutants have been used. Clinical studies have revealed that the HSV-tk/GCV approach is safe, but also that responses are observed only in very small brain tumours, indicating insufficient vector distribution and very low transduction efficiency with replication-deficient vector systems. To improve treatment efficacy, the use of replication-competent oncolytic vectors in combination with new or improved prodrug-suicide gene systems as a part of a multimodal approach is warranted. In the context of replication-competent vectors, suicide genes might also be used as fail-safe genes in the case of runaway infection. PMID- 10680857 TI - PET in drug discovery and development: an introduction. PMID- 10680858 TI - Drug development, radiolabelled drugs and PET. AB - Positron emission tomography (PET) provides noninvasive in vivo quantitative pharmacokinetic and pharmacodynamic information on novel and established drugs. Because only very low amounts of the (potential) drug have to be administered, far below toxicity levels, human studies can be carried out even before the drug is entered in phase I studies. Such studies can provide cost-effective predictive toxicology data and information on the metabolism and mode of action of drugs. PET is also very useful in the study of the metabolic consequences of gene expression or gene defects. In the last decade, several models using genetically engineered small animals have been developed. The study of these animals with high-resolution small animal PET cameras provides new opportunities in drug development. Especially valuable is the contribution of PET in bridging the gap between molecular biology, basic pathology and the design of a new generation of drugs. PMID- 10680859 TI - Ligand-receptor interactions as studied by PET: implications for drug development. AB - Positron emission tomography (PET) is a quantitative imaging method that can be used to characterize binding properties of specific target molecules such as various receptors, transporter molecules and enzymes in vivo. Although already applied successfully, one of the greatest challenges for the technique is to understand better the in vivo complexities of ligand-receptor (target) interaction. The PET technique can be used efficiently in animal studies but, most importantly, also in human studies. PET imaging of patients and healthy volunteers can generate information on human pathophysiology at a molecular level currently unobtainable with other methods. Modern imaging techniques are increasingly applied to drug discovery and development. There are many ways of utilizing PET in pharmacodynamic studies, one interesting approach being the indirect exploration of synaptic neurotransmission with receptor ligands. The receptor occupancy-type studies with PET are rapidly becoming a state-of-the-art method for verifying the mechanism of action of a given drug in man and especially for facilitating the dose-finding procedures in early drug development. Thus far, PET has been mainly applied to pharmacodynamic studies in the central nervous system but will be used also in other areas of drug development such as cardiovascular diseases and oncology. PMID- 10680860 TI - Drug development for neurodegenerative diseases: role of PET. AB - There are few relevant animal models for neurodegenerative diseases to be used for human drug development. Most current drugs for neurodegenerative diseases act through different neurotransmitter systems. Positron emission tomography (PET) is a unique tool in the study of neurodegenerative diseases as it enables quantitative measurements of oxygen consumption, blood flow, energy metabolism and functioning of various neurotransmitter systems. There are several possibilities in the use of PET in drug development. It is possible to radiolabel the drug itself or to study the effect of an unlabelled drug on blood flow, energy metabolism or function of neurotransmitter systems. All these approaches have been used in drug development for neurodegenerative diseases. However, in spite of the important role of PET in pathophysiological studies of neurodegenerative diseases, thus far the versatile possibilities of PET in drug development for neurodegenerative diseases have not been fully exploited. PMID- 10680861 TI - PET as a cardiovascular and metabolic research tool. AB - Positron emission tomography (PET) represents the most advanced scintigraphic imaging technology. It can be employed for cardiovascular research as well as for clinical applications in patients with various cardiovascular diseases. PET allows the noninvasive functional assessment of myocardial perfusion, substrate metabolism and cardiac innervation as well as the study of skeletal muscle metabolism and perfusion in vivo. The large number of existing tracers and the flexibility of the PET technique that allows it to be combined with many other methods, such as the insulin clamp technique, increase its potential as a research tool. In the detection of myocardial viability PET is regarded as the golden standard, and it is the only method available for the quantitative assessment of myocardial blood flow. PMID- 10680862 TI - Temporal lobe injury in temporal bone fractures. AB - OBJECTIVE: To determine the incidence of intracranial injury, specifically in the temporal lobe, in patients with longitudinal fractures of the temporal bone. DESIGN: Prospective inception cohort. SETTING: University of Maryland Division of Otolaryngology-Head and Neck Surgery and the Maryland Shock Trauma Center, Baltimore. PATIENTS: Twenty-seven consecutive patients with unilateral or bilateral temporal bone fractures. MAIN OUTCOME MEASURES: Evaluation of temporal bone and intracranial trauma using computed tomography (CT) and magnetic resonance imaging (MRI). RESULTS: Of the 27 patients enrolled in the study, 12 had the complete battery of MRI, CT, and physical and audiological examinations. In all 12 patients, MRI demonstrated adjacent middle cranial fossa meningeal enhancement. Results of non-contrast-enhanced CT and MRI demonstrated ipsilateral temporal lobe contusions in 6 of the 13 fractures for an overall incidence of 46%. In addition, MRI demonstrated 4 cerebral contusions not seen in the results of non-contrast-enhanced CT. CONCLUSIONS: While high-resolution CT remains the criterion standard for evaluation of temporal bone fractures, MRI revealed a higher incidence of related temporal lobe injuries. Magnetic resonance imaging data may be valuable in preoperative evaluation of patients who require surgical intervention through a middle cranial fossa approach to document pre-existing injury and potential morbidity before retraction of the middle cranial fossa dura mater and temporal lobe. PMID- 10680863 TI - Dehiscence or thinning of bone overlying the superior semicircular canal in a temporal bone survey. AB - OBJECTIVE: To determine the incidence and etiology of dehiscences of bone overlying the superior semicircular canal in a temporal bone archive. DESIGN: A microscopic study was performed of 1000 temporal bones from 596 adults in a university hospital registry. Specimens were sectioned vertically in the plane of the superior semicircular canal. Measurements of minimum bone thickness over the superior canal were made in a subset of 108 randomly chosen specimens. All bones were examined for thinning or dehiscence relative to these norms. Clinical histories, when available, were reviewed. RESULTS: Complete dehiscence of the superior canal was identified in 5 specimens (0.5%), at the middle fossa floor (n = 1) and where the superior petrosal sinus was in contact with the canal (n = 4). In 14 other specimens (1.4%), the bone at the middle fossa floor (n = 8) or superior petrosal sinus (n = 6) was no thicker than 0.1 mm, significantly less than values measured in the control specimens (P<.001). Abnormalities were typically bilateral. Specimens from infants demonstrated uniformly thin bone over the superior canal in the middle fossa at birth, with gradual thickening until 3 years of age. CONCLUSIONS: Dehiscence of bone overlying the superior canal occurred in approximately 0.5% of temporal bone specimens (0.7% of individuals). In an additional 1.4% of specimens (1.3% of individuals), the bone was markedly thin (< or =0.1 mm), such that it might appear dehiscent even on ultra-high resolution computed tomography of the temporal bone. Sites affected were in the middle fossa floor or a deep groove for the superior petrosal sinus, often bilaterally. These abnormalities may arise from failure of postnatal bone development. Thin areas of bone over the superior canal may be predisposed to disruption by trauma. PMID- 10680864 TI - Neuroprotection due to irrigation during bipolar cautery. AB - OBJECTIVE: To test whether irrigation during bipolar cautery confers thermoprotection from neuronal injury. DESIGN: A rat animal model (15 rats for each treatment group) was used to test the thermoprotective effects of irrigation during bipolar cautery. In this model, the sciatic nerve was exposed, and a 1 second stimulus was applied using bipolar cautery forceps at 40 or 20 W placed directly on the nerve in the presence or absence of simultaneous irrigation. The effects of cautery were determined on the basis of clinical gait analysis by means of the Sciatic Functional Index, temperature response, and neuropathological findings. RESULTS: The degree of paresis was reduced with irrigation. Neuropathological examination of the sciatic nerve after cautery showed significant axonal loss (more small than large fibers) with concomitant demyelination, which was partially inhibited by irrigation (chi2; P = .04). The mechanism of thermoprotection by irrigation was not the result of a reduction in the temperature spike that followed cautery, but resulted from a reduced temperature response during the 15 seconds that followed 40- or 20-W stimulation with bipolar cautery. CONCLUSIONS: Simultaneous irrigation and bipolar cautery enhance temperature recovery to basal levels and protect the peripheral nerve from the effects of cautery. PMID- 10680865 TI - Paranasal sinus development and choanal atresia. AB - BACKGROUND: Although the determinates of paranasal sinus development and sinusitis are not well defined, a candidate factor is blockage of the choana. HYPOTHESIS: Maxillary sinuses ipsilateral to unilateral choanal atresia are comparatively small and have more evidence of sinusitis than do the contralateral sinuses. DESIGN: Retrospective. SETTING: Children's hospitals. PATIENTS: Sixteen nonsyndomic children with isolated unilateral congenital choanal atresia. MAIN OUTCOME MEASURES: Determination of maxillary sinus volumes and mucoperiosteal thickening on preoperative computed tomograms. RESULTS: Maxillary sinuses ipsilateral to unilateral choanal atresia have slightly larger volumes than, and mucoperiosteal thickening that is similar to, the contralateral sinuses. CONCLUSION: These data suggest that maxillary sinus development and sinusitis are independent of posterior nasal ventilation and drainage. PMID- 10680866 TI - Adenotonsillar enlargement in pediatric patients following solid organ transplantation. AB - OBJECTIVE: To evaluate the management of adenotonsillar hypertrophy in pediatric patients after transplantation. DESIGN: A retrospective medical record review after transplantation of all pediatric patients undergoing adenotonsillectomy at the University of California, Los Angeles, Medical Center during a 14-month period. SETTING: A tertiary care center. PATIENTS: There were 16 patients in our review, 11 boys and 5 girls. Nine patients had undergone liver transplantation, and 7 had undergone kidney transplantation. INTERVENTION: Fourteen patients underwent adenotonsillectomy, and 2 underwent adenoidectomy alone. Indications for surgical intervention included progressive symptoms of upper airway obstruction, recurrent tonsillitis, and/or evidence of notable adenotonsillar enlargement on physical examination. RESULTS: The mean +/- SD age at the time of transplantation was 3 years 1 month +/- 3 years 5 months. The mean +/- SD duration from allograft transplantation to adenotonsillectomy was 5 years 1 month +/- 2 years 4 months. Histopathologic examination revealed that 1 kidney transplant recipient had posttransplantation lymphoproliferative disorder. Eleven patients were found to have Epstein-Barr virus-related lymphoid hyperplasia. All patients experienced clinical resolution of their symptoms after surgery. CONCLUSIONS: Posttransplantation lymphoproliferative disorder is a condition associated with the Epstein-Barr virus infection in the setting of immunosuppression. Early presentation of posttransplantation lymphoproliferative disorder in children may be manifested by adenotonsillar enlargement. In addition to the role in relieving upper airway obstruction and decreasing upper respiratory tract infection, adenotonsillectomy may be critical in the prompt evaluation and treatment of posttransplantation lymphoproliferative disorder. PMID- 10680867 TI - Treatment of chronic suppurative otitis media with topical tobramycin and dexamethasone. AB - OBJECTIVES: To investigate the safety and efficacy of a topical combination of tobramycin and dexamethasone in a primate model of chronic suppurative otitis media (CSOM) and to explore the contribution of the added topical steroid for the treatment of CSOM. DESIGN: Blinded, randomized, placebo-controlled trial. SUBJECTS: Sixty juvenile cynomolgus monkeys randomized into the following 6 treatment groups of 10 monkeys each: 0.3% tobramycin (group 1), combined 0.3% tobramycin-0.1% dexamethasone (group 2), combined 1.0% tobramycin-0.33% dexamethasone (group 3), 0.1% dexamethasone (group 4), vehicle (group 5), and phosphate-buffered saline solution (group 6). INTERVENTIONS: Chronic suppurative otitis media was established by inoculating the right ear with Pseudomonas aeruginosa. After 4 weeks of drainage, animals were treated according to the group assignment with 3 drops twice daily for 7 weeks. Hearing thresholds were monitored with repeated auditory brainstem response testing (ABR), and clinical response was monitored with repeated otoscopic examinations and cultures throughout the study. Cytocochleograms were evaluated for quantification of outer hair cell loss. RESULTS: Rapid resolution of otorrhea and eradication of P aeruginosa occurred in all groups receiving tobramycin. The inclusion of dexamethasone accelerated the resolution of otorrhea and negative yields of cultures compared with tobramycin alone. Otorrhea and positive culture findings persisted in the groups not treated with topical antibiotic. Results of ABRs at 4 and 8 weeks and cytocochleograms for outer cell hair loss were not affected by drug administration. Perilymph samples collected at the end of the study showed no detectable tobramycin. CONCLUSIONS: Combined tobramycin-dexamethasone ear drops were safe and effective in the monkey CSOM model. Dexamethasone enhanced the efficacy of tobramycin. PMID- 10680868 TI - Aerobic and anaerobic bacteriology of concurrent chronic otitis media with effusion and chronic sinusitis in children. AB - OBJECTIVE: To correlate the aerobic and anaerobic microbiologic findings of concurrent chronic otitis media with effusion and chronic maxillary sinusitis. METHODS: Cultures were obtained from 32 children with concurrent chronic otitis media with effusion and maxillary sinusitis who underwent tympanostomy tube placement. RESULTS: A total of 42 isolates, 24 aerobic and 18 anaerobic, were recovered from 30 patients; 27 were isolated from both sites, 4 from the ear only, and 11 from the sinus only. The most common isolates were Haemophilus influenzae (9 isolates), Streptococcus pneumoniae (n = 7), Prevotella species (n = 8), and Peptostreptococcus species (n = 6). Microbiological concordance between the ear and sinus was found in 22 (69%) of culture-positive patients. CONCLUSION: The concordance in recovery of organisms in more than two thirds of the patients illustrates the common bacterial etiology between chronic otitis media with effusion and chronic sinusitis in children. PMID- 10680869 TI - Pediatric parotid masses. AB - OBJECTIVE: To evaluate the incidence, types, and treatment outcomes of pediatric parotid lesions. DESIGN: Retrospective case review, histological tissue review, and literature review. SETTING: Tertiary care center. PATIENTS: All patients aged 18 years and younger with parotid masses evaluated and treated at the Mayo Clinic, Rochester, Minn, from January 1, 1970, to December 31, 1997. RESULTS: Parotid masses were identified in 118 children (60 boys and 58 girls). At diagnosis, the ages of patients were from birth through 18 years, and 72 (61.0%) were aged 10 years and older. An asymptomatic mass was the most common presentation. Forty-three patients (36.4%) had infectious or inflammatory lesions, 56 (47.5%) had benign lesions, and 19 (16.1%) had malignant lesions. The most common benign lesions were pleomorphic adenoma (22.9%) and hemangioma (10.2%). The most common malignant lesions were mucoepidermoid carcinoma (6.8%) and acinic cell carcinoma (3.4%). The most common treatment was total parotidectomy (40.7%). Surgical complications included temporary facial nerve weakness in 22 (18.6%) patients, permanent facial weakness in 11 (9.3%), and permanent paralysis in 2 (1.7%). Pleomorphic adenoma recurred in 4 (14.8%) of 28 patients and mucoepidermoid carcinoma in 3 (37.5%) of 8 patients. One patient with adenoid cystic carcinoma died of the tumor. CONCLUSIONS: Although pediatric parotid masses are unusual, they can represent a variety of pathological diagnoses, including malignancy. We advocate prompt evaluation and treatment of these masses, and suggest guidelines for their management, based on diagnosis. PMID- 10680870 TI - Oropharyngotonsillitis associated with nonprimary Epstein-Barr virus infection. AB - OBJECTIVE: To identify distinct clinical features of pharyngotonsillitis or oropharyngitis associated with Epstein-Barr virus (EBV) infection from herpes simplex virus infection. DESIGN: Clinical studies by case exploration. SETTING: Institutional practice at a university hospital. PATIENTS: Thirty-three patients with pharyngotonsillitis and 4 patients with oropharyngitis of nonbacterial infection underwent biopsy of pharyngotonsillar lesions. MAIN OUTCOME MEASURE: The specimens were examined by histopathology, immunohistochemistry, in situ hybridization, and polymerase chain reaction. In addition to serological testing and routine laboratory data, photographic oropharyngeal findings were collected for clinical evaluation. RESULTS: In situ hybridization to detect EBV-encoded small nuclear RNA-1 and -2 disclosed 8 cases of pharyngotonsillitis and 4 cases of oropharyngitis associated with EBV infection. Immunohistochemical analysis identified 5 cases of pharyngotonsillitis associated with herpes simplex virus infection. Serological examination showed that, among 12 cases positive by in situ hybridization, 3 cases were primary infection with infectious mononucleosis and 9 were nonprimary infection. The staining pattern of in situ hybridization was different, ie, a linear pattern in cases of nonprimary infection and a scattered pattern in cases of primary infection. The clinical manifestations of EBV pharyngotonsillitis were distinct from those of herpes simplex virus pharyngotonsillitis and were characteristic irrespective of infectious status, while those of EBV oropharyngitis were more variable. CONCLUSIONS: Epstein-Barr virus-associated pharyngotonsillitis was demonstrated in patients with nonprimary infection unaccompanied by infectious mononucleosis. Epstein-Barr virus should be considered a potential causative agent of oropharyngotonsillitis even in absence of infectious mononucleosis, especially in a young adult. PMID- 10680871 TI - Novel cell proliferation marker for identification of a growth center in the developing human cricoid. AB - BACKGROUND: Developmental histomorphology of the human cricoid cartilage has never been well described. Regional growth centers in the cricoid have been hypothesized, but have never been demonstrated in histological sections. OBJECTIVES: To apply Mib-1 immunostaining, a monoclonal antibody directed at a nuclear proliferation marker, in human cricoids to identify a growth center and to study the changing histomorphology of the developing cricoid. DESIGN: Immunohistochemical Mib-1 studies were performed on postmortem cricoid sections of 2 fetuses (gestational age, 18.5 and 33 weeks), 1 newborn (full term, 41 weeks), and 3 children (aged 1, 4, and 13 years, respectively). Cell counts, surface areas, and organizational patterns of the chondrocytes were studied and described in hemotoxylin-eosin-stained sections. RESULTS: Differential Mib-1 staining was found. The 18.5-week fetus showed diffuse cell proliferation throughout the cricoid. The cricoid sections of the 33-week fetus and 1-year-old child revealed a distinct ring of proliferation in the outer third of the cricoid ring. The 4- and 13-year-old exhibited no cell proliferation. Histomorphologically, with increasing age came chondrocyte hypertrophy, decreasing cell count per standard square, and increasing organization from a scattered to radial columnar pattern. CONCLUSIONS: Growth of the cricoid involves a diffuse pattern of cell proliferation throughout the cricoid in fetal tissue. At term and until age 1 year, the region of proliferation is more restricted to the outer subperichondrial surface. By age 4 years, cell proliferation has stopped. Histomorphologic changes in the developing cricoid include decreasing cell counts per standard unit area, but increasing surface area with age. The aging chondrocytes develop an increasingly organized layout to form a radially arranged columnar pattern similar to that in the growth plate of the developing limb bud. PMID- 10680872 TI - Thirty-four patients with carcinoma of the cervical esophagus treated with chemoradiation therapy. AB - OBJECTIVE: To review the experience of 2 institutions in the management of localized carcinoma of the cervical esophagus with chemoradiation therapy. DESIGN: A series of 34 patients received chemoradiation therapy for a 5-year period. All patients were treated with curative intent. Three different regimens were used, all involving concomitant chemotherapy and high-dose radiation therapy. Data relating to toxic effects, local control of disease, and disease free and overall survival were prospectively collected. SETTING: Two combined clinics at separate major hospitals where multidisciplinary care is the standard practice for this disease. PATIENTS: Patients with biopsy-proved carcinoma of the cervical esophagus. INTERVENTIONS: Patients received 3 different chemotherapy regimens. Two of the regimens used a combination of cisplatin and fluorouracil. The high-dose cisplatin regimen was a large dose of cisplatin (80 mg/m2) given on days 1 and 22 followed by a 96-hour infusion of fluorouracil (800 mg/m2) from days 2 to 5 and from days 23 to 26. The low-dose cisplatin regimen was cisplatin, 20 mg/m2, from days 1 to 5 and from days 22 to 26 and the same 96-hour infusion of fluorouracil. The third regimen used fluorouracil alone. The mean radiation dose administered was 61.2 Gy in 29.6 fractions during 41.8 days using 4- or 6-mV photons and a shrinking field technique. RESULTS: The results of treatment have shown a high rate of local control, although some patients developed metastases. The local complete response rate following treatment was 91%, and the rate of local control of disease was 88%. The projected actuarial 5-year survival rate was 55%. Death from other causes was common. The acute toxic effects of the treatment were acceptable, with only 5 patients requiring nasogastric feeding or gavage. Two patients died of complications related to strictures. CONCLUSION: Concomitant chemoradiation therapy, should be the treatment of choice for carcinoma of the cervical esophagus. PMID- 10680873 TI - Metastases to temporal bones from primary nonsystemic malignant neoplasms. AB - OBJECTIVES: To compare histopathological and clinical findings of metastasis to the temporal bone with previous reports and to determine the prevalence of these metastases in patients with nonsystemic cancer. STUDY DESIGN: Retrospective. METHODS: Autopsy records of 864 patients were screened to select those with primary nondisseminated malignant neoplasms. These were evaluated histopathologically for metastasis to and site of involvement within the temporal bone, and histological characteristics of the tumor. Clinical records and autopsy reports were reviewed for demographic data, clinical course, otologic and vestibular manifestations, site of primary and its histological features, extent of metastasis, and mode of spread. RESULTS: Of 212 patients with primary nondisseminated malignant neoplasms, 47 had metastases to the temporal bone (76 temporal bones). Twenty different primary tumors had metastasized, most commonly breast cancer. Hearing loss was the most common otologic symptom (seen in 19 patients [40%]), while 17 (36%) had no otologic or vestibular symptoms. Temporal bone involvement was bilateral in 29 patients (62%). Most metastases to the temporal bone demonstrated hematogenous spread in 58 temporal bones (76.7%), and petrous apex was the most common site of metastases in 63 temporal bones (82.9%). Temporal bone metastases were not observed in cases where the primary tumor was adequately treated. CONCLUSIONS: In the largest series to date, we found temporal bone metastases more frequently than previously reported. Absence of temporal bone involvement in cases in which the primary tumor was adequately treated stresses the need for early management of cancer. Metastatic disease must be considered as a cause of hearing loss in patients with a history of malignant neoplasm. PMID- 10680874 TI - Radiation and intra-arterial cisplatin: effects on arteries and free tissue transfer. AB - OBJECTIVES: To determine the histopathologic effect of combined intra-arterial cisplatin administration and hyperfractionated external beam radiation treatment (HYPERRADPLAT) on potential recipient arteries in the neck and to analyze the efficacy of free tissue transfer (FTT) in patients undergoing HYPERRADPLAT. DESIGN: Cisplatin-perfused and nonperfused artery segments were harvested during planned interval neck dissection performed 6 to 10 weeks after HYPERRADPLAT. These segments were evaluated by light microscopy and transmission electron microscopy. All patients undergoing FTT after HYPERRADPLAT were reviewed retrospectively. SETTING: Academic medical center. PATIENTS AND INTERVENTION: Eight patients undergoing HYPERRADPLAT for head and neck squamous cell carcinoma and planned interval neck dissection were prospectively studied. All patients had a perfused artery sampled, and 3 also had a nonperfused (control) artery sampled. Five patients undergoing FTT after HYPERRADPLAT were retrospectively analyzed for outcome of FTT. RESULTS: No consistent histological or ultrastructural differences were detected between injected and noninjected arteries. Both demonstrated intimal thickening, collagen and elastin deposition in the intimal layer, and, occasionally, intimal smooth muscle proliferation. A smaller fraction of the injected and noninjected arteries demonstrated smooth muscle cell vacuolation, elastic fiber degeneration, and calcific deposits. Four of 5 FTTs in patients undergoing HYPERRADPLAT were successful. CONCLUSIONS: The changes seen in the injected and noninjected arteries were characteristic of ionizing radiation. Arteries treated with HYPERRADPLAT had no observable difference from vessels treated with radiation alone. These vessels can be used with caution as recipient vessels for FTT. Further clinical experience is needed to establish the expected results of FTT using these arteries. PMID- 10680875 TI - Delayed anterior ischemic optic neuropathy after neck dissection. AB - There are only 2 published cases of anterior ischemic optic neuropathy (AION) occurring after neck dissection, to our knowledge. We report a case of bilateral AION following neck dissection, discuss the differential diagnoses, and compare the features of this case with those of previously described cases. There were none of the previously described risk factors for the development of AION after head and neck surgery (eg, hypotension, facial edema, or sudden massive hemorrhage) in this case, but there was prolonged diffuse postoperative bleeding. Also, the symptoms did not arise before the fifth postoperative day as they did in the other cases. Prolonged, mild postoperative bleeding is a risk factor for AION. Visual loss during the entire first postoperative week has to be regarded as a complication of surgery, requires the exclusion of several differential diagnoses, and must not be confounded with the patient's confusion or symptoms of withdrawal. PMID- 10680876 TI - Bilateral mucosa-associated lymphoid tissue lymphoma of the parotid gland. AB - Mucosa-associated lymphoid tissue (MALT) tumors of the parotid gland are extranodal non-Hodgkin lymphomas. Stage I and II MALT tumors are usually treated with surgery or radiotherapy. Bilateral MALT-derived non-Hodgkin lymphoma of the parotid glands is rare, and optimal treatment is debatable. Two patients presented at the otorhinolaryngology department of the Friedrich-Alexander University of Erlangen-Nuremberg, Erlangen, Germany. The treatment strategy that was used in case 1 was also successfully used in case 2. A precise diagnosis could not be made by either fine-needle biopsy or intraoperative frozen section biopsy; it was achieved with open biopsy. Surgery and/or radiotherapy proved to be effective. There was no recurrence of disease in either case. The advantages of surgery are complete resection of the tumor and absence of xerostomia and mucositis, which are caused by irradiation. Radiotherapy does not produce a scar or an indentation at the parotid region, however, and results in a better cosmetic appearance. Therefore, we recommend open biopsy with facial nerve monitoring and subsequent irradiation in cases in which bilateral prominence of the parotid glands and suspicion of a MALT lymphoma are both present. PMID- 10680877 TI - Laryngeal cleft and eosinophilic gastroenteritis: report of 2 cases. AB - Although laryngotracheoesophageal clefts are often found in association with other well-described anomalies, we know of no previous reported association with eosinophilic gastroenteritis, a disorder of unknown etiology characterized by eosinophilic infiltration of the gastrointestinal tract. We treated 2 children who had laryngeal clefts and eosinophilic gastroenteritis. Since the esophageal inflammatory changes found in eosinophilic gastroenteritis may persist despite aggressive therapy, management of the laryngotracheoesophageal clefts is more complicated. The diagnosis of eosinophilic gastroenteritis should not be overlooked in patients with laryngotracheoesophageal clefts and warrants prompt referral to a pediatric gastroenterologist. PMID- 10680878 TI - Using vision changes to differentiate sinonasal headache from migraine. PMID- 10680879 TI - Visual aura: a useful diagnostic tool. PMID- 10680880 TI - Patients with headache and visual disturbance: a differentiation between migraine and sinonasal headache. PMID- 10680881 TI - Quiz case 1. Cutaneous mucormycosis of the external ear. PMID- 10680882 TI - Quiz case 2. Burkitt lymphoma (BL) of the nasopharynx. PMID- 10680883 TI - Gastric dysplasia: the Padova international classification. AB - A worldwide-accepted histologic, classification of the gastric carcinomatous and precancerous lesions is a prerequisite for a consistent recording of epidemiologic data and for both developing and evaluating primary and secondary preventive efforts. Different nomenclatures have been proposed for gastric precancerous lesions in eastern countries and in Japan. This article presents a classification of gastric precancerous lesions resulting from an international consensus conference involving pathologists of different countries. Five main diagnostic categories are identified. To allow comparisons with the nomenclature proposed by the Japanese Research Society for Gastric Cancer, each category was also assigned a numeric identification: 1 = normal, 2 = indefinite for dysplasia, 3 = noninvasive neoplasia, 4 = suspicious for invasive cancer, and 5 = cancer. The interobserver reproducibility of the histologic classification was tested in a series of 46 cases. By collapsing benign alterations (categories 1+2) versus noninvasive neoplasia (category 3) versus suspicious for invasive cancer and fully appearing carcinomatous lesions (categories 4+5), the general agreement value was 77.7%, whereas kappa coefficient was 0.63. By examining gastric precancerous lesions from diverse populations, the authors agreed that the gastric precancerous process is universal and the differences in nomenclatures are merely semantics. The international Padova classification of the gastric precancerous lesions is submitted to the attention of the international scientific community, which is invited to test and to improve on it. PMID- 10680884 TI - Differentiation of primary and metastatic clear cell tumors in the liver by in situ hybridization for albumin messenger RNA. AB - Clear cell neoplasms presenting as metastatic hepatic masses may be difficult to differentiate histologically and immunohistochemically from hepatocellular carcinoma (HCC) with prominent clear cell features, especially in small biopsy specimens. In situ hybridization (ISH) for albumin messenger RNA (mRNA) has been previously shown to be sensitive and specific for the detection of hepatocellular differentiation, but its use for the identification of clear cell HCC has not been previously evaluated. Among 309 cases of hepatocellular carcinoma diagnosed at Mayo Clinic between 1985 and 1998, 30 cases (9.7%) with at least 30% (range, 30%-90%; median 60%) clear cells were studied by ISH for albumin mRNA. In addition, immunohistochemical expression of AFP and polyclonal CEA, serum determination of AFP, and histopathologic analyses of the tumor were done. Forty two clear cell tumors were used as a control group: 21 metastatic clear cell tumors to the liver (14 renal cell carcinomas and 7 adrenal cortical carcinomas) and 21 primary clear cell tumors of the retroperitoneum (10 renal cell carcinomas, 5 adrenal cortical adenomas, 4 adrenal cortical carcinomas, and 2 ovarian carcinomas). ISH for albumin mRNA was reactive in 28 of 30 cases of clear cell HCC (93%). Clear cell HCC expressed AFP (15 cases; 50%) and polyclonal CEA (19 cases; 63%). Tumors expressed either AFP or polyclonal CEA in 23 cases (77%). Elevated serum AFP was present in 24 of 26 cases (92%). These results indicate that ISH for albumin mRNA is a useful method to distinguish clear cell HCC from other clear cell carcinomas metastatic to the liver and clear cell neoplasms in the retroperitoneum. PMID- 10680885 TI - Relationship and significance of greatest percentage of tumor and perineural invasion on needle biopsy in prostatic adenocarcinoma. AB - Serum prostate-specific antigen (PSA) levels and the biopsy Gleason sum are used along with clinical staging to predict prostatectomy pathology results for men with localized prostate cancer. The additional predictive value of perineural invasion (PNI) in pretreatment prostate needle biopsies for evaluating tumor stage in this setting is controversial. The current study evaluates the independent predictive value of PNI for tumor staging in a cohort of 632 men who underwent radical retropubic prostatectomies for clinically localized adenocarcinoma of the prostate between the years 1994 and 1998. None of these men received hormonal or radiation therapy before surgery. In addition to the Gleason sum, biopsy results contained detailed information regarding tumor burden: 1) total number of biopsy cores involved by adenocarcinoma, 2) greatest percentage of any single biopsy involved by prostate carcinoma (GPC), and 3) total percentage of cancer added over all cores (TPC). The presence or absence of any PNI was recorded. Pretreatment factors were analyzed in a univariate and multivariate fashion to determine their predictive value using the TNM tumor stage (pT2 vs pT3) and the modified tumor staging system, which includes surgical margin status (pT2 vs pT3 or positive surgical margin) as end points. Univariate analysis revealed a significant association between pT3 disease and several preoperative factors including age, Gleason sum, serum PSA, digital rectal examination, PNI, GPC, TPC, and the total number of positive cores (p <0.01). Multivariate analysis indicated that serum PSA, Gleason sum, age, and GPC contributed significantly to predicting pT3 disease with odds ratios of 2.7 (95% CI, 1.7-4.3), 2.3 (95% CI, 1.7-3.1), 1.7 (95% CI, 1.1-2.7), and 1.7 (95% CI, 1.4 2.1) respectively. PNI was significant in multivariate analysis only when GPC and TPC were not considered, due to a significant interaction between GPC and PNI (p <0.0001, Wilcoxon's rank sum test). These predictive factors showed a similar relationship to adverse pathology when an alternative definition of adverse pathology was used that included positive surgical margins (pT3 or any positive margin). In the interaction between GPC and PNI, GPC was more significant than PNI in predicting pT3 disease. However, PNI added additional information when adverse pathology was defined more broadly as pT3 or any positive margin. PMID- 10680886 TI - Epithelial sheath neuroma: a new entity. AB - The authors describe four examples of a peculiar cutaneous lesion characterized histopathologically by a proliferation of enlarged nerve fibers ensheathed by squamous epithelium involving the superficial dermis. The perineural epithelial sheaths were composed of uniform squamous epithelium with evidence of cornification in the form of dyskeratotic cells or resulting in orthokeratotic basket-weave corneocytes. Immunohistochemical studies confirmed the epithelial and neural nature of the two components of the lesions, with the nerve fibers expressing immunoreactivity for S-100 protein, neurofilaments, CD57, and nerve growth factor receptor, whereas the perineural epithelial sheaths showed immunoreactivity for cytokeratins. The authors propose the term "epithelial sheath neuroma" for this lesion and believe that it is a distinct and a previously undescribed benign neoplasm of both cutaneous nerves and epithelial elements. PMID- 10680887 TI - Large, central acellular zones indicating myoepithelial tumor differentiation in high-grade invasive ductal carcinomas as markers of predisposition to lung and brain metastases. AB - High-grade invasive ductal carcinomas (IDCs) of the breast with large, central acellular zones on their cut surfaces are usually associated with the myoepithelial immunophenotype of carcinoma cells, which includes the expression of S-100 protein, alpha-smooth muscle actin, and keratin 14. To clarify the clinical significance of these features of IDCs, the authors compared the incidence of the myoepithelial immunophenotype immunohistochemically, patient prognosis, and metastatic sites of the tumor between 20 high-grade IDCs with large, central acellular zones and 40 control high-grade IDCs without these zones. The myoepithelial immunophenotype was detected in 16 IDCs (80%) with large, central acellular zones but in only seven IDCs (18%) without. The risk ratio of metastasis, especially in the brain and lung, and death from cancer were significantly higher (p = 0.0096 and p = 0.030) for the 20 IDCs with large, central acellular zones than for those without by Cox's univariate analysis. Using Cox's multivariate analysis, large, central acellular zones in IDCs were an indicator of high risk of brain and lung metastases and of death by cancer independent of nodal status and tumor size. Examination of large, central acellular zones and myoepithelial immunophenotype in high-grade IDCs appears helpful in predicting patient prognosis and preferential metastatic sites of the tumors. PMID- 10680888 TI - Use of antibodies to RCC and CD10 in the differential diagnosis of renal neoplasms. AB - The majority of renal neoplasms can be distinguished on the basis of histologic examination alone; however, there are morphologic similarities between clear cell renal carcinoma and chromophobe cell carcinoma, as well as between the granular/eosinophilic variants of these tumors and renal oncocytoma. Only a limited number of histochemical markers are available to aid in the differential diagnosis of these neoplasms. Hale's colloidal iron usually yields strong, diffuse cytoplasmic staining of chromophobe cell carcinomas whereas clear cell carcinomas are generally negative; however, interpretation of this stain is not always straightforward. By immunohistochemistry, vimentin is detectable in most clear cell carcinomas and is absent from most chromophobe cell tumors and oncocytomas, but reliance on a single antibody can be misleading. In this report we examine the use of commercially available monoclonal antibodies to RCC and CD10 in the differential diagnosis of common renal tumors. Eighty-five percent of clear cell carcinomas (53 of 62) had detectable surface membrane staining for RCC, and 94% (58 of 62) were positive for CD10. Papillary carcinomas were likewise strongly positive for RCC and CD10 in nearly all cases (13 of 14 each). In contrast, all 19 chromophobe cell carcinomas examined were completely negative for surface membrane staining with both of these markers. Oncocytomas were also negative for RCC (0 of 9), but CD10 was detectable in some cases (3 of 9). These results suggest that the presence of surface membrane staining for RCC and CD10 may be used to confirm a diagnosis of suspected clear cell or papillary renal carcinoma. Chromophobe cell carcinomas should be negative for both markers. The absence of RCC staining may also be helpful in the diagnosis of renal oncocytoma. PMID- 10680889 TI - Esophageal stromal tumors: a clinicopathologic, immunohistochemical, and molecular genetic study of 17 cases and comparison with esophageal leiomyomas and leiomyosarcomas. AB - Although rare elsewhere in the gastrointestinal tract, leiomyomas (LMs) are the most common esophageal mesenchymal neoplasms. In contrast, gastrointestinal stromal tumors (GISTs) predominate in the stomach and intestines but have not been documented in the esophagus. This study was undertaken to determine the clinicopathologic features and frequency of esophageal GISTs compared with LMs and leiomyosarcomas (LMSs) of the esophagus. A total of 68 stromal/smooth muscle tumors from the Armed Forces Institute of Pathology and the Haartman Institute of University of Helsinki were reclassified by current histologic and immunohistochemical criteria. There were 17 GISTs, 48 LMs, and three LMSs. The esophageal GISTs occurred in 12 men and five women with a median age of 63 years (range, 49-75 years). All tumors were from the lowest third of the esophagus, and the most common complaint was dysphagia, whereas two tumors were detected incidentally. Histologically the tumors had an overall basophilic appearance and showed combinations of solid, myxoid, and perivascular collarlike patterns with a spindle cell histology in 13 patients and epithelioid histology in four patients. All tumors were positive for CD117 and for CD34, whereas two patients were also positive for alpha-smooth muscle actin (SMA) and three patients were positive for desmin. One patient showed a unique immunophenotype with coexpression of CD117, CD34, SMA, and desmin. Nine patients died of disease, including all who had a tumor larger than 10 cm, and also one patient whose tumor showed five mitoses per 50 high-power fields. In comparison, esophageal LMs (n = 48) occurred in a younger population (median age, 35 years) but, similar to the GIST group, men predominated (67%). All LMs were clinically indolent tumors with no tumor-related mortality. The LMs showed eosinophilic cytoplasm, and were positive for desmin and SMA, and negative for CD117 and CD34. All three LMSs were large high-grade tumors that showed muscle cell markers but no CD117. All patients died of disease. Esophageal GISTs showed mutations in exon 11 of c-kit as described previously in gastric and intestinal GISTs. The separation of GISTs from esophageal LMs is important diagnostically because the former group has a high risk of malignant behavior. PMID- 10680890 TI - Small cell neuroendocrine carcinoma with skeletal muscle differentiation: report of three cases. AB - Three cases of neuroendocrine carcinoma showing skeletal muscle differentiation are presented. The tumors were located in the skin and subcutaneous tissue, the urinary bladder, and the nasal cavity respectively, and were composed by two cell types admixed intimately with each other. One cell type had features identical to those seen in conventional small cell neuroendocrine carcinoma, including scanty cytoplasm, round nuclei with fine granular chromatin, immunohistochemical reactivity for neuron-specific enolase, chromogranin and cytokeratins, and electron-dense granules on ultrastructural examination. The second cell type was either plasmacytoid or elongated and straplike, with abundant eosinophilic cytoplasm and irregular nuclei with prominent nucleoli. These cells showed immunohistochemical positivity for desmin, sarcomeric actin, myoglobin, and myogenin. They also exhibited ultrastructural evidence of rhabdomyoblastic differentiation in the form of contractile filaments with abortive Z-band formation. An origin from a cell capable of dual differentiation toward neuroendocrine and rhabdomyoblastic elements is postulated for these tumors. PMID- 10680891 TI - Cellular pseudosarcomatous fibroepithelial stromal polyps of the lower female genital tract: an underrecognized lesion often misdiagnosed as sarcoma. AB - Fibroepithelial stromal polyps of the vulvovaginal region are benign lesions that, when bland or hypocellular, are readily recognized. However those that exhibit bizarre cytomorphology, atypical mitoses, or hypercellularity, raising the possibility of malignancy, continue to be underrecognized. The authors reviewed a series of fibroepithelial stromal polyps to characterize further the morphologic features that can lead to a misdiagnosis of sarcoma. A total of 33 of 65 consecutive cases of fibroepithelial stromal polyps retrieved from the authors' consultation files were remarkable for marked hypercellularity (33 of 33), marked cytologic pleomorphism (21 of 33), mitotic counts of more than 10 mitoses per 10 high-power fields (12 of 33), and the presence of atypical mitoses (14 of 33). A total of 16 of 33 lesions had three or more of these features. Important morphologic clues to the diagnosis (shared with usual polyps at this site) were lack of an identifiable lesional margin, extension of abnormal stromal tissue up to the mucosal-submucosal interface, and the frequent presence of individually scattered multinucleate stromal cells, most often located close to the surface epithelium. Immunohistochemically, seven of 12 cases were desmin positive and one of 11 cases were smooth muscle actin positive. The age range of patients was 16 to 75 years (median, 32 years), and 21 patients (64%) were premenopausal. Sites included the vagina (18 of 33), cervix (seven of 33), and vulva (eight of 33). A total of 14 of 33 patients were pregnant, three patients were taking Tamoxifen, and one patient was on oral progesterone. Eight of 33 patients had multiple lesions at the time of presentation, of whom five were pregnant. Clinical follow-up was available in 21 of 33 patients. Three of 21 patients with follow-up had local, nondestructive recurrence. Two of these patients had multiple recurrences. None of the patients followed developed metastases. Cytologic atypia has been a previously recognized feature in these lesions; however, the occurrence of marked stromal cellularity and a mitotic rate of more than 10 mitoses per 10 high-power fields have not been emphasized previously. Moreover, the combination of these features has only rarely been documented. Awareness of the spectrum of histologic features that these lesions can exhibit is crucial in their accurate recognition, thus avoiding potential overtreatment. PMID- 10680892 TI - Independent prognostic value of peritoneal immunocytodiagnosis in endometrial carcinoma. AB - Among the clinical parameters that play a pivotal role in predicting the outcome of patients with endometrial carcinoma, intraperitoneal microscopic dissemination represents an important cause of recurrences. To date, peritoneal cytology has been incorporated into the current surgical staging system (International Federation of Gynecology and Obstetrics 88), although its predictive value remains a controversial issue. In this study the authors investigated the possibility of applying immunocytochemistry (ICC) to the diagnosis of peritoneal washing (PW) aimed at improving conventional cytology and verifying the prognostic value of peritoneal malignant cells. The authors analyzed 182 PWs sampled from endometrial cancer patients. The ICC analysis was performed using two monoclonal antibodies (MAbs)--AR-3 and B72.3--that in combination recognize more than 95% of endometrial carcinomas. The presence of peritoneal-free cancer cells was identified morphologically in 27 of 182 lavages (14.8%) and ICC in 50 of 182 (27.5%), with a significant improvement (p <0.0001). Five-year survival analysis, comparing results of ICC and cytodiagnosis, demonstrated a significant decrease of disease-free survival in patients with peritoneal microscopic disease. Furthermore, multivariate analysis showed that ICC diagnosis of PWs is an independent prognostic factor. Data indicate that the use of selected MAbs allows one to identify cytologically false-negative cases, providing results that are highly predictive of a worse clinical outcome. PMID- 10680893 TI - Primary giant cell tumor of soft tissues: a study of 22 cases. AB - Twenty-two cases of giant cell tumor of soft tissues (GCT-ST) identified in the Mayo Clinic files and the consultation files of two of the authors (A.G.N., C.D.M.F.) were analyzed clinicopathologically. Age at presentation ranged from 5 to 80 years (median, 43 years), and there was no sex predilection (12 male, 10 female). Duration of symptoms ranged from 2 to 12 months (median, 4.5 months), and a painless growing mass was the most common complaint. The lower limbs were the most frequent location (50%), followed by the trunk (31.8%) and the upper limbs (13.6%). The size of the tumors ranged from 1 to 10 cm, and they tended to be superficial (86.4%), forming well-circumscribed (72.7%), multinodular (86.4%) masses. Histologically, all tumors consisted of a mixture of mononuclear cells showing vesicular, round to oval nuclei and osteoclastlike, multinucleated giant cells distributed uniformly throughout the tumors. Foci of stromal hemorrhage were observed in 11 tumors (50%); nine tumors (40.1%) showed metaplastic bone formation and six (27.2%) showed aneurysmal bone cystlike areas. Necrosis was absent in all but one tumor. Mitotic figures were present in all but one tumor, ranging from two to more than 30 mitoses per 10 high-power fields (HPFs; median, 9.5 mitoses per 10 HPFs) and were typical in aspect. Vascular invasion was identified in seven tumors (31.8%), and none of the tumors showed marked cellular atypia or pleomorphism. The tumors were treated surgically, and follow-up information was available for 16 patients (duration of follow-up, 2 to 130 months; median, 51 months). Only one of the 16 patients (6.2%) had local recurrence and lung metastases; this patient died of the tumor. In conclusion, GCT-ST occurs as a primary soft-tissue neoplasm and is identical clinically and morphologically to giant cell tumor of bone. Provided that GCT-ST is treated adequately by complete excision, a benign clinical course is expected because episodes of distant metastasis and tumor-associated death seem to be exceedingly rare. PMID- 10680894 TI - The pathology of late recurrence of testicular germ cell tumors. AB - A total of 91 men had histologically documented late recurrences of testicular germ cell tumors characterized by a complete response to treatment with a subsequent disease-free interval of at least 2 years and no evidence of a second primary lesion. Ninety percent of the patients for whom information was available received chemotherapy shortly after their initial diagnosis of testicular germ cell tumors; most of the other patients were known to have stage I disease initially. Overall, 60% of patients had teratoma in their late recurrences, including 20 patients (22%) in whom teratoma was the only element. Thus, teratoma was the most common type of neoplasm in late recurrences. Excluding teratoma coexisting with other types of neoplasms, yolk sac tumor was the most frequent type of tumor in patients with late recurrence. It occurred in 47% of patients, either alone or with teratoma, another nonteratomatous germ cell tumor type, or a "nongerm cell malignant tumor." Unusual types of yolk sac tumor, including glandular, parietal, clear cell, and pleomorphic patterns, were seen frequently in late recurrences and often raised differential diagnostic problems with "nongerm cell" carcinomas. A smaller number of late recurrences consisted of other types of neoplasms. Twenty percent of patients with late recurrence had a nonteratomatous germ cell tumor other than yolk sac tumor, either alone, with yolk sac tumor, or with a "nongerm cell malignant tumor." Most of these nonteratomatous germ cell tumors other than yolk sac tumor were embryonal carcinoma, although rarely seminoma and choriocarcinoma were encountered. "Nongerm cell malignant tumors," including both sarcomas and carcinomas of various types, occurred in 23% of late-recurrence patients, either alone or with a nonteratomatous germ cell tumor. Late recurrences were seen in many different sites in these patients, including the retroperitoneum, abdomen, pelvis, liver, mediastinum, lung, bone (femur, vertebra, and rib), lymph nodes outside the retroperitoneum and mediastinum (supraclavicular, neck, and axillary regions), scrotum and inguinal regions, adrenal gland, chest wall, and buttocks. Follow-up data were available for 79 of the 91 patients studied. Duration of follow-up ranged from 2 months to 13 years after the patient's first late recurrences; the mean length of follow-up was 4.8 years. Patients whose late recurrences consisted of teratoma only had the most favorable outcomes, with 79% having no evidence of disease at last follow-up. Patients whose late recurrences consisted of pure "nongerm cell malignant tumor" or pure germ cell tumor (yolk sac tumor or other types) had a much worse prognosis: Only 36% to 37% were alive with no evidence of disease. Patients with two different types of nonteratomatous malignancies in their late recurrences had a dismal clinical course: Only 17% with both yolk sac tumor and other nonteratomatous germ cell tumor had no evidence of disease, whereas no patient with both nonteratomatous germ cell tumor and "nongerm cell malignant tumor" was disease free. Late recurrences consisting of teratoma alone often have a favorable outcome, but the prognosis in all other patients is poor. Furthermore, late recurrence is not likely to respond to chemotherapy and is best treated by surgical excision when possible. PMID- 10680895 TI - Immunohistochemically detected micrometastases in peribronchial and mediastinal lymph nodes from patients with T1, N0, M0 pulmonary adenocarcinomas. AB - The T1, N0, M0 subset of stage I lung adenocarcinoma is a tumor that has a 5-year disease-free survival rate of 66% to 85%. To date, there has not been a rigorous immunohistochemically detected lymph node micrometastasis study composed of patients with identical stage and type of tumors, and in which standard histologic features were incorporated into multivariate analyses. We immunohistochemically examined the peribronchial and mediastinal lymph nodes from 80 consecutively accrued patients with T1, N0, M0 adenocarcinomas and bronchioloalveolar carcinomas unselected for distant metastasis, and an additional 39 patients with similar stage and type neoplasms who were selected for their development of metastases to evaluate the prevalence of micrometastases, their association with distant metastases, and their relationship with other pathologic prognostic features. All slides were stained with keratin AE1/3. Micrometastases were confirmed with Ber-Ep4. Three immunohistochemically detected lymph node micrometastases were identified in three of 80 consecutively accrued patients (4%). These three positive stains constituted 0.5% of the 573 stains required to immunohistochemically screen all of the lymph node blocks from these patients. Among the 39 patients who were selected because they developed distant metastases, three immunohistochemically detected lymph node micrometastases from three patients were identified, which constituted 8% of patients in this group and 1% of the 280 stains required to screen all of these patients' lymph nodes. Small vessel invasion, maximum tumor dimension, and immunohistochemically detected lymph node micrometastases were independently associated with metastases on multivariate analysis. Among patients who developed metastases, there was no significant difference in the disease-free survival rate between those with and those without immunohistochemically detected lymph node micrometastases. Given the low sensitivity in terms of the number of immunohistochemical stains performed, and the prognostic significance of standard histologic features, the use of immunohistochemical screening lymph nodes from all patients with T1, N0, M0 adenocarcinomas is questionable. PMID- 10680896 TI - Prevalence of inter-institutional anatomic pathology slide review: a survey of current practice. AB - Multiple studies have demonstrated discrepancy rates between original and review histopathologic diagnoses of up to 30% with a mean of approximately 10%. In view of these rates of discrepancy, several authorities, including the Association of Directors of Anatomic and Surgical Pathology, have recommended in-house review of all outside materials before commencement of therapy. We used a mail survey to determine the degree of compliance with these recommendations among pathology groups in the United States. Mail surveys were sent to six randomly selected hospitals from each state (300 total). The survey included demographic questions, including surgical pathology caseload, size of hospital (beds), and type of hospital (community-general, non-academic-tertiary care, or academic-tertiary care). The survey asked whether the hospital required review of all outside slides before the performance of surgery. If not, was such a policy encouraged but not required. The survey also asked whether in-house review of outside cases had disclosed any significant differences in pathologic diagnoses. Finally, the survey questioned whether any discrepancies between an internal and external surgical pathology diagnosis had been discovered following radical surgery. One hundred twenty-six usable responses were obtained. Fifty-five of these were from hospitals self-described as community-general, seven were from hospitals describing themselves as non-academic-tertiary care, and the remaining 61 hospitals described themselves as academic-tertiary care institutions. Sixty three institutions stated they had a requirement for in-house review of outside material, with 46 of 61 academic-tertiary centers having such a requirement. Thirty-seven of 55 community-general hospitals did not require in-house review of outside material before surgery could be performed. One hundred ten of the 126 institutions returning surveys either encouraged or required review of outside material. Ninety-five institutions reported that they had at least one outside case in which their diagnosis was significantly discordant with that rendered by the referring pathologist. Sixty (48%) of the 126 institutions reported at least one case in which a discrepancy was found between the outside biopsy diagnosis and the internal diagnosis rendered on material obtained by radical surgery. Approximately half of all responding institutions have a requirement for in-house review of outside material prior to surgery. A majority of institutions requiring such review have found discrepancies between the in-house diagnoses and those rendered by referring laboratories. PMID- 10680898 TI - Ronald A. DeLellis, M.D., recipient of the 1999 Fred W. Stewart Award. PMID- 10680897 TI - Mesotheliomas with deciduoid morphology: a morphologic spectrum and a variant not confined to young females. AB - Deciduoid mesotheliomas are rare with only four previously reported cases, all affecting the peritoneum of young females. We describe another six cases (three men and three women; age range 52-65 yrs, median 55 yrs; five peritoneal and one pleural). Three patients had an occupational history of asbestos exposure. The deciduoid appearance predominated in four cases, whereas in two it represented a minor component within conventional tubulopapillary epithelioid mesothelioma. All tumors were strongly cytokeratin-positive (including CK5/6) and all showed at least focal staining for thrombomodulin, HBME-1, and calretinin. All were negative for epithelial mucin (D/PAS), CEA, BerEP4, LeuM1 (CD15), CD21, CD35, and S100 protein. Five of six cases (83%) were vimentin-positive and two (33%) were focally positive for alpha-smooth muscle actin. A differential diagnosis of gastrointestinal autonomic nerve tumor (GANT) had been initially considered from the morphology of one case, and we found positivity for some of the "neuronal" markers described in GANTs. This prompted us to apply such a panel to the other five tumors, accepting that the cytokeratin positivity encountered in all of our cases would exclude GANT. All cases of deciduoid mesothelioma (100%) were positive for PGP 9.5 and NSE and four of six (66%) were positive for NKI/C3. Weak focal staining (<5% cells) for synaptophysin was seen in two of six tumors. All cases were chromogranin-negative. All cases examined by electron microscopy showed desmosomes and smooth microvilli without rootlets but no neuroendocrine granules. In conclusion, a deciduoid morphology appears to be part of the histopathologic spectrum encountered in epithelioid mesothelioma. This variant is not confined to female patients and occurs over a wider age range than previously recognized. The overlapping immunophenotype with GANTs illustrates that caution should be exercised when interpreting positivity for "neuronal" markers in this context. An immunohistochemical panel that includes cytokeratins should always be used. PMID- 10680899 TI - Spontaneously relapsing clonal, mucosal cytotoxic T-cell lymphoproliferative disorder: case report and review of the literature. AB - Primary T-cell lymphoma of the gastrointestinal tract is a rare and usually aggressive disorder that may be associated with celiac disease. The authors describe a unique case of a clonal proliferation of CD8+ T cells involving the oral mucosa, ileum, and colon of a 35-year-old man that has regressed spontaneously and recurred numerous times over a 9-year period without treatment. The patient's symptoms were limited to occasional rectal bleeding and recurring painful oral ulcers. Within the intestine, these collections of small T cells induced minimal architectural distortions and did not show extensive epitheliotrophism. Polymerase chain reaction and sequencing analyses revealed that the identical T-cell clone has been present for more than 9 years and in different mucosal locations in this patient. This may represent a unique T-cell lymphoproliferative process akin to a mucosal counterpart of lymphomatoid papulosis of the skin. PMID- 10680900 TI - 'Little' littoral cell angioma of the spleen. PMID- 10680901 TI - Examining the sentinel lymph node. PMID- 10680902 TI - Epithelial-myoepithelial carcinoma. PMID- 10680903 TI - Epstein-Barr virus-related intravascular lymphomatosis. PMID- 10680904 TI - Poorly differentiated thyroid carcinoma--it is important. PMID- 10680905 TI - Adenoid basal epithelioma versus adenoid basal carcinoma. PMID- 10680906 TI - Nodal marginal zone lymphoma. PMID- 10680907 TI - Primary nodal marginal zone lymphomas of splenic and MALT type. PMID- 10680908 TI - What is a neuroendocrine tumor? PMID- 10680909 TI - Omit iodine and CD30 will shine: a simple technical procedure to demonstrate the CD30 antigen on B5-fixed material. PMID- 10680910 TI - 'Like--but oh, how different!'. PMID- 10680911 TI - Lobular endocervical glandular hyperplasia represents pyloric gland metaplasia? PMID- 10680912 TI - Lobular endocervical glandular hyperplasia represents pyloric gland metaplasia? PMID- 10680913 TI - Cause of familial and multiple gastrointestinal autonomic nerve tumors with hyperplasia of interstitial cells of Cajal is germline mutation of the c-kit gene. PMID- 10680914 TI - Controversies in the management of thyroid nodule. AB - Few subjects in surgery have generated as much controversy as the management of thyroid nodule. The controversial issues include classification and histology, diagnostic evaluation including needle biopsy, indications for surgery, management of incidentalomas of the thyroid, the role of frozen section, extent of thyroidectomy, management of neck nodes, the role of suppressive therapy, the use of radioactive iodine, and appropriate follow-up. The two major issues in relation to the controversies are diagnostic workup and extent of thyroidectomy. Whenever the issue related to extent of thyroidectomy is discussed, there are two strong groups believing in total thyroidectomy or less than total thyroidectomy. This has generated considerable debate and panel discussions, and this article reviews this on-going debate. PMID- 10680915 TI - Prevalence of herpes simplex virus in malignant laryngeal lesions. AB - OBJECTIVE: To determine the prevalence of herpes simplex virus (HSV) in malignant laryngeal lesions. STUDY DESIGN: Retrospective review. MATERIALS AND METHODS: Paraffin-embedded, histologically confirmed specimens containing benign laryngeal lesions, squamous cell carcinoma of the larynx, and squamous cell carcinoma of the oral cavity were identified from archived surgical specimens. Biopsies of normal-appearing oral cavity tissue were also obtained from fresh-frozen cadavers. These tissues were analyzed for the presence of HSV DNA using polymerase chain reaction techniques. Patient charts were reviewed for patient demographics, risk factors, stage, clinical course, treatment, and outcome. RESULTS: HSV was detected in nine laryngeal squamous cell carcinomas (75%) and in none of the benign laryngeal lesions (P = .0001). HSV was also found in three oral cavity squamous cell carcinomas (25%) and in none of the controls (P = .049). CONCLUSION: HSV is more prevalent in squamous cell carcinoma of the larynx and oral cavity than in their respective control groups, suggesting a role for carcinogenesis. HSV is more prevalent in squamous cell carcinoma of the larynx than of the oral cavity. Further studies to determine the role of HSV as a cocarcinogen and its interrelationship with other environmental factors in laryngeal cancer are warranted. PMID- 10680916 TI - Sentinel lymph node radiolocalization in head and neck squamous cell carcinoma. AB - OBJECTIVES: To determine the feasibility of sentinel node radiolocalization in stage N0 in head and neck squamous cell carcinoma and to gain insight as to whether the sentinel node could be prognostic of regional micrometastatic disease. STUDY DESIGN: A prospective report on the application sentinel node radiolocalization in eight patients with N0 squamous cell carcinoma of the head and neck region. METHODS: For each patient a peritumoral submucosal injection of filtered technetium (99mTc) prepared with sulfur colloid was performed immediately following intubation. After at least 30 minutes, focal areas of accumulation corresponding to a sentinel node were marked on the skin surface. Complete neck dissections were performed, and the sentinel nodes were identified for later histological evaluation and comparison to the remaining lymphadenectomy specimen. RESULTS: Sentinel node radiolocalization accurately identified two or more sentinel lymph nodes in all eight cases. In one patient, two of the three lymph nodes containing micrometastatic disease were sentinel lymph nodes. There was no instance in which sentinel node was negative for micrometastatic disease while being positive in a nonsentinel lymph node. CONCLUSIONS: Accurate localization of the sentinel lymph node using radiolabeled sulfur-colloid is feasible in patients with squamous cell carcinoma of the head and neck region. Although sentinel node radiolocalization in head and neck squamous cell cancer may potentially reduce the time, cost, and morbidity of regional lymph node management, more experience with technique is required before its role can be determined. PMID- 10680917 TI - Long-standing lateral neck mass as the initial manifestation of well differentiated thyroid carcinoma. AB - OBJECTIVE: To analyze the presentation, evaluation, and treatment of a subset of patients with well-differentiated thyroid carcinoma who present with a lateral neck mass and no palpable disease in the thyroid gland. STUDY DESIGN: A retrospective review of all patients undergoing thyroidectomy for malignancy. METHODS: A database of all thyroidectomies performed for malignancy by the Vanderbilt University Department of Otolaryngology-Head and Neck Surgery from 1992 to 1997 was created. Patients who presented with an isolated neck mass without evidence of palpable disease in the thyroid were selected for the study population. RESULTS: There were 60 cases of thyroid malignancy, with 14 cases (23.3%) that presented as isolated lateral neck mass. The characteristics of this group (compared with the population of all thyroid malignancies) include younger age at presentation (37.7 +/- 15.2 y vs. 49.8 +/- 15.6 y; Student t test: P = .019) and long-standing presence of symptoms (27.4 +/- 39.6 mo vs. 3.6 +/- 3.9 mo; P = .023). These patients generally presented from a referring facility after having an excisional biopsy, which was 100% accurate. Fine-needle aspiration is becoming more useful and was 66.7% accurate. Histological examination revealed cancer in the thyroid gland in all patients, 11 cases of papillary carcinoma, 2 follicular carcinomas, and one medullary carcinoma The mean size of the primary focus was 10.9 +/- 8.7 mm, with 29% demonstrating bilateral disease and 14% demonstrating multifocal disease in the ipsilateral gland. The neck specimens revealed an average of 5.3 +/- 3.2 metastatic nodes in levels II-IV and 3.9 +/- 4.6 metastatic nodes in the paratracheal region. CONCLUSION: Based on this patient population, the long-standing lateral neck mass in the young patient should raise the physician's index of suspicion for thyroid carcinoma Fine-needle aspiration should be used in conjunction with judicious excisional biopsy. The bilateral and multifocal nature of otherwise occult primary disease argues for total thyroidectomy in this setting. PMID- 10680918 TI - Length of stay after free flap reconstruction of the head and neck. AB - OBJECTIVES: To analyze the incidence and timing of postoperative complications after free tissue transfer (FTT) and relate that to length of stay (LOS.) STUDY DESIGN: We reviewed one surgeon's experience with 97 patients undergoing 100 head and neck reconstructions via FTT for a variety of traumatic and ablative defects METHODS: Charts were reviewed for demographic data, type of defect and flap, complications, LOS, length of intensive care unit (ICU) stay, date of decannulation, and first oral intake, any readmission to the hospital, and preoperative radiation status. RESULTS: Using strict guidelines, 31% of patients had some form of complication, including a 9% flap failure rate. Average postoperative LOS for all patients was 11 days. Average LOS for uncomplicated cases was 9 and for complicated cases was 16 days. For cases with flap-related complications the average LOS rose to 20 days. All reconstructive failures (defined as patients requiring subsequent surgical procedures after a flap related complication, regardless of outcome) occurred within the first 7 postoperative days. Three patients were readmitted for various reasons: a partial flap dehiscence (postoperative day [POD] 9), meningitis (POD 24), and orocutaneous fistula (POD 22), for a 3.2% readmission rate. Fourteen percent of patients were on a regimen of oral intake, and 13% had decannulation by the time of discharge. Resumption of oral intake and tracheostomy decannulation were accomplished on an outpatient basis in the remainder of patients. CONCLUSIONS: There were no preventable complications associated with early hospital discharge, nor was there evidence of adverse patient outcome. We conclude that early hospital discharge is feasible after FTT reconstruction and is consistent with quality care. PMID- 10680919 TI - Radiation dose to otologic structures during head and neck cancer radiation therapy. AB - BACKGROUND: Otologic structures are often contained within head and neck cancer radiation treatment ports. The dosimetry to otologic structures has not been routinely analyzed and radiation treatment planning does not currently attempt to specifically avoid the inner ear structures when dosimetry is calculated. Recent studies demonstrate that up to 30% of patients experience sensorineural hearing loss on multimodality therapy with cisplatin and radiation. METHODS: In the current case series, radiation dosimetry to otologic structures was calculated from computed tomogram treatment plans on patients. Fifteen nasopharyngeal, oral cavity, oropharyngeal, and hypopharyngeal cancer patients were analyzed. RESULTS: Between 8% and 102% of the total dose is delivered to the petrous bone/cochlea, with 4 of 15 patients getting more than 50% of the dose to at least one cochlea The mastoid air cells received between 3% and 75% of the total dose, with higher doses being delivered to patients with bulky high neck metastases or nasopharyngeal tumors. The eustachian tubes received between 20% and 102% of the total dose, with 10 of 15 patients receiving more than 50% of the dose to this anatomic site. CONCLUSION: We conclude that the cochlea and eustachian tubes receive significant radiation during treatment, particularly in nasopharyngeal cancer patients. Careful design of radiation treatment ports may allow for the reduction of radiation to hearing structures. PMID- 10680920 TI - Quantification of the learning curve for percutaneous dilatational tracheotomy. AB - OBJECTIVES/HYPOTHESIS: Although numerous investigators have reported a bedside percutaneous dilatational tracheotomy (PDT) complication incidence similar to that of standard operative tracheostomy, others have proposed a "learning curve" for PDT resulting in increased complications early in individual or institutional experience with this procedure. The objective of this investigation is to characterize and quantify the proposed learning curve for PDT. STUDY DESIGN: Prospective analysis of complication incidence for the first 100 PDT procedures performed in a local community hospital Department of General Surgery. METHODS: Demographic data, patient disease variables, and patient anatomic features, as well as perioperative, postoperative, and late complications, were recorded prospectively. Patients were divided into sequential cohorts of 20 and were evaluated for complications at regular intervals. RESULTS: Perioperative and late complication incidence was significantly higher in the first 20 patients who underwent PDT. However, postoperative complication incidence did not significantly vary with operator or institutional experience. In addition, patients with suboptimal anatomy were found to have a significantly increased complication incidence, independent of operator and institutional experience. CONCLUSIONS: Percutaneous dilational tracheotomy has an identifiable learning curve that is most prominent in the first 20 patients treated. Early experience with PDT should be obtained under controlled circumstances, ideally the operating suite. Although most complications occur during acquisition of early experience with PDT, certain life-threatening complications such as tube dislodgment or inability to complete procedure may occur even after extensive experience is obtained. Bedside PDT has an acceptable complication incidence, but any surgeon employing this technique must be prepared to perform immediate standard open tracheotomy to minimize potentially lethal complications of this elective procedure. PMID- 10680921 TI - Documentation of variations in sinonasal anatomy by intraoperative nasal endoscopy. AB - OBJECTIVES: Functional endoscopic sinus surgery (FESS) requires a thorough understanding of the variability in sinonasal anatomy. Previous reports have relied primarily on anatomic studies of cadaveric specimens or skulls, or on radiographic analysis. Relatively few comparative anatomic data have been accumulated with endoscopic examination of living patients. STUDY DESIGN: Retrospective review of video recordings of 119 consecutive patients undergoing intraoperative nasal endoscopy at the time of sinonasal surgery. METHODS: At the beginning of each surgical procedure, endoscopic examination of the nasal cavities was performed with 0 degrees and 30 degrees telescopes and recorded with a three-chip video camera on 3/4-inch U-matic videotape. These video records were then reviewed with attention to variations in anatomical configuration of different sinonasal structures. RESULTS: Data demonstrating variations in the anatomical configuration of the following structures of the lateral nasal wall are presented. Middle turbinate: typical (63%), concha bullosa (15%), sagittal cleft (6%), laterally displaced (4%), "L" shaped (3%), medially bent (3%), laterally bent (3%), medially displaced (2%), and transverse cleft (0.5%). Uncinate process: typical (85%) and medially rotated (15%). Ethmoid bulla: typical or balloon (45%), sausage-shaped (34%), and flat (21%). Accessory ostium: round (50%), oval (46%), and kidney-shaped (4%). Sphenoid sinus ostium: oval (42%), slit (32%), and round (26%). The classification system for the anatomical categories is illustrated with digitized images. CONCLUSIONS: This study attempts to provide statistical data regarding variations in sinonasal anatomy in living subjects. Familiarity with such anatomy is important in differentiating normal variants from pathological conditions to optimize surgical treatment of sinus disease, while avoiding complications. PMID- 10680922 TI - Update on intratympanic gentamicin for Meniere's disease. AB - OBJECTIVE: To review the literature relating to intratympanic gentamicin injection therapy for Meniere's disease to detect consistencies and differences that might suggest optimal technique. DESIGN: Retrospective literature review. METHODS: Eighteen papers from the literature regarding clinical experience with intratympanic gentamicin injections for treatment of Meniere's disease were reviewed and tabulated. RESULTS: All papers reported high success rates in treating episodic vertigo of Meniere's disease, but technique, dose, duration, and treatment philosophy varied considerably. Hearing loss was typically reported in about 30% of patients. CONCLUSION: No single technique of gentamicin injection has a significant medical advantage over the others. Until controlled studies indicate otherwise, nonmedical needs such as convenience and safety may be considered when choosing a technique. PMID- 10680923 TI - Redefining the survival of the fittest: communication disorders in the 21st century. AB - OBJECTIVES: To determine the economic effect on the US economy of the cost of caring for people with communication disorders as well as the cost of lost or degraded employment opportunities for people with such disorders, including disorders of hearing, voice, speech, and language. STUDY DESIGN: Survey of available historical and contemporary governmental and scholarly data concerning work force distribution and the epidemiology of disorders of hearing, voice, speech, and language. METHOD: Analysis of epidemiological and economic data for industrialized countries, North America, and the United States. RESULTS: Communication disorders are estimated to have a prevalence of 5% to 10%. People with communication disorders may be more economically disadvantaged than those with less severe disabilities The data suggest that people with severe speech disabilities are more often found to be unemployed or in a lower economic class than people with hearing loss or other disabilities. Communication disorders may cost the United States from $154 billion to $186 billion per year, which is equal to 2.5% to 3% of the Gross National Product. CONCLUSIONS: Communication disorders reduce the economic output of the United States, whose economy has become dependent on communication-based employment. This trend will increase during the next century. The economic cost and the prevalence rates of communication disorders in the United States indicate that they will be a major public health challenge for the 21st century. PMID- 10680924 TI - Saline irrigation in the prevention of otorrhea after tympanostomy tube placement. AB - OBJECTIVES: Comparison of intraoperative saline irrigation to otic drops in the prevention of postoperative otorrhea in children with middle ear effusion undergoing bilateral myringotomy with ventilation tubes. STUDY DESIGN: This study was designed as a blinded, controlled, prospectively randomized trial. METHODS: Study children were randomly assigned to receive either otic drops for 3 days postoperatively or saline irrigation of the middle ear space at the time of myringotomy. Only children with effusion present at the time of surgery were included. All children were evaluated for drainage 7 to 14 days postoperatively, and the degree of drainage was graded from 0 to 4. RESULTS: Of the 84 patients entered into the study, 62 patients were eligible for data analysis (16 failed follow-up, 6 records were lost). Of the patients who completed the study, not all had bilateral effusions, resulting in 111 ears for inclusion in the study. Fifty two ears underwent irrigation, and 10 were noted to have otorrhea (19.2%). Fifty nine ears received otic drops, resulting in 21 ears with otorrhea (35.6%). Evaluating the degree of otorrhea with a five-point Leichert scale, the average score per ear was 0.42 for the saline irrigation group and 1.07 for the control group. The rate and degree of drainage were both statistically reduced in the saline irrigation group (P < .05). CONCLUSIONS: Using middle ear irrigation at the time of tympanostomy may be more effective than antibiotic drops in preventing postoperative otorrhea. PMID- 10680925 TI - Consequences to hearing during the conservative management of vestibular schwannomas. AB - OBJECTIVE: To estimate the risk of loss of serviceable hearing during the conservative management of vestibular schwannomas. STUDY DESIGN: Retrospective case review. METHODS: Twenty-five patients with a radiological diagnosis of unilateral vestibular schwannoma were managed conservatively for a mean duration of 43.8 months (range, 12-194 mo). The pure-tone average (PTA) (0.5, 1, 2, and 3 kHz) and speech discrimination scores (SDS) were measured at regular intervals throughout the entire duration of follow-up. Serviceable hearing was defined using two criteria: 70% SDS/30 dB PTA (the 70/30 rule) and 50% SDS/50 dB PTA (the 50/50 rule). The size and growth rate of tumors were determined according to the American Academy of Otolaryngology-Head and Neck Surgery guidelines (1995). Intervention was recommended if there was evidence of continuous or rapid radiological tumor growth, and/or increasing symptoms or signs suggestive of tumor growth. RESULTS: The risk of loss of serviceable hearing for the total group was 43% using the 70/30 rule and 42% using the 50/50 rule. Tumor growth was considered significant (> 1 mm) in 8 tumors (32%) and nonsignificant in 17 (68%). The risk of loss of serviceable hearing for the tumor-growth group was 67% using the 70/30 rule and 80% using the 50/50 rule. In contrast, the risk of loss of serviceable hearing for the no tumor-growth group was 25% using the 70/30 rule and 14% using the 50/50 rule. No audiological factors predictive of tumor growth were identified. CONCLUSIONS: There is a significant risk of loss of serviceable hearing during the conservative management of vestibular schwannomas. This risk appears to be greater in tumors that demonstrate significant growth. PMID- 10680926 TI - Dynamic asymmetry of the vestibulo-ocular reflex in unilateral peripheral vestibular and cochleovestibular loss. AB - OBJECTIVE: Rotatory tests in the horizontal plane have shown various degrees of vestibulo-ocular reflex (VOR) asymmetry in patients after surgical deafferentation of one labyrinth. The purpose of this work was to characterize dynamic horizontal VOR responses among patients presenting with a unilateral peripheral labyrinthine deficit of nonsurgical origin and to compare results in isolated vestibular loss versus cochleovestibular loss. STUDY DESIGN: This study included 40 patients who presented with an acute, spontaneous unilateral peripheral labyrinthine lesion. Twenty-two patients had vestibular loss alone (without associated hearing impairment) and 18 presented with a cochleovestibular deficit (sudden hearing loss with vertigo). The majority of these patients were part of a long-term protocol to evaluate vestibular compensation. METHODS: All patients underwent both the clockwise test and the counterclockwise rotatory test in the horizontal plane, using brief impulses of moderate intensity. Results were analyzed by a simplified model of vestibular function, allowing a parametric estimation of the response. RESULTS: A weak and transitory horizontal VOR asymmetry was observed in the 22 patients with vestibular loss. However, the 18 patients with cochleovestibular loss demonstrated a more severe and persistent asymmetry. CONCLUSIONS: This study revealed a difference in the dynamic characteristics of the horizontal VOR between patients with vestibular loss and those with cochleovestibular loss. Our results support the presence of an extensive labyrinthine lesion in cochleovestibular deficit that involves the otolith organs. The implications of this involvement on the central mechanisms of otolith-canal interaction are discussed. PMID- 10680927 TI - Pressure transfer between intracranial and cochlear fluids in patients with Meniere's disease. AB - OBJECTIVES: To elucidate the pressure transfer between intracranial and labyrinthine fluids in patients with well-defined unilateral Meniere's disease. STUDY DESIGN: Eleven patients previously exposed to hypobaric pressure agreed to be investigated further with the tympanic membrane displacement (TMD) technique. TMD was used to indirectly analyze perilymph pressure changes as the result of changes in body position. METHODS: Repeated measurements for both the diseased and the healthy ears were made with the patients supine and then in a sitting position. The TMD parameters for the maximum inward displacement, the Vi, and the mean volume displacement, the Vm, were calculated and compared. RESULTS: The paired comparison showed statistically significant larger Vi values for both ears in the supine position. A similar tendency was observed for the Vm value. This difference of the Vi was significantly larger for the diseased ear compared with the currently healthy ear. The results were compared to the audiometric and electrocochleographic results previously obtained on the same patients when they were subjected to hypobaric pressure. Patients who experienced the largest differences in hearing level thresholds in the lower frequencies also showed the greatest differences in TMD values as the result of postural changes. CONCLUSIONS: Despite the limited number, the statistically supported results suggest a relation between the efficiency of the routes of pressure transfer and the observed effect of hypobaric exposure. The results also indicate that for the patients tested, the routes of communication are more effective in the diseased ear than in the healthy ear--a condition that may relate to the pathogeneses of Meniere's disease. PMID- 10680928 TI - Temporal bone histopathology in connexin 26-related hearing loss. AB - OBJECTIVE: Mutations in GJB2, a gene that encodes a gap junction protein, Connexin 26 (Cx26), are responsible for approximately one third of sporadic severe-to-profound or profound congenital deafness and half of severe-to-profound or profound autosomal recessive nonsyndromic hearing loss (ARNSHL). Mouse mutants homozygous for knockouts of this gene are nonviable, precluding histopathologic studies of the associated inner ear pathology in this animal model. Therefore, we studied archival temporal bone sections to identify temporal bone donors with Cx26-related deafness. STUDY DESIGN: Temporal bone donors with a history of congenital severe-to-profound or profound deafness were identified in the registry of the Temporal Bone Library at the University of Iowa. Histological findings were interpreted in a blinded fashion. DNA extracted from two celloidin embedded mid-modiolar sections from each temporal bone was screened for the 35delG Cx26 mutation. The entire coding region of Cx26 was screened for other deafness-causing mutations if the 35delG mutation was detected. RESULTS: Of five temporal bone donors with congenital severe-to-profound deafness, one donor was found to have Cx26-related deafness. This individual was a Cx26 compound heterozygote, carrying the 35delG mutation and a noncomplementary Cx26 missense mutation on the opposing allele. Microscopic evaluation of this temporal bone showed no neural degeneration, a good population of spiral ganglion cells, near total degeneration of hair cells in the organ of Corti, a detached and rolled-up tectorial membrane, agenesis of the stria vascularis, and a large cyst in the scala media in the region of the stria vascularis. CONCLUSION: This study is the first to report the temporal bone histopathology associated with Cx26-related deafness. Preservation of neurons in the spiral ganglion suggests that long-term successful habilitation with cochlear implants may be possible in persons with severe-to-profound or profound Cx26-related deafness. PMID- 10680929 TI - Levels of intracellular protein and messenger RNA of mucin and lysozyme in normal human nasal and polyp epithelium. AB - OBJECTIVES: Mucus hypersecretion is a characteristic feature in chronic sinusitis with nasal polyps. The objective of this study is to examine whether the polyp epithelium itself contributes to a certain extent to the increased mucous secretions in chronic sinusitis with nasal polyps, and if it does, to determine which mucin genes are responsible for the increased mucin secretion. METHODS: Three pooled samples of normal nasal epithelial cells from each subject were obtained by scrapings from the inferior turbinates of 30 healthy adult volunteers and nasal polyps from 6 patients who underwent intranasal ethmoidectomy and polypectomy. Isolated epithelial cells were used for total RNA isolation for reverse transcriptase polymerase chain reaction and cell lysates for immunoblotting. RESULTS: The intracellular level of mucin from polyp epithelium was 2.9 times higher than that of normal nasal epithelium (P < .05). Interestingly, MUC2 and MUC8 messenger RNA (mRNA) levels were clearly upregulated in polyp epithelium compared with those of normal turbinate epithelium. CONCLUSIONS: Polyp epithelium can be considered to contribute in part to increased secretion in chronic sinusitis with polyps, and increased mucous secretion might be related to the increased mRNA level of MUC2 or MUC8 or both. PMID- 10680930 TI - Fontanelle and uncinate process in the lateral wall of the human nasal cavity. AB - OBJECTIVES: Although a complete anatomic knowledge of the fontanelle is a prerequisite to perform a surgical antrostomy opening, little is known about the boundary, shape, and size of the fontanelle. The purpose of this paper is to determine the best site for maintaining the patency of a surgical antrostomy opening by defining the anatomic boundaries, shape, and size of the fontanelle as well as its histological structure. MATERIALS AND METHODS: One hundred sagittally divided heads were utilized. Mucosa overlying the lateral nasal wall was carefully removed with an operating microscope under 6x magnification. In some cases, a double mucous membrane, including the posteroinferior portion of the uncinate process, was cut as a whole and embedded in paraffin. The sections were stained with H&E. RESULTS: The boundary of the fontanelle and the location of the natural ostium were described in detail. Eight patterns of the posteroinferior portion of the uncinate process were observed. There were three major fontanelle shapes when observed from the medial aspect to the lateral: triangular, pencil like, and oval. The triangular type was the most common. The anterior portion of the fontanelle was shorter than the posterior when observed medially and was wider than the posterior portion when observed inferiorly. CONCLUSIONS: The anterior portion of the fontanelle is more prone to stenosis than the posterior portion. An antrostomy in the posterior fontanelle may be more ideal for a middle meatal antrostomy of the maxillary sinus. PMID- 10680931 TI - Extradural extranasal combined transmaxillary transsphenoidal approach to the cavernous sinus: a minimally invasive microsurgical model. AB - The authors have previously described an extradural transmaxillary approach to the anterior compartment of the cavernous sinus. In an effort to expand the surgical access to that area without necessitating a craniotomy or wide transfacial dissection, they present a modification of the transmaxillary approach to the sellar region and cavernous sinus. METHODS: The approach was developed on 12 fresh and 12 embalmed cadaveric specimen, and 2 dry skulls. The initial sublabial incision is followed by a maxillotomy to expose the course of the infraorbital nerve (terminal branch of maxillary branch of the trigeminal nerve) on the roof of the maxillary sinus. The route of the infraorbital nerve is traced to the pterygopalatine fossa as a guide to the foramen rotundum. Superomedial drilling of the foramen rotundum is then performed to reveal the contents of the superior orbital fissure. After the nerves are safely identified in the superior orbital fissure, medial enlargement of the window into the cavernous sinus is made possible by drilling the lateral and posterior wall and septum of the sphenoid sinus. RESULTS: The combined transmaxillary transsphenoidal approach offers an excellent exposure of the sellar and infrasellar region. The approach offers clear visualization of the ipsilateral loop of the carotid artery, the pituitary fossa, and the cranial nerves of the ipsilateral cavernous sinus. Mean operative reach is 38 mm from the posterior wall of the maxillary sinus to the ipsilateral carotid loop and 56 mm to the contralateral loop. The width of the operative window is 26 mm at the base within the cavernous sinus. CONCLUSION: The model offers a minimally invasive approach that avoids the need for craniotomy or violating the nasal cavity. It may be safely employed to access vascular as well as invasive lesions of the sellar and infrasellar region. The approach offers excellent visualization of the ipsilateral intracavernous carotid artery with both proximal and distal control, as well as cranial nerves III, IV, VI, V2, the hypophyseal region, and the medial aspect of the contralateral cavernous sinus. PMID- 10680932 TI - Endoscopic sinus surgery using intraoperative computed tomography imaging for updating a three-dimensional navigation system. AB - OBJECTIVES: The use of three-dimensional navigation systems provides information on the structures surrounding the field of operation and thereby reduces the risk of iatrogenic damage. The computed tomography (CT) data conventionally used are provided by preoperative scanning procedures, which means that tissue changes coming about during surgery are not seen on the screen. An intraoperative CT scanning procedure being able to update the CT data could provide a solution. STUDY DESIGN: Endoscopic sinus operations using an intraoperative CT updating the three-dimensional navigation system were performed on six persons to find out, whether the above is true. METHODS: Different parameters, advantages, and disadvantages in the cases of these six patients were compared with a group of 22 patients who underwent conventional endoscopic sinus surgery with different three dimensional navigation systems without updating the CT data set. RESULTS: The intraoperative CT for updating the three-dimensional navigation system provides useful information for the surgeon. CONCLUSION: Balancing its advantages against its disadvantages, the updating of the CT data set with intraoperative CT cannot be recommended for conventional standard endoscopic sinus surgery. PMID- 10680933 TI - Role of vascular reflex in nasal mucosal swelling in nasal allergy. AB - OBJECTIVE: In patients with nasal allergy, antigen challenge on the unilateral nasal mucosa results in nasal secretion not only in the ipsilateral but also in the contralateral nasal cavities that can be inhibited almost completely by premedication with atropine sulfate. The present study was performed to elucidate if centrally mediated vascular reflex induced by antigen challenge plays a role in nasal mucosal swelling in subjects with nasal allergy. METHODS: Variations of mucosal swelling and mucosal blood flow in the ipsilateral and the contralateral nasal cavities after unilateral antigen challenge were evaluated by acoustic rhinometry and laser Doppler flowmetry in 20 patients with perennial nasal allergy. RESULTS: Unilateral antigen challenge caused ipsilateral and contralateral nasal mucosal swelling in 17 and 13 patients, respectively. Incidence of contralateral nasal mucosal swelling after unilateral antigen challenge was significantly higher compared with that after control disc challenge (P < .001). In 10 patients in whom unilateral antigen challenge caused bilateral nasal mucosal swelling, significant swelling of the nasal mucosa lasted for more than 30 minutes in the ipsilateral nasal cavity after antigen challenge compared with only 15 minutes in the contralateral nasal cavity. Peak values of contralateral mucosal swelling were 45.3% of those of ipsilateral nasal mucosa. CONCLUSIONS: Centrally mediated vascular reflex is partially involved in the onset of nasal mucosal swelling observed after antigen challenge in subjects with nasal allergy. However, nasal mucosal swelling that persists and proceeds even 20 minutes after antigen challenge is caused by the direct effects of chemical mediators on the nasal vasculature. PMID- 10680934 TI - Assessing olfactory function in laryngectomees using the Sniffin'Sticks test battery and chemosensory evoked potentials. AB - OBJECTIVES/HYPOTHESIS: Laryngectomees are often considered to be completely anosmic. The aim of this study was to determine whether anosmia in laryngectomees reflects diminished transport of odorants to the olfactory epithelium or olfactory epithelial damage. STUDY DESIGN: Twenty-five laryngectomees were examined psychophysically using the Sniffin'Sticks test battery. All patients rated the degree of their subjectively perceived deficit on a rating scale. Chemosensory evoked potentials were also recorded in 11 of 25 patients. RESULTS: Sixteen patients complained very little about their smell deficit, even though the psychophysical testing found 18 patients to be anosmic and 7 hyposmic. Olfactory potentials could be recorded in only 7 of the 11 patients who received this evaluation, despite the fact that all 11 perceived, at least vaguely, the olfactory stimulus, H2S, during the recording sessions. No meaningful correlation between the psychophysical and electrophysiological data was observed. CONCLUSIONS: The psychophysical data revealed the laryngectomees to be either functionally anosmic or hyposmic. The olfactory evoked potential data suggested that at least in two thirds of the laryngectomized patients the olfactory system had some function, even up to 22 years after surgery. Because patients in everyday situations find ways to bring odorants to the olfactory mucosa, the low number of complaints about subjectively perceived deficits is reasonable. PMID- 10680935 TI - Arterial supply of the nasal tip in Asians. AB - OBJECTIVES: The blood supply to the nasal tip and columella was examined to determine whether it could be damaged as a result of transcolumellar incision during an external rhinoplasty approach in Asians. METHODS: The blood vessels that supply the nasal tip were examined by dissecting 51 cadavers, and their corresponding 102 nasal sections were injected with red latex before dissection. The size and distribution of the vessels were measured with the unaided eye and the primary supply vessels were determined. The subdermal layer in which the vessels lie and the course of the vessels were also investigated. RESULTS: The main blood supply source of the nasal tip proved to be the lateral nasal artery in 78% (80/102) of the cases examined, while the remaining cases (22%) received their blood supply via the dorsal nasal artery. Columellar branches were narrow in diameter and varied in size and appearance, and were therefore appeared insufficient as a main blood supply. These arteries passed through the musculoaponeurotic layer, but they were also in close proximity to the main surgical plane in the dome of the lower lateral cartilage. CONCLUSIONS: The authors speculate that the nasal tip blood supply in Asians is primarily derived from the lateral nasal or dorsal nasal arteries, with a variable contribution from the columella arteries. Therefore, it is important to correctly determine the surgical plane below the musculoaponeurotic layer in order to prevent skin flap necrosis or nasal tip deformity that may occur from damage to the main vessel during an external rhinoplasty approach. PMID- 10680937 TI - Interventional sialendoscopy. PMID- 10680936 TI - Long-term use of preservatives on rat nasal respiratory mucosa: effects of benzalkonium chloride and potassium sorbate. AB - OBJECTIVES: The preservatives benzalkonium chloride (BZC) and potassium sorbate (PS) are widely used, not only for nasal drops, but also for eyedrops and cosmetics. However, there have been many case reports that consider lesions such as dermatitis or conjunctivitis to be the results of irritation induced by BZC or PS. METHODS: We evaluated the histological changes after the long-term administration of BZC or PS on rat nasal respiratory mucosa. Forty rats were used for the BZC group and 40 rats for PS group. Animals in each group were divided into four subgroups The first subgroup received a low-concentration preservative solution that was commonly used for nasal sprays. The second subgroup received a high-concentration preservative solution that was reported to induce dermatitis in humans. The third and fourth subgroups received a steroid mixed preservative solution of low and high concentrations, respectively. The control group was administrated normal saline. After each group received 1, 2, and 4 weeks of topical administration, the symptomatic and histological changes on H&E stain were observed. RESULTS: Sneezing and nasal rubbing with forelegs were observed in almost all subgroups by the seventh day of treatment. The preservatives induced nasal lesions, including intraepithelial glandular formation, inflammatory cell infiltration, vascular hyperplasia, and edematous change. The symptomatic and histological changes were pronounced with the prolonged duration of administration. Similar results were observed in the steroid mixed-solution groups. In the PS steroid mixed-solution group, however, symptoms and nasal lesions were reduced with the prolonged duration of administration. CONCLUSION: It is our finding that even a low-concentration solution of preservative can lead to nasal lesion. Hence there is a strong need to develop both a preservative that can be safely and widely used and a nasal spray without preservatives. PMID- 10680938 TI - Antimicrobial activity of synthetic salivary peptides against voice prosthetic microorganisms. AB - OBJECTIVES: To investigate whether synthetic salivary antimicrobial peptides have an inhibitory effect on the growth of bacteria and yeasts isolated from used silicone rubber voice prostheses. METHODS: The antimicrobial activities of six synthetic salivary peptides (histatin 5, dhvarl, dhvar4, dhvar5, lactoferrin b 1730 [LFb 17-30], and cystatin S1-15) at concentrations of 2 and 4 mg/mL were determined against different oropharyngeal yeast (four) and bacterial (eight) strains and against a "total microflora" isolated from explanted voice prostheses using agar diffusion tests. The spectrum of susceptible microorganisms was determined qualitatively. RESULTS: Histatin 5 and cystatin S1-15 did not show any antimicrobial activity against the microorganisms involved in this study. Dhvar1 was active against some of the oropharyngeal microorganisms tested, including the yeast strains, but not against Rothia dentocariosa, Staphylococcus aureus, Escherichia coli, and the total microflora Dhvar4 was active against all microorganisms tested, including the total microflora. Dhvar5 lacked activity against E coli and the total microflora LFb 1730 did not inhibit the growth of any of the yeast strains involved and showed only minor activity against some of the bacterial strains. LFb 1730 slightly inhibited the growth of the total microflora from an explanted prosthesis. CONCLUSIONS: The synthetic salivary peptide dhvar4 has a broad antimicrobial activity against all microorganisms that are commonly isolated from explanted voice prostheses, including yeasts. Therewith, it may represent a useful drug, as an alternative for antibiotics and antimycotics employed in various ways to prolong the lifetime of voice prostheses in laryngectomees. PMID- 10680939 TI - Postoperative airway findings after maxillomandibular advancement for obstructive sleep apnea syndrome. AB - OBJECTIVE: To evaluate the upper airway characteristics in the early postoperative period after maxilomandibular advancement for obstructive sleep apnea syndrome. METHODS: Nasopharyngolaryngoscopy was performed before and 48 hours after surgery on 70 consecutive patients who underwent maxillomandibular advancement for obstructive sleep apnea syndrome. The preoperative and the postoperative evaluations were performed by the same examiner for consistency. RESULTS: Mild to moderate lateral pharyngeal wall edema was identified in 70 consecutive patients. Fourteen patients (20%) had edema as well as ecchymosis involving the pyriform sinus and aryepiglottic fold. Four of these patients (6%) were also noted to have hypopharyngeal hematoma involving the pyriform sinus, aryepiglottic fold, arytenoid, and false vocal cord that partially obstructed the airway. These four patients were closely monitored for 1 to 2 additional days for possible expanding hematoma leading to airway compromise. None of these patients were found to have airway difficulty, and the minimum oxygen saturation was more than 90% throughout the hospitalization. All four patients were discharged uneventfully, and the hematoma resolved completely within 10 days. CONCLUSION: Although postoperative edema was expected after maxillomandibular advancement, hypopharyngeal hematoma was unexpected. Although none of our patients had evidence of airway difficulty, the possibility of an expanding hypopharyngeal hematoma should be considered in patients complaining of breathing difficulty after maxillomandibular advancement surgery. PMID- 10680940 TI - Predictive factors for complications in children with laryngeal damage at extubation. AB - OBJECTIVES: To determine which factors contribute to early complications when intubated children show macroscopic lesions at extubation. STUDY DESIGN: Retrospective review of 96 consecutive medical records of children aged 1 day to 15 years. Patients were divided into three groups depending on the extent of the subsequent treatment required: medical, reintubation, and surgical. METHODS: Age, sex, clinical history, and macroscopic features of the lesions were collected and data were compared in each group. RESULTS: Underlying noninfectious respiratory diseases and young age were found to be risk factors for higher incidence of complications, but not prolonged or multiple intubations. Edema, especially in the glottic area, was a risk factor for surgical treatment. Multiple lesions were risk factors for reintubation. CONCLUSIONS: History of intubation, its cause, and lesions discovered at extubation can provide the basis for definition of an "at risk" profile for intubated children. PMID- 10680941 TI - Endonasal laser surgery with a new laser fiber guidance instrument. PMID- 10680942 TI - New millennium, new values: citizen participation as the democratic ideal in health care. PMID- 10680943 TI - Prenatal care in the first trimester: misleading findings from HEDIS. Health Plan Employer Data and Information Set. AB - OBJECTIVE: To understand factors influencing Health Plan Employer Data and Information Set (HEDIS) rates for the measure 'Prenatal care in the first trimester'. DESIGN: Telephone survey of a retrospective cohort of women with a live birth. Medical record review of a sample of both responders and non responders to the telephone survey. Detailed review of HEDIS data collection procedures. SETTING: A managed care plan in California. STUDY PARTICIPANTS: Women aged 18-49 years at date of delivery, who delivered a live birth from 1 October 1995 through 31 March 1996, and who were continuously enrolled in a California managed care plan for 12 months prior to delivery (telephone survey, n= 1,185; medical record review, n= 465). RESULTS: Of the women participating in the telephone survey, 95% indicated that their first prenatal visit occurred during the first 3 months of pregnancy. Using HEDIS 3.0 standards, a review of medical records for a sample of these women indicated that 94% of the women initiated care during the first trimester. These results contrasted sharply with 1995 and 1996 HEDIS rates of 64% and 75%, respectively. CONCLUSION: An investigation of the discrepancy between HEDIS rates and rates from both telephone survey and medical record review led to the finding that the low HEDIS rates were due not to a true low rate of early care, but to data collection problems, including difficulty obtaining medical records. Potential solutions involving health plan activities, revisions to the official HEDIS process and revised reporting of results are proposed. PMID- 10680944 TI - How do we compare with our colleagues? Quality of general practitioner performance in consultations for non-acute abdominal complaints. AB - OBJECTIVE: To investigate what factors influence the quality of general practitioner performance in consultations for non-acute abdominal complaints and to establish the extent to which performance quality differs between general practitioners (GPs). DESIGN: Explorative study in two parts: (i) detection of variables influencing quality scores of consultations; and (ii) comparison of mean quality scores of the consultations, selected by each GP. SETTING: Sixty-two family practices across The Netherlands. SUBJECTS: Eight-hundred and forty consultations concerning non-acute abdominal complaints, first encounters; 62 GPs. METHOD: Multilevel analysis was carried out to detect factors that influence quality. After correction for the effect of significant factors the mean quality scores of individual GPs were calculated and compared. RESULTS: Eighty-eight per cent of the total variance in quality scores was located at the consultation/patient level, and 12% at the GP level. One consultation characteristic had significant influence on quality: quality scores were higher in consultations of longer than average duration (>15 minutes). Several patient characteristics were of significant influence. Consultation quality scores were higher in consultations for patients with upper abdominal or non-specific abdominal complaints. Quality scores were lower in consultations with female patients and with patients aged >40 years. Together these characteristics explained 20% of the variance at the GP level. None of the GP characteristics investigated in this study appeared to have significant influence on the quality of their performance. After correction of the scores for the effect of significant factors the differences in performance quality between GPs remained significant. CONCLUSIONS: Quality of performance is far more influenced by consultation and patient characteristics than by GP characteristics. After correction for influencing factors, the mean quality scores of GPs still differed considerably and significantly. For many GPs the quality scores varied substantially between different consultations; to a large extent this variation remained unexplained. Consultation quality can be improved by booking more time per patient and by giving more medical/technical attention to female and older patients. PMID- 10680945 TI - Physician attitudes, self-estimated performance and actual compliance with locally peer-defined quality evaluation criteria. AB - BACKGROUND: Physicians' agreement with quality evaluation criteria, and estimates of their own and their colleagues' compliance with these criteria were compared with actual compliance. METHODS: Physicians practicing in 10 health centers in Spain defined 13 quality evaluation criteria for two patient conditions (upper respiratory infections and high serum cholesterol). Compliance with criteria was measured by an external team, using random samples of medical records stratified by condition in each health center (n= 1,000). Concurrently, physicians were surveyed regarding agreement with the criteria, and were asked to estimate their own and their health center's rate of compliance with these criteria. RESULTS: Agreement ratings varied from 5.9 to 9.1 on a 10-point scale. Actual compliance rates ranged from 1.8 to 91.7% of records. Agreement correlated significantly with self-reported compliance but not with actual compliance. Estimates of one's own and one's health center compliance were positive and significantly correlated for all criteria, but were significantly higher for oneself than for one's health center for six of 13 criteria. CONCLUSIONS: Wide variation in physicians' agreement on quality criteria and in actual performance reveal a lack of clear guidelines. Agreement on criteria did not always translate into compliance with criteria. Physicians tended to rate their own performance as better than the average of their peers, suggesting that aggregate data may not influence physicians to change. Self-estimate of one's own or one's colleagues performance is not a good proxy for actual performance so that peer ratings are of dubious value for performance appraisal. PMID- 10680946 TI - Using patient surveys to measure the quality of outpatient care in Krakow, Poland. AB - OBJECTIVE: To test the feasibility of using patient reported information to create indicators of quality (access, patient experience--including satisfaction, and clinical quality) with the goal of providing Krakow city clinic managers (and potentially other audiences) with information about the quality of outpatient care in selected clinics. Setting and methods. Almost 2,000 patients from 19 outpatient clinics in Krakow, Poland were surveyed in November and December 1997 and January 1998. We prepared a self-completed questionnaire to capture data about the patient's experience with access to services, interactions with registration staff, communication with the doctor, information received from the doctor, and receipt of preventive services. RESULTS: Access varied across clinics. For example, 84% of patients waited less than 10 minutes at registration, whereas only 53% of patients waited less than 30 minutes to see the doctor. Among those who tried to register by telephone, only 72% were successful. Satisfaction was highest with the doctor visit (satisfaction=79, on a scale of 1 100) and lowest with telephone registration (satisfaction = 59). Preventive health care screening was generally disappointing, particularly for Papanicolaou smear and clinical breast examination, although frequent users of a clinic (with more opportunities for screening) generally had higher rates of screening. CONCLUSION: We demonstrated the feasibility of constructing indicators of multiple dimensions of the quality of outpatient care using patient-reported information. Quality dimensions captured by survey included access, patient experience and clinical quality. Results were successfully summarized in easy to read and understand formats for clinic managers and city health department officials. PMID- 10680947 TI - Construction of a scale measuring inpatients' opinion on quality of care. AB - OBJECTIVE: To develop a reliable and valid measure of patient opinions on quality of hospital care. DESIGN: Issues of importance to patients and possible scale items were generated by literature review and non-structured interviews of patients, former patients, health care providers and researchers. Semi-structured interviews with inpatients and pilot studies were conducted to modify or remove ambiguous questions and reduce skewed responses. A study was then made to select from these questions relevant items and variables correlated to patient evaluation of quality of care. A principal-components analysis was performed to select items and assess construct validity. Cronbach's alpha coefficients were calculated to estimate the reliability of the scale. Time reliability and concurrent validity were also considered. SETTING: An 800-bed French short-stay teaching hospital in Paris. STUDY PARTICIPANTS: Five-hundred and thirty-four consecutive patients hospitalized in eight medical and surgical wards. RESULTS: A 26-item scale was developed. Component analysis indicated two subscales: 'medical information' and 'relationship with staff and daily routine'. Levels of reliability were satisfactory: Cronbach's alpha coefficient exceeded 0.87 for overall scale and subscales. Concurrent validity and time reliability were also satisfactory. Multivariate analysis showed that, taking into account patients and hospitalization characteristics linked to scores (age, health status, number of hospitalizations, comorbidity, time since diagnosis, admission pattern, private patient and difficulties reported by staff), these scores differed among departments. CONCLUSION: A reliable, valid measure of inpatients' opinions on quality of care has been developed in a French hospital and variables that have to be taken into account to compare hospital departments have been selected. Items selected in the scale emphasized the importance that patients give to receiving medical information. PMID- 10680948 TI - Quality of diabetes care in a university health center in Lebanon. AB - OBJECTIVE: To assess the quality of care provided to diabetic patients by family physicians in a university health clinic, using measures of glycemic and cardiovascular risk control as well as documentation of and adherence to World Health Organization (WHO) guidelines for diabetes primary care. DESIGN: Chart review of the previous year's medical notes for all identified diabetics in the practice over 2.5 years. RESULTS: Two-hundred and four diabetic patients were identified, with an estimated prevalence of 4.1%. The majority was type II diabetics, on oral hypoglycemic agents. Glycosylated hemoglobin was documented in 39.7% of patients, fasting plasma glucose in 99%, cholesterol in 93.1%, triglycerides in 91.2% and blood pressure in 85.8%; optimal control of these indicators was noted in 28.4%, 17.8%, 34%, 29.6% and 55.4% respectively. Fifty percent of the diabetics were referred for retinal checks. Physicians documented the presence of nephropathy in 46.8% and neuropathy in 59.6%; however, they documented patient instruction on foot care, diet, exercise and diabetes self care poorly. CONCLUSION: There is a need for interventions to improve management and documentation in diabetes care in order to achieve early detection and prevention of complications. Developing a protocol for the clinic based on standard guidelines, and the use of flow sheets may be helpful in improving these intermediate indicators of quality of care. PMID- 10680949 TI - Determinants of users' satisfaction with primary health care settings and services in Saudi Arabia. AB - OBJECTIVE: To identify the components of primary health care that cause most concern to service users and to identify socio-demographic and other factors associated with satisfaction among the users of primary health care centres. DESIGN: Interviews conducted by well-trained interviewers with a random sample of heads of households. The questionnaires were composed of questions that measure the extent of satisfaction with settings and services in the primary health care centres using a 5-point rating scale from very satisfied to very dissatisfied. SETTING: The community of Qateef, eastern Saudi Arabia. STUDY PARTICIPANTS: A sample of 802 households representing 838 families was chosen randomly from the housing lists of the primary health care centres in Qateef. There were 40 vacant houses and nine refusals. Thus the number of heads of households actually interviewed was 789. RESULTS: Waiting area structure, confidentiality measures and environmental structure were the areas that caused most concern to service users. The factors that showed the greatest association with satisfaction were the type of the primary health care centre building (purpose-built or rented), literacy status of the household head (literate or illiterate), the extent of the primary health care centre utilization (regular or infrequent). Surprisingly, age showed no association when other characteristics of the respondents were adjusted for, and sex was less important than in other studies. CONCLUSION: How regular the respondent was in using his or her primary health care centre was more predictive in deciding the extent of satisfaction with the various components in the study than the other variables. Socio-demographic factors played minor roles in deciding the extent of satisfaction, although each had a deciding role with one or more, but not all, components. PMID- 10680950 TI - Communication gaps in paediatric care. PMID- 10680951 TI - Chicken antibodies: a clinical chemistry perspective. AB - The chicken immune system has been studied for many years and these studies have contributed substantially to our understanding of the fundamental concepts of immunology and the development of different immunoglobulin classes. It is thus surprising that only a small fraction of the antibodies presently used in laboratories are of avian origin. A laying hen produces more yolk antibodies than a rabbit can produce during the same time period, and the animal care costs are lower for the chicken compared to the rabbit. Chicken antibodies offer many advantages to the traditional mammalian antibodies when used for the detection of mammalian antigen. Due to the evolutionary difference chicken IgY will react with more epitopes on a mammalian antigen, which will give an amplification of the signal. Chicken antibodies can also be used to avoid interference in immunological assays caused by the human complement system, rheumatoid factors, human anti-mouse IgG antibodies (HAMA) or human and bacterial Fc-receptors. The antibodies can be purified in large amounts from egg yolk, making laying hens highly efficient producers of polyclonal antibodies. PMID- 10680952 TI - Exercise-induced bronchoconstriction in adults with asthma--comparison between running and cycling and between cycling at different air conditions. AB - The bronchial response to cycling and running was compared in six adult asthmatic persons. The effects of different air conditions during cycling regarding the induction of bronchoconstriction was studied. The exercise consisted of 6 minutes' work at an intensity of 80-85% of maximal heart rate. Heart rate, oxygen consumption and ventilation were measured to check that the exercise level was the same in all tests. Peak expiratory flow (PEF) was used to test for bronchoconstriction. Bicycling and treadmill running were performed under indoor conditions and bicycling while breathing cold, dry air (-18 degrees C) and room tempered humid air (60% RH), respectively. No difference in bronchoconstriction was found between cycling and running under indoor conditions. However, bicycling exercise with inhalation of cold dry air provoked more bronchoconstriction than when inhalating humid air (PEF reductions of 19.4+/-6% and 6.1+/-2%, respectively). No differences were found between the exercise modes in heart rate, oxygen consumption, ventilation per minute, respiratory rate, carbon dioxide elimination or subjective ratings of perceived exertion and breathlessness. It is concluded that it is not the type of exercise, but the ventilation demand and humidity of the inspired air that are the main determinants of the occurrence and degree of bronchoconstriction. PMID- 10680953 TI - Expression of prostasome-like granules by the prostate cancer cell lines PC3, Du145 and LnCaP grown in monolayer. AB - Prostasomes are a granular type of secretory product in the human prostate gland cells. It is not known, whether in vitro grown cells derived from human prostate cancers also express prostate secretory components containing granules with properties similar to the prostasomes. Therefore, we carried out the present investigation and found that cytospins of in vitro grown PC3, DU145 and LNCaP cells generally expressed a granular secretion. DU145 demonstrated the highest ratio of cells with granules (about 90%), while cytospins of PC3 and LNCaP contained less stained cells (50-70%). Purified granules from PC3 cells were immunoreactive with a monoclonal antibody (mAb78) originally raised against human seminal prostasomes. The PC3 granules also shared the property with human seminal prostasomes having an elevated UV260/UV280 absorbance ratio. On the other hand we found a low aminopeptidase activity in PC3 granules contrary to that of human prostasomes. Prostasomes may form a heterogeneous group with different properties due to the source from which they are isolated and perhaps it is justified to recognize them as different members of a prostasome family. PMID- 10680954 TI - Poorly differentiated, solid-type adenocarcinoma of the stomach. AB - Data of 58 cases of poorly differentiated, solid-type adenocarcinoma of the stomach treated at our hospital between 1985 and 1995 were reviewed and compared to data of 146 cases of non-solid-type carcinoma in order to determine whether there are distinguishable clinicopathological features between these two types of carcinoma. Significant differences were observed with respect to tumor size, stage, macroscopic appearance, depth of invasion, histologic growth pattern, lymph node metastasis, microscopical lymphatic invasion and vascular permeation. Patients in the solid-type cancer group tended to have smaller tumors; the disease was in the early stage in 48% of the patients, and total gastrectomy was performed in only 20 of the 58 patients. Nodal involvement, lymphatic invasion and vascular permeation were also less common in patients with solid-type cancer. The overall survival rate of patients with solid-type carcinoma was higher than that of patients with non-solid-type carcinoma, though no significant differences were observed when corrected for stage. Our results suggest that poorly differentiated solid-type carcinoma of the stomach should be regarded as a distinct type of adenocarcinoma that has a good prognosis. The significant prognostic factors for this type of gastric cancer are lymphatic invasion and tumor location. PMID- 10680955 TI - Clinicopathologic prognostic features in patients with gastric cancer associated with esophageal or duodenal invasion. AB - BACKGROUND: We evaluated the influence of several clinicopathologic variables on 5-year survival of patients with gastric cancer associated with esophageal or duodenal invasion, and determined the significance of resection line involvement. PATIENTS AND METHODS: A review of the database for gastric adenocarcinoma at Sendai National Hospital between January 1985 and December 1995 identified 923 patients who underwent gastric cancer resection. Of these patients, 37 were reported to have tumour infiltration of the esophagus or duodenum on histological examination of the resected specimens. Univariate and multivariate analyses of patients with esophageal or duodenal invasion were performed to evaluate the prognostic significance of clinicopathologic features. Then the patients were divided into two groups based on the results of microscopic examination: a tumour wedge-positive group for resection margins of less than 5 mm in width and a tumour wedge-negative group for resection margins of more than 5 mm in width. There were 8 patients in the narrow (margin-positive) group and 29 patients in the wide margin (margin-negative) group, respectively. RESULTS: Univariate analysis revealed that the significant prognostic factors were nodal involvement (p=0.0004) and gross type (p=0.0031). Multivariate analysis of the esophagus or duodenum-invaded cancer cases, however, revealed that only nodal involvement was a significant prognostic factor. There were statistical correlations between these groups (margin-positive and margin-negative groups) and the Borrmann type of tumour and tumour size. The survival rate was worse in patients with tumour line involvement. CONCLUSIONS: Multivariate analysis revealed that the prognosis of patients with esophageal or duodenal invasion was affected only by nodal involvement independently. The risk of surgical margin involvement was high in cases of a large Borrmann type-4 tumour and infiltrative carcinoma. PMID- 10680956 TI - Malignant fibrous histiocytoma: a method to control intraoperative hemorrhage by clamping the feeding arteries. AB - Malignant fibrous histiocytoma (MFH), also referred to in the past as malignant fibrous xanthoma and fibroxanthosarcoma, is a tumor of mesenchymal tissue origin. A case of retroperitoneal MFH was reported. In this paper, we describe a method of hemostasis and intraoperative control of hemorrhage during resection of retroperitoneal MFH by snaring the feeding arteries. The patient was successfully operated on using this technique. PMID- 10680957 TI - A qualitative study on changes in local brain pH due to discrete cerebral microembolism. AB - In this work autoradiography of 14C-5,5-dimethyl-2,4-oxazolidinedione (14C-DMO) was used to trace changes in local cerebral pH in embolized awake rabbits. One hour after i.v. injection of 14C-DMO small cerebral ischemic foci were produced in rabbits by injecting plastic beads into the left heart ventricle under short acting anaesthesia, and after another hour the animals were put to death and their brains processed for autoradiography of 14C-DMO. Evidence of acidosis was in general not found in the microischemic regions, though there were a few possible exceptions. However in the hippocampus a diffuse acidosis involving a large part of the structure, could be found in 2 of the 4 experiments. This hippocampal phenomenon probably reflected the same process as has been observed using autoradiography of 2-deoxyglucose (reflecting cellular glucose uptake) on the same ischemic model increased 2-deoxyglucose phosphorylation. Because the hippocampus is involved in the memory function and the fact that small infarcts are coupled to dementia, this phenomenon should be drawn into focus for further studies. PMID- 10680958 TI - A mathematical model of in situ freezing in liquid nitrogen. AB - In situ freezing is a procedure, typically applied in neuroscience, to halt metabolism and diffusion. However, the freezing process is not instantaneous, and the regional concentrations of a compound under study may change before the tissue is completely frozen. Knowing the local freezing time, metabolic rate and the diffusion coefficient of the compound of interest, it should be possible to reconstruct the spatial concentration profile prevailing before the object was placed in the cryogen. A mathematical model for calculating the temperature changes at different depths in rabbit and rat heads cooled in liquid nitrogen has been developed. By comparing with experimental results it has been found that the mathematical model can be used for prediction of the local freezing time with a small error. PMID- 10680959 TI - Evaluation of the teratogenic potential of pyrazinamide in Wistar rats. AB - We have tested Pyrazinamide (PZA), an essential component of modern short-course tuberculosis treatment regimen, for teratogenicity using Wistar rats. The drug was given by oral intubation from 6-15 days of gestation, at doses of 0, 25, 100 and 500 mg/kg body weight per day. Reduction in body weight and food consumption were observed in the treated dams. On day 20 of gestation, all the dams were killed by cervical dislocation and signs of maternal toxicity, reproductive indices and fetal measurements were recorded. Dams given doses of 100 and 500 mg/kg had significantly higher incidence of reabsorbed fetuses, reduced litter size, and impaired neonatal growth than those given no PZA or only 25 mg/kg dose. External visceral and skeletal examination of all fetuses of PZA-treated dams showed several types of variations which were neither dose related nor having a consistent pattern. However, these variations occurred mostly in the dams treated with the dose of 500 mg/kg. In conclusion, these data show that in Wistar rats, only high doses of PZA (100 and 500 mg/kg) produced fetotoxicity. No evidence of teratogenic effect of the drug was observed. PMID- 10680960 TI - Assessment of template quality by the incorporation of an internal control into a RT-PCR for the detection of rabies and rabies-related viruses. AB - A method is described to assess RNA template quality by the incorporation of a ribosomal RNA (rRNA) internal (in tube) control into a standard rabies and rabies related virus specific RT-PCR. Specific virus and rRNA templates were co amplified in a duplex reaction from RNA extracts derived from 60 isolates representing all six of the established lyssavirus genotypes. To ensure a wide species applicability of this technique we demonstrated that the rRNA assay was capable of functioning using the cells or tissues of 14 different mammals. Parallel studies between the duplex and the unlinked lyssavirus assay demonstrated only a minor reduction in the sensitivity of the former test. The ribosomal and viral targets (unlike beta-actin RNA) were shown to have similar degradation kinetics making rRNA amplification a good control for viral target integrity. As a consequence, the use of this system would reduce the likelihood of obtaining false negative RT-PCR results from lyssavirus infected material. PMID- 10680961 TI - A simple purification and fluorescent assay method of the poliovirus 3C protease searching for specific inhibitors. AB - Picornaviruses such as poliovirus, foot-and-mouth disease virus, and encephalomyocarditis virus produce their proteins by translating their genomic RNA, injected within the host cell, into a precursor polyprotein, which is then subjected to precise processing. The polyprotein is cleaved into mature proteins predominantly by the viral 3C protease. A simple purification and assay method for poliovirus 3C protease for use for screening for inhibitors of the 3C protease is described. A poliovirus cDNA fragment containing the 3C protease coding region was inserted into pET22b vector and expressed in Escherichia coli. The His-tagged protein (3CD'-His) was purified by a Ni-affinity column and the activity of the purified enzyme was measured by a fluorescent assay with a fluorogenic substrate containing the 3C-specific cleavage site, MocAc-MEALFQGPLQY Dnp. The kinetic parameters calculated from the Lineweaver-Burk plot and the effects of inhibitors showed that E. coli expression with His tag and the assay using the fluorogenic substrate are efficient, simple and sensitive methods for purifying the 3C protease, and measuring its activity. PMID- 10680962 TI - Green fluorescence protein as a transcriptional reporter for the long terminal repeats of the human immunodeficiency virus type 1. AB - Using the enhanced green fluorescence protein (EGFP), a transient reporter expression system was established to assess the transcriptional activity of the long terminal repeats (LTR) of primary isolates of the human immunodeficiency virus type 1 (HIV-1). Consistent with the conventional chloramphenicol acetyl transferase (CAT) reporter, EGFP expression, under the direction of HIV-1 LTR, was readily detected in the transient transfection and was elevated by co transfection of HIV-1 tat-expression vector. Comparing to CAT, however, EGFP expression system has two advantages: (i) Using a fluorescence activated cell sorter (FACS), it was possible to simultaneously measure transfection efficiency and fluorescence intensity of the transfected live cells without the necessity of co-transfection of a reference plasmid for comparing the transcriptional activity of two promoters; and (ii) EGFP expression was readily detected at a DNA concentration where CAT activity was not detectable possibly because the transfectants could be 'gated'. On the other hand, at a higher concentration of DNA, CAT signal became more prominent than that of EGFP, possibly because the enzymatic activity of CAT 'amplified' the signal. EGFP fluorescence detected by FACS was a direct measurement of the expressed chromophore. It is concluded that the system is rapid, reproducible, convenient and useful for quantitative analysis of transcription. PMID- 10680963 TI - The effects of cell passages on the cell morphology and the outcome of herpes simplex virus type 1 infection. AB - Because cell cultures are essential in biological research which involves the analysis of virus morphogenesis, this study focused on examining the significance of cell passages. Human embryonic lung fibroblasts (MRC-5) at passage (P) 27 were seeded twice a week to P 32, P 40, and P 48, when just at confluence and then infected with herpes simplex virus type 1 (HSV-1). The structure of the non-virus infected (MOCK) and HSV-1 infected cells, the amount of cellular infectious virus particles and the capability to express HSV-1 glycoproteins C (gC-1) and D (gD-1) were investigated by phase-contrast and immunofluorescence light microscopy, immunogold cryosection EM, plaque assays, immunoblots, and total protein assays. Modified cell structure including fragmentation of tubulin fibers were visible in MOCK from P 38 onwards. The quantity of vimentin remained unchanged while actin accumulated and beta-tubulin decreased in HSV-1 infected late P cells compared to early P cultures. Cells of high P counts contained significantly fewer infectious virus particles, very likely of lower virulence, and their expression of gC-1 and gD-1 were concordantly reduced. These observations indicate that the number of cell P must be considered in order to reproduce results of cell biology and viral morphogenesis. The MRC-5 cells ought not to be passaged more than ten times beyond P 27 in the laboratory. PMID- 10680964 TI - Use of the confocal microscope to determine polyomavirus recombinant capsid-like particle entry into mouse 3T6 cells. AB - The structural protein genes of polyomavirus were expressed in the baculovirus system, and the proteins were found to assemble into capsid-like particles capable of packaging insect cell DNA. Recombinant capsid-like particles could be produced that were composed of the various structural proteins (VP1, VP1/2, VP1/3 and VP1/2 + VP3). Laser scanning confocal microscopy was used to determine if the various capsid-like particles could infect (enter) mouse 3T6 cells. Each of the various capsid-like particles was equally capable of cell entry as determined by indirect immunofluorescence confocal microscopy. PMID- 10680965 TI - Detection of sheep poxvirus in skin biopsy samples by a multiplex polymerase chain reaction. AB - The development of a multiplex polymerase chain reaction (PCR) method with amplification of capripoxvirus in a single-step procedure from skin biopsies using three primer pairs, two specific for capripoxvirus and one specific for alpha-tubulin is described. A sensitive multiplex PCR was achieved by optimization of parameters such as the primer concentrations, magnesium and dNTPs concentrations. False negative results that sometimes arise due to inhibitors of DNA amplification may be avoided by the inclusion in the assay of alpha-tubulin primers. The results reported on 42 skin biopsies from sheep suspected to have poxvirus infection, indicated that the assay could monitor simultaneously DNA extraction from skin biopsy samples and allow improved detection of capripoxvirus within 24 h of specimen receipt in the laboratory. PMID- 10680966 TI - Serological diagnosis of varicella-zoster virus in sera with antibody-capture enzyme-linked immunosorbent assay of IgM. AB - Evaluation was made of three enzyme-linked immunosorbent assay (ELISA) formats; varicella-zoster virus (VZV) indirect ELISA; VZV IgM capture using biotin and VZV IgM capture using peroxidase, for the detection of VZV-specific IgM antibodies in human sera. It was observed that there was no significant difference in sensitivity of detection using the three formats but there were important practical differences in the number of steps and hence time for assay completion between the three assay formats. All assays showed some cross-reactivity with sera containing anti-HSV1 antibodies. PMID- 10680967 TI - Dot-blot nitrocellulose enzyme immunoassays for the detection of white-spot virus and yellow-head virus of penaeid shrimp. AB - Dot-blot nitrocellulose enzyme immunoassays (DB-NC-EIA) were developed for the detection of white-spot virus (WSV) and yellow-head virus (YHV) in infected shrimp. The assays utilized HRP-conjugated virus-specific antibodies to detect virus antigen present in gill homogenates of infected shrimp spotted onto nitrocellulose membrane. The assays are by far the simplest and most rapid detection methods available for WSV and YHV. PMID- 10680968 TI - Evaluation of newly developed microparticle enzyme immunoassays for the detection of HCV antibodies. AB - The newly developed anti-HCV assays AxSYM HCV version 3.0 and IMx HCV version 3.0 were evaluated with regard to their precision, sensitivity and specificity in comparison to the HCV EIA 3.0 (Abbott GmbH, Wiesbaden, Germany). Precision testing was undertaken using five positive controls with different anti-HCV levels for each assay. Specificity was estimated by testing 4383 blood donor specimens. The supplemental assay Matrix HCV 2.0 (Abbott GmbH, Wiesbaden, Germany) was used to confirm repeatedly reactive results. Samples which had been found to be positive or indeterminate by Matrix HCV 2.0 were tested by qualitative polymerase chain reaction after reverse transcription (RT-PCR, Amplicor HCV test, Roche Diagnostic Systems, Basel, Switzerland). To determine sensitivity, 20 commercially available seroconversion panels were tested. Based on supplemental testing, the apparent specificities of AxSYM HCV version 3.0, IMx HCV version 3.0 and HCV EIA 3.0 were estimated to be 99.84, 99.98 and 99.80%, respectively. In seroconversion panel testing, AxSYM and IMx HCV version 3.0 detected seroconversion in up to 12/20 panels earlier and in up to 1/20 cases later than the comparison EIA. The highest sensitivity was shown in AxSYM HCV version 3.0, followed by IMx HCV version 3.0 and HCV EIA 3.0. Based on the improved seroconversion sensitivity and specificity, the AxSYM and IMx HCV version 3.0 assays appear to be suitable for detecting HCV antibodies in blood donor testing and other routine laboratory assessments. PMID- 10680969 TI - Evaluation of different measles IgG assays based on recombinant proteins using a panel of low-titre sera. AB - During the WHO campaign to eradicate measles, accurate discrimination between immune and non-immune individuals will become increasingly important. Due to waning immunity in vaccinated populations, the performance of a measles IgG assay depends mainly on its ability to detect reliably seronegative individuals among many vaccinees with low antibody levels. New serological tests based on recombinant proteins detect only a fraction of the total measles virus (MV) specific antibodies. Therefore, several assays based on recombinant MV haemagglutinin (ELISA and flow cytometry) or MV-fusion protein (flow cytometry) as well as neutralisatlon and haemagglutination test have been evaluated using a large panel of low-titre and negative sera. Since such an evaluation is highly dependent on threshold values for positivity, the receiver operating characteristic curve analysis was applied. The H-FACS and the H-ELISA showed the best performing characteristics (specificity: 97.4 and 96.1%, respectively; sensitivity: 88.1 and 89.6%, respectively) and may be an alternative to the neutralisation assay. The number of undefined/grey zone sera was significantly lower compared to a commercial whole virus-based ELISA and therefore fewer individuals would be vaccinated unnecessarily. PMID- 10680970 TI - Detection and diagnosis of parapoxvirus by the polymerase chain reaction. AB - The genus Parapoxvirus includes four members, bovine papular stomatitis virus (BPSV), pseudocowpox virus (PCPV), orf virus (ORFV) and parapoxvirus of red deer in New Zealand (PVNZ). A set of primers for polymerase chain reaction (PCR) was designed to detect viral DNA from cells infected with each of the four parapoxviruses. The set of primers resulted in the amplification of appropriately sized products from cells infected with BPSV, PCPV, ORFV and PVNZ, respectively. The PCR method was applied for the detection of seven field isolates of parapoxvirus from cattle, sheep and free-ranging wild Japanese serows. The expected size of DNA was amplified from cells infected with each of the seven isolates. No specific PCR products were detected from vaccinia virus-, fowlpox virus- and mock-infected cells. Moreover, by a semi-nested PCR with an inner primer and Southern blot analysis, viral DNA was detected from lesions of clinically affected cattle, sheep and Japanese serows. These results suggested that the PCR method used in this study was specific for the detection of parapoxviruses and thus useful for diagnosis of parapoxvirus infections, especially in discrimination from diseases with similar clinical symptoms. PMID- 10680971 TI - cDNA probes for detection of specific dsRNAs from the fungal pathogen, Monosporascus cannonballus. AB - Monosporascus cannonballus is an ascomycete fungus that is the causative agent of Monosporascus root rot/vine decline, a serious disease of muskmelon and watermelon. Double-stranded RNA (dsRNA) was identified in approximately 60% of M. cannonballus isolates recovered from infected muskmelon plants in 1993. After repeated laboratory transfer on culture media, the majority of the isolates harboring dsRNAs developed degenerate culture phenotypes and showed reduced virulence (hypovirulence) to muskmelon. Initially, dsRNA purification and cDNA synthesis were attempted in three M. cannonballus isolates harboring dsRNAs. However, numerous difficulties were encountered due to the stable, double stranded nature of the dsRNAs and contamination of the preparations by fungal rRNA. Several purification and cDNA protocols were evaluated and eventually modified into methods that were ultimately highly effective for cloning dsRNAs from M. cannonballus. The cDNAs derived from purified dsRNA preparations were cloned into a pUC119 plasmid vector and amplified in Escherichia coli. Nine cDNA clones were identified that are specific for medium-sized (ca. 3 kbp) dsRNAs associated with M. cannonballus isolate Ca91-17(96+). The methods used to make the cDNA clones of the dsRNAs in M. cannonballus may be useful for those working on fungal dsRNAs. In addition, these cDNAs may be useful for identifying dsRNAs associated with the hypovirulence phenotype. PMID- 10680972 TI - Folate-mediated drug delivery: effect of alternative conjugation chemistry. AB - When utilized as a macromolecular drug targeting ligand, folic acid (Pte-Glu) has traditionally been coupled to peptides, proteins and lipids via one of its two carboxylate groups fortuitously located within a distal glutamyl moiety. It has been assumed in the literature that the gamma-glutamyl carboxylate of Pte-Glu is the preferred conjugation site for macromolecules enduring endocytosis via the folate-binding protein receptor. However, it is also possible that the steric placement of the attached macromolecule around the vitamin's pteridine moiety may be the more influential parameter controlling this delivery mechanism. Using solid-phase chemistries, we have synthesized dipeptide derivatives of pteroic acid for the purpose of identifying the preferred site onto which a macromolecule can be chemically attached without compromising its endocytosis potential. Thus, using fluorescent and radiolabeled conjugates, we have determined that macromolecules attached to Pte-Glu by either an alpha- or gamma-glutamyl linkage could associate with receptor-bearing cells at virtually identical levels. We further discovered that removal of the remaining un-conjugated glutamyl carboxylate had no inhibitory effect on cell uptake; and, the cytotoxicity of related momordin toxin conjugates were comparable among the various pteroate derivatives tested. From these observations we suggest that the preparation of endocytosis-competent pteroate-macromolecule conjugates is strongly influenced by the steric environment around the ligand's para-aminobenzoic acid moiety, and that no selective isomeric (i.e. alphaGlu versus gammaGlu) conjugation requirement necessarily exists. PMID- 10680973 TI - Selective delivery of adriamycin to a solid tumor using a polymeric micelle carrier system. AB - The anticancer drug, adriamycin (ADR), was incorporated by physical entrapment into polymeric micelles for selective delivery to a murine solid tumor colon adenocarcinoma 26 (C 26). In vivo antitumor activity of ADR was greatly enhanced by this incorporation into polymeric micelles. Using one polymeric micelle delivery system, the tumor completely disappeared at two doses, while free ADR exhibited a fair inhibition effect on tumor growth only at the maximum tolerated dose. Biodistribution analysis revealed that the physically entrapped micellar ADR accumulated at tumor sites in a highly selective manner. These results indicate that these polymeric micelles are a promising system for delivering hydrophobic anticancer drugs selectively to solid tumor sites using a passive targeting mechanism. PMID- 10680974 TI - Lipid microsphere preparation of a lipophilic ceramide derivative suppresses colony formation in a murine experimental pulmonary metastasis model. AB - Ceramide is a well-known regulator of apoptosis and cell growth. In this study, we synthesized lipophilic ceramide derivatives to incorporate into lipid microspheres (LM) and their activity was evaluated in vivo. Cera 03, a lipophilic ceramide derivative synthesized from membrane-permeable C2-ceramide, caused potent growth inhibition and DNA fragmentation of Meth A-T tumor cells in vitro. Its potency was similar to that of C2-ceramide. Both compounds increased the proportion of apoptotic cells. Cera 02, the diacetylated form of natural ceramide (Cer), also suppressed in vitro cell growth with a similar or higher potency to that of Cer, but both were far less potent than C2-ceramide and Cera 03. LM containing Cera 03 (Lipo-Cera 03) could not totally prevent metastatic incidence of Meth A-T cells, but reduced pulmonary metastatic nodules in number. Intravenous injection of Lipo-Cera 03 (1 mg/kg of Cera 03) produced about 35% inhibition, while Lipo-Cera 02 had no significant effect. In conclusion, Lipo Cera 03 may have potential as an antimetastatic drug and may also be a useful tool for researching the role of ceramides in vivo. PMID- 10680975 TI - Effects of methotrexate and cyclophosphamide on polyamine levels in various tissues of rats. AB - The effects of methotrexate (MTX) and cyclophosphamide (CYP) on body weight, organ weight, and the concentration of putrescine, spermidine, and spermine in 14 different tissues were measured in rats that had been given these compounds for 5 consecutive days. These three polyamines in both the thymus and spleen of rats treated with MTX and CYP showed a statistically significant decrease. Further, putrescine in the seminal vesicles, kidney, liver, and small intestine of MTX treated rats, and in the prostate, seminal vesicles, kidney, heart, liver, small intestine, and lung of CYP-treated rats, spermidine in the prostate, seminal vesicles, testis, thymus, spleen, kidney, heart, small intestine, and skeletal muscle of CYP-treated rats, and spermine in the prostate, seminal vesicles, kidney, heart, small intestine, and stomach of CYP-treated rats showed statistically significant decreases. Recognition of the significance of polyamine levels and attention to their response in anti-cancer drug therapy may have clinical implications. PMID- 10680976 TI - Pre-exposure of cells to cationic lipids enhances transgene delivery and expression in a tissue culture cell line. AB - Several factors influence non-viral transfection in tissue culture models including nature of the cationic lipid, plasmid construction, and DNA lipid complex, among others. The cell line itself is another confounding variable. Each subcellular population may respond independently to the transgene or specific delivery vector with regards to toxicity or transgene expression. In this study, the SKnSH (human neuroblastoma) and COS-1 (African green kidney) cells were exposed to three different treatments A, B, and C. Treatment A refers to cells obtained from American Type Culture Collection (ATCC) and cultivated as recommended, treatment B to cells that were grown in presence of cationic lipids for two weeks, and treatment C to cells that were grown in presence of cationic lipids for two weeks followed by normal media for two weeks to determine if lipid mediated effects were reversible. Treatment B resulted in a three-fold increase in transgene expression of a reporter gene as compared to the other treatments. This increase in transgene expression appeared not to be related to alterations in toxicity. Interestingly, the fluid phase endocytic uptake of fluorescently labeled oligonucleotides was increased in treatment B. However, there was no significant difference in the cellular-associated signal when fluorescently labeled plasmid-DNA was evaluated. In COS-1 cells, no difference in transfection was observed with treatment B illustrating that cell lines respond independently. In conclusion, pre-exposure of SKnSH cells to cationic liposomes (treatment B) resulted in higher transgene production. PMID- 10680977 TI - Uptake by hepatocytes and biliary excretion of intravenously administered polystyrene microspheres in rats. AB - The in vivo uptake by hepatocytes and biliary excretion of fluorescein isothiocyanate-labeled polystyrene microsphere with a particle size of 50 nm (MS 50) after intravenous administration was studied in rats. It was confirmed by using confocal laser scanning microscopy that MS-50 was partially phagocytosed by the hepatocytes and that MS-50 taken up by the hepatocytes existed exclusively inside the cells 1 h after intravenous administration. Studies on the mechanism of the uptake of MS-50 by the hepatocytes using the liver perfusion technique revealed that a process mediated by apo-E was involved. After intravenous administration of MS-50, about 4% of dose was excreted into bile in 24 h. Pharmacokinetic evaluation of the excretion rate of MS-50 into bile showed that the process followed first-order kinetics. Qualitative evaluation of the fluorescence detected in the bile after intravenous administration of MS-50 revealed that the particles were certainly excreted into bile in an intact form. From these results, it was suggested that intravenously administered MS-50 would be partially phagocytosed by hepatocytes through a process mediated by apo-E and that MS-50 ingested by hepatocytes would be partially excreted into the bile. PMID- 10680978 TI - Characterization of norfloxacine release from tablet coated with a new pH sensitive polymer, P-4135F. AB - A new pH-sensitive polymer, P-4135F, was evaluated as a colon delivery device for norfloxacine (NFLX) which is used for the therapy of patients with Vero toxin producing Escherichia coli gastroenteritis. P-4135F has a dissolution threshold pH of 7.2 which is higher than the conventional pH-sensitive polymers, Eudragit S100 and L100. To compare the dissolution site of P-4135F coated tablets with other enteric polymer coatings, mini-tablets containing sodium fluorescein (FL) as a model drug were prepared by coating them with the three polymers. After oral administration of FL mini-tablets to rats, the first-appearance time, Ti, of FL into the systemic circulation was measured. The Tis were 0.7+/-0.2 h for Eudragit L100, 1.8+/-0.4 h for S100 and 2.0+/-0.3 h for P-4135F. Direct inspection of the dissolution process of the FL mini-tablets after oral administration to rats was performed by abdominal incision studies. All of the coated FL mini-tablets started to dissolve in the rat ileum. The dissolution sites were identified to be proximal to the ileocecal junction for P-4135F, at the middle part of the ileum for Eudragit S100 and at the proximal part of the ileum for Eudragit L100. NFLX tablets with different membrane thicknesses of P-4135F were prepared and were orally administered to beagle dogs. The colon delivery efficiency was evaluated by measuring the Ti of NFLX into the systemic circulation. The mean Tis were 1.33+/-0.33 h for 56.8+/-0.5 microm membranes, 3.75+/-0.25 h for 64.6+/-0.7 microm membranes, 4.00+/-1.00 h for 70.5+/-0.5 microm membranes and 3.00+/-1.00 h for 74.9+/-0.4 microm membranes. By comparing the Ti, 4.33+/-0.33 h, obtained after oral administration of NFLX in a pressure-controlled colon delivery capsule, and the colon arrival time, 3.5+/-0.3 h, determined by a sulfasalazine test in beagle dogs. P-4135F coated NFLX tablets appeared to dissolve and disintegrate before reaching the colon. Studies using rats and beagle dogs have suggested that P-4135F dissolves in the lower part of the small intestine, i.e., the ileum. These studies also suggest that this new polymer will be useful for the delivery of NFLX to the lower part of the small intestine. PMID- 10680979 TI - Transnasal delivery of 5-fluorouracil to the brain in the rat. AB - The purpose of this research is to clarify the feasibility and to determine the extent of transnasal drug delivery to the brain through the cerebrospinal fluid (CSF) in the rat, using 3H-5-fluorouracil (5FU) as a model drug. It was confirmed first that the concentration of 5FU in the CSF was significantly higher following nasal administration compared with intravenous injection, indicating direct transport of 5FU from the nasal cavity to the CSF. Concentration-time profiles of 5FU in the plasma and in the cerebral cortex were determined following intravenous infusion, nasal instillation and nasal perfusion. In order to evaluate the extent of drug transport from the nasal cavity to the cerebral cortex by way of the CSF, the apparent brain uptake clearances were calculated. The uptake clearance following nasal perfusion (8.65 microl/min/g tissue) was significantly large (p < 0.001) in comparison with that following intravenous infusion (6.20 microl/min/g tissue), while that following nasal instillation (6.94 microl/min/g tissue) was not. Consequently, significant amount of 5FU is transported from the nasal cavity to the brain through the CSF and thus, the delivery of the hydrophilic drug to the brain is augmented by nasal drug application. PMID- 10680980 TI - Pathogenic mechanism of acute post-streptococcal glomerulonephritis. AB - Considerable knowledge has been accumulated regarding the characteristics of acute post-streptococcal glomerulonephritis (APSGN), and many attempts have been made to identify a streptococcal factor or factors responsible for triggering this disease. However, the pathogenic mechanism behind APSGN remains largely unknown. As glomerular deposition of C3 is generally demonstrated before that of IgG in the disease process, it is likely that the inflammatory response is initiated by renal deposition of a streptococcal product, rather than by deposition of antibodies or pre-formed immune complexes. During recent years, a number of streptococcal products have been suggested to be involved in the pathogenic process. In this review, possible roles of these factors are discussed in the context of the clinical and renal findings most often demonstrated in patients with APSGN. Streptokinase was observed to be required in order to induce signs of APSGN in mice, and a number of findings suggest that the initiation of the disease may occur as a result of renal binding by certain nephritis associated variants of this protein. However, additional factors may be required for the development of the disease. PMID- 10680981 TI - Outbreak of echovirus 30 meningitis in Switzerland. AB - This study includes 80 patients (38 children and 42 adults) who contracted aseptic meningitis in the summer of 1996 in Fribourg, Switzerland. Virological studies revealed an enteroviral infection in 65 out of 70 (93%) investigated patients. In 47 out of the 53 cases (89%) where a precise virus could be identified, the causative agent was an Echovirus 30. More than 50 patients lived in an area within a 5-km radius. The patients presented with the classic symptoms and signs of aseptic meningitis. In contrast, polymorphonuclear leukocytes predominated in the cerebrospinal fluid in the first 24 h and 32% of the cases had a left shift in their peripheral blood smear. The patients' age did not influence white blood cell count, the proportion of polymorphonuclear leukocytes or protein concentration in the cerebrospinal fluid. Thirty-three patients (41%) received antibiotic treatment, and 38 patients (48%) left the hospital within 24 h. Only 2 neuroradiological procedures and 1 electroencephalographic recording were performed. The outcome was favourable in all patients. PMID- 10680982 TI - Acyclovir for treatment of infectious mononucleosis: a meta-analysis. AB - A meta-analysis of 5 randomized controlled trials (RCT), involving 339 patients with acute infectious mononucleosis (IM) treated with acyclovir (ACV) was performed. ACV was given intravenously in 2 RCTs, which included patients with more severe disease, and orally in the remaining 3 RCTs, which included patients with mild to moderate IM. Both clinical and virological endpoint data available from RCT were evaluated in this study. There was a trend towards clinical effectiveness of ACV treatment, but no statistically significant results were achieved. In contrast, a significant reduction in the rate of oropharyngeal EBV shedding was observed at the end of the therapy (overall OR: 6.62; 95% CI: 3.56 12.29; p < 0.00001). However, no difference in EBV shedding was observed 3 weeks later. There was no significant difference on adverse events in the groups of patients treated with ACV or placebo. In conclusion, clinical data do not support use of ACV for the treatment of acute IM, despite good virological activity of this drug. There is a need for more effective treatment of EBV infection. PMID- 10680983 TI - Cytomegalovirus UL97 and glycoprotein B (gB) sequences in tissues from immunocompromised patients with ganciclovir-resistant virus infection. AB - The pathogenesis of ganciclovir-resistant cytomegalovirus (CMV) was investigated by analysing UL97 and gB sequences in tissues obtained from 4 immunocompromised patients with infections caused by ganalciclovir-resistant virus. UL97 and gB sequences were obtained by automated sequencing of CMV DNA amplified from lysates prepared from deparaffinized tissue sections. Patient 1 contained wild-type UL97 and gB3 sequences. Patient 2 harboured genetically distinct viruses in his lung: one with a ganciclovir-resistance UL97 mutation and a gB3 genotype, and another without UL97 mutations and a gB1 genotype. In patient 3, a ganciclovir-resistant UL97 mutant virus with a gB1 genotype was cultured from the lung, whereas the CMV in the brain did not contain mutations and its genotype was gB2. In patient 4, ganciclovir-resistance UL97 sequences were found in oesophageal tissue prior to the isolation of a ganciclovir-resistant CMV from the blood. All viruses in this patient had a gB3 genotype. CMV containing ganciclovir-resistance UL97 mutations may cause end-organ disease in immunocompromised individuals. In these subjects, CMV circulating in the blood may have similar or different UL97 and gB genotypes than the virus causing end-organ disease. PMID- 10680984 TI - Inter- and intra-species karyotype variations among microsporidia of the genus Encephalitozoon as determined by pulsed-field gel electrophoresis. AB - Disseminated infections due to microsporidia of the genus Encephalitozoon are detected increasingly, especially in patients with AIDS. Identification of microsporidia can be achieved by a variety of immunological and molecular methods. This study evaluates the feasibility of pulsed-field gel electrophoresis (PFGE) for the analysis of karyotypes of the 3 known species of this genus (Encephalitozoon cuniculi, Encephalitozoon hellem and Encephalitozoon intestinalis) and of 2 of the 3 known E. cuniculi strains (strains I and III). Eleven chromosomal DNA bands were resolved for E. cuniculi and 10 chromosomal DNA bands for E. hellem and E. intestinalis, with molecular sizes ranging from 231 to 320 kb, from 197 to 288 kb and from 195 to 285 kb, respectively, resulting in estimated genome sizes of about 3.0 Mb, 2.5 Mb and 2.4 Mb. Different PFGE chromosomal banding patterns indicate that not only E. cuniculi, as previously described, but also E. hellem, represent a heterogeneous entity. PFGE is a valuable method of evaluating inter- and intra-species variations among Encephalitozoon species that may enable the identification of environmental sources of infection and modes of transmission. PMID- 10680985 TI - Bacterial arthritis in a Swedish health district. AB - The aim of this retrospective investigation was to study the occurrence, characteristics and outcome of culture-positive bacterial arthritis in a Swedish county hospital for the period 1994-97. Using registers of diagnosis, 15 adult patients with infectious arthritis in native joints were identified. Staphylococcus aureus was, as expected, the most common microbial agent. Six cases were caused by direct inoculation. The risk for bacterial arthritis caused by intra-articular steroid injection was estimated at 1/12,000. Joint damage was seen in 6 cases. Two patients died during the infection. White blood cell count in synovial fluid was performed in 5 cases, with median value 1.9 x 10(9)/l and, since it was not a good predictor of infection in this study, the value of this method for this indication is questioned. PMID- 10680986 TI - Tuberculosis diagnosed in a surgical department during 1974-93: a report of 32 cases. AB - During the period 1974-93, 32 patients (18 males, 14 females) with previously undiagnosed tuberculosis were admitted to the department of surgery and the surgical outpatient department of Buskerud Central Hospital. Of these, 26 were native Norwegians and 6 immigrants of Asian origin with an average age of 65 and 26 years, respectively. Of the native Norwegians, 14 patients had a past history of tuberculosis and 7 cases were associated with concomitant cancer. With 1 exception, all immigrants had extrapulmonary tuberculosis. Cultural identification of Mycobacterium tuberculosis occurred in 28 cases and Mycobacterium bovis in 1 (immigrant). A total of 30 patients were cured, whereas 2 of 3 patients with miliary dissemination died. PMID- 10680987 TI - Trovafloxacin susceptibility of aerobic clinical bacterial isolates from Sweden. AB - Trovafloxacin susceptibility was studied in aerobic clinical isolates of bacterial pathogens from 5 microbiology laboratories in Sweden. Trovafloxacin and ciprofloxacin minimum inhibitory concentration (MIC) determinations were performed on 474 clinical isolates. Disk diffusion tests using trovafloxacin and ciprofloxacin 10 microg disks were performed on a total of 7142 clinical isolates (trovafloxacin). Susceptibility interpretations for trovafloxacin and ciprofloxacin were determined from MIC values and disk diffusion tests using species-related MIC-limits and zone diameter breakpoints. Eight of 12 gram positive species groups were fully susceptible to trovafloxacin as judged by MIC tests. Trovafloxacin gave MIC50 values of 0.032 mg/l for S. aureus, 1.0 mg/l for MRSA, 0.064 mg/l for coagulase negative staphylococci, 1.0 mg/l for MRSE, 0.064 mg/l for S. saprophyticus, 0.125 mg/l for group A and group B streptococci, 0.064 mg/l for group C and G streptococci and S. pneumoniae, 0.25 mg/l for E. faecalis, and 16.0 mg/l for E. faecium. These MIC values were 4-16-fold lower than those of ciprofloxacin. Both MIC and disk tests showed similar levels of susceptibility among gram-negative isolates for trovafloxacin and ciprofloxacin. For most gram negative species the trovafloxacin MIC50 values were similar to or slightly higher than those for ciprofloxacin. Trovafloxacin MIC values were much lower for Acinetobacter strains, but higher for P. mirabilis compared with ciprofloxacin. The favourable susceptibility levels of Swedish aerobic pathogens to trovafloxacin emphasize the potential of this drug for the treatment of serious infections. PMID- 10680988 TI - Calibration of disk diffusion antibiotic susceptibility testing: species-related trovafloxacin interpretive zone breakpoints and selection of disk potency. AB - International comparisons of antibiotic susceptibility require the use of common minimum inhibitory concentration (MIC) limits. Disk diffusion test results are not directly suitable for such comparisons, since different standards are often used and zone breakpoints issued might reflect different MIC limits. We have used single strain regression analysis (SRA) for the calibration of the disk test, both according to species and individual laboratory, and for quality control of trovafloxacin disk diffusion tests in 5 laboratories in Sweden. Preliminary controls using histogram analysis including subtraction histograms of reference strains revealed marked differences between different laboratories. SRA was performed on 4 reference strains, S. aureus, E. faecalis, E. coli and P. aeruginosa, using disks containing 1, 3, 10, 30 and 100 microg trovafloxacin. The results using SRA showed a difference between laboratories using Biodisk PDM medium, which produced smaller zones, and those using Oxoid IsoSensitest. Species related regression lines for laboratories using either medium were calculated and corresponding interpretive zone breakpoints determined for MIC limits. Rational criteria for the selection of a suitable disk content of an antibiotic were also defined and applied to trovafloxacin. The 10 microg disk selected by NCCLS (National Committee for Clinical Laboratory Standards) proved optimal. PMID- 10680989 TI - Immunogenicity and safety of a tetravalent diphtheria-tetanus-acellular pertussis inactivated poliovirus vaccine. AB - The objective of this study was to investigate whether a tetravalent vaccine containing diphtheria, tetanus, monocomponent acellular pertussis and inactivated poliovirus (DTaP-IPV) was immunogenic and safe compared with the vaccination regime used in Denmark at the time of the study. The study was performed as an open controlled study in which 270 Danish children were enrolled at their 5 weeks' routine examination. The children were allocated to receive either (i) DTaP-IPV (12.5 Lf, 7 Lf, 40 microg, 40, 8, 32 DU) at 3, 5 and 12 months of age (n = 186) or (ii) DT-IPV (50 Lf, 12.5 Lf, 40, 8, 32 DU) at 5, 6 and 15 months of age plus whole-cell pertussis vaccine (> or = 4 IU) at 5 and 9 weeks and at 10 months of age (n = 84). No hypotonic hyporesponsive episodes or other vaccine-related serious adverse events were seen. Local reactions, febrile and crying episodes with the investigational vaccine (DTaP-IPV) were similar to the reactions seen with the existing DT-IPV vaccine. One month after completing the vaccination schedule, all children had antibodies above the defined protective antibody concentrations to polio, tetanus and diphtheria. For pertussis toxin, there was a significantly better response in the investigational vaccine group. We therefore conclude that, when used according to the schedule tested, the tetravalent DTaP IPV vaccine is safe and immunogenic. In addition, the number of visits and the number of injections necessary are reduced with this vaccine and vaccination schedule. PMID- 10680990 TI - Safety and immunogenicity of combined Dtpa-IPV vaccine for primary and booster vaccination. AB - This study evaluated the safety and immunogenicity of combined diphtheria, tetanus, acellular pertussis, inactivated poliomyelitis vaccine (DTPa-IPV) given as primary immunization at ages 3, 4.5 and 6 months and a booster dose between the ages of 18 and 27 months to healthy children. The acellular pertussis tricomponent vaccine contains pertussis toxoid (PT), filamentous haemaglutinin (FHA) and 69 kDa outer membrane protein (PRN). Serum immune responses to the administered antigens were measured before and after the primary and the booster vaccination series. The safety of the vaccine was evaluated based on diary cards completed by parents within 4 d following each vaccination. A total of 237 and 150 children completed the primary and booster vaccination series, respectively. A total of 483 (66.5%) and 111 (74%) local and 317 (43.7%) and 98 (65.3%) general adverse events were reported after 726 doses of the primary series and 150 of the booster doses, respectively. Compared with primary vaccination, the incidence of all adverse symptoms was greater after the booster dose and a previous severe reaction was a risk factor for a severe reaction after the booster dose (OR = 5.11). All but 1 child, who failed to have antibodies to diphtheria toxoid after the booster dose, responded to all administered antigens with antibody titres greater than the assay cut-off points. The combined DTPa-IPV used for primary and booster immunization induced good immunity, but was associated with large local reactions in 21.3% of children after the booster dose. PMID- 10680991 TI - Subacute primary Candida lung abscess. AB - A case of primary subacute Candida lung abscess is described. The most reliable way to diagnose a rare pulmonary disease is to perform an open lung biopsy. A review of the literature suggests that the diagnosis of a primary subacute abscess due to Candida albicans in vivo is unique. PMID- 10680992 TI - Community-acquired Staphylococcus epidermidis endocarditis complicated by splenic disease 9 years after aortic valve replacement. AB - We describe here a case of Staphylococcus epidermidis prosthetic aortic valve endocarditis 9 y after valve replacement. Surprisingly, the microorganism was community-acquired, highly virulent and associated with splenic disease. The patient recovered following emergency valve replacement and prolonged antibiotic therapy. Splenectomy was not required. PMID- 10680993 TI - Endocarditis caused by drug-resistant Streptococcus pneumoniae in a child. AB - We report a case of infective endocarditis caused by drug-resistant Streptococcus pneumoniae. Cefazolin or cefotaxime therapy induced a partial response. Treatment with vancomycin was successful. This microorganism may be more significant in endocarditis in areas with a high prevalence of drug-resistant Streptococcus pneumoniae. PMID- 10680994 TI - Streptococcus equinus endocarditis in a patient with pulmonary histiocytosis X. AB - Although Streptococcus equinus has been isolated from the human bowel in an appreciable percentage of the adult general population, it has only rarely been described as a human pathogen. Our report describes the occurrence of S. equinus endocarditis in a patient who had no history of pre-existing heart disease, but who showed evidence of a late-stage pulmonary histiocytosis X. Endocarditis resolved promptly after antibiotic treatment, but required aortic valve substitution. Abnormalities of the immune system that have been demonstrated in patients with histiocytosis X could explain the occurrence of endocarditis in this patient. PMID- 10680995 TI - Leukocyte adhesion deficiency in a Norwegian boy. AB - A Norwegian boy suffering from recurrent urinary and respiratory tract infections was examined. It was found that granulocyte random migration, respiratory burst activity and the proportions of CD11/CD18 receptor positive leukocytes were reduced, consistent with a diagnosis of leukocyte adhesion deficiency. An increased proportion of Fc gammaI-receptor-bearing granulocytes did not compensate for the CR3 deficiency. PMID- 10680996 TI - Praziquantel in niclosamide-resistant Taenia saginata infection. PMID- 10680997 TI - Loss of livestock breeding efficiency due to uncompensable sperm nuclear defects. AB - An important goal of modern analyses of semen is to elucidate the molecular traits of mammalian sperm chromatin structural abnormalities, defined here as 'uncompensable', that lead to abnormalities in fertility, pronuclear formation, early embryo quality and pregnancy outcome. Sperm with uncompensable nuclear abnormalities are able to fertilize oocytes both in vivo and in vitro; however, due to the uncompensable trait(s), the embryo development may be abnormal. Uncompensable nuclear traits can be experimentally induced in bull sperm by a mild thermal insult to the testis. Sperm nuclear morphology abnormalities seen in ejaculates 11-days post stress are likely related to molecular changes in chromatin observed 3-days post stress by the flow cytometric sperm chromatin structure assay (SCSA). The SCSA measures the susceptibility of sperm nuclear DNA to denaturation in situ. This susceptibility has been correlated with the presence of DNA strand breaks that may be derived in part by oxidative stress and possibly by a unique, abortive apoptotic mechanism. The extent of DNA denaturation is not significantly related to the level of disulfide bonding between the chromatin protamines. The use of human sperm with uncompensable nuclear traits for artificial reproductive techniques is also discussed. The goal of this research is to remove from semen doses those sperm with uncompensable nuclear traits and thereby increase male fertility potential. PMID- 10680998 TI - Molecular cloning of translocation breakpoints in a case of constitutional translocation t(11;22)(q23;q11) and preparation of probes for preimplantation genetic diagnosis. AB - In vitro fertilization (IVF) centres with preimplantation genetic diagnosis (PGD) programmes are often confronted with the problem of identifying chromosomal abnormalities in interphase cells biopsied from preimplantation embryos of carriers of a reciprocal translocation. The present authors have developed a DNA testing based approach to analyse embryos from translocation carriers, and this report describes breakpoint-spanning probes to detect abnormalities in cases of the most common human translocation (i.e. the t(11;22)(q23;q11)). Screening a yeast artificial chromosome (YAC) library for probes covering the respective breakpoint regions in the patient lead to probes for the breakpoint on chromosome 11q23. The physically mapped YAC and bacterial artificial chromosome (BAC) clones from chromosome 22 were then integrated with the cytogenetic map, which allowed localization of the breakpoint on chromosome 22q11 to an interval of less than 84 kb between markers D22S184 and KI457 and to prepare probes suitable for interphase cell analysis. In summary, breakpoint localization could be accomplished in about 4 weeks with additional time needed to optimize probes for use in PGD. PMID- 10680999 TI - Kinematic definition of ram sperm hyperactivation. AB - Although it is known that ram spermatozoa exhibit hyperactivated motility under capacitating conditions, quantitative analyses of the head and flagellar movement of washed ram spermatozoa have not been published. Motile spermatozoa were recovered from semen by swim-up into HSOF medium, and their movement in 30-microm deep chambers was videorecorded. Spermatozoa of interest were identified during tape playback (hyperactivated spermatozoa were identified by visual assessment of flagellar movement) and sequential head and tail images were traced onto overhead projector film attached to the video monitor. The flagellar movement characteristics beat angle (FBA), beat envelope (FBE) and curvature ratio (FCR) were determined by first principles, and head centroid kinematics were determined using Cartesian methods. Hyperactivated spermatozoa had significantly higher FBA and FBE and significantly lower FCR values than non-hyperactivated spermatozoa (all P<0.0001). The centroid kinematic values were also found to be significantly different, and kinematic criteria for ram sperm hyperactivation were developed. These criteria were refined by consideration of 60-Hz CASA-derived trajectories, and ram sperm hyperactivation was defined by: VCL > 250.0 microm s(-1) and VSL < or = 100.0 microm s(-1) and LTN < or = 30% and ALHmax > or = 9.0 microm. PMID- 10681000 TI - Expression of polo-like kinase (PLK) in the mouse placenta and ovary. AB - The polo-like kinase (PLK) is a mammalian serine/threonine kinase involved in cell cycle regulation. Much evidence for the role of PLK in the cell cycle has come from studies of cultured cells; however, little is known about its function or even expression in vivo. The present study examined the features of PLK expression in the mouse placenta and ovary. Immunohistochemical studies showed that PLK is highly expressed in the basement membrane of the endometrial gland, in some endothelial cells, in endometrium after embryo implantation, in trophoblastic tissue invading the decidua, in the ovarian stroma and in some lutein bodies. In contrast, PLK was not detectable by immunohistochemistry in endometrial stroma before decidualization, in decidua, in trophoblastic tissue not invading the decidua or in ovarian follicles. PLK expression seemed to be correlated with the expression of proliferation cellular nuclear antigen (PCNA) in many placental and ovarian cells, reflecting a role in cellular proliferation. Nevertheless, in ovarian stroma and lutein bodies where PCNA was not expressed, PLK was strongly expressed. This finding indicates that PLK may have some post mitotic functions in certain specialized cell types. PMID- 10681001 TI - Development of the membrana granulosa of bovine antral follicles: structure, location of mitosis and pyknosis, and immunolocalization of involucrin and vimentin. AB - The membrana granulosa of the ovarian follicle is termed the 'follicular epithelium', yet there have been no studies considering its epithelial nature and how it changes during follicular development. Therefore, these issues were investigated using histology (n = 45 ovaries), considering its structure and the location of proliferating and dying cells, and drawing analogies with other epithelia. Additionally, differences between the layers of granulosa cells were demonstrated by immunohistochemistry (n = 7 ovaries). The structure of the membrana granulosa differed between follicles. Six arbitrary classifications were designed based on these structures, 80 follicles were allocated (n = 13 ovaries) to these classes and the follicular diameters were then measured. For the first time, differences in membrana granulosa structure were shown to correspond to follicle size. Follicles in classes 1-3, where basal granulosa cells were columnar with nuclei positioned basally in the cell, were all < or = 3 mm in diameter. All follicles larger than 3 mm had either columnar basal cells with nuclei positioned centrally (class 4), or had rounded basal cells (class 5), and all follicles > 5 mm had only rounded basal cells. In all these classes, cells in the middle zone were rounded; cells aligning the antrum were often flattened. Irrespective of follicle class, cell proliferation and cell death were shown to be predominantly in the middle portions, rather than the most antral or most basal portions, of the membrana granulosa of healthy and atretic follicles. Involucrin, a marker of keratinocyte differentiation, was localized to the suprabasal region of the membrana granulosa of healthy follicles, particularly in the second and third cellular layers in from the follicular basal lamina. Conversely, the staining intensity for the intermediate filament protein vimentin was lowest in this region, and greatest in the more antral and basal regions. In atretic follicles, there was widespread staining for involucrin and vimentin throughout the membrana granulosa. In conclusion, the membrana granulosa is highly structured, and alters with follicular development. Layers in the membrana granulosa can differ in terms of cell shape, and differ in proliferation and gene expression. In the light of the current work, and an associated study, it is proposed that proliferation occurs in the middle layers, and that granulosa cells, then progress basally or antrally, the latter undergoing terminal differentiation. PMID- 10681002 TI - Sperm antibodies in rat models of male hormonal contraception and vasectomy. AB - The presence of sperm antibodies correlates with nearly every pathological condition of the male reproductive tract. In the seasonal breeder, mink, a decrease in gonadotrophin secretion and testicular regression also induces sperm antibodies. Because the Sertoli cells and the principal cells of the epididymis (i.e. the cells mainly responsible for protection of germ cells from autoimmune destruction) are dependent on androgens, and because the androgen concentration decreases in both the testis and epididymis during male hormonal contraception, the presence of IgG class sperm antibodies in serum was studied in rats during the suppression and recovery phases of testosterone contraception and after vasectomy. Five-centimetre long testosterone implants were placed under the dorsal skin of rats under pentobarbitone anaesthesia. The control rats received empty implants. All implants were left in the rats for 27 or 53 days. The total number of testicular antigens detected by sera from the vasectomized rats increased significantly until 66 days post-operation, and then decreased to the levels of intact rats. The number of testicular antigens detected by sera from rats receiving contraceptive doses of testosterone did not increase before the testosterone capsules were removed, but at 40 days post removal of the silastic capsules, the number of antigens detected by the sera was significantly higher than in intact rats and at 77 days post removal of the silastic capsules, the number of antigens detected by the sera was significantly higher than at 27 days after starting testosterone administration. No significant changes in the number of antigens detected by the sera could be observed after the implanting of empty capsules or after their removal. Vasectomy mostly induced antibodies against testicular antigens in the molecular ratio ranges of 70-82, 25-33 and 21-24.5 kD. Antibodies against antigens in these molecular ratio ranges were not significantly induced during or after treatment with contraceptive doses of testosterone. Cell nuclei with apoptotic morphology could be observed in the seminiferous tubules of the vasectomized rats, but DNA in situ 3'-end labelling of testes could not confirm any differences between the testes of vasectomized and sham-operated rats or between testosterone-treated and empty implant-treated rats. CD3+ T cells could not be observed in the testes of any of the treatment groups. These results suggest that the immunological conditions remain stable in the testes after vasectomy and during testosterone treatment, but that the animals are more prone to develop autoantibodies after vasectomy and during recovery from treatment with exogenous testosterone. PMID- 10681003 TI - Effects of bull, sperm type and sperm pretreatment on male pronuclear formation after intracytoplasmic sperm injection in cattle. AB - This study investigated the effects of the bull, sperm type (dead, immotile or motile) and sperm pretreatment (i.e. mechanical (tail-cutting or tail-scoring) or chemical (heparin, heparin + caffeine, calcium ionophore A23187 or dithiothreitol)) on male pronuclear formation after intracytoplasmic sperm injection (ICSI) in cattle. Three experiments were conducted. In Experiment 1, spermatozoa from three bulls (A, B and C) were used for both ICSI and in vitro fertilization (IVF). The results were that sperm from bull B yielded a higher penetration/male pronuclear formation rate than that of bull C when used for IVF (89.6% v 25.6%, P<0.01). However, when injected into oocytes by ICSI, sperm from bull C had a higher male pronuclear formation rate than that of bull B (34.6% v. 16.1%, P<0.05). The effects of sperm type and mechanical pretreatment were examined in Experiment 2. No significant difference was found in the male pronuclear formation rate when the three types of sperm were injected into oocytes. Tail-scored sperm achieved a higher male pronuclear rate than that of non-mechanically treated ones (38.2% v. 13.2%, P<0.005). In Experiment 3, chemical pretreatments were tested and compared. Higher male pronuclear rates, compared with the control, were obtained when sperm were pretreated with heparin + caffeine, calcium ionophore A23187 and dithiothreitol (48.2%, 62.5% and 64.5% v. 25.0%, P<0.05, 0.005 and 0.005, respectively). These results indicate that (1) there is a bull variation in male pronuclear formation with ICSI, and the male pronuclear rate by ICSI is not coincident with the results by IVF, (2) immobilization of a spermatozoon by tail-scoring before ICSI can improve the formation of the male pronucleus, and (3) an appropriate chemical pretreatment of spermatozoa is necessary to achieve a higher rate of male pronuclear formation. PMID- 10681004 TI - Effects of pre- and post-mating nutritional status on hepatic function, progesterone concentration, uterine protein secretion and embryo survival in Meishan pigs. AB - This experiment examined whether the pre- or the post-mating diet had greater impact on embryo survival in Meishan gilts. Gilts received either a maintenance (1.15 kg day(-1); n = 12) or a high (3.5 kg day(-1); n = 12) diet during the oestrous cycle preceding mating. After mating, half the animals in each group received either the maintenance or the high diet until slaughter on Day 12. Gilts fed the high pre-mating diet had more corpora lutea (22.7 v. 19.0, SED = 0.98; P<0.001), increased embryo survival (95.5% v. 74.8%, SED = 7.58; P<0.01) and heavier corpora lutea (-0.71 log g v. -0.90 log g, SED = 0.09; P = 0.07) compared with gilts fed the maintenance diet prior to mating. The post-mating diet had no effect on embryo survival. There were no treatment effects on blastocyst developmental stage, luteal surface area or progesterone release. Gilts receiving the high post-mating diet had heavier livers than those fed the maintenance post mating diet (1.45 v 1.08% of total bodyweight, SED = 0.07; P<0.001), suggesting that these gilts have a greater capacity to metabolize progesterone. Pre-mating nutritional status therefore appears to be a greater determinant of embryo numbers and survival than the post-mating diet. PMID- 10681005 TI - Reversed-phase liquid chromatography of proteins and peptides using multimodal copolymer-encapsulated silica. AB - Multimodal copolymer-encapsulated particles for liquid chromatography were prepared by bonding 1-octadecene and unsaturated carboxylic acids on silica particles (5 microm diameter, 300 A pores) for liquid chromatography of proteins. These multimodal copolymer-encapsulated particles can provide both hydrophobic and hydrogen bonding interactions with polar compounds. The chromatographic performance of these multimodal copolymer-encapsulated particles for peptide and protein separations was evaluated under reversed-phase conditions. Compared with typical C8-bonded silica, polymer-encapsulated particles were more stable in acidic mobile phases and provided better recoveries, especially for large proteins (Mr>0.5 x 10(6)). Totally hydrophobic polymer-encapsulated particles were found to produce broad peaks for proteins, and significant improvements were observed by introducing hydrophilic groups (-COOH) onto the polymer-encapsulated surface to form a multimodal phase. For the reversed-phase liquid chromatography of peptides and proteins, improved selectivity and increased solute retention were found using the multimodal polymer-encapsulated particles. More peaks were resolved for the separation of complex peptide mixtures such as protein digests using the multimodal polymer-encapsulated particles as compared to totally hydrophobic polymer-encapsulated particles. PMID- 10681006 TI - Enantiomeric separation of amino alcohols on protein phases using statistical experimental design. A comparative study. AB - Two LC supports often used for giving enantioselective retention were tested and compared in the reversed-phase mode using statistical experimental design. The two supports contain two different proteins, alpha1-acid glycoprotein or cellulase immobilised to silica particles, as the chiral selectors. The two chromatographic columns are commercially available as Chiral-AGP and Chiral-CBH. Twelve closely structurally related amino alcohols were used as the testing solutes. For each column three important mobile phase descriptors, that improve the chiral recognition, were chosen as variables and retention and separation factors were used as responses. All the tested solutes were separated using the two protein based supports. However, the highest enantioselectivities, i.e., separation factors higher than 10 were obtained using the Chiral-CBH column. The solute structure, e.g., distance between the nitrogen atom and the chiral carbon atom, and position as well as type of substituent in the aromatic ring highly influence the enantioselectivity on both columns. For one of the solutes the choice of mobile phase composition could be used to control the retention order of the two enantiomers. PMID- 10681007 TI - Analysis of pigmented high-molecular-mass grape phenolics using ion-pair, normal phase high-performance liquid chromatography. AB - A normal-phase LC method has been developed to analyze high-molecular-mass grape phenolic compounds. Samples are prepared by first isolating phenolics using C18 SPE. The analytical method uses a silica column and gradient elution with mobile phases of methylene chloride, methanol, formic acid and heptanesulfonic acid. This separation enables the analysis of these compounds from grape and wine samples in the presence of anthocyanins without extensive purification. Based on the elution order of proanthocyanidins and anthocyanins, phenolics elute in order of increasing molecular mass. Currently, it is not possible to identify all of the components separated in the chromatogram. PMID- 10681008 TI - Comparison of the performance of butanol and pentanol as modifiers in the micellar chromatographic determination of some phenethylamines. AB - A procedure was developed for the determination of several phenethylamines (amphetamine, arterenol, ephedrine, phenylephrine, phenylpropanolamine, mephentermine, methoxyphenamine, pseudoephedrine and tyramine), using micellar mobile phases of sodium dodecyl sulfate (SDS), a C18 column and UV detection. The drugs were eluted at short retention times with conventional acetonitrile-water or methanol-water mobile phases. In contrast, in the micellar system, they were strongly retained due to association with the surfactant adsorbed on the stationary phase, and needed the addition of butanol or pentanol to be eluted from the column. These modifiers allowed a simple way of controlling the retention. The chromatographic efficiencies obtained with the hybrid mobile phases of SDS-butanol and SDS-pentanol were also very high, mostly in the N=3000 7000 range, significantly greater than those achieved with a conventional acetonitrile-methanol-water mobile phase. Butanol and pentanol yielded similar selectivities, but the latter modifier permitted significantly shorter retention times than butanol, and was preferred to expedite the analysis of the pharmaceuticals. Most binary combinations of the nine phenethylamines can be resolved with these mobile phases. A mobile phase of 0.15 M SDS-5% pentanol was used to assay five of the phenethylamines (amphetamine, ephedrine, phenylephrine, phenylpropanolamine and pseudoephedrine) in 22 pharmaceutical preparations, which contained diverse accompanying compounds. The results agreed with the declared compositions and with those obtained with a mobile phase of methanol-acetonitrile 0.05 M phosphate buffer (pH 3) 10:5:85, with no interferences and relative errors usually below 2%. However, with the aqueous-organic mobile phase, the retention time for phenylephrine was too low and could not be usually evaluated. PMID- 10681009 TI - High-performance liquid chromatography of selenium compounds utilizing perfluorinated carboxylic acid ion-pairing agents and inductively coupled plasma and electrospray ionization mass spectrometric detection. AB - Increasing speciation demands in clinical chemistry, toxicology and nutrition have made the determination of the total elements in a sample inadequate; the amount of an element and the chemical forms in which it is present need to be known. Inductively coupled plasma mass spectrometry (ICP-MS) was used after high performance liquid chromatographic (HPLC) separation, as was electrospray ionization mass spectrometry (ESI-MS). The effect of variation of the number of carbon atoms in perfluorinated carboxylic acids used as ion-pairing agents for the separation of selenium compounds was examined. Trifluoroacetic acid (0.1%), pentafluoropropanoic acid (0.1%) or heptafluorobutanoic acid (0.1%; HFBA) were alternatively used as additives to methanol-water (1:99, v/v) solutions as mobile phases. Reversed-phase HPLC-ICP-MS with 0.1% HFBA in the mobile phase allowed more than 20 selenium compounds to be separated in 70 min in an isocratic elution mode; the separation of natural selenium-enriched sample extracts was examined and explained. The pH of the 0.1% HFBA solution was modified with hydrochloric acid or ammonia and the pH of the sample extracts before injection was modified in order to overcome unwanted double peak formation in the chromatograms of sample extracts. Oxidations of standard gamma-glutamyl-Se-methylselenocysteine and Se-methylselenocysteine were carried out using 30% H2O2 solution and identifications of selenium-containing oxidation products were made using HPLC ICP-MS and HPLC-ESI-MS. The principal organic oxidation product in both cases was methaneseleninic acid (MeSeO2H). PMID- 10681010 TI - Determination of halogenated mono-alcohols and diols in water by gas chromatography with electron-capture detection. AB - We have developed an analytical method for the detection of halogenated alcohols in water with particular focus on 3-chloro-1,2-propanediol and 3-bromo-1,2 propanediol. In this method the target analytes are extracted from water, derivatized with heptafluorobutyric anhydride, and then analyzed with gas chromatography with electron-capture detection. The effects of water, pH and seawater constituents on the method were investigated. Method detection limits for a 5 ml aqueous sample ranged from 0.14 microg l(-1) for 2-bromo-1,3 propanediol to 1.7 microg l(-1) for 1,3-dichloro-2-propanol (1,3DCP). PMID- 10681011 TI - Application of pressurized liquid extraction followed by gas chromatography-mass spectrometry to determine 4-nonylphenols in sediments. AB - A time- and solvent-saving method, pressurized liquid extraction (PLE), to extract 4-nonylphenol (4-NP) in sediment was developed. The effects of various operational parameters (i.e., temperature, pressure, etc.) for the quantitative extraction of 4-NP by PLE were investigated. The analytes were then identified and quantitated by a large-volume injection GC-MS technique. The 4-NP can be completely extracted by methanol at 100 degrees C and 100 atm combined with 15 min static and then 10 min dynamic extraction steps (1 atm = 101,325 Pa). Recovery of 4-NP in spiked blank kaolin samples was 98% with 5% RSD. The degrees of recovery of 4-NP in the spiked sediment samples from a reservoir and a polluted river were 111% with 4% RSD and 106% with 5% RSD, respectively. The perfect applicability of PLE for 4-NP was determined after testing it with spiked and aged samples. The extraction efficiency of the PLE was compared with conventional Soxhlet and bath ultrasonication extraction methods using the spiked sediment samples. PMID- 10681012 TI - Gas chromatographic profiling and screening for phenols as isobutoxycarbonyl derivatives in aqueous samples. AB - An efficient method is described for the simultaneous determination of phenol and 49 substituted phenols present in aqueous samples. The method is based on the extractive two-phase isobutoxycarbonyl (isoBOC) derivatization with subsequent solid-phase extraction (SPE) for the direct analysis by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Phenolic hydroxyl groups in acidic aqueous solutions were allowed to react with isobutyl chloroformate present in the dichloromethane phase containing triethylamine. The resulting isoBOC derivatives were then recovered by SPE using Chromosorb P in normal-phase partition mode, followed by direct GC and GC-MS analysis. Using this combined procedure, linear detector responses were obtained in the concentration range of 0.5-8 microg ml(-1), with correlation coefficients varying from 0.925 to 0.999 for most of the phenols studied except for 2,4-dinitorphenol (0.789). The temperature-programmed retention index (I) sets as measured on DB-5 and DB-17 dual-capillary columns of different polarity were characteristic of each isoBOC phenol derivative and thus, useful in the screening for isomeric phenols by I matching only. The mass spectral patterns, exhibiting characteristic [M-100]+, [M 200]+ and [M-300]+ ions for the mono-, di- and trihydroxybezenes, respectively with common ions at m/z 57, facilitated their rapid structural confirmation. The present method allowed rapid screening for phenols when applied to water samples spiked with phenols. PMID- 10681013 TI - Determination of reducing end sugar residues in oligo- and polysaccharides by gas liquid chromatography. AB - Reducing end sugar residues in maltodextrins and arabinoxylans are determined as alditol acetates by gas-liquid chromatography following reduction, acid hydrolysis and acetylation of the samples. After this conversion to alditol acetates, the reducing end sugars are thus separated from their acetylated aldose counterparts. The method allows to identify individual reducing end sugars quantitatively and is a good alternative for colorimetric reducing sugar assays and 1H-NMR analysis. To demonstrate the advantages of the method, an application in a study of enzymic solubilisation and degradation of water unextractable arabinoxylan from a flour squeegee fraction is described. PMID- 10681014 TI - Prediction of gas chromatographic retention indices of polychlorinated dibenzothiophenes on non-polar columns. AB - Polychlorodibenzothiophenes (PCDTs) have been found in several kinds of environmental samples. The lack of reference compounds has meant that very little is known about their gas chromatographic behavior. Here we discuss the retention of 19 authentic PCDTs and their sulfones on the widely used gas chromatographic stationary phases DB-5 and DB-5ms. The retention order is different from that of the polychlorodibenzofurans. The data generated allowed us to carry out a multiple linear regression to generate parameters for predicting the retention indices of unknown congeners based only on their structural features. PMID- 10681015 TI - One-step capillary isoelectric focusing for the separation of the recombinant human immunodeficiency virus envelope glycoprotein glycoforms. AB - One-step capillary isoelectric focusing was investigated as a rapid method to resolve the glycoforms of the heterogeneous recombinant human immunodeficiency virus (HIV) envelope glycoprotein (rgp 160sMN/LAI). The separation was performed in a poly(vinyl alcohol) (PVA) coated capillary using a mixture of ampholyte of narrow and wide pH range. A combination of saccaharose and 3-(cyclohexylamino)-1 propanesulfonic acid was shown to be the most efficient additive to avoid protein precipitation which occurs at a pH close to its pI. Although the calibration curve [isoelectric point (pI) vs. migration times] showed a non-linear relationship, an adequate linearity could be yielded for short pI ranges permitting to exhibit the acidic character of the different glycoforms of the rgp 160s MN/LAI (pI from 4.00 to 4.95). Reproducibility evaluated by comparing the performance of a polyacrylamide and a PVA coated capillary showed that low RSD values were obtained for intra-day (0.5 to 1.9%) and inter-day (1.6 to 7.6%) measurements using the PVA capillary. Moreover, the long term stability of the PVA capillary was demonstrated by measuring the variation of migration times of the protein markers for a long period of use. Finally, this method was able to differentiate the glycoform pattern of two close glycoproteins such as the rgp 160 of two sub-populations of the virus HIV-1. PMID- 10681016 TI - Determination of pesticides in waters by automatic on-line solid-phase extraction capillary electrophoresis. AB - The separation of seven pesticides by micellar electrokinetic capillary chromatography in spiked water samples is described, allowing the analysis of pesticides mixtures down to a concentration of 50 microg l(-1) in less than 13 min. Calibration, pre-concentration, elution and injection into the sample vial was carried out automatically by a continuous flow system (CFS) coupled to a capillary electrophoresis system via a programmable arm. The whole system was electronically coupled by a micro-processor and completely controlled by a computer. A C18 solid-phase mini-column was used for the pre-concentration, allowing a 12-fold enrichment (as an average value) of the pesticides from fortified water samples. Under the optimal extraction conditions, recoveries between 90 and 114% for most of the pesticides were obtained. PMID- 10681017 TI - Immunohistochemical study of S100-like protein in Eimeria brunetti and Eimeria acervulina. AB - We have investigated the expression of a calcium-binding protein, the S100 protein, in Eimeria brunetti and Eimeria acervulina stages. For this purpose, paraffin sections of distal ileum and bursa of Fabricius or duodenum from experimentally infected chickens were treated with anti-alpha-S100 (anti-alpha subunit of S100 protein) and anti-beta-S100 (anti-beta subunit of S100 protein) monoclonal antibodies and anti-S100 whole molecule polyclonal antibody. The avidin-biotin peroxidase method was used to demonstrate immunoreactivity. In the ileum, our results reveal a positive immunoreaction for the beta subunit and S100 whole molecule within the macrogametes of E. brunetti, whereas they were devoid of immunostaining after treatment of the paraffin sections with the anti-alpha S100 antiserum. Schizonts and oocysts of E. brunetti and all the E. acervulina stages gave a negative reaction after treatment with any of the three antiserum used in the study. This result indicated that the S100 protein molecules within these stages were not recognized by the antibodies, suggesting that these molecules are different from those identified in macrogametes of E. brunetti. By contrast, in the epithelial cells, lining the lumen of the bursa of Fabricius, macrogametes of E. brunetti were stained by the three antibodies used. These results may indicate the existence of metabolic adaptations that enable the parasite to invade tissue sites different from those where the parasite usually develops. PMID- 10681018 TI - Growth performance, meat quality and activities of glycolytic enzymes in the blood and muscle tissue of calves infected with Sarcocystis cruzi. AB - Growth performance and the pattern of glycolytic enzymes in the blood plasma were assessed during experimental Sarcocystis cruzi infection (1 x 10(5) sporocysts per calf) in six calves; five calves served as noninfected controls. At slaughter (68 or 88 days post infection), carcass weight, dressing percentages and several parameters of meat quality (pH, color brightness, rigor, water absorbing capacity, water binding capacity) were recorded. Moreover, enzyme activities were measured in muscle homogenates. Weight gain was significantly impaired by the infection. Activities of lactate dehydrogenase (LDH) and aldolase (ALD) significantly increased in the blood plasma of the infected calves during the chronic stage of the disease, while glucose-6-phosphate dehydrogenase (G6PDH) and isocitrate dehydrogenase (ICDH) were not significantly altered. This was accompanied by a significant decrease of enzyme activities in the Musculus longissimus dorsi (LDH, ALD), in the diaphragmatic musculature (ALD, G6PDH) and in the heart (LDH, ALD). Activities of LDH, ALD, ICDH and G6PDH were visualized by enzyme histochemistry within the developing sarcosporidial cysts. However, isoenzymes of parasite origin could not be demonstrated by agar-gel electrophoresis of muscle homogenates or blood plasma. It is concluded that sarcocystiosis of even moderate severity alters the performance of calves but not meat quality. Leakage of glycolytic enzymes from the affected muscles is the probable cause of increased plasma enzyme activities. Although these enzymes are also synthesized by the parasite, the contribution of parasite-derived enzymes to the observed changes of enzyme patterns remains in question. PMID- 10681019 TI - Cell-mediated immune response in calves to single-dose, trickle, and challenge infections with Fasciola hepatica. AB - A peripheral blood mononuclear cell (PBMC) proliferation assay was used to study the cell-mediated immune response in eight calves experimentally infected with Fasciola hepatica. Hypersensitivity-related eosinophil and mast-cell responses were also assessed. The primary infection of 500 metacercariae was administered either as a single-dose or as a trickle infection over a 4-week period. Calves were challenge-infected 4 months later with 100 metacercariae and slaughtered 24 weeks postprimary infection. Skin eosinophil counts (SEC) were determined prior to infection on the basis of the intradermal reaction (IDR) to phytohaemagglutinin (PHA). These counts correlated negatively with the mean fluke length but not with the fluke burden found at necropsy. At the end of the experiment, non-specific (PHA) and specific (excretory-secretory parasite, products, FhESAg, and whole-worm extract, FhSomAg) immediate type hypersensitivity IDR were elicited in contrast to delayed type hypersensitivity (DTH) responses. The SEC correlated with blood eosinophilia but not with parasite parameters. These findings suggest that the eosinophil response does not correlate clearly with the development of resistance to F. hepatica infection in cattle. A specific mononuclear cell response to FhSomAg was detectable as early as 7 days after infection in both infected groups, being significantly higher during the very early migratory phase of the juveniles in the single-dose infected calves than in the trickle infected calves. This response remained significantly higher in infected groups than in the control group throughout the experiment. Challenge elicited a significant proliferative response, less pronounced than after primary infection. No production of gamma-interferon (INF gamma) was recorded 3 weeks after challenge. At necropsy, the mean number of flukes recovered was similar in both infected groups, suggesting that the rate at which the infection is administrated has no effect on protective immunity. Hepatic lesions, similar in both infected groups, were characterised by marked eosinophil and mast-cell infiltration. Liver biopsies were performed and their diagnostic value is discussed. All results suggest that F. hepatica infection predominantly induces a Type-2 response in cattle, and that this response has little protective effect. PMID- 10681020 TI - Prevalence of hydatidosis among donkeys in northern Jordan. AB - One hundred and thirty donkeys (Equus asinus), aged between 5 months and 14 years of age, from the Irbid Governorate in northern Jordan were necropsied between November 1997-May 1999. Of these animals, 16.9% had hydatid cysts in either their lungs and/or livers. No donkeys of 3 years of age or less were infected, where as 33.3% (22 of 66) aged 4 years or greater were infected. Intensity of infection increased with age in a linear fashion. The prevalence also increased with age approaching an asymptotic prevalence of 1 in the oldest animals. This implied there was minimal regulation of the parasite population by intermediate host immunity. The numbers of cysts in the donkeys were increasing at a rate of 0.48 cysts per year from 0.054 infections. The frequency distribution was highly aggregated, consistent with a negative binomial distribution indicating infection of donkeys was not random. PMID- 10681021 TI - Sero-epidemiological study of Taenia saginata cysticercosis in Belgian cattle. AB - A sero-epidemiological survey of Taenia saginata cysticercosis was carried out to determine the prevalence of the infection in cattle presented for slaughter in Belgium. Between November 1997 and June 1998, a total of 1164 serum samples were collected in 20 export abattoirs. Meat inspection was routinely carried out by veterinary inspectors. Serum samples were examined for circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked-immunosorbent assay (Ag-ELISA). Thirty six serum samples (3.09%) were found positive in the Ag ELISA, while by meat inspection on the same animals cysticerci were detected in only three carcasses (0.26%). Sero-prevalence was positively correlated with the age of the animals. The sero-prevalence found in this study was more than 10 times higher than the annual prevalence (0.26%) reported by the Institute for Veterinary Inspection. This study clearly indicates that the classical meat inspection techniques detect only a minor fraction of the carcasses infected with cysticerci. PMID- 10681022 TI - Seasonal development and survival of equine cyathostome larvae on pasture in south Louisiana. AB - Cyathostome development and survival on pasture in subtropical climates of the US have yet to be completely defined and available data on seasonal transmission are minimal. In an attempt to study this phenomenon, a group of pony mares and their foals was maintained on a naturally contaminated pasture in southern Louisiana. Fecal egg counts (FEC) and numbers of infective third stage larvae (L3) kg(-1) dry herbage were recorded biweekly during two time periods, from January 1986 through December 1988, and September 1996 through October 1997. A FEC rise occurred during the late summer-early autumn which preceded the peak of L3 on pasture during the winter season. The numbers of cyathostome L3 were reduced during the hottest months of the year due mainly to daily minimum temperatures above 18 degrees C, and in winter during short freezing spells when daily minimum temperatures dropped below 0 degrees C. Tilling of the pasture reduced the number of cyathostome L3 during the early winter months but this is an efficacious measure only if horses are given an effective anthelmintic treatment prior to being returned to pasture. The data collected suggest that parasite reduction in southern Louisiana is possible using a treatment program with treatment beginning at the end of September and continuing through the end of March. PMID- 10681023 TI - The effect of oxfendazole terminated infections with Haemonchus contortus on the development of immunity in sheep. AB - The relative contribution of third (L3), fourth (L4) or adult stages of Haemonchus contortus to the development of immunity was evaluated in three groups of sheep subjected to infections terminated by oxfendazole treatments at the L3, L4 or adult stage. A control group did not receive immunising infections. All the groups were challenged with 5000 L3, to evaluate the protection provided by the different protocols. All sheep were necropsied at the end of the experiment to count the abomasal worm burdens. A marked reduction in egg counts after challenge infection was only observed in sheep in which the infection was terminated in the adult stage (Group 4). A significant reduction in worm burden was also observed in Group 4. The immunising infections and/or the challenge infection resulted in moderately elevated IgG antibody levels against L3, L4 and adult somatic antigens in all the groups. In contrast, a strong IgG response against H. contortus excretory/secretory (ES) antigens was observed in the groups in which the immunising infection was terminated in the L4 and the adult stage. An elevated lymphocyte proliferation response against Haemonchus ES antigens was found only in the group that had their immunising infection terminated at the adult stage. The combined data suggest that exposure to and elicited immunological responses to ES antigens are important for the development of immunity against H. contortus. PMID- 10681024 TI - Detection of Oestrus ovis and associated risk factors in sheep from the central region of Yucatan, Mexico. AB - A cross-sectional epidemiologic study was conducted in order to detect the presence of and to estimate the seroprevalence of Oestrus ovis L. infection in flocks of sheep from the central region of the state of Yucatan, Mexico. The risk factors associated with disease were also identified. A sample size of 10 animals per farm was used to detect seropositive animals, considering a 30% prevalence and 95% confidence level. Blood samples of 689 sheep from 88 flocks were collected and a questionnaire with questions about the flock and the host was applied. The thin layer immune assay test was used. The risk factors were screened using logistic regression procedures. 77% of the flocks had at least one positive animal with antibodies against O. ovis. The overall seroprevalence and standard error was 30.6 +/- 3.5%. Only flock size and sheep nose color showed association (P < 0.05) with the disease. The odds ratios for flocks with less than 11 and with 11 to 25 sheep, as related to herds with 25 or more sheep, were 0.74 and 1.73, respectively. Sheep with dark noses had a higher risk (OR = 1.46) compared with sheep having light noses (P < 0.05). PMID- 10681025 TI - Possible risk factors on Queensland dairy farms for acaricide resistance in cattle tick (Boophilus microplus). AB - A case control study was carried out within a cross-sectional survey designed to investigate the management by Queensland dairy farmers of the cattle tick Boophilus microplus. Although 199 farmers were surveyed, data on acaricide resistance were only obtained from 66 farms. Multiple models were used to predict the probability of acaricide resistance associated with 30 putative risk factors. The region of the state in which the farm was located and the frequency of acaricide application were consistently associated with acaricide resistance. The risk of resistance to all synthetic pyrethroids (Parkhurst strain) was highest in Central Queensland and increased when more than five applications of acaricide were made in the previous year, when spray races were used and when buffalo fly treatments with a synthetic pyrethroid were applied frequently. The probability of resistance to amitraz (Ulam strain) was highest in Central Queensland, increased when more than five applications of acaricide were made in the previous year, and decreased on farms when a hand-spray apparatus was used to apply acaricides to cattle. The probability of resistance to flumethrin (Lamington strain) was highest in the Wide Bay-Burnett region. PMID- 10681026 TI - Field observations of the host-parasite relationship associated with the common horse bot fly, Gasterophilus intestinalis. AB - Behavior of Gasterophilus intestinalis (Diptera: Gasterophilidae) was observed in the laboratory and field over a period of 10 years. Mating occurred in a frenzied manner as flies emerged from pupae. Males attempted to copulate with females that had not fully emerged from the pupal case. Mating was never observed in the field or near the horse. Observations suggest that fecal piles of the horse are used as the mating site for the newly hatched flies. Activation of host-seeking behavior was found to occur early and remain throughout the life span of the adult as it sought multiple hosts for oviposition. Evidence is presented supporting the use of olfaction and vision in finding the host. Interest by flies in shadows and linear high-contrast objects was observed. Methods used by the flies to maintain close proximity with the host are discussed including flight with the host and avoidance of host defenses. Recapture methods were successful in determining the time taken to reestablish at the host, and extent of travel to the host. The presence of kairomone(s) within horse secretions is suspected. PMID- 10681027 TI - WAAVP/Pfizer award for excellence in veterinary parasitology research. My involvement in, and some thoughts for livestock parasitological research in Australia. AB - Being presented with the WAAVP Pfizer award for excellence in parasitological research is the pinnacle of my career. In accepting I acknowledge the support that I have received from workmates, colleagues, friends and family over the years that I have been involved in this field of endeavour. Parasitic disease is the most significant threat to the Australian sheep industry. A lack of understanding of drug action, the absence of epidemiologically-based treatment programs and incorrect or excessive chemical use has resulted in the development of worm, lice and blowfly parasites which are resistant to most existing chemotherapeutic compounds. During the past decade, difficulties in sustainable control of parasitic disease, the decline in demand for wool products and competition from less expensive synthetic fibre has halved the sheep population and profitability of the industry. Notwithstanding this, a 'right-sized', sustainable industry is emerging which will require effective chemotherapy to be the cornerstone of parasite control. Chemical intervention in parasitic disease is therefore here to stay but the paucity of new antiparasitic products in the short term dictates that present therapeutics are all that producers will have for the foreseeable future. This situation will necessitate innovative practices and formulations to provide more cost effective, efficient drug performance and to extend parasiticide life. However, the development of multiple drug resistance and reduction in funds for parasitological research seriously compromises our ability to confront these demands. With the patent life of all but the most recent macrocyclic lactone (ML) compounds lapsing, low cost development of bioequivalent generic formulations and options for innovative strategies to increase performance and market share are eagerly sought. The key to efficient drug use lies in a detailed understanding of the pharmacokinetic principles of drug action and the host animal's physiological responses to identify procedures which maximise drug availability--in essence giving the drug the best chance to work. It is therefore evident that the how, where and why of drug exchange between the bloodstream and the gastrointestinal tract are of such interest. Of particular importance is identification of the kinetic and dynamic behaviour of drug in the gastrointestinal tract (GIT) and the role of biliary secretion and metabolic fate of biliary (and non-biliary) compounds. The extent to which biliary secreted parent drug and/or metabolites are presented to the gut lumen, reabsorbed as free compound or after deconjugation by large bowel bacteria, and participate in the enterohepatic cycle is a major contributor to parasite exposure. Integrating parasiticide disposition with host physiology, particularly relating to such aspects as gastrointestinal function, feed intake and body condition has demonstrated the value of 'whole body' pharmacokinetic studies to identify processes to increase drug efficacy. Novel formulation modifications to provide 'targeted' drug delivery including carrier technology, sustained release devices and site-directed formulations can manipulate pharmacokinetic disposition to direct or extend drug availability to the parasite infection. These research directions should be undertaken as collaborative projects between research organisations and the veterinary pharmaceutical industry. With the identification of methods to improve drug action, emphasis must then be directed towards effective communication for their implementation. It will be vital that parasitologists move out of the laboratory to actively disseminate knowledge to the producer. We are ambassadors for our profession and if we fail to communicate well the perception, and indeed the value, of our work can be at risk. PMID- 10681028 TI - WAAVP/Pfizer award for excellence in teaching veterinary parasitology: teaching of veterinary parasitology--quo vadis? AB - Some thoughts on training and recruitment of academic teachers and future trends in teaching veterinary parasitology are presented with emphasis on the European situation. It is underlined that research is an indispensable basis for academic teaching. Besides a broad scientific background of the teacher, motivation and teaching methods are also important. Many academic teachers do not receive formal training in teaching methods. In order to improve future education, training of staff members in teaching methods should be promoted. Quality control of teaching and research, already established in many schools, should generally be introduced. Teaching is mostly underestimated in relation to research. Therefore, more weight should be placed on the former both in selecting scientists for the career as academic teachers and in evaluating and ranking departments for their academic activities. In the future veterinary medicine will have to cope with profound changes in the society and the veterinary profession, and the progressing European unification will enhance trends for internationalizing teaching curricula. Therefore, veterinary medicine has to reconsider the teaching subjects and methods and to lay more emphasis on flexibility, skills of problem solving and self-learning and on training for life-long learning. At present there is an ongoing discussion on the question how to teach veterinary medicine, including veterinary parasitology. There are various options, and some of them are discussed, namely, the disciplinary and the problem-based/organ-focussed approaches. It is concluded that for teaching of veterinary parasitology and related disciplines a combined disciplinary and problem-based approach offers the best chances for fulfilling the requirements of teaching for the future. In the curriculum of undergraduate teaching of veterinary medicine at least 70-90 h should be dedicated to veterinary parasitology using a disciplinary and taxonomic approach. Additional hours are required for instructions on clinical cases in approaches focussed on animal species and/or organ diseases. As there is a need for discussing teaching issues, post-graduate specialization, and continuing education in parasitology and related disciplines on national and international levels, it is recommended to WAAVP to include regular workshops on teaching in the programmes of the biannual conferences, and to establish a permanent committee which should collect information and submit proposals for improvement of teaching veterinary parasitology. PMID- 10681029 TI - Performance of the ELISA test for swine cysticercosis using antigens of Taenia solium and Taenia crassiceps cysticerci. AB - Studies were conducted to evaluate antigens of Taenia solium (Tso) and Taenia crassiceps (Tcra) cysticerci in the ELISA test for the diagnosis of swine cysticercosis. The samples analyzed were cysticercosis positive and negative control sera and heterologous sera. Four antigens were assayed: vesicular fluid (VF) and crude (T) Tcra and scolex (S) and crude (T) Tso. All antigens showed good performance, but VF-Tcra was the best followed by T-Tcra. Sensitivity rates of ELISA were respectively, in 2nd and 3rd standard deviation cut-offs, 96.0 and 80.0% for the VF antigen and specificity of 97.5 and 100.0%. Cross-reactivity was verified only for hidatidosis and ascaridiosis. Due to the high performance observed, the ELISA test using Tcra antigens should be recommended for the diagnosis of swine cysticercosis. PMID- 10681030 TI - Canine echinococcosis: an alternative for surveillance epidemiology. AB - The essential activities for programmes of cystic echinococcosis control are the census of all dogs from the program and identification of parasitised animals. Currently, in South America evaluations and epidemiological surveillance are based on the administration of arecoline hydrobromide. This method has the disadvantage of increasing environmental pollution and risk for operators and owners of treated dogs. A genus-specific ELISA capture method has been employed for recently issued faeces and the confirmation of positive examination was performed by dog autopsies. Our work presents an alternative method based on collection of dry field-dispersed faeces, followed by serological diagnosis by Copro-ELISA and confirmation by Copro-Western blot. If Copro-ELISA were used to define positive samples of dry faeces, the Copro-Western blot assay would provide 70% sensitivity and 100% specificity. Global efficiency of the system using dry faeces would reach 76%, allowing epidemiological surveillance to be oriented to analysis of surface units instead of dog as measurement unit. PMID- 10681031 TI - Cooperia pectinata and C. punctata, parasites of the abomasum of cattle in northern Cameroon (Central Africa). AB - Cooperia pectinata Ransom, 1907 and C. punctata von Linstow, 1907 are common trichostrongyles of zebu cattle in Africa. Their intestinal localization within the digestive tract is considered by many authors to be exclusive. Nevertheless, some limited surveys in Malagasy, Mauritania, The Gambia and Cameroon reported the presence of both Cooperia species in the abomasum. The present survey was carried out in a slaughterhouse of northern Cameroon on 17 zebu cattle and confirms the infection of the small intestine and the abomasum by the two species within the total number of cattle examined. Abomasal infections especially with Cooperia punctata were heavier than those in the intestine. Due to the movements of herdbreeders in Central Africa, and to the preliminary results obtained in Mauritania, The Gambia, Burkina Faso and Malagasy, abomasal localization of C. pectinata and C. punctata may be encountered in very large areas of Africa, and that cooperiosis may contribute together with Haemonchus species to the digestive disorders involving the abomasum. PMID- 10681032 TI - The efficacy of levamisole, and a mixture of oxfendazole and levamisole, against the arrested stages of benzimidazole-resistant Haemonchus contortus and Ostertagia circumcincta in sheep. AB - Sheep were allowed to graze pasture that had been seeded with benzimidazole resistant Haemonchus contortus and Ostertagia circumcincta in order to acquire a burden of arrested larvae. Following housing, sheep were dosed orally with either oxfendazole at a dose rate of 4.7 mg/kg (to confirm the benzimidazole-resistant status of the species of nematode), levamisole at a dose rate of 7.5 mg/kg, or an oxfendazole/levamisole mixture at a dose rate of 4.6 mg/kg oxfendazole and 8.1 mg/kg levamisole. The efficacies of the treatments were assessed by estimation of the arrested larval burden in the abomasum of each sheep, either at 10 or 11 days (oxfendazole and oxfendazole/levamisole mixture), or 12 or 13 days (levamisole), after treatment. Compared to the untreated controls, the protection afforded by a single dose of either levamisole or the oxfendazole/levamisole mixture was >99% against the arrested stages of both Haemonchus contortus and Ostertagia circumcincta. Treatment with oxfendazole confirmed the benzimidazole-resistance status of the two species. PMID- 10681033 TI - Hypolipemia associated with the wasting condition of rabbits infected with Strongyloides papillosus. AB - Rabbits develop a wasting condition in the intestinal stage of Strongyloides papillosus infection. Serum inflammatory cytokine and lipid profiles were investigated in five rabbits infected with S. papillosus and five uninfected pair fed controls to ascertain whether the disease is inflammatory cytokine-mediated cachexia. Tumor necrosis factor alpha (TNF alpha) was detected in one infected animal at Day 7 after infection. Interleukin (IL)-1 was detected in three infected, and one control, animals at Day 28. IL-6 remained unchanged in both the groups. Infected animals developed hypolipemia, including hypotriglyceridemia in the intestinal stage of infection. Control animals lost body weight in the same manner as the infected animals, but had elevated cholesterols and phospholipids with normal triglyceride concentrations. The results suggested that the wasting condition has no association with cachexia induced by TNF alpha. IL-1 or IL-6, and that hepatic function for lipid synthesis is affected during the intestinal stage of S. papillosus infection. PMID- 10681034 TI - Variability of resistance in goats infected with Haemonchus contortus in Brazil. AB - The variability between and within breeds with respect to nematode egg counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) was studied in 36 yearling female goats of the Caninde (15), Bhuj (6) and Anglo-Nubian (15) breeds, exposed to Haemonchus contortus. Nematode-free goats were turned to a contaminated paddock in late February. From then on, fecal egg per gram counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) were determined at 2-week intervals up to Week 18. The EPG, transformed as [log(EPG + 75)], varied (P < 0.01) between goats within breeds and between weeks of exposure, but not between goat breeds (P > 0.05). PCV and Hb were affected by goat breeds (P < 0.05), by goats within breeds (P < 0.01) and by weeks of exposure (P < 0.01). Anglo-Nubians had higher (P < 0.01) PCV and Hb than Caninde; Bhuj had intermediate values. There were two EPG rises; one between Weeks 6 and 10 and the other between Weeks 14 and 16. The within breed variability was marked during the EPG rise on Week 6, when individual egg counts ranged from 130 to 2500. The EPG rises coincided with drops in Hb. PCV presented a similar trend, though not as marked. Haemonchus was responsible for more than 95% of nematode eggs counted. Considering the goat as experimental unit, the correlation coefficients (r) were: -0.45, P = 0.0064, between log(EPG + 75) and PCV; and -0.53, P = 0.0009, between log(EPG + 75) and Hb. The negative correlation between egg counts and blood values suggested breed differences in PCV and Hb were related to resistance to H. contortus infection and/or to its effects. PMID- 10681035 TI - Use of detomidine hydrochloride as an adjunct for studying first-stage Gasterophilus intestinalis (Diptera: Gasterophilidae) in the tongue of the horse. AB - A synthetic alpha-2 adrenergic agonist, detomidine hydrochloride, was used in the study of in vivo activity of Gasterophilus intestinalis (Diptera: Gasterophilidae) during migration in the tongue of the horse. Use of the drug allowed the investigator to manipulate the tongue and closely observe the movement patterns and tissue disturbance caused by burrowing first-stage larvae. Detomidine hydrochloride should be utilized in studies of drug efficacy and larval biology, whenever possible, to avoid the need to sacrifice the horse. PMID- 10681036 TI - Two-step purification method of vitellogenin from three teleost fish species: rainbow trout (Oncorhynchus mykiss), gudgeon (Gobio gobio) and chub (Leuciscus cephalus). AB - A two-step purification protocol was developed to purify rainbow trout (Oncorhynchus mykiss) vitellogenin (Vtg) and was successfully applied to Vtg of chub (Leuciscus cephalus) and gudgeon (Gobio gobio). Capture and intermediate purification were performed by anion-exchange chromatography on a Resource Q column and a polishing step was performed by gel permeation chromatography on Superdex 200 column. This method is a rapid two-step purification procedure that gave a pure solution of Vtg as assessed by silver staining electrophoresis and immunochemical characterisation. PMID- 10681037 TI - Two-step fast protein liquid chromatographic purification of the Serratia marcescens hemolysin and peptide mapping with mass spectrometry. AB - The pore forming toxin of Serratia marcescens (ShlA) is secreted and activated by an outer membrane protein (ShlB). Activation of inactive ShlA (termed ShlA*) by ShlB is dependent on phosphatidylethanolamine (PE). Activation may be a covalent modification of ShlA. To test this hypothesis, the responsible activation domain (in the N-terminal 255 amino acids of ShlA) was isolated from whole bacteria with 8 M urea in an inactive form (ShlA-255*) and from the culture supernatant in an active form (ShlA-255), followed by a two-step purification by anion-exchange chromatography and gel permeation chromatography. Comparison of a tryptic peptide map of both forms with subsequent electrospray mass spectrometry (ES-MS) and sequencing by tandem ES-MS revealed no modification. These data imply that ShlB presumably imposes a conformation on ShlA-255 that triggers activity. PMID- 10681038 TI - Preparative two-step purification of recombinant human basic fibroblast growth factor from high-cell-density cultivation of Escherichia coli. AB - Aggregation and precipitation are major pitfalls during bioprocessing and purification of recombinant human basic fibroblast growth factor (rh-bFGF). In order to gain high yields of the soluble protein monomer with high biological activity, an efficient downstream process was developed, focussing on the combination of expanded bed adsorption (EBA) and heparin chromatography. After expression in E. coli TG1:plambdaFGFB, cells were harvested and washed; then the rh-bFGF was released via high pressure homogenization. The high viscosity of the feedstock of about 40 mPa s, showing non-newtonian behaviour, was reduced to 2 mPa s by the addition of DNase. The homogenate (5.6 l) was loaded directly on an expanded bed column (C-50) packed with the strong cation-exchanger Streamline SP. In the eluates, histone-like (HU) protein was identified as the main protein contaminant by sequence analysis. The thermodynamics and kinetics of rh-bFGF adsorption from the whole broth protein mixture were determined in view of competition and displacement effects with host-derived proteins. Optimal binding and elution conditions were developed with knowledge of the dependence of rh-bFGF adsorption isotherms on the salt concentration to allow direct application of eluates onto Heparin HyperD. This affinity support maintained selectivity and efficiency under CIP and over a wide range of flow-rates; both is advantageous for the flexibility of the purification protocol in view of a scalable process. Remaining DNA and HU protein were separated by Heparin HyperD. The endotoxin level decreased from approximately 1,000,000 EU/ml in the whole broth to 10 EU in 3 mg bFGF per ml. The final purification protocol yields >99% pure rh-bFGF as judged from SDS-PAGE and MALDI-TOF mass spectrometry with high mitogenic activity (ED50=1-1.5 ng/ml) of the lyophilized sample. In comparison to the conventional process, the overall protein recovery rose by 15% to 65% with saving time and costs. PMID- 10681039 TI - Purification of the Ca2+-binding protein S100A1 from myocardium and recombinant Escherichia coli. AB - S100A1 is a new regulatory protein of myocardial contractility that is differentially expressed in early and late stages of myocardial hypertrophy. In order to further investigate the multiple functions of S100A1 in various assay systems we developed a new strategy for isolating biologically active S100A1 protein. After EDTA extraction of myocardium or recombinant bacteria, S100A1 was purified by Octyl-Sepharose hydrophobic interaction chromatography and HiTrapQ anion-exchange chromatography yielding 1.4-2.0 mg/100 g wet tissue and 0.7-1.0 mg/100 ml bacterial culture. Native porcine as well as human recombinant S100A1 revealed biological activity in physiological and biochemical assays. PMID- 10681040 TI - Multi-step purification strategy for RANTES wild-type and mutated analogues expressed in a baculovirus system. AB - RANTES (regulated on activation, normal T cell expressed and secreted), a C-C chemokine, is one of the major HIV-suppressive factors produced by CD8+ T cells. Wild-type RANTES and genetically modified analogues were expressed in a baculovirus system and purified from cell culture supernatants employing a multi step strategy based on affinity and RP-HPLC. Quantification and purity control of the final proteins were carried out by capillary electrophoresis using the synthetic or the recombinant wild-type RANTES as a reference. The procedure here reported requires only three days to obtain 0.016-0.270 mg of the pure and characterised proteins, starting from 370-900 ml of culture media, and is suitable for the analysis of a large number of RANTES analogues. PMID- 10681041 TI - Purification of the c-erbB2/neu membrane-spanning segment: a hydrophobic challenge. AB - High quality purification of membrane-spanning peptides and proteins remains a challenging problem. In this work we describe a tailored chromatographic purification of a synthetic 35-residue peptide corresponding to the transmembrane region of the tyrosine kinase receptor c-erb2/neu. Composed to over 70% by the amino acids alanine, isoleucine, leucine, phenylalanine and valine, this peptide presents a very hydrophobic character. Product isolation from the complex peptide mixture, obtained after acid cleavage of the resin matrix used during the solid phase synthesis, represents a difficult task. We propose a three step strategy based on gel permeation and reversed-phase high-performance liquid chromatography, using aprotic polar solvent mixtures. The challenge consisted in obtaining a sufficient amount of an extremely pure sample, in order to allow structural analysis by NMR spectroscopy. Keeping trace of the synthetic peptide throughout the different purification steps was assured by MALDI TOF mass spectrometry, and the final product purity was checked by coupled liquid chromatography-ESI TOF mass spectrometry. PMID- 10681042 TI - Chromatographic purification of the C(H)2 domain of the monoclonal antibody MAK33. AB - The C(H)2 domain, one of the constant domains of the murine monoclonal antibody MAK33 (immunoglobulin subtype K/IgG1) was expressed in Escherichia coli forming insoluble inclusion bodies (IBs) and purified by a three-step process including a denaturation-renaturation step, hydrophobic interaction and gel permeation chromatography. After disrupting the cells, the soluble protein fraction was removed by several centrifugation steps. The isolation of the IBs from the cell fragments was achieved by solubilizing the IBs with 6 M guanidinium hydrochloride (GdmCl) and 0.1 M 1,4-dithioerythrit (DTE) to reduce all disulfide bonds. After refolding the C(H)2 domain, 1.5 M (NH4)2SO4 was added to the protein solution in order to precipitate contaminations. Then the protein was loaded on a Butyl Sepharose fast flow column and eluted with a linear gradient [1.5-0 M (NH4)2SO4]. As the last purification step a gel permeation chromatography was run on a Superdex 75 prep grade. Finally, the purity of the C(H)2 protein was determined by a silver-stained sodium dodecyl sulfate polyacrylamide gel. We achieved a typical yield of 0.5 mg pure protein per 1 g of wet cells. PMID- 10681043 TI - Three-step purification of a fragment of the large immunophilin FKBP52. AB - PPIases catalyze the interconversion of cis and trans isomers of peptidyl-prolyl (Xaa-Pro) bonds in peptide and protein substrates. The PPIase family comprises three subfamilies, two of which interact with immunosuppressant drugs and are therefore termed immunophilins. One subgroup of the immunophilins are the FK506 binding proteins (FKBPs). FKBPs of a relative molecular mass higher than 40 000 also display chaperone activity and are part of the multichaperone complex that Hsp90 forms with substrate proteins. Their function in this chaperone complex is still enigmatic. To further characterize the function of FKBP52 we want to analyze constructs of FKBP52-fragments. Here we describe a fast and effective three-step purification procedure for a fragment of FKBP52 with a relative molecular mass of 48000, termed FKBP52-123, consisting of affinity chromatography, anion-exchange column and gel-permeation chromatography. A yield of 1 mg pure protein per gram of cells was achieved. PMID- 10681044 TI - Purification of a viral coat protein by an engineered polyionic sequence. AB - Virus-like particles composed of the polyoma coat protein VP1 were produced as a central building block of an artificial vector system for gene therapy. For this purpose, recombinant VP1 was expressed in E. coli. Classical purification schemes resulted only in low yields of protein. Therefore, we developed a new affinity purification procedure. We decided to use a polyionic sequence containing eight glutamic acid residues which allows efficient purification using ion-exchange chromatography. This peptide was inserted in a solvent exposed loop on the surface of VP1. After recombinant expression and cell lysis the first purification and concentration step consisted of a fractionated ammonium sulfate precipitation. The resuspended VP1 was loaded on an anion-exchange column. Elution with ca. 600 mM NaCl yielded almost homogeneous protein. Subsequently a size exclusion chromatography was performed to separate the pentameric VP1 from higher oligomeric and aggregated material. In contrast to wildtype VP1 the highly charged mutant form showed no significant tendency to aggregate. To demonstrate the functional state of the VP1 mutant, the in vitro assembly was investigated. At conditions similar to those for wildtype VP1 assembly, the mutant protein could form homogeneous virus-like particles. PMID- 10681045 TI - Expression, purification and biological properties of the carboxyl half part of the HTLV-I surface envelope glycoprotein. AB - The carboxyl half of the surface envelope protein of HTLV-I contains the major immunodominant and neutralizable domains. Using two affinity chromatography steps and a combination of high salt concentration and non-ionic detergent, we purified this part of the envelope protein from Escherichia coli. Analysis of some immmunological and biological properties of this protein indicated that it was folded in a way that preserved the correct structure of this domain of the HTLV-I envelope protein. It could be utilized in structural studies to further understand the mechanisms of HTLV-I entry and to better define the component(s) of an effective vaccine. PMID- 10681046 TI - Purification of respiratory syncytial virus F and G proteins. AB - Respiratory syncytial virus (RSV) is the most important cause of severe lower respiratory tract infections of infants in industrial nations. In addition, the participation of RSV in the genesis of asthma is under discussion. The RSV glycoproteins F and G have key positions in the viral pathogenesis. At present no satisfactory protein purification protocols are available for these proteins. The methods published for the G protein using preparative SDS-PAGE or immunoaffinity chromatography yield only small amounts of purified G protein that has partially lost its antigenicity. We describe a three-step purification protocol for these glycoproteins. RSV-infected HEp-2 cells were lysed by a Triton X-100 containing buffer. The viral proteins were captured by QAE-Sephadex A-50 material in a batch procedure. A first elution with 100 mM NaCl led to a crude F protein fraction, and a second elution with 300 mM NaCl led to a crude G protein fraction. The F protein was further purified on a Lentil-lectin Sepharose 4B column and finally polished using a Resource Isopropyl column. Lentil-lectin Sepharose 4B was also used to purify the G protein from the crude fraction, but polishing of the G protein was carried out on a Resource Q column. Homogenous RSV-F and RSV-G proteins were obtained by this protein purification protocol. No loss of antigenicity could be observed during this procedure as the highly purified viral proteins remain detectable by a set of monoclonal antibodies and specific antisera. The G protein was isolated as a 90000 monomer, whereas the purified F protein was recovered as a functional homodimer of 140000. PMID- 10681047 TI - Three-step purification of bacterially expressed human single-chain Fv antibodies for clinical applications. AB - We have obtained a cell line which secretes a human monoclonal IgM (B7) reacting with the myosin heavy chain of human heart. We have constructed single-chain fragments (scFv) of B7. The scFv may be useful for the imaging of myocardial necrosis after myocarditis, cardiac drug toxicosis or graft rejection. The aim of our work was to purify the scFv for immunoscintigraphy. We describe several purification steps including immobilized metal affinity chromatography (IMAC), anti-c-myc monoclonal antibody affinity chromatography, size-exclusion chromatography with Superdex 75 HR 10/30 and ion-exchange chromatography (mini Q TM 30Q). PMID- 10681048 TI - Purification of the membrane binding domain of cytochrome b5 by immobilised nickel chelate chromatography. AB - The purification of a eukaryotic membrane protein has been achieved using a prokaryotic expression system. Bovine cytochrome b5 is an integral membrane protein (Mr approximately 16500). It comprises of a globular haem containing catalytic domain positioned at the N-terminus of the protein and a hydrophobic membrane binding segment at the C-terminus. The membrane binding domain (MBD) is resistant to purification using conventional strategies that have proved successful in isolating the soluble haem containing fragment. We report here a versatile purification method for the isolation of the MBD involving a gene fusion system. The fusion protein incorporates thioredoxin at the amino terminus and six histidines as the metal affinity binding site followed by cytochrome b5 in a pET expression system. This supports high level expression of cytochrome b5 in E. coli C43(DE3) cells. The fusion protein is effectively solubilised from lysed cells with Triton X-100. A step gradient elution with imidazole under non denaturing conditions on a His-Bind nickel chelate affinity column, saturated with proteins as a crude cell extract, purified the protein in a single step. Proteolytic digestion of pure fusion protein, with trypsin, yielded the MBD. This fragment was further purified by RP-HPLC to a final yield of approximately 10 mg/l. PMID- 10681049 TI - Novel, rapid purification of the membrane protein photosystem I by high performance liquid chromatography on porous materials. AB - New porous materials have been tested for their potential to speed up purification of membrane proteins. As an example the purification of photosystem I, a light-driven electron pump from the cyanobacterium Synechocystis PCC6803, was optimized. The combination of two HPLC steps (an anion-exchange chromatography followed by a hydrophobic interaction chromatography) yields homogeneous monomeric or trimeric photosystem I as determined by gel filtration and gel electrophoresis. In comparison to traditional purification schemes our method is at least three-times faster and allows for easy scale-up. PMID- 10681050 TI - Three-step chromatographic purification procedure for the production of a his-tag recombinant kinesin overexpressed in E. coli. AB - A kinesin gene has been cloned by RT-PCR (reverse transcription polymerase chain reaction) from Trypanosoma brucei and the corresponding protein overexpressed as a recombinant His-tag (histidine-tag) kinesin in E. coli in order to study its biochemical properties and to determine its three-dimensional structure by X-ray crystallography. Starting from several liters of culture, an ultrasonic homogenizer was used for cell disruption and an unclarified feedstock was obtained. From this homogenate, a protein was then purified by immobilized metal affinity chromatography (IMAC) using expanded bed adsorption (EBA) technology (Streamline chelating). For this capture step, 100% of the recombinant protein was purified with more than 90% of purity. This step was followed by ion-exchange chromatography (Q Sepharose Fast Flow) for intermediate purification (96% purity, 53% recovery) and by size-exclusion chromatography with Superdex 75 as a polishing step (99% purity, 93% recovery). We then separated two forms of kinesin, a dimer (70%) and a monomer (30%). It was then possible to purify His tag recombinant protein directly from feedstock in a rapid and efficient way and to isolate two forms of kinesin. PMID- 10681051 TI - Improved purification of the membrane-bound hydrogenase-sulfur-reductase complex from thermophilic archaea using epsilon-aminocaproic acid-containing chromatography buffers. AB - A hydrogenase-sulfur reductase (SR) complex was purified from membrane preparations of the extremely thermophilic, acidophilic archaeon Acidianus ambivalens using a combination of sucrose density gradient centrifugation and column chromatography (FPLC). All chromatographic steps were performed in the presence of 0.5% epsilon-aminocaproic acid resulting in the elution of the SR complex as a sharp peak. In contrast, chromatography using buffers without epsilon-aminocaproic acid, or in the presence of detergents, were not successful. The purified A. ambivalens SR complex consisted of at least four subunits with relative molecular masses of 110000, 66000, 39000 and 29000, respectively. A similar procedure was applied to purify the membrane-bound hydrogenase from Thermoproteus neutrophilus, a non-related extremely thermophilic but neutrophilic archaeon, which consisted of only two subunits with relative molecular masses of 66000 and 39000, respectively. PMID- 10681052 TI - Highly efficient purification of porcine diamine oxidase. AB - Diamine oxidase (DAO) is a member of the class of copper-containing amine oxidases and catalyzes the oxidative deamination of histamine and other biogenic amines. The enzyme from porcine kidney was purified by consecutive chromatography on concanavalin A Sepharose, heparin Sepharose and Mono Q. Besides being simpler and faster than previous methods, this new purification scheme results in a homogenous product with a considerably higher yield and allows the rapid purification of large amounts of DAO from mammalian tissues. The availability of sufficient pure protein will greatly facilitate future studies of the structure and function of the enzyme. PMID- 10681053 TI - Development of a downstream process for the isolation of Staphylococcus aureus arsenate reductase overproduced in Escherichia coli. AB - Arsenate reductase (ArsC) encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular As(V) (arsenate) to the more toxic As(III) (arsenite). In order to study the structure of ArsC and to unravel biochemical and physical properties of this redox enzyme, wild type enzyme and a number of cysteine mutants were overproduced soluble in Escherichia coli. In this paper we describe a novel purification method to obtain high production levels of highly pure enzyme. A reversed-phase method was developed to separate and analyze the many different forms of ArsC. The oxidation state and the methionine oxidized forms were determined by mass spectroscopy. PMID- 10681054 TI - Purification of recombinant hydantoinase and L-N-carbamoylase from Arthrobacter aurescens expressed in Escherichia coli: comparison of wild-type and genetically modified proteins. AB - Two enzymes, hydantoinase (HyuH) and L-N-carbamoylase (HyuC), are required for the biocatalytic production of natural and unnatural, optically pure L-amino acids starting from D,L-5-monosubstituted hydantoins using the so called 'hydantoinase-method'. For the preparation of immobilized enzymes, which omit several drawbacks of whole cell biocatalysts, purified or at least enriched HyuH and HyuC have to be provided. In order to simplify existing purification protocols several genetically modified derivatives of HyuH and HyuC from Arthrobacter aurescens DSM 3747 have been cloned and expressed in E. coli. A fusion protein consisting of maltose-binding protein (MalE) and HyuH resulted in an enhanced solubility of the hydantoinase, which easily forms inclusion bodies. On the other hand the fusion protein could easily be purified with high yield (76%) by just one chromatographic step (amylose resin) and the complex purification protocol of the wild-type enzyme could therefore be simplified and shortened significantly. Interestingly, the specific activity of the MalE-HyuH fusion protein was as high as the wild-type enzyme despite that the molecular mass was doubled. A second modification of HyuH carrying a histidine-tag was efficiently bound to a metal affinity matrix but inactivated completely during elution from the column at either low pH or in the presence of imidazole. In the case of HyuC, an aspartate-tag has been added to the biocatalyst to allow an integrated purification-immobilization procedure since this enzyme is immobilized efficiently only via its carboxylic groups. The diminished isoelectric point of the Asp-tagged HyuC resulted in a simplified purification procedure. Compared to the wild-type enzyme expressed in E. coli HyuC-Asp6 was shifted off the elution range of the contaminating proteins and higher purification factors were obtained even in the capturing step. In contrast to HyuH, it was possible to purify a L-N carbamoylase carrying a histidine-tag to apparent homogeneity using immobilized metal affinity chromatography. Therefore, the existing three step purification protocol was reduced to one chromatographic step and the yield of this relatively unstable protein enhanced remarkably. PMID- 10681055 TI - Purification of a D-hydantoinase using a laboratory-scale streamline phenyl column as the initial step. AB - A D-hydantoinase from Thermus sp. was overexpressed in Escherichia coli and purified to homogeneity for subsequent crystallization. The purification was performed with hydrophobic interaction chromatography as the capture step followed by anion-exchange chromatography and gel permeation chromatography as intermediate purification and polishing steps, respectively. The hydrophobic interaction step was done in fluidized bed mode in a laboratory-scale Streamline column made from conventional laboratory equipment. The whole purification protocol could be finished within one day. The purified enzyme crystallizes. The crystals are suitable for X-ray protein structure analysis and diffract to at least 2.3 A resolution. Complete data sets have been measured up to 2.6 A resolution. The X-ray structure is currently being solved. PMID- 10681056 TI - D-Xylose metabolism by Candida intermedia: isolation and characterisation of two forms of aldose reductase with different coenzyme specificities. AB - To study individual enzyme components responsible for the initial step of D xylose utilisation by the yeast Candida intermedia, a two-step protocol has been developed that enables clear-cut separation and isolation of two structurally similar but functionally different aldose reductases (ALRs) in high yield. In the first step, the yeast cell extract is fractionated efficiently by biomimetic chromatography using the dye HE-3B (reactive Red 120) as pseudoaffinity ligand coupled to Sepharose CL-4B. In the second step, optimised high-resolution anion exchange chromatography using Mono Q yields purified ALR1 and ALR2 in overall yields of 63 and 62%, respectively. ALR1 is strictly specific for NADPH (2.4 x 10(5) M(-1) s(-1)) whereas ALR2 utilises NADH and NADPH with similar specificity constants of approximately 2-4 x 10(5) M(-1) s(-1). Both enzymes are dimers with a subunit molecular mass of 36000 but they differ in pI and the number of titratable sulphydryl groups in the native protein. The chromatographic procedure identifies microheterogeneity in recombinant aldose reductase from Candida tenuis overexpressed in Escherichia coli. PMID- 10681057 TI - Purification of surface-associated urease from Helicobacter pylori. AB - Helicobacter pylori colonizes the human gastric mucosa and produces large amounts of urease. The enzyme was extracted from the bacteria by distilled water and purified by gel-permeation (Sephacryl S-300), anion-exchange chromatography (Mono Q) and a second gel-permeation (Superdex 200). Urease enzyme activity was detected with a spectrophotometic assay based on phenol red. The optimal pH for anion-exchange was 6.9. The recovery of urease was 55-75%, purity 93-98% and the overall protein recovery 0.8-1.4%. The urease in the final extract still had enzymatic activity and showed the typical subunits of Mr 66000 and Mr 30000 when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 10681058 TI - Expression of chymotrypsin(ogen) in the thioredoxin reductase deficient mutant strain of Escherichia coli AD494(DE3) and purification via a fusion product with a hexahistidine-tail. AB - A reliable protocol was designed for fast expression and purification of recombinant chymotrypsin(ogen). The zymogen was overexpressed in soluble form as a (His)6-fusion construct in the cytoplasm of the thioredoxin reductase deficient Escherichia coli strain AD494(DE3). This allowed purification of chymotrypsinogen in a highly selective affinity chromatography capture step using a Ni-NTA column. After activation with enterokinase, the enzymatically active chymotrypsin was purified in a polishing step using a modified soybean trypsin inhibitor agarose column. This expression system and the use of affinity chromatography for capture and polishing, offers an easier and faster route to recombinant chymotrypsin(ogen) than the previously described use of Saccharomyces cerevisiae. PMID- 10681059 TI - Refolding and purification of a urokinase plasminogen activator fragment by chromatography. AB - A fragment of recombinant urokinase plasminogen activator (u-PA), was expressed in E. coli in the form of inclusion bodies. Purification and renaturation was achieved in a three-stage process. Capture of the inclusion bodies was achieved by coupling wash steps in Triton X-100 and urea with centrifugation. Solubilised inclusion bodies were then renatured by buffer exchange performed by size exclusion chromatography (SEPROS). Use of size-exclusion media with higher fractionation ranges resulted in an increase in the recovery of u-PA activity, to a maximum fractionation range of Mr 10000-1500000 after which recovery is reduced, due to a low resolution between the refolded u-PA and denaturant. Fractions of refolded u-PA were concentrated using cation ion-exchange chromatography, which selectively binds correctly folded u-PA. The result is concentrated, active, homogeneous u-PA. PMID- 10681060 TI - Purification of human adenosine deaminase for the preparation of a reference material. AB - The goal was to optimise a purification procedure of adenosine deaminase from human erythrocytes for the preparation of a European Reference Material. Adenosine deaminase was purified from human erythrocytes with a specific activity of 4.46 microkat/mg of protein and a catalytic concentration of 133 microkat/l. The isolation and purification procedure involved ion-exchange chromatography (STREAMLINE DEAE), and two purine riboside affinity chromatographies. The purified enzyme exhibits a single band in SDS-PAGE with a molecular weight of 41600 g/mol, and three bands in PAGE, isoelectric focusing and two-dimensional electrophoresis with pI 4.7, 4.85 and 5.0. PMID- 10681061 TI - Pullulanase from the hyperthermophilic bacterium Thermotoga maritima: purification by beta-cyclodextrin affinity chromatography. AB - This is the first report about the isolation of a type I pullulanase from a hyperthermophilic bacterium, Thermotoga maritima strain MSB8. Purification of the enzyme from a cleared cell-free extract was achieved by anion-exchange chromatography and beta-cyclodextrin affinity chromatography. Using this convenient two-step method we have purified the pullulanase 406-fold with a 26% yield. The purified enzyme displayed maximum pullulan hydrolysis at pH 5.9 and 90 degrees C (15-min assay) and was remarkably resistant against thermoinactivation, having a half-life at 90 degrees C of about 3.5 h. To our knowledge, the T. maritima pullulanase is the most thermostable type I pullulanase known to date. The affinity-based purification protocol described here may be useful for the efficient isolation of other pullulanases. PMID- 10681062 TI - Single-step purification of a recombinant thermostable alpha-amylase after solubilization of the enzyme from insoluble aggregates. AB - The expression of the gene encoding a thermostable alpha-amylase (EC 3.2.1.1) (optimal activity at 100 degrees C) from the hyperthermophilic archaeon Pyrococcus woesei in the mesophilic hosts Escherichia coli and Halomonas elongata resulted in the formation of insoluble aggregates. More than 85% of the recombinant enzyme was present within the cells as insoluble but catalytically active aggregates. The recombinant alpha-amylase was purified to homogeneity in a single step by hydrophobic interaction chromatography on a phenyl superose column after solubilization of the enzyme under nondenaturing conditions. The enzyme was purified 258-fold with a final yield of 54%. PMID- 10681063 TI - Technique for a rapid and efficient purification of the SHV-1 and PSE-2 beta lactamases. AB - A simple procedure is described which results in an optimised resolution in molecular sieve chromatography. A sample exhibiting a large initial volume (about 20 ml) and conditioned in a buffer of low ionic strength (<20 mM) by filtration through a 53-ml G25 molecular sieve column, is adsorbed on a 1.7-ml ion-exchange (SOURCE) column. The proteins are released by a 10-ml pulse of 1 M NaCl and the eluate directly injected onto a 120-ml Sephacryl S100-HR column. The very low volume of the eluate ensures optimal conditions and resolution for the molecular sieving process. The method is applied as the polishing step in the purification of the SHV-1 and PSE-2 beta-lactamases. It could easily be scaled up for the treatment of larger samples. PMID- 10681064 TI - Purification of the fengycin synthetase multienzyme system from Bacillus subtilis b213. AB - The purification of the multienzyme system producing the lipodecapeptide fengycin in Bacillus subtilis b213 was investigated. By gel filtration of a cell free extract of this organism three enzyme fractions were obtained from which five multifunctional components of fengycin synthetase were separated by high resolution anion-exchange FPLC procedures. These proteins were characterized by their thioester formation activities with 14C-labeled substrate amino acids and by N-terminal sequencing. Correlation of these data with the DNA sequences of the pps (fen) operons in three B. subtilis strains provided detailed knowledge on the structural and functional organization of fengycin synthetase. PMID- 10681065 TI - New approach for separating Bacillus subtilis metalloprotease and alpha-amylase by affinity chromatography and for purifying neutral protease by hydrophobic chromatography. AB - Proteases are commonly used in the biscuit and cracker industry as processing aids. They cause moderate hydrolysis of gluten proteins and improve dough rheology to better control product texture and crunchiness. Commercial bacterial proteases are derived from Bacillus fermentation broth. As filtration and ultrafiltration are carried out as the only recovery steps, these preparations contain also alpha-amylase and beta-glucanase as the main side activities. The aim of this study is to purify and characterize the Bacillus subtilis metalloprotease from a commercial preparation, in order to study separately the impact of the protease activity with regards to its functionality on biscuit properties. Purification was achieved by means of affinity chromatography on Cibacron Blue and HIC as a polishing step. Affinity appeared to be the most appropriate matrix for large scale purification while ion exchange chromatography was inefficient in terms of recovery yields. The crude product was first loaded on a Hi Trap Blue column (34 microm, Pharmacia Biotech); elution was carried out with a gradient of NaCl in the presence of 1 mM ZnCl2. This step was only efficient in the presence of Zn cations, because this salt promoted both protease stabilization resulting in high recovery yields and also complexation of amylase units into dimers resulting in amylase retention on the column and a better separation of the 3 activities. Beta-glucanase was mostly non retained on the column and a part was coeluted with the protease. This protease fraction was then loaded on a Resource Phe column (15 microm, Pharmacia Biotech) in a last step of polishing. Elution was carried out with a linear gradient of 100-0% ammonium sulfate 1.3 M; protease was eluted at the beginning of the gradient and well separated from amylase and glucanase trace impurities. The homogeneity of the purified protease was confirmed by SDS-PAGE, which showed that its MW was about 38. pH and temperature optima were also determined on the fraction. PMID- 10681066 TI - Purification and characterization of an aminopeptidase from the chloroplast stroma of barley leaves by chromatographic and electrophoretic methods. AB - Aminopeptidases catalyze the cleavage of amino acids from the amino terminus of protein or peptide substrates. Although some aminopeptidase activities have been found in plant chloroplasts, the identity of these proteins remains unclear. In this work, we report the purification to apparent homogeneity of a soluble aminopeptidase from isolated barley chloroplasts which preferentially degraded alanyl-p-nitroanilide (Ala-pNA). After organelle isolation in a density gradient and precipitation of soluble proteins with ammonium sulfate, the proteins were purified in three consecutive steps including hydrophobic interaction, gel permeation and ion-exchange chromatographies. The purified enzyme appeared as a single band with a Mr of approximately 84000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The Mr of the native enzyme was estimated to be approximately 93000 by gel permeation chromatography, suggesting that the protein is a monomer. Mass spectrometry analysis of tryptic digests indicates that the primary structure of the protein has not been reported previously. The enzyme was characterized as a metalloprotease as it could be totally inhibited by 1,10-phenanthroline. Strong inhibition could also be observed using the specific aminopeptidase inhibitors amastatin and bestatin. Besides Ala-pNA, the purified protein could also cleave with decreasing activity glycyl-pNA, leucyl-pNA, lysyl-pNA, methionyl-pNA and arginyl-pNA. The possible physiological role of this enzyme in the chloroplast stroma is discussed. PMID- 10681067 TI - Three-step chromatographic purification of Cpr6, a cyclophilin from Saccharomyces cerevisiae. AB - Cyclophilins constitute a group of peptidyl-prolyl cis-trans isomerases (PPIs), known to be involved in protein folding. Because of their ability to bind the immunosuppresant drug Cyclosporin A (CsA), they are also called immunophilins. Immunophilins, which exhibit a relative molecular mass higher than 40 000, are further found in complex with Hsp90, a major cytosolic molecular chaperone. The present work describes a three-step chromatographic purification of recombinant Cpr6, a cyclophilin from Saccharomyces cerevisiae. The cDNA of Cpr6 was cloned into a pRSET A-plasmid with an N-terminal 6 x histidine-tag (his-tag) and transformed into the BL21[DE3]pLysS strain. After collection of the bacterial material and lysis of the cells the cell lysate was centrifuged and loaded onto a metal chelating column. After extensive washing the protein was eluted with a step gradient from 20 to 250 mM imidazol. The pooled protein was dialysed against ethylenedinitrilo tetraacetic acid (EDTA)-buffer, and loaded onto a strong anion exchanger. Cpr6 containing fractions were then, in a last step, loaded onto a gel permeation chromatography column. The purity of the resulting protein was measured by silver stained sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and, additionally, as Cpr6 does not contain tryptophan residues by tryptophan residue titration. Based on a standard curve the content of contaminating tryptophan residues in the purified protein solution was determined. A typical yield of 1 mg pure protein per g of wet cells was achieved with the described procedure. PMID- 10681068 TI - Are isoprostanes a clinical marker for antioxidant drug investigation? AB - Numerous pathological conditions are suspected to involve free radical production as part of their pathogenic process. Therefore, a pharmacological control of oxidative stress could probably benefit many vascular, inflammatory or degenerative diseases. However, the development of antioxidant drugs and their clinical evaluation are limited by the absence of an accurate, reliable and easy to-handle marker of tissue oxidative events. Isoprostanes (isoPs), a prostaglandin-related series of metabolites, are emerging as major candidates for clinical measurement of oxidative stress. They are chemically stable products of lipid peroxidation, formed in cellular membranes and subsequently released and excreted in the urine. Many recent clinical studies have reported that urinary and plasma levels of isoPs (in particular the iPF2alpha-III isomer also called 8 epi-PGF2alpha) are increased in clinical conditions where oxidative stress is suspected to play a pathogenic role. Moreover, isoPs have been detected in tissue extracts from atherosclerotic plaques and Alzheimer patients brain tissue. Finally, antioxidant treatments such as vitamin E supplementation appear to reduce isoPs levels in biological fluids of treated patients. These preliminary observations argue for a further investigation of isoPs as a practical pharmacodynamic endpoint for the clinical evaluation of antioxidant therapies. PMID- 10681069 TI - Effects of non-ionic monomeric and dimeric iodinated contrast media on renal and systemic haemodynamics in rats. AB - Non-ionic dimeric contrast media (CM) are a new class of CM which are iso-osmolar with plasma. The aim of this study was to investigate their effects on systemic and renal haemodynamics. The non-ionic dimeric CM iodixanol and the non-ionic monomeric agent iobitridol (both at a dose of 1,600 mgI/kg) were compared in terms of their effects on systemic blood pressure (BP) and renal blood flow (RBF) in two strains of rats (Wistar and Sprague Dawley). Iodixanol significantly lowered BP in Wistar rats (-33 +/- 9% of baseline, 10 min post-injection, P < 0.001 vs. saline and iobitridol). Iobitridol had virtually no effect on BP. Iobitridol and iodixanol significantly decreased RBF. This effect was more marked following injection of the dimer rather than the monomer (iodixanol: -32 +/- 13% iobitridol: -20 +/- 4 of baseline at 16 min, P < 0.05). For both agents, RBF was still decreased 50 min following injection (iodixanol: -30 +/- 11%, and iobitridol: -20 +/- 5% of baseline). Iodixanol also decreased RBF in Sprague Dawley rats, while BP remained unchanged. This suggests that changes in BP/RBF autoregulation do not account for the renal haemodynamic effects of this agent. When measured 2 h following injection, the iodixanol-induced renal hypoperfusion was still detectable (-29% vs. saline-treated rats), although not significant (P = 0.06). This effect was no longer observed 4 h following injection. Increasing the saline infusion rate (18 mL/h vs. 2 mL/h) during the experiment did not significantly decrease the effects of iodixanol on BP and RBF in Wistar rats. In spite of its iso-osmolality, iodixanol, a non-ionic dimeric CM, depressed RBF and BP significantly more than iobitridol, a monomeric non-ionic agent, in Wistar rats. This effect was long-lasting and was not alleviated by increasing the hydration rate. PMID- 10681070 TI - Detection of salicylate and its hydroxylated adduct 2,3-dihydroxybenzoic acid in glutamate neurotoxicity and the effects of verapamil and ryanodine in rats. AB - It has been suggested that salicylate hydroxylation can be used to detect hydroxyl radical formation in vivo. In the present study we investigated the effects of verapamil and or ryanodine on salicylate (SA) and its hydroxylated adduct; 2,3-dihydroxybenzoic acid (2,3-DHBA) levels in glutamate induced neurotoxicity of whole rat brains. To detect SA and 2,3-DHBA, an HPLC-EC/UV method was used. Retention time was found to be 3.9 min for 2,3-DHBA and 12.0 min for SA. Verapamil at 10(-5) and 10(-7) and ryanodine at 10(-5) M concentrations were found to have a significant decreasing effect on this degradation induced by glutamate. This was the highest dose for ryanodine tested. As an L-type voltage dependent calcium channel blocker, verapamil was found ineffective at 10(-4), 10( 6) and 10(-8) M concentrations. Surprisingly, none of the combined application groups (verapamil + ryanodine) was found effective on SA hydroxylation. As a result, ryanodine was effective only at the highest dose, while verapamil exerts its effect in a dose dependent fashion as reported before in the literature. PMID- 10681071 TI - Role of the endothelium on the response to adrenoceptor agonists of rabbit aorta during cooling. AB - The role of the endothelium in the effects of cooling on the response to alpha1- and alpha2-adrenoceptor agonists of rabbit aorta was studied. The contractions induced by clonidine (10(-9)-3 x 10(-4) M) and xylazine (10(-7)-10(-3) M) but not phenylephrine (10(-9)-3 x 10(-4) M) and methoxamine (10(-9)-3 x 10(-4) M) were enhanced in endothelium-denuded or N(G)-nitro-L arginine methyl- ester (L-NAME) (10(-5) M) pretreated rabbit aorta. The sensitivity, but not the maximal response, of both alpha1- and alpha2-adrenoceptor agonists was significantly lower at 28 degrees C (cooling) than at 37 degrees C. Endothelium removal did not affect the action of cooling. These results were taken as evidence for the specificity of alpha2-adrenoceptor agonists on the production and release of nitric oxide from vascular endothelium. The results suggest that the endothelium seems to have no role in the cooling-induced responses of both alpha1- and alpha2 adrenoceptors. PMID- 10681072 TI - Comparison of the efficacy and safety of losartan (50-100 mg) with the T-type calcium channel blocker mibefradil (50-100 mg) in mild to moderate hypertension. AB - The objective of this study was to compare the antihypertensive efficacy and safety of losartan and mibefradil. 324 outpatients (57 +/- 9.2 years) with mild to moderate hypertension were randomly allocated in a double-blind fashion to receive 50 mg of losartan or mibefradil once daily p.o. for 6 weeks after 2 weeks of placebo run-in. Titration was then forced to 100 mg of losartan or mibefradil for an additional 6 weeks. Patients were assessed at baseline, 6 and 12 weeks. The primary efficacy variable was change in predose sitting diastolic (SDBP) and systolic (SSBP) blood pressure at 12 weeks. Secondary variables included change in mean 24-hour ambulatory blood pressure and comparison of safety and tolerability. Both treatments lowered SSBP and SDBP at 6 and 12 weeks (week 6: mibefradil -14/-9 mm Hg; losartan -12/-7 mm Hg) (P <0.001). The primary objective, a difference between treatments in reduction of SSBP and SDBP at week 12 could be demonstrated (mibefradil -22/-16 mm Hg; losartan -16/-10 mm Hg) (P=0.003 and P=0.001, respectively). Twenty-four-hour SBP and 24-hour DBP were reduced (P<0.001) within each treatment group at weeks 6 and 12. The secondary objective, a difference between treatments in reduction of 24-hour blood pressure at week 12 could be demonstrated (P<0.001). Twenty-four-hour heart rate was lowered in the mibefradil group at weeks 6 and 12 (P < 0.001). Responder rates at 6 and 12 weeks were 56.2% and 78.5% for mibefradil versus 56.1% and 55.3% for losartan (P = 0.001). Both treatments were equally well tolerated. This study demonstrates that 50 mg losartan is comparably effective to 50 mg mibefradil in the treatment of mild to moderate hypertension with 100 mg mibefradil being more potent than losartan. PMID- 10681073 TI - Criteria for vascular dementia: replacing dogma with data. PMID- 10681074 TI - Alzheimer disease: mouse models pave the way for therapeutic opportunities. AB - Research into the molecular mechanisms of Alzheimer disease (AD) continues to clarify important issues in aberrant protein processing while seeking to identify therapeutic targets. Mutations of genes on chromosomes 1, 14 (presenilins 1 and 2), and 21 (the amyloid-beta [Abeta] amyloid precursor protein [APP]) cause the familial forms of AD that often begin before age 65. An allelic polymorphism on chromosome 19 (apolipoprotein E ) affects the age of onset of the more common forms of sporadic AD. Multiple studies in transgenic mice provide strong evidence to support the view that Abeta amyloid formation is an early and critical pathogenic event: mice expressing pathogenic human APP mutations develop Abeta deposits; coexpression of mutant presenilin genes accelerates the rate of Abeta deposition; and apolipoprotein E plays a role in this process. Thus, the 3 established genetic causes or risk factors for AD affect Abeta deposition. The fact that elevation of the Abeta42/Abeta40 ratio (differing only in 2 amino acids in length) is also linked to amyloid deposition in the APP mice and is temporally linked to cognitive impairment suggests that Abeta42 may be a principal inducing factor of AD. The exact sequence of events is still unknown, but the transgenic models generated so far have shown their usefulness in clarifying this complex part of the pathology. The continuing progress in elucidation of the molecular pathogenesis of AD suggests a range of rational pharmacological interventions for this disorder. The most promising strategy involves the development of approaches to retard, halt, or prevent Abeta-mediated disease progression, and these can now be tested in transgenic animals. PMID- 10681075 TI - Cortical and subcortical interhemispheric interactions following partial and complete callosotomy. PMID- 10681076 TI - Clinical criteria for the diagnosis of vascular dementia: a multicenter study of comparability and interrater reliability. AB - BACKGROUND: Several clinical criteria have been developed to standardize the diagnosis of vascular dementia (VaD). Significant differences in patient classification have been reported, depending on the criteria used. Few studies have examined interrater reliability. OBJECTIVE: To assess the concordance in classification and interrater reliability for the following 4 clinical definitions of VaD: the Hachinski Ischemic Score (HIS), the Alzheimer Disease Diagnostic and Treatment Centers (ADDTC), National Institute of Neurological Disorders and Stroke-Association Internationale pour la Recherche et l'Enseignement en Neurosciences (NINDS-AIREN), and Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). METHODS: Structured diagnostic checklists were developed for 4 criteria for VaD, 2 criteria for Alzheimer disease (AD), and 4 criteria for dementia. Twenty-five case vignettes, representing a spectrum of cognitive impairment and subtypes of dementia, were prepared in a standardized clinical format. Concordance in case classification using different criteria and interrater reliability among 7 ADDTCs given a specific set of criteria was assessed using the kappa statistic. RESULTS: The frequency of a diagnosis of VaD was highest using the modified HIS or DSM-IV criteria, intermediate using the original HIS and ADDTC criteria, and lowest using the NINDS-AIREN criteria. Scores for interrater reliability ranged from kappa = 0.30 (ADDTC) to kappa = 0.61 (original HIS). CONCLUSIONS: Clinical criteria for VaD are not interchangeable. Depending on the criteria selected, the reported prevalence of VaD will vary significantly. The traditional HIS has higher interrater reliability than the newer criteria for VaD. Prospective longitudinal studies with clinical-pathological correlation are needed to compare validity. PMID- 10681077 TI - Effects of bilateral posteroventral pallidotomy on gait of subjects with Parkinson disease. AB - BACKGROUND: Most studies documenting the effect of pallidotomy on parkinsonian gait have reported unilateral surgery and used qualitative scales or timed tests that only provide measures of walking speed. OBJECTIVE: To document the effect of bilateral posteroventral pallidotomy on the walking patterns of patients with Parkinson disease (PD). DESIGN: Case series of gait evaluations performed 1 month before and 1 month after surgery, with antiparkinson medication withheld for 8 hours overnight. SETTING: Movement analysis laboratory of a clinical research center. PATIENTS: Consecutive sample of 8 men and 3 women with a diagnosis of PD scheduled for bilateral pallidotomy. INTERVENTION: Bilateral posteroventral pallidotomy. MAIN OUTCOME MEASURES: A 3-dimensional motion-capture system allowed calculation of temporal and spatial measurements and joint angular displacements of the lower extremities and trunk during gait. RESULTS: Pallidotomy significantly increased average walking speed from 0.214 statures/s preoperatively to 0.440 statures/s postoperatively (where stature indicates body height) (P = .03). A faster postoperative walking speed was achieved almost exclusively by increasing average stride length from 0.24 to 0.47 statures (P = .03) rather than changing average gait cycle time (1.32 to 1.37 seconds; P = .08). A forward stepwise multiple regression analysis (P<.001) revealed that 96% of the change in stride length postoperatively could be explained by the combination of changes in foot-floor angle, knee, and hip excursion during gait. CONCLUSIONS: Bilateral posteroventral pallidotomy was associated with a 2-fold increase in walking speed. Previous studies have demonstrated that walking speed is an important indicator of locomotor performance and level of disability in patients with PD, so the increase in postoperative walking speed likely provided a functional benefit. PMID- 10681078 TI - Striatal dopamine transporter binding assessed by [I-123]IPT and single photon emission computed tomography in patients with early Parkinson's disease: implications for a preclinical diagnosis. AB - BACKGROUND: Specific binding to dopamine transporters may serve as a tool to detect early loss of nigrostriatal dopaminergic neurons in patients with Parkinson's disease. OBJECTIVE: To determine striatal dopamine transporter binding using the cocaine analogue [I-123]N-(3-iodopropen-2-yl)-2beta carbomethoxy-3beta-(4-chl orophenyl) tropane ([I-123]IPT) and single photon emission computed tomography. PATIENTS AND METHODS: We studied 9 control subjects (mean age, 58 years; range, 41-69 years) and 28 patients with early Parkinson's disease (Hoehn and Yahr stages I [n = 14] and II [n = 14] [symptom duration, <5 years]; mean age, 55.5 years; range, 36-71 years). Single photon emission computed tomography was performed 90 minutes after injection of 120 to 150 MBq of radioactive [I-123]IPT. RESULTS: Specific striatal [I-123] IPT binding (mean +/- SD) was significantly reduced in patients with early Parkinson's disease (ipsilateral striatum: 4.09+/-0.97; range, 2.46-6.40; contralateral striatum: 3.32+/-0.76; range, 1.80-5.13) compared with controls (left striatum: 7.28+/ 0.94; range, 5.78-8.81; right striatum: 7.41+/-1.28; range, 5.58-9.44). IPT binding ratios (mean +/- SD) were significantly lower in patients with Hoehn and Yahr stage II (ipsilateral striatum: 3.47+/-0.75; contralateral striatum: 2.96+/ 0.73) compared with those with Hoehn and Yahr stage I (ipsilateral striatum: 4.72+/-0.75; contralateral striatum: 3.69+/-0.61) (P<.001). The ipsilateral striatum of patients with Hoehn and Yahr stage I showed a significant mean+/-SD reduction of IPT binding (ipsilateral striatum: 4.72+/-0.75) compared with either right or left striatum of controls (P<.001). Only in 1 patient was IPT binding to the ipsilateral striatum (ratio, 6.40) higher than the lowest value observed in the striatum of a control subject (ratio, 5.58). CONCLUSIONS: Use of [I-123] IPT and single photon emission computed tomography demonstrates a reduction of dopamine transporter binding in patients with early Parkinson's disease. Significantly reduced IPT binding already observed in the ipsilateral striatum of patients with Hoehn and Yahr stage I demonstrates the potential of this method to detect preclinical disease. PMID- 10681079 TI - Association between angiotensin-converting enzyme and Alzheimer disease. AB - BACKGROUND: Angiotensin-converting enzyme has been reported to show altered activity in patients with neurologic diseases. An insertion-deletion polymorphism in ACE has recently been linked to heart disease, cerebrovascular disease, and AD. OBJECTIVE: To determine whether the angiotensin-converting enzyme (ACE) is associated with risk of Alzheimer disease (AD). METHODS: We investigated the ACE polymorphism as a potential risk factor for AD in 151 patients with AD and 206 ethnically matched controls from Russia and in 236 patients with AD and 169 controls from North America by means of allele association methods and logistic regression. RESULTS: None of the ACE genotypes was associated with increased susceptibility to AD in the total sample or in subsets stratified by apolipoprotein E gene (APOE) epsilon4 status. However, the D allele was more frequent among AD cases between ages 66 and 70 years compared with controls in both the Russian (P = .02) and North American (P = .001) datasets. In this age group, the effect of D (odds ratio, 11.2; 95% confidence interval, 2.9-44.0) appeared to be independent of and equal or greater in magnitude to the effect of APOE epsilon4 (odds ratio, 7.8; 95% confidence interval, 3.5-7.4). CONCLUSIONS: Our results suggest that APOE and ACE genotypes may be independent risk factors for late-onset AD, but the ACE association needs to be confirmed in independent samples in which the time and extent of vascular cofactors can be assessed. PMID- 10681080 TI - A novel missense mutation (W797R) in the myophosphorylase gene in Spanish patients with McArdle disease. AB - OBJECTIVE: To investigate the degree of genetic heterogeneity of myophosphorylase deficiency (McArdle disease) in Spain through molecular studies of 10 new patients. DESIGN: The coding sequence of the entire myophosphorylase gene was sequenced in DNA extracted from muscle and blood. Restriction fragment length polymorphism analysis of polymerase chain reaction fragments was used to confirm and simplify detection of a novel mutation. SETTING: A collaborative study between 2 university laboratories in Spain and the United States. RESULTS: Five of the 10 patients harbored a novel missense mutation in exon 20, converting a tryptophan to an arginine (W797R). Three patients were homozygous for the "common" R49X mutation, and the remaining 2 patients were compound heterozygotes for R49X and a previously described missense mutation, G204S. CONCLUSIONS: The W797R missense mutation is the third novel mutation to be identified among Spanish patients. Its relative frequency suggests that it should be added to the R49X mutation in the molecular screening of McArdle disease in Spain. PMID- 10681081 TI - Intracranial volume and Alzheimer disease: evidence against the cerebral reserve hypothesis. AB - BACKGROUND: Total intracranial volume (TIV) measurement commonly is used to correct for variations in premorbid brain size in imaging studies of cerebral structures in Alzheimer disease (AD). This assumes no intrinsic difference in TIV between patients and control subjects and that TIV measurements are unaffected by cerebral atrophy. However, an autopsy study has suggested that a larger premorbid brain may protect against AD onset. A recent computed tomographic study lent support to this by finding a correlation between intracranial size and age at onset of AD in women. OBJECTIVE: To investigate the relationship between TIV and sporadic and familial AD. DESIGN: Retrospective case study. SETTING: Specialist dementia clinic. PATIENTS: Eighty-five patients with AD and 52 healthy volunteers. MAIN OUTCOME MEASURES: Age at symptom onset and TIV measured using a semiautomatic interactive thresholding technique on magnetic resonance imaging spanning the entire intracranial cavity. RESULTS: Reproducibility measurement was high (intrarater coefficient of variation, 1.2%; interrater coefficient of variation, 0.7%). Unlike brain atrophy in the patients with AD, TIV did not vary over time. Mean TIV did not differ significantly between any of the subject groups. There was no association between TIV and age or age at symptom onset. The only significant predictor of TIV was sex. CONCLUSIONS: Measurements of TIV are independent of atrophy and can be used safely to adjust for differences in head size in studies of cerebral structure in AD. Premorbid brain size does not differ between patients with familial and sporadic AD and controls and does not delay disease onset. PMID- 10681082 TI - Differences between Pick disease and Alzheimer disease in clinical appearance and rate of cognitive decline. AB - OBJECTIVES: To define the cognitive characteristics of Pick disease (PcD), and to determine which features distinguish PcD from Alzheimer disease (AD), in a cross sectional and longitudinal study. METHODS: The participants were 44 patients with PcD (10 pathologically verified), 121 patients with AD (14 pathologically verified), and 60 normal control subjects. We obtained information regarding the initial symptom of dementia from each patient's caregiver, estimated global dementia severity by the Blessed Dementia Scale and the Activities of Daily Living Scale, and assessed specific cognitive domains by administering 10 tests of memory, language, visuospatial, and reasoning abilities and selective attention. RESULTS: Among initial symptoms reported by caregivers, personality change and language impairment were significantly more common in PcD than AD; deficits in memory were common in both groups but more prevalent in AD (P<.001). At initial cognitive testing, the scores of patients with PcD were inferior to those of normal controls on all tests, except on a measure of visuospatial function; the scores of patients with AD were inferior to those of controls on all tests. Patients with PcD were superior to patients with AD on measures of explicit memory (P<.001) and visuospatial function (P = .001) but had greater impairments on the Activities of Daily Living Scale (P<.05). During the course of illness, patients with PcD declined significantly faster than those with AD on language tests and on global measures of dementia severity (P<.05), whereas measures of explicit memory and visuospatial and reasoning abilities worsened equally in both patient groups. CONCLUSIONS: There is a characteristic cognitive profile and course of dementia in PcD. Nonetheless, cognitive test performance does not clearly distinguish PcD from AD. PMID- 10681083 TI - Lack of an association of estrogen receptor alpha gene polymorphisms and transcriptional activity with Alzheimer disease. AB - BACKGROUND: Long-term cognitive decline in postmenopausal women is associated with aging and Alzheimer disease (AD). Estrogen replacement therapy has been reported to reduce the risk of developing AD. The distribution of estrogen receptors (ERs) in neurons overlaps that of the brain neurons known to develop AD. Estrogen increases the secretion and metabolism of amyloid precursor protein, may help synapse formation, and is reported to protect neurons from toxins. Restriction fragment length polymorphisms (RFLPs) of the ERalpha gene at intron 1 and exon 2 were associated with a low bone mineral density in postmenopausal women and also with AD in a Japanese population. OBJECTIVE: To determine whether ERalpha gene polymorphisms are associated with transcriptional activity and AD. METHODS: A luciferase reporter assay analyzed enhancer activity of the ERalpha gene at intron 1 and exon 2. This activity was evaluated according to the RFLPs. The RFLPs of the ERalpha gene were determined in Japanese patients clinically diagnosed as having AD, white patients diagnosed as having AD at autopsy, and corresponding healthy control subjects. The RFLPs were also evaluated for the contribution of the ERalpha gene RFLPs to AD. RESULTS: We found weak (about 2 fold) enhancer activity of the ERalpha gene, which differed among RFLPs. Although there were racial differences in these polymorphisms, we could not confirm the previously reported association between ERalpha gene polymorphisms and AD. CONCLUSION: Regulatory element of the ERalpha gene was found in intron 1, but we found no association between ERalpha gene polymorphisms and AD. PMID- 10681084 TI - Very late-onset Friedreich ataxia despite large GAA triplet repeat expansions. AB - BACKGROUND: Most patients with Friedreich ataxia (FRDA) have abnormal GAA triplet repeat expansions in both X25 genes. The size of the GAA expansion in the shorter of the 2 expanded alleles correlates significantly with parameters of clinical severity and is inversely related to the age at onset. OBJECTIVES: To describe the clinical and molecular genetic findings in a patient with very late-onset FRDA and to review the literature. PATIENT AND METHODS: A 58-year-old white woman with mild progressive gait disturbance of 15 years' duration whose examination revealed mild incoordination was analyzed for mutations in the X25 gene. A combination of long-range polymerase chain reaction and genomic Southern blot analyses were used to identify GAA expansions in intron 1 of the X25 gene. To uncover evidence of somatic variability in triplet repeat length, DNA isolated from several tissue samples was similarly analyzed. Single-strand conformational polymorphism analysis was used to screen for mutations spanning the entire coding sequence of frataxin and all intron-exon junctions of the X25 gene. RESULTS: DNA isolated from blood leukocytes revealed GAA triplet repeat expansions in both X25 genes, which were estimated to contain 835 and 1200 repeats. Similar expansions were detected in DNA isolated from lymphoblasts, fibroblasts, buccal cells, and sural nerve, with estimated mean (+/- SD) lengths of the shorter and longer expansions being 854 (+/-69) and 1283 (+/-72) triplets, respectively. A review of reported cases of late-onset Friedreich ataxia (25-39 years) and very late-onset Friedreich ataxia (> or =40 years) demonstrated that this is the first instance of a patient presenting with very late-onset FRDA despite carrying more than 800 GAA repeats in both expanded X25 alleles. CONCLUSIONS: This unique case of very late-onset FRDA highlights a limitation in our ability to accurately predict the phenotype in FRDA based solely on the size of the GAA expansion. Other genetic or environmental factors may significantly modify disease severity in FRDA. PMID- 10681085 TI - Mechanism in progressive lacunar infarction: a case report with magnetic resonance imaging. AB - BACKGROUND: The mechanism of a progressive lacunar infarction is not well understood, and changes in ischemic tissue after onset have not yet been clarified clinically. OBJECTIVE: To investigate the pathophysiological characteristics of a case of progressive lacunar infarction using diffusion weighted and conventional magnetic resonance imaging (MRI) scans. PATIENT: A 73 year-old woman was hospitalized 18 hours after stroke onset and was diagnosed as having a lacunar infarction in the perforating territory of the left middle cerebral artery. Despite treatment, the hemiparesis worsened, with the peak on the fourth day after onset. Diffusion-weighted and conventional MRI scans provided clues to the pathogenesis. FINDINGS AND CONCLUSIONS: In the acute stage, gradual enlargement of the hyperintense lesion, reflecting fresh ischemic tissue, and neurological deterioration were observed by serial examination of diffusion weighted MRI scans. A conventional coronal MRI scan revealed a 2-layered ischemic lesion, suggesting the involvement of perforating arteries. These findings indicated that hemodynamic impairment of the microcirculation in the perforators was the major cause of the lacunar infarction. PMID- 10681086 TI - Cerebral venous thrombosis associated with epoetin alfa therapy. AB - BACKGROUND: Cerebral venous thrombosis is a rare complication of polycythemia. To our knowledge, epoetin alfa-induced polycythemia has not previously been reported in association with cerebral venous thrombosis. CASE DESCRIPTION: A 37-year-old patient who was receiving peritoneal dialysis and epoetin alfa (Epogen) therapy presented with a several-day history of worsening headache, and a neuroimaging scan demonstrated thrombosis of the sagittal and transverse sinus. Epoetin alfa therapy, which had been initiated 3 months earlier according to an institutional protocol, was associated with a problematic increase in hematocrit values. CONCLUSIONS: Headache should raise the suspicion of cerebral venous thrombosis in patients who are being treated with epoetin alfa, particularly in the presence of elevated hematocrit values. Monitoring hematocrit parameters in accordance with "standard guidelines" is recommended. PMID- 10681087 TI - Tolcapone and hepatotoxic effects. Tasmar Advisory Panel. AB - Four patients with Parkinson disease have recently been described in whom severe hepatic dysfunction developed in association with tolcapone therapy. These reports led to the introduction of a "black box" warning and more intensive monitoring requirements in the United States. A review of these cases and all clinical trials indicates that liver dysfunction did not develop in any patient who had received monitoring of liver function according to the original prescribing information. Virtually all instances of liver enzyme abnormality and clinical liver dysfunction occurred within 6 months of initiating treatment. To assess the current role of tolcapone therapy in Parkinson disease, a panel of neurologists and hepatologists was convened. Consensus was reached with respect to the following: (1) Tolcapone is an effective agent in the treatment of patients with fluctuating Parkinson disease. (2) The risk of developing irreversible liver injury is negligible with appropriate monitoring. (3) It may be possible to reduce the frequency of monitoring after 6 months of treatment. (4) The requirement that tolcapone be withdrawn if liver enzymes are elevated above the upper limit of normal on a single occasion is unnecessarily restrictive. It was concluded that tolcapone, when used as an adjunct to levodopa, is an effective anti-parkinsonian agent and that less frequent monitoring after 6 months, with an action limit of 2 to 3 times the upper limit of normal, is sufficient to ensure safety in patients who are deriving benefit from the drug. PMID- 10681088 TI - Weak and numb feet in a man with knobby hands. PMID- 10681089 TI - New drugs: which should be included in the formulary? Epilepsy: all new drugs should be included. PMID- 10681090 TI - Restrictions on the availability of antiepileptic drugs. PMID- 10681091 TI - New antiepileptic drugs: should all be made available? PMID- 10681092 TI - Wilson's disease. PMID- 10681093 TI - Solvent toxicity and cognition impairment. PMID- 10681094 TI - Effects of hormone replacement therapy on lipid peroxides and oxidation system in postmenopausal women. AB - A short-term evaluation of 6 months of estrogen therapy on oxidant status in 38 postmenopausal women was conducted. The levels of serum lipid peroxidation products, glutathione (GSH) status, and glutathione-related enzymes were evaluated before and after 6 months of hormone replacement therapy. After 6 months of estrogen treatment there was a significantly increased concentration of thiobarbituric acid-reactive substances (TBARS), which are an end product of lipid peroxidation. This was accompanied by a significant increase in the activity of glutathione peroxidase (GSH-Px). However, the activities of glutathione reductase (GSSG-R) and superoxide dismutase (SOD) were significantly decreased and total protein thiols were reduced. Data suggest that hormone replacement therapy in postmenopausal women is associated with oxidant mechanisms. PMID- 10681095 TI - Comparison of oxidative stress indicators in plasma of recent-onset and long-term type 1 diabetic patients. AB - Oxidative stress was compared in plasma of 15 recently diagnosed (<2 mo) or 15 longstanding (>5 yr) type 1 diabetic patients with 15 healthy volunteers. Lipid peroxidation indices measured in plasma included thiobarbituric acid-reactive substances (TBARS), conjugated dienes, and lipid hydroperoxide (ROOH). The values obtained were corrected for phospholipid to minimize this as a confounding factor. In recently diagnosed diabetics, plasma conjugated lipid dienes were significantly elevated. However, in longstanding diabetics there was a marked increase in TBARS, conjugated dienes, and lipid hydroperoxide levels. Our findings showed increased oxidative stress in type 1 diabetics regardless of metabolic control and that conjugated diene measurement appeared to be the most sensitive bioindicator of oxidant stress in our population. PMID- 10681096 TI - Are individuals with glutathione S-transferase GSTT1 null genotype more susceptible to in vitro oxidative damage? AB - Recent epidemiological studies proposed that glutathione S-transferase (GST) T1 null genotype was correlated with an increased susceptibility to diseases associated with oxidative stress, including cancer. A comparative study using erythrocytes from individuals with GSTT1 null genotype was carried out to determine how resistance to oxidative stress is affected by lack of this gene, and whether the GST status of a person is an important factor in risk toward oxidant chemicals. Malondialdehyde and carbonyl levels and fluorescence and chemiluminescence formation were used as biomarkers of oxidative stress in erythrocytes exposed in vitro to cumene hydroperoxide (CumOOH), an oxidizing agent. When peroxidation-dependent changes in these parameters were compared between GSTT1 null genotype and controls, who are both GSTM1 and GSTT1 positive, no significant differences were found between the two genotypes, although the erythrocytes of the GSTT1 null group had lower GSTT1 activity toward CumOOH. Our results indicate that erythrocytes from individuals with GSTT1 null genotype are not abnormally susceptible to CumOOH-induced oxidant challenge. PMID- 10681097 TI - Embryotoxic and teratogenic effects of indium chloride in rats and rabbits. AB - Daily indium chloride doses of control (0), 50, 100, 200, or 400 mg/kg were administered orally to Sprague-Dawley rats by gavage, on d 6-15 of gestation, and daily metal doses of control (0), 50, 100, or 200 mg/kg were administered to New Zealand rabbits on d 6-20 of gestation. Further groups of pregnant rats were treated with control (0) or 400 mg/kg indium chloride orally on one of d 8, 9, 10, 11, 12, 13, 14, or 15 of gestation. The dams and fetuses were examined on d 21 (rats) and 30 (rabbits) of gestation, using standard teratological methods. Indium concentration was determined in the maternal and fetal blood, as well as in the amniotic fluid, by atomic absorption spectrometry. Indium was found to cross the placenta and appeared in fetal blood in proportion to the metal concentration of the maternal blood. In the amniotic fluid, indium concentrations remained below the detection limit. In rats, indium chloride produced dose dependent maternal toxic effects, with a dose of 400 mg/kg inducing embryotoxicity (embryolethality) and teratogenicity. Doses of 200 and 100 mg/kg were embryotoxic (retarding) and teratogenic, causing skeletal and visceral anomalies in addition to external anomalies (rudimentary or missing tail, syndactylia, clubfoot, exencephalia) in rats. In rabbits, 200 mg/kg indium chloride was lethal for the dams and the embryos (some of the animals died, and the number of abortions and full resorptions increased). This dose was found to be teratogenic (caused gross renal anomalies) and increased the frequency of fetuses with skeletal retardation. In rats, the effects of indium chloride causing fetal retardation was found to be independent of exposure time. The teratogenic effects were the highest on d 11 and 12 of gestation, when indium chloride caused gross external malformations. Data suggest that the teratogenic effects of indium chloride can be attributed primarily to a direct cytotoxic action of indium resulting from placental transfer, but the effect is not a selective one, as it appears only in the presence of maternal toxic effects. PMID- 10681098 TI - Effect of cadmium on Pekin duck total body water, water flux, renal filtration, and salt gland function. AB - The following hypotheses were examined using Pekin ducks (Anas platyrhynchos) as a model for marine ducks: cadmium (Cd) intake affects (1) salt gland and/or kidney function of ducks and (2) osmoregulation differently in male and female ducks. Birds were fed 0, 50, or 300 microg Cd/g food. They were gradually acclimated to 450 mM NaCl and then drank 300 mM NaCl for 3 mo while salt gland secretion (SGS), glomerular filtration rate (GFR), total body water (TBW), and water flux (WF) were measured in ducks eating control and high-Cd diets. Cadmium ingestion did not markedly affect body mass, but significantly enlarged the salt glands and kidneys. Enhancement of kidney mass was greater in males. Cadmium ingestion did not affect TBW or WF, but tended to increase interstitial fluid space at the expense of intracellular fluid. Sex did not affect TBW, but males had greater WF. Birds that ate Cd diets, especially the higher Cd diet, exhibited renal tubular damage and lower GFR. Ducks that ate Cd had lower plasma sodium concentration and osmolality and, to activate SGS, required longer infusion of NaCl and larger increments PMID- 10681099 TI - Comparison of oxime-initiated reactivation of organophosphorous-inhibited acetylcholinesterase in brains of avian embryos. AB - Organophosphorous (OP) insecticide-induced inhibition and oxime reactivation of acetylcholinesterase (AChE) was determined in whole-brain homogenates prepared from 15-d-old chick embryos. Doses of chlorpyrifos, parathion, acephate, and trichlorfon that inhibited AChE >70% were administered to the embryos. Following insecticide exposure, an in vitro system compared the capability of the oximes pralidoxime (2-PAM), obidoxime, TMB-4, and HI-6 to reactivate the OP-inhibited AChE. Concentration-related increases in AChE activities were noted in embryo brains reactivated with 2-PAM, TMB, and HI-6. 2-PAM was the most effective reactivator of trichlorfon-inhibited AChE; 2-PAM and obidoxime were relatively similar in effectiveness for reactivation of AChE inhibited with the other OP insecticides used as test agents. All oximes were similarly effective against acephate, but HI-6 was the least effective reactivator of AChE in chick embryo brain homogenates inhibited by the other OP insecticides. These results suggest that both the OP insecticide inhibiting AChE and the oxime reactivating this enzyme can contribute to the effectiveness of the avian brain AChE reactivation. PMID- 10681100 TI - Effectiveness of comprehensive services for crack-dependent mothers with newborns and young children. AB - This article presents an outcome study of the Family Rehabilitation Program (FRP), a unique network of community-based programs in New York City that provides comprehensive services to families with drug-dependent parents, most caring for prenatally cocaine-exposed newborns. An admission sample of 173 mothers in 17 FRP sites was studied for one year; substance use was assessed by hair analysis and self-report. Mean length of retention was 10 months; half the clients were still active in the program at follow-up. Mothers completing or still active in FRP had higher rates of abstinence and substantially lower average levels of cocaine in their hair at follow-up than those exiting prematurely. The percent of families with children out of their homes did not increase significantly between admission and follow-up, and completing or remaining active in the program were associated with less out-of-home placement at follow-up. PMID- 10681101 TI - Residential treatment for women with dependent children: one agency's approach. AB - The Salvation Army First Choice Program, located in Fort Worth, Texas, provides comprehensive-as well as gender-specific-treatment for addicted women while providing child care and therapeutic services for children. Specific program attributes (including therapeutic interventions, community linkages, and staffing patterns) are described, and the five-year evaluation initiative, designed to examine relationships between client characteristics, program participation, and client progress is outlined. Findings from initial analyses examining correlates of 90-day dropout suggest a complex interaction among specific problems a woman brings to treatment, her level of dysfunction at treatment entry, how much social support is available to her, and what services she receives. PMID- 10681102 TI - Criminality among female drug abusers. AB - Criminality among female (n=351) drug abusers is compared to that of men (n=798) as part of a longitudinal study of persons in treatment in Sweden (the SWEDATE project). The extent of criminality was much less among females than among males, and fewer women than men were criminal. The pattern of criminality varied between the sexes. Women's crime debuts occurred later, and they committed less violent crimes and more drug-related crimes. The majority of women supported themselves in other ways than with criminality. Also, women tended to have a more severe pattern of abuse, a more rapid drug career, and more complex psychological problems than men. A subgroup of prostitutes whose drug of choice was heroin often began drug use early with cannabis and went on to amphetamine for their first injection, which often took place in a junkie pad. There was also a criminal group (as there was among men) with a very early and intensive juvenile delinquency pattern, early drug debuts and a rapid transition to regular abuse and extensive adult criminality. Forty-two percent of the women had no criminal records; they had more extensive multiple drug abuse than the other women (this was also true for the noncriminal male addicts). The study shows that drug abuse and criminality are interrelated for certain individuals, but not for others. PMID- 10681103 TI - Drug treatment outcomes: investigating the long-term effects of sexual and physical abuse histories. AB - Individuals in drug treatment, particularly women, generally report high levels of past sexual and physical abuse. Although histories of sexual and physical abuse are associated with greater prevalence and severity of depression, anxiety, phobias, and interpersonal difficulties for individuals seeking substance-related treatment, several recent studies failed to show that prior sexual or physical abuse compromised short-term drug treatment outcomes. This study examined the possible effects of sexual and physical abuse on a wide array of behavioral domains over a two-year posttreatment period. The findings indicate few differences between those with and without past histories of such abuse in terms of drug use, drug treatment and 12-Step program participation, criminality, income sources, intimate relationships, family functioning, and psychiatric symptoms. There are specific exceptions, but they apply only to men. Overall, the findings indicate that the impact of sexual and physical abuse histories on relatively long-term treatment outcomes is minimal. Addressing the sexual and physical abuse histories of those seeking treatment for drug abuse may be justified on humanistic grounds, but it will not significantly improve the long term effectiveness of drug treatment, nor will it substantially enhance the lives of those with histories of abuse. PMID- 10681104 TI - Using naltrexone in inpatient alcoholism treatment. AB - Naltrexone has been used successfully in outpatient settings as an adjunct to alcoholism treatment. This study examines the efficacy of using naltrexone in an inpatient treatment setting. Sixty-three alcohol-dependent patients who volunteered for a double-blind, placebo-controlled study were followed over the course of their 20 days in treatment and six months follow-up. A comparison group of 59 patients who did not volunteer were also studied over the same period of time. Patients in the study group were randomly assigned to receive naltrexone or placebo. Information was gathered daily on alcohol craving, drug craving and moods on self-reporting forms from the naltrexone and placebo groups, and from the comparison group. Follow-up data was gathered through self-report and through Washington State's TARGET management information system. No significant differences were found in craving scores while in treatment, nor in recidivism after treatment. PMID- 10681105 TI - Regional cerebral blood flow in alcohol-induced violence: a case study. AB - A case is presented of a 20-year-old man who became violent on many occasions after ingesting alcohol. On one occasion he committed an armed robbery. Two brain SPECT studies were performed: one when he was alcohol free, and one after he ingested alcohol in the same pattern as the night of the crime. The "alcohol free" study revealed marked hyperactivity in the cingulate gyrus, right and left lateral frontal lobes, right and left lateral parietal lobes and the right lateral temporal lobe. The "alcohol intoxication" study showed an overall dampening effect on the hyperactive areas of the brain, with only the anterior cingulate gyrus showing excessive activity. In addition, the right and left prefrontal cortex became hypoperfused, decreasing impulse control and judgment, as did the left and right temporal lobes, increasing the likelihood for aggression. This study suggests that this man may have been "self-medicating" an overactive brain, but in the process induced a state that increased the likelihood for aggressive behavior. This case study suggests the need for further research in the area of alcohol-induced violence and the potential usefulness of SPECT imaging, although no conclusions can be drawn from one case. PMID- 10681106 TI - A preliminary study of drug abuse and its mental health and health consequences among addicts in Greater Accra, Ghana. AB - This article represents a preliminary effort to describe drug abuse in Tudu, one of a number of neighborhoods in Accra that serve as drug centers. The problems of such neighborhoods reflect the drug problems that currently beset the rest of Ghana and Accra in particular. There is almost no fundamental current research on this issue. The few works cited comprise virtually the entire body of existing literature on this subject, and they fall far short of providing a comprehensive account of the changes that drugs have made in the social structure of the greater Accra region. This article is based on research done in the drug parlors and alleyways where the Tudu drug trade is conducted, and is a preliminary effort to redress the current lack of information by documenting the changing patterns of drug use in greater Accra. The findings reveal that a shift is underway from traditional marijuana abuse to abuse of crack cocaine and heroin. The article highlights the social relations that characterize this more dangerous drug setting and enhances the understanding of the psychiatric comorbidity of drug abuse, health, and behavior. These conclusions are derived from a multifaceted approach to data collection, taken to enhance the validity of research findings. PMID- 10681107 TI - Effects of control and motivation on treatment outcome. AB - This investigation was designed to determine who would benefit most from enhanced motivation, increased awareness of control, and the opportunity to make choices in substance abuse treatment. Thirty males and 21 females participated in treatment planning, goal setting, and determining methadone dosage level. Subjects were assigned at random to the Choice or Force condition. Choice group subjects were allowed to choose the type of treatment and their urinalysis schedule. No significant differences between groups on treatment outcome variables were found. However, higher motivation at the start of treatment predicted greater depression after 12 weeks. Results suggest that more highly motivated individuals seeking relief from substance dependency may be more vulnerable to depression which, in turn, can attenuate treatment effectiveness. PMID- 10681108 TI - A survey of adult recreational drug use via the World Wide Web: the DRUGNET study. AB - DRUGNET was a cross-sectional survey of adult recreational drug users (i.e., not abusers) via the World Wide Web of the Internet. The purpose of this survey was to provide a unique, broad description of nondeviant adult recreational drug users. The survey instrument had four divisions: demographic and lifestyle indices, drug use history, legal history and attitudes about drug issues, and the General Well-being Schedule (GWBS). Responses were received from 1,473 self identified drug users. Of these, 567 completed only the first section, leaving 906 respondents who completed the entire survey. The typical respondent was a White male who was well educated, employed full-time, a participant in recreational and community activities, and who described his physical health status as good. Their mental health, as measured by the GWBS, was similar to the general adult U.S. population. Their drug-taking behavior appeared to be well controlled, at mild to moderate levels in both frequency of use and degree of intoxication. These findings have major implications for drug policy and indicate the need for further research on the majority of drug users, who may be expected to resemble this sample more than they do clinical populations of drug abusers. PMID- 10681109 TI - The social construction of the crack epidemic in the print media. AB - Early news coverage of the rapid expansions and horrors associated with use of crack in the mid-1980s led to a great panic. Scholarly research subsequently debunked the various myths emanating from this media scare. This article examines whether this expanded understanding was reflected in the quality of news coverage over time through a comprehensive examination of all articles about crack cocaine appearing in the New York Times, Time, and Newsweek from 1985 through 1995. It was found that later news stories were generally less blatant, but overall did not correct for previous exaggerations. The long-term perspective also revealed an insidious bias in news coverage through its focus on the inner city, in spite of broader use of crack. This misleading view of "the problem" constructed by the media probably helped divert attention from persistent structural problems facing the inner-city. Scholars and activists need to continue their efforts to focus attention on the underlying social problems to counteract the media's propensity to focus on what might more appropriately be termed symptoms, such as the "crack epidemic." PMID- 10681110 TI - Teotlaqualli: the psychoactive food of the Aztec gods. AB - The Aztecs in pre-Columbian Mexico used not only a large number of single hallucinogens, they also used some combinations. The present article describes reports of the use of teotlaqualli, an unction prepared from ololiuhqui and picietl, with a large number of additions. The work of the chroniclers of pre Columbian Mexico served as a source of information. The teotlaqualli was offered to the gods, for whom it served as food. The Aztec priests smeared themselves with this unction, to lose fear and to get the appropriate state of mind to serve the Aztec gods. A few cases are reported in which the Aztec emperor or soldiers were smeared with teotlaqualli. It is suggested that the black color of some Aztec deities, as depicted in the codices, was due to anointment with teotlaqualli. In addition to its use for psychoactive purposes, teotlaqualli was used in medicine under the name teopatli. PMID- 10681111 TI - The status of psychologists' training about and treatment of substance-abusing clients. AB - A random survey of 1,200 psychologists (with a 62% response rate) indicated that most psychologists (91%) are to some degree involved in clinical practice with substance abusers, although most have no formal education (74%) or training (54%) in substance abuse. Relevant recommendations are made. PMID- 10681112 TI - Subjective appraisal of problem severity and the ASI: secondary data or second opinion? Addiction Severity Index. AB - The Addiction Severity Index is a popular research and clinical tool for the characterization of individuals grappling with substance abuse problems. For research, use of the seven objectively calculated composite scores of problem severity is recommended. In contrast, clinical use of the instrument relies more upon its subjectively derived interviewer and client severity rating scores. However, little systematic research has looked at the comparability of these two sources of client data. This study compared the objective and subjective scores of the ASI of male (n=141) and female (n=58) clients entering substance abuse treatment. In addition, clients' narratives about their most worrisome problems were recorded and put to content analyses. While significant correlations were found among the various subjective indices, little relationship could be discerned between the objective composite scores and any of the subjective indices derived from either the ASI or the clients' narratives. As the focus of outcome research shifts from objective client and treatment characteristics to a better understanding of the process of intervention, empirical characterization of substance abuse treatment outcome may be enriched by the inclusion of subjective data that taps into the client's own perceptions of problems and treatment efficacy in addition to more objective sources of data. PMID- 10681113 TI - Therapeutic use of cannabis by crack addicts in Brazil. AB - This study ensued from clinical observations based on spontaneous accounts by crack abusers undergoing their first psychiatric assessment, where they reported using cannabis in an attempt to ease their own withdrawal symptoms. Throughout a period of nine months, the researchers followed up on 25 male patients aged 16 to 28 who were strongly addicted to crack, as diagnosed through the Composite International Diagnostic Interview (CIDI), according to CID-10 and DSM-IV diagnostic criteria. Most of the subjects (68%, or 17 individuals) ceased to use crack and reported that the use of cannabis had reduced their craving symptoms, and produced subjective and concrete changes in their behavior, helping them to overcome crack addiction. The authors discuss some psychological, pharmacological and cultural aspects of these findings. PMID- 10681114 TI - Presentation of words to separate hemispheres prevents interword illusory conjunctions. AB - We tested the hypothesis that division of inputs between the hemispheres could prevent interword letter migrations in the form of illusory conjunctions. The task was to decide whether a centrally-presented consonant-vowel-consonant (CVC) target word matched one of four CVC words presented to a single hemisphere or divided between the hemispheres in a subsequent test display. During half of the target-absent trials, known as conjunction trials, letters from two separate words (e.g., "tag" and "cop") in the test display could be mistaken for a target word (e.g., "top"). For the other half of the target-absent trails, the test display did not match any target consonants (Experiment 1, N = 16) or it matched one target consonant (Experiment 2, N = 29), the latter constituting true "feature" trials. Bi- as compared to unihemispheric presentation significantly reduced the number of conjunction, but not feature, errors. Illusory conjunctions did not occur when the words were presented to separate hemispheres. PMID- 10681115 TI - Broad band spectral EEG parameters correlated with different IQ measurements. AB - The relationship of IQ (measured by WAIS and all its subscales) and EEG broad band spectral parameters were studied in 40 right-handed, male volunteers ranging in age from 20 to 25 years old. EEGs were recorded in 20 derivations during rest with eyes opened. The results obtained reveal positive and negative correlations with abundant frontal participation in all bands. Mean frequency data show a frequency shift in a very narrow range suggesting that more relations in narrow band could be achieved. These results highlight the need of psychological tests that measure more homogeneous abilities and finer measurement technique to reveal clearly explainable correlations and demonstrate that EEG recordings do reflect intellectual abilities. PMID- 10681116 TI - Spatial aspects of letter cancellation performance in Arabic readers. AB - Studies of visuospatial and directed attention that used subjects drawn from cultures with left-to-right reading patterns have suggested a slight performance bias toward left space. This pattern could reflect an intrinsic, organic, bias in spatial processing or the confounding effect of overlearned reading patterns. We studied the spatial distribution of errors on random array letter cancellation tasks obtained from 128 healthy Syrians who were native readers of Arabic. Fifty eight of the 128 subjects (45.3%) made a total of 91 errors in which they omitted cancelling a target. The distribution of errors was not spatially biased. This differs from the error pattern reported for native readers of English on a similar task. The findings, consistent with results of other approaches, suggest that reading patterns influence visuospatial attention, but are not the sole cause of spatial biases observed in readers of Indo-European languages. PMID- 10681117 TI - Patterns of waking EEG spectral power in chemically intolerant individuals during repeated chemical exposures. AB - Previous studies indicate that low level chemical intolerance (CI) is a symptom of several different controversial conditions with neuropsychiatric features, e.g., chronic fatigue syndrome, fibromyalgia, multiple chemical sensitivity, and "Persian Gulf Syndrome". Prior studies suggest that limbic and/or mesolimbic sensitization may contribute to development of CI. The purpose of this report was to document the waking electroencephalographic (EEG) patterns of individuals with CI during chemical exposures presented over repeated sessions. Three groups of adult subjects who were recruited from the community participated in the study: self-reported CI who had made associated lifestyle changes due to their intolerance (CI/ LSC), self-reported CI who had not made such changes (CI), and normal controls without self-reported CI. Subjects underwent two sessions involving one-minute EEG recordings during exposures to low level chemical odors (a probe for limbic activation). The CI, but not the CI/ LSC, subjects had increased absolute delta power after the chemical exposures during the second, but not the first, session. The findings support the neural sensitization hypothesis for intolerance to low levels of environmental chemicals in vulnerable individuals. As in human studies of stimulant drug sensitization, those with the strongest past history with sensitizing agents may not show-term sensitization to low level exposures in the laboratory. PMID- 10681118 TI - Expanding the neurological examination using functional neurologic assessment part I: methodological considerations. AB - Manual assessment of muscular function, in particular a method known as applied kinesiology (AK), is a clinical measure of neurologic function. A review of the literature reveals methodological problems with previous studies of AK as a form of neurologic assessment. Research designs that do not reflect clinical practice and principles of AK are common in the literature. Additional study is warranted to explore the potential of AK manual muscle testing as a diagnostic tool. We outline principles of AK and recommend that future research reflect more accurately the clinical practice of functional neurologic assessment and applied kinesiology. PMID- 10681119 TI - Expanding the neurological examination using functional neurologic assessment: part II neurologic basis of applied kinesiology. AB - Functional Neurologic Assessment and treatment methods common to the practice of applied kinesiology are presented. These methods are proposed to enhance neurological examination and treatment procedures toward more effective assessment and care of functional impairment. A neurologic model for these procedures is proposed. Manual assessment of muscular function is used to identify changes associated with facilitation and inhibition, in response to the introduction of sensory receptor-based stimuli. Muscle testing responses to sensory stimulation of known value are compared with usually predictable patterns based on known neuroanatomy and neurophysiology, guiding the clinician to an understanding of the functional status of the patient's nervous system. These assessment procedures are used in addition to other standard diagnostic measures to augment rather than replace the existing diagnostic armamentarium. The proper understanding of the neurophysiologic basis of muscle testing procedures will assist in the design of further investigations into applied kinesiology. Accordingly, the neurophysiologic basis and proposed mechanisms of these methods are reviewed. PMID- 10681120 TI - Visual evoked potentials and heart rate during white noise stimulation. AB - Visual evoked potentials (VEPs) were recorded in 12 adult participants as a function of the temporal frequency of a phase-reversed checkerboard, with or without a simultaneously presented white noise. During the VEP recordings also the pulse rate was measured. VEP amplitude changed as function of temporal frequency, but it was not affected by noise. Pulse rate was stable during the session without noise, but it increased during the white noise stimulation at high temporal frequencies. Heart acceleration might be associated to conditions when processing at low levels of visual sensitivity (high temporal frequencies) is furthermore disturbed by interfering stimulation (noise). PMID- 10681121 TI - Orchidectomy affects the innervation of the rat thymus. AB - To assess a putative role of the neural pathways in transfer of information from the gonads to the thymus, adult AO rats were orchidectomized (ORX) or sham ORX (controls); sacrificed 1, 3, 7, or 15 days later and their thymi were analyzed for: (a) the concentrations of noradrenaline (NA), dopamine (DA) and serotonin (5 HT) and distribution of monoamine-containing nerve profiles and (b) the acetylcholinesterase (AChE) activity and distribution of AChE-containing nerve profiles. Three days after the castration, an elevation in the level of both catecholamines, reflecting an increase in the overall intensity of nerve fibers autofluorescence, was found. Seven days post castration neither NA nor DA concentration differed from the appropriate control values, while 15 days after the surgery the concentration of NA was lower than that in the controls, most likely, due to diminished density of noradrenergic nerve profiles. In both the rats sacrificed 7 and 15 days after orchidectomy the concentration of 5-HT was reduced as result of a decrease in the density of 5-HT-containing autofluorescent cells. The activity of AChE was depressed one day after the surgery; then increased, so that 3 days post castration its value was higher than that in the sham ORX. After this increase, AChE activity decreased being, at postoperative day 7 and 15, lower than that in the controls. It seems that this decrease in AChE activity reflected, not only a reduction in the density of AChE-containing nerve fibers, but also a decrease in the density of AChE positive cells. Thus, the results indicate that orchidectomy can evoke changes in the T-cell maturation altering modulatory influences on this process coming via neural route, as well as those coming from the mast cells and AChE positive epithelial cells which constitute important component of the thymus microenvironment. PMID- 10681122 TI - Serotonergic neuronal sprouting as a potential mechanism of recovery in multiple sclerosis. AB - Experimental allergic encephalomyelitis (EAE) is widely considered as an animal model of multiple sclerosis (MS). Damage to the bulbospinal serotonergic (5-HT) neurons occurs in the early paralytic stages of EAE in rats with the severity of neurologic signs corresponding to spinal serotonergic depletion. Neurologic recovery of EAE rats is associated with reestablishment of spinal 5-HT transmission possibly through sprouting of undamaged axons and nerve terminals. Damage to the bulbospinal serotonergic fibers also occurs in patients with MS (as reflected by reduced lumbar CSF 5-HIAA levels) and may contribute to several manifestations of the disease including autonomic dysregulation, sensory symptoms (i.e., paresthesias, pain) and motor symptoms (weakness, spasticity, clonus). Spinal serotonergic neuronal sprouting with regeneration of 5-HT nerve terminals may also occur in the early stages of MS and may be associated with spontaneous remission of MS symptoms following an acute relapse. Sprouting of serotonergic neurons may also explain the disparity in MS between the extent of demyelinating plaques and clinical signs of the disease. The chronic course of MS may be associated with progressive axonal degenerative changes with reduction of serotonergic nerve terminals and loss of their sprouting capability. It is proposed that the beneficial effects of treatment with AC pulsed electromagnetic fields on the symptoms and course of the disease in patients with chronic progressive MS may be related in part to renewed sprouting of serotonergic neurons. PMID- 10681123 TI - Yawning and stretching induced by transcranial application of AC pulsed electromagnetic fields in Parkinson's disease. AB - Yawning is considered a brainstem regulated behavior which is associated with changes in arousal and activity levels. Yawning and stretching are dopamine (DA) mediated behaviors and pharmacological studies indicate that these behaviors are associated with increased DA release coupled with stimulation of postsynaptic DA D2 receptors. Despite their relation to the dopaminergic system, yawning and stretching are poorly documented in untreated or treated patients with Parkinson's disease (PD). A 49 year old fully medicated female patient with juvenile onset PD is presented in whom recurrent episodes of yawning and stretching developed during transcranial administration of AC pulsed electromagnetic fields (EM Fs) of picotesla flux density. These episodes have not been observed previously in this or other patients during treatment with levodopa or DA receptor agonists or in unmedicated PD patients during treatment with AC pulsed EMFs. It is suggested that yawning and stretching behavior resulted in this patient from a synergistic interaction between EMFs and DA derived from levodopa supplementation with EMFs possibly facilitating the release of DA and simultaneously activating postsynaptic DA-D2 receptors in the nigrostriatal dopaminergic pathways. In addition, it is postulated that the release of ACTH/MSH peptides from peptidergic neurons in the brain upon stimulation of the DA-D2 receptors reinforced the yawning and stretching behavior. PMID- 10681124 TI - It's not all in our genes! PMID- 10681125 TI - Paper Alert. Cell Biology. PMID- 10681126 TI - Web alert. Cell differentiation cell multiplication. PMID- 10681127 TI - Cell multiplication. Peering in and peering out: regulation of and by the cell cycle. PMID- 10681128 TI - Anaesthetic Research Society. Plymouth, July 8-9, 1999. Abstracts. PMID- 10681129 TI - [The appearance of the ophthalmoscope and the biomicroscope]. PMID- 10681130 TI - Results of mental health needs assessments performed by four urban American Indian organizations. PMID- 10681131 TI - 5-Hydroxy-4-oxo-L-norvaline depletes intracellular glutathione: a new modulator of drug resistance. AB - To search for compounds that reverse the drug resistance induced by glutathione (GSH), an original screening system to detect intracellular GSH depleters was established. Among 8843 microbes derived from the soil samples tested, the extracts of two Streptomyces species named KS6701 and KS8846, lowered the intracellular GSH level of Saccharomyces cerevisiae 5 x 47. From both the microbes, 5-hydroxy-4-oxo-L-norvaline (HON) was isolated as the active compound. At a concentration of 50-100 micrograms/ml, HON also decreased the GSH/protein level of the human ovarian tumor cell line, 2008/C13*5.25 and reversed its resistance to cisplatin. We also investigated the mechanism of the depletion. HON had little effect on gamma-glutamylcysteine synthetase (gamma-GCS) or glutathione synthetase, but HON decreased the quantity of thiol substances when it was spontaneously reacted with them. This suggested that the GSH depletion by HON occurred through a mechanism different from that of buthionine sulfoximine, a selective gamma-GCS inhibitor. PMID- 10681132 TI - Certification examination of the College of Family Physicians of Canada. Part 4: Simulated office orals. PMID- 10681133 TI - Acute otitis media. Evidence base on managing acute otitis media had inaccuracies. PMID- 10681134 TI - Acute otitis media. Surgery has a limited but important role in managing acute otitis media. PMID- 10681135 TI - Surgical treatments should have been discussed. PMID- 10681136 TI - Social and productive activities in elderly people. Activity (occupation) is important for survival. PMID- 10681137 TI - Social and productive activities in elderly people. Self rated health is important predictor of mortality. PMID- 10681138 TI - Randomised controlled trials in psychiatry. Scarcity of evidence is not necessarily evidence against long term psychodynamic psychotherapy. PMID- 10681139 TI - Evaluation of questionnaire on cancer family history on general practice. General practitioners reassure those at low risk and refer those at high risk. PMID- 10681141 TI - Significance of relationships. PMID- 10681140 TI - Bacteria associated with butterfly stain of Chilean tepa. AB - Butterfly stain, common in Chilean tepa (Laureliopsis philippiana [Looser] Schodde) trees, appears in cross-section of the stem as a series of partially overlapping orange-brown arcs that in early stages resembles a butterfly. Bacterial isolates from stained tepa wood samples cultured under aerobic and anaerobic conditions were identified using several commercially available identification systems. Pseudomonas sp. were most common in the cultures handled aerobically, and Clostridium sp. were found in the cultures handled anaerobically. A pectynolitic Bacillus sp. and a Gram-negative rod-shaped bacterium that emitted a strong naphthalene-like odor similar to that of stained wood were also isolated. Not all isolates could be identified. This the first report of Clostridium from tepa. PMID- 10681142 TI - Clinical expertise and the DPT: a need for residency training. PMID- 10681143 TI - Making jargon from kinetic and kinematic chains. PMID- 10681144 TI - Treatment of shoulder impingement syndrome. PMID- 10681145 TI - Studies of reliability: show me the question. PMID- 10681147 TI - Secrets of diagnosis. PMID- 10681146 TI - Grappling with the "miracle" of glucosamine. PMID- 10681148 TI - EIIaCre -- utility of a general deleter strain. PMID- 10681150 TI - Exposure-response relations of alpha-amylase sensitisation in British bakeries and flour mills. PMID- 10681152 TI - Gastrointestinal Endoscopy 2000. Conference proceedings. New York City, New York, USA. March 27-28, 1999. PMID- 10681151 TI - [The resistance of soil microorganisms to soil pollution by heavy metals]. AB - Microbial communities of grey podzolized soil of Left-Bank Ukraine are characterized by high potential resistance to pollution with heavy metals: above 40% of organotrophic microorganisms are capable to resist to pollution with a mixture of heavy metals (Cu2+, Cd2+, Pb2+, Zn2+) in a dose of 20 MPC of each metal. Relative amount of resistant microorganisms in polluted soil increases up to 93%. White zeolites and biohumus are promising for reclamation of soils polluted with heavy metals. They decrease toxic effect of heavy metals on soil microflora and promote development of microflora resistant to pollution. PMID- 10681153 TI - Brain in the twentieth century. PMID- 10681154 TI - Proceedings of the International Symposium on Vaccinology. Paris, France, 18-20 November 1998. PMID- 10681155 TI - [Report by the German Society of Gerontology and Geriatrics e. V. Protocol of the meeting of the ZEAG. (Central European Society of Gerontologic/Geriatric Societies) on 22 March 1999, 1 p.m., at the new Bad Hofgastein Congress Center]. PMID- 10681156 TI - [A trial of clonazepam treatment for manic-depressive psychoses]. AB - Clonazepam, which is a benzodiazepine structurally related to chlordiazepoxide hydrochloride, diazepam and nitrazepam, has been available for the treatment of seizure disorders in the USA since 1976 and in Japan since 1981. Recently, clonazepam has attracted interest as drug for the treatment of manic-depressive psychosis. Chouinard et al (1983), Victor et al (1984), Freinhar and Alvarez (1985) and Santos and Morton (1987) have reported on the antimanic properties of clonazepam, and Jones and Chouinard (1985), Zetine and Freedman (1986), Mas et al (1993) and Kishimoto et al(1987) have reported on the antidepressive properties of clonazepam. We have observed the antimanic and antidepressive properties of clonazepam as well. Clonazepam is a useful therapeutic adjunct and the onset of its effect is rapid. We cannot, however, advocate the use of only the therapeutic agent for amelioration of manic-depressive psychosis. Clonazepam is a useful supplement and has a diverse optimum daily dose for acute manic and depressive states. Clonazepam should, at the time, be considered only as an alternative treatment for patients nonresponsive to conventional therapy, and should be used at a dosage of 3-6 mg/day for depressive state and 10 mg/day for acute manic state in combination with other psychotropic drugs. PMID- 10681157 TI - Appearance of highly sensitive cells to L-type Ca antagonist in cultured cerebellar granule cells. AB - We studied intracellular Ca2+ ([Ca2+]i) kinetics within single cells using and image-analyzing system. We prepared a culture dish with a bottom made of quartz (non-fluorescent glass) for reducing the background fluorescence during [Ca2+]i measurement. Furthermore, we changed the concentration of the poly-L-lysine solution which had been used to make cells adhere to the dish bottom from the conventional 5 micrograms/ml to 15 micrograms/ml. This modification improved cell viability, prevented colony formation, and therefore, provided favorable cell samples for detection within single cells. Then, we investigated 25 mM KCl-evoked changes in the [Ca2+]i level of rat cerebellar granule cells. We found that sensitivity to KCl was high in some cells but low in others. In a study using the highly sensitive cells, nicardipine, nimodipine and nifedipine were active even at nanomolar or picomolar levels, although their affinities to the L-type Ca ion channel have been reported to act only at micromolar levels. PMID- 10681158 TI - [The influence of L-triiodothyronine on the action of desipramine on beta and serotonin 2A receptor, monoamines in the rat brain]. AB - The co-administration of thyroid hormone and antidepressants is often useful in the treatment of refractory depression. The aim of this study is to clarify the anti-depressive mechanisms of repeated co- administration of thyroid hormone (T3) and desipramine (DMI) in the rat brain. Forced swimming test, open-field test, monoamine contents, beta receptor, and serotonin (5HT)2A receptor were compared in four experimental groups (control, T3, DMI, DMI+T3). In the forced swimming test, the (DMI+T3) group showed a significantly decreased immobility time compared with that of the control group. The noradrenaline (NA) content in the prefrontal cortex was highest in the (DMI+T3) group. Although beta receptor was not altered in any brain region in the (DMI+T3) group, 5HT2A receptor was significantly decreased, and the dopamine (DA) turnover rate was significantly increased in the prefrontal cortex. This study suggests that the addition of T3 enhanced the action of DMI alone on the monoaminergic system in the prefrontal cortex. PMID- 10681159 TI - [The vertebrate nervous system comprises an enormous number of cell types]. AB - Neurogenesis is regulated by basic-helix-loop-helix (bHLH) transcription factors in both vertebrates and insects. MASH1 and neurogenin, members of the bHLH genes, are expressed in subsets of neural precursors and control their differentiation. Downstream of the bHLH genes, PHD1, Phox2, and DRG11, which belong to PHD family genes, are expressed in specific lineages of neurons and are involved in their neuronal identity. Differentiated sensory and motor neurons express PEA3 and ER81, members, of the ETS family, which specify neuronal connectivity. During neuronal differentiation, these three families of transcription factors control developmentally distinct operations. Increasing the numbers of members of the families may underlie the generation of neuronal diversity. PMID- 10681160 TI - [Brain cytokine network and novel characteristics of microglia]. AB - Microglia, macrophage-like cells in the central nervous system (CNS), are multi functional cells; they play an important role in the removal of dead cells or their remnants by phagocytosis in CNS degeneration and are one of the important cells in the CNS cytokine network to produce and respond to a variety of cytokines. Although little is known about microglia in the normal CNS, it is obvious that they are quickly activated in all acute pathological events including apoptosis, neurodegeneration and inflammation. Activation of microglia in apoptosis is a double-edged response; under severe apoptotic conditions, microglia act as scavengers removing tissue debris and inducing apoptosis in damaged cells, whereas in more subtle injury they exert a surveillance function and might play a protective role. The transformation of resting microglia into full-blown phagocytes is strictly regulated. To understand the molecular basis of controlling mechanisms of microglia in apoptosis, the study requires in vivo models. For such purpose, we developed the brain-targeting gene delivery system using immortalized microglia, which can facilitate investigation into the roles of particular microglial genes in apoptosis and the gene therapy of several brain disorders. PMID- 10681161 TI - [Synapse-specificity and gene regulation during long-term potentiation]. PMID- 10681162 TI - Proceedings of the Cuban Biotechnology Conference, Havana, 1998. PMID- 10681163 TI - Commentary on future allergy practice article. PMID- 10681164 TI - Prayer in psychotherapy. PMID- 10681165 TI - Report from the United States of America. PMID- 10681166 TI - The eyes have it. PMID- 10681167 TI - 3rd Annual North American Program on Computer Assisted Orthopaedic Surgery (CAOS/USA '99). Pittsburgh, Pennsylvania, USA. June 7-19, 1999. Abstracts. PMID- 10681169 TI - Future Trends in the Management of Rheumatoid Arthritis. Proceedings of the International Rheumatology Round Tables, March 27-28, 1998 and February 19-20, 1999. PMID- 10681168 TI - Proceedings of the symposium on transport and trafficking in the malaria-infected erythrocyte. London, United Kingdom, 26-28 January 1999. PMID- 10681170 TI - Dermatology blues--Society of Investigative Dermatology 60th Annual Meeting, 5-9 May 1999, Chicago, USA. PMID- 10681171 TI - Hydroxyurea in psoriasis. PMID- 10681172 TI - Patient information sheet used at St John's Institute of Dermatology. HYDROXYUREA (Hydrea) Information Leaflet. PMID- 10681173 TI - A sporadic form of hypertrichosis cubiti. PMID- 10681174 TI - Pulsed dye laser treatment of subacute cutaneous lupus erythematosus. PMID- 10681175 TI - Erosive pustular dermatosis of the scalp following cryotherapy and topical tretinoin for actinic keratoses. PMID- 10681176 TI - Jessner's lymphocytic infiltrate treated with auranofin. PMID- 10681177 TI - Ventilation scintigraphy for the detection of bronchopleural fistulae. PMID- 10681178 TI - Simple design for rapid self-injection ictal SPET during aura. PMID- 10681179 TI - An investigation into the electro-physical characteristics of microbial cells during the metabolism of toxic compounds under conditions of limited O2 availability. AB - The purpose of the work reported here was to experimentally clarify the interconnection between changes in the electro-physical characteristics of microbial suspensions and processes of the metabolism of certain toxic compounds (acrylamide and p-nitrophenol (PNP)) in cells containing enzyme systems of the initial metabolism of these compounds. In this work, we used cells of two strains, Acinetobacter calcoaceticum A-122 and Brevibacterium sp. 13PA, which are capable of utilising PNP and acrylamide, respectively, as the sole source of carbon. Suspensions of these cells exhibited appreciable decreases in the magnitude of the electro-optical signal when the microbial metabolism of PNP and acrylamide was inhibited under conditions of limited O2 supply. This attests to a relationship between the electro-physical characteristics of microbial suspensions and cellular metabolic reactions and also to the negligibly small effect that the non-specific interaction of substrate with the cells has on the suspensions' electro-optical properties. PMID- 10681180 TI - The 1999 Thomas Hunt Morgan Medal. Salome G. Waelsch. PMID- 10681181 TI - The 1999 Genetics Society of America Medal. Charles H. Langley. PMID- 10681182 TI - The yeast HSM3 gene is not involved in DNA mismatch repair in rapidly dividing cells. PMID- 10681183 TI - The yeast HSM3 gene is involved in DNA mismatch repair in slowly dividing cells. PMID- 10681184 TI - The 1999 George W. Beadle Medal. Michael Ashburner. PMID- 10681185 TI - Epidemiology supports oral contraceptives as a risk factor in Crohn's disease. PMID- 10681186 TI - The role of psychological and biological factors in postinfective gut dysfunction. PMID- 10681187 TI - Is isolated idiopathic pancreatitis associated with CFTR mutations? PMID- 10681188 TI - Did prostaglandin E(2) stimulate glucose absorption in rat intestine? PMID- 10681189 TI - Abstracts presented for the 31st annual meeting of the Society of Gynecologic Oncologists. San Diego, California, February 5-9, 2000. PMID- 10681190 TI - [Hypothesis--Origin of the parietal cells]. PMID- 10681191 TI - Diversion proctocolitis with granulomatous vasculitis in a patient without inflammatory bowel disease. PMID- 10681192 TI - Sarcomatous transformation of chromophobe cell renal carcinoma. PMID- 10681193 TI - Vitamin A induced stellate cell hyperplasia and fibrosis in renal failure. PMID- 10681194 TI - Mullerian adenosarcoma of the uterus in association with tamoxifen therapy. PMID- 10681195 TI - Metastasizing hyalinizing spindle cell tumour with giant rosettes: report of a case with long survival. PMID- 10681196 TI - Exocyclic DNA Adducts in Mutagenesis and Carcinogenesis. Proceedings of the 2nd international conference. Heidelberg, Germany, September 1998. PMID- 10681197 TI - [Health economics: notes for a research agenda]. PMID- 10681198 TI - [The dreams and realities of the British National Health Service. Primary care as a gateway to health services]. PMID- 10681199 TI - [The gatekeeper role of the general practitioner and the reform of the National Health Service as seen from Barcelona, Spain]. PMID- 10681200 TI - [Toxic peripheral polyneuropathy outbreak in a labour environment]. PMID- 10681201 TI - [Use of nonparametric methods for controlling unseen confounding variables in ecological time-series studies]. PMID- 10681202 TI - [Heinz Walther (1919-1999)]. PMID- 10681203 TI - [Professor Bernd-Rudiger Balda on his 60th birthday]. PMID- 10681204 TI - [Propionibacterium acnes--pathogenetic significance in sarcoidosis?]. PMID- 10681206 TI - Employment, starting salaries and educational indebtedness of 1999 graduates of US veterinary medical colleges. PMID- 10681207 TI - Is a large number of sputum specimens necessary for the bacteriological diagnosis of tuberculosis? PMID- 10681208 TI - Detection of Bordetella holmesii using Bordetella pertussis IS481 PCR assay. PMID- 10681209 TI - Two Actinobacillus pleuropneumoniae serotype 8 reference strains in circulation. PMID- 10681210 TI - Acute renal failure in an infant associated with cytotoxic Aeromonas sobria isolated from patient's stool and from aquarium water as suspected source of infection. PMID- 10681211 TI - Rapid mini-preparation of fungal DNA for PCR. PMID- 10681213 TI - Designs for life: the science of biomechanics. A tribute to Professor R. McNeill Alexander FRS. PMID- 10681212 TI - Obtaining unacceptable results in assays for quantitation of human immunodeficiency virus type 1 RNA in plasma samples. PMID- 10681214 TI - Attentional blink for color. AB - Colored letters were presented by means of rapid serial visual presentation (RSVP). Two letters (T1 and T2) were targets that required a speeded (T1) or delayed (T2) response. T1 was shown on half of the trials. Two tasks were performed in the experimental conditions. Task 1 required a discrimination between 2 letters (when T1 was shown) or an indication that T1 was absent. In Experiment 1, Task 2 was to detect the presence of a unique color in the RSVP stream (e.g., green in a stream of alternating red and gray). In Experiment 2, Task 2 was to report the color of the first colored letter to appear in the RSVP stream. The lag between T1 and T2 in the RSVP stream was manipulated. Accuracy of color reports in Task 2 was poor at shorter lags and improved as the lag was lengthened in both experiments, demonstrating an attentional blink for chromatic information. Theoretical implications of the results are discussed. PMID- 10681215 TI - On the mitochondrial aspect of reactive oxygen species action in external magnetic fields. AB - Some retinoids or porphyrins can form radical pairs, alter transfer of electrons, and increase synthesis of reactive oxygen species in the mitochondrial respiratory chain. This leads to cell damage and apoptosis. We propose that co application of an external static magnetic field of several to several hundreds miliTesla (mT) can enhance those effects by further alteration of electron flow in the mitochondrial respiratory chain, facilitation of forbidden transitions from the triplet to the singlet state of retinoid or porphyrin radicals, and modification of radical pair amount. Since electron flow in the mitochondrial respiratory chain seems to possess fractal dynamics, the magnetic field can initiate self- organization of the flow into more regular patterns, and create optimal conditions for damage of cancer cells without any detriment for the normal counterparts. External low magnetic field should improve effectiveness and selectivity of retinoid chemoprevention or porphyrin photodynamic therapy. PMID- 10681216 TI - The case in support of Gh therapy. PMID- 10681217 TI - Vitamin A supplementation for extremely-low-birth-weight infants. PMID- 10681218 TI - Cytomegalovirus infection and HIV-I disease progression in infants born to HIV-I infected women. PMID- 10681220 TI - Mississippi is blessed by HMO failure. PMID- 10681219 TI - Autism and measles, mumps, and rubella vaccine: No epidemiological evidence for a causal association. PMID- 10681221 TI - Global epidemic of cardiovascular disease predicted. PMID- 10681222 TI - Ochre suppressor transfer RNA restored dystrophin expression in mdx mice. PMID- 10681223 TI - Reflections in mutation research. Reflections of Zhores Medvedev. PMID- 10681224 TI - Transmyocardial laser revascularization. PMID- 10681225 TI - Transmyocardial laser revascularization. PMID- 10681226 TI - Mitochondrial disease in patients with exercise intolerance. PMID- 10681227 TI - Mitochondrial disease in patients with exercise intolerance. PMID- 10681228 TI - Virostatic therapy for advanced lymphoproliferation associated with the Epstein Barr virus in an HIV-infected patient. PMID- 10681229 TI - Nephropathy in type 2 diabetes. PMID- 10681230 TI - Nephropathy in type 2 diabetes. PMID- 10681231 TI - Nephropathy in type 2 diabetes. PMID- 10681232 TI - Hypereosinophilic syndrome. PMID- 10681233 TI - Hypereosinophilic syndrome. PMID- 10681234 TI - An angiosarcoma in the left atrium. PMID- 10681235 TI - Diversity convention in the balance. PMID- 10681236 TI - Atmospheric chemistry. Better budgets for methyl halides. PMID- 10681237 TI - Robert A. Swanson (1947-99) PMID- 10681238 TI - [XXI National Congress of the Italian Society of Vascular Angiology and Pathology. Bologna, 28 November-1 December 1999. Proceedings]. PMID- 10681239 TI - [Use of antithrombotic drugs in the treatment of peripheral occlusive arterial diseases]. PMID- 10681240 TI - Risk factors for back pain incidence in industry: a prospective study. PMID- 10681241 TI - The bee venom test: comparisons with the formalin test with injection of different venoms. PMID- 10681242 TI - Hemisensory impairment in patients with complex regional pain syndrome. PMID- 10681243 TI - Resolution of psychological distress of whiplash patients following treatment by radiofrequency neurotomy: a randomized, double-blind, placebo-controlled trial. PMID- 10681244 TI - Absence of evidence is not evidence of absence (again) PMID- 10681245 TI - Workshop on parenting methods to minimize disruptive behavior. PMID- 10681246 TI - Workshop on practical and cost-effective issues of behavior management. PMID- 10681247 TI - Workshop on choosing the setting and medication for the difficult child. PMID- 10681248 TI - Evolution of the genetic covariance between male and female components of mate recognition: an experimental test. AB - The evolution of a positive genetic correlation between male and female components of mate recognition systems will result as a consequence of assortative mating and, in particular, is central to a number of theories of sexual selection. Although the existence of such genetic correlations has been investigated in a number of taxa, it has yet to be shown that such correlations evolve and whether they may evolve as rapidly as suggested by sexual selection models. In this study, I used a hybridization experiment to disrupt natural mate recognition systems and then observed the subsequent evolutionary dynamics of the genetic correlation between male and female components for 56 generations in hybrids between Drosophila serrata and Drosophila birchii. The genetic correlation between male and female components evolved from 0.388 at generation 5 to 1.017 at generation 37 and then declined to -0.040 after a further 19 generations. These results indicated that the genetic basis of the mate recognition system in the hybrid populations evolved rapidly. The initial rapid increase in the genetic correlation was consistent with the classic assumption that male and female components will coevolve under sexual selection. The subsequent decline in genetic correlation may be attributable to the fixation of major genes, or, alternatively, may be a result of a cyclic evolutionary change in mate recognition. PMID- 10681249 TI - Phylogenetic approaches to nomenclature: a comparison based on a nemertean case study. AB - Phylogenetic approaches to biological nomenclature are becoming increasingly common. Here I compare the behaviour of two such approaches, the phylogenetic system of definition and the phylogenetic system of reference, when there is a shift in the preference of phylogenetic hypotheses. The comparison is based on a case study from nemertean systematics and is the first to compare two different phylogenetic approaches throughout three stages of change, including two stages of phylogenetic nomenclature. It is concluded that a phylogenetic system of reference in combination with uninomials is superior in conveying phylogenetic information. PMID- 10681251 TI - [XLI Congress of the French National Society of Internal Medicine. Toulouse, 9-11 December 1999. Proceedings]. PMID- 10681250 TI - Total serum IgE and IgA antibody levels in healthy dogs of different breeds and exposed to different environments. AB - Total serum immunoglobulin (Ig) E and A levels were analysed in 233 healthy dogs as basis for comparison with atopic dogs in future studies. They were measured by ELISA in a group of non- colonised dogs of various breeds (group A) and three groups of colonised dogs including one German Shepherd and two Beagle kennels (groups B-D). IgE levels from non-colonised dogs were significantly higher than the ones of German Shepherds and Beagles C (P<0.05). IgA levels were alike in all groups except for the German Shepherds which displayed the lowest levels. Age and sex were not identified as common significant cofactors for IgE and IgA levels in all groups and IgE levels correlated negatively with IgA only in non-colonised dogs. In conclusion, IgE and IgA levels seem to be mainly influenced by genetic background. Thus use of total serum IgE as a diagnostic tool in the atopic dogs required extensive family data and therefore appears most suitable for research purposes within specific, well defined dog populations. PMID- 10681252 TI - [The president's look back]. PMID- 10681253 TI - [Quality guidelines: are they applicable in the private office?]. PMID- 10681254 TI - [The debate for an incisor. The multidisciplinary approach in dental injuries. The 12th Meeting of the Association of Former Students of the School of Dentistry of Geneva, 30 September and 1 October 1999]. PMID- 10681255 TI - [Anonymous: a majolica plate representing Saint Apollonia. The BonaDent Collection]. PMID- 10681256 TI - [The juridical aspects of quality management (quality assurance) in dentistry]. PMID- 10681257 TI - [Everything about intraoral video systems--part I. Advice on the purchase decision and on use]. PMID- 10681258 TI - [Dental Health Assistance Switzerland]. PMID- 10681259 TI - [Periodontal risks in daily practice--the initiation of guided diagnosis. The 1999 congress of the SSP (Swiss Society for Periodontics) in Lugano]. PMID- 10681260 TI - [Prosthodontics on the threshold into the year 2000, the determination of its status and the future prospects for dental practice. A report on the 2nd Easter symposium of 10 April-17 April 1999 in Mallorca]. PMID- 10681261 TI - [Active specific immunotherapy (ASI) in colon cancer]. PMID- 10681262 TI - Risk factors for death from asthma. PMID- 10681263 TI - Nebulised taurolidine and B cepacia bronchiectasis. PMID- 10681264 TI - Effect of salmeterol on airway eosinophils. PMID- 10681265 TI - Familial coagulation factor V deficiency caused by a novel 4 base pair insertion in the factor V gene: factor V Stanford [erratum]. PMID- 10681266 TI - Ladislav Tauc (1926-1999) PMID- 10681267 TI - [IV Conference on plant karyology and karyosystematics. St. Petersburg, May 25 27, 1999. Abstracts]. PMID- 10681268 TI - [Abstracts of papers and communications submitted to the International conference "Free-radical processes: ecological, pharmacological and clinical aspects". St. Petersburg, September 8-10, 1999]. PMID- 10681269 TI - B vitamins, homocysteine, and neurocognitive function in the elderly. AB - Evidence of the importance of the B vitamins folic acid, vitamin B-12, and vitamin B-6 for the well-being and normal function of the brain derives from data showing neurologic and psychologic dysfunction in vitamin deficiency states and in cases of congenital defects of one-carbon metabolism. The status of these vitamins is frequently inadequate in the elderly and recent studies have shown associations between loss of cognitive function or Alzheimer disease and inadequate B vitamin status. The question that arises is whether these B vitamin inadequacies contribute to such brain malfunctions or result from aging and disease. From a theoretical standpoint, these inadequacies could give rise to impairment of methylation reactions that are crucial to the health of brain tissue. In addition or perhaps instead, these inadequacies could result in hyperhomocysteinemia, a recently identified risk factor for occlusive vascular disease, stroke, and thrombosis, any of which may result in brain ischemia. Advances in the understanding of this putative relation between inadequate vitamin status and loss of cognitive function in the elderly are likely to be slow and may depend on the outcomes of both prospective studies and longitudinal studies in which nutritional intervention is provided before cognitive decline occurs. PMID- 10681270 TI - Oxidative stress and Alzheimer disease. AB - Research in the field of molecular biology has helped to provide a better understanding of both the cascade of biochemical events that occurs with Alzheimer disease (AD) and the heterogeneous nature of the disease. One hypothesis that accounts for both the heterogeneous nature of AD and the fact that aging is the most obvious risk factor is that free radicals are involved. The probability of this involvement is supported by the fact that neurons are extremely sensitive to attacks by destructive free radicals. Furthermore, lesions are present in the brains of AD patients that are typically associated with attacks by free radicals (eg, damage to DNA, protein oxidation, lipid peroxidation, and advanced glycosylation end products), and metals (eg, iron, copper, zinc, and aluminum) are present that have catalytic activity that produce free radicals. beta-Amyloid is aggregated and produces more free radicals in the presence of free radicals; beta-amyloid toxicity is eliminated by free radical scavengers. Apolipoprotein E is subject to attacks by free radicals, and apolipoprotein E peroxidation has been correlated with AD. In contrast, apolipoprotein E can act as a free radical scavenger and this behavior is isoform dependent. AD has been linked to mitochondrial anomalies affecting cytochrome-c oxidase, and these anomalies may contribute to the abnormal production of free radicals. Finally, many free radical scavengers (eg, vitamin E, selegeline, and Ginkgo biloba extract EGb 761) have produced promising results in relation to AD, as has desferrioxamine-an iron-chelating agent-and antiinflammatory drugs and estrogens, which also have an antioxidant effect. PMID- 10681271 TI - Vitamin E and Alzheimer disease: the basis for additional clinical trials. AB - Many lines of evidence suggest that oxidative stress is important in the pathogenesis of Alzheimer disease. In particular, beta-amyloid, which is found abundantly in the brains of Alzheimer disease patients, is toxic in neuronal cell cultures through a mechanism involving free radicals. Vitamin E prevents the oxidative damage induced by beta-amyloid in cell culture and delays memory deficits in animal models. A placebo-controlled, clinical trial of vitamin E in patients with moderately advanced Alzheimer disease was conducted by the Alzheimer's Disease Cooperative Study. Subjects in the vitamin E group were treated with 2000 IU (1342 alpha-tocopherol equivalents) vitamin E/d. The results indicated that vitamin E may slow functional deterioration leading to nursing home placement. A new clinical trial is planned that will examine whether vitamin E can delay or prevent a clinical diagnosis of Alzheimer disease in elderly persons with mild cognitive impairment. PMID- 10681272 TI - Weight loss in Alzheimer disease. AB - BACKGROUND: Epidemiologic studies have shown that weight loss is commonly associated with Alzheimer disease (AD) and is a manifestation of the disease itself. The etiology of weight loss in AD appears multifactorial. Hypotheses to explain the weight loss have been suggested (eg, atrophy of the mesial temporal cortex, biological disturbances, and higher energy expenditure); however, none have been proven. OBJECTIVE: In the first part of this article, we describe weight loss in AD (epidemiologic data and hypotheses to explain weight loss and anorexia in AD). In the second part we report the results of a longitudinal study of the changes in nutritional variables in a cohort of patients with a probable diagnosis of AD. DESIGN: We followed subjects with AD (based on criteria of the National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer's Disease and Related Disorders Association) who were recruited from the Alzheimer's Disease Center in Toulouse. All subject underwent a nutritional, neuropsychologic, and functional evaluation. The Zarit scales were used to assess caregiver burden and caregiver reactions to the patients' behavioral and autonomic disorders. RESULTS: We showed that only results of the Burden Interview and the Memory and Behavior Problems Checklist, which explored caregiver burden, predicted weight loss in AD. It is possible that caregivers who consider themselves overburdened by the disease process are not willing to invest adequate resources to allow AD patients to properly nourish themselves. CONCLUSION: Nutritional education programs for the caregivers of AD patients seem to be the best way to prevent weight loss and improve the nutritional status of these patients. PMID- 10681273 TI - Alzheimer disease: protective factors. AB - Approximately 6-8% of all persons aged >65 y have Alzheimer disease and the prevalence of the disease is increasing. Any intervention strategy aimed at decreasing risks or delaying the onset of the disease will therefore have a substantial effect on health care costs. Nutrition seems to be one of the factors that may play a protective role in Alzheimer disease. Many studies suggest that oxidative stress and the accumulation of free radicals are involved in the pathophysiology of the disease. Several studies have shown the existence of a correlation between cognitive skills and the serum concentrations of folate, vitamin B-12, vitamin B-6, and, more recently, homocysteine. However, nutritional factors have to be studied not alone but with the other factors related to Alzheimer disease: genetics, estrogen, antiinflammatory drug use, and socioeconomic variables. The objective of this article was to review recent studies in this field. PMID- 10681275 TI - Changes in definitions of clinical staging for carcinoma of the cervix and ovary: International Federation of Gynecology and Obstetrics. PMID- 10681276 TI - Proceedings of an International Symposium on the Mechanisms and Management of Allergic Disease to mark the 30th Anniversary of Allergopharma Joachim Ganzer KG. Hamburg, Germany, September 24-26, 1999. PMID- 10681274 TI - Energy expenditure, energy intake, and weight loss in Alzheimer disease. AB - Alzheimer disease is one of the leading causes of death among older individuals. Unexplained weight loss and cachexia are frequent clinical findings in patients with Alzheimer disease. Thus, it has been postulated that Alzheimer disease may be associated with dysfunction in body weight regulation. This brief review examines the interrelations among energy intake, energy expenditure, and body composition in Alzheimer disease. We explored whether abnormally high daily energy expenditures, low energy intakes, or both contribute to unexplained weight loss and a decline in nutritional status. Specifically, we considered studies that examined energy intake, body composition, and daily energy expenditure and its components. The application of doubly labeled water and indirect calorimetry to understand the etiology of wasting has increased our knowledge regarding the relation among energy expenditure, physical activity levels, and body composition in Alzheimer disease patients. Although the number of studies are limited, results do not support the notion that a hypermetabolic state contributes to unexplained weight loss in Alzheimer disease, even in cachectic patients. Recent findings are presented suggesting an association between abnormally elevated levels of physical activity energy expenditure and elevated appendicular skeletal muscle mass and energy intake in Alzheimer disease patients. Clinical strategies aimed at developing lifestyle and dietary interventions to maintain adequate energy intake, restore energy balance, and maintain skeletal muscle mass should be a future area of investigation in Alzheimer disease research. PMID- 10681277 TI - Opioid-induced breathing patterns disappear from view. PMID- 10681278 TI - Profound motor blockade with epidural ropivacaine. PMID- 10681279 TI - Effects of infant birthweight and maternal body mass index in pregnancy on components of the insulin resistance syndrome in China. AB - BACKGROUND: Reduced birthweight is associated with increased risk for the insulin resistance syndrome. Part of this risk is hypothesized to originate from undernutrition in utero. The prevalence of the insulin resistance syndrome increases in countries that undergo the transition from chronic malnutrition to adequate nutrition, when postnatal nutrition improves more rapidly than prenatal nutrition. OBJECTIVE: To determine whether the components of the insulin resistance syndrome are associated with reduced fetal growth and maternal undernutrition. DESIGN: A nonconcurrent, prospective study of men and women whose mothers' heights and weights were recorded during pregnancy. SETTING: Beijing, China. PARTICIPANTS: 627 men and women (mean age, 45 years) whose mothers' obstetric records were preserved. MEASUREMENTS: Adult offspring's blood pressure, plasma glucose levels, insulin levels, and lipid concentrations during an oral glucose tolerance test. The main explanatory measurements were mothers' body mass index during pregnancy and offspring's birthweight and adult size. RESULTS: After adjustment for sex and current body mass index, low birthweight was associated with elevated plasma glucose levels, insulin levels, triglyceride concentrations, and blood pressure. For every 1-kg increase in birthweight, systolic blood pressure decreased by 2.9 mm Hg (95% CI, 0.3 to 5.4 mm Hg) and the 2-hour plasma glucose level decreased by 5.1% (CI, 0.7% to 9.3%). Low maternal body mass index in early and late pregnancy was associated with elevated levels of plasma glucose, insulin, and triglycerides in adult offspring but was not associated with elevated blood pressure. CONCLUSIONS: Risk for the insulin resistance syndrome may be partially established through low maternal body mass before pregnancy and consequent fetal undernutrition. This risk is independent of that associated with adult obesity. In developing countries such as China, improved nutrition in girls and young women may offer long-term benefits to offspring. PMID- 10681280 TI - Screening for hereditary hemochromatosis in siblings and children of affected patients. A cost-effectiveness analysis. AB - BACKGROUND: Screening for hereditary hemochromatosis is traditionally done by using serum iron studies. However, mutation analysis of the hemochromatosis associated HFE gene has recently become available. OBJECTIVE: To compare the cost effectiveness of no screening with four screening strategies that incorporate HFE gene testing or serum iron studies. DESIGN: Cost-effectiveness analysis. DATA SOURCES: Published literature. TARGET POPULATION: Siblings and children of an affected proband. TIME HORIZON: Lifetime from 10 years of age (children) or 45 years of age (siblings). PERSPECTIVE: Societal. INTERVENTION: 1) Serum iron studies. 2) Gene testing of the proband. If the proband is homozygous (C82Y+/+), the spouse undergoes gene testing; if he or she is heterozygous (C82Y+/-), the children undergo gene testing. 3) Gene testing of the proband; if he or she is homozygous, relatives undergo gene testing. 4) Direct gene testing of relatives. OUTCOME MEASURES: Cost per life-year saved and incremental cost-effectiveness ratio. RESULTS OF BASE-CASE ANALYSIS: In children, HFE gene testing of the proband was the most cost-effective strategy for screening one child (incremental cost-effectiveness ratio, $508 per life-year saved). HFE gene testing of the proband followed by testing of the spouse was the most cost-effective strategy for screening two or more children (incremental cost-effectiveness ratio, $3665 per life-year saved). In siblings, all screening strategies were dominant compared with no screening. Strategies using HFE gene testing were less costly than serum iron studies. RESULTS OF SENSITIVITY ANALYSIS: Despite varying the prevalence of mutations and regardless of the cost of the genetic test in one- and two-way sensitivity analyses, HFE gene testing remained cost-effective. CONCLUSIONS: HFE gene testing for the C282Y mutation is a cost-effective method of screening relatives of patients with hereditary hemochromatosis. PMID- 10681281 TI - Subclinical hypothyroidism is an independent risk factor for atherosclerosis and myocardial infarction in elderly women: the Rotterdam Study. AB - BACKGROUND: Overt hypothyroidism has been found to be associated with cardiovascular disease. Whether subclinical hypothyroidism and thyroid autoimmunity are also risk factors for cardiovascular disease is controversial. OBJECTIVE: To investigate whether subclinical hypothyroidism and thyroid autoimmunity are associated with aortic atherosclerosis and myocardial infarction in postmenopausal women. DESIGN: Population-based cross-sectional study. SETTING: A district of Rotterdam, The Netherlands. PARTICIPANTS: Random sample of 1149 women (mean age +/- SD, 69.0 +/- 7.5 years) participating in the Rotterdam Study. MEASUREMENTS: Data on thyroid status, aortic atherosclerosis, and history of myocardial infarction were obtained at baseline. Subclinical hypothyroidism was defined as an elevated thyroid-stimulating hormone level (>4.0 mU/L) and a normal serum free thyroxine level (11 to 25 pmol/L [0.9 to 1.9 ng/dL]). In tests for antibodies to thyroid peroxidase, a serum level greater than 10 IU/mL was considered a positive result. RESULTS: Subclinical hypothyroidism was present in 10.8% of participants and was associated with a greater age-adjusted prevalence of aortic atherosclerosis (odds ratio, 1.7 [95% CI, 1.1 to 2.6]) and myocardial infarction (odds ratio, 2.3 [CI, 1.3 to 4.0]). Additional adjustment for body mass index, total and high-density lipoprotein cholesterol level, blood pressure, and smoking status, as well as exclusion of women who took beta-blockers, did not affect these estimates. Associations were slightly stronger in women who had subclinical hypothyroidism and antibodies to thyroid peroxidase (odds ratio for aortic atherosclerosis, 1.9 [CI, 1.1 to 3.6]; odds ratio for myocardial infarction, 3.1 [CI, 1.5 to 6.3]). No association was found between thyroid autoimmunity itself and cardiovascular disease. The population attributable risk percentage for subclinical hypothyroidism associated with myocardial infarction was within the range of that for known major risk factors for cardiovascular disease. CONCLUSION: Subclinical hypothyroidism is a strong indicator of risk for atherosclerosis and myocardial infarction in elderly women. PMID- 10681282 TI - Positive results on tests for steatorrhea in persons consuming olestra potato chips. AB - BACKGROUND: Olestra is a nonabsorbable fat substitute that consists of fatty acids esterified to a sucrose molecule. OBJECTIVE: To determine the effect of olestra consumption on measurements of fecal fat excretion. DESIGN: Controlled cross-over trial. SETTING: Clinical research center and outpatient research laboratory. PARTICIPANTS: 10 healthy volunteers. INTERVENTION: On days 1 to 6 of the study, participants consumed 5 oz of conventional potato chips per day; on days 7 to 12, they consumed 5 oz of potato chips containing 40 g of olestra per day. MEASUREMENTS: Quantitative measurement of fecal fat by the van de Kamer titration, van de Kamer gravimetric, and Jeejeebhoy gravimetric methods and qualitative assessment of fecal fat by Sudan III staining. RESULTS: Excellent correlation was seen among the three quantitative assays, but the van de Kamer titration method yielded lower measurements than the two gravimetric methods. When participants consumed 40 g of olestra per day, the excretion of fecal fat increased to levels observed in patients with steatorrhea caused by the malabsorption syndrome. CONCLUSION: Consumption of olestra can cause false positive results on tests for steatorrhea and may therefore lead to an erroneous diagnosis of the malabsorption syndrome. PMID- 10681283 TI - Fatal hyperammonemia after orthotopic lung transplantation. AB - BACKGROUND: A case of fatal hyperammonemia complicating orthotopic lung transplantation was previously reported. OBJECTIVE: To describe the incidence, clinical features, and treatment of hyperammonemia associated with orthotopic lung transplantation. DESIGN: Retrospective cohort analysis. SETTING: Academic medical center and lung transplantation center in Philadelphia, Pennsylvania. PATIENTS: 145 sequential adult patients who underwent orthotopic lung transplantation. MEASUREMENTS: Plasma ammonium levels. RESULTS: Six of the 145 patients who had had orthotopic lung transplantation developed hyperammonemia, all within the first 26 days after transplantation. The 30-day post transplantation mortality rate was 67% for patients with hyperammonemia compared with 17% for those without hyperammonemia (P = 0.01). Development of major gastrointestinal complications (P = 0.03), use of total parenteral nutrition (P < 0.001), and lung transplantation for primary pulmonary hypertension (P = 0.045) were associated with hyperammonemia. CONCLUSIONS: Hyperammonemia is a potentially fatal event occurring after orthotopic lung transplantation. It is associated with high nitrogen load, concurrent medical stressors, primary pulmonary hypertension, and hepatic glutamine synthetase deficiency. PMID- 10681284 TI - Update in critical care medicine. PMID- 10681285 TI - Pathogenesis, natural history, treatment, and prevention of hepatitis C. AB - Approximately 4 million persons in the United States and probably more than 100 million persons worldwide are infected with hepatitis C virus. The virus has the unique ability to cause persistent infection in susceptible hosts after parenteral or percutaneous transmission, and its underlying mechanisms are not well understood. The immunologic correlates of protection and viral clearance and the pathogenesis of liver injury are yet to be defined, but recent studies suggest the importance of cell-mediated immune responses. Although 70% to 80% of infected persons become chronic carriers, most have relatively mild disease with slow progression. However, chronic and progressive hepatitis C carries significant morbidity and mortality and is a major cause of cirrhosis, end-stage liver disease, and liver cancer. Development of an effective hepatitis C virus vaccine is not imminent, but recent advances in technology and basic knowledge of molecular virology and immunology have engendered novel approaches to the fundamental problems encountered in vaccine development. Current therapy for hepatitis C, although effective in some patients, is problematic and still evolving. Advances in modern biology and immunology promise new therapies for this important disease. PMID- 10681286 TI - The case for more cautious, patient-focused antiretroviral therapy. AB - Many clinicians who care for patients with HIV infection are dissatisfied with the existing recommendations on antiretroviral therapy. Current practice focuses on the early suppression of viremia, yet the outcome of that approach may not be in the best interest of individual patients or populations. The major goal of HIV therapy is to maintain the long-term health of the patient while avoiding drug related toxicity and preserving viable future treatment options. Recent studies have challenged the principles on which recommendations for early, aggressive treatment were based. Key studies that lead to licensure of antiretroviral medications usually involve short-term results in treatment-naive patients; it is difficult to apply these results to long-term management of therapy-experienced patients. Early, aggressive therapy often prematurely exposes patients to risks for medication-related side effects and resistance. A more cautious, patient focused, long-term approach to therapy would help foster studies of alternate strategies, such as delayed initiation of therapy, protease-sparing therapy, class-sparing therapy, planned drug interruptions, switches in therapy, and immune-based therapy. It is time for clinicians to rethink their approach to the treatment of HIV infection. PMID- 10681287 TI - Eugenic sterilization and a qualified Nazi analogy: the United States and Germany, 1930-1945. AB - In the United States and Germany before World War II, physicians participated in state-authorized eugenic sterilization programs in an attempt to prevent persons deemed to possess undesirable heritable characteristics from propagating. A comparison of U.S. and German histories reveals similarities that argue against easy dismissal of a Nazi analogy. On the basis of a review of editorials in New England Journal of Medicine and Journal of the American Medical Association from 1930 to 1945 it is difficult to accept the suggestion that the alliance between the medical profession and the eugenics movement in the United States was short lived. Comparison of the histories of the eugenic sterilization campaigns in the United States and Nazi Germany reveals important similarities of motivation, intent, and strategy and differences that explain why support for eugenic sterilization in the United States gradually weakened. The eugenics movement in Germany was influenced by economic crisis, radical nationalism, Hitler's totalitarianism, and the medical profession's willing participation and attraction to Nazism for financial and ideological reasons. In the United States, a combination of public unease, Roman Catholic opposition, federal democracy, judicial review, and critical scrutiny by the medical profession reversed the momentum of the eugenics movement and led to the conclusion that eugenic sterilization should be voluntary. PMID- 10681288 TI - Antiretroviral therapy: time to think strategically. PMID- 10681289 TI - The eyes of the Asp. PMID- 10681290 TI - Functional somatic syndromes. PMID- 10681291 TI - Functional somatic syndromes. PMID- 10681292 TI - Functional somatic syndromes. PMID- 10681293 TI - Functional somatic syndromes. PMID- 10681294 TI - Functional somatic syndromes. PMID- 10681296 TI - Functional somatic syndromes. PMID- 10681295 TI - Functional somatic syndromes. PMID- 10681297 TI - Functional somatic syndromes. PMID- 10681298 TI - Functional somatic syndromes. PMID- 10681299 TI - Protease inhibitors do not interfere with prohormone processing. PMID- 10681300 TI - Rapid identification of pathogens in blood. PMID- 10681301 TI - The role of integrin-mediated cell adhesion in health and disease: integrin-based therapy in clinical medicine. PMID- 10681302 TI - In celebration of a life: a meeting of his fellows and friends. John J. Conley memorial. New York City, New York, USA. November 19, 1999. PMID- 10681303 TI - Reduced inhibitory capacity in prefrontal cortex of schizophrenics. PMID- 10681304 TI - Neural circuitry of the prefrontal cortex in schizophrenia. PMID- 10681305 TI - In pursuit of the molecular neuropathology of schizophrenia. PMID- 10681306 TI - Carrier-mediated delivery of 9-(2-phosphonylmethoxyethyl)adenine to parenchymal liver cells: a novel therapeutic approach for hepatitis B. AB - Our aim is to selectively deliver 9-(2-phosphonylmethoxyethyl)adenine (PMEA) to parenchymal liver cells, the primary site of hepatitis B virus (HBV) infection. Selective delivery is necessary because PMEA, which is effective against HBV in vitro, is hardly taken up by the liver in vivo. Lactosylated reconstituted high density lipoprotein (LacNeoHDL), a lipid particle that is specifically internalized by parenchymal liver cells via the asialoglycoprotein receptor, was used as the carrier. PMEA could be incorporated into the lipid moiety of LacNeoHDL by attaching, via an acid-labile bond, lithocholic acid-3alpha-oleate to the drug. The uptake of the lipophilic prodrug (PMEA-LO) by the liver was substantially increased after incorporation into LacNeoHDL. Thirty minutes after injection of [(3)H]PMEA-LO-loaded LacNeoHDL into rats, the liver contained 68.9% +/- 7.7% of the dose (free [(3)H]PMEA, <5%). Concomitantly, the uptake by the kidney was reduced to <2% of the dose (free [(3)H]PMEA, >45%). The hepatic uptake of PMEA-LO-loaded LacNeoHDL occurred mainly by parenchymal cells (88.5% +/- 8.2% of the hepatic uptake). Moreover, asialofetuin inhibited the liver association by >75%, indicating uptake via the asialoglycoprotein receptor. The acid-labile linkage in PMEA-LO, designed to release PMEA during lysosomal processing of the prodrug-loaded carrier, was stable at physiological pH but was hydrolyzed at lysosomal pH (half-life, 60 to 70 min). Finally, subcellular fractionation indicates that the released PMEA is translocated to the cytosol, where it is converted into its active diphosphorylated metabolite. In conclusion, lipophilic modification and incorporation of PMEA into LacNeoHDL improves the biological fate of the drug and may lead to an enhanced therapeutic efficacy against chronic hepatitis B. PMID- 10681307 TI - Nasal carriage in Vietnamese children of Streptococcus pneumoniae resistant to multiple antimicrobial agents. AB - Resistance to antimicrobial agents in Streptococcus pneumoniae is increasing rapidly in many Asian countries. There is little recent information concerning resistance levels in Vietnam. A prospective study of pneumococcal carriage in 911 urban and rural Vietnamese children, of whom 44% were nasal carriers, was performed. Carriage was more common in children <5 years old than in those >/=5 years old (192 of 389 [49.4%] versus 212 of 522 [40.6%]; P, 0.01). A total of 136 of 399 isolates (34%) had intermediate susceptibility to penicillin (MIC, 0.1 to 1 mg/liter), and 76 of 399 isolates (19%) showed resistance (MIC, >1.0 mg/liter). A total of 54 of 399 isolates (13%) had intermediate susceptibility to ceftriaxone, and 3 of 399 isolates (1%) were resistant. Penicillin resistance was 21.7 (95% confidence interval, 7.0 to 67.6) times more common in urban than in rural children (35 versus 2%; P, <0.001). More than 40% of isolates from urban children were also resistant to erythromycin, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline. Penicillin resistance was independently associated with an urban location when the age of the child was controlled for. Multidrug resistance (resistance to three or more antimicrobial agent groups) was present in 32% of isolates overall but in 39% of isolates with intermediate susceptibility to penicillin and 86% of isolates with penicillin resistance. The predominant serotypes of the S. pneumoniae isolates were 19, 23, 14, 6, and 18. Almost half of the penicillin-resistant isolates serotyped were serotype 23, and these isolates were often multidrug resistant. This study suggests that resistance to penicillin and other antimicrobial agents is common in carriage isolates of S. pneumoniae from children in Vietnam. PMID- 10681308 TI - In vitro activities of novel trans-3,5-disubstituted pyrrolidinylthio-1beta methylcarbapenems with potent activities against methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa. AB - The in vitro activities of the novel 1beta-methylcarbapenems J-111, 225, J 114,870, and J-114,871, which have a structurally unique side chain that consists of a trans-3,5-disubstituted 5-arylpyrrolidin-3-ylthio moiety at the C-2 position, were compared with those of reference antibiotics. Among isolates of both methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS), 90% were inhibited by J-111,347 (prototype), J-111,225, J-114,870, and J-114,871 at concentrations of 2, 4, 4, and 4 microgram/ml (MICs at which 90% of isolates are inhibited [MIC(90)s]), respectively, indicating that these agents were 32- to 64-fold more potent than imipenem, which has an MIC(90) of 128 microgram/ml. Although these drugs were less active in vitro than vancomycin, which had MIC(90)s of 1 and 2 microgram/ml for MRSA and MRCoNS, respectively, the new carbapenems displayed better killing kinetics than vancomycin. The potent anti-MRSA activity was ascribed to the excellent affinities of the new carbapenems for penicillin-binding protein 2a of MRSA. Since the new carbapenems also exhibited good activity against gram positive and -negative bacteria including clinically important pathogens such as penicillin-resistant Streptococcus pneumoniae, Haemophilus influenzae, members of the family Enterobacteriaceae, Pseudomonas aeruginosa, and Clostridium difficile, as well as MRSA, the novel carbapenems are worthy of further evaluation. PMID- 10681309 TI - Differential effects of antiretroviral nucleoside analogs on mitochondrial function in HepG2 cells. AB - Numerous studies have reported effects of antiviral nucleoside analogs on mitochondrial function, but they have not correlated well with the observed toxic side effects. By comparing the effects of the five Food and Drug Administration approved anti-human immunodeficiency virus nucleoside analogs, zidovudine (3' azido-3'-deoxythymidine) (AZT), 2',3'-dideoxycytidine (ddC), 2', 3' dideoxyinosine (ddI), 2',3'-didehydro-2',3'-deoxythymidine (d4T), and beta-L 2',3'-dideoxy-3'-thiacytidine (3TC), as well as the metabolite of AZT, 3'-amino 3'-deoxythymidine (AMT), on mitochondrial function in a human hepatoma cell line, this issue has been reexamined. Evidence for a number of mitochondrial defects with AZT, ddC, and ddI was found, but only AZT induced a marked rise in lactic acid levels. Only in mitochondria isolated from AZT (50 microM)-treated cells was significant inhibition of cytochrome c oxidase and citrate synthase found. Our investigations also demonstrated that AZT, d4T, and 3TC did not affect the synthesis of the 11 polypeptides encoded by mitochondrial DNA, while ddC caused 70% reduction of total polypeptide content and ddI reduced by 43% the total content of 8 polypeptides (including NADH dehydrogenase subunits 1, 2, 4, and 5, cytochrome c oxidase subunits I to III, and cytochrome b). We hypothesize that in hepatocytes the reserve capacity for mitochondrial respiration is such that inhibition of respiratory enzymes is unlikely to become critical. In contrast, the combined inhibition of the citric acid cycle and electron transport greatly enhances the dependence of the cell on glycolysis and may explain why apparent mitochondrial dysfunction is more prevalent with AZT treatment. PMID- 10681310 TI - Consequences of interaction of a lipophilic endotoxin antagonist with plasma lipoproteins. AB - E5531, a novel synthetic lipid A analogue, antagonizes the toxic effects of lipopolysaccharide, making it a potential intravenously administered therapeutic agent for the treatment of sepsis. This report describes the distribution of E5531 in human blood and its activity when it is associated with different lipoprotein subclasses. After in vitro incubation of [(14)C]E5531 with blood, the great majority (>92%) of material was found in the plasma fraction. Analysis by size-exclusion and affinity chromatographies and density gradient centrifugation indicates that [(14)C]E5531 binds to lipoproteins, primarily high-density lipoproteins (HDLs), with distribution into low-density lipoproteins (LDLs) and very low density lipoproteins (VLDLs) being dependent on the plasma LDL or VLDL cholesterol concentration. Similar results were also seen in a limited study of [(14)C]E5531 administration to human volunteers. The potency of E5531 in freshly drawn human blood directly correlates to increasing LDL cholesterol levels. Finally, preincubation of E5531 with plasma or purified lipoproteins indicated that binding to HDL resulted in a time-dependent loss of drug activity. This loss in activity was not observed with drug binding to LDLs or to VLDLs or chylomicrons. Taken together, these results indicate that E5531 binds to plasma lipoproteins, making its long-term antagonistic potency dependent on the plasma lipoprotein composition. PMID- 10681311 TI - Effect of proinflammatory cytokines on the interplay between roxithromycin, HMR 3647, or HMR 3004 and human polymorphonuclear neutrophils. AB - Cytokines, the hallmarks of infectious and inflammatory diseases, modify phagocyte activities and thus may interfere with the immunomodulating properties of antibacterial agents. We have investigated whether various proinflammatory cytokines (interleukin 1 [IL-1], IL-6, IL-8, gamma interferon, tumor necrosis factor alpha [TNF-alpha], and granulocyte-macrophage colony-stimulating factor [GM-CSF]) modify two macrolide properties, i.e., inhibition of oxidant production by polymorphonuclear neutrophils (PMN) and cellular uptake. Roxithromycin and two ketolides, HMR 3647 and HMR 3004, were chosen as the test agents. TNF-alpha and GM-CSF (but not the other cytokines) decreased the inhibitory effect of HMR 3647 only on oxidant production by PMN. Fifty percent inhibitory concentrations were, however, in the same range in control and cytokine-treated cells (about 60 to 70 microgram/ml), suggesting that HMR 3647 acts downstream of the priming effect of cytokines. In contrast, the impairment of oxidant production by roxithromycin and HMR 3004 was unchanged (or increased) in cytokine-treated cells. This result suggests that HMR 3004 (the strongest inhibitory drug, likely owing to its quinoline side chain) and roxithromycin act on a cellular target upstream of cytokine action. In addition, TNF-alpha and GM-CSF significantly (albeit moderately) impaired (by about 20%) the uptake of the three molecules by PMN. The inhibitory effect of these two cytokines seems to be related to activation of the p38 mitogen-activated protein kinase. Our data also illuminate the mechanism underlying macrolide uptake: protein kinase A- and tyrosine kinase-dependent phosphorylation seems to be necessary for optimal uptake, while protein kinase C activation impairs it. The relevance of our data to the clinical setting requires further investigations, owing to the complexity of the cytokine cascade during infection and inflammation. PMID- 10681312 TI - Mechanism of uptake of a cationic water-soluble pyridinium zinc phthalocyanine across the outer membrane of Escherichia coli. AB - Previous studies have shown that a cationic water-soluble pyridinium zinc phthalocyanine (PPC) is a powerful photosensitizer that is able to inactivate Escherichia coli. In the current work incubation of E. coli cells with PPC in the dark caused alterations in the outer membrane permeability barrier of the cells, rendering the bacteria much more sensitive to hydrophobic compounds, with little effect seen with hydrophilic compounds. Addition of Mg(2+) to the medium prior to incubation of the cells with PPC prevented these alterations in the outer membrane permeability barrier. The presence of Mg(2+) in the medium also prevented the photoinactivation of E. coli cells with PPC. These results are consistent with the hypothesis that PPC gains access across the outer membrane of E. coli cells via the self-promoted uptake pathway, a mechanism of uptake postulated for the uptake of other cationic compounds across the outer membranes of gram-negative bacteria. PMID- 10681313 TI - pncA mutations as a major mechanism of pyrazinamide resistance in Mycobacterium tuberculosis: spread of a monoresistant strain in Quebec, Canada. AB - Pyrazinamide (PZA) is an important first-line tuberculosis drug that is part of the currently used short-course tuberculosis chemotherapy. PZA is a prodrug that has to be converted to the active form pyrazinoic acid by pyrazinamidase (PZase) activity, encoded by the pncA gene of Mycobacterium tuberculosis, and loss of PZase activity is associated with PZA resistance. To further define the genetic basis of PZA resistance and determine the frequency of PZA-resistant strains having pncA mutations, we sequenced the pncA gene from a panel of 59 PZA resistant clinical isolates from Canada, the United States, and Korea. Two strains that did not contain pncA mutations and had positive PZase turned out to be falsely resistant. Three PZase-negative strains (MIC, >900 microgram of PZA per ml) and one PZase-positive strain (strain 9739) (MIC, >300 microgram of PZA per ml) did not have pncA mutations. The remaining 53 of the 57 PZA-resistant isolates had pncA mutations, confirming that pncA mutation is the major mechanism of PZA resistance. Various new and diverse mutations were found in the pncA gene. Interestingly, 20 PZA-monoresistant strains and 1 multidrug-resistant isolate from Quebec, Canada, all had the same pncA mutation profile, consisting of an 8 nucleotide deletion and an amino acid substitution of Arg140-->Ser. Strain typing indicated that these strains are highly related and share almost identical IS6110 patterns. These data strongly suggest the spread of a PZA-monoresistant strain, which has not previously been described. PMID- 10681314 TI - Antibacterial efficacy of gentamicin encapsulated in pH-sensitive liposomes against an in vivo Salmonella enterica serovar typhimurium intracellular infection model. AB - Encapsulation of gentamicin in liposomes can be used to achieve intracellular delivery and broaden the clinical utility of this drug. We have previously described a novel, rationally designed, pH-sensitive liposomal carrier for gentamicin that has superior in vitro efficacy against intracellular infections compared to the efficacies of both free gentamicin and non-pH-sensitive liposomal controls. This liposomal carrier demonstrated pH-sensitive fusion that was dependent on the presence of unsaturated phosphatidylethanolamine (PE) and the pH sensitive lipid N-succinyldioleoyl-PE. The pharmacokinetics and biodistribution of the free and liposomal gentamicin were examined in mice bearing a systemic Salmonella enterica serovar Typhimurium infection. Encapsulation of gentamicin in pH-sensitive liposomes significantly increased the concentrations of the drug in plasma compared to those of free gentamicin. Furthermore, the levels of accumulation of drug in the infected liver and spleen were increased by 153- and 437-fold, respectively, as a result of liposomal encapsulation. The increased accumulation of gentamicin in the liver and spleen effected by liposomal delivery was associated with 10(4)-fold greater antibacterial activity than that associated with free gentamicin in a murine salmonellosis model. These pH sensitive liposomal antibiotic carriers with enhanced in vitro activity could be used to improve both in vivo intracellular drug delivery and biological activity. PMID- 10681315 TI - Efficacy of liposomal amphotericin B with prolonged circulation in blood in treatment of severe pulmonary aspergillosis in leukopenic rats. AB - The therapeutic efficacy of long-circulating polyethylene glycol-coated liposomal amphotericin B (AMB) (PEG-AMB-LIP) was compared with that of AMB desoxycholate (Fungizone) in a model of severe invasive pulmonary aspergillosis in persistently leukopenic rats as well as in temporarily leukopenic rats. PEG-AMB-LIP treatment (intravenous administration) consisted of a single, or double (every 72 h), or triple (every 72 h) dose of 10 mg of AMB/kg of body weight, a double dose (every 72 h) of 14 mg of AMB/kg, or a 5-day treatment (every 24 h) with 6 mg/kg/dose. AMB desoxycholate was administered for 10 consecutive days at 1 mg of AMB/kg/dose. Treatment was started 30 h after fungal inoculation, at which time mycelial growth was firmly established. Both persistently and temporarily leukopenic rats died between 4 and 9 days after Aspergillus fumigatus inoculation when they were left untreated or after treatment with a placebo. In persistently leukopenic rats, a single dose of PEG-AMB-LIP (10 mg/kg) was as effective as the 10-day treatment with AMB desoxycholate (at 1 mg/kg/dose) in significantly prolonging the survival of rats infected with A. fumigatus and in reducing the dissemination of A. fumigatus to the liver. Prolongation of PEG-AMB-LIP treatment (double or triple dose or 5-day treatment) did not further improve efficacy. For temporarily leukopenic rats no major advances in efficacy were achieved compared to those for persistently leukopenic rats, probably because the leukocyte numbers in blood were restored too late in the course of infection. PMID- 10681316 TI - A sensitive amphotericin B immunoassay for pharmacokinetic and distribution studies. AB - Since currently used assays of amphotericin B (AMB) lack sensitivity or are not easily adaptable in all laboratories, we have developed an enzyme immunoassay for AMB in biological fluids and tissues. Antibodies to AMB were raised in rabbits after administration of an AMB-bovine serum albumin conjugate. The enzymatic tracer was obtained by coupling AMB via its amino group to acetylcholinesterase (EC 3.1.1.7). These reagents were used for the development of a competitive immunoassay performed on microtitration plates. The limit of quantification was 100 pg/ml in plasma and 1 ng/g in tissues. The plasma assay was performed directly without extraction on a minimal volume of 0.1 ml. The intra- and interassay coefficients of variation were in the range of 5 to 17%, and the recoveries were 92 to 111% for AMB added to human plasma. The assay was applied to a pharmacokinetic study with mice given AMB intraperitoneally at the dose of 1 mg/kg. The drug distribution in selected compartments (plasma, liver, spleen, lung, and brain) was monitored until 72 h after administration. In conclusion, our assay is at least 100-fold more sensitive than previously described bioassays or chromatographic determinations of AMB and may be useful in studying the tissue pharmacokinetics of new AMB formulations and in drug monitoring in clinical situations. PMID- 10681317 TI - In vitro antihepadnaviral activities of combinations of penciclovir, lamivudine, and adefovir. AB - Penciclovir (9-[2-hydroxy-1-(hydroxymethyl)-ethoxymethyl]guanine [PCV]), lamivudine ([-]-beta-L-2',3'-dideoxy-3'-thiacytidine [3TC]), and adefovir (9-[2 phosphonylmethoxyethyl]-adenine [PMEA]) are potent inhibitors of hepatitis B virus (HBV) replication. Lamivudine has recently received approval for clinical use against chronic human HBV infection, and both PCV and PMEA have undergone clinical trials against HBV in their respective prodrug forms (famciclovir and adefovir dipivoxil [bis-(POM)-PMEA]). Since multidrug combinations are likely to be used to control HBV infection, investigation of potential interactions between PCV, 3TC, and PMEA is important. Primary duck hepatocyte cultures which were either acutely or congenitally infected with the duck hepatitis B virus (DHBV) were used to investigate in vitro interactions between PCV, 3TC, and PMEA. Here we show that the anti-DHBV effects of all the combinations containing PCV, 3TC, and PMEA are greater than that of each of the individual components and that their combined activities are approximately additive or synergistic. These results may underestimate the potential in vivo usefulness of PMEA-containing combinations, since there is evidence that PMEA has immunomodulatory activity and, at least in the duck model of chronic HBV infection, is capable of inhibiting DHBV replication in cells other than hepatocytes, the latter being unaffected by treatment with either PCV or 3TC. Further investigation of the antiviral activities of these drug combinations is therefore required, particularly since each of the component drugs is already in clinical use. PMID- 10681318 TI - ampR gene mutations that greatly increase class C beta-lactamase activity in Enterobacter cloacae. AB - The ampC and ampR genes of Enterobacter cloacae GN7471 were cloned into pMW218 to yield pKU403. Four mutant plasmids derived from pKU403 (pKU404, pKU405, pKU406, and pKU407) were isolated in an AmpD mutant of Escherichia coli ML4953 by selection with ceftazidime or aztreonam. The beta-lactamase activities expressed by pKU404, pKU405, pKU406, and pKU407 were about 450, 75, 160, and 160 times higher, respectively, than that expressed by the original plasmid, pKU403. These mutant plasmids all carried point mutations in the ampR gene. In pKU404 and pKU405, Asp-135 was changed to Asn and Val, respectively. In both pKU406 and pKU407, Arg-86 was changed to Cys. The ease of selection of AmpR mutations at a frequency of about 10(-6) in this study strongly suggests that derepressed strains, such as AmpD or AmpR mutants, could frequently emerge in the clinical setting. PMID- 10681319 TI - A novel human immunodeficiency virus type 1 reverse transcriptase mutational pattern confers phenotypic lamivudine resistance in the absence of mutation 184V. AB - We describe a new human immunodeficiency virus type 1 (HIV-1) mutational pattern associated with phenotypic resistance to lamivudine (3TC) in the absence of the characteristic replacement of methionine by valine at position 184 (M184V) of reverse transcriptase. Combined genotypic and phenotypic analyses of clinical isolates revealed the presence of moderate levels of phenotypic resistance (between 4- and 50-fold) to 3TC in a subset of isolates that did not harbor the M184V mutation. Mutational cluster analysis and comparison with the phenotypic data revealed a significant correlation between moderate phenotypic 3TC resistance and an increased incidence of replacement of glutamic acid by aspartic acid or alanine and of valine by isoleucine at residues 44 and 118 of reverse transcriptase, respectively. This occurred predominantly in those isolates harboring zidovudine resistance-associated mutations (41L, 215Y). The requirement of the combination of mutations 41L and 215Y with mutations 44D and 44A and/or 118I for phenotypic 3TC resistance was confirmed by site-directed mutagenesis experiments. These data support the assumption that HIV-1 may have access to several different genetic pathways to escape drug pressure or that the increase in the frequency of particular mutations may affect susceptibility to drugs that have never been part of a particular regimen. PMID- 10681320 TI - Faropenem transport across the renal epithelial luminal membrane via inorganic phosphate transporter Npt1. AB - We previously showed that the mouse inorganic phosphate transporter Npt1 operates in the hepatic sinusoidal membrane transport of anionic drugs such as benzylpenicillin and mevalonic acid. In the present study, the mechanism of renal secretion of penem antibiotics was examined by using a Xenopus oocyte expression system. Faropenem (an oral penem antibiotic) was transported via Npt1 with a Michaelis-Menten constant of 0.77 +/- 0.34 mM in a sodium-independent but chloride ion-sensitive manner. When the concentration of chloride ions was increased, the transport activity of faropenem by Npt1 was decreased. Since the concentration gradient of chloride ions is in the lumen-to-intracellular direction, faropenem is expected to be transported from inside proximal tubular cells to the lumen. So, we tested the release of faropenem from Xenopus oocytes. The rate of efflux of faropenem from Npt1-expressing oocytes was about 9.5 times faster than that from control water-injected Xenopus oocytes. Faropenem transport by Npt1 was significantly inhibited by beta-lactam antibiotics such as benzylpenicillin, ampicillin, cephalexin, and cefazolin to 24.9, 40. 5, 54.4, and 26.2% of that for the control, respectively. Zwitterionic beta-lactam antibiotics showed lesser inhibitory effects on faropenem uptake than anionic derivatives, indicating that Npt1 preferentially transports anionic compounds. Other anionic compounds, such as indomethacin and furosemide, and the anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid significantly inhibited faropenem uptake mediated by Npt1. In conclusion, our results suggest that Npt1 participates in the renal secretion of penem antibiotics. PMID- 10681321 TI - Pharmacokinetics and safety of ascending single doses of DZ-2640, a new oral carbapenem antibiotic, administered to healthy Japanese subjects. AB - DZ-2640 is the ester-type oral carbapenem prodrug of an active parent compound, DU-6681. The pharmacokinetics and safety of DU-6681 were investigated in six studies after oral administration of a single dose of DZ-2640 to healthy male Japanese volunteers at doses of 25, 50, 100, 200, and 400 mg (as the equivalents of DU-6681) in the fasted state. The same volunteers received the drug at a dose of 100 mg in the fasted and fed states to examine the effect of food intake on the bioavailability of DZ-2640. The concentrations of DU-6681 in plasma and urine were determined by a validated high-performance liquid chromatography method and a bioassay. A good correlation between both methods was seen, indicating an absence of major active metabolites. The mean maximum concentrations of DU-6681 in plasma (C(max)) ranged from 0.263 microgram/ml (25-mg dose) to 2.489 microgram/ml (400-mg dose) and were reached within 1.5 h following drug administration. After reaching the C(max), plasma DU-6681 concentrations declined in a monophasic manner, with a half-life of 0.47 to 0.89 h. The area under the concentration-time curve (AUC) and C(max) increased almost linearly with the dose up to the 200-mg dose. The AUC and C(max) increased less than proportionally after administration of the 400-mg dose, suggesting a reduction in drug absorption. The plasma protein binding of DU-6681 was in the range of 23.3 to 25.6%. The cumulative urinary recoveries (0 to 24 h) were in the range of 31.9 to 44.9%. The AUC was slightly but statistically significantly reduced by food intake. However, the C(max), half-life, and recovery in urine were not affected by food intake. The renal clearance (402 to 510 ml/min) was much greater than the mean glomerular filtration rate (ca. 120 ml/min), which indicated active tubular secretion of the drug. A mild transient and moderate diarrhea was observed in two of six volunteers in the study with a single dose of 25 mg. Mild soft stools were observed in two of six volunteers who received a 400-mg dose of the drug. PMID- 10681322 TI - Inactivation of the ampD gene in Pseudomonas aeruginosa leads to moderate-basal level and hyperinducible AmpC beta-lactamase expression. AB - It has been shown in enterobacteria that mutations in ampD provoke hyperproduction of chromosomal beta-lactamase, which confers to these organisms high levels of resistance to beta-lactam antibiotics. In this study, we investigated whether this genetic locus was implicated in the altered AmpC beta lactamase expression of selected clinical isolates and laboratory mutants of Pseudomonas aeruginosa. The sequences of the ampD genes and promoter regions from these strains were determined and compared to that of wild-type ampD from P. aeruginosa PAO1. Although we identified numerous nucleotide substitutions, they resulted in few amino acid changes. The phenotypes produced by these mutations were ascertained by complementation analysis. The data revealed that the ampD genes of the P. aeruginosa mutants transcomplemented Escherichia coli ampD mutants to the same levels of beta-lactam resistance and beta-lactamase expression as wild-type ampD. Furthermore, complementation of the P. aeruginosa mutants with wild-type ampD did not restore the inducibility of beta-lactamase to wild-type levels. This shows that the amino acid substitutions identified in AmpD do not cause the altered phenotype of AmpC beta-lactamase expression in the P. aeruginosa mutants. The effects of AmpD inactivation in P. aeruginosa PAO1 were further investigated by gene replacement. This resulted in moderate-basal-level and hyperinducible expression of beta-lactamase accompanied by high levels of beta-lactam resistance. This differs from the stably derepressed phenotype reported in AmpD-defective enterobacteria and suggests that further change at another unknown genetic locus may be causing total derepressed AmpC production. This genetic locus could also be altered in the P. aeruginosa mutants studied in this work. PMID- 10681323 TI - A dose ranging study of the pharmacokinetics, safety, and preliminary efficacy of lamivudine in children and adolescents with chronic hepatitis B. AB - Fifty-three patients with chronic hepatitis B and active viral replication were studied for 4 weeks while on treatment and for 12 weeks after treatment with the oral nucleoside analogue lamivudine. Children aged 2 to 12 years were randomized to receive twice-daily doses of 0.35, 1.5, or 4 mg of lamivudine solution per kg of body weight or once-daily doses of 3 mg of lamivudine solution per kg. Adolescents aged 13 to 17 years received lamivudine at 100 mg (as tablets). Blood samples for pharmacokinetic assay were taken on days 1 and 28. Lamivudine was rapidly absorbed following oral administration, with the maximum concentration in serum being reached 0.5 to 1 h postdosing. Apparent oral clearance (CL/F) was higher in younger children and decreased with age, with CL/F values for adolescents reaching those seen for adults by the age of 12. All doses produced a dramatic fall in serum hepatitis B virus (HBV) DNA levels, with a median reduction of >/=99.5% after 4 weeks of treatment and with the levels returning to the baseline levels posttreatment. The correlation of dose, area under the concentration-time curve (AUC), and changes in HBV DNA levels, as measured by the Chiron Quantiplex assay, showed maximal antiviral effects (99.9% inhibition and a reduction of the amount of HBV DNA of approximately 3 log(10)) at 3 mg/kg/day, with no discernible increase in effect seen whether the drug was given at 4 mg/kg twice daily or whether it was given once daily or twice daily. The limit of detection of the assay (2.5 pg/ml) was reached for some but not all patients across the dose ranges, with the smallest number (n = 2) of those having values negative by the Chiron Quantiplex assay being in the lowest-dose group. The 13- to 17-year-olds showed a similar overall response in terms of the HBV DNA level reduction compared to that for patients younger than age 13. Analysis of the same samples by PCR, which has a lower limit of sensitivity than the Chiron Quantiplex assay, also showed average drops in HBV DNA levels of about 3 log(10) at 4 weeks for patients for which the AUC was >/=4,000 ng. h/ml, confirming the conclusions given above. Lamivudine treatment was well tolerated at all doses, with no significant adverse events or laboratory data changes. On the basis of pharmacokinetic and pharmacodynamic data, a 3-mg/kg/day dose in children (ages 2 to 12 years) with chronic hepatitis B provides levels of exposure and trough concentrations similar to those seen in adults following the receipt of doses of 100 mg. The 100-mg dose is being evaluated in a large phase III study with HBV infected pediatric patients. PMID- 10681324 TI - Activities of trovafloxacin, gatifloxacin, clinafloxacin, sparfloxacin, levofloxacin, and ciprofloxacin against penicillin-resistant Streptococcus pneumoniae in an in vitro infection model. AB - We adapted an in vitro pharmacodynamic model of infection to incorporate infected fibrin clots. The bactericidal activities of various fluoroquinolones against two strains of penicillin-resistant Streptococcus pneumoniae were studied over a 48-h period. Bacteria were prepared in Muller-Hinton broth by using colonies from a 24 h tryptic soy agar plus 5% sheep blood plate and were added to a mixture of cryoprecipitate (80%) and thrombin (10%) to achieve approximately 10(6) CFU of organism per fibrin clot. The fibrin clots were suspended into the models and removed, in triplicate, at various time points over 48 h. Control models were also conducted to characterize the growth of S. pneumoniae in the growth medium without antibiotic. Trovafloxacin, gatifloxacin, clinafloxacin, sparfloxacin, levofloxacin, and ciprofloxacin were administered to simulate their pharmacokinetic profiles in humans. Fibrin clot samples were also plated onto antibiotic-containing tryptic soy agar plus 5% lysed horse blood to detect resistance. The newer fluoroquinolones demonstrated better activity than ciprofloxacin against both isolates. In conclusion, the newer quinolones demonstrated significant activity against penicillin-resistant S. pneumoniae, with standard dosing resulting in area under the concentration-time curve/MIC ratios and peak concentration/MIC ratios that resulted in 99.9% killing against these isolates. PMID- 10681325 TI - Antibacterial and antimembrane activities of cecropin A in Escherichia coli. AB - The ability of cecropin A to permeabilize and depolarize the membranes of Escherichia coli ML-35p bacteria has been compared to its bactericidal activity in an extension of earlier studies performed on synthetic lipid vesicle membranes (L. Silvestro, K. Gupta, J. H. Weiser, and P. H. Axelsen, Biochemistry 36:11452 11460, 1997). Our results indicate that differences in the concentration dependences of membrane permeabilization and depolarization seen in synthetic vesicles are not manifested in whole bacteria. The concentration dependences of both phenomena roughly correlate with bactericidal activity, suggesting that the bactericidal mechanism of cecropin A is related to membrane permeabilization. PMID- 10681326 TI - Frequent association between alteration of the rdxA gene and metronidazole resistance in French and North African isolates of Helicobacter pylori. AB - Mutations in the rdxA gene have been associated with the acquisition of resistance to metronidazole in Helicobacter pylori. This gene encodes an NADPH nitroreductase whose expression is necessary for intracellular activation of the drug. We wished to examine whether mutations in rdxA were present in resistant H. pylori isolates infecting either French or North African patients. We determined the complete nucleotide sequences of the rdxA genes from seven French and six North African patients infected with paired resistant and sensitive strains. Genotyping by random amplified polymorphic DNA analysis confirmed the close genetic relatedness of the susceptible and resistant isolates from individual biopsies. Eight French and five North African individual resistant strains were also studied. For the French strains, an alteration in rdxA most probably implicated in resistance was found in 10 cases (seven frameshift mutations, two missense mutations, and one deletion of 211 bp). One to three putative missense mutations were identified in four cases, and a missense mutation possibly not implicated in resistance was discovered in the last case. For the North African strains, an alteration in rdxA was found in eight cases (three frameshift mutations, three missense mutations, one deletion of 6 bp, and one insertion of a variant of IS605). Two strains contained putative missense mutations, and no change was observed in rdxA of the last strain. Thus, inactivation of the rdxA gene is frequently, but not always, associated with resistance to metronidazole in French and North African clinical isolates of H. pylori. In addition, a variety of alterations of rdxA are associated with the resistant phenotype. PMID- 10681327 TI - Efficacy of FK463, a new lipopeptide antifungal agent, in mouse models of disseminated candidiasis and aspergillosis. AB - The efficacy of intravenous injection of FK463, a novel water-soluble lipopeptide, was evaluated in mouse models of disseminated candidiasis and aspergillosis and was compared with those of fluconazole (FLCZ) and amphotericin B (AMPH-B). In the candidiasis model, FK463 significantly prolonged the survival of intravenously infected mice at doses of 0.125 mg/kg of body weight or higher. In disseminated candidiasis caused by Candida species, including FLCZ-resistant Candida albicans, FK463 exhibited an efficacy 1.4 to 18 times inferior to that of AMPH-B, with 50% effective doses (ED(50)s) ranging from 0.21 to 1.00 mg/kg and 0.06 to 0.26 mg/kg, respectively, and was much more active than FLCZ. The protective effect of FK463 was not obviously influenced by the fungal inoculum size, the starting time of the treatment, or the immunosuppressed status of the host. The reduction in efficacy was less than that observed with FLCZ or AMPH-B. The efficacy of FK463 was also evaluated in the disseminated candidiasis target organ assay and was compared with those of FLCZ and AMPH-B. Efficacies were evaluated on the basis of a comparison between the mean log(10) CFU in kidneys in the groups treated with antifungal agents and that in control group. A single dose of FK463 at 0.5 mg/kg or higher significantly reduced the viable counts in kidneys compared with the numbers of yeast cells before treatment, and its efficacy was comparable to that of AMPH-B, while FLCZ at 4 mg/kg showed only a suppressive effect on the growth of C. albicans in the kidneys. In the disseminated aspergillosis model, FK463 given at doses of 0.5 mg/kg or higher significantly prolonged the survival of mice infected intravenously with Aspergillus fumigatus conidia. The efficacy of FK463 was about 2 times inferior to that of AMPH-B, with ED(50)s ranging from 0.25 to 0.50 mg/kg and 0.11 to 0.29 mg/kg, respectively. These results indicate that FK463 may be a potent parenterally administered therapeutic agent for disseminated candidiasis and aspergillosis. PMID- 10681328 TI - Efficacy of FK463, a new lipopeptide antifungal agent, in mouse models of pulmonary aspergillosis. AB - The efficacy of FK463, a novel water-soluble lipopeptide, was evaluated in mouse models of pulmonary aspergillosis and was compared with that of amphotericin B (AMPH-B). In the pulmonary aspergillosis models induced by intranasal inoculation, FK463 exhibited good efficacy, with 50% effective doses in the range of 0. 26 to 0.51 mg/kg of body weight; these values were comparable to those of AMPH-B. In an Aspergillus target organ assay with immunosuppressed mice, under conditions of constant plasma levels of FK463, using a subcutaneously implanted osmotic pressure pump, a significant reduction in viable fungal cells was observed at plasma FK463 levels of 0.55 to 0.80 microgram/ml or higher. We conclude that FK463 is highly effective in the treatment of pulmonary aspergillosis in this animal model. These results indicate that FK463 may be a potent parenterally administered antifungal agent for pulmonary aspergillosis. PMID- 10681329 TI - Biochemical sequence analyses of GES-1, a novel class A extended-spectrum beta lactamase, and the class 1 integron In52 from Klebsiella pneumoniae. AB - Klebsiella pneumoniae ORI-1 was isolated in 1998 in France from a rectal swab of a 1-month-old girl who was previously hospitalized in Cayenne Hospital, Cayenne, French Guiana. This strain harbored a ca. 140-kb nontransferable plasmid, pTK1, that conferred an extended-spectrum cephalosporin resistance profile antagonized by the addition of clavulanic acid, tazobactam, or imipenem. The gene for GES-1 (Guiana extended-spectrum beta-lactamase) was cloned, and its protein was expressed in Escherichia coli DH10B, where this pI-5. 8 beta-lactamase of a ca. 31-kDa molecular mass conferred resistance to oxyimino cephalosporins (mostly to ceftazidime). GES-1 is weakly related to the other plasmid-located Ambler class A extended-spectrum beta-lactamases (ESBLs). The highest percentage of amino acid identity was obtained with the carbenicillinase GN79 from Proteus mirabilis; with YENT, a chromosome-borne penicillinase from Yersinia enterocolitica; and with L 2, a chromosome-borne class A cephalosporinase from Stenotrophomonas maltophilia (36% amino acid identity each). However, a dendrogram analysis showed that GES-1 clustered within a class A ESBL subgroup together with ESBLs VEB-1 and PER-1. Sequencing of a 7,098-bp DNA fragment from plasmid pTK1 revealed that the GES-1 gene was located on a novel class 1 integron named In52 that was characterized by (i) a 5' conserved segment containing an intI1 gene possessing two putative promoters, P(1) and P(2), for coordinated expression of the downstream antibiotic resistance genes and an attI1 recombination site; (ii) five antibiotic gene cassettes, bla(GES-1), aac(6')Ib' (gentamicin resistance and amikacin susceptibility), dfrXVb (trimethoprim resistance), a novel chloramphenicol resistance gene (cmlA4), and aadA2 (streptomycin-spectinomycin resistance); and (iii) a 3' conserved segment consisting of qacEDelta1 and sulI. The bla(GES-1) and aadA2 gene cassettes were peculiar, since they lacked a typical 59-base element. This work identified the second class A ESBL gene of a non-TEM, non-SHV series which was located in the plasmid and integron, thus providing it additional means for its spread and its expression. PMID- 10681330 TI - Activities and postantibiotic effects of gemifloxacin compared to those of 11 other agents against Haemophilus influenzae and Moraxella catarrhalis. AB - The activity of gemifloxacin against Haemophilus influenzae and Moraxella catarrhalis was compared to those of 11 other agents. All quinolones were very active (MICs, 50 microgram/ml), had IPC synthase activity (specific activity, 1 to 3 pmol/min/mg of protein) also sensitive to aureobasidin A (IC(50)s, 3 to 5 ng/ml). The mammalian multidrug resistance modulators verapamil, chlorpromazine, and trifluoperazine lowered the MIC of aureobasidin A for A. fumigatus from >50 microgram/ml to 2 to 3 microgram/ml, suggesting that the resistance of this major fungal pathogen is the result of increased efflux. PMID- 10681334 TI - Efficacy of linezolid in experimental otitis media. AB - Therapy for otitis media (OM) due to resistant Streptococcus pneumoniae (MIC of penicillin, >/=2.0 microgram/ml) is challenging. Linezolid, an oxazolidinone, represent a new class of antimicrobial agents with excellent in vitro activity against penicillin-resistant S. pneumoniae; however, in vitro activity against nontypeable Haemophilus influenzae (NTHI) is limited. We evaluated its efficacy against experimental acute OM due to a multidrug-resistant S. pneumoniae isolate and two isolates of NTHI. The chinchilla model was utilized to evaluate the efficacy of linezolid against experimental infection due to S. pneumoniae or NTHI. Serum and middle ear antibiotic concentrations were determined, and sterilization of experimental OM was evaluated. Chinchillas were inoculated directly with S. pneumoniae into the superior bulla. Twenty-four hours after inoculation, all animals had positive middle ear and nasopharyngeal cultures. Animals were given linezolid at 25 mg/kg/dose twice a day (b.i.d.) by orogastric feeding tube or amoxicillin at 40 mg/kg/dose b.i.d. intramuscularly for 5 days. By day 5, all animals in the linezolid group had sterile middle ear cultures and eradication of S. pneumoniae from the nasopharynx. In the amoxicillin group, all nine animals remained middle ear and nasopharynx positive (P < 0.01). In animals inoculated with NTHI, 25 and 37.5 mg/kg b.i.d. failed to sterilize middle ear infection or eradicate colonization. Mean levels in middle ear fluid measured during experimental infection were 12.8 microgram/ml at 2 to 6 h and 4. 1 mirogram/ml at 16 to 17 h after orogastric dosing at 25 mg/kg. Linezolid achieved a high concentration in the middle ear during experimental OM. Linezolid eradicated multidrug-resistant S. pneumoniae from the middle ear and nasopharynx. Experimental infection and nasopharyngeal colonization due to NTHI persisted despite achievement of concentrations in the middle ear that were above the MIC (for NTHI). PMID- 10681335 TI - Assignment of the substrate-selective subunits of the MexEF-OprN multidrug efflux pump of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa expresses a low level of the MexAB-OprM efflux pump and shows natural resistance to many structurally and functionally diverse antibiotics. The mutation that has been referred to previously as nfxC expresses an additional efflux pump, MexEF-OprN, exhibiting resistance to fluoroquinolones, imipenem, and chloramphenicol and hypersusceptibility to beta-lactam antibiotics. To address the antibiotic specificity of the MexEF-OprN efflux pump, we introduced a plasmid carrying the mexEF-oprN operon into P. aeruginosa lacking the mexAB-oprM operon. The transformants exhibited resistance to fluoroquinolones, trimethoprim, and chloramphenicol but, unlike most nfxC-type mutants, did not show beta-lactam hypersusceptibility. The transformants exhibited additional resistance to tetracycline. In the next experiment, we analyzed the MexEF-OprN pump subunit(s) responsible for substrate selectivity by expressing MexE, MexF, OprN, and MexEF in strains lacking MexA, MexB, OprM, and MexAB, respectively. The MexEF-OprM/DeltaMexAB transformants exhibited MexEF-OprN type pump function that rendered the strains resistant to fluoroquinolones and chloramphenicol but did not change susceptibility to beta-lactam antibiotics compared with the host strain. The MexAB-OprN/DeltaOprM, MexAF-OprM/DeltaMexB, and MexEB-OprM/DeltaMexA mutants exhibited antibiotic susceptibility indistinguishable from that in the mutant lacking both types of efflux pumps. The results imply that the MexEF-OprM pump selects substrates by a MexEF functional unit. Interestingly, OprN did not link functionally with the MexAB complex, despite the fact that OprM interacted functionally with MexEF. PMID- 10681336 TI - Apoptosis in renal proximal tubules of rats treated with low doses of aminoglycosides. AB - Kidney cortex apoptosis was studied with female Wistar rats treated for 10 days with gentamicin and netilmicin at daily doses of 10 or 20 mg/kg of body weight and amikacin or isepamicin at daily doses of 40 mg/kg. Apoptosis was detected and quantitated using cytological (methyl green-pyronine) and immunohistochemical (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) staining, in parallel with a measurement of drug-induced phospholipidosis (cortical phospholipids and phospholipiduria), cortical proliferative response ((3)H incorporation in DNA and histoautoradiography after in vivo pulse-labeling with [(3)H]thymidine), and kidney dysfunction (blood urea nitrogen and creatinine). Gentamicin induced in proximal tubules a marked apoptotic reaction which (i) was detectable after 4 days of treatment but was most conspicuous after 10 days, (ii) was dose dependent, (iii) occurred in the absence of necrosis, and (iv) was nonlinearly correlated with the proliferative response (tubular and peritubular cells). Comparative studies revealed a parallelism among the extents of phospholipidosis, apoptosis, and proliferative response for three aminoglycosides (gentamicin >> amikacin congruent with isepamicin). By contrast, netilmicin induced a marked phospholipidosis but a moderate apoptosis and proliferative response. We conclude that rats treated with gentamicin develop an apoptotic process as part of the various cortical alterations induced by this antibiotic at low doses. Netilmicin, and still more amikacin and isepamicin, appears safer in this respect. Whereas a relation between aminoglycoside-induced tubular apoptosis and cortical proliferative response seems to be established, no simple correlation with phospholipidosis can be drawn. PMID- 10681337 TI - Flow cytometric monitoring of antibiotic-induced injury in Escherichia coli using cell-impermeant fluorescent probes. AB - Three fluorescent nucleic acid binding dyes-propidium iodide, TO-PRO-1, and SYTOX green-were evaluated, and their abilities to distinguish between bacterial cells with and without an intact cytoplasmic membrane were compared. Each dye was readily able to discriminate between healthy and permeabilized cells of Escherichia coli, although SYTOX green showed a greater enhancement in fluorescence intensity on staining-compromised, as opposed to healthy, cells in log-phase growth, than either PI or TO-PRO-1. Flow cytometric analysis of E. coli stained with these dyes after exposing them to several antimicrobial agents showed that all three dyes were able to detect antimicrobial action. Notably, however, the intensity of the cell-associated fluorescence was related to the mechanism of action of the antimicrobial agent. Large changes in fluorescence intensity were observed for all the dyes subsequent to beta-lactam antibiotic action, but smaller changes (or no change) were seen subsequent to exposure to antimicrobials acting directly or indirectly on nucleic acid synthesis. Furthermore, cell-associated fluorescence did not relate to loss of viability as determined by plate counts. Despite offering much insight into antimicrobial mechanisms of action, these fundamental problems become relevant to the development of rapid antimicrobial susceptibility tests if colony formation is used as the standard. PMID- 10681338 TI - Flow cytometric investigation of filamentation, membrane patency, and membrane potential in Escherichia coli following ciprofloxacin exposure. AB - Ninety-eight percent of the cells in a population of Escherichia coli in log phase growth lost colony-forming ability after being exposed for 3 h to the quinolone antibiotic ciprofloxacin at four times the MIC in nutrient broth, a concentration easily reached in vivo. Flow cytometric analysis, however, demonstrated that only 68% of this bacterial population had lost membrane potential, as judged by the membrane potential-sensitive dye bis-(1,3 dibutylbarbituric acid) trimethine oxonol [DiBAC(4)(3)], and only 30% could no longer exclude the nucleic acid-binding dye propidium iodide (PI), reflecting lost membrane integrity, efflux mechanisms, or both. Subsequent removal of ciprofloxacin and resuspension in nutrient broth resulted in renewed cell division after 2 h, with a calculated postantibiotic effect (PAE) time of 57 min. The proportion of DiBAC- and PI-fluorescent cells in this recovering population remained stable for more than 4 h after antibiotic removal. Eighty percent of cells present at drug removal were filamentous. Their number subsequently decreased with time, and the increase in particle count seen at the end of the PAE resulted from the division of short cells. Exposure to ciprofloxacin in the presence of the protein synthesis inhibitor chloramphenicol increased colony forming ability to 60% of starting population numbers. In contrast to ciprofloxacin alone, this antibiotic combination resulted in insignificant filamentation and no dye uptake. Subsequent drug removal and resuspension in nutrient broth resulted in the appearance of filaments within 1 h, with 69% of the population forming filaments at 3 h. Dye uptake was also seen, with 20% of the population fluorescing with either dye after 4 h. We were unable to relate dye uptake to the viable count. Cell division resumed 240 min after removal of both drugs, yielding a PAE calculated at 186 min. Inhibition of protein synthesis with chloramphenicol prevented ciprofloxacin-induced changes in bacterial morphology, cell membrane potential, and ability to exclude nucleic acid-binding dye. These changes persisted beyond the end of the classically defined PAE and were not a definite indicator of cell death as defined by loss of colony formation, which related at least in part to filamentation. PMID- 10681339 TI - A standardized plaque reduction assay for determination of drug susceptibilities of cytomegalovirus clinical isolates. AB - Twelve laboratories collaborated in formulating and testing a standardized plaque reduction assay for cytomegalovirus (CMV) cell-associated clinical isolates. Four characterized and plaque-purified CMV strains, as well as six coded clinical isolates obtained after antiviral therapy, were distributed and tested. Good agreement was obtained for four of the clinical isolates, but a broad distribution of results was obtained for two isolates. Analysis of these results indicates the problems associated with clinical isolates, including the large genetic variability and the highly cell-associated phenotype. This collaborative effort, by addressing these problems, represents a significant step toward the development of a standardized assay. PMID- 10681340 TI - A double-blind placebo-controlled study of an infusion of lexipafant (Platelet activating factor receptor antagonist) in patients with severe sepsis. AB - Platelet-activating factor (PAF) is a potent endogenous proinflammatory mediator implicated in the pathogenesis of septic shock. A double-blind randomized placebo controlled trial of an intravenous PAF receptor antagonist (lexipafant) was conducted with 131 adult Thai patients with suspected severe sepsis (66 of whom had positive blood cultures). Detailed serial clinical, biochemical, and cytokine measurements were performed. Lexipafant treatment was well tolerated. The 28-day mortality in the lexipafant group (61.4%) was similar to that in the placebo group (62.6%). There was also no evidence that lexipafant affected clinical or biochemical measures of disease severity or the profile of sequentially measured plasma cytokine levels. PAF may not have an important role in the pathogenesis of severe sepsis. PMID- 10681341 TI - Pharmacokinetics and pharmacodynamics of lumefantrine (benflumetol) in acute falciparum malaria. AB - The objective of this study was to conduct a prospective population pharmacokinetic and pharmacodynamic evaluation of lumefantrine during blinded comparisons of artemether-lumefantrine treatment regimens in uncomplicated multidrug-resistant falciparum malaria. Three combination regimens containing an average adult lumefantrine dose of 1,920 mg over 3 days (four doses) (regimen A) or 2,780 mg over 3 or 5 days (six doses) (regimen B or C, respectively) were given to 266 Thai patients. Detailed observations were obtained for 51 hospitalized adults, and sparse data were collected for 215 patients of all ages in a community setting. The population absorption half-life of lumefantrine was 4.5 h. The model-based median (5th and 95th percentiles) peak plasma lumefantrine concentrations were 6.2 (0.25 and 14.8) microgram/ml after regimen A, 9. 0 (1.1 and 19.8) microgram/ml after regimen B, and 8 (1.4 and 17.4) microgram/ml after regimen C. During acute malaria, there was marked variability in the fraction of drug absorbed by patients (coefficient of variation, 150%). The fraction increased considerably and variability fell with clinical recovery, largely because food intake was resumed; taking a normal meal close to drug administration increased oral bioavailability by 108% (90% confidence interval, 64 to 164) (P, 0.0001). The higher-dose regimens (B and C) gave 60 and 100% higher areas under the concentration-time curves (AUC), respectively, and thus longer durations for which plasma lumefantrine concentrations exceeded the putative in vivo MIC of 280 microgram/ml (median for regimen B, 252 h; that for regimen C, 298 h; that for regimen A, 204 h [P, 0.0001]) and higher cure rates. Lumefantrine oral bioavailability is very dependent on food and is consequently poor in acute malaria but improves markedly with recovery. The high cure rates with the two six-dose regimens resulted from increased AUC and increased time at which lumefantrine concentrations were above the in vivo MIC. PMID- 10681342 TI - The biopesticide Paenibacillus popilliae has a vancomycin resistance gene cluster homologous to the enterococcal VanA vancomycin resistance gene cluster. AB - We have previously identified, in Paenibacillus popilliae, a 708-bp sequence which has homology to the sequence of the enterococcal vanA gene. We have performed further studies revealing five genes encoding homologues of VanY, VanZ, VanH, VanA, and VanX in P. popilliae. The predicted amino acid sequences are similar to those in VanA vancomycin-resistant enterococci: 61% identity for VanY, 21% for VanZ, 74% for VanH, 77% for VanA, and 79% for VanX. The genes in P. popilliae may have been a precursor to or have had ancestral genes in common with vancomycin resistance genes in enterococci. The use of P. popilliae biopesticidal preparations in agricultural practice may have an impact on bacterial resistance in human pathogens. PMID- 10681343 TI - Molecular mechanisms of fluoroquinolone resistance in Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - Twenty P. aeruginosa isolates were collected from six cystic fibrosis (CF) patients, aged 27 to 33, in 1994 (9 isolates) and 1997 (11 isolates) at the CF Center, Copenhagen, Denmark, and were typed by pulse-field gel electrophoresis (PFGE) or ribotyping. Five of the patients had isolates with the same PFGE or ribotyping patterns in 1997 as in 1994, and ciprofloxacin had a two- to fourfold higher MIC for the isolates collected in 1997 than those from 1994. Genomic DNA was amplified for gyrA, parC, mexR, and nfxB by PCR and sequenced. Eleven isolates had mutations in gyrA, seven isolates had mutations at codon 83 (Thr to Ile), and four isolates had mutations at codon 87 (Asp to Asn or Tyr). Sixteen isolates had mutations in nfxB at codon 82 (Arg to Leu). Increased amounts of OprN were found in six isolates and OprJ in eight isolates as determined by immunoblotting. No isolates had mutations in parC or mexR. Six isolates had mutations in efflux pumps without gyrA mutations. The average number of mutations was higher in isolates from 1997 than in those from 1994. The results also suggested that efflux resistance mechanisms are more common in isolates from CF patients than in strains from urine and wounds from non-CF patients, in which mutations in gyrA and parC dominate (S. Jalal and B. Wretlind, Microb. Drug Resist. 4:257-261, 1998). PMID- 10681344 TI - Effects of atovaquone and diospyrin-based drugs on the cellular ATP of Pneumocystis carinii f. sp. carinii. AB - Atovaquone (also called Mepron, or 566C80) is a napthoquinone used for the treatment of infections caused by pathogens such as Plasmodium spp. and Pneumocystis carinii. The mechanism of action against the malarial parasite is the inhibition of dihydroorotate dehydrogenase (DHOD), a consequence of blocking electron transport by the drug. As an analog of ubiquinone (coenzyme Q [CoQ]), atovaquone irreversibly binds to the mitochondrial cytochrome bc(1) complex; thus, electrons are not able to pass from dehydrogenase enzymes via CoQ to cytochrome c. Since DHOD is a critical enzyme in pyrimidine biosynthesis, and because the parasite cannot scavenge host pyrimidines, the drug is lethal to the organism. Oxygen consumption in P. carinii is inhibited by the drug; thus, electron transport has also been identified as the drug target in P. carinii. However, unlike Plasmodium DHOD, P. carinii DHOD is inhibited only at high atovaquone concentrations, suggesting that the organism may salvage host pyrimidines and that atovaquone exerts its primary effects on ATP biosynthesis. In the present study, the effect of atovaquone on ATP levels in P. carinii was measured directly from 1 to 6 h and then after 24, 48, and 72 h of exposure. The average 50% inhibitory concentration after 24 to 72 h of exposure was 1.5 microgram/ml (4.2 microM). The kinetics of ATP depletion were in contrast to those of another family of naphthoquinone compounds, diospyrin and two of its derivatives. Whereas atovaquone reduced ATP levels within 1 h of exposure, the diospyrins required at least 48 h. After 72 h, the diospyrins were able to decrease ATP levels of P. carinii at nanomolar concentrations. These data indicate that although naphthoquinones inhibit the electron transport chain, the molecular targets in a given organism are likely to be distinct among members of this class of compounds. PMID- 10681345 TI - Enzymes catalyzing the early steps of clavulanic acid biosynthesis are encoded by two sets of paralogous genes in Streptomyces clavuligerus. AB - Genes encoding the proteins required for clavulanic acid biosynthesis and for cephamycin biosynthesis are grouped into a "supercluster" in Streptomyces clavuligerus. Nine open reading frames (ORFs) associated with clavulanic acid biosynthesis were located in a 15-kb segment of the supercluster, including six ORFs encoding known biosynthetic enzymes or regulatory proteins, two ORFs that have been reported previously but whose involvement in clavulanic acid biosynthesis is unclear, and one ORF not previously reported. Evidence for the involvement of these ORFs in clavulanic acid production was obtained by generating mutants and showing that all were defective for clavulanic acid production when grown on starch asparagine medium. However, when five of the nine mutants, including mutants defective in known clavulanic acid biosynthetic enzymes, were grown in a soy-based medium, clavulanic acid-producing ability was restored. This ability to produce clavulanic acid when seemingly essential biosynthetic enzymes have been mutated suggests that paralogous genes encoding functionally equivalent proteins exist for each of the five genes but that these paralogues are expressed only in the soy-based medium. The five genes that have paralogues encode proteins involved in the early steps of the pathway common to the biosynthesis of both clavulanic acid and the other clavam metabolites produced by this organism. No evidence was seen for paralogues of the four remaining genes involved in late, clavulanic acid-specific steps in the pathway. PMID- 10681346 TI - Pharmacokinetics of SCH 56592, a new azole broad-spectrum antifungal agent, in mice, rats, rabbits, dogs, and cynomolgus monkeys. AB - SCH 56592 is a new broad-spectrum azole antifungal agent that is in phase 3 clinical trials for the treatment of serious systemic fungal infections. The pharmacokinetics of this drug candidate were evaluated following its intravenous (i.v.) or oral (p.o.) administration as a solution in hydroxypropyl-beta cyclodextrin (HPbetaCD) or oral administration as a suspension in 0.4% methylcellulose (MC) in studies involving mice, rats, rabbits, dogs, and cynomolgus monkeys. SCH 56592 was orally bioavailable in all species. The oral bioavailability was higher with the HPbetaCD solution (range, 52 to approximately 100%) than from the MC suspension (range, 14 to 48%) and was higher in mice ( approximately 100% [HPbetaCD] and 47% [MC]), rats ( approximately 66% [HPbetaCD] and 48% [MC]), and dogs (72% [HPbetaCD] and 37% [MC]) than in monkeys (52% [HPbetaCD] and 14% [MC]). In rabbits, high concentrations in serum suggested good oral bioavailability with the MC suspension. The i.v. terminal-phase half-lives were 7 h in mice and rats, 15 h in dogs, and 23 h in monkeys. In rabbits, the oral half-life was 9 h. In species given increasing oral doses (mice, rats, and dogs), serum drug concentrations were dose related. Food produced a fourfold increase in serum drug concentrations in dogs. Multiple daily doses of 40 mg of SCH 56592/kg of body weight for eight consecutive days to fed dogs resulted in higher concentrations in serum, indicating accumulation upon multiple dosing, with an accumulation index of approximately 2.6. Concentrations above the MICs and minimum fungicidal concentrations for most organisms were observed at 24 h following a single oral dose in MC suspension in all five species studied (20 mg/kg for mice, rats, and rabbits and 10 mg/kg for dogs and monkeys), suggesting that once-daily administration of SCH 56592 in human subjects would be a therapeutically effective dosage regimen. PMID- 10681347 TI - Mutations in ribosomal protein L16 conferring reduced susceptibility to evernimicin (SCH27899): implications for mechanism of action. AB - A clinical isolate of Streptococcus pneumoniae (SP#5) that showed decreased susceptibility to evernimicin (MIC, 1.5 microgram/ml) was investigated. A 4,255 bp EcoRI fragment cloned from SP#5 was identified by its ability to transform evernimicin-susceptible S. pneumoniae R6 (MIC, 0.03 microgram/ml) such that the evernimicin MIC was 1.5 microgram/ml. Nucleotide sequence analysis of this fragment revealed that it contained portions of the S10-spc ribosomal protein operons. The nucleotide sequences of resistant and susceptible isolates were compared, and a point mutation (thymine to guanine) that causes an Ile52-Ser substitution in ribosomal protein L16 was identified. The role of this mutation in decreasing susceptibility to evernimicin was confirmed by direct transformation of the altered L16 gene. The presence of the L16 mutation in the resistant strain suggests that evernimicin is an inhibitor of protein synthesis. This was confirmed by inhibition studies using radiolabeled substrates, which showed that the addition of evernimicin at sub-MIC levels resulted in a rapid decrease in the incorporation of radiolabeled isoleucine in a susceptible isolate (SP#3) but was much less effective against SP#5. The incorporation of isoleucine showed a linear response to the dose level of evernimicin. The incorporation of other classes of labeled substrates was unaffected or much delayed, indicating that these were secondary effects. PMID- 10681348 TI - Synergistic antifungal activities of bafilomycin A(1), fluconazole, and the pneumocandin MK-0991/caspofungin acetate (L-743,873) with calcineurin inhibitors FK506 and L-685,818 against Cryptococcus neoformans. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that causes life threatening infections of the central nervous system. Existing therapies include amphotericin B, fluconazole, and flucytosine, which are limited by toxic side effects and the emergence of drug resistance. We recently demonstrated that the protein phosphatase calcineurin is required for growth at 37 degrees C and virulence of C. neoformans. Because calcineurin is the target of potent inhibitors in widespread clinical use, cyclosporine and FK506 (tacrolimus), it is an attractive drug target for novel antifungal agents. Here we have explored the synergistic potential of combining the calcineurin inhibitor FK506 or its nonimmunosuppressive analog, L-685,818, with other antifungal agents and examined the molecular basis of FK506 action by using genetically engineered fungal strains that lack the FK506 target proteins FKBP12 and calcineurin. We demonstrate that FK506 exhibits marked synergistic activity with the H(+)ATPase inhibitor bafilomycin A(1) via a novel action distinct from calcineurin loss of function. FK506 also exhibits synergistic activity with the pneumocandin MK 0991/caspofungin acetate (formerly L-743,873), which targets the essential beta 1,3 glucan synthase, and in this case, FK506 action is mediated via FKBP12 dependent inhibition of calcineurin. Finally, we demonstrate that FK506 and fluconazole have synergistic activity that is independent of both FKBP12 and calcineurin and may involve the known ability of FK506 to inhibit multidrug resistance pumps, which are known to export azoles from fungal cells. In summary, our studies illustrate the potential for synergistic activity of a variety of different drug combinations and the power of molecular genetics to define the mechanisms of drug action, as well as identify a novel action of FK506 that could have profound implications for therapeutic or toxic effects in other organisms, including humans. PMID- 10681349 TI - Bloodstream infections due to Candida species: SENTRY antimicrobial surveillance program in North America and Latin America, 1997-1998. AB - An international program of surveillance of bloodstream infections (BSI) in the United States, Canada, and Latin America detected 306 episodes of candidemia in 34 medical centers (22 in the United States, 6 in Canada, and 6 in Latin America) in 1997 and 328 episodes in 34 medical centers (22 in the United States, 5 in Canada, and 7 in Latin America) in 1998. Of the 634 BSI, 54.3% were due to Candida albicans, 16.4% were due to C. glabrata, 14.9% were due to C. parapsilosis, 8.2% were due to C. tropicalis, 1.6% were due to C. krusei, and 4.6% were due to other Candida spp. The percentage of BSI due to C. albicans decreased very slightly in the United States between 1997 and 1998 (56.2 to 54.4%; P = 0.68) and increased in both Canada (52.6 to 70.1%; P = 0.05) and Latin America (40.5 to 44. 6%; P = 0.67). C. glabrata was the second most common species observed overall, and the percentage of BSI due to C. glabrata increased in all three geographic areas between 1997 and 1998. C. parapsilosis was the third most prevalent BSI isolate in both Canada and Latin America, accounting for 7.0 and 18.5% of BSI, respectively. Resistance to fluconazole (MIC, >/=64 microgram/ml) and itraconazole (MIC, >/=1.0 microgram/ml) was observed infrequently in both 1997 (2.3 and 8.5%, respectively) and 1998 (1.5 and 7.6%, respectively). Among the different species of Candida, resistance to fluconazole and itraconazole was observed in C. glabrata and C. krusei, whereas isolates of C. albicans, C. parapsilosis, and C. tropicalis were all highly susceptible to both fluconazole (98.9 to 100% susceptible) and itraconazole (96.4 to 100% susceptible). Isolates from Canada and Latin America were generally more susceptible to both triazoles than U.S. isolates were. Continued surveillance appears necessary to detect these important changes. PMID- 10681350 TI - Indinavir pharmacokinetics and parmacodynamics in children with human immunodeficiency virus infection. AB - The indinavir dosage regimen currently used for human immunodeficiency virus (HIV)-infected children is not based on pharmacokinetic data obtained in the target patient population. The purpose of our study was to characterize indinavir pharmacokinetics and pharmacodynamics in HIV-infected children. Eleven children (age range, 9.0 to 13.6 years; weight range, 21.7 to 56.0 kg) receiving indinavir (500 mg/m(2) every 8 h) in combination with lamivudine and stavudine were studied. The correlation of indinavir pharmacokinetic parameters and demographic parameters was evaluated. Also, the pharmacodynamic relationship between parameters of indinavir exposure and parameters of renal toxicity and immunologic recovery was studied. The area under the indinavir concentration-time curve (AUC) and patient body surface area (BSA) showed a significant negative correlation (r = 0.73; P = 0.012). Patients with smaller BSA had excessive indinavir AUC compared to adults. On the other hand, the median minimum drug concentration in plasma (C(min)) was lower than that reported for adults. The maximum indinavir concentration in serum was higher in patients with renal toxicity (5 out of 11 children), but the difference was not statistically significant (15.3 +/- 8.2 versus 9.8 +/- 4.4 mg/liter; P = 0.19). There was a trend toward higher immunologic efficacy in patients with greater indinavir exposure: the time averaged AUC of the percentage of CD4(+) lymphocytes over the baseline value for patients with indinavir C(min) > 95% inhibitory concentration (IC(95)) was higher than in patients with C(min) < IC(95) (P = 0. 068). Our study suggests that a dose reduction may be appropriate for children with small BSA and that a 6-h dosage regimen may be indicated for a substantial percentage of patients. Due to the low number of patients enrolled in this study, our results should be confirmed by a larger study. PMID- 10681351 TI - Intravitreal, retinal, and central nervous system foscarnet concentrations after rapid intravenous administration to rabbits. AB - Retinal, vitreous humor, brain, and cerebrospinal fluid (CSF) foscarnet levels were measured by high-performance liquid chromatography after administration of an intravenous dose of 120 mg/kg of body weight to 32 pigmented rabbits. A pharmacokinetic analysis was done using a two-compartment model. The penetration ratios, defined as ratios of retinal, vitreous humor, brain, and CSF areas under the concentration-time curve from 0 to 2 h were 110% +/- 1%, 12.3% +/- 0.7%, 118% +/- 1%, and 20.2% +/- 2.2%, respectively. These results suggest a good penetration of foscarnet into the retinal and brain tissues, reaching higher concentrations than those estimated from vitreous humor and CSF levels. PMID- 10681352 TI - Capnocytophaga ochracea: characterization of a plasmid-encoded extended-spectrum TEM-17 beta-lactamase in the phylum Flavobacter-bacteroides. AB - A plasmid-encoded extended-spectrum TEM beta-lactamase with a pI of 5.5 was detected in a Capnocytophaga ochracea clinical isolate. The bla gene was associated with a strong TEM-2 promoter and was derived from bla(TEM-1a) with a single-amino-acid substitution: Glu(104)-->Lys, previously assigned to TEM-17, which is thus the first TEM beta-lactamase to be reported in the phylum Flavobacter-Bacteroides. PMID- 10681353 TI - Iron-chelating activity of tetracyclines and its impact on the susceptibility of Actinobacillus actinomycetemcomitans to these antibiotics. AB - Three tetracyclines (tetracycline, doxycycline, and minocycline) were found to possess iron-chelating activity in a colorimetric siderophore assay. Determination of MICs indicated that the activity of doxycycline against the periodontopathogen Actinobacillus actinomycetemcomitans was only slightly influenced by the presence of an excess of iron that likely saturates the antibiotic. On the other hand, the MICs of doxycycline and minocycline were significantly lower for A. actinomycetemcomitans cultivated under iron-poor conditions than under iron-rich conditions. PMID- 10681354 TI - Gemifloxacin is effective in experimental pneumococcal meningitis. AB - In a rabbit model of Streptococcus pneumoniae meningitis, 5 mg of gemifloxacin mesylate (SB-265805) per kg/h reduced the bacterial titers in cerebrospinal fluid (CSF) almost as rapidly as 10 mg of ceftriaxone per kg/h (Deltalog CFU/ml/h +/- standard deviation [SD], -0.25 +/- 0.09 versus -0.38 +/- 0.11; serum and CSF concentrations of gemifloxacin were 2.1 +/- 1.4 mg/liter and 0.59 +/- 0.38 mg/liter, respectively, at 24 h). Coadministration of 1 mg of dexamethasone per kg did not affect gemifloxacin serum and CSF levels (2.7 +/- 1.4 mg/liter and 0.75 +/- 0.34 mg/liter, respectively, at 24 h) or activity (Deltalog CFU/ml/h +/- SD, -0.26 +/- 0.11). PMID- 10681355 TI - Pulsed ultrasound enhances the killing of Escherichia coli biofilms by aminoglycoside antibiotics in vivo. AB - Escherichia coli biofilms on two polyethylene disks were implanted subcutaneously into rabbits receiving systemic gentamicin. Ultrasound was applied for 24 h to one disk. Both disks were removed, and viable bacteria were counted. Pulsed ultrasound significantly reduced bacterial viability below that of nontreated biofilms without damage to the skin. PMID- 10681356 TI - Comparative activities of ciprofloxacin and levofloxacin against Streptococcus pneumoniae in an In vitro dynamic model. AB - The activities of levofloxacin (500 mg every 24 h) and ciprofloxacin (750 mg every 12 h) against six pneumococcal isolates in an in vitro dynamic model were compared. For one strain, levofloxacin reduced the inoculum by over 4 log CFU/ml and ciprofloxacin reduced the inoculum by over 2 log CFU/ml. For four isolates, both drugs reduced inocula by 4 log CFU/ml within 6 h, suggesting that this dose of ciprofloxacin should be as effective as levofloxacin against these pneumococci. PMID- 10681357 TI - Occurrence of the new tetracycline resistance gene tet(W) in bacteria from the human gut. AB - Members of our group recently identified a new tetracycline resistance gene, tet(W), in three genera of rumen obligate anaerobes. Here, we show that tet(W) is also present in bacteria isolated from human feces. The tet(W) genes found in human Fusobacterium prausnitzii and Bifidobacterium longum isolates were more than 99.9% identical to those from a rumen isolate of Butyrivibrio fibrisolvens. PMID- 10681358 TI - Postantibiotic effects and bactericidal activities of clarithromycin-14-hydroxy clarithromycin, versus those of amoxicillin-clavulanate, against anaerobes. AB - The bactericidal activities and postantibiotic effects (PAE) of clarithromycin-14 hydroxy-clarithromycin and amoxicillin-clavulanate against Bacteroides fragilis and Peptostreptococcus anaerobius were determined. A concentration of twice the MIC resulted in bactericidal activity against four of four and three of four organisms at 24 h with clarithromycin-14-hydroxy-clarithromycin and amoxicillin clavulanate, respectively. The PAE of clarithromycin-14-hydroxy-clarithromycin was 1.44 to 3.20 h, compared to the less than 1 h of amoxicillin-clavulanate. Clarithromycin-14-hydroxy-clarithromycin possesses good activity against susceptible anaerobes. PMID- 10681359 TI - Efficacy of SCH56592 in a rabbit model of invasive aspergillosis. AB - SCH56592 (SCH) was evaluated in an immunosuppressed rabbit model of invasive aspergillosis. SCH was more effective than similar doses of itraconazole and as effective as amphotericin B in the clearance of Aspergillus spp. from tissues. Compared with controls, SCH regimens reduced mortality, improved survival, and significantly reduced tissue colony counts. PMID- 10681360 TI - Antiviral activity of 2'-deoxy-3'-oxa-4'-thiocytidine (BCH-10652) against lamivudine-resistant human immunodeficiency virus type 1 in SCID-hu Thy/Liv mice. AB - Oral administration of 2'-deoxy-3'-oxa-4'-thiocytidine (BCH-10652), a nucleoside analog structurally similar to lamivudine (3TC), caused dose-dependent inhibition of viral replication in SCID-hu Thy/Liv mice infected with human immunodeficiency virus type 1 NL4-3 and with an NL4-3 clone containing the M184V mutation in reverse transcriptase that confers resistance to 3TC. These experiments demonstrate the utility of this mouse model for evaluating drug resistance and for performing direct comparisons between antiviral compounds in vivo. PMID- 10681361 TI - Streptococcus gordonii strains resistant to fluorodeoxyuridine contain mutations in the thymidine kinase gene and are deficient in thymidine kinase activity. AB - Mutants of Streptococcus gordonii resistant to 5-fluorodeoxyuridine (FUdR(r)) were isolated. Each strain contained a point mutation resulting in the premature termination of the thymidine kinase (TK) open reading frame (tdk). In vitro translation of the mutant tdk coding regions resulted in synthesis of truncated TK polypeptides deficient in TK activity. PMID- 10681362 TI - Characterization of the Tn916-like transposon Tn3872 in a strain of abiotrophia defectiva (Streptococcus defectivus) causing sequential episodes of endocarditis in a child. AB - Clinical blood isolates from three sequential episodes of endocarditis occurring over a 6-month period in a child with a malformative cardiopathy were investigated. All isolates identified as Abiotrophia defectiva were resistant to erythromycin-clindamycin and to tetracycline-minocycline, due to the presence of sequences homologous to the erythromycin resistance gene ermB and to the tetracycline resistance gene tet(M), respectively. These resistance genes were located on a chromosomally borne composite Tn916-related transposon. These results demonstrate the involvement of conjugative transposons in the dissemination of antibiotic resistance in the genus Abiotrophia. PMID- 10681364 TI - A convenient assay for estimating the possible involvement of efflux of fluoroquinolones by Streptococcus pneumoniae and Staphylococcus aureus: evidence for diminished moxifloxacin, sparfloxacin, and trovafloxacin efflux. AB - We developed a simplified assay for estimating efflux by measuring the effect of reserpine on the growth of Streptococcus pneumoniae and Staphylococcus aureus over 7 h. Reserpine enhanced ciprofloxacin and levofloxacin 17 to 68%. The hydrophobic drug trovafloxacin and the drug moxifloxacin, with a bulky C-7 substituent but hydrophilicity similar to that of levofloxacin, showed little (0 to 11%) reserpine-enhancing effect. The ease of resistant mutant strain selection correlated with efflux susceptibility. PMID- 10681363 TI - Delavirdine susceptibilities and associated reverse transcriptase mutations in human immunodeficiency virus type 1 isolates from patients in a phase I/II trial of delavirdine monotherapy (ACTG 260). AB - The development of human immunodeficiency virus type 1 resistance to delavirdine (DLV) was studied in subjects receiving DLV monotherapy. Phenotypic resistance developed in 28 of 30 subjects within 8 weeks. K103N and Y181C, which confer nonnucleoside reverse transcriptase inhibitor (NNRTI) cross-resistance, were the predominant reverse transcriptase mutations. P236L, which confers DLV resistance but hypersensitivity to other NNRTIs, developed in <10% of isolates. PMID- 10681365 TI - Comparative in vitro activities of ciprofloxacin, gemifloxacin, grepafloxacin, moxifloxacin, ofloxacin, sparfloxacin, trovafloxacin, and other antimicrobial agents against bloodstream isolates of gram-positive cocci. AB - The in vitro activity of gemifloxacin against 316 bloodstream isolates of staphylococci, pneumococci, and enterococci was compared with the activities of six fluoroquinolones and three other antimicrobial agents. Of the antimicrobial agents tested, gemifloxacin was the most potent against penicillin-intermediate and -resistant pneumococci, methicillin-susceptible and -resistant Staphylococcus epidermidis isolates, and coagulase-negative staphylococci. PMID- 10681366 TI - Editorial PMID- 10681367 TI - In vitro-in vivo scaling of CYP kinetic data not consistent with the classical Michaelis-Menten model. AB - Strategies for the prediction of in vivo drug clearance from in vitro drug metabolite kinetic data are well established for the rat. In this animal species, metabolism rate-substrate concentration relationships can commonly be described by the classic hyperbola consistent with the Michaelis-Menten model and simple scaling of the parameter intrinsic clearance (CL(int) - the ratio of V(max) to K(m)) is particularly valuable. The in vitro scaling of kinetic data from human tissue is more complex, particularly as many substrates for cytochrome P450 (CYP) 3A4, the dominant human CYP, show nonhyperbolic metabolism rate-substrate concentration curves. This review critically examines these types of data, which require the adoption of an enzyme model with multiple sites showing cooperative binding for the drug substrate, and considers the constraints this kinetic behavior places on the prediction of in vivo pharmacokinetic characteristics, such as metabolic stability and inhibitory drug interaction potential. The cases of autoactivation and autoinhibition are discussed; the former results in an initial lag in the rate-substrate concentration profile to generate a sigmoidal curve whereas the latter is characterized by a convex curve as V(max) is not maintained at high substrate concentrations. When positive cooperativity occurs, we suggest the use of CL(max), the maximal clearance resulting from autoactivation, as a substitute for CL(int). The impact of heteroactivation on this approach is also of importance. In the case of negative cooperativity, care in using the V(max)/K(m) approach to CL(int) determination must be taken. Examples of substrates displaying each type of kinetic behavior are discussed for various recombinant CYP enzymes, and possible artifactual sources of atypical rate-concentration curves are outlined. Finally, the consequences of ignoring atypical Michaelis-Menten kinetic relationships are examined, and the inconsistencies reported for both different substrates and sources of recombinant CYP3A noted. PMID- 10681368 TI - Bioactivation and covalent binding of hydroxyfluperlapine in human neutrophils: implications for fluperlapine-induced agranulocytosis. AB - The use of fluperlapine and the structurally related clozapine has been associated with the induction of agranulocytosis in humans. Unlike clozapine, fluperlapine is relatively resistant to chemical and biochemical oxidations. In this study we demonstrated that 7-hydroxyfluperlapine, the major metabolite of fluperlapine in humans, is oxidized to a reactive intermediate by HOCl and by myeloperoxidase in the presence of H(2)O(2) and Cl(-). This reactive intermediate was identified as an iminoquinone species with a M + 1 ion at m/z 324 by mass spectrometry. The iminoquinone intermediate was trapped by N-acetyl-L-cysteine (NAC) as well as GSH. NMR spectra of the NAC adducts indicated that the NAC was bound to the 6 and 9 positions of the aromatic ring. This is the same orientation as the binding of nucleophiles to the reactive metabolite of clozapine. We were able to use an antibody against clozapine to demonstrate that 7 hydroxyfluperlapine, but not fluperlapine itself, covalently modifies human myeloperoxidase. Furthermore, we demonstrated that 7-hydroxyfluperlapine is metabolized by activated neutrophils to a reactive intermediate that covalently binds to neutrophils. In the presence of NAC or GSH, such covalent binding was inhibited and the NAC or GSH adducts were formed. Thus, the reactivity and even the orientation of the binding of the reactive metabolite of 7 hydroxyfluperlapine is very similar to that of clozapine. These results provide a mechanism for the formation of a reactive metabolite of fluperlapine similar to clozapine that may explain its ability to induce agranulocytosis. PMID- 10681369 TI - The role of mdr1a P-glycoprotein in the biliary and intestinal secretion of doxorubicin and vinblastine in mice. AB - Drug-transporting P-glycoproteins are abundantly present in the liver and the intestinal wall. We have now investigated their role in the biliary and intestinal secretion of the anticancer drugs doxorubicin (unlabeled: 5 mg/kg) and vinblastine ((3)H-labeled: 1 mg/kg) i.v. administered to wild-type and mdr1a P glycoprotein knockout [mdr1a(-/-)] mice. At 90 min after drug administration, levels of unchanged drug and metabolites in plasma, intestinal contents, and bile were determined by high-performance liquid chromatography and radioactivity by liquid scintillation counting. The bile of both wild-type and mdr1a(-/-) mice contained only minor amounts of unchanged vinblastine, whereas the total biliary secretion of unknown (3)H-labeled breakdown products was about 25 to 30% of the dose. The direct secretion of unchanged vinblastine through the gut wall was 6.7 and 3.3% of the dose in wild-type and mdr1a(-/-) mice, respectively. The biliary secretion of unchanged doxorubicin decreased from 13.3% of the dose to only 2.4% in the absence of mdr1a P-glycoprotein. Approximately 10% of the dose was secreted as unchanged doxorubicin into the intestinal contents of both types of mice. Thus, the absence of mdr1a P-glycoprotein affects the fate of vinblastine chiefly by diminishing secretion into the lumen of the small intestine, whereas it affects the fate of doxorubicin chiefly by diminishing secretion of parent drug into bile. PMID- 10681370 TI - The glucocorticoid receptor is essential for induction of cytochrome P-4502B by steroids but not for drug or steroid induction of CYP3A or P-450 reductase in mouse liver. AB - Cytochrome P-4503A, CYP2B, and P-450 reductase are induced by glucocorticoids, antiglucocorticoids such as pregnenolone 16alpha-carbonitrile, and drugs such as rifampin and phenobarbital. Although the pregnane X receptor is reported to mediate steroid and drug activation of CYP3A via a conserved cis-element in CYP3A genes, discrepancies exist between the induction of the endogenous CYP3A genes and the activation of the pregnane X receptor. It is a formal possibility that the glucocorticoid receptor may account for some of these discrepancies. To determine the requirement in vivo of the glucocorticoid receptor in expression of CYP3A and CYP2B, we compared the induction of these proteins in the livers of normal mice and mice with a targeted mutation in the glucocorticoid receptor. Mice lacking the glucocorticoid receptor show no difference in constitutive hepatic expression of CYP3A but show a decrease in the level of CYP2B. Glucocorticoid receptor-deficient mice challenged with either dexamethasone or pregnenolone 16alpha-carbonitrile failed to induce CYP2B proteins, whereas CYP2B was readily induced in (+/+) mice. In contrast, CYP3A and P-450 reductase proteins were induced by either inducer in wild-type and glucocorticoid receptor null mice. Similarly, rifampin induced CYP3A in either wild-type or glucocorticoid receptor-null mice. Despite reports that rifampin is a nonsteroidal ligand for the human glucocorticoid receptor, rifampin failed to induce tyrosine aminotransferase in mice regardless of glucocorticoid receptor genotype, and rifampin did not compete for ligand binding to either mouse or human glucocorticoid receptor. Phenobarbital induced CYP3A, CYP2B, and P-450 reductase in all mice, but the amplitude of induction was diminished 37% in glucocorticoid receptor-null mice. Thus, there are distinctly different essential requirements of CYP3A, CYP2B, and P-450 reductase genes for the glucocorticoid receptor in their induction by steroids and drugs. PMID- 10681371 TI - Diphenhydramine disposition in the sheep maternal-placental-fetal unit: gestational age, plasma drug protein binding, and umbilical blood flow effects on clearance. AB - The objective of this study was to examine the interrelationships between maternal and fetal plasma drug protein binding, umbilical blood flow (Q(um)), gestational age (GA), and maternal-fetal diphenhydramine (DPHM) clearances in chronically instrumented pregnant sheep. Maternal and fetal DPHM placental (CL(mf) and CL(fm), respectively) and nonplacental (CL(mo) and CL(fo), respectively) clearances and steady-state plasma protein binding were determined in 18 pregnant sheep at 124 to 140 days' gestation (term, approximately 145 days). The data demonstrated a highly significant fall of approximately 66% in CL(fm) and a decreasing trend in CL(fo) ( approximately 47%) over the GA range studied. However, no such relationships existed between GA and CL(mf) or CL(mo). Concomitant with this was a decrease in fetal DPHM plasma unbound fraction with GA, with no such change being evident in the mother. Both CL(mo) and CL(fo) were related to the respective DPHM plasma unbound fraction. A strong relationship also existed between fetal plasma unbound fraction and CL(fm). Thus, the decrease in fetal unbound fraction of DPHM during gestation could contribute to the fall in CL(fm), and possibly CL(fo). However, over the GA range studied, fetal DPHM free fraction decreased by approximately 47%, whereas CL(fm) fell by approximately 66%. Because fetal unbound fraction and CL(fm) are linearly related, the GA-associated fall in unbound fraction appears to be insufficient to account for the entire decline in CL(fm). In separate studies in pregnant sheep, we observed a approximately 40% fall in weight-normalized Q(um) between 125 and 137 days' gestation. Because CL(fm) for DPHM is similar to that of flow-limited compounds (e.g., ethanol, antipyrine), this decrease in Q(um) may also contribute to the GA-related fall in CL(fm). PMID- 10681372 TI - Incorporation and retention of radiolabeled S-(+)-and R-(-)-methamphetamine and S(+)- and R(-)-N-(n-butyl)-amphetamine in mouse hair after systemic administration. AB - We examined the incorporation of unlabeled and tritiated enantiomers of methamphetamine (MA) and a more lipophilic analog N-(n-butyl)-amphetamine (BA) into the hair of pigmented (C57) and nonpigmented (Balb/C) mice after systemic administration. We also compared the ability of extraction methods to remove unlabeled and tritiated MA and BA enantiomers from the hair. R(-)-MA, S(+)-MA, [(3)H]R(-)-MA, [(3)H]S(+)-MA, R(-)-BA, S(+)-BA, [(3)H]R-(-)-BA, and [(3)H]S-(+) BA were each administered to C57 and Balb/C mice (23 days of age) by i.p. injection at 8.8 mg/kg daily for 3 days. At 44 days of age, hair samples from the animals were treated with a brief methanol wash, a 24-h extraction with pH 6 phosphate buffer, and a final digestion in 1 N NaOH to free residual drugs from the hair. Labeled drugs in the extracts were quantitated by liquid scintillation counting. Unlabeled drugs were quantitated by gas chromatography/mass spectrometry (GC/MS). GC/MS analysis demonstrated MA and BA to be the major (>90%) species present in the blood during the 24 h after administration. Less than 10% of the MA was N-demethylated. No p-hydroxylated metabolites were found. Blood concentrations of tritiated MA and BA enantiomers measured by liquid scintillation counting agreed well with blood concentrations of unlabeled enantiomers measured by GC/MS. Hair concentrations of S(+)-MA were greater than those of R(-)-MA in both mouse strains, paralleling blood concentrations. There were no enantiomeric differences seen with BA hair accumulation in either strain of mouse. Significantly more MA and BA enantiomers were deposited in pigmented than in nonpigmented hair. With labeled and unlabeled compounds, approximately 30% of S(+)-MA and 60% of R(-)-MA in pigmented hair could be removed by a phosphate extraction. A significant amount of drug could not be removed from the hair by extraction. Greater amounts of drug could be extracted from nonpigmented hair than pigmented. Extracted and residual MA and BA concentrations in pigmented hair were significantly greater when labeled compounds were quantitated by liquid scintillation counting than when unlabeled compounds were quantitated by GC/MS. However, radiotracer and unlabeled drug concentrations were the same in nonpigmented hair. The results demonstrate that hair pigmentation is an important determinant in MA and BA deposition, and that MA and BA deposition is not enantioselective. The data demonstrate a significant amount of MA and BA accumulated is not easily amenable to exhaustive aqueous extraction from the hair. The use of tritiated MA and BA enantiomers demonstrates that a significant amount of MA and BA stored in pigmented hair is structurally different from parent MA and BA, perhaps associated with melanin components of hair. PMID- 10681373 TI - Human cytochrome P-450 metabolism of retinals to retinoic acids. AB - Retinoic acids have important pleiotropic biological effects and thus the potential for human cytochrome P-450s (CYPs) to mediate retinoic acid synthesis was investigated. We examined the retinoic acid synthetic activity of human cDNA expressed CYP1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A4+ cytochrome b(5) (b(5)), 3A5, and 4A11, expressed individually in insect cells together with NADPH-P-450 reductase. Only CYP1A1, 1A2, 1B1, and 3A4+b(5) converted all-trans-retinal (20 microM) to all-trans-retinoic acid with turnover numbers of 0.53, 0.18, 0.20, and 0.41 nmol/min/nmol P-450, respectively. With 9 cis-retinal as substrate, CYP1A2 exhibited a turnover number of 1.58 nmol/min/nmol P-450 whereas CYP1A1, 2C19, and 3A4+b(5) had turnover numbers of 0.40, 0.27, and 0.41 nmol/min/nmol P-450, respectively. For CYP3A4 activities with both retinals, b(5) was required. Kinetic analyses revealed that CYP1A1, 1A2, and 3A4+b(5) with all-trans-retinal had apparent K(m) values of 55, 356, and 255 microM, and V(max) values of 2.0, 8.3, and 6.3 nmol/min/nmol P-450, respectively, and with 9-cis-retinal had K(m) values of 77, 91, and 368 microM, and V(max) values of 2.7, 9.7, and 7.6 nmol/min/nmol P-450, respectively. The 9 cis retinoic acid synthetic activity of a group of 12 human liver microsomes correlated only with the CYP1A2 activity (r = 0.96), implicating CYP1A2 in human liver microsomal metabolism of 9-cis- retinal to 9-cis-retinoic acid. These studies have indicated that human CYPs are capable of catalyzing retinal to retinoic acid metabolism, but the physiological relevance of this metabolism is still unclear. PMID- 10681374 TI - Analysis of soy isoflavone conjugation in vitro and in human blood using liquid chromatography-mass spectrometry. AB - Soybean products containing isoflavones are widely consumed in Western and Asian diets for putative health benefits, but adverse effects are also possible. The conjugated forms of isoflavones present in a soy nutritional supplement (predominately acetyl glucosides) and in blood from two human volunteers after consuming the supplement (7- and 4'-glucuronides and sulfates) were identified using liquid chromatography coupled with electrospray/tandem mass spectrometry. Circulating conjugates of genistein and daidzein were quantified using selective enzymatic hydrolysis and deuterated internal standards for liquid chromatography electrospray/mass spectrometry. The levels of isoflavone glucuronides were much greater than the corresponding sulfates or aglycones. The substrate activities of genistein and daidzein were evaluated with recombinant human UDP glucuronosyl transferase (UGT) and sulfotransferase (SULT) by using enzyme kinetics. The SULTs 1A1*2, 1E, and 2A1 catalyzed formation of a single genistein sulfate; however, SULTs 1A2*1 and 1A3 had no observed activity. None of the SULTs showed activity with daidzein. Although several UGTs (1A1, 1A4, 1A6, 1A7, 1A9, and 1A10) catalyzed 7- and 4'-glucuronidation of genistein or daidzein, the UGT 1A10 isoform, which is found in human colon but not liver, was found to be specific for genistein. Glucuronidation of only genistein was observed in human colon microsomes, although nearly equal activity was observed for daidzein in human liver and kidney microsomes. These findings suggest a prominent role for glucuronidation of genistein in the intestine concomitant with absorption, although hepatic glucuronidation of absorbed genistein and daidzein aglycones is also likely. PMID- 10681375 TI - Metabolism and excretion of [(14)C]celecoxib in healthy male volunteers. AB - We determined the disposition of a single 300-mg dose of [(14)C]celecoxib in eight healthy male subjects. The [(14)C]celecoxib was administered as a fine suspension reconstituted in 80 ml of an apple juice/Tween 80/ethanol mixture. Blood and saliva samples were collected at selected time intervals after dosing. All urine and feces were collected on the 10 consecutive days after dose administration. Radioactivity in each sample was determined by liquid scintillation counting or complete oxidation and liquid scintillation counting. Metabolic profiles in plasma, urine, and feces were obtained by HPLC, and metabolites were identified by mass spectrometry and NMR. [(14)C]Celecoxib was well absorbed, reaching peak plasma concentrations within 2 h of dosing. [(14)C]Celecoxib was extensively metabolized, with only 2.56% of the radioactive dose excreted as celecoxib in either urine or feces. The total percentage of administered radioactive dose recovered was 84.8 +/- 4.9%, with 27.1 +/- 2.2% in the urine and 57.6 +/- 7.3% in the feces. The oxidative metabolism of celecoxib involved hydroxylation of celecoxib at the methyl moiety followed by further oxidation of the hydroxyl group to form a carboxylic acid metabolite. The carboxylic acid metabolite of celecoxib was conjugated with glucuronide to form the 1-O-glucuronide. The percentages of the dose excreted in the feces as celecoxib and the carboxylic acid metabolite were 2.56 +/- 1.09 and 54.4 +/- 6.8%, respectively. The majority of the dose excreted in the urine was the carboxylic acid metabolite (18.8 +/- 2.1%); only a small amount was excreted as the acyl glucuronide (1.48 +/- 0.15%). PMID- 10681376 TI - Biosynthesis of all-trans-retinoic acid from all-trans-retinol: catalysis of all trans-retinol oxidation by human P-450 cytochromes. AB - Oxidative conversion of all-trans-retinol (t-ROH) to all-trans-retinal (t-RAL) is recognized as the rate-limiting step for biosynthesis of all-trans-retinoic acid from t-ROH in mammalian hepatic tissues. The purpose of this study was to investigate the role of human cytochrome P-450 (CYP)-dependent monooxygenation in the conversion of t-ROH to t-RAL. Adult human liver microsomes (HLMS) were incubated with t-ROH, and retinoids generated were identified and quantified by liquid chromatography-mass spectroscopy, HPLC, and other methods. HLMS-catalyzed generation of t-RAL from t-ROH was primarily NADPH-dependent and was strongly inhibited by carbon monoxide. Rates of reactions increased linearly with time and concentrations of HLMS, and exhibited classical substrate saturation. These observations strongly indicated that the reaction proceeded via CYP-catalyzed monooxygenation. On the basis of responses to selective chemical inhibitors, isoforms from CYP family 1 and the CYP3A subfamily appeared to be very active. Members of the CYP2C subfamily and CYP2D6 exhibited lesser activities and CYP2A6, CYP2B6, and CYP2E1 were virtually inactive. cDNA-expressed human CYP enzymes (CYP SUPERSOMES) also were used to assess the capacity of individual CYP enzymes to catalyze the reaction. Based on responses to selective chemical inhibitors, specific activities, and levels present in adult human hepatic tissues, CYP1A2 and CYP3A4 strongly appeared to be the major CYP enzymes catalyzing hepatic oxidative conversion of t-ROH to t-RAL in the adult human liver. CYP1A1 and CYP1B1 SUPERSOMES both exhibited exceptionally high activities, and in extrahepatic tissues, these isoforms could play important roles in biosynthesis of all-trans-retinoic acid from t-ROH. PMID- 10681377 TI - Input rate as a major determinant of furosemide pharmacodynamics: influence of fluid replacement and hypoalbuminemia. AB - To investigate how the response to a bolus and an infusion of furosemide is modulated by the rate of fluid replacement and by hypoalbuminemia, rabbits received 5 mg/kg of furosemide as a bolus or infused over 60 min, whereas diuresis was replaced with 13, 121, or 238 ml/h NaCl 0.9%/glucose 5% (50:50). Natriuretic and diuretic efficiencies were greater with the infusion than with the bolus of furosemide. Fluid replacement increased natriuretic and diuretic efficiency of furosemide bolus but only diuretic efficiency of furosemide infusion. Furosemide net fluid depletion reached a plateau when fluid replacement increased beyond 121 ml/h. Repeated plasmapheresis decreased plasma albumin by 30% (P <.05) and increased furosemide unbound fraction (P <.05). Compared with control rabbits, hypoalbuminemia decreased the natriuresis of the bolus (22.7 +/- 1.5-16.6 +/- 1.3 mmol, P <.05) but not that elicited by furosemide infusion (26.2 +/- 1.8 mmol). Given as a bolus, furosemide natriuretic and diuretic response as a function of its urinary rate of excretion exhibited an hyperbolic relationship, and after its infusion a clockwise hysteresis, denoting tolerance. Plasma renin activity was increased by the bolus and the infusion of furosemide, even in the presence of 121 ml/h of fluid replacement. It is concluded that: 1) the increase in natriuretic/diuretic efficiency of the bolus induced by fluid replacement is greater than when furosemide is infused, 2) furosemide net effect does not increase proportionally to fluid replacement, and 3) the infusion of furosemide prevents the hypoalbuminemia-induced decrease in response of furosemide given as a bolus. PMID- 10681378 TI - Interactions of HIV protease inhibitors with a human organic cation transporter in a mammalian expression system. AB - Recently, we cloned a human organic cation transporter, hOCT1, which is expressed primarily in the liver. hOCT1 plays an important role in the cellular uptake and elimination of various xenobiotics including therapeutically important drugs. HIV protease inhibitors are a new class of therapeutic agents. The purpose of this study was to elucidate the interactions of HIV protease inhibitors with hOCT1 and to determine whether hOCT1 is involved in the elimination of these compounds. We studied the interactions of HIV protease inhibitors with hOCT1 in a transiently transfected human cell line, HeLa. Uptake studies were carried out 40 h post transfection using the radiolabeled model organic cation, [(14)C]tetraethylammonium (TEA), under different experimental conditions. In cis inhibition studies, all of the HIV protease inhibitors tested, i.e., indinavir (IC(50) of 62 microM), nelfinavir (IC(50) of 22 microM), ritonavir (IC(50) of 5.2 microM), and saquinavir (IC(50) of 8.3 microM) inhibited TEA uptake in HeLa cells expressing hOCT1. However, none of the HIV protease inhibitors trans-stimulated [(14)C]TEA uptake, suggesting that they are poorly translocated by hOCT1. Nelfinavir, ritonavir, and saquinavir demonstrated an apparent "trans-inhibition" effect. No enhanced uptake of [(14)C]saquinavir was observed in hOCT1 DNA transfected cells versus empty vector-transfected cells. These data suggest that HIV protease inhibitors are potent inhibitors, but poor substrates, of hOCT1. Some HIV protease inhibitors may potently inhibit the uptake and elimination of cationic drugs that are substrates for hOCT1, leading to potential drug-drug interactions. Other transporters, e.g., MDR1 and MRP1, in HIV-targeted cells may control the intracellular concentrations of HIV protease inhibitors. PMID- 10681379 TI - m-hydroxy benzoylecgonine recovery in fetal guinea pigs. AB - Recently, meta-hydroxybenzoylecgonine (m-OH BE) was identified by gas chromatography-mass spectroscopy during quantitative analysis for cocaine. Identification of m-OH BE in addition to the routinely identified benzoylecgonine by gas chromatography-mass spectroscopy confirmatory assays may increase detection of cocaine-exposed infants and decrease false negative results. However, it is not known whether m-OH BE is derived directly from benzoylecgonine or from hydroxylated cocaine, or whether this metabolite is produced in the fetus or transferred across the placenta from the maternal circulation. We quantitated the recovery of cocaine, benzoylecgonine, and m-OH BE from amniotic fluid, fetal meconium, fetal intestine, and maternal urine for up to 4 days after single dose administration of either cocaine or benzoylecgonine to pregnant time-bred guinea pigs. m-OH BE was recovered from meconium after maternal injections of cocaine and benzoylecgonine. There was no significant detection of m-OH BE from amniotic fluid or intestine and minimal recovery from maternal urine after either cocaine or benzoylecgonine administration. Detection of m-OH BE in meconium increased the identification of in utero exposed guinea pigs, and the greatest yield of m-OH BE from meconium occurred later than that observed for cocaine or benzoylecgonine. PMID- 10681380 TI - On the metabolism of the amphetamine-derived antispasmodic drug mebeverine: gas chromatography-mass spectrometry studies on rat liver microsomes and on human urine. AB - We describe gas chromatography-mass spectrometry studies of the metabolism of the antispasmodic drug mebeverine [Duspatal, (MB)]. MB is the veratric acid (VA) ester of 4-?ethyl-[2-(4-methoxyphenyl)-1-methylethyl]amino?butan-1-ol (MB-OH), which is an N-substituted ethylamphetamine derivative. The metabolites were first identified in rat liver microsome incubates and then detected in urine samples of volunteers through the use of electron impact and positive chemical ionization gas chromatography-mass spectrometry. Urinary conjugates were enzymatically cleaved before analysis. The following phase I metabolites of MB could be identified: VA, O-demethyl VA (vanillic and/or isovanillic acid), O-bisdemethyl VA (protocatechuic acid), MB-OH, hydroxy MB-OH, O-demethyl MB-OH, O-demethyl hydroxy MB-OH, N-desethyl MB-OH, N-desethyl-O-demethyl MB-OH, N-de(hydroxybutyl) MB-OH (methoxy-ethylamphetamine), N-de(hydroxybutyl)-O-demethyl MB-OH (hydroxy ethylamphetamine), and N-bisdealkyl MB-OH (p-methoxy-amphetamine, known as the designer drug PMA). The following, partly overlapping metabolic pathways of MB could be postulated: ester hydrolysis, O-demethylation, ring hydroxylation, N deethylation, and N-de(hydroxybutylation). The latter pathway led to ethylamphetamine derivatives and bisdealkylation led to PMA, which are substances of forensic interest. The metabolites containing alcoholic or phenolic hydroxy groups were partly excreted into urine as conjugates. PMID- 10681381 TI - Selective involvement of cytochrome P450 2D subfamily in in vivo 4-hydroxylation of amphetamine in rat. AB - The cytochrome P450 (P450) 2D subfamily catalyzes ring hydroxylation of amphetamines. We tested the hypothesis that P450 2D is selectively involved in amphetamine 4-hydroxylation. Urinary elimination of 4-hydroxyamphetamine and amphetamine was determined in male Sprague-Dawley rats pretreated with P450 inducers and inhibitors. The urinary 24-h metabolic ratio (amphetamine/4 hydroxyamphetamine) was not affected by the inducers 3-methylcholanthrene, isosafrole, phenobarbital, ethanol, pregnenolone-alpha-carbonitrile, and clofibrate. Isosafrole did, however, increase amphetamine elimination along with urine volume. Urinary elimination of 4-hydroxyamphetamine was significantly decreased by, and the metabolic ratio increased by, the inhibitors 1 aminobenzotriazole, CCl(4), quinidine, quinine, and primaquine. Diallyl sulfide and troleandomycin had no effect. In rat liver microsomes primaquine was shown to be an inhibitor of 2D activity. Urine 4-hydroxyamphetamine content correlated strongly (r(2) = 0. 989) with microsomal P450 2D activity in parallel-treated rats. These studies also substantiate that 4-hydroxylation of amphetamine is selectively performed by the P450 2D subfamily in the rat. PMID- 10681382 TI - CYP2C19 participates in tolbutamide hydroxylation by human liver microsomes. AB - Tolbutamide is a sulfonylurea-type oral hypoglycemic agent whose action is terminated by hydroxylation of the tolylsulfonyl methyl moiety catalyzed by cytochrome P-450 (CYP) enzymes of the human CYP2C subfamily. Although most studies have implicated CYP2C9 as the exclusive catalyst of hepatic tolbutamide hydroxylation in humans, there is evidence that other CYP2C enzymes (e.g., CYP2C19) may also participate. To that end, we used an immunochemical approach to assess the role of individual CYP2Cs in microsomal tolbutamide metabolism. Polyclonal antibodies were raised to CYP2C9 purified from human liver, and were then back-adsorbed against recombinant CYP2C19 coupled to a solid-phase support. Western blotting revealed that the absorbed anti-human CYP2C9 preparation reacted with only recombinant CYP2C9 and the corresponding native protein in hepatic microsomes, and no longer recognized CYP2C19 and CYP2C8. Monospecific anti-CYP2C9 not only retained the ability to inhibit CYP2C9-catalyzed reactions, as evidenced by its marked (90%) inhibition of diclofenac 4'-hydroxylation by purified CYP2C9 and by human liver microsomes, but also exhibited metabolic specificity, as indicated by its negligible (<15%) inhibitory effect on S-mephenytoin 4' hydroxylation by purified CYP2C19 or hepatic microsomes containing CYP2C19. Monospecific anti-CYP2C9 was also found to inhibit rates of tolbutamide hydroxylation by 93 +/- 4 and 78 +/- 6% in CYP2C19-deficient and CYP2C19 containing human liver microsomes, respectively. Taken together, our results indicate that both CYP2C9 and CYP2C19 are involved in tolbutamide hydroxylation by human liver microsomes, and that CYP2C19 underlies at least 14 to 22% of tolbutamide metabolism. Although expression of CYP2C19 in human liver is less than that of CYP2C9, it may play an important role in tolbutamide disposition in subjects expressing either high levels of CYP2C19 or a catalytically deficient CYP2C9 enzyme. PMID- 10681383 TI - Human cytochrome P-450 3A4: in vitro drug-drug interaction patterns are substrate dependent. AB - Testosterone, terfenadine, midazolam, and nifedipine, four commonly used substrates for human cytochrome P-450 3A4 (CYP3A4), were studied in pairs in human liver microsomes and in microsomes from cells containing recombinant human CYP3A4 and P-450 reductase, to investigate in vitro substrate-substrate interaction with CYP3A4. The interaction patterns between compounds with CYP3A4 were found to be substrate-dependent. Mutual inhibition, partial inhibition, and activation were observed in the testosterone-terfenadine, testosterone-midazolam, or terfenadine-midazolam interactions. However, the most unusual result was the interaction between testosterone and nifedipine. Although nifedipine inhibited testosterone 6beta-hydroxylation in a concentration-dependent manner, testosterone did not inhibit nifedipine oxidation. Furthermore, the effect of testosterone and 7,8-benzoflavone on midazolam 1'-hydroxylation and 4 hydroxylation demonstrated different regiospecificities. These results may be explained by a model in which multiple substrates or ligands can bind concurrently to the active site of a single CYP3A4 molecule. However, the contribution of separate allosteric sites and conformational heterogeneity to the atypical kinetics of CYP3A4 can not be ruled out in this model. PMID- 10681384 TI - Butyrylcholinesterase accelerates cocaine metabolism: in vitro and in vivo effects in nonhuman primates and humans. AB - Butyrylcholinesterase (BChE) is known to metabolize cocaine in humans. In the present study, three different experiments were performed to determine whether the addition of horse serum-derived BChE would accelerate the metabolism of cocaine. In the first experiment, the addition of BChE to squirrel monkey plasma in vitro reduced the half-life of cocaine by over 80%, decreased the production of the metabolic product benzoylecgonine, and increased ecgonine methyl ester formation. The effect of BChE on cocaine metabolism was reversed by a specific BChE inhibitor. In the second, in vivo, experiment, exogenously administered BChE reduced peak cocaine concentrations when given to anesthetized squirrel monkeys. Finally, incubation of cocaine with added BChE in human plasma in vitro resulted in a decrease in cocaine half-life similar to that observed with squirrel monkey plasma. The magnitude of the decrease in cocaine half-life was proportional to the amount of added BChE. Together, these results indicate that exogenously administered BChE can accelerate cocaine metabolism in such a way as to potentially lessen the behavioral and toxic effects of cocaine. Therefore, BChE may be useful as a treatment for cocaine addiction and toxicity. PMID- 10681385 TI - Immunohistochemical localization of taurine in the male reproductive organs of the rat. AB - The amino acid taurine has been implicated in several aspects of reproductive system physiology. However, its localization in these organs has not been previously analyzed. The aim of this study was to characterize its distribution in male rat reproductive organs by immunohistochemical methods. Taurine was localized in the smooth muscle cells of the tissues studied and in the skeletal fibers of the cremaster muscle. In the testis, taurine was found in Leydig cells, vascular endothelial cells, and other interstitial cells. No immunoreactivity was observed in the cells of the seminiferous tubules, either in germ cells at all spermatogenic stages or in Sertoli cells. However, peritubular myoid cells were immunostained. Most epithelial cells of the efferent ducts were immunolabeled, whereas the epithelial cells of the rete testis (extratesticular segments), epididymis (caput, corpus, and cauda regions), and ductus deferens were unstained. However, most epithelial cells from the intratesticular segments of the rete were immunopositive. Some cells identified as intraepithelial macrophages and lymphocytes, apical cells, and narrow cells were intensely immunolabeled. Regional differences in the distribution of these cell types along the ducts studied were also noted. The possible functional roles for taurine in these cells are discussed. PMID- 10681386 TI - Heat-induced alterations in the localization of HSP72 and HSP73 as measured by indirect immunohistochemistry and immunogold electron microscopy. AB - The heat shock proteins are a family of stress-inducible proteins that act as molecular chaperones for nascent proteins and assist in protection and repair of proteins whose conformation is altered by stress. HSP72 and HSP73 are two major cytosolic/nuclear stress proteins of mammalian cells, with extensive sequence homology. HSP73 is constitutively expressed, whereas HSP72 is highly stress inducible. However, it is unclear why two isoforms are expressed and whether these two proteins have different functions in the cell. To assist in the delineation of function, we have completed a detailed study of the localization of HSP72 and HSP73 in the cell before and after heat stress, using two different methods of detection. By indirect immunohistochemistry, the localization of these two proteins is similar, cytoplasmic and nuclear in nonstressed cells with a translocation to nucleoli immediately after heat. By the more sensitive immunogold electron microscopy technique, differences in localization were noted. In nonstressed cells, HSP72 was primarily nuclear, localized in heterochromatic regions and in nucleoli. HSP73 was distributed throughout the cell, with most cytoplasmic label associated with mitochondria. Mitotic chromosomes were also heavily labeled. After stress, HSP72 concentrated in nuclei and nucleoli and HSP73 localized to nuclei, nucleoli, and cytoplasm, with increased label over mitochondria. These differences in localization suggest that the HSP72 and HSP73 may associate with different proteins or complexes and hence have different but overlapping functions in the cell. PMID- 10681387 TI - A qualitative and quantitative study on the enkephalinergic innervation of the pig gastrointestinal tract. AB - Enkephalins are involved in neural control of digestive functions such as motility, secretion, and absorption. To better understand their role in pigs, we analyzed the qualitative and quantitative distribution of enkephalin immunoreactivity (ENK-IR) in components of the intestinal wall from the esophagus to the anal sphincter. Immunohistochemical labelings were analyzed using conventional fluorescence and confocal microscopy. ENK-IR was compared with the synaptophysin immunoreactivity (SYN-IR). The results show that maximal ENK-IR levels in the entire digestive tract are reached in the myenteric plexuses and, to a lesser extent, in the external submucous plexus and the circular muscle layer. In the longitudinal muscle layer, ENK-IR was present in the esophagus, stomach, rectum, and anal sphincter, whereas it was absent from the duodenum to the distal colon. In the ENK-IR plexuses and muscle layers, more than 60% of the nerve fibers identified by SYN-IR expressed ENK-IR. No ENK-IR was observed in the internal submucous plexus and the mucosa; the latter was found to contain ENK-IR endocrine cells. These results strongly suggest that, in pigs, enkephalins play a major role in the regulatory mechanisms that underlie the neural control of digestive motility. PMID- 10681388 TI - cAMP-dependent Cl(-) channel protein (CFTR) and its mRNA are expressed in the secretory portion of human eccrine sweat gland. AB - Immunoreactive cystic fibrosis transport regulator (CFTR) proteins in human sweat ducts has been documented but CFTR expression in the secretory coil has remained uncertain. Using monoclonal antibodies (MAbs) against epitopes in the R-domain and C-terminus, we observed the following: Formalin fixation masks the CFTR epitopes but the epitopes are exposed by treatment with urea and heat (antigen retrieval). Pen-Fix fixation preserves CFTR epitopes. The secretory coil also expresses CFTR epitopes for the R-domain and C-terminus. An MAb against C terminus amino acids 1466-1480 coupled to keyhole limpet hemocyanin (MAb WC) stained dark cells predominantly. Staining by MAbs against the C-terminus was completely blocked by a C-terminus peptide. mRNA for CFTR was amplified by RT-PCR in both the duct and the secretory coil. In situ hybridization for CFTR mRNA after 3SR amplification indicates that mRNA is localized in the dark cells and perhaps also in the clear cell cytoplasm near the secretory coil. mRNA is present in both the luminal and basal duct cells. We conclude that CFTR is expressed equally well in both the duct and the secretory coil, suggesting that cAMP dependent Cl(-) channels are involved in regulation of sweat secretion and duct absorption. PMID- 10681389 TI - Effect of fixation and epitope retrieval on BrdU indices in mammary carcinomas. AB - Studies in which 5-bromo-2'-deoxyuridine (BrdU) is used to quantify rates of cell proliferation are conducted prospectively. Therefore, the opportunity exists to select conditions that optimize detection of the BrdU epitope. The objective of this study was to quantify the extent to which the BrdU epitope was masked by formalin vs methacarn fixation in the assessment of cell proliferation. Mammary carcinomas from animals pulse-labeled with BrdU were trisected. A portion was frozen and the remaining two portions were fixed in 10% neutral buffered formalin or methacarn for 24 hr, processed, embedded in paraffin, and sections stained for incorporated BrdU using a peroxidase immunohistochemical staining technique. Antigen retrieval techniques also were applied to formalin-fixed sections. Fixation in methacarn gave the highest labeling index (16.4%), which was comparable to that observed in unfixed frozen sections (17.5%). Formalin fixation alone dramatically suppressed the labeling index (0.3%), which was only partially recovered using various antigen retrieval techniques (2.1-8.1%). Methacarn fixation is recommended for prospective studies in which BrdU detection is planned because of the quantitative recovery of epitope and the simplicity of the approach. PMID- 10681390 TI - Integrin expression during epithelial migration and restratification in the tenascin-C-deficient mouse cornea. AB - In the unwounded cornea, tenascin-C localizes to a short stretch of the basement membrane zone at the corneoscleral junction or limbus. To determine whether the function of the limbus is affected by the absence of tenascin-C, mice possessing a deletion of tenascin-C and strain-matched wild-type mice are used in corneal debridement wounding experiments. The expression of integrins (alpha3, alpha9, and beta4) in the tenascin-C knockout corneas is evaluated by producing polyclonal cytoplasmic domain antipeptide sera and performing immunofluorescence microscopy. In addition, we evaluate the localization of several other proteins involved in wound healing, including fibronectin, laminin beta1, nidogen/entactin, and VCAM-1, in both the tenascin knockout and wild-type mice. There are no differences in healing rate, scarring, or neovascularization after corneal debridement wounds. alpha9 integrin is expressed at the limbal border of unwounded tenascin-C knockout animals and is upregulated during migration only after the larger wounds. At 8 weeks after larger wounds, the localization of alpha9 again becomes restricted to the limbal border. Results show that tenascin C is not required for development or maintenance of the corneal limbus or for normal re-epithelialization of corneal epithelial cells after debridement wounding. PMID- 10681391 TI - Immunohistochemical study of the expression of MUC6 mucin and co-expression of other secreted mucins (MUC5AC and MUC2) in human gastric carcinomas. AB - To investigate the expression of MUC6 mucin in gastric carcinomas, we generated a novel monoclonal antibody (MAb CLH5) using an MUC6 synthetic peptide. MAb CLH5 reacted exclusively with the MUC6 peptide and with native and deglycosylated mucin extracts from gastric tissues. MAb CLH5 immunoreactivity was observed in normal gastric mucosa restricted to pyloric glands of the antrum and mucopeptic cells of the neck zone of the body region. In a series of 104 gastric carcinomas, 31 (29.8%) were immunoreactive for MUC6. The expression of MUC6 was not associated with histomorphological type or with clinicopathological features of the carcinomas. Analysis of the co-expression of MUC6 with other secreted mucins (MUC5AC and MUC2) in 20 gastric carcinomas revealed that different mucin core proteins are co-expressed in 55% of the cases. MUC6 was co-expressed and co localized with MUC5AC in 45% and with MUC2 in 5% of the cases. Expression of MUC2 alone was observed in 25% of the cases. All carcinomas expressing MUC2 mucin in more than 50% of the cells were of the mucinous type according to the WHO classification. The co-expression of mucins was independent of the histomorphological type and stage of the tumors. In conclusion, we observed, using a novel well-characterized MAb, that MUC6 is a good marker of mucopeptic cell differentiation and is expressed in 30% of gastric carcinomas, independent of the clinicopathological features of the cases. Furthermore, we found that co expression and co-localization of mucins in gastric carcinomas is independent of histomorphology and staging. Finally, we observed that intestinal mucin MUC2 is expressed as the most prominent mucin of the mucins tested in mucinous-type gastric carcinomas. PMID- 10681392 TI - Expression of prolactin mRNA in rat mammary gland during pregnancy and lactation. AB - We studied the expression of prolactin (PRL) mRNA in the mammary gland of resting, pregnant, lactating, and weanling rats using in situ and solution reverse transcriptase-polymerase chain reaction (RT-PCR). In mid- to late pregnancy and throughout lactation, PRL mRNA was detected in both in situ and solution RT-PCR. These PRL mRNA signals were clearly identified in the cytoplasm of alveolar and ductal mammary epithelial cells by the in situ RT-PCR method. In mid- to late pregnancy, such as at the initiating point of PRL mRNA expression, we confirmed in some cases a lack of PRL mRNA by solution RT-PCR. In addition, in the early weaning phase, no signals were detected by solution RT-PCR. However, slight focal signals were detected in some poorly vacuolated cytoplasm of regressing acinar cells by in situ RT-PCR. These findings suggest that PRL mRNA in rat mammary gland begins in mid- to late pregnancy in parallel with the development of the mammary gland, continues throughout lactation, and declines in the early phase of weaning, with regression of mammary epithelial cells. PMID- 10681393 TI - The cadherin-catenin complex is expressed alternately with the adenomatous polyposis coli protein during rat incisor amelogenesis. AB - E-cadherin, a calcium-dependent cell-cell adhesion molecule, is expressed in highly specific spatiotemporal patterns throughout metazoan development, notably at sites of embryonic induction. E-cadherin also plays a critical role in regulating cell motility/adhesion, cell proliferation, and apoptosis. We have used the continuously erupting rat incisor as a system for examining the expression of E-cadherin and the associated catenins [alpha-, beta-, gamma catenin (plakoglobin) and p120(ctn)] during amelogenesis. Using immunhistochemical techniques, we observed expression of alpha-catenin and gamma catenin in ameloblasts throughout amelogenesis. In contrast, expression of E cadherin, beta-catenin, and p120(ctn) was strong in presecretory, transitional, and reduced stage ameloblasts (Stages I, III, and V) but was dramatically lower in secretory and maturation stage ameloblasts (Stages II and IV). This expression alternates with the expression pattern we previously reported for the adenomatous polyposis coli protein (APC), a tumor suppressor that competes with E-cadherin for binding to beta-catenin. We suggest that alternate expression of APC and the cadherin-catenin complex is critical for the alterations in cell-cell adhesion and other differentiated cellular characteristics, such as cytoskeletal alterations, that are required for the formation of enamel by ameloblasts. PMID- 10681394 TI - Acidic fibroblast growth factor is present in the enteric nervous system of the large intestine. AB - Acidic fibroblast growth factor (aFGF) is a heparin binding protein that displays pleiotropic activity. The purpose of this study was to document the presence of the translated aFGF product, its mRNA, and its location in the colon. mRNA was extracted from bovine large intestine and reverse transcribed to cDNA. Nested primer PCR was used to determine the presence of mRNA using primers homologous to the previously published bovine aFGF cDNA. Purification of translated aFGF was performed using an established HPLC protocol. Western blot analysis of the HPLC fractions was performed using two epitope-independent antibodies against aFGF. Immunohistochemistry employed these antibodies to determine the locus of aFGF expression. The nested-primer PCR product of predicted size was homologous to the published bovine aFGF mRNA sequence, as determined by DNA sequencing. Intestinal aFGF had a mass similar to bovine aFGF isolated from other tissues, and immunocrossreacted with two peptide-based, epitope-independent anti-aFGF antisera on Western blotting. Immunohistochemical analysis of large intestine using these two independent antisera localized aFGF within the myenteric plexus. These data demonstrate that aFGF is present within the myenteric plexus of the enteric nervous system. PMID- 10681395 TI - Expression of glutamine synthetase in macrophages. AB - We studied the expression of glutamine synthetase in liver macrophages (Kupffer cells, KCs) in situ and in culture. Glutamine synthetase was detectable at the mRNA and protein level in freshly isolated and short-term-cultured rat liver macrophages. Enzyme activity and protein content were about 9% of that in liver parenchymal cells. In contrast, glutamine synthetase mRNA levels in liver macrophages apparently exceeded those in parenchymal liver cells (PCs). By use of confocal laser scanning microscopy and specific macrophage markers, immunoreactive glutamine synthetase was localized to macrophages in normal rat liver and normal human liver in situ. All liver macrophages stained positive for glutamine synthetase. In addition, macrophages in rat pancreas contained immunoreactive glutamine synthetase, whereas glutamine synthetase was not detectable at the mRNA and protein level in blood monocytes and RAW 264.7 mouse macrophages. No significant amounts of glutamine synthetase were found in isolated rat liver sinusoidal endothelial cells (SECs). The data suggest a constitutive expression of glutamine synthetase not only, as previously believed, in perivenous liver parenchymal cells but also in resident liver macrophages. PMID- 10681396 TI - Immunolocalization of collagen types II and III in single fibrils of human articular cartilage. AB - Type II and III fibrillar collagens were localized by immunogold electron microscopy in resin sections of human femoral articular cartilage taken from the upper radial zone in specimens from patients with osteoarthritis. Tissue samples stabilized by high-pressure cryofixation were processed by freeze-substitution, either in acetone containing osmium or in methanol without chemical fixatives, before embedding in epoxy or Lowicryl resin, respectively. Ultrastructural preservation was superior with osmium-acetone, although it was not possible to localize collagens by this method. In contrast, in tissue prepared by low temperature methods without chemical fixation, collagens were successfully localized with mono- or polyclonal antibodies to the helical (Types II and III) and amino-propeptide (Type III procollagen) domains of the molecule. Dual localization using secondary antibodies labeled with 5- or 10-nm gold particles demonstrated the presence of Types II and III collagen associated within single periodic banded fibrils. Collagen fibrils in articular cartilage are understood to be heteropolymers mainly of Types II, IX, and XI collagen. Our observations provide further evidence for the complexity of these assemblies, with the potential for interactions between at least 11 distinct collagen types as well as several noncollagenous components of the extracellular matrix. PMID- 10681397 TI - Analysis of antiphotobleaching reagents for use with FluoroNanogold in correlative microscopy. AB - Correlative microscopy is an important approach for bridging the resolution gap between fluorescence and electron microscopy. We have employed FluoroNanogold (FNG) as the detection system in these types of studies. This immunoprobe consists of a gold cluster compound to which a fluorochrome-labeled antibody is covalently linked. In these preparations, the fluorescence signal from FNG is first recorded then the gold cluster compound is subjected to a silver enhancement reaction before examination by electron microscopy. Potential complications are those associated with photochemical reactions that occur during fluorescence microscopy. We have evaluated this and some anti-photobleaching agents (i.e., 1,4-diazabicyclo[2.2.2]octane [DABCO],p-phenylenediamine [PPD], and N-propyl gallate [NPG]) for their utility with FNG in correlative microscopy. When DABCO was employed, the gold signal from FNG was dramatically diminished but the fluorescence signal was unaffected. The gold signal of DABCO-treated samples decreased to approximately 30% of that of the other samples. On the other hand, PPD and NPG did not adversely affect the FNG labeling. We recommend that either PPD or NPG be used and that DABCO be avoided as an antiphotobleaching reagent for this technique. PMID- 10681398 TI - Direct eye visualization of Cy5 fluorescence for immunocytochemistry and in situ hybridization. AB - Cyanine 5.18 (or Cy5) is a fluorochrome emitting in the long-red/far-red range, usually regarded as unsuitable for direct observation by the human eye. We describe here the optimization of a direct visualization approach to Cy5 labeling, based on a standard fluorescence microscope with mercury light excitation and applicable to both immunocytochemistry and fluorescent in situ hybridization. Crucial factors were (a) an excitation path in the microscope not absorbing light in the orange-red range, up to 640 nm, (b) a 588-640-nm excitation filter range, distinctly below the excitation optimum for Cy5, (c) a 650-700-nm emission filter range, transmitting the low-wavelength portion of Cy5 emission, and (d) high-efficiency filter set components allowing a narrow gap between excitation and emission ranges without visible cross-talk of excitation light in the emission path. PMID- 10681399 TI - Cytochrome P450 and arachidonic acid bioactivation. Molecular and functional properties of the arachidonate monooxygenase. AB - The demonstration of in vivo arachidonic acid epoxidation and omega-hydroxylation established the cytochrome P450 epoxygenase and omega/omega-1 hydroxylase as formal metabolic pathways and as members of the arachidonate metabolic cascade. The characterization of the potent biological activities associated with several of the cytochrome P450-derived eicosanoids suggested new and important functional roles for these enzymes in cellular, organ, and body physiology, including the control of vascular reactivity and systemic blood pressures. Past and current advances in cytochrome P450 biochemistry and molecular biology facilitate the characterization of cytochrome P450 isoforms responsible for tissue/organ specific arachidonic acid epoxidation and omega/omega-1 hydroxylation, and thus, the analysis of cDNA and/or gene specific functional phenotypes. The combined application of physiological, biochemical, molecular, and genetic approaches is beginning to provide new insights into the physiological and/or pathophysiological significance of these enzymes, their endogenous substrates, and products. PMID- 10681400 TI - n-3 PUFA dietary supplementation inhibits proliferation and store-operated calcium influx in thymoma cells growing in Balb/c mice. AB - The antitumor effect of daily individual administration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (2 g/kg body weight) in Balb/c mice bearing a transplantable thymoma was investigated. Mice received oleic acid (control group), EPA and DHA ethyl esters starting 10 days before tumor inoculation. Analysis of phospholipid composition of neoplastic cell revealed that EPA and DHA levels were significantly increased (63 and 22% increase) after EPA and DHA treatments, respectively. Conversely, decreased levels of arachidonic acid were found in both cases (19 and 24% decrease in EPA and DHA groups, respectively). EPA and DHA delayed the appearance of macroscopic ascites (100% of animal, from 7 to 28 days), prolonged animal survival (100% of animal, from 22 to 32 and 33 days, respectively) and reduced the percentage of proliferating tumor cells detected by immunostaining of proliferation cell nuclear antigen (PCNA) (80 and 85% decrease, respectively). Moreover, the regulatory effects of these dietary n; 3 fatty acids on the influx of Ca(2+), activated by depletion of intracellular stores with thapsigargin (Tg), were investigated. By using a Ca(2+)-free/Ca(2+) reintroduction protocol and Fura-2 as fluorescent indicator of intracellular free Ca(2+)([Ca(2+)](i)), we observed that EPA and DHA treatments markedly decreased Tg-induced rise in [Ca(2+)](i) (49 and 37% decrease, respectively). This effect was related to the inhibition of the store-operated Ca(2+) influx, as confirmed also by Mn(2+) influx experiments. The inhibitory action of EPA and DHA on the store-operated Ca(2+) influx could explain, at least in part, their antitumoral activity, as this Ca(2+) mobilization pathway appears to be involved in the cell signaling occurring in non-excitable cells to evoke many cellular processes, including cell proliferation. PMID- 10681401 TI - Measurement of cholesterol gallstone growth in vitro. AB - Methods to study growth of gallstones in the laboratory have not been reported. We here present such a method. Human cholesterol gallstones were harvested from patients with multiple nearly identical stones. The gallstones were washed and added to supersaturated model biles and the formation of cholesterol crystals and the increases in mass of human cholesterol gallstones were studied concurrently, over a period of weeks, using nephelometry and a microbalance, respectively. All stones incubated in model biles supersaturated with cholesterol increased in mass. Increases in the degree of supersaturation of cholesterol in the model biles resulted in increased growth of stones. The mass increases, the growth rates, and the spatial orientation of accreted crystalline cholesterol differed among various stone types. The kinetics and structures of stone growth were similar when the stones were incubated in supersaturated, native, human gallbladder biles. The structure of accreted cholesterol was the same as found on the surface of some human gallstones that were harvested during apparent active growth in situ. This simple method allows accurate measurements of stone growth in vitro, in patterns that mimic stone growth in vivo, and is useful for studies on the relationships of gallstone growth and the kinetics of cholesterol crystallization. PMID- 10681402 TI - Plasma 24S-hydroxycholesterol (cerebrosterol) is increased in Alzheimer and vascular demented patients. AB - Alzheimer's disease (AD) is characterized by the presence of senile plaques, neurofibrillary tangles, and neuronal cell loss associated with membrane cholesterol release. 24S-hydroxycholesterol (24S-OH-Chol) is an enzymatically oxidized product of cholesterol mainly synthesized in the brain. We tested the hypothesis that plasma levels of this oxysterol could be used as a putative biochemical marker for an altered cholesterol homeostasis in the brain of AD patients. Thirty patients with clinical criteria for AD, 30 healthy volunteers, 18 depressed patients, and 12 patients with vascular dementia (non-Alzheimer demented) were studied. Plasma concentrations of 24S-OH-Chol were assayed by isotope dilution;-mass spectrometry, cholesterol was measured enzymatically, and apolipoprotein E (apoE) was genotyped by polymerase chain reaction and restricted fragment length polymorphism. The concentration of 24S-OH-Chol in AD and non Alzheimer demented patients was modestly but significantly higher than in healthy controls and in depressed patients. There was no significant difference in the concentrations of 24S-OH-Chol between depressed patients and healthy controls nor between AD and non-Alzheimer demented patients. The apoE straightepsilon4 allele influences plasma 24S-OH-Chol. However, this influence could be completely accounted for by the elevated plasma cholesterol in apoE4 hetero- or homozygotes. Plasma 24S-OH-Chol levels correlated negatively with the severity of dementia. AD and vascular demented patients appear to have higher circulating levels of 24S-OH Chol than depressed patients and healthy controls. We speculate that 24S-OH-Chol plasma levels may potentially be used as an early biochemical marker for an altered cholesterol homeostasis in the central nervous system. 24S hydroxycholesterol (cerebrosterol) is increased in Alzheimer and vascular demented patients. PMID- 10681403 TI - Mechanism of the stimulatory action of okadaic acid on lipolysis in rat fat cells. AB - Okadaic acid was found to induce concentration- and time-dependent lipolysis in rat fat cells in the absence of lipolytic hormones, but it did not significantly increase the total hormone-sensitive lipase (HSL) activity in these fat cells, the activity of HSL extracted from fat layer and that of HSL in the supernatant of homogenized fat cells. Western blotting of fat cell homogenate fractions with an antiserum raised against synthetic peptide derived from rat HSL showed that HSL protein shifted from the supernatant to the fat layer in response to okadaic acid, which increased the HSL protein content on the fat layer and concomitantly reduced that of the supernatant, concentration- and time-dependently. Sonication of the fat cells abolished their responsiveness to okadaic acid. The lipolytic action of okadaic acid was examined and its site was identified using a cell-free system comprising lipid droplets isolated from rat fat cells and HSL. Okadaic acid induced lipolysis in this cell-free system and sonication of the lipid droplets caused disappearance of lipolytic action of okadaic acid. Okadaic acid failed to stimulate lipolysis in a cell-free system comprising HSL and artificial lipid droplets (trioleoylglycerol emulsified with gum arabic) instead of lipid droplets isolated from rat fat cells. These results suggest that okadaic acid does not increase the catalytic activity of HSL but induces translocation of HSL to the lipid droplets isolated from rat fat cells. The site of the lipolytic action of okadaic acid in relation to the interaction between HSL and lipid droplet is discussed. PMID- 10681404 TI - Relative roles of LDLr and LRP in the metabolism of chylomicron remnants in genetically manipulated mice. AB - Remnant-like emulsions labeled with cholesteryl [(13)C]-oleate were prepared with lipid compositions similar to remnants derived from triacylglycerol-rich lipoproteins. When injected into the bloodstream of conscious mice, the remnant like emulsions were metabolized in the liver leading to the appearance of (13)CO(2) in the breath. Previously, using this technique, we found that remnant metabolism was significantly impaired but not completely inhibited in mice lacking low density lipoprotein receptors (LDLr). We have now found in mice with non-functional low density lipoprotein receptor-related protein (LRP) that breath enrichment of (13)CO(2) was significantly decreased, indicating that the LRP also plays an important role in the metabolism of chylomicron remnants (CR). The enrichment of (13)CO(2) in the expired breath was negligible in mice lacking both LDLr and receptor-associated protein (-/-), essential for normal function of LRP. In mice pre-injected with gluthatione S-transferase-receptor-associated protein to block LRP binding, there was a marked inhibition of the appearance of (13)CO(2) in the expired breath of homozygous LDLr-deficient mice, supporting the role of LRP in vivo. Whether or not LDLr were present, in mouse and human fibroblast cells human apoE3 or E4 but not apoE2 were essential for binding of remnant-like emulsions, while lactoferrin and suramin completely inhibited binding. We conclude that in normal mice LDLr are important for the physiological metabolism of CR. When LDLr are absent the evidence supports a role for the LRP in the uptake of CR in liver cells and in fibroblasts, with binding characteristics for CR-associated apoE similar to LDLr. PMID- 10681405 TI - Adrenal and liver in normal and cld/cld mice synthesize and secrete hepatic lipase, but the lipase is inactive in cld/cld mice. AB - Combined lipase deficiency (cld) is a recessive mutation in mice that causes a severe lack of lipoprotein lipase (LPL) and hepatic lipase (HL) activities, hyperlipemia, and death within 3 days after birth. Earlier studies showed that inactive LPL and HL were synthesized by cld/cld tissues and that LPL synthesized by cld/cld brown adipocytes was retained in their ER. We report here a study of HL in liver, adrenal, and plasma of normal newborn and cld/cld mice. Immunofluorescence studies showed HL was present in extracellular space, but not in cells, in liver and adrenal of both normal and cld/cld mice. When protein secretion was blocked with monensin, HL was retained intracellularly in liver cell cultures and in incubated adrenal tissues of both groups of mice. These findings demonstrated that HL was synthesized and secreted by liver and adrenal cells in normal newborn and cld/cld mice. HL activities in liver, adrenal, and plasma in cld/cld mice were very low, <8% of that in normal newborn mice, indicating that HL synthesized and secreted by cld/cld cells was inactive. Livers of both normal newborn and cld/cld mice synthesized LPL, but the level of LPL activity in cld/cld liver was very low, <9% of that in normal liver. Immunofluorescence studies showed that LPL was present intracellularly in liver of cld/cld mice, indicating that LPL was synthesized but not secreted by cld/cld liver cells. Immunofluorescent LPL was not found in normal newborn liver cells unless the cells were treated with monensin, thus demonstrating that normal liver cells synthesized and secreted LPL. Livers of both groups of mice contained an unidentified alkaline lipase activity which accounted for 34-54% of alkaline lipase activity in normal and 65% of that in cld/cld livers. Our findings indicate that liver and adrenal cells synthesized and secreted HL in both normal newborn and cld/cld mice, but the lipase was inactive in cld/cld mice. That cld/cld liver cells secreted inactive HL while retaining inactive LPL indicates that these closely related lipases were processed differently. PMID- 10681406 TI - Cholesterol and oxysterol metabolism and subcellular distribution in macrophage foam cells. Accumulation of oxidized esters in lysosomes. AB - Cholesterol- and cholesteryl ester-rich macrophage foam cells, characteristic of atherosclerotic lesions, are often generated in vitro using oxidized low density lipoprotein (OxLDL). However, relatively little is known of the nature and extent of sterol deposition in these cells or of its relationship to the foam cells formed in atherosclerotic lesions. The purpose of this study was to examine the content and cellular processing of sterols in OxLDL-loaded macrophages, and to compare this with macrophages loaded with acetylated LDL (AcLDL; cholesteryl ester-loaded cells containing no oxidized lipids) or 7-ketocholesterol-enriched acetylated LDL (7KCAcLDL; cholesteryl ester-loaded cells selectively supplemented with 7-ketocholesterol (7KC), the major oxysterol present in OxLDL). Both cholesterol and 7KC and their esters were measured in macrophages after uptake of these modified lipoproteins. Oxysterols comprised up to 50% of total sterol content of OxLDL-loaded cells. Unesterified 7KC and cholesterol partitioned into cell membranes, with no evidence of retention of either free sterol within lysosomes. The cells also contained cytosolic, ACAT-derived, cholesteryl and 7 ketocholesteryl esters. The proportion of free cholesterol and 7KC esterified by ACAT was 10-fold less in OxLDL-loaded cells than in AcLDL or 7KCAcLDL-loaded cells. This poor esterification rate in OxLDL-loaded cells was partly caused by fatty acid limitation. OxLDL-loaded macrophages also contained large (approximately 40-50% total cell sterol content) pools of oxidized esters, containing cholesterol or 7KC esterified to oxidized fatty acids. These were insensitive to ACAT inhibition, very stable and located in lysosomes, indicating resistance to lysosomal esterases. Macrophages loaded with OxLDL do not accumulate free sterols in their lysosomal compartment, but do accumulate lysosomal deposits of OxLDL-derived cholesterol and 7-ketocholesterol esterified to oxidized fatty acids. The presence of similar deposits in lesion foam cells would represent a pool of sterols that is particularly resistant to removal. PMID- 10681407 TI - PLTP activity in premenopausal women. Relationship with lipoprotein lipase, HDL, LDL, body fat, and insulin resistance. AB - Plasma phospholipid transfer protein (PLTP) is thought to play a major role in the facilitated transfer of phospholipids between lipoproteins and in the modulation of high density lipoprotein (HDL) particle size and composition. However, little has been reported concerning the relationships of PLTP with plasma lipoprotein parameters, lipolytic enzymes, body fat distribution, insulin, and glucose in normolipidemic individuals, particularly females. In the present study, 50 normolipidemic healthy premenopausal females were investigated. The relationships between the plasma PLTP activity and selected variables were assessed. PLTP activity was significantly and positively correlated with low density lipoprotein (LDL) cholesterol (r(s) = 0.53), apoB (r(s) = 0.44), glucose (r(s) = 0.40), HDL cholesterol (r(s) = 0.38), HDL(3) cholesterol (r(s) = 0.37), lipoprotein lipase activity (r(s) = 0.36), insulin (r(s) = 0.33), subcutaneous abdominal fat (r(s) = 0.36), intra-abdominal fat (r(s) = 0.29), and body mass index (r(s) = 0.29). HDL(2) cholesterol, triglyceride, and hepatic lipase were not significantly related to PLTP activity. As HDL(2) can be decreased by hepatic lipase and hepatic lipase is increased in obesity with increasing intra-abdominal fat, the participants were divided into sub-groups of non-obese (n = 35) and obese (n = 15) individuals and the correlation of PLTP with HDL(2) cholesterol was re-examined. In the non-obese subjects, HDL(2) cholesterol was found to be significantly and positively related to PLTP activity (r(s) = 0.44). Adjustment of the HDL(2) values for the effect of hepatic lipase activity resulted in a significant positive correlation between PLTP and HDL(2) (r(s) = 0.41), indicating that the strength of the relationship between PLTP activity and HDL(2) can be reduced by the opposing effect of hepatic lipase on HDL(2) concentrations. We conclude that PLTP-facilitated lipid transfer activity is related to HDL and LDL metabolism, as well as lipoprotein lipase activity, adiposity, and insulin resistance. PMID- 10681408 TI - Contribution of the hepatic lipase gene to the atherogenic lipoprotein phenotype in familial combined hyperlipidemia. AB - Familial combined hyperlipidemia (FCH) is a common genetic lipid disorder with a frequency of 1-2% in the population. In addition to the hypercholesterolemia and/or hypertriglyceridemia that affected individuals exhibit, small, dense LDL particles and decreased HDL-cholesterol levels are traits frequently associated with FCH. Recently, we reported that families with FCH and families enriched for coronary artery disease (CAD) share genetic determinants for the atherogenic lipoprotein phenotype (ALP), a profile presenting with small, dense LDL particles, decreased HDL-cholesterol levels, and increased triglyceride levels. Other studies in normolipidemic populations have shown that the hepatic lipase (HL) gene is linked to HDL-cholesterol levels and that a polymorphism within the HL promoter (-514C-->T) is associated with increased HDL-cholesterol levels as well as larger, more buoyant LDL particles. In the present study, we tested whether the HL gene locus also contributes to ALP in a series of Dutch FCH families using nonparametric sibpair linkage analysis and association analysis. Evidence for linkage of LDL particle size (P < 0.019), HDL-cholesterol (P < 0.003), and triglyceride levels (P < 0.026) to the HL gene locus was observed. A genome scan in a subset of these families exhibited evidence for linkage of PPD (LOD = 2.2) and HDL-cholesterol levels (LOD = 1.2) to the HL gene locus as well. The -514C-->T promoter polymorphism was significantly associated (P < 0.0001) with higher HDL-cholesterol levels in the unrelated males of this population, but not in unrelated females. No association was observed between the polymorphism and LDL particle size or triglyceride levels. Our results provide support that ALP is a multigenic trait and suggest that the relationship between small, dense LDL particles, HDL-cholesterol, and triglyceride levels in FCH families is due, in part, to common genetic factors. PMID- 10681409 TI - Markedly increased secretion of VLDL triglycerides induced by gene transfer of apolipoprotein E isoforms in apoE-deficient mice. AB - Apolipoprotein E (apoE) plays a key role in the receptor-mediated uptake of lipoproteins by the liver and therefore in regulating plasma levels of lipoproteins. ApoE may also facilitate hepatic secretion of very low density lipoprotein (VLDL) triglyceride (TG). We directly tested the hypothesis that reconstitution of hepatic apoE expression in adult apoE-deficient mice by gene transfer would acutely enhance VLDL-TG production and directly compared the three major human apoE isoforms using this approach. Second generation recombinant adenoviruses encoding the three major isoforms of human apoE (E2, E3, and E4) or a control virus were injected intravenously into apoE-deficient mice, resulting in acute expression of the apoE isoforms in the liver. Despite the expected decreases in total and VLDL cholesterol levels, apoE expression was associated with increased total and VLDL triglyceride levels (E2 > E4 > E3). The increase in TG levels significantly correlated with plasma apoE concentrations. In order to determine whether acute apoE expression influenced the rate of VLDL-TG production, additional experiments were performed. Three days after injection of adenoviruses, Triton WR1339 was injected to block lipolysis of TG-rich lipoproteins and VLDL-TG production rates were determined. Mice injected with control adenovirus had a mean VLDL-TG production rate of 74 +/- 7 micromol/h/kg. In contrast, VLDL-TG production rates in apoE-expressing mice were 363 +/- 162 micromol/h/kg, 286 +/- 175 micromol/h/kg, and 300 +/- 84 micromol/h/kg for apoE2, apoE3, and apoE4, respectively. The VLDL-TG production rates in apoE-expressing mice were all significantly greater than in control mice but were not significantly different from each other. In summary, acute expression of all three human apoE isoforms in livers of apoE-deficient mice markedly increased VLDL-TG production to a similar degree, consistent with the concept that apoE plays an important role in facilitating hepatic VLDL-TG production in an isoform independent manner. PMID- 10681410 TI - Binding of hepatic lipase to heparin. Identification of specific heparin-binding residues in two distinct positive charge clusters. AB - The interaction of hepatic lipase (HL) with heparan sulfate is critical to the function of this enzyme. The primary amino acid sequence of HL was compared to that of lipoprotein lipase (LPL), a related enzyme that possesses several putative heparin-binding domains. Of the three putative heparin-binding clusters of LPL (J. Biol. Chem. 1994. 269: 4626-4633; J. Lipid Res. 1998. 39: 1310-1315), one was conserved in HL (Cluster 1; residues Lys 297-Arg 300 in rat HL) and two were partially conserved (Cluster 2; residues Asp 307-Phe 320, and Cluster 4; residues Lys 337, and Thr 432-Arg 443). Mutants of HL were generated in which potential heparin-binding residues within Clusters 1 and 4 were changed to Asn. Two chimeras in which the LPL heparin-binding sequences of Clusters 2 and 4 were substituted for the analogous HL sequences were also constructed. These mutants were expressed in Chinese hamster ovary (CHO) cells and assayed for heparin binding ability using heparin-Sepharose chromatography and a CHO cell-binding assay. The results suggest that residues within the homologous Cluster 1 region (Lys 297, Lys 298, and Arg 300), as well as some residues in the partially conserved Cluster 4 region (Lys 337, Lys 436, and Arg 443), are involved in the heparin binding of hepatic lipase. In the cell-binding assay, heparan sulfate binding affinity equal to that of LPL was seen for the RHL chimera mutant that possessed the Cluster 4 sequence of LPL. Mutation of Cluster 1 residues of HL resulted in a major reduction in heparin binding ability as seen in both the cell binding assay and the heparin-Sepharose elution profile. These results suggest that Cluster 1, the N-terminal heparin-binding domain, is of primary significance in RHL. This is different for LPL: mutations in the C-terminal binding domain (Cluster 4) cause a more significant shift in the salt required for elution from heparin-Sepharose than mutations in the N-terminal domain (Cluster 1). PMID- 10681411 TI - Phospholipid transfer protein gene knock-out mice have low high density lipoprotein levels, due to hypercatabolism, and accumulate apoA-IV-rich lamellar lipoproteins. AB - Phospholipid transfer protein gene knock-out (Pltp KO) mice have defective transfer of very low density lipoprotein (VLDL) phospholipids into high density lipoprotein (HDL) and markedly decreased HDL levels (Jiang et al. 1999. J. Clin. Invest. 103: 907-914). These animals also accumulated VLDL- and LDL-sized lipoproteins on a high saturated fat diet. The goals of this study were to further characterize the abnormal lipoproteins of Pltp KO mice and to determine the mechanisms responsible for low HDL levels. A lipoprotein fraction enriched in lamellar structures was isolated from the low density lipoprotein (LDL) region and was shown to be phospholipid- and free cholesterol-rich and to have apoA-IV (55%) and apoE (25%) as major apolipoproteins. The lamellar lipoproteins accumulating in these mice probably represent surface material derived from triglyceride-rich lipoproteins (TRL). The HDL was found to be protein-rich (primarily apoA-I) and specifically depleted in phosphatidylcholine (PC) (28% in wild-type mice (WT) vs. 15% in Pltp KO mice, P < 0.001). Unexpectedly, turnover studies using autologous HDL revealed a profound 4-fold increase in the catabolism of HDL protein and cholesteryl ester in Pltp KO mice compared to wild type, with minor differences in synthesis rates. In contrast, injection of WT mouse HDL into Pltp KO mice showed only a 2-fold increase in fractional catabolism. Reminiscent of the defect in Tangier disease, the failure of transfer of PC from TRL into the HDL fraction results in dramatic hypercatabolism of HDL. These results suggest that defective phospholipid transfer from TRL into HDL, arising from decreased lipolysis or decreased PLTP activity, could lead to hypoalphalipoproteinemia characterized by hypercatabolism of HDL protein. lipoprotein levels, due to hypercatabolism, and accumulate apoA-IV-rich lamellar lipoproteins. PMID- 10681412 TI - Regulation of calcium signalling by docosahexaenoic acid in human T-cells. Implication of CRAC channels. AB - We elucidated the role of docosahexaenoic acid (DHA) on the increases in free intracellular calcium concentrations, [Ca(2+)]i, in human (Jurkat) T-cell lines. DHA evoked an increase in [Ca(2+)]i in a dose-dependent manner in these cells. Anti-CD3 antibody, known to stimulate increases in Ca(2+) from endoplasmic reticulum (ER) via the production of inositol trisphosphate, also evoked increases in [Ca(2+)]i in Jurkat T-cells. We also used thapsigargin which inhibits Ca(2+)-ATPase of the ER and, therefore, increases Ca(2+) in the cytosol. Interestingly, addition of DHA during the thapsigargin-induced peak response exerted an additive effect on the increases in [Ca(2+)]i in human T-cells, indicating that the mechanisms of action of these two agents are different. However, the DHA-induced calcium response was not observed when this agent was added during the anti-CD3-induced calcium peak, though its addition resulted in a prolonged and sustained calcium response as a function of time, suggesting that DHA recruits calcium, in part, from the ER pool and the prolonged response may be due to Ca(2+) influx. In the medium containing 0% Ca(2+), the DHA-evoked response on the increases in [Ca(2+)]i was significantly curtailed as compared to that in 100% Ca(2+) medium, supporting the notion that the response of the DHA is also due, in part, to the opening of calcium channels. Furthermore, preincubation of cells with tyrphostin A9, an inhibitor of Ca(2+) release-activated Ca(2+) (CRAC) channels also significantly curtailed the DHA-induced sustained response on the increases in [Ca(2+)]i in these cells. These results suggest that DHA induces an increase in [Ca(2+)]i via the ER pool and the opening of CRAC channels in human T cells. PMID- 10681413 TI - Lipolytic remnants of human VLDL produced in vitro. Effect of HDL levels in the lipolysis mixtures on the apoCs to apoE ratio and metabolic properties of VLDL core remnants. AB - To determine the role of high-density lipoprotein (HDL) as an acceptor of lipolytic surface remnants of very low density lipoprotein (VLDL) in the metabolism of VLDL core remnants, we examined the effect of HDL levels in the VLDL lipolysis mixture on 1) the morphology and the apoCs to E ratio in VLDL core remnants and 2) the metabolic properties of VLDL core remnants in human hepatoma cell line HepG2 and human hepatocytes in the primary culture. Normolipidemic VLDL was lipolyzed in vitro by purified bovine milk lipoprotein lipase (LpL) in a lipolysis mixture containing a physiologic level of VLDL and albumin (30 mg VLDL cholesterol (CH)/dl and 6% albumin) in the absence and presence of either a low HDL level (VLDL-CH:HDL-CH = 3:1) or a high HDL level (VLDL-CH:HDL-CH = 1:4). Lipolysis of VLDL in either the absence or presence of HDL resulted in the hydrolysis of >85% of VLDL-triglycerides (TG) and the conversion of VLDL into smaller and denser particles. In the absence of HDL, heterogeneous spherical particles with numerous surface vesicular materials were produced. In the presence of low or high HDL, spherical particles containing some or no detectable vesicular surface components were produced. The apoCs to apoE ratios, as determined by densitometric scanning of the SDS polyacrylamide gradient gel, were 2.89 in control VLDL and 2.27, 0.91, and 0.22 in VLDL core remnants produced in the absence and in the presence of low and high HDL levels, respectively. In vitro lipolysis of VLDL markedly increased binding to HepG2 cells at 4 degrees C and internalization and degradation by human hepatocytes in primary culture at 37 degrees C. However, the HDL-mediated decrease in the apoCs to apoE ratio had a minimal effect on binding, internalization, and degradation of VLDL core remnants by HepG2 cells and human hepatocytes in primary culture. In order to determine whether HepG2 bound VLDL and VLDL core remnants are deficient in apoCs, (125)I labeled VLDL and VLDL core remnants were added to HepG2 culture medium at 4 degrees C. The bound particles were released by heparin, and the levels of (125)I labeled apoCs and apoE, relative to apoB, in the released particles were examined. When compared with those initially added to culture medium, the VLDL and VLDL core remnants released from HepG2 cells had a markedly increased (113%) level of apoE and a reduced (30-39%), but not absent, level of apoCs. We conclude that apoCs, as a minimum structural and/or functional component of VLDL and VLDL core remnants, may not have an inhibitory effect on the binding of VLDL or VLDL core remnants to hepatic apoE receptors. PMID- 10681414 TI - Ileal bile acid transport regulates bile acid pool, synthesis, and plasma cholesterol levels differently in cholesterol-fed rats and rabbits. AB - We investigated the effect of ileal bile acid transport on the regulation of classic and alternative bile acid synthesis in cholesterol-fed rats and rabbits. Bile acid pool sizes, fecal bile acid outputs (synthesis rates), and the activities of cholesterol 7alpha-hydroxylase (classic bile acid synthesis) and cholesterol 27-hydroxylase (alternative bile acid synthesis) were related to ileal bile acid transporter expression (ileal apical sodium-dependent bile acid transporter, ASBT). Plasma cholesterol levels rose 2.1-times in rats (98 +/- 19 mg/dl) and 31-times (986 +/- 188 mg/dl) in rabbits. The bile acid pool size remained constant (55 +/- 17 mg vs. 61 +/- 18 mg) in rats but doubled (254 +/- 46 to 533 +/- 53 mg) in rabbits. ASBT protein expression did not change in rats but rose 31% (P < 0.05) in rabbits. Fecal bile acid outputs that reflected bile acid synthesis increased 2- and 2.4-times (P < 0.05) in cholesterol-fed rats and rabbits, respectively. Cholesterol 7alpha-hydroxylase activity rose 33% (24 +/- 2.4 vs. 18 +/- 1.6 pmol/mg/min, P < 0.01) and mRNA levels increased 50% (P < 0.01) in rats but decreased 68% and 79%, respectively, in cholesterol-fed rabbits. Cholesterol 27-hydroxylase activity remained unchanged in rats but rose 62% (P < 0.05) in rabbits. Classic bile acid synthesis (cholesterol 7alpha hydroxylase) was inhibited in rabbits because an enlarged bile acid pool developed from enhanced ileal bile acid transport. In contrast, in rats, cholesterol 7alpha-hydroxylase was stimulated but the bile acid pool did not enlarge because ASBT did not change. Therefore, although bile acid synthesis was increased via different pathways in rats and rabbits, enhanced ileal bile acid transport was critical for enlarging the bile acid pool size that exerted feedback regulation on cholesterol 7alpha-hydroxylase in rabbits. PMID- 10681415 TI - Apolipoprotein B metabolism and the distribution of VLDL and LDL subfractions. AB - Apolipoprotein B (apoB) metabolism was investigated in 20 men with plasma triglyceride 0.66-2.40 mmol/l and plasma cholesterol 3.95-6. 95 mmol/l. Kinetics of VLDL(1) (S(f) 60-400), VLDL(2) (S(f) 20-60), IDL (S(f) 12-20), and LDL (S(f) 0;-12) apoB were analyzed using a trideuterated leucine tracer and a multicompartmental model which allowed input into each fraction. VLDL(1) apoB production varied widely (from 5.4 to 26.6 mg/kg/d) as did VLDL(2) apoB production (from 0.18 to 8.4 mg/kg/d) but the two were not correlated. IDL plus LDL apoB direct production accounted for up to half of total apoB production and was inversely related to plasma triglyceride (r = -0.54, P = 0.009). Percent of direct apoB production into the IDL/LDL density range (r = 0.50, P < 0.02) was positively related to the LDL apoB fractional catabolic rate (FCR). Plasma triglyceride in these subjects was determined principally by VLDL(1) and VLDL(2) apoB fractional transfer rates (FTR), i.e., lipolysis. IDL apoB concentration was regulated mainly by the IDL to LDL FTR (r = -0.71, P < 0.0001). LDL apoB concentration correlated with VLDL(2) apoB production (r = 0.48, P = 0.018) and the LDL FCR (r = -0.77, P < 0. 001) but not with VLDL(1), IDL, or LDL apoB production. Subjects with predominantly small, dense LDL (pattern B) had lower VLDL(1) and VLDL(2) apoB FTRs, higher VLDL(2) apoB production, and a lower LDL apoB FCR than those with large LDL (pattern A). Thus, the metabolic conditions that favored appearance of small, dense LDL were diminished lipolysis of VLDL, resulting in a raised plasma triglyceride above the putative threshold of 1.5 mmol/l, and a prolonged residence time for LDL. This latter condition presumably permitted sufficient time for the processes of lipid exchange and lipolysis to generate small LDL particles. PMID- 10681417 TI - Anatomy of a proficient enzyme: the structure of orotidine 5'-monophosphate decarboxylase in the presence and absence of a potential transition state analog. AB - Orotidine 5'-phosphate decarboxylase produces the largest rate enhancement that has been reported for any enzyme. The crystal structure of the recombinant Saccharomyces cerevisiae enzyme has been determined in the absence and presence of the proposed transition state analog 6-hydroxyuridine 5'-phosphate, at a resolution of 2.1 A and 2.4 A, respectively. Orotidine 5'-phosphate decarboxylase folds as a TIM-barrel with the ligand binding site near the open end of the barrel. The binding of 6-hydroxyuridine 5'-phosphate is accompanied by protein loop movements that envelop the ligand almost completely, forming numerous favorable interactions with the phosphoryl group, the ribofuranosyl group, and the pyrimidine ring. Lysine-93 appears to be anchored in such a way as to optimize electrostatic interactions with developing negative charge at C-6 of the pyrimidine ring, and to donate the proton that replaces the carboxylate group at C-6 of the product. In addition, H-bonds from the active site to O-2 and O-4 help to delocalize negative charge in the transition state. Interactions between the enzyme and the phosphoribosyl group anchor the pyrimidine within the active site, helping to explain the phosphoribosyl group's remarkably large contribution to catalysis despite its distance from the site of decarboxylation. PMID- 10681418 TI - Induction of a bystander mutagenic effect of alpha particles in mammalian cells. AB - Ever since the discovery of X-rays was made by Rontgen more than a hundred years ago, it has always been accepted that the deleterious effects of ionizing radiation such as mutation and carcinogenesis are attributable mainly to direct damage to DNA. Although evidence based on microdosimetric estimation in support of a bystander effect appears to be consistent, direct proof of such extranuclear/extracellular effects are limited. Using a precision charged particle microbeam, we show here that irradiation of 20% of randomly selected A(L) cells with 20 alpha particles each results in a mutant fraction that is 3 fold higher than expected, assuming no bystander modulation effect. Furthermore, analysis by multiplex PCR shows that the types of mutants induced are significantly different from those of spontaneous origin. Pretreatment of cells with the radical scavenger DMSO had no effect on the mutagenic incidence. In contrast, cells pretreated with a 40 microM dose of lindane, which inhibits cell cell communication, significantly decreased the mutant yield. The doses of DMSO and lindane used in these experiments are nontoxic and nonmutagenic. We further examined the mutagenic yield when 5-10% of randomly selected cells were irradiated with 20 alpha particles each. Results showed, likewise, a higher mutant yield than expected assuming no bystander effects. Our studies provide clear evidence that irradiated cells can induce a bystander mutagenic response in neighboring cells not directly traversed by alpha particles and that cell-cell communication process play a critical role in mediating the bystander phenomenon. PMID- 10681419 TI - Secretory responses of intact glomus cells in thin slices of rat carotid body to hypoxia and tetraethylammonium. AB - We have developed a thin-slice preparation of whole rat carotid body that allows us to perform patch-clamp recording of membrane ionic currents and to monitor catecholamine secretion by amperometry in single glomus cells under direct visual control. In normoxic conditions (P(O(2)) approximately 140 mmHg; 1 mmHg = 133 Pa), most glomus cells did not have measurable secretory activity, but exposure to hypoxia (P(O(2)) approximately 20 mmHg) elicited the appearance of a large number of spike-like exocytotic events. This neurosecretory response to hypoxia was fully reversible and required extracellular Ca(2+) influx. The average charge of single quantal events was 46 +/- 25 fC (n = 218), which yields an estimate of approximately 140,000 catecholamine molecules per vesicle. Addition of tetraethylammonium (TEA; 2-5 mM) to the extracellular solution induced in most (>95%) cells tested (n = 32) a secretory response similar to that elicited by low P(O(2)). Cells nonresponsive to hypoxia but activated by exposure to high external K(+) were also stimulated by TEA. A secretory response similar to the responses to hypoxia and TEA was also observed after treatment of the cells with iberiotoxin to block selectively Ca(2+)- and voltage-activated maxi-K(+) channels. Our data further show that membrane ion channels are critically involved in sensory transduction in the carotid body. We also show that in intact glomus cells inhibition of voltage-dependent K(+) channels can contribute to initiation of the secretory response to low P(O(2)). PMID- 10681420 TI - A library of bacteriophage-displayed antibody fragments directed against proteins of the inner ear. AB - Bacteriophage display of antibodies provides a method for the generation of immunological reagents against rare and uncharacterized antigens. To ascertain the usefulness of this approach for the characterization of inner-ear proteins, we produced a bacteriophage-displayed antibody-fragment library directed against proteins from the bullfrog's sacculus. This library was probed for bacteriophage that bound to proteins present in a lysate of hair cells, the sensory receptors of the inner ear. The predominant bacteriophage clone after selection expressed an antibody fragment that recognized a single protein in the inner ear. This antigen occurred in both the nonsensory and sensory epithelia of the sacculus. The specificity of the antibody fragment indicates that our bacteriophage displayed library provides a useful source of immunological tools that should facilitate the identification and biochemical characterization of novel proteins in the inner ear. PMID- 10681421 TI - Localized neuronal activation in the zebra finch brain is related to the strength of song learning. AB - Songbirds (Oscines) learn their songs from a tutor. It is not known where in the brain the memories of these learned sounds are stored. Recent evidence suggests that song perception in songbirds involves neuronal activation in brain regions that have not traditionally been implicated in the control of song production or song learning, notably the caudal part of the neostriatum (NCM) and of the hyperstriatum ventrale. Zebra finch males (Taeniopygia guttata castanotis) were reared without their father and exposed to a tape-recorded song during the sensitive period for song learning. When, as adults, they were reexposed to the tutor song, the males showed increased expression of the protein products of the immediate early genes egr-1 (ZENK) and c-fos in the NCM and caudal hyperstriatum ventrale, but not in the conventional "song-control nuclei." The strength of the immediate early gene response (which is a reflection of neuronal activation) in the NCM correlated significantly and positively with the number of song elements that the birds had copied from the tutor song. These results show localized neural activation in response to tutor song exposure that correlates with the strength of song learning. PMID- 10681422 TI - The mouse Nkx-1.2 homeobox gene: alternative RNA splicing at canonical and noncanonical splice sites. AB - A mouse homeobox gene, Nkx-1.2, (previously termed Sax-1) that is closely related to the Drosophila NK-1/S59 gene was cloned, and genomic DNA and cDNA were sequenced. Nine Nkx-1.2 cDNA clones were found that correspond to three species of Nkx-1.2 mRNA that are formed by alternative splicing at conventional 5' donor and 3' acceptor splice sites; however, seven cDNA clones were found that correspond to three species of Nkx-1.2 mRNA from testes that have novel TG/AC 5' and 3' splice sites. The consensus splice sequences are: 5' donor, CC downward arrowTGGAAG; 3' acceptor, ACTTAC downward arrow. Predicted amino acid sequences suggest that some transcripts may be translated into proteins that lack part or all of the homeodomain. At least three bands of Nkx-1.2 mRNA were found in RNA from the testes. Nkx-1.2 mRNA was shown to be present in postmeiotic germ cells of the testis and in mature spermatozoa. Nkx-1.2 mRNA also was found in regions of the adult cerebral cortex, hippocampus, diencephalon, pons/medulla, and cerebellum. Nkx-1.2 mRNA was found in embryos in highest abundance in 10-day embryos; the mRNA levels decrease during further development. Nkx-1.2 mRNA also was found in discrete zones of the embryonic mesencephalon and myelencephalon. PMID- 10681423 TI - Elevated matrix metalloprotease and angiostatin levels in integrin alpha 1 knockout mice cause reduced tumor vascularization. AB - Integrin alpha1beta1 is a collagen receptor abundantly expressed on microvascular endothelial cells. As well as being the only collagen receptor able to activate the Ras/Shc/mitogen-activated protein kinase pathway promoting fibroblast cell proliferation, it also acts to inhibit collagen and metalloproteinase (MMP) synthesis. We have observed that in integrin alpha1-null mice synthesis of MMP7 and MMP9 was markedly increased compared with that of their wild-type counterparts. As MMP7 and MMP9 have been shown to generate angiostatin from circulating plasminogen, and angiostatin acts as a potent inhibitor of endothelial cell proliferation, we determined whether tumor vascularization was altered in the alpha1-null mice. Tumors implanted into alpha1-null mice showed markedly decreased vascularization, with a reduction in capillary number and size, which was accompanied by an increase in plasma levels of angiostatin due to the action of MMP7 and MMP9 on circulating plasminogen. In vitro analysis of alpha1-null endothelial cells revealed a marked reduction of their proliferation on both integrin alpha1-dependent (collagenous) and independent (noncollagenous) substrata. This reduction was prevented by culturing alpha1-null cells with plasma derived from plasminogen-null animals, thus omitting the source from which to generate angiostatin. Plasma from tumor-bearing alpha1-null animals uniquely inhibited endothelial cell growth, and this inhibition was relieved by the coaddition of either MMP inhibitors, or antibody to angiostatin. Integrin alpha1 deficient mice thus provide a genetically characterized model for enhanced angiostatin production and serve to reveal an unwanted potential side effect of MMP inhibition, increased tumor angiogenesis. PMID- 10681424 TI - Pw1/Peg3 is a potential cell death mediator and cooperates with Siah1a in p53 mediated apoptosis. AB - Induction of wild-type p53 in mouse fibroblasts causes cell cycle arrest at the G(1) phase, whereas coexpression of p53 and the protooncogene c-myc induces apoptosis. Although p53 transcriptional activity generally is required for both pathways, the molecular components mediating p53-dependent apoptosis are not well understood. To identify factors that could mediate p53-induced cell death, we used a comparative RNA differential display procedure. We have identified Pw1/Peg3 as a gene product induced during p53/c-myc-mediated apoptosis. Pw1/Peg3 is not induced during p53-mediated G(1) growth arrest nor by c-myc alone. Although it is not clear whether the induction of Pw1/Peg3 depends on p53 activity, we show that Pw1/Peg3 interacts with a p53-inducible gene product Siah1a. We demonstrate that coexpression of Pw1/Peg3 with Siah1a induces apoptosis independently of p53 whereas expression of Pw1/Peg3 or Siah1a separately has no effect on cell death. These data suggest that Siah1a and Pw1/Peg3 cooperate in the p53-mediated cell death pathway. Furthermore, we show that inhibiting Pw1/Peg3 activity blocks p53-induced apoptosis. The observation that Pw1/Peg3 is necessary for the p53 apoptotic response suggests a pivotal role for this gene in determining cell death versus survival. PMID- 10681425 TI - Induced defenses in response to an invading crab predator: an explanation of historical and geographic phenotypic change. AB - The expression of defensive morphologies in prey often is correlated with predator abundance or diversity over a range of temporal and spatial scales. These patterns are assumed to reflect natural selection via differential predation on genetically determined, fixed phenotypes. Phenotypic variation, however, also can reflect within-generation developmental responses to environmental cues (phenotypic plasticity). For example, water-borne effluents from predators can induce the production of defensive morphologies in many prey taxa. This phenomenon, however, has been examined only on narrow scales. Here, we demonstrate adaptive phenotypic plasticity in prey from geographically separated populations that were reared in the presence of an introduced predator. Marine snails exposed to predatory crab effluent in the field increased shell thickness rapidly compared with controls. Induced changes were comparable to (i) historical transitions in thickness previously attributed to selection by the invading predator and (ii) present-day clinal variation predicted from water temperature differences. Thus, predator-induced phenotypic plasticity may explain broad-scale geographic and temporal phenotypic variation. If inducible defenses are heritable, then selection on the reaction norm may influence coevolution between predator and prey. Trade-offs may explain why inducible rather than constitutive defenses have evolved in several gastropod species. PMID- 10681426 TI - The role of cavities in protein dynamics: crystal structure of a photolytic intermediate of a mutant myoglobin. AB - We determined the structure of the photolytic intermediate of a sperm whale myoglobin (Mb) mutant called Mb-YQR [Leu-(B10)-->Tyr; His(E7)-->Gln; Thr(E10)- >Arg] to 1.4-A resolution by ultra-low temperature (20 K) x-ray diffraction. Starting with the CO complex, illumination leads to photolysis of the Fe-CO bond, and migration of the photolyzed carbon monoxide (CO*) to a niche in the protein 8.1 A from the heme iron; this cavity corresponds to that hosting an atom of Xe when the crystal is equilibrated with xenon gas at 7 atmospheres [Tilton, R. F., Jr., Kuntz, I. D. & Petsko, G. A. (1984) Biochemistry 23, 2849-2857]. The site occupied by CO* corresponds to that predicted by molecular dynamics simulations previously carried out to account for the NO geminate rebinding of Mb-YQR observed in laser photolysis experiments at room temperature. This secondary docking site differs from the primary docking site identified by previous crystallographic studies on the photolyzed intermediate of wild-type sperm whale Mb performed at cryogenic temperatures [Teng et al. (1994) Nat. Struct. Biol. 1, 701-705] and room temperature [Srajer et al. (1996) Science 274, 1726-1729]. Our experiment shows that the pathway of a small molecule in its trajectory through a protein may be modified by site-directed mutagenesis, and that migration within the protein matrix to the active site involves a limited number of pre-existing cavities identified in the interior space of the protein. PMID- 10681427 TI - The bst locus on mouse chromosome 16 is associated with age-related subretinal neovascularization. AB - Ocular neovascularization is the leading cause of blindness in developed countries and often causes rapid loss of vision in age-related macular degeneration. Acute visual loss is most often due to hemorrhage from new vessels that have extended from the choroid into the subretinal space. Growth of abnormal vessels beneath the retina in this condition is known as subretinal neovascularization (SRN). Age-related animal models of macular degeneration and SRN have not been described. Current animal models of SRN depend on chemical or physical stimuli to initiate growth of subretinal vessels. The genes responsible for age-related human macular degeneration with SRN have not been firmly identified. We report an angiogenic phenotype in Bst/+ mice that is age-related, clinically evident, and resembles human SRN. This represents a spontaneous, genetically determined model of SRN. Bst/+ mice offer the possibility of exploring the molecular mechanisms of SRN without the need for exogenous agents. PMID- 10681428 TI - TOGA: an automated parsing technology for analyzing expression of nearly all genes. AB - We have developed an automated, high-throughput, systematic cDNA display method called TOGA, an acronym for total gene expression analysis. TOGA utilizes 8-nt sequences, comprised of a 4-nt restriction endonuclease cleavage site and adjacent 4-nt parsing sequences, and their distances from the 3' ends of mRNA molecules to give each mRNA species in an organism a single identity. The parsing sequences are used as parts of primer-binding sites in 256 PCR-based assays performed robotically on tissue extracts to determine simultaneously the presence and relative concentration of nearly every mRNA in the extracts, regardless of whether the mRNA has been discovered previously. Visualization of the electrophoretically separated fluorescent assay products from different extracts displayed via a Netscape browser-based graphical user interface allows the status of each mRNA to be compared among samples and its identity to be matched with sequences of known mRNAs compiled in databases. PMID- 10681429 TI - Crystal structure of the zymogen form of the group A Streptococcus virulence factor SpeB: an integrin-binding cysteine protease. AB - Pathogenic bacteria secrete protein toxins that weaken or disable their host, and thereby act as virulence factors. We have determined the crystal structure of streptococcal pyrogenic exotoxin B (SpeB), a cysteine protease that is a major virulence factor of the human pathogen Streptococcus pyogenes and participates in invasive disease episodes, including necrotizing fasciitis. The structure, determined for the 40-kDa precursor form of SpeB at 1.6-A resolution, reveals that the protein is a distant homologue of the papain superfamily that includes the mammalian cathepsins B, K, L, and S. Despite negligible sequence identity, the protease portion has the canonical papain fold, albeit with major loop insertions and deletions. The catalytic site differs from most other cysteine proteases in that it lacks the Asn residue of the Cys-His-Asn triad. The prosegment has a unique fold and inactivation mechanism that involves displacement of the catalytically essential His residue by a loop inserted into the active site. The structure also reveals the surface location of an integrin binding Arg-Gly-Asp (RGD) motif that is a feature unique to SpeB among cysteine proteases and is linked to the pathogenesis of the most invasive strains of S. pyogenes. PMID- 10681430 TI - Notch signaling and the determination of appendage identity. AB - The Notch signaling pathway defines an evolutionarily conserved cell-cell interaction mechanism that throughout development controls the ability of precursor cells to respond to developmental signals. Here we show that Notch signaling regulates the expression of the master control genes eyeless, vestigial, and Distal-less, which in combination with homeotic genes induce the formation of eyes, wings, antennae, and legs. Therefore, Notch is involved in a common regulatory pathway for the determination of the various Drosophila appendages. PMID- 10681431 TI - The xeroderma pigmentosum group C gene leads to selective repair of cyclobutane pyrimidine dimers rather than 6-4 photoproducts. AB - We investigated the contribution of the xeroderma pigmentosum group C (XPC) gene to DNA repair. We stably transfected XPC cells (XP4PA-SV-EB) with XPC cDNA and selected a partially corrected (XP4PA-SE1) and a fully corrected (XP4PA-SE2) clone. Cell survival after UVC (254 nm) exposure was low for XP4PA-SV-EB, intermediate for XP4PA-SE1, and normal for XP4PA-SE2 cells. XP4PA-SV-EB cells had undetectable XPC mRNA and protein levels. XP4PA-SE1 cells had 130% of normal mRNA but 25% of normal protein levels, whereas XP4PA-SE2 cells had an 18-fold mRNA overexpression and normal XPC protein levels compared with normal cells. We measured cyclobutane pyrimidine dimers (CPD) and 6-4 photoproducts (6-4PP) by using specific mAbs and the ELISA technique. XP4PA-SV-EB cells had no detectable removal of CPD or 6-4PP from their global genome by 24 h after 30 J/m(2) UVC exposure. The partially corrected XP4PA-SE1 cells had normal repair of CPD but minimal repair of 6-4PP by 24 h, whereas the fully corrected XP4PA-SE2 cells regained normal CPD and 6-4PP repair capacities. We also exposed pRSVcat plasmid to UVC (to induce CPD and 6-4PP), to UVC + photolyase (to leave only 6-4PP on the plasmid), or to UVB + acetophenone (to induce only CPD). Host cell reactivation of UVB + acetophenone-, but not of UVC + photolyase-treated plasmids was normal in XP4PA-SE1 cells. Thus, increasing XPC gene expression leads to selective repair of CPD in the global genome. Undetectable XPC protein is associated with no repair of CPD or 6-4PP, detectable but subnormal XPC protein levels reconstitute CPD but not 6-4PP repair, and normal XPC protein levels fully reconstitute both CPD and 6-4PP repair. PMID- 10681432 TI - Leptin-deficient (ob/ob) mice are protected from T cell-mediated hepatotoxicity: role of tumor necrosis factor alpha and IL-18. AB - The role of leptin was investigated in two models of T cell-mediated hepatitis: the administration of Con A or of Pseudomonas aeruginosa exotoxin A (PEA). In both models, leptin-deficient (ob/ob) mice were protected from liver damage and showed lower induction of tumor necrosis factor (TNF) alpha and IL-18 compared with their lean littermates. Neutralization of TNF-alpha reduced induction of IL 18 by either Con A (70% reduction) or PEA (40% reduction). Pretreatment of lean mice with either soluble TNF receptors or with an anti-IL-18 antiserum significantly reduced Con A- and PEA-induced liver damage. The simultaneous neutralization of TNF-alpha and IL-18 fully protected the mice against liver toxicity. However, neutralization of either IL-18 or TNF-alpha did not inhibit Con A-induced production of IFN-gamma. Thymus atrophy and alterations in the number of circulating lymphocytes and monocytes were observed in ob/ob mice. Exogenous leptin replacement restored the responsiveness of ob/ob mice to Con A and normalized their lymphocyte and monocyte populations. These results demonstrate that leptin deficiency leads to reduced production of TNF-alpha and IL-18 associated with reduced T cell-mediated hepatotoxicity. In addition, both TNF-alpha and IL-18 appear to be essential mediators of T cell-mediated liver injury. PMID- 10681433 TI - Genetic control of development of the mushroom bodies, the associative learning centers in the Drosophila brain, by the eyeless, twin of eyeless, and Dachshund genes. AB - Mushroom bodies (MBs) are the centers for olfactory associative learning and elementary cognitive functions in the Drosophila brain. By high-resolution neuroanatomy, we show that eyeless (ey), twin of eyeless, and dachshund (dac), which are implicated in eye development, also are expressed in the developing MBs. Mutations of ey completely disrupted the MB neuropils, and a null mutation of dac resulted in marked disruption and aberrant axonal projections. Genetic analyses demonstrated that, whereas ey and dac synergistically control the structural development of the MBs, the two genes are regulated independently in the course of MB development. These data argue for a distinct combinatorial code of regulatory genes for MBs as compared with eye development and suggest conserved roles of Pax6 homologs in the genetic programs of the olfactory learning centers of complex brains. PMID- 10681434 TI - APC mutations are sufficient for the growth of early colorectal adenomas. AB - It is not clear whether APC mutations are sufficient for early colorectal adenomas to grow or whether additional mutations at other loci are required. We previously have screened 210 early colorectal adenomas from familial adenomatous polyposis patients for mutations and allelic loss at APC. Here, we determined whether allelic loss at APC had any effect on the nearby alpha-catenin gene. However, loss on 5q in familial adenomatous polyposis adenomas rarely extended as far as alpha-catenin, and no differences in alpha-catenin protein expression were found in tumors that showed loss encompassing both APC and alpha-catenin. We then screened all 210 tumors for mutations at candidate loci other than APC (K-ras, beta-catenin, and allelic loss at 1p33-p35 and 1p36) and for microsatellite instability (MSI). Each of these loci has been implicated previously in early colorectal tumorigenesis. One tumor harbored a beta-catenin mutation and another MSI, but none showed K-ras mutation or allelic loss at 1p33-p35 or 1p36. These data support the following hypotheses derived from sporadic colorectal tumors: beta-catenin mutations are generally an alternative to mutations at APC, MSI is not usually an early phenomenon in colorectal tumorigenesis, and K-ras mutations are more typical of large- and moderate-sized adenomas. Contrary to some previous reports, chromosome 1p allelic loss is infrequent in very early adenomas. APC mutations are generally sufficient for colorectal tumors to grow to about 1-cm diameter, although chance mutations at other loci can provide these early colorectal adenomas with a selective advantage, and some colorectal tumors may develop along a pathway not involving APC. PMID- 10681435 TI - Cortical correlates of learning in monkeys adapting to a new dynamical environment. AB - In this paper, we describe the neural changes observed in the primary motor cortex of two monkeys while they learned a new motor skill. The monkeys had to adapt their reaching movements to external forces that interfered with the execution of their arm movements. We found a sizable population of cells that changed their tuning properties during exposure to the force field. These cells took on the properties of neurons that are involved in the control of movement. Furthermore, the cells maintained the acquired activity as the monkey readapted to the no-force condition. Recent imaging studies in humans have reported the effects of motor learning in the primary motor cortex. Our results are consistent with the findings of these studies and provide evidence for single-cell plasticity in the primary motor cortex of primates. PMID- 10681436 TI - Toward Anopheles transformation: Minos element activity in anopheline cells and embryos. AB - The ability of the Minos transposable element to function as a transformation vector in anopheline mosquitoes was assessed. Two recently established Anopheles gambiae cell lines were stably transformed by using marked Minos transposons in the presence of a helper plasmid expressing transposase. The markers were either the green fluorescent protein or the hygromycin B phosphotransferase gene driven by the Drosophila Hsp70 promoter. Cloning and sequencing of the integration sites demonstrated that insertions in the cell genome occurred through the action of Minos transposase. Furthermore, an interplasmid transposition assay established that Minos transposase is active in the cytoplasmic environment of Anopheles stephensi embryos: interplasmid transposition events isolated from injected preblastoderm embryos were identified as Minos transposase-mediated integrations, and no events were recorded in the absence of an active transposase. These results demonstrate that Minos vectors are suitable candidates for germ-line transformation of anopheline mosquitoes. PMID- 10681437 TI - Identification of a gene at 11q23 encoding a guanine nucleotide exchange factor: evidence for its fusion with MLL in acute myeloid leukemia. AB - We have identified a gene at 11q23, telomeric to MLL, that encodes a guanine nucleotide exchange factor (GEF). This gene is transcribed into a 9.5-kb mRNA containing a 4.6-kb ORF. By Northern analysis, it was found to be expressed in all human tissues examined including peripheral blood leukocytes, spleen, prostate, testis, ovary, small intestine, colon, and minimally in thymus. Analysis of the predicted protein sequence indicates that it has strong homology to several members of the family of Rho GEFs that includes such oncogenes as Dbl, Vav, Tiam, and Bcr. A patient with primary acute myeloid leukemia (AML) and a karyotype of 51,XY,+8,+19,+3mar was found to have the 5' end of MLL at exon 6 fused in-frame with the 3' end of almost the entire ORF of this gene, which we named LARG for leukemia-associated Rho GEF. Transcriptional orientation of both genes at 11q23 is from centromere to telomere, consistent with other data that suggest the MLL-LARG fusion resulted from an interstitial deletion rather than a balanced translocation. LARG does not appear to have any homology with other MLL partner genes reported thus far. Thus, LARG represents an additional member of the GEF family and a novel MLL fusion partner in acute myeloid leukemia. PMID- 10681438 TI - An artificial transcriptional activating region with unusual properties. AB - We describe a series of transcriptional activators generated by adding amino acids (eight in one case, six in another) to fragments of the yeast Saccharomyces cerevisiae activator Gal4 that dimerize and bind DNA. One of the novel activating regions identified by this procedure is unusual, compared with previously characterized yeast activating regions, in the following ways: it works more strongly than does Gal4's natural activating region as assayed in yeast; it is devoid of acidic residues; and several lines of evidence suggest that it sees targets in the yeast transcriptional machinery at least partially distinct from those seen by Gal4's activating region. PMID- 10681439 TI - IL-18 binding protein increases spontaneous and IL-1-induced prostaglandin production via inhibition of IFN-gamma. AB - IL-18 shares with IL-1 the same family of receptors and several identical signal transduction pathways. Because of these similarities, IL-18 was investigated for its ability to induce prostaglandin E(2) (PGE(2)) synthesis in human peripheral blood mononuclear cells (PBMC), a prominent, proinflammatory property of IL-1. IL 18 was highly active in PBMC by inducing the synthesis of the chemokine IL-8; however, no induction of PGE(2) synthesis nor cyclooxygenase type-2 gene expression was observed in PBMC stimulated with IL-18. In the same cultures, IL 1beta induced a 12-fold increase in PGE(2). Although IL-1beta-induced IL-8 synthesis was augmented 3-fold by IL-18, IL-18 suppressed IL-1beta-induced PGE(2) production by 40%. The suppressive effect of IL-18 on PGE(2) production was mediated by interferon (IFN)-gamma because anti-human IFN-gamma-antibody prevented IL-18-induced reduction in PGE(2). Consistent with these observations, IL-12, a known inducer of IFN-gamma, augmented IL-1beta-induced IFN-gamma but suppressed IL-1beta-induced PGE(2) by 75%. IL-18 binding protein (IL-18BP) is a naturally occurring and specific inhibitor of IL-18. When recombinant IL-18BP was added to PBMC cultures, unexpectedly, spontaneous PGE(2) production increased. PGE(2) production was also increased by the addition of IL-18BP to PBMC stimulated with either IL-1beta or IL-12 and also in whole blood cultures stimulated with Staphylococcus epidermidis. These studies demonstrate that IL 18BP decreases endogenous IL-18 activity by reducing IFN-gamma-mediated responses. PMID- 10681440 TI - Low dielectric response in enzyme active site. AB - The kinetics of charge transfer depend crucially on the dielectric reorganization of the medium. In enzymatic reactions that involve charge transfer, atomic dielectric response of the active site and of its surroundings determines the efficiency of the protein as a catalyst. We report direct spectroscopic measurements of the reorganization energy associated with the dielectric response in the active site of alpha-chymotrypsin. A chromophoric inhibitor of the enzyme is used as a spectroscopic probe. We find that water strongly affects the dielectric reorganization in the active site of the enzyme in solution. The reorganization energy of the protein matrix in the vicinity of the active site is similar to that of low-polarity solvents. Surprisingly, water exhibits an anomalously high dielectric response that cannot be described in terms of the dielectric continuum theory. As a result, sequestering the active site from the aqueous environment inside low-dielectric enzyme body dramatically reduces the dielectric reorganization. This reduction is particularly important for controlling the rate of enzymatic reactions. PMID- 10681441 TI - Electrostatic stress in catalysis: structure and mechanism of the enzyme orotidine monophosphate decarboxylase. AB - Orotidine 5'-monophosphate decarboxylase catalyzes the conversion of orotidine 5' monophosphate to uridine 5'-monophosphate, the last step in biosynthesis of pyrimidine nucleotides. As part of a Structural Genomics Initiative, the crystal structures of the ligand-free and the6-azauridine 5'-monophosphate-complexed forms have been determined at 1.8 and 1.5 A, respectively. The protein assumes a TIM-barrel fold with one side of the barrel closed off and the other side binding the inhibitor. A unique array of alternating charges (Lys-Asp-Lys-Asp) in the active site prompted us to apply quantum mechanical and molecular dynamics calculations to analyze the relative contributions of ground state destabilization and transition state stabilization to catalysis. The remarkable catalytic power of orotidine 5'-monophosphate decarboxylase is almost exclusively achieved via destabilization of the reactive part of the substrate, which is compensated for by strong binding of the phosphate and ribose groups. The computational results are consistent with a catalytic mechanism that is characterized by Jencks's Circe effect. PMID- 10681442 TI - The crystal structure and mechanism of orotidine 5'-monophosphate decarboxylase. AB - The crystal structure of Bacillus subtilis orotidine 5'-monophosphate (OMP) decarboxylase with bound uridine 5'-monophosphate has been determined by multiple wavelength anomalous diffraction phasing techniques and refined to an R-factor of 19.3% at 2.4 A resolution. OMP decarboxylase is a dimer of two identical subunits. Each monomer consists of a triosephosphate isomerase barrel and contains an active site that is located across one end of the barrel and near the dimer interface. For each active site, most of the residues are contributed by one monomer with a few residues contributed from the adjacent monomer. The most highly conserved residues are located in the active site and suggest a novel catalytic mechanism for decarboxylation that is different from any previously proposed OMP decarboxylase mechanism. The uridine 5'-monophosphate molecule is bound to the active site such that the phosphate group is most exposed and the C5 C6 edge of the pyrimidine base is most buried. In the proposed catalytic mechanism, the ground state of the substrate is destabilized by electrostatic repulsion between the carboxylate of the substrate and the carboxylate of Asp60. This repulsion is reduced in the transition state by shifting negative charge from the carboxylate to C6 of the pyrimidine, which is close to the protonated amine of Lys62. We propose that the decarboxylation of OMP proceeds by an electrophilic substitution mechanism in which decarboxylation and carbon-carbon bond protonation by Lys62 occur in a concerted reaction. PMID- 10681443 TI - Transgenic mice neuronally expressing baculoviral p35 are resistant to diverse types of induced apoptosis, including seizure-associated neurodegeneration. AB - Apoptosis is a cell-suicide process that appears to play a central role not only during normal neuronal development but also in several neuropathological disease states. An important component of this process is a proteolytic cascade involving a family of cysteine proteases called caspases. Caspase inhibitors have been demonstrated to be effective in inhibiting neuronal cell death in various apoptotic paradigms. We have created transgenic mice that neuronally express the baculoviral caspase inhibitor p35. Neuronal expression of the p35 protein was found to confer functional caspase inhibitory activity and prevent apoptosis in isolated cerebellar granular cultures induced to undergo apoptosis either via staurosporine treatment or through withdrawal of extracellular potassium. Neuronal expression of p35 was also found to attenuate neurodegeneration associated with the excitotoxic glutamate analogue kainic acid (KA) in vitro and in vivo. Organotypic hippocampal cultures isolated from p35 transgenics demonstrated lowered caspase activity and decreased apoptosis compared with wild type when exposed to KA. In vivo injection of KA also produced decreased caspase activity and cell death in p35 transgenics vs. wild type. These results suggest that the presence of p35 in neurons in vivo is protective against various types of apoptosis, including seizure-related neurodegeneration, and that caspases may be attractive potential targets for preventing neuronal injury associated with diseases such as epilepsy. These mice also provide a valuable tool for exploring the role of caspases in other neuropathological conditions in which apoptosis has been implicated. PMID- 10681444 TI - A critical residue for isoform difference in tetrodotoxin affinity is a molecular determinant of the external access path for local anesthetics in the cardiac sodium channel. AB - Membrane-impermeant quaternary derivatives of lidocaine (QX222 and QX314) block cardiac Na(+) channels when applied from either side of the membrane, but they block neuronal and skeletal muscle channels poorly from the outside. To find the molecular determinants of the cardiac external QX access path, mutations of adult rat skeletal muscle (micro1) and rat heart (rH1) Na(+) channels were studied by two-electrode voltage clamp in Xenopus oocytes. Mutating the micro1 domain I P loop Y401, which is the critical residue for isoform differences in tetrodotoxin block, to the heart sequence (Y401C) allowed outside QX222 block, but its mutation to brain type (Y401F) showed little block. mu1-Y401C accelerated recovery from block by internal QX222. Block by external QX222 in mu1-Y401C was diminished by chemical modification with methanethiosulfonate ethylammonium (MTSEA) to the outer vestibule or by a double mutant (mu1-Y401C/F1579A), which altered the putative local anesthetic binding site. The reverse mutation in heart rH1-C374Y reduced outside QX314 block and slowed dissociation of internal QX222. Mutation of mu1-C1572 in IVS6 to Thr, the cardiac isoform residue (C1572T), allowed external QX222 block, and accelerated recovery from internal QX222 block, as reported. Blocking efficacy of outside QX222 in mu1-Y401C was more than that in mu1-C1572T, and the double mutant (mu1-Y401C/C1572T) accelerated internal QX recovery more than mu1-Y401C or mu1-C1572T alone. We conclude that the isoform specific residue (Tyr/Phe/Cys) in the P-loop of domain I plays an important role in drug access as well as in tetrodotoxin binding. Isoform-specific residues in the IP-loop and IVS6 determine outside drug access to an internal binding site. PMID- 10681445 TI - Compartmentation protects trypanosomes from the dangerous design of glycolysis. AB - Unlike in other organisms, in trypanosomes and other Kinetoplastida the larger part of glycolysis takes place in a specialized organelle, called the glycosome. At present it is impossible to remove the glycosome without changing much of the rest of the cell. It would seem impossible, therefore, to assess the metabolic consequences of this compartmentation. Therefore, we here develop a computer experimentation approach, which we call computational cell biology. A validated molecular kinetic computer replica was built of glycolysis in the parasite Trypanosoma brucei. Removing the glycosome membrane in that replica had little effect on the steady-state flux, which argues against the prevalent speculation that glycosomes serve to increase flux by concentrating the enzymes. Removal of the membrane did cause (i) the sugar phosphates to rise to unphysiologically high levels, which must have pathological effects, and (ii) a failure to recover from glucose deprivation. We explain these effects on the basis of the biochemical organization of the glycosome. We conclude (i) that the glycosome protects trypanosomes from the negative side effects of the "turbo" structure of glycolysis and (ii) that computer experimentation based on solid molecular data is a powerful tool to address questions that are not, or not yet, accessible to experimentation. PMID- 10681446 TI - A spectroscopic method for observing the domain movement of the Rieske iron sulfur protein. AB - The g-tensor orientation of the chemically reduced Rieske cluster in cytochrome bc(1) complex from Rhodovulum sulfidophilum with respect to the membrane was determined in the presence and absence of inhibitors and in the presence of oxidized and reduced quinone in the quinol-oxidizing-site (Q(o)-site) by EPR on two-dimensionally ordered samples. Almost identical orientations were observed when oxidized or reduced quinone, stigmatellin, or 5-(n-undecyl)-6-hydroxy-4,7 dioxobenzothiazole was present. Occupancy of the Q(o)-site by myxothiazole induced appearance of a minority population with a substantially differing conformation and presence of E-beta-methoxyacrylate-stilbene significantly reduced the contribution of the major conformation observed in the other cases. Furthermore, when the oxidized iron-sulfur cluster was reduced at cryogenic temperatures by the products of radiolysis, the orientation of its magnetic axes was found to differ significantly from that of the chemically reduced center. The "irradiation-induced" conformation converts to that of the chemically reduced center after thawing of the sample. These results confirm the effects of Q(o) site inhibitors on the equilibrium conformation of the Rieske iron-sulfur protein and provide evidence for a reversible redox-influenced interconversion between conformational states. Moreover, the data obtained with the iron-sulfur protein demonstrate that the conformation of "EPR-inaccessible" reduction states of redox centers can be studied by inducing changes of redox state at cryogenic temperatures. This technique appears applicable to a wide range of comparable electron transfer systems performing redox-induced conformational changes. PMID- 10681447 TI - The accuracy of codon recognition by polypeptide release factors. AB - The precision with which individual termination codons in mRNA are recognized by protein release factors (RFs) has been measured and compared with the decoding of sense codons by tRNA. An Escherichia coli system for protein synthesis in vitro with purified components was used to study the accuracy of termination by RF1 and RF2 in the presence or absence of RF3. The efficiency of factor-dependent termination at all sense codons differing from any of the three stop codons by a single mutation was measured and compared with the efficiency of termination at the three stop codons. RF1 and RF2 discriminate against sense codons related to stop codons by between 3 and more than 6 orders of magnitude. This high level of accuracy is obtained without energy-driven error correction (proofreading), in contrast to codon-dependent aminoacyl-tRNA recognition by ribosomes. Two codons, UAU and UGG, stand out as hotspots for RF-dependent premature termination. PMID- 10681448 TI - Association of SWAP-70 with the B cell antigen receptor complex. AB - SWAP-70 is a component of an enzyme complex that recombines Ig switch regions in vitro. We report here the cloning of the human cDNA and its B lymphocyte-specific expression. Although its sequence contains three nuclear localization signals, in small resting B cells, SWAP-70 is mainly found in the cytoplasm. On stimulation, SWAP-70 translocates to the nucleus. In activated, class-switching B cell cultures, it is associated with membrane IgG, but not IgM. The membrane Ig association requires a functional pleckstrin homology domain and is controlled by the C terminus. We suggest that SWAP-70 is involved not only in nuclear events but also in signaling in B cell activation. PMID- 10681449 TI - Noise-based switches and amplifiers for gene expression. AB - The regulation of cellular function is often controlled at the level of gene transcription. Such genetic regulation usually consists of interacting networks, whereby gene products from a single network can act to control their own expression or the production of protein in another network. Engineered control of cellular function through the design and manipulation of such networks lies within the constraints of current technology. Here we develop a model describing the regulation of gene expression and elucidate the effects of noise on the formulation. We consider a single network derived from bacteriophage lambda and construct a two-parameter deterministic model describing the temporal evolution of the concentration of lambda repressor protein. Bistability in the steady-state protein concentration arises naturally, and we show how the bistable regime is enhanced with the addition of the first operator site in the promotor region. We then show how additive and multiplicative external noise can be used to regulate expression. In the additive case, we demonstrate the utility of such control through the construction of a protein switch, whereby protein production is turned "on" and "off" by using short noise pulses. In the multiplicative case, we show that small deviations in the transcription rate can lead to large fluctuations in the production of protein, and we describe how these fluctuations can be used to amplify protein production significantly. These results suggest that an external noise source could be used as a switch and/or amplifier for gene expression. Such a development could have important implications for gene therapy. PMID- 10681450 TI - Vitamin E reduces chromosomal damage and inhibits hepatic tumor formation in a transgenic mouse model. AB - We have previously shown that chronic activation of mitogenic signaling induced by over-expression of c-myc and transforming growth factor-alpha (TGFalpha) transgenes in mouse liver induces a state of oxidative stress. We therefore proposed that increased reactive oxygen species (ROS) generation might be responsible for the extensive chromosomal damage and acceleration of hepatocarcinogenesis characteristic for TGFalpha/c-myc mice. In this study, we show that vitamin E (VE), a potent free radical scavenging antioxidant, is able to protect liver tissue against oxidative stress and suppress tumorigenic potential of c-myc oncogene. Dietary supplementation with VE, starting from weaning, decreased ROS generation coincident with a marked inhibition of hepatocyte proliferation while increasing the chromosomal as well as mtDNA stability in the liver. Similarly, dietary VE reduced liver dysplasia and increased viability of hepatocytes. At 6 mo of age, VE treatment decreased the incidence of adenomas by 65% and prevented malignant conversion. These results indicate that ROS generated by over-expression of c-myc and TGFalpha in the liver are the primary carcinogenic agents in this animal model. Furthermore, the data demonstrate that dietary supplementation of VE can effectively inhibit liver cancer development. PMID- 10681451 TI - Meiotic instability of CAG repeat tracts occurs by double-strand break repair in yeast. AB - Expansion of trinucleotide repeats is associated with a growing number of human diseases. The mechanism and timing of expansion of the repeat tract are poorly understood. In humans, trinucleotide repeats show extreme meiotic instability, and expansion of the repeat tract has been suggested to occur in the germ-line mitotic divisions or postmeiotically during early divisions of the embryo. Studies in model organisms have indicated that polymerase slippage plays a major role in the repeat tract instability and meiotic instability is severalfold higher than the mitotic instability. We show here that meiotic instability of the CAG/CTG repeat tract in yeast is associated with double-strand break (DSB) formation within the repeated sequences, and that the DSB formation is dependent on the meiotic recombination machinery. The DSB repair results in both expansions and contractions of the CAG repeat tract. PMID- 10681452 TI - The hippocampal neurons of neuronal apoptosis inhibitory protein 1 (NAIP1) deleted mice display increased vulnerability to kainic acid-induced injury. AB - The neuronal apoptosis inhibitory protein (NAIP) is a member of a novel family of inhibitor of apoptosis (IAP) proteins. The IAP genes are highly conserved from baculovirus to metazoans and suppress apoptosis induced by a variety of triggers both in vitro and in vivo. Here we describe the generation and characterization of mice with the targeted deletion of NAIP1. We demonstrate that the NAIP1 deleted mice develop normally. However, the survival of pyramidal neurons in the hippocampus after kainic acid-induced limbic seizures is greatly reduced in the NAIP1 knock-out animals. Thus, although NAIP1 is not necessary for normal development of murine central nervous system, the endogenous NAIP1 is required for neuronal survival in pathological conditions. PMID- 10681453 TI - Dendritic cells acquire the MAGE-3 human tumor antigen from apoptotic cells and induce a class I-restricted T cell response. AB - In an attempt to transduce monocyte-derived dendritic cells (DCs) with a retroviral vector coding for an intracytoplasmic tumor antigen (TAA), we were confronted by the evident dissociation between the ability of the treated DCs to induce a TAA-specific response, and the presence of integrated vector proviral DNA. The TAA, i.e., MAGE-3, was acquired by DCs and presented to immune effectors, thanks to the property of DCs to uptake the apoptotic bodies released by the irradiated vector-producing cells. Indeed, we observed that upon irradiation vector-producing cells underwent apoptotic cell death, monitored by annexin V and propidium iodide staining, and were phagocytosed by DCs. Lymphocytes obtained from a patient affected by a MAGE-3(+) melanoma, were stimulated in vitro with autologous DCs previously exposed to irradiated MAGE-3 expressing cells. This procedure led to the induction of MAGE-3-specific cytotoxic effectors, directed against a yet unknown MAGE-3 epitope presented by HLA-A*B5201 molecules. These data demonstrate that DCs can present engulfed human TAAs, thus providing strategies for cancer vaccination. PMID- 10681454 TI - Transferrin receptor 2: continued expression in mouse liver in the face of iron overload and in hereditary hemochromatosis. AB - Hereditary hemochromatosis (HH) is a common autosomal recessive disorder characterized by excess absorption of dietary iron and progressive iron deposition in several tissues, particularly liver. Liver disease resulting from iron toxicity is the major cause of death in HH. Hepatic iron loading in HH is progressive despite down-regulation of the classical transferrin receptor (TfR). Recently a human cDNA highly homologous to TfR was identified and reported to encode a protein (TfR2) that binds holotransferrin and mediates uptake of transferrin-bound iron. We independently identified a full-length murine EST encoding the mouse orthologue of the human TfR2. Although homologous to murine TfR in the coding region, the TfR2 transcript does not contain the iron responsive elements found in the 3' untranslated sequence of TfR mRNA. To determine the potential role for TfR2 in iron uptake by liver, we investigated TfR and TfR2 expression in normal mice and murine models of dietary iron overload (2% carbonyl iron), dietary iron deficiency (gastric parietal cell ablation), and HH (HFE -/-). Northern blot analyses demonstrated distinct tissue-specific patterns of expression for TfR and TfR2, with TfR2 expressed highly only in liver where TfR expression is low. In situ hybridization demonstrated abundant TfR2 expression in hepatocytes. In contrast to TfR, TfR2 expression in liver was not increased in iron deficiency. Furthermore, hepatic expression of TfR2 was not down-regulated with dietary iron loading or in the HFE -/- model of HH. From these observations, we propose that TfR2 allows continued uptake of Tf-bound iron by hepatocytes even after TfR has been down-regulated by iron overload, and this uptake contributes to the susceptibility of liver to iron loading in HH. PMID- 10681455 TI - The protease thrombin is an endogenous mediator of hippocampal neuroprotection against ischemia at low concentrations but causes degeneration at high concentrations. AB - We have considered the extracellular serine protease thrombin and its receptor as endogenous mediators of neuronal protection against brain ischemia. Exposure of gerbils to prior mild ischemic insults, here two relatively short-lasting occlusions (2 min) of both common carotid arteries applied at 1-day intervals 2 days before a severe occlusion (6 min), caused a robust ischemic tolerance of hippocampal CA1 neurons. This resistance was impaired if the specific thrombin inhibitor hirudin was injected intracerebroventricularly before each short lasting insult. Thus, efficient native neuroprotective mechanisms exist and endogenous thrombin seems to be involved therein. In vitro experiments using organotypic slice cultures of rat hippocampus revealed that thrombin can have protective but also deleterious effects on hippocampal CA1 neurons. Low concentrations of thrombin (50 pM, 0.01 unit/ml) or of a synthetic thrombin receptor agonist (10 microM) induced significant neuroprotection against experimental ischemia. In contrast, 50 nM (10 units/ml) thrombin decreased further the reduced neuronal survival that follows the deprivation of oxygen and glucose, and 500 nM even caused neuronal cell death by itself. Degenerative thrombin actions also might be relevant in vivo, because hirudin increased the number of surviving neurons when applied before a 6-min occlusion. Among the thrombin concentrations tested, 50 pM induced intracellular Ca(2+) spikes in fura 2-loaded CA1 neurons whereas higher concentrations caused a sustained Ca(2+) elevation. Thus, distinct Ca(2+) signals may define whether or not thrombin initiates protection. Taken together, in vivo and in vitro data suggest that thrombin can determine neuronal cell death or survival after brain ischemia. PMID- 10681456 TI - Representation of actions in rats: the role of cerebellum in learning spatial performances by observation. AB - Experimental evidence demonstrates that cerebellar networks are involved in spatial learning, controlling the acquisition of exploration strategies without blocking motor execution of the task. Action learning by observation has been considered somehow related to motor physiology, because it provides a way of learning performances that is almost as effective as the actual execution of actions. Neuroimaging studies demonstrate that observation of movements performed by others, imagination of actions, and actual execution of motor performances share common neural substrates and that the cerebellum is among these shared areas. The present paper analyzes the effects of observation in learning a spatial task, focusing on the cerebellar role in learning a spatial ability through observation. We allowed normal rats to observe 200 Morris water maze trials performed by companion rats. After this observation training, "observer" rats underwent a hemicerebellectomy and then were tested in the Morris water maze. In spite of the cerebellar lesion, they displayed no spatial defects, exhibiting exploration abilities comparable to controls. When the cerebellar lesion preceded observation training, a complete lack of spatial observational learning was observed. Thus, as demonstrated already for the acquisition of spatial procedures through actual execution, cerebellar circuits appear to play a key role in the acquisition of spatial procedures also through observation. In conclusion, the present results provide strong support for a common neural basis in the observation of actions that are to be reproduced as well as in the actual production of the same actions. PMID- 10681457 TI - Oral administration of a dual analog of two myasthenogenic T cell epitopes down regulates experimental autoimmune myasthenia gravis in mice. AB - Myasthenia gravis (MG) and experimental autoimmune MG (EAMG) are T cell regulated, antibody-mediated autoimmune diseases. The major autoantigen in MG is the nicotinic acetylcholine receptor (AChR). Two peptides, representing sequences of the human AChR alpha-subunit, p195-212 and p259-271, were previously shown to be immunodominant T cell epitopes in MG patients as well as, respectively, in SJL and BALB/c mice. A dual analog (termed Lys-262-Ala-207) composed of the tandemly arranged two single amino acid analogs of p195-212 and p259-271 was shown to inhibit, in vitro and in vivo, MG-associated autoimmune responses. Furthermore, the dual analog could down-regulate myasthenogenic manifestations in mice with EAMG that was induced by inoculation of a pathogenic T cell line. In the present study, the ability of the dual analog to treat EAMG induced in susceptible C57BL/6 mice by native Torpedo AChR was evaluated. Mice that were diagnosed to have clinical symptoms of EAMG were treated with the dual analog by oral administration, 500 microg per mouse three times a week for 5-8 weeks. Treatment with the dual analog down-regulated the clinical manifestations of the ongoing disease as assessed by the clinical score, grip strength (measured by a grip strength meter), and electromyography. The effects on the clinical EAMG correlated with a reduced production of anti-AChR antibody as well as a decrease in the secretion of interleukin-2 and, more dramatically, interferon-gamma, in response to AChR triggering. Thus, the dual analog is an efficient immunomodulator of EAMG in mice and might be of specific therapeutic potential for MG. PMID- 10681458 TI - Molecular mobility in the cytoplasm: an approach to describe and predict lifespan of dry germplasm. AB - Molecular mobility is increasingly considered a key factor influencing storage stability of biomolecular substances, because it is thought to control the rate of detrimental reactions responsible for reducing the shelf life of, for instance, pharmaceuticals, food, and germplasm. We investigated the relationship between aging rates of germplasm and the rotational motion of a polar spin probe in the cytoplasm under different storage conditions using saturation transfer electron paramagnetic resonance spectroscopy. Rotational motion of the spin probe in the cytoplasm of seed and pollen of various plant species changed as a function of moisture content and temperature in a manner similar to aging rates or longevity. A linear relationship was established between the logarithms of rotational motion and aging rates or longevity. This linearity suggests that detrimental aging rates are associated with molecular mobility in the cytoplasm. By measuring the rotational correlation times at low temperatures at which experimental determination of longevity is practically impossible, this linearity enabled us to predict vigor loss or longevity. At subzero temperatures, moisture contents for maximum life span were predicted to be higher than those hitherto used in genebanks, urging for a reexamination of seed storage protocols. PMID- 10681459 TI - Engineered herpes simplex virus expressing IL-12 in the treatment of experimental murine brain tumors. AB - Genetically engineered, neuroattenuated herpes simplex viruses (HSVs) expressing various cytokines can improve survival when used in the treatment of experimental brain tumors. These attenuated viruses have both copies of gamma(1)34.5 deleted. Recently, we demonstrated increased survival of C57BL/6 mice bearing syngeneic GL 261 gliomas when treated with an engineered HSV expressing IL-4, as compared with treatment with the parent construct (gamma(1)34. 5(-)) alone or with a virus expressing IL-10. Herein, we report construction of a conditionally replication competent mutant expressing both subunits of mIL-12 (M002) and its evaluation in a syngeneic neuroblastoma murine model. IL-12 induces a helper T cell subset type 1 response, which may induce more durable antitumor effects. In vitro studies showed that, when infected with M002, both Vero cells and murine Neuro-2a neuroblastoma cells produced physiologically relevant levels of IL-12 heterodimers, as determined by ELISA. M002 was cytotoxic for Neuro-2a cells and human glioma cell lines U251MG and D54MG. Neurotoxicity studies, as defined by plaque-forming units/LD(50), performed in HSV-1-sensitive A/J strain mice found that M002 was not toxic even at high doses. When evaluated in an intracranial syngeneic neuroblastoma murine model, median survival of M002-treated animals was significantly longer than the median survival of animals treated with R3659, the parent gamma(1)34.5(-) mutant lacking any cytokine gene insert. Immunohistochemical analysis of M002-treated tumors identified a pronounced influx of CD4(+) T cells and macrophages as well as CD8(+) cells when compared with an analysis of R3659-treated tumors. We conclude that M002 produced a survival benefit via oncolytic effects combined with immunologic effects meditated by helper T cells of subset type 1. PMID- 10681460 TI - Patterned deposition of cells and proteins onto surfaces by using three dimensional microfluidic systems. AB - Three-dimensional microfluidic systems were fabricated and used to pattern proteins and mammalian cells on a planar substrate. The three-dimensional topology of the microfluidic network in the stamp makes this technique a versatile one with which to pattern multiple types of proteins and cells in complex, discontinuous structures on a surface. The channel structure, formed by the stamp when it is in contact with the surface of the substrate, limits migration and growth of cells in the channels. With the channel structure in contact with the surface, the cells stop dividing once they form a confluent layer. Removal of the stamp permits the cells to spread and divide. PMID- 10681461 TI - Loss of brain-derived neurotrophic factor-dependent neural crest-derived sensory neurons in neurotrophin-4 mutant mice. AB - Peripheral ganglion neurons confer sensory information including touch, pain, temperature, and proprioception. Sensory modality is linked to specific neurotrophin (NTF) requirements. NT-3 supports survival of neurons that differentiate primarily into proprioceptors whereas nerve growth factor and brain derived neurotrophic factor (BDNF) support subpopulations that transmit nociception and mechanoreception, respectively. We examined sensory neurons of gene-targeted mouse mutants at the NT-4, BDNF, NT-3, and TrkA loci. We show that NT-4 functions early in gangliogenesis, upstream of BDNF. In the absence of NT-4 function, BDNF-dependent, TrkB-expressing neurons fail to appear. The results are consistent with the model that precursor cells intended to become BDNF-dependent mechanoreceptors instead differentiate into NT-3-dependent proprioceptive neurons. PMID- 10681462 TI - Physical contact between lipopolysaccharide and toll-like receptor 4 revealed by genetic complementation. AB - Some mammalian species show an ability to discriminate between different lipopolysaccharide (LPS) partial structures (for example, lipid A and its congener LA-14-PP, which lacks secondary acyl chains), whereas others do not. Using a novel genetic complementation system involving the transduction of immortalized macrophages from genetically unresponsive C3H/HeJ mice, we now have shown that the species-dependent discrimination between intact LPS and tetra-acyl LPS partial structures is fully attributable to the species origin of Toll-like receptor 4 (Tlr4), an essential membrane-spanning component of the mammalian LPS sensor. Because Tlr4 interprets the chemical structure of an LPS molecule, we conclude that LPS must achieve close physical proximity with Tlr4 in the course of signal transduction. PMID- 10681463 TI - The cellulose synthase gene of Dictyostelium. AB - Cellulose is a major component of the extracellular matrices formed during development of the social amoeba, Dictyostelium discoideum. We isolated insertional mutants that failed to accumulate cellulose and had no cellulose synthase activity at any stage of development. Development proceeded normally in the null mutants up to the beginning of stalk formation, at which point the culminating structures collapsed onto themselves, then proceeded to attempt culmination again. No spores or stalk cells were ever made in the mutants, with all cells eventually lysing. The predicted product of the disrupted gene (dcsA) showed significant similarity to the catalytic subunit of cellulose synthases found in bacteria. Enzyme activity and normal development were recovered in strains transformed with a construct expressing the intact dcsA gene. Growing amoebae carrying the construct accumulated the protein product of dcsA, but did not make cellulose until they had developed for at least 10 hr. These studies show directly that the product of dcsA is necessary, but not sufficient, for synthesis of cellulose. PMID- 10681464 TI - Arabidopsis cytochrome P450s that catalyze the first step of tryptophan-dependent indole-3-acetic acid biosynthesis. AB - Plants synthesize numerous secondary metabolites that are used as developmental signals or as defense against pathogens. Tryptophan (Trp)-derived secondary metabolites include camalexin, indole glucosinolates, and indole-3-acetic acid (IAA); however, the steps in their synthesis from Trp or its precursors remain unclear. We have identified two Arabidopsis cytochrome P450s (CYP79B2 and CYP79B3) that can convert Trp to indole-3-acetaldoxime (IAOx), a precursor to IAA and indole glucosinolates. PMID- 10681466 TI - Protein folding and unfolding on a complex energy landscape. AB - Recent theories of protein folding suggest that individual proteins within a large ensemble may follow different routes in conformation space from the unfolded state toward the native state and vice versa. Herein, we introduce a new type of kinetics experiment that shows how different unfolding pathways can be selected by varying the initial reaction conditions. The relaxation kinetics of the major cold shock protein of Escherichia coli (CspA) in response to a laser induced temperature jump are exponential for small temperature jumps, indicative of folding through a two-state mechanism. However, for larger jumps, the kinetics become strongly nonexponential, implying the existence of multiple unfolding pathways. We provide evidence that both unfolding across an energy barrier and diffusive downhill unfolding can occur simultaneously in the same ensemble and provide the experimental requirements for these to be observed. PMID- 10681467 TI - Defensive use of a fecal thatch by a beetle larva (Hemisphaerota cyanea). AB - The larva of the tortoise beetle, Hemisphaerota cyanea (Chrysomelidae, Cassidinae), constructs a thatch from long filamentous fecal strands, beneath which it is totally concealed. The thatch is not discarded at molting but is enlarged by addition of strands as the larva grows. Thatch construction begins when the larva hatches from the egg. Pupation occurs beneath the thatch. Two predators, a coccinellid beetle larva (Cycloneda sanguinea) and a pentatomid bug (Stiretrus anchorago), were shown to be thwarted by the thatch. However, one predator, a carabid beetle (Calleida viridipennis), feeds on the larva by either forcing itself beneath the thatch or chewing its way into it. The attack behavior is stereotyped, suggesting that the beetle feeds on Hemisphaerota larvae as a matter of routine. PMID- 10681465 TI - The gene coding for the Hrp pilus structural protein is required for type III secretion of Hrp and Avr proteins in Pseudomonas syringae pv. tomato. AB - Bacterial surface appendages called pili often are associated with DNA and/or protein transfer between cells. The exact function of pili in the transfer process is not understood and is a matter of considerable debate. The Hrp pilus is assembled by the Hrp type III protein secretion system of Pseudomonas syringae pv. tomato (Pst) strain DC3000. In this study, we show that the hrpA gene, which encodes the major subunit of the Hrp pilus, is required for secretion of putative virulence proteins, such as HrpW and AvrPto. In addition, the hrpA gene is required for full expression of genes that encode regulatory, secretion, and effector proteins of the type III secretion system. hrpA-mediated gene regulation apparently is through effect on the mRNA level of two previously characterized regulatory genes, hrpR and hrpS. Ectopic expression of the hrpRS gene operon restored gene expression, but not protein secretion, in the hrpA mutant. Three single amino acid mutations at the HrpA carboxyl terminus were identified that affect the secretion or regulatory function of the HrpA protein. These results define an essential role of the Hrp pilus structural gene in protein secretion and coordinate regulation of the type III secretion system in Pst DC3000. PMID- 10681469 TI - Debeerius ellefseni (Fam. Nov., Gen. Nov., Spec. Nov.), an autodiastylic chondrichthyan from the Mississippian bear gulch limestone of Montana (USA), the relationships of the chondrichthyes, and comments on gnathostome evolution. AB - Debeerius ellefseni is an autodiastylic, operculate chondrichthyan from the 320 million-year-old Bear Gulch limestone (Heath Formation, Big Snowy Group, Upper Chesterian) of Montana, USA. Cranial and postcranial morphologies show strong affinities to the holocephalan cochliodonts and Chimaeriformes. The heterodont dentition is, however, selachian in plan. Debeerius ellefseni's cranial, postcranial, and suspensorial characters identify this fish as a paraselachian, an early chondrichthyan with a morphology intermediate to the chimaeroid and selachian plans. They also support the division of Chondrichthyes into the subclasses Elasmobranchii and Euchondrocephali (Paraselachii + Holocephalimorpha). Details of the anatomy of D. ellefseni are reviewed in light of recent advances in understanding vertebrate splanchnocranial development and, thus, permit a discussion of historically problematic craniate features, including labial cartilages and the nature of the mandibular arch relative to hyoid and branchial arches. Developmental and evolutionary considerations of these characters are consistent with an embryonic body plan shared by both lampreys and gnathostomes. Debeerius ellefseni's suspensorium corresponds to the plesiomorphous gnathostome condition theorized by DeBeer and Moy-Thomas in 1935. The description of this autodiastylic condition is clarified to include observations of the hyoid arch, which is complete with a pharyngohyal and provides support for the primary opercular valve. The confirmation of an autodiastylic suspensorium requires a reexamination of the commonly accepted paradigm for jaw evolution. The selachian, chimaeroid, and actinopterygian conditions are all derivable from this plesiomorphous state; the placoderm and sarcopterygian conditions are related and probably similarly derived. The comparable osteichthyan suspensorium is best represented by the suspensorial condition of coelacanths. PMID- 10681468 TI - A protein-based therapeutic for human cytomegalovirus infection. AB - Current antiviral strategies target viral gene products. Although initially successful, their severe toxicity and susceptibility to circumvention by the generation of drug-resistant variants limit their usefulness. By contrast, the central role of the host cell serine endoprotease furin in the proteolytic activation of numerous pathogens points to the endoprotease as a strategic target for therapeutics. Herein, we show that the production of infectious human cytomegalovirus is dramatically reduced by exogenous addition of a bioengineered serpin, alpha(1)-PDX. This protein is a potent and selective furin inhibitor (K(i) = 0.6 nM) and is 10-fold more effective than currently used antiherpetic agents in cell-culture models. The requirement of furin for the processing of envelope glycoproteins from many pathogenic viruses and for the activation of several bacterial toxins suggests that selective inhibitors of furin have potential as broad-based anti-pathogens. PMID- 10681470 TI - Descriptive study of the diaphragm and lungs in the short-nosed echidna, Tachyglossus aculeatus (Mammalia: monotremata). AB - In this descriptive study, we characterize the diaphragm and lungs of the short nosed echidna, Tachyglossus aculeatus, using a combination of gross anatomical, light-microscopic, electron microscopic, and morphometric techniques, including airway casting. The diaphragm is inclined from ventro-cranial to dorso-caudal and possesses a large central tendon (centrum tendineum). The crural and costal muscle groups and the associated trigoni are located in the same positions as in other mammals. The bronchial branching pattern reveals cranially broad, tapering stem bronchi and an unusually small number of first order bronchi. The asymmetrical primary branching pattern and possibly also the asymmetry of right and left lungs are plesiomorphic within the Mammalia. The histology and ultrastructure of the airways and lung parenchyma reveal no unusual features: alveolar type 1 and type 2 cells in the parenchyma; type 2 cells, exocrine bronchiolar cells (Clara cells), ciliated cells, and goblet cells in the terminal airways and the latter two cell types in the bronchi. Both a double and a single capillary net are found on the interalveolar septa. The high capillary loading of the double net may be of selective advantage because of long apneas and low metabolic rate in the echidna. PMID- 10681471 TI - Glycoconjugate sugar residues in the chick embryo developing lung: a lectin histochemical study. AB - A lectin histochemical study was performed to investigate the distribution and changes of the oligosaccharidic component of the glycoconjugates in the lung of chick embryos, of 1-day-old chick, and of the adult animal. For this purpose, a battery of seven horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, Con A, LTA, and UEA I) were employed. During the first phase of parabronchi and atria formation, D-galactose-(beta1-->3)-N-acetyl-D-galactosamine, beta-N-acetyl D-galactosamine, D-glucosamine, alpha-D-mannose, and sialic acid, present at the level of the surface and of cytoplasmic granules of the lining epithelial cells, seem to play a role in regulating morphogenetic phenomena. In the subsequent phases, the parabronchial lumen and the atrial cavities were characterized by the presence of lectin-reactive material rich in terminal D-galactose-(beta1-->3)-N acetyl-D-galactosamine, beta-N-acetyl-D-galactosamine, D-glucosamine and alpha-D mannose. From day 18 onwards and immediately after hatching, the free border of the cells lining the air capillaries was characterized by the presence of beta-N acetyl-D-galactosamine and alpha-D-mannose. The appearance of these sugar residues was concomitant with the beginning of respiratory activity. PMID- 10681472 TI - Ovarian germinal epithelium and folliculogenesis in the common snook, Centropomus undecimalis (Teleostei: centropomidae). AB - The ovarian germinal epithelium in the common snook, Centropomus undecimalis, is described. It consists of epithelial and prefollicle cells that surround germ cells, either oogonia or oocytes, respectively. The germinal epithelium borders a body cavity, the ovarian lumen, and is supported by a basement membrane that also separates the epithelial compartment of the ovarian lamellae from the stromal compartment. During folliculogenesis, the epithelial cells, whose cytoplasmic processes encompass meiotic oocytes, transform into prefollicle cells, which become follicle cells at the completion of folliculogenesis. The follicle is a derivative of the germinal epithelium and is composed of the oocyte and surrounding follicle cells. It is separated from the encompassing theca by a basement membrane. The cells that form the theca interna are derived from prethecal cells within the extravascular space of the ovarian stroma. The theca externa differentiates from undifferentiated cells within the stromal compartment of the ovary, from within the extravascular space. The theca interna and the theca externa are not considered to be part of the follicle and are derived from a different ovarian compartment than the follicle. Meiosis commences while oocytes are still within the germinal epithelium and proceeds as far as arrested diplotene of the first meiotic prophase. The primary growth phase of oocyte development also begins while oocytes are still within the germinal epithelium or attached to it in a cell nest. The definitions used herein are consistent between sexes and with the mammalian literature. PMID- 10681473 TI - Renal anatomy in sparrows from different environments. AB - The renal anatomy of three species of sparrows, two from mesic areas, the House Sparrow (Passer domesticus) and Song Sparrow (Melospiza melodia), and one salt marsh species, the Savannah Sparrow (Passerculus sandwichensis) was examined. Electron microscopy was used to describe the ultrastructure of the nephron. In addition, stereology was used to quantify the volumes of cortex, medulla, and major vasculature of the kidneys, and the volumes and surface areas occupied by individual nephron components. There appeared to be no differences in the ultrastructural anatomy of the nephrons among the sparrows. Proximal tubules contained both narrow and wide intercellular spaces filled with interdigitations of the basolateral membrane. The thin limbs of Henle contained very wide intercellular spaces which were absent in the thick limbs of Henle. The distal tubule cells contained short, apical microvilli and infoldings of the basolateral membrane. In cross section, the medullary cones of all birds display an outer ring of thick limbs of Henle which surround an inner ring of collecting ducts, which in turn surround a central core of thin limbs of Henle. The Savannah Sparrow has a significantly higher volume of medulla compared to the two more mesic species. Within the cortex, the Savannah Sparrow also has a significantly higher volume of proximal tubules but a significantly lower volume of distal tubules than the other species. Within the medulla, the Savannah Sparrow has a significantly higher volume and surface area of capillaries, and a significantly higher surface area of thick limbs of Henle and collecting ducts than the mesic species. These data suggest that the salt marsh Savannah Sparrow has the renal morphology necessary to produce a more highly concentrated urine than the mesic zone species. PMID- 10681474 TI - Location of the vector of jaw muscle force in mammals. AB - Previous work has suggested that the third molar lies just in front of the point where the resultant vector of jaw muscle force, estimated from dissections, intersects the tooth row. This point meets the jaw such that the vector is 30% of jaw length from the jaw joint. Thus, the vector divides the jaw in the ratio of 3:7 when measurements are taken perpendicular to the vector. In practice, however, distances along mammalian jaws are typically measured on an easily determined line such as a line from one end of the tooth row to the other. The position of the jaw joint is then projected onto this line. As a rule, such a line is not perpendicular to the vector and so the distance from the projection of the joint, out to the rear of the third molar (and the vector's intersection), is different in different mammals. Rarely is this distance 30% of total jaw length. However, when the location of the vector's intersection is measured along the tooth row, this position varies directly with the inclination of the vector; a vector inclined posteriorly intersects the tooth row far from the projection of the joint and an anterior vector's intersection is relatively close. Only a vector perpendicular to the line from one end of the tooth row to the other intersects at 30%. This obvious point suggests a way to test the above hypotheses when the inclination of the vector is not known exactly. The predicted relationship between the distance to the molar, as a percentage of the total jaw length, and the approximate inclination of the vector derived from muscle weights (posterior or anterior depending on whether the temporalis or the masseter/pterygoid, respectively, is dominant) was observed in a sample of 46 different mammals. PMID- 10681476 TI - Are allografts the 'choice' in infectious endocarditis with periannular abscess? PMID- 10681477 TI - Functional assessment of coronary stenosis: it does make sense, but why don't I do it more often? PMID- 10681478 TI - Neural-natriuretic hormone interactions. PMID- 10681479 TI - Depression: the sleeping giant. PMID- 10681480 TI - May aspirin be replaced in the treatment of myocardial infarction? PMID- 10681481 TI - QT dispersion in ischaemic heart disease. PMID- 10681482 TI - Why are smoking cessation strategies not implemented more effectively in clinical practice? PMID- 10681483 TI - Heart rate variability: a telltale of health or disease. PMID- 10681484 TI - Diagnosis and treatment of nicotine dependence with emphasis on nicotine replacement therapy. A status report. AB - Tobacco use is a global health care problem. Repetitive exposure to nicotine produces neuroadaptation resulting in nicotine dependence. Cigarette smoking is particularly addictive due to the repeated delivery of bolus doses of nicotine to the bloodstream. Although compulsive tobacco use is sustained by nicotine addiction, it is the toxic combustion products in tobacco smoke such as carbon monoxide and oxidant gases that adversely affect the cardiovascular system. Smoking cessation produces significant health benefits and is a very cost effective intervention. Evidence that nicotine is the addictive component of tobacco provides the rationale for using nicotine replacement therapy to aid cessation. Nicotine replacement therapy doubles successful smoking cessation rates and evidence-based guidelines for the treatment of tobacco addiction recommend routine use of nicotine replacement therapy, particularly in heavily dependent smokers. Success rates of up to 40% can be achieved in specialist clinics. Despite early concerns regarding the safety of nicotine replacement therapy in smokers with heart disease, it is now clear that the health risks of using nicotine replacement therapy to assist such patients to stop, or significantly reduce, smoking far outweigh any treatment-related risks. PMID- 10681485 TI - Dispersion of ventricular repolarization is determined by the presence of myocardial viability in patients with old myocardial infarction. A dobutamine stress echocardiography study. AB - AIMS: The study sought to investigate the relationship of myocardial viability detected by dobutamine stress echocardiography to changes of QT dispersion and to the presence of arrhythmias during dobutamine infusion in patients with old myocardial infarction. We also examined whether patency of the infarct-related artery is associated with the presence of myocardial viability and QT dispersion. BACKGROUND: QT dispersion and myocardial variability have been associated with the presence of arrhythmias during late post infarction but not during dobutamine stress. Restoration of anterograde coronary flow has beneficial effects on ventricular systolic function and repolarization, suggesting that the extent of viable myocardium may determine ventricular repolarization. METHODS: Seventy five patients with previous myocardial infarction were studied in a low dose (up to 20 microg(-1) x kg(-1) x min(-1)) dobutamine stress echocardiography study. ECGs were obtained at rest and peak stress for measurement of QT intervals. The presence of ventricular arrhythmias (Lown grade >lb) during stress was noted. A reduction in the total wall motion score of the left ventricle at peak stress confirmed the presence of myocardial viability. RESULTS: Dobutamine infusion increased QT dispersion in all patients (P<0.01). Patients with myocardial viability had a lower resting QT dispersion (P<0.05) and a greater increase in QT dispersion% (P<0.01) than patients without. The combination of a resting QT dispersion <65 ms or an increase in QT dispersion >30% predicted viability with a sensitivity of 67%, a specificity of 96%, and an accuracy of 78%. A patent infarct-related artery, as well as ventricular arrhythmias, were more commonly observed in patients with evidence of viable myocardium (P<0.05). Patients with arrhythmias had a higher QT dispersion than patients without (P<0.05). CONCLUSION: The combination of a resting QT dispersion +/-65 ms or an increase in QT dispersion >30% predicts the presence of viable myocardium and thus, may represent a simple index for the assessment of viability in everyday clinical practice. Myocardial viability is related to a patent coronary artery and to a high incidence of arrhythmias accompanied by a greater increase in QT dispersion at peak dobutamine infusion. PMID- 10681486 TI - Randomized comparative trial of triflusal and aspirin following acute myocardial infarction. AB - AIMS: To compare the efficacy and tolerability of the antiplatelet agent triflusal with aspirin in the prevention of cardiovascular events following acute myocardial infarction. METHODS AND RESULTS: In this double-blind, multicentre, sequential design study, patients were randomized within 24 h of acute myocardial infarction symptom onset to receive triflusal 600 mg or aspirin 300 mg once daily for 35 days. The primary end-point was death, non-fatal myocardial reinfarction or a non-fatal cerebrovascular event. The incidences of these individual outcomes and urgent revascularization were secondary end-points. The null hypothesis of no difference between treatments in the primary combined end-point was accepted with 80% power after recruiting 2124 validated patients (odds ratio (OR) for failure [95% confidence interval (CI)]: 0.882 [0.634-1.227]). Non-fatal cerebrovascular events were significantly less frequent with triflusal (OR [95% CI]: 0.364 [0.146 0.908]; P = 0.030). There was no significant difference between treatments for death (OR [95% CI]: 0.816 [0.564-1.179]; P = 0.278), non-fatal reinfarction (OR [95% CI]: 1.577 [0.873-2.848]; P = 0.131) or revascularization (OR [95% CI]: 0.864 [0.644-1.161]; P = 0.334). Overall, both drugs were well tolerated, although there was a trend towards fewer bleeding episodes with triflusal; significantly fewer central nervous system bleeding episodes were observed in triflusal-treated patients (0.27% vs. 0.97%; P = 0.033). CONCLUSION: Triflusal and aspirin have similar efficacy in preventing further cardiovascular events after acute myocardial infarction, but triflusal showed a more favourable safety profile. Triflusal significantly reduced the incidence of non-fatal cerebrovascular events compared with aspirin. PMID- 10681487 TI - Angiographical and Doppler flow-derived parameters for assessment of coronary lesion severity and its relation to the result of exercise electrocardiography. DEBATE study group. Doppler Endpoints Balloon Angioplasty Trial Europe. AB - AIMS: Evaluation of angiographical and intracoronary Doppler-derived parameters of coronary stenosis severity. METHODS AND RESULTS: A total of 225 patients with one-vessel disease were studied before PTCA and at 6 months follow-up. Exercise electrocardiography was performed to document presence (n = 157) or absence (n = 138) of an ST segment shift (> or =0.1 mV). Intracoronary blood flow velocity analysis was performed to determine the proximal/distal flow velocity ratio, the distal diastolic/systolic flow velocity ratio and coronary flow velocity reserve. Receiver operator characteristic curves were calculated to assess the predictive value of these variables compared with the exercise test. The distal coronary flow velocity reserve demonstrated the best linear correlation for both percentage diameter stenosis and minimum lumen diameter (r = 0.67 and r = 0.66; P<0.01), compared to the diastolic/systolic flow velocity ratio (r = 0.19 and r = 0.14; P<0.01) and the proximal/distal flow velocity ratio (r = 0.03 and r = 0.07; not significant). The areas under the curve were 0. 84+/-0.02; 0.82+/-0.03 and 0.83+/-0.03 for diameter stenosis, minimum lumen diameter and coronary flow velocity reserve, respectively. Logistic regression analysis revealed that the percentage diameter stenosis or minimum lumen diameter and coronary flow velocity reserve were independent predictors for the result of stress testing. CONCLUSIONS: The distal coronary flow velocity reserve is the best intracoronary Doppler parameter for evaluation of coronary narrowings. Angiographical estimates of coronary lesion severity and distal coronary flow velocity reserve are good and independent predictors for the assessment of functional severity of coronary stenosis, emphasizing the complementary role of these parameters for clinical decision making. PMID- 10681488 TI - Depressed low frequency power of heart rate variability as an independent predictor of sudden death in chronic heart failure. AB - AIMS: Identification of patients with chronic heart failure at risk for sudden death remains difficult. We sought to assess the prognostic value for all-cause and sudden death of time and frequency domain measures of heart rate variability in chronic heart failure. METHODS AND RESULTS: We prospectively enrolled 190 patients with chronic heart failure in sinus rhythm, mean age 61+/-12 years, 109 (57.4%) in NYHA class II and 81 (42.6%) in classes III or IV, mean cardiothoracic ratio 57.6+/-6.4% and mean left ventricular ejection fraction 28.2+/-8.8%, 85 (45%) with ischaemic and 105 (55%) with idiopathic dilated cardiomyopathy. Time and frequency domain measures of heart rate variability were obtained from 24 h Holter ECG recordings, spectral measures were averaged for calculation of daytime (1000h-1900h) and night-time (2300h-0600h) values. During follow-up (22+/-18 months), 55 patients died, 21 of them suddenly and two presented with a syncopal spontaneous sustained ventricular tachycardia. In multivariate analysis, independent predictors for all-cause mortality were: ischaemic heart disease, cardiothoracic ratio > or =60% and standard deviation of all normal RR intervals <67 ms (RR = 2.5, 95% CI 1.5-4.2). Independent predictors of sudden death were: ischaemic heart disease and daytime low frequency power <3.3 ln (ms(2)) (RR = 2.8, 95% CI 1.2-8.6). CONCLUSION: Depressed heart rate variability has independent prognostic value in patients with chronic heart failure; spectral analysis identifies an increased risk for sudden death in these patients. PMID- 10681489 TI - Atrial right-to-left shunting causing severe hypoxaemia despite normal right sided pressures. Report of 11 consecutive cases corrected by percutaneous closure. AB - BACKGROUND: Hypoxaemia resulting from a right-to-left shunt occurs in patients with atrial septal defects and high pulmonary vascular resistance, but it is uncommon without pulmonary hypertension. METHODS: We report on 11 consecutive patients (age: 59-78 years) in whom a patent foramen ovale or a small atrial septal defect with normal right-sided pressures led to significant cyanosis with clinical symptoms. Six of them had associated platypnoea and orthodeoxia. The diagnosis was confirmed by contrast transoesophageal echocardiography showing an atrial right-to-left shunt. RESULTS: All but one were successfully treated by percutaneous closure of the inter-atrial defect. In one patient, delivery of the occluder failed due to kinking of the introducing sheath. Four complications were observed following the procedure: two supraventricular arrhythmias and a cerebrovascular accident, all resolved without sequelae; one patient died from a septic shock unrelated to the procedure. During follow-up (up to 30 months), no patient experienced any episode of desaturation due to inter-atrial shunting. CONCLUSION: Cyanosis without pulmonary arterial hypertension in the adult should prompt the performance of contrast transoesophageal echocardiography to identify a possible atrial right-to-left shunt. Percutaneous closure of the defect allows efficient and rapid correction of the hypoxaemia and avoids the need for surgical closure. PMID- 10681490 TI - Surgical treatment of active infective aortic valve endocarditis with associated periannular abscess--11 year results. AB - AIMS: The aim of the study was to evaluate the long-term results of allograft and prosthetic valve replacement in the treatment of infective aortic valve endocarditis with periannular abscess. METHODS: Between March 1988 and March 1996, 65 patients underwent surgery for active aortic valve endocarditis and paravalvular abscess. The indications for surgery were congestive heart failure, systemic emboli and atrioventricular block III. The pre-operative evaluation was performed with transoesophageal echocardiography. Aortic valve replacement was performed with allografts in 47 cases, with mechanical valves in 15, and bioprosthetic valves in three cases. All patients with total ventricular-aortic dehiscence and prosthetic valve endocarditis were treated with allografts. RESULTS: The 30-day mortality rate was 23.5% in the prosthetic group, when compared with 8.5% in the patients treated with allografts. The rate of recurrent valve infections during the 11-year follow-up period was 27.1% in the prosthetic group and 3.2% in the allograft group. The actuarial 11-year survival rate was 82.1% in the allograft group and 64.7% in the prosthetic group. CONCLUSION: Aortic allografts are an effective treatment for infective aortic valve endocarditis with associated periannular abscess. The operative mortality and recurrent infection rates are lower than in the prosthetic group, resulting in a significantly higher survival rate. Diagnosis and surgical management of these cases should be based on pre-operative transoesophageal echocardiography. PMID- 10681491 TI - Is the secretion of atrial natriuretic peptide in man under neural control? AB - AIMS: Previous work has described short-term variation in the circulating plasma level of atrial natriuretic peptide (ANP), but the mechanism remains unknown. Our aim was to investigate the role of cardiac innervation in this variability. METHODS AND RESULTS: Blood samples were obtained from the right atrium via a pulmonary artery flotation catheter every 2 min over a 90 min period. Seven patients who underwent cardiac transplantation by the standard biatrial technique (partial innervation) and ten patients who underwent transplantation by the bicaval technique (total denervation) were studied. ANP levels were measured by radioimmunoassay. The median ANP levels were somewhat higher in the biatrial group compared to the bicaval group [470 (150-1095) vs. 216 (100-605) pg. ml(-1); median (range); P = ns], and both were much higher than normal levels in the pulmonary artery (40 (24, 56) pg ml(-1); median and interquartile range). In both transplant groups circulating plasma ANP levels showed considerable variability. The median number of 'peaks' and 'troughs', as counted by visual inspection, were not significantly different between the two groups. Computer analysis identified 12-16 and 6-15 'pulses' in the biatrial and bicaval group, respectively. Further analysis revealed that pulse amplitude, height and area were significantly higher in the biatrial compared to the bicaval group. CONCLUSION: It would appear that variability of circulating plasma levels of ANP is preserved despite complete or partial cardiac denervation, and so a neural mechanism does not appear to account for such variation. PMID- 10681493 TI - ESC news and appointments PMID- 10681492 TI - ESC note from the publisher and the editor PMID- 10681494 TI - Syndecan-4 deficiency impairs focal adhesion formation only under restricted conditions. AB - Two domains of fibronectin deliver two different but cooperative signals required for focal adhesion formation. The signal from the cell-binding domain is mediated by integrins, whereas the signal from the heparin-binding domain is recognized by heparan sulfate proteoglycans, of which syndecan-4 has been hypothesized to be involved in focal adhesion formation. We generated mice deficient in syndecan-4 to study its role directly. Even in fibroblasts from syndecan-4-deficient mice, focal adhesions were formed, and actin fibers terminated normally at focal adhesions when they were cultured on coverslips coated with fibronectin or with a mixture of its cell-binding and heparin-binding fragments. However, when the cells were cultured on the cell-binding fragment and the heparin-binding fragment was added to the medium, focal adhesion formation was impaired in the syndecan-4 null fibroblasts as compared with that in wild-type cells. Therefore, syndecan-4 is essential for promoting focal adhesion formation only when the signal of the heparin-binding domain of fibronectin is delivered as a soluble form, most probably from the apical surface. When the signal is delivered as a substratum bound form, other molecule(s) also participate(s) in the signal reception. PMID- 10681495 TI - The packing of the transmembrane segments of human multidrug resistance P glycoprotein is revealed by disulfide cross-linking analysis. AB - Residues from several transmembrane (TM) segments of P-glycoprotein (P-gp) likely form the drug-binding site(s). To determine the organization of the TM segments, pairs of cysteine residues were introduced into the predicted TM segments of a Cys-less P-gp, and the mutant protein was subjected to oxidative cross-linking. In SDS gels, the cross-linked product migrated with a slower mobility than the native protein. The cross-linked products were not detected in the presence of dithiothreitol. Cross-linking was observed in 12 of 125 mutants. The pattern of cross-linking suggested that TM6 is close to TMs 10, 11, and 12, while TM12 is close to TMs 4, 5, and 6. In some mutants the presence of drug substrate colchicine, verapamil, cyclosporin A, or vinblastine either enhanced or inhibited cross-linking. Cross-linking was inhibited in the presence of ATP plus vanadate. These results suggest that the TM segments critical for drug binding must be close to each other and exhibit different conformational changes in response to binding of drug substrate or vanadate trapping of nucleotide. Based on these results, we propose a model for the arrangement of the TM segments. PMID- 10681496 TI - WD40 repeat proteins striatin and S/G(2) nuclear autoantigen are members of a novel family of calmodulin-binding proteins that associate with protein phosphatase 2A. AB - Protein phosphatase 2A (PP2A) is a multifunctional serine/threonine phosphatase that is critical to many cellular processes including development, neuronal signaling, cell cycle regulation, and viral transformation. PP2A has been implicated in Ca(2+)-dependent signaling pathways, but how PP2A is targeted to these pathways is not understood. We have identified two calmodulin (CaM)-binding proteins that form stable complexes with the PP2A A/C heterodimer and may represent a novel family of PP2A B-type subunits. These two proteins, striatin and S/G(2) nuclear autoantigen (SG2NA), are highly related WD40 repeat proteins of previously unknown function and distinct subcellular localizations. Striatin has been reported to associate with the post-synaptic densities of neurons, whereas SG2NA has been reported to be a nuclear protein expressed primarily during the S and G(2) phases of the cell cycle. We show that SG2NA, like striatin, binds to CaM in a Ca(2+)-dependent manner. In addition to CaM and PP2A, several unidentified proteins stably associate with the striatin-PP2A and SG2NA PP2A complexes. Thus, one mechanism of targeting and organizing PP2A with components of Ca(2+)-dependent signaling pathways may be through the molecular scaffolding proteins striatin and SG2NA. PMID- 10681497 TI - An essential glutamyl residue in EmrE, a multidrug antiporter from Escherichia coli. AB - EmrE is an Escherichia coli 12-kDa protein that confers resistance to toxic compounds, by actively removing them in exchange with protons. The protein includes eight charged residues. Seven of these residues are located in the hydrophilic loops and can be replaced with either Cys or another amino acid bearing the same charge, without impairing transport activity. Glu-14 is the only charged residue in the membrane domain and is conserved in all the proteins of the family. We show here that this residue is the site of action of dicyclohexylcarbodiimide, a carbodiimide known to act in hydrophobic environments. When Glu-14 was replaced with either Cys or Asp, resistance was abolished. Whereas the E14C mutant displays no transport activity, the E14D protein shows efflux and exchange at rates about 30-50% that of the wild type. The maximal DeltapH-driven uptake rate of E14D is only 10% that of the wild type. The mutant shows a different pH profile in all the transport modes. Our results support the notion that Glu-14 is an essential part of a binding domain shared by substrates and protons but mutually exclusive in time. This notion provides the molecular basis for the obligatory exchange catalyzed by EmrE. PMID- 10681498 TI - Proximity of periplasmic loops in the metal-Tetracycline/H(+) antiporter of Escherichia coli observed on site-directed chemical cross-linking. AB - Our previous study on second-site suppressor mutations of the Tn10-encoded metal tetracycline/H(+) antiporter suggested that Leu(30) and Ala(354), located in periplasmic loop 1-2 and 11-12, respectively, are conformationally linked to each other (Kawabe, T., and Yamaguchi, A. (1999) FEBS Lett. 457, 169-173). To determine the spatial proximity of these two residues, cross-linking gel-shift assays of the L30C/A354C double mutant were performed after the mutant had been oxidized with Cu(2+)/o-phenanthroline. The results indicated that Leu(30) and Ala(354) are close to each other but that Gly(62), which is located in cytoplasmic loop 2-3, and Ala(354) are distant from each other, as a negative control. Then, a single Cys residue was introduced into each of the six periplasmic loop regions (P1-P6), and eleven double mutants were constructed. Of these eleven double Cys mutants, the L30C/A354C and L30C/T235C mutants showed a mobility shift on oxidation, indicating that P1 is spatially close to P4 as well as P6. In contrast, the other nine mutants, L30C/S92C, L30C/S156C, L30C/S296C, S92C/S296C, S92C/T235C, S92C/A354C, S156C/T235C, S156C/S296C, and S156C/A354C, showed no mobility shift under oxidized conditions on intramolecular cross linking. The S92C and S296C mutants showed dimerization on intermolecular cross linking, indicating that P2 and P5 are located at the periphery of the helix bundle. PMID- 10681499 TI - On the spatial organization of hemes and chlorophyll in cytochrome b(6)f. A linear and circular dichroism study. AB - The organization of chromophores in the cytochrome b(6) f from Chlamydomonas reinhardtii has been studied spectroscopically. Linear dichroism (LD) measurements, performed on the complex co-reconstituted into vesicles with photosynthetic reaction centers as an internal standard, allow the determination of the orientations of the chromophore with respect to the membrane plane. The orientations of the b(H)- and b(L)-hemes are comparable to those determined crystallographically on the cytochrome bc(1). The excitonic CD signal, resulting from the interaction between b-hemes, is similar to that reported for the cytochrome bc(1). LD and CD data are consistent with the differences between the b(6) f and bc(1) leaving the orientation of the b-hemes unaffected. By contrast, the LD data yield a different orientation for the heme f as compared either to the heme c(1) in the crystallographic structures or to the heme f as studied by electron paramagnetic resonance. This difference could either result from incorrect assumptions regarding the orientations of the electronic transitions of the f-heme or may point to the possibility of a redox-dependent movement of cytochrome f. The chlorophyll a was observed in a well defined orientation, further corroborating a specific binding site for it in the b(6) f complex. PMID- 10681500 TI - Relative orientation of the hemes of the cytochrome bc(1) complexes from Rhodobacter sphaeroides, Rhodospirillum rubrum, and beef heart mitochondria. A linear dichroism study. AB - The orientation of the chromophores in the cytochrome bc(1) of Rhodospirillum rubrum, Rhodobacter sphaeroides, and beef heart mitochondria is reported. The combination of redox-resolved absorption spectrophotometry and linear dichroism experiments at low temperature allows the determination of the orientation of the three hemes with respect to the membrane plane. The orientations of the b(H)-and b(L)-hemes of the R. sphaeroides and beef heart mitochondrial complexes are similar to those determined by crystallographic studies of the mitochondrial cytochrome bc(1). On the other hand the orientations of the b-hemes of the R. rubrum complex lead to the conclusion that the b(H)-heme is more perpendicular to the membrane plane than the b(L)-heme. This could be explained by a specific organization of the b-hemes due to subunit composition of the complex or, alternatively, to a different spatial position of the heme transitions with respect to the porphyrin macrocycle compared with the other complexes. Moreover, our results demonstrate a different orientation of the heme c(1) of the three studied complexes in comparison to crystallographic studies. This difference may arise from the above hypothesis on the transitions of the heme or from flexibility of this subunit in function of its redox state. PMID- 10681501 TI - Interaction of endothelial and neuronal nitric-oxide synthases with the bradykinin B2 receptor. Binding of an inhibitory peptide to the oxygenase domain blocks uncoupled NADPH oxidation. AB - Endothelial nitric-oxide synthase (type III) (eNOS) was reported to form an inhibitory complex with the bradykinin receptor B2 (B2R) from which the enzyme is released in an active form upon receptor activation (Ju, H., Venema, V. J., Marrero, M. B., and Venema, R. C. (1998) J. Biol. Chem. 273, 24025-24029). Using a synthetic peptide derived from the known inhibitory sequence of the B2R (residues 310-329) we studied the interaction of the receptor with purified eNOS and neuronal nitric-oxide synthase (type I) (nNOS). The peptide inhibited formation of L-citrulline by eNOS and nNOS with IC(50) values of 10.6 +/- 0.4 microM and 7.1 +/- 0.6 microM, respectively. Inhibition was not due to an interference of the peptide with L-arginine or tetrahydrobiopterin binding. The NADPH oxidase activity of nNOS measured in the absence of L-arginine was inhibited by the peptide with an IC(50) of 3.7 +/- 0.6 microM, but the cytochrome c reductase activity of the enzyme was much less susceptible to inhibition (IC(50) >0.1 mM). Steady-state absorbance spectra of nNOS recorded during uncoupled NADPH oxidation showed that the heme remained oxidized in the presence of the synthetic peptide consisting of amino acids 310-329 of the B2R, whereas the reduced oxyferrous heme complex was accumulated in its absence. These data suggest that binding of the B2R 310-329 peptide blocks flavin to heme electron transfer. Co-immunoprecipitation of B2R and nNOS from human embryonic kidney cells stably transfected with human nNOS suggests that the B2R may functionally interact with nNOS in vivo. This interaction of nNOS with the B2R may recruit the enzyme to allow for the effective coupling of bradykinin signaling to the nitric oxide pathway. PMID- 10681502 TI - Reaction intermediates and single turnover rate constants for the oxidation of heme by human heme oxygenase-1. AB - Heme oxygenase converts heme to biliverdin, iron, and CO in a reaction with two established intermediates, alpha-meso-hydroxyheme and verdoheme. Transient kinetic studies show that the conversion of Fe(3+)-heme to Fe(3+)-verdoheme is biphasic. Electron transfer to the heme (0.11 s(-1) at 4 degrees C and 0.49 s(-1) at 25 degrees C) followed by rapid O(2) binding yields the ferrous dioxy complex. Transfer of an electron (0.056 s(-1) at 4 degrees C and 0.21 s(-1) at 25 degrees C) to this complex triggers the formation of alpha-meso-hydroxyheme and its subsequent O(2)-dependent fragmentation to Fe(3+)-verdoheme. The conversion of Fe(3+)-verdoheme to Fe(3+)-biliverdin is also biphasic. Thus, reduction of Fe(3+) to Fe(2+)-verdoheme (0.15 s(-1) at 4 degrees C and 0.55 s(-1) at 25 degrees C) followed by O(2) binding and an electron transfer produces Fe(3+)-biliverdin (0.025 s(-1) at 4 degrees C and 0.10 s(-1) at 25 degrees C). The conversion of Fe(3+)-biliverdin to free biliverdin is triphasic. Reduction of Fe(3+)-biliverdin (0.035 s(-1) at 4 degrees C and 0.15 s(-1) at 25 degrees C), followed by rapid release of Fe(2+) (0.19 s(-1) at 4 degrees C and 0.39 s(-1) at 25 degrees C), yields the biliverdin-enzyme complex from which biliverdin slowly dissociates (0.007 s(-1) at 4 degrees C and 0.03 s(-1) at 25 degrees C). The rate of Fe(2+) release agrees with the rate of Fe(3+)-biliverdin reduction. Fe(2+) release clearly precedes biliverdin dissociation. In the absence of biliverdin reductase, biliverdin release is the rate-limiting step, but in its presence biliverdin release is accelerated and the overall rate of heme degradation is limited by the conversion of Fe(2+)-verdoheme to the Fe(3+)-biliverdin. PMID- 10681503 TI - Cloning and characterization of RAP250, a novel nuclear receptor coactivator. AB - Ligand-induced transcriptional activation of gene expression by nuclear receptors is dependent on recruitment of coactivators as intermediary factors. The present work describes the cloning and characterization of RAP250, a novel human nuclear receptor coactivator. The results of in vitro and in vivo experiments indicate that the interaction of RAP250 with nuclear receptors is ligand-dependent or ligand-enhanced depending on the nuclear receptor and involves only one short LXXLL motif called nuclear receptor box. Transient transfection assays further demonstrate that RAP250 has a large intrinsic glutamine-rich activation domain and can significantly enhance the transcriptional activity of several nuclear receptors, acting as a coactivator. Interestingly, Northern blot and in situ hybridization analyses reveal that RAP250 is widely expressed with the highest expression in reproductive organs (testis, prostate and ovary) and brain. Together, our data suggest that RAP250 may play an important role in mammalian gene expression mediated by nuclear receptor. PMID- 10681504 TI - The acidic triad conserved in type IA DNA topoisomerases is required for binding of Mg(II) and subsequent conformational change. AB - The acidic residues Asp-111, Asp-113, and Glu-115 of Escherichia coli DNA topoisomerase I are located near the active site Tyr-319 and are conserved in type IA topoisomerase sequences with counterparts in type IIA DNA topoisomerases. Their exact functional roles in catalysis have not been clearly defined. Mutant enzymes with two or more of these residues converted to alanines were found to have >90% loss of activity in the relaxation assay with 6 mM Mg(II) present. Mg(II) concentrations (15-20 mM) inhibitory for the wild type enzyme are needed by these double mutants for maximal relaxation activity. The triple mutant D111A/D113A/E115A had no detectable relaxation activity. Mg(II) binding to wild type enzyme resulted in an altered conformation detectable by Glu-C proteolytic digestion. This conformational change was not observed for the triple mutant or for the double mutant D111A/D113A. Direct measurement of Mg(II) bound showed the loss of 1-2 Mg(II) ions for each enzyme molecule due to the mutations. These results demonstrate a functional role for these acidic residues in the binding of Mg(II) to induce the conformational change required for the relaxation of supercoiled DNA by the enzyme. PMID- 10681505 TI - 5'-nicked apurinic/apyrimidinic sites are resistant to beta-elimination by beta polymerase and are persistent in human cultured cells after oxidative stress. AB - Genomic DNA is continuously exposed to oxidative stress. Whereas reactive oxygen species (ROS) preferentially react with bases in DNA, free radicals also abstract hydrogen atoms from deoxyribose, resulting in the formation of apurinic/apyrimidinic (AP) sites and strand breaks. We recently reported high steady-state levels of AP sites in rat tissues and human liver DNA (Nakamura, J., and Swenberg, J. A. (1999) Cancer Res. 59, 2522-2526). These AP sites were predominantly cleaved 5' to the lesion. We hypothesized that these endogenous AP sites were derived from oxidative stress. In this investigation, AP sites induced by ROS were quantitated and characterized. A combination of H(2)O(2) and FeSO(4) induced significant numbers of AP sites in calf thymus DNA, which were predominantly cleaved 5' to the AP sites (75% of total aldehydic AP sites). An increase in the number of 5'-AP sites was also detected in human cultured cells exposed to H(2)O(2), and these 5'-AP sites were persistent during the post exposure period. beta-Elimination by DNA beta-polymerase efficiently excised 5' regular AP sites, but not 5'-AP sites, in DNA from cells exposed to H(2)O(2). These results suggest that 5'-oxidized AP sites induced by ROS are not efficiently repaired by the mammalian short patch base excision repair pathway. PMID- 10681506 TI - Poliovirus RNA-dependent RNA polymerase (3D(pol)). Assembly of stable, elongation competent complexes by using a symmetrical primer-template substrate (sym/sub). AB - Detailed studies of the kinetics and mechanism of nucleotide incorporation catalyzed by the RNA-dependent RNA polymerase from poliovirus, 3D(pol), have been limited by the inability to assemble elongation complexes that permit activity to be monitored by extension of end-labeled primers. We have solved this problem by employing a short, symmetrical, heteropolymeric RNA primer-template that we refer to as "sym/sub." Formation of 3D(pol)-sym/sub complexes is slow owing to a slow rate of association (0.1 microM(-1) s(-1)) of 3D(pol) and sym/sub and a slow isomerization (0. 076 s(-1)) of the 3D(pol)-sym/sub complex that is a prerequisite for catalytic competence of this complex. Complex assembly is stoichiometric under conditions in which competing reactions, such as enzyme inactivation, are eliminated. Inactivation of 3D(pol) occurs at a maximal rate of 0.051 s(-1) at 22 degrees C in reaction buffer lacking nucleotide. At this temperature, ATP protects 3D(pol) against inactivation with a K(0.5) of 37 microM. Once formed, 3D(pol)-sym/sub elongation complexes are stable (t((1)/(2)) = 2 h at 22 degrees C) and appear to contain only a single polymerase monomer. In the presence of Mg(2+), AMP, 2'-dAMP, and 3'-dAMP are incorporated into sym/sub by 3D(pol) at rates of 72, 0.6, and 1 s(-1), respectively. After incorporation of AMP, 3D(pol)-sym/sub product complexes have a half-life of 8 h at 22 degrees C. The stability of 3D(pol)-sym/sub complexes is temperature-dependent. At 30 degrees C, there is a 2-8-fold decrease in complex stability. Complex dissociation is the rate-limiting step for primer utilization. 3D(pol) dissociates from the end of template at a rate 10-fold faster than from internal positions. The sym/sub system will facilitate mechanistic analysis of 3D(pol) and permit a direct kinetic and thermodynamic comparison of the RNA-dependent RNA polymerase to the other classes of nucleic acid polymerases. PMID- 10681507 TI - Kv4.2 phosphorylation by cyclic AMP-dependent protein kinase. AB - Recent evidence suggests that K(+) channels composed of Kv4.2 alpha-subunits underlie a transient current in hippocampal CA1 neurons and ventricular myocytes, and activation of the cAMP second messenger cascade has been shown to modulate this transient current. We determined if Kv4.2 alpha-subunits were directly phosphorylated by cAMP-dependent protein kinase (PKA). The intracellular domains of the amino and carboxyl termini of Kv4.2 were expressed as glutathione S transferase fusion protein constructs; we observed that both of these fusion proteins were substrates for PKA in vitro. By using phosphopeptide mapping and amino acid sequencing, we identified PKA phosphorylation sites on the amino- and carboxyl-terminal fusion proteins corresponding to Thr(38) and Ser(552), respectively, within the Kv4.2 sequence. Kinetic characterization of the PKA sites demonstrated phosphorylation kinetics comparable to Kemptide. To evaluate PKA site phosphorylation in situ, phospho-selective antisera for each of the sites were generated. By using COS-7 cells expressing an EGFP-Kv4.2 fusion protein, we observed that stimulation of the endogenous PKA cascade resulted in an increase in phosphorylation of Thr(38) and Ser(552) within Kv4.2 in the intact cell. We also observed modulation of PKA phosphorylation at these sites within Kv4.2 in hippocampal area CA1. These results provide insight into likely sites of regulation of Kv4.2 by PKA. PMID- 10681508 TI - TA1/LAT-1/CD98 light chain and system L activity, but not 4F2/CD98 heavy chain, respond to arginine availability in rat hepatic cells. Loss Of response in tumor cells. AB - Tumor associated gene-1/L amino acid transporter-1 (TA1/LAT-1) was recently identified as a light chain of the CD98 amino acid transporter and cellular activation marker. Our previous studies with primary rat hepatocyte cultures demonstrated that TA1 RNA levels were responsive to media amino acid concentrations, suggesting adaptive regulation. High level TA1 expression associated with transformed cells also suggested a role in tumor progression. The present study examined the relationship of TA1/CD98 expression, adaptive response, and associated amino acid transport to neoplastic transformation using a panel of well characterized rat hepatic cell lines. We found 1) increased expression of TA1 in response to amino acid depletion, specific for arginine but not glutamine; 2) loss of TA1 response to arginine in gamma-glutamyl transpeptidase-positive transformed and tumorigenic cells; 3) no appreciable response of 4F2/CD98 heavy chain to arginine levels; and 4) correlation of system L amino acid transport activity in response to arginine with changes in TA1/LAT-1 mRNA but not total immunoreacting protein. Our results suggest this CD98 light chain may act as an environmental sensor, responding to amino acid availability and that its regulation is complex. We hypothesize that altered TA1 expression is an early event in hepatocarcinogenesis giving neoplastic cells a growth or survival advantage, particularly under conditions of limited amino acid availability. PMID- 10681509 TI - Effect of ethanol and osmotic stress on receptor conformation. Reduced water activity amplifies the effect of ethanol on metarhodopsin II formation. AB - The combined effects of ethanol and osmolytes on both the extent of formation of metarhodopsin II (MII), which binds and activates transducin, and on acyl chain packing were examined in rod outer segment disc membranes. The ethanol-induced increase in MII formation was amplified by the addition of neutral osmolytes. This enhancement was linear with osmolality. At 360 milliosmolal, the osmolality of human plasma, 50 mM ethanol was 2.7 times more potent than at 0 osmolality, demonstrating the importance of water activity in in vitro experiments dealing with ethanol potency. Ethanol disordered acyl chain packing, and increasing osmolality enhanced this acyl chain disordering. Prior osmotic stress data showed a release of 35 +/- 2 water molecules upon MII formation. Ethanol increases this number to 49 water molecules, suggesting that ethanol replaces 15 additional water molecules upon MII formation. Amplification of ethanol effects on MII formation and acyl chain packing by osmolytes suggests that ethanol increases the equilibrium concentration of MII both by disordering acyl chain packing and by disrupting rhodopsin-water hydrogen bonds, demonstrating a direct effect of ethanol on rhodopsin. At physiologically relevant levels of osmolality and ethanol, about 90% of ethanol's effect is due to disordered acyl chain packing. PMID- 10681510 TI - Phosphoinositide 3-kinase is involved in the tumor-specific activation of human breast cancer cell Na(+)/H(+) exchange, motility, and invasion induced by serum deprivation. AB - Whereas the tumor acidic extracellular pH plays a crucial role in the invasive process, the mechanism(s) behind this acidification, especially in low nutrient conditions, are unclear. The regulation of the Na(+)/H(+) exchanger (NHE) and invasion by serum deprivation were studied in a series of breast epithelial cell lines representing progression from non-tumor to highly metastatic cells. Whereas serum deprivation reduced lactate production in all three cells lines, it inhibited NHE activity in the non-tumor cells and stimulated it in the tumor cells with a larger stimulation in the metastatic cells. The stimulation of NHE in the tumor cell lines was the result of an increased affinity of the internal H(+) regulatory site of the NHE without changes in sodium kinetics or expression. Serum deprivation conferred increased cell motility and invasive ability that were abrogated by specific inhibition of the NHE. Inhibition of phosphoinositide 3-kinase by overexpression of a dominant-negative mutant or wortmannin incubation inhibited NHE activity and invasion in serum replete conditions while potentiating the serum deprivation-dependent activation of the NHE and invasion. These results indicate that the up-regulation of the NHE by a phosphoinositide 3 kinase-dependent mechanism plays an essential role in increased tumor cell invasion induced by serum deprivation. PMID- 10681511 TI - Disulfide-mediated oligomerization of Peripherin/Rds and Rom-1 in photoreceptor disk membranes. Implications for photoreceptor outer segment morphogenesis and degeneration. AB - Peripherin/Rds is a tetraspanning membrane protein that has been implicated in photoreceptor outer segment morphogenesis and inherited retinal degenerative diseases. Together with the structurally related protein, Rom-1, it forms a complex along the rims of rod and cone disc membranes. We have compared the oligomeric structure of these proteins from nonreduced and dithiothreitol reduced membranes by velocity sedimentation, SDS-gel electrophoresis, immunoaffinity chromatography, and chemical cross-linking. Under reducing conditions peripherin/Rds and Rom-1 existed as homomeric and heteromeric core complexes devoid of intermolecular disulfide bonds. Under nonreducing conditions core complexes associated through intermolecular disulfide bonds to form oligomers. One intermediate-size oligomer contained monomers and disulfide-linked dimers of peripherin/Rds and Rom-1, while larger oligomers consisted only of disulfide linked peripherin/Rds dimers when analyzed on nonreducing SDS gels. Consistent with this result, disc membranes contained twice as much peripherin/Rds as Rom-1. Peripherin/Rds individually expressed in COS-1 cells also formed disulfide-linked oligomers bridged through Cys-150 residues, whereas Rom-1 showed little tendency to form oligomers. These results indicate that peripherin/Rds and Rom-1 associate noncovalently to form multisubunit core complexes. Peripherin/Rds containing core complexes interact through specific intermolecular disulfide bonds to form oligomers which may play a crucial role in photoreceptor disc morphogenesis and retinal degenerative diseases. PMID- 10681512 TI - Ligand-, cell-, and estrogen receptor subtype (alpha/beta)-dependent activation at GC-rich (Sp1) promoter elements. AB - 17beta-Estradiol (E2) induces expression of several genes via estrogen receptor (ER)-Sp1 protein interactions with GC-rich promoter elements in which Sp1 but not ER binds DNA. This study reports the ligand- and cell context-dependent ER(alpha)/Sp1 and ER(beta)/Sp1 action using an E2-responsive construct (pSp1) containing a GC-rich promoter. Both ER(alpha) and ER(beta) proteins physically interact with Sp1 (coimmunoprecipitation) and preferentially bind to the C terminal region of this protein in pull-down assays. E2- and antiestrogen dependent transcriptional activation of ER(alpha)/Sp1 was observed in MCF-7, MDA MB-231, and LnCaP cells, but not in HeLa cells. E2 did not affect or significantly decrease ER(beta)/Sp1 action, and antiestrogens had minimal effects in the same 4 cell lines. Exchange of activation function-1 (AF-1) domains of ER subtypes gave chimeric ER(alpha/beta) (AF-1alpha/AF-2beta) and ER(beta/alpha) (AF 1beta/AF-2alpha) proteins that resembled wild-type ER (alpha or beta) in terms of physical association with Sp1 protein. Transcriptional activation studies with chimeric ER(beta/alpha) and ER(alpha/beta) showed that only ER(alpha/beta) can activate transcription from an Sp1 element, not ER(beta/alpha). This indicates that the AF-1 domain from ER(alpha) is responsible for activation at an Sp1 element, independent of ER subtype context. In order to further characterize this observation, deletion constructs in the AF-1 domain of both ER(alpha) and ER(alpha/beta) were made, and transactivation studies indicated that the region between amino acids 79 and 117 of this domain is important for activation at an Sp1 element. PMID- 10681513 TI - Nerve growth factor cooperates with p185(HER2) in activating growth of human breast carcinoma cells. AB - Nerve growth factor (NGF) is known to exert a mitogenic effect on human breast cancer cells through proto-TrkA activation. Reverse transcriptase-PCR analysis of proto-TrkA expression in human breast carcinoma specimens and cell lines revealed trkA transcript in 12 of 14 human breast carcinoma specimens and in all of four cell lines tested. While cytofluorimetric and Western blot analysis indicated proto-TrkA expression in three of the four cell lines, NGF stimulated growth in only two of the three positive cell lines. Inhibition of NGF-induced MAPK activation by an antibody directed against the extracellular domain of TrkA but not by an inhibitor of TrkA phosphorylation demonstrated the requirement of NGF binding but not of proto-TrkA kinase activity for MAPK activation, suggesting the recruitment of another kinase for transmission of the mitogenic signaling. Indeed, NGF induced tyrosine phosphorylation and stimulated kinase activity of p185(HER2), a kinase receptor of the HER family. A TrkA phosphorylation inhibitor did not affect this activation. Moreover, the two receptors were coprecipitated by antibodies directed against proto-TrkA and p185(HER2). Down-modulation of p185(HER2) expression in a breast carcinoma line transfected with a construct containing an anti-p185(HER2) antibody sequence and expressing proto-TrkA impaired NGF-induced MAPK activation and proliferation. Together these data show that in cells expressing low levels of TrkA such as breast carcinoma cells, NGF must recruit other overexpressed receptors such as p185(HER2) in order to generate a biological signal that can induce breast cancer cell growth. PMID- 10681514 TI - Overexpression of heme oxygenase in neuronal cells, the possible interaction with Tau. AB - Increased expression of heme oxygenase-1 (HO-1) is a common feature in a number of neurodegenerative diseases. Interestingly, the spatial distribution of HO-1 expression in diseased brain is essentially identical to that of pathological expression of tau. In this study, we explored the relationship between HO-1 and tau, using neuroblastoma cells stably transfected with sense and antisense HO-1 constructs as well as with the vector alone. In transfected cells overexpressing HO-1, the activity of heme oxygenase was increased, and conversely, the level of tau protein was dramatically decreased when compared with antisense HO-1 or CEP transfected cells. The suppression of tau protein expression was almost completely reversed by zinc-deuteroporphyrin, a specific inhibitor of heme oxygenase activity. The activated forms of ERKs (extracellular signal-regulated kinases) were also decreased in cells overexpressing HO-1 although no changes in the expression of total ERK-1/2 proteins were observed. These data are in agreement with the finding that the expression of tau is regulated through signal cascades including the ERKs, whose activities are modulated by oxidative stresses. The expression of tau and HO-1 may be regulated by oxidative stresses in a coordinated manner and play a pivotal role in the cytoprotection of neuronal cells. PMID- 10681515 TI - Fast kinetic analysis of conformational changes in mutants of the Ca(2+)-ATPase of sarcoplasmic reticulum. AB - Rapid quench experiments at 25 degrees C were carried out on selected mutants of the sarco(endo)plasmic reticulum Ca(2+)-ATPase to assess the kinetics of the conformational changes of the dephosphoenzyme associated with ATP binding/phosphoryl transfer and the binding and dissociation of Ca(2+) at the cytoplasmically facing transport sites. The mutants Gly(233) --> Glu, Gly(233) - > Val, Pro(312) --> Ala, Leu(319) --> Arg, and Lys(684) --> Arg differed conspicuously with respect to the behavior of the dephosphoenzyme, although they were previously shown to display a common block of the transformation of the phosphoenzyme from an ADP-sensitive to an ADP-insensitive form. The maximum rate of the ATP binding/phosphoryl transfer reaction was reduced 3.6-fold in mutant Gly(233) --> Glu and more than 50-fold in mutant Lys(684) --> Arg, relative to wild type. In mutant Leu(319) --> Arg, the rate of the Ca(2+)-binding transition was reduced as much as 10-30-fold depending on the presence of ATP. In mutants Gly(233) --> Glu, Gly(233) --> Val, and Pro(312) --> Ala, the rate of the Ca(2+) binding transition was increased at least 2-3-fold at acid pH but not significantly at neutral pH, suggesting a destabilization of the protonated form. The rate of Ca(2+) dissociation was reduced 12-fold in mutant Pro(312) --> Ala and 3.5-fold in Leu(319) --> Arg, and increased at least 4-fold in a mutant in which the putative Ca(2+) liganding residue Glu(309) was replaced by aspartate. The data support a model in which Pro(312) and Leu(319) are closely associated with the cation binding pocket, Gly(233) is part of a long-range signal transmission pathway between the ion-binding sites and the catalytic site, and Lys(684) is an essential catalytic residue that may function in the same way as its counterpart in the soluble hydrolases belonging to the haloacid dehalogenase superfamily. PMID- 10681516 TI - Mammalian 5'(3')-deoxyribonucleotidase, cDNA cloning, and overexpression of the enzyme in Escherichia coli and mammalian cells. AB - 5'(3')-Deoxyribonucleotidase is a ubiquitous enzyme in mammalian cells whose physiological function is not known. It was earlier purified to homogeneity from human placenta. We determined the amino acid sequences of several internal peptides and with their aid found an expressed sequence tag clone with the complete cDNA for a murine enzyme of 23.9 kDa. The DNA was cloned into appropriate plasmids and introduced into Escherichia coli and ecdyson-inducible 293 and V79 cells. The recombinant enzyme was purified to homogeneity from transformed E. coli and was found to be identical with the native enzyme. After induction with ponasterone, the transfected mammalian cells showed a gradual increase of enzyme activity. A human expressed sequence tag clone contained a large part of the cDNA of the human enzyme but lacked the 5'-end corresponding to 51 amino acids of the murine enzyme. Several polymerase chain reaction-based approaches to find this sequence met with no success. A mouse/human hybrid cDNA that had substituted the missing human 5'-end with the corresponding mouse sequence coded for a fully active enzyme. PMID- 10681517 TI - NAD(P)H:Quinone oxidoreductase activity is the principal determinant of beta lapachone cytotoxicity. AB - beta-Lapachone activates a novel apoptotic response in a number of cell lines. We demonstrate that the enzyme NAD(P)H:quinone oxidoreductase (NQO1) substantially enhances the toxicity of beta-lapachone. NQO1 expression directly correlated with sensitivity to a 4-h pulse of beta-lapachone in a panel of breast cancer cell lines, and the NQO1 inhibitor, dicoumarol, significantly protected NQO1 expressing cells from all aspects of beta-lapachone toxicity. Stable transfection of the NQO1-deficient cell line, MDA-MB-468, with an NQO1 expression plasmid increased apoptotic responses and lethality after beta-lapachone exposure. Dicoumarol blocked both the apoptotic responses and lethality. Biochemical studies suggest that reduction of beta-lapachone by NQO1 leads to a futile cycling between the quinone and hydroquinone forms, with a concomitant loss of reduced NAD(P)H. In addition, the activation of a cysteine protease, which has characteristics consistent with the neutral calcium-dependent protease, calpain, is observed after beta-lapachone treatment. This is the first definitive elucidation of an intracellular target for beta-lapachone in tumor cells. NQO1 could be exploited for gene therapy, radiotherapy, and/or chemopreventive interventions, since the enzyme is elevated in a number of tumor types (i.e. breast and lung) and during neoplastic transformation. PMID- 10681518 TI - Nitric oxide modulates the catalytic activity of myeloperoxidase. AB - Myeloperoxidase (MPO), an abundant protein in neutrophils, monocytes, and subpopulations of tissue macrophages, is believed to play a critical role in host defenses and inflammatory tissue injury. To perform these functions, an array of diffusible radicals and reactive oxidant species may be formed through oxidation reactions catalyzed at the heme center of the enzyme. Myeloperoxidase and inducible nitric-oxide synthase are both stored in and secreted from the primary granules of activated leukocytes, and nitric oxide (nitrogen monoxide; NO) reacts with the iron center of hemeproteins at near diffusion-controlled rates. We now demonstrate that NO modulates the catalytic activity of MPO through distinct mechanisms. NO binds to both ferric (Fe(III), the catalytically active species) and ferrous (Fe(II)) forms of MPO, generating stable low-spin six-coordinate complexes, MPO-Fe(III).NO and MPO-Fe(II).NO, respectively. These nitrosyl complexes were spectrally distinguishable by their Soret absorbance peak and visible spectra. Stopped-flow kinetic analyses indicated that NO binds reversibly to both Fe(III) and Fe(II) forms of MPO through simple one-step mechanisms. The association rate constant for NO binding to MPO-Fe(III) was comparable to that observed with other hemoproteins whose activities are thought to be modulated by NO in vivo. In stark contrast, the association rate constant for NO binding to the reduced form of MPO, MPO-Fe(II), was over an order of magnitude slower. Similarly, a 2-fold decrease was observed in the NO dissociation rate constant of the reduced versus native form of MPO. The lower NO association and dissociation rates observed suggest a remarkable conformational change that alters the affinity and accessibility of NO to the distal heme pocket of the enzyme following heme reduction. Incubation of NO with the active species of MPO (Fe(III) form) influenced peroxidase catalytic activity by dual mechanisms. Low levels of NO enhanced peroxidase activity through an effect on the rate-limiting step in catalysis, reduction of Compound II to the ground-state Fe(III) form. In contrast, higher levels of NO inhibited MPO catalysis through formation of the nitrosyl complex MPO-Fe(III)-NO. NO interaction with MPO may thus serve as a novel mechanism for modulating peroxidase catalytic activity, influencing the regulation of local inflammatory and infectious events in vivo. PMID- 10681519 TI - Loss of the major isoform of phosphoglucomutase results in altered calcium homeostasis in Saccharomyces cerevisiae. AB - Phosphoglucomutase (PGM) is a key enzyme in glucose metabolism, where it catalyzes the interconversion of glucose 1-phosphate (Glc-1-P) and glucose 6 phosphate (Glc-6-P). In this study, we make the novel observation that PGM is also involved in the regulation of cellular Ca(2+) homeostasis in Saccharomyces cerevisiae. When a strain lacking the major isoform of PGM (pgm2Delta) was grown on media containing galactose as sole carbon source, its rate of Ca(2+) uptake was 5-fold higher than an isogenic wild-type strain. This increased rate of Ca(2+) uptake resulted in a 9-fold increase in the steady-state total cellular Ca(2+) level. The fraction of cellular Ca(2+) located in the exchangeable pool in the pgm2Delta strain was found to be as large as the exchangeable fraction observed in wild-type cells, suggesting that the depletion of Golgi Ca(2+) stores is not responsible for the increased rate of Ca(2+) uptake. We also found that growth of the pgm2Delta strain on galactose media is inhibited by 10 microM cyclosporin A, suggesting that activation of the calmodulin/calcineurin signaling pathway is required to activate the Ca(2+) transporters that sequester the increased cytosolic Ca(2+) load caused by this high rate of Ca(2+) uptake. We propose that these Ca(2+)-related alterations are attributable to a reduced metabolic flux between Glc-1-P and Glc-6-P due to a limitation of PGM enzymatic activity in the pgm2Delta strain. Consistent with this hypothesis, we found that this "metabolic bottleneck" resulted in an 8-fold increase in the Glc-1-P level compared with the wild-type strain, while the Glc-6-P and ATP levels were normal. These results suggest that Glc-1-P (or a related metabolite) may participate in the control of Ca(2+) uptake from the environment. PMID- 10681520 TI - Induction of rac-guanine nucleotide exchange activity of Ras-GRF1/CDC25(Mm) following phosphorylation by the nonreceptor tyrosine kinase Src. AB - Ras-GRF1/CDC25(Mm) has been implicated as a Ras-guanine nucleotide exchange factor (GEF) expressed in brain. Ras-GEF activity of Ras-GRF1 is augmented in response to Ca(2+) influx and G protein betagamma subunit (Gbetagamma) stimulation. Ras-GRF1 also acts as a GEF toward Rac, but not Rho and Cdc42, when activated by Gbetagamma-mediated signals. Tyrosine phosphorylation of Ras-GRF1 is critical for the induction of Rac-GEF activity as evidenced by inhibition by tyrosine kinase inhibitors. Herein, we show that the nonreceptor tyrosine kinase Src phosphorylates Ras-GRF1, thereby inducing Rac-GEF activity. Ras-GRF1 transiently expressed with v-Src was tyrosine-phosphorylated and showed significant GEF activity toward Rac, but not Rho and Cdc42, which was comparable with that induced by Gbetagamma. In contrast, Ras-GEF activity remained unchanged. The recombinant c-Src protein phosphorylated affinity-purified glutathione S-transferase-tagged Ras-GRF1 in vitro and thereby elicited Rac-GEF activity. Taken together, tyrosine phosphorylation by Src is sufficient for the induction of Rac-GEF activity of Ras-GRF1, which may imply the involvement of Src downstream of Gbetagamma to regulate Ras-GRF1. PMID- 10681521 TI - Mechanisms by which soluble endothelial cell protein C receptor modulates protein C and activated protein C function. AB - The endothelial cell protein C receptor (EPCR) functions as an important regulator of the protein C anticoagulant pathway by binding protein C and enhancing activation by the thrombin-thrombomodulin complex. EPCR binds to both protein C and activated protein C (APC) with high affinity. A soluble form of EPCR (sEPCR) circulates in plasma and inhibits APC anticoagulant activity. In this study, we investigate the mechanisms by which sEPCR modulates APC function. Soluble EPCR inhibited the inactivation of factor Va by APC only in the presence of phospholipid vesicles. By using flow cytometric analysis in the presence of 3 mM CaCl(2) and 0. 6 mM MgCl(2), sEPCR inhibited the binding of protein C and APC to phospholipid vesicles (K(i) = 40 +/- 7 and 33 +/- 4 nM, respectively). Without MgCl(2), the K(i) values increased approximately 4-fold. Double label flow cytometric analysis using fluorescein-APC and Texas Red-sEPCR indicated that the APC.sEPCR complex does not interact with phospholipid vesicles. By using surface plasmon resonance, we found that sEPCR also inhibited binding of protein C to phospholipid in a dose-dependent fashion (K(i) = 32 nM). To explore the possibility that sEPCR evokes structural changes in APC, fluorescence spectroscopy studies were performed to monitor sEPCR/Fl-APC interactions. sEPCR binds saturably to Fl-APC (K(d) = 27 +/- 13 nM) with a maximum decrease in Fl-APC fluorescence of 10.8 +/- 0.6%. sEPCR also stimulated the amidolytic activity of APC toward synthetic substrates. We conclude that sEPCR binding to APC blocks phospholipid interaction and alters the active site of APC. PMID- 10681522 TI - Dissecting the interaction of SHP-2 with PZR, an immunoglobulin family protein containing immunoreceptor tyrosine-based inhibitory motifs. AB - Tyrosine phosphorylation of membrane proteins plays a crucial role in cell signaling by recruiting Src homology 2 (SH2) domain-containing signaling molecules. Recently, we have isolated a transmembrane protein designated PZR that specifically binds tyrosine phosphatase SHP-2, which has two SH2 domains (Zhao, Z. J., and Zhao, R. (1998) J. Biol. Chem. 273, 29367-29372). PZR belongs to the immunoglobulin superfamily. Its intracellular segment contains four putative sites of tyrosine phosphorylation. By site-specific mutagenesis, we found that the tyrosine 241 and 263 embedded in the consensus immunoreceptor tyrosine-based inhibitory motifs VIYAQL and VVYADI, respectively, accounted for the entire tyrosine phosphorylation of PZR. The interaction between PZR and SHP-2 requires involvement of both tyrosyl residues of the former and both SH2 domains of the latter, since its was disrupted by mutating a single tyrosyl residue or an SH2 domain. Overexpression of catalytically inactive but not active forms of SHP-2 bearing intact SH2 domains in cells caused hyperphosphorylation of PZR. In vitro, tyrosine-phosphorylated PZR was efficiently dephosphorylated by the full-length form of SHP-2 but not by its SH2 domain-truncated form. Together, the data indicate that PZR serves not only as a specific anchor protein of SHP-2 on the plasma membrane but also as a physiological substrate of the enzyme. The coexisting binding and dephosphorylation of PZR by SHP-2 may function to terminate signal transduction initiated by PZR and SHP-2 and to set a threshold for the signal transduction to be initiated. PMID- 10681523 TI - alpha(1A) adrenergic receptor induces eukaryotic initiation factor 4E-binding protein 1 phosphorylation via a Ca(2+)-dependent pathway independent of phosphatidylinositol 3-kinase/Akt. AB - Phosphorylation of the translation repressor eukaryotic initiation factor 4E binding protein 1 (4E-BP1) is thought to be partly responsible for increased protein synthesis induced by growth factors. This study investigated the effect of a G(q)-coupled receptor on protein synthesis and the phosphorylation state and function of 4E-BP1 in Rat-1 fibroblasts expressing the human alpha(1A) adrenergic receptor. Treatment of cells with phenylephrine (PE), a specific alpha(1) adrenergic receptor agonist, increased protein synthesis and induced the phosphorylation of 4E-BP1 and its release from translation initiation factor 4E. Although the PE-induced phosphorylation of 4E-BP1 was blocked by the phosphatidylinositol 3-kinase inhibitor LY294002, neither phosphatidylinositol 3 kinase nor Akt, its downstream effector, is activated in cells treated with PE (Ballou, L. M., Cross, M. E., Huang, S., McReynolds, E. M., Zhang, B. X., and Lin, R. Z., J. Biol. Chem. 275, 4803-4809). The effect of PE on 4E-BP1 phosphorylation was also abolished in cells depleted of intracellular Ca(2+) and in cells pretreated with calmodulin antagonists. By contrast, phosphorylation of 4E-BP1 still occurred in cells in which the Ca(2+)- and diacylglycerol-dependent isoforms of protein kinase C were down-regulated by prolonged exposure to a phorbol ester. We conclude that activation of the alpha(1A) adrenergic receptor in Rat-1 fibroblasts leads to phosphorylation of 4E-BP1 via a pathway that is Ca(2+)- and calmodulin-dependent. Phosphatidylinositol 3-kinase, Akt, and phorbol ester-sensitive protein kinase C isoforms do not appear to be required in this signaling pathway. PMID- 10681524 TI - Human vascular smooth muscle cells possess functional CCR5. AB - CC chemokine receptors are important modulators of inflammation. Although CC chemokine receptors have been found predominantly on leukocytes, recent studies have suggested that vascular smooth muscle cells respond to CC chemokines. We now report that human smooth muscle cells express CCR5, a co-receptor for human immunodeficiency virus. CCR5 mRNA was detectable by RNA blot hybridization in human aortic and coronary artery smooth muscle cells. The cDNA generated by reverse transcription-polymerase chain reaction from aortic smooth muscle cells had 100% identity throughout the entire coding region with the CCR5 cloned from THP-1 cells. By immunohistochemistry, CCR5 and the CCR5 ligand, macrophage inflammatory protein-1beta (MIP-1beta), were detected in smooth muscle cells and macrophages of the atherosclerotic plaque. In smooth muscle cell culture, MIP 1beta induced a significant increase in intracellular calcium concentrations, which was blocked by an antibody to CCR5. In addition, MIP-1beta caused a calcium dependent increase in tissue factor activity. Tissue factor is the initiator of coagulation and is thought to play a key role in arterial thrombosis. These data suggest that human arterial smooth muscle cells express functional CCR5 receptors and MIP-1beta is an agonist for these cells. PMID- 10681525 TI - Identification of the peptide-binding site in the heat shock chaperone/tumor rejection antigen gp96 (Grp94). AB - Heat shock protein (HSP)-peptide complexes from tumor cells elicit specific protective immunity when injected into inbred mice bearing the same specific type of tumor. The HSP-mediated specific immunogenicity also occurs with virus infected cells. The immune response is solely due to endogenous peptides noncovalently bound to HSP. A vesicular stomatitis virus capsid-derived peptide ligand bearing a photoreactive azido group was specifically bound by and cross linked to murine HSP glycoprotein (gp) 96. The peptide-binding site was mapped by specific proteolysis of the cross-links followed by analysis of the cross-linked peptides using a judicious combination of SDS-gel electrophoresis, mass spectrometry, and amino acid sequencing. The minimal peptide-binding site was mapped to amino acid residues 624-630 in a highly conserved region of gp96. A model of the peptide binding pocket of gp96 was constructed based on the known crystallographic structure of major histocompatibility complex class I molecule bound to a similar peptide. The gp96-peptide model predicts that the peptide ligand is held in a groove formed by alpha-helices and lies on a surface consisting of antiparallel beta-sheets. Interestingly, in this model, the peptide binding pocket abuts the dimerization domain of gp96, which may have implications for the extraordinary stability of peptide-gp96 complexes, and for the faithful relay of peptides to major histocompatibility complex class I molecule for antigen presentation. PMID- 10681526 TI - Pairwise electrostatic interactions between alpha-neurotoxins and gamma, delta, and epsilon subunits of the nicotinic acetylcholine receptor. AB - alpha-Neurotoxins bind with high affinity to alpha-gamma and alpha-delta subunit interfaces of the nicotinic acetylcholine receptor. Since this high affinity complex likely involves a van der Waals surface area of approximately 1200 A(2) and 25-35 residues on the receptor surface, analysis of side chains should delineate major interactions and the orientation of bound alpha-neurotoxin. Three distinct regions on the gamma subunit, defined by Trp(55), Leu(119), Asp(174), and Glu(176), contribute to alpha-toxin affinity. Of six charge reversal mutations on the three loops of Naja mossambica mossambica alpha-toxin, Lys(27) - > Glu, Arg(33) --> Glu, and Arg(36) --> Glu in loop II reduce binding energy substantially, while mutations in loops I and III have little effect. Paired residues were analyzed by thermodynamic mutant cycles to delineate electrostatic linkages between the six alpha-toxin charge reversal mutations and three key residues on the gamma subunit. Large coupling energies were found between Arg(33) at the tip of loop II and gammaLeu(119) (-5.7 kcal/mol) and between Lys(27) and gammaGlu(176) (-5.9 kcal/mol). gammaTrp(55) couples strongly to both Arg(33) and Lys(27), whereas gammaAsp(174) couples minimally to charged alpha-toxin residues. Arg(36), despite strong energetic contributions, does not partner with any gamma subunit residues, perhaps indicating its proximity to the alpha subunit. By analyzing cationic, neutral and anionic residues in the mutant cycles, interactions at gamma176 and gamma119 can be distinguished from those at gamma55. PMID- 10681527 TI - Identification and characterization of a PDZ protein that interacts with activin type II receptors. AB - We have identified a mouse PDZ protein that interacts with the activin type IIA receptor (ActRIIA), which we named activin receptor-interacting protein 1 (ARIP1). By using yeast two-hybrid screening, we isolated a cDNA clone of ARIP1 from a mouse brain cDNA library. We detected two forms of ARIP1, ARIP1-long and ARIP1-short, which may be produced by alternative splicing. ARIP1-long had one guanylate kinase domain in the NH(2)-terminal region, followed by two WW domains and five PDZ domains (PDZ1-5). ARIP1-short had a deletion in the NH(2)-terminal region and lacked the guanylate kinase domain. Both forms interacted with ActRIIA through PDZ5. The COOH-terminal residues of ActRIIA (ESSL) agree with a PDZ binding consensus motif, and ARIP1 recognized the consensus sequence. ARIP1 interacts specifically with ActRIIA among the receptors for the transforming growth factor beta family. Interestingly, ARIP1 also interacted with Smad3, which is an activin/transforming growth factor beta intracellular signaling molecule. The mRNA of ARIP1 was more abundant in the brain than in other tissues. Overexpression of ARIP1 controls activin-induced and Smad3-induced transcription in activin-responsive cell lines. These findings suggest that ARIP1 has a significant role in assembling activin signaling molecules at specific subcellular sites and in regulating signal transduction in neuronal cells. PMID- 10681528 TI - Probing subunit interactions in alpha class rat liver glutathione S-transferase with the photoaffinity label glutathionyl S-[4-(succinimidyl)benzophenone]. AB - Glutathionyl S-[4-(succinimidyl)benzophenone] (GS-Succ-BP), an analogue of the product of glutathione and electrophilic substrate, acts as a photoaffinity label of dimeric rat liver glutathione S-transferase (GST), isoenzyme 1-1. A time dependent loss of enzyme activity is observed upon irradiation of the enzyme with long wavelength UV light in the presence of the reagent. The initial rate of inactivation exhibits nonlinear dependence on the concentration of the reagent, characterized by an apparent dissociation constant of the enzyme-reagent complex (K(R)) of 99 +/- 2 microM and k(max) of 0.082 +/- 0.005 min(-1). Protection against this inactivation is provided by the electrophilic substrate (ethacrynic acid), electrophilic substrate analogue (dinitrophenol), and product analogues (S hexylglutathione and p-nitrobenzylglutathione) but not by steroids (Delta(5) androstene-3,17-dione and 17beta-estradiol-3, 17-disulfate). These results suggest that GS-Succ-BP binds and reacts with the enzyme within the xenobiotic substrate binding site, and this reaction site is distinct from the substrate and nonsubstrate steroid binding sites of the enzyme. About 1 mol of reagent is incorporated into 1 mol of enzyme dimer when the enzyme is completely inactivated. Met-208 is the only amino acid target of the reagent, and modification of this residue in one enzyme subunit of the GST 1-1 dimer completely abolishes the enzyme activity of both subunits. In order to evaluate the role of subunit interactions in the Alpha class glutathione S-transferases, inactive GS-Succ-BP-modified GST 1-1 was mixed with unlabeled, active GST 2-2. The enzyme subunits were dissociated in dilute trifluoroacetic acid and then renatured at pH 7.8 and separated by chromatofocusing into GST 1-1, 1-2, and 2-2. The specific activities of the heterodimer toward several substrates indicate that the loss of catalytic activity in the unmodified subunit of the modified GST 1-1 is the indirect result of the interaction between the two enzyme subunits and that this subunit interaction is absent in the heterodimer GST 1-2. PMID- 10681529 TI - Cysteine-rich protein 2, a novel substrate for cGMP kinase I in enteric neurons and intestinal smooth muscle. AB - Nitric oxide/cGMP/cGMP kinase I (cGKI) signaling causes relaxation of intestinal smooth muscle. In the gastrointestinal tract substrates of cGKI have not been identified yet. In the present study a protein interacting with cGKIbeta has been isolated from a rat intestinal cDNA library using the yeast two-hybrid system. The protein was identified as cysteine-rich protein 2 (CRP2), recently cloned from rat brain (Okano, I., Yamamoto, T., Kaji, A., Kimura, T., Mizuno, K., and Nakamura, T. (1993) FEBS Lett. 333, 51-55). Recombinant CRP2 is specifically phosphorylated by cGKs but not by cAMP kinase in vitro. Co-transfection of CRP2 and cGKIbeta into COS cells confirmed the phosphorylation of CRP2 in vivo. Cyclic GMP kinase I phosphorylated CRP2 at Ser-104, because the mutation to Ala completely prevented the in vivo phosphorylation. Immunohistochemical analysis using confocal laser scan microscopy showed a co-localization of CRP2 and cGKI in the inner part of the circular muscle layer, in the muscularis mucosae, and in specific neurons of the myenteric and submucosal plexus. The co-localization together with the specific phosphorylation of CRP2 by cGKI in vitro and in vivo suggests that CRP2 is a novel substrate of cGKI in neurons and smooth muscle of the small intestine. PMID- 10681530 TI - The human AC133 hematopoietic stem cell antigen is also expressed in epithelial cells and targeted to plasma membrane protrusions. AB - The human AC133 antigen and mouse prominin are structurally related plasma membrane proteins. However, their tissue distribution is distinct, with the AC133 antigen being found on hematopoietic stem and progenitor cells and prominin on various epithelial cells. To determine whether the human AC133 antigen and mouse prominin are orthologues or distinct members of a protein family, we examined the human epithelial cell line Caco-2 for the possible expression of the AC133 antigen. By both immunofluorescence and immunoprecipitation, the AC133 antigen was found to be expressed on the surface of Caco-2 cells. Interestingly, immunoreactivity for the AC133 antigen, but not its mRNA level, was down regulated upon differentiation of Caco-2 cells. The AC133 antigen was specifically located at the apical rather than basolateral plasma membrane. An apical localization of the AC133 antigen was also observed in various human embryonic epithelia including the neural tube, gut, and kidney. Electron microscopy revealed that, within the apical plasma membrane of Caco-2 cells, the AC133 antigen was confined to microvilli and absent from the planar, intermicrovillar regions. This specific subcellular localization did not depend on an epithelial phenotype, because the AC133 antigen on hematopoietic stem cells, as well as that ectopically expressed in fibroblasts, was selectively found in plasma membrane protrusions. Hence, the human AC133 antigen shows the features characteristic of mouse prominin in epithelial and transfected non epithelial cells, i.e. a selective association with apical microvilli and plasma membrane protrusions, respectively. Conversely, flow cytometry of murine CD34(+) bone marrow progenitors revealed the cell surface expression of prominin. Taken together, the data strongly suggest that the AC133 antigen is the human orthologue of prominin. PMID- 10681531 TI - X-ray structure of beta-carbonic anhydrase from the red alga, Porphyridium purpureum, reveals a novel catalytic site for CO(2) hydration. AB - The carbonic anhydrases (CAs) fall into three evolutionarily distinct families designated alpha-, beta-, and gamma-CAs based on their primary structure. beta CAs are present in higher plants, algae, and prokaryotes, and are involved in inorganic carbon utilization. Here, we describe the novel x-ray structure of beta CA from the red alga, Porphyridium purpureum, at 2.2-A resolution using intrinsic zinc multiwavelength anomalous diffraction phasing. The CA monomer is composed of two internally repeating structures, being folded as a pair of fundamentally equivalent motifs of an alpha/beta domain and three projecting alpha-helices. The motif is obviously distinct from that of either alpha- or gamma-CAs. This homodimeric CA appears like a tetramer with a pseudo 222 symmetry. The active site zinc is coordinated by a Cys-Asp-His-Cys tetrad that is strictly conserved among the beta-CAs. No water molecule is found in a zinc-liganding radius, indicating that the zinc-hydroxide mechanism in alpha-CAs, and possibly in gamma CAs, is not directly applicable to the case in beta-CAs. Zinc coordination environments of the CAs provide an interesting example of the convergent evolution of distinct catalytic sites required for the same CO(2) hydration reaction. PMID- 10681532 TI - Cys-93-betabeta-succinimidophenyl polyethylene glycol 2000 hemoglobin A. Intramolecular cross-bridging of hemoglobin outside the central cavity. AB - Bis(maleidophenyl)-PEG2000 (Bis-Mal-PEG2000), a new bifunctional protein cross linker targeted to sulfhydryl groups, introduces intra-tetrameric cross-links into oxy-HbA in nearly quantitative yields. Structural as well as crystallographic analyses of the cross-linked species, Bis-Mal-PEG2000 HbA, identified Cys-93(beta) as the site of intramolecular cross-linking. The cross bridging had only a limited influence on the O(2) affinity and cooperativity of HbA in 50 mM BisTris acetate, pH 7.4. However, the Bohr effect was reduced by approximately 60%. Bis-Mal-PEG2000 HbA retained sensitivity to the presence of allosteric effectors 2, 3-diphosphoglycerate, IHP, and chloride, albeit to a lesser degree compared with HbA. Crystallographic analysis revealed the overall structure of deoxy-Bis-Mal-PEG2000 HbA to be similar to deoxy-HbA but for the loss of the salt bridge between Asp-94(beta) and His-146(beta). The large influence of the cross-bridging on the alkaline Bohr effect of HbA is consistent with the loss of this salt bridge. Unlike the "central cavity cross-bridges" described previously, the cross-link introduced by Bis-Mal-PEG2000 into HbA is an "outside the central cavity cross-bridge." In view of its oxy-conformational specificity and limited influence on O(2) affinity, this new cross-linking strategy holds promise for the stabilization of new designer low O(2) affinity Hbs generated by recombinant DNA technology for applications as Hb based therapeutics. PMID- 10681533 TI - Phosphorylation of microtubule-associated protein tau is regulated by protein phosphatase 2A in mammalian brain. Implications for neurofibrillary degeneration in Alzheimer's disease. AB - Hyperphosphorylated tau, which is the major protein of the neurofibrillary tangles in Alzheimer's disease brain, is most probably the result of an imbalance of tau kinase and phosphatase activities in the affected neurons. By using metabolically competent rat brain slices as a model, we found that selective inhibition of protein phosphatase 2A by okadaic acid induced an Alzheimer-like hyperphosphorylation and accumulation of tau. The hyperphosphorylated tau had a reduced ability to bind to microtubules and to promote microtubule assembly in vitro. Immunocytochemical staining revealed hyperphosphorylated tau accumulation in pyramidal neurons in cornu ammonis and in neocortical neurons. The topography of these changes recalls the distribution of neurofibrillary tangles in Alzheimer's disease brain. Selective inhibition of protein phosphatase 2B with cyclosporin A did not have any significant effect on tau phosphorylation, accumulation, or function. These studies suggest that protein phosphatase 2A participates in regulation of tau phosphorylation, processing, and function in vivo. A down-regulation of protein phosphatase 2A activity can lead to Alzheimer like abnormal hyperphosphorylation of tau. PMID- 10681534 TI - Chymase as a proangiogenic factor. A possible involvement of chymase-angiotensin dependent pathway in the hamster sponge angiogenesis model. AB - We investigated the profound involvement of chymase, an alternative angiotensin II-generating enzyme, in angiogenesis using a hamster sponge implant model. In vivo transfection of human pro-chymase cDNA or a direct injection of purified chymase into the sponges implanted resulted in marked increment of hemoglobin contents in the sponge granuloma tissues, demonstrating that chymase has an ability to elicit angiogenesis and is a potent angiogenic factor. Daily injection of basic fibroblast growth factor into the sponges implanted also induced angiogenesis, which was suppressed by the treatment with chymostatin, an inhibitor of chymase, or TCV-116, an antagonist of angiotensin II (Ang II) type 1 receptor. Expression of chymase mRNA and production of Ang II in the granuloma tissues were enhanced by the stimulation with basic fibroblast growth factor. Chymase activity in the sponge granulomas increased in parallel with the rise in hemoglobin contents, and mast cells observed in the granuloma tissues were positively stained with anti-chymase antibody. Exogenous administration not only of Ang II but of angiotensin I (Ang I) directly into the sponges could enhance angiogenesis. Chymostatin inhibited the angiogenesis induced by Ang I but not Ang II, suggesting the presence of a chymase-like Ang II-generating activity in the sponge granulomas. Our results may suggest a potential ability of chymase to promote angiogenesis through the local chymase-dependent and angiotensin converting enzyme-dependent Ang II generating system in pathophysiological angiogenesis. PMID- 10681535 TI - Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of a human thrombopoietin-dependent hematopoietic cell line. AB - At the late phase of megakaryocytopoiesis, megakaryocytes undergo endomitosis, which is characterized by DNA replication without cell division. Although a number of cell cycle regulatory molecules have been identified, the precise roles of these molecules in megakaryocytic endomitosis are largely unknown. In a human interleukin-3-dependent cell line transfected with the thrombopoietin (TPO) receptor c-mpl (F-36P-mpl), either treatment with TPO or the overexpression of activated ras (Ha-Ras(G12V)) induced megakaryocytic maturation with polyploid formation. We found that TPO stimulation or Ha-Ras(G12V) expression led to up regulation of cyclin D1, cyclin D2, and cyclin D3 expression. In addition, expression levels of cyclin A and cyclin B were reduced during the total course of both TPO- and Ha-Ras(G12V)-induced megakaryocytic differentiation, thereby leading to decreased cdc2 kinase activity. Neither the induced expression of cyclin D1, cyclin D2, or cyclin D3 nor the expression of a dominant negative form of cdc2 alone could induce megakaryocytic differentiation of F-36P-mpl cells. In contrast, overexpression of dominant negative cdc2 together with cyclin D1, cyclin D2, or cyclin D3 facilitated megakaryocytic differentiation in the absence of TPO. These results suggest that both D-type cyclin expression and decreased cdc2 kinase activity may participate in megakaryocytic differentiation. PMID- 10681536 TI - Functional expression of the human hZIP2 zinc transporter. AB - Zinc is an essential nutrient for humans, yet we know little about how this metal ion is taken up by mammalian cells. In this report, we describe the characterization of hZip2, a human zinc transporter identified by its similarity to zinc transporters recently characterized in fungi and plants. hZip2 is a member of the ZIP family of eukaryotic metal ion transporters that includes two other human genes, hZIP1 and hZIP3, and genes in mice and rats. To test whether hZip2 is a zinc transporter, we examined (65)Zn uptake activity in transfected K562 erythroleukemia cells expressing hZip2 from the CMV promoter. hZip2 expressing cells accumulated more zinc than control cells because of an increased initial zinc uptake rate. This activity was time-, temperature-, and concentration-dependent and saturable with an apparent K(m) of 3 microM. hZip2 zinc uptake activity was inhibited by several other transition metals, suggesting that this protein may transport other substrates as well. hZip2 activity was not energy-dependent, nor did it require K(+) or Na(+) gradients. Zinc uptake by hZip2 was stimulated by HCO(3)(-) treatment, suggesting a Zn(2+)-HCO(3)(-) cotransport mechanism. Finally, hZip2 was exclusively localized in the plasma membrane. These results indicate that hZip2 is a zinc transporter, and its identification provides one of the first molecular tools to study zinc uptake in mammalian cells. PMID- 10681537 TI - Recognition of misfolding proteins by PA700, the regulatory subcomplex of the 26 S proteasome. AB - The 26 S proteasome is a large protease complex that catalyzes the degradation of both native and misfolded proteins. These proteins are known to interact with PA700, the regulatory subcomplex of the 26 S proteasome, via a covalently attached polyubiquitin chain. Here we provide evidence for an additional ubiquitin-independent mode of substrate recognition by PA700. PA700 prevents the aggregation of three incompletely folded, nonubiquitinated substrates: the DeltaF 508 mutant form of cystic fibrosis transmembrane regulator, nucleotide binding domain 1, insulin B chain, and citrate synthase. This function does not require ATP hydrolysis. The stoichiometry required for this function, the effect of PA700 on the lag phase of aggregation, and the temporal specificity of PA700 in this process all indicate that PA700 interacts with a subpopulation of non-native conformations that is either particularly aggregation-prone or nucleates misassociation reactions. The inhibition of off-pathway self-association reactions is also reflected in the ability of PA700 to promote refolding of citrate synthase. These results provide evidence that, in addition to binding polyubiquitin chains, PA700 contains a site(s) that recognizes and interacts with misfolded or partially denatured polypeptides. This feature supplies an additional level of substrate specificity to the 26 S proteasome and a means by which substrates are maintained in a soluble state until refolding or degradation is complete. PMID- 10681538 TI - The yeast STM1 gene encodes a purine motif triple helical DNA-binding protein. AB - The formation of triple helical DNA has been evoked in several cellular processes including transcription, replication, and recombination. Using conventional and affinity chromatography, we purified from Saccharomyces cerevisiae whole-cell extract a 35-kDa protein that avidly and specifically bound a purine motif triplex (with a K(d) of 61 pM) but not a pyrimidine motif triplex or duplex DNA. Peptide microsequencing identified this protein as the product of the STM1 gene. Confirmation that Stm1p is a purine motif triplex-binding protein was obtained by electrophoretic mobility shift assays using either bacterially expressed, recombinant Stm1p or whole-cell extracts from stm1Delta yeast. Stm1p has previously been identified as G4p2, a G-quartet nucleic acid-binding protein. This suggests that some proteins actually recognize features shared by G4 DNA and purine motif triplexes, e.g. Hoogsteen hydrogen-bonded guanines. Genetically, the STM1 gene has been identified as a multicopy suppressor of mutations in several genes involved in mitosis (e.g. TOM1, MPT5, and POP2). A possible role for multiplex DNA and its binding proteins in mitosis is discussed. PMID- 10681539 TI - Molecular characterization of the non-biotin-containing subunit of 3 methylcrotonyl-CoA carboxylase. AB - The biotin enzyme, 3-methylcrotonyl-CoA carboxylase (MCCase) (3-methylcrotonyl CoA:carbon-dioxide ligase (ADP-forming), EC 6.4.1. 4), catalyzes a pivotal reaction required for both leucine catabolism and isoprenoid metabolism. MCCase is a heteromeric enzyme composed of biotin-containing (MCC-A) and non-biotin containing (MCC-B) subunits. Although the sequence of the MCC-A subunit was previously determined, the primary structure of the MCC-B subunit is unknown. Based upon sequences of biotin enzymes that use substrates structurally related to 3-methylcrotonyl-CoA, we isolated the MCC-B cDNA and gene of Arabidopsis. Antibodies directed against the bacterially produced recombinant protein encoded by the MCC-B cDNA react solely with the MCC-B subunit of the purified MCCase and inhibit MCCase activity. The primary structure of the MCC-B subunit shows the highest similarity to carboxyltransferase domains of biotin enzymes that use methyl-branched thiol esters as substrate or products. The single copy MCC-B gene of Arabidopsis is interrupted by nine introns. MCC-A and MCC-B mRNAs accumulate in all cell types and organs, with the highest accumulation occurring in rapidly growing and metabolically active tissues. In addition, these two mRNAs accumulate coordinately in an approximately equal molar ratio, and they each account for between 0.01 and 0.1 mol % of cellular mRNA. The sequence of the Arabidopsis MCC B gene has enabled the identification of animal paralogous MCC-B cDNAs and genes, which may have an impact on the molecular understanding of the lethal inherited metabolic disorder methylcrotonylglyciuria. PMID- 10681540 TI - Protein kinase C activation stimulates the phosphorylation and internalization of the sst2A somatostatin receptor. AB - The sst2A receptor is expressed in the endocrine, gastrointestinal, and neuronal systems as well as in many hormone-sensitive tumors. This receptor is rapidly internalized and phosphorylated in growth hormone-R2 pituitary cells following somatostatin binding (Hipkin, R. W., Friedman, J., Clark, R. B., Eppler, C. M., and Schonbrunn, A. (1997) J. Biol. Chem. 272, 13869-13876). The protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), also stimulates sst2A phosphorylation. Here we examine the mechanisms and consequences of PMA and agonist-induced sst2A phosphorylation. Like somatostatin, both PMA and bombesin increased sst2A receptor phosphorylation within 2 min. The PKC inhibitor GF109203X blocked PMA- and bombesin- stimulated sst2A phosphorylation, whereas stimulation by the somatostatin analog SMS 201-995 was unaffected. Agonist and PMA each stimulated phosphorylation in two receptor domains, the third intracellular loop and the C-terminal tail. Functionally, PMA dramatically increased the internalization of the sst2A receptor-ligand complex. This PMA stimulation was blocked by GF109203X, whereas basal internalization was unaffected. However, neither basal nor PMA-stimulated internalization was altered by pertussis toxin, whereas both were blocked by hypertonic sucrose. Therefore PKC activation and agonist binding stimulate sst2A phosphorylation by distinct mechanisms, and PKC potentiates internalization of the sst2A receptor via clathrin-coated pits. Thus, hormonal stimulation of PKC-coupled receptors may provide a mechanism for regulating the inhibitory actions of somatostatin in target tissue. PMID- 10681541 TI - The Chk1 protein kinase and the Cdc25C regulatory pathways are targets of the anticancer agent UCN-01. AB - A checkpoint operating in the G(2) phase of the cell cycle prevents entry into mitosis in the presence of DNA damage. UCN-01, a protein kinase inhibitor currently undergoing clinical trials for cancer treatment, abrogates G(2) checkpoint function and sensitizes p53-defective cancer cells to DNA-damaging agents. In most species, the G(2) checkpoint prevents the Cdc25 phosphatase from removing inhibitory phosphate groups from the mitosis-promoting kinase Cdc2. This is accomplished by maintaining Cdc25 in a phosphorylated form that binds 14-3-3 proteins. The checkpoint kinases, Chk1 and Cds1, are proposed to regulate the interactions between human Cdc25C and 14-3-3 proteins by phosphorylating Cdc25C on serine 216. 14-3-3 proteins, in turn, function to keep Cdc25C out of the nucleus. Here we report that UCN-01 caused loss of both serine 216 phosphorylation and 14-3-3 binding to Cdc25C in DNA-damaged cells. In addition, UCN-01 potently inhibited the ability of Chk1 to phosphorylate Cdc25C in vitro. In contrast, Cds1 was refractory to inhibition by UCN-01 in vitro, and Cds1 was still phosphorylated in irradiated cells treated with UCN-01. Thus, neither Cds1 nor kinases upstream of Cds1, such as ataxia telangiectasia-mutated, are targets of UCN-01 action in vivo. Taken together our results identify the Chk1 kinase and the Cdc25C pathway as potential targets of G(2) checkpoint abrogation by UCN-01. PMID- 10681542 TI - Identification by mutagenesis of arginines in the substrate binding site of the porcine NADP-dependent isocitrate dehydrogenase. AB - Pig heart mitochondrial NADP-dependent isocitrate dehydrogenase is the most extensively studied among the mammalian isocitrate dehydrogenases. The crystal structure of Escherichia coli isocitrate dehydrogenase and sequence alignment of porcine with E. coli isocitrate dehydrogenase suggests that the porcine Arg(101), Arg(110), Arg(120), and Arg(133) are candidates for roles in substrate binding. The four arginines were separately mutated to glutamine using a polymerase chain reaction method. Wild type and mutant enzymes were each expressed in E. coli, isolated as maltose binding fusion proteins, then cleaved with thrombin, and purified to yield homogeneous porcine isocitrate dehydrogenase. The R120Q mutant has a specific activity, as well as K(m) values for isocitrate, Mn(2+), and NADP(+) similar to wild type enzyme, indicating that Arg(120) is not needed for function. The specific activities of R101Q, R110Q, and R133Q are 1.73, 1.30, and 19.7 micromols/min/mg, respectively, as compared with 39.6 units/mg for wild type enzyme. The R110Q and R133Q enzymes exhibit K(m) values for isocitrate that are increased more than 400- and 165-fold, respectively, as compared with wild type. The K(m) values for Mn(2+), but not for NADP(+), are also elevated indicating that binding of the metal-isocitrate complex is impaired in these mutants. It is proposed that the positive charges of Arg(110) and Arg(133) normally strengthen the binding of the negatively charged isocitrate by electrostatic attraction. The R101Q mutant shows smaller, but significant increases in the K(m) values for isocitrate and Mn(2+); however, the marked decrease in k(cat) suggests a role for Arg(101) in catalysis. The V(max) of wild type enzyme depends on the ionized form of an enzymic group of pK 5.5, and this pK(aes) is similar for the R101Q and R120Q enzymes. In contrast, the pK(aes) for R110Q and R133Q enzymes increases to 6.4 and 7.4, respectively, indicating that the positive charges of Arg(110) and Arg(133) normally lower the pK of the nearby catalytic base to facilitate its ionization. These results may be understood in terms of the structure of the porcine NADP-specific isocitrate dehydrogenase generated by the Insight II Modeler Program, based on the x-ray coordinates of the E. coli enzyme. PMID- 10681543 TI - Opposing role of mitogen-activated protein kinase subtypes, erk-1/2 and p38, in the regulation of chondrogenesis of mesenchymes. AB - The present studies were performed to determine subtype-specific roles of mitogen activated protein kinase in chondrogenesis. Erk-1/2 activities, downstream of protein kinase C, decreased as chondrogenesis proceeded, whereas p38 activities, independent of protein kinase C, continuously increased during chondrogenesis. Inhibition of Erk-1/2 with PD98059 enhanced chondrogenesis up to 1. 7-fold, whereas inhibition of p38 with SB203580 reduced it to about 30% of the control level. Inhibition of Erk-1/2 or p38 did not affect precartilage condensation. However, cartilage nodule formation was significantly blocked by the inhibition of p38, whereas Erk-1/2 inhibition did not affect it. Modulation of chondrogenesis by the inhibition of Erk-1/2 and p38 was accompanied by altered expression of adhesion molecules in an opposite way. Expression of N-cadherin was reduced as chondrogenesis proceeded. Inhibition of p38 caused sustained expression of N-cadherin, whereas Erk-1/2 inhibition accelerated the reduction of N-cadherin expression. Expression of integrin alpha5beta1 and fibronectin were found to transiently increase during chondrogenesis. Inhibition of p38 caused continuous increase of expression of these molecules, whereas Erk-1/2 inhibition accelerated the decrease of expression of these molecules at a later period of chondrogenesis. Because temporal expression of these adhesion molecules regulates chondrogenesis, the above results indicate that Erk-1/2 and p38 conversely regulate chondrogenesis at post-precartilage condensation stages by modulating expression of adhesion molecules. PMID- 10681544 TI - The role of tryptophan residues in the 5-Hydroxytryptamine(3) receptor ligand binding domain. AB - Aromatic amino acids are important components of the ligand binding site in the Cys loop family of ligand-gated ion channels. To examine the role of tryptophan residues in the ligand binding domain of the 5-hydroxytryptamine(3) (5-HT(3)) receptor, we used site-directed mutagenesis to change each of the eight N terminal tryptophan residues in the 5-HT(3A) receptor subunit to tyrosine or serine. The mutants were expressed as homomeric 5-HT(3A) receptors in HEK293 cells and analyzed with radioligand binding, electrophysiology, and immunocytochemistry. Mutation of Trp(90), Trp(183), and Trp(195) to tyrosine resulted in functional receptors, although with increased EC(50) values (2-92 fold) to 5-HT(3) receptor agonists. Changing these residues to serine either ablated function (Trp(90) and Trp(183)) or resulted in a further increase in EC(50) (Trp(195)). Mutation of residue Trp(60) had no effect on ligand binding or receptor function, whereas mutation of Trp(95), Trp(102), Trp(121), and Trp(214) ablated ligand binding and receptor function, and all but one of the receptors containing these mutations were not expressed at the plasma membrane. We propose that Trp(90), Trp(183), and Trp(195) are intimately involved in ligand binding, whereas Trp(95), Trp(102), Trp(121), and Trp(214) have a critical role in receptor structure or assembly. PMID- 10681545 TI - beta-Amyloid(1-42) binds to alpha7 nicotinic acetylcholine receptor with high affinity. Implications for Alzheimer's disease pathology. AB - Alzheimer's disease pathology is characterized by the presence of neuritic plaques and the loss of cholinergic neurons in the brain. The underlying mechanisms leading to these events are unclear, but the 42-amino acid beta amyloid peptide (Abeta(1-42)) is involved. Immunohistochemical studies on human sporadic Alzheimer's disease brains demonstrate that Abeta(1-42) and a neuronal pentameric cation channel, the alpha7 nicotinic acetylcholine receptor (alpha7nAChR), are both present in neuritic plaques and co-localize in individual cortical neurons. Using human brain tissues and cells that overexpress either alpha7nAChR or amyloid precursor protein as the starting material, Abeta(1-42) and alpha7nAChR can be co-immunoprecipitated by the respective specific antibodies, suggesting that they are tightly associated. The formation of the alpha7nAChR.Abeta(1-42) complex can be efficiently suppressed by Abeta(12-28), implying that this Abeta sequence region contains the binding epitope. Receptor binding experiments show that Abeta(1-42) and alpha7nAChR bind with high affinity, and this interaction can be inhibited by alpha7nAChR ligands. Human neuroblastoma cells overexpressing alpha7nAChR are readily killed by Abeta(1-42), whereas alpha7nAChR agonists such as nicotine and epibatidine offered protection. Because Abeta(1-42) inhibits alpha7nAChR-dependent calcium activation and acetylcholine release, two processes critically involved in memory and cognitive functions, and the distribution of alpha7nAChR correlates with neuritic plaques in Alzheimer's disease brains, we propose that interaction of the alpha7nAChR and Abeta(1-42) is a pivotal mechanism involved in the pathophysiology of Alzheimer's disease. PMID- 10681546 TI - Phenylethylthiazolylthiourea (PETT) non-nucleoside inhibitors of HIV-1 and HIV-2 reverse transcriptases. Structural and biochemical analyses. AB - Most non-nucleoside reverse transcriptase (RT) inhibitors are specific for HIV-1 RT and demonstrate minimal inhibition of HIV-2 RT. However, we report that members of the phenylethylthiazolylthiourea (PETT) series of non-nucleoside reverse transcriptase inhibitors showing high potency against HIV-1 RT have varying abilities to inhibit HIV-2 RT. Thus, PETT-1 inhibits HIV-1 RT with an IC(50) of 6 nM but shows only weak inhibition of HIV-2 RT, whereas PETT-2 retains similar potency against HIV-1 RT (IC(50) of 5 nM) and also inhibits HIV-2 RT (IC(50) of 2.2 microM). X-ray crystallographic structure determinations of PETT-1 and PETT-2 in complexes with HIV-1 RT reveal the compounds bind in an overall similar conformation albeit with some differences in their interactions with the protein. To investigate whether PETT-2 could be acting at a different site on HIV 2 RT (e.g. the dNTP or template primer binding site), we compared modes of inhibition for PETT-2 against HIV-1 and HIV-2 RT. PETT-2 was a noncompetitive inhibitor with respect to the dGTP substrate for both HIV-1 and HIV-2 RTs. PETT-2 was also a noncompetitive inhibitor with respect to a poly(rC).(dG) template primer for HIV-2 RT. These results are consistent with PETT-2 binding in corresponding pockets in both HIV-1 and HIV-2 RT with amino acid sequence differences in HIV-2 RT affecting the binding of PETT-2 compared with PETT-1. PMID- 10681547 TI - Identification and characterization of a novel hepatic canalicular ATP diphosphohydrolase. AB - We have identified and characterized a novel ATP diphosphohydrolase (ATPDase) with features of E-type ATPases from porcine liver. Immunoblotting with a specific monoclonal antibody to this ectoenzyme revealed high expression in liver with lesser amounts in kidney and duodenum. This ATPDase was localized by immunohistochemistry to the bile canalicular domain of hepatocytes and to the luminal side of the renal ductular epithelium. In contrast, ATPDase/cd39 was detected in vascular endothelium and smooth muscle in these organs. We purified the putative ATPDase from liver by immunoaffinity techniques and obtained a heavily glycosylated protein with a molecular mass estimated at 75 kDa. This enzyme hydrolyzed all tri- and diphosphonucleosides but not AMP or diadenosine polyphosphates. There was an absolute requirement for divalent cations (Ca(2+) > Mg(2+)). Biochemical activity was unaffected by sodium azide or other inhibitors of ATPases. Kinetic parameters derived from purified preparations of hepatic ATPDase indicated V(max) of 8.5 units/mg of protein with apparent K(m) of 100 microM for both ATP or ADP as substrates. NH(2)-terminal amino acid sequencing revealed near 50% identity with rat liver lysosomal (Ca(2+)-Mg(2+))-ATPase. The different biochemical properties and localization of the hepatic ATPDase suggest pathophysiological functions that are distinct from the vascular ATPDase/cd39. PMID- 10681548 TI - Identification of a gp130 cytokine receptor critical site involved in oncostatin M response. AB - Gp130 cytokine receptor is involved in the formation of multimeric functional receptors for interleukin-6 (IL-6), IL-11, leukemia inhibitory factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor, and cardiotrophin-1. Cloning of the epitope recognized by an OSM-neutralizing anti-gp130 monoclonal antibody identified a portion of gp130 receptor localized in the EF loop of the cytokine binding domain. Site-directed mutagenesis of the corresponding region was carried out by alanine substitution of residues 186-198. To generate type 1 or type 2 OSM receptors, gp130 mutants were expressed together with either LIF receptor beta or OSM receptor beta. When positions Val-189/Tyr-190 and Phe-191/Val-192 were alanine-substituted, Scatchard analyses indicated a complete abrogation of OSM binding to both type receptors. Interestingly, binding of LIF to type 1 receptor was not affected, corroborating the notion that in this case gp130 mostly behaves as a converter protein rather than a binding receptor. The present study demonstrates that positions 189-192 of gp130 cytokine binding domain are essential for OSM binding to both gp130/LIF receptor beta and gp130/OSM receptor beta heterocomplexes. PMID- 10681549 TI - TspO of rhodobacter sphaeroides. A structural and functional model for the mammalian peripheral benzodiazepine receptor. AB - The function and specific structural aspects of the tryptophan-rich sensory protein (TspO) of Rhodobacter sphaeroides 2.4.1 were studied using site-directed mutagenesis involving some 17 different amino acids. The choice of these amino acids changes was dictated from an analysis of the TspO family of proteins derived from the data bases. These studies demonstrated the importance of several highly conserved tryptophan residues in the sensory transduction pathway involving TspO through the proposed binding of an intermediate(s) in the tetrapyrrole biosynthesis pathway. These studies also revealed that the substitution of one or several of the amino acid residues dramatically affected, either directly or indirectly, the levels of TspO in the membranes of R. sphaeroides. Mounting evidence is presented suggesting that TspO normally forms a dimer within the bacterial outer membrane, and the dimer form of TspO may be the active form for TspO function. Because our earlier studies provided us with a functional framework within which to view these amino acid substitutions, we are able to suggest a preliminary model for TspO structure-function. Not only do these studies tell us more about TspO, but they also shed light on the TspO homologue, the drug-binding component of the mitochondrial peripheral benzodiazepine receptor. Mounting evidence draws numerous parallelism between these proteins and supports the significance of using TspO as a model for the structure and function of the mitochondrial protein. PMID- 10681550 TI - Highly hydrophilic proteins in prokaryotes and eukaryotes are common during conditions of water deficit. AB - The late embryogenesis abundant (LEA) proteins are plant proteins that are synthesized at the onset of desiccation in maturing seeds and in vegetative organs exposed to water deficit. Here, we show that most LEA proteins are comprised in a more widespread group, which we call "hydrophilins." The defining characteristics of hydrophilins are high glycine content (>6%) and a high hydrophilicity index (>1.0). By data base searching, we show that this criterion selectively differentiates most known LEA proteins as well as additional proteins from different taxons. We found that within the genomes of Escherichia coli and Saccharomyces cerevisiae, only 5 and 12 proteins, respectively, meet our criterion. Despite their deceivingly loose definition, hydrophilins usually represent <0.2% of the proteins of a genome. Additionally, we demonstrate that the criterion that defines hydrophilins seems to be an excellent predictor of responsiveness to hyperosmosis since most of the genes encoding these proteins in E. coli and S. cerevisiae are induced by osmotic stress. Evidence for the participation of one of the E. coli hydrophilins in the adaptive response to hyperosmotic conditions is presented. Apparently, hydrophilins represent analogous adaptations to a common problem in such diverse taxons as prokaryotes and eukaryotes. PMID- 10681551 TI - The sequence of a gastropod hemocyanin (HtH1 from Haliotis tuberculata). AB - The eight functional units (FUs), a-h, of the hemocyanin isoform HtH1 from Haliotis tuberculata (Prosobranchia, Archaeogastropoda) have been sequenced via cDNA, which provides the first complete primary structure of a gastropod hemocyanin subunit. With 3404 amino acids (392 kDa) it is the largest polypeptide sequence ever obtained for a respiratory protein. The cDNA comprises 10,758 base pairs and includes the coding regions for a short signal peptide, the eight different functional units, a 3'-untranslated region of 478 base pairs, and a poly(A) tail. The predicted protein contains 13 potential sites for N-linked carbohydrates (one for HtH1-a, none for HtH1-c, and two each for the other six functional units). Multiple sequence alignments show that the fragment HtH1 abcdefg is structurally equivalent to the seven-FU subunit from Octopus hemocyanin, which is fundamental to our understanding of the quaternary structures of both hemocyanins. Using the fossil record of the gastropod cephalopod split to calibrate a molecular clock, the origin of the molluscan hemocyanin from a single-FU protein was calculated as 753 +/- 68 million years ago. This fits recent paleontological evidence for the existence of rather large mollusc-like species in the late Precambrian. PMID- 10681552 TI - A network of conserved intramolecular contacts defines the off-state of the transmembrane switch mechanism in a seven-transmembrane receptor. AB - Activation of the rhodopsin-like 7-transmembrane (7-TM) receptors requires switching interhelical constraints that stabilize the inactive state to a new set of contacts in the activated state, which binds the cognate G-protein. The free energy to drive this is provided by agonist binding, which has higher affinity to the active than to the inactive conformation. We have sought specific interhelical constraint contacts, using the M(1) muscarinic acetylcholine receptor as a model. Histidine substitutions of particular groups of amino acids, in transmembrane domains 3, 6, and 7, created high-affinity Zn(2+) binding sites, demonstrating the close proximity of their side chains in the inactive state. Alanine point substitutions have shown the effect of weakening the individual intramolecular contacts. In each case, the acetylcholine affinity was increased, implying promotion of the activated state. These amino acids are highly conserved throughout the 7-TM receptor superfamily. We propose that they form an important part of a network of conserved interhelical contacts that defines the off-state of a general transmembrane switch mechanism. PMID- 10681553 TI - Four conserved cytoplasmic sequence motifs are important for transport function of the Leishmania inositol/H(+) symporter. AB - The protozoan Leishmania donovani has a myo-inositol/proton symporter (MIT) that is a member of a large sugar transporter superfamily. Active transport by MIT is driven by the proton electrochemical gradient across the parasite membrane, and MIT is a prototype for understanding the function of an active transporter in lower eukaryotes. MIT contains two duplicated 6- or 7-amino acid motifs within cytoplasmic loops, which are highly conserved among 50 members of the sugar transporter superfamily and are designated A(1), A(2) ((V)(D/E)(R/K)PhiGR(R/K)), and B(1) (PESPRPhiL), B(2) (VPETKG). In particular, the three acidic residues within these motifs, Glu(187)(B(1)), Asp(300)(A(2)), and Glu(429)(B(2)) in MIT, are highly conserved with 96, 78, and 96% amino acid identity within the analyzed members of this transporter superfamily ranging from bacteria, archaea, and fungi to plants and the animal kingdom. We have used site-directed mutagenesis in combination with functional expression of transporter mutants in Xenopus oocytes and overexpression in Leishmania transfectants to investigate the significance of these three acidic residues in the B(1), A(2), and B(2) motifs. Alteration to the uncharged amides greatly reduced MIT transport function to 23% (E187Q), 1.4% (D300N), and 3% (E429Q) of wild-type activity, respectively, by affecting V(max) but not substrate affinity. Conservative mutations that retained the charge revealed a less pronounced effect on inositol transport with 39% (E187D), 16% (D300E) and 20% (E429D) remaining transport activity. Immunofluorescence microscopy of oocyte cryosections confirmed that MIT mutants were expressed on the oocyte surface in similar quantity to MIT wild type. The proton uncouplers carbonylcyanide-4-(trifluoromethoxy) phenylhydrazone and dinitrophenol inhibited inositol transport by 50-70% in the wild type as well as in E187Q, D300N, and E429Q, despite their reduced transport activities, suggesting that transport in these mutants is still proton-coupled. Furthermore, temperature-dependent uptake studies showed an increased Arrhenius activation energy for the B(1)-E187Q and the B(2)-E429Q mutants, which supports the idea of an impaired transporter cycle in these mutants. We conclude that the conserved acidic residues Glu(187), Asp(300), and Glu(429) are critical for transport function of MIT. PMID- 10681554 TI - Lipoprotein lipase (LPL) strongly links native and oxidized low density lipoprotein particles to decorin-coated collagen. Roles for both dimeric and monomeric forms of LPL. AB - Low density lipoprotein (LDL) and oxidized LDL are associated with collagen in the arterial intima, where the collagen is coated by the small proteoglycan decorin. When incubated in physiological ionic conditions, decorin-coated collagen bound only small amounts of native and oxidized LDL, the interaction being weak. When decorin-coated collagen was first allowed to bind lipoprotein lipase (LPL), binding of native and oxidized LDL increased dramatically (23- and 7-fold, respectively). This increase depended on strong interactions between LPL that was bound to the glycosaminoglycan chains of the collagen-bound decorin and native and oxidized LDL (kDa 12 and 5.9 nM, respectively). To distinguish between binding to monomeric (inactive) and dimeric (catalytically active) forms of LPL, affinity chromatography on heparin columns was conducted, which showed that native LDL bound to the monomeric LPL, whereas oxidized LDL, irrespective of the type of modification (Cu(2+), 2, 2'-azobis(2-amidinopropane)hydrochloride, hypochlorite, or soybean 15-lipoxygenase), bound preferably to dimeric LPL. However, catalytic activity of LPL was not required for binding to oxidized LDL. Finally, immunohistochemistry of atherosclerotic lesions of human coronary arteries revealed specific areas in which LDL, LPL, decorin, and collagen type I were present. The results suggest that LPL can retain LDL in atherosclerotic lesions along decorin-coated collagen fibers. PMID- 10681555 TI - A role for a helical connector between two receptor binding sites of a long-chain peptide hormone. AB - The conformational freedom of single-chain peptide hormones, such as the 41-amino acid hormone corticotropin releasing factor (CRF), is a major obstacle to the determination of their biologically relevant conformation, and thus hampers insights into the mechanism of ligand-receptor interaction. Since N- and C terminal truncations of CRF lead to loss of biological activity, it has been thought that almost the entire peptide is essential for receptor activation. Here we show the existence of two segregated receptor binding sites at the N and C termini of CRF, connection of which is essential for receptor binding and activation. Connection of the two binding sites by highly flexible epsilon aminocaproic acid residues resulted in CRF analogues that remained full, although weak agonists (EC(50): 100-300 nM) independent of linker length. Connection of the two sites by an appropriate helical peptide led to a very potent analogue, which adopted, in contrast to CRF itself, a stable, monomer conformation in aqueous solution. Analogues in which the two sites were connected by helical linkers of different lengths were potent agonists; their significantly different biopotencies (EC(50): 0.6-50 nM), however, suggest the relative orientation between the two binding sites rather than the maintenance of a distinct distance between them to be essential for a high potency. PMID- 10681556 TI - Characterization and subcellular localization of murine and human magnesium dependent neutral sphingomyelinase. AB - Sphingomyelinases (SMases) catalyze the hydrolysis of sphingomyelin, an essential lipid constituent of the plasma membrane, lysosomal membranes, endoplasmic reticulum, and the Golgi membrane stacks of mammalian cells. In this study, we report the biochemical and functional characterization and subcellular localization of magnesium-dependent nSMase1 from overexpressing human embryonic kidney (HEK293) cells. Site-directed mutagenesis of conserved residues probably involved in the enzymatic sphingomyelin cleavage as well as the removal of one or both putative transmembrane domains lead to the complete loss of enzymatic activity of human nSMase1 expressed in HEK293 cells. Polyclonal antibodies raised against recombinant mammalian nSMase1 immunoprecipitated and inactivated the enzyme in membrane extracts of overexpressing HEK293 cells and different murine tissues. Cell fractionation combined with immunoprecipitation studies localized the nSMase1 protein predominantly in the microsomal fraction. The enzyme colocalized with marker proteins of the endoplasmic reticulum and the Golgi apparatus in immunocytochemistry. Anti-nSMase1 antibodies did not affect the nSMase activity in the plasma membrane fraction and membrane extracts from murine brain. Our study leads to the conclusion that nSMase1 is one of at least two mammalian neutral sphingomyelinases with different subcellular localization, tissue specificity, and enzymatic properties. PMID- 10681557 TI - Functional reconstitution of the Na(+)-driven polar flagellar motor component of Vibrio alginolyticus. AB - The bacterial flagellar motor is a molecular machine that couples the influx of specific ions to the generation of the force necessary to drive rotation of the flagellar filament. Four integral membrane proteins, PomA, PomB, MotX, and MotY, have been suggested to be directly involved in torque generation of the Na(+) driven polar flagellar motor of Vibrio alginolyticus. In the present study, we report the isolation of the functional component of the torque-generating unit. The purified protein complex appears to consist of PomA and PomB and contains neither MotX nor MotY. The PomA/B protein, reconstituted into proteoliposomes, catalyzed (22)Na(+) influx in response to a potassium diffusion potential. Sodium uptake was abolished by the presence of Li(+) ions and phenamil, a sodium channel blocker. This is the first demonstration of a purification and functional reconstitution of the bacterial flagellar motor component involved in torque generation. In addition, this study demonstrates that the Na(+)-driven motor component, PomA and PomB, forms the Na(+)-conducting channel. PMID- 10681558 TI - Distinct physiological functions of thiol peroxidase isoenzymes in Saccharomyces cerevisiae. AB - A new type of peroxidase ("thiol peroxidase"; TPx) having cysteine as the primary site of catalysis has been discovered from prokaryotes to eukaryotes. In addition to two yeast TPx isoforms (TSA I and TSA II/AHPC1) previously described, three additional TPx homologues were identified by analysis of the open reading frame data base for Saccharomyces cerevisiae. Three novel isoforms showed a distinct thiol peroxidase activity supported by thioredoxin, and appeared to be distinctively localized in cytoplasm, mitochondria, and nucleus. Each isoform was named after its subcellular localization such as cytoplasmic TPx I (cTPx I or TSA I), cTPx II, cTPx III (TSA II/AHPC1), mitochondrial TPx (mTPx), and nuclear TPx (nTPx). Their transcriptional activities suggest that cTPx I and cTPx III are the most predominant isoforms among the five type isoforms. Transcriptional activities of TPx isoenzymes during yeast life span were quite different from each other. Unlike other TPx null mutants, cTPx I null mutant was hypersensitive to various oxidants except for 4-nitroquinoline N-oxide. The null mutant was more resistant toward 4-nitroquinoline N-oxide and acidic culture than its wild type. The severe growth retardation of cTPx II mutant resulted in accumulation of G(1) phased cells. Based on kinetic properties of five isoforms, their subcellular localizations, and distinct physiology of each null mutant, we discussed the physiological functions of five types of TPx isoenzymes in yeast throughout the full growth cycle. PMID- 10681559 TI - p76(MDM2) inhibits the ability of p90(MDM2) to destabilize p53. AB - The mdm2 oncogene encodes p90(MDM2), which binds to and inactivates the p53 tumor suppressor protein. p90(MDM2) inhibits p53 by blocking the transcriptional activation domain of p53 as well as by stimulating its degradation. Recently, we showed that another product of the wild-type mdm2 gene, p76(MDM2), lacks the first 49 amino acids of p90(MDM2) and cannot bind p53. Here, we report that, like p90(MDM2), p76(MDM2) is expressed in both the nuclear and cytoplasmic compartments. Overexpression of p76(MDM2) antagonizes the ability of p90(MDM2) to stimulate the degradation of p53 and leads to an increase in the levels and activity of p53. Seven murine tissues express an alternatively spliced mdm2 mRNA that can encode p76(MDM2) but not p90(MDM2), as well as the normally spliced mdm2 mRNA that encodes both MDM2 proteins. All seven tissues express both MDM2 proteins. p90(MDM2) is much more abundant than p76(MDM2) in the testis, brain, heart, and kidney. However, in those tissues known to undergo p53-mediated apoptosis in response to gamma-irradiation, the thymus, spleen, and intestine, the levels of the MDM2 proteins are roughly equivalent. Our results indicate that the ratio of the two MDM2 proteins may regulate the response of tissues to DNA damage. PMID- 10681560 TI - Distinct roles for amino- and carboxyl-terminal sequences of SPRR1 protein in the formation of cross-linked envelopes of conducting airway epithelial cells. AB - The small proline-rich protein, SPRR1, is a marker gene whose expression in conducting airway epithelium is elevated under a variety of conditions that enhance squamous differentiation. The purpose of this study is to elucidate the nature of the SPRR1 sequence involved in cross-linked envelope formation in a tissue/cell type, such as conducting airway epithelium, that normally does not express squamous function except after injury or maintenance in culture. For this, a Flag-SPRR1 fusion protein expression system has been developed. Using the liposome-mediated gene transfer technique on passage 1 culture of human tracheobronchial epithelial (TBE) cells, the Flag-SPRR1 fusion protein can be expressed and detected immunologically by both anti-Flag and anti-SPRR1 antibodies. The incorporation of Flag-SPRR1 fusion protein into cross-linked envelopes can be demonstrated when transfected human passage 1 TBE cultures are treated with phorbol 12-myristate 13-acetate and high calcium (1.5 mM). By deletion and site-directed mutagenesis, two distinct roles of the amino- and carboxyl-terminal sequences of SPRR1 have been demonstrated. First, we demonstrated that the amino-terminal sequence of SPRR1 protein is required for the incorporation of the fusion protein into cross-linked envelopes, whereas a deletion on the carboxyl-terminal region or on the middle repetitive unit has no effect. Interestingly, insertion of a 24-amino acid peptide of monkey MUC2 repetitive sequence in the amino-terminus of SPRR1 protein had a stimulatory effect. Site-directed mutagenesis on the following amino acid residues, Lys(7), Gln(88), and Lys(89), which were found previously to participate in the cross linked envelope formation of keratinocytes, had no detrimental effect on the incorporation. However, mutations on Gln clusters, such as Gln(4)-Gln(6) and Gln(22)-Gln(25), had detrimental effects on the incorporation. These results suggest an amino-terminal sequence-dependent and multiple cross-linked sites for the incorporation of Flag-SPRR1 fusion protein into cross-linked envelopes of cultured human TBE cells. Second, we demonstrated that the carboxyl terminus of SPRR1 protein is required for a high level of Flag-fusion protein expression. A deletion in the carboxyl region or a mutation on the last lysine residue of the carboxyl end had a detrimental effect on the level of Flag-SPRR1 fusion protein expressed in transfected cells. In contrast, there was only a slight decrease in the level of expression if the amino-terminus was deleted. Interestingly, the efficiency for fusion protein to incorporate into cross-linked envelopes was elevated by the mutation at the carboxyl end. These results suggest distinct roles, perhaps coordinately, for both amino- and carboxyl-terminal sequences in the regulation of the life cycle of SPRR1 protein in cultured TBE cells. PMID- 10681561 TI - Epiregulin, a novel member of the epidermal growth factor family, is an autocrine growth factor in normal human keratinocytes. AB - Epiregulin is a new member of the epidermal growth factor (EGF) family purified from conditioned medium of NIH-3T3 clone T7. Some EGF family growth factors play essential roles in human keratinocytes in an autocrine manner. We show here that epiregulin is another autocrine growth factor for human keratinocytes. Epiregulin stimulated human keratinocyte proliferation under both subconfluent and confluent culture conditions in the absence of exogenous EGF family growth factors. Immunoprecipitation of [(35)S]methionine-labeled conditioned medium revealed a 5 kDa band corresponding to epiregulin. Northern blot analysis detected a 4. 8 kilobase transcript of epiregulin, and the addition of epiregulin up-regulated epiregulin mRNA synthesis. Furthermore, an anti-epiregulin blocking antibody reduced DNA synthesis by 25%. Epiregulin up-regulated the mRNA levels of heparin binding EGF-like growth factor (HB-EGF), amphiregulin, and TGF-alpha. In turn, the addition of EGF, HB-EGF, amphiregulin, and TGF-alpha increased epiregulin mRNA levels. These results demonstrate that epiregulin acts as an autocrine growth factor in human epidermal keratinocytes and is part of auto- and cross induction mechanisms involving HB-EGF, amphiregulin, and TGF-alpha. The mRNA expression profile resulting from induction of differentiation with high calcium and fetal calf serum revealed the differential expression of epiregulin, HB-EGF, amphiregulin, and TGF-alpha in keratinocytes. This indicates that these four growth factors have distinct, non-redundant biological functions. PMID- 10681562 TI - A dominant-negative peroxisome proliferator-activated receptor gamma (PPARgamma) mutant is a constitutive repressor and inhibits PPARgamma-mediated adipogenesis. AB - The nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) promotes adipocyte differentiation, exerts atherogenic and anti-inflammatory effects in monocyte/macrophages, and is believed to mediate the insulin sensitizing action of antidiabetic thiazolidinedione ligands. As no complete PPARgamma antagonists have been described hitherto, we have constructed a dominant-negative mutant receptor to inhibit wild-type PPARgamma action. Highly conserved hydrophobic and charged residues (Leu(468) and Glu(471)) in helix 12 of the ligand-binding domain were mutated to alanine. This compound PPARgamma mutant retains ligand and DNA binding, but exhibits markedly reduced transactivation due to impaired coactivator (cAMP-response element-binding protein-binding protein and steroid receptor coactivator-1) recruitment. Unexpectedly, the mutant receptor silences basal gene transcription, recruits corepressors (the silencing mediator of retinoid and thyroid receptors and the nuclear corepressor) more avidly than wild-type PPARgamma, and exhibits delayed ligand-dependent corepressor release. It is a powerful dominant-negative inhibitor of cotransfected wild-type receptor action. Furthermore, when expressed in primary human preadipocytes using a recombinant adenovirus, this PPARgamma mutant blocks thiazolidinedione-induced differentiation, providing direct evidence that PPARgamma mediates adipogenesis. Our observations suggest that, as in other mutant nuclear receptor contexts (acute promyelocytic leukemia, resistance to thyroid hormone), dominant-negative inhibition by PPARgamma is linked to aberrant corepressor interaction. Adenoviral expression of this mutant receptor is a valuable means to antagonize PPARgamma signaling. PMID- 10681563 TI - Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation. AB - The platelet integrin glycoprotein (GP) IIb/IIIa, which mediates platelet aggregation, has been the target for novel antiplatelet agents, the GPIIb/IIIa antagonists. Several GPIIb/IIIa antagonists have been developed based on the peptide RGDS present in adhesion proteins, including the principle ligand fibrinogen. The apoptosis enzyme, procaspase-3, contains an RGD-recognition sequence and is activated by RGDS. We examined the effects of RGDS and several GPIIb/IIIa antagonists on cell death and procaspase-3 activation in rat neonatal cardiomyocytes. These antagonists do not recognize rat integrins, yet RGDS, orbofiban, and xemilofiban induced dose-dependent apoptosis and procaspase-3 activation in cardiomyocytes over 72 h, particularly under hypoxic conditions. Scrambled peptide, the monoclonal antibody 7E3 or integrelin (a peptide containing a KGD sequence), had little or no effect. Immunoprecipitation of procaspase-3 followed by treatment with the compounds showed that procaspase-3 was activated directly by RGDS, orbofiban, xemilofiban, and by monoclonal 7E3 antibody, the latter demonstrating that compounds must enter cells to induce apoptosis through caspase activation. Integrelin had no effect. Binding studies with (3)H-SC52012B, a GPIIb/IIIa antagonist analogue of orbofiban, showed no specific binding to cardiomyocytes, but the radioligand accumulated intracellularly over 72 h. (3)H-SC52012B also bound directly to human recombinant caspase-3 (K(d), 59 +/- 2 nm), and this was prevented by orbofiban, xemilofiban, and the monoclonal 7E3 antibody but not by integrelin. Finally confocal microscopy showed that RGDS co-localized with caspase-3 inside the cell. These data show that RGDS and its mimetics induce cardiomyocyte apoptosis by direct activation of procaspase-3. PMID- 10681564 TI - The assembly factor Atp11p binds to the beta-subunit of the mitochondrial F(1) ATPase. AB - Atp11p is a protein of Saccharomyces cerevisiae required for the assembly of the F(1) component of the mitochondrial F(1)F(0)-ATP synthase. This study presents evidence that Atp11p binds selectively to the beta-subunit of F(1). Under conditions in which avidin-Sepharose beads specifically adsorbed biotinylated Atp11p from yeast mitochondrial extracts, the F(1) beta-subunit coprecipitated with the tagged Atp11p protein. Binding interactions between Atp11p and the entire beta-subunit of F(1) or fragments of the beta-subunit were also revealed by a yeast two-hybrid screen: Atp11p bound to a region of the nucleotide-binding domain of the beta-subunit located between Gly(114) and Leu(318). Certain elements of this sequence that would be accessible to Atp11p in the free beta subunit make contact with adjacent alpha-subunits in the assembled enzyme. This observation suggests that the alpha-subunits may exchange for bound Atp11p during the process of F(1) assembly. PMID- 10681565 TI - Occludin modulates transepithelial migration of neutrophils. AB - Neutrophils cross epithelial sheets to reach inflamed mucosal surfaces by migrating along the paracellular route. To avoid breakdown of the epithelial barrier, this process requires coordinated opening and closing of tight junctions, the most apical intercellular junctions in epithelia. To determine the function of epithelial tight junction proteins in this process, we analyzed neutrophil migration across monolayers formed by stably transfected epithelial cells expressing wild-type and mutant occludin, a membrane protein of tight junctions with four transmembrane domains and both termini in the cytosol. We found that expression of mutants with a modified N-terminal cytoplasmic domain up regulated migration, whereas deletion of the C-terminal cytoplasmic domain did not have an effect. The N-terminal cytosolic domain was also found to be important for the linear arrangement of occludin within tight junctions but not for the permeability barrier. Moreover, expression of mutant occludin bearing a mutation in one of the two extracellular domains inhibited neutrophil migration. The effects of transfected occludin mutants on neutrophil migration did not correlate with their effects on selective paracellular permeability and transepithelial electrical resistance. Hence, specific domains and functional properties of occludin modulate transepithelial migration of neutrophils. PMID- 10681566 TI - Discoidin domain receptor 1 is activated independently of beta(1) integrin. AB - Various types of collagen have been identified as potential ligands for the two mammalian discoidin domain receptor (DDR) tyrosine kinases, DDR1 and DDR2. It is presently unclear whether collagen-induced DDR receptor activation, which occurs with very slow kinetics, involves additional proteins with kinase activity or membrane-anchored proteins serving as coreceptors. In particular, the role of the collagen-binding integrins alpha(1)beta(1) or alpha(2)beta(1) in the DDR activation process is undefined. Here, we provide three lines of evidence suggesting that DDR1 signaling is distinct from integrin activation. First we demonstrate that the enzymatic activity of DDR1 is essential for receptor tyrosine phosphorylation. Collagen-induced DDR receptor autophosphorylation can be blocked either by a dominant negative mutant or by a preparation of recombinant extracellular domain. Second, we show DDR1 signals independent of the epidermal growth factor (EGF) receptor. In cells that endogenously express both DDR1 and the EGF receptor, stimulation with EGF does not induce DDR activation. Third, we detected full DDR1 activation after collagen stimulation in cells that have been treated with blocking antibodies for alpha(2)beta(1) integrin or in cells with a targeted deletion of the beta(1) integrin gene. Finally, we show that overexpression of dominant negative DDR1 in the myoblast cell line C2C12 blocks cellular differentiation and the formation of myofibers. PMID- 10681567 TI - Structures, enzymatic properties, and expression of novel human and mouse secretory phospholipase A(2)s. AB - Mammalian secretory phospholipase A(2)s (sPLA(2)s) form a family of structurally related enzymes that are involved in a variety of physiological and pathological processes via the release of arachidonic acid from membrane phospholipids or the binding to specific membrane receptors. Here, we report the cloning and characterization of a novel sPLA(2) that is the sixth isoform of the sPLA(2) family found in humans. The novel human mature sPLA(2) consists of 123 amino acids (M(r) = 14,000) and is most similar to group IIA sPLA(2) (sPLA(2)-IIA) with respect to the number and positions of cysteine residues as well as overall identity (51%). Therefore, this novel sPLA(2) should be categorized into group II and called group IIE (sPLA(2)-IIE) following the recently identified group IID sPLA(2) (sPLA(2)-IID). The enzymatic properties of recombinant human sPLA(2)-IIE were almost identical to those of sPLA(2)-IIA and IID in terms of Ca(2+) requirement, optimal pH, substrate specificity, as well as high susceptibility to the sPLA(2) inhibitor indoxam. Along with the biochemical properties of proteins, genetic and evolutional similarities were also observed among these three types of group II sPLA(2)s as to the chromosomal location of the human gene (1p36) and the exon/intron organization. The expression of sPLA(2)-IIE transcripts in humans was restricted to the brain, heart, lung, and placenta in contrast to broad expression profiles for sPLA(2)-IIA and -IID. In sPLA(2)-IIA-deficient mice, the expression of sPLA(2)-IIE was markedly enhanced in the lung and small intestine upon endotoxin challenge, which contrasted with the reduced expression of sPLA(2) IID mRNA. In situ hybridization analysis revealed elevation of sPLA(2)-IIE mRNA at alveolar macrophage-like cells in the lung of endotoxin-treated mice. These findings suggest a distinct functional role of novel sPLA(2)-IIE in the progression of inflammatory processes. PMID- 10681568 TI - Acetyl-CoA synthetase from the amitochondriate eukaryote Giardia lamblia belongs to the newly recognized superfamily of acyl-CoA synthetases (Nucleoside diphosphate-forming). AB - The gene coding for the acetyl-CoA synthetase (ADP-forming) from the amitochondriate eukaryote Giardia lamblia has been expressed in Escherichia coli. The recombinant enzyme exhibited the same substrate specificity as the native enzyme, utilizing acetyl-CoA and adenine nucleotides as preferred substrates and less efficiently, propionyl- and succinyl-CoA. N- and C-terminal parts of the G. lamblia acetyl-CoA synthetase sequence were found to be homologous to the alpha- and beta-subunits, respectively, of succinyl-CoA synthetase. Sequence analysis of homologous enzymes from various bacteria, archaea, and the eukaryote, Plasmodium falciparum, identified conserved features in their organization, which allowed us to delineate a new superfamily of acyl-CoA synthetases (nucleoside diphosphate forming) and its signature motifs. The representatives of this new superfamily of thiokinases vary in their domain arrangement, some consisting of separate alpha- and beta-subunits and others comprising fusion proteins in alpha-beta or beta alpha orientation. The presence of homologs of acetyl-CoA synthetase (ADP forming) in such human pathogens as G. lamblia, Yersinia pestis, Bordetella pertussis, Pseudomonas aeruginosa, Vibrio cholerae, Salmonella typhi, Porphyromonas gingivalis, and the malaria agent P. falciparum suggests that they might be used as potential drug targets. PMID- 10681569 TI - Hormonal regulation of the phosphoenolpyruvate carboxykinase gene. Role of specific CCAAT/enhancer-binding protein isoforms. AB - The CCAAT/enhancer-binding protein alpha (C/EBP) is a transcription factor that trans-activates a number of metabolically important genes. Previous work has demonstrated that C/EBPalpha and C/EBPbeta have the potential to mediate the cAMP responsiveness of phosphoenolpyruvate carboxykinase (PEPCK) in liver cells. However, these studies used GAL4 fusion proteins and artificial promoter-reporter gene vectors in transfection experiments; as a result, these studies only indicated that both isoforms had the potential to mediate the hormonal response and not which isoform actually participated in vivo. To address this issue, we produced hepatoma cell lines that stably expressed either a dominant negative inhibitor or antisense RNA for these two main liver C/EBP isoforms. Inhibition of all C/EBP isoforms via expression of the dominant negative protein eliminated cAMP responsiveness, and reduced glucocorticoid responsiveness, of the endogenous PEPCK gene in hepatoma cells. Antisense directed against C/EBPalpha mRNA, which reduced C/EBPalpha protein levels by nearly 80%, also significantly reduced the cAMP responsiveness of the endogenous PEPCK promoter, whereas antisense directed against C/EBPbeta was without effect. There was no major alteration in cAMP signaling in the C/EBPalpha antisense cells, as cAMP induction of the C/EBPbeta gene was similar to that in wild-type H4IIE cells. These data suggest that the alpha-isoform of C/EBP is specifically utilized for mediating the cAMP responsiveness of the PEPCK gene. PMID- 10681570 TI - Sites of action of protein kinase C and phosphatidylinositol 3-kinase are distinct in oxidized low density lipoprotein-induced macrophage proliferation. AB - Oxidized low density lipoprotein (Ox-LDL) can induce macrophage proliferation in vitro. To explore the mechanisms involved in this process, we reported that activation of protein kinase C (PKC) is involved in its signaling pathway (Matsumura, T., Sakai, M., Kobori, S., Biwa, T., Takemura, T., Matsuda, H., Hakamata, H., Horiuchi, S., and Shichiri, M. (1997) Arterioscler. Thromb. Vasc. Biol. 17, 3013-3020) and that expression of granulocyte/macrophage colony stimulating factor (GM-CSF) and its subsequent release in the culture medium are important (Biwa, T., Hakamata, H., Sakai, M., Miyazaki, A., Suzuki, H., Kodama, T., Shichiri, M., and Horiuchi, S. (1998) J. Biol. Chem. 273, 28305-28313). However, a recent study also demonstrated the involvement of phosphatidylinositol 3-kinase (PI3K) in this process. In the present study, we investigated the role of PKC and PI3K in Ox-LDL-induced macrophage proliferation. Ox-LDL-induced macrophage proliferation was inhibited by 90% by a PKC inhibitor, calphostin C, and 50% by a PI3K inhibitor, wortmannin. Ox-LDL-induced expression of GM-CSF and its subsequent release were inhibited by calphostin C but not by wortmannin, whereas recombinant GM-CSF-induced macrophage proliferation was inhibited by wortmannin by 50% but not by calphostin C. Ox-LDL activated PI3K at two time points (10 min and 4 h), and the activation at the second but not first point was significantly inhibited by calphostin C and anti-GM-CSF antibody. Our results suggest that PKC plays a role upstream in the signaling pathway to GM-CSF induction, whereas PI3K is involved, at least in part, downstream in the signaling pathway after GM-CSF induction. PMID- 10681571 TI - Isolation of the ace1 gene encoding a Cys(2)-His(2) transcription factor involved in regulation of activity of the cellulase promoter cbh1 of Trichoderma reesei. AB - A genetic selection method was developed for the cloning of positive-acting transcriptional regulatory genes in Saccharomyces cerevisiae. The method was applied for the isolation of activators of Trichoderma reesei (Hypocrea jecorina) cellulase genes. Activator genes were isolated from a T. reesei expression cDNA library on the basis of the ability of their translation products to activate transcription from the full-length T. reesei cbh1 promoter coupled to the S. cerevisiae HIS3 gene and to support the growth of the yeast colonies in the absence of histidine. Among the clones obtained was the ace1 gene encoding a novel polypeptide, ACEI, that contains three zinc finger motifs of Cys(2)-His(2) type. Possible ACEI homologues were found among expressed sequence tags of Aspergillus and Neurospora. The ability of ACEI to bind to the cbh1 promoter was further confirmed in the yeast one-hybrid system. In vitro binding and gel mobility shift assays revealed several binding sites for the ACEI protein in the cbh1 promoter. Disruption of the ace1 gene in T. reesei resulted in retarded growth of the fungus on a cellulose-containing medium, on which cellulases are normally highly expressed. PMID- 10681572 TI - Identical or overlapping sequences in the primary structure of human alpha(2) macroglobulin are responsible for the binding of nerve growth factor-beta, platelet-derived growth factor-BB, and transforming growth factor-beta. AB - alpha(2)-Macroglobulin (alpha(2)M) functions as a proteinase inhibitor and as a carrier of diverse growth factors. In this study, we localized binding sites for platelet-derived growth factor-BB (PDGF-BB) and nerve growth factor-beta (NGF beta) to a linear sequence in the 180-kDa human alpha(2)M subunit which includes amino acids 591-774. A glutathione S-transferase fusion protein containing amino acids 591-774 (FP3) bound PDGF-BB and NGF-beta in ligand blotting assays whereas five other fusion proteins, which collectively include amino acids 99-590 and 775 1451 did not. The K(D) values for PDGF-BB and NGF-beta binding to immobilized FP3 were 300 +/- 40 and 180 +/- 30 nM, respectively; these values were comparable with those determined using methylamine-modified alpha(2)M, suggesting that higher-order alpha(2)M structure is not necessary for PDGF-BB and NGF-beta binding. PDGF-BB and NGF-beta blocked the binding of transforming growth factor beta1 (TGF-beta1) to FP3. Furthermore, murinoglobulin, which is the only known member of the alpha-macroglobulin family that does not bind TGF-beta, also failed to bind PDGF-BB and NGF-beta. These results support the hypothesis that either a single linear sequence in human alpha(2)M or overlapping sequences are responsible for the binding of TGF-beta, PDGF-BB, and NGF-beta, even though there is minimal sequence identity between these three growth factors. FP3 blocked the binding of PDGF-BB to a purified chimeric protein, in which the extracellular domain of the PDGF beta receptor was fused to the IgG(1) Fc domain, and to PDGF receptors on NIH 3T3 cells. Thus, FP3 may inhibit the activity of PDGF-BB. PMID- 10681573 TI - GD3 synthase gene expression in PC12 cells results in the continuous activation of TrkA and ERK1/2 and enhanced proliferation. AB - A rat pheochromocytoma cell line (PC12) transfected with ganglioside GD3 synthase gene showed a marked change in the ganglioside profile and enhanced proliferation and no response of neurite extension to nerve growth factor (NGF) stimulation. In these transfectant cells, a continuous phosphorylation of TrkA and the activation of ERK1/2 without NGF treatment were observed. Proliferation inhibition experiments with kinase inhibitors such as herbimycin A, K-252a, and PD98059 revealed that the enhanced proliferation was actually due to the activation of the Ras/MEK/ERK pathway. A TrkA dimer was detected in the GD3 synthase transfectant cells regardless of NGF treatment by cross-linking and immunoblotting. The increased expression of GD1b and GT1b in these transfectant cells might induce the conformational change of TrkA to form a dimer and to be activated continuously. These results may indicate regulatory roles of gangliosides in cell proliferation under physiological and malignant processes. PMID- 10681574 TI - Partitioning of serpin-proteinase reactions between stable inhibition and substrate cleavage is regulated by the rate of serpin reactive center loop insertion into beta-sheet A. AB - The serpin family of serine proteinase inhibitors is a mechanistically unique class of naturally occurring proteinase inhibitors that trap target enzymes as stable covalent acyl-enzyme complexes. This mechanism appears to require both cleavage of the serpin reactive center loop (RCL) by the proteinase and a significant conformational change in the serpin structure involving rapid insertion of the RCL into the center of an existing beta-sheet, serpin beta-sheet A. The present study demonstrates that partitioning between inhibitor and substrate modes of reaction can be altered by varying either the rates of RCL insertion or deacylation using a library of serpin RCL mutants substituted in the critical P(14) hinge residue and three different proteinases. We further correlate the changes in partitioning with the actual rates of RCL insertion for several of the variants upon reaction with the different proteinases as determined by fluorescence spectroscopy of specific RCL-labeled inhibitor mutants. These data demonstrate that the serpin mechanism follows a branched pathway, and that the formation of a stable inhibited complex is dependent upon both the rate of the RCL conformational change and the rate of enzyme deacylation. PMID- 10681575 TI - The itinerary of a vesicle component, Aut7p/Cvt5p, terminates in the yeast vacuole via the autophagy/Cvt pathways. AB - Aminopeptidase I (API) is delivered to the yeast vacuole by one of two alternative pathways, cytoplasm to vacuole targeting (Cvt) or autophagy, depending on nutrient conditions. Genetic, morphological, and biochemical studies indicate that the two pathways share many of the same molecular components. The Cvt pathway functions during vegetative growth, while autophagy is induced during starvation. Both pathways involve the formation of cytosolic vesicles that fuse with the vacuole. In either case, the mechanism of vesicle formation is not known. Autophagic uptake displays a greater capacity for cytosolic protein sequestration. This suggests the involvement of an inducible protein(s) that allows the vesicle-forming machinery to adapt to the increased degradative needs of the cell. We have analyzed the biosynthesis of Aut7p, a protein required for both pathways. We find Aut7p expression is induced by nitrogen starvation. Aut7p is degraded by a process dependent on both proteinase A and Cvt/autophagy components. Protease accessibility assays demonstrate that Aut7p is located within vesicles in strains defective in vesicle delivery or breakdown. Finally, the aut7/cvt5 mutant accumulates precursor API at a stage prior to vesicle completion. These data suggest that Aut7p is induced during autophagy and delivered to the vacuole together with precursor API by Cvt/autophagic vesicles. PMID- 10681576 TI - Limitation in electron transfer in photosystem I donor side mutants of Chlamydomonas reinhardtii. Lethal photo-oxidative damage in high light is overcome in a suppressor strain deficient in the assembly of the light harvesting complex. AB - Strains of Chlamydomonas reinhardtii lacking the PsaF gene or containing the mutation K23Q within the N-terminal part of PsaF are sensitive to high light (>400 microE m(-2) s(-1)) under aerobic conditions. In vitro experiments indicate that the sensitivity to high light of the isolated photosystem I (PSI) complex from wild type and from PsaF mutants is similar. In vivo measurements of photochemical quenching and oxygen evolution show that impairment of the donor side of PSI in the PsaF mutants leads to a diminished linear electron transfer and/or a decrease of photosystem II (PSII) activity in high light. Thermoluminescence measurements indicate that the PSII reaction center is directly affected under photo-oxidative stress when the rate of electron transfer becomes limiting in the PsaF-deficient strain and in the PsaF mutant K23Q. We have isolated a high light-resistant PsaF-deficient suppressor strain that has a high chlorophyll a/b ratio and is affected in the assembly of light-harvesting complex. These results indicate that under high light a functionally intact donor side of PSI is essential for protection of C. reinhardtii against photo-oxidative damage when the photosystems are properly connected to their light-harvesting antennae. PMID- 10681577 TI - The insulin-like growth factors (IGFs) I and II bind to articular cartilage via the IGF-binding proteins. AB - Bovine articular cartilage discs (3 mm diameter x 400 micrometer thick) were equilibrated in buffer containing (125)I-insulin-like growth factor (IGF)-I (4 degrees C) +/- unlabeled IGF-I or IGF-II. Competition for binding to cartilage discs by each unlabeled IGF was concentration-dependent, with ED(50) values for inhibition of (125)I-IGF-I binding of 11 and 10 nM for IGF-I and -II, respectively, and saturation by 50 nM. By contrast, an analog of IGF-I with very low affinity for the insulin-like growth factor-binding proteins (IGF-BPs), des (1-3)-IGF-I, was not competitive with (125)I-IGF-I for cartilage binding even at 100-400 nM. Binding of the (125)I-labeled IGF-II isoform to cartilage was competed for by unlabeled IGF-I or -II, with ED(50)s of 160 and 8 nM, respectively. This probably reflected the differential affinities of the endogenous IGF-BPs (IGF-BP-6 and -2) for IGF-II/IGF-I. Transport of (125)I-IGF-I was also measured in an apparatus that allows diffusion only across the discs (400 micrometer), by addition to one side and continuous monitoring of efflux on the other side. The time lag for transport of (125)I-IGF was 266 min, an order of magnitude longer than the theoretical prediction for free diffusion in the matrix. (125)I-IGF-I transport then reached a steady state rate (% efflux of total added (125)I-IGF/unit time), which was subsequently accelerated approximately 2-fold by addition of an excess of unlabeled IGF-I. Taken together, these results indicate that IGF binding to cartilage, mostly through the IGF-BPs, regulates the transport of IGFs in articular cartilage, probably contributing to the control of their paracrine activities. PMID- 10681578 TI - Proteinase inhibitor 9, an inhibitor of granzyme B-mediated apoptosis, is a primary estrogen-inducible gene in human liver cells. AB - Although liver is an estrogen target tissue, the number of hepatic genes known to be directly induced by estrogen is very small. We identified proteinase inhibitor 9, or PI-9, as being rapidly and strongly induced by estrogen in an estrogen receptor-positive human liver cell line (HepG2-ER7). Since PI-9 mRNA was also induced by estrogen in a human liver biopsy sample, PI-9 is a genuine estrogen regulated human gene. PI-9 is a potent inhibitor of granzyme B and of granzyme B mediated apoptosis. Estrogens induced PI-9 mRNA within 2 h, PI-9 mRNA levels reached a plateau of 30-40-fold induction in 4 h, and induction was not blocked by cycloheximide, indicating that induction of PI-9 mRNA is a primary response. The antiestrogen trans-hydroxytamoxifen was a partial agonist for PI-9 mRNA induction, whereas the antiestrogen ICI 182, 780 was a pure antagonist. Western blot analysis showed that estrogen strongly increases PI-9 protein levels. Inhibition of transcription with actinomycin D resulted in identical rates of PI 9 mRNA decay in the presence and absence of estrogen. We isolated genomic clones containing the PI-9 promoter region, identified a putative transcription start site, and carried out transient transfections of PI-9-luciferase reporter gene constructs. The estrogen, moxestrol, elicited a robust induction from the PI-9 luciferase reporter. Mutational inactivation of three potential imperfect estrogen response elements in the PI-9 5'-flanking region had no effect on moxestrol estrogen receptor induction. PMID- 10681579 TI - Dissecting the interactions between NTF2, RanGDP, and the nucleoporin XFXFG repeats. AB - We have used a range of complementary biochemical and biophysical methods to investigate the interactions between nuclear transport factor 2 (NTF2), the Ras family GTPase Ran, and XFXFG nucleoporin repeats that are crucial for nuclear trafficking. Microcalorimetry, microtiter plate binding, and fluorescence quenching in solution are all consistent with the binding constant for the NTF2 RanGDP interaction being in the 100 nM range, whereas the interaction between NTF2 and XFXFG repeat-containing nucleoporins such as Nsp1p is in the 1 microM range. Although the accumulation of NTF2 at the nuclear envelope is enhanced by RanGDP, we show that Ran binding does not alter the affinity of NTF2 for nucleoporins nor does the binding of nucleoporins alter the affinity of NTF2 for RanGDP. These results indicate that, instead, Ran increases the binding of NTF2 to nucleoporins by another mechanism, most probably by Ran itself binding to nucleoporins and NTF2 binding to this nuclear pore-associated Ran. PMID- 10681580 TI - A novel conformation of the herpes simplex virus origin of DNA replication recognized by the origin binding protein. AB - The Herpes simplex virus type I origin binding protein (OBP) is a sequence specific DNA-binding protein and a dimeric DNA helicase encoded by the UL9 gene. It is required for the activation of the viral origin of DNA replication oriS. Here we demonstrate that the linear double-stranded form of oriS can be converted by heat treatment to a stable novel conformation referred to as oriS*. Studies using S1 nuclease suggest that oriS* consists of a central hairpin with an AT rich sequence in the loop. Single-stranded oligonucleotides corresponding to the upper strand of oriS can adopt the same structure. OBP forms a stable complex with oriS*. We have identified structural features of oriS* recognized by OBP. The central oriS palindrome as well as sequences at the 5' side of the oriS palindrome were required for complex formation. Importantly, we found that mutations that have been shown to reduce oriS-dependent DNA replication also reduce the formation of the OBP-oriS* complex. We suggest that oriS* serves as an intermediate in the initiation of DNA replication providing the initiator protein with structural information for a selective and efficient assembly of the viral replication machinery. PMID- 10681581 TI - A minimized human integrin alpha(5)beta(1) that retains ligand recognition. AB - Two isolated recombinant fragments from human integrin alpha(5)beta(1) encompassing the FG-GAP repeats III to VII of alpha(5) and the insertion-type domain from beta(1), respectively, are structurally well defined in solution, based on CD evidence. Divalent cation binding induces a conformational adaptation that is achieved by Ca(2+) or Mg(2+) (or Mn(2+)) with alpha(5) and only by Mg(2+) (or Mn(2+)) with beta(1). Mn(2+) bound to beta(1) is highly hydrated ( approximately 3 water molecules), based on water NMR relaxation, in agreement with a metal ion-dependent adhesion site-type metal coordination. Each fragment saturated with Mg(2+) (or Mn(2+)) binds a recombinant fibronectin ligand in an RGD-dependent manner. A conformational rearrangement is induced on the fibronectin ligand upon binding to the alpha(5), but not to the beta(1) fragment, based on CD. Ligand binding results in metal ion displacement from beta(1). Both alpha(5) and beta(1) fragments form a stable heterodimer (alpha(5)beta(1) mini integrin) that retains ligand recognition to form a 1:1:1 ternary complex, in the presence of Mg(2+), and induces a specific conformational adaptation of the fibronectin ligand. A two-site model for RGD binding to both alpha and beta integrin components is inferred from our data using low molecular weight RGD mimetics. PMID- 10681582 TI - Association of human origin recognition complex 1 with chromatin DNA and nuclease resistant nuclear structures. AB - An origin recognition complex (ORC) consisting of six polypeptides has been identified as a DNA replication origin-binding factor in Saccharomyces cerevisiae. Homologues of ORC subunits have been discovered among eukaryotes, and we have prepared monoclonal antibodies against a human homologue of ORC1 (hORC1) to study its localization in human cells. It was thus found to associate with nuclei throughout the cell cycle and to be resistant to nonionic detergent treatment, in contrast to MCM proteins, which are other replication factors, the association of which with nuclei is clearly dependent on the phase of the cell cycle. A characteristic feature of hORC1 is dissociation by NaCl in a narrow concentration range around 0.25 M, suggesting interaction with some specific partner(s) in nuclei. Nuclease treatment experiments and UV cross-linking experiments further indicated interaction with both nuclease-resistant nuclear structures and chromatin DNA. Although its DNA binding was unaffected, some variation in the cell cycle was apparent, the association with nuclear structures being less stable in the M phase. Interestingly, the less stable association occurred concomitantly with hyperphosphorylation of hORC1, suggesting that this hyperphosphorylation may be involved in M phase changes. PMID- 10681583 TI - Differential effect of Rac and Cdc42 on p38 kinase activity and cell cycle progression of nonadherent primary mouse fibroblasts. AB - The Rho GTPases play an important role in transducing signals linking plasma membrane receptors to the organization of the cytoskeleton and also regulate gene transcription. Here, we show that expression of constitutively active Ras or Cdc42, but not RhoA, RhoG, and Rac1, is sufficient to cause anchorage-independent cell cycle progression of mouse embryonic fibroblasts. However, in anchorage free conditions, whereas activation of either Cdc42 or Ras results in cyclin A transcription and cell cycle progression, Cdc42 is not required for Ras-mediated cyclin A induction, and the two proteins act in a synergistic manner in this process. Surprisingly, the ability of Cdc42 to induce p38 MAPK activity in suspended mouse embryonic fibroblast was impaired. Moreover, inhibition of p38 activity allowed Rac1 to induce anchorage-independent cyclin A transcription, indicating that p38 MAPK has an inhibitory function on cell cycle progression of primary fibroblasts. Finally, a Rac mutant, which is unable to induce lamellipodia and focal complex formation, promoted cyclin A transcription in the presence of SB203580, suggesting that the organization of the cytoskeleton is not required for anchorage-independent proliferation. This demonstrates a novel function for Cdc42, distinct from that of Rac1, in the control of cell proliferation. PMID- 10681584 TI - Repressed expression of the human xanthine oxidoreductase gene. E-box and TATA like elements restrict ground state transcriptional activity. AB - Studies were initiated to address the basis for the low xanthine oxidoreductase (XOR) activity in humans relative to nonprimate mammalian species. The expression of the XOR in humans is strikingly lower than in mice, and both transcription rates and core promoter activity of the gene are repressed. Analysis of human XOR promoter activity in hepatocytes and vascular endothelial cells showed that the region from -258 to -1 contains both repressor and activator binding regions regulating core promoter activity. The region between -138 and -1 is necessary and sufficient for initiating, and the region between -258 and -228 is critical for restricting core promoter activity. Within the latter region, site-directed mutations identified a consensus sequence "acacaggtgtgg" (-242 to -230) that contains an E-box that binds a repressor. In addition, the TATA-like element is also required to restrict promoter activity and TFIID binds to this site. The results demonstrate that both an E-box and TATA-like element are required to restrict gene activity. A model is proposed to account for human XOR regulation. PMID- 10681585 TI - Induction of light chain replacement in human plasma cells by caffeine is independent from both the upregulation of RAG protein expression and germ line transcription. AB - When some human plasma cell lines are cultured with concanavalin A, the original light chain is replaced with another light chain which results from secondary VJ recombination (light chain shifting). We examined various intracellular factors involved in the induction of light chain shifting. Light chain shifting can be induced upon treatment with agents with phosphatase inhibitory activity such as caffeine and okadaic acid. Although the plasma cells used express both RAG-1 and RAG-2, the expression level of these proteins was not affected by caffeine or okadaic acid. Transcription of the germ line locus, which correlates to the locus activation for rearrangement, is also not influenced by phosphatase inhibition. However, the amount of signal broken-ended DNA intermediates generated during V(D)J rearrangement was shown to increase upon caffeine or okadaic acid treatment. The inhibitory activity of caffeine on phosphatase was the same as okadaic acid. However, caffeine exhibited much higher activity for VJ coding joint formation than okadaic acid. Therefore, although phosphatase inhibition might act, in part, on a mechanism by which V(D)J recombinase activity is regulated within the human plasma cells, other factor(s) are probably also involved in the process. PMID- 10681586 TI - Isolation, structure, synthesis, and activity of a new member of the calcitonin gene-related peptide family from frog skin and molecular cloning of its precursor. AB - Calcitonin gene-related peptide has been extracted from the skin exudate of a single living specimen of the frog Phyllomedusa bicolor and purified to homogeneity by a two-step protocol. A total volume of 250 microl of exudate yielded 380 microg of purified peptide. Mass spectrometric analysis and gas phase sequencing of the purified peptide as well as chemical synthesis and cDNA analysis were consistent with the structure SCDTSTCATQRLADFLSRSGGIGSPDFVPTDVSANSF amide and the presence of a disulfide bridge linking Cys(2) and Cys(7). The skin peptide, named skin calcitonin gene-related peptide, differs significantly from all other members of the calcitonin gene-related peptide family of peptides at nine positions but binds with high affinity to calcitonin gene-related peptide receptors in the rat brain and acts as an agonist in the rat vas deferens bioassay with potencies equal to those of human CGRP. Reverse transcriptase polymerase chain reaction coupled with cDNA cloning and sequencing demonstrated that skin calcitonin gene-related peptide isolated in the skin is identical to that present in the frog's central and enteric nervous systems. These data, which indicate for the first time the existence of calcitonin gene-related peptide in the frog skin, add further support to the brain-skin-gut triangle hypothesis as a useful tool in the identification and/or isolation of mammalian peptides that are present in the brain and other tissues in only minute quantities. PMID- 10681587 TI - Subunit interactions in yeast transcription/repair factor TFIIH. Requirement for Tfb3 subunit in nucleotide excision repair. AB - A yeast strain harboring a temperature-sensitive allele of TFB3 (tfb3(ts)), the 38-kDa subunit of the RNA polymerase II transcription/nucleotide excision repair factor TFIIH, was found to be sensitive to ultraviolet (UV) radiation and defective for nucleotide excision repair in vitro. Interestingly, tfb3(ts) failed to grow on medium containing caffeine. A comprehensive pairwise two-hybrid analysis between yeast TFIIH subunits identified novel interactions between Rad3 and Tfb3, Tfb4 and Ssl1, as well as Ssl2 and Tfb2. These interactions have facilitated a more complete model of the structure of TFIIH and the nucleotide excision repairosome. PMID- 10681588 TI - Identification and characterization of basal and cyclic AMP response elements in the promoter of the rat GTP cyclohydrolase I gene. AB - 5812 base pairs of rat GTP cyclohydrolase I (GTPCH) 5'-flanking region were cloned and sequenced, and the transcription start site was determined for the gene in rat liver. Progressive deletion analysis using transient transfection assays of luciferase reporter constructs defined the core promoter as a highly conserved 142-base pair GC-rich sequence upstream from the cap site. DNase I footprint analysis of this region revealed (5' --> 3') a Sp1/GC box, a noncanonical cAMP-response element (CRE), a CCAAT-box, and an E-box. Transcription from the core promoter in PC12 but not C6 or Rat2 cells was enhanced by incubation with 8-bromo-cyclic AMP. Mutagenesis showed that both the CRE and CCAAT-box independently contribute to basal and cAMP-dependent activity. The combined CRE and CCAAT-box cassette was also found to enhance basal transcription and confer cAMP sensitivity on a heterologous minimal promoter. The addition of the Sp1/GC box sequence to this minimal promoter construct inhibited basal transcription without affecting the cAMP response. EMSA showed that nuclear proteins from PC12 but not C6 or Rat2 cells bind the CRE as a complex containing activating transcription factor (ATF)-4 and CCAAT enhancer-binding protein beta, while both PC12 and C6 cell nuclear extracts were recruited by the CCAAT-box as a complex containing nuclear factor Y. Overexpression of ATF-4 in PC12 cells was found to transactivate the GTPCH promoter response to cAMP. These studies suggest that the elements required for cell type-specific cAMP-dependent enhancement of gene transcription are located along the GTPCH core promoter and include the CRE and adjacent CCAAT-box and the proteins ATF-4, CCAAT enhancer-binding protein beta, and nuclear factor Y. PMID- 10681589 TI - Structural analysis of alpha-enolase. Mapping the functional domains involved in down-regulation of the c-myc protooncogene. AB - Myc-binding protein-1 (MBP-1) is a 37-kDa protein with sequence homology to the 3' portion of the alpha-enolase gene. alpha-Enolase is a 48-kDa protein, which plays a critical role in the glycolytic pathway. MBP-1 binds to the c-myc P2 promoter and down-regulates c-myc expression. We have investigated the role of alpha-enolase in regulation of the c-myc protooncogene. RNase protection assay shows that alpha-enolase is transcribed into a single RNA species in HeLa cells. A start codon, 400 base pairs downstream of the alpha-enolase ATG, corresponds to the MBP-1 ATG, suggesting that MBP-1 is an alternative translation initiation product of the alpha-enolase RNA. Domain mapping was performed using constructs containing truncations of the alpha-enolase gene. In vitro binding to the c-myc gene was abolished after deletion of the N-terminal portion of alpha-enolase. In order to determine the relationship between DNA binding activity and transcription inhibition, we performed co-transfection assays in HeLa cells. These studies confirmed that an N-terminal deletion of alpha-enolase is unable to down-regulate c-myc promoter activity. Our data suggest that alpha-enolase plays an important role in regulation of c-myc promoter activity in the form of an alternative translation product MBP-1, which is distinct from its role as a glycolytic enzyme. PMID- 10681590 TI - ZAP-70 is essential for the T cell antigen receptor-induced plasma membrane targeting of SOS and Vav in T cells. AB - Translocation of the SOS and Vav GDP/GTP exchange factors proximal to Ras and Rac GTPases localized in the plasma membrane glycolipid-enriched microdomains is a pivotal step required for T cell antigen receptor-induced T cell activation. Here we demonstrate that the T cell antigen receptor zeta-chain-associated ZAP-70 kinase and T cell antigen receptor zeta-chain immunoreceptor tyrosine-based activation motifs are essential for the membrane recruitment of SOS and Vav. Plasma membrane targeting of SOS or Vav begins with the assembly of ZAP-70 with Grb-2 and SOS. The subsequent tyrosine phosphorylation of LAT (linker for activation of T cell) by ZAP-70 leads to a shift in equilibrium from the ZAP 70.Grb-2.SOS(Vav) complex to the (Vav)SOS.Grb-2.LAT complex. This shift results in the targeting of SOS and Vav into glycolipid-enriched microdomains and initiation of the Ras and Rac signaling cascades involved in T cell activation, proliferation, and cytokine production. PMID- 10681591 TI - Differential mechanisms of nuclear receptor regulation by receptor-associated coactivator 3. AB - Steroid and nuclear receptor coactivators (NCoAs) have been implicated in the regulation of nuclear receptor function by enhancing ligand-dependent transcriptional activation of target gene expression. We have previously isolated receptor-associated coactivator 3 (RAC3), which belongs to the steroid receptor coactivator family. In this study, we investigated the differential mechanisms by which RAC3 interacts with and modulates the transcriptional activity of different nuclear receptors. We found that the vitamin D receptor (VDR) and estrogen receptor beta interact with different alpha-helical LXXLL motifs of RAC3. Peptides corresponding to these motifs have diverse affinities for the VDR and estrogen receptor beta, and mutation of specific motifs differentially impairs the ability of RAC3 to interact with these receptors in vitro. Consequently, these mutations inhibit the enhancement of transcriptional activation by these receptors in vivo. Furthermore, we found that the activation function-2 (AF-2) domain of the retinoid X receptor interferes with RAC3 binding to a DNA-bound VDR/retinoid X receptor (RXR) heterodimer, whereas the VDR AF-2 domain is required for this interaction. These results suggest a receptor-specific binding preference for the different LXXLL motifs of RAC3, which may provide flexibility for RAC3 to differentially regulate the function of different nuclear receptors. PMID- 10681592 TI - SHP2 association with VE-cadherin complexes in human endothelial cells is regulated by thrombin. AB - Thrombin-mediated changes in endothelial cell adherens junctions modulate vascular permeability. We demonstrate that the nonreceptor protein-tyrosine phosphatase SHP2 co-precipitates with VE-cadherin complexes in confluent, quiescent human umbilical vein endothelial cells. Ligand-binding blots using a SHP2-glutathione S-transferase fusion peptide established that SHP2 associates selectively with beta-catenin in VE-cadherin complexes. Thrombin treatment of human umbilical vein endothelial cells promotes SHP2 tyrosine phosphorylation and dissociation from VE-cadherin complexes. The loss of SHP2 from the cadherin complexes correlates with a dramatic increase in the tyrosine phosphorylation of beta-catenin, gamma-catenin, and p120-catenin complexed with VE-cadherin. We propose that thrombin regulates the tyrosine phosphorylation of VE-cadherin associated beta-catenin, gamma-catenin, and p120-catenin by modulating the quantity of SHP2 associated with VE-cadherin complexes. Such changes in adherens junction complex composition likely underlie thrombin-elicited alterations in endothelial monolayer permeability. PMID- 10681593 TI - Constitutive death of platelets leading to scavenger receptor-mediated phagocytosis. A caspase-independent cell clearance program. AB - Apoptosis is a physiological program for the deletion of cells in which caspases govern events leading to safe clearance by phagocytes. However, a growing weight of evidence now suggests that not all forms of programmed cell death are caspase dependent. We now report a complete and constitutive but caspase-independent program for the specific phagocytic clearance of intact effete platelets, anucleated blood cells of critical importance in health and disease. Platelets aged in vitro not only exhibited increased expression of proapoptotic Bak and Bax but also evidenced constitutive diminution of function such as decreased aggregation to ADP, which was accelerated by culture in the absence of plasma. This abrogation of cell function in plasma-deprived platelets was associated with morphological and biochemical features similar to those of granulocyte apoptosis, that is, cytoplasmic condensation, plasma membrane changes including exposure of phosphatidylserine and the granule protein P-selectin, and recognition by phagocyte scavenger receptors. However, and in contrast with constitutive death of other inflammatory blood cells by apoptosis, these events were not affected by caspase inhibitors, nor was there evidence of caspase-3 activation either by hydrolysis of analog peptide substrates or Western blot analysis, serving to emphasize that neither programmed cell death nor clearance by phagocytes need involve caspases. PMID- 10681594 TI - Expression of distinct ERG proteins in rat, mouse, and human heart. Relation to functional I(Kr) channels. AB - One form of inherited long QT syndrome, LQT2, results from mutations in HERG1, the human ether-a-go-go-related gene, which encodes a voltage-gated K(+) channel alpha subunit. Heterologous expression of HERG1 gives rise to K(+) currents that are similar (but not identical) to the rapid component of delayed rectification, I(Kr), in cardiac myocytes. In addition, N-terminal splice variants of HERG1 and MERG1 (mouse ERG1) referred to as HERG1b and MERG1b have been cloned and suggested to play roles in the generation of functional I(Kr) channels. In the experiments here, antibodies generated against HERG1 were used to examine ERG1 protein expression in heart and in brain. In Western blots of extracts of QT-6 cells expressing HERG1, MERG1, or RERG1 (rat ERG1) probed with antibodies targeted against the C terminus of HERG1, a single 155-kDa protein is identified, whereas a 95-kDa band is evident in blots of extracts from cells expressing MERG1b or HERG1b. In immunoblots of fractionated rat (and mouse) brain and heart membrane proteins, however, two prominent high molecular mass proteins of 165 and 205 kDa were detected. Following treatment with glycopeptidase F, the 165- and 205-kDa proteins were replaced by two new bands at 175 and 130 kDa, suggesting that ERG1 is differentially glycosylated in rat/mouse brain and heart. In human heart, a single HERG1 protein with an apparent molecular mass of 145 kDa is evident. In rats, ERG1 protein (and I(Kr)) expression is higher in atria than ventricles, whereas in humans, HERG1 expression is higher in ventricular, than atrial, tissue. Taken together, these results suggest that the N-terminal alternatively spliced variants of ERG1 (i.e. ERG1b) are not expressed at the protein level in rat, mouse, or human heart and that these variants do not, therefore, play roles in the generation of functional cardiac I(Kr) channels. PMID- 10681595 TI - Isolation and characterization of cDNA clones for the e1beta and E2 subunits of the branched-chain alpha-ketoacid dehydrogenase complex in Arabidopsis. AB - Branched-chain alpha-ketoacid dehydrogenase (BCKDH) has been known in mammals to be a key enzyme of the catabolic pathway of branched-chain amino acids. We have isolated two cDNA clones encoding the E1beta and E2 subunits of BCKDH, respectively, from Arabidopsis thaliana. Proteins encoded in these cDNA sequences had putative mitochondrial targeting sequences and conserved domains reported for their mammalian counterparts. Northern blot and immunoblot analyses showed that transcripts from the respective genes and E2 protein markedly accumulated in leaves kept in the dark. We found that the activity of BCKDH in the leaf extracts also increased when plants were placed in the dark. Addition of sucrose to detached leaves inhibited the accumulation of transcripts, whereas application of a photosynthesis inhibitor strongly induced the expression of these genes even under light illumination. These observations indicate that the cellular sugar level is likely responsible for the dark-induced expression of these genes. The transcript levels of these genes were also high in senescing leaves, in which photosynthetic activity is low and free amino acids from degraded protein are likely to serve as an alternative energy source. PMID- 10681596 TI - Membrane targeting and cytoplasmic sequestration in the spatiotemporal localization of human protein kinase C alpha. AB - In order to map the molecular determinants that dictate the subcellular localization of human protein kinase C alpha (hPKCalpha), full-length and deletion mutants of hPKCalpha were tagged with the green fluorescent protein (GFP) and transiently expressed in GH3B6 cells. We found that upon thyrotropin releasing hormone (TRH) or phorbol 12-myristate 13-acetate stimulation, hPKCalpha GFP was localized exclusively in regions of cell-cell contacts. Surprisingly, PKCalpha failed to translocate in single cells despite the presence of TRH receptors, as attested by the TRH-induced rise in intracellular calcium concentration in these cells. TRH-stimulated translocation of hPKCalpha-GFP from the cytoplasm to cell-cell contacts was a biphasic process: a fast (measured in seconds) and transient phase, followed by a slower (approximately 1 hour) and long lasting phase. The latter and the translocation induced by phorbol 12 myristate 13-acetate absolutely required the N-terminal V1 region. In contrast to the full-length hPKCalpha, the N-terminal regulatory domain alone or associated with the V3 hinge region was spontaneously and uniformly localized at the plasma membrane of single and apposed cells. However, treatment with the calcium chelator BAPTA/AM induced a differential cytoplasmic/nuclear redistribution of the regulatory domain, depending on its association with V3, which suggests the existence of a mechanism controlling the cytoplasmic sequestration of inactive hPKCalpha and involving the V3 region. By using other deletion mutants, we were able to map the sequence required for this sequestration to the C2+V3 regions. This work points to the existence of a complex interplay between membrane targeting and cytoplasmic sequestration in the control of the spatiotemporal localization of hPKCalpha. PMID- 10681597 TI - Impaired kit- but not FcepsilonRI-initiated mast cell activation in the absence of phosphoinositide 3-kinase p85alpha gene products. AB - The class I(A) phosphoinositide 3-kinases (PI3Ks) consist of a 110-kDa catalytic domain and a regulatory subunit encoded by the p85alpha, p85beta, or p55gamma genes. We have determined the effects of disrupting the p85alpha gene on the responses of mast cells stimulated by the cross-linking of Kit and FcepsilonRI, receptors that reflect innate and adaptive responses, respectively. The absence of p85alpha gene products partially inhibited Kit ligand/stem cell factor-induced secretory granule exocytosis, proliferation, and phosphorylation of the serine/threonine kinase Akt. In contrast, p85alpha gene products were not required for FcepsilonRI-initiated exocytosis and phosphorylation of Akt. LY294002, which inhibits all classes of PI3Ks, strongly suppressed Kit- and FcepsilonRI-induced responses in p85alpha -/- mast cells, revealing the contribution of another PI3K family member(s). In contrast to B lymphocytes, mast cell proliferation was not dependent on Bruton's tyrosine kinase, a downstream effector of PI3K, revealing a distinct pathway of PI3K-dependent proliferation in mast cells. Our findings represent the first example of receptor-specific usage of different PI3K family members in a single cell type. In addition, because Kit- but not FcepsilonRI-initiated signaling is associated with mast cell proliferation, the results provide evidence that distinct biologic functions signaled by these two receptors may reflect differential usage of PI3Ks. PMID- 10681598 TI - The goodpasture autoantigen. Identification of multiple cryptic epitopes on the NC1 domain of the alpha3(IV) collagen chain. AB - Goodpasture (GP) disease is an autoimmune disorder in which autoantibodies against the alpha3(IV) chain of type IV collagen bind to the glomerular and alveolar basement membranes, causing progressive glomerulonephritis and pulmonary hemorrhage. Two major conformational epitope regions have been identified on the noncollagenous domain of type IV collagen (NC1 domain) of the alpha3(IV) chain as residues 17-31 (E(A)) and 127-141 (E(B)) (Netzer, K.-O. et al. (1999) J. Biol. Chem. 274, 11267-11274). To determine whether these regions are two distinct epitopes or form a single epitope, three GP sera were fractionated by affinity chromatography on immobilized NC1 chimeras containing the E(A) and/or the E(B) region. Four subpopulations of GP antibodies with distinct epitope specificity for the alpha3(IV)NC1 domain were thus separated and characterized. They were designated GP(A), GP(B), GP(AB), and GP(X), to reflect their reactivity with E(A) only, E(B) only, both regions, and neither, respectively. Hence, regions E(A) and E(B) encompass critical amino acids that constitute three distinct epitopes for GP(A), GP(B), and GP(AB) antibodies, respectively, whereas the epitope for GP(X) antibodies is located in a different unknown region. The GP(A) antibodies were consistently immunodominant, accounting for 60-65% of the total immunoreactivity to alpha3(IV)NC1; thus, they probably play a major role in pathogenesis. Regions E(A) and E(B) are held in close proximity because they jointly form the epitope for Mab3, a monoclonal antibody that competes for binding with GP autoantibodies. All GP epitopes are sequestered in the hexamer configuration of the NC1 domain found in tissues and are inaccessible for antibody binding unless dissociation of the hexamer occurs, suggesting a possible mechanism for etiology of GP disease. GP antibodies have the capacity to extract alpha3(IV)NC1 monomers, but not dimers, from native human glomerular basement membrane hexamers, a property that may be of fundamental importance for the pathogenesis of the disease. PMID- 10681599 TI - Metalloproteolytic release of endothelial cell protein C receptor. AB - Previous studies observed that there is about 100 ng/ml soluble endothelial cell protein C receptor (EPCR) in human plasma and that the levels increase in inflammatory diseases. In this study we examine the potential mechanisms involved in release of EPCR from cells. We find that EPCR is released from the surface of endothelium and transfected 293 cells by a metalloprotease in a constitutive fashion. The mass of soluble EPCR is 4 kDa less than intact EPCR. Release is blocked by either the hydroxamic acid based inhibitor, KD-IX-73-4 or by 1,10 phenanthroline, but not by matrix metalloprotease inhibitors. Release is stimulated by phorbol 12-myristate 13-acetate, thrombin, interleukin-1beta, and hydrogen peroxide. Stimulation with these agents reduces EPCR expression levels sufficiently to decrease the rate of protein C activation to a limited extent. The influence of phorbol 12-myristate 13-acetate on both EPCR release and inhibition of protein C activation are enhanced by microtubule disruption with nocodazole. EPCR release is augmented by transfection of EPCR expressing 293 cells with caveolin, suggesting that release is caveolae dependent. These studies indicate that metalloproteolytic release of EPCR is a highly regulated process that is sensitive to both coagulation factors and inflammatory mediators. PMID- 10681601 TI - Nitric oxide synthase expression in the opossum superior colliculus: a histochemical, immunohistochemical and biochemical study. AB - The expression of neuronal nitric oxide synthase (nNOS) in the superior colliculus (SC) of the opossum Didelphis marsupialis was studied by NADPH diaphorase (NADPH-d) histochemistry and nNOS immunohistochemistry. In addition, the activity of nNOS was quantified by measurement of [(3)H]-L-arginine conversion to [(3)H]-L-citrulline in tissue extracts from SC superficial layers in opossums and rats. Our results show that the number of NADPH-d stained cells was small and virtually identical in stratum opticum (SO) and stratum griseum superficiale (SGS) and their staining was very light, particularly in SGS. Neuropil staining was heavier in the stratum zonale (SZ) than in SGS or SO. The intermediate and deep layers contained heavily stained cells and moderate neuropil staining. Surprisingly, nNOS-immunoreactive cells were far more numerous than NADPH-d+ cells in every layer. The production of [(3)H]-L-citrulline from [(3)H]-L-arginine in tissue extracts enriched in superficial layers indicated that nNOS specific activity is as high in the opossum as in the rat. Our results suggest that the location of nNOS-expressing neurons in retino-receptive layers may be related to inter-specific differences in the processing of visual information. PMID- 10681602 TI - Relative medial and dorsal cortex volume in relation to foraging ecology in congeneric lizards. AB - The need to locate distributed resources such as mates, food, and nests is correlated with an enlarged hippocampus in many mammalian and avian species. This correlation is believed to be a consequence of selection for spatial ability. Little is known about how such ecological needs affect non-mammalian, non-avian species. In lizards, the putative hippocampal homologues are the dorsal cortex (DC) and medial cortex (MC). We examined the relationship between foraging ecology and the size of the DC and MC in congeneric male lizards. We predicted based on the mammalian and avian literature that Acanthodactylus boskianus, an active forager that captures clumped, immobile prey would have a larger MC and DC than A. scutellatus, a sit-and-wait predator, that captures mobile prey. Our previous behavioral studies showed that A. boskianus did not differ from A. scutellatus on a spatial task but that A. boskianus was significantly better at the reversal of a visual discrimination, another task that is hippocampally dependent in mammals. In the current study, we found that, relative to telencephalon volume, the MC and DC were larger in the active forager whereas a control region, the lateral, olfactory, cortex, was similar in size between species. The current anatomical results suggest that MC and DC size is related to active foraging in lizards and, along with our previous behavioral studies, show that it is possible for this relationship to occur in the absence of evidence for species differences in spatial memory. PMID- 10681603 TI - A possible pathway connecting the photosensitive pineal eye to the swimming central pattern generator in young Xenopus laevis tadpoles. AB - The pineal eye of young Xenopus laevis tadpoles mediates a swimming response to dimming. Our aim was to define pathways that allow pineal photoreceptors to influence the swimming central pattern generator (CPG) in the hindbrain and spinal cord. Retrograde filling with horseradish peroxidase (HRP) and carboxyfluorescein showed that: (1) pineal ganglion cells do not project to the hindbrain, and (2) diencephalic/mesencephalic descending (D/MD) neurons, which could be contacted by pineal ganglion cell axons, do project to the hindbrain. Lesion experiments demonstrated that ganglion cell axons form ipsilateral and contralateral connections, either of which is sufficient to mediate a swimming response. Latency measurements suggest that the contralateral pathway is stronger than the ipsilateral one. Multiple unit recordings from the midbrain in the region of the D/MD neurons showed short latency activity in response to dimming or a brief current pulse to pineal axons. This activity could last for many seconds after the stimulus. Pharmacological experiments showed that it depended on synaptic excitation and suggested that the ganglion cell transmitter is glutamate. If pineal ganglion cells excite midbrain D/MD neurons on both sides of the brain, the D/MD neuron projections to the hindbrain could excite the swimming CPG and initiate swimming. PMID- 10681604 TI - Responses of single neurons in the toad's caudal ventral striatum to moving visual stimuli and test of their efferent projection by extracellular antidromic stimulation/recording techniques. AB - Previous work in anuran amphibians has shown that activity in the caudal ventral striatum correlates with visuomotor activity: orienting responses toward prey fail to occur after striatal lesions. Thus it has been suggested that the striatum influences visually guided behavior. Therefore, the present study investigates visual response properties from neurons recorded in the striatum. Extracellular recordings of 104 single neurons of the cane toad's (Bufo marinus) caudal ventral striatum (STR) reveal five different response properties: resting discharge activity uninfluenced by the visual test stimuli (group STR1, 24.0%); resting discharge activity increased by any moving visual object (STR2, 31.7%); preference to moving compact objects (STR3, 15.4%); preference to certain configurational moving objects (STR4a and b, 13.5%), and resting activity reduced by visual stimuli (STR5, 15.4%). The receptive fields of these neurons encompassed the contralateral (46%) or the entire field of vision (54%). Of the neurons recorded in the striatum, 34% responded to electrical stimuli applied in the rostral diencephalon to the ipsilateral lateral forebrain bundle (LFB) which connects the striatum with the optic tectum (e.g. either directly or via pretectum or tegmentum). Various electrically driven STR neurons (40%) have axons that project caudally through the LFB, which was suggested by their antidromic activation in response to electrical stimuli applied to the LFB in the rostral diencephalon. In the present study, the main striatal output is mediated by 'motion detectors' (STR2) and 'compact object perceivers' (STR3). It is suggested that the caudal ventral striatum is involved in visual attentional processes that allow the translation of perception into action. PMID- 10681605 TI - Personal UV dosimetry by Bacillus subtilis spore films. AB - BACKGROUND: Ultraviolet radiation (UVR) is known to be the most important risk factor for melanoma and non-melanoma skin cancers. Until today it has been impossible to measure reliably UVR in the frame of epidemiological studies. The recent development of a spore film containing spores of Bacillus subtilis resulted in a new method of UV measurement by personal dosimetry. METHODS: The practical application of dosimeters was tested in 18 study persons under different circumstances of UV exposure and in 4 different geographical regions. RESULTS: Eleven children carried dosimeters on their shoulders for 1 day, playing in- and outdoors on a sunny day in summertime. Their whole-day values ranged from 0.1 to 1.5 minimal erythema doses (MED) per day with a mean of 0.71 MED (+/ 0.44). Four lifeguards in a public swimming-pool carried dosimeters on their shoulders for 11 days and received UV exposures ranging from 3.6 to 9.5 MED (mean 5.9 +/- 1.88). Three mountain guides with dosimeters attached to the lateral head in different mountain regions at 23 mountaineering activities received daily exposures of 4.44-17.07 MED (mean 11.9 +/- 3.8). CONCLUSION: B. subtilis spore film dosimeters can be applied to different study persons including children and mountain guides under different climatic conditions. A broad range of UV exposures can reliably be measured with this method. PMID- 10681606 TI - Clinical, histological and immunopathological features of 58 patients with subacute cutaneous lupus erythematosus. A review by the Italian group of immunodermatology. AB - BACKGROUND: Subacute cutaneous lupus erythematosus (SCLE) is a distinct subset of cutaneous lupus erythematosus clinically characterized by psoriasiform and/or annular lesions and by a mild or absent systemic involvement. OBJECTIVE: The Italian Group of Immunodermatology of the Italian Society of Dermatology and Venereology reviewed the cases of SCLE seen in 10 years (1987-1996). PATIENTS: Forty-six women and 12 men have been retrospectively studied, 42% had annular lesions, 39% psoriasiform ones and 16% both. RESULTS: Lesions were mainly localized on the neck and face and relapsed in spring and autumn. Seventeen patients had 4 or more American College of Rheumatology criteria and could be classified as having systemic lupus erythematosus. The most frequent histopathological alterations were epidermal atrophy, hydropic degeneration of the basal layer and perivascular lymphocytic infiltrate. Deposits of immunoglobulins and C3 at the dermo-epidermal junction on the clinically involved skin were present in 86% of the patients. Dust-like particles in the epidermis were only found in 3% of cases. Anti-Ro/SSA antibodies were found in 71% of the cases and anti-dsDNA only in 5% of cases. CONCLUSIONS: SCLE is a particular subset of cutaneous lupus erythematosus with peculiar clinical and immunopathological features. PMID- 10681607 TI - A cancer-registry-assisted evaluation of the accuracy of digital epiluminescence microscopy associated with clinical examination of pigmented skin lesions. AB - BACKGROUND: The accuracy of digital epiluminescence microscopy (D-ELM) as an adjunct to clinical examination for the diagnosis of pigmented skin lesions (PSLs) has seldom been evaluated. OBJECTIVE: To compare the accuracy of the combined clinical/D-ELM (C/D-ELM) examination with that of the clinical examination alone. METHODS: A total of 3,372 PSLs from 1,556 consecutive patients referred to a skin cancer clinic underwent clinical examination and a combined C/D-ELM examination. The reference diagnosis was established using the histology report of known surgical excisions plus a cancer-registry-based follow-up (duration 18 months) of benign C/D-ELM diagnoses. The two diagnostic approaches were compared for sensitivity, predictive value and false-positive rate. RESULTS: The series included 55 melanomas and 43 basal cell carcinomas. About 50% of malignant misdiagnosed cases were identified solely through the cancer registry. The C/D-ELM diagnosis showed a greater sensitivity for melanoma <0.76 mm thick (83 vs. 46% for clinical examination alone; ratio, 1.82) and basal cell carcinoma (79 vs. 49%; ratio, 1. 62), a greater predictive value for melanoma (81 vs. 53%; ratio, 1. 53) and a reduced total false-positive rate (0.3 vs. 0.9%; ratio, 0. 31). CONCLUSION: D-ELM showed a potential to improve the clinical diagnosis of PSL. PMID- 10681608 TI - Calcipotriol cream combined with twice weekly broad-band UVB phototherapy: a safe, effective and UVB-sparing antipsoriatric combination treatment. The Canadian Calcipotriol and UVB Study Group. AB - BACKGROUND: Calcipotriol has been combined with a number of systemic antipsoriatric treatments, improving efficacy or reducing the systemic treatment required. Although studies on calcipotriol and UVB have also been performed, there are no data on the UVB-saving effect of calcipotriol combined with broad band UVB to reduce overall UVB exposure, while maintaining efficacy. OBJECTIVES: To assess the efficacy and safety of calcipotriol cream (50 microg/g) combined with twice weekly broad-band UVB and to determine if this treatment would require fewer UVB treatments and lower cumulative UVB irradiance when compared to a standard 3 times weekly broad-band UVB regime in patients with extensive psoriasis. METHODS: This multicentre, prospective, randomised, parallel-group, vehicle-controlled, single-blind (investigator) study consisted of a 1-week wash out phase, 12-week treatment phase and 12-week follow-up phase. Broad-band UVB equipment was standardised and calibrated prior to the study. The UVB starting dose was based on the patient's minimal erythema dose. Assessments included PASI, extent, severity and investigator and patient's overall assessments of the psoriasis. RESULTS: Fewer exposures (12 vs. 19) and less cumulative UVB irradiance (1,570 vs. 5,430 mJ/cm(2)) were required by the calcipotriol + twice weekly UVB group to achieve 80% reduction in PASI (p < 0.001). Similarly, fewer exposures (22 vs. 25) and less cumulative UVB irradiance (4,147 vs. 9,670 mJ/cm(2)) were required by this group to achieve total clearance (p < 0.001). There was no difference in the PASI, patient's and investigator's overall assessments and number of adverse events recorded by either group for both the treatment and follow-up phases. CONCLUSION: Calcipotriol cream + twice weekly broad-band UVB phototherapy is an effective and safe antipsoriatric treatment, resulting in fewer UVB exposures, lower cumulative irradiance and a saving of time. PMID- 10681609 TI - Efficacy of calcipotriol ointment applied under hydrocolloid occlusion in psoriasis. AB - BACKGROUND: Hydrocolloid (HCD) dressings enhance the efficacy of topical corticosteroids. OBJECTIVE: We wanted to evaluate the effect of calcipotriol ointment under an HCD dressing in the treatment of psoriatic plaques. METHODS: In 9 psoriatic patients, we cleared one plaque using this approach and took biopsies at start, clearance and relapse. Clinical and immunohistochemical validation was assessed. RESULTS: After an average treatment of 3.6 weeks, each lesion had cleared (apart from some residual erythema). The average remission period was 8 weeks. During this treatment, the number of cycling epidermal cells (Ki-67 positive nuclei) and the expression of keratin 14 and keratin 16 had decreased substantially. In biopsies taken from the skin immediately adjacent to the relapsing lesion, these markers remained reduced which indicated the prolonged effect of calcipotriol on epidermal differentiation. CONCLUSION: It is speculated that combination therapy of calcipotriol with treatments with a different mode of action such as photo(chemo)therapy, corticosteroids and cyclosporine might be worthwhile. PMID- 10681610 TI - Treatment and pain relief of ulcerative hemangiomas with a polyurethane film. AB - BACKGROUND: Hemangiomas are the most common tumors occurring in young children. The most common complication in the growing phase of hemangioma is ulceration. AIM AND METHOD: We report healing, pain relief and evolutive effects of a polyurethane film in 8 cases with ulcerative hemangiomas. RESULTS: In all 8 infants, prompt pain relief and healing within 1-2 months were observed. An increased regression was also noted within 2-4 months, when the hemangiomas were in the normal proliferative phase. CONCLUSION: As far as the authors know, there is no explanation for the effectiveness of polyurethane film. Explanations could be the occlusive effects of the film inhibiting proliferation or the decrease in blood flow. As primary initial therapeutic approach in ulcerative hemangiomas, we advocate the application of a polyurethane film. This therapy is painless and suitable for children. PMID- 10681611 TI - Clinical efficacy of narrow-band UVB (311 nm) combined with dithranol in psoriasis. An open pilot study. AB - BACKGROUND: For UVB, the most effective wavelength in clearing psoriatic lesions was found to be of 313 nm. The efficacy of whole body exposure to narrow-band UVB (311 nm) combined with dithranol in psoriasis has not been evaluated to date. OBJECTIVE: Evaluation of the clinical efficacy of phototherapy with narrow-band UVB (311 nm) and dithranol for psoriasis by means of whole body exposures and analysis of the mean cumulative irradiation dose. METHODS: In this open pilot study, 13 patients were treated for 4-5 weeks. Evaluation of the therapeutic efficacy was performed by comparing the Psoriasis Area and Severity Index (PASI) scores at baseline and after 4 weeks of treatment. The cumulative irradiation dose was also calculated. RESULTS: Evaluation of the PASI scores showed a significant overall reduction of psoriatic lesions after 4 weeks of treatment. The cumulative irradiation dose was similar or lower to those found for phototherapy with narrow-band UVB alone. DISCUSSION: In patients with widespread psoriasis, treatment with narrow-band UVB (311 nm) combined with dithranol is safe and effective, allowing reduction of the cumulative irradiation dose. PMID- 10681612 TI - Paronychia with pyogenic granuloma in a child treated with indinavir: the retinoid-mediated side effect theory revisited. AB - BACKGROUND: The introduction of HIV-1 protease inhibitors into the treatment of patients infected with HIV-1 has had a major influence on clinical practice. However, the use of protease inhibitors is frequently associated with the development of resistance and several side effects and interactions with other drugs have been reported. OBSERVATIONS: We present the first pediatric patient with paronychia with pyogenic granuloma associated with the administration of the protease inhibitor indinavir. Clinical findings are discussed in view of a possible interference of indinavir with endogenous retinoid metabolism. CONCLUSION: Considerable evidence advocates the mediation of indinavir side effects by impaired oxidative metabolism of retinoic acid through the inhibition of cytochromes P450 3A by indinavir rather than by impaired formation of 9-cis retinoic acid. PMID- 10681613 TI - A prospective study of acute-onset steroid acne associated with administration of intravenous corticosteroids. AB - Steroid acne (SA) may occur after the administration of topical or systemic corticosteroids. Because of several consultations of spinal injury patients with a very abrupt onset of a uniform papular eruption (i.e. days) initially misdiagnosed as a drug reaction or sepsis, we followed hospitalized patients who received intravenous corticosteroids (IVC) for the development of acute-onset SA in order to determine its incidence. Fifty-one consecutive subjects receiving IVC were followed for the duration of their hospital stay and examined for the development of acneiform lesions. Acute-onset SA occurred in 1 subject (2%). Acute spinal cord injury may represent a high-risk clinical setting for acute onset SA. PMID- 10681614 TI - Leg ulcers in patients with myeloproliferative disorders: disease- or treatment related? AB - Leg ulcers are a relatively frequent problem in patients with myeloproliferative disorders under treatment with hydroxyurea (HU). The pathogenesis is currently unknown and may be multifactorial. Concomitant arterial or venous disease may play a contributing role in the development of these wounds. Vasculitis, cryoglobulinemia and pyoderma gangrenosum should be considered if typical clinical signs are present. We report on 3 patients with myeloproliferative disorders who developed HU-induced leg ulcers and review the literature. HU induced leg ulcers share clinical features which can help to differentiate them from leg ulcers of other etiologies: occurrence under long-term treatment with HU at a dose of at least 1 g/day, localization in the malleolar region and spontaneous healing when HU is discontinued. We conclude that differentiation between disease-related and treatment-induced leg ulcers can be difficult and may not always be possible. In HU-induced leg ulcers, cessation of the drug typically leads to wound healing. PMID- 10681615 TI - Kaposi's sarcoma after lung transplantation in a Sephardic Jewish woman. AB - One of the four types of Kaposi's sarcoma (KS), KS after organ transplantation under immunosuppression, is a well-known entity and has been abundantly described in renal, heart and liver recipients. We report the second case of cutaneous KS after lung transplantation, under regular immunosuppression, in a Sephardic Jewish woman. This case, when added to the other 10 cases of posttransplantation KS reported from Israel, all being Sephardic Jews, indicates that in Israel, Sephardic Jews are at higher risk than Ashkenazi Jews to develop posttransplantation KS. This observation should be added to the well-known increased risk of Ashkenazi Jews to develop classic KS. Moreover, in Israel Ashkenazi Jews develop classic KS at higher rates than Sephardic Jews. This apparent discrepancy in the ethnic distribution between Sephardic and Ashkenazi Jews in classic versus posttransplantation KS may shed light on the pathogenesis of KS in general. PMID- 10681616 TI - Interferon-alpha-associated development of bullous lesions in mycosis fungoides. AB - We report a 67-year-old woman with mycosis fungoides (MF) who was receiving subcutaneous injections of recombinant interferon alpha-2b (IFN-alpha). After 2 months of IFN-alpha treatment, bullous lesions appeared on her trunk and extremities. A skin biopsy specimen from the trunk revealed histopathologic features of bullous MF. Bullous lesions with specific infiltrates of MF are very rare; to the best of our knowledge, this is the first case showing specific bullous lesions of MF developed under IFN-alpha treatment. PMID- 10681617 TI - Generalized granuloma annulare associated with granulomatous mycosis fungoides. AB - We describe a 68-year-old man with plaque stage mycosis fungoides (MF) for 8 years. He developed tumorous lesions of granulomatous MF (GrMF) and generalized granuloma annulare (GA) after a previously indolent clinical course. Since then, the clinical course was aggressive with involvement of the bone marrow and lymph nodes, and leukemic change occurred. Systemic chemotherapy was given, but the patient died 9 months later due to neutropenic fever and septic shock. GA in malignant lymphoma has been reported most frequently in association with Hodgkin's disease. To the best of our knowledge, GA associated with GrMF has never been reported in the English language literature. The prognostic significance of the association of granulomatous inflammation and malignancy is reviewed. PMID- 10681618 TI - Pigmentation and pits at uncommon sites in a case with reticulate acropigmentation of Kitamura. AB - Reticulate acropigmentation of Kitamura is now reported from all over the world. Additional features are being readily recognized. Our cases had pigmentation and pits on the dorsa of the distal phalanges of the fingers and toes - the classical features - as well as widely distributed pits on the palms, palmar aspect and sides of the fingers. The involvement in our cases was more pronounced than in the previously reported ones. PMID- 10681619 TI - Tropical-wood-induced bullous erythema multiforme. AB - We report a case of bullous erythema multiforme caused by an exotic wood, pao ferro (Machaerium scleroxylon). A 25-year-old female, a luthier (guitar maker) who often handles a variety of woods, developed bullous erythema multiforme. A patch test confirmed a positive reaction to one of the exotic woods, pao ferro. A subsequent accidental short contact with pao ferro 5 months following the first incidence induced a similar exudative erythema. Exotic woods such as pao ferro should be added to the list of contact allergens that can induce bullous erythema multiforme. PMID- 10681620 TI - Primary atrophic profound linear scleroderma. Report of three cases. AB - We present 3 unusual cases of deep linear, primary atrophic scleroderma, not preceded by inflammatory reaction and sclerosis, involving the subcutis and deeper tissues. These cases differ in the course and prognosis from typical profound scleroderma since they do not lead to disfiguration and crippling deformities. In contrast to the atrophies left after regression of morphea or linear scleroderma, they do not involve the dermis, which does not show discoloration or changes in texture. Infiltrates in the endomysium, involvement of deeper tissues and the progressive character of the disease argue for atypical primary atrophic profound scleroderma. The coexistence in one case of primary facial hemiatrophy appears to indicate also its relationship with primary linear atrophies of the limbs. PMID- 10681621 TI - Postirradiation morphea of the breast presentation of two cases and review of the literature. AB - The advent of radiation therapy as a common modality in the treatment and palliation of breast cancer has led to the observation of morphea developing months to years after supervoltage radiation therapy, in and around the site of treatment. We report 2 new cases of morphea at the site of previous supervoltage radiation therapy for breast cancer. The time period between irradiation and onset of morphea in our 2 patients were an atypically long 6.5 years and 32 years, the latter being the longest reported such interval. With conservative treatment, the inflammatory component of the lesions resolved over an approximately 1-year period, leaving residual sclerosis. These patients are compared to those previously reported in the medical literature so as to summarize the range of clinical presentation and course. Recognition of postirradiation morphea is important in distinguishing it from infectious cellulitis, recurrent carcinoma, metastatic carcinoma or development of a second primary carcinoma. PMID- 10681622 TI - Recurrent intravascular papillary endothelial hyperplasia of the toes. AB - We report on the rare case of a 50-year-old male with intravascular papillary endothelial hyperplasia (IPEP) in the left toes. The patient noticed a small tender mass in the left toes and underwent surgery in November 1997. The tumor recurred twice after surgery. Histopathological examination revealed a pure form of IPEP without underlying benign vascular lesions. PMID- 10681623 TI - Three cases of type 2 segmental manifestation of multiple glomus tumors: association with linear multiple trichilemmal cysts in a patient. AB - We report 3 cases of congenital multiple glomus tumors seen during the last 5 years. One of them showed autosomal dominant inheritance with male-to-male transmission. The remaining patients had no family history of similar lesions. The clinical and histopathological aspects of our patients support the recently described type 2 segmental manifestation of multiple glomus tumors. One of the cases showed associated multiple and giant trichilemmal cysts with a linear distribution in the scalp. PMID- 10681624 TI - Humoral response in a patient with cutaneous nocardiosis. AB - The clinical appearance of infection due to Nocardia spp. varies widely. The low sensitivity of direct microscopy and the slow growth of the organism challenge the laboratory diagnosis. We present the case of a skin abscess in an immunocompetent man caused by Nocardia brasiliensis. Diagnosis was made by cultivation and 16S rRNA sequencing. Using indirect immunofluorescence and Western blot, a strong antibody response to the N. brasiliensis isolate could be demonstrated. Serological tests might therefore be useful for the diagnosis and management of nocardial infections. PMID- 10681625 TI - UV transmission of summer clothing in Switzerland and Germany. PMID- 10681626 TI - Macular amyloidosis due to friction by a horsehair glove. PMID- 10681627 TI - Serum IgA autoantibodies in patients with epidermolysis bullosa acquisita: a high frequency of detection. PMID- 10681628 TI - Total anonychia congenita in a woman with normal intelligence: report of a further case. PMID- 10681629 TI - Pigmentary demarcation lines over the face. PMID- 10681630 TI - Compression ulcer in a caisson worker. PMID- 10681631 TI - Lack of detection of leukaemia inhibitory factor in the sera of psoriatic patients. PMID- 10681632 TI - Papular-purpuric 'Gloves-and-Socks' syndrome related to rubella virus infection. PMID- 10681633 TI - Maternal hypothyroidism and child development. A review. PMID- 10681634 TI - Insulin-like growth factor binding protein (IGFBP)-3-bound IGF-I and IGFBP-3 bound IGF-II in growth hormone deficiency. AB - In blood, circulating IGFs are bound to six high-affinity IGFBPs, which modulate IGF delivery to target cells. Serum IGFs and IGFBP-3, the main carrier of IGFs, are upregulated by GH. The functional role of serum IGFBP-3-bound IGFs is not well understood, but they constitute the main reservoir of IGFs in the circulation. We have used an equation derived from the law of mass action to estimate serum IGFBP-3-bound IGF-I and IGFBP-3-bound IGF-II, as well as serum free IGF-I and free IGF-II, in 129 control children and adolescents (48 girls and 81 boys) and in 13 patients with GHD. Levels of serum total IGF-I, total IGF-II, IGFBP-1, IGFBP-2 and IGFBP-3 were determined experimentally, while those of IGFBP 4, IGFBP-5 and IGFPB-6, as well as the 12 affinity constants of association of the two IGFs with the six IGFBPs, were taken from published values. A correction for in vivo proteolysis of serum IGFBP-3 was also considered. In controls, serum total IGF-I, total IGF-II, IGFBP-3, IGFBP-3-bound IGF-I, IGFBP-3-bound IGF-II and free IGF-I increased linearly with age, from less than 1 to 15 years, in the two sexes. The concentrations of serum free IGF-I and free IGF-II were approximately two orders of magnitude below published values, as well as below the affinity constant of association of IGF-I with the type-1 IGF receptor. Therefore, it is unlikely that these levels can interact with the receptor. In the 13 patients with GHD, mean (+/- SD) SDS of serum IGFBP-3-bound IGF-I was -2.89 +/- 0.97. It was significantly lower than serum total IGF-I, free IGF-I or IGFBP-3 SDSs (-2.35 +/- 0.83, -1.12 +/- 0.78 and -2.55 +/- 1.07, respectively, p = 0.0001). The mean SDS of serum total IGF-II, IGFBP-3-bound IGF-II and free IGF-II were -1.25 +/- 0.68, -2.03 +/- 0.87 and 0.59 +/- 1.10, respectively, in GHD. In control subjects, 89.8 +/- 4.47% of serum total IGF-I and 77.3 +/- 9.4% of serum total IGF-II were bound to serum IGFBP-3. In patients with GHD, the mean serum IGFBP-3 bound IGF-I and IGFBP-3-bound IGF-II were 8.63 +/- 8. 53 and 19.1 +/- 14.7% below the respective means of control subjects (p < 0.02). In conclusion, in GHD there was a relative change in the distribution of serum IGFs among IGFBPs, due to the combined effects of the decrease in both total IGF-I and IGFBP-3. As a result, serum IGFBP-3-bound IGF-I and IGFBP-3 bound IGF-II, the main reservoirs of serum IGFs, were severely affected. This suggests that the decrease in serum IGFPB-3 bound IGF-I and IGFBP-3-bound IGF-II might have a negative effect for growth promotion and other biological effects of IGF-I and IGF-II. Finally, the estimation of serum IGFBP-3-bound IGF-I, or the percentage of total IGF-I and IGF II bound to IGFBP-3, might be useful markers in the diagnosis of GHD. PMID- 10681635 TI - Growth hormone treatment downregulates serum leptin levels in children independent of changes in body mass index. AB - The changes in serum leptin levels during growth hormone (GH) treatment were studied in 27 children, 17 with GH deficiency (GHD), 10 with idiopathic short stature (ISS), and 9 with Prader-Willi syndrome (PWS). Within 1 month of GH treatment, serum leptin levels decreased by 40% in the GHD children (p < 0.01). There was no significant change in serum leptin level in the children with ISS. In children with PWS, the mean serum leptin level decreased by almost 60% after 3 months of treatment (p < 0.001). Thereafter, no further decline was observed in any of the 3 groups. Changes in body composition became evident first after the 3 months of treatment. In the GHD children, the BMI was unchanged while the mean body fat percentage was 2.7% lower after 1 year of GH treatment (p < 0.05). In the ISS children, neither BMI nor body fat percentage were significantly changed during treatment. The PWS children exhibited a significant decrease in BMI after 6 months of GH treatment without any further change during the remaining period of treatment. In this group, the mean body fat percentage decreased from 42 +/- 2.4 to 28 +/- 2.2% after treatment (p < 0.001). The finding that the fall in leptin occurs before changes in body composition become detectable suggests a direct effect of GH on leptin production, metabolism, or clearance. PMID- 10681636 TI - Diagnosis of ACTH deficiency. Comparison of overnight metyrapone test to either low-dose or high-dose ACTH test. AB - Test sensitivity and accuracy of 250 microg/m(2) ACTH test, 1 microg/m(2) ACTH test, and overnight metyrapone test were evaluated in 158 children at risk for ACTH deficiency. Of 38 given high-dose ACTH, 20 had normal responses to metyrapone and to high-dose ACTH. 14 had low response to metyrapone; of these only 2 had low cortisol response (<550 nmol/l) to high-dose ACTH. Of 120 given low-dose ACTH, 64 had normal responses to metyrapone and to low-dose ACTH. All 24 with low metyrapone response had low or borderline response to low-dose ACTH. The remaining children had an inconclusive metyrapone response. In conclusion, high dose ACTH misses most diagnoses of ACTH deficiency (21% sensitivity, 100% specificity, 63% accuracy). In contrast, the low dose ACTH test accurately diagnoses 90% of patients with ACTH deficiency (100% sensitivity, 68% specificity). The low-dose ACTH test can serve as an accurate and practical screening test for adequacy of ACTH reserve. PMID- 10681637 TI - Final height, armspan, subischial leg length and body proportions in juvenile chronic arthritis. A long-term follow-up study. AB - OBJECTIVE: Assessment of growth disturbances in adults with a history of juvenile chronic arthritis (JCA). MATERIAL AND METHODS: Sixty-five subjects, 52 premenopausal females and 13 males with a mean age (range) of 32.2 years (22.3 49.4) participated. Mean age at disease onset was 5.7 years (0.8-15.8) and mean disease duration was 12.4 years (0.4-32). The follow-up time ranged from 18.7 to 46.9 years with a mean of 26.4 years. For each participant standard deviation scores (z-scores) for final height, delta-height (the difference between observed and expected height), armspan, subischial leg length and sitting height ratio, were calculated. RESULTS: The study group as a whole did not exhibit linear growth impairment. The categorical distribution of heights differed significantly from a expected distribution in a healthy population (p < 0.001). A height z score < -2 SD was present in 10.7% of the study group, of whom all had polyarticular course of JCA. Polyarticular and systemic course of JCA (versus pauciarticular) (p = 0.022), systemic steroid treatment (p = 0.006) and Steinbrocker functional class II-IV (vs. I) in 1979 (p = 0.043) were variables associated with reduced delta-height. In linear regression analyses, disease severity defining variables were statistically significant predictors of reduced final height and armspan. 27% of the study subjects had significantly reduced arm span (p < 0.001). Subischial leg length and body proportions (sitting height ratio) were normal. CONCLUSION: Our findings suggest that functionally impaired polyarticular and systemic JCA patients treated with systemic steroids may be at an increased risk of developing reduced final height and armspan. Disease control achieved by an aggressive therapeutic approach, if possible with a minimal use of systemic steroids, may reduce growth impairment in JCA. PMID- 10681638 TI - Sera from children with type 1 diabetes mellitus react against a new group of antigens composed of lysophospholipids. AB - Several autoantibodies related to Type 1 diabetes mellitus and their corresponding autoantigens have been previously identified. While peptide antigens are more widely recognized, lipid antigens like sulfatides and gangliosides are also known epitopes for the diabetic humoral immune response. Islet cell antibodies (ICA) in Type 1 diabetes are heterogeneous immunoglobulins directed against selected antigens in the islets of Langerhans. Moreover, ICA may be the best predictive marker of disease in family members of patients with Type 1 diabetes. The aims of this study were: (1) to purify lipids from porcine pancreas that contain ICA epitopes; (2) to characterize these lipid antigens, and (3) to use the purified lipids in an assay to detect antibodies in patients with Type 1 diabetes. A unique family of 4 lysophospholipids, 1 fully characterized as lysophosphatidylmyoinositol, partially inhibited ICA staining, and therefore, were considered to be candidate antigens for an ICA immunoassay. Using a dot blot immunoassay, we detected antibodies directed against these phospholipids in 28 out of 46 (61%) diabetic sera, while detecting only 1 false positive out of 28 nondiabetic sera (3.6%; p < 0.0001 comparing diabetic vs. nondiabetic serum). Therefore, lysophospholipid immunoassay positivity is present in sera of Type 1 diabetic patients. Furthermore, we detected 15 out of 23 ICA-negative diabetic sera (65.2%), showing that our phospholipid immunoassay does not correlate with ICA positivity. PMID- 10681639 TI - Growth-suppressive effect of intra-articular glucocorticoids detected by knemometry. AB - After one intra-articular injection of 20 mg triamcinolone hexacetonide in the knee, the length of the contralateral lower leg was found to be reduced in 2 boys with juvenile rheumatoid arthritis. PMID- 10681640 TI - Peritoneal carcinomatosis following laparoscopic resection of an adrenocortical tumor causing primary hyperaldosteronism. AB - A clinical syndrome combining hypertension and hypokalemic alkalosis led to the diagnosis of primary hyperaldosteronism, caused by a right-sided, 2 cm large, apparently benign aldosterone-producing adenoma. The adrenal tumor was completely resected by laparoscopic adrenalectomy. Six months after surgery, the patient exhibited a severe relapse of hyperaldosteronism. Extensive peritoneal metastases of a mixed aldosterone- and cortisol-secreting adrenocortical carcinoma were found at abdominal laparotomy. In the light of this case report, we discuss the possibility that laparoscopic resection of adrenocortical tumors might contribute to their subsequent peritoneal dissemination. PMID- 10681641 TI - Acute watery diarrhea as the initial presenting feature of a pheochromocytoma in an 84-year-old female patient. AB - We report the case of an 84-year-old woman who was initially admitted to the emergency room of our institution for frank dehydration caused by acute and severe secretory diarrheas along with acidosis and hypokalemia. After extensive gastrointestinal investigations, the etiology of the diarrhea remained unclear. Because clinical symptoms and ionogram parameters worsened, despite intravenous fluids and electrolyte replacement, an abdominal CT scan was performed and unexpectedly revealed a 4.5-cm mass in the right adrenal gland. Several separate 24-hour urine catecholamines were shown to be highly elevated. The diagnosis of pheochromocytoma was confirmed by MIBG scintigraphy and MRI. Before the admission, the patient never experienced symptoms suggestive of pheochromocytoma, except dry mouth and fear of impending death on several occasions. After 2 weeks, the diarrhea stopped abruptly and spontaneously without specific medication but after adequate rehydration. The patient subsequently underwent surgical removal of the adrenal medullary mass. Postoperatively, urinary catecholamines returned to normal values. Immunohistochemical study of the tumor confirmed the diagnosis of pheochromocytoma and revealed the presence of VIP-positive cells organized as islets in scattered areas of the tissue. This case illustrates the protean mode of presentation of pheochromocytoma, as well as the ability of medullary neural crest-derived cells to produce various neuropeptides potentially responsible for a large variety of symptoms. PMID- 10681642 TI - Mechanisms causing muscle proteolysis in uremia: the influence of insulin and cytokines. AB - Decreased muscle mass in patients with chronic renal failure (CRF) can be caused by mechanisms that activate the ubiquitin-proteasome proteolytic system. This system accelerates the degradation of muscle protein. Concurrent with muscle protein breakdown, there is an increase in transcription of genes encoding components of this pathway, including ubiquitin and subunits of the proteasome. Potential activating signals include metabolic acidosis which stimulates proteolysis in CRF patients and in muscle of rats with CRF by a mechanism involving glucocorticoids. In CRF patients, there is insulin resistance and high circulating levels of tumor necrosis factor and other cytokines. As the ubiquitin proteasome proteolytic system is activated in acute diabetes and in catabolic conditions associated with high levels of circulating cytokines, these factors could also activate this pathway. Consequently, we examined whether the transcription factor activated by certain cytokines, NF-kappaB, is involved in the transcriptional regulation of subunits of the 26S proteasome complex. The results suggest that cytokines may be involved in the regulation of muscle protein degradation in uremia. PMID- 10681643 TI - The balance between glucocorticoids and insulin regulates muscle proteolysis via the ubiquitin-proteasome pathway. AB - In uremia, accelerated muscle protein degradation results from activation of the ATP-ubiquitin proteasome proteolytic pathway. Like uremia, other conditions (e.g., acidosis and diabetes) activate this pathway in rat muscles and are associated with excess glucocorticoids (GC) and impaired insulin action. To define the stimuli responsible for muscle wasting in IDDM, the roles of glucocorticoids, insulinopenia and acidosis in streptozotocin (STZ) - induced diabetes were studied. Proteolysis in isolated epitrochlearis muscles from acutely (3d) diabetic rats was 52% higher than pair-fed, sham-injected rats; this increase was eliminated by an inhibitor of the proteasome or by blocking ATP synthesis. In muscles of STZ-diabetic rats, the levels of ubiquitin-conjugated proteins and mRNAs encoding ubiquitin, the ubiquitin-carrier protein, E2(14k) and the C3, C5 and C9 proteasome subunits were increased. Transcription of ubiquitin and C3 proteasome subunit genes in muscle was also increased by IDDM. Oral NaHCO(3) eliminated acidemia but did not prevent accelerated muscle proteolysis. Corticosterone excretion was higher in IDDM rats and adrenalectomy (ADX) prevented these catabolic responses; physiologic doses of glucorcoticoids restored the excessive protein catabolism in ADX-STZ rats. Giving IDDM rats replacement insulin also normalized protein degradation in muscles. In conclusion, reduced insulin together with physiologic levels of glucocorticoids activate the ubiquitin-proteasome pathway by a mechanism that includes enhancing ubiquitin conjugation and proteolysis by the proteasome. The balance between these stimuli could regulate muscle proteolysis in uremia. PMID- 10681644 TI - Glucocorticoids and acidification independently increase transcription of branched-chain ketoacid dehydrogenase subunit genes. AB - Metabolic acidosis and glucocorticoids act in concert to stimulate branched-chain amino acid (BCAA) oxidation in adrenalectomized rats. In muscles of normal rats, metabolic acidosis increases the maximal activity of the rate-limiting enzyme, branched-chain alpha-ketoacid dehydrogenase (BCKAD) and a genetic response to catabolic conditions like uremia is implicated by concurrently higher levels of BCKAD subunit mRNA. To determine if acidification or glucocorticoids increase transcription of BCKAD subunit genes, transfection studies were performed with BCKAD promoter-luciferase reporter minigenes in LLC-PK(1) cells which do not express gluco-corticoid receptors or LLC-PK(1) cells which express a rat glucocorticoid receptor gene (LLC-PK(1)-GR101). Acidification significantly increased luciferase activity in LLC-PK(1) cells and LLC-PK(1)-GR101 cells transfected with reporter plasmids containing 7.0 kb of E2 subunit or 0.8 kb of E1alpha subunit promoter region, respectively. Glucocorticoids in the form of dexamethasone induced transcription of these minigenes but only in LLC-PK(1) GR101 cells. Using promoter deletion analysis, independent transactivation response elements to acidification or glucocorticoids were localized in the E2 promoter. In summary, catabolic responses to low extracellular pH and glucocorticoids include enhanced expression of genes encoding BCKAD subunits. PMID- 10681645 TI - Effects of acidosis on acute phase protein metabolism in liver cells. AB - Metabolic acidosis has been shown to act as a causative factor in muscle protein breakdown and negative nitrogen balance, as well as in decreased albumin synthesis. Albumin and other acute phase proteins (APP) are mainly synthesized in the liver following induction by interleukins, hormones and other mediators. Acute phase proteins have been shown to be predictors of cardiovascular mortality in the general population and in patients with end stage renal disease (ESRD). Clinical investigation gives evidence that albumin is reduced by acidosis in ESRD patients. The aim of our study was to investigate the role of the liver in acidosis, i.e. the influence of acidosis on metabolic activity and secretion of APP by liver cells (HepG2). Cells were cultured in a medium containing different amounts of bicarbonate. Metabolic activity was significantly diminished when the bicarbonate concentration of the extracellular medium was reduced (86.13+/-1.90% (pH 7.0) vs. 99. 53+/-90% (pH 7.4); p<0.01). While cellular release of negative APP was significantly decreased (albumin: 4.6+/-0.41 (pH 7.0) vs. 7.54+/-0.62 (pH 7.4) [ng/microg protein], p<0.001, transferrin: (0. 78+/-0.08 (pH 7.0) vs. 1.07+/ 0.07 (pH 7.4) [ng/microg protein], p<0. 05), no significant influence of acidosis (pH 7.0) on the positive APP, alpha(1)-acid glycoprotein (AGP) (1.69+/-0.25) (pH 7.0) vs. 1.62+/-0.23 (pH 7.4) [ng/microg protein]), could be shown. Our data indicate that acidosis results in inhibition of liver cell metabolic activity and in reduced secretion of the negative acute phase proteins albumin and transferrin. In contrast, secretion of the positive acute phase protein AGP seems to be unchanged at pH 7.0 as compared to pH 7.4. We conclude that negative and positive APP in liver cells (HepG2) appear to be differently regulated by acidosis. PMID- 10681646 TI - Pathophysiologic glomerulotubular growth factor link. AB - Circumstantial evidence from clinical and pathologic correlations in patients with glomerular diseases and proteinuria suggest that glomerular protein ultrafiltration contributes to tubulointerstitial injury. A series of studies was performed to examine the hypothesis that in rats with adriamycin-induced nephropathy or with diabetic nephropathy (but not in normal rats) high molecular wt. growth factors are ultrafiltered into tubular fluid and act on tubular cells through apical membrane receptors. Analysis of proximal tubular fluid that was collected by nephron micropuncture indicates ultrafiltration of IGF-I, TGF-beta and HGF. Respective receptors are also expressed in apical membranes in some parts of the nephron as examined by immunohistochemistry. In vitro cell culture experiments using proximal tubular fluid obtained from rats with experimental glomerular diseases indicate that ultrafiltered IGF-I may contribute to increased distal tubular Na-absorption. Indirect evidence also suggests that this growth factor may increase the secretion of collagen types I and IV in proximal tubular cells. TGF-beta and HGF cause increased expression and basolateral secretion of MCP-1 in proximal tubular and collecting duct cells. There may be other biologic effects on tubules that are caused by apical exposure to ultrafiltered growth factors. These studies suggest that the glomerular ultrafiltration of bioactive proteins causes or contributes to tubulo-interstitial pathology in glomerular proteinuria. PMID- 10681647 TI - Inflammation nutritional state and outcome in end stage renal disease. AB - Hypoalbuminemia and reduction in lean body mass are potentially a reflection of malnutrition and portend a poor prognosis in patients with end stage renal disease (ESRD). The classic understanding of this relationship has been that ESRD patients receive insufficient dialysis, reduce dietary intake and become malnourished. Inflammation also causes many of the same changes in serum protein composition and in body morphometry as malnutrition does even with adequate calorie and protein intake. It has recently been recognized that this, the acute phase response, occurs with frequency in ESRD patients and that both the physical attributes of malnutrition and reduction in the serum concentration of albumin, transferrin, prealbumin and apolipoprotein A-I all may lack a nutritional base. The serum concentration of the acute phase proteins, C reactive protein (CRP) and serum amyloid A (SAA), as well as the cytokines orchestrating the acute phase response, predict albumin concentration as well as a number of clinically important outcomes: specifically, erythropoietin resistance rejection of renal transplant and survival. ESRD per se does not cause the acute phase response, and indeed may blunt the response to infection. Activation of the acute phase response may be a consequence of the interaction of mononuclear cells with dialysis membranes, especially cuprophane, with endotoxin in dialysate, or represents either clinically evident or obscure infection. Evaluation of the acute phase response by measurement of CRP is advisable in the evaluation of hypoalbuminemia or other stigmata of malnutrition in dialysis patients. PMID- 10681648 TI - Genesis of atherosclerosis in uremic patients. AB - Atherosclerosis significantly contributes to the high incidence of cardiovascular complications and death of uremic patients. Whereas some of the mechanisms that are being discussed in the atherogenesis of chronic renal failure are well established, many others remain hypothetical at present. One of the main reasons is that no long-term, well-controlled studies are available in this area which would have systematically tested the importance of factors such as high blood pressure, via the use of antihypertensive drugs, or disturbances of lipoprotein metabolism, via the use of lipid-lowering medications. Current clinical practice as to the use of the latter has been extensively reviewed in a recent meta analysis by Massy et al. [69]. The majority of studies have been limited to relating hemodynamic or metabolic parameters to parameters of ischemia. Controlled long-term studies are required to reach valid conclusions on the implications of the numerous factors which theoretically may play a role in the atherosclerotic complications of chronic renal failure patients. PMID- 10681649 TI - Apolipoprotein C-III, hypertriglyceridemia and triglyceride-rich lipoproteins in uremia. AB - Apolipoprotein C-III (ApoC-III) plays an important role in the metabolism of triglyceride-rich lipoproteins and is known to be elevated in patients with uremia. To investigate the role of apoC-III in uremic dyslipidemia, we examined apoC-III, triglyceride levels and lipoprotein particles containing both apoB and apoC-III (LP-Bc) in 27 uremic patients prior to dialysis (predialysis), 30 patients on hemodialysis (HD) and 31 patients on peritoneal dialysis (PD). All three groups of patients had elevated levels of plasma apoC-III (20+/-7 mg/dl for predialysis, 18+/-5 for HD and 22+/-8 for PD, compared to 11+/-3 mg/dl for control subjects [p<0/01 for all comparisons]). ApoC-III was positively correlated with plasma triglycerides in PD patients (r = 0.86, p<0.0001), HD patients (r = 0.67, p<0.0001) and predialysis patients (r = 0.60, p<0.001) as well as in all patients combined (r = 0.75, p<0.0001). ApoC-III was also positively correlated with levels of LP-Bc in all three groups of patients, although this correlation was less strong (r = 0.46, p<0.0001 for all patients combined). In predialysis and PD patients, the majority of apoC-III was found in heparin precipitable lipoproteins, whereas the majority of apoC-III in HD patients was found in HDL, indicating less efficient lipolysis in predialysis and PD patients in comparison with HD. These data support the hypothesis that the elevation of apoC-III in uremia can alter the metabolism of triglyceride-rich lipoproteins, leading to an elevation in triglycerides and LP-Bc. Understanding the mechanism(s) of elevated apoC-III in uremia may help to clarify the causes of uremic dyslipidemia. PMID- 10681650 TI - Modification of lipoproteins in uremia: oxidation, glycation and carbamoylation. AB - Lipoprotein modification occurs in uremic patients and in patients with end stage kidney disease under chronic renal replacement therapy. Forms of lipoprotein modification include lipid peroxidation, glycation, and carbamoylation. In this short review, we discuss the presence of these forms of lipoprotein modification and their association with various renal diseases. Methods to analyze lipoprotein modification are introduced, and functional consequences related to vascular and renal function are presented. PMID- 10681651 TI - Molecular genetics of homocysteine metabolism. AB - Recent genetic studies have led to the characterization of molecular determinants contributing to the pathogenesis of hyperhomocysteinemia. In this article we summarize the current insights into the molecular genetics of severe, moderate and mild hyperhomocysteinemia. We will consider deficiencies of the trans sulfuration enzyme cystathionine beta-synthase (gene symbol: CBS), and the disturbances of the remethylation enzymes 5, 10-methylenetetrahydrofolate reductase (gene symbol: MTHFR), methionine synthase (gene symbol: MTR), and the recently identified methionine synthase reductase (gene symbol: MTRR). Furthermore, we will focus on clinically important genetic polymorphisms which are highly prevalent and thus of potential general interest. PMID- 10681652 TI - Homocysteine and chronic renal failure. AB - Homocysteine, a sulfur amino acid, is an important methionine derivative, which has been implicated in the pathogenesis of atherothrombosis. Although only observational, epidemiological studies are available at present, the evidence of an association between hyperhomocysteinemia and increased cardiovascular risk is quite strong and this is confirmed also in a population of chronic renal failure patients. From a biochemical standpoint at least three mechanisms have been summoned so far in order to explain homocysteine toxicity including: oxidation, hypomethylation, and acylation. Proteins are believed to play a crucial role as homocysteine molecular targets. Interference with the functions of several of such macromolecules has been so far described being mediated by any of the above mechanisms. Vitamins may positively influence homocysteine metabolism, thus facilitating the metabolic clearance of this compound. Therefore they are presently considered as potential means for reducing plasma levels of this amino acid and preventing vascular occlusions in hyperhomocysteinemic patients. These compounds, with special regard to folate, are eligible for interventional clinical trials, from which the definitive answer on the role of homocysteine in atherothrombosis is expected. PMID- 10681653 TI - Pathophysiology and clinical importance of hyperhomocysteinemia: clinical intervention studies. AB - Hyperhomocysteinemia is a cardiovascular disease risk factor in the general population and in patients with renal failure. This article summarizes the results of recent total homocysteine-lowering intervention studies in individuals without renal failure and in patients with renal failure. Although almost all of the published studies mainly focused on the total homocysteine-lowering effect and not on cardiovascular disease outcomes of folic acid and vitamin therapy, recent work has gained important insights into this area of developing concern. PMID- 10681654 TI - Regulation of appetite in chronic renal failure. AB - Anorexia, nausea and vomiting in patients with severe renal failure may cause or contribute to development of protein-energy malnutrition, which is associated with increased morbidity and mortality. However, the specific mechanisms that cause appetite suppression in uremia are poorly understood. This review summarizes the general mechanisms by which appetite is regulated. Various factors are discussed that may potentially be involved in appetite suppression in chronic renal failure. PMID- 10681655 TI - Leptin and its clinical implications in chronic renal failure. AB - Leptin, the recently identified ob gene product, regulates food intake and energy expenditure in animal models. Leptin reaches the brain by a saturable transport mechanism and, via direct effects on the hypothalamus, decreases appetite and increases metabolism. Several recent studies have demonstrated markedly elevated serum leptin levels in patients with chronic renal failure (CRF) and it has been speculated that hyperleptinemia may contribute to uremic anorexia and malnutrition. Several factors may influence serum leptin levels in uremia and apart from decreased glomerular filtration rate also body fat mass and plasma insulin levels are important factors that determine serum leptin levels. The possible influence of chronic inflammation on serum leptin levels in CRF need further studies. Patients treated by peritoneal dialysis seem to have higher leptin levels compared to patients treated by hemodialysis. This could be the effect of a marked increase in body fat mass as a consequence of the continuous carbohydrate load. Leptin receptors have by now been identified in several peripheral organs which suggests that leptin besides having central effects also has a pleiotropic action. Indeed, recent findings indicate that besides regulating appetite leptin may play a role in sympathico-activation, insulin metabolism, renal sodium handling and hematopoiesis. PMID- 10681656 TI - Effects of neuropeptide Y on appetite. AB - Neuropeptide Y (NPY) is a polypeptide containing 36 amino acids. Circulating NPY originates predominantly from the sympatho-adrenomedullary nervous system. It has a vasoconstrictive and mitogenic effect on blood vessels and seems to be involved in blood pressure regulation and angiogenesis. NPY is a potent orexigenic agent and is presumed to play a leading role in the regulation of eating behavior. Stimulation of the NPY-ergic arcuate - paraventricular nucleus (ARC-PVN) pathway by exercise, fasting, energy loss (glucosuria) is followed by increased appetite and food intake and increased parasympathetic activity, but suppression of sympathetic activity and energy expenditure. The end result of this process is an increase of energy stores. Activity of the NPY-ergic ARC-PVN pathway is suppressed by leptin - a polypeptide produced by adipocytes. Although functioning of an NPY-leptin feedback was found in rodents, it seems likely that also in man the NPY-leptin axis is involved in the regulation of food intake and energy expenditure. PMID- 10681657 TI - High protein/energy vs. standard protein/energy nutritional regimen in the treatment of malnourished hemodialysis patients. AB - Although malnutrition is frequently encountered in maintenance hemodialysis (MHD) patients, a clear method of treating this complication is still lacking. Failure of nutritional support regimens may be due to inadequate support of dietary needs. Therefore, a high vs. standard or low protein/energy dietary regimen was studied in malnourished MHD patients. A total of 18 malnourished MHD patients selected according to subjective global assessment (SGA)-scores and biochemical indicators of malnutrition (serum albumin <40 g/l, cholesterol <200 mg/dl, prealbumin <30 mg/dl; two out of three) were assigned to three treatment groups: (A: 45 kcal/kg/d and 1.5 g protein/kg/d; B: 35 kcal/kg/d and 1.2 g protein/kg/d; C: spontaneous intake supplemented with 10% of mean protein and energy intake). A and B received food supplements at appropriate dosing to reach the targeted nutritional intake. During 3-month follow-up nutrient intake was assessed by repeated 4-day dietary diaries. Compliance and tolerance was good in each group. Weight gain (1.2+/-0.4 kg) was observed in group A, but not in B and C. Serum albumin levels increased by 1.0+/-0.5 g/l in group A, but not in B and C. Prealbumin and cholesterol levels were unaffected. Weight change correlated with mean dietary energy intake, but not with mean dietary protein intake. We conclude that prescription of 45 kcal/kg/d and 1.5 g protein/kg/d may be necessary to achieve weight gain and improvement of nutritional indices in malnourished MHD pts. Oral food supplements can be used safely and effectively to increase nutrient intake to high levels in these patients. PMID- 10681658 TI - Low protein diets are not needed in chronic renal failure. AB - Low protein diets have been used for a long time in the conservative management of chronic renal failure as they have a beneficial effect in preventing the appearance of symptoms. However, with the exception of the beneficial effect on hyperphosphatemia of the very low protein diets supplemented with ketoacids, they have no proven effects on the other aspects of the uremic syndrome. Moreover, the weight of the evidence suggests that the effect of these diets on preservation of GFR, if any, in patients with nondiabetic renal disease is small and of little clinical relevance. There is very little evidence in the literature of its role in patients with diabetes. The nutritional safety of these diets is still suspect. Patients with chronic renal failure have low energy intakes, which is further reduced when these diets are prescribed. Metabolic studies predict that these patients would be in negative nitrogen balance and in fact, even nutritionally sound, nondiabetic patients enrolled in the Modification of Diet in Renal Disease Study developed subclinical signs of malnutrition. It is possible that the nutritional decline may have been more pronounced on longer duration of follow-up. Finally, these diets are difficult to follow, leading to issues of compliance and exert a great toll on the time of the dietitians. Hence, we conclude that low protein diets are not necessary in chronic renal failure. PMID- 10681659 TI - The potential of intradialytic parenteral nutrition: A review. AB - Malnutrition is common in chronic hemodialysis (CHD) patients and is strongly related to increased morbidity and mortality. Among the various approaches to treat malnutrition in this patient population, intradialytic parenteral nutrition (IDPN) is the treatment of choice for a small but important percentage of malnourished CHD patients. However, the new revised policies relating to IDPN reimbursement by Medicare in the US have made it very difficult to qualify patients for this potentially useful therapy. This restrictive policy was adopted mainly because there are no clear data that support IDPN use or efficacy. Studies to date in the literature do not provide clear documentation of the benefits of IDPN or their cost-effectiveness. The purpose of this review is to critically evaluate studies relating to the use of IDPN as a potential therapy to treat malnutrition in CHD patients and to discuss potential trials to prove its cost effectiveness. PMID- 10681660 TI - Intradialytic parenteral nutrition in malnourished hemodialysis patients. Review of the literature. AB - Malnutrition is a frequent problem of patients on intermittent hemodialysis and is associated with increased morbidity and mortality. Intradialytic parenteral nutrition (IDPN), i.e. intravenous supplementation of mixtures of glucose, amino acids and/or lipids during the hemodialysis session, is one of the therapeutic measures that are applied to correct this malnutrition. To our knowledge only few long-term clinical studies have been undertaken, evaluating the effect of intravenous calorie administration in hemodialysis. Most studies were carried out over a relatively short observation period in small study populations; in several of these studies, no measures were taken to prevent losses of nutrients in the dialysate; adequate control groups are often missing. The authors review the current available literature and conclude that IDPN might have a significant beneficial effect on the nutritional status in malnourished hemodialysis populations. PMID- 10681661 TI - Nutrition impact of peritoneal dialysis solutions. AB - All peritoneal dialysis (PD) solutions are designed to remove toxins and water, normalize the blood electrolyte profile, and provide alkali to help maintain acid base balance. Different formulations, however, may have different effects upon nutrition status. Solutions with 40, as opposed to 35, mEq/l of sodium lactate have been found to promote weight and muscle mass gain and reduce hospitalization in malnourished PD patients. Glucose is varied to produce solutions with different ultrafiltration potential. The glucose absorbed from the PD solution has a protein-sparing effect. The high glucose concentrations necessary for sustained ultrafiltration over a long dialysis dwell, however, often produce appetite suppression and metabolic abnormalities. Solutions formulated with glucose polymers, instead of hypertonic glucose, may provide sustained ultrafiltration over long dwells with lower carbohydrate absorption and perhaps fewer metabolic effects. Amino acids can also be substituted for glucose at relatively low concentrations. A number of studies have shown that amino acids absorbed from the dialysis solution can provide nutritional benefit to malnourished PD patients. PMID- 10681662 TI - Vitamin D analogs: perspectives for treatment. AB - Vitamin D therapy is widely used for the treatment of secondary hyperparathyroidism associated with chronic renal failure. However, administration of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] or its precursor 1alpha(OH)D(3), especially in combination with calcium-based phosphate binders, often produces hypercalcemia. Several vitamin D analogs have been developed that retain the direct suppressive action of 1,25(OH)(2)D(3) on the parathyroid glands but have less calcemic activity. These analogs offer a safer and more effective means of controlling secondary hyperparathyroidism. 22-Oxa-1,25(OH)(2)D(3) (22 oxacalcitriol or OCT), 19-nor-1, 25(OH)(2)D(2) (19-norD(2)) and 1alpha(OH)D(2) have been tested in animal models of uremia and in clinical trials. Intravenous 19-norD(2) and oral 1alpha(OH)D(2) have been approved for use in the United States; OCT is currently under review. The mechanisms by which these analogs exert their selective actions on the parathyroid glands are under investigation. The low calcemic activity of OCT has been attributed to its rapid clearance which prevents sustained effects on intestinal calcium absorption and bone resorption, but still allows a prolonged suppression of PTH gene expression. The selectivity of 19-norD(2) and 1alpha(OH)D(2) is achieved by a distinct mechanism(s). Knowledge of how these compounds exert their selective actions on the parathyroid glands may allow the design of more effective analogs in the future. PMID- 10681663 TI - Transcriptional and post-transcriptional regulation of PTH gene expression by vitamin D, calcium and phosphate. AB - 1,25(OH)(2)D(3) the biologically active metabolite of vitamin D is synthesized in the renal proximal tubules from the hepatic metabolite 25 (OH)D. Lack of 1,25(OH)(2)D(3) is relevant to the pathogenesis of secondary hyperparathyroidism, and 1,25(OH)(2)D(3) itself is used effectively in the management of renal failure patients to prevent secondary hyperparathyroidism. The scientific basis of this therapy is the finding that 1,25(OH)(2)D(3) potently decreases PTH gene transcription both in vitro and in vivo. PMID- 10681664 TI - Nonclassical effects of 1alpha,25-dihydroxyvitamin D(3) and its analogs. AB - The activated form of vitamin D(3), 1alpha,25(OH)(2)D(3), not only plays a central role in bone and calcium metabolism but has also potent antiproliferative and prodifferentiating effects. Moreover, the combined presence of 25(OH)D(3) 1alpha-hydroxylase as well as the vitamin D receptor in several tissues introduced the idea of a paracrine role for 1alpha,25(OH)(2)D(3). By introducing chemical modifications into the flexible parent molecule 1alpha,25(OH)(2)D(3), a whole generation of vitamin D analogs was created. Due to a clear dissociation of the antiproliferative and prodifferentiating effects from calcemic effects, these analogs can be used not only for the treatment of bone disorders but also for non classical applications. PMID- 10681665 TI - Vitamin replacement therapy in renal failure patients. AB - Renal failure patients require vitamin replacement therapy that addresses the specialized needs of renal failure. Four factors including restricted diet, uremic toxins, drug-nutrient interactions, and in ESRD, the dialysis process, affect the normal absorption, retention and activity of necessary micronutrients which support all aspects of carbohydrate, protein, lipid and nucleic acid metabolism. Studies have shown that the typical renal failure diet is low in B vitamins, that uremic factors affect folate and pyridoxine activities and that many B vitamins are lost on dialysis at a rate greater than are lost with normal urinary excretion. In addition, retention of vitamin A or inappropriately high supplementation of vitamin C may cause toxicities which exacerbate existing pathologies. Further, emerging research suggests some vitamins such as folic acid and pyridoxine, if provided in higher than normal amounts, may have an impact on reducing the risk of some aspects of renal cardiovascular disease. It is therefore important to supplement some vitamins, and use restraint in the supplementation of others. It is clear that renal failure patients, including predialysis, ESRD and transplant patients need specialized supplementation that meets the requirements of disease and its management. PMID- 10681666 TI - Influence of water and sodium diuresis and furosemide on urinary excretion of vitamin B(6), oxalic acid and vitamin C in chronic renal failure. AB - Urinary excretion of vitamin B(6), oxalic acid and vitamin C was investigated in 15 healthy subjects during maximal water diuresis and in the group of 12 patients in polyuric stage of chronic renal failure without dialysis treatment receiving a diet containing high sodium chloride (15g/day). Urinary excretions of the same parameters were investigated in another group of 15 patients in polyuric stage of chronic renal failure without dialysis treatment after i.v. administration of 20 mg furosemide. Urinary excretion of vitamin B(6), oxalic acid and vitamin C significantly increased during maximal water diuresis while during high intake of sodium chloride the urinary excretions of these substances were not affected. The results suggest that urinary excretion of vitamin B(6), oxalic acid and vitamin C depends on the urinary excretion of water. Intravenous administration of 20 mg furosemide led to an increase of urinary excretion of vitamin B(6), oxalic acid and vitamin C in patients with chronic renal failure. The increased urinary excretion of vitamin B(6) and vitamin C is a new negative side effect of furosemide and increased urinary excretion of oxalic acid is a new positive side effect in patients with chronic renal failure. PMID- 10681667 TI - Metabolic adjuvants to erythropoietin therapy. AB - It is possible to potentiate the response to epoietin by co-administering other agents. In some instances, this response is seen when there is a deficiency of a certain substance. In other cases, administration of an adjuvant such as intravenous iron, vitamin D, L-carnitine, or androgens can enhance the response to epoietin when given as a surfeit. With most of these agents, with perhaps the exception of intravenous iron, further research is required to determine the exact role they may play in clinical practice. As long as the cost of epoietin therapy remains fairly high, the challenge will continue as to the best way of optimizing its effect, and we can look forward to new developments in this expanding area of research. PMID- 10681668 TI - Preventive effects of an oral sorbent on nephropathy in rats. AB - Circulating uremic substances are thought to be involved in the progression of chronic renal failure (CRF). An oral adsorbent AST-120 (Kremezin) is effective in removing circulating uremic toxins from the gastrointestinal tract, and retards the progression of CRF. AST-120 is widely used as an approved drug in Japan for the treatment of undialyzed uremic patients to delay the progression of CRF. AST 120 attenuates the progression of glomerular sclerosis and interstitial fibrosis in a variety of experimental rat models of CRF. However, the mechanism by which AST-120 delays the progression of CRF had not been clear. We have demonstrated that indoxyl sulfate, a dietary protein metabolite, is a circulating uremic toxin stimulating glomerular sclerosis and interstitial fibrosis, and that AST-120 decreases the serum and urine levels of indoxyl sulfate by adsorbing its precursor, indole, in the intestine. The administration of indoxyl sulfate to uremic rats stimulated the expression of transforming growth factor (TGF)-beta1, tissue inhibitor of metalloproteinase (TIMP)-1 and pro-alpha1(I)collagen in the kidneys. Further, the administration of AST-120 to uremic rats reduced the extent of glomerular sclerosis and interstitial fibrosis as well as the renal expression of TGF-beta1 and TIMP-1, by reducing the serum and urine levels of indoxyl sulfate. We propose the protein metabolite hypothesis that endogenous protein metabolites such as indoxyl sulfate play an important role in the progression of CRF, and that AST-120 is effective in retarding the progression of CRF by removing these protein metabolites through intestinal absorption. PMID- 10681669 TI - Liver cell reactive components in peritoneal dialysis fluids. AB - Metabolic changes in peritoneal dialysis (PD) patients are an important aspect concerning long-term outcome. Liver plays the main role in regulating metabolism. The effects of peritoneal dialysis fluids (PDF) on liver cell function are scarcely investigated. Therefore, we investigated the effects of PDF, different in some components, on liver cell metabolism in vitro. Metabolic activity (MTT), cell integrity (LDH release), proliferation (BrdU incorporation) and synthesis of albumin and transferrin are measured by incubating HepG2 cells for 3 h and 24 h with six different PDFs: (a) lactate-buffered, pH5.5: PDF I (1.5% gluc.); PDF II (4.5% gluc. ); (b) bicarbonate-buffered, pH7.4: PDF III (1.5% gluc.), PDF IV (4. 5% gluc.); (c) amino acid-based solutions, pH 7.4: PDF V (low AA level) and PDF VI (high AA level). Metabolic activity of bicarbonate-treated cells is greatly enhanced in comparison to lactate-buffered PDFs. These findings are confirmed by proliferation data. Synthesis of albumin and transferrin is significantly enhanced by amino acid-based solutions. Our data demonstrate, that lactate buffered PDF impair liver cells much stronger than bicarbonate-buffered PDF. pH is the parameter which contributes to cytotoxicity and impaired metabolism to a major extent. In contrast to glucose-containing solutions, amino acid-based PDF stimulate protein synthesis in liver cells. PMID- 10681670 TI - Advanced glycation end-product levels in subtotally nephrectomized rats: beneficial effects of angiotensin II receptor 1 antagonist losartan. AB - The angiotensin II receptor 1 antagonist losartan (L) inhibited the advanced glycated end-products (AGEs) induced expression of transforming growth factor beta(1) in in vitro experiments performed on renal tubuloepithelial cells. To test the pathophysiological importance of these findings, the possible link between serum AGEs levels and angiotensin system was investigated in the model of normotensive subtotally nephrectomized rats(4/6-NX). Concentration of AGEs in serum of placebo administered 4/6-NX rats (n = 7, 1.09+/-0.09 U/l) increased slightly in comparison with sham-operated healthy controls (CTRL, n = 8, 0.94+/ 0.10 U/l, p<0.02) as measured by competitive ELISA. Treatment of 4/6-NX rats with L over 12 weeks ameliorated the rise in serum AGEs concentration (1.00+/-0.12 U/l, n = 15 <0.005) almost to the level observed for CTRL. This effect was further corroborated by the observation, that the impaired renal excretion of AGEs in 4/6-NX-placebo rats (0.07+/-0.02 U/micromol creatinine) was significantly restored by L (0.09+/-0.02 U/micromol creatinine, <0.009) and resembled that of the CTRL (0.10+/-0.03 U/micromol creatinine). Administration of L to 4/6-NX rats significantly improved renal function as evaluated by a smaller rise in serum creatinine and urea concentration. In spite of the improvement in renal function, there were no differences in concentrations of transforming growth factor beta(1) in serum and in urine among the two groups. These effects were independent of blood pressure. Our data give first evidence, that long-term treatment with angiotensin II receptor 1 antagonist may exert salutary effects on AGEs levels in the rat remnant kidney model, probably due to improved renal function. PMID- 10681671 TI - Nitric oxide/L-arginine in uremia. AB - Nitric oxide (NO), a gaseous free radical derived from L-arginine, is a potent modulator of vascular tone and platelet functions. A number of recent studies, both in the experimental model of renal mass reduction (RMR) in rats and in uremic patients, have raised the hypothesis that abnormalities of NO synthetic pathway could have a key role in mediating the complex hemodynamic and hemostatic disorders associated to the progression of renal disease. Thus, kidneys from rats with RMR produce less NO than normal rats and NO generation negatively correlates with markers of renal damage. The abnormality is due to a strong defect of inducible NO synthase (iNOS) content in the kidney. Recent in vitro and in vivo data have raised the possibility that excessive renal synthesis of the potent vasoconstrictor and promitogenic peptide endothelin-1 (ET-1) is a major determinant for progressive iNOS loss in the kidney of RMR rats. In contrast, uremia is associated with excessive systemic NO release, both in experimental model and in human beings. In the systemic circulation of uremic rats, as well as uremic patients, NO is formed in excessive amounts. Possible cause of the increased NO levels is higher release from systemic vessels due to the augmented expression of both iNOS and endothelial NOS. A putative cause for excessive NO production in uremia can be guanidinosuccinate, an uremic toxin that accumulates in the circulation of uremic patients and upregulates NO synthesis from cultured endothelial cells. Upregulation of systemic NO synthesis might be a defense mechanism against hypertension of uremia. On the other hand, more NO available to circulating cells may sustain the bleeding tendency, a well-known complication of uremia. PMID- 10681672 TI - Long, slow dialysis. AB - Long slow hemodialysis (3 x 8 hours/week) has been used in Tassin for 30 years without significant change in the method. It provides excellent results in terms of morbidity and mortality. The better survival than usually reported on shorter dialysis is mainly due to lower cardiovascular mortality. The nutritional state of the patient is good, as well as the correction of anemia with low doses of EPO. But the main feature concerns blood pressure; hypertension is very well controlled without need for antihypertensive medications. The gentle ultrafiltration provided by a long session time associated with a low salt diet and a moderate interdialytic weight gain allows for normalization of the extracellular fluid space in most patients (dry weight) without important intradialytic morbidity. This low salt diet has paradoxically been forgotten in recent years while shortened dialysis time renders it more necessary than ever. PMID- 10681673 TI - Nutritional status assessment and body composition analysis in pre-end stage renal disease patients. AB - Malnutrition is a known risk factor for survival in renal failure patients. Of concern, a significant degree of malnutrition may develop in the predialysis period due to dietary restrictions and uremia. To further define this issue, we evaluated 25 predialysis patients using serum chemistries, body mass index (BMI), fat free mass (FFM), body cell mass (BCM), and protein appearance rate (PAR) as surrogates of nutritional status and compared their results to those obtained in established hemodialysis patients and recipients of living donor renal allografts during a nine-month observation period. Pre- dialysis patients had significantly (p<0.0001) higher body weight (28%), body mass index (26%), body cell mass (17%) and fat free mass (15%) than hemodialysis and transplant patients. Intracellular water content was similar in all groups. As many patients do not start dialysis until clearance values fall below 10 ml/min, it is possible that greater tissue mass losses occur in the weeks preceding initiation of dialytic therapy. Why renal transplant recipients fail to increase tissue mass may relate to the catabolic effects of immunosuppression. We conclude that the early stages of pre end stage renal disease are associated with relatively good preservation of body cell mass. PMID- 10681674 TI - Bioelectrical impedance analysis in dialysis patients. AB - Identification of malnutrition is imperative in chronic dialysis patients. Bioelectrical impedance (BIA) is a noninvasive method to measure body composition and estimate total body water (TBW), lean body mass (LBM) and body cell mass (BCM). Studies suggest BIA has good reliability as compared to other accepted methods of body composition analysis. Preliminary data also suggest that BIA derived parameters (reactance and phase angle) predict clinical outcome in chronic hemodialysis patients. Overall, BIA is a promising nutritional assessment tool to monitor health status, long-term follow-up, tailor nutrition support, and detect early subtle losses of LBM in chronic dialysis patients. PMID- 10681675 TI - Cranial bone fixation in infants and children. AB - A technique is described for fixing cranial bone flaps with absorbable sutures and bone shims. This technique is a cost-efficient method of rapidly achieving rigid structural stability with excellent cosmetic results. The kerf created by the craniotome is bridged at multiple sites by bone chips harvested from the inner edge of the bone flap. Solid bony union is documented on histopathological analysis. The technique requires no special tools and avoids the problems associated with the use of plates, screws and wire. PMID- 10681676 TI - Experiences with cranial neuronavigation in pediatric neurosurgery. AB - Frameless, infrared-based neuronavigation (Surgical Microscope Navigator and Surgical Tool Navigator) was applied to intracranial neurosurgery in 22 children and infants (age range 7 months to 16 years). Predominant diagnoses were tumor and hydrocephalus (including multiloculated forms); the surgical procedures were open microneurosurgery in 10 cases and neuroendoscopy in 12 patients. There were no principal technical problems even in very young infants. Surgical and clinical results were satisfying in all cases. Neuronavigation proved to be particularly helpful in targeting deep-seated processes and in guiding neuroendoscopy in cases of multiloculated hydrocephalus or small ventricles. PMID- 10681677 TI - Endoscopic approach to noncommunicating fluid spaces in the shunted patient. AB - INTRODUCTION: It is well known that shunted hydrocephalic patients can, over time, develop entrapped ventricles or cystic spaces not in communication with the remaining ventricles. This situation has traditionally been treated with placement of an additional catheter or shunt system in the noncommunicating fluid space. With the advent of minimally invasive endoscopic techniques, it has become possible to fenestrate trapped fluid spaces into the shunted ventricular system, thus preventing the need for additional catheters. METHODS: Fifteen shunted patients presented with noncommunicating fluid spaces over a 4-year period at our two institutions. We attempted fenestration procedures in 14 of those patients. The various procedures included: septum pellucidum fenestration, cyst fenestration, third ventriculocisternostomy and a combination of the above. RESULTS: Thirteen of 16 (81%) endoscopic fenestrations successfully prevented the need for a complicated shunting system. There were 3 technical failures (3/16, 19%) in which an additional catheter was added to the existing shunt system at the time of the endoscopic procedure. There were no adverse neurologic effects from the endoscopic procedures; however, in 4 of the 14 patients (29%) a shunt revision was performed between 2 and 30 days following the fenestration. CONCLUSIONS: These results show that the endoscopic approach to establishing communication between noncommunicating CSF spaces in the shunted patient is safe and efficacious in preventing the need for additional CSF catheters. Though we encountered failures, the majority of cases are technically feasible. In these patients early shunt malfunction may occur, most likely due to intraoperative bleeding and will require shunt revision. PMID- 10681678 TI - Thalamic glioblastoma with cerebrospinal fluid dissemination in the peritoneal cavity. AB - Glioblastoma multiforme is one of the commonest primary malignant tumours of the brain with rare incidence of extracranial metastases. Systemic dissemination via the CSF or CSF diversionary shunt procedures is also rare. The reported 9-year old child was a case of thalamic glioblastoma with hydrocephalus who underwent biventriculoperitoneal shunting before tumour decompression and radiotherapy. The child developed incapacitating ascites 8 months following surgical decompression and 9 months after the shunt diversion which was found to be caused by CSF dissemination of the glioblastoma via the ventriculoperitoneal shunt. The child ultimately succumbed to his disease. PMID- 10681679 TI - Successful use of intracavitary bleomycin for low-grade astrocytoma tumor cyst. AB - We report successful use of bleomycin in a low-grade astrocytoma tumor cyst of the tectal plate. A 6-year-old male underwent subtotal resection of a low-grade astrocytoma of the tectal plate followed by chemotherapy and proton beam radiation at age 2 and a half. Despite resolution of the solid portion of the tumor, serial MRI showed enlargement of a bilobar tumor cyst 3 years after the original diagnosis. The patient developed progressive ataxia, short-term memory loss and dysconjugate gaze. Following stereotactic placement of an Ommaya reservoir into the cyst, Isovue contrast and CT scan were used to confirm the integrity of the cyst. Five consecutive daily doses of 3.0 mg of bleomycin were instilled into the cyst after removal of cyst fluid. The therapy was well tolerated in the outpatient setting, and the clinical findings resolved. Subsequent CT and MRI at 4 months and 2 years after bleomycin confirmed no recurrence of the tumor or cyst. PMID- 10681680 TI - Inflicted traumatic brain injury: relationship of developmental outcome to severity of injury. AB - Inflicted traumatic brain injury (TBI) is a frequent consequence of physical child abuse in infants and children. Twenty-eight children who were 2-42 months of age when hospitalized for moderate to severe TBI were enrolled in a prospective, longitudinal study of neurobehavioral outcome following acquired brain injury. Relative to a comparison group, the children with inflicted TBI had significant deficits in cognitive, motor and behavioral domains when assessed with the Bayley Scales of Infant Development-II 1 and 3 months after the injury. Nearly half of the injured children showed persisting deficits in attention/arousal, emotional regulation and motor coordination. Greater injury severity, as indicated by lower coma scale scores, longer periods of unconsciousness and the presence of edema/cerebral infarctions was associated with poorer outcomes in all domains. PMID- 10681681 TI - Skull base growth in childhood. AB - While studying skull base changes in craniosynostosis, it became apparent that there is a lack of reference studies quantifying the changes of three-dimensional (3D) parameters of the normal skull base throughout childhood. Using advanced 3D visualisation techniques, 34 points of the skull base were identified on MRI scans of 66 normal children, aged 1 month to 15 years. Several distances and angles between the various landmarks were measured in an attempt to quantify the growth of skull fossae with age. Two main growth periods were observed: before and after the first 5 years of life. Most change occurred in the first period. Anatomical regional differences were identified between the two sexes. During the first 5 years of life, the anterior fossa showed rapid growth rate with respect to its anterior projection in males, whereas in the females there was a more concentric growth pattern. The body of the sphenoid bone and the middle fossa showed a rapid growth rate in both sexes which was greater in the females. The posterior fossa showed a concentric pattern of growth in both sexes with a greater growth rate in the females. These findings provide new insight into the pattern of growth of the various parts of the skull base and can be used for comparative study of deformities that affect such growth. PMID- 10681682 TI - Late shunt infections. AB - We reviewed the records of the 957 shunt-related operations performed at Cardinal Glennon Children's Hospital over a 10-year period. During that time, 94 shunt infections were recognized. Eight of the infections occurred more than 9 months postoperatively. These differed from early infections in two ways: (1) Staphylococcus aureus was not found to be a pathogen in any late infection. (2) Abdominal pseudocysts were much more frequently found in patients with late developing infections. In addition, the pathogens involved and the temporal distribution of cases suggest most infections that occur more than 9 months postoperatively are more likely caused by secondary bacterial seeding than by bacterial inoculation at the time of operation. PMID- 10681683 TI - Abdominal pseudocysts complicating CSF shunting in infants and children. Report of 18 cases. AB - Abdominal pseudocysts (APC) are rare complications of ventriculoperitoneal shunting for hydrocephalus. The authors studied retrospectively a series of 18 pediatric patients with APC. Signs and symptoms of shunt dysfunction were observed in 15 (83.3%), abdominal complaints in 10 (55.5%) and fever in 6 (33.3%). Prior to the diagnosis of APC, 2 patients suffered exploratory laparotomies due to important abdominal signs and symptoms. Ultrasonography was diagnostic in all cases and proved to be the method of choice in the evaluation of APC. Our series suggest that APC are strongly related to hardware infection and in some cases can result from a previous shunt infection not completely cured. The bacteriological examination of the tip of the peritoneal catheter was a reliable indicator of infection. According to our data, the best treatment seems to be the removal of the shunt system and the insertion of an external ventricular drainage. In our experience, almost half of the patients had a ventriculoatrial shunt replacing the peritoneal shunt at the end of the treatment. PMID- 10681684 TI - Vein of galen malformation. PMID- 10681685 TI - Radiologic and histopathologic changes after Gamma Knife radiosurgery for acoustic schwannoma. AB - Gamma knife radiosurgery (GKRS) is a widely used treatment option for acoustic schwannomas, 3 cm in diameter or less. Between May 1990 and February 1998, 102 acoustic tumors in 101 patients were treated with GKRS. There are 77 patients with a follow-up period of more than six months (mean 55, range 7 to 90 months). Seventy (91%) of these tumors have remained unchanged or reduced in volume. After GKRS there was an increase in volume in seven cases. In four the volume increase affected solid tumour. Among these, three patients were in stable condition and are being observed. One of these patients developed brain stem compression symptoms and was operated. In another three cases, cysts with multiple septa developed medial to the tumor and compressed the brain stem and fourth ventricle, thus necessitating post-GKRS surgery. In these three patients, MRI had shown loss of central contrast enhancement followed by its return. Histological findings at surgery before and after GKRS were compared for these four tumours. In spite of the MRI changes, there were no definite histological findings after GKRS which could be attributed to radiation induced changes. The development of cysts occurred after the treatment of larger tumors. PMID- 10681686 TI - Gamma Knife radiosurgery relative to microsurgery: epilepsy. AB - There is a strong rationale for investigation of the role of Gamma Knife radiosurgery in the treatment of medically intractable epilepsy. To explore this potential application, the well established and highly successful current outcomes associated with microsurgical treatment were reviewed, and include for temporal resections seizure-free results in 65-70% of patients, with permanent morbidity of less than 5% and mortality less than 1%. Advantages of open surgery include the opportunity to conduct electrocorticography and functional mapping, excellent visualization enabling an assured and discrete line of resection, freedom in general from volume constraints, and immediate efficacy. The preliminary evidence in support of radiosurgical treatment includes several series of patients with epilepsy associated with arteriovenous malformation, tumor, or cavernous anginoma in which results approaching those of surgery have been achieved. In a series of 15 patients with mesial temporal sclerosis, all but one patient with follow-up of at least one year are seizure-free; morbidity has been limited to one asymptomatic field defect. A series of ten patients with epilepsy associated with hypothalamic hamartoma has achieved seizure improvement in six of eight patients with at least one year follow-up, and no major morbidity has been seen. Radiosurgical treatment for intractable epilepsy appears to be effective in a majority of patients and can be performed safely and efficiently. Early experimental evidence suggests that seizure control might in some instances be achievable without ablation of the pathological target. PMID- 10681687 TI - Gamma Knife radiosurgery for epilepsy associated with cavernous hemangiomas: a retrospective study of 49 cases. AB - A retrospective multicenter study was performed to evaluate the effectiveness of Gamma Knife radiosurgery (GKRS) in the treatment of drug resistant epilepsy associated with cavernous hemangiomas (CH). The mean duration of epilepsy before GKRS was 7.5 ? 9.3 years. The mean frequency of seizures was 6.9 ? 14/month. The mean marginal dose was 19.2 ? 4.4 Gy (range 11.3 to 36 Gy). The mean follow up was 23.7 ? 13 months. At the most recent follow-up examination, 26 (53%) patients were seizure-free (Engel s class I) including 24 in class IA (49%) and two (4%) patients with occasional auras (class IB, 4%). A highly significant decrease in the number of seizures was achieved in 10 (20%) patients, which is class lIB. The remaining 13 (2.6%) patients showed little or no improvement. A medial temporal location was associated with a higher risk of failure. In contrast, all patients with central region CH were seizure free. Two severe but transient complications were observed. There was hemorrhage in one patient and another patient suffered from radio-induced edema with transient aphasia. This series is the first demonstrating that GKRS can be used safely and efficiently to treat epilepsy associated to CH. Seizure control can be reached when a good electro-clinical correlation exists between CH location and epileptogenic zone. Our findings suggest that GKRS can be used to treat epilepsy for CH located in highly functional areas, particularly the central region. PMID- 10681688 TI - The effects of stereotactic radiosurgery on secondary facial pain. AB - Twenty-seven patients with tumor-related secondary facial pain were treated by stereotactic radiosurgical procedures between November 1991 and October 1998. They had 14 meningiomas, 11 schwannomas (one trigeminal, 10 vestibular), one nasopharyngeal cancer and one chordoma. The mean maximum dose administered was 26.4 Gy (range 16 to 35 Gy) and the margin of the tumor was encompassed within the 45 to 90% isodose. The patients were analyzed based on their pain relief with a mean follow-up duration of 32.1 months. In 24 patients (85.7%), there was initial pain improvement after radiosurgery, but half had recurrent pain. A pain response was obtained in 12 cases (pain response rate = 42.9%), five were pain free and seven had pain reduction. On the follow-up MRI, a decrease in tumor volume of more than 20% of the preoperative volume occurred in 14 of 25 cases. The mean time interval to initial pain improvement (10.3 months) and pain response (5.7 months) were shorter than for a decrease in tumor volume (18.6 months). Tumor-related secondary facial pain was less responsive to stereotactic radiosurgery than idiopathic trigeminal neuralgia. It would seem that the mechanism of pain relief in radiosurgery is not only trigeminal root decompression secondary to tumor volume reduction, but also other mechanisms involving inactivation of abnormal electrical transmission may be involved. PMID- 10681689 TI - Brain tumors with cysts treated with Gamma Knife radiosurgery: is microsurgery indicated? AB - Cysts, which are not uncommonly associated with brain tumors, may be responsible for neurological dysfunction. Stereotactic aspiration of such lesions can lead to clinical improvement, but cyst recurrence is common and multiple aspirations may be necessary. Thirteen cases of brain tumors with cystic components were treated by radiosurgery with follow-up of 5-29 months (median 14 months). The tumor diagnoses were three cystic craniopharyngiomas, two brain stem cystic astrocytomas, two cystic cerebellar astrocytomas, one cerebellar hemangioblastoma, one ganglioglioma, one fourth ventricle tumor, one cerebellopontine angle pilocytic astrocytoma, one metastasis from lung cancer and one glioblastoma. The dose at the tumor margin ranged between 10 and 20.5 Gy (mean 15.5 Gy) and the maximum dose ranged between 18 and 45 Gy (mean 32.3 Gy). In 11 of these cases the cystic component recurred in spite of a decrease in the size of the solid tumor component. An Ommaya reservoir was inserted in six cases, stereotactic aspiration was performed in two cases, microsurgery was undertaken in two cases after 2-8 months (mean 4.8 months) and one patient refused further treatment. Multiple aspirations through the Ommaya reservoir were performed in the outpatients on the two patients who required them. It may be appropriate to be cautious in advising radiosurgery for intracranial tumors with a significant cystic component. Microsurgery if possible may be preferable in this situation. PMID- 10681690 TI - Paradoxical labeling of radiosurgically treated quiescent tumors with Ki67, a marker of cellular proliferation. AB - Ki67 (also known as MIB) is a monoclonal antibody staining agent, which is routinely used as a marker of cellular proliferation. It is used to evaluate the proliferative potential of malignant tumors, since it is thought to stain only those cells undergoing active division. The paradoxical elevation of the Ki67 labeling index observed in radiosurgically treated benign and malignant tumors is reported. Ten patients, who had previously undergone either linac or Gamma Knife radiosurgery, underwent surgical resection for a radiographically quiescent tumor. One patient underwent autopsy after dying from complications of radiosurgery. All were thought to be suffering from adverse radiation effect (ARE) and were refractory to conservative management. None were thought to have a recurrent tumor. All of the resected tumors were subjected to analysis with Ki67 staining. Despite the radiographic stability of the tumors, all manifested significantly elevated populations of cells exhibiting positivity for Ki67 antigen. This staining pattern would suggest a significant proliferative potential, which did not match the observed clinical course. The literature is reviewed and possible mechanisms to explain the paradoxical findings are presented. PMID- 10681691 TI - Radiosurgery of glomus tumors: midterm results. AB - Glomus tumors (GT) of the skull-base present a complex surgical challenge due to their delicate localization and specific vascular supply. This study is designed to evaluate the role of stereotactic radiosurgery in the treatment of GT. Thirteen patients with GT have been treated with the Gamma Knife. Radiosurgery was performed because of recurrences after surgical removal in six patients. Histology was not available in seven patients, thus, diagnosis was made from neuroradiological features only. Two of them had partial embolization before Gamma Knife treatment. Clinical and morphological data were collected from 11 patients, who had a representative follow-up of at least 12 months. Mean follow up was 42 months (range 14 to 72 months). Within the follow-up period there was no tumor progression and no clinical deterioration in any of the patients. 64% of the patients had an improvement of their symptoms, and in 36% the volume of the lesion decreased in size. There was no radiation-related morbidity. It is suggested that radiosurgery seems to be safe and effective in the treatment of GT. PMID- 10681692 TI - A comparison of single fraction radiosurgery tumor control and toxicity in the treatment of basal and nonbasal meningiomas. AB - Between July 1993 and October 1997, 107 patients with 118 meningiomas were treated with Gamma Knife radiosurgery (GKRS). The most frequent site of tumor origin was the skull base (54%). The mean tumor diameter and volume were 2.5 cm and 9.4 cm3, respectively. The mean dose to the tumor periphery was 17 Gy, prescribed to a mean iso-dose of 47%. At a mean follow-up of 28 months, tumor control for basal and nonbasal meningiomas was 80%. Deteriorating peritumoral edema associated with symptoms was observed in 1 of 49 (2%) skull-base tumors and in 4 of 39 (10%) non-basal tumors, without associated tumor growth. (p=0.l5 and 0.234 respectively, z-test). Stereotactic radiosurgery can achieve acceptable tumor control with low morbidity in the treatment of most meningiomas. However, when the tumor is nonbasal, the potential morbidity from peritumoral edema should be recognized and other treatment options considered, such as adjuvant surgery, partial fractionated irradiation or stereotactic radiotherapy. PMID- 10681693 TI - Low-dose radiosurgery for meningiomas. AB - Gamma Knife radiosurgery is an effective treatment for meningiomas. However, it may be difficult to deliver what is currently considered an optimal dose, especially if the tumor is large or adjacent to critical structures. Eleven cases are presented with a follow-up of more than 12 months where the margin dose did not exceed 10 Gy. The mean age was 48.8 years. The mean follow-up period was 35.7 months (range 21 to 57 months). The mean volume of the tumors was 9.4 cm3 (range 1.6 to 28.9 cm3). The margin dose was less than 10 Gy in all 11 cases, due to a large volume in two cases. Four tumors were close to the visual pathways. Five tumors were close to the brain stem. Imaging follow-up showed that four tumors had shrunk after radiosurgery. The remaining seven cases remained unchanged. There was no tumor growth after radiosurgery. A transient oculomotor palsy was observed in two cases after radiosurgery. It is suggested that Gamma Knife radiosurgery using lower dosage than usual is one of the options for the treatment of meningioma. PMID- 10681694 TI - Gamma Knife radiosurgery for metastatic brain tumor: the usefulness of repeated Gamma Knife radiosurgery for recurrent cases. AB - Forty-one patients with a total of 193 metastatic brain tumors were treated because of new lesions or in a few cases because of tumor regrowth. The management involved two to four treatments with Gamma Knife radiosurgery (GKRS). The overall median survival was 15 months. The local control rate of tumors was 89% for the first treatment and 93% for repeated treatments. Symptomatic radiation-induced edema appeared in only two cases (4.9%). Patients with metastatic brain tumors who developed new lesions after GKRS were treated with GKRS alone for subsequent lesions. It was possible to control tumors for a long period with minimal side effects. As a result, we were able to maintain the quality of life of these patients, but, the effect of GKRS in controlling tumor regrowth seemed temporary. PMID- 10681695 TI - Gamma Knife radiosurgery for skull base metastasis and invasion. AB - We treated 18 patients with skull base metastasis or invasion using Gamma Knife radiosurgery. Seven of these patients had invasive nasopharyngeal cancer and 11 had distant metastasis from other cancers. The mean diameter of tumors was 22-46 mm (median 32.1 mm) and the radiation dose to the tumor margin was 12-23 Gy (median 16.2 Gy). The median follow-up period was 10.5 months. Clinical symptoms were improved in 61% of the patients after treatment and tumor control was obtained in 67% of cases at their final followup. Radiation injury occurred in only one patient (6%) who had received previous radiotherapy to the same region. Gamma Knife radiosurgery is a useful therapeutic option for the treatment of skull base metastasis and invasion, either as a secondary treatment for recurrence after previous radiotherapy or as a primary treatment. PMID- 10681696 TI - A six year experience with the postoperative radiosurgical management of pituitary adenomas. AB - Since April 1992, 73 consecutive patients with pituitary adenomas were treated with radiosurgery. There were 31 hormonally inactive adenomas and 42 hormonally active adenomas. All but three patients had been subjected to one or more surgical procedures prior to radiosurgery. Three patients had received fractionated radiotherapy. In the inactive adenoma group, the mean target volume was 4.4 ? 3 cm3 and the mean prescription dose was 13.8?1.5 Gy. In the prolactinoma patients, the mean target volume was 6.7 ? 9 cm3 and the mean prescription dose was 14.2 ?4 Gy. In the acromegalic patients, the mean target volume was 2.9?2.5 cm3 and the mean prescription dose was 16?4 Gy. ACTH secreting adenomas had a mean target volume of 3.6 ? 5.5 cm3 with a mean prescription dose of 17 ?4.8 Gy. The mean follow-up time was 28.9 ? 21.5 months. Follow-up data was available in 83.6% of the patients. Tumor control was achieved in 98.3% and the endocrinological cure rate was 57%. Pituitary function deteriorated in 19.2%. No patient suffered from radiation induced visual damage. It would seem that postoperative radiosurgery for residual or recurrent pituitary adenomas may be a safe technique that can increase the frequency of therapeutic success. PMID- 10681697 TI - Gamma Knife radiosurgery for functioning pituitary adenomas. AB - Stereotactic radiosurgery has been an important treatment modality in the treatment of pituitary adenomas. However, it has the disadvantage of a delayed effect on hormonal normalization compared with microsurgical resection of functioning pituitary adenomas. To define the efficacy of radiosurgery in the treatment of functioning pituitary adenomas, 37 cases with a mean follow-up duration of 26.9 months were analyzed. There were 18 prolactinomas, 11 cases with acromegaly, and 8 cases with Cushing s disease. The mean maximum dose was 54.8 Gy. The tumor margin was encompassed within the 50 to 90% isodose. The level of serum prolactin, growth hormone, and 24-hour urine free cortisol were evaluated for hormonal follow-up according to the relevant endocrinopathy. There was 35.1% hormonal normalization and an 81.8% decline in hormone levels to below 50% of the preoperative value (hormonal response). Hormonal normalization was obtained in 13 patients (mean latency = 22 months). A hormonal response was seen in 30 patients (mean latency = 7.6 months). The maximum dose and tumor volume included in the prescription isodose were significantly correlated with the latency period from radiosurgery to hormonal normalization. These results suggest that early hormonal normalization can best be achieved by a high maximum dose (at least 55 Gy) and broad coverage of the target tumor volume within the prescription dose thereby increasing the integral dose. PMID- 10681698 TI - Gamma Knife radiosurgery for pituitary adenomas. AB - Ninety-two patients with pituitary adenomas have been treated during the last 5 years. Sixty-three of these patients had more than 6 months follow-up, and they form the basis of this report. Eighteen had non-functioning adenomas (NFA), and 36 had functioning adenomas (FA). The mean marginal dose was 22.5 Gy (NFA 19.5 Gy, FA 23.9 Gy). Control of tumor growth was achieved in 92%. A significant decrease of excessive hormone production was seen in 75.6%, and the endocrinopathy normalization rate was 26.7%. Post-radiosurgical complications were seen in 4.7%. PMID- 10681699 TI - Gamma Knife radiosurgery for functioning pituitary microadenoma. AB - Transsphenoidal microsurgery remains the treatment of choice for pituitary microadenomas One hundred and six patients were treated with Gamma Knife radiosurgery (GKRS) for pituitary adenomas, and of these, 23 patients (1 male, 22 female) had microadenomas. Twenty-two of these patients were followed up and endocrinological tests were available for 15 of these 22. Thirteen of these 15 had prolactinomas, while the remaining 2 had acromegaly. The follow-up period was from 3 to 26 months (median 12 months). The mean age was 33.6 years (range 21 to 60 years). The mean maximum tumor dose was 35.7 Gy and the mean margin dose was 22 Gy. Serum prolactin (PRL) was normalized in three patients, decreased in eight and unchanged in two. The growth hormone (GH) secretion in the acromegalic patients has remained unchanged through the follow-up period. Thus, GKRS is a valuable adjuvant to transsphenoidal microsurgery for patients with pituitary microadenomas. PMID- 10681700 TI - Pituitary adenomas treated by microsurgery with or without Gamma Knife surgery: experience in 122 cases. AB - The clinical outcome of 122 patients with pituitary adenomas treated by microsurgery and/or Gamma Knife radiosurgery (GKRS) was analyzed to evaluate patient selection criteria and the role of GKRS. Sixty-six resections were performed in 59 patients. All tumors were macroadenomas, except for 5 ACTH producing adenomas. Twenty-four of the 31 hypersecreting adenomas showed normal serum hormone values after treatment. Postoperative complications were rhinorrhea, cranial nerve palsies, and a small thalamic infarct. GKRS was performed on 18 of the operated patients because of residual tumors, mostly in the cavernous sinus. Thirty-five of the 63 patients treated by GKRS were followed for more than 2 years. All adenomas except 2 were stable or had decreased in size. Eleven of 17 functioning adenomas showed normal serum hormone values after treatment. It is concluded that tumors that compress the optic pathway should be removed and that residual tumors in the cavernous sinus are good indications for radiosurgery. PMID- 10681701 TI - Cerebral infarction with ICA occlusion after Gamma Knife radiosurgery for pituitary adenoma: A case report. AB - Cranial irradiation may lead to accelerated atherosclerotic changes to small or medium sized arteries, but stroke associated with pituitary irradiation is not frequent. A patient treated with Gamma Knife radio-surgery (GKRS) for a pituitary adenoma suffered a cerebral infarction with internal carotid artery occlusion 4 years after radiosurgery. The patient was a 35-year-old male presenting with a visual disturbance. Endocrinological tests were normal. MRI revealed a 4.3 by 4.3 cm diameter invasive macroadenoma of the pituitary, projecting toward the suprasellar region and with cavernous sinus involvement with encasement of both internal carotid arteries (ICAs). GKRS was performed for residual tumor after a transcranial resection. The maximum dose was 40 Gy and the dose to the right carotid artery was below 20 Gy. The delayed hemiparesis was accompanied by a right capsular lacunar infarct shown on MRI. The images also showed a marked reduction in tumor size. Total, right ICA occlusion was confirmed by Doppler ultrasound. The patient had no history or signs of heart disease or metabolic disorder which could predispose to cerebrovascular PMID- 10681702 TI - Craniopharyngiomas: A six year experience with Gamma Knife radiosurgery. AB - The focused radiation produced by radiosurgery offers the theoretical advantage of a reduced radiation dose to surrounding structures during the treatment of residual craniopharyngiomas when compared with fractionated radiotherapy. A retrospective analysis of 23 patients treated since 1992 was undertaken. Intracystic bleomycin treatment in 10 patients with cystic tumors proved to be effective in preparing the tumors for radiosurgery. The mean prescription dose in this series was 10.8 ? 8.7 Gy delivered to a mean prescription 47% isodose. There was a volume reduction of the residual tumor in 74%. In this series, initial tumor volume and target volume were significant prognostic factors. Smaller tumors and targets were more likely to shrink. Five patients with large multicystic residual or recurrent tumors after multiple surgical attempts showed further progression. The maximum radiation dose, prescription dose and choice of prescription isodose were not significant prognostic factors within this series. Radiosurgery resulted in no mortality and no significant morbidity could be attributed to it. Clinically, the best results with radiosurgery were obtained with monocystic tumors amenable to stereotactic drainage and intracystic bleomycin treatment. PMID- 10681703 TI - TLD measurements of the relative output factors for the Leksell Gamma Knife. AB - The accuracy of the output factor directly affects the accuracy of dose delivery during patient treatment. At our Gamma Knife center, annual output factor measurements have been carried out by using a high accuracy TLD technique, characterized by the group annealing and sorting (GAS) procedure. For each collimator exposure, one to five LiF Thermoluminescent Dosimeter (TLD) 1 x 1 x 1 mm3 cubes were used, depending upon the collimator size, and the process was repeated until approximately 10 TLDs had been exposed for each collimator. Transit radiation dose accumulated during the motion of the treatment couch was measured for each collimator helmet, and the result was subtracted from the uncorrected TLD dose measurements. The mean values of the output factors for the 14, 8 and 4 mm collimators from last 5 years were 0.985 ? 0.001, 0.948 ? 0.005 and 0.833 ? 0.007, respectively, relative to the 18 mm collimator. These measured relative output factors are virtually identical to the values recommended by the manufacturer for the 14 mm and 8 mm collimators. On the other hand, the output factor for the 4 mm collimator was approximately 4.1% larger than that recommended by the manufacturer. The significance of these findings is discussed. PMID- 10681704 TI - Successful conversion from a linear accelerator-based program to a Gamma Knife radiosurgery program: the Cleveland Clinic experience. AB - From August 1989 to January 1997, 307 treatments in 293 patients were performed with a linear accelerator-based (LINAC) stereotactic radiosurgery system. Because of the program s success, the need for a dedicated radiosurgery unit in Ohio and the desire to treat functional disorders, the Cleveland Clinic Health System (CCHS) obtained the first Gamma Knife in the state of Ohio. Based on the previous volume of patients for radiosurgery, it was estimated that 75-100 patients would be treated during the first year of operation. However, during the first calendar year, 214 treatments were performed on 205 patients, which far exceeded expectations. The success of the CCHS Gamma Knife Center can be attributed to an increase in a number of factors. These included marketing efforts, patient awareness, increased use for functional disorders, physician understanding of radiosurgery, use by qualified nonaffiliated radiation oncologists and neurosurgeons, and outpatient delivery (95% with the Gamma Knife vs <5% with the LINAC). With proper planning, education, and awareness, the opening of a Gamma Knife Center can greatly increase the volume of radiosurgery performed when compared with a LINAC-based program. PMID- 10681705 TI - A case of very large cyst formation with Gamma Knife radiosurgery for an arteriovenous malformation. AB - A 17-year-old male patient underwent Gamma Knife radiosurgery (GKRS) for a left parietal arteriovenous malformation (AVM), which presented with hemorrhage. The 15.0 cm3 nidus was covered with the 50% isodose. The maximum dose was 50 Gy and the margin dose was 25 Gy. Eleven months later he developed a right hemiparesis and MRI showed a large cyst. Cerebral angiography showed partial obliteration of the AVM nidus. Stereotactic removal of cyst fluid (about 70 cm3) was performed, and an Ommaya reservoir was inserted. Cyst formation after GKRS for cerebral arteriovenous malformation is a is side effect of radiosurgery about which we need to learn more. PMID- 10681706 TI - The role of Gamma Knife radiosurgery in arteriovenous malformation with aneurysms. AB - A review of 217 patients treated with Gamma knife radiosurgery (GKRS), at Hospital Na Homolce, Prague, between October 1992 and January 1998 for arteriovenous malformation (AVM) is presented. Forty-one patients (18.9%) with an AVM and associated aneurysm are the subjects of special interest for this study. The nidus volume in the presence of an aneurysm lying close to the nidus or within it was significantly larger than the nidus volume in cases where the AVMs had no associated aneurysm, suggesting that an increased flow in a larger AVM may be an important factor for aneurysm formation. The association of an arterial aneurysm with an AVM significantly increased the chance of hemorrhage when compared to the group with AVM and no aneurysm. Ten patients out of 14, who had an aneurysm close to or within the nidus, showed a complete obliteration of their AVM and aneurysm, although the latter was not always included within the irradiated volume. Thus, this study indicates that radiosurgery alone could be the method of choice for the treatment of a combination of AVM and aneurysm, if the aneurysm is close to or within the nidus. PMID- 10681707 TI - [The advance of molecular technology in clinical microbiology laboratories]. PMID- 10681708 TI - The isolation results and the antimicrobial agents susceptibility analysis of Haemophilus from the sputum of the elderly suffering from respiratory tract disease. AB - OBJECTIVE: To inform the isolation and antibiotics susceptibility results of Haemophilus in the sputum of old men suffering from respiratory tract disease so that the epidemiological information and treatment can be available to physicians. METHODS: Two groups-group A, patients=562, group B, healthy volunteers=57 expectorated specimens of sputum were cultured with a special medium for Haemophilus. The isolates were then identified with API NH Identification plates and detected with ATB NH antibiotics susceptibility cards, finally, we got the statistics data of Haemophilus species and antibiotics susceptibility rates from Datatrac statistic system of VITEK 32. RESULTS: Haemophilus were detected in 132 specimens out of 562 (23.5%), which includes 6 strains of H. influenzae (4.5%). 122 strains of H. parainfluenzae, 4 strains of H. aphrophilus (3.1%). At the same time, 3 strains of Haemophilus were isolated from group B. Following are the susceptibility rates of sixteen antibiotics of 132 strains of Haemophilus, 73.4% to amoxicillin, 92.4% to amoxicillin and clavulanic acid, 83.3% to cefactor, 90.9% to cefotaxime, 79.5% to cefuroxime axetil, 52.3% to kanamycin, 25.0% to gentamycin, 72.0% to spectinomycin, 79.5% to chloramphenicol, 50.0% to tetracycline, 9.1% to erythromycin, 42.4% to pristhomycin, 49.2% to rosoxacin, 62.96% to pefloxacin, 97.7% to rifampin, 33.3% to sulfonamides. CONCLUSION: There is a high isolation rate of Haemophilus in the sputum of the elderly suffering from respiratory tract diseases, The H. parainfluenzae is the main Haemophilus in our area of study, which it deserves good attention by clinical microbiological laboratory. In addition, the antibiotics susceptibility results indicate that this species is more susceptible to beta-lactams antibiotics and is less susceptible to some others such as macrolides and aminoglycosides, etc. So, the drug treatment should be based on these antibiotics susceptibility results in order to guide the specific therapy. PMID- 10681709 TI - Identification of Lactobacillus species of human origin by a commercial kit, API50CHL. AB - The efficacy of a biochemical kit, API50CHL kit, for identification of intestinal and vaginal lactobacilli from humans was evaluated by comparing with the results of DNA-DNA hybridization assay. The results showed that in total only 52 of the 172 strains (30.2%) tested were identified correctly by the kit at species level. Especially all strains of some species, such as Lactobacillus gasseri and Lactobacillus crispatus, were misidentified as Lactobacillus acidophilus by the kit. However, the kit was found to be reliable for identification of Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius. This suggests that the exact identification of Lactobacillus isolates from human stool and vaginal specimens by API50CHL kit is difficult without the support of modern genotypic technique. PMID- 10681710 TI - [Evaluation of inoculum density prepared by prompt inoculation system and antimicrobial susceptibility test results by the automated MicroScan WalkAway system]. AB - The Prompt Inoculation System adapted to the susceptibility testing by the automated microbiology system, MicroScan WalkAway (Dade MicroScan Inc., West Sacramento, CA, U.S.A.) was evaluated by determining colony forming units (cfu) per ml of the inocula and by the susceptibility test results obtained through repeated testing of the American Type Culture Collection (ATCC) reference strains described by the National Committee for Clinical Laboratory Standards (NCCLS). The colony forming units per ml of the inocula prepared by the Prompt ranged 2x10++(5) to 2x10(6)++ cfu/ml for the ATCC reference strains, the results indicating that the Prompt gave a higher inoculum density and was more reproducible when compared to the standard turbidity, McFarland adjustment. Also, most inocula prepared from the clinical isolates, comprising the strains of Enterobacteriaceae,no-entericbacilli,staphylococci,enterococci, and streptococci,contained 1x10(6) to 3x10(6) cfu/ml. Although the inocula prepared by the Prompt contained more viable bacterial cells, the outcome results for susceptibility testing by the MicroScan WalkAway were highly acceptable. Four ATCC reference strains were repeatedly tested. Of 540 MIC determinations, 489 (90. 6%) were within the acceptable MIC ranges described by the NCCLS M100-S9, whereas the inocula prepared by the photometric adjustment gave 87.4%. In conclusion, the Prompt inocula were found to give more precise susceptibility test results mostly equivalent to those obtained from inocula prepared by the conventional photometric procedures. PMID- 10681711 TI - [Rapid detection of Mycoplasma pneumoniae-specific IgM]. AB - The rapid diagnosis of Mycoplasma pneumoniae infection is important in carrying out chemotherapy in appropriate manner. It is also essential to detect the specific immunoglobulin M (IgM) in order to diagnose infectious diseases. The ImmunoCard Mycoplasma kit (TFB. Inc./Meridian Diagnostics, Inc.) is a 10-min-card based enzyme-linked immunosorbent assay (ELISA) of IgM antibodies to M. pneumoniae. The ImmunoCard was compared with high density particle agglutination (HDPA) and cold hemagglutinin (CHA). The ImmunoCard test had 98.3% sensitivity, 51.4% specificity, and 72.5% agreement with HDPA (>or =320), but it had 94.3% sensitivity, 87.5% specificity, and 92.2% agreement with clinical diagnosis. Our results indicate that the ImmunoCard Mycoplasma IgM assay is a rapid, simple and valuable procedure which can be analyze small numbers of specimens using a less complicated technique and with no other equipment required. This means that the ImmunoCard is a cost-effective, energy saving and rapid procedure for the detection of M. pneumoniae-specific IgM. PMID- 10681712 TI - [Evaluation of direct identification testing from positive blood culture bottles with MicroScan rapid identification panels]. AB - We evaluated the direct identification method from growth-positive blood culture bottles using MicroScan Rapid ID panels (DADE BEHRING) for the purpose of rapid identification. The inoculum for Rapid ID panels were prepared using an isolation method from blood culture bottles by VACUTAINER (BD). McF 1.0 had a better result than McF 0.5 as the inoculum concentration for Rapid ID panels. Rapid ID panel identification results were effected by blood contamination for > or =0.3% of S. aureus and 0.9% of a strain of E. coli. Blood contamination from the bottle may cause an issue to the identification results. The accuracy of this direct identification testing was 72.0% (36 out of 50) for gram positives organisms and 88. 9% (80 out of 90) for gram negatives organisms. Although some strains including S. pyogenes, coagulase-negative staphylococci and non-Fermentative Gram Negative Rods had not identified correctly, this method provides a preliminary result within 3 hours and provides a fast turn around time. In conclusion, this method was considered as an effective method for routine testing. PMID- 10681713 TI - [Evaluation of a fully automated mycobacteria culture system, MB/BacT using a newly developed digestion-decontamination procedure, semi-alkaline protease-N acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) method]. AB - A fully automated non-radiometric mycobacteria culture system, MB/BacT (Organon Teknika, Durham, NC, U.S.A.), was recently introduced in Japan and evaluated for its ability to detect mycobacteria in clinical sputum specimens. A previous study yielded nearly a 40% contamination ratio from sputa treated with the standard N acetyl-L-cysteine (NALC)-NaOH method. This study employed a mucolytic agent (semi alkaline protease; SAP) in which the sputa were processed twice for digestion followed by decontamination at twice the standard volume of NALC-NaOH. The concentrated sediments were resuspended in phosphate buffer (0.067 M, pH 6.8), and inoculated into the MB/BacT Process Bottles supplemented with antibiotics. The bottles were incubated at 37 degrees C and monitored for up to fifty-six days. Recovery of mycobacteria was compared in three different egg-based Ogawa media in addition to a non-selective Middlebrook 7H10 agar. A total of 1, 124 clinical sputum specimens have been evaluated. Of these, 464 were positive for growth of mycobacteria, of which 447 (96.3%) were positive by the MB/BacT. False positive alarms due to break through contamination, mainly by Pseudomonas aeruginosa and Candida spp., were observed in twenty-one specimens (1.9%). The three Ogawa media could detect only 283 (60.5%) to 353 (75.4%) positives, and Middlebrook 7H10 agar only 424 (90.6%) positives. The time to detect positive cultures of Mycobacterium tuberculosis complex by the MB/BacT ranged from 2.2 days to 52.3 days, and 50% of positive cultures were detected within 16.7 days of incubation. It can be concluded that the combination of SAP-NALC-NaOH digestion decontamination procedure and the MB/BacT is particularly useful for the isolation of mycobacteria and has a faster time to detect than conventional methods. MB/BacT is a suitable alternative method for the detection of mycobacteria in Japan, where the radiometric Bactec System is not available. PMID- 10681714 TI - [Detection of pyrazinamidase activity for differentiation of Campylobacter, Arcobacter, and Helicobacter spp. by using a high-performance liquid chromatography method]. AB - A high-performance liquid chromatography method was investigated for the detection pyrazinamidase activity by Campylobacter, Arcobacter, and Helicobacter spp. Pyrazine carboxilic acid, one of the end products of pyrazinamide hydrolysis by microorganisms, was detected by using a high-performance liquid chromatography (HPLC). A loopful of organism colonies was emulsified in 0.5 ml of a 0.5% pyrazinamide solution. The suspens on was incubated in a 37 degrees C water bath for 18-20 hr. After centrifugation, the supernatant was analyzed by HPLC. This HPLC method does not require microaerobic incubation and was easy to interpret for strains with weak enzymatic activity. By this method, we tested 111 clinical isolates, type and reference strains of Campylobacter spp., Arcobacter spp., and Helicobacter spp. , C. jejuni, C. jejuni subsp. doylei, C. coli, C. upsaliensis, C. lari, C. lari (urease+), C. helveticus, C. hyolei, C. sputorum subsp. fecalis, C. gracilis, C. concisus, C. curvus were positive for pyrazinamidase. C. fetus subsp. fetus, C. hyointestinalis, C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, C. mucosalis, A. butzleri, A. skirrowii, A. cryaerophilus, H. pylori, H. cinaedi, H. fennelliae, H. mustelae, H. felis, H. muridarum, H. canis, H. nemestrinae, H. pamentensis, H. pullourum were negative. PMID- 10681715 TI - Indolent lymphomas. PMID- 10681716 TI - Chelation therapy in patients with thalassemia using the orally active iron chelator deferiprone (L1). AB - BACKGROUND AND OBJECTIVE: Excessive hemosiderosis is the main reason for the multi-organ failure observed in multitransfused patients. Deferiprone (1,2 dimethyl-3-hydroxy-pyridine-4-one, L1) is an orally active iron chelator mainly excreted via urine. We conducted a study in order to determine the efficacy and safety of L1 in Greek thalassemic patients. DESIGN AND METHODS: A group of 11 thalassaemic patients entered the study; L1, the Cipla formulation for deferiprone, at a daily dose of 75-100 mg/kg bw t.i.d. was used. After giving informed consent all patients were subjected to clinical examination and biological tests. RESULTS: All patients tolerated the L1 well; there were no significant side effects (except for slight gastrointestinal disturbances for the first days). The net urinary iron excretion ranged from 6.96 to 26.1 mg/24h. Serum ferritin declined within 4-6 months in most of the patients. INTERPRETATION AND CONCLUSIONS: The results suggest that L1 is a rather safe drug which decreases iron overload without causing any considerable side-effects in Greek thalassemics. PMID- 10681717 TI - Analysis of T-cell clones in systemic lupus erythematosus. AB - BACKGROUND AND OBJECTIVE: It is fairly well established that T-helper (TH)((1)) cells play a role in the pathogenesis of organ-specific autoimmune diseases, while their role and their relationship with TH((2)) cells is far from being defined in systemic lupus erythematosus (SLE). To address this issue, six female patients who fulfilled the American Rheumatism Association criteria for the diagnosis of SLE were studied. DESIGN AND METHODS: We analyzed the intracellular production of cytokines by T-cells from the peripheral blood (PB). Then, we established T-cell clones (TCC) from the peripheral blood (PB) of all cases as well as from the synovial fluid of one patient with an articular flare-up. RESULTS: The percentages of IL-4 positive and IFN-g positive PB T-cells were not different between SLE patients and normal controls. When 93 TCC (67 CD4(+), 23 CD8(+)) from the PB of 5 different SLE patients were compared to 118 TCC (94 CD4(+), 23 CD8(+)) from 5 healthy controls no statistical difference was observed between SLE and controls in terms of TH((1)), TH((2)) or TH((0)) phenotype. However, SLE clones showed a reduced ability to secrete IL-10 (p = 0. 002). In contrast, the analysis of the 30 clones obtained from synovial fluid revealed that 11/23 CD4(+) clones were TH((1)), 12/23 were TH((0)), 2/7 CD8(+ )clones were TH((1)) and 5/7 were TH((0)). No TH((2)) clones were obtained from the synovial fluid. INTERPRETATION AND CONCLUSIONS: The data suggest that the T-cell subsets operating in actively inflamed organs of SLE may belong to the TH((1)) and TH((0)) subsets. PMID- 10681718 TI - Heterogeneity of isolated mononuclear cells from patients with acute myeloid leukemia affects cellular accumulation and efflux of daunorubicin. AB - BACKGROUND AND OBJECTIVE: Pharmacologic studies on blasts from patients with leukemia are generally performed on density gradient isolated blood or bone marrow cells. Thereby, cellular drug accumulation and efflux are determined as mean values of the entire cell population. The objective of the present study was to characterize the heterogeneity in the accumulation and efflux of daunorubicin in various subpopulations of mononuclear cells isolated from patients with acute myeloid leukemia (AML). DESIGN AND METHODS: Mononuclear cells from 33 patients with AML were isolated from peripheral blood by density gradient centrifugation on Lymphoprep (1. 077 g/mL). Cellular accumulation of fluorescent daunorubicin was determined by flow cytometry after incubation of the cells at +37C for 1 hour. Thereafter, the cells were washed and reincubated in drug-free medium. Kinetics of drug efflux were determined by frequent determination of cellular fluorescence during 30 min. Daunorubicin accumulation and efflux were compared in the total isolated mononuclear cell population and in the various blast cell populations gated on FSC/SSC according to the results of immunophenotyping. RESULTS: In 8 of these 33 (24%) patient samples, two distinct blast cell populations could be identified. In 7 out of 8 these cases the more immature blasts had a lower drug accumulation and in 6 out of the 8 cases also a higher efflux rate than the differentiating cell population. Cyclosporin A increased daunorubicin accumulation and reduced efflux in the immature blast population. In the differentiating cell population cyclosporin A increased both the accumulation and the efflux. In patients with a single blast cell population, the gated blast cells had a significantly lower drug accumulation but also a lower drug efflux rate than the total cell population. INTERPRETATION AND CONCLUSIONS: The results imply that drug transport studies on cells isolated from patients with AML give somewhat different results depending on the cell population studied. Some, but not all, of these differences in daunorubicin accumulation and efflux as well as in the effect of cyclo-sporin A can be explained by a heterogenous expression of the mdr1-gene. The observed heterogeneity may be of special relevance with regard to drug resistance. The presence of even a small resistant cell clone may jeopardize the effect of the chemotherapy due to expansion resulting in relapse of disease. PMID- 10681719 TI - Prolonged response to cyclosporin-A in hypoplastic refractory anemia and correlation with in vitro studies. AB - BACKGROUND AND OBJECTIVE: Lymphocyte abnormalities in myelodysplastic syndromes (MDS) have been widely described, but the role of the immune system in the pathogenesis of these clonal disorders remains controversial. An active role of lymphocytes in suppressing normal hematopoiesis may be implicated in MDS with hypoplastic marrow. We have studied in vitro and in vivo activity of cyclosporin A (CSA) on hematopoiesis in patients affected by hypoplastic MDS without blast excess. DESIGN AND METHODS: Nine consecutive patients with hypoplastic refractory anemia (RA), followed up in our out-patient unit, were treated with CSA at daily doses of 1-3 mg/kg for at least three months. Low dose steroids or danazol were transiently added in 7/9 patients. Differences between pre- and post-treatment parameters were studied by the Student's t-test. In vitro effect of CSA on circulating hematopoietic progenitors was studied by the methylcellulose colony assay. RESULTS: Before treatment, fewer circulating hematopoietic progenitors were found in all patients as compared to normal subjects. The number of CD34+ cells was about halved, while circulating erythroid and myeloid colony-forming cells (CFC) were reduced to one-fifth. After a mean period of 22 months of CSA treatment (median: 14.5 months), hemoglobin was significantly and persistently increased in two patients, platelets in one, platelets and hemoglobin in two. Two patients showed transient responses, one patient did not tolerate the treatment and one patient is close to a significant response. At in vitro CSA concentrations similar to those achieved in vivo after oral administration the drug significantly increased cell colony growth in hypoplastic RA. This test correctly predicted a positive clinical response to CSA in 3/5 cases and treatment failure in 4/4 cases. INTERPRETATION AND CONCLUSIONS: About one half of hypoplastic RA patients benefited from CSA treatment. A larger study could verify whether in vitro culture of hematopoietic progenitors in the presence of CSA can predict the clinical response and whether this treatment could prolong patients' survival. PMID- 10681720 TI - L-selectin expression is low on CD34+ cells from patients with chronic myeloid leukemia and interferon-a up-regulates this expression. AB - BACKGROUND AND OBJECTIVE: Altered adhesive interaction between bone marrow (BM) stroma and progenitors in chronic myeloid leukemia (CML) may be in part caused by abnormal expression of cell adhesion molecules (CAMs) on malignant progenitor cells. Treatment of CML with interferon-a (IFN-a) re-establishes normal hemopoiesis in some patients in part by restoring normal adhesive interactions between CML progenitors and BM microenvironment, which may in turn be mediated by correcting CAM expression on progenitors. DESIGN AND METHODS: We investigated the expression of CAMs (L-selectin, b((2))-integrin, LFA-3, ICAM-1, ICAM-3, NCAM) on purified BM CD34(+) cells from CML patients (n= 34) and healthy adults (n= 15) by flow cytometry. Modulation of L-selectin expression on CD34(+) cells from CML after in vitro treatment with IFN-a was also investigated. RESULTS: The mean percentage of CD34(+ )cells expressing L-selectin was significantly lower in CML patients (25.4+/-12.8%) than in normal controls (68.7+/-8.3%, n=15). CD34(+)/HLA DR(&endash;/low) and CD34(+)/ CD38(&endash;/low) co-expressing L-selectin were also significantly lower in untreated CML (27.4+/-21.5% and 39.8+/-26.7%, respectively, n=8) than in controls (61+/-17% and 83.7+/-10%, respectively, n=7). In vitro treatment with IFN-a of purified CD34(+) BM cells from untreated CML patients (n=8) induced a significant, dose and time-dependent increase in the L selectin expression as indicated by FACS analysis. INTERPRETATION AND CONCLUSIONS: We hypothesize that this L-selectin deficiency reflects a cell surface adhesion defect of progenitors from CML that is partially restored by in vitro IFN-a treatment. These data may help to explain the adhesive abnormalities of CML progenitors to the BM microenvironment and the in vitro restoration of adhesion capacity after IFN-a treatment. PMID- 10681721 TI - Prevalence of Helicobacter pylori and hepatitis C virus infections among non Hodgkin's lymphoma patients in Southern Switzerland. AB - BACKGROUND AND OBJECTIVE: Several recent studies have reported a high rate of previous hepatitis C virus (HCV) infection in patients with non-Hodgkin's lymphoma (NHL). However, it appears that there are marked geographical differences in the prevalence of HCV among NHL patients. There is further controversy concerning a possible pathogenetic link between HCV and certain histologic lymphoma subtypes, in particular MALT lymphomas, and it has recently been speculated that HCV might be involved in the multistep process of gastric lymphoma genesis, in addition to the well established role of chronic Helicobacter pylori infection. The aim of this study was to investigate the prevalence of HCV and H. pylori infections in patients with B-cell NHL in Southern Switzerland. DESIGN AND METHODS: One hundred and eighty newly diagnosed HIV-negative B-cell NHL patients, consecutively seen at a referral oncology center in Southern Switzerland between 1990 and 1995 were prospectively studied. A microparticle enzyme immunoassay was used to detect antibodies to HCV. Serologic determination of HCV genotype was done by the Murex method. The quantitative detection of IgG anti-H. pylori was performed by the Biorad GAP test. RESULTS: Infection with HCV was detected in 17/180 patients (9.4%; 95% C.I., 6%-15%). This prevalence is significantly higher than that observed in a large survey of 5424 new blood donors from the same area tested in 1992-97 (0.9%; 95% C.I., 0.7-1.2). Neither histologic subtypes nor specific extranodal presentations of NHL were associated with a higher prevalence of HCV. HCV serotype 2 (corresponding to genotypes 2a-c) was the most common. HCV infection was significantly associated with a shorter progression-free survival at both univariate and multivariate analysis. Anti-Helicobacter antibodies were detected in 81/180 patients (45%; 95% C.I., 38%-53%) and H. pylori infection was significantly associated with the development of primary lymphomas of the stomach. INTERPRETATION AND CONCLUSIONS: A high prevalence of HCV infection was detected in NHL lymphoma patients and was associated with a shorter time to lymphoma progression. HCV infection was not correlated with primary gastric presentation or with MALT-type histology. Our findings further support the key role of H.pylori infection in the pathogenesis of primary gastric lymphoma of MALT-type. The possible role of HCV in the pathogenesis of NHL should be further investigated. PMID- 10681722 TI - Validation of the international prognostic index in working formulation group A low-grade non-Hodgkin's lymphoma: retrospective analysis of 137 patients from the Gruppo Italiano per lo Studio dei Linfomi registry. AB - BACKGROUND AND OBJECTIVE: The subset of non-follicular non-Hodgkin's lymphoma (NHL) includes patients with varied prognoses, thus suitable for different therapeutic approaches. The International Prognostic Index (IPI), originally proposed for aggressive NHL, has been demonstrated to be of prognostic relevance also in follicular NHL. The main aim of the study was to validate the IPI in this histologic category; in addition, the specific prognostic classification, currently employed in the Gruppo Italiano per lo Studio dei Linfomi (GISL) prospective therapeutic trials and based on different features, more similar to those applied to chronic lymphocytic leukemia, was analyzed. DESIGN AND METHODS: The present series consists of 137 evaluable patients affected by Working Formulation group A NHL out of 256 cases referred to the GISL Registry. The retrospective prognostic study included the evaluation by both univariate and multivariate analyses of overall survival, response to therapy and response duration. The IPI was applied as originally proposed. The GISL definition of indolent and aggressive disease at diagnosis was based on the presence of B symptoms, bulky disease, anemia and thrombocytopenia. RESULTS: The distribution of patients in IPI risk groups was rather unbalanced with 18%, 47%, 28% and 7% of cases classified as low (L), intermediate-low (IL), intermediate-high (IH) and high (H) risk, respectively. The median overall survival was not reached in either L or IL risk groups, and was 84.1 and 7.4 months for IH and H risk groups, respectively (p=0. 0005). A simplified IPI model was designed merging patients in both intermediate risk groups and the statistical difference of survival retained its significance. GISL prognostic stratification was demonstrated to have a significant association with survival, with a median survival of 71.3 months in aggressive disease and a median survival not reached at 152 months in indolent disease. Both the simplified IPI model and the GISL risk definition retained their significance in multivariate analysis for overall survival, while for response to therapy only the simplified IPI model resulted to be of statistical significance. In addition, the GISL prognostic stratification identified patients with different outcomes within the IPI intermediate risk group, with a median survival of 70.2 months for patients with aggressive disease wheras the median survival for those with indolent disease was not reached. Finally, a prognostic score resulting from the integration of the simplified IPI and the GISL system was statistically validated. INTERPRETATION AND CONCLUSIONS: The retrospective analysis of this series demonstrates the validity of the IPI in non-follicular indolent NHL and the usefulness of integrating the IPI parameters with disease specific prognostic variables. PMID- 10681723 TI - VACOP-B, high-dose cyclophosphamide and high-dose therapy with peripheral blood progenitor cell rescue for aggressive non-Hodgkin's lymphoma with bone marrow involvement: a study by the non-Hodgkin's Lymphoma Co-operative Study Group. AB - BACKGROUND AND OBJECTIVE: Sequential treatment with the addition of high-dose therapy (HDT) and peripheral blood progenitor cell (PBPC) rescue has been reported to be active as front-line therapy in aggressive non-Hodgkin's lymphoma (NHL) with bone marrow (BM) involvement. We designed an intensive sequential therapy as front-line therapy in this subset of patients and conducted a phase II study. DESIGN AND METHODS: Patients with aggressive non-Hodgkin's lymphoma and BM involvement at diagnosis received 8 weeks of VACOP-B chemotherapy as induction therapy. The second phase included high-dose cyclophosphamide (HDCY) (7 g/m(2)) with granulocyte colony-stimulating factor (G-CSF) followed by leukaphereses. The third phase included HDT according to the BEAM protocol or melphalan (140 mg/m(2)) plus total body irradiation (8 Gy in a single dose). RESULTS: Forty patients were included in the study. According to the intention-to-treat, after VACOP-B, 11 (27.5%) and 22 (55%) patients achieved complete remission (CR) and partial remission (PR), respectively. Thirty-four received HDCY. After HDCY, 18 patients (45%) were in CR and 13 (32.5%) in PR. Twenty-nine underwent HDT plus peripheral blood cell rescue (PBPC) rescue. At the completion of treatment 29 patients (72.5%) were in CR, and 3 patients (7.5%) in PR. The actuarial 3-year overall survival, disease free survival and failure free survival are 48%, 55% and 40%, respectively. Overall severe toxicity was 7.5%. INTERPRETATION AND CONCLUSIONS: This phase II study suggests that the intensified treatment described is feasible and active in aggressive NHL with BM involvement. A randomized trial is now underway to test this approach. PMID- 10681724 TI - Autologous stem cell transplantation for high-risk Hodgkin's disease: improvement over time and impact of conditioning regimen. AB - BACKGROUND AND OBJECTIVE: High-dose chemo/radiotherapy with autologous stem cell support is increasingly being used in Hodgkin's disease (HD) patients who do not respond to or who relapse after conventional chemotherapy. In this work we analyze the results of 56 consecutive high-risk HD patients autografted in our institution and the role of possible prognostic factors. DESIGN AND METHODS: There were 34 males and 22 females with a median age of 31 years. At transplantation, 24 patients (43%) were in complete remission and 32 (57%) were autografted while with active disease. Twenty-nine patients were autografted before January 1993. Bone marrow was used as the source of stem cells in 40 patients (71%) and peripheral blood (PB) in 16 (29%). Forty-five patients received chemotherapy-based conditioning regimens (40 CBV and 5 BEAM) while the remaining 11 received cyclophosphamide (Cy) and total body irradiation (TBI). RESULTS: Two bone marrow transplantation (BMT) recipients did not engraft. Hematologic recovery was significantly faster in patients transplanted with PB progenitor cells. Early transplant-related mortality (early TRM) (before day 100 after transplantation) was 9%; it was higher in patients transplanted before January 1993 than in patients transplanted afterwards (14% vs 4%) and in patients receiving TBI (18% vs 7%), although these differences did not reach statistical significance. Overall TRM (before and after day 100) was 14%. TBI-containing regimens significantly increased overall TRM (36% and 9%, p = 0.03). Actuarial 3.5-year overall survival (OS), event-free survival (EFS) and progression-free survival (PFS) were 57%, 58% and 65%, respectively. On multivariable analysis, TBI containing regimens and transplantation before 1993 significantly reduced OS and EFS. INTERPRETATION AND CONCLUSIONS: Our results confirm that high-dose therapy followed by autologous stem cell transplantation is associated with sustained PFS in a remarkable proportion of patients with HD unlikely to be cured with standard chemotherapy. Results improved over time and TBI containing regimens had a negative effect on post-transplant outcome. PMID- 10681725 TI - Myeloid mixed chimerism is associated with relapse in bcr-abl positive patients after unmanipulated allogeneic bone marrow transplantation for chronic myelogenous leukemia. AB - BACKGROUND AND OBJECTIVE: Although bcr-abl polymerase chain reaction (PCR) positivity after bone marrow transplantation (BMT) for chronic myelogenous leukemia (CML) is significantly related to relapse, the predictive value of the assay is not very high and therefore most investigators consider that qualitative RT-PCR data alone are too imprecise to enable clinical decisions to be taken in individual cases. To define the clinical outcome of bcr-abl positive patients after unmanipulated BMT better, we sought the origin of hematopoiesis and traced its evolution over time. DESIGN AND METHODS: Forty-nine patients received allogeneic BMT for CML (39 in chronic phase and 10 in accelerated phase/blast crisis). Median follow-up was 61 months (range 4-92). mRNA and DNA were used to assess bcr-abl and chimerism status respectively. Quantitative VNTR-PCR on total cells and lymphoid or myeloid population allowed us to assign and measure the origin of hematopoiesis. RESULTS: Both bcr-abl positivity and the presence of mixed chimerism (MC) were significantly associated with relapse (p = 0.0009 and p < 0.0001 respectively). Relapse was observed in one of 39 patients with complete donor chimerism and in 6 of 9 patients with MC. These six cases showed increasing levels of host hemopoiesis and bcr-abl positivity in the CD15-positive population prior to relapse. The other three cases had decreasing or stable low-level MC which was restricted to the T-cells as well as bcr-abl negativity. INTERPRETATION AND CONCLUSIONS: Whereas the simple detection of bcr-abl fails to identify patients who will relapse with certainty, the assessment of MC by VNTR-PCR does identify patients headed to relapse. Confirmation of myeloid involvement and increasing levels over time further elucidates the clinical outcome of bcr-abl positive patients after BMT. PMID- 10681726 TI - Prevalence of TT viral DNA in italian blood donors with and without elevated serum ALT levels: molecular characterization of viral DNA isolates. AB - BACKGROUND AND OBJECTIVE: A novel non-enveloped DNA virus, called TT virus (TTV), has been reported to be associated with post-transfusion hepatitis of unknown etiology. Although its clinical role still remains obscure, its presence in blood donations might cause problems. It, therefore, appeared of interest to investigate TTV prevalence in voluntary blood donors. DESIGN AND METHODS: A total of 595 Italian blood donors with and without elevated serum alanine aminotransferase (ALT) levels were tested by polymerase chain reaction using two sets of semi-nested primers that amplify the well-known region in the N22 clone. The amplified products were then sequenced to assess the genotype by phylogenetic and restriction fragment length polymorphism analyses. RESULTS: The prevalence of TTV in blood donors was 5+/-1.9% (25 out of 500) with a 95% confidence limit. A similar prevalence was found in 95 selected blood donors with increased ALT levels. A viral load of 10(3)-10(4) viral DNA molecules/mL was found, thus indicating a rather narrow range of variability. A phylogenetic tree built up on the basis of 210 base sequences of ORF1 allowed isolates to be classified into 2 groups corresponding, at least, to two of the putatives TTV genotypes, group 1 and group 2 of Okamoto's classification. A similar classification was also obtained by site restriction enzyme analysis. INTERPRETATION AND CONCLUSIONS: The results show that TTV infection is present among Italian blood donors. No significant difference in prevalence of TTV infection was found between patients with normal and increased ALT, making the association between TTV infection and human hepatitis questionable. PMID- 10681727 TI - Relative sensitivity of direct antiglobulin test, antibody's elution and flow cytometry in the serologic diagnosis of immune hemolytic transfusion reactions. AB - BACKGROUND AND OBJECTIVE: Current immunohematology practice dictates that serologic diagnosis of immune hemolytic transfusion reactions (IHTR) is based on the finding of a positive post-transfusion direct antiglobulin test (DAT). However, since DAT may fail to detect antibody-coated cells when they constitute a minor population amid a large number of non-sensitized ones, we investigated whether antibody detection in eluates or by flow cytometry is more sensitive than DAT in this context. DESIGN AND METHODS: Ten samples of red blood cells sensitized with allo- or autoantibodies were diluted in non-sensitized red blood cells to final concentrations ranging from 10% to 0.1%. DAT, antibody detection in eluates, and immunofluorescence by flow cytometry were performed on each mixture. RESULTS: DAT failed to detect sensitized cells in all but two cases in that only the 10% dilution yielded a positive DAT. Antibody detection in eluates and by flow cytometry was able to detect up to 1% sensitized cells in most cases. INTERPRETATION AND CONCLUSIONS: Antibody detection in eluates and by flow cytometry is more sensitive than DAT for detecting minor populations of IgG coated cells. These techniques should be included in the routine investigation of suspected cases of IHTR. PMID- 10681728 TI - Alternative therapies and the Di Bella affair in pediatrics. A questionnaire submitted to Italian pediatric oncologists and hematologists. AB - BACKGROUND AND OBJECTIVE: Over the last 2-3 years in particular, the so-called Di Bella therapy (DBT) become the most famous of alternative treatments applied to pediatric oncology and hematology in Italy. Many Italian oncologists and hematologists had to cope with the problems that it introduced and the treatment also elicited heated reactions all over Europe. We attempted to evaluate the impact of this treatment on children with cancer. DESIGN AND METHODS: A questionnaire prepared with the aim of addressing the use of alternative therapies in pediatric hematology and oncology was circulated to the 48 centers (or divisions) belonging to AIEOP (Associazione Italiana di Oncoematologia Pediatrica) [Italian Pediatric Oncology and Hematology Association] and FONOP (Forza Operativa Nazionale di Oncologia Pediatrica) [National Pediatric Oncology Task Force]. The questionnaire consisted of 9 questions elaborated to give credit to the case-related and professional experiences of the colleagues we contacted. RESULTS: Forty-three centers replied to the questionnaire. Request to switch to DBT represented a considerable problem, involving the vast majority of centers participating into this study; however, case quantification varied greatly from center to center. One of the most significant aspects is that children switched to DBT, abandoning conventional therapies, were often relapsing or had had multiple relapses (from solid tumor or leukemia), but some children abandoned conventional therapies at an early stage and/or without fully exploiting the curative potential of these therapies. INTERPRETATION AND CONCLUSIONS: This study allowed us to obtain an evaluation of the impact of DBT in children with oncologic or hematologic disorders. It also highlights the importance of cultivating physician-parent dialogue and provides an opportunity for a few pedagogic thoughts on the attitude and opinions of pediatricians on this problem. PMID- 10681729 TI - Molecular pathophysiology of indolent lymphoma. AB - Indolent lymphomas are a markedly heterogeneous group of lymphoproliferative disorders including B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma, lymphoplasmacytoid lymphoma, follicular lymphoma, mantle cell lymphoma and mucosa-associated lymphoid tissue (MALT) lymphoma. The molecular pathophysiology of indolent lymphoma is characterized by distinct genetic pathways which selectively associate with different clinico-pathologic categories of the disease. At diagnosis, B-cell chronic lymphocytic leukemia frequently display deletions of 13q14, trisomy 12 and alterations of the ATM gene, whereas evolution to Richter's syndrome is associated with disruption of p53. Lymphoplasmacytoid lymphoma carries t(9;14) (p13;q32) in approximately 50% of cases, leading to the deregulated expression of the PAX-5 gene. Follicular lymphoma consistently harbors rearrangement of BCL-2. With time, a fraction of follicular lymphoma accumulates mutations of p53 and of p16 and evolves into a high grade lymphoma. MALT-lymphoma frequently associates with alterations of API2/MLT and, in some cases, of p53, BCL-6 and BCL-10. Studies of genotypic and phenotypic markers of histogenesis have shown that mantle cell lymphoma and a fraction of B-CLL/SLL derive from naive B-cells, whereas follicular lymphoma, lymphoplasmacytoid lymphoma and MALT-lymphoma originate from germinal center (GC) or post-GC B-cells. The identification of distinct genetic categories of indolent lymphoma may help in the therapeutic stratification of these disorders. In addition, genetic lesions of indolent lymphoma provide useful molecular markers for disease monitoring by high sensitivity techniques. PMID- 10681730 TI - Dendritic cells: specialized antigen presenting cells. AB - Renewing interest in cancer immunotherapy reflects the excellent results that have been obtained in animal models and the promising results in early clinical trails with dendritic cell (DC) based approaches. The central role that DCs play in the initiation of an immune response raises the possibility of using them to trigger specific anti-tumor immunity. In addition, deeper knowledge of DC biology will allow better understanding of the mechanism(s) underlying allergic and autoimmune diseases as well as tolerance phenomena. These crucial issues were critically reviewed during a workshop organized by the Italian Society for Experimental Hematology in Florence, Italy, on March 18th, 1999. The chairmen have prepared this report for the readers of Haematologica. PMID- 10681731 TI - Reversible adult respiratory distress in primary antiphospholipid syndrome. AB - Antiphospholipid antibody syndrome (APS) is a disorder caused by circulating antibodies reacting with biological membranes and characterized by recurrent thrombosis, chronic thrombocytopenia and miscarriages. It has been reported to occur either as a primary syndrome or secondary to systemic autoimmune disorders. We describe a case of primary APS in a young patient, in whom the clinical course was particularly severe and complicated by a respiratory distress syndrome. The patient was resistant to a number of treatments, and eventually responded to intravenous high dose corticosteroids. PMID- 10681732 TI - Unexpected consequence of splenectomy in composite lymphoma. The abscopal effect. PMID- 10681733 TI - Fatal bone marrow aplasia during interferon-a treatment in chronic myelogenous leukemia. PMID- 10681734 TI - Does interferon-a exert an anti-leukemic effect by enhancing cell mediated immunity in chronic myeloid leukemia? PMID- 10681735 TI - Absence of bcr/abl rearrangement in 41 patients with essential thrombocythemia. PMID- 10681736 TI - Cladribine as monotherapy or combined with dexamethasone and idarubicin or mitoxantrone in previously treated patients with low-grade lymphoid malignancies. PMID- 10681737 TI - Cyclophosphamide/cyclosporin-A treatment of multicentric Castleman's disease with Kaposi's sarcoma. PMID- 10681738 TI - High-dose ifosfamide and etoposide infusion plus methylprednisolone for refractory or relapsed aggressive non-Hodgkin's lymphoma. PMID- 10681739 TI - Transfusion requirement can be abolished by epoietin-a and autologous platelet predeposit in patients receiving high dose chemotherapy with stem cell support. PMID- 10681740 TI - Ultrasound scan to detect acalculous cholecystopathy in immunocompromised hosts with unexplained fever. PMID- 10681741 TI - Fludarabine containing regimen followed by autologous peripheral blood stem cell transplantation in unselected patients with acute myeloid leukemia: a single center experience. PMID- 10681742 TI - [Modulated effects of prostaglandin E2 on endothelin production of alveolar macrophages in rats]. AB - Using radioimmunoassay, we studied the endothelin (ET) production of alveolar macrophages(AM) of the rats. The results showed that: (1) a basal amount of ET which was time-dependent (r = 0.7415, P < 0.01) was detected in supernatant of cultured unstimulated AM; (2) lipopolysaccharide (LPS), PMA, or A23187 could increase the ET production of AM (P < 0.01) (3) calmodulin antagonist W7 reduced the ET production of LPS or A23187-stimulated AM (P < 0.05, P < 0.01), but did not effect that of PMA-stimulated AM; protein kinase C inhibitor H7 attenuated the effect of PMA on ET production (P < 0.01), but did not effect LPS on ET production; (4) prostaglandin E2 (PGE2) inhibited the ET production of LPS stimulated (P < 0.05) and PMA-stimulated AM (P < 0.05); cyclooxygenase inhibitor indomethacin enhanced the effect of LPS on ET production (P < 0.01), but did not effect PMA on ET production. We conclude that AM is an important source of ET in the lungs both at physiologic and pathologic situation there are two pathways of signal transduction for factors stimulating ET production of AM, i.e., PKC dependent and Ca(2+)-calmodulin-dependent pathways; the autocrine PGEs from AM shows a negatively modulated effect on ET production of AM. PMID- 10681743 TI - [Identification of antigens shared between Schistosoma japonicum and Trichinella spiralis]. AB - Cross-reactivity was analysed between the antigen of T. spiralis muscular larvae and antigens of cercariae, liver stage schistosomula, 30 day male and female adult worms of S. japonicum by means of EITB technique. The result showed that the antigens of cercariae schistosomula, female worms except male worms, were recognized by sera from rabbits immunized with T. spiralis antigens or by sera from T. spiralis infected rabbits. The molecular weight of recognized antigens ranges from 15 to 100 kD, mainly from 50 to 70 kD. The immunized rabbit sera and the infected rabbit sera reacted with the same cross-antigens of S. japonicum. Among these schistosome stages, the cercariae showed the most cross-reactive bands, the second were the liver stage schistosomula, and 30 day female worms only showed a weak band which located at 97 kD position. Meanwhile, the anti-S. japonicum sera recognized several T. spiralis antigens which located at 34 kD and above, especially 35-38 kD. The results suggest that there are many antigens shared between T. spiralis and S. japonicum and the main cross-reactive antigens are found in the larva stages of S. japonicum and T. spiralis muscular larva. The cross-reactive antigens are different between various stages of S. japonicum and muscular larva of T. spiralis. PMID- 10681744 TI - [Detection of human cytomegalovirus by PCR in patients with gastrointestinal disease]. AB - Human cytomegalovirus (HCMV) genomes was detected by PCR in 51 patients' biopsy or operation specimens with gastrointestinal disease. The positive rate was 9.80%. At the same time, 41 patients' serum HCMV-IgM was detected by indirect immunofluorescence assay (IFA); the positive rate was 46.34%. Patients with cancer and colitis exhibited a higher HCMV infection rate than those without cancer and colitis. The results suggest that PCR is sensitive and specific in detection of gastrointestinal HCMV infection, and the examination of biopsy or surgical specimen is superior than that of IFA of serum HMCV-IgG. PMID- 10681745 TI - [Genotype distribution of hepatitis C virus in hepatocellular carcinoma tissue in Hunan province]. AB - Genotypes of hepatitis C virus(HCV) were detected by PCR using type-specific primer in 50 patients' hepatocellular carcinoma(HCC) tissue. The results showed that in the 50 HCC specimens, 30(60%) and 3(6%) were infected with the HCV type II and III, 5(10%) and 8(16%) with type II + III and type II + IV, 2(4%) with type II + I + III in combination, respectively. 2 cases were negative for HCV. These data suggest that HCV type II may be a predominant genotype related to hepatocarcinogenesis in Hunan Province, some cases of HCC may result from coinfection of HCV type II and other genotypes, and only few HCC be separately caused by infection of HCV type III. PMID- 10681746 TI - [The relationship between C-erbB-2 expression with cell proliferative activity and prognosis of nasopharyngeal carcinomas]. AB - C-erbB-2 and proliferating cell nuclear antigen (PCNA) were detected by immunohistochemical and in situ hybridization methods in nasopharyngeal carcinomas(NPC) and pericarcinomatous tissues(PCT). Some NPC cases were followed up for more than 5 years. RESULTS: The positive rates of C-erbB-2 protein and C erbB-2 mRNA expression were 87.8%, and 84.0%, respectively in NPC and 74.6% and 76.5%, respectively in PCT. There was a coexpression of C-erbB-2 protein and mRNA. The significant difference for PCNA staining intensity index(S II) existed in the vesico-nuclear and poorly differentiated types of NPC and in the C-erbB-2 staining cases of NPC. No correlation was found between the expression of C-erbB 2 protein and the clinical stage and metastasis as well as survival rate. CONCLUSION: The C-erbB-2 gene overexpression and cell abnormal proliferation are associated with the carcinogenesis and development of NPC. It is helpful to examine both C-erbB-2 gene expression and PCNA in NPC to evaluate the malignant degree and the effect of radiotherapy. PMID- 10681747 TI - [An ultrastructural observation of peripheral blood neutrophils in patients with hepatitis B]. AB - The abnormal changes of peripheral blood neutrophils in 13 patients with hepatitis B were observed under electron microscope. The morphological and structural changes of neutrophils included irregularity of shape, a large number of pseudopodia and phagocytic vacuoles, less electron dense granules, increased nuclear holes and vacuoles, etc. The present study provides the ultrastructural evidence of impaired neutrophil morphology and functions in such patients. PMID- 10681748 TI - [Significance of microvessel quantity in metastasis of invasive breast carcinoma]. AB - Microvessel quantity(MVQ), expression of CD44V6 and EGFR were studied in 48 patients with invasive breast carcinoma by CD31 immunohistochemistry. Significant differences of MVQ, CD44V6 and EGFR were observed (P < 0.01) between the groups with and without metastasis. The results suggest that angiogeneses and activation of tumor metastasis associated gene play an important role in tumor metastasis. PMID- 10681749 TI - [Influence of portal triad clamping on the intestine in pigs]. AB - A pig model of liver ischemia and intestinal congestion was made by portal triad clamping (PTC) for 45 minutes to investigate the influence of PTC on intestine. The results were as follows: 1. The level of cathepsin D was increased significantly after PTC as compared with the value before clamping (P < 0.05); 2. The endotoxin and lactic acid were significantly increased after PTC comparing with those of pre-clamping (P < 0.05, P < 0.01) and the control group (P < 0.05, P < 0.01); 3. The intestinal mucosa had a significant damage (P < 0.05) after PTC. The results indicate that PTC can result in intestinal mucosal lesion and enterotoxin absorption. Therefore, the protection of intestinal functions is important in hepatic surgery and preoperative bowel preparation should be done. PMID- 10681750 TI - [The effects of hypertonic saline/mannitol resuscitation on the myocardiac contractility and blood pressure of rabbits with hemorrhagic shock]. AB - Eighteen healthy adult male rabbits were hemorrhaged to shock and treated with hypertonic-hyperoncotic solutions in 3 groups (6 per group): hypertonic saline/dextran(HSD), hypertonic saline/mannitol(HSM) and hypertonic acetate/mannitol(HAM). The results showed that HSM. HAM and HSD could raise the left ventricular pressure(LVP), maximum change rate of LVP (+/- dp/dt max) and the arterial blood pressure of hemorrhagic shock rabbits quickly at small dose. The efficiency of HSM was better than that of HSD and HAM (P < 0.05), and the time of blood pressure rise was in accordance with that of LVP and +/- dp/dt max, which may partly explain the mechanism of shock resuscitation by hypertonic hyperoncotic solution. PMID- 10681751 TI - [Study on the immunoregulatory effect of Baoyuantang ([symbol: see text]) in patients with chronic hepatitis B]. AB - The T lymphocyte colong, formition (TL-CFU), the T lymphocyte subsets (CD3+, CD4+ and CD8+) and interieakin-2 membrane receptor (mIL-2R) secreted by peripheral mononuclear cells of 60 patients with chronic B hepatitis were studied. The results indicated that the activity of TL-CFU, mIL-2R and CD3+, CD4+, CD4+/CD8+ were decreased, and CD8+ was increased as compared with the normal controls. After treatment with Baoyuantang, the TL-CFU, mIL-2R, CD3+, CD4+ and CD4+/CD8+ increased and CD8+ decreased as well. It is suggested that the patients with the chronic B hepatitis with immune function defficiency, and the Baoyuartan treatment is presented immunoregulatory. PMID- 10681752 TI - [Infection of cryptosporidium in child patients with diarhea in Changsha]. AB - Fecal specimens were collected from 102 outpatients with diarrhea in Children's Hospital from March to October, 1997. The fecal smears were examined for cryptosporozoites with the rapid modified acid-fast two-step methods. The positive rate was 4.90%(5/102). There was no significant difference between female and male (P > 0.05). The positive rates of 1-month-, 5-year-, and 10-year old groups were 4.05% (3/74), 7.14%(1/14), and 7.14(1/14), respectively. We conclude that there is crytosporidium infection in child patients with diarrhea in different sex and age groups. PMID- 10681753 TI - [Dental fluorosis of brick tea type in rats]. AB - To observe the dental fluorosis of brick tea type, 48 white rats were fed by 245 mg.L-1 of brick tea, respectively. The rat model of brick tea type fluorosis were successfully established after 90 days. Naked eye examination of rats' incisors showed that brick tea and sodium fluoride (as positive control) groups all suffered from dental fluorosis. Using the dissecting microscope, the scanning electron microscope, and the energy spectrum apparatus, we also proved that the high concentration of brick tea could cause dental fluorosis. PMID- 10681754 TI - [Relationship between osteoporosis and metabolism of calcium and bone]. AB - Using metabolic balance test, bone mineral density measurement, animal models of osteoporosis, bone cell culture, we investigated the relationship between osteoporosis and the metabolism of calcium and bone. We found that the calcium intake was rather low in 20 healthy adults (480 mg per day); the average intestinal net absorption rate(%) of calcium in 124 healthy adult volunteers was 28.9(%), which increased gradually in recent 20 years (from 17.1% in 1978 to 40.6% in 1977), owing to improvement of dietetic structure. We conclude that the insufficiency of sex hormone and calcium are the main causes of osteoporosis. In bone cell culture, the whole process of bone remodelling was observed for 35 days continuously. Calcitonin damaging osteoclast was also investigated. Thus, we suggest new concept of bone remodelling coupling. PMID- 10681755 TI - [Mifepristone in the treatment of 47 ectopic pregnancy patients]. AB - OBJECTIVE: To investigate the effect and indication of mifepristone in treating ectopic pregnancy, 47 patients were divided into three groups. RESULTS: Twenty nine cases were effective; 18 cases were ineffective and operated on in which we observed the gestational termination, bleeding capacity, and pathological changes in fallopian tube. We found that the effect was related to the drug dosage, levels of beta-hCG in blood and the operating time. CONCLUSION: High dosage of mifepristone in treating ectopic pregnancy is convenient, safe, and has no side effects. It is suitable for patients with early ectopic pregnancy who have smooth life-sign, beta-hCG < 10 ng.ml-1, the mass in pelvic cavity < or = 5 cm, without acute belly-ache and pulsation of fetal heart in fallopian tube and who ask for the conservative treatment. PMID- 10681756 TI - [A study on HBV DNA pre C region A83 mutation in patients with hepatitis B]. AB - Mismatched polymerase chain reaction (MPCR) and restriction fragment length polymorphism assay(RFLP) were performed to detect the HBV DNA pre core A83 mutation in 97 patients with chronic HBV infection in Hunan. HBV DNA pre core A83 mutation were detected in 37 out of 97 patients (37/97,38.1%). It was found that the prevalence of HBV DNA pre core A83 mutation of the patients with severe hepatitis and with CAH(chronic active hepatitis) were significantly higher than those with CPH(chronic persistent hepatitis) and carriers of HBV, and the patients with elevated transaminase (ALT > 70U) or positive HBeAb were significantly higher than those with normal transaminase (ALT < 70U) and with positive HBeAg, respectively. The results suggest that the HBV DNA pre core A83 mutation is associated with active and severe liver tissue in patients with hepatitis B. PMID- 10681757 TI - [Changes of tissue oxygenation and relationship between oxygen delivery and consumption in patients with poor cardiac function undergoing valve replacement]. AB - To study the relationship between oxygen delivery(DO2) and consumption(VO2) and tissue oxygenation in patients during cardiac valve replacement, DO2, VO2, arterial blood lactate (ABL) SvO2 were measured. The results showed a poor correlation between DO2 and VO2 and a normal tissue oxygenation before CPB. There was a good positive correlation between DO2 and VO2 within 20 h after the end of CPB(P < 0.01), and changes of ABL and SvO2 showed tissue hypoxia. The level of ABL in patients showing O2 supply-dependency was lower, and SvO2 was higher than those in 15 patients showing O2 supply-independency, respectively (P < 0.05). These data suggest that O2 supply-dependency was existed after the end of CPB, and there was a good correlation between O2 supply-dependency and ABL. It might not exclude tissue hypoxia in O2 supply-independency and normal level of SvO2. PMID- 10681758 TI - [Dynamic enhanced MR imaging of intraspinal tumors]. AB - The dynamic enhanced MR imaging of 37 patients with intraspinal tumors were performed. We found that the enhancement peak within 90s was shown only in hemangioblastoma, within 180s was shown in most astrocytomas, meningiomas, and metastases. The maximum contrast enhancement ratio (MCER) of neurilemmoma is higher than that of meningioma(P > 0.05). We conclude that the contrast enhancement ratio-time(CER-T) curve, the enhanced degree, and pattern of intraspinal tumors have the significant value for the histologic diagnosis and the calculation of vascularity. PMID- 10681759 TI - [Serum HBV DNA detected by polymerase chain reaction with dUTP/uracil-DNA glycosylase]. AB - The ability of PCR reagent containing dUTP/uracil-DNA glycosylase for controlling carry-over contamination of PCR products was explored. All of 204 sera taken from hepatitis patients were used for HBV DNA detection by PCR with PCR-dUTP/UDG reagent in comparison with that without dUTP/UDG. The results showed that its efficiency of controlling contamination was excellent. At least, contamination of 100 ng PCR products was got rid of. The corresponding rate of HBV DNA detection by PCR-dUTP/UDG in combination with dot hybridization using digoxin-labeled HBV probe was as high as 89.32%, higher than that (81.45%) of PCR without dUTP/UDG plus dot hybridization(P < 0.05). It suggests that PCR-dUTP/UDG method could prevent PCR products from contaminating and increase accuracy and specificity of PCR amplification. PMID- 10681760 TI - [Parkinson's disease and dementia]. AB - Neuropsychological tests were carried out in 30 patients with Parkinson's disease and 26 healthy controls. The results showed that the average scores of HRB(A)-RC test were above the limited scores, 45.45%-86.36% cases being over the limited scores (P < 0.01), except speech sound perception test. Damage degrees (DQ) from the least to the most serious were 63.64%. WAIS-RC test and WMS test showed that the average scores of IQ and MQ in patients were lower than those in the normal. The difference of IQ, MQ scores between patients and controls was very significant (P < 0.01). The damaged cases of IQ, MQ from the least to the most serious were 20% and 60%, respectively. This indicated that the patients with Parkinson's disease were obviously damaged in intelligence and memory. In this article, there were 6 cases (20%) fitted the diagnostic standard of dementia, among them, 4 were subcortical dementia and 2 cases were complex dementia. We found that the differences between the course and IQ, MQ were very significant (P < 0.05), but not between the course and DQ. It is implied that in the very early stage, the patients could appear abnormal in neuropsychology, and the intelligence and memory may be damaged more seriously in longer course. The result suggests that the early diagnosis and prompt intervention is important to prevent the occurrence and advance of dementia. PMID- 10681761 TI - [Antitumorigenic immunocompetence of alveolar macrophages in patients with lung cancer]. AB - OBJECTIVE: The antitumorigenic immunocompetence of alveolar macrophages(AMs) in patients with lung cancer was investigated. METHODS: Twenty-two patients with lung cancer and 18 control subjects underwent bronchoalveolar lavage (BAL) by fiberoptic bronchoscopy to collect BAL fluid (BALF). AMs in the BALF isolated by adherence on plate surface were incubated for 24 hours at 5% CO2, 37 degrees C in medium with or without granunocyte-macrophage colony stimulating factor (GM-CSF). The cell free supernatants (SP) and AMs were harvested, respectively, the activities of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), superoxide dismutase (SOD), and soluble interleukin 2 receptor in BALF and SP were examined by copper plating cadmium reduction, radioimmunoassay, xanthine oxidase assay, and ELISA, respectively. mRNA expression of inducible nitric oxide synthase in AMs was detected by reverse transcription-polymerase chain reaction(RT-PCR). RESULTS: (1) The activity of NO was lower in BALF of the tumor bearing lungs than that of the nontumor-bearing lungs and control groups; the activities of TNF alpha, SOD, and sIL-2R were not statistically significant in BALF between patients with lung cancer and control groups. (2) In supernatants, the activities of NO and TNF alpha were lower in the tumor-bearing lungs than in nontumor-bearing lungs and control groups; sIL-2R concentration of the lung cancers was higher than that of control groups. (3) After AMs were stimulated by GM-CSF, the activities of NO, TNF alpha, and SOD were increased and that of sIL 2R was decreased in SP. (4) mRNA expression of iNOS was demonstrated in AMs of both groups. However, in comparison with beta-actin, the intensity of mRNA expression of iNOS in AMs of lung cancer patients was lower than that of control groups. CONCLUSION: There are some deficiencies in the function of AM at the region of tumors and antitumorigenic immunocompetence in the patients with lung cancer. PMID- 10681762 TI - [Expression of telomerase in acute leukemia]. AB - To understand the expression patterns of telomerase activity in different types of acute leukemia(AL) and during remission state, the mono-nuclear cells of different bone marrow samples were isolated by desity gradient centrifugation. Telomerase activity of the samples was assayed by telomeric repeat amplification protocol(TRAP) with the cell extracts and the TRAP products were resolved in PAGE. The results showed that the telomerase activity was at higher levels in the acute leukemia compared with the normal controls. Of the acute nonlymphocytic leukemia(ANLL), the telomerase activity presented as M2a > M5b > M1 > M3b. Of the acute lymphocytic leukemia(ALL), the activity presented as L1 > L2. The activity assayed in M2a and L2 remission was as high as that in the normal group. It indicated that the expression of telomerase was enhanced differently among the subtypes of ANLL and ALL, and was down-regulated during the AL remission. It implicates that the expression patterns of telomerase activity in AL is associated with the different proliferation states and differentiation properties of the AL cells. PMID- 10681763 TI - [A preliminary report of antiscar formation effect by subconjunctival injection of mitomycin C before trabeculectomy in glaucoma]. AB - To improve the function of filtering blebs and increase the successful rate of the trabeculectomy, mitomycin C injection 0.02 mg.ml-1 was given subconjunctivally at the operative site before trabeculectomy in 32 patients (38 eyes) with moderate or late glaucoma. All patients were followed up for 6 to 12 months. The results showed that the rate of functional blebs were 93.7%. The successful rate of the operation was 94.7%. The common postoperative complication was shallow anterior chamber (7.9%). These findings indicate that the subconjunctival injection of mitomycin C before operation is a safe, effective method for antiscar formation. PMID- 10681764 TI - [Changes of blood supply from portal system after transcatheter arterial chemoembolization in huge hepatocellular carcinoma]. AB - OBJECTIVE: Blood supply changes of the portal venous system to the tumor mass before and after transcatheterarterial chemoembolization(TACE) in a series of 22 patients with huge hepatocellular carcinoma were monitored to study the necessity of intervention via portal vein. METHODS: Twenty-two selected patients with huge hepatocellular carcinoma were enrolled in the study. Elscint double helical CT was used to scan liver during the maximum concentration of contrast medium in portal venous system. CT Angiography and MIP(Maximum Intensity Projection) were used to image formation of the portal venous system. RESULTS: The technique of scan and image formation were excellent to evaluate the attribution of portal venous blood supply to HCC. The portal venous blood supply was mainly the tumor periphery distribution (P < 0.01). After treatment of TACE, the portal venous blood supply to tumor periphery increased significantly (P < 0.05), and some did appear signs of enriched blood supply around the tumor. Negative correlation between the degree of reduction in tumor and portal venous blood supply to the tumor periphery before TACE was evident. CONCLUSION: The attribution of portal venous blood supply to HCC was in the tumor periphery, and regression of tumor after TACE was affected by portal venous blood supply to the tumor, thus it is reasonable to suggest interventional therapy via portal vein. PMID- 10681765 TI - [Plasma levels of lipids, lipoproteins and apolipoproteins affected by endogenous testosterone]. AB - Plasma testosterone(T), estrodiol(E), total cholesterol(TC), triglyceride(TG), high density lipoprotein-cholesterol(HDL-C), HDL2-C, HDL3-C, lipoprotein(a)[Lp(a)], apolipoprotein(Apo) AI and B100 were determined in 201 subjects, among them 102 patients with coronary heart disease(CHD) and 99 healthy subjects. It was observed that plasma T level was correlated negatively with TG(r = -0.47, P < 0.01) and Lp(a) (r = -0.163, P < 0.05), and positively with HDL3-C(r = 0.328, P < 0.01) and Lp(a)(r = -0.163, P < 0.05), and positively with HDL-3(r = 0.328, P < 0.01) and HDL3-C(r = 0.328, P < 0.01). Meanwhile, plasma levels of TG (1.47 +/- 1.16 mmol.L-1) and Lp(a)(25 +/- 17.8 g.L-1) were significantly higher in the subjects with low plasma T level (< 270 ng.ml-1) than those in the subjects with normal plasma T level (1.10 +/- 0.74 mmol.L-1 and 17 +/- 15.9 g.L 1, respectively), while plasma levels of HDL-C (1.31 +/- 0.30 mmol.L-1) and HDL3 C(0.88 +/- 0.22 mmol.L-1) were significantly lower in the former than in the latter (1.44 +/- 0.33 mmol.L-1 and 1.02 +/- 0.26 mmol.L-1, respectively). The results support that low plasma T level may be a risk factor for coronary heart disease, which may relate to the changes of plasma lipoproteins. PMID- 10681766 TI - [Clinical analysis of PIH with abnormal liver function in 66 patients]. AB - Sixty-six patients of severe pregnancy induced-hypertension(PIH) with abnormal liver function were studied. The results showed that the incidence of abnormal liver function in severe PIH was 20.62%, which was significantly higher than that in moderate PIH and closely related to the course of the disease. The incidence of abnormal liver function in patients who were sick longer than 1 month was much higher than those with short course (P < 0.05). The liver function abnormality in PIH mainly showed the elevation of SGPT activity, increase of serum bilirubin was rarely seen. Besides these, the incidences of the fetal distress and postpartum hemorrhage in patients with abnormal liver function were significantly higher as compared with those having normal liver function (P < 0.05). PMID- 10681767 TI - [Protective effects of shenmai cardioplegia on intracardiac operative patients with cardiopulmonary bypass]. AB - Forty intracardiac-operative patients were divided into two groups randomly. Twenty patients were received by the improved St. Thomas Crystal Blood (1:4) Cardioplegia(Group A); 20 by the Shenmai liquid and improved St. Thomas Crystal Blood(1:4) Cardioplegia(Group B). During perioperation, the blood samples and myocardial tissue were collected, and clinical observations were recorded. The results showed that in Group B, contents of myocardial specific isoenzyme of creatine kinase(CK-MB), lactate dehydrogenase(LDH), malonyldialdehyde(MDA), and myocardial Ca2+ were significantly lower, the contents of superoxide dismutase(SOD) were, significantly better than that in Group A; and the histological findings showed less damage in Group B than in Group A. We conclude that Shenmai Cardioplegia can increase the myocardial protection of the intracardiac-operative patients. The possible mechanisms are that Shenmai liquids inhibit the production of oxygen free radicals and calcium paradox. PMID- 10681769 TI - [The diagnosis and treatment of nipple discharge in 253 cases]. AB - Among 253 cases of nipple discharge, 96 cases of them were diagnosed as intracanalicular papilloma(37.9%), 73 cases as cystic disease of breast(28.9%), 41 cases as mammary duct ectasia (16.2%), 35 cases as breast cancer(13.8%) and 8 cases as acute suppurative mastitis(3.2%). The masses beneath areolar region were smaller those outside areola. Finding of cancer cells or suspicious cancer cells through cytologic examination of nipple discharge smear and breast mass puncture usually had important significance. In addition, near infrared ray scanning shows high rate of correct diagnosis; estimation of carcinoembryonic autigen(CEA) in nipple discharge is beneficial to the early diagnosis of malignant diseases. PMID- 10681768 TI - [The second phase clinical observation of anti-radiation effect by superoxide dismutase]. AB - Multiple center randomized controlled double blind clinical trait was conducted to evaluate the anti-radiotherapy effect by SOD (produced by Hunan Biochemical Work) in 159 patients. Injection of 4000U SOD immediately after receiving radiotherapy significantly reduced the occurrence rate of skin, oral mucosal, pelvic visceral and systematic adverse reaction, only the reduction of leukopenia did not reach the statistical significant level. No adverse effect of SOD injection was observed. The results suggest that SOD is a safe and effective agent to attenuate the radiotherapy reactions. PMID- 10681770 TI - [Detection of 17-ketosteroid and 17-hydroxycorticosteroid in urine after separation and collection by foam rubber]. AB - Using foam rubber 17-ketosteroid (17-KS) and 17-hydroxycorticosteroid (17-OHCS) were separated and collected in urine. 17-KS and 17-OHCS were eluted down from the foam rubber with alcohol, colour reaction can be respectively produced by m dinitrobenzene and phenyhydrazine hydrocholoride, then both of them were detected by spectro-photometry. The standard deviation of this method was +/- 0.032, and its minimum detectable concentration was 0.01 microgram.ml-1. The recovery rate of 17-KS and 17-OHCS estimation was from 99.0% to 101.0%, with co-efficients of variation less than 6.23%. PMID- 10681771 TI - [Determination of glycosylated hemoglobin in human erythrocytes by fast protein liquid chromatography system]. AB - The chromatography method for analysis of glycosylated hemoglobin(HbA1c) in human erythocytes was established by using a fast protein liquid chromatography system(FPLC) and a Mono S column. The average intrabatch and interbatch coefficients of variations (n = 3) were 2.5% and 4.8%, respectively. There was a positive correlation between the results of FPLC and routine microcolumn method (r = 0.413, P < 0.02). Using FPLC method, the HbA1c levels in festing blood of 54 normal donors and 26 diabetics on admission were 4.6 +/- 1.19(%) and 10.2 +/- 3.45(%), respectively. A significant difference was found statistically between the results of two groups (t = 10.8, P < 0.01). PMID- 10681772 TI - [Effect of ultraviolet irradiation on immune function in rabbits]. PMID- 10681773 TI - [Analysis of risk factors, prevention and therapy of fungal infection on brain injuries]. PMID- 10681774 TI - [Effects of sevoflurane on hemodynamics of open heart valve replacement surgery]. PMID- 10681775 TI - [Relation between spirochetes and retrograde pulpitis]. PMID- 10681776 TI - [Relation between ABO blood-group system and Helicobacter pylori infection]. PMID- 10681777 TI - [Clinical uses of autologous blood transfusion]. PMID- 10681778 TI - [Mitral valve prosthesis implantation complication: thrombosis]. PMID- 10681779 TI - [Ultrasonic diagnosis for orthopedic (corrective) transposition of the great artery with complex cardiac deformity. A case report]. PMID- 10681780 TI - [Expression of connexin genes in the liver of different stages of embryo development]. AB - To explore the relationship between the connexin (Cx) gene expression and cellular differentiation in the liver of different embryonic stage, we studied the expression of Cx genes in the liver of different embryonic ages by Northern blot hybridization technique, using the series of Cx gene as the molecular probes. The expression of Cx genes exhibited organ specificity, Cx26, Cx32, and Cx43 were expressed in the liver while others had no signal. Cx26, Cx32, and Cx43 genes exhibited different expression models in different developmental stages of the fetus. Cx43 did not express in human liver after birth, but it expressed in the fetal stage. The results suggest that the expression of Cx genes plays an important role in the organ development and may be a key gene to regulate some differentiational events associated with cell growth and organ development. PMID- 10681781 TI - [Expression of CD44V6 and nm23-H1 in thyroid papillary adenocarcinoma and lymph node metastasis]. AB - Expression of CD44V6 and nm23-H1 in thyroid papillary adenocarcinoma (TPC) was studied in 84 patients by immunohistochemistry. The positive rates of CD44V6 and nm23-H1 were 63.1% and 48.8% respectively. An increased CD44V6 and a decreased nm23-H1 positive rate in patients with lymph node metastasis (LNM) were observed (P < 0.01). Negative correlation was existed between overexpression of CD44V6 and low-expression of nm23-H1 (P < 0.01). The results suggested that CD44V6 and nm23 H1 play important role in LNM of TPC and disordered expression of CD44V6 and nm23 H1 are involved in the process of LNM. PMID- 10681782 TI - [Effects of mild hypothermia on Na(+)-K+ ATPase and lipid peroxidation in canine brain tissue following cardiac arrest and resuscitation]. AB - The effect of mild hypothermia on Na(+)-K+ ATPase and lipid peroxidation in canine brain tissue following a 18-minute cardiac arrest and resuscitation for 8 hours were studied. Mild hypothermia improved the restoration of the activity of Na(+)-K+ ATPase, LDH, protect the activity of SOD, decrease the loss of GSH, but not completely blocked the ischemia reperfusion induced lipid peroxidation. PMID- 10681783 TI - [Microtubule-associated protein30 (MAP30) and ubiquitin detected in neurofibrillary tangles of Alzheimer's disease brains]. AB - Using immunohistochemical technique, the distribution of immunoreactive products of antiubiquitin and MAP30 in the neurons of hippocampus of Alzheimer's disease (AD) brain were studied. Nurofibrillary tangles (NFT), dystrophic neurites in plaques and neuropil threads were labeled by the antibody of ubiquitin. NFT and pretangle neurons (PTN) were stained by the antibody of MAP30, but not detectable signal in axons or distal dendrites. The neurons immunostained with antibodies to ubiquitin and MAP30 were found in all the areas of CA1, CA2, CA3 and CA4 of hippocampus of AD. These neurons labeled by 2 antibodies used in this study were distributed more in the CA1 and CA2 than those in CA3 and CA4. The results indicate that ubiquitin is seen associated when NFT is already formed, and MAP30 may be a novel antigenic component of NFT in Alzheimer's disease. PMID- 10681784 TI - [Protection of hypertonic saline/mannitol to thymic cell apoptosis induced by endotoxin (LPS) in mice]. AB - To observe the effect of HSM (hypertonic saline/mannitol) on thymic cell apoptosis induced by endotoxin (LPS) in mice, twenty-four mice were divided into three groups (8 mice/each group): 1. Control group, each mouse was intraperitonealy injected with normal saline; 2. LPS group, each mouse was intraperitonealy injected with LPS; 3. LPS + HSM group, each mouse was intraperitonealy injected with HSM before and after injection of LPS. The results showed that the percentage of apoptosis of HSM + LPS group was lower than that of LPS group (P < 0.01); the ladder of nucleosomal DNA fragments by agarose gel eletrophoresis of HSM group was not typical as compared with that of LPS group. It is suggested that HSM might protect the apoptosis of mouse thymic cell induced by LPS and may improve the immune function. PMID- 10681785 TI - [The expressions of p16, CDK4 and PCNA proteins in trophoblastic tumors]. AB - OBJECTIVE: To evaluate the relationship between the regulatory factors in G1 phase of cell cycle and the cacrinogenesis of the trophoblastic cells. METHODS: The expressions of p16, cyclin-dependent kinase 4 (CDK4), proliferation cell nuclear antigen (PCNA) proteins in 18 cases of trophoblastic tumors, 30 cases of hydatidiform mole and 30 cases of normal villi were studied by immunohistochemical methods. RESULTS: The expressions of p16 protein between malignant trophoblastic tumors and normal villi were significantly different (P < 0.05). PCNA positive rate in p16 positive samples were markedly lower than that in negative samples (P < 0.05). The expressions of CDK4 among normal villi, hydatiform mole and trophoblastic tumors were not significantly different. Patients with positive p16 expression had a higher 3 year survival rate. CONCLUSION: p16 protein may inhibit trophoblastic cells proliferation, and p16 gene mutation may be an important factor in carcinogenesis of trophoblastic cells and proliferation out of control. P16 protein expression examination is helpful in prediction of patients prognosis. PMID- 10681786 TI - [Determination of cyanide in whole blood by completely differential spectrophotometry]. AB - This paper introduced a method for the determination of cyanide in whole blood by completely differential spectrophotometry with pyridine-pyrazolone method after stabilization by addition of silver sulfate and separation by distillation. The analytical results showed a precision 5.4% (n = 6) for the determination of 0.068 microgram.ml-1 CN- in whole blood. The average recovery was 97.4%. This method was successfully used for the determination of cyanide in whole blood of non smokers and smokers. PMID- 10681787 TI - [Four-year follow-up and molecular epidemiologic investigation of intrafamilial transmission of HCV infection]. AB - OBJECTIVE: To study the intrafamilial transmission of HCV infection. METHODS: Four-year follow-up and molecular epidemiologic investigation of HCV infection in a village was carried out. The diversity of HCV core region was analysed by reverse transcription nested polymerase chain reaction, Okamoto genotyping method, and single strand conformation polymorphism analysis (SSCP). RESULTS: The incidence of positivity of anti-HCV and -HCVRNA raised from 21.30% (75/352) to 43.18% (152/352). There were 42 families with two or more members serological positive for anti-HCV and/or -HCVRNA. There were a 3.46-fold increase of intrafamilial propagation of HCV infection in the past 4 years. HCV genotyping by Okamoto method showed that 90.67% (136/150) belonged to Type 1b/II, two cases (1.33%) belong to type 2a/III and three cases (2.00%) were co-infection of both types. Comparison of SSCP patterns revealed that the SSCP of HCV cDNA was identical among members of the same family, but the single strand conformation of HCV cDNA exhibited polymorphism patterns in different families. CONCLUSION: The results indicate that the cause of marked increase of HCV infection rate in this village in a 4-year period is mainly because of intrafamilial propagation. SSCP is more feasible for molecular epidemiologic investigation of HCV infection than genotyping. PMID- 10681788 TI - [Thiamine status of university teacher families in Changsha]. AB - To evaluate the thiamine status of the population consuming the refined cereals and its products, we studied thiamine concentration in 23 food samples and human urine, and three-day dietary survey was conducted. The results showed that the thiamine content in the staple food was much lower than that in the raw material. The thiamine intake from diet was below to 80% RDA in 42.8% investigated families and the thiamine energy ratio was lower than 0.5 mg/4186.8 kJ in 71.5% of them. For the ratio of urinary tiamine (mg)/urinary inosine (g), 51.3% subjects exhibited decreased excretion, especially in the school-age children. We conclude that those people who take the refined cereals and its products as a staple food, is in the risk of thiamine deficiency. PMID- 10681789 TI - [Study on the characteristics of personality, emotion and the plasma arginine vasopressin levels in patients with Gan Yang Shang Kang Zheng]. AB - Eighty patients with Gan Yang Shang Kang Zheng (GYSKZ) and the related Zheng were questioned with three psychological scales: Type A Behavior Inventory, STAI and SDS and plasma AVP (arginine ressopressin) levels were determined by RIA. The TAB, S-AI, T-AI scores and plasma AVP levels of the patients with GYSKZ increased consistently and the changes of all parameters in different diseases in the same Zheng were similar. The results suggest that in patients with GYSKZ, type A behavior is the dominant personality and anxiety is the chief emotional disorder. Patients with GYSKZ are in the state of high level of psychological stress. Patients with GYSKZ and GHSYZ have some common pathophysiological basis. PMID- 10681790 TI - [Measurement of the strength of pronation and supination forces and its significance in radius fracture displacement]. AB - The strength of pronation and supination was measured by "Forearm-Rotate Dynamometer" on 524 male and female students ranging in age from 16 to 32. The study showed that the strength of supination was 13% greater than that of pronation for 91.3% of all the tested subjects (1048 sides). Based on this finding, the pattern of displacement fracture of the middle lower segment of the radius was discussed. It was concluded that the proximal fragment displacement will show supination, adduction and flexion when a proximal fragment displacement occurs. PMID- 10681791 TI - [Experimental study on hypoglycemic effect of YTY granules]. AB - OBJECT: To perform experimental pharmacodynamic evaluation of a new TCM antidiabetic drug Yitangyin (YTY). METHODS: Experimkental streptozotocin induced rat diabetes models were subjected YTY treatment and the hypoglycemic effect was evaluated. The proved TCM Yusanxiao (YSX) was used as contrast. RESULTS: The low, middle and high dose YTY groups exhibited significant hypoglycemic effects on the model rats. The efficiency was comparable with YSX. CONCLUSION: The TCM YTY granule is an effective hypoglycmeic agent. PMID- 10681792 TI - [The protective effects of cardiac ischemic preconditioning on lung in cardiac operation with cardiopulmonary bypass]. AB - Twenty direct vision intracardial operation patients were divided into two groups randomly. After cardiopulmonary bypass, ten patients were treated with myocardial ischemic preconditioning. The aorta were clamped for 3 minutes and released for 3 minutes (Group IP). Another ten patients were not treated with ischemic preconditioning (Group C), only underwent 6 minutes cardiopulmonary bypass. Then the aorta were clamped and intracardial operation were done. The left atrium blood and lung tissue were collected just after thoractomy and half an hour after cardiac reperfusion in both groups. RESULTS: (1) The numbers of polymorphonuclear (PMN) of the two groups were increased significantly after cardiopulmonary bypass (P < 0.01). (2) The number of PMN and SOD, PaO2 contents were significantly higher in Group IP than in Group C (P < 0.05). (3) The numbers of PMN in lung interstitum under microscopy were less in Group IP than in Group C. (4) MDA contents were less in Group IP than in Group C (P < 0.05). (5) Histological finding showed less damage in Group IP than in Group C. It is evident that cardiac ischemic preconditioning could protect lung against ischemia reperfusion injury. The possible mechanisms are that ischemic preconditioning inhibites the accumulation and activation of PMN in lung tissue and reduces the production of oxygen free radicals. PMID- 10681793 TI - [Changes of blood lipids and apolipoproteins in patients with silent myocardial ischemia]. AB - Serum lipid profiles and apolipoproteins were measured in 148 patients with silent myocardial ischemia (SMI) and 30 healthy control subjects comparable in age. The serum lipid profiles and apolipoproteins were abnormal in all types of SMI which were more significant in type II and III. The extent of ST segment depression in ECG were positively correlated with serum TG, TC, LDL, B100, B100/A1, Lp(a); and negatively correlated with A1, HDL1, HDL2, HDL-C/TC. Multiple factor stepwise regression analysis revealed that the increased concentration of serum TG, LDL, and B100/A1 ratio and decrease of HDL2-C are independent risk factors in SMI. PMID- 10681794 TI - [Comparative study of left ventricular morphology and function of patients with left ventricular systolic and diastolic heart failure on echocardiography]. AB - Left ventricular pathology and pathophysiological characteristics of fifty patients with left ventricular diastolic heart failure (LVDHF), and 35 patients with left ventricular systolic heart failure (LVSHF) diagnosed by clinical manifestations and radionuclide ventriculography, were examined by echocardiography and 20 healthy persons were served as control group. Left atrial diameter (LAD), interventricular septum thickness (IVST) and posterior wall thickness (PWT) and left ventricular mass index (LVMI) were significantly increased in the patients with LVDHF, while left ventricular diameter (LVD) was significantly increased in the patients with LVSHF. Left ventricular ejection fraction (LVEF), cardiac index (CI) were significantly decreased in the patients with LVSHF. Early peak filling velocity (EPFV), atiral peak filling velocity (APFV), deceleration of early peak filling velocity (DC) and isovolumic relaxation time (IRT) in the patients with LVDHF and LVSHF observed no significant difference. PMID- 10681795 TI - [Clinical study on left ventricular diastolic heart failure in the aged patients]. AB - Forty aged patients with LVDHF and 30 with left ventricular systolic heart failure (LVSHF), confirmed by radionuclide ventriculography were studied by clinical, echocardiography and treadmill test. Twenty healthy aged persons were served as control group. Compared with LVSHF, short duration and less severe manifestations of heart failure in LVDHF were observed. Compared with the control, left atrium was enlarged but not the left ventricular chamber, and LV wall was thickened in LVDHF, while enlargement of LV chamber was only found in LVSHF. However, LV systolic function was remained normal in LVDHF. The exercise tolerance was decreased in both LVDHF and LVSHF groups, but more marked in LVSHF. PMID- 10681796 TI - [Changes of plasma testosterone level in male patients with coronary heart disease]. AB - The plasma testosterone (T), estrodiol (E), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), HDL2-C, HDL3-C, apolipoprotein (apo) A1 and B100, and lipoprotein (a) [Lp(a)] were measured in 118 male patients with coronary heart disease (CHD) and 53 sex and age-matched healthy controls. The results showed that plasma T, HDL-C, HDL3-C levels and HDL C/TC ratio were significantly lower and Lp(a) level was significantly higher in patients than in the controls. After adjusting body mass index, blood pressure and fasting blood glucose, the difference of plasma level between patients and controls persisted. Plasma T level correlated positively with the HDL-C, HDL3-C level and HDL-C/TC ratio and negatively with systolic blood pressure, apoB100 and Lp(a) level. Logistic regression was used to analyze the data, and this indicated that decreased T and HDL3-C levels, and increased Lp(a) level were the independent risk factors of CHD. PMID- 10681797 TI - [The application of low tidal volume pressure-controlled ventilation in patients with acute respiratory distress syndrome]. AB - Low tidal volume pressure-controlled ventilation therapy was used in 32 patients with ARDS. There were resulted a lower peak inspiratory pressure (PIP) level than that of control group, thus a decline in incidence of overventilation and the overventilation induced mortality. The results suggest that low tidal volume pressure-controlled ventilation may consider as preferred ventilation mortality in management of ARDS. PMID- 10681798 TI - [Prospective study on the correlation factors of fetal macrosomia]. AB - The increase of body weight in pregnancy women, 50 g oral glucose challenge test (50 g GCT), pregnant women and umbilicus vein blood insulin (INS), growth hormone (GH), glucose and neonate weight were measured in 110 pregnant women and their neonates were measured to explore the correlation factors of fetal macrosomia. The incidence of macrosomia was higher in pregnant women whose increased weight > or = 15 kg than those < 15 kg. 50 g GCT plasma glucose showed positive correlation with neonatal weight. Which was higher in the group of the fetal macrosomia than of the normal neonates. The results suggest that both the increasing weight of the pregnant women and the 50 g GCT are related parameters to predict fetal macrosomia. To prevent fetal macrosomia the diet should be limited in those who have had a high increasing body weight in pregnancy and appropriate intervention is required to those whose 50 g GCT are positive. PMID- 10681799 TI - [Detection of Toxoplasma gondii DNA in the autopsy cases of abnormal birth and teratosis by polymerase chain reaction]. AB - Toxoplasma gondii DNA in the autopsy tissues of 72 cases of the premature, stillbirth and congenital malformation was detected using polymerase chain reaction (PCR). The specific amplified products were found in 9 cases, i.e. one case of premature, 3 cases of stillbirth, one case of hydrocephalus and 4 cases of other teratosis. The positive rate was 12.5%. Ten control samples were negative. The results showed that Toxoplasma gondii infection was present in the disease cases. The finding indicates that the infection of Toxoplasma gondii may be one of the main causes of the abnormal birth and the congenital teratosis. PMID- 10681800 TI - [Internal fixation with mini-titanium plate for midface fractures]. AB - Thirty four patients with midface bone fractures were fixed by rigid internal fixation (RIF) with min-titanium plate. An optimum exposure is provided for the fracture sites of zygomatic, zygomatic arch, orbital floor and maxilla with the coronal incision and adjuvant incisions. The approach was of benefit to accurate anatomic reduction and fixation of the fracture segments. RIF with min-titanium plate provided three-dimensional stability. The fixation of zygomatic-frontal crevice and zygomatic maxillary crevice were the key for obtaining sufficient stability, and at least two fixation screws were required above zygomatic-frontal crevice. PMID- 10681801 TI - [Research in the relationship between Chlamydia and ureaplasma and infertility]. AB - Samples taken from cervix and male urethra of 768 infertilitas patients were studied, and samples taken from 226 normal fertile males and females served as control. The fluorescent monoclonal antibody technique and cell culture method were used for Chlamydia trachomatis (Ct) and microbiological culture method was used for Ureaplasma urealyticum (Uu) detection. The positive rates of Ct and Uu infections were 28.64% (220/768) and 36.59% (281/768) in infertilitas group, 5.2% (14/266) and 14.28% (38/266) in control group respectively. There was significant difference between the 2 groups (P < 0.01). The results suggest that infection with Ct or Uu is one of the factors causing infertility. PMID- 10681802 TI - [Etiology and clinical analysis of epididymal mass]. AB - The etiology and clinical pathological analysis of 147 cases of epididymal mass were presented. The result showed that the most common mass in inflammatory (66%), and the majority was tuberculosis, the next was cystic diseases (30%). However, neoplasm was relatively less and malignant mass was rare. The diagnosis can be established by careful history, physical examination, and BUS. The selective surgery depends on the nature of diseases, the demand of the patients and the effect of conservative therapy. PMID- 10681803 TI - [Relationship between clinical manifestation and pathology in pediatric renal diseases]. AB - To elevate the clinical diagnosis and pathological compatibility of pediatric kidney diseases. Pathological changes from 101 cases of pediatric renal diseases were studied, covering 13 diseases entities, including glomerulo-nephritis, simple nephrotic syndrome, nephritic nephrotic syndrome etc. The technique of tru cut biopsy under the guidance of B ultrasonogram was used in renal biopsy. The renal puncture biopsy was 100% satisfactory. Pathological changes involved 12 pathological types. MsPGN were the most commonly seen (39.6%), followed by MPGN and ICPGN (12.87% and 10.89%) respectively. It was found that children with similar clinical manifestation may possess different pathological changes, while the same pathological entity may present multiple clinical manifestation. The results suggest that renal biopsy pathological diagnosis is important in the diagnosis, treatment and prediction of prognosis in pediatric renal disease. PMID- 10681804 TI - [Preliminary study on Chlamydia pneumoniae pneumonia]. AB - In order to know the incidence of Chlamydia pneumoniae (strain TWAR) pneumonia and its clinical features, 93 patients with pneumonia and 93 matched patients with non-respiratory diseases were studied. TWAR antibodies (IgG and IgM) were detected by microimmunofluorescence (MIF) test. The results showed that 19.4% (18 cases) patients with pneumonia were TWAR pneumonia, in which 10 cases accompanied by bacteria infection and 7 cases being simple TWAR pneumonia. There were no significant differences in clinical features between TWAR pneumonia and non-TWAR pneumonia, except dry and moist rales. These data showed that the occurrence percentage of TWAR pneumonia in patients with lung cancer was higher than that in patients with the other respiratory diseases. This study suggests that there are TWAR pneumonia in China. PMID- 10681805 TI - [Ultrasonographic rating score in evaluation of pathological severity and timing of operation for acute cholecystitis]. AB - A rating score of ultrasonography to evaluate the pathological severity and timing of operative intervention of acute cholecystitis was designed. The ultrasonic rating score consisted of 7 items with were sensitive and specific in diagnosis of acute cholecystitis, accorded with the findings during operation. Emergency cholecystomies performed on patients with score < or = 10, 11-18 and > 18 points were 1/30, 4/42 and 7/13 respectively, followed by definite procedures. The results suggest that the ultrasonographic rating score is an useful approach in ultrasound diagnoses and timing of operation for acute cholecystitis. PMID- 10681806 TI - [Distribution of Mg2+ and Ca2+ in serum and lymphocyte of the patients with arrhythmia]. AB - The distributions of Mg2+ and Ca2+ in serum and lymphocyte from 51 arrhythmic patients (15 cases of atrial premature beat, 12 cases of atrial fibrillation, 24 cases of ventricular premature beat) and 30 healthy subjects were detected by flame atomic absorption spectrophotometry. The results showed that the distribution of Mg2+ of the arrhythmia cases was significantly lower than that of the control group (P < 0.01) and the concentration of Ca2+ in lymphocytes of arrhythmia case group was significantly higher than that of the control group (P < 0.01). The above distribution of Mg2+ and Ca2+ was somewhat related to the degree of heart failure. It is suggested that the lower distribution of Mg2+ in lymphocytes may cause arrhythmias. PMID- 10681807 TI - [A clinical analysis of 94 cases of recurrent stroke]. AB - Ninety-four cases of recurrent stroke were analyzed retrospectively, and 290 cases of first stroke episode were selected as controls. Results showed that recurrent stroke mainly appeared within the first year after the initial episode males predominant. The clinical manifestations of recurrent stroke were variable and commonly more severe. No significant differences was observed between the groups with respect to a variety of factors including the presence of hypertension, diabetes, history of transient ischemic attack, or familial history of stroke, cardiac attacks, cigarette smoking and/or alcohol consumption. The risk factors for recurrent stroke were discussed. PMID- 10681808 TI - [Effects of lotensin and nitrendipine on plasma fibrinogen and platelet aggregation in hypertensive patients]. AB - Plasma fibrinogen and platelet aggregation were measured by turbidimetric immunoassay, turbidimetry in 47 hypertensive patients and 20 normotensive control subjects. Among the 47 hypertensives, 24 cases were received lotensin and 23 nitrendipine. The plasma fibrinogen was increased and platelet aggregation enhanced in hypertensive patients before treatment. Platelet aggregation decreased after 8 weeks of treatment with lotensin or nitrendipine respectively. Lotensin decreased plasma fibrinogen whereas nitrendipine did not. It was concluded that both lotensin and nitrendipine decreased platelet aggregation; lotensin decreased plasma fibrinogen but nitrendipine did not. PMID- 10681809 TI - [Effects of lentinan of peripheral blood mononuclear cell expression of interleukin-2 receptor in patients with chronic hepatitis B in vivo and in vitro]. AB - The effects of lentinan on peripheral blood mononuclear cell (PBMC) expression of IL-2 receptor in patients with chronic hepatitis B in vitro and in vivo were studied. The expression percentage of IL-2 receptor in 44 patients with chronic hepatitis before treatment was 36.74 +/- 7.74, which was lower than healthy subjects, and was 48.92 +/- 3.79 after 2 months treatment of lentinan, which was approximated the normal control. PBMC from 30 patients with chronic hepatitis B were cultured in vitro with or without lentinan. The percentage of expression IL 2 receptor was 12.81 +/- 2.62 without lentinan and was 35.70 +/- 4.87 in the presence of lentinan. The concentration of lentinan stimulating expression of IL 2 receptor was between 0.2 to 200 g.ml-1. The results suggest that lentinan stimulates expression of IL-2 receptor on PBMC, which is associated with the therapeutic effectiveness of lentinan treatment. PMID- 10681810 TI - [Serum type IV collagen and laminin in patients with chronic hepatitis and its clinical significance]. AB - Serum levels of laminin and type IV collagen of 188 patients with different types of viral hepatitis were determined by RIA. Thirty-five blood donors were served as normal control. The results showed that: (1) higher levels of type IV collagen was found in patients with severe chronic hepatitis, post-hepatitis cirrhosis or subfulminant hepatitis; (2) the level of laminin was obviously increased in post hepatitis cirrhosis and subfulminant hepatitis; (3) there was a positive correlation between the levels of type IV collagen and laminin as well as gamma globulin, and negatively correlated to serum albumin, no correlation with alanine aminotransferase was observed; (4) detecting rate of hepatic fibrosis was increased with combination of serum laminin and type IV collagen examination; (5) serum type IV collagen was more sensitive than that of serum laminin. The results suggest that determination of serum type IV collagen and laminin are useful in diagnosis of hepatic fibrosis, and combination of the two parameters is recommended. PMID- 10681811 TI - [The clinic significance of serum hyaluronate and tumor necrosis factor alpha levels in liver diseases]. AB - Serum levels of hyaluronic acid (HA) and tumor necrosis factor alpha (TNF alpha) in 94 patients with various liver diseases and 31 healthy controls were studied by RIA, Serum TNF alpha in hepatitis, liver cirrhosis (LC) and primary hepatic carcinoma (PHC) was higher than that of the control. Serum HA in chronic hepatitis, LC and PHC was higher than that in the control. The serum level of HA showed a positive correlation with the serum level of TNF alpha. It is considered that the increase of serum HA indicates an early fibrogenic tendency in patients with liver diseases. TNF alpha is involved in the formation of hepatic fibrosis. PMID- 10681812 TI - [Domperidone and hyperprolactinemia]. AB - Nine patients with hyperprolactinemia and one patient with pituitary microadenona were observed for 6 years. They were induced by administration of domperidone. The clinical characteristic of the 9 patients was summarized and analysed. The results suggest that prescribe domperidone for fertile women should pay more attention to its adverse effect on inducing hyperprolactinemia and even pituitary microadenona. PMID- 10681813 TI - [The relationship between immunosuppression and oxygen free radicals in burned patients]. AB - The serum immunosuppression effect and its relation to oxygen free radicals were studied in patients with thermal injury (n = 11, total body surface area 20% 83%). The serum MDA, sIL-2R increased significantly in burned patients (P < 0.01 all). The serum of burned patients depressed the IL-2 productin of normal human PBMC (P < 0.01). Antioxidants decreased the serum sIL-2R in burned patients and attenuated the suppressive effect IL-2 production of normal human PBMC (P < 0.05 all). The results suggest that oxygen free radicals participate in the serum immunosuppression effect in burned patients. PMID- 10681814 TI - [A preliminary study of plasma oxidase activity by spectrophotometry on the healthy middle-aged and old volunteers in Changsha]. AB - The plasma oxidase activities (POA) in 26 healthy volunteers were determined by spectrophotometry. The results showed that the POA value of the 26 cases was obviously lower (78.90 +/- 8.12 U.L-1) than that in Lasla's (84 +/- 5 U.L-1) (P < 0.05). A preliminary normal reference range of POA (45-60 years old) in Changsha was established. This method is an accurate (CV = 3.9%), cheaper, and easy one and applicable for clinical laboratories. PMID- 10681815 TI - [Pathology and etiology of pulmonary alveolar proteinosis]. PMID- 10681816 TI - [Relation between plasma calcium and lipids in coronary disease]. PMID- 10681817 TI - [Relation between abnormal histogram and automatic blood platelet count]. PMID- 10681818 TI - [Skull damage caused by malignant changes of scalp cicatrix. A case report with radionuclide image]. PMID- 10681819 TI - [Application of microdissection, PCR, and microcloning technique on human chromosomal study]. PMID- 10681820 TI - [Study on relationship between apoptosis and proliferation of cells in liver cirrhosis and hepatocellular carcinoma]. AB - Apoptosis and nuclear antigen of proliferating cells were detected by labelling technique of in situ terminal deoxynucleotide transferase and immunohistochemical method in liver cirrhosis and hepatocellular carcinoma(HCC). The density of apoptotic cells in HCC was significantly lower than that in cirrhosis, and the density of proliferating cells was much higher in HCC than that in cirrhosis. Apoptotic cells mainly distributed in the peripseudolobular region of cirrhosis and formed an apoptosis zone. But they scattered within the cancer tissue. The results suggest that the formation of apoptosis zone in cirrhosis may be related to the change of liver blood stream. Selective proliferation of cells may exist during carcinogenesis of liver cirrhosis. PMID- 10681821 TI - [The effect of brain-derived neurotrophic factor on the rat electroretinography after pressure-induced ischemic injury]. AB - Brain-derived neurotrophic factor (BDNF) or normal goat serum (NGS) was injected intravitreously in each 11 wistar albino rats two days before the induction of ischemia. Retinal ischemia was induced in 22 rats by increasing intraocular pressure to the threshold level which extinguished the electroretinography (ERG) b-wave individually, and maintaining for 90 minutes. The ERG b-wave of the BDNF treated rats recovered to 80.5 +/- 24.4% of their normal amplitude three days after ischemia, while that of the NGS-treated rats recovered only to 11.1 +/- 5.3%. Statistic analysis showed that the recovery level of ERG b-wave after ischemia in the BDNF and NGS treated rats was significantly different (P < 0.01). The results suggest that BDNF could promote the recovery of the retinal electrophysiologic function from ischemia induced by high intraocular pressure. PMID- 10681822 TI - [Drug resistance study and detection of methicillin-resistant staphylococci]. AB - MRSA and MRSE were identified by detecting oxacillin resistance of 107 strains of staphylococci from clinical sources. The positive rates of MRSA were 45.2% and 16.9%, respectively among the whole number of staphylococci examined. Antibiotic resistant strains were detected with disks by Kirby-Bauer method. All MRSA and MRSE strains were multi-resistant. Resistant rates of MRSA and MRSE were higher than MSSA and MSSE. The positive rates of beta lactamase producing in MRSA and MRSE were significantly higher than MSSA and MSSE (P< 0.05 and P< 0.01) respectively. These results indicated that MRSA and MRSE were the main pathogens in the clinical infection. MRSA and MRSE detection are important for the early diagnosis and treatment of patients with infective diseases. PMID- 10681823 TI - [Expression and significance of p53, c-erbB2, CEA proteins in colorectal adenoma and carcinoma]. AB - Expression of p53, c-erB2, CEA proteins in 15 cases of normal colorectal mucosa, 40 adenoma and 40 carcinoma of colon were studied with immunohistochemistry. The positive rate of p53, c-erbB2, CEA in colorectal carcinomas were significantly higher than those in adenoma. The positive rate of p53 in adenoma with severe atypical hyperplasia was higher than that in the mild (P< 0.01), and was higher in colorectal carcinomas of poor differentiation that that in the well differentiated ones (P< 0.05). The novel type monoclonal antibody of CEA Col-1 had a high sensitivity and specificity for the tissue of colorectal carcinoma in which the positive rate was 85%. The results suggested the alteration p53, c erbB2 are involve in the tumorgenesis and development of colorectal carcinoma; and detection of p53, c-erbB2 and CEA proteins are helpful in distinguishing benign and malignant polyps of colon. PMID- 10681824 TI - [Study on the effects of hyaluronic acid-streptomycin perfusion through the round window on the function and morphology in guinea pig inner ears]. AB - To investigate the effects of the hyaluronic acid-streptomycin (HA-SM) perfusion through round window on the function and morphology of the inner ear in guinea pig, membrous labyrinth mapping, temporal bone section after celloidin embedding, transmission electron microscopy, electrocochlegraphy (ECochG) and electronystagmography (ENG) were examined. The nystagmus duration induced by caloric test was obviously reduced in comparison with that of the preoperation (P< 0.01), while the action potential(AP) by ECochG was not obviously changed. The sensory cells of estibular organs were severely damaged, while the morphology of corti's organs were significantly damaged after HA-SM perfusion. The results suggest that the HA-SM perfusion through the round window may selectively destroy the vestibular function, whereas the auditory function is not obviously damaged. PMID- 10681825 TI - [Effects of dibutyl phthalate on the proliferation and apoptosis of leukemic cells]. AB - This study was designed to investigate the effects of dibutyl phthalate (DBP) on the proliferation and apoptosis of leukemic cells. The results showed that DBP suppressed the proliferation of HL-60 and K562 leukemic cells in a dose-dependent and time-dependent manner. Inducing HL-60 and K562 leukemic cells to die via apoptosis were confirmed by that the percentage of the apoptotic cells in morphology and of the DNA fragmentation increased significantly and that typical ladders of apoptosis were shown by gel electrophoresis when HL-60 and K562 cells were exposed to the DBP. Also, K562 leukemic cells was less susceptible to the DBP than HL-60 cells. These results indicate that DBP can inhibit the proliferation of HL-60 and K562 leukemic cells and induced them to die via apoptosis. PMID- 10681826 TI - [A comparative study on the risk factors for the incidence of tongue cancer]. AB - The risk factors for the incidence of tongue cancer were analyzed with case study. The result showed that oral leukoplakia (OR = 8.50) residual roots of oral teeth (OR = 8.50), eating hot foods (OR = 13.0) and eating fire-roasted foods (OR = 3.5) were the main risk factors for the occurrence of tongue cancer. Eating fresh fruits (OR = 0.01) was usually helpful to the decrease in the predisposition of tongue cancer. It suggested that oral precancerous lesions and mouth diseases should be cured early and good life behavior must be developed in order to prevent the incidence of tongue cancer. PMID- 10681827 TI - [Effect of dietary intake on body mass index in middle aged and elderly population]. AB - The paper investigated and analyzed the relationship between the nutrient intake and body mass index (BMI) of 200 middle-aged and elderly people(aged 35 - 74 years) in Changsha. The result showed that the constituent rate of underweight (BMI< 18.5) was 6.5% and that of over-weight (BMI<25.0) was 23.5%. The amount of fat intake constituted slightly higher proportion in energy intake. The stepwise regression analysis indicates that the body mass index increases with the increasing intake of carbohydrates. PMID- 10681828 TI - [Open heart surgery without cross clamping aorta]. AB - Open heart surgery without cross clamping aorta under normothermic or hypothermic cardiopulmonary bypass was performed in 167 patients in which there were more than ten kinds of cardiac diseases. The characteristics of the procedure included only vena cava blocked and aorta unclamped with beating heart or artificial evoked ventricular fibrillation. It diminished myocardial ischemia and reperfusion injuries maximally. There were no low cardiac output syndrome, severe arrhythmia and complications of brain except two patients died postoperatively. The operative mortality rate was 1.1% (2/167). The advantages, disadvantages and safety of this method are discussed briefly in this paper. PMID- 10681829 TI - [Effect of apolipoprotein E gene plasma levels of lipids, lipoproteins, apolipoproteins and its relation with coronary heart disease]. AB - Using a technique of polymerase chain reaction-restriction lengths polymorphism (PCR-RLP), the authors detected the genotypes of apolipoprotein EE(ApoE) in 102 normal control subjects and in 118 patients with coronary heart disease (CHD). Meanwhile, the effect of ApoE allele on plasma lipid, lipoprotein, apolipoprotein levels were analyzed, and ApoE allele frequencies of patients with CHD and of control subjects were compared. The results showed that the technique of modified PCR-RFLP was a useful measure to detect the genotypes of ApoE. ApoE gene was polymorphism, resulting in plasma lipid level variance among individuals. As compared to ApoE3 allele, ApoE4 allele had positive effect on plasma TC, LDL-C and ApoB100 levels; while ApoE2 allele had negative effect on plasma TC, LDL-C and ApoB100 levels. The frequency of E4 allele in patients with CHD was higher than that in control subjects. The results in the present study suggest that ApoE4 allele is probably the genetic marker of CHD. PMID- 10681830 TI - [Influence of cardiopulmonary bypass time on gastric intramucosal Pco2 and pH]. AB - To determine the influence of cardiopulmonary bypass time on gastric intramucosal Pco2 and pH (pHi), 16 patients undergoing cardiopulmonary bypass (CPB) were divided into two groups according to the CPB time-- group A (n = 8) within 100 minutes and group B (n = 8) beyond 100 minutes. The gastric intramucosal Pco2 increased and the pHi decreased significantly (P< 0.01) at the end CPB compared with before operation, before bypass and 30 minutes after intermission of CPB in all patients. Between group B and A, there were significant differences in the gastric intramucosal Pco2 and in the pHi at the end of CPB. The patients of group A had no severe complications. In group B, three patients developed life threatening complications, one of them died. The results indicate that the longer the CPB time, the greater the degree of gastric mucosal acidosis. It predicts the patients may develop life-threatening complication after cardiac operations. PMID- 10681831 TI - [Study on dopamine metabolite in cerebrospinal fluid of schizophrenics and epilepsics]. AB - The concentration of dopamine (DA) metabolite-homovanillic acid (HVA) in cerebrospinal fluid (CSF) of 40 patients with schizophrenia 27 with epilepsy and 15 controls without disorders of CNS was reported. The results showed that there were no significant differences in mean levels of HVA among the three groups. It is suggested that both schizophrenia and epilepsy are not abnormal in total level of DA metabolism. PMID- 10681832 TI - [A study on the activity of nitric oxide in alveolar macrophages from patients with lung cancer]. AB - Nitrite and nitrate (NO2-/NO2-) in the bronchus alveolar lavage fluid (BALF) and the supernatants of incubated alveolar macrophages (AMs) from patients with primary lung cancer were measured by copper-coated cadmium reduction and Griess method. Mrna expression of AM induced nitric oxide synthase (iNOS) were analyzed by RT-PCR. There was NO2-/NO2- in BALF either from lung cancer patients or from control subjects. When compared with control group and the nontumor-bearing lung, the level of NO2-/NO2-was lower in BALF from the tumor-bearing lung [5.18+/-1.1 vs 2.47+/-0.67nmol x mg protein-1 (P< 0.01); 4.65+/- 2.46 vs 2.47+/- 0.67nmol x mg protein-1(P< 0.01)]. We also found a lower level of NO2-/NO2- in the supernatants of incubated AMs from the lung of cancer patients than from control and nontumor-bearing lung [95.03+/- 21.76 vs 63.37+/- 17.58nmol (P< 0.01); 85.61+/- 16.70 vs 63.37+/- 17.58nmol (P< 0.05)]. No significant difference existed between the MRNA expression of AM iNOS in lung cancer patients (69%) and that of control subjects (91%). After the AMs were stimulated with granulocyte macrophage colony stimulating factor (GM-CSF), the level of NO2-/NO2- in the supernatants was significantly increased (P< 0.01); while the mRNA expression of AM iNOS from patients with lung cancer resulted in an increase of 16.85+/- 7.58% vs 33.38+/- 8.21% of control group (P< 0.05). These observation suggest that some defects of antitumor function occur in the AMs at the tumor region. GM-CSF can stimulate AMs and thus potentiate their NO activity. PMID- 10681833 TI - [Gamma knife treatment of meningiomas]. AB - From October 1995 to July 1997, 52 cases of meningioma were treated with gamma knife radiosurgery. Twenty-eight cases were followed up by CT or MR. The follow up period was from 2 to 22 months. Follow-up imaging evaluation showed tumor size shrinkage in 8 cases, central density decrease in 10 cases, no change in 8 cases and tumor volume increase in 2 cases. The actual 1-year tumor growth control rate was 92%. Radiation-induced edema was noted in 9 cases, including 7 cases of the nonbasal tumor and 2 cases of basal one. The indications and efficacy of gamma knife treatment of the meningioma were discussed. Our preliminary experience suggests that gamma knife is an effective and safe technique for carefully selected patients with meningiomas. PMID- 10681834 TI - [Clinical significance of pulmonary valvular resistance in children determined by color Doppler echocardiography]. AB - Pulmonary valvular resistance (PVR) of 40 children with pulmonary valvular stenosis was determined by color Doppler echocardiography before and after percutaneous balloon pulmonary valvuloplasty as well as during their follow-up. The correlation coefficient of PVR between Doppler method and catheterization method was 0.85 ( P <0.01) and the correlation coefficient between PVR determined by former method and pulmonary valvular area determined by the latter was -0.80 ( P <0.01). This study indicates that color Doppler echocardiography can take the place of catheterization to determine PVR in assessing the severity of pulmonary valvular stenosis in children. PMID- 10681835 TI - [Bilobate flap applied to the treatment of secondary deformity after cheiloplasty]. AB - Secondary deformity after cheiloplasty was common and 7 cases (4 males, 3 females) were reconstructed with bilobate flap. The patients' age ranged from 16 to 30. Five of them with single cleftlip. The deformity of them included loose and shortening of the upperlip, scar, vermilion pitting, too large nostril etc. Lateral tissue of cleftlip was used to construct the bilobate flap which was applied to repairing secondary deformity after cheiloplasty. The method was matched with the principle of anatomy, and the effects were satisfactory. PMID- 10681836 TI - [Infection of sexually transmitted diseases in female patients with inflammation of genitourinary tract by polymerase chain reaction]. AB - Using polymerase chain reaction (PCR), Neisseria gonorrhoeae (NG) from 5899 female out-patients with inflammation of genitourinary tract was detected. Among 18.57% positive cases, the effective rates of the gonorrhea (26.07%) and urethritis (23.73%) were higher than those of the Condyloma acuminatum (CA) (17.73%), vaginitis (17.70%), and cervicitis (11.54%)(P< 0.01), indicating that there were various degrees of NG infection in genitourinary tract diseases. Moreover, Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), NG, and human papilloma viruses (HPV) were also detected in 8329 patients with vaginitis. Among 20.01% positive cases, the positive rate of UU was 39.95%, CT 21.36%, NG 17.70%, and HPV 16.40%. Variance analysis showed that significant differences existed in the vaginitis caused by different pathogens ( P <0.01); namely, four kinds of pathogens were present in the patients with vaginitis. PMID- 10681837 TI - [Clinical analysis of early complications in 132 low body weight infants performed open-heart surgery]. AB - A clinical analysis of early complications in 132 low body weight infants undergoing open-heart surgery is reported. There were a higher risk of early complication associated with severe illness and/or experienced a longer duration time of cardiopulmonary bypass and endotracheal tube. The total complication rate was 42.4% and the major three complications were low cardiac output, acute respiratory distress syndrome and problems from endotracheal tube (occlusion and/or in proper position of tube). The author emphasized the importance of management of endotracheal tube after operation. PMID- 10681838 TI - [CT diagnosis of carcinomas of ovary]. AB - OBJECTIVES: To evaluate CT imaging feature and CT in staging of the ovarian carcinomas and low malignant potential tumors (LMP) in relation with pathology. METHODS: Preoperative CT image of of 48 tumors, including 45 carcinomas and 3 LMP in 30 patients verified surgical-pathologically were analyzed. RESULTS: The tumors were classified as cystic, predominantly cystic, mixed, predominantly solid and solid types. The solid and solid portion of the mass were enhanced obviously. It appeared flower-shaped or nodular shadows or irregularity of patch. Cystic portion of the mass with a wall or septa thickness exceeding 3mm observed wall nodularily and/or a solid mass. LMP appeared purely cystic or predominantly cystic, accompanied with a thin regular wall, thin septa and regular solid portion with contrast material uptake. CT permitted accurate estimation of the FiGo in 80% of patients. CONCLUSION: CT images is helpful to explore the pathologic appearances of neoplasms and biological characteristics of ovary carcinoma. PMID- 10681839 TI - [The rescue of iodine allergy by contrast enhanced CT]. AB - We summarized 51 cases of iodine allergy and allergic shock induced by contract enhanced computed tomography (CT). It is suggested that (1) the appropriate measures must be taken to prevent the side effects of ionic contrast medium; (2) a set of first-aid drug and essential equipment must be prepared; (3) the staff of CT room must be trained with the first-aid techniques and knowledge. PMID- 10681840 TI - [The value of color Doppler ultrasonography in diagnosing aneurysm]. AB - Using Doppler ultrasound, we detected 59 patients with aneurysm, 54 cases were confirmed by angiography or operation or both. Among them, 33 cases (63%) showed true aneurysm, 18 cases (33%) pseudoaneurysm, and 2 cases (4%) superficial congenital arterio-venous aneurysm. Eight cases were associated with dissected stripping. One case was falsely diagnosed by ultrasonography. The diagnostic accuracy was 98%. The results suggest that color Doppler ultrasonography is useful in detecting aneurysm. PMID- 10681841 TI - [Effects of monocyte-derived endothelin on acute cerebral infarction]. AB - To investigate the mechanism that monocytes participate in ischemic brain injury, endothelin (ET) levels in plasma, supernatants of cultured monocytes in vitro and those pretreated by dexamethason (DXM) were assayed by radioimmunoassay in 31 patients with acute cerebral infarction and 16 patients with hypertension (served as control). Compared with the control group, the ET levels in supernatants of cultured monocytes increased and positively related with those in plasma and degree of neurological damage in the acute cerebral infarction group, while the ET levels in supernatants of cultured monocytes pretreated by DXM decreased. This suggests that intensifying the synthesis and release ET may be one of the mechanisms for the involvement of monocytes in the pathologic process of ischemic brain injury during acute cerebral infarction. DXM can improve this process by inhibition of producing monocyte-derived ET. PMID- 10681842 TI - [Clinical of 50 cases of aged patients with pneumonia caused by gram-negative bacteria]. AB - The diagnosis and treatment of 50 cases of aged patients with pneumonia caused by gram-negative G-bacteria (from Feb. 1993 to Aug. 1997) were analyzed. Before the results of drug sensitive tests were reported, aminoglycosides plus broad spectrum semisynthetic penicillin or first generation cephalosporin could improve the curative effect in patients with good renal function. Bacterial culture and drug sensitive tests were crucial for choosing the proper antibiotics. PMID- 10681843 TI - [Surgical treatment of sacral tumor]. AB - From 1990 to 1996, 21 patients with sacral tumor were surgically, including 8 cases with giant tumor of bone, 7 cases of spinal cord tumor, each 2 cases of neurofibroma and adenoma, 1 case of myeloma and 1 case of lipoma with low grade of malignancy. A total of 22 operations involving one for recurrent tumors in 21 cases were performed. Sacral resection and curettage plus resection were the surgical ways. 19 patients were followed up with an average period of 2.5 years. 15 patients showed good results, 3 patients occurred urinary incontinence and constipation, one of 3 cases occurred weakness of ankles and feet. Authors conclude that surgery should be advised and actively adopted for sacral tumors. PMID- 10681844 TI - [Detection of interleukin-2 receptor in patients with idiopathic thrombocytopenic purpura]. AB - Using ELISA and indirect fluorescent antibody techniques(IFAT), the serum soluble inter-leukin-2 receptor(sIL-2R) and membrane IL-2R (mIL-2R) on lymphocyte cells induced by phyto-hemagglutinin (PHA) were studied in patients with idiopathic thrombocytopenic purpura (ITP). The results showed that the level of sIL-2R increased significantly before treatment and decreased after treatment; the percentages of mIL-2R induced by PHA decreased. There was no correlation between the level of serum sIL-2R and that of mIL-2R. Our observations suggest that increased level of serum sIL-2R in ITP patients may be one of causes evoking cellular immunological abnormalities. PMID- 10681845 TI - [MRI observation on Arnold-Chiari I malformation]. AB - Craniocerical junction of 42 patients with Arnold-Chiari I malformation (ACM-1) and 41 normal controls were studied. In addition to the decendent tonsil, all the components in the posterior cranial fossa including the 4th ventricle, oblongata medulla and the sinus confluence of patients with ACM- 1 were decended. Although the spinal foramen magnum anterior -posterior diameter was smaller but not statistically different from the control. The results suggest that the main pathology of ACM- 1 is the shallow and small posterior cranial fossa and the resulted organ decendent as a whole. PMID- 10681846 TI - [Antiviral and antibacterial actions of bingduqing granules in vitro]. AB - The antiviral and antibacterial actions in vitro of bingduqing granules which consists of nine kinds of Chinese traditional medicinal herbs were observed. The results showed that the inhibitive concentrations of bingduqing granules for Adv 7, RSV, HSV-1, and influenza virus A3, were >-6.9, 8.3, 13.8 and 83 mg x ml-1, respectively; while the minimum inhibitive concentrations for Staphylococcus aureus, Staphylococcus epidermidis, Type A and B Streptococci, and pseudomonas aeruginosa were 0.03125, 0.03125, 0.125, 0.125, and 0.25 g x ml-1, respectively. This study provide a pharmacodynamics basis for clinical application of Bingduqing Granules in the treatment of respiratory tract infection. PMID- 10681847 TI - [Culture of murine HPP-CFC in vitro using conditioned medium substitutefor recombinant growth factors]. AB - OBJECTS: To study the clonal growth on single layer, agar of high proliferative potential colony-forming cells (HPP-CFC) of bone marrow obtained from both normal mice (NBM) and mice treated 2 days earlier with 5-fluorouracil (FU,BM) using conditioned media. MATERIAL AND METHODS: Using liquid cell culture to harvert WEHI2-CM and L929-CM as the substitutes for IL-3 and M-CSF respectively; using semi-solid culture to assay HP-CFCin vitro. RESULTS: (1) the combination of WEHI3 CM and L929-CM produced HPP-CFC in vitro, and the addition of rhIL-1 alpha, rhIL 6 and rmGM-CSF further enhanced HPP-CFC was enriched in the FU2 BM. Morphological investigation showed that the HPP-CFC colony was large and compact, great than 0.5mm in diameter and contained more than 50,000 cells, most of which were macrophage like cells. The hydroxyurea suicide rate for HPP-CFC was less than 10%. CONCLUSION: The results suggested that (1) WEHI3-CM and L929-CM can be used as the substitutes of IL-3 and M-CSF to stimulate the formation of HPP-CFC; (2) FU can enrich HPP-CFC of the mice. PMID- 10681848 TI - [Study on the clinical diagnostic values of MCV and RDW to homolytic anemia]. AB - In this study, the mean corpuscula volume (MCV) and the red blood cell distribution width (RDW) were measured in 21 patients with hemolytic anemia, 35 patients with non-hemolytic anemia, and 100 healthy subjects were in the control group. The result revealed that the changes of MCV and RDW in hemolytic anemia and the other proliferative anemia were special. As diagnostic criterion of HA, MCV and RDW showed high-sensitivity and specificity. It is useful for screening HA patients. PMID- 10681849 TI - [The factors affecting apoptosis by in situ terminal deoxynucleotide transferase method]. AB - Apoptosis in biopsies of patients with hepatocirrhosis or hepatocellular carcinoma and in small intestinal mucosa of mice were detected by the method of in situ terminal deoxynucleotide transferase (ISTdT). It was found that the best positive result might be obtained by digesting tissue sections with 25mg x L-1 proteinase K for 15 minutes, then treating sections with moderate degree of microwave for 5 minutes after reaction with terminal deoxynucleotide transferase. The best choice of above conditions were discussed. PMID- 10681850 TI - [Boari flap ureteroplasty in the treatment of distal long-segment defect or stricture of the ureter]. PMID- 10681851 TI - [Human sex identification in dried bloodstains by PCR using X-Y homologous primers]. PMID- 10681852 TI - [Cerebrospinal fluid cytology in the diagnosis of cancerous meningitis]. PMID- 10681853 TI - [Clinical analysis of acute glomerulonephritis in 166 children]. PMID- 10681854 TI - [Effects of aprotinin on activated coagulation time in the patients during cardiopulmonary bypass]. PMID- 10681855 TI - [Relation between fibromyalgia and bacterial urine]. PMID- 10681856 TI - [A case of multiple pathway associated with right bundle branch block]. PMID- 10681857 TI - [Magnesium sulfate in the treatment of point torsion of ventricular tachycardia and fibrillation caused by haotenggen. A case report]. PMID- 10681858 TI - [Large calcified cerebral glioma. Report of 2 cases]. PMID- 10681859 TI - [Right ventricular myocardial infarction with cardiac aneurysm. A case report]. PMID- 10681860 TI - Literature research on screening of the nucleus acupoints for treatment of intellectual disturbances. PMID- 10681861 TI - Clinical analysis on the treatment of HIV/AIDS by traditional Chinese medicine. PMID- 10681862 TI - Treatment of atrophic cholecystitis by regulating the spleen--a report of 50 cases. AB - Fifty cases of atrophic cholecystitis were treated by regulating of the spleen. Of them, 21 were cured, 18 remarkably effective, and 7 effective. The overall effective rate was 92.0%. As compared with the results of ultrasonography B performed before and after treatment, it was shown that both the longitudinal and transverse inner diameters of the gallbladder increased evidently, and the condition of atrophy improved remarkably after treatment. PMID- 10681863 TI - An analysis for death causes in 45 cases of liver cancer treated with traditional Chinese drugs. AB - Among the 165 cases of late-stage liver cancer treated in our hospital, 65 (39.4%) died, with an average survival time of 8.1 months and a median survival time of 7 months. Among the 65 dead patients, 45 were treated with traditional Chinese drugs and 20 with western medicine. The average survival time was 8.4 months in the former and 7.3 months in the latter group. The direct causes of death for the 65 patients were hepatic coma, severe hemorrhage of the upper digestive tract, Heyd's syndrome, hepatorrhexis, respiratory failure, cardiac failure, etc. The incidence rates of hemorrhage of the upper digestive tract and hepatorrhexis in the 45 patients treated with traditional Chinese drugs were obviously lower than those treated with western medicine. PMID- 10681864 TI - 311 cases of chronic osteomyelitis treated by soaking with ganlingsan liquid. AB - From 1992 to 1995, 311 cases of chronic osteomyelitis (including 145 cases of hematogenic osteomyelitis, 147 cases of traumatic osteomyelitis, 8 cases of secondary osteomyelitis after local infection, 4 cases due to other causes, and 7 cases with cause unknown) were treated by soaking with Ganlingsan liquid. The average treatment course was 69.5 days. The results showed that 260 cases were cured, 26 cases markedly effective, 20 cases improved, and 5 cases ineffective. The evaluation of the function revealed that 150 cases were excellent, 129 cases good, 10 cases improved, and 22 cases poor. 111 patients recovered from abnormal erythrocyte sedimentation rate. External fixation was applied at the same time for the patients accompanied by nonunion of fracture, and the therapeutic effect was also satisfactory. PMID- 10681865 TI - 380 cases of bronchiectasis with hemoptysis treated by point-injection. PMID- 10681866 TI - Treatment of epiphora due to insufficiency of lacrimal passage by acupuncture at Jingming. AB - Since 1995, the authors have employed acupuncture at Jingming (UB 1) with the warming needles to treat epiphora due to insufficiency of lacrimal passages. Of the 68 treated eyes in 42 patients, 28 eyes were cured, 35 improved, and 5 ineffective with a total effective rate of 92.65%; and 12 eyes were cured and 34 improved by only one course of treatment. PMID- 10681867 TI - Observation of the efficacy of acupuncture and moxibustion in 62 cases of chronic colitis. AB - 62 patients with chronic colitis were randomly divided into two groups. Acupuncture and moxibustion at acupoints such as Tianshu (St 25), Guanyuan (Ren 4) were applied in the treatment group, and western drugs were applied in the control group. The results showed that acupuncture and moxibustion had a marked curative effective with few side effects, and therefore was readily acceptable to the patients. PMID- 10681868 TI - Twenty-three cases of atrophic rhinitis treated by deep puncture at three points in the nasal region. AB - Atrophic rhinitis is a disease which manifests itself mainly by anosmia due to dryness and atrophy of the nasal mucosa. There is no specific therapy for the disease at present. In the past few years, 23 cases of atrophic rhinitis were treated mainly by deep puncture at three points in the nasal region with satisfactory results. In order to find out the functional changes of the nasal mucosa, the mucociliary transport rate (MTR), surface temperature of the conchal mucosa, acid-base scale of nasal secretion, and volume of nasal secretion were determined before and after the treatment. PMID- 10681869 TI - Dr. Du Xiaoshan's personal experience in acupuncture treatment. PMID- 10681870 TI - A parallel study on the effects in treatment of impotence by tonifying the kidney with and that without improving blood circulation. AB - 141 cases of functional impotence of the kidney-deficiency type were treated by tonifying the kidney. On them, 103 cases at the same time were treated by improving blood circulation and the other 38 cases by the former only. As a result the total effective rate and the markedly effective rate in the former were 84.46% and 46.60% respectively; but in the latter, 60.55% and 13.15%. A significant difference was found in Ridit analysis (P < 0.05), indicating that method of tonifying the kidney with improving blood circulation is much better than by simply tonifying the kidney alone. PMID- 10681871 TI - Clinical application of ciliao in acupuncture treatment. PMID- 10681872 TI - A clinical observation on therapeutic effects of acupuncture for allergic rhinitis. PMID- 10681873 TI - Study on pathogenic mechanism of emotions in traditional Chinese medicine--an observation of hydrogen peroxide releasing function of celiac macrophages in rats under stress state. AB - The hydrogen peroxide releasing function of macrophages in rats under the stress state was observed with the animal model of "excessive anger impairing the liver". The results showed that the volume of hydrogen peroxide released from the macrophages in rats was decreased, while the corticosterone level in plasma increased after stress. It indicates that stimulation of harmful emotions could cause inhibition of immunoreaction of the organism, which might be related to the enhancement in excitability of the hypothalamus-pituitary-adrenal axis and hypersecretion of glucocorticoid hormone. PMID- 10681874 TI - Effects of transient forebrain ischemia and radix Salviae miltiorrhizae (RSM) on extracellular levels of monoamine neurotransmitters and metabolites in the gerbil striatum--an in vivo microdialysis study. AB - The aim of this study was to investigate the effect of 30 min forebrain ischemia, followed by 120 min reperfusion on extracellular fluid (ECF) levels of dopamine (DA), norepinephrine (NE), serotonin (5-HT) and their metabolites, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) in the striatum of gerbils, so as to obtain further information on the mechanism of Radix Salviae Miltiorrhizae (RSM)-induced neuroprotection. Microdialysis was used to sample the extracellular space. Dialysate was measured by high performance liquid chromatography with electrochemical detector (HPLC-ED). ECF DA, NE levels increased from basal levels by 282, 227 and 221 folds, by 9.14, 8.51 and 8.25 folds, respectively for the three ischemic duration (0-10; 11-20; 21-30 min). ECF DA, NE, 5-HT levels in the RSM-treated group were significantly decreased as compared with those in the control group during ischemia (P < 0.01). The results suggested that monoamine neurotransmitters were involved in ischemic neuron damage directly or indirectly; and that RSM plays a protective role during cerebral ischemia by attenuating the dysfunctions of monoamine neurotransmitters. PMID- 10681875 TI - Reciprocal actions of acupoints on gastrointestinal peristalsis during electroacupuncture in mice. AB - Orthogonal design was used to observe the gastrointestinal peristalsis in normal and atropine-treated mice after electroacupuncture was applied, singly or in combination, at Neiguan (P 6), Pishu (UB 20) and Zusanli (St 36). The results showed that: 1) electroacupuncture has no significant effect on the gastrointestinal peristalsis in normal mice; 2) Pishu (UB 20) was significantly antagonistic to Zusanli (St 36) in normal mice; 3) the decreased gastrointestinal peristalsis in atropine-treated mice was markedly promoted by electroacupuncture at Zusanli (St 36); and 4) Neiguan (P 6) was significantly antagonistic to Pishu (UB 20) in atropine-treated mice. The results indicated that the reciprocal actions among acupoints should be taken into consideration for point prescription. PMID- 10681876 TI - Considerations in making prescriptions for diabetes. PMID- 10681877 TI - Clinical application of radix Aconiti lateralis preparata. PMID- 10681878 TI - Rationale for the designing of a new model of compound electroacupuncture moxibustion stimulator. PMID- 10681879 TI - International Anesthesia Research Society 74th Clinical and Scientific Congress. Honolulu, Hawaii, USA. March 10-14, 2000. Abstracts. PMID- 10681880 TI - Abstracts of meetings of the International Federation of Clinical Neurophysiology, 1998-1999. PMID- 10681881 TI - Pathological Society of Great Britain and Ireland 178th meeting. 6-8 January 1999. Abstracts. PMID- 10681882 TI - Pathological Society of Great Britain and Ireland 179th meeting. 7-9 July 1999. Abstracts. PMID- 10681883 TI - XXVII Symposium of the Italian Cancer Society. Milan, November 22-24, 1999. Abstracts. PMID- 10681884 TI - Socioeconomic status and health: what we know and what we don't. AB - In the past 15 years, we have seen a marked increase in research on socioeconomic status (SES) and health. Research in the first part of this era examined the nature of the relationship of SES and health, revealing a graded association; SES is important to health not only for those in poverty, but at all levels of SES. On average, the more advantaged individuals are, the better their health. In this paper we examine the data regarding the SES-health gradient, addressing causal direction, generalizability across populations and diseases, and associations with health for different indicators of SES. In the most recent era, researchers are increasingly exploring the mechanisms by which SES exerts an influence on health. There are multiple pathways by which SES determines health; a comprehensive analysis must include macroeconomic contexts and social factors as well as more immediate social environments, individual psychological and behavioral factors, and biological predispositions and processes. PMID- 10681885 TI - Epidemiology of socioeconomic status and health: are determinants within countries the same as between countries? AB - Within societies, health and ill-health follow a social gradient: lower socioeconomic position, worse health. The slope of the gradient has varied over time. It is likely that social and economic circumstances play a role in this changing slope. Within Europe, differences in health between East and West have also varied in magnitude. The advantage in the West increased through the 1970s and 1980s. Although similar in their health trends up to 1989, the countries of central and eastern Europe have diverged quite sharply subsequently. It is again likely that changing social and economic fortunes account for these trends. There is evidence to support the role of psychosocial factors in relating to socioeconomic differences within and between countries. PMID- 10681886 TI - Protective and damaging effects of mediators of stress. Elaborating and testing the concepts of allostasis and allostatic load. AB - Stress is a condition of human existence and a factor in the expression of disease. A broader view of stress is that it is not just the dramatic stressful events that exact their toll but rather the many events of daily life that elevate activities of physiological systems to cause some measure of wear and tear. We call this wear and tear "allostatic load," and it reflects not only the impact of life experiences but also of genetic load; individual habits reflecting items such as diet, exercise, and substance abuse; and developmental experiences that set life-long patterns of behavior and physiological reactivity (see McEwen). Hormones associated with stress and allostatic load protect the body in the short run and promote adaptation, but in the long run allostatic load causes changes in the body that lead to disease. This will be illustrated for the immune system and brain. Among the most potent of stressors are those arising from competitive interactions between animals of the same species, leading to the formation of dominance hierarchies. Psychosocial stress of this type not only impairs cognitive function of lower ranking animals, but it can also promote disease (e.g. atherosclerosis) among those vying for the dominant position. Social ordering in human society is also associated with gradients of disease, with an increasing frequency of mortality and morbidity as one descends the scale of socioeconomic status that reflects both income and education. Although the causes of these gradients of health are very complex, they are likely to reflect, with increasing frequency at the lower end of the scale, the cumulative burden of coping with limited resources and negative life events and the allostatic load that this burden places on the physiological systems involved in coping and adaptation. PMID- 10681887 TI - Health, hierarchy, and social anxiety. AB - This paper suggests that the main reasons why populations with narrower income differences tend to have lower mortality rates are to be found in the psychosocial impact of low social status. There is now substantial evidence showing that where income differences are greater, violence tends to be more common, people are less likely to trust each other, and social relations are less cohesive. The growing impression that social cohesion is beneficial to health may be less a reflection of its direct effects than of its role as a marker for the underlying psychological pain of low social status. Low social status affects patterns of violence, disrespect, shame, poor social relations, and depression. In its implications for feelings of inferiority and insecurity, it interacts with other powerful health variables such as poor emotional attachment in early childhood and patterns of friendship and social support. Causal pathways are likely to center on the influence that the quality of social relations has on neuroendocrine pathways. PMID- 10681888 TI - Developmental influences across the life span. PMID- 10681889 TI - Maternal care, gene expression, and the development of individual differences in stress reactivity. PMID- 10681890 TI - The biological embedding of early experience and its effects on health in adulthood. AB - Explanations of the socioeconomic gradient in health status must account for the observations that the gradient cuts across a wide range of disease processes and is capable of replicating itself on new disease processes as they emerge in society. Understanding this pattern requires an understanding of how human organisms can become generally vulnerable or resilient to disease over time: a huge collation task across different disciplines. The hypothesis that best fits current evidence is that the gradient is an "emergent property" of the interaction between the developmental status of people and the material and psychosocial conditions they encounter over their life course. Within this broad formulation, special attention is given to child development, and the prospect that socioeconomic differences in the quality of early life experiences contribute to subsequent gradients in health status through socioeconomic differences in brain sculpting and the conditioning of host defense systems that depend on communication with the developing brain. The contribution to the gradient in health is theorized to occur through a combination of latent effects, pathway effects, and cumulative disadvantage. PMID- 10681891 TI - Hierarchies of life histories and associated health risks. AB - Widely documented inverse associations between socioeconomic standing and incident chronic disease and mortality invite explanation in terms of pathways to these outcomes. Empirical identification of pathways, or histories, requires measures that assess cumulative wear and tear on physiological systems following from psychosocial adversity and genetic predispositions. Such an assessment, allostatic load, has been shown to predict later life mortality, incident cardiovascular disease, and decline in physical and cognitive functioning. Using data from the Wisconsin Longitudinal Study (WLS), we seek precursors to allostatic load via ordered categories of cumulative adversity relative to advantage over the life course. We operationalize these histories via unfolding economic circumstances and social relationship experiences (e.g., parent-child interactions, quality of spousal ties). Findings reveal a strong direct association between the extent of adversity relative to advantage in an ordering of these histories and likelihood of high allostatic load. Importantly, resilient individuals with economic disadvantage, but compensating positive social relationship histories also show low prevalence of high allostatic load. PMID- 10681892 TI - What is the role of the social environment in understanding inequalities in health? PMID- 10681893 TI - Social capital and community effects on population and individual health. AB - Social capital refers to those features of social relationships--such as levels of interpersonal trust and norms of reciprocity and mutual aid--that facilitate collective action for mutual benefit. Social capital is believed to play an important role in the functioning of community life across a variety of domains, ranging from the prevention of juvenile delinquency and crime, the promotion of successful youth development, and the enhancement of schooling and education to the encouragement of political participation. More recently, researchers have begun to apply the concept to explain variations in health status across geographic localities. In preliminary analyses, the higher the stocks of social capital (as indicated by measures of trust and reciprocity in social surveys), the higher appear to be the health achievement of a given area. Strengthening the social capital within communities may provide an important avenue for reducing socioeconomic disparities in health. PMID- 10681894 TI - Socioeconomic status and chronic stress. Does stress account for SES effects on health? AB - Socioeconomic status (SES) is an important predictor of a range of health and illness outcomes. Research seeking to identify the extent to which this often reported effect is due to protective benefits of higher SES or to toxic elements of lower social status has not yielded consistent or conclusive findings. A relatively novel hypothesis is that these effects are due to chronic stress that is associated with SES; lower SES is reliably associated with a number of important social and environmental conditions that contribute to chronic stress burden, including crowding, crime, noise pollution, discrimination, and other hazards or stressors. In other words, chronic stress may capture much of the variance in health and social outcomes associated with harmful aspects of lower social status. Low SES is generally associated with distress, prevalence of mental health problems, and with health-impairing behaviors that are also related to stress. Research targeting this hypothesis is needed to determine the extent to which stress is a pathway linking SES and health. PMID- 10681895 TI - Status, stress, and atherosclerosis: the role of environment and individual behavior. AB - Atherosclerosis induced by moderate hyperlipoproteinemia in group-housed cynomolgus monkeys differs significantly between animals of dominant and subordinate social status. The nature of this association also varies by sex, and in males, by stability of the social environment. Dominant males develop more extensive atherosclerosis than subordinates when housed in unstable, but not stable, social groups; in contrast, subordinate females develop greater atherosclerosis than dominants, and do so irrespective of the conditions of social housing. Experimental investigations reveal that the first of these associations (males) is mediated by concomitant sympathoadrenal activation and the second (females) by ovarian impairment associated with the stress of social subordination. We believe our findings offer clues to the neuroendocrine mediation of behavioral influences on coronary artery disease in humans. This is particularly true where these influences reflect asymmetries in the power or status relationships among individuals within similar social environments, or when dimensions of temperament or disposition give rise to such relationships. We propose that these data also may be informative regarding the pathophysiological sequelae of social stratification (in which disease incidence varies by class membership within populations), but only where social environments engendered by class inequalities exacerbate status-dependent behavioral differences among individuals within communities of associates. PMID- 10681896 TI - Stress responses in low-status jobs and their relationship to health risks: musculoskeletal disorders. AB - Conditions typical of many low-status jobs are known to induce elevated stress. In keeping with this, blue-collar workers show elevated psychophysiological stress levels both during and after work compared with workers in more stimulating and flexible jobs. Health-related behaviors, such as cigarette smoking and drug abuse, that are known to contribute to the social gradient in health, can be seen as ways of coping with a stressful work situation in order to get short-term relief. Negative emotional states associated with low-status jobs, combined with a lack of economic resources, are also likely to reduce the individual's motivation to seek proper medical treatment and, thus, increase the risk that transient symptoms develop into chronic illness. With regard to musculoskeletal disorders, it is well documented that physically monotonous or repetitive work is associated with an increase in neck, shoulder, and low back pain problems. However, recent studies also report an association between psychosocial factors and muscle pain syndromes. Possible mechanisms explaining these findings involve the assumption that psychological stress may induce sustained activation of small, low-threshold motor units that may lead to degenerative processes, damage, and pain. Analysis of short periods of very low muscular electrical activity (EMG gaps) shows that female workers with a high frequency of EMG gaps seem to have less risk of developing myalgia problems than do workers with fewer gaps. Stress induced by psychosocial conditions at work, which is usually more lasting than that resulting from physical demands, may prevent the individual from shutting off their physiological activation and reduces the time for rest and recovery. In the modern work environment, with strong emphasis on a high work pace, competitiveness, and efficiency, it is possible that lack of relaxation is an even more important health problem than is the absolute level of contraction or the frequency of muscular activation. PMID- 10681897 TI - Race, socioeconomic status, and health. The added effects of racism and discrimination. AB - Higher disease rates for blacks (or African Americans) compared to whites are pervasive and persistent over time, with the racial gap in mortality widening in recent years for multiple causes of death. Other racial/ethnic minority populations also have elevated disease risk for some health conditions. This paper considers the complex ways in which race and socioeconomic status (SES) combine to affect health. SES accounts for much of the observed racial disparities in health. Nonetheless, racial differences often persist even at "equivalent" levels of SES. Racism is an added burden for nondominant populations. Individual and institutional discrimination, along with the stigma of inferiority, can adversely affect health by restricting socioeconomic opportunities and mobility. Racism can also directly affect health in multiple ways. Residence in poor neighborhoods, racial bias in medical care, the stress of experiences of discrimination and the acceptance of the societal stigma of inferiority can have deleterious consequences for health. PMID- 10681898 TI - Pathways by which SES and ethnicity influence cardiovascular disease risk factors. AB - Little is known about pathways by which socioeconomic status (SES) translates into individual differences in cardiovascular disease (CVD) risk factors. Because the socioeconomic structure is not the same for all ethnic subgroups, the pathways that lead to the development of CVD risk factors may vary by both SES and ethnicity. We used data from a large national survey to examine the independent associations of two indicators of SES (education and income) and ethnicity with six primary CVD risk factors. We then used data on smoking that reflected a temporal sequence to examine the extent to which SES and ethnicity influenced smoking at three different time points, from smoking onset, to a serious quit attempt, to successful quitting. These analyses provide an understanding of the relationships between SES, ethnicity, and CVD risk factors and suggest that if the timing, focus, and content of intervention programs take pathways into account they will result in more successful outcomes. PMID- 10681899 TI - Psychosocial resources and the SES-health relationship. AB - Psychosocial resources, which include optimism, coping style, a sense of mastery or personal control, and social support, influence the relationship between SES and health. To varying degrees, these resources appear to be differentially distributed by social class and related to health outcomes. Such resources may partially mediate the impact of SES on health. For example, environments that undermine personal control may have an impact on chronic arousal and the corresponding development of disease, such as CHD. Psychosocial resources may also moderate the impact of SES on health. For example, a large number of positive social relationships and a few conflictual ones may buffer individuals against the adverse effects of SES-related stress. These psychosocial resources are moderately intercorrelated, and so a research strategy that explores their coherence as a psychosocial profile that promotes resilience to stress is tenable and merits empirical examination. The erosion of these resources as one moves lower on the SES scale and specific factors that contribute to such erosion are discussed. PMID- 10681900 TI - Do negative emotions mediate the association between socioeconomic status and health? AB - In this chapter, we examine the possibility that negative emotions contribute to the relationship between socioeconomic status (SES) and health. A model of the associations among SES, emotion, and health is presented first. We then review the evidence for this model, showing associations of SES with depression, hopelessness, anxiety, and hostile affect and cognition, and of these negative emotions with disease. Notably, most of the data supporting the model provide only indirect evidence that negative emotions serve as a key contributor to the proposed associations. We, therefore, conclude with recommendations for longitudinal research, especially in children, that will more directly and comprehensively examine negative emotions as possible mediators of the SES and health relationship. PMID- 10681901 TI - Social status and susceptibility to respiratory infections. AB - Adults and children of lower socioeconomic status (SES) are at higher risk for a wide range of communicable infectious diseases, especially respiratory infections. Greater risk for infectious illness among people with lower SES is thought to be attributable to increased exposure to infectious agents and decreased host resistance to infection. We summarize three studies that examine the prospective association of several markers of social status (unemployment, perceived and observed social status) with host resistance to upper respiratory infections. Unemployment was associated with increased susceptibility to infection in adult humans. Lower social status in male monkeys was also associated with increased susceptibility, as was lower perceived social status in humans. The association of social status and susceptibility was accounted for primarily by increased risk in the lowest social status groups. However, further increases in social status were associated with further decreases in susceptibility in both monkeys and humans. PMID- 10681902 TI - Sleep as a mediator of the relationship between socioeconomic status and health: a hypothesis. AB - This article discusses the hypothesis that the adverse impact of low socioeconomic status (SES) on health may be partly mediated by decrements in sleep duration and quality. Low SES is frequently associated with a diminished opportunity to obtain sufficient sleep or with environmental conditions that compromise sleep quality. In a recent study, we examined carbohydrate metabolism, endocrine function, and sympatho-vagal balance in young, healthy adults studied after restricting sleep to four hours per night for six nights as compared to a fully rested condition obtained by extending the bed-time period to 12 hours per night for six nights. The state of sleep debt was associated with decreased glucose tolerance, elevated evening cortisol levels, and increased sympathetic activity. The alterations in glucose tolerance and hypothalamo-pituitary-adrenal function were qualitatively and quantitatively similar to those observed in normal aging. These results indicate that sleep loss can increase the "allostatic load" and facilitate the development of chronic conditions, such as obesity, diabetes, and hypertension, which have an increased prevalence in low SES groups. PMID- 10681903 TI - Cardiovascular pathways: socioeconomic status and stress effects on hypertension and cardiovascular function. AB - In westernized societies there is a consistent and continuous gradient between the prevalence of cardiovascular disease (including both coronary heart disease and stroke) with SES, such that people from lower SES have more disease. Several studies have examined the roles of the major cardiovascular risk factors for explaining this gradient. There is a strong SES gradient for smoking, which parallels the gradient in disease, but the gradients for hypertension and cholesterol are weak or absent. Central obesity and physical inactivity may also be contributory factors. In the United States there is a strong association between SES and race, and it is suggested that the higher prevalence of hypertension and cardiovascular disease in blacks may be attributed to psychosocial factors, including those related to SES. The possible pathways by which SES affects cardiovascular disease include effects of chronic stress mediated by the brain, differences in lifestyles and behavior patterns, and access to health care. At the present time, the second of these is the strongest candidate; the effects of stress have been little studied. PMID- 10681904 TI - Public policy frameworks for improving population health. AB - Four conceptual frameworks provide bases for constructing comprehensive public policy strategies for improving population health within wealthy (OECD) nations. (1) Determinants of population health. There are five broad categories: genes and biology, medical care, health behaviors, the ecology of all living things, and social/societal characteristics. (2) Complex systems: Linear effects models and multiple independent effects models fail to yield results that explain satisfactorily the dynamics of population health production. A different method (complex systems modeling) is needed to select the most effective interventions to improve population health. (3) An intervention framework for population health improvement. A two-by-five grid seems useful. Most intervention strategies are either ameliorative or fundamentally corrective. The other dimension of the grid captures five general categories of interventions: child development, community development, adult self-actualization, socioeconomic well-being, and modulated hierarchical structuring. (4) Public policy development process: the process has two phases. The initial phase, in which public consensus builds and an authorizing environment evolves, progresses from values and culture to identification of the problem, knowledge development from research and experience, the unfolding of public awareness, and the setting of a national agenda. The later phase, taking policy action, begins with political engagement and progresses to interest group activation, public policy deliberation and adoption, and ultimately regulation and revision. These frameworks will be applied to help understand the 39 recommendations of the Independent Inquiry into Inequalities in Health, the Sir Donald Acheson Report from the United Kingdom, which is the most ambitious attempt to date to develop a comprehensive plan to improve population health. PMID- 10681905 TI - Socioeconomic status and health. Policy implications in research, public health, and medical care. AB - The role of public policy in research, public health, and medical care is discussed as well as the extent to which public policy has been informed by increased knowledge about the relationship between SES and health and to what extent policy has affected SES-related health disparities. Observations on current healthcare policy and recommendations for the future are offered. PMID- 10681906 TI - Solving the puzzle of socioeconomic status and health: the need for integrated, multilevel, interdisciplinary research. PMID- 10681907 TI - Social class differences in morbidity using the new U.K. National Statistics Socio-Economic Classification. Do class differences in employment relations explain class differences in health? PMID- 10681908 TI - Orthostatic blood pressure responses as a function of ethnicity and socioeconomic status: the ARIC study. PMID- 10681909 TI - Social class as a moderator of income effects on stress and health outcomes across nine years. PMID- 10681910 TI - Socioeconomic disparities in adult oral health in the United States. PMID- 10681911 TI - Income inequality, social trust, and self-reported health status in high-income countries. PMID- 10681912 TI - The Human Development Index and Per Capita Gross National Product as predictors of dental caries prevalence in industrialized and industrializing countries. PMID- 10681913 TI - Pathways between area-level income inequality and increased mortality in U.S. men. PMID- 10681914 TI - Education, income, wealth, and health among whites and African Americans. PMID- 10681915 TI - Income inequality and mortality in Canada and the United States. An analysis of provinces/states. PMID- 10681916 TI - For richer, for poorer, in sickness and in health: socioeconomic status and health among married couples. PMID- 10681917 TI - Determinants of health: testing of a conceptual model. PMID- 10681918 TI - The direct and indirect effects of metropolitan area inequality on mortality. A hierarchical analysis. PMID- 10681919 TI - Unbundling education: a critical discussion of what education confers and how it lowers risk for disease and death. PMID- 10681920 TI - Socioeconomic disparities in adolescent health: contributing factors. PMID- 10681921 TI - Effects of socioeconomic status and psychosocial stress on the development of the fetus. PMID- 10681922 TI - Social position, age, and memory performance in the Whitehall II Study. PMID- 10681923 TI - Social dominance and cardiovascular reactivity in preschoolers. Associations with SES and health. PMID- 10681925 TI - Racial differences in education, obesity, and health in later life. PMID- 10681924 TI - Markers of transition to adulthood, socioeconomic status of origin, and trajectories of health. PMID- 10681926 TI - Limitations to the use of education as an SES indicator in studies of the elderly. Confounding by cognition. PMID- 10681927 TI - Parenting behavior and emotional health as mediators of family poverty effects upon young low-birthweight children's cognitive ability. PMID- 10681928 TI - Socioeconomic factors and emergency pediatric ICU admissions. PMID- 10681929 TI - Socioeconomic status and health among elderly people in Sweden. PMID- 10681930 TI - A multivariate model of functional decline in the elderly: the differential influence of income versus education. PMID- 10681931 TI - Socioeconomic status, social support, age, and health. PMID- 10681932 TI - Measurement of social capital. PMID- 10681933 TI - Racism and health: segregation and causes of death amenable to medical intervention in major U.S. cities. PMID- 10681934 TI - Identifying social pathways for health inequalities. The role of housing. PMID- 10681935 TI - Transitions into poverty following job loss and the depression-reemployment relationship. PMID- 10681936 TI - Longitudinal effects of occupational, psychological, and social background characteristics on health of older workers. PMID- 10681937 TI - Social class and social cohesion: a content validity analysis using a nonrecursive structural equation model. PMID- 10681938 TI - The effect of job strain on ambulatory blood pressure in men: does it vary by socioeconomic status? PMID- 10681939 TI - Socioeconomic differences in social information processing and cardiovascular reactivity. PMID- 10681940 TI - Social environmental stress in indigenous populations: potential biopsychosocial mechanisms. PMID- 10681941 TI - Social status, anabolic activity, and fat distribution. PMID- 10681942 TI - Socioeconomic status as a correlate of sleep in African-American and Caucasian women. PMID- 10681943 TI - The differential effects of sleep quality and quantity on the relationship between SES and health. PMID- 10681944 TI - Stress responsivity and body fatness: links between socioeconomic status and cardiovascular risk factors in youth. PMID- 10681945 TI - Social class differences in maternal stress appraisal during pregnancy. PMID- 10681946 TI - The effects of race, gender, and education on the structure of self-rated health among community-dwelling older adults. PMID- 10681947 TI - Socioeconomic differences in measures of hostility. PMID- 10681948 TI - SES and oral health status in an elderly population. PMID- 10681949 TI - Hostility, coronary heart disease, and ischemia. The role of socioeconomic status. PMID- 10681950 TI - Education, infant health, and cigarette smoking. PMID- 10681951 TI - Morbidity and health--National Health Interview Survey, 1987. In generalized additive models. PMID- 10681952 TI - Neighborhood socioeconomic context and adult health. The mediating role of individual health behaviors and psychosocial factors. PMID- 10681953 TI - Goal-striving stress, social economic status, and the mental health of black Americans. PMID- 10681954 TI - Moderating and mediating effects of socioeconomic status, perceived peer condom use, and condom negotiation on sexual risk behavior among African-American and white adolescent females. PMID- 10681955 TI - Dissociative disruptions--psychological (psychosocial) pathways uncovered in the healing process. PMID- 10681956 TI - Does cognitive functioning mediate the well-documented link between education and functional disability in middle-aged adults? PMID- 10681957 TI - Socioeconomic status and health: a new explanation. PMID- 10681958 TI - A framework for evidenced-based reviews of interventions for supportive social environments. PMID- 10681959 TI - Social ordering in developing countries: does hierarchy have the same effect as in post-industrial nations? A look at Nepal. PMID- 10681960 TI - SES, Medicare coverage, and flu shot utilization among vulnerable women in the women's health and aging study. PMID- 10681961 TI - The association of race/socioeconomic status and use of Medicare services. A little-known failure in access to care. PMID- 10681962 TI - Natural and bioterrorist/biocriminal threats to food and agriculture. PMID- 10681963 TI - Agriculture and food security. PMID- 10681964 TI - The Soviet Union's anti-agricultural biological weapons. PMID- 10681965 TI - The threat posed by the global emergence of livestock, food-borne, and zoonotic pathogens. PMID- 10681966 TI - Contemporary global movement of emerging plant diseases. AB - Plant diseases are a significant constraint to agricultural productivity. Exotic plant diseases pose a continued threat to profitable agriculture in the United States. The extent of this threat has increased dramatically in the 1980s and 1990s due to the expansion of international trade in agricultural products and frequent movement of massive volume of people and goods across national boundaries. Introduction of new diseases has not only caused farm losses, but has also diminished export revenue since phytosanitary issues are linked to international commerce. Plant pathogens and their vectors have also moved across national boundaries, sometimes naturally and at other times influenced by the recent changes in trade practices. Sorghum ergot, Karnal bunt of wheat, potato late blight, and citrus tristeza are some of the most recent examples of enhanced importance of diseases due to the introduction of plant pathogens or vectors. PMID- 10681967 TI - Biological warfare training. Infectious disease outbreak differentiation criteria. AB - The threat of biological terrorism and warfare may increase as the availability of weaponizable agents increase, the relative production costs of these agents decrease, and, most importantly, there exist terrorist groups willing to use them. Therefore, an important consideration during the current emphasis of heightened surveillance for emerging infectious diseases is the capability to differentiate between natural and intentional outbreaks. Certain attributes of a disease outbreak, while perhaps not pathognomic for a biological attack when considered singly, may in combination with other attributes provide convincing evidence for intentional causation. These potentially differentiating criteria include proportion of combatants at risk, temporal patterns of illness onset, number of cases, clinical presentation, strain/variant, economic impact, geographic location, morbidity/mortality, antimicrobial resistance patterns, seasonal distribution, zoonotic potential, residual infectivity/toxicity, prevention/therapeutic potential, route of exposure, weather/climate conditions, incubation period, and concurrence with belligerent activities of potential adversaries. PMID- 10681968 TI - The U.S. Department of Agriculture Food Safety and Inspection Service's activities in assuring biosecurity and public health protection. PMID- 10681969 TI - Safeguarding production agriculture and natural ecosystems against biological terrorism. A U.S. Department of Agriculture emergency response framework. AB - Foreign pest introductions and outbreaks represent threats to agricultural productivity and ecosystems, and, thus, to the health and national security of the United States. It is advisable to identify relevant techniques and bring all appropriate strategies to bear on the problem of controlling accidentally and intentionally introduced pest outbreaks. Recent political shifts indicate that the U.S. may be at increased risk for biological terrorism. The existing emergency-response strategies of the Animal and Plant Health Inspection Services (APHIS) will evolve to expand activities in coordination with other emergency management agencies. APHIS will evolve its information superstructure to include extensive application of simulation models for forecasting, meteorological databases and analysis, systems analysis, geographic information systems, satellite image analysis, remote sensing, and the training of specialized cadres within the emergency-response framework capable of managing the necessary information processing and analysis. Finally, the threat of key pests ranked according to perceived risk will be assessed with mathematical models and "what if" scenarios analyzed to determine impact and mitigation practices. An infrastructure will be maintained that periodically surveys ports and inland regions for the presence of exotic pest threats and will identify trend abnormalities. This survey and monitoring effort will include cooperation from industry groups, federal and state organizations, and academic institutions. PMID- 10681970 TI - Guarding against natural threats and terrorist attacks. An industry perspective. PMID- 10681971 TI - The role of national animal health emergency planning. AB - Bioterrorism is one of many contingencies that we must be better prepared to prevent from adversely impacting our food security system. Education and awareness, prevention-based training, increased monitoring and surveillance, and greater focus on coordination between government agencies at both the federal and state levels will be necessary to develop a more prevention-based national food security system. The formation of the Steering Committee on National Animal Health Emergency Management is the beginning of a profound new process. The animal industry is driving the formation of a new national animal health emergency management system. It is designed to coordinate the emergency prevention and response activities of all partners, including government (state and federal), livestock producers, and veterinary practitioners. The goal is to develop a national coordinated strategy that places primary emphasis on preventing a broader spectrum of hazards from occurring throughout the food production chain. Coordination and integration of efforts are necessary with many different stakeholders, including government agencies other than U.S. Department of Agriculture. Through a proposed National Animal Health Emergency Management Council, all stakeholders can assist in the development of a national strategic plan that can better protect our food security system. PMID- 10681972 TI - Industry concerns and partnerships to address emerging issues. PMID- 10681973 TI - International economic considerations concerning agricultural diseases and human health costs of zoonotic diseases. PMID- 10681974 TI - The cost of disease eradication. Smallpox and bovine tuberculosis. AB - Although eradication is the ideal approach to reduce the economic and human health costs of disease, there may be both short- and long-term consequences. A $300 million effort succeeded in completely eradicating smallpox in less than ten years. The campaign was effective because variola virus produced acute illness, had no carrier stage or non-human reservoirs, and had an effective vaccine that was used in combination with international surveillance and public education. Bovine tuberculosis was completely eradicated in many U.S. herds at a cost of $450 million over 50 years using a "test and slaughter" program combined with meat inspection. Mycobacterium bovis often does not produce acute disease, persists in the carrier stage, has multiple non-human reservoirs, and easily crosses species. No effective vaccine or centralized global surveillance or eradication programs currently exist. Control measures result in significant economic losses. Smallpox eradication had limited economic consequences but has left much of world's population highly susceptible to zoonotic orthopoxviruses and to the use of smallpox as a biologic weapon. The primary threat of M. bovis exists in wildlife that share watering holes or pasture land with domestic stock. In the developed world, surveillance can minimize risks, but one-third of the world's population lacks effective agricultural and food safety programs, leaving them at substantial risk for zoonotic infection by M. bovis. PMID- 10681975 TI - Economic considerations of agricultural diseases. PMID- 10681976 TI - Regionalization's potential in mitigating trade losses related to livestock disease entry. PMID- 10681977 TI - Foreign animal disease agents as weapons in biological warfare. PMID- 10681978 TI - Tools and methods for protection of targets and infrastructures associated with food and agriculture industries. PMID- 10681979 TI - Infecting soft targets. Biological weapons and Fabian forms of indirect grand strategy. PMID- 10681980 TI - The first step toward building tools and methods for protection. PMID- 10681981 TI - Targeted immune design using RNA immunization. PMID- 10681982 TI - Sensitive and rapid identification of biological threat agents. PMID- 10681983 TI - United States of America v. Ray Wallace Mettetal, Jr. Preliminary observations. PMID- 10681984 TI - Terrorism overview. PMID- 10681985 TI - The changing biological warfare threat. Anti-crop and anti-animal agents. PMID- 10681986 TI - Trends in American agriculture. Their implications for biological warfare against crop and animal resources. AB - Current trends in American agriculture have changed the vulnerability to use of biological weapons against plant and animal resources. The major effect has been a requirement to look again at the model of the U.S. BW program of widespread dissemination of agent and look to attack models requiring much lower levels of resources. The U.S. biological warfare program models must take the effects of these major trends into account when considering the possible widespread dissemination of a biological agent. The models must also acknowledge the lowered levels or resources required to make such attacks given the modern trends in American agriculture. PMID- 10681987 TI - Agroterrorism. Agricultural infrastructure vulnerability. AB - The intentional contamination of animal feed to reduce the availability of animal derived human food or to infect human populations is seldom mentioned, but animal feed could be an easy target for bioterrorists. The period of delay between the contamination of the animal feed and adulteration of the human food product provides an additional degree of uncertainty about the source of the contamination and minimizes the possibility of apprehending the terrorist. The less obvious and more natural the source of biological contamination, the greater the likelihood that the animal feed contamination will be mistaken as a natural phenomenon. However, the problems related to managing natural food contamination and intentional food contamination remain the same. Rapid testing and separation of contaminated feed are important steps, followed by the more specific identification of the contaminant to determine the source of adulteration and/or the possibility of decontamination. At this time identification of the bioagents is dependent on the availability of antibody-specific test systems. The rapid development of specific antibodies for the development of sensitive and specific test kits is the key to identifying contamination and dealing effectively with the disposal or decontamination of the animal feed and, ultimately, preventing the contamination of animal-derived human food products. PMID- 10681988 TI - The need for a coordinated response to food terrorism. The Wisconsin experience. PMID- 10681989 TI - The threat of bioterrorism to U.S. Agriculture. PMID- 10681990 TI - Where have all my pumpkins gone? The vulnerability of insect pollinators. PMID- 10681991 TI - The role of pesticides in agricultural crop protection. PMID- 10681992 TI - The status and role of vaccines in the U.S. food animal industry. Implications for biological terrorism. AB - This paper was intended to highlight some of the disease agents that could be used effectively in acts of terrorism. In terms of vaccine countermeasures, we face situations on both ends of the spectrum--(1) we and other nations have not invested enough and have not been successful in developing or licensing any protective vaccines and (2) where vaccines are available but not commercially used due to current FAD policies we have not stockpiled them in sufficient doses should regular practices fail to contain an outbreak. It is hoped that this paper provokes additional thought and planning for those government agencies involved in the business of national food animal agricultural welfare. Vaccine technologies are available or are being developed to provide new and improved vaccines against these highly contagious agents. PMID- 10681993 TI - Exotic diseases of citrus. Threats for introduction? PMID- 10681994 TI - What should the G8 do about the biological warfare threat to international food safety? PMID- 10681995 TI - A domestic legislative agenda for improving food safety and safeguards from terrorist attacks on the U.S. food supplies and U.S. Agricultural interests. PMID- 10681996 TI - [From genetic screening to surgical cancer prophylaxis]. PMID- 10681997 TI - [Predictive genetic investigations. Individualization of diagnosis and treatment in families with multiple endocrine neoplasia type II]. AB - BACKGROUND AND OBJECTIVE: When multiple endocrine neoplasia type 2 (MEN2) is suspected, genetic tests are at the centre of screening procedures. It was the aim of this study to compare the diagnostic value of molecular biological investigations with that of conventional biochemical tests. PATIENTS AND METHODS: The study cohort consisted of all 144 patients cared for in our department since 1990 with the suspected diagnosis of MEN2 (evidence of a medullary thyroid carcinoma [MTC]), coexistence of two MEN2 tumours or a family history of MEN2. 14 of the 144 patients (from 12 families) were already known to have an hereditary MTC, while the remaining 130 had been referred for further diagnostic investigations. RESULTS: An hereditary MTC was diagnosed in 22 of the 130 patients, a sporadic MTC in 32, while no definitive classification was possible in 20 MTC patients without a positive family history and on whom no mutation analysis had been performed. MEN2 was excluded in 56 family members. All 22 patients with newly diagnosed MTC had abnormally high calcitonin levels. A germ line mutation in the RET proto-oncogene was found in 8 of the 9 families who had undergone molecular biological tests. The investigate results led to a thyroidectomy in 19 of the 22 patients with hereditary MTC; in all of them the surgical specimen showed C-cell hyperplasia and/o MTC. CONCLUSION: These results emphasize the importance of genetic tests in family screening. Preoperative measurement of calcitonin remains essential in MEN2 families in whom a germ-line mutation is not known. The choice of the appropriate diagnostic test must be individualized to the particular patients so that optimal results are obtained. PMID- 10681998 TI - [Decubitus ulcer in the terminal phase: epidemiologic, medicolegal and ethical aspects]. AB - BACKGROUND AND OBJECTIVES: Pressure sores usually result from insufficient preventive measures. They are particularly omnipresent among dying persons in geriatric care. This study deals with prevalence, risk factors and the significance of the nursing environment. PATIENTS AND METHODS: The prevalence of pressure sores among the dead was analysed in a prospective cross-sectional study based on 10,222 postmortem examinations in a crematorium in Hamburg. RESULTS: The overall prevalence of pressure sores from grades I to IV was 11.2% (grade I: 6.1%, grade II: 3%, grade III: 1.1%, grade IV: 0.9%). A final logistic regression model showed that pressure sores of Grade III or IV were associated with female gender, date of death in the summer, marasmus, stroke history, neurological disease in general, kidney disease, preceding traumatic events and nursery home residence at the time of death. More than half of all the grade IV cases were diagnosed among nursing home residents whereas those who had died in hospitals contributed to only 11.5% of all the grade IV cases (dead from private homes 34.4%). Nursing home residence was associated with female gender, marasmus and stroke history which predisposed to a higher rate of pressure sores. CONCLUSIONS: Nursing homes are confronted with the highest proportion of pressure sores among dying people when compared to hospitals or private home care. Failure to meet the standards of preventive action against pressure sores point to the shortfalls in the present public health sector and nursing home regulations as well as the medical responsibility for supervision of nursing care. Apart from established standards of care, medicolegal assessment of high-grade pressure sores should also take ethical considerations into account when considering maximum therapy goals among dying persons. PMID- 10682000 TI - [Alveolar echinococcosis: diagnosis]. PMID- 10681999 TI - [A patient with an ACTH-producing pituitary tumor with liver metastasis]. AB - HISTORY AND FINDINGS: A 57-year-old woman had an ACTH-producing pituitary adenoma twice resected, followed by bilateral adrenalectomy for recurrent hypercortisolism. She subsequently developed a secondary postadrenalectomy syndrome (Nelson's tumour) which required further surgery and radiotherapy. The patient now presented for elucidation of a space-occupying lesion in the liver, found incidentally on abdominal ultrasonography. INVESTIGATIONS: Immunocytochemistry of the liver biopsy revealed ACTH-producing cells that were structurally identical to the cells found in the specimen resected at the previous operation. Changes were also found in the lower thoracic vertebrae, suspicious of metastases, thus suggesting a metastasizing hypophyseal carcinoma. RESULTS AND COURSE: Resection of the primary tumour and subsequent radiotherapy had arrested the corticotropic, thyrotropic, and gonadotropic functions of the pituitary, which had been adequately treated by administration of the corresponding hormones. Ocreotide, bromocriptin or cytostatics were not given because of their reported doubtful efficacy. At the time of diagnosis of the malignancy a curative operation on the liver or palliative embolization of the liver metastases were not possible because of their number and size. The bone metastases were managed palliatively by radiotherapy. CONCLUSION: No curative treatment has been found for the 66 cases of hypophyseal carcinoma reported so far. Screening investigations in patients with operated pituitary adenoma with the aim of eliciting an early diagnosis of possible malignancy cannot, therefore, be recommended, particularly since renewed tumour growth and local invasiveness do not constitute criteria for the diagnosis of pituitary carcinoma. PMID- 10682001 TI - [Drug interactions with the cytochrome P-450 system]. PMID- 10682002 TI - [Medication prescriptions and costs in diabetic polyneuropathy]. PMID- 10682003 TI - [Lack of joy of publication in German biomedical research-- habilitation as a brake?]. PMID- 10682004 TI - Relief of acute pain: a basic human right? PMID- 10682005 TI - Allies or enemies? Evidence-based medicine and consumer choice. PMID- 10682006 TI - Hepatitis A, liver transplants and indigenous communities. PMID- 10682007 TI - Towards unity for health. PMID- 10682008 TI - Patient attitudes to commonly promoted medical interventions. AB - OBJECTIVE: To survey attitudes about three "best practice" medical interventions (hormone replacement therapy [HRT], thrombolysis for acute myocardial infarction [THROM] and coronary artery by-pass surgery [CABS]) in a sample of patients, and identify factors associated with those attitudes. SETTINGS: Metropolitan tertiary care hospital outpatient clinics (survey 1, April 1997), two general practice surgeries (survey 2, May 1997), and one general practice surgery (survey 3, October 1997). DESIGN: Patients completed a questionnaire while waiting for their clinical consultation. Attitude scores were measured on an 11-category Likert scale ranging from -5 (definitely would not) to +5 (definitely would) for acceptance of proposed medication or surgery. PARTICIPANTS: 85 (participation rate, 85%), 77 (94%) and 95 (97%) in surveys 1, 2 and 3, respectively. Surveys 1 and 2 constituted the primary study group (n = 162). Patients aged > or = 50 years or reporting heart disease were excluded from the HRT analyses; patients aged > or = 65 years were excluded from the THROM and CABS analyses. RESULTS: The median attitude scores for HRT (n = 58), THROM and CABS (n = 111) were -2.95 (95% CI, -5 to -2.1), -0.5 (95% CI, -0.9 to 0) and -0.1 (95% CI, -0.5 to +1.3), respectively. Decreasing the risk-benefit ratio fourfold for HRT in survey 3 (n = 68) increased the median score to -0.75 (95% CI, -2.3 to 0). CONCLUSIONS: Patients do not view favourably the risk-benefit ratio of the three surveyed medical interventions. These attitudes may present a major impediment to most primary prevention programs. PMID- 10682009 TI - Survival of patients with colorectal cancer detected by a community screening program. AB - OBJECTIVE: To determine survival rates for people with colorectal cancer detected through Bowelscan, a community screening program. DESIGN: Survey of data from local medical practitioners, and comparison with data from State cancer registries. SUBJECTS AND SETTING: 249 people with colorectal cancer detected after faecal occult blood screening in north-eastern New South Wales, 1987-1996. Follow-up was in 1998-1999. MAIN OUTCOME MEASURES: Five-year survival rates and relative survival ratios. RESULTS: Five-year survival rates for the screen detected cancer patients were 90% for those with Dukes' stage A cancers, 75% for Dukes' B, 52% for Dukes' C and 0 for Dukes' D (although one person with Dukes' D cancer was living at four-year follow-up at the end of the study). Because of the higher percentage of Dukes' A cases in the population whose cancer was detected through screening, the resulting five-year relative survival ratio was significantly better than for those recorded by New South Wales, South Australian and Queensland cancer registries: 0.82 (95% confidence interval, 0.74-0.90) compared to 0.59 (P < or = 0.001). CONCLUSIONS: The study supports the findings of three overseas randomised trials that screening reduces mortality from colorectal cancer. We estimate that screening 200,000 people would detect about 250 colorectal cancers and prevent as many as 55 deaths. PMID- 10682010 TI - Concordance between use of proton pump inhibitors and prescribing guidelines. AB - OBJECTIVE: To determine (i) the relationship between prescriptions for proton pump inhibitors (PPIs) and upper gastrointestinal conditions, and (ii) compliance with Pharmaceutical Benefits Scheme (PBS) prescribing guidelines for PPIs. DESIGN: Drug utilisation evaluation. SETTING: 800-bed metropolitan teaching hospital. PARTICIPANTS: 253 patients dispensed PPIs from the hospital pharmacy over five consecutive weeks (11 January to 15 February 1999). MAIN OUTCOME MEASURES: Recorded gastrointestinal conditions; previous trial of H2-antagonist therapy; compliance with PBS criteria for prescribing PPIs. RESULTS: Seventy patients (27.7%) had no appropriate upper gastrointestinal tract investigations, and 62 patients (24%) did not receive an adequate trial of H2-antagonist therapy before the commencement of a PPI. The major indications for use of PPIs in investigated patients were gastro-oesophageal reflux in 99 (54%) and peptic ulcer disease in 30 (16.4%). In only 57 patients (22.5%) did PPI prescriptions comply with PBS prescribing guidelines. Clinical indications that failed to meet prescribing criteria included milder forms of gastro-oesophageal reflux, gastritis/duodenitis, and non-specific dyspepsia with normal endoscopy results. CONCLUSION: Drug utilisation data indicate widespread use of PPIs outside current prescribing guidelines. Many patients have not had relevant investigations and/or an adequate trial of H2-antagonist therapy. These findings explain the considerable hospital expenditure on PPIs. PMID- 10682011 TI - Fulminant hepatitis A in indigenous children in north Queensland. AB - Since 1993, three Indigenous children in north Queensland have died of fulminant hepatitis A. Even if the children had been able to undergo liver transplantation, prolonged immunosuppressant therapy and the likelihood of opportunistic infections would inevitably have jeopardised any chance of long-term survival. As hepatitis A has become a leading infectious cause of death in young Indigenous children in north Queensland, hepatitis A vaccine has recently been introduced into the vaccination schedule for these children. PMID- 10682012 TI - General practice computerisation: lessons from the United Kingdom. PMID- 10682013 TI - An integrated electronic health record and information system for Australia? PMID- 10682014 TI - Integrated electronic health records and patient privacy: possible benefits but real dangers. PMID- 10682015 TI - 2020 vision: looking to the future. PMID- 10682016 TI - More than minding the machine. PMID- 10682017 TI - "Eyes-open optimism": one view of the future for physicians. PMID- 10682018 TI - Fallopian tube carcinoma detected by ThinPrep cytology smear. PMID- 10682019 TI - Computed tomography screening for coronary disease. PMID- 10682020 TI - Computed tomography screening for coronary disease. PMID- 10682021 TI - Computed tomography screening for coronary disease. PMID- 10682022 TI - Stable angina pectoris: treatment and referral options. PMID- 10682024 TI - Vancomycin-resistant enterococci and use of avoparcin in animal feed: is there a link? PMID- 10682023 TI - Vancomycin-resistant enterococci: seek and ye shall find. PMID- 10682025 TI - Vibrio cholerae in Victoria. PMID- 10682026 TI - Androgen treatment in women. PMID- 10682027 TI - Women's role and satisfaction in the decision to have a caesarean section. PMID- 10682028 TI - How best to fix a broken hip. PMID- 10682029 TI - Short and curly. PMID- 10682030 TI - [Prevention of venous thromboembolism in polytraumatized patients. Epidemiology and importance]. AB - OBJECTIVE: Unfractionated heparin, low-molecular-weight heparin (LMWH), mechanical compression, and vena cava filters are part of a large panel of chemical or physical methods proposed to trauma patients as prophylaxis against venous thromboembolism. This prophylactic strategy is based on a 1961 autopsy survey showing a 16.6% rate of pulmonary embolism in this population. The objective of this study was to assess the incidence of deep venous thrombosis (DVT) and pulmonary embolism (PE) in multiple trauma patients. METHODS: A Medline and Current Contents search was conducted for prospective studies including trauma patients with ISS > 9 whose incidence of DVT and PE was evaluated by contrast venography and/or duplex scan and by ventilation/perfusion lung scan and/or pulmonary angiography and/or autopsy, respectively. RESULTS: Twelve studies met the selection criteria for DVT. Among the global population of 2,374 trauma patients (14% of all admissions) 47% suffered lower limb injury and 17% had severe head injury. Overall incidence of DVT was about 20%. It was about 38% in patients without prophylaxis (range 2 to 61.5%) and about 13% in patients with prophylaxis (range 0.8 to 37%). Similar variations were observed for proximal DVT. In comparative studies (unfractionated heparin versus LMWH or versus mechanical compression devices), the incidence of thromboembolic events varied from 3.2 to 44% in patients given unfractionated heparin, 0.8 to 31% in those given LMWH, and 3.1% to 12% with mechanical compression. Thirteen studies met the selection criteria for PE and included an overall population of 4,245 trauma patients where the diagnosis of PE was suspected only if the patient had clinical signs. The incidence of PE and fatal PE was about 1.4% and 0.3% respectively. Only one study systematically studied the presence of PE and showed an incidence of 18.7% for PE in a population of 32 patients. DISCUSSION: The current literature reports wide variability in methodology, characteristics of the study population and prophylaxis. These differences explain the wide variability from one study to another in the risk factors for venous thrombosis identified by univariate and multivariate analysis. CONCLUSION: Current methodological imperfections make it impossible to correctly assess the incidence of DVT and PE in the multiple trauma population. Studies with a rigorous methodology using a precise stratification of the trauma injuries are required to determine the real risk for DVT/PE in trauma patients and to assess the impact of early systematic prophylaxis. PMID- 10682031 TI - [Administration of tobramycin aerosols in patients with nosocomial pneumonia: a preliminary study]. AB - OBJECTIVE: The aim of this study was to assess renal and respiratory tolerance of aerosolized tobramycin in intubated and mechanically ventilated patients with nosocomial pneumonia. PATIENTS AND METHODS: This was a multicenter, randomized, double-blind, placebo controlled study. Thirty-eight mechanically ventilated patients with documented nosocomial pneumonia were included. Patients treated with intravenous betalactam and tobramycin were randomly allocated to receive aerosolized tobramycin (6 mg/kg/day, n = 21) or placebo (n = 17). The aerosol was administered via a pneumatic nebulizer once a day for 5 days. RESULTS: Respiratory tolerance was good in all but two patients. No acute renal failure occurred. By day 10, 7 patients in the tobramycin group (35%) had been extubated versus 3 in the placebo group (18.5%, p = 0.18). By day 28, 6 patients had died (2 in the tobramycin group and 4 in the placebo group, p = 0.23). CONCLUSION: Aerosolized tobramycin was well tolerated in ventilated patients with documented nosocomial pneumonia. PMID- 10682032 TI - [Sudden-onset coma revealing neurosarcoidosis]. AB - BACKGROUND: Central nervous system involvement is uncommon in sarcoidosis, particularly in cases discovered in circumstances described in our case. CASE REPORT: A 38-year-old man with a history of pulmonary sarcoidosis diagnosed in 1991 was treated by budesonide. The clinical course was generally favorable until 1995 when the patient complained of rapid onset headache which suddenly progressed to coma. The patient died 26 days later. DISCUSSION: The course of the neurological manifestations was remarkable in this case of neurosarcoidosis. The anatomic localization was also unusual with 3 distinct foci outside the central nervous system. PMID- 10682033 TI - [Manchineel dermatitis]. PMID- 10682034 TI - [Troponin Ic in acute myocarditis in children]. PMID- 10682035 TI - [Basedow's disease and interferon for hepatitis C. Recurrence as Basedow's ophthalmopathy after interferon reintroduction]. PMID- 10682036 TI - [Flu and antiviral agents....]. PMID- 10682037 TI - [Anti-ulcer drugs. Indications in adults. French Agency of the Safety of Health Products]. PMID- 10682038 TI - [Indications for anti-ulcer drugs in adults]. PMID- 10682039 TI - [Impact: from nutrition to drug]. PMID- 10682040 TI - [The French pharmacovigilance system: structure and missions]. AB - THE NETWORK: The French pharmacovigilance system is composed of a network of 31 regional pharmacovigilance centers located in convenient proximity to health care professionals. CAUSALITY ASSESSMENT: A causality assessment method is compulsory for all persons involved in pharmacovigilance in order to assess the causal relationship between an adverse effect and one or more drugs. PHARMACOEPIDEMIOLOGY: If necessary, an additional evaluation of the causal relationship is performed using pharmacoepidemiology methods. THE NATIONAL COMMISSION: A technical committee and a National Commission of Pharmacovigilance centralizes at the Agence Franciase de Securite Sanitaire (or AFSSAPS) and assesses all data in order to provide consensual advise to the relevant authorities on necessary measures, to prevent or reduce a drug-related adverse effect. PMID- 10682041 TI - [The European pharmacovigilance: regulatory aspects]. AB - ESTABLISHMENT: The European pharmacovigilance system has been operating since 1995 when the European Agency for the Evaluation of Medicinal Products as well as two new European registration procedures were established. STRUCTURE: This system is very similar to the French organization and is based on a decentralized collection and validation of safety data by member states and a centralized evaluation and decision making process at the European Agency for the Evaluation of Medicinal Products performed by the Committee for Proprietary Medicinal Products (CPMP) and its Pharmacovigilance Working Party. A EUROPEAN SYSTEM: In light of the experience gained, the European pharmacovigilance system moved to an interactive system which relies on a close cooperation between member states ensuring the common evaluation and management of safety concerns. PMID- 10682042 TI - [Incidence and prevalence of adverse drug reactions]. AB - FUNDAMENTAL DATA: Estimation of incidence and prevalence rates of adverse drug reactions are necessary to assess the consequences and impact of these effects in a population. HOSPITAL STUDIES: During the last 20 years, several studies conducted in one or more departments of a single hospital have found incidence of hospitalizations induced by adverse drug reactions between 2 and 8%. Three meta analyses confirm these reports with incidence varying from 4.4 to 5.8% in different countries and from 5.2 to 8.2% and from 2.37 to 3.6% in the United States and Australia respectively. Two recent studies conducted by the French pharmacovigilance regional centers on a representative sample of medical departments in public hospitals found a prevalence of adverse drug reactions between 4.2 and 22.1% (according to the type of department and hospital) and a hospitalization incidence due to adverse drug reaction between 2.37 and 4.01%. COMMUNITY STUDIES: In community medicine, few data are available. In general medicine, the incidence would be 1.99 adverse effects per general practitioner per day. The number of serious adverse drug reactions has been estimated at 0.01 per general practitioner per day. These numbers show the magnitude of adverse drug reactions in terms of morbidity and mortality. PMID- 10682043 TI - [Drug information and prescription guidance: role of regional centers of pharmacovigilance]. AB - MISSIONS: Drug information is one of the missions of the French Regional Pharmaco Vigilance Centers together with the evaluation of adverse drug reaction (ADR) reports, expertise, teaching and research. SERVICES: Physicians and other health professionals call their regional center (the address and phone numbers are on the first pages of the Vidal, the French national drug compendium) for any kind of information pertaining to adverse drug reactions or proper use of drugs such as drug use in pregnancy or lactation. RELIABILITY: This information activity is successful because of the competence and impartiality of the regional centers which are based in university hospitals under an agreement signed with the French Medicines Agency. PMID- 10682044 TI - [Antibiotic strategy in an infectious diseases unit]. PMID- 10682045 TI - [Prevention of venous thromboembolism in a non-surgical medical ward. Proposed indications for low molecular weight heparin]. AB - OBJECTIVE: The efficiency of venous thromboembolism prophylaxis with low molecular weight heparins (LMWH) has not been established in non surgical patients, so their official preventive use has been limited in France since 1995 to surgery. However, a survey conducted in 5 university hospitals in non surgical patients showed that 21-29% of patients still received a LMWH prescription. It seemed necessary to define the medical conditions for which the practical use of these heparins would be justified. We contacted external experts to obtain a consensus by using the Delphi method. METHODS: The Delphi method, created by the "Rand Corporation" in the USA and used in medicine since the nineteen seventies, is based on a light logistic, with questionnaires been sent by mail with a feed back report A total of 48 experts were chosen by local staff teams in the 5 hospitals. For the 3 rounds, from March to October 1998, questions were devised by a multicentred staff team. RESULTS: Among the 48 experts contacted, 32 completed the 3 questionnaires, 7 of them did for 2, and 43 did for at least one questionnaire. The experts first defined a list of 12 risk or high risk situations and 11 aggravating factors. For any high risk situation, prescription is justified. For other cases, 2 risk situations are required, or one risk situation with at least 2 aggravating factors, to justify a prescription. If no risk situation is present, prescription is, according to experts, usually not justified. CONCLUSION: The maximal agreement defines the situations in which one use of low molecular weight heparins is proposed to prevent deep venous thrombosis in non surgical inpatients, in most current hospital situations and for more than 24 hours of hospitalization. Clinical trials are needed, to validate their effectiveness and define the optimal dose in these indications. To date, epidemiological studies should be conducted to evaluate the experts proposals by estimating risk factors for deep venous thrombosis. PMID- 10682046 TI - [Value of Holter ECG in the diagnosis of sleep apnea syndrome in patients with massive obesity]. AB - OBJECTIVE: Assess the diagnostic contribution of cyclic nocturnal variations in heart rate in sleep apnea syndrome. PATIENTS AND METHODS: Holter recordings performed in a population of 30 patients with massive obesity defined as a body mass index greater than 40 kg/m2 and sleep apnea syndrome defined by an apnea index greater than 5 apneas per hour were analyzed retrospectively. The control group was composed of 15 patients with massive obesity but without sleep apnea syndrome. High variability in nocturnal heart rate was assessed using a visual criterion defined as repeated episodes of progressive reduction in heart rate followed by a sudden acceleration reaching a difference of 30 bpm between the highest and lowest heart rate and occurring at least 5 times during one consecutive hour of recording. RESULTS: Increased nocturnal variability in heart rate was evidenced in all the patients with sleep apnea syndrome (30/30) but was not observed in any of the control subjects (0/15). CONCLUSION: These results suggest that Holter recordings can be a useful tool for the diagnosis of sleep apnea syndrome. PMID- 10682047 TI - [Panhypopituitarism secondary to pituitary metastases]. AB - BACKGROUND: Hypophyseal metastatic localizations are uncommon and rarely the first expression of a primary cancer. We report an exceptional case revealed by panhypopituitarism and diabetes insipidis. CASE REPORT: Brain MRI visualized an intra- and suprasellar tumoral formation found to be a cribiform adenocarcinoma. The primary tumor could not be identified. Despite radiotherapy, surgery and chemotherapy combining carboplatin and etoposide, the tumor progressed with the development of cervical and mediastinal nodes. The patient died one year after onset of the clinical signs. DISCUSSION: Diagnosis of hypophyseal metastasis is mainly based on indirect evidence: rapid course, invasion of neighboring structures. Optimal management of these rare tumors remains to be determined. PMID- 10682048 TI - [Recurrent infectious and metastatic cellulitis cause by Escherichia coli]. AB - BACKGROUND: We report a case of recurrent and metastatic infectious cellulitis caused by Escherichi coli. CASE REPORT: A 79-year-old man with a history of alcoholic cirrhosis and a myelodyplasia syndrome was hospitalized for skin rash and inflammatory edema of the right leg associated with bullous and necrotic lesions. Culture of a bulla puncture fluid grew E. coli. A two-drug intravenous antibiotic regimen and surgical cleansing led to a favorable outcome in 3 weeks. One week after withdrawal of the antibiotics, the patient developed a recurrent erythematous and inflammatory lesion of the right flank. Blood culture grew E. coli. The intravenous antibiotics were reinitiated immediately and provided rapid regression of the skin signs. Search for a urinary or digestive tract neoplastic focus was negative. DISCUSSION: E. coli cellulitis is a very uncommon usually fatal condition. Clinicians should be aware of a possible association with alcoholic cirrhosis. In case of recurrence, it is important to search for a digestive, hepatobiliary or urinary tract focus. Broad spectrum empirical antibiotic therapy must be initiated rapidly. Surgery is required in case of necrotizing cellulitis whatever the infectious agent. PMID- 10682049 TI - [Ketoacidosis as an initial sign of diabetes decompensated by a flare up of Plasmodium falciparum infection]. PMID- 10682050 TI - [Fatal multiple organ failure in a homozygous sickle cell patient]. PMID- 10682051 TI - [Sural nerve palsy after stripping of the saphenous vein. 3 cases]. PMID- 10682052 TI - [Carpal tunnel syndrome: is corticosteroid infiltration sufficient to decompress the median nerve?]. PMID- 10682053 TI - [Therapeutic management of HIV-infected subjects. Report of the expert group under the direction of Prof. J.F. Delfraissy, 1999]. PMID- 10682054 TI - [Therapeutic management of HIU-infected persons. Interview with A, Sobel]. PMID- 10682055 TI - [Association of colon cancer and Hodgkin disease]. PMID- 10682056 TI - [Subcutaneous caffeine poisoning]. PMID- 10682057 TI - [Renal and hypertensive complications of extracorporeal lithotripsy]. AB - HISTOLOGICAL AND FUNCTIONAL CONSEQUENCES OF ESWL: Extracorporeal shock wave litotripsy is now used for the treatment of about 90% of stones. Because of the nonpunctual delivery of energy into the stone, a small volume of renal parenchyma is injured, giving rise to a fibrous scar which can be visualized by morphological techniques such as magnetic nuclear resonance. Isotopic techniques point out a 15% reduction of renal plasma flow on the side of the litotripsy. For a majority of patients, this alteration is transient. HYPERTENSION: In a few cases, abrupt onset of transient hypertension has been reported in clear relation with a compressive perirenal hematoma. The causal effect of ESWL on late occurrence of permanent hypertension is however still uncertain, probably because of the difficulty to show that this occurrence is not related to the older age of the patient alone. The FDA sponsored multicentric study begun in 1993 should solve this issue in the future. PATIENTS AT RISK: Recent articles suggest that altered renal function prior to ESWL would predict late occurrence of hypertension and worsening of renal failure. Furthermore, age and the resistance index of arcuate or interlobular renal arteries (measured by Doppler) could help to screen the patients at risk of developing hypertension. Practical attitude: In practice, renal function and blood pressure should be carefully monitored in patients aged over 60 and/or who have a serum creatinine > 300 mumol/l. PMID- 10682058 TI - [Venous thromboembolism in pregnancy]. AB - A DUAL CHALLENGE: Pregnancy is a physiological state favoring the development of venous thromboembolism and sometimes discloses a coagulation disorder. Due to the presence of the fetus, suspected venous thromboembolism in a pregnant woman raises a dual challenge for the clinician: confirmation of the clinically suspected diagnosis using imaging techniques exposing the fetus to as little radiation as possible, and adapted anticoagulant therapy taking into account the teratogenic risk. MILD TO MODERATE DISEASE: Excepting exceptionally severe cases, the only validated long-term treatment is continuous infusion heparin. However, because of the difficulties inherent in the use and control of this type of administration, most clinicians prefer low-molecular-weight heparins (LMWH) although these pharmaceutical products have not acquired official approval for this indication. PREVENTION: The optimal therapeutic approach for prevention of venous thromboembolism in a pregnant woman with an acquired or hereditary coagulation disorder or a history of venous thromboembolism remains to be defined. New clinical trials are needed to validate the use of LMWH in this indication and determine the therapeutic approach in certain risk situations and at delivery. PMID- 10682059 TI - [Multifactorial cardiovascular risk]. AB - IMPORTANCE OF RISK ASSESSMENT: The prevention of cardiovascular disease is a major public health goal. Cardiovascular diseases are the number one cause of mortality in industrialized countries and account for an important part of health care expenditures. In this context, assessment of the cardiovascular risk for a given subject based on epidemiological data and individual risk factors can be used to determine his/her risk of ischemic heart disease or stroke. THE FRAMINGHAM FORMULA: The most widely used assessment method is the Framingham formula which integrates age, sex, blood pressure, smoking habits and presence or not of diabetes. This formula gives an objective, reproducible estimation of the cardiovascular risk and is a useful tool for therapeutic rationale and primary and secondary prevention. INTEREST AND LIMITATIONS: This new global approach to the individual patient has interesting practical and economic implications but remains imperfect due to certain limitations (other risk factors not taken into account because they are difficult to quantify or occurred recently). For daily practice however, it provides a useful tool appreciated by clinicians and patients. PMID- 10682060 TI - [Effect of pregnancy, menopause and hormone substitution therapy on disseminated systemic lupus erythematosus]. AB - EXACERBATIONS DURING PREGNANCY: Clinical and experimental data have clearly evidenced the influence of hormones on the course of systemic lupus erythematosus. In prospective studies of pregnant women, an exacerbation is observed in 57% to 60% of the cases. It can be severe in 10% of the cases and occur in the post partum in 7%. For most patients, the exacerbation is moderate and has no unfavorable effect on the outcome of pregnancy. In case of renal involvement, it is difficult to differentiate an intricated HELPP syndrome. MARKERS AND RISK FACTORS: Low complement and elevated anti-DNA levels are distinctive markers. Earlier renal involvement and hypertension are important prognostic factors, particularly when the lupus begins during pregnancy. However, when serum creatinine is lower than 100 mumol/l at pregnancy onset in patients in remission, pregnancy does not alter renal function. An association with antiphospholipid antibodies increases the risk for the fetus and the kidney function. TREATMENT: Optimal treatment remains to be defined. Commonly, patients are given aspirin, heparin in case of a history of thromboembolism, or both. The rate of success currently exceeds 70%. The risk of thromboembolism in the peri or post partum period requires anticoagulant treatment. Outside pregnancy: Ovulation induction raises two risks: triggering a lupus flare-up and thrombosis, particularly for patients with antiphospholipid antibodies. The influence of menopause and hormone replacement therapy remains poorly understood. PMID- 10682061 TI - [Antibiotic strategies in an intensive care service]. PMID- 10682062 TI - Water channel AQP-1 in the primary cell culture of rat peritoneum. AB - To analyze the regulation of water channels in the peritoneum, we tried to establish a primary mesothelial cell culture system. Male Sprague-Dawley rats weighing about 250 g were anesthetized, and 10 mL of phosphate-buffered saline (PBS) containing 0.25% trypsin and 1 mmol/L ethylenediamine tetraacetic acid (EDTA) was infused into the peritoneal cavity for 15 minutes. Sediments from the recovered fluid were cultured in medium M199 supplemented with 10% fetal bovine serum (FBS). The culture was succeeded 4-6 times before experiments commenced. After exposure to the test medium, RNA was extracted and subjected to reverse transcriptase polymerase chain reaction (RT-PCR) for 10-19 cycles, then was measured by Southern blot analysis with a digoxin-labeled probe. Cultured cells were positively stained with mouse monoclonal anti-cytokeratin antibody, confirming their characteristics as mesothelial cells. Aquaporin-1 (AQP-1) message in the cultured cells increased with increases in glucose and mannitol concentrations when beta-actin message was used as an internal control. Tranexamic acid effected no change in AQP-1 message in the cultured mesothelial cells. This system offers potential as a simple approach to test the effects of osmolytes, cytokines, and vasoactive hormones on aquaporin expression and water transport in the peritoneum. PMID- 10682063 TI - Effects of chlorpromazine or diltiazem given intraperitoneally alone or in combination on peritoneal transport of solute and water. AB - Calcium channel blocker given intraperitoneally (i.p.) in rats was reported to increase urea D/P ratio without protein loss. Chlorpromazine (CP) given i.p. in humans was reported to increase ultrafiltration (UF) and urea clearance. We studied the effects of i.p. Diltiazem (DZ) (15 mg/kg) and i.p. chlorpromazine (0.25 mg/L dialysate)--given alone or in combination--on urea D/P ratio, dialysate protein (Dpro), glucose concentration (Dg), UF, and drainage volume (Vd). Six male Sprague-Dawley rats were studied. The rats underwent 21 consecutive 30-minute exchanges with 15 mL of 1.5% of Dianeal solution (Baxter Healthcare Inc., Deerfield, Illinois, U.S.A.). DZ or CP was added to the dialysis solution during exchanges 4-6 and 10-12. During exchange 16-18 both DZ and CP were added to the dialysis solution. Exchanges 1-3, 7-9, 13-15, and 19-21 were control exchanges performed with 1.5% Dianeal solution alone. The mean weight of the rats was 541.6 +/- 44 g. The animals' blood pressure remained stable during the study period. An increase in D/Purea ratio was observed with DZ, with CP, and with the two drugs in combination, without increase in dialysate protein loss. An increase in UF with a decrease in D/D0 was observed with DZ, with CP, and with the two drugs in combination, suggesting a mechanism other than osmotic gradient- such as increased blood flow or decreased surface tension. PMID- 10682064 TI - Decreased in vitro formation of AGEs with extraneal solution compared to dextrose containing peritoneal dialysis solutions. AB - Extraneal peritoneal dialysis (PD) solution (Baxter Healthcare, Deerfield, Illinois, U.S.A.) contains glucose polymer (icodextrin) as an osmotic agent in place of dextrose. We investigated the ability of Extraneal to form advanced glycation end products (AGEs) in vitro compared to standard PD solutions containing dextrose. Extraneal, Dianeal PD-2 [1.5%, 2.5%, or 4.25% dextrose (Baxter Healthcare)], or phosphate buffered saline (PBS) were incubated for 45 days with human serum albumin (HSA) or type IV collagen. AGE formation was measured by spectrofluorometry using excitation at 350 nm and emission at 430 nm. Solutions were also incubated with collagen affixed to plastic, simulating matrix collagen in the peritoneal membrane. In addition, AGE formation was assessed using icodextrin metabolites (maltose, maltotriose, and maltotetraose) at concentrations normally found in the plasma of patients treated using icodextrin. For PD solutions incubated with albumin, the relative order of AGE formation was: 4.25% dextrose > 2.5% dextrose > 1.5% dextrose > Extraneal. For incubations with collagen (in solution or affixed to plastic), AGE formation was greatest for 4.25% dextrose, intermediate for Extraneal and 2.5% dextrose, and lowest for 1.5% dextrose. Incubation of icodextrin metabolites with albumin for 45 days did not result in appreciable AGE formation. These results confirm that solutions containing icodextrin result in less in vitro AGE formation than do high dextrose solutions. The results also suggest that Extraneal may lead to improved solution biocompatibility in vivo. PMID- 10682065 TI - Effect of icodextrin peritoneal dialysis solution on cell proliferation in vitro. AB - Peritoneal dialysis solutions containing icodextrin are ideal for providing sustained ultrafiltration during long dwells, and they have replaced high glucose for long dwells in some patients. The biocompatibility of these solutions, especially in regard to glucose degradation products, has not been studied in depth. The object of this study was to compare the effects of commercially available dextrose-containing dialysis solutions to those of icodextrin containing solutions on fibroblast proliferation in vitro. We measured the effect of solutions on cell growth by exposing murine fibroblasts to pH-adjusted test solutions mixed with culture medium, and by comparing cell growth to growth in culture medium only. No statistical difference was observed in the growth of cells exposed to heat-sterilized Extraneal [7.5% icodextrin (Baxter Healthcare, Deerfield, Illinois, U.S.A.)], heat-sterilized Dianeal [1.5% dextrose (Baxter Healthcare)], or filter-sterilized Dianeal [4.25% dextrose (Baxter Healthcare]. Also, no difference was observed in the growth of fibroblasts exposed to heat sterilized Extraneal or to filter-sterilized Extraneal, but heat-sterilized Dianeal [4.25% dextrose (Baxter Healthcare)] caused a significant reduction in cell growth. Glucose degradation products (GDPs) are known to contribute to reduced cell growth in vitro. Extraneal had lower levels of the GDP acetaldehyde compared to Dianeal (2.5% or 4.25% dextrose). The results demonstrate enhanced in vitro biocompatibility characteristics for Extraneal, possibly related to low GDP levels in Extraneal. PMID- 10682066 TI - Peritoneal fluid kinetics: comparison between polyglucose solution and albumin solution. AB - Polyglucose (PG) solution has been shown to be capable of inducing peritoneal ultrafiltration despite its hypo-osmolality. However, the mechanism of osmosis by PG is not clear. In this study, we compared the fluid kinetics of albumin (ALB) solution (thought to be an ideal solution that should induce ultrafiltration through colloid osmosis) and PG solution. A 4-hour dwell study with frequent sampling was conducted in Sprague-Dawley rats (six rats in each group). The study used 25 mL of dialysate buffer, 10% ALB dialysis solution, 15% ALB dialysis solution, 20% ALB dialysis solution, or 7.5% PG solution, with 131I albumin as an intraperitoneal volume marker. All solutions were prepared by adding ALB or PG to a base dialysis solution (without osmotic agent). The initial osmolality values of the five solutions were 250 mOsm/kg, 284 mOsm/kg, 300 mOsm/kg, 320 mOsm/kg, and 280 mOsm/kg, and the dialysate drainage volumes at 4 hours were 17.0 +/- 0.8 mL, 22.4 +/- 0.8 mL, 25.4 +/- 0.6 mL, 27.3 +/- 0.9 mL, and 26.3 +/- 0.6 mL (buffer, 10% ALB, 15% ALB, 20% ALB, and 7.5% PG groups, respectively). The higher initial osmolality in the ALB groups was partially due to the sodium content in the ALB powder. The intraperitoneal volume was decreasing in the 10% ALB group, rather stable in the 15% ALB group, but slowly increasing in the 20% ALB group. In the PG group, intraperitoneal volume decreased initially and then started to increase after 2 hours. This pattern was closely related to the increase in the dialysate osmolality (to higher than plasma level). At 4 hours, the dialysate osmolality was significantly higher (and higher than plasma level) in the PG group as compared to all the ALB groups. No differences in peritoneal fluid absorption rate were observed among the four treatment groups. In the 15% ALB and 20% ALB groups, the transcapillary ultrafiltration rate (Qu) was lower in the later part of the dwell than in the initial part of the dwell; in the PG group, the opposite pattern was observed. Our results suggest that the osmosis of albumin dialysis solution is different from the osmosis of polyglucose solution. Polyglucose solution induces net ultrafiltration only when the dialysate osmolality increase to higher than plasma level, suggesting that degradation of polyglucose may be important for effective ultrafiltration. PMID- 10682067 TI - Changes in peritoneal membrane after continuous ambulatory peritoneal dialysis--a histopathological study. AB - Peritoneal membrane changes in continuous ambulatory peritoneal dialysis (CAPD) patients have been widely described but poorly classified. Our aim was to identify the morphological changes occurring after CAPD treatment. To this end, 17 biopsies of parietal peritoneum (1 cm in diameter) were withdrawn at least 5 cm from the catheter entry hole and stained with Van Gieson, hematoxylin-eosin, trichrome, and some immunohistochemical stains: keratin, vimentin, CD34, CD20, CD4, CD8, desmin, and collagen IV. The morphology of mesothelium, vessels, and basement membrane (BM) of mesothelium and vessels, the presence of inflammatory cells, fibrin, and calcifications, and the distribution and thickness of submesothelial tissue were evaluated. Patients were divided into three groups according to the thickness of the sclerotic band replacing mesothelium: group 1, band up to 40 microns; group 2, band less than 40 microns; group 3, no sclerotic band. The main histopathological alterations noted were: loss of mesothelium; sclerotic alteration of vessels or duplication of BM; presence of myofibroblasts; and presence of inflammatory cells (sparse, focal, or perivascular), mainly represented by macrophages and CD4+ lymphocytes. No significant qualitative differences were observed between the three groups. In conclusion, the variable histological changes in peritoneal membrane suggest a routine peritoneal biopsy in any surgical procedure to better understand pathological changes in the course of CAPD treatment. PMID- 10682068 TI - Evaluation of the effect of uremia on peritoneal permeability in an experimental model of continuous ambulatory peritoneal dialysis in anephric rats. AB - Anephric rats were maintained on continuous ambulatory peritoneal dialysis (CAPD). Peritoneal permeability was assessed during a standard 4-hour peritoneal equilibration test (PET) performed with Dianeal 3.86% (Baxter Healthcare, Deerfield, Illinois, U.S.A.). The effect of uremia on peritoneal permeability was evaluated in an experimental protocol in which each animal served as its own control. In each rat, PET1 (control) was performed before removal of kidneys and PET2 (uremia) was performed four days after removal of kidneys. Net ultrafiltration during a 4-hour exchange with Dianeal 3.86% was higher during PET1 (3.8 +/- 2.3 mL) than during PET2 (-1.3 +/- 3.3 mL), p < 0.05. Peritoneal permeability to urea and glucose was similar in both series. Transperitoneal equilibration of creatinine concentration was faster in uremic animals: D/P at 4 hours was 0.94 +/- 0.06 during PET2 versus 0.77 +/- 0.08 during PET1, p < 0.001. The opposite difference was seen for total protein: D/Px 1000 after a 4-hour dwell was 51.4 +/- 19.8 during PET2 versus 70.3 +/- 12.9 during PET1, p < 0.05. Our results show that uremia modifies the permeability of the peritoneum to both water and solutes. PMID- 10682069 TI - Peritoneal kinetics of cancer antigen 125 in peritoneal dialysis patients: the relationship with peritoneal outcome. AB - Cancer antigen 125 (CA125) is a mesothelial product that has been directly related with mesothelial bulk in peritoneal dialysis (PD) patients. Here, we evaluate CA125 levels in peritoneal effluent over time on PD, and relate them to changes in peritoneal function. We analyzed 27 peritoneal kinetic studies in 20 stable PD patients. Three patients dropped out of PD for peritoneal membrane failure after the last kinetic study, and six patients required a peritoneal rest period as treatment for membrane failure type I. We recorded the standardized daily ultrafiltration capacity, net ultrafiltration during the kinetic study, peritoneal mass transfer coefficients, time from onset of PD, and incidence of peritonitis prior to the study. A linear increase in CA125 levels over time was observed, and a strong correlation appears among the levels at different dwell times (r: 0.85-0.98, p < 0.05). At 180 minutes, the mean CA125 concentration was 48.5 +/- 39.7 U/mL. We observed significant differences in CA125 levels in effluent between the group of patients who later required a peritoneal rest period and the group of stable patients (27.7 +/- 26.3 U/mL vs 55.7 +/- 41.5 U/mL respectively, p < 0.05). Patients who left PD showed lower CA125 levels in effluent (31.4 +/- 30.6 U/mL vs 52.3 +/- 41.1 U/mL, p < 0.1). No correlation was seen between CA125 levels in effluent and time on PD, episodes of peritonitis, accumulated days of peritoneal inflammation, ultrafiltration capacity, or urea and creatinine mass transfer coefficients (MTCs). In conclusion, we believe that serial determinations of peritoneal effluent CA125 levels may help in the early identification of patients who show abnormal responses to peritoneal dialysis or its complications. PMID- 10682070 TI - Evaluation of changes in serum and dialysate levels of cancer antigen 125 in stable continuous ambulatory peritoneal dialysis patients. AB - To estimate the relationship between changes in the concentration of cancer antigen 125 (CA125) and peritoneal membrane kinetics, the permeability characteristics of 44 continuous ambulatory peritoneal dialysis (CAPD) patients who had been treated with peritoneal dialysis for at least six months were prospectively evaluated. Twenty-seven males (age 66 +/- 6 years, duration of CAPD 35.5 +/- 29 months) and seventeen females (age 63.7 +/- 9 years, duration of CAPD 47.7 +/- 32 months) were evaluated. Peritoneal equilibration test (PET) data and Adequest (Baxter Healthcare Corporation, Deerfield, Illinois, U.S.A.) data were analyzed in all patients over a 12-month period, while CA125 levels were measured in blood and dialysate samples. No statistically significant correlations were seen between the patients' age, sex, or peritonitis incidence rates, and serum and dialysate levels of CA125. Dialysate-to-plasma ratio (D/P) of small solutes at 0 and 240 minutes also showed no statistical correlation. Statistical analysis revealed a statistically significant negative correlation (r = -0.33, p = 0.035) between dialysate CA125 and duration of CAPD. The statistically significant difference found between dialysate CA125 concentrations at 0 minutes and 240 minutes (2.32 +/- 1.3 U/mL vs 9.08 +/- 6.8 U/mL, p < 0.0001), means that CA125 concentration increases with longer dwell time. These results suggest that the duration of CAPD clearly affects dialysate CA125 concentrations. CA125 may therefore be used as a useful marker to evaluate the mesothelial cell mass in longitudinal follow-up. PMID- 10682071 TI - Clinical importance of intraperitoneal pressure in peritoneal dialysis and measures to counteract its effect on net ultrafiltration. AB - Experiments in animals and in humans have shown that fluid loss from the peritoneal cavity to the body increases with large increments in the intraperitoneal hydrostatic pressure (IPP). We have demonstrated previously that much of this fluid loss occurs to the abdominal wall and is driven by the hydrostatic pressure gradient (i.p. pressure-skin pressure) that develops across the wall whenever therapeutic or pathologic volumes of fluid reside in the cavity. We hypothesized that eliminating the pressure difference across the wall by applying an equal and opposite pressure [abdominal counterpressure (ACP)] would decrease fluid movement into the wall and decrease fluid movement from the cavity. In addition, we hypothesized that net ultrafiltration or net fluid recovery would increase with ACP. To address these hypotheses, we dialyzed rats for 3 hours in the supine position at constant levels of IPP (4, 6, and 8 cmH2O) with isotonic or hypertonic dialysis solutions containing a protein marker of fluid movement. We measured total fluid loss, fluid marker concentration in the abdominal wall, and lymph flow. In separate animals, we repeated the experiments with ACP. Total fluid loss as determined by protein clearance and fluid marker deposition in the abdominal wall was decreased in all experiments. Lymph flow was unchanged by ACP. While ACP increased the net fluid recovery in isotonic dialysis, no change was observed in the hypertonic case. Analogous experiments were carried out in six dialysis patients with or without ACP during a 4-hour dialysis with 1.5% dextrose solution performed in the supine position at i.p. hydrostatic pressure of 4-6 cmH2O. No significant difference was noted in the measured net ultrafiltration between control and ACP studies. We conclude that the careful application of ACP does decrease fluid loss (particularly to the abdominal wall) during isotonic or hypertonic dialysis in the rat. However, ACP results in improved fluid recovery only with isotonic dialysis in rats and has no effect on the recovery of fluid during peritoneal dialysis in humans. PMID- 10682072 TI - Vasodilatation by intraperitoneal addition of nitroprusside is not a model for high peritoneal transport. AB - It has been suggested that the peritoneal capillary is the major determinant for peritoneal fluid transport and that an increase in the number of peritoneal capillaries may be the reason for an increased peritoneal transport rate. In this study, we investigated the impact on peritoneal fluid and solute transport of vasodilatation (which may increase the number of perfused capillaries) by intraperitoneal addition of nitroprusside. A 4-hour dwell was performed in four groups of Sprague-Dawley rats (4-8 rats in each group) with 131I albumin as an intraperitoneal volume marker. Nitroprusside at concentrations of 0% (control), 0.0005%, 0.001%, and 0.002% was added to the 3.86% glucose solution before intraperitoneal infusion. Net ultrafiltration (NUF) was 13.2 +/- 0.6 mL, 14.5 +/- 0.7 mL, 13.4 +/- 0.9 mL, and 6.3 +/- 2.0 mL for the control group and the 0.0005%, 0.001%, and 0.002% nitroprusside groups, respectively. No significant differences in peritoneal fluid absorption rate (KE) were observed among the four groups. The intraperitoneal volume, transcapillary ultrafiltration rate (Qu), and peritoneal small-solute transport rate (glucose and urea) as assessed by diffusive mass transfer coefficient were significantly higher or tended to be higher in the 0.0005% nitroprusside group as compared to control. However, with further increase in the dose of nitroprusside, these parameters started to decrease as compared to the 0.0005% nitroprusside group, and NUF was significantly lower in the 0.002% nitroprusside group, conceivably suggesting a reduction in blood flow owing to the systemic effects of nitroprusside. Our study showed that: (1) a low dose of nitroprusside increased the peritoneal small solute transport rate, but did not decrease peritoneal transcapillary ultrafiltration rate; (2) although a high dose of nitroprusside decreased peritoneal fluid removal, the decrease was not due to an increased small-solute transport rate, but more likely to a significant drop in peritoneal blood flow; (3) vasodilatation by nitroprusside did not change peritoneal fluid absorption rate. Overall, vasodilatation does not seem to produce a pattern of peritoneal fluid kinetics similar to that seen in high transporters, who have a higher peritoneal transport rate, lower transcapillary ultrafiltration rate, and higher peritoneal fluid absorption rate. PMID- 10682073 TI - Use of bolus intraperitoneal iron dextran in continuous ambulatory peritoneal dialysis or continuous cyclic peritoneal dialysis patients receiving recombinant human erythropoietin. AB - Impaired erythropoiesis in continuous ambulatory peritoneal dialysis (CAPD) or continuous cyclic peritoneal dialysis (CCPD) patients receiving recombinant human erythropoietin (rHuEPO) is most often secondary to iron deficiency, either as a result of poor intestinal absorption or failure to take oral supplements as prescribed. The inconvenience of giving intravenous (i.v.) iron dextran (ID) to CAPD/CCPD patients precluded its use in this population. We therefore examined the efficacy of bolus intraperitoneal (i.p.) iron dextran (1000 mg) on erythropoiesis in a pilot study of 14 CAPD/CCPD patients. The patients ranged in age from 23-81 years, and all had iron deficiency (transferrin saturation 6%-23%; mean: 15.2% +/- 1.34%). Of the 14 patients studied, 13 were receiving rHuEPO. Pre treatment hematocrit (Hct) ranged from 21%-38% (mean: 30.2% +/- 1.37%). After infusion of 2 L Dianeal (Baxter Healthcare Corp., Deerfield, Illinois, U.S.A.), 500 mg of undiluted ID was administered directly into the Tenckhoff catheter and subsequently flushed with 30 mm3 normal saline. The peritoneal dialysis (PD) exchange containing ID then dwelled for a period not < 6 hours before standard PD resumed. A second 500 mg dose ID was given to each patient by the same protocol 3 86 days later (mean: 14 days). No complications were seen. No patient complained of abdominal pain or other subjective symptoms during infusion or during the dwell. Repeat iron studies done 1-7 months post ID (mean: 2.8 months) showed a 1.1-fold to 4.9-fold increase (mean: 1.4-fold) in mean iron levels (40.4 +/- 3.9 mg/dL versus 57.5 +/- 5.5 mg/dL, p = 0.036); a 1.1-fold to 5.2-fold increase (mean: 1.6-fold) in mean transferrin saturation (15.2% +/- 1.3% versus 24.5% +/- 2.6%, p = 0.008); a 1.01-fold to 1.60-fold increase (mean: 1.12-fold) in mean Hct (30.2% +/- 1.37% versus 33.8% +/- 1.5%; p = 0.042). The mean dose of rHuEPO was statistically unchanged (170.0 +/- 47.4 U/kg body weight versus 178.8 +/- 49.6 U/kg body weight per week; p = 0.841). Peritoneal equilibration test (PET) score 1-4 months post ID (mean: 2 months) was 0.778 +/- 0.02 compared with a PET score at baseline of 0.767 +/- 0.03 (p = 0.734). No significant delta was observed in blood urea nitrogen (BUN) or creatinine values. We conclude that use of bolus i.p. ID is safe, effective, and convenient, and demonstrates no short-term negative effect on peritoneal membrane integrity. Long-term effects have yet to be determined. PMID- 10682074 TI - Correction of sodium sieving for diffusion from the circulation. AB - Transcellular water transport (TCWT) can be estimated by Na- sieving. However, the assumption that the initial Na+ dialysate concentration (D0) is equal to the initial plasma concentration (P0) is not true for each patient. The difference leads to Na+ diffusion from the circulation to the dialysate, which diminishes the Na+ sieving. A model was developed to distinguish transcellular water transport from Na+ diffusion. We previously found evidence that the mass transfer area coefficient of urate (MTACurate) was similar to the MTACNa+. The MTAC is the product of the elimination constant (ke) and the volume of distribution (VD), the mean intraperitoneal volume. Because VD is known, the ke Na+ in each patient can be equated with the ke urate. The Na+ mass transfer from the circulation to the dialysate by diffusion can then be calculated for any time point during a dwell (Dt). Dt was subtracted from the measured Na+ dialysate concentration at 60 minutes. The corrected D/P Na+ then represents the actual Na+ sieving. Using 3.86% glucose dialysate, this approach was investigated in 15 stable peritoneal dialysis (PD) patients (normUF) and in 9 PD patients with low ultrafiltration (lowUF, < 400 mL/4 hours). The MTACurate was calculated according to Waniewski (W) and according to the Garred model (G). Similar calculations were also performed for the MTAC of creatinine (MTACcreat). Initial D/P Na+ was not different between the groups. When no diffusion correction was made, D/P60 Na+ in the lowUF group (median 0.898, range 0.870-0.949) was significantly higher (p < 0.025) than D/P60 Na+ in the normUF group (median 0.881, range 0.816-0.899). The difference disappeared after diffusion correction regardless of the correction model applied. However, at 240 minutes, D/P Na+ in the normUF group was significantly lower than in the lowUF group (median 0.880, range 0.839-0.952 vs median 0.942, range 0.866-0.987; p < 0.004). Even after correction, D/P Na+ in the normUF group was significantly lower: 0.847 normUF versus 0.893 lowUF (Wurate, p < 0.005); and 0.842 normUF versus 0.890 lowUF (Gcreat, p < 0.003). The correlation between the Wurate (the best theoretical diffusion correction) and Gcreat (the least) was: y = 0.99x + 0.0037. Furthermore, Bland and Altman analyses of Wurate and Gcreat at both 60 and 240 minutes resulted in random distribution around the means, with a slight overestimation in relation to the magnitude of Gcreat, as was expected. Gcreat can be used to make an accurate estimation of the contribution of Na+ diffusion in the time course of D/P Na+. It provides a simple way to more precisely determine Na+ sieving, and therefore TCWT. In conclusion, to avoid overestimation of impaired channel-mediated water transport, a Na+ diffusion correction should be made when D0 is not equal to P0 or in the case of a large vascular surface area. PMID- 10682075 TI - Prevalence of gastroesophageal reflux disease in peritoneal dialysis and hemodialysis patients. AB - The prevalence of gastrointestinal reflux disease (GERD) in hemodialysis (HD) and peritoneal dialysis (PD) patients was assessed at a single center with a self administered questionnaire previously used in a general population. It defines (GERD as the presence of heartburn or acid regurgitation, or both. Risk factors for GERD and GERD-associated symptoms were also evaluated. In the studied population, 29.7% of patients had frequent GERD (heartburn, acid regurgitation, or both symptoms weekly). Frequent GERD was reported by 44.7% of PD patients versus the 18.9% reported by HD patients and the 19.8% reported by the general population. PD and HD patients had similar GERD severity scores [2.3 +/- 0.7 vs 1.9 +/- 0.8, mean +/- standard deviation (SD)]. PD and HD patients reported atypical GERD symptoms at rates similar to those reported by the general population, but having GERD made some atypical GERD symptoms more likely (p < 0.05, Fisher's exact test). In a logistic model, age < 60 [odds ratio (OR) 5.6, confidence interval (CI) 1.5-21.3], smoking (OR 4.7, CI 1.3-16.9), and body mass index > or = 27 (OR 3.9, CI 1.2-13.0) predicted GERD. Sex, race, diabetes, PD, non steroidal anti-inflammatory drugs (NSAIDs), calcium channel blockers, and coffee and alcohol use did not. GERD is more common in PD patients than in HD patients or in the general population. It is not clear whether PD per se is a risk factor for GERD. PMID- 10682076 TI - Response to triple treatment with omeprazole, amoxicillin, and clarithromycin for Helicobacter pylori infections in continuous ambulatory peritoneal dialysis patients. AB - In this study, the response to triple treatment with omeprazole, amoxicillin, and clarithromycin was investigated in continuous ambulatory peritoneal dialysis (CAPD) patients with Helicobacter pylori (Hp) infections. The study enrolled 20 CAPD patients (11 male, 9 female) who had dyspeptic complaints. The mean age of the patients was 46 (range: 21-65). The study also enrolled, as a control group, 124 patients (66 male, 58 female) who had no systemic disease, but who had upper gastrointestinal endoscopy for dyspeptic complaints. The mean age of the patients in the control group was 47 years (range: 20-74 years). Upper gastrointestinal endoscopy, rapid urease test (CLO test), and direct histologic examination were carried out to detect Hp infection. Hp infection was detected in 10 cases (50%) in the CAPD group and in 53 cases (43%) in the control group. In both groups, patients with Hp infection received the triple treatment of omeprazole 20 mg twice daily for 30 days, amoxicillin 500 mg thrice daily for 15 days, and clarithromycin 500 mg thrice daily for 15 days. To assess response to treatment, upper gastrointestinal endoscopy, CLO test, and direct histologic examination were repeated 3 months after initiation of the treatment. Hp was eradicated in all of the 11 CAPD patients (100%), and in 42 of the control patients (92%). Our results suggest that the triple treatment with omeprazole, amoxicillin, and clarithromycin for Hp infection is as effective in CAPD patients as in the normal population. PMID- 10682077 TI - Tumor necrosis factor alpha as a uremic toxin: correlation with neuropathy, left ventricular hypertrophy, anemia, and hypertriglyceridemia in peritoneal dialysis patients. AB - Tumor necrosis factor alpha (TNF alpha) is usually excreted by the kidney. In dialysis patients, it accumulates. TNF alpha has been implicated in the pathogenesis of malnutrition, diabetic neuropathy, and erythropoietin resistance. We studied TNF alpha plasma levels in 49 stable peritoneal dialysis (PD) patients, with the aim of correlating those levels with the presence and severity of peripheral neuropathy, hypertrophic cardiomyopathy, and anemia. Kt/Vurea' residual renal creatinine clearance (CrC), nutritional markers, and general biochemistry were also determined. The average plasma level of TNF alpha was 67 +/- 32 pg/mL (range: 18.1-156.3 pg/mL; normal value 3-20 pg/mL). No correlation was observed between TNF alpha and KT/Vurea' but a negative correlation with CrC was seen (r: -0.37, p < 0.05). TNF alpha levels were higher in patients with neuropathy as compared to patients with normal results (72.5 +/- 32 pg/mL vs 44 +/- 22 pg/mL, p < 0.05). Patients with neuropathy also showed a lower CrC value (1.5 +/- 1.7 mL/min vs 3.9 +/- 2.6 mL/min, p < 0.01). TNF alpha levels were higher in patients with left ventricular hypertrophy (LVH) with respect to normal individuals (70.4 +/- 32 pg/mL vs 38.5 +/- 20.8 pg/mL, p < 0.05). Patients with severe LVH showed the lowest CrC value. A direct, significant relationship was found between TNF alpha levels and weekly erythropoietin dose (r: 0.41, p < 0.05). Patients with hypertriglyceridemia or taking lipid-lowering agents showed a positive linear correlation between TNF alpha and triglycerides (r = 0.7, n = 14, p < 0.05). These data suggest that accumulation of TNF alpha may contribute to the development or maintenance of some neurologic, hematologic, and cardiac complications of uremic syndrome. Loss of residual renal function conditions an increment in TNF alpha levels. These data continue to add support to the idea that TNF alpha may be considered a uremic toxin. PMID- 10682078 TI - Predictors of survival in continuous ambulatory peritoneal dialysis patients: the importance of left ventricular hypertrophy and diabetic nephropathy. AB - We retrospectively evaluated the factors that are prognostic in long-term continuous ambulatory peritoneal dialysis (CAPD). From 1986 to 1997, 91 CAPD patients (59 male, 32 female, mean age 48 years) entered the study. Their primary renal diseases were chronic glomerulonephritis (CGN, n = 80), diabetic nephropathy (DN, n = 10), and polycystic kidney disease (PKD, n = 1). The roles of primary renal disease, hypertension, left ventricular hypertrophy (LVH), left ventricular ejection fraction (LVEF), cardiac sympathetic activity, anemia, hypoalbuminemia, and plasma concentration of parathyroid hormone (PTH) on patient prognosis were analyzed. Among the 91 CAPD patients, 26 died during the observation period. Of these deaths, 17 resulted from cardiovascular diseases including cerebrovascular events (n = 7), myocardial infarction (n = 2), sudden death (n = 7), and aortic aneurysmal rupture (n = 1). Nine patients died of non cardiovascular events. Sclerosing encapsulating peritonitis and others, mainly cachexia, accounted for 2 and 7 of these deaths, respectively. The 5-year survival rate was 74%; the 10-year rate was 49%. The cumulative 5- and 10-year success rates of CAPD were 69% and 39%, respectively. DN, hypertension, severe LVH (more than 200 g/m2), and hypoalbuminemia were contributors to poor prognosis. Among these, DN and severe LVH were the two main predictors by Cox proportional hazards model. We conclude that CAPD patients with DN or severe LVH, or both, have a greater chance of drop-out from cardiovascular events. PMID- 10682079 TI - Peritoneal dialysis prescriptions for diabetic patients. AB - Peritoneal dialysis offers several advantages for the treatment of diabetic patients with renal failure. The diabetic patients is often affected by comorbid conditions that influence the dialytic prescription and the clinical outcome. This article reviews the pertinent pathophysiology present in the diabetic patient with advanced renal insufficiency in an attempt to make specific recommendations for the initiation of peritoneal dialysis and the formulation of an adequate prescription. PMID- 10682080 TI - Peritoneal dialysis: matching the prescription to the membrane. AB - Peritoneal transport characteristics determine solute clearance and ultrafiltration and influence nutrition; matching the prescription to the membrane type is therefore important. The prognostic implications of peritoneal transport rate are here reviewed with emphasis on high transport states. The minimal adequate dose is discussed in the light of three important issues: (1) the relative importance of the urea and creatinine kinetic parameters; (2) the potential contributions of renal and peritoneal clearance to outcome; and (3) the influence of peritoneal transport rates on minimum dose. Specific recommendations are made for the formulation of prescriptions according to membrane type. PMID- 10682081 TI - Nonsurgical implantation of Tenckhoff peritoneal catheters in patients on continuous ambulatory peritoneal dialysis. AB - From 1994 to 1998, 54 Tenckhoff peritoneal catheters were implanted in 49 patients undergoing continuous ambulatory peritoneal dialysis. The implantation technique included bedside insertion of peritoneal catheters via trocar under local anesthesia. Early and late complications of this technique were comparable with surgical techniques. Cumulative survival of all catheters was 91% after one year, 78% after two years, 71% after three years, and 68% after four years. Our results suggest that the percutaneous technique for insertion of peritoneal catheters remains an easy, safe, and useful technique in the management of end stage renal disease patients. PMID- 10682082 TI - Long-term, successful peritoneal dialysis: end-stage renal disease core indicators study data. AB - The proportion of U.S. dialysis patients using peritoneal dialysis (PD) continues to fall. The reasons for the decline are presumably related to reduced patient recruitment and poor patient retention (technique failure). Yet, the 1998 Peritoneal Dialysis Core Indicators Study (PD-CIS) report suggests that PD provides satisfactory "long-term" renal replacement for many patients, and, further, that this capacity is becoming more obvious and is beginning to mimic center hemodialysis results. Several patient characteristics suggest that the key to successful "long-term" PD involves the delivery of "adequate dialysis" and that increasing attention to PD prescription and dialysis delivery has been accompanied by an improvement in PD technique success. PMID- 10682083 TI - Peritoneal dialysis immediately post transplantation. AB - Many peritoneal dialysis (PD) patients undergo successful renal transplantation. The outcome is similar to that for transplanted hemodialysis (HD) patients. For delayed graft function, dialysis is necessary in the immediate post-transplant period. PD can be undertaken, with a small risk of peritonitis and other PD related complications. Delayed graft function is less prevalent in patients who were on PD before transplantation than in those who were on HD. The dialysis modality may have an impact on graft survival. The graft thrombosis rate is unaffected by the prior mode of dialysis. Catheter-related problems are infrequent, but there must be a low threshold for catheter removal for any sign of non response to therapy for peritonitis or for exit-site and tunnel infections. PD can be undertaken successfully in the immediate post-transplant period. PMID- 10682084 TI - Differences in assessment of patients with satisfactory or complicated continuous ambulatory peritoneal dialysis courses. AB - The aim of our study was a comparison of comorbid scores, peritonitis rates, dialysis adequacy, and nutritional parameters in continuous ambulatory peritoneal dialysis (CAPD) patients. Patients were separated into two groups: those who, in the course of CAPD, were ingood clinical condition and underwent renal transplantation (group I, n = 11), and those who had to discontinue CAPD treatment (group II, n = 16) owing to death caused by comorbid disease or owing to transfer to hemodialysis for technique failure related mainly to recurrent peritonitis. Clinical scores were lower in group II, showing significantly more insomnia, weakness, and anorexia. The PET D/P creatinine, mean adequacy parameter, and urine output were similar in groups I and II. Daily protein intake (DPI) and daily energy intake (DEI) showed higher values in group I than in group II when expressed in g/kg and kcal/kg total body mass (TBM) respectively (DPI 1.09 +/- 0.15 g/kg TBM vs 0.92 +/- 0.31 g/kg TBM, p = 0.036; DEI 36.3 +/- 4.3 kcal/kg TBM vs 31.0 +/- 9.0 kcal/kg TBM, p = 0.048), but the intakes were not significantly different when calculated per kilogram ideal body mass (IBM). Lean body mass as a percent of total mass was 77.7% +/- 7.8% versus 73.9% +/- 6.8% (p = 0.048) in groups I and II respectively. Group I showed lower serum cholesterol than group II (179 +/- 33 mg/dL vs 231 +/- 41 mg/dL, p = 0.001) despite higher dietary intake of cholesterol (367 +/- 137 mg/day vs 251 +/- 97 mg/day, p = 0.016), correlating with DPI (r = +0.673, p = 0.023). Our results indicate that under conditions of similar CAPD adequacy, patients with a satisfactory course of CAPD therapy have higher dietary intake and are better nourished than those with a poor outcome. The changes in nutrition seem to be related to comorbid diseases and complications of CAPD therapy. Increased cholesterol level, associated with a diminished DPI, is prognostic of a poor outcome for CAPD patients. PMID- 10682085 TI - Peritoneal dialysis reduces the use of non native fistula access in dialysis programs. AB - Access problems remain the major difficulty associated with chronic hemodialysis. Despite recent recommendations by the Dialysis Outcomes Quality Initiative (DOQI) that native arteriovenous (AV) fistulae are the optimal form of vascular access, grafts and central catheters are used by many patients. We analyzed our large Canadian regional dialysis program, which has a high prevalence of peritoneal dialysis, to examine the effect of dialysis modality choice on vascular access utilization. Point prevalence data were collected from our program in October 1997, and technique and patient survival data for the period 1990-1996 were analyzed and compared to data for the remainder of Canada from the Canadian Organ Replacement Register. Mortality rate ratios were estimated using a Poisson regression model to correct for comorbidity, age, and end-stage renal disease etiology. Of 141 in-center hemodialysis patients, 91 had an AV fistula, 1 had a polytetrafluoroethylene (PTFE) graft, and 49 were catheter-dependent. The program also included 20 home hemodialysis patients with AV fistulae, and 156 patients on peritoneal dialysis. No mortality risk differences between hemodialysis and peritoneal dialysis are seen in our center, nor are they seen for each modality in comparison with the remainder of Canada. Technique survival for peritoneal dialysis at our center was about 80% at 2 years, significantly greater than for Canada. For the program as a whole, 49% of patients used peritoneal dialysis 35% a native AV fistula, and 15% a central catheter. For Canada and the U.S.A. respectively, the comparable data were: peritoneal dialysis, 32% and 17%; native fistula, 33% and 15%; PTFE, 19% and 41%; and central catheter 16% and 27%. These data suggest that the use of peritoneal dialysis may allow reduced use of non native AV fistula access without mortality penalty. PMID- 10682086 TI - Tidal peritoneal dialysis to achieve comfort in chronic peritoneal dialysis patients. AB - Patients with end-stage renal disease on chronic peritoneal dialysis (CPD) can usually tolerate continuous ambulatory peritoneal dialysis (CAPD) or continuous cycling peritoneal dialysis (CCPD) without abdominal discomfort or pain. In some patients, pain or discomfort occurs with complete drain of the peritoneal dialysis solution or upon initiation of dialysis filling when the peritoneal cavity is empty. We report on the use of tidal peritoneal dialysis (TPD) as a modality to alleviate this pain. Of 136 patients in our CPD unit, 18 (13%) were complaining of pain with complete drain or upon instillation of PD fluid. All were placed on TPD after other causes for abdominal pain were excluded. Six patients were placed on 25% TPD, and 12 patients on 50% TPD. The mean Kt/V of the patients on TPD was 2.46 +/- 0.68. With TPD, all patients had complete relief of abdominal discomfort. Patients who develop abdominal pain with complete drain or fill when the abdominal cavity is empty would benefit from TPD and be able to continue with CPD. PMID- 10682087 TI - Hemodialysis together with peritoneal dialysis is one of the simplest ways to maintain adequacy in continuous ambulatory peritoneal dialysis. AB - When long-term peritoneal dialysis (PD) is performed without change in the dialysis prescription, uremic symptoms appear owing to insufficient dialysis dose. In such cases, an increase in dialysate volume is required, but this increase is difficult to obtain in all patients owing to limitations in abdominal volume, lifestyle, or body weight. A combination of PD and hemodialysis (HD) is the simplest method of overcoming these limitations. Combination therapy--HD once per week for 4 hours and PD 6 days per week--was performed in our patients. The total weekly dialysis dose (urea) was calculated as follows: to convert the dialysis dose by HD to that of continuous treatment, the equivalent renal urea clearance (EKR) was calculated and added to the dialysis dose by PD. Combination therapy was performed in 12 patients. The reasons for the combination therapy included ultrafiltration (UF) loss in 2 patients, uremic symptoms in 3 patients, poor fluid management in 5 patients, umbilical hernia in 1 patient, and hydrothorax in 1 patient. Total Kt/V per week was increased from 1.61 +/- 0.19 to 2.05 +/- 0.25 in these patients. In the 2 patients with UF loss, weight control became easier after the combination therapy was started, and this control was possible with hypotonic dialysate alone. In patients with uremic symptoms, the symptoms improved; furthermore, dermal pigmentation improved in these patients. In summary, the dialysis dose was increased and body fluids became controllable after inducing combination therapy, resulting in improvement uremic symptoms and increased quality of life. PMID- 10682088 TI - Estimation of residual glomerular filtration rate and renal Kt/Vurea from creatinine clearance in end-stage renal disease patients. The Netherlands Cooperative Study on the Adequacy of Dialysis. AB - Residual glomerular filtration rate (rGFR) and renal Kt/Vurea are important parameters in clinical practice and in cohort studies. The calculation of these parameters requires analysis of urea in a 24-hour urine collection and in a simultaneously obtained plasma sample. In clinical practice, urea clearance is not always determined, but creatinine clearance usually is. The aim of the present study was to assess how well rGFR and renal Kt/Vurea can be estimated from creatinine clearance in end-stage renal disease (ESRD) patients. Of new Dutch ESRD patients, 365 were consecutively included in this study at the start of their chronic dialysis treatment. The estimation models were based on a random sample of two-thirds of the patients; the models were validated on the remaining one-third. We built models for pre-dialysis and peritoneal dialysis (PD) patients together (pre + PD group), and separate models for hemodialysis (HD) patients. Mean measured rGFR of pre + PD patients in the validation group was 6.3 mL/minute. The limits of agreement (LoAs) between estimated and measured rGFR were within -1.5 and +1.5. Mean measured rGFR in HD patients was 3.1 mL/minute (LoAs: -0.3 and +0.3). These relatively small limits of agreement reveal that, should urea clearance be missing, rGFR can be estimated by a formula in which creatinine clearance and 24-hour urine production are included. The estimation of renal Kt/Vurea from creatinine clearance was less precise. PMID- 10682089 TI - Trends in initiation of dialysis in an urban dialysis clinic in the United States: a long way from dialysis outcomes quality initiative guidelines. AB - The guidelines issued for peritoneal dialysis by the National Kidney Foundation Dialysis Outcomes Quality Initiative (NKF-DOQI) suggest initiation of dialysis at a glomerular filtration rate (GFR) of 10.5 mL/min/1.73 m2. We undertook this study to determine trends at a center in Philadelphia. Using the MDRD (Modification of Diet in Renal Disease) formula, we estimated the GFR (mL/min/1.73 m2) at the time of first dialysis for patients starting on dialysis at the center between 1994 and 1996. Data on 72 of the 86 new patients were available. Of the 72, 69 patients (96%) were black and 29 (40%) were diabetic. The estimated GFR at the time of dialysis initiation was 6.3 +/- 0.3 mL/min/1.73 m2. Only 3 patients (4%) had a GFR > 10.5; 25 patients (35%) had GFR < 5.0. Patients with a lower GFR had more severe acidosis (HCO3 15.8 +/- 0.5 mmol/L vs 19.5 +/- 0.9 mmol/L, p = 0.0023) and greater impairment in divalent ion metabolism (PO4: 7.6 +/- 0.2 mg/dL vs 5.6 +/- 0.3 mg/dL, p < 0.0001). Diabetic patients were initiated on dialysis at a higher level of GFR than non diabetic patients (7.2 +/- 0.3 mL/min/1.73 m2 vs 5.7 +/- 0.4 mL/min/1.73 m2, p = 0.0087). Even though diabetic patients had higher GFR and lower serum creatinine (8.6 +/- 0.5 mg/dL vs 11.8 +/- 0.9 mg/dL, p = 0.0021) than non diabetic patients, blood urea nitrogen (BUN) was similar in the two groups (89 +/- 3.7 mg/dL vs 96.8 +/- 5.8 mg/dL, p = 0.73). This difference may indicate that nondiabetic patients had a greater decline in protein intake. Finally, a trend towards initiation of dialysis at higher levels of renal function was seen with time (GFRs 5.5 +/- 0.4 mL/min/1.73 m2, 6.4 +/- 0.4 mL/min/1.73 m2, and 6.9 +/- 0.6 mL/min/1.73 m2 for 1994, 1995, and 1996 respectively; p = 0.058, 1994 vs 1996). The trend was stronger for diabetic patients (GFR 6.3 +/- 0.4 mL/min/1.73 m2 and 8.1 +/- 0.7 mL/min/1.73 m2 for 1994 and 1996 respectively; p = 0.06) than for non diabetic patients (GFR 5.1 +/- 0.5 mL/min/1.73 m2 and 6.2 +/- 0.7 mL/min/1.73 m2 for 1994 and 1996 respectively; p = 0.22). We conclude that initiation of dialysis is delayed in urban centers, particularly in non diabetic patients, and that the current practice is far below the DOQI recommendations. PMID- 10682090 TI - The importance of an extra hour of cycler therapy to obtain better adequacy. AB - As continuous ambulatory peritoneal dialysis (CAPD) patients lose residual renal function, it frequently becomes impossible from them to obtain adequate dialysis unless the dialysis prescription is changed. Increasing the dwell volumes, increasing the frequency of exchanges, and using a night-exchange device or a cycler are the means available to improve adequacy. In an effort to obtain dialysis adequacy, we studied how an extra hour on cycler therapy can contribute to improving dialysis adequacy. Over 18 months, we optimized solute clearance using the PD Adequest program (Baxter Healthcare Corporation, Chicago, Illinois, U.S.A.) in 70 patients. After finding the best total cycler volume, we compared the weekly creatinine clearance and weekly Kt/V from 8-hour cycler therapy to that from 9-hour cycler therapy for the four types of membrane transport. Adding one extra hour on cycler therapy improved weekly creatinine clearance by 3-6.5 L and the weekly Kt/V by 0.16-0.20. When patients are marginally approaching the required weekly Kt/V or creatinine clearance, an extra hour on the cycler may help to achieve the desired adequacy. PMID- 10682091 TI - Bioelectrical impedance analysis in the evaluation of the nutritional status of continuous ambulatory peritoneal dialysis patients. AB - The authors evaluated the nutritional status of 47 continuous ambulatory peritoneal dialysis (CAPD) patients, 26 men (age 58.9 +/- 14.6 years, duration on CAPD 27.3 +/- 18.3 months) and 21 women (age 56.2 +/- 14.9 years, duration on CAPD 34.5 +/- 23.4 months), using subjective global assessment (SGA), an established method for the nutritional assessment of dialysis patients, and bioelectrical impedance analysis (BIA: body cell mass, fat mass, and phase angle). Of the studied patients, 19 were diabetic (age 59.7 +/- 13.8 years, duration on CAPD 29.2 +/- 19.7 months) and 28 were non diabetic (age 53.9 +/- 14.3 years, duration on CAPD 31.5 +/- 21.8 months. According to SGA, 24 patients were scored as well nourished (Group I), 18 as mildly malnourished (Group II), and 5 as moderately malnourished (Group III). No patient scored as severely malnourished (Group IV). Analysis of the main nutritional parameters for the subgroups revealed a proportional decrease in phase angle, with a statistically significant correlation (p < 0.009) between phase angle and SGA. No significant difference was seen in serum albumin levels between patients in Group I and Group II, but the mean level was significantly lower in patients in Group III compared to Group I. The nutritional and biochemical data in diabetic patients and non diabetic patients were not significantly different. BIA phase angle seems to be a simple and reliable method for the routine assessment of nutritional status in CAPD patients. PMID- 10682092 TI - Incidence of gingival hyperplasia caused by calcium antagonists in continuous ambulatory peritoneal dialysis patients. AB - Calcium antagonists are widely used for the treatment of cardiovascular diseases in patients receiving dialysis therapy. The incidence of gingival hyperplasia has been reported as 10%-20% in patients treated with calcium antagonists in the general population. However, precise reports examining the incidence or pathogenesis of gingival hyperplasia in continuous ambulatory peritoneal dialysis (CAPD) patients are lacking. We recruited 54 CAPD patients. Three patients treated with long-acting nifedipine and one patient treated with felodipine were reported by a periodontist to have gingival hyperplasia. No patients were taking amlodipine and other calcium antagonists. After discontinuation of calcium antagonists, gingival hyperplasia disappeared within 1 month. Based on these results, we suggest that it is important to examine whether the gingiva is overgrown in CAPD patients taking calcium antagonists. PMID- 10682093 TI - Evidence that gender difference affects peritoneal dialysis capacity in continuous ambulatory peritoneal dialysis patients. AB - The progression of renal disease is reported to be more rapid in male patients than in premenopausal females. However, few studies compare the difference in dialysis therapy between males and females. We compared the efficacy of peritoneal dialysis capacity, using measurements of retention volume and pelvic cavity by helical section of computed tomography (CT) in 6 male and 6 female patients. The patients did not differ significantly in age (males: 54 +/- 3 years; female: 56 +/- 4 years). Males were heavier than females (p < 0.05). Retention volume in the visceral cavity was significantly larger in males (1787 +/- 43 mL) than in females (1580 +/- 59 mL) (p < 0.05). Peritoneal dialysis capacity was evaluated by peritoneal equilibration test (PET). Although no significant differences were observed in the PET data, when the PET results at 4 hours were divided by body weight in kilograms, a significant difference between males and females was seen (p < 0.05). There was a mild, but not significant, correlation between the volume of the pelvis as measured by helical CT and the PET data per kilogram body weight (p = 0.07). These results suggest that gender differences in peritoneal dialysis capacity relate partially to the difference in pelvic cavity volume. PMID- 10682094 TI - The use of peritoneal dialysis in special situations. AB - This brief review outlines several situations in which peritoneal dialysis (PD) can be used to address clinical situations that are out of the ordinary for end stage renal disease (ESRD). For example, PD methodology can be used not only to treat ESRD patients with difficult psychosocial problems that obviate other dialysis options, but also to control ascites accumulation in patients with liver failure, to treat congestive heart failure in azotemic patients with progressive cardiomyopathy, to administer systemic medication via the peritoneal cavity, and to provide additional clearance in demanding circumstances. In discussing these unusual applications for PD, we open the door to extending the indications for PD to a broader spectrum of clinical problems. PMID- 10682095 TI - Residual volume in continuous ambulatory peritoneal dialysis patients from various solute calculations. AB - Residual volume (RV) has been measured using various methods and various solutes. This prospective study set out to determine residual volume by using various solute calculations, and to estimate mean residual volume in stable Thai CAPD patients. Complete data from 9 patients (6 males, 3 females) were analyzed. Residual volume was calculated using various solutes including sodium (Na), potassium (P), glucose (G), urea (U), and creatinine (C). The calculation using sodium was too variable to use to calculate a mean. Mean of residual volume was calculated from the other solutes as RV1 (mean of PUC), RV2 (mean of GUC), and RV3 (mean of PGUC). Mean residual volumes did not correlate with sex, body weight, body surface area, hematocrit, albumin, or membrane characteristics (drained volume). Comparing all means calculated in our study, creatinine was the solute that yielded the most accurate mean, with a correlation coefficient of 0.877, 0.965, and 0.956 for RV1, RV2, and RV3 respectively. In this study, mean residual volumes were 212.49 +/- 100.03 mL, 255.23 +/- 142.58 mL, and 250.42 +/- 121.60 mL for RV1, RV2, and RV3 respectively. PMID- 10682096 TI - Failure of icodextrin to provide adequate ultrafiltration in continuous ambulatory peritoneal dialysis patients. AB - Icodextrin, a starch-derived glucose polymer with an average molecular weight of 20,000 D, has been developed partly as a response to some of the disadvantages of dextrose. It has been suggested that icodextrin solutions are able to provide sustained ultrafiltration (UF) over long dwell times of 8-12 hours in continuous ambulatory peritoneal dialysis (CAPD). In this paper we describe three patients on CAPD: 2 males and 1 female aged 60, 67, and 58 years respectively, duration on CAPD 47, 60, and 15 months respectively. All of these patients, who were categorized as high transporters according to peritoneal equilibration test (PET) results, presented early signs of ultrafiltration loss with no evidence of peritoneal inflammation. Icodextrin solution was used in a single nightly exchange with 10-12 hours' dwell, for a period of 5-30 days. In all of these cases, icodextrin solution failed to provide adequate ultrafiltration and the patients returned to the previously used regime of five daily hypertonic exchanges of 3.86% glucose concentration. Although these negative results were not clearly explained, we report these three cases because they exemplify some limitations of icodextrin solution to provide adequate ultrafiltration, at least in a small number of CAPD patients. PMID- 10682097 TI - Incremental initiation of peritoneal dialysis: current practice. AB - The Dialysis Outcomes Quality Initiative (DOQI) guidelines define initiation of peritoneal dialysis (PD) as "timely" when dialysis is started using deteriorating renal function and nutritional indices as criteria--before the appearance of frank uremia--and as "incremental" when the initial dose of PD is small, bringing total clearances to or above target values, with the understanding that the dose of PD will be increased as renal function is lost. Current practice regarding timely/incremental PD initiation was studied by analyzing the responses to a questionnaire distributed to 250 U.S. and Canadian nephrologists. A total of 89 responses were received. Only 30 responders (33.7%) practice incremental PD initiation. The more frequent reasons for not practicing incremental PD initiation included anticipated patient noncompliance, the desire to obtain the highest possible clearances, disagreement with the DOQI rationale, and late referral of patients to the nephrologists. Renal clearances were used by 58 responders (65.2%) as independent criteria to start dialysis. Creatinine clearance, at a cut-off level of 9.9 +/- 2.1 mL/minute, was the criterion most frequently used to start PD. Nutritional indices were used by 66 responders (74.2%) as criteria to start PD. Frequently used nutritional indices were serum albumin, body weight, normalized equivalent of protein nitrogen appearance (nPNA), and dietary assessment. The survey provides clues about research studies and educational activities that will be needed in the future. PMID- 10682098 TI - Small-solute clearances in diabetic subjects on continuous ambulatory peritoneal dialysis: comparison to nondiabetic subjects. AB - Normalized clearances for urea and creatinine were compared between 121 diabetic subjects (256 clearances) and 181 nondiabetic subjects (357 clearances) on continuous ambulatory peritoneal dialysis (CAPD) with four 2-L exchanges daily. Urea clearance was normalized by VWatson (Kt/Vur), while creatinine clearance was normalized by both VWatson (Kt/Vcr) and body surface area (Ccr). Height, weight, body water (V), and body surface area did not differ between the diabetic and the nondiabetic groups. Also, renal Kt/Vur, renal Kt/Vcr, renal Ccr, and peritoneal Kt/Vur did not differ between the groups. Weekly peritoneal Kt/Vcr (diabetic group 1.36 +/- 0.38, nondiabetic group 1.31 +/- 0.31, p = 0.048) and weekly peritoneal Ccr (diabetic group 47.6 +/- 11.0 L/1.73 m2, nondiabetic group 45.4 +/ 9.2 L/1.73 m2, p = 0.012) were both higher in diabetic subjects. The percentage of high/high-average transporters was higher in the diabetic group (64.9% vs 48.6% in nondiabetic group, p = 0.006). The following total (peritoneal + renal) weekly clearances were obtained: Kt/Vur, diabetic group 2.07 +/- 0.63, nondiabetic group 2.02 +/- 0.56, NS; Kt/Vcr, diabetic group 2.06 +/- 0.78, nondiabetic group 1.92 +/- 0.74, p = 0.026; Ccr, diabetic group 72.7 +/- 28.5 L/1.73 m2, nondiabetic group 67.2 +/- 26.4 L/1.73 m2, p = 0.013. Normalized total creatinine clearances are higher in diabetic subjects than nondiabetic subjects on the same CAPD schedule and with the same renal clearances of urea and creatinine and the same total Kt/Vur, because peritoneal creatinine clearances are higher in the diabetic subjects. This finding is caused by higher peritoneal transport in the diabetic subjects and is not an artifact caused by the normalization process. PMID- 10682099 TI - Peritoneal sclerosis--an overview. AB - The term peritoneal sclerosis encompasses a vast range of peritoneal alterations, from the low clinical impact manifestations associated with chronic peritoneal dialysis, to dramatic thickening of the peritoneal membrane, which is rare, but often life-threatening. The frequency, pathology, etiopathogenesis, clinical manifestations, diagnostic criteria, therapy, and prevention of peritoneal sclerosis are reviewed. Preliminary observations from the Italian Registry of Peritoneal Sclerosis, established in the framework of a program of the Italian Society of Nephrology, are reported. PMID- 10682100 TI - Comparing peritonitis in continuous ambulatory peritoneal dialysis patients versus automated peritoneal dialysis patients. AB - The purpose of our study was to compare the incidence of peritonitis between continuous ambulatory peritoneal dialysis (CAPD) treatment (Group I) and automated peritoneal dialysis (APD) treatment (Group II) taking into account the same population. We compared 20 patients with a follow-up of 215 patient-months on CAPD and 252 patient-months on APD. Demographic data, diagnosis, peritoneal equilibration test (PET) results, adequacy, and peritonitis rate were analyzed. Diagnoses included glomerulopathy 35%, autosomal dominant polycystic kidney disease (ADPKD) 20%, Type II diabetes 10%, systemic lupus erythematosus 5%, interstitial nephritis 5%, nephrolitiasis 5%, and unknown 20%. PET results showed that the group consisted of 30% high transporters, 45% high-average transporters, and 25% low-average transporters. Kt/V for Group I was 1.3 +/- 0.3, and for Group II, 1.83 +/- 0.48. Creatinine clearance for Group I was 43.64 +/- 7.31 L/week/1.73 m2, and for Group II, 52.42 +/- 13.47 L/week/1.73 m2. Group I presented a peritonitis rate of 8.3 episodes/patient-month, and Group II presented a rate of 18.9 episodes/patient-month. Gram-positive organisms were responsible for 49.8% of episodes of peritonitis in Group I (S. aureus 26.6%, S. epidermidis 16.6%, others 10%) and 83% of peritonitis episodes in Group II (S. epidermidis 46.6%, S. aureus 20%). Gram-negative organisms were responsible for 16.5% of episodes of peritonitis in Group I. No gram-negative peritonitis was seen in Group II. APD patients developed two cases of candida peritonitis. Our preliminary results show that Group II exhibited a decrease in peritonitis rate while achieving better adequacy. In CAPD and APD peritonitis, gram-positive organisms predominated. In APD, we observed an increase in S. epidermidis incidence. No gram-negative organisms were observed in APD. It seems that APD is a safer treatment owing to the lower peritonitis incidence. PMID- 10682101 TI - Peritoneal dialysis--associated peritonitis caused by gram-negative bacteria: characteristics similar to spontaneous bacterial peritonitis? AB - The aim of the study was to investigate the characteristics of PD-related peritonitis caused by gram-negative bacteria (GNP). We retrospectively studied the medical records of 164 patients (114 males, 50 females; mean age 46 +/- 15 years) who continued PD beyond 5 months between 1984 and 1998. The average observation time was 40 +/- 28 months (total of 6609 patient-months). A total of 166 episodes of peritonitis occurred during that time (mean incidence: 1 episode/40 patient-months). Of these, 35 were GNPs, and GNP incidence stayed almost constant over time. Most GNP patients (63%) recovered without complication with an average of 14 days' antibiotic treatment. In only 4 cases was PD abandoned. Clinical features of GNP were similar to those of spontaneous bacterial peritonitis (SBP). The unchanged incidence of GNP over time with advanced connection devices suggests that there are important mechanisms promoting micro-organisms of endogenous origin into the peritoneal cavity in PD patients. PMID- 10682102 TI - Exit-site and catheter care: review of important issues. AB - This brief review addresses the impact that several important aspects of catheter technology and exit-site care have on catheter-related infections and catheter longevity. The discussion includes exit-site microbiology, catheter configuration, exit-site care, and catheter salvage, following which a summary of recommendations is presented. PMID- 10682103 TI - Simultaneous catheter removal and replacement in peritoneal dialysis infections: update and current recommendations. AB - Problematic peritoneal dialysis infection is a major cause of catheter loss and interruption of peritoneal dialysis (PD) therapy. In selected instances, problematic infection can be successfully treated by removing and replacing the catheter while continuing with PD. Accumulated experience has helped to define the circumstances under which a removal/replacement procedure is likely to be safe and under which complications are likely to arise. It appears that simultaneous removal and replacement can be expected to succeed when problematic infection is associated with tunnel infection, with recurring peritonitis repetitively culturing the same organism but clearing between episodes, and with gram-positive organisms. Success is less likely in the presence of ongoing inflammation, of active infection, of gram-negative or fungal organisms, or of any evidence of intra-abdominal adhesions. We review the literature on which these criteria are based and conclude with updated recommendations. PMID- 10682104 TI - Outcome of Pseudomonas aeruginosa exit-site and tunnel infections: a single center's experience. AB - We reviewed the course and outcome of all Pseudomonas aeruginosa (PA) exit-site and tunnel infections (ESI/TI) that occurred at our home peritoneal dialysis (PD) unit over a 3-year period (July 1995 to June 1998). We documented PA ESI/TI in 19 out of a total of 467 patients. Of the 19 patients, 12 having local redness and tenderness but no discharge were treated conservatively with increased frequency of dressing with or without hydrogen peroxide locally. Of the 12 cases receiving local care, 7 resolved without recurrence over 14.4 months follow-up, while the remaining 5 developed persistent ESI/TI with discharge and required treatment with antibiotics. Seven more patients who initially presented with purulent discharge also received systemic antibiotics. Only 1 of the 12 patients with PA ESI/TI treated with antibiotics resolved; the remaining 11 patients developed PA peritonitis over a 1-month to 7-month period after the initial PA ESI/TI. In 2 of these 11 patients, simultaneous PD catheter removal and replacement was attempted for the treatment of PA ESI/TI, but these patients also developed PA peritonitis 1-3 weeks after the procedure. Of the 11 patients with PA peritonitis associated with PA ESI/TI, 1 died, 6 were transferred to permanent hemodialysis, and just 4 continued PD after PD catheter replacement. Though not frequent, PA ESI/TI is still a serious complication of home PD at our unit, resulting in ESI/TI-related PA peritonitis and catheter loss in 58% of cases. Local treatment of mild PA ESI/TI (redness and induration) seems to be effective. On the other hand, patients with purulent discharge are likely to develop peritonitis and technique failure despite antibiotic therapy. Early catheter replacement can be considered in these cases. PMID- 10682105 TI - Once-daily intraperitoneal cefazolin and oral ciprofloxacin as empiric therapy for the treatment of peritonitis. AB - The Ad Hoc Advisory Committee on Peritonitis Management has recommended intraperitoneal (i.p.) cefazolin and an aminoglycoside as empiric therapy for the treatment of peritonitis. Because most of our patients are on continuous cycler therapy, we developed a strategy of once-daily i.p. cefazolin and oral ciprofloxacin as empiric therapy. All patients in our unit that developed peritonitis were given a once-daily 2 g load of i.p. cefazolin, plus 500 mg ciprofloxacin orally, twice daily. Ciprofloxacin was given two hours after any phosphate binder or iron supplement. The i.p. cefazolin was allowed to dwell for at least six hours. The dialysate was then drained and chronic peritoneal dialysis (CPD) resumed. Organisms sensitive to cefazolin were treated for 14 days with once-daily cefazolin alone; resistant organisms were treated with alternative antibiotics. A total of 40 patients were treated with this empiric regimen. Of these, 35 (88%) successfully continued CPD therapy, 1 (2%) transferred to hemodialysis, and 4 (10%) expired two weeks after the onset of peritonitis. A total of 22 patients (55%) were treated successfully with once daily i.p. cefazolin. Although once-daily i.p. cefazolin and oral ciprofloxacin permitted continuation of CPD therapy in most patients in this study, the therapy was not optimal. While vancomycin may have provided better coverage, recent reports of vancomycin-resistant enterococci and Staphylococcus aureus present a major concern. PMID- 10682106 TI - Peritoneal toxicities of hypertonic dextrose dialysate. AB - This report reviews the peritoneal toxicities of hypertonic (4.25%) dextrose dialysate. In vitro incubation of peritoneal cell lines (polymorphonuclear cells, macrophages) in hypertonic dextrose solutions have demonstrated direct cytotoxicity, decreased phagocytosis and bacterial killing, and decreased production of superoxide and inflammatory cytokines and leukotrienes. Hypertonic dextrose dialysate also inhibits the proliferation of mesothelial cells and causes increased mesothelial production of transforming growth factor, potentially leading to peritoneal fibrosis. In vivo animal studies have also shown peritoneal toxicity of hypertonic dextrose dialysate. Recently, the use of peritoneal dialysate with high glucose concentration has been found to lead to the deposition of advanced glycosylation end-products in the peritoneal structures and to the development of high peritoneal solute transport. Fluid control in peritoneal dialysis should not be based on the liberal use of hypertonic dextrose dialysate. Instead, it should be achieved by restriction of salt and water intake and prescription of the peritoneal dialysis schedule (particularly the dwell time) based on peritoneal equilibration test (PET) findings. Newer osmotic agents may obviate the use of hypertonic dialysate in the future. PMID- 10682107 TI - Reliability of the 7-point subjective global assessment scale in assessing nutritional status of dialysis patients. AB - Subjective global assessment (SGA) is a method to score nutritional status in a standardized way. The original 3-point scale has been replaced by a 7-point scale. The reliability of the latter scale has never been tested. We therefore assessed inter-observer and intra-observer reliability. Furthermore, we examined the relationship of SGA with other objective nutritional parameters. In 13 hemodialysis and 9 peritoneal dialysis patients, two nurses assessed SGA. They re examined the same patients two weeks later. Anthropometric measurements and blood samples were taken at the first assessment. According to SGA, 2 patients (9%) were classified as severely malnourished, 6 (27%) as mildly malnourished, and 14 (64%) as well nourished. The 7-point SGA scale showed fair inter-observer reliability [intraclass correlation (ICC) = 0.72] and good intra-observer reliability (ICC = 0.88). A strong correlation was present between the 7-point SGA scale and body mass index (BMI) (r = 0.79, p < 0.001), % fat (r = 0.77, p < 0.001), and mid arm circumference (r = 0.71, p < 0.001). Lower correlations were found with mid arm muscle circumference and serum albumin. With respect to biochemical markers, the strongest relationship was found with prealbumin (r = 0.60, p = 0.004). We conclude that the 7-point SGA scale is a valid and reliable tool to assess nutritional status among end-stage renal disease patients. We suggest that one observer or a select group of observers perform the assessments to gain maximum benefit from the reliability of the SGA instrument. PMID- 10682108 TI - Phthalic acid is the main metabolite of the plasticizer di(2-ethylhexyl) phthalate in peritoneal dialysis patients. AB - Di(2-ethylhexyl) phthalate (DEHP) is the most commonly used plasticizer in polyvinyl chloride (PVC) plastics, and is therefore a major constituent of continuous ambulatory peritoneal dialysis (CAPD) bags. Because DEHP is not chemically bound, it leaches out of the plastic matrix. Recently, we found that leukocyte function in vitro is impaired by a mixture of metabolites of DEHP. In the present study, we investigated the metabolism of DEHP in patients on CAPD. The study group consisted of 10 stable patients, on CAPD for at least 6 months, using a plasticizer-containing PVC PD system [ANDY Plus (Fresenius Medical Care, Bad Homburg, Germany)]. Effluent dialysate and urine samples were collected over 24 hours, and a 10 mL blood sample was drawn. Concentrations of DEHP and its metabolites mono(2-ethylhexyl) phthalate (MEHP), phthalic acid (PA), and 2 ethylhexanol (2-EH) were determined in urine, dialysate, and serum using gas chromatography/mass spectrometry. Additionally, the degree of glucuronidation of the phthalic acid esters in urine were determined. In serum, dialysate, and urine, PA was the predominant metabolite of DEHP (0.205 +/- 0.067 mg/L, 0.284 +/- 0.180 mg/L, and 1.34 +/- 1.00 mg/L, respectively), but concentrations of MEHP were low (0.0100 +/- 0.0056 mg/L, 0.022 +/- 0.008 mg/L, 0.011 +/- 0.0064 mg/L, respectively). Urinary MEHP was glucuronidated to approximately 15%. PA was 35% eliminated as a glucuronide. Unlike healthy subjects, PD patients do not eliminate DEHP mainly in the form of MEHP or MEHP metabolites. They further break these compounds down to PA. The fact that concentrations of PA in urine exceed by far the respective serum concentrations indicates that PA is secreted by the kidney. Further research on the toxicological aspects of plasticizers in uremic patients should take these findings into account. PMID- 10682109 TI - Lack of interference of icodextrin on creatinine measurements. AB - Glucose has been reported to interfere in the analysis of creatinine by the Jaffe method. The potential interference of icodextrin and its primary metabolites (maltose, maltotriose, maltotetraose) on creatinine measurements has not previously been addressed. We evaluated the potential interference of icodextrin and its metabolites at various concentrations using both the Jaffe and Creatinine Plus methods. Interference was determined in samples containing 0.6-20 mg/dL creatinine in saline solution or in plasma (n = 6), and in dialysate samples (n = 6) spiked with icodextrin, maltose, maltotriose, and maltotetraose at concentrations up to twofold the level found in plasma and dialysate from patients treated using icodextrin. Results confirm that no interference occurs when using either the colorimetric Jaffe method or the enzymatic Creatinine Plus method at levels up to 65 g/L icodextrin, 2 g/L maltose, 2 g/L maltotriose, and 1 g/L maltotetraose, levels representing worst-case clinical concentrations. In addition, our results confirm that comparable values can be obtained using either the Jaffe or the Creatinine Plus method for the analysis of creatinine in uremic plasma and in dialysate samples. PMID- 10682110 TI - Low concentrations of glucose degradation products in peritoneal dialysis fluids and their impact on biocompatibility parameters: prospective cross-over study with a three-compartment bag. AB - The side effects of glucose degradation products (GDPs) in conventional peritoneal dialysis (PD) fluids are well described. Using the three-compartment bag concept--that is, in situ preparation of concentrated glucose solution into a standard ionic solution--a GDP-free solution can be processed. To investigate the possible impact of this product on biological and clinical parameters, we carried out a prospective cross-over study with 31 patients, comparing the short-term effects of conventional PD and GDP-free PD solutions. Classical peritoneal parameters and ultrafiltration rate did not change during the study. After three months and after six months with the three-compartment bag, cancer antigen 125 (CA125) concentration in overnight fluid increased significantly (p < 0.001) from 24.4 IU/mL to 44.4 IU/mL and 41.1 IU/mL respectively. CA125 decreased significantly (p < 0.01) to 21.7 IU/mL after three months with the conventional solution. No change in hyaluronan concentration was observed. A slight increase of procollagen III N-terminal peptide in overnight effluent with the GDP-free solution was followed by a significant reduction after three months with standard solution. In summary, our data show that the GDP-free PD fluid improves mesothelial cell mass and turnover even after a short-term period of three months. A better quality of PD solution is obtained by using the three compartment bag. PMID- 10682111 TI - Influence of apolipoprotein E genotype on lipid and lipoprotein levels in continuous ambulatory peritoneal dialysis patients. AB - Apolipoprotein (Apo) E has an important role in triglyceride (TG)-rich lipoprotein metabolism, and the genotype of Apo E is associated with premature coronary artery disease. Patients undergoing continuous ambulatory peritoneal dialysis (CAPD) develop various abnormalities of lipid metabolism and are prone to develop accelerated atherosclerosis. To investigate the distribution of Apo E genotype, and to evaluate the influence of Apo E polymorphism on lipid metabolism in CAPD patients, we measured Apo E genotypes, serum lipid, and lipoprotein levels in 54 CAPD patients (M:F = 1:1). The most common genotype of Apo E in the CAPD patients was E 3/3, found in 68.5%. The frequencies of the other genotypes were E 2/3, found in 14.8%, and E 4/3, found in 16.7%. No genotypic differences in Apo E were seen in the patients with regard to the presence of diabetes, the level of parathyroid hormone, or the transport characteristics of the peritoneal membrane. When comparing lipid levels by Apo E genotype, the total cholesterol and TG levels of E 2/3 patients were significantly higher than those of E 3/3 or E 4/3 patients. The differences in high-density or low-density lipoprotein cholesterol levels by Apo E genotype were not significant. In comparing lipoprotein levels by Apo E genotype, the Apo B and lipoprotein (a) levels of E 2/3 patients were significantly lower than those of E 3/3 or E 4/3 patients. In conclusion, the Apo E 3/3 genotype seems to be the most common genotype in CAPD patients, and the Apo E 2/3 genotype appears to be associated with high cholesterol and TG levels. These results demonstrate the need for further prospective studies in these subjects aimed at elucidating the impact of genetic variation at the Apo E locus on the development of atherosclerosis. PMID- 10682112 TI - Do dialysis patients need extra folate supplementation? AB - To assess folate status and to evaluate the need for conventional folate supplementation in patients on dialysis, we measured serum folate, vitamin B12, and red cell folate concentrations by radioimmunoassay. Thirty-four continuous ambulatory peritoneal dialysis (CAPD) patients and 60 hemodialysis (HD) patients who had not been supplemented with folate were enrolled. Serum folate levels (5.8 +/- 3.6 ng/mL vs 2.0 +/- 1.1 ng/mL, p < 0.001) and vitamin B12 levels (831.4 +/- 416.9 pg/mL vs 513.9 +/- 213.3 pg/mL, p < 0.001) were significantly higher in CAPD patients than HD patients. The red cell folate levels (849.7 +/- 489.4 ng/mL vs 491.0 +/- 253.2 ng/mL, p < 0.001) were also significantly higher in CAPD patients. The incidences of folate deficiency in CAPD and HD patients were overestimated using the cut-off value for serum folate concentration (3.0% vs 71.7%, respectively), but the incidence of true folate deficiency was lower using the cut-off value for red cell folate level (0.0% vs 10.0%, respectively). In conclusion, the true incidence of folate deficiency in stable CAPD and HD patients is surprisingly low, even in patients who may not be taking folate supplements. The need for conventional folate supplementation in patients with end-stage renal disease on dialysis must therefore be re-evaluated. Before the decision is made to use folate supplementation, measurement of red cell folate is essential to assess of folate reserves of the patients on dialysis. PMID- 10682113 TI - Peritoneal mesothelial cells produce complement factors and express CD59 that inhibits C5b-9-mediated cell lysis. AB - The CD59 membrane protein confers protection from C5b-9-mediated cell lysis. Because evidence exists for complement (C) activation and generation of C5b-9 in the peritoneal cavity during chronic peritoneal dialysis (CPD), we investigated, on mesothelial cell (MC) lines, the expression of CD59 and the production of C components. Four MC lines were obtained from children on CPD, and two from non uremic children. CD59 expression on MCs was investigated with anti-CD59 monoclonal antibody (mAb) and polyclonal goat immunoglobulin G (IgG). MC lines were positive for staining with anti-CD59 mAb. Western blotting analysis of MC membrane demonstrated a band with the same molecular weight as CD59. Incubation of MC with anti-CD59 mAb abrogated the protective effect of CD59 (100% cytotoxicity). C3, C4, and C6 were detected in the supernatants of MC; in non uremic MC supernatants, C5, C7, C8, and C9 were also detectable, and C4 concentration was tenfold higher. CD59 expression confers to MCs protection from C5b-9-mediated lysis. MCs produce C factors. These findings suggest that production of complement components and expression of CD59 on MCs could play a role both in peritoneal cavity infection (decreased complement production) and in peritoneal membrane damage (decreased CD59 expression and reduced remesothelialization owing to MC lysis). PMID- 10682114 TI - Five years' experience of a hospital-based home-care renal nursing service. AB - A pediatric home-care renal nursing service was initiated in our unit five years ago to provide direct respite care in the home for families on the dialysis and transplant program. This nursing post responded to parental requests for more practical support in the home. A trained children's nurse with renal and community nursing qualifications was recruited with charitable support. Between October 1993 and October 1998, 286 respite-care visits were performed. Of the visits, 57% were for children on continuous cycling peritoneal dialysis (CCPD), 20% were for pre-dialysis support (mainly supplementary feeding of children < 5 years old), 15% were for children on hemodialysis, and 8% were for children in the post transplantation period. The age of children receiving respite care ranged from 2 months to 15 years. Distances traveled from the unit to the home ranged from 5-150 miles with visit times of 2-10 hours. Of all visits, 60% were performed during the day, and 60% involved sibling care. The parental response to home-care support resulted in the incorporation of the home-care nurse into the unit's nursing budget. The new nursing post has raised issues of the professional accountability of home-care nurses, of patient confidentiality, and of communication with multi-disciplinary team members. Reflecting upon our five-year experience of home-care nursing has encouraged us to further develop our home support program for families living at increasing distances from our unit. PMID- 10682115 TI - Evidence of central hypothyroidism in children on continuous ambulatory peritoneal dialysis. AB - This study investigated the effects of chronic peritoneal dialysis on thyroid function and thyroid volume of patients with chronic renal failure (CRF). We measured the levels of serum and dialysate thyroid hormones [total thyroxine (TT4), total triiodothyronine (TT3), free thyroxine (fT4), and free triiodothyronine (fT3)], thyrotropin (TSH), thyroglobulin (Tg), and thyroid volume in 10 children on chronic peritoneal dialysis [9 continuous ambulatory peritoneal dialysis (CAPD), 1 continuous cycling peritoneal dialysis (CCPD)] at baseline and after one year. Serum levels in patients were compared with those in age- and sex-matched healthy children and were scored as normal, low, or high. At the beginning of study, serum levels were low for TT3 in 1 patient, for fT3 in 8 patients, for fT4 in 3 patients, and for Tg in 1 patient; serum TSH was high in 1 patient. At the end of study, serum levels were low for TT3 in 2 patients, for TT4 in 2 patients, for fT3 in 9 patients, for fT4 in 4 patients, for TSH in 2 patients, and for Tg in 3 patients. At the start of the study, only TSH and Tg levels could be detected in peritoneal dialysate; other parameters could not be measured. One year later, levels of TSH had decreased in 6 patients and increased in 3 patients, and Tg had increased in 8 patients, compared with baseline levels. To determine the effect of CAPD, baseline results were compared with mean levels at the end of the study. Although the mean levels of all parameters, except Tg, had decreased after one year, only the decrease in serum TSH was statistically significant. On the other hand, only the levels of Tg increased significantly in peritoneal dialysate. The mean value of thyroid volume also decreased after a year, but all values were within the normal range, and the decrease was not significant. No correlation was found between dialysis duration and any parameter after one year. In conclusion, we found a decrease in serum thyroid hormones, thyroid volume, and TSH in chronic peritoneal dialysis patients. We suggest that the low TSH levels cannot be explained by loss in peritoneal dialysate and may be due to impairment of pituitary function. PMID- 10682116 TI - Response to early measles-mumps-rubella vaccination in infants with chronic renal failure and/or receiving peritoneal dialysis. AB - Achieving immunity to childhood viral infections before renal transplantation is crucial. However, children with chronic renal failure (CRF) may respond poorly to vaccination, making it difficult to achieve immunity before transplantation, particularly if they will require transplantation at a young age. To address this problem, we developed a protocol of early measles-mumps-rubella (MMR) vaccination in infants with CRF. Nine infants received MMR vaccine at a mean age of 11.6 +/- 2.5 months. When vaccinated, 6 of the children (67%) were on peritoneal dialysis, and 3 (33%) had CRF [glomerular filtration rate (GFR) < 30 mL/min/1.73 m2]. Eight patients were later transplanted at a mean age of 16.8 +/- 4.8 months. Titers were measured before transplantation in all patients. Response to vaccination was excellent, with 89% developing immunity to measles, 88% developing immunity to mumps, 100% developing immunity to rubella, and 88% developing immunity to all three components of the vaccine. These response rates were equivalent to, or slightly better than, those previously reported by Schulman for older children (19 +/- 6 months) on dialysis: 80% for measles, 50% for mumps, 100% for rubella, and 30% for all three components. We conclude that early MMR vaccination induces immunity in most infants with CRF, even those on peritoneal dialysis. Response rates are similar to those previously reported in older children. This approach may help to facilitate transplantation in young infants by achieving immunity earlier than traditional vaccination schedules. PMID- 10682117 TI - Effects of recombinant human erythropoietin on physiological inhibitors of coagulation in children on continuous ambulatory peritoneal dialysis. AB - The effects of recombinant human erythropoietin (rHuEPO) on plasma and peritoneal effluent levels of antithrombin III (AT-III), protein C (PC) activity, and protein S (PS) activity were evaluated in 10 uremic children on continuous ambulatory peritoneal dialysis (CAPD). The findings were compared with values obtained from ten healthy children. Levels of AT-III and of PC and PS activity in plasma and peritoneal effluent were measured before, and at 8 and 12 weeks after, rHuEPO treatment. Baseline levels of AT-III and PC activity in plasma were lower than the control values. Levels of PC activity increased during the trial, while levels of AT-III remained unchanged, and levels of PS activity decreased. Baseline levels of PC activity in peritoneal effluent were lower than those obtained during rHuEPO treatment, while no change in peritoneal levels of PS activity and AT-III was observed after rHuEPO treatment. A significant positive correlation was seen between plasma and peritoneal levels of PC activity at baseline. A significant positive correlation was also seen between plasma levels of PS activity and hemoglobin at week 12, and a significant negative correlation between plasma levels of AT-III and albumin at week 8. No correlation was found between the plasma natural coagulation inhibitors and CAPD duration. These results suggest that plasma PS activity can be decreased, and plasma PC activity increased, by rHuEPO treatment in children. PMID- 10682118 TI - Effects of recombinant human erythropoietin on fibrinolytic system in children on continuous ambulatory peritoneal dialysis. AB - We studied tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) levels in plasma and peritoneal effluent in 10 children on continuous ambulatory peritoneal dialysis (CAPD) before, and 8 and 12 weeks after, treatment with recombinant human erythropoietin (rHuEPO). Plasma t-PA and PAI-1 levels were lower in patients than in controls during the study. The plasma t-PA levels were increased by rHuEPO treatment. Although PAI-1 levels showed a tendency to increase in the early phase of rHuEPO treatment, they later returned to near baseline levels. Peritoneal effluent t-PA levels were decreased at week 8 of treatment, but returned to baseline levels at week 12. Peritoneal effluent PAI 1 levels were not changed by the rHuEPO treatment. No correlation was observed between levels of t-PA and PAI-1 in plasma and in peritoneal effluent. No correlation was seen between plasma PAI-1 levels and duration of CAPD. A significant negative correlation was found between the plasma PAI-1 levels and hemoglobin levels at week 8 and week 12. These results suggest that plasma t-PA levels can be increased by rHuEPO treatment, while plasma PAI-1 levels are associated with hemoglobin levels. PMID- 10682119 TI - Middle molecules in peritoneal equilibration test as a marker of peritoneal stress in children on continuous peritoneal dialysis. AB - At 1 month, 3 months, 6 months, and more than 6 months after healed peritonitis, we evaluated repeated peritoneal equilibration tests (PETs) for small molecules such as urea, and middle molecules such as cystatin C, beta 2-microglobulin, and alpha 1-microglobulin. We analyzed a total of 104 PETs in 21 children aged 1.7 18.6 years (median: 9.9 years). Equilibration quotients (D/P)--that is, substrate concentration in dialysis fluid (D) divided by substrate concentration in plasma (P)--were calculated after a dwell time of 4 hours. The D/P for urea did not change after healed peritonitis. In a cross-sectional study, the D/P for middle molecules showed an increase in peritoneal permeability between 3 months and 6 months after a healed peritonitis. In a consecutive follow-up of 4 patients for more than 6 months, beta 2-microglobulin and, more impressively, alpha 1 microglobulin showed a statistically significant increase in D/P (p < 0.05) 3 months after a healed peritonitis. All differences seen were completely reversible after more than 6 months, showing that peritoneal function is rather stable if peritonitis is healed. It is noteworthy that peritoneal dysfunction lasts for up to 6 months after a completely healed peritonitis. This period might be a vulnerable phase in continuation of peritoneal dialysis. PMID- 10682120 TI - Residual peritoneal volume and body size in children on peritoneal dialysis. [Members of the Mid European Pediatric Peritoneal Dialysis Study Group (MEPPS)]. AB - Residual peritoneal volume may play an important role in dialysis efficacy and abdominal compliance in patients on chronic peritoneal dialysis (CPD). In children on CPD, the relationship between residual peritoneal volume and different measures of body size, as well as the day-to-day variability of residual volume, have not been established. We therefore investigated, on two consecutive days, residual peritoneal volume in 25 children on CPD, using the dextran dilution technique. Residual volume was linearly correlated with body size. Residual volume was independent of body size when normalized to body surface area, but decreased with increasing body size when normalized to body weight (r = -0.62, p < 0.001). Mean residual volume was 79 +/- 25 mL/m2, with an intra-individual day-to-day coefficient of variation of 21% +/- 15%. Residual volume was not correlated with the duration of PD, frequency of peritonitis, or peritoneal permeability as estimated by D/P creatinine or D/D0 glucose. In conclusion, residual peritoneal volume is constant across the pediatric age range when normalized to body surface area. It accounts for approximately 8% of the usual fill volume in patients on CPD. Residual volume is not a major confounder of the transport status estimation obtained by peritoneal equilibration test. PMID- 10682121 TI - Can dialysis adequacy be achieved by tailoring the dialysis prescription in an Asian pediatric population on nightly intermittent peritoneal dialysis? AB - This study was undertaken to determine whether tailoring the dialysis prescription in Asian children on nightly intermittent peritoneal dialysis (NIPD), without adding high-dose therapy for cost-savings purposes, was able to achieve dialysis adequacy and improvement in nutrition. Eight children (age range: 5.5-20 years) on NIPD for a mean of 2.1 +/- 0.6 years, were studied at baseline and at 3 months and 9 months after their dialysis dose was tailored. Dialysis adequacy was measured by weekly Kt/Vurea and creatinine clearance (CCr). Fat-free mass (FFM) and percent body fat (%FAT) as measured by bioelectrical impedance, together with anthropometric measurements, serum total protein, and albumin, were used as indicators of nutrition. After the dialysis prescription was tailored, the mean weekly Kt/Vurea increased from 1.89 +/- 0.35 to 2.12 +/- 0.54 at 9 months, and total CCr increased from 36.4 +/- 11.51 L/1.73 m2 to 48.30 +/- 14.30 L/1.73 m2. The increase occurred despite a decline in residual renal function and was attributable to significant improvements in the peritoneal clearances of urea and creatinine (p < 0.05). The mid arm muscle circumference (MAMC) increased significantly (p = 0.006), while FFM increased from 25.68 +/- 7.92 kg to 26.95 +/- 9.83 kg, and %FAT decreased from 21.56% +/- 8.41% to 18.66% +/- 8.16%. The increase in FFM correlated significantly with a decrease in serum creatinine (r = -0.94, p = 0.005). In conclusion, tailoring the dialysis prescription in NIPD, without adding high-dose therapy, resulted in a trend of improvement in dialysis adequacy and some nutritional parameters. PMID- 10682122 TI - Retroviruses et opportunistic infections '99. PMID- 10682123 TI - Analysis of hepatitis B virus quasispecies changes during emergence and reversion of lamivudine resistance in liver transplantation. AB - This report describes nucleotide sequence analysis of part of the polymerase gene of hepatitis B virus (HBV) during the development of lamivudine-resistant HBV in five patients who received lamivudine treatment in conjunction with liver transplantation. Samples from patients were analysed before, during and after drug treatment in conjunction with serum HBV quantification by PCR. Lamivudine resistance was found to be associated with L526M and M550V changes in two patients and M550I change in three patients. Other changes associated with lamivudine resistance in some patients were V509I, A546V, S565A and A568T. The effects on HBV surface antigen are also described. Some patients were subsequently treated with famciclovir and/or ganciclovir with variable outcomes. In two out of three patients who stopped lamivudine treatment, reversion (partial or complete) to wild-type virus was observed after about 5 months. In contrast, a complex mixture of mutant viruses emerged in a third patient who stopped lamivudine treatment. PMID- 10682124 TI - Interferon-alpha therapy exerts selective pressure on hepatitis C virus quasispecies equilibrium. AB - Two patients with chronic hepatitis C virus (HCV) genotype 2b infection were studied. They responded biochemically to interferon (IFN) but had early virological and later biochemical relapse. The HCV quasispecies equilibrium in these patients was studied by a combination of cloning, sequencing and construction of phylogenetic trees. Another patient with chronic HCV genotype 2b infection was followed every 6 months for 30 months (including one episode of biochemical exacerbation) to serve as the control. Quasispecies equilibrium drifted during IFN therapy but moved back in the direction of the original equilibrium during biochemical relapse. In the control patient, there was no significant drifting throughout the follow-up period. These data suggest that IFN therapy exerts selective pressure on HCV quasispecies equilibrium. PMID- 10682125 TI - Stavudine resistance: an update on susceptibility following prolonged therapy. AB - The current report summarizes the available published and unpublished data from several investigators on resistance in clinical isolates following prolonged stavudine therapy. Results suggest that stavudine resistance is both modest in degree and infrequent in appearance. Phenotypic evaluation of 61 patients on stavudine therapy showed only modest changes in drug sensitivity following up to 29 months of treatment. The post-treatment isolates from 15 patients exhibited an increase in EC50 value > fourfold (level above variability of assay) when compared with the corresponding pretreatment isolates. However, the vast majority (11) of these pretreatment isolates either had unexpectedly low EC50 levels and/or had post-treatment isolates that lacked any amino acid changes within their reverse transcriptase (RT) gene to account for the observed change in sensitivity. Of the four remaining isolates, two appeared to have a multi resistant phenotype to several nucleoside analogues and two had no detectable RT amino acid changes to account for the observed change in stavudine sensitivity. To date, clinical HIV-1 isolates displaying stavudine-specific resistance have yet to be reported. Furthermore, full or partial RT sequence analysis of 194 post treatment isolates failed to identify any consistent amino acid changes. The strain-specific V75T mutation reported to confer stavudine resistance to the HXB2 HIV-1 strain in vitro, was found in only six isolates and did not correlate with stavudine resistance. This low incidence of stavudine resistance is in striking contrast to that observed with other nucleoside analogues and further supports the use of stavudine in first-line combination therapy for HIV patients. PMID- 10682126 TI - Induction and maintenance treatment regimens for HIV-1 infection in vitro. AB - Can aggressive anti-human immunodeficiency virus (HIV) induction regimens be simplified after sufficient virus suppression is achieved? In vitro studies were conducted to evaluate the hypothesis that aggressive induction regimens could be followed by less aggressive maintenance regimens. A clinical HIV-1 isolate and lymphoblastoid cell line (H9) were employed. Virus multiplicities were varied, as were drug inhibitory concentrations (IC90, IC99) and induction periods (1, 2 and 3 weeks) of a three-drug regimen (zidovudine plus lamivudine and indinavir), following which maintenance regimens (no drug, zidovudine alone, indinavir alone, zidovudine plus lamivudine) were employed. After 1 week inductions at IC99 concentrations, viral rebound occurred on none or one-drug maintenance regimens but not on a two-drug regimen. After 2 week inductions, viral rebound occurred with no-drug maintenance, but not with one- and two-drug regimens. After 3 week inductions, viral rebound did not occur in zero-, one-, or two-drug maintenance regimens, although HIV-1 DNA persisted in cultured cells. These studies suggest that although some induction-maintenance regimens will fail, after a sufficient period of HIV-1 suppression with a three-drug antiretroviral regimen, maintenance on fewer drugs may be feasible. PMID- 10682127 TI - Quality of life outcomes of saquinavir, zalcitabine and combination saquinavir plus zalcitabine therapy for adults with advanced HIV infection with CD4 counts between 50 and 300 cells/mm3. AB - BACKGROUND: Benefits in patient health-related quality of life (HRQL) have not yet been demonstrated for combination antiretroviral therapy with protease inhibitors and nucleoside analogues. This double-blind study evaluated zalcitabine or saquinavir monotherapy and combination saquinavir plus zalcitabine therapy on HROL of human immunodeficiency virus (HIV)-infected adults. METHODS: 940 HIV-infected patients (CD4 counts 50-300 cells/mm3) who had discontinued zidovudine therapy (for intolerance or treatment failure) were randomized to one of three regimens: zalcitabine 0.75 mg every 8 h; saquinavir 600 mg every 8 h; or combination zalcitabine 0.75 mg plus saquinavir 600 mg every 8 hours. HRQL was measured at baseline, 24 and 48 weeks using the Medical Outcome Study HIV Health Survey (MOS-HIV). The primary endpoints were the physical and mental health summary scores (PHS; MHS) of the MOS-HIV as well as a global visual analogue scale (VAS) score. RESULTS: After 24 weeks, the zalcitabine-treated patients demonstrated significantly greater decreases in PHS scores (-4.4 +/- 0.6; saquinavir: -1.3 +/- 0.6; zalcitabine plus saquinavir: -1.7 +/- 0.6; P < 0.0001) and MHS scores (-2.2 +/- 0.5; saquinavir: -1.0 +/- 0.5; zalcitabine plus saquinavir: -0.5 +/- 0.5; P = 0.032) compared to saquinavir and zalcitabine plus saquinavir treated patients. No differences were observed on the VAS (P = 0.172). Nine of 10 MOS-HIV subscales demonstrated results consistent with the primary endpoints. After 48 weeks, a statistically significant difference between the saquinavir-treated groups and the zalcitabine monotherapy group was observed for PHS scores (zalcitabine: -5.8 +/- 0.6; saquinavir: -4.1 +/- 0.6; zalcitabine plus saquinavir: -3.5 +/- 0.6; P = 0.014). CONCLUSIONS: Saquinavir monotherapy and combination saquinavir plus zalcitabine demonstrated a benefit in HRQL relative to zalcitabine monotherapy in patients with prior zidovudine therapy. The HRQL findings are concordant with improved survival and reduced clinical progression of HIV infection found in this study. PMID- 10682128 TI - Presence of genotypic resistance in nucleoside analogue-treated HIV-1-infected patients with undetectable viral load. AB - Patients harbouring drug-resistance viruses usually suffer a rise in serum viraemia after a variable period of time. We have investigated the relationship between the appearance of resistant genotypes and the viral load of each patient after treatment. Our objective was to assess the association between human immunodeficiency virus (HIV) RNA plasma levels and the number of drug resistance associated point mutations after treatment. A total of 150 patients from three reference centres in Spain (Madrid, Barcelona and Seville) from a previous study (Erase Study) were included. Patients had at that time undergone antiretroviral treatment with nucleoside analogues for at least 1 year (zidovudine/didanosine; zidovudine/zalcitabine; zidovudine/zalcitabine/lamivudine; zidovudine/didanosine/lamivudine). In this study, plasma viraemia in these patients was quantified and a line probe assay was used to determine the genotype of the virus. Viral load was significantly higher in patients harbouring virus with more than three mutations than in those individuals who harboured wild-type strains (P < 0.05). Surprisingly, when patients with viral load < 500 copies/ml (13/150) were analysed, only two carried wild-type strains, whereas three had virus with more than three point mutations. The viral load of six samples was assayed using an ultrasensitive test (detection limit < 20 copies/ml). Of the three samples where viral load was < 20 copies/ml, one patient harboured wild type virus, whereas two carried mutant virus strains. These results suggest that even in patients with undetectable viral loads by conventional methods, viral replication may continue and mutations develop. Therefore, standard values of plasma viraemia for measuring the effectiveness of the treatment should be reconsidered when patients are on antiviral regimens of just two or three nucleoside analogues. PMID- 10682129 TI - Acute hypersensitivity with delavirdine. PMID- 10682130 TI - Finding a role for zalcitabine in the HAART era. AB - Zalcitabine (ddC) is a nucleoside analogue reverse transcriptase inhibitor with demonstrated clinical benefit in combination use. More widespread use of zalcitabine has been limited by a number of factors including peripheral neuropathy and three times daily dosing. However, screening for the risk factors for peripheral neuropathy may enable a reduction in the incidence of neuropathy to below 10%. Additionally, new data on the use of zalcitabine twice daily suggest, based on the long intracellular half-life of the active triphosphate, that this is feasible. Additionally, while limited data exist for zalcitabine in true HAART combinations, data from small trials suggest a similar proportion of responders to standard HAART regimens. PMID- 10682131 TI - Meta-analysis of antiretroviral effects on HIV-1 RNA, CD4 cell count and progression to AIDS or death. AB - There is uncertainty as to how the effects of antiretroviral treatments on human immunodeficiency virus type 1 (HIV-1) RNA levels and CD4 cell counts can predict reductions in clinical progression to AIDS or death. A meta-analysis was conducted for 27 pairwise comparisons of antiretroviral treatments in 15 randomized trials of antiretroviral treatments. For each trial, three measures of treatment effect were used: (i) 16 week change from baseline in HIV-1 RNA; (ii) 16 week change from baseline in CD4 cell count; and (iii) rate of clinical progression. Treatments which caused greater increases in CD4 cell count and greater reductions in HIV-1 RNA were more effective at reducing the rate of clinical progression (P < 0.05 for each comparison). However, there was variability in the consistency of this correlation between different trials and treatments. The results support the use of both CD4 count and HIV-1 RNA levels as the primary markers of the efficacy of antiretroviral treatment. PMID- 10682132 TI - Management of antiretroviral therapy for HIV infection: modelling when to change therapy. AB - OBJECTIVE: To evaluate four strategies for monitoring plasma HIV RNA levels and/or resistance genotypes to decide when to change antiretroviral therapy. The strategies include: (i) 1997 guidelines recommending a therapy switch when plasma RNA exceeds a threshold level; (ii) a viral load policy, using a fixed increase in viral load as the trigger; (iii) a genotype policy, requiring a smaller viral rebound than (ii) and detection of genotypic resistance before switching; and (iv) a proactive policy, switching drug regimens at a predetermined time if viral load has not rebounded. DESIGN AND SETTING: A Monte Carlo simulation tracks patients' viral loads and presence of opportunistic infection during therapy. The model uses clinical and virological data and statistical variation in patient parameters for the evaluation of therapeutic strategies. MAIN OUTCOME MEASURES: To determine which strategies minimize viral rebound detection delay while maintaining a low (prespecified) probability of switching therapy before rebound. RESULTS: 1997 Guidelines and the viral load policy create lengthy delays in detection of rebound, particularly when patients are drug-naive and the detection limit of the viral load assay is 500 copies/ml. A detection limit of 20 copies/ml decreases this delay substantially. Genotyping achieves only minor additional delay reductions. Of the strategies tested, the proactive policy leads to the shortest delays. CONCLUSIONS: This model indicates that prolonged periods may be required for viral load to rebound to detectable levels following prolonged suppression. Proactive switching produces the best outcome in our model because it may reduce the duration of viral replication under pressure of a failing regimen before detection of viral rebound. This strategy should be evaluated in clinical trials. PMID- 10682133 TI - Improvement in immune function due to treatment with indinavir despite severe immune deficiency. AB - To study the virological, immunological and clinical effects of the protease inhibitor indinavir in human immunodeficiency virus (HIV)-infected patients with CD4 counts < 50 cells/mm3, indinavir was added to prior treatment with nucleoside analogues in a prospective open-label study in 23 HIV-infected patients with median CD4 count of 10 cells/mm3 and median serum HIV-1 RNA load of 27,508 copies/ml. Addition of indinavir induced a decrease in HIV-1 RNA levels to < 400 copies/ml in 15 patients that was maintained until week 36 of the study in 8 (35%) patients. The median increase in CD4 cell counts was 92 cells/mm3 (range 55 258 cells/mm3) and in CD8 counts was 245 cells/mm3 (range 51-1552 cells/mm3) at week 30. The treatment induced a significant CD8 T cell expansion, consisting in the first 6 weeks of predominantly memory CD45RO+ cells and followed by expansion of naive cells from week 12 on, and a significant decrease in the proportion of activated CD8/CD38 cells. In addition, significant increases in T cell proliferation following stimulation with phytohaemagglutinin and significant decreases in the rates of spontaneous apoptosis of CD4+ and CD8+ T cells were observed. In conclusion, the addition of indinavir induced restoration of both memory and naive CD8 T cells. Corresponding evidence of improving T cell function, as assessed by enhanced lymphoproliferative capacity and diminished propensity to undergo apoptosis, provides evidence for treatment-induced regeneration of immune function even in patients with severe immunodeficiency. PMID- 10682134 TI - Prediction of HIV-1 RNA suppression and its durability during treatment with zidovudine/lamivudine. AB - To predict the probability of long-term viral suppression during treatment with zidovudine and lamivudine, human immunodeficiency virus type 1 (HIV-1) RNA values were retrospectively pooled for 1083 patients from six randomized, double-blind clinical trials. All analyses of HIV-1 RNA were obtained using the Roche Amplicor assay or its earlier prototype. Time to loss of response was evaluated by Kaplan Meier analysis; Cox proportional hazards models were used to assess the influence of baseline variables. Among 523 patients with < or = 6 months of prior zidovudine treatment, the probability of HIV-1 RNA suppression below 400 copies/ml at 48 weeks was 71% in those with baseline HIV-1 RNA < 5000 copies/ml, but only 14% in those with HIV-1 RNA between 50,000 and 200,000 copies/ml. Among 560 patients with > 6 months of prior zidovudine treatment, the rates of sustained viral suppression were lower, but also significantly associated with the baseline HIV-1 RNA. Multivariate analyses showed no independent effect of CD4 cell count, age, sex, race, or CDC disease stage on the probability of sustained HIV-1 RNA suppression. When patients with < or = 6 months of prior therapy were stratified based on the magnitude of HIV-1 RNA nadir achieved during treatment, those who reached a nadir of < 400 copies/ml retained this response for significantly longer time periods than the ones who only achieved partial viral suppression. In conclusion, baseline HIV-1 RNA levels and the duration of prior zidovudine therapy strongly predict the antiretroviral efficacy of zidovudine/lamivudine. The baseline parameters should influence the choice of the antiretroviral regimen. PMID- 10682135 TI - Diminished HIV-1 sensitivity to stavudine in patients on prolonged therapy occurs only at low levels and cannot be attributed to any single amino acid substitution in reverse transcriptase. AB - To study the extent to which phenotypic resistance to stavudine occurs under therapy, we studied 18 pairs of human immunodeficiency virus type 1 (HIV-1) isolates from patients both prior to and following 24-48 weeks of treatment with stavudine monotherapy or stavudine in combination with either didanosine or lamivudine. We also used a nested polymerase chain reaction (PCR) assay to probe for the presence of specific mutations associated in culture with stavudine resistance. The results showed that resistance to stavudine (approximately 3-10 fold) was observed in nine of ten cases of monotherapy, in three of four cases of therapy involving both stavudine and didanosine, and in two of four cases involving stavudine and lamivudine. Viruses from the four patients receiving stavudine plus didanosine became resistant to didanosine in only one instance while the use of lamivudine plus stavudine yielded resistance to lamivudine each time. Whereas changes in the reverse transcriptase (RT) genes of resistant isolates were frequently observed, two mutations, previously identified with stavudine resistance in tissue culture (i.e., V75T and I50T), could not be identified in the clinical samples by either direct sequencing of the RT gene or by PCR amplification. Thus, resistance to stavudine can occur, albeit at low levels, in the context of prolonged therapy with this drug but is not associated with specific mutations in HIV RT at either codons 75 or 50 in clinical samples. PMID- 10682136 TI - Genotypic and phenotypic resistance to stavudine after long-term monotherapy. BMS 020 Spanish Study Group. AB - Protocol BMS 020 was a double-blind, prospective clinical trial in which two different doses of stavudine (20 and 40 mg twice daily) were compared in human immunodeficiency virus (HIV)-infected patients with previous exposure to zidovudine for longer than 16 weeks. Genotypic and phenotypic resistance to both zidovudine and stavudine were examined after at least 2 years of stavudine monotherapy. None of 35 tested individuals harboured the codon 50 and/or 75 mutations previously described to be associated with stavudine resistance. However, more than 80% of the individuals carried mutations associated with zidovudine resistance, despite all these patients having stopped zidovudine at least 2 years earlier. Significant phenotypic resistance to stavudine was observed only in 2 of 5 tested individuals, although IC50 values were increased only 6.6- and 9.2-fold, respectively. These two patients had suffered a decline in their CD4 count, and one of them had high levels of plasma viraemia. The sequence analysis of the reverse transcriptase (RT) gene (aa 30 to 240) in these five patients revealed no changes that could be involved in stavudine resistance. In contrast, and despite having stopped treatment with zidovudine more than 2 years before, phenotypic resistance to zidovudine was observed in all five subjects, with IC50 values raised by more than 75-fold in all of them. Moreover, all harboured codon substitutions within the RT gene associated with zidovudine resistance, and these mutations remained in viral genomes examined after virus co culture, suggesting that they provided some biological advantage to mutants, even in the absence of drug pressure. In conclusion, both genotypic and phenotypic resistance to stavudine seem to be a rare event in patients exposed to the drug, even after long periods of exposure. PMID- 10682137 TI - Quantitative p24 antigenaemia for monitoring response to antiretroviral therapy in HIV-1 group O-infected patients. AB - Failure to recognize infection caused by human immunodeficiency virus type 1 (HIV 1) group O variants has been described using both serological and genetic techniques. Moreover, the monitoring of response to antiretroviral therapy is difficult in persons carrying this infection since most currently available tests for quantifying viral load are not reliable for group O viruses. Considering the low level of divergence between the p24 proteins of group M and O viruses, we have examined whether the quantification of circulating p24 antigenaemia might be used as a surrogate marker of response to therapy in three subjects with HIV-1 group O infection treated with antiretroviral drugs. In summary, all three patients showed a significant decline in circulating plasma p24 antigenaemia, although only one achieved undetectable levels. The decline in p24 antigenaemia was parallel to an increase in the CD4 count and was associated with an improvement in clinical status. PMID- 10682138 TI - Anti-HIV antiviral activity of stavudine in a thymidine kinase-deficient cellular line. AB - Stavudine (d4T) is a potent inhibitor of human immunodeficiency virus type 1 (HIV 1) reverse transcriptase. It is known that stavudine is metabolized in cells to the mono-, di- and triphosphate nucleotides but the enzymes responsible for its phosphorylation are as yet unidentified. In particular, there are conflicting results concerning the role of thymidine kinase 1 (TK1) in stavudine metabolism. To gain new insights into this phenomenon we analysed the antiviral activity of stavudine in a TK1-deficient, resistant cell line. The results indicate that TK1 is responsible for the phosphorylation of stavudine but it is not the only enzyme involved in its activation. The other enzyme(s) that might be involved in the metabolism of stavudine, however, are not able to phosphorylate stavudine with the same efficiency as TK1. Since it has been shown that prolonged treatment with zidovudine may induce an in vivo defect in TK1 activity, it is tempting to speculate that patients treated for a long time with zidovudine could be resistant to further treatment with stavudine. PMID- 10682139 TI - Loss of lamivudine resistance in a zidovudine and lamivudine dual-resistant HIV-1 isolate after discontinuation of in vitro lamivudine drug pressure. AB - We examined the in vitro phenotypic and genotypic profiles of an extensively passaged human immunodeficiency virus type 1 clinical isolate which has been selected for lamivudine resistance, with an M184V mutation in a zidovudine resistant genetic background, and then cultured with zidovudine alone. Our passaging strategy led to a decrease in lamivudine IC50 values, which were comparable to those prior to lamivudine exposure, and the genotypic restoration of the wild-type sequence at codon 184 of reverse transcriptase. PMID- 10682140 TI - Kinetics of productive and latent HIV infection in lymphatic tissue and peripheral blood during triple-drug combination therapy with or without additional interleukin-2. AB - OBJECTIVE: To study decay rates of productively and latently infected cells in peripheral blood and lymph nodes during triple antiretroviral therapy and the possible impact of interleukin-2 (IL-2) on viral kinetics. METHODS: In this non randomized study, nine antiretroviral-naive HIV-positive patients received either saquinavir hard gel capsules 2400 mg three times daily (group I; four patients) or saquinavir soft gel capsules 1200 mg three times daily and IL-2 (group II), in both cases together with two nucleoside analogues. Plasma viraemia and lymphocyte subsets were analysed. Axillary lymph nodes were excised before and after 12 weeks of therapy. Lymph node sections were examined by in situ hybridization for HIV RNA, and productively infected cells were counted. Infection rates of FACS sorted CD3, CD4 lymph node and peripheral blood mononuclear cells were determined by nested DNA PCR. RESULTS: Baseline plasma HIV RNA levels ranged from < 25 to > 1 x 10(6) copies/ml and remained undetectable throughout the study in one patient in group I. Plasma viraemia became undetectable after 3 months in four patients (three in group I). Productively infected cells were markedly reduced in the follow-up lymph node specimens. HIV DNA-positive CD4 T cells were reduced in lymphoid tissue and peripheral blood in all six evaluable patients. There were no significant differences between the groups in the clearance rates of plasma virus and of HIV DNA-positive cells. CONCLUSIONS: Combined antiretroviral therapy rapidly suppressed active HIV replication in plasma and lymphoid tissue. Latently infected cells were cleared at a slower rate. Viral clearance did not appear to be markedly affected by additional IL-2 therapy. PMID- 10682141 TI - Low plasma concentrations of indinavir are related to virological treatment failure in HIV-1-infected patients on indinavir-containing triple therapy. AB - All human immunodeficiency virus type 1 (HIV-1)-infected patients who started to use indinavir (800 mg three times a day) as part of their triple drug regimen were included in a study to determine the importance of low plasma concentrations of indinavir as a cause of virological treatment failure. The indinavir concentration and a number of patient characteristics at baseline were tested as risk factors for virological treatment failure (defined as a viral load above 200 copies/ml after 24 weeks of treatment) in univariate and multivariate analyses; 65 patients were included. Virological treatment failure occurred in 36.9% of the patients. Multivariate analysis showed that a low plasma concentration of indinavir (odds ratio 0.1), a high viral load at baseline (odds ratio 2.6) and pretreatment with another protease inhibitor (odds ratio 10.0) were independent factors related to virological treatment failure. Monitoring of indinavir plasma concentrations may be an important tool for the optimization of triple drug combination therapy. PMID- 10682142 TI - Evaluation of mixtures of wild-type HIV-1 and HIV-1 with resistance point mutations against reverse transcriptase inhibitors. AB - The presence of resistance-related mutations in 185 serial proviral DNA samples from 108 HIV-infected patients was monitored using the line probe assay (LiPA). The proportions of wild-type and mutant virus in each sample were determined. Subsequent samples from the same patient were analysed. Resistance mutations were detected in 58 of 108 patients studied (53.7%), 53 of 73 (72.6%) treated with antivirals and 5 of 35 (14.2%) untreated. The mutations were against zidovudine (51), lamivudine (1), zidovudine and lamivudine (4), zidovudine and zalcitabine (1) and zidovudine and didanosine (1). Among the 58 patients with resistant virus, 168 related mutations were observed: 161 to zidovudine (90 in codon 70, 25 in codon 41 and 46 in codon 215), 5 to lamivudine (codon 184), 1 to zalcitabine (codon 69) and 1 to didanosine (codon 74). Mixtures of wild-type and resistant mutants were detected in 76 of 90 (84.4%) mutated at codon 70, 28 of 46 (60.8%) mutated at codon 215 and in 21 of 25 (84%) mutated at codon 41. The mutations at codon 184 were mixtures of wild-type and resistant in 4 of 5 samples. The agreement between LiPA and sequencing was 96.5%. LiPA was more sensitive for the detection of mutants that were present at low frequency. The analysis of sequential samples from the same patient allowed evaluation of the dynamics of appearance of the resistant mutants. PMID- 10682143 TI - Opportunistic infections shortly after beginning highly active antiretroviral therapy. AB - The clinical benefit of highly active antiretroviral therapy (HAART) has been attributed to its suppression of viral replication and improvement in the CD4 lymphocyte count. However, the development of clinical symptoms secondary to previously silent opportunistic pathogens shortly after beginning HAART has been reported as a distinct clinical syndrome and seems to be associated with inflammatory phenomena surrounding a rapid restoration of the immune system in previously immunosuppressed patients. Herein, we report nine (3.6%) episodes of opportunistic infections (OI) in 247 human immunodeficiency virus (HIV)-infected patients undergoing HAART in a reference HIV/AIDS institution located in Madrid, Spain. In all instances, OI clustered within the first 3 months after beginning HAART. Episodes of cerebral toxoplasmosis (three cases), Pneumocystis carinii pneumonia (two cases), and herpes zoster (two cases) occurred in persons without a previous AIDS-defining illness, in addition a relapse of cytomegalovirus retinitis and a rebound in Kaposi's sarcoma were seen, respectively, in another two patients. Four of the nine subjects had a CD4 count above 200 cells/mm3 before HAART began. Of these, one developed Pneumocystis pneumonia and one other cerebral toxoplasmosis. In conclusion, prophylaxis and close clinical monitoring of HIV-infected patients should be considered for the first 3 months after beginning HAART, even for subjects without severe immunosuppression. PMID- 10682144 TI - Combination treatment for hepatitis C can be hazardous in patients co-infected with HIV. PMID- 10682145 TI - Antiviral drug resistance: from the laboratory to the patient. PMID- 10682146 TI - Cytomegalovirus drug resistance. AB - Clinical resistance of cytomegalovirus (CMV) against the currently licensed antiviral drugs is becoming an increasingly recognized problem. This review focuses on the molecular basis of resistance and describes mutations in the UL54 DNA polymerase leading to resistance against cidofovir, foscarnet and ganciclovir. The review highlights two important developments in our appreciation of resistance. Firstly, the use of more rapid molecular based assays to detect genotypic resistance and secondly, the relationship between resistance profiles in multiple organ systems of the same host. Finally, the changing face of CMV disease in the era of highly active antiviral chemotherapy is considered with respect to its impact on the frequency of CMV resistance in the clinic. PMID- 10682147 TI - Hepatitis B virus antiviral drug resistance: from the laboratory to the patient. AB - The development and application of nucleoside (and nucleotide) analogues for the treatment of chronic hepatitis B infection will transform the management of this condition. For instance, treatment with lamivudine effects a dramatic and measurable reduction of serum virus titre. This is associated with biochemical and histological improvements. Unfortunately, for the majority, replication resumes when treatment is withdrawn. Prolonged lamividine treatment may be associated with the emergence of drug-resistant species with specific polymerase mutations. Compared with the observed rate for the development of drug resistance observed during monotherapy of HIV infection, resistance is slow to emerge during treatment of hepatitis B. The rate of emergence might be dependent on the rate of infected hepatocyte turnover, which is extremely variable in chronic HBV infection (and significantly slower than infected lymphocyte turnover during HIV infection). Preliminary data suggest that pretreatment serum virus titre may be an important predictor of the development of drug resistance, an observation consistent with preexistance of the resistant virus in the hepatitis B virus quasispecies. Akin to developments in antiviral treatment of HIV infection, further progress in the treatment of chronic hepatitis B will depend on the development of drugs for use in combination therapy. PMID- 10682148 TI - Laboratory markers of antiviral activity. AB - Quantitative assays for viral nucleic acids have been instrumental in monitoring the response of patients to various antiviral therapies. The level of viraemia is predictive of clinical outcome in that a reduced risk of progression to AIDS or death was observed with lower plasma human immunodeficiency virus (HIV) RNA levels. Rebound in viral levels often signals therapeutic failures, some of which are associated with the development of drug resistance. Quantitative plasma assays for HIV, hepatitis C virus (HCV), cytomegalovirus (CMV) and hepatitis B virus (HBV) have been developed. Over time, modifications to these assays have been required to meet new demands. For example, as antiviral therapies have become more effective, HIV and HCV assays of greater sensitivity are required in order to follow patients for longer periods of time and to fully assess the extent of viral suppression. For HIV-1, a large percentage of patients treated with combination therapies had viral loads that were below the detection limit of the ultrasensitive assay (50 copies/ml). To assess the residual viral burden in this patient population an assay to quantify HIV-1 proviral DNA in peripheral blood mononuclear cells was developed. Studies to date indicate that proviral DNA remains easily detectable despite undetectable plasma RNA and may be useful in monitoring this patient population. To increase assay throughput, a new generation of quantitative assays that will provide real-time detection and a 6 log10 detection range from a single amplification is under development. PMID- 10682149 TI - Effective salvage therapy for HIV-1 infection--an unmet challenge. PMID- 10682150 TI - Safety and efficacy of the neuraminidase inhibitor zanamivir in treating influenza virus infection in adults: results from Japan. GG167 Group. AB - The study was carried out to evaluate the therapeutic effects of zanamivir, a highly selective, potent and specific inhibitor of influenza A and B virus neuraminidases, in adult patients with acute influenza-like illness. Patients who presented within 36 h of the onset of influenza-like symptoms were randomly assigned to receive one of three treatments, twice daily, for 5 days: 10 mg zanamivir powder for inhalation (zanamivir inhalation group), 10 mg zanamivir powder for inhalation plus 6.4 mg zanamivir nasal spray (zanamivir inhalation plus intranasal group) or placebo (placebo group). The primary end point was the time to alleviation of the three major symptoms (fever, headache and myalgia). The secondary end point was the time to alleviation of five influenza symptoms (fever, headache, myalgia, cough and sore throat). One hundred and sixteen patients with influenza-like illness were recruited to the study. No differences were observed between the two groups of patients who received zanamivir (inhalation group or inhalation plus intranasal group). Patients who received zanamivir recovered significantly faster (median 3 days to recovery) than the patients in the placebo group (median 4 days to recovery; P < 0.01). Topically administered zanamivir was well tolerated. This study confirms that in adults, topically administered zanamivir is well tolerated and is effective in reducing the time to alleviation of influenza symptoms. PMID- 10682151 TI - Zidovudine resensitization and dual HIV-1 resistance to zidovudine and lamivudine in the delta lamivudine roll-over study. AB - OBJECTIVE: To study zidovudine resensitization and dual resistance to zidovudine/lamivudine in HIV-1 isolates from nucleoside reverse transcriptase (RT) inhibitor-experienced patients during selective pressure exerted by zidovudine/lamivudine combination therapy. DESIGN AND METHODS: HIV-1 isolates from 29 patients receiving zidovudine/lamivudine combination therapy in the Delta roll-over study were analysed at entry and during a 1 year follow-up period for phenotypic susceptibility to zidovudine and lamivudine in the ANRS PBMC assay. The RT gene from codon 20 to 230 and at codon 333 was analysed by nucleotide sequencing of the corresponding isolates. RESULTS: HIV-1 isolates from 23 of the 29 patients were phenotypically resistant to zidovudine at baseline; 61% of these patients showed significant zidovudine resensitization during follow-up. The zidovudine IC50 value correlated positively with log10 plasma HIV-1 RNA (P = 0.02) and negatively with the CD4 cell count (P = 0.004). Zidovudine resensitization (related to acquisition of the M184V mutation) was transient, with evolution towards dual resistance to zidovudine and lamivudine in 20 of the 29 patients. The phenotype of certain dually resistant isolates coincided with the emergence of multiple mutations in the 5' part of the RT gene. CONCLUSIONS: M184V-mediated zidovudine resensitization of HIV-1 is transient in most patients who are given zidovudine/lamivudine combination therapy when zidovudine resistance has already emerged. The subsequent evolution towards dual phenotypic resistance to zidovudine/lamivudine corresponds to complex genotypic profiles. PMID- 10682152 TI - AVANTI 1: randomized, double-blind trial to evaluate the efficacy and safety of zidovudine plus lamivudine versus zidovudine plus lamivudine plus loviride in HIV infected antiretroviral-naive patients. AVANTI Study Group. AB - The objective of this randomized double-blind, placebo-controlled trial was to investigate the effect of combination antiretroviral therapy on plasma HIV-1 RNA as measured by HIV RNA PCR and to assess the safety and tolerability of such regimens. The trial was carried out in seven European countries, Australia and Canada and involved antiretroviral-naive patients (n = 106) with CD4 counts between 150-300 cells/mm3 (CDC group A) and 150-500 cells/mm3 (CDC group B/C). Patients were randomly assigned to zidovudine (200 mg three times daily) plus lamivudine (300 mg twice daily) or to zidovudine plus lamivudine plus loviride (100 mg three times daily) for 52 weeks. The main outcome measures were degree and duration of reduction of plasma HIV-1 RNA as measured by RNA PCR and the development of drug-related toxicities sufficiently severe to warrant dose modification, interruption or permanent discontinuation. A mild, though statistically significant difference in favour of zidovudine plus lamivudine plus loviride for log10 plasma HIV-1 RNA (P = 0.022), as compared to zidovudine plus lamivudine, was observed using area-under-the-curve minus baseline (AUCMB). An increase in CD4 cell count in the zidovudine plus lamivudine plus loviride group was observed with a median improvement of 124 cells/mm3 at week 52 compared with 70 cells/mm3 in the zidovudine plus lamivudine group (P = 0.06). Both treatment regimens were well tolerated. PMID- 10682153 TI - In vitro selection and characterization of HIV-1 with reduced susceptibility to PMPA. AB - 9-(2-phosphonomethoxypropyl)adenine (PMPA) has demonstrated remarkable anti simian immunodeficiency virus (SIV) activity in macaque models of SIV infection and transmission prevention. Recently, PMPA and its oral prodrug, bis-POC PMPA, have also shown potent anti-human immunodeficiency virus type 1 (HIV-1) activity in Phase I clinical studies. In vitro experiments were performed to address the resistance properties of PMPA. After eight passages in increasing concentrations of PMPA, HIV-1IIIB was able to grow in the presence of 2 microM PMPA, fivefold above the IC50 of PMPA for wild-type parental virus. Sequence analysis of the reverse transcriptase (RT) genes from four of 15 RT clones demonstrated the presence of a K65R substitution in RT and recombinant HIV expressing the K65R RT mutation showed a threefold to fourfold increase in IC50 value for PMPA as compared to wild-type. Additional experiments demonstrated that viruses expressing other nucleoside-associated RT resistance mutations all showed wild type or < threefold reduced susceptibility to PMPA in vitro. Interestingly, lamivudine-resistant viruses expressing the M184V RT mutation showed wild-type to slightly increased susceptibility to PMPA in vitro and addition of the M184V mutation to HIV with the K65R mutation resulted in reversion to wild-type susceptibility for PMPA. In agreement with the cell culture findings, Escherichia coli-expressed K65R RT showed fivefold reduced susceptibility to PMPA diphosphate, the active moiety of PMPA. Furthermore, in combination experiments, PMPA with hydroxyurea showed synergistic inhibition of HIV replication in vitro. The potent antiretroviral activity and favourable resistance profile of PMPA and bis-POC PMPA are being further investigated in ongoing clinical trials. PMID- 10682154 TI - A combination of nucleoside analogues and a protease inhibitor reduces HIV-1 RNA levels in semen: implications for sexual transmission of HIV infection. AB - Direct contact with semen is the major route of sexual acquisition of human immunodeficiency virus (HIV) in homosexual and heterosexual partners of seropositive men. In this study, we show that concentrations of HIV-1 RNA molecules in plasma and semen of seropositive patients are related to the duration and type of antiretroviral agents used in treatment. In patients treated with zidovudine alone, 1, 3 and 6 months after the start of therapy, the mean HIV 1 load in plasma was reduced by 0.57, 0.38 and 0.21 log10 and in semen by 0.66, 0.50 and 0.15 log10, respectively. In patients treated with zidovudine plus didanosine at months 1, 3 and 6, the mean decrease in plasma HIV-1 RNA was 1.40, 1.25 and 1.12 log10 and in semen 1.10, 1.41 and 1.32 log10, respectively. In patients treated with a combination of a protease inhibitor and two nucleoside analogues the mean log10 decrease was 1.77, 1.83, 1.71 and 2.38 log10 in plasma and 1.17, 1.74, 2.19 and 3.02 log10 in semen at 1, 2, 3 and 4 months, respectively. Treatment with a combination of a protease inhibitor and two nucleoside analogues caused a dramatic decrease in cell-free HIV-1 RNA in semen, which is a reliable measure of viral load. These findings could have implications for the sexual transmission of HIV-1. PMID- 10682155 TI - Effect of weekly adefovir (PMEA) infusions on HIV-1 virus load: results of a phase I/II study. AB - The compound 9-(2-phosphonylmethoxyethyl)adenine (adefovir; PMEA) is a potent inhibitor of a number of viruses in vitro, such as human immunodeficiency virus (HIV) type 1 and 2, herpes simplex virus (HSV) type 1 and 2, human papillomavirus virus (HBV) and Epstein-Barr virus (EBV). Adefovir also proved to be effective in vivo against feline immunodeficiency virus (FIV) in cats and simian immunodeficiency virus (SIV) in rhesus monkeys. In an open, non-placebo controlled trial the antiviral activity of weekly doses of adefovir in nine patients with AIDS or AIDS-related complex was studied for a period of 11 weeks. CD4 cell counts at baseline were between 10 and 450 cells/mm3, HIV-1 RNA levels at baseline were between 24,210 copies/ml and 406,197 copies/ml. The drug was administered intravenously at a dose of 1000 mg every week and plasma viral load was assessed at multiple points during the study. Administration of adefovir was tolerated well and no severe side effects were seen. The response to adefovir treatment differed widely between patients. The increase in CD4 cell count at end point ranged from -40 to 120 cell/mm3. The lowest HIV RNA levels were measured after 3-5 days, showing an increase thereafter. The nadir in viral load was achieved after 2 weeks, with a mean viral load decline of 0.7 from baseline. The decrease of the HIV RNA level at end point ranged from -0.3 log10 to 1.8 log10 with a mean decrease of 0.4 log10. Our results indicate that adefovir given intravenously once weekly has a short-lasting initial antiviral effect. The effect of more frequent dosing requires further evaluation. If adefovir is to be useful clinically, it needs to be combined with other antiviral agents. PMID- 10682156 TI - Kinetics of beta-chemokine levels during anti-HIV therapy. AB - Chemokines are pro-inflammatory cytokines that inhibit human immunodeficiency virus type 1 (HIV-1) replication in vitro. We studied the kinetics of the beta chemokines, macrophage inhibitory protein (MIP)-1 alpha, MIP-1 beta, RANTES, and monocyte chemotactic protein (MCP)-1 in plasma during 12 months of antiretroviral therapy in 26 HIV-1-infected patients and in 11 untreated subjects. Eleven patients with moderate immunodeficiency had HIV-1 RNA levels < 50 copies/ml after 1 year, whereas 12 out of 15 patients with severe immunodeficiency had detectable virus. At baseline, MCP-1 levels correlated positively with HIV-1 RNA and DNA levels and inversely with CD4 cell counts. A reverse pattern was seen for the MIP 1 beta levels. No correlation was seen between MIP-1 alpha or RANTES and any of the parameters. Also, there was a dichotomy between the different beta-chemokines in response to therapy. Decreases of MCP-1 and RANTES levels were found, but no durable changes of MIP-1 alpha and MIP-1 beta. The MCP-1 levels rebounded back to baseline after 1 year in the patients who responded virologically, which could possibly reflect an increased immune activation. The biological consequences of the changes in beta-chemokines levels during antiretroviral treatment are still unknown and deserve further studies. PMID- 10682157 TI - Patients failing saquinavir therapy require an early change to indinavir before HIV-1 viral load reaches high levels. AB - Sequential use of antiretroviral therapy with protease inhibitors (PI) is frequently prescribed owing to failure or intolerance of the first selected agent. Controversial data exist about the virological and immunological outcome of patients in whom a change to a second PI regimen is needed. A prospective study of 113 HIV-positive patients (male, 84%; mean age 36 years; previous AIDS defining event, 35%; previous antiretroviral therapy with nucleoside analogues, 94%) who started a saquinavir-containing regimen between March 1996 and March 1997 and had to change to indinavir (n = 79) owing to intolerance, failure or medical criteria was performed. At the time of the switch, median CD4 cell count was 221 cells/mm3 and the HIV RNA level was 3.98 log10 copies/ml. The rate of viral suppression (HIV RNA levels below 200 copies/ml) was 40% at 3 months and 28% at month 6 after indinavir was instituted. In a logistic regression analysis, only the baseline viral load [relative risk (RR), 2.85; 95% confidence interval (CI), 1.31-6.05; P = 0.007] was statistically associated with the lack of viral suppression on indinavir. Although there are not sufficient data about the best therapeutic option if a change in PI-containing regimens therapy is considered, this study supports the recommendation of an early change of the PI-containing regimens, before the HIV-1 viral load reaches high levels. PMID- 10682158 TI - Efficiency of drug resistance genotypic tests in specimens with low HIV viral load. AB - The early recognition of resistance to antiretroviral agents could allow a rapid switch in therapy and therefore avoid the accumulation of mutations and reduce the risk of cross-resistance. However, the efficiency of genotypic tests in specimens with low viral load (VL) is severely compromised since human immunodeficiency virus (HIV) RNA in these samples often goes unrecognized. The frequency of results provided by a line probe assay (LiPA, Murex), a commercially available drug resistance test and a home-made point mutation assay (PMA) for recognizing the codon 151 multidrug-resistance mutation was examined in 664 plasma samples stratified with respect to VL values. Overall, 421 (63%) samples could be interpreted by both LiPA and PMA. The sensitivity decreased as plasma VL lowered: 89% for samples with VL > 10,000 HIV RNA copies/ml, 77% for those with VL between 500 and 10,000 HIV RNA copies/ml and 37% for specimens with VL < 500 HIV RNA copies/ml. A good agreement existed comparing the sensitivity of the home made PMA and LiPA. Although the former tends to produce more results, the difference did not achieve statistical significance. Our results support that new, more sensitive, HIV RNA extraction methods need to be implemented for the rapid recognition of drug-resistant mutants in patients experiencing an early rebound in plasma viraemia. PMID- 10682159 TI - Different outcome in the first two patients with an HIV-1 multinucleoside drug resistant T69SSS insertion in Spain. AB - A novel multidrug-resistance mechanism has been described in human immunodeficiency virus type 1 (HIV-1), which involves the insertion of 6 bp between codons 69 and 70 in the reverse transcriptase (RT) gene. Herein, we report the first two patients in Spain carrying viral populations with the 69-SS insert coupled to the T69S mutation. Both patients were selected because of the lack of signal at positions 69/70 in the LiPA RT test despite being reactive to the remaining probes on the LiPA strip. The presence of the T69SSS complex was confirmed by sequence analysis. A common feature for both subjects was their past history with zidovudine monotherapy and zidovudine plus either didanosine or zalcitabine later on in the presence of persistent virus replication. Remarkably, the introduction of triple therapy in patient 1 soon after the emergence of the insert-containing viral strain produced its total displacement, which correlated with a sustained suppression in viral load. PMID- 10682160 TI - Atherogenesis and its relationship to coronary risk factors. AB - A new model for the development of atherosclerosis is emerging (1,2). This development process, called atherogenesis, is now thought to begin with metabolic dysfunction of the endothelial cells that line the innermost portion of the arterial wall. Endothelial dysfunction precedes visible changes in endothelial structure. Dysfunctional endothelium loses its ability to maintain vascular smooth muscle relaxation and instead promotes vasospasm, chemotaxis and inflammation, platelet aggregation, and diminished clot lysis. Endothelial dysfunction appears to occur diffusely, rather than discretely, in affected vessels. Accordingly, local anatomical interventions, such as bypass surgery or angioplasty, can be expected to have only limited success in the treatment of patients with atherosclerotic disease. More definitive treatments must be directed at the risk factors initiating or enhancing atherogenesis. Such interventions are more likely to be medical than surgical or mechanical. With appropriate understanding of the underlying process of atherogenesis and its clinical manifestations, such medical interventions can be carried out within the boundaries of everyday practice. PMID- 10682161 TI - Clinical diagnosis of lipid disorders. AB - Atherosclerotic cardiovascular disease is a major health problem in the United States. In particular, coronary heart disease (CHD) is the leading cause of death in men and women in the United States, as well as in other industrialized countries. Extensive observational epidemiologic data within and between populations have strongly linked such various factors as untreated hypertension, diabetes, cigarette smoking, and lipid abnormalities to the development of CHD. With respect to lipoprotein parameters, elevated total and low-density lipoprotein cholesterol (LDL-C) and low levels of high-density lipoprotein cholesterol (HDL-C) have been strongly associated with CHD risk. Emerging evidence suggests that other lipoprotein abnormalities also are associated with premature CHD, including elevated levels of lipoprotein(a), triglyceride-rich lipoproteins such as small very-low-density lipoproteins and intermediate-density lipoproteins, small and dense LDL particles, and the magnitude of postprandial lipemia. Extensive primary and secondary clinical trial evidence has established that favorably altering dyslipidemias through diet and a variety of pharmacologic agents produces clear improvements in CHD end points. The extent of this benefit depends on the presence or absence of clinical atherosclerotic disease, as well as other CHD risk factors, and the severity of one or more lipoprotein abnormalities. CHD patients and individuals with multiple risk factors, but free of clinical CHD, derive the greatest absolute benefit from lipid treatment directed at reducing LDL-C. The dyslipidemias that impart high risk are severely elevated LDL-C (> 200 mg/dL), combined high LDL-C and low HDL-C (< 35 mg/dL), and combined hyperlipidemias (non-HDL-C > 200 mg/dL with low HDL). The purpose of this review is to aid the primary care physician in identifying these important dyslipidemias and to critically analyze the relative importance of various lipoproteins on atherosclerotic risk. PMID- 10682162 TI - Primary prevention of coronary disease. AB - In 1995, the latest year for which statistics are available, heart disease, cancer, and stroke continued to be the three leading causes of death in the United States. Notably, however, a wealth of experience has confirmed that hygienic interventions such as diet, exercise, weight loss, and smoking cessation can reduce the toll from heart disease while also reducing the morbidity and mortality associated with stroke and cancer. This chapter will describe the rationale for the primary prevention of coronary heart disease (CHD), review the basic concepts involved in cholesterol screening, and update the reader regarding key preventive measures, such as diet, exercise, and smoking cessation. Also highlighted will be recent clinical trial results suggesting the benefits of lipid-lowering drugs in "high-risk" individuals who have not experienced a coronary event. These findings represent an exciting advance that emphasizes the value of preventive efforts in curbing CHD. PMID- 10682163 TI - The management of hypercholesterolemia in patients with coronary artery disease: guidelines for primary care. AB - More than 10 million individuals in the United States currently have symptomatic coronary artery disease (CAD). Asymptomatic CAD is even more prevalent. CAD in the United States is responsible for approximately 1.5 million myocardial infarctions, 500,000 deaths, and a total economic burden in excess of $120 billion annually. Fortunately, CAD is preventable in many individuals. Our understanding of CAD has steadily progressed throughout the 20th century, and now several lines of evidence support the importance of cholesterol in both the genesis and management of coronary atherosclerosis. Following identification of the presence of cholesterol in atheromas, Anitschkov early this century demonstrated that atherosclerotic lesions can be induced in susceptible animals by high-saturated-fat and cholesterol diets. These lesions regressed when low-fat and cholesterol diets were resumed. In the 1970s and 1980s, findings from the landmark Framingham Heart, Seven Countries, and Multiple Risk Factor Intervention Trial studies firmly established that hypercholesterolemia was a major risk factor for cardiovascular morbidity and mortality. During the 1980s and 1990s, 21 of 22 angiographic trials demonstrated reduced progression of coronary and/or carotid artery disease using lifestyle, drug, and surgical means for reducing cholesterol. The later trials commonly employed hydroxymethylglutaryl coenzyme A reductase inhibitors (statins), reflecting increasing clinical use of these drugs. In 1988, the Adult Treatment Panel of the National Cholesterol Education Program (NCEP) published guidelines on testing and treating hypercholesterolemic patients, which outlined a more aggressive approach to cholesterol lowering than was currently in practice. Since 1994, five large cardiovascular event trials and a large angiographic trial have shown that aggressive cholesterol lowering reduces both cardiac morbidity and mortality, largely substantiating the NCEP guidelines. Although important clinical questions remain regarding patient subsets and treatment goals, lifestyle changes and appropriate drug therapy have proved to be highly effective in preventing initial and recurrent cardiovascular events. PMID- 10682164 TI - Pathogenesis of asthma. AB - The conception of the pathogenesis of a disease is probably as important in determining the selection of therapies as is the evidence provided by outcome studies of their efficacy. The recent evolution in our understanding of the pathogenesis of asthma has nicely paralleled advances in clinical research on new forms of treatment. This evolution has occurred so smoothly that we may not be fully aware how far it has taken us. Airflow obstruction is still regarded as the fundamental cause of the characteristic asthmatic symptoms of shortness of breath, chest tightness, and wheezing, while the factors leading to airflow obstruction are still assumed to include spasm of airway smooth muscle, thickening of the airway wall, and inspissation of viscid plugs of mucus in the airway lumen. What is new is the recognition of asthma as a chronic disease of the lower airways associated with characteristic inflammatory changes (involving lymphocytes, mast cells, and eosinophils), and possibly irreversible "remodeling" of the airways (by deposition of collagen and proteoglycans, proliferation and transformation of resident cells, and infiltration with inflammatory cells). This modern conception of asthma differs dramatically from the former perception of the disease as an episodic illness characterized by disturbance of the contractile function of airway smooth muscle. The new interpretation has important implications not just for the development of future therapies based on the inflammatory mechanisms involved in the pathogenesis of asthma, but also for the ways in which current therapies should be used. PMID- 10682165 TI - Diagnosis and periodic assessment of asthma. AB - This chapter reviews the features of the clinical history that indicate a diagnosis of asthma, the approach to establishing the diagnosis, and the tests that may be employed in confirming the diagnosis. Recommendations are provided for when to refer to an asthma specialist for a consultation or for ongoing care. The chapter also describes the goal of therapy in asthma and recommends methods for monitoring asthma, along with specific strategies to improve asthma care and to enhance communication between the health care provider and the patient. Much of the content comes directly from the recent Guidelines for the Diagnosis and Management of Asthma developed by the National Institutes of Health (NIH)-NHLBI National Asthma Education and Prevention Program (1). Other recent NIH publications that may provide a valuable resource for the clinician are listed (2 8). PMID- 10682166 TI - Pharmacologic therapy for asthma. AB - Pharmacologic therapy is used to prevent and control asthma symptoms, reduce the frequency and severity of asthma exacerbations, and reverse airflow obstruction. Recommendations in this chapter, based on the 1997 National Asthma Education and Prevention Program Expert Panel Report II: Guidelines for the Diagnosis and Management of Asthma, reflect the scientific concept that asthma is a chronic disorder with recurrent episodes of airflow limitation, mucus production, and cough. Asthma medications are categorized into two general classes: long-term control medications taken daily on a long-term basis to achieve and maintain control of persistent asthma (these medications are also known as long-term preventive, controller, or maintenance medications), and quick-relief medications taken to provide prompt reversal of acute airflow obstruction and relief of accompanying bronchoconstriction (these drugs are also known as reliever or acute rescue medications). Patients with persistent asthma require both classes of medication. Selecting the appropriate pharmacologic therapy to achieve and maintain control of asthma involves several considerations: the medications and their routes of administration, a stepwise approach to managing asthma long-term as a chronic disorder, and the development of an effective clinician-patient partnership strategy where patient education is continuously provided. PMID- 10682167 TI - Management of asthma exacerbations. AB - The 1997 Expert Panel Report 2 from the National Asthma Education and Prevention Program details principles and goals for managing asthma exacerbations, based on scientific literature and the opinion of the panel. The panel's recommendations are summarized here, along with approaches to the evaluation and management of patients with asthma exacerbations. Methods to assess and classify the severity of asthma exacerbations are discussed, and treatment objectives for mild, moderate, and severe exacerbations are presented, along with a discussion of postinfectious acute airway hyperresponsiveness. A review of pharmacologic agents used in the treatment of asthma exacerbations is also included. Key points in the management of asthma exacerbations include the notion that early treatment is the best strategy for management. Important elements of early treatment include recognition of early signs of worsening asthma, a written action plan to guide patient self-management, appropriate intensification of therapy, and prompt communication between patient and provider about deterioration in asthma control. Other key points include the use of inhaled beta 2-adrenergic agonists to provide prompt relief of airflow obstruction, the early use of systemic corticosteroids for patients with moderate to severe exacerbations or for patients who fail to respond promptly and completely to an inhaled beta 2-adrenergic agonist, and monitoring response to therapy with serial measurements of lung function. PMID- 10682169 TI - New classification and diagnostic criteria for diabetes mellitus. AB - There has been an explosive growth in knowledge about diabetes mellitus since the National Diabetes Data Group promulgated diagnostic criteria and a classification system in 1979 that was largely adopted by the World Health Organization. However, recent findings regarding the levels of glucose associated with development of retinopathy, and growing confusion caused by a system of classification of diabetes based largely on the treatment used have led to a new assessment of the diagnosis and classification of diabetes mellitus. Using new data from population-based studies, and placing emphasis on a pathophysiology based system of classification, in 1997, the Expert Committee of the American Diabetes Association released its recommendations for the classification and diagnosis of diabetes. The major changes from the 1979 report include: (a) the preferred use of the terms "type 1" and "type 2" instead of "insulin-dependent" and "non-insulin-dependent" to designate the two major types of diabetes mellitus; (b) a simplification of the diagnostic test to two fasting plasma glucose (FPG) determinations; and (c) a lower cutoff for FPG (126 mg/dL) to diagnose diabetes (this level of FPG having been found equivalent to the 200 mg/dL value in the oral glucose tolerance test for diagnosis). These changes provide an easier and more reliable means of diagnosing persons at risk of complications of hyperglycemia. Even though the fasting criterion was lowered, the total number of persons who will be diagnosed with diabetes by exclusive reliance on FPG will actually be somewhat less than with the old criteria. Moreover, epidemiologic data support the recommendation that screening for diabetes should start at age 45 and be repeated every 3 years in persons without risk factors, and earlier and more often in those with risk factors. PMID- 10682168 TI - Allergen and irritant control: importance and implementation. AB - The Expert Panel Report 2. Guidelines for the Diagnosis and Management of Asthma (1) begins its section on controlling factors that precipitate or worsen asthma with the statement: "For successful long-term asthma management, it is essential to identify and reduce exposures to relevant allergens and irritants and to control other factors that have been shown to increase asthma symptoms and/or precipitate asthma exacerbations." The presence of allergy to indoor allergens and certain seasonal fungal spores has been found to be a risk factor for asthma in epidemiologic studies around the world. Generally between 70% and 85% of asthmatic populations studied have been reported to have positive skin-prick tests. Exposure of allergic patients to inhalant allergens increases airway inflammation, airway hyper-responsiveness, asthma symptoms, need for medication, severe attacks, and even death due to asthma. Environmental tobacco smoke exposure has been shown to increase the prevalence of childhood asthma and to increase asthma symptoms and bronchial hyperresponsiveness while reducing pulmonary function in children chronically exposed. Exposure to other indoor irritants, largely products of unvented combustion, has also been found to increase asthma symptoms. Outdoor air pollution increases asthma symptoms; levels of specific pollutants correlate with emergency room visits and hospitalization for asthma. Rhinitis/sinusitis and gastroesophageal reflux are commonly associated with asthma, and treatment of these conditions has been shown to improve asthma. In patients sensitive to aspirin and nonsteroidal anti inflammatory drugs or metabisulfites, exposure to these agents can precipitate severe attacks of asthma. Viral infections are common causes for exacerbations of asthma. Infections with Mycoplasma pneumoniae and Chlamydia pneumoniae contribute to acute exacerbations and perhaps to long-term morbidity, as well. This chapter will discuss preventive and therapeutic measures that have been found effective in reducing the impact of aggravating or precipitating factors in patients with asthma. PMID- 10682170 TI - Clinical implications of the insulin resistance syndrome. AB - Insulin resistance syndrome (IRS), also termed syndrome X, is a distinctive constellation of risk factors for the development of type 2 diabetes mellitus and cardiovascular disease. The syndrome's hallmarks are glucose intolerance, hyperinsulinemia, a characteristic dyslipidemia (high triglycerides; low high density lipoprotein cholesterol, and small, dense low-density lipoprotein cholesterol), obesity, upper-body fat distribution, hypertension, and increased prothrombotic and antifibrinolytic factors. Insulin resistance, caused by a complex of genetic and environmental influences, is now recognized not just as a mechanism contributing to hyperglycemia in type 2 diabetes, but also as an early metabolic abnormality that precedes the development of overt diabetes. The clinical definition of insulin resistance is the impaired ability of insulin (either endogenous or exogenous) to lower blood glucose. In some insulin resistant individuals, insulin secretion will begin to deteriorate under chronic stress (glucose toxicity) and overt diabetes will result. If not, individuals will remain hyperinsulinemic, with perhaps some degree of glucose intolerance, together with other hallmarks of the IRS. The statistical correlation between hypertension and impaired glucose tolerance is clear, although the mechanism is not yet fully understood. Epidemiologic evidence of insulin resistance as an independent risk factor for atherosclerosis and coronary heart disease (CHD) completed the evolving concept of IRS as the common soil for the development of both diabetes and CHD. No single laboratory test exists for diagnosis of IRS. Rather, IRS remains a clinically evident syndrome that can be suspected on the basis of physical and laboratory findings. This identifies individual patients whom the clinician should screen for associated comorbid conditions, aggressively control cardiovascular risk factors, and tailor drug therapy for optimal benefit. This article provides practical guidelines to achieve these goals and specific strategies to ameliorate cardiovascular and metabolic risk in the IRS. PMID- 10682171 TI - Glucose control in type 1 diabetes: from conventional to intensive therapy. AB - The discovery of insulin at the University of Toronto in 1921 and its first administration on January 11, 1922, dramatically revolutionized the management and outcome of type 1 diabetes. Type 1 diabetes was transformed from a condition that was almost uniformly fatal into a chronic disease characterized by the long term microvascular and macrovascular complications of diabetes. The Diabetes Control and Complications Trial (DCCT) (1), following the lead of smaller European studies, demonstrated conclusively that improved glycemic control, as assessed by glycated hemoglobin (Hb A1c), markedly reduced the development and progression of retinopathy, neuropathy, and nephropathy. These benefits were achieved by the implementation of intensive diabetes management regimens (2). Unfortunately, the advantages of intensive diabetes management were also accompanied by an increased rate of severe hypoglycemia and weight gain. Nonetheless, on balance, it is clear that the benefits of intensive therapy far outweigh the risks. Indeed, the concluding recommendation of the DCCT is that most patients with type 1 diabetes should be treated with intensive diabetes management with the goal of achieving the best possible glycemic control. This article will be divided into four sections providing clinically relevant information relating to the transferral of patients from conventional to intensive diabetes management. We will review the principles of intensive diabetes therapy: selection of patients; implementation of therapy; and assessment of outcome measures. PMID- 10682172 TI - Management of glycemia in type 2 diabetes. AB - The management of type 2 diabetes has been revolutionized over the last 3 to 5 years as a result of dramatic changes in our health care system, new clinical trial data, novel pharmacologic agents, and a better understanding of appropriate methods for patient education regarding lifestyle issues. As a result, diabetes management has become much more heterogeneous, with dramatic differences in style and approach used by practitioners, whether diabetes specialists, primary care providers, or allied health professionals. Diabetes care has also become much more rewarding: The vast majority of patients can now achieve excellent glycemic control while leading full and unrestricted lives. In this article, a construct is reviewed by which comprehensive diabetes care may be approached in a primary care setting. The overall goal of diabetes management should be to provide an opportunity for patients to live out their normal life expectancies with minimal complications. In other articles, screening and treatment suggestions for diabetes complications are provided. Prospective interventional and epidemiologic studies demonstrate that glycemic control is critical to avoidance of complications; the methods to achieve glycemic control are the major focus of this chapter. PMID- 10682173 TI - Prevention of diabetes complications. AB - Basic and clinical research findings have led to an increased understanding about diabetes and its complications. Therapeutic approaches are now based not only on predicted effects from epidemiologic, correlative, or retrospective analyses, but often on prospective intervention trials comparing a new form of therapy to the standard methods. While this database may never be complete, partially due to the complexity and variability of the diabetic state, we now have excellent data that allow the development of aggressive new guidelines for care. Much of the material presented here reflects the views of the American Diabetes Association, as included in a recent publication (1). These guidelines are under review by a number of other organizations and will be subject to modification for special situations. Thus, the terminology "guidelines," rather than "standards of care," is chosen to indicate the flexibility necessary in developing such recommendations for general usage. PMID- 10682174 TI - Clinical aspects of depression. AB - In recent years, clinicians and epidemiologists have examined the differences between depressed patients in primary care and psychiatric settings. Although evidence to support antidepressant efficacy is largely derived from studies of major depression, many patients in primary care settings fall into "nonmajor" depression diagnostic categories. In deciding when to initiate treatment, functional change may be even more important than discrete symptom profiles. Recognizing and treating depression as a comorbid condition in patients with other medical illnesses represents an additional challenge for the primary care physician. Variations in the clinical presentation of depression based on gender, age, culture, or personality must also be considered. PMID- 10682175 TI - Basic neuropharmacology of antidepressants relevant to the pharmacotherapy of depression. AB - In the past decade, 8 new antidepressants were approved for use in the United States by the US Food and Drug Administration. Six of these have been marketed in the United States since 1992. Two additional drugs marketed outside the United States as antidepressants have been approved in this country for obsessive compulsive disorder. Together with the conventional antidepressants, the new ones, and the 2 drugs available off-label, the physician can choose from a list of 22 compounds to treat the depressed patient. This article reviews theories about the mechanisms of action of antidepressants and the basic pharmacology of the newer-generation drugs in relation to the older compounds. We hope this information is of use to the clinician in treating acute depression with pharmacologic agents. PMID- 10682176 TI - Antidepressant options in primary care. AB - This article summarizes the basic and clinical pharmacology of the available antidepressants to aid the busy primary care practitioner in anticipating and predicting the effects of different antidepressants on patients; choosing a specific antidepressant for each patient based on these effects; selecting a sequential antidepressant for the patient who has not benefited from a trial of a specific type of antidepressant, either because of inadequate effect or treatment limiting adverse effects; and avoiding adverse drug interactions when choosing an antidepressant for patients on other drug therapy. PMID- 10682177 TI - Strategies for treatment-resistant depression. AB - Clinical depression is a common disorder with serious implications that is often misdiagnosed or underestimated. More than 50% of suicides are associated with a major depressive disorder, and depression can adversely impact a variety of other medical conditions. Although > or = 70% of depressions can respond to appropriate medications, few patients actually receive adequate medical therapy. This article reviews the definition of adequate therapy for depression, discusses common comorbid conditions, and examines the issue of true treatment resistance. Practical strategies for treatment-resistant depression, including switching classes of antidepressant drugs, combination therapy, augmentation strategies, and somatic therapies, are incorporated into a treatment algorithm. PMID- 10682178 TI - Gastroesophageal reflux: practical management of a common, challenging disorder. AB - Gastroesophageal reflux (GER) occurs in 2 distinct forms that differ in pathophysiology, clinical presentation, natural history, and therapy: mild GER (with no or minimal esophagitis) and classic, severe reflux (at risk for erosive esophagitis). A minority of subjects (< 20%) have the classic, potentially severe pattern of GER caused by reduced lower esophageal sphincter (LES) pressure and prolonged acid reflux, particularly at night, but also during the day. Evaluation and management must be catered to patients with this pattern of reflux. In contrast, symptoms in mild reflux (the majority) often occur during the day after meals in an upright posture (upright reflux); resting LES pressure is usually normal (reflux episodes are related to transient relaxation of the LES) and little reflux occurs at night. Acid reflux, which occurs mostly during the day, overlaps with the normal range and esophagitis is rare; however, symptoms can be distressing. Optimal management is controversial because no outcome trials have been conducted to address management in primary care settings. However, clinical clues can help differentiate mild and severe reflux and guide management decisions. This article provides a detailed approach to current management of GER syndromes. PMID- 10682179 TI - Diagnosis and management of Helicobacter pylori infection. AB - Helicobacter pylori infects more than half of the world's population, making it one of the most prevalent infections. H pylori is now accepted as the most common cause of histologic gastritis and is responsible for the majority of cases of peptic ulcer disease and gastric cancer. Approximately 1 in 6 (17%) persons with H pylori infection will develop peptic ulcer disease, and each year 1% to 2% of these will experience a major or life-threatening complication, such as bleeding, perforation, or gastric outlet obstruction. Peptic ulcer disease should no longer be regarded as a chronic, recurring, lifelong disease, but rather as a curable infectious disease. The diagnosis and therapy of this common infectious disorder have become increasingly straightforward. In this article, we discuss the possible outcomes of long-standing infection, the various diagnostic tests available, including whom and when to test, and the combination drug regimens approved for cure. PMID- 10682180 TI - Peptic ulcer and dyspepsia. AB - Peptic ulcers are defects in the gastrointestinal mucosa that extend through the muscularis mucosae. They persist as a function of the acid or peptic activity in gastric juice. Twenty years ago, most ulcers were considered idiopathic; but a revolution in knowledge has occurred, so that it is now understood that the great majority of ulcers results from infection with Helicobacter pylori (HP) or use of nonsteroidal anti-inflammatory drugs (NSAIDs). Before this revolution, peptic ulcer disease was a common public health problem, responsible for considerable morbidity, some mortality, and high economic cost. Today, the overall prevalence of ulcers is falling, but complication rates remain relatively stable. These complex trends primarily reflect 3 factors: the rapid decline in the prevalence of HP in the population of developed countries, an increase in consumption of NSAIDs, and change in rates of smoking. Peptic ulcer prevalence is falling in younger individuals because of decreased prevalence of HP, whereas complications are rising in older subjects, largely as the result of increased NSAID use. PMID- 10682181 TI - NSAID-related gastrointestinal complications. AB - Despite the common induction of gastrointestinal (GI) complications by nonsteroidal anti-inflammatory drugs (NSAIDs), many aspects of pathogenesis and management remain controversial. The most important complications are bleeding and perforation arising in the esophagus, stomach, and duodenum due to NSAID effects on platelets and on a variety of mucosal lesions. Complications arise from preexisting peptic ulcer, NSAID-induced ulcers and erosions, and other lesions (not caused by NSAIDs) caused to bleed by NSAID-induced platelet dysfunction. Much confusion has arisen from the umbrella term "NSAID gastropathy" used to embrace a variety of pathogenetically distinct mucosal lesions. Failure to discriminate among the different forms of NSAID injury has hampered clinical investigation. This article offers general guidelines for prevention of NSAID complications, but there remain many unresolved issues, including the role of Helicobacter pylori infection. The best approach to management is to remove or reduce exposure to NSAIDs whenever possible. The newer NSAIDs (e.g., cyclo oxygenase [COX]-2-selective inhibitors) have a much lower risk of endoscopic ulcers and minimal platelet effects, which are likely to translate into lower risk of GI complications. However, this benefit on clinical outcome remains to be established. PMID- 10682182 TI - Common functional gastrointestinal disorders: nonulcer dyspepsia and irritable bowel syndrome. AB - Up to 35% of the world population suffer from functional gastrointestinal disorders (FGD), accounting for about 40% of gastroenterologic and 12% of primary care practice. Society incurs high costs from FGD morbidity in terms of medical workups and absenteeism from work. FGD are characterized by chronic and recurrent symptoms of the gastrointestinal (GI) tract without detectable structural or biochemical abnormalities. In the absence of universal biologic markers, the diagnosis is based on consensus symptom criteria (1). This chapter reviews current knowledge of the pathophysiology and provides a practical approach to patients with functional dyspepsia and irritable bowel syndrome, 2 of the most common functional GI syndromes. PMID- 10682183 TI - Classification and epidemiology of headache. AB - Headache disorders are remarkably common. Like back pain, headache is a symptom that has a broad range of possible causes. Diagnosis of primary headache disorders (migraine, tension-type headache, cluster headache) depends on systematic exclusion of secondary disorders and systematic identification of the specific features of the primary disorders. Thus, migraine should be viewed as an episodic syndrome of pain, involving intracranial structures associated with other neurologic disturbances. Because of the large number of potential etiologies, clinicians must approach headache classification systematically. In this chapter, we provide an overview of headache classification followed by discussions of epidemiology. PMID- 10682185 TI - Diagnosis and treatment of migraine. AB - Migraine is one of the most common and misunderstood disease encountered in general medical practice. An estimated 23 million Americans suffer disabling migraines, yet only a minority are diagnosed (1,2). An even smaller percentage receive optimal care. Migraine extracts a significant personal, psychologic, social, and economic toll from migraineurs and their families. An estimated 150 million workdays are lost annually due to headache at an estimated cost of $6 to $17 billion (3,4). Recent advances in understanding of the pathophysiology and acute therapy provide the potential to markedly reduce the impact of migraine. Available abortive medications have efficacy rates as high as 80%, but only a minority of afflicted patients currently receive these therapies. While reducing headache pain, they also restore function, enabling an individual to return to work, family, and personal commitments (5). Future progress in migraine management resides in early identification and optimization of migraine treatment. This article focuses on diagnosis and treatment. PMID- 10682184 TI - Headache diagnosis. AB - Headache is an extremely common symptom in primary care practice. Despite the ubiquity of the pain, differential diagnosis of headache is not difficult, provided the clinician obtains a comprehensive history. A complete physical examination and specific testing may be required to rule out other underlying causes, but headache itself falls into 3 main classes that are readily identified. Migraine headache occurs most commonly in women, is of moderate to severe intensity, and is often accompanied by nausea and increased sensitivity to light and sound. Cluster headache describes multiple recurrent attacks of severe unilateral pain and occurs most frequently in men. Tension-type headache is the most common form, characterized by mild to moderate dull pain that is often brought on by stress and/or depression. Understanding the triggers and manifestations of these headache types is essential for effective management. PMID- 10682186 TI - Tension-type headache. AB - Tension-type headaches, the most prevalent form of headache, are differentiated as being either episodic or chronic. The episodic form is a physiologic response to stress, anxiety, depression, emotional conflicts, fatigue, or repressed hostility. Treatment focuses on the use of over-the-counter or prescribed simple analgesics for pain relief. Successful treatment of the chronic form depends on recognition of depression or persistent anxiety states. Primary care physicians can effectively manage most of these patients with nonhabituating anxiolytic or antidepressant medications; however, referrals for psychotherapy may be required in some cases. When tension-type headaches occur in children and adolescents, the physician must explore the patient's family and social relationships as well as school performance. In addition to nonhabituating drug therapies, family counseling and biofeedback may be helpful. In coexisting migraine and tension type headaches, nonhabituating analgesics may be used for the relief of acute pain; the use of ergotamine and triptans should be restricted to relief of the hard or sick headache. Tricyclic antidepressants or monoamine oxidase inhibitors are the gold standards for prophylaxis, although the selective serotonin reuptake inhibitors may be indicated in less severe cases. Several forms of biofeedback have also proved effective. Nonetheless, some patients with this form of headache may require psychiatric treatment for severe depression. PMID- 10682187 TI - Emergency department management of the acute headache. AB - Headache is a common complaint of patients seeking care at an emergency department (ED). A survey of more than 16,755 walk-in patients at an ED showed that 323 (1.9%) had a chief complaint of migraine (1). Almost one sixth of these patients had used the ED more than once. In fact, migraineurs used the ED and other health care providers 2 to 5 times more than nonmigraineurs (2). Fortunately, headaches associated with significant morbidity and mortality occur infrequently (3). The ED physician must be able to address the patient's need for pain management and establish the correct diagnosis for the headache while also ruling out any possibility of organic disease or life-threatening illness. Potential problems include ensuring appropriate follow-up and avoidance of narcotic habituation. PMID- 10682189 TI - Moderate alcohol intake protective effect PMID- 10682188 TI - Reduced mortality with moderate alcohol intake PMID- 10682190 TI - Broader use of lipid-lowering therapy argued. PMID- 10682191 TI - The diabetic patient as paradigm for selective antihypertensive therapy. AB - General recommendations from US and international organizations indicate that an ideal approach to the therapy of hypertension should begin with lifestyle modifications, such as decreased salt and fat intake and a careful aerobic exercise program, with the therapeutic goal of a blood pressure (BP) < 140/90 mm Hg. The most recent guidelines recommend more rigorous targets for BP lowering in high-risk populations, such as those with hypertension and concomitant diabetes and/or renal disease with proteinuria. This chapter addresses hypertension in patients with diabetes as an example of a group at especially high risk. It reviews recent clinical trials that support more rigorous BP goals in such patients to reduce cardiovascular morbidity and mortality and considers the importance of combination therapy in achieving these goals. PMID- 10682192 TI - Does the evidence from clinical trials justify the treatment of hypertension? AB - Hypertension is the most common reason for physician office visits. An estimated 43 to 50 million people in the United States have elevated blood pressure (BP). The prevalence of hypertension varies from a small percentage among individuals in their teens and 20s to more than 70% of the elderly (those > 75 years of age). The US Public Health Services (USPHS) has set a goal of having 50% of individuals with hypertension "under control"--that is, reducing their BP to < 140 mm Hg systolic and < 90 mm Hg diastolic--by the year 2000. Current information suggests that we will fall far short of reaching that goal. In the early 1990s, such levels had been achieved by only 27% of hypertensive Americans 18 to 74 years old and by only 20% of those > 75 years old. Is this a cause for concern? Is the goal defined by the USPHS justified? Should we push even harder to reach that goal, should we strive to do even better, or should we be satisfied? Since we are now in the era of evidence-based medicine, the only way to answer these questions is to review what we have learned from the many large, prospective, well-controlled, long-term clinical trials that have addressed these questions over the past 3 decades. PMID- 10682193 TI - Diagnosis and evaluation of secondary hypertension. AB - Although patients with secondary hypertension comprise only a small percentage of those with elevated blood pressure, this subgroup should not be ignored. In many cases, correcting the cause of secondary hypertension can lead to a cure, thus avoiding the need for long-term medical therapy, with its attendant risks and economic toll. Moreover, effective treatment of secondary hypertension can prevent chronic complications, such as left ventricular hypertrophy and coronary artery disease, which markedly increase morbidity and mortality. Nearly all forms of secondary hypertension are related to decreased renal function and/or derangement of hormonal balance or secretion. If hypertension is secondary to chronic renal failure of any etiology, it can be recognized from biochemical assays for blood urea nitrogen and creatinine. PMID- 10682194 TI - Diagnosis and management of hypertensive emergencies. AB - It has been estimated that approximately 600,000 to 800,000 Americans will develop a hypertensive crisis (Calhoun and Oparil, 1990). Although such numbers represent only about 1% of the estimated 60 million Americans with hypertension, hypertensive crisis often constitutes a major medical emergency, necessitating a focused, assertive, and reasoned therapeutic intervention. When such patients are seen in the emergency department or in a physician's office with a critical elevation in blood pressure (BP), appropriate and efficacious management is essential to avoid catastrophic injury to vital target organs, including the central nervous system, the heart, and the kidneys. Delays in initiating effective therapy or, equally important, overzealous therapy leading to a too rapid reduction in BP can produce severe complications involving these target organs. This article reviews the spectrum of clinical syndromes that comprise hypertensive emergencies, highlighting 2 to illustrate the complexities of clinical presentation and management. The newly advocated treatment guidelines based on the category of acute severe hypertension (including asymptomatic hypertensive urgencies) are also considered, as are therapeutic strategies utilizing currently available antihypertensive agents. PMID- 10682195 TI - Broadening behavioral decision research: multiple levels of cognitive processing. AB - The area of behavioral decision research--specifically, the work on heuristics and biases--has had a tremendous influence on basic research, applied research, and application over the last 25 years. Its unique juxtaposition against economics has provided important benefits, but at the cost of leaving it disconnected from too much of psychology. This paper explores an expanded definition of behavioral decision research through the consideration of multiple levels of cognitive processing. Rather than being limited to how decision makers depart from optimality, we offer a broader analysis of how decision makers define the decision problem and link decisions to goals, as well as a more detailed focus on processes associated with implementing decisions. PMID- 10682196 TI - Changing plans: dynamic inconsistency and the effect of experience on the reference point. AB - This article presents a new test of a principle of decision making called dynamic consistency. This principle was tested in an experiment in which participants were asked to make decisions about a second gamble within a sequence of two gambles. Participants were first asked to make a planned choice about the second gamble. The planned choice was made before the first gamble was played and was conditioned on the anticipated outcomes of the first gamble. After the first gamble was played, the same participants were asked to make a final choice about the second gamble, conditioned on the experienced outcome of the first gamble. The results showed that participants' final choices were frequently inconsistent with their plans, even when the anticipated and experienced outcomes were identical. These inconsistencies occurred in a systematic direction. Experiencing an anticipated gain resulted with a change toward risk aversion, and experiencing an anticipated loss resulted in a change toward risk seeking. These results are explained in terms of the effect of actual experience on the reference point used for the evaluation of the decision problem. PMID- 10682197 TI - Value seeking and prediction-decision inconsistency: why don't people take what they predict they'll like the most? AB - In this research, it is proposed that, when making a choice between consumption goods, people do not just think about which option will deliver the highest consumption utility but also think about which choice is most consistent with rationales--beliefs about how they should make decisions. The present article examines a specific rationale, value seeking. The value-seeking rationale refers to the belief that one should choose the option in a choice set that has the highest monetary value. Studies 1 and 2 show that value seeking could lead to a prediction-decision inconsistency, predicting a high consumption utility from one option but choosing another option. Study 3 shows that the prediction-decision inconsistency could be created even by "illusory" (as opposed to truly monetary) values and that the inconsistency could be turned on or off through empirical manipulation. PMID- 10682198 TI - The semantic side of decision making. AB - The research reported in this paper follows the perspective that decision making is a meaningful act that conveys information. Furthermore, the potential meanings associated with decision options may affect the decisions themselves. This idea is examined in the contexts of compensation, donation, and exchange. In general, judgments were relation dependent and meaning dependent. Furthermore, the results show nonmonotonicities and limited substitutability in a pattern that challenges straightforward ways of mapping decisions onto a common currency of utility. PMID- 10682199 TI - Counterfactual thinking and decision making. AB - Recent research on counterfactual thinking is discussed in terms of its implications for decision making. Against a backdrop of the functional benefits of counterfactual thinking, two distinct types of bias, one liberal and one conservative, are discussed. Counterfactuals may cause decision makers to become liberally biased (i.e., capricious) in terms of tactics, but conservatively biased (i.e., rigid) in terms of long-term strategy. That is, counterfactuals may lead to short-term corrective changes that are needless and costly, but they may also lead to long-term overconfidence, blinding the decision maker to possible beneficial strategic adjustments. Recent research on counterfactual thinking, which is inherently multidisciplinary, is reviewed in light of a theoretical structure that posits two mechanisms by which counterfactual effects occur: contrast effects and causal inferences. PMID- 10682200 TI - Choosing remedies after accidents: counterfactual thoughts and the focus on fixing "human error". AB - The present research is motivated by an interest in why organizational decision makers so often respond to accidents with remedy plans that focus narrowly on correcting human error rather than more environment-focused plans or more encompassing plans. We investigated the role of counterfactual thinking in the decision-making tendency toward human-focused plans. Our experiments indicated that even in a domain where human-focused remedies were not otherwise appealing, many participants decided on human-focused remedies after they had generated an "if only" conjecture about the accident. This reflects that human actions are often selected as the focus of "if only" conjectures and, importantly, that this focus "locks in" and carries through to subsequent remedy decisions. Our hypothesis that remedy plans are produced from "if only" thoughts was supported over several alternative interpretations. We discuss implications for research on the relation between counterfactual thinking and adaptive learning. PMID- 10682201 TI - Analogical encoding facilitates knowledge transfer in negotiation. AB - Information learned in one situation often fails to transfer to a similarly structured situation. However, prior findings suggest that comparing two or more instances that embody the same principle can promote abstraction of a schema that can be transferred to new situations. In two lines of research, we examined the effects of analogical encoding on knowledge transfer in negotiation situations. In Experiment 1, undergraduates were more likely to propose optimal negotiation strategies and less likely to propose compromises (a suboptimal strategy) when they received analogy training. In Experiment 2, graduate management students who drew an analogy from two cases were nearly three times more likely to incorporate the strategy from the training cases into their negotiations than were students given the same cases separately. For both novices and experienced participants, the comparison process can be an efficient means of abstracting principles for later application. PMID- 10682202 TI - Why some are perceived as more confident and more insecure, more reckless and more cautious, more trusting and more suspicious, than others: enriched and impoverished options in social judgment. AB - In line with the principle of compatibility, when making social judgments, people tend to focus on personality attributes compatible with the trait under consideration. Better known, or enriched, personages are more likely to present attributes that are compatible with a particular trait than are personages about whom little is known. As a result, enriched personages are more likely to have various, sometimes even conflicting, traits attributed to them. This hypothesis is supported by a number of studies that compare the frequency with which some people are chosen as being better described by opposite trait adjectives than are others. Celebrities more often have both of a pair of opposing adjectives ascribed to them than do less well known figures. Similarly, subjects judge themselves to be better described by either of a pair of opposite adjectives than is a person who is relatively unknown in their lives. The implications for social judgment and for everyday decisions are discussed. PMID- 10682203 TI - Models and mosaics: investigating cross-cultural differences in risk perception and risk preference. AB - In this article, we describe a multistudy project designed to explain observed cross-national differences in risk taking between respondents from the People's Republic of China and the United States. Using this example, we develop the following recommendations for cross-cultural investigations. First, like all psychological research, cross-cultural studies should be model based. Investigators should commit themselves to a model of the behavior under study that explicitly specifies possible causal constructs or variables hypothesized to influence the behavior, as well as the relationship between those variables, and allows for individual, group, or cultural differences in the value of these variables or in the relationship between them. This moves the focus from a simple demonstration of cross-national differences toward a prediction of the behavior, including its cross-national variation. Ideally, the causal construct hypothesized and shown to differ between cultures should be demonstrated to serve as a moderator or a mediator between culture and observed behavioral differences. Second, investigators should look for converging evidence for hypothesized cultural effects on behavior by looking at multiple dependent variables and using multiple methodological approaches. Thus, the data collection that will allow for the establishment of conclusive causal connections between a cultural variable and some target behavior can be compared with the creation of a mosaic. PMID- 10682204 TI - Associative competition in operant conditioning: blocking the response-reinforcer association. AB - Naive rats were trained to leverpress with a 30-sec delay-of-reinforcement contingency from the start of training. In Experiment 1, the delay interval for different groups of subjects included a signal in the first 5 sec, a signal in the last 5 sec, or no signal at any time. Rats with the signal at the start of the delay interval learned most rapidly. Rats with the signal at the end of the delay failed to learn. In Experiment 2, a choice procedure was used, in which each of two levers was associated with its own 30-sec delay of reinforcement. The delay for one lever included a 5-sec signal at the end of the delay. The delay for the second lever had no signal in its 30-sec delay. Preference was in favor of the lever without the signal in the delay interval. The results demonstrate that the acquisition of new response can be blocked in a manner analogous to the blocking of Pavlovian conditioning. PMID- 10682205 TI - The time course of phonological, semantic, and orthographic coding in reading: evidence from the fast-priming technique. AB - The present experiment employed the fast-priming paradigm in reading (Sereno & Rayner, 1992), in which sentences are silently read while eye-movement-contingent changes are made on a specified target region. In this paradigm, when readers fixate on a specified target word region, a prime word is encountered for a brief duration at the beginning of the fixation and then it is replaced by a target word. Three types of primes were employed: homophones, semantically related, and orthographically similar, and five prime durations were employed: 29, 32, 35, 38, and 41 msec. The primary finding was that significant homophone priming was obtained at prime durations ranging from 29 to 35 msec, whereas significant semantic priming occurred only at the 32-msec prime duration. In contrast, significant orthographic priming occurred at all prime durations. These findings indicate that phonological codes are activated during an eye fixation at least as rapidly as semantic codes. An explanation for the pattern of events is suggested using the framework of an activation-verification model. PMID- 10682206 TI - Syntactic priming in written production: evidence for rapid decay. AB - There is strong evidence for syntactic priming in language production (Bock, 1986), but little evidence about the time course of such effects. We report an experiment that examined the circumstances under which syntactic priming decays in written language production. Participants completed sentence fragments that allowed completions with one of two syntactic forms (Pickering & Branigan, 1998). They tended to produce the same syntactic form for immediately consecutive fragments, even though the two fragments described different events. However, when the experimental fragments were separated by other fragments with unrelated syntactic forms, this tendency rapidly diminished. The results suggest that priming effects in written production decay rapidly when other structures are subsequently produced. We discuss the implications for the application of syntactic information during production. PMID- 10682207 TI - Name that tune: identifying popular recordings from brief excerpts. AB - We tested listeners' ability to identify brief excerpts from popular recordings. Listeners were required to match 200- or 100-msec excerpts with the song titles and artists. Performance was well above chance levels for 200-msec excerpts and poorer but still better than chance for 100-msec excerpts. Performance fell to chance levels when dynamic (time-varying) information was disrupted by playing the 100-msec excerpts backward and when high-frequency information was omitted from the 100-msec excerpts; performance was unaffected by the removal of low frequency information. In sum, successful identification required the presence of dynamic, high-frequency spectral information. PMID- 10682208 TI - Figural aftereffects in the perception of faces. AB - We examined figural aftereffects in images of human faces, for which changes in configuration are highly discriminable. Observers either matched or rated faces before or after viewing distorted images of faces. Prior adaptation strongly biases face perception by causing the original face to appear distorted in a direction opposite to the adapting distortion. Aftereffects transferred across different faces and were similar for upright or inverted faces, but were weaker when the adapting and test faces had different orientations (e.g., adapt inverted and test upright). Thus the aftereffects depend on which images are distorted, and not simply on the type of distortion introduced. We further show that the aftereffects are asymmetric, for adapting to the original face has little effect on the perception of a distorted face. This asymmetry suggests that adaptation may play an important normalizing role in face perception. Our results suggest that in normal viewing, figural aftereffects may strongly influence form perception and could provide a novel method for probing properties of human face perception. PMID- 10682209 TI - Recognition memory for faces: when familiarity supports associative recognition judgments. AB - Recognition memory for single items can be dissociated from recognition memory for the associations between items. For example, recognition tests for single words produce curvilinear receiver operating characteristics (ROCs), but associative recognition tests for word pairs produce linear ROCs. These dissociations are consistent with dual-process theories of recognition and suggest that associative recognition relies on recollection but that item recognition relies on a combination of recollection and assessments of familiarity. In the present study, we examined associative recognition ROCs for facial stimuli by manipulating the central and external features, in order to determine whether linear ROCs would be observed for stimuli other than arbitrary word pairs. When the faces were presented upright, familiarity estimates were significantly above zero, and the associative ROCs were curvilinear, suggesting that familiarity contributed to associative judgments. However, presenting the faces upside down effectively eliminated the contribution of familiarity to associative recognition, and the ROCs were linear. The results suggest that familiarity can support associative recognition judgments, if the associated components are encoded as a coherent gestalt, as in upright faces. PMID- 10682210 TI - The importance of monitoring and self-regulation during multitrial learning. AB - Theory suggests that accuracy of metacognitive monitoring and self-regulation of study will affect test performance, but there is little empirical evidence linking these variables. I examined the relation among these variables in a multitrial learning task. Regression analyses showed that monitoring accuracy and self-regulation were reliably related to test performance--greater monitoring accuracy and more effective self-regulation were associated with greater test performance. These analyses were contrasted with analyses typically conducted in previous research, to show the importance of using a multitrial learning task and of attending to the theoretically based causal relation among variables when evaluating how monitoring accuracy and self-regulation are related to test performance. The results of this investigation may help to explain why previous research has failed to link these variables. PMID- 10682211 TI - Aptness predicts preference for metaphors or similes, as well as recall bias. AB - Why might we sometimes prefer a metaphor such as "genes are blueprints" to a simile such as "genes are like blueprints"? One possibility is that metaphors are preferred when the comparison between a tenor (e.g., genes) and a vehicle (e.g., blueprints) seems especially apt. That is, metaphors might be used when the comparison captures many salient features of the tenor in question. The present experiments examined the relation between the aptness of comparisons and people's preferences for expressing those comparisons as metaphors or as similes. In Experiment 1, it was found that there is consensus on how to express particular comparisons. In Experiment 2, it was found that this preference can be predicted from the aptness of a comparison. It was also found that aptness can predict errors in the recall of comparisons. These findings have implications for theories of metaphor. PMID- 10682212 TI - Predicting conjunction typicalities by component typicalities. AB - In two studies, we investigated to what extent typicalities in conjunctive concepts phrased as relative clauses--such as pets that are also birds--can be predicted from simple functions of constituent typicalities and from extensions of such functions. In a first study, analyses of a large aggregated data set, based on seven different experiments, showed that a calibrated minimum rule model and some extensions of this model accounted for a very large part of the variance in the conjunction typicalities. The same models can also account for the so called guppy effect. A psychological explanation is presented, which states that typicalities in contrast categories, like pets that are not birds and birds that are not pets, further improve the prediction of conjunction typicalities. This hypothesis is tested in a second study. PMID- 10682213 TI - Neuroimaging research and the neurobiology of obsessive-compulsive disorder: where do we go from here? PMID- 10682214 TI - Genetics and etiology of schizophrenia and bipolar disorder. PMID- 10682215 TI - Proton spectroscopic imaging of the thalamus in treatment-naive pediatric obsessive-compulsive disorder. AB - BACKGROUND: Neurobiological abnormalities in the thalamus, particularly the dorsomedial nucleus of the thalamus, are believed to be involved in the pathophysiology of obsessive-compulsive disorder. Although obsessive-compulsive disorder commonly arises in childhood and adolescence, no prior study has examined the thalamus in pediatric obsessive-compulsive disorder patients. METHODS: In this study, N-acetyl-aspartate, a putative marker of neuronal viability, creatine/phosphocreatine, and choline levels were measured in the lateral and medical subregions of the left and right thalami using a multislice proton magnetic resonance spectroscopic imaging sequence in 11 treatment-naive, nondepressed obsessive-compulsive disorder outpatients, 8-15 years old, and 11 case-matched control subjects. RESULTS: A significant reduction in N-acetyl aspartate/choline and N-acetyl-aspartate/(creatine/phosphocreatine + choline) was observed in both the right and left medial thalami in obsessive-compulsive disorder patients compared with control subjects. The N-acetyl-aspartate/choline and N-acetyl-aspartate/(creatine/phosphocreatine + choline) levels did not differ significantly between case-control pairs in either the left or the right lateral thalamus. Reduction in N-acetyl-aspartate levels in the left medial thalamus was inversely correlated with increased obsessive-compulsive disorder symptom severity. CONCLUSIONS: These findings provide new evidence of localized functional neurochemical marker abnormalities in the thalamus in pediatric obsessive-compulsive disorder. Our results must be considered preliminary, however, given the small sample size. PMID- 10682216 TI - Molecular genetics of Alzheimer's disease. AB - Application of genetic paradigms to Alzheimer's disease (AD) has led to confirmation that genetic factors play a role in this disease. Additionally, researchers now understand that AD is genetically heterogeneous and that some genetic isoforms appear to have similar or related biochemical consequences. Genetic epidemiologic studies indicate that first-degree relatives of AD probands have an age-dependent risk for AD approximately equal to 38% by age 90 years (range 10% to 50%). This incidence strongly suggests that transmission may be more complicated than a simple autosomal dominant trait. Nevertheless, a small proportion of AD cases with unequivocal autosomal dominant transmission have been identified. Studies of these autosomal dominant familial AD (FAD) pedigrees have thus far identified four distinct FAD genes. The beta-amyloid precursor protein (beta APP) gene (on chromosome 21), the presenilin 1 (PS1) gene (on chromosome 14), and the presenilin 2 (PS2) gene (on chromosome 1) gene are all associated with early-onset AD. Missense mutations in these genes cause abnormal beta APP processing with resultant overproduction of A beta 42 peptides. In addition, the epsilon 4 allele of apolipoprotein E (APOE) is associated with a increased risk for late-onset AD. Although attempts to develop symptomatic treatments based on neurotransmitter replacement continue, some laboratories are attempting to design treatments that will modulate production or disposition of A beta peptides. PMID- 10682217 TI - Molecular and cellular mechanisms of cognitive function: implications for psychiatric disorders. AB - Recent studies on the molecular and cellular basis of learning and memory have brought us closer than ever to understanding the mechanisms of synaptic plasticity and their relevance to memory formation. Genetic approaches have played a central role in these new findings because the same mutant mice can be studied with molecular, cellular, circuit, and behavioral tools. Therefore, the results can be used to construct models that cut across levels of analytical complexity, forging connections from the biochemistry of the modified protein to the behavior of the mutant mice. These findings are not only improving our understanding of learning and memory, they are also enriching our understanding of cognitive disorders, such as neurofibromatosis type I. Mechanisms underlying long-term changes in synaptic function are likely to be at the heart of many cognitive and emotional processes in humans. Therefore, molecular and cellular insights into learning and memory undoubtedly will have a profound impact on the understanding and treatment of psychiatric disorders. PMID- 10682218 TI - Schizophrenia: genes and environment. AB - The historical and genetic foundations of our current understanding of schizophrenia are reviewed, as are the present and future directions for research. Genetic epidemiological investigations, including family, twin, and adoption studies have confirmed the contributions of genetic and environmental determinants of schizophrenia. For example, identical twins show average concordance rates of only 50%; rates of 100% would be expected on the basis of genetic equivalence alone. Genetic factors may cause errors in brain development and synaptic connections. A broad range of environmental components may further damage the brain. Biological components may include pregnancy and delivery complications, such as intrauterine fetal hypoxia, infections, and malnutrition. Primarily nonbiological components may include psychosocial stressors, such as residence in an urban area and dysfunctional family communication. It is likely that the environmental factors interact with the genetic liability in a negative manner to produce disorders in the schizophrenic spectrum. Genetic and environmental components of the disorder are examined, as well as their interactions in producing either neurodevelopmental syndromes or schizophrenia itself. The implication of these findings for prevention and treatment are considered. PMID- 10682219 TI - Search for schizophrenia susceptibility genes. AB - Identification of a gene or genes that contribute to the development of schizophrenia, a complex psychiatric disorder, may be possible through genetic linkage analysis. Although to date no single causative gene has been identified, several chromosomal loci have shown positive linkage results and are under investigation as tentative schizophrenia susceptibility loci. Despite such obstacles as locus heterogeneity among sample populations, epistatic inheritance models, and failure to obtain statistical significance in studies, patterns have emerged that focus research efforts on chromosomes 13, 8, 22, and 6 and 10. Initial heterogeneity analyses suggests that identifiable subgroups of the families may not contribute equally to these linkage findings. Findings on several additional chromosomes await further replication. Future progress in the search for schizophrenia susceptibility genes will require collaboration among researchers from both academia and industry. PMID- 10682220 TI - Familial transmission of risk factors in the first-degree relatives of schizophrenic people. AB - Schizophrenia is a complex illness with multiple pathophysiologic factors that contribute to its psychopathology. One strategy to identify these factors is to observe them in isolation from each other, by characterizing their expression in the relatives of schizophrenic probands. By Mendel's second law, each genetic factor should be independently distributed in a sibship, so that each can be observed by itself, uncomplicated by the general problems of the illness. Such independently distributed phenotypes are obviously useful for genetic analyses; however, they can also be considered together, to model how various brain dysfunctions may combine to produce psychoses. In addition to a sensory gating deficit linked to the alpha 7-nicotinic acetylcholine receptor locus, schizophrenics and their families have a number of other deficits, including decreased hippocampal volume on magnetic resonance images and increased plasma levels of the dopamine metabolite homovanillic acid. Although such research is far from complete, a heuristic model combining a sensory gating deficit, decreased hippocampal neuron capacity, and increased dopaminergic neurotransmission is consonant with current understanding of the neuropsychology of schizophrenia. PMID- 10682221 TI - Bipolar illness and schizophrenia as oligogenic diseases: implications for the future. AB - As with most complex inheritance diseases, there are at this time no identified susceptibility genes for schizophrenia, bipolar manic-depressive illness, major depression, childhood autism, and other inherited brain disorders whose manifestations are primarily behavioral. Nonetheless, progress has occurred. Genetic epidemiologic research, based on reliable phenotypic definitions, has demonstrated the heritability of many of these disorders. Genetic linkages and associations have been reported and replicated, although there have been inconsistencies between studies, apparently due to the low statistical power of the samples studied to detect small effects genes. Nonreplications of early linkage reports in manic-depressive illness in the 1980s occurred when new cases developed in the same large families in which the linkage was originally reported, and the newly ill persons had the wrong genetic markers in the linkage region. This appears to have resulted from applying inappropriate analytic assumptions of single-gene dominant inheritance of a rare gene, which implied that new cases must arise from the same ancestral gene within the pedigree. When new cases arose in family members not sharing that chromosomal region, the initial linkage report was proved invalid. Under oligogenic inheritance, on the other hand, susceptibility genes are expected to be common, and have a substantial probability of being brought into the pedigree by persons marrying in. Nonspecific psychopathology genes may exist, shared by schizophrenia and bipolar illness, diagnoses which do not co-aggregate in families. The discovery of susceptibility mutations may be expected. PMID- 10682222 TI - Susceptibility loci for bipolar disorder: overlap with inherited vulnerability to schizophrenia. AB - Genetic epidemiological studies reveal that relatives of bipolar probands are at increased risk for recurrent unipolar, bipolar, and schizoaffective disorders, whereas relatives of probands with schizophrenia are at increased risk for schizophrenia, schizoaffective, and recurrent unipolar disorders. The overlap in familial risk may reflect shared genetic susceptibility. Recent genetic linkage studies have defined confirmed bipolar susceptibility loci for multiple regions of the human genome, including 4p16, 12q24, 18p11.2, 18q22, 21q21, 22q11-13, and Xq26. Studies of schizophrenia kindreds have yielded robust evidence for susceptibility at 18p11.2 and 22q11-13, both of which are implicated in susceptibility to bipolar disorder. Similarly, confirmed schizophrenia vulnerability loci have been mapped, too, for 6p24, 8p, and 13q32. Strong statistical evidence for a 13q32 bipolar susceptibility locus has been reported. Thus, both family and molecular studies of these disorders suggest shared genetic susceptibility. These two groups of disorders may not be as distinct as current nosology suggests. PMID- 10682223 TI - Pharmacogenetics of antipsychotic treatment: lessons learned from clozapine. AB - The reintroduction of clozapine, the prototype of atypical antipsychotics, in the late 1980s has led to significant advances in the pharmacological management of schizophrenia. Since then, there has been a rapid development of novel "atypical" antipsychotic agents that have been pharmacologically modeled, to a certain extent, after their predecessor clozapine. As with all antipsychotics, there is variability among individuals in their response to these "atypical" drugs. Pharmacogenetics can provide a foundation for understanding this interindividual variability in antipsychotic response. This review first provides a rationale for the pharmacogenetic investigation of this variable trait. Studies of pharmacokinetic and pharmacodynamic factors of antipsychotic therapy are considered in the development of this rationale. Next, the molecular genetic techniques used to study this interindividual variation in response are described. This is followed by a review and discussion of the published studies examining genetic factors involved in clozapine response. From this, several recommendations for future pharmacogenetic investigations of antipsychotic response are proposed. Although still in its early stages, psychiatric pharmacogenetics should provide a basis for individualized pharmacotherapy of schizophrenia, and may also lead to the development of newer, more efficacious antipsychotic agents. PMID- 10682224 TI - Multiple antioxidants in the prevention and treatment of Alzheimer disease: analysis of biologic rationale. AB - The etiology of Alzheimer disease (AD) is not well understood; therefore, neither prevention strategies nor long-term effective treatment modalities are available for this disease. Based on laboratory and clinical studies, it appears that reactive oxygen species (ROS) and reactive nitrogen species (RNS) that are generated extracellularly and intracellularly by various mechanisms are among the major intermediary risk factors that initiate and promote neurodegeneration in idiopathic AD. Therefore, multiple antioxidant supplements could be useful in the prevention of AD, and as an adjunct to standard therapy in the treatment of AD. The products of inflammatory reactions such as prostaglandins (PGs; PGE1 and PGA1), free radicals, cytokines, and complement proteins are neurotoxic. Nonsteroidal antiinflammatory drugs (NSAIDs), which inhibit the synthesis of PGs, reduce the rate of deterioration of cognitive functions in patients with advanced AD. Cholinergic drugs are routinely used in the treatment of AD to improve cognitive functions. Therefore, we propose that a combination of multiple antioxidants and NSAIDs may be more beneficial in the prevention of AD, and that this combination taken together with cholinergic drugs may be more effective in the treatment of AD than the individual agents alone. We also hypothesize that, in idiopathic AD, epigenetic components of neurons such as mitochondria, membranes, other membranous structures, and protein modifications--rather than the genes of neurons--are the primary targets for the action of neurotoxins including free radicals. In some familial AD, mutations in amyloid precursor protein and presenilins are associated with the risk of early onset of this disease; however, their mechanisms of action are not fully understood. PMID- 10682225 TI - Combined electroconvulsive-clozapine therapy. AB - We reviewed 36 reported psychiatric patients who were treated with a combination of electroconvulsive therapy (ECT) and clozapine. The indication of the ECT clozapine treatment was resistance to classical antipsychotic agents, clozapine, or ECT alone. Sixty-seven percent of the patients benefited from the combined treatment. In most of the patients, the combined treatment was safe and well tolerated. Adverse reactions occurred in 16.6% of the patients and included prolonged ECT-induced seizures (one case), supraventricular (one case) and sinus tachycardia, and blood pressure elevation. It seems that combined ECT-clozapine treatment is effective and safe. This strategy may be a therapeutic option in treatment-resistant patients. PMID- 10682226 TI - Lack of residual sedation following middle-of-the-night zaleplon administration in sleep maintenance insomnia. AB - The present randomized, double-blind, placebo and active-drug controlled, crossover study assessed residual sedation after zaleplon 10 mg, flurazepam 30 mg (as an active control), and placebo, taken during a nocturnal awakening in patients with sleep maintenance insomnia. Twenty-two healthy sleep maintenance insomniacs (11 men; mean age, 42 y) received zaleplon, flurazepam, or placebo after an experimental awakening 3.5 hours after bedtime on two consecutive nights in each of three conditions. Residual sedation was measured with sleep latency testing (5 and 6.5 h postdrug), digit symbol substitution, symbol copying, and subjective sleepiness by visual analog scale, each twice each morning. Zaleplon did not differ from placebo on any measure of residual sedation; flurazepam showed significant sedation on all measures. No residual sedative effects were detected 5 or 6.5 hours after ingestion of zaleplon during the middle of the night by sleep maintenance insomniacs. PMID- 10682227 TI - Multiple-dose pharmacokinetics of selegiline and desmethylselegiline suggest saturable tissue binding. AB - The goal of this study was to examine the multiple-dose pharmacokinetics of selegiline and its metabolites desmethylselegiline, 1-methamphetamine, and 1 amphetamine after oral administration of selegiline HCl. Twelve healthy volunteers received 10 mg of selegiline HCl once daily for 8 days. The pharmacokinetic profiles of selegiline and the metabolites were examined from serum samples for 24 hours (i.e., the dosing interval, tau) on days 1, 4, and 8. The results indicated significant apparent accumulation of selegiline and desmethylselegiline during the 8-day period of selegiline administration. The AUC tau S of selegiline and desmethylselegiline were increased 2.7 fold (p < 0.001) and 1.5 fold (p < 0.001), respectively, from day 1 to day 8. However, the half lives of selegiline (range, 1.5-3.5 h) and desmethylselegiline (range, 3.4-5.3 h) were found to be relatively short. Accordingly, the short half-lives of these compounds failed to predict the apparent accumulation. With both of the 1 amphetamine metabolites of selegiline, steady state was reached by day 4. We suggest that the most likely explanation for the apparent accumulation of selegiline and desmethylselegiline was the saturation of the MAO-B binding sites in tissues, although decreased first-pass metabolism of selegiline cannot be ruled out. The observed increase in selegiline and desmethylselegiline concentrations on multiple dosing is not likely to significantly increase the pharmacodynamic effect or adverse effects of selegiline compared with what has been found after a single 10-mg dose. PMID- 10682228 TI - Diurnal motor variations to repeated doses of levodopa in Parkinson's disease. AB - Patients with Parkinson's disease (PD) in long-term levodopa therapy often complain of worsening of motor symptoms in the afternoon and evening. The pathophysiology of this phenomenon is not known. We evaluated the motor response to repeated doses of levodopa during a 12-hour period in 52 parkinsonian patients (19 de novo, 20 stable, and 13 wearing-off). On the day of the study, all patients received standard doses of levodopa/carbidopa at 8:00 a.m., 12:00 noon, and 4:00 p.m. Motor measurements such as tapping test, walking time, and tremor score, and blood samples for levodopa and 3-O-methyldopa (3OMD) plasma analysis, were performed hourly. Mean motor scores and pharmacokinetic data, evaluated for a period of 3 hours after each levodopa dose, were compared. In de novo patients, we did not observe diurnal changes in motor score, whereas a progressive daytime worsening was visible in stable and wearing-off patients. No significant difference in levodopa pharmacokinetics after each levodopa dose was observed within each patient group, whereas 3OMD plasma levels significant increased with repeated levodopa administrations. However, no significant correlation between motor scores and 3OMD plasma levels was observed, suggesting that the diminishing motor response to afternoon and evening doses of levodopa in patients in long term levodopa therapy does not relate to the pharmacokinetics of the drug. It is possible that this phenomenon may be an expression of the occurrence of tolerance to repeated doses of levodopa. PMID- 10682229 TI - A randomized controlled trial comparing pramipexole with levodopa in early Parkinson's disease: design and methods of the CALM-PD Study. Parkinson Study Group. AB - CALM-PD (Comparison of the agonist pramipexole with levodopa on motor complications of Parkinson's disease) is a randomized, multicenter, double-blind, controlled clinical trial designed to compare the policy of initial treatment of pramipexole with the policy of initial treatment with levodopa in early, symptomatic Parkinson's disease with regard to the development of dopaminergic motor complications. At 22 American and Canadian sites, 301 eligible subjects requiring antiparkinsonian therapy to treat emerging disability were enrolled in CALM-PD and randomized to (i) active pramipexole and placebo levodopa or (ii) placebo pramipexole and active levodopa. Subjects are being evaluated systematically at regular intervals during a 23.5-month period to determine if and when dopaminergic motor complications (wearing off, dyskinesias, "on-off" effects) occur. In addition, quality-of-life outcomes, economic outcomes, and functional imaging outcomes are being assessed in standard fashion with [123I] beta-CIT and SPECT imaging throughout the trial. The study design contains many provisions to approximate routine clinical practice and to produce data about clinical effectiveness, tolerability, and cost to facilitate the evidence-based practice of neurology. PMID- 10682230 TI - Gabapentin treatment for muscle cramps: an open-label trial. AB - To evaluate the efficacy and safety of gabapentin in the treatment of muscle cramps, we engaged an open-label trial with a group of 30 patients with frequent (> 5 cramps/week), stable, long-lasting cramps, associated with different diseases. Gabapentin was effective in reducing the frequency and severity of muscle cramps and associated sleep disturbances (clinical outcome measures) within the first 2 weeks of medication at 600 mg/d. At the 1 month control (mean dosage, 825 +/- 35 mg), almost every patient had responded to treatment and two thirds experienced a total remission of symptoms. After 3 months of therapy (mean dosage, 892 +/- 180 mg), cramps disappeared in 100% of patients and this benefit persisted as long as 6 months. Additionally, we evaluated in 10 patients the Cramps Threshold Frequency (CTF) (neurophysiological outcome measure) before and during gabapentin treatment. Gabapentin significantly increased the CTF, returning it to normal values. With the limitation of an open-label methodology, our clinical and neurophysiologic experience suggests that a gabapentin dose of 600-1200 mg/d would be helpful in the treatment of muscular cramps. PMID- 10682231 TI - Bradykinesia in Huntington's disease. AB - Huntington's disease (HD) is characterized by the presence of hyperkinesias, but bradykinesia is also present in most patients. We studied the motor performance of 18 patients with genetically proven HD (age, 38.5 +/- 10 y; clinical stage, 1.7 +/- 1.7; (CAG) triplet length, 49.2 +/- 6.8 triplets; all but three patients were free from neuroleptics) and compared with a control group (n = 18) and with a typical Parkinson's disease (PD) group (n = 20). Motor study included the four timed tests commonly used for PD: Pronation-supination (PS), finger dexterity (FD), movement between two points (MTP) and walking test (WT). Tests were done at 9 AM. The PD group was studied in "off" condition, with no medication given for 12 hours. The HD group was slower than the controls on all tasks (all tests significant, p < 0.01, Mann-Whitney U test) and even slower than PD group (for FD, p < 0.05). A significant correlation was found between each test and clinical stage (for PS, r = 0.84; for FD, r = 0.75; for MTP, r = 087, and for WT, r = 0.77, Pearson). Severe bradykinesia was present in HD, and motor impairment is related to clinical stage. PMID- 10682232 TI - Asterixis induced by gabapentin. AB - We report the case of a patient with postherpetic neuralgia who developed asterixis while being treated with gabapentin. We discuss the possible mechanism of asterixis in this patient. PMID- 10682233 TI - Central nervous system side effects associated with zolpidem treatment. AB - Zolpidem is one of the newer medications developed for the treatment of insomnia. It is an imidazopyridine agent that is an alternative to the typical sedative hypnotic agents. Zolpidem use is gaining favor because of its efficacy and its side effect profile, which is milder and less problematic than that of the benzodiazepines and barbiturates used to treat insomnia. Still, side effects are not uncommon with zolpidem use. We report a series of cases in which the patients developed delirium, nightmares and hallucinations during treatment with zolpidem. We will review its pharmacology, discuss previous reports of central nervous system side effects, examine the impact of drug interactions with concurrent use of antidepressants, examine gender differences in susceptibility to side effects, and explore the significance of protein binding in producing side effects. PMID- 10682234 TI - Intravenous administration of levodopa ameliorated a refractory akathisia case induced by interferon-alpha. AB - A 28 year-old man with chronic hepatitis B was administered interferon-alpha (5 x 10(6) IU) intramuscularly once a day for 28 days. Eight days after the end of interferon treatment, he showed signs of inability to sit still for ten seconds and walked around constantly. His akathisia symptoms worsened thereafter. Clonazepam, thioridazine, beta-blockers, anticholinergics, and bromocriptine did not ameliorate his akathisia. Two days' administration of levomepromazine 100 mg led him to a coma for 2 days. Intravenous levodopa 25 mg ameliorated his akathisia symptoms drastically. He became completely premorbid 2 weeks after administration of levodopa. The present report illustrates a rare case of refractory akathisia after interferon-alpha treatment and also that levodopa treatment would be theoretically and practically useful in reducing the neurotoxicity associated with interferon-alpha. PMID- 10682235 TI - Resting and maximal heart rates in ectothermic vertebrates. AB - Resting and maximal heart rates (HR) in ectothermic vertebrates are generally lower than those in endotherms and vary by more than an order of magnitude interspecifically. Variation of HR transcends phylogeny and is influenced by numerous factors including temperature, activity, gas exchange, intracardiac shunts, pH, posture, and reflexogenic regulation of blood pressure. The characteristic resting HR is rarely the intrinsic rate of the pacemaker, which is primarily modulated by cholinergic inhibition and adrenergic excitation in most species. Neuropeptides also appear to be involved in cardiac regulation, although their role is not well understood. The principal determinants of resting HR include temperature, metabolic rate and hemodynamic requirements. Maximal HRs generally do not exceed 120 b min-1, but notable exceptions include the heterothermic tuna and small reptiles having HRs in excess of 300 b min-1 at higher body temperatures. Temperature affects the intrinsic pacemaker rate as well as the relative influence of adrenergic and cholinergic modulation. It also influences the evolved capability to increase HR, with maximal cardiac responses matched to preferred body temperatures in some species. Additional factors either facilitate or limit the maximal level of HR, including: (1) characteristics of the pacemaker potential; (2) development of sarcoplasmic reticulum as a calcium store in excitation-contraction coupling; (3) low-resistance coupling of myocardial cells; (4) limitations of force development imposed by rate changes; (5) efficacy of sympathetic modulation; and (6) development of coronary circulation to enhance oxygen delivery to myocardium. In evolutionary terms, both hemodynamic and oxygen requirements appear to have been key selection pressures for rapid cardiac rates. PMID- 10682236 TI - Regulation of cardiac rhythm in hibernating mammals. AB - The dramatic fall in heart rate exhibited by mammals entering hibernation begins before there is any noticeable fall in body temperature. The initial, progressive decrease in heart rate is the result of a cyclic parasympathetic activation that induces skipped beats and regular asystoles as well as slows the even heart beat. As body temperature subsequently falls, the parasympathetic influence is progressively withdrawn and periods of parasympathetic and sympathetic dominance alternate and give rise to regular periods of arrhythmia (tachycardia followed by bradycardia), and occasional long asystoles or periods of highly irregular cardiac activity. Superimposed on this is a vagally-mediated, respiratory sinus arrhythmia that is accentuated in species that breathe episodically. These events give way to a uniform heart rate in deep hibernation at low temperatures where both parasympathetic and sympathetic tone appear absent. The complete absence of tone is not a function of reduced temperature but is reflective of the state of deep, steady state hibernation. The elevation in heart rate that accompanies the onset of arousal is the result of dramatic increases in sympathetic activation that precede any increases in body temperature. As body temperature then rises, sympathetic influence is slowly withdrawn. Arrhythmias are also common during natural arousals or shifts from lower to warmer hibernation temperatures as periods of parasympathetic and sympathetic dominance again alternate en route to re-establishing a steady state in euthermia. The mechanism behind, and the biological significance of, cardiac changes mediated through orchestrated arrhythmias remain unknown. PMID- 10682237 TI - Control and interaction of the cardiovascular and respiratory systems in anuran amphibians. AB - In anuran amphibians, respiratory rhythm is generated within the central nervous system (CNS) and is modulated by chemo- and mechanoreceptors located in the vascular system and within the CNS. The site for central respiratory rhythmogenesis and the role of various neurotransmitters and neuromodulators is described. Ventilatory air flow is generated by a positive pressure, buccal force pump driven by efferent motor output from cranial nerves. The vagus (cranial nerve X) also controls heart rate and pulmocutaneous arterial resistance that, in turn, affect cardiac shunts within the undivided anuran ventricle; however, little is known about the control of central vagal motor outflow to the heart and pulmocutaneous artery. Anatomical evidence indicates a close proximity of the centers responsible for respiratory rhythmogenesis and the vagal motoneurons involved in cardiovascular regulation. Furthermore, anurans in which phasic feedback from chemo- and mechanoreceptors is prevented by artificial ventilation exhibit cardiorespiratory interactions that appear similar to those of conscious animals. These observations indicate interactions between respiratory and cardiovascular centers within the CNS. Thus, like mammals and other air-breathing vertebrates, the cardio-respiratory interactions in anurans result from both feedback and feed-forward mechanisms. PMID- 10682238 TI - Environmental influences on the development of the cardiac system in fish and amphibians. AB - In poikilothermic animals body temperature varies with environmental temperature, and this results in a change in metabolic activity (Q10 of enzymatic reactions typically is around 2-3). Temperature changes also modify gas transport in body fluids. While the diffusion coefficient increases with increasing temperatures, physical solubility and also hemoglobin oxygen affinity decrease. Therefore, an increase in temperature typically requires adjustments in cardiac activity because ventilatory and convectional transport of respiratory gases usually are tightly coupled in adults in order to meet the oxygen demand of body tissues. Hypoxic conditions also provoke adaptations in the central circulatory system, like the hypoxic bradycardia, which has been described for many adult lower vertebrates, combined with an increase in stroke volume and peripheral resistance. In embryos and larvae the situation is much more complicated, because nervous control of the heart is established only late during development, and because the site of gas exchange changes from mainly cutaneous gas exchange during early development to mainly pulmonary or branchial gas exchange in late stages. In addition, recent studies in amphibian and fish embryos and larvae reveal, that at least in very early stages convectional gas transport of the hemoglobin is not essential, which means that in these early stages ventilatory and convectional gas transport are not yet coupled. Accordingly, in early stages of fish and amphibians the central cardiac system often does not respond to hypoxia, although in some species behavioral adaptations indicate that oxygen sensors are functional. If a depression of cardiac activity is observed, it most likely is a direct effect of oxygen deficiency on the cardiac myocytes. Regulated cardiovascular responses to hypoxia appear only in late stages and are similar to those found in adult species. PMID- 10682239 TI - Metabolic and ventilatory responses to hypoxia in two altitudinal populations of the toad, Bufo bankorensis. AB - Effects of hypoxia on resting oxygen consumption (MO2), lung ventilation, and heart rate at different ambient PO2 were compared between lowland and high altitude populations of the toad, Bufo bankorensis. Resting MO2 decreased significantly in mild hypoxia (PO2 = 120 mm Hg) at 10 degrees C and in moderate hypoxia (PO2 = 80 mm Hg) at 25 degrees C in both altitudinal populations; however, resting MO2 did not differ significantly between the two populations. Numbers of lung ventilation periods (VP) and total inspired volume (VL) did not change with PO2 at 10 degrees C, but did increase at moderate and severe hypoxia (40 mm Hg), respectively, at 25 degrees C. Resting heart rates did not change during hypoxia and did not differ between altitude populations. The results suggest (1) the effect of PO2 change on MO2 should be considered in future studies involving transfer of anurans to a different altitude; and (2) the metabolic and ventilatory physiology in B. bankorensis does not compensate for the low temperature and PO2 at high altitude. PMID- 10682240 TI - Genetic, environmental and maternal influences on embryonic cardiac rhythms. AB - The relative roles of an animal's genetic constituents and environmental factors in influencing physiological variables such has heart rate have not been extensively investigated. This paper considers how heart rate patterns in the developing animal can be regulated, and how a combination of 'nature' and 'nurture' may interact to produce discrete patterns of heart rate change during development. The concept of the 'developmental trajectory' is evoked to generate a conceptual framework for how physiological development can be perturbed by environmental factors. Data are provided from three species showing how 'clutch effects' (the fact that siblings perform physiologically much more similarly than non-siblings) can greatly influence the variance observed when collecting data on heart rate during development. Finally, so-called 'maternal effects', which are the influences on embryos of environmental experiences of the parents, are discussed as potentially confounding effects in the study of the genetic basis for physiological patterns of change during development. PMID- 10682241 TI - Biological rhythms in birds--development, insights and perspectives. AB - The aim of this review is to show that probably the internal clock of precocial birds is imprinted in the prenatal period by exogenous factors (zeitgeber). The activity of organ functions occurs early during embryonic development, before this function is ultimately necessary to ensure the survival of the embryo. Prenatal activation of some functional systems may have a training effect on the postnatal efficiency. The development of physiological control systems is influenced by endogenous and exogenous factors during the late prenatal and early postnatal period: epigenetic adaptation processes play an important role in the development of animals; they have acquired characteristics which are innated but not genetically fixed. As a rule, the actual value during the determination period has a very strong influence on the set-point of the system. This will be explained using the example of thermoregulation. It is shown in detail that it seems to be possible to imprint the prenatal development of circadian rhythms by periodic changes of the light-dark cycle but not by rhythmic influence of acoustic signals. Altogether, there are more questions open than solved concerning the perinatal genesis of circadian rhythms in birds. Topics are given for the future research. PMID- 10682242 TI - Long-term telemetry of heart rates and energy metabolic rate during the diurnal cycle in normothermic and torpid African blue-naped mousebirds (Urocolius macrourus). AB - Colies are one of the phylogenetically oldest groups among the modern birds; the earliest finds are from about 35 million years ago. In states of energy deficiency they can undergo torpor during the night when metabolic rate and body temperature are decreased drastically to save energy (up to 90%). Here, we report the first measurements of heart rate (HR) by long-term telemetry, in seven individuals of blue-naped mousebirds (Urocolius macrourus); simultaneously and continuously metabolic rate (MR) was determined. HR at night was about 20% below the range of expected values (246/310 bpm). Mean oxygen pulse (O2 output/stroke) in normothermic birds was in a range of 0.019-0.020 ml O2/stroke; during torpor nights this value decreased significantly to 0.0086. Mean cardiac output ranged from 724 to 1214 ml blood/kg per min; in torpid birds this value fell to 400 ml blood/kg per min. Cardiac regulation of metabolic demand within an activity phase (day or night) is mainly achieved by chronotropy. Inotropy contributes at most 25% to the differences in MR between day and night (ca. 40%). Entry into torpor is brought about mainly by changes in HR (decrease from 240 to 90 bpm); after torpor levels have been reached, there is an increase in HR (to 200 bpm) and a sharp decrease (-53%) in stroke volume. This regulation by inotropy is also characteristic of arousal from torpor. PMID- 10682243 TI - Understanding autonomic sympathovagal balance from short-term heart rate variations. Are we analyzing noise? AB - Heart rate variations reflect the output of the complex control of the heart mediated by the autonomic nervous system. Because of that, they also encode different types of information, namely the efferent outflow of reflex mechanisms involved in the beat-to-beat control of cardiac function, the efferent activity of neurohumoral elements involved in the control of other cardiovascular parameters and random noise resulting from the hysteresis of the different controllers. The degree to which power spectrum estimation methods will uncover the periodic component of heart rate variations is in direct relation with the status of the system under study. Although the utility of spectral methods is now established in mammalian research, very little is known on the utility of these techniques in non-mammalian cardiovascular research. This review covers this space by discussing the physiological significance of heart rate variations in non-mammalian vertebrates. A detailed account of the different steps of the technique, its limitations and the ways to overcome these problems are also presented. These are: the recording of the cardiac event signal, the detection and digital processing methods, the satisfaction of stationarity conditions, the problem of spectral leakage and the different methods to estimate the power spectrum. PMID- 10682244 TI - Cardiac rhythms in developing chicks. AB - Instantaneous heart rate (IHR) of chicks was determined by electrocardiogram measured non-invasively from the day of hatch to day 6 for continuity of investigation of HR fluctuations from embryos and for ascertainment of HR diurnal rhythms. In Experiment I, IHR was determined for 1-h periods twice a day, in daytime and at night, to investigate development of heart rate fluctuations (variability and irregularities). Chick IHR was substantially more arrhythmic than embryonic HR and spontaneous acceleration dominated HR fluctuations. Chick HR fluctuations were categorized into three types; [1] Type I as a widespread baseline HR (20-50 bpm) due to respiratory arrhythmia, with a mean oscillatory frequency of 0.74 Hz (range 0.4-1.2 Hz); [2] Type II as low frequency oscillations of baseline HR, at a mean of 0.07 Hz (range 0.04-0.10 Hz), and [3] Type III as non-cyclic irregularities, dominated by frequent transient accelerations. In Experiment II, continuous measurements of HR were made under conditions of a natural photoperiod, thermoneutrality and with feed available throughout the first week after hatching and circadian rhythms of HR were ascertained. HR was very variable in the daytime (250-500 bpm), due in part to feeding and activity, and decreased to a diurnal low (200-350 bpm) at night when mean HR was relatively stable. HR fluctuations persisted throughout the diurnal cycle. PMID- 10682245 TI - Dynamical systems analysis of arterial blood pressure signals in relation to heart rate fluctuations in chick embryos. AB - We attempted a new approach based on a modern dynamical system theory to reconstruct the arterial blood pressure signals in relation to heart rate fluctuations of developing chick embryos. The dynamical systems approach in general is to model a phenomenon that is presented by a single time series record and approximate the dynamical property (e.g. heart rate fluctuations) of a system based only on information contained in a single-variable (arterial blood pressure) of the system. The time-series data of the arterial blood pressure was reconstructed in 3-dimensional space to draw characteristic orbits. Since the reconstructed orbits of the blood pressure should retain information contained in the pressure signals, we attempted to derive instantaneous heart rate (IHR) from the reconstructed orbits. The derived IHR presenting HR fluctuations coincided well with the IHR obtained conventionally from the peak-to-peak time intervals of the maximum blood pressure. Movements of the reconstructed orbits of the arterial blood pressure in 3-dimensional space reflected HR fluctuations (i.e. transient decelerations and accelerations). PMID- 10682246 TI - Development of cardiac rhythms in altricial avian embryos. AB - Mean heart rate (MHR) was determined during incubation and in hatchlings of 14 altricial avian species to investigate (1) if there is a common developmental pattern of heart rate in altricial embryos and (2) if heart rate changes during incubation are correlated with changes in embryonic growth rate. On the basis of normalized incubation MHR increased approximately linearly in 12 of 14 species from as early as 30-40% of incubation to that of pipped embryos. The MHR of hatchlings was equal to or higher than that of pipped embryos in seven species. Passerine embryos and hatchlings maintained higher MHR in comparison to parrots of similar egg mass, which may reflect phylogenetic differences in development. Embryonic MHR increased at a higher rate while embryonic growth rates were highest during the first 40% of incubation in tit, budgerigar and crow embryos than during subsequent development when relative growth rates decreased. MHR became independent of yolk-free wet mass at a smaller fraction of hatchling mass in budgerigar and crow than in the tit, suggesting that MHR is more likely to increase continuously after 40% of incubation in small altricial species than larger species. PMID- 10682247 TI - Long-term measurement of heart rate in chicken eggs. AB - Taking advantage of acoustocardiogram (ACG), we measured the heart rate (HR) of chick embryos continuously from day 12 until hatching and then investigated the development of HR irregularities (HRI), HR variability (HRV), and the existence of a circadian rhythm in mean HR (MHR). HRI comprised transient bradycardia and tachycardia, which first developed on day 14 and 16 in most embryos, respectively. Transient bradycardia increased in frequency and magnitude with embryonic development and occurred over periods of up to 30 min in some embryos. MHR was maximal on around days 14-15 and thereafter decreased to about 250-260 bpm on days 16-18. Baseline HRV, which is an oscillation of the MHR baseline, occurred as HR decreased from days 15-16 and became predominant on days 17-18. The magnitude of the baseline oscillations reached up to 50 bpm in some embryos and the period ranged between about 40-90 min (ultradian rhythm). A circadian rhythm of MHR was not found in late chick embryos. On days 18-19, embryonic activities were augmented and then breathing movements began to occur, disturbing ACG signals and thus making it difficult to measure the HR. Instead, the development of breathing activities was recorded. Breathing frequency was irregular at first and then increased to a maximum of about 1.5 Hz prior to hatching. PMID- 10682248 TI - Analysis of heart rate in developing bird embryos: effects of developmental mode and mass. AB - Bird embryos may be regarded as developing in their thermo-neutral zone, at rest, and stay in the egg for a fixed period of time until hatching. It is therefore interesting to investigate if they follow the same 'rule' set for adult homeotherms, which states that, within a taxonomically or functionally defined category such as mammals or birds, the number of heart beats throughout the life span (sL) is more or less constant. This rule stems from the allometric relationships between heart rate (fH) and body mass (mB) and between sL and mB. As a step towards understanding the general allometric nature of avian embryonic physiology we analyzed the fH values of avian embryos in relation to their incubation span (sI). Data from 30 species were selected from the scientific literature for the analyses. Values obtained from invasive methods which were judged to grossly alter natural incubation conditions, or from undefined or unmatched temperature conditions were not used. These include most values obtained below the first 30% of the incubation. Also, data obtained after internal pipping were discarded since hatching activity influences them. Values for sI and egg mass (mE) as representatives of embryonic mass were also collected. Embryonic fH was normalized to 70.1-80% sI. At 20.1-30% sI it was only 85% of the value at 70.1-80% sI and increased to a plateau at about 50.1-60% sI. It was almost constant among species between 50.1 and 60% sI and pre-internal pipping (PIP) time and thus, the mean fH value between 50.1 and 60% sI and between 90.1 and 100% excluding pipped eggs (fH) was taken as a representative value for each given species. The fH (min-1) and the corresponding sI (days) values for the 30 species, scaled with mE (g) as follows: fH = 371.1.mE-0.112 and: sI = 12.29.mE+0.209. Both powers were significantly different from 0. The product of fH and sI (fH.sI), representing the total number of heartbeats throughout the incubation, scaled with mE for the entire data set as follows: fH.sI = 6.565 x 10(+6).mE+0.096, where the +0.096 power is significantly different from 0. Values for fH.sI from embryos of altricial birds tended to concentrate at the low mE end of the plot while those of the precocial ones tended towards the high end. Separate analyses showed that the mE power for the combined altricial and semi-altricial species (ASA), and the combined precocial and semi precocial species (PSP), of log fH.sI against log mE regressions, were both insignificantly different from 0. Thus, means of fH.sI for ASA and PSP were calculated. The mean ASA value of 7.27 x 10(+6) heartbeats for fH.sI, was significantly different from the mean PSP value of 10.93 x 10(+6). The difference of 3.66 x 10(-6) (33.5%) heartbeats can be attributed to either the more advanced stage of the PSP hatchlings at hatch, to the larger mE values of these hatchlings, to the difference in water fraction of the hatchlings or all. The result of a linear regression of fH.sI against the rate of sI completion (the inverse of incubation span, fI; day-1) was: fH.10(-6) = 0.205 + 3.940.sI-1. Thus, the faster is the average rate of development accomplished per day (shorter incubation) the higher is daily heart rate. Data tended to cluster such that large eggs, mostly of the PSP type with relatively low fH, complete 2-4% of their incubation per day, while small, ASA type eggs with relatively high fH, complete 6-8% of their incubation time per day. We conclude that, at this stage of knowledge, the data is insufficient to resolve whether the different modes of hatch stage alone can explain differences in the total number of heartbeats throughout embryonic life among all bird species, or egg mass and water content differences contribute variability. This should be investigated on a larger sample of species in more depth. PMID- 10682249 TI - Development of heart rate rhythmicity in Muscovy duck embryos. AB - The heart rate (HR) of Muscovy duck embryos (Cairina moschata f. domestica) was continuously recorded from as early as the 21st day of incubation (D21) until hatching (D34/35). The aim of the study was to investigate the influence of phonoperiods consisting of different acoustic stimuli on the course of HR and the development of HR periodicities during this period. Incubation was carried out at a constant temperature and in constant darkness. Until D25 HR was dominated by decelerative fluctuations only, indicating a main input from the parasympathetic system on the heart. Later sympathetic influences increased progressively. HR periodicity was investigated by means of chi 2-periodogram and fast Fourier transformation. Between D26 and D30 statistically significant and stable HR periodicities developed gradually. They had periods in the range from 5 to 38 h. Ultra-, circa- and infradian rhythms (< 20, 24 +/- 4 and > 28 h, respectively) occurred in parallel in some cases in the same embryo. The for the HR course important periods were dissimilar between individual embryos and had different intensities. There was no indication that acoustic stimulation (phonoperiods) had any effect on the development of HR periodicities. PMID- 10682250 TI - Cardiac rhythms in chick embryos during hatching. AB - Avian embryos develop within a hard eggshell which permits the measurement of heart rate while maintaining an adequate gas exchange through the chorioallantoic membrane. Heart rate has been determined from cardiogenic signals detected either noninvasively, semi-invasively or invasively with various transducers. Firstly, we reviewed these previously-developed methods and experimental results on heart rate fluctuations in prenatal embryos. Secondly, we presented new findings on the development of heart rate fluctuations during the last stages of incubation, with emphasis on the perinatal period, which remained to be studied. Three patterns of acceleration of the instantaneous heart rate were unique to the external pipping period: irregular intermittent large accelerations, short-term repeated large accelerations and relatively long-lasting cyclic small accelerations. Besides these acceleration patterns, respiratory arrhythmia, which comprimised oscillating patterns with a period of 1-1.5 s, appeared during the external pipping period. Furthermore, additional oscillating patterns with a period of 10 15 min were found in some externally pipped embryos. PMID- 10682251 TI - Cardiac responses to first ever submergence in double-crested cormorant chicks (Phalacrocorax auritus). AB - Heart rates were recorded from double-crested cormorant chicks during their first ever and subsequent voluntary head submergences and dives, as well as during longer dives made after the chicks were accustomed to diving. Despite variation between chicks, the cardiac response to first ever and subsequent voluntary submergence (head submergences and dives) was similar to the response observed in adult cormorants. Upon submersion the heart rate fell rapidly when pre-submersion heart rate was high (325-350 beats min-1). The heart rate established within the first second of voluntary submergence was between 230 and 285 beats min-1, well above resting heart rate (143 beats min-1). The same initial cardiac response occurred during longer dives performed after the chicks were accustomed to diving. In these dives the heart rate remained at the level established on submersion, unlike the response observed in shallow diving adult cormorants in which the heart rate declined throughout the dive. The heart rate was also monitored in a separate group of chicks in which the first exposure to water was during whole body forced submergence. Again, the observed response was similar to the adult response, although the cardiac response of chicks to forced submergence was more extreme than to voluntary submergence. Our results do not support the hypothesis that learning (by conditioning or habituation) is involved in the cardiac adjustments to voluntary submergence. It is suggested that the initial cardiac adjustments are reflex in nature and this reflex is fully developed by the first submergence event. Although the nature of this reflex pathway is obscure, cessation of breathing before submersion and the close linkage between breathing and heart rate might provide a plausible mechanism. PMID- 10682252 TI - MR and CT angiography of the pelvis: clinical and technical considerations. PMID- 10682253 TI - Radiographic equipment, installation, and radiation protection. PMID- 10682254 TI - Spatial analytical methods and geographic information systems: use in health research and epidemiology. PMID- 10682255 TI - Hepatitis E: an emerging infectious disease. PMID- 10682256 TI - Hepatitis C-related hepatocellular carcinoma: prevalence around the world, factors interacting, and role of genotypes. PMID- 10682257 TI - Epidemiologic studies of leukemia among persons under 25 years of age living near nuclear sites. PMID- 10682258 TI - Epidemiology of abdominal aortic aneurysms. PMID- 10682259 TI - Role of cannabis in motor vehicle crashes. PMID- 10682260 TI - Analytical studies of enamel fluorosis: methodological considerations. PMID- 10682261 TI - Longitudinal change in the heights of men and women: consequential effects on body mass index. PMID- 10682262 TI - Epidemiology of gestational weight gain and body weight changes after pregnancy. PMID- 10682263 TI - The unexpected promiscuity of steroid hormones. PMID- 10682264 TI - Synaptonemal complex analysis in human male infertility. AB - The fine structural features of human spermatocytes from carriers of some of the most frequent chromosomal abnormalities are reviewed on the basis of original data and previous reports from the literature. Special emphasis is given to the Robert-sonian translocations t (13; 14), to one specific reciprocal translocation involving chromosome 21, and to Y disomy in spermatocytes from XYY men. Synaptonemal complex analysis shows that in many carriers of chromosomal aberrations that lead to pachytene configurations having terminal asynaptic segments in autosomes, there is a gradual association of these asynaptic segments with the XY body. This associations with the XY pair is assumed to trigger a process of germ cell deterioration, presumably through the spreading of the X chromosome inactivation towards autosomal segments. Another different process of germ cell deterioration occurs when the X chromosome becomes an univalent, as in XYY men with persistence of two Y chromosomes in the germ line. The renewed interest in the examination of spermatocytes from human testicular biopsies is commented upon. PMID- 10682265 TI - Presence and localization of molecules related to the cholinergic system in developing rat testis. AB - The presence of cholinergic molecules was recently found in male gametes of different animal species; very little is known from this point of view about the somatic component of the gonad. In the present study, a contribution is given to the localization of some cholinergic-like molecules, such as acetylcholinesterase (AChE), nicotinic acetylcholine and muscarinic acetylcoline (nAchR and mAChR, respectively) receptors during postnatal development of the testis. Maturation stages were checked by use of histochemical stainings, such as DAPI for nuclear shape changes, and PSA binding to reveal acrosomal maturation. The distribution of cholinergic-like molecules, revealed by immunocytochemical methods in both gametes and somatic cells, varied with development. Generally, during early stages, molecules immunologically related to AChE, and to mAChR's were mainly found in the interstitial and peritubular compartment, while, during maturation, they were found in Sertoli cells and in differentiating germ cells. nAChR's were not found in the somatic compartment, except in the blood vessel walls, and were distributed in mature germ cells, mainly in the flagella. The presence of cholinergic molecules in somatic as well as germ cells could play a role in cell to-cell communications affecting testicular cell differentiation and function. PMID- 10682266 TI - Tubulin isoforms are differently expressed in developing and mature neurons: a study on the cerebral cortex of newborn and adult rats. AB - Tubulin heterogeneity was observed in the rat brain, where these proteins can vary in different neurons suggesting multiple functions. In this paper, the different expression of tubulins in cerebral cortex between newborn and adult rats was analyzed by Western blot and immunocytochemical methods, using anti tubulin antibodies. Our results showed that tubulins were present at higher levels in the newborn than in the adult cerebral cortex. In newborn rats, a marked staining of the perikarya and basal dendrites of pyramidal cells was noted. This significant expression of tubulins in the newborn cerebral cortex could be related to the major needs of tubulins in developing neurons. The higher amount in tyrosine-tubulin and class III beta-tubulin could be consistent with the state of "dynamic instability", typical of the microtubular network of neurons during brain development. PMID- 10682267 TI - Occurrence of GABA-transaminase in the thymus gland of juvenile and aged rats. AB - The occurrence and distribution of GABA-transaminase (GABA-t) activity were examined in the thymus of juvenile, adult and aged rats, using enzyme histochemical and biochemical methods. Quantitative image analysis showed that specific GABA-t reactivity was localized in the wall of the arteries and, to a lesser extent, to the veins. Only a low activity could be observed in association with the subcapsular and medullary part of the parenchyma of the thymus. Many structures resembling nerve fibers also showed low positive reactivity. Biochemical results gave the following decreasing order of GABA-t activity: arteries, veins, whole thymus and parenchyma. Histoenzymatic staining and related values of quantitative analysis of images, are in agreement with the biochemical results; moreover, they demonstrated that the intensity of histoenzymatic staining for GABA-t in thymus of rats strongly decreases with age. GABA-t in thymic tissue is concentrated in blood vessels and particularly in the arteries. Therefore, our findings do not support the earlier assumptions that GABA-t is exclusively concentrated in cerebral vessels. Moreover, the decrease of GABA-t activity during age in thymic tissues may be related to the decrease of thymic microvessels as consequence of the thymic involution. On the contrary, the thymic macrovessels show an elevated staining in all ages with an apparent increase due to the thymic involution. PMID- 10682268 TI - Developmental pattern of NADPH-diaphorase activity in the peripheral nervous system of the cichlid fish Tilapia mariae. AB - The distribution of NADPH-diaphorase activity was studied in the cichlid fish Tilapia mariae, during the first developmental stages by means of the tetrazolium salt technique. The reaction product was first found, 48 hours after fertilization (stage 10), in the cells of the olfactory placodes and in the superficial neuromasts. A faint positivity was seen in some hair cells of the otic vesicles. The epithelial cells of the most caudal part of the intestinal tract showed a strong labeling. At stage 12 (hatch), the reaction product was in addition detected in scattered enteric neurons surrounding the digestive tract. At stage 13 (4.5 days after spawning), the reaction product was also found in the putative sympathetic trunk, which supplies the gill arches and digestive tract. The epithelial cells of the gastrointestinal canal showed a more strong positive labeling and two large clusters of cells near the pronephritic tubules (the putative adrenomedullar tissue) were also labeled. The present results indicate an early activity of NADPH-diaphorase during the development of the peripheral nervous system of Tilapia and reveal a gradual maturation of NADPH-diaphorase positive structures. PMID- 10682269 TI - Immunohistochemical detection of mutant p53 protein in regional lymph nodes is associated with adverse outcome in stage II colorectal cancer. AB - Epidemiologic data identifies a cohort of Duke's B (CRC) patients whose survival more closely matches that of Duke's C. Lymph node micrometastases may account for this discrepancy. Lymph node expression of mutant p53 protein (Mp53P) has been linked to a reduction in survival in Japanese Duke's B patients. We aimed to determine the significance of nodal p53 expression in European Duke's B patients using immunohistochemistry. The study comprised 134 consecutive patients who had resections for CRC between 1984 and 1991. End points were 5 year disease free survival or CRC related death. Thirty-four subjects did not achieve end points and were excluded. We examined tumour and nodal sections for Mp53P by immunohistochemistry and correlated this with survival using a Kaplan-Meier (KM) and a Cox Proportional hazards model (CPHT). Five year survival was 73%. Fifty eight percent of primary tumours expressed Mp53P. Tumour p53 expression did modulate survival behavior. Twenty-six percent of subjects' lymph nodes expressed Mp53P. Fifty-three per cent of those with positive and 17% of those with negative lymph nodes died of recurrence. The relative risk for nodal Mp53P expression was 3.1. There was a significant univariate relationship between lymph node p53 expression and mortality. (Log Rank p = 0.028). Multivariate analysis also showed a significant relationship with mortality. (CPHT p = .03). We conclude that lymph node expression of Mp53P is associated with increased mortality in Duke's B CRC. PMID- 10682270 TI - Immunohistochemical detection of lactoferrin in human astrocytomas and multiforme glioblastomas. AB - The presence of lactoferrin in astrocytomas, anaplastic astrocytomas and multiforme glioblastomas was determined by immunohistochemistry; the staining intensity and the percentage of neoplastic stained cells were graded and statistical analysis was performed by non-parametric methods. A moderate to strong diffuse immunoreactivity for lactoferrin was shown in glial elements of astrocytomas, while the positivity was progressively reduced in anaplastic astrocytomas and in multiforme glioblastomas, some of which were unstained; a highly significant difference was found between scores relative to astrocytomas and glioblastomas. We suggest that the lactoferrin may be produced by neoplastic astrocytes which permits a greater availability of iron for metabolic cellular processes. Alternatively, the cytoplasmic localization of lactoferrin in neoplastic astrocytes may be the consequence of defective or functionally impaired lactoferrin receptors at the cellular surface. PMID- 10682271 TI - Autometallographical localisation of Cu and Zn within target cell compartments of winkles following exposure to Cu&Zn mixtures. AB - This investigation attempts to determine the usefulness of autometallography to localise particular metals in certain key tissues of molluscs exposed to metal mixtures. For this purpose, winkles (Littorina littorea) removed from shell were exposed to very high concentrations of either copper (Cu), zinc (Zn) or a mixture of both metals (Cu&Zn) dissolved in sea-water for short periods of time. Protein bound metals were detected by autometallography as black silver deposits (BSD) on histological sections of gills, foot, mantle, digestive gland/gonad complex, stomach and kidney. Copper was localised within cytoplasmic granules of gill ciliated cells, nephrocytes and stomach epithelial cells as well as within digestive cell lysosomes. Zinc was essentially found in the basal lamina (histological sense) of gill, stomach, kidney and digestive gland epithelia. BSD were also evidenced in cytoplasmic granules of pore cells present in parenchymal connective tissue of mantle, foot, gill, digestive gland and stomach. Copper and zinc concentrations were additionally calculated for the whole soft body as well as for certain organs by atomic absorption spectrophotometry (AAS). According to AAS, a synergistic phenomenon would contribute to increase the rate of Cu and Zn accumulation in presence of each other. However, after exposure to Cu&Zn autometallography did not evidence any synergistic phenomenon, and Cu and Zn were localised in their respective accumulation sites. In conclusion, autometallography might indicate the presence of certain metals in the environment irrespective of factors, such as "metal-metal interaction-like" phenomena, affecting metal concentrations in soft tissues. PMID- 10682272 TI - Immunoreactivity of the monoclonal antibody F89/160.1.5 for the human prion protein. AB - Monoclonal antibodies (Mab) to the prion protein (PrP) have been critical to the neuropathological characterisation of PrP-related diseases in human and animals. Although PrP is highly evolutionary conserved, there is some sequence divergence among species. We have analysed the F89/160.1.5 Mab raised against the bovine prion protein for immunoreactivity with the human prion protein. The antibody recognised the IHFG epitope of the prion protein. An analysis of the Swiss Prot database confirmed conservation of the epitope in humans. Further immunohistochemical (IHC) analysis showed a highly sensitive (final concentration 55 ng/ml) and specific antibody for prion detection in humans. The observed immunoreactivity of the prion protein did not differ from that observed after staining with the well-known 3F4 (Senetek) monoclonal antibody. PMID- 10682273 TI - Confocal laser microscopy to investigate myoepithelial cells in tissue blocks. PMID- 10682274 TI - The estimation of marginal time preference in a UK-wide sample (TEMPUS) project. PMID- 10682275 TI - Evaluation of oxygen permeability of gas vesicles from cyanobacterium Anabaena flos-aquae. AB - The enhancement of oxygen permeability in aqueous medium by addition of cyanobacterial gas vesicles (GVs) has been examined. The GVs were isolated from cultures of Anabaena flos-aquae that had been cultivated in photobioreactors and harvested by dark flotation. Prior to the permeability experiments, the collected GVs were treated with glutaraldehyde for improved stability. Measurements of oxygen permeability were made with a polarographic oxygen electrode in suspensions of various GV volume fractions (0-2.1%). The experimental results were compared with the values predicted theoretically (Fricke's equation) assuming different permeability through the GVs (PmGV), ranging from 0 to 8.30 x 10(-4) mol m-1 atm-1 s-1. The former corresponded to impermeable vesicles, the latter to air at 22 degrees C as if there were no vesicle wall. The best-fit value of PmGV was 9.9 x 10(-7) mol m-1 atm-1 s-1, ca. 36-fold higher than that in water. GVs were therefore very permeable to oxygen. However, the value was much lower than that predicted for air, implying the existence of wall resistance. PMID- 10682276 TI - Large-scale production of recombinant hepatitis B surface antigen from Pichia pastoris. AB - The ability of the Pichia pastoris-based technology for large-scale production of recombinant hepatitis B virus surface antigen (HBsAg) and both reproducibly purify HBsAg and remove most of the relevant contaminants was ascertained by evaluating ten industrial production batches, five in 1993 and five in 1998. At an early stage, the clarification of mechanically disrupted yeast cells by acid precipitation renders HBsAg with a purity as low as 3.8 +/- 0.6%. However, by adsorption/desorption from diatomaceous earth matrix, the purity of HBsAg rapidly increases to 18.8 +/- 5%, which is suitable for chromatographic processing. This step also eliminates non-particulated forms of HBsAg, significantly lowers the amount of carbohydrates and lipids, and concentrates the HBsAg 4.8-fold. Finally, a sequential purification procedure that includes large-scale immunoaffinity, ion exchange, and size-exclusion chromatographies further purifies the preparation, resulting in a product (HBsAg at a concentration of 1.3 +/- 0.2 g l-1) with a purity of 95% or more. Furthermore, each of the other contaminants measured reaches the following low levels per 20 micrograms HBsAg: host deoxyribonucleic acid (< 10 pg), carbohydrates (1.2 +/- 0.02 micrograms), lipids (14 +/- 0.28 micrograms), immunopurification-released immunoglobulin G (less than 100 ppm), and endotoxins (106.7 +/- 19.3 pg). These values are below those specified for recombinant DNA hepatitis B vaccines according to World Health Organization (WHO) guidelines. PMID- 10682277 TI - Production and in vitro refolding of a single-chain antibody specific for human plasma apolipoprotein A-I. AB - An active form of single-chain antibody (scFv) has been produced in Escherichia coli for murine monoclonal antibody MabA34 (gamma 1, kappa), which is specific for human plasma apolipoprotein (apo) A-I. The complementary DNAs (cDNAs) encoding the variable regions of heavy chain (VH) and light chain (VL) were connected by a (Gly4Ser)3 linker using an assembly polymerase chain reaction. The construct (VL-linker-VH) was placed under the control of highly efficient T7 promoter system. The cloned scFv was expressed in E. coli as inclusion bodies. After purification from E. coli lysate using sonication and low speed centrifugation, the inclusion body was solubilized and denatured in the presence of 8 M urea, renatured by dialysis, and scFv was finally purified using antigen affinity chromatography. The purity and activity of purified scFv were confirmed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), Western blotting and enzyme-linked immunosorbent assay (ELISA). The affinity constant was determined by a biosensor method using the BIAcore system. The results showed that the yield of correctly refolded scFv was more than 20 mg l-1 of E. coli flask culture and the specific binding activity to apo A-I was retained with an affinity constant of 6.74 x 10(-8) M (Kd). A notable thing is that guanidine-HCl as a denaturant induced more multimeric formation in the subsequent refolding procedure for the scFv of MabA34 and thus, it was not suitable as urea was. This fact is uncommon for what is generally known for the denaturation and refolding of recombinant antibodies. PMID- 10682278 TI - The stimulation of gene expression by the R region from HTLV-1 and BLV. AB - The 5' untranslated regions (5'UTR) of mRNA are known to stimulate or inhibit more or less translation. SR alpha, an association of SV40 early gene promoter and of the R region plus the first 39 nucleotides of the U5 region (designated as R) from the human T-cell leukemia virus (HTLV-1) is currently used to stimulate expression of various coding regions. Its effect is considered to take place at the translational level. In all studies published so far, the R region was associated with the promoter and 5'UTR from SV40 early genes. In the present work, the role of SV40 5'UTR and HTLV-1R region was evaluated separately using different promoters, reporter genes and cells. Both SV40 5'UTR (SU) and R region (R) from HTLV-1 stimulated separately the expression of adjacent reporter genes. When associated, the SV40 5'UTR and the R region from HTLV-1 (SUR) were a more potent stimulator of gene expression and their effects were more than additive. This effect was very potent in HeLa and HC11 cells and almost inexistent in CHO and COS 7 cells. It was of various intensity in other cell types including bird and fish cells. The presence of SUR in gene constructs favoured the accumulation of the mRNAs. SUR stimulated gene expression when added between the cap and the initiation codon. Unexpectedly, SUR was never inhibitory. SUR can therefore be considered essentially as potent and specific stimulator of gene expression favoring mRNA accumulation. PMID- 10682279 TI - High production of 1,3-propanediol from industrial glycerol by a newly isolated Clostridium butyricum strain. AB - Batch and continuous cultures of a newly isolated Clostridium butyricum strain were carried out on industrial glycerol, the major by-product of the bio-diesel production process. For both types of cultures, the conversion yield obtained was around 0.55 g of 1,3-propanediol formed per 1 g of glycerol consumed whereas the highest 1,3-propanediol concentration, achieved during the single-stage continuous cultures was 35-48 g l-1. Moreover, the strain presented a strong tolerance at the inhibitory effect of the 1,3-propanediol, even at high concentrations of this substance at the chemostat (e.g. 80 g l-1). 1,3 Propanediol was associated with cell growth whereas acetate and butyrate seemed non growth-associated products. At low and medium dilution rates (until 0.1 h-1), butyrate production was favoured, whereas at higher rates acetate production increased. The maximum 1,3-propanediol volumetric productivity obtained was 5.5 g l-1 h-1. A two-stage continuous fermentation was also carried out. The first stage presented high 1,3-propanediol volumetric productivity, whereas the second stage (with a lower dilution rate) served to further increase the final product concentration. High 1,3-propanediol concentrations were achieved (41-46 g l-1), with a maximum volumetric productivity of 3.4 g l-1 h-1. A cell concentration decrease was reported between the second and the first fermentor. PMID- 10682280 TI - Intracellular glucose 6-phosphate content in Streptomyces coelicolor upon environmental changes in a defined medium. AB - A new, chemically defined medium providing dispersed growth and high biomass formation and a method for quantitative extraction of intracellular metabolites was used to investigate the cellular response of Streptomyces coelicolor A3(2) during growth and upon changes in nutrient utilization. Fast changes of the glucose 6-phosphate content precisely signaled transitions in the flow of carbon sources. The results indicate that intracellular pool sizes may be used to detect early nutrient limitations in view of the onset of antibiotic production. Additionally the results disclose characteristics of the regulation of maltose and glutamic acid uptake and degradation in S. coelicolor A3(2). PMID- 10682281 TI - Engineering of a proteolytically stable human beta 2-adrenergic receptor/maltose binding protein fusion and production of the chimeric protein in Escherichia coli and baculovirus-infected insect cells. AB - The hydrophobic human beta 2 adrenergic receptor was produced in fusion to the hydrophilic maltose-binding protein (MalE) in Escherichia coli. Photoaffinity labeling with the adrenergic ligand [125I]cyanopindolole-diazirine indicated that the majority of the protein was proteolyzed in the intergenic region between the fusion partners after production in E. coli. The simple and fast genetics of the bacterium enabled us to engineer a linker with an increased proteolytic stability. The fusion protein produced in E. coli was fully functional with respect to binding of adrenergic ligands and coupling to stimulatory GTP-binding protein. The production level with 3 pmol receptor fusion protein per mg membrane protein in a crude membrane preparation was significantly higher than those reported for other beta 2 adrenergic receptor constructs in E. coli. After solubilization with dodecanoyl sucrose, the fusion protein was purified to near homogeneity by affinity chromatography on immobilized Ni2+ ions (binding to a C terminal His6-tag) and on crosslinked amylose (binding to the MalE). In order to achieve higher production levels, the fusion protein preceded by an insect signal peptide was produced in baculovirus-infected insect cells. As expected, the production level with about 17 pmol receptor per mg membrane protein was higher in the insect cells than in E. coli. The receptor fusion protein produced in the insect cells bound adrenergic ligands and activated heterotrimeric GTP-binding proteins with biochemical properties comparable to that of the unfused receptor. PMID- 10682282 TI - Different fungal manganese-oxidizing peroxidases: a comparison between Bjerkandera sp. and Phanerochaete chrysosporium. AB - Two manganese-oxidizing peroxidases differing in glycosylation degree were purified from fermenter cultures of Bjerkandera sp. They were characterized and compared with the three manganese-oxidizing peroxidase isoenzymes obtained from the well-known ligninolytic fungus Phanerochaete chrysosporium. All the enzymes showed similar molecular masses but those from P. chrysosporium had less acidic isoelectric point. Moreover, the latter strictly required Mn2+ to oxidize phenolic substrates whereas the Bjerkandera peroxidases had both Mn-mediated and Mn-independent activity on phenolic and non-phenolic aromatic substrates. Taking into account these results, and those reported for Bjerkandera adusta and different Pleurotus species, we concluded that two different types of Mn(2+) oxidizing peroxidases are secreted by ligninolytic fungi. PMID- 10682283 TI - Calorimetrically recognized maximum yield of poly-3-hydroxybutyrate (PHB) continuously synthesized from toxic substrates. AB - The broader usage of poly-beta-hydroxybutyrate (PHB), for instance as bulk plastics, calls for cheap raw materials and greater overall process efficiency. The bacterial synthesis is generally induced and promoted by the limitation of growth via nitrogen, oxygen or phosphate depletion with the simultaneous excess and higher concentration of the carbon substrate. Consequently, toxic substrates have been considered unsuitable for PHB synthesis. Nevertheless, a single-stage continuous process for producing PHB from toxic substrates using microorganisms was developed and is reported here. The maximum heat flux during continuous growth and the maximum yield of PHB versus the substrate consumption rate were found to coincide. This suggests the possibility of controlling the conversion of a growth-inhibiting substrate into PHB and maximizing the process efficiency. The observed correlation occurred irrespective of the substrates investigated (phenol or sodium benzoate), the PHB-producing strain (Ralstonia eutropha JMP 134 or Variovorax paradoxus JMP 116), or the type of limitation imposed. The maximum PHB yields obtained comprised up to 50% of cell dry mass. PMID- 10682284 TI - Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2 alpha-D-mannosidase. AB - A cDNA encoding 1,2-alpha-D-mannosidase mds 1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56,266 Da and shows high similarity to the amino acid sequences of 1,2-alpha-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0% identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase. PMID- 10682285 TI - Oxirane-immobilized Lentinula edodes laccase: stability and phenolics removal efficiency in olive mill wastewater. AB - Immobilization of Lentinula edodes laccase on Eupergit C increased pH, thermal and proteolytic stability with slight modifications in laccase oxidation efficiency. Immobilized laccase proved to be efficiently stable in removing olive mill wastewater phenolics. PMID- 10682286 TI - Hydrolysis of oleuropein by recombinant beta-glycosidase from hyperthermophilic archaeon Sulfolobus solfataricus immobilised on chitosan matrix. AB - The recombinant beta-glycosidase (EcS beta gly) from Sulfolobus solfataricus was immobilised on chitosan to perform the enzymatic hydrolysis of commercial oleuropein (heterosidic ester of elenolic acid and 3,4-dihydroxy-phenylethanol (hydroxytyrosol)) at two temperatures (60 and 70 degrees C). Interestingly, on the basis of the reasonable assumption that the enzyme hydrolyses only the sugar linkage, the biotransformation produces unstable aglycone species formed by oleuropein hydrolysis that, differently from some commercially available beta glucosidases tested, give rise to the formation of hydroxytyrosol, at the operative temperatures of the bioreactor. The results of the biotransformation at 70 degrees C showed that the main products are hydroxytyrosol, and glucose, being the oleuropein aglycone present in low amount at the end of reaction. Both in single step approach or in recycle approach the amounts of glucose and oleuropein aglycone were lightly dependent from flow rate. The amount of hydroxytyrosol, increased on decreasing the flow rate of bioreactor in recycle approach, following a non-linear trend and obtaining the highest value at a flow rate of 15 ml h-1 while in the single step approach the 3,4-dihydroxy-phenylethanol was at its maximum at higher flow rate (16 ml h-1). For the hydrolysis of the oleuropein by bioreactor at 60 degrees C we used lower molar ratio oleuropein/enzyme only by the single step approach. In these conditions it is possible to obtain high amounts of only two products (glucose and hydroxytyrosol) in short time (2 h). The stability of the bioreactor at the operative temperatures showed a t1/2 of 30 days at 70 degrees C and a t1/2 of 56 days at 60 degrees C. PMID- 10682287 TI - Epoxide hydrolase activity of Streptomyces strains. AB - The discovery of epoxide hydrolases within a Streptomyces sp. strain collection is described. Screening was performed in 96 well microtiter plates using a modified 4-(p-nitrobenzyl)pyridine assay with styrene oxide, 1,2-epoxy-hexane or 3-phenyl ethylglycidate (3-PEG) as substrates. Out of 120 strains investigated, S. antibioticus Tu4, S. arenae Tu495 and S. fradiae Tu27 exhibited epoxide hydrolase activity. These strains were further investigated by performing laboratory-scale biotransformations utilizing styrene oxide, 1,2-epoxy-hexane and 3-PEG followed by subsequent quantitative analysis employing chiral gas chromatography. The highest conversions were achieved with whole cells from S. antibioticus Tu4 in the presence of 10% (v/v) DMSO. However, enantioselectivity was only satisfying (E = 31) in the presence of 5% (v/v) acetone, which allowed isolation of optically pure non-hydrolyzed (R)-styrene oxide (99% enantiomeric excess (ee)) and (S)-phenyl-1,2-ethandiol (72% ee) at 55% conversion after 24 h. The resolution of 3-PEG proceeded with slightly lower enantioselectivity albeit higher reaction rates. With S. fradiae Tu27 and S. arenae Tu495 enantioselectivity towards styrene oxide was only E = 3-4. PMID- 10682288 TI - Thirty categorization results in search of a model. AB - One category structure dominated in the shift toward exemplar-based theories of categorization. Given the theoretical burden on this category structure, the authors reanalyzed 30 of its uses over 20 years in 8 articles. The authors suggest 4 conclusions. (a) This category structure may encourage exemplar memorization processes because of its poor structure, the learning difficulties it causes, and its small, memorizable exemplar sets. Its results may only generalize narrowly. (b) Exemplar models have an advantage in fitting these 30 data sets only because they reproduce a performance advantage for training items. Other models fit equally well if granted this capacity. (c) A simpler exemplar process than assumed by exemplar models suffices to explain these data sets. (d) An important qualitative result predicted by exemplar theory is not found overall and possibly should not even be expected. The authors conclude that the data produced by this category structure do not clearly support exemplar theory. PMID- 10682289 TI - Probability judgment in three-category classification learning. AB - People give subadditive probability judgments--in violation of probability theory -when asked to assess each in a set of 3 or more mutually exclusive hypotheses, as indicated by their sum exceeding 1. Three potential evidential influences on subadditivity--cue conflict, cue frequency, and cue redundancy--are distinguished and tested in 5 experiments using a classification-learning task. Results indicate that (a) judgments of probability and of frequency are systematically subadditive even when the judgments are based on cues learned within the experimental context, (b) cue conflict has a reliable influence on the degree of subadditivity, and (c) judgments in this context are well described by a linear discounting model within the framework of support theory. PMID- 10682290 TI - Competition among causes but not effects in predictive and diagnostic learning. AB - Causal asymmetry is one of the most fundamental features of the physical world: Causes produce effects, but not vice versa. This article is part of a debate between the view that, in principle, people are sensitive to causal directionality during learning (causal-model theory) and the view that learning primarily involves acquiring associations between cues and outcomes irrespective of their causal role (associative theories). Four experiments are presented that use asymmetries of cue competition to discriminate between these views. These experiments show that, contrary to associative accounts, cue competition interacts with causal status and that people are capable of differentiating between predictive and diagnostic inferences. Additional implications of causal model theory are elaborated and empirically tested against alternative accounts. The results uniformly favor causal-model theory. PMID- 10682291 TI - A feature-sampling account of the time course of old-new recognition judgments. AB - This article describes the feature-sampling theory of recognition (FESTHER), a new model of the time course of recognition judgments based on a model of the time course of perceptual processing in categorization (K. Lamberts, 1995, 1998). FESTHER is applied to previous results and to data from 4 old-new recognition experiments. Experiments 1 and 2 provided a preliminary test of the model's ability to explain recognition judgments of simple objects under response deadlines. Experiments 3 and 4 involved a response-signal procedure to elicit recognition judgments at different time lags after presentation of a stimulus. Simple objects and words were used as stimuli in Experiments 3 and 4, respectively. The new model accounts well for the data from the 4 experiments and offers a parsimonious account of the time course of recognition judgments based on the time-dependent availability of stimulus information. PMID- 10682292 TI - Automatic comparisons of artificial digits never compared: learning linear ordering relations. AB - In 2 experiments, participants were trained to perform magnitude decisions, that is, decide which of 2 arbitrary symbols in a pair represented a larger magnitude. The symbols corresponded to locations on an implicit linear scale. Training resulted in a Stroop-like size congruity effect when the participants had to decide which symbol in a pair was physically larger. This effect, showing automaticity of the processing of magnitude relations, was also obtained for pairs never encountered during practice. The implications of these findings for processing of magnitude relations and for theories of automaticity are discussed. PMID- 10682293 TI - Priming and attentional control of lexical and sublexical pathways during naming. AB - A modified priming task was used to investigate whether skilled readers are able to adjust the degree to which lexical and sublexical information contribute to naming. On each trial, participants named 5 low-frequency exception word primes or 5 nonword primes before a target. The low-frequency exception word primes should have produced a greater dependence on lexical information, whereas the nonword primes should have produced a greater dependence on sublexical information. Across 4 experiments, the effects of lexicality, regularity, frequency, and imageability were all modulated in predictable ways on the basis of the notion that the primes directed attention to specific processing pathways. It is argued that these results are consistent with an attentional control hypothesis. PMID- 10682294 TI - The role of physical and conceptual properties in preserving object continuity. AB - Six experiments investigated the nature of the object-file representation supporting object continuity. Participants viewed preview displays consisting of 2 stimuli (either line drawings or words) presented within square frames, followed by a target display consisting of a single stimulus (either a word or a picture) presented within 1 of the frames. The relationship between the target and preview stimuli was manipulated. The first 2 experiments found that participants responded more quickly when the target was identical to the preview stimulus in the same frame (object-specific priming). In Experiments 3, 4, 5, and 6, the physical form of the target stimulus (a word or picture in 1 frame) was changed completely from that of either preview stimulus (pictures or words in both frames). Despite this physical change, object-specific priming was observed. It is suggested that object files encode postcategorical information, rather than precise physical information. PMID- 10682295 TI - Updating displays after imagined object and viewer rotations. AB - Six experiments compared spatial updating of an array after imagined rotations of the array versus viewer. Participants responded faster and made fewer errors in viewer tasks than in array tasks while positioned outside (Experiment 1) or inside (Experiment 2) the array. An apparent array advantage for updating objects rather than locations was attributable to participants imagining translations of single objects rather than rotations of the array (Experiment 3). Superior viewer performance persisted when the array was reduced to 1 object (Experiment 4); however, an object with a familiar configuration improved object performance somewhat (Experiment 5). Object performance reached near-viewer levels when rotations included haptic information for the turning object. The researchers discuss these findings in terms of the relative differences in which the human cognitive system transforms the spatial reference frames corresponding to each imagined rotation. PMID- 10682296 TI - Path integration while ignoring irrelevant movement. AB - Participants attempted to return to the origin of travel after following an outbound path by locomotion on foot (Experiments 1-3) or in a virtual visual environment (Experiment 4). Critical conditions interrupted the outbound path with verbal distraction or irrelevant, to-be-ignored movements. Irrelevant movement, real or virtual, had greater effects than verbal or cognitive distraction, indicating inability to ignore displacement during path integration. Effects of the irrelevant movement's direction (backward vs. rightward) and location (1st vs. 2nd leg of path) indicated that participants encoded a configural representation of the pathway and then cognitively compensated for the movement, producing errors directly related to the demands of compensation. An encoding-error model fit to the data indicated that backward movement produced downward rescaling, whereas movement that led to implied rotation (rightward on 2nd leg) produced distortions of shape and scale. PMID- 10682297 TI - On the relation between representations constructed from text comprehension and transitive inference production. AB - Deductive inference production from texts is a process considered to involve either the construction of an integrated mental model or the step-by-step coordination of propositional representations of the sentences. These alternative hypotheses were tested in 3 experiments using a set inclusion task paradigm in which participants had to recall the premises and to evaluate transitive inferences. Contrary to what is known about linear ordering relations, order of recalls and reaction times provide evidence that the encoding of set inclusion relations does not result in an integrated representation. These results suggest that the mental models theory needs to take account of the nature of the relation to be represented if it is to become a general theory of reasoning. PMID- 10682298 TI - Metacognitive and control strategies in study-time allocation. AB - This article investigates how people's metacognitive judgments influence subsequent study-time-allocation strategies. The authors present a comprehensive literature review indicating that people allocate more study time to judged difficult than to judged-easy items--consistent with extant models of study-time allocation. However, typically, the materials were short, and participants had ample time for study. In contrast, in Experiment 1, when participants had insufficient time to study, they allocated more time to the judged-easy items than to the judged-difficult items, especially when expecting a test. In Experiment 2, when the materials were shorter, people allocated more study time to the judged-difficult materials. In Experiment 3, under high time pressure, people preferred studying judged-easy sonnets; under moderate time pressure, they showed no preference. These results provide new evidence against extant theories of study-time allocation. PMID- 10682299 TI - Functional characteristics of auditory temporal-spatial short-term memory: evidence from serial order errors. AB - The functional characteristics of auditory temporal-spatial short-term memory were explored in 8 experiments in which the to-be-remembered stimuli were sequences of bursts of white noise presented in spatial locations separated in azimuth. Primacy and recency effects were observed in all experiments. A 10-s delay impaired recall for primacy and middle list items but not recency. This effect was shown not to depend on the response modality or on the incidence of omissions or repetitions. Verbal and nonverbal secondary tasks did not affect memory for auditory spatial sounds. Temporal errors rather than spatial errors predominated, suggesting that participants were engaged in a process of maintaining order. This pattern of results may reflect characteristics that serial recall has in common with verbal and spatial recall, but some are unique to the representation of memory for temporal-spatial auditory events. PMID- 10682300 TI - Role of study strategy in recall of mixed lists of common and rare words. AB - Experiment 1 confirmed previous findings that common words are more recallable than are rare words when the 2 kinds of words are presented in separate lists but not when they are presented in the same list. Experiment 2 showed much the same pattern when an orienting task was performed during word presentation. In Experiment 3 common words were found to be more recallable than rare words even for mixed lists when no warning was given of the memory test, although the effect was less pronounced than for pure lists. In Experiment 4 stronger measures were taken to preclude anticipation of the memory test, and the effect of word commonness was found to be just as pronounced with mixed lists as it was with pure lists. It was suggested that lists are studied in a way believed to optimize recall and that mixed lists foster a strategy of favoring the rare words. PMID- 10682301 TI - Implicit learning differences: a question of developmental level? AB - A. S. Reber's (1992) proposition that the implicit learning system should demonstrate invariance of intellectual level (IQ) was examined by comparing 20 children with intellectual disability (mean mental age [MA] = approximately 5.8 years) with 20 intellectually gifted children (mean MA = approximately 12.4 years) of similar chronological age (CA; approximately 9.5 years). Implicit learning was assessed using a task involving covariation of 2 incidental cues. Explicit learning was assessed using a task of similar logical structure. Contrary to the IQ-invariance proposition, implicit learning as well as explicit learning varied with intellectual level. A secondary aim was to distinguish the contributions of CA, IQ, and MA to implicit learning. This was done by combining the samples of children in the present study with 2 samples of younger and older children of average ability from a study by M. Maybery, M. Taylor, and A. O'Brien Malone (1995). Analyses showed that MA is critical to implicit learning. PMID- 10682302 TI - Psychiatric symptoms of inherited metabolic disease. AB - Inborn errors of metabolism often present with a variety of psychiatric symptoms. With improved diagnosis and treatment options, many patients have increased lifespans; consequently, issues of long-term quality of life are coming to the forefront. Mental health concerns are among these issues. To demonstrate the connection between the course of metabolic disease and its psychiatric manifestations, four different inborn errors of metabolism are reviewed: phenylketonuria, Wilson disease, acute intermittent porphyria, and metachromatic leukodystrophy. PMID- 10682303 TI - Brain phenylalanine concentration in the management of adults with phenylketonuria. AB - Diagnosis by newborn screening and the implementation of a phenylalanine restricted diet have resulted in normal neurological development in approximately 10,000 persons with phenylketonuria (PKU) in the United States. While it is accepted that a phenylalanine-restricted diet is necessary in childhood, the recommended concentration of phenylalanine in the blood varies. Clinicians now must make recommendations for adults with PKU who probably tolerate higher levels of phenylalanine than children. This factor, quality of life issues, the expense of the diet, and varying genetic and socioeconomic backgrounds, make the choice of dietary recommendations difficult. Molecular analysis of the mutations in PKU has provided insight but has not resulted in clear recommendations for phenylalanine concentration in the blood. Magnetic resonance imaging has provided the recognition that white-matter changes are present in PKU. However, owing to poor correlation of white-matter changes with clinical factors, analysis of white matter changes has not proved useful. We hypothesize that measurement of brain phenylalanine directly will aid in clinical decision making. Twenty-one subjects with PKU had blood and brain phenylalanine measured simultaneously. Fifteen were randomly selected, 2 were examined for clinical reasons and 4 exceptional patients were chosen because they had maintained high IQs, despite having high historic blood concentrations and having been off the diet for at least 10 years. The correlation of blood and brain phenylalanine is in general poor. However, the four exceptional patients all had relatively low concentrations of phenylalanine in their brains compared to their blood. We suggest that their good clinical status, despite high historic blood levels, is due to their comparatively low brain levels of phenylalanine. We further suggest that measurement of brain phenylalanine concentration is useful in the management of PKU patients. PMID- 10682304 TI - Treatment of late-onset nonketotic hyperglycinaemia: effectiveness of imipramine and benzoate. AB - We report a patient with late-onset nonketotic hyperglycinaemia managed with a sequential approach to drug therapy in placebo-controlled therapeutic trials. Partial response to low-protein diet and sodium benzoate and dramatic response to imipramine are demonstrated, with parental scores on the Developmental Behavioural Checklist falling from the 86th centile before treatment to normal with combined benzoate and imipramine therapy. PMID- 10682305 TI - Effect of sodium benzoate in the treatment of atypical nonketotic hyperglycinaemia. AB - A 6-month-old girl presented with hypotonia and mild psychomotor retardation. Subsequently, an atypical manifestation of a nonketotic hyperglycinaemia was diagnosed, confirmed by significantly reduced activity of the glycine cleavage system in the liver tissue. After the patient developed hypsarrhythmia and had a single cerebral seizure, treatment with both sodium benzoate and dextromethorphan was started. During the following year, the girl was free of seizures with improvement of the EEG activity and showed retarded but continuously progressing psychomotor development. At the age of 20 months she began to walk freely but had generalized muscular hypotonia and moderate mental retardation. Discontinuation of dextromethorphan medication after one year of treatment did not change the clinical and electroencephalographic status. However, after cessation of sodium benzoate therapy, epileptic activity in the EEG and behavioural changes occurred. These changes disappeared promptly after sodium benzoate therapy was reinstituted. Thus, this case of mild atypical nonketotic hyperglycinaemia with only moderate psychomotor retardation and without epilepsy benefited from treatment with sodium benzoate in terms of electroencephalographic and behavioural changes. PMID- 10682306 TI - Acylcarnitines in fibroblasts of patients with long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency and other fatty acid oxidation disorders. AB - Mitochondrial fatty acid oxidation disorders cause hypoglycaemia, hepatic dysfunction, myopathy, cardiomyopathy and encephalopathy. Despite their recognition for more than 15 years, diagnosis and treatment remain difficult. To help design rational diagnostic and therapeutic strategies, we studied the pathophysiology of accumulating metabolites in a whole-cell system. Acylcarnitines were quantified in cells and media of cultured fibroblasts after incubation with L-carnitine and fatty acids. Following incubation with palmitate, long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD)-deficient fibroblasts compared with controls showed elevation of hydroxypalmitoyl- and palmitoyl-carnitine and reduction of C10- and shorter acylcarnitines, and following incubation with linoleate an increase in C14:2-, C18:2- and hydroxy-C18:2- acylcarnitines and reduction in C10:1-acylcarnitines. Hydroxyacylcarnitines remained more intracellular compared to corresponding saturated acylcarnitines. Incubation with decanoate and octanoate showed absence of hydroxylated acylcarnitines and correction of secondary metabolic disturbances, suggesting that optimal treatment should include medium-chain triglycerides of these chain lengths. Fibroblasts of patients with other fatty acid oxidation disorders showed distinct elevations of disease-specific acylcarnitines. This acylcarnitine analysis allows the diagnosis of LCHAD deficiency and its differentiation from other fatty acid oxidation disorders, which can pose difficulties in vivo. The strategy has allowed in-depth analysis with different substrates, providing suggestions for the rational design of treatment trials. PMID- 10682307 TI - Dietary therapy in two patients with a mild form of sulphite oxidase deficiency. Evidence for clinical and biological improvement. AB - We report an attempt at dietetic therapy in two unrelated patients with isolated sulphite oxidase deficiency, with a mild clinical course and late onset of symptoms. In case 1, disease started at 15 months with an acute crisis of agitation, unexplained crying and restlessness following otitis. Case 2 was diagnosed at 10 months when she presented with slight motor delay and dislocation of lenses. In both cases, sulphite oxidase activity measured in fibroblasts was undetectable. Therapy consisted of a diet low in protein from natural foods (daily methionine intake 130-150 mg) and a synthetic amino acid mixture (50 g per day) without cystine and methionine (Xmet, Cys Maxamaid, SHS International Ltd). A comparison of clinical and biochemical parameters was made between the period before treatment and after 2 years of treatment. Restriction in protein and sulphur amino acids brought about a dramatic decrease of urinary thiosulphate and S-sulphocysteine. It also brought about a generalized hypoaminoacidaemia with a low plasma methionine and cystine in both patients. Furthermore, both patients grew normally with no signs of neurological deterioration, and there was evidence of progress in psychomotor development. PMID- 10682308 TI - Pharmacological and genetic modifications of somatic cholesterol do not substantially alter the course of CNS disease in Niemann-Pick C mice. AB - Niemann-Pick type C (NPC) is a neurodegenerative disorder with somatically altered cholesterol metabolism. The NPC1 gene has recently been cloned and shown to have sequences shared with known sterol-sensing proteins. We have used a mouse model with a disrupted Npc1 gene to study two cholesterol-lowering drugs (nifedipine and probucol) and the effects of introducing a null mutation in the low-density lipoprotein receptor (LDLR). Although these treatments significantly ameliorated liver cholesterol storage, little effect on the onset of neurological symptoms was found. PMID- 10682310 TI - The natural course of Gaucher disease in The Netherlands: implications for monitoring of disease manifestations. AB - This retrospective study in 20 untreated type I Gaucher disease patients shows that in Dutch patients clinical manifestations of Gaucher disease type I are progressive in the majority of patients, children as well as adults. This is in contrast with studies among Jewish patients. Our results emphasize the need for a regular follow-up to enable timely initiation of enzyme therapy. PMID- 10682309 TI - A non-glycosylated and functionally deficient mutant (N215H) of the sphingolipid activator protein B (SAP-B) in a novel case of metachromatic leukodystrophy (MLD). AB - The lysosomal degradation of sphingolipids with short oligosaccharide chains depends on small glycosylated non-enzymatic sphingolipid activator proteins (SAPs, saposins). Four of the five known SAPs, SAP-A, -B, -C and -D, are derived by proteolytic processing from a common precursor protein (SAP-precursor) that is encoded by a gene on chromosome 10 consisting of 15 exons and 14 introns. SAP-B is a non-specific glycolipid binding protein that stimulates in vitro the hydrolysis of about 20 glycolipids by different enzymes. In vivo SAP-B stimulates in particular the degradation of sulphatides by arylsulphatase A. So far, four different point mutations have been identified on the SAP-B domain of the SAP precursor gene. The mutations result in a loss of mature SAP-B, causing the lysosomal accumulation of sulphatides and other sphingolipids, resulting in variant forms of metachromatic leukodystrophy (MLD). Here we report on a patient with SAP-B deficiency that is caused by a new homoallelic point mutation that has been identified by mRNA and DNA analysis. A 643A > C transversion results in the exchange of asparagine 215 to histidine and eliminates the single glycosylation site of SAP-B. Metabolic labelling experiments showed that the mutation had no effect on the intracellular transport of the encoded precursor to the acidic compartments and its maturation in the patient's cells. All four SAPs (SAP-A to SAP-D) were detectable by immunochemical methods. SAP-B in the patient's cells was found to be slightly less stable than the protein in normal cells and corresponded in size to the deglycosylated form of the wild-type SAP-B. Feeding studies with non-glycosylated SAP-precursor, generating non-glycosylated SAP-B, showed that the loss of the carbohydrate chain reduced the intracellular activity of the protein significantly. The additional structural change of the patient's SAP-B, caused by the change of amino acid 215 from asparagine to histidine, presumably resulted in an almost completely inactive protein. PMID- 10682311 TI - Neurological deterioration in adult phenylketonuria. PMID- 10682312 TI - The glutamine paradox in a neonate with propionic acidaemia and severe hyperammonaemia. PMID- 10682313 TI - Protein, glucose and energy metabolism in Gaucher disease type I. PMID- 10682314 TI - Increased urine heparan and chondroitin sulphate excretion in patients with osteopetrosis. PMID- 10682315 TI - Pyridoxine-responsive nephrocalcinosis and glycolic aciduria in two siblings without hyperoxaluria and with normal alanine: glyoxalate aminotransferase activity. PMID- 10682316 TI - The target hypothesis as a "quantum leap" to gene mutations. PMID- 10682317 TI - Fukuyama-type congenital muscular dystrophy: the first human disease to be caused by an ancient retrotransposal integration. AB - Fukuyama-type congenital muscular dystrophy (FCMD), one of the most common autosomal recessive disorders in the Japanese population, is characterized by congenital muscular dystrophy in combination with cortical dysgenesis (micropolygyria). Recently we identified on chromosome 9q31 the gene responsible for FCMD, which encodes a novel 461 amino acid protein that we have termed fukutin. Most FCMD-bearing chromosomes (87%) derive from a single ancestral founder, whose mutation consisted of a 3-kb retrotransposal insertion in the 3' noncoding region of the fukutin gene. Two independent point mutations causing premature termination confirmed that that this gene is responsible for FCMD. FCMD is the first human disease to be caused by an ancient retrotransposal integration. Fukutin contains an amino-terminal signal sequence, which together with results from transfection experiments suggests that it is an extracellular protein. Discovery of the FCMD gene represents an important step toward greater understanding of the pathogenesis of muscular dystrophies and also of normal brain development. PMID- 10682318 TI - Apoptosis or necrosis for tumor immunotherapy: what's in a name? AB - Here we discuss how the mechanisms by which tumor cells are killed in vivo by gene transfer affects their immunogenicity. Our own work has shown that necrotic cell death induces immunological activation signals which recruit, load, activate and mature appropriate subsets of antigen-presenting cells. In contrast, for apoptotic cell death to be immunogenic, signals additional to cell death alone must be provided within the milieu of the dying tumor. Our conclusion is that the immunogenicity of tumor killing is determined by a combination of factors, including the mechanism of killing, the levels of cell death, the local environment that exists within the dying tumor and, as a result, the nature of the immune/scavenger cells which are present at the time of antigen release. Knowledge of how these factors can influence the immune system and lead to the breaking of tolerance to tumor-associated antigens, can potentially be exploited in the design of effective immunotherapies for cancer using gene transfer. PMID- 10682319 TI - Analysis of the genetic diversity of Helicobacter pylori: the tale of two genomes. AB - Infection with Helicobacter pylori has been linked to numerous severe gastroduodenal diseases including peptic ulcer and gastric cancer. Several techniques have been used to measure the genetic heterogeneity of H. pylori at several different levels and to determine whether there is any correlation with severity of disease. The availability of two completed genome sequences from unrelated strains (J99 and 26,695) has allowed an analysis of the level of diversity from a large-scale yet detailed perspective. Although the two chromosomes are organized differently in a limited number of discrete regions, the genome size and gene order of these two "high-virulence" (cagA+ and vacA+) H. pylori isolates was found to be highly similar. The regions of organizational difference are associated with insertion sequences, DNA restriction/modification genes, repeat sequences, or a combination of the above. A significant level of variation at the nucleotide level is seen across the genome, providing an explanation for why the nucleotide-based typing techniques have such high discriminatory power among independent H. pylori isolates. This nucleotide variation together with the organizational rearrangements appears to have provided an over-estimation of the gene order diversity of H. pylori as assessed by pulse-field gel electrophoresis. Functional assignments are assigned to approximately only 60% of the gene products in each strain, with one-half of the remaining gene products of unknown function having homologues in other bacteria, while the remainder appear to be H. pylori-specific. Between 6% and 7% of the coding capacity of each strain are genes that are absent from the other strain, with almost one-half of these strain-specific genes located in a single hypervariable region called the plasticity zone. The majority of the strain specific genes in each strain are also H. pylori-specific, with no homologues being identified in the public databases. Significantly, over one-half of the functionally assigned strain-specific genes in both H. pylori J99 and 26695 encode DNA restriction/modification enzymes. Analysis of the level of conservation between orthologues from the two strains indicates that the H. pylori specific genes have a lower level of conservation than those orthologues to which a putative function can be assigned. The plasticity zone represents one of several regions across each genome that is comprised of lower (G+C)% content DNA, some of which has been detected in self-replicating plasmids, suggesting that both horizontal transfer from other species and plasmid integration are responsible for the strain-specific diversity at this locus. These analyses have yielded results with important implications for understanding the genetic diversity of H. pylori and its associated diseases, and imply a need to reassess the respective roles of bacterial and host factors in H. pylori associated diseases. PMID- 10682320 TI - Analysis of differentially regulated mRNAs in monocytic cells induced by in vitro stimulation. AB - Macrophages are known to be effector cells in several granulomatous disorders. However, little is known about granuloma-associated up- or downregulation of genes in these cells. Differential display reverse transcription polymerase chain reaction (DDRT-PCR) is an attractive method for the detection of differentially expressed genes. Although this method entails a number of drawbacks, its application to rare and limited amounts of clinical samples is still convenient. In this study, we introduce a screening procedure for detecting differentially regulated sequence tags in samples of patients suffering from granulomatous diseases. We applied DDRT-PCR in a multiple and complex comparison of expressed sequence tags in response to various granuloma-associated stimuli. The histiocytic cell line U937 was used as a model. The cells had been stimulated with granuloma-associated agents such as Mycobacterium tuberculosis, BeSO4, lipopolysaccharide, or HgS and unspecific stimuli such as phorbol myristate acetate, phytohemagglutinin, Zymosan, and Latex. Comparative analysis of 2237 sequence tags obtained from 55 primer combinations revealed 22.4% differentially amplified PCR products. Notably, only 8.0% of the differentially expressed sequence tags showed an association restricted to in vitro cultivation in the presence of M. tuberculosis, lipopolysaccharide, BeSO4, and/or HgS, while 1.0 1.9% of the tags were altered exclusively as a consequence of stimulation with one of the granuloma-associated agents. Our data provide evidence that this strategy may function as a preselection for appropriate primer combinations to discover sequence tags which could be specifically associated with granulomatous disorders. This approach could shorten laborious screening, save consumption of valuable and rare samples, and could reduce the number of false-positive results. PMID- 10682321 TI - Relationship of angiotensin-converting enzyme gene polymorphism to carotid wall thickness in middle-aged men. AB - The insertion/deletion (I/D) polymorphism of the human angiotensin-converting enzyme (ACE) gene is a major determinant of circulating ACE levels. The D allele has been suggested to be a potent risk factor for coronary artery disease; however, the effect of the ACE gene on carotid atherosclerosis remains controversial. We therefore studied the relationship between the ACE gene I/D polymorphism and carotid artery intima-media thickness (IMT). A random sample of 300 men aged 50-59 years living in southern Finland were selected, and 233 agreed to participate (74%). Data were collected in 219 subjects. Quantitative B-mode ultrasonography was used to measure the maximum near and far wall IMT of right and left common, bifurcation, and internal carotid artery. The mean maximum IMT (overall mean) was calculated as the mean of 12 maximum IMTs at 12 standard sites. Patients with an IMT higher than 1.7 mm in at least one of 12 standard sites were assumed to have carotid atherosclerosis. The I/D polymorphism was determined by polymerase chain reaction. Overestimation of the frequency of the DD genotype was eliminated by insertion-specific primer and the inclusion of 5% dimethylsulfoxide. No significant differences were found in carotid wall thickness between the three genotypes; the overall mean IMT were 1.18 +/- 0.30, 1.22 +/- 0.24, and 1.08 +/- 0.40 mm in genotypes of II, ID, and DD, respectively. Similarly, the ACE genotypes and allele frequencies did not differ significantly between the subjects with and those without carotid atherosclerosis. There was no association in the subgroups among only nonsmoking subjects or subjects without chronic medication. The present data indicate that the I/D polymorphism of the ACE gene is not related to carotid IMT and is unlikely to play a major role in carotid atherosclerosis. PMID- 10682322 TI - Efficient gene delivery into adult cardiomyocytes by recombinant Sindbis virus. AB - Somatic gene therapy as a potential strategy for the treatment of myocardial diseases relies on an efficient gene transfer into cardiac muscle cells. The difficulty of delivering genes into adult cardiomyocytes exists not only in vivo but also in primary culture systems. Therefore, possibilities for ex vivo gene transfer and the in vitro study of physiological processes by reverse genetics are limited. We investigated the potential of an alphavirus-based vector system to transduce adult rat cardiomyocytes (ARC) in culture using a replication deficient Sindbis virus (SIN) encoding beta-galactosidase (SIN-LacZ). Transduction efficiency depended on the virus concentration used, with expression of the reporter gene being detectable in up to 80% of cultured ARC as early as 24 h after infection. We observed a remarkably lower cytotoxicity of this viral vector in ARC than in other cells such as fibroblasts and neonatal cardiomyocytes. Additionally, no perceptible changes in the morphology of the nuclei or cytoskeleton were found in ARC 48 h after infection with SIN-LacZ. We conclude that SIN vectors are useful for gene delivery into adult cardiomyocytes and believe that improved versions of this viral system may be useful for cardiovascular gene therapy in the future. PMID- 10682324 TI - Dosage schedule for dietary fluoride supplements. Proceedings of a workshop. Chicago, Illinois, USA. January 31-February 1, 1994. PMID- 10682323 TI - The expression and role of insulin-like growth factor II in malignant hemangiopericytomas. AB - Hemangiopericytoma is a rare soft tissue tumor originating from contractile pericapillary pericytes. To address the issue of molecular genetic events that participate in genesis and progression of hemangiopericytoma we analyzed insulin like growth factor (IGF) II and IGF I receptor in 29 tumors collected from a human tumor bank network. Seven of these tumors were associated with severe hypoglycemia; six were retroperitoneal and one was located in the leg. Of 22 tumors tested 12 (54.5%) exhibited IGF II mRNA, while almost 90% (17 of 19) of hemangiopericytomas exhibited IGF I receptor mRNA. Sera from some patients whose tumors expressed IGF II mRNA contained elevated levels of IGF II. Removal of the tumor eliminated most of the IGF II immunoreactivity from the sera. The potential role of IGF II as a growth-promoting factor was examined on three malignant primary hemangiopericytoma cell cultures. Extracellular addition of IGF II significantly enhanced cell proliferation in a dose-dependent manner. Antisense oligodeoxynucleotides that specifically inhibit IGF II mRNA, at a concentration of 40 or 80 micrograms/ml, inhibited the growth of hemangiopericytoma cells significantly, by 40%. Simultaneous administration of antisense deoxyoligonucleotides to both IGF II and IGF I receptor inhibited tumor cell proliferation by even 80%. Our data suggest that tumor cells produce IGF II, and that this in turn stimulates their proliferation by autocrine mechanisms. PMID- 10682325 TI - The role of dietary fluoride supplements in caries prevention. AB - Nearly all dental researchers and public health authorities agree that fluoride supplements are highly effective in reducing dental caries in primary and permanent teeth, that benefits to all teeth are greater when administration begins at 2 years of age or younger, that both preeruptive and posteruptive exposure is important in imparting cariostatic benefits, that effectiveness is neither enhanced nor reduced by their being combined with vitamins, and that benefits to the offspring of pregnant women who take supplements are uncertain. Several studies show that fluoride supplements delivered in school-based programs effectively reduce dental caries, and benefits are greater to teeth that receive preeruptive exposure in addition to posteruptive exposure. Many parents who, for a variety of reasons, did not administer fluoride supplements at home will enroll their children in school-based fluoride tablet programs. Effectiveness of fluoride supplements today is undoubtedly smaller than observed previously because of dilution and diffusion effects from other fluoride sources; nevertheless, they still have the same potential efficacy. It is apparent that the current ADA dosage schedule is too high and requires modification; however, the availability of this known-to-be-effective regimen should not be eliminated or restricted. PMID- 10682326 TI - Total fluoride intake and implications for dietary fluoride supplementation. AB - This paper reviews the history and validity of recommended "optimal" levels of systemic fluoride intake and the available information on levels of fluoride intake in young children from foods and beverages (including water), dentifrices, dietary fluoride supplements, mouthrinses, and gels. Most of the studies emphasize the substantial variation in ingestion among individuals. Often, a substantial proportion of individuals received fluoride well beyond the mean exposure reported in the study. Limitations in the existing data make it difficult to determine the total distribution of fluoride intake from all sources. Therefore, hypothetical combinations of possible daily fluoride intake from the three main sources (diet, dentifrices, and supplements) are presented for those aged 6, 12, 24, and 36 months, with associated mean intake per kg body weight. Findings suggest that some children exceed the "optimal" level of fluoride intake from single sources alone, while others can from a combination of sources. Moreover, if current recommended "optimal" levels, which have been derived on an empirical basis, are actually lower than what has been quoted in the literature, then more children could be ingesting excessive amounts of fluoride, which could increase their risk of developing objectionable dental fluorosis. The variation and complexity of fluoride ingestion from all sources should be considered in the evaluation of recommendations for use of dietary fluoride supplements. PMID- 10682327 TI - Fluoride metabolism and excretion in children. AB - This paper compares fluoride pharmacokinetics (plasma, renal, and extrarenal clearances) and metabolic balances in healthy infants or children with those in young or middle-aged adults. Regardless of age, the removal of fluoride from the intra- and extracellular body fluids occurs almost exclusively by uptake in calcified tissues and excretion in the urine. While there can be considerable differences among individuals, the rates at which fluoride is cleared from plasma by calcified tissues and the kidneys in adults are approximately equal. The calcified tissue clearance of fluoride from plasma in children is substantially higher than that by the kidneys. This is due to the greater surface area of the loosely organized crystallites in the developing calcified tissues during growth. Thus, the balance of fluoride (total intake minus total excretion) is typically higher in children than in adults, but it can be positive or negative at any age. Positive balance occurs when chronic fluoride intake is sufficient to prevent plasma concentrations from declining. When positive, the fluoride content of the calcified tissues, which contain more than 99 percent of the body's fluoride, tends to gradually increase. Negative balance, which indicates net mobilization of fluoride from calcified tissues, can occur when plasma concentrations decline due to a reduction in the level of fluoride intake. PMID- 10682328 TI - Fluoride intake by infants. AB - Many infants are fully or partially breast fed during the early months of life; however, the percentage of such infants decreases to about 30 percent by 4 months of age. The majority of US infants are fed formulas for most of the first 10 months of life. Although fluoride (F) intakes by fully breast-fed infants are low, F intakes by partially breast-fed infants and by formula-fed infants are highly variable, depending primarily on the F content of the water used to dilute concentrated liquid or powdered infant formula products. In communities with F content of the drinking water less than 0.3 ppm, F consumption by many infants will be 30 to 40 micrograms.kg-1.d-1. The addition of a F supplement of 0.25 mg/d for a 4 kg infant would increase the F intake by 63 micrograms.kg-1.d-1, resulting in a total intake of about 100 micrograms.kg-1.d-1, an intake in the range believed to be associated with development of fluorosis of the permanent teeth. However, for the US infant population generally, many fewer infants are exposed to high F intakes from formula plus a supplement (recommended only for communities with water providing less than 0.3 ppm F) than from formula alone in communities with F content of 1 ppm in the drinking water. In assessing the possible effects of F intake during infancy on development of fluorosis, it is important to recognize that infant feeding practices have changed greatly during the past 30 years. In the 1960s, most infants over 4 months of age were fed fresh cow's milk and intakes of F were therefore low. By the mid 1970s a trend toward more extended feeding of formula was evident and this trend has continued into the 1990s. Prolonged exposure to high intakes of fluoride during infancy is much more common now than in the past. PMID- 10682329 TI - The differential diagnosis of fluorosis. AB - Following the introduction of the first fluorosis index by Dean, a series of fluorosis indexes were introduced. While they may differ in the specific way fluorosis is categorized, they all nevertheless use the same underlying diagnostic signs--originally described by Dean, Black, and McKay--that were causally linked to the development of enamel in areas with above-optimum fluoride in the drinking water. Underlying the various fluorosis indexes is the belief that specific clinical diagnostic criteria, based upon established clinical signs, can be utilized to differentiate fluorotic from nonfluorotic enamel opacities. These criteria repeatedly have been substantiated in studies in which the presence of enamel fluorosis, identified by clinical differential diagnosis, has been associated with fluoride exposure history. Further, to whatever extent nonfluorotic opacities have been misdiagnosed as fluorosis, observed estimates of association derived from analytical studies will have been underestimated. PMID- 10682330 TI - The prevalence and severity of enamel fluorosis in North American children. AB - The question considered in this review is the extent to which changes in the prevalence or severity of enamel fluorosis have occurred over the last half century. Emphasis is given to a review of those studies in which subjects are drinking water that is fluoride deficient and those in which subjects are drinking optimally fluoridated water, either adjusted or natural. Trends in fluorosis were examined using two definitions of fluorosis (definite and any signs) and three types of comparisons--comparisons of pooled estimates from all available studies that include data from different communities and time periods, comparisons of estimates from the same communities at different times, and comparisons of estimates from selected studies in the early years of fluorosis research with results of the US National Fluorosis Survey done by the National Institute of Dental Research. A clear increase in fluorosis among populations drinking community water that contains less than 0.3 ppm fluoride was found. Results of the comparisons using studies with Dean's Index pooled at different time points, comparisons in the same communities over time, and comparisons of prevalence found in selected communities before fluoride was widely available with the National Fluorosis Survey all support this conclusion. An increase in the prevalence of fluorosis in those drinking optimally fluoridated water likely has occurred as well; however, evidence for such a trend is not as clear as for fluoride deficient communities because of mixed results depending on the type of comparison. The majority of fluorosis cases continue to be mild and seem of little esthetic consequence for most of the public or dental profession. But a few cases of more severe fluorosis can be found now in some communities. Because the prevalence of fluorosis is now higher than 50 years ago, we can conclude that fluoride availability to the developing enamel during critical periods when enamel is at risk of fluorosis has increased in North American children. PMID- 10682331 TI - Mechanism and timing of fluoride effects on developing enamel. AB - Fluoride appears to specifically interact with mineralizing tissues, causing an alteration of the mineralization process. In enamel, fluorosis results in a subsurface hypomineralization. This hypomineralized enamel appears to be directly related to a delay in the removal of amelogenins at the early-maturation stage of enamel formation. The specific cause for this delay is not known, although existing evidence points to reduced proteolytic activity of proteinases that hydrolyze amelogenin. This delay in hydrolysis of amelogenins could be due to a direct effect of fluoride on proteinase secretion or proteolytic activity, or to a reduced effectiveness of the proteinase due to other changes in the protein or mineral of the fluorosed enamel matrix. The formation of dental fluorosis is highly dependent on the dose, duration, and timing of fluoride exposure. The early-maturation stage of enamel formation appears to be particularly sensitive to the effects of fluoride on enamel formation. Although the risk of enamel fluorosis is minimal with exposure only during the secretory stage, this risk is greatest when exposure occurs in both secretory and maturation stages of enamel formation. The risk of fluorosis appears to be best related to the total cumulative fluoride exposure to the developing dentition. PMID- 10682332 TI - Overview of the history and current status of fluoride supplementation schedules. AB - Clinical trials of dietary fluoride supplements began in the 1940s in an effort to bring the benefits of fluoride to those who did not receive it through their drinking water. Following the early success of these trials, the Council on Dental Therapeutics of the American Dental Association (ADA) published its first recommendations for fluoride supplementation in 1958. The American Academy of Pediatrics (AAP) followed with its own recommendations in 1972. During the 1970s a variety of alternative schedules appeared in the literature, most in reaction to the findings of unexpectedly high levels of enamel fluorosis in children being supplemented with the AAP schedule. In 1979 the ADA and AAP agreed on essentially identical schedules. During the 1980s, however, the prevalence of enamel fluorosis continued to increase, and fluoride supplements were found in some studies to be a risk factor for fluorosis. This finding prompted another round of dosage schedule recommendations in the early 1990s. This paper presents a history of fluoride dosage recommendations and reviews the recent proposals for reducing supplement dosage. PMID- 10682333 TI - The case for retaining the current supplementation schedule. AB - Following ingestion of dietary fluoride, microquantities of fluoride return to the mouth in saliva, but in quantities large enough to facilitate the maintenance and reparative functions of enamel. Dietary fluoride supplements alone are unlikely to be the cause of the reported increase in fluorosis. Compliance continues to be extremely poor and few children use supplements for more than a year and a half. The amount of background fluoride resulting from dietary fluoride supplements appears to be very small. Considering the almost ubiquitous presence of fluoride dentifrice and the strong possibility of additional unintentional fluoride ingestion from many sources, the present fluorosis data is too amorphous to use as a basis for making reasonable risk/benefit evaluations. Very mild and mild fluorosis is not a serious problem for either the clinician or the patient. By altering the present recommended dosage we may deprive children from receiving a proven effective dose. One cannot make a risk/benefit decision concerning an esthetic problem without involving the patient's perception as well as the caries score. The apparent severity of the milder forms of fluorosis lessens with age and a community fluorosis index should be used only on populations who are older than 15 years. PMID- 10682334 TI - The case for reducing the current Council on Dental Therapeutics fluoride supplementation schedule. AB - The milder forms of dental fluorosis have increased in prevalence since the original epidemiologic surveys of the 1930s. Most studies of fluorosis have identified the use of supplements as a major risk factor. Fluorosis could be prevented, in part, by stopping the improper prescription of fluoride supplements in optimally fluoridated areas and by lowering the dosage currently recommended by the Council on Dental Therapeutics supplemental fluoride schedule. At a 1991 workshop at the University of North Carolina, five alternatives to the present ADA Council on Dental Therapeutics schedule were suggested; however, no consensus on dosage was reached. Recently, the Federation Dentaire International adopted a dosage schedule of 0.25 mg F from birth to 3 years of age, 0.5 mg F from 3 to 5 years, and 1 mg F thereafter. At a 1992 Canadian workshop it was proposed that supplements should not be started until age 3, should be given only to those "at high risk" of caries, and only 0.25 mg F should be prescribed from 3 to 5 years of age. Similarly, in some European countries supplements are not recommended until 3 years, at which time 0.5 mg F is prescribed, but only "for children at risk." Australia is considering a dosage schedule starting with 0.25 mg F at 6 months, again only for those "particularly at risk of caries." Serious problems exist in limiting fluoride supplementation only to high-caries-risk children because they are not easily identifiable at a young age. Ideally, a dosage schedule should be based on body surface area or weight rather than simply age, and supplements should be in the form of lozenges for children over 2 years of age. A reduced fluoride supplement dosage schedule is proposed. PMID- 10682335 TI - The case for eliminating the use of dietary fluoride supplements for young children. AB - Fluoride supplements have been used for years to prevent dental caries; nevertheless, there are three reasons why their use is inappropriate today among infants and young children in the United States. Evidence for the efficacy of fluoride supplements when used from birth or soon after is weak, supplements are a risk factor for dental fluorosis, and fluoride has little preeruptive effect in caries prevention. While there are many reports on the caries-preventive efficacy of supplements, few meet standards for acceptability as clinical trials, and those that do have tested chewable tablets or lozenges under supervision in school-aged children. North American children today are exposed to fluoride from many sources--drinking water, toothpaste, gels, rinses, and in processed foods and beverages. The additional cariostatic benefits that accrue from using supplements are marginal at best, while there is strong risk of fluorosis when young children use supplements. Available evidence suggests that the public is more aware of the milder forms of fluorosis than was previously thought; thus, it is prudent for caries-preventive policies to aim to maximizing caries reductions while minimizing the risk of fluorosis. It is therefore concluded that the risks of using supplements in infants and young children outweigh the benefits. Because alternative forms of fluoride for high-risk individuals exist, fluoride supplements should no longer be used for young children in North America. PMID- 10682336 TI - Challenges of and strategies for changing prescribing practices of health care providers. AB - Problems related to inappropriate prescribing practices of physicians in general are well recognized. Dietary fluoride supplements have been implicated as one of the contributing factors in an increase in dental fluorosis. Inappropriate prescribing practices of providers have been cited as a major factor in this implication. Numerous studies of physicians and dentists have documented a lack of knowledge and inappropriate prescribing practices regarding fluoride supplements. The purpose of this paper is to identify barriers to changing fluoride-prescribing practices of health care providers and to suggest strategies for implementing change. To increase optimal and appropriate use of fluoride supplements, educational interventions are necessary for all user groups--detail men and women, physicians, dentists, pharmacists, nurse practitioners, dental hygienists, and the public. In addition, environmental supports for the educational activities in the form of policy, regulation, standards of care, and guidelines are recommended for consideration. PMID- 10682337 TI - Testing for symmetry in the conditional discriminations of language-trained chimpanzees. AB - If subjects are taught to match Stimulus A to B and then, without further training, match B to A, they have passed a test of symmetry. It has been suggested that non-humans' lack of success on symmetry tests might be overcome by giving them a history of symmetry exemplar training, that is, by directly teaching a large number of conditional relations (e.g., AB, CD, EF,...) and also directly training the "reverse" of these relations (e.g., BA, DC, FE,...). The chimpanzee subjects of the present study, Sherman, Austin, and Lana, had already received extensive symmetry exemplar training as a result of attempts to teach a selection-based language system of lexigrams. The present study systematically subjected 2 of these chimps (Sherman and Lana), for the first time, to standard symmetry tests in controlled conditions. Both chimps failed the tests, even when their correct responses on test trials were reinforced. The findings do not support the exemplar training hypothesis, and cast doubt upon whether the chimps can pass tests of stimulus equivalence. PMID- 10682338 TI - Falsification of matching theory's account of single-alternative responding: Herrnstein's k varies with sucrose concentration. AB - Eight rats pressed levers for varying concentrations of sucrose in water under eight variable-interval schedules that specified a wide range of reinforcement rate. Herrnstein's (1970) hyperbolic equation described the relation between reinforcement and responding well. Although the y asymptote, k, of the hyperbola appeared roughly constant over conditions that approximated conditions used by Heyman and Monaghan (1994), k varied when lower concentration solutions were included. Advances in matching theory that reflect asymmetries between response alternatives and insensitive responding were incorporated into Herrnstein's equation. After fitting the modified equation to the data, Herrnstein's k also increased. The results suggest that variation in k can be detected under a sufficiently wide range of reinforcer magnitudes, and they also suggest that matching theory's account of response strength is false. The results support qualitative predictions made by linear system theory. PMID- 10682339 TI - Three predictions of the economic concept of unit price in a choice context. AB - Economic theory makes three predictions about consumption and response output in a choice situation: (a) When plotted on logarithmic coordinates, total consumption (i.e., summed across concurrent sources of reinforcement) should be a positively decelerating function, and total response output should be a bitonic function of unit price increases; (b) total consumption and response output should be determined by the value of the unit price ratio, independent of its cost and benefit components; and (c) when a reinforcer is available at the same unit price across all sources of reinforcement, consumption should be equal between these sources. These predictions were assessed in human cigarette smokers who earned cigarette puffs in a two-choice situation at a range of unit prices. In some sessions, smokers chose between different amounts of puffs, both available at identical unit prices. Individual subjects' data supported the first two predictions but failed to support the third. Instead, at low unit prices, the relatively larger reinforcer (and larger response requirement) was preferred, whereas at high unit prices, the smaller reinforcer (and smaller response requirement) was preferred. An expansion of unit price is proposed in which handling costs and the discounted value of reinforcers available according to ratio schedules are incorporated. PMID- 10682340 TI - Travel time and concurrent-schedule choice: retrospective versus prospective control. AB - Six pigeons were trained on concurrent variable-interval schedules in which two different travel times between alternatives, 4.5 and 0.5 s, were randomly arranged. In Part 1, the next travel time was signaled while the subjects were responding on each alternative. Generalized matching analyses of performance in the presence of the two travel-time signals showed significantly higher response and time sensitivity when the longer travel time was signaled compared to when the shorter time was signaled. When the data were analyzed as a function of the previous travel time, there were no differences in sensitivity. Dwell times on the alternatives were consistently longer in the presence of the stimulus that signaled the longer travel time than they were in the presence of the stimulus that signaled the shorter travel time. These results are in accord with a recent quantitative account of the effects of travel time. In Part 2, no signals indicating the next travel time were given. When these data were analyzed as a function of the previous travel time, time-allocation sensitivity after the 4.5-s travel time was significantly greater than that after the 0.5-s travel time, but no such difference was found for response allocation. Dwell times were also longer when the previous travel time had been longer. PMID- 10682341 TI - Choice between constant and variable alternatives by rats: effects of different reinforcer amounts and energy budgets. AB - Two experiments, using rats as subjects, investigated the effect of different reinforcer amounts and energy budgets on choice between constant and variable alternatives under a closed economy. Rats were housed in the chamber and were exposed to a modified concurrent-chains schedule in which the choice phase was separated from a rest phase during which the rats could engage in other activities. In the choice phase, a single variable-interval schedule arranged entry into one of two equal terminal links (fixed-interval schedules). The constant terminal link ended with the delivery of a fixed number of food pellets (two or three, depending on the condition), whereas the variable terminal link ended with a variable number of food pellets (means of two or three, depending on the condition). Energy budget was defined as positive when body weights were over 90% of free-feeding weights, and as negative when they were under 80% of free feeding weights. The different body weights were produced by varying the duration of the equal terminal-link schedules within daily 3-hr sessions. In Experiment 1, rats chose between a constant and a variable three pellets under both energy budgets. Rats preferred the constant three pellets more under the positive energy budget, whereas they were indifferent under the negative energy budget. In Experiment 2, rats chose between a constant three pellets and a variable two pellets, and chose between a constant two pellets and a variable three pellets under both energy budgets. The rats strongly preferred the constant three pellets over the variable two pellets under both energy budgets. In contrast, rats preferred the variable three pellets over the constant two pellets only under the negative energy budget, whereas they were indifferent under the positive energy budget. These results indicate that rats choices are sensitive to the difference in reinforcer amounts and to the energy budgets defined by the level of body weight. The present results are consistent with those obtained with small granivorous birds as well as with the predictions of a recent risk-sensitive foraging theory. PMID- 10682342 TI - Pigeons' choices between fixed-ratio and linear or geometric escalating schedules. AB - Four related procedures provided a basis for comparing the linear-optimality principle with a principle based on the sums of reciprocals of distances to reinforcement, and to explore the generality of the sums-of-reciprocals principle as a description of choice patterns in situations of diminishing returns. The procedures all arranged choices between fixed-ratio schedules and progressive ratio schedules, which escalated with each consecutive choice. In contrast to previous work that involved constant ratio increments, two sets of procedures in this study involved relatively small increments that are similar to the early values when a progressive schedule is increasing proportionally. The remaining two sets of procedures examined progressive schedules with proportional increments. In addition, the initial value of the progressive alternative was manipulated to determine its effects on patterns of choice with both linear and proportional types of escalation. With the exception of one phase, regardless of the initial/reset value and the patterns of escalation, patterns of choice with pigeons were well characterized by the sums-of-reciprocals principle. This supports previous research with pigeons using fixed-increment progressive schedules, as well as situations in which the progressive schedule increased by constant proportions instead of by constant increments. The findings are attributed to the feature of this averaging technique whereby it differentially values reinforcers based on their relative proximity to a particular choice point. PMID- 10682343 TI - Drug discrimination in rats under concurrent variable-interval variable-interval schedules. AB - Eight rats were trained to discriminate pentobarbital from saline under a concurrent variable-interval (VI) VI schedule, on which responses on the pentobarbital-biased lever after pentobarbital were reinforced under VI 20 s and responses on the saline-biased lever were reinforced under VI 80 s. After saline, the reinforcement contingencies programmed on the two levers were reversed. The rats made 62.3% of their responses on the pentobarbital-biased lever after pentobarbital and 72.2% on the saline-biased lever after saline, both of which are lower than predicted by the matching law. When the schedule was changed to concurrent VI 50 s VI 50 s for test sessions with saline and the training dose of pentobarbital, responding on the pentobarbital-biased lever after the training dose of pentobarbital and on the saline-biased lever after saline became nearly equal, even during the first 2 min of the session, suggesting that the presence or absence of the training drug was exerting minimal control over responding and making the determination of dose-effect relations of drugs difficult to interpret. When the pentobarbital dose-response curve was determined under the concurrent VI 50-s VI 50-s schedule, responding was fairly evenly distributed on both levers for most rats. Therefore, 6 additional rats were trained to respond under a concurrent VI 60-s VI 240-s schedule. Under this schedule, the rats made 62.6% of their responses on the pentobarbital-biased lever after pentobarbital and 73.5% of their responses on the saline-biased lever after saline, which also is lower than the percentages predicted by perfect matching. When the schedule was changed to a concurrent VI 150-s VI 150-s schedule for 5-min test sessions with additional drugs, the presence or absence of pentobarbital continued to control responding in most rats, and it was possible to generate graded dose response curves for pentobarbital and other drugs using the data from these 5-min sessions. The dose-response curves generated under these conditions were similar to the dose-response curves generated using other reinforcement schedules and other species. PMID- 10682344 TI - Highly purified thermo-stable oxygen-evolving photosystem II core complex from the thermophilic cyanobacterium Synechococcus elongatus having His-tagged CP43. AB - The carboxyl terminus of the CP43 subunit of photosystem II (PSII) in the thermophilic cyanobacterium, Synechococcus elongatus, was genetically tagged with six consecutive histidine residues to create a metal binding site on the PSII supramolecular complex. The histidine-tagging enabled rapid isolation of an intact cyanobacterial PSII core complex from dodecyl maltoside-solubilized thylakoids by a simple one-step Ni(2+)-affinity column chromatography. The isolated core complex was in a dimeric form with a molecular mass of about 580 kDa, consisting of five major intrinsic membrane proteins (CP47, CP43, D1, D2 and cytochrome b-559), three extrinsic proteins (33 kDa, 12 kDa, and cytochrome c 550), and a few low molecular mass membrane proteins, and evolved oxygen at a rate as high as 3,400 mumol (mg Chl)-1 h-1 at 45 degrees C with ferricyanide as an electron acceptor. The core complex emitted thermoluminescence B2-, B1- and Q bands arising from S2QB-, S3QB- and S2QA- charge recombinations at respective emission temperatures of 45, 38 and 20 degrees C, all of which were higher by about 15 degrees C as compared with those in mesophilic spinach BBY membranes. These results indicated that the isolated core complex well retained the intact properties of thermoluminescence of thermophilic cyanobacterial cells, the deeper stabilization of PSII charge pairs. The isolated complex was extremely stable in terms of both protein composition and function, exhibiting no release of extrinsic proteins, no proteolytic degradation in any of its subunits, accompanied by only a slight (less than 10%) loss in oxygen evolution, after dark incubation at 20 degrees C for 8 d. These properties of the thermophilic PSII core complex are highly useful for various types of studies on PSII. PMID- 10682345 TI - Molecular cloning of a defense-response-related cytochrome P450 gene from tobacco. AB - Plant defenses against pathogen attack involve a series of inducible responses that contribute to resistance. Tobacco leaves injected with HWC (hyphal wall components prepared from Phytophthora infestans) elicitor showed typical defense responses, including the induction of localized necrosis and the accumulation of pathogenesis-related proteins. In order to elucidate the molecular mechanisms by which plant defense systems are activated, we screened tobacco plants for genes differentially expressed in response to HWC. We performed differential screening by RT-PCR with random primers and obtained PCR products specific to HWC-treated leaf RNA. Northern hybridization using the PCR products as probes confirmed that one transcript was actually induced by HWC treatment. As the deduced amino acid sequence of this clone showed the highest degree of similarity to elicitor induced soybean cytochrome P450 CYP82A4, it was designated CYP82E1. The expression of CYP82E1 was strongly induced in tobacco by the soybean pathogen Pseudomonas syringae pv. glycinea (nonpathogenic on tobacco), but it was activated only slightly and in a delayed fashion by the tobacco pathogen P. syringae pv. tabaci (pathogenic on tobacco), implying that the product of CYP82E1 may be involved in disease resistance in tobacco. PMID- 10682346 TI - Localization of the Ca(2+)-binding protein, Bra r 1, in anthers and pollen tubes. AB - Calcium plays an essential role during pollen development and pollen tube growth, and several Ca(2+)-binding proteins are expressed in anthers. We have previously reported that Brassica pollen allergens encoded by Bra r 1 and Bra r 2 show sequence similarities to Ca(2+)-binding proteins [Toriyama et al. (1995) Plant Mol. Biol. 29: 1157]. Herein, we report that both genes are expressed in the diploid tapetum and haploid microspores, as detected by in situ RNA hybridization. Immunoblot analysis revealed that Bra r 1 and Bra r 2 were accumulated in anthers during pollen development. When pollen grains were suspended in an aqueous solution, both proteins were mainly detected in the pollen extracellular fraction, indicating that Bra r 1 and Bra r 2 are released from the pollen upon hydration. Localization of Bra r 1 was further investigated in sections of anthers and pollen tubes. Bra r 1 was detected in the tapetum, microspores and pollen grains. In longitudinal sections of cross-pollinated pistils. Bra r 1 was detected throughout pollen tubes elongating in transmitting tissue. These findings suggest that Bra r 1 may be involved in pollen-pistil interaction and pollen tube growth. PMID- 10682347 TI - Developmental transitions and dynamics of the cortical ER of Arabidopsis cells seen with green fluorescent protein. AB - Arabidopsis thaliana plants were stably transformed with DNA encoding green fluorescent protein and with sequences ensuring retention in the endoplasmic reticulum (ER). Confocal laser scanning microscopy shows fluorescent ER in many cells of seedlings so allowing developmental changes to be documented. The arrangement of the cortical ER changes as cells mature in the hypocotyl and root epidermis. In the root, cells that have completed expansion have reticulate cortical ER resembling the ER described in many previous studies. Expanding cells, however, show extensive perforated sheets of cortical ER which transform quite abruptly into a loose reticulum at the basipetal end of the elongation zone. The reticulum compacts in trichoblasts beginning at sites where root hairs are about to emerge. The compacted form is maintained throughout the hair until growth ceases and the open reticulate form returns. All forms of cortical ER are dynamic and we use a color overlay method to distinguish stable and moving structures in a single composite image. Reticulate ER continuously rearranges its polygonal layout and perforations move and change their shape in the ER sheets of younger cells. ER deeper in the cell (i.e. not close to the plasma membrane) moves more actively so that almost no tubules remain stable even over short periods of less than one minute. The function of the perforated sheets of cortical ER present in growing cells is unknown. PMID- 10682348 TI - Identification and expression of cotton (Gossypium hirsutum L.) plastidial carbonic anhydrase. AB - Four carbonic anhydrase (CA) cDNA clones were isolated from a 48 h dark-grown cotton (Gossypium hirsutum L.) seedling cDNA library. Nucleotide sequence analysis revealed two different CA isoforms designated GhCA1 and GhCA2. The encoded polypeptides possess N-terminal serine/threonine-rich regions indicative of plastid transit peptides, and approximately 80% sequence identity to other plant plastidial beta-CAs. The GhCA1 cDNA encodes a nearly complete preprotein of 323 amino acids with a molecular mass of 34.9 kDa and a predicted mature protein of 224 amino acids with a molecular mass of 24.3 kDa. Eleven nucleotide differences within ORFs of GhCA1 and GhCA2 result in 5 conservative amino acid substitutions. The 3' GhCA2 untranslated region contains five additional substitutions and one single nucleotide addition. GhCA1 clones, nearly full length or with 70% of the transit peptide deleted, were expressed as LacZ alpha fusion proteins in E. coli. Lysates of these strains contained 9-fold higher levels of CA activity as compared to untransformed controls and this activity was inhibited by CA-specific inhibitors. Sulfanilamide, acetazolamide, ethoxyzolamide, each at 10 mM, inhibited recombinant CA activity approximately 50%, 65%, and 75%, respectively. In plant tissue homogenates these inhibitors reduced CA activity by 50%, 70%, and 95%, respectively. Although CA activity was bighest in extracts of mature cotton leaves, probing total RNA with GhCA1 revealed CA transcript levels to be highest in the cotyledons of dark-grown cotton seedlings. Collectively, our data indicate the presence of a plastid localized CA in cotyledons of germinated seeds, suggesting a role for CA in postgerminative growth. PMID- 10682349 TI - Characterization of a flower-specific gene encoding a putative myrosinase binding protein in Arabidopsis thaliana. AB - A cDNA clone, 4B-1, previously isolated by differential screening is preferentially expressed in floral organs of Arabidopsis thaliana. Characterization of the full length cDNA and the genetic locus corresponding to 4B-1 cDNA revealed that it potentially encodes a myrosinase binding protein (MBP) which is presumably present in a large myrosinase complex. The deduced amino acid sequence of the polypeptide encoded by cDNA clone (designated f-AtMBP) appeared to consist of two parts: one region at the C-terminal half representing overall homology with AtMBP, an MBP homologue in A. thaliana, and the other at an extended N-terminal region of about 150 amino acids showing significant identity with the N-terminal region of the MBP-related protein reported in Brassica. Expression analysis by RNA blot and in situ hybridization showed that f-AtMBP was specifically expressed in floral meristems, pistils, stamens, petals, and ovules of immature flowers, but no expression was observed in the specialized cells called the myrosin cells in the hypocotyl and cotyledons of developing seeds where myrosinase enzymes are normally found. Although MBPs and MBP-related proteins are considered to be inducible by exogenous application of signal molecules and physical wounding, we found that f-AtMBP expression was not activated by such treatment, suggesting that f-AtMBP is a novel type of MBP specific to floral organs. PMID- 10682351 TI - Contaminants in the Greenland environment. PMID- 10682350 TI - Characterization of mitochondria-located small heat shock protein from tomato (Lycopersicon esculentum). AB - We cloned and sequenced a full-length cDNA encoding the precursor of the mitochondria-located small heat shock protein (MT-sHSP) gene (LeHSP23.8) from tomato (Lycopersicon esculentum). The deduced protein precursor with a calculated molecular weight of 23.8 kDa was predicted to target mitochondria and was classified as a plant MT-sHSP. A single copy of LeHSP23.8 was found in tomato genomic DNA by southern-blot analysis. Northern-blot analysis revealed the heat inducible character of LeHSP23.8 mRNA. The LeHSP23.8 mRNA was hardly detectable at about 36 degrees C but accumulated markedly at 40 degrees C. The molecular chaperone function of LeHSP23.8 was confirmed in vitro. The recombinant LeHSP23.8 was able to enhance the renaturation of chemically denatured citrate synthase (CS). Moreover, the recombinant LeHSP23.8 protected CS from thermal inactivation and also promoted the renaturation of thermally inactivated citrate synthase. PMID- 10682352 TI - Lead, cadmium, mercury and selenium in Greenland marine biota and sediments during AMAP phase 1. AB - Lead, cadmium, mercury and selenium levels in the Greenland marine environment from the first phase of the AMAP are presented. Samples were collected in 1994 1995 covering four widely separated regions in Greenland. Samples included sediments, soft tissue of blue mussel; and liver of polar cod, shorthorn sculpin, glaucous gull, Iceland gull and ringed seal. Concentrations of lead were found to increase with the size of blue mussel, but not with the age of gulls or ringed seal. Both cadmium and mercury concentrations were found to increase with the size/age of all species. Selenium concentrations decreased with increasing size of blue mussel, but increased with the age of gulls and ringed seal. Element levels found are within the range of those found in previous studies in Greenland. Relative to global background levels, lead levels must be considered low, whereas levels of cadmium, mercury and selenium in Greenland marine biota are high. Significant differences in element levels in sediments and biota among regions in Greenland were seen in several cases. There was a tendency for the highest lead and mercury concentrations to be found in east Greenland, whereas the highest cadmium concentrations were found in central west Greenland. However, the geographical differences among the media did not show a consistent pattern. PMID- 10682353 TI - An assessment of selenium to mercury in Greenland marine animals. AB - Information on mercury and selenium molar relation in muscle, liver and kidney tissue of Greenland marine animals is presented. In the majority of the samples selenium was present in a molar surplus to mercury. This was most clear in molluscs, crustaceans, fish and seabirds. A 1:1 molar ratio was found in tissues of marine mammals with high mercury concentrations (above approx. 10 nmol/g). This was most clearly demonstrated for liver and kidney tissue of polar bear and for ringed seal with high mercury concentration in the liver. These findings support previous results found in liver tissue of marine mammals, suggesting that methyl mercury is detoxified by a chemical mechanism involving selenium. If the anthropogenic release of mercury to the environment increases in the future due to increasing energy demands, species such as polar bears and seals with high tissue mercury concentrations should be monitored to elucidate whether this protective mechanism can be maintained in target organs. PMID- 10682354 TI - Geographical differences of zinc, cadmium, mercury and selenium in polar bears (Ursus maritimus) from Greenland. AB - Muscle, liver, and kidney tissues from 100 polar bears (Ursus maritimus), caught in the Avanersuaq area, north-west Greenland, and Ittoqqortoormiit area, central east Greenland, were analysed for zinc, cadmium, mercury and selenium. The zinc concentrations in muscle and liver were higher than in kidney. Mean zinc concentrations ranged from 19.7 to 76.0 micrograms/g (all data are presented as geometric means on a wet wt. basis). The presented cadmium concentrations by area and age groups were all low in muscle and in many cases below the detection limit (range: < 0.015-0.048 microgram/g). Cadmium concentrations were intermediate in liver (range: 0.120-1.98 micrograms/g) and highest in kidney tissue (range: 2.16 28.9 micrograms/g). Mercury was likewise lowest in muscle tissue (range: 0.034 0.191 microgram/g). Mercury concentration ranged quite similarly in liver and kidney tissue (liver range: 2.13-22.0 micrograms/g; kidney range: 2.87-32.0 micrograms/g). The selenium concentration increased from muscle (range: < 0.2 0.452 microgram/g) over liver (range: 1.20-9.80 micrograms/g) to kidney (range: 2.34-13.9 micrograms/g). No age accumulation was found for zinc. A weak increase was found for selenium, whereas cadmium and mercury clearly accumulated with age. An exception was mercury concentrations in muscle tissue, where no clear pattern was observed. Polar bears had significantly lower cadmium concentrations than ringed seals from the same area in all three tissues. Likewise mercury was significantly lower in the muscle tissue of polar bears than in ringed seals, whereas liver and kidney concentrations were higher. Biomagnification factors are provided for different tissues and age groups. Tissue ratios are given for different age groups and metals to enable a rough extrapolation from one tissue to another. Tissue ratios for cadmium, selenium and for mercury vary up to a factor of 6 with age. No significant differences could be detected between the elements analysed in bears from two management zones in north-west Greenland. This finding is in agreement with the genetic pattern in the two areas. In central-east Greenland, however, cadmium, selenium, and some of the mercury concentrations in polar bears from the southern area were higher than from the northern area, indicating that the east Greenland area represents two different ecological regions with different polar bear populations. Geographical differences between polar bears from north-west and east Greenland were only found for mercury and cadmium in liver tissue, where the concentrations were highest in bears from north-west Greenland. The geographical trend of increasing cadmium concentrations in polar bear liver tissue from west to east, which has been found previously in Canada, could be extended to cover north-west Greenland as well. East of this region a decrease was found. Mercury concentrations in polar bear liver tissue showed an increase from Svalbard over east and north-west Greenland, peaking in bears from south-west Melville Island. A marked decrease was found west of Melville Island, and the lowest concentrations were found in the Chukchi Sea. PMID- 10682355 TI - Temporal trends of cadmium and mercury in Greenland marine biota. AB - Data for cadmium and mercury in Greenland marine biota (blue mussels, polar cod, shorthorn sculpin, glaucous gull and ringed seals) over a period of 20 years has been analysed in order to assess temporal changes. Most of the comparisons were conducted between tissue samples collected in the mid-1980s and mid-1990s. Cadmium data from a few time series obtained at reference sites during monitoring of mining activities were also included. No overall temporal trends in cadmium or mercury concentrations were found within the 20-year period assessed. However, cadmium concentrations in ringed seals tended to increase in the period from late 1970s to the mid-1980s. From the mid-1980s to the mid-1990s cadmium concentrations in ringed seals decreased again, whilst mercury concentrations showed a tendency to increase in the same period. The observed changes may reflect natural fluctuations caused by factors such as a shift in feeding behaviour, rather than changes in anthropogenic exposure. PMID- 10682356 TI - Mercury in dated Greenland marine sediments. AB - Twenty marine sediment cores from Greenland were analyzed for mercury, and dated by the lead-210 method. In general the cores exhibit a mercury profile with higher mercury concentrations in the upper centimetres of the core. The cores were studied by linear regression of in Hg vs. age of the sediment for the youngest 100 years. As a rule the mercury decreased with depth in the sediment with various degrees of significance. The increase of the mercury flux during the last 100 years is roughly a doubling. The increase may be of anthropogenic origin as it is restricted to the last 100 years. In four cores the concentration of manganese was found also to increase in the top layers indicating diagenesis. In the other cases the higher concentrations were not accompanied by higher manganese concentrations. The mercury flux to the sediment surface was generally proportional to the Pb-210 flux indicating that the mercury mainly originates from atmospheric washout. But the large variability indicates that other processes also influence the mercury flux to Arctic marine sediments. PMID- 10682357 TI - Temporal and spatial trends of persistent organochlorines in Greenland walrus (Odobenus rosmarus rosmarus). AB - Persistent organochlorines [PCBs, DDT and chlordane related compounds, dieldrin, toxaphene, hexachlorocyclohexane (HCH), chlorobenzenes] were determined in blubber of Atlantic walrus (Odobenus rosmarus rosmarus) in 1978 and 1988 from the Avanersuaq (Thule) region of north-west Greenland and in 1989 from Ittoqqortoormiit (Scoresbysund) in east Greenland. Lowest concentrations of organochlorines (OCs) were found in the samples from the Avanersuaq region while much higher levels of all compounds, except HCH isomers and mono/dichlorobiphenyls (CB5/8), were observed in samples (all males) from Ittoqqortoormiit. Total PCBs (sigma PCB) averaged 246 ng/g (wet wt.) male walrus from Avanersuaq and 2860 ng/g in samples from Ittoqqortoormiit. DDT isomers showed the greatest difference between the two locations, 50 x for p,p'-DDE and 69 x higher for p,p'-DDT. Ittoqqortoormiit walrus showed the pattern of OCs characteristic of seal-eating animals although the consumption of other organisms cannot be ruled out. The higher levels of OCs in east Greenland compared to north west Greenland animals were consistent with results for polar bears, seals and gulls from the same regions. Principal components analysis showed that the pattern of OCs in Ittoqqortoormiit walrus was very similar to that in walrus from Inukjuaq in east Hudson Bay, which have previously been reported to be seal eaters, and quite distinct from the Avanersuaq walrus. No significant differences in mean concentrations of any OCs were found between male walrus from 1978 and 1988. For females, there were significantly higher levels of CB5/8, trichlorobiphenyls, dieldrin, toxaphene and alpha HCH as well as sigma HCH but not for sigma PCBs or DDT compounds. The data for Greenland walrus from the 1970s and late 1980s provide a baseline for future trend monitoring in walrus. PMID- 10682358 TI - Organochlorines in Greenland marine fish, mussels and sediments. AB - Shorthorn sculpin (Myoxocephalus scorpius), polar cod (Boreogadus saida), blue mussels (Mytilus edulis) and sediments were sampled in Greenland 1994-1995 at three locations at the west coast and one at the east coast. Fish liver, mussel soft tissue and sediments were analysed for PCBs (10 congeners), DDTs (pp,'), HCHs (alpha, beta, gamma), HCB and trans-nonachlor. The overall geometric mean concentrations found for PCBs were 17 micrograms kg-1 wet wt. in shorthorn sculpin liver, 33 micrograms kg-1 wet wt. in polar cod liver, and 0.86 microgram kg-1 wet wt. in blue mussels. For the three species, the geometric mean concentrations for DDTs were 11, 36, and 0.39 micrograms kg-1 wet wt., respectively; for HCHs: 8.7, 32 and 0.56 micrograms kg-1 wet wt., respectively; for HCB: 4.2, 11 and 0.06 micrograms kg-1 wet wt., respectively; and for trans nonachlor: 6.3, 19 and 0.16 microgram kg-1 wet wt., respectively. All organochlorines in the sediment samples were below the detection limit of 0.1 microgram kg-1 dry weight. For sculpins and mussels, most organochlorine compounds were found to increase with increasing lipid content. The weight of mussels did not influence organochlorine concentrations, whereas organochlorine content in general increased with fish length of sculpins. The concentrations were found to be comparable to levels in other Arctic regions, but orders of magnitude lower than levels found in the southern part of the North Sea. Organochlorine concentrations in sculpins showed a decreasing trend following the ocean current flowing from north to south at the east coast and from south towards north at the west coast of Greenland. The proportion of higher chlorinated PCBs (Cl atoms > or = 6) in sculpin liver followed the decreasing trend of PCB concentrations. PMID- 10682359 TI - Organochlorines in Greenland ringed seals (Phoca hispida). AB - Twenty-five ringed seals (Phoca hispida) were sampled in 1994 at each of four areas in Greenland, three on the west coast and one on the east coast. Seal blubber samples were analysed for polychlorinated biphenyls (PCBs, IUPAC Nos. 28, 31, 52, 101, 105, 118, 138, 153, 156 and 180), DDTs (p,p'-DDE, p,p'-DDD, p,p' DDT), hexachlorocyclohexanes (alpha-, beta- and gamma-HCH), hexachlorobenzene (HCB) and trans-nonachlor. A number of the seals (n = 56) were also analysed for toxaphene (total toxaphene and single congeners: CHB Nos. 26, 32, 50 and 62). The overall geometric means (and ranges) found in seals were in microgram kg-1 wet wt. for sigma PCBs: 452 (85-4200), sigma DDTs: 607 (97-6040), sigma HCHs: 123 (51 382), HCB: 13 (3-27), trans-nonachlor: 83 (21-1236) and total toxaphene: 263 (71 950). The geometric means for HCB, trans-nonachlor, toxaphenes and the sums of PCBs and DDTs were higher for the east coast samples than for those from the west coast, and a decreasing trend in concentrations followed the east Greenland current. No geographical trend was apparent for HCHs. Differences in concentrations between females and males were only significant for HCB and HCH within certain age classes and sampling areas. A tendency for concentrations to increase with age was observed, but was not statistically significant. Principal component analyses for PCB congeners revealed that the proportion of higher chlorinated PCBs (Cl atoms > or = 6) decreased with decreasing concentrations of PCBs found for the four sampling areas. No differences between the higher and lower chlorinated PCBs were seen in females and males, but a higher proportion of CB-105 and CB-118 compared to CB-52 and CB-101 was seen in male seals compared to female seals. PMID- 10682361 TI - TBT in marine sediments and blue mussels (Mytilus edulis) from central-west Greenland. AB - Concentrations of butyltin compounds were investigated in the bivalve Mytilus edulis (five sites) and marine sediments (three sites) near the largest town, Nuuk, in Greenland. In seven of the eight samples the extremely toxic compound tributyltin (TBT) was detected. The concentrations of tributyltin and degradation products in the bivalves were close to 1 microgram kg-1 wet weight (ww), calculated as Sn, which is lower than those found in Iceland and the Faeroe Islands. In sediments the concentration of TBT ranged from below the limit of detection of 1 microgram kg-1 to 171 micrograms kg-1 dry weight (dw), calculated as Sn, which is comparable to levels found in Europe. PMID- 10682360 TI - Organochlorines in Greenland glaucous gulls (Larus hyperboreus) and Icelandic gulls (Larus glaucoides). AB - Glaucous gulls (Larus hyperboreus) and Icelandic gulls (Larus glaucoides) were sampled in 1994 from four different areas in Greenland, three on the west coast and one on the east coast. Livers of 93 glaucous gulls and seven Icelandic gulls were analysed for polychlorinated biphenyls (PCBs, IUPAC Nos. 28, 31, 52, 101, 105, 118, 138, 153, 156 and 180), DDTs (p,p'-DDE, p,p'-DDD, p,p'-DDT), hexachlorocyclohexanes (alpha-, beta- and gamma-HCH), hexachlorobenzene (HCB) and trans-nonachlor (TNC). The overall geometric means of the concentrations found in glaucous gull liver were for sigma PCBs 388 (range 20-5557), for sigma DDTs 363 (17-8604), sigma HCHs 7.4 (1-53), HCB 47 (4-594) and trans-nonachlor 19 (3-187) micrograms kg-1 wet wt., respectively. The geometric means of concentrations in Icelandic gull liver were for sigma PCBs 112 (24-435), for sigma DDTs 95 (25 298), sigma HCHs 2.9 (1.4-5.2), HCB 22 (8-58) and trans-nonachlor 5.1 (2.4-8.6) micrograms kg-1 wet wt., respectively. Significantly (P = 0.05) higher concentrations of PCBs, DDTs and HCHs were found in glaucous gulls at Ittoqqortoormiit at the east coast than in gulls from Qeqertarsuaq at the west coast of Greenland. This tendency was also seen for HCB and trans-nonachlor, but the differences were not statistically significant (P = 0.05). A decreasing trend in organochlorine concentrations followed the East Greenland Current, flowing from north to south down the east coast and to the north on the west coast. Gulls taken from the most northerly sampling area of the west coast, however, showed slightly higher concentrations than those from the central west coast. There appeared to be a tendency for higher concentrations to be found in males than females, and in adults compared to young glaucous gulls, but the differences were not statistically significant (P = 0.05). The concentration ranges found in gulls from Greenland were similar to those reported previously for gulls from northern Norway and Russia. A principal component analysis revealed no obvious link between the presence of higher chlorinated PCBs and higher PCB concentrations in glaucous gulls. Significantly higher proportions of higher chlorinated PCBs were found in glaucous gulls than in Icelandic gulls, and in adult glaucous gulls compared to young gulls of 1-2 calendar years. As no such difference was found between female and male gulls it seems that PCBs of all degrees of chlorination may be passed equally well from mother to offspring. PMID- 10682362 TI - The use of lichen (Cetraria nivalis) and moss (Rhacomitrium lanuginosum) as monitors for atmospheric deposition in Greenland. AB - Concentrations of Pb, Cd, Hg, Zn, Cu, Cr, Ni, As, V, Al and Fe are reported from soil, humus, moss (Rhacomitrium lanuginosum) and lichen (Cetraria nivalis) sampled at four locations in Greenland. For Al, Fe, Cr and V the levels in soil were highest followed by humus and R. lanuginosum and with the lowest levels in C. nivalis. The same was true for Pb, Cu and Ni but without as great a difference between medias. For Cd and Hg, the lowest levels were found in soil. For Zn and As, the media with highest levels differed between locality. Data were examined by a principal component analysis. Three principal components explained 87% of the total variation. The dominant elements in the first component were Fe, Al, V, Ni, Cr, Cu and Pb. This component is interpreted as a soil dust factor. The concentrations in R. lanuginosum and C. nivalis of these elements are believed to be highly influenced by soil dust. Pb concentrations in moss and lichen may also be influenced by other sources as Pb also had some correlation's with the third component. Zn and Cd and to a lesser extent. As were the dominant elements in the second component. The third component was highly dominated by Hg with a lesser influence of Pb and As, Zn, Cd and Hg concentrations in R. lanuginosum and C. nivalis are believed to be influenced by other sources than soil dust which may be long-range atmospheric transport. In general, both the within locality and the between locality variability in the values of the three components decreased in the order soil, humus, R. lanuginosum and C. nivalis. The lichen C. nivalis is looked at as an indicator with greater potential for monitoring atmospheric deposition of elements than the moss R. lanuginosum. PMID- 10682363 TI - Lead, zinc, cadmium, mercury, selenium and copper in Greenland caribou and reindeer (Rangifer tarandus). AB - Samples of caribou and reindeer muscle (127 samples) and liver (126 samples) were collected from four locations during two seasons plus 3 years in Greenland. The levels of lead, zinc, cadmium, mercury, selenium, and copper were determined, and analyzed in relation to location, two seasons, age and year of sampling. The lead concentrations (geometric mean) ranged from below the detection limit to 0.007 microgram/g wet weight (wet wt.) in muscle and from 0.027 to 0.926 microgram/g wet wt. in liver. Zinc geometric mean concentrations ranged from 17.5 to 39.6 micrograms/g wet wt. in muscle and from 23.2 to 31.7 micrograms/g wet wt. in liver. For cadmium, the geometric mean concentrations were at, or below the detection limit in muscle, while concentrations in liver ranged from 0.121 to 0.695 microgram/g wet wt. Mercury levels ranged from 0.003 to 0.043 microgram/g wet wt. in muscle and from 0.040 to 0.618 microgram/g wet wt. in liver. Selenium concentration levels in muscle ranged from 0.030 to 0.252 microgram/g wet wt., and from 0.085 to 0.984 microgram/g wet wt. in liver. Copper levels in muscle ranged from 2.09 to 3.60 micrograms/g wet wt., and from 21.8 to 71.0 micrograms/g wet wt. in liver. Mercury concentrations were higher than those found at lower latitudes in Norway and Canada, especially in Isortoq in southern Greenland. Selenium levels were also high compared to other Arctic regions. Concentrations of lead, zinc, cadmium and copper are similar to those reported in caribou from Canada and Norway. Concentrations of elements generally decreased in the following order: Isortoq > Akia > Itinnera > Kangerlussuaq, and there was only found minor variation in the annual levels during 3 years in Itinnera. Late winter levels were generally significantly higher than early winter levels especially in the lichen-rich localities, and it is suggested that the availability of lichens as winter forage is the key determining the level of elements. Accordingly, when using caribou and reindeer as monitoring organism, knowledge of winter forage is very important for interpretation of results. PMID- 10682364 TI - Mercury in Arctic char (Salvelinus alpinus) populations from Greenland. AB - Mercury concentrations were determined in muscle tissue of lake resident and anadromous populations of Arctic char in Greenland. Mercury in lake sediment, and in soil and humus from the surrounding area were also determined in the main localities. Fish length and dry weight were shown to be important covariables, which have to be taken into account when comparing mercury levels between populations. Variations in fat content did not contribute further to the differing mercury concentrations. Mercury concentrations in lake sediments, humus from around the lakes and resident populations of Arctic char from west Greenland and south-west Greenland were higher than for populations from east Greenland and north-west Greenland. The mercury level in anadromous populations was found to be 10-15-fold lower than that found in lake resident populations, and similar to that found in marine fish species. Methyl mercury was determined in two of the populations investigated, and constituted 72-92% of the total mercury. PMID- 10682365 TI - Organochlorines in Greenland lake sediments and landlocked Arctic char (Salvelinus alpinus). AB - Lake sediments and landlocked Arctic char (Salvelinus alpinus) were sampled in 1994 and 1995 at four different locations in Greenland, three at the west coast and one at the east coast. Sediments, char muscle and char liver were analysed for PCBs (10 congeners), DDTs (pp'), HCHs (alpha, beta, gamma), dieldrin, HCB and chlordanes (5) and char in addition to toxaphene (total and 4 congeners). All organochlorines in the sediment samples were below the detection limit of 0.1 microgram kg-1 dry wt., while the overall geometric means in Arctic char muscle were, in microgram kg-1 wet wt., for PCBs 11 (range 1-140), for DDTs 4.0 (1-35), for HCHs 0.4 (0.06-1.5), for dieldrin 0.7 (< 0.1-4.2), for HCB 0.7 (0.09-3.8), for chlordanes 4.8 (1-57) and for total toxaphene 13 (1-180). The sums of PCBs, DDTs, chlordanes and total toxaphene disclosed higher concentrations in muscle of char from the east coast compared to char from the west coast. Dieldrin and HCB showed the same tendency, but less pronounced, while the sum of HCHs were found in highest concentrations at the west coast. PCB, DDT, chlordane and toxaphene concentrations showed a decreasing trend following the East Greenland Current. Principal component analysis on PCB congeners showed that the proportion of higher chlorinated PCBs (Cl > 5) were higher in Qaqortoq in south Greenland compared to the three other sampling areas in Greenland. However, no correlation was seen either between degree of chlorination and latitude or between degree of chlorination and PCB concentrations. PMID- 10682366 TI - Lead, cadmium, mercury and selenium intake by Greenlanders from local marine food. AB - The human intake of lead, cadmium, mercury and selenium from local Greenlandic marine food was estimated based on dietary studies and contaminant information. The average lead intake was calculated to be 15 micrograms/person per week, which is very low, whereas the intake of cadmium and mercury was estimated to be very high, on average 1004 micrograms/person per week for cadmium and 846 for mercury, thereby significantly exceeding limits established by FAO/WHO. The main cadmium and mercury source was seal liver. Selenium intake was also high with whale skin as the dominant source. PMID- 10682367 TI - Organochlorines and heavy metals in pregnant women from the Disko Bay area in Greenland. AB - Recent studies from Greenland and the Canadian Arctic have shown high concentrations of heavy metals, such as mercury, and organochlorines in the blood and fatty tissue of the Inuit. This is attributed in particular to their high consumption of the meat and blubber of marine mammals. In the present study, 180 pregnant women and 178 newborn babies were studied, amounting to 36% of the total number of births in the Disko Bay area during 1994-1996. The pesticides found in the highest concentrations in maternal blood were DDE (4.8 micrograms/l wet wt.), trans-nonachlor (1.6 micrograms/l) and hexachlorobenzene (1.2 micrograms/l) while the total concentration of PCB (Aroclor 1260) was 19.1 micrograms/l. Calculated on a lipid basis, concentrations were slightly higher in maternal than in cord blood. The mercury concentrations were 16.8 micrograms/l in maternal blood and 35.6 micrograms/l in cord blood. In a linear regression analysis, the concentrations of organochlorines, mercury and selenium increased with maternal age. Concentrations of mercury and cadmium increased with the consumption of marine mammals, and cadmium was associated with smoking. The contaminants are potentially toxic for several organ systems but the high concentrations of pollutants have so far not been shown to influence health in Greenland. PMID- 10682368 TI - Analytical methods, quality assurance and quality control used in the Greenland AMAP programme. AB - The majority of analytical results in the Greenland AMAP (Arctic Monitoring and Assessment Programme) have been produced by laboratories that participate regularly in performance studies. This makes it possible to judge the quality of the results based on objective measurements made by independent assessors. AMAP laboratories participated while analysing the AMAP samples in the QUASIMEME laboratory performance study programme, in the 'Interlaboratory Comparison Program' organised by Le Centre de Toxicologie du Quebec, in a toxaphene intercomparison study organised by The Food Research Division of Health Canada, and in an International Atomic Energy Agency Intercomparison exercise. The relative errors of the trace analyses, i.e. the relative deviation of the result obtained by the AMAP laboratory from the assigned value, are in most cases less than the 25% which is regarded as acceptable by QUASIMEME. Usually the errors, especially for trace elements, are less than 12.5%, while errors for trace organics below 1 microgram kg-1 may rise to 50% or more. This study covers the period 1993 to 1998 for trace elements and one or more years from the period 1994 1996 for trace organics. PMID- 10682369 TI - Comparison of contaminants from different trophic levels and ecosystems. AB - The present paper provides an overview of the priority contaminants and media from the Greenland part of the Arctic Monitoring and Assessment Program. Levels and accumulation patterns of heavy metals, POPs and a radionuclide (137Cs) are compared from the terrestrial, freshwater and marine ecosystems. Of the nine compounds presented, seven (Cd, Hg, Se, sigma PCB, sigma DDT, sigma HCH, HCB) increased in concentration towards higher trophic levels. For these contaminants the concentrations in soil and aquatic sediment were in the same order of magnitude, whereas the concentrations in marine biota were higher than found in the freshwater and terrestrial ecosystems probably due to the presence of longer food chains. Pb and 137Cs showed the reverse pattern compared with the other compounds. The concentrations in soil and aquatic sediments decreased in the order terrestrial, freshwater and marine ecosystems, which was reflected in the biota as well. Reindeer had similar or lower levels of Pb and 137Cs than lichens. Levels of Pb and 137Cs in marine biota did not show the same clear increase towards higher trophic as found for the other analysed compounds. Greenland Inuit contains considerably less mercury but higher levels of sigma PCB, sigma DDT and HCB than other Arctic marine top consumers. PMID- 10682370 TI - Environmental radioactive contamination in Greenland: a 35 years retrospect. AB - Environmental studies of anthropogenic radionuclides in Greenland over four decades are reported. The studies have comprised the marine as well as the terrestrial environments and emphasis has been laid on measurements of 90Sr and 137Cs. The temporal and the spatial trends of these radionuclides are described. The radiation exposure from consumption of locally produced diets has been calculated from consumption rates and the infinite time integrated levels of 90Sr and 137Cs concentrations in the various food products. Compared with most other Arctic people, the Greenlanders have received relatively low doses from anthropogenic radionuclides. There are several reasons for this, first of all, because of the relatively high consumption of marine products compared with terrestrial products. Secondly, because winter slaughtering of reindeer is less frequent in Greenland than in other Arctic countries and Greenland reindeer consume, in general, less lichen than most other Arctic reinder, and thirdly, because the transfer from deposition to lichen in Greenland seems lower than in other Arctic areas. PMID- 10682371 TI - Evaluation of the Greenland AMAP programme 1994-1995, by use of power analysis (illustrated by selected heavy metals and POPs). AB - The levels of PCBs, HCB, HCHs, DDTs, Cd, Pb, Hg and Se, and especially the variability in biota obtained during Phase 1 of the Greenland AMAP-programme have been used to illustrate the ability of the programme to detect differences in contaminant levels over time. The statistical power of t-tests of contaminant levels are illustrated according to various scenarios of magnitude of change, significance level and sample size. The statistical power of various time series of contaminant levels to detect linear trends in mean log-concentrations, including a random between-year variation component, is illustrated. We conclude that the ability to detect differences is rather poor for many combinations of contaminants and media, and that long time series are needed before temporal trends are likely to be detected. PMID- 10682372 TI - Critical loads of acidity for surface waters in China. AB - For further control of acid rain and sulphur dioxide pollution, the Chinese government has designated the Acid Rain Control Zone and the Sulphur Dioxide Pollution Control Zone for those areas that are, or could become, affected by acid deposition or ambient sulphur dioxide concentrations. One of the most important principles for designating the Acid Rain Control Zone is that the critical load is exceeded by the sulphur deposition. Through the steady-state water chemistry method (SSWC), critical loads of acidity for surface waters were mapped based on available data. Results show that surface waters sensitive to acid deposition, i.e. surface waters with low critical loads, are mainly found in north-east China, on the Tibetan Plateau, and in north-west China. Compared with the critical loads of soils, critical loads of surface waters are usually higher in almost all areas in China. The reason for very low critical loads of surface waters in some regions dominated by soils geologically not sensitive to acid deposition may be attributed to the low temperature, high altitude and low runoff. In contrast, surface waters in south China are not susceptible to acid deposition, and so far acidification of surface water has not been found in spite of the heavy acid rain. As can be seen from the critical load exceedance map, nearly 10% of the surface waters are subject to risk of acidification in 1995. PMID- 10682373 TI - Quantitative structure-property relationships for direct photolysis quantum yields of selected polycyclic aromatic hydrocarbons AB - By the use of partial least squares (PLS) method and 11 quantum chemical descriptors computed by PM3 Hamiltonian, Quantitative Structure-Property Relationships (QSPRs) for direct photolysis quantum yields of selected polycyclic aromatic hydrocarbons (PAHs) were obtained. Direct photolysis quantum yields were predicted for PAHs for which experimental quantum yield values were lacking. Based on the QSPR models, significant PAH molecular characters governing their direct photolysis quantum yields were identified. It can generally be concluded that PAHs with large average molecular polarizability, molecular weight, and heat of formation values tend to have small photolysis quantum yields. PAHs with large values of the energy of the lowest unoccupied molecular orbital (Elumo), small values of the energy of the highest occupied molecular orbital (Ehomo), and large Elumo-Ehomo values, tend to have great photolysis quantum yields. PMID- 10682374 TI - The chemistry of Norwegian groundwaters: III. The distribution of trace elements in 476 crystalline bedrock groundwaters, as analysed by ICP-MS techniques AB - Four hundred and seventy-six groundwater samples from boreholes in Norwegian crystalline bedrock have been analysed by ICP-MS techniques. The results for 53 trace elements are presented as cumulative frequency distribution diagrams and are compared with relevant international drinking water norms. A range of trace elements appear to be enriched in granitic waters and depleted in anorthositic waters which is to be expected as generally granitic rocks are enriched in trace elements above those in anorthosites. A selection of elements which may be toxic in excess when present in drinking water are further discussed (Be, Tl, Th, U, Cd, Pb, As, Ni, and Hg). For uranium, 18% of the samples exceed the American maximum admissible concentration of 20 micrograms/l; 7% of the samples fail to meet the Russian drinking water norms for beryllium of 0.2 microgram/l. For some parameters such as U, Be and Tl, no Norwegian drinking water regulations are set, while the American and the Russian norms differ significantly from each other. Between 0 and 1.5% of the wells exceed Norwegian drinking water norms for each of the other selected elements. PMID- 10682375 TI - Inhalation exposure to THMs from drinking water in south Taiwan. AB - Trihalomethanes (THMs) are important disinfection byproducts (DBPs) in drinking water. To understand the magnitude of exposure to THMs for the people in southern Taiwan, models are used to estimate the inhalation exposure associated with drinking water based on raw water quality. Two parts of models are used in this study, one for estimating THM concentration from raw water quality, and one for estimating inhalation exposure to people. Important raw water quality and operational parameters, including TOC, UV254, pH, temperature, chlorine dosage, and water residence time of a major water treatment plant in south Taiwan were collected. An empirical THM formation model was then employed to predict the THM concentration at consumers' dwellings based on the parameters collected. Differences between the predicted results and experimental data were found to be small, indicating that the model is appropriate. The predicted THM concentration distribution was served as input parameters for the exposure models. Three major scenarios associated with probable inhalation exposure of THMs, including shower, pre- and post-cooking activities, and cooking processes, were considered in the exposure models. The model results show that the mean inhalation exposure of THMs for shower, pre- and post-cooking activities, and cooking processes are 26.4, 1.56, 3.29 micrograms/day, respectively. The total inhalation exposure (summation of the three scenarios) was found to be comparable with that for direct ingestion, indicating that inhalation is an important pathway for THM exposure from drinking water. PMID- 10682376 TI - The influence of a capacitor plant in Serpukhov on vegetable contamination by polychlorinated biphenyls. AB - PCB content in soil and vegetables grown on the polluted soils in some districts of the town of Serpukhov have been studied for 10 years after the use of PCBs had been banned at the local capacitor plant. Soil contamination with PCBs in the vicinity of the plant is still extremely high (up to 30 mg/kg). Vegetables grown on the polluted soils, especially carrots and green parts of fennel, parsley, celery are also highly contaminated. The primary pollutants are found to be tri- and tetrachlorobiphenyls (up to 70-80% of total PCBs). PMID- 10682377 TI - Concentrations of mercury, cadmium, lead and copper in fruiting bodies of edible mushrooms in an emission area of a copper smelter and a mercury smelter. AB - Four metals were determined by AAS techniques in 56 samples of 23 wild mushroom species collected in a heavily polluted area in eastern Slovakia in 1997 and 1998. The area has been contaminated from historical polymetallic ores mining and smelting and by emissions from a mercury smelter between 1969 and 1993 and from a copper smelter since 1951. No significant differences in metal concentrations (P < 0.05) were found in four species when comparing the periods 1992-1993 and 1997 1998. Considerable contamination of most species was observed mainly for mercury and cadmium. The highest levels of mercury, up to 50 mg kg-1 dry matter, were found in Boletus reticulatus, Lycoperdon perlatum and Marasmius oreades, and of cadmium up to 20 mg kg-1 dry matter in Xerocomus chrysenteron and Lycoperdon perlatum. The latter species also had extremely high lead and copper concentrations in hundreds of milligrams per kilogram dry matter. Concentrations of mercury and copper in caps of four Boletaceae species were significantly (P < 0.05) higher than those in stipes. PMID- 10682378 TI - A biokinetic model for lead metabolism with a view to its extension to pregnancy and lactation; (1). Further validation of the original model for non-pregnant adults. AB - A published biokinetic model that describes the absorption, transfer between organs and excretion of lead (Pb) in humans has been established using commercially available simulation software. Recent in vivo data have been used to validate further the model in adults, particularly for non-pregnant females. The validation data centre on: (a) the prediction of blood Pb concentrations due to changes in atmospheric and dietary Pb levels over the last 25 years; (b) the non linear relationship between Pb in whole blood and that in blood plasma which can be transferred to other organs; and (c) the accumulation of Pb in bone which may be re-mobilised later in time of calcium stress. This work underpins our alteration of the model to encompass pregnancy and lactation so that the build-up of Pb in the developing foetus and breast-fed infant can be estimated from any number of current and historical maternal exposure scenarios. PMID- 10682380 TI - Trends in veterinary clinical and fundamental pharmacology: past and future in The Netherlands. AB - Veterinary pharmacology has undergone a gradual development in the Netherlands during this century. Starting from a historical perspective the paper aims to provide an overview of future trends and important issues in the area of veterinary pharmacology and toxicology. It is pointed out that this discipline comprises several subdisciplines as the comparative aspect of both, pharmacology and toxicology, is inherent to veterinary medicine which has to address a broad variety of animal species. Thus, the comparison of drug effects, side effects, and drug disposition as well as the comparison of the species-specific susceptibility to xenobiotics are obvious challenges in this discipline. Several areas in clinical pharmacology are highlighted to indicate future research needs. Finally, the principles of Good Veterinary Practice are presented as the 'golden standard' in veterinary clinical pharmacology. PMID- 10682379 TI - Response of stress indicators and growth parameters of Tibouchina pulchra Cogn. exposed to air and soil pollution near the industrial complex of Cubatao, Brazil. AB - The present study was performed in the vicinity of the industrial complex of Cubatao, Sao Paulo, Brazil, in order to evaluate the response of 'manaca da serra' Tibouchina pulchra Cogn. (Melastomataceae), a common species of secondary Atlantic Rain Forest vegetation, to the impact of complex air pollution. Emphasis was given to changes of biochemical parameters such as ascorbic acid concentration, peroxidase activity, contents of water-soluble thiols, pH of leaf extract and buffering capacity. These plant factors are often used as early indicators of air pollution stress. Field experiments included sampling of leaves from mature trees in areas with different air pollution load (passive monitoring), exposure of saplings cultivated in uniform soil at these areas (active monitoring) and a study on the combined effects of contaminated soil and air pollution. In general, metabolic response of saplings was more accentuated than that of mature trees. Leaf extract pH and buffering capacity showed no or only small alterations in plants exposed to industrial emissions. In contrast, air pollution resulted in a distinct decrease in ascorbic acid contents and an increase in peroxidase activity and thiol concentrations in leaves. Cultivation of saplings in soil types from contaminated regions frequently caused the same modifications or enhanced the effects produced by air pollution. Growth analysis of exposed saplings demonstrated that a change of the relationship between above ground and below-ground plant parts was the most obvious effect of air pollution and soil contamination. The experiments showed that even T. pulchra, a species considered resistant to air pollution, suffers metabolic disturbances by the present ambient air and soil quality. Although biochemical and physiological alterations were not related to a certain air pollution type, they could be used to estimate the overall pollution load and to map zones with different air quality. PMID- 10682381 TI - Signal transduction in inflammatory processes, current and future therapeutic targets: a mini review. AB - The selective control of inflammatory reactions will continue to be a major issue in the development of new drugs. Many new molecular targets are coming up. This paper highlights a few key mediators that are nowadays considered as interesting therapeutic intervention points. Cytokines play an important regulatory role in the initiation, maintenance and termination of inflammatory reactions. More than 50 cytokines have been identified, and more and more has become known about their receptors and signal transduction pathways. Tumour necrosis factor-alpha (TNF alpha) is still regarded as one of the initial cytokines of the cascade, and different approaches are followed to control its synthesis, release or effects. Lipopolysaccharide (LPS) is a one of the triggers that is able to induce a strong TNF-response. Inhibitors of cyclic nucleotide phosphodiesterases (PDEs), including rolipram and pentoxifylline suppress the LPS-induced TNF-alpha production in monocytes/macrophages. In our laboratory it has been shown that the alternative way to increase cAMP levels, via stimulation of beta-adrenergic receptors, also provides an effective way, both in vitro and in vivo, to inhibit TNF-alpha release. Other therapeutic ways include the use of antibodies directed to cytokines, TNF receptor fused to IgG, antibody therapy against TNF, the use of MAP kinase inhibitors. The different signal transduction pathways, including the NF-kappa B activation route may provide alternative pharmacological tools. We may surely expect anti-inflammatory drugs of much greater specificity to be developed in the next decade. Despite the relative limited investments in veterinary drug development this will also have consequences for veterinary therapy. PMID- 10682382 TI - Cytochromes and cytokines: changes in drug disposition in animals during an acute phase response: a mini-review. AB - Diseases are a major cause of variation in drug response. Although many different diseases are known that have an effect on the pharmacokinetics or sometimes the pharmacodynamics of a drug, disorders associated with a so-called acute phase response (APR) are the most important in this respect. During APR, for example caused by tissue damage or invasion of a pathogen, a group of symptoms can be observed that often include fever, lassitude, inhibition of gastric function and synthesis of acute phase proteins. All phases that together determine the pharmacokinetic profile of a drug, absorption, distribution, metabolism and excretion, can be affected during APR. From a clinical point of view however, the effects on absorption and metabolism are the most relevant. For drugs that are given orally, a slower absorption rate is often observed during APR due to a delayed gastric emptying. Even more important from a clinical point of view is the depression of biotransformation capacity in the liver during APR, especially affecting the enzymes of the cytochrome P450 (CYP450) complex. Although much has become known about the mechanism of this effect, a number of questions remain. Cytokines, nitric oxide and possibly the enzyme heme oxygenase are playing a role in a complex process that depends on a mutual interaction between Kupffer cells (macrophages) and hepatocytes in the liver. The clinician should be aware of unexpected changes in drug effects or residue levels due to cumulation of the compound during disease or after vaccination. In these situations, drugs that are excreted unchanged may be better alternatives. PMID- 10682383 TI - Cocultures of porcine hepatocytes and Kupffer cells as an improved in vitro model for the study of hepatotoxic compounds. AB - In this study primary hepatocyte cultures (HC cultures) and cocultures comprised of hepatocytes and Kupffer cells (HC/KC cocultures) were compared to investigate the inflammatory response induced by lipopolysaccharide (LPS). In addition both culture types were compared to study the hepatotoxic effects of two frequently used drugs: tiamulin and chlorpromazine. The inflammatory response in both culture types was determined by measurement of tumour necrosis factor-alpha (TNF alpha), interleukin 6 (IL-6) and nitric oxide (NO). The drug-induced hepatotoxic effects were determined by measuring production of intracellular reactive oxygen species (ROS) and cytotoxicity. Exposure of both cultures to LPS resulted in a significantly increased production of TNF-alpha, IL-6 and NO. However, the production of TNF-alpha, IL-6 and NO was substantially increased in culture supernatant of cocultures, compared to single HC-cultures. Both tiamulin and chlorpromazine were potent inducers of intracellular ROS production at concentrations > or = 50 microM. High ROS production was paralleled by increased cytotoxicity as observed in both culture types. Incubation of cocultures with chlorpromazine resulted in a significant increased ROS production as compared to HC cultures. In contrast, no significant differences between HC-cultures and HC/KC cocultures were observed for tiamulin induced ROS production or cytotoxicity. The present study demonstrates that cocultures between Kupffer cells and hepatocytes provide an excellent model for the study of hepatotoxic compounds which exert (part) of their toxic effects via the activation of Kupffer cells. Furthermore they offer a valuable tool to study increased susceptibility to intoxication from xenobiotic agents in case of a concurrent or pre-existing inflammation. PMID- 10682384 TI - Suppression of the acute inflammatory response of porcine alveolar- and liver macrophages. AB - During infection and inflammation drug disposition and hepatic metabolism are markedly affected in mammals. Pro-inflammatory mediators play an important role in the suppression of (cytochrome-P450-mediated) drug metabolism. Inflammatory mediators like cytokines, nitric oxide (NO), reactive oxygen species (ROS) and eicosanoids are released by activated macrophages from various sources, including liver and lung. It was the aim of this study to investigate ways to suppress the activation of macrophages during the onset of the inflammatory cascade. Therefore porcine lung and liver macrophages were isolated, and incubated with lipopolysaccharide (LPS) to initiate an acute inflammatory response, represented by the release of high amounts of tumour necrosis factor-alpha (TNF-alpha) into the culture medium. Additionally the primary macrophages were coincubated with phosphodiesterase-IV-(PDE-IV)-inhibitors or beta-adrenoceptor agonists that in previous studies demonstrated strong suppressive effects on TNF-alpha release. Especially the beta-adrenoceptor agonists showed to be very potent TNF-alpha suppressants, which indicates that the beta-adrenoceptor might be an interesting target for suppression of activation of macrophages. This was strengthened by the observation that the beta-adrenoceptor expression was not altered during the onset of the inflammatory cascade. PMID- 10682385 TI - Inhibition of aflatoxin M1 production by bovine hepatocytes after intervention with oltipraz. AB - It is well known that cattle ingesting aflatoxin B1 contaminated feed commodities excrete aflatoxin M1 into their milk. As aflatoxin M1 originates from hepatic metabolism, measures to prevent aflatoxin M1 formation need to be directed to either the immobilization of aflatoxin B1 in the gastrointestinal tract or the modification of hepatic metabolism of aflatoxin B1. Here we studied the influence of oltipraz and a second dithiolthione, (1,2) dithiolo (4,3-c)-1,2-dithiole-3,6 dithione (DDD) on bovine hepatic aflatoxin B1 biotransformation. Oltipraz inhibited aflatoxin B1 metabolism as no aflatoxin M1 and no aflatoxin B1 dihydrodiol, the second metabolite found in bovine hepatocytes, was formed. DDD did not significantly inhibit aflatoxin B1 metabolism. It could be demonstrated that the inhibition of aflatoxin B1 metabolism was due to the inhibition of several cytochrome P450 enzyme activities by oltipraz. In contrast, DDD inhibited only ethoxyresorufin O-deethylation activity. These findings suggest a high efficacy of oltipraz in inhibiting aflatoxin M1 contamination of milk from dairy cows exposed to aflatoxin B1 contaminated feeds. PMID- 10682386 TI - Within-farm spread of classical swine fever virus--a blueprint for a stochastic simulation model. AB - A stochastic simulation model to investigate the transmission of classical swine fever (CSF) virus within an infected farm is described. The model is structured according to the processes that occur within and between management groups (pig units or houses). It uses the individual pig as the unit of interest and estimates the number of animals in the states 'susceptible', 'infected', 'infectious', and 'removed' for each day of the disease incident. Probabilities are assigned to the transitions between states. The probability of a pig becoming infected is made dependent on the probability of contact between a susceptible and an infectious pig as well as the probability of transmission. The more pigs become infected in one unit, the more likely is subsequent spread to another management group on the farm. Ultimately, the probability that a shipment of pigs from the farm will include at least one infected pig can be estimated in order to identify high-risk movements during a CSF epidemic. The model results were compared with experimental data on CSF transmission within one pig unit (management group). It could be shown that the model was capable of reproducing the experimentally observed infection and mortality rates. To improve the input parameters and for further model validation, more experimental data and field data from CSF outbreaks are needed. PMID- 10682387 TI - Disease-induced alterations in plasma drug-binding proteins and their influence on drug binding percentages in dogs. AB - Disease-induced variations of plasma albumin (ALB) and alpha 1-acid glycoprotein (AAG) levels were investigated in dogs. Lower ALB (sometimes > 50% reduction) and higher AAG (sometimes > 10-fold increase) levels were observed in dogs with various diseases. Drug binding was determined at therapeutic concentrations using normal, low-ALB and high-AAG dog plasma. The binding percentages of the ALB binding drugs decreased in low-ALB plasma, resulting in a large increase in unbound drug, particularly for naproxen (a 13-fold increase). The binding percentages of all AAG-binding drugs investigated in this study increased in high AAG plasma, resulting in a large decrease in unbound drug, particularly for quinidine (99% decrease). The fluctuation in the unbound fraction of drugs could affect their efficacy or could cause side-effects. Veterinary clinicians should monitor the ALB and AAG levels in the plasma of patients and correct dosage regimens according to these levels, where field conditions permit this, in order to ensure the proper usage of drugs with high affinity for ALB or AAG. PMID- 10682388 TI - Validation of a new method of visual oestrus detection on the farm. AB - Since visual observation is the most commonly used way of detecting oestrus and is supposed to be as effective as detection with technical devices, we evaluated a recently developed oestrus detection scoring system in daily dairy practice. In this scoring system nine signs of oestrus are scored with points, ranging 3 to 100. Twenty-one dairy farmers used the scoring system during a period of 3 weeks. All cows that were more than 30 days post partum and not confirmed pregnant were monitored, using the scoring system, by the herd owners. Oestrus was confirmed by measuring progesterone concentrations in milk. With the scoring system a detection rate of 47% was achieved. This was lower than expected, because in an earlier control period, the detection rate was 64%. We concluded that this new method might be too complicated to introduce to normal herd management, because in daily practice it is too demanding to watch cows twice a day for 30 minutes, especially if the cows show only vague and infrequent symptoms of oestrus. It also appeared to be too complicated to watch the herd at the most appropriate time. However, if the scoring system is included in the daily routine, meaning that farmers are trained to watch for other symptoms than standing heat only and are able to recognize their different values, it can be a valuable aid to oestrus detection. PMID- 10682389 TI - Urinary concentration of corticoids in ponies with hyperlipoproteinaemia or hyperadrenocorticism. AB - The urinary corticoid:creatinine (c:c) ratio was determined in ten pony mares suffering from hyperlipoproteinaemia. The mean (+/- sd) urinary c:c ratio of these ten ponies (47 +/- 31 x 10(-6)) was not significantly different from that of twelve pony mares with a pituitary pars intermedia adenoma (31 +/- 18 x 10( 6). The correlation between the urinary concentration of corticoids and plasma total lipids, and the correlation between the urinary c:c ratio and plasma total lipids in ponies with hyperlipoproteinaemia were not significant (P > 0.05; r = 0.53 and r = 0.008, respectively). Preliminary results favour primary hyperadrenocorticism being associated with hyperlipoproteinaemia. In conclusion, the data presented here suggest that cortisol can contribute to insulin resistance in ponies with hyperlipoproteinaemia. PMID- 10682390 TI - The use of a polypropylene mesh for treatment of ruptured collateral ligaments of the equine metatarsophalangeal joint: a report of two cases. AB - The prognosis of ruptured collateral ligaments of the metacarpophalangeal or metatarsophalangeal joint in horses is usually considered to be poor, especially for future athletic performance. The main problem is the development, due to joint instability, of osteoarthritis, which may result in persistent lameness. In this paper a surgical technique is described in which joint stabilisation is realised by using a polypropylene mesh as a substitute for the ruptured ligaments, with the subsequent application of a cast for 7 weeks. The technique was successfully performed in 2 horses with ruptured lateral collateral ligaments of a metatarsophalangeal joint. Fifteen months after surgery both horses resumed exercise. Performance could be classified as fair in one case and good in the other. It is concluded that the preliminary results obtained with this surgical technique to stabilise ruptured collateral ligaments of the fetlock joint are promising. PMID- 10682391 TI - [Ethics and life: bioethics studies]. AB - A year ago, in June 1998, Diego Gracia published four volumes under the general title of Bioethics and Life: Studies of Bioethics. These volumes are the continuation of two other books published by the same author during the last years: Foundations of Bioethics (1989) and Decision procedures in clinical ethics (1991). The present article is a short presentation of the four volumes written by the author, followed by the transcription of the foreword of the book. Due to the fact that this book is one of the most important contributions made to bioethics in the Spanish speaking world, it will have a very wide influence not only in Spain but also in Latin America. PMID- 10682392 TI - [Determination of allelic losses in peritumor bronchial mucosa]. AB - Lung cancer is the leading of death in both, women and men in the United States and many European countries. molecular cytogenetic and LOH analyses and LOH analyses of non-small cell lung cancer have shown somatic genetic alterations in a variety of chromosomes such as 1p, 3p, 5q, 8p, 9p, 11p, 11q and 17p. Putative tumor suppressor genes have been located in these lci. Allelic loss at 3p21, 9p21 and 5q21 has been also reported in premalignant epithelial lesions of the bronchus. We investigated 33 cases of NSCLC for LOH at 3p, 5q, 9p and 17p: 22 squamous cell and 11 adeno carcinomas. Normal lymphocytes, tumor cells and normal bronquial cells adjacent to the tumor were microdissected from paraffin embedded tissues. PCR amplification was performed utilizing the specific markers D5S299, D5S346, D3S1300, D9S157, D9S171 and D17S799. Our results show that within the NSCLC tumor cells, LOH was more frequently found at the 5q21 locus (75% of the informative cases), the 3p21 locus (48%) and 17p. Within the normal bronquial cells, LOH was found in 33% of the cases at 5q21 and 33% of the cases at 3p21. In conclusion our results show that LOH can be assessed in normal bronquial mucosae adjacent to NSCLC. We suggest that LOH at these loci may be present before the onset of the malignant growth. PMID- 10682393 TI - [Spa therapy and sadness]. AB - The feeling of sadness is said to be an individual problem which can take advantage of an adequate Spa therapy in all its complexity. The sadness as a feeling of disproportional affective reaction with mental consequences, can find relief in the Spa cures and their different structural elements like the hydrothermal techniques, psychical actions, environmental features..., etc. These Spa cures have many favourable effects not only on the body but also on the mind, being capable of liberating the mind of worries and respecting the peculiar personality and way of suffering of the patient. PMID- 10682394 TI - [Arterial grafts]. AB - After a historical review, pointing out the different types of grafts (tubes, veins, arteries, synthetic material), several haemodynamics facts are commented, that in author's opinion, are closely related with biological process of "neo intima" development. Technical aspects are evaluated, including extra-anatomical by-pass, infection risk, and proposed solutions when run-off problems occurred when placing a graft, concluding with modern guidances when using this material, such as endotheliation, cryopreserved artery grafts, polyurethane and pyrolytic carbon grafts, negatively charged grafts, biodegradation prosthesis, laboratory artery development, small diameter grafts and endovascular surgery. PMID- 10682395 TI - [History and perspective of reemerging tuberculosis]. AB - The concepts of infectious emergency, the microbial virulence stimulatory factors, the genetic basis (pathogenicity islands and type III secretion factors) of the microbial variation and adaptation, the interest of animal reservoirs in the infection and transmission cycles, and the reemergence of human and animal tuberculosis are reviewed. In reference to the tuberculosis, the finding of a new species or subspecies of mycobacteria isolated by the authors laboratory from goats and belonging to the important M. tuberculosis group is reported. PMID- 10682396 TI - [Anorectal diseases. Diagnosis and treatment]. AB - The presentation of the book Enfermedades Anorrectales. Diagnostico y tratamiento edited by Dr Fernando Lopez-Rios (Madrid: Harcourt Brace, 1999) consisted of two parts. In the first part, the structure, goals and characteristics of the work were explained by its editor. In the second part, after a brief survey of the history of colorectal surgery, Dr Lopez-Rios discussed the importance of understanding colorectal surgery as a subspecialization of general surgery. PMID- 10682397 TI - [Antepartum brain injuries]. AB - It is generally accepted that congenital brain lesions and especially cerebral paralysis are produced nearly always during the process of delivery. The origin can be hypoxia or obstetrical trauma. This has been true until the 70s, but from that period onwards, the incidence due to the delivery process has been reduced so dramatically, that its appearance can be considered as an accident, as in the umbilical cord prolapsus or during abruptio placentae. More than 80% of the cerebral paralysis occur before delivery due to a variety of causes that are detailed in this report and only the resting 20% can be related to foetal asphyxia, only if the following 4 signs appear: 1 Umbilical artery pH lower than 7.2 Apgar test after 5 minutes of 3 or less. 3 Appearance of a neurological hypoxic-isquemical syndrome. 4 Lesions in another organ that can be attributed to hypoxia. This new international definition indicates, the necessity to put aside the old definition still more generally accepted when a fetus is simply depressed but not with asphyxia. In the same way, the actual concept of fetal suffering during delivery needs also to be rejected not necessarily alterations are followed and can be considered as pejorative. Only if the suffering continues with asphyxia can be accepted. PMID- 10682398 TI - [Epidermolysis bullosa]. AB - The epidermolysis bullosa are a group of genodermatoses in which there is congenital fragility of the skin which produce blisters with the least of traumas. The group includes up to thirty clinical-genetical entities. They are classified according to the level where the blister is found into simplex or epidermolytic (intraepitelial blister), junctional (blister in the dermoepidermal junction) and dystrophic or dermolytic (subepidermal blister) epidermolysis bullosa. The symptoms, classic or Mendelian genetics and the recent findings in the most frequent forms of each one of these groups are reviewed. In most of them, the gene that produces the mutations and the chromosome in which its locus is found are known. In the simplex forms, the disorders lies in the genes that codify the different keratins. In the junctional ones, mutations are found in the laminin genes that act in the development of the anchoring filaments or in those of the other components of the basement membrane. In the dystrophic ones, which are the most serious, the mutations affect the collagen VII gene that codify the development of the anchoring fibrils. There is no treatment for any of the forms of epidermolysis bullosa. It is possible that the future advances in genetic engineering can contribute to their prophylaxis. PMID- 10682399 TI - [Stress and neurodegeneration: pharmacologic strategies]. AB - Long-term exposure to stress has detrimental effects on several brain functions in many species, including humans and leads to neurodegenerative changes. However, the underlying neural mechanisms by which stress causes neurodegeneration are still unknown. We have investigated the role of endogenously released nitric oxide (NO) in this phenomenon and the possible induction of inducible NO synthase (iNOS) isoform. In adult male rats, stress (immobilisation for 6 h during 21 days) increases the activity of a calcium independent NOS and induces the expression of iNOS in cortical neurons as seen by immunohistochemical and Western blot analysis. Three weeks of repeated immobilisation increases immunoreactivity for nitrotyrosine, a nitration product of peroxynitrate. Repeated stress causes NO2(-) + NO3- (NOx) accumulation in cortex, and these changes occurs in parallel with lactate dehydrogenase (LDH) release and impairment of glutamate uptake in synaptosomes. The administration of the preferred iNOS inhibitor aminoguanidine (400 mg/kg i.p. daily from days 7 to 21 of stress) prevents NOx- accumulation in cortex, LDH release and impairment of glutamate uptake in synaptosomes, as well as other markers of oxidative stress such as lipid peroxidation and decrease in glutation. Taken together, these findings indicate that a sustained overproduction of nitric oxide via iNOS expression may be responsible, at least in part, of some of the neurodegenerative changes caused by stress, and support a possible neuro-protective role for specific iNOS inhibitors in this situation. PMID- 10682400 TI - [In memoriam of Dr. Juan Gilbert Queralto]. PMID- 10682401 TI - [They thought; therefore, they spoke]. AB - All functions that characteristically define each form of life are the consequence of the unpredictable process of evolution. The appearance around 300,000 years ago of a complex system of language allowed the homo sapiens to verbally express his thoughts, feelings and experiences in a narrative manner. Thus began the great and continuous development of knowledge and the subsequent post-biological improvement of cognitive functions as a consequence of the stimulation of brain plasticity. This complex process vas not rapid. Important structural and functional craneofacial and pharyngo-laryngeal changes were necessary. Some were already present four million years ago in the australopitecus and became more efficient in homo erectus to reach the definite dimensions of modern humans 125,000 years ago in the homo sapiens-sapiens. From then on a continuous, productive and interactive relationship between the ability to think and the capacity to speak was established. New anonymous ideas invent their corresponding words which, in turn, create more complex sequential thoughts. The fundamental contribution of language in the development of human intelligence raises the question fo which of the two functions preceded the other or whether they both evolved simultaneously. Is it possible to assume that if the pharyngo-laryngeal structures ad not evolved at all, the brain would have invented another type of language? PMID- 10682402 TI - [Our brain deceives us]. AB - The brain does not "objectively" reflect outer reality, but rather what is necessary in order to survive and adapt to the environment. The "phi" phenomenon, the eye's blind spot and cryptomnesia, reflect the brain's capacity to invent information. In split-brain patients, research has shown how one hemisphere, when it lacks certain information, invents a plausible story to explain the behaviour of the other hemisphere. All this would suggest that the brain deceives us, producing occurrences which in reality have never happened, in such a way that, for the brain, convenient reality and "objective" reality are two very different things. PMID- 10682403 TI - Synthesis and analgesic activities of 2-substituted-1H-phenantro [9,10-d] imidazoles. AB - Some 2-substituted-1H-phenantro [9, 10-d] imidazole compounds were synthesized and their analgesic activities were determined on the Swiss albino mice (25-35 g) of either sex. In the synthesis, phenantre-nequinone molecule was reacted with different aldehyde derivatives in acetic acid presence of ammonium acetate and finally 16 compounds were gained. All spectral and elemental analyses of the original compounds were completed and their structures were elucidated. Analgesic activity of the compounds were examined by using the Tail-Clip method. 2-Phenyl 1H-phenantro [9, 10-d] imidazole (comp.1) and 2-(4-chlorophenyl)-1H-phenantro [9, 10-d] imidazole (comp. 6) were found to be more active than the others and their LD50 values were found to be greater than 100 mg/kg (i.p.). PMID- 10682404 TI - Comparative bioavailability of sulfamethoxazole in co-trimoxazole suspensions containing different thickness agents. AB - Many workers have attempted to determine the bioavailability of pharmaceutical formulations, which is important to assure the efficacy and safety of medications. In the present study, we investigated the bioavailability of five formulations of the combination of 0.8% trimethoprim (TMP) and 4% sulfamethoxazole (SMZ) (co-trimoxazole) as a suspension, containing different types of thickness agents. The blood levels of a single oral dose administered to rats were compared. Bioavailability was determined by comparing the time to peak concentration (Tmax), peak serum concentration (Cmax), total area under the concentration time curve (AUC) and the elimination rate constant (Kel). Analysis of the pharmacokinetic parameters of SMZ showed significant differences between the formulations, indicating that the absorption of SMZ was affected by thickness type. The calculated bioavailabilities of oral TMP and SMZ were 381, 558, 695, 480, 559 and 554 micrograms/mL, respectively, and the preparation containing hydroxyethyl cellulose 4.400 H as a thickness agent showed the best bioavailability (AUC 0-infinity = 695.24; micrograms/mL; Cmax = 35.2 micrograms/mL). PMID- 10682405 TI - Albumin microsphere as a drug delivery system for dexamethasone: pharmacokinetics in sheep, residue amount in cows and distribution in rats. AB - The aim of this study was to evaluate the potential use of albumin microspheres as a drug delivery system to provide sustained release of dexamethasone in vivo. Pharmacokinetic studies were carried out in sheep and tissue distribution in rats given dexamethasone in two injectable forms--water suspension of dexamethasone associated with albumin microspheres and water-ethanol solution of dexamethasone. When dexamethasone was associated with albumin microspheres the amount of free dexamethasone, available for absorption did not reach the high values typical of the pattern of release of dexamethasone from water-ethanol solution. The time of withdrawal in lactating cows after i.m. administration of DXM-AM was found to be 5 days. The release of dexamethasone from the albumin microspheres was accomplished gradually thus allowing sustained levels of the corticosteroid to be maintained for several days in blood plasma, milk, liver, kidney and muscles. PMID- 10682406 TI - Physico-chemical activity relationship of some substituted benzimidazoles. PMID- 10682407 TI - Assessment of acute cyto- and genotoxicity of corrosion eluates obtained from orthodontic materials using monolayer cultures of immortalized human gingival keratinocytes. AB - Whilst a patient is undergoing orthodontic treatment, dental appliances based on non-precious metals or titanium remain in the oral cavity for up to several years. Throughout this period the appliance is in either direct or indirect contact with the oral mucosa. To investigate the possibility of cell damage occurring as a result of appliance corrosion, monolayer cultures of immortalized human gingival keratinocytes were assessed for acute cyto- and genotoxicity using the hexosaminidase assay and the Comet assay respectively. The materials tested included 1. a nickel-free wire, 2. a UK-1 bond, 3. nickel-free as well as nickel containing brackets with and without color signature and 4. a titanium expansion screw. Each of the test materials was corroded in a solution consisting of equal amounts of lactic acid and sodium chloride (0.1 M) for 1, 3, 7 and 14 days. The cell cultures were then exposed to eluates exhibiting the highest ion concentrations. None of the eluates was found to exhibit acute cytotoxicity, regardless of the type of test system used. Qualitative assessment using neutral red dye for live cells and either trypan blue or propidium iodide to disclose dead cells failed to reveal any significant increase in cell damage when exposed cells were compared to control cultures. Unrestricted cell vitality was confirmed by quantifying viable cells through measurement of hexosaminidase enzyme activity. Furthermore, assessment of genotoxicity revealed no apparent DNA damage to immortalized gingival keratinocytes following exposure to the test eluates. Because the materials tested in this study were corroded using the exacting methods normally applied to precious metals or gold-containing alloys, the lack of either acute cyto- or genotoxic effects following exposure to the test eluates indicates that the materials tested exert no adverse effects on cells similar to those of the target tissue exposed to the materials in situ. PMID- 10682408 TI - A longitudinal study on growth in untreated children with Angle Class II, Division 1 malocclusion. AB - The aim of this longitudinal study was to analyze growth-related changes in untreated Class II, Division 1 subjects. Forty untreated Swedish children with Angle Class II, Division 1 malocclusion were cephalometrically examined. The average age of the total group was 10.1 years at the first examination and 12.0 at the second. In addition to statistical comparisons of the average values of variables at both time points, multivariate analyses (harmony box according to Hasund and Segner, regression equation analyses) were also performed. This permitted individual evaluation of skeletal structures, position of incisors, and the soft tissue profile relative to the given craniofacial configuration. With the ANB angle remaining unchanged, both jaws were transposed ventrally. The reduction of the angle between the palatal and mandibular planes and the gonial angle was accompanied by anterior rotation of the mandible. The lower incisors protruded, and upper and lower first molars exhibited a translation to ventrocaudal. According to harmony box analysis, the most frequent cause of a distal relationship between the 2 jaws was a disharmoniously anterior shift of the maxilla. This was confirmed by the additional analysis of the available unpublished raw data from untreated Norwegian and Munich Class II/1 subjects and from other studies. In such cases, orthodontic treatment methods exerting a growth-inhibitory effect on the maxilla should be used. PMID- 10682409 TI - Facial profile and dental changes before, during and after treatment with Hansaplate/Headgear. AB - In this prospective study the changes of facial profile and dentition in 19 boys and 19 girls treated for 1 year with Hansaplate/Headgear were analyzed yearly over a 4-year period. On average the boys were 10.2 and the girls 9.2 years old at the first recording, i.e., 1 year pretreatment. The actual values are compared with standard growth data presented by Bathia and Leighton (1993). Irrespective of the initial face morphology, the Hansaplate/Headgear appliance straightened the facial hard and soft tissue profiles favorably. The upper lip became more retruded while the lower lip was unaffected by therapy. During the 2-year post treatment period these changes continued, probably as a consequence of growth. The overjet was reduced by 5 mm during treatment and was found to be quite stable 2 years after treatment. PMID- 10682410 TI - Axiographic evaluation of mandibular mobility in children with angle Class-II/2 malocclusion (deep overbite). AB - Correction of the occlusion in Angle Class-II/2 patients is often more complicated and tedious than in Class-II/1 cases. Reasons given are the more comprehensive remodeling and functional adaptation processes of the temporomandibular joints (TMJ) since the articular tuberculum is more strongly developed in deep overbite, and a deep bite leading to a locked occlusion is dominant due to the steeper condylar path. Both characteristics are, however, considered to be the consequence of pronounced incisor retrusion. The present study covered 28 untreated Class-II/2 patients aged 8 to 12 years in whom functional TMJ adaptation to the retroclined maxillary incisors was studied with the help of electronic, 3D axiographic registrations of mandibular movements. Comparison with eugnathic age peers revealed increased mobility in mandibular protrusion and a somewhat steeper condylar path, although the latter was less pronounced than in adult patients. The results corroborate the concept of functional TMJ adaptation to incisor inclination and speak for early uprighting of maxillary incisors. PMID- 10682411 TI - The influence of a 0.2% chlorhexidine mouthrinse on plaque regrowth in orthodontic patients. A randomized prospective study. Part I: clinical parameters. AB - In a prospective plaque regrowth study focusing on oral hygiene during fixed appliance therapy 12 adolescent patients (mean age 14.1 +/- 1.5 years) were evaluated twice over 2-day test periods. In the randomized, double-blind study the influence of a 0.2% chlorhexidine (CHX) mouthrinse (Corsodyl) and a commercially available dentifrice supplementing fluoride (Odol-med-3) were compared intra- and interindividually in a crossover design with regard to plaque and gingivitis. Before starting the first test phase there was a 14-day preliminary phase for upgrading the oral hygiene. Between the 2 test phases was a 5-day "washout". On the last day of the second test phase the patients were asked to fill in a questionnaire concerning their experiences during the study. The 0.2% Corsodyl reduced the plaque index scores significantly (p < 0.001). The gingival index revealed a similar reduction (2nd day of test: p = 0.03). Until the 5th day of washout a clear-cut carryover effect of the chlorhexidine rinse on the gingival index was observed. Both the lower mean values of the 2 clinical parameters at the beginning of the test phases as compared with those at the beginning of the preliminary phase and the evaluation of the questionnaires indicated a possible Hawthorne effect. 0.2% Corsodyl may be employed as an adjunct to other preventive measures such as professional care and patient oriented instruction on an intermittent basis in order to reduce the plaque induced iatrogenic side effects and to enhance the efficacy of oral hygiene measures in connection with orthodontic therapy with fixed appliances. PMID- 10682413 TI - MedBytes. PMID- 10682412 TI - More tort reform is needed. PMID- 10682414 TI - Insuring the children. PMID- 10682415 TI - Measuring the mandates. PMID- 10682416 TI - Living longer & better! PMID- 10682417 TI - Traditions and transformations. PMID- 10682418 TI - Telemedicine in Texas. PMID- 10682419 TI - Area income as a predictor of preventable hospitalizations in the Harris County Hospital District, Houston. AB - This study assesses whether geographical area income based on census data is a good predictor of preventable (or ambulatory care-sensitive) hospitalizations in a large public hospital system in Texas, and how area income correlates with the socioeconomic status reported by patients. Documenting a correspondence between area and individual-level income, as well as meaningful variation in rates of avoidable hospitalizations across subgroups and areas with varying concentrations of low-income persons, points to the validity and utility of this approach for monitoring how well public hospital systems in Texas address the needs of those they most directly serve. Area income may not fully reflect the disproportionately lower socioeconomic status of patients seen in the public hospital system. Nonetheless, living in lower-income zip codes was associated with higher preventable hospitalization rates for the predominantly low-income population served by the public hospital system. A tenfold difference found in the adjusted rates of hospitalizations for preventable conditions compared with control (or marker) conditions among persons living in low-income areas signals the likelihood of substantial unmet needs in this population. Small-area analysis and related comparisons of rates of preventable hospitalizations in high- and low income areas provide useful indicators for monitoring and assessing the performance of public hospital systems in Texas. PMID- 10682420 TI - [Study on the mechanism for fetal asphyxia in intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the cause of fetal asphyxia of mothers with intrahepatic cholestasis of pregnancy (ICP). METHODS: The cord plasma erythropoietin (EPO) concentrations were measured in 37 infants of mothers with ICP and 46 control infants after elective cesarean section. Furthermore, the transfer of oxygen across the placental membranes in ICP group (n = 7) were compared with controls (n = 8) by dual perfusion of the human placental lobule in vitro. RESULTS: The oxygen transfer across the placental membrane in ICP was similar to the controls (P < 0.05). However cord venous EPO was lower in infants delivered by elective cesarean section from women with ICP (13.58 +/- 8.88 IU/L, P < 0.05) than that in control infants (20.43 +/- 14.15 IU/L). CONCLUSION: The transfer of oxygen across the placental membrane in ICP may be normal. The lower cord venous EPO value in ICP may be mainly responsible for the fetal asphyxia. PMID- 10682421 TI - [Influence of prepregnancy weight and maternal weight gain on pregnancy outcome]. AB - OBJECTIVE: To determine the influence of weight before pregnancy and maternal weight gain on pregnancy outcome. METHODS: Body wight index (BWI) was calculated before pregnancy and maternal weight gain and pregnancy outcomes were followed up in 2,584 primipara with singleton births. RESULTS: Compared with normal weight women, the prevalences of pregnancy induced hypertension, operative deliveries and high birth weight infants were significantly higher in women with heavy weight, and prevalence of low birth weight infants was significantly higher in women with lower weight. CONCLUSION: Prepregnancy weight and weight gain during pregnancy have direct impact on birth weight of the baby and maternal complications. PMID- 10682422 TI - [Detection and enrichment of fetal cells in maternal circulation for prenatal diagnosis of fetal sex]. AB - OBJECTIVE: To study fetal cells from maternal peripheral blood for prenatal diagnosis of fetal sex. METHODS: Samples of peripheral blood in 64 women of 8-40 gestational weeks were collected to enrich the fetal nucleated red blood cells by density gradient centrifugation. DNA was extracted from each samples of enriched fetal cell for PCR amplification of Y chromosome specific DNA to determine fetal sex. RESULTS: Fetal nucleated red cells were found in 25 out of 64 maternal samples (39.06%). Y chromosome 149 bp was found in 28 cases of the 33 mothers given birth to male babies. One Y specific DNA sequence was detected in 31 women had female babies. The sensitivity was 84.85% and specificity 96.77%. The overall agreement of the diagnosis was 90.63%. CONCLUSION: Density gradient centrifugation can enrich fetal nucleated red blood cells from maternal peripheral blood. Fetal sex can be determined by PCR amplification of Y chromosome specific DNA with these fetal cells. PMID- 10682423 TI - [Immuno-pathomorphological study of placenta with anticardiophospholipid antibody positive]. AB - OBJECTIVE: To investigate the relationship between the anticardiophospholipid antibody (ACA)-positive cases and the poor pregnancy outcomes by observing the immuno-pathomorphology of the placenta. METHODS: 16 cases of ACA-positive placenta (8 normal and 8 poor pregnancy outcomes) were collected as the studied group, and 8 cases of ACA-negative placentas served as the controlled group, and immunofluorescence technique (IFT) was used. RESULTS: In the studied group, the deposits of immuno-complex were found in the cytoplasm of trophoblast and on the walls of villous blood vessels. IgA manifested as filiform, and IgG, IgM, C3 and C4 appeared as filiform, granular and tubercular as well. In contrast, all the immuno-globulins displayed negative reaction in the controlled group, and C4 demonstrated some small focal positive reaction. The frequency of ACA-positive in IUGR cases were 19.30% (22/144), and of the IUGR in all the ACA-positive pregnant women was 15.28%(22/144). There were 3 cases of IUGR complicated by severe pregnancy induced hypertension, 2 cases of prematurity and 5 newborns with serum ACA IgG positive. CONCLUSION: The poor pregnancy outcome in ACA-positive women is related to the dysfunction of autoimmuno-system, and ACA-positive may be one of the causes of IUGR. The immuno-injury occurred in the placenta may affect the normal function of placenta. Maternal antibody may be transmitted to the fetus, and its significance needs further study. PMID- 10682424 TI - [Changes of carbohydrate metabolism in normal pregnancy and its relationship with placental lactogen concentrations]. AB - OBJECTIVE: To observe the changes of glucose metabolism and its relationship with human placental lactogen (HPL) in normal pregnant women. METHOD: 94 normal pregnant women had their serum HPL concentration tested and had undergone a 75 g oral glucose tolerance tests (OGTT). Both serum insulin and glucose levels at different time intervals were determined, and the areas under curve (IAUC, GAUC) were caculated, also insulin resistance (IAI) was evaluated. RESULTS: GAUC, IAUC, IAI and HPL are all significantly increased with advance of gestation (P < 0.05). Multiple correlation coefficient study demonstrated that HPL is significantly positively correlated with GAUC, IAUC and IAI(P < 0.001). But when IAUC and IAI were controlled, the relationship between HPL and GAUC was not found. CONCLUSION: There are hyperinsulinemia and insulin resistance during pregnancy, and they are related to HPL. HPL is one of the factors that contributing to carbohydrate metabolism changes during pregnancy. PMID- 10682425 TI - [Study on a deletion polymorphism of the angiotensin converting enzyme gene in pregnancy induced hypertension]. AB - OBJECTIVE: To study a polymorphism and allele frequency of the angiotensin converting enzyme (ACE) gene in pregnancy induced hypertension (PIH). METHODS: Polymerase chain reaction (PCR) for detection of ACE gene polymorphism was performed to show the deletion/insertion (D/I) polymorphism in intron 16 of ACE gene. A 490 bp(I) and 190 bp(D) PCR products were identified, corresponding to the PCR amplification of the the allele with or without the insertion. RESULTS: Derived allele frequencies for insertion and deletion were the different between 35 PIH and 25 control subjects. Compared the frequency of D allele gene (0.76) and the percentage of the ACE DD genotype (65.7%) in the PIH patients with the frequency (0.28) and the percentage (8.0%) in the control population, they were significant higher in individuals with PIH (P < 0.001). CONCLUSIONS: There was an excess of DD genotype and the frequency of D allele gene in PIH, confirming the genetic variation in the ACE locus could be involved in the risk of PIH and suggesting the ACE gene may contribute to the pathogenesis of PIH. PMID- 10682426 TI - [A study on the relationship between serum anticardiolipin antibody and embryo implantation in an in vitro fertilization and embryo transfer program]. AB - OBJECTIVE: To study the role of serum anticardiolipin antibody in embryo implantation in an in-vitro fertilization and embryo transfer program. METHODS: 150 in vitro fertilization and embryo transfer (IVF-ET) treatment cycles from Nov. 1994 to May, 1996 were studied. Serum anticardiolipin antibody was measured by enzyme-linked immunosorbent assay (ELISA), and the clinical pregnancy outcome observed. RESULTS: Lower pregnancy rate 9.5% was found in 21 cycles of anticardiolipin antibody seropositive women compared with 26.3% in 129 cycles of anticardiolipin antibody seronegative women (P < 0.05). Patients with biochemical pregnancy and no pregnancy had seropositive anticardiolipin antibody in 20.0% and 16.2% compared with 5.6% in the patients with clinical pregnancy (P < 0.05). CONCLUSIONS: Failure of embryo implantation is associated with many factors. Our study demonstrated the serum anticardiolipin antibody in patients may play a part in embryo implantation and very early postimplantation loss. PMID- 10682427 TI - [Pathological and immunohistochemical study on estrogen and progesterone receptors in endometrium of polycystic ovarian syndrome]. AB - OBJECTIVE: To investigate the changes of endometrium in infertility patients with polycystic ovarian syndrome (PCOS). METHODS: The endometrium from 39 patients with PCOS were studied by HE stain and immunohistochemical method using anti estrogen receptor(ER) and anti-progesterone receptor(PR) monoclonal antibodies. RESULTS: 87.2% (34/39) of the endometrium showed anovulatory proliferative phases; 51.3% (20/39) had signs of hyperplasia; 35.9% (14/39) showed asynchronization of endometrial glands and 46.2% (18/39) poor response of stroma to sex hormone. The levels of ER and PR in endometrium during proliferative phase were significantly higher than those of normal control, and the PR in the stroma of hyperplasia endometrium had an uneven distribution and was significantly less than that in the control group (P < 0.05). CONCLUSIONS: The endometrial histopathological changes and local decrease or absence of ER and PR in PCOS might be one of the causal factors of infertility. PMID- 10682428 TI - [The clinical value of tissue polypeptide antigen in ovarian carcinoma]. AB - OBJECTIVE: To evaluate the clinical usefulness of tissue polypeptide antigen (TPA) in diagnosis and monitoring the course of patients with ovarian carcinoma. METHODS: Serum levels of TPA and cancer antigen 125 (CA125)were measured by radioimmunoassay in 60 patients with advanced ovarian carcinoma (41 with active disease and 16 with nonactive, 3 with metastatic tumor from gastrointestinal carcinoma), 24 healthy women and 27 benign gynecologic tumors. RESULTS: The serum TPA and CA125 levels were elevated in 82% and 70% of patients with ovarian carcinoma respectively, the total positive rate of the two markers was 92%. TPA and CA125 levels were within normal range among most of healthy women and patients with benign tumors. TPA and CA125 levels were also correlated with the regression or progression of the disease. CONCLUSION: Combined measurement of TPA with CA125 was useful for the differential diagnosis and achieving higher positivity in patients with epithelial ovarian carcinoma. PMID- 10682429 TI - [The expression of glutathione S-transferase pi in human ovarian cancer as an indicator of resistance to chemotherapy]. AB - OBJECTIVE: To study the relationship between the expression of glutathione S transferase pi (GST-pi) in cancer tissue and the chemoresistance in patients with ovarian carcinoma. METHODS: The expression of GST-pi in 53 cases of ovarian cancer, 20 ovarian benign tumors and 17 normal controls was determined by using immunohistochemical SP method and the results were studied in correlation with some clinical and pathological data. RESULTS: (1) Positive expression with GST-pi was demonstrated in 60.4% of malignant and 10.0% of benign ovarian tumors while the expression was negative in normal controls. (2) No significant difference for the expressions was shown in relation to the histopathology, the clinical staging and the size of residual tumors after cytoreductive surgery. (3) The positive rate of the expression was 47.2% (17/36) for the initially treated patients while those with tumor recurrence had a positive rate of 88.2% (15/17). (4) GST-pi positive cases showed less response rate of 37.5% (12/32) to chemotherapy as compared with that of 76.2%(16/21) for GST-pi negative cases. (5) The survival period of the patients with GST-pi positive expression was shorter than that of those with GST-pi negative expression. CONCLUSION: The expression of GST-pi in patients with ovarian carcinoma in closely related to the chemosensitivities clinically. Determinations of the GST-pi are useful for predicting the chemosensitivities and the prognosis of the disease. PMID- 10682430 TI - [Relation between HLA antigen system and pathological pregnancies]. PMID- 10682431 TI - [Prevention and treatment of pregnancy-complicated tuberculosis]. PMID- 10682432 TI - [Ovarian cancer treated with very-high-dose chemotherapy and peripheral hematopoietic stem cell transplantation]. PMID- 10682433 TI - [Detection of human papillomavirus infection in cervical Pap smears by computer assisted cytologic test]. AB - OBJECTIVE: The sensitivity and accuracy of cytologic computer-assisted test (CCT) in diagnosis of human papillomavirus (HPV) infection in cervical Pap smear were evaluated. METHODS: Cervical Pap smears obtained from 158 patients with vulva condyloma acuminata were examined by CCT. The diagnostic criteria were based on The Bethesda System (TBS). Simultaneously HPV DNA in cervical mucus was examined by polymerase chain reaction (PCR) technique and cervical biopsy taken under the guidance of colposcopy for pathological examination. RESULTS: The sensitivity and accuracy of CCT in diagnosis of cervical HPV infection were 74.62% and 67.72% compared with pathological results whereas 69.11% and 70.88% compared with PCR results respectively. CONCLUSION: The results indicates that CCT, a cytologic pathological technique, is a useful method in diagnosis of clinical and subclinical cervical HPV infection. PMID- 10682434 TI - [The evaluation of computer cytological test with colposcopy for the diagnosis of cervical lesions]. AB - OBJECTIVE: To assess the value of computer cytological test (CCT) with colposcopy for the diagnosis of early cervical diseases. METHODS: 3,016 cervical pap smears have been done in our hospital from March 15, 1995 to March 15, 1996. The CCT smears were analyzed first, then cytologist made the final reports, cervical biopsies under the guidance of colposcopy were taken in 56 cases with abnormal findings. RESULT: We found 375 (12.4%) abnormal pap smears out of 3,016 cases, in which cervical squamous cancer 5 cases, endometrium adenocarcinoma 1 case. The low grade squamous intraepithelial lesion (LSIL) and human papilloma virus (HPV) infection most frequently happened at the younger patients. There are 59 cases (2.0%) of LSIL, among which HPV infection accounted 79.7%. We use CCT for primary selection, its sensitivity is 98.9% specificity is 90.9%. CONCLUSION: Sexually transmitted diseases is closely related to cervical early disease. CCT should be done in the younger women, especially high-risk patients. Positive cases of CCT must be examined by colposcopy and pathological biopsy, this is the best way to diagnosis cervical disease. PMID- 10682435 TI - [Effects of mifepristone on proliferation and apoptosis in early chorionic villi]. AB - OBJECTIVE: To study the effects of mifepristone on proliferation and apoptosis in early pregnant chorionic villi. METHODS: Proliferation and apoptosis in early pregnant choionic villi from surgical aspiration and mifepristone induced abortion were studied. Marker for proliferation is proliferating cell nuclear antigen (PCNA) immunohistochemical staining, apoptotic cell death was detected using DNA in situ terminal deoxynucleotidyl transferas-mediated dUTP-biotin nick ending labeling (TUNEL). RESULTS: In early pregnant chorionic villi, proliferating index of cytotrophoblasts was 53.74% +/- 1.34%, not only nuclear but also cytoplasma were PCNA positive staining, syncytiotrophoblasts were negative. There existed apoptotic cell death in trophoblasts, the apoptotic index were 2.52% +/- 0.86% in syncytiotrophoblasts and 0.52% +/- 0.26% in cytotrophblasts, respectively. After 2 days mifepristone treatment, the proliferating index of cytotrophoblasts decreased slightly and PCNA positive cytoplasmic expression disappeared; while apoptotic cell increased significantly, the apoptotic indices were 22.16% +/- 2.26% in syncytiotrophoblasts and 20.12% +/ 1.74% in cytotrophoblasts, respectively. CONCLUSION: Mifepristone may inhibit proliferation and promote apoptosis of trophoblasts in early pregnant chorionic villi. PMID- 10682436 TI - [Effect of mifepristone on the concentration of epidermal growth factor in serum and villi of early pregnant women]. AB - OBJECTIVE: To investigate the change of epidermal growth factor (EGF) concentration in serum and villi of early pregnant women and its possible effects by mifepristone. METHODS: Twenty normal women as control and sixty-six early pregnant women were enrolled for study. Serum EGF, estradiol (E2) and progesterone (P) concentrations were measured with radioimmunoassay. EGF concentration in villi by surgical aspiration and mifepristone induced abortion were compared. RESULTS: Serum EGF concentration in early pregnant women was significantly higher than that in non-pregnant women (P < 0.01). The serum EGF, E2 and P concentrations increased as pregnancy advanced (P < 0.05). The growth rate of gestational sac declined and serum EGF and P concentrations decreased slightly after administration of mifepristone. Villi EGF content in mifepristone group was significantly lower than that in surgical group (P < 0.05). CONCLUSION: EGF may be involved in regulating embryo development. Mifepristone may interfere embryonic development via inhibiting EGF level. PMID- 10682437 TI - [A study on the relationship between hemorheology of pregnant women with pregnancy induced hypertension and hemodynamics of fetal umbilical artery blood flow]. PMID- 10682438 TI - [Ultrasonic measurements of fetal thigh soft tissue thickness in the estimation of fetal weight]. AB - OBJECTIVE: To study ultrasonic measurement of fetal thigh soft tissue thickness in the estimation of fetal weight. METHODS: Fetal biparietal diameter (BPD), head circumference (HC), abdominal circumference (AC), femur length (FL) and fetal thigh soft tissue thickness (FTSTT) were measured by ultrasonography and analyzed with neonatal birth weight in 178 cases. RESULTS: There was significant correlation between FTSTT and neonatal birth weight (r = 0.8601), and it is better to estimate fetal weight with FTSTT than with the other parameters. The sensitivity and specificity for detection of macrosomia was 91% and 94%, respectively. The FTSTT was positively correlated with gestational age (r = 0.7070). CONCLUSION: The ultrasonic measurement of FTSTT is a simple, accurate and valuable index in estimation of fetal weight. PMID- 10682439 TI - [Effects of human chorionic gonadotropine and insulin on androgen production by cultured thecal cells from patients with polycystic ovary syndrome]. PMID- 10682440 TI - [Application of multi-tumor markers in ovarian carcinoma]. AB - OBJECTIVE: To improve the specificity and sensitivity of diagnosis and to strengthen postoperative monitoring for patients with ovarian cancer. METHODS: Sera obtained from patients-ovarian epithelial carcinoma (67 cases), benign ovarian tumor (33 cases) and from donor (38 cases) as control. Serologic examination of 5 tumor markers--SA, LSA, CA125, CP2, and 6B11Ab2 was performed. RESULTS: The sensitivity and specificity in diagnosis of ovarian cancer were 83.6% and 85.9% respectively for CA125 alone (> 35 kU/L) whereas 86.6% and 94.4% respectively for multi-tumor markers combined in which 3 or more indices showed positive. In addition, serial measurements of multi-tumor markers have been done for 1 year after operation in 24 cases of ovarian epithelial carcinoma. The correlations between the levels of multi-tumor markers either and the results of second look operation or and clinical manifestation of recurrence were analyzed. CONCLUSION: Multi-tumor markers examination could improve the diagnosis of ovarian cancer and early detection of recurrence. PMID- 10682441 TI - [Study on multidrug resistant gene (MDR1) expression between neoplastic cells and peripheral blood lymphocytes in ovarian carcinoma]. AB - OBJECTIVE: To explore the correlation of MDR1 (P-glycoprotein, P170) gene expression P170 contents among neoplastic tissues, peripheral blood lymphocytes (PBL) and ascites cancerous cells (ACC) during chemotherapy. METHODS: Content of P170 in cancer tissue, ACC and PBL was quantiated dynamicly by using flow cytometric-immunologic method in 48 cases suffered from ovarian carcinoma. RESULTS: Cancer tissue showed positive P170 in 25% of patients. During chemotherapy, P170 content in PBL and ACC were statistically increased. A significant correlation of P170 was found between PBL and ACC (0.01 < P < 0.05). CONCLUSION: The results indicated that multidrug resistance was produced gradually during chemotherapy. Content of P170 in PBL could probably be used as an indirect index of multidrug resistance for recurrent carcinoma or residual tumor. PMID- 10682442 TI - [A clinicopathological analysis of 15 cases with congenital tumors in fetus and newborn]. AB - OBJECTIVE: The incidence rate of congenital tumors in fetuses and neonates and its influence on fetal outcome including development and death were analysed. METHODS: Both clinical manifestations and pathological data of 15 cases with congenital tumor were analysed retrospectively. RESULTS: The incidence rate of congenital tumors in fetuses and neonates was 7.7/100,000 with a rate of 0.7% in total perinatal autopsies. The most common tumor was teratoma (46.7%), the next was hemangioma (26.7%). Among 15 cases with congenital tumor, 9 cases (60.0%) were complicated with polyhydroamnios; 11 perinates (73.3%) were born with body weight appropriate for gestational age, 6 perinates (40.0%) were associated with various anomalies which were commonly secondary to the tumors. The common causes responsible for death were malformation, placental impairment due to tumor invasion or tumor grown at a special site. CONCLUSION: The incidence rate of congenital tumors in fetuses and neonates was very low. Because the tumor usually exhibited a local effect, under carefully evaluating the development of neonates, it is promising to get complete cure. PMID- 10682443 TI - [Advances in the mass screening of Down's syndrome in early and middle stages of pregnancy]. PMID- 10682444 TI - [Risk factors for fallopian tube pregnancy]. PMID- 10682445 TI - [Inhibin and ovarian neoplasms]. PMID- 10682446 TI - [Advance in reproductive immunology]. PMID- 10682447 TI - [The role of decidua tissue lymphocytes in early pregnancy]. AB - OBJECTIVE: To investigate the role of lymphocytes in early pregnancy decidua tissue. METHODS: Immunohistochemical and in situ hybridization techniques were employed to demonstrate the population, phenotype and mediator of lymphocytes in early pregnancy decidua. RESULTS: In early pregnancy decidua, CD8+ suppressor/cytotoxic T lymphocytes were very few. CD4+ helper lymphocytes were few and CD56+ lymphocytes (also named NK-like cell) were abundant. CD56+ lymphocytes containing perforin protein and expressing perforin mRNA were observed. CONCLUSIONS: The lymphocytes in early pregnancy decidua are specific and may play an important role in early pregnancy. They probably exert some positive influence on embryo implantation and placental development, and maintain the balance between trophoblastic cells and decidua tissue. PMID- 10682448 TI - [Relationship between pregnancy loss and antiphospholipid antibody]. AB - OBJECTIVE: To investigate the relationship between history of pregnancy loss and antiphospholipid antibodies (APA), including anticardiolipin (ACA) and lupus anti coagulant antibodies (LA). METHODS: Levels of serum APA, ACA and LA were determined by enzyme linked immunoabsorbent assay and activated partial thromboplastin time method respectively in 122 patients with history of unexplained pregnancy loss (study group) and 100 normal nonpregnant women (control group). The study group was further divided into three subgroups: embryo growth arrest (n = 23), stillbirth (n = 31), and recurrent abortion (n = 63). RESULTS: The positive rates of APA, ACA, LA in the study group were significantly higher than those in controls (P < 0.05-0.001). The above significance was true in both stillbirth and recurrent abortion groups but not in embryo growth arrest group. CONCLUSIONS: Levels of serum APA were associated with pregnancy loss, especially recurrent abortion and stillbirth. We suggested routine screening of serum APA should be performed in patients with history of fetal wastage for the sake of early treatment. PMID- 10682449 TI - [The study of autoantibodies in women with habitual abortion of unknown etiology]. AB - OBJECTIVE: To evaluate the relationship between habitual abortion of unknown etiology and autoantibodies. METHODS: IgG and IgA antibodies against cardiolipin, specific antinuclear double-stranded DNA and single-stranded DNA were measured by enzyme linked immunosorbent assay (ELISA). Antinuclear antibody, smooth muscle antibody, antimitochondrial antibody and antiheart muscle antibody were performed by indirect immunofluorescence assay. Antiextractable nuclear antigen's antibodies were determined by immunodiffusion assay. RESULTS: Most women with habitual abortion of unknown etiology showed higher incidence of antibodies to cardiolipin, nuclear, smooth muscle compared with normal multigravida control (P < 0.05). CONCLUSIONS: These results indicate that most women with habitual abortion of unknown etiology have a higher level of autoreactivity, which may affect the course of pregnancy. The incidence of antismooth muscle antibody in women with habitual abortion of unknown etiology is also increased compared with normal multigravida control. PMID- 10682450 TI - [Alteration of peritoneal lymphocyte transformation and its interleukin-2 release in patients with infertility and endometriosis]. AB - OBJECTIVE: To study the relationship between alteration of peritoneal lymphocyte transformation activity, its interleukin-2 (IL-2) release and infertility associated with endometriosis. METHODS: Peritoneal fluid was collected from 15 infertile patients with endometriosis (endometriosis group) and 13 normal fertile women (control group). Lymphocyte transformation activity were determined in peritoneal fluid by morphometry. IL-2 contents were determined in peritoneal fluid and supernatants of cultured pelvic lymphocytes by biological method. RESULTS: Lymphocyte transformation activity reinforced (P < 0.001) and IL-2 contents increased in peritoneal fluid and supernatant of cultured pelvic lymphocyte in the endometriosis group than those in the control group (P < 0.01, P < 0.001 respectively). There is significant positive correlation between lymphocyte transformation rate and IL-2 activity of cultured supernatant in endometriosis group (P < 0.001). CONCLUSION: The increase of IL-2 content in the samples of endometriosis may play an important role in infertility with endometriosis. PMID- 10682451 TI - [Experimental study of effects of anti-ovarian antibodies on ovarian histology and function]. AB - OBJECTIVE: To investigate the effects of anti-ovarian antibodies (AOA) on ovarian histology and functions. METHODS: Rabbit antibodies against mice ovarian tissues were obtained by immunizing with mice ovarian extracts and purified. The effects of AOA on mice ovarian histology were examined under light and electronic microscope. In addition, changes of ovarian functions, including natural pregnant rate and pregnant mare serum gonadotropin (PMSG) + hCG induced ovulation rate and pregnant rate, were also observed. RESULTS: After treating with different doses of AOA pathological changes occurred in a variety of ovarian components, especially in zona pellucida and granulosa cells, which is more serious in high AOA level group than that in low AOA level group. The natural pregnant rate decreased to zero in both AOA groups. The ovulation and pregnant rate induced by PMSG-hCG were significant lower in both AOA groups than that in the control group especially in the high AOA level group. CONCLUSION: AOA may damage the ovarian tissues and reduce the ovulation and pregnant rate. The higher the AOA level, the more serious pathological changes and the poorer curative effects of PMSG-hCG may occur. PMID- 10682452 TI - [The study on relationship between fetal blood flow velocity waveforms and cord blood gas analyses in normal full term pregnancy]. AB - OBJECTIVE: To study the relationship between fetal blood flow velocity waveforms and cord blood gas values in normal full term pregnancy. METHODS: Fetal umbilical artery (UA) and middle cerebral artery (MCA) and abdominal aorta (AbAo) flow velocity waveforms were measured in 45 full term fetuses 24 hrs before selective cesarean section. Resistance index (RI) and pulse index (PI) were calculated, cord blood samples were obtained from the umbilical artery at cesarean section and blood gas values were measured immediately. RESULTS: (1) UA RI had a significant negative correlation with pH and PO2, and a positive correlation with PCO2. (2) MCA RI had a significant positive correlation with pH and PO2, and a negative correlation with PCO2. CONCLUSION: A Doppler study of fetal blood flow velocity waveforms would be a useful method in the evaluation of the fetal status in the uterus. PMID- 10682453 TI - [Lipoprotein lipids in polycystic ovarian syndrome: independent associations with androgen excess and insulin resistance]. AB - OBJECTIVE: To investigate the relationship between androgen excess, insulin resistance and altered lipoprotein lipids in polycystic ovarian syndrome (PCOS). METHODS: Both basal and luternizing hormone-releasing hormone (LHRH, 100 micrograms) induced responses of circulating testosterone (T), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C) and apoprotein A1 (apo A1) were measured in two PCOS groups with similar T levels (LH/FSH > or = 3, Group 1, n = 15, LH/FSH < 3, Group 2, n = 15) and the control (n = 20) of matched body mass index with Group 1. Insulin resistance was assessed by fasting insulin levels. RESULTS: In basal state, the mean TG levels in women of three groups were as follows: Group 2 > Group 1 > control, while HDL-C showed the opposite. After LHRH test, mean T and TG concentrations increased and HDL-C decreased in all PCOS subjects, especially Group 2 whereas the indices remained unchanged in the controls. CONCLUSION: Our data suggest that altered lipid profiles in women with PCOS may result from the independent effects of androgen excess and insulin resistance. PMID- 10682454 TI - [Influences of sperm quality and quantity on fertilization, cleavage rates and quality of embryos in in-vitro fertilization]. AB - OBJECTIVE: To examine the influences of the quality and quantity of seminal sperm and inseminational sperm on in-vitro fertilization (IVF) outcomes. METHODS: chi square tests were used to analyze the impacts of both semen and inseminated sperm concentrations and percentages of different grade motile sperm on the fertilization rates in 481 matured and 273 immatured oocytes and cleavage rate percentage of embryo with low quality in embryos obtained from 92 IVF-embryo transfer cycles. RESULTS: The fertilization rates of both kinds of oocytes and cleavage rate of fertilizaed matured oocyte significantly decreased (P < 0.05) as the concentrations and percentages of grade a and b sperms in semen reduced. The rates of embryos with abnormal morphology were significantly dropped when the above sperm parameters increased (P < 0.05). Meanwhile, it was also found that the concentration and motility of inseminated sperm could also affect the above three IVF parameters significantly (P < 0.05). CONCLUSION: Highly active semen and inseminated sperm play important roles not only in fertilization, but also in cleavage and embryo development during IVF procedure. PMID- 10682455 TI - [Comparative study of tetracycline-estrone and estrone effects on bone histomorphometric parameters in ovariectomized rats]. AB - OBJECTIVE: To compare the effects of tetracycline-estrone (TE) and estrone (E) on bone histomorphometric parameters of femoral distal diaphysis in ovariectomized (OVX) rats. METHODS: Twenty female rats were randomly allocated into four groups, 5 rats in each: tetracycline-estrone (TE), estrone (E), OVX and sham operation (S). The OVX rats were used as a model for osteoporosis. After being fed with TE or E for 13 weeks, all rats were sacrificed. The effects of TE and E on bone microarchitecture and dynamics were studied by bone histomorphometry. RESULTS: The histomorphometric data showed that the connectivity of trabecular bone in TE and E groups was significantly improved in comparision with that in S group (P < 0.05). The dynamic data indicated that in TE and E group, the tetracycline labelled and osteoid surfaces were remarkably increased in comparision with those in OVX group, especially the data in TE group were significantly higher than those in E group and other two groups (P < 0.05). CONCLUSIONS: The connectivity of trabecular bone could be significantly improved by both TE and E. The activation frequency seemed to be higher in TE group than that in E group. PMID- 10682456 TI - [Treatment of 129 patients with advanced and recurrent malignant ovarian germ cell tumor]. AB - OBJECTIVE: To investigate the improved treatment for advanced malignant germ cell tumors of the ovary. METHODS: 129 patients with advanced and recurrent germ cell tumor of the ovary treated from 1958 to 1993 were retrospectively analyzed. The results of malignant nondysgerminomatous germ cell tumor of the ovary treated from 1958-1983 were compared with those treated with the supplement of VAC (Vincristine + ActinomycinD + Cyclophosphamide), PVB (Cisplatin + Vinblastine + Bleomycin) or BEP (Bleomycin + Etoposide + Cisplatin) combined chemotherapy from 1984-1993. RESULTS: The overall 5-year survival rate was 30% (39/129) of advanced and recurrent germ cell tumor of the ovary. The 5-year survival rates of dysgerminoma and nondysgerminomatous germ cell tumor of the ovary were 68% (19/28) and 20%(20/101), respectively. The 5-year survival rate of patients with malignant nondysgerminomatous germ cell tumor of the ovary was raised from 3% (1/40) to 42% (15/36) in the recent years. The institution of combination chemotherapy is critical for the improvement with BEP regimen as the best among the three. CONCLUSIONS: The endodermal sinus tumor of the ovary stands first in the incidence of malignant germ cell tumors of the ovary in China. To improve the prognosis of advanced malignant germ cell tumor of the ovary, BEP regimen is the most effective at present. PMID- 10682457 TI - [Effects of mifepristone on gene expression of epidermal growth factor in human uterine leiomyoma]. AB - OBJECTIVES: To study the effects of mifepristone on epidermal growth factor gene expression in human uterine leiomyoma. METHODS: 20 patients with leiomyoma were divided into two groups. One was control group who underwent hysterectomy because of leiomyoma, the other was experimental group who underwent hysterectomy after pretreatment with mifepristone 10 mg/daily for 3 months. Epidermal growth factor (EGF) mRNA was semiquantified in samples of leiomyoma and adjacent normal myometrium from patients untreated in different phases of menostrual cycle and from those treated with mifepristone. Semiquantitative reverse transcriptase polymerase chain reaction, using beta-actin as internal standard, was applied to determine levels of EGF mRNA. RESULTS: Leiomyoma untreated with mifepristone had significantly greater amounts of EGF mRNA than adjacent normal myometrium of uterus only in the luteal phase, but not in the follicular phase of cycle. Similarly, leiomyoma untreated with mifepristone also had significantly larger amount of EGF mRNA than treated leiomyoma in the luteal phase of the cycle, whereas, no difference in the follicular phase of the cycle. CONCLUSION: These findings suggested that: (1) EGF mRNA levels in leiomyoma were increased only in the luteal phase, therefore, maybe mainly controlled by progesterone; (2) mifepristone inhibited EGF gene expression in leiomyoma. This may be one of regression mechanism of uterine leiomyoma in response to the antiprogesterone mifepristone. PMID- 10682458 TI - [The influence of immediate postpartum insertion of GyneFix PP-intrauterine devices on puerperal period]. AB - OBJECTIVE: To investigate the effects of immediate postpartum insertion of GyneFix PP-intrauterine devices (IUD) on postpartum hemorrhage, bloody lochia and postpartum endometritis. METHODS: 126 women had GyneFix PP-IUD inserted immediately postpartum (IUD group) and another 118 cases with no IUD insertion were served as control. Blood loss during delivery, 2 and 24 hours after delivery were determined, the time of bloody lochial lasted and the incidence of postpartum endomeritis were recorded. RESULTS: There were no significant differences in the blood loss during and after delivery, and the incidence of endometritis between the 2 groups, but the bloody lochia was lasted significantly longer in the IUD group (P < 0.05). CONCLUSION: This results suggested that the immediate postpartum insertion of the GyneFix PP-IUD is a safe and good postpartum contraceptive method, and the cause of prolonging bloody lochia awaits for further study. PMID- 10682459 TI - [Clinical uses of misoprostol in obstetrics and gynecology]. PMID- 10682460 TI - [Clinical uses of goserelin in gynecologic diseases and its safety]. PMID- 10682461 TI - [The changes of maternal and umbilical serum nitric oxide in patients with pregnancy induced hypertension]. AB - OBJECTIVE: To determine whether levels of maternal and umbilical serum nitric oxide (NO) were changed in patients with pregnancy induced hypertension (PIH). METHODS: 40 women with PIH, 40 normal late trimester pregnant women and 20 nonpregnant women were studied. Maternal venous blood were collected from all the cases and umbilical venous blood were collected from 28 of the first two groups respectively. Serum NO2-/NO3-, the endproducts of NO, were measured with the Griess reaction after reduction with nitrate reductase. RESULTS: The mean NO2 /NO3- in maternal venous serum were 23.30 +/- 5.60 mumol/L in nonpregnant group, 26.42 +/- 4.54 mumol/L in normal pregnant group and 32.58 +/- 5.06 mumol/L in PIH group. Umbilical serum NO2-/NO3- were 12.26 +/- 4.91 mumol/L in normotensive groups and 14.64 +/- 3.90 mumol/L in PIH group. Maternal serum NO2-/NO3- in nonpregnant group were lower than that both in normal pregnant group (P < 0.05) and in PIH group (P < 0.01). Compared with the normotensive group, maternal serum NO2-/NO3- in PIH group was significantly higher (P < 0.01), while significantly higher serum NO2-/NO3- were also found in umbilical venous blood in PIH group (P < 0.05). There were no correlations between serum NO2-/NO3- level and blood pressure or birth weight. The total nitrite levels in fetal circulation was lower than that in maternal circulation in both pregnant groups (P < 0.01). CONCLUSION: Total nitrites are increased both in maternal and fetal circulation in PIH. The result indicates that the increase of NO may be secondary compensative reaction in PIH. PMID- 10682462 TI - [Study on the activity of platelets protein kinase C in patients with pregnancy induced hypertension]. AB - OBJECTIVE: To investigate the relationship between the activity of protein kinase C (PKC) in plateletes and patients with pregnancy induced hypertension (PIH). METHOD: Activities of PKC in membrance of platelets, which came from 18 PIH patients(PIH group), 20 normal pregnant women (normal pregnant group) and 20 healthy women (control group), were measured by substrate protein phosphoylation method. RESULTS: Activities of PKC in the membrane and cytosolic of platelets from normal pregnancie are lower than those from control group significantly (P < 0.01). Activities of PKC in the membrane and cytosolic of platelets from PIH group are higher than those from normal pregnancie (P < 0.05). Activity of PKC in membrane of platelets from mild and moderate PIH group is higher than that from normal pregnancie (P < 0.01). However, there is no difference between the activities of PKC in cytosolic of platelets from the two groups (P > 0.05). Activities of PKC in membrane and cytosolic of platelets from severe PIH were higher than those from normal pregnancie significantly (P < 0.01, P < 0.05). We also found that the activity of PKC in cytosolic of platelets from severe PIH group was significantly higher than that from mild and PIH cases, though there was no significantly difference in the activity of PKC in membrane of platelets between the two groups. CONCLUSION: Activity of PKC in platelets relates to the occurrence and development of PIH syndrome. PMID- 10682463 TI - [Expectant treatment of placenta previa with ritodrine]. AB - OBJECTIVE: To investigate the effectiveness of the expectant treatment in placenta previa with adrenergic agonist ritodrine. METHODS: 50 women with placenta previa of preterm labor were randomly assigned into two groups. 26 patients treated with magnesium sulfate were served as control group. 24 patients were enrolled in the study group, receiving ritodrine 100 mg in 5% glucose 500 ml intravenous. The drip speed was started at 8 drips per minute routinely, then adjusted according to the treatment response, and oral ritodrine was used after vaginal bleeding and uterine contraction disappeared 12 hours. If contraction reappeared, the i.v. infusion would be restarted. RESULTS: The study group prolonged the gestational period to an average of 28.24 days and increased the birth weight of newborn to an average of 2,913.68 g (P < 0.05). CONCLUSION: Ritodrine is highly effective and safe for expectant treatment of placenta previa. PMID- 10682464 TI - [Clinical experience with iron supplementation in pregnancy]. AB - OBJECTIVE: To study the efficacy of iron supplementation during pregnancy and its influences on the outcome of pregnancy. METHODS: A total of 369 pregnant women were enrolled in this study. According to the hemoglbin levels at recruitment, there were 2 groups: preventive (Hb > or = 110 g/L) and treatment (Hb < 110 g/L) groups. In the preventive group, women entered the study from 20-24 gestational weeks and were randomly assigned to materna treatment (n = 96) who took materna 1 tablet daily or control group (n = 95) who took no other supplementation. In the treatment groups, women less than 36 gestational weeks were accepted. They were randomly divided as materna (n = 93) 1 tablet/d, ferrous sulfate 0.3 g tid/d (n = 50) or Ferroids 1 tablet/d (n = 35) groups. Both Hb and serum ferritin concentrations were determined at admission and immediately after delivery. In some cases serum ferritin in the umbilical vein were measured as well. Hemoglobin levels were examined every 4 weeks during the observational period. RESULTS: In the preventive groups, maternal serum ferritin levels after materna treatment were significantly higher than that before treatment and the control group (P < 0.05). As for the anemia women, compared the serum ferritin concentration, materna treatment had significantly higher levels than that at admission (P < 0.05), and also higher than that in the ferrous sulfate or ferroids tratment groups (P < 0.05). The s-ferritin in the umbilical vein had no correlation with the paired maternal levels. There were no significant differences in the pregnancy outcomes among all the groups. CONCLUSION: Materna can increase the iron storage and effectively improve the iron deficiency during pregnancy, and has no impact on the prgnancy outcomes. PMID- 10682465 TI - [Color Doppler monitoring the utero-placental-fetal circulation variety of normal pregnancy and intrauterine growth retardation]. AB - OBJECTIVE: To study the utero-placental-fetal circulation (UPFC) in normal pregnancy and intrauterine growth retardation (IUGR) cases. METHODS: Color doppler ultrasound was used to detect UPFC in 150 second and third trimester pregnant women, of which 89 cases were normal pregnancy and 58 cases were IUGR. 3 cases were IUGR with chronic renal failure. Hemodynamical value of the umbilical artery (UmA), umbilical vein (UmV) and uterine artery (UtA) were examined directly. The indices included time average velocity (TAV), pulsatility index (PI), resistance index (RI), systolic/diastolic (S/D) ratio, blood flow volume (Q). The maternal serum estriol (E3), human placental lactogen (HPL) and plasma thromboxane B2 (TXB2)/6-keto-PGF1 alpha (6-KP) were measured simultaneously. RESULTS: The result shows that in normal pregnancy group UPFC is abundant gradually with increasing gestational age. In IUGR group 92.53% of cases showed that TAV and Q of UmA, UmV markedly decreased and PI, RI and S/D ratio of UmA elevated at 20 weeks of gestation. There were significant difference between the two groups, maternal serum E3, HPL level in IUGR group were significantly lower than that of the normal pregnancy group, 6-KP level reduced, and TXB2/6-KP ratio significantly increased. CONCLUSION: Using color doppler ultrasound examining hemodynamical changes of UmA, UmV and UtA could observe UPFC function directly. It is one of the best method to early diagnose and predict the prognosis of IUGR. PMID- 10682466 TI - [Morphometric analysis of gap junctions of the cell membrane in human uterine smooth muscle at term]. AB - OBJECTIVE: To study the changes of the gap junctions (GJS) in the uterine smooth muscle cells at various stages of labor, and its relationship with the onset of labor. METHODS: 18 women (38 to 41 gestational weeks) were divided into three groups: not-in-labor, pre-labor, active in labor and six cases in each group. The uterine myometrium tissues were observed by transmission electron microscopy (TEM) and lanthanum tracing method. RESULTS: The lengths and areas of GJS in uterine corpus and lower segment in the active in labor group were significantly larger than that of the other 2 groups, but there was no significant difference between the not-in-labor and pre-labor groups. The area but not the lengths of GJS in corpus was significant larger than that in the lower segment in the active in labor group. CONCLUSION: The great increase of length and areas of GJS in all parts of the uterine smooth muscle was closely correlated with onset of labor. PMID- 10682467 TI - [Cervical ripening score by transperineal ultrasonography and its predictive effect for induction of labor by prostaglandin E2]. AB - OBJECTIVE: To establish a cervical ripening score by transperineal ultrasonography for induction of labor by Prostaglandin E2, and evaluate its predictive effect. METHODS: 105 primiparae underwent cervical assessment by digital and transperineal ultrasonography examination. The five indices of cervical status, other clinical parameters and latency period (from induction to the onset of labor) were analysed by Cox multivariate model. RESULTS: Transperineal ultrasonography was simple, visualable satisfactorily, noninvasive and less uncomfortable. The width of cervical internal os, the length of the canal and position of the presenting part were the main factors that influenced the latency period (P < 0.05). Primiparae with score > or = -4.5 were easy to initiate the onset of labor within 12 hours (P < 0.001). The predictive results were in good agreement with clinical outcomes, and had less misdiagnosis (the Kappa value, specificity and the positive predictive value were 0.7409, 0.7917 and 0.9254, respectively), and were better than the Bishop score (the corresponding value were 0.5680, 0.6667 and 0.8806, respectively). CONCLUSIONS: Transperineal ultrasonography is safe and effective. In predicting the effect on induction of labor of prostaglandin E2 in primiparae, the transperineal ultrasonographic cervical ripening score is a valuable method. PMID- 10682468 TI - [Experimental study on the role of insulin-like growth factor-I in ovulation induction]. AB - OBJECTIVE: To investigate the possibility of insulin-like growth factor-I (IGF-I) as an adjunctive factor for ovulation induction. METHODS: The model of anovulatory mice was established by single injection of testosterone propionate. Thirty anovulatory mice were divided into 3 groups, each contained 10 mice. IGF-I 1 microgram + human menopausal gonadotropin (hMG) 5 IU, IGF-I 2 micrograms or hMG 10 IU alone was injected intraperitoneally to each group respectively. Cyclic changes of vaginal smear and numbers of oocytes in fallopian tubes were used to assess the effect. RESULTS: Vaginal smears of 8 mice in IGF-I + hMG group, 6 mice in hMG group and 2 mice in IGF-I group returned to normal cyclic pattern. The differences between IGF-I + hMG group and IGF-I or hMG groups were significant (P < 0.01 and P < 0.05), but the difference between IGF-I group and hMG group was not significant (P > 0.05). The average numbers of oocytes within fallopian tubes were 12.3 in IGF-I + hMG group and 9.2 in hMG group (P < 0.05). In IGF-I group there were no oocytes in the fallopian tubes of two mice whose vaginal smears returned to normal. CONCLUSION: IGF-I may be a potential agonist agent for ovulation induction. PMID- 10682469 TI - [Prevalence of premenstrual syndrome in reproductive women and its influential factors]. AB - OBJECTIVE: To investigate the prevalence of premenstrual syndrome and its influential factors in reproductive women. METHOD: A questionnaire was used in 454 reproductive women aged 15-49 in Beijing. RESULTS: The prevalence of premenstrual syndrome in these women was 30.4%, among which 61.6% was mild, 34.1% moderate, and 4.3% severe. The order of frequency of the symptoms occurring in premenstrual syndrome was irritation, depression, anxiety, diarrhea, lack of concentration and hypersomnia. Women with higher cultural level, greater stress in life (work or study), dysmenorrhea, dislike menstruation and those with depression symptom had a higher incidence than those with lower cultural level, no stress in life, no dysmenorrhea, no dislike toward menstruation and no depression symptom (P < 0.05). CONCLUSIONS: Premenstrual syndrome is a common disease in reproductive women. Clinical doctors should pay attention and adopt comprehensive measures in order to reduce its incidence and improve the quality of life especially for women with high risk factors. PMID- 10682470 TI - [The dose-response of cell proliferation and secretion of CA125 II antigen to follicle stimulating hormone in ovarian cancer cell lines]. AB - OBJECTIVE: To study the dose-response of the proliferation and secretion of CA125 II antigen with follicle stimulating hormone (FSH) in epithelial ovarian cancer cell lines. METHODS: Two different kinds of ovarian cancer cell lines originated from different pathologic types-AO and 3AO were studied. Cancer cells were cultured in medium containing FSH in different concentrations. The proliferation rate of cells was detected by using the 3H-TdR intrusion technique. The accordant titer of CA125 II antigen in the culture medium was measured by enzyme linked immunoadsorbent assay technique. RESULTS: Proliferation rate of cancer cells (AO and 3AO) was increased apparently as increasing in the concentration of FSH in the culture medium. A positive correlation was found between them (P < 0.05). Proliferation rate of AO cells positively correlated with secreting quantity of CA125 II antigen (P < 0.05), however, no such correlation was found in 3AO cells. CONCLUSION: It suggested that FSH can stimulate the proliferation of epithelial ovarian cancer cell at high concentration and there is a accordant dose-response relationship between them. The secreting pattern of the CA125 II antigen varied by the pathological classification of the ovarian cancer. PMID- 10682471 TI - [Effect of mifepristone on the expression of progesterone receptor messenger RNA and protein in uterine leiomyomata]. AB - OBJECTIVE: To determine the expression of progesterone receptor (PR) mRNA and PR protein levels in the myometrium and leiomyomata from untreated and mifepristone pretreated women with leiomyoma and to examine the mechanism of mifepristone treatment on uterine leiomyomata. METHODS: Expression of PR mRNA and PR protein were determined by Northern blot and HAP of single-dose saturated analysis in myometrium and leiomyomata (center and marginal area) from 27 untreated and 6 mifepristone pretreated women with leiomyomata. RESULTS: PR mRNA abundance and PR protein levels in both myomatous center and marginal area were significantly greater than those in corporal myometrium (P < 0.01) in both follicular and luteal phases, but similar between myomatous center and marginal area (P > 0.05). 6 cases pretreated with mifepristone 25 mg/day for 3 months were operated, all but one patient displayed a decrease in leiomyomata volume. PR mRNA abundance in both myometruim and leiomyomata (center and marginal area) was significantly decreased in 4 patients continuing mifepristone treatment before the operation but not in the other 2 patients stopping RU486 1 month before operation. PR protein levels in these tissues showed significant decrease in all 6 cases. CONCLUSION: There are overexpression of PRmRNA and PR protein in leiomyomata. One of the mechanism of mifepristone action on decreasing leiomyomata volume may be related to suppression on expression of PR gene. It seems that suppression on transcription of PR gene is reversible, but on translation of PR gene may maintain in a relatively longer period. PMID- 10682472 TI - [The morphometric study of endometrial spiral arterioles before and after insertion of gamma CuI and TCu 220C intrauterine devices]. AB - OBJECTIVE: To investigate the morphologic changes of endometrial spiral arterioles and its relationship with bleeding pattern after insertion of gamma shape copper indomethacin-medicated (gamma CuI) and T-shape copper (TCu 220C) intrauterine devices (IUD). METHODS: Endometrium specimens of late secretory phase were obtained from fertile age women: 10 from preinsertion, 10 obtained after insertion of TCu 220C IUD, and 9 obtained after insertion of gamma CuI IUD. Samples were sectioned serially and morphometric analysis of endometrial spiral arterioles was performed under light microscope. RESULTS: The average cross section area (Area), maximum diameter (Dmax) and minimum diameter (Dmin) of spiral arterioles in both spongeous and dense layers of endometrium increased significantly after insertion of TCu 220C IUD. After insertion of gamma CuI IUD, the Area and Dmax increased in dense layer only, though less obviously than that occurred in TCu 220C group. However, the Dmin increased more obviously in both spongeous and dense layers than after insertion of TCu 220C IUD, implying that the shape of spiral arterioles was more regular in gamma CuI group. CONCLUSION: gamma CuI IUD has less effects on the morphological changes of endometrial spiral arterioles, and this may relate to its indomethacin-contained which causes less bleeding. PMID- 10682473 TI - [Human immunodeficiency virus infection among gynecology and obstetrics patients]. PMID- 10682474 TI - [Gestational diabetes and fetal macrosomia]. PMID- 10682475 TI - [Clinical uses of maternal serum markers in the prenatal diagnosis]. PMID- 10682476 TI - [Human cytomegalovirus infection and congenital malformation]. AB - OBJECTIVE: To study the relationship between intrauterine cytomegalovirus (HCMV) infection and congenital malformation, and to determine the distribution of tissues infected. METHODS: Autopsy samples of 41 infants with congenital malformation and 19 infants with normal appearances were studied. Using polymerase chain reaction (PCR) technique the paraffin embedded specimens of main organs were examined for HCMV infection. In-situ hybridization (ISH) was performed in some of the PCR positive tissues in order to define the distribution of HCMV DNA. RESULTS: 19 of the 41 infants (46.34%) with congenital defects were HCMV positive, while 1 in 19 (5.26%) were positive in the control group, and there was significant difference between the 2 groups (P < 0.05), 20.46% (35/171) of the fetal organ samples were HCMV DNA positive in the malformation group, but only 1 out of 78 samples (1.28%%) was positive in the pulmonary tissue of the control group. More malformations of the digestive system were presented in HCMV infected babies but no statistical significant difference when compared with other systems. Brain tissue had the highest HCMV infection rates (41.37%, 12/29), which was significantly higher than other organs. By ISH technique HCMV DNA was found only in 6 out of 17 PCR positive samples, and they were located at neurons, neurogliocytes, epithelium and interstitial cells of the kidney, and epithelial cells of pulmonary alveolar. CONCLUSION: There are strong correlation between HCMV infection and congenital malformation, and brain is more susceptible to HCMV. By combining PCR and ISH, both sensitivity and distribution of HCMV could be obtained. PMID- 10682477 TI - [Detection of human cytomegalovirus infection in pregnant women and their fetuses by nested polymerase chain reaction]. AB - OBJECTIVES: To evaluate the applicability of nested polymerase chain reaction (PCR) and restriction endonucleases analyses (REA) for the diagnosis of human cytomegalovirus (HCMV) in pregnant women and their fetuses. METHODS: Nested PCR and REA, virus isolation, anti-HCMV-IgM and anti-HCMV-IgA were used for detection of HCMV in peripheral blood of pregnant women, umbilical blood and tissues of stillbirths. RESULTS: Among 367 pregnant women, the HCMV infections rate in the first, second and third trimester of gestation were 8.6%, 1.6% and 7.0%, respectively. The nested PCR had significantly higher detective rate (4.9%) than that of virus isolation (3.0%, P < 0.025). Among 6 samples of maternal HCMV DNA positive blood, 3 matched umbilical samples were HCMV DNA positive as well. Of these 3 cases, HCMV intrauterine infection was considered. HCMV was detected with all measurements in both maternal and umbilical blood except in 1 paired samples specific-IgM were negative. HCMV DNA was found in the lung tissue of one of the 28 stillbirds. CONCLUSION: The diagnostic specificity and sensitivity of HCMV can be raised by nested PCR. PMID- 10682479 TI - [Weight gain pattern in normal pregnant women]. AB - OBJECTIVE: The purpose of this study was to determine the maternal weight gain pattern of normal pregnant women. METHODS: One thousand five hundred and sixty two (1,562) women, with 11,059 records, received prenatal care and had their babies delivered at the Second Hospital of West China University of Medical Sciences during the period of Jan. 1-Dec. 31, 1995. All the women without complications and with their babies weighted between 2,500-3,999 g were included, and data was analyzed. RESULTS: The average age was 26.49 +/- 3.02 and height 158.69 +/- 4.85 cm. The women were divided into three categories according to their prepregnant body mass index (BMI): underweight (< 16.75), normal weight (16.75-23.71) and overweight (> 23.71). The BMIs of each categories increased as gestational week approached to the term, with total BMI increase as 8.07, 5.37, 3.82 for underweight, normal weight and overweight, respectively. CONCLUSION: BMI, calculated with weight and height, is more accurate to assess maternal weight gain than weight alone do, and above results and accompanied charts could be used to monitor maternal weight gain as a reference. PMID- 10682478 TI - [Prenatal gene diagnosis of alpha-thalassemias]. AB - OBJECTIVE: To study the value of polymerase chain reaction (PCR) in prenatal diagnosis of alpha thalassemias. METHODS: Amniotic fluid prenatal gene diagnosis with polymerase chain reaction was carried out on eleven fetuses whose parents are both heterozygotes with alpha-globin gene deficiency. A DNA fragment of 224bp in the production means normal alpha-globin gene sequence, while a 630bp fragment indicated the alpha-globin gene deficiency. Both 224bp and 630bp fragments in the same sample means heterozygote. RESULTS: Three of the 11 fetuses (one pregnancy was twin) were with normal alpha-globin gene sequence, while 4 were homozygotes and the other 4 were heterozygotes. For the 3 fetuses with ascitic fluid under ultrasound examination, 2 were homozygotes and the other one was heterozygote by gene diagnosis. Two of the 4 homozygotes from induced abortion were typical Bart's syndrome, one was edema in the whole body and the other one with short limbs and abdominal hernia. CONCLUSION: The method of PCR in prenatal diagnoses for detection of alpha-thalassemias is simple, accurate and rapid. PMID- 10682480 TI - [The evaluation of umbilical arterial Doppler spectrum fractal characterization in obstetrics]. AB - OBJECTIVE: To evaluate the clinical value of fetal umbilical arterial Doppler spectrum fractal. METHODS: 104 cases with 22-41 gestational weeks were included, and divided into two groups: normal pregnant group, 59 cases; abnormal pregnant group, 45 cases. Umbilical arterial Doppler spectrum were obtained by color Doppler ultrasound, then Doppler signal were transformed into computer through sound-frequency, and fractal were calculated. RESULTS: The fractal was 1.83 +/- 0.03 in normal pregnant group. The frequency of abnormal fractal was significantly higher in abnormal pregnant group than that in normal pregnant group (P < 0.001). Compared with the ratio of peak systolic to lowest diastolic flow velocity (S/D), the fractal was more sensitive than S/D ratio in abnormal pregnancy (P < 0.05). There were significant correlation between fractal and gestational weeks (r = 0.266). CONCLUSION: Fractal responses to complexity of umbilical arterial blood flow, and it is a better parameter than S/D ratio in monitoring abnormal umbilical arterial blood flow. PMID- 10682481 TI - [The changes of serum nitric oxide level in patients with intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To study the relationship of serum nitric oxide (NO) level with pathophysiological changes in intrahepatic cholestasis of pregnancy (ICP). METHODS: 49 patients with ICP and 26 healthy women in their late pregnancy were studied. The serum NO levels were determined with Greiss reaction by measuring oxidation products of NO. RESULTS: The serum NO concentration in patients with ICP decreased significantly when compared with that of healthy pregnant women. The serum NO level in patients with ICP was related to the time of persistent pruritus, the serum concentration of bilirubin and glutamic-pyruvic transaminase (GPR). There is no correlation between serum NO and cholylglycine. In the ICP group the average gestational age was shorter than that of normal pregnancy and the weight of newborn was also lower. CONCLUSIONS: The decline of serum NO may play an important role in the pathophysiological changes in ICP and may cause the intrauterine growth retardation, premature delivery and fetal hypoxia. PMID- 10682482 TI - [Assessment of fetal maturation by epithelial growth factor in serum of pregnant women]. AB - OBJECTIVE: To study the concentrations of human epithelial growth factor (EGF), progesterone (P) in serum, and EGF, P, amylase (Ams), creatinine (Cr) and total bilirubin (TB) in amniotic fluid at different trimester normal pregnancy. METHODS: The concentrations of EGF, P in serum of 181 cases of pregnant women (first trimester 35, midtrimester 69, late trimester 77) and the concentrations of EGF, P, Ams, Cr, TB in amniotic fluid of 87 cases (mid-trimester 44, late trimester 43) were determined. At the same time, the concentrations of EGF, P in serum of umbilical veins and arteries from 23 full term neonates were determined as well. The concentrations of EGF and P were measured by radioimmunoassay and Ams, Cr, TB in amniotic fluid by PA110 autobiochemistry analyzer. RESULTS: (1) The concentrations of EGF, P in serum increased as pregnancy advanced. (2) There were significant correlations between EGF level and Ams, Cr levels in amniotic fluid. After 32 gestational weeks, the fetal maturity rate was 70.59% when EGF was > or = 4.5 micrograms/L. There were significant positive correlations between EGF in maternal serum and in amniotic fluid, (3) The maternal serum EGF concentration was significantly higher than that in the umbilical vessels, and there was positive correlations between them. CONCLUSION: The change of maternal serum concentration could be used to determine the fetal maturation. PMID- 10682483 TI - [Functional states of pituitary-ovary, adrenal and thyroid axes in women with polycystic ovarian syndrome]. AB - OBJECTIVE: To assess the role of the functional states of pituitary-ovary, adrenal and thyroid axes in the pathophysiology of polycystic ovarian syndrome (PCOS). METHODS: The stimulation tests of above-mentioned three endocrine axes by luteinizing hormone releasing hormone (LRH, 100 micrograms), adrenocorticotrophin (ACTH, 250 micrograms) and thyroid releasing hormoune (TRH, 500 micrograms), respectively, were performed in each subjects of two PCOS groups [luteinizing hormone/follicle stimulating hormone (LH/FSH) > or = 3, Group 1, n = 15; LH/FSH < 3, Group 2, n = 15] and the control (n = 20). Endocrine indices of corresponding target glands were evaluated by radioimmunoassay (RIA). RESULTS: After LRH administration, the amplitude of LH responses in three groups were as follows: Group 1 > Group 2 > control while mean testosterone levels were elevated to a similar extent in two PCOS groups and remained unchanged in the control. Following ACTH stimulation, PCOS subjects, especially of Group 2 with obvious insulin resistance, showed higher responses of cortisol, dehydroepiandrosterone sulfate and testosterone as compared with the controls. However, during TRH testing, exaggerated thyroid-stimulating hormone and prolactin responses in two PCOS groups and blunted free thyroxine response at two hour point in Group 1 were observed, as compared with the control. CONCLUSION: It appears that ovarian androgen excess in women with PCOS is mainly LH-dependent in Group 1 and insulin dependent in Group 2. Enhanced adrenal activity may contribute to both hyperandrogenism and insulin resistance in this syndrome, and subclinical hypothyroidism may exist in affected subjects, especially of Group 1. PMID- 10682484 TI - [The effect of sex steroids on human ovarian granulosa cell apoptosis]. AB - OBJECTIVES: To study the molecular mechanism of human follicular atresia by exploring the effects of estrogen, androgen on human granulosa cell (GC) apoptosis. METHODS: Ovarian GC DNA fragments of both atretic and developing follicles before and after estrogen (1 microgram/ml), androgen (1 microgram/ml) treatments were analysed by agarose gel electrophoresis. The expression of bcl-2 mRNA was also determined by northern blotting in GC of developing follicle after sex steroids addition. RESULTS: Internucleosomal DNA cleavage occurred in granulosa cells in atretic follicles and after androgen treatment. In granulosa cells of developing follicles, estrogen treatment increased the expression of bcl 2 mRNA by 45%, while androgen treatment downregulated the expression of bcl-2 mRNA by 35%. CONCLUSION: Androgens increase apoptotic DNA fragmentation, and decrease the expression of bcl-2 mRNA in GC of developing follicles, while estrogens produce the opposite effect. Therefore, expression of bcl-2 may be involved in the mechanism of follicular development and atresia. PMID- 10682485 TI - [Patterns of cisplatin induced apoptosis in ovarian cancer cell line COC1]. AB - OBJECTIVE: To elucidate the patterns of chemotherapeutic drugs induced apoptosis and its role in cancer chemotherapy. METHODS: Apoptosis in cultured ovarian cancer cell line COC1 induced by ciplatin was investigated in vitro by applying cytohistochemistry stain, florescence stain and electron microscopy scanning combining with DNA gel electrophoresis and flow cytometry analysis. RESULTS: After exposure to cisplatin, COC1 cells manifested typical apoptotic morphological features. Apoptosis persisted throughout 30 hours following addition of cisplatin and augmentated gradually. The rate of apoptosis was enhanced within tested dose range in a dose dependent pattern. At the point of 12 hour cisplatin exposure, the necrosis rate was only 12.4% while the apoptotic rate accounted for 30.8% already. CONCLUSIONS: Chemotherapeutic drugs induced apoptosis is one of the primary mechanism of anti-cancer chemotherapy. PMID- 10682486 TI - [The relationship between the pathological classifications and molecular genetics of hydatidiform moles]. AB - OBJECTIVE: To study the relationship between the pathological classification and molecular genetics of hydatidiform mole. METHOD: 32 cases of hydatidiform mole were analyzed by DNA restriction fragment length polymorphism (RFLP, hybridized with the probe 33.15). RESULTS: DNA from only paternal origin was found in 21, and from both parents in 11. In the formeron the basis of pathological characteristics, the complete hydatidiform mole (CHM) and the partial hydatidiform mole (PHM) were 16 (76%, 16/21) and 5 (24%, 5/21) respectively, and in the later CHM and PHM were 5 (45%, 5/11) and 6 (55%, 6/11), respectively. CONCLUSION: There is not much correlation between pathological classification and molecular genetics of hydatidiform mole. PMID- 10682487 TI - [Detection and sequence analysis of the p53 gene mutation in epithelial ovarian cancer]. AB - OBJECTIVE: To find the characteristics of p53 gene mutation in epithelial ovarian cancer and to analyze the relationship between p53 mutation and FIGO stage. METHODS: p53 mutations in exon 5 to 7 were detected by single-strand conformational polymorphism (SSCP) and sequencing technique. RESULTS: 8 of 46 tumor tissues demonstrated a SSCP band shift in the region of the gene. All of them have been characterized to represent DNA alterations by sequencing, including 8 point mutations (6 missence, 1 silent mutation and 1 in intron) and a 1-base pair insertion (introducing a stop codon downstream). Overall, 88.9% of mutation were transitions, and most of them are G-->A transitions (7/8, 87.5%). 62.5% of the mutation were found in 175 and 245 codon. The percentage of the mutation in stage I and stage II was 20.0%, and in stage III and stage IV was 16.7% (P > 0.05). CONCLUSION: The arising of p53 mutations in ovarian cancer is due to spontaneous error in DNA synthesis and repair. Codon 175, 245 are the two mutational hot spots. There is no relationship between the mutation of p53 gene and FIGO stage in epithelial ovarian cancer. PMID- 10682488 TI - [Investigation of intrauterine microbes after intrauterine operation]. AB - OBJECTIVE: To explore the postoperative changes in the cultures of ureaplasma urealyticum (UU), mycoplasma hominis (Mh), L-form bacteria (L-form), anaerobic bacteria (Ana) and chlamydia trochomatis (CT) after intrauterine operation. METHODS: Four groups of patients were set up: group 1, induced abortion; group 2, intrauterine device (IUD) insertion; group 3, penicillin i.m. after IUD insertion; group 4, oral lincomycin after IUD insertion. Intrauterine secretion were aspirated to identify the above microbes before operation and within 1 week of ending of menstrual bleeding for 4 consecutive postoperative cycles. Bacteria carrier was defined as at least one of the 5 microbes detected. RESULTS: No difference was shown in the incidence of bacteria-carrier (IBC) among the 4 groups preoperation. The IBC tended to be the highest in the first menstrual cycle postsurgery in all the 4 groups, then decreased. Compared with preoperation, there were significantly higher IBC in the 3 IUD groups (P < 0.05) except group 1. CONCLUSION: IUD is a major factor for intrauterine microbes existing after operation, and the natural body defense system can help to get rid of the organism by time. Small doses and short period of penicillin or lincomycin administration proved not effective in clearing the intrauterine microbes after IUD insertion. PMID- 10682489 TI - [Predicative factors of long-term occurrence of diabetes in the patients with gestational diabetes]. PMID- 10682490 TI - [New concepts of pathology in serious borderline tumor in ovarian neoplasm]. PMID- 10682491 TI - [Ovarian malignant tumor metastatic to the breast]. PMID- 10682492 TI - [A team of modern epidemiologists are needed in China]. PMID- 10682493 TI - [Epidemiologic study and prevention and control of communicable diseases in army in China: experiences and successes]. PMID- 10682494 TI - [Zeng-Ding phenomenon: further demonstration and studies on its predictive value in epidemic of measles and scarlet fever]. AB - OBJECTIVE: To demonstrate further the existence of Zeng-Ding phenomenon in disease epidemic and to explore the relationship between it and the time series in different kinds of diseases. METHODS: Incidence data of notifiable communicable diseases during 1975 to 1996 were collected. Time series of measles and scarlet fever incidence during 1975 to 1995, 1980 to 1995, 1985 to 1995 and 1990 to 1995 were established. Correlation analysis was conducted between monthly cumulative percentage and predictive ratio of increase to decrease in incidence rate at the best cut-off point. Prediction was studied based on the constructed extrapolation model. RESULTS: Correlation analysis showed that 98.3% (232/236) of the coefficients of correlation were negative (R < 0), indicating further the existence of Zeng-Ding phenomenon in disease epidemic. There was significant difference in coefficients of correlation between the four time series, which accounted for 63.8%, 54.2%, 44.1% and 35.0%, respectively; and for 73.3%, 56.7%, 36.7% and 36.7%, respectively, in measles, and for 53.6%, 51.8%, 51.7% and 33.3%, respectively, in scarlet fever. It showed that Zeng-Ding phenomenon correlated with the time series and the kinds of diseases. Prediction from extrapolation model showed that there was significant difference in predictive agreement between two time series of 1975 to 1995 (65.5%) and 1985 to 1995 (37.0%) with chi 2 of 4.54 and P < 0.05, indicating a trend that predictive agreement increased with prolonging of time series, and a trend that predictive agreement for scarlet fever increased with decreasing of coefficients of correlation. CONCLUSION: Predictive value of the incidence data can be evaluated by their source and length of time series. PMID- 10682495 TI - [Detection of S-gene mutation strain in vertical transmission of HBV and its significance]. AB - OBJECTIVE: To study S-gene mutation of hepatitis B virus (HBV) in its vertical transmission and its significance. METHODS: Nucleotides of S-gene NT451-660 of HBV were sequenced with dideoxy end termination technique in four female and six male carriers without HBV markers in their spouses and in their intrauterine infected fetuses. RESULTS: It was showed that homology of HBV nucleotide and amino acid sequences in the mothers, fathers and their fetuses was very high. Mutation at the sites 491, 494, 530, 546 and 581 of S-gene resulted in amino acid substitution at the sites 113, 114, 126, 131 and 143, respectively. Mutations at the sites 126 were detected in two pairs of mother or father and her or his fetuses and mutations at the sites 131 in four fetuses, respectively, including combined mutation at the site 143 in two fetuses. CONCLUSION: Strains with S-gene mutation, mainly at the sites 126, 131 and 143, could be found in HBV vertical transmissions, which could cause failure in HB vaccine immunization. PMID- 10682496 TI - [Case-control studies on risk factors for pulmonary tuberculosis in servicemen of China]. AB - OBJECTIVE: To study the risk factors for pulmonary tuberculosis (PTB) in the servicemen of armed forces and lay a basis for its prevention and control. METHODS: A 1:2 matched case-control study was carried cut in the servicemen, with 86 cases and 153 controls. Data were analyzed with conditional logistic regression with EGRET software. Odds ratios (ORs) were calculated for the relevant factors. RESULTS: Single-variable analysis showed that schooling, history of exposure to PTB, history of Bacille Calmette-Guerin (BCG) vaccination and BCG vaccination scar all associated with the incidence of PTB, with ORs of 2.19, 2.03, 0.38 and 0.54, respectively, and P < 0.05. Multiple logistic regression analysis showed that BCG vaccination scar, history of exposure to PTB and schooling all entered the final regression model, with ORs of 0.36, 1.94 and 2.33, respectively. CONCLUSION: It suggested that BCG vaccination could play a role in the prevention of TB in the servicemen, and history of exposure to PTB was a potential risk factor for TB as well. PMID- 10682497 TI - [Seroepidemiological study on Helicobactor pylori infection in rural adult residents]. AB - OBJECTIVE: To study the prevalence of Helicobactor pylori (Hp) infection in rural adult residents of China and its relation to serum levels of pepsinogen I (PG I), pepsinogen II (PG II) and gastrin (GAS). METHODS: Serum levels of antibodies against Hp were determined with enzyme-liked immunosorbent assay (ELISA) in 1,504 residents aged over 30 in Zanhuang County of Hebei Province, and their serum levels of PG I, PG II and GAS, as well as PG I/PG II ratio, were analyzed quantitatively, with radioimmunoassay (RIA). RESULTS: Positivity for serum Hp antibody accounted for 66.4% of the rural adults in Zanhuang County, without significant sex difference. There was no significant difference in positivity for serum Hp antibody between residents in the high and low prevalent areas of gastric cancer. Serum levels of PG I, PG II and GAS were significantly higher in those with positive serum IgG anti-Hp (62.3 micrograms/L, 15.45 micrograms/L and 74.00 pg/ml, respectively) than those negative (42.1 micrograms/L, 6.40 micrograms/L and 66.00 pg/ml, respectively), all with a P-value less than 0.005, and the ratio of serum PG I to PG II levels was significantly lower in the former (4.0) than that in the latter (6.6), with P < 0.005. CONCLUSION: More than two thirds of the adult residents in Zanhuang County had infected with Hp, which could affect their serum levels of PG and GAS. PMID- 10682498 TI - [Prevalence of HCV and HBV infection in patients with primary hepatocellular carcinoma in Shanxi Province]. AB - OBJECTIVE: To study the prevalence of hepatitis C and B viruses (HCV and HBV) infection in the patients with primary hepatocellular carcinoma (PHC) in Shanxi Province and evaluate their etiologic roles in the pathogenecity of PHC to lay a foundation for the prevention and control of it. METHODS: Ninety-eight patients with PHC and 196 non-PHC controls matched in age and sex were selected from the four hospitals at provincial level in Shanxi Province. Serum antibodies against HCV (anti-HCV), HBs (anti-HBs), HBe (anti-HBe) and HBc (anti-HBc), IgM-antibody against HBc (anti-HBc-IgM), HBsAg and HBeAg were determined for all of them with enzyme-linked immunosorbent assay (ELISA). RESULTS: Positivity for anti-HCV and rate of HBV infection were 8.16% and 63.37%, respectively, in the PHC patients, both significantly higher than those in the controls (P < 0.05), with attributable risk proportions of (ARP) 94%, and 91%, respectively. Multiple conditional logistic regression analysis showed that positive serum anti-HCV, HBsAg and anti-HBc a risk factors for PHC, all entered the regression model, with odds ratios (ORs) of 55.06, 10.18 and 9.85, respectively. Dichotomized contingency table analysis showed that OR for those positive both for anti-HCV and HBsAg was 61.37, significantly higher than that for single positive of either one, with an additive effect. CONCLUSION: It suggested that both HCV and HBV infection were main etiologic factors for PHC in local. HBV also was an important factor in the pathogenecity of PHC, especially in those with both positive for HBsAg and anti-HBc. Coinfection with HBV and HCV had an additive effect on PHC pathogenecity. PMID- 10682499 TI - [Demonstration for periodicity of leptospirosis in Yichang City of Hubei Province during 1960-1997]. AB - OBJECTIVE: To study the periodicity of leptospirosis incidence. METHODS: Data of leptospirosis epidemics in Yichang City collected during the past 38 years from 1960 to 1997 were analyzed with periodic graphics method. RESULTS: Cause-specific incidence rate of leptospirosis was 15.43 per 100,000 in average, with an epidemic interval of 10 years, with statistical significance in periodic vibration (J = 4.179, P < 0.05). CONCLUSION: The results mentioned above provided scientific basis for the effective control and prevention of the disease. PMID- 10682500 TI - [Surveillance of hemorrhagic fever with renal syndrome and studies on its predictive indices in Jiangsu Province]. AB - OBJECTIVE: To study its predictive significance of the direct use of surveillance data of hemorrhagic fever with renal syndrome (HFRS) collected in Jiangsu province, and find an indicator of choice for qualitative prediction. METHODS: A correlation analysis between the HFRS incidence rate and surveillance data collected in the past 12 years since 1986 was applied. RESULTS: There was significant relationship between HFRS incidence rate and indoor density of rattus norvegicus (Rn), proportions of mixed species of rats and Rn with HFRS virus, indices of mixed species of rats and Rn with virus in the spring Relationship between HFRS incidence rate and densities of outdoor mixed species of rats and Apodemus agrarius (Aa), and proportions of mixed species and Aa with virus was all significant in autumn and winter. And, there was significant relationship between HFRS incidence rate during the whole year and annual average density of rats, proportion of rats with virus, index of rats with virus. There was no significant relationship between HFRS incidence rate and human inapparent infection rate. The coefficients of correlation between HFRS incidence rate in the spring and indices of indoor mixed species of rats and Rn with virus were 0.8637 and 0.8295, respectively (P < 0.001). And, those between HFRS incidence rate and indices of outdoor mixed species of rats and Aa were 0.7089 and 0. 7258 in the autumn and winter, respectively (P < 0.01). The coefficients of correlation between HFRS incidence rate in the next spring and indices of outdoor mixed species and Aa with virus in the autumn were 0.7118 and 0.7113, respectively (P < 0.01). The coefficient of correlation between HFRS incidence rate during the whole year and annual average index of rats with virus was 0.9207 (P < 0.001). CONCLUSION: The index of rats with virus was an indicator of choice for qualitative prediction of HFRS, and the density of rats may be the secondary choice. PMID- 10682501 TI - [Cost-benefit analysis for hepatitis A vaccine]. AB - OBJECTIVE: To improve economic benefits of hepatitis A (HA) vaccination and to lay a foundation for formulating an immunization strategy for it. METHODS: Health economics methods were used for analyzing the cost-benefit ratio, balance point of cost-benefit and balance point of antibody level after HA vaccination. RESULTS: The benefit-cost ratio (BCR) for HA vaccine was 2.53 in Jiangxing City of Zhejiang Province with an HA-specific incidence rate of 41.15 per ten thousand. Incidence rate of HA was 16.26 per ten thousand at balance point of cost-benefit of HA vaccine. Cost would be reduced if serum HA antibody was screened before vaccination in the population with more than 50% of seropositive HA antibody. CONCLUSION: It indicated that more economic benefits would be gained if mass HA vaccination strategy was used. Vaccinee of choice was those at ages of 15 to 29 years. HA vaccination after antibody screening in the population aged over 25 years would be more economic than the direct use. PMID- 10682502 TI - [A case report of heterosexual transmission of HIV infection between spouses with persistent condom use]. AB - OBJECTIVE: To report a case of heterosexual transmission of HIV infection between spouses with consistent condom use. METHODS: The first case with HIV infection was detected in the contracted male workers returned from abroad in Qingdao in 1992 and both he and his spouse were given health education for AIDS/HIV prevention and instructions on condom use. And, his wife was followed-up every six months by detecting her serum anti-HIV. RESULTS: The wife of the index case was negative for anti-HIV in the first 12 blood tests before December 1996. But, she was found suspect of positive for anti-HIV an July 1997 and definitely positive for anti-HIV-1 in August 1997 during her 14th test. In the period of 1992 to 1997, they persisted in condom use during intercourse. Recall of the wife showed that she had low fever in May 1997, suggesting at the window phase of her acute HIV infection. It postulated that she infected with HIV curing March to April 1997. CONCLUSION: The case report indicated that incorrect use of condom could not prevent from HIV transmission between spouses. PMID- 10682503 TI - [Quality evaluation of locally-prepared kits for HIV antibody detection in clinical applications]. AB - OBJECTIVE: To evaluate the quality of eight kinds of locally prepared enzyme linked immunosorbent assay (ELISA) kits for HIV antibody (anti-HIV) detection and study their current status in clinical applications. METHODS: Two hundred serum specimens were tested with eight kinds of locally prepared ELISA kits for anti HIV screening, with an imported kit as reference, including 100 specimens with confirmed positive, negative or undetermined anti-HIV and 100 specimens collected from the drug abusers in Xinjiang Region. RESULTS: Anti-HIV could be detected in all the 74 confirmed positive specimens with the imported reference kit, with sensitivity of 100%. But, six to eighteen specimens were false negative detected with local prepared kits, with sensitivities of 81.1%-91.9%, mainly in those collected from the drug abusers with weak positive for anti-HIV. Two of the 107 confirmed negative specimens were false positive by the imported reference kit, with a specificity of 98.2%, and 0-8 specimens were false positive by locally prepared kits, with specificities of 92.5%-100%. CONCLUSION: The sensitivity of locally prepared ELISA kits for anti-HIV screening should be improved further to ensure the safety of blood transfusion and the control of AIDS/HIV. PMID- 10682504 TI - [Experiences and achievements in the control of schistosomiasis in China]. PMID- 10682505 TI - [Experiences in the basic control of sexually transmitted diseases and their recurrences in China]. PMID- 10682506 TI - [Review on the study of Keshan disease in China]. PMID- 10682507 TI - [Risk factors for initiation of drug use among young males in Longchuan, Yunnan]. AB - OBJECTIVE: To identify the risk factors for drug use among young males in Longchuan, Yunnan. METHODS: A non-concurrent cohort study was carried out based on a cross-sectional survey. Demographic, behavioral, and drug-using related information were collected using an anonymous questionnaire. The non-concurrent cohort included the period January 1, 1991 to August 1, 1994. Risk factors were assessed by univariate and multivariate analysis. RESULTS: The annual incidence of drug use increased between 1991 and 1993. Multivariate analysis identified the following risk factors for drug use: being born to Jingpo ethnic group (OR = 1.8, 95% C.I. 1.2-2.5), being divorced/ widowed/separated (OR = 8.9, 95% C.I. 1.8 43.3), smoking cigarettes (OR = 2.4,95% C.I. 1,6-3.8), having had pre-/extra marital sex (OR = 1.5, 95% C.I. 1.1-2.2), having been encouraged by friends to try drugs (OR = 8.8, 95% C.I. 6.1-12.9) and having a family member who used drugs in 1991 (OR = 1.5, 95% C.I. 1.0-2.3). More than six years of education was a protective factor for drug use (OR = 0.6, 95% C.I. 0.4-0.98). The population attributable fractions were 70.8% for being encouraged by friends or others to try drugs, 50% for smoking cigarettes and 24% for being Jingpo ethnic group. CONCLUSION: Results suggested that community based programs to change attitudes towards smoking cigarettes and drug use could dramatically reduce the incidence of new drug users in Longchuan County. We, therefore, recommend a community-based intervention program targeting adolescent. PMID- 10682508 TI - [Logistic regression analysis of female drug abusers' social-psychological factors]. AB - OBJECTIVE: To study the social-psychological health status on female drug abusers. METHODS: Case-control study and logistic regression analysis. RESULTS: Results of simple logistic regression showed that factors with significant ORs were low education level, unemployment, marital status, senseless of life, self killing, horror, madness, depression, insomnia, dizziness, perspiration, evil dream, divorce, setbacks of emotion, smoking, alcohol-abuse, playing truant, runaway from home, fighting, etc. The results of stepwise logistic regression analysis showed that factors entered the regression model were smoking, runaway from home, divorce, setbacks of emotion, marital status, hopeless toward life, level of education, dizziness, etc. CONCLUSION: Most female drug abusers' social psychological status was poorer than controls preceding to their drug abuse, which contributed to the major causes of abuse drugs. PMID- 10682509 TI - [Analysis on the epidemic characterization and trend of AIDS in Fujian Province]. AB - OBJECTIVE: To analysed the epidemic characterization and trend of AIDS in Fujian Province based on data obtained from HIV sero-survallance, HIV-1 subtyping and epidemiological survey. METHODS: Serum samples collected from people in high risk groups and specific subgroups were examined for antibody against human immunodeficiency virus (HIV). Epidemiological investigation was conducted toward people who were sero-positive. Clinical data on patients with AIDS were also collected. Peripheral blood mononuclear cells (PBMCs) obtained from HIV-1 infectious and patients with AIDS were used for DNA amplification between C2-V3 region by PCR. The amplified products were sequenced to identify HIV-1 subtypes. RESULTS: From January 1987 through the end of December 1997, a total number of 569,873 serum samples from high risk groups and specific subgroups including blood donors and people that travelling abroad frequently were examined for HIV antibody Seventy people with HIV-1 sero-positive were identified, accounting for 0.12/1000 of the total detected numbers. Among them, 21 patients with AIDS in which 18 cases had died. HIV/AIDS cases were mainly distributed along the coastal cities as Fuzhou, Quanzhou and Xiamen where economic development was growing rapidly. Most cases were infected abroad and spreading HIV to the country through sexual transmission. Clinical data showed that early syndromes and reversal fever of patients with AIDS were seen but survival time was short. Nucleotide sequencing showed that HIV-1 strains in the province were most subtype E. CONCLUSION: HIV epidemic in the province will mainly be spreading among people with sexually promiscuous behaviors. PMID- 10682510 TI - [A prevalence study on injuries among 2,553 children 7-16 years old]. AB - OBJECTIVE: In order to find out the present situation and cause of injuries among children. METHODS: A cluster sampling study on the conditions of injuries was conducted among 2,553 children 7-16 years old during the period of October 1996 to September 1997 in Shantou City, Guangdong Province. RESULTS: There was a trend that the incidence of injuries increased with age among children. The overall incidence of injuries was 37.96% with schoolboys higher than schoolgirls (P < 0.05). There were 38.1% children who had more than two episodes. Falls took the leading type of incidence among both sexes and all age groups. Among the causes of injuries, playing, motion, riding and walking ranked the consecutive leading 4 places. The places where injuries occurred were mainly at home and then on campus. Self injured was mostly seen followed with hurt by others (classmate, sibling, et al). Medium and serious injuries took up 8% with a disability rate of 121.4/100,000. CONCLUSION: Some preventive measures were preliminarily suggested. PMID- 10682511 TI - [Progress of speeding up measles control strategies in Anhui]. AB - OBJECTIVE: To speed up measles control program. METHODS: Catching-up immunization campaigns on measles have been conducted among the children aged 1-6 years in three prefectures of Anhui by the end of 1997 and the measles surveillance system was established at the beginning of 1998. IgM antibody of measles and rubella was tested with ELISA and IgG antibody was examined with HI test. RESULTS: The positive rate of measles antibody among children was 100%, GMT increased from 1:18.97 to 1:43.45 after the campaigns. Measles cases in the three districts from January to June 1998 had a 87.71% decrease, comparing with the same period of 1997 with measles outbreak avoided. There were 105(58.01%) cases of measles with positive IgM and 32(17.68%) cases of rubella with positive IgM out of 181 testees having fever and rash illnesses. CONCLUSION: Results indicated the programs as catching-up immunization campaigns on measles and keeping high level of MV immunization, establishing measles surveillance system with improvement of its sensibility together with rubella control program all seemed to be important strategies in speeding up the positive outcome of measles control. PMID- 10682512 TI - [Evaluation on the establishment and operation on neonatal tetanus surveillance program in Hainan Province]. AB - OBJECTIVE: In order to learn the actual morbidity and influencing factors of neonatal tetanus(NNT) and to provide scientific basis for NNT elimination. METHODS: A surveillance program was set up in 1997 in Hainan Province and to operate at the same time. RESULTS: 216 NNT cases were investigated in 1997. The number was higher than reported 78 cases. The result from surveillance showed that parturition at home and the old delivering method were the main influencing factors 88.89% of mothers of NNT cases had no TT immunization history. The surveillance data of Sanya City and Qiongzhong County indicated the NNT morbidity in 1997 had declined 61.29%, 82.05% than that in 1996 respectively due to the improvement of TT immunization rate. CONCLUSION: The study result proved that the most economical and effective measure to eliminate NNT is to increase TT immunization rate of women at child-bearing-age. It was practical integrate the NNT surveillance program into AFP surveillance system and operate along with it. PMID- 10682513 TI - [A large-scale study on the safety and epidemiological efficacy of Japanese encephalitis (JE) live vaccine (SA14-14-2) in the JE endemic areas]. AB - OBJECTIVE: To measure the safety and epidemiological efficacy of Japanese encephalitis live vaccine(SA14-14-2). METHODS: Guoyang and Mengcheng counties in Anhui Province, were chosen as observed spots where high incidence of JE was noticed in China. 1-6 years old children in two counties were inoculated with vaccine manufactured by Chengdu Biological Products Institute in the beginning of 1992. All children of 1-6 years old received one primary dose of live vaccine and the children at one or two years old were respectively given one dose for primary or booster vaccination in 1993-1996. Side effects of live vaccine had been under surveillance for five years for its safety JE cases during the epidemic season (Jun to October), were diagnosed clinically and serologically. RESULTS: Data on the safety and efficacy of the JE live vaccine(SA14-14-2) are listed as follows:1) During 1992-1996, a total number, of 335,941 children at 1-6 year old were vaccinated. No vaccine-associated encephalitis, meningitis or other serious adverse events were observed. 2) The incidence of JE case has greatly declined since the beginning of large-scale vaccination. The average JE morbidity dcreased from 11.34/100,000 in 1987-1991 to 2.74/100,000 in 1992-1996(P < 0.005). 3) The incidence of JE case in 1-6 years old in that period reduced as well. The average JE morbidity decreased from 56.24/100,000 to 13.83/100,000 in Guoyang and from 44.57/100,000 to 16.94/100,000 in Mengcheng counties respectively(P < 0.005). Most of the JE cases (94%) occurred in the unvaccinated children including all the 18 death cases. 4) Serum antibody response to immunization were measured by plaque reduction neutralization test. Neutralizing antibody seroconversion after one single vaccination were 83.87%-94.74%. CONCLUSION: The results further confirmed that JE live vaccine is safe for children and effective for prevention from JE disease in JE endemic areas. PMID- 10682514 TI - [The investigation of natural antibody level to Haemophilus influenzae type b polysaccharide in China]. AB - OBJECTIVE: Investigation of natural antibody level to Haemophilus influenzae type b polysaccharide in China. METHODS: 1,596 sera samples from cord (55 sera), infants (1,383 sera), children (66 sera), adult (52 sera) and elderly (40 sera) were collected from different provinces and the natural antibody level to PRP was measured with a standardized radio immunoassay 125I-labeled PRP. RESULTS: Antibody level of infants under 5 year olds in different provinces was different. Levels of infants from Gansu, Guizhou, Jiangxi province were lower (< or = 0.07 microgram/ml) than from Shanhai, Guangxi and Heilognjiang. Comparison of antibody levels with different age groups showed that infants aged 6 months to 5 years having the lowest antibody level (< 0.15 microgram/ml). The level increased rapidly during 6-10 years of age (> or = 1.0 microgram/ml). The levels of adults and elderly were about 1-1.5 micrograms/ml. CONCLUSION: General immunity against Hib disease were low in Chinese population. Children aged 6 months to 5 years old with lowest natural antibody level to PRP were the population under high risk. PMID- 10682515 TI - [Analysis on characteristics and risk factors of acute myocardial infarction in younger adults]. AB - OBJECTIVE: To investigate the risk factors and characteristics related to acute myocardial infarction (AMI) in younger adults. METHODS: Clinical data were analyzed in 55 patients under 40 years old with AMI and compared with 1,097 controls of older age group (> or = 40 years). RESULTS: Results showed that the family history of coronary heart disease, smoking and alcohol intake were more frequently seen in younger age group than those in control group with odds ratios OR = 1.4, OR = 1.6 and OR = 1.6, respectively. Younger age group had a higher rate of noticeable inducing cause and the major causes were exhaustion and physically over-hurdened. A higher percentage of typical chest pain was found in younger patients. CONCLUSION: Smoking, alcohol drinking and genetic factor made up major risk factors of coronary heart disease in younger adults in this study, however reducing the amount of smoking and alcohol drinking, prevention of fatigue and over excitement might have some preventive impact on the prevalence of AMI in younger population. PMID- 10682516 TI - [Cellular immunity and epidemiologic analysis of pediatric patients with Mycoplasma pneumonia]. AB - OBJECTIVE: To understand the of immuno-reactions of pediatrics patients with Mycoplasma pneumonia. METHODS: 90 patients suffered from M. pneumonia were administered and divided into three groups: mild group, severe group, and normal control group. T cell subset parameters, natural killer cell and the serumsoluble interleukin-2 recepter of all of above were determined. RESULTS: Data showed:CD4 decreased at both acute and recovery stage of M. pneumonia, while CD8 remarkably increased (t = 2.63, 66, 2.77, 3.36, P < 0.05). SIL-2R level of all patients also greatly increased (t = 5.26, 3.19, P < 0.01), especially in the serious group. CONCLUSION: There are disturbances of cell-immune in M. pneumonia. The level of SIL-2R can serve as monitor on the degree of severeness of M. pneumonia. The incidence M. pneumonia appeared highest in the 10-14 year-old group. PMID- 10682517 TI - [Measurement of serum tumor necrosis factor-alpha and interleukin-8 in children with low respiratory tract infection (LRTI) caused by Mycoplasma pneumonia]. AB - OBJECTIVE: To investigate the immunopathological mechanism of LRTI caused by Mycoplasma pneumonia. METHODS: Serum tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) from 30 patients with LRTI caused by Mycoplasma pneumonia, were measured with sequence saturated solution competitive radioimmunoassay and sandwich ELISA. RESULTS: The level of serum TNF-alpha and IL-8 of patients suffering from LRTI caused by Mycoplasma pneumonia were magnificantly higher than that of healthy children. Meanwhile, a positive relationship between serum TNF alpha and IL-8 was noticed. CONCLUSION: TNF-alpha plays an important role in LRTI caused by Mycoplasma pneumonia, and serves the key factor to the elevation of IL 8. PMID- 10682518 TI - [Argument over meta-analysis in medicine]. PMID- 10682519 TI - [Epidemiologic study and prevention of AIDS in China]. PMID- 10682520 TI - [Progress on the study of prevention and control of anthrax in China]. PMID- 10682521 TI - [Marked success of prevention and control of sex transmitted diseases in pastoral area of Inner Mongolia Autonomous Region in China: epidemiologic survey and serologic follow-up study of syphilis]. PMID- 10682522 TI - [Study of superinfection of HBV and HCV]. AB - OBJECTIVES: To understand the situation in hepatitis B patients coinciding with HCV and to explore its influence on HCV on the replication of HBV. METHODS: Using ELISA, 712 hepatitis B patients were tested for serum anti-HCV and markers of HBV. RESULTS: Of the 712 patients, anti-HCV positive rate was 14.47% with the highest 48.98% in patients with severe hepatitis and the lowest 3.25% in patients with acute hepatitis. Markedly different anti-HCV positive rates (P < 0.001) in patients of different clinical stages were discovered. The more severe the case with longer the course, the higher the anti-HCV positive rates. In patients with superinfection of HBV and HCV, serum HBsAg, HBeAg and anti-HBcIgM positive rates were lower than those in patients with hepatitis B (P < 0.001, P < 0.001 and P < 0.05) but the anti-HBe positive rates were higher. All the differences showed an obvious statistical significance. CONCLUSION: Hepatitis B coinciding with HCV infection is responsible for the deterioration of the disease and towards its formation of its chronic phase as well as for the inhibition of HBV replication. PMID- 10682523 TI - [Analysis on the mis-reported cases of infectious diseases in Hainan Province]. AB - OBJECTIVE: To investigate the reasons for mis-reporting cases and to furnish theoretical foundation for the control of infectious diseases. METHOD: 15 hospitals and 2,937 residents were selected and studied through random and systematic sampling methods in Hainan Province. RESULTS: Data showed that the mis reporting rate in clinics was 44.40%, among which the private-ran clinics had a highest mis-reported records. They not only failed to fill in the forms of reporting infectious diseases, but also neglected to report epidemic information. Malaria, hepatitis, syphilis and gonorrhea were among the highest mis-reported diseases with malaria the most (91.73% mis-reporting rate). Remarkable difference was noticed in various hospitals (P < 0.01). CONCLUSION: The author concluded that many infectious diseases were seriously mis-reported, indicating the reporting system should be improved and the quality be enhanced. PMID- 10682524 TI - [Epidemiological analysis of the fires happened in China from 1950 to 1994]. AB - OBJECTIVE: Approaching the harmful factors and the preventive steps of fires for controlling and preventing fires from harming people. METHOD: Descriptive analysis was used to review 2,905,504 fires-cases in China from 1950 to 1994. RESULTS: On average, there were 3,432 people died and 6,726 injured in 64,567 fires per year. From the middle of 70's, the means of injured and death rates decreased slightly, but have gone up a little after 80's. The economical loss has risen greatly year after year, reaching the maximum of 1.3 billion Yuan (RMB) in 1994. The loss of a fire case in 1994 was more than twice than that in 1990. 89.4% fires were caused by man's faults. The fire cases in Sichuan, Yunnan, Jiangshu, Zhejiang and Guangdong Provinces were more serious than the average, of which Guangdong was the worst. Data from 17 departments showed that, the fire frequency in residential quarters of cities and countryside were highest (24%), and the number of injury and death accounted for 28.4%. The fire happens frequently in winter, and least in summer. CONCLUSIONS: The problem caused by accounted fire has not been controlled effectively. PMID- 10682525 TI - [A study of genetic epidemiology on child bronchial asthma]. AB - OBJECTIVE: To examine the relationship between child bronchial asthma and genetic fastor. METHODS: A case-control study including 372 pedigree was carried out. RESULTS: Child bronchial asthma had obvious familiar clustering (chi 2 = 24.8, P < 0.01). The prevalence rate of the first and the second degree relatives of proband was significally higher than that of the controls. The segregation ration of asthma was 0.14 (95% CL 0.098-0.18). The h2 of the first and second degree relatives of asthma were 78.18% and 55.02%, of which 83.64% for female and 73.48% for male. CONCLUSION: The genetic model of asthma belonged to polygenetics. Genetic fastor was a main risk fastor for asthma, especially for female patients. PMID- 10682526 TI - [Prevalence study of congenital heart disease in children aged 0-2 in Zhejiang Province]. AB - OBJECTIVE: To find out the prevalence rate of Congenital Heart Disease (CHD) among infants and toddlers. METHODS: Heart auscultation and echocardiography examination on children aged 0-2 were examined in 13 cities (or counties) in Zhejing province. RESULTS: Findings showed that the prevalence rate in children aged 0-2 was 3.72/1000. The prevalence rates of CHD were quite different among age groups with the highest (5.54/1000) in age group 0, followed by 3.36/1000 in age group 1 and lowest (2.66/1000) in age group 2. No significant difference of prevalence rates was found between different sex. Ventricular septal defects (59.4%) was noticed as the most common lesion. CONCLUSION: The evidence indicated that CHD is one of the most important problems of public health in China. Preventing its occurrence by conducting CHD surveillance and its etiologic research will have great significance for enhancing the qualities of life of children. PMID- 10682527 TI - [Investigation on the prevalence and influencing factors to anemia in women at reproductive age]. AB - OBJECTIVE: To investigate the prevalence of anemia in women of reproductive age and to analyze the influencing factors in anemia. METHODS: A total number of 1,529 women aged 15-49 years old including workers, farmers, cadres and students were tested with Hb and FEP and investigated through questionnaire including related influencing factors. Statistical analysis was performed using SPSS/PC statistical software. RESULTS: Mean value of Hb was 116.35 g/L (SD 14.67 g/L) in 1,529 cases and the prevalence rate of anemia was 31.2%. Majority of the anemia identified belonged to the 'iron deficiency type'. Influencing factors on anemia included occupation, education, marriage status, menstruation, status of family expenses and physical exercise. CONCLUSION: The prevalence rate of anemia in reproductive aged women was high, thus more attention should be paid. In order to lower the incidence of anemia, preventive and intervenient measurements should be conducted accordingly. PMID- 10682528 TI - [Markov process method in analyzing longitudinal quality of life data]. AB - OBJECTIVE: To present a method to analyze longitudinal quality of life data and discuss some relevant questions. METHOD: Based on the theories of Markov process, the Markov method was presented and used to analyze the change of quality of life status for 212 cases being detoxificated drug addicts and their influence factors. RESULTS: The intensity coefficients of status transient for 1-->2 and 2- >3 for occupation were 0.7912 and 0.6797 respectively; the coefficients for status 1-->2 for the detoxification time was 0.5663 and that for 2-->3 for the detoxification method was 0.2765. CONCLUSIONS: The Markov method is good to analyze longitudinal quality of life data for it can analyze both the transient probabilities between any two status of quality of life and their influence factors. The results showed that the occupation of drug addicts, the detoxification time and method can affect the change of quality of life. PMID- 10682529 TI - [Study on the association and significance between trace elements and rectal cancer]. PMID- 10682530 TI - [Analysis on epidemiological feature of injuries in civic students of Jiangmen]. AB - OBJECTIVE: In order to set up intervention measures in preventing injuries. METHODS: An analysis on the epidemiological feature of injuries in 9 civic middle and primary schools with 3,988 students involved was carried out in July 1998. RESULTS: The results showed that the total rate of 12 kinds of injuries was 50.55%. Among which, 5 kinds of injuries took the first 5 places: injuries from falls (32.15%), knife-cutting or sharp weapon hurt (21.99%) bumps (17.05%) traffic accidents (12.51%) burns and scalds (11.43%). CONCLUSION: The rate of injury was related closely with age parent's culture background and the number of children in families, but was not significantly related to sex, parent's occupation and the condition of living with relatives. Strengthening supervision, safety health education and increase awareness on self-protection were the most important points. Some intervention measures were being put into practice. PMID- 10682531 TI - [Long-term efficacy study of hepatitis B vaccination in newborns--results of 11 years' follow-up]. AB - OBJECTIVE: To evaluate the long-term efficacy of hepatitis B(HB) vaccination in newborns and the need for a booster dose. This research is one of the longest HB vaccine follow-up studies in the world with its subjects came from a program of universal infant HB vaccination. METHODS: Children who were born in 1986 and immunized with hepatitis B vaccine at birth were followed up at least once a year. Serum HBsAg, anti-HBc and anti-HBs were tested. At the 5th year after the first dose the prevalence of hepatitis B infection in the children in other district who were also born in 1986 and remained unvaccinated was surveyed as external controls. Random sampling was applied and the possible bias was analyzed. The trends of the positive rates of serum HBsAg, anti-HBs and anti-HBc in the immunized cohort were studied. With external control, the long-term efficacy of HB vaccination was calculated. RESULTS: The positive rates of HBsAg in the vaccine group from the first to eleventh year were 0.46%-0.98% and were below than those of baseline and external control. HBsAg rates in the cohort at different ages were similar with an average of 0.70%(25/3 578). The long-term efficacy of newborn vaccination was 85.42% (95% confidant interval: 70.11%-100%). CONCLUSIONS: The efficacy of HB vaccine was long-lasting and a booster dose was not necessary at least up to age 11 years. PMID- 10682532 TI - [Gene therapy of AIDS]. PMID- 10682533 TI - [Epidemiologic study of risk factors of gallstone]. PMID- 10682534 TI - [Global eradication of small-pox: historical fact, experiences and enlightenment]. PMID- 10682535 TI - [Progress in the study of spotted fever in China]. PMID- 10682536 TI - [Molecular-epidemiological analysis of HIV-1 initial prevalence in Guangxi, China]. AB - OBJECTIVE: In April 1996, HIV-1 infection was first found among both commercial blood donors and IDUs in Guangxi. In order to identify the source of HIV-1 transmission and analyze the trend of HIV-1 epidemic, the study was carried out. METHODS: HIV-1 genetic subtypes were determined by peptide enzyme immunoassays (PEIA), RT-PCR and DNA sequencing. RESULTS: Four subtypes of Group M HIV-1 were found in Guangxi:subtype B' (Thai B), C,D and E. Subtype E and C (1 person) were circulating among IDUs and hetrosexual, while subtype B' and D (1 person) were among commercial blood donors. Subtype B' infections were discovered among a group of commercial blood donors and one of them was infected by subtype D. CONCLUSION: It is the first report of subtype D infection and subtype E that were identified in China. This observation indicated that subtype E HIV-1 was spread into China from Southeast Asia, and a new epidemic region with subtype E HIV-1 would emerge in southern China. Serotyping might be a useful screening method for HIV-1 molecular epidemiological analysis. PMID- 10682537 TI - [Investigation on the effect and strategy of polio eradication in Hainan Province]. AB - OBJECTIVE: Hainan Province was a hyperendemic area of poliomyelitis, with an average incidence rate of 0.28 per hundred thousand before 1993. A series of strategy was developed for polio eradication in the whole province in July 1993. METHODS: Health education, regular and strenthening immunization programs, AFP surveillance. RESULTS: 1. OPV vaccination rate increased to 97.93%, 2. incidence rate of polio had a 80.93% reduction, 3. seasonal peak of epidemic was cut down, 4. endemic cycle was broken and 5. the endemic area became smaller. The number of counties (cities) where polio cases occured reduced for 81.33% in 1993 than that in 1989. Four boosters had been implemented from 1993 to 1997, to eliminated blank spots of immunization. The average geometric antibody of titre increased to 1:400 and above. After the establishment of polio surveillance system in 1991, 1. the reporting rate of non-AFP increased from 0 to 1.27 per hundred thousand, 2. rate of double specimens collection increased from 0 to 87.10%, 3. positive rate of virus isolation from 23.07% to 34.80%, 4. the positive specimens were appraised by National Polio Surveillance Center and no wild strain was found. Health education has made remarkable impact and immunization knowledge among parents had a 75 percentage point increase. CONCLUSION: Results showed that strategy of polio eradication has made positive effects and provided valuable experiences for the control or elimination of diseases which can be prevented by vaccines. PMID- 10682538 TI - [An effective measure to prevent and control STDs through epidemiological surveillance on sex-related criminals]. AB - OBJECTIVE: To study the prevalence and trend of sexually transmitted diseases (STDs) among sex-related criminals and to strengthen the strategy on prevention and control of STDs. METHODS: STDs surveillance on 956 criminals at selected sites in Beijing had been consecutively monitored during the period of 1996-1997. Data was then collected and analyzed. RESULTS: Out of the 956 criminals, 317 (33.2%) cases were identified as having STDs. Among them NGU took the first place and followed by gonerrhea and syphilis. CONCLUSION: The results revealed that STDs had become one of the major infectious diseases in Beijing and its incidence increased annually. Epidemiological surveillance on sex-related criminals is one of the effective measure in reducing the sources of infectious. PMID- 10682539 TI - [Study on hepatitis G virus infection]. AB - OBJECTIVE: To find out the situation of HGV infection in Shandong Province, and to explore the relations between HGV infection and HCV or HBV infection. METHODS: Enzyme linked immunosorbent assay (ELISA) was used to determine the serum anti HGV in 1,082 patients with viral hepatitis, 77 patients with non A-E hepatitis and 361 blood donors. RESULTS: 53 patients whose serum anti-HGVs were positive (positive rate: 3.49%) were noticed. The anti-HGV positive rate (8.93%) in patients with Hepatitis C was remarkably higher than that (3.32%) in patients with Hepatitis B (chi 2 = 8.80, P < 0.01). The anti-HGV positive rate (4.82%) in patients with chronic hepatitis was significantly higher than that (0.79%) in patients with acute hepatitis (chi 2 = 10.79, P < 0.01). The anti-HGV positive rate (8.00%) in patients with severe hepatitis was obviously higher than that in patients with acute hepatitis (chi 2 = 10.23, P < 0.01). CONCLUSION: The manifestations of HGV infection can be expressed as virus-carriers, subclinical infection or various clinical types. Patients with Hepatitis C were more subjective to be overlapped with HGV than the patients with Hepatitis B; moreover, HCV or HBV infection superinfected with HGV is associated with exacerbation of patients' condition and the formation of chronic infection. PMID- 10682540 TI - [Study on seroprevalence of Helicobacter pylori infection among upper digestive tract cancer patients and their kindreds]. AB - OBJECTIVE: In order to analyse the association between Hp infection and the risk of upper-digestive tract cancer. METHODS: In Huaian and Pizhou cities, Jiangsu province, Hp IgG quantitative-enzyme-immunoassay methods was used to identify IgG to H.pylori in the serum of 312 cases of upper-digestive tract cancer patients and their kindreds. RESULTS: (1) The level of IgG to H. pylori and the IgG positive rate (50.0%) in gastric cancer patients were both higher than that of cardia and esophageal cancer patients (P > 0.05, chi 2 test) but no significant differences were observed between cardia and esophageal cancer patients, 3 types of cancer patients and their kindreds; (2) the overall positive rates of both patients and kindreds in gastric cancer families (27.1%) were significantly higher than that of cardia or esophageal cancer families (P < 0.05) but no significant differences were observed between cardia and esophageal groups. CONCLUSION: H.pylori infection was not thought to be correlated with the development of gastric cancer, although higher clustering of Hp infection in families afflicted with gastric cancer was noticed. PMID- 10682541 TI - [Matched case-control study for detecting risk factors of breast cancer in women living in Chengdu]. AB - OBJECTIVE: To detect the risk factors of breast cancer in women living in Chengdu in order to identify high risk population and to conduct proper interventions. METHODS: A matched case-control study was performed in 265 cases with breast cancer who lived in this area. RESULTS: In the univariate analysis, menarche to menopause period > or = 35 years, taking oral contraceptives, history of benign breast diseases, passive smoking, and syrup drinking are the statistically significant risk factors of breast cancer, whereas breast feeding and soybean food consumption are protective factors. Conditional logistic multivariate analysis revealed that history of benign breast diseases and passive smoking are risk factors while soybean food products and breast feeding are protective factors. CONCLUSION: The risk factors of breast cancer in women living in Chengdu are history of benign breast diseases and passive smoking. PMID- 10682542 TI - [A case-control study on the dietary risk factors of upper digestive tract cancer]. AB - OBJECTIVE: To understand the effect of dietary factors in Yangzhong, Jiangsu Province-a high prevalence area in China. METHODS: A case-control study on 209 cases of upper digestive tract cancer was conducted. There were 68 cases of esophageal cancer, 69 cases of cardiac cancer and 72 cases of other gastric cancers including 129 males and 80 females aged 35-79 under the study. RESULTS: It is revealed that intake of pickled vegetables increases the ORs of esophageal, cardiac and other gastric cancers (OR = 2.82, OR = 5.17, OR = 2.92, respectively). It is also concluded that the intake of leftovers can elevate the ORs of esophageal and cardiac gastric cancer (OR = 1.88 and OR = 1.90) and over consumption of salt also elevates the OR of cardiac cancer (OR = 1.87). However, drinking green tea may decrease the ORs of esophageal and other gastric cancers (OR = 0.20 and OR = 0.28) while fruits consumption may reduce the OR of esophageal cancers (OR = 0.51). CONCLUSION: Tumors from upper digestive tract have some relations with diet factors but the effects vary with the differences of tumor sites, dose of exposure and area, etc. PMID- 10682543 TI - [An investigation on the prevalence of diabetes mellitus among 336 health workers]. AB - OBJECTIVE: To find out prevalence of diabetes millitus among health workers. METHODS: Physical examination was conducted among 336 health workers. Blood glucose was determined by routine glucokinase method. RESULTS: Results showed that IGT was 26.49%. According to the WHO diagnostic standard, the prevalence rate of diabetes was 9.23% (31/336) with type II 96.77% and type I 3.22%. Among diabetes cases, 61.29% (19/31) was newly diagnosed. The prevalence rates of IGT and diabetes were related to age and both BMI (Trend chi 2 test: P < 0.01) and family history were important factors (OR = 10) 42.11% of the new cases had complications. CONCLUSION: It is important to provide health workers with more knowledge on the prevention of diabetes. PMID- 10682544 TI - [A case-control study on the risk factors in naevus fusco-caeruleus zygomaticus]. AB - OBJECTIVE: To determine the risk factors of naevus fusco-caeruleus zygomaticus, a case-control study was conducted in 100 cases. METHODS: The selection of major risk factors was through logistic regression and chi 2-test. RESULTS: It was suggested that the high exposed rate in the family history of objectives was related to "genetic susceptibility". Histories of using cosmetics and exposure to ultraviolet ray were identified as environmental risk factors. However, there was no single factor that could completely explain the etiology of naevus fusco caeruleus zygomaticus. CONCLUSION: We believe that coordinating effects of both susceptibility in genetics and environmental risk factors may play important roles in the pathogenesis of naevus fusco-caeruleus zygomaticus. Thus combined research on the location of gene mutation with environmental factors is important in the discovery of the etiology of naevus fusco-caeruleus zygomaticus. PMID- 10682545 TI - [The epidemiological study on limb deformities among children in Guangdong Province]. AB - OBJECTIVE: To study the incidence of limb deformities of children in Guangdong province. METHODS: Physical examination on lined-up students and looked over one by one. Detailed examinations of orthopaedics or related subjects were performed on those with susceptive deformity. RESULTS: Results showed that there were 655 cases of limb deformities (0.64%) in 102,313 pupils of 7 to 14 years old, in which 550 cases (83.97%) needed surgical intervention. The incidence of deformity in economically poor areas was 2.88 times higher than those in advanced areas. CONCLUSION: Prevention and cure of limb deformities of children, especially those in rural area, should be addressed and emphasized. PMID- 10682546 TI - [A serological investigation of Legionella infection in six-species of poultries and domestic animals in Luzhou City, Sichuan Province]. AB - OBJECTIVE: To understand the prevalence rates of Legionella infection in poutries and domestic animals in Luzhou city. METHODS: Serological investigation of antibodies against Legionella pneumophila serogroups 1-14 and Legionella micdadei was carried out, using microagglutination test (MAT) in six species of poutries and domestic animals. RESULTS: Results showed that the infection with multiple serogroups was present. However, each species had its own serogroup with positive rates of different serogroups ranging from 2.00% to 28.75%. Positive rates of domestic animals and poultry accounted for 4.49% and 6.47% respectively. CONCLUSION: It was suggested that poultries and domestic animals were widely infected with Legionella. It is important to carry out epidemiological surveillance in these domestic animals, poultries and environment for a better control program of this disease. PMID- 10682547 TI - [Evaluation on the quality of life among cardiac carcinoma cases--fifteen years after surgical treatment]. AB - OBJECTIVE: To investigate factors affecting the quality of life among long-term survivors after resection of cardiac carcinoma. METHODS: Follow-up study on the quality of life among 68 cases with carcinoma of the cardia who survived over 15 years postoperatively in our hospital, was conducted between January 1996 and June 1998. RESULTS: Results showed that 88.2% of the patients had a high sense of well-being, 86.8% of them were able to eat ordinary food, 64.7% of them could carry on regular work as well as physical labor. 89.7% of the patients were satisfied with their quality of life and 77.9% of them felt excellent. CONCLUSION: The postoperative mental status and the occurrence of long-term complications in digestive tract played an important roles in affecting the quality of life. PMID- 10682548 TI - [The use of risk factors scoring method in screening for undiagnosed diabetes in general population]. AB - OBJECTIVE: Based on an epidemiological survey of NIDDM in the general population, two sub-population groups were selected to develop a screening method on risk factors to identify people at increased risk for undiagnosed diabetes. METHODS: Logistic regression analysis on the original data of population A was carried out to screen the main risk factors of diabetes. The score of the variable was determined based on the OR value and the aggregate score was used to predict the risk of undiagnosed diabetes. Both validity and effectiveness of the method were evaluated in population B. RESULTS: Results showed that the risk of having diabetes in the population increased along with aggregate scores of the method. Trend chi 2 test showed statistical significance (P < 0.01). When the threshold value was set up at 7, both sensitivity and specificity for identifying undiagnosed diabetes were 74.3% and 63.2%, respectively making the positive predictive value 4.2% and negative predictive value 99.2%. CONCLUSION: The benefits of this screen method seemed to be simple, economical and helpful to obtain a satisfactory response rate. The method could be used for health education and to identify people (community) at high risk for potential diabetes. It was predicted that this screening method could serve as an effective and useful tool for mass screening of NIDDM. PMID- 10682549 TI - [Study on double antigens sandwich enzyme immunoassay for detection of Brucella specific antibodies in human and animals]. AB - OBJECTIVE: In order to develop practicable methods for serodiagnosis, surveillance and epidemiological surveys in human and animals Brucellosis. METHODS: A double antigens sandwich Enzyme Immunoassay (DAgS-EIA include DAgS ELISA and DAgS-DIEA) have been developed under the basis of first production conjugate of Brucella antigen with horseradish peroxidase. Serum samples from diagnosed patients, suspected patients of Brucellosis and sheep infected with Brucella in Henan and Hebei provinces as well as from experimental animals of infected with Brucella in laboratory detected through DAgS-EIA, I-ELISA, RBPT and SAT tests. RESULTS: Of the diagnosed and suspected Brucellosis patients, positive rates of DAgS-ELISA, RBPT, I-ELISA, DAgS-DIEA and SAT were 60.0%-62.9%, 48.6% 58.1%, 55.6%, 53.7% and 44.2% respectively; as for sheep with Brucella-infection, positive rate of the above mentioned 5 tests were 81.8%. CONCLUSION: Results showed that DAgS-EIA were not only specific and practicable but may use for Brucella specific antibodies detection in humans and various animals with only one conjugate of Brucella antigen with horseradish peroxidase. PMID- 10682550 TI - [Schistosoma japonicum ferritin: cloning, nucleotide sequencing, expression, and purification]. AB - Our previous work showed that immunization of mice with Schistosoma japonicum (Sj) immature eggs induced significant immunity against fecundity and embryonation of the parasite. The Sj adult cDNA library was screened by sera from rabbits against Sj immature egg antigen (RASjIEA). The genes encoding molecules which may induce immunity against fecundity/embryonation were chosen for further cloning and expression. First of all, RASjIEA was absorbed with E. coli lysate to remove cross reactive antibodies. The cDNA library was then immunoscreened using the routine method. The resulted positive plaques were rescreened till individual clones were confirmed. Phagemids were obtained using in vivo excision. The positive clones were amplified using PCR. The sizes of the genes were determined by agarose gel electrophoresis. After DNA sequencing of the genes cloned, Gene bank was searched and six different genes were identified from a total of 102 positive clones. One of six identified genes, Sj ferritin (SjFer) was chosen to subclone into pGMC vector. According to DNA sequences of Sj Fer and MCS (multiple cloning site) of the vector, forward primer (Fer/GMC1) and reverse primer (Fer/GMC2) were designed and used to amplify Sj Fer by PCR. The Sj Fer cDNA and expression vector pGMC were digested with BamHI and XhoI. The digested cDNA and pGMC were ligased by T4 DNA ligase to construct a recombinant which was then used to transform E. coli strain ER2566. The fusion protein GMCSF-Sj Ferritin was expressed in insoluble form, the inclusion body. Pellets were harvested and resolved in Tris-HCl buffer containing 8M urea. GMCSF-Sj Ferritin was purified by affinity chromatography using Ni-NTA resin. The molecular weight was determined by SDS-PAGE. This study first reports the gene encoding S. japonicum ferritin as a new candidate for schistosome vaccine. PMID- 10682551 TI - [Alterations of amino acids in rat cerebral cortex of infectious brain injuries]. AB - Concentrations of amino acids such as glutamate(Glu), glutamine(Gln), gamma aminobutyric (GABA), glycerin(Gly) in rat cerebral cortex were measured by o phthaldialdehyde method to clarify alterations of these amino acids in infectious brain injuries(IBI). The results were that Gln and GABA in groups treated by bordetella pertussis suspension(BP) 4h were increased compared to those in the group treated by the normal saline(NS) or the operative control(OC) and a positive correlation with water content or Evans blue content, respectively. In the 24h BP group, Gln was still increased; GABA and Gly were decreased compared to those in the NS or OC group. The findings suggest that GLu, Gln, and Gly play an important role and GABA may be a marker of injury in pathogenetic mechanism of IBI induced by BP. PMID- 10682552 TI - [Influence of ischemic preconditioning of heart on ischemia/reperfusion injury of rabbit lung]. AB - The influence of ischemic preconditioning of heart on ischemia-reperfusion injury of 16 Japanese big-ear-rabbit lungs were studied. The results showed: 1. Experimental group had a significant reduction in wet/dry weight ratio; 2. It had a significant increase in PaO2. 3. It had a minimal intraalveolar neutrophil infiltration in comparison with the control group. These data suggestes that the ischemic preconditioning of heart can lighten the ischemia-reperfusion injury of rabbit lungs. PMID- 10682553 TI - [Experimental study of FZBZ decoction enhancing chemotherapy efficacy for mice bearing with tumors]. AB - To study the mechanism of the increasing effect of Fuzheng Baozhen decoction (FZBZD) on chemotherapy. The BALB/C mice with transplanted human lung adenocarcinoma (SPC-A-1), sarcoma (S180) were treated in different groups. Comparing the group treated with chemotherapy (CTX), and the group treated with CTX plus FZBZD, the inhibiting tumor rate was higher (P < 0.01), the cAMP/cGMP ratio was increased by adding cAMP level in cancer tissue (P < 0.05), the level of serum TNF-alpha and MDA was decreased (P < 0.01), the activity of total SOD was improved (P < 0.01), the G0/G1 phase cells enhanced and S phase cells decreased in tumor tissue (P < 0.05). It suggested that FZBZD increased the effect of chemotherapy in different ways. PMID- 10682554 TI - [Oxidative DNA damage of cartilage in osteoarthrosis]. AB - In order to study molecular mechanism of osteoarthrosis, we used molecular techniques to detect DNA oxidative damage of cartilage. There were 10 samples from clinical operation including 5 cases of replacement of hip joint, 4 degenerative cartilage of knee osteoarthrosis, and 1 free body of knee joint. DNA from all samples was run out on an agarose gel and appeared classic nucleosomal ladders. (8-hydroxyl-2'-deoxyguanosine)/ml (8-OHdG) in DNA samples was measured by high-performance liquid chromatography with electrochemical detection (HPLC EC). The results showed that the levels of 8-OHdG were increased to 0.11-0.26 ng.ml-1, average 0.177 ng.ml-1. Normal controls were lower than 0.05 ng.ml-1. The high 8-OHdG in DNA led to a misreading of affected templates and base-pair exchange, which could be a sensitive indicator about oxidative DNA damage, which was produced by free radicals or other DNA damage agents. The result suggests that oxidative DNA damage may be the molecular mechanism of osteoarthrosis. PMID- 10682555 TI - [Relationship between expression of tumor suppressor protein p21 and p53 and cell proliferation in the gastric carcinoma]. AB - To study the relationship between the expressions of tumor supressor protein p21 and p53 and malignant growth of gastric carcinoma, 88 paraffin embedded specimens of gastric carcinoma and gastric ulcer were examined with immunohistochemical method. We found that the expression rate of mutated protein p53 in the gastric carcinoma was about 40% and the expression rate of p21 protein in gastric mucous carcinoma, undifferentiated carcinoma, poorly differentiated carcinoma was obviously low. The expression of PCNA in gastric adenoid cancer was very high. The results suggest that the low expression of p21 proteins and mutation of p53 proteins in gastric cancer cells play a certain role in the morbidity of gastric carcinoma, but its involvement in the malignant growth of gastric carcinoma cells is not obvious. PMID- 10682556 TI - [The prevalence of Chlamydia trachomatis and Ureaplasma urealyticum cervical infection in infertility women and the observation of therapeutic efficacy]. AB - OBJECTIVE: To study the prevalence of Chlamydia trachomatis (CT) and Ureaplasma urealyticum (Uu) cervical infection in infertility women, and therapeutic effectiveness of tetracycline and qianglimycin. METHOD: CT in cervical swab specimen was detected by cell culture, polymerase chain reaction(PCR) and immunofluorescent assay(IFA), Uu in cervical swab specimen was detected by culture, in a group of 145 infertility women, before and after treatment, and 45 women at productive age who attended obstetric and gynecologic clinic. RESULT: The positive rate of CT by cell culture, PCR, IFA(shell) and IFA(direct smear) was 62.7%, 66.8%, 64.8% and 36.5%, respectively, which was obviously higher than that of the control (P < 0.01). The positive rate of Uu was 33.1%, that of CT and Uu co-infection was 18.6%. Both were obviously higher than those of the control (P < 0.01). CONCLUSION: This study indicates that the prevalence of CT and Uu cervical infection in infertility women is high. CT and Uu cervical infection is closely related to female infertility. The therapeutic effectiveness of tetracycline and qianglimycin is not ideal. PMID- 10682557 TI - [Influence of dietary habits and body weight on blood uric acid in the elderly]. AB - Influence of dietary habits, body weight on blood uric acid was studied in 416 elderly people. The result showed that level of blood uric acid in the people who had habits of drinking alcohol, tea and taking hot foods was higher than that who never had those habits (P < 0.05 or 0.01). It also showed that level of blood uric acid was significantly increased in the over-weight or obesity people (P < 0.05). The hyperuricemia incidence in the over-weight or obesity people is 27.4 per cent, and it is 2 times and 3.4 times of the people with ideal weight and weak-weight, respectively. It is suggested that the patients with gout or hyperuricemia give up drinking alcohol, tea and taking hot foods for their health. Reducing body weight is one of the effective measures to prevent and treat gout or hyperuricemia in the elderly. PMID- 10682558 TI - [Efficacy of epimedium compound pills in the treatment of the aged patients with kidney deficiency syndrome of ischemic cardio-cerebral vascular diseases]. AB - One hundred-twenty aged patients with kidney deficiency syndrome of ischemic cardio-cerebral vascular diseases were treated with Epimedium compound granules. The results showed that after therapeutic period the total and marked effective rates were 96.7%, 39.5% respectively in treatment group. The rates of improvement were 70% in electrocardiogram of the patients with coronary heart diseases, and 75% in electro-encephalogram of the patients with cerebral arterosclerosis. The therapeutic effectiveness in treatment group was better than that in control group treated with Su-Guan-Bian. According to experimental observation, the therapeutic effectiveness was related to the facts that Epimedium compound granules lower blood lipid have anti-free radicals and adjust balance between prostacyctin I2 and thromboxane A2(TXA2/PGI2). PMID- 10682559 TI - [A prospective study on etiologic bacteria in 200 patients with pneumonia]. AB - The etiologic agents in 200 patients with pneumonia were studied by the bacterial culture of sputums obtained from the protected single catheter brush or quantitative expectoration at one morning or three-morning expectoration. Two hundred patients were divided into 3 groups. Group 1 was Nosocomial pneumonia (NP patients). Group 2-1 and Group 2-2 were community acquired pneumonia (CAP patients). All cases in Group 1 and Group 2-2 suffered from significant underlying diseases while Group 2-1 did not. Gram-negative bacilli(GNB) were isolated from the specimens in Group 1 (87%) and Group 2-2 (75%), respectively. Pseudomonas (30.8%) and klebsiella (20.5%) were the predominant bacteria (in Group 1 and pseudomonas bacteria) in Group 1 and pseudomonas (27.3%), acinetobacter (23%) and kledsiella (18%) were the major etiologic agents in Group 2-2. The commonest pathogens in Group 2-1 were gram-positive cocci (75%), in which streptococcus (38%) and staphylococcus aureus (25%) were the dominant agents. Compared with Group 2, Group 1 suffered from more mixed bacteria and the agents presented severer drug-resistant. The prognosis was worse in Group 2-2 than in Group 2-1. The results showed that the GNB pneumonia was more common in the cases who had underlying disease, no matter whether the pneumonia was NP or CAP. These patients had more trouble on their antibiotic therapy. Thus it is important that doctors should use vigorous antibiotics timely while treating these patients' underying diseases. PMID- 10682560 TI - [Relationship between ventricular arrhythmia and blood lipid in hypertensive patients with left ventricular hypertrophy]. AB - The possible relationship among ventricular arrhythmia(VA), blood lipid(BL) and left ventricular hypertrophy(LVH) were investigated in 92 patients with essential hypertension(EH). The results showed that the incidence of VA and complex VA and ventricular tachycardia(VT) were significantly higher in EH with LVH than those in EH with non-LVH. The incidence of VA and complex VA and VT were highest in hypertensive patients with LVH accompanied with hyperlipemia(HL), and VA seemed closely related to left heart structure and BL. The results suggest that LVH and HL are the two additive risk factors of occurrence of VA in patients with EH. PMID- 10682562 TI - [Clinical study on the orthodontic treatment of deep overbite with bite plane]. AB - Fifteen cases of patients with incisal Angel's Class II obserbite were orthodontically treated with bite plane and analysed with cephalometric radiograph. The results showed that bite plane can correct deep overbite rapidly (within 6 months) and effectively. The mechanism of bite plane was to raise the posterior teeth (1.1 mm) and to lower the anterior teeth (0.8 mm). PMID- 10682561 TI - [Relationship between sex hormone levels and blood lipids/immunity in perimenopausal women]. AB - To investigate the relationship between sex hormone levels and blood lipids/immunity and to evaluate the therapeutical effects of nylestriol, 96 women without coronary heart disease(CHD) were studied during their perimenopausal period. The estimation of serum biochemical components included serum 17 beta estradiol(E2), testosterone(T), total cholesterol(TC), triglyceride (TC), high density lipoprotein cholesterol(HDL-C), low density protein cholesterol(LDL-C), apolipoprotein A-I(ApoAI), apolipoprotein B(ApoB), lipoprotein (a)[Lp(a)], immunoglobulin G(IgG), and IgG antibody against cardiolipin(ACAIgG). Thirty-six postmenopausal volunteers were divided into two groups and randomized to treat with either 2 mg nylestriol or placebo. In postmenopausal women, serum levels of E2, E2/T, HDL, and ApoAI decreased, while those of T, TC, TG, LDL, ApoB, Lp(a), IgG, and ACAIgG increased. Serum level of E2 was positively correlated to HDL-C and negatively correlated to TC, ApoB, LDL, IgG, and ACAIgG. Serum level of T was positively correlated to LDL, IgG, and ACAIgG and negatively correlated to HDL. In the nylestriol group, as compared with the results before treatment, serum levels of TG, TC, LDL, IgG, and ACAIgG decreased and that of HDL-C increased after treatment. We conclude that estradiol is a protective factor of CHD, whereas testerone is a dangerous factor. After menopause, the imbalance of the estradiol/testosterone ratio increases the incidence of CHD. Nylestriol is an effective substitute for estradiol to prevent CHD in post menopausal women. PMID- 10682563 TI - [Changes of plasma endothelin-1 in patients with congestive heart failure and the influence of metoprolol]. AB - To investigate the alterations of plasma endothelin-1(ET-1) in patients with congestive heart failure(CHF) and the effects of metoprolol on it, plasma ET-1 and norepinephrine(NE) were measured in 43 patients using radioimmunoassay and high-performance liquid chromatography methods. Twenty-four patients were treated with metoprolol plus the routine therapy while the others were received the routine therapy only. The findings were that levels of plasma ET-1 and NE increased before the treatment, and decreased after the treatment with metoprolol for 1 month. There was no alteration of plasma ET-1 or NE in the control group. PMID- 10682564 TI - [Chronic pulmonary heart diseases treated by fraxiparin]. AB - OBJECTIVE: The effects of fraxiparine with hypodermic on the treatment of patients with chronic pulmonary heart diseases(CPHD) at acute phase were studied. METHODS: Twenty patients were hypodermically injected with 0.4 ml fraxiparine per day for 7 days as a course. Fibrin fibrinogen degradation products (FDP and D dimer), antithrombin III (AT-III), PaO2 and PaCO2 were detected before and after the treatment. RESULTS: In the therapy group, FDP and D-dimer decreased after the treatment, AT-III increased. All indexes didn't alter in the control group. CONCLUSION: Fraxiparine is effective on patients with CPHD at acute phase. PMID- 10682565 TI - [Clinical significance of heart rate variability: analysis of silent myocardial ischemia]. AB - To study the potential role of dynamic electrocardiogram(DEG) in silent myocardial ischemia (SMI) patients, we measured the extension of ischemia and heart rate variation-time-domain analysis in 148 patients with SMI and 30 healthy controls by DEG and followed up all patients for 1.5 years. The results were that the extension of myocardial ischemia (the extension of ST segment depression, episodes of SMI attack, and the total ischemia time), the incidence rate of ventricular premature heat increased gradually and the difference of heart rate. Heart rate variant hinder(HRVI), and SDNN decreased gradually in SMI I, III, II types. And the extension of ischemia had positive correlation with the lowest heart rate and the incidence rate of ventricular premature beat, while negative correlation with HRVI and SDNN, indicating that the impairment of cardial autonomic nerves is associated with the extension of ischemia. Also, we found that 23 patients out of 148 patients with SMI died and the levels of HRVI and SDNN of the patients who died were lower than that of the survivals. We conclude that the heart rate variability can serve as a prognosis index of SMI. PMID- 10682566 TI - [Clinical study of fructose 1,6-diphosphate on myocardial ischemia/reperfusion injuries]. AB - Twenty ventricular septal defect patients who underwent cardiopulmonary bypass (CPB) were divided into 1,6-diphosphate group and controlled group (10 cases each). Blood samples were taken at pre- and post-cardiopulmonary bypass 30 min, 6 h, and 24 h to determine lactate dehydrogenase enzyme(LDH), creatine kinase enzyme(CK) and its isoenzyme(CK-MB) levels. We found that 1,6-diphosphate decreased the increasing levels of LDH, CK, and CK-MB after cardiopulmonary bypass. It is suggested that 1,6-diphosphate may attenuate myocardial ischemia/reperfusion injuries during cardiopulmonary bypass. PMID- 10682567 TI - [Influence of MP infection on immunologic function in patients with chronic obstructive pulmonary disease]. AB - The aim of this study was to detect Mycoplasma pneumoniae(MP) in sputum with chronic obstructive pulmonary disease(COPD) by using polymerase chain reaction and measured the level of sIL-2R in chronic pulmonary heart disease(CPHD) patients by Enzyme Linked Immnosorbert Assay(ELISA). At the same time, we also measured the level of the serum total IgE in CPHD patients. We determine the relationship between the sIL-2R and lymphocytes. The results showed that rate of MP infection in COPD patients was 34.59%. The rate of MP infection in CPHD patients and asthmatic patients were significantly higher than that in patients with chronic bronchitis. The level of sIL-2R in patients with CPHD who had MP infection increased significantly higher than that in CPHD patients which didn't have MP infection. There was a linear negative correlation between the level of sIL-2R and lymphocytes. The level of serum total IgE in CPHD patients with MP infection was higher than that in patients with other infection. These data suggest that MP is a common agent in COPD patients, particularly in CPHD and asthma. MP infection can impair the cellular immunity and immunotherapy combined with antibiotics may be more effective. PMID- 10682568 TI - [Relationship between IL-1 beta and TNF-alpha in subretinal fluids of rhegmatogenous retinal detachment with PVR]. AB - The concentrations of IL-1 beta and TNF-alpha in subretinal fluids from 49 patients with rhegmatogenous retinal detachment with proliferative vitreoretinopathy(PVR) were measured by Enzyme-linked immunosorbent assay (ELISA). The results showed that all subretinal fluids contained IL-1 beta, their concentrations were positive correlation with severity of PVR (r = 0.677) and no correlation with age, sex, holes, ranges and times of retinal detachment, TNF alpha was detected in 18/49 subretinal fluids including 2 patients with PVR-1 beta and 16 patients with PVR-C and the above cases. It suggested that IL-1 beta and TNF-a are involved in the pathogenesis of PVR. PMID- 10682569 TI - [Management of hepatic trauma in 267 cases]. AB - Two hundred and sixty-seven consecutive hepatic trauma in recent 17 years were analysed. They were classified by AAST classification: grade I and II 68 cases (25.4%), grade III 132 cases (49.44%), grade IV 52 cases (19.48%) and grade V 15 cases (5.6%) and 237 cases were operated in different procedures according to the severity of the trauma. More than ten modalities such as dedridement, suturing, repairment, abdominal drainage, billiary drainage, hepatic artery ligation, packing and pressing hemostasis, resection of traumatic parecchyma, vessel ligation with suturing in section etc were used, The morbidity and mortality were 20.6% and 8.9%, respectively. The other 24 cases were found no active bleeding during laporatomy and rehabitated with preservative management. The authors believe that effective management on time and intensive perioperative care are important to minimize the morbidity and mortality of hepatic trauma. Nonoperative management deserves enough attention. PMID- 10682571 TI - [Changes of serum myocardial enzymes in patients with malignant hematologic diseases]. AB - Myocardial enzymes including aspartate aminotransferase (AST), lactate dehydrogenase (LDH), HBD (LDH1 and LDH2), and creatine kinase (CK) and its isoenzyme were monitored in 106 cases of malignant hematologic diseases. The findings were that average values of LDH and HBD increased. There were 82.4% myocardial enzyme levels of 51 patients with leukemia, lymphoma, and myelodysplastic syndrome (MDS) returning to normal or making an obvious reduction after chemotherapy associated with drugs of heart toxicity, while there were increases of the myocardial enzyme levels before chemotherapy. Patients with the increasing of enzyme levels were only 3.9% after chemotherapy. After several courses of chemotherapy, the positive rates of the increasings of CK and CK-MB were higher than that of pretreatment. The results suggest that the injuries of myocardium are possible. PMID- 10682570 TI - [Influence of isoflurane and sevoflurane on metabolism of oxygen free radicals in cardiac valve replacement]. AB - To investigate effects of isoflurane and sevoflurane on the levels of lipid peroxides (LPO) in plasma and superoxide dismutase (SOD) in red blood cells during myocardial ischemia/reperfusion, and to study myocardial protection of these two volatile anesthetcs, 30 patients undergoing cardiac valve replacement were randomly divided into isoflurane (Group I), sevoflurane (Group S), and controlled group (Group C). Patients of Group I were anesthetized with isoflurane mainly, Group S with sevoflurane, and Group C with fentanyl. LPO and SOD levels were measured at five points before and during the operation. The results were as follows: SOD levels in all groups did not change significantly. LPO levels in Group C increased during the operation, while in other two groups didn't. Rates of automatical rebeating of hearts in Group C were lower than that of hearts in other two groups. It is suggested that isoflurane and sevoflurane can inhibit increasing of LPO levels to attenuate myocardial ischemia/reperfusion injuries mediated by oxygen free radicals during open-heart surgery. PMID- 10682572 TI - [Preliminary study of clinical scoring on prognosis of severe head-injury]. AB - In this article, 68 cases of severly head-injured patients scoring and prognosis evaluation were reviewed. We have come to a practicable severe head injury index (SHII) for first aid: the blood pressure (Bp), pulse rate (P), respiration rate (R), degree of consciousness and reaction of the pupils are five items taken for estimation of the patients. The highest total score was 25, the lowest was 5. If the score was lasted for 2-4 hours below 7, and first aid non-responsive in 4-6 hours, the mortality and morbidity rates will be high. When the score is more than 14, the first aid salvage will be meaningful. The first aid salvage will be meaningful. The first aid had significant measing especially for those patients whose score was over 18. The detailed method of clinical scoring is shown on chart 2. PMID- 10682573 TI - [Relationship between plasma NO and PaO2 of artery blood in the patients with chronic pulmonary heart disease]. AB - The plasma nitric oxide(NO) level in the 28 patients with chronic pulmonary heart disease and health controls were measured. The results showed that NO level in the exacerbating patients and stable patients was significantly lower than that in health controls. The NO level in the patients getting worse was markedly lower than that in the stable patients. The lower the oxygen pressure of the artery blood (PaO2), the lower the NO level. when the patient's situation was improving, the plasma NO level increased with rising PaO2. The plasma NO level in patients was associated with the PaO2. This study suggests that the sustained pulmonary hypertension caused by chronic hypoxia may relate to the reduction of NO synthesis and release, and that NO may play an important role in the pathogenesis in chronic pulmonary heart disease. PMID- 10682574 TI - [The applied value of BiPAP mechanical ventilation via facial of nasal mask before or after ordinary mechanical ventilation]. AB - To expore the applied value of BiPAP ventilator before or after regular ventilation, 44 patients who had indicators of regular mechanical ventilation and 4 patients who had difficulty of getting free from endotracheal intubation mechanical ventilation were ventilated with BiPAP ventilator via facial or nasal mask. The results showed that 13/44 patients had good responses and avoided receiving regular mechanical ventilation with endotracheal intubation or incision. BiPAP ventilation was also effective in patients who were dependent on regular mechanical ventilatin. PMID- 10682575 TI - [Bcl-2 mRNA expression in acute promyelocytic leukemia by RT-PCR]. AB - To check the anti-apoptosis gene bcl-2 mRNA expression in 20 APL patients by PCR, and analyse the relationship between bcl-2 mRNA and the PML-RARa isoform. The result is that the bcl-2 mRNA expression has no significance between the isoforms (P > 0.05). This shows that the PML-RARa transcription type has no influence on the clinical features and prognosis. PMID- 10682576 TI - [A new fluorescent scanning technique in quantitative determination of isoenzymes of creatine kinase]. AB - To increase the sensitivity and specificity and decrease the cost on determining the isoenzymes of creatine kinase (CK), including CK-MM, CK-MB, and CK-BB, we used the electrophoresis bath with different buffers to isolate the isoenzymes and used the fluorescent scanner to determine the content of each isoenzyme. We found that the concordance appeared in sensitivity and specificity in comparison with the MOPS electrophoretic system. There was a positive correlation between our method and MOPS, and the cost was much less than that of the MOPS. PMID- 10682578 TI - [Effect of different doses of succinylcholine chloride on its action duration during rapid anesthesia induction]. PMID- 10682577 TI - [Analysis of bacterium culture and drug sensitivity test in bile of biliary tract diseases]. PMID- 10682579 TI - [Therapeutic experience of primary inferior tracheal tumor. Report of 10 cases]. PMID- 10682580 TI - [Uses of valve patches for repairing the congenital heart defects with severe pulmonary hypertension]. PMID- 10682581 TI - [A clinical analysis of 40 patients with the unknown origin fever]. PMID- 10682583 TI - [A case of particular-long slow pathway conduction among triple A-V nodal pathway]. PMID- 10682582 TI - [A case of primary ovary non-Hodgkin's malignant lymphoma]. PMID- 10682584 TI - [A case of Tourette syndrome complicated with schizophrenia]. PMID- 10682585 TI - [Relationship between health status and work ability among elderly Al exposed workers]. AB - In this study, health status and work ability of 80 elderly male workers exposed to aluminum (Al) were assessed by physiological and neurobehaviral tests. The results showed that workers with cardiovascular or respiratory diseases had lower work ability index(WAI). Vital capacity and musculoskeletal function index(MSFI) were correlated closely with work ability. Mental performance capacity decreased with the increasing of age, and mental performance score was associated with work ability. It is suggested that vital capacity and MSFI may serve as index reflecting physical work ability of Al-exposed workers. Simple reaction time, visual memory, manual dexterity and pursuit aiming may reflect mental work ability. PMID- 10682586 TI - [Detection of sister chromatic exchange in workers exposed to coal tar pitch and to coke oven volatiles]. AB - In order to know the changes of genetic toxicological effects on workers occupationally exposed to polycyclic aromatic hydrocarbons (PAHs), sister chromatic exchange(SCE) was detected by the methods of peripheral lymphocyte culture in 23 workers exposed to coal tar pitch (CTP) and in 19 workers exposed to coke oven volatiles (COV) and 12 normal controls. The results suggested that the SCE in occupational workers was significantly higher than that in controls (11.31 vs 6.37, P < 0.001). The SCE in workers exposed to CTP and to COV was higher than that of control (10.27 and 12.58 vs 6.37) respectively. In workers exposed to CTP and COV, there were no differences of SCE for smokers and nonsmokers (P > 0.05). It is indicated that CTP and COV caused strong genetic toxicity and injury to chromosome. PMID- 10682587 TI - [Application and analysis of biochemical indices for the evaluation of antisilicosis treatment. Study on anti-silicosis therapy and its evaluation research group]. AB - The levels of serum Ceruloplasmin (Cp), Superoxide dismutase (SOD) and IgG of 296 silicosis patients treated by tetrandrine, polyvinylpridine-N-Oxide, hydroxypiperaquinoline phosphate and aluminium citrate were measured. Sera were collected before and after the 1st, 3rd and 6th therapy courses. 144 Silicosis patients without treatment were observed as controls. The levels of these three indices decreased by the end of treatment. The levels of SOD were fluctuated, which were increased after the 3rd course, but decreased after the 1st and 6th courses. The decrease of Cp, SOD and IgG consisted with the clinical effectiveness of the treatment, indicating that Cp, SOD and IgG were appropriate biochemical indicators for the evaluation of antisilicosis drugs. The quality control and the statistics standardization for data analysis are important. PMID- 10682588 TI - [Expression of p53 and ras p21 gene products in malignant transformed V79 cell induced by various organic components of DEPs]. AB - The effect of various fractions of diesel exhausted particles on the expression of mutant p53 antigen product and ras oncogene p21 product was examined by immunohistochemical method in malignant transformed V79 cell. The result showed that all fractions of diesel exhausted particles (DEPs) could significantly increase the expression of p53 products (P < 0.01). It demonstrated that the organic fractions of DEPs could induce the mutation of p53 suppressor gene in malignant transformed cells. But the expression levels of ras oncogene p21 product were not changed under the same experimental conditions (P > 0.05). PMID- 10682589 TI - [Ceruloplasmin gene expression in silicotic rat lung]. AB - In order to investigate the existence of extrahepatic ceruloplasmin (Cp) gene expression during silicotic process, molecular hybridization technique including dot blotting, in situ hybridization and Northern blotting was used. Twenty one days after silica dust intratracheal injection, Cp mRNA was detectable in lung, which was nearly double the content of that in the control (saline injection). In situ hybridization and Northern blotting revealed that the alveolar macrophages in silicosis could express Cp mRNA. It is concluded that lung is a prominent site for Cp gene expression during silicosis and Cp may play a previously unknown role in pulmonary injury or repair. PMID- 10682590 TI - [Antagonistic action of organic selenium on lead poisoning]. AB - Ninety white rats were treated with liquid lead acetate. When the concentration of lead in urine was more than (0.81 +/- 0.26) mg/L and blood lead was more than (4.8 +/- 0.33) mg/L, the rats were divided into three groups randomly. The oxygenic damage indicators in serum showed SOD activity decreased and MDA content increased. In one group of lead rats fed with an organic selenium compound (code FCQY) the SOD activities were high and the lipid peroxidation was inhibited compared with controls without selenium supplementation. The lead contents of bone, kidney and liver were measured. The results suggested that the organic selenium compound could interfere with the absorption and accumulation of lead. PMID- 10682591 TI - [Study on the psychological status of video display terminal operator]. AB - Occupational psychological test designed by the Labour Hygiene Department of West China University of Medical Sciences was applied to survey the mental health of 516 video display terminal (VDT) workers and 396 control workers in Chengdu. The results showed that mental health level of VDT workers was higher than that of controls. In the somatic disorder, anxious and hostile are increased obviously. With the prolonging of operating hours per week, somatic disorders, depression and obsession are increased, especially when VDT operating time is more than 30 hours per week and operating on VDT is more than ten years. This indicates that the operating hours should not be more than 30 hours per week. It is found by multiple factors analysis of data that the main adverse effects on mental health are age and VDT operating time. PMID- 10682592 TI - [Effect of manganese on the growth and development of rat offspring]. AB - Effects of Manganese exposure to pregnant rats on the growth and development of their offsprings were as follows: (1) the gain of body weight and brain weight in the high dose Mn-exposed offspring was significantly lower but the ratio of brain to body weight was significantly higher; (2) in Morris Water Maze Test, the average latency to find the hidden platform in the high dose Mn-exposed offsprings was obviously reduced during the first 5 days. But on the non-platform test day, a much longer length of swimming in the quadrant in which the hidden platform was located previously, although no difference was found for the total distance between them; (3) the immunoreactivity of glial fibrillary acid protein and the average density of its products in hippocampus of both the low and high dose Mn-exposed groups, especially of the high dose one, was significantly higher than that of the control group. PMID- 10682593 TI - [Relation between methylmercury chloride-induced programmed cell death and the development of nervous system in rats]. AB - The study was designed to reveal the role of programmed cell death (PCD, or apoptosis) induced by methylmercury chloride (MMC) in SD rat embryos, wish TdT mediated dUTP nick end labeling (TUNEL), Nile blue sulfate (NBS) vital stailing electron microscopy (SEM) and in vivo teratogenic methods (TEM). Pregnant rats were intraperitoneally injected at gestation day 9.5 with doses of 0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg/kg MMC and killed at 11.5 day later. Results showed that the PCD of embryonic nervous system was apparently dose dependent with MMC during rat head-fold stage. Observations under SEM and TEM showed that MMC caused pathologic changes of epithelia and organellae in embryonic brain such as atrophied or decreased microvilli cavernous damages and mitochondrial swelling and so on. Though MMC could result in developmental anomalies of embryonic brain and other organs, the main defect was open neural tube. The experiment study suggested that over PCD may be one of the teratogenic mechanisms of MMC on rat embryos, developing brain in particular. PMID- 10682594 TI - [Determination of tetradecyl pyridyl bromide in waste water by flame atomic absorption spectrometry]. AB - A indirect method for the determination of tetradecyl pyridyl bromide (TPB) was established (TPB) by flame atomic absorption spectrometry (FAAS). TPB was combined with cadmium tetrathocyanate to make neutral ion association compounds, which are extracted into xylene. The concentration of TPB can be indirectly determined by the absorption of cadmium in xylene layer with flame atomic absorption spectrometry. The linear range of TPB is 0.03-0.80 mg/L. The sensitivity is 0.0212 mg/L x 1%. When the method was applied to the analysis of waste water, the recovery is from 98% to 104% and the RSD is less than 9.16%. PMID- 10682595 TI - [Effects of low temperature storage on DNA migration assayed by single cell gel electrophoresis]. AB - V79 cells treated with potassium dichromate and control cells were stored at 4 degrees C, -20 degrees C, -80 degrees C and -196 degrees C. Single cell gel electrophoresis (SCGE) was carried out after preserving cells for 0, 4, 24, 48 and 168 hours. The results showed that there was no difference on DNA migration between cells processing immediately after trypsinization and cells preserving for 24 h at different low temperatures. Longer storage (48 h or 168 h) resulted in a significant increase in the length of DNA migration and decrease in cell viability. The results suggested that 4 degrees C is more convenient for less than 24 h storage without influence on DNA migration and -80 degrees C is better for longer storage to minimize the influence on DNA migration. PMID- 10682596 TI - [Effects of opium peptide system on the temperature regulation of mice under dry hot environment with different temperature]. AB - The Kunming mice were selected as subjects. Animals were allowed to move and drink freely. The Naloxone(0.02 mg/g) was injected by abdominal cavity. The ability to body temperature stable by mice under the environment with high temperature (41 degrees C, 43 degrees C) was observed. The results showed that after obstructing opium receptor by Naloxone, the ability mentioned above was reduced obviously. PMID- 10682597 TI - [Effect of dietary calcium on serum calcium and calmodulin activity of brain and hypothalamus in rats]. AB - The effect of low calcium (LC) diet supplemented with various amount of calcium on serum calcium and calmodulin of rats was studied. The LC diet was mainly composed of corn low in calcium. The calcium content of LC diet was only half of that of the stock diet. Seventy Wistar rats were divided into 7 groups by weight and sex. Results showed that serum calcium in LC group was low and calmodulin activity was also low. These parameters were improved while calcium was supplemented in LC diet. There is a significant dose-response relationship among diet calcium, serum calcium and growth. When the total calcium in diet was increased up to 1000 mg/kg, serum calcium level was closed to that of normal and higher than the serum calcium in LC group significantly. The study demonstrated that low dietary calcium, hypocalcemia and low calmodulin are associated. When total dietary calcium was up to 1000 mg/kg, the growth retardation was attenuated. PMID- 10682598 TI - [Study on the contamination level and the tolerable limit of mould and yeast in yoghurt]. AB - The results show that 67.33% of yoghurt samples collected from factories, supermarkets and retailers are contaminated by mould and yeast. The main problem of yoghurt product is yeast contamination (56.67%). The highest count of mould and yeast in the positive samples are 39,500 cft/ml and innumerable, respectively. There is a significant difference in the levels of contamination of mould and yeast between yoghurt filled in the glass bottles and that in the plastic bottles (P < 0.005). According to the results above, it is suggested that the tolerable limit of mould and yeast in yoghurt should be equal to or less than 50 cfu/ml. PMID- 10682599 TI - [Mutation of p53 gene in human fetal gastric mucosal cells by sterigmatocystin in vitro]. AB - Cell culture, flow cytometry and silver-staining PCR-SSCP methods were used to explore the effects of sterigmatocystin(ST) (1 mg/L and 3 mg/L) on carcinogenesis and mutation of tumor suppressor gene p53 in human fetal gastric mucosal cells in vitro. Four weeks after treated with ST, the cells showed vigorous growth and malignant transformation foci. Twenty-four weeks after ST treatment, the cells could form cellular colonies in soft agar(the mean colony number was 15 and 17 perdish for ST 1 mg/L and 3 mg/L groups respectively). Flow cytometric analysis showed that both proliferation indexes (PI) and the cellular DNA contents of ST treated cells were much higher than those of normal control. The DNA contents of ST treated cells were in DNA aneuploid range. Mutant p53 protein expression was also significantly higher in ST treated cells. Silver-staining PCR-SSCP analysis showed that abnormal electrophoretic migration bands could be seen at exon 8 of p53 gene in ST-treated groups 22 weeks after ST treatment, while no abnormal bands were found in control group. Thus, the results further confirmed the carcinogenic effects of ST on human fetal gastricmucosal cells. PMID- 10682600 TI - [Study on the antimutagenicity of curcumin]. AB - Curcumin is a coloring additive used in food, which has many biological functions. In order to provide basic information on its anticarcinogenicity, the antimutagenicity of curcumin was determined by Ames test and micronuclei test. The results indicated that curcumin (60-250 micrograms/plate) inhibited the revert mutant induced by 2, 7-diaminofluorene (2AF) with S-9 in TA98 and in TA100. The inhibitory rates are 29.5%-55.9% and 37.5%-59.1% respectively. Before the injection of cyclophosphamide (CP), mice were given with curcumin (60, 120, and 240 mg/kg) orally once a day for a week. The results revealed that curcumin reduced the micronuclei formation induced by CP. The inhibitory rates of curcumin are 43.7%, 56.9%, and 63.9% respectively, which are different from that of the positive control significantly. PMID- 10682601 TI - [Determination of taurine in foods by high performance liquid chromatography]. AB - Taurine (2-aminoethanesulfonic acid) was quantitated by reversed-phase liquid chromatography on a C18 resolve column (ZOBAX ODS) using a mobile phase of methyl alcohol-phosphate buffer (pH = 4.9). L-glutamine was used as the internal standard. Before separation, the sample was extracted with 0.2 mol/L sulfosalicylic acid solution and cleaned up with dual ion exchange column chromatography. The amino acid was derived with o-phthalaldehyde to form an amino acid adduct which was monitored at 340 nm by UV detector. The HPLC method for taurine analysis in foods showed the precision (relative standard deviation) is better than 5%. The recoveries are more than 95% and the detection limit is 12 ng of taurine. PMID- 10682602 TI - [Impact of maternal income on the nutrients intake of preschool children--a case study in 8 provinces of China]. AB - The data of this study are based on the project "China Health and Nutrition Survey" carried out in 8 provinces of China from 1989 to 1998 in collaboration with the University of North Carolina at Chapel Hill, USA. Totally, 1180 and 916 mother-child pairs aged 2-6 were investigated in 1991 and 1993 respectively. Descriptive and stratified analysis is applied to investigate the impact of maternal income on the nutrients intake of children. It is found that the nutrients intake of children in higher maternal income group is greater than that in lower maternal income group. After adjusting for family income, the results show that the more contribution of the mother's income to the family, the more nutrients intake of their children, especially in lower income families. It is illustrated that the income of mother may be more important for children's dietary status than that of other members of the family. PMID- 10682603 TI - [Survey on breakfast-eating behavior among residents in Beijing, Guangzhou and Shanghai]. AB - A survey of breakfast-eating behavior among residents in Beijing, Guangzhou and Shanghai was carried out in 1994-1995. The results showed that the breakfast eating rates in three cities were 74.8%-90.5%. Subjects aged below 35 years skipped breakfast more often. "Time limiting" in the morning became the main cause of breakfast-skipping. Nearly half of subjects in three cities got their breakfast out of home. Moreover, compared with two years ago, the present rates of eating outside increased at various levels in three cities. Steamed bun, bread and congee were main foods consumed at breakfast. In addition, 39% to 56% of subjects often drank milk. Through this survey, the authors got an all-round understanding on breakfast-eating rates, eating outside rates and variety of food consumed of residents in 3 cities. This survey will also provide a background for nutrition intervention and education in the future. PMID- 10682604 TI - [Comparison between selenomethionine and sodium selenite on action potentials of cultured myocardiocytes]. AB - The action potentials of ventricular myocardial cells were recorded with glass microelectrodes inside the cells from neonatal Wistar rats treated with selenomethionine and sodium selenite in concentrations of 1.0, 2.0, and 4.0 mg/L selenium. Both of selenomethionine and sodium selenite decreased the action potential parameters, such as action potential amplitude(APA), overshoot(OS), threshold potential(TP), maximum diastolic potential (MDP) and maximum rate of depolarization (Vmax). Selenomethionine decreased the action potential duration at 10%, 50% and 90% repolarization (APD10, APD50 and APD90), while sodium selenite prolonged APD10, APD50 and APD90 at dose of 4.0 mg/L selenium and decreased APD50 and APD90 at dose of 1.0 and 2.0 mg/L selenium. The results indicated that both sodium selenite and selenomethionine inhibit the transmembrane movement of Ca2- and sodium selenite also inhibits transmembrane movement of K+. PMID- 10682605 TI - [Study on certified reference material of germanium in Ganoderma lucidum]. AB - Analytical reference material of Ge in Ganoderma lucidum is designed and prepared for accurete analysis, monitoration and evaluation in trades of farming, forestry, medicine and food hygiene for Ge. It is used in technical training, technical assessing, monitoring, data arbitrating and analytic method verifing for professional supervisors. This reference material has been certified by graphitic oven atomic absorption spectrometry, hydride spectrophotometry, polarography, chemical separation spectrophotometry, atomic fluorescence method and x-ray fluorescence method. According to Grubb's law to judge the data of each group, it is confirmed that all of seven groups certified crude data are normal distribution by checking normality D. The arithmatic mean value of all data is 0.38 microgram/g. Standard deviation is 0.08 microgram/g. PMID- 10682606 TI - [Study on certified reference material of benzene, toluene, o-xylene in charcoal]. AB - This certified reference material (CRM) of benzene, toluene and o-xylene in charcoal is used for quility control and calibration of analytical instruments in the determination of benzene, toluene and o-xylene in air. Homogeneity of benzene, toluene and o-xylene concentrations in the CRM was testes by GC. The stability of the CRM was assessed for one year. The certified values are based on analysis made by a group of independent laboratoies using GC method. PMID- 10682607 TI - [Research and production of air cleaner for traveller train carriage]. AB - After the traveller train carriage is closed, the air pollution would be serious in the carriage. In order to control the air pollution, the air cleaning technology must be studied and the air cleaner for the carriage must be designed. The authors discussed. 1. Working out a technology scheme and main technology parameter for the air cleaner, and the structure design of the air cleaner. 2. Texting the function and performance of the air cleaner. 3. Investigating the effectiveness of the cleaner in same train sections. Thereby it will be confirmed that the air cleaner can improve the air environment in the carriage, and give convincingly security to the health of attendants and travellers. PMID- 10682608 TI - [Study on the relationship between deoxyribonucleoside triphosphate (dNTP) pools and cell transformation]. AB - Deoxyribonucleoside triphosphate (dNTP) pools were measured in normal BALB/c3T3 cells, transformation-treated cells and transformed cells with reverse-phase HPLC. The fluctuation of dNTP pools was similar after cells were treated with alkylating mutagens glycidyl methacrylate (GMA) and N-methyl-N'-nitro-N nitrosoguanidine (MNNG). The gap between (dGTP + dATP) pools and (dTTP + dCTP) pools was greatly intensified. The measurements also indicated that the dNTP pools in transformed cells were quite different from those in normal cells. The results suggest that dNTP pools may play an important role in cell transformation. PMID- 10682609 TI - [Study on inhibition and prevention of tumor and antioxidative effects of lithium carbonate in tumor bearing mice]. AB - Two Kinds of tumor-bearing mice (hepatoma H22 and sarcoma S180) were administered with lithium carbonate (Li2CO3) for 17 or 10 days (advanced and simultaneous administration), in order to observe the effects of prevention and treatment of Li2CO3 on malignant tumor, as well as the relationship between Li2CO3 and lipid peroxidation in tumor-bearing mice. Meanwhile, we compared the toxic and side effects of cyclophosphamide (CP) with that of Li2CO3. The results showed that Li2CO3 had no significant toxic or side effects with the suggested doses. In the tests of inhibition and prevention of tumor, Li2CO3 could significantly inhibit the grouth of the two kinds of tumor, and increase the activity of superoxide dismutage (SOD) and decrease the contents of Malonyldialdehyde (MDA). In addition, Li2CO3 had no effect on the white blood cells (WBC) and decreased the micronucleus frequency (MNF) in bone marrow polychromatic erythrocytes (PCE), while CP had definite effect of decreasing the WBC and increasing the MNF in the tumor-bearing mice. PMID- 10682610 TI - [Observation of inducing effect of five kinds of soft wood dust in micronucleus test]. AB - The authors investigated the micronucleus frequencies (MP) in peripheral lymphocytes in 584 workers exposed to five kinds of soft wood dust and found that all kinds of soft wood dust could increase the micronucleus frequency. As the control's 95% confidential upper limit > or = 0.4% is defined as the standard for positivity the micronucleus positive frequency (MPF) in the exposure group is much higher than that in the control group (P < 0.01). Meanwhile the increase of exposure years may increase the MP, and the time-response relationship is evident (r = 0.90, P < 0.01). For different exposures, the MPFs are: 36.2% for linden, 26.3% for fir. 15.7% for lacebark pine, 9.6% for birch and 2.1% for polar. The positive rate of linden and fir exposure are significantly higher than that of the control group (P < 0.01). Water extract of these five kinds of soft wood dust can increase the frequencies of micronucleus in polychromatic red blood cell in mice's sternum marrow, which also showed a dose-response. The correlation coefficients are: 0.86 for linden 0.6 for fir, 0.39 for lacebark pine, 0.76 for birch and 0.73 for polar. All but the lacebark pine have the P value less than 0.01 (P < 0.01). At the dose of 15 g/kg body weight, micronucleus frequency induced by linden was the highest (0.8%), next were fir (0.71%), birch (0.65%), lacebark pine (0.63%), and polar (0.43%). The results from animal test are consistent with those from the exposure workers. These results suggest that there are some substances in soft wood which can induce chromosome aberration. PMID- 10682611 TI - [Study on electroneuromyography of 175 workers exposed to carbon disulfide]. AB - A study on electroneuromyography of 175 workers exposed to carbon disulfide demonstrated that electroneuromyographic abnormalities of these workers were in accordance with characteristic alteration of axonal polyneuropathy. Electroneuromyography was an objective and sensitive method to detect the early damage of peripheral nerve caused by low-level exposure to carbon disulfide. The dose-response relationship between the exposure to carbon disulfide and the damage of peripheral nerve was demonstrated. PMID- 10682612 TI - [Study on method of sampling and analysis of trichloroaniline in the air]. AB - Trichloroaniline in the air was collected with 50% ethanol and separated with a column OV-17:OV-210 ECD as a detector. The best conditions of sampling and determining were selected through the orthogonal experimental design. The detectable limit was 0.01 mg/L. When the concentration of trichloroaniline in the air was 0.5-10 mg/L, the relative standard deviation (RSD) was 4.3%-3.7%. There was a linear relation within the range of 0.02-40 mg/L. The sampling efficiency was 99.1%-100%. The samples were stable for at least 15 days. It is proved that this method is accurate, fast, simple and sensitive. It is suitable for the determination of trichloroaniline in the air. PMID- 10682613 TI - [Role of programmed cell death in mediating arsenic-induced rat embryo anomalies]. AB - With a view to further investigating teratogenicity, embryotoxicity and teratogenic mechanism of arsenic on developing embryos, the relationship between apoptosis (programmed cell death, PCD) and arsenic-induced teratogenesis was studied during head-fold stage by using in situ 3'end-labeling of DNA and Nile blue sulfate (NBS) vital staining. Sprague-Dawley rats were intraperitoneally injected on gestation day 9.5 with doses of 0, 1, 4 and 8 mg/kg arsenic respectively, and killed at day 12, the results show that teratogenic incidence and mortality increased to 35.7% and 23.8% respectively, in apparent dose-effect relationship. In NBS staining experience, it was found that arsenic induced PCD in many areas, which related with abnormal development of brain, optic system, somite segmentation, limb bud and so on. In situ 3' end-labeling of DNA further supported above-mentioned results. The positive rate of apoptotic cell death was up to 55.6% (P < 0.05). This research suggested a positive correlation between abnormal PCD and teratogenesis. It may be seen that over-apoptosis is one of teratogenic mechanisms of arsenic. Moreover, our results show that necrosis plays some role in teratogenesis as well. PMID- 10682614 TI - [Study of ozonization effects on mineral water components]. AB - The disinfection effects of ozonization and its influences on chemical components of mineral water were investigated. The results showed that ozone at the level of 0.5 mg/L and with the exposure time of 5 minutes effectively destroyed bacteria in mineral water. High level ozone showed no strong influences on some beneficial components, such as strontium and metasilicate and on some main components, such as bicarbonate, hardness and alkalinity, but slightly elevated pH value. Ozonization reduced the contents of total dissolved solids and oxygen demand, and decomposed some reductive contaminants such as ammonia, cyanide and phenols. Ozonization will convert part of the bromide into hypobromite and bromate. PMID- 10682615 TI - [Effects of static magnetic fields on free radical metabolism of human body]. AB - The effects of static magnetic fields at 150 to 155 mT on the activities of superoxide dismutase (SOD) and the contents of maglonydiadehyde (MDA) in the serum and the activities of glutathione peroxidase (GSH-Px) in the whole blood were observed in the healthy volunteers. The results showed that the activities of SOD and GSH-Px were significantly increased, and the content of MDA was significantly decreased in short term at the observed magnitude of static magnetic fields. The results suggest that the function of the free radical metabolism of human body could be improved by static magnetic fields at that magnitude. PMID- 10682616 TI - [Detection of microcystins in source and tap water from a lake]. AB - The microcystins in source and tap water collected from seven waterworks around a lake in summer, 1996 were detected by enzyme-linked immunosorbent assay (ELISA). Microcystins were determined in the source water from all the seven waterworks, and the concentrations ranged from 280 to 35,300 ng/L. Low concentrations of microcystins were also detected in the tap water of three waterworks. The results suggest that microcystins in source water could not be removed completely by conventional adding bleaching powder treatment. PMID- 10682617 TI - [A rapid simple and reliable method for sequencing the targeted gene of shuttle vector pSP189]. AB - The authors established a simple, rapid and effective procedures for sequencing the targeted a-gene SupF TRNA of shuttle vector pSP189, which is very important and useful in molecular mechanism research of mutagens. In this method, double stranded plasmid DNA as template, prepared single-stranded DNA with alkali denaturation, applied r-32P-ATP for labelling 5'-end of primer, were used. A satisfactory result was achieved. PMID- 10682618 TI - [Efficacy of beta-carotene on lipid peroxidation induced by N nitrosodimethylamine in rats]. AB - The effect of beta-carotene (beta-C) on lipid peroxidation induced by N nitrosodimethylamine (NDMN) in rats was studied. Thirty clear conventional SD rats were randomly devided into 3 groups and were intragastricly given daily water (10 ml/kg). NDMN (1.75 mg/kg), and NDMN (1.75 mg/kg) plus beta-C (25 mg/kg) respectively. The results showed that: compared with group 1, the activities of SOD and GSH-Px in NDMN group were significantly decreased (P < 0.05), the values of MDA and ROOH increased (P < 0.05). The activities of SOD and GSH-Px in group 3 were significantly higher and the contents of ROOH and MDA were significantly lower than that of NDMN group (P < 0.05). These results suggested that free radicals and lipid peroxidation may be a carcinogenic path for NDMN and other nitrosamines. beta-C had ability to inhibit the lipid peroxidation induced by NDMN. PMID- 10682619 TI - [Scavenging effect of total flavonoids of lycium barbarum L on active oxygen radicals and inhibitory effects on heat output from L1210 cells]. AB - The scavenging effects of total flavonoids of Lycium barbarum L. (TFL) were studied by using ESR-spin trapping technique and the inhibitory effects on heat output of both polymorphonuclear leukocyte(PMN) respiration burst and L1210 cells were measured by using microcalorimetric technique. TFL (0-217 mg/L) could scavenge O2-. in xanthine/xanthine oxidase (Xan/XO)system, with scavenging rate of 0-51%. TFL(7.5-200 mg/L)could scavenge OH. produced in Fenton reaction and the scavenging rate is between 20% to 72%. Those effects were concentration dependent. Furthermore, TFL(0.56 g/L)could completely inhibit the heat output from PMA-stimulated PMN and TFL(1.0-5.0 g/L)could inhibit the heat output from L1210 cells. PMID- 10682620 TI - [Effect of zinc on cadmium-induced fetal damage]. AB - The effect of different zinc level on cadmium (Cd)-induced fetal damage was studied in pregnant rats. Seventy Waster rats were divided into five groups: zinc 30 mg/kg diet, zinc deficiency (3.2 mg/kg), zinc deficiency with Cd 2.0 mg/kg, zinc 30 mg with Cd 2.0 mg and zinc 227 mg with Cd 2.0 mg. The pregnant rats were killed on the 20 th day of gestation. Blood samples were taken and all the fetuses were examined. The result showed that zinc deficiency could induce fetus absorption or malformation, which were increased significantly in zinc dificient plus Cd rats. Fetuses can be protected if high dose of zinc (227 mg/kg) is given. The occurrence of fetus absorption and malformation decreased and survival increased in compared with the nonsupplemented group. The results suggested that fetal malformations can be induced by zinc deficiency, and being worse when Cd is added. Zinc supplementation can offset to some extent the toxic effect of Cd. PMID- 10682621 TI - [Behavioral-teratological effects of caffeine on mice]. AB - Pregnant ICR mice were exposed to caffeine at levels of 0, 4, 20 and 100 mg/kg during pregnancy. The behavioral-teratological tests indicated that the retardation of development, the obstruction of early reflexes and sensation and decline of learning ability existed in the offsprings of 20 and 100 mg/kg groups. The number of the dead fetus increased in the group of 100 mg/kg coffeine treated mice. No morphological changes were found. PMID- 10682622 TI - [Relationship between the toxigenicity and ribotype distribution of Pseudomonas cocovenenans subsp. farinofermentans]. AB - By rDNA fingerprinting technique, ribotype discrimination analysis was conducted for exploring the relationship between the toxigenicity and ribotype distribution of 42 strains of Pseudomonas cocovenenans subsp. farinofermentans. The results showed that 73.8 percent of the strains were toxigenic, which were distributed within the most ribotype and isolated from either food poisoning samples or normal food samples. The other strains were not toxigenic; which were isolated from normal food samples and distributed only within a few ribotypes. Since the most clones of P. cocovenenans subsp. farinofermentans were toxigenic, their potential hazard should be concerned. PMID- 10682623 TI - [Research on detection of type C2 staphylococcus enterotoxin in food with piezoelectric immunosensors]. AB - Using the biosensor of immobilization of antibody by Piezoectric Immunosensors (PEI) method, the concentration of antigen was determined. When the concentration of antigen was 10 micrograms/L-2 mg/L, the linear response was found. When the sensor was applied to detect type C2 staphylococcus enterotoxin in food, the method was proved with high specificity and selectivity. PMID- 10682624 TI - [Association of apoB levels and nutrition of coronary heart disease patients]. AB - Blood lipids and apo B levels were measured on 101 Coronary Heart Disease patients and 100 controls. The results showed that the apo B levels of patients were higher than that of controls. The average values were (877 +/- 129) mg/L and (800 +/- 95) mg/L respectively. It was found that there was close association among body mass index(BMI), ratio of waist circumference to hip circumference(WHR) and apo B levels. Besides, the results showed that dietary habit of "eating fat meat" might result in the increase of apo B concentration while the VC intake was negatively associated with serum apo B levels. PMID- 10682625 TI - [Catalytic spectrophotometric determination of trace manganese in theragran-M with Mn (II)-KIO4-NTA-coomassie brilliant blue G250]. AB - A new indicator reaction is developed on the basis of manganese catalyzed oxidation of coomassie brilliant blue G250 by potassium periodate with nitrilotriaetic acid as an activator. The detection limit for manganese is 6.76 x 10(-8) g/L. The linear range of determination is 0.02-0.30 microgram/25 mL. PMID- 10682626 TI - [Determination of serum copper by atomic absorption spectrometry sensitized with mixed surfactants]. AB - The effects of six kinds of surfactants were studied in the detection of copper in serum by atomic absorption spectrometry. The results demonstrated that the absorption decreased when polyvinyl alcohol was used alone or together with the other one, and the absorption increased when one or two of Tween-20, emulgent-OP, palmityl trimethyl ammonium bromide, arabicgum and sodium laurylsulfate were used. Tween-20 and sodium laurylsulfate used together are most effective. A method of determination of serum copper by atomic absorption spectrometry sensitized with mixture of Tween-20 and sodium laurylsulfate was established. The absorption increased by 50%. The relative standard deviation was 1.35%-2.11%. The recovery was 98.00%-98.13%. The detection limit was 0.0524 mumol/L. PMID- 10682627 TI - [Protein status and antioxidant capability of residents in endemic and nonendemic areas of Keshan disease(KD)]. AB - From a survey of residents in endemic and nonendemic areas of Keshan disease(KD), it was found that the plasma cystine, tryptophan and lysine as well as blood selenium content were lower, while the plasma lipid peroxide was higher in endemic areas than those in non-endemic areas. These results suggests that the selenium deficiency coexisting with cystine and tryptophan insufficiency decreases the antioxidant defense of the organism and may be one of the important factors for the development of KD. The role of sulfur-containing amino acid in antioxidant defense is discussed emphatically. PMID- 10682628 TI - [Comparative study on the retention effect of the inhaled mineral and coal dusts in rat lung]. AB - The pulmonary retention and physico-chemical characteristics of the dust are very important factors in the occurrence of pneumoconiosis. This paper reports a comparative study on the retention characteristics between the inhaled mineral dust and coal dust in rat lung. The dust quantities in the lungs and the mediastinal lymphatic nodes were determined by the formic acid digestion method. The results showed that the pulmonary retention quantities of the coal dust in rats were higher than those of the mineral dust on the third day but lower on the 90th day after exposure for two weeks. The results suggest that the mineral dust can deposit in the lung longer than the coal dust, and the lymphatic system is important for the pulmonary elimination of the dust. PMID- 10682629 TI - [Development of the certified reference material of iodine in lyophilized human urine]. AB - Urine samples from normal level and low level iodine districts were filtered, homogenized, dispensed, lyophilized and radio-sterilized. Homogeneity test, stability inspection and certification were conducted using a spectrophotometric method. The physical and chemical stability of the CRM (Certified reference material) were assessed for one year. The certified values are based on analysis made by five independent laboratories using three different principle methods. The certified values +/- uncertainties are as follows: low level is (121 +/- 14) micrograms/L, normal level is (201 +/- 11) micrograms/L. PMID- 10682630 TI - [Detection of K-ras oncogene mutations in human lung cancer by PCR-SSCP-DNA direct sequencing]. AB - K-ras oncogene mutations were detected with PCR-SSCP-DNA direct sequencing technique in 40 cases of lung cancer. The result of PCR-SSCP silver staining indicated that the mutational rate was 30% (12/40), all mutations were observed in lung adenocarcinoma and its mutational rate was 44% (12/27). The result of DNA direct sequencing showed that 11 of the 12 positive samples screened by PCR-SSCP had mutation and 90% of K-ras mutations were in codon 12. The mutation was mainly G-->T transversion and G-->A transition. The study suggested that SSCP silver staining analysis is very useful in screening large amount of samples simultaneously and PCR-SSCP-DNA direct sequencing method is quite efficient for the detection of oncogene mutation. PMID- 10682631 TI - [Study on the changes of heart-rate in lead-exposed workers]. AB - ECG examination and questionnaires survey were performed among lead-exposed workers in order to assess whether moderate occupational lead exposure may cause autonomic nervous system (ANS) dysfunction. The tests included; 1. heart-rate response to Valsalva manoeuvre (HR-V); 2. heart-rate variation during deep breathing (HR-DB); 3. immediate heart-rate response to immediate stand up (30:15 or Max:Min). The results did not show any evidence that lead exposure is related to ANS dysfunction. There was no significant difference between the exposed workers and controls who were with the same ages (P > 0.05). The important cause for ANS dysfunction was aging. PMID- 10682632 TI - [Evaluation on the effects of water defluoridation measures in China. Research Group Evaluation on the Effects of Water Defluoridation Measures in China]. AB - In order to find out the situation of management and application of water defluoridation measures, as well as the effects on the prevention of endemic fluorosis in 10 provinces and cities with heavy endemic fluorosis from the drinking water sources, 1960 water engineering projects accounting for about 10% of all projects were investigated. The authors applied uniform method, criteria and forms to carry out the retrospective investigation. The data were entered into the computer database and analyzed statistically. The results demonstrate that all the defluoridation projects have significant effectiveness on the prevention of endemic fluorosis. The concentrations of water fluoride were below 1 mg/L. The prevalences of dental fluorosis were 30%-40% and the main type was light dental fluorosis. But with the time prolonging, less attention has been paid in man. PMID- 10682633 TI - [Genetic epidemiological study of asthma in general]. AB - With theory and method of genetic epidemiology, data of 641 nuclear families on asthma were analysed, and the results showed that the history of parents asthma was one of risk factors about offsprings asthma, and the risk was 4.0-8.0 times higher (P < 0.01), compared with the one of offsprings of parents with no asthma. The effect of parents with asthma on the risk of sons was the same as one of daugters. The result declared that the airway responsiveness of parents did not relate to the one of offsprings. The results suggest that the genetic factors play an important role in the asthma. PMID- 10682634 TI - [Effects of low concentration ozone on the respiratory system of mice]. AB - Effects of low concentration of O3 on the respiratory system in mice were studied in the present study. Mice were exposed to 0.1, 0.25 and 1.0 mg/m3 of O3, respectively for 8 h per day for 3 days. Cilia in brochia, Lipid peroxide (LPO) in lung tissue and phagocytic function of macrophage in mice were examined. The results showed that there were cilia damage in the 1.0 mg/m3 group and swelling of mitochondrias as well as endoplasmic reticulums in the 0.25 mg/m3 group. But no damage was found in the group of 0.1 mg/m3. Furthermore, phagocytic rates in the 0.25 and 1.0 mg/m3 groups were significantly lower than that of control group (P < 0.01). No differences in LPO leads were found. PMID- 10682635 TI - [Data cleaning in the 1995-1996 national smoking prevalence survey]. AB - The authors introduced the procedure of data cleaning in the 1995-1996 national smoking prevalence survey, and the objectives, principles, procedures, quality evaluation and points for attention of data cleaning were summarized. Data cleaning must be carried out from the smallest investigative unit. The logical relationship among variables must be fully considered for the revision of incorrect values. In dealing with miss values, their roles in the statistical analyses must be evaluated. Comparing the differences in some important variables before and after cleaning is a useful method for assessing whether the representativeness of the data has been changed during the cleaning process. PMID- 10682636 TI - [Emergency measures on drinking water sanitation for mitigation of flood and waterlogging disasters]. AB - This article reports the emergency measures for mitigation during flood and waterlogging disasters to ensure drinking water sanitation and to prevent infectious disease outbreaks. Five preparatory and preventive measures for flood and waterlogging disasters include the construction of dual-purpose water supply installation for ordinary and disaster use, the storage of qualified technicians and materials (or their inventories), and the formulation of predetermination programme for disaster relief, ect. PMID- 10682637 TI - [The combined effects of MCYST and water organic pollutants on the co-induction of SHE cell transformation and expression of Ras P21 protein]. AB - The authors studied the combined carcinogenic activities of MCYST and organic pollutants in water of Dianshan Lake, using in vitro Syrian hamster embryo (SHE) cell transformation and immunohistochemical assays. The results showed that MCYST alone could not induce SHE cell transformation, but it increased the effects of low dose organic pollutants in cell transformation in a dose-dependent manner. In the cells isolated from type II transformed foci, ras P21 protein showed a positive immunohistochemical staining. It suggests MCYST may be a tumor promoter, which has a synergistic carcinogenic activity organic pollutants in water and the activation of ras oncogene may be one of the factors inducing malignant transformation. PMID- 10682638 TI - [Effects of selenium polysaccharide and sodium selenite on blood selenium concentration and liver cytochrome P450 monooxygenase system in rat]. AB - The effects of selenium polysaccharide and sodium selenite administered by single or repetitive intraperitoneal injection (i.p.) on blood selenium concentration, the activities of liver cytochrome P450, b5 as well as NAD(P)H cytochrome C reductase, glutathione S-transferase and glutathione were studied in rats. The biological effects of selenium polysaccharide and sodium selenite were also compared. The results indicated that the blood selenium concentration was increased rapidly and reached the peak in 2 hours followed by gradual decline after selenium polysaccharide and sodium selenite were i.p. injected at a dose of Se 0.6 mg/kg. The absorption and eliminating rates of Se from sodium selenite were faster than that from selenium polysaccharide. Administration of selenium polysaccharide and sodium selenite at a dose of 0.2 mg/kg by i.p. increased the blood selenium concentration to 2.6 and 2.1 times of those of control group, respectively, and the blood selenium concentration of selenium polysaccharide group was significantly higher than that of sodium selenite group (P < 0.05). The activities of liver cytochrome P450, b5 and GST were inhibited by selenium polysaccharide and sodium selenium in vivo and in vitro experiments. Those proteins were decreased to 57%, 70% and 62% of the control, respectively, by selenium polysaccharide which has particularly stronger effects on cytochrome P 450 monooxygenase system (P < 0.05). The two selenium compounds did not appear to affect the activity of NAD(P)H cytochrome C reductase. Both of the selenium polysaccharide and sodium selenite could enhance the activity of glutathion peroxidase significantly (P < 0.05). PMID- 10682639 TI - [The antioxidative mechanisms of tea polyphenols in inhibiting tumor promotion by TPA]. AB - In the mouse study, topical application of green tea polyphenols (GTP) significantly inhibited TPA-induced increasing of epidermal ornithine decarboxylase (ODC) and increased the activities of several antioxidant enzymes (CAT, GR and GST). In another in vitro study, when GTP was incubated with TPA and mice polymorphonuclear leukocytes (PMNs), TPA induced hydrogen peroxide formation was markedly suppressed with a dose-dependent relationship. The results suggest that the antioxidative effect of GTP may play an important role in inhibiting tumor promotion. PMID- 10682640 TI - [Short-term screening of anticarcinogenic ingredients of tea by cell biology assays]. AB - By using a panel of short term cell biology assays, several ingredients of tea (tea pigments, caffeine, tea polysaccharide, tea polyphenols tablet and mixed tea) were screened in order to investigate their anticarcinogenic effects. The cytokinesis block micronuclei test in V79 cells induced by mitomycin, the test of metabolic cooperation between V79 and M cells and the test of growth ability of Hela cells in soft agar were used in the screening. The results showed that the six kinds of tea ingredients tested were effective in the test involved in different stages of carcinogenesis, i.e. initiation, promotion and progression. The effects of mixed tea and tea pigments were the strongest among the ingredients tested. PMID- 10682641 TI - [Study on the molecular epidemiology characteristics of Pseudomonas cocovenenans subsp. farinofermentans isolated from China with rDNA fingerprinting]. AB - After the chromosomal DNA digestion by Bgl II and EcoRV, rDNA fingerprinting for Pseudomonas cocovenenans subsp. farinofermentans was made using biotin-labeled 16S + 23S rDNA as a probe. Fifty one strains of P. cocovenenans subsp. farinofermentans isolated from some areas of China were divided into 20 ribotyes, 52.9 percent of which were RT6, 9 and 2. In most cases, there were 2 or more ribotypes in each province or city. Though same ribotypes were isolated from different provinces, each ribotype existed in part of an area. The ribotype of RT6 was detected in samples from 4 incidents of food poisoning in Guizhou, Jiangsu and Hebei provinces, which indicated that these incidents may be caused by the same clone. With discrimination and cluster analysis of ribotypes, the rDNA fingerprinting may effectively contribute to analyzing the homogeneity of different strains, tracing the transmission route, and exploring the evolution and epidemiological characteristics of P. cocovenenans subsp. farinofermentans. PMID- 10682642 TI - [The method of removing methamidophos from contaminated vegetables]. AB - Since the massive food poisoning outbreak in 1987, imported vegetables contaminated with methamidophos continued to cause sporadic food poisoning outbreaks in Hong Kong. Despite various administrative measures to lower the risk of importing vegetables, it is evident that the occurrence of sporadic food poisoning outbreaks cannot be completely stemmed out. The education of the public on the effective ways to remove methamidophos from the contaminated vegetables was reckoned to the another preventive measure against food poisoning. A study was carried out to evaluate the effectiveness of various ways of treating the vegetables before consumption. In this study, the removal of methamidophos by simply washing with water at near room temperature was found to be a slow process. The concentration of methamidophos was reduced by about 65% after washing in water for an hour. Further washing did not improve the situation. The addition of detergents and various washing aids including potassium permanganate, hydrogen peroxide, sodium bicarbonate and vinegar did not greatly enhance the removal effectiveness. Among the various treatment procedures, soaking in hot water was the most effective way to remove methamidophos from vegetables. Less than 10% of the pesticide remained in the vegetables after soaking in boiling water for 1 minute. PMID- 10682643 TI - [The method for the determination of fluoride in the total diet study]. AB - This paper reports a microdiffusion method using fluoride-ion selective electrode for determination of fluoride contents in dietary samples. The method was suitable for the determination of fluoride in different dietary samples. It was a simple and fast method, and could be applied in large amount of sample analysis. The minimum detectable limit of this method was 1.0 microgram fluoride in 1.0 g dry dietary sample. The range of fluoride recovery and RSD in this dietary samples were 85.0%-107.0% and 4.5%-9.5% respectively. The fluoride content determined in the oyster tissue of NBS 1566 SRM was 87.9%-104.6% of certified SRM, and RSD was 6.5%. The RSD of fluoride determination in alcohol drinks was 3.1%. PMID- 10682644 TI - [Determination of copper, zinc, iron and calcium in wheat and maize and three nitrogen compounds in high and low risk areas of esophageal cancer]. AB - This study reports the concentrations of copper, zinc, iron and calcium in wheat and maize from high risk area Linzhou and low risk area Yuzhou of esophageal cancer and three nitrogen compounds of four types of drinking water in Linzhou. The results showed that the concentrations of zinc and calcium in wheat and maize from Linzhou were significantly lower than these from Yuzhou, the copper concentrations in grains were higher and the iron in maize was lower. The nitrate N concentrations in four types of drinking water from Linzhou were below the national standard but the nitrite and NH3-N concentrations were higher than the national limits. The results suggest that the low concentrations of zinc, iron and calcium and high concentrations of copper in grains as well as the high concentrations of nitrite and NH3-N may be related to the etiology of esophageal cancer. PMID- 10682645 TI - [Successful treatment of an elderly woman after stubborn resistance]. AB - A 72-year-old depressed woman was admitted by court order after a long history of ?successful' resistance to any treatment, both at home and in a psychiatric hospital. The nature of her disorder had not been recognised and she was diagnosed elsewhere as suffering from factitious disorder, probably based on intense countertransference. The diagnosis of depression with mood-congruent psychotic features was made. After several unsuccessful combination treatments and refusal of electroshock therapy, she finally responded to combination therapy with tranylcypromine, lithium carbonate and clozapine. Dutch medicolegal regulations need not be an impediment in such cases, intercollegiate consultation is most useful and strict adherence to therapy guidelines is beneficial. PMID- 10682646 TI - [The operating room of the future]. AB - Advances in computer technology will revolutionize surgical techniques in the next decade. The operating room (OR) of the future will be connected with a laboratory where clinical specialists and researchers prepare image-guided interventions and explore the possibilities of these techniques. The virtual reality is linked to the actual situation in the OR with the aid of navigation instruments. During complicated operations the images prepared preoperatively will be corrected during the operation on the basis of the information obtained peroperatively. MRI currently offers maximal possibilities for image-guided surgery of soft tissues. Simpler techniques such as fluoroscopy and echography will become increasingly integrated in computer-assisted peroperative navigation. The development of medical robot systems will make possible microsurgical procedures by the endoscopic route. Tele-manipulation systems will also play a part in the training of surgeons. Design and construction of the OR will be adapted to the surgical technology, and include an information and control unit where preoperative and peroperative data come together and from where the surgeon operates the instruments. Concepts for the future OR should be regularly adjusted to allow for new surgical technology. PMID- 10682647 TI - [Pharmacotherapy of patients with (early) rheumatoid arthritis]. AB - As soon as the diagnosis 'early rheumatoid arthritis (RA)' is made, a disease modifying antirheumatic drug (DMARD) should be prescribed without delay. Methotrexate in dosages up to 30 mg once weekly is being used more frequently than in the past, also in early RA. Combination therapy with DMARDs is indicated in case of insufficient effect of a single DMARD. Combinations with methotrexate appear to be especially effective, like methotrexate and cyclosporin. A novel effective DMARD is leflunomide. In the near future promising biologicals will probably be applied in clinical daily practice, presumably in combination with conventional DMARDs. New non-steroidal anti-inflammatory drugs (NSAIDs) have been developed that are probably safer than conventional NSAIDs. If the recent finding that glucocorticoids are able to inhibit joint damage in (early) RA will be confirmed, prednisone might be used more often in (early) RA. Bone marrow transplantation in RA is still experimental. PMID- 10682648 TI - [Physical diagnosis--percussion and palpation of the spleen]. AB - The accuracy of physical examination of the spleen was investigated in the literature. Ultrasonography or scintigraphy was used to test the findings at physical examination. Physical examination has a low sensitivity, but a reasonably good specificity. The interobserver variability is rather high. In general, palpation is more sensitive and specific than percussion. The findings of percussion improve the accuracy of palpation and the combination has a high specificity, of approximately 90%. The two should therefore be used in conjunction. The sensitivity is much lower although it is greatly influenced by the degree of splenic enlargement and the leanness of the patient. PMID- 10682649 TI - [Moroccans' opinions about general practitioners: analyzing reasons for consultation]. AB - OBJECTIVE: To determine whether there is a difference in the extent to which the GP succeeds in establishing the reasons for consultation of Moroccans and those of the Dutch; and whether the opinion of Moroccans about the GP's consultation differs from that of the Dutch. DESIGN: Analysis of patient interviews and GP's consultations. METHOD: In 11 general practices in Amsterdam and The Hague in May 1997, 50 Moroccan adults and 50 Dutch individuals were asked for their reasons to attend before the consultation and in the mother language; a distinction was made between the actual complaint and the expectations with regard to the consultation. The GPs recorded these data after every consultation. The complaints were coded by organ system and by nature of complaint, following which agreement of the assessments was scored on a scale ranging from 0 to 100. RESULTS: Both groups comprised 20 men and 30 women. The mean age of the Moroccans was 38.6 years (SD: 13.8), that of the Dutch 56.4 years (SD: 16.7). The GPs established complaints of Moroccan patients not as well as those of Dutch patients (score: 73.9 versus 87.3); the difference was more pronounced where patients with only elementary education were concerned (67.0 as against 86.1). The GPs were able to establish the expectations with regard to the consultation nearly as often for the Moroccan as for the Dutch patients (58.5 versus 55.9). Moreover, the Moroccans were as positive about the course of the consultation as the Dutch. Except for communication problems among the lower educated, none of the problems indicated appeared to be experienced more often by the Moroccans than by the Dutch. CONCLUSION: A large part of the complaints presented by lower educated Moroccan patients were interpreted differently by the GP. For Moroccans with a higher education, the care was comparable with that among the Dutch. PMID- 10682650 TI - [Fewer x-rays while maintaining quality of clinical care using clinical protocols for physical diagnosis of ankle injuries]. AB - OBJECTIVE: To determine whether it is possible to decrease the number of X-rays in acute ankle injury while keeping the health care constant, using a scoring system. DESIGN: Prospective. METHOD: Patients presenting in the emergency department of the University Hospital Utrecht (AZU), the Netherlands, over a one year period of time with acute ankle injuries were subjected to a thorough physical examination based on a scoring system developed at Leiden University Hospital. The score was calculated and X-ray examination was indicated when this score was > or = 8 points. Radiological investigation or telephone interviews six weeks after injury achieved verification of the clinically relevant ankle fractures. Specificity and sensitivity were calculated from every possible cut off point and drawn in a 'receiver operating characteristics' (ROC) curve. RESULTS: Of the 514 patients included 81 patients had a score of 8 or higher and 24 of them had a clinically relevant fracture. In 34 patients an ankle X-ray was made although their score was < 8 points. The positive and negative predictive values of the system were 30% (95% confidence interval (95% CI): 20-41) and 99% (95% CI: 97-100) respectively. The score yielded an area under the ROC curve of 91% (95% CI: 84-98). A cut-off point of 8 led to a reduction of X-rays by 60% (using the 'Ottawa ankle rules' the decrease in this population would have been 28%). On the other hand, 5 clinically relevant fractures were missed. CONCLUSION: Radiological examination in patients wit acute ankle complaints was reduced while health care remained almost constant. In the AZU, a decision was made for a major reduction in X-rays while accepting that some fractures would be missed. PMID- 10682651 TI - [Systemic capillary leak syndrome]. AB - A man aged 61 had recurrent attacks of severe shock. The episodes were preceded by symptoms such as a runny nose, epigastric discomfort with nausea, vertigo, orthostatism and sometimes light fever. During the attacks there were marked hypotension, a strong rise of the haematocrit, a decrease of the protein and albumin concentrations in the blood and prerenal kidney failure. In addition, there was a paraprotein, type IgG-kappa. The shock every time responded rapidly to intravenous administration of fluid and was followed by a period of substantial polyuria. The pattern was characteristic of systemic capillary leak syndrome, a rare but frequently fatal disease characterized by episodes of unexplained extravasation of plasma. The aetiology and pathogenesis are unknown. Attacks are suppressed by supportive therapy (administration of fluids, inotropics) and future attacks may be prevented by the intake of terbutaline and theophylline. The systemic capillary leak syndrome should be considered in the differential diagnosis of idiopathic and anaphylactic shock. PMID- 10682652 TI - [Do medical schools still train doctors?]. AB - In practice, medical finals are not final and a supplementary training is necessary before medicine may be practised independently. Social factors and an ongoing evolution of medical science prompt reconsideration of the structure, content and duration of the training of doctors and specialists. This was the subject of a meeting of this Journal. One possibility of differentiation in the basic training is an early subdivision into care physicians, clinical specialists and health physicians. In the training of social medical officers one of the factors to be taken into account is the influence of principals. For GP's, postgraduate training is increasingly important because of social and other developments. The training of non-surgical specialists can be made shorter since a significant proportion of the time in the present training is devoted to areas requiring special attention. The training of surgical specialists could be shortened by introducing a training programme that is independent of the procedure. Responsibility for the total care of the patient will be borne by the specialists jointly. PMID- 10682653 TI - [General practitioner: acrobat or drug store clerk?]. PMID- 10682654 TI - [Family doctor: acrobat or drugstore clerk?]. PMID- 10682655 TI - [Family doctor: acrobat or drugstore clerk?]. PMID- 10682657 TI - Hypermethylation of ribosomal DNA in human breast carcinoma. AB - We examined the methylation status of the transcribed domain of ribosomal DNA (rDNA) in 58 patients with breast cancer. The mean percent of methylation was significantly higher in breast tumours than that of normal control samples (P < 0.0001). This increased rDNA methylation was associated with oestrogen receptor non-expression (P < 0.0273) and with moderately or poorly differentiated tumours as compared to well differentiated tumours (P < 0.0475). Our results suggest that rDNA can be a useful marker for monitoring aberrant methylation during breast tumour progression. PMID- 10682656 TI - Oestrogen and growth factor cross-talk and endocrine insensitivity and acquired resistance in breast cancer. PMID- 10682658 TI - 17Beta-hydroxysteroid dehydrogenase type 1 and type 2 in human breast carcinoma: a correlation to clinicopathological parameters. AB - The expression of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) type 1 and type 2 was examined immunohistochemically in 111 invasive ductal carcinomas, and correlated with various clinicopathological parameters. This study investigates local regulatory mechanisms of oestrogens in human breast carcinoma. 17Beta-HSD type 1 was immunolocalized in carcinoma cells of 68 out of 111 invasive ductal carcinoma cases (61.3%). 17Beta-HSD type 2 immunoreactivity was not detected in all cases examined. A significant inverse correlation was observed between the immunohistochemical expression of 17beta-HSD type 1 and histological grade of the carcinoma (P < 0.02). There was a significant correlation between 17beta-HSD type 1 and oestrogen receptor (ER) labelling index (LI) (P < 0.05). In addition, carcinoma cells expressing immunoreactive 17beta-HSD type 1 were frequently positive for ER. 17Beta-HSD type 1 was also correlated with progesterone receptor (PR) LI (P < 0.05). There was a significant inverse correlation between 17beta HSD type 1 and Ki-67 LI (P < 0.0001). No significant correlations were detected between 17beta-HSD type 1 and other clinicopathological parameters, including patient age, menopausal status, stage, tumour size, lymph node status and prognosis. This study suggests that 17beta-HSD type 1 plays an important role in the regulation of in situ oestradiol production in hormone-dependent breast carcinomas. PMID- 10682659 TI - Idarubicinol myelotoxicity: a comparison of in vitro data with clinical outcome in patients treated with high-dose idarubicin. AB - We evaluated in vitro the toxicity of idarubicin and its active metabolite idarubicinol on haematopoietic progenitors, using human umbilical cord blood and peripheral blood progenitors to obtain dose-response curves. We treated 16 patients with poor prognosis lymphoma in a phase I-II trial of high-dose idarubicin and melphalan and investigated if idarubicinol persisting in patients' plasma at the time of transplantation (day 0), on day +1 and +2 could result in an inhibition of infused progenitors. Colony inhibition was correlated with pharmacokinetic data and with the time of patients' engraftment. Plasma samples obtained before idarubicin treatment demonstrated a colony-stimulating effect, increasing the cloning efficiency by 72%. The inhibitory activity on colony forming unit granulocyte-macrophage (CFU-GM) of patients' plasma collected on the day of transplantation was lower than expected from dose-response curves (21% measured vs 70% expected). The time to patients' WBC and PLT recovery correlated with the amount of CD34+ cells reinfused and, to a lesser extent, with the colony inhibiting effect of patients' plasma. The correlation between idarubicinol concentration and CFU-GM inhibition was not significant. These data suggest that plasma drug concentration on the day of stem cell reinfusion may overestimate the toxicity of residual anthracyclines to the transplanted cells. PMID- 10682660 TI - A unified definition of clinical anthracycline resistance breast cancer. AB - The purpose of the study was to determine the response rates (RR) and duration to second- and third-line chemotherapy programmes in patients with anthracycline resistant breast cancer, utilizing various definitions of anthracycline resistance. This was a retrospective analysis performed on 1335 patients with metastatic breast cancer who participated in consecutive clinical trials of first line, anthracycline-containing combination chemotherapy (ACCC) at the University of Texas MD Anderson Cancer Center between July 1973 and April 1980. Anthracycline-resistant groups were identified using definitions of anthracycline resistance found in the literature: progressive disease as best response to ACCC (Group 1, n = 56 patients); progressive disease while receiving ACCC after an intervening response to the drug (Group 2, n = 84); progressive disease within 6 months of last dose of ACCC (Group 3, n = 233); and progressive disease within 12 months of last dose of ACCC (Group 4, n = 272). Second- and third-line therapies administered to these patients included methotrexate, doxorubicin, mitoxantrone, bisantrene, vinblastine, vindesine, melphalan, mitomycin, cisplatin, etoposide and others, but not taxanes. The distribution of patients' characteristics was similar between the four groups, as was the use of second- and third-line regimens. Response rate (RR) to second-line chemotherapy were 5% and 7.7% for Group 1 and Group 2 respectively. In contrast, RR to second-line chemotherapy were 21.6% and 15% for Group 3 and 4. The differences in response rate between the combination of Groups 1 and 2 and Groups 3 or 4 were significant (P = 0.005 and P = 0.04 respectively). These results indicate that strictly defined anthracycline resistance as defined in Groups 1 and 2 is associated with resistance to many other cytotoxic drugs. The definitions used in Groups 3 and 4 include many patients with responsive tumours, and a more favourable prognosis. PMID- 10682661 TI - Intron splice acceptor site polymorphism in the hMSH2 gene in sporadic and familial colorectal cancer. AB - A polymorphism in hMSH2 gene has been associated with an increased susceptibility to develop colorectal cancer (CRC). Here we show that it is a genetic risk factor for CRC in the Spanish population. However, its presence does not apparently affect hMSH2 function. PMID- 10682662 TI - The prevalence of BRCA1 mutations in Chinese patients with early onset breast cancer and affected relatives. AB - The purpose of this study was to determine the prevalence of BRCA1 mutations in Chinese breast cancer patients in Singapore. BRCA1 analysis was conducted in consecutive patients with breast cancer before the age of 40 years (76 women), or whose relatives had breast or ovarian cancer (16 women). Ten patients had both early onset breast cancer and affected relatives. Genomic DNA from peripheral mononuclear blood cells was studied by using the protein transcription translation assay (exon 11) and single-strand conformational polymorphism, with subsequent DNA sequencing. All six disease-causing mutations occurred in women under 40 years (8.6%) with three occurring in patients under 35 years (three out of 22 patients, 13.6%). Mis-sense mutations of unknown significance were found in three patients. Two of the ten women with affected relatives under 40 years had BRCA1 mutations. The prevalence of BRCA1 mutations in Chinese patients with early onset breast cancer is similar to that observed in Caucasian women. Most Chinese patients with affected relatives were not carriers of BRCA1 mutations. PMID- 10682663 TI - Multiple deleted regions on the long arm of chromosome 6 in astrocytic tumours. AB - Chromosome 6 deletions are common in human neoplasms including gliomas. In order to study the frequency and identify commonly deleted regions of chromosome 6 in astrocytomas, 159 tumours (106 glioblastomas, 39 anaplastic astrocytomas and 14 astrocytomas malignancy grade II) were analysed using 31 microsatellite markers that span the chromosome. Ninety-five per cent of cases with allelic losses had losses affecting 6q. Allelic losses were infrequent in astrocytomas malignancy grade II (14%) but more usual in anaplastic astrocytomas (38%) and glioblastomas (37%). Evidence for clonal heterogeneity in the astrocytomas and anaplastic astrocytomas was frequently observed (i.e. co-existence of subpopulations with and without chromosome 6 deletions). Clonal heterogeneity was less common in glioblastomas. Five commonly deleted regions were identified on 6q. These observations suggest that a number of tumour suppressor genes are located on 6q and that these genes may be involved in the progression of astrocytic tumours. PMID- 10682664 TI - Analysis of the insertion/deletion polymorphism of the human angiotensin converting enzyme (ACE) gene in patients with renal cancer. AB - The angiotensin I-converting enzyme (ACE) contains an insertion/deletion (I/D) polymorphism, with the DD genotype associated with benign renal diseases. The distribution frequencies of the D and I alleles, and the DD, DI and II genotypes were determined in DNA extracted from kidney tissues of 58 renal cancer patients. The observed frequencies in patients who develop renal cancer was not significantly different than the normal population. PMID- 10682665 TI - Identification of a founder BRCA2 mutation in Sardinia. AB - Sardinian population can be instrumental in defining the molecular basis of cancer, using the identity-by-descent method. We selected seven Sardinian breast cancer families originating from the northern-central part of the island with multiple affected members in different generations. We genotyped 106 members of the seven families and 20 control nuclear families with markers flanking BRCA2 locus at 13q12-q13. The detection of a common haplotype shared by four out of seven families (60%) suggests the presence of a founder BRCA2 mutation. Direct sequencing of BRCA2 coding exons of patients carrying the shared haplotype, allowed the identification of a 'frame-shift' mutation at codon 2867 (8765delAG), causing a premature termination-codon. This mutation was found in breast cancer patients as well as one prostate and one bladder cancer patient with shared haplotype. We then investigated the frequency of 8765delAG in the Sardinian breast cancer population by analysing 270 paraffin-embedded normal tissue samples from breast cancer patients. Five patients (1.7%) were found to be positive for the 8765delAG mutation. Discovery of a founder mutation in Sardinia through the identity-by-descent method demonstrates that this approach can be applied successfully to find mutations either for breast cancer or for other types of tumours. PMID- 10682666 TI - Thymidilate synthase and p53 primary tumour expression as predictive factors for advanced colorectal cancer patients. AB - The purpose of this work was to analyse the ability of p53 and thymidilate synthase (TS) primary tumour expression to retrospectively predict clinical response to chemotherapy and long-term prognosis in patients with advanced colorectal cancers homogeneously treated by methotrexate (MTX)-modulated-5 fluorouracil (5-FU-FA). A total of 108 advanced colorectal cancer patients entered the present retrospective study. Immunohistochemical p53 (pAb 1801 mAb) and TS (TS106 mAb) expression on formalin-fixed paraffin-embedded primary tumour specimens was related to probability of clinical response to chemotherapy, time to progression and overall survival. p53 was expressed in 53/108 (49%) tumours, while 54/108 (50%) showed TS immunostaining. No relationship was demonstrated between p53 positivity and clinical response to chemotherapy (objective response (OR): 20% vs 23%, in p53+ and p53- cases respectively) or overall survival. Percent of OR was significantly higher in TS-negative with respect to TS-positive tumours (30% vs 15% respectively; P < 0.04); simultaneous analysis of TS and p53 indicated 7% OR for p53-positive/TS-positive tumours vs 46% for p53-positive/TS negative tumours (P < 0.03). Logistic regression analysis confirmed a significant association between TS tumour status and clinical response to chemotherapy (hazard ratio (HR): 2.91; 95% confidence interval (CI) 8.34-1.01; two-sided P < 0.05). A multivariate analysis of overall survival showed that only a small number of metastatic sites was statistically relevant (HR 1.89; 95% CI 2.85-1.26; two-sided P < 0.03). Our study suggests that immunohistochemical expression of p53 and TS could assist the clinician in predicting response of colorectal cancer patients to modulated MTX-5-FU therapy. PMID- 10682667 TI - Lobular carcinoma in situ of the breast is not caused by constitutional mutations in the E-cadherin gene. AB - Lobular carcinoma in situ (LCIS) is an unusual histological pattern of non invasive neoplastic disease of the breast occurring predominantly in women aged between 40 and 50 years. LCIS is frequently multicentric and bilateral, and there is evidence that it is associated with an elevated familial risk of breast cancer. Although women with LCIS suffer an increased risk of invasive breast disease, this risk is moderate suggesting that LCIS may result from mutation of a gene or genes conferring a high risk of LCIS, but a lower risk of invasive breast cancer. The high frequency of somatic mutations in E-cadherin in LCIS, coupled with recent reports that germline mutations in this gene can predispose to diffuse gastric cancer, raised the possibility that constitutional E-cadherin mutations may confer susceptibility to LCIS. In order to explore this possibility we have examined a series of 65 LCIS patients for germline E-cadherin mutations. Four polymorphisms were detected but no pathogenic mutations were identified. The results indicate that E-cadherin is unlikely to act as a susceptibility gene for LCIS. PMID- 10682668 TI - Loss of DCC gene expression during ovarian tumorigenesis: relation to tumour differentiation and progression. AB - To clarify the possible role of DCC gene alteration in ovarian neoplasias, we immunohistochemically investigated 124 carcinomas, as well as 55 cystadenomas and 41 low malignant potential (LMP) tumours and compared the results with those for p53 protein expression, clinicopathological factors and survival. A combination of the reverse transcription polymerase chain reaction (RT-PCR) and Southern blot hybridization (SBH) for DCC mRNA levels was also carried out on 26 malignant, five LMP, eight benign and seven normal ovarian samples. Significantly decreased levels of overall DCC values in carcinomas compared with benign and LMP lesions were revealed by both immunohistochemical and RT-PCR/SBH assays. Similar findings were also noted when subdivision was into serous and mucinous categories. In carcinomas, reduction or loss of DCC expression was significantly related to the serous phenotype (serous vs non-serous, P < 0.0001), a high histological grade (grade 1 vs 2 or 3, P < 0.02) and a more advanced stage (FIGO stage I vs II/III/IV, P = 0.0083), while no association was noted with survival. Although p53 immunopositivity demonstrated significant stepwise increase from benign through to malignant lesions, there was no clear association with DCC score values. The results indicated that impaired DCC expression may play an important role in ovarian tumorigenesis. In ovarian carcinomas, the altered expression is closely linked with tumour differentiation and progression. PMID- 10682669 TI - Spontaneous apoptosis in ovarian carcinomas: a positive association with p53 gene mutation is dependent on growth fraction. AB - Changes in cell survival contribute to tumour development, influence tumour biology and its response to chemotherapy. p53 gene alterations should negatively affect apoptosis by impaired p53-dependent apoptotic response. We looked for associations between spontaneous apoptosis, p53 gene mutation, p53 protein accumulation, growth fraction, bcl-2 expression and histological parameters in 64 ovarian, four tubal and three peritoneal carcinomas. Apoptotic cells were detected with the TUNEL method. p53 gene variants were detected by the single strand conformation polymorphism and were sequenced directly. P53, Ki-67 and bcl 2 protein expressions were detected immunohistochemically. A weighed multiple logistic regression model was applied. Apoptotic index (AI) ranged 0.02-0.18 (mean 0.11); proliferation index (PI) ranged 3-90% (mean 54%). p53 gene mutations were present in 51, p53 protein accumulation in 46, and diffuse bcl-2 expression in 29 of 71 tumours. The AI was positively associated with the presence of p53 gene mutation (P = 0.011). However, the PI included into the analysis did positively influence the AI (P = 0.02) and diminished the association with p53 gene mutation (P = 0.082). The AI was negatively associated with good histological differentiation (P = 0.0006), the serous tumour type (P = 0.002), and diffuse bcl-2 expression (P = 0.025). Strong bcl-2 expression was associated with endometrioid tumour type (P = 0.002). FIGO stage and p53 protein accumulation were the only parameters that influenced overall survival time. Thus, our results suggest that histological tumour type and grade are major determinants of spontaneous apoptosis in ovarian carcinomas; p53 alterations do not adversely but rather positively affect spontaneous apoptosis by increasing growth fraction. This, in turn, suggests p53-independency of spontaneous apoptosis in ovarian carcinomas. PMID- 10682670 TI - Tumour prothymosin alpha content, a potential prognostic marker for primary breast cancer. AB - In a previous report we suggested that the estimation of prothymosin alpha (PTA) levels in primary breast tumours might be used to identify breast cancer patients at high risk for distant metastasis (Dominguez F et al (1993) Eur J Cancer 29A: 893-897). Here the role of tumour PTA levels as predictor was investigated with respect to both disease-free survival (DFS) and survival. Tumours were obtained from a series of 210 consecutive female patients with ductal carcinoma who underwent surgery at the Hospital Xeral de Galicia (Santiago de Compostela, Spain). Characteristics including PTA tumour levels, number of positive axillary nodes, patient's age at surgery and tumour histological grade were significantly associated with DFS and survival, as determined by univariate analysis. Patients with tumours with low or moderate PTA levels demonstrated a statistically decreased rate of tumour recurrence and a statistically significant increased overall survival in comparison with those whose tumours had high PTA levels. Patient's relative risk of dying was 2.1 times greater for tumours with high PTA levels than for those tumours with low or moderate PTA levels. In conclusion, these data support the hypothesis that tumour high PTA levels is associated with a worse outcome. PMID- 10682671 TI - Angiogenic switch occurs late in squamous cell carcinomas of human skin. AB - Angiogenesis is a crucial event in carcinogenesis and its onset has been associated with premalignant tumour stages. In order to elucidate the significance of angiogenesis in different stages of epithelial skin tumours, we analysed the vessel density in ten normal skin samples, 14 actinic keratosis (AK), 12 hypertrophic AKs, and in nine early- and 16 late-stage squamous cell carcinomas (SCCs). Mean vascular density was quantitated by counting the number of CD 31-immunostained blood vessels and by morphometric assessment of stained vessel area by computer-assisted image analysis. The results from both methods were well correlated. Mean vascular density was similar in normal dermis and in AK, and only slightly elevated in hypertrophic AKs and early SCC stages (tumour thickness < 2 mm). Only late-stage SCCs infiltrating the subcutis exhibited a significant increase in vascularization. Vessel density was independent of tumour localization, degree of proliferation and inflammatory cell infiltration. Furthermore, tumour vascularization was not correlated with the expression of vascular endothelial growth factor, a major angiogenic factor, as revealed by in situ hybridization and immunohistochemistry. The restriction of enhanced vascularization to increased tumour thickness may be a major reason for the rather low metastatic spread of cutaneous SCCs. PMID- 10682672 TI - Telomere length and telomerase activity in malignant lymphomas at diagnosis and relapse. AB - Telomere length maintenance, in the vast majority of cases executed by telomerase, is a prerequisite for long-term proliferation. Most malignant tumours, including lymphomas, are telomerase-positive and this activity is a potential target for future therapeutic interventions since inhibition of telomerase has been shown to result in telomere shortening and cell death in vitro. One prerequisite for the suitability of anti-telomerase drugs in treating cancer is that tumours exhibit shortened telomeres compared to telomerase positive stem cells. A scenario is envisioned where the tumour burden is reduced using conventional therapy whereafter remaining tumour cells are treated with telomerase inhibitors. In evaluating the realism of such an approach it is essential to know the effects on telomere status by traditional therapeutic regimens. We have studied the telomere lengths in 47 diagnostic lymphomas and a significant telomere shortening was observed compared to benign lymphoid tissues. In addition, telomere length and telomerase activity were studied in consecutive samples from patients with relapsing non-Hodgkin's lymphomas. Shortened, unchanged and elongated telomere lengths were observed in the relapse samples. The telomere length alterations found in the relapsing lymphomas appeared to be independent of telomerase and rather represented clonal selection random at the telomere length level. These data indicate that anti-telomerase therapy would be suitable in only a fraction of malignant lymphomas. PMID- 10682673 TI - Early evaluation of tumour metabolic response using [18F]fluorodeoxyglucose and positron emission tomography: a pilot study following the phase II chemotherapy schedule for temozolomide in recurrent high-grade gliomas. AB - Quantitation of metabolic changes in tumours may provide an objective measure of clinical and subclinical response to anticancer therapy. This pilot study assesses the value of quantitation of metabolic rate of glucose (MRGlu) measured in mmol min(-1) ml(-1) to assess early subclinical response to therapy in a relatively non-responsive tumour. Nine patients receiving the CRC Phase II study schedule of temozolomide were assessed with [18F]fluorodeoxyglucose ([18F]FDG) dynamic positron emission tomography (PET) scans prior to and 14 days after treatment with temozolomide given as 750-1000 mg m(-2) over 5 days every 28 days. Tumour MRGlu was calculated and compared with objective response at 8 weeks. Pretreatment MRGlu was higher in responders than non-responders. The responding patient group had a greater than 25% reduction in MRGlu in regions of high focal tumour uptake (HFU). Whole tumour changes in MRGlu did not correlate with response. Percentage change in HFU standardized uptake value (SUV) did discriminate the responding from the non-responding patients, but not as well as with MRGlu. Large differences also occurred in the normal brain SUV following treatment. Thus, MRGlu appeared to be a more sensitive discriminator of response than the simplified static SUV analysis. Changes in MRGlu may reflect the degree of cell kill following chemotherapy and so may provide an objective, quantitative subclinical measure of response to therapy. PMID- 10682674 TI - Diagnosis of persistent ovarian carcinoma with three-step immunoscintigraphy. AB - The diagnosis of recurrent ovarian carcinoma is usually determined at surgical re exploration since the main non-invasive diagnostic tests have low accuracy. It would be desirable to have a high accuracy non-invasive diagnostic procedure. With this aim, we have assessed the utility of three-step immunoscintigraphy. Thirty patients were intravenously injected with biotinylated monoclonal antibodies MOv18 and B72.3, followed by avidin-streptavidin injection and finally by 111In-biotin. Tumour recurrences were imaged 2 h post radioactivity injection. All patients underwent surgical re-exploration 3-4 days after immunoscintigraphy; the presence of tumour in the area of immunoscintigraphic uptake was evaluated in the biopsied material. Twenty-one patients studied were true-positive, five were true-negative, four were false-positive and none was false-negative. The diagnostic accuracy, positive predictive value and negative predictive value were 87%, 84% and 100% respectively. If these findings are confirmed in a larger number of patients, we expect immunoscintigraphy to be introduced as a cost effective procedure in the follow-up of patients who have received surgery for ovarian carcinoma, since it promises to reliably identify patients who do not require surgical re-exploration, and guide biopsies when they are indicated. PMID- 10682675 TI - IL-6 production in ovarian carcinoma is associated with histiotype and biological characteristics of the tumour and influences local immunity. AB - The presence of interleukin (IL)-6 in peritoneal carcinomatous fluid (PCF) and its effect on immune cells composition in PCF in patients with advanced ovarian carcinoma was studied. In 21 out of 30 ovarian carcinoma patients, PCF IL-6 levels were found to exceed those seen in PCFs of patients with gastrointestinal cancer. IL-6 activity was higher in serous/mucinous than in endometrioid and undifferentiated ovarian carcinoma PCF (P = 0.05). Ovarian carcinoma PCF IL-6 activities were correlated with serum C-reactive protein levels (r = 0.65, P = 0.0000, n = 25). Ovarian carcinoma PCF leucocyte profile differed from that in blood with respect to: (i) lower percentage of NK and CD8+ and (ii) higher percentage of B and CD45RO+, CD14+ and HLA-DR+ cells. The proportions of CD45RO+ in blood were correlated with IL-6 levels in PCF. Corresponding to PCF ovarian carcinoma tumours were stained for the presence of Ki-67 antigen and p53. The highest proportions of Ki-67+ cells and cells showing accumulation of p53 were seen in undifferentiated tumours. A low grade of p53 staining was seen in tumours associated with high IL-6 levels in PCF. It was evident that IL-6 production (i) depended on the histiotype of the tumour, (ii) influenced the local immune system in favour of accumulation of B, and T memory cells, and (iii) was higher in patients lacking p53 accumulation. PMID- 10682676 TI - Mechanistic aspects of the cytotoxic activity of glufosfamide, a new tumour therapeutic agent. AB - Beta-D-glucosyl-ifosfamide mustard (D 19575, glc-IPM, INN = glufosfamide) is a new agent for cancer chemotherapy. Its mode of action, which is only partly understood, was investigated at the DNA level. In the breast carcinoma cell line MCF7 glufosfamide inhibited both the synthesis of DNA and protein in a dose dependent manner, as shown by the decreased incorporation of [3H-methyl] thymidine into DNA and [14C]-methionine into protein of these cells. Treatment of MCF7 cells with 50 microM glufosfamide was sufficient to trigger poly(ADP-ribose) polymerase (PARP) activation, as revealed by immunofluorescence analysis. Both CHO-9 cells, which are O6-methylguanine-DNA methyltransferase (MGMT)-deficient, and an isogenic derivative, which has a high level of MGMT, showed the same cytotoxic response to beta-D-glc-IPM, indicating that the O6 position of guanine is not the critical target for cytotoxicity. By contrast, a sharp decrease in survival of cross-link repair deficient CL-V5 B cells was observed already at concentrations of 0.1 mM beta-D-glc-IPM, whereas the wild-type V79 cells showed a 90% reduction in survival only after treatment with 0.5 mM of this compound. The therapeutically inactive beta-L-enantiomer of glufosfamide also showed genotoxic effects in the same assays but at much higher doses. This was probably due to small amounts of ifosfamide mustard formed under the conditions of incubation. The results indicate that the DNA crosslinks are the most critical cytotoxic lesions induced by beta-D-glc-IPM. PMID- 10682677 TI - Vascular endothelial growth factor expression is independent of hypoxia in human malignant glioma spheroids and tumours. AB - We recently showed that severe hypoxia was not universally present adjacent to necrosis in human glioma xenografts and spheroids established from the M059K, M006, M006X, M006XLo and M010b cell lines. Using these glioma models, we wished to test whether oxygen serves as a regulator of cellular VEGF expression in situ. In situ hybridization (ISH) and immunohistochemistry (IHC) were used to detect vascular endothelial growth factor (VEGF) mRNA and protein expression in sections of glioma xenografts and spheroids in which hypoxic regions and regions with well oxygenated necrosis were identified on contiguous sections by use of the hypoxia specific marker, 3H-misonidazole. Independent validation of the presence of radiobiologically hypoxic cells in M006 xenografts was undertaken using the comet assay. Northern blotting analyses of monolayer cells demonstrated significant up regulation of VEGF mRNA in the M006X line at oxygen concentrations of 6% and below. ISH analysis of VEGF mRNA showed unexpectedly strong staining for VEGF mRNA across the entire viable rim of M006X and M006XLo glioma spheroids. Similarly, in virtually all xenograft tumours of the M059K, M006 and M010b lines, VEGF ISH showed similar staining across all regions of healthy cells up to the border of necrosis. Only in one M006X tumour was there a suggestion of increased VEGF expression in cells adjacent to necrosis. IHC for VEGF showed good concordance with the ISH results. IHC analysis of the VEGF receptor flt-1 showed strong tumour cell staining in M006XLo glioma cells. In human glioma spheroids and xenograft tumours, regions of severe hypoxia do not correspond to areas of up regulated VEGF expression; in fact, VEGF expression is quite uniform. Furthermore, this and our previous study demonstrate that levels of VEGF expression vary among sublines (M006, M006X and M006XLo) derived from a single human glioma specimen. PMID- 10682678 TI - p53-dependent G2 arrest associated with a decrease in cyclins A2 and B1 levels in a human carcinoma cell line. AB - In vivo transfer of wild-type (wt) p53 gene via a recombinant adenovirus has been proposed to induce apoptosis and increase radiosensitivity in several human carcinoma models. In the context of combining p53 gene transfer and irradiation, we investigated the consequences of adenoviral-mediated wtp53 gene transfer on the cell cycle and radiosensitivity of a human head and neck squamous cell carcinoma line (SCC97) with a p53 mutated phenotype. We showed that ectopic expression of wtp53 in SCC97 cells resulted in a prolonged G1 arrest, associated with an increased expression of the cyclin-dependent kinase inhibitor WAF1/p21 target gene. A transient arrest in G2 but not in G1 was observed after irradiation. This G2 arrest was permanent when exponentially growing cells were transduced by Ad5CMV-p53 (RPR/INGN201) immediately after irradiation with 5 or 10 Gy. Moreover, levels of cyclins A2 and B1, which are known to regulate the G2/M transition, dramatically decreased as cells arrived in G2, whereas maximal levels of expression were observed in the absence of wtp53. In conclusion, adenoviral mediated transfer of wtp53 in irradiated SCC97 cells, which are mutated for p53, appeared to increase WAF1/p21 expression and decrease levels of the mitotic cyclins A2 and B1. These observations suggest that the G2 arrest resulted from a p53-dependent premature inactivation of the mitosis promoting factor. PMID- 10682679 TI - NADPH:cytochrome c (P450) reductase activates tirapazamine (SR4233) to restore hypoxic and oxic cytotoxicity in an aerobic resistant derivative of the A549 lung cancer cell line. AB - Tirapazamine (TPZ, SR4233, WIN 59075) is a bioreductive drug that is activated in regions of low oxygen tension to a cytotoxic radical intermediate. This labile metabolite shows high selective toxicity towards hypoxic cells, such as those found in solid tumours. Under aerobic conditions, redox cycling occurs with subsequent generation of superoxide radicals, which are also cytotoxic. NADPH:cytochrome c (P450) reductase (P450R) is a one-electron reducing enzyme that efficiently activates TPZ. Recently a derivative of the A549 non-small cell lung cancer cell line (A549c50) was generated that showed substantially reduced P450R activity compared to its parental line (Elwell et al (1997) Biochem Pharmacol 54: 249-257). Here, it is demonstrated that the A549c50 cells are markedly more resistant to TPZ under both aerobic and hypoxic conditions. In addition, these cells have a dramatically impaired ability to metabolize TPZ to its two-electron reduction product, SR4317, under hypoxic conditions when compared to wild-type cells. P450R activity in the A549c50 cells was reintroduced to similar levels as that seen in the parental A549 cells by transfection of the full-length cDNA for human P450R. These P450R over-expressing cells exhibit restored sensitivity to TPZ under both aerobic and hypoxic conditions, comparable to that found in the original parental A549 cells. Further, the ability of the transfected cells to metabolize TPZ to SR4317 under hypoxic conditions is also shown to be restored. This provides further evidence that P450R can play an important role in the activation, metabolism and toxicity of this lead bioreductive drug. PMID- 10682681 TI - ERBB-2 overexpression confers PI 3' kinase-dependent invasion capacity on human mammary epithelial cells. AB - Amplification and overexpression of ERBB-2 in human breast cancer is thought to play a significant role in the progression of the disease; however, its precise role in the aetiology of altered phenotypes associated with human breast cancer is unknown. We have previously shown that exogenous overexpression of ERBB-2 conferred growth factor independence on human mammary epithelial cells. In this study, we show that ERBB-2 overexpression also causes the cells to acquire other characteristics exhibited by human breast cancer cells, such as anchorage independent growth and invasion capabilities. ERBB-2-induced invasion is dependent on fibronectin and correlates with the down-regulation of cell surface alpha4 integrin. In addition ERBB-2 co-immunoprecipitates with focal adhesion kinase (FAK) in these cells. We have also shown, by use of exogenously expressed PTEN and by treatment with the PI3'-kinase inhibitor LY294002, that ERBB-2 induced invasion is dependent on the PI3'-kinase pathway; however, PTEN does not dephosphorylate FAK in these cells. PMID- 10682680 TI - Collagenolytic and gelatinolytic matrix metalloproteinases and their inhibitors in basal cell carcinoma of skin: comparison with normal skin. AB - Tissue from 54 histologically-identified basal cell carcinomas of the skin was obtained at surgery and assayed using a combination of functional and immunochemical procedures for matrix metalloproteinases (MMPs) with collagenolytic activity and for MMPs with gelatinolytic activity. Collagenolytic enzymes included MMP-1 (interstitial collagenase), MMP-8 (neutrophil collagenase) and MMP-13 (collagenase-3). Gelatinolytic enzymes included MMP-2 (72-kDa gelatinase A/type IV collagenase) and MMP-9 (92-kDa gelatinase B/type IV collagenase). Inhibitors of MMP activity including tissue inhibitor of metalloproteinases-1 and -2 (TIMP-1 and TIMP-2) were also assessed. All three collagenases and both gelatinases were detected immunochemically. MMP-1 appeared to be responsible for most of the functional collagenolytic activity while gelatinolytic activity reflected both MMP-2 and MMP-9. MMP inhibitor activity was also present, and appeared, based on immunochemical procedures, to reflect the presence of TIMP-1 but not TIMP-2. As a group, tumours identified as having aggressive-growth histologic patterns were not distinguishable from basal cell carcinomas with less aggressive-growth histologic patterns. In normal skin, the same MMPs were detected by immunochemical means. However, only low to undetectable levels of collagenolytic and gelatinolytic activities were present. In contrast, MMP inhibitor activity was comparable to that seen in tumour tissue. In previous studies we have shown that exposure of normal skin to epidermal growth factor in organ culture induces MMP up-regulation and activation. This treatment concomitantly induces stromal invasion by the epithelium (Varani et al (1995) Am J Pathol 146: 210-217; Zeigler et al (1996b) Invasion Metastasis 16: 11 18). Taken together with these previous data, the present findings allow us to conclude that the same profile of MMP/MMP inhibitors that is associated with stromal invasion in the organ culture model is expressed endogenously in basal cell carcinomas of skin. PMID- 10682682 TI - The role of p16-cyclin d/CDK-pRb pathway in the tumorigenesis of endometrioid type endometrial carcinoma. AB - We analysed p16 gene alteration and p16, cyclin-dependent kinase 4 (CDK4), CDK6, cyclin D1, cyclin D2, cyclin D3 and retinoblastoma protein (pRb) expression in ten normal endometriums (PE), 18 endometrial hyperplasias (EH) and 35 endometrial cancers (EC). Two of ten PE (20%), nine of 18 EH (50.0%) and 29 of 35 EC (82.9%) exhibited p16 nuclear staining. p16 expression was significantly higher in EC than EH (P = 0.0119). In the six p16 (-) EC, one was considered to have reduced gene dosage consistent with possible homozygous deletion of the CDKN2 gene and three had methylation in 5'CpG island in the promoter region of the p16 gene, whereas none showed such reduced gene dosage and four had methylation in the nine p16 (-) EH. Strong CDK4 staining was observed in 12 of 35 EC (34.3%) and one of 18 EH (5.6%). The strong expression of CDK4 was higher in EC than in EH (P = 0.0399). The expression of CDK4 was higher in EH than PE (P = 0.0054). The abnormalities of p16-cyclin D/CDK-pRb pathway were detected in 18 of 35 EC (51.4%). In conclusion, the expression of p16 and CDK4 may be an early event in the neoplastic transformation of endometrial cancer. PMID- 10682683 TI - Interaction of the PA2G4 (EBP1) protein with ErbB-3 and regulation of this binding by heregulin. AB - The processes by which ErbB-3, an inactive tyrosine kinase, exerts its biological effects are poorly understood. Using the yeast two-hybrid system, we have isolated an ErbB-3 binding protein (Ebp1) that interacts with the juxtamembrane domain of ErbB-3. This protein is identical to that predicted to be encoded for by the human PA2G4 gene. Ebp1 is the human homologue of a previously identified cell cycle-regulated mouse protein p38-2G4. Two transcripts of ebp1 mRNA (1.7 and 2.2 kb) were detected in several normal human organs. The interaction of Ebp1 with ErbB-3 was examined in vitro and in vivo. The first 15 amino acids of the juxtamembrane domain of ErbB-3 were essential for Ebp1 binding in vitro. Treatment of AU565 cells with the ErbB-3 ligand heregulin resulted in dissociation of Ebp1 from ErbB-3. Ebp1 translocated from the cytoplasm into the nucleus following heregulin stimulation. These findings suggest that Ebp1 may be a downstream member of an ErbB-3-regulated signal transduction pathway. PMID- 10682685 TI - Detection of antibodies to human herpesvirus 8 in Italian children: evidence for horizontal transmission. AB - Human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma associated herpesvirus (KSHV), has been shown to be the causative agent for Kaposi's sarcoma (KS) and to be more prevalent in populations or risk groups at increased risk for KS. HHV-8 infection is rare in children from the US and the UK, but has been reported in African children. In this study we examine HHV-8 infection in children from Italy, a country with an elevated prevalence of HHV-8 in adults and high socio-economic conditions. PMID- 10682684 TI - Lack of evidence for MHC-unrestricted (atypical) recognition of mucin by mucinous pancreatic tumour-reactive T-cells. AB - Cytotoxic T-cells generated against heterologous, mucinous pancreatic tumour cells were shown to recognize mucin in a major histocombatibility complex (MHC) unrestricted fashion. In contrast, the present study demonstrates a typical allogeneic response of heterologous cytotoxic T-cells established against mucin expressing pancreatic tumour cells. Heterologous cytotoxic T cells lysed targets that were used as stimulators and other targets that shared human leucocyte antigen (HLA) with the stimulator. These cytotoxic T-cells lysed mucin-expressing stimulator cells but not autologous tumour cells in spite of expressing mucin on their surface. Likewise, tumour-infiltrating CD4+ T-cells proliferated against its own tumour cell target, while such T-cells did not respond to heterologous, mucin-expressing pancreatic tumour cells. Culturing heterologous tumour-specific cytotoxic T-cells with purified pancreatic tumour cell-mucin rendered them unresponsive to their target cells. Furthermore, purified mucin did not produce a mucin-specific response in mucinous pancreatic tumour patients' primary T-cells even in the presence of antigen-presenting cells. Our study finds no evidence for MHC-unrestricted recognition of mucin by pancreatic cancer patients' T-cells. PMID- 10682686 TI - Evidence of a founder BRCA1 mutation in Scotland. AB - BRCA1 mutations have been identified in breast and ovarian cancer families from diverse ethnic backgrounds. We studied 17 different families with the BRCA1 2800delAA mutation; seven were ascertained in Scotland (Dundee, Edinburgh, Glasgow, St Andrews), five in Canada (Toronto, Victoria) and five in the United States (Chicago, Philadelphia, Seattle). Overall there was a clear preponderance of Scottish ancestry. Genotype analysis performed on key members from 17 families was consistent with a common haplotype, strongly suggesting a single ancestral origin. A possible link was established between two families by tracing their genealogies through the records of the Registrar General for Scotland. This is the first example of a BRCA1 mutation likely to be derived from a common founder in Scotland. Further studies will be necessary to estimate more accurately the population frequency of the BRCA1 2800delAA mutation among unselected cases of breast and ovarian cancer in Scotland and the UK. PMID- 10682687 TI - Reducing DCO registrations through electronic matching of cancer registry data and routine hospital data. AB - The Thames Cancer Registry (TCR) has registered a high proportion of tumours from death certificate information only (DCO) registrations. This paper describes the results of a study set up to establish whether this proportion could be reduced by linking cancer registrations with routine hospital data from the Hospital Episodes Statistics (HES) data set using computerized matching. A total of 67752 registrations were identified from the TCR. Matches were found in the HES data set for 66%. The proportion of cases retrieved for each tumour site was: 72% for colorectal cancer; 62% for cancer of the lung, trachea or bronchus; and 65% for female breast cancer. For all three tumour sites the proportion of matches found for patients registered from hospital case notes was higher than the proportion found for patients registered as DCOs (P < 0.0001 for all three tumour sites). Among matched DCO cases, 58% had at least one procedure recorded. DCO rates might be reduced by as much as 43% (from 17% of total registrations to less than 10%) for the three most common cancers if the method of electronic matching outlined here was used. Younger age groups, prognosis of tumour site and residence in North Thames region were all positively associated with successful matching (P < 0.0001 in all three cases). Many matched DCO cases were found to have had more than one admission for cancer. Among ordinary in-patient admissions, admissions to patients ratios of 1.5, 1.4 and 1.9 were found for colorectal, lung and breast cancers respectively. Of 5190 matched DCOs a procedure was recorded for 3013 (58%). HES data offer a useful aid to follow-up of case notes on patients identified to the registry by death certificates. Doubts about the completeness and accuracy of HES data mean case notes must remain the 'gold standard'. PMID- 10682689 TI - Breast cancer among former college athletes compared to non-athletes: a 15-year follow-up. AB - A growing body of evidence indicates that physical activity is protective against breast cancer. In 1996-97, we conducted a 15-year follow-up of 5398 college alumnae comprised of former college athletes with their non-athletic classmates. Participants completed a detailed mailed questionnaire on their health history from 1981-82 to the present. Excluding women who had died and non-deliverable questionnaires, 84.7% (n = 3940) of the participants in our earlier study responded to the questionnaire; the response rate for former athletes was 86.6% (n = 1945), for non-athletes, 83.0% (n = 1995). Results confirmed our earlier findings. Based on self-reports, former college athletes had a significantly lower risk of breast cancer than the non-athletes. The OR for the 15-year incidence of breast cancer is 0.605 with 95% confidence interval (CI) (0.438 0.835); the 15-year incident breast cancers were 64 among the athletes and 111 among the non-athletes. Among women under 45 the protective effect of physical activity on the risk of breast cancer is considerably greater; odds ratio (OR) = 0.164, 95% CI (0.042-0.636). Athletic activity during the college and pre-college years is protective against breast cancer throughout the life span, and more markedly among women under 45. These results confirm our earlier findings and the findings of other investigators. PMID- 10682688 TI - Sexual behaviour, STDs and risks for prostate cancer. AB - A population-based case-control study was carried out among 981 men (479 black, 502 white) with pathologically confirmed prostate cancer and 1315 controls (594 black, 721 white). In-person interviews elicited information on sexual behaviour and other potential risk factors for prostate cancer. Blood was drawn for serologic studies in a subset of the cases (n = 276) and controls (n = 295). Prostate cancer risk was increased among men who reported a history of gonorrhoea or syphilis (odds ratio (OR) = 1.6; 95% confidence internal (CI) 1.2-2.1) or showed serological evidence of syphilis (MHA-TP) (OR = 1.8; 95% CI 1.0-3.5). Patterns of risk for gonorrhoea and syphilis were similar for blacks (OR = 1.7; 95% CI 1.2-2.2) and whites (OR = 1.6; 95% CI 0.8-3.2). Risks increased with increasing occurrences of gonorrhoea, rising to OR = 3.3 (95% CI 1.4-7.8) among subjects with three or more events (Ptrend = 0.0005). Frequent sexual encounters with prostitutes and failure to use condoms were also associated with increased risk. Syphilis, gonorrhoea, sex with prostitutes and unprotected sexual intercourse may be indicators of contact with a sexually transmissible factor that increases the risk of prostate cancer. PMID- 10682690 TI - Adding free to total prostate-specific antigen levels in trials of prostate cancer screening. AB - We used a nested case-control design on data from men in four prospective studies (from the UK, Maryland in the USA, and two from Finland) with available stored serum samples to determine whether there was an advantage in measuring both free prostate-specific antigen (PSA) and total PSA as a potential screening test for prostate cancer. Of these men, 247 were verified through national vital statistics offices as having died of prostate cancer, or having developed the disease, and 953 men who did not develop prostate cancer (controls) were selected, matched to cases for age, study centre and sample storage duration. Fixing the false-positive rate at 1%, the prostate cancer detection rate (sensitivity) over the 3 years following serum collection (based on 14 cancers) increased from an estimated 95% using total PSA to 97% using free and bound PSA (that is, bound to alpha-antichymotrypsin which together with the free form is total PSA). Over a 6-year period (based on 41 cancers) a similar difference occurred (52% and 56% detection rates respectively). We conclude that there is no material advantage in adding free to total PSA in prostate cancer screening trials. PMID- 10682691 TI - Fertility in Norwegian testicular cancer patients. AB - The intention was to explore the relationship between fertility and testicular cancer, including the possibly treatment-induced changes over time in the post diagnostic fertility. Data are from the Norwegian Cancer Registry, The Norwegian Population Register and the Population Censuses. By estimating Poisson regression models, birth rates among testicular cancer patients were compared with those of other men who had the same age, parity and duration since previous birth. Poisson regression models were also estimated to check whether men's parity has an effect on the cancer incidence. Fertility rates among testicular cancer patients born after 1935 and treated before 1991 decreased by roughly 30% when compared with the normal population. The introduction of cisplatin chemotherapy and of nerve sparing RPLND in the 1980s seems to have enabled more patients with non-seminoma to father a child after treatment, or at least shortened the time to conception. Moreover, the risk of being diagnosed with seminoma is reduced with increasing parity. This suggests that the relatively low fertility after diagnosis may be partly due to the continuing inherent influence of a sub- or infecundity that also had a bearing on the development of the disease. PMID- 10682692 TI - Delay in presentation of symptomatic referrals to a breast clinic: patient and system factors. AB - We attempted to identify factors associated with delay in presentation and assessment of women with breast symptoms who attended a London breast clinic. A total of 692 consecutive symptomatic referrals, aged 40-75 years, were studied. Patient delay, assessed prior to diagnosis, was defined as time elapsing between symptom discovery and first presentation to a medical provider. This was studied in relation to: reasons for delaying, beliefs and attitudes, socio-demographic and clinical variables, psychiatric morbidity and subsequent diagnosis. Thirty five per cent of the cohort delayed presentation 4 weeks or more (median 13 days). The most common reason given was that they thought their symptom was not serious (odds ratio (OR) = 5.32, 95% confidence interval (CI) 3.6-8.0). Others thought their symptom would go away (OR = 3.73, 95% CI 2.2-6.4) or delayed because they were scared (OR = 4.61, 95% CI 2.1-10.0). Delay was associated with psychiatric morbidity but not age. Patients who turned out to have cancer tended to delay less (median 7 days) but not significantly. Median system delay--time between first medical consultation and first clinic visit--was 18 days. Patients who thought they had cancer and those so diagnosed were seen more promptly (median 14 days). Most factors, including socio-economic status and ethnicity were non-contributory. Beliefs about breast symptoms and their attribution are the most important factors determining when women present. Health education messages should aim to convince symptomatic women that their condition requires urgent evaluation, without engendering fear in them. PMID- 10682693 TI - Retrospective study of the relationships between age, inflammation and the isolation of bacteria from the lower respiratory tract of thoroughbred horses. AB - A total of 1235 tracheal aspirates taken from 724 thoroughbreds in race training, aged from two to 10 years, were examined cytologically and bacteriologically. An inflammation scoring system on a scale of 0 to 9 was devised to allow the severity of lower airway disease to be assessed from the cytological results. The inflammation scores were closely related to the isolation of bacteria (P<0.001), and the most common bacterial isolates were Streptococcus zooepidemicus, Streptococcus pneumoniae and Pasteurella/Actinobacillus-like species. Lower airway disease was less common in older horses (P = 0.031), and the groups at highest risk were the two- and four-year-olds. Lower airway inflammation was more common in the four-year-olds at National Hunt yards than in the four-year-olds at flat racing yards (P = 0.040, odds ratio = 3.80). PMID- 10682694 TI - Survey of retinal haemorrhages in neonatal thoroughbred foals. AB - Twenty-seven of 167 neonatal thoroughbred foals (16 per cent) were found to have retinal haemorrhages after a careful examination of the entire fundus. Experience in differentiating haemorrhages from other lesions, and the selection of foals from normal populations, were considered to have an important effect on their apparent incidence. Bilateral haemorrhages were more common and there was a significantly higher incidence in female foals. The numbers of haemorrhages ranged between one and 20; 36 per cent of eyes with retinal haemorrhages had the small punctate type and 56 per cent had the splash-like form. There was no change from one type of haemorrhage to another, and the patterns of resolution were similar. The haemorrhages were in the tapetal fundus, except two that were recorded in the optic disc, and they resolved within 10 days. No short- or long term ocular or neurological effects of the retinal haemorrhages were detected, and they were not related to the incidence of abnormal foal behaviour. PMID- 10682695 TI - Transfer of chlortetracycline from contaminated feedingstuff to cows' milk. AB - Three groups of four Friesian cows in mid-lactation were fed a compound feedingstuff contaminated with 2, 10 or 300 mg chlortetracycline/kg for 21 days, and were then fed an uncontaminated diet for seven days. A fourth group of four cows was fed an uncontaminated diet throughout the study. Daily pooled milk samples from each cow were analysed by high performance liquid chromatography (HPLC) with a detection limit of 50 microg chlortetracycline/litre. Chlortetracycline was detected in only two milk samples taken from one of the animals fed feed containing 300 mg 300 mg chlortetracycline/kg, and both contained less than the maximum residue limit (MRL) specified by the European Union (100 microg/litre). All the milk samples were also analysed by the Delvotest SP microbiological assay, which has a detection limit of 300 microg chlortetracycline/litre. During the treatment period, this method gave four presumptive false-positive results, because they were not confirmed by HPLC. Selected daily pooled samples from each treatment group were also analysed by the semi-quantitative Charm II radioreceptor assay with a detection limit of 10 microg chlortetracycline/litre. Immunoreactive chlortetracycline was detected only in the animals fed feed containing 300 mg chlortetracycline/kg and several of the results exceeded the EU MRL during the treatment period. No significant treatment effects on animal performance were observed. However, there was a trend towards a higher milk fat concentration (P<0.09) and a lower milk protein concentration (P<0.07) with increasing concentration of chlortetracycline in the diet. PMID- 10682696 TI - African horse sickness in Senegal: serotype identification and nucleotide sequence determination of segment S10 by RT-PCR. PMID- 10682697 TI - Transport stress in cattle as reflected by an increase in faecal cortisol metabolite concentrations. PMID- 10682698 TI - Student debt and socioeconomic mix. PMID- 10682699 TI - Clomipramine and behavioural therapy in the treatment of separation-related problems in dogs. PMID- 10682700 TI - Inositol trisphosphate and cyclic adenosine diphosphate-ribose increase quantal transmitter release at frog motor nerve terminals: possible involvement of smooth endoplasmic reticulum. AB - The release of chemical transmitter from nerve terminals is critically dependent on a transient increase in intracellular Ca2+. The increase in Ca2+ may be due to influx of Ca2+ from the extracellular fluid or release of Ca2+ from intracellular stores such as mitochondria. Whether Ca2+ utilized in transmitter release is liberated from organelles other than mitochondria is uncertain. Smooth endoplasmic reticulum is known to release Ca2+, e.g., on activation by inositol trisphosphate or cyclic adenosine diphosphate-ribose, so the possibility exists that Ca2+ from this source may be involved in the events leading to exocytosis. We examined this hypothesis by testing whether inositol trisphosphate and cyclic adenosine diphosphate-ribose modified transmitter release. We used liposomes to deliver these agents into the cytoplasmic compartment and binomial analysis to determine their effects on the quantal components of transmitter release. Administration of inositol trisphosphate (10(-4)M) caused a rapid, 25% increase in the number of quanta released. This was due to an increase in the number of functional release sites, as the other quantal parameters were unaffected. The effect was reversed with 40 min of wash. Virtually identical results were obtained with cyclic adenosine diphosphate-ribose (10(-4)M). Inositol trisphosphate caused a 10% increase in quantal size, whereas cyclic adenosine diphosphate-ribose had no effect. The results suggest that quantal transmitter release can be increased by Ca2+ released from smooth endoplasmic reticulum upon stimulation by inositol trisphosphate or cyclic adenosine diphosphate-ribose. This may involve priming of synaptic vesicles at the release sites or mobilization of vesicles to the active zone. Inositol trisphosphate may have an additional action to increase the content of transmitter within the vesicles. These findings raise the possibility of a role of endogenous inositol phosphate and smooth endoplasmic reticulum in the regulation of cytoplasmic Ca2+ and transmitter release. PMID- 10682701 TI - Cortical cholinergic inputs mediating arousal, attentional processing and dreaming: differential afferent regulation of the basal forebrain by telencephalic and brainstem afferents. AB - Basal forebrain corticopetal neurons participate in the mediation of arousal, specific attentional functions and rapid eye movement sleep-associated dreaming. Recent studies on the afferent regulation of basal forebrain neurons by telencephalic and brainstem inputs have provided the basis for hypotheses which, collectively, propose that the involvement of basal forebrain corticopetal projections in arousal, attention and dreaming can be dissociated on the basis of their regulation via major afferent projections. While the processing underlying sustained, selective and divided attention performance depends on the integrity of the telencephalic afferent regulation of basal forebrain corticopetal neurons, arousal-induced attentional processing (i.e. stimulus detection, selection and processing as a result of a novel, highly salient, aversive or incentive stimuli) is mediated via the ability of brainstem ascending noradrenergic projections to the basal forebrain to activate or "recruit" these telencephalic afferent circuits of the basal forebrain. In rapid eye movement sleep, both the basal forebrain and thalamic cortiocopetal projections are stimulated by cholinergic afferents originating mainly from the pedunculopontine and laterodorsal tegmenta in the brainstem. Rapid eye movement sleep-associated dreaming is described as a form of hyperattentional processing, mediated by increased activity of cortical cholinergic inputs and their cortical interactions with activated thalamic efferents. In this context, long-standing speculations about the similarities between dreaming and psychotic cognition are substantiated by describing the role of an over(re)active cortical cholinergic input system in either condition. Finally, while determination of the afferent regulation of basal forebrain corticopetal neurons in different behavioral/cognitive states assists in defining the general cognitive functions of cortical acetylcholine, this research requires a specification of the precise anatomical organization of basal forebrain afferents and their interactions in the basal forebrain. Furthermore, the present hypotheses remain incomplete because of the paucity of data concerning the regulation and role of basal forebrain non-cholinergic, particularly GABAergic, efferents. PMID- 10682702 TI - Cortical and subcortical influences on the nucleus of the optic tract of the opossum. AB - In the present work we propose a new phylogenetic hypothesis for the role played by cortical and subcortical afferents to the nucleus of the optical tract, the main visual relay station of the horizontal optokinetic reflex in mammals. The hypothesis is supported by anatomical and physiological data obtained in the South American opossum (Didelphis aurita) using the following experimental approaches: (i) single-unit recordings in the nucleus of the optic tract and simultaneous electrical stimulation of the contralateral nucleus of the optic tract; (ii) single-unit recordings in the nucleus of the optic tract and simultaneous electrical stimulation of the ipsilateral striate cortex; (iii) injection of cholera toxin subunit B into the striate cortex and subsequent immunohistochemical reaction to reveal the presence of the marker in the thalamus and mesencephalon; and (iv) single-unit recordings in the nucleus of the optic tract both before and after ablation of the ipsilateral visual cortex. The main results are: (i) there is a strong inhibitory reciprocal effect upon the nucleus of the optic tract following stimulation of its contralateral counterpart; (ii) electrophysiological and anatomical data imply that the visual cortex does not project directly to the nucleus of the optic tract. Rather, cortical terminals seem to target the nearby anterior and posterior pretectal nuclei and orthodromic latencies in the nucleus of the optic tract following stimulation of the visual cortex were twice as large as in the superior colicullus; and (iii) ablation of the entire visual cortex did not have any effect upon binocularity of cells in the nucleus of the optic tract. These results strengthen the model proposed here for the role of the interactions between the nuclei of the optic tract under optokinetic stimulation. The hypothesis in the present work is that the cortical influences upon the nucleus of the optical tract, in addition to the subcortical ones, appeared only recently in phylogenesis. In more primitive mammals, such as the opossum, subcortical interactions are thought to play a relatively important role. With the emergence of retinal specializations, such as the fovea, one might suppose that there followed the appearance of new ocular movements, such as the smooth pursuit and certain types of saccades, that came to join the pre-existent optokinetic reflex. PMID- 10682703 TI - Subthreshold inward membrane currents in guinea-pig frontal cortex neurons. AB - Current-clamp and single-electrode voltage-clamp recordings were used to study the inward currents activated in the subthreshold membrane potential range of cortical pyramidal neurons. The experiments were done on slices from guinea-pig frontal cortex and all recordings were obtained at a distance of 600-900 microm from the pial surface. In current-clamp recordings and from membrane potentials hyperpolarized to about -70 mV, the depolarization leading to spike firing was partially blocked by 1 microM tetrodotoxin, but not by calcium-free extracellular solution. The calcium-free solution only affected this depolarization when the membrane potential was held at a level more negative than -75 mV. Under voltage clamp, an inward current was recorded between the resting membrane potential and the level of spike firing. This current was activated at about -60 mV and part of it was blocked by 1 microM tetrodotoxin; the remaining current was blocked by calcium-free extracellular solution. In five neurons both components were recorded and isolated in the same cell. The tetrodotoxin-sensitive component activated at close to -60 mV, was similar to the persistent sodium current (I(Na p)). The Ca2+-sensitive component activated at close to -60 or -65 mV, was less voltage-dependent than I(Na-p). This component was similar to the low threshold calcium current (I(T)). These results suggest that the subthreshold depolarization which led to spike firing was dependent on I(Na-p) and I(T), I(Na p) being the most important factor up to resting membrane potentials of -70 or 75 mV. A physiological role of this finding is revealed by the action of dopamine, which (at 10 microM) prevented the firing of action potentials from -60 mV, but not from -80 mV due to the inhibition of I(Na-p) and the lack of effect on I(T). PMID- 10682704 TI - Subcellular localization of glutamate-stimulated intracellular magnesium concentration changes in cultured rat forebrain neurons using confocal microscopy. AB - Glutamate can stimulate increases in intracellular magnesium concentration ([Mg2+]i) and induce neurotoxicity, both independent of Ca2+ changes. Although Mg2+ is essential within the cell, very little is known about how it is regulated, especially in neurons. Therefore we used the fluorescent indicator, magindo-1 and confocal microscopy to examine possible intracellular pools of Mg2+ in cultured neurons that can be dynamically regulated by glutamate. The magindo-1 fluorescence signal was present throughout the cell body and extends into the neuronal processes. The magindo-1 405 nm/490 nm ratio signal was similar in the cytoplasm and nucleus, suggesting that resting [Mg2+]i is uniform across the neuron. The addition of 100 microM glutamate/10 microM glycine in an extracellular Ca2+- and Na+-free buffer stimulated an increase in [Mg2+]i in both the nuclear and cytoplasmic regions of similar magnitude and duration. This glutamate exposure also stimulated a [Mg2+]i increase in neuronal processes which was inhibited by the N-methyl-D-aspartate receptor antagonist, MK-801 (10 microM). The glutamate-stimulated [Mg2+]i increase in both the cell body and neuronal processes was dependent on the extracellular Mg2+ concentration. These findings suggest glutamate-stimulated [Mg2+]i changes may not only impact cytoplasmic processes, but also directly trigger nuclear events involved, for example, in neuronal injury. PMID- 10682705 TI - Gerbil angiotensin II AT1 receptors are highly expressed in the hippocampus and cerebral cortex during postnatal development. AB - Increasing evidence suggests that Angiotensin II, classically known from its many effects regulating salt and water homeostasis, is also involved in brain development and cognitive functions through activation of AT1 Angiotensin II receptors. The recently cloned gerbil AT1 receptor is expressed in brain areas controlling hydro-mineral homeostasis, and particularly highly expressed in limbic areas such as the hippocampal formation. We quantified the gerbil AT1 receptor messenger RNA expression and receptor binding by quantitative in situ hybridization and receptor autoradiography, respectively, in the hippocampal formation and cerebral cortex of gerbils during postnatal development. The receptor messenger RNA and binding were present from birth and showed a gradual and sustained increase through postnatal maturation in the CA1 and CA2 regions of the hippocampus and in the dentate gyrus. Conversely, in the CA3 region, no binding was detected while receptor messenger RNA peaked at 15 days after birth and disappeared in the adult. The highest receptor messenger RNA expression and binding were found in the septomedial portions of the CA1 region and at septal levels of the CA2 region. We detected the highest receptor messenger RNA expression at postnatal day one in the frontolateral pole of the cerebral hemispheres. In these areas, and in the frontoparietal and insular cortex, receptor messenger RNA dramatically decreased during postnatal life. Similarly, we found receptor messenger RNA expression in the cingulate, retrosplenial, perirhinal and infralimbic cortex with higher values during the first two weeks of development and decreased expression in the adult. However, receptor binding in the cerebral cortex, did not decrease during postnatal life. The differential profile of receptor messenger RNA expression and binding in the gerbil cortex and hippocampus during postnatal maturation suggest a role for AT1 receptors in the development and function of the corticohippocampal system. PMID- 10682706 TI - Endothelin b receptor deficiency is associated with an increased rate of neuronal apoptosis in the dentate gyrus. AB - The dentate gyrus retains neuronal proliferative potential throughout life. Using immature endothelin B receptor-deficient (sl/sl) rats, a rabbit model of pneumococcal meningitis and autopsy brains from humans who died from pneumococcal meningitis, we explored the role of endothelin B receptors in physiological and pathological neuronal apoptosis in the dentate gyrus. At postnatal days 3-4, the rate of apoptosis in the dentate gyrus was high in all rats, declining to low levels in wild-type rats (+/+) on days 14 and 22, but remaining high in both homozygous (sl/sl) and heterozygous (sl/+) endothelin B receptor-deficient rats. Increased apoptosis was not significantly compensated for by neuronal proliferation. Hippocampal neuronal cultures also exhibited genotype-dependent apoptosis with the highest rate in neurons from homozygous endothelin B receptor deficient (sl/sl) rats. In rabbit and human pneumococcal meningitis, increased apoptosis in the dentate gyrus was associated with loss of neuronal endothelin B receptor immunoreactivity. In conclusion, endothelin B receptors appear to act as neuronal survival factors in the dentate gyrus in rodents and man, both during postnatal development and under pathological conditions. PMID- 10682707 TI - Expression of brain-derived neurotrophic factor, neurotrophin-3 and their receptor messenger RNAs in monkey rhinal cortex. AB - The primate rhinal cortex, consisting of areas 36 and 35 of the perirhinal cortex and the entorhinal cortex (area 28), plays a crucial role in perception and memory. We investigated the expression of messenger RNAs for brain-derived neurotrophic factor and neurotrophin-3, as well as those for their respective tyrosine kinase receptors, TrkB and TrkC, in the monkey rhinal cortex. Results from in situ hybridization revealed that each of these messenger RNAs was expressed in neurons with distinct laminar and areal patterns of distribution. Brain-derived neurotrophic factor messenger RNA was principally detected in layers V/ VI of area 36, and layers II/III and V of the entorhinal cortex. Some of the messenger RNA-positive cells in the deep layers of the rhinal cortex were confirmed to exhibit a pyramidal cell-like morphology. Neurotrophin-3 messenger RNA expression was confined to layers II/III of the entorhinal cortex. In contrast, trkB and trkC messenger RNAs were expressed rather homogeneously and abundantly throughout the rhinal cortex. The laminar and cellular distributions of brain-derived neurotrophic factor and neurotrophin-3 messenger RNAs indicate the predominant expression of these neurotrophins in projection neurons. These results suggest that brain-derived neurotrophic factor and neurotrophin-3 regulate neuronal connectivities of forward and backward projections from the rhinal cortex and contribute to functional reorganization underlying the formation and maintenance of long-term memory in primates. PMID- 10682708 TI - Glial cell line-derived neurotrophic factor receptor alpha1 availability regulates glial cell line-derived neurotrophic factor signaling: evidence from mice carrying one or two mutated alleles. AB - Glial cell line-derived neurotrophic factor receptor alpha1 (GFRalpha1, also known as GDNFR-alpha) is a glycolipid-anchored membrane protein of the GFRalpha family, which binds glial cell line-derived neurotrophic factor [Jing S. et al. (1996) Cell 85, 1113-1124; Treanor J. J. et al. (1996) Nature 382, 80-83], a survival factor for several populations of central and peripheral neurons, including midbrain dopamine neurons [Lin L. F. et al. (1993) Science 260, 1130 1132], and mediates its ligand-induced cell response via a tyrosine kinase receptor called Ret [Takahashi M. et al. (1988) Oncogene 3, 571-578; Takahashi M. and Cooper G. M. (1987) Molec. Cell Biol. 7, 1378-1385]. In this paper, we show that mice with a null mutation of the GFRalpha1 gene manifest epithelial mesenchymal interaction deficits in kidney and severe disturbances of intestinal tract development similar to those seen with glial cell line-derived neurotrophic factor or Ret null mutations. There is a marked renal dysgenesis or agenesis and the intrinsic enteric nervous system fails completely to develop. We also show that newborn GFRalpha1-deficient mice display no or minimal changes in dorsal root and sympathetic ganglia. This is in contrast to the deficits reported in these neuronal populations in glial cell line-derived neurotrophic factor and Ret null mutations. Mesencephalic dopaminergic neurons in the substantia nigra and ventral tegmental area appear intact at the time of birth of the mutated mice. Mice homozygous for the GFRalpha1 null mutation die within 24 h of birth because of uremia. Heterozygous animals, however, live to adulthood. There is a significantly reduced neuroprotective effect of glial cell line-derived neurotrophic factor in such heterozygous animals, compared with wild-type littermates, after cerebral ischemia. Taken together with previous data on glial cell line-derived neurotrophic factor and Ret, our results strongly suggest that GFRalpha1 is the essential GFRalpha receptor for signaling in the glial cell line derived neurotrophic factor-Ret pathway in the kidney and enteric nervous system development, and that GFRalpha2 or GFRalpha3 cannot substitute for the absence of GFRalpha1. Moreover, neuroprotective actions of exogenous glial cell line-derived neurotrophic factor also require full GFRalpha1 receptor expression. PMID- 10682709 TI - Early direct and transneuronal effects in mice with targeted expression of a toxin gene to D1 dopamine receptor neurons. AB - The neurochemical profile was examined at postnatal day 3-4 in mutant mice generated by in vivo Cre mediated activation of an attenuated diphtheria toxin gene inserted into the D1 dopamine receptor gene locus. An earlier study of this model had shown that D1 dopamine receptor, substance P and dynorphin were not expressed in the striatum. Quantitative in situ hybridization analysis showed an increase in D2 dopamine receptor and enkephalin messenger RNA expression. The nigrostriatal pathway in the mutant pups was intact with a normal number of dopaminergic neurons in the substantia nigra and the ventral tegmental area in addition to a normal pattern of striatal dopamine transporter and tyrosine hydroxylase immunoreactivity. Quantitative analysis of striatal dopamine transporter density using [3H]mazindol showed a reduction of 26% suggesting a degree of transneuronal down-regulation. There was also a 49% reduction of striatal GABA receptor binding and a 36% reduction of striatal muscarinic receptor binding in mutant pups. The number of healthy striatal neuropeptide Y containing interneurons was also substantially down-regulated in the mutant striatum. In contrast, there was an increase in the number of striatal cholinergic interneurons. Down-regulated cortical GABA receptor and muscarinic receptor binding was also observed in addition to subtle morphological changes in the neuropeptide Y-expressing population of cortical neurons. The changes reflect the early cascade of events which follows the ablation of D1 dopamine receptor positive cells. Although extensive changes in a number of striatal and cortical neurons were demonstrated, only subtle transneuronal effects were seen in the nigrostriatal pathway. PMID- 10682710 TI - Late direct and transneuronal effects in mice with targeted expression of a toxin gene to D1 dopamine receptor neurons. AB - Detailed analysis of a novel transgenic model of basal ganglia disease has been undertaken. In this model the expression of an attenuated form of the diphtheria toxin gene was tightly controlled by D1 dopamine receptor regulatory domains. The behavioral and both direct toxin-mediated and transneuronal effects observed in pups in the first postnatal week have been described. Although younger pups are bradykinetic, older pups have a hyperkinetic syndrome with gait abnormality, postural instability and myoclonic jerks typical of human basal ganglia diseases such as Huntington's disease. As expected, striatal D1 dopamine receptor, dynorphin and substance P transcripts were not detected by in situ hybridization but there was a 27% increase in striatal D2 dopamine receptor messenger RNA and a 65% increase in enkephalin messenger RNA expression. Receptor autoradiographic studies confirmed the lack of D1-class binding in the mutant striatum and in contrast to young pups, a substantial increase in striatal D2-class binding. Autoradiographic quantitation also showed a 30% increase in striatal dopamine transporter binding. In addition to the changes described in the striatopallidal and nigrostriatal pathways, up-regulated dynorphin and substance P messenger RNA expression was also seen in the cortex. The capacity of the developing brain for neurochemical adaptation following injury is dramatic. The results show that primary loss of D1 dopamine receptor-positive striatonigral pathway neurons is sufficient to generate a hyperkinetic phenotype. PMID- 10682711 TI - Inhibitory control of the GABAergic transmission in the rat neostriatum by D2 dopamine receptors. AB - The aim of the study was to determine the role of dopamine on the GABAergic input to striatal projection neurons. Accordingly, the effect of the activation of dopamine D2-like receptors on GABA-mediated depolarizing postsynaptic potentials evoked in striatal slices by local stimulation was studied. Conventional intracellular recording techniques were used to record the synaptic responses. The experiments were done in the presence of 6-cyano-7-nitroquinoxaline-2,3-dione (20 microM) and (+)-2-amino-5-phosphonovaleric acid (40 microM) to block the participation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate/kainate and N-methyl-D-aspartate receptors in the synaptic response. The GABAergic nature of the response was assessed by its potentiation by pentobarbital (50 microM) and by its elimination by bicuculline or picrotoxin. At 100 nM, a concentration already maximal, dopamine inhibited by 55% the GABAergic synaptic response. The inhibitory effect was totally blocked by the selective antagonist of D2-like receptors, sulpiride (100 nM). The dopamine inhibition was observed only in one third of the studied neurons and was concentration dependent (IC50 = 14 nM). The inhibition was not associated with changes in the input resistance or any other membrane property. In addition, dopamine (50 nM) reduced the frequency but not the amplitude of spontaneous, bicuculline-sensitive depolarizing postsynaptic potentials. The D2-like receptor agonist quinpirole also dose-dependently (IC50 = 10 nM) inhibited the GABAergic synaptic response. As with dopamine, the inhibition did not change the membrane properties of the studied neurons. In addition, the quinpirole induced inhibition of the GABA response was accompanied by increased paired-pulse facilitation. The results indicate that D2-like receptors located on intrinsic GABAergic terminals in the rat striatum exert an inhibitory control of the GABAergic input to striatal projection neurons. The dopaminergic effect would be translated in facilitation of the firing of the neurons upon the arrival of the cortical input. PMID- 10682712 TI - Effect of acute and chronic administration of 1,2,3,4-tetrahydroisoquinoline on muscle tone, metabolism of dopamine in the striatum and tyrosine hydroxylase immunocytochemistry in the substantia nigra, in rats. AB - The effects of acute and chronic administration of 1,2,3,4 tetrahydroisoquinoline, an endogenous substance suspected of producing parkinsonism in humans, on the muscle tone and metabolism of dopamine in the striatum, and on the number of tyrosine hydroxylase-immunoreactive cells in the substantia nigra were investigated in rats. Muscle tone was examined using a combined mechanomyographic and electromyographic method which measured simultaneously the muscle resistance of the rat's hind foot to passive extension and flexion in the ankle joint and electromyographic activity of the antagonistic muscles of that joint: gastrocnemius and tibialis anterior. 1,2,3,4 Tetrahydroisoquinoline administered at doses of 50 and 100 mg/kg intraperitoneally for 19 days increased muscle resistance 1 h after the first injection (acute treatment), 1 h after the last injection (chronic treatment) and three days after compound withdrawal. Rigidity observed on the third day of 1,2,3,4-tetrahydroisoquinoline withdrawal was accompanied by an increased tonic (resting) electromyographic activity of the gastrocnemius and tibialis anterior muscles. At the same time, a significant reduction in the number of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra and a decrease in the dopamine level in the striatum were also found. A declining number of tyrosine hydroxylase-immunoreactive neurons in the whole substantia nigra showed a significant negative correlation with the enhanced muscle resistance, as well as with the tonic electromyographic activity recorded at rest, i.e. before the start of movements, from the gastrocnemius and tibialis anterior muscles. Our results suggest that 1,2,3,4-tetrahydroisoquinoline may be one of the endogenous substances involved in the progress of Parkinson's disease. PMID- 10682713 TI - Dopamine release and uptake are greater in female than male rat striatum as measured by fast cyclic voltammetry. AB - The present studies investigated sexual dimorphisms in dopamine release and uptake using fast-scan cyclic voltammetry in anesthetized rats and in brain slices. Electrical stimulation of the medial forebrain bundle of anesthetized rats at high frequency (60 Hz) elicited significantly more extracellular dopamine in the caudate nucleus of females than males. This sex difference was apparent over a range of current intensities applied to the stimulating electrode. Local electrical stimulation of brain slices in vitro verified in vivo results as more extracellular dopamine was elicited by single and 10 pulse stimulations in the caudate nucleus of females. Kinetic analysis of in vivo and in vitro dopamine overflow data indicated that dopamine release (the concentration of dopamine released per stimulus pulse) and the maximal velocity of dopamine uptake are greater in female rats, but the affinity of the transporter for dopamine was the same in males and females. None of these three parameters varied across the female estrous cycle. Linear regression analysis of dopamine release versus maximal uptake velocity data indicated a significant association of release and uptake sites in each sex and regression lines for males and females virtually overlapped. One explanation for these results is greater dopamine neuron terminal density in female caudate nucleus. These sexual dimorphisms in dopaminergic neurotransmission provide a novel, plausible mechanism to explain robust sex differences in behavioral responses of rats to psychostimulant drugs and may have implications for human neurological disorders and drug abuse. PMID- 10682714 TI - Lidocaine blockade of amygdala output in fear-conditioned rats reduces Fos expression in the ventrolateral periaqueductal gray. AB - We showed recently that conditioned fear to context induces Fos expression in the ventrolateral periaqueductal gray [Neuroscience (1997) 78, 165-177]. Neurons in this region are thought to play an important role in the expression of freezing during conditioned fear. To test the possibility that this activation comes directly from the amygdala, we looked at changes in Fos expression after a unilateral blockade of the ventral amygdalofugal pathway with lidocaine. The pathway contains fibres originating from the central nucleus of the amygdala that project directly and mainly ipsilaterally to the ventrolateral periaqueductal gray. Conditioned fear was evoked by re-exposing rats to the same box in which they had previously received electric footshocks. The test re-exposure was preceded by a unilateral microinjection of lidocaine (2%, 0.5-1 microl; n = 20) or saline (n = 14). Lidocaine was also tested in non-conditioned animals (n = 13). The results show that, when lidocaine was microinjected in the medial part of the central nucleus of the amygdala or along the ventral amygdalofugal pathway of conditioned rats, fear-induced Fos expression in the ventrolateral periaqueductal gray was reduced on the side ipsilateral to the injection (up to 37% reduction in comparison to the contralateral side). Ipsilateral reductions were also observed with saline, but they were weaker (maximum of 27% reduction). Fos expression remained low on both sides in the non-fear-conditioned animals injected with lidocaine. Finally, although freezing was only partly reduced in the conditioned animals unilaterally injected with lidocaine, it was significantly correlated to the ipsilateral reduction in Fos expression. This study provides direct evidence that the projection from the central nucleus of the amygdala to the ventrolateral periaqueductal gray is activated during fear and that it contributes to the Fos response of the ventrolateral periaqueductal gray. PMID- 10682715 TI - Correlations between serotonin level and single-cell firing in the rat's nucleus raphe magnus. AB - The relation between serotonin release and electrical activity was examined in the nucleus raphe magnus of rats anesthetized with pentobarbital. Serotonin levels were monitored through a carbon-fiber microelectrode by fast cyclic voltammetry (usually at 1 Hz). Single-cell firing was recorded through the same microelectrode, except during the voltammetry waveform and associated electrical artifact (totaling about 30 ms). Multi-barrel micropipettes incorporating the voltammetry electrode were used for iontophoresis of drugs. Cells were inhibited, excited or unaffected by noxious mechanical skin stimulation. These were respectively designated as off(M) cells, on(M) cells and neutral(M) cells, M denoting mechanical. During 3 min of pinching, serotonin slowly rose near seven of 14 on(M) cells and 26 of 46 off(M) cells; it fell near two off(M) cells; it was unchanged near all other cells, including six neutral(M) cells. On a finer spatiotemporal scale, near four of seven on(M) cells, 10 of 14 off(M) cells and 0 of four neutral(M) cells, average serotonin levels fell significantly within +/- 100 ms of spontaneous spikes. Lower serotonin may have caused the higher spike probability; the converse is theoretically unlikely, since delays between release and detection are estimated to exceed 100 ms. Increased serotonin and decreased firing were always seen following iontophoresis or intravenous injection (1 mg/kg) of the serotonin re-uptake inhibitor clomipramine (n = 7). Iontophoresis of +/- propranolol, whose serotonergic actions include antagonism and partial agonism at 5-HT1 receptors, also increased serotonin and decreased firing (n=4). Methiothepin (intravenous, 1 mg/kg), whose serotonergic actions include 5-HT1 and 5-HT2 antagonism, typically raised serotonin levels (four of five cells) and always blocked inhibition by clomipramine (n = 3). Iontophoresis of glutamate always lowered serotonin and increased firing (n = 4). Since serotonin levels and firing were usually inversely correlated, except near on(M) cells during pinch, we propose that serotonin is released from terminals of incoming nociceptive afferents. Prior neuroanatomical knowledge favors a midbrain origin for these afferents, while some of the drug findings suggest that their terminals possess inhibitory serotonergic autoreceptors, possibly of 5-HT1b subtype. The released serotonin could contribute to the inhibition of off(M) cells and excitation of on(M) cells by noxious stimulation, since inhibitory 5-HT1a receptors and excitatory 5-HT2 receptors, respectively, have previously been shown to dominate their serotonergic responses. PMID- 10682716 TI - Neurotrophin-induced rapid enhancement of membrane potential oscillations in mesencephalic trigeminal neurons. AB - We have proposed that neurotrophins, in addition to their trophic actions, act as neuromodulators in the adult central nervous system. As a first step to test this hypothesis, we examined in the adult rat slice preparation whether nerve growth factor and neurotrophin-3 are capable of altering the excitability of neurons of the mesencencephalic trigeminal nucleus. In contrast to vehicle pressure microapplication, which did not evoke changes in the electrophysiological properties of these neurons, neurotrophin application produced a significant increase in amplitude of the membrane potential oscillatory activity that is observed in these cells and a significant decrease in their threshold current. The latency of these effects ranged from 2 to 80 s and the duration ranged from 2 to 11 min. Neurotrophin-3 induced a decrease in input resistance and resting membrane potential in 58% of the cells; nerve growth factor induced a decrease in input resistance and resting membrane potential in 35% of the neurons. The spike configuration and action potential afterhyperpolarization potential remained unchanged following neurotrophin application. Tetrodotoxin blocked the membrane potential oscillatory activity of trigeminal mesencephalic neurons. Neurotrophin induced effects were not blocked by the tyrosine kinase inhibitor K-252a, whereas IgG-192, an antibody directed to the neurotrophin low-affinity receptor, enhanced excitability, as did neurotrophins. These results demonstrate that neurotrophins are capable of producing a rapid increase in the excitability of trigeminal mesencephalic neurons and suggest that their effects may be mediated by low affinity neurotrophin receptors. PMID- 10682717 TI - Repeated stimuli for axonal growth causes motoneuron death in adult rats: the effect of botulinum toxin followed by partial denervation. AB - Axons of motoneurons to tibialis anterior and extensor digitorum longus muscles of adult rats were induced to sprout by injecting botulinum toxin into them, by partial denervation or by a combination of the two procedures. Ten weeks later, the number of motoneurons innervating the control and operated tibialis anterior and extensor digitorum longus muscles was established by retrograde labelling with horseradish peroxidase. In the same preparations, the motoneurons were also stained with a Nissl stain (gallocyanin) to reveal motoneurons in the sciatic pool. Examination of the spinal cords from animals treated with botulinum toxin showed that the number of retrogradely labelled cells and those stained with gallocyanin in the ventral horn on the treated compared to the control side was unchanged. In rats that had their L4 spinal nerve sectioned on one side, the number of retrogradely labelled cells on the operated side was 48+/-3% (n = 5) of that present in the control unoperated ventral horn. Thus, just over half the innervation was removed by cutting the L4 spinal nerve. Counts made from gallocyanin-stained sections showed that 94+/-4% (n = 5) of motoneurons were present in the ventral horn on the operated side. Thus, section of the L4 spinal nerve did not lead to any death of motoneurons. In rats that had their muscles injected with botulinum toxin three weeks prior to partial denervation, the number of retrogradely labelled cells was reduced from 48+/-3% (n = 5) to 35+/-4% (n = 5). Moreover, only 67+/-5% (n = 5) of motoneurons stained with gallocyanin, suggesting that a proportion of motoneurons died after this combined procedure. This result was supported by experiments in which motor unit numbers in extensor digitorum longus muscles were determined by measurements of stepwise increments of force in response to stimulation of the motor nerve with increasing stimulus intensity. In partially denervated extensor digitorum longus muscles, 16.6+/-0.7 (n = 5) motor units could be identified, and in animals treated with botulinum toxin prior to partial denervation only 13.3+/-0.9 (n = 3) motor units were present. Taken together, these results show that treatment with botulinum toxin followed by partial denervation causes motoneuron death in adult rats. PMID- 10682718 TI - Identification of differentially expressed genes in dorsal root ganglia following partial sciatic nerve injury. AB - Partial sciatic nerve injury, a model of neuropathic pain, elicits a variety of neurochemical, electrophysiological and neuroanatomical changes in primary sensory neurons. We have used the technique of messenger RNA differential display to identify genes with altered expression in these neurons which may contribute to the development of aberrant sensation following such peripheral nerve damage. This approach identified 14 distinct complementary DNA clones, representing transcripts with increased ipsilateral expression in L4/5 dorsal root ganglia, two weeks after unilateral partial ligation of the rat sciatic nerve. Both Zucker diabetic fatty rats and their lean counterparts were used in this study but none of the transcripts identified showed an induction that was confined to one of the two groups. The majority of the clones did not show significant sequence similarity to previously reported genes and therefore may represent novel messenger RNA sequences or, alternatively, unknown regions of partially characterised messenger RNAs. Two of the clones represented transcripts for the known proteins muscle LIM protein and acidic epididymal glycoprotein, neither of which had previously been associated with expression in the nervous system. Reverse transcriptase-polymerase chain reaction analysis and in situ hybridization confirmed that the messenger RNA expression of both muscle LIM protein and acidic epididymal glycoprotein was induced in an ipsilateral-specific manner. Their localisations, examined with in situ hybridization in L5 dorsal root ganglia, were limited in each case to a sub-population of neuronal profiles. Those neuronal profiles that demonstrated muscle LIM protein hybridization were distributed across the profile size range, whereas the distribution of acidic epididymal glycoprotein-positive profiles appeared to be skewed towards smaller profiles. The induction of muscle LIM protein and acidic epididymal glycoprotein in dorsal root ganglia may play an important functional role in the adaptive response of primary sensory neurons following partial sciatic nerve injury. PMID- 10682719 TI - Calcium channels controlling acetylcholine release from preganglionic nerve terminals in rat autonomic ganglia. AB - Little is known about the nature of the calcium channels controlling neurotransmitter release from preganglionic parasympathetic nerve fibres. In the present study, the effects of selective calcium channel antagonists and amiloride were investigated on ganglionic neurotransmission. Conventional intracellular recording and focal extracellular recording techniques were used in rat submandibular and pelvic ganglia, respectively. Excitatory postsynaptic potentials and excitatory postsynaptic currents preceded by nerve terminal impulses were recorded as a measure of acetylcholine release from parasympathetic and sympathetic preganglionic fibres following nerve stimulation. The calcium channel antagonists omega-conotoxin GVIA (N type), nifedipine and nimodipine (L type), omega-conotoxin MVIIC and omega-agatoxin IVA (P/Q type), and Ni2+ (R type) had no functional inhibitory effects on synaptic transmission in both submandibular and pelvic ganglia. The potassium-sparing diuretic, amiloride, and its analogue, dimethyl amiloride, produced a reversible and concentration dependent inhibition of excitatory postsynaptic potential amplitude in the rat submandibular ganglion. The amplitude and frequency of spontaneous excitatory postsynaptic potentials and the sensitivity of the postsynaptic membrane to acetylcholine were unaffected by amiloride. In the rat pelvic ganglion, amiloride produced a concentration-dependent inhibition of excitatory postsynaptic currents without causing any detectable effects on the amplitude or configuration of the nerve terminal impulse. These results indicate that neurotransmitter release from preganglionic parasympathetic and sympathetic nerve terminals is resistant to inhibition by specific calcium channel antagonists of N-, L-, P/Q- and R-type calcium channels. Amiloride acts presynaptically to inhibit evoked transmitter release, but does not prevent action potential propagation in the nerve terminals, suggesting that amiloride may block the pharmacologically distinct calcium channel type(s) on rat preganglionic nerve terminals. PMID- 10682720 TI - Muscarinic receptor activation is a prerequisite for the endogenous release of nitric oxide modulating nicotinic transmission within the coeliac ganglion in the rabbit. AB - The aim of the present study was to investigate whether the activation of muscarinic receptors is a preliminary step to the endogenous release of nitric oxide modulating nicotinic transmission within the prevertebral ganglia. This work has been performed in vitro in isolated rabbit coeliac ganglion. The electrical activity of the ganglionic neurons was recorded using intracellular recording techniques. When a train of pulses of supramaximal intensity was applied to the splanchnic nerves, gradual depression of fast nicotinic transmission occurred: the pulses do not systematically elicit action potentials, but very often elicit excitatory postsynaptic potentials only. The use of pharmacological agents that interfere with the nitric oxide pathway such as L arginine (precursor of nitric oxide) or 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (nitric oxide scavenger) demonstrated that nitric oxide modulates this depression phenomenon by facilitating or inhibiting the nicotinic transmission of the ganglionic neurons. A nitric oxide donor (diethylamine/nitric oxide complex) induced an inhibition of the nicotinic synaptic transmission. In the presence of the muscarinic receptors antagonist atropine, L-arginine and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1 oxyl-3-oxide failed to modify the nicotinic transmission of the ganglionic neurons but diethylamine/nitric oxide complex was still able to inhibit it. These results demonstrate that in the coeliac ganglion, the activation of muscarinic cholinergic receptors is a prerequisite for the activation of neuronal nitric oxide synthase in preganglionic fibres. The nitric oxide released then exerts a facilitation or an inhibition of the nicotinic transmission of the ganglionic neurons. Atropine triggered a facilitation of the nicotinic transmission when superfused alone and an inhibition when superfused in the presence of 2-(4 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide. These results confirm that muscarinic receptors activate the nitric oxide pathway modulating the nicotinic transmission of the prevertebral neurons. Our results also demonstrate that when the nitric oxide pathway is blocked, activation of muscarinic receptors leads to facilitation of the nicotinic transmission. Our study brings new insights concerning the modulation by nitric oxide and by muscarinic receptors of the synaptic transmission within the prevertebral ganglia. PMID- 10682721 TI - Localization of metabotropic glutamate receptor type 2 in the human brain. AB - Metabotropic glutamate receptors are a heterogeneous family of G-protein-coupled receptors that are linked to multiple second messenger systems to regulate neuronal excitability and synaptic transmission. To elucidate the physiological role of these receptors in human central nervous system function and dysfunction at the receptor protein level requires the use of selective antibodies to determine the phenotype of cells expressing particular receptor subtypes. To this end the present study has examined the regional and cellular localization of the metabotropic glutamate type 2 receptor protein in selected human brain regions. After epitope prediction, antibodies have been generated against a short synthetic peptide corresponding to amino acid residues located in the putative intracellular carboxy-terminus and subsequently applied to an immunohistochemical investigation. Antibodies specifically detected the type 2 receptor in transfected mammalian cells and also recognized a major band of 98,000 mol. wt in western blots of human brain tissue membranes. At the light microscope level immunohistochemical studies have demonstrated that type 2-like immunoreactivity was widely distributed in the human brain, being characterized by the presence of a strong immunoreaction in multiple cortical regions, and in structures comprising the basal ganglia, to include the caudate nucleus, putamen, globus pallidus, substantia nigra and subthalamic nucleus. In the hippocampal formation, immunoreactivity was predominant in selective cell layers of both the dentate gyrus and cornu ammonis, the subicular complex and entorhinal cortex. In the thalamus, multiple subnuclei showed reaction product. In the cerebellar cortex, immunoreactivity was expressed in a number of cell layers and cell types. Furthermore, using double immunofluorescence we confirmed that the type 2 receptor is a product of normal resting astrocytes in the cerebral cortex in particular. This antibody provides a new immunological tool with the potential to evaluate the distribution of human metabotropic glutamate receptor 2 protein in other brain regions and in human central nervous system diseases. PMID- 10682722 TI - Involvement of caspase-1 proteases in hypoxic brain injury. effects of their inhibitors in developing neurons. AB - To further explore the contribution of caspase-1/interleukin-1beta-convening enzyme in the consequences of hypoxia in developing brain neurons, its temporal expression profile was analysed by immunohistochemistry and western blotting in cultured neurons from the embryonic rat forebrain subjected to a hypoxic stress (95% N2/5% CO2 for 6 h), and proteolytic activity of caspase-1 was monitored as a function of time by measuring the degradation of a selective colorimetric substrate (N-acetyl-Tyr-Val-Ala-Asp-p-nitroanilide). In addition, the influence of pre- and posthypoxic treatments by caspase-1 inhibitors (N-acetyl-Tyr-Val-Ala Asp-aldehyde and N-acetyl-Tyr-Val-Ala-Asp-chloromethylketone) was tested on cell outcome. Hypoxia led to delayed apoptotic neuronal death, with an elevation of the expression of both pro-caspase-1 and caspase-1 active cleavage product (ICE p20) for up to 96 h after cell reoxygenation. As reflected by cleavage of the specific substrate, caspase-1 activity progressively increased between 24 h and 96 h posthypoxia, and was blocked by inhibitors in a dose-dependent fashion. The inhibitory compounds, including when given 24 h after hypoxia, prevented neuronal death, reduced apoptosis hallmarks and also increased the number of mitotic neurons, suggesting they might promote neurogenesis. Similar observations were made when neurons were exposed to a sublethal hypoxia (i.e. 3 h). These data emphasize the participation of caspase-1 in neuronal injury consecutive to oxygen deprivation, and provide new insight into the possible cellular mechanisms by which caspase inhibitors may protect developing brain neurons. PMID- 10682723 TI - Sites of action of thyrotropin-releasing hormone on central nervous system neurons revealed by expression of the immediate-early gene c-fos in the rat. AB - Centrally administered thyrotropin-releasing hormone produces a number of physiological and behavioral changes, e.g., a general antidepressant effect, increasing body temperature, and elevated blood pressure. However, the specific brain sites of action responsible for the centrally activating property of thyrotropin-releasing hormone have not been precisely determined. Using chloral hydrate-anesthetized adult Sprague-Dawley rats, we compared the distribution of Fos-like immunoreactivity after intracerebroventricular administration of thyrotropin-releasing hormone with the results after intracerebroventricular injection of vehicle alone. Some rats were paralysed and artificially ventilated to avoid possible Fos expression secondarily induced by autonomic (e.g., respiratory) disturbances. In thyrotropin-releasing hormone administered rats, selective Fos-like immunoreactivity was observed in V/VI layers of the pre- and infralimbic areas of the medial prefrontal cortex, the ventral midline thalamus, and the nucleus of the solitary tract as well as in the adjacent reticular formation. Fos-like immunoreactivity was significantly reduced in most areas of the cerebral cortex (II/III layers), the shell of the nucleus accumbens, the medial amygdaloid nucleus, parts of the hypothalamus, and the periaqueductal gray. These data suggest that various behavioral and autonomic responses induced by centrally administered thyrotropin-releasing hormone might be produced through the complex neural circuitry comprising the above structures, which are presumed to be implicated in limbic and/or autonomic functions. PMID- 10682724 TI - Repair of articular cartilage defects one year after treatment with recombinant human bone morphogenetic protein-2 (rhBMP-2). AB - BACKGROUND: Damaged articular cartilage has a limited ability to repair. Operative removal of damaged cartilage and penetration into the subchondral bone to allow population of the defect with progenitor cells can result in filling of the defect with repair tissue. However, this repair tissue often degenerates over time because of its inability to withstand the mechanical forces to which it is subjected. We previously reported that recombinant human bone morphogenetic protein-2 (rhBMP-2) improves the repair of full-thickness defects of cartilage as long as six months postoperatively. We have now extended that study to examine the quality of the repair tissue at one year. METHODS: Full-thickness defects of cartilage were created in the trochlear groove of twenty-five adult New Zealand White rabbits. Eight defects were left empty, eight were filled with a collagen sponge, and nine were filled with a collagen sponge impregnated with five micrograms of rhBMP-2. The animals were killed at fifty-two weeks postoperatively, and the gross appearance of the healed defect was assessed. The repair tissue was examined histologically and was evaluated, according to a grading scale, by four individuals who were blinded with respect to the treatment. The tissue sections were immunostained with antibodies against type-I collagen, type-II collagen, aggrecan, and link protein. The residence time of the rhBMP-2 in the cartilage defect was evaluated in vivo with use of scintigraphic imaging of radiolabeled protein. RESULTS: One year after a single implantation of a collagen sponge containing five micrograms of rhBMP-2, the defects had a significantly better histological appearance than the untreated defects (those left empty or filled with a collagen sponge). The histological features that showed improvement were integration at the margin, cellular morphology, architecture within the defect, and reformation of the tidemark. The total scores were also better for the defects treated with rhBMP-2 than for the untreated defects, but in no instance was the repair tissue identical to normal articular cartilage. The thickness of the cartilage in the defects treated with rhBMP-2 was 70 percent that of the normal cartilage, an observation that was identical to that at twenty-four weeks postoperatively. Immunostaining demonstrated significantly less type-I collagen in the defects treated with rhBMP-2 than in the untreated defects. Immunostaining for other matrix components showed no difference among the treatment groups. The mean residence time of rhBMP-2 in the cartilage defects was eight days with an elimination half-life of 5.6 days. Detectable amounts of rhBMP-2 were present as long as fourteen days after implantation. CONCLUSIONS: The problems associated with operative repair of cartilage include the formation of fibrocartilage rather than normal articular cartilage and the degeneration of that repair tissue over time. Our results demonstrate that the addition of rhBMP-2 to the operative site after creation of a full-thickness defect results in an improvement in the histological appearance and composition of the extracellular matrix at one year postoperatively. If these experimental results translate directly to the clinical situation, it is possible that the addition of rhBMP-2 to existing operative treatments for the repair of cartilage may improve the repair process and may help to maintain the integrity of the repair tissue. PMID- 10682725 TI - Ciprofloxacin inhibition of experimental fracture healing. AB - BACKGROUND: Fluoroquinolones, such as ciprofloxacin, have an adverse effect on growing cartilage and endochondral ossification in children. This study was carried out to determine whether ciprofloxacin also has an adverse effect on the healing of experimental fractures. METHODS: Sixty male 300-gram Wistar rats were divided equally into three groups, which received ciprofloxacin, cefazolin, or no treatment for three weeks, beginning seven days after production of a closed, nondisplaced, bilateral femoral fracture. The serum concentrations of the ciprofloxacin and the cefazolin were 2.4 and 146 micrograms per milliliter, respectively. Radiographic, histological, and biomechanical studies were used to evaluate fracture-healing. RESULTS: Radiographs revealed significantly more advanced healing of the control fractures compared with the fractures in the ciprofloxacin-treated group (average stage, 2.1 compared with 1.5, p = 0.01). The cefazolin-treated group was not different from the controls with respect to radiographic healing (average stage, 1.8 compared with 2.1, p = 0.18). Torsional strength-testing of fracture callus exposed to ciprofloxacin revealed a 16 percent decrease in strength compared with the controls (284 compared with 338 newton-millimeters, p = 0.04) and a 49 percent decrease in stiffness (twenty compared with thirty-nine newton-millimeters per degree, p = 0.001). The biomechanical strength in the cefazolin-treated group was not different from that of the controls. Fracture calluses in the animals treated with ciprofloxacin showed abnormalities in cartilage morphology and endochondral bone formation and a significant decrease in the number of chondrocytes compared with the controls (0.77 x 10(4) compared with 1.3 x 10(4) cells per square millimeter, p = 0.004). CONCLUSIONS: These data suggest that experimental fractures exposed to therapeutic concentrations of ciprofloxacin in serum demonstrate diminished healing during the early stages of fracture repair. The administration of ciprofloxacin during early fracture repair may compromise the clinical course of fracture-healing. PMID- 10682726 TI - Joint angular velocity in spastic gait and the influence of muscle-tendon lengthening. AB - BACKGROUND: Joint angular velocity (the rate of flexion and extension of a joint) is related to the dynamics of muscle activation and force generation during walking. Therefore, the goal of this research was to examine the joint angular velocity in normal and spastic gait and changes resulting from muscle-tendon lengthening (recession and tenotomy) in patients who have spastic cerebral palsy. METHODS: The gait patterns of forty patients who had been diagnosed with spastic cerebral palsy (mean age, 8.3 years; range, 3.7 to 14.8 years) and of seventy three age-matched, normally developing subjects were evaluated with three dimensional motion analysis and electromyography. The patients who had cerebral palsy were evaluated before muscle-tendon lengthening and nine months after treatment. RESULTS: The gait patterns of the patients who had cerebral palsy were characterized by increased flexion of the knee in the stance phase, premature plantar flexion of the ankle, and reduced joint angular velocities compared with the patterns of the normally developing subjects. Even though muscle-tendon lengthening altered sagittal joint angles in gait, the joint angular velocities were generally unchanged at the hip and knee. Only the ankle demonstrated modified angular velocities, including reduced dorsiflexion velocity at foot strike and improved dorsiflexion velocity through mid-stance, after treatment. Electromyographic changes included reduced amplitude of the gastrocnemius-soleus during the loading phase and decreased knee coactivity (the ratio of quadriceps and hamstring activation) at toe-off. Principal component analyses showed that, compared with joint-angle data, joint angular velocity was better able to discriminate between the gait patterns of the normal and cerebral palsy groups. CONCLUSIONS: This study showed that muscle-tendon lengthening corrects biomechanical alignment as reflected by changes in sagittal joint angles. However, joint angular velocity and electromyographic data suggest that the underlying neural input remains largely unchanged at the hip and knee. Conversely, electromyographic changes and changes in velocity in the ankle indicate that the activation pattern of the gastrocnemius-soleus complex in response to stretch was altered by recession of the complex. PMID- 10682727 TI - Electromyographic and gait analysis of forty-three patients after rotationplasty. AB - BACKGROUND: Rotationplasty is considered to be a treatment option for patients who have had a primary malignant bone tumor of the distal part of the femur or the proximal part of the tibia. The present study was performed to evaluate the muscle activity, the kinetics (range of motion of the hip and knee joints), and the kinematics (joint moments) after rotationplasty and to determine whether there was an association between these parameters and the functional outcome. METHODS: Forty-three patients who had been managed with rotationplasty for the treatment of a femoral or tibial bone tumor were evaluated clinically and functionally. The mean age (and standard deviation) at the time of follow-up was 24.4 +/- 10.7 years (range, eight to sixty-eight years), the mean age at the time of the procedure was 17.8 +/- 10.2 years (range, seven to sixty-three years), and the mean duration of follow-up was 6.7 +/- 4.9 years (range, 0.7 to eighteen years). Instrumented gait and electromyographic analyses were performed. The qualitative data were compared with the functional outcome, which was determined with the functional evaluation score of the Musculoskeletal Tumor Society. RESULTS: Gait analysis revealed a fairly normal walking pattern with a slight limp and a lateral lean of the trunk over the ipsilateral limb that led to a reduced joint moment in the hip (moment on involved side, 68 percent [compared with a control group]; moment on uninvolved side, 81 percent). The ranges of motion of the hips (uninvolved side, 42.0 +/- 8.2 degrees; involved side, 42.4 +/ 8.0 degrees) and the knees (uninvolved side, 59.7 +/- 5.0 degrees; involved side [former ankle joint], 58.1 +/- 11.6 degrees) were symmetrical even though the knee-motion pattern of the involved limb indicated a slightly reduced extensor mechanism in 51 percent (twenty-two) and a markedly reduced extensor mechanism in 35 percent (fifteen) of the forty-three patients. Electromyography revealed function of the muscles of the involved limb, with comparable amplitudes in the involved and uninvolved limbs. The leg muscles of the involved limb were active in the stance phase (the soleus and the lateral and medial heads of the gastrocnemius) and the swing phase (the peroneus longus and the tibialis anterior) according to their function in relation to the new knee joint. The patients had a good functional result, with a mean score of 23.9 +/- 2.7 of 30 points. With the numbers available for study, we could not show the duration of follow-up to be related to the overall outcome, but the age at the time of the operation was related to the total functional score as well as to gait and walking ability (p < 0.05). CONCLUSIONS: The results of the electromyographic and gait analyses demonstrated good functional restoration of gait following rotationplasty. PMID- 10682728 TI - Acetabular revision with use of a bilobed component inserted without cement in patients who have acetabular bone-stock deficiency. AB - BACKGROUND: Massive deficiency of acetabular bone stock is a challenging problem in the increasing number of patients who need a revision of a failed hip arthroplasty. The bilobed cup has been presented as one alternative reconstruction technique for hips with extensive acetabular bone loss. The purpose of this study was to assess the results with use of a bilobed acetabular component inserted without cement for revision reconstruction in hips with acetabular bone deficiency in order to clarify the indications for its use and to identify the factors that influence the clinical and radiographic outcome. METHODS: Forty-one hips in thirty-eight patients had an acetabular revision with a bilobed acetabular component inserted without cement between December 1991 and December 1995. These hips were a subset of the 414 hips treated with an acetabular revision during the same period of time. One patient was lost to follow-up, and one died during the study period. Two patients who could not return for radiographic evaluation completed questionnaires. The remaining thirty four patients (thirty-seven hips) were evaluated radiographically and clinically and were followed for an average of forty-one months (range, twenty-four to sixty six months). RESULTS: Radiographic analysis demonstrated an improvement in the average vertical displacement of the hip center. At the time of the latest follow up examination, 76 percent (twenty-eight) of the thirty-seven cups were stable, 8 percent (three) were probably unstable with a change in the screw position but no definite migration of the cup, and 16 percent (six) were unstable. Eight of the nine loose or probably loose components were in patients who had more than two centimeters of superior migration of the component and disruption of Kohler's line on preoperative radiographs. Additionally, implants were more likely to become unstable (demonstrating more than 4 degrees of change in the abduction angle or more than four millimeters of radiographic migration) when the inferior aspect of the component did not extend to or distal to the interteardrop line, which indicated that the component was undersized. CONCLUSIONS: On the basis of our early rate of probable or definite loosening of 24 percent (nine of thirty seven cups) and the technical difficulties involved, we do not recommend the routine use of this component. We believe that this device is indicated when a patient has an oblong-shaped acetabular defect and the surgeon wants to correct an elevated hip center. However, the medial wall of the acetabulum (Kohler's line) should be intact if the failed component has migrated more than two centimeters. An alternative reconstruction technique, such as use of a structural allograft with or without an acetabular cage, is also an option in this situation. PMID- 10682729 TI - Anticoagulant treatment of thromboembolism with intravenous heparin therapy in the early postoperative period following total joint arthroplasty. AB - BACKGROUND: Treatment of thromboembolism with intravenous heparin therapy in the early postoperative period after total joint arthroplasty has been associated with a high rate of complications. The purpose of the present study was to compare the rate of bleeding complications in a group of patients who required intravenous heparin therapy for the treatment of thromboembolism after total hip or knee arthroplasty with the rate in a control group of patients who received only prophylactic anticoagulation. METHODS: The postoperative courses of forty four consecutive patients who were managed with intravenous administration of heparin and oral administration of warfarin for the treatment of a thromboembolic event following unilateral total hip or knee arthroplasty were compared with those of a control group of 376 consecutive patients who had these same procedures but did not have a thromboembolic complication. The patients in the control group were managed with prophylactic anticoagulation with use of enoxaparin. Sixty-eight percent (thirty) of the forty-four patients in the heparin group received the initial dose of heparin on or before the fourth postoperative day, and 82 percent (thirty-six) received an initial bolus of 5000 units of heparin at the initiation of therapy. RESULTS: The rate of bleeding complications was 9 percent (four of forty-four) in the heparin group, compared with 6 percent (twenty-three of 376) in the control group (p = 0.44). The mean transfusion requirement in the heparin group (1.8 units of packed red blood cells) was significantly greater than that in the control group (0.8 unit) (p < 0.0001). Three of the four patients who had a bleeding complication while receiving heparin and warfarin had coagulation parameters that were substantially higher than recommended levels. The mean duration of hospitalization in the heparin group (fifteen days) was significantly longer than that in the control group (seven days) (p < 0.0001). CONCLUSIONS: The results of the present study suggest that the use of intravenous heparin therapy for the treatment of thromboembolism in the early postoperative period after total joint arthroplasty is associated with a rate of bleeding complications that is similar to that associated with the use of prophylactic anticoagulation with use of enoxaparin alone. One should expect an increased transfusion requirement and a longer duration of hospitalization for patients who require intravenous heparin therapy for the treatment of a thromboembolic event. PMID- 10682730 TI - Bilateral stress fractures of the anterior part of the tibial cortex. A case report. PMID- 10682731 TI - A transverse acetabular nonunion treated with computer-assisted percutaneous internal fixation. A case report. PMID- 10682732 TI - Displaced intra-articular fractures of the calcaneus. PMID- 10682733 TI - Recent advances in venous thromboembolic prophylaxis during and after total hip replacement. PMID- 10682734 TI - Design issues in clinical studies of the in vivo volumetric wear rate of polyethylene bearing components. PMID- 10682735 TI - Current concepts review. Internet resources for orthopaedic surgeons. PMID- 10682736 TI - Survival analysis of hips treated with core decompression or vascularized fibular grafting because of avascular necrosis. PMID- 10682737 TI - Survival analysis of hips treated with core decompression or vascularized fibular grafting because of avascular necrosis. PMID- 10682738 TI - Survival analysis of hips treated with core decompression or vascularized fibular grafting because of avascular necrosis. PMID- 10682739 TI - Acetabular involvement in osteonecrosis of the femoral head. PMID- 10682740 TI - Clinical and pathological findings of a newly recognized disease of elephants caused by endotheliotropic herpesviruses. AB - The unique clinical and pathological findings in nine Asian (Elephas maximus) and two African (Loxodonta africana) elephants from North American Zoos with a highly fatal disease caused by novel endotheliotropic herpesviruses are described. Identification of the viruses by molecular techniques and some epidemiological aspects of the disease were previously reported. Consensus primer polymerase chain reaction (PCR) combined with sequencing yielded molecular evidence that confirmed the presence of two novel but related herpesviruses associated with the disease, one in Asian elephants and the second in African elephants. Disease onset was acute, with lethargy, edema of the head and thoracic limbs, oral ulceration and cyanosis of the tongue followed by death of most animals in 1 to 7 days. Pertinent laboratory findings in two of three clinically evaluated animals included lymphocytopenia and thrombocytopenia. Two affected young Asian elephants recovered after a 3 to 4 wk course of therapy with the anti-herpesvirus drug famciclovir. Necropsy findings in the fatal cases included pericardial effusion and extensive petechial hemorrhages in the heart and throughout the peritoneal cavity, hepatomegaly, cyanosis of the tongue, intestinal hemorrhage, and ulceration. Histologically, there were extensive microhemorrhages and edema throughout the myocardium and mild, subacute myocarditis. Similar hemorrhagic lesions with inflammation were evident in the tongue, liver, and large intestine. Lesions in these target organs were accompanied by amphophilic to basophilic intranuclear viral inclusion bodies in capillary endothelial cells. Transmission electron microscopy of the endothelial inclusion bodies revealed 80 to 92 nm diameter viral capsids consistent with herpesvirus morphology. The short course of the herpesvirus infections, with sudden deaths in all but the two surviving elephants, was ascribed to acute cardiac failure attributed to herpesvirus induced capillary injury with extensive myocardial hemorrhage and edema. PMID- 10682742 TI - Seroepidemiology of chlamydial infections of wild ruminants in Spain. AB - Chlamydial infections were determined serologically among wild ruminants in the Nature Park of the Sierras de Cazorla, Segura y Las Villas (CNP; Spain). Sampling was done during the period from 1990-95. There were 1,244 blood samples collected, consisting of 490 from fallow deer (Dama dama), 343 from mouflon (Ovis mussimon), 283 from red deer (Cervus elaphus) and 128 from Spanish ibex (Capra pyrenaica). Specific complement-fixing antibodies of Chlamydia spp. were detected by means of microtechnique, using lipopolysaccharide antigen. The relationship of biological (species, sex, age), temporal (year) and territorial (central and peripheral areas) factors to seropositive prevalence was examined, and preliminary data were collected on whether or not sheep and goat herds grazing in the peripheral areas of the park also were infected with Chlamydia spp. Chlamydiosis was common in the four species of wild ruminants in the CNP in all the years studied. The prevalence of Chlamydia sp. in mouflon (37%) was significantly greater than in fallow deer (30%), and both had a significantly higher prevalence rate than Spanish ibex and red deer (both 24%). The four species of wild ruminants were similar in that they act as reservoirs of Chlamydia spp., although their receptivity may be different, and the infection can certainly be maintained among these animals by intra-group transmission. The differences in prevalences and geometric mean titers (GMT), both between the sexes (male versus female) and between different ages (adult versus juvenile), were insignificant in all four species. For all species of wild ruminants both prevalence rates and GMTs were greater in populations occupying the peripheral areas of the park than in those inhabiting the central area. Herds of sheep and goats had a high prevalence of chlamydiosis. Intertransmission of Chlamydia sp. between wild and domestic ruminants occurred through grazing on the same pastures. The highest mean prevalence (44%) of patent infections (CFT titers of > or =1:80) was detected in red deer, although this frequency was not significantly different from those observed in mouflon (39%), Spanish ibex (38%), and fallow deer (37%). The proportion of patent infection was higher in females than in males, and none of the juveniles (<2-yr-old) showed patent infections. The prevalence of predicted patent chlamydial infections was always higher in the peripheral areas of the park, although only among mouflon and fallow deer were the differences statistically significant. PMID- 10682741 TI - Role of peridomestic birds in the transmission of St. Louis encephalitis virus in southern California. AB - In response to the 1984 St. Louis encephalitis (SLE) epidemic in the Los Angeles Basin of southern California (USA), an investigative program was initiated to evaluate the interactive components of the SLE virus transmission cycle. From 1987 through 1996 (10 yr), 52,589 birds were bled and their sera tested for SLE and western equine encephalomyelitis (WEE) virus antibodies by the hemagglutination inhibition (HAI) test. Eighty-three percent of the birds tested were house finches (Carpodacus mexicanus) (48.7%) and house sparrows (Passer domesticus) (34.6%); 1.1% of these birds were positive for SLE antibodies. Prevalence of WEE antibodies was negligible. The analysis of 5,481 sera from rock doves (Columbia livia) yielded 3.6% SLE positives and 0.4% WEE positives. Collection sites were maintained as study sites when identified as positive bird, mosquito, and SLE virus activity localities; others were abandoned. Serial serum samples from 7,749 banded house sparrows and 9,428 banded house finches from these selected sites demonstrated year-round SLE virus transmission. One location exhibited significant numbers of house finches undergoing annual SLE seroconversion and a number of seroconversion-reversion-reconversion sequences suggesting either viral reinfection from mosquitoes or recrudescence by latent virus. A proportion of both bird species also lived for longer than 1 yr, thus, increasing the possibility of virus carry-over from autumn to spring. Assessment of concurrently collected mosquitoes indicated no correlative association between mosquito populations and SLE seroconversion and reconversion. European house sparrows introduced in the 1800's may have provided a supplemental link to the existing SLE virus enzootic cycle involving endemic house finches. Meteorological factors are reviewed as possible important correlates of SLE epidemics. The house finch/house sparrow serosurveillance system is also evaluated for use as an "Early Warning" indicator of SLE virus activity. PMID- 10682743 TI - Characterization of Lyme disease spirochetes isolated from ticks and vertebrates in North Carolina. AB - Borrelia burgdorferi isolates obtained from numerous locations and from different hosts in North Carolina, were compared to previously characterized strains of the Lyme disease spirochete and other Borrelia spp. The spirochete isolates were confirmed to be B. burgdorferi sensu stricto based on immunofluorescence (IFA) using a monoclonal antibody to outer surface protein A (Osp A [H5332]) and polymerase chain reaction (PCR) using a species-specific nested primer for a conserved region of the gene that encodes for flagellin. In addition, the isolates tested positive in Western blots with species-specific monoclonal antibodies for outer surface protein A and OspB (84c), and the genus-specific, monoclonal antibody to flagellin (H9724). Infectivity studies with several of these isolates were conducted using Mus musculus and Oryzomys palustris and the isolates exhibited markedly different levels of infectivity. This study demonstrates that B. burgdorferi sensu stricto is present and naturally transmitted on the Outer Banks and in the Coastal Plain and Piedmont regions of North Carolina. PMID- 10682744 TI - Mange epizootic in white-nosed coatis in western Mexico. AB - From November of 1994 to June of 1996 an epizootic of mange, probably caused by the mite Notoedres cati, occurred in white-nosed coatis (Nasua narica) in the tropical dry forests of the Chamela-Cuixmala Biosphere Reserve in western Mexico. A monitoring scheme to determine the extent and severity of the epizootic within coatis was implemented. Trapping periods and transects were conducted for 2 yr. To control the spread of the disease, all captured infected coatis were either euthanized or treated with acaricides such as Butox and Ivomec-F, depending on the severity of their infection. Four other species of wild mammals and feral cats had skin conditions resembling mange. A more severe problem with the disease was predicted and later confirmed in the less isolated areas of the reserve, with a higher density of coatis. Our results indicate that epizootics may be more prone to occur in areas with greater fragmentation and less isolation from anthropogenic influence. Interestingly, although there was an apparently severe impact of the mange epizootic in the coati population, the long-term impact of the disease is unknown but appears to be negligible. So in order to understand the role of diseases in wildlife populations, long-term experimental studies are required. PMID- 10682745 TI - Trichomoniasis in a Bonelli's eagle population in Spain. AB - During 1980-97, trichomoniasis was detected in nestlings of Bonelli's eagle Hieraaetus fasciatus in Catalonia (Spain). In 1993 Trichomonas gallinae was isolated in 36% of nestlings (n = 39) and affected 41% of broods (n = 22). Overall, trichomoniasis was one of the most important single nestling mortality factor, accounting for 22% of total chick mortality, and causing the death of 2% of chicks. Trichomoniasis deaths took place during the second half of the nestling period. The median age at death was 45.5 days. Although the presence of the parasite was not related to the composition of the diet or parental age, pairs that developed the disease ate more pigeons and included more often non adult birds. At present trichomoniasis apparently has little demographic impact on the Bonelli's eagle population in Catalonia, but the eventual spread of this disease in chicks and its unknown effects on adults might be of concern. PMID- 10682746 TI - Detection, identification, and correction of a bias in an epidemiological study. AB - The relative lack of epidemiological studies of natural populations is partly due to the difficulty of obtaining samples that are both large enough and representative of the population. Here, we present the result of an epidemiological study (December 1992-August 1995) of feline immunodeficiency virus (FIV) in a free-roaming population of domestic cats (Felis catus), with a special emphasis on sample bias. Over five trapping periods, the prevalence of FIV in sampled cats steadily declined. Across these samples we consistently achieved a very large sampling fraction (approximately 60% of the population), the sex ratio, age and weight distributions remained stable with time in the samples, and the sex ratio was similar in the samples and the population. These indices would normally indicate that our samples were representative, suggesting the decline in FIV prevalence to be real. However, a concomitant ecological study of the whole population revealed an important bias in the samples, with an initial high probability of capturing a few individuals, which appeared significantly more likely to be FIV-infected, and then a lower probability of recapturing them. Since our protocol resulted in a non-random sampling, subsequent trappings were designed to avoid this bias, by also capturing individuals who had previously learned to escape capture. This modified capture regime revealed that FIV prevalence was in fact constant in the population. This study shows how samples of large size, which are stable and appear representative of the population, can still be biased. These results may have major implications for other studies based on trapping. PMID- 10682747 TI - Effects of delivery method on serological responses of bighorn sheep to a multivalent Pasteurella haemolytica supernatant vaccine. AB - The safety and efficacy of a remotely delivered multivalent Pasteurella haemolytica supernatant vaccine (serotypes A2 and T10) were examined in captive Rocky, Mountain bighorn sheep (Ovis canadensis canadensis). Twenty bighorn sheep were grouped according to baseline leukotoxin neutralizing antibody titers (< or =2 or >2 log2(-1)) and vaccination history (previously vaccinated or unvaccinated). Within these groups, animals were randomly assigned to one of two delivery treatments: hand injection (control) or biobullet implantation. All bighorns received a single dose from the same lot of vaccine (n = 10/treatment); four additional animals were injected intramuscularly with 0.9% saline as unvaccinated sentinels. Mild, transient lameness one day after hand injection or biobullet implantation was the only adverse effect. Serum neutralizing antibody titers to P. haemolytica leukotoxin differed between delivery treatments (P = 0.009) and among baseline titer/vaccination history groups (P = 0.013). Neutralizing titers were higher among hand-injected bighorns. Although neutralizing titers were lower among implanted bighorns than hand-injected controls at 1 wk (P = 0.002) and 2 wk (P = 0.021) after vaccination, seroconversion rates in response to implantation (6/10) and hand injection (9/10) did not differ (P = 0.303). Agglutinating antibody titers to T10 were high and did not vary over time or between delivery treatments. Agglutinating antibody titers to A2 in the hand-injected controls were not different (P > or = 0.07) than those in bighorns vaccinated with biobullet implantation. These data demonstrate that although hand injection elicits higher absolute titers, biobullet implantation may also stimulate effective antibody responses to P. haemolytica supernatant vaccine. Further evaluation of biobullet vaccination against pneumonic pasteurellosis in free-ranging populations of wild bighorn sheep is warranted. PMID- 10682748 TI - A modified bait for oral delivery of biological agents to raccoons and feral swine. AB - A field study was conducted on Ossabaw Island (Georgia, USA) in March 1994 to evaluate four different types of bait for delivering orally effective biological agents to raccoons (Procyon lotor) and feral swine (Sus scrofa). A deep-fried corndog batter bait, which was previously shown to be ingested by both captive and free-ranging raccoons, and a polymer fishmeal bait which had been shown effective for both raccoons and feral swine were compared with a grain-based dog food meal polymer bait topically coated with corn oil and cornmeal or with fish oil and fishmeal. Tracking stations were used to determine the number of each bait type visited and removed by animals visiting stations. We found no significant differences in the numbers of different baits removed by either species. These data support the results of earlier studies which also indicated that an inexpensive grain-based matrix bait surface-coated with attractive flavors can be used to deliver oral biologics to problem species. PMID- 10682749 TI - Effect of jaw shape in kill-traps on time to loss of palpebral reflexes in brushtail possums. AB - The effect of three configurations of the jaw of kill-traps on time to loss of palpebral reflex of brushtail possums (Trichosurus vulpecula) was assessed. Traps were Standard, with an offset rotating jaw closing past a pear-shaped constriction, Offset, with a rotating jaw closing past a straight edge, and Opposing, with a rotating jaw closing directly onto a static bar. Possums captured in the Standard and Offset traps had significantly lower times to loss of palpebral reflexes (42 and 50 sec respectively) than those captured in Opposing traps (122 sec). Both the Standard and Offset traps achieved total occlusion of both carotid arteries more frequently than the Opposing trap. Thus the killing effectiveness of kill traps designed for capturing possums can be improved by offsetting the jaws without the need to increase the power of the trap. Therefore, for some target species, such modifications might satisfy the demands of animal welfare proponents for traps that kill rapidly, without compromising trapper safety. PMID- 10682750 TI - Scurvy in capybaras bred in captivity in Argentine. AB - In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity. PMID- 10682752 TI - Influence of diet on the hematology and serum biochemistry of zinc-intoxicated mallards. AB - Changes in hematological and serum biochemistry parameters in female zinc (Zn) dosed farm-raised mallards (Anas platyrhynchos) fed four different diets were examined. Sixty ducks received an average dose of 0.97 g of Zn in the form of eight, 3.30-mm diameter shot pellets containing 98% Zn and 2% tin, and another 60 ducks were sham-dosed as controls. Fifteen ducks from each of the two dosing groups were assigned to one of four dietary treatments: corn only, corn with soil, commercial duck ration only, or commercial duck ration with soil. Shot pellet dissolution rates ranged from 7 mg/Zn/day to 27 mg/Zn/day. Regardless of diet, the Zn dose resulted in mortality; incoordination; paralysis and anorexia; decreased body, liver, pancreas, gonad, and gizzard weight; increased kidney weight; and macroscopic lesions. Zn-dosed ducks had a lower mean erythrocyte packed cell volume (PCV), higher mean reticulocyte count, and a greater number of individuals with immature and/or abnormal erythrocytes, than did control mallards. Mean total leucocyte counts were higher in Zn-dosed ducks than in controls. Zn-dosed ducks that had soil available had higher leucocyte counts than those without soil. Zn-dosed ducks were characterized by a marked heterophilia and relative lymphopenia. In Zn-dosed ducks, the mean lymphocyte count was highest in those provided a commercial duck ration, and lowest in those fed corn. In control ducks, the mean lymphocyte count was highest in ducks fed corn, and lowest in those provided soil along with a commercial duck ration. Zn-dosed mallards had higher serum aspartate aminotransferase and amylase levels, and lower alkaline phosphatase activities than control ducks. Serum phosphorus and uric acid concentrations were higher, and calcium, glucose, and total protein levels lower, in Zn-dosed ducks than in control ducks. Diet did affect serum calcium, phosphorus, total protein, and uric acid concentrations. Differences in erythrocyte and leucocyte parameters, serum enzyme activities, and metabolite concentrations were associated with dose and diet effects. Diets high in protein and other organic matter and calcium and phosphorus did not prevent or substantially alleviate Zn toxicosis in farm-raised mallard ducks. PMID- 10682751 TI - Biochemical responses to fibropapilloma and captivity in the green turtle. AB - Blood biochemical parameters were compared for green turtles (Chelonia mydas) with and without green turtle fibropapillomatosis (GTFP) from both captive and wild populations in Hawaii (USA) and from a captive population from California (USA), during the period between 1994 and 1996. Statistical analysis did not detect an influence of disease in any of the blood parameters for free-ranging turtles; however, captive turtles in Hawaii with GTFP had significantly higher levels of alkaline phosphatase and significantly lower levels of lactate compared to non-tumored captive turtles. Multivariate analysis found that biochemical profiles could be used to accurately predict if turtles were healthy or afflicted with GTFP. Discriminant function analysis correctly classified turtles as being with or without GTFP in 89% of cases, suggesting that diseased animals had a distinct signature of plasma biochemistries. Measurements of blood parameters identified numerous differences between captive and wild green turtles in Hawaii. Levels of corticosterone, lactate, triglyceride, glucose, and calcium were significantly higher in wild green turtles as compared to captive turtles, while uric acid levels were significantly lower in wild turtles as compared to captive turtles. Additionally, turtles from Sea World of California (San Diego, California, USA), which had been in captivity the longest, had higher levels of alanine aminotransferase and triglycerides as compared to nearly all other groups. Differences in diet, sampling methods, environmental conditions, and turtle size, help to interpret these results. PMID- 10682753 TI - Immobilization of California sea lions using medetomidine plus ketamine with and without isoflurane and reversal with atipamezole. AB - The use of medetomidine and ketamine, alone and in combination with isoflurane, with atipamezole reversal was evaluated for immobilizing 51 California sea lions (Zalophus californianus) for a variety of medical procedures at a rehabilitation center in northern California (USA) between May 1997 and August 1998. Animals were given 140 microg/kg medetomidine with 2.5 mg/kg ketamine intramuscularly. Mean (+/-SD) time to maximal effect was 8+/-5 min. At the end of the procedure, animals were given 200 microg/kg atipamezole intramuscularly. Immobilization and recovery times were, respectively, 25+/-12 and 9+/-7 min for 35 animals maintained with medetomidine and ketamine alone and 58+/-30 and 9+/-9 min for 16 animals intubated and maintained with isoflurane. No mortalities occurred as a result of the immobilizations. Disadvantages of the medetomidine and ketamine combination included a moderate variation in time to maximal effect and plane of sedation, a large injection volume and high cost. However, this combination offers safe and reversible immobilization that can be easily administered by the intramuscular route and that produces a plane of anesthesia that is sufficient to carry out most routine diagnostic procedures. PMID- 10682755 TI - Pathology of experimental toxoplasmosis in eastern barred bandicoots in Tasmania. AB - Wild-caught eastern barred bandicoots (Perameles gunnii) initially seronegative to Toxoplasma gondii, were inoculated orally with approximately 100 T. gondii oocysts. The bandicoots were maintained in indoor pens under laboratory conditions and observed daily. Serial blood samples were tested for agglutinating antibodies to T. gondii. Inoculated bandicoots died 15 and 17 days post infection. A rise in Direct Agglutination Test (DAT) titres was detected at the time of death (1:256, 1:64 respectively). Clinical observations, serological changes, gross findings at necropsy, and histopathological changes were consistent with acute toxoplasmosis. The findings indicate that eastern barred bandicoots are likely to die from primary T. gondii infection, often even before detectable antibodies are produced, reinforcing the significance of toxoplasmosis as a potential contributor to the reduction in numbers of wild populations of eastern barred bandicoots. PMID- 10682754 TI - Immobilization of free-ranging nine-banded and great long-nosed armadillos with three anesthetic combinations. AB - Nine-banded (n = 47) and great (n = 31) long-nosed armadillos (Dasypus novemcinctus and Dasypus kappleri) were immobilized for clinical examination and collection of biological samples as part of a wildlife rescue during the filling of a hydroelectric dam (Petit Saut, French Guiana) from May 1994 to April 1995. Three intramuscular (i.m.) anesthetic combinations were evaluated: (1) tiletamine/zolazepam (T/Z) at a dose of 8.5 mg/kg in 12 nine-banded long-nosed armadillos (NBA) and 10 great long-nosed armadillos (GLA), (2) ketamine at 40 mg/kg combined with xylazine at 1.0 mg/kg (K/X) in 18 NBA and nine GLA, and (3) ketamine at 7.5 mg/kg combined with medetomidine at 75 microng/kg (K/M) in 17 NBA and 12 GLA, antagonized by 375 microg/kg atipamezole. Induction was smooth, ranged from mean +/- SD = 2.8+/-0.6 to 4.3+/-1.8 min, and did not differ significantly between protocols, species, or sex. In NBA, immobilization time ranged from 43.8+/-27.8 to 66.5+/-40.0 min and did not differ between protocols or sex. Muscle relaxation was judged to be better with K/X and K/M versus T/Z. In GLA, the response to the anesthetic protocols was more variable and immobilization time ranged from 30.4+/-6.2 to 98.4+/-33.7 min. The main difference was observed in GLA females receiving the T/Z combination, in which immobilization time was significantly longer versus males, but also versus GLA K/M group, and versus NBA T/Z group. Effects on body temperature, heart rate and respiratory rate were limited. Thirty six to 50% of the individuals showed hypoxemia (SpO2 < 85%) throughout anesthesia and values <80% also were recorded but the hypoxemia was not associated with clinical signs. With T/Z and K/X, recovery was irregular and prolonged up to 2 to 3 hr in some individuals. In K/M groups, first standing was observed 1.0 to 16.4 min after i.m. atipamezole injection without adverse effects. Finally, the three anesthetic combinations used in this study were effective and safe agents for 30 to 40 min immobilizations including minor surgery procedures. The ability to antagonize the medetomidine-induced sedation with atipamezole significantly reduces the recovery time, making the K/M combination preferable, especially in field conditions. PMID- 10682756 TI - Earthworms as paratenic hosts of toxoplasmosis in eastern barred bandicoots in Tasmania. AB - An experimental feeding study was designed to assess the role of earthworms in the transmission of Toxoplasma gondii infection to eastern barred bandicoots (Perameles gunnii). Six animals with no agglutinating antibodies to T. gondii were fed artificially cultured earthworms that had been maintained in autoclaved nutrient-enriched soil. Two animals were given earthworms that had been maintained in soil contaminated with T. gondii oocysts (P89/VEG strain); two animals were fed on earthworms, which initially had been exposed to soil containing T. gondii oocysts then transferred through three changes of sterile soil; two control bandicoots were fed earthworms maintained in sterile soil. Both bandicoots fed earthworms maintained in T. gondii contaminated soil died 11 and 14 days after feeding. The necropsy findings were consistent with acute toxoplasmosis. Bandicoots fed earthworms exposed to oocysts but then transferred through changes of sterilized soil remained healthy as did control animals. All surviving animals remained seronegative over the 6 wk observation period after feeding. These findings confirm that earthworms, a major component of the natural diet of P. gunnii, can transmit T. gondii infection. It appears that oocysts present in the alimentary tracts of the worms, rather than infective stages of T. gondii in worm somatic tissues, are responsible for these infections. PMID- 10682757 TI - Naturally occurring and experimentally transmitted Hepatozoon americanum in coyotes from Oklahoma. AB - Twenty free-ranging coyotes (Canis latrans) in Oklahoma (USA) were examined for the presence of naturally occurring infections with Hepatozoon americanum and to determine if bone lesions attributable to H. americanum were present. Although eight of the 20 free-ranging coyotes were found to be naturally infected with H. americanum, no bone lesions were detected. In addition, two coyote pups were exposed to H. americanum oocysts collected from experimentally infected ticks and the course of the resulting infection was followed. Both experimentally infected coyotes developed hepatozoonosis detectable by specific muscle lesions beginning 4 wk after exposure. Bone lesions were detected grossly and histologically at necropsy. Histologic evidence of periosteal bone proliferation ranged from segmental areas of plump hypercellularity and thickening of the periosteum, with minor degrees of osteogenesis, to extensive proliferation of woven bone and periosteal hypercellularity and thickening. Nymphal Amblyomma maculatum that fed on one of the experimentally infected coyote pups became infected and mature H. americanum oocysts were recovered when the ticks molted to adults. These results demonstrate that coyotes in some parts of Oklahoma are naturally infected with H. americanum, that experimentally infected coyotes can develop clinical disease, including characteristic bone lesions, and that A. maculatum nymphs can acquire infections by feeding on them. PMID- 10682758 TI - Louse flies on birds of Baja California. AB - Louse flies were collected from 401 birds of 32 species captured in autumn of 1996 in Baja California Sur (Mexico). Only one louse fly species (Microlynchia pusilla) was found. It occurred in four of the 164 common ground doves (Columbina passerina) collected. This is a new a host species for this louse fly. PMID- 10682759 TI - Prevalence of Bartonella henselae antibody in Florida panthers. AB - Serum samples from 28 free-ranging Florida panthers (Puma concolor coryi) and seven mountain lions from Texas (P. concolor stanleyana) living in south Florida (USA) between 1997 to 1998 were tested for antibodies to Bartonella henselae. Twenty percent (7/35) of the samples were reactive to B. henselae antisera with a subspecies prevalence of 18% (5/ 28) for Florida panthers and 28% (2/7) for cougars from Texas (USA). There was not a significant sex related difference in infection rates among the Florida panthers. Antibody prevalence was higher in panthers <2-yr of age (40%) compared to panthers >2-yr (13%). Compared to studies of antibody prevalence in mountain lions (P. concolor) from California (USA), overall seroprevalence was lower as was prevalence in panthers >2-yr-old. However, the seroprevalence in animals <2-yr from southern Florida was similar to prevalences reported in mountain lions or domestic felids in California. PMID- 10682760 TI - Antibodies to selected disease agents in translocated wild turkeys in California. AB - Wild turkeys (Meleagris gallopavo) trapped within California (n = 715) or imported into California from other states (n = 381) from 1986 to 1996 were tested for exposure to certain disease agents. Prevalence of antibody to Mycoplasma gallisepticum, Mycoplasma meleagridis, Salmonella pullorum, Salmonella typhimurium, Newcastle disease virus, and avian influenza virus was low (0-4%) for wild turkeys trapped within California. With the exception of antibody prevalence to M. meleagridis of 33%, the same was true for wild turkeys imported into California from other states. Antibody prevalence to Mycoplasma synoviae was 8-10% for both groups. PMID- 10682761 TI - Typing of pestiviruses from eland in Zimbabwe. AB - Pestiviruses were isolated from three eland (Taurotragus oryx) in Zimbabwe. The viruses were characterised by typing with monoclonal antibodies and by partial genetic sequencing. All were similar to bovine viral diarrhea viruses commonly isolated from cattle. This suggests that bovine viral diarrhea virus can spread from cattle to eland. PMID- 10682762 TI - Rabies in an American bison from North Dakota. AB - In North Dakota (USA) during April 1998, a ranched female bison (Bison bison) was found dead. At gross necropsy, there was profound hair loss and consolidated lung lobes. Intracytoplasmic neuronal inclusions suggestive of Negri bodies were observed in the brain stem and hippocampus, and a diagnosis of rabies was confirmed by the fluorescent antibody test. Antigenic typing demonstrated the occurrence of a rabies virus variant associated with skunks from the upper midwestern USA. This case of a rabid bison was one of only four such instances recorded from the USA over the past 40 yr, and is the first case report of rabies in a bison that reports clinical, pathologic, and antigenic findings. Although rabies in bison is rare, veterinarians and wildlife managers that work closely with such non-traditional species are reminded of the dangers that zoonoses such as rabies present. PMID- 10682763 TI - Effect of climate and type of storage container on aflatoxin production in corn and its associated risks to wildlife species. AB - The effects of grain storage containers on aflatoxin production, and the relationship between the level of aflatoxin and the number and weight of fluorescing kernels were determined in corn (Zea maize) stored in controlled climate regimes. Two hundred and forty 100-g samples were held up to 3 mos using four types of storage containers placed in four climates. Storage containers included corn placed in metal cans, paper bags, plastic bags, and paper bags placed in plastic bags. Climates were constant during the duration of the project and included a combination of temperatures and humidities. Temperatures were 29 32 C and 14-18 C; relative humidities were 85-88% and 35-40%. In addition, corn was exposed to environmental conditions conductive for aflatoxin production and 100 g samples were randomly collected, examined under ultraviolet light for fluorescence, and then quantified for aflatoxin levels. Corn samples tested negative for aflatoxin at the beginning of the project. Main (i.e., container, climate, and month) and interactive effects were not observed. Mean levels of aflatoxin ranged from 0 to 151 microg/kg. Aflatoxin was produced regardless of type of storage container, time of storage, and climatic conditions; however, only 8% of the samples produced aflatoxin levels that exceeded 50 microg/kg. Fluorescing corn ranged from 0 to 19 kernels per sample, while aflatoxin levels ranged from 0 to 1,375 microg/kg for the same samples. No relationships were found between the number and weight of fluorescing kernels of corn and aflatoxin levels. The black light test yielded a false negative rate of 23% when in fact the aflatoxin concentrations exceeded 50 microg/kg. Therefore, quantifying fluorescing grain under UV light should not be considered a feasible alternative for aflatoxin testing of grain intended for wildlife. PMID- 10682764 TI - Lead poisoning in a northern bobwhite in Georgia. AB - A northern bobwhite (Colinus virginianus) was observed with partial paralysis on 3 March 1997 and found dead on 8 March 1997 on Di-Lane Plantation Wildlife Management Area (Burke County, Georgia, USA). The juvenile male was necropsied by the Southeastern Cooperative Wildlife Disease Study (Athens, Georgia) and diagnosed with lead toxicosis. The bobwhite had liver tissue lead levels of 399 parts per million wet weight and two worn 1-mm diameter lead shot pellets were found in the gizzard. PMID- 10682765 TI - Desert bighorn sheep mortality due to presumptive type C botulism in California. AB - During a routine telemetry flight of the Mojave Desert (California, USA) in August 1995, mortality signals were detected from two of 12 radio-collared female desert bighorn sheep (Ovis canadensis) in the vicinity of Old Dad Peak in San Bernardino County (California). A series of field investigations determined that at least 45 bighorn sheep had died near two artificial water catchments (guzzlers), including 13 bighorn sheep which had presumably drowned in a guzzler tank. Samples from water contaminated by decomposing bighorn sheep carcasses and hemolyzed blood from a fresh bighorn sheep carcass were tested for the presence of pesticides, heavy metals, strychnine, blue-green algae, Clostridium botulinum toxin, ethylene glycol, nitrates, nitrites, sodium, and salts. Mouse bioassay and enzyme-linked immunosorbent assay detected type C botulinum toxin in the hemolyzed blood and in fly larvae and pupae. This, coupled with negative results from other analyses, led us to conclude that type C botulinum poisoning was most likely responsible for the mortality of bighorn sheep outside the guzzler tank. PMID- 10682766 TI - Letter to the editor concerning Arnold-Chiari malformation in a captive African lion cub. PMID- 10682767 TI - Double pylorus sign. PMID- 10682768 TI - Minimally invasive treatment of malignant hepatic tumors: at the threshold of a major breakthrough. AB - Six existing minimally invasive techniques for the treatment of primary and secondary malignant hepatic tumors--radio-frequency ablation, microwave ablation, laser ablation, cryoablation, ethanol ablation, and chemoembolization--are reviewed and debated by noted authorities from six institutions from around the world. All of the authors currently believe that surgery remains the treatment of choice for patients with resectable hepatic tumors. However, the clinical results of each of the minimally invasive techniques presented have exceeded those obtained with conventional chemotherapy or radiation therapy. Thus, for nonsurgical patients, these techniques are becoming standard independent or adjuvant therapies. In addition, with continued improvement in technology and increasing clinical experience, one or more of these minimally invasive techniques may soon challenge surgical resection as the treatment of choice for patients with limited hepatic tumor. PMID- 10682769 TI - CT and MR imaging findings of bowel ischemia from various primary causes. AB - Ischemic bowel disease represents a broad spectrum of diseases with various clinical and radiologic manifestations, which range from localized transient ischemia to catastrophic necrosis of the gastrointestinal tract. The primary causes of insufficient blood flow to the intestine are diverse and include thromboembolism, nonocclusive causes, bowel obstruction, neoplasms, vasculitis, abdominal inflammatory conditions, trauma, chemotherapy, radiation, and corrosive injury. Computed tomography (CT) or magnetic resonance (MR) imaging can demonstrate the ischemic bowel segment and may be helpful in determining the primary cause. The CT and MR imaging findings include bowel wall thickening with or without the target sign, intramural pneumatosis, mesenteric or portal venous gas, and mesenteric arterial or venous thromboembolism. Other CT findings include engorgement of mesenteric veins and mesenteric edema, lack of bowel wall enhancement, increased enhancement of the thickened bowel wall, bowel obstruction, and infarction of other abdominal organs. However, regardless of the primary cause, the imaging findings of bowel ischemia are similar. Furthermore, the bowel changes simulate inflammatory or neoplastic conditions. Understanding the pathogenesis of various conditions leading to mesenteric ischemia helps the radiologist recognize ischemic bowel disease and avoid delayed diagnosis, unnecessary surgery, or less than optimal management. PMID- 10682770 TI - Solitary pulmonary nodules: Part I. Morphologic evaluation for differentiation of benign and malignant lesions. AB - The solitary pulmonary nodule is a common radiologic abnormality that is often detected incidentally. Although most solitary pulmonary nodules have benign causes, many represent stage I lung cancers and must be distinguished from benign nodules in an expeditious and cost-effective manner. Evaluation of specific morphologic features of a solitary pulmonary nodule with conventional imaging techniques can help differentiate benign from malignant nodules and obviate further costly assessment. Small size and smooth, well-defined margins are suggestive of but not diagnostic for benignity. Lobulated contour as well as an irregular or spiculated margin with distortion of adjacent vessels are typically associated with malignancy. There is considerable overlap in the internal characteristics (eg, attenuation, cavitation, wall thickness) of benign and malignant nodules. The presence of intranodular fat is a reliable indicator of a hamartoma. The presence and pattern of calcification can also help differentiate benign from malignant nodules. Computed tomography (CT) (particularly thin section CT) is 10-20 times more sensitive than standard radiography and allows objective, quantitative assessment of calcification. Initial evaluation often results in nonspecific findings, in which case nodules are classified as indeterminate and require further evaluation to exclude malignancy. Growth rate assessment, Bayesian analysis, contrast material-enhanced CT, positron emission tomography, and transthoracic needle aspiration biopsy can be useful in this regard. PMID- 10682771 TI - Solitary pulmonary nodules: Part II. Evaluation of the indeterminate nodule. AB - Various strategies may be used to evaluate indeterminate solitary pulmonary nodules. Growth rate assessment is an important and cost-effective step in the evaluation of these nodules. Clinical features (eg, patient age, history of prior malignancy, presenting symptoms, smoking history) can be useful in suggesting the diagnosis and aiding in management planning. Bayesian analysis allows more precise determination of the probability of malignancy (pCa). Decision analysis models suggest that the most cost-effective management strategy depends on the pCa for a given nodule. At contrast material-enhanced computed tomography, nodular enhancement of less than 15 HU is strongly predictive of a benign lesion, whereas enhancement of more than 20 HU typically indicates malignancy. At 2 [fluorine-18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography, lesions with low FDG uptake are typically benign, whereas those with increased FDG uptake are typically malignant. Results of transthoracic needle aspiration biopsy influence management in approximately 50% of cases and, in indeterminate lesions with a pCa between 0.05 and 0.6, is the best initial diagnostic procedure. It is optimally used in peripheral nodules and has been reported to establish a benign diagnosis in up to 91% of cases. Although there is no one correct management approach, the ability to distinguish benign from malignant solitary pulmonary lesions has improved with the use of these strategies. PMID- 10682772 TI - Unusual radiologic findings in the thorax after radiation therapy. AB - Radiation therapy is used to treat many intrathoracic and chest wall malignancies. A variety of changes may occur after radiation therapy to the thorax. Radiation therapy produces dramatic effects in the lung. Pulmonary necrosis is an uncommon, severe, late complication of adjuvant postoperative radiation therapy. Bronchiolitis obliterans with organizing pneumonia is a distinct clinicopathologic entity characterized by patchy, migratory, peripheral air-space infiltrates. Radiation therapy can also cause spontaneous pneumothorax, mesothelioma, and lung cancer. In the mediastinum, radiation therapy may cause thymic cysts, calcified lymph nodes, and esophageal injuries. Cardiovascular complications of radiation therapy are often delayed and insidious. Premature coronary artery stenosis occurs after radiation therapy to the mediastinum. Radiation therapy may also give rise to calcifications of the ascending aorta, pericardial disease, valvular injuries, and conduction abnormalities. Women who undergo thoracic irradiation before the age of 30 years have a high risk of developing a second breast cancer. Radiation-induced sarcomas are an infrequent but well-recognized complication of radiation therapy. Other chest wall injuries due to radiation therapy are osteochondroma and rib or clavicle fractures. Knowledge of the imaging features of injuries caused by radiation therapy can prevent misinterpretation as recurrent tumor and may facilitate further treatment. PMID- 10682773 TI - Back to basics: keeping patients and families in the loop. PMID- 10682774 TI - Radiation-induced lung disease and the impact of radiation methods on imaging features. AB - Although radiologic findings in radiation-induced lung disease are well described in the literature, the influence exerted on these findings by different radiation methods is not well understood. Radiation treatment of non-small cell lung cancer varies depending on the location and extent of disease. Irradiation with oblique beam angles results in unusual distribution of radiation-induced lung disease. Small cell lung cancer is treated with irradiation concurrent with or following chemotherapy, and portal arrangements are controversial. In breast cancer, use of tangential beam portals may induce radiation pneumonitis or fibrosis at the peripheral lung anterolaterally. Use of supraclavicular portals may produce lesions in the lung apex that appear similar to pulmonary tuberculosis. In esophageal cancer, radiation portals with a 5-6-cm margin above and below the tumor are generally recommended, and computed tomography (CT) frequently demonstrates radiation-related lung damage adjacent to the mediastinum. In mediastinal tumors, the mantle field includes all the major lymph node regions above the diaphragm. Radiation pneumonitis varies from minimal to extremely marked change in the paramediastinal areas and in both apices. CT is more sensitive to radiation-induced lung disease than chest radiography and demonstrates related changes earlier. Furthermore, it more clearly depicts the precise distribution and pattern of disease. Familiarity with the imaging findings in radiation-induced lung disease produced by different radiation methods will help radiologists interpret abnormalities seen at chest radiography and CT in affected patients. PMID- 10682775 TI - Review of criteria appropriate for a very low probability of pulmonary embolism on ventilation-perfusion lung scans: a position paper. AB - The "low-probability" interpretation of ventilation-perfusion lung scans has been characterized as misleading or even dangerous because of the high prevalence of pulmonary embolism associated with such an interpretation. Since the completion of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) study, analyses of the PIOPED database have allowed identification of several abnormalities seen on ventilation-perfusion scans that have a positive predictive value (PPV) for pulmonary embolism of less than 10%. These include nonsegmental perfusion abnormalities (PPV = 8%), perfusion defects smaller than the corresponding areas of increased opacity at chest radiography (PPV = 8%), matched ventilation-perfusion abnormalities in two or three zones of a single lung (PPV = 3%), one to three small segmental perfusion defects (PPV = 1%), triple matched defects in the upper or middle lung zone (PPV = 4%), and the stripe sign (PPV = 7%). Use of these abnormalities as interpretative criteria constitutes "very low probability" interpretation and will reduce the number of low-probability interpretations of ventilation-perfusion lung scans, which may be considered nondiagnostic because of the unacceptably high rate of false-negative results. This will enhance the utility of the ventilation-perfusion lung scan for screening patients with suspected pulmonary embolism. PMID- 10682776 TI - MR angiography of the thoracic aorta with an electrocardiographically triggered breath-hold contrast-enhanced sequence. AB - An electrocardiographically (ECG) triggered breath-hold contrast material enhanced magnetic resonance (MR) angiography sequence has been developed for imaging the thoracic aorta. A three-dimensional (3D) gradient-echo sequence is used with a contrast material bolus. Forty-nine patients with various aortic abnormalities and five healthy volunteers underwent imaging with the sequence. All studies were performed in a single breath hold. ECG-triggered breath-hold contrast-enhanced MR angiography was tolerated in 48 of the 49 patients. The images demonstrated no respiratory motion artifacts and diminished pulsation artifacts. The cardiac chambers, aortic root, ascending and descending aorta, aortic arch, proximal arch vessels, and proximal coronary arteries were clearly demonstrated and not obscured by ghost artifacts. The 3D data set allowed excellent multiplanar reformation, permitting orthogonal or oblique views of the vascular anatomy. A variety of congenital and acquired abnormalities were clearly identified. When this sequence is used, it is important to evaluate both the maximum-intensity projection and source images. Delayed imaging should be performed to detect late filling. In conjunction with cine MR and T1-weighted spin-echo imaging, ECG-triggered breath-hold contrast-enhanced MR angiography should be considered the technique of choice for imaging the thoracic aorta. PMID- 10682777 TI - Optimized diagnostic angiography in high-risk patients with severe peripheral vascular disease. AB - Conventional arteriography remains the usual method for preoperative assessment of severe peripheral vascular disease (PVD). Unfortunately, many peripheral arteriograms are still performed with a suboptimal technique, which can cause significant diagnostic errors in patients with severe PVD. A suboptimal technique may be due to poor collimation (causing incorrect exposure and incorrect gray scale), excessive patient-film distance (magnification unsharpness), inadequate volume or density of contrast material, poor contrast resolution (screen-film arteriography), nonselective injection, patient movement, and pressure from restraints or incorrect patient position (failure to profile lesions, pseudo occlusion from external pressure or plantar flexion). The technique of selective digital subtraction arteriography (DSA) allows one to avoid these errors. The superior contrast resolution of DSA allows use of lower concentrations of contrast material. Selective injection into the external iliac artery allows proper positioning and improves image quality. Demonstration of distal vessels is best achieved by using biplane arteriography. For patients with severe resting ischemia, especially those with diabetes, high-quality selective DSA is essential to ensure that all distal vessels suitable for distal bypass grafting are identified. When properly performed, selective DSA remains the investigation of choice for reliably demonstrating arterial anatomy in high-risk patients with severe PVD. PMID- 10682778 TI - Dynamic subtraction contrast-enhanced MR angiography: technique, clinical applications, and pitfalls. AB - Rapid advances in techniques of contrast material-enhanced magnetic resonance (MR) angiography have enabled evaluation of the entire aorta and the main arteries. Dynamic subtraction MR angiography consists of first-pass imaging of long segments of arteries by using a three-dimensional fast field echo sequence with multiple rapid bolus injections of a small dose of gadopentetate dimeglumine. Subtraction enables clear demonstration of the enhanced vascular lumen by eliminating background signal. Improved temporal resolution and repeated sequences after gadopentetate dimeglumine administration allow demonstration of arteries and veins separately. Double subtraction postprocessing can be used to eliminate arterial enhancement in demonstration of the portal and systemic veins. Additional postprocessing can be used to demonstrate arteries in a single image in patients with aortic dissection or a prolonged circulation time. To optimize the examination, the pulse sequence, injection dose, injection rate, timing of the start of data acquisition, imaging time, breath holding, section thickness, and coil selection should be considered. This technique is flexible enough to be applied in a variety of clinical settings, including atherosclerotic occlusive disease, aneurysm of aortoiliac arteries, bypass graft, Takayasu arteritis, aortic dissection, antiphospholipid antibody syndrome, renal artery disease, pelvic vascular disease, and the portomesenteric venous system. PMID- 10682779 TI - Pediatric voiding cystourethrography: a pictorial guide. AB - Voiding cystourethrography is commonly performed in children with prenatally diagnosed hydronephrosis, urinary tract infections, and voiding abnormalities. Voiding cystourethrography can be performed with many variations designed to optimize visualization of disease and minimize radiation exposure. The procedure should include assessment of the spine and pelvis; masses or opaque calculi; bladder capacity, contour, and emptying capability; presence and grade of reflux; and urethral appearance. Radiologists differ as to whether the patient should void prior to catheterization. Anteroposterior imaging of the bladder is performed during early filling; little or no imaging is necessary during intermediate filling. When bladder filling is complete, steep oblique images that are centered on the ureterovesical junction should be obtained. If reflux is observed, the ipsilateral renal fossa may be imaged prior to voiding. With a smaller than expected voiding volume, bladder refilling is recommended. Voiding around the catheter is also strongly recommended. In girls, one anteroposterior image of the urethra is usually sufficient; in boys, the entire urethra must be imaged. Steep oblique imaging is optimal. At the conclusion of voiding, each renal fossa should be imaged to detect reflux missed at fluoroscopy as well as other anomalies. Familiarity with these abnormalities and use of proper techniques will allow detection of most common pathologic conditions with very low radiation exposure. PMID- 10682780 TI - US approach to jaundice in infants and children. AB - High-resolution real-time ultrasonography (US) serves as an important tool for differentiation of obstructive and nonobstructive causes of jaundice in infants and children, independent of liver function. Unconjugated hyperbilirubinemia occurs in approximately 60% of normal term infants and in 80% of preterm infants. Persistence of neonatal jaundice beyond 2 weeks of age demands US evaluation to differentiate between the three most common causes: hepatitis, biliary atresia, and choledochal cyst. In all three conditions, the hepatic echotexture is diffusely coarse and hyperechoic, but this appearance may be seen in a variety of hepatic inflammatory, obstructive, and metabolic processes. Thus, hepatic scintigraphy and at times percutaneous liver biopsy are necessary to narrow the differential diagnosis and to identify patients who require more invasive techniques (eg, intraoperative cholangiography). US is useful for demonstrating inspissated bile and biliary duct stones. In infants, stones are usually secondary to obstructive congenital anomalies of the biliary tract, total parenteral nutrition, furosemide treatment, phototherapy, dehydration, infection, hemolytic anemia, and short-gut syndrome, whereas in older children, stones are usually associated with sickle cell disease, bowel resection, hemolytic anemia, and choledochal cyst. Jaundice in infants and children may also be due to cirrhosis, benign strictures, and neoplastic processes. PMID- 10682781 TI - Renal lymphoma: CT patterns with emphasis on helical CT. AB - Renal lymphoma is most often seen in conjunction with multisystemic, disseminated lymphoma or as tumor recurrence. Renal lymphoma may also be seen in immunocompromised patients or, rarely, as primary disease. Computed tomography (CT) is the most sensitive, efficient, and comprehensive examination for evaluation of the kidneys in patients with suspected renal lymphoma. Helical CT in particular improves detection and characterization of lymphomatous renal involvement by optimizing contrast dynamics and data acquisition and is the current modality of choice for accurate staging of lymphoma. Typical CT patterns in renal lymphoma include single and multiple masses, invasion from contiguous retroperitoneal disease, perirenal disease, and diffuse renal infiltration. Atypical CT patterns may also be encountered and provide a diagnostic challenge. These include spontaneous hemorrhage, necrosis, heterogeneous attenuation, cystic transformation, and calcification. Solid renal masses including renal cell carcinoma and metastases are the most commonly encountered entities that mimic renal lymphoma at CT and require biopsy for definitive diagnosis. CT (particularly helical CT) is useful in the evaluation of patients with suspected renal lymphoma, and familiarity with the spectrum of findings in renal lymphoma is important for accurate diagnosis. PMID- 10682782 TI - From the archives of the AFIP. Infiltrative renal lesions: radiologic-pathologic correlation. Armed Forces Institute of Pathology. AB - Most renal masses exhibit an expansile growth pattern characterized by radial tumor enlargement that displaces normal renal parenchyma and forms spherical, often exophytic, lesions. These expansile masses have pushing margins that impress adjacent normal renal parenchyma but do not infiltrate it; this behavior results in a well-defined, encapsulated appearance at both radiologic and gross pathologic examination. In contrast, certain disease processes involve the kidney in an infiltrative fashion by using the normal renal architecture as scaffolding for interstitial growth. These infiltrative renal lesions lack a sharp border of demarcation with the normal parenchyma and therefore demonstrate ill-defined zones of transition between the lesion and normal parenchyma. Although infiltrative lesions frequently enlarge the kidney, its reniform shape is usually maintained. Cross-sectional imaging can often help distinguish infiltrative from expansile growth patterns through analysis of the parenchymal interface between the process and the kidney, the effect of the lesion on the collecting system and renal sinus, and the overall renal morphology. A wide variety of neoplastic and inflammatory conditions characteristically involve the kidney by cellular infiltration. Although considerable overlap of the imaging features exists among the various infiltrative processes, the correct diagnosis may be suspected when the clinical data and associated radiologic findings are considered together. PMID- 10682783 TI - Image interpretation session: 1999. Intraductal mucin-producing tumor of the pancreas. PMID- 10682784 TI - Image interpretation session: 1999. Juxtaglomerular tumor. PMID- 10682785 TI - Image interpretation session: 1999. Bilateral quadriceps tendon rupture and multiple brown tumors in a patient with secondary hyperparathyroidism. PMID- 10682786 TI - Image interpretation session: 1999. Accessory cardiac bronchus. PMID- 10682787 TI - Image interpretation session: 1999. Extramedullary hematopoiesis in a patient with beta thalassemia. PMID- 10682788 TI - Image interpretation session: 1999. Benign mucous membrane pemphigoid (cicatricial pemphigoid). PMID- 10682789 TI - Image interpretation session: 1999. Leiomyosarcoma of the left renal vein. PMID- 10682790 TI - Image interpretation session: 1999. Intraosseous malignant peripheral nerve sheath tumor (malignant schwannoma) in a patient with neurofibromatosis. PMID- 10682791 TI - Image interpretation session: 1999. Inflammatory myoblastic pseudotumor of the trachea (plasma cell granuloma). PMID- 10682792 TI - Image interpretation session: 1999. Desmoplastic infantile ganglioglioma. PMID- 10682793 TI - The visible man: three-dimensional interactive musculoskeletal anatomic atlas of the lower extremity. AB - A personal computer-based interactive musculoskeletal anatomic atlas of the lower extremity has been created by using the Visible Human Male data set. A semiautomatic segmentation program was developed by using an intelligent scissors approach and shape-based interpolation, thus considerably reducing the laborious work of the segmentation and labeling process. Manual contour extractions at 3-mm section intervals and shape-based interpolations of intervening sections of the musculoskeletal structures of the lower extremity were performed. For interactive and realistic three-dimensional display, an efficient binary volume rendering method was developed that introduces the concept of shear-warp factorization and applies a newly developed normal calculation technique. Binary volume rendering reconstructs various structures from a series of two-dimensional sections in a few seconds, thus enabling real-time manipulations of the computerized atlas. All of the muscles, tendons, and bones of the lower extremity have been segmented and labeled. The volume-based three-dimensional interactive atlas supports various interactions including rotation, removal, highlighting with artificial colors, arbitrary cutting operation, transparent view, and descriptive knowledge representation. In addition, browsing through the two-dimensional images of transverse, coronal, and sagittal views with labeling and segmentation information is possible. PMID- 10682794 TI - Potential use of extensible markup language for radiology reporting: a tutorial. AB - One of the main goals of radiology is to communicate imaging information to aid in patient management. Information standards can facilitate communication and help realize this goal. Extensible Markup Language (XML) is a new information transmission standard that was developed to meet the growing need for robust, large-scale World Wide Web applications. XML notation provides a compact document representation scheme that allows radiology reports to be transmitted over the Web as universally understandable, self-defining documents. XML documents can include a report-specific document type definition (DTD) that defines the allowable data fields and values. XML may also be used to generate data entry forms for radiology reporting and help physicians improve the efficiency of the reporting process. XML documents can be used to store reporting results directly, thus allowing pertinent data to be shared on the Web. An XML-based approach can allow users to link information to entities outside the information systems of a given institution. XML-based methods and applications have the potential to promote development of robust radiology reporting systems and integration with broader, enterprise-wide information systems. PMID- 10682795 TI - The effects of corn milling coproducts on growth performance and diet digestibility by beef cattle. AB - Simmental x Angus weanling heifers (n = 96; 239 +/- 2.3 kg) were used in four replications to evaluate three dietary treatments in Trial 1. Treatments were cracked corn-hay diets supplemented with one of three corn milling industry coproducts: dry corn gluten feed (DCGF), dried distillers grains (DDG), and a new modified corn fiber (MCF). In Trial 2, ruminally cannulated mature crossbred beef steers (n = 4; 606 +/-60 kg) were used in a 4 x 4 Latin square with 11-d periods to determine digestibility and ruminal metabolism of limit-fed cracked corn alfalfa haylage diets supplemented with cornstarch (CON), DCGF, DDG, or MCF. During Periods 3 and 4, an in situ study was conducted to compare the rate and extent of CP degradation of DCGF, DDG, and MCF. In Trial 1, there were no differences (P > .10) in initial weights or DM intake. Average daily gain and feed efficiency (G/F) were improved (P < .01) for heifers fed DCGF or DDG vs heifers fed MCF. In Trial 2, no differences (P > . 10) in digestibilities of any nutrients or in ruminal VFA concentrations were observed for steers fed coproducts. The CON supplementation decreased (P < .05) total dietary fiber (TDF) digestibility, improved (P < .10) digestibilities of DM and OM, increased (P < .05) total VFA concentrations and concentrations of propionate and valerate, and decreased (P < .05) concentrations of butyrate, isobutyrate, and isovalerate when compared with the coproducts. Dry corn gluten feed increased (P < .05) and DDG tended (P < .10) to increase percentages of the immediately soluble fraction of CP, and both had increased (P < .05) rates (Kd) and greater (P < .05) extent of ruminal CP degradation than MCF. These data suggest that DCGF and DDG may be utilized in limit-fed high-energy diets without sacrificing performance. Feeding of MCF resulted in poorer performance of heifers, suggesting a limited feeding value that results from high ADIN content and slow in situ protein digestion. PMID- 10682796 TI - Lactation and calf weight traits of mature crossbred cows fed varying daily levels of metabolizable energy. AB - Our objective was to evaluate differences in lactation traits and calf weights produced by F1 cows under varying daily metabolizable energy availability. Measures of milk yields and calf weight traits were recorded on mature F1 cows. The cows were produced from matings of Angus or Hereford dams with sires representing Angus/Hereford, Shorthorn, Galloway, Longhorn, Nellore, and Salers breeds. The cows' daily DM intakes of a diet composed of a corn silage or alfalfa silage plus corn silage were recorded from approximately 2 wk postpartum until the calves were weaned at an average age of 170 d. Milk yield measurements were recorded when the calves were approximately 14, 28, 56, 84, 112, 140, and 168 d of age. Sources of variation considered for the traits of interest included sire breed of the cow (SBC) and the covariates weaning age of the calf and daily metabolizable energy intake (DMEI) of the cow for lactation and calf weights. The linear and quadratic effects were evaluated for DMEI. The SBC x DMEI (linear) interaction was significant for total milk yield. Sire breed of cow differences (P < .05) were observed for milk yield at time of peak yield, persistency, preweaning ADG, and weaning weight. Salers- and Shorthorn-sired cows had greater (P < .05) peak yield than Galloway, Longhorn, or Nellore cross-bred cows but were not significantly different from the Hereford/Angus. Increasing DMEI linearly increased peak yield and total yield (P < .05). Preweaning ADG of calves from Nellore-sired cows was greater (P < .05) than all SBC. Preweaning ADG of calves from Galloway-sired cows was less than all SBC (P < .05). The linear effect of DMEI was heterogeneous across SBC for total yield. The pooled quadratic effect of DMEI was significant for all traits except birth weight. The DMEI for expression of maximum weaning weight was estimated to be 29 Mcal. Feed efficiency ratios for the test period were 28, 27, 30, 25, 28, 32, and 30 g calf weight:Mcal DMEI for reference and 1980s Angus/Hereford-, Shorthorn-, Galloway-, Longhorn-, Nellore-, and Salers-sired cows, respectively, at the DMEI level of 29 Mcal. PMID- 10682797 TI - Twinning rate in Norwegian cattle: frequency, (co)variance components, and genetic trends. AB - The overall twinning rate was shown to increase from .6% in the first parity to 4.0% in the sixth parity, and a positive phenotypic trend for twinning rate was observed during the time period considered (1978 to 1995). The distribution of bulls according to the average percentage of multiple births of daughters in first and second parity showed that some bulls had an extremely low twinning frequency and others exceeded the population mean by approximately six times. (Co)variance components were estimated for twinning in first and second parities using a linear sire model. The analysis included either first- and second-crop daughters (1.4 and .9 million records for first- and second-parity cows, respectively) or first-crop daughters only (.6 and .4 million records for first- and second-parity cows, respectively) from 2,043 sires. Heritability estimates were .7 to .8% in the first parity and 2.8% in the second parity. The genetic correlation between twinning in the first and second parities was approximately 1. Part of the phenotypic trend observed can be explained by a genetic trend for twinning rate. PMID- 10682798 TI - Maternal birth weight breeding value as an additional factor to predict calf birth weight in beef cattle. AB - A total of 1,028 birth weight (BWT) and gestation length (GL) records were collected for calves from 1994 to 1997 in five U.S. Angus herds. Parental BWT EPD and dam BWT maternal breeding values (MBV) computed from the entire U.S. Angus data base after each breeding season were also available. A full model was fit to BWT that contained contemporary group (CG), sire BWT EPD, dam BWT EPD, and dam BWT MBV. A reduced model that dropped dam BWT MBV was also fit. The full model had smaller (P < .01) sum of squares for error than the reduced model. Calf BWT records were then adjusted for CG. Two data sets were formed to include adjusted BWT progeny records from sires with BWT EPD > or = .75 or > or = 1.0 SD above the mean of sires in the data set. Adjusted birth weights were assigned to either > or = .75 or other < .75 SD and > or = 1.0 or < 1.0 SD categories based on the mean BWT of calves in the entire data set. Dams were assigned to either > or =2 .75 or < .75 SD and > or = 1.0 or < 1.0 SD categories based on dam BWT EPD, MBV, or total maternal genetic contribution (TMGC = EPD + MBV). Chi-square analyses showed that dam BWT EPD, MBV, or TMGC categories were not independent (P < .10) of BWT SD categories, indicating that both dam BWT EPD and BWT MBV provide useful information to attenuate calf BWT. Calf BWT records were then adjusted for the overall mean, CG, sire and dam BWT EPD, and dam BWT MBV. Dams were then assigned to five categories: high dam EPD and MBV (HH), high dam EPD and low MBV (HL), low dam EPD and high MBV (LH), low dam EPD and MBV (LL), and other (OTH). High was > or = .75 or > or = 1.0 SD and low was < .75 or < 1.0 SD based on the mean BWT EPD or MBV of dams in the data set. In all adjusted BWT analyses, HH, HL, and LH categories did not differ (P > .05) from each other; however, the LL category was less (P < .05) than all other categories, indicating that calves from LL dams were lighter at birth than expected. For GL, LL was significantly different (P < .05) from only HH. The large differences in birth weight for the LL dams compared to other groups did not seem to be primarily due to shorter GL. Results showed that MBV provided additional information to control BWT; however, when both dam MBV and EPD were low, birth weight of calves was less than expected. Future research should focus on explanations for this interaction. PMID- 10682799 TI - Breed and sex differences in growth curves for two breeds of dog guides. AB - A desirable dog guide weighs 18 to 32 kg as an adult. Male and female German shepherd dogs and male and female Labrador retrievers were weighed between birth and 18 mo of age, with at least one weight recorded after 290 d of age. Growth curves were constructed from 10,484 observations on 880 dogs using the Gompertz function in the form Wt = W(max)exp(-e[-(t-c)/b]), where Wt is weight at time t, Wmax is mature body weight, b is proportional to duration of growth, c is age at point of inflection, and t is age in days. Estimates for mature body weight were 2.4 +/- .3 kg higher for Labrador retrievers than for German shepherd dogs and 4.7 +/- .2 kg higher for males than for females. Male Labrador retrievers were closest to the upper limit for desirable weight, with an average estimated mature weight of 31.4 +/- .3 kg. Duration of growth, 4b + c, was not different between the breeds; however, the estimate for males was 8 +/- 5d longer than for females. Female Labrador retrievers had the shortest estimate for growth of 319 +/- 6 d. The estimate for age at the point of inflection was 3.6 +/- 1.2 d greater for males than for females, but not different between breeds. A better understanding of growth curves for dog guides may aid in estimating mature weight at a young age, thus allowing earlier breeding and training decisions to be made and increasing genetic change per year. PMID- 10682800 TI - Variance and covariance components for weaning weight for Herefords in three countries. AB - Records from the Hereford Associations of the United States (USA), Canada, and Uruguay were used to estimate genetic and phenotypic variances and covariances for weaning weight. Estimation was done using a complete animal model, relatively large data sets, and the same methodology for the three countries in order to determine whether genetic parameters for weaning weight were homogeneous across environments. Data were composed of 2,322,722, 487,661, and 102,986 edited weaning weight records for USA, Canada, and Uruguay, respectively. Ten samples were obtained from each country by eliminating data from small herds with fewer than 500 records, selecting herds at random from the entire data set after removing the small herds, and then retaining the direct-sire-connected contemporary groups within each sample. The final sample sizes ranged from 9,832 to 46,377 records. An accelerated EM-REML algorithm was used in estimating the (co)variance components in each sample. The estimates were pooled by calculating the arithmetic mean of the 10 samples from within each country. Direct and maternal (in parentheses) heritability estimates were .24 (.16), .20 (.16), and .23 (.18) for USA, Canada, and Uruguay, respectively. Maternal heritabilities reported here are nearly 50% smaller than the values currently used in national genetic evaluation for the breed, which were estimated using sire-maternal grandsire models. Covariance between direct and maternal was negative in all countries, accounting for 6, 8, and 10% of the total phenotypic variation, and the total dam effect was 32.5, 37.0, and 34.0% in USA, Canada, and Uruguay, respectively. Total heritabilities were similar among the countries, with values of .19, .19, and .17 for the three respective countries. The similarity of genetic and environmental parameters across the three countries suggests that joint genetic evaluation is feasible across environments provided that the genotype x environment interaction is negligible and can be ignored. PMID- 10682801 TI - Few differences found between early- and late-weaned pigs raised in the same environment. AB - Segregation and medicated early weaning are technologies used to optimize the productivity and health of pigs, but these practices may also cause aberrant behaviors indicative of stress. Thus, differences in early- (=10 d of age) and late- (=30 d of age) weaned pigs were investigated. At weaning, pigs were housed in groups of four in 16 pens (eight pens per treatment) in the same facility, and, thus, they were not segregated. Body weights were recorded at birth, weaning, and at approximately 42, 65, 102, 137, and 165 d of age (at slaughter). One-minute, instantaneous scan samples during a 10-min period (at 0600, 1000, 1400, and 1800) were used to record the frequency of lying, standing, and sitting, total number of drinks, feeder investigations, and time spent playing/fighting on 2, 3, and 4 d after weaning. Five-minute, direct observations of each pig were conducted at approximately 40, 60, 80, and 150 d of age. Direct observations were also made of the entire pen for 10 min at approximately 50, 95, 123, and 160 d of age to record aberrant behaviors. At 62 d of age, a handling and blood collection stress was imposed. At 165 d of age, a second stress test was conducted in response to rough handling and transport. Early-weaned pigs spent more time playing/ fighting (P < .006) than late-weaned pigs during the 4 d after weaning, manipulated conspecifics more often at 40 d of age (P < .002), had greater percentage of hemoglobin (P < .03) during Stress Test 1, had greater ADG at 42 d of age (P < .03), and had greater hypothalamic growth hormone-releasing hormone receptor mRNA at slaughter (P < .06). Late-weaned pigs had greater ADG between 137 and 165 d of age (P < .03) and greater pro-opiomelanocortin at slaughter (P < .04). Overall, most differences found between early-weaned and late-weaned pigs were evident soon after weaning, but they disappeared before slaughter. PMID- 10682802 TI - Effects of birth weight and postnatal nutrition on neonatal sheep: II. Skeletal muscle growth and development. AB - This study investigated effects of birth weight and postnatal nutrition on growth and development of skeletal muscles in neonatal lambs. Low (L; mean +/- SD 2.289 +/- .341 kg, n = 28) and high (H; 4.840 +/- .446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG 337 g, n = 20) or slowly (ADG 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, weight of semitendinosus (ST) muscle in L lambs was 43% that in H lambs; aggregate weights of ST and seven other dissected muscles were similarly reduced. In ST muscle of L lambs, mass of DNA, RNA, and protein were also significantly reduced to levels 67, 60, and 34%, respectively, of those in H lambs. However, myofiber numbers of ST, tibialis caudalis, or soleus muscles did not differ between the L and H birth weight lambs and did not change during postnatal growth. During postnatal rearing, daily accretion rate of dissected muscle was lower in L than in H lambs. Accretion of muscle per kilogram of gain in empty body weight (EBW) was reduced in the slowly grown L lambs compared with their H counterparts, although the difference was less pronounced between the rapidly grown L and H lambs. Throughout the postnatal growth period, ST muscle of L lambs contained less DNA with a higher protein:DNA ratio at any given muscle weight than that of H lambs. Slowly grown lambs had heavier muscles at any given EBW than rapidly grown lambs. Content of DNA and protein:DNA ratio in ST muscle were unaffected by postnatal nutrition, but RNA content and RNA:DNA were greater and protein:RNA was lower at any given muscle weight in rapidly grown lambs. Results suggest that myofiber number in fetal sheep muscles is established before the presumed, negative effects of inadequate fetal nutrient supply on skeletal muscle growth and development become apparent. However, proliferation of myonuclei may be influenced by fetal nutrition in late pregnancy. Reduced myonuclei number in severely growth-retarded newborn lambs may limit the capacity for postnatal growth of skeletal muscles. PMID- 10682803 TI - Ovine adipose tissue monounsaturated fat content is correlated to depot-specific expression of the stearoyl-CoA desaturase gene. AB - The basis for the variation in fatty acid composition in different ovine adipose tissue depots was investigated. The proportion of stearic (C18:0) and oleic (C18:1) acids vary in a site-specific fashion; abdominal depots (omental and perirenal) contain relatively more C18:0 than C18:1, and carcass depots, especially sternum, have a markedly higher proportion of C18:1. Additionally, expression of a number of lipogenic enzyme genes (stearoyl-CoA desaturase [SCD], acetyl-CoA carboxylase-alpha [ACC-alpha], lipoprotein lipase [LPL]) and the cytoskeletal protein gene alpha-tubulin vary among depots, although the pattern of variation differs for each mRNA. When these expression data were related to the mean cell volume of adipocytes pooled from all depots, a significant pattern emerged: expression of the ACC-alpha, LPL, and alpha-tubulin genes was highly correlated with the size of adipocytes. In contrast, when the expression of SCD mRNA was assessed as a function of mean cell volume, two populations of adipocytes emerged: no significant correlation was found between the expression of SCD mRNA per adipocyte and mean cell volume for the abdominal depots, although a highly significant correlation was observed between SCD gene expression and mean cell volume for the carcass and epicardial depots. Similarly, a highly significant correlation was found for the amount of C18:1 per adipocyte and the abundance of SCD mRNA per adipocyte for the carcass and epicardial depots, whereas no significant correlation was observed for these traits for the omental and perirenal depots. Thus, the SCD gene seems to be regulated in a depot specific fashion and in a manner distinct from that of the ACC and LPL genes. PMID- 10682804 TI - Messenger ribonucleic acid expression of the MyoD gene family in muscle tissue at slaughter in relation to selection for porcine growth rate. AB - Livestock meat production capacity is related to muscle fiber numbers and growth. Muscle fibers develop during early embryonic development from proliferating and differentiating myoblasts. Post-natal muscle growth requires satellite cell proliferation and differentiation. Myoblast and satellite cell proliferation and differentiation is regulated by the genes of the MyoD gene family (myogenin, myf 5, myf-6, and MyoD1). Our aim was to study the mRNA expression of these genes in postnatal muscle tissue in relation to porcine selection for growth rate or leanness. Five boars from a line selected for fast growth (F-line) and five boars from a line selected against backfat thickness (L-line) were slaughtered, and biopsies were taken from 12 muscles. Between-line effects, within-line effects in relation to the performance of the pigs, and muscle-specific effects were studied. Comparing the F-line with the L-line revealed significantly greater myogenin, myf-5, and MyoD1 mRNA expression in some muscles of the F-line. The expression of myf-6 showed a tendency for the opposite effect in some muscles. Muscles were ordered by their muscle-specific growth rate (b-value). Within-line evaluation of the data revealed a systematic muscle effect for the myf-6 expression level in the F-line because higher b-values correlated with increased myf-6 expression level. Backfat thickness was negatively related to myogenin expression in the F-line. A relationship was found between myogenin:MyoD1 mRNA expression ratio and meat color/muscle fiber type composition in the L-line. Furthermore, the myogenin:MyoD1 ratio was greater in muscles from F-line boars than in muscles from L-line boars, which relates to the difference between the lines in muscle fiber type. We conclude that the mRNA levels of the MyoD genes in porcine muscle tissue at slaughter showed different relationships to selection for growth rate when evaluated between selection lines and within selection lines. PMID- 10682805 TI - Protein kinetics in callipyge lambs. AB - The objectives for this experiment were to determine the effect of the callipyge phenotype on protein kinetics. We studied callipyge and normal lambs (n = 37) at 5, 8, and 11 wk of age (n = 4 to 7/ group) to determine how protein kinetics are altered by this trait. Total protein, DNA, and RNA and calpastatin activity were measured in five skeletal muscles and in the heart, kidneys, and liver, and protein accretion rates were calculated. At 8 wk, the fractional synthesis rates of proteins in these tissues were measured in vivo using a primed, continuous 8-h infusion of [2H5]phenylalanine. Fractional rates of protein degradation were estimated by differences. At 5 wk of age, muscle weights, protein mass, protein:DNA, RNA:DNA, and calpastatin activity were higher (P < .05) for callipyge, and protein mass differences continued to increase through 11 wk. At 8 wk, fractional rates of protein synthesis and degradation were lower (P < .05) in callipyge than in normal lambs. The organs of callipyge lambs exhibited reduced growth at 11 wk. Thus, enhanced muscle growth seems to be maintained in callipyge lambs by reduced protein degradation rather than increased protein synthesis. However, we cannot exclude the possibility that the initial onset of the callipyge condition may be caused by an increase in the fractional rate of protein synthesis. PMID- 10682806 TI - Added dietary pyridoxine, but not thiamin, improves weanling pig growth performance. AB - We conducted two trials to determine the effects of added dietary pyridoxine (vitamin B6) or thiamin (vitamin B1) on growth performance of weanling pigs. In Exp. 1, weanling pigs (n = 180, initially 5.55 +/- .84 kg, and 21 +/- 2 d of age) were fed either a control diet (no added pyridoxine or thiamin) or the control diet with added thiamin (2.8 or 5.5 mg/kg) from thiamin mononitrate or pyridoxine (3.9 or 7.7 mg/kg) from pyridoxine HC1. These five diets were fed in meal form in two phases (d0 to 14 and 14 to 35 after weaning), with identical vitamin concentrations in both phases. From d 0 to 14 after weaning, pigs fed added pyridoxine had increased (quadratic, P < .05) ADG and ADFI; pigs fed 3.9 mg/kg of added pyridoxine had the greatest improvement. From d 14 to 35 and 0 to 35, ADG and ADFI increased (linear P = .06) for pigs fed increasing pyridoxine. Growth performance was not improved by added thiamin. In Exp. 2, weanling pigs (n = 216, initially 6.08 +/- 1.13 kg, and 21 +/- 2 d of age) were fed a control diet or the control diet with 1.1, 2.2, 3.3, 4.4, or 5.5 mg/kg of added pyridoxine from pyridoxine HCl. From d 0 to 14 after weaning, increasing pyridoxine increased (quadratic, P < .05) ADG and ADFI; pigs fed 3.3 mg/kg of added pyridoxine had the greatest ADG and ADFI. Break-point analysis suggested a requirement estimate of 3.3 and 3.0 mg/kg of added pyridoxine to maximize ADG and ADFI, respectively. From d 14 to 35 or 0 to 35, increasing pyridoxine had no effect (P > .10) on pig growth performance. These results suggest that adding 3.3 mg/kg of pyridoxine (7.1 to 7.9 mg/kg of total pyridoxine) to diets fed from d 0 to 14 after weaning can improve pig growth performance. PMID- 10682807 TI - Efficacy of yeast phytase in improving phosphorus bioavailability in a corn soybean meal-based diet for growing pigs. AB - Crossbred barrows (n = 66; 6 wk old) were used in a 6-wk experiment to evaluate the efficacy of phytase from yeast or Aspergillus niger on performance, tibial characteristics, and serum inorganic P concentration. We also investigated the stability of these phytases in acidic solutions with pepsin, which simulated gastric conditions. Pigs were fed a P-adequate diet containing .34% nonphytate-P or a low-P diet containing .20% nonphytate-P. The low-P diet was supplemented with 0, 1,000, 2,000, or 4,000 phytase units (PU; the activity at optimal pH, i.e., pH 4.2 for yeast phytase and pH 5.5 for phytase from Aspergillus niger)/kg of yeast phytase, or 1,000 PU/kg phytase from Aspergillus niger. The graded level of yeast phytase linearly increased ADG (P = .047), tibial weight (P = .091), tibial density (P < .001), and P concentration in tibial cortex (P = .018). Aspergillus niger phytase also increased ADG (P = .022), serum inorganic P concentration (P < .001), tibial density (P = .007), and tibial P concentration (P = .025). The pigs given 1,000 PU/kg Aspergillus niger phytase showed greater ADG (P = .091), tibial density (P= .001), and tibial P concentration (P = .062) than those given 1,000 PU/kg yeast phytase. No measurements differed (P > .31) between the pigs given 1,000 PU/kg Aspergillus niger phytase and those given 4,000 PU/kg yeast phytase. These results suggested that yeast phytase improves bioavailability of P in the diet for growing pigs but the efficacy of yeast phytase is less than that of Aspergillus niger phytase. During incubation in acidic solutions with pepsin, yeast phytase (P < .001) lost more of its activity than Aspergillus niger phytase. This lesser stability of yeast phytase may be responsible for the poorer efficacy of yeast phytase than that of Aspergillus niger. In summary, supplementation of swine diets with yeast phytase is beneficial, but its efficacy is less than that of Aspergillus niger phytase. PMID- 10682808 TI - Effect of organic and inorganic selenium sources and levels on sow colostrum and milk selenium content. AB - A study was conducted to evaluate the short-term effects of feeding two dietary Se sources at various Se levels on the transfer of Se to the dam's milk and nursing pig. Six dietary treatments were arranged in a 2 x 2 factorial arrangement with two additional treatments in a randomized complete block designed experiment. Inorganic (sodium selenite) or organic (Se-enriched yeast) Se sources were added to the diet at .15 or .30 ppm Se. A non-Se-fortified corn soybean meal basal diet served as a negative control, and a sixth group was fed .15 ppm Se from both inorganic and organic Se sources. A total of 43 sows were fed their treatment diets at 2.2 kg/d from 6 d prepartum to parturition and at full feed through a 14-d lactation period. Ten sows were initially bled at 6 d prepartum, and three sows and three pigs from their litters were bled at 7 and 14 d postpartum. Serum was analyzed for its Se concentration and glutathione peroxidase (GSH-Px) activity. Colostrum was collected within 12 h postpartum and milk at 7 and 14 d of lactation. When the basal diet was fed, sow serum GSH-Px activity declined from 6 d prepartum and remained low throughout lactation. When dietary Se levels increased, sow serum Se concentration and serum GSH-Px activity increased (P < .05) at both 7 and 14 d postpartum. The short-term feeding of either Se source at .15 or .30 ppm Se did not affect colostrum Se content when inorganic Se was fed, but it was increased when organic Se was provided. This resulted in a significant Se source x Se level interaction (P < .01). Milk Se at 7 and 14 d postpartum was 2.5 to 3 times higher when the organic Se source was provided and resulted in a significant Se source x Se level interaction (P < .05). When the combination of inorganic and organic Se was fed at .15 ppm Se, colostrum and milk Se contents were similar to those of sows fed .15 ppm Se from the organic Se source. Pig serum GSH-Px activity was not affected at 7 and 14 d of age by dietary Se level or Se source fed to the sow, but serum Se increased (P < .05) as dietary Se level increased, particularly when sows had been fed organic Se. The results demonstrated that organic Se increased milk Se content more than did inorganic Se and increased the nursing pig's serum Se. These results indicate that inorganic Se was more biologically available for sow serum GSH-Px activity, but organic Se was more effectively incorporated into milk. PMID- 10682809 TI - Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs. AB - Thirty-two crossbred barrows were used to investigate the effects of dietary Ca:total P (tP) ratios in phytase-supplemented diets on the apparent absorption of P and Ca in the small intestine, cecum, and colon. Three Ca:tP ratio treatments (1.5:1, 1.3:1, or 1.0:1) were created by adjusting the amount of ground limestone added to the basal low-P grower (.39% tP including .07% added inorganic P) and finisher (.32% tP without added inorganic P) diets. All low-P ratio diets were supplemented with Natuphos phytase at 500 units/kg. A positive control diet without phytase supplementation contained adequate P and Ca to meet dietary requirements. At 123 kg, the pigs were slaughtered and the contents of ileum, cecum, and colon were collected. Lowering the dietary Ca:tP ratio in the diets containing phytase linearly increased (P < .01) the apparent absorption (% and g/d) of P in the small intestine, but Ca absorption was not affected. Pigs fed the low-P diet with a Ca:tP ratio of 1.0:1 had an apparent absorption (g/d) of P or Ca similar to that of pigs fed the control diet, which was adequate in Ca and P. Averaged across all diets, the apparent absorption of P was highest when measured at the cecum, and the apparent absorption of Ca was highest when measured at the colon. In conclusion, lowering the dietary Ca:tP ratio to 1.0:1 in a low-P diet containing phytase increased the apparent absorption of P in the small intestine. Furthermore, a significant amount of P was absorbed in the cecum. PMID- 10682810 TI - Effect of vitamin E sources (RRR- or all-rac-alpha-tocopheryl acetate) and levels on sow reproductive performance, serum, tissue, and milk alpha-tocopherol contents over a five-parity period, and the effects on the progeny. AB - Two dietary sources of vitamin E (DL-alpha-tocopheryl acetate [DL-beta-TAc], or D alpha-tocopheryl acetate [DL-alpha-TAc]) at two dietary supplemental levels (30 vs 60 IU/kg) were evaluated in reproducing sows over a five-parity period. The experiment was a 2 x 2 factorial arrangement of treatments conducted as a randomized complete block in two replicates. A total of 48 gilts were fed their treatment diets from 40 kg BW through five parities, reflecting a total of 171 farrowings. Reproductive measurements of litter size, sow weight, and back-fat thickness were collected. The incidence of mastitis-metritis-agalactia (MMA) and fluid discharge from the vagina were evaluated for each sow on each of the first 3 d postpartum. Sows were bled at periodic intervals during gestation and at weaning (21 d) and serum was frozen. After the fifth parity, two to four sows from each treatment group were killed and tissues collected. At birth, two to three neonatal pigs were killed from each sow treatment group within each parity and livers were collected and frozen. In addition, three pigs from each litter from three to four sows per treatment group within each parity were bled at weaning and serum was saved. Six pigs from each sow group at weaning of Parity 5 were also killed and livers were collected and frozen. Sow and pig sera and tissues were analyzed for a-tocopherol. There was no effect (P > .15) of vitamin E source or level on the various sow reproductive measurements, litter size, or the incidences of MMA or fluid discharges from the vagina. Feeding D-alpha-TAc compared with DL-alpha-TAc or 60 IU compared with 30 IU vitamin E/kg diet resulted in higher (P < .01) sow serum, colostrum, and milk alpha-tocopherol contents at each measurement period. Sow liver, adipose, lung, and heart alpha tocopherol contents were also higher (P < .01) when the 60 IU vitamin E level had been fed. Both serum and liver a-tocopherol contents in 21-d-old nursing pigs were higher (P < .01) when the sow had been fed D-alpha-TAc compared with the DL alpha-TAc source or when the 60 IU level had been fed. There were no vitamin E source x vitamin E level interactions (P > .15) for the various alpha-tocopherol measurements. Although the supplemental vitamin E sources were provided on an equivalent IU basis, these results suggest that D-alpha-TAc has a higher equivalency than DL-alpha-TAc on an IU basis, but higher dietary levels also resulted in higher sow and pig alpha-tocopherol contents. PMID- 10682811 TI - Effect of feather meal as a source of valine for lactating sows. AB - An experiment was conducted to evaluate feather meal as a source of Val in lactating sow diets. Sows (five farrowing groups; mean parity = 2.34) were allotted to one of two dietary treatments on the basis of ancestry, parity, and weight and date of d 110 of gestation. The treatment diets included 1) corn soybean meal lactation diet (n = 40) or 2) corn-soybean meal lactation diet with 2.5% feather meal (n = 39). The diets were formulated on an equal Lys basis. All litters were adjusted to 10 pigs within 24 h after farrowing, and all sows weaned at least nine pigs. Sows were bled at 110 d of gestation and at weaning, and serum urea N was determined. Backfat thickness was determined ultrasonically at 110 d of gestation and at weaning. Serum urea N and backfat thickness at d 110 of gestation were used as covariates for serum urea N and backfat thickness at weaning, respectively. The litter response criteria (weaning weight, litter weight gain, and percentage survival) were not affected (P > .10) by feather meal. The sow response criteria (weaning weight, weight loss per day, weaning backfat thickness, change in backfat thickness, ADFI, and days to estrus) were not affected (P > .10) by feather meal. Sows fed feather meal had increased (P < .01) serum urea N and tended (P = .15) to have decreased sow weaning weight. Following the initial analysis of the data, the data set was split into two groups: 1) sows with litters gaining less than 2.17 kg/d (n = 19 and 20 for control and feather meal diets, respectively) and 2) sows with litters gaining more than 2.17 kg/d (n = 21 and 19 for control and feather meal diets, respectively). These two groups were analyzed separately. In sows with litters gaining less than 2.17 kg/d, the litter and sow criteria were not affected (P > .10) by treatment. In sows with litters gaining more than 2.17 kg/d, sow weaning weight was decreased (P < .04) and sow weight loss (P < .02) and serum urea N (P < .01) were increased in sows fed feather meal. Feather meal (as a source of Val) did not improve litter weight gain, but it increased serum urea N. PMID- 10682812 TI - Physiological responses of Brahman and Hereford steers to an acute ergotamine challenge. AB - Research was conducted to evaluate the sensitivity of Bos indicus and Bos taurus cattle to a tall fescue ergopeptine alkaloid by assessing vital sign responses. Eight Polled Hereford and seven Red Brahman steers received bolus i.v. injections of ergotamine tartrate and saline vehicle in a simple cross-over design. Physiological traits measured 30 min and immediately before and 30, 60, and 90 min after treatment were respiration rate, rectal temperature, skin temperature at the tailhead and tail tip, systolic and diastolic blood pressure, and heart rate. Blood samples were collected immediately before and 105 min after treatments to determine plasma prolactin and cortisol concentrations. Steers were fed a fescue-free diet in drylot. Ambient temperature and relative humidity averaged 31 degrees C and 55%, respectively, during data collection. No breed x treatment x time interactions were apparent for vital signs. The treatment x time interaction was significant (P < .05) for blood pressure and skin temperature. Ergotamine increased (P < .01) blood pressure and decreased (P < .01) skin temperature. The breed x treatment x time interactions were significant for prolactin (P < .1) and cortisol (P < .01). Ergotamine decreased plasma (P < .01) prolactin and increased (P < .01) cortisol concentrations in both breeds, despite some breed variation. Across all traits, Brahman and Hereford steers responded similarly to acute ergotamine exposure, indicating that the breeds are alike in acute sensitivity to a systemically administered ergopeptine alkaloid associated with fescue toxicosis. PMID- 10682813 TI - Morphology of luminal and glandular epithelial cells from pig endometrium grown on plastic or extracellular matrices. AB - Luminal (LE) and glandular epithelial (GE) cells from d-13 pregnant pigs were cultured on plastic, matrix secreted from endometrial stroma, and EHS matrix (Matrigel) in culture medium (RPMI-1640) supplemented with 20 and 10% fetal bovine serum, respectively. After culture for 7 and 14 d, GE and LE cells were prepared for transmission and GE cells for scanning electron microscopy. The two types of endometrial epithelial cells displayed different morphological characteristics when grown on different culture substrates. On plastic, the GE and LE cells formed flattened monolayers. However, stroma-secreted matrix directed the polarization of endometrial epithelia. The GE and LE cells reacted differently to thick Matrigel coatings; LE cells formed a colony after 7 d of culture and then proliferated further to form a colony with a cavity, but GE cells organized to form a colony with a shallow depression in the center at 7 d and developed duct-like structures after 14 d in vitro. Luminal epithelial cells grown on either diluted or thin-coated Matrigel and grown on stroma-secreted matrix formed a monolayer but no three-dimensional structures. PMID- 10682814 TI - Plasma fatty acids, prostaglandin F2alpha metabolite, and reproductive response in postpartum heifers fed rumen bypass fat. AB - An experiment was conducted to determine whether feeding rumen-protected fatty acids (FA) to postpartum heifers would increase plasma concentrations of linoleic acid and PGF2, metabolite (PGFM), shorten the interval from calving to first increase in plasma concentrations of progesterone (P4), and increase pregnancy rate relative to controls. Hereford x Angus heifers (346 kg) were assigned randomly to treatments containing either lipid or barley supplemented diets for the first 30 d postpartum. Lipid was .23 kg.heifer(-1).d(-1) of calcium salts of FA (CSFA; n = 20), and an isocaloric amount of barley served as the control (n = 19). Supplements, with .23 kg of barley as a vehicle, and a basal diet of meadow and alfalfa hays were pen fed to heifers (5/pen). Heifers were bled on alternate days (d1 to 30) and twice weekly (d 30 to 2 wk after first estrus) for RIA of plasma PGFM and P4, respectively. Weight percentage of major FA in plasma on d1 and 7 was determined with gas chromatography. First behavioral estrus was detected by use of intact bulls and confirmed by an increase in plasma P4. On d 7, but not d 1, plasma from heifers fed CSFA had altered proportions of major FA (P < .01), including an increase in linoleic acid compared with those of controls (29.1 vs 25.6% of total FA; SE = .75; P < .01). Analysis of variance of contrast variables revealed an effect of treatment on direction of change in PGFM from d 3 to 5 (P < .01). By d 7 and on d 9, plasma concentrations of PGFM were greater in heifers fed CSFA than in controls (P = .02 and P = .06, respectively). There was no difference in plasma concentration of PGFM between treatments on d 1, 3, 5, 11, 13, and 15 postpartum (P = .80, .17, .52, .82, .46, and .77, respectively). Days to first estrus with ovulation, pregnancy rate, and calving interval were not affected by treatments (P = .58, .52, and .24, respectively). Although supplemental lipid fed to primiparous beef heifers increased plasma levels of linoleic acid and production of PGFM in the early postpartum period, it did not improve the fertility of these heifers in the subsequent breeding season. PMID- 10682815 TI - Use of a small dose of estradiol benzoate during diestrus to synchronize development of the ovulatory follicle in cattle. AB - We tested the hypothesis that a small dose of estradiol benzoate (EB) at the midstage of the estrous cycle in cattle would synchronize the subsequent pattern of ovarian follicular development, estrus, and ovulation. Nonlactating Friesian cows received either 1 mg of EB i.m. on d 13 of the estrous cycle (T; n = 12; estrus = d0) or served as untreated controls (C; n = 12). Their ovaries were examined daily with transrectal ultrasonography from d 7, and blood samples were collected 0, 2, 4, 8, 24, and 48 h after treatment on d 13. Plasma concentrations of estradiol-17beta were elevated to 12 pg/mL during the initial 24 h following treatment, compared with a baseline of 1 pg/mL in untreated controls (P < .001). Progesterone concentrations in cows of the T group declined between 24 and 48 h after treatment (-3.2 +/- .5 ng/mL) compared with little change in concentrations of progesterone in cows of the C group at this time (P < .01). This difference was coincident with an earlier time to regression of the corpus luteum in cows of the T group. Disregarding treatment groups, the second dominant follicle of the estrous cycle (DF2) emerged on d 10.6 +/- .3 and was 9.4 +/- .4 mm in diameter on d 13. Further growth of the DF2 was halted by EB treatment on d 13. Cessation of growth occurred irrespective of whether the DF2 was in the early or late growth phase, and a new follicular wave emerged 4.5 +/- .2 d later. The dominant follicle from this wave (DF3) ovulated 5 d after emergence in most cases. During the estrous cycle of every cow in the T group, there were three waves of follicular development (3-wave), whereas the ratio of 2:3 waves of follicular development in cows of the C group was 1:3. Consequently, the interval from emergence to ovulation of the ovulatory dominant follicle in cows of the C group ranged from 3 to 11 d. The dynamics of ovarian follicular wave development during the estrous cycle can be strategically manipulated by treating with a small dose of EB to synchronize proestrous development of the ovulatory follicle. PMID- 10682816 TI - Reduced oxygen tension and EDTA improve bovine zygote development in a chemically defined medium. AB - Bovine zygotes produced by in vitro oocyte maturation and fertilization were cultured for 7.5 d in a chemically defined medium without serum or proteins, except .12 IU/mL of insulin. In Exp. 1, embryos were cultured in approximately 20% oxygen (i.e., 5% CO2 in air) or 5% CO2; 5% O2; 90% N2, with the metal chelators EDTA or diethylenetetraaminopentaacetic acid (DTPA) at 0, 5, 25, or 125 microM. More (P < .01) embryos developed to blastocysts at 5% O2 (17%) than at 20% O2 (7%). Also, embryos grown at 5% O2 averaged more cells than embryos cultured at -20% O2 (38 vs 29 cells for morulae and blastocysts and 15 vs 12 cells including all embryos; P < .05). There were interactions (P < .01) among chelator, concentration of chelator, and oxygen tension. The most efficacious treatments were 5 microM EDTA at 5 or -20% O2 (24 and 20% blastocysts), 5 microM DTPA at 5% O2 (28% blastocysts), and 25 microM EDTA at 5% O2 (25% blastocysts). High concentrations of either chelator were detrimental, especially at -20% O2. In Exp. 2, a smaller range of chelator concentrations was compared (EDTA: 3, 9, 27, or 81 microM, DTPA: 3 or 15 microM) in 5% O2. More embryos developed to blastocysts and expanded blastocysts with 3 microM EDTA than the control without a chelator (20 and 16% vs 7 and 3%, respectively; P < .05). However, in Exp. 3, which concerned embryo development in .33, 1, 3, or 27 microM EDTA and .33, 1, or 3 microM DTPA, no concentration of either chelator was better (P > . 1) than the control. PMID- 10682817 TI - Amino acid flux in ruminal and gastric veins of sheep: effects of ruminal and omasal injections of free amino acids and carnosine. AB - The possibility of free amino acid (FAA) and peptide absorption across the ruminant stomach wall was studied in multicatheterized wethers fed every 12 h. During the last third of the feeding cycle, two intraruminal or intraomasal injections of solutions containing increasing amounts of Ser, Gly, Val, Met, Phe, Lys, and carnosine were successively performed. Before injections, a net uptake of each of these FAA was measured in the ruminal and the gastric veins. The ruminal injections produced a linear increase in ruminal FAA concentration. The highest ruminal concentrations (observed with 3 g of FAA and carnosine) ranged between 5 and 14 mM. After ruminal injections, Ser (P < .05), Gly (P < .05), Val (P < .05), Met (P < .10), and Lys (P < .10) uptake decreased and carnosine net release linearly increased (P < .05), suggesting absorption across the ruminal epithelium. Owing to the low net flux generated by high ruminal concentration, the ruminal epithelium permeability to these molecules seemed to be low. After omasal injections, net flux of injected FAA were not modified, suggesting a low permeability of the gastric epithelia to FAA. Carnosine net release linearily increased (P < .05) with increasing level of carnosine injection, indicating the possibility of dipeptide absorption at the gastric level. This study demonstrated in vivo that the stomach epithelia possess the capacity to absorb FAA and small peptides; however, the permeability of these epithelia to these molecules seemed limited. PMID- 10682818 TI - Effects of base ingredient in cooked molasses blocks on intake and digestion of prairie hay by beef steers. AB - Twelve steers (332 kg) were used in three simultaneous 4 x 3 incomplete Latin squares to evaluate effects of beet molasses (BEET), cane molasses (CANE), or concentrated separator by-product (CSB) as base ingredients in cooked molasses blocks on intake and digestion of prairie hay and ruminal characteristics. All steers had ad libitum access to prairie hay (5.9% CP and 69.4% NDF; DM basis). The four experimental treatments included a control (no supplement) and three cooked molasses blocks, based on BEET, CANE, or CSB, fed daily at .125% of BW (.42 kg/d as-fed, .13 kg/d CP). Forage OM, NDF, and N intakes; digestible OM, NDF, and N intakes; and total tract OM and N digestibilities (percentage of intake) were greater (P < .05) for steers fed cooked molasses blocks than for control steers. Total tract OM digestibility was greater (P < or = .06) for steers fed BEET blocks (54.0%) than for those fed CSB (52.1%) or CANE blocks (52.2%). Digestion of NDF was greatest (P < .05) for steers fed BEET blocks (51.9%) and tended to be greater (P < .07) for steers fed CANE (49.3%) or CSB blocks (49.3%) than for control steers (46.9%). Ruminal ammonia concentrations were greater (P < .05) for steers fed cooked molasses blocks (.89 mM) than for control steers (.21 mM); this was primarily due to increases to 4.6 mM at 2 h postfeeding for steers fed blocks. Concentrations of total VFA in ruminal fluid were greater (P < .05) for steers fed BEET (92.7 mM) and CSB (88.1 mM) blocks than for control steers (80.3 mM), whereas concentrations for steers fed CANE blocks were intermediate (85.4 mM). Steers supplemented with cooked molasses blocks had greater molar percentages of butyrate than did control steers, particularly shortly after feeding. In summary, supplementation with cooked molasses blocks increased forage intake and digestion. The three base ingredients elicited similar responses, although steers fed BEET had slightly greater OM and NDF digestibilities than those fed CANE or CSB. PMID- 10682819 TI - Efficacy of laidlomycin propionate in low-protein diets fed to growing beef steers: effects on steer performance and ruminal nitrogen metabolism. AB - We conducted two experiments to evaluate the effect of the ionophore laidlomycin propionate (LP) on steer performance and ruminal N metabolism. Experiment 1 was a 91-d growth study evaluating the growth and ruminal characteristics of steer calves consuming supplemental LP. Steers (n = 96; 255 +/- 3 kg; four steers/pen; six pens/treatment) were used in a randomized complete block design with a 2 x 2 factorial arrangement of treatments consisting of two levels of dietary CP (formulated to be 10.5 and 12.5% of DM) with and without LP (11 mg/kg diet DM). Ruminal fluid was collected via stomach tube on d 91 from one steer randomly selected from each pen. No CP x LP interactions were observed with performance data (P > .64). Final weight and total gain were greater (P < .07) for 12.5% CP and LP compared with 10.5% CP and control steers, respectively. Also, DMI was increased (P = .08) with 12.5% CP but not with LP supplementation (P = .36). In addition, ADG and gain:feed ratio were greater (P < .03) for both 12.5% CP and supplemental LP. Ruminal NH3 N concentration was greater (P < .09) with 12.5% CP and LP. Total VFA concentration and molar proportion of acetate were not affected by treatment (P > .11). However, propionate concentration was increased (P < .09) with 12.5% CP and LP, and acetate:propionate was lower (P = .02) with LP supplementation. In Exp. 2, six steers were used in a replicated 3 x 3 Latin square design to compare ruminal fermentation and protein degradation in steers without ionophore feeding or adapted to LP or monensin. In vitro deamination of amino acids by adapted ruminal microbes was also assessed. Ionophore supplementation decreased (P = .07) ruminal NH3 N concentration compared with control steers, and LP increased (P = .02) ruminal NH3 N compared with monensin. Molar proportion of acetate was decreased (P = .02) and propionate increased (P = .01) with ionophore treatment. Consequently, ionophore supplementation depressed the acetate:propionate ratio (P = .01). In situ degradation rate of soybean meal (SBM) CP was greater (P = .09) with ionophore treatment, but estimates of SBM undegradable intake protein were not altered by treatment (P > .25). Microbial specific activity of net NH3 N release and alpha-amino N degradation were decreased (P < .04) with ionophores. Based on this study, LP and monensin did not affect the extent of ruminal degradation of SBM CP but decreased amino acid deamination. PMID- 10682820 TI - Influence of the novel urease inhibitor N-(n-butyl) thiophosphoric triamide on ruminant nitrogen metabolism: I. In vitro urea kinetics and substrate digestion. AB - Two in vitro digestion experiments were conducted to evaluate the influence of the novel urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) on in vitro urea kinetics, substrate digestion, and fermentation characteristics. In Exp. 1, in vitro incubations were conducted in 50-mL test tubes containing .25 g of ground fescue hay to which 0, 6.5, 13, 26, or 52 mg of NBPT in a buffered ruminal fluid innoculum was added. Tubes were incubated in triplicate at 39 degrees C and replicated on consecutive days, with NH3 N and urea concentrations measured at 0, 10, 30, 60, 120, 240, and 360 min. Samples for VFA analysis were collected at 6 h, and incubations were continued through 48 h to estimate true digestibility (based on NDF analysis). Increasing the dose of NBPT tended (P < .12) to linearly depress the rate of urea hydrolysis and decreased (P < .0004) subsequent NH3 N formation. Although total VFA concentration at 6 h increased linearly (P < .03), acetate:propionate and estimated true digestibility decreased (P < .01) with increasing NBPT concentration. In Exp. 2, we compared in vitro urea kinetics and digestion of forage-only or mixed forage-grain substrates in response to addition of NBPT. In vitro incubations were conducted in 50-mL test tubes containing either .5 g of ground fescue hay or .5 g of a ground fescue hay and ground corn mixture (50:50, DM basis) to which 0, 6.5, 13, 26, or 52 mg of NBPT in a buffered ruminal fluid innoculum was added. Tubes were incubated in triplicate at 39 degrees C and replicated on consecutive days, with NH3 N and urea concentrations measured at 0, .5, 1, 2, 4, 8, 12, 24, and 48 h. At 48 h, samples for VFA analysis were collected and true digestibility (based on NDF analysis) was estimated. No (P > .10) NBPT dose x substrate interactions were detected. Increasing the dose of NBPT depressed (P < .003) the rate of urea hydrolysis and subsequent NH3 N formation, regardless of substrate. Although total VFA concentration was unaffected (P > .10), the acetate:propionate and estimated true digestibility decreased (P < .002) with higher NBPT addition. In both experiments, the rate of urea degradation was not different (P > .20) from zero for the 26 and 52 mg NBPT treatments, indicating that nearly complete inhibition of urease had been achieved. We conclude that NBPT can be used to reduce the rate of NH3 N release from dietary urea and, thereby, offers the potential to improve nonprotein nitrogen utilization in ruminants. PMID- 10682821 TI - Influence of the novel urease inhibitor N-(n-butyl) thiophosphoric triamide on ruminant nitrogen metabolism: II. Ruminal nitrogen metabolism, diet digestibility, and nitrogen balance in lambs. AB - Three lamb metabolism experiments were conducted to investigate the effects of chronic administration of the novel urease inhibitor N (n-butyl) thiophosphoric triamide (NBPT) on ruminal N metabolism, fermentation, and N balance. In Exp. 1, ruminally cannulated wethers (n = 28; 45.0 +/- .9 kg) were administered one of seven doses of NBPT (0 [control], .125, .25, .5, 1, 2, or 4 g of NBPT daily) and fed a common cracked corn/cottonseed hull-based diet twice daily containing 2% urea at 2.5% of initial BW for the duration of the 15-d experiment. Overall, NBPT decreased (linear P < .0001; quadratic P < .001) ruminal urease activity, resulting in linear increases (P < .0001) in ruminal urea and decreases in ruminal NH3 N concentrations. However, the detection of an NBPT x day interaction (d 2 vs 15; P < .01) indicated that this depression in urea degradation diminished as the experiment progressed. Increasing NBPT linearly decreased (P < .01) total VFA concentrations on d 2 of the experiment, but it had no effect (P > .10) on d 15. Increasing NBPT had no effect (P > .10) on DM or ADF digestibilities, but it linearly decreased (P < .01) N digestibility. Supplementing NBPT produced a linear increase (P < .05) in urinary N excretion and a linear decrease (P < .01) in N retention. In Exp. 2, ruminally cannulated wethers (n = 30; 46.8 +/- .6 kg) were fed one of two basal diets (2.0 vs 1.1% dietary urea) at 2.5% of initial BW and dosed with either 0 (control), .25, or 2 g of NBPT daily for the duration of the 15-d experiment. There were no NBPT x dietary urea interactions (P > .10) for Exp. 2. Increasing NBPT depressed (linear and quadratic P < .0001) ruminal urease activity, producing linear (P < .0001) increases in urea N and linear decreases in NH3 N in the rumen. As in Exp. 1, an NBPT x day interaction (P < .05) was noted for urea, NH3 N, and total VFA concentrations; the maximum response to NBPT occurred on d 2 but diminished by d 15 of the experiment. Administration of NBPT did not influence (P > .10) DM, ADF, or N digestibilities in Exp. 2. In Exp. 3, wether lambs (n = 30; 26.4 +/- .7 kg) were subjected to the same treatment regimen as in Exp. 2 for a 14-d N balance experiment. Although several NBPT x dietary urea interactions (P < .05) were noted, increasing NBPT did not affect (P > .10) N digestibility. Administration of NBPT quadratically increased (P < .10) urinary N excretion, producing a linear decrease (P < .05) in N retention. These results suggest that although NBPT is capable of inhibiting ruminal urease short-term, the ruminal microflora may be capable of adapting to chronic NBPT administration, thereby limiting its practical use in improving the utilization of dietary urea. PMID- 10682822 TI - Metabolize methionine and lysine requirements of growing cattle. AB - Two growth studies were conducted to determine the Met and Lys requirements of growing cattle. In each 84-d trial, steer calves were fed individually diets containing 44% sorghum silage, 44% corn cobs, and 12% supplement (DM basis) at an equal percentage of BW. In Trial 1, 95 crossbred steers (251 kg) were supplemented with urea or meat and bone meal (MBM). Incremental amounts of rumen protected Met were added to MBM to provide 0, .45, .9, 1.35, 3, and 6 g/d metabolizable Met. In Trial 2, 60 steers (210 kg) were supplemented with urea or corn gluten meal (CGM). Incremental amounts of rumen-protected Lys were added to CGM to provide 0, 1, 2, 3, 4, 5, 6, 8, and 10 g/d metabolizable Lys. Supplementation with MBM and CGM increased the supply of metabolizable protein to the animal. Steers fed MBM plus 0 Met gained 49 g/d more than steers fed urea, whereas steers fed CGM plus 0 Lys gained 150 g/d more than steers fed urea. Supplementation of rumen-protected Met and Lys improved ADG in steers fed MBM and CGM, respectively (P < .10). Nonlinear analysis, comparing gain vs supplemental Met and Lys intake, predicted supplemental Met and Lys requirements of 2.9 and .9 g/d, respectively. This amount of additional Met promoted .13 kg/ d gain greater than MBM alone, and this amount of additional Lys promoted .10 kg/d gain greater than the CGM alone. Metabolizable Met and Lys requirements were predicted from Level 1 of NRC (1996) calculated metabolizable protein supply, amino acid analysis of abomasal contents, and the maximum response to supplemental AA. Steers gaining .39 kg/d required 11.6 g/ d Met or 3. 1% of the metabolizable protein requirement, whereas steers gaining .56 kg/d required 22.5 g/d Lys or 5.7% of the metabolizable protein requirement. PMID- 10682823 TI - Evaluation of feather meal as a source of sulfur amino acids for growing steers. AB - In situ and digestion studies were conducted to evaluate feather meal (FTH), blood meal (BM), and meat and bone meal (MBM) for escape protein content, amino acid composition of the escape protein, true protein digestibility, and digestibility of the individual amino acids. Following 12 h of ruminal incubation, escape protein values were 73.5, 92.4, and 60.8% of CP for FTH, BM, and MBM, respectively. Blood meal and MBM were poor sources of sulfur amino acids (SAA), whereas FTH was a good source. Most of the SAA of FTH, however, was Cys, with very little Met. True protein digestibilities were not different for the protein sources (P > .15), ranging from 86.7 to 94.0% of the CP. However, digestibilities of the individual amino acids were quite different. Two growth studies were conducted to evaluate FTH as a source of SAA for growing cattle. The first study used 120 steers (228 +/- 15 kg) supplemented with urea, MBM, MBM plus 1% FTH, or MBM plus 2% FTH. Additionally, incremental amounts of rumen-protected Met were added to treatments containing MBM. Supplementation of MBM increased (P < .05) ADG compared with the urea control. Addition of FTH to MBM resulted in a linear (P < .01) increase in ADG. However, addition of rumen-protected Met to MBM plus FTH treatments further improved gains. Although FTH is an effective source of SAA, Met probably was first-limiting. The second study used 90 steers (243 +/- 18 kg) supplemented with BM plus incremental amounts of SAA from either FTH or rumen-protected Met. Addition of SAA improved ADG compared with BM alone (P < .05). Rumen-protected Met as a source of SAA improved ADG compared with FTH (P < .05). The SAA from FTH promoted a gain response equal to 50% of the response obtained with rumen-protected Met. Formulation of ruminant diets for metabolizable amino acids must account for escape value and digestibility of each individual amino acid. Feather meal is an effective source of SAA; however, Cys supplies over five times the amount supplied by Met. PMID- 10682824 TI - Changes in postprandial plasma and extracellular and ruminal fluid volumes in wethers fed or unfed for 72 hours. AB - Postprandial shifts in body water compartments might limit feed intake by ruminants, especially when an animal becomes partially dehydrated during transportation or other periods of water deprivation. This experiment was conducted to determine the effects of feed and water deprivation on postprandial changes in body water compartments in wethers. Hampshire wethers (n = 8; average BW 42 +/- 2 kg) were used in a crossover design. During each period, four wethers were limit-fed (540 g DM/d: FED) and four were deprived of feed and water for 72 h (DEPRIVED). Wethers were infused i.v. with Evans blue and sodium thiosulfate and intraruminally with Cr- or Co-EDTA, after which blood and ruminal samples were collected for the next 4 h. All wethers were then fed 540 g of feed DM, and infusions were repeated 30 min after feeding. Body water compartment volumes were determined with linear regression using plasma concentrations of Evans blue (plasma volume), and sodium thiosulfate (extracellular volume), and using ruminal fluid concentrations of Cr or Co. Feed and water deprivation decreased (P < .01) extracellular water space but did not affect plasma or ruminal water space. After feeding, extracellular water space decreased (P < .01) and ruminal volume increased (P < .05) in the FED and DEPRIVED wethers. Plasma pools of Na, K, and Mg were not affected by feeding in FED wethers but decreased (P < .05) in DEPRIVED wethers. The increase in ruminal fluid pools of Na, K, and Mg were greater (P < .05) in FED than in DEPRIVED wethers. These results indicate that abnormal water and electrolyte shifts may be factors partially responsible for the decreased feed intake by ruminants subjected to transportation or feed and water deprivation stress. PMID- 10682825 TI - Effects of supplemental degradable intake protein on utilization of medium- to low-quality forages. AB - Three independent experiments were conducted each using 16 ruminally fistulated beef steers fed bermudagrass (8.2% CP, 71% NDF; Exp. 1), bromegrass (5.9% CP, 65% NDF; Exp. 2), or forage sorghum (4.3% CP, 60% NDF; Exp. 3) hays to evaluate the effects of increasing level of supplemental degradable intake protein (DIP) on forage utilization. In each experiment, steers were blocked by weight and assigned to one of four treatments, and hay was offered to each steer at 130% of average voluntary intake for the preceding 5-d period. Supplemental DIP (sodium caseinate) was placed in the rumen at 0700, immediately before feeding forage. Levels of DIP supplementation were .041, .082, and .124% BW; the control received no supplemental DIP. Following a 10-d adaptation, intake and total fecal output were measured for 7 d. In Exp. 1, neither forage OM intake (FOMI) nor fiber (NDF) digestion were influenced (P > or = .20) by increasing level of DIP supplementation. The DIP supplied by the bermudagrass hay was estimated to be 8.2% of the total digestible OM intake (TDOMI) for control steers. In Exp. 2, increasing level of supplemental DIP did not affect (P > or = .26) FOMI but tended to increase total OM intake linearly (TOMI; P = .10). The tendency for a rise in TOMI coupled with a slight numeric increase in digestion resulted in an increase (linear; P = .06) in TDOMI. In the treatment group in which the maximum TDOMI was observed (supplemental DIP treatment of .082% BW), total DIP intake constituted approximately 9.8% of the TDOMI. In Exp. 3, FOMI, TOMI, organic matter digestion (OMD), and TDOMI were improved (P < .01) by increasing amounts of supplemental DIP. Although there was some evidence of a tendency for a decrease in the magnitude of change in TDOMI in response to increasing DIP supplementation, a clear plateau was not achieved with the levels of supplement provided. When the highest level of supplemental DIP was fed, DIP constituted approximately 12.8% of the TDOMI. In conclusion, significant variation was observed among forage in the amount of DIP needed to maximize intake and digestion when expressed in relationship to the digestible OM. PMID- 10682826 TI - Determination of the methionine requirement of growing double-muscled Belgian blue bulls with a three-step method. AB - The three-step technique was used to determine the requirements of total amino acids (TAA) and the first-limiting amino acid (AA) in growing double-muscled Belgian Blue bulls (BBb). In Exp. 1, three double-muscled BBb weighing initially 306 +/- 28 kg received a basal diet consisting of 30% meadow hay and 70% concentrate that was poor in digestible protein but had adequate NE because of continuous infusion of dextrose into the duodenum. The intestinal apparent digestibility of essential AA (EAA) was defined according to their duodenal and ileal flows. It averaged 72% but varied between 60% for Met and 79% for Arg. In Exp. 2, five double-muscled BBb (334 +/- 22 kg) received the same diet supplemented with duodenal infusions of dextrose and four doses of Na-caseinate (28, 56, 84, and 112% of intestinal digestible dietary AA) in a 4 x 4 Latin square design with one additional animal. Nitrogen retention for the basal diet alone and the four increasing supplements of Na-caseinate reached 49, 61, 70, 80, and 86 g/d, respectively. Nitrogen utilization improved from 34.3% without Na caseinate supplementation to a maximum of 40.6%, with the third dose supplying 788 g/d of apparently digestible AA. Based on patterns of plasma concentrations, Met, Phe, and Arg were probably the limiting AA when animals optimized N utilization. In Exp. 3, six double-muscled BBb (315 +/- 25 kg) fed the basal diet received duodenal infusions of dextrose and AA, equivalent to the third dose in Exp. 2, except for digestible Met (9.3, 14.4, 18.4, 22.4, 26.4, and 30.4 g/d) in a 6 x 6 Latin square design. The Met requirement was close to 26.4 g/d on the basis of N retention. PMID- 10682827 TI - Rapid communication: localization of POU1F1 to bovine, ovine, and caprine 1q21 22. PMID- 10682828 TI - Summary of Notifiable Diseases, United States, 1998. AB - The MMWR Summary of Notifiable Diseases, United States, 1998 contains summary tables of the official statistics for the reported occurrence of nationally notifiable diseases in the United States for 1998. These statistics are collected and compiled from reports to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE). PMID- 10682829 TI - Nuclear wavepacket motion producing a reversible charge separation in bacterial reaction centers. AB - The excitation of bacterial reaction centers (RCs) at 870 nm by 30 fs pulses induces the nuclear wavepacket motions on the potential energy surface of the primary electron donor excited state P*, which lead to the fs oscillations in stimulated emission from P* [M.H. Vos, M.R. Jones, C.N. Hunter, J. Breton, J.-C. Lambry and J.-L. Martin (1994) Biochemistry 33, 6750-6757] and in Qy absorption band of the primary electron acceptor, bacteriochlorophyll monomer B(A) [A.M. Streltsov, S.I.E. Vulto, A.Y. Shkuropatov, A.J. Hoff, T.J. Aartsma and V.A. Shuvalov (1998) J. Phys. Chem. B 102, 7293-7298] with a set of fundamental frequencies in the range of 10-300 cm(-1). We have found that in pheophytin modified RCs, the fs oscillations with frequency around 130 cm(-1) observed in the P*-stimulated emission as well as in the B(A) absorption band at 800 nm are accompanied by remarkable and reversible formation of the 1020 nm absorption band which is characteristic of the radical anion band of bacteriochlorophyll monomer B(A)-. These results are discussed in terms of a reversible electron transfer between P* and B(A) induced by a motion of the wavepacket near the intersection of potential energy surfaces of P* and P+B(A)-, when a maximal value of the Franck-Condon factor is created. PMID- 10682830 TI - Induction of tcI 7, a gene encoding a beta-subunit of proteasome, in tobacco plants treated with elicitins, salicylic acid or hydrogen peroxide. AB - We previously isolated, by differential display and 5' RACE (rapid amplification of cDNA ends), cDNAs corresponding to genes activated following cryptogein treatment of tobacco cell suspensions, among them tcI 7 (tcI for tobacco cryptogein Induced), a gene encoding a beta-subunit of proteasome. Here, we report that tcl 7 was up-regulated in tobacco plants treated with elicitins (cryptogein and parasiticein) that have been shown to induce a systemic acquired resistance (SAR). Moreover, subsequent inoculation of tobacco with the pathogen Phytophthora parasitica var. nicotianae (Ppn) was shown to induce an additional activation of tcI 7 in tobacco plants pretreated with cryptogein. We also showed an up-regulation of tcI 7 by salicylic acid (SA). Moreover, accumulation of tcI 7 transcripts after treatment with cryptogein or with SA only occurred in NahG 9 tobacco plants that do not express the salicylate hydroxylase and thus are able to accumulate SA and develop a SAR. Suppressed accumulation of tcI 7 transcripts in NahG 8+ tobacco plants after cryptogein or SA treatment correlated with the loss of SAR. H2O2 was also shown to up-regulate tcI 7 in tobacco plants. Using gene walking by PCR we cloned and sequenced the 5' flanking region of tcI 7 containing hypothetical regulatory sequences, especially myb and NF-kappaB boxes, that could be responsible for the regulation of tcI 7 by salicylic acid and H2O2 respectively. PMID- 10682831 TI - P-glycoprotein is localized in caveolae in resistant cells and in brain capillaries. AB - A significant proportion of P-glycoprotein (P-gp) and caveolin was co-localized in caveolae isolated from resistant (CH(R)C5) cells overexpressing P-gp and from drug-sensitive Chinese hamster ovary cells (AuxB1). The proportion of P-gp and caveolin associated with caveolar microdomains was higher in CH(R)C5 cells grown in the presence of P-gp substrates (cyclosporin A or colchicine) than in untreated CH(R)C5 cells. Coimmunoprecipitation of P-gp and caveolin from CH(R)C5 lysates suggests that there is a physical interaction between them. Furthermore, co-localization of P-gp and caveolin was found in caveolae from brain capillaries, indicating that this association also takes place in vivo. PMID- 10682832 TI - A high affinity nitrate transport system from Chlamydomonas requires two gene products. AB - A nitrate-regulated cluster of genes involved in nitrate transport and assimilation has been identified in Chlamydomonas reinhardtii. Mutant strains of the alga, which are defective in some aspect of transport and assimilation have been used to assign functions to these genes. This analysis has suggested that two gene products are necessary to obtain a functional high affinity nitrate system in Chlamydomonas [Quesada et al. (1994) Plant J. 5, 407-419]. In this paper we have tested this hypothesis by injecting Xenopus oocytes with mRNA prepared from these two cDNAs, Nrt2;1 and Nar2, and then assaying the oocytes for nitrate transport activity. Oocytes injected with single types of mRNA did not show any nitrate transport activity. Furthermore, Nar2 mRNA was toxic to oocytes, with nearly 60%, of the oocytes dead 3 days after the injection. However, when oocytes were injected with a mixture of two mRNAs prepared from Nrt2;1 and Nar2, a high affinity nitrate transport activity could be measured. However, the Km for nitrate of this transport system was 28 microM which is higher than the value of 1.6 microM which had been obtained by the analysis of mutant phenotypes. The pH dependence of the nitrate-elicited currents was consistent with a proton cotransport mechanism. These results prove that two gene products are required to produce a functional high affinity nitrate transport system and that this process does not involve transcriptional regulation. PMID- 10682833 TI - Potential role for triglycerides in signal transduction. AB - We previously reported that endothelin-1 or platelet-derived growth factor promoted in aortic smooth muscle cells a rapid hydrolysis of 1-O-alkyl-2-acyl-sn glycero-3-phosphoethanolamine (alkyl-PE) which was immediately converted into 1-O alkyl-2,3-diacyl-sn-glycerol (alkyl-TG) within 5 s or 60 s respectively [C. Comminges et al. (1996) Biochem. Biophys. Res. Commun. 220, 1008-1013 and C. Comminges et al. (1997) Biochim. Biophys. Acta 1355, 69-80]. In this study, we show that this alkyl-PE hydrolysis is triggered by a transient activation of a specific phospholipase C (PLC) regulated by pertussis toxin-sensitive heterotrimeric G-proteins. Moreover, this PLC can be triggered through a Ca2+ influx depending on L-type Ca2+ channel activation, as suggested by the use of a specific 'activator' S(-)-BayK 8644 and of selective inhibitors such as nimodipine. Interestingly, low concentrations (10(-8)-10(-7)M) of alkyl-TG block the opening of L-type Ca2+ channels, whereas identical concentrations of DG do not alter L-type Ca2+ channels. This study thus unravels a hitherto unrecognized signaling pathway generating alkyl-TG as a novel lipid second messenger, potentially acting as a negative feedback regulator of L-type Ca2+ channels. PMID- 10682834 TI - Functional association between the nef gene product and gag-pol region of HIV-1. AB - Nef gene function is diverse among virus isolates of primate immunodeficiency viruses. We found differential effects of nef mutation on the virus replication between two HIV-1 clones, NL432 and LAI. The nef mutation in NL432 affected the infectivity more severely compared with that in LAI, although the Nef functions of both clones were comparable. Analysis with a series of chimeric viruses between NL432 and LAI revealed that the gag-pol region was responsible for the differential effect of nef mutation. The functional association between Nef and gag-pol suggested that one of the potential targets of Nef was located within the gag-pol region. PMID- 10682835 TI - cDNA-derived amino acid sequence of acetoacetyl-CoA synthetase from rat liver. AB - In order to examine the primary structure of acetoacetyl-CoA synthetase (acetoacetate-CoA ligase, EC 6.2.1.16; AA-CoA synthetase), the cDNA clone encoding this enzyme has been isolated from the cDNA library which was prepared from the liver of rat fed a diet supplemented with 4% cholestyramine and 0.4% pravastatin for 4 days. Nucleotide sequence analysis of cloned cDNA revealed that AA-CoA synthetase of rat liver contains an open reading frame of 2019 nucleotides, and the deduced amino acid sequence (672 amino acid residues) bears 25.0 and 38.9% homologies with acetyl-CoA synthetases of Saccharomyces cerevisiae and Archaeoglobus fulgidus, respectively. PMID- 10682836 TI - Identification of a cDNA encoding an active asparaginyl endopeptidase of Schistosoma mansoni and its expression in Pichia pastoris. AB - Asparaginyl endopeptidases, or legumains, are a recently identified family of cysteine-class endopeptidases. A single gene encoding a Schistosoma mansoni asparaginyl endopeptidase (a.k.a. Sm32 or schistosome legumain) has been reported, but by sequence homology it would be expected to yield an inactive product as the active site C197 had been replaced by N. We now describe a new S. mansoni gene in which C197 is present. Both gene products were expressed in Pichia pastoris. Autocatalytic processing to fully active C197 Sm32 occurred at acid pH. In contrast, N197 Sm32 was not processed and this is consistent with the hypothesis that C197 is essential for catalysis. This was confirmed by mutation of N197 to C and re-expression in Pichia. The availability of recombinant active Sm32 allows detailed analysis of its catalytic mechanism and its function(s) in the biology of this important human parasite. PMID- 10682837 TI - Two Sox9 messenger RNA isoforms: isolation of cDNAs and their expression during gonadal development in the frog Rana rugosa. AB - Sox is a family of SRY-related testis-determining genes. We have isolated two different mRNA isoforms of the frog Sox9 gene from adult frog testis cDNAs. One form (Sox9 alpha) encodes a 482 amino acid protein containing the HMG box, whereas the other form (Sox9 beta), which completely lacks the HMG box, is a truncated 265 amino acid protein of Sox9 alpha. Sox9 alpha is 82% similar to mouse, 86% to chicken, and 77% to trout Sox9 at the amino acid level. Sox9 expression was upregulated in embryos after stage 16, and was seen in both developing testes and ovaries. The size of Sox9 transcripts was determined to be 7.8 knt by Northern blot analysis. In addition, Sox9 alpha expression was found prominently in the testis and brain among various tissues of adult frogs examined, and was considerably higher than Sox9 beta. The fact that Sox9 is expressed in both sexes suggests that this gene is involved in gonadal development of male and female frogs. This is dissimilar to the pattern in birds and mammals, in which Sox9 expression is male-specific. PMID- 10682838 TI - Key amino acids of vasopressin V1a receptor responsible for the species difference in the affinity of OPC-21268. AB - A non-peptide, vasopressin V1a receptor-selective antagonist, OPC-21268, exhibited a markedly higher affinity for the rat V1a receptor (Ki = 380 nM) than for the human V1a receptor (Ki = 140 microM). To delineate the region responsible for the high affinity binding of OPC-21268 for the rat V1a receptor, we have constructed a series of chimeric human and rat V1a receptors, and examined the chimeric and point-mutated receptors by competitive radioligand binding analysis. The results showed that the transmembrane domain (TMD) VI-VII of the vasopressin V1a receptor, in particular the amino acid residue Ala-342 in TMD VII, is the major component conferring the rat-selective binding of OPC-21268 to the V1a receptor. PMID- 10682839 TI - Unusual FTIR and EPR properties of the H2-activating site of the cytoplasmic NAD reducing hydrogenase from Ralstonia eutropha. AB - Soluble NAD-reducing [NiFe]-hydrogenase (SH) from Ralstonia eutropha (formerly Alcaligenes eutrophus) has an infrared spectrum with one strong band at 1956 cm( 1) and four weak bands at 2098, 2088, 2081 and 2071 cm(-1) in the 2150-1850 cm( 1) spectral region. Other [NiFe]-hydrogenases only show one strong and two weak bands in this region, attributable to the NiFe(CN)2(CO) active site. The position of these three bands is highly sensitive to redox changes of the active site. In contrast, reduction of the SH resulted in a shift to lower frequencies of the 2098 cm(-1) band only. These and other properties prompted us to propose the presence of a Ni(CN)Fe(CN)3(CO) active site. PMID- 10682840 TI - Gene structure of human cholecystokinin (CCK) type-A receptor: body fat content is related to CCK type-A receptor gene promoter polymorphism. AB - The transcriptional start site of the human cholecystokinin (CCK)-A receptor gene was determined by the Capsite Hunting method. Two sequence changes were detected, a G to T change in nucleotide -128, and an A to G change in nucleotide -81. The homozygote (T/T, G/G) was detected in 25 of 1296 individuals (1.9%) in the cohort study. This polymorphism showed a significantly higher percent body fat and higher levels of serum insulin and leptin, compared with wild type and heterozygotes. Our study provided the possibility that polymorphism in the promoter region of the CCK-A receptor gene may be one of genetic factors affecting fat deposition. PMID- 10682841 TI - Mutational analysis of phosphorylation sites in the Dictyostelium myosin II tail: disruption of myosin function by a single charge change. AB - The dynamic assembly/disassembly of non-muscle myosin II filaments is critical for the regulation of enzymatic activities and localization. Phosphorylation of three threonines, 1823, 1833 and 2029, in the tail of Dictyostelium discoideum myosin II has been implicated in control of myosin filament assembly. By systematically replacing the three threonines to aspartates, mimicking a phosphorylated residue, we found that position 1823 is the most critical one for the regulation of myosin filament formation and in vivo function. Surprisingly, a single charge change is able to perturb filament formation and in vivo function of myosin II. PMID- 10682842 TI - Requirements for alternative forms of the activator domain, P5abc, in the Tetrahymena ribozyme. AB - The role of P5abc domain of the Tetrahymena LSU self-splicing Group I intron is to enhance the activity of the intron via tertiary interactions involving A-rich bulge and terminal loops L5b and L5c. We constructed and examined alternative forms of the domain that accelerate the ribozymatic reaction. The results indicate that the characteristic structure of P5c subdomain plays an important role by forming L2xL5c interaction (P14) and that the region flanking P5c subdomain can be significantly mutable without much affecting the activity of the ribozyme. PMID- 10682843 TI - Progesterone is a cell death suppressor that downregulates Fas expression in rat corpus luteum. AB - In female rats, apoptotic cell death in the corpus luteum is induced by the prolactin (PRL) surge occurring in the proestrous afternoon during the estrous cycle. We have previously shown that this luteolytic action of PRL is mediated by the Fas/Fas ligand (FasL) system. During pregnancy or pseudopregnancy, apoptosis does not occur in the corpus luteum. Progesterone (P4), a steroid hormone secreted from luteal steroidogenic cells, attenuated PRL-induced apoptosis in cultured luteal cells in a dose-dependent manner. P4 significantly decreased the expression of mRNA of Fas, but not FasL, in cultured luteal cells prepared from both proestrous and mid-pseudopregnant rats. These data indicate that P4 suppresses PRL-induced luteal cell apoptosis via reduction of the expression level of Fas mRNA in the corpus luteum, suggesting that P4 acts as an important factor that can change the sensitivity of corpus luteum to PRL. PMID- 10682844 TI - Closed loops of nearly standard size: common basic element of protein structure. AB - By screening the crystal protein structure database for close Calpha-Calpha contacts, a size distribution of the closed loops is generated. The distribution reveals a maximum at 27+/-5 residues, the same for eukaryotic and prokaryotic proteins. This is apparently a consequence of polymer statistic properties of protein chain trajectory. That is, closure into the loops depends on the flexibility (persistence length) of the chain. The observed preferential loop size is consistent with the theoretical optimal loop closure size. The mapping of the detected unit-size loops on the sequences of major typical folds reveals an almost regular compact consecutive arrangement of the loops. Thus, a novel basic element of protein architecture is discovered; structurally diverse closed loops of the particular size. PMID- 10682845 TI - p97 ATPase, an ATPase involved in membrane fusion, interacts with DNA unwinding factor (DUF) that functions in DNA replication. AB - DNA unwinding factor (DUF) unwinds duplex DNA and is supposed to function in DNA replication in Xenopus egg extracts. Here we report the isolation and analysis of a DUF-interacting factor. By immunoprecipitation, we found that p97 ATPase (p97) interacts with DUF in Xenopus egg extracts. This interaction was confirmed by the in vitro binding of purified p97 with DUF. When sperm chromatin was added to Xenopus egg extracts to construct nuclei active in DNA replication, p97 was incorporated into the nuclei. These data suggest that the complex of DUF and p97 may function in DNA replication. PMID- 10682846 TI - Chitosan-induced phospholipase A2 activation and arachidonic acid mobilization in P388D1 macrophages. AB - We have found that chitosan, a polysaccharide present in fungal cell walls, is able to activate macrophages for enhanced mobilization of arachidonic acid in a dose- and time-dependent manner. Studies aimed at identifying the intracellular effector(s) implicated in chitosan-induced arachidonate release revealed the involvement of the cytosolic Group IV phospholipase A2 (PLA2), as judged by the inhibitory effect of methyl arachidonoyl fluorophosphonate but not of bromoenol lactone. Interestingly, priming of the macrophages with lipopolysaccharide renders the cells more sensitive to a subsequent stimulation with chitosan, and this enhancement is totally blocked by the secretory PLA2 inhibitor 3-(3 acetamide)-1-benzyl-2-ethylindolyl-5-oxy-propanesulfonic acid (LY311727). Collectively, the results of this work establish chitosan as a novel macrophage activating factor that elicits AA mobilization in P388D1 macrophages by a mechanism involving the participation of two distinct phospholipases A2. PMID- 10682847 TI - Activation of recombinant proenteropeptidase by duodenase. AB - Duodenase, a serine proteinase from bovine Brunner's (duodenal) glands that was predicted to be a natural activator of enteropeptidase zymogen, cleaves and activates recombinant single-chain bovine proenteropeptidase (kcat/Km = 2700 M( 1) s(-1)). The measured rate of proenteropeptidase cleavage by duodenase was about 70-fold lower compared with the rate of trypsin-mediated cleavage of the zymogen. The role of duodenase is supposed to be the primary activator of proenteropeptidase maintaining a certain level of active enteropeptidase in the duodenum. A new scheme of proteolytic activation cascade of digestive proteases is discussed. PMID- 10682848 TI - Role of the highly conserved Asp-Arg-Tyr motif in signal transduction of the CB2 cannabinoid receptor. AB - The DRY motif, at the junction of transmembrane helix 3 and intracellular loop 2 of G protein-coupled receptors, is highly conserved. Mutations were introduced into the CB2 cannabinoid receptor to study the role of this motif in CB2 signaling. D mutations (DRY130-132AAA and D130A) markedly reduced binding of cannabinoid agonists, while no significant reduction was observed with R131A or Y132A. Mutating R (R131A) only partially reduced, and mutating Y (Y132A) more efficiently reduced the cannabinoid-induced inhibition of adenylyl cyclase. Thus, in CB2, D130 is involved in agonist binding, whereas Y seems to have a role in receptor downstream signaling. PMID- 10682849 TI - Virus-induced permeability transition in mitochondria. AB - Isolated rat liver mitochondria undergo permeability transition after supplementation with a suspension of tobacco mosaic virus. Four mitochondrial parameters proved the opening of the permeability transition pore in the inner mitochondrial membrane: increased oxygen consumption, collapse of the membrane potential, release of calcium ions from mitochondria, and high amplitude mitochondrial swelling. All virus-induced changes in mitochondria were prevented by cyclosporin A. These effects were not observed if the virus was treated with EGTA or disrupted by heating. Protein component of the virus particle in the form of 20S aggregate A-protein, or helical polymer, as well as supernatant of the heat-disrupted virus sample, had no effect on mitochondrial functioning. Electron microscopy revealed the direct interaction of the virus particles with isolated mitochondria. The possible role of the mitochondrial permeability transition pore in virus-induced apoptosis is discussed. PMID- 10682850 TI - Characterization of the c-specific promoter of the gene encoding human endothelin converting enzyme-1 (ECE-1). AB - Human ECE-1 is expressed in four isoforms with different tissue distribution and its mRNA and protein levels are altered under certain pathophysiological conditions. To investigate the transcriptional regulation of ECE-1, we studied the regulatory region of ECE-1c, the major ECE-1 isoform. A genomic clone comprising the complete human ECE-1 gene including the putative ECE-1c-specific promoter was obtained. Up to 968 bp upstream of the putative c-specific translation initiation start codon and several serial deletion mutants were subcloned into a reporter vector and transfected into endothelial (BAEC, EA.hy926, ECV304) and epithelial (MDA MB435S, MCF7) cells, showing very strong promoter activity in comparison to the SV40 promoter and to the previously described ECE-1a and 1b promoters. Transfection of serial deletion mutants indicated two positive regulatory regions within the promoter (-142/-240 and 240/490) likely involved in binding GATA and ETS transcription factors. RNase protection assay (RPA) and 5'-RACE revealed multiple transcriptional start sites located at about -110, -140 and -350 bp. Site-directed mutagenesis demonstrated a crucial role for the E2F cis-element for basal ECE-1c promoter activity. Additionally, we found a correlation between isoform-specific ECE-1 mRNA levels and corresponding ECE-1a, 1b, 1c promoter activities. PMID- 10682851 TI - Mutagenic studies on human protein disulfide isomerase by complementation of Escherichia coli dsbA and dsbC mutants. AB - Protein disulfide isomerase (PDI) exhibits both an oxido-reductase and an isomerase activity on proteins containing cysteine residues. These activities arise from two active sites, both of which contain pairs of redox active cysteines. We have developed two simple in vivo assays for these activities of PDI, based on the demonstration that PDI can complement both a dsbA mutation and a dsbC mutation when expressed to the periplasm of Escherichia coli. We constructed a variety of mutants in and around the active sites of PDI and analysed them using these complementation assays. Our analysis showed that the active site amino acid residues have a major role in determining the activities exhibited by PDI, particularly the N-terminal cysteine of the N-terminal active site. The roles of the histidine residue at position 38 and the glutamic acid residue at position 30 were also studied using these assays. The results show that these two in vivo assays should be useful for rapid screening of mutants in PDI prior to purification and detailed biochemical analysis. PMID- 10682852 TI - The effect of reactive oxygen species generated from the mitochondrial electron transport chain on the cytochrome c oxidase activity and on the cardiolipin content in bovine heart submitochondrial particles. AB - The effect of reactive oxygen species (ROS), produced by the mitochondrial respiratory chain, on the activity of cytochrome c oxidase and on the cardiolipin content in bovine heart submitochondrial particles (SMP) was studied. ROS were produced by treatment of succinate-respiring SMP with antimycin A. This treatment resulted in a large production of superoxide anion, measured by epinephrine method, which was blocked by superoxide dismutase (SOD). Exposure of SMP to mitochondrial mediated ROS generation, led to a marked loss of cytochrome c oxidase activity and to a parallel loss of cardiolipin content. Both these effects were completely abolished by SOD+catalase. Added cardiolipin was able to almost completely restore the ROS-induced loss of cytochrome c oxidase activity. No restoration was obtained with peroxidized cardiolipin. These results demonstrate that mitochondrial mediated ROS generation affects the activity of cytochrome c oxidase via peroxidation of cardiolipin which is needed for the optimal functioning of this enzyme complex. These results may prove useful in probing molecular mechanism of ROS-induced peroxidative damage to mitochondria which have been proposed to contribute to aging, ischemia/reperfusion and chronic degenerative diseases. PMID- 10682853 TI - Expression of GIRK (Kir3.1/Kir3.4) channels in mouse fibroblast cells with and without beta1 integrins. AB - G protein-activated K+ channel (GIRK) subunits possess a conserved extracellular integrin-binding motif (RGD) and bind directly to beta1 integrins. We expressed GIRK1/GIRK4 channels labeled with green fluorescent protein in fibroblast cell lines expressing or lacking beta1 integrins. Neither plasma membrane localization nor agonist-evoked GIRK currents were affected by the absence of beta1 integrins or by incubation with externally applied RGD-containing peptide. Mutation of the aspartate (D) of RGD impaired currents, GIRK glycosylation, and membrane localization, but the interaction with beta1 integrins remained intact. Thus, beta1 integrins are not essential for functional GIRK expression; and the GIRK integrin interactions involve structural elements other than the RGD motif. PMID- 10682854 TI - NMR structure of the channel-former zervamicin IIB in isotropic solvents. AB - Spatial structure of the membrane channel-forming hexadecapeptide, zervamicin IIB, was studied by NMR spectroscopy in mixed solvents of different polarity ranging from CDCl3/CD3OH (9:1, v/v) to CD3OH/H2O (1:1, v/v). The results show that in all solvents used the peptide has a very similar structure that is a bent amphiphilic helix with a mean backbone root mean square deviation (rmsd) value of ca. 0.3 A. Side chains of Trp1, Ile2, Gln3, Ile5 and Thr6 are mobile. The results are discussed in relation to the validity of the obtained structure to serve as a building block of zervamicin IIB ion channels. PMID- 10682855 TI - Isolation and expression of a novel alternatively spliced mu opioid receptor isoform, MOR-1F. AB - The MOR-1 gene is large, with a recent study reporting nine exons spanning 250 kb which combine to yield six different mu opioid receptor splice variants. We now report the isolation of exon 10, which is contained within yet another splice variant, MOR-1F, which is composed of exons 1, 2, 3, 10, 6, 7, 8 and 9. Exon 10 comprises 186 bp which predict a unique 58 amino acid sequence extending beyond exon 3. It has been mapped between exons 4 and 6 and has flanking consensus splice sequences. On Northern blot analysis, the MOR-1F mRNA is smaller than the other MOR-1 mRNAs. When expressed in CHO cells, MOR-1F binds the mu opioid radioligand [3H]DAMGO with high affinity (K(D) = 1.04+/-0.03 nM). Competition studies demonstrated the selectivity of the variant for mu opioid ligands, supporting its classification within the mu opioid receptor family. PMID- 10682856 TI - Spin label EPR structural studies of the N-terminus of alpha-spectrin. AB - Spectrin, a vital component in human erythrocyte, is composed of alpha- and beta subunits, which associate to form (alphabeta)2 tetramers. The tetramerization site is believed to involve the alpha-spectrin N-terminus and the beta-spectrin C terminus. Abnormal interactions in this region may lead to blood disorders. It has been proposed that both termini consist of partial structural domains and that tetramerization involves the association of these partial domains. We have studied the N-terminal region of a model peptide for alpha-spectrin by making a series of double spin-labeled peptides and studying their dipolar interaction by electron paramagnetic resonance methods. Our results indicate that residues 21-42 of the N-terminus region exhibit an alpha-helical conformation, even in the absence of B-spectrin. PMID- 10682857 TI - Myosin light chain phosphorylation-dependent modulation of volume-regulated anion channels in macrovascular endothelium. AB - The Rho/Rho-associated kinase (ROK) pathway has been shown to modulate volume regulated anion channels (VRAC) in cultured calf pulmonary artery endothelial (CPAE) cells. Since Rho/ROK can increase myosin light chain phosphorylation, we have now studied the effects of inhibitors of myosin light chain kinase (MLCK) or myosin light chain phosphatase (MLCP) on VRAC in CPAE. Application of ML-9, an MLCK inhibitor, inhibited VRAC, both when applied extracellularly or when dialyzed into the cell. A similar inhibitory effect was obtained by dialyzing the cells with AV25, a specific MLCK inhibitory peptide. Conversely, NIPP1(191-210), an MLCP inhibitory peptide, potentiated the activation of VRAC by a 25% hypotonic stimulus. These data indicate that activation of VRAC is modulated by MLC phosphorylation. PMID- 10682858 TI - pH control of the plant outwardly-rectifying potassium channel SKOR. AB - SKOR, an Arabidopsis depolarisation-activated K+-selective channel, was expressed in Xenopus oocytes, and external and internal pH effects were analysed. Internal pH was manipulated by injections of alkaline or acidic solutions or by acid load from acetate-containing medium. An internal pH decrease from 7.4 to 7.2 induced a strong (ca. 80%) voltage-independent decrease of the macroscopic SKOR current, the macroscopic gating parameters and the single channel conductance remained unchanged. An external acidification from 7.4 to 6.4 had similar effects. It is proposed that pH changes regulate the number of channels available for activation. Sensitivity of SKOR activity to pH in the physiological range suggests that internal and external pH play a role in the regulation of K+ secretion into the xylem sap. PMID- 10682859 TI - The platelet cytoskeleton regulates the aggregation-dependent synthesis of phosphatidylinositol 3,4-bisphosphate induced by thrombin. AB - Pretreatment of intact platelets with cytochalasin D prevented actin polymerization and cytoskeleton reorganization induced by thrombin, but did not affect platelet aggregation. Under these conditions, synthesis of phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P2) stimulated by thrombin was strongly inhibited, while production of phosphatidic acid was unaffected. The inhibitory effect of cytochalasin D was not observed when platelet aggregation was prevented by the RGDS peptide. We also found that cytochalasin D did not affect PtdIns(3,4)P2 synthesis induced by concanavalin A (ConA), which is known to occur through an aggregation-independent mechanism. Moreover, thrombin, but not ConA, induced the translocation of phosphatidylinositol 3-kinase to the cytoskeleton. This process was equally inhibited by both the RGDS peptide and cytochalasin D. These results demonstrate that the cytoskeleton represents a functional link between thrombin-induced aggregation and synthesis of PtdIns(3,4)P2. PMID- 10682860 TI - Eukaryotic selenocysteine tRNA has the 9/4 secondary structure. AB - There are two secondary structure models for the eukaryotic selenocysteine (Sec) tRNA(Sec). One model, the 9/4 structure, was experimentally tested and possesses acceptor and T-stems with 9 and 4 bp, respectively [Sturchler et al., 1993; Hubert et al., 1998]. The other one, the 7/5 secondary structure with a bulge in the T-stem, was derived from theoretical calculation [Ioudovitch and Steinberg, 19991. In this report, we show more experimental results supporting the 9/4 secondary structure. Several tRNA(Sec) mutants, whose secondary structure can adopt only the 9/4 structure, were active for serylation and selenylation. Some mutants that cannot base-pair between positions 26 and 44 to provide the 6 bp anticodon stem were still active, inconsistent with the model by Steinberg. We also show that the orientation of the V-arm directly or indirectly influences the selenylation activity, and that the rigid 6 bp D-stem is important. Finally, we conclude that all tRNA(Sec) possess the 13 bp domain II made by the stacking of the colinear AA and T-stems, whether they present the 9/4 structure in Eukarya and Archaea or the 8/5 structure in bacteria. PMID- 10682861 TI - Involvement of asparagine 118 in the nucleotide specificity of the catalytic subunit of protein kinase CK2. AB - Protein kinase CK2 is a heteromeric enzyme with catalytic (alpha) and regulatory (beta) subunits which form an alpha2beta2 holoenzyme and utilizes both ATP and GTP as nucleotide substrate. Site-directed mutagenesis of CK2alpha subunit was used to study this capacity to use GTP. Deletion of asparagine 118 (alpha(deltaN118)) or the mutant alphaN118E gives a 5-6-fold increase in apparent Km for GTP with little effect on the affinity for ATP. Mutants alphaN118A and alphaD120N did not alter significantly the Km for either nucleotide. CK2alphaN118 has an apparent Ki for inosine 5' triphosphate 5-fold higher than wild-type and is very heat labile. These studies complement recent crystallographic data indicating a role for CK2alpha asparagine 118 in binding the guanine base. PMID- 10682862 TI - Identification of a melanoma antigen, PRAME, as a BCR/ABL-inducible gene. AB - In order to elucidate molecular events in BCR/ABL-induced transformation, we adopted a polymerase chain reaction (PCR)-based technique of differential display and compared mRNA expression in human factor-dependent cells, TF-1, with that in factor-independent cells, ID-1, which were established from TF-1 cells by transfection of BCR/ABL. Cloning and sequencing of a gene which was upregulated in ID-1 cells revealed that the gene was identical to a melanoma antigen, PRAME. Our present study demonstrated that PRAME was markedly expressed in primary leukemic cells with chronic myeloid leukemia (CML) in blastic crisis and Philadelphia (Ph)+-acute lymphoblastic leukemia (ALL), in which BCR/ABL played an important role as a pathogenic gene. Moreover, comparison of PRAME expression among CD34+ cells with CML in blastic, accelerated, and chronic phases revealed a higher expression in CML in advanced phases. Thus PRAME was considered to be a good candidate for a marker of Ph+-leukemic blast cells as well as a new target antigen of leukemic blast cells that cytotoxic T cells can recognize. PMID- 10682863 TI - Mutagenesis of the proposed iron-sulfur cluster binding ligands in Escherichia coli biotin synthase. AB - Biotin synthase (BioB) is a member of a family of enzymes that includes anaerobic ribonucleotide reductase and pyruvate formate lyase activating enzyme. These enzymes all use S-adenosylmethionine during turnover and contain three highly conserved cysteine residues that may act as ligands to an iron-sulfur cluster required for activity. Three mutant enzymes of BioB have been made, each with one cysteine residue (C53, 57, 60) mutated to alanine. All three mutant enzymes were inactive, but they still exhibited the characteristic UV-visible spectrum of a [2Fe-2S]2+ cluster similar to that of the wild-type enzyme. PMID- 10682864 TI - Regulation of the cardiac voltage-gated Na+ channel (H1) by the ubiquitin-protein ligase Nedd4. AB - The cardiac voltage-gated Na+ channel H1, involved in the generation of cardiac action potential, contains a C-terminal PY motif (xPPxY). Since PY motifs are known ligands to WW domains, we investigated their role for H1 regulation and the possible involvement of the WW domain containing ubiquitin-protein ligase Nedd4, taking advantage of the Xenopus oocyte system. Mutation of the PY motif leads to higher peak currents when compared to wild-type channel. Moreover, co-expression of Nedd4 reduced the peak currents, whereas an enzymatically inactive Nedd4 mutant increased them, likely by competing with endogenous Nedd4. The effect of Nedd4 was not observed in the PY motif mutated channel or in the skeletal muscle voltage-gated Na+ channel, which lacks a PY motif. We conclude that H1 may be regulated by Nedd4 depending on WW-PY interaction, and on an active ubiquitination site. PMID- 10682865 TI - Suppression of GD1alpha ganglioside-mediated tumor metastasis by liposomalized WHW-peptide. AB - GD1alpha ganglioside-replica peptides were recently isolated from a phage displayed random pentadecapeptide library by assaying for inhibition of adhesion of RAW117-H10 lymphosarcoma cells to hepatic sinusoidal microvessel endothelial (HSE) cells. We show here that the Trp-His-Trp (WHW) peptide was identified as a minimal sequence of the GD1alpha-replica peptide WHWRHRIPLQLAAGR. The addition of WHW peptide-attached liposomes displayed efficient inhibition of liver metastasis of RAW117-H10 cells as well as of GD1alpha-mediated adhesion of RAW117-H10 cells to HSE cells in vitro. These results suggest that engineered liposomes for peptide delivery are applicable to treatment for metastasis. PMID- 10682866 TI - The N-terminal domain of the light-harvesting chlorophyll a/b-binding protein complex (LHCII) is essential for its acclimative proteolysis. AB - Variations in the amount of the light-harvesting chlorophyll a/b-binding protein complex (LHCII) is essential for regulation of the uptake of light into photosystem II. An endogenous proteolytic system was found to be involved in the degradation of LHCII in response to elevated light intensities and the proteolysis was shown to be under tight regulation [Yang, D.-H. et al. (1998) Plant Physiol. 118, 827-834]. In this study, the substrate specificity and recognition site towards the protease were examined using reconstituted wild-type and mutant recombinant LHCII. The results show that the LHCII apoprotein and the monomeric form of the holoprotein are targeted for proteolysis while the trimeric form is not. The N-terminal domain of LHCII was found to be essential for recognition by the regulatory protease and the involvement of the N-end rule pathway is discussed. PMID- 10682867 TI - Subcellular localization and processing of the lytic transglycosylase of the conjugative plasmid R1. AB - Protein P19 encoded by the conjugative resistance plasmid R1, is essential for efficient conjugative DNA transfer and infection by the pilus-specific RNA phage R17. Based on sequence homologies P19 belongs to a family of lysozyme-like virulence factors which are found in type III and type IV secretion systems. In this report we describe the processing and subcellular localization of P19. Pulse chase experiments were used to demonstrate the processing of P19 by the signal peptidase I of Escherichia coli. Translocation of P19 across the inner membrane was shown by gene 19-phoA fusions. Cell fractionation studies of P19 expressing cells showed the presence of P19 in the membrane compartment. P19 was solubilized with the detergent Sarkosyl indicating an inner membrane localization. Using sucrose density gradient centrifugation to separate inner and outer membranes, P19 was found in both membrane fractions. Taken together, our data suggest that mature P19 is a periplasmic protein which may be attached to the proposed membrane-spanning DNA transport complex. PMID- 10682868 TI - Nutrition on the net. PMID- 10682869 TI - The potential for dietary supplements to reduce premenstrual syndrome (PMS) symptoms. AB - Many types of dietary supplements have been advocated for the reduction of certain symptoms of premenstrual syndrome (PMS). However, only one supplement calcium-has been demonstrated to be of significant benefit in a large, rigorous, double-blind, placebo-controlled trial. Limited evidence suggests that magnesium, vitamin E and carbohydrate supplements might also be useful, but additional research is needed to confirm these findings. Trials of vitamin B6 supplementation have had conflicting results, and high doses of this vitamin taken for prolonged periods of time can cause neurological symptoms. Trials of evening primrose oil have also had conflicting results; the two most rigorous studies showed no evidence of benefit. A variety of herbal products are suggested to reduce symptoms of PMS. The efficacy of these products is uncertain because of a lack of consistent data from scientific studies. Health professionals should be aware of the possible use of these supplements and ask those with PMS about their use of such products and counsel them based upon the totality of evidence. PMID- 10682870 TI - Preliminary observation: oral zinc sulfate replacement is effective in treating muscle cramps in cirrhotic patients. AB - BACKGROUND: While not life threatening, muscle cramps severely affect the quality of life of patients with cirrhosis. AIM: To determine whether oral zinc sulfate therapy decreases the frequency and severity of muscle cramps in cirrhotic patients. METHODS: 12 patients with cirrhosis (5 Child's A, 3 Child's B, and 4 Child's C), hypozincemia and muscle cramps at least thrice weekly received oral zinc sulfate 220 mg BID for 12 weeks. Patients answered a questionnaire regarding their muscle cramps symptoms at the beginning and end of the study. RESULTS: Muscle cramps occurred in all patients at rest, mainly while sleeping (8/12), and in two patients also during exercise. Cramps were located in calves (10/12), feet (4/12) and hands (4/12) more commonly. Zinc supplementation improved cramps in 10/12 patients, and in seven of these patients the cramps completely resolved. One patient experienced mild watery diarrhea that resolved upon discontinuation of the zinc sulfate. No other complication of zinc supplementation was noted. CONCLUSION: A potential relationship between zinc deficiency and muscle cramps in the setting of cirrhosis has not been suggested before. Zinc supplementation may lead to improvement in symptoms associated with muscle cramps in cirrhosis. PMID- 10682871 TI - Influence of purine intake on uric acid excretion in infants fed soy infant formulas. AB - OBJECTIVE: These studies tested the hypothesis that increasing intake of purines, delivered as RNA from soy protein-based infant formula, would increase urinary uric acid excretion in infants. METHODS: Study One examined the influence of feeding on serum uric acid in a total of 178 infants from four separate trials with infants fed commercial and experimental soy-based and milk-based infant formulas or human milk. Studies Two and Three compared the effect of a standard purine soy formula (STD Purine; 180 mg purines/L from RNA) and a reduced purine soy formula (Reduced Purine; 65 mg purines/L; 26 mg/L from RNA and 39 mg/L from ribonucleotides) on urinary uric acid excretion in infants. In Study Two, 11 infants ranging in age from 16 to 128 days of age were fed both formulas in a random crossover design. Complete 72-hour urine collections were done at the end of each 11-day feeding period. Urinary uric acid excretion was expressed as mmol/day. In Study Three, 33 infants were enrolled before eight days of age and randomized to one of the formulas one week later. Spot urine samples were collected at 28 and/or 56 days of age and urinary uric acid concentration was expressed as mmol/mmol creatinine. RESULTS: In Study One, each of the feedings resulted in mean serum uric acid levels within normal reference ranges. Soy formula led to higher serum uric acid levels than human milk, and human milk to levels indistinguishable from cow milk-based formulas. In Study Two, infants excreted significantly more uric acid in the urine when fed the STD Purine formula compared to the Reduced Purine formula (0.86+/-.04 vs. 0.57+/-.04 mmol/d) (p = 0.006). In Study Three, infants fed the STD Purine formula had a significantly higher concentration of uric acid in their urine compared to those fed the Reduced Purine formula (2.1+/-0.2 vs. 1.4+/-0.1 mmol uric acid/mmol creatinine) (p = 0.0001). CONCLUSION: These data indicate that healthy infants can digest RNA and subsequently absorb the liberated purine ribonucleotides as determined by urinary uric acid concentration. PMID- 10682872 TI - Low-density lipoprotein subclass distribution pattern and adiposity-associated dyslipidemia in postmenopausal women. AB - OBJECTIVE: A predominance of small, dense low-density lipoprotein (LDL) particles (subclass pattern B) is associated with increased risk for coronary heart disease and is characterized by elevated triglycerides and depressed high-density lipoprotein (HDL) cholesterol concentrations. The present analysis was undertaken to assess the impact of LDL subclass distribution pattern and adiposity on serum lipids in postmenopausal women. METHODS: Anthropometric measurements and fasting lipid data were obtained from 254 postmenopausal women 70 years of age or younger, not receiving sex hormone replacement, who were participating in a clinical trial designed to assess the influence of hormone replacement regimens on coronary heart disease risk markers. RESULTS: The prevalence of LDL subclass pattern B was 32%. Triglyceride levels were higher and HDL cholesterol lower (both p<0.001) in women with pattern B vs. pattern A, but total and LDL cholesterol levels did not differ. LDL subclass pattern contributed independently to the variance in HDL cholesterol (p<0.001) and log(e) triglyceride (p<0.001) concentrations explained by anthropometric variables (waist circumference or body mass index). Compared to women with LDL subclass pattern A and waist circumference below the median value of 83.0 centimeters, those with pattern B and waist > or =83.0 centimeters had markedly lower HDL cholesterol levels [44.0 (41.6-47.4) vs. 57.2 (54.1-60.3) mg/dL, mean (95% CI)] and increased triglyceride concentrations [geometric mean 147.8 (131.6-165.7) vs. 95.4 (88.2-102.5) mg/dL]. CONCLUSIONS: These data suggest that adiposity and LDL subclass distribution pattern are independent determinants of plasma triglyceride and HDL cholesterol concentrations in postmenopausal women. PMID- 10682873 TI - The interaction between dietary fructose and magnesium adversely affects macromineral homeostasis in men. AB - OBJECTIVE: Studies with rats have found that an interaction between fructose and magnesium affects macromineral metabolism; high dietary fructose significantly increased kidney calcification in both male and female rats, particularly when dietary magnesium was low. This study tests the hypothesis that an interaction between dietary fructose and magnesium adversely affects macromineral homeostasis in men. METHODS: Eleven men aged 22 to 40 years were fed a mixed, Western diet for four 42-day dietary periods in which dietary magnesium was either approximately 170 or 370 mg/day and dietary fructose was either 4% or 20% of energy. A decaffeinated beverage containing high fructose corn syrup replaced cornstarch, bread and rice in the low fructose diet to give the high fructose diet. RESULTS: High dietary fructose significantly (p<0.01) increased magnesium balance during both low and high dietary magnesium intakes. Ultrafilterable and ionized serum magnesium also apparently were related to magnesium and fructose intakes; they were higher when fructose was fed and when Mg intakes were high. High fructose depressed calcium balance: the effect tended to be more marked when dietary Mg was low. High dietary fructose also significantly (p<0.005) decreased phosphorous balance. Urinary phosphorous losses were significantly (p<0.001) higher when high dietary fructose was fed. High dietary fructose also increased the concentration of serum alkaline phosphatase (p<0.005). CONCLUSION: These findings indicate that dietary fructose adversely affects macromineral homeostasis in humans and suggest further studies to see if a high fructose diet coupled with low dietary magnesium and marginal calcium leads to bone loss. PMID- 10682874 TI - A noninvasive measure of physical maturity as a predictor of bone mass in children. AB - OBJECTIVE: The purpose of this study was to describe the accumulation of whole body bone mass in a longitudinal study of prepubertal boys and girls using Roche's physical maturity index as a measure of developmental age. METHODS: We measured 561 children (39% white, 61% African-American) from a suburban school district, representing an ethnically mixed, middle-class community adjacent to Detroit. Anthropometric measures taken for the present study included recumbent length (cm), stature (cm), weight (kg), whole body bone mineral content (WBBMC in g) and a noninvasive measure of physical maturity (PM%). PM% was calculated from published formulae derived from data from the Fels Longitudinal Study, using recumbent length, weight, midparental stature, age, and age- and gender-specific regression coefficients. RESULTS: At average age 9.9 (+/-0.6) years, there were no significant gender differences in stature, recumbent length, weight, or WBBMC in either ethnic group. Average PM for girls was significantly greater than that for boys within each ethnic group. There were no significant ethnic differences in PM in either gender. Stature and WBBMC were significantly different in the two ethnic groups for both boys and girls. Regressions of WBBMC on PM and chronological age indicated that PM explained more of the variance in WBBMC than did age (r2 ranging from 0.28 to 0.75 for PM versus 0.01 to 0.06 for age). In the case of African-American boys, r2 was similar (0.09 for PM and 0.06 for age). CONCLUSIONS: PM is a useful, noninvasive measure of developmental age that is significantly correlated with bone mass in children. Our study also indicates that PM is a better predictor of WBBMC than chronological age. Because PM can be calculated without using invasive and potentially expensive methods, PM may be useful in some clinical as well as research settings. PMID- 10682875 TI - The effects of varying dietary fat on the nutrient intake in male and female runners. AB - OBJECTIVE: The present study examined the effects of varying dietary fat levels on nutrients in female and male endurance runners. METHODS: Three diets (low, medium and high fat) were designed for each subject using their food preferences and three-day food records. Each diet was eaten for 28 to 31 days. The diets were self-selected from seven-day sample menus. Twelve male and 13 female runners between 18 and 55 years of age who averaged 42 miles/week participated in the study. Daily food intakes, activity records and weekly palatability/hunger scales were completed. RESULTS: Dietary fat intakes, as a percent of total energy intake (%E), averaged 17%E, 31%E, and 44%E on the low, medium and high fat diets, respectively. Energy consumption was less than their estimated energy expenditure (EEE) on all diets. On the low fat diet, the female runners were consuming approximately 60% of their EEE. As dietary fat increased, the difference between calorie intake and estimated energy expenditure became less and the subjects were less hungry on the two higher fat diets. For all subjects, as energy intakes increased, so did carbohydrate intake. Therefore, carbohydrate intake was not different on the two lower fat diets. Irrespective of gender, calcium and zinc intakes, which were below 1989 RDAs, increased with increasing fat intakes, between the low and medium fat diets. Zinc intake was also higher on the highest fat diet. Essential fatty acid intakes for females on the low fat diet were less than 2.5%E. Half of the female runners ate less than the RDA of calcium and zinc on the low fat diet and Fe on the medium fat diet. CONCLUSION: This study suggests that endurance runners may not be consuming enough calories on a low fat diet and that increasing dietary fat increased energy consumption. On the low fat diet, essential fatty acids and some minerals (especially zinc) may be too low. A low fat diet could compromise health and performance. PMID- 10682876 TI - The effects of varying dietary fat on performance and metabolism in trained male and female runners. AB - OBJECTIVES: Low dietary fat intake has become the diet of choice for many athletes. Recent studies in animals and humans suggest that a high fat diet may increase VO2max and endurance. We studied the effects of a low, medium and high fat diet on performance and metabolism in runners. METHODS: Twelve male and 13 female runners (42 miles/week) ate diets of 16% and 31% fat for four weeks. Six males and six females increased their fat intakes to 44%. All diets were designed to be isocaloric. Endurance and VO2max were tested at the end of each diet. Plasma levels of lactate, pyruvate, glucose, glycerol, and triglycerides were measured before and after the VO2max and endurance runs. Free fatty acids were measured during the VO2max and endurance runs. RESULTS: Runners on the low fat diet ate 19% fewer calories than on the medium or high fat diets. Body weight, percent body fat (males=71 kg and 16%; females=57 kg and 19%), VO2max and anaerobic power were not affected by the level of dietary fat. Endurance time increased from the low fat to medium fat diet by 14%. No differences were seen in plasma lactate, glucose, glycerol, triglycerides and fatty acids when comparing the low versus the medium fat diet. Subjects who increased dietary fat to 44% had higher plasma pyruvate (46%) and lower lactate levels (39%) after the endurance run. CONCLUSION: These results suggest that runners on a low fat diet consume fewer calories and have reduced endurance performance than on a medium or high fat diet. A high fat diet, providing sufficient total calories, does not compromise anaerobic power. PMID- 10682877 TI - A diet high in whole and unrefined foods favorably alters lipids, antioxidant defenses, and colon function. AB - OBJECTIVE: Diets rich in whole and unrefined foods, like whole grains, dark green and yellow/orange-fleshed vegetables and fruits, legumes, nuts and seeds, contain high concentrations of antioxidant phenolics, fibers and numerous other phytochemicals that may be protective against chronic diseases. This study compared the effects of a phytochemical-rich diet versus a refined-food diet on lipoproteins, antioxidant defenses and colon function. METHODS: Twelve hyperlipidemic women followed two diets for four weeks starting with a refined food diet. Subjects then directly crossed over to the phytochemical-rich diet. Duplicate, fasting serum lipids and single, fasting antioxidant enzymes were measured at the end of the four-week refined-food diet period (baseline) and again at the end of the phytochemical-rich diet period. RESULTS: Total energy and total fat intake were similar during both diet periods, but there was a decrease in saturated fat (SFA) of 61% in the phytochemical-rich diet group. Dietary fiber, vitamin E, vitamin C and carotene intakes were 160%, 145%, 160% and 500% more, respectively, than during the refined-food diet period. The phytochemical rich diet induced a drop of 13% in total cholesterol (TC) (p < 0.05) and 16% (p < 0.001) in low density lipoprotein-cholesterol (LDL-C). Erythrocyte superoxide dismutase decreased 69% (p < 0.01) and glutathione peroxidase dropped 35% (p < 0.01). Colon function was improved on the phytochemical-rich diet. CONCLUSIONS: A diet abundant in phytochemically-rich foods beneficially affected lipoproteins, decreased need for oxidative defense mechanisms and improved colon function. PMID- 10682878 TI - Serum vitamin B12, C and folate concentrations in the New Mexico elder health survey: correlations with cognitive and affective functions. AB - OBJECTIVES: 1) To compare serum vitamin B12, C and folate concentrations in a randomly selected sample of elderly (age 65 years or older) male and female Hispanics and nonHispanic whites (NHW) and 2) to examine associations between serum B12, C and folate concentrations compared to measures of cognitive and affective (depression) functions. METHODS: Equal numbers of male and female Hispanics and NHW were randomly sampled from the Health Care Financing Administration (Medicare) registrant list for Bernalillo County, New Mexico, and asked to volunteer for a paid home interview followed by a paid comprehensive interview/examination covering health and health-related issues. In addition to serum determinations of B12, C and folate, associations were examined between these vitamins and measures of cognitive and affective functions. RESULTS: Males and Hispanics had lower serum vitamin B12, C and folate concentrations than females and NHW respectively. Participants taking a multivitamin supplement (MVI) had higher serum vitamin concentrations than those not taking MVI. There were significant associations between serum folate concentrations and measures of cognitive function, not seen with B12 or C, nor between any of the vitamins and affective function. CONCLUSIONS: Hispanics, even after adjustments for gender, age, vitamin supplementation, vitamin content of dietary foods, education and household income, had lower serum concentrations of B12, C and folate than NHW. The most significant associations observed were those between serum folate and various measures of cognitive function, even after adjusting for presence of depression. PMID- 10682879 TI - Predictors of screening for breast, cervical, colorectal, and prostatic cancer among community-based primary care practices. AB - BACKGROUND: As we enter the year 2000, it is worth looking at whether primary care practices are reaching the goals established in Healthy People 2000 for breast, cervical, colorectal, and prostatic cancer screening. The objectives of this study were (1) to determine the current rates of cancer screening; and (2) to determine which factors predict completion of a single screening test, of all tests for each cancer, and of all procedures for age and sex. METHODS: Medical records of 200 eligible patients (100 men and 100 women) from each of 24 community-based primary care practices were abstracted for cancer-screening events. RESULTS: We audited 5125 charts. A Papanicolaou smear was documented for 63.8% of women with an intact cervix within 3 years of the audit.. We found that 46.8% of women had documentation of ever having a discussion of breast self examination. For breast cancer screening, 41.8% of the women had a clinical breast examination within 1 year, 48.2% aged 40 to 49 years had a mammogram within 2 years, and 38.5% aged 50 years and older had a mammogram within 1 year. Only 29% of women aged 40 to 49 years and 17% of women 50 years and older were current for all breast cancer-screening tests. Among patients 50 years and older, 33% of men and 38% of women had a digital rectal examination within 1 year, 26% of men and 28% of women had a fecal occult blood test within 1 year, and 22% of men and 16.8% of women had a flexible sigmoidoscopy within 5 years. Of all men 28.7% had a prostate-specific antigen test within 1 year. Completion of all tests relevant for age and sex were documented for 8.6% of women aged 40 to 49 years, 3% of women 50 years and older, and 5% of men 50 years and older. The single most significant predictor of documented cancer screening was a health maintenance visit. CONCLUSIONS: This sample of primary care clinicians has not reached the goals set in Healthy People 2000 for cancer screening. Interventions aimed at increasing the percentage of patients who schedule a health maintenance visit could serve to increase cancer screening and help us reach goals set for the year 2010. PMID- 10682881 TI - Elderly deaf patients' health care experiences. AB - BACKGROUND: Approximately 10% of the US population has some degree of hearing loss, and 2 million Americans are deaf. Most medical school curricula and major textbooks characterize deafness as pathologic condition only, which is at odds with the movement to understand the Deaf population as a minority group with a unique language and cultural tradition. Physicians might therefore be unprepared to meet the needs of deaf patients effectively and sensitively. This study seeks to understand the health care experiences of elderly Deaf adults in Richmond, Va. METHODS: The authors conducted focus groups of elderly Deaf persons. Real-time voice-interpretation of the sign language communication allowed for tape recording and full transcription. The authors independently analyzed the transcripts using an editing style, and incorporated feedback on their interpretation from participants. RESULTS: Participants experienced many practical barriers to effective health care, including problems with scheduling appointments and communicating with providers. They believed that providers are ill-prepared to care for them and worried that prejudice might be a more subtle obstacle. Participants seemed resigned to these circumstances. CONCLUSIONS: The authors suggest a possible explanation for this perspective, and make specific recommendations for three levels of competency in caring for deaf patients. When the provider and the office staff provide methods to communicate with deaf patients using telephone-assisted communication, qualified interpreters, and some basic knowledge of lipreading or sign language, the care of deaf patients is greatly enhanced and the physician-patient relationship improved. PMID- 10682882 TI - Care of the elderly patient with lower extremity amputation. AB - BACKGROUND: The elderly patient with a lower extremity amputation (LEA) remains relatively common in most family medicine practices. LEA can be categorized into three major types: partial foot, transtibial amputation, and transfemoral amputation. Family physicians have not been well trained to provide care to these patients. METHODS: A literature review was performed using the key words "lower extremity amputation," "aged" and "rehabilitation." RESULTS AND CONCLUSIONS: Appropriate medical, surgical, and rehabilitative care can have a positive effect on the functional outcome for an elderly patient with a lower extremity amputation. The family physician can be instrumental in preparing the patient and family for surgery, providing psychological support, preventing and treating complications, managing comorbid illness, and assisting in rehabilitation. In addition, the family physician is primarily responsible for the daily care needs of these patients. PMID- 10682880 TI - Adolescent preventive health visits: a comparison of two invitation protocols. AB - BACKGROUND: Adolescent health care in family practice at times creates conflicting responsibilities for parents and their teenagers. In the context of a new adolescent preventive health program in a family practice setting, we compared attendance rates using two invitation protocols, the protocols differing in their emphasis on adolescent autonomy vs parental responsibility. METHODS: One hundred six teenagers in the seventh and tenth grades were invited for preventive health visits with the family nurse and physician using two protocols. Protocol 1 involved obtaining parental consent before approaching the adolescent. With protocol 2, an invitation letter and parental consent form were mailed to the teenager, while a letter of explanation was sent concurrently to the parents. In each case, the letter of invitation was followed up by a telephone call for those who did not respond. The spontaneous response rate (a positive response after receiving the letter), agreement to attend rate (a positive response after receiving the letter or being telephoned), and the attendance rate were determined according to grade, sex, and protocol. RESULTS: The spontaneous response rate was 21%, the agreement to attend rate was 75%, and the attendance rate was 44%. Attendance rates were higher for the girls compared with the boys (54% vs 35%, P = .08) and for the seventh graders compared with the tenth graders (53% vs 31%, P = .03). The spontaneous response rate was lower among the tenth graders using protocol 2 (8% vs 37.5% with protocol 1, P = .04), while the agreement to attend rate and attendance rate did not differ for the two protocols. CONCLUSIONS: Nearly one half of this population of adolescents attended preventive health visits at the family nurse's and physician's initiative. A follow-up telephone call after the initial written invitation resulted in increased participation, while approaching the teenager or parent initially did not make a difference in attendance. This pilot study shows the potential for initiating an adolescent health program in the family practice setting. PMID- 10682883 TI - Transient hyperthyroidism of hyperemesis gravidarum: a sheep in wolf's clothing. AB - BACKGROUND: Transient hyperthyroidism of hyperemesis gravidarum (THHG) is a self limiting hyperthyroidism occurring in the context of hyperemesis gravidarum. METHODS: A literature search of MEDLINE was undertaken, and a case report of a woman with THHG in pregnancy is described. RESULTS AND CONCLUSIONS: Because thyroid function tests cannot distinguish Graves disease from THHG, the diagnosis of THHG rests largely on the concurrent development of hyperemesis and hyperthyroidism and the absence of signs and symptoms of hyperthyroidism before and during pregnancy. THHG might be responsible for 40% to 70% of thyroid function abnormalities in pregnancy. Both the thyroid function abnormalities and hyperemesis are related to elevated levels of human chorionic gonadotropin. THHG resolves by 18 weeks of pregnancy without sequelae. No treatment is required. Diagnosis of THHG by the primary care provider can prevent unnecessary treatment or referral for specialty care. PMID- 10682884 TI - Late postpartum eclampsia 16 days after delivery: case report with clinical, radiologic, and pathophysiologic correlations. AB - BACKGROUND: Postpartum eclampsia is a rare, frightening, and potentially tragic complication of hypertensive pregnancies, usually developing within 48 hours of delivery. Seizures occurring days to weeks after parturition are exceedingly uncommon and require rapid, precise clinical evaluation by multiple specialists. METHODS: A case presentation of delayed postpartum eclampsia illustrates unique features of the syndrome. Extensive review of the literature highlights pathogenesis, controversies, and dilemmas surrounding this enigmatic hypertensive disorder. RESULTS AND CONCLUSIONS: A 39-year-old hypertensive patient had an uneventful full-term delivery by her family physician only to develop headache, double vision, and recurrent tonic-clonic seizures 16 days later. Initial evaluation showed severe hypertension, diplopia, hyperreflexia, proteinuria, and hyperuricemia. She was given a magnesium sulfate infusion. Magnetic resonance imaging (MRI) documented asymmetric ischemic foci within gray matter in the distribution of the posterior cerebral arteries. All symptoms, signs, and abnormal laboratory values resolved within 4 days. A follow-up MRI showed complete resolution of all cytotoxic cortical lesions. Based on human autopsy data, radiologic investigations, and animal studies, eclampsia is believed to result from explosive vasospasm, endothelial dysfunction, and cytotoxic edema of cerebral cortex. This central nervous system vasculopathy is most prominent in the posterior cerebral vasculature and is often rapidly reversible. Difficulties in differential diagnosis, typical findings on neuroimaging, and urgent management strategies are discussed. The time limit for postpartum eclampsia probably should be lengthened to 4 weeks, as indicated by our case and other clinical series. PMID- 10682885 TI - Cost-effectiveness of primary care. AB - This article is intended only as an introduction to the use of cost-effectiveness analysis in primary care. The goals are to provide a clear understanding of the difference between the cost of a treatment and its cost-effectiveness; consider what is generally a socially acceptable range for cost-effectiveness; provide some basic criteria for critically evaluating cost-effectiveness analyses in the medical literature; give some examples of the cost-effectiveness of various treatments in primary care; and provide for comparison some examples of cost effectiveness in the world of specialty care. For those interested in more detail, excellent books and reviews are available, including the report of a US. Public Health Service-appointed expert panel. PMID- 10682886 TI - Combined ipratropium and beta2--adrenergic receptor agonist in acute asthma. PMID- 10682887 TI - Rhabdomyolysis in a teenage boy: a case report. PMID- 10682888 TI - Jaundice and disseminated intravascular coagulopathy in pregnancy. PMID- 10682889 TI - Toothpaste allergy with intractable perioral rash in a 10-year old boy. PMID- 10682890 TI - Acute intermittent porphyria with seizure and paralysis in the puerperium. PMID- 10682891 TI - Health care of the deaf--toward a new understanding. PMID- 10682892 TI - Implementing clinical preventive medicine: time to fish or cut bait. PMID- 10682893 TI - Critical appraisal of the literature. PMID- 10682894 TI - Firearm safety as preventive medicine. PMID- 10682895 TI - Birth and death through a child's eyes. PMID- 10682896 TI - Birth and death through a child's eyes. PMID- 10682897 TI - Birth and death through a child's eyes. PMID- 10682898 TI - Birth and death through a child's eyes. PMID- 10682899 TI - Regulation of calcium in salivary gland secretion. AB - Neurotransmitter-regulation of fluid secretion in the salivary glands is achieved by a coordinated sequence of intracellular signaling events, including the activation of membrane receptors, generation of the intracellular second messenger, inositol 1,4,5, trisphosphate, internal Ca2+ release, and Ca2+ influx. The resulting increase in cytosolic [Ca2+] ([Ca2+]i) regulates a number of ion transporters, e.g., Ca2+-activated K+ channel, Na+/K+/2Cl- co-transporter in the basolateral membrane, and the Ca2+-activated Cl- channel in the luminal membrane, which are intricately involved in fluid secretion. Thus, regulation of [Ca2+]i is central to the regulation of salivary acinar cell function and is achieved by the concerted activities of several ion channels and Ca2+-pumps localized in various cellular membranes. Ca2+ pumps, present in the endoplasmic reticulum and the plasma membrane, serve to remove Ca2+ from the cytosol. Ca2+ channels present in the endoplasmic reticulum and the plasma membrane facilitate rapid influx of Ca2+ into the cytosol from the internal Ca2+ stores and from the external medium, respectively. It is well-established that prolonged fluid secretion is regulated via a sustained elevation in [Ca2+]i that is primarily achieved by the influx of Ca2+ into the cell from the external medium. This Ca2+ influx occurs via a putative plasma-membrane-store-operated Ca2+ channel which has not yet been identified in any non-excitable cell type. Understanding the molecular nature of this Ca2+ influx mechanism is critical to our understanding of Ca2+ signaling in salivary gland cells. This review focuses on the various active and passive Ca2+ transport mechanisms in salivary gland cells--their localization, regulation, and role in neurotransmitter-regulation of fluid secretion. In addition to a historical perspective of Ca2+ signaling, recent findings and challenging problems facing this field are highlighted. PMID- 10682900 TI - The impact of molecular genetics on oral health paradigms. AB - As a result of our increased understanding of the human genome, and the functional interrelationships of gene products with each other and with the environment, it is becoming increasingly evident that many human diseases are influenced by heritable alterations in the structure or function of genes. Significant advances in research methods and newly emerging partnerships between private and public sector interests are creating new possibilities for utilization of genetic information for the diagnosis and treatment of human diseases. The availability and application of genetic information to the understanding of normal and abnormal human growth and development are fundamentally changing the way we approach the study of human diseases. As a result, the issues and principles of medical genetics are coming to bear across all disciplines of health care. In this review, we discuss some of the potential applications of human molecular genetics for the diagnosis and treatment of oral diseases. This discussion is presented in the context of the ongoing technological advances and conceptual changes that are occurring in the field of medical genetics. To realize the promise of this new molecular genetics, we must be prepared to foresee the possibilities and to incorporate these newly emergent technologies into the evolving discipline of dentistry. By using examples of human conditions, we illustrate the broad application of this emerging technology to the study of simple as well as complex genetic diseases. Throughout this paper, we will use the following terminology: Penetrance--In a population, defined as the proportion of individuals possessing a disease-causing genotype who express the disease phenotype. When this proportion is less than 100%, the disease is said to have reduced or incomplete penetrance. Polymerase chain reaction (PCR)--A technique for amplifying a large number of copies of a specific DNA sequence flanked by two oligonucleotide primers. The DNA is alternately heated and cooled in the presence of DNA polymerase and free nucleotides, so that the specified DNA segment is denatured, hybridized with primers, and extended by DNA polymerase. MIM--Mendelian Inheritance in Man catalogue number from V. McKusick's Mendelian Inheritance in man (OMIM, 1998). PMID- 10682901 TI - Acute and chronic craniofacial pain: brainstem mechanisms of nociceptive transmission and neuroplasticity, and their clinical correlates. AB - This paper reviews the recent advances in knowledge of brainstem mechanisms related to craniofacial pain. It also draws attention to their clinical implications, and concludes with a brief overview and suggestions for future research directions. It first describes the general organizational features of the trigeminal brainstem sensory nuclear complex (VBSNC), including its input and output properties and intrinsic characteristics that are commensurate with its strategic role as the major brainstem relay of many types of somatosensory information derived from the face and mouth. The VBSNC plays a crucial role in craniofacial nociceptive transmission, as evidenced by clinical, behavioral, morphological, and electrophysiological data that have been especially derived from studies of the relay of cutaneous nociceptive afferent inputs through the subnucleus caudalis of the VBSNC. The recent literature, however, indicates that some fundamental differences exist in the processing of cutaneous vs. other craniofacial nociceptive inputs to the VBSNC, and that rostral components of the VBSNC may also play important roles in some of these processes. Modulatory mechanisms are also highlighted, including the neurochemical substrate by which nociceptive transmission in the VBSNC can be modulated. In addition, the long term consequences of peripheral injury and inflammation and, in particular, the neuroplastic changes that can be induced in the VBSNC are emphasized in view of the likely role that central sensitization, as well as peripheral sensitization, can play in acute and chronic pain. The recent findings also provide new insights into craniofacial pain behavior and are particularly relevant to many approaches currently in use for the management of pain and to the development of new diagnostic and therapeutic procedures aimed at manipulating peripheral inputs and central processes underlying nociceptive transmission and its control within the VBSNC. PMID- 10682902 TI - Biomarkers and molecular epidemiology and chemoprevention of oral carcinogenesis. AB - Chemopreventives are chemicals that prevent the formation of cancers such as oral cancer. They can take the form of nutrients or synthetic molecules, and their fundamental characteristic is that they do not produce disease processes that would result in debilitating symptoms. Current evidence indicates that they function by modifying the oxidative state of transforming cells. Biomarkers can take the form of genetic and molecular indicators, which characterize the function of chemopreventives and cancer processes such as oral carcinogenesis. Biomarkers cannot provide all the required information for risk assessment or possible activity of the chemopreventives. Other methods, such as epidemiological analyses and techniques, must be used to enhance our understanding of the risk for oral cancer in human populations. One common epidemiologic method, the questionnaire, helps to determine the use and carcinogenic potential of tobacco and alcohol during oral carcinogenesis. Genetic and molecular changes in human patient populations may result in a reduction in the number and function of tumor suppressor genes. If these changes are to be assessed, the tissues (e.g., buccal mucosa) must be accessible and harvested in a reliable and consistent manner for the acquisition of DNA, mRNA, and protein. Oral tissues provide sufficient quantities of these molecules and, under stringent conditions, the quality required for the isolation of these molecular constituents. In conjunction with epidemiologic techniques, various genotypic polymorphisms, such as glutathione-S transferase (GSTM1) or cytochrome P450 (CYP450A1), have indicated a loss in carcinogen detoxification or the processing of internal growth control signals. Biomarkers are composed of a large diverse group of genetic and molecular structures. Some of these biomarkers are indicators for programmed cell death (PCD), while others describe malignant tumor growth. Many of these classes of molecules are oxidative-responsive (e.g., tumor suppressor p53, Bcl-2, growth factors, immune-derived proteins, and death-inducing molecules) and induce PCD by triggering a cascade of cysteine proteases and regulators (e.g., caspases, death receptors). This pathway results in cell-cycle alterations and DNA fragmentation. It is hoped that a detailed knowledge of the processes involved in malignant transformation will better define the biomarker-screening tools for oral cancer. These tools will enhance our ability to predict the incidence of cancer, detect early malignant change, and quantitate chemoprevention during oral carcinogenesis. Chemopreventives such as the retinoids have already demonstrated their ability to suppress potential malignant changes in pre-malignant oral leukoplakias and decrease the incidence of second head-and-neck cancer primaries. It is our hope that this review will increase investigators' interest in developing new screening and detection systems for oral cancer. PMID- 10682903 TI - Biomechanics of the mandible. AB - In this review the biomechanical behavior of the mandibular bone tissue, and of the mandibular bone as a whole, in response to external loading is discussed. A survey is given of the determinants of mandibular stiffness and strength, including the mechanical properties and distribution of bone tissue and the size and shape of the mandible. Mandibular deformations, stresses, and strains that occur during static biting and chewing are reviewed. During biting and the powerstroke of mastication, a combination of sagittal bending, corpus rotation, and transverse bending occurs. The result is a complex pattern of stresses and strains (compressive, tensile, shear, torsional) in the mandible. To be able to resist forces and bending and torsional moments, not only the material properties of the mandible but also its geometrical design is of importance. This is reflected by variables like polar and maximum and minimum moments of inertia and the relative amount and distribution of bone tissue. In the longitudinal direction, the mandible is stiffer than in transverse directions, and the vertical cross-sectional dimension of the mandible is larger than its transverse dimension. These features enhance the resistance of the mandible to the relatively large vertical shear forces and bending moments that come into play in the sagittal plane. PMID- 10682904 TI - Commentary on the challenges of establishing gene medicines as a new class of therapeutics. PMID- 10682905 TI - Non-viral peptide-based approaches to gene delivery. AB - To achieve effective non-viral gene therapy, the control of in vitro and in vivo stability, cellular access, intracellular trafficking and nuclear retention of plasmids must be achieved. Inefficient endosomal release, stability against cytosolic nucleases, cytoplasmic transport and nuclear entry of plasmids are amongst some of the key limiting factors in the use of plasmids for effective gene therapy. Synthetic peptide-based gene delivery systems can be designed for DNA compaction, serum stability, cell-specific targeting, endosomolysis, cytoplasmic stability and nuclear transport. The stability of compacted DNA under physiological conditions can be enhanced by the use of hydrophilic polymers, such as polyethylene glycol. The aims of this review are to (i) explore theoretical and experimental aspects of DNA compaction, (ii) describe approaches for stabilizing compacted DNA, (iii) assess techniques used for characterization of compacted DNA, and (iv) review possible use of peptides for efficient gene transfer. PMID- 10682906 TI - Gene transfer into the CNS using recombinant adeno-associated virus: analysis of vector DNA forms resulting in sustained expression. AB - Recombinant adeno-associated virus (rAAV) vectors have shown significant promise as vehicles for in vivo gene transfer, particularly for transduction of organs composed primarily of non-dividing cells (i.e., muscle, CNS, and liver). However, the mechanistic basis for this desirable property remains unclear. To investigate the fate of rAAV genomes in mouse brain, we stereotactically injected an rAAV vector carrying the E. coli lacZ gene into the caudate of BALB/c mice and demonstrate efficient transduction of mouse brain cells that possess cellular morphology consistent with post-mitotic neurons. We observed a significant increase in beta-galactosidase expression from 5 to 56 days after injection that paralleled the disappearance of single-stranded DNA input genomes. Analysis of in vivo viral DNA forms over time out to 5 months after inoculation revealed that rAAV genomes associated with high molecular weight mouse chromosomal DNA by 14 days after injection and persisted for the length of this study. The pattern of Southern hybridization was consistent with random viral integration in predominantly head-to-tail concatameric arrays. Importantly, we also documented an additional DNA species that appears to be a monomeric episomal circular form based on nuclease sensitivity assays. These data are the first to document the existence of multiple vector DNA forms present within the adult murine brain following direct rAAV inoculation and therefore, provide insight into the molecular events that ultimately result in long-term rAAV mediated transgene expression. PMID- 10682907 TI - Synthesis and characterization of aromatic ring-based cationic lipids for gene delivery in vitro and in vivo. AB - A new series of cationic lipids has been synthesized for gene delivery using 3,5 dihydroxybenzyl alcohol as the backbone and starting material. Using CMV driven expression system and luciferase gene as a reporter, we demonstrated that the transfection activity of these new lipids when formulated with Tween 80 as co lipid is comparable to that of DOTAP, one of the most commonly used cationic lipids for transfection. Among the four different cell lines tested including murine melanoma BL-6 cells, human embryonic kidney 293 cells, HepG2 and HeLa cells, the highest transgene expression was seen in 293 cells. Results from in vivo experiments using mice as an animal model show that these cationic lipids preferentially transfect the cells in the lung upon tail vein administration. The cationic lipid, N,N,N-trimethyl-N-[3,5-bis(tetradecyloxy)benzyl] ammonium bromide 4c(di-C14:0) with two 14-hydrocarbon chains exhibits the best transfection activity. These results suggest that these new aromatic ring-based cationic lipids are useful transfection reagents for both in vitro and in vivo gene transfer studies. PMID- 10682908 TI - Development of transferrin-polycation/DNA based vectors for gene delivery to melanoma cells. AB - We describe the comparison of non-viral polycation transfection reagents, adenovirus-enhanced transferrinfection (AVET), polyethylenimine (PEI800) and transferrin-conjugated PEI800 (Tf-PEI800) in their ability to transfect murine and primary human melanoma cell lines. Expression of a reporter gene, cell surface marker and secreted protein (interleukin-2) was assessed for each vector system. Testing for luciferase reporter gene expression in murine and primary human cell lines, AVET and Tf-PEI800, both showed high levels of expression and comparable activity. Furthermore, when the melanoma cell line B16F10 was transfected with a cell surface marker up to approximately 97% of the cells expressed the protein on the cell surface. Assessing the levels of secreted IL-2 in murine cell lines, AVET/IL-2, Tf-PEI800/IL-2 and PEI800/IL-2 all expressed high levels of the cytokine (up to 20 microg IL-2/10(6) cells/24 h). In primary human melanoma cell lines, AVET/IL-2 transfected cells secreted more IL-2 than cells transfected with either Tf-PEI800/IL-2 or PEI800/IL-2. In murine melanoma cell culture experiments, positively charged PEI800/DNA and Tf-PEI800/DNA complexes gave similar transfection efficiencies. However, when subcutaneous tumors in mice were injected with the luciferase reporter gene complexed with either Tf-PEI800 or AVET, higher transfection activity was measured in the tumors as compared to ligand free PEI800/DNA complexes. PMID- 10682909 TI - Optimisation of polyethylenimine-based gene delivery to mouse brain. AB - Polyethylenimine (PEI) is proving to be an efficient and versatile vector for gene delivery in vivo. However, a limiting factor is the relatively short duration of gene expression in some sites. Given the particularly high levels of expression seen in the short term we postulated that loss of expression could result from overloading the nucleus with foreign DNA. To address this problem we first followed DNA delivery and localisation with digoxin-labelled plasmid DNA complexed with 22 kD linear PEI and used these complexes for intraventricular injection into brains of anaesthetised newborn mice. At 24 h post-injection, labelled DNA was found exclusively in the nuclear and perinuclear regions. We next carried out a dose response curve using decreasing amounts of DNA, either in a constant volume (2 microl) or at a constant concentration (500 ng/microl). In both conditions, transgene expression yield was maximum at 100 ng DNA per injection. Using this optimal amount of DNA increased yield of transgene expression significantly at 24 h and one week post-injection as compared to 1 microg DNA. A final point addressed was whether co-expressing an anti-apoptotic gene could enhance gene expression in the longer term. Co-expressing bcl-X(L) with luciferase or LacZ significantly increased expression of both these genes at one week post-injection. PMID- 10682910 TI - Comparison of process parameters for microencapsulation of plasmid DNA in poly(D,L-lactic-co-glycolic) acid microspheres. AB - Poly(D,L-lactic-co-glycolic) acid (PLGA) microspheres containing plasmid DNA encoding the firefly luciferase gene were prepared using the water-in-oil-in water (w/o/w) double emulsion and solvent evaporation method. In this study, we investigated the effects of three process parameters on DNA microencapsulation: (1) emulsification method used to generate the primary emulsion, (2) water/oil ratio during formation of the first emulsion, and (3) surfactant concentration used in the preparation of the second emulsion. The resulting formulations were also analyzed for microsphere size, encapsulation efficiency, and kinetics of DNA release. We found that although each process alteration resulted in encapsulation of biologically active, structurally intact DNA, the surfactant and water/oil ratio significantly affected the size, release kinetics and encapsulation efficiency of plasmid DNA. PMID- 10682911 TI - Inactivation of the reconstituted oxoglutarate carrier from bovine heart mitochondria by pyridoxal 5'-phosphate. AB - The effect of pyridoxal 5'-phosphate and some other lysine reagents on the purified, reconstituted mitochondrial oxoglutarate transport protein has been investigated. The inhibition of oxoglutarate/oxoglutarate exchange by pyridoxal 5'-phosphate can be reversed by passing the proteoliposomes through a Sephadex column but the reduction of the Schiff's base by sodium borohydride yielded an irreversible inactivation of the oxoglutarate carrier protein. Pyridoxal 5' phosphate, which caused a time- and concentration-dependent inactivation of oxoglutarate transport with an IC50 of 0.5 mM, competed with the substrate for binding to the oxoglutarate carrier (Ki = 0.4 mM). Kinetic analysis of oxoglutarate transport inhibition by pyridoxal 5'-phosphate indicated that modification of a single amino acid residue/carrier molecule was sufficient for complete inhibition of oxoglutarate transport. After reduction with sodium borohydride [3H]pyridoxal 5'-phosphate bound covalently to the oxoglutarate carrier. Incubation of the proteoliposomes with oxoglutarate or L-malate protected the carrier against inactivation and no radioactivity was found associated with the carrier protein. In contrast, glutarate and substrates of other mitochondrial carrier proteins were unable to protect the carrier. Mersalyl, which is a known sulfhydryl reagent, also failed to protect the oxoglutarate carrier against inhibition by pyridoxal 5'-phosphate. These results indicate that pyridoxal 5'-phosphate interacts with the oxoglutarate carrier at a site(s) (i.e., a lysine residue(s) and/or the amino-terminal glycine residue) which is essential for substrate translocation and may be localized at or near the substrate-binding site. PMID- 10682912 TI - Oligomeric state of wild-type and cysteine-less yeast mitochondrial citrate transport proteins. AB - Experiments have been conducted to determine the oligomeric state of the mitochondrial citrate transport protein (CTP) from the yeast Saccharomyces cerevisiae. Both wild-type and cysteine-less (Cys-less) CTPs were overexpressed in E. coli and solubilized with sarkosyl. The purity of the solubilized material is approximately 75%. Upon incorporation into phospholipid vesicles, a high specific transport activity is obtained with both the wild-type and Cys-less CTPs, thereby demonstrating the structural and functional integrity of the preparations. Two independent approaches were utilized to determine native molecular weight. First, CTP molecular weight was determined via nondenaturing size-exclusion chromatography. With this methodology we obtained molecular weight values of 70,961 and 70,118 for the wild-type and Cys-less CTPs, respectively. Second, charge-shift native gel electrophoresis was carried out utilizing a low concentration of the negatively charged detergent sarkosyl, which served to both impart a charge shift to the CTP and the protein standards, as well as to promote protein solubility. Via the second method, we obtained molecular weight values of 69,122 and 74,911 for the wild-type and Cys-less CTPs, respectively. Both methods clearly indicate that following solubilization, the wild-type and the Cys-less CTPs exist exclusively as dimers. Furthermore, disulfide bonds are not required for either dimer formation or stabilization. The dimeric state of the CTP has important implications for the structural basis underlying the CTP translocation mechanism. PMID- 10682913 TI - Inhibitory properties of ruthenium amine complexes on mitochondrial calcium uptake. AB - The recent finding that the inhibition of Ca2+-stimulated respiration by ruthenium red is mainly due to a binuclear ruthenium complex (Ru360) present in the commercial samples of the classical inhibitor ruthenium red (Ying et. al., 1991), showed that this complex is the more potent and specific inhibitor of the mitochondrial calcium uniporter. This work was aimed to provide insights into the mechanism by which Ru360 and other ruthenium-related compounds inhibits calcium uptake. Ruthenium red and a synthesized analog (Rrphen) were compared with Ru360. The inhibition by this binuclear complex was noncompetitive, with a Ki of 9.89 nM. The number of specific binding sites for Ru360 was 6.2 pmol/mg protein. Ruthenium red and Ru360 were mutually exclusive inhibitors. Bound La3+ was not displaced by Ru360. Rrphen was the least effective for inhibiting calcium uptake. The results support the notion of a specific binding site in the uniporter for the polycationic complexes and a negative charged region from the phospholipids in the membrane, closely associated with the uniporter inhibitor-binding site. PMID- 10682915 TI - Binding of rat brain hexokinase to recombinant yeast mitochondria: identification of necessary molecular determinants. AB - The association in vitro of rat brain hexokinase to mitochondria from rat liver or yeast (wild type, porinless, or expressing recombinant human porin) was studied in an effort to identify minimal requirements for each component. A short hydrophobic N-terminal peptide of hexokinase, readily cleavable by proteases, is absolutely required for its binding to all mitochondria. Mammalian porins are significantly cleaved at two positions in putative cytoplasmic loops around residues 110 and 200, as determined by proteolytic-fragment identification using antibodies. Recombinant human porin in yeast mitochondria is more sensitive to proteolysis than wild-type porin in rat liver mitochondria. Recombinant yeast mitochondria, harboring several natural or engineered porins from various sources, bind hexokinase to variable extent with marked preference for the mammalian porin1 isoform. Genetic alteration of this isoform at the C-, but not the N-terminal, results in a significant reduction of hexokinase binding ability. Macromolecular crowding (dextran) promotes a stronger association of the enzyme to all recombinant mitochondria, as well as to proteolytically digested organelles. Consequently, brain hexokinase association with heterologous mitochondria (yeast) in these conditions occurs to an extent comparable to that with homologous (rat) mitochondria. The study, also pertinent to the topology and organization of porin in the membrane, represents a necessary first step in the functional investigation of the physiological role of mammalian hexokinase binding to mitochondria in reconstituted heterologous recombinant systems, as models to cellular metabolism. PMID- 10682914 TI - Stimulation of mitochondrial gene expression and proliferation of mitochondria following impairment of cellular energy transfer by inhibition of the phosphocreatine circuit in rat hearts. AB - Mitochondria proliferate when cellular energy demand increases. However, the pathways leading to enhanced expression of mitochondrial genes are largely unknown. We tested the hypothesis that an altered flux through energy metabolism is the key regulatory event by decreasing mitochondrial energy supply to rat heart cells by creatine depletion. Electron microscopy showed that the density of mitochondria increased by 75% in such hearts (p < 0.01). Levels of representative mRNAs encoded on mitochondrial DNA (mtDNA) or on nuclear chromosomes were elevated 1.5 to 2-fold (p < 0.05), while the mtDNA content was unchanged. The mRNA for the nuclear encoded mitochondrial transcription factor A (mtTFA) was increased after GPA feeding (p < 0.05). Thus, we have shown that an impairment of mitochondrial energy supply causes stimulation of gene expression resulting in mitochondrial proliferation, probably as a compensatory mechanism. The observed activation of the mtTFA gene corroborates the important function of this protein in nuclear-mitochondrial communication. PMID- 10682916 TI - Mitochondrial effects of triarylmethane dyes. AB - The mitochondrial effects of submicromolar concentrations of six triarylmethane dyes, with potential applications in antioncotic photodynamic therapy, were studied. All dyes promoted an inhibition of glutamate or succinate-supported respiration in uncoupled mitochondria, in a manner stimulated photodynamically. No inhibition of N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) supported respiration was observed, indicating that these dyes do not affect mitochondrial complex IV. When mitochondria were energized with TMPD in the absence of an uncoupler, treatment with victoria blue R, B, or BO, promoted a dissipation of mitochondrial membrane potential and increase of respiratory rates, compatible with mitochondrial uncoupling. This effect was observed even in the dark, and was not prevented by EGTA, Mg2+ or cyclosporin A, suggesting that it is promoted by a direct effect of the dye on inner mitochondrial membrane permeability to protons. Indeed, victoria blue R, B, and BO promoted swelling of valinomycin-treated mitochondria incubated in a hyposmotic K+-acetate-based medium, confirming that these dyes act as classic protonophores such as FCCP. On the other hand, ethyl violet, crystal violet, and malachite green promoted a dissipation of mitochondrial membrane potential, accompanied by mitochondrial swelling, which was prevented by EGTA, Mg2+, and cyclosporin A, demonstrating that these drugs induce mitochondrial permeability transition. This mitochondrial permeabilization was followed by respiratory inhibition, attributable to cytochrome c release, and was caused by the oxidation of NAD(P)H promoted by these drugs. PMID- 10682917 TI - Myopathic mutations affect differently the inactivation of the two gating modes of sodium channels. AB - Three groups of mutations of the alpha subunit of the rat skeletal muscle sodium channel (rSkM1), homologous to mutations linked to human muscle hereditary diseases, have been studied by heterologous expression in frog oocytes: S798F, G1299E, G1299V, and G1299A, linked with potassium-aggravated myotonia (PAM); T1306M, R1441C and R1441P, linked with paramyotonia congenita (PC); T698M and M1353V, linked with the hyperkalemic periodic paralysis (HyPP). Wild-type rSkM1 channels (WT) show two gating modes, M1 and M2, which differ mainly in the process of inactivation. The naturally most representative mode M1 is tenfold faster and develops at approximately 30 mV less depolarized potentials. A common feature of myopathy-linked mutants is an increase in the mode M2 probability, P(M2), but phenotype-specific alterations of voltage-dependence and kinetics of inactivation of both modes are also observed. The coexpression of the sodium channel beta1 subunit, which has been studied for WT and for the five best expressing mutants, generally caused a threefold reduction of P(M2) without changing the properties of the individual modes. This indicates that the mutations do not affect the alpha - beta1 interaction and that the phenotypic changes in P(M2) observed for the enhanced mode M2 behavior of the sole alpha subunits, although largely depressed in the native tissue, are likely to be the most important functional modification that causes the muscle hyperexcitability observed in all patients carrying the myotonic mutations. The interpretation of the more phenotype-specific changes revealed by our study is not obvious, but it may offer clues for understanding the different clinical manifestations of the diseases associated with the various mutations. PMID- 10682918 TI - Role of pinoline and melatonin in stabilizing hepatic microsomal membranes against oxidative stress. AB - We investigated the influence of pinoline (0.01-1.5 mM) on microsomal membrane fluidity before and after rigidity was induced by oxidative stress. In addition, we tested the effect of pinoline in the presence of 1 mM melatonin. The fluidity in rat hepatic microsomes was monitored using fluorescence spectroscopy and it was compared to the inhibition of malonaldehyde (MDA) plus 4-hydroxyalkenals (4 HDA) production as a reflection of lipid peroxidation. Below 0.6 mM, pinoline inhibited membrane rigidity in a manner parallel to its inhibitory effect on MDA + 4-HDA formation. At concentrations between 1-1.5 mM, pinoline was less effective in stabilizing microsomal membranes than was predicted from its inhibition of lipid peroxidation. The addition of 1 mM melatonin enhanced the membrane-stabilizing activity of pinoline (0.01-0.6 mM). This cooperative effect was not observed for concentrations of pinoline between 1-1.5 mM. When pinoline was tested without induced oxidative damage, 1-1.5 mM pinoline maintained membrane fluidity at the same level as that recorded after induced lipid peroxidation. The results suggest that pinoline may be another pineal molecule that prevents membrane rigidity mediated by lipid peroxidation and this ability is enhanced by melatonin. PMID- 10682919 TI - A quantitative approach to the evaluation of the morphological variability of two echinostomes, Echinostoma miyagawai Ishii, 1932 and E. revolutum (Frolich, 1802), from Europe. AB - A comparative morphometric analysis was conducted on two European species of Echinostoma in order to examine the degree of the variability in the metrical characteristics of the adults and to assess their value in discriminating species. Adult E. miyagawai and E. revolutum, obtained experimentally, were compared using univariate and multivariate statistical analyses of 35 and 25 metrical characters, respectively. All subsets of worms of different ages represented homogeneous samples with respect to their morphometric characteristics; however, univariate analyses revealed significant differences in 22 and 23 variables between the corresponding age subsets of the two species, and it was found that the different allometric growth patterns contribute to this. The variables, body width at the posterior border of the ventral sucker, pharynx length and width, ovary length, testes length and width and length of the pre ovarian region, exhibited isometric or positive allometric growth in E. miyagawai and negative allometry in E. revolutum. A cluster analysis based on 61 specimens and 25 variables separated E. revolutum and E. miyagawai unambiguously, producing an exact ordering of the specimens with respect to their identity and age. A forward stepwise discriminant analysis identified five variables (body width at the posterior border of ventral sucker, head collar width, length of oesophagus, width of ventral sucker and length of the pre-ovarian region) which yielded a 100% accurate classification of the two species. We suggest, therefore, that the morphometric characteristics of the adult worms should be used in studies attempting the identification of species or isolates of Echinostoma spp. More comparative data need to be gathered in order that the species boundaries within the 'revolutum' group be defined more accurately. PMID- 10682920 TI - Observations on trichodinid ectoparasites (Ciliophora: Peritricha) from the gills of maricultured molluscs in China, with descriptions of three new species of Trichodina Ehrenberg, 1838. AB - During surveys of the trichodinid parasites in mariculture beds off the coast of Shandong Province, China, four species of the genus Trichodina from the gills of marine molluscs were investigated and morphologically studied. Of these, three are described as new: T. ruditapicis n. sp. from Ruditapes philippinarum (Veneridae), T. scapharcae n. sp. from Scapharca subcrenata (Arcidae) and T. mactrae n. sp. from Mactra veneriformis (Mactridae). One little-known species, T. macomarum Raabe & Raabe, 1959, is redescribed from M. veneriformis. Taxonomic and morphometric data for these trichodinids based on wet silver nitrate and protargol-impregnated specimens are presented. For each of the new species, comparisons with closely related species are provided. PMID- 10682921 TI - Durettechina beveridgei n. g., n. sp. (Nematoda: Seuratidae) from Antechinus spp. (Dasyuridae: Marsupialia) from Australia. AB - Durettechina beveridgei n. g., n. sp. (Nematoda: Seuratidae) is described from Antechinus flavipes (Dasyuridae) from Victoria and New South Wales. A single female from A. bellus from the Northern Territory may also be D. beveridgei. This new genus is compared with other genera of the Echinonematinae, to which it has been assigned. The genus has a unique body armature and most closely resembles Chabaudechina, in the armature of the cephalic bulb, but has four rather than five rows of hooks, and Linstowinema, in having body hooks on the cuticle of the anterior region, but has 18-22 hooks in each row rather than 14-16. The hooks of Durettechinca are also smaller and have a less complex root morphology than those of Linstowinemna. Durettechina resembles Seurechinac and Chabaudechina in having caudal alae into which papillae extend, but differs from both these genera in the number and arrangement of the caudal papillae, as well as in the body armature. Durettechina, is most different from Bainechina, which has neither hooks on a cephalic bulb nor body hooks on the anterior region nor caudal alae. PMID- 10682922 TI - Six new species of Lepidapedon Stafford, 1904 (Digenea: Lepocreadiidae) from deep sea macrourid fishes from the Gulf of Mexico and Caribbean Sea, with revised keys to the species of the genus. AB - Species of the genus Lepidapecon are divided into various groups and subgroups based on vitelline distribution relative to the acetabulum and anterior extent of the excretory vesicle. Members of this genus predominantly parasitise gadiform fishes and are commonly collected from relatively deep waters. A recent study of deep-sea helminths from macrourids of the Gulf of Mexico and Caribbean Sea revealed six new species of this genus. L. mexicanensis n. sp., of the elongatum group, elongatum subgroup, differs from other species in this subgroup in proportions (as % of body length), lacking confluent vitelline fields between both the ovary and anterior testis and the testes, and in having a smaller egg and body size. L. nezumiatis n. sp., of the elongatum group, desclersae oesophagus than prepharynx. L. caribbaei n. sp. and L. longivesicula n. sp., of the garrardi group, congeri sub-subgroup, differs from both L. filiformis and L. desclersae in having intermediate egg and body sizes, and a longer group, differ from L. congeri in having a sucker-ratio of 1: < 1. L. caribbaei n. sp. and L. longivesicula n. sp. differ from each other in that L. caribbaei n. sp. has numerous long, barb-like, deeply imbedded spines, a less elongate body, an infundibuliform oral sucker, a similar-sized oesophagus and prepharynx, and a caecal bifurcation which is closer to the acetabulum than oral sucker, while L. longivesicula n. sp. has shorter, serrate or plate-like, lightly imbedded, widely to sporadically spaced spines, a more elongate body, a spherical to subspherical oral sucker, a longer oesophagus than prepharynx, and a caecal bifurcation which is closer to the oral sucker than acetabulum. L. desotoensis n. sp., of the rachion group, rachion subgroup, is distinct from both L. luteum and L. abyssensis in having a smaller size, lacking cervical glands or pharyngeal gland cells, and possessing dark-staining particles in the mesenchyme, while it differs from L. abyssensis specifically in having a much longer oesophagus than prepharynx, lateral vitelline fields that are not confluent intertesticularly, and wider eggs. L. zaniophori n. sp., also of the rachion subgroup, differs from both L. cascadensis and L. genge in having a smaller egg size, a shorter prepharynx and oesophagus than pharynx, and vitelline fields that are intertesticular but only slightly encroach between the ovary and anterior testis. L. sammari and L. spiniferi are designated as incertae sedis, and L. quiloni and L. stromateusi are designated as species inquirendae. New parasite keys and host records for Coelorinchus coelorhincus. C. caribbaeus and Nezumia cyrano are offered. Support is given to Lepidapedon probably being the dominant digenean genus in deep water. PMID- 10682923 TI - Records of the bird capillariid nematode Ornithocapillaria appendiculata (Freitas, 1933) n. comb. from freshwater fishes in Mexico, with remarks on Capillaria patzcuarensis Osorio-Sarabia et al., 1986. AB - Re-examination of capillariid specimens collected from the freshwater fish Chirostoma estor Jordan from Lake Patzcuaro in 1985-1986 and deposited as paratypes of Capillaria patzcuarensis Osorio-Sarabia, Perez-Ponce de Leon & Salgado-Maldonado, 1986 showed that their morphology was in contradiction with the description of this species and, in fact, they could be identified as the species originally described as C. appendiculata Freitas, 1933 from cormorants Phalacrocorax brasilianus (Gm.) in Brazil; conspecific capillariid specimens were later recorded from Chirostoma estor and Cyprinus carpio L. from the same locality. This species and two others are transferred to Ornithocapillaria Barus & Sergeeva, 1990 as O. appendiculata (Freitas, 1933) n. comb., O. carbonis (Dubinin & Dubinina, 1940) n. comb., and O. phalacrocoraxi (Borgarenko, 1975) n. comb. This is the first record of O. appendiculata in Mexico. Its occurrence in fishes suggests that these nematodes may be acquired by their fish hosts accidentally while feeding on cormorant excrement containing mature nematodes. A female capillariid collected from one of 110 Chirostoma estor examined from this locality in April, 1998 was identified as Capillaria patzcuarensis. Both capillariid species are briefly described and illustrated. PMID- 10682924 TI - Three new species of Polyclithrum Rogers, 1967 (Gyrodactylidae: Monogenea) from mugilid fishes from Australia and Brazil, with a redescription of P. mugilini Rogers, 1967. AB - Polyclithrum mugilini Rogers, 1967, a parasite of Mugil cephalus Linnaeus, is redescribed from type-material from Lake Seminole, Georgia, USA. Three new species of Polyclithrum Rogers, 1967 are also described: P. alberti n. sp. from M. cephalus from the Albert River, Queensland, Australia; P. boegeri n. sp. from M. platanus Gunther from Rio da Guarda, State of Rio de Janeiro, Brazil; and P. corallense n. sp. from M. cephalus from Heron Island, Great Barrier Reef, Australia. The four species can be distinguished by the size and shape of haptoral sclerites, but in particular by accessory bar number 3, the dorsal bar, the marginal hooks and the hamulus point to shaft angle. The validity of Micropolyclithrum parvum Skinner, 1975, a parasite of M. cephalus in Biscayne Bay, Florida, is discussed, and a key to the species of Polyclithrum is presented. PMID- 10682925 TI - A new species and a new record of ectoparasitic mites from thrips in Turkey (Acari: Trombidiidae and Erythraeidae). AB - Trombidium telletxeae n. sp. is described from a larva parasitic on Odontothrips sp. (Thysanoptera: Thripidae) from Turkey. Grandjeanella multisetosa Zhang & Goldarazena is also reported for the first time in Turkey. A host list for ectoparasitic larval mites on thrips in Turkey is presented. PMID- 10682926 TI - Accuracy of stereotactic coordinate transformation using a localisation frame and computed tomographic imaging. Part I. Influence of the mathematical and physical properties of the CT on the image of the rods of the localisation frame and the determination of their centres. AB - The accuracy of coordinate transformation from the computed tomographic (CT) space to the stereotactic frame space was analysed for frame-based stereotactic systems which use a localisation frame and coordinate transformation based on matrix calculation. The coordinate transformation was divided into three consecutive steps: (1) transforming the localisation frame into the CT image built up from pixels with distinct attenuation values, (2) determining the rod centres of the localisation frame in the CT image, and (3) coordinate transformation from the image to the frame space using the centres of the rods in the image space and algebraic, matrix-based calculation. The error contribution at each step was evaluated separately and its effect on the subsequent mathematical operations was analysed. The first step dealt with the influences of the mathematical and physical properties of the CT on the image of the localisation frame. Noise, slice thickness, convolution filter, dimension of the pixel matrix, and image processing had an influence on the attenuation values in each pixel. Above all, the slice thickness had an effect on the shape of the oblique rods in the CT image. At the second step, the main error contribution was due to the method by which the centre of the rods was calculated. The most accurate method was to determine the centre of gravity using the attenuation values as single mass points (with accuracy in the range of +/-1/10 pixel, or +/ 0.125 mm), followed by rounding off the centre of gravity and the highest pixel value in the square matrix R2(N) within 1 pixel. Pointing with a cursor under visual control was accurate to 1 pixel and the pixel with the highest attenuation value showed deviations of up to 2 pixels in the x and y axes. Thus, the methods differed by a factor of 20. The influence of the CT mathematics and physics on the determination of the centre of the fiducials was negligible in comparison to the method of calculation used. There was no systemic error due to the filtred back projection algorithm. Data input errors due to noise were in the range of 1/10 pixel. The effects of the remaining physical influences were all in the range of the error due to noise. In particular these results speak in favour of no influence of slice thickness on coordinate transformation. PMID- 10682927 TI - Accuracy of stereotactic coordinate transformation using a localisation frame and computed tomographic imaging. Part II. Analysis of matrix-based coordinate transformation. AB - The accuracy of coordinate transformation from a CT image to a stereotactic frame was investigated for stereotactic systems using a localisation frame and matrix based coordinate transformation. The main source of error influencing calculation was input data, due to inaccurate calculation of the centres of the rods of the localisation frame in the CT image, and the propagation of this input error during subsequent matrix calculation. Systemic errors during matrix calculation do not exist, and rounding off errors were of subordinate importance compared to the input data error. The influence of input data error on coordinate transformation was studied by geometric methods, computer simulation, and numerical analysis. In the geometric model, input data errors affected the calculation of the centres of the three oblique rods in the frame space and shifted them three points upwards or downwards on the axis of each rod. The three centres of the oblique rods defined the "CT plane" in the 3D space of the stereotactic frame. Displacements of these three centres caused a characteristic tilting of the CT plane. The positions of the correct and tilted CT planes defined the spatial error properties for all target points on the CT plane. The computer simulation investigated the effects on matrix-based transformation of all possible displacement combinations on the three oblique rods by 1 pixel (1. 16 mm) in the x and y directions. A characteristic, space-dependent distribution of the frame-related coordinates was obtained for each target point. In the centre of the frame, we found a maximal deviation of 1.0 mm in the xy direction and 2 mm in the z direction. This corresponded to an error amplification of 0.73 in the xy direction and 1.22 in the z direction relative to the error at the centres of the rods. The maximum deviation (found in the periphery) for all combinations on the three oblique rods was 1.7 mm in the xy direction and 3.3 mm in the z direction. This resulted in an amplification of 1.03 in the xy direction and 2.01 in the z direction. This results had to be multiplied by 2 to obtain a maximal error estimate for displacements including all nine rods of the localisation frame. Numerical analysis showed stable solutions with low error amplification for hexagonal frame arrangements. PMID- 10682928 TI - Expression of BCL-2 oncoprotein on tumor cells and tumor-infiltrating lymphocytes (TIL) in meningiomas. AB - The Expression of the antiapoptotic oncoprotein BCL-2 and its correlation to tumor grade in 62 meningiomas (48 classic, 9 atypical, and 5 anaplastic) using single and double immunohistochemistry was investigated. BCL-2 expression was found in two different cell populations identified as lymphocytes (BCL-2+CD3+) and tumor cells (BCL+/CD3-). Tumor-infiltrating lymphocytes (TIL) (CD3+) were found within classic (9.5% of cells), atypical (2.4% of cells), and anaplastic (1.8% of cells) meningiomas. In classic meningiomas, 66.5% of TIL were BCL-2 positive, in atypical meningiomas 79.2%, and in anaplastic meningiomas 37.9%. In 33 (68.8%) of the classic meningiomas, medium to high counts of BCL-2+ tumor cells were detected. Atypical meningiomas showed nearly equal percentages of high (two patients), medium (five patients), and low (two patients) BCL-2+ tumor cell counts, whereas anaplastic meningiomas showed only medium (two patients) and low (three patients) BCL-2 tumor cell counts or were BCL-2-negative (one patient). In summary, a significant inverse correlation between the number of BCL-2-positive tumor cells and tumor grade in meningiomas was found. These findings support the hypothesis of cell survival prolongation by the antiapoptotic ability of BCL-2 proto-oncogenes and demonstrate the prognostic relevance of BCL-2 immunoreactivity in meningiomas. PMID- 10682929 TI - Evaluation of the malignancy of glioma using 11C-methionine positron emission tomography and proliferating cell nuclear antigen staining. AB - 11C-methionine positron emission tomography (PET) and proliferating cell nuclear antigen (PCNA) staining were performed in 13 cases of glioma to investigate the relationship between the uptake of L-[methyl]-11C-methionine and the degree of malignancy and proliferative potential. The 11C-methionine uptake was significantly greater in high-grade gliomas compared to low-grade gliomas (P<0.05). The PCNA indexes were also significantly higher in the high-grade cases (P<0.05). Moreover, a strong positive correlation was found between the 11C methionine values and the PCNA indexes (P<0.005), demonstrating that higher 11C methionine uptake was associated with greater proliferative potential and greater malignancy. 11C-methionine PET is a potentially useful preoperative method to discriminate the malignancy of glioma. PMID- 10682930 TI - Fibroxanthoma arising from the cranial dura mater. AB - A primary xanthomatous tumor is very rare in the central nervous system (CNS). Here we report the case of a fibroxanthoma arising from the dura mater of the cerebrum that demonstrated no systemic disease or metabolic abnormalities. A 19 month-old, otherwise healthy boy was found to have an enlarged head. Magnetic resonance imaging demonstrated a left occipital dural mass lesion and an enlarged left cerebral hemisphere with ipsilateral ventricular enlargement. Subtotal removal of the tumor was performed through the left parieto-occipital craniotomy. The tumor was composed of a central fibrous portion, a peripheral xanthomatous area, and a boundary. The peripheral area of the tumor showed abundant uniform xanthomatous cells with a thin fibrous stroma and the mass was diagnosed as fibroxanthoma involving the dura. This may represent a distinct category of tumor, which is different from the previously reported cases of fibrous xanthoma and fibrous histiocytoma. Intracranial xanthomatous tumors may be heterogeneous in their origin and histological features. However, further studies are needed to elucidate their clinical features, biological behavior, and optimal treatment strategies. PMID- 10682931 TI - Endovascular treatment of basilar trunk aneurysm associated with fenestration of the basilar artery. AB - Basilar trunk saccular aneurysms associated with fenestration are infrequent, especially in the middle or distal portion of the basilar artery. Surgical treatment of the basilar trunk aneurysm is difficult, due to its anatomical environment and the complicated surgical exposure. A 46-year-old woman presenting with Hunt and Kosnik grade II subarachnoid hemorrhage was found to have a ruptured aneurysm arising at the proximal corner of the associated fenestration in the middle portion of the basilar artery. Because of surgical difficulties anticipated in approaching the aneurysm, it was decided to treat it with endovascular embolization utilizing the Guglielmi detachable coil; and complete occlusion of the aneurysm was obtained. The efficacy of endovascular treatment for the basilar trunk aneurysm with associated fenestration is discussed from anatomical and embryological points of view, and relevant literature is reviewed. PMID- 10682932 TI - Sphenoidal sinus mucocele after transsphenoidal surgery for acromegaly. AB - This report concerns one case of a sphenoid sinus mucocele occurring 17 years after transsphenoidal surgery for acromegaly. In 1979, a 51-year-old man was successfully operated by the transnasal transsphenoidal approach for a growth hormone (GH) adenoma 1 cm in diameter. In 1996, the patient was hospitalized for headaches and diplopia. He presented a loss of right visual acuity with paralysis of the right oculomotor nerve. The basal GH level was normal with a satisfactory decrease after oral glucose ingestion. Pituitary sellar radiography showed a disappearance of the posterior clinoid while magnetic resonance imaging revealed the existence of a bilocular, circular, homogeneous lesion of the sphenoid sinus 3 cm in diameter with a posterior and lateral extension. The diagnosis of mucocele was confirmed by surgical treatment, allowing drainage of the mucocele through a transsphenoidal approach. The drained material was composed of sinus epithelium containing many polynuclear and resorptive cells. Postoperatively, the symptoms decreased dramatically, leading to full recovery of visual function and disappearance of the headaches. Apart from the tumor recurrence, the mucocele of the sphenoid sinus can be evoked as a possible long term complication of transsphenoidal surgery for pituitary adenoma. PMID- 10682933 TI - Spontaneous epidural hematoma following a shunt in an infant with congenital factor X deficiency. Case report and literature review. AB - The authors describe a case of an infant with congenital factor X deficiency. The patient presented with a central nervous system hemorrhage followed by hydrocephalus. He underwent a ventriculoperitoneal shunt and, during the postoperative period, developed a spontaneous epidural hematoma, which was evacuated. The clinical and pathophysiological aspects of this case are discussed based on a literature review. PMID- 10682934 TI - A sensitive brain retractor for neurosurgery. PMID- 10682935 TI - Protective effect of metallothionein to ras DNA damage induced by hydrogen peroxide and ferric ion-nitrilotriacetic acid. AB - Metallothionein (MT) is a strong antioxidant, due to a large number of thiol groups in the MT molecule and MT has been found in the nucleus. To investigate whether MT can directly protect DNA from damage induced by hydroxyl radical, the effects of MTs on DNA strand scission due to incubation with ferric ion nitrilotriacetic acid and H2O2 (Fe3+ -NTA/H2O2) were studied. The Fe3+-NTA/H2O2 resulted in a higher rate of deoxyribose degradation, compared to incubation of Fe3+/H2O2, presumably mediated by the formation of hydroxyl radicals (*OH). This degradation was inhibited by either Zn-MT or Cd-MT, but not by Zn2+ or Cd2+ at similar concentrations. The Fe3+ -NTA/H2O2 resulted in a concentration dependent of increase in DNA strand scission. Damage to the sugar-phosphodiester chain was predominant over chemical modifications of the base moieties. Incubation with either Zn-MT or Cd-MT inhibited DNA damage by approximately 50%. Preincubation of MT with EDTA and N-ethylmaleimide, to alkylate sulfhydryl groups of MT, resulted in MT that was no longer able to inhibit DNA damage. These results indicates that MT can protect DNA from hydroxyl radical attack and that the cysteine thiol groups of MT may be involved in its nuclear antioxidant properties. PMID- 10682936 TI - Toxicity and metabolism of malachite green and leucomalachite green during short term feeding to Fischer 344 rats and B6C3F1 mice. AB - Malachite green, an N-methylated diaminotriphenylmethane dye, has been widely used as an antifungal agent in commercial fish hatcheries. Malachite green is reduced to and persists as leucomalachite green in the tissues of fish. Female and male B6C3F1 mice and Fischer 344 rats were fed up to 1200 ppm malachite green or 1160 ppm leucomalachite green for 28 days to determine the toxicity and metabolism of the dyes. Apoptosis in the transitional epithelium of the urinary bladder occurred in all mice fed the highest dose of leucomalachite green. This was not observed with malachite green. Hepatocyte vacuolization was present in rats administered malachite green or leucomalachite green. Rats given leucomalachite green also had apoptotic thyroid follicular epithelial cells. Decreased T4 and increased TSH levels were observed in male rats given leucomalachite green. A comparison of adverse effects suggests that exposure of rats or mice to leucomalachite green causes a greater number of and more severe changes than exposure to malachite green. N-Demethylated and N-oxidized malachite green and leucomalachite green metabolites, including primary arylamines, were detected by high performance liquid chromatography/mass spectrometry in the livers of treated rats. 32P-Postlabeling analyses indicated a single adduct or co eluting adducts in the liver DNA. These data suggest that malachite green and leucomalachite green are metabolized to primary and secondary arylamines in the tissues of rodents and that these derivatives, following subsequent activation, may be responsible for the adverse effects associated with exposure to malachite green. PMID- 10682937 TI - Specific interactions of monotetrahydrofuranic annonaceous acetogenins as inhibitors of mitochondrial complex I. AB - Annonaceous acetogenins (ACG) are a wide group of cytotoxic compounds isolated from plants of the Annonaceae family. Some of them are promising candidates to be a future new generation of antitumor drugs due to the ability to inhibit the NADH:ubiquinone oxidoreductase of the respiratory chain (mitochondrial complex I), main gate of the energy production in the cell. ACG are currently being tested on standard antitumor trials although little is known about the structure activity relationship at the molecular level. On recent studies, the relevance of several parts of the molecule for the inhibitory potency has been evaluated. Due to the great diversity of skeletons included in this family of natural products, previous studies on the presence and distribution of oxygenated groups along the alkyl chain only covered the compounds with different bis-tetrahydrofuranic (bis THF) relative configurations. Therefore, we have investigated the inhibitory action of all the mono-tetrahydrofuranic (mono-THF) acetogenins available, which differ in the oxygenated arrangements along the molecule. Our results show that the hydroxyl and carbonyl groups, placed in the aliphatic chain that links the initial gamma-lactone moiety with the dihydroxylated tetrahydrofuranic ring system, significantly contribute for modulating the inhibitory potency of the ACG through specific effects. PMID- 10682938 TI - Identification of the zinc-binding protein specifically present in male rat liver as carbonic anhydrase III. AB - A zinc (Zn)-binding protein that is present specifically in the livers of male adult rats was detected by HPLC with in-line detection by mass spectrometry (ICP MS). The Zn-binding protein was purified on Sephadex G-75 and G3000SW HPLC columns. and was identified as carbonic anhydrase III (CAIII) based on the amino acid sequence of a peptide obtained on lysyl endopeptidase digestion. CAIII is expressed as one of the major Zn-binding proteins in the livers of male rats in an age-dependent manner, a comparable amount of Zn to that of copper, Zn superoxide dismutase (Cu,Zn-SOD) being bound to CAIII at 8 weeks of age. Castration at 4 or 8 weeks of age was shown to reduce Zn bound to CAIII to 47.5% of the sham-operated control level, suggesting that the sex-dependent expression of CAIII is partly regulated by a sex hormone, androgen. The concentration of CAIII in the livers of Long-Evans rats with a cinnamon-like coat color (LEC rats), an animal model of Wilson disease, was also estimated as Zn bound to CAIII and shown to be lower than that in Wistar rats before the onset of hepatitis. The concentration of CAIII was decreased specifically by repeated injections of cupric ions without the Cu,Zn-SOD concentration being affected. PMID- 10682939 TI - Daytime sleepiness predicts mortality and cardiovascular disease in older adults. The Cardiovascular Health Study Research Group. AB - INTRODUCTION: As part of the baseline examination in the Cardiovascular Health Study, sleep disturbance symptoms including snoring and daytime sleepiness, were assessed as potential risk factors or precipitants of cardiovascular disease (CVD). Because of the association of sleep disturbance with poorer health and the possible associations of sleep apnea with CVD, we hypothesized that those with poorer sleep or daytime sleepiness may be at increased risk of mortality or incident CVD. SETTING: Participants (n = 5888) were recruited in 1989, with an additional minority cohort recruited in 1993, in four US communities for a cohort study designed to evaluate risk factors for cardiovascular disease. METHODS: An interview-administered questionnaire regarding health and sleep habits with ongoing ascertainment of total mortality and cardiovascular disease morbidity and mortality, including total CVD morbidity and mortality, incident myocardial infarction, and congestive heart failure. RESULTS: Daytime sleepiness was the only sleep symptom that was significantly associated with mortality in both men and women. The unadjusted hazard ratio was 2.12 (1.66, 2.72) in women and 1.40 (1.12, 1.73) in men. Men who reported difficulty falling asleep also had an increased mortality rate (HR = 1.43 (1.14, 1.80)) which was not seen in women. The risks were attenuated with adjustment for age but remained significant for daytime sleepiness in women (HR = 1.82 (1.42, 2.34)) and for difficulty falling asleep in men. (HR = 1.29 (1.03, 1.63)). Frequent awakenings, early morning awakening, and snoring were not associated with a significantly increased risk of mortality in these older men and women. Crude event rates were evaluated for total incident cardiovascular morbidity and mortality, incident myocardial infarction, and incident congestive heart failure (CHF). Incident CVD rates were higher in both men and women with daytime sleepiness. The aged adjusted HR was 1.35 (95% CI = 1.03, 1.76) in men and was 1.66 (95% CI = 1.28, 2.16) in women. Incident CVD was not higher in those with any other sleep disturbance including snoring. The risk of CVD events associated with daytime sleepiness was attenuated but remained significant in women after adjustment for age. Incident myocardial infarction (MI) rates were also higher in women with daytime sleepiness but were not significantly higher in men. Incident CHF rates were increased in both men and women with daytime sleepiness. In men, the age adjusted HR was 1.49 (95% CI, 1.12- 1.98) and in women, was 2.21 (95% CI, 1.64-2.98). Women reporting both daytime sleepiness and frequent awakening had a hazard ratio of 2.34 (95% CI, 1.66-3.29) for incident CHF compared with those with daytime sleepiness but without frequent awakening. This interaction was not found in men. CONCLUSIONS: In this study, daytime sleepiness was the only sleep disturbance symptom that was associated with mortality, incident CVD morbidity and mortality, MI, and CHF. These findings were stronger in women than men, i.e., the associations persisted for mortality, CVD, and CHF in women after adjustment for age and other factors. Thus, a report of daytime sleepiness identifies older adults at increased risk for total and cardiovascular mortality, and is an independent risk factor in women. PMID- 10682940 TI - Delivery of preventive services to older black patients using neighborhood health centers. AB - OBJECTIVES: Older black patients are at risk for underutilization of preventive services. Our objectives were to assess the delivery of five preventive services in Title 330-funded health centers in low income neighborhoods in Cleveland, Ohio, and to determine the association of health system factors and health status with the delivery of these services. DESIGN: A cross-sectional study. SETTING: Four neighborhood health centers in low income neighborhoods of Cleveland, Ohio. PARTICIPANTS: A total of 683 black men and women, aged 70 and older, who regarded the health center as their primary source of outpatient care. MEASUREMENTS: Demographic characteristics, independence in basic and instrumental activities of daily living, comorbidity scores, and perceived access were determined by telephone interview. We reviewed charts to determine whether each of five preventive service goals were obtained: influenza vaccination within 1 year; pneumococcal vaccination at any time; mammography within 2 years; Papanicolau screening within 1 year or twice at any time in the past with documentation of normal results; and fecal occult blood testing within 2 years. RESULTS: The defined goals for influenza vaccination, pneumococcal vaccination, mammography, Papanicolau screening, and fecal occult blood testing were achieved for 59%, 64%, 59%, 51%, and 17% of patients, respectively. Influenza and pneumococcal vaccines were obtained more often in persons with greater comorbidity. Mammography and Papanicolau smear were obtained more often in patients without of ADL or IADL impairments. The four clinical sites varied substantially in the delivery of each preventive service. More frequent office visits were associated with greater delivery of all five preventive services. This relationship persisted in multivariable analyses controlling for health status and clinical site. CONCLUSIONS: This study shows that Title 330 federally supported neighborhood health center sites providing primary care to older blacks in Cleveland achieved high rates of performance in four of the five recommended preventive services. In addition, preventive services practices were associated with prognostically relevant health status information. The frequency of office visits was related strongly and consistently to the performance of the various preventive services, indicating that more, not fewer, office visits may be necessary to achieve Healthy People 2000 targets. J Am Geriatr Soc 48:124-130, 2000. Key words: preventive services; blacks; access to care; geriatrics; primary care PMID- 10682941 TI - Long-term exercise and its effect on balance in older, osteoarthritic adults: results from the Fitness, Arthritis, and Seniors Trial (FAST). AB - OBJECTIVES: To examine the effects of 18-month aerobic walking and strength training programs on static postural stability among older adults with knee osteoarthritis. DESIGN: Randomized, single-blind, clinical trial of therapeutic exercise. SETTING: Both center-based (university) and home-based. PARTICIPANTS: A cohort of 103 older adults (age = 60 years) with knee osteoarthritis who were participants in a large (n = 439) clinical trial and who were randomly assigned to undergo biomechanical testing. INTERVENTION: An 18-month center- (3 months) and home-based (15 months) therapeutic exercise program. The subjects were randomized to one of three treatment arms: (1) aerobic walking; (2) health education control; or (3) weight training. MEASUREMENTS: Force platform static balance measures of average length (Rm) of the center of pressure (COP), average velocity (Vel) of the COP, elliptical area (Ae) of the COP, and balance time (T). Measures were made under four conditions: eyes open, double- and single-leg stances and eyes closed, double- and single-leg stances. RESULTS: In the eyes closed, double-leg stance condition, both the aerobic and weight training groups demonstrated significantly better sway measures relative to the health education group. The aerobic group also demonstrated better balance in the eyes open, single-leg stance condition. CONCLUSIONS: Our results suggest that long-term weight training and aerobic walking programs significantly improve postural sway in older, osteoarthritic adults, thereby decreasing the likelihood of larger postural sway disturbances relative to a control group. PMID- 10682942 TI - The relationship between aerobic exercise capacity and circulating IGF-1 levels in healthy men and women. AB - OBJECTIVES: To determine whether aerobic capacity is associated independently with insulin-like growth factor-I (IGF-1) levels in healthy community-dwelling men and women. SETTING: The Baltimore Longitudinal Study on Aging (BLSA). DESIGN: A cross-sectional analysis of data from the population-based cohort of the Baltimore Longitudinal Study of Aging (BLSA). PARTICIPANTS: We studied 181 men and 92 women aged 20 to 93 years, volunteers in the Baltimore Longitudinal Study on Aging (BLSA). Subjects were free of endocrine, renal, hepatic, gastrointestinal, or cardiac diseases, and they were taking no medications known to interfere with the growth hormone-IGF-1 axis. MEASUREMENTS: All subjects underwent a single measurement of serum IGF-1 in the fasting state, as well as peak VO2 determinations during maximal treadmill exercise testing performed within one visit of the IGF-1 determination. Dual energy X-ray absorptiometry (DEXA) scans were performed in a subset of 171 subjects (64 women and 107 men) for determination of fat free mass (FFM). RESULTS: In the pooled group of women and men, univariate regression analysis revealed that age was correlated strongly with decreasing IGF-1 levels (r = -0.53, P < .001) and with peak VO2r = -0.56, P < .001). IGF-1 levels were also significantly correlated with peak VO2 (r = 0.29, P < .001). There were no significant gender-related differences in these relationships. On multivariate analysis, age (beta = -0.54, P < .001), but not peak VO2 (P = -0.01, P = .840), remained strongly associated with IGF-1 levels. After adjustment of peak VO2 for FFM in subjects with DEXA scans, results were similar. CONCLUSIONS: These findings indicate that although both peak aerobic capacity and circulating IGF-1 levels decline with age, aerobic capacity is not independently related to circulating IGF-1 in healthy men and women across the adult life span. PMID- 10682943 TI - Determinants of exercise tolerance after acute myocardial infarction in older persons. AB - OBJECTIVES: Exercise tolerance is reduced with advancing age. Identification of potentially reversible determinants of the age-related decrement in exercise tolerance, which remain largely unexplored in older subjects and in patients recovering from a recent myocardial infarction (MI), may have useful therapeutic implications. The objective of this study was to identify the independent determinants of exercise tolerance in older patients with a recent MI. DESIGN, SETTING, AND PARTICIPANTS: Data is from baseline assessment of 265 post-MI patients (age range 45-85 years) enrolled in the Cardiac Rehabilitation in Advanced Age randomized, controlled trial. Patients with major comorbidities or severe MI complications were excluded from the trial. Exercise tolerance was determined from symptom-limited exercise testing and expressed as total work capacity (TWC, kg.m) or peak oxygen consumption (VO2peak, mL/kg/min). The associations between both TWC and VO2peak and baseline demographic, social, clinical, and neuropsychological variables and an index of health-related quality of life were determined with univariate and multivariate analysis. RESULTS: With univariate analysis, TWC decreased by 1285 kg.m per decade of increasing age between 45 and 85 years of age. With multivariate analysis, TWC decreased by 922 kg.m per decade. Increasing age (P < .001), female gender (P < .001), a small body surface area (P < .001), a low level of usual physical exercise before MI (P < .002), and the presence of post-MI depressive symptoms (P < .024) were independently associated with a lower TWC. The same factors, in addition to a small arm muscle area (P < .002), were also independently associated with a lower VO2peak. CONCLUSIONS: Age per se accounts for approximately 70% of the age related decay in TWC or VO2peak. However, the inclusion of modifiable factors such as physical exercise and depression in the prediction model reinforces the importance of a multidimensional approach to the evaluation and treatment of older patients with a recent MI. PMID- 10682944 TI - Appropriateness of the decision to transfer nursing facility residents to the hospital. AB - OBJECTIVES: To develop and test a standardized instrument, the purpose of which is to assess (1) whether skilled nursing facilities (SNFs) transfer residents to emergency departments (ED) inappropriately, (2) whether residents are admitted to hospitals inappropriately, (3) and factors associated with inappropriate transfers. DESIGN: A structured implicit review (SIR) of medical records. SETTING AND PARTICIPANTS: Using nested random sampling in eight community SNFs, we identified SNF and hospital records of 100 unscheduled transfers to one of 10 hospitals. MEASUREMENTS: Seven trained physician reviewers assessed appropriateness using a SIR form designed for this study (2 independent reviews per record, 200 total reviews). We measured interrater reliability with kappa statistics and used bivariate analysis to identify factors associated with assessment that transfer was inappropriate. RESULTS: In 36% of ED transfers and 40% of hospital admissions, both reviewers agreed that transfer/admit was inappropriate, meaning the resident could have been cared for safely at a lower level of care. Agreement was high for both ED (percent agreement 84%, kappa .678) and hospital (percent agreement 89%, kappa .779). When advance directives were considered, both reviewers rated 44% of ED transfers and 45% of admissions inappropriate. Factors associated with inappropriateness included the perceptions that: (1) poor quality of care contributed to transfer need, (2) needed services would typically be available in outpatient settings, and (3) the chief complaint did not warrant hospitalization. CONCLUSIONS: Inappropriate transfers are a potentially large problem. Some inappropriate transfers may be associated with poor quality of care in SNFs. This study demonstrates that structured implicit review meets criteria for reliable assessment of inappropriate transfer rates. Structured implicit review may be a valuable tool for identifying inappropriate transfers from SNFs to EDs and hospitals. PMID- 10682945 TI - Functional status before hospitalization in acutely ill older adults: validity and clinical importance of retrospective reports. AB - OBJECTIVES: Retrospective reports of patients' functional status before hospital admission are often used in longitudinal studies and by clinicians caring for hospitalized patients. However, the validity of these reports has not been established. Our aim was to examine the validity of retrospective reports by testing hypotheses about the relationships these measures would have with other clinical measures if they were valid. DESIGN: A prospective cohort study. PARTICIPANTS AND SETTING: A total of 2877 older patients (mean age 81, 36% women) hospitalized on the general medical service at two hospitals. For 1953 of the subjects, the patient was the primary respondent, whereas for 924 subjects, a surrogate was the primary respondent. MEASUREMENTS: Shortly after hospital admission, patients or surrogates reported whether the patient was independent in each of five activities of daily living (ADLs) on admission and at baseline 2 weeks before admission. Outcome measures included reported independence in each ADL 3 months after the hospitalization and survival to 1 year. RESULTS: Patients' retrospective reports of their ADL function 2 weeks before admission had a clinically plausible relationship with ADL function at the time of admission, in that patients independent in an ADL on admission rarely reported they were dependent in that ADL 2 weeks before admission (range 2-6%). Surrogates were somewhat more likely than patients to report that patients independent on admission were dependent 2 weeks before admission (range 5-14%). Retrospective reports of prehospitalization ADL function demonstrated strong evidence of predictive validity for both patients' and surrogates' reports. For example, among patients dependent in bathing on admission, patients who were reported as independent 2 weeks before admission were much more likely than those reported as dependent 2 weeks before admission to be independent 3 months after hospitalization (68% vs 20%, P < .001 for patient respondents; 30% vs 5%, P < .001 for surrogate respondents). Similarly, among patients dependent in bathing on hospital admission, survival 1 year after hospitalization was much higher in patients who were independent in bathing 2 weeks before admission than patients who were dependent 2 weeks before admission (76% vs 59%, P < .001 for patient respondents; 60% vs 45%, P < .001 for surrogate respondents). Results were similar for each of the other four ADLs. In a logistic regression model controlling for the number of ADLs reported as dependent on admission, the number of ADLs reported as dependent 2 weeks before admission was significantly associated with 1-year mortality among both patient (odds ratio (OR) = 1.39 per dependent ADL, 95% confidence interval (CI) - 1.26-1.54) and surrogate (OR = 1.14, 95% CI = 1.06-1.24) respondents. CONCLUSIONS: Hospitalized patients' assessments of their ability to perform ADLs before their hospitalization have evidence of face and predictive validity. These measures are strong predictors of important health outcomes such as functioning and survival. In particular, among patients dependent in ADL function on hospital admission, these results highlight the prognostic importance of inquiring about the patient's functional status before the onset of the acute illness. PMID- 10682946 TI - Modifiable risk factors predict functional decline among older women: a prospectively validated clinical prediction tool. The Study of Osteoporotic Fractures Research Group. AB - OBJECTIVE: To identify modifiable predictors of functional decline among community-residing older women and to derive and validate a clinical prediction tool for functional decline based only on modifiable predictors. DESIGN: A prospective cohort study. SETTING: Four geographic areas of the United States. PARTICIPANTS: Community-residing women older than age 65 recruited from population-based listings between 1986 and 1988 (n = 6632). MEASUREMENTS: Modifiable predictors were considered to be those that a clinician seeing an older patient for the first time could reasonably expect to change over a 4-year period: benzodiazepine use, depression, low exercise level, low social functioning, body-mass index, poor visual acuity, low bone mineral density, slow gait, and weak grip. Known predictors of functional decline unlikely to be amenable to intervention included age, education, medical comorbidity, cognitive function, smoking history, and presence of previous spine fracture. All variables were measured at baseline; only modifiable predictors were candidates for the prediction tool. Functional decline was defined as loss of ability over the 4 year interval to perform one or more of five vigorous or eight basic daily activities. RESULTS: Slow gait, short-acting benzodiazepine use, depression, low exercise level, and obesity were significant modifiable predictors of functional decline in both vigorous and basic activities. Weak grip predicted functional decline in vigorous activities, whereas long-acting benzodiazepine use and poor visual acuity predicted functional decline in basic activities. A prediction rule based on these eight modifiable predictors classified women in the derivation set into three risk groups for decline in vigorous activities (12%, 25%, and 39% risk) and two risk groups for decline in basic activities (2% and 10% risk). In the validation set, the probabilities of functional decline were nearly identical. CONCLUSIONS: A substantial portion of the variation of functional decline can be attributed to risk factors amenable to intervention over the short term. Using eight modifiable predictors that can be identified in a single office visit, clinicians can identify older women at risk for functional decline. PMID- 10682947 TI - Decision-making capacity to execute a health care proxy: development and testing of guidelines. AB - OBJECTIVE: To evaluate the reliability and validity of guidelines to determine the capacity of nursing home residents to execute a health care proxy (HCP). DESIGN: A cross-sectional study. SETTING: A 750-bed not-for-profit nursing home located in New York City. PARTICIPANTS: A random sample of 200 nursing home residents: average age, 87; 99% white; 83% female; average length of stay, 3.05 years; mean Mini-Mental State Exam (MMSE) score, 15.9. MEASUREMENTS: Demographic characteristics (Minimum Data Set (MDS)); function and cognitive status (Institutional Comprehensive Assessment and Referral Evaluation (INCARE)); Reisberg Dementia Staging; MMSE; Minimum Data Set-Cognitive Performance Scale (MDS-COGS)); an investigator-developed measure of a nursing home resident's capacity to execute a health care proxy (Health Care Proxy (HCP) Guidelines.) RESULTS: The internal consistency of the decision-making scales in the HCP Guidelines, paraphrased recall and recognition, reached acceptable levels, alphas of .85 and .73, respectively. Interrater reliability estimates were .92 and .94, respectively, for the recall and recognition scales; test-retest reliability estimates were .83 and .90. The discriminant validity of these scales is promising. For example, the MMSE correlation was .51 with the Recall scale and .57 with the Recognition scale. Of residents with severe cognitive impairment (MMSE < 10), 71% completed 50% or more of the scaled items in the HCP guidelines and 95% consistently named a proxy. CONCLUSIONS: Seventy-three percent of testable residents, approximately three-quarters of whom were cognitively impaired, evidenced sufficient capacity to execute an HCP. Of residents with severe cognitive impairment, the HCP guidelines are potentially useful in identifying those with the capacity to execute a HCP. The guidelines are more predictive than the MMSE in identifying residents able to execute a HCP. PMID- 10682948 TI - Physicians are less willing to treat suicidal ideation in older patients. AB - OBJECTIVE: Older adults have the highest rate of suicide of any age group, and reducing the number of late-life suicides has become a national priority. The objective of this study was to determine if an age bias exists among primary care physicians when they contemplate treating suicidal patients. DESIGN: Primary care providers were mailed one of two case vignettes of a suicidal, depressed patient. The only difference between the two vignettes was the age of the patient (38 or 78 years old) and employment status (employed vs retired as a factory worker). A questionnaire was included to determine provider recognition of suicidal ideation, and a scale was designed to detect willingness to treat the vignette patient. SETTING/PARTICIPANTS: Physicians were selected randomly from the University of California, San Francisco physician roster and invited to participate in the study. A total of 342 physicians (63% response rate), including specialists, responded to the mailings. For this study, the responses of 215 primary care physicians were analyzed. INTERVENTION: The randomly assigned experimental group received a vignette of a geriatric, retired patient who was depressed and suicidal (n = 100 participants). The control group received an identical but younger, employed patient (n = 115 participants). MEASUREMENTS: A 21-item Suicidal Patient Treatment Scale measured willingness to treat the suicidal patient. RESULTS: The physicians in this study recognized depression and suicidal risk in both the adult and the geriatric vignette, but they reported less willingness to treat the older suicidal patient compared with the younger patient. The physicians were more likely to feel that suicidal ideation on the part of the older patient was rational and normal. They were less willing to use therapeutic strategies to help the older patient, and they were not optimistic that psychiatrists or psychologists could help the suicidal patient. CONCLUSIONS: This study suggests that primary care physicians are capable of recognizing suicidal ideation but are less willing to treat it if the patient is older and retired. Future research needs to determine etiologic factors for this age bias. PMID- 10682949 TI - Aging is associated with endothelial dysfunction in the human forearm vasculature. AB - OBJECTIVE: Our objective was to examine the role of the endothelium in maintaining vascular tone in the basal as well as in the contracted state during aging. DESIGN/PARTICIPANTS: Responses to brachial artery infusion of acetylcholine in presence and absence of NG-nitro-L-arginine methyl ester (L NAME) and to angiotensin II were studied in 11 young and 12 old white subjects. MEASUREMENTS: Strain gauge plethysmography was used to measure forearm vascular resistance (FVR). The dose of acetylcholine at 50% maximal observed decrease in forearm vascular resistance (EC50) was significantly higher (11.0 +/- 1.59 vs 7.07 +/- .65 microg/min, respectively; mean +/-SEM; P < .05) and the FVR at maximal acetylcholine effect (Emax) remained greater (12.6 +/- 1.75 vs 7.15 +/- 1.25 mm Hg/100 mL tissue volume/min; P < .02) in old compared with young subjects. Acetylcholine effect was significantly reversed by concomitant administration of L-NAME, as indicated by the increase in EC50 (old, 20.2 +/- 3.69; young, 11.9 +/- 1.68 microg/min). RESULTS: There was no age-related difference in sodium nitroprusside-induced decrease in FVR. The EC50 and Emax for angiotensin II-mediated increase in FVR were 7.87 +/- 1.15 and 8.36 +/- 1.00 ng/min (EC50) and 5.30 +/- .67 vs 6.56 +/-1.25 mm Hg/100 mL tissue volume/min (Emax), and these were not different in old and young subjects, respectively. CONCLUSIONS: These data indicate that aging is associated with impaired endothelial- dependent vascular relaxation and that this is selective, with no age-related change in endothelial-independent vascular relaxation or angiotensin II-mediated vascular contraction. PMID- 10682950 TI - The effects of childhood residence in Japan and testing language on cognitive performance in late life among Japanese American men in Hawaii. AB - OBJECTIVES: To examine the association of years spent in Japan during childhood with cognitive test performance in late life among Japanese American men, and to assess the influence of the language used for testing on this association. DESIGN: A cross-sectional study. SETTING/PARTICIPANTS: A total of 3734 Japanese American men, aged 71-93 years, who were first- or second-generation migrants and living on Oahu Island, Hawaii. MEASUREMENTS: The outcome variable was cognitive test performance assessed using the Cognitive Abilities Screening Instrument (CASI), which was developed for cross-cultural studies of cognitive impairment. The explanatory variable of main interest was the number of years spent in Japan during school-age childhood years (ages 6-17). The associations of CASI scores with childhood years in Japan was evaluated using a stepwise multiple linear regression model in which a total of 40 potential confounders were included as covariates. RESULTS: In the total sample, there was an inverse association between CASI scores and middle childhood years in Japan. This association remained significant after controlling for age, education, socioeconomic status, traditional Japanese food consumption, pulmonary function, apolipoprotein E4, proficiency in speaking Japanese, and other possible confounders. When data were analyzed separately for subgroups according to the language preferred at testing (English or Japanese), associations between childhood years in Japan and CASI scores were in opposite directions negative for the group tested in English and positive for the group tested in Japanese. The interaction between the testing language and childhood years in Japan was statistically significant. CONCLUSIONS: There was an inverse association between years spent in Japan during school-age years of childhood and cognitive test performance in late life. This association could not be accounted for by age, education, or other confounding factors. However, this finding was not observed in participants who preferred being tested in Japanese. To assess cognitive test performance in older people, it is of prime importance to use the most optimal language for testing, usually the subject's native language. PMID- 10682951 TI - The high prevalence of depression and dementia in elder abuse or neglect. AB - BACKGROUND: The risk factors for mistreatment of older people include age, race, low income, functional or cognitive impairment, a history of violence, and recent stressful events. There is little information in the literature concerning the clinical profile of mistreated older people. OBJECTIVES: To describe the characteristics of abused or neglected patients and to compare the prevalence of depression and dementia in neglected patients with that of patients referred for other reasons. DESIGN: A case control study. SETTING: Baylor College of Medicine Geriatrics Clinic at the Harris County Hospital District (Houston, Texas). PATIENTS: Forty-seven older persons referred for neglect and 97 referred for other reasons. INTERVENTION: Comprehensive geriatric assessment. MEASUREMENTS: Standard geriatric assessment tools. RESULTS: There was a statistically significant higher prevalence of depression (62% vs 12%) and dementia (51% vs 30%) in victims of self-neglect compared to patients referred for other reasons. CONCLUSIONS: This is the first primary data study that highlights a high prevalence of depression as well as dementia in mistreated older people. Geriatric clinicians should rule out elder neglect or abuse in their depressed or demented patients. PMID- 10682952 TI - Nutritional intake monitoring for nursing home residents: a comparison of staff documentation, direct observation, and photography methods. AB - BACKGROUND: The current approach to assessing nutritional intake requires nursing home (NH) staff to document total percentage of food and fluid consumed at each meal. Because NH staff tend to significantly overestimate total food intake, methods need to be developed to improve the accuracy of food intake measurement. OBJECTIVE: To compare three methods of assessing the nutritional intake of NH residents. RESEARCH DESIGN: Validation Study. SUBJECTS: Fifty-six NH residents in one facility. MEASURES: Total percentage of food and fluid intake of each resident for each of nine meals, or all three meals for 3 consecutive days, was assessed by: (1) Nursing home staff chart documentation, (2) Research staff documentation according to direct observations, and (3) Research staff documentation according to photographs of residents' trays before and after each meal. RESULTS: Research staff documentation of total intake and intake of all individual food and fluid items was similar for the direct observation and photography methods. In comparison with these two methods, NH staff documentation reflected a significant overestimate (22%) of residents' total intake levels. In addition, NH staff failed to identify the more than half (53%) of those residents whose intake levels were equal to or below 75% for most meals. CONCLUSIONS: The photography method of nutritional assessment yielded the same information as direct observations by research staff, and both of these methods showed the intake levels of NH residents to be significantly lower than the intake levels documented by NH staff. The photography method also has several advantages over a documentation system that relies on an observer to be present to record food and fluid intake levels. PMID- 10682953 TI - Health care for older persons, a country profile: The Netherlands. PMID- 10682954 TI - The implementation of the EverCare demonstration project. AB - EverCare represents a creative approach to providing medical services to long stay nursing home patients. It offers a capitated package of Medicare-covered services with more intensive primary care provided by nurse practitioners. The program's underlying premise is that better primary care will result in reduced hospital use. This work examines the implementation of the program in six locations. It identifies some of the issues that must be addressed if the program is to succeed both operationally and financially. PMID- 10682955 TI - The use of oral anticoagulants (warfarin) in older people. AGS Clinical Practices Committee. American Geriatric Society. PMID- 10682956 TI - Ask about daytime sleepiness! PMID- 10682957 TI - Inappropriate hospitalization of nursing facility residents: a symptom of a sick system of care for frail older people. PMID- 10682958 TI - Hydroxyurea-induced cutaneous ulceration in older patients. PMID- 10682959 TI - Subclinical hypothyroidism in a biethnic, urban community. PMID- 10682960 TI - Subclinical hypothyroidism. PMID- 10682962 TI - Subclinical hypothyroidism in a bi-ethnic, urban community PMID- 10682961 TI - Subclinical hypothyroidism in a bi-ethnic, urban community. PMID- 10682963 TI - Optical coherence tomography and scanning laser polarimetry in normal, ocular hypertensive, and glaucomatous eyes. AB - PURPOSE: To evaluate the relationship between visual function and retinal nerve fiber layer measurements obtained with scanning laser polarimetry and optical coherence tomography in a masked, prospective trial. METHODS: Consecutive normal, ocular hypertensive, and glaucomatous subjects who met inclusion and exclusion criteria were evaluated. Complete ophthalmologic examination, disk photography, scanning laser polarimetry, optical coherence tomography, and automated achromatic perimetry were performed. RESULTS: Seventy-eight eyes of 78 patients (17 normal, 23 ocular hypertensive, and 38 glaucomatous) were enrolled (mean age, 56.8+/-11.5 years; range, 26 to 75 years). Eyes with glaucoma had significantly greater neural network scores on scanning laser polarimetry and lower maximum modulation, ellipse modulation, and mean retinal nerve fiber layer thickness measured with optical coherence tomography compared with normal and ocular hypertensive eyes, respectively (all P<.005). Significant associations were observed between neural network number (r = -.51, r = .03), maximum modulation (r = .39, r = -.32), ellipse modulation (r = .36, r = -.28), and optical coherence tomography-generated retinal nerve fiber layer thickness (r = .68, r = -.59) and visual field mean deviation and corrected pattern standard deviation, respectively. All scanning laser polarimetry parameters were significantly associated with optical coherence tomography-generated retinal nerve fiber layer thickness. CONCLUSION: Optical coherence tomography and scanning laser polarimetry were capable of differentiating glaucomatous from nonglaucomatous populations in this cohort; however considerable measurement overlap was observed among normal, ocular hypertensive, and glaucomatous eyes. Retinal nerve fiber layer structural measurements demonstrated good correlation with visual function, and retinal nerve fiber layer thickness by optical coherence tomography correlated with retardation measurements by scanning laser polarimetry. PMID- 10682964 TI - The long-term safety and efficacy of brinzolamide 1.0% (azopt) in patients with primary open-angle glaucoma or ocular hypertension. The Brinzolamide Long-Term Therapy Study Group. AB - PURPOSE: Oral carbonic anhydrase inhibitors used to treat glaucoma have significant systemic side effects. Brinzolamide 1.0%, a new topical ocular carbonic anhydrase inhibitor, is effective apparently without significant systemic side effects. This study was performed to establish the long-term safety and efficacy of brinzolamide 1.0% two and three times daily for primary open angle glaucoma and ocular hypertension. METHODS: An 18-month, multicenter, double masked, parallel, controlled study was conducted. Patients were randomized to brinzolamide two or three times daily or timolol 0.5% twice daily in a 2:2:1 ratio (n = 150, 153, and 75, respectively). Intraocular pressure was measured at 8:00 AM at eligibility and months 1, 3, 6, 9, 12, 15, and 18. Efficacy was based on intraocular pressure reduction from baseline. Safety was also evaluated. RESULTS: All regimens produced clinically relevant and statistically significant (P<.05) intraocular pressure reductions from baseline. Mean changes in intraocular pressure trough measurements ranged from -2.7 to -3.9 mm Hg with brinzolamide twice-daily dosing and -2.8 to -3.8 mm Hg three times daily dosing compared with -4.7 to -5.6 mm Hg with timolol. The intraocular pressure reductions with brinzolamide two and three times daily were clinically and statistically equivalent. One hundred forty-four patients were discontinued from the study after randomization with the most common reasons being the occurrence of an adverse event (46), inadequate intraocular pressure control (23), patient decision unrelated to study medication (11), lost to follow-up (16), and noncompliance (9). Adverse events were nonserious and resolved without sequelae. There were no clinically relevant changes in safety parameters. Brinzolamide produced less ocular discomfort (burning/stinging) than timolol, and total carbonic anhydrase inhibition levels remained below that known to cause systemic side effects. CONCLUSION: Brinzolamide produced significant and equivalent reductions in intraocular pressure when dosed two and three times daily for 18 months. Brinzolamide was safe and well tolerated by patients, with minimal ocular discomfort. PMID- 10682965 TI - Dorzolamide and corneal recovery from edema in patients with glaucoma or ocular hypertension. AB - PURPOSE: To investigate whether dorzolamide alters corneal hydration control in patients with glaucoma or ocular hypertension. METHODS: Pachymetry, tonometry, and endothelial cell density were measured by a masked observer in 19 subjects with bilateral glaucoma or ocular hypertension. They were treated with 2% dorzolamide in one eye, and with saline in the other, before wearing contact lenses under patched eyes. Corneal thickness, measured each 30 minutes up to 4.5 hours after contact lens removal, enabled estimation of percentage recovery per hour and time for 95% of corneal thickness recovery for both eyes. Seven patients repeated this test after 1 year of dorzolamide use, and their results were compared with those of the preceding year. RESULTS: After induction of hypoxic corneal edema, there was no significant difference between paired corneas in swelling levels (60.0+/-11.8 and 59.8+/-12.9 microm) (P = .94), time to 95% recovery (440.6+/-255.8 and 445.4+/-186.7 minutes) (P = .93), and percentage recovery per hour (38.1%+/-10.9% and 36.1%+/-9.6%) (P = .40). Subjects followed up after 1 year of dorzolamide use did not differ significantly in values of endothelial cell density, percentage recovery per hour, or time to 95% recovery from those obtained a year before. One subject developed persistent corneal edema after his stress test in the eye treated with dorzolamide. CONCLUSION: There is no significant difference in the recovery from induced corneal edema after either a short-term or 1-year use of dorzolamide in patients with glaucoma or ocular hypertension with a normal corneal endothelium. One patient had persistent corneal edema after the stress test was performed on the dorzolamide-treated eye. PMID- 10682966 TI - Tear and serum soluble leukocyte activation markers in conjunctival allergic diseases. AB - PURPOSE: To measure markers of leukocyte activation in patients with an exclusively ocular inflammatory or bacterial disease. METHODS: Neutrophil myeloperoxidase, eosinophil cationic protein, eosinophil neurotoxin, and soluble interleukin-2 receptor were measured in serum and tears of 17 patients with allergic vernal keratoconjunctivitis, seven with atopic keratoconjunctivitis, 11 with seasonal allergic conjunctivitis, seven with giant papillary conjunctivitis, 13 with rosacea blepharokeratoconjunctivitis, seven with bacterial conjunctivitis, and 13 normal subjects as controls. RESULTS: In serum of patients with vernal and atopic keratoconjunctivitis, levels of eosinophil cationic protein, eosinophil neurotoxin, and interleukin-2 receptor were significantly increased compared with control subjects but were not correlated with the severity of ocular symptoms. In tears of patients with vernal and atopic keratoconjunctivitis and seasonal allergic conjunctivitis, as well as in the nonallergic diseases, rosacea blepharokeratoconjunctivitis and bacterial conjunctivitis, levels of eosinophil cationic protein, neurotoxin, and interleukin-2 receptor were significantly increased compared with control subjects. The highest values of these markers were found in vernal keratoconjunctivitis samples. Neutrophil myeloperoxidase was significantly increased in vernal and atopic keratoconjunctivitis, rosacea blepharokeratoconjunctivitis, and bacterial conjunctivitis. In vernal keratoconjunctivitis, tear markers were correlated to the clinical score of the disease, but not with cytology. CONCLUSIONS: Tear histamine was measured in 10 allergic patients after allergen challenge. Although none of the above markers can be considered specific to a single disease, their measurement may still be useful for the quantification of local cell activation in ocular inflammatory diseases. PMID- 10682967 TI - Genetically distinct autosomal dominant posterior polar cataract in a four generation Japanese family. AB - PURPOSE: To describe the clinical findings of a form of posterior polar cataract in a large Japanese family and to determine whether the posterior polar cataract is causally related to other autosomal dominant cataracts with known genes, chromosomal locations, or both. METHODS: Systemic and ocular histories were obtained and comprehensive ophthalmic examinations were performed in 15 of 37 members of the Japanese family. The posterior polar cataract was transmitted in an autosomal dominant manner through four generations. Although there is some variation in the degree of opacification, the posterior polar cataract in this family is characterized by progressive disk-shaped posterior subcapsular opacities. Genetic linkage analysis was performed with 41 polymorphic microsatellite markers located in chromosomal regions known for linkage to cataracts. Genomic DNA extracted from the 15 individuals was amplified by polymerase chain reaction, the genotype at the marker loci was determined in each family member, and the lod score was calculated at each locus. RESULTS: Significant linkage of the posterior polar cataract was ruled out from the following 10 loci or chromosomal regions: 16q22 and 1p36, to which two forms of autosomal dominant posterior polar cataract have been assigned: 1q21-q25, 2q33 q35, 13cen, 17p13, 17q11-q12, 17q24, 21q22, and 22q, which are the regions responsible for other autosomal dominant congenital cataracts. CONCLUSIONS: This study confirms the genetic heterogeneity of autosomal dominant posterior polar cataracts and demonstrates that the posterior polar cataract in this Japanese family is phenotypically and genetically distinct from previously mapped cataracts. PMID- 10682968 TI - Viral causes of the acute retinal necrosis syndrome. AB - PURPOSE: The primary goal of this study was to determine the viral cause of the acute retinal necrosis syndrome in 28 patients (30 eyes). A secondary goal was to investigate possible associations between viral cause and patient age, and viral cause and central nervous system disease. METHODS: A retrospective case series in which we reviewed the laboratory results and clinical histories of 28 patients (30 eyes) diagnosed with acute retinal necrosis syndrome, from whom vitreous or aqueous specimens were received, for diagnostic evaluation using previously described polymerase chain reaction-based assays. RESULTS: Varicella-zoster virus, herpes simplex virus, and cytomegalovirus (CMV) DNA were detected in aqueous and/or vitreous specimens from 27 of 28 patients (29 of 30 eyes with a clinical history of acute retinal necrosis syndrome). No sample was positive for DNA from more than one virus. Varicella-zoster virus DNA was detected in 13 patients (15 eyes). Median age was 57 years. Herpes simplex virus type 1 DNA was detected in seven patients (seven eyes). Median age was 47 years. Six of these patients had a history of herpes simplex virus encephalitis. Herpes simplex virus type 2 DNA was detected in six patients (six eyes). Median age was 20 years. Three of these patients had a likely history of meningitis. Cytomegalovirus DNA was detected in one patient who was immunosuppressed iatrogenically. No viral DNA was detected in one patient from whom a sample was taken after 6 weeks of acyclovir therapy. CONCLUSIONS: The data suggest that varicella-zoster virus or herpes simplex virus type 1 cause acute retinal necrosis syndrome in patients older than 25 years, whereas herpes simplex virus type 2 causes acute retinal necrosis in patients younger than 25 years. A history of central nervous system infection in a patient with acute retinal necrosis syndrome suggests that herpes simplex virus is likely to be the viral cause. PMID- 10682969 TI - Association of HLA with Vogt-Koyanagi-Harada syndrome in Koreans. AB - PURPOSE: To study the distribution of human leukocyte antigen HLA-A/B antigens and HLA-DR/-DQ/-DP alleles and to investigate the immunogenetic background of Korean patients with Vogt-Koyanagi-Harada (VKH) syndrome and clinical course with different types of HLA. METHODS: Human leukocyte antigen typings were performed in 18 Korean patients with VKH syndrome and in 128 healthy control subjects. HLA A/B loci serologic typing was performed according to the standard microlymphocytotoxicity technique. DNA was extracted through the salting out method, and HLA-DR phenotyping and HLA-DR4, HLA-DQ, and HLA-DP subtyping were performed with the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. RESULTS: Among HLA-A/B antigens typed by the standard microlymphocytotoxicity method, the frequencies of HLA-A31 (RR = 6.1, P<1x10(-2)) and HLA-B55 (RR = 15.8, P<.05) were significantly increased in the patient group compared with the control group. Among HLA-DR/-DQ/-DP alleles subtyped by DNA methods, the frequencies of HLA-DRB1*04 (RR = 45.1, P<1x10(-7)) and HLA-DRB1*07 (RR = 3.2, P<.05) were significantly increased. However, significant decreases in HLA-DRB1*08 (RR = .1, P<.05), HLA-DRB1*13 (RR = .1, P<.05), and HLA-DRB1*14 (RR = .1, P<.05) frequencies were observed. The result of HLA-DR, HLA-DQ, and HLA-DP subtyping showed the significant increase in DRB1*0405 (RR = 45.1, P<1x10(-7)), DQA1*0302 (RR = 12.0, P<1x10(-4)), DQB1*0303 (RR = 5.0, P<1x10(-2)), DQB1*0401 (RR = 18.9, P<1x 10-6), and DPB1*0501 (RR = 3.8, P<.05). However, significant decreases in DQA1*0101 (RR = .1, P< .05), DQA10102 (RR = .1, P<1x10(-2)), DQA1*0103 (RR = .1, P<.05), DQA1*0501 (RR = .1, P<1x10(-2)), DQB1*0301 (RR = .1, P<.05), DQB1*0601 (RR = .1, P<.05), DPB1*0201 (RR = .3, P<.05), and DPB1*0401 (RR = .1, P<.05) frequencies were also observed. In patients with DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype, a reduction in visual acuity and ocular complications was common. CONCLUSIONS: These results suggest that HLA DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype is greatly increased and may play the most important role in the development and the clinical course of VKH syndrome in Korean patients. PMID- 10682970 TI - One-year outcomes of panretinal photocoagulation in proliferative diabetic retinopathy. AB - PURPOSE: To describe the clinical features and complications of diabetic retinopathy, visual acuity, and number of repeat treatments after panretinal photocoagulation for proliferative diabetic retinopathy in a tertiary care center. METHODS: A cohort study was conducted with data collection from medical records of patients undergoing panretinal photocoagulation between 1985 and 1995 at the Scheie Eye Institute; 297 eyes of 186 patients were eligible for study. RESULTS: The presence of neovascularization of the disk at baseline, an earlier onset of diabetes, and a shorter duration of disease before panretinal photocoagulation were the strongest risk factors for needing an additional panretinal photocoagulation treatment. Sixty-two percent of eyes with poor visual acuity (< or =20/200) at baseline still had poor visual acuity at 1 year, and 76% with good visual acuity (> or =20/40) at baseline maintained good visual acuity at 1 year. Poor vision at baseline was the only risk factor for having poor vision at 1 year. Vitreous hemorrhage was present in 44% of eyes at baseline. New vitreous hemorrhage developed in 37% of eyes during the first year after panretinal photocoagulation. A traction retinal detachment was present in 4% of eyes at baseline and newly developed in 6% of eyes during follow-up. A repeat panretinal photocoagulation treatment was performed in 39% of eyes after initial treatment. A vitrectomy was performed in 10% of eyes from baseline through the 1 year follow-up visit. CONCLUSIONS: The data from this study are useful for counseling patients with respect to likely visual outcome, possibility of major complications from proliferative diabetic retinopathy, and the chance of undergoing additional laser treatment after panretinal photocoagulation. PMID- 10682971 TI - Optical coherence tomography of the neurosensory retina in rhegmatogenous retinal detachment. AB - PURPOSE: To clarify the pathologic changes of the detached neurosensory retina in rhegmatogenous retinal detachment. METHODS: Retinal images were prospectively examined by optical coherence tomography in 25 eyes of 25 consecutive patients with rhegmatogenous retinal detachment. We excluded the patients whose retinal detachment did not involve the central fovea or patients with poor fixation during optical coherence tomography (OCT) examination. Optical coherence tomography was scanned through the center of the fovea. The patients ranged in age from 15 to 77 years (mean, 45 years; SD, 20 years). The period from onset of subjective symptoms of retinal detachment to OCT ranged from 2 to 60 days (mean, 16 days; SD, 18 days). Optical coherence tomography findings, best-corrected visual acuity, and the height of the retinal detachment at the central fovea were statistically analyzed using ANCOVA (analysis of covariance) and the Mann-Whitney U test. RESULTS: In 25 eyes of 25 patients, OCT of the detached neurosensory retina at and adjacent to the center of the fovea demonstrated normal retinal structure (10 eyes, 40%), intraretinal separation (7 eyes, 28%), and an undulated separated outer retina (8 eyes, 32%). Three statistically significant factors affected best-corrected visual acuity: intraretinal separation (P = .001), intraretinal separation with undulated outer retina (P = .001), and height of retinal detachment at the central fovea (P<.001). Best-corrected visual acuity was significantly worse in the 15 eyes with intraretinal separation with or without an undulated outer retina than in the 10 eyes with retinal thickening but no intraretinal separation (P = .036). The eight eyes with undulated separated outer retina showed significantly higher retinal detachment at the central fovea than the seven eyes with intraretinal separation but no undulated outer retina (P = .009) and the 10 eyes without intraretinal separation (P = .016). The duration from onset of subjective symptoms to OCT was not related to the occurrence of intraretinal separation of the detached retina. CONCLUSIONS: Intraretinal separation of the detached retina occurred frequently and shortly after retinal detachment in rhegmatogenous retinal detachment and was one of the factors associated with poor vision in rhegmatogenous retinal detachment. Best-corrected visual acuity significantly decreased in the highly detached retina. PMID- 10682972 TI - Remodeling of choroidal venous drainage after vortex vein occlusion following scleral buckling for retinal detachment. AB - PURPOSE: To evaluate the choroidal drainage route after scleral buckling for retinal detachment. METHODS: We performed wide-angle indocyanine green angiography with a scanning laser ophthalmoscope in 22 eyes of 22 patients with rhegmatogenous retinal detachment that had been treated with scleral buckling and cryopexy. The 22 eyes had at least one retinal break located posterior to the equator where vortex veins were present. The period between angiography and retinal detachment surgery was less than 3 months in 10 eyes, 3 to 12 months in five eyes, and more than 1 year in seven eyes. RESULTS: At the site of the silicone exoplant and cryopexy for retinal breaks, one vortex vein was occluded in seven eyes and two in five eyes. Choroidal veins were congested in the quadrant of the occluded vortex vein in two of 12 eyes that had angiography less than 3 months after retinal detachment surgery. In 10 of 12 eyes that had angiography 3 months or more after retinal detachment surgery, new drainage routes developed that connected the sector of the occluded vortex veins to that of the intact vortex veins with venovenous anastomoses. No venous congestion was found in the areas of the occluded vortex veins in the 10 eyes. Venous collaterals formed between the superior and inferior vortex in all 10 eyes and between the temporal and nasal vortex in two eyes. CONCLUSIONS: New venous drainage routes, which were connected to the intact vortex veins, formed in eyes with occluded vortex veins resulting from scleral buckling surgery and compensated for choroidal venous congestion. The choroidal veins have a great deal of plasticity that enables remodeling of the drainage routes, depending on the pressure gradient. PMID- 10682973 TI - Intraoperative echographic localization of iodine-125 episcleral plaque for brachytherapy of choroidal melanoma. AB - PURPOSE: To report intraoperative echographic localization of iodine-125 episcleral plaque for brachytherapy of choroidal melanoma. METHODS: In a retrospective study, 117 eyes with medium-sized choroidal melanoma in 117 patients not participating in the Collaborative Ocular Melanoma Study underwent iodine-125 episcleral plaque radiotherapy with intraoperative echographic verification of plaque placement between January 1992 and December 1998 at the Bascom Palmer Eye Institute. RESULTS: After initial plaque placement using standard localization techniques, intraoperative echography demonstrated satisfactory tumor-plaque apposition in 76% of eyes (89 of 117). In the 28 eyes (28 of 117, 24%) that required repositioning of the plaque, the extent of misplacement was less than 1 mm in 10 eyes, 1.1 to 3.0 mm in six eyes, and greater than 3 mm in eight eyes. Two eyes had tilting of the plaque, and in two additional eyes, although the plaque covered all tumor margins, the centration was considered suboptimal. Repositioning was necessary in 1 eye with an anteriorly located tumor (1 of 13, 7.7%) and in 20 eyes with peripapillary or posterior pole tumors (20 of 67, 26.3%). Anteriorly located tumors required plaque repositioning significantly less frequently than did posteriorly located tumors (P = .041). Misalignment involved one tumor margin in 23 eyes and two margins in five eyes. The most commonly misaligned margins were the lateral (35%) and posterior margins (26%). In no case was an anterior marginal misalignment documented. At a mean follow-up of 37 months, no tumor-related death or metastatic disease was noted. Two of the 117 patients (1.7%) had local tumor recurrence and underwent enucleation. CONCLUSIONS: Intraoperative echography is an effective adjunct for localization and confirmation of tumor-plaque relationship. This technique facilitates the identification and correction of suboptimal plaque placement at the time of surgery, potentially minimizing treatment failures. PMID- 10682974 TI - Location, substructure, and composition of basal laminar drusen compared with drusen associated with aging and age-related macular degeneration. AB - PURPOSE: To determine whether basal laminar drusen differ in their location, ultrastructure, or composition from drusen associated with aging and age-related macular degeneration. METHODS: A paraffin-embedded block from an eye of a patient with basal laminar drusen was obtained. Sections were examined immunohistochemically using a battery of antibodies and lectins directed against drusen-associated proteins and glycoconjugates, respectively. Thin sections were examined by electron microscopy and compared with eyes with age-related macular degeneration. RESULTS: Drusen in the eye with basal laminar drusen are located between the basal lamina of the retinal pigment epithelium and the inner collagenous layer of Bruch membrane, just as they are in age-related macular degeneration. Two distinct ultrastructural phenotypes are observed in the eye with basal laminar drusen; their substructure is indistinguishable from drusen phenotypes in age-related macular degeneration. Both basal laminar drusen and drusen associated with age-related macular degeneration are bound by the lectins Ricinis communis agglutinin and Arachis hypogea agglutinin (after neuraminidase digestion) and by antivitronectin, anti-HLA-DR, anti-serum amyloid P, and anti-C5 antibodies, but not by antibodies directed against basement membrane-associated heparan sulfate proteoglycan, laminin, fibrinogen, or collagen type IV. CONCLUSIONS: These data support the notion that cuticular or basal laminar drusen are similar to, and perhaps indistinguishable from, drusen associated with age related macular degeneration and are not nodular or diffuse thickenings of Bruch membrane, as previously suggested. Thus, we suggest basal laminar drusen is a misnomer. This clinical phenotype should be identified as "early adult onset, grouped drusen" or by the eponym "Gass syndrome." Features of basal laminar drusen, such as uniform drusen size, clustered distribution, and angiographic features, do not appear to be related to differences in drusen location, composition, or substructure. PMID- 10682975 TI - Abnormalities in rod photoreceptors, amacrine cells, and horizontal cells in human retinas with retinitis pigmentosa. AB - PURPOSE: To evaluate changes in the rods and amacrine cells and horizontal cells in human retinas with retinitis pigmentosa. METHODS: Seven retinas from patient donors with retinitis pigmentosa and 14 age- and postmortem-matched normal human retinas were processed for immunocytochemistry and confocal microscopy. The following cell-specific antibodies were used: anti-rhodopsin (rods), anti-gamma aminobutyric acid (amacrine cells), anticalbindin (cones and horizontal cells), anti-glial fibrillary acidic protein (astrocytes and reactive Muller cells), and anti-synaptophysin and anti-SV2 (synaptic vesicles). RESULTS: In retinal regions with significant photoreceptor loss, the rods, gamma-aminobutyric acid-positive amacrine cells, and calbindin-positive horizontal cells had undergone neurite sprouting. The rod, amacrine and horizontal cell neurites were associated with the surfaces of glial fibrillary acidic protein-immunoreactive Muller cells. Most rod neurites that projected into the inner retina contacted the somata of gamma aminobutyric acid-positive amacrine cells. CONCLUSIONS: Rods, amacrine and horizontal cells undergo neurite sprouting in human retinas with retinitis pigmentosa. These changes in the retinal neurons may contribute to the electroretinographic abnormalities and progressive decline in vision noted by patients with retinitis pigmentosa. These alterations may also complicate strategies for treatment of retinitis pigmentosa. PMID- 10682976 TI - Retinal micromovements, the visual line, and Donders' law. AB - PURPOSE: To report the relationship of the retinal micromovements to the visual line and to confirm the validity of Donders' Law. METHODS: Two video cameras suspended from a headband were used to record eye (video-oculography) and head movements. Eye positions in held gaze and following various trajectories to a target were recorded in five normal, young subjects. The videotapes were analyzed off-line using a computer algorithm. RESULTS: Retinal micromovements cause the visual line to trace a zigzag pathway across the foveola, which has an approximate diameter of 350 microm (about 2 degrees). The mean micromovement was about 10 microm in 33.3 msec. The cumulative effect of successive micromovements may move the visual line across the foveola from edge to edge depending on the elapsed time. When the visual line reaches the edge of the foveola it changes its direction. When the eye resets to the same target by different trajectories, the visual line may alight up to about 350 microm from its original location anywhere within the foveola. CONCLUSIONS: Donders' Law is upheld because for each direction of gaze, and regardless of the trajectory used to reach that direction of gaze, the retina has a constant orientation to an index head plane at any given moment in time. Failure to consider that the micromovements cause a shift in the position of the visual line within the foveola may account for the exceptions to Donders' Law found by contemporary researchers using invasive recording techniques. PMID- 10682977 TI - The human leukocyte antigen complex and chronic ocular inflammatory disorders. AB - PURPOSE: To review the role of gene products from the human leukocyte antigen (HLA) complex in the normal functioning of the immune system, ocular inflammation, and models of autoimmunity. METHOD: A review of recently published reports. RESULTS: Many chronic ocular inflammatory diseases are associated with specific alleles of the HLA complex. Understanding how HLA gene products function normally provides clues to the mechanism of disease associations. In the thymus, these molecules control the shape of the developing T-cell repertoire, leading to self-tolerance. In the periphery, HLA molecules bind and present peptide fragments to T cells, leading to a variety of effector functions. Although effector functions are for the most part beneficial, models are reviewed in which peptide-HLA interactions lead to T-cell responses with pathologic consequences. Herpes stromal keratitis is an informative animal model highlighting the role of self-tolerance, infection, and molecular mimicry in the development of autoimmunity. CONCLUSIONS: Human leukocyte antigen gene products may be associated with chronic inflammatory disorders through the unique presentation of "disease-inducing" peptides or the development of a T-cell repertoire prone to autoreactivity and molecular mimicry. PMID- 10682978 TI - Conjunctival incision for primary conjunctivodacryocystorhinostomy with Jones tube. AB - PURPOSE: To study a conjunctival incision approach for primary conjunctivodacryocystorhinostomy with Jones tube insertion. METHODS: This technique was applied throughout an 8-year period on 13 adult patients; each presented with complete lacrimal obstruction at the common canaliculus and excessive tearing. RESULTS: Surgical outcome was successful, with no external facial scarring. Follow-up ranged from 9 months to 7 years (median, 4.5 years). Complications included a tube exchange in one patient and occasional tearing in another. CONCLUSION: A conjunctival incision approach to conjunctivodacryocystorhinostomy (CDCR) with Jones tube insertion is an alternative technique for adult patients, especially those known (or potential) keloid-former cases, or those preferring not to undergo laser treatment and wanting to avoid an external facial scar. PMID- 10682979 TI - Primary localized conjunctival amyloidosis presenting with recurrence of subconjunctival hemorrhage. AB - PURPOSE: To report the ocular presentation and histopathology of a patient with primary localized conjunctival amyloidosis. METHODS: A 38-year-old woman presented with a recurrence of episodes of severe bilateral subconjunctival hemorrhage. Ocular examination revealed yellowish, marked folding and redundancy of the conjunctiva in the inferior cul-de-sac of each eye. RESULTS: After two initial conjunctival biopsies that showed only chronic inflammation, a third biopsy revealed the presence of amyloid in the substantia propria of the conjunctiva. CONCLUSION: Primary localized conjunctival amyloidosis is rare and usually diagnosed histologically instead of clinically. Recurrence of subconjunctival hemorrhage may be the initial presentation. Evaluation for systemic diseases is advised, though the results of the examination are almost always negative. PMID- 10682980 TI - Localized infection by Serratia marcescens simulating a conjunctival neoplasm. AB - PURPOSE: To report a Serratia marcescens infection that clinically simulated a conjunctival neoplasm. METHOD: Case report. RESULTS: A healthy 80-year-old man without contact lenses presented with a pink-yellow conjunctival mass that resembled a solid neoplasm. Stains and cultures of material that exuded from the mass during surgery revealed S. marcescens. Histopathology disclosed an epithelial-lined cyst with macrophages containing S. marcescens. CONCLUSION: Although S. marcescens usually affects the eye as a keratoconjunctivitis in patients with contact lenses, it can also present as a mass simulating a neoplasm in a patient who does not wear contact lenses. PMID- 10682981 TI - A new mutation (A546T) of the betaig-h3 gene responsible for a French lattice corneal dystrophy type IIIA. AB - PURPOSE: To characterize the betaig-h3 gene defect in a French family affected with lattice corneal dystrophy type IIIA (LCDIIIA). METHODS: Histologic examination was performed from corneal buttons of two patients. Genomic DNA was extracted from leukocytes, and exons of the betaig-h3 gene were amplified by polymerase chain reaction to be directly sequenced. RESULTS: Numerous deposits were evident in the stroma and beneath the Bowman membrane, which had all the features of amyloid deposits. Analysis of exon 12 revealed a heterozygous G to A transition on codon 546. CONCLUSION: In contrast to Japanese patients, these French patients affected with LCDIIIA carry a distinct mutation of the betaig-h3 gene (A546T instead of P501T). Therefore, it is unclear whether different mutations could result in the same dystrophy or whether we are dealing with clinical heterogeneity of LCDIIIA. PMID- 10682982 TI - Intracorneal hematoma in Mooren ulceration. AB - PURPOSE: To report a case of intracorneal hematoma occurring in association with Mooren ulceration. METHOD: Case report. RESULTS: In an 81-year-old man with bilateral Mooren ulceration, a dense intracorneal hemorrhage occurred in the right eye secondary to peripheral corneal neovascularization and was followed by slow resolution over a 3-year period. Following subsequent lamellar and penetrating keratoplasty, histopathologic examination demonstrated the association between the stromal neovascularization and the residual interlamellar hemorrhage, as well as phagocytosis of residual hemosiderin by macrophages. CONCLUSION: Intracorneal hematoma with spontaneous resolution has been documented clinically and histopathologically in an eye with Mooren ulceration. PMID- 10682983 TI - Choroidal effusions and hypotony caused by severe anterior lens capsule contraction after cataract surgery. AB - PURPOSE: To report the clinical features and management of two patients with pseudophakic anterior capsule contraction with secondary tractional ciliary body detachments and hypotonous choroidal effusions. METHODS: Case reports. RESULTS: In two eyes of two patients with pseudophakia, severe anterior lens capsule contraction and tractional ciliary body detachments, anterior capsulotomy (one Nd:YAG laser, one surgical), was followed by resolution of the ocular hypotony and resolution/nonrecurrence of the choroidal effusions. In both cases, continuous curvilinear capsulorhexis was used during cataract surgery. CONCLUSION: Anterior capsule contraction following pseudophakia may result in tractional ciliary detachment and secondary ocular hypotony. Radial anterior capsulotomy appeared to be effective in both cases. PMID- 10682984 TI - Syndrome simulating pseudotumor cerebri caused by partial transverse venous sinus obstruction in metastatic prostate cancer. AB - PURPOSE: To report a case of partial transverse venous sinus obstruction causing a syndrome resembling pseudotumor cerebri. METHOD: Case report. A 61-year-old man developed decreased vision, bilateral papilledema, and a highly increased cerebrospinal fluid opening pressure. Brain magnetic resonance imaging (MRI) disclosed a small, extra-axial mass near the torcula, which was dismissed as an incidental meningioma because cerebral angiography showed sinus patency. RESULTS: The patient's vision worsened. Biopsy of the enlarging mass disclosed metastatic prostate cancer. After radiation therapy, the mass shrank, magnetic resonance angiography disclosed reopening of the transverse sinuses, and papilledema resolved, but visual fields remained severely compromised. CONCLUSION: Partial blockage of the dural venous sinus by a small mass near the torcula can cause a sufficient increase in intracranial pressure to produce vision-threatening papilledema. PMID- 10682985 TI - Bilateral blindness as the initial presentation of lymphoma of the sphenoid sinus. AB - PURPOSE: To report a patient with large-cell lymphoma of the sphenoid sinus presenting with bilateral blindness and no other signs or symptoms. METHOD: Case report. A previously healthy 5-year-old boy complained of sudden vision loss without other systemic complaints. RESULTS: Ophthalmologic examination revealed no light perception bilaterally. The pupils of the patient were fixed at 8 mm without reaction to the brightest light stimulus. Systemic examination was unremarkable, and neuroimaging revealed a large sphenoid tumor extending intracranially. Biopsy of the tumor proved to be large-cell lymphoma. CONCLUSION: Large-cell lymphoma affecting children may present initially with blindness, without other systemic symptoms. PMID- 10682986 TI - DNA-based diagnosis of the von Hippel-Lindau syndrome. AB - PURPOSE: To evaluate the etiology of a unilateral hemangioblastoma noted in a male with a family history remarkable only for spine surgery in the proband's father. METHODS: Genomic DNA was isolated from peripheral blood of family members, and the three exons of the von Hippel-Lindau gene were examined for mutations by direct sequencing. RESULTS: A three base pair (bp) deletion in exon 1 of the VHL gene was found in the father and both sons. This in-frame deletion results in the loss of a phenylalanine residue from the von Hippel-Lindau protein product, at amino acid position 76. CONCLUSION: Genetic screening has confirmed that von Hippel-Lindau syndrome is responsible for the hemangioblastoma in the proband. Magnetic resonance imaging scans performed as a consequence of these results indicated spinal tumors present in the father and tumors present in the cerebellum of the proband's sibling. As close, lifelong follow-up is warranted with this disease, this case demonstrates the value of DNA testing in patients with ocular findings consistent with von Hippel-Lindau disease in the absence of a recognized family history. PMID- 10682987 TI - A novel spontaneous missense mutation in VMD2 gene is a cause of a best macular dystrophy sporadic case. AB - PURPOSE: To report the molecular characterization of a novel VMD2 mutation causing a Best macular dystrophy sporadic case. METHODS: All family members underwent ophthalmologic examination and genetic testing by single strand conformation polymorphism analysis and direct sequencing of the VMD2 gene. RESULTS: A single T to G transition at nucleotide 663 was identified in one of the VMD2 gene copies of the patient, which results in a Cys to Trp substitution at position 221 in the corresponding protein (C221W). Sequence analysis of the VMD2 exon 6 of both parents of the patient did not reveal any mutation. CONCLUSION: These data confirm the involvement of the VMD2 gene in Best macular dystrophy onset, even in sporadic cases of the disease, pointing out the relevance of molecular analysis in the diagnosis of this degenerative retinal disease. PMID- 10682988 TI - Serous retinal detachment at the macula in sarcoidosis. AB - PURPOSE: To report an unusual case of serous retinal detachment at the macula in a patient with a history of sarcoidosis. METHOD: Case report. In a 44-year-old Caucasian man, the history, clinical features, fluorescein angiography, laboratory and radiological investigations, treatment, and lung autopsy findings are presented. RESULTS: A serous detachment of the retina in the macula of his left eye was observed. He had pulmonary hypertension secondary to active sarcoid granulomas that caused pulmonary angiitis. CONCLUSION: Serous retinal detachment at the macula may occur in association with pulmonary angiitis secondary to sarcoidosis. PMID- 10682989 TI - Optical cross-sectional observation of resolved diabetic macular edema associated with vitreomacular separation. AB - PURPOSE: To describe the resolution of cystoid macular edema associated with vitreomacular separation in a diabetic patient. METHODS: Case report. A 58-year old man who had cataract surgery 3 years earlier developed diabetic macular edema after panretinal laser photocoagulation. For a detailed fundus examination, we performed neodymium: YAG (Nd:YAG) laser capsulotomy in the left eye as the initial management. RESULTS: Two days after the laser capsulotomy, fundus biomicroscopy and B-mode ultrasonography disclosed a vitreomacular separation in the left eye that was not detectable preoperatively. Optical coherence tomography through the macula disclosed a dramatic decrease in the size of intraretinal cystoid spaces with an improvement of visual acuity. Scanning retinal thickness analysis also confirmed the decrease of retinal thickness at the macula with the resolution of cystoid macular edema. CONCLUSION: Resolution of diabetic macular edema with subsequent visual recovery is potentially associated with the vitreomacular separation in a patient after Nd:YAG laser capsulotomy. PMID- 10682990 TI - Reduced expression of the adherens junction protein cadherin-5 in a diabetic retina. AB - PURPOSE: Transvascular leakage occurs in diabetic retinopathy. The tight junction proteins occludin and zonula occludens-1 (ZO-1) and adherens junction protein cadherin-5 are critical to the maintenance of endothelial barrier. We report a comparison of junction protein expression in the normal and diabetic retina. METHOD: Case report. Postmortem retinal cryosections were prepared from the left eye of a 73-year-old woman with diabetic retinopathy. Cryosections were immunostained for cadherin-5, occludin, and ZO-1 and compared with retinal cryosections from the right eye of a 72-year-old man with no progression of retinal disease. RESULTS: Immunofluorescence showed positive retinal vessel staining for occludin and ZO-1 in both eyes and cadherin-5 in the normal eye but reduced cadherin-5 staining in the retinal vessels of the diabetic eye. CONCLUSION: Increases in transvascular leakage observed in diabetic retinal vasculature may be associated with reduction in the expression of the critical adherens junction protein, cadherin-5. PMID- 10682991 TI - Retinal vasculitis occurring with common variable immunodeficiency syndrome. AB - PURPOSE: To report severe retinal vasculitis causing decreased vision in three patients with the common variable immunodeficiency syndrome. METHOD: Case report. Three patients with common variable immunodeficiency syndrome developed decreased vision secondary to retinal vasculitis. Fluorescein angiography was performed in all three patients. Peribulbar injections were given in one patient, and two patients were treated with oral steroids and cyclosporin. RESULTS: All three patients were young and had classic common variable immunodeficiency syndrome. Bilateral retinal vasculitis and diffuse retinal edema were present in all three patients, and two patients had retinal neovascularization in the absence of ischemia. No evidence of intraocular infection was present, and none was detected systematically. Visual acuity decreased in five of the six eyes and was responsive to treatment in only one patient (both eyes). CONCLUSION: Retinal vasculitis may be another autoimmune manifestation of common variable immunodeficiency syndrome. PMID- 10682992 TI - Latanoprost increases the severity and recurrence of herpetic keratitis in the rabbit; latanoprost and herpes simplex keratitis. PMID- 10682993 TI - Epidemic and pandemic 'flu. PMID- 10682994 TI - Benefits of irradiation for DCIS: a Pyrrhic victory. PMID- 10682995 TI - Endpoints for homocysteine-lowering trials. PMID- 10682996 TI - Serum homocysteine and risk of coronary heart disease in UK Indian Asians. PMID- 10682997 TI - Neonatal screening for hearing impairment. PMID- 10682998 TI - Double gloving--electrical resistance and surgeons' resistance. PMID- 10682999 TI - Value of 6-min-walk test for assessment of severity and prognosis of heart failure. PMID- 10683000 TI - Effect of homocysteine-lowering treatment with folic acid plus vitamin B6 on progression of subclinical atherosclerosis: a randomised, placebo-controlled trial. AB - BACKGROUND: A high plasma homocysteine concentration is associated with increased risk of atherothrombotic disease. We investigated the effects of homocysteine lowering treatment (folic acid plus vitamin B6) on markers of subclinical atherosclerosis among healthy siblings of patients with premature atherothrombotic disease. METHODS: We did a randomised, placebo-controlled trial among 158 healthy siblings of 167 patients with premature atherothrombotic disease. 80 were assigned placebo and 78 were assigned 5 mg folic acid and 250 mg vitamin B6 daily for 2 years. The primary endpoint was the development or progression of subclinical atherosclerosis as estimated from exercise electrocardiography, the ankle-brachial pressure index, and carotid and femoral ultrasonography. FINDINGS: Ten participants in the treatment group, and 14 in the placebo group dropped out. Vitamin treatment, compared with placebo, was associated with a decrease in fasting homocysteine concentration (from 14.7 to 7.4 micromol/L vs from 14.7 to 12.0 micromol/L), and in postmethionine homocysteine concentration (from 64.9 to 34.9 micromol/L vs from 64.8 to 50.3 micromol/L). It was also associated with a decreased rate of abnormal exercise electrocardiography tests (odds ratio 0.40 [0.17-0.93]; p=0.035). There was no apparent effect of vitamin treatment on ankle-brachial pressure indices (0.87 [0.56-1.33]), or on carotid and peripheral-arterial outcome variables (1.02 [0.26 4.05] and 0.86 [0.47-1.59], respectively). INTERPRETATION: Homocysteine-lowering treatment with folic acid plus vitamin B6 in healthy siblings of patients with premature atherothrombotic disease is associated with a decreased occurrence of abnormal exercise electrocardiography tests, which is consistent with a decreased risk of atherosclerotic coronary events. PMID- 10683001 TI - Plasma homocysteine concentrations and risk of coronary heart disease in UK Indian Asian and European men. AB - BACKGROUND: Reasons for the increase in mortality due to coronary heart disease (CHD) in UK Indian Asians are not well understood. In this study, we tested the hypotheses that elevated plasma homocysteine concentrations are a risk factor for CHD in Indian Asians, and explain part of their increased CHD risk, compared with Europeans. METHODS: We undertook two parallel case-control studies, one in Europeans and one in Indian Asians. We recruited 551 male cases (294 European, 257 Indian Asian) and 1025 healthy male controls (507 European, 518 Indian Asian). Fasting and post-methionine load homocysteine, vitamin B12 and folate concentrations, and conventional CHD risk factors were measured. FINDINGS: Fasting homocysteine concentrations were 8% higher (95% CI 3-14) in cases compared with controls, in both ethnic groups. The odds ratio of CHD for a 5 micromol/L increment in fasting plasma homocysteine was 1.3 (1.1-1.6) in Europeans and 1.2 (1.0-1.4) in Indian Asians. The association between fasting plasma homocysteine and CHD was independent of conventional CHD risk factors in both ethnic groups. Post-load homocysteine concentrations were not significantly different in cases compared with controls. Among the controls, fasting homocysteine concentrations were 6% (2-10) higher in Indian Asians than in Europeans. From the results we estimate that elevated homocysteine may contribute to twice as many CHD deaths in Indian Asians, compared with Europeans. The differences in homocysteine concentrations between the two ethnic groups were explained by lower vitamin B12 and folate levels in Asians. INTERPRETATION: Plasma homocysteine is a novel and independent risk factor for CHD in Indian Asians, and may contribute to their increased CHD risk. Raised homocysteine concentrations in Indian Asians may be related to their reduced vitamin B12 and folate levels, implying that the increased CHD risk in this group may be reduced by dietary vitamin supplementation. PMID- 10683002 TI - Radiotherapy in breast-conserving treatment for ductal carcinoma in situ: first results of the EORTC randomised phase III trial 10853. EORTC Breast Cancer Cooperative Group and EORTC Radiotherapy Group. AB - BACKGROUND: Ductal carcinoma in situ (DCIS) of the breast is a disorder that has become more common since it may manifest as microcalcifications that can be detected by screening mammography. Since selected women with invasive cancer can be treated safely with breast conservation therapy it is paradoxical that total mastectomy has remained the standard treatment for DCIS. We did a randomised phase III clinical trial to investigate the role of radiotherapy after complete local excision of DCIS. METHODS: Between 1986 and 1996, women with clinically or mammographically detected DCIS measuring less than or equal to 5 cm were treated by complete local excision of the lesion and then randomly assigned to either no further treatment (n=503) or to radiotherapy (n=507; 50 Gy in 5 weeks to the whole breast). The median duration of follow-up was 4.25 years (maximum 12.0 years). All analyses were by intention to treat. FINDINGS: 500 patients were followed up in the no further treatment group and 502 in the radiotherapy group. In the no further treatment group 83 women had local recurrence (44 recurrences of DCIS, and 40 invasive breast cancer). In the radiotherapy group 53 women had local recurrences (29 recurrences of DCIS, and 24 invasive breast cancer). The 4 year local relapse-free was 84% in the group treated with local excision alone compared with 91% in the women treated by local excision plus radiotherapy (log rank p=0.005; hazard ratio 0.62). Similar reductions in the risk of invasive (40%, p=0.04) and non-invasive (35%, p=0.06) local recurrence were seen. CONCLUSIONS: Radiotherapy after local excision for DCIS, as compared with local excision alone, reduced the overall number of both invasive and non-invasive recurrences in the ipsilateral breast at a median follow-up of 4.25 years. PMID- 10683003 TI - Stress and psychiatric disorder in healthcare professionals and hospital staff. AB - BACKGROUND: Previous studies of stress in healthcare staff have indicated a probable high prevalence of distress. Whether this distress can be attributed to the stressful nature of the work situation is not clear. No previous study has used a detailed interview method to ascertain the link between stress in and outside of work and anxiety and depressive disorders. METHODS: Doctors, nurses, and administrative and ancillary staff were screened using the general health questionnaire (GHQ). High scorers (GHQ>4) and matched individuals with low GHQ scores were interviewed by means of the clinical interview schedule to ascertain definite anxiety and depressive disorders (cases). Cases and controls, matched for age, sex, and occupational group were interviewed with the life events and difficulties schedule classification and an objective measure of work stress to find out the amount of stress at work and outside of work. Sociodemographic and stress variables were entered into a logistic-regression analysis to find out the variables associated with anxiety and depressive disorders. FINDINGS: 64 cases and 64 controls were matched. Cases and controls did not differ on demographic variables but cases were less likely to have a confidant (odds ratio 0.09 [95% CI 0.01-0.79]) and more likely to have had a previous episode of psychiatric disorder (3.07 [1.10-8.57]). Cases and controls worked similar hours and had similar responsibility but cases had a greater number of objective stressful situations both in and out of work (severe event or substantial difficulty in and out of work-45 cases vs 18 controls 6.05 [2.81-13.00], p<0.001; severe chronic difficulty outside of work-27 vs 8, 5.12 [2.09-12.46], p<0.001). Cases had significantly more objective work problems than controls (median 6 vs 4, z=3.81, p<0.001). The logistic-regression analyses indicated that even after the effects of personal vulnerability to psychiatric disorder and ongoing social stress outside of work had been taken into account, stressful situations at work contributed to anxiety and depressive disorders. INTERPRETATION: Both stress at work and outside of work contribute to the anxiety and depressive disorders experienced by healthcare staff. Our findings suggest that the best way to decrease the prevalence of these disorders is individual treatment, which may focus on personal difficulties outside of work, combined with organisational attempts to reduce work stress. The latter may involve more assistance for staff who have a conflict between their managerial role and clinical role. PMID- 10683004 TI - Effects of replicating primary-reflex movements on specific reading difficulties in children: a randomised, double-blind, controlled trial. AB - BACKGROUND: Children with specific reading difficulties have problems that extend beyond the range of underlying language-related deficits (eg, they have difficulties with balance and motor control). We investigated the role of persistent primary reflexes (which are closely linked in the earliest months of life to the balance system) in disrupting the development of reading skills. METHODS: We assessed the efficacy of an intervention programme based on replicating the movements generated by the primary-reflex system during fetal and neonatal life. A randomised, individually matched, double-blind, placebo controlled design was used and children (aged 8-11 years) with persistent primary reflexes and a poor standard of reading were enrolled into one of three treatment groups: experimental (children were given a specific movement sequence); placebo control (children were given non-specific movements); and control (no movements). FINDINGS: From an initial sample of 98 children, 60 children, 20 in each group were matched on age, sex, verbal intelligence quotient (IQ), reading ability, and persistent asymmetrical tonic neck reflex. For asymmetrical tonic neck-reflex levels there was a significant (group by time) interaction (p<0.001). The experimental group showed a significant decrease in the level of persistent reflex over the course of the study (mean change -1.8 [95% CI -2.4 to -1.2], p<0.001), whereas the changes in the placebo-control and control groups were not significant (-0.2 [-0.9 to 0.6] and -0.4 [-0.9 to 0.2]). INTERPRETATION: This study provides further evidence of a link between reading difficulties and control of movement in children. In particular, our study highlights how the educational functioning of children may be linked to interference from an early neurodevelopmental system (the primary-reflex system). A new approach to the treatment of children with reading difficulties is proposed involving assessment of underlying neurological functioning, and appropriate remediation. PMID- 10683005 TI - Suppression and recovery of adrenal response after short-term, high-dose glucocorticoid treatment. AB - BACKGROUND: Suppression of the adrenal response is an unpredictable consequence of glucocorticoid treatment. To investigate the kinetics of the adrenal response after short-term, high-dose glucocorticoid treatment, we measured the adrenal response to the low-dose (1 microg) corticotropin stimulation test. METHODS: We studied 75 patients who received the equivalent of at least 25 mg prednisone daily for between 5 days and 30 days. After discontinuation of glucocorticoid treatment, 1 microg corticotropin was administered intravenously, and stimulated plasma cortisol concentrations were measured 30 min later. In patients with a suppressed response to 1 microg corticotropin, the test was repeated until stimulated plasma cortisol concentrations reached the normal range. FINDINGS: The adrenal response to 1 microg corticotropin was suppressed in 34 patients and normal in 41. Subsequent low-dose corticotropin tests showed a steady recovery of the adrenal response within 14 days. In two patients, the adrenal response remained suppressed for several months. There was no correlation between plasma cortisol concentrations and the duration or dose of glucocorticoid treatment. INTERPRETATION: Suppression of the adrenal response is common after short-term, high-dose glucocorticoid treatment. The low-dose corticotropin test is a sensitive and simple test to assess the adrenal response after such treatment. PMID- 10683006 TI - A woman with ascites and abdominal masses. PMID- 10683007 TI - Indinavir concentrations and St John's wort. AB - St John's wort reduced the area under the curve of the HIV-1 protease inhibitor indinavir by a mean of 57% (SD 19) and decreased the extrapolated 8-h indinavir trough by 81% (16) in healthy volunteers. A reduction in indinavir exposure of this magnitude could lead to the development of drug resistance and treatment failure. PMID- 10683008 TI - Acute heart transplant rejection due to Saint John's wort. AB - We report here acute rejection in two transplant patients due to a metabolic interaction of St John's wort and cyclosporin. PMID- 10683009 TI - School-based hepatitis B vaccination programme and adolescent multiple sclerosis. AB - We investigated multiple sclerosis in adolescents in British Columbia before and after a hepatitis B vaccination programme was begun. There was no evidence of a link between hepatitis B vaccination and multiple sclerosis or other demyelinating disease. PMID- 10683010 TI - Pulse-inversion contrast harmonic imaging: ultrasonographic assessment of cerebral perfusion. AB - Pulse-inversion contrast harmonic imaging is a new ultrasonographic technique that can assess brain perfusion. In an adult with moyamoya disease and multiple recurrent strokes, this method detected subtle hemispheric differences in temporal-lobe perfusion, presumably due to neovascularisation, which were not shown by xenon-computed tomography or magnetic resonance perfusion imaging. PMID- 10683011 TI - Simian foamy virus infection among zoo keepers. AB - We investigated 322 North American zoo workers in an anonymous serosurvey for antibodies to simian foamy viruses to establish the potential risk of zoonotic transmission by these retroviruses. 4 of 133 (3%) individuals who worked specifically with mammals including primates were seropositive, primarily with chimp-like viruses, indicating the importance of work practices to reduce exposure to these agents. PMID- 10683012 TI - Suspicions raised over breast-cancer-therapy trial. PMID- 10683013 TI - Functional foods nibble away at serum cholesterol concentrations. PMID- 10683014 TI - Experts hear sobering side of HIV-1 treatment story. PMID- 10683015 TI - Epidemiological studies: overdone or underappreciated? PMID- 10683016 TI - British American tobacco shrugs off smuggling charges. PMID- 10683017 TI - New Zealand's pharmaceutical reference-pricing strategy may backfire. PMID- 10683018 TI - South Africa's AIDS taskforce questioned. PMID- 10683019 TI - Are booster immunisations needed for lifelong hepatitis B immunity? European Consensus Group on Hepatitis B Immunity. AB - Long-term protection against clinically significant breakthrough hepatitis B (HB) virus infection and chronic carriage depends on immunological memory, which allows a protective anamnestic antibody response to antigen challenge. Memory seems to last for at least 15 years in immunocompetent individuals. To date there are no data to support the need for booster doses of HB vaccine in immunocompetent individuals who have responded to a primary course. All adequately vaccinated individuals have shown evidence of immunity in the form of persisting anti-HBs and/or in vitro B-cell stimulation or an anamnestic response to a vaccine challenge. Nonetheless several countries and individuals currently have a policy of administering booster doses to certain risk groups. Boosters may be used to provide reassurance of protective immunity against benign breakthrough infection. For immunocompromised patients, regular testing for anti-HBs, and a booster injection when the titre falls below 10 mIU/mL, is advised. Long-term monitoring should continue, to confirm the absence of clinically significant breakthrough episodes of hepatitis B and to find out if a carrier state develops after 15 years. Also, non-responders to a primary course should continue to be studied. PMID- 10683020 TI - Value of drug-licensing documents in studying the effect of postmenopausal hormone therapy on cardiovascular disease. AB - BACKGROUND: In a previous study of pooled data from published trials, we found no evidence to support the claim that postmenopausal hormone therapy (PHT) is associated with a decrease in cardiovascular disease. The purpose of this study was to see whether reports of clinical trials attached to drug-licensing applications in Finland could be obtained for scientific purposes, whether they are useful for studying cardiovascular events resulting from PHT, and if so, whether these unpublished reports corroborate the results of published reports. METHODS: Since clinical trials in drug-licensing documents are confidential, we had to obtain special permission from the Ministry of Social Affairs and Health to use the data for research purposes. After permission was granted, we studied the clinical sections of licensing documents for PHT drugs sent by drug companies to the Finnish Drug Agency. We aimed to identify trials that compared PHT and a placebo (or no therapy, or vitamin-mineral drugs), and that reported on cardiovascular and thromboembolic events or superficial phlebitis. New trials were identified and their data were pooled with those of published trials. FINDINGS: 17 licensing applications for drugs used as PHT were found. The number, type, and quality of reporting of clinical trials varied widely between applications. The trials and their reporting of unanticipated adverse events were mostly inadequate. Six new trials (ie, those fulfilling the inclusion criteria and not included in our earlier report) were found. The new trials added little to the conclusions of previously published studies: the calculated odds ratios of cardiovascular and thromboembolic events for women taking PHT versus those not taking it was 1.97 (95% CI 0.84-4.63), compared with 1.65 (0.65-4.21) in our previous study. INTERPRETATION: In this case, unpublished trials added only a little to the data available from published trials, mainly due to the type of clinical data used in the licensing applications. The new data did not change the previous conclusion that clinical trials do not support a beneficial effect of PHT on cardiovascular diseases. PMID- 10683021 TI - The death of a British officer-cadet from heat illness. PMID- 10683022 TI - Management of unstable coronary-artery disease. PMID- 10683023 TI - Management of unstable coronary-artery disease. PMID- 10683024 TI - Management of unstable coronary-artery disease. PMID- 10683025 TI - Management of unstable coronary-artery disease. PMID- 10683026 TI - Nimesulide and neonatal renal failure. PMID- 10683027 TI - Safety of St John's wort (Hypericum perforatum) PMID- 10683028 TI - Safety of St John's wort (Hypericum perforatum) PMID- 10683029 TI - Safety of St John's wort (Hypericum perforatum) PMID- 10683030 TI - Safety of St John's wort (Hypericum perforatum) PMID- 10683031 TI - Taxane-induced glaucoma. PMID- 10683032 TI - Urinary incontinence and the rhabdosphincter. PMID- 10683033 TI - Hepatitis E virus infection. PMID- 10683034 TI - Pneumococcal vaccine and the elderly. PMID- 10683035 TI - End of exile for Taiwan. PMID- 10683036 TI - Early uptake of research findings after fast-track publication. PMID- 10683037 TI - Trade agreements and public health: role of WHO. PMID- 10683038 TI - Judgment by peers. PMID- 10683039 TI - The Nobel chronicles. 1987: Susumu Tonegawa (b 1939). PMID- 10683040 TI - Recent advances in wound healing. PMID- 10683041 TI - IRBs search for answers and support during a time of institutional change. PMID- 10683042 TI - Music therapists chime in with data on medical results. PMID- 10683043 TI - From the Centers for Disease Control and Prevention. Achievements in public health, 1900-1999: changes in the public health system. PMID- 10683044 TI - Effects of androstenedione in young men. PMID- 10683045 TI - Effects of androstenedione in young men. PMID- 10683046 TI - Effects of androstenedione in young men. PMID- 10683047 TI - Animal research and human disease. PMID- 10683049 TI - Chronic fatigue: syndrome or disease? PMID- 10683048 TI - Animal research and human disease. PMID- 10683050 TI - Alcohol consumption and risk of coronary heart disease. PMID- 10683051 TI - Immunologic strategies for herpes vaccination. PMID- 10683052 TI - Immunologic strategies for herpes vaccination. PMID- 10683053 TI - A controlled trial of a critical pathway for treatment of community-acquired pneumonia. CAPITAL Study Investigators. Community-Acquired Pneumonia Intervention Trial Assessing Levofloxacin. AB - CONTEXT: Large variations exist among hospitals in the use of treatment resources for community-acquired pneumonia (CAP). Lack of a common approach to the diagnosis and treatment of CAP has been cited as an explanation for these variations. OBJECTIVE: To determine if use of a critical pathway improves the efficiency of treatment for CAP without compromising the well-being of patients. DESIGN: Multicenter controlled clinical trial with cluster randomization and up to 6 weeks of follow-up. SETTING: Nineteen teaching and community hospitals in Canada. PATIENTS: A total of 1743 patients with CAP presenting to the emergency department at 1 of the participating institutions between January 1 and July 31, 1998. INTERVENTION: Hospitals were assigned to continue conventional management (n = 10) or implement the critical pathway (n = 9), which consisted of a clinical prediction rule to guide the admission decision, levofloxacin therapy, and practice guidelines. MAIN OUTCOME MEASURES: Effectiveness of the critical pathway, as measured by health-related quality of life on the Short-Form 36 Physical Component Summary (SF-36 PCS) scale at 6 weeks; and resource utilization, as measured by the number of bed days per patient managed (BDPM). RESULTS: Quality of life and the occurrence of complications, readmission, and mortality were not different for the 2 strategies; the 1-sided 95% confidence limit of the between-group difference in the SF-36 PCS change score was 2.4 points, which was within a predefined 3-point boundary for equivalence. Pathway use was associated with a 1.7-day reduction in BDPM (4.4 vs 6.1 days; P = .04) and an 18% decrease in the admission of low-risk patients (31% vs 49%; P = .01). Although inpatients at critical pathway hospitals had more severe disease, they required 1.7 fewer days of intravenous therapy (4.6 vs 6.3 days; P = .01) and were more likely to receive treatment with a single class of antibiotic (64% vs 27%; P<.001). CONCLUSION: In this study, implementation of a critical pathway reduced the use of institutional resources without causing adverse effects on the well-being of patients. PMID- 10683054 TI - Outcome at age 4 years in offspring of women with maternal phenylketonuria: the Maternal PKU Collaborative Study. AB - CONTEXT: Untreated maternal phenylketonuria (PKU) increases risk for developmental problems in offspring. The extent to which this risk is reduced by maternal dietary therapy at various stages of pregnancy is not known. OBJECTIVE: To determine whether dietary treatment during pregnancy of women with PKU affects developmental outcomes of offspring. DESIGN: The Maternal PKU Collaborative Study, an ongoing, longitudinal prospective study begun in 1984. SETTING: A total of 78 metabolic clinics and obstetrical offices in the United States, Canada, and Germany. PARTICIPANTS: A total of 253 children of women with PKU (n = 149), with untreated mild hyperphenylalaninemia (n = 33), or without known metabolic problems (comparison group; n = 71) were followed up to age 4 years. INTERVENTION: Women with PKU were offered a low-phenylalanine diet prior to or during pregnancy with the aim of maintaining metabolic control (plasma phenylalanine < or =10 mg/dL [< or =605 micromol/L]). Women with mild hyperphenylalaninemia, who had plasma phenylalanine levels of no more than 10 mg/dL (605 micromol/L) on a normal diet, were not treated. MAIN OUTCOME MEASURES: Children's scores on cognitive and behavioral assessments (McCarthy Scales of Children's Abilities, Test of Language Development, Achenbach Child Behavior Checklist, Vineland Adaptive Behavior Scales, and Home Observation for Measurement of the Environment), compared by maternal metabolic status at 0 to 10 weeks', 10 to 20 weeks', and after 20 weeks' gestation. RESULTS: Scores on the McCarthy General Cognitive Index decreased as weeks to metabolic control increased (r = -0.58; P<.001). Offspring of women who had metabolic control prior to pregnancy had a mean (SD) score of 99 (13). Forty-seven percent of offspring whose mothers did not have metabolic control by 20 weeks' gestation had a General Cognitive Index score 2 SDs below the norm. Overall, 30% of children born to mothers with PKU had social and behavioral problems. CONCLUSIONS: Our data suggest that delayed development in offspring of women with PKU is associated with lack of maternal metabolic control prior to or early in pregnancy. Treatment at any time during pregnancy may reduce the severity of delay. PMID- 10683056 TI - End-of-life care content in 50 textbooks from multiple specialties. AB - CONTEXT: Prior reviews of small numbers of medical textbooks suggest that end-of life care is not well covered in textbooks. No broad study of end-of-life care content analysis has been performed on textbooks across a wide range of medical, pediatric, psychiatric, and surgical specialties. OBJECTIVE: To determine the quantity and rate the adequacy of information on end-of-life care in textbooks from multiple medical disciplines. DESIGN AND SOURCES: A 1998 review of 50 top selling textbooks from multiple specialties (cardiology, emergency medicine, family and primary care medicine, geriatrics, infectious disease and acquired immunodeficiency syndrome [AIDS], internal medicine, neurology, oncology and hematology, pediatrics, psychiatry, pulmonary medicine, and surgery) for the presence and adequacy of content in 13 end-of-life care domains. MAIN OUTCOME MEASURES: Chapters on diseases commonly causing death and those devoted to end-of life care were identified, read, rated, and compared by textbook specialty, chapter, and domain for the presence of helpful information in the 13 domains. Content for each domain was rated as absent, minimally present, or helpful. Textbook indexes were analyzed for the number of pages relevant to end-of-life care. RESULTS: Overall, helpful information was provided in 24.1% (range, 8.7% 44.2%) of the expected end-of-life content domains; in 19.1% (range, 6.2%-38.5%), expected content received minimal attention; and in 56.9% (range, 23.1 %-77.9%), expected content was absent. As a group, the textbooks with the highest percentages of absent content were in surgery (71.8%), infectious diseases and AIDS (70%), and oncology and hematology (61.9%). Textbooks with the highest percentage of helpful end-of-life care content were in family medicine (34.4%), geriatrics (34.4%), and psychiatry (29.6%). In internal medicine textbooks, the content domains with the greatest amount of helpful information were epidemiology and natural history. Content domains covered least well were social, spiritual, ethical, and family issues, as well as physician after-death responsibilities. On average, textbook indexes cited 2% of their total pages as pertinent to end-of life care. CONCLUSION: Top-selling textbooks generally offered little helpful information on caring for patients at the end of life. Most disease-oriented chapters had no or minimal end-of-life care content. Specialty textbooks with information about particular diseases often did not contain helpful information on caring for patients dying from those diseases. PMID- 10683055 TI - Testosterone replacement and resistance exercise in HIV-infected men with weight loss and low testosterone levels. AB - CONTEXT: Previous studies of testosterone supplementation in HIV-infected men failed to demonstrate improvement in muscle strength. The effects of resistance exercise combined with testosterone supplementation in HIV-infected men are unknown. OBJECTIVE: To determine the effects of testosterone replacement with and without resistance exercise on muscle strength and body composition in HIV infected men with low testosterone levels and weight loss. DESIGN AND SETTING: Placebo-controlled, double-blind, randomized clinical trial conducted from September 1995 to July 1998 at a general clinical research center. PARTICIPANTS: Sixty-one HIV-infected men aged 18 to 50 years with serum testosterone levels of less than 12.1 nmol/L (349 ng/dL) and weight loss of 5% or more in the previous 6 months, 49 of whom completed the study. INTERVENTIONS: Participants were randomly assigned to 1 of 4 groups: placebo, no exercise (n = 14); testosterone enanthate (100 mg/wk intramuscularly), no exercise (n = 17); placebo and exercise (n = 15); or testosterone and exercise (n = 15). Treatment duration was 16 weeks. MAIN OUTCOME MEASURES: Changes in muscle strength, body weight, thigh muscle volume, and lean body mass compared among the 4 treatment groups. RESULTS: Body weight increased significantly by 2.6 kg (P<.001) in men receiving testosterone alone and by 2.2 kg (P = .02) in men who exercised alone but did not change in men receiving placebo alone (-0.5 kg; P = .55) or testosterone and exercise (0.7 kg; P = .08). Men treated with testosterone alone, exercise alone, or both experienced significant increases in maximum voluntary muscle strength in leg press (range, 22%-30%), leg curls (range, 18%-36%), bench press (range, 19%-33%), and latissimus pulls (range, 17%-33%). Gains in strength in all exercise categories were greater in men assigned to the testosterone-exercise group or to the exercise-alone group than in those assigned to the placebo-alone group. There was a greater increase in thigh muscle volume in men receiving testosterone alone (mean change, 40 cm3; P<.001 vs zero change) or exercise alone (62 cm3; P = .003) than in men receiving placebo alone (5 cm3; P = .70). Average lean body mass increased by 2.3 kg (P = .004) and 2.6 kg (P<.001), respectively, in men who received testosterone alone or testosterone and exercise but did not change in men receiving placebo alone (0.9 kg; P = .21). Hemoglobin levels increased in men receiving testosterone but not in those receiving placebo. CONCLUSION: Our data suggest that testosterone and resistance exercise promote gains in body weight, muscle mass, muscle strength, and lean body mass in HIV-infected men with weight loss and low testosterone levels. Testosterone and exercise together did not produce greater gains than either intervention alone. PMID- 10683057 TI - Oral androstenedione administration and serum testosterone concentrations in young men. AB - CONTEXT: Androstenedione, a steroid hormone and the major precursor to testosterone, is available without prescription and is purported to increase strength and athletic performance. The hormonal effects of androstenedione, however, are unknown. OBJECTIVE: To determine if oral administration of androstenedione increases serum testosterone levels in healthy men. DESIGN: Open label randomized controlled trial conducted between October 1998 and April 1999. SETTING: General clinical research center of a tertiary-care, university affiliated hospital. PARTICIPANTS: Forty-two healthy men aged 20 to 40 years. INTERVENTION: Subjects were randomized to receive oral androstenedione (either 100 mg/d [n = 15] or 300 mg/d [n = 14]) or no androstenedione (n = 13) for 7 days. MAIN OUTCOME MEASURES: Changes in serum testosterone, androstenedione, estrone, and estradiol levels, measured by frequent blood sampling, compared among the 3 treatment groups. RESULTS: Mean (SE) changes in the area under the curve (AUC) for serum testosterone concentrations were -2% (7%), -4% (4%), and 34% (14%) in the groups receiving 0, 100, and 300 mg/d of androstenedione, respectively. When compared with the control group, the change in testosterone AUC was significant for the 300-mg/d group (P<.001) but not for the 100-mg/d group (P = .48). Baseline testosterone levels, drawn 24 hours after androstenedione administration, did not change. Mean (SE) changes in the AUC for serum estradiol concentrations were 4% (6%), 42% (12%), and 128% (24%) in the groups receiving 0, 100, and 300 mg/d of androstenedione, respectively. When compared with the control group, the change in the estradiol AUC was significant for both the 300-mg/d (P<.001) and 100-mg/d (P = .002) groups. There was marked variability in individual responses for all measured sex steroids. CONCLUSIONS: Our data suggest that oral androstenedione, when given in dosages of 300 mg/d, increases serum testosterone and estradiol concentrations in some healthy men. PMID- 10683058 TI - Effect of out-of-hospital pediatric endotracheal intubation on survival and neurological outcome: a controlled clinical trial. AB - CONTEXT: Endotracheal intubation (ETI) is widely used for airway management of children in the out-of-hospital setting, despite a lack of controlled trials demonstrating a positive effect on survival or neurological outcome. OBJECTIVE: To compare the survival and neurological outcomes of pediatric patients treated with bag-valve-mask ventilation (BVM) with those of patients treated with BVM followed by ETI. DESIGN: Controlled clinical trial, in which patients were assigned to interventions by calendar day from March 15, 1994, through January 1, 1997. SETTING: Two large, urban, rapid-transport emergency medical services (EMS) systems. PARTICIPANTS: A total of 830 consecutive patients aged 12 years or younger or estimated to weigh less than 40 kg who required airway management; 820 were available for follow-up. INTERVENTIONS: Patients were assigned to receive either BVM (odd days; n = 410) or BVM followed by ETI (even days; n = 420). MAIN OUTCOME MEASURES: Survival to hospital discharge and neurological status at discharge from an acute care hospital compared by treatment group. RESULTS: There was no significant difference in survival between the BVM group (123/404 [30%]) and the ETI group (110/416 [26%]) (odds ratio [OR], 0.82; 95% confidence interval [CI], 0.61-1.11) or in the rate of achieving a good neurological outcome (BVM, 92/404 [23%] vs ETI, 85/416 [20%]) (OR, 0.87; 95% CI, 0.62-1.22). CONCLUSION: These results indicate that the addition of out-of-hospital ETI to a paramedic scope of practice that already includes BVM did not improve survival or neurological outcome of pediatric patients treated in an urban EMS system. PMID- 10683059 TI - Genital herpes and public health: addressing a global problem. AB - Genital herpes can be caused by herpes simplex virus 2 (HSV-2) or, less commonly, by herpes simplex virus 1 (HSV-1). With a seroprevalence of antibodies to HSV-2 of 22% in the general population, genital herpes is 1 of the 3 most prevalent sexually transmitted diseases (STDs) in the United States. A central issue in the public health problem of genital herpes is the high proportion of genital HSV infections that are unrecognized by both patients and clinicians. Persons who are HSV-2 seropositive may be symptomatic but nevertheless fail to recognize genital herpes; they serve as reservoirs for transmission. Physicians and patients must be aware of the subclinical presentation of genital herpes and the potential these patients have for transmitting HSV. Serious consequences of HSV infection include neonatal herpes and increased risk of human immunodeficiency virus transmission. Recommendations to physicians for prevention include using type specific tests for HSV when screening for other STDs and testing for HSV when evaluating patients with genital ulcers. Researchers must evaluate the performance of type-specific tests and strategies to prevent transmission. PMID- 10683060 TI - Threats to the confidentiality of medical records--no place to hide. PMID- 10683061 TI - Out-of-hospital intubation of children. PMID- 10683062 TI - Uses and abuses of prescription drug information in pharmacy benefits management programs. AB - A 1998 incident in which patients' prescription information was used to advertise a new drug exemplifies the importance of confidentiality in the era of managed care and computers. The ethical concerns voiced about this incident can also apply to pharmacy benefits management programs. The use of personal health information in pharmacy benefits management is particularly important because of increased pressures to control rising drug costs. Specific confidentiality concerns include whether the goal of benefiting patients will be achieved and whether the means are appropriate. The means may be problematic because of financial conflicts of interest, lack of patient authorization, inappropriate access to information by third parties, and inadequate safeguards for confidentiality. Policies should be crafted that protect confidentiality while allowing appropriate use of personal health information in pharmacy benefits management. Sound policies should require clear evidence of benefit to patients, an oversight committee, patient authorization, disclosure or prohibition of conflicts of interest, additional safeguards for sensitive medical conditions, strong confidentiality protections, and restrictions on advertising. PMID- 10683063 TI - JAMA Patient Page: pneumonia. PMID- 10683064 TI - Follicular carcinoma of the thyroid gland: prognostic factors, treatment, and survival. AB - Prognostic variables and treatment outcomes of 82 patients treated at the Northern Israel Oncology Center were reviewed. There were 59 women and 23 men in this series. The female/male ratio was 2.6/1. Median age was 46 years. Median follow-up was 11.4 (range: 3.8-24 years). Median tumor size was 3.6 cm. When first seen, 4 patients had lymph node involvement and 11 (13%) had distant metastases. Surgical treatment was total thyroidectomy in 37 patients (45%), subtotal thyroidectomy in 38 (46%), and lesser procedures in 7 (9%). Sixty-six patients (80%) were treated after surgery with 131I to ablate thyroid remnants. Doses ranged between 30 and 80 mCi. The 20-year overall actuarial survival rate was 65%. The actuarial survival rate of patients <40 years of age was 96% versus 33% in patients >50 years of age (p = 0.0008). Patients with distant metastases at presentation had inferior survival compared with patients without metastases. In conclusion, we found subtotal thyroidectomy followed by 131I and hormone therapy to provide survival similar to that with total thyroidectomy, with less morbidity. Risk factors include: age > or =40 at the time of diagnosis, presence of distant metastases, capsular invasion, tumor size > or =2 cm, and male gender. PMID- 10683065 TI - Comparison of preoperative embolization followed by radical nephrectomy with radical nephrectomy alone for renal cell carcinoma. AB - A series of 474 patients with renal cell carcinoma (RCC), who had radical nephrectomy during a period of 15 years, was studied to assess the prognostic significance of various pathologic parameters (tumor stage [pT], lymph node status, metastasis, tumor grade, venous involvement) and value of preoperative embolization of renal artery. There were: 20 (4%) pT1, 204 (43%) pT2, 245 (52%) pT3, and 5 (1%) pT4 patients. All 474 patients underwent nephrectomy including a group of 118 (25%) patients (24 pT2, 90 pT3, and 4 pT4) who underwent preoperative embolization of the renal artery. To compare treatment outcomes in embolized patients with RCC, a group of 116 (24%) nonembolized patients with RCC was selected. This group was matched for sex, age, stage, tumor size, and tumor grade, with the embolized patients (p<0.01). All important prognostic factors were studied as to their influence on survival by the treatment group. The overall 5- and 10-year survival was 62% and 47%, respectively. The 5- and 10-year survival rates were significantly better (p<0.01) for patients with pT2 than for those with pT3 tumors (79% vs. 50% and 59% vs. 35%, respectively). Involvement of regional lymph nodes (N+) was an important prognostic factor for survival in patients with pT3 tumors. The 5-year survival for pT3 N+ was 39%, compared with 66% in those with pT3N0 (p<0.01). Preoperative embolization was also an important factor influencing survival. The overall 5- and 10-year survival for 118 patients embolized before nephrectomy was 62% and 47%, respectively, and it was 35% and 23%, respectively, for the matched group of 116 patients treated with surgery alone (p = 0.01). The most important finding of this study was an apparent importance of preoperative embolization in improving patients' survival. This finding needs to be interpreted with caution and confirmed in a prospective randomized trial. PMID- 10683066 TI - A phase II trial of tamoxifen, ifosfamide, epirubicin, and cisplatin combination chemotherapy for inoperable non-small-cell lung cancer. AB - A phase II trial of tamoxifen, ifosfamide, epirubicin, and cisplatin (TIEP) chemotherapy was conducted in patients with chemonaive inoperable non-small-cell lung cancer (NSCLC) to assess response and toxicity. From October 1997 to August 1998, 19 patients were treated. The treatment schema included tamoxifen 60 mg twice daily by mouth on days 1 to 3, ifosfamide 3 g/m2 intravenous infusion (IV) 60 minutes with mesna on day 2, epirubicin 50 mg/m2 IV bolus on day 2, and cisplatin 60 mg/m2 IV 60 minutes on day 2 every 4 weeks for up to six cycles. All patients were evaluable for response and toxicity. The major toxicity was myelosuppression; grade 3 or 4 leukopenia or neutropenia occurred in 14 of 19 (73.7%) patients during treatment, and 6 patients (31.6%) experienced fever in association with the neutropenia; no toxic deaths occurred. Grade 3 anemia occurred in six patients (31.6%) during the treatment. Grade 3 or 4 nausea/vomiting occurred in only one patient. Toxicities other than neutropenia and anemia were minimal. After two cycles of treatment, 9 of 19 patients attained a partial response (47.4%, 95% confidence interval 24.9%-69.9%) in this study. The median time to disease progression was 6 months and median survival time was 12 months. We conclude that TIEP is an active combination regimen with an acceptable toxicity profile in Chinese patients with inoperable NSCLC. PMID- 10683067 TI - Late pulmonary effects in favorable stage I and IIA Hodgkin's disease treated with radiotherapy alone. AB - Radiotherapy (RT) in patients with favorable-stage Hodgkin's disease can induce clinical and subclinical evidence of pulmonary damage lasting over the years. In this study, we monitored 36 patients with stage IA-IIA Hodgkin's disease treated with subtotal nodal RT. The planned dose of RT was 40 Gy to 44 Gy to the involved areas and 36 Gy to the adjacent uninvolved areas. Pulmonary function was evaluated by chest radiograph, spirometric parameters, arterial blood gas analysis, and single-breath CO transfer factor (DLCO). The tests were performed before and at the end of irradiation, and during the follow-up 1 and 3 to 5 years after the treatment. At the end of RT, we found a significant decrease of total lung capacity, vital capacity, forced expiratory volume in 1 second, residual volume, and DLCO. Spirometric parameters improved during the follow-up period, whereas the decline of DLCO (-6.4%) was persistent. No correlation was found between mantle RT dose and DLCO changes. Four patients showed a decline of DLCO of >20% from pretreatment values but only one was symptomatic. Our study confirms that RT induces a pulmonary-restrictive disease at a subclinical level that seems to be reversible in the majority of patients. PMID- 10683068 TI - Ambamustine in the second-line treatment of patients with small-cell lung cancer: a phase II Fonicap study. AB - Despite a high probability of response to first-line chemotherapy, most patients with small-cell lung cancer (SCLC) will eventually have progression of their disease because of the development of resistant disease. Second-line testing of new drugs is an accepted research strategy in SCLC. In this context, the Italian Lung Cancer Task Force (FONICAP) has tested a new synthetic bifunctional alkylating agent, Ambamustine, with preliminary evidence of activity in other solid tumors. Patients with measurable SCLC, progressive after one first-line chemotherapy regimen (either "sensitive" or "refractory"), were eligible for the study. Ambamustine was administered at the dose of 2 mg/kg as a 1-hour intravenous infusion on day 1 every 21 days. The dose was to be increased to 3 mg/kg if no grade IV toxicity and complete hematologic recovery had occurred by day 22. Sample size was calculated according to a two-stage optimal Simon's design. Seventeen patients were entered into the study. Twelve patients were refractory to prior chemotherapy; 12 had extensive disease; the median age was 64 years (range: 46-75 years) and the median performance status was 1. Among 13 patients who received more than one cycle, 9 patients could increase Ambamustine dose from 2 to 3 mg/kg. No objective response was observed: one patient obtained a 50% regression of the primary tumor with contemporary disease progression in the liver and was qualified as having progressive disease. The treatment was well tolerated: grade IV leukopenia occurred in only 1 patient; grade III anemia occurred in 17.6%, grade III leukopenia in 11.8%, and grade III thrombocytopenia in 23.5%. Nonhematologic toxicity was minimal. Ambamustine, at the dose and schedule used in this study, is well tolerated in pretreated patients with SCLC but has no significant antitumor activity in this unfavorable group of patients. PMID- 10683069 TI - Kaposi sarcoma after treatment of Hodgkin's disease in a young adult non-AIDS patient: case report and review. AB - We describe a young woman diagnosed with Hodgkin's disease, stage I, at age 20 years. She delayed treatment until age 23, at which time she was considered to have stage II-A disease and was then treated with chemotherapy and involved field irradiation. Two years later, Kaposi sarcoma, which developed on her right shoulder, was excised. Both the Hodgkin's disease and Kaposi sarcoma appeared to be cured, but 3 years later, acute myelogenous leukemia developed and the patient subsequently died in relapse. This is one of the very few instances of a young patient, not infected with the AIDS virus, in whom Kaposi sarcoma developed as a second malignancy after treatment of Hodgkin's disease. PMID- 10683070 TI - 'Full dose' reirradiation of human cervical spinal cord. AB - With the progress of modern multimodality cancer treatment, retreatment of late recurrences or second tumors became more commonly encountered in management of patients with cancer. Spinal cord retreatment with radiation is a common problem in this regard. Because radiation myelopathy may result in functional deficits, many oncologists are concerned about radiation-induced myelopathy when retreating tumors located within or immediately adjacent to the previous radiation portal. The treatment decision is complicated because it requires a pertinent assessment of prognostic factors with and without reirradiation, radiobiologic estimation of recovery of occult spinal cord damage from the previous treatment, as well as interactions because of multimodality treatment. Recent studies regarding reirradiation of spinal cord in animals using limb paralysis as an endpoint have shown substantial and almost complete recovery of spinal cord injury after a sufficient time after the initial radiotherapy. We report a case of "full" dose reirradiation of the entire cervical spinal cord in a patient who has not developed clinically detectable radiation-induced myelopathy on long-term follow up of 17 years after the first radiotherapy and 5 years after the second radiotherapy. PMID- 10683071 TI - Thoracic wall prosthesis prevents deep invasion by non-small-cell lung cancer. AB - Chest wall invasion is found in 5% of patients with non-small-cell lung cancer. Treatment for localized non-small-cell lung cancer consists of surgical resection and/or radiotherapy. We report a patient with lung cancer who had a local relapse after a reconstruction of the thoracic wall with a soft-tissue patch. Chemotherapy was given before reresection of the local relapse. Postoperative radiation therapy was performed. Twenty-one months after treatment for recurrent disease, the patient remains in complete remission. The history of this patient shows that a soft-tissue patch may prevent local tumor invasion. A review of the literature is given. PMID- 10683072 TI - Sustained ventricular tachycardia and its successful prophylaxis during high-dose bolus interleukin-2 therapy for metastatic renal cell carcinoma. AB - In the setting of interleukin-2 (IL-2) administration, tachycardias of ventricular origin are classified as serious, grade IV toxicities, necessitating the discontinuation of therapy. In this report, we describe a patient with renal cell carcinoma who experienced ventricular tachycardia while undergoing treatment with high-dose bolus IL-2. Prophylaxis with sotalol permitted the successful completion of his first cycle of treatment, without any recurrent rhythm disturbances. PMID- 10683073 TI - A phase II trial of 5-fluorouracil, leucovorin, and interferon alpha 2A (IFN alpha 2a) in metastatic pancreatic carcinoma: a Penn Cancer Clinical Trials Group (PCCTG) trial. AB - A phase II study was performed to evaluate the activity and toxicity of 5 fluorouracil (5-FU), leucovorin, and inteferon alpha-2a in metastatic pancreatic carcinoma. Twenty-three patients were entered in this study. Four patients withdrew before receiving treatment and one patient was nonevaluable for response because of treatment-related toxicity. The most common significant toxicity was nausea and vomiting. Treatment-related hospitalization was significant. Of 18 evaluable patients, 4 maintained stable disease and 14 had disease progression. None had an objective clinical response. We conclude that this biochemically modulated 5-FU regimen is ineffective treatment for advanced pancreatic carcinoma, with significant toxicity even in highly selected patients with an ambulatory performance status. PMID- 10683074 TI - Continuous-infusion high-dose leucovorin with 5-fluorouracil and cisplatin for relapsed metastatic breast cancer: a phase II study. AB - Twelve women with metastatic breast cancer were treated with continuous infusion high dose leucovorin, 5-fluorouracil and cisplatin. Toxicity was severe although the dose was lower than previously described for the treatment of other cancers, and there was little anti-tumor activity. Many other regimens are more effective and less toxic. PMID- 10683075 TI - Pyomyositis after chemotherapy for breast cancer. AB - Pyomyositis is a rare complication of chemotherapy. A 47-year-old woman with metastatic breast cancer, in whom pyomyositis developed after chemotherapy, is described. It was difficult to differentiate between pyomyositis and deep venous thrombosis early in her admission. Pyomyositis should be considered part of the differential diagnosis of deep venous thrombosis. This infection, after chemotherapy, usually is considered to be caused by neutropenia or immunodeficiency secondary to the cancer, or both. It is postulated that subclinical myopathy, secondary to the malignancy or drugs used in treating the malignancy, or both, may also predispose to pyomyositis. PMID- 10683076 TI - Treatment of squamous cell esophageal cancer with topotecan: an Eastern Cooperative Oncology Group Study (E2293). AB - Seventeen patients with enhanced measurable squamous cell carcinoma of the esophagus were treated with topotecan 1.5 mg/m2 daily for 5 days repeated every 21 days. Toxicity was severe, with 1 death from myelotoxicity and 10 patients with life-threatening myelotoxicity. Severe gastrointestinal toxicity consisting of vomiting was also seen in three patients. No response was seen in any of the patients in the study. Topotecan given in this manner has no activity in squamous cell carcinoma of the esophagus. PMID- 10683077 TI - Accuracy and clinical impact of mediastinal lymph node staging with FDG-PET imaging in potentially resectable lung cancer. AB - To determine the sensitivity, specificity, and accuracy of staging mediastinal nodal disease in potentially resectable lung cancer using fluorodeoxyglucose positron emission tomography (FDG-PET), computed tomography (CT), or both and compare these results to surgical staging. We also assessed whether PET scanning results changed clinical management. From 1992 to 1997, 50 patients underwent CT, and PET scanning before or close to the time of surgical staging. Sensitivity, specificity, accuracy, and predictive values were then calculated based on pathology results. A retrospective review of the records was performed to determine how PET results affected clinical treatment decisions. Forty-seven of 50 patients had non-small-cell lung cancer. The prevalence of pathologically confirmed mediastinal and hilar involvement was 38%. The sensitivity, specificity, and accuracy of mediastinal disease staging were as follows: CT alone = 73%, 77%, 76%; PET alone = 73%, 94%, 87%; PET + CT = 82%, 96%, 91%, respectively. PET was more specific and accurate than CT (p = 0.025). The results of PET changed management decisions in 12 of 50 cases (24%). Using FDG-PET in conjunction with CT scanning provides the most accurate staging of mediastinal disease in lung cancer by contributing complementary information. Furthermore, PET can affect clinical decision-making and allow some patients considered unresectable a chance for resection. PMID- 10683078 TI - The roles of chemotherapy and surgery in gastric carcinoma and the influence of prognostic factors on survival. AB - In this study, we present the results of surgery and chemotherapy and the impact of various prognostic factors on survival in patients with gastric carcinoma with a follow-up of 6 years. All of the 328 cases were adenocarcinoma histologically and had a median age of 55 years. Median survival was 11 months, and the 5-year survival rate was 18%. Nonmetastatic cases were associated with improved survival as compared with the cases with metastatic disease (p<0.001). Patients with gastrectomy had improved survival (p<0.001). Subtotal gastrectomized patients had better survival rates in comparison to the total gastrectomized patients (p = 0.03). Addition of splenectomy to total gastrectomy and adjuvant chemotherapy did not influence survival rates (p>0.05). In metastatic patients, we determined beneficial effects of gastrectomy and chemotherapy on survival. The benefit was most predominant in chemoresponsive patients (p<0.001). Higher serum CA 19.9 levels in patients without metastases, higher serum lactate dehydrogenase and carcinoembryonic antigen levels in patients with metastases, and lower serum albumin levels in both stages were determined as significant predictors of poor survival. On multivariate analysis, only higher serum CA 19.9 level was the independent unfavorable prognostic factor of survival time in nonmetastatic patients (p = 0.008). In metastatic disease, older age (p = 0.03) and male gender (p = 0.05) were associated with poorer survival. In conclusion, gastric cancer is a great health problem, especially in developing countries, and we need more optimal approaches and treatment modalities for gastric cancer. PMID- 10683079 TI - Poorly differentiated carcinoma of the lung presenting with Lambert--Eaton myasthenic syndrome. AB - Lambert-Eaton myasthenic syndrome commonly seen in small-cell lung cancer represents an autoimmune reaction against antigens coexpressed by tumor and neurons. It is rarely seen with other histologic subtypes. Symptoms antedate the appearance of the neoplasm by weeks to years. Therapeutic options range from immunosuppression, plasmapheresis, pharmacologic facilitation of neuromuscular transmission, and definitive therapy of the primary tumor. This case report describes the rare association of Lambert-Eaton myasthenic syndrome with non small-cell lung cancer. PMID- 10683080 TI - A phase II trial of vinorelbine and cisplatin in previously untreated inoperable non--small-cell lung cancer. AB - Weekly vinorelbine injection with cisplatin had been used in treatment of non small-cell lung cancer. We performed a phase II trial to evaluate the efficacy and toxicity of a new schedule of vinorelbine and cisplatin in patients with previously untreated, inoperable (stage IIIB or stage IV) non-small-cell lung cancer. From April 1996 to May 1997, 52 patients were enrolled for study, and 50 patients were eligible and evaluable for both response and toxicity assessment. Therapy consisted of vinorelbine, 30 mg/m2, intravenously on days 1 and 5 of a 21 day cycle, and cisplatin 100 mg/m2 (reduced to 80 mg/m2 after the first seven patients) given on day 1. A total of 211 treatment courses were administered; the median number of cycles administered per patient was 4.5 (range: 1-6), the median dose intensity for vinorelbine was 16.9 mg/m2/week (84.4%), whereas that of cisplatin was 22.8 mg/m2/week (84.7%). Twenty-five patients responded to therapy for an overall response rate of 50%; one patient attained a complete response (2%). The main toxicities were vomiting, myelosuppression, and diarrhea, which included World Health Organization grade 3 or 4 nausea/vomiting (58% patients), anemia (41% patients), neutropenia (12% patients), and diarrhea (14%). The median duration of responses was 9 months. The median time to disease progression was 6.8 months (range 0.4-18.1 months). Median survival was 13 months, and 54% of patients were alive at 1 year. We conclude that this new schedule of vinorelbine and cisplatin achieves a high response with acceptable toxicity profile in patients with advanced non-small-cell lung cancer. PMID- 10683081 TI - Oral etoposide for Merkel cell carcinoma in patients previously treated with intravenous etoposide. AB - We describe three patients with advanced Merkel cell carcinoma who were treated with etoposide given orally for recurrent regional lymph node involvement 18 to 30 months after exposure to etoposide given intravenously. Etoposide given orally (100 mg/day) was given for 10 to 14 consecutive days and repeated every 21 to 28 days for a median of three courses (range: two to four). Toxicity was minimal and mainly hematologic. Two patients showed a complete response and one a partial response, all of very rapid onset. All three patients are alive 6, 9, and 42 months from the start of oral treatment. Two remain progression free, and one had a recurrence 1 month after completion of chemotherapy. We suggest that orally administered etoposide, a topoisomerase II inhibitor, has a strong antitumor effect in advanced Merkel cell carcinoma, even in patients previously treated parenterally with the same drug. This action may be explained by the greater dependence of the drug's efficacy on the duration of administration rather than the dose intensity. PMID- 10683082 TI - Phase II study of doxorubicin and paclitaxel as second-line chemotherapy of small cell lung cancer: a Hoosier Oncology Group Trial. AB - Forty-six evaluable patients with recurrent small-cell lung cancer were entered on a phase II Hoosier Oncology Group (HOG) protocol evaluating bolus doxorubicin 40 mg/m2 followed by paclitaxel 175 mg/m2 over 3 hours. Courses were repeated every 3 weeks for a maximum of 6 courses. Therapy was well-tolerated with grade III neurotoxicity in 5 patients (11%), grade III/IV emesis in 5 (11%), and grade III mucositis in 2 patients. One patient had grade IV myalgias and one patient had grade III cardiotoxicity. The main toxicity was myelosuppression. Twenty-nine patients (63%) had grade IV and 8 (17%) grade III granulocytopenia. Nine patients (20%) were hospitalized for granulocytopenic fever. There was no treatment related mortality. Nineteen of 46 patients (41%) had an objective response, including 3 complete remissions. Two of 14 patients with refractory disease (progression less than 3 months after initial therapy) responded, compared to 17 of 32 (52%) with sensitive disease (progression beyond 3 months of initial chemotherapy regimen). PMID- 10683083 TI - Warfarin is safe as secondary prophylaxis in patients with cancer and a previous episode of venous thrombosis. AB - The purpose of this study was to establish the safety and efficacy of sodium warfarin in the secondary prophylaxis of venous thrombosis in patients with cancer. This was an inception cohort study of patients enrolled in an anticoagulation clinic between July 1991 and October 1996. The rates of bleeding and recurrent thrombosis were evaluated in all the patients, and the results in patients with cancer (n = 104) were compared with those without cancer (n = 208). The rate of major hemorrhage was 0.4% and 0.3% per treatment month in the patients with cancer and those without cancer, respectively. The rates of recurrent thrombosis were 1.2% and 0.2% per treatment month in the patients with cancer compared with those without cancer, respectively. We conclude that warfarin is safe when used for the secondary prophylaxis of patients with cancer who have had a venous or arterial thrombosis, and the risk of major hemorrhage is not significantly different when compared with the risk in patients without cancer. The rate of recurrent thrombosis is approximately sixfold higher in patients with cancer being treated with warfarin for secondary prophylaxis of thrombosis compared with patients without cancer. Nonetheless, the rate of recurrent thrombosis is not overly excessive, and warfarin can be viewed as a relatively effective form of therapy for these patients. PMID- 10683084 TI - Thrombotic thrombocytopenic purpura in metastatic carcinoma of the breast. AB - We present a case of a 52-year-old woman with thrombotic thrombocytopenic purpura associated with progressive metastatic adenocarcinoma of the breast. The patient received plasma exchange therapy. Thrombocytopenic purpura resolved 2 months after discontinuation of plasma exchange while the patient received chemotherapy. After 3 more months, a fulminant relapse of the thrombocytopenic purpura developed, and there were signs of tumor progression. She died despite adequate treatment. We conclude that effective treatment of the underlying tumor can be crucial to control cancer-associated thrombocytopenic purpura. PMID- 10683085 TI - A phase I trial of gemcitabine and infusional 5-fluorouracil (5-FU) in patients with refractory solid tumors: Louisiana Oncology Associates protocol no. 1 (LOA 1). AB - The major purposes of this study were to determine the maximally tolerated dose (MTD), dose-limiting toxicity (DLT), toxicity profile, and antitumor activity of gemcitabine (GEM) (Gemzar) and 5-fluorouracil (5-FU) combination therapy when administered to patients with advanced solid tumors. GEM was administered intravenously over 30 minutes on days 1, 8, and 15, and 5-FU was administered as a continuous intravenous infusion from day 1 through day 15 of each 28-day treatment course. Seventeen patients (13 men and 4 women, median age 57, all previously treated with chemotherapy) were treated with 68 courses at 3 dose levels: 800/200, 1,000/200, and 1,000/300 [GEM (mg/m2/week)/ 5-FU (mg/m2/day)]. Two further patients were not fully evaluable for toxicity; one died from a probable pulmonary embolism, and one refused further treatment after developing grade II mucositis and dermatitis after her day 1 to 7 treatment. At the third dose level, 2 of 4 patients developed grade III mucositis; one also developed grade IV neutropenia with fever and grade III thrombocytopenia. Patient accrual then resumed at the second dose level. At this level, 10 patients were treated, with two developing grade III mucositis. One of these patients also developed grade IV dermatitis. No other patient developed grade III or IV side effects. Prophylactic dexamethasone was initiated after 4 of the first 7 patients (including 1 of the not fully evaluable patients) developed dermatitis-grade IV in 1 patient and grade II in the remaining 3 patients. After the steroids were initiated, 4 of the last 11 patients treated developed dermatitis, but grade 1 in all cases. One patient with metastatic gastric cancer achieved a near-complete response of his gastric mass and adrenal metastasis. Minor responses were achieved in a patient with colon carcinoma and a patient with an ethmoid sinus adenoid cystic carcinoma. The MTD and recommended dose for phase II clinical trials of GEM and 5-FU on the above schedule is 1,000 mg/m2 and 200 mg/m2 respectively, with mucositis as the DLT. PMID- 10683086 TI - Preliminary results of the use of Re-186-HEDP for palliation of pain in patients with metastatic bone disease. AB - We evaluated the effectiveness of Re-186-HEDP in 25 patients with painful metastatic bone disease. Twenty-five patients with known prostatic (n = 19), non small-cell lung cancer (n = 1) and breast cancer (n = 5) and multiple confirmed skeletal metastases were studied. All were taking analgesics daily (nonsteroidal antiinflammatory drugs/opiates). Re-186-HEDP (mean 35.2 mCi) was administered and patients were monitored for at least 50 days. In five patients, a repeat dose was administered 9 to 10 weeks later. The evaluation of the analgesic effect was based on a "pain diary" and by recording the use of analgesics. In 80% (20 of 25) of the patients, the effect was significant palliation, moderate in 3 patients (12%), and insignificant in 2 (8%). No significant myelotoxicity was observed. Transient pain flare was recorded in 8 of 25 patients. These results indicate that Re-186-HEDP can offer pain palliation in patients with painful bone metastases without being complicated by significant myelotoxicity. PMID- 10683087 TI - Short-course palliative radiotherapy in non-small-cell lung cancer: results of a prospective study. AB - From February 1993 to October 1997, 91 consecutive patients with inoperable (stage IIIB-IV) histologically confirmed non-small-cell lung cancer underwent palliative hypofractionated radiotherapy. Recently, the Medical Research Council studies on hypofractionated short-course radiotherapy (8.5 Gy x 2) have reported high control of symptoms caused by thoracic disease without toxicity. Based on these experiences and our previous positive trial on short-course radiotherapy (8 Gy x 2) in metastatic spinal cord compression, a prospective study of short course palliative radiotherapy in non-small-cell lung cancer was carried out. The regimen was 16 Gy given in two 8-Gy fractions, 1 week apart. Eighty-one patients were evaluable for response to treatment. Forty-eight (59%) patients were 65 years or older. Forty (49%) patients were naive to radiotherapy, whereas 41 (51%) had previous cisplatin-based chemotherapy. All but four stage IV patients (95%) had poor Eastern Cooperative Oncology Group performance status (i.e., 2-3). Clinical palliation was achieved in 62 (77%) patients. Performance status improved in 59 (73%) patients. The median palliation time ranged from 28% to 57% of patient survival. The median survival from the beginning of treatment was 148 days (range, 5-681 days). No difference in overall survival according to stage and previous chemotherapy was observed. Only performance status conditioned survival (performance status 1-2 vs. performance status 3; p = 0.0289). Short course radiotherapy gave good results in terms of clinical palliation for thoracic symptoms, even in patients with poor performance status and pretreated with chemotherapy. The median palliation time was approximately 50% of patient survival time. Treatment was generally well tolerated-only 4 (5%) patients experienced World Health Organization grade III dysphagia. No late toxicity was recorded. The two-fraction regimen had social and economic advantages compared with the conventional ones. PMID- 10683088 TI - Standard dose (Mayo regimen) 5-fluorouracil and low dose folinic acid: prohibitive toxicity? AB - Despite the perception that standard 5-fluorouracil/folinic acid (5-FU/FA) (425 mg/m2 per day and 20 mg/m2 per day intravenously once daily x 5 every 4 or 5 weeks) is well tolerated, we have been impressed by toxicity seen and frequent need for dose modification. We performed a retrospective analysis to quantitate the proportion of patients experiencing toxicity and attempted to identify associated clinical characteristics. One hundred thirty-four patients received 5 FU/FA at standard doses described by the Mayo regimen. Patient characteristics were as follows: female 35%, median age 66 years, Eastern Cooperative Oncology Group performance status less than or equal to 2, 96%. Sixty-eight percent received chemotherapy for metastatic disease. Forty-seven patients (35%+/-8%) experienced significant toxicity and were unable to receive the second cycle as scheduled: 76% required dose reduction, 11% discontinued therapy (including two toxic deaths), 11% discontinued therapy during the first cycle, and 2% required dose delay. Logistic regression was used to explore the following as predictors of toxicity: age, sex, performance status, adjuvant versus metastatic setting, prior chemotherapy, prior radiation, mean corpuscular volume, red blood cell distribution width, albumin, alkaline phosphatase, aspartate aminotransferase, bilirubin, and calculated creatinine clearance. No clinical characteristic was found to predict toxicity. Only high bilirubin approached statistical significance. We conclude that standard 5-FU/FA, when used in the general population, is associated with significant toxicity. Known clinical characteristics are not helpful in predicting toxicity. The lack of previous formal phase I evaluation of this regimen of 5-FU/FA raises concerns regarding its safety and generalizability in clinical practice. PMID- 10683089 TI - Synchronous primary cancers of the breast and cervix: planning multidisciplinary primary treatment [clinico-pathological conference]. AB - Multiple metachronous primary malignancies are becoming increasingly frequent; however, multiple synchronous primary malignancies are still unusual. We report the case of a 61-year-old woman with synchronous stage IIIB ductal carcinoma of the left breast and FIGO stage IB2 squamous cell carcinoma of the cervix. The patient was treated initially every 4 weeks with a 24-h intravenous infusion of paclitaxel (175 mg/m2) followed by a 1-h infusion of carboplatin (area under the curve of 5 mg/ml x min) with concurrent irradiation of the pelvis. Significant toxic reactions including nausea, vomiting, and diarrhea required hospitalization or outpatient intravenous fluids and antiemetics. After four cycles of chemotherapy, the breast cancer was in complete clinical remission, and the patient underwent a modified radical mastectomy with axillary lymph node dissection. Pathologic findings revealed a few microscopic foci of residual infiltrating ductal carcinoma exhibiting a marked treatment effect; none of the 14 axillary lymph nodes removed showed evidence of metastatic tumor. A near complete pathologic response of the breast cancer and a complete clinical response of the cervical cancer were obtained. Adjuvant chemotherapy for the breast cancer was then initiated, followed by radiation and hormonal therapy. PMID- 10683090 TI - Correlation between water proton spin lattice relaxation time and radiation tolerance dose in normal human tissue. PMID- 10683091 TI - Exercise and insulin sensitivity: a review. AB - Physical activity has a beneficial effect on insulin sensitivity in normal as well as insulin resistant populations. A distinction should be made between the acute effects of exercise and genuine training effects. Up to two hours after exercise, glucose uptake is in part elevated due to insulin independent mechanisms, probably involving a contraction-induced increase in the amount of GLUT4 associated with the plasma membrane and T-tubules. However, a single bout of exercise can increase insulin sensitivity for at least 16 h post exercise in healthy as well as NIDDM subjects. Recent studies have accordingly shown that acute exercise also enhances insulin stimulated GLUT4 translocation. Increases in muscle GLUT4 protein content contribute to this effect, and in addition it has been hypothesized that the depletion of muscle glycogen stores with exercise plays a role herein. Physical training potentiates the effect of exercise on insulin sensitivity through multiple adaptations in glucose transport and metabolism. In addition, training may elicit favourable changes in lipid metabolism and can bring about improvements in the regulation of hepatic glucose output, which is especially relevant to NIDDM. It is concluded that physical training can be considered to play an important, if not essential role in the treatment and prevention of insulin insensitivity. PMID- 10683092 TI - Dietary creatine monohydrate supplementation increases satellite cell mitotic activity during compensatory hypertrophy. AB - Nutritional status influences muscle growth and athletic performance, but little is known about the effect of nutritional supplements, such as creatine, on satellite cell mitotic activity. The purpose of this study was to examine the effect of oral creatine supplementation on muscle growth, compensatory hypertrophy, and satellite cell mitotic activity. Compensatory hypertrophy was induced in the rat plantaris muscle by removing the soleus and gastrocnemius muscles. Immediately following surgery, a group of six rats was provided with elevated levels of creatine monohydrate in their diet. Another group of six rats was maintained as a non-supplemented control group. Twelve days following surgery, all rats were implanted with mini-osmotic pumps containing the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) to label mitotically active satellite cells. Four weeks after the initial surgery the rats were killed, plantaris muscles were removed and weighed. Subsequently, BrdU-labeled and non-BrdU-labeled nuclei were identified on enzymatically isolated myofiber segments. Muscle mass and myofiber diameters were larger (P < 0.05) in the muscles that underwent compensatory hypertrophy compared to the control muscles, but there were no differences between muscles from creatine-supplemented and non-creatine supplemented rats. Similarly, compensatory hypertrophy resulted in an increased (P < 0.05) number of BrdU-labeled myofiber nuclei, but creatine supplementation in combination with compensatory hypertrophy resulted in a higher (P < 0.05) number of BrdU-labeled myofiber nuclei compared to compensatory hypertrophy without creatine supplementation. Thus, creatine supplementation in combination with an increased functional load results in increased satellite cell mitotic activity. PMID- 10683093 TI - Dehydroepiandrosterone (DHEA) rather than testosterone shows saliva androgen responses to exercise in elite female handball players. AB - The aim of the investigation was to evaluate in fourteen elite female handball players the resting values of saliva levels of testosterone -- T -- and dehydroepiandrosterone -- DHEA -- in comparison with ten sedentary women and the response to exercise of the same hormones in handball players. Saliva samples were taken from the handball players upon wakening (resting values) (8 a. m.), 5 minutes before and after a simulated handball match (6 p. m.; 8 p. m.) and upon wakening the following morning (8 a. m. + 24 h). The samples of saliva from the sedentary controls were taken at the same time of day but the controls did not perform any exercise. Resting hormonal levels were lower in handball players than in controls (P < 0.01). The exercise did not induce significant changes in T and DHEA salivary concentrations in handball players. Positive correlations between testosterone concentrations and DHEA were observed at all the time points studied. These findings showed that the T and DHEA concentrations followed a remarkably similar pattern. Considering the very low concentrations of testosterone in women, in particular in saliva, and its biosynthetic pathway, we suggest that measurements of DHEA could serve as a substitute for testosterone measurements to study training responses in elite sportswomen. PMID- 10683094 TI - Reduction of the plasma concentration of C-reactive protein following nine months of endurance training. AB - An intense physical exercise induces an inflammatory reaction as demonstrated by the delayed increase in blood of acute phase proteins and among them of C reactive protein (CRP). There is also evidence for a diminished acute phase reaction due to regular exercise suggesting a suppression of the inflammatory response through training. With this background CRP was measured by a sensitive enzyme immunoassay under resting conditions before and after 9 months of training in 14 subjects preparing for a marathon with the aim of studying the effect of training on the base-line CRP concentration. The mean distance run per week increased significantly from 31 +/- 9 km at the beginning to 53 +/- 15 km after 8 months of training (p < 0.01). The aerobic capacity rose significantly after training as demonstrated by the increase of running velocity during a maximal treadmill test from 3.82 +/- 0.29 m/s pre-training to 4.17 +/- 0.17 m/s post training at a blood lactate concentration of 4 mmol/L (p < 0.01). In 10 of 12 runners base-line CRP was diminished after training in spite of a continuous increase of training intensity. The CRP median fell from 1.19 mg/L before to 0.82 mg/L after training (p < 0.05). Since intense physical exercise is known to be associated with an inflammatory reaction of muscles and tendons, the CRP decrease was unexpected. In 2 subjects the CRP concentration rose markedly because of a borrelia infection and a knee injury, respectively. These values were caused by a pathological condition and were not considered for the statistical evaluation. In 10 non-training control subjects the CRP median did not change significantly during the same 9 months period. The decrease of the CRP base-line concentration after training suggests that intensive regular exercise has a systemic anti inflammatory effect. This is of particular interest with regard to several recent reports confering on the concentration of CRP in plasma a predictive value for the risk of cardiac infarction, venous thrombosis or stroke. PMID- 10683095 TI - The effect of free glutamine and peptide ingestion on the rate of muscle glycogen resynthesis in man. AB - The present study investigated previous claims that ingestion of glutamine and of protein-carbohydrate mixtures may increase the rate of glycogen resynthesis following intense exercise. Eight trained subjects were studied during 3 h of recovery while consuming one of four drinks in random order. Drinks were ingested in three 500 ml boluses, immediately after exercise and then after 1 and 2 h of recovery. Each bolus of the control drink contained 0.8 g x kg(-1) body weight of glucose. The other drinks contained the same amount of glucose and 0.3 g x kg(-1) body weight of 1) glutamine, 2) a wheat hydrolysate (26% glutamine) and 3) a whey hydrolysate (6.6% glutamine). Plasma glutamine, decreased by approximately 20% during recovery with ingestion of the control drink, no changes with ingestion of the protein hydrolysates drinks, and a 2-fold increase with ingestion of the free glutamine drinks. The rate of glycogen resynthesis was not significantly different in the four tests: 28 +/- 5, 26 +/- 6, 33 +/- 4, and 34 +/- 3 mmol glucosyl units x kg(-1) dry weight muscle x h(-1) for the control, glutamine, wheat- and whey hydrolysate ingestion, respectively. It is concluded that ingestion of a glutamine/carbohydrate mixture does not increase the rate of glycogen resynthesis in muscle. Glycogen resynthesis rates were higher, although not statistically significant, after ingestion of the drink containing the wheat (21 +/- 8%) and whey protein hydrolysate (20 +/- 6%) compared to ingestion of the control and free glutamine drinks, implying that further research is needed on the potential protein effect. PMID- 10683096 TI - The slow component of O2 uptake kinetics during high-intensity exercise in trained and untrained prepubertal children. AB - The aim of the present study was to investigate the O2 uptake slow component in prepubertal children of different aerobic capacity during high intensity exercise. Twenty-three (12 well-trained, T and 11 untrained, U subjects) 10-13 year old prepubertal children took part in 3 tests: one incremental test to determine the maximal aerobic power (PMA) and anaerobic threshold (LAT); two constant-power tests performed at intensities corresponding to 80%LAT and 90%PMA. Oxygen uptake (VO2), heart rate, ventilation (VE) and lactate ([L]s) were evaluated during each test. A monoexponential + linear term model (starting after phase 1) was used to assess VO2 kinetics during both constant-power tests. Our results showed that a slow component, represented by the linear coefficient (S) of the mathematical model, was present during the 90%PMA test only (S = 0.86 +/- 0.48 ml x min(-2) x kg(-1) for the whole population). No relationships were found between either S and VE or [L]s, showing that, at least in prepubertal children, these factors play a minor role in the explanation for the VO2 slow component. The slow component contributed approximately to the same amount of the total VO2 response in both groups (T: 21.4 +/- 8.0, U: 19.3 +/- 3.9%, ns). In conclusion, as previously described in adults, our data demonstrated the existence of a slow component in prepubertal children during high-intensity exercise. Moreover, this slow component was similar in trained and untrained children, exercising at the same relative intensity. PMID- 10683097 TI - The stability of lactate concentration in preserved blood microsamples. AB - This study aimed to determine the stability of lactate concentration in blood samples preserved and stored using methods practical for field testing and experimental applications. Whole blood microsamples were obtained from venous samples drawn from 10 healthy subjects following bouts of moderate (approximately 5 mmol x l(-1), n = 12), or intense (approximately 10 mmol x l(-1), n = 12), treadmill exercise. Samples were analysed fresh (2 x 25 microl), or placed in preservative-containing tubes (12 x 75 microl) and analysed directly, or after storage at room temperature (RT) or 4 degrees C, for 1 h, 18 h, 2 d, 3 d or 7 d, or at -20 degrees C for 7 d. In comparison to preserved samples assayed directly after collection, lactate levels in all RT samples had declined significantly, whereas the 4 degrees C samples had not changed, by 2 d post-collection. After 7 d of refrigeration, the absolute value of the difference from lactate levels in samples measured after collection (mean +/- SD) was 0.38 +/- 0.34 mmol x l(-1), or 5.3 +/- 4.3%; with freezing, this difference was 0.27 +/- 0.27 mmol x l(-1), or 3.6 +/- 3.0%. These differences were less than the daily variation in the analyser readings of a 10 mmol x l(-1) standard, indicating that the blood preservation and storage methods identified herein are suitable for use during exercise testing. PMID- 10683098 TI - Joint excursion, handle velocity, and applied force: a biomechanical analysis of ergonometric rowing. AB - Rowers may sacrifice on-water technique during ergonometric training in an attempt to increase stroke output. This cross-sectional study aimed to identify characteristics of ergonometric rowing technique that could be potentially detrimental to an effective and safe on-water performance. Joint excursion, handle velocity, and applied force were measured in 44 athletes while they performed a 2500 meter race on an instrumented ergometer. Results on four subjects are presented here. Their performance is compared to that of a Barcelona Olympic and World champion rower with 12 years of experience to illustrate how athletes deviate from standard on-water technique. Kinematic data showed knee joint oscillations and out-of-phase hip and knee joint reversals. Horizontal handle velocity curves indicated that higher stroke rates were achieved by a decrease in recovery time. Vertical handle velocity curves exhibited bi directional variations. The largest amplitude occurred at the end of the drive phase during an upward displacement of the handle that was associated with a jerk in the applied force. Force-time curves at different stroke rates showed greater variability in the initial portion of the drive phase. Perpetuation of these technique deviations may be detrimental to on-water performance. Biomechanical analyses may allow coaches to better monitor technique during ergonometric training. PMID- 10683099 TI - Heart rate and blood pressure variability during heavy training and overtraining in the female athlete. AB - We investigated heavy training- and overtraining-induced changes in heart rate and blood pressure variability during supine rest and in response to head-up tilt in female endurance athletes. Nine young female experimental athletes (ETG) increased their training volume at the intensity of 70-90% of maximal oxygen uptake (VO2max) by 125% and training volume at the intensity of < 70% of VO2max by 100% during 6-9 weeks. The corresponding increases in 6 female control athletes were 5% and 10%. The VO2max of the ETG and the control athletes did not change, but it decreased from 53.0 +/- 2.2 ml x kg(-1) x min(-1) to 50.2 +/- 2.3 ml x kg(-1) x min(-1) (mean+/-SEM, p < 0.01) in five overtrained experimental athletes. In the ETG, low-frequency power of R-R interval (RRI) variability during supine rest increased from 6 +/- 1 ms2 x 10(2) to 9 +/- 2 ms2 x 10(2) (p < 0.05). The 30/15 index (= RRI(max 30)/RRI(min 15), where RRI(max 30) denotes the longest RRI close to the 30th RRI and RRI(min 15) denotes the shortest RRI close to the 15th RRI after assuming upright position in the head-up tilt test), decreased as a result of training (analysis of variance, p = 0.05). In the ETG, changes in VO2max were related to the changes in total power of RRI variability during standing (r = 0.74, p < 0.05). Heart rate response to prolonged standing after head-up tilt was either accentuated or attenuated in the overtrained athletes as compared to the normal training state. We conclude that heavy training could increase cardiac sympathetic modulation during supine rest and attenuated biphasic baroreflex-mediated response appearing just after shifting to an upright position. Heavy-training-/overtraining-induced decrease in maximal aerobic power was related to decreased heart rate variability during standing. Physiological responses to overtraining were individual. PMID- 10683100 TI - A new index of coordination for the crawl: description and usefulness. AB - This study analyzes stroke phases and arm and leg coordination during front crawl swimming as a function of swim velocity and performance level. Forty-three swimmers constituted three groups based on performance level. All swam at three different swim velocities, corresponding to the paces appropriate for the 800 m, 100 m, and 50 m. The different stroke phases and the arm and leg coordination were identified by video analysis. Arm coordination was quantified using a new index of coordination (IdC), which expresses the three major modalities opposition, catch-up and superposition. Opposition, where one arm begins the pull phase when the other is finishing the push phase; catch up, which has a lag time (LT) between propulsive phases of the two arms; and superposition, which describes an overlap in the propulsive phases. The IdC is an index which characterizes coordination patterns by measure of LT between propulsive phases of each arm. The most important results showed that duration of the propulsive phases (B + C) increased significantly with increasing velocity: 43.1 +/- 3.3% for V800; 46.5 +/- 3% for V100 and 49 +/- 3% for V50. The arm and leg synchronization was modified in the sense of an increase in six-beat kick. The IdC increased significantly with velocity: IdCV800 = -7.6 +/- 6.4%; IdCV100 = 3.2 +/- 5.1% and IdCV50 = -0.9 +/- 5.6%. IdC increased also significantly with performance level: IdCG3 = -6.07 +/- 5.3%; IdCG2 = -3.9 +/- 4.2% and IdCG1 = 1.76 +/- 5.6% for the mean of the 3 velocity. The two extreme IdC were IdCG3V800 = -9.4 +/- 5.4% and IdCG1V50 = +2.53 +/- 4.4%. PMID- 10683101 TI - Energetically optimal cadence vs. freely-chosen cadence during cycling: effect of exercise duration. AB - The purpose of this study was to examine the relationship between cadence and oxygen consumption with exercise duration. Ten triathletes who trained regularly were examined. The first test was always a maximal test to determine maximal oxygen uptake (VO2max). The other sessions were composed of six submaximal tests representing 80% of the maximal power reached with VO2max (Pmax). During these tests submaximal rides with a duration of 30 min were performed. Each test represented, in a randomised order, one of the following pedal rates: 50, 65, 80, 95, 110 rpm and a freely-chosen rate. VO2, respiratory parameters, and heart rate were monitored continuously. Two periods, between the 3rd and the 6th minute and between the 25th and the 28th minute, were analysed. Results showed that when VO2 and heart rate were plotted against cadence, each curve could be best described by a parabolic function, whatever the period. Furthermore, a significant effect of period was found on energetically optimal cadence (70 +/- 4.5 vs. 86 +/- 6.2 rpm, P < 0.05). Only during the second period was no significant difference found between freely-chosen cadence (83 +/- 6.9 rpm) and energetically optimal cadence (P > 0.05). In conclusion, our results suggest that during prolonged exercise triathletes choose a cadence that is close to the energetically optimal cadence. A change of muscle fibre recruitment pattern with exercise duration and cadence would explain the shift in energetically optimal rate towards a higher pedal rate observed at the end of exercise. PMID- 10683102 TI - The effect of a sports drink on gastroesophageal reflux during a run-bike-run test. AB - The effects of different modes of prolonged exercise and different drinks on gastroesophageal reflux and reflux-related symptoms were examined. In a cross over design seven male triathletes performed two tests at one week intervals (50 min periods of alternately running, cycling and running at 70-75% VO2max), with supplementation of either a conventional sports drink (7% carbohydrates) or tap water. Gastroesophageal reflux (percentage time and number of periods esophageal pH < 4) was measured with an ambulant pH system before, during and after exercise. Percentage reflux time (+/- SEM) during running, cycling, running and recovery was 24.0 +/- 4.6, 8.2 +/- 4.8, 17.6 +/- 8.4 and 11.8 +/- 4.0 with carbohydrates and 7.4 +/- 2.9, 0 +/- 0, 2.4 +/- 1.4 and 0.2 +/- 0.2 with water, respectively. Reflux lasted longer during exercise as compared to the rest situation (5.6 + 1.4%), especially with carbohydrates, and lasted longer with carbohydrates than with water (P < 0.05; Wilcoxon signed rank test). In general, reflux lasted longer during running than during cycling (P < 0.05). Data on the number of reflux periods are concordant to these results. Chest pain was reported by one subject during running with carbohydrates. Heartburn during running was reported by two subjects with water and by one with carbohydrates. In conclusion, physical exercise increases gastroesophageal reflux, dependent on the mode of exercise and beverage used. PMID- 10683103 TI - Acute creatine supplementation in older men. AB - The hypothesis of this study was that short term creatine (Cr) ingestion in older individuals would increase body mass and exercise performance, as has been shown in younger subjects. Seventeen males 60-78 years old were randomly placed into two groups, Cr and placebo (P), and supplemented in double-blind fashion for 5 days. Subjects ingested either 5 g of Cr plus 1 g of sucrose 4x per day or 6 g of a sucrose placebo 4x per day. Isometric strength of the elbow flexors was assessed using a modified preacher bench attached to a strain gauge. Isokinetic exercise performance was assessed using an intermittent fatigue test of the knee extensors. Subjects performed 3 sets of 30 repetitions with 60 sec rest between sets. There was a small (0.5 kg) but statistically significant increase in body mass (p < 0.05) in the Cr group after supplementation. There was a significant overall interaction between groups in isokinetic performance from pre to post supplementation (group x time x set, p < 0.05). However, analysis of the groups separately revealed that the subjects in the Cr group demonstrated a small non significant increase in isokinetic performance while subjects in the P group demonstrated a small non-significant performance decrement. There was no significant difference in isometric strength between groups from pre to post supplementation. These data suggest that acute oral Cr supplementation does not increase isometric strength and only produces small increases in isokinetic performance and body mass in men over the age of 60. PMID- 10683104 TI - Incomplete restoration of immobilization induced softening of young beagle knee articular cartilage after 50-week remobilization. AB - The aim of this study was to characterize the biomechanical and structural changes in canine knee cartilage after an initial 11-week immobilization and subsequent remobilization period of 50 weeks. Cartilage from the immobilized and remobilized knee was compared with the tissue from age-matched control animals. Compressive stiffness, in the form of instant shear modulus (ISM) and equilibrium shear modulus (ESM) of articular cartilage, was investigated using an in situ indentation creep technique. The local variations in cartilage of glycosaminoglycan (GAG) concentration were measured with a microspectrophotometer after safranin O staining of histological sections. Using a computer-based quantitative polarized light microscopy method, collagen-related optical retardation, gamma, of cartilage zones were performed to investigate the collagen network of cartilage. Macroscopically, cartilage surfaces of the knee joint remained intact both after immobilization and remobilization periods. Immobilization caused significant softening of the lateral femoral and tibial cartilages, as expressed by ESM (up to 30%, p < 0.05). Remobilization restored the biomechanical properties of cartilage in the lateral condyle of tibia, but in the lateral condyle of femur ESM remained 15% below the control level (p = 0.05). The instant shear modulus was not changed either after immobilization or remobilization. The GAG content of the cartilage was slightly decreased after immobilization, especially in the superficial zone of cartilage, but the change was not statistically significant. After remobilization the intensity of safranin O content rose to control level. Neither immobilization nor remobilization had any effect on the gamma value of collagen fibril network either in the superficial or the deep zone at any of the test points. The changes of ESM were positively correlated with the alterations in GAG content of the superficial and deep zones after immobilization and remobilization. This confirms the key role of protoglycans in the regulation of the equilibrium stiffness of articular cartilage. As a conclusion, immobilization of the joint of a young individual may cause long-term, if not permanent, alterations of cartilage biomechanical properties. This may predispose joint to degenerative changes later in life. PMID- 10683105 TI - Avian behavioural neuroscience: past, present and future perspectives. AB - A survey of avian brain-behavior conference reports of the last 25 years reveals that neither the avian species studied nor the types of scientific questions asked have changed very much since the first such conference report in 1974. The birds studied tend, for the most part, to be pigeons, chickens, quail, and canaries. Because of the growing interest in avian vocalization and its neural control, one recent conference featured studies of canaries, zebra finches, and budgerigars. The topics of investigation at these conferences largely have involved sensory systems, mostly sensory and sensorimotor mechanisms with a heavy emphasis on vision and audition, as well as studies of learning and memory. Future research should expand the range of orders and species of birds studied so as to shed light on evolutionary trends within Aves as a whole. The scope of behavioral questions asked also should be broadened to include topics of neuroethological interest. PMID- 10683106 TI - Visual circuits of the avian telencephalon: evolutionary implications. AB - Birds and primates are vertebrates that possess the most advanced, efficient visual systems. Although lineages leading to these two classes were separated about 300 million years ago, there are striking similarities in their underlying neural mechanisms for visual processing. This paper discusses such similarities with special emphasis on the visual circuits in the avian telencephalon. These similarities include: (1) the existence of two parallel visual pathways and their distinct telencephalic targets, (2) anatomical and functional segregation within the visual pathways, (3) laminar organization of the telencephalic targets of the pathways (e.g. striate cortex in primates), and (4) possible interactions between multiple visual areas. Additional extensive analyses are necessary to determine whether these similarities are due to inheritance from a common ancestral stock or the consequences of convergent evolution based on adaptive response to similar selective pressures. Nevertheless, such a comparison is important to identify the general and specific principles of visual processing in amniotes (reptiles, birds, and mammals). Furthermore, these principles in turn will provide a critical foundation for understanding the evolution of the brain in amniotes. PMID- 10683107 TI - Functional subdivisions of the ascending visual pathways in the pigeon. AB - This study represents an attempt to examine an alternative view of the functional architecture of the ascending visual pathways in pigeons. According to this conception the pars dorsalis (GLd) of the thalamofugal system represents the lateral monocular field of view and is frontally blind to a large extent. The tectofugal system, on the other hand, processes frontal visual input within the framework of asymmetrical tectorotundal connections. As a result, the left, but not the right, rotundus should be able to integrate to an important degree the input from both eyes via the tecta of both hemispheres. Two lesion studies were conducted to test these assumptions. In the first psychophysical experiment, the visual acuity was determined in head-fixed pigeons. After thresholds were determined, stereotaxic lesions were placed in the GLd and/or the rotundus. Multiple regressions between structure specific lesion extents and postoperative threshold alterations demonstrated that only GLd lesions contributed to acuity reductions. In the second experiment the acuity threshold of pigeons under binocular and monocular conditions was determined in a conventional skinner box before GLd and/or rotundus lesions. Multiple regression analyses showed that rotundus--but not GLd lesions--contributed to performance losses. The left rotundus lesions were significantly related to threshold elevations under both monocular conditions, while the right rotundus only contributed together with the left rotundus to binocular performance. The double dissociation revealed in these experiments indicates that the ascending pathways in pigeons are functionally segregated and differentially process frontal and lateral as well as left- and right-sided inputs. PMID- 10683108 TI - Functional anatomy of the avian centrifugal visual system. AB - Although first described over a century ago, the centrifugal visual system (CVS) projecting to the retina still remains somewhat of an enigma with regard to its functional role in visually-guided behavior. The highly developed avian CVS has been the most extensively investigated and the anatomical organization of its two component centrifugal structures, the n. isthmo-opticus (NIO) and ectopic neurons (EN), including its afferent brainstem projections is reviewed. The results of double-labeling studies combining axonal tracing techniques and immunohistofluorescence have demonstrated GABA immunoreactivity (-ir) of interneurons within the neuropilar zone of the NIO, choline acetyltransferase (ChAT)-ir and nitric oxide synthase (NOS)-ir in the centrifugal cells of the NIO and EN as well as in the afferent projection neurons of layers 9/10 of the optic tectum. The data are discussed in terms of neurochemical and excitatory/inhibitory mechanisms within the different components of the avian CVS in relation to hypotheses which have implicated this system in visual attention and ground-feeding behavior. PMID- 10683109 TI - The dorsocaudal neostriatum of the domestic chick: a structure serving higher associative functions. AB - The dorsocaudal neostriatal (dNC) complex consists of at least three functionally distinct subregions and is part of an 'imprinting' pathway, which interconnects several forebrain regions that are known to be involved in juvenile learning. Based on its anatomical features, at least one subregion of the dNC complex, the neostriatum dorsocaudale (Ndc) may be considered as the equivalent of the mammalian polysensory association cortices. Several lines of evidence point to a role for this forebrain region in learning and memory formation. After auditory or visual imprinting changes of stimulus-evoked metabolic activities and of synaptic densities have been measured in the Ndc. Pharmacological behavioral studies revealed that the activation of NMDA receptors plays a critical role during this learning process and that NMDA receptor activation is required for the associated metabolic and synaptic changes. In addition to glutamatergic afferents, anatomical studies revealed a massive input from monoaminergic and peptidergic pathways into the dNC complex, suggesting a modulatory role for these systems during imprinting. The results presented here together with data from other avian species support the view that the dNc complex, and in particular the Ndc, plays an important role in juvenile and adult learning. PMID- 10683110 TI - The avian hippocampal formation: subdivisions and connectivity. AB - The avian hippocampal formation (HP) is considered to be homologous to the mammalian hippocampus on the basis of topography, developmental origin and its role in processing spatial memory. However, the morphological organization of the avian HP is very different from that of mammals and components similar to the subdivisions of the mammalian structure are not readily recognizable. In passerine birds, three spatially and morphologically distinct populations of Calbindin immunoreactive neurones are found in the dorsolateral (DL), dorsomedial (DM) and ventral (V) aspects of HP. Iontophoresis of Phaseolus vulgaris leucoagglutinin revealed three consistently different projection patterns arising from the different subregions. Generally, there is a medial-to-lateral topographical organization of efferents in relation to the septal complex. The DL region could be paralleled to the subiculum of mammals with its main projections to the basal ganglia, the limbic archistriatum, the lateral septum and the paraxial meso-diencephalic centres. The 'V' subdivision is likely to be homologous to the Ammon's horn of mammals with its commissural projections to the contralateral HP. Based on its purely intrinsic connectivity, the DM region could be a good candidate for an equivalent of the dentate gyrus. Nitric oxide synthase (NOS) containing neural structures display a specific distribution within the hippocampal subregions which is uniform in all passerine species studied. However, there is a marked difference in the level of diffuse neuropil reactivity between food-storers versus non-storers. Unlike the mammalian homologue, avian hippocampal NOS positive neurones do not show a near complete co-localization with the inhibitory transmitter GABA. PMID- 10683111 TI - Striato-telencephalic and striato-tegmental circuits: relevance to learning in domestic chicks. AB - Memory formation for a passive avoidance task in the domestic chick is likely to involve a hyperstriatum ventrale (IMHV)-archistriatum-lobus parolfactorius (LPO) arc. The present study summarises previous findings, relevant to this neural system, and is also supplemented with some recent data from our laboratory. Projections from the IMHV on the archistriatum, as well as from the archistriatum on the LPO, have been characterised using a combination of anterograde pathway tracing (Phaseolus lectin), and post-embedding GABA and glutamate immunocytochemistry. The majority of IMHV efferents have been found to synapse with dendritic spine heads and necks of densely spiny projection neurons of the ventral archistriatum, and the ultrastructure of synapses suggested a potent excitatory input. Similar synaptic connections of the excitatory type were ultrastructurally verified between ventral archistriatal afferent terminals and dendrites or spines of the LPO, suggesting an involvement of the medium sized spiny neurons, which are typical of the striatum. Although some of the IMHV boutons terminating in the archistriatum were immunoreactive to glutamate, this was not observed in the archistriatal-LPO pathway. Tegmental connections of the basal ganglia, in particular LPO, are also likely to play a role in processing of the avoidance response. We have demonstrated reciprocal connections between the LPO and dopaminergic (TH-positive) neurons of the substantia nigra and ventral tegmentum. Dopamine D1 receptors were upregulated bilaterally in the LPO following avoidance learning and this response was not accompanied by significant changes in the level of dopamine or its metabolites (HVA, DOPAC), as revealed by HPLC chromatography of brain samples dissected from the LPO of control and trained chicks. The dopamine receptor-related phosphoprotein DARPP-32 was localised in dendritic elements of the LPO, often forming asymmetric synapses with glutamate immunoreactive axon terminals. The findings are consistent with a scenario in which the striatum acts as a suppressor of natural pecking behaviour. Learned visual association with the target (bead) occurs in the IMHV and is relayed to the basal ganglia via the limbic archistriatum (amygdala equivalent), the latter introducing a motivational element (aversion, fear). Suppression of a brainstem pecking centre is likely to involve activation of the nigrostriatal (tegmentostriatal) dopaminergic circuit. PMID- 10683112 TI - Cellular correlates of stages of memory formation in the chick following passive avoidance training. AB - The process of memory formation has been investigated using the model of one trial passive avoidance training in the one-day old domestic chick. We have unraveled a biochemically coherent cascade of processes which, beginning with transient ion and neurotransmitter flux, and by way of a sequence of interacting pre- and post-synaptic intracellular signalling steps, results in gene activation and the synthesis of cell adhesion molecules which appear to be the effective agents in the structural processes involved in remodelling of synaptic and neuronal circuits. Further, in a related series of experiments we have shown that these biochemical and morphological changes are accompanied by significant changes in the neurophysiological status of the neurons on the IMHV and LPO, in particular in terms of their engagement in bouts of high-frequency firing. However, much remains to be clarified, particularly the meaning of the time dependent shifts in the location of the trace, and the ways in which these molecular and cellular events translate into changes in behavior in the animal. PMID- 10683113 TI - Early learning and the development of filial preferences in the chick. AB - Newly hatched domestic chicks (Gallus gallus domesticus) rapidly form a social preference for a conspicuous stimulus to which they are exposed. The learning process involved is known as filial imprinting. When chicks are exposed to an audio-visual compound stimulus, both auditory and visual learning are enhanced. The enhancement of visual imprinting is virtually abolished when chicks are exposed separately to the auditory element, either before or after training with the audio-visual compound. Simultaneous exposure to the two elements of the compound is superior to sequential exposure in achieving the enhancement of visual learning. These results are unlike Pavlovian conditioning, but are consistent with an interpretation of imprinting as a form of within-event learning, where links are formed between the representations of the elements of the compound, that can be weakened by separate exposure to an element. Apart from imprinting, chicks may show a developing predisposition to approach stimuli resembling conspecifics. The predisposition emerges in dark-reared chicks given some non-specific experience during a sensitive period, and is expressed as a relatively general preference for stimuli with a head and neck region. In the natural situation, the animal's response may be biased by the predisposition, and through imprinting it then learns the characteristics of individuals. PMID- 10683114 TI - The recognition memory of imprinting: biochemistry and electrophysiology. AB - A restricted part of the intermediate and medial part of the hyperstriatum ventrale (IMHV) of the domestic chick forebrain is pivotal to the learning process of imprinting and is probably the site at which information about an imprinting stimulus is stored. A range of learning-related changes occur in the IMHV between 1 and 24 h after training. The earliest change described is in Fos like immunoreactivity. There follow changes in phosphorylation of the protein kinase C substrate MARCKS, morphological changes in axospinous synapses, an increase in NMDA receptor number and increases in amounts of the major isoforms of the neural cell adhesion molecule and clathrin heavy chain. All but the change in Fos-immunopositivity occurs in the left, but not the right, IMHV. Insufficient nitric oxide synthase is available in the IMHV to support the hypothesis that nitric oxide is a retrograde messenger contributing to the effect on Fos-like immunoreactivity. In chicks anaesthetised approximately 24 h after imprinting training, the spontaneous mean neuronal firing rate is related to a preference score (a measure of learning). In unanaesthetised chicks 24 h after training, the responsiveness of some IMHV neurons is biassed specifically towards the imprinting stimulus.The responses of other neurons in the IMHV generalise across some features of the training stimulus, such as form or colour. Some neurons in the IMHV of unanaesthetised chicks are responsive to the distance of an imprinting stimulus from the chick; distance-sensitive neurons can be distinguished from distance-insensitive neurones by the action potential shape. PMID- 10683115 TI - Electrophysiological correlates of past history: in vitro studies of the IMHV of the domestic chick. PMID- 10683116 TI - Behavioural and neurophysiological aspects of sexual imprinting in zebra finches. AB - Sexual imprinting has been defined as the process by which young animals learn the characteristics of their future sexual partners. It is a two stage process including an acquisition period where features of the social environment are learnt, and a stabilization process by which, under the guidance of the previously acquired social information, a preference for a sexual partner is established and stabilized, so that it cannot be altered again subsequently. The stabilization process is short (1 h) and can be controlled experimentally. This allows for the design of experiments to examine the physiological events accompanying the imprinting process. During the stabilization process, four areas of the forebrain are more activated than in any other behavioural context. These are the hyperstriatum accessorium/dorsale (HAD), the archi-neostriatum caudale (ANC), the medial neo/hyperstriatum (MNH) and the lateral neo/hyperstriatum (LNH). Isolation during development reduces the spine density of neurons in HAD and ANC and enhances it in MNH and LNH. Subsequent exposure to a female (which stabilizes the previously acquired preference in behavioural experiments) for 1 week leads to an enhancement of spine densities in HAD and ANC, and to a reduction in MNH and LNH. The enhancement in HAD and ANC is reversible by a second isolation period after the exposure to a female, the reduction within MNH and LNH is not. This irreversibility indicates that the reduction process within MNH and LNH may be the anatomical manifestation of the imprinting process. The examination of spine densities in the four brain areas after two experiments which have been shown previously to affect the stabilization process in behavioural experiments, confirms this idea. PMID- 10683117 TI - Light experience and lateralization of the two visual pathways in the chick. AB - Using retrograde labelling with the fluorescent tracer rhodamine B isocyanate (RITC), we have examined the organisation of the thalamofugal and tectofugal visual projections to the forebrain of the young chick. In addition, we have investigated the influence of light exposure prior to hatching on the development of the tectofugal visual projections. Our results for the thalamofugal projections confirm those found previously; viz., that there are more projections from the left side of the thalamus to the right hyperstriatum of the forebrain than from the right side of the thalamus to the left hyperstriatum in males and females. The organisation of the tectofugal visual projections to the rotundal nuclei was more symmetrical (males only examined) although there was a trend towards a greater number of projections from the left optic tectum to its ipsilateral nucleus rotundus than from the right optic tectum to its ipsilateral nucleus rotundus. There are numerous projections from the optic tecta to their contralateral rotundal nuclei but, in contrast to reports for the pigeon, no marked asymmetry was present in these. The ratio of contralateral to ipsilateral projections revealed significant asymmetry for projections from the ventral regions of the optic tecta and symmetry from the dorsal regions. Thus both visual pathways of the chick have asymmetrical organisation but the asymmetry is much greater in the thalamofugal pathway. The slight asymmetry in the tectofugal projections may be determined by exposing the embryo to light just before hatching, as known to be the case for thalamofugal projections. PMID- 10683118 TI - The differential roles of right and left sides of the brain in memory formation. AB - A series of transitions in chick memory formation, with sharp standard timings, which were revealed by amnestic agents, coincide with a series of brief windows of enhanced recall ('retrieval events'), repeating with periods of 16 min (left hemisphere) and 25 min (right). Their timings were recently confirmed by: (1) the demonstration of a brief dip in recall 5 min after each left hemisphere event in the period 0-100 min, (2) spontaneous use of the eye providing direct input to the hemisphere undergoing an event, and (3) good performance in a delayed match to-sample task after 100 min, only at times of near, or exact coincidence (400 min) with right and left events. The exact coincidence is accompanied by a late episode of consolidation, as is a transition in memory formation in the zebra finch with precisely the same timing. Linkages between right and left versions of a single experience appear to be established as a result of near coincidences of events. The greater involvement of the left hemisphere in initiating such interactions appears to be responsible for a wide range of asymmetries described for interocular transfer and unilateral insult. PMID- 10683119 TI - Scientific and trade groups clash over 'authoritative' statements for health claims. PMID- 10683120 TI - Rosiglitazone approved for treatment of type 2 diabetes. PMID- 10683121 TI - Second selective COX-2 inhibitor receives marketing approval. PMID- 10683122 TI - Drug review and postmarketing surveillance programs are sound, but systems approach to risk management is needed, says FDA. PMID- 10683123 TI - Report examines strategies for reducing breast cancer risk. PMID- 10683124 TI - Translating an idea into a product at ASHP. PMID- 10683125 TI - Improving the care of hemophiliacs by managing the drug benefit. PMID- 10683126 TI - Improving ambulatory care through therapeutic interchange. PMID- 10683127 TI - Charting the demand for pharmacists in the managed care era. AB - The demand for pharmacists in the changing health care market is discussed. The Pharmacy Manpower Project (PMP) evolved out of concerns raised in the late 1980s, when the demand for pharmacists exceeded the supply. PMP collects, analyzes, and disseminates data on pharmacy work force variables. PMP's Subcommittee to Study Demand Issues was formed after the publication in 1995 of the Pew Health Professions Commission report projecting dramatic surpluses of pharmacists. In 1996-97, the PMP subcommittee held a series of sessions to discuss the future demand for pharmacists and their services. The panel identified a wide range of work force projections, but it concluded that medication management problems in the context of increasing prescription numbers and the emergence of data-driven health care support a scenario of a steadily increasing demand for pharmacists and pharmaceutical services. The data did not show that higher penetration by managed care is associated with smaller pharmacy staffs or job loss in institutions. There is little reason to expect the dramatic downsizing of the pharmacy work force predicted by the third report of the Pew Commission. However, retaining pharmacy roles that are useful to the system and satisfying to pharmacists will require a continuation of current proactive measures by the profession. PMID- 10683128 TI - Survival differences associated with treatment of cytomegalovirus retinitis in Maryland patients with AIDS, 1987-1994. AB - Differences in survival related to treatment of cytomegalovirus (CMV) retinitis in AIDS patients were studied. The medical records of adult AIDS patients who had been diagnosed with CMV retinitis in a Maryland inpatient facility between September 1987 and September 1994 were reviewed to assess determinants of survival, including treatment with ganciclovir and foscarnet, use of zidovudine, and demographic characteristics. The review was based on inpatient and outpatient medical records and computerized data from the Maryland HIV Information System. Of 212 AIDS patients with CMV retinitis, 123 (58.0%) were treated exclusively with ganciclovir, 55 (25.9%) received foscarnet only, and the remaining 34 (16.1%) received both ganciclovir and foscarnet at some point after their diagnosis for CMV retinitis. Patients who received both drugs survived significantly longer after the diagnosis than patients who received either drug by itself. The median time from diagnosis of CMV retinitis to death was 464 days for patients receiving both drugs, 225 days for ganciclovir recipients, and 202 days for foscarnet recipients. Other positive predictors of survival were male sex and use of zidovudine. Among Maryland adults with AIDS who were treated for CMV retinitis between September 1987 and September 1994, the most common treatment for the eye infection was ganciclovir. Patients receiving both ganciclovir and foscarnet survived longer than those treated with either drug alone. PMID- 10683129 TI - Impact of interruptions and distractions on dispensing errors in an ambulatory care pharmacy. AB - A possible association between interruptions and distractions and the occurrence of dispensing errors was investigated. Fourteen pharmacists and 10 technicians in an ambulatory care pharmacy at a general medical-surgical hospital were tested for distractibility by using the group embedded figures test (GEFT) as well as for visual acuity and hearing. They were videotaped as they filled prescriptions during a 23-day period in 1992. A study investigator compared each filled prescription with the physician's written order, noted details of deviations, verified with the pharmacist any errors that occurred, and asked the pharmacist to correct the error if necessary. Interruptions and distractions were detected and characterized by reviewing the videotapes. None of the study participants had significant hearing or visual impairment. There was a significant association between GEFT scores and error rates. A total of 5072 prescriptions were analyzed, and 164 errors were detected, for an overall error rate of 3.23%. Wrong label information was the most common type of error (80% of errors detected). A total of 2022 interruptions (mean +/- S.D. per half hour per subject, 2.99 +/- 2.70) and 2457 distractions (mean +/- S.D. per half hour per subject, 3.80 +/-3.17) were detected. The error rate for sets of prescriptions with one or more interruptions was 6.65% and for sets during which there were one or more distractions, 6.55%. Interruptions and distractions per half hour were both significantly associated with errors. In an ambulatory care pharmacy, interruptions and distractions over a half-hour period were associated with dispensing errors, a majority of which involved incorrect label information. PMID- 10683130 TI - Creating a computerized database from administrative claims data. AB - The creation of a computerized database from Medicaid administrative claims data for research purposes is described. Researchers should consult with computer experts at their institution before selecting software for data manipulation and conversion. It is essential to have an accurate layout of the file record before attempting to convert raw claims data into data sets or other data formats. The location of data elements within the claim will vary depending on whether the record comes from a provider, an institution, or a pharmacy. Each claim contains a common header, a variable header, and a claim detail section. The difficulty in analyzing data elements within a claim detail lies in locating the starting point of the claim detail section. So that data elements not in character or numeric formats can be converted, the file record layout must describe the exact format of each data element and its COBOL notation. A data element dictionary is necessary for translating data element coding into usable data. Data elements not necessary for any planned analysis must be eliminated. The data are then "cleaned" to remove any denied or reversed claims and claims that contain incomplete or erroneous data. Regardless of the format data are obtained in, an accurate file record layout and a data element dictionary are essential to the conversion of administrative claims data into a computerized database for data analysis and research purposes. PMID- 10683131 TI - Economic and clinical impact of a pharmacy-based filgrastim protocol in oncology patients. AB - The effects of a clinical protocol for filgrastim use in oncology patients were studied. A chart review was conducted for every fourth oncology inpatient who received filgrastim at a community hospital between January and June 1996 to determine how filgrastim was being used in the hospital's oncology patients. The results were presented to the oncology committee, and a filgrastim protocol was implemented. The protocol stated that filgrastim would be discontinued when the absolute neutrophil count (ANC) was > or =1500 cells/mm3 for two days after the neutrophil nadir. Six months after the protocol was implemented, a follow-up evaluation was conducted by reviewing the chart for every fourth oncology patient who received filgrastim between November 1996 and April 1997. Twenty-one patient charts were reviewed before the protocol was implemented, and 34 charts were reviewed after implementation. The results showed there was compliance with the protocol for 19 (76%) of the 25 patients evaluable for compliance. Sixty-seven percent of patients were febrile before the protocol was implemented, and 56% were febrile afterward. Ten percent of patients had documented infections before implementation, compared with 12% afterward. The average ANC at which filgrastim was discontinued before and after the protocol was implemented was 6839 and 5538 cells/mm3, respectively. Filgrastim was discontinued by a pharmacist in 32% of cases. A $22,416 cost saving was achieved in the first six months after protocol implementation, with no compromise in clinical efficacy. A pharmacy-based protocol for discontinuing filgrastim therapy in oncology patients saved a community hospital more than $22,000 in the first six months withno adverse impact on the drug's effectiveness. PMID- 10683132 TI - Prevalence and treatment of heart failure in elderly long-term-care patients. PMID- 10683133 TI - Impact of pharmacist interventions on hospital readmissions for heart failure. PMID- 10683134 TI - Opportunities for pharmacy specialists as the delivery of health care changes. PMID- 10683135 TI - Developing a care plan. PMID- 10683136 TI - Clinical usefulness of urokinase after dilution and freezing. PMID- 10683137 TI - Doxazosin-to-terazosin switch for benign prostatic hyperplasia. PMID- 10683138 TI - Electronic prescriptions in pharmacy. PMID- 10683139 TI - Bad medical writing as a barrier to communication. PMID- 10683140 TI - Cross-regulation of the Wnt signalling pathway: a role of MAP kinases. AB - The Wnt signal transduction pathway regulates various aspects of embryonal development and is involved in cancer formation. Wnts induce the stabilisation of cytosolic (beta)-catenin, which then associates with TCF transcription factors to regulate expression of Wnt-target genes. At various levels the Wnt pathway is subject to cross-regulation by other components. Recent evidence suggests that a specific MAP kinase pathway involving the MAP kinase kinase kinase TAK1 and the MAP kinase NLK counteract Wnt signalling. In particular, homologues of TAK1 and NLK, MOM-4 and LIT-1, negatively regulate Wnt-controlled cell fate decision in the early Caenorhabditis elegans embryo. Moreover, TAK1 activates NLK, which phosphorylates TCFs bound to (beta)-catenin. This blocks nuclear localization and DNA binding of TCFs. Since TAK1 is activated by TGF-(beta) and various cytokines, it might provide an entry point for regulation of the Wnt system by other pathways. In addition, alterations in TAK1-NLK might play a role in cancer. PMID- 10683141 TI - The parting of the endothelium: miracle, or simply a junctional affair? AB - Leukocyte extravasation from the blood across the endothelium is vital for the functioning of the immune system. Our understanding of the early steps of this process has developed rapidly. However, it is still unclear how leukocytes undergo the final step, migrating through the junctions that mediate adhesion between adjacent endothelial cells, while preserving the barrier function of the endothelium. The first stage of transmigration - tethering and rolling - is mediated by interactions between selectins on the surface of leukocytes and glycosylated proteins such as GlyCAM-1 on the surface of endothelial cells. Stimulation of the leukocyte by chemokines then induces tight adhesion, which involves binding of activated leukocyte integrins to endothelial ICAM-1/VCAM-1 molecules. Passage of the leukocyte across the endothelium appears to require delocalization of certain endothelial cell molecules and proteolytic degradation of junctional complexes. PMID- 10683142 TI - Ras-independent oncogenic transformation by an EGF-receptor mutant. AB - Mutations in the ligand-binding domain of the epidermal growth factor receptor have been identified in several types of human cancers, including malignant gliomas. These mutations render signaling by this receptor to be constitutively ligand-independent. In fibroblasts transformed with ligand-independent epidermal growth factor receptor mutants, there is a correlation between the formation of a unique phosphotyrosine protein complex and oncogenic transformation. This phosphoprotein complex includes Grb2, Shc, Sos, tyrosine-phosphorylated form of caldesmon, and two, as yet, unidentified proteins. The presence of Grb2, Shc, and Sos in this complex implicates Ras in ligand-independent signaling by these oncogenic epidermal growth factor receptor mutants. We, therefore, have used retroviral co-infections of cultured primary fibroblasts to determine if Ras activation is required for phosphoprotein complex formation, stress fiber loss, or transformation. As predicted, expression of a dominant-negative Ras mutant (N17Ras) completely abrogates ligand-stimulated soft agar colony growth of primary fibroblasts. In contrast, N17Ras expression has no effect on v-ErbB mediated stress fiber disassembly, soft agar colony growth, or phosphoprotein complex assembly. In addition, our data suggest that ligand-dependent Ras activation may be suppressed by oncogenic v-ErbB expression. Together these observations suggest that oncogenic signaling by v-ErbB does not require Ras activation, and implicate an alternative signal transduction pathway in ligand independent epidermal growth factor receptor oncogenic signaling. PMID- 10683143 TI - Reconstitution of microtubule nucleation potential in centrosomes isolated from Spisula solidissima oocytes. AB - Treatment of isolated Spisula solidissima centrosomes with KI removes (gamma) tubulin, 25 nm rings, and their microtubule nucleation potential, revealing the presence of a filamentous lattice, the 'centromatrix'. Treatment of this centromatrix with Spisula oocyte extract results in the binding of (gamma) tubulin and 25 nm rings, and the recovery of microtubule nucleation potential. Fractionation of this extract resulted in the separation of elements that are required for the recovery of microtubule nucleation potential. We show that some, but not all, of the elements needed cosediment with microtubules. Further, extracts prepared from activated (meiotic) and non-activated (interphase) Spisula oocytes, CHO cells blocked in S phase, Drosophila embryos and Xenopus oocytes all support the recovery of microtubule nucleation potential by the Spisula centromatrix. These results demonstrate that components necessary for centrosome dependent microtubule nucleation are functionally conserved and abundant in both interphase and meiotic/mitotic cytoplasm. PMID- 10683144 TI - DNA-binding activity of the N-terminal cleavage product of poly(ADP-ribose) polymerase is required for UV mediated apoptosis. AB - The role of the N-terminal cleavage product of poly(ADP-ribose) polymerase (PARP) on UV mediated apoptosis was investigated in cultured HeLa cells. Ultrastructural analysis of cells expressing caspase-resistant PARP (PARP(D214A)) revealed the typical features of necrosis following UV treatment. However, cells co-expressing PARP(D214A) with the N-terminal fragment of PARP containing the DNA-binding domain underwent apoptosis instead of necrosis. In this study, we have demonstrated that the DNA-binding activity of the N-terminal fragment of PARP is important for the execution of apoptosis. Point mutations were introduced in the DNA-binding sites of the N-terminal fragment. Cells co-expressing PARP(D214A) with the mutated N-terminal fragments neither stimulated apoptosis nor prevented necrosis in response to UV irradiation. The present study proposes that the DNA binding activity of the N-terminal fragment of PARP in UV treated cells prevents cellular ATP depletion, a mechanism by which necrotic cell death is triggered. PMID- 10683145 TI - Formation of hemidesmosome-like structures in the absence of ligand binding by the (alpha)6(beta)4 integrin requires binding of HD1/plectin to the cytoplasmic domain of the (beta)4 integrin subunit. AB - Hemidesmosomes are adhesion structures that mediate anchorage of epithelial cells to the underlying basement membrane. We have previously shown that the (alpha)6(beta)4 integrin can induce the assembly of these multi-protein structures independent of binding to its ligand laminin-5 (ligand-independent formation of hemidesmosomes). Our results suggested a role for HD1/plectin, which binds to the cytoplasmic domain of the (beta)4 integrin subunit, in controlling the clustering of hemidesmosomal components at the basal side of the cell. Using keratinocytes derived from patients lacking HD1/plectin, we now show that ligand independent formation of hemidesmosomal clusters indeed requires HD1/plectin, in contrast to the ligand-dependent assembly of hemidesmosomes. No clustering of the (alpha)6(beta)4 integrin, or of the bullous pemphigoid antigens BP180 and BP230, was seen when HD1/plectin-deficient keratinocytes were plated on fibronectin or type IV collagen. In (&bgr;)4-deficient keratinocytes, expression of an interleukin 2 receptor (IL2R) transmembrane chimera containing the (beta)4 cytoplasmic tail with the mutation R1281W, which abrogates HD1/plectin binding, resulted in a diffuse distribution of the chimeric receptor. In contrast, a (beta)4(R1281W) mutant that can associate with (alpha)6 and bind ligand, was found to be directed to the basal surface of the cells, at sites where laminin-5 was deposited. In addition, this mutant induced clustering of BP180 and BP230 at these sites. Together, these results show that the formation of hemidesmosomes requires binding of either ligand or HD1/plectin to the (beta)4 integrin subunit. Intriguingly, we found that IL2R/(beta)4 chimeras become localized in pre existing hemidesmosomes of HD1/plectin-deficient keratinocytes, and that this localization requires a domain in the (beta)4 cytoplasmic tail that is also required for HD1/plectin binding (residues 1115-1356). Because this part of (beta)4 lacks the BP180 binding site, and since we show in this study that it is unable to interact with the same part on another (beta)4 molecule, we suggest that the chimera becomes incorporated into hemidesmosomes of HD1/plectin deficient keratinocytes by interacting with an as yet unidentified hemidesmosomal component. PMID- 10683146 TI - p39 activates cdk5 in neurons, and is associated with the actin cytoskeleton. AB - Cyclin-dependent kinase 5 (cdk5) is a small serine/threonine kinase that displays close sequence homology to the mitotically active cyclin-dependent kinases. Cdk5 has been shown to play an essential role in the development of the nervous system, including neuronal migration and neurite outgrowth. Cdk5 activation requires the presence of a regulatory activator such as p35. cdk5 -/- mice have much more extensive defects in the development of the nervous system than p35 -/- mice, leading to the speculation that other regulatory activators of cdk5 exist. Indeed, p39 is a p35 related protein isolated by sequence homology to p35. We show here that p39 associates with cdk5 in brain lysates, and that this complex is active in phosphorylation of histone H1. By extensive characterization of p39 subcellular localization in different cell types, we demonstrate the presence of p39 in lamellipodial and fillopodial structures of cells and in growth cones of neurons. We show that p39 colocalizes with actin, and cofractionates with the detergent insoluble cytoskeleton from brain. Further, p39 coimmunoprecipitates with actin in brain lysates. Finally, disruption of the actin cytoskeleton alters p39 subcellular localization as well as kinase activity of the p39/cdk5 complex. Therefore, our results reveal the existence of the p39/cdk5 complex in vivo and suggest that it might play a role in regulating actin cytoskeletal dynamics in cells. PMID- 10683147 TI - Inhibiting cadherin function by dominant mutant E-cadherin expression increases the extent of tight junction assembly. AB - Previous studies have shown that induction of cadherin-mediated cell-cell adhesion leads to tight junction formation, and that blocking cadherin-mediated cell-cell adhesion inhibits tight junction assembly. Here we report analysis of tight junction assembly in MDCK cells overexpressing a mutant E-cadherin protein that lacks an adhesive extracellular domain (T151 cells). Mutant E-cadherin overexpression caused a dramatic reduction in endogenous cadherin levels. Despite this, tight junction assembly was extensive. The number of tight junction strands observed by freeze-fracture electron microscopy significantly increased in T151 cells compared to that in control cells. Our data indicate that the hierarchical regulation of junctional complex assembly is not absolute, and that inhibition of cadherin function has both positive and negative effects on tight junction assembly. PMID- 10683148 TI - Preferential association of syntaxin 8 with the early endosome. AB - Members of the syntaxin family play a fundamental role in vesicle docking and fusion of diverse transport events. We have molecularly characterized syntaxin 8, a novel member of the syntaxin family. The nucleotide sequence of cloned rat cDNA predicts a polypeptide of 236 residues with a carboxyl-terminal 18-residue hydrophobic domain that may function as a membrane anchor. Characteristic of syntaxins, syntaxin 8 also contain regions that have the potential to form coiled coil structures. Among the known syntaxins, syntaxin 8 is most homologous to syntaxin 6 which is predominantly associated with the trans-Golgi network (TGN). The syntaxin 8 transcript is detected in all rat tissues examined by northern blot. Antibodies against recombinant syntaxin 8 recognize a 27 kDa protein that is enriched in membrane fractions containing the Golgi apparatus and the endosomal/lysosomal compartments. Syntaxin 8 in membrane extract could be incorporated into a 20S protein complex in a way that is dependent on the soluble N-ethylmaleimide-sensitive factor (NSF) and soluble NSF attachment protein ((alpha)-SNAP), suggesting that syntaxin 8 is indeed a SNAP receptor (SNARE). Indirect immunofluorescence microscopy reveals that the majority of syntaxin 8 is localized to the early endosome marked by Rab5. This is corroborated by immunogold labeling experiments showing enrichment of syntaxin 8 in the early endosome and its co-labeling with Rab5. PMID- 10683149 TI - Interactions between Fc(epsilon)RI and lipid raft components are regulated by the actin cytoskeleton. AB - Previous studies showed that crosslinking of IgE-Fc(epsilon)RI complexes on RBL 2H3 mast cells causes their association with isolated detergent-resistant membranes, also known as lipid rafts, in a cholesterol-dependent process that precedes initiation of signaling by these receptors. To investigate these interactions on intact cells, we examined the co-redistribution of raft components with crosslinked IgE-Fc(epsilon)RI using confocal microscopy. After several hours of crosslinking at 4 degrees C, the glycosylphosphatidylinositol linked protein Thy-1 and the Src-family tyrosine kinase Lyn co-redistribute with IgE-Fc(epsilon)RI in large patches at the plasma membrane. Under these conditions, F-actin also undergoes dramatic co-segregation with Fc(epsilon)RI and raft components but is dispersed following a brief warm-up to 37 degrees C. When crosslinking of IgE-Fc(epsilon)RI is initiated at higher temperatures, co redistribution of raft components with patched Fc(epsilon)RI is not readily detected unless stimulated F-actin polymerization is inhibited by cytochalasin D. In parallel, cytochalasin D converts transient antigen-stimulated tyrosine phosphorylation to a more sustained response. Sucrose gradient analysis of lysed cells reveals that crosslinked IgE-Fc(epsilon)RI remains associated with lipid rafts throughout the time course of the transient phosphorylation response but undergoes a time-dependent shift to higher density that is prevented by cytochalasin D. Our results indicate that interactions between Lyn and crosslinked IgE-Fc(epsilon)RI are regulated by stimulated F-actin polymerization, and this is best explained by a segregation of anchored raft components from more mobile ones. PMID- 10683150 TI - Endo180, an endocytic recycling glycoprotein related to the macrophage mannose receptor is expressed on fibroblasts, endothelial cells and macrophages and functions as a lectin receptor. AB - Endo180 was previously characterized as a novel, cell type specific, recycling transmembrane glycoprotein. This manuscript describes the isolation of a full length human Endo180 cDNA clone which was shown to encode a fourth member of a family of proteins comprising the macrophage mannose receptor, the phospholipase A(2) receptor and the DEC-205/MR6 receptor. This receptor family is unusual in that they contain 8-10 C-type lectin carbohydrate recognition domains in a single polypeptide backbone, however, only the macrophage mannose receptor had been shown to function as a lectin. Sequence analysis of Endo180 reveals that the second carbohydrate recognition domain has retained key conserved amino acids found in other functional C-type lectins. Furthermore, it is demonstrated that this protein displays Ca(2+)-dependent binding to N-acetylglucosamine but not mannose affinity columns. In order to characterize the physiological function of Endo180, a series of biochemical and morphological studies were undertaken. Endo180 is found to be predominantly expressed in vivo and in vitro on fibroblasts, endothelial cells and macrophages, and the distribution and post translational processing in these cells is consistent with Endo180 functioning to internalize glycosylated ligands from the extracellular milieu for release in an endosomal compartment. PMID- 10683151 TI - Evidence for the coincident initiation of homolog pairing and synapsis during the telomere-clustering (bouquet) stage of meiotic prophase. AB - To improve knowledge of the prerequisites for meiotic chromosome segregation in higher eukaryotes, we analyzed the spatial distribution of a pair of homologs before and during early meiotic prophase. Three-dimensional images of fluorescence in situ hybridization (FISH) were used to localize a single pair of homologs in diploid nuclei of a chromosome-addition line of oat, oat-maize9b. The system provided a robust assay for pairing based on cytological colocalization of FISH signals. Using a triple labeling scheme for simultaneous imaging of chromatin, telomeres and the homolog pair, we determined the timing of pairing in relation to the onset of three sequential hallmarks of early meiotic prophase: chromatin condensation (the leptotene stage), meiotic telomere clustering (the bouquet stage) and the initiation of synapsis (the zygotene stage). We found that the two homologs were mostly unpaired up through middle leptotene, at which point their spherical cloud-like domains began to transform into elongated and stretched-out domains. At late leptotene, the homologs had completely reorganized into long extended fibers, and the beginning of the bouquet stage was conspicuously marked by the de novo clustering of telomeres at the nuclear periphery. The homologs paired and synapsed during the bouquet stage, consistent with the timing of pairing observed for several oat 5S rDNA loci. In summary, results from analysis of more than 100 intact nuclei lead us to conclude that pairing and synapsis of homologous chromosomes are largely coincident processes, ruling out a role for premeiotic pairing in this system. These findings suggest that the genome-wide remodeling of chromatin and telomere-mediated nuclear reorganization are prerequisite steps to the DNA sequence-based homology-search process in higher eukaryotes. PMID- 10683152 TI - The transmembrane protein p23 contributes to the organization of the Golgi apparatus. AB - In previous studies we have shown that p23, a member of the p24-family of small transmembrane proteins, is highly abundant in membranes of the cis-Golgi network (CGN), and is involved in sorting/trafficking in the early secretory pathway. In the present study, we have further investigated the role of p23 after ectopic expression. We found that ectopically expressed p23 folded and oligomerized properly, even after overexpression. However, in contrast to endogenous p23, exogenous p23 molecules did not localize to the CGN, but induced a significant expansion of characteristic smooth ER membranes, where they accumulated in high amounts. This ER-derived, p23-rich subdomain displayed a highly regular morphology, consisting of tubules and/or cisternae of constant diameter, which were reminiscent of the CGN membranes containing p23 in control cells. The expression of exogenous p23 also led to the specific relocalization of endogenous p23, but not of other proteins, to these specialized ER-derived membranes. Relocalization of p23 modified the ultrastructure of the CGN and Golgi membranes, but did not affect anterograde and retrograde transport reactions to any significant extent. We conclude (i) that p23 has a morphogenic activity that contributes to the morphology of CGN-membranes; and (ii) that the presence of p23 in the CGN is necessary for the proper organization of the Golgi apparatus. PMID- 10683153 TI - Brain-derived neurotrophic factor-induced phosphorylation of neurofilament-H subunit in primary cultures of embryo rat cortical neurons. AB - Phosphorylation of the neurofilament-H subunit (NF-H) was investigated in rat embryonic brain neurons in culture. A portion of the NF-H was phosphorylated in vivo at embryonic day 17 when brain neurons were prepared. When the neurons were isolated and cultured, the NF proteins disappeared once and then reappeared over the next several days in the following order: (1) NF-L/NF-M, (2) dephosphorylated NF-H and (3) phosphorylated NF-H. Phosphorylation of NF-H began around 4 days after cell plating, at about the time of synapse formation. Treatments that appeared to modulate the timing of synapse formation also affected the timing of NF-H phosphorylation: (1) earlier phosphorylation was observed at higher neuronal cell density, (2) earlier phosphorylation was observed in neurons cultured on a coating substrate that promotes rapid neurite extension and (3) phosphorylation was suppressed when neurite extension was inhibited by brefeldin A. Three possible synapse formation-induced events, excitation, cell-cell contact through adhesion proteins and elevated concentrations of neurotrophic factors, were examined for their possible involvement in generating the signal for NF-H phosphorylation. Neither excitation nor cell contact enhanced NF-H phosphorylation. Neurotrophic factors, brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) stimulated phosphorylation of NF-H. The BDNF-stimulated phosphorylation was inhibited by an anti-BDNF antibody and K252a, an inhibitor of BDNF receptor TrkB tyrosine kinase. Among known NF-H kinases of cyclin-dependent kinase 5 (CDK5), external signal-regulated protein kinase (ERK) and stress activated protein kinase (SAPK), CDK5 and SAPK showed an increase in kinase activity or an active form with a time course similar to NF-H phosphorylation in control culture. On the other hand, BDNF stimulated the kinase activity of CDK5 and induced appearance of an active form of ERK transiently. These results suggest a possibility that synapse formation induces NF-H phosphorylation, at least in part, through activation of CDK5 by BDNF. PMID- 10683154 TI - Evidence for deterministic chaos in aperiodic oscillations of proliferative activity in long-term cultured Fao hepatoma cells. AB - The proliferative activity of long-term cultured mammalian cells exhibits traits of a complex dynamic system, with a succession of spontaneous rises and falls in proliferation rate. We analyzed three successive series of proliferation data for the Fao hepatoma cell line in long-term cultures. In the three series the proliferation rate displayed apparently disordered oscillations, which each lasted about 3-5 passages, with variable amplitude and were therefore unpredictable. Such non-linear kinetics raises the major issue of whether these fluctuations are random, or determined and coordinated. We used a graphical method of analysis of the data, which demonstrated that all troughs of proliferation were mathematically related to a common value in each series. This common value was itself related to the maximum level of proliferation of the cell line. Non-linear analysis thus confirmed that the fluctuations in proliferation rate of tumoral Fao cells are, at least in part, determined. This pattern evokes chaotic dynamics and is evidence for the flexible coordination of the complex system linking positive and negative growth regulators in long-term cultured cells. PMID- 10683155 TI - A novel mutant allele of the chromatin-bound fission yeast checkpoint protein Rad17 separates the DNA structure checkpoints. AB - To further dissect the genetic differences between the checkpoint pathway following S-phase cdc arrest versus DNA damage, a genetic screen was performed for checkpoint mutants that were unable to arrest mitosis following cell-cycle arrest with a temperature-sensitive DNA polymerase delta mutant, cdc20-M10. One such checkpoint mutant, rad17-d14, was found to display the cut phenotype following S-phase arrest by cdc20-M10, but not by the DNA synthesis inhibitor hydroxyurea, reminiscent of the chk1 mutant. Unlike chk1 , rad17-d14 was not sensitive to UV irradiation. Interestingly, the ionising radiation sensitivity of rad17-d14 was only at higher doses, and cells were found to be defective in properly arresting cell division following irradiation in S phase, but not G(2) phase. Biochemical analysis attributes the checkpoint defects of rad17-d14 to the failure to phosphorylate the checkpoint effector Chk1p. To investigate if Rad17p monitors the genome for abnormal DNA structures specifically during DNA synthesis, chromatin association of Rad17p was analysed. Rad17p was found to be chromatin associated throughout the cell cycle, not just during S phase. This interaction occurred irrespective of the arrest with cdc20-M10 and, surprisingly, was also independent of the other checkpoint Rad proteins, and the cell-cycle effectors Chk1p and Cds1p. PMID- 10683156 TI - Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides. AB - In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression. Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase. This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation. PMID- 10683157 TI - Analysis of the three-dimensional trajectories of organisms: estimates of velocity, curvature and torsion from positional information. AB - Most biological motions are three-dimensional. This includes the trajectories of whole organisms and of their appendages. While recordings of three-dimensional trajectories are sometimes published, quantitative analysis of these trajectories is uncommon, primarily because there are no standard techniques or conventions in biology for the analysis of three-dimensional trajectories. This paper describes a new technique, finite helix fit (FHF), based on the geometry of three dimensional curves, whereby a three-dimensional trajectory is completely described by its velocity, curvature and torsion. FHF estimates these parameters from discretely sampled points on a trajectory (i.e. from positional data such as x,y,z coordinates). Other measures of motion can be derived from these parameters, such as the translational and rotational (or angular) velocities of an organism. The performance of the algorithms is demonstrated using simulated trajectories and trajectories of freely swimming organisms (a flagellate, Chlamydomonas reinhardtii; a ciliate, Paramecium tetraurelia; spermatozoa of a sea urchin, Arbacia punctulata; larvae of an ascidian, Botrylloides sp.). PMID- 10683158 TI - Directional hearing of a grasshopper in the field. AB - An electrophysiological method for making long-term recordings from the tympanal nerve was developed in Chorthippus biguttulus (Gomphocerinae) to gain insight into the ecophysiological constraints of sound localization in acridid grasshoppers. Using this 'biological microphone', the directional dependence of auditory nerve activity was monitored both in the laboratory and in various natural habitats of this species. On gravel and in sparse vegetation, the overall patterns of directionality were found to be very similar to those in the free sound field in the laboratory, regardless of whether the animal was positioned horizontally or vertically. However, the differences between the ipsi- and contralateral sides were smaller in these habitats than in the laboratory. In dense vegetation, the directional patterns were greatly affected by the environment. Moreover, a minimum in nerve activity was not always reached on the contralateral side, as is typical for the free sound field situation. On the basis of these data, predictions can be made about the ability of the animals to determine the correct side of a sound source. In the free sound field of the laboratory, correct lateralizations are expected at all angles of sound incidence between 20 and 160 degrees, a prediction corresponding to the results of behavioural studies. In sparse vegetation, a similar accuracy can be anticipated, whereas on gravel and in dense vegetation directional hearing is expected to be severely degraded, especially if the animal is oriented horizontally. The predictions from our present electrophysiological investigations must now be confirmed by behavioural studies in the field. PMID- 10683159 TI - Lack of effect of ageing on acetate oxidation in rat skeletal muscle during starvation: a (13)C NMR study. AB - Acetate oxidation was examined by (13)C nuclear magnetic resonance in skeletal muscle from adult and old rats. Rats fasted for 5 days were perfused with [2 (13)C]acetate over 2 h, and muscle extracts were analyzed for [(13)C]glutamate isotopomers. This study shows that approximately 80 % of acetyl-coenzyme A entering the tricarboxylic cycle was derived from substrate infusion in both adult and old rats, and that the flux through anaplerotic pathways was approximately 21 % of the flux through citrate synthase. These data demonstrate that skeletal muscle from adult and old rats oxidizes the same proportion of exogenous acetate. PMID- 10683160 TI - The ontogeny of metabolic rate and thermoregulatory capabilities of northern fur seal, Callorhinus ursinus, pups in air and water. AB - Young pinnipeds, born on land, must eventually enter the water to feed independently. The aim of this study was to examine developmental factors that might influence this transition. The ontogeny of metabolic rate and thermoregulation in northern fur seal, Callorhinus ursinus, pups was investigated at two developmental stages in air and water using open-circuit respirometry. Mean in-air resting metabolic rate (RMR) increased significantly from 113+/-5 ml O(2 )min(-)(1) (N=18) pre-molt to 160+/-4 ml O(2 )min(-)(1) (N=16; means +/- s.e.m.) post-molt. In-water, whole-body metabolic rates did not differ pre- and post-molt and were 2.6 and 1.6 times in-air RMRs respectively. Mass-specific metabolic rates of pre-molt pups in water were 2.8 times in-air rates. Mean mass specific metabolic rates of post-molt pups at 20 degrees C in water and air did not differ (16.1+/-1.7 ml O(2 )min(-)(1 )kg(-)(1); N=10). In-air mass-specific metabolic rates of post-molt pups were significantly lower than in-water rates at 5 degrees C (18.2+/-1.1 ml O(2 )min(-)(1 )kg(-)(1); N=10) and 10 degrees C (19.4+/-1.7 ml O(2 )min(-)(1 )kg(-)(1); N=10; means +/- s.e.m.). Northern fur seal pups have metabolic rates comparable with those of terrestrial mammalian young of similar body size. Thermal conductance was independent of air temperature, but increased with water temperature. In-water thermal conductance of pre-molt pups was approximately twice that of post-molt pups. In-water pre molt pups matched the energy expenditure of larger post-molt pups while still failing to maintain body temperature. Pre-molt pups experience greater relative costs when entering the water regardless of temperature than do larger post-molt pups. This study demonstrates that the development of thermoregulatory capabilities plays a significant role in determining when northern fur seal pups enter the water. PMID- 10683161 TI - Thermotolerant desert lizards characteristically differ in terms of heat-shock system regulation. AB - We compare the properties and activation of heat-shock transcription factor (HSF1) and the synthesis of a major family of heat-shock proteins (HSP70) in lizard species inhabiting ecological niches with strikingly different thermal parameters. Under normal non-heat-shock conditions, all desert-dwelling lizard species studied so far differ from a northern, non-desert species (Lacerta vivipara) in the electrophoretic mobility and content of proteins constitutively bound to the regulatory heat-shock elements in the heat-shock gene promoter. Under these conditions, levels of activated HSF1 and of both HSP70 mRNA and protein are higher in the desert species than in the non-desert species. Upon heat shock, HSF1 aggregates in all species studied, although in desert species HSF1 subsequently disaggregates more rapidly. Cells of the northern species have a lower thermal threshold for HSP expression than those of the desert species, which correlates with the relatively low constitutive level of HSPs and high basal content of HSF1 in their cells. PMID- 10683162 TI - Differential branchial and renal handling of urea, acetamide and thiourea in the gulf toadfish Opsanus beta: evidence for two transporters. AB - The possible presence of a urea transporter in the kidney of the gulf toadfish (Opsanus beta) and further characterization of the pulsatile facilitated transporter previously identified in its gills were investigated by comparing the extra-renal and renal handling of two urea analogues with the handling of urea. Toadfish were fitted with caudal artery and indwelling urinary ureteral catheters and injected with an iso-osmotic dose of (14)C-labelled urea analogue (acetamide or thiourea) calculated to bring plasma analogue concentrations close to plasma urea concentrations. Branchial permeabilities to urea, acetamide and thiourea were similar during non-pulsing periods and all increased during pulse events, although urea permeability was greater than analogue permeability during pulses. The incidence and magnitude of acetamide and urea pulses at the gills were significantly correlated, acetamide pulses being 35-50 % of the size of urea pulses. However, the thiourea and urea pulses at the gills were only weakly correlated, thiourea pulses being less than 16 % of the size of urea pulses. Thiourea inhibited branchial urea excretion by reducing the pulse frequency. The renal handling of thiourea and urea were similar in that both substances were more concentrated in the urine than in the plasma, whereas acetamide was found in equal concentrations in the urine and plasma. Urea and thiourea were secreted 2-3 times more effectively than Cl(-) and water, whereas acetamide was secreted at a similar relative rate. The differential handling of the urea analogues by the gills and kidney indicates the presence of a different, possibly unique, transporter in the kidney. The movement of thiourea and urea into the renal tubule against an apparent concentration gradient suggests the presence of an active transport mechanism. PMID- 10683163 TI - The effects of cell ageing on metabolism in rainbow trout (Oncorhynchus mykiss) red blood cells. AB - The effects of cell age on metabolism in the nucleated red blood cells of rainbow trout (Oncorhynchus mykiss) were examined. Red blood cells were separated according to age using fixed-angle centrifugation. The mean erythrocyte haemoglobin concentration in old red blood cells was found to be 120 % of that in young red blood cells. In young red blood cells, the activities of the mitochondrial enzymes citrate synthase and cytochrome oxidase were 135-200 %, respectively, of those measured in old red blood cells. The activity of the glycolytic enzyme lactate dehydrogenase in young red blood cells was 170 % of that in old red blood cells, whereas the activity of the glycolytic enzyme pyruvate kinase was not significantly affected by cell age. In addition, young red blood cells consumed over twice as much O(2) and devoted 50 % more O(2) to protein synthesis and the activity of Na(+)/K(+)-ATPase than old red blood cells. Red blood cell age did not significantly affect the rate of lactate production. This study shows that ageing in rainbow trout nucleated red blood cells is accompanied by a significant decline in aerobic energy production and the processes it supports, as well as a corresponding increase in the glycolytic contribution to metabolism. PMID- 10683164 TI - Sulphaemoglobin formation in fish: a comparison between the haemoglobin of the sulphide-sensitive rainbow trout (Oncorhynchus Mykiss) and of the sulphide tolerant common carp (Cyprinus Carpio). AB - A method for the quantitative determination of sulphaemoglobin (SHb) in a mixture of haemoglobin derivatives by spectral deconvolution is described. SHb formation was studied in haemolysates and in red blood cells of the sulphide-sensitive rainbow trout (Oncorhynchus mykiss) and of the sulphide-tolerant common carp (Cyprinus carpio). Addition of sulphide caused the formation of SHb in haemolysates of both animals. However, haemoglobin from common carp was much less sensitive to sulphide than was trout haemoglobin. The maximal obtainable SHb fraction was approximately 30 % in trout and 10 % in carp haemolysates. In both animals, the SHb fraction increased with increasing Hb and sulphide concentrations up to 100 micromol l(-)(1) and 1 mmol l(-)(1), respectively, and was favoured by a low pH. An increase of temperature between 5 and 25 degrees C strongly increased SHb formation in trout haemolysate. In contrast, temperature changes had almost no effect on SHb production in carp. Within trout red blood cells, approximately 7 % of total haemoglobin was converted to SHb during 60 min of incubation (with 2.5 mmol l(-)(1) sulphide), inducing a 20 % loss of haemoglobin oxygen-saturation. In carp red blood cells incubated under identical conditions, SHb formation was minimal and haemoglobin oxygen-saturation was not affected. PMID- 10683165 TI - Physical and physiological components of the graviresponses of wild-type and mutant Paramecium Tetraurelia. AB - Wild-type and the morphological mutant kin 241 of Paramecium tetraurelia showed improved orientation away from the centre of gravity (negative gravitaxis) when accelerations were increased from 1 to 7 g. Gravitaxis was more pronounced in the mutant. A correlation between the efficiency of orientation and the applied g value suggests a physical basis for gravitaxis. Transiently enhanced rates of reversal of the swimming direction coincided with transiently enhanced gravitaxis because reversals occurred more often in downward swimmers than in upward swimmers. The results provide evidence of a physiological modulation of gravitaxis by means of the randomizing effect of depolarization-dependent swimming reversals. Gravity bimodally altered propulsion rates of wild-type P. tetraurelia so that sedimentation was partly antagonized in upward and downward swimmers (negative gravikinesis). In the mutant, only increases in propulsion were observed, although the orientation-dependent sensitivity of the gravikinetic response was the same as in the wild-type population. Observed swimming speed and sedimentation rates in the wild-type and mutant cells were linearly related to acceleration, allowing the determination of gravikinesis as a linear (and so far non-saturating) function of gravity. PMID- 10683166 TI - A conserved location for the central nervous system control of mating behaviour in gastropod molluscs: evidence from a terrestrial snail. AB - We have investigated the role of the right mesocerebrum in the expression of mating behaviour in the garden snail Helix aspersa. Using an in vivo stimulation and recording technique, we provide evidence for both sensory and motor functions in the mesocerebral neuronal population. Some neurones were specifically sensitive to tactile stimuli delivered to the skin on the superior tentacles and around the genital pore. Electrical stimulation of the right mesocerebrum evoked genital eversion and, in combination with tactile stimulation, dart-shooting and penial eversion. Genital eversions were also elicited by injections of APGWamide. During courtship, one recorded unit increased its activity only in correlation with penial eversion, while six other units increased their activity only during dart-shooting. Three additional units increased their activity during both types of behaviour. In addition, most of the recorded units showed increased neuronal activity during times of contact with a partner. Comparison of our results with available data from other molluscs leads us to conclude that the right anteromedial region of the cerebral ganglion is an evolutionarily conserved region of the gastropod brain specialised for the control of male mating behaviour. It is striking to find such functional conservation in the central nervous system of phylogenetically distant gastropods given the large differences in behaviour during mating. PMID- 10683167 TI - Mechanics of lung ventilation in a post-metamorphic salamander, Ambystoma Tigrinum. AB - The mechanics of lung ventilation in frogs and aquatic salamanders has been well characterized, whereas lung ventilation in terrestrial-phase (post-metamorphic) salamanders has received little attention. We used electromyography (EMG), X-ray videography, standard videography and buccal and body cavity pressure measurements to characterize the ventilation mechanics of adult (post metamorphic) tiger salamanders (Ambystoma tigrinum). Three results emerged: (i) under terrestrial conditions or when floating at the surface of the water, adult A. tigrinum breathed through their nares using a two-stroke buccal pump; (ii) in addition to this narial two-stroke pump, adult tiger salamanders also gulped air in through their mouths using a modified two-stroke buccal pump when in an aquatic environment; and (iii) exhalation in adult tiger salamanders is active during aquatic gulping breaths, whereas exhalation appears to be passive during terrestrial breathing at rest. Active exhalation in aquatic breaths is indicated by an increase in body cavity pressure during exhalation and associated EMG activity in the lateral hypaxial musculature, particularly the M. transversus abdominis. In terrestrial breathing, no EMG activity in the lateral hypaxial muscles is generally present, and body cavity pressure decreases during exhalation. In aquatic breaths, tidal volume is larger than in terrestrial breaths, and breathing frequency is much lower (approximately 1 breath 10 min( )(1 )versus 4-6 breaths min(-)(1)). The use of hypaxial muscles to power active exhalation in the aquatic environment may result from the need for more complete exhalation and larger tidal volumes when breathing infrequently. This hypothesis is supported by previous findings that terrestrial frogs ventilate their lungs with small tidal volumes and exhale passively, whereas aquatic frogs and salamanders use large tidal volumes and and exhale actively. PMID- 10683168 TI - The electrical properties of the anterior stomach of the larval mosquito (Aedes aegypti). AB - The electrical properties of the anterior stomach of the larval mosquito (Aedes aegypti) were determined. At late times after cannulation, the intraluminal space constant was 936 microm, which is almost as long as the isolated tissue itself. At this time, the resistance of the apical cell membranes dominates the transcellular resistance; it is approximately 14 times the resistance of the basal cell membrane. Two physiologically distinct epithelial cell types were identified. One type has a stable basal potential of approximately 65 mV and responds to 5-hydroxytryptamine with hyperpolarization. The second cell type initially shows a basal potential of 100 mV. However, this basal potential decays in the first few minutes in parallel with the decay of the transintestinal potential. This latter cell type does not respond to 5-hydroxytryptamine. PMID- 10683169 TI - The RXR ortholog USP suppresses early metamorphic processes in Drosophila in the absence of ecdysteroids. AB - The steroid hormone 20-hydroxyecdysone (20E) initiates metamorphosis in insects by signaling through the ecdysone receptor complex, a heterodimer of the ecdysone receptor (EcR) and ultraspiracle (USP). Analysis of usp mutant clones in the wing disc of Drosophila shows that in the absence of USP, early hormone responsive genes such as EcR, DHR3 and E75B fail to up-regulate in response to 20E, but other genes that are normally expressed later, such as (&bgr;)-Ftz-F1 and the Z1 isoform of the Broad-Complex (BRC-Z1), are expressed precociously. Sensory neuron formation and axonal outgrowth, two early metamorphic events, also occur prematurely. In vitro experiments with cultured wing discs showed that BRC-Z1 expression and early metamorphic development are rendered steroid-independent in the usp mutant clones. These results are consistent with a model in which these latter processes are induced by a signal arising during the middle of the last larval stage but suppressed by the unliganded EcR/USP complex. Our observations suggest that silencing by the unliganded EcR/USP receptor and the subsequent release of silencing by moderate steroid levels may play an important role in coordinating early phases of steroid driven development. PMID- 10683170 TI - In ovo time-lapse analysis of chick hindbrain neural crest cell migration shows cell interactions during migration to the branchial arches. AB - Hindbrain neural crest cells were labeled with DiI and followed in ovo using a new approach for long-term time-lapse confocal microscopy. In ovo imaging allowed us to visualize neural crest cell migration 2-3 times longer than in whole embryo explant cultures, providing a more complete picture of the dynamics of cell migration from emergence at the dorsal midline to entry into the branchial arches. There were aspects of the in ovo neural crest cell migration patterning which were new and different. Surprisingly, there was contact between neural crest cell migration streams bound for different branchial arches. This cell-cell contact occurred in the region lateral to the otic vesicle, where neural crest cells within the distinct streams diverted from their migration pathways into the branchial arches and instead migrated around the otic vesicle to establish a contact between streams. Some individual neural crest cells did appear to cross between the streams, but there was no widespread mixing. Analysis of individual cell trajectories showed that neural crest cells emerge from all rhombomeres (r) and sort into distinct exiting streams adjacent to the even-numbered rhombomeres. Neural crest cell migration behaviors resembled the wide diversity seen in whole embryo chick explants, including chain-like cell arrangements; however, average in ovo cell speeds are as much as 70% faster. To test to what extent neural crest cells from adjoining rhombomeres mix along migration routes and within the branchial arches, separate groups of premigratory neural crest cells were labeled with DiI or DiD. Results showed that r6 and r7 neural crest cells migrated to the same spatial location within the fourth branchial arch. The diversity of migration behaviors suggests that no single mechanism guides in ovo hindbrain neural crest cell migration into the branchial arches. The cell-cell contact between migration streams and the co-localization of neural crest cells from adjoining rhombomeres within a single branchial arch support the notion that the pattern of hindbrain neural crest cell migration emerges dynamically with cell cell communication playing an important guidance role. PMID- 10683172 TI - Autoregulation and multiple enhancers control Math1 expression in the developing nervous system. AB - Development of the vertebrate nervous system requires the actions of transcription factors that establish regional domains of gene expression, which results in the generation of diverse neuronal cell types. MATH1, a transcription factor of the bHLH class, is expressed during development of the nervous system in multiple neuronal domains, including the dorsal neural tube, the EGL of the cerebellum and the hair cells of the vestibular and auditory systems. MATH1 is essential for proper development of the granular layer of the cerebellum and the hair cells of the cochlear and vestibular systems, as shown in mice carrying a targeted disruption of Math1. Previously, we showed that 21 kb of sequence flanking the Math1-coding region is sufficient for Math1 expression in transgenic mice. Here we identify two discrete sequences within the 21 kb region that are conserved between mouse and human, and are sufficient for driving a lacZ reporter gene in these domains of Math1 expression in transgenic mice. The two identified enhancers, while dissimilar in sequence, appear to have redundant activities in the different Math1 expression domains except the spinal neural tube. The regulatory mechanisms for each of the diverse Math1 expression domains are tightly linked, as separable regulatory elements for any given domain of Math1 expression were not found, suggesting that a common regulatory mechanism controls these apparently unrelated domains of expression. In addition, we demonstrate a role for autoregulation in controlling the activity of the Math1 enhancer, through an essential E-box consensus binding site. PMID- 10683171 TI - Endodermal Nodal-related signals and mesoderm induction in Xenopus. AB - In Xenopus, mesoderm induction by endoderm at the blastula stage is well documented, but the molecular nature of the endogenous inductive signals remains unknown. The carboxy-terminal fragment of Cerberus, designated Cer-S, provides a specific secreted antagonist of mesoderm-inducing Xenopus Nodal-Related (Xnr) factors. Cer-S does not inhibit signalling by other mesoderm inducers such as Activin, Derriere, Vg1 and BMP4, nor by the neural inducer Xnr3. In the present study we show that Cer-S blocks the induction of both dorsal and ventral mesoderm in animal-vegetal Nieuwkoop-type recombinants. During blastula stages Xnr1, Xnr2 and Xnr4 are expressed in a dorsal to ventral gradient in endodermal cells. Dose response experiments using cer-S mRNA injections support the existence of an endogenous activity gradient of Xnrs. Xnr expression at blastula can be activated by the vegetal determinants VegT and Vg1 acting in synergy with dorsal (beta) catenin. The data support a modified model for mesoderm induction in Xenopus, in which mesoderm induction is mediated by a gradient of multiple Nodal-related signals released by endoderm at the blastula stage. PMID- 10683173 TI - Fragile skeletal muscle attachments in dystrophic mutants of Caenorhabditis elegans: isolation and characterization of the mua genes. AB - Over 30 Caenorhabditis elegans mutants were identified with normal muscle differentiation and initial locomotion followed by catastrophic detachment of skeletal muscles from the body wall. Reducing the strength of muscle contraction in these mutants with a myosin gene mutation suppresses muscle detachment. These dystrophic mutants identify a novel class of genes required for growth and maintenance of functional muscle attachments, not exceptional alleles of genes required for muscle differentiation and contractility. Nine new genes, named mua, and two previously published loci, unc-23 and vab-10, cause fragile musscle attachments. The primary sites of muscle detachment, including the plane of tissue separation, are characteristic for each gene. We suggest these genes identify feedback mechanisms whereby local strain regulates the extent of myofibril contraction and the placement of new muscle attachments in functioning muscles. Finally, we draw some comparisons to vertebrate skin fragility diseases and muscular dystrophies. PMID- 10683174 TI - Dorsal and intermediate neuronal cell types of the spinal cord are established by a BMP signaling pathway. AB - We have studied the role of Bmp signaling in patterning neural tissue through the use of mutants in the zebrafish that disrupt three different components of a Bmp signaling pathway: swirl/bmp2b, snailhouse/bmp7 and somitabun/smad5. We demonstrate that Bmp signaling is essential for the establishment of the prospective neural crest and dorsal sensory Rohon-Beard neurons of the spinal cord. Moreover, Bmp signaling is necessary to limit the number of intermediate positioned lim1+ interneurons of the spinal cord, as observed by the dramatic expansion of these prospective interneurons in many mutant embryos. Our analysis also suggests a positive role for Bmp signaling in the specification of these interneurons, which is independent of Bmp2b/Swirl activity. We found that a presumptive ventral signal, Hh signaling, acts to restrict the amount of dorsal sensory neurons and trunk neural crest. This restriction appears to occur very early in neural tissue development, likely prior to notochord or floor plate formation. A similar early role for Bmp signaling is suggested in the specification of dorsal neural cell types, since the bmp2b/swirl and bmp7/snailhouse genes are only coexpressed during gastrulation and within the tail bud, and are not found in the dorsal neural tube or overlying epidermal ectoderm. Thus, a gastrula Bmp2b/Swirl and Bmp7/Snailhouse-dependent activity gradient may not only act in the specification of the embryonic dorsoventral axis, but may also function in establishing dorsal and intermediate neuronal cell types of the spinal cord. PMID- 10683175 TI - Regulation of the early expression of the Xenopus nodal-related 1 gene, Xnr1. AB - The Xenopus nodal related-1 (Xnr1) gene has a complex expression pattern in embryos, with two temporal phases. In the first phase, transcripts are first detected in perinuclear sites in the vegetal region of the blastula. During gastrulation, this expression disappears and transcripts become localised to the dorsal marginal zone. Expression stops and then restarts in a second phase at neurula and tailbud stages, firstly in two symmetric patches near the posterior end of the notochord, and then asymmetrically in a large domain in the left lateral plate mesoderm. In this study, we have investigated the regulation of the early phase of expression of Xnr1. We show that the T-box transcription factor VegT can induce Xnr1. It had previously been shown that Xnr1 can induce VegT in ectoderm cells and we show that the early expression of Xnr1 is regulated by an autoregulatory loop. By inspection of the Xnr1 promoter sequence, we have identified two non-palindromic T-box-binding sites, which are 10 bp apart. Using mutational analysis, we have shown that these elements are required for the VegT induction of Xnr1. The Xnr1 promoter shows striking homologies with the Xnr3 promoter. In particular, two elements that are required for Wnt signaling are conserved between these two promoters, but the two T-box sites are not conserved, and Xnr3 is not induced by VegT. A region of the promoter containing the T-box sites and the Wnt sites is sufficient to drive expression of a reporter gene in a dorsal domain in transgenic Xenopus at the gastrula stage. We show that this pattern of expression of the transgene in gastrulae is not dependent on the T-box sites. PMID- 10683176 TI - Kinase independent function of EphB receptors in retinal axon pathfinding to the optic disc from dorsal but not ventral retina. AB - Optic nerve formation requires precise retinal ganglion cell (RGC) axon pathfinding within the retina to the optic disc, the molecular basis of which is not well understood. At CNS targets, interactions between Eph receptor tyrosine kinases on RGC axons and ephrin ligands on target cells have been implicated in formation of topographic maps. However, studies in chick and mouse have shown that both Eph receptors and ephrins are also expressed within the retina itself, raising the possibility that this receptor-ligand family mediates aspects of retinal development. Here, we more fully document the presence of specific EphB receptors and B-ephrins in embryonic mouse retina and provide evidence that EphB receptors are involved in RGC axon pathfinding to the optic disc. We find that as RGC axons begin this pathfinding process, EphB receptors are uniformly expressed along the dorsal-ventral retinal axis. This is in contrast to the previously reported high ventral-low dorsal gradient of EphB receptors later in development when RGC axons map to CNS targets. We show that mice lacking both EphB2 and EphB3 receptor tyrosine kinases, but not each alone, exhibit increased frequency of RGC axon guidance errors to the optic disc. In these animals, major aspects of retinal development and cellular organization appear normal, as do the expression of other RGC guidance cues netrin, DCC, and L1. Unexpectedly, errors occur in dorsal but not ventral retina despite early uniform or later high ventral expression of EphB2 and EphB3. Furthermore, embryos lacking EphB3 and the kinase domain of EphB2 do not show increased errors, consistent with a guidance role for the EphB2 extracellular domain. Thus, while Eph kinase function is involved in RGC axon mapping in the brain, RGC axon pathfinding within the retina is partially mediated by EphB receptors acting in a kinase-independent manner. PMID- 10683177 TI - Completion of meiosis in Drosophila oocytes requires transcriptional control by grauzone, a new zinc finger protein. AB - Mutations in grauzone or cortex cause abnormal arrest in Drosophila female meiosis. We cloned grauzone and identified it as a C2H2-type zinc finger transcription factor. The grauzone transcript is present in ovaries and at later developmental stages. A Grauzone-GFP fusion protein is functional and localizes to nuclei of both nurse cells and follicle cells during oogenesis. Three lines of evidence indicate that grauzone and cortex interact: reducing cortex function enhanced the grauzone mutant phenotype; cortex transcript abundance is reduced in the absence of grauzone function and Grauzone protein binds to the cortex promoter. These results demonstrate that activation of cortex transcription by grauzone is necessary for the completion of meiosis in Drosophila oocytes, and establish a new pathway that specifically regulates the female meiotic cell cycle. PMID- 10683178 TI - A retrograde signal is involved in activity-dependent remodeling at a C. elegans neuromuscular junction. AB - We have characterized how perturbations of normal synaptic activity influence the morphology of cholinergic SAB motor neurons that innervate head muscle in C. elegans. Mutations disrupting components of the presynaptic release apparatus, acetylcholine (ACh) synthesis or ACh loading into synaptic vesicles each induced sprouting of SAB axonal processes. These sprouts usually arose in the middle of the normal innervation zone and terminated with a single presynaptic varicosity. Sprouting SAB neurons with a similar morphology were also observed upon reducing activity in muscle, either by using mutants lacking a functional nicotinic ACh receptor subunit or through muscle-specific expression of a gain-of-function potassium channel. Analysis of temperature-sensitive mutants in the choline acetyltransferase gene revealed that the sprouting response to inactivity was developmentally regulated; reduction of synaptic activity in early larval stages, but not in late larval stages, induced both sprouting and addition of varicosities. Our results indicate that activity levels regulate the structure of certain synaptic connections between nerve and muscle in C. elegans. One component of this regulatory machinery is a retrograde signal from the postsynaptic cell that mediates the formation of synaptic connections. PMID- 10683179 TI - Regulation of cell proliferation patterns by homeotic genes during Arabidopsis floral development. AB - The shoot apical meristem of Arabidopsis thaliana consists of three cell layers that proliferate to give rise to the aerial organs of the plant. By labeling cells in each layer using an Ac-based transposable element system, we mapped their contributions to the floral organs, as well as determined the degree of plasticity in this developmental process. We found that each cell layer proliferates to give rise to predictable derivatives: the L1 contributes to the epidermis, the stigma, part of the transmitting tract and the integument of the ovules, while the L2 and L3 contribute, to different degrees, to the mesophyll and other internal tissues. In order to test the roles of the floral homeotic genes in regulating these patterns of cell proliferation, we carried out similar clonal analyses in apetala3-3 and agamous-1 mutant plants. Our results suggest that cell division patterns are regulated differently at different stages of floral development. In early floral stages, the pattern of cell divisions is dependent on position in the floral meristem, and not on future organ identity. Later, during organogenesis, the layer contributions to the organs are controlled by the homeotic genes. We also show that AGAMOUS is required to maintain the layered structure of the meristem prior to organ initiation, as well as having a non-autonomous role in the regulation of the layer contributions to the petals. PMID- 10683180 TI - Different clonal dispersion in the rostral and caudal mouse central nervous system. AB - We have performed a systematic clonal analysis to describe the modes of growth, dispersion and production of cells during the development of the mouse neural system. We have used mice expressing a LaacZ reporter gene under the control of the neuron specific enolase promoter to randomly generate LacZ clones in the central nervous system (CNS). We present evidence for (1) a pool of CNS founder cells that is not regionalized, i.e. give descendants dispersed along the entire A-P axis, (2) an early separation between pools of precursors for the anterior and posterior CNS and (3) distinct modes of production of progenitors in these two domains. More specifically, cell growth and dispersion of the progenitors follow a relatively coherent pattern throughout the anterior CNS, a mode that leads to a progressive regionalization of cell fates. In contrast, cell growth of progenitors of the SC appears to involve self-renewing stem cells that progress caudally during regression of the mode. Therefore, at least part of the area surrounding the node is composed of precursors with self-renewing properties and the development of the trunk is dependent on pools of stem cells regressing from A to P. Taken together with our analysis of the cell growth changes associated with neuromere formation (Mathis, L., Sieur, J., Voiculescu, O., Charnay, P. and Nicolas, J. F. (1999) Development 126, 4095-4106), our results suggest that major transitions in CNS development correspond to changes in cell behavior and may provide a link between morphogenesis and genetic patterning mechanisms (i.e. formation of the body plan). PMID- 10683181 TI - The Abruptex domain of Notch regulates negative interactions between Notch, its ligands and Fringe. AB - The Notch signalling pathway regulates cell fate choices during both vertebrate and invertebrate development. In the Drosophila wing disc, the activation of Notch by its ligands Delta and Serrate is required to make the dorsoventral boundary, where several genes, such as wingless and cut, are expressed in a 2- to 4-cell-wide domain. The interactions between Notch and its ligands are modulated by Fringe via a mechanism that may involve post-transcriptional modifications of Notch. The ligands themselves also help to restrict Notch activity to the dorsoventral boundary cells, because they antagonise the activation of the receptor in the cells where their expression is high. This function of the ligands is critical to establish the polarity of signalling, but very little is known about the mechanisms involved in the interactions between Notch and its ligands that result in suppression of Notch activity. The extracellular domain of Notch contains an array of 36 EGF repeats, two of which, repeats 11 and 12, are necessary for direct interactions between Notch with Delta and Serrate. We investigate here the function of a region of the Notch extracellular domain where several missense mutations, called Abruptex, are localised. These Notch alleles are characterised by phenotypes opposite to the loss of Notch function and also by complex complementation patterns. We find that, in Abruptex mutant discs, only the negative effects of the ligands and Fringe are affected, resulting in the failure to restrict the expression of cut and wingless to the dorsoventral boundary. We suggest that Abruptex alleles identify a domain in the Notch protein that mediates the interactions between Notch, its ligands and Fringe that result in suppression of Notch activity. PMID- 10683182 TI - Distinct effects of XBF-1 in regulating the cell cycle inhibitor p27(XIC1) and imparting a neural fate. AB - XBF-1 is an anterior neural plate-specific, winged helix transcription factor that affects neural development in a concentration-dependent manner. A high concentration of XBF-1 results in suppression of endogenous neuronal differentiation and an expansion of undifferentiated neuroectoderm. Here we investigate the mechanism by which this expansion is achieved. Our findings suggest that XBF-1 converts ectoderm to a neural fate and it does so independently of any effects on the mesoderm. In addition, we show that a high dose of XBF-1 promotes the proliferation of neuroectodermal cells while a low dose inhibits ectodermal proliferation. Thus, the neural expansion observed after high dose XBF-1 misexpression is due both to an increase in the number of ectodermal cells devoted to a neural fate and an increase in their proliferation. We show that the effect on cell proliferation is likely to be mediated by p27(XIC1), a cyclin-dependent kinase (cdk) inhibitor. We show that p27(XIC1) is expressed in a spatially restricted pattern in the embryo, including the anterior neural plate, and when misexpressed it is sufficient to block the cell cycle in vivo. We find that p27(XIC1 )is transcriptionally regulated by XBF-1 in a dose dependent manner such that it is suppressed or ectopically induced by a high or low dose of XBF-1, respectively. However, while a low dose of XBF-1 induces ectopic p27(XIC1 )and ectopic neurons, misexpression of p27(XIC1 )does not induce ectopic neurons, suggesting that the effects of XBF-1 on cell fate and cell proliferation are distinct. Finally, we show that p27(XIC1 )is suppressed by XBF 1 in the absence of protein synthesis, suggesting that at least one component of p27(XIC1 )regulation by XBF-1 may be direct. Thus, XBF-1 is a neural-specific transcription factor that can independently affect both the cell fate choice and the proliferative status of the cells in which it is expressed. PMID- 10683183 TI - The neurofibromatosis-2 homologue, Merlin, and the tumor suppressor expanded function together in Drosophila to regulate cell proliferation and differentiation. AB - Neurofibromatosis-2 is an inherited disorder characterized by the development of benign schwannomas and other Schwann-cell-derived tumors associated with the central nervous system. The Neurofibromatosis-2 tumor suppressor gene encodes Merlin, a member of the Protein 4.1 superfamily most closely related to Ezrin, Radixin and Moesin. This discovery suggested a novel function for Protein 4.1 family members in the regulation of cell proliferation; proteins in this family were previously thought to function primarily to link transmembrane proteins to underlying cortical actin. To understand the basic cellular functions of Merlin, we are investigating a Drosophila Neurofibromatosis-2 homologue, Merlin. Loss of Merlin function in Drosophila results in hyperplasia of the affected tissue without significant disruptions in differentiation. Similar phenotypes have been observed for mutations in another Protein 4.1 superfamily member in Drosophila, expanded. Because of the phenotypic and structural similarities between Merlin and expanded, we asked whether Merlin and Expanded function together to regulate cell proliferation. In this study, we demonstrate that recessive loss of function of either Merlin or expanded can dominantly enhance the phenotypes associated with mutations in the other. Consistent with this genetic interaction, we determined that Merlin and Expanded colocalize in Drosophila tissues and cells, and physically interact through a conserved N-terminal region of Expanded, characteristic of the Protein 4.1 family, and the C-terminal domain of Merlin. Loss of function of both Merlin and expanded in clones revealed that these proteins function to regulate differentiation in addition to proliferation in Drosophila. Further genetic analyses suggest a role for Merlin and Expanded specifically in Decapentaplegic-mediated differentiation events. These results indicate that Merlin and Expanded function together to regulate proliferation and differentiation, and have implications for understanding the functions of other Protein 4.1 superfamily members. PMID- 10683184 TI - Morphogenetic furrow initiation and progression during eye development in Drosophila: the roles of decapentaplegic, hedgehog and eyes absent. AB - The Drosophila signaling factor decapentaplegic (dpp) mediates the effects of hedgehog (hh) in tissue patterning by regulating the expression of tissue specific genes. In the eye disc, the transcription factors eyeless (ey), eyes absent (eya), sine oculis (so) and dachshund (dac) participate with these signaling molecules in a complex regulatory network that results in the initiation of eye development. Our analysis of functional relationships in the early eye disc indicates that hh and dpp play no role in regulating ey, but are required for eya, so and dac expression. We show that restoring expression of eya in loss-of-function dpp mutant backgrounds is sufficient to induce so and dac expression and to rescue eye development. Thus, once expressed, eya can carry out its functions in the absence of dpp. These experiments indicate that dpp functions downstream of or in parallel with ey, but upstream of eya, so and dac. Additional control is provided by a feedback loop that maintains expression of eya and so and includes dpp. The fact that exogenous overexpression of ey, eya, so and dac interferes with wild-type eye development demonstrates the importance of such a complicated mechanism for maintaining proper levels of these factors during early eye development. Whereas initiation of eye development fails in either Hh or Dpp signaling mutants, the subsequent progression of the morphogenetic furrow is only slowed down. However, we find that clones that are simultaneously mutant for Hh and Dpp signaling components completely block furrow progression and eye differentiation, suggesting that Hh and Dpp serve partially redundant functions in this process. Interestingly, furrow-associated expression of eya, so and dac is not affected by double mutant tissue, suggesting that some other factor(s) regulates their expression during furrow progression. PMID- 10683185 TI - The pharmacology of hSK1 Ca2+-activated K+ channels expressed in mammalian cell lines. AB - The pharmacology of hSK1, a small conductance calcium-activated potassium channel, was studied in mammalian cell lines (HEK293 and COS-7). In these cell types, hSK1 forms an apamin-sensitive channel with an IC(50) for apamin of 8 nM in HEK293 cells and 12 nM in COS-7 cells. The currents in HEK293 cells were also sensitive to tubocurarine (IC(50)=23 microM), dequalinium (IC(50)=0.4 microM), and the novel dequalinium analogue, UCL1848 (IC(50)=1 nM). These results are very different from the pharmacology of hSK1 channels expressed in Xenopus oocytes and suggest the properties of the channel may depend on the expression system. Our findings also raise questions about the role of SK1 channels in generating the apamin-insensitive slow afterhyperpolarization observed in central neurones. PMID- 10683186 TI - Antihyperglycemic action of isoferulic acid in streptozotocin-induced diabetic rats. AB - Wistar rats with streptozotocin-induced diabetes (STZ-diabetic rats), which is similar to human insulin-dependent diabetic mellitus (IDDM), were employed to investigate the antihyperglycemic action of isoferulic acid. A single intravenous injection of isoferulic acid decreased the plasma glucose in a dose-dependent manner in the STZ-diabetic rats. Repeated intravenous administration of STZ diabetic rats with isoferulic acid (5.0 mg kg(-1)) also resulted in the lowering of plasma glucose after one day. Stimulatory effects of isoferulic acid on the glucose uptake and glycogen synthesis in soleus muscles isolated from STZ diabetic rats were also obtained indicating an increase of glucose utilization following isoferulic acid treatment which was not dependent on insulin. The mRNA level of glucose transporter subtype 4 form (GLUT4) in soleus muscle was raised by isoferulic acid after repeated treatment for 1 day in STZ-diabetic rats. Similar repeated treatment with isoferulic acid reversed the elevated mRNA level of phosphoenolpyruvate carboxykinase (PEPCK) in liver of STZ-diabetic rats to the normal level. However, expression of GLUT4 and PEPCK genes in nondiabetic rats were not influenced by similar treatment with isoferulic acid. These results suggest that isoferulic acid can inhibit hepatic gluconeogenesis and/or increase the glucose utilization in peripheral tissue to lower plasma glucose in diabetic rats lacking insulin. PMID- 10683187 TI - Beta 1-, beta 2- and atypical beta-adrenoceptor-mediated relaxation in rat isolated aorta. AB - beta-adrenoceptor-mediated relaxation was investigated in ring preparations of rat isolated thoracic aorta. Rings were pre-constricted with a sub-maximal concentration of noradrenaline (1 microM) and relaxant responses to cumulative concentrations of beta-adrenoceptor agonists obtained. The concentration-response curve (CRC) to isoprenaline was shifted to the right by propranolol (0.3 microM) with a steepening of the slope. Estimation of the magnitude of the shift from EC(50) values gave a pA(2) of 7.6. Selective beta(1)- and beta(2)-adrenoceptor antagonists, CGP 20712A (0.1 microM) and ICI 118551 (0.1 microM), respectively, produced 4 and 14 fold shifts of the isoprenaline CRC. Atypical beta-adrenoceptor agonists also produced concentration-dependent relaxation of aortic rings. The order of potency of the beta-adrenoceptor agonists was (-log EC(50)): isoprenaline (6. 25)>cyanopindolol (5.59)>isoprenaline+propranolol (5.11)>CGP 12177A (4.40)>ZD 2079 (4.24)>ZM 215001 (4.07)>BRL 37344 (3.89). Relaxation to CGP 12177A and ZM 215001 was unaffected by propranolol (0.3 microM). SR 59230A (O-1184>O-584>O-1315. Log concentration-response curves for O-1238, O-1184 and O-1315 were significantly shifted to the right by SR141716A and the maxima were significantly less than that of the CB(1) agonist, WIN55212-2, an indication of partial agonism. Partial saturation of the triple bond in O-1184 to a cis double bond (O-1238) increased its potency as an agonist (pEC(50) from 6.42 to 7.63) and as an antagonist of WIN55212-2, (pK(B), from 8.36 to 9.49). Substitution of the terminal azide group by an ethyl group (O-584) or removal of the phenolic hydroxyl group (O-1315) had no significant effect on the agonist or antagonist potency. None of these analogues increased the twitch response in a manner resembling that of SR141716A. O-1184 (10 and 100 nM) shifted the log concentration-response curve of WIN55212-2 for inhibition of the twitch responses to the right with pK(B) values of 8.29 and 8.38, respectively. We conclude that these Delta(8)-THC analogues behave as partial agonists rather than silent antagonists at CB(1) binding sites in this tissue. There was no evidence of antagonism of endocannabinoids thus supporting the hypothesis that, in this tissue, SR141716A is an inverse agonist of constitutively active CB(1) receptors. PMID- 10683189 TI - Differential response to chloroethylclonidine in blood vessels of normotensive and spontaneously hypertensive rats: role of alpha 1D- and alpha 1A-adrenoceptors in contraction. AB - The effects of chloroethylclonidine on alpha(1)-adrenoceptor-mediated contraction in endothelium-denuded caudal arteries and aorta from normotensive Wistar and Wistar Kyoto (WKY), and from spontaneously hypertensive (SHR) rats were evaluated. Chloroethylclonidine elicited concentration-dependent contractions. Maximal contraction was similar in caudal arteries among strains ( approximately 40% of noradrenaline effect). However, chloroethylclonidine elicited a higher contraction in aorta from SHR than from normotensive rats. In Wistar aorta chloroethylclonidine produced the smallest contractile response. In SHR aorta, BMY 7378 and 5-methylurapidil blocked chloroethylclonidine-elicited contraction, while (+)-cyclazocine did not inhibit it; while in caudal arteries, 5 methylurapidil blocked chloroethylclonidine action; the other antagonists had no effect. In chloroethylclonidine-treated aorta noradrenaline elicited biphasic contraction-response curves, indicating a high affinity (pD(2), 8.5 - 7.5) chloroethylclonidine-sensitive component and a low affinity (pD(2), 6.3 - 5.2) chloroethylclonidine-insensitive component. The high affinity component was blocked by chloroethylclonidine; while in caudal arteries noradrenaline elicited monophasic contraction-response curves with pD(2) values (6.5 - 5.7) similar to the low affinity component in aorta. Chloroethylclonidine inhibition of noradrenaline response was greater in aorta than in caudal arteries. Chloroethylclonidine increased the EC(50) values of noradrenaline approximately 1000 fold in aorta and approximately 10 fold in caudal arteries. In SHR aorta BMY 7378 protected alpha(1D)-adrenoceptors and in caudal arteries 5-methylurapidil protected alpha(1A)-adrenoceptors from chloroethylclonidine alkylation, allowing noradrenaline to elicit contraction. These results show marked strain-dependent differences in the ability of chloroethylclonidine to contract aorta; moreover, chloroethylclonidine stimulates alpha(1D)-adrenoceptors in aorta and alpha(1A) adrenoceptors in caudal arteries. The higher contraction observed in aorta from SHR and WKY suggests an augmented number of alpha(1D)-adrenoceptors in these strains. PMID- 10683190 TI - Effects of selective inhibitors of cyclo-oxygenase-1 (COX-1) and cyclo-oxygenase 2 (COX-2) on the spontaneous myogenic contractions in the upper urinary tract of the guinea-pig and rat. AB - The role of cyclo-oxygenase-1 (COX-1) and cyclo-oxygenase-2 (COX-2) in the upper urinary tract of the guinea-pig and rat was examined using simultaneous tension recordings of the proximal and distal regions of the renal pelvis and the ureter. The guinea-pig upper urinary tract contracted at a frequency (7.52+/-0.3 min(-1) at 35 degrees C) significantly lower than the frequency in the proximal renal pelvis (21.6+/-1.3 min(-1)) and in the distal renal pelvis and ureter (20.2+/-1.4 min(-1)) of the rat (at 30 degrees C). Indomethacin (>/=1 microM for 60 min), decreased the motility index (amplitudexfrequency) (MI) in all three regions of the guinea-pig upper urinary tract, an effect which mainly arose from a decrease in the frequency of contractions. In the rat, indomethacin (1 - 30 microM for 60 min) significantly decreased the MI calculated in the proximal renal pelvis (>/=30 microM indomethacin), and in the distal renal pelvis (>/=10 microM indomethacin), arising from a significant decrease in the amplitude of contractions. The COX-1 inhibitor, valeryl salicylate (VSA) (5 - 100 microM for 60 min), had no effect on either the amplitude or frequency of contractions in the guinea-pig upper urinary tract. In contrast, VSA increased the force of contractions in the proximal and distal renal pelvis of the rat, whilst having little effect on the frequency of contractions. The COX-2 inhibitor, NS-398 (10 - 100 nM for 60 min) reduced the MI in the guinea-pig upper urinary tract in a concentration-dependent manner. The MIs calculated for the proximal renal pelvis, distal renal pelvis and ureter, were decreased by 72, 64 and 72% respectively, in 100 nM NS-398. NS-398 (10 - 100 nM) had no effect on any of the three parameters measured in either the proximal or distal renal pelvis of the rat. These data suggest that endogenously-released prostaglandins (PGs) maintain the myogenic contractility of the upper urinary tract in both the guinea-pig and rat. Moreover COX-2 is the primary enzyme involved in synthesizing PGs in the guinea-pig upper urinary tract, while COX-1 appears to be the predominantly active enzyme in the rat. PMID- 10683191 TI - The effects of Z13752A, a combined ACE/NEP inhibitor, on responses to coronary artery occlusion; a primary protective role for bradykinin. AB - The effects on the responses to coronary artery occlusion of a combined ACE/NEP inhibitor (Z13752A) were examined in anaesthetized dogs. A 1 h infusion of Z13752A (128 microgram kg(-1) min(-1) intravenously) decreased arterial blood pressure (by 11+/-3%; P<0. 05) and increased coronary blood flow (by 12+/-4%, P<0.05). There were no other significant haemodynamic changes. Z13752A inhibited both NEP and ACE enzymes both in dog plasma and in tissue (lung ACE; kidney NEP). Pressor responses to angiotensin I in vivo were inhibited and systemic vasodilator responses to bradykinin were potentiated. When the left anterior descending coronary artery was occluded for 25 min, Z13752A markedly reduced the severity of the resultant ventricular arrhythmias. No ventricular fibrillation (VF) occurred (compared to 7/16 in the controls; P<0.05), and ventricular tachycardia (VT) was reduced (VT in 2/9 dogs treated with Z13752A cp. 16/16 of controls; episodes of VT 0.2+/-0.1 c.p. 10.7+/-3.3; P<0. 05). Reperfusion of the ischaemic myocardium led to VF in all control dogs but occurred less frequently in dogs given Z13752A (survival from the combined ischaemia-reperfusion insult 67% c.p. 0% in controls; P<0.05). Z13752A reduced two other indices of ischaemia severity; epicardial ST-segment elevation and inhomogeneity of electrical activation. These protective effects of Z13752A during ischaemia and reperfusion were abolished by the administration of icatibant (0.3 mg kg(-1), i.v.) a selective antagonist of bradykinin at B(2) receptors; the ischaemic changes in dogs given both icatibant and Z13752A were similar to those in the controls. We conclude that this ACE/NEP inhibitor is effective at reducing the consequences of coronary artery occlusion in this canine model and that this protection is primarily due to potentiation of released bradykinin. British Journal of Pharmacology (2000) 129, 671 - 680 PMID- 10683192 TI - Wound collagen deposition in rats: effects of an NO-NSAID and a selective COX-2 inhibitor. AB - Selective cyclo-oxygenase (COX)-2 inhibitors and nitric oxide-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) exhibit reduced toxicity in the gastrointestinal tract, but may affect wound healing in other tissues. In this study, we have compared the effects of a selective COX-2 inhibitor (celecoxib), a nitric-oxide releasing derivative of naproxen (HCT-3012) and naproxen in a model of wound collagen deposition in the rat. Polyvinyl alcohol sponges were implanted subcutaneously in rats. The rats were treated daily for 5 days with the test drugs at equieffective anti-inflammatory doses. Naproxen (10 mg kg(-1)) significantly decreased (45%) collagen deposition at the wound site relative to the vehicle-treated control group. In contrast, HCT-3012 (14.5 mg kg(-1)) significantly increased (62%) collagen deposition, while celecoxib (10 mg kg(-1)) had no effect. Naproxen and HCT-3012 suppressed prostaglandin (PG) E(2) levels at the wound site and whole blood thromboxane synthesis to similar degrees. Celecoxib had no significant effect on wound fluid PGE(2) levels, but slightly reduced whole blood thromboxane synthesis (by 17%). COX-1 mRNA and protein were expressed in the wound exudate, the skin surrounding the wound and in normal skin. In contrast, COX-2 mRNA, but not protein, was expressed in wound and normal skin. These results demonstrate that HCT-3012 can significantly enhance collagen deposition at a wound site, despite inhibiting prostaglandin synthesis to the same extent as the parent drug. Nitric oxide-releasing NSAIDs may represent a safer alternative to standard NSAIDs for use as anti-inflammatory and analgesic agents by post-surgery patients. PMID- 10683193 TI - Hamster pancreatic beta cell lines with altered sensitivity towards apoptotic signalling by phosphatase inhibitors. AB - Specific inhibitors of serine/threonine phosphatases like okadaic acid can induce apoptotic cell death in the pancreatic beta cell line HIT. Cultivation in stepwise increased concentrations of okadaic acid enabled the isolation of HIT100R cells which proliferate at 100 nM okadaic acid (8 - 10 times the initially lethal concentration). These two cell lines were used to characterize the events triggered by okadaic acid that led to apoptosis. Biochemical markers, e.g. cytochrome c release from mitochondria and increase of caspase-3-like activity, revealed that induction of apoptosis by 100 nM okadaic acid in parental HIT cells started with the release of cytochrome c. In HIT100R cells 500 nM okadaic acid were necessary to induce alterations comparable to those observed with 100 nM okadaic acid in non-resistant HIT cells. In contrast to okadaic acid, the potency of the structurally different phosphatase inhibitor cantharidic acid to induce cytochrome c release, increase of caspase-3-like activity and DNA fragmentation was comparable in HIT and HIT100R cells. Thus, no cross-resistance between these phosphatase inhibitors seemed to exist. Phosphatase activity in extracts from HIT and HIT100R cells did not differ in its total amount or in its sensitivity for okadaic acid. Since higher concentrations of okadaic acid were needed to induce apoptosis in HIT100R cells, a compromised intracellular accumulation of the toxin appeared likely. Functional and structural analysis revealed that this was achieved by the development of the multidrug resistance phenotype in HIT100R cells. The underlying mechanism appeared to be the enhanced expression of the pgp1 but not the pgp2 gene. PMID- 10683194 TI - Voltage-dependent inhibition of the muscarinic cationic current in guinea-pig ileal cells by SK&F 96365. AB - The effects of SK&F 96365 on cationic current evoked either by activating muscarinic receptors with carbachol or by intracellularly applied GTPgammaS (in the absence of carbachol) were studied using patch-clamp recording techniques in single guinea-pig ileal smooth muscle cells. SK&F 96365 reversibly inhibited the muscarinic receptor cationic current in a concentration-, time- and voltage dependent manner producing concomitant alteration of the steady-state I-V relationship shape which could be explained by assuming that increasing membrane positivity increased the affinity of the blocker. The inhibition was similar for both carbachol- and GTPgammaS-evoked currents suggesting that the cationic channel rather than the muscarinic receptor was the primary site of the SK&F 96365 action. Increased membrane positivity induced additional rapid inhibition of the cationic current by SK&F 96365 which was more slowly relieved during membrane repolarization. Both the inhibition and disinhibition time course could be well fitted by a single exponential function with the time constants decreasing with increasing positivity for the inhibition (e-fold per about 12 mV) and approximately linearly decreasing with increasing negativity for the disinhibition. At a constant SK&F 96365 concentration, the degree of cationic current inhibition was a sigmoidal function of the membrane potential with a potential of half-maximal increase positive to about +30 mV and a slope factor of about -13 mV. Increasing the duration of voltage steps at -80 or at 80 mV, increased the percentage inhibition; the degree of inhibition was almost identical at both potentials providing evidence that the same cationic channel was responsible for the cationic current both at negative and at positive potentials. It is concluded that the distinctive and unique mode of SK&F 96365 action on the muscarinic receptor cationic channel is a valuable tool in future molecular biology studies of this channel. PMID- 10683195 TI - Bromoenol lactone enhances the permeabilization of rat submandibular acinar cells by P2X7 agonists. AB - The permeabilizing effect of P2X(7) agonists was tested in rat submandibular acinar cells using the uptake of ethidium bromide as an index. The uptake of ethidium bromide by acini incubated at 37 degrees C in the presence of 1 mM ATP increased with time and reached after 5 min about 10% of maximal uptake measured in the presence of digitonin. The response to ATP was dose-dependent (half maximal concentration around 40 microM) and it was decreased when the temperature was lowered to 25 degrees C. Benzoyl-ATP reproduced the response to ATP (half maximal concentration around 10 microM). UTP or 2-methylthioATP had no effect. The permeabilization in response to ATP was blocked by oxidized ATP and by magnesium and inhibited by Coomassie blue. ATP increased the activity of a calcium-insensitive phospholipase A(2) (iPLA(2)). Bromoenol lactone (BEL) inhibited the iPLA(2) stimulated by ATP but potentiated the uptake of ethidium bromide in response to the purinergic agonist. From these results it is concluded that the activation of P2X(7) receptors permeabilizes rat submandibular acinar cells. The pore-forming activity of the receptor might be negatively regulated by the concomitant activation of the iPLA(2) by the receptor. PMID- 10683196 TI - M-type K+ currents in rat cultured thoracolumbar sympathetic neurones and their role in uracil nucleotide-evoked noradrenaline release. AB - Cultured sympathetic neurones are depolarized and release noradrenaline in response to extracellular ATP, UDP and UTP. We examined the possibility that, in neurones cultured from rat thoracolumbar sympathetic ganglia, inhibition of the M type potassium current might underlie the effects of UDP and UTP. Reverse transcriptase-polymerase chain reaction indicated that the cultured cells contained mRNA for P2Y(2)-, P2Y(4)- and P2Y(6)-receptors as well as for the KCNQ2 and KCNQ3-subunits which have been suggested to assemble into M-channels. In cultures of neurones taken from newborn as well as from 10 day-old rats, oxotremorine, the M-channel blocker Ba(2+) and UDP all released previously stored [(3)H]-noradrenaline. The neurones possessed M-currents, the kinetic properties of which were similar in neurones from newborn and 9 - 12 day-old rats. UDP, UTP and ATP had no effect on M-currents in neurones prepared from newborn rats. Oxotremorine and Ba(2+) substantially inhibited the current. ATP also had no effect on the M-current in neurones prepared from 9 - 12 day-old rats. Oxotremorine and Ba(2+) again caused marked inhibition. In contrast to cultures from newborn animals, UDP and UTP attenuated the M-current in neurones from 9 - 12 day-old rats; however, the maximal inhibition was less than 30%. The results indicate that inhibition of the M-current is not involved in uracil nucleotide induced transmitter release from rat cultured sympathetic neurones during early development. M-current inhibition may contribute to release at later stages, but only to a minor extent. The mechanism leading to noradrenaline release by UDP and UTP remains unknown. PMID- 10683197 TI - Protein phosphatase-protein kinase interplay modulates alpha 1b-adrenoceptor phosphorylation: effects of okadaic acid. AB - In the present work we studied the effect of protein phosphatase inhibitors on the phosphorylation state and function of alpha(1b)-adrenoceptors. Okadaic acid increased receptor phosphorylation in a time- and concentration-dependent fashion (maximum at 30 min, EC(50) of 30 nM). Other inhibitors of protein phosphatases (calyculin A, tautomycin and cypermethrin) mimicked this effect. Staurosporine and Ro 31-8220, inhibitors of protein kinase C, blocked the effect of okadaic acid on receptor phosphorylation. Neither genistein nor wortmannin altered the effect of okadaic acid. The intense adrenoceptor phosphorylation induced by okadaic acid altered the adrenoceptor-G protein coupling, as evidenced by a small decreased noradrenaline-stimulated [(35)S]GTPgammaS binding. Okadaic acid did not alter the noradrenaline-stimulated increases in intracellular calcium or the production of inositol trisphosphate. Our data indicate that inhibition of protein phosphatases increases the phosphorylation state of alpha(1b) adrenoceptors; this effect seems to involve protein kinase C. In spite of inducing an intense receptor phosphorylation, okadaic acid alters alpha(1b) adrenergic actions to a much lesser extent than the direct activation of protein kinase C by phorbol myristate acetate. PMID- 10683198 TI - The actions of ether, alcohol and alkane general anaesthetics on GABAA and glycine receptors and the effects of TM2 and TM3 mutations. AB - The actions of 13 general anaesthetics (diethyl ether, enflurane, isoflurane, methoxyflurane, sevoflurane, chloral hydrate, trifluoroethanol, tribromoethanol, tert-butanol, chloretone, brometone, trichloroethylene, and alpha-chloralose) were studied on agonist-activated Cl(-) currents at human GABA(A) alpha(2)beta(1), glycine alpha(1), and GABA(C) rho(1) receptors expressed in human embryonic kidney 293 cells. All 13 anaesthetics enhanced responses to submaximal (EC(20)) concentrations of agonist at GABA(A) and glycine receptors, except alpha-chloralose, which did not enhance responses at the glycine alpha(1) receptor. None of the anaesthetics studied potentiated GABA responses at the GABA(C) rho(1) receptor. Potentiation of submaximal agonist currents by the anaesthetics was studied at GABA(A) and glycine receptors harbouring mutations in putative transmembrane domains 2 and 3 within GABA(A) alpha(2), beta(1), or glycine alpha(1) receptor subunits: GABA(A) alpha(2)(S270I)beta(1), alpha(2)(A291W)beta(1), alpha(2)beta(1)(S265I), and alpha(2)beta(1)(M286W); glycine alpha(1)(S267I) and alpha(1)(A288W). For all anaesthetics studied except alpha-chloralose, at least one of the mutations above abolished drug potentiation of agonist responses at GABA(A) and glycine receptors. alpha-Chloralose produced efficacious direct activation of the GABA(A) alpha(2)beta(1) receptor (a 'GABA mimetic' effect). The other 12 anaesthetics produced minimal or no direct activation of GABA(A) and glycine receptors. A non-anaesthetic isomer of alpha chloralose, beta-chloralose, was inactive at GABA(A) and glycine receptors and did not antagonize the actions of alpha-chloralose at GABA(A) receptors. The implications of these findings for the molecular mechanisms of action of general anaesthetics at GABA(A) and glycine receptors are discussed. PMID- 10683199 TI - Cholecystokinin-8 enhances nerve growth factor synthesis and promotes recovery of capsaicin-induced sensory deficit. AB - Alterations of nerve growth factor (NGF) expression have been demonstrated during peripheral nerve disease and the impaired expression or synthesis and transportation of NGF has been correlated with the pathogenesis of several peripheral neuropathies. Since exogenous NGF administration seems to cause undesired side-effects, therapeutical strategies based on the regulation of endogenous synthesis of NGF could prove useful in the clinical treatment of these disorders. The aim of the present study was to analyse the effects of exogenous peripheral administration of the neuropeptide cholecystokinin-8 (CCK-8) on endogenous NGF synthesis, NGF mRNA and distribution of peripheral neuropeptides which are known to be regulated by this neurotrophin. To address these questions we studied the effects of capsaicin (CAPS) before and after the administration of CCK-8 on NGF levels, NGF mRNA expression and localization, and the concentration of substance P (SP) and calcitonin gene-related peptide (CGRP) in peripheral tissue These studies demonstrate that administration of the CCK-8 induces an increase of NGF protein and mRNA in peripheral tissue. NGF level in paw skin of CAPS/CCK-8-treated mice is 3 fold higher than in controls (1241+/-110 pg gr(-1) of tissue wet weight versus 414+/-110 pg gr(-1) of controls) and nearly 6 fold higher than in CAPS-treated mice (1241+/-110 pg gr(-1) versus 248+/-27 pg gr( 1)). The increase of NGF is correlated with the recovery of impaired nocifensive behaviour and with an overexpression of SP and CGRP. The evidence that CCK-8 promotes the recovery of sensory deficits suggests a potential clinical use for this neuropeptide in peripheral neuropathies. PMID- 10683200 TI - Investigation of the interaction between nitric oxide and vasoactive intestinal polypeptide in the guinea-pig gastric fundus. AB - The interaction between nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) was investigated in isolated circular smooth muscle cells and strips of the guinea-pig gastric fundus. VIP induced a concentration-dependent inhibition of carbachol-induced contraction in smooth muscle cells with a maximum at 10(-6) M. The relaxation by 10(-6) M VIP was inhibited for 79.1+/-5.8% (mean+/-s.e. mean) by the NO-synthase (NOS) inhibitor L-N(G)-nitroarginine (L-NOARG; 10(-4) M) in a L-arginine reversible way. Also the inducible NOS (iNOS) selective inhibitor N-(3 (acetaminomethyl)-benzyl)acetamide (1400 W; 10(-6) M) inhibited the VIP-induced relaxation, but its inhibitory effect was not reversed by L-arginine. When cells were incubated with the guanylyl cyclase inhibitor 1H-(1,2,4)oxadiazolo(4,3 a)quinoxalin-1-one (ODQ, 10(-6) M), the protein kinase A-inhibitor (R)-p-cyclic adenosine-3', 5'-monophosphothioate ((R)-p-cAMPS, 10(-6) M) and the glucocorticoid dexamethasone (10(-5) M), the relaxant effect of VIP was decreased by respectively 80.9+/-7.6, 77.0+/-11.6 and 87.1+/-4.5%. In circular smooth muscle strips of the guinea-pig gastric fundus, the VIP (10(-9) - 10(-7) M) induced relaxations were not significantly influenced by 10(-4) M L-NOARG, 10(-6) M 1400 W, 10(-6) M ODQ and 10(-5) M dexamethasone. These results suggest that iNOS, possibly induced by the procedure to prepare the smooth muscle cells, is involved in the relaxant effect of VIP in isolated smooth muscle cells but not in smooth muscle strips of the guinea-pig gastric fundus. This study illustrates the importance of the experimental method when studying the influence of NOS inhibitors on the relaxation induced by VIP in gastrointestinal smooth muscle preparations. PMID- 10683201 TI - Evidence that rat hepatocytes co-express functional P2Y1 and P2Y2 receptors. AB - Previous studies have indicated the expression of multiple P2Y receptors by rat hepatocytes although they have not been identified. Here we show by reverse transcriptase-polymerase chain reaction (RT - PCR) that rat hepatocytes express mRNA encoding all of the four cloned rat P2Y receptors (P2Y(1), P2Y(2), P2Y(4) and P2Y(6)). The effects of UTP have been examined on single aequorin-injected rat hepatocytes. The [Ca(2+)](i) transients induced by UTP were indistinguishable from those induced by ATP in the same cell. The modulatory effects of elevated intracellular cyclic AMP concentration were the same on both UTP- and ATP-induced [Ca(2+)](i) transients. UDP, an agonist at the P2Y(6) receptor, failed to induce transients in hepatocytes, indicating that functional P2Y(6) receptors coupled to increased [Ca(2+)](i) are not expressed. The transients evoked by ADP were more sensitive to inhibition by suramin than those induced by either ATP or UTP. Within an individual cell, the transients induced by ATP and UTP were inhibited by the same concentration of suramin. This sensitivity of ATP and UTP responses to suramin suggests action through P2Y(2) rather than P2Y(4) receptors. Co application of 30 microM pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) caused a decrease in frequency and amplitude of transients induced by ADP. ATP- and UTP-induced transients also displayed a decrease in amplitude in response to addition of PPADS, but this was accompanied by an increase in frequency of transients. In conclusion the data presented here are consistent with the co-expression of P2Y(1) and P2Y(2) receptors by rat hepatocytes. PMID- 10683202 TI - Isolation of the serotoninergic 5-HT4(e) receptor from human heart and comparative analysis of its pharmacological profile in C6-glial and CHO cell lines. AB - RT - PCR technique was used to clone the human 5-HT(4(e)) receptor (h5-HT(4(e))) from heart atrium. We showed that this h5-HT(4(e)) receptor splice variant is restricted to brain and heart atrium. Recombinant h5-HT(4(e)) receptor was stably expressed in CHO and C6-glial cell lines at 347 and 88 fmol mg(-1) protein, respectively. Expression of h5-HT(4(e)) receptors at the cell membrane was confirmed by immunoblotting. The receptor binding profile, determined by competition with [(3)H]-GR113808 of a number of 5-HT(4) ligands, was consistent with that previously reported for other 5-HT(4) receptor isoforms. Surprisingly, we found that the rank order of potencies (EC(50)) of 5-HT(4) agonists obtained from adenylyl cyclase functional assays was inversely correlated to their rank order of affinities (K(i)) obtained from binding assays. Furthermore, EC(50) values for 5-HT, renzapride and cisapride were 2 fold lower in C6-glial cells than in CHO cells. ML10302 and renzapride behaved like partial agonists on the h5 HT(4(e)) receptor. These results are in agreement with the reported low efficacy of the these two compounds on L-type Ca(2+) currents and myocyte contractility in human atrium. A constitutive activity of the h5-HT(4(e)) receptor was observed in CHO cells in the absence of any 5-HT(4) ligand and two 5-HT(4) antagonists, GR113808 and ML10375, behaved as inverse agonists. These data show that the h5 HT(4(e)) receptor has a pharmacological profile which is close to the native h5 HT(4) receptor in human atrium with a functional potency which is dependent on the cellular context in which the receptor is expressed. PMID- 10683203 TI - Non-prostanoid prostacyclin mimetics as neuronal stimulants in the rat: comparison of vagus nerve and NANC innervation of the colon. AB - The spontaneous activity of the rat isolated colon is suppressed by prostacyclin analogues such as cicaprost (IC(50)=4.0 nM). Activation of prostanoid IP(1) receptors located on NANC inhibitory neurones is involved. However, several non prostanoids, which show medium to high IP(1) agonist potency on platelet and vascular preparations, exhibit very weak inhibitory activity on the colon. The aim of the study was to investigate this discrepancy. Firstly, we have demonstrated the very high depolarizing potency of cicaprost on the rat isolated vagus nerve (EC(50)=0.23 nM). Iloprost, taprostene and carbacyclin were 7.9, 66, and 81 fold less potent than cicaprost, indicating the presence of IP(1) as opposed to IP(2)-receptors. Three non-prostanoid prostacyclin mimetics, BMY 45778, BMY 42393 and ONO-1301, although much less potent than cicaprost (195, 990 and 1660 fold respectively), behaved as full agonists on the vagus nerve. On re investigating the rat colon, we found that BMY 45778 (0.1 - 3 microM), BMY 42393 (3 microM) and ONO-1301 (3 microM) behaved as specific IP(1) partial agonists, but their actions required 30 - 60 min to reach steady-state and only slowly reversed on washing. This profile contrasted sharply with the rapid and readily reversible contractions elicited by a related non-prostanoid ONO-AP-324, which is an EP(3)-receptor agonist. The full versus partial agonism of the non-prostanoid prostacyclin mimetics may be explained by the markedly different IP(1) agonist sensitivities of the two rat neuronal preparations. However, the slow kinetics of the non-prostanoids on the NANC system of the colon remain unexplained, and must be taken into account when characterizing neuronal IP-receptors. PMID- 10683204 TI - Chlorotoxin does not inhibit volume-regulated, calcium-activated and cyclic AMP activated chloride channels. AB - It was the aim of this study to look for a high-affinity and selective polypeptide toxin, which could serve as a probe for the volume-regulated anion channel (VRAC) or the calcium-activated chloride channel (CaCC). We have partially purified chlorotoxin, including new and homologous short chain insectotoxins, from the crude venom of Leiurus quinquestriatus quinquestriatus (Lqq) by means of gel filtration chromatography. Material eluting between 280 and 420 min, corresponding to fractions 15-21, was lyophilized and tested on VRAC and CaCC, using the whole-cell patch-clamp technique. We have also tested the commercially available chlorotoxin on VRAC, CaCC, the cystic fibrosis transmembrane conductance regulator (CFTR) and on the glioma specific chloride channel (GCC). VRAC and the correspondent current, I(Cl,swell), was activated in Cultured Pulmonary Artery Endothelial (CPAE) cells by a 25% hypotonic solution. Neither of the fractions 16-21 significantly inhibited I(Cl,swell) (n=4-5). Ca(2+)-activated Cl(-) currents, I(Cl,Ca), activated by loading T84 cells via the patch pipette with 1 microM free Ca(2+), were not inhibited by any of the tested fractions (15-21), (n=2-5). Chlorotoxin (625 nM) did neither effect I(Cl,swell) nor I(Cl,Ca) (n=4-5). The CFTR channel, transiently transfected in COS cells and activated by a cocktail containing IBMX and forskolin, was not affected by 1.2 microM chlorotoxin (n=5). In addition, it did not affect currents through GCC. We conclude that submicromolar concentrations of chlorotoxin do not block volume regulated, Ca(2+)-activated and CFTR chloride channels and that it can not be classified as a general chloride channel toxin. PMID- 10683205 TI - Differential effects of ethanol on glycine uptake mediated by the recombinant GLYT1 and GLYT2 glycine transporters. AB - The effects of ethanol on the function of recombinant glycine transporter 1 (GLYT1) and glycine transporter 2 (GLYT2) have been investigated. GLYT1b and GLYT2a isoforms stably expressed in human embryonic kidney 293 (HEK 293) cells showed a differential behaviour in the presence of ethanol; only the GLYT2a isoform was acutely inhibited. The 'cut-off' (alcohols with four carbons) displayed by the n-alkanols on GLYT2a indicates that a specific binding site for ethanol exists on GLYT2a or on a GLYT2a-interacting protein. The non-competitive inhibition of GLYT2a indicates an allosteric modulation by ethanol of GLYT2a activity. Chronic treatment with ethanol caused differential adaptive responses on the activity and the membrane expression levels of these transporters. The neuronal GLYT2a isoform decreased in activity and surface expression and the mainly glial GLYT1b isoform slightly increased in function and surface density. These changes may be involved in some of the modifications of glycinergic or glutamatergic neurotransmitter systems produced by ethanol intoxication. PMID- 10683207 TI - Agonist binding to G-protein coupled receptors. PMID- 10683206 TI - Apamin-sensitive, non-nitric oxide (NO) endothelium-dependent relaxations to bradykinin in the bovine isolated coronary artery: no role for cytochrome P450 and K+. AB - Since cytochrome P(450)-derived metabolites of arachidonic acid and K(+) have been implicated in endothelium-derived hyperpolarizing factor (EDHF)-dependent responses, the aim of this study was to determine whether such factors contribute to non-nitric oxide (NO), endothelium-dependent relaxation to bradykinin (BK) in bovine isolated coronary artery. In rings of artery contracted with U46619 and treated with indomethacin (3 microM) and N(G)-nitro-L-arginine (L-NOARG; 100 microM), relaxation to BK (0.01 nM-0.3 microM) was blocked by approximately 60% after inhibition of K(+) channels with either high extracellular K(+) (high [K(+)](o); 15 - 67 mM) or apamin (0.3 microM). Ouabain (1 microM), an inhibitor of Na(+)/K(+)-ATPase, decreased the sensitivity to BK without affecting the maximum response. In L-NOARG-treated rings, ouabain had no further effect on the relaxation to BK. An inhibitor of inward-rectifying K(+) channels, Ba(2+) (30 microM), had no effect on relaxations to BK in the absence or presence of either L-NOARG or ouabain. KCl (2.5 - 10 mM) elicited small relaxations ( approximately 20%) that were abolished by nifedipine (0.3 microM) and ouabain. Both the high [K(+)](o)/apamin-sensitive relaxation to BK, and the relaxation to the K(ATP) channel-opener, levcromakalim (0.6 microM), were unaffected by the cytochrome P(450) inhibitor, 7-ethoxyresorufin (10 microM), or by co-treatment with a phospholipase A(2) inhibitor, arachidonyl trifluoromethyl ketone (AACOCF(3); 3 microM) and a diacylglycerol (DAG)-lipase inhibitor, 1, 6-bis (cyclohexyloximinocarbonylamino)-hexane (RHC 80267; 30 microM). The non-NO/high [K(+)](o)-insensitive, approximately 40% relaxation to BK was, however, abolished by these treatments. Therefore, neither cytochrome P(450)-derived metabolites of arachidonic acid nor K(+) appear to mediate the EDHF-like relaxation to BK (i.e the non-NO, high [K(+)](o)/apamin-sensitive component) in bovine coronary arteries. Cytochrome P(450)-derived metabolites may be released at higher BK concentrations to act in parallel with NO and the high [K(+)](o)/apamin-sensitive mechanism. PMID- 10683208 TI - Overview of infection control problems: principles in gastrointestinal endoscopy. AB - The practice of flexible gastrointestinal endoscopy has matured significantly in recent years. Unfortunately, two long-standing problems still exist: the complex physical nature inherent to the endoscopes and accessories, and user compliance with established reprocessing guidelines. Improvements have been made, but newer instruments remain comparatively fragile, expensive, and physically complex, and validated data on reprocessing specific instruments is generally lacking. The practice of flexible gastrointestinal endoscopy today, however, is demonstrably safe and beneficial, provided established practice procedures for reprocessing, with emphasis on instrument cleaning, are followed meticulously in each endoscopy center. PMID- 10683209 TI - Infectious complications associated with gastrointestinal endoscopy. AB - Infectious complications resulting from endoscopy rarely occur. This is probably due as much to the efficiency of the gastrointestinal immune system as to effective endoscope disinfection practices. The low incidence may also represent the difficulty linking infections to endoscopy. Recent reports of probable interpersonal transmission of hepatitis C infection during colonoscopy has heightened this concern. This article examines the documented cases of endoscopy related infections and reviews the risk factors for these infections and details guidelines which have been designed to keep the incidence of these complications low. PMID- 10683210 TI - The microbial flora of the gastrointestinal tract and the cleaning of flexible endoscopes. AB - Technologic advances in the last 30 years have resulted in the development of complex, expensive, and heat sensitive medical instrumentation, including flexible gastrointestinal endoscopes. Because of the design complexity and the region of use, gastrointestinal endoscopes present special challenges to cleaning. If instruments are not properly cleaned the disinfection or sterilization procedure can fail and increase the possibility of infection transmission from one patient to another. Although the cleaning process removes intestinal microflora, the washing process itself may introduce a saprophytic or environmental microbial flora. It has been repeatedly shown that endoscope cleaning, not the disinfection or sterilization procedure, controls the success of the reprocessing procedure. PMID- 10683211 TI - Automatic flexible endoscope reprocessors. AB - Reprocessing medical instruments is a complex and controversial discipline. If all instruments were constructed of materials not damaged by heat, pressure, and moisture, instrument reprocessing would be greatly simplified. As the number of novel and complex instruments entering the market continues to increase, periodic review of the health care facility's instrument reprocessing protocols to ensure their safety and effectiveness is important. This article reviews the advantages and the limitations of automatic flexible endoscope reprocessors. PMID- 10683212 TI - FDA guidelines for endoscope reprocessing. AB - The Food and Drug Administration (FDA) relies on guidance documents, such as voluntary consensus standards and professional practice standards when reviewing the manufacturer's endoscope reprocessing instructions included in the instructions for use manual. The FDA does not perform endoscope reprocessing validation studies. The device manufacturer must certify that the reprocessing instructions included in the user manual have been or will be validated. This article discusses the regulatory review of gastrointestinal endoscopes and endoscope reprocessing instructions required prior to marketing. PMID- 10683213 TI - American Society for Gastrointestinal Endoscopy-Society of Gastroenterology Nurses and Associates Endoscope Reprocessing Guidelines. AB - Adequate endoscope reprocessing is one of the most essential functions in any endoscopy setting. Standards and guidelines for reprocessing endoscopes have been established by the Society of Gastroenterology Nurses and Associates (SGNA) and the American Society for Gastrointestinal Endoscopy (ASGE). These standards pertain to any setting in which gastrointestinal endoscopy is performed. Endoscope reprocessing that meets the established standard of practice helps to ensure a patient-ready instrument for all patients, reduces the risk of disease transmission to practitioners and staff, and helps to prolong the life of the endoscope. PMID- 10683214 TI - Reconciliation of FDA and societal guidelines for endoscope reprocessing. AB - Chemical sterilants are used to high-level disinfect semicritical medical devices such as flexible endoscopes. For the chemosterilant to obtain a high level disinfection claim, The Food and Drug Administration requires demonstration of a 6-log reduction of myobacterial inoculum under worst case conditions (2% horse serum added to test sterilant). This testing requirement has led to label product claims of 45 minutes immersion times at 25 degrees C. Review of the scientific data suggests that at least an 8-log reduction in contamination with thorough instrument cleaning, followed by chemical disinfection for 20 minutes immersion at 20 degrees C will achieve high-level disinfection. PMID- 10683215 TI - Noncompliance with FDA and society guidelines for endoscopic reprocessing: implications for patient care. AB - Flexible endoscopic instruments are very valuable in the diagnosis and treatment of patients with gastrointestinal diseases. Current guidelines for reprocessing these instruments between patient use are appropriate. Rigid adherence to these guidelines, however, will be necessary to reassure governmental authorities, other medical authorities, and the public that the risk of infection from these procedures is minimal. PMID- 10683216 TI - Non-United States guidelines for endoscope reprocessing. AB - A worldwide concern has emerged with regard to endoscope disinfection, and many gastrointestinal endoscopy associations have guidelines for proper disinfection of endoscopes and endoscopic accessories. They are not much different from those of the American Society of Gastrointestinal Endoscopy and include mechanical cleaning as the first and most important step followed by immersion in 2% glutaraldehyde for periods ranging from 5 to 20 minutes. Lack of compliance with these guidelines has been noted, however, in 20 to 70% of centers in Europe, the United Kingdom, Australia, and Asia. Automated disinfectors are being universally recommended to ensure better compliance. Two other improvements are use of peracetic acid as an alternative disinfectant and use of a disposable sheath over the endoscope. PMID- 10683217 TI - Newer technologies for endoscope disinfection: electrolyzed acid water and disposable-component endoscope systems. AB - Novel technologies have been designed to improve or replace more conventional methods of endoscope disinfection. Electrolyzed acid water has the potential to decrease the time, toxicity, and cost of endoscope disinfection. Disposable component endoscope systems have the potential to improve the ease of cleaning and disinfection, or eliminate the need altogether. PMID- 10683218 TI - Overview of the problem: reprocessing versus disposal of endoscopic accessories. AB - The decision to reuse or not reuse endoscopic accessories appears simple on the surface. Further reflection, however, reveals that the decision to purchase reusable accessories or reuse disposable accessories is complex and controversial. This article clarifies key issues surrounding the use of disposable and reusable endoscopic accessories. Discussion includes the advantages and disadvantages of disposable and reusable accessories, the issues surrounding reprocessing of reusable equipment, and the issues surrounding the reuse of disposable accessories. PMID- 10683219 TI - Advantages of disposable endoscopic accessories. AB - Despite the prevailing emphasis on falling reimbursements and cost containment, the use of disposable endoscopic accessories has grown tremendously. They offer simplicity of use, certain sterility, and reduced labor costs in exchange for higher purchase costs per procedure and the burden of waste disposal. Disposable accessories provide greater variety, complexity, and utility. They carry a cost burden that may be acceptable when the devices are difficult to reprocess, when they incorporate features that justify the added cost, or when their unit cost approaches purchase plus reprocessing costs for reusable alternatives, such as for biopsy forceps. Units with small volumes may prefer the ease of disposable accessories independent of relative cost issues, while large high-volume units may need to evaluate cost data more carefully to maintain sustainable practices. PMID- 10683220 TI - Advantages of reusable accessories. AB - Despite scant evidence supporting the use of disposable accessories, these devices have been widely disseminated. Manufacturers and governmental regulators, the most devout proponents of one-time use accessories, have framed the issue in economic terms-parsimonious practitioners reusing disposable accessories at the risk of cross-contamination, mechanical failure and product liability. This simplistic view represents revisionist history and ignores the long tradition of reusing these devices. This article reviews the numerous studies that support the safe and cost effective reuse of disposable and reusable accessories. PMID- 10683221 TI - Methodology of reprocessing reusable accessories. AB - Accessory devices used for gastrointestinal endoscopy are often complex and because they enter sterile body cavities, or contact blood because of invasive procedures, they should be sterile before being used for a patient procedure. The complexity of such reusable accessory devices is reviewed, and the critical aspects of cleaning, sterilization, and quality assurance are discussed. The effectiveness of adequate reprocessing between patient use is critical to prevent transmission of infectious diseases from one patient to another. PMID- 10683222 TI - Methodology of reprocessing one-time use accessories. AB - The current climate of falling reimbursements has prompted evaluation of reusing single use (disposable) endoscopic accessories. A number of different accessories have undergone study of both function and sterility after reuse. These studies, albeit limited in number, do show that selected equipment can be sterilized without impairment of function. Nonetheless, despite the data, widespread adoption of accessory reuse remains hampered by lingering concerns over cross contamination and medical and legal risks. PMID- 10683223 TI - Third-party reprocessing of endoscopic accessories. AB - Third-party reprocessing of medical devices labeled for single use is a safe, FDA regulated practice that helps hospitals reduce costs without compromising patient care. Simply because a device is labeled as single use does not mean it cannot be safely reprocessed. To the contrary, the single use label is chosen by the manufacturer, sometimes for economic gain, as there are no formal FDA regulations or standards to distinguish between reusable and single use devices. The current FDA regulatory framework for third-party reprocessors, which emphasizes compliance with FDA quality assurance requirements, is presently under review, and the agency is in the process of developing a new regulatory scheme for reprocessing. PMID- 10683224 TI - A method for application of samples to matrix-assisted laser desorption ionization time-of-flight targets that enhances peptide detection. AB - Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry has become a fundamental tool for the identification and analysis of peptides and proteins. MALDI-TOF is well suited for the analysis of complex biological mixtures because samples are crystallized onto a solid support that can be washed to remove contaminants and salts prior to laser desorption. A number of approaches for immobilizing samples onto MALDI targets have been put forth. These include the use of different chemical matrices and the immobilization of samples onto different solid supports. In large part though, the preparation of MALDI targets has been an empirical exercise that often requires a unique series of conditions for every sample. Here, a simple method for the application of peptide mixtures onto MALDI targets is put forth. This method differs because peptides are added directly to a sample of nitrocellulose dissolved in acetone, allowing them to interact in solution-phase organic solvent. This solution-phase mixture is then spotted to the MALDI target and evaporated, forming a homogenous solid surface for laser desorption. This procedure is robust, highly sensitive, tolerant to detergents, and easily learned. In our hands, the method provides as much as a 10-fold enhancement to the detection of tryptic peptide fragments derived from in-gel digests. PMID- 10683226 TI - Separation of lipooligosaccharides by linear gradient gel electrophoresis. AB - Lipooligosaccharides (LOSs) are one of the major antigenic and immunogenic components on the outer membrane of mucosal Gram-negative bacteria. These glycolipid antigens are in the M(r) range of 3-7 kDa, and SDS/PAGE has been used as an analytical tool. Although we are able to separate relatively higher M(r) LOS components by mini-PAGE, we encounter difficulties in resolving LOS components below 3.6 kDa present in heterogeneous LOS preparations. In the present study, we selected PID2 LOS consisting of six LOS components of 3.0-5.1 kDa as a model LOS and examined mini-PAGE conditions not only to resolve smaller M(r) LOS components but also to retain resolving capability of higher LOS components. We found that mini-PAGE with stepwise and linear gradient gels (glycine-SDS) resolved smaller M(r) LOS components. Mini-PAGE with linear gradient gels gave the best resolution, and LOS components of 3.0-5.1 kDa were separated as tight and even bands. Because of the resolution, LOS components were stained chemically and immunochemically much better than those on continuous or stepwise gradient gels. Our study also showed that preformed tricine-SDS (TSDS) minigels such as 16.5 and 10-20% (linear gradient) did not resolve PID2 LOS, which indicated that heterogeneous LOS preparations may not be fully analyzed by using these TSDS minigels. By using glycine-SDS linear gradient mini-PAGE, we should be able not only to screen expression of LOSs but also to characterize smaller M(r) LOS components present in heterogeneous LOS preparations whose identities may have been neglected in the past. PMID- 10683225 TI - Tubulin carboxypeptidase assay based on the action of the enzyme on [14C]tyrosinated tubulin bound to nitrocellulose membrane. AB - We have developed a method for the determination of tubulin carboxypeptidase activity which is based on the action of the enzyme on the substrate, [14C]tyrosinated tubulin, previously adsorbed on nitrocellulose membrane. In addition to being two to three times more sensitive than previous carboxypeptidase assays, this method allows the determination of dilute enzyme preparations even containing high salt (inhibitory) concentrations. This is a valuable property specially under circumstances in which numerous high salt containing fractions with scarce activity should be analyzed (for example after certain chromatographic stages during enzyme purification). Our method is simpler, less time-consuming, and suitable for multiple, simultaneous determinations and the substrate bound to nitrocellulose can be stored for several months without significant alteration of its properties. Peptidases other than tubulin carboxypeptidase can act on [14C]tyrosinated tubulin bound to nitrocellulose, solubilizing radioactive compounds, suggesting the eventual applicability of this method to assay proteases in general. Other features and advantages of the assay as well as its limitations are discussed. PMID- 10683227 TI - Characterization of ricin heterogeneity by electrospray mass spectrometry, capillary electrophoresis, and resonant mirror. AB - Electrospray mass spectrometry (ES/MS), capillary-zone electrophoresis (CZE), capillary isoelectric focusing (CIEF), and multianalyte resonant mirror are used to evaluate the heterogeneity of samples of ricin toxins extracted from five horticultural varieties of Ricinus communis seeds: R. communis zanzibariensis, carmencita, impala, sanguineus, and gibsonii. The investigation is also extended to the geographical provenance of the beans. Combining mass spectrometry, CE techniques, and resonant mirror results in a powerful analytical tool capable to characterize and differentiate between different varieties of ricin toxins. Each technique complements the others, adding another level of information. This study reveals a large extent of heterogeneity for each cultivar, demonstrating that ricin toxins consist of a series of glycosylated proteins most likely originating from a multigene family. By combining these techniques, it is possible to differentiate between zanzibariensis and the other four varieties, and that variations in the functional characteristics may be observed between the different cultivars. This study demonstrates that knowledge of the variety of R. communis beans used and their geographical provenance is essential before any type of investigation of ricin toxins is carried out. Consequently, any unusual behavior observed can only be attributed to that particular cultivar studied and not automatically extended to include all R. communis varieties. PMID- 10683228 TI - High-performance capillary electrophoresis of sialylated oligosaccharides of human milk. AB - Oligosaccharides in human milk inhibit enteric pathogens in vitro and in vivo. Neutral milk oligosaccharides vary among individuals and over the course of lactation. To study such variation in the acidic milk oligosaccharides, a sensitive, convenient, quantitative method is needed. High-performance capillary electrophoresis of underivatized acidic oligosaccharides with detection by UV absorbance at 205 nm proved to be sensitive to the femtomole level. Eleven standard oligosaccharides ranging from tri- to nonasaccharide (3'-sialyllactose, 6'-sialyllactose, 3'-sialyllactosamine, 6'-sialyllactosamine, disialyltetraose, 3'-sialyl-3-fucosyllactose, sialyllacto-N-tetraose-a, sialyllacto-N-tetraose-b, sialyllacto-N-neotetraose-c, disialyllacto-N-tetraose, and disialomonofucosyllacto-N-neohexaose) were resolved; baseline resolutions of 3' sialyllactose, 6'-sialyllactose, and other structural isomers were achieved. Peak areas were linear from 30 to 2000 pg and were reproducible with a coefficient of variation between 4 and 9%. There was no evidence of quantitative interference of one oligosaccharide with another. In studies using pooled human milk, addition of increasing amounts of authentic standard oligosaccharides produced the expected positive increments in detected values, indicating quantitative recovery without interference by other milk components. The identities of the major sialylated acidic oligosaccharides of pooled human milk agreed with the results of previous studies employing other analytical methods. Comparison of oligosaccharide profiles of milk samples from different donors revealed extensive variation, especially in the structural isomers of sialyllacto-N-tetraose. This sensitive, highly reproducible method requires only simple sample workup and is useful in defining variations in human milk acidic oligosaccharides and investigating their possible relationship with diseases of infants. PMID- 10683229 TI - Development of an offline noncompetitive flow immunoassay for the determination of interleukin-8 in cell samples. AB - A noncompetitive flow immunoassay system (FIA) for the analysis of interleukin-8 (IL-8) in cell samples was developed. Affinity interaction assays based on offline incubation of excess labeled antibodies and antigen (IL-8) were carried out. The residual unbound labeled antibody was trapped in an immunoaffinity column with immobilized IL-8 while the immunocomplex, labeled antibody/IL-8, was detected by a fluorescence detector. Two fluorophores, FLUOS and Cy5.5, were conjugated with IL-8 antibody. Optimization and comparison between the two fluorescent labeled antibodies were performed with regard to pH, antibody concentration, flow rate, injection volume, and association time. Additionally, a horseradish peroxidase enzyme label was used for the conjugation to the anti-IL 8. The enzyme substrate reaction was optimized with respect to temperature and length of the substrate reaction coil. The detection limits were found to be 200 amol using the FLUOS-labeled anti-IL-8 and 1 fmol using the Cy5.5 fluorescence label. The developed FIA technique was applied for the analysis of IL-8 in cell samples. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to identify IL-8 in the cell samples. PMID- 10683230 TI - Evaluation of methods for the determination of mitochondrial respiratory chain enzyme activities in human skeletal muscle samples. AB - The quantification of mitochondrial enzyme activities in skeletal muscle samples of patients suspected of having mitochondrial myopathies is problematic. Therefore, we have evaluated different methods for the determination of activities cytochrome c oxidase and NADH:CoQ oxidoreductase in human skeletal muscle samples. The measurement of cytochrome c oxidase activity in the presence of 200 microM ferrocytochrome c and the detection of NADH:CoQ oxidoreductase as rotenone-sensitive NADH:CoQ(1) reductase resulted in comparable citrate synthase normalized respiratory chain enzyme activities of both isolated mitochondria and homogenates from control human skeletal muscle samples. These methods allowed the precise detection of deficiencies of respiratory chain enzymes in skeletal muscle of two patients harboring only 20 and 27% of deleted mitochondrial DNA, respectively. Therefore, citrate synthase-normalized respiratory chain activities can serve as stable reference values for the determination of a putative mitochondrial defect in human skeletal muscle. PMID- 10683231 TI - Kinetic analysis of the mass transport limited interaction between the tyrosine kinase lck SH2 domain and a phosphorylated peptide studied by a new cuvette-based surface plasmon resonance instrument. AB - We explored the use of a newly developed cuvette-based surface plasmon resonance (SPR) instrument (IBIS) to study peptide-protein interactions. We studied the interaction between the SH2 domain of lck and a phosphotyrosine peptide EPQY*EEIPIYL which was immobilized on a sensor chip. No indications for mass transport limitation (MTL) were observed when standard kinetic approaches were used. However, addition of competing peptide during dissociation revealed a high extent of rebinding. A dissociation rate constant (k(d)) of 0.6+/-0.1 s(-1) was obtained in the presence of large amounts of peptide. A simple bimolecular binding model, applying second-order kinetics for the cuvette system, could not adequately describe the data. Fits were improved upon including a step in the model which describes diffusion of the SH2 domain from the bulk to the sensor, especially for a surface with high binding capacity. From experiments in glycerol containing buffers, it appeared that the diffusion rate decreased with higher viscosity. It is demonstrated that MTL during association and dissociation can be described by the same diffusion rate. A binding constant (K(D)) of 5.9+/-0.8 nM was obtained from the SPR equilibrium signals by fitting to a Langmuir binding isotherm, with correction for loss of free analyte due to binding. An association rate constant k(a) of 1.1(+/-0.2)x10(8) M(-1) x s(-1) was obtained from k(d)/K(D). The values for k(a) and k(d) obtained in this way were 2-3 orders larger than that from standard kinetic analysis, ignoring MTL. We conclude that in a cuvette the extent of MTL is comparable to that in a flow system. PMID- 10683232 TI - Kinetic analysis of the interaction between HIV-1 protease and inhibitors using optical biosensor technology. AB - The interaction between HIV-1 protease and reversible inhibitors was studied by surface plasmon resonance biosensor technology. The steady-state binding level and the time course of association and dissociation could be observed by measuring the binding of inhibitors injected in a continuous flow of buffer to the immobilized enzyme. Fourteen low molecular weight inhibitors (500-700 Da), including the four clinically used HIV-1 protease inhibitors (indinavir, nelfinavir, ritonavir, and saquinavir), were analyzed. Affinities were estimated as B(50) values from a series of sensorgrams at different concentrations of inhibitors. These values were found to be correlated with inhibition constants (K(i)) determined by an enzyme inhibition assay (r(2) = 0.84, logarithmic values). Dissociation rates were estimated at a single saturating concentration of the inhibitors as t(1/2,obs), but these values did not correlate with K(i) (r(2) = 0.26, logarithmic values). Indinavir had the highest affinity (B(50) = 11 nM) and the fastest dissociation (t(1/2,obs) = 500 s) among the clinically used inhibitors while saquinavir had a lower affinity (B(50) = 25 nM) and the slowest dissociation rate (t(1/2,obs) = 6500 s). Since these two inhibitors have similar K(i) values, the differences in dissociation rates reveal important characteristics in the interaction that cannot be obtained by the inhibition studies. The biosensor data are expected to be of greater in vivo relevance since the experiments were performed in a buffer more similar to physiological conditions. PMID- 10683233 TI - Separation of Galfbeta1-->XGlcNAc and Galpbeta1-->XGlcNAc (X = 3, 4, and 6) as the alditols by high-pH anion-exchange chromatography and thin-layer chromatography: characterization of mucins from Trypanosoma cruzi. AB - The O-linked N-acetylglucosamine oligosaccharides in the mucins of Trypanosoma cruzi may contain galactofuranose or galactopyranose, depending on the strain, one of the components being the disaccharide. Since galactofuranose is a site for antibody recognition, it is desirable to have a sensitive method for the detection of the galactofuranosyl structures. In this paper, we present procedures for the separation of Galfbeta1-->XGlcNAc and Galpbeta1-->XGlcNAc (X = 3, 4, and 6) as the corresponding alditols by high-pH anion-exchange chromatography with pulse amperometric detection. All the isomeric disaccharides could be resolved on a CarboPac PA-10 column, the galactofuranose-containing disaccharides being more retained in the column. GlcNAcol and Galfbeta1- >4(Galpbeta1-->6)GlcNAcol could be analyzed in the same run. The compounds could also be separated by thin-layer chromatography on silica gel 60, a convenient method for analysis of the radiolabeled alditols obtained by reductive beta elimination in the presence of NaB(3)H(4). Both methods were applied for the analysis of the O-linked sugars in the mucins of T. cruzi CL 14 and revealed that they contained only N-acetylglucosamine and the disaccharide Galpbeta1-->4GlcNAc. PMID- 10683234 TI - A spectrophotometric method for assay of tannase using rhodanine. AB - A method for assay of microbial tannase (tannin acyl hydrolase) based on the formation of chromogen between gallic acid and rhodanine is reported. Unlike the previous protocols, this method is sensitive up to gallic acid concentration of 5 nmol and has a precision of 1.7% (relative standard deviation). The assay is complete in a short time, very convenient, and reproducible. PMID- 10683235 TI - Resolution of glycoproteins by a lectin gel-shift assay. AB - Gel-shift assays previously described in the literature are based on protein protein or protein-DNA interactions. We show that carbohydrate-lectin interactions can be successfully used to alter the electrophoretic mobility of glycosylated, but not nonglycosylated, protein species in SDS-polyacrylamide gels. We were able to separate the two closely migrating mono- (95 kDa) and nonglycosylated (92 kDa) forms of a polytopic membrane protein, anion exchanger 1 (AE1), synthesized by cell-free translation or in transfected HEK293 cells. Concanavalin A was selected as the lectin due to the high mannose content of the oligosaccharide chain on AE1. Concanavalin A was either added to the samples prior to loading or copolymerized in a top layer of the separating gel, the latter being the method of choice. The presence of concanavalin A resulted in slower mobility of the monoglycosylated protein while the mobility of the nonglycosylated form was not altered. The shift in mobility was dependent on concentration of concanavalin A and the length of separating gel containing copolymerized concanavalin A. When a diglycosylated mutant of AE1 was tested, good separation was achieved at lower concentrations of concanavalin A. This lectin gel-shift assay allows the separation of N-glycosylated and nonglycosylated forms of the protein. PMID- 10683236 TI - A complete system for identifying inhibitors of creatine kinase B. AB - We have developed a complete system for discovery of lead compounds as inhibitors of creatine kinase B. In this article, we describe production and purification of the recombinant protein, conditions and features of an optimized high-throughput screening assay, and results of our implementation of the system using a diverse compound library. PMID- 10683237 TI - Pulsed high-field gradient in vivo NMR spectroscopy to measure diffusional water permeability in Corynebacterium glutamicum. AB - Pulsed high-field gradient in vivo NMR spectroscopy was used to measure diffusional water permeability in cell suspensions of the Gram-positive bacterium Corynebacterium glutamicum. Two different regions of H2O mobility were detected. One was characterized by the apparent coefficient of self-diffusion, D(1 app) = (4.6-12.7)x10(-8) cm(2) s(-1), depending on the observation time t. The other region was characterized by D(2) = 1.4x10(-5) cm(2) s(-1). The value of D(2) was similar to the diffusion coefficient of H2O in free water and in extracellular biological fluids. Restricted diffusion could be demonstrated for the slower process (D(1)). It was attributed to the cytoplasm of the cells. The membrane permeability, P(d H2O), for C. glutamicum was (4.8+/-0.4)x10(-3) cm s(-1). It compared favorably with values reported for human erythrocytes and was higher by a factor of about 100 compared to the diffusional permeability for ethanol, P(d ethanol), in Zymomonas mobilis. Addition of HgCl2, a water channel inhibitor in eukaryotes, decreased P(d H2O) in C. glutamicum by a factor of approximately 8. To our knowledge, these are the first functional studies of water transport in prokaryotes that yielded quantitative data, viz., transmembrane water permeability expressed through D(H2O) and P(d H2O). PMID- 10683238 TI - Determination of pineal melatonin by precolumn derivatization reversed-phase high performance liquid chromatography and its application to the study of circadian rhythm in rats and mice. AB - Determination of minute amounts of endogenous melatonin in rat and mouse pineal gland was performed using an RP-HPLC system. Melatonin was separated following precolumn derivatization and determined with a fluorescence detector at the emission wavelength of 380 nm with the excitation at 245 nm. The calibration curve of melatonin constructed by adding known amounts of melatonin to the homogenates of mouse pineal gland was linear over the range of 1-500 fmol (injection amount/20 microl). The detection limit of added melatonin was 1 fmol (S/N = 5). Repeatability and day-to-day precision for the melatonin spiked sample of mouse pineal gland was 4.0 and 3.8% (RSD), respectively. Using the present method, circadian changes of melatonin content in rat (Wistar) and mouse (C3H) pineal gland were determined. In addition, a minute amount of melatonin in ddY mouse pineal gland was determined, because pineal melatonin of many inbred mouse strains has been reported to be lower than the detection limit. PMID- 10683239 TI - Optimal use of the fluorescent PicoGreen dye for quantitative analysis of amplified polymerase chain reaction products on microplate. PMID- 10683240 TI - Evaluation of subtilisin cleavage specificity for RNase A peptide bonds by high performance liquid chromatography and mass spectrometric analysis. PMID- 10683241 TI - Degenerate oligonucleotide-primed preamplification of ancient DNA allows the retrieval of authentic DNA sequences. PMID- 10683242 TI - Alkaline treatment after X-Gal staining reaction for Escherichia coli beta galactosidase enhances sensitivity. PMID- 10683243 TI - Decreased expression of adenosine kinase in streptozotocin-induced diabetes mellitus rats. AB - Adenosine has been implicated as an important endogenous regulator of various tissue functions. In diabetes, the responsiveness of several tissues to adenosine is altered. The aim of this study was to investigate the activities of enzymes metabolizing adenosine in tissues of diabetic rats. The cytosolic activity (V(max)) of adenosine kinase (AK) was decreased by 50% in the kidney and by 40% in the heart and liver of diabetic rats. A decrease in the V(max) of AK in diabetic tissues was not associated with a change in the K(m) for adenosine. Evaluation of AK gene transcript status showed significantly lower levels of AK mRNA in diabetic tissues as compared to normal tissues. In diabetic kidneys, the level of AK gene transcript was lowered by 50% on first day after streptozotocin administration, and these reduced levels were sustained declined during the next 10 days. Smaller changes in AK gene transcript levels were observed in the heart and liver than in the kidney. The cytosolic activities of 5'-nucleotidase, AMP deaminase, and adenosine deaminase were unchanged in kidney, heart, and liver of diabetic rats. These results suggest that the turnover of the AMP-adenosine metabolic cycle might be impaired in diabetic tissues due to the reduced activity of adenosine kinase. PMID- 10683244 TI - Cystic fibrosis transmembrane conductance regulator: the purified NBF1+R protein interacts with the purified NBF2 domain to form a stable NBF1+R/NBF2 complex while inducing a conformational change transmitted to the C-terminal region. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is known to function as a regulated chloride channel and, when genetically impaired, to cause the disease cystic fibrosis. The novel studies reported here were undertaken to gain greater molecular insight into possible interactions among CFTR's soluble domains, which include two nucleotide binding domains (NBF1 and NBF2) and a regulatory domain (R). The NBF1+R and NBF2 regions of CFTR were highly expressed in Escherichia coli, purified to near homogeneity under denaturing conditions, and refolded. Both refolded proteins bound TNP-ATP and TNP-ADP, which could be readily replaced with ATP. Four different approaches were then used to determine whether the NBF1+R and NBF2 proteins interact. First, the purified NBF2 protein was labeled near its C-terminus with a fluorescent probe, 7-diethyl amino-3-(4' maleimidylphenyl)-4-methylcoumarin (CPM). Addition of the unlabeled NBF1+R to the CPM-labeled NBF2 caused a red-shift in lambda(max) of the CPM fluorescence, consistent with a direct interaction between the two proteins. Second, when the NBF1+R protein, the NBF2 protein, and a mixture of the two proteins were folded separately and analyzed by molecular sieve chomatography, the mixture was found to elute prior to either NBF1+R or NBF2. Third, na-tive-PAGE gel studies revealed that the mixture of the NBF1+R and NBF2 domains migrated as a single band with an R(F) value between that of NBF1+R and NBF2. Fourth, trypsin digestion of a mixture of the NBF1+R and NBF2 proteins occurred at a slower rate than that for the individual proteins. Finally, studies were carried out to determine whether an NBF1+R/NBF2 interaction could be demonstrated after expressing one of the two proteins in soluble, native form, thus avoiding the inclusion body, denaturation, and renaturation approach. Specifically, the NBF1+R protein was overexpressed in E. coli in fusion with glutathione-S-transferase near a thrombin cleavage site. Following binding of the GST-(NBF1+R) fusion protein to a GST Sepharose affinity column, added NBF2 was shown to bind and then to coelute with NBF1+R upon addition of glutathione or thrombin. Collectively, these experiments demonstrate that CFTR's NBF1+R region and its NBF2 domain, after folding separately as distinct units, have a strong propensity to interact and that this interaction is stable in the absence of added nucleotides or exogenously induced phosphorylation. These findings, together with the additional observation that the NBF1+R/NBF2 interaction induces a change in the C-terminus of NBF2, which resides within the C-terminal region of CFTR, may have important implications not only for the function of CFTR per se, but its interaction with other proteins. PMID- 10683245 TI - Substrate oxidation and ATP supply in AS-30D hepatoma cells. AB - The oxidation of several metabolites in AS-30D tumor cells was determined. Glucose and glycogen consumption and lactic acid production showed high rates, indicating a high glycolytic activity. The utilization of ketone bodies, oxidation of endogenous glutamate, and oxidative phosphorylation were also very active: tumor cells showed a high respiration rate (100 ng atoms oxygen (min x 10(7) cells)(-1)), which was 90% oligomycin-sensitive. AS-30D tumor cells underwent significant intracellular volume changes, which preserved high concentrations of several metabolites. A high O(2) concentration, but a low glucose concentration were found in the cell-free ascites liquid. Glutamine was the oxidizable substrate found at the highest concentration in the ascites liquid. We estimated that cellular ATP was mainly provided by oxidative phosphorylation. These data indicated that AS-30D hepatoma cells had a predominantly oxidative and not a glycolytic type of metabolism. The NADH ubiquinol oxido reductase and the enzyme block for ATP utilization were the sites that exerted most of the control of oxidative phosphorylation (flux control coefficient = 0.3-0.42). PMID- 10683246 TI - Homology models of mu-opioid receptor with organic and inorganic cations at conserved aspartates in the second and third transmembrane domains. AB - Metal ions affect ligand binding to G-protein-coupled receptors by as yet unknown mechanisms. In particular, Na(+) increases the affinity for antagonists but decreases it for agonists. We had modeled the mu-opioid receptor (muR) based on the low-resolution structure of rhodopsin by G. F. X. Schertler, C. Villa, and R. Henderson (1993, Nature 362, 770-772) and proposed that metal ions may be directly involved in the binding of ligands and receptor activation (B. S. Zhorov and V. S. Ananthanarayanan, 1998, J. Biomol. Struct. Dyn. 15, 631-637). Developing this concept further, we present here homology models of muR using as templates the structure of rhodopsin elaborated by I. D. Pogozheva, A. L. Lomize, and H. I. Mosberg (1997, Biophys. J. 70, 1963-1985) and J. M. Baldwin, G. F. X. Schertler, and V. M. Unger (1997, J. Mol. Biol., 272, 144-164). Using the Monte Carlo minimization (MCM) method, we docked the Na(+)-bound forms of muR ligands: naloxone, bremazocine, and carfentanyl. The resultant low-energy complexes showed that the two positive charges in the protonated metal-bound ligands interact with the two negative charges at Asp(3.32) and Asp(2.50) (for notations, see J. A. Ballesteros and H. Weinstein, 1995, Methods Neurosci. 25, 366-426). MCM computation on morphine docked inside the model of muR by I. D. Pogozheva, A. L. Lomize, and H. I. Mosberg (1998, Biophys. J. 75, 612-634) yielded two binding modes with the ligand's ammonium group salt-bridged either to Asp(3.32) (generally regarded as the ligand recognition site) or to Asp(2.50). The latter is the presumed site for Na(+) ion, which is known to modulate ligand binding. Assuming that in the low-dielectric transmembrane region of muR, organic and inorganic cations would compete for Asp(3.32) and Asp(2.50), we propose that ligand binding, as visualized in the above models, would first displace Na(+) from Asp(3.32). A subsequent progress of the ligand toward Asp(2.50) would result in either the retention of Na(+) at Asp(2.50) in the case of antagonists or the displacement of Na(+) from Asp(2.50) in the case of agonists. The displaced Na(+) would move toward the salt-bridged Asp(3.49)-Arg(3.50) and disengage the salt bridge. This, in turn, would result in conformational changes at the cytoplasmic face of the receptor that facilitate the interaction with the G-protein. PMID- 10683247 TI - Hydrogen peroxide-mediated protein oxidation in young and old human MRC-5 fibroblasts. AB - It is suggested that the aging process is dependent on the action of free radicals. One of the highlights of age-related changes of cellular metabolism is the accumulation of oxidized proteins. The present investigation was undertaken to reveal the proliferation-related changes in the protein oxidation and proteasome activity during and after an acute oxidative stress. It could be demonstrated that the activity of the cytosolic proteasomal system declines during proliferative senescence of human MRC-5 fibroblasts and is not able to remove oxidized proteins in old cells efficiently. Whereas in young cells removal of oxidized proteins was accompanied by an increase in the overall protein turnover, this increase in protein turnover could not be seen in old MRC-5 fibroblasts. Therefore, our studies demonstrate that old fibroblasts are much more vulnerable to the accumulation of oxidized proteins after oxidative stress and are not able to remove these oxidized proteins as efficiently as young fibroblasts. PMID- 10683248 TI - Experimental hyperthyroidism causes inactivation of the branched-chain alpha ketoacid dehydrogenase complex in rat liver. AB - Hyperthyroidism induced by 3-day treatment of rats with thyroid hormone (T(3); 3,5,3'-triiodothyronine) at 0.1 or 1 mg/kg body wt/day resulted in a reduced activity state (% of enzyme in its active, dephosphorylated state) of the hepatic branched-chain alpha-ketoacid dehydrogenase (BCKDH) complex. One treatment with 0.1 mg T(3)/kg body wt caused a significant effect on the activity state of BCKDH complex after 24 h, indicating that the reduction of the activity state was triggered by the first administration of T(3). Hyperthyroidism also caused a stable increase in BCKDH kinase activity, the enzyme responsible for phosphorylation and inactivation of the BCKDH complex, suggesting that T(3) caused inactivation of the BCKDH complex by induction of its kinase. Western blot analysis also revealed increased amounts of BCKDH kinase protein in response to hyperthyroidism. No change in the plasma levels of branched-chain alpha-keto acids was observed in T(3)-treated rats, arguing against an involvement of these known regulators of BCKDH kinase activity. Inactivation of the hepatic BCKDH complex as a consequence of overexpression of its kinase may save the essential branched-chain amino acids for protein synthesis during hyperthyroidism. PMID- 10683249 TI - Characterization and partial purification of microsomal NAD(P)H:quinone oxidoreductases. AB - Quinone oxidoreductases are flavoproteins that catalyze two-electron reduction and detoxification of quinones. This leads to the protection of cells against toxicity, mutagenicity, and cancer due to exposure to environmental and synthetic quinones and its precursors. Two cytosolic forms of quinone oxidoreductases [NAD(P)H:quinone oxidoreductase 1 (NQO1) and NRH:quinone oxidoreductase 2 (NQO2)] were previously identified, purified, and cloned. A role of cytosolic NQO1 in protection of cells from oxidative stress, cytotoxicity, and mutagenicity of quinones was established. Currently, we have characterized and partially purified the NQO activity from rat liver microsomes. This activity was designated as microsomal NQO (mNQO). The mNQO activity showed significantly higher affinity for NADH than NADPH as electron donors and catalyzed reduction of 2,6 dichlorophenolindophenol and menadione. The mNQO activity was insensitive to dicoumarol, a potent inhibitor of cytosolic NQO1. Western analysis of microsomal proteins revealed 29- and 18-kDa bands that cross-reacted with polyclonal antibodies raised against cytosolic NQO1. The mNQO activity was partially purified by solubilization of microsomes with detergent Chaps, ammonium sulfate fractionation, and DEAE-Sephacel column chromatography. The microsomal mNQO proteins are expected to provide additional protection after cytosolic NQOs against quinone toxicity and mutagenicity. PMID- 10683250 TI - Sphingosine-1-phosphate activates phospholipase D in human airway epithelial cells via a G protein-coupled receptor. AB - Sphingosine-1-phosphate (SPP) acts as a first messenger in immortalized human airway epithelial cells (CFNPE9o(-)), possibly interacting with an Edg family receptor. Expression of the SPP receptors Edg-1 and Edg-3, as well as a low level of Edg-5/H218, was detected in these cells, in agreement with their ability to specifically bind SPP. The related lipids, lysophosphatidic acid and sphingosylphosphorylcholine, were unable to displace SPP from its high affinity binding sites, suggesting that the biological responses to these different lysolipids are mediated by distinct receptors. SPP markedly inhibited forskolin stimulated cAMP accumulation in a dose-dependent manner and caused a remarkable elevation of intracellular calcium, both effects being sensitive to pertussis toxin treatment. Most importantly, SPP stimulated phosphatidic acid formation, which was maximal after 2 min and decreased within 8-10 min. In the presence of butan-1-ol, suppression of SPP-induced phosphatidic acid formation and production of phosphatidylbutanol were found, clearly indicating activation of phospholipase D (PLD). This finding was also confirmed by analysis of the fatty acid composition of phosphatidic acid, showing an increase in the monounsaturated oleic acid only. The decrease of phosphatidic acid level after 8-10 min incubation with SPP was accompanied by a parallel increase of diacylglycerol production, which was abolished in the presence of butan-1-ol. This result indicates that activation of phospholipase D is followed by stimulation of phosphatidate phosphohydrolase activity. Phosphatidic acid formation was insensitive to protein kinase C inhibitors and almost completely inhibited by pertussis toxin treatment, suggesting that SPP activates phospholipase D via a G(i/o) protein-coupled receptor. PMID- 10683251 TI - A winged helix protein from yeast Saccharomyces cerevisiae recognizes centromere sequences. AB - The winged helix-turn-helix motif was initially identified in the mammalian hepatocyte-enriched transcription factor HNF-3 and the Drosophila forkhead homeotic protein. Proteins containing the winged helix motif have been shown to play important roles in tissue-specific developmental regulation. In this report, by using a genomic binding site selection method, we demonstrate that the winged helix protein YFKH-1 from the yeast Saccharomyces cerevisiae recognizes conserved sequence in yeast centromeres. Thus, our data suggest that the winged helix proteins of the yeast may be involved in centromeric functions of the yeast. PMID- 10683252 TI - Ribosomal stalk protein phosphorylating activities in Saccharomyces cerevisiae. AB - With ribosomal P protein as a substrate, five peaks of protein kinase activity are eluted after chromatography of a Saccharomyces cerevisiae cellular extract on DEAE-cellulose. Two of them correspond to CK-II and the other three have been called RAP-1, RAP-II, and RAP-III. RAP-I was previously characterized. RAP-III is present in a very small amount, which hindered its purification. RAP-II was further purified on phosphocellulose, heparin-Sepharose, and P protein-Sepharose, studied in detail, and compared with other acidic protein kinases, including RAP I, CK-II, and PK60. RAP-II is shown by SDS-PAGE and centrifugation on glycerol linear density gradients to have a molecular mass of around 62 kDa and it is immunologically different from RAP-I and PK60. RAP-II phosphorylates the P proteins in the last serine residue at the highly conserved carboxyl terminal domain as other P-protein kinases. The ribosome-bound stalk P proteins are not equally phosphorylated by the different kinases. Thus, RAP-II and PK60 mainly phosphorylate P1beta and P2alpha whereas RAP-I and CK-II modify all of them. A comparative study of the K(m) and V(max) of the phosphorylation reaction by the different kinases using individual purified acidic proteins suggests changes in the substrate susceptibility upon binding to the ribosome. All the data available reveal clear differences in the characteristics of the various P protein kinases and suggest that the cell may use them to differentially modify the stalk depending, perhaps, on metabolic requirements. PMID- 10683253 TI - Expression of a 70-kDa immunoreactive form of bile salt-dependent lipase by human pancreatic tumoral mia PaCa-2 cells. AB - This work describes the characterization of an immunoreactive form of bile salt dependent lipase (BSDL) expressed by the human pancreatic tumoral Mia PaCa-2 cell line. This BSDL-related protein, which has an M(r) of 70 kDa, is enzymatically active and poorly secreted. Furthermore, a protein with the same electrophoretic migration can also be immunoprecipitated with polyclonal antibodies specific for the human pancreatic BSDL after in vitro translation of RNA isolated from Mia PaCa-2 cells. These data indicated that this BSDL-related protein might be poorly, or not, glycosylated. Reverse transcription and amplification of RNA extracted from Mia PaCa-2 cells using primers able to specifically amplify the full-length mRNA of the human BSDL resulted in a detectable 1.8-kb cDNA product, shorter than that of BSDL (2.2 kb). The sequence of this transcript corresponds to the mRNA sequence that codes for the mature human pancreatic BSDL. However, a deletion of 330 bp is located within the 3'-domain of this cDNA. Therefore data allowed us to conclude that the 70-kDa BSDL-related protein is a 612 amino acid length protein and represents a truncated form of BSDL. The deletion of 110 amino acids occurs in the C-terminal region of the protein, which encompasses 6 tandemly repeated sequences instead of the 16 normally present in the sequence of BSDL. Because feto-acinar pancreatic protein (FAPP), which is the oncofetal counterpart of BSDL, is a C-terminally truncated isoform of BSDL, it is suggested that the 70-kDa BSDL-related protein expressed in MiaPaCa-2 cells could be representative of the protein moiety of FAPP. PMID- 10683254 TI - Development of disulfide peptide mapping and determination of disulfide structure of recombinant human osteoprotegerin chimera produced in Escherichia coli. AB - Recombinant human osteoprotegerin chimera is a 90-kDa protein containing a human IgG Fc domain fused to human osteoprotegerin. The molecule is a dimer linked by two intermolecular disulfide bonds and contains eleven intramolecular disulfide bonds per monomer. A cysteine-rich region in osteoprotegerin contains nine disulfide bridges homologous to the cysteine-rich signature structure of the tumor necrosis factor receptor/nerve growth factor receptor superfamily. In this report, we have developed peptide mapping procedures suitable to generate disulfide-containing peptides for disulfide structure assignment of the fusion molecule. The methods employed included proteolytic digestion using endoproteinases Glu-C and Lys-C in combination followed by LC-MS analyses. Disulfide linkages of peptide fragments containing a single disulfide bond were assigned by sequence analysis via detection of (phenylthiohydantoinyl) cystine and/or by MS analysis. Disulfide bonds of a large, core fragment containing three peptide sequences linked by four disulfides were assigned after generation of smaller disulfide-linked peptides by a secondary thermolysin digestion. Disulfide structures of peptide fragments containing two disulfide bonds were assigned using matrix-assisted laser desorption ionization mass spectrometry with postsource decay. Both the inter- and intramolecular disulfide linkages of the chimeric dimer were confirmed. PMID- 10683255 TI - Isolation and characterization of glycophorin from nucleated (chicken) erythrocytes. AB - A sialoglycoprotein fraction was isolated from chicken erythrocytes by two methods based on the phenol extraction or chloroform/2-propanol extraction of differently prepared erythrocyte membranes. Both preparations gave in SDS-PAGE two major PAS-stained bands (GP2 and GP3), which migrated as 60- and 33-kDa species, respectively, compared to reference proteins, or as 44- and 23-kDa molecules, compared to human glycophorins. Some less abundant slower migrating PAS-stained components, antigenically related to GP2 and GP3, also were detected. No evidence for the presence of antigenically distinct glycoproteins of leukosialin type was obtained. Interconversion in SDS-PAGE, similar carbohydrate composition, and similar antigenic properties of GP2 and GP3 indicated that they are a dimer and monomer, respectively, of the same glycoprotein which shows properties that allow it to be classified as a glycophorin. Lectin binding studies and methylation analysis of beta-elimination products of chicken glycophorin preparation showed the presence of O-glycans and N-glycans. The major O-glycans include sialylated Galbeta1-3GalNAc units and more complex GlcNAc containing chains. Among the N-glycans, there are complex-type biantennary structures with a bisecting GlcNAc residue, accompanied by chains with additional antennas linked to alpha-mannose residues. A characteristic feature of the chicken glycophorin is a relatively high proportion of N-glycans to O-glycans, compared to the glycophorin A from human erythrocytes. PMID- 10683256 TI - Double-base lesions are produced in DNA by free radicals. AB - Evidence has been accumulating at the oligomer level that free radical-initiated DNA damage includes lesions in which two adjacent bases are both modified. Prominent examples are lesions in which a pyrimidine base is degraded to a formamido remnant and an adjacent guanine base is oxidized. An assay has been devised to detect double-base lesions based on the fact that the phosphoester bond 3' to a nuclesoside bearing the formamido lesion is resistant to hydrolysis by nuclease P1. The residual modified dinucleoside monophosphates obtained from a nuclease P1 (plus acid phosphatase) digest of DNA can be (32)P-postlabeled using T4 polynucleotide kinase. Using this assay the formamido single lesion and the formamido-8-oxoguanine double lesion were detected in calf thymus DNA after X irradiation in oxygenated aqueous solution. The lesions were measured in the forms d(P(F)pG) and d(P(F)pG(H)), where P(F) stands for a pyrimidine nucleoside having the base degraded to a formamido remnant and G(H) stands for 8-oxo deoxyguanosine. The yields in calf thymus DNA irradiated 60 Gy were 8.6 and 3.2 pmol/microgram DNA, respectively. PMID- 10683257 TI - P(1),P(4)-Diadenosine 5'-tetraphosphate modulates l-arginine and l-citrulline uptake by bovine aortic endothelial cells. AB - We have previously demonstrated that P(1),P(4)-diadenosine 5'-tetraphosphate (Ap(4)A) interacts with high-affinity and low-affinity binding sites on the bovine aortic endothelial cell (BAEC) surface. In this report we demonstrate that Ap(4)A interaction with the lower affinity site modulates l-arginine (l-Arg) and l-citrulline (l-Cit) uptake by BAEC. Competition uptake studies demonstrate that l-Arg and l-Cit uptake occurs through a common transporter system that is sensitive to Ap(4)A. Evidence is also presented that is consistent with Ap(4)A modulating l-Arg uptake by increasing the affinity of l-Arg for the transporter. PMID- 10683258 TI - Monovalent cation activation in Escherichia coli inosine 5'-monophosphate dehydrogenase. AB - Inosine 5'-monophosphate dehydrogenase (IMPDH) catalyzes the oxidation of inosine 5'-monophosphate (IMP) to xanthosine 5'-monophosphate with the concomitant reduction of NAD to NADH. Escherichia coli IMPDH is activated by K(+), Rb(+), NH(+)(4), and Cs(+). K(+) activation is inhibited by Li(+), Na(+), Ca(2+), and Mg(2+). This inhibition is competitive versus K(+) at high K(+) concentrations, noncompetitive versus IMP, and competitive versus NAD. Thus monovalent cation activation is linked to the NAD site. K(+) increases the rate constant for the pre-steady-state burst of NADH production, possibly by increasing the affinity of NAD. Three mutant IMPDHs have been identified which increase the value of K(m) for K(+): Asp13Ala, Asp50Ala, and Glu469Ala. In contrast to wild type, both Asp13Ala and Glu469Ala are activated by all cations tested. Thus these mutations eliminate cation selectivity. Both Asp13 and Glu469 appear to interact with the K(+) binding site identified in Chinese hamster IMPDH. Like wild-type IMPDH, K(+) activation of Asp50Ala is inhibited by Li(+), Na(+), Ca(2+), and Mg(2+). However, this inhibition is noncompetitive with respect to K(+) and competitive with respect to both IMP and NAD. Asp50 interacts with residues that form a rigid wall in the IMP site; disruption of this wall would be expected to decrease IMP binding, and the defect could propagate to the proposed K(+) site. Alternatively, this mutation could uncover a second monovalent cation binding site. PMID- 10683259 TI - Reactivity of the cysteine residues in the protein splicing active center of the Mycobacterium tuberculosis RecA intein. AB - Protein splicing involves the self-catalyzed excision of an intervening polypeptide segment, an intein, from a precursor protein. The first two steps in the protein splicing process lead to the formation of ester intermediates through nucleophilic attacks by the side chains of cysteine, serine, or threonine residues adjacent to the splice junctions. Since both nucleophilic residues in the Mycobacterium tuberculosis RecA intein are cysteine, their reactivities could be compared by sulfhydryl group titration. This was accomplished by using fusion proteins containing a truncated RecA intein modified by mutation to prevent protein splicing, in which the cysteines at the splice junctions were the only sulfhydryl groups. The ability to undergo hydroxylamine-induced cleavage at the upstream splice junction showed that the modified intein was not impaired in the ability to form ester intermediates. Sulfhydryl titration with iodoacetamide, monitored by quantitating the residual thiols after reaction with a maleimide derivative of biotin, revealed a striking difference in the apparent pK(a) values of the cysteines at the two splice junctions. The apparent pK(a) of the cysteine at the upstream splice junction, which initiates the N-S acyl rearrangement leading to the linear ester intermediate, was approximately 8.2, whereas that of the cysteine residue at the downstream splice junction, which initiates the transesterification reaction converting the linear ester to the branched ester intermediate, was about 5.8. This suggests that the transesterification step is facilitated by an unusually low pK(a) of the attacking thiol group. Comparison of the rates of cleavage of the linear ester intermediates derived from the M. tuberculosis RecA and the Saccharomyces cerevisiae VMA inteins by dithiothreitol and hydroxylamine revealed that the former reacted relatively more slowly with dithiothreitol, suggesting that the RecA intein has diverged in the course of evolution to react preferentially with thiolate anions and thus lacks the basic groups that may facilitate nucleophilic attack by thiols in other inteins. PMID- 10683260 TI - Thyroid hormone and dehydroepiandrosterone permit gluconeogenic hormone responses in hepatocytes. AB - The importance of the sn-glycerol- 3-phosphate (G-3-P) electron transfer shuttle in hormonal regulation of gluconeogenesis was examined in hepatocytes from rats with decreased mitochondrial G-3-P dehydrogenase activity (thyroidectomized) or increased G-3-P dehydrogenase activity [triiodothyronine (T(3)) or dehydroepiandrosterone (DHEA) treated]. Rates of glucose formation from 10 mM lactate, 10 mM pyruvate, or 2.5 mM dihydroxyacetone were somewhat less in hypothyroid cells than in cells from normal rats but gluconeogenic responses to calcium addition and to norepinephrine (NE), glucagon (G), or vasopressin (VP) were similar to the responses observed in cells from normal rats. However, with 2. 5 mM glycerol or 2.5 mM sorbitol, substrates that must be oxidized in the cytosol before conversion to glucose, basal gluconeogenesis was not appreciably altered by hypothyroidism but responses to calcium and to the calcium-mobilizing hormones were abolished. Injecting thyroidectomized rats with T(3) 2 days before preparing the hepatocytes greatly enhanced gluconeogenesis from glyc erol and restored the response to Ca(2+) and gluconeogenic hormones. Feeding dehydroepiandrosterone for 6 days depressed gluconeogenesis from lactate or pyruvate but substantially increased glucose production from glycerol in euthyroid cells and restored responses to Ca(2+) in hypothyroid cells metabolizing glycerol. Euthyroid cells metabolizing glycerol or sorbitol use the G-3-P and malate/aspartate shuttles to oxidize excess NADH generated in the cytosol. The transaminase inhibitor aminooxyacetate (AOA) decreased gluconeogenesis from glycerol 40%, but had little effect on responses to Ca(2+) and NE. However, in hypothyroid cells, with minimal G-3-P dehydrogenase, AOA decreased gluconeogenesis from glycerol more than 90%. Thus, the basal rate of gluconeogenesis from glycerol in the euthyroid cells is only partly dependent on electron transport from cytosol to mitochondria via the malate/aspartate shuttle and almost completely dependent in the hypothyroid state, and the hormone enhancement of the rate in euthyroid cells involves primarily the G-3-P cycle. These data are consistent with Ca(2+) being mobilized by gluconeogenic hormones and G-3-P dehydrogenase being activated by Ca(2+) so as to permit it to transfer reducing equivalents from the cytosol to the mitochondria. PMID- 10683261 TI - The "SUN" family: UTH1, an ageing gene, is also involved in the regulation of mitochondria biogenesis in Saccharomyces cerevisiae. AB - Since it was shown in previous work that NCA3 (one of the four genes of the SUN family) is involved in mitochondrial protein synthesis regulation, the effect of the other members of this gene family was tested. UTH1 (but not SUN4 or SIM1) was also shown to interfere with mitochondria biogenesis. In Deltauth1 cells, cytochromes aa(3), c, and b were lowered by 25 and 15%, respectively. In the double-null mutant Deltauth1Deltanca3, only cytochrome aa(3) was lowered by 50% relative to the wild type. However, the ratio of cellular respiration to cytochrome oxidase was greatly enhanced in the double-null mutant. Measurements on whole lysed cells showed that another mitochondrial enzyme, citrate synthase, was also lowered in Deltauth1 and Deltauth1Deltanca3 whereas hexokinase was not. Electron micrographs showed no difference in global mitochondria content in Deltauth1Deltanca3, but mitochondria appeared less dense to electrons compared to the wild type. Cardiolipin and mtDNA were equivalent in parental and mutant strains. Measurements on isolated mitochondria showed that the cyt aa(3)/cyt b ratio was also lowered in Deltauth1Deltanca3, but the control exerted by the oxidase on the respiratory flux was higher. The activity of other mitochondrial complexes versus oxidase was equivalent in mutants compared to the wild type. These results suggest that the protein equipment could be lowered in mitochondria from strains inactivated for UTH1. PMID- 10683262 TI - Acan125 binding to the SH3 domain of acanthamoeba myosin-IC. AB - The domain organization of Acanthamoeba myosin-I, an oligomodular motor protein, includes a potentially important protein interaction module that is mostly uncharacterized. The Src homology 3, SH3, domain of myosin-I binds Acan125, a protein containing at least two consensus ligand binding domains: C-terminal SH3 binding motifs (PXXP) and N-terminal leucine-rich repeats. We report the first affinities determined for an SH3 domain of any myosin, namely, K(d) = 7 microM for a 21-residue synthetic peptide based on the PXXP domain sequence and K(d) = 0.15 microM for the PXXP domain included in the C-terminus of Acan125. These values are consistent with affinities reported for peptides and proteins that associate with SH3. By deletional analysis we show that only the PXXP domain is required for Acan125 to interact with the SH3 domain of Acanthamoeba myosin-IC (AmyoC(SH3)). The synthetic peptide described above at a concentration near the K(d) for SH3 binding blocked the interaction between native AmyoC and Acan125, mapping the interaction to the PXXP domain of Acan125 and the SH3 domain of myosin-I. These results are consistent with prototypical SH3 binding and suggest that a PXXP module is both necessary and sufficient to interact with an SH3 module of myosin-I. PMID- 10683263 TI - Further analysis of maize C(4) pyruvate,orthophosphate dikinase phosphorylation by its bifunctional regulatory protein using selective substitutions of the regulatory Thr-456 and catalytic His-458 residues. AB - In C(4) plants such as maize, pyruvate,orthophosphate dikinase (PPDK) catalyzes the regeneration of the initial carboxylation substrate during C(4) photosynthesis. The primary catalytic residue, His-458 (maize C(4) PPDK), is involved in the ultimate transfer of the beta-phosphate from ATP to pyruvate. C(4) PPDK activity undergoes light-dark regulation in vivo by reversible phosphorylation of a nearby active-site residue (Thr-456) by a single bifunctional regulatory protein (RP). Using site-directed mutagenesis of maize recombinant C(4) dikinase, we made substitutions at the catalytic His residue (H458N) and at this regulatory target Thr (T456E, T456Y, T456F). Each of these affinity-purified mutant enzymes was assayed for changes in dikinase activity. As expected, substituting His-458 with Asn results in a catalytically incompetent enzyme. Substitutions of the Thr-456 residue with Tyr and Phe reduced activity by about 94 and 99%, respectively. Insertion of Glu at this position completely abolished activity, presumably by the introduction of negative charge proximal to the catalytic His. Furthermore, neither the T456Y nor inactive H458N mutant enzyme was phosphorylated in vitro by RP. The inability of the former to serve as a phosphorylation substrate indicates that RP is functionally a member of the Ser/Thr family of protein kinases rather than a "dual-specificity" Ser-Thr/Tyr kinase, since our previous work showed that RP effectively phosphorylated Ser inserted at position 456. The inability of RP to phosphorylate its native target Thr residue when Asn is substituted for His-458 documents that RP requires the His-P catalytic intermediate form of PPDK as its protein substrate. For these latter studies, synthetic phosphopeptide-directed antibodies specific for the Thr(456)-P form of maize C(4) PPDK were developed and characterized. PMID- 10683264 TI - Removal of the tryptophan 139 side chain in Escherichia coli D-3-phosphoglycerate dehydrogenase produces a dimeric enzyme without cooperative effects. AB - Escherichia coli d-3-phosphoglycerate dehydrogenase (PGDH) is a homotetrameric enzyme whose activity is allosterically regulated by l-serine, the end-product of its metabolic pathway. Previous studies have shown that PGDH displays two modes of cooperative interaction. One is between the l-serine binding sites and the other is between the l-serine binding sites and the active sites. Tryptophan 139 participates in an intersubunit contact near the active site catalytic residues. Site-specific mutagenesis of tryptophan 139 to glycine results in the dissociation of the tetramer to a pair of dimers and in the loss of cooperativity in serine binding and between serine binding and inhibition. The results suggest that the magnitude of inhibition of activity at a particular active site is primarily dependent on serine binding to that subunit but that activity can be modulated in a cooperative manner by interaction with adjacent subunits. The disruption of the nucleotide domain interface in PGDH by mutating Trp-139 suggests the potential for a critical role of this interface in the cooperative allosteric processes in the native tetrameric enzyme. PMID- 10683265 TI - Substrate preferences of caffeic acid/5-hydroxyferulic acid 3/5-O methyltransferases in developing stems of alfalfa (Medicago sativa L.). AB - Caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase (COMT, EC 1.2.1.68) catalyzes at least two reactions in lignin biosynthesis. Of its two supposed substrates in the lignin pathway, COMT from most sources methylates 5 hydroxyferulic acid (5HFA) with two to three times higher activity than caffeic acid (CafA). The ratio of activity for 5HFA compared with CafA increases with the developmental age of alfalfa (Medicago sativa L.) stem internodes, from approximately 1:1 in young (third and fourth) internodes to 2:1 in mature (seventh and eighth) internodes. This observation, together with immunoblot analysis using antiserum raised against recombinant alfalfa COMT, suggests the presence of a different form of COMT, having preference for CafA compared with 5HFA, in young internodes. This apparently new O-methyltransferase (COMT II) was separated from the previously characterized COMT (COMT I) by anion exchange and hydrophobic interaction chromatography. COMT I, but not COMT II, was found in mature internodes. COMT II was not recognized by anti-(COMT I) serum. Furthermore, in addition to substrate preference, COMT II differed from COMT I in native relative molecular mass, pH optimum, and its very low K(m) for CafA. The possible physiological role of COMT II is discussed. PMID- 10683266 TI - Cellular and enzymatic studies of N(omega)-propyl-l-arginine and S-ethyl-N-[4 (trifluoromethyl)phenyl]isothiourea as reversible, slowly dissociating inhibitors selective for the neuronal nitric oxide synthase isoform. AB - N(omega)propyl-l-arginine (NPA) and S-ethyl-N-[4 (trifluoromethyl)phenyl]isothiourea (TFMPITU) inhibit selectively the neuronal nitric oxide (NO) synthase (nNOS) isoform. In the presence of Ca(2+) and calmodulin (CaM), NPA and TFMPITU produce a time- and concentration-dependent suppression of nNOS catalyzed NO formation. This suppression of activity occurs by a first order kinetic process as revealed from linear Kitz-Wilson plots but does not depend on catalytic turnover since it occurs in the absence of NADPH. Following full suppression of NO synthetic activity by either NPA or TFMPITU, NO synthesis can be restored slowly by excess arginine or by dilution, indicating that the effects of these agents are reversible. This behavior is consistent with a dissociation of NPA and TFMPITU from nNOS slowed by a conformational transition produced by Ca(2+) CaM-binding. NPA and TFMPITU bind to nNOS rapidly producing a heme-substrate interaction as revealed by difference spectrophotometry. At physiological conditions (100 microM extracellular arginine), NPA and TFMPITU inhibit Ca(2+)-dependent NO formation by GH(3) pituitary cells with IC(50) values of 19 and 47 microM, respectively, but require millimolar concentrations to inhibit NO formation by cytokine-induced RAW 264.7 murine macrophages. The inhibition of NO formation by these agents in GH(3) cells is rapidly reversible and not due to suppression of cellular arginine uptake. PMID- 10683267 TI - Purification and characterization of the human recombinant histidine-tagged prostaglandin endoperoxide H synthases-1 and -2. AB - We have used in vitro mutagenesis to introduce a six residue histidine sequence (His-tag) near the amino terminal end of the human PGHS-1 and -2 and have expressed these proteins using the baculovirus system. The His-tags are located one and two amino acids beyond the signal peptide cleavage sites of PGHS-1 and PGHS-2, respectively, positions that do not affect their activities or sensitivities to nonsteroidal anti-inflammatory drugs. When expressed in sf-21 cells, the His-tagged enzymes have K(m) values for arachidonate, and IC(50) values for inhibition by nonsteroidal anti-inflammatory drugs that are similar to values reported for the nontagged enzymes. The His-tags allowed for purification of the PGHSs by a simplified protocol involving nickel-affinity and anion exchange FPLC chromatography. The specific activities and recoveries for the purified enzymes were as good or better than those reported previously for purification of the non-tagged PGHS. These baculovirus constructs should provide a convenient source for pharmacologic and biophysical studies that require large scale preparation of human PGHSs. PMID- 10683268 TI - Modulation of glutathione synthetic enzymes by acidic fibroblast growth factor. AB - Increasing evidence suggests that glutathione (GSH) synthesis is a regulated process. Documented increases in gamma-glutamylcysteine synthetase (GCS) occur in response to oxidants, in tumors, on plating cells at a low cell density, and with nerve growth factor stimulation, suggesting that GSH synthesis may be related to the cell growth and transformation. Previously, extracellular acidic fibroblast growth factor (FGF-1) has been demonstrated to cause transformation and aggressive cell growth in murine embryonic fibroblasts. In the present investigation, we sought to determine whether FGF-1, with its growth inducing properties, resulted in the modulation of GSH biosynthetic enzymes, GCS and GSH synthetase. Murine fibroblasts transduced with (hst/KS)FGF-1, a chimeric human FGF-1 gene containing a signal peptide sequence for secretion, displayed elevated gene expression of both heavy and light subunits of GCS. Activity of GSH synthetase was also elevated in these cells compared with control cells. Nonetheless, GSH was decreased in the FGF-1-transduced cells along with high energy phosphates, adenine nucleotides, NADH, and the redox poise. However, GSSG was not elevated in these cells. Fibroblasts stably expressing human immunodeficiency virus type 1 Tat, which induces intrinsic FGF-1 secretion, resulted in similar changes in GCS, GS, and GSH. The results suggest that although increases in the enzymes of GSH synthesis are a common response to growth factors, an increase in GSH content per se is not required for altered cell growth. PMID- 10683269 TI - Effects of spinal cord X-irradiation on the recovery of paraplegic rats. AB - Axonal regrowth is limited in the adult CNS, especially in the spinal cord, one of the major sites of traumatic lesions. Pathophysiological changes occurring after spinal cord injury include complex acute, subacute, and late processes. In this study, we assessed whether X-irradiation interferes with the acute/subacute phases, thereby improving the functional recovery of paraplegic animals. Two days after acute compression of adult rat spinal cords, various doses (0, 2, 5, 10, 20 Gy) of X-rays were administered as one single dose to the compression site. The animals were functionally evaluated over the course of 1 month after injury, using the Tarlov scale and the Rivlin and Tator scale. We also designed a "physiological" scale, including an assessment of urinary function and infection, appropriate for the evaluation of spinal-cord-lesioned animals. Behavioral analysis suggested that the high doses, 20 Gy and, to a lesser extent, 5 and 10 Gy, were toxic, as shown by morbidity rate and "physiological" score. The 2-Gy group showed better motor performances than the lesioned nonirradiated (LNI) animals and the 5- and 20-Gy groups. Motor performance in the 5-, 10-, and 20-Gy groups was poorer than that seen in the LNI group. Gliosis was reduced in the 2 Gy group compared to LNI animals, and there was high levels of gliosis in the highly (>/=5 Gy) irradiated animals. There was a 23% less lesion-induced syringomyelia in the 2-Gy group than in the other groups (LNI and 5-20 Gy). Thus, low doses of X-rays may interfere with the formation of syringomyelia and glial scar, thereby facilitating the recovery of paraplegic animals. These findings suggest that low-dose irradiation of the lesion site, in association with other therapies, is a potentially promising treatment for improving recovery after spinal cord injury. PMID- 10683270 TI - Light and confocal microscopic studies of evolutionary changes in neurofilament proteins following cortical impact injury in the rat. AB - Previous studies have shown that traumatic brain injury (TBI) produces progressive degradation of cytoskeletal proteins including neurofilaments (e.g., neurofilament 68 [NF68] and neurofilament 200 [NF200]) within the first 24 h after injury. Thus, we employed immunofluorescence (light and confocal microscopy) to study the histopathological correlates of progressive neurofilament protein loss observed at 15 min, 3 h, and 24 h following unilateral cortical injury in rats. TBI produced significant alterations in NF68 and NF200 immunolabeling in dendrites and cell bodies at contusion sites ipsilateral to injury, as well as in the noncontused contralateral cortex. Changes in immunolabeling were associated with, but not exclusively restricted to, regions previously shown to contain dark shrunken neurons labeled by hematoxylin and eosin staining, a morphopathological response to injury suggesting impending cell death. Immunofluorescence microscopic studies of neurofilament proteins in the ipsilateral cerebral cortex detected prominent fragmentation of apical dendrites of pyramidal neurons in layers 3-5 and loss of fine dendritic arborization within layer 1. While modest changes were observed 15 min following injury, more pronounced loss of dendritic neurofilament immunofluorescence was detected 3 and 24 h following injury. Confocal microscopy also revealed progressive alterations in NF68 immunoreactivity in dendrites following TBI. While some evidence of structural alterations was observed 15 min following TBI, dendritic breaks were readily detected in confocal micrographs from 3 to 24 h following injury. However, disturbances in axonal NF68 by immunofluorescence microscopy in the corpus callosum were not detected until 24 h after injury. These studies confirmed that derangements in dendritic neurofilament cytoskeletal proteins are not exclusively restricted to sites of impact contusion. Moreover, changes in dendritic cytoskeletal proteins are progressive and not fully expressed within the first 15 min following impact injury. These progressive dendritic disruptions are characterized by disturbances in the morphology of neurofilament proteins, resulting in fragmentation and focal loss of NF68 immunofluorescence within apical dendrites. In contrast, alterations in axonal cytoskeletal proteins are more restricted and delayed with no pronounced changes until 24 h after injury. PMID- 10683271 TI - Tanycytes transplanted into the adult rat spinal cord support the regeneration of lesioned axons. AB - During past years a number of therapeutic strategies have been developed in order to stimulate axonal regeneration after traumatic injuries of the spinal cord. Recently, encouraging data have been obtained by grafting specific glial cells such as Schwann cells or olfactory ensheathing glial cells, known to support the regeneration of peripheral or central axons, respectively. In a recent series of studies, we have shown that tanycytes, a particular glial cell type present in the mediobasal hypothalamus, were able to support the regeneration of a variety of axons innervating this region. The aim of the present study was to determine whether tanycytes could also support the regeneration of lesioned spinal axons. Cultured hypothalamic tanycytes and cortical astrocytes were prelabeled with Fast blue (FB) and grafted into the thoracic spinal cord of adult rats. Three weeks after the transplantation, the animals were fixed and spinal cord sections treated for multiple fluorescence detection of the FB-labeled transplanted cells on the one hand and of various glial and neuronal markers on the other hand. We show here that in all the spinal cords examined, transplanted tanycytes or astrocytes formed large spherical clusters of about 0.5 mm in diameter, located in the mediolateral spinal cord layer. The immunodetection of glial markers showed that transplanted astrocytes exhibited intense immunostaining for both glial fibrillary acidic protein (GFAP) and vimentin (VIM), whereas transplanted tanycytes were intensely immunostained for VIM, but GFAP negative. The immunodetection of axonal markers showed that contrasting with astrocyte transplants, tanycyte transplants were invaded by numerous axonal fibers. These data indicate that tanycyte transplants may represent a useful therapeutic tool for the reparation of the lesioned spinal axons. PMID- 10683272 TI - Upregulation of BDNF mRNA and trkB mRNA in the nigrostriatal system and in the lesion site following unilateral transection of the medial forebrain bundle. AB - We have performed unilateral transection of the medial forebrain bundle (MFB) and studied BDNF mRNA and trkB mRNA levels at different postlesion times in the nigrostriatal system by means of in situ hybridization. BDNF mRNA levels were transiently induced in the substantia nigra pars compacta at 1 day postaxotomy. The disposition of BDNF mRNA expressing cells at this postlesion time in substantia nigra mimicked that of the dopaminergic neurons expressing the mRNA for the dopamine transporter. TrkB mRNA levels remained unaltered in the ventral mesencephalon at the different postlesion times examined-1 to 14 days. In contrast, trkB mRNA levels were significantly induced in the striatum at the longer postlesion time examined-14 days-when all neurodegenerative events are completed. It is becoming apparent that nigral BDNF mRNA levels are anterogradely transported to its target tissue in striatum. However, following axotomy, the lesion site represents a second potential target for BDNF action. Consequently, we also analyzed the pattern of mRNA expression for BDNF and trkB at the lesion site where dopaminergic axons are disconnected. There, we found notable inductions of both BDNF mRNA and trkB mRNA levels at 4 days postaxotomy. BDNF mRNA expressing cells were confined at the site of axotomy, which coincided precisely to that showing induction of trkB mRNA. Altogether, our results anticipate promising trophic roles of BNDF in the injured nigrostriatal system. PMID- 10683273 TI - Robust regeneration of CNS axons through a track depleted of CNS glia. AB - Transected CNS axons do not regenerate spontaneously but may do so if given an appropriate environment through which to grow. Since molecules associated with CNS macroglia are thought to be inhibitory to axon regeneration, we have tested the hypothesis that removing these cell types from an area of brain will leave an environment more permissive for axon regeneration. Adult rats received unilateral knife cuts of the nigrostriatal tract and ethidium bromide (EB) was used to create a lesion devoid of astrocytes, oligodendrocytes, intact myelin sheaths, and NG2 immunoreactive cells from the site of the knife cut to the ipsilateral striatum (a distance of 6 mm). The regenerative response and the EB lesion environment was examined with immunostaining and electron microscopy at different timepoints following surgery. We report that large numbers of dopaminergic nigral axons regenerated for over 4 mm through EB lesions. At 4 days postlesion dopaminergic sprouting was maximal and the axon growth front had reached the striatum, but there was no additional growth into the striatum after 7 days. Regenerating axons did not leave the EB lesion to form terminals in the striatum, there was no recovery of function, and the end of axon growth correlated with increasing glial immunoreactivity around the EB lesion. We conclude that the removal of CNS glia promotes robust axon regeneration but that this becomes limited by the reappearance of nonpermissive CNS glia. These results suggest, first, that control of the glial reaction is likely to be an important feature in brain repair and, second, that reports of axon regeneration must be interpreted with caution since extensive regeneration can occur simply as a result of a major glia-depleting lesion, rather than as the result of some other specific intervention. PMID- 10683274 TI - Establishment and properties of a growth factor-dependent, perpetual neural stem cell line from the human CNS. AB - The ready availability of unlimited quantities of neural stem cells derived from the human brain holds great interest for basic and applied neuroscience, including therapeutic cell replacement and gene transfer following transplantation. We report here the combination of epigenetic and genetic procedures for perpetuating human neural stem cell lines. Thus we tested various culture conditions and genes for those that optimally allow for the continuous, rapid expansion and passaging of human neural stem cells. Among them, v-myc (the p110 gag-myc fusion protein derived from the avian retroviral genome) seems to be the most effective gene; we have also identified a strict requirement for the presence of mitogens (FGF-2 and EGF) in the growth medium, in effect constituting a conditional perpetuality or immortalization. A monoclonal, nestin-positive, human neural stem cell line (HNSC.100) perpetuated in this way divides every 40 h and stops dividing upon mitogen removal, undergoing spontaneous morphological differentiation and upregulating markers of the three fundamental lineages in the CNS (neurons, astrocytes, and oligodendrocytes). HNSC.100 cells therefore retain basic features of epigenetically expanded human neural stem cells. Clonal analysis confirmed the stability, multipotency, and self-renewability of the cell line. Finally, HNSC.100 can be transfected and transduced using a variety of procedures and genes encoding proteins for marking purposes and of therapeutic interest (e.g., human tyrosine hydroxylase I). PMID- 10683275 TI - Experimental gliosarcoma induces chemokine receptor expression in rat brain. AB - Macrophage/microglial infiltration is a characteristic feature of brain tumors. The functional role(s) of these cells is complex and could include both trophic and suppressive effects on tumors. Information has recently emerged about the molecular signals that regulate the accumulation and function of monocytes in pathological disorders. Recent data indicate that the chemokine, monocyte chemoattractant protein-1 (MCP-1), a potent monocyte activating and chemotactic factor, is a primary regulator of the macrophage response in brain tumors. We hypothesized that if MCP-1 regulates macrophage/microglial infiltration, then expression of the specific MCP-1 receptor, CCR2, will be induced in peritumoral tissue and/or within brain tumors. Identification of a specific receptor that is preferentially expressed in brain tumors could be important both in terms of tumor biology and as a potential therapeutic target. We used an established experimental gliosarcoma model, induced by intracranial transplantation of cultured 9L cells into adult rat brain, to test this hypothesis. RT-PCR analysis showed high levels of both MCP-1 and CCR2 mRNA and Western blot analysis demonstrated increased CCR2 protein in tumor extracts. Immunocytochemistry showed CCR2 immunoreactive microglia in peritumoral tissue and, unexpectedly, that intrinsic tumor cells, rather than monocytes, were the predominant source of CCR2. These results demonstrate that CCR2 expression is markedly upregulated in this brain tumor model. PMID- 10683276 TI - Engagement of the scavenger receptor is not responsible for beta-amyloid stimulation of monocytes to a neurocytopathic state. AB - Experiments were performed to determine if scavenger receptors (SRs) play a role in amyloid beta (Abeta) stimulation of peripheral blood monocyte (PBM) neurotoxicity. Results indicate that Abeta does not block binding of the SR ligand DiI-acetylated low density lipoprotein to PBM, nor does another SR ligand, fucoidin, inhibit Abeta-PBM binding. Moreover, neither of three SR ligands alone stimulates neurotoxicity in PBM, nor antagonizes the ability of Abeta to activate PBM to a neurocytopathic state. Such findings suggest that Abeta's action is not dependent upon engagement of the SR ligand binding domain and raise doubts about the role of SR in Abeta neurotoxicity. PMID- 10683277 TI - Traumatic brain injury leads to increased expression of peripheral-type benzodiazepine receptors, neuronal death, and activation of astrocytes and microglia in rat thalamus. AB - In mammalian CNS, the peripheral-type benzodiazepine receptor (PTBR) is localized on the outer mitochondrial membrane within the astrocytes and microglia. PTBR transports cholesterol to the site of neurosteroid biosynthesis. Several neurodegenerative disorders were reported to be associated with increased densities of PTBR. In the present study, we evaluated the changes in the PTBR density and gene expression in the brains of rats as a function of time (6 h to 14 days) after traumatic brain injury (TBI). Sham-operated rats served as control. Between 3 and 14 days after TBI, there was a significant increased in the binding of PTBR antagonist [(3)H]PK11195 (by 106 to 185%, P < 0.01, as assessed by quantitative autoradiography and in vitro filtration binding) and PTBR mRNA expression (by 2- to 3. 4-fold, P < 0.01, as assessed by RT-PCR) in the ipsilateral thalamus. At 14 days after the injury, the neuronal number decreased significantly (by 85 to 90%, P < 0.01) in the ipsilateral thalamus. At the same time point, the ipsilateral thalamus also showed increased numbers of the glial fibrillary acidic protein positive cells (astrocytes, by approximately 3.5-fold) and the ED-1 positive cells (microglia/macrophages, by approximately 36-fold), the two cell types known to be associated with PTBR. Increased PTBR expression following TBI seems to be associated with microglia/macrophages than astrocytes as PTBR density at different periods after TBI correlated better with the number of ED-1 positive cells (r(2) = 0.95) than the GFAP positive cells (r(2) = 0.56). TBI-induced increased PTBR expression is possibly an adaptive response to cellular injury and may play a role in the pathophysiology of TBI. PMID- 10683278 TI - Neuronal subclass-selective loss of pyruvate dehydrogenase immunoreactivity following canine cardiac arrest and resuscitation. AB - Chronic impairment of aerobic energy metabolism accompanies global cerebral ischemia and reperfusion and likely contributes to delayed neuronal cell death. Reperfusion-dependent inhibition of pyruvate dehydrogenase complex (PDHC) enzyme activity has been described and proposed to be at least partially responsible for this metabolic abnormality. This study tested the hypothesis that global cerebral ischemia and reperfusion results in the loss of pyruvate dehydrogenase immunoreactivity and that such loss is associated with selective neuronal vulnerability to transient ischemia. Following 10 min canine cardiac arrest, resuscitation, and 2 or 24 h of restoration of spontaneous circulation, brains were either perfusion fixed for immunohistochemical analyses or biopsy samples were removed for Western immunoblot analyses of PDHC immunoreactivity. A significant decrease in immunoreactivity was observed in frontal cortex homogenates from both 2 and 24 h reperfused animals compared to samples from nonischemic control animals. These results were supported by confocal microscopic immunohistochemical determinations of pyruvate dehydrogenase immunoreactivity in the neuronal cell bodies located within different layers of the frontal cortex. Loss of immunoreactivity was greatest for pyramidal neurons located in layer V compared to neurons in layers IIIc/IV, which correlates with a greater vulnerability of layer V neurons to delayed death caused by transient global cerebral ischemia. PMID- 10683279 TI - Complement component C1q modulates the phagocytosis of Abeta by microglia. AB - Recent studies showing that microglia internalize the amyloid beta-peptide (Abeta) suggest that these cells have the potential for clearing Abeta deposits in Alzheimer's disease, and mechanisms that regulate the removal of Abeta may therefore be of clinical interest. Previous studies from this laboratory showing that C1q enhances phagocytosis of cellular targets by rat microglia prompted the current investigations characterizing the effects of C1q on microglial phagocytosis of Abeta. Microglia were shown to phagocytose Abeta1-42, in agreement with observations of other investigators. Uptake of Abeta1-42 was observed for concentrations of 5-50 microM, and phagocytosis of peptides containing (14)C or fluorescein (FM) labels was not affected by the interaction of microglia with C1q-coated surfaces. However, inclusion of C1q (125 nM-1.4 microM) in solutions of 50 microM Abeta1-42 inhibited the uptake of (14)C-Abeta1 42 and FM-Abeta1-42, suggesting that C1q blocks the interaction of Abeta with microglia. Uptake of Abeta was partially blocked by the scavenger receptor ligands polyinosinic acid and maleylated BSA. Inhibition of Abeta uptake by C1q may contribute to the accumulation of fibrillar, C1q-containing plaques that occurs in parallel with disease progression. These data suggest that mechanisms which interfere with the binding of C1q to Abeta may be of therapeutic value both through inhibition of the inflammatory events resulting from complement activation and via altered access of Abeta sites necessary for ingestion by microglia. PMID- 10683280 TI - Genetic dissection of the signals that induce synaptic reorganization. AB - Synaptic reorganization of mossy fibers following kainic acid (KA) administration has been reported to contribute to the formation of recurrent excitatory circuits, resulting in an epileptogenic state. It is unclear, however, whether KA induced mossy fiber sprouting results from neuronal cell loss or the seizure activity that KA induces. We have recently demonstrated that certain strains of mice are resistant to excitotoxic cell death, yet exhibit seizure activity similar to what has been observed in rodents susceptible to KA. The present study takes advantage of these strain differences to explore the roles of seizure activity vs cell loss in triggering mossy fiber sprouting. In order to understand the relationships between gene induction, cell death, and the sprouting response, we assessed the regulation of two molecules associated with the sprouting response, c-fos and GAP-43, in mice resistant (C57BL/6) and susceptible (FVB/N) to KA-induced cell death. Following administration of KA, increases in c-fos immunoreactivity were observed in both strains, although prolonged induction of c fos was present only in the hippocampal neurons of FVB/N mice. Mossy fiber sprouting following KA administration was also only observed in FVB/N mice, while induction of GAP-43, a marker associated with mossy fiber sprouting, was not observed in either strain. These results indicate that: (i) KA-induced seizure activity alone is insufficient to induce mossy fiber sprouting; (ii) mossy fiber sprouting may be due to the loss of hilar neurons following kainate administration; and (iii) induction of GAP-43 is not a necessary component of the sprouting response that occurs following KA in mice. PMID- 10683281 TI - Internalization of intracerebrally administered porcine galanin (1-29) by a discrete nerve cell population in the hippocampus of the rat. AB - In spite of numerous studies utilizing intraventricular administration of porcine galanin (1-29), little is known about the spread and cellular distribution of exogenous galanin following intraventricular administration. In this study a discrete nerve cell body population with their dendrites became strongly galanin immunoreactive (IR) in the dorsal hippocampus following intraventricular porcine galanin (1.5 nmol/rat). Time course experiments showed that after time intervals of 10 and 20 min, but not at 60 min, scattered small- to medium-sized galanin-IR nerve cell bodies and their dendrites were present in all layers of the dorsal and ventral hippocampus. In double-immunolabeling experiments most of these nerve cells were identified as putative GABA interneurons costoring NPY-IR or somatostatin-IR in some cases. Twenty minutes after intraventricular injection of artificial cerebrospinal fluid (aCSF), only endogenous punctate and coarse galanin-IR terminals were found, but no galanin-IR cell bodies. Intrahippocampal injection of fluorophore-labeled galanin resulted in the appearance of fluorescent nerve cell bodies with the same morphology and localization as in the above experiments. Coadministration of the putative galanin antagonist M35 (0.5 nmol) and galanin (1.5 nmol) resulted in a reduced number of galanin-IR nerve cell bodies in the hippocampus of half of the rats. These findings support the existence of a population of putative hippocampal GABA interneurons with the ability to internalize and concentrate galanin and/or its fragments present in the extracellular fluid, possibly mediated by galanin receptors. PMID- 10683282 TI - Prenatal methotrexate exposure decreases seizure susceptibility in young rats of two strains. AB - Effects of prenatal exposure to methotrexate (MTX) administered in Sprague-Dawley (one 5 mg/kg dose of MTX on gestational day 15; E15) or Wistar (one 5 mg/kg dose of MTX on E14 or E15 or two such doses on E15) pregnant rat dams were studied in developing offspring. Young Sprague-Dawley rats were subjected to rapid kindling on postnatal days (PN) 15 and 16, and to flurothyl seizures on PN 15 and PN 30. Offspring of the Wistar strain were tested in flurothyl on PN 30. In Sprague Dawley rats, prenatal exposure to MTX decreased susceptibility to kindling induced seizures on PN 15 and to flurothyl-induced clonic seizures on PN 30. In Wistar rats, a single dose of MTX on E15 was ineffective, but two doses significantly decreased susceptibility to flurothyl-induced seizures. Additionally, due to a shorter duration of pregnancy in Wistar rats, exposure to a single dose of MTX on E14 also decreased susceptibility to flurothyl seizures. MTX, as folic acid antagonist, interferes with DNA synthesis. However, unlike other treatments that suppress DNA synthesis (such as methylazoxymethanol exposure or X-ray radiation), MTX exposure results in anticonvulsant effects in surviving offspring. The data suggest that not all prenatal impairments of DNA have proconvulsant features postnatally. PMID- 10683283 TI - Brain RNA polymerase and nucleolar structure in perinatal asphyxia of the rat. AB - Ribosomes are integral constitutens of the protein synthesis machinery. Polymerase I (POL I) is located in the nucleolus and transcribes the large ribosomal genes. POL I activity is decreased in ischemia but nothing is known so far on POL I in perinatal asphyxia. We investigated the involvement of POL I in a well-documented model of graded systemic asphyxia at the level of activity, mRNA, protein, and morphology. Caeserean section was performed at the 21st day of gestation. Rat pups still in the uterus horns were immerged in a water bath for asphyctic periods from 5-20 min. Brain was taken for measurement of pH, nuclear POL I activity, and mRNA steady state, and protein levels of RPA40, an essential subunit of POL I and III. Silver staining and transmission electron microscopy with morphometry when appropriate were used to examine the nucleolus. Brain pH and nuclear POL I activity decreased with the length of the asphyctic period while POL-I mRNA and protein levels were unchanged. Accompanying the decrease in brain pH we found significant changes of nucleolar structure in the course of perinatal asphyxia at the light and electron microscopic level. As early as ten min following the asphyctic insult, morphological disintegration of the nucleolus was observed. The changes became more dramatic with longer duration of perinatal asphyxia. We conclude that severe acidosis may be responsible for decreased POL activity and for disintegration of nucleoli in neurons. This condition may lower the ribosome content in neonatal neurons and impair protein synthesis. PMID- 10683284 TI - Sciatic nerve transection in the adult rat: abnormal EMG patterns during locomotion by aberrant innervation of hindleg muscles. AB - The effects of lesions in the sciatic nerve were studied in adult rats. In the left hindleg, a segment 12 mm long was resected from the proximal part of the nerve, before the bifurcation into the peroneal and tibial nerves. This segment in a reversed orientation was used as a nerve graft. EMG patterns in the tibialis anterior and the gastrocnemius muscles at both sides were recorded during locomotion in six rats after recovery periods varying from 15 to 21 weeks. The specificity of axonal outgrowth was studied in nine rats by retrogradely labeling the motoneurons with unconjugated Cholera Toxin subunit B (CTB) after injections into the gastrocnemius, the soleus, and the tibialis anterior muscles at both sides. EMG patterns at the operated side were irregular and we often observed coactivation of the gastrocnemius and tibialis anterior muscle. Moreover, burst activity was badly adjusted to the phases of the stepcycle. Retrogradely labeling indicated that the pools of motoneurons innervating the respective muscles at the left side had increased in volume. Neuronal diameters were slightly decreased but a considerable decrease was observed in dendritic branching and dendrite bundles in the pools of the SOL and in the GC were absent. No consistent trends in neuronal numbers at the affected side in comparison to the right side were detected. We conclude that axons, sprouting from the proximal stump of the sciatic nerve, innervate the muscles aselectively and that the motoneurons of origin maintain their original activation pattern. PMID- 10683285 TI - Safety and tolerability assessment of intrastriatal neural allografts in five patients with Huntington's disease. AB - This study describes issues related to the safety and tolerability of fetal striatal neural allografts as assessed in five patients with Huntington's disease. Huntington's disease (HD) is characterized by motor, cognitive, and behavioral disturbances. The latter include psychological disturbances and, as a consequence, we took particular care to analyze behavioral changes, in addition to the usual "safety" follow-up. We conducted multidisciplinary follow-up at least 2 years before and 1 year after grafting. Psychological care extended to close relatives. The grafting procedure itself was altogether safe and uneventful, and there were no apparent clinical deleterious effects for 1 year. The immunosuppressive treatment, however, was complicated by various problems (irregular compliance, errors of handling, side effects). Direct psychological consequences of the transplantation procedure were rare and not worrisome, although mood alteration requiring treatment was observed in one patient. Indirectly, however, the procedure required patients and relatives to accept constraints that tended to complicate familial situations already marred by aggressivity and depression. All patients and close relatives expressed major expectations, in spite of our strong and repeated cautioning. It is clearly important to be aware of these particular conditions since they may eventually translate into psychological difficulties in coping with the long-term clinical outcome of the procedure, if not beneficial. Despite an overall good tolerance, therefore, this follow-up calls for caution regarding the involvement of HD patients in experimental surgical protocols. PMID- 10683286 TI - Reversible physiological alterations in sympathetic neurons deprived of NGF but protected from apoptosis by caspase inhibition or Bax deletion. AB - Cell death in nervous system development and in many neurodegenerative diseases appears to be apoptotic or programmed. Withdrawal of nerve growth factor (NGF) from cultures of superior cervical ganglia neurons (SCG) is an excellent model of programmed cell death (PCD), producing apoptosis within 24-48 h. This death can be prevented by treatment with caspase inhibitors or deletion of the proapoptotic Bax gene. Since inhibition of apoptosis is an attractive strategy for the therapy of many neurological diseases and little is known about the function of neurons when apoptosis has been aborted, we examined the electrophysiological properties of NGF-deprived SCG neurons from rats and mice, saved by the caspase inhibitor boc-aspartyl(OMe)fluoromethyl ketone (BAF) or by Bax deletion. Compared to NGF maintained controls, the resting membrane potentials of BAF-saved neurons were depolarized by 9 mV and the action potentials were prolonged by over 50%. Nicotinic cholinergic current density was depressed by about 50%. Electrophysiological parameters returned to normal within 4 days after NGF restoration. Neurons from Bax-deficient mice were altered differently by NGF withdrawal. There were no detectable changes in resting or action potentials. However, nicotinic current density was reduced just as in BAF-saved rat neurons. There were no observable changes in the processes of individual neurons after 6 days of NGF deprivation in the presence of BAF. Our results indicate that neurons are physiologically altered during pharmacological inhibition of PCD, but fully recover after trophic support is returned. PMID- 10683287 TI - Effect of acute L-Dopa pretreatment on apomorphine-induced rotational behavior in a rat model of Parkinson's disease. AB - Currently, reduction of apomorphine-induced rotational behavior in the 6 hydroxydopamine (6-OHDA) lesioned rat is the most utilized drug-induced paradigm for assessing functional efficacy in a rat model of Parkinson's disease (PD). Any clinically predictive animal model of PD should include a positive response to l dopa, the standard pharmacotherapy for PD. However, the acute interaction between L-dopa and apomorphine has never been studied to determine if L-dopa pretreatment could reduce apomorphine-induced rotational behavior in a 6-OHDA lesioned rat. The present study was designed to explore whether, indeed, pretreatment with subrotational doses of L-dopa could inhibit apomorphine-induced rotations. The data indicate that L-dopa significantly reduced apomorphine-induced rotational behavior only at one dose (5.0 mg/kg) for 12 min. Based on these and other data, it is concluded that although the apomorphine-induced rotational paradigm may continue to be utilized as one additional indicator of efficacy in the 6-OHDA rat model of PD, it is not in itself a completely valid functional assay. PMID- 10683288 TI - Chronic, selective forebrain responses to excitotoxic dorsal horn injury. AB - Intraspinal injection of the AMPA/metabotropic receptor agonist quisqualic acid (QUIS) results in excitotoxic injury which develops pathological characteristics similar to those associated with ischemic and traumatic spinal cord injury (SCI) (R. P. Yezierski et al., 1998, Pain 75: 141-155; R. P. Yezierski et al., 1993, J. Neurotrauma 10: 445-456). Since spinal injury can lead to partial or complete deafferentation of ascending supraspinal structures, it is likely that secondary to the disruption of spinal pathways these regions could undergo significant reorganization. Recently, T. J. Morrow et al. (Pain 75: 355-365) showed that autoradiographic estimates of regional cerebral blood flow (rCBF) can be used to simultaneously identify alterations in the activation of multiple forebrain structures responsive to noxious formalin stimulation. Accordingly, we examined whether excitotoxic SCI produced alterations in the activation of supraspinal structures using rCBF as a marker of neuronal activity. Twenty-four to 41 days after unilateral injection of QUIS into the T12 to L3 spinal segments, we found significant increases in the activation of 7 of 22 supraspinal structures examined. As compared to controls, unstimulated SCI rats exhibited a significant bilateral increase in rCBF within the arcuate nucleus (ARC), the hindlimb region of S1 cortex (HL), parietal cortex (PAR), and the thalamic posterior (PO), ventral lateral (VL), ventral posterior lateral (VPL), and ventral posterior medial (VPM) nuclei. All structures showing significantly altered rCBF are associated with the processing of somatosensory information. These changes constitute remote responses to injury and suggest that widespread functional changes occur within cortical and subcortical regions following injury to the spinal cord. PMID- 10683289 TI - Is Fos protein expressed by dying striatal neurons after immature hypoxic ischemic brain injury? AB - The transient induction of mRNA for the immediate-early gene c-fos has been reported following hypoxic-ischemic brain injury in the immature brain. However, no studies have examined the temporal expression of Fos protein, which is the functionally relevant product of c-fos gene expression. Increased expression of Fos protein has been linked to cell death. We therefore examined whether Fos protein is expressed by dying neurons after immature hypoxic-ischemic brain injury. A well characterized immature rat model of hypoxic-ischemic injury at postnatal day (PN) 7 was used. Three hypoxic-ischemic and three normoxic control pups were studied per time point (i.e., 0, 2, 12, 24, 48, and 72 h posttreatment). Expression of Fos within striatal and other neurons was detected immunocytochemically. Fos protein was expressed within dying striatal neurons at 0-12 h after hypoxia-ischemia. However, detection was only seen in 2 of 17 hypoxic-ischemic pups. These 2 pups had >/=80% of their striatal neurons dying within their right, hypoxic-ischemic-exposed hemisphere. Fos protein expression after severe injury may, therefore, be a response to extraordinary or extreme stress. The absence of Fos protein expression in the majority of hypoxic-ischemic pups, which all exhibited striatal neuronal death, suggests that Fos expression is not necessary for cell death to occur. Therapies directed against Fos protein expression may therefore have limited usefulness in immature hypoxic-ischemic brain injury. PMID- 10683290 TI - Evidence that Cereport's ability to increase permeability of rat gliomas is dependent upon extent of tumor growth: implications for treating newly emerging tumor colonies. AB - Cereport (RMP-7) enhances delivery of chemotherapeutics into brain tumors by increasing the permeability of the glioma vasculature (i.e. , the blood-brain tumor barrier; BBTB). Its effect on brain tumors has consistently been more robust than that on normal brain. The present experiments tested the hypothesis that the ability of Cereport to increase the permeability of infiltrating glioma colonies increases as the glioma colonies develop, in situ. In an initial preliminary experiment, the significant and selective effects of Cereport in tumor tissue and brain surrounding tumor were verified using [(14)C]carboplatin as a marker, 8 days after implantation of 50,000 RG2 cells. A second preliminary experiment established that the number of tumor cells initially seeded influences the growth rate of the tumor mass. Tumors seeded with 50,000 cells were larger than those seeded with 25,000 cells 3, 5, and 8 days after implantation. Next, the hypothesis that the extent of tumor growth increases Cereport's effects on the BBTB was tested by measuring the concentration of radiolabeled carboplatin in the tumor when 50,000 cells were implanted 3, 8, or 13 days prior to the experiment. While a reliable, approximately twofold increase in carboplatin concentration was seen in the 8- and 13-day-old tumors, no significant effect of Cereport was observed in the tumors that developed only 3 days, in situ. Finally, another test of the hypothesis was made by comparing Cereport's effects on 8-day old tumors initially seeded with either 50,000 or 25,000 cells (the latter producing a smaller, more slowly developing, tumor mass). Again, significantly higher carboplatin concentrations were seen with Cereport in the 50,000 cell tumors (greater than two-fold increase), compared to the smaller, more slowly developing, 25,000 cell tumors (<30% increase). The tumor and its vasculature were characterized in additional rats implanted with RG2 cells using CD-31, laminin, and bradykinin B(2) receptor immunocytochemistry. Intense B(2) receptor staining was observed on cells within the parenchyma of normal brain and tumor but not on the vasculature of tumor or brain. An extensive network of CD-31 and laminin staining was seen within and around the tumors in all groups, indicating relatively rapid and robust changes in vascularity in response to the gliomas. However, no consistent difference in vascularity between groups was observed to account for the uptake differences seen with Cereport. Collectively, these data offer initial preclinical empirical support for the hypothesis that Cereport's effects on tumor permeability increase as the tumor grows, which we further hypothesize is likely related to features of vascular development within the tumor independent of numbers or general morphology of vessels. If a similar phenomenon is shown to occur with infiltrating colonies from spontaneously forming gliomas in humans or from newly emerging metastases in brain, these data could impact the design and conduct of future trials using approaches intended to enhance delivery of chemotherapeutics through increased permeability of the tumor vascular barrier. PMID- 10683291 TI - P75 neurotrophin receptor in the nucleus basalis of meynert in relation to age, sex, and Alzheimer's disease. AB - In a previous study we showed that the staining of tyrosine kinase receptors (trks), which are high-affinity neurotrophin receptors (NTRs), is strongly diminished in the nucleus basalis of Meynert (NBM) of Alzheimer's disease (AD) patients, which may explain the lack of effect of NGF therapy in AD patients so far. Since the literature regarding the expression of low-affinity NTRs was rather controversial, the aim of the present study was to examine (i) possible changes in the staining of low-affinity NTRs, i.e., p75 in the human NBM, an area that is severely affected in AD; and (ii) alterations of these receptors in relation to risk factors for AD, e. g., age, sex, and menopause. Brain material of 31 controls and 30 AD patients was obtained at autopsy, embedded in paraffin, and stained immunocytochemically. Using an image analysis system, we quantified p75 immunoreactivity in both cell bodies and fibers at the level of the NBM. Our results showed a significant diminishment of p75 immunoreactivity in both cell bodies and fibers of NBM neurons in AD. We did not find any relationship between age or sex and the expression of p75 receptor in cell bodies. However, there was a clearly positive relationship between age and fiber staining in AD patients which suggests the occurrence of a p75 transport disorder as an early event in the process of AD. These observations and the earlier reported decreased staining of trk receptors show that degeneration of NBM neurons in AD is associated with a decreased neurotrophin responsiveness of NBM neurons in AD and that therapeutic strategies should be directed toward upregulation of receptors or facilitation of transport before an effect of neurotrophins in AD may be expected. PMID- 10683292 TI - Intracerebral implantation of NGF-releasing biodegradable microspheres protects striatum against excitotoxic damage. AB - Intrastriatal implantation of genetically modified cells synthesizing nerve growth factor (NGF) constitutes one way to obtain a long-term supply of this neurotrophic factor and a neuronal protection against an excitotoxic lesion. We have investigated if NGF-loaded poly(d,l-lactide-co-glycolide) microspheres could represent an alternative to cell transplantations. These microspheres can be implanted stereotaxically and locally release the protein in a controlled and sustained way. In order to test this paradigm, the NGF release kinetics were characterized in vitro using radiolabeled NGF, immunoenzymatic assay, and PC-12 cells bioassay and then in vivo after implantation in the intact rat striatum. These microspheres were thus implanted into the rat striatum 7 days prior to infusing quinolinic acid. Control animals were either not treated or implanted with blank microspheres. The extent of the lesion and the survival of ChAT-, NADPH-d-, and DARPP-32-containing neurons were analyzed. In vitro studies showed that microspheres allowed a sustained release of bioactive NGF for at least 1 month. Microspheres implanted in the intact striatum still contained NGF after 2.5 months and they were totally degraded after 3 months. After quinolinic acid infusion, the lesion size in the group treated with NGF-releasing microspheres was reduced by 40% when compared with the control groups. A marked neuronal sparing was noted, principally concerning the cholinergic interneurons, but also neuropeptide Y/somatostatin interneurons and GABAergic striatofuge neurons. These results indicate that implantation of biodegradable NGF-releasing microspheres can be used to protect neurons from a local excitotoxic lesion and that this strategy may ultimately prove to be relevant for the treatment of various neurological diseases. PMID- 10683293 TI - Changes in urinary bladder neurotrophic factor mRNA and NGF protein following urinary bladder dysfunction. AB - Spinal cord injury and cyclophosphamide-induced cystitis dramatically alter lower urinary tract function and produce neurochemical, electrophysiological, and anatomical changes that may contribute to reorganization of the micturition reflex. Mechanisms underlying this neural plasticity may involve alterations in neurotrophic factors in the urinary bladder. These studies have determined neurotrophic factors in the urinary bladder that may contribute to reorganization of the micturition reflex following cystitis or spinal cord injury. A ribonuclease protection assay was used to measure changes in urinary bladder neurotrophic factor mRNA (betaNGF, BDNF, GDNF, CNTF, NT-3, and NT-4) following spinal cord injury (acute/chronic) or cyclophosphamide-induced cystitis (acute/chronic). The correlation between urinary bladder nerve growth factor mRNA and nerve growth factor protein expression was also determined. Each experimental paradigm resulted in significant (P 1000 pg/ml IL-8 and >400 pg/ml MCP-1 per million cells), while in the cell lines EOL-1, KASUMI-1 and KG-1 only 10-100 pg/ml IL-8 and MCP-1 were detected. Tetradecanoyl phorbol acetate (TPA) strongly increased the IL-8 and MCP-1 amounts in the culture supernatants of all five cell lines. The TPA-induced NB-4 produced the largest amounts of both chemokines (>40,000 pg/ml). The strongest induction was seen in EOL-1 (>100-fold increase). Besides TPA, tumor necrosis factor-alpha (TNF alpha) also distinctively enhanced IL-8 and MCP-1 production. The calcium ionophore A 23187 and thapsigargin, an inhibitor of the Ca(2+)-ATPase, differentially induced IL-8 and MCP-1 secretion in the cell lines investigated, suggesting that, at least in some cell lines, intracellular free Ca(2+) might be important for chemokine secretion. Dexamethasone significantly prevented the IL-8 and MCP-1 production of stimulated cells, emphasizing the potent anti-inflammatory property of glucocorticoids. Similarly, the protein kinase inhibitor staurosporine clearly decreased the TPA-induced chemokine secretion in NB-4 cells, indicating the involvement of protein kinases in the signal transduction pathway which leads to enhanced chemokine secretion. PMID- 10683320 TI - Proteolytic processing of Marburg virus glycoprotein. AB - Processing of the transmembrane glycoprotein (GP) of Marburg virus involved the conversion of an endo H-sensitive, ER-specific form into an endo H-resistant, Golgi-specific precursor that was cleaved into GP(1) and GP(2). Cleavage was mediated by furin or another subtilisin-like endoprotease with similar substrate specificity as indicated by mutational analysis of the cleavage site and inhibition using peptidyl chloromethylketones. Mature GP consisted of disulfide linked GP(1) and GP(2) subunits. PMID- 10683321 TI - Canine cyclin T1 rescues equine infectious anemia virus tat trans-activation in human cells. AB - Human immunodeficiency virus-1 Tat protein and human Cyclin T1 mediate transcriptional activation by enhancing the elongation efficiency of RNA polymerase II. Activation of transcription of the related equine infectious anemia virus (EIAV) requires a similar protein known as eTat, which does not function in human cells. Expression of equine Cyclin T1 in human cells rescues eTat function, suggesting a general mechanism of transcription activation among lentiviruses. Here we present the cloning of Cyclin T1 from canine D17 osteosarcoma cells, which support EIAV transactivation, and show that canine Cyclin T1 confers eTat transactivation to human cells. A two-amino-acid change, from 79-proline-glycine-80 to 79-histidine-arginine-80, confers on the human Cyclin T1 the ability to cooperate with eTat in transcriptional activation. These findings suggested that the regions of Cyclin T1 that interact with lentiviral Tat proteins and TAR RNA elements form an extended domain, which very likely has a conserved fold. PMID- 10683322 TI - Peripheral blood mononuclear cells of HIV- and HCV-antibody-positive individuals contain HCV RNA but No HCV DNA despite evidence for reverse transcription of HIV RNA into DNA. AB - Following reports of the finding of cDNA of RNA viruses in cells containing an endogenous retrovirus-encoded reverse transcriptase, we looked for the presence of hepatitis C virus (HCV) DNA in peripheral blood mononuclear cells (PBMC) of injecting drug users seropositive for both HCV and human immunodefiency virus (HIV). We tested serial PBMC samples from four HCV infected individuals; one was seronegative for HIV, two seroconverted for HIV during follow-up, and one was seropositive for HIV throughout the study period. HCV RNA was found in PBMC and plasma samples at all time points tested. Similarly, HIV RNA was found in all PBMC and plasma samples following HIV seroconversion. In contrast, no HCV DNA was detected in any PBMC sample, whereas HIV DNA was found in all tested PBMC samples following HIV seroconversion, indicative of active HIV reverse transcriptase in these PBMC samples. These results do not support the hypothesis that HCV viraemia is related to retrotranscription of the HCV RNA genome into DNA in peripheral blood mononuclear cells coinfected with HIV. The potential of HIV RT to retrotranscribe HCV RNA into DNA awaits studies of liver cells coinfected with HCV and HIV. PMID- 10683323 TI - Establishment of latent herpes simplex virus type 1 infection in resistant, sensitive, and immunodeficient mouse strains. AB - Productive infection with herpes simplex virus (HSV) type 1 is limited by both innate and adaptive immune mechanisms. The purpose of the current study was to determine whether these mechanisms also play a role in the establishment of latent HSV infection. First we examined the trigeminal ganglia (TG) of severe combined immunodeficiency (SCID), interferon-gamma knockout (GKO), and beige (a strain deficient in natural killer cell activity) mice following ocular inoculation with HSV. Although infection of SCID mice was invariably lethal, we consistently found latently infected neurons in the TG of these animals at 2-4 days postinoculation. HSV infection of GKO and beige mice, while not lethal, was characterized by a greater number of productively infected TG neurons and/or a delay in the time to peak productive infection compared to C57BL/6 controls. However, as assayed by both in situ hybridization for LAT expression and quantitative PCR (Q-PCR) for viral DNA, we found that HSV established a latent infection in GKO and beige mice as efficiently as in C57BL/6 controls. We subsequently examined the TG of "HSV-sensitive" strains of mice (Swiss-Webster, CBA, and BALB/c) following ocular infection with HSV. At the peak of acute ganglionic infection the number of productively infected TG neurons in each of these mouse strains was about sevenfold greater than in the "HSV-resistant" strain C57BL/6, consistent with previously reported differences in susceptibility to lethal challenge with HSV. However, as assayed by both in situ hybridization for LAT and Q-PCR for viral DNA, we found that HSV established a latent infection in Swiss-Webster, CBA, and BALB/c mice as efficiently as in C57BL/6 controls. We conclude that HSV efficiently establishes latent infection in the TG of mice in the absence of innate and adaptive immune mechanisms that are essential for limiting productive viral infection. PMID- 10683324 TI - Genetic dissection of the multiple functions of alfalfa mosaic virus coat protein in viral RNA replication, encapsidation, and movement. AB - Coat protein (CP) of alfalfa mosaic virus (AMV) binds as a dimer to the 3' termini of the three genomic RNAs and is required for initiation of infection, asymmetric plus-strand RNA accumulation, virion formation, and spread of the virus in plants. A mutational analysis of the multiple functions of AMV CP was made. Mutations that interfered with CP dimer formation in the two-hybrid system had little effect on the initiation of infection or plus-strand RNA accumulation but interfered with virion formation and reduced or abolished cell-to-cell movement of the virus in plants. Six of the 7 basic amino acids in the N-terminal arm of CP (positions 5, 6, 10, 13, 16, and 25) could be deleted or mutated into alanine without affecting any step of the replication cycle except systemic movement in plants. Mutation of Arg-17 interfered with initiation of infection (as previously shown by others) and cell-to-cell movement of the virus but not with plus-strand RNA accumulation or virion formation. The results indicate that in addition to the RNA-binding domain, different domains of AMV CP are involved in initiation of infection, plus-strand RNA accumulation, virion formation, cell to-cell movement, and systemic spread of the virus. PMID- 10683325 TI - The HTLV receptor is a widely expressed protein. AB - The receptor for human T-cell leukemia virus type 1 (HTLV-1) was found to be expressed on a broad range of cell lines derived from multiple species. Receptor expression was assessed using human immunodeficiency virus type 1 particles, pseudotyped with the HTLV-1 envelope glycoprotein, and expressing luciferase under the control of an SV40 enhancer and promoter. Infection by pseudotyped virus was blocked with neutralizing antibodies to HTLV-1, and infection was dependent on the presence of the cleavage and fusogenic sequences in the envelope protein precursor. Trypsin treatment of susceptible target lymphocytes reduced entry. Entry was partially resistant to ammonium chloride. PMID- 10683326 TI - Japanese encephalitis DNA vaccine candidates expressing premembrane and envelope genes induce virus-specific memory B cells and long-lasting antibodies in swine. AB - Swine are an important amplifier of Japanese encephalitis (JE) virus in the paradomestic environment. In this study, two JE DNA vaccine candidates were evaluated for immunogenicity in swine. Both vaccine plasmids encode a cassette consisting of the signal of premembrane (prM), prM, and envelope (E) coding regions of JE virus. One plasmid, designated pcJEME, is based on a commercial vector (pcDNA3), whereas the other plasmid, designated pNJEME, is based on a vector (pNGVL4a) designed to address some of the safety concerns of DNA vaccine use. No differences were detected in the immunogenicity of these two plasmids in mice or swine. Swine immunized with the DNA vaccines at a dose of 100 to 450 microgram at an interval of 3 weeks developed neutralizing and hemagglutination inhibitory (HAI) antibody titers of 1:40 to 1:160 at 1 week after the second immunization. However, swine administered two doses of a commercial JE vaccine (formalin-inactivated virus preparation; JEVAX-A) developed low (1:10) or undetectable antibody responses after their boost. Interestingly, serum antibody titers elicited by DNA vaccines in swine were higher than those detected in mice. Eight days after boosting with viral antigen (JEVAX-A) to detect an anamnestic response, swine immunized two times with the DNA vaccine showed a >100-fold elevation in HAI titer, indicating a strong recall of antibody response. Swine maintained detectable levels of HAI antibody for at least 245 days after two immunizations with a DNA vaccine. These results indicate that these DNA vaccines are able to induce virus-specific memory B cells and long-lasting antibodies in swine, which were of higher levels than those obtained with a commercial formalin inactivated JE vaccine. PMID- 10683327 TI - Characterization of an overexpressed spindle protein during a baculovirus infection. AB - The nucleopolyhedrovirus CfDEFNPV contains a gene encoding a viral protein, which accumulates as bipyramidal inclusion bodies (spindles) in the cytoplasm of infected cells. The spindles appear as early as 24 h postinfection, approximately 1 day earlier than viral occlusion bodies (OBs). Purification and characterization of the spindle protein was complicated by the fact that the OBs copurified with the spindles. We therefore modified CfDEFNPV by replacing the polyhedrin gene (plh) with a cassette containing the green fluorescent protein (GFP) gene. The recombinant virus did not produce OBs; however, the synthesis and morphogenesis of the spindles were not altered. When analyzed by SDS-PAGE, the spindles produced a 50-kDa protein, which was termed spindlin. Tunicamycin inhibition and endoglycosidase studies showed that spindlin was glycosylated. The N-terminus of spindlin was sequenced and its gene (gp50) was located on the viral genome. The gene was cloned and sequenced. Homologs of gp50 were found in several baculoviruses as well as in entomopoxviruses (EPV). In the latter virus, the homologous gene is that of fusolin, which also encodes a protein that forms spindle-shaped inclusion bodies in the cytoplasm of infected cells. Immunoblot analysis indicated that spindlin and fusolin were not serologically related, even though they share conserved polypeptide domains. Sequence analysis showed that gp50 of CfDEFNPV contains two late promoter motifs (TTAAG) in its 5' flanking region. Both were used, but the proximal motif (-14 to -18 nt relative to the ATG) was the primary sequence from which most of the mRNA was initiated. When gp50 was cloned in a heterologous baculovirus expression system, spindlin was synthesized, although the spindles were irregular in shape. This suggested that the spindle structure may be species-specific or it may require more than one gene product for its morphogenesis. PMID- 10683328 TI - Loss of ATM function enhances recombinant adeno-associated virus transduction and integration through pathways similar to UV irradiation. AB - Ataxia telangiectasia is caused by a genetic defect in the ATM gene that results in altered cellular sensitivity to DNA-damaging agents such as gamma-irradiation. ATM deficiency is associated with an increased incidence of neurological disorders, immune deficiency, and cancer. In this report we demonstrate that recombinant adeno-associated virus (rAAV) gene transfer in ATM-deficient fibroblasts is significantly enhanced over normal fibroblast cell lines. This enhancement of rAAV transduction in AT cells is correlated with an increased abundance of circular form rAAV genomes, as well as a higher number of integrated head-to-tail concatamer proviral genomes. Studies evaluating AAV trafficking using Cy3-labeled virus suggest that a nuclear mechanism is responsible for increased rAAV transduction in AT cells, because binding, endocytosis, and nuclear trafficking of virus are unaffected by the AT phenotype. Additionally, the profile of rAAV transduction after UV irradiation is significantly blunted in AT cells, suggesting that the level of DNA repair enzymes normally associated with UV augmentation of viral transduction may already be maximally elevated. These results further expand our understanding of genes involved in rAAV transduction. PMID- 10683329 TI - Construction of a selectable nef-defective live-attenuated human immunodeficiency virus expressing Escherichia coli gpt gene. AB - We have developed a replication-competent human immunodeficiency virus (HIV) carrying a selective marker that can be used in vivo. This recombinant virus (Z6 Delta nef gpt) was generated by replacing the 5' half of the HIV nef gene with the Escherichia coli guanine phosphoribosyl transferase gene (gpt). This new vector can express the gpt product on infection and works as a positive selective marker for mycophenolic acid (MPA) resistance, a potent immunosuppressive drug used in organ rejection therapy. Conversely, gpt expression also served as a negative selectable marker, since its intracellular expression induces host-cell susceptibility to 6-thioxantine (6-TX), a nucleotide analog that is toxic to the infected cell under these conditions. In this manner, we could suppress the recombinant virus replication through 6-TX selection in both transformed cells and primary human peripheral blood mononuclear cells (PBMCs), suggesting the vector's potential as a model for a new live-attenuated vaccine approach against HIV. PMID- 10683330 TI - Porcine endogenous retroviruses inhibit human immune cell function: risk for xenotransplantation? AB - Transgenic pigs are currently the most favored potential source of organs for xenotransplantation. Like all mammalian species they all harbor endogenous retroviruses in their genome. These porcine endogenous retroviruses (PERVs) are produced from several primary cells and cell lines and are able to infect human cells. Here we demonstrate that different pig strains and different animals of one strain differ in their ability to produce PERVs from normal blood cells. We report that purified PERV particles show a protein pattern typical for type C retroviruses and are antigenically related to mammalian leukemia viruses. Like most retroviruses, purified PERVs and peptides derived from the highly conserved immunosuppressive domain of their transmembrane envelope protein inhibit human immune cell functions. This indicates that high titer replication of PERVs in the transplant recipient could therefore lead to an immunodeficiency disease. PMID- 10683331 TI - The kinetics of specific immune responses in rhesus monkeys inoculated with live recombinant BCG expressing SIV Gag, Pol, Env, and Nef proteins. AB - Development of an effective preventive or therapeutic vaccine against HIV-1 is an important goal in the fight against AIDS. Effective virus clearance and inhibition of spread to target organs depends principally on the cellular immune response. Therefore, a vaccine against HIV-1 should elicit virus-specific cytotoxic lymphocyte (CTL) responses to eliminate the virus during the cell associated stages of its life cycle. The vaccine should also be capable of inducing immunity at the mucosal surfaces, the primary route of transmission. Recombinant Bacille Calmette-Guerin (BCG) expressing viral proteins offers an excellent candidate vaccine in view of its safety and ability to persist intracellularly, resulting in the induction of long-lasting immunity and stimulation of the cellular immune response. BCG can be administered orally to induce HIV-specific immunity at the mucosal surfaces. The immunogenicity of four recombinant BCG constructs expressing simian immunodeficiency virus (SIV) Gag, Pol, Env, and Nef proteins was tested in rhesus macaques. A single simultaneous inoculation of all four recombinants elicited SIV-specific IgA and IgG antibody, and cellular immune responses, including CTL and helper T cell proliferation. Our results demonstrate that BCG recombinant vectors can induce concomitant humoral and cellular immune responses to the major proteins of SIV. PMID- 10683332 TI - Deletion mapping of the potyviral helper component-proteinase reveals two regions involved in RNA binding. AB - The Potyvirus helper component-proteinase (HC-Pro) binds nonspecifically to single-stranded nucleic acids with a preference for RNA. To delineate the regions of the protein responsible for RNA binding, deletions were introduced into the full-length Potato potyvirus Y HC-Pro gene carried by an Escherichia coli expression vector. The corresponding proteins were expressed as fusions with the maltose-binding protein, purified, and assayed for their RNA-binding capacity. The results obtained by UV cross-linking and Northwestern blot assays demonstrated that the N- and C-terminal regions of HC-Pro are dispensable for RNA binding. They also revealed the presence of two independent RNA-binding domains (designated A and B) located in the central part of HC-Pro. Domain B appears to contain a ribonucleoprotein (RNP) motif typical of a large family of RNA-binding proteins involved in several cellular processes. The possibility that domain B consists of an RNP domain is discussed and suggests that HC-Pro could constitute the first example of a plant viral protein belonging to the RNP-containing family of proteins. PMID- 10683333 TI - Expression of human immunodeficiency virus type 1 Gag protein precursor and envelope proteins from a vesicular stomatitis virus recombinant: high-level production of virus-like particles containing HIV envelope. AB - Recombinant vesicular stomatitis viruses have been developed as high-level expression vectors which serve as effective vaccine vectors in animals (Roberts et al., 1998, J. Virol. 72, 4704-4711; Roberts et al., 1999, J. Virol. 73, 3723 3732). Here we show that two genes can be expressed simultaneously from a single, live-attenuated VSV recombinant. The genes used encode the Pr55(gag) protein precursor of HIV-1 (1.7-kb gene) and an HIV-1 envelope (Env) protein (2.4 kb gene). Our results show that VSV can accommodate up to a 40% increase in genome size with only a threefold reduction in virus titer. Recombinants expressing the Pr55(gag) protein precursor with or without Env protein produced abundant HIV virus-like particles (VLPs) in addition to bullet-shaped VSV particles. HIV Env protein expressed from a VSV recombinant also expressing Gag was specifically incorporated into the HIV VLPs but not into the VSV particles. In contrast, VSV G protein was found in both VSV particles and in HIV VLPs. Such VSV/HIV recombinants producing HIV VLPs with Env protein could be an effective source of HIV-like particles inducing both cellular and antibody-mediated immunity to HIV 1. PMID- 10683334 TI - Limitations of in vivo IL-12 supplementation strategies to induce Th1 early life responses to model viral and bacterial vaccine antigens. AB - The limited induction of Th1 and cytotoxic immune responses is regarded as the main reason for the increased susceptibility to intracellular microorganisms in early life. Recently, in vitro IL-12 supplementation was shown to enhance the limited IFN-gamma release of measles-specific infant T cells. Using a series of IL-12 delivery systems, we show here that in vivo IL-12 supplementation may enhance early life murine Th1 responses to two model vaccine antigens, measles virus hemagglutinin and tetanus toxin peptide. However, this required multiple repeat injections of recombinant rIL-12, which were poorly tolerated in young mice. Local IL-12 delivery by an IL-12 expressing canarypox vector proved safe but failed to modulate vaccine responses. An IL-12 DNA plasmid or a CD40L DNA plasmid efficiently enhanced neonatal Th1 responses to measles hemagglutinin DNA vaccine. However, both plasmids only enhanced Th1 responses to DNA and not to peptide, protein, or live viral vaccines. Thus, inducing adult-like Th1 responses may be achieved in vivo by inducing (CD40L) or substituting for (IL-12 supplementation) optimal activation of neonatal APC. However, these immunomodulatory effects appear limited to certain antigen-presentation approaches and may not be broadly applicable to vaccines. PMID- 10683335 TI - Internal cleavage of hepatitis C virus NS3 protein is dependent on the activity of NS34A protease. AB - The nonstructural protein NS3 of the hepatitis C virus (HCV) is indispensable for virus replication and a multifunctional enzyme that contains three catalytic activities such as serine protease, helicase, and NTPase. Here, we demonstrated that the internal cleavage of the HCV NS3 protein occurs in various mammalian cells such as HepG2, COS-7, and NIH3T3. As is observed for the internal cleavage mechanism of the NS3 protein of dengue virus 2, the internal processing of HCV NS3 protein was catalyzed by the active NS3 serine protease and NS4A, but not NS3 alone. From the data acquired from extensive site-directed mutagenesis, we observed that the NS3 protein was internally cleaved at two different sites, FCH(1395) ||S(1396)KK and IPT(1428) ||S(1429)GD, within RNA helicase domain. The internal cleavage of NS3 protein by NS34A protease was also confirmed in a different isolate of HCV-1b strain. In addition, in vitro transforming assays demonstrated that the internal cleavage product of NS3, NS3a-1, appeared to have higher oncogenic potential than does intact NS3. Taken together, our results suggest that the internal cleavage of NS3 may be associated with the replication and oncogenesis of HCV. PMID- 10683336 TI - Adenovirus endopeptidase hydrolyses human squamous cell carcinoma antigens in vitro but not ex vivo. AB - The serpins SCCA1 and SCCA2 are highly expressed in the epithelium of the conducting airways, a common site of infection by group C adenoviruses, such as human adenovirus type 2 (Ad2). Based on the common location we examined a possible interaction between them. In vitro experiments with recombinant proteins showed that SCCA1 inhibited the viral protease in a dose-dependent manner. Both serpins were cleaved in a manner consistent with hydrolysis within their reactive site loop, without the formation of an SDS-resistant complex, as in the case of papain. Infection of SCCA1-expressing cells did not result in the cleavage of SCCA1, nor was the yield of infectious virus affected as compared to SCCA1 negative parental cells. This may be due to differential localization, the serpin being cytoplasmic and viral protease being nuclear. Surprisingly, however, virus infection, which tends to inhibit host protein synthesis, caused a significant increase in SCCA1 expression well into the late phase of infection. PMID- 10683337 TI - Cellular expression of alphaherpesvirus gD interferes with entry of homologous and heterologous alphaherpesviruses by blocking access to a shared gD receptor. AB - Several human and animal alphaherpesviruses can enter cells via human herpesvirus entry mediator C (HveC), a receptor for viral glycoprotein D (gD). In previous studies with cells expressing unknown entry mediators, cellular expression of alphaherpesvirus gD was shown to inhibit entry of the homologous virus and sometimes also of heterologous alphaherpesviruses. To investigate the mechanism of gD-mediated interference and the basis for cross-interference among alphaherpesviruses, HveC was expressed in cells as the sole entry mediator, in the presence or absence of one of the gDs encoded by herpes simplex virus type 1, pseudorabies virus, or bovine herpesvirus type 1. Cells expressing HveC alone were highly susceptible to entry of all three viruses, whereas cells coexpressing HveC and any one of the gDs were at least partially resistant to infection by each virus. Coexpression of gD with HveC did not cause reduced levels of cell surface HveC but the HveC had reduced ability to bind to exogenous gD. Coimmunoprecipitation experiments revealed that HveC was complexed with gD in lysates of cells expressing both. Thus, cellular expression of gD can interfere with alphaherpesvirus entry by blocking ligand-binding sites of the gD receptor(s) used for entry and cross-interference can occur because different forms of alphaherpesvirus gD can compete for shared entry receptors. PMID- 10683338 TI - A role for bovine herpesvirus 1 (BHV-1) glycoprotein E (gE) tyrosine phosphorylation in replication of BHV-1 wild-type virus but not BHV-1 gE deletion mutant virus. AB - Bovine herpesvirus 1 (BHV-1), an alphaherpesvirus, is a major pathogen that causes respiratory and reproductive infections. We observed tyrosine phosphorylation of a 95-kDa viral protein and dephosphorylation of 55- and 103 kDa cellular proteins during the course of BHV-1 infection. We demonstrated BHV-1 glycoprotein E (gE) to be the tyrosine phosphorylated viral protein by immunoprecipitation. Inhibition of phosphorylation of BHV-1 gE by tyrosine kinase inhibitors genistein and tyrphostin AG1478 substantially lowered the viral titer in Madin-Darby bovine kidney cells. The decrease in viral titer was directly proportional to the decrease in phosphorylation of the BHV-1 gE. Interestingly, these kinase inhibitors did not inhibit the replication of the BHV-1 gE deletion mutant virion (BHV-1gEDelta3.1). Our findings suggest that the wild-type BHV-1, with a functional gE protein, uses a different pathway of signaling events than the BHV-1 gE deletion mutant in replication. Our results indicate that the tyrosine phosphorylation of the cytoplasmic tail of BHV-1 gE is an important post translational modification of the functional protein. An application of this study may be the use of tyrosine kinase inhibitors in controlling the BHV-1 infection. PMID- 10683339 TI - Distinct transcriptional and functional properties of the R transactivator gene orf50 of the transforming herpesvirus saimiri strain C488. AB - The transformation-associated region of herpesvirus saimiri strains is variable, whereas other parts of the virus genome are highly conserved. However, we observed considerable interstrain sequence divergence of the early viral regulatory orf50 gene, which encodes the R transactivator, a homolog of Epstein Barr virus BRLF1. The orf50 gene of strain C488 was transcribed at low abundance during lytic infection, whereas antisense transcripts were simultaneously expressed at high levels. A spliced variant, orf50a, was detectable by RT-PCR and RNase protection assays in stimulated C488-transformed, nonpermissive human T cells. In contrast to strain A11, the short, unspliced orf50b form of C488 displayed complete transactivation capability on the orf6 and orf57 promoters. In summary, there are unexpected structural and functional differences between the orf50 genes of herpesvirus saimiri strains, which differ in their capability to transform human T lymphocytes. PMID- 10683340 TI - The Epstein-Barr virus latent membrane protein 2A PY motif recruits WW domain containing ubiquitin-protein ligases. AB - Latent membrane protein 2A (LMP2A) is expressed in latent Epstein-Barr virus (EBV) infection. LMP2A functions to downregulate B-cell signal transduction and viral reactivation from latency in EBV-immortalized B cells in vitro, and acts to provide B cells with both a survival and developmental signal in vivo. Identification of proteins associated with LMP2A is important for elucidation of the mechanism that LMP2A employs to regulate B-cell signal transduction and EBV latency. LMP2A is constitutively tyrosine phosphorylated and is associated with protein tyrosine kinases such as Lyn and Syk when specific LMP2A tyrosines are phosphorylated. The amino-terminal domain of LMP2A includes multiple proline-rich regions, which may provide binding sites for proteins containing SH3 or WW domains. In this study, we demonstrate that four cellular proteins bind specifically to two PPPPY (PY) motifs present within the LMP2A amino-terminal domain. Protein microsequence analysis determined that three of these proteins were AIP4, WWP2/AIP2, and Nedd4. All of these proteins are members of the Nedd4 like ubiquitin-protein ligases family and have conserved domains including the C2, WW, and ubiquitin-protein ligase domain. The mutation of both PY motifs completely abolished binding activity of these proteins to LMP2A and the interaction of AIP4 and WWP2 with LMP2A was confirmed in cell lines expressing LMP2A, WWP2, and AIP4. Furthermore, a reduction in the level of Lyn and the rapid turnover of LMP2A and Lyn were observed in LMP2A-expressing cells. These findings suggest that LMP2A recruits Nedd4-like ubiquitin-protein ligases and B-cell signal transduction molecules, resulting in the degradation of LMP2A and Lyn by a ubiquitin-dependent mechanism. This provides a new means by which LMP2A may modulate B-cell signal transduction. PMID- 10683341 TI - Genetic analysis of the cell-to-cell movement of beet yellows closterovirus. AB - A beet yellows closterovirus (BYV) variant expressing green fluorescent protein and leaves of BYV local lesion host Claytonia perfoliata were used to reveal genetic requirements for BYV cell-to-cell movement in leaf epidermis and mesophyll. A series of mutations targeting genes that are not involved in amplification of the viral positive-strand RNA was analyzed. The products of genes coding for a 6-kDa hydrophobic protein (p6) and a 64-kDa protein (p64), as well as for minor and major capsid proteins, were found to be essential for intercellular translocation of BYV. In a previous work, we have demonstrated that the BYV HSP70-homolog (HSP70h) also plays a critical role in viral movement (V. V. Peremyslov, Y. Hagiwara, and V. V. Dolja, 1999, Proc. Natl. Acad. Sci. USA, 96, 14771-14776). Altogether, a unique protein quintet including three dedicated movement proteins (p6, p64, and HSP70h) and two structural proteins is required to potentiate the cell-to-cell movement of a closterovirus. The corresponding BYV genes are clustered in a block that is conserved among diverse representatives of the family Closteroviridae. PMID- 10683342 TI - Dihydrofolate reductase from Kaposi's sarcoma-associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the first human virus known to encode dihydrofolate reductase (DHFR), an enzyme required for nucleotide and methionine biosynthesis. We have studied the purified KSHV-DHFR enzyme in vitro and analyzed its expression in cultured B-cell lines derived from primary effusion lymphoma (PEL), an AIDS-associated malignancy. The amino acid sequence of KSHV-DHFR is most similar to human DHFR (hDHFR), but the viral enzyme contains an additional 23 amino acids at the carboxyl-terminus. The viral DHFR, overexpressed and purified from E. coli, was catalytically active in vitro. The K(m) of KSHV-DHFR for dihydrofolate (FH(2)) was 2.4 microM, which is significantly higher than the K(m) of recombinant hDHFR (rhDHFR) for FH(2) (390 nM). K(m) values for NADPH were similar for the two enzymes, about 1 microM. KSHV DHFR was inhibited by folate antagonists such as methotrexate (K(i): 200 pM), aminopterin (K(i): 610 pM), pyrimethamine (K(i): 29 nM), trimethoprim (K(i): 2.3 microM), and piritrexim (K(i): 3.9 nM). In all cases, K(i) values for these folate antagonists were higher for KSHV-DHFR than for rhDHFR. The viral enzyme was expressed at levels two- to tenfold higher than hDHFR in PEL cell lines as an early lytic cycle gene. KSHV-DHFR mRNA and protein appeared from 6 to 24 h after chemical induction of the KSHV lytic cycle. Epitope-tagged KSHV-DHFR and rhDHFR both localized to the nucleus of transfected cells, while other KSHV nucleotide metabolism genes localized to the cytoplasm. DHFR activity was not essential for viral replication in cultured PEL cells. Since hDHFR was not detectable in peripheral blood mononuclear cells (PBMCs), KSHV-DHFR may function to provide increased DHFR activity in vivo in infected cells that have little or none of their own enzyme. PMID- 10683343 TI - A chloroplastic RNA polymerase resistant to tagetitoxin is involved in replication of avocado sunblotch viroid. AB - Avocado sunblotch viroid (ASBVd), the type species of the family Avsunviroidae, replicates and accumulates in the chloroplast. Two main chloroplastic RNA polymerases have been described: the plastid-encoded polymerase (PEP) with a multisubunit structure similar to the Escherichia coli enzyme and a single-unit nuclear-encoded polymerase (NEP) resembling phage RNA polymerases. On a different basis, sensitivity to tagetitoxin, two major RNA polymerase activities, tagetitoxin sensitive (TS) and resistant (TR), have been found in plastids. The most plausible candidates for the TS and TR RNA polymerases are PEP and NEP, respectively. To gain an insight into the enzymology of the polymerization of ASBVd strands, purified chloroplast preparations from ASBVd-infected leaves were assayed for their in vitro ability to transcribe ASBVd RNAs together with some representative genes (psbA, 16SrDNA, accD, and rpoB) of the three classes of chloroplastic genes according to their promoter structure. High concentrations of alpha-amanitin had no effect on gene or on viroid transcription, but tagetitoxin (5-10 microM) prevented transcription of all these genes without affecting synthesis of ASBVd strands; only at higher tagetitoxin concentrations (50-100 microM) was a 25% inhibition observed. These results suggest that NEP is the RNA polymerase required in ASBVd replication, although the participation of another TR RNA polymerase from the chloroplast cannot be excluded. PMID- 10683344 TI - Circulation online only : february 22, 2000 PMID- 10683345 TI - Distinguishing mechanisms from markers of cardiac contractile dysfunction: more than 1 way to skin the cat of heart failure. PMID- 10683346 TI - Paraoxonase polymorphism (Gln192Arg) as a determinant of the response of human coronary arteries to serotonin. AB - Background-Oxidation of LDL plays a role in endothelial dysfunction. Paraoxonase, an enzyme present on HDL, protects LDL against oxidation. Paraoxonase activity is genetically determined in part, and 3 genotypes have been described with variable enzymatic activity. We hypothesized that the paraoxonase polymorphism might influence endothelial function. Methods and Results-Twenty-seven patients with clinical manifestations of coronary artery disease underwent provocative testing by intracoronary administration of serotonin. None of the coronary arteries studied had significant (>50%) stenosis. Ten patients had the QQ genotype and 17 had the QR genotype. At proximal segments, the mean percentage reduction in lumen diameter in response to serotonin was greater in QQ patients than in QR patients (10(-5) mol/L: P<0.05; 10(-4) mol/L: P<0.006). Similarly, at distal segments, constriction in response to serotonin was greater in QQ patients than in QR patients (10(-6) mol/L: P<0. 03; 10(-5) mol/L: P<0.07). Conclusions-These results suggest a higher synthesis or release of endothelium-derived relaxing factors to counteract the vasoconstrictor effect of serotonin in patients with the R allele. These findings provide evidence that the paraoxonase polymorphism may play a role in the regulation of coronary vasomotor tone. PMID- 10683347 TI - Hyperfibrinogenemia is associated with specific histocytological composition and complications of atherosclerotic carotid plaques in patients affected by transient ischemic attacks. AB - BACKGROUND: Epidemiological studies have demonstrated that hyperfibrinogenemia is an independent risk factor for cerebrovascular atherosclerosis. However, the underlying mechanisms are poorly understood. We studied whether hyperfibrinogenemia could modify the histological composition of atherosclerotic plaque and precipitate carotid thrombosis resulting from rupture of the plaque. METHODS AND RESULTS: We studied the histological composition of 71 carotid atherosclerotic plaques from patients who had undergone surgical endarterectomy after a first episode of transient ischemic attack. Patients were divided into 3 groups corresponding to the tertiles of plasma fibrinogen values. Hypercholesterolemia, hypertriglyceridemia, hypertension, diabetes, and smoking habit were also assessed. At the histological analysis, plaques of patients in the highest tertile of fibrinogen (>407 mg/dL) were characterized by a high incidence of thrombosis (66.7% of cases) compared with plaques of subjects in the lower (21.7%) (P=0.002) and middle (29. 2%) (P=0.009) tertiles. Plaque rupture was significantly associated with high fibrinogen levels (54.2%, P=0.003). Multivariate logistic regression indicated that hyperfibrinogenemia was an independent risk factor for a decrease in cap thickness (P=0.0005), macrophage foam cell infiltration of the cap (P=0.003), and thrombosis (P=0. 003). When the presence of other risk factors was accounted for, hyperfibrinogenemia remained an independent predictor of carotid thrombosis with an odds ratio of 5.83, compared with other risk factors. CONCLUSIONS: The results of the present study add to the evidence that hyperfibrinogenemia, independently of other risk factors, is associated with a specific histological composition of carotid atherosclerotic plaques that predisposes them to rupture and thrombosis. PMID- 10683348 TI - Early percutaneous coronary intervention, platelet inhibition with eptifibatide, and clinical outcomes in patients with acute coronary syndromes. PURSUIT Investigators. AB - BACKGROUND: Platelet glycoprotein (GP) IIb/IIIa antagonists prevent the composite end point of death or myocardial infarction (MI) in patients with acute coronary syndromes. There is uncertainty about whether this effect is confined to patients who have percutaneous coronary interventions (PCIs) and whether PCIs further prevent death or MI in patients already treated with GP IIb/IIIa antagonists. METHODS AND RESULTS: PURSUIT patients were treated with the GP IIb/IIIa antagonist eptifibatide or placebo; PCIs were performed according to physician practices. In 2253 of 9641 patients (23.4%), PCI was performed by 30 days. Early (<72 hours) PCI was performed in 1228 (12.7%). In 34 placebo patients (5.5%) and 10 treated with eptifibatide (1.7%) (P=0.001), MI preceded early PCI. In patients censored for PCI across the 30-day period, there was a significant reduction in the primary composite end point in eptifibatide patients (P=0.035). Eptifibatide reduced 30-day events in patients who had early PCI (11.6% versus 16.7%, P=0.01) and in patients who did not (14.6% versus 15.6%, P=0.23). After adjustment for PCI propensity, there was no evidence that eptifibatide treatment effect differed between patients with or without early PCI (P for interaction=0.634). PCI was not associated with a reduction of the primary composite end point but was associated with a reduced (nonspecified) composite of death or Q-wave MI. This association disappeared after adjustment for propensity for early PCI. CONCLUSIONS: Eptifibatide reduced the composite rates of death or MI in PCI patients and those managed conservatively. PMID- 10683350 TI - Assessment of aortic valve stenosis severity: A new index based on the energy loss concept. AB - BACKGROUND: Fluid energy loss across stenotic aortic valves is influenced by factors other than the valve effective orifice area (EOA). We propose a new index that will provide a more accurate estimate of this energy loss. METHODS AND RESULTS: An experimental model was designed to measure EOA and energy loss in 2 fixed stenoses and 7 bioprosthetic valves for different flow rates and 2 different aortic sizes (25 and 38 mm). The results showed that the relationship between EOA and energy loss is influenced by both flow rate and aortic cross sectional area (A(A)) and that the energy loss is systematically higher (15+/-2%) in the large aorta. The coefficient (EOAxA(A))/(A(A)-EOA) accurately predicted the energy loss in all situations (r(2)=0.98). This coefficient is more closely related to the increase in left ventricular workload than EOA. To account for varying flow rates, the coefficient was indexed for body surface area in a retrospective study of 138 patients with moderate or severe aortic stenosis. The energy loss index measured by Doppler echocardiography was superior to the EOA in predicting the end points, which were defined as death or aortic valve replacement. An energy loss index 9-weeks) Wistar rats, and we identified the effective are a of the neural response. The extent/size, configuration and orientation of the intrinsic optical response area varied from trial-to-trial with the same whisker stimulation. We argue that the trial-to trial variation was due to cortical blood circulation related to the barrel neural activity. Subsequently, interpolating a family of the traces of the optical response area imaged with repeated stimulation for each animal, we extracted a centered circular area from the trial-to-trial response for each animal. Although the trial-to-trial variation decreased gradually with age, the spatial extent of the interpolated response area was consistently about 660 microm in diameter, in agreement with that measured morphologically and/or histochemically. A possible interpretation is that the optically defined area appears to image the actual effective single-barrel response area, as a first approximation. Furthermore, the constancy of the extracted area independent of age suggests that the barrel cortex is, in fact, virtually mature by 2 weeks of age. The extracted area was also nearly independent of the frequency (>/=5 Hz) of whisker movement. PMID- 10683524 TI - A DPDPE-induced enhancement of inward rectifier potassium current via opioid receptor in neuroblastomaxglioma NG108-15 cells. AB - The effect of a delta-selective opioid agonist, DPDPE([D-Pen(2, 5)]-enkephalin), on the inward rectifier potassium current (I(KIR)) of NG108-15 cell was studied by whole cell voltage-clamp technique. It was found that microM DPDPE increased the amplitude and delayed the activation and inactivation of I(KIR) rapidly and reversibly. These effects could be reversed by naloxone, but were still obtained in pertussis toxin (PTX) preincubated cells or when using GDP-betas (guanosine 5' o-[2-thio] diphoaphate) containing electrodes to block the G-protein coupled events. The above results suggest that DPDPE-induced change of I(KIR) is mediated by delta-opioid receptor but does not involve G-proteins. PMID- 10683525 TI - Characteristics of monocarboxylates as energy substrates other than glucose in rat brain slices and the effect of selective glial poisoning - a 31P NMR study. AB - In rat brain slices we examined the differences in the levels of high-energy phosphates in the presence of various energy substrates by using 31P NMR with a time resolution of 4 min at 25 degrees C. In parallel experiments we recorded population excitatory postsynaptic potentials (EPSPs) from granule cells in rat hippocampal slices. During high K(+) stimulation (8 min) phosphocreatine (PCr) decreased to a low level and recovered to the control level in standard artificial cerebrospinal fluid (ACSF) in about 10 min. Population EPSPs disappeared following high-K(+) stimulation and recovered in standard ACSF. In iodoacetic acid (IAA)-pretreated slices, whereas glucose was unable to support energy metabolism, the PCr level, which decreased following high-K(+) stimulation, recovered in ACSF containing lactate or pyruvate. The half-time of recovery of PCr levels in ACSF containing lactate was longer than that containing glucose. Population EPSPs in standard ACSF were maintained for more than 1 h, but those in ACSF containing lactate decreased gradually by about half in 40 min. In IAA-pretreated slices, when further treated with fluorocitrate (100 microM) for 2 h, the recovery of the PCr level in ACSF containing lactate after high-K(+) stimulation was completely abolished, whereas the recovery of the PCr level in ACSF containing pyruvate was unaffected. These results indicate that neurons can utilize pyruvate as well as glucose, but not lactate, as exogenous energy substrates, and that lactate may be metabolized to pyruvate in glial cells and transported to neurons to be utilized as an energy substrate. PMID- 10683526 TI - The effects of binocular suture and dark rearing on the induction of c-fos protein in the rat visual cortex during and after the critical period. AB - It has been demonstrated in kittens that binocular lid suture has more deleterious and irreversible effects on plasticity of the developing visual system than rearing in complete darkness. The present study using immunocytochemistry focuses on the effects of the two types of visual deprivation on the inducibility of c-fos protein in visual cortical neurons of rats. Rats were subjected to binocular suture or dark rearing for 1 week during (postnatal days 14-21; P14-P21) and after (P50-P57) the critical period for activity dependent modifiability of cortical ocular dominance. In rats of both age groups reared in the normal light-dark condition, only a small number of Fos immunoreactive neurons was obtained in the visual cortex. By contrast, in dark reared pups and adult rats, numerous c-fos neurons were detected in the layers II IV and VI of the visual cortex following a brief light exposure (1 h). In rats of both ages subjected to binocular suture, Fos neurons were detected in the same layers as in the dark-reared rats, but significantly less in number. We speculate that the reduced plasticity of the visual cortex in the rats subjected to binocular suture may be due partly to the repressed AP-1 activity in visual cortical neurons. No significant difference was detected in c-fos expression in the visual cortex between visually manipulated pups and adult rats. PMID- 10683527 TI - The role of fixation point and subjects' readiness in the occurrence of express saccades as revealed by the self-initiation paradigm. AB - The role of fixation and the subjects' response preparedness in producing express saccades were explored in seven human subjects. The occurrence frequencies of the express saccades were compared in the overlap (continuous presentation of fixation point), gap (fixation point offset 0-400 ms prior to target onset) and no-fixation tasks under the conventional and self-initiation paradigms. In the latter paradigm, the subjects, when ready, touched a sensor in order to ignite the target lamp with a delay time of 0-400 ms (target onset delay time). Therefore, the subjects' response preparedness might be expected to be higher than that in the normal paradigm and equated in each subject at the time when the subjects touched a sensor regardless of the paradigms. Although express saccades were produced neither in the normal overlap nor in the normal no-fixation tasks, they could be produced at the rate of 24 and 48% in the overlap and no-fixation tasks under the self-initiation paradigm, respectively. The highest occurrence frequency of express saccades was obtained when the gap paradigm was combined with the self-initiation paradigm with a delay time of 100 ms (62%). The value was higher by 20% than in the normal gap task. At a target onset delay time of 0 ms under the self-initiation paradigm, the occurrence frequency of express saccades was higher in the overlap task than in the gap task. These results suggest that the subjects' response preparedness has a potentiality to produce express saccades without fixation point offset and that fixation point offset at the same time of the target stimulus onset has an interference, rather than facilitatory, influence on the generation of express saccades. PMID- 10683528 TI - Circadian release of excitatory amino acids in the suprachiasmatic nucleus culture is Ca(2+)-independent. AB - We have previously reported that spontaneous release of excitatory amino acids (aspartate and glutamate) show remarkable circadian rhythms in the organotypic slice culture of rat suprachiasmatic nucleus (SCN). Here we showed effects of extracellular Ca(2+) removal and of L-trans-pyrrolidine-2,4-dicarboxylic acid, a glutamate/aspartate uptake inhibitor on the circadian release of excitatory amino acids in the SCN culture. Amino acids were measured by high-performance-liquid chromatography. Removal of extracellular Ca(2+) exerted no effect on the spontaneous release of the excitatory amino acids, while it blocked high K(+) evoked release of the amino acids. Neither the period nor the amplitude of the spontaneous circadian release of amino acids in Ca(2+)-free medium was different from those in the Ca(2+)-containing medium. On the other hand, L-trans pyrrolidine-2,4-dicarboxylic acid increased the excitatory amino acid levels without affecting the amplitude of excitatory amino acid rhythms. These results indicated that the circadian release of excitatory amino acids in the SCN is Ca(2+)-independent and L-trans-pyrrolidine-2,4-dicarboxylic acid- insensitive. Therefore, Ca(2+)-dependent chemical synaptic transmission may not be involved in the circadian rhythm generation in the SCN. PMID- 10683530 TI - Synaptic architecture of glomeruli in lamina II of the chicken spinal cord, as revealed using ultrathin section and freeze fracture techniques. PMID- 10683529 TI - The p44/42 mitogen-activated protein kinase cascade is involved in the induction and maintenance of astrocyte stellation mediated by protein kinase C. AB - The mitogen-activated protein kinase (MAPK) is known to be involved in the differentiation of various types of cells. To understand the role of p44/42 MAPK (ERK1/2) in astrocyte differentiation, we investigated the effects of U0126 and PD98059, specific inhibitors of the MAPK-activating enzyme MEK, on astrocyte morphology in culture. Cultured rat cortical astrocytes exhibited flattened, polygonal morphology in the absence of stimulation, but differentiated into process-bearing stellate cells in response to the membrane-permeable cyclic AMP analog dibutyryl cyclic AMP (dBcAMP) or the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA). dBcAMP-induced astrocyte stellation was not affected by MEK inhibitors, while PMA-induced astrocyte stellation was significantly blocked by U0126 (0.1-10 microM) and PD98059 (10-30 microM). Western blot analysis with an antibody specific for phosphorylated ERK1/2 revealed that PMA, but not dBcAMP, induced phosphorylation of ERK1/2 in a time- and concentration-dependent manner. The PMA-induced astrocyte stellation and ERK1/2 phosphorylation were blocked by specific PKC inhibitors, GF-109203X (0.01 1 microM) and calphostin C (1 microM). In addition, when U0126 or PD98059 was added after treatment with PMA, stellate astrocytes returned to polygonal. These results suggest that the MEK/ERK cascade is involved in the induction and maintenance of astrocyte stellation mediated by PKC, but not by cyclic AMP signaling. PMID- 10683531 TI - Preface PMID- 10683532 TI - Introduction and official welcome to the congress PMID- 10683533 TI - Who needs vaccination against hepatitis viruses? PMID- 10683534 TI - Keynote speech: health education and promotion programmes--the accountability aspect. AB - In the early days of health education, people depended on health education specialists to provide information about diseases and their prevention. The differentiation between health promotion and health education, and the emphasis on the settings approach have resulted in the inclusion of a wide range of people and professions into these activities. This has raised the question of the protection of consumers, and the issue of accountability has become a major focus. This paper briefly traces the history of health education and promotion, with particular emphasis on the way people have dealt with accountability in the past as a platform for the future. The Polish model of hepatitis B vaccination is used to illustrate these developments. PMID- 10683535 TI - Special address: safety of hepatitis B vaccination. PMID- 10683536 TI - Hepatitis B epidemiology in Poland, Central and Eastern Europe and the newly independent states. AB - The incidence of hepatitis B virus (HBV) infection varies considerably in countries in Central and Eastern Europe and the Newly Independent States, but data are difficult to compare between countries because of the large differences in levels of diagnosis, particularly serological identification, and levels of notification. Poland has high levels of diagnosis, including laboratory diagnosis. In the past, the incidence of hepatitis B in Poland was approx. 45 reported cases per 100,000 population, but following the introduction of improved sterilization of medical equipment in 1986 and a selective programme of vaccination in 1991, the incidence fell to about 35 per 100,000 by 1993. In 1993, an intensive vaccination campaign was launched, which has reduced the incidence to under 15 per 100,000. The incidence of HBV infection has decreased across all age groups and in both men and women, and in the under 3 years age group only 32 cases in total were reported in 1997. In 1996 and 1997, there was a slight relative increase in the incidence of HBV infection in men aged 20-24 years. This group may be a target for future vaccination programmes and other activities of control for the infection. PMID- 10683537 TI - Hepatitis B epidemiology in Latin America. AB - The available data on prevalence of hepatitis B virus (HBV) infection in Latin America are incomplete and largely based on analysis of blood banks, which are not stratified by age or social class. An epidemiological study was recently undertaken in six countries in Latin America to update the data. The highest seroprevalence of antibody to the HBV core antigen (anti-HBc) was found in the Dominican Republic (21.4%), followed by Brazil (7.9%), Venezuela (3.2%) and Argentina (2.1%). The lowest seroprevalence was found in Mexico (1.4%) and Chile (0.6%). The seroprevalence in different regions of Brazil varied from 21% in Manaus to 1.2% in Fortaleza. There were no differences in seroprevalence of anti HBc between females and males except in Brazil (significantly higher in males) and in the Dominican Republic (significantly higher in females). In Brazil alone, higher seroprevalence was associated with lower socioeconomic class. In both the Dominican Republic and Brazil, seroprevalence was high in childhood, and in Brazil and Argentina, anti-HBc was detected in 3.0-6.6% of children up to 1 year old, suggesting vertical transmission. Other risk factors included dental and surgical procedures, sexual activity and tattooing. There was an increase in seroprevalence in all countries at or after adolescence, suggesting that sexual activity is a major route of transmission. PMID- 10683538 TI - Hepatitis B epidemiology in Asia, the Middle East and Africa. AB - Asia and Africa have previously been classified as areas of high endemicity for hepatitis B virus (HBV), but in some countries highly effective vaccination programmes have shifted this pattern towards intermediate or low endemicity. Thus, China is now the only country in Asia where HBV endemicity is high. Countries with intermediate endemicity include India, Korea, the Philippines, Taiwan and Thailand, and those with low endemicity include Japan, Pakistan, Bangladesh, Singapore, Sri Lanka and Malaysia. Most countries in Africa have high HBV endemicity, with the exceptions of Tunisia and Morocco, which have intermediate endemicity. Zambia has borderline intermediate/high endemicity. In the Middle East, Bahrain, Iran, Israel and Kuwait are areas of low endemicity, Cyprus, Iraq and the United Arab Emirates have intermediate endemicity, and Egypt, Jordan, Oman, Palestine, Yemen and Saudi Arabia have high endemicity. All of these Middle East countries reach a large proportion of their population with hepatitis B vaccination, which is reducing the infection rate, particularly in Saudi Arabia. The vaccination programme in Taiwan has also greatly reduced the HBV infection rate. Future vaccination programmes must take into account the mode of transmission of HBV, the healthcare infrastructure to deliver vaccination, and the socioeconomic and political factors in each individual country, to determine the most cost-effective way of infection control. PMID- 10683539 TI - Clinical course and consequences of hepatitis B infection. AB - Hepatitis B virus (HBV) is a small enveloped virus containing partially double stranded DNA. The DNA and HBV-specific DNA polymerase are surrounded by the HBV core antigen (HBcAg), which in turn is surrounded by a lipoprotein envelope containing the HBV surface antigen (HBsAg). Serum of HBV-infected patients contains complete virus particles, as well as non-infectious spherical or filamentous HBsAg particles. Acute hepatitis is characterized by the appearance of serum HBV markers, including HBsAg and IgM anti-HBc, which then disappear during convalescence. Persistence of HBsAg for more than 6 months indicates a carrier state. Chronic hepatitis develops in 90% of newborns who become infected, compared with 29-40% of children infected and 5-10% of adults infected. The immune status of the infected person also influences the development of chronic hepatitis. Chronic HBV infection can be diagnosed by serology (identification of HBsAg and HBV DNA), biochemistry (elevated aminotransferase levels) and liver biopsy. The last is important to assess the severity of disease, its stage and prognosis, and to exclude other hepatic diseases. The outcome of chronic HBV infection varies between individuals, with estimated 5-year survivals of 97% for chronic persistent hepatitis, 86% for chronic active hepatitis, and 55% for chronic active hepatitis with cirrhosis. Treatment with interferon alpha is effective in up to 40% of cases, but in view of the very large number of infected people worldwide, vaccination to prevent spread of the disease is a more cost effective option. PMID- 10683540 TI - Teenagers' lifestyle and the risk of exposure to hepatitis B virus. AB - Prevention of lifestyle-related diseases and promotion of physical and mental well-being in adolescents require an understanding of how life situations place adolescents at risk. The most important risk factors for hepatitis B virus (HBV) infection are sexual activity with more than one partner and injecting drug use. Sexual transmission is particularly important in areas of low endemicity, but is increasingly important in areas of high endemicity as young people adopt a 'Western' lifestyle. HBV infection in general is associated with indicators of sexual activity, e.g. number of sexual partners, years of sexual activity and the occurrence of other sexually transmitted diseases (STDs). Risk behaviours are often associated: adolescents who have frequent sexual intercourse also drink more alcohol, smoke more cigarettes, use marijuana more often and wear seat-belts less often when driving than adolescents who have little or no sexual activity. Health education and health promotion are important activities aimed at preventing HBV infection, but the major strategy should be providing immunity from infection before risk-taking behaviour. As for other STDs, this is best achieved by universal vaccination of young adolescents or infants or both. PMID- 10683541 TI - Rationale for the infant and adolescent vaccination programmes in Italy. AB - In Italy in the 1980s, the incidence of acute hepatitis B was about 13 per 100,000, corresponding on average to 7500 new symptomatic cases per year was about 3%, making Italy an area of intermediate endemicity. HBV infection was also associated with 12 per 100,000 deaths from cirrhosis and with 5.1 per 100,000 deaths from hepatocellular carcinoma. In view of the large numbers of pregnant women who were hepatitis B surface antigen (HBsAg)-positive, selective hepatitis B vaccination of all newborns to these mothers and of other high-risk groups was introduced in 1983. Compliance was high among the newborns but low in other high risk groups. Hepatitis vaccination was adopted in Italy in 1991, including each year all newborns, all adolescents aged 12 years and other high-risk groups. Compliance has been nearly 95% for newborns and 80% for adolescents. Since the introduction of vaccination, both the incidence of acute hepatitis B and the prevalence of HBV carriage have fallen, the latter from 3.4% in 1985 to 0.9% in 1996. There is good evidence that these decreases are mainly the result of the vaccination programmes. Although the full economic impact cannot yet be assessed, about 18,000 cases of acute HBV infection have been prevented over the 6 years since starting the mass vaccination programme, with cost savings of about US$ 244,308,000. PMID- 10683542 TI - Nationwide vaccination: a success story in Taiwan. AB - In the early 1980s, 15-20% of the population of Taiwan were estimated to be hepatitis B virus (HBV) carriers. A programme of mass vaccination against hepatitis B was therefore launched in 1984. In the first 2 years, newborns of all HBVsurface antigen (HBsAg)-positive mothers were vaccinated. Since 1986, all newborns, and then year by year pre-school children, primary school children, adolescents, young adults and others have also been vaccinated. Vaccination coverage is over 90% for newborns, with 79% of pregnant women screened for HBsAg. The proportion of babies born to highly infectious carrier mothers who also became carriers decreased from 86-96% to 12-14%; the decrease was from 10-12% to 3-4% for babies of less infectious mothers. Between 1989 and 1993, the prevalence of HBsAg in children aged 6 years also fell from 10.5 to 1.7%. The average annual incidence of hepatocellular carcinoma in children aged 6-14 years decreased significantly from 0.7 per 100,000 in 1981-1986 to 0.36 per 100,000 in 1990-1994 (P<0.01). Similarly, the annual incidence of hepatocellular carcinoma in children aged 6-9 years declined from 0.52 per 100,000 for those born in 1974-1984 to 0.13 per 100,000 for those born in 1986-1988 (P<0.001). The mass vaccination programme is highly effective in controlling chronic HBV infection and in preventing liver cancer in Taiwan. If a coverage rate of 90% of all newborns vaccinated against hepatitis B can be maintained, by the year 2010 the carrier rate in Taiwan is expected to decline to <0.1%. PMID- 10683543 TI - Discussion 1 PMID- 10683544 TI - The expanded programme on immunization calendar in Poland. AB - Poland has a long history of prophylactic vaccination against infectious diseases. Hepatitis B vaccination was introduced in Poland between 1989 and 1996 as part of the Expanded Programme on Immunization (EPI). All newborns and those at high risk of hepatitis B virus (HBV) infection currently receive hepatitis B vaccine free of charge. For many years Poland has reached or exceeded the indicators required by the World Health Organization for vaccination programmes, and about 10% of the population has now been vaccinated against hepatitis B. The incidence of hepatitis B has decreased from about 40 per 100,000 in the early 1990s to 12.7 per 100,000 in 1997. It is hoped to modify the EPI in the future to improve vaccination against mumps, rubella and poliomyelitis. The possible benefit of vaccination against Haemophilus influenzae type b is currently being evaluated. Financial constraints, however, mean that not all of the approved vaccinations can be implemented. The EPI is supported by recommended vaccinations in certain groups, who pay for the vaccines. For hepatitis B, these include children, teenagers, those between 20 and 40 years of age, and those at high risk because of lifestyle or occupation. PMID- 10683545 TI - The hepatitis B prevention education programme in Poland. AB - The main objective of the hepatitis B prevention education programme in Poland is to promote education in the school setting. The programme stems from the national policy for the prevention of hepatitis B virus (HBV) infection and is an element of the national Health Prevention Programme. The main aims of the programme include reducing morbidity from HBV infection by increasing community awareness, facilitating access to vaccination, establishing local lobbies to support the programme, and encouraging cooperation with vaccine producers. The education programme has been implemented in three phases, starting with a pilot programme in 1996 that was extended to half of Poland in 1997 and to the whole country in 1998. The programme is divided into five stages, consisting of meetings at central, voivodship and local levels, vaccination of children and evaluation of the programme. PMID- 10683546 TI - Evaluation of the hepatitis B prevention education programme in Poland. AB - Evaluation of the hepatitis B prevention education programme was performed as a survey including all the coordinators in 25 regions (211 coordinators; half the country). The success of the programme was defined by an objective measure (the ratio of vaccinated children to the total number of children in a region) and a subjective measure (the need to introduce changes in current procedures). The best information was felt to be provided on the subjective area, with financial aspects raising the most doubts. There was a high level of participation in local training by school nurses, but insufficient participation by paediatric nurses and paediatricians. In one-third of the regions schools fulfilled the tasks set them, but in 19% of the regions only a few schools did. Information about vaccination was given in most public children's clinics in 65% of the regions, but there was no activity in 6% of the regions. Recruitment of sponsorship was successful, with only 12% of parents paying the full cost of vaccination. The most important factors contributing to the success of the education programme were the health education and the epidemiology departments of state sanitary inspection, the schools' medical services and paediatric services. The greatest perceived need for change in procedures was in the area of cooperation with local authorities. Only 56% of those surveyed felt the director of the local sanitary station participated to a significant extent, though the programme is very dependent on such participation. Overall, 12% of the regions achieved a ratio of 20 or more between vaccinated and non-vaccinated children, with another 52% reaching a ratio of 5-19.9. The perceived need for changes in cooperation with the Health Service was smaller in regions in which the ratio of vaccinated children was higher. The survey showed a connection between the results of the education programme and the incidence of hepatitis B virus infection. PMID- 10683547 TI - Cost-benefits of vaccination programmes. AB - Decision-makers are increasingly demanding hard economic data as a basis for the allocation of limited healthcare resources. The main types of evaluation that are available are cost-benefit analysis, cost-effectiveness analysis and cost-utility analysis. Cost-effectiveness analysis is a tool that helps decision-makers to decide on the best use of allocated resources, whereas cost-benefit analysis is a tool that helps policy-makers decide on the overall allocation of resources. The basis of the cost-utility analysis is the quality-adjusted life year (QALY), which allows a direct comparison of a wide range of medical interventions. The cost per QALY for a range of childhood vaccinations can be compared in order to plan a vaccination programme. Public health vaccines warrant a cost-benefit approach, in order to determine if they are worthwhile, whereas recommended vaccines might be more usefully assessed by cost-effectiveness analysis. It is also important to look at combinations of vaccines, which offer large economic advantages by reducing costs (time and equipment) and ensuring better acceptance (improved coverage and reduced risk of disease). Although cost-savings do not necessarily equate with cost-effectiveness, cost-savings are achieved in many vaccination programmes. PMID- 10683549 TI - Discussion 2 PMID- 10683548 TI - Cost-benefit analysis of the Polish hepatitis B prevention programme. AB - This paper presents a cost-benefit analysis of hepatitis B vaccination in Poland. The costs relating to hepatitis B are based on official epidemiological data and a validated model, and amount to 1324.2 million Polish zlotys. The annual costs of vaccination amounted to 120 million zlotys, which when projected over a 20 year period with appropriate adjustment give a total cost of the vaccination programme of 2676 million zlotys. If vaccination is held at its present level for the next 20 years, the total value of benefits would be 12,046.4 million zlotys and of losses due to hepatitis B despite vaccination would be 14,437.0 zlotys. If no hepatitis B vaccination were performed, however, the total loss due to HBV infection is estimated to be 26,484.2 million zlotys. There is clearly a wide gap between the losses due to HBV infection despite vaccination and the costs of the vaccination programme. This clearly shows that the sooner total community vaccination is achieved, the greater the benefits. This may be achieved by vaccinating schoolchildren in the first year of school and by broadening the definitions of high-risk groups. PMID- 10683550 TI - Hepatitis A shifting epidemiology in Latin America. AB - In the past, Latin America was considered to be an area of high endemicity for hepatitis A virus (HAV) infection, with most people infected in early childhood. A seroepidemiological study was recently undertaken in six countries to determine whether this pattern has changed. The highest seroprevalence of antibodies to HAV (anti-HAV) was found in Mexico and the Dominican Republic. Analysis of the different age groups showed that at age 6-10 years, 30% of children in Chile and 54-55% in Brazil, Venezuela and Argentina had been infected, compared with almost 70% in Mexico and 80% in the Dominican Republic. At age 11-15 years, nearly 90% in Mexico and 91% in the Dominican Republic had been infected, compared with 54% in Argentina, 62% in Venezuela, 60% in Brazil and 70% in Chile. By age 31-40 years, over 80% of the populations in all six countries had been exposed to HAV. In all of the countries except Brazil and Venezuela, the seroprevalence of anti HAV was significantly higher in females than in males. In Mexico, Argentina and Brazil, anti-HAV seroprevalence was significantly higher in the low socioeconomic groups than in the middle/high socioeconomic groups. The results show that there has been a shift from high to medium endemicity of HAV infection throughout Latin America, which may result in more clinical cases in adolescents and adults and a greater potential for outbreaks. The vaccination strategy for hepatitis A should thus be reviewed. PMID- 10683551 TI - Hepatitis A shifting epidemiology in South-East Asia and China. AB - A review of the epidemiology of hepatitis A virus (HAV) infection over the last 20 years shows shifting patterns in the prevalence of antibodies to HAV (anti HAV) throughout South-East Asia and China. A number of countries have shifted from high to moderate and from moderate to low endemicity, with a corresponding increase in the age of exposure from childhood to early adulthood. The changes have resulted from improvements in hygiene, sanitation and the quality of drinking water, reflecting improvements in living standards and socioeconomic progress. In general in the late 1970s and early 1980s, 85-95% of the population of developing countries like the Philippines, Korea, China and Thailand were anti HAV-positive by age 10-15 years, compared with only about 50% in the more affluent countries like Malaysia and Singapore. In the early 1990s, 85-95% of the population were immune by age 30-40 years in the Philippines, Korea, China and Thailand, and by 50 years of age and above in Malaysia and Singapore. Similar trends were noted in Hong Kong, Taiwan and Japan. Exposure to HAV at a later age may be associated with an increase in hepatitis A morbidity and a greater propensity for outbreaks. PMID- 10683552 TI - Hepatitis A shifting epidemiology in the Middle East and Africa. AB - Data on the endemicity of hepatitis A virus (HAV) infection in Africa and the Middle East are scant, but most of Africa appears to remain a high endemicity region, with the exception of subpopulations in some areas, e.g. White people in South Africa. Saudi Arabia is a model for the Middle East, and is a country in which shifting HAV epidemiology has been documented in recent years, concurrent with the social and economic development that has occurred over the last two decades. Earlier studies generally showed very high prevalence rates, with most people becoming infected in early childhood. Between 1989 and 1995, however, there was a significant fall in the seroprevalence of antibodies to HAV in children up to 12 years old throughout the country except in one region bordering the Yemen. The highest seroprevalence is found in children from rural backgrounds, while the seroprevalences in Bedouin and urban children are similar. Seroprevalence is related to socioeconomic status, being highest in the lowest groups. Similar findings have been reported from other countries in the Middle East. The existence of pockets of high endemicity for HAV infection with surrounding areas shifting towards intermediate endemicity may lead to outbreaks, and widespread vaccination should be considered. PMID- 10683553 TI - Hepatitis A shifting epidemiology in Poland and Eastern Europe. AB - The clinical morbidity of hepatitis A probably only represents 20% of cases of hepatitis A virus (HAV) infection. When it became possible to determine specific antibodies, a seroepidemiological survey of anti-HAV was undertaken in Poland, which showed that between 1979 and 1997 there was a shift in the peak age of infection from childhood to adulthood, concomitant with a substantial decline in the incidence of HAV infection. Data from the World Health Organization also indicate that there has also been a decline in the incidence of hepatitis A in Eastern European countries in general, over the 3 years from 1994 to 1996. The potential risk of epidemics still exists, however, when appropriate conditions are created. The available data show that fewer young people are becoming infected with HAV, and general preventive measures, including vaccination of children and high-risk groups (e.g. healthcare and childcare personnel and those living in 'closed communities') are needed to deal with HAV infections in Eastern Europe. PMID- 10683555 TI - Discussion 3 PMID- 10683554 TI - Clinical course and consequences of hepatitis A infection. AB - Hepatitis A virus (HAV) is a small, non-enveloped RNA virus belonging to the Picornaviridae, for which only one serotype has been identified. Transmission is usually through the faecal-oral route by person-to-person contact. The most common risk factors are household or sexual contact with a sufferer, attendance or working at a day-care centre, international travel, and association with food or waterborne outbreaks; 55% of cases have no identifiable risk factors. HAV infection may be symptomatic or asymptomatic, and shows three phases. Virus is shed during the incubation phase, anti-HAV IgM appears during the symptomatic phase and can be used for diagnosis, and anti-HAV IgG appears at the same time but persists lifelong. Unusual clinical manifestations of hepatitis A include cholestatic, relapsing and fulminant hepatitis. Hepatitis A accounts for 93% of cases of acute hepatitis in Argentina, including 7% of atypical clinical cases. Hepatitis A is the major cause of fulminant hepatitis, and has been reported to account for 10% of liver transplants in children in France and 20% in Argentina. One-year survival after liver transplantation is 64%. Prevention must be considered as the main means of averting this severe illness. PMID- 10683556 TI - Hepatitis B school-based vaccination programmes in the USA: a model for hepatitis A and B. AB - Following the recommendation for routine vaccination against hepatitis B virus for newborns, many states have started school-based catch-up vaccination of 11- to 12-year-olds. Implementation of these programmes requires educational and promotional initiatives to increase awareness among parents, children, teachers, school nurses, school boards and administration. Experience in Framingham, Massachusetts, suggests that over 90% of targeted hepatitis B vaccine coverage can be achieved. Because hepatitis B vaccination targeted at high-risk groups in the USA was largely unsuccessful, this suggests that the initial similar targeted approach with hepatitis A vaccination will also fail. Only about 50% of hepatitis A cases have a known risk factor, and multiple high-risk areas exist throughout the USA. However, the geographical clustering of these high-risk areas and the occurrence of periodic outbreaks, suggest that school-based hepatitis A vaccination programmes may be effective in reducing the risk of infection. A voluntary programme in San Antonio achieved 43% of the targeted coverage in its first year, and a compulsory programme is due to start in Oklahoma. The effectiveness of this programme is not yet known, but future recommendations are likely to include hepatitis A vaccination as a school entry requirement in areas with high incidence of hepatitis A. PMID- 10683557 TI - Prospects for vaccination against hepatitis A and B in Catalonia (Spain). AB - Catalonia is in an area of intermediate endemicity for hepatitis A virus (HAV) infection. An Expert Committee has recently proposed the implementation of universal hepatitis A vaccination for 12-year-olds, based on the fact that no risk factors can be identified for hepatitis A in 50% of cases, and also that selective vaccination targeted at high-risk groups has a limited potential to reduce the incidence of hepatitis A. The well-established programme of hepatitis B vaccination of pre-adolescents in Catalonian schools has high levels of vaccination coverage. This will provide a means to introduce hepatitis A vaccination in a cost-effective way in schools, by replacing the single vaccine with the combined hepatitis A and B vaccine. High-risk groups will also continue to be targeted. A pilot programme has commenced in the 1998/1999 school year and will be evaluated after 3 years. If it is successful, it will be extended indefinitely. PMID- 10683558 TI - From hepatitis B to hepatitis A and B prevention: the Puglia (Italy) experience. AB - The incidence of hepatitis B virus infection in Italy is 10 per 100, 000 population, with most cases occurring in young adults. Vaccination against hepatitis B has been compulsory since 1991 for all newborns and 12-year-olds. In the Puglia region, this programme has reduced the incidence of hepatitis B from 7.4 per 100,000 population in 1990 to 2.4 per 100,000 population in 1996. The number of notified cases of hepatitis B in Puglia decreased from 212 in 1992 to 73 in 1997. As 50% of these cases occurred in young adults, the main aim of the current vaccination programme is to achieve high coverage rates among teenagers and young adults within the next few years. Although the incidence of hepatitis A is only about 5 per 100, 000 overall in Italy, Puglia is an area of intermediate endemicity with a seroprevalence of antibodies to hepatitis A virus (anti-HAV) of about 40% in 18-year-olds. The incidence of hepatitis A is up to 30 per 100,000 between the periodic outbreaks that occur every 2-4 years. Most notified cases occur in adolescents and young adults. The last outbreak of about 11,000 cases of hepatitis A in the Puglia region occurred in 1996-1997, mainly in the summer months in towns with harbours or near the coast. The most important risk factor was initially consumption of raw seafood, but later was personal contact, probably between children. A vaccination programme against hepatitis A was initiated in Puglia in 1997, aiming to vaccinate all infants of 15-18 months and all 12-year-olds against hepatitis A. Infants receive monovalent hepatitis A vaccine with the first dose of mumps/measles/rubella vaccine. Monovalent hepatitis vaccine can be given with the second and third doses of hepatitis B vaccine in 12-year-olds, but use of combined hepatitis A and B vaccine is recommended to aid compliance and reduce the commitment of physician/nurse time. Vaccination can be performed in school. PMID- 10683559 TI - Cost-effectiveness of hepatitis A and B vaccination programme in Germany. AB - A recent study in Germany analysed the epidemiological and economic impact of combined hepatitis A and hepatitis B vaccination of all 1-15-year-olds or of all 11-15-year-olds projected over three periods of 10 years compared with a strategy of non-vaccination. Vaccination of all 1-15-year-olds will achieve a reduction of 57,596 new hepatitis A cases with 7555 new infections remaining over 30 years. Vaccination of all 11-15-year-olds will reduce the number of new hepatitis A cases by 19,826, with 45,325 new cases remaining over 30 years. Vaccination of all 1-15-year-olds will reduce the number of new hepatitis B cases by 45,820 with 7484 new cases remaining over 30 years, compared with vaccination of all 11-15 year-olds, which will reduce the number of new hepatitis B cases by 21,905 with 31,339 new cases remaining over 30 years. This significant reduction in the number of new cases will lead to savings in treatment costs of DM 2.9 billion for vaccination of 11-15-year-olds and of DM 5.1 billion for vaccination of 1-15-year olds. The cost-effectiveness of vaccination ranges from costs of DM 90,000 for each infection avoided to savings of DM 50, 000 for each case avoided. If unreported cases are also taken into account and are equally distributed between all age-groups, the savings per infection avoided over 30 years are DM 69,796 for vaccination of 11-15-year-olds and DM 55,850 for vaccination of 1-15-year-olds. Although the strategy of vaccinating 11-15-year-olds is the more cost-effective, it leaves a large percentage of the population at high risk of hepatitis A virus infection. The use of an initially more expensive combined hepatitis A and B vaccine represents a cost-effective alternative to monovalent hepatitis B vaccination and is more beneficial in terms of its epidemiological impact. PMID- 10683561 TI - Discussion 4 PMID- 10683560 TI - Awareness campaigns: experience in Mexico. AB - The current total of AIDS cases in Mexico is 37,000 of which 86% have occurred in men. The major route of transmission is sexual. The campaign to prevent AIDS has fallen into four phases, and has now been extended to other sexually transmitted diseases, including hepatitis B. The first phase (1985-1989) was based around question and answer brochures, which increased awareness but did not remove misconceptions. A mass media campaign addressed these misconceptions and stressed preventive measures. The campaign was halted by opposition to the promotion of condom use on the grounds that it encouraged promiscuity. The second phase (1989 1992) used more conservative messages, but these were too obscure and failed to reach the target audience. A poster campaign using popular lottery characters was widely accepted. In the third phase (1992-1994), a combination of messages was targeted at different populations, including parents and women, and general public sympathy for social support for people with AIDS was encouraged. In the fourth phase (1996-2000), a mass media campaign was aimed at teenagers, with parents and teachers as support groups. The campaign was widened to include HBV infection, and posters and brochures for teenagers were produced. These are distributed as part of a collaboration with non-governmental organizations providing sex education. The private medical sector is being encouraged to provide facilities for hepatitis B vaccination. So far the campaign has only been established in Mexico City, but it is hoped that this will be extended nationwide. Hepatitis B vaccination has been recently included in the National Immunization Programme for infants in the first year of life and it is officially recommended for at-risk populations. PMID- 10683563 TI - Discussion of the regional workshop conclusions PMID- 10683562 TI - Introduction to the regional workshops. PMID- 10683564 TI - P2 receptor-mediated inhibition of dopamine release in rat neostriatum. AB - Axon terminal nucleotide P2 receptors mediating an inhibition of transmitter release have, so far, been detected in various sympathetically innervated tissues,(8,27) and on central noradrenergic,(14,26) glutamatergic(15) and serotonergic neurons. (28) We have now investigated the effect of ATP and related nucleotides on the release of endogenous dopamine from slices of rat neostriatum using fast cyclic voltammetry. Mutual interactions between the two neurotransmitters have been observed previously: ATP and related nucleotides induce a release of dopamine in PC12 pheochromocytoma cells, a frequently used model for sympathetic neurons;(10,22) they also increase the dopamine concentration in rat brain measured by in vivo microdialysis(16,32) and stimulate the uptake of dopamine by rat striatal synaptosomes.(3) Dopamine, in contrast, facilitates activation of ligand-gated cation channels (i. e. P2X(2) receptors) by ATP.(11,20) Here, we show that ATP and two of its analogues decrease the electrically evoked release of endogenous dopamine in rat neostriatum. The inhibitory effect of ATP is blocked by the P2 receptor antagonists suramin, reactive blue 2 and cibacron blue 3GA. Suramin, in addition, partly prevents the attenuation of dopamine release evoked by a single stimulus that follows a brief train of high-frequency pulses.These findings suggest the existence of release inhibiting P2 receptors on dopaminergic nerve terminals and indicate that dopaminergic transmission in rat neostriatum might be modulated by an endogenous P2 receptor ligand, presumably ATP. PMID- 10683565 TI - Ceramide-induced sustained depression of synaptic currents mediated by ionotropic glutamate receptors in the hippocampus: an essential role of postsynaptic protein phosphatases. AB - Ceramide, a sphingomyelin-derived second messenger, mediates cellular signals of cytokines such as tumor necrosis factor-alpha that are rapidly produced in the brain in response to vigorous neuronal activity and tissue injury. Using whole cell patch-clamp recordings, the present study examined whether ceramide modulated excitatory postsynaptic currents mediated by ionotropic glutamate receptors in CA1 pyramidal neurons of rat hippocampal slices. Application of N acetyl-D-sphingosine, a synthetic cell-permeable ceramide analog, promptly produced a slight increase of excitatory postsynaptic current amplitude lasting for about 3 min. However, this transient enhancement was followed by a profoundly delayed-onset, sustained depression of synaptic excitatory postsynaptic currents in a concentration-dependent fashion (1-30 microM). This ceramide-induced sustained depression was not associated with changes in paired-pulse facilitation, a phenomenon resulting from an alteration of presynaptic transmitter release. Dihydro-N-acetyl-D-erythro-sphingosine (10 microM), an inactive analog of N-acetyl-D-sphingosine, did not affect synaptic excitatory postsynaptic currents, indicating the specificity of N-acetyl-D-sphingosine's action. The induction of ceramide-induced sustained depression was primarily dependent on the activation of postsynaptic protein phosphatases, being considerably blocked by loading 30 nM okadaic acid (a potent inhibitor of protein phosphatases 1 and 2A) into neurons. In addition, following a stable establishment of ceramide-induced sustained depression, a protocol for inducing long-term depression caused no additional decreases in excitatory postsynaptic current amplitude, and vice versa. The study suggests that ceramide induces a long-term depressed modulation on synaptic transmission mediated by ionotropic glutamate receptors in the hippocampus, possibly through the activation of postsynaptic protein phosphatases 1 and 2A. In addition, ceramide-induced sustained depression seems to share some common mechanisms with long-term depression, such as the cascades of events resulting from the activation of protein phosphatases. Collectively, the long-term depressed modulation of ceramide on ionotropic glutamate receptor-mediated functions may be particularly important in various physiological and/or pathological conditions, in which the ceramide signaling pathway is activated in the mammalian brain. PMID- 10683566 TI - Long-term potentiation induction--a synaptic catch mechanism released by extracellular phosphorylation. AB - The best understood form of long-term potentiation in hippocampal CA1 neurons is induced by activation of the N-methyl-D-aspartate receptor complex and subsequent activation of the intracellular second messenger systems. In addition to this intracellular mechanism, long-term potentiation can also be induced by an extracellular mechanism involving phosphorylation by ATP-ecto-protein kinase. In the present study, we hypothesize that a putative blocking molecule of the formation of long-term potentiation exists on the synaptic membrane, and examine whether ecto-protein kinases play a role in the block of long-term potentiation by phosphorylating the ecto-domains of this molecule in CA1 neurons of guinea-pig hippocampal slices. Long-term potentiation was induced by theta burst stimulation whether or not the test input was delivered to the CA1 neurons following burst stimulation. However, 5 microM K-252b, an ecto-protein kinase inhibitor, only blocked the induction of long-term potentiation when the test input was delivered during a 30-min period following burst stimulation. The results suggest that the process of formation of long-term potentiation continues independently of test synaptic input, while the block of long-term potentiation results from a combination of an interruption of the ATP-ecto-protein kinase-dependent processes and continued test synaptic input after burst stimulation. This block of long term potentiation, which should be released by activation of ATP-ecto-protein kinase, is suggested to act as a "safety catch" for synaptic plasticity in hippocampal CA1 neurons. PMID- 10683567 TI - Differential regulation of Ca(2+)-calmodulin stimulated and Ca(2+)-insensitive adenylyl cyclase messenger RNA in intact and denervated mouse hippocampus. AB - The Ca(2+)-calmodulin stimulated AC1 and Ca(2+)-insensitive AC2 are major isoforms of adenylyl cyclase, playing an important role in synaptic plasticity in the mammalian brain. We studied the pattern of expression of AC1 and AC2 genes in the hippocampus of C57BL/6 mice. We found that there were differences in their patterns of distribution in the dentate gyrus. AC1 messenger RNA was detected both in the dentate granule cell bodies and the corresponding molecular field whereas AC2 messenger RNA was preferentially distributed in the dentate granule cell layer, suggesting that AC1 and AC2 messenger RNA are differentially regulated in the dentate gyrus. In order to examine the regulation of AC1 and AC2 expression in response to synaptic deafferentation and reinnervation, the distribution patterns of the two AC messenger RNA in the hippocampal fields and the parietal cortex were analysed 2, 5, 9 and 30 days following an unilateral entorhinal cortex lesion. Interestingly, we found significantly reduced levels of AC1 hybridization signal following the lesion whereas the level of AC2 messenger RNA remained unaffected in all lesioned groups. The changes in AC1 messenger RNA were transient, with a maximal reduction at five days postlesion, and were restricted to the granule cell bodies and stratum moleculare of the deafferented dentate gyrus. No significant change in AC1 messenger RNA levels was detected in other hippocampal fields nor for any other postlesion times studied. These findings suggest that, at least in the dentate gyrus, messenger RNA for AC1 and AC2 might be differentially compartmentalized in cell bodies and dendritic fields. The activity-dependent regulation of AC1 messenger RNA levels by afferent synapses may provide an elegant mechanism for achieving a selective local regulation of AC1 protein, close to its site of action. PMID- 10683568 TI - Immunohistochemical and neurochemical correlates of learning deficits in aged rats. AB - This study examined whether cholinergic and monoaminergic dysfunctions in the brain could be related to spatial learning capabilities in 26-month-old, as compared to three-month-old, Long-Evans female rats. Performances were evaluated in the water maze task and used to constitute subgroups with a cluster analysis statistical procedure. In the first experiment (histological approach), the first cluster contained young rats and aged unimpaired rats, the second one aged rats with moderate impairment and the third one aged rats with severe impairment. Aged rats showed a reduced number of choline acetyltransferase- and p75(NTR)-positive neurons in the nucleus basalis magnocellularis, and choline acetyltransferase positive neurons in the striatum. In the second experiment (neurochemical approach), the three clusters comprised young rats, aged rats with moderate impairment and aged rats with severe impairment. Alterations related to aging consisted of reduced concentration of acetylcholine, norepinephrine and serotonin in the striatum, serotonin in the occipital cortex, dopamine and norepinephrine in the dorsal hippocampus, and norepinephrine in the ventral hippocampus. In the first experiment, there were significant correlations between water maze performance and the number of; (i) choline acetyltransferase- and p75(NTR) positive neurons in the nucleus basalis magnocellularis; (ii) choline acetyltransferase-positive neurons in the striatum and; (iii) p75(NTR)-positive neurons in the medial septum. In the second experiment, water maze performance was correlated with the concentration of; (i) acetylcholine and serotonin in the striatum; (ii) serotonin and norepinephrine in the dorsal hippocampus; (iii) norepinephrine in the frontoparietal cortex and; (iv) with other functional markers such as the 5-hydroxyindoleacetic acid/serotonin ratio in the striatum, 3,4-dihydroxyphenylacetic acid/dopamine ratio in the dorsal hippocampus, 5 hydroxyindoleacetic acid/serotonin and homovanillic acid/dopamine ratios in the frontoparietal cortex, and 3,4-dihydroxyphenylacetic acid/dopamine ratio in the occipital cortex. The results indicate that cognitive deficits related to aging might involve concomitant alterations of various neurochemical systems in several brain regions such as the striatum, the hippocampus or the cortex. It also seems that these alterations occur in a complex way which, in addition to the loss of cholinergic neurons in the basal forebrain, affects dopaminergic, noradrenergic and serotonergic processes. PMID- 10683569 TI - Apolipoprotein E-deficient mice are not more susceptible to the biochemical and memory deficits induced by nucleus basalis lesion. AB - We investigated whether the nucleus basalis lesion induced by quisqualic acid was associated with a more severe impairment of spatial navigation in a water maze, a greater reduction in frontal choline acetyltransferase activity and decrease in the number of choline acetyltransferase-positive neurons in the nucleus basalis in apolipoprotein E-deficient mice than in control mice. We also studied the effect of ageing on water maze spatial navigation and cortical choline acetyltransferase activity in 16-month-old control and apolipoprotein E-deficient mice. We found that the lesion decreased choline acetyltransferase-positive neurons in the nucleus basalis and frontal choline acetyltransferase activity equally in control and apolipoprotein E-deficient mice. The nucleus basalis lesion had no effect on the initial acquisition in the water maze in control and apolipoprotein E-deficient mice after 25 or 106 days of recovery. However, the nucleus basalis lesion impaired the reversal learning in the water maze similarly in both strains after 25 days of recovery, but had no effect after 106 days of recovery. Finally, water maze spatial navigation and cortical choline acetyltransferase activity were similar in old control and apolipoprotein E deficient mice. These results suggest that young and old apolipoprotein E deficient mice do not have impairments in cholinergic activity or spatial navigation. Furthermore, apolipoprotein E deficiency does not increase the sensitivity to cholinergic and spatial navigation deficits induced by lesioning of the nucleus basalis with an excitatory amino acid and does not slow down the behavioral recovery. PMID- 10683570 TI - Muscarinic receptors depress GABAergic synaptic transmission in rat midbrain dopamine neurons. AB - The effects of muscarine and nicotine on evoked and spontaneous release of GABA were studied using intracellular and whole-cell patch-clamp recordings from rat midbrain dopamine neurons in an in vitro slice preparation. Muscarine (30 microM) reversibly depressed the pharmacologically isolated inhibitory postsynaptic potential evoked by local electrical stimulation. The maximal inhibition of the inhibitory postsynaptic potential amplitude was 39.6+/-5%. This depressant effect of muscarine was blocked by the M3/M1 receptor antagonist 4-diphenylacetoxy-N methylpiperidine methiodide (100 nM), but was slightly affected by the M1/M3 receptor antagonist pirenzepine (1 microM). In addition, muscarine decreased the frequency of the miniature synaptic currents without any effect on their amplitude. Moreover, muscarine did not change the GABA-induced hyperpolarization, indicating that its effect on the inhibitory postsynaptic potential is mediated by presynaptic receptors. On the contrary, the cholinergic agonist nicotine did not change the frequency or the amplitude of the spontaneous glutamatergic and GABAergic synaptic currents. Our data indicate that a prevalent activation of presynaptic M3 muscarinic receptors inhibits the GABA-mediated synaptic events, while the activation of nicotinic receptors does not affect the release of glutamate and GABA on midbrain dopamine neurons. PMID- 10683571 TI - Oxidative stress during energy impairment in mesencephalic cultures is not a downstream consequence of a secondary excitotoxicity. AB - Past studies have shown that inhibiting energy metabolism with malonate in mesencephalic cultures damages neurons by mechanisms involving N-methyl-D aspartate receptors and free radicals. Overstimulation of N-methyl-D-aspartate receptors is known to produce free radicals. This study was, therefore, carried out to determine if N-methyl-D-aspartate receptor activation triggered by energy impairment was a significant contributor to the oxidative stress generated during energy inhibition. Exposure of mesencephalic cultures to malonate for the minimal time required to produce toxicity, i.e. 6h, resulted in an increase in the efflux of both oxidized and reduced glutathione, and a decrease in tissue levels of reduced glutathione. In contrast, exposure to 1mM glutamate for 1h caused an increased efflux of reduced glutathione, but no changes in intra- or extracellular oxidized glutathione or intracellular reduced glutathione. Blocking N-methyl-D-aspartate receptors with MK-801 (0.5 microM) during malonate exposure did not modify malonate-induced alterations in glutathione status or free radical generation as monitored by dihydrochlorofluorescein diacetate and dihydrorhodamine 123 fluorescence. In contrast, the increase in dihydrorhodamine fluorescence caused by glutamate was completely blocked by MK-801. Reduction of tissue glutathione with a 24h pretreatment with 10 microM buthionine sulfoxamine, as shown previously, greatly potentiated malonate-induced toxicity to dopamine and GABA neurons, but had no potentiating effect on toxicity due to glutamate. The findings indicate that although oxidative stress mediates damage due either to energy deprivation or excitotoxicity, N-methyl-D-aspartate receptor over stimulation does not contribute significantly to the oxidative stress that is incurred during malonate exposure. PMID- 10683572 TI - Role of climbing fibers in determining the spatial patterns of activation in the cerebellar cortex to peripheral stimulation: an optical imaging study. AB - The spatial patterns of activation in the rat cerebellar cortex evoked by ipsilateral face stimulation were mapped using optical imaging based on the pH sensitive dye, Neutral Red. The aims of the study were to characterize the optical responses evoked by peripheral stimulation and test the hypothesis that the resultant parasagittal banding is due to climbing fiber activation. In the anesthetized rat Crus I and II of the cerebellar cortex were stained with Neutral Red. Epi-fluorescent changes produced by a train of stimuli (5-10s and 4-20 Hz) to the ipsilateral face were monitored in time using a fast, high resolution charge-coupled device camera. The patterns of activation were quantified using a two-dimensional fast Fourier transform analysis that removed signals with high spatial frequencies and minimized the contribution of horizontal structural elements (i.e. blood vessels). The dominant spatial pattern of activation evoked by face stimulation was that of parasagittal bands. The bands were highly frequency-dependent and were elicited most strongly by stimulus frequencies in the range of 6-8 Hz. There was a large fall-off in the response for frequencies above and below. The optical signal evoked by face stimulation built up over a period of 10s and then gradually decayed. Within a folium the individual parasagittal bands exhibited some frequency and temporal specificity. Stimulation of the contralateral inferior olive also resulted in the activation of parasagittal bands with characteristics similar to the bands evoked by face stimulation, including a preferred stimulus frequency which peaked at 10 Hz. Injection of lidocaine into the contralateral inferior olive blocked the parasagittal bands evoked by ipsilateral face stimulation, while control injections of saline had no effect. The results confirm that a parasagittal banding pattern is a dominant feature of the functional architecture of the cerebellar cortex. The parasagittal banding pattern observed with Neutral Red is due primarily to the activation of climbing fiber afferents. The frequency tuning of the responses, with the preference for peripheral stimuli of 6-8 Hz, is in agreement with previous findings that the inferior olive is inherently rhythmic. These observations support the hypothesis that inferior olivary neurons are dynamically coupled into groups that activate parasagittal bands of Purkinje cells in the cerebellar cortex. The frequency tuning also supports the hypothesis that the climbing fiber system is involved with timing. Activation of this afferent system may require stimuli with appropriate frequency content and stimuli synchronized to the rhythmicity of the inferior olive. PMID- 10683573 TI - A direct cerebrocerebellar projection in adult birds and rats. AB - The rostral Wulst of birds, like the somatosensory cortex of mammals, receives somatosensory information from the thalamus and projects to the brainstem and spinal cord via a pyramidal-like tract. Using anterograde and retrograde tract tracers, we show here, in adult zebra finches, that the rostral Wulst also projects directly to the cerebellar cortex and deep nuclei. In the cortex, the cerebrocerebellar fibers resemble neither mossy nor climbing fibers, but more closely resemble the multilayer fibers shown to originate from the hypothalamus in mammals. We also show that a sparse projection to the cerebellum from the mammalian neocortex, originally thought to be lost during early development, is present in the adult rat. Although the functional implications of these results are obscure, they suggest a revision of the concept of the "cerebrocerebellar system", which is generally considered to involve a pontine relay. PMID- 10683574 TI - Apposition of neuronal elements containing nitric oxide synthase and glutamate in the nucleus tractus solitarii of rat: a confocal microscopic analysis. AB - The distribution of glutamate and neuronal nitric oxide synthase in the rat nucleus tractus solitarii was investigated by double fluorescent immunohistochemistry combined with confocal laser scanning microscopy. Cells and fibers that exhibited neuronal nitric oxide synthase immunoreactivity alone, glutamate immunoreactivity alone or both immunolabels were present in all subnuclei of the nucleus tractus solitarii, but staining intensities differed between the subnuclei. The percentages of double-labeled glutamate-immunoreactive cells also differed between the subnuclei. The central subnucleus contained the highest percentage of double-labeled glutamate-immunoreactive cells and the medial subnucleus contained the lowest. The percentages of double-labeled neuronal nitric oxide synthase-immunoreactive neurons likewise differed between the subnuclei. The central subnucleus contained the highest percentage of double labeled neuronal nitric oxide synthase-immunoreactive neurons and the commissural subnucleus contained the lowest. Because of our interest in cardiovascular regulation, the anatomical relationship between glutamate-immunoreactive and neuronal nitric oxide synthase-immunoreactive fibers in the dorsolateral and commissural subnuclei was further examined at higher magnification. Close appositions were observed between neuronal nitric oxide synthase-immunoreactive and glutamate-immunoreactive fibers, between double-labeled and glutamate immunoreactive fibers, and between neuronal nitric oxide synthase-immunoreactive and double-labeled fibers. We recognized that a single visual perspective might cause labeled fibers that pass in close proximity to appear to make contact. Therefore, we constructed three-dimensional images from serial optical sections obtained from the dorsolateral and commissural subnuclei by means of a confocal scanning microscope. Rotation of the three-dimensional images caused some fibers that had seemed to be in close apposition to other structures to separate from those structures. In contrast, some glutamate-immunoreactive and some neuronal nitric oxide synthase-immunoreactive fibers remained in close apposition regardless of the angle at which they were viewed. This study supports there being an anatomical link between glutamatergic and nitroxidergic systems in the nucleus tractus solitarii. Recognized physiological interactions between the two systems could occur through such a link. PMID- 10683575 TI - Fos expression is induced by increased nitric oxide release in rat spinal cord dorsal horn. AB - The relationship between exogenous or endogenous nitric oxide and c-fos, an immediate-early gene which can further activate the production of other substances in the central nervous system, was investigated in this study. We found that Fos expression is increased after intradermal capsaicin injection, which also leads to endogenous nitric oxide release in the spinal cord. The increased Fos expression is distributed in neurons of the superficial layers and lamina V of the dorsal horn on the side ipsilateral to the injection. The increased Fos expression is blocked by N(G)-nitro-L-arginine methyl ester, a nitric oxide synthase inhibitor, but not by its inactive isomer N(G)-nitro-D arginine methyl ester. Fos expression was also increased following the perfusion of 3-morpholino-sydnonimine, a nitric oxide donor, into the dorsal horn through a microdialysis fiber. The increased Fos was distributed within 400 microm from the edge of the microdialysis fiber. Although Fos expression was increased with 3 morpholino-sydnonimine perfusion compared to that seen with artificial cerebrospinal fluid perfusion, there was still some Fos immunostaining in the control sections. Following perfusion of artificial cerebrospinal fluid in the spinal cord of rats pretreated with N(G)-nitro-L-arginine methyl ester, it was found that Fos staining was reduced significantly compared to the control sections from animals without N(G)-nitro-L-arginine methyl ester pretreatment. These results suggest that nitric oxide helps mediate Fos expression induced by an intradermal capsaicin injection. We conclude that both endogenous and exogenous nitric oxide induce Fos expression. Involvement of nitric oxide in the development of central sensitization may affect nociceptive processing by increasing Fos expression. Since many other substances which are related to pain mechanisms can be induced by Fos, it is suggested that nitric oxide may regulate production of these substances through activation of Fos. Nitric oxide is not only involved in the development of central sensitization, but is also involved in the activation of control mechanisms affecting nociception. PMID- 10683576 TI - Anticonvulsant A(1) receptor-mediated adenosine action on neuronal networks in the brainstem-spinal cord of newborn rats. AB - Membrane potential of ventral respiratory group neurons as well as inspiratory related cranial (hypoglossal) and spinal (C(1)-Th(4)) nerve activities were analysed in brainstem-spinal cord preparations from neonatal rats. Block of Cl(-) mediated inhibition with bicuculline (plus strychnine) affected neither rhythmic depolarizations nor spike discharge in 23 of 30 ventral respiratory group cells. In the other seven neurons, block of inhibitory postsynaptic potentials evoked pronounced depolarizations and spike discharge that was synchronous with seizure like spinal nerve activity. Respiratory hypoglossal nerve activity persisted after transection at the spinomedullary junction, whereas spinal rhythm was blocked. After transection, the moderate bicuculline-evoked seizure-like perturbation of hypoglossal nerve activity was abolished and rhythmic ventral respiratory group neuron activity was not disturbed, whereas epileptiform discharge persisted in spinal nerves. The seizure-like nerve activity and depolarization of the minor subpopulation of perturbed ventral respiratory group neurons were reversed by either adenosine or the A(1) adenosine receptor agonist 2-chloro-N(6)-cyclopentyladenosine. The A(2) receptor agonist CGS 21860 had no effect. In control preparations, inspiratory nerve activity and membrane potential fluctuations (29 of 35 cells) were not changed by adenosine, 2-chloro N(6)-cyclopentyladenosine or CGS 21860. In the other six cells, adenosine evoked a hyperpolarization (<10 mV) with no major change in input resistance. The anticonvulsant effects of adenosine and 2-chloro-N(6)-cyclopentyladenosine were antagonized by the A(1) adenosine receptor blocker 8-cyclopentyl-1,3 dipropylxanthine. After pre-incubation with 8-cyclopentyl-1,3-dipropylxanthine, bicuculline also evoked seizure-like discharge in the hypoglossal nerve. The results indicate that seizure-like spinal motor output of the respiratory network upon block of Cl(-)-mediated inhibition is caused by disinhibition of spinal neuronal networks with afferent connections to the ventral respiratory group. Presynaptic A(1) adenosine receptors exert an anticonvulsant action on the disinhibited spinal motor network, but have no depressing effect per se on the isolated medullary respiratory network. PMID- 10683577 TI - The kappa opioid receptor and dynorphin co-localize in vasopressin magnocellular neurosecretory neurons in guinea-pig hypothalamus. AB - The relationship between the cloned kappa opioid receptor, dynorphin, and the neurohypophysial hormones vasopressin and oxytocin was analysed in the guinea-pig hypothalamic magnocellular neurosecretory neurons. This analysis was performed in order to understand better which population of neuroendocrine neurons in the guinea-pig is modulated by kappa opioid receptors and its endogenous ligand dynorphin. Extensive co-localization was observed between kappa opioid receptor immunoreactivity and preprodynorphin immunoreactivity in neuronal cell bodies in the paraventricular and supraoptic nuclei. Cells positive for either the kappa opioid receptor or both the kappa opioid receptor and preprodynorphin were restricted to the vasopressin expressing neuronal population and not found in the oxytocin expressing neuronal population. The kappa opioid receptor and dynorphin were examined in the posterior pituitary and both were found to be extensively distributed. Staining for the kappa opioid receptor and dynorphin B co-localized in posterior pituitary. In addition, immunogold electron microscopy confirmed that kappa opioid receptor and dynorphin B immunoreactivity were found in the same nerve terminals. Ultrastructural analysis also revealed that kappa opioid receptor immunoreactivity was associated with both nerve terminals and pituicytes. Within nerve terminals, kappa opioid receptor immunoreactivity was often associated with large secretory vesicles and rarely associated with the plasma membrane. Our data suggest that the cloned kappa opioid receptor may directly modulate the release of vasopressin but not oxytocin in guinea-pig hypothalamic magnocellular neurosecretory neurons and posterior pituitary. Furthermore, we propose that this receptor is an autoreceptor in this system because our results demonstrate a high degree of co-localization between kappa opioid receptor and dynorphin peptide immunoreactivity in magnocellular nerve terminals. PMID- 10683578 TI - Reduction of voltage-dependent magnesium block of N-methyl-D-aspartate receptor mediated current by in vivo axonal injury. AB - The post-traumatic change of the voltage-dependent Mg(2+) block of N-methyl-D aspartate response was investigated using nystatin perforated patch recording mode under the voltage-clamp condition. Motor neurons of the dorsal motor nucleus of vagus nerve were freshly dissociated from rat brain at 2h to 10 days after receiving axonal crush injuries in vivo at the neck. The reduction of voltage dependent Mg(2+) block of N-methyl-D-aspartate response became evident at more than 12h after the injury, sustained for at least five days and recovered within 10 days. Other characteristics examined such as reversal potentials, the Hill coefficient and EC(50) of N-methyl-D-aspartate-induced current were not affected by axonal injury. The Mg(2+) block of N-methyl-D-aspartate response was not affected at all by local application of colchicine onto the vagal axon in in vivo condition, suggesting that axonal injury, but not the blockade of the axonal flow, is responsible for the change of the sensitivity of N-methyl-D-aspartate response to extracellular Mg(2+). In addition, the reduction of Mg(2+) block by the nerve injury persisted regardless of the presence of protein kinase C modulators, such as 10(-6)M chelerythrine and 10(-7)M calphostin C. Therefore alteration of protein kinase C activity after axonal injury is not responsible for the maintenance of the reduced Mg(2+) block. These findings suggest that injured neurons acquire immature characteristics of plasticity with respect to the sensitivity of N-methyl-D-aspartate receptors to extracellular Mg(2+) or a long-term increase in the susceptibility to Ca(2+) excitotoxicity. PMID- 10683579 TI - Cerebral vasodilatation induced by stimulation of the pterygopalatine ganglion and greater petrosal nerve in anesthetized monkeys. AB - Although brain cell viability depends largely on cerebral circulation, mechanisms of blood flow control, such as autoregulation, or of the pathogenesis of functionally impaired blood supply to brain regions, such as in cerebral vasospasm after subarachnoid hemorrhage, have not been clearly defined. Our recent studies support the hypothesis that nitric oxide, released from nitrergic nerves, plays a crucial role as a neurotransmitter in vasodilating cerebral arteries from primate and subprimate mammals. In the present study, we demonstrated, by using arterial angiography, that electrical stimulation of the pterygopalatine ganglion produced vasodilatation of ipsilateral cerebral arteries of anesthetized Japanese monkeys. The response was abolished by intravenous injections of N(G)-nitro-L-arginine, a nitric oxide synthase inhibitor. Denervation of the ganglion elicited cerebral vasoconstriction, indicating that vasodilator nerves from the vasomotor center were tonically active. Stimulation of the greater petrosal nerve, upstream of the pterygopalatine ganglion, also elicited cerebral vasodilatation, which was abolished by treatment with the nitric oxide synthase inhibitor and with hexamethonium, indicating that the nerve is in connection via synapses with the nitrergic nerve innervating cerebral arteries. Endogenous nitric oxide released from the nerve may contribute to the maintenance of blood flow in major cerebral arteries necessary to supply blood to the different brain regions. Without this influence, cerebral arteries might be constricted to the extent that blood flow is impeded. This is the first direct evidence indicating an important role of nitric oxide liberated by pre- and postganglionic nerve stimulation in the control of cerebral arterial tone in primates. PMID- 10683580 TI - Role of the axodendritic tree in the functioning of helix bursting neurons: generation of pacemaker activity and propagation of action potentials along the axon. AB - The purpose of this study was to determine the role of the axodendritic tree in the generation of bursting pacemaker activity in the identified Helix RPa1 neuron, which is homologous to the Aplysia R15 cell, and propagation of action potentials along the axons. In doing so, I used recording of RPa1 neuron electrical activity after cutting off the right or left parietovisceral connections, the two-electrode voltage-clamp technique, registration of electrical activity of visceral nerve containing RPa1 axon branches, isolation of the RPa1 neuron and puff application of oxytocin on it. Cutting of the right (but not left) parietovisceral connection in all cases (more than 15 preparations) evoked complete disappearance of bursting pacemaker activity in the RPa1 neuron and hyperpolarization of its membrane potential up to -65 to -67 mV. Such silent state of the RPa1 neuron was maintained after its complete isolation from the ganglion. The described cutting did not result in a change of bursting activity of the pacemaker neuron V7 located in the visceral ganglion, although isolation of the V7 neuron also eliminated its own activity. Puff application of oxytocin (10 microM in a micropipette) on to the RPa1 neuron both after cutting the right parietovisceral connection or isolation of the neuron from the ganglion resulted in all cases (more than 10 cells) in transient depolarization with development of beating, oscillatory or bursting activity. Voltage clamping of RPa1 soma in the intact ganglion at a level close to zero membrane current sometimes, and, as a rule, at a more depolarized level, revealed bursting-like oscillations of membrane current, reflecting electrical bursting activity in the unclamped remote region of a neuron, most likely in the dendritic tree. Voltage clamping of RPa1 soma possessing bursting activity reveals bursting-like oscillation of membrane current and prevents propagation of corresponding axon action potentials in the visceral nerve. Controversially, clamping of RPa1 soma possessing beating activity exhibits a beating-like oscillation of membrane current and does not prevent propagation of corresponding action potentials in the nerve. Within the framework of the developed hypothesis that persistent bursting pacemaker activity of the RPa1 neuron is due to a constant activation of its peptidergic synaptic inputs [Kononenko N. I. (1993) Comp. Biochem. Physiol. 106A, 135-147], the experimental results were interpreted in the manner that these synapses and, correspondingly, the locus of electrical bursting activity generation, are localized on the dendritic tree of the RPa1 neuron mainly or possibly exclusively in the visceral ganglion. It is hypothesized that bursting and beating neuronal activities are due to functioning of different loci of the dendritic tree, regarding their electrical relations with axon branches. PMID- 10683581 TI - Acute effects of capsaicin on gastrointestinal vagal afferents. AB - Capsaicin is an important tool for investigation of thin afferent fibres, but its acute effects on subtypes of vagal afferent endings are unknown. In the gastrointestinal tract, these subtypes are: muscle endings (thought to be purely tension sensitive), mucosal endings (sensitive to stroking and chemical stimuli) and endings in the oesophagus with both properties. Acute capsaicin sensitivity was investigated in ferrets using in vivo and in vitro methods. Single-fibre activity was recorded from 63 vagal afferents: 12 Adelta-fibres, 15 C-fibres and 36 unclassified fibres with endings in the oesophagus (n=42), stomach (n=19) and duodenum (n=2). Responses to capsaicin occurred independently of motility changes and were therefore due to direct activation of the receptor ending. In the oesophagus in vivo, two of 10 tension receptors and one of one mucosal receptor responded to intraluminal application of 3.25 mM capsaicin. In the stomach and duodenum, five of 14 tension receptors and two of four mucosal receptors responded to close-systemic (32-164 nmol) capsaicin. In an in vitro gastro oesophageal preparation, three of five tension, four of 21 mucosal and two of eight tension/mucosal receptors responded to topical application of 1mM capsaicin. Occurrence of responses was therefore unrelated to location of endings and isolation of tissue. Responsiveness was also unrelated to conduction velocity. Capsaicin caused desensitization of responses to further capsaicin application in 37% of afferents. It additionally caused cross-desensitization to mechanical stimuli, which was also seen in afferents that did not respond directly to capsaicin. In conclusion, capsaicin acutely activates all subtypes of gut vagal afferents in vivo and in vitro, although responsiveness is restricted to 30% of fibres and follows no specific pattern. Acute desensitization may be induced with or without a response. PMID- 10683582 TI - Dexamethasone pre-treatment interferes with apoptotic death in glioma cells. AB - Glucocorticoids are known to influence the ability of cells to undergo apoptosis, directly inducing apoptosis in thymocytes while inhibiting it in hepatoma and carcinoma cells. Dexamethasone, a synthetic glucocorticoid, is reported to induce partial resistance to certain anticancer drugs in glioma cell lines. In the present study, the effect of dexamethasone on apoptosis of glioma and astrocytoma cell lines was investigated. Exposure of D384 human astrocytoma and C6 rat glioma cells to staurosporine induced apoptosis as judged by the formation of condensed nuclei and caspase activation. Pre-treatment of cells with dexamethasone caused a reduction in staurosporine-induced apoptosis. In addition, dexamethasone also conferred protection against the induction of apoptosis by anticancer agents including camptothecin and etoposide. The protective effect of dexamethasone was dose and time dependent, with maximal protection obtained with concentrations equal to or greater than 100 nM and a pre-incubation period of at least 24h. The earliest significant inhibition was seen with a pre-incubation period of 8h. Co treatment with the glucocorticoid receptor antagonist RU38486 abolished the effect of dexamethasone, indicating that the protection due to dexamethasone is mediated via this receptor. Dexamethasone was found to induce a time-dependent up regulation of Bcl-x(L) protein expression. However, the ability of cytochrome c/dATP to activate the caspase cascade in cytosolic extracts of D384 cells was unaffected by prior exposure of the cells to dexamethasone (1 microM) for 48 h. In conclusion, dexamethasone inhibits the induction of apoptosis in astrocytoma cells, probably via an up-regulation of Bcl-x(L), which could prevent cytochrome c release from mitochondria and subsequent caspase activation. Since glucocorticoids are often used in the treatment of gliomas to relieve cerebral oedema, the inhibition of apoptosis by these compounds could potentially interfere with the efficacy of chemotherapeutic drugs. PMID- 10683583 TI - Microglia in organotypic hippocampal slice culture and effects of hypoxia: ultrastructure and lipocortin-1 immunoreactivity. AB - Lipocortin-1 immunocytochemistry was used to study the various cell forms of microglia that appear during organotypic hippocampal tissue culture, as well as in the in vitro toxic hypoxia model. Antibodies against lipocortin-1 identified activated and phagocytic cells that were abundant in a slice after the plating of a culture: cells of the intermediate form at the later time-points of culturing, resting ramified microglia beginning from the seventh day of culturing, as well as activated and phagocytic cells that appeared in the slice after experimental toxic hypoxia induced by potassium cyanide treatment. Lipocortin-1-positive microglia cell forms corresponded well to the description of the microglia in vivo, and the morphology of microglia corresponded to the circumstances under which these cells were observed in slice cultures. Electron microscopic studies have demonstrated, for the first time, that microglia in organotypic slice culture preserve morphological features typical of different microglial forms in vivo, as well as specific contacts and interactions with the other neural tissue elements. After experimental toxic hypoxia, rapid changes in microglial ultrastructure and localization were observed, reminiscent of in vivo models of ischaemia. In conclusion, observations of microglial morphology and behaviour allow us to suggest that microglia in the organotypic culture preserve their essential characteristic features and properties, thus providing an important model system for studying the structure and function of these cells. PMID- 10683584 TI - Neurotrophin regulation of sodium and calcium channels in human neuroblastoma cells. AB - Neurotrophins, acting through tyrosine kinase family genes, are essential for neuronal differentiation. The expression of tyrosine kinase family genes is prognostic in neuroblastoma, and neurotrophins reduce proliferation and induce differentiation, indicating that neuroblastomas are regulated by neurotrophins. We tested the effects of nerve growth factor and brain-derived neurotrophic factor on Na(+) and Ca(2+) currents, using the whole-cell patch-clamp technique, in human neuroblastoma NB69 cells. Control cells exhibited a slow tetrodotoxin resistant (IC(50)=98 nM) Na(+) current and a high-voltage-activated Ca(2+) current. Exposure to nerve growth factor (50 ng/ml) and/or brain-derived neurotrophic factor (5 ng/ml) produced the expression of a fast tetrodotoxin sensitive (IC(50)=10 nM) Na(+) current after day 3, and suppressed the slow tetrodotoxin-resistant variety. The same type of high-voltage-activated Ca(2+) current was expressed in control and treated cells. The treatment increased the surface density of both Na(+) and Ca(2+) currents with time after plating, from 17 pA/pF at days 3-5 and 1-5 to 34 and 30 pA/pF after days 6-10, respectively. Therefore, both nerve growth factor and brain-derived neurotrophic factor, acting through different receptors of the tyrosine kinase family and also possibly the tumor necrosis factor receptor-II, were able to regulate differentiation and the expression of Na(+) and Ca(2+) channels, partially reproducing the modifications induced by diffusible astroglial factors. We show that neurotrophins induced differentiation to a neuronal phenotype and modified the expression of Na(+) and Ca(2+) currents, partially reproducing the effects of diffusible astroglial factors. PMID- 10683585 TI - c-Jun/AP-1 (N) directed antibodies cross-react with "apoptosis-specific protein" which marks an autophagic process during neuronal apoptosis. PMID- 10683587 TI - Gerrits M. A. F. M., wiegant V. M. And van ree J. M. (1999). Endogenous opioids implicated in the dynamics of experimental drug addiction: an in vivo autoradiographic analysis. Neuroscience 89, 1219-1227 PMID- 10683586 TI - c-Jun/AP-1 (N) expression and apoptosis. PMID- 10683589 TI - Systematic reviews of wound care management: (2). Dressings and topical agents used in the healing of chronic wounds. PMID- 10683591 TI - Factors that limit the quality, number and progress of randomised controlled trials. PMID- 10683592 TI - Antimicrobial prophylaxis in total hip replacement: a systematic review. PMID- 10683593 TI - Health promoting schools and health promotion in schools: two systematic reviews. PMID- 10683594 TI - Economic evaluation of a primary care-based education programme for patients with osteoarthritis of the knee. PMID- 10683597 TI - Obesity reduction through lifestyle modification. AB - Obesity is a worldwide public health problem. One in three Canadians is overweight, a prevalence that is already high and increasing. Moreover, 54% of men and 37% of Canadian women are characterized as abdominally obese, the phenotype that is strongly associated with cardiovascular disease and type II diabetes. These observations underscore the importance of considering the efficacy of methods commonly used to reduce total and abdominal obesity. These strategies include a decrease in energy intake (diet), an increase in energy expenditure (exercise), or pharmacological intervention. The combination of diet and exercise is more commonly prescribed, with pharmacological intervention suggested only when lifestyle changes fail to achieve weight loss. The aim of this report is to review current knowledge regarding the influence of diet and exercise as treatment strategies for obesity reduction and provide recommendations for attaining and maintaining a healthy weight. The importance of diet composition in the treatment of obesity is also considered. PMID- 10683598 TI - Exercise and training in women, Part I: Influence of gender on exercise and training responses. AB - Exercise and training responses in women are briefly reviewed. Part I of the paper considers the influence of gender on such responses. The average woman has a smaller inherent aerobic power and less muscular strength than a man, reflecting sociocultural influences, physical size, body composition, and hormonal milieu. Nevertheless, the best-trained women can out-perform sedentary men. The handicap of the average woman is offset by a lighter body mass and a tendency to metabolize fat rather than carbohydrate during exercise. A lack of anabolic hormones may limit training increases of muscle bulk in the female. A low initial fitness may enhance the scope for training tolerance, but it also limits tolerance of conditioning. Nevertheless, women seem less vulnerable than men to exercise-induced sudden death and overtraining. Part II of the review considers the influence of the menstrual cycle and pregnancy upon exercise and training responses. Physical activity programmes for young women should take account of possible pregnancy. Potential dangers to the foetus include an excessive rise of core body temperature, a decrease of maternal blood sugar, and foetal hypoxia. Nevertheless, regular moderate exercise generally has a favourable impact upon pregnancy outcomes. PMID- 10683599 TI - Exercise and training in women, Part II: Influence of menstrual cycle and pregnancy. AB - This part of the review considers the impact of the menstrual cycle and pregnancy upon exercise performance, together with the implications of continued, regular exercise for the developing foetus. Specific issues that are covered include changes in physical performance over the menstrual cycle; the impact of training on the menstrual cycle; a need for awareness of potential pregnancy; alterations in fitness, performance, circulatory, respiratory and metabolic function during pregnancy; potential hazards to the foetus from hyperthermia and hypoxia; a recommended physical activity programme for the pregnant woman; and the impact of continued exercise upon pregnancy outcomes. PMID- 10683600 TI - The efficacy of SPORT as a dietary supplement on performance and recovery in trained athletes. AB - This study investigated the efficacy of SPORT (a popular dietary supplement) in improving performance and assisting recovery in 9 trained athletes. In a double blind, crossover experiment, subjects ran at workloads of 60 and 80% of peak oxygen uptake (Peak VO2) for 5 min each with 5 min recovery after each bout and at 100% Peak VO2 until exhaustion. Two capsules of either SPORT or a gelatin placebo were administered 1 hr prior to exercise and immediately after each workload. Heart rate (HR) and blood lactate (BLa) were measured at 1 hr prior to exercise, immediately after the 100% exercise bout and at 5, 10, 20, and 45 min during recovery. No significant differences between treatments on HR and BLa measures at any of the 6 time periods, or on subjects' time to exhaustion were found. Under the conditions of this experimental design, SPORT had no beneficial effects on performance or recovery in trained athletes. PMID- 10683601 TI - The acute effects of androstenedione supplementation in healthy young males. AB - We examined the effects of androstenedione supplementation on the hormonal profile of 10 males and its interaction with resistance exercise. Baseline testosterone, luteinizing hormone, estradiol, and androstenedione concentrations were established by venous sampling at 3 hr intervals over 24 hr. Subjects ingested 200 mg of androstenedione daily for 2 days, with second and third day blood samples. Two weeks later, they ingested androstenedione or a placebo for 2 days, in a double-blind, cross-over design. On day 2, they performed heavy resistance exercise with blood sampled before, after, and 90 min post. The supplement elevated plasma androstenedione 2--3-fold and luteinizing hormone approximately 70% but did not alter testosterone concentration. Exercise elevated testosterone, with no difference between conditions. Exercise in the supplemented condition significantly elevated plasma estradiol by approximately 83% for 90 min. Androstenedione supplementation, thus, is unlikely to provide male athletes with any anabolic benefit and, with heavy resistance exercise, elevates estrogen. PMID- 10683602 TI - Immunocytochemical localization of the metabotropic glutamate receptor mGluR4a in the piriform cortex of the rat. AB - This study evaluates the localization of the metabotropic glutamate receptor mGluR4a in the piriform cortex of rats using preembedding immunocytochemical methods. At the light microscopic level, punctate labeling was evident in layers Ia and Ib of the piriform cortex, and immunolabeled fibers were present in layers II and III. Following bilateral destruction of the olfactory bulb, the density of labeled puncta in layer Ia decreased. These results suggest that the receptor is present on the terminals of the lateral olfactory tract (LOT). Electron microscopic evaluation of layers Ia and Ib revealed that mGluR4a was localized in synaptic terminals in layers Ia and Ib. The terminals had clear, round synaptic vesicles and terminated on asymmetric synapses on dendritic spines and shafts. There was also immunolabeling of some dendritic profiles in layers Ia and Ib that were postsynaptic to unlabeled presynaptic terminals. These observations suggest that mGluR4a is present on presynaptic terminals in the layers of the piriform cortex that receive LOT and associational synapses. This is the same area in which previous studies have revealed the presence of mGluR7 and mGluR8, suggesting that all three receptors may be colocalized. PMID- 10683603 TI - Expression of L1 and TAG-1 in the corticospinal, callosal, and hippocampal commissural neurons in the developing rat telencephalon as revealed by retrograde and in situ hybridization double labeling. AB - In the telencephalon, the corticospinal (CS), callosal, and hippocampal commissural neurons are the major types of neurons that have axons crossing the midline of the brain. To understand the mechanisms involved in crossing the midline structure and to examine whether the expression patterns of L1 and TAG-1 in the commissural neurons are similar to those in the spinal cord, we investigated L1 and TAG-1 expression in these neurons in rats by using a double labeling technique involving retrograde labeling and in situ hybridization. Expression of L1 messenger RNA was detected in the retrogradely labeled CS projection neurons by 1,1;-dioctadecyl-3,3, 3;,3;-tetramethylindocarbocyanine perchlorate (DiI) injection into the pons at embryonic day (E) 19, but expression of TAG-1 messenger RNA was not detected in these neurons. Also, after their axons crossed the pyramidal decussation, continued expression of L1 but no expression of TAG-1 in the CS projection neurons was shown by an additional double-labeling experiment involving DiI injection into the spinal cord at postnatal day (P) 1. An immunohistochemical study showed that L1 was continuously present in each level of the CS tract at E21 and P3, but TAG-1 immunoreactivity was not found in any level at any stage. Finally, we examined the expression of L1 and TAG-1 messenger RNAs in the callosal and hippocampal commissure neurons after their axons had crossed the midline by using the double-labeling technique. In both cases, hybridization signals of the L1 and TAG-1 messenger RNAs were observed in the retrogradely labeled neurons at P3. These results suggest that the roles of L1 and TAG-1 in the formation of the commissures in the forebrain are different from their roles in the spinal cord. PMID- 10683604 TI - Retinoid-dependent gene expression regulates early morphological events in the development of the murine retina. AB - Endogenous retinoids have been implicated in the axial patterning of the embryonic vertebrate retina; however, no studies have directly examined how asymmetric retinoid-dependent gene expression regulates early morphological events in the development of the retina. Here we used a line of indicator mice that possess a retinoid-dependent transgene to examine the relationship between retinoic acid (RA)-dependent gene expression and events occurring during early eye morphogenesis, such as the closure of the optic disc. We found that retinoid regulated gene expression shifts along the dorsal/ventral axis of the embryonic retina; at embryonic day (E) E11.5 transgene expression is restricted to the neuroepithelium in dorsal retina, and by E14.5 only immature cells located in ventral retina and the dorsal retinal margins demonstrate transgene activation. By manipulating RA levels, we were not only able to systemically alter RA dependent gene expression along the dorsal/ventral axis, but also to affect retinal morphology. In particular, reducing RA availability resulted in the abnormal closure of the optic fissure. These results indicate that asymmetric levels of RA regulate early RA-dependent gene expression in the eye and demonstrate that the normal pattern of retinoid-dependent gene transcription along the dorsal/ventral axis is critical for the proper development of the vertebrate retina. PMID- 10683605 TI - Sensory neuroanatomy of a passively ingested nematode parasite, Haemonchus contortus: amphidial neurons of the first stage larva. AB - When infective larvae of Haemonchus contortus (a highly pathogenic, economically important, gastric parasite of ruminants) are ingested by grazing hosts, they are exposed to environmental changes in the rumen, which stimulate resumption of development. Presumably, resumption is controlled by sensory neurons in sensilla known as amphids. Neuronal function can be determined by ablation of specifically recognized neurons in hatchling larvae (L1) in which neuronal cell bodies are easily visualized using differential interference microscopy. Using three dimensional reconstructions from electron micrographs of serial transverse sections, amphidial structure of the L1 is described. Each amphid of H. contortus is innervated by 12 neurons. The ciliated dendritic processes of 10 neurons lie in the amphidial channel. Three of these end in double processes, resulting in 13 sensory cilia in the channel. One process, that of the so-called finger cell, ends in a number of digitiform projections. Another specialized dendrite enters the amphidial channel, but leaves it to end within the sheath cell, a hollow, flask-shaped cell that forms the base of the amphidial channel. Although not flattened, this process is otherwise similar to the wing cells in Caenorhabditis elegans; we consider it AWC of this group. Two other neurons, ASA and ADB, appear to be homologs of wing cells AWA and AWB in C. elegans, although they end as ciliated processes in the amphidial channel, rather than as flattened endings seen in C. elegans. Each of the 12 amphidial neurons was traced to its cell body in the lateral ganglion, posterior to the worm's nerve ring. The positions of these bodies were similar to their counterparts in C. elegans; they were named accordingly. A map for identifying the amphidial cell bodies in the living L1 was prepared, so that laser microbeam ablation studies can be conducted. These will determine which neurons are involved in the infective process, as well as others important in establishing the host-parasite relationship. PMID- 10683606 TI - Glutamate-induced cobalt uptake reveals non-NMDA receptors in rat taste cells. AB - Taste receptor cells are chemical detectors in the oral cavity. Taste cells form synapses with primary afferent neurons that convey the gustatory information to the central nervous system. Taste cells may also synapse with other taste cells within the taste buds. Furthermore, taste cells may receive efferent connections. However, the neurotransmitters at these synapses have not been identified. Glutamate, a major excitatory neurotransmitter in other sensory organs, might act at synapses in taste buds. We used a cobalt staining technique to detect Ca(2+) permeable glutamate receptors in taste buds and thus establish whether there might be glutamatergic synapses in gustatory end organs. When 500 microm slices of foliate and vallate papillae were briefly exposed to 1 mM glutamate in the presence of CoCl(2), a subset of spindle-shaped taste cells accumulated Co(2+). Cobalt uptake showed concentration-dependency in the range from 10 microm to 1 mM glutamate. Interestingly, higher glutamate concentrations depressed cobalt uptake. This concentration-response relation for cobalt uptake suggests that synaptic glutamate receptors, not receptors for glutamate taste, were activated. Sensory axons and adjacent non-sensory epithelium were not affected by these procedures. Glutamate-stimulated cobalt uptake in taste cells was antagonized by the non-NMDA receptor antagonist CNQX. Depolarization with 50 mM K(+) and application of NMDA (300 microM) did not increase the number of stained taste cells. This pharmacological characterization of the cobalt uptake suggests that non-NMDA receptors are present in taste cells. These receptors might be autoreceptors at afferent synapses, postsynaptic receptors of a putative efferent system, or postsynaptic receptors at synapses with other taste cells. PMID- 10683607 TI - Focal denervation alters cellular phenotypes and survival in the developing rat olfactory bulb. AB - Several studies have demonstrated that contact between the olfactory nerve and the forebrain is critical for normal olfactory bulb development. Removal of the embryonic olfactory placode results in a failure of the olfactory bulb to form, as well as causing other forebrain malformations. The current study introduces a technique that permits removal of contact between specific regions of the olfactory nerve and the bulb early in development, without causing damage to other brain regions, and without removing the peripheral olfactory organ. The manipulation, which involves insertion of a small Teflon chip between the cribriform plate and the bulb, prohibits growth of new axons into the "shadow" region behind the implant. Focal denervation of the olfactory bulb causes a decrease in bulb and layer sizes, a reduction in mitral cell number, and changes to bulb architecture. Using a battery of antibodies (OMP, MAP2, TuJ1, calretinin, calbindin, parvalbumin, TH, and GAD), we further demonstrated that 1) focal denervation alters the relationship between the olfactory nerve and the bulb, 2) the fine structure of cells in denervated regions is disrupted, and 3) cellular phenotypes change in response to loss of afferent contact. These results suggest that contact between the olfactory nerve and the bulb is important for maintaining bulb architecture and cell survival, structure, and phenotype. They also point to focal denervation as a useful technique for examining the role of neural contact in olfactory development and maintenance of the central nervous system. PMID- 10683609 TI - Proliferation and programmed cell death of neuronal precursors in the mushroom bodies of the honeybee. AB - We have studied proliferation and programmed cell death in the brain of the honeybee during metamorphosis. DNA fragmentation detection using the TUNEL method combined with 5-bromodeoxyuridine incorporation experiments reveal that in the mushroom bodies neurogenesis is terminated by extensive apoptosis. Proliferation of mushroom body neuroblasts is active until the fourth day of pupal development, ceasing abruptly within 1 day after the onset of apoptosis in the mushroom body proliferative clusters. Inside the mushroom bodies, apoptosis spreads from the apical ends of proliferative clusters, beneath the brain's surface, toward the basal ones. The distributions of apoptotic cells and those in the S phase of the cell cycle overlap significantly. Electron microscopic analysis gives further evidence that mushroom body neuroblasts themselves undergo programmed cell death. We suggest that programmed cell death may be the main factor controlling the final number of Kenyon cells produced during metamorphosis. The overlap in time and space between proliferation and apoptosis raises the question of whether the neuronal precursors switch to programmed cell death during the progression of the cell cycle, or afterwards. PMID- 10683608 TI - Segregation of serotonin 5-HT2A and 5-HT3 receptors in inhibitory circuits of the primate cerebral cortex. AB - An emerging concept of cortical network organization is that distinct segments of the pyramidal neuron tree are controlled by functionally diverse inhibitory microcircuits. We compared the expression of two serotonin receptor subtypes, the G-protein-coupled 5-hydroxytryptamine2A receptors and the ion-channel gating 5 HT3 receptors, in cortical neuron types, which control these microcircuits. Here we show, using light and electron microscopic immunocytochemical techniques, that 5-HT2A receptors are segregated from 5-HT3 receptors in the macaque cerebral cortex. 5-HT2A receptor immunolabel was found in pyramidal cells and also in GABAergic interneurons known to specialize in the perisomatic inhibition of pyramidal cells: large and medium-size parvalbumin- and calbindin-containing interneurons. In contrast, 5-HT3 label was only present in small GABA-, substance P receptor-, and calbindin-containing neurons and in medium-size calretinin containing neurons: interneurons known to preferentially target the dendrites of pyramidal cells. This cellular segregation indicates a serotonin-receptor specific segmentation of the GABAergic inhibitory actions along the pyramidal neuron tree. PMID- 10683610 TI - Synapse distribution of olfactory interneurons in the procerebrum of the snail Helix aspersa. AB - The procerebrum is believed to be important for processing olfactory information and storing olfactory memories in terrestrial pulmonate molluscs. Previous results have demonstrated that the procerebral cell population is morphologically heterogeneous. In the present study, serial sections and electron microscopy were used to investigate differences in synapse distributions. The results demonstrate that procerebral neurons with different sites of arborization have distinct patterns of synapse distribution that probably reflect different functional contributions to the olfactory pathway. Cells that have all their arborizations in the procerebrum, but none in the internal mass, have multiple large varicosities that are specialized for output. On the other hand, cells that arborize in the internal mass or outside the procerebrum have mostly input synapses proximal to the soma and mostly output synapses in the terminal region of the neurites. These cells appear to transmit information from the procerebral cell body mass to other central nervous system regions, e.g., the internal mass and the mesocerebrum. The implications of these data are twofold. Firstly, the procerebrum directly participates in distributing processed olfactory information to more central regions of the nervous system. Secondly, the procerebral neuronal population may be divisible into two subgroups: 1) intrinsically arborizing interneurons; and 2) projection neurons. This is significant because the neural organization of the procerebrum may now be compared with that of olfactory systems in other organisms. PMID- 10683612 TI - CK-MB mass test in ischemic myocardial injury. Comparison of two tests: BioMerieux Vidas and sanofi access immunoassays. AB - The analytical and clinical performances of the new fluorescent immunoassay (CK MB mass Vidas-BioMerieux) were examined and compared to the chemiluminescent test (CK-MB mass Access-Sanofi-Pasteur). Assay precisions of the CK-MB Vidas test within-assay or between-assay were less than 5.4 and 5.3%, respectively. Linearity was tested up to 214 microg/L. The CK-MB Vidas test was free of interference with CK-BB, CK-MM, and macro-CK. One hundred nineteen blood samples from patients with ischemic myocardial injury (IMI): acute myocardial infarction (AMI), suspected myocardial contusion (SMC), and unstable angina pectoris (UA), were tested using both immunoassays. In AMI, a good correlation was found (Y [CK MB Access] = 1.1372 x [CK-MB Vidas] - 6.3902; r(2) = 0.96). In UA and SMC, low values were observed and both methods were well correlated (Y [CK-MB Access] = 1.3662 x [CK-MB Vidas] + 0.0671; r(2) = 0.97). Clinical data were in good agreement with both immunoassays. ROC analysis performed in AMI demonstrated that the clinical performances of the two assays were similar. PMID- 10683613 TI - Measurement of adenylate cyclase activity in the right ventricular endomyocardial biopsy samples from patients with chronic congestive heart failure. AB - A highly sensitive fluorometric assay technique was adopted in order to examine the adenylate cyclase activity in the minute right ventricular endomyocardial biopsy samples from patients with chronic congestive heart failure (n = 10). Norepinephrine (10(-4) M) and adenosine (10(-3) M) were incubated for 30 min with 10 microl of membrane preparation (1-2 mg protein/mg) to analyze the extent of the receptor-coupled adenylate cyclase activity. Forskolin (10(-4) M) stimulation was used to estimate the maximum adenylate cyclase activity (pmol/mg protein/min, mean +/- SE). The new microanalytical cyclic AMP assay involves four steps: enzymatic destruction of noncyclic adenine nucleotides and phosphorylated metabolites, conversion of cyclic AMP to ATP, amplification of ATP by enzymatic cycling, and fluorometric measurement of NADPH, which is generated in proportion to initial cyclic AMP levels. Basal and forskolin-stimulated maximum adenylate cyclase activities were 75 +/- 8 and 123 +/- 15, respectively. Norepinephrine increased the adenylate cyclase activity to 107 +/- 14, while adenosine tended to decrease it to 65 +/- 7. In addition, elimination of adenosine by adenosine deaminase (10 U/ml) slightly increased the adenylate cyclase activity to 82 +/- 9. These results indicate that the adenylate cyclase activity can be measured in minute endomyocardial biopsy samples. Use of this new approach shows promise of becoming a new and potentially important way to predict the efficacy of pharmacological treatment. PMID- 10683614 TI - An application of apo(a) isoforms for the clinical assessment of Lp(a). AB - To examine whether or not Lp(a) is applicable as a diagnostic marker for atherosclerosis, we studied the correlation between Lp(a) levels and molecular weights of apo(a) isoforms in sera from both normal healthy adults and diabetic patients. Serum Lp(a) level was measured by turbidimetric immunoassay (TIA) and the molecular weight of apo(a) isoform was determined by Western blotting analysis. The serum Lp(a) levels of the diabetic patients (25.0 mg/dl +/- 2.2 [mean +/- SE], n = 54) were significantly higher than those of the normal subjects (14.4 mg/dl +/- 0.57, n = 500). With respect to the correlation between serum Lp(a) levels and the molecular weights of apo(a) isoforms, there was an inverse correlation in sera from normal subjects (n = 298), whereas there was no correlation in sera from the diabetic patients. Statistical significant inverse correlation (r = -0.91, y = 224.25 - 3.07x) was especially observed in 50 representative apo(a) isotypes from the normal subjects. By applying a standardized curve based on the significant inverse correlation to serum Lp(a) levels, 40.7% (22/54) of the diabetic patients were revealed to have an abnormally high value of serum Lp(a). Moreover, it was found that the significantly higher mean value of serum Lp(a) in the diabetic group was caused by the 22 patients with higher value of Lp(a). The present findings suggest that determination of apo(a) isoform size provides estimation of the serum Lp(a) value and that the inverse correlation curve between serum Lp(a) level and the molecular weight of apo(a) isoform may be applicable to the clinical use of Lp(a). PMID- 10683615 TI - Diagnosis of quinolone-resistant Coxiella burnetii strains by PCR-RFLP. AB - A total of 12 strains of Coxiella burnetii (8 Greek isolates from acute Q-fever patients, two reference strains-Nine Mile and Q212-and two pefloxacin-resistant laboratory strains) were examined for the presence of point mutations in the quinolone resistance determining region (QRDR) of gyrA gene by direct DNA sequencing of the polymerase chain reaction (PCR)-amplified fragments. The gene sequences of all eight Greek isolates and the two reference strains Nine Mile and Q212 [minimal inhibitory concentration (MIC)A) at the corresponding codon 87 of E. coli. This mutation lead to the substitution of Glu (codon GAG) by Lys (codon AAG ). Restriction maps of amplified gyrA gene sequences were determined by GCG Wisconsin PACKAGE, and the MnlI restriction enzyme was found to cut only the sensitive strains sequences and not the resistant ones. The present PCR-RFLP analysis has proved to be a simple, rapid, and useful method for the detection of Coxiella burnetii and, at the same time, for the diagnosis of quinolone-resistant Coxiella burnetii strains. PMID- 10683616 TI - Occurrence of serum M-protein species in Japanese patients older than 50 years based on relative mobility in cellulose acetate membrane electrophoresis. AB - We investigated the occurrence of serum M-protein species in 2,007 Japanese patients older than 50 years of age. All sera samples were analyzed by cellulose acetate membrane electrophoresis. The relative mobility of an M-protein band was calculated by dividing the migration distance of M protein by that of albumin. M proteins were found to be present in 71 of 2,007 cases (3.5%). Men 80-89 years old showed the highest occurrence of M proteins, 11.0%. The relative mobility of M-protein bands, especially the band of the IgA-type M protein, increased as the patient's age advanced. The patients had higher levels of the IgG-type M protein than healthy Japanese subjects. We found that the occurrence of M-protein species in Japanese patients increases with their age. The IgG-type M protein was most frequently expressed among other types. The mobility of the M protein was greater in older patients probably because of aging-related changes in the carbohydrate chain of immunoglobulins composing an M-protein molecule. PMID- 10683617 TI - GB virus C in patients with liver disease of unknown etiology. AB - To assess the prevalence of GBV-C in patients suffering unknown liver disease we have investigated the GBV-C-RNA in serum of 54 patients: 10 with acute and 32 with chronic non-A-E hepatitis (16 active and 16 persistent), 10 with hepatocellular carcinoma, 2 diagnosed with hepatic fulminant failure, and 91 healthy blood donors (control). PCR with primers from NS3 helicase region was performed and the product was identified by a double strand DNA enzyme immunoassay. GBV appears to infect 40 and 31% of acute or chronic non-A-E hepatitis respectively. Also the GBV genome was found in 1 in 10 samples of hepatocarcinoma, in 2 cases of fulminant hepatitis, and in 1 in 91 of the control group. In spite of these results the role of GBV in the etiology of liver diseases has to be analyzed in more comprehensive studies. PMID- 10683618 TI - Immunoassays for pentamidine and related compounds: development of a facile inhibitory ELISA suitable for clinical use. AB - Aromatic dicationic drugs have a broad spectrum of activity against protozoal and fungal pathogens including Pneumocystis carinii, Leishmania mexicana amazonensis, Cryptosporidium parvum and Cryptococcus neoformans. Pentamidine serves as the exemplar for an extensive collection of newly synthesized related compounds, which have reduced toxicity and a wider range of target organisms. Assays of pentamidine and related compounds have depended on HPLC-tandem mass spectrometry (HPLC-TMS) for the quantitation and identification of drug and metabolites. Immunoassays for pentamidine would have many advantages over the HPLC methods including relative simplicity of assay format and required equipment, convenience in sample preparation and reduction in time and cost of assays. In this report we describe a simple ELISA based immunoassay for pentamidine and pentamidine-like drugs with requisite sensitivity and specificity for use as a clinical assay (EC50 value of about 50 nanomolar). Immunogen was synthesized by coupling the hapten aminopentamidine to ovalbumin (chemically modified to provide an optimal number of -SH groups) using sulfo-MBS. Maleic-anhydride activated ELISA plates were covalently sensitized using the aminopentamidine hapten and used in an inhibitory ELISA assay format whereby the ability of analyte to suppress antibody binding to sensitized plate was measured. The assay detects primarily the phenolic amidine of pentamidine when in a para position and hence can also detect structurally related derivatives of pentamidine of potential interest as new therapeutic agents. PMID- 10683619 TI - Clinical utility of a competitive ELISA to detect antibodies against Treponema pallidum. AB - Screening for Treponema pallidum infection is carried out on a large human population. To reduce costs, fewer tests which still offer adequate sensitivity and specificity could be performed. We studied the reliability of a novel indirect ELISA method to test for this infection. Several panels of sera were used that corresponded to 40 primary infections (group 1), 13 recurrences (group 2), 348 latent infections (group 3), 5 samples with anticardiolipin antibodies (group 4), 15 samples from patients with Lyme borreliosis (group 5), and 400 samples from blood donors and healthy pregnant women (group 6). The ELISA showed a global sensitivity and specificity of 100 and 99.5%, respectively. Our evaluation shows that Enzygnost Syphilis is a sensitive, specific, and simple test to screen for this infection. PMID- 10683620 TI - Psychotic disorders among inpatients with abuse of cannabis, amphetamine and opiates. Do dopaminergic stimulants facilitate psychiatric illness? AB - We have studied the occurrence of dual diagnoses (psychoses as well as abuse of either amphetamine, cannabis or opiates) during a 15-year period, among patients treated at Huddinge Hospital, Stockholm, Sweden. The purpose of the study is to evaluate if the different drugs were coupled to different rates of psychiatric co morbidity. During the period in question, 461, 425 and 371 different patients respectively had been admitted at least once due to dependency on amphetamine, cannabis and opiates. Approximately 30% of the patients with a pure abuse of amphetamine or cannabis and less than 6% of the opiate abusers had been diagnosed at least once with any of the psychoses studied. Comparing the frequency of psychoses among mixed and pure abusers of illegal drugs, with and without a concomitant abuse of alcohol, we found that the co-morbidity rate for mixed opiate abusers increased significantly from 7.2 to 20.2% when alcohol abuse was also present. For abusers of amphetamine and cannabis (both pure and mixed), no differences in co-morbidity rates were seen when an abuse of alcohol was added to that of the drugs. It is difficult to find an explanation for the significant difference between the co-morbidity of pure abuse of amphetamine or cannabis on the one hand and opiates on the other. In conclusion, our findings show that the distribution of psychotic illness is high among abusers of amphetamine and cannabis, in contrast to the generally lower co-morbidity among abusers of opiates. Although these findings are consistent with earlier studies that have shown a propensity for developing psychoses among abusers of amphetamine and cannabis, one should bear in mind that this study is based on inpatients, and is not necessarily representative for all abusers of the drugs in question. PMID- 10683621 TI - Alexithymia and anxiety: compounded relationships? A psychometric study. AB - This study is a careful examination of the relationships between different components of the alexithymia construct and state versus trait anxiety. In order to study the relations between anxiety and alexithymia in a subclinical population, we administered to 125 female college students a test battery including measures of alexithymia (TAS26), state and trait anxiety (STAI) and depression (QD2A). Results indicated positive correlations between depression, anxiety (state and trait) and alexithymia scores. Partial correlations revealed a tight link between trait anxiety and alexithymia. Furthermore, in agreement with the view that alexithymia is a multidimensional construct, the various alexithymia dimensions were found to be diversely correlated with anxiety. On the basis of partial correlation analyses, a descriptive model of the relationships between depression, state anxiety, trait anxiety and alexithymia was postulated. This model was confirmed by pathways analyses. PMID- 10683622 TI - Need for support and participation in treatment differences among subgroups of relatives to compulsorily and voluntarily admitted mentally ill individuals. AB - The need for support and participation in treatment of relatives to voluntarily and compulsorily admitted patients was addressed in a study of the quality of mental health services in two Swedish county councils. The aims of the study were to investigate differences in the above aspects between subgroups of relatives, the differences between two years of investigation, 1986 and 1991, and the differences between relatives of voluntarily and compulsorily admitted patients. The relatives investigated consisted of 79 spouses, 118 parents and 31 grown-up children. The results showed that there were only minor differences between the subgroups concerning their participation in care. Grown-up children experienced significantly less need of support and received less help for this need. In 1991, relatives participated more in the care situation, were more interested in support with regard to their own life situation, and also showed more positive attitudes towards the psychiatric services than in 1986. The relatives of the voluntarily admitted patients felt more involved in the patient's treatment, whereas the relatives of those compulsorily admitted felt less involved and perceived obstacles to admission. PMID- 10683623 TI - Psychometric qualities of the French version of the Heinrichs quality of life rating scale. AB - The quality of life concept has been increasingly used as a major tool for patient care and clinical investigations. The Heinrichs quality of life scale (QLS) is the quality of life assessment method widely used in schizophrenic patients. The QLS was translated into its French version by J.D. Guelfi according the back-translation method. This version of the validation study included 60 schizophrenic inpatients. The scale possesses acceptable psychometric qualities. The test-retest reliability is good for nearly all items of the scale and for the categories and overall score. The internal consistency alpha-coefficients were 0.9 for the global score and varied between 0.6 and 0.9 according to the category. Factor analysis elicited four factors. Convergent validity is good. Recommendations for future use of the QLS are proposed. PMID- 10683624 TI - The development and validation of a Spanish version of the quality of life in depression scale (QLDS). AB - OBJECTIVE: The adaptation of the Quality of Life in Depression Scale, the QLDS, into Spanish. METHODS: The original UK version of the QLDS was considered by two translation panels, who produced a Spanish translation. Priority was given to conceptual rather than semantic equivalence. This version was then field-tested with 15 depressed patients. The final stage of the research involved a postal survey of 62 patients, who were asked to complete the measure on two occasions. RESULTS: The Spanish QLDS was found to be appropriate and acceptable by depressed patients. The questionnaire's test-retest reliability and internal consistency were both high, and QLDS scores correlated as predicted with scores on sections of the Nottingham Health Profile. The measure was sensitive to different levels of depression as assessed by the Hospital Anxiety and Depression Scale. CONCLUSION: The Spanish version of the QLDS is suitable for use in clinical trials and for monitoring individual patients in routine clinical practice. PMID- 10683625 TI - Associations between cortical slow potentials and clinical rating scales in panic disorder: a 1.5-year follow-up study. AB - In a previous study of 15 panic patients, we demonstrated that body-related (somatic) word stimuli elicited an enhanced positive cortical slow wave compared to non-somatic word stimuli. Healthy controls did not show this difference. The present paper reports on psychometric ratings in relation to cortical slow waves in these patients. Patients were clinically reexamined after about 1.5 years. Although no significant correlations between neurophysiology and psychometric measures could be found at the onset of the study, there was a significant correlation between improvement over the follow-up period and neurophysiology. A decline in the Hamilton Anxiety Scale (HAMA), which proved to be the best estimate for improvement, was associated with the relative magnitude of the positive slow wave elicited by somatic stimuli. Our findings support cognitive models of panic disorder, which stress that abnormal processing of bodily symptoms is relevant for the development and/or maintenance of the disorder. PMID- 10683626 TI - Dysphoric subjective response to neuroleptics in schizophrenia: relationship to extrapyramidal side effects and symptomatology. AB - OBJECTIVE: Subjective reports of dysphoric responses to neuroleptic medication are common in clinical practice. However, cognitive and affective side effects of neuroleptic medications are difficult to differentiate from the symptoms of schizophrenia. We sought to elucidate the relative contribution of extrapyramidal side effects and symptomatology to dysphoric response. METHOD: Fifty clinically stable outpatients with schizophrenia attending a rehabilitation centre were assessed for extrapyramidal side effects and symptomatology before completing the drug attitude inventory (DAI). RESULTS: Presence of extrapyramidal side effects, found in 28 patients (Z = -1.99, p = 0.05), and severity of negative symptoms (r = -0.47, p = 0.001) were independently associated with dysphoric response, explaining a significant proportion of the variance (R = 0. 53, R(2) = 25.2%, F = 9.27, df = 2, p = 0.0004). CONCLUSIONS: Patients who report a dysphoric response which they associate with neuroleptic medications have more extrapyramidal side effects and more severe negative symptoms. While these responses may be part of the negative symptoms of the illness or due to other factors such as depression, we raise the possibility that they may be clinically indistinguishable from, and be a subjective measure of, the so-called 'neuroleptic-induced deficit syndrome'. PMID- 10683627 TI - Geographical analysis of the risk of psychiatric hospitalization in Hamburg from 1988 - 1994. AB - The analysis of the geographical distribution of hospital cases is obviously important for the purpose of planning hospital services, but it is of even greater significance in the planning of psychiatric services. This concern motivated our seven-year-long study, which examined hospitalization risks among various categories of psychiatric disorders in the major German city of Hamburg. Our database encompassed 77% (n = 64,000) of all psychiatric admissions in a total of 41 hospitals, most of which are general hospitals. In order to carry out the geographical analysis we employed a new statistical method based on a mixture distribution model. According to our findings, the strongest indications of an increased frequency were among male cases of schizophrenia, drug abuse and organic psychoses, and female cases of neurotic disorders, personality disorders, drug abuse and schizophrenia. We found that some areas are exposed to a risk of hospitalization for these diagnostic categories which is more than 50% above the reference. Contrary to other authors we did not identify an increased frequency of admission concentrated in the inner-city area for any of the diagnostic groups. The risk of hospitalization for schizophrenics was almost entirely associated with the close proximity of psychiatric units, while the risks for neuroses and personality disorders, as well as alcohol and drug abuse, appeared to be concentrated in areas of low social status. However, a statistically relevant correlation between an increased risk of hospitalization and low social status could be determined only for drug abuse and alcoholism. In the end, we did identify two areas in which there was an increased risk of hospitalization for several diagnostic groups, and this information will undoubtedly facilitate the planning of hospital and psychiatric services. The fact that our findings deviate to some extent from other authors - especially with respect to neuroses and personality disorders, but also to addiction - can be attributed to the inclusion of psychiatric cases from general hospitals in our geographic analysis. PMID- 10683628 TI - Comparison of the diagnosis of melancholic and atypical features according to DSM IV and somatic syndrome according to ICD-10 in patients suffering from major depression. AB - While melancholic (according to DSM) or somatic syndrome (according to ICD) has strong historical roots and substantial empirical verification, the concept of atypical features is relatively new and not sufficiently studied. The aim of the current study was to investigate the reliability of these diagnostic subcategories in patients suffering from major depression in Greece. Forty patients (eight males and 32 females) aged 19-60 years (mean 39.3, sd 12.2) suffering from major depression according to DSM-IV criteria were studied. SCAN v.2.0 was used to assess symptomatology. The presence of each criterion according to DSM-IV and ICD-10 was registered. Frequency tables were developed and factor and cluster analysis were performed. The results of the analysis suggest the existence of three syndromes which roughly reflect the melancholic and atypical but also propose a third, which can be considered as an 'undifferentiated' syndrome. The DSM demand that the existence of melancholic features be excluded first and then that diagnosis of atypical features be made was confirmed. PMID- 10683629 TI - Gender-related clinical differences in obsessive-compulsive disorder. AB - The purpose of the present study was to investigate the gender-related differences of clinical features in a sample of obsessive-compulsive (OCD) patients. One hundred and sixty outpatients with a principal diagnosis of obsessive-compulsive disorder (DSM-IV, Y-BOCS = 16) were admitted. Patients were evaluated with a semi-structured interview covering the following areas: socio demographic data, Axis I diagnoses (DSM-IV), OCD clinical features (age at onset of OC symptoms and disorder, type of onset, life events and type of course). For statistical analysis the sample was subdivided in two groups according to gender. We found an earlier age at onset of OC symptoms and disorder in males; an insidious onset and a chronic course of illness were also observed in that group of patients. Females more frequently showed an acute onset of OCD and an episodic course of illness; they also reported more frequently a stressful event in the year preceding OCD onset. A history of anxiety disorders with onset preceding OCD and hypomanic episodes occurring after OCD onset was significantly more common among males, while females showed more frequently a history of eating disorders. We found three gender-related features of OCD: males show an earlier age at onset with a lower impact of precipitant events in triggering the disorder; OCD seems to occur in a relative high proportion of males who already have phobias and/or tic disorders; and a surfeit of chronic course of the illness in males in comparison with females. PMID- 10683630 TI - Discriminant cognitive factors in responder and non-responder patients with schizophrenia. AB - To identify which improvements in cognitive function are associated with symptom resolution in schizophrenic patients treated with atypical antipsychotics. DESIGN: a prospective open trial with atypical neuroleptics (risperidone, clozapine, quetiapine). SETTING: Inpatient and outpatient units, Institute of Psychiatry. PATIENTS: Thirty-nine patients with schizophrenia according to DSM-IV criteria were included. Clinical and cognitive assessment were done at baseline (T0) and again after six months of treatment (T2). Twenty-five patients completed the trial. INTERVENTIONS: New-generation antipsychotics during six months. Patients were considered as responders if their PANSS score decreased at least 20% (n = 15) and non-responders if it did not (n = 10). OUTCOME MEASURES: a computerized cognitive assessment comprised tests of short-term-memory (digit span), explicit long-term memory (word pair learning), divided attention, selective attention and verbal fluency (orthographic and semantic). Clinical assessment included PANSS and ESRS. RESULTS: A discriminant function analysis was performed to determine which changes in cognitive performance predicted symptomatic response status. Semantic fluency and orthographic fluency were significant predictors. Together they correctly predicted responder status in 88% of cases. Memory was not a significant predictor of symptomatic response. CONCLUSION: Verbal fluency discriminated the responder from the non-responder group during a pharmacological treatment. PMID- 10683631 TI - Evidence for an increase in functional platelet 5-HT2A receptors in depressed patients using the new ligand [125I]-DOI. AB - Abnormalities in the serotonergic system have been implicated in the pathophysiology of depressive disorders. Human platelets possess serotonin-2A (5 HT(2A)) receptors, and previous research using LSD or ketanserin as ligands have indicated that their number is increased in depressed patients. Compared to other ligands previously used in platelet studies, DOI is highly selective for the 5 HT(2A) receptor and binds to its high-affinity state, therefore labeling only the receptors that are biologically coupled to the G-protein. We determined the density (Bmax) and the affinity (Kd) of 5-HT(2A) receptors labeled by [(125)I] DOI in platelets from 21 untreated patients with major depression and 21 healthy volunteers. The density of the 5-HT(2A) binding sites was found to be increased in platelets from female depressed patients as compared to controls. No changes were observed in the Kd. We did not find any relationship between the binding parameters and either the severity of the depressive episode or the suicidal tendencies of the patients. Our results show that the number of coupled platelet 5-HT(2A) receptors is increased in depressed patients, indicating that platelet 5 HT(2A) receptor function is enhanced in depression. PMID- 10683633 TI - Changing patterns of mental health care in Greece (1984-1996). AB - Greece joined the European Community in 1981 and, three years later, the Commission of the European Communities provided financial and technical assistance under EEC Regulation 815/84 for the modernisation of the traditional psychiatric care system, with the emphasis on decentralisation of mental health services and the development of community-based services, as well as on deinstutionalization of long-stay patients and improvement of conditions in public mental hospitals. Over the last 11 years, the implementation of the EEC Reg. 815/84 programme contributed to a significant shift towards extramural care and rehabilitation. The role of the large mental hospitals has gradually been diminished and a large number of long-stay patients have been deinstitutionalised. It is commonly accepted that the EEC-funded psychiatric reform programme, despite inadequacies and constraints, had an impact on the changing mental health scene in Greece. PMID- 10683632 TI - Psychotic versus nonpsychotic bipolar outpatient depression. AB - Psychotic bipolar depression was compared with nonpsychotic bipolar depression. Psychotic (n = 59) and nonpsychotic (n = 176) bipolar depressed outpatients were SCID-DSM-IV interviewed. Psychotic bipolar depression had significantly higher severity, more chronicity, fewer atypical features and axis I co-morbidity, more bipolar I, and fewer bipolar II patients. Age at onset, duration of illness, gender, and recurrences, were not significantly different. PMID- 10683634 TI - Conversion disorder: a case report of treatment with the Main Puteri, a Malay shamanastic healing ceremony. AB - We report on the case of a 38-year-old Malay housewife diagnosed with conversion disorder. It was believed that 'evil spirits' caused her symptoms. The patient was eventually treated by the Main Puteri, a Malay shamanistic healing ceremony, after previous treatments failed. The patient improved on the third day of the performance, which was attributed to the departure of the spirits from her body. This case documents the potential benefits of indigenous psychotherapy. PMID- 10683635 TI - Families: in the way? PMID- 10683636 TI - 'Why couldn't I have seen him?'. PMID- 10683637 TI - The number of partners. PMID- 10683638 TI - Setting up a home-based business. PMID- 10683639 TI - Reversing respiratory depression with naloxone. PMID- 10683640 TI - MAOIs: still here, still dangerous. PMID- 10683641 TI - Family presence during invasive procedures and resuscitation. PMID- 10683642 TI - Till death do us part. A firsthand account of family presence. PMID- 10683643 TI - Renovascular hypertension. PMID- 10683644 TI - Dialysis disequilibrium syndrome. PMID- 10683645 TI - Pharmacologic treatment of dyslipidemia. PMID- 10683646 TI - Have we forgotten the patient? PMID- 10683647 TI - How long-term care is changing. PMID- 10683648 TI - From incontinence to confidence. PMID- 10683649 TI - Optimism, pessimism, and mortality. PMID- 10683650 TI - Vaccine safety: injecting a dose of common sense. PMID- 10683651 TI - Optimists vs pessimists: survival rate among medical patients over a 30-year period. AB - OBJECTIVE: To examine explanatory style (how people explain life events) as a risk factor for early death, using scores from the Optimism-Pessimism scale of the Minnesota Multiphasic Personality Inventory (MMPI). SUBJECTS AND METHODS: A total of 839 patients completed the MMPI between 1962 and 1965 as self-referred general medical patients. Thirty years later, the vital status of each of these patients was ascertained. RESULTS: Of the 839 patients, 124 were classified as optimistic, 518 as mixed, and 197 as pessimistic. Follow-up was available for 723 patients. Among these, a 10-point T-score increase on the Optimism-Pessimism scale (e.g., more pessimistic) was associated with a 19% increase in the risk of mortality. CONCLUSION: A pessimistic explanatory style, as measured by the Optimism-Pessimism scale of the MMPI, is significantly associated with mortality. PMID- 10683652 TI - Poststreptococcal reactive arthritis in adults: a case series. AB - OBJECTIVE: To guide primary care physicians regarding the diagnosis and treatment of poststreptococcal reactive arthritis (PSReA) in adults. PATIENTS AND METHODS: We retrospectively reviewed an indexed database of all patients evaluated or hospitalized between 1976 and 1998 at Mayo Clinic Rochester and identified 35 patients with the diagnosis of reactive streptococcal arthritis, arthralgia, or arthritides. Twenty-nine patients with the diagnosis of acute rheumatic fever (ARF), septic streptococcal arthritis, or nonspecific reactive arthritis were excluded. RESULTS: PSReA was confirmed in 6 adults (3 women, 3 men; age range, 25 66 years). All patients were symptomatic with polyarthritis and oligoarthritis disproportionate to the objective findings on physical examination. Although all patients had negative throat cultures at the onset of arthritis, increased titers of anti-DNase B and antistreptolysin O confirmed recent streptococcal infection. Antecedent events included pharyngitis in 3 patients (who had received a minimum of a 10-day course of penicillin) and toxic shock syndrome in 1 patient. The latency of onset of arthritis ranged from 4 days to 6 weeks. The arthritic symptoms had a protracted course beyond the typical maximum of 3 weeks described for ARF. Treatment with aspirin did not provide symptomatic relief in any of the patients, whereas the response to therapy with nonsteroidal anti-inflammatory drugs (NSAIDs) was at least partial in all cases. Symptomatic relief occurred in 1 patient who received indomethacin and in 1 patient treated with prednisone. Penicillin prophylaxis was recommended in 1 patient. CONCLUSION: PSReA should be included in the differential diagnosis of all adult patients presenting with arthritis. Treatment strategies include aspirin, other NSAIDs, and corticosteroids. In adult patients with PSReA, there is no evidence to support the use of penicillin prophylaxis at this time. PMID- 10683653 TI - Left ventricular diastolic dysfunction in patients with hypertension and preserved systolic function. AB - OBJECTIVE: To assess prospectively diastolic function in hypertensive patients with preserved left ventricular function, particularly focusing on the limitation of the transmitral flow velocity curve alone to detect diastolic dysfunction. PATIENTS AND METHODS: Comprehensive Doppler analysis was performed in 51 hypertensive patients with preserved left ventricular systolic function. RESULTS: The ratio of the peak early diastolic filling wave velocity to the peak velocity of filling wave at atrial contraction was less than the age-adjusted mean value minus 2 SD in 16 patients, and the other 35 patients had a "normal" transmitral Doppler signal. However, the combined transmitral and pulmonary venous Doppler analysis revealed that 12 of these 35 patients had a "pseudonormal" pattern. The prevalence of diastolic dysfunction was estimated at 31% with use of transmitral Doppler alone but increased to 55% when comprehensive Doppler analysis was used (P < .05). CONCLUSION: The presence of diastolic dysfunction has been frequently overlooked in hypertensive patients with transmitral Doppler analysis alone, and an assessment of diastolic function with a comprehensive Doppler analysis is needed in patients at risk for diastolic dysfunction. PMID- 10683654 TI - Busy physicians and preventive services for adults. AB - OBJECTIVE: To study the relationship between overall productivity and the rates at which primary care physicians, in a fee-for-service setting, deliver or prescribe preventive services to adult patients. PATIENTS AND METHODS: The charts of 452 adult patients treated by 8 family practitioners and 5 internists in a fee for-service practice setting were randomly selected and abstracted for provision of 10 preventive services over a 27-month period. The percentage of eligible patients screened for each service was correlated with the production of each physician measured in relative value units (RVUs). RESULTS: The correlation coefficient between RVUs and the aggregate of the 10 services was 0.23 (95% confidence interval [CI], -0.36 to 0.70). The individual correlation coefficients between RVUs and 9 of the 10 preventive services ranged from -0.05 to 0.43. For cervical cancer screening, however, the correlation coefficient was -0.72 (95% CI, -0.91 to -0.24). CONCLUSION: With the exception of screening for cervical cancer, the data presented in this study do little to support physicians' common belief that lack of time is the reason they are unable to incorporate prevention strategies into their clinical practice. PMID- 10683655 TI - Johann Deisenhofer--Nobel Laureate in chemistry. PMID- 10683656 TI - Clinical preventive medicine in primary care: background and practice: 1. Rationale and current preventive practices. AB - Impressive evidence supports the value of clinical preventive medicine, defined as the maintenance and promotion of health and the reduction of risk factors that result in injury and disease. Primary prevention activities deter the occurrence of a disease or adverse event, e.g., smoking cessation. Secondary prevention (screening) is early detection of a disease or condition in an asymptomatic stage so treatment delays or blocks occurrence of symptoms, e.g., mammographic detection of breast cancer. Tertiary prevention attempts to not allow adverse consequences of existing clinical disease, e.g., cardiac rehabilitation to prevent the recurrence of a myocardial infarction. Preventive services have decreased morbidity and mortality from both acute and chronic conditions. However, these services are underutilized for numerous reasons. Barriers to their use include physician, patient, and health system factors. The traditional disease/treatment model should be modified to incorporate more preventive services. The subsequent 2 parts of this review will discuss suggestions for integrating primary preventive services and screening into primary care practice. PMID- 10683658 TI - Palliative care and hospice programs. AB - Palliative care and hospice programs are points on the continuum of comprehensive patient care. Unfortunately, provision of care for terminally ill patients is suboptimal. There are many new approaches to improving the skills of all physicians to fulfill the needs of patients, including better education for house staff, "train-the-trainers" programs for physicians in practice, research into methods of symptom control, and better access to established hospice programs. This review covers the history, current status, and practical suggestions for improving palliative care and hospice programs in primary care settings. PMID- 10683657 TI - Dual chamber pacing for patients with hypertrophic obstructive cardiomyopathy: a clinical perspective in 2000. AB - In some patients with hypertrophic cardiomyopathy, the dynamic left ventricular outflow tract obstructive gradient results in exercise-limiting symptoms of dyspnea, angina, and syncope. Dual chamber pacing has been proposed as a widely available alternative treatment for a subset of patients with symptomatic hypertrophic obstructive cardiomyopathy. Initial studies showed a reduction in gradient and an improvement in symptoms in almost 90% of patients with severe symptoms. We report the Mayo Clinic experience with dual chamber pacing in 38 patients with hypertrophic obstructive cardiomyopathy who had permanent pacemakers implanted for limiting symptoms intractable to medical therapy. After a mean +/- SD follow-up of 24 +/- 14 months, subjective improvement was reported in 47% of patients. However, there was no statistical difference between the maximal oxygen consumption at last follow-up and AAI pacing (atrial sensing and atrial pacing) (18.6 +/- 1.1 mL.kg-1.min-1) (i.e., when the pacemaker was implanted but not pacing continuously). This article discusses the clinical perspective on the utility of dual chamber pacing for patients with hypertrophic obstructive cardiomyopathy. PMID- 10683659 TI - Chronic obstructive pneumonia caused by a vertebral body osteophyte. AB - Osteophytes associated with spondylosis have been implicated as a cause of multiple extraspinal manifestations. Symptoms are more likely to occur with the large osteophytes associated with diffuse idiopathic skeletal hyperostosis. In the thoracic region, osteophytes have been reported infrequently as a cause of extraspinal complications. We report a case in which an anterior thoracic vertebral osteophyte was responsible for chronic obstructive pneumonia due to obstruction of the right main stem bronchus. The patient's condition improved considerably after surgical resection of the compressing thoracic osteophyte. PMID- 10683660 TI - Anemia: a cause of intolerance to thyroxine sodium. AB - Usual causes of intolerance to thyroxine sodium include coronary artery disease, advanced age, untreated adrenal insufficiency, and severe hypothyroidism. We describe 4 patients with iron deficiency anemia and primary hypothyroidism. After treatment with thyroxine sodium, these patients developed palpitations and feelings of restlessness, which necessitated discontinuation of the thyroid hormone. After the anemia was treated with ferrous sulfate for 4 to 7 weeks, they were able to tolerate thyroxine sodium therapy. Iron deficiency anemia coexisting with primary hypothyroidism results in a hyperadrenergic state. In such patients, we postulate that thyroid hormone administration causes palpitations, nervousness, and feelings of restlessness. Correction of any existing pronounced anemia in hypothyroid patients who are intolerant to thyroxine sodium therapy may result in tolerance to this agent. PMID- 10683661 TI - 59-year-old man with epistaxis, headache, and cough. PMID- 10683662 TI - The losses and suffering of terminal illness. PMID- 10683663 TI - Clinical aspects of antimicrobial resistance. AB - Soon after penicillin was introduced into clinical use, an enzyme (penicillinase) that inactivated it was discovered. Since then, the variety of antimicrobial agents has increased substantially, along with a parallel increase in resistant pathogenic microorganisms. Resistance is now recognized against all available antimicrobial agents. Factors influencing the emergence of resistance include indiscriminate use of antibiotics, prolonged hospitalizations, increasing numbers of immunocompromised patients, and medical progress resulting in increased use of invasive procedures and devices. This article provides an update on clinical aspects of a few commonly found resistant microorganisms relevant to day-to-day clinical practice. A discussion of all resistant organisms is beyond the scope of this report. Both viral and mycobacterial resistance have been addressed in previous articles in this symposium. PMID- 10683664 TI - Improvement of anemia induced by parvovirus B19 in a patient with AIDS after combined antiretroviral therapy. PMID- 10683665 TI - Dynamic left ventricular outflow tract obstruction in acute coronary syndromes. PMID- 10683666 TI - Drugs for HIV infection. PMID- 10683667 TI - VII Congress of the Bulgarian Society of Physiological Sciences. Sofia, June 10 11, 1999. Abstracts. PMID- 10683668 TI - 10th World Congress of the International Organization of Psychophysiology (IOP). Sydney, Australia, 8-13 February 2000. Abstracts. PMID- 10683669 TI - In what way does the parietal ERP old/new effect index recollection? AB - Event-related potentials (ERPs) were recorded while subjects performed a memory retrieval task requiring old/new judgements to visually presented old (previously studied) and new words. For words judged old, subjects made two binary forced choice context (hereafter source) judgements, denoting the voice (male/female) and task (action/liking) with which the test word had been associated at study. By separating the ERPs according to the accuracy of the voice and task judgements, it was possible to test the prediction that the differences between ERPs to correctly identified old and new words at parietal scalp sites (parietal old/new effects) are sensitive to the amount or quality of information that is retrieved from episodic memory (Rugg, M.D., Cox, C.J.C., Doyle, M.C., Wells, T., 1995. Event-related potentials and the recollection of low and high frequency words. Neuropsychologia 33, 471-484). In keeping with this proposal, the magnitude of the parietal old/new effects co-varied with the number of accurate source judgements. This finding is consistent with proposals that the parietal old/new effect indexes recollection in a graded fashion. PMID- 10683670 TI - Annual meeting of the Society for Adolescent Medicine. Arlington, Virginia, USA. March 22-26, 2000. Abstracts. PMID- 10683671 TI - British Association of Sports and Exercise Sciences Annual Conference. 7-10 September 1999. Abstracts. PMID- 10683672 TI - A comparative evaluation of open loop and closed loop drug administration strategies in the treatment of AIDS. AB - In recent years, many researchers in the field of biomedical sciences have made successful use of mathematical models to study, in a quantitative way, a multitude of phenomena such as those found in disease dynamics, control of physiological systems, optimization of drug therapy, economics of the preventive medicine and many other applications. The availability of good dynamic models have been providing means for simulation and design of novel control strategies in the context of biological events. This work concerns a particular model related to HIV infection dynamics which is used to allow a comparative evaluation of schemes for treatment of AIDS patients. The mathematical model adopted in this work was proposed by Nowak & Bangham, 1996 and describes the dynamics of viral concentration in terms of interaction with CD4 cells and the cytotoxic T lymphocytes, which are responsible for the defense of the organism. Two conceptually distinct techniques for drug therapy are analyzed: Open Loop Treatment, where a priori fixed dosage is prescribed and Closed Loop Treatment, where the doses are adjusted according to results obtained by laboratory analysis. Simulation results show that the Closed Loop Scheme can achieve improved quality of the treatment in terms of reduction in the viral load and quantity of administered drugs, but with the inconvenience related to the necessity of frequent and periodic laboratory analysis. PMID- 10683673 TI - Seasonal variations in water quality of an oxbow lake in response to multiple short-time pulses of flooding (Jatai Ecological Station--Mogi-Guacu River, Luiz Antonio, SP-Brazil). AB - Mogi-Guacu River is a six-order floodplain river in the upper Parana River Basin, Southern Brazil. Its yearly discharge varies from a minimum of 100 m3.s-1 to a maximum of 600 m3.s-1. Diogo Lake is a shallow lake located at its floodplain within the Jatai Ecological Station (Luiz Antonio, Sao Paulo State) and is connected throughout the year to the river through a narrow and shallow channel. The main finding of this study is that the river hidrology controls the annual variations in lake hydrochemistry through a series of hydraulic effects related to oscillations in river discharge. Lake water quality is a resultant of differential contribution from local and regional watersheds. During the low water period, lake water quality is determined by inputs from Cafundo Creek, which drains the local watershed into the lake. Raising the river level during the rain season results in the damming of lake and culminates with the entrance of river waters into the plain. The geochemistry of waters in this system is determined by weathering of sandstones with basalt intrusions. Waters are acidic (river pH = 6.00 to 7.02 and stream-lake pH = 5.15 to 6.7) and dominant cations are Na+ and K+. Major anions are almost exclusively represented by bicarbonate and an unknown concentration of organic acid anions. The overall ionic load of these soft waters in the system is therefore very low. PMID- 10683674 TI - The advancement of science in Brazil. PMID- 10683675 TI - Filling-in in topographically organized distributed networks. AB - We propose a framework for understanding visual perception based on a topographically organized, functionally distributed network. In this proposal the extraction of shape boundaries starts at retinal ganglion cells with concentric receptive fields. This information, relayed through the lateral geniculate nucleus, creates a neural representation of negative and positive boundaries in a set of topographically connected and organized visual areas. After boundary extraction, several processes involving contrast, brightness, texture and motion extraction take place in subsequent visual areas in different cortical modules. Following these steps of processing, filling-in processes at different levels, within each area, and in separate channels, propagate locally to transform boundary representations onto surfaces representations. These partial representations of the image propagate back and forth in the network, yielding a neural representation of the original image. We propose that completion takes places in a wide cortical circuit that heavily relies on V1, where long-range information helps determine contour responses at specific topographically organized locations. Neural representations of illusory contours would emerge in circuits involving primarily area V2. The neural representation of filling-in of a peripheral stimulus in a dynamic surround (such as in texture filling-in) would depend on circuits involving primarily cells in areas V2 and V3, and would include competitive mechanisms required for figure to ground segregation. Finally, we suggest that multiple representations of the stimulus engage competitive mechanisms that select the "most likely hypothesis". Such choice behavior would rely on winner-take-all mechanisms capable of constructing a single neural representation of perceived objects. PMID- 10683676 TI - Recent trends in environmental and ecological modelling. AB - The paper outlines the history of modelling and presents a status of ecological modelling: what is the modelling effort of various ecosystem and various environmental problems. Typical validation results and prognosis validation results of a eutrophication model are applied as an illustration of what models can do in environmental management. Structural dynamic modelling which considers parameters that are changed currently by optimisation of a so-called goal function is presented as one of the recent development to overcome one of the most crucial problems in modelling, namely to consider adaptation. Two case studies are presented to illustrate this approach, namely application of biomanipulation and eutrophication of a shallow lake. Forecast on the directions of development is finally presented. PMID- 10683677 TI - Identification of a new Lesch-Nyhan syndrome mutation (HPRTBrasil) and analysis of potentially heterozygous females. AB - The mutation in the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene has been determined in two brothers affected with Lesch-Nyhan syndrome. Female members of the family who are at risk for being heterozygous carriers of the HPRT mutation were also studied to determine whether they carry the mutation. DNA sequencing revealed that the boys' mother is heterozygous for the mutation in her somatic cells, but that three maternal aunts are not heterozygous. Such carrier information is important for the future pregnancy plans of at-risk females. The mutation, an A-->T transversion at cDNA base 590 (590A-->T), results in an amino acid change of glutamic acid to valine at codon 197, and has not been reported previously in a Lesch-Nyhan syndrome male. This mutation is designated HPRTBrasil. PMID- 10683678 TI - Magnetic resonance findings in amyotrophic lateral sclerosis using a spin echo magnetization transfer sequence. Preliminary report. AB - We present the magnetic resonance (MR) findings of five patients with amyotrophic lateral sclerosis (ALS) using a spin-echo sequence with an additional magnetization transfer (MT) pulse on T1-weighted images (T1 SE/MT). These findings were absent in the control group and consisted of hyperintensity of the corticospinal tract. Moreover we discuss the principles and the use of this fast but simple MR technique in the diagnosis of ALS. PMID- 10683679 TI - Possible analgesic effect of vigabatrin in animal experimental chronic neuropathic pain. AB - Since anticonvulsants have been used for treating neuralgias, an interest has arisen to experimentally test vigabatrin for its gabaergic mechanism of action. For this, 41 Wistar rats were used, and in 25 of them a constrictive sciatic neuropathy was induced (Bennet & Xie model). For testing pain symptoms, spontaneous (scratching) and evoked behaviors to noxious (46 degrees C) and non noxious (40 degrees C) thermal stimuli were quantified. Moreover, a comparative pharmacological study of vigabatrin with other analgesic anticonvulsant drugs was also performed. The results showed a possible dose-dependent analgesic effect of vigabatrin (gamma-vinyl-GABA) on experimental neuropathic pain, as shown by the significant (p < 0.05) decreasing effect of vigabatrin on scratching and by its significant (p < 0.05) increasing effect on the latency of the right hindpaw withdrawal of the animals to noxious thermal stimulus. This was corroborated by similar findings with analgesic anticonvulsants (carbamazepine, phenytoin and valproic acid). This possible and not yet described analgesic effect of vigabatrin seems not to be opioid mediated. PMID- 10683680 TI - Magnetic resonance imaging in five patients with a tumefactive demyelinating lesion in the central nervous system. AB - Five patients with a tumefactive lesion were clinically followed from 1992 to 1993. Four patients were female; age ranged from 32 to 57 years, the duration of symptoms varied from 3 days to 3 years. Neurological examination disclosed dementia in two patients, aphasia in three, hemiparesis in four, hemihypoaesthesia in three, optical neuritis in two, tetraparesis with sensitive level and neurogenic bladder in one. MRI disclosed lesions with a hypersignal on images assessed at T2 and hyposignal at T1, and gadolinium heterogeneous enhancement; these lesions were located in the: a) temporooccipital region bilaterally and brain stem, b) frontoparietal white matter, c) basal ganglia, bilateral white matter and brain stem, d) left parietal region, e) cervical spinal cord, with enlargement of this region. Cerebral biopsy was performed in three patients; acute and subacute demyelinating disease was diagnosed by histological examination. Two patients had an evolutive diagnosis; exclusion of other pathologies and clinical and radiological improvement after corticotherapy, pointed to an inflammatory disease. PMID- 10683681 TI - Multiple sclerosis in Brazil. Analysis of cerebrospinal fluid by standard methods. AB - The demonstration of intrathecal IgG synthesis has been used as an important laboratory parameter to support the diagnosis of multiple sclerosis (MS). The Committee for European Concerted Action for Multiple Sclerosis has recommended a protocol for the assessment of intrathecal IgG synthesis. We applied this methodology to determine the cerebrospinal (CSF) profile of 128 Brazilian patients with MS. We detected hypercytosis lower than 35 cells/mm3 in 97%, protein lower than 80 mg/dl in 99%, normal blood-CSF barrier function in 76%, increased IgG local production around 53% and oligoclonal IgG bands by isoelectric focusing in 85% of the definite MS patients. The diagnostic accuracy of the quantitative analysis was lower than the qualitative. The detection of oligoclonal bands was especially important in the cases of normal quantitative assays of IgG. In addition, we found a lower frequency of inflammatory reaction in CSF in our MS cases, in comparison to some European studies. PMID- 10683682 TI - Panic disorder and hyperventilation. AB - Respiratory abnormalities are associated with anxiety, particularly with panic attacks. Symptoms such as shortness of breath, "empty-head" feeling, dizziness, paresthesias and tachypnea have been described in the psychiatric and respiratory physiology related to panic disorder. Panic disorder patients exhibit both behaviorally and physiologically abnormal responses to respiratory challenges tests. OBJECTIVE: We aim to observe the induction of panic attacks by hyperventilation in a group of panic disorder patients (DSM-IV). METHOD: 13 panic disorder patients and 11 normal volunteers were randomly selected. They were drug free for a week. They were induced to hyperventilate (30 breaths/min) for 3 minutes. Anxiety scales were taken before and after the test. RESULTS: 9 (69.2%) panic disorder patients and one (9.1%) of control subjects had a panic attack after hyperventilating (p < 0.05). CONCLUSION: The panic disorder group was more sensitive to hyperventilation than normal volunteers. The induction of panic attacks by voluntary hyperventilation may be a useful and simple test for validating the diagnosis in some specific panic disorder patients. PMID- 10683683 TI - [Psychiatric symptoms among patients with dementia seen in an ambulatory service]. AB - Subjects with dementia often display an array of neuropsychiatric symptoms that include disorders of mood, delusions, hallucinations, vegetative symptoms and psychomotor abnormalities. The present study was designed to investigate the prevalence of psychiatric morbidity amongst patients with the clinical diagnosis of dementia (ICD-10) assessed at a Memory Clinic in Sao Paulo-Brazil between February 1997 and May 1998. The mental and cognitive state of patients were assessed with an extended version of the SRQ-20 and the MMSE respectively. Thirty four (45.3%) out of a total of 75 subjects scored 8 or more on the SRQ-20, indicating the presence of significant psychiatric morbidity. Depressive symptoms were reported by 69.3% of patients. Persecutory ideas and auditory hallucinations were observed in 20.0% and 16.0% of the sample respectively. Eight subjects (10.7%) described suicidal ideation--they all displayed depressive symptoms. Patients with scores on the SRQ-20 > or = 8 or who described suicidal ideation were significantly younger than their counterparts. Auditory hallucinations were more frequent amongst subjects with lower MMSE scores. There were no sex differences in the distribution of the psychiatric symptoms under investigation. The assessment of patients with dementia should always include a detailed psychiatric examination, as the detection and treatment of such symptoms may contribute to decrease the stress of patients and the burden on carers. PMID- 10683684 TI - [Facial recognition and autism]. AB - Through the presentation of four facial expressions' illustrations, we evaluate the capacity of autistic children recognition, comparing with normal intelligence children and adults. The comparison of results was accomplished through the qui square test. The differences observed were significant, showing that a disturbance of the facial expressions' perception is present in autistic children, and that it interferes directly in the social relationships. PMID- 10683685 TI - [Brain hypoxic-injury of hemorrhagic pattern in newborns: analysis of 1028 autopsies from 1960 to 1995]. AB - Hypoxic-ischaemic injury of the central nervous system (CNS) in newborns is a very prevalent entity affecting 1 to 6 children per 1000 births. This injury may induce severe neurological sequelae. We present the analysis of 1028 consecutive cases of hypoxic-ischaemic CNS injuries of haemorrhagic pattern detected in autopsies performed at the Division of Anatomic Pathology, Hospital de Clinicas, University of Parana, Brazil, from 1960 to 1995. The prevalence of these lesions was high (49.73%) amongst all autopsied newborns. The main types of haemorrhage were microscopical intra-parenchymal haemorrhages, intraventricular and periventricular haemorrhages and subarachnoid foci of bleeding. Our results emphasize that premature children constitute a high risk group for CNS haemorrhage needing special preventive therapeutic procedures to avoid neurological complications. PMID- 10683686 TI - [DNA ploidy in astrocytoma. Study in 66 Brazilian patients]. AB - The determination of the nuclear DNA content (S-phase fraction (SPF) and ploidy) was carried out by image analysis (IA). The morphometric and densitometric features of Feulgen-stained nuclei were determined from stored formalin-fixed and paraffin-embedded specimens of 66 patients with intracranial astrocytomas. Our results suggest a strong relationship between patient age, histological grade, survival and DNA ploidy and SPF. The analysis of the proliferative activity of intracranial astrocytomas is very helpful in understanding biological stractification and prognostication to assess tumor behaviour and planning of treatment strategies. PMID- 10683687 TI - [Congestive brain swelling in victims of fatal road accident. Frequency and association with other head injury lesions]. AB - A morphological study, macro and microscopical, was made of brain lesions in 120 victims of fatal road traffic accidents. Congestive brain swelling occurred in 21 (17.5%) patients. Owing to the brain swelling that increases the brain volume, an increase of brain weight was also observed. Brain contusion was the most frequent lesion associated with congestive brain swelling (76.2%), while the intracranial haematomas were observed in almost half of the cases. PMID- 10683688 TI - [Burst lobe in victims of fatal road traffic accident. Frequency and association with other head injury]. AB - A morphological study, macro and microscopical, was made of brain lesions in 120 victims of fatal road traffic accidents. Burst lobes were identified in 12 (10.0%) of the patients. It occurred in the frontal lobe in 6 (50.0%) patients, in the temporal lobe in 2 (16.7%) and in both lobes in 4 (33.3%) patients. A skull fracture occurred in 8 (66.7%) patients and intracranial hypertension occurred in half of cases. Nine patients were admitted in coma and three patients died immediately after the road traffic accident. All cases of burst lobes were associated with diffuse axonal injury, which explains the severe alteration of consciousness observed at the patients' admission. PMID- 10683689 TI - [The involvement of the brachial plexus in cardiac surgery with median sternotomy for the revascularization of the myocardium: clinical evaluation]. AB - To evaluate the involvement of brachial plexus in cardiac surgery with median sternotomy for the revascularization of the myocardium 113 patients (87 men and 26 women) were clinically examined in the preoperative and between the fifth and eight post-operative days. The internal thoracic artery was used in 65 of the 113 patients. The electroneuromyography was not effected in any of the patients. A lesion of the brachial plexus was found in three patients though the internal thoracic artery was used in only one patient. We believe that factors such as posture of the patient, hypothermia, thoracic braces and use of the internal thoracic artery are relevant in the lesions. Hence one must be attentive to all the factors mentioned above so as to avoid or minimize the lesions. PMID- 10683690 TI - [Medulloblastoma in adults: analysis of a casuistics and surgical results]. AB - We report on 15 cases of medulloblastoma of adult onset (8 male and 7 female) operated upon posterior fossa approach from February 1988 to October 1995. Tumors were localized in cerebellar hemisphere in 7 cases (one with extension to supratentorial notch and another case reaching the cerebello-pontine angle cistern), in vermis and hemisphere in four, only in vermis in another four. Resection was total in seven patients, subtotal in other seven, and partial in one. There was no operative mortality. Aspects regarding biological behavior, diagnosis, pathological findings, surgery and survival are discussed as well as prognostic factors. PMID- 10683691 TI - [Clinical features of transformed migraine]. AB - Most daily headache patients seen in specialized clinics present a past history of migraine. Some authors refer to it as transformed migraine and emphasize its milder intensity and clinical characteristics different from migraine. The aim of this study was to evaluate the clinical presentation of the daily headache in patients with prior history of migraine. We studied retrospectively 215 patients. We observed that a significant percentage of the patients presenting the so called transformed migraine, reported frontal and/or temporal bilateral pain and had pressure or tightening pain, which is a characteristic of chronic tension type headache. It emphasizes the loss or changing of the standard migraine features. The pulsatile pain quality remained as an important feature, specially for those with intermittent typical migraine attacks. PMID- 10683692 TI - [Management of arterial hypertension in patients with acute ischemic stroke]. AB - PURPOSE: We aimed with study to assess the current clinical practice about the management of high blood pressure in patients in the acute phase of ischemic stroke. We also comment some topics of ischemic stroke treatment. METHODS: A case report of a patient admitted 8 hours after onset of ischemic stroke and with blood pressure of 186 x 110 mmHg was presented to 120 surgeons and clinician. They were asked to decide the best therapeutic option: to increase, decrease or maintenance blood pressure. RESULTS: Thirty-eight physicians (31.7%) considered decreasing blood pressure the best therapeutics, 82 (68.3%) considered maintenance and none decided to increase it (p < 0.05). There was no difference between the two specialties conduct. The physicians, with more than 10 years of graduation, had a tendency to decrease the blood pressure (p < 0.05). CONCLUSION: The maintenance of blood pressure may present a sufficient blood support to compensate brain flow. A high percentage of the physicians (31.7%) do not know about the current concepts of therapeutics considering hypertension in acute ischemic stroke. The development on special units to treat these patients ("stroke units") may eventually decrease the morbimortality rates of ischemic stroke. PMID- 10683694 TI - Cerebrovascular disease in newborn infants. Report of three cases with clinical follow-up and brain SPECT imaging. AB - The clinical and neurological findings of three neonates with the diagnosis of cerebrovascular disease are reported. The neuropsychological evaluation disclosed impairment of fine motor function, coordination, language, perception and behavioral disturbances. Brain SPECT imaging revealed perfusional deficits in the three cases. PMID- 10683693 TI - [Infantile neuroaxonal dystrophy: report of 2 cases]. AB - We describe two cases of infantile neuroaxonal dystrophy, which is a rare, neurodegenerative disease, with autosomal recessive inheritance. The first case was an 8 year old boy, with arrested motor and mental development, ataxia and muscle weakness. On physical examination there was horizontal and vertical nystagmus, optic disc atrophy, hypotonia; deep tendon reflexes were absent. The second case was a 1.6 year old boy with arrested motor and mental development, and seizures. On physical examination there was optic atrophy, hypertonia and hyperreflexia. Both patients had on sural nerve biopsy neuronal enlargement, consistent with neuroaxonal dystrophy. Diagnosis without pathological confirmation with neuroaxonal spheroids is very difficult, because the clinical picture is variable and the neurophysiological findings are non specific. PMID- 10683695 TI - [Transitory cerebellar mutism: report of 2 cases]. AB - We present two cases of mutism observed after resection of tumors of the cerebellum, in two children of the feminine sex, being in the first case of medulloblastoma and in the second of juvenile astrocytoma. In both patients there was pre-operative lesion of low cranial nerves. The pathophysiology of the mutism involves anatomical, vascular and emotional factors, being its essential characteristics discussed with base in revision of the literature. PMID- 10683696 TI - Kearns-Sayre syndrome "plus". Classical clinical findings and dystonia. AB - We present a boy of eight years of age with symptoms of Kearns-Sayre syndrome (KSS) characterised by ophthalmoparesis, palpebral ptosis, mitochondrial myopathy, pigmentous retinitis, associated to short stature, cerebellar signs, cardiac blockade, diabetes mellitus, elevated cerebrospinal fluid protein concentration, and focal hand and foot dystonia. The skeletal muscle biopsy demonstrated ragged red fibers, cytochrome C oxidase-negative and succinate dehydrogenase-positive fibers. The magnetic resonance imaging showed symmetrical signal alteration in tegmentum of brain stem, pallidum and thalamus. Mitochondrial DNA analysis from skeletal muscle showed a deletion in heteroplasmic condition. The association of dystonia to KSS, confirmed by molecular analysis, is first described in this case, and the importance of oxidative phosphorylation defects in the physiopathogenesis of this type of movement disorder is stressed. PMID- 10683697 TI - Malaria and stroke. Case report. AB - Malaria is a parasitic disease with high prevalence in several regions of the world. Infestation by Plasmodium faciparum can, in some cases, affect the central nervous system producing encephalitis resulting in death or neurological sequelae. The mechanisms involved in the pathophysiology of the cerebral lesion are not totally clear and there are currently two theories (mechanical and humoral) concerning this. We report a case of malaria with an atypical evolution, with a stroke lesion in the territory of the middle cerebral artery, with no association with encephalitis. We conclude that the mechanical theory is the one applicable to this patient. PMID- 10683698 TI - [Thrombosis of the internal artery secondary to soft palate injury: case report]. AB - Stroke following intraoral trauma is a rare complication of a common childhood injury. In the literature these complications have been well documented, however this condition is still infrequent. In order to alert the physicians about this possible injury we report our experience with one case. Computer tomography and magnetic resonance imaging evidenced complete occlusion of the internal carotid artery. Pathogenesis of this oral trauma is discussed. PMID- 10683699 TI - [Probable Rasmussen's syndrome. Case report]. AB - The syndrome of chronic encephalitis with epilepsy (Rasmussen's syndrome) typically occurs in children and is characterized by the development of intractable focal seizures, progressive hemiparesis and intellectual deterioration. The etiology is unknown, and the pathological abnormalities vary. We report the case of a 17-year-old boy, presenting with clinical features typical of probable Rasmussen's syndrome. We discuss the etiology and treatment for this pathology. PMID- 10683700 TI - [Comparison of the neuropsychological assessment in a girl with bilateral cerebrovascular disease (moyamoya) before and after surgical intervention]. AB - Moyamoya is a chronic progressive cerebrovascular disease with characteristic angiographic findings and a clinical picture with episodes of transient ischemic attacks, headache, seizures, hemiparesis, which may resolve after surgical treatment. We describe the case of a girl with the typical findings of the disease, comparing them before and after surgery with the use of neuropsychological tests, neurological examination and laboratory tests. PMID- 10683701 TI - [Autogenic bone plug to seal burr holes: technical note]. AB - Many neurosurgical procedures can be performed by a single burr hole: neuro endoscopy, microvascular decompression, stereotactic procedures, chronic subdural haematomas. It is technically difficult to suture and close the dura, located at the bottom of such holes, which can lately lead to CSF leakage. On the other hand, the surgical material used to seal the burr holes can be divided in heterogenic (metal screws, silicon plugs, gelfoam, bone wax, metilmetacrilate, hydroxyapatite), and autogenic (fat, aponeurosis, muscle, and bone dust from trephination). The heterogenic group always brings the possibility of foreign body reaction, which can complicate the procedure lately, ensuing a new surgical procedure to clean up the area. It also favors infection. We present a simple, economic, and biologically compatible "autogenic bone plug" to seal burr holes using the bone dust from the original trephination packed with surgical. The incidence of CSF fistulae on that procedures performed by a single burr hole lowered to almost zero, and foreign body reaction was not observed to present. PMID- 10683702 TI - Cerebral systems in the pathogenesis of endogenous psychoses. 1974. AB - Mental process imply a harmonious functioning of psychic systems, assembled into larger units, psychic spheres (Table 1). Their neurophysiological representatives are brain systems of areas and pathways (Figs 1-4). Under functional and/or organic disturbances these systems originate the leading mental symptoms (Table 2) characterizing the diverse endogenous psychoses: hence, the latter's distinctive patterns. Accordingly, understanding and classification of psychoses should rest on the pathogenic dynamisms, not on clinical description. This is why Kleist's and Leonhard's conceptions of the endogenous psychoses surpass any other to exist. Kleist stands among the founders of psychiatry, by describing the "degeneration psychoses" and many single psychoses, as well as redefining, isolating and clarifying the progressive ones, later on renamed as schizophrenias (Table 3). Such pathogenic criterion may also be useful to define mental conditions other than psychoses, as hysteria, neuroses and psychopathic inferiority (Tables 4 and 5). One should consider here, besides the psychic systems and spheres involved, the way they were caught and the corresponding developmental phase. In Kleist's "degeneration psychoses"--cyclic or episodic (Table 6) the systems and spheres are disturbed by functional transient processes due to latent dispositions, while his and Leonhard's schizophrenias (Table 7) show a rather progressive, deteriorating course. The nature of the disorder is itself genetically determined, as is either its confinement to one sphere or its spreading out. The spread out pattern, while exceptional in schizophrenia, represents a rule for the "degeneration psychoses", in discussant's mind. Both groups may have symptoms alike by involvement of the same sphere (Table 8), but proper diagnosis is reached by taking pathogenesis into consideration. PMID- 10683703 TI - [Homage--Anibal Silveira (1902-1979)]. PMID- 10683704 TI - Reactions to prenatal testing: reflection of religiosity and attitudes toward abortion and people with disabilities. AB - To better understand factors associated with prenatal testing decisions, we asked individuals what they would do if through prenatal testing they discovered that they (or their partner) were carrying an affected fetus. Respondents were more uncertain about whether to continue the pregnancy when the fetus was diagnosed as having Down syndrome than when the fetus had spina bifida or hemophilia and less certain about continuing a fetus with spina bifida than one with hemophilia. There was modest support for the hypothesis that negative attitudes toward people with disabilities would be associated with an increased likelihood of choosing abortion. Religious affiliation was associated only with the decision concerning the fetus with hemophilia; church attendance was associated with the decisions concerning fetuses with all three diagnoses. PMID- 10683705 TI - Effects of modified orthography on the identification of printed words. AB - Research has shown that instructional methods involving pairing pictures with print interfere with identification of written words. Preliminary evidence, however, indicates that use of modified orthography (where a line drawing is superimposed upon the printed word) may be effective for reading instruction with individuals who have mental retardation. In the present study, we used a single subject parallel treatments design with 4 adults who had moderate to severe mental retardation. They received reading instruction under two conditions- traditional and modified orthography. Results showed that traditional orthography was a more effective method for word identification. The relative advantage of modified orthography over traditional orthography for individuals with mental retardation was not supported. PMID- 10683706 TI - Problem behaviors associated with 15q- Angelman syndrome. AB - Caregivers of persons with Angelman syndrome completed the Aberrant Behavior Checklist and Reiss Screen for Maladaptive Behavior. Seventy-three replies were received, and comparisons were made with other published data. Responses indicated that 15q- Angelman syndrome is associated with such problems as lack of speech, overactivity, restlessness, and eating and sleep problems. Episodes of inappropriate laughter were only reported for 57%, despite being considered a cardinal feature of the syndrome; eating problems (64%) and a fascination with water (68%) were reported more frequently. Overactivity was more of a problem for children; Aberrant Behavior Checklist Factor IV (Hyperactivity) was negatively correlated with age. Scores were mostly lower than for previously studied etiological groups. Therapeutic effort should be put into programs to address these problems. PMID- 10683707 TI - Models of child-family interactions for children with developmental delays: child driven or transactional? AB - Child-driven and transactional models of child-family interactions were tested with 80 children who had developmental delays and their families. Children's cognitive competence, personal-social competence, behavior and communication "hassle," and family accommodations to the children were assessed at child ages 3, 7, and 11. Accommodations were summarized as internal (within the family) and external (use of outside resources) intensity and types. Results indicate that the longitudinal relationships between children's cognitive competence, personal social competence, behavior and communication hassle, and family accommodations are best explained by a child-driven model. Implications for early intervention and for the need to consider both child and family outcomes are discussed. PMID- 10683708 TI - Influence of feature training on the formation of exemplar-specific representations in children with mental retardation. AB - Whether exemplar-specific or prototype representations were primarily used by children with mild mental retardation in a categorization task consisting of two groups of make-believe animals whose features were ill-defined was examined. Categorization performance of familiar, prototypic, and novel test phase exemplars indicated that these children were able to acquire and use both types of representations. A feature-labeling training procedure, expected to promote the formation of exemplar-specific representations prior to exemplar classification, was also incorporated within the current study's design. It was unclear whether such training had any impact on the formation of exemplar specific representations. PMID- 10683709 TI - Behavioural effects of fluoxetine and tianeptine, two antidepressants with opposite action mechanisms, in rats. AB - The behavioural effects of two antidepressants with opposite molecular mechanisms, tianeptine 7-[(3-chloro-6,11-dihydro-6 methyldibenzo[c,f][1,2]thiazepin - 11-yl)amino]heptanoic acid S,S-dioxide, CAS 66981-73-5) 5 mg/kg p.o., a serotonin reuptake enhancer, and fluoxetine (+/-)-N methyl-3-phenyl-3-[(alpha, alpha, alpha-trifluoro-p- tolyl)oxy]propylamine, CAS 54910-89-3) 5 mg/kg p.o., a serotonin reuptake inhibitor, were compared after single and prolonged administration (7 and 14 days) once daily). In all experiments the drug effects were noted at the peak activity time: 30 min after tianeptine and 60 min after fluoxetine administration. In the immobility time test both drugs had a shortening effect on immobility time only after prolonged administration or, in single treatment, after joint administration. A different pattern was observed in the two compartment test: both antidepressants showed anxiolytic effects after single and prolonged treatment. However, when the drugs were given in joint administration, the anxiolytic effects were entirely abolished after single as well as prolonged treatment. In reference spatial memory test (food finding time in the maze) tianeptine had no effect, whereas fluoxetine caused, after single and prolonged treatment, a very marked improvement of reference memory. Joint administration of both drugs resulted in worsening the effects on memory in comparison to fluoxetine alone, but the results were still significantly better vs. control. In the test for sedative action (in the Activity Meter AM-1, where the movements of the animals are counted electronically) only after prolonged treatment with tianeptine a diminished locomotor activity could be observed. It is concluded that in the action of the drugs (beside the effect on serotonin uptake) other mechanisms must play an important role. The diminished locomotor activity after tianeptine suggests an influence on the dopaminergic or GABA-Receptor system. PMID- 10683710 TI - Muscarinic properties of compounds related to arecaidine propargyl ester. AB - A series of new analogues of the arecaidine propargyl ester (CAS 35516-99-5), APE, 1a) with alcohols consisting of 4 or 5 carbon atoms were investigated at muscarinic receptor subtypes. The muscarinic activity of the quaternary and tertiary salts of the APE-related compounds were assayed on the isolated guinea pig ileum (M3 receptor subtype) and guinea-pig left atria (M2 receptor subtype) as well as on rabbit isolated vas deferens (M1 receptor subtype). The structural variations made in the APE molecule, replacing the triple bond in the ester side chain with structures such as double bond, an allene moiety, a single bond, a cyclopropyl group or two triple bonds should alter the selectivity and potency in favour of the M2 subtype. Enhanced, though modest, selectivity for M2 receptors was achieved with the 2-butynyl ester 2a. The other structural variations resulted in a loss of potency, but not necessarily of efficacy. PMID- 10683711 TI - Neurosedative and antioxidant activities of phenylpropanoids from ballota nigra. AB - Ballota nigra is a European plant known for its neurosedative properties. In this study, the ability of five phenylpropanoids (verbascoside, forsythoside B, arenarioside, ballotetroside, and caffeoyl malic acid) isolated from a hydroalcoholic extract, to bind to benzodiazepine, dopaminergic, and morphinic receptors was investigated. To carry out these studies, affinity tests with rat striata, entire brains and receptor rich preparations were employed. In addition, the phenolic aspect of these five phenylpropanoid esters led to investigate antioxidant activities using cell-free experiments and cellular experiments including isolated polymorphonuclear neutrophils (PMN). Effects of phenylpropanoid esters against reactive oxygen species as superoxide anion, peroxide hydrogen, hypochlorous acid and hydroxyl radical were tested. These molecules are liberated by PMN during inflammatory disorders, so that reproduction of this process in vitro stimulating PMN by chemical stimulants was undertaken. Results show that four of the five compounds are able to bind to the studied receptors. Inhibitory concentrations at 50% were determined and vary from 0.4 to 4.7 mg/ml. This may be in relation with the Ballota nigra known neurosedative activities. Results concerning antioxidant investigations evidence an ability to scavenge reactive oxygen species. Inhibitory concentrations at 50% obtained are comparable to those of known antioxidant drugs (mesna or N-acetyl cysteine). Moreover, the use of different stimuli having various pathways of action on PMN oxidative metabolism permits to establish that each phenylpropanoid ester has its own particular way of action by using proteine kinase C or phospholipase C pathways. PMID- 10683712 TI - Effects of tramadol and tilidine/naloxone on oral-caecal transit and pupillary light reflex. AB - As has been demonstrated in binding studies the two opioids tilidine (CAS 27107 79-7)/naloxone (CAS 357-08-4) and tramadol (CAS 36282-47-0) differ in regard to their affinities to the opioid receptor site. Therefore it is of interest to evaluate whether such a difference in opioid affinity is also seen in the pharmacological effects of clinically relevant doses in man. Following institutional approval by the local ethical committee and informed consent, 12 volunteers received oral doses of tramadol (100 mg), tilidine/naloxone (100 mg) and placebo, respectively, in a randomized, double-blind cross-over design. In order to determine the degree of constipation, oral-caecal transit time was measured using the H2-exhalation test. Additionally, in order to evaluate a centrally mediated effect, the response of the pupil to light was quantified using the pupillary light reflex technique. Both, peripheral and central mediated effects were compared to placebo. Tramadol as well as tilidine/naloxone induced a significant (p < 0.05) prolongation of oral-caecal transit when compared to placebo. However, prolongation of oral-caecal transit was significantly longer in the tilidine/naloxone (p < 0.05) than in the tramadol group. Compared to tramadol, the pronounced constipating effect of tilidine/naloxone is likely to be due to the 10 fold higher affinity of that drug to the peripheral opioid receptor sites in the intestinal tract, which are responsible for normal propulsion. Such difference in binding is underlined by a central effect, the pupillary light reflex response. The amount of constriction of the iris to light was reduced after both opioids. Again, tilidine/naloxone significantly reduced (p < 0.001) the pupillary light reflex when compared to tramadol. Other side effects such as tiredness, nausea, emesis and dry mouth were more often reported after tilidine/naloxone than after tramadol (40% versus 15%; p < 0.05). Vertigo and perspiration were more often reported after tramadol than after after tilidine/naloxone (58% and 78% versus 8%; p < 0.01). All these data support the findings that while tramadol is considered an opioid, it does not mediate its main clinical relevant properties via binding at the opioid receptor. More likely, due to its monoaminergic reuptake mechanism, to a lesser extent opioid like effects are induced. PMID- 10683713 TI - Preliminary acute and subchronic toxicity studies of GLG-V-13, a novel class III antiarrhythmic agent, in mice. AB - The acute and subchronic toxic effects of GLG-V-13 (3-[4-(1H-imidazol-1 yl)benzoyl]-7-isopropyl-3,7-diazabicyclo[3.3.1]nona ne dihydroperchlorate, CAS 155029-33-7), a novel class III with some class Ib antiarrhythmic activity, were investigated in mice. The estimated LD50 for GLG-V-13 given orally were 419 mg/kg for male mice and 383 mg/kg for female mice, respectively. The acute toxic signs appeared to be of the central nervous system in origin. Four groups of mice (15 per sex, group and dose) were fed daily with diets containing GLG-V-13 for 90 consecutive days. The equivalent daily doses were 0, 22, 50 and 121 mg/kg/day and 0, 27, 60 and 136 mg/kg/day for male and female mice, respectively. All of the mice survived. Food consumption was decreased. However, mean body weight and body weight gain were not significantly changed. Gross pathological changes, especially in the lungs and liver, were found in the middle and high dose groups. Consistent increased mean corpuscular hemoglobin concentration and decreased mean corpuscular hemoglobin were observed in all dose groups. Hepatocellular necrosis was found in both male and female mice treated with the drug and was dose dependent. Marked vacuolation of the X zone in the adrenal gland with mild to moderate deposition of ceroid pigments (brown degeneration) was observed in female mice. Lesions in the kidneys and adrenal glands may be a possible reason for changes in serum sodium and potassium ions concentrations leading to an increase in water intake. A significant reduction in cholesterol in the high dose group may be a favorable pharmacological effect of GLG-V-13. The data from the 90 day subchronic toxicity studies indicate that GLG-V-13 appears to have limited systemic toxicity potential. PMID- 10683714 TI - [The effect of a nasal spray consisting of a standardized mixture of citrus limon (succus) and an aqueous extract of Cydonia oblongata (fructus) on nasal mucociliary clearance]. AB - In a three-way-crossover study in 18 healthy male and female subjects aged from 20 to 49 years the influence of a 1% and 3% solution of a standardized composition of Citrus limon, succus, and extract from Cydonia oblonga, fructus (Gencydo) on the intranasal mucociliar clearance was investigated after multiple administration. The pH of the solution was about pH 2.3-3.2. The dose regimen consisted of 20 puffs (0.13 ml per puff) in each nostril within 24 h, which was by factor 3-10 higher than the usual therapeutic dosage of 2-6 puffs per nostril and 24 h. The mucociliar transport time was measured by a modified saccharin test, where 1 microliter of a 3-molar aqueous sodium saccharinate solution was applicated at the inferior nasal turbinate 1 cm from its anterior end using a glass capillary microliter pipet. This test was performed at screening examination, before each administration period, directly after each 24 h treatment period and 24 h after the end of each treatment. The time of initial taste perception could be defined with high precision by the volunteer since it appeared very spontaneously. Neither after intranasal administration of the 1% and 3% Citrus/Cydonia solution nor after placebo solution a prolongation of the perception time was found. It could be concluded that there is no measurable influence of the test products on the intranasal ciliar function. PMID- 10683715 TI - Pharmacokinetics of diclofenac after oral administration of its potassium salt in sachet and tablet formulations. AB - This paper reports the results of a pharmacokinetic study involving 24 healthy volunteers and designed to characterise the rate and extent of diclofenac absorption after the administration of a single dose of diclofenac (CAS 15307-86 5) potassium salt 50 mg in sachet (Voltfast) and tablet (Cataflam) formulations. Timed plasma concentrations of diclofenac during a 12-h-period after dosing were measured by means of HPLC with UV detection at 275 nm and a quantification limit of 10 ng/ml; the method was fully validated for pharmacokinetic purposes. These plasma concentrations were used to calculate Cmax, tmax, trapezoidal AUC0-t and AUC0-infinity and t1/2 by means of noncompartmental analysis. Cmax and tmax are the parameters expressing the rate of absorption, whereas the AUCs reflect the extent of absorption. The rate of absorption with the sachets proved to be very fast, reaching peak values at 10 min in seven subjects and at 15 min in the remaining subjects: mean time was 13.68 min, with concentrations at 5 min being 38% of Cmax. The average time to peak concentration with the tablets was 53.10 min. The extent of absorption of the sachets and tablets was similar, with AUC0 infinity values of respectively 1362 and 1214 ng.ml-1.h, and a 90% confidence interval 1.05-1.20. The highly soluble potassium salt of diclofenac was rapidly absorbed, especially in its sachet formulation, and thus appears to be an invaluable analgesic agent that is particularly useful for quick pain relief. PMID- 10683716 TI - Synthesis and biological evaluation of indole containing derivatives of thiosemicarbazide and their cyclic 1,2,4-triazole and 1,3,4-thiadiazole analogs. AB - New indolic derivatives of thiosemicarbazides and some cyclic 1,2,4-triazol-5 thione analogs were synthesized. The newly synthesized compounds as well as some indole containing thiosemicarbazides, 1,2,4-triazoles and 1,3,4- thiadiazoles, which have been reported previously, were investigated for antimicrobial, antifungal and antiphage activity. Certain thiosemicarbazide derivatives and the corresponding cyclic 1,2,4-triazole analogs showed selective antimicrobial or antifungal activity, while they lack any antiphage activity. Antiphage activity was detected for one compound, bearing the 1,3,4-thiadiazole nucleus. The selectively active compounds cover a wide range of lipophilicity. Structure activity relations show a remarkably similarity in the antimicrobial and antifungal behaviour of the thiosemicarbazides and their cyclic triazo-thien-5-yl analogs, while alpha-naphtyl substitution in the non indolic portion of the molecule is favorable. C5 substitution on the indolic nucleus may also be critical for selective activity. PMID- 10683717 TI - Synthesis, antibacterial, antifungal and anti-HIV evaluation of Schiff and Mannich bases of isatin and its derivatives with triazole. AB - Isatin (indole 2,3-dione) and its 5-chloro and 5-bromo derivatives have been reacted with 3-(4'-pyridyl)-4-amino-5-mercapto-4-(H)-1,2,4-triazole to form Schiff bases and the N-Mannich bases of these compounds were synthesised by reacting them with formaldehyde and several secondary amines. Their chemical structures have been confirmed by means of their IR, 1H-NMR data and by elemental analysis. Investigation of antimicrobial activity of compounds was done by agar dilution method against 27 pathogenic bacteria, 8 pathogenic fungi and anti-HIV activity against replication of HIV-1 (III B) in MT-4 cells. Among the compounds tested 1-(piperidinomethyl) 5-bromo 3-[3'-(4"-pyridyl)-5'-mercapto-4'-(H) 1',2',4'-triazol 4'-yl]imino isatin showed the most favourable antimicrobial activity. PMID- 10683718 TI - Pharmacokinetics and tissue distribution of rifametane, a new 3-azinomethyl rifamycin derivative, in several animal species. AB - Single and repeated dose experiments in mice, rats, dogs and monkeys are reported in this study to assess the pharmacokinetics and tissue distribution of rifametane, a new semi-synthetic rifamycin with the chemical formula 3-[(1 diethylaminoethylidene)azinomethyl]rifamycin SV (CAS 94168-98-6, SPA-S-565). All the kinetic tests were carried out in comparison with known rifamycin derivatives, as rifampicin (CAS 13292-46-1) or rifamycin SV (CAS 6998-60-3). Mice received single i.v. and oral administration of 10 mg/kg of rifametane or of rifampicin and serum samples were obtained up to 96 h after dosing. The two antibiotics showed similar peak of serum concentrations, but rifametane showed a longer half-life and higher AUC values. In an additional experiment, the tissue/serum ratio after the 10 mg/kg oral dose was lower than unity for lungs and kidneys, while the liver/serum ratio exceeded the unity at all sampling times. After 4 weeks of once weekly administration measurable serum and tissue concentrations were observed, and after twice weekly administration for the same time period some blood and tissue accumulation was seen. Rats were treated with a single intravenous injection of 20 mg/kg of rifametane or rifampicin and with single oral or i.m. administration of 60 mg/kg of rifametane or reference standards (rifampicin and rifamycin SV resp.), in two separate trials. The serum half-life of the test antibiotic after i.v. dose was 6 times longer than that of rifampicin and the serum concentrations of rifametane after oral and i.m. doses were higher and longer-lasting than those of the reference compounds. Repeated daily administrations of rifametane at three dose levels (3, 10, 30 mg/kg p.o.) for 4 weeks induced very high serum and liver concentrations. Dogs received a single oral dose of 1.25 mg/kg of rifametane or 2.5 mg/kg of rifampicin. The serum half-life of rifametane resulted 3 times longer than that of rifampicin. Remarkable serum and tissue concentrations were observed after 3-4 weeks of daily oral administration of rifametane at 3, 10, 30 mg/kg dose. Monkeys were given single oral or i.m. administration of 30 mg/kg of rifametane or reference standards (oral rifampicin and i.m. rifamycin SV). The serum concentrations after rifametane were higher and more sustained than those of reference compounds and the half-lives of the test antibiotic were about 2.5 (p.o.) to 6 times (i.m.) longer. The urine excretion of rifametane after a single intravenous dose in rats and a single oral dose in dogs was very low, while rifampicin had a little higher urine concentrations. PMID- 10683719 TI - Effect of quercetin, caffeic acid and caffeic acid phenylethyl ester, solubilized in non-ionic surfactants, on histamine release in vivo and in vitro. AB - A practical hindrance in using many therapeutic agents is their limited solubility in aqueous matrixes. This is usually overcome by incorporating the active compounds in a matrix, with the aid of a non-ionic surfactant. Three water insoluble natural polyphenols with inherent biological activity, quercetin (CAS 117-39-5), caffeic acid and caffeic acid phenylethyl ester, were solubilized in water, with the aid of Tween 80 (an esterified and polyethoxylated derivative of sorbitan), Solutol HS15 (a polyethoxylated derivative of 12-hydroxy-stearic acid), Cremophor RH40 (a ricinoleic acid derivative) or Cremophor EL and the effect of the solubilized polyphenols on histamine release was studied in vitro (mast cells) and in vivo in the rat. In vivo Cremophor EL alone increased, and Tween 80 decreased histamine plasma levels. All four groups injected with solubilized quercetin exhibited a decrease in their plasma histamine levels. Caffeic acid solubilized in Cremophor RH40 decreased histamine levels, too. In vitro Tween 80 increased histamine release in a dose-dependent mode. Quercetin in vitro inhibited histamine release in all solubilizers used. It is concluded that the ability of the studied polyphenols to release histamine is not only depending on the condition of the storage vesicles in the mast cells, but also on the surfactant used to solubilize them. PMID- 10683720 TI - General pharmacology studies on beta-domain deleted recombinant factor VIII. AB - beta-Domain deleted recombinant factor VIII (GC-rAHF), newly developed by Korea Green Cross Co., is a novel therapeutic for hemophiliacs and is currently under clinical evaluation. The general pharmacological properties of this drug were evaluated using mice, rats, guinea pigs and rabbits. Intravenous doses of 5 to 500 IU/kg were assayed in several tests to analyze their effects in vivo on various systems. The effect of the substance under study was also tested in vitro on isolated guinea pig ileum preparations at final concentrations of 5 to 50 IU/kg. The result of this study showed that GC-rAHF did not affect general behavior in the Irwin test. Similarly the drug was not found to affect neither normal body temperature nor the spontaneous activity in mice. In addition, it was not found to induce pharmacologically significant alterations of the cardiovascular and respiratory parameters in rats. No effects were observed either in the pentobarbital sodium-induced sleep-induction time and duration, in writhing test or in the test of pentetrazole-induced convulsion. Finally, the tested drug did not modify the gastrointestinal motility, acetylcholine or histamine-induced contraction of the isolated guinea pig ileum, nor gastric secretion. The results demonstrated that GC-rAHF has no effects on the central nervous, cardiovascular, respiratory and digestive systems in the doses of 5, 50 and 500 IU/kg in vivo and 5, 10, 50 and 100 IU/kg in vitro. PMID- 10683721 TI - Biomedical technology in Franconia. AB - Medical instrumentation and biotechnology business is developing rapidly in Franconia. The universities of Bayreuth, Erlangen-Nurnberg, and Wurzburg hold upper ranks in biomedical extramural funding research. They have a high competence in biomedical research, medical instrumentation, and biotechnology. The association "BioMedTec Franken e.V" has been founded at the beginning of 1999 both to foster the information exchange between universities, industry and politics and to facilitate the establishment of biomedical companies by means of science parks. In the IGZ (Innovation and Foundation Center Nurnberg-Furth Erlangen) 4,500 square meters of space are currently shared by 19 novel companies. Since 1985 60 companies in the IGZ had a total turnover of about 74 Mio Euro. The TGZ (Technologie- und Grunderzentrum) in Wurzburg provides space for 11 companies. For the specific needs of biomedical technology companies further science parks will be set up in the near future. A science park for medical instrumentation will be founded in Erlangen (IZMP, Innovations- und Grunderzentrum fur Medizintechnik und Pharma in der Region Nurnberg, Furch, Erlangen). Furthermore, a Biomedical Technology Center and a Research Center for Bicompatible Materials are to be founded in Wurzburg and Bayreuth, respectively. Several communication platforms (Bayern Innovativ, FORWISS, FTT, KIM, N-TEC VISIT, TBU, WETTI etc.) allow the transfer of local academic research activities to industrial utilization and open new co-operation possibilities. International pharmaceutical companies (Novartis, Nurnberg; Pharmacia Upjohn, Erlangen) are located in Franconia. Central Franconia represents a national focus for medical instrumentation. The Erlangen settlement of the Medical Engineering Section of Siemens employs 4,500 people including approximately 1,000 employees in the Siemens research center. PMID- 10683722 TI - Multiple site mutagenesis with high targeting efficiency in one cloning step. PMID- 10683723 TI - Gene replacement in gram-negative bacteria: the pMAKSAC vectors. PMID- 10683724 TI - Direct observation of GFP gene expression transduced with HSV-1/EBV amplicon vector in unfixed tumor tissue. PMID- 10683725 TI - Dominant positive and negative selection using luciferase, green fluorescent protein and beta-galactosidase reporter gene fusions. PMID- 10683726 TI - Direct reprobing with anti-beta-actin antibody as an internal control for western blotting analysis. PMID- 10683727 TI - Lipopolysaccharide affinity measurement by scintillation proximity assay: application to human heparin binding protein. PMID- 10683728 TI - Suitability of a unique 16S rRNA gene PCR product as an indicator of Gardnerella vaginalis. PMID- 10683729 TI - Expression of TetC fusion proteins from Salmonella in a gaseous environment that models conditions found in mammalian tissues. PMID- 10683731 TI - Transfection of cultured mesenchyme cells isolated from embryonic chick limb buds and facial primordia. PMID- 10683730 TI - Recovery of protein kinases from renatured SDS-polyacrylamide gels for biochemical studies. PMID- 10683732 TI - Fast, qualitative analysis of p53 phosphorylation by protein kinases. PMID- 10683733 TI - Get your bioinformatics on the Web! PMID- 10683734 TI - Photo-cross-linkable oligonucleotide probes for in situ hybridization assays. AB - In situ hybridization techniques have been an important research tool since first introduced 30 years ago, and more recently clinical applications have been expanding greatly. Still, further improvements in the assay sensitivity and protocols that are amenable to routine clinical use are desired. We use a novel photo-cross-linking technology to irreversibly bind short oligonucleotide probes to the target sequence following a hybridization period. The cross-linking agent is incorporated into the backbone of the probe and is activated to react with pyrimidines in the opposite strand by near-UV (300-370 nm) irradiation. By locking the probe to the target, very stringent wash conditions can be used that would otherwise completely remove probes that are hybridized but not cross-linked to the target. Consequently, the probe-specific signal is maximized, while the background signal is minimized to the greatest extent possible with the stringency of the wash. The use of short, photo-cross-linkable probes presents a new strategy for maximizing the sensitivity of probe hybridization or signal amplification-based in situ techniques. PMID- 10683735 TI - Comparison of lipid-mediated and adenoviral gene transfer in human monocyte derived macrophages and COS-7 cells. AB - Lipid-mediated transfection was compared to adenoviral-mediated gene transfer in COS-7 cells as well as human monocyte-derived macrophages (HMDM). For this purpose, we monitored enhanced green fluorescent protein (EGFP) expression by fluorescence microscopy and quantified gene transfer by competitive PCR. Transfection of COS-7 cells with a novel lipid formulation for DNA transfer was highly effective in COS-7 cells. On average, 30% of the cells were fluorescent 48 h after transfection. In HMDM, the same formulation resulted in the expression of EGFP in less than 0.5% of cells. We measured plasmid DNA by quantitative PCR in lipid-transfected macrophages and found that each macrophage contained on average 2 fg of plasmid DNA 24 h after transfection, that is, more than 400 molecules of plasmid DNA entered each cell. Despite the high level of reporter DNA in lipid transfected cells, expression of the fluorescent protein was suppressed in more than 99.5% of the macrophages. We also used adenoviral gene transfer to introduce the foreign DNA into both COS-7 cells and HMDM. Even though the multiplicity of infection was less than 30, expression of EGFP was observed in nearly all COS-7 cells and in more than 80% of HMDM 48 h after transfection. Despite major advances in the field of lipid-mediated transfection of HMDM, the lipid formulations that are available commercially cannot compete with the efficiency of adenoviral gene transfer. PMID- 10683736 TI - Modified differential display technique that eliminates radioactivity and decreases screening time. AB - Several techniques are available that detect variations in gene expression between cellular populations. These include subtractive hybridization (SH), differential colony hybridization (DCH) and mRNA differential display, all based on the analysis of mRNA. The first two techniques, however, are limited because they require large amounts of mRNA for SH or several rounds of screening for DCH. Differential display overcomes both of these limitations. However, the conventional differential display technique is plagued by false positives and is labor intensive. The identification of genes that are truly differentially expressed, therefore, becomes a formidable task. We describe a modified differential display technique that overcomes the limitations of the conventional technique. This new technique eliminates a source of false positives, decreases the time required to screen a set of primers and reduces the use of radioactivity. PMID- 10683737 TI - Specificity-enhanced hot-start PCR: addition of double-stranded DNA fragments adapted to the annealing temperature. AB - A new method to produce hot-start conditions in PCR is described. Short double stranded DNA fragments were found to inhibit the activity of DNA polymerases from Thermus aquaticus and Thermus flavus. This inhibition is not sequence specific, but exclusively dependent on the melting temperature of the fragments as shown by its correlation to their melting curves as measured. This property is exploited by adding fragments of the appropriate length to the PCR mixture during the reaction setup and thereby preventing the DNA polymerases from extending primers annealed nonspecifically at lower than the optimal temperature. By amplifying ten copies of phage lambda DNA in the presence of 2 micrograms of nonspecific DNA, it is shown for three different primer pairs how the melting temperatures of the double-stranded DNA fragments have to be adapted to the cycle profiles to obtain predominantly specific products in the 0.5 microgram range. PMID- 10683738 TI - New protocol for DNA extraction of stool. AB - Present methods for DNA isolation of stool have various limitations such as the amount of stool used, the requirement of lavage fluids or the use of fresh stool. In this paper, a new method is described for the isolation of human nucleic acids from stool, which is independent from the moment of collection. Fecal samples as dry as possible were collected from 75 patients; two grams of stool were mixed with a lysis buffer containing phenol. DNA yields of crude stool were variable and ranged from 9-1686 micrograms/g of feces. With dot blots in 9 of the 75 cases, the human DNA was identified and ranged from 0.06%-46%. In the remaining 66 cases, human genomic DNA was detected by nested PCR, using human K-ras gene amplification as an example. Amplification products were confirmed for human K ras with the exonuclease-amplification coupled capture technique (EXACCT). In conclusion, the developed DNA isolation method can be used for the study of large numbers of stool samples, is independent of the age or method of stool collection and is suitable for large-scale screening studies. PMID- 10683739 TI - Isolation and purification of functional total RNA from woody branches and needles of Sitka and white spruce. AB - The isolation of intact, functional RNA from conifer spp. is not easy, especially from those tissues that are heavily lignified and characterized by a low number of living cells. An efficient procedure for isolating RNA from combined wood and bark tissues of conifers was developed based on a protocol optimized for the extraction of RNA from pollen and one for the isolation of RNA from woody stems. This protocol does not involve the use of phenol, and no ultracentrifugation was required. In addition, the protocol overcame the problems of RNA degradation and low yield due to oxidation by polyphenolics and co-precipitation with polysaccharides, both of which are abundant components in conifer bark tissues. The isolated RNA was of high quality and undegraded as gauged by spectrophotometric readings and electrophoresis in denaturing agarose gels. Quality was further assessed through the subsequent use of the RNA in reverse transcription and RT-PCR, indicating that it could be used for a number of downstream purposes including Northern blot hybridization and cDNA library construction. Using this modified protocol, 80-150 micrograms of RNA was routinely obtained from 1 g of fresh material. This protocol was also used for the isolation of RNA from needles of spruce spp., from which 750-950 micrograms RNA per gram of starting material could routinely be obtained. PMID- 10683740 TI - Cross-contamination limits the use of recycled anion exchange resins for preparing plasmid DNA. PMID- 10683741 TI - Trace contamination following reuse of anion-exchange DNA purification resins. PMID- 10683742 TI - PNA-dependent gene chemistry: stable coupling of peptides and oligonucleotides to plasmid DNA. AB - Two approaches are described for stably conjugating peptides, proteins and oligonucleotides onto plasmid DNA. Both methods use a peptide nucleic acid (PNA) clamp, which binds irreversibly and specifically to a binding site cloned into the plasmid. The first approach uses a biotin-conjugated PNA clamp that can be used to introduce functional biotin groups onto the plasmid to which streptavidin can bind. Atomic force microscopy images of linearized plasmid show streptavidin localized at the predicted PNA binding site on the DNA strand. Peptides and oligonucleotides containing free thiol groups were conjugated to maleimide streptavidin, and these streptavidin conjugates were bound to the biotin-PNA labeled plasmid. In this way, peptides and oligonucleotides could be brought into stable association with the plasmid. A second approach used a maleimide conjugated PNA clamp. Methods are described for conjugating thiolated peptides and oligonucleotides directly to the maleimide-PNA-DNA hybrid. This straightforward technology offers an easy approach to introduce functional groups onto plasmid DNA without disturbing its transcriptional activity. PMID- 10683743 TI - Photoreduction of monoclonal antibodies for conjugation and fragmentation. AB - The conjugation of enzymes, fluorescent or radioactive labels, cross-linkers and other moieties to antibodies is a commonly performed procedure in biochemical research. Using reduced disulphides, conjugation can be an inconvenient, multistep, time- and material-consuming process. We have developed a reduction technique based on UV irradiation, which lacks these drawbacks. Antibodies are irradiated in a sealed vial for a few minutes by a common laboratory UV source in the presence of stannous ions, following the depletion of atmospheric oxygen. The preparation may subsequently be conjugated with thiol-reactive probes such as maleimide derivatives, with no need for any prior purification or concentration. This simple, rapid and effective reduction and conjugation process results in a fully functional immunoglobulin conjugate that can be used for a variety of biochemical applications. PMID- 10683744 TI - Approaches to detecting false positives in yeast two-hybrid systems. AB - While many novel associations predicted by two-hybrid library screens reflect actual biological associations of two proteins in vivo, at times the functional co-relevance of two proteins scored as interacting in the two-hybrid system is unlikely. The reason for this positive score remains obscure, which leads to designating such clones as false positives. After investigating the effect of over-expressing a series of putative false positives in yeast, we determined that expression of some of these clones induces an array of biological effects in yeast, including altered growth rate and cell permeability, that bias perceived activity of LacZ reporters. Based on these observations, we identify four simple strategies that can assist in determining whether a protein is likely to have been selected in a two-hybrid screen because of indirect metabolic effects. PMID- 10683745 TI - Cell-free synthesis and affinity isolation of proteins on a nanomole scale. AB - The performance of conventional cell-free gene expression systems based on the Escherichia coli S30 extract can be significantly improved by using expression vectors that encode viral structural elements known to enhance translation in vivo and to protect mRNA from ribonuclease action. The expression vectors reported here are designed to produce a functionally active protein carrying the Strep-tag oligopeptide at its C-terminus. They can be used in translation, transcription-translation or replication-translation reactions. Depending on its type, the reaction yields up to 40 micrograms per mL, or about 1 nmol of a standard protein. The presence of Strep-tag allows the synthesized protein to be easily isolated on a streptavidin-agarose column under mild conditions and the entire procedure to be completed within one working day. The results show that standard low-cost, cell-free systems can serve for rapid preparation of purified proteins in amounts that can satisfy a number of needs of a research laboratory. PMID- 10683746 TI - Detection of plant genes using a rapid, nonorganic DNA purification method. AB - We have developed a simple procedure for the preparation of plant genomic DNA using FTA paper. Plant leaves were crushed against FTA paper, and the genomic DNA was purified using simple, nonorganic reagents. The 18S rRNA gene and the gene encoding the ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (rbcL) from the chloroplast genome were detected by PCR amplification of DNA on FTA paper. DNA amplification was successful using extracts from 16 dicot and monocot plants. Studies of specific plant extracts revealed that extracts of leaf samples could be collected and stored at room temperature on FTA paper without a decrease in the DNA amplification success rate for more than a month. Both the 18S RNA gene and the rbcL gene were detected in the genomic DNA isolated from various soybean cultivars stored in this manner. Furthermore, by modestly increasing the number of cycles of DNA amplification, we were able to detect the uidA gene in transgenic tobacco and rice leaves as well as a single copy gene linked to the resistance gene of cyst nematode race 3 using genomic DNA isolated on FTA paper. These results demonstrate that genomic DNA isolated using FTA paper can be used for the detection of plant genes, from a wide range of plants with either high or low gene copy number and of either nuclear or cytoplasmic origin. PMID- 10683747 TI - New Cleavase Fragment Length Polymorphism method improves the mutation detection assay. AB - Cleavase Fragment Length Polymorphism (CFLP) analysis is a convenient, accurate and highly sensitive method for the detection and localization of nucleic acid mutations. The assay is well suited for high-throughput screening and can be used to detect mutations in known and unknown nucleic acid samples. A recent improvement in the CFLP assay termed "temperature ramping" or "ramping" is reported here. This procedural improvement eliminates the need for time and temperature optimizations before the actual sample analysis. In this study, we compare the CFLP ramping procedure to the conventional CFLP optimization procedure and demonstrate equal, and in some cases improved, detection of point mutations. With ramping, CFLP reactions are identical for all DNA fragments analyzed, which allows for increased sample throughput, decreased assay time and lower overall cost. PMID- 10683748 TI - [Cardiovascular disorders in Parkinson disease are underrated]. PMID- 10683749 TI - [Orofaciodigital syndrome--a new variant? Psychiatric, neurologic and neuroradiological findings]. AB - Oral-facial-digital (OFD) syndromes are a heterogeneous group of inherited syndromes that have in common anomalies of the face (median cleft lip), the tongue (bifid or lobulated tongue with harmartomas), and the digits (brachydactyly, polydactyly, syndactyly). Due to more or less subtle clinical features, at least seven causally different entities can be identified: 1) OFDS I; 2) OFDS II (Mohr syndrome); 3) OFDS III; 4) OFDS with tibial anomalies (OFDS IV); 5) OFDS V (Thurston syndrome); 6) OFDS VI (Varadi syndrome); and 7) OFDS VII (Whelan syndrome). The neuro-psychiatric clinical observations and MRI findings of a 40 year old woman with a OFD syndrome are described. The observed findings (leukoaraiosis, epilepsy, major depression) in combination with a proven OFD syndrome possibly reflect a new type of OFD syndrome. PMID- 10683750 TI - [German research institute/Max-Planck Institute for psychiatry]. AB - The Deutsche Forschungsanstalt fur Psychiatrie (DFA, German Institute for Psychiatric Research) in Munich was founded in 1917 bel Emil Kraepelin. For a long time it was the only institution in Germany entirely devoted to psychiatric research. Because of its strictly science-oriented and multidisciplinary approach it also became a model for institutions elsewhere. Kraepelin's ideas have certainly had a strong influence on psychiatry in the twentieth century. The fascinating and instructive history of the DFA reflects the central issues and determinants of psychiatric research. First, talented individuals are needed to conduct such research, and there was no lack in this regard. Second, the various topics chosen are dependent on the available methods and resources. And finally, the issues addressed and the ethical standards of the researchers are heavily dependent on the zeitgeist, as is evident in the three epochs of research at the DFA, from 1917 to 1933, from 1933 to 1945, and from the postwar period to the present. With the introduction of molecular biology and neuroimaging techniques into psychiatric research a change in paradigm took place and a new phase of the current epoch began. PMID- 10683751 TI - [A century of German psychiatry (1899-1999)]. AB - The century of German psychiatry between 1899 and 1999 was shaped by creative and catastrophic contrasts. For the beginning and the end these contrasts and the tensions between them can be personalized by the names of Kraepelin and Freud. During the time in between there was the unfolding of the ideas of schizophrenia, classical psychiatry, psychotherapy, structural theories, pharmaco-psychiatry, anthropological psychiatry, social psychiatry and of many other creative theories and methods in manyfold ways. But during the time in between there was also Nazi psychiatry. In historical perspective not only the murdering of countless psychiatric patients was a disaster. Expulsion, intellectual exhaustion, death of many psychiatrists and war damages too had catastrophic consequences. Only seemingly as a paradox Nazi time and its sequels were followed by global acknowledgement for and globalization of German psychiatry. But this period has come to an end too. Resources and man power are available, the path is free for completely new ideas. PMID- 10683752 TI - [Psychiatric classification. Basic idea and development of an ongoing process]. AB - Whereas the present internationally accepted systems appear like a radical break with the psychiatric tradition at first sight, a closer look reveals that there is in fact a continuity between traditional and modern classification systems: The programmatic idea behind their development is still the same as the one that was first formulated by K. Kahlbaum and introduced to psychiatry by E. Kraepelin. And indeed, those changes in modern systems that appear new were developed because the responsible commissions kept in line with this German tradition. The present paper reviews the development of classifications in psychiatry from their beginning to today and underlines the reasons why the true adequacy of current systems will only show in the next millennium. PMID- 10683753 TI - [Comment on R. Thomasius, M. Schmolke, D. Kraus: MDMA ("Ecstasy") use--an overview of psychiatric and medical sequelae]. PMID- 10683754 TI - Vitamin E and ATPases: protection of ATPase activities by vitamin E supplementation in various tissues of hypercholesterolemic rats. AB - It has been shown that the lipid composition of plasma membrane can be modified in vivo by dietary fat. It has also been observed that an increase in the cholesterol content of plasma membranes results in decreased activities of ATPases. In the present study, we evaluated the changes in the activities of ATPases from erythrocytes, hepatocytes, and kidney cortex caused by cholesterol rich diet in rats and subsequently examined the role of vitamin E administration on the cholesterol-induced effects in these tissues. Administration of hypercholesterolemic diet to the rats for 4.5 months, significantly decreased membrane Na(+)-K(+)-ATPase and Ca+2-ATPase activities in comparison to the controls in all tissues studied. Vitamin E supplementation to the hypercholesterolemic rats led to a recovery in membrane ATPase activities. In conclusion, vitamin E supplementation to the rats provided protection against hypercholesterolemic diet-induced impairment of membrane-bound ATPases. PMID- 10683755 TI - Precaecal digestibility of niacin and pantothenic acid from different foods. AB - This study was conducted to investigate the apparent precaecal digestibilities of niacin and pantothenic acid from human nutrient related foods including wheat, coarse whole-meal bread, boiled potatoes and boiled pork and beef. Therefore, pigs were subjected to an end-to-end ileo-rectal anastomosis, so digesta passed straight from ileum to rectum, eliminating endogenous vitamin synthesis. Excreted chyme was collected over 5-days periods, and concentrations of niacin, and pantothenic acid in the food and chyme samples were determined microbiologically. The intestinal bioavailability of niacin and pantothenic acid was affected differently by the food administered. The digestibility values of niacin deriving from the wheat-, potato- and the meat-based meals ranged from 59 to 69%. Wholemeal bread exerted a nutritionally important negative effect on the apparent intestinal availability of dietary niacin relative to the other foods, which averaged by 40%. Food-related differences of the pantothenic acid digestibility values were greater than that observed with niacin. The digestibility values of pantothenic acid from wheat, potatoes and the meat meals ranged between 65 and 81% and were of the order wheat diet > pork diet > potato diet > beef diet, although differences were not statistically significant. The digestibility of pantothenic acid from the coarse wholemeal bread diet was lower than 30%. PMID- 10683756 TI - Lack of effect of dietary chromium supplementation on glucose tolerance, plasma insulin and lipoprotein levels in patients with type 2 diabetes. AB - Chromium is essential for the regulation of insulin action, thereby influencing carbohydrate and lipid metabolism. An uncontrolled pilot study was designed to measure the habitual daily intake of chromium in a group of healthy individuals with type 2 diabetes and to monitor the effect of daily supplementation with high chromium yeast on glucose tolerance, plasma insulin and lipoproteins. Twelve free living adults with type 2 diabetes underwent a glucose tolerance test (GTT) on recruitment, at 4 weeks (after a 7-d duplicate diet collection) and at 12 weeks (following 8 weeks daily supplementation with 100 micrograms of chromium). Urine samples were collected on the day before and the day of each GTT. Blood samples were taken at half hourly intervals for 3 hours during the GTT and the plasma glucose, cholesterol, triglyceride, HDL, LDL and insulin concentration measured. The chromium content of diets and urine samples was determined. Fasting glucose concentrations and glucose area under the curve profiles did not alter significantly post supplementation with the chromium rich yeast. No significant changes in insulin and lipoprotein concentrations were observed. The results of this study do not support the hypothesis that individuals with type 2 diabetes benefit from yeast-based chromium supplements (100 micrograms/day). PMID- 10683757 TI - Effects of capsaicin on biliary free fatty acids in rats. AB - The effects of capsaicin, a major pungent agent of capsicum fruits, on biliary free fatty acids (FFAs) were studied in male rats. Animals were dosed 100 mg/kg capsaicin after the administration of olive oil, and the bile was obtained for 6 hours continuously after dosing with capsaicin for analysis of FFAs using HPLC methods. Capsaicin significantly decreased the total biliary FFA concentration in the animals which had been previously increased by the administration of olive oil. The main FFAs in the bile of control rats are lauric and palmitic acids, followed by linoleic, oleic, stearic and palmitoleic acids. Capsaicin alone decreased the values of these main FFAs. While lauric, palmitic, linoleic, stearic and arachidonic acids were increased significantly by the treatment with olive oil, elevation of these FFAs was inhibited by the treatment with capsaicin. PMID- 10683758 TI - Dietary polyunsaturated fatty acids and plasma butyrylcholinesterase activity in piglets. AB - Diets containing different ratios of n-3:n-6 polyunsaturated fatty acids, were fed to piglets for a period of 10 days. Diets with n-3:n-6 ratios of 0.2 and 0.3 decreased the group mean activity of plasma butyrylcholinesterase when compared with a diet with a ratio of 0.1. PMID- 10683759 TI - Standards for the Ph.D. degree in the molecular biosciences. Recommendations of the Committee on Education of the International Union of Biochemistry and Molecular Biology. PMID- 10683760 TI - How standards for the Ph.D. degree in the molecular biosciences came about. PMID- 10683761 TI - Hypothetical double-helical poly(A) formation in a cell and its possible biological significance. AB - Arguments are presented in favor of capability of poly(A)-tracts of cellular RNA to form double helices in vivo. It is suggested that formation of the double helix in the mRNA poly(A) tall provides the basis for such processes as polyadenylation termination, PAB I synthesis autoregulation, and stabilization of ARE-containing mRNA by ELAV-like proteins. PMID- 10683762 TI - ERK/MAPK pathway is required for changes of cyclin D1 and B1 during phorbol 12 myristate 13-acetate-induced differentiation of K562 cells. AB - Phorbol 12-myristate 13-acetate (PMA)-induced differentiation of human erythroleukemic K562 cells is characterized by growth arrest, morphological change, and expression of megakaryocyte-specific proteins. We examined the possible involvement of cell cycle regulators with PMA-induced growth arrest and megakaryocytic differentiation of K562 cells. The concentrations of cyclin D1 and p21Waf1/Cip1 were dramatically increased, whereas those of cyclin B1 and cdc2 were decreased, by PMA treatment. The concentrations of most cyclin-dependent kinases (Cdk2, Cdk4, and Cdk6), however, were unchanged by PMA treatment. PD98059, a specific inhibitor of MEK1, partially prevented the increase in cyclin D1 caused by PMA and fully reversed the down-regulation of cyclin B1 protein seen in response to PMA treatment. Thus, it is demonstrated here that the PMA-mediated changes of cyclin D1 and B1 are the result of a persistent increase in extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) activity. PMID- 10683763 TI - 5' to 3' single strand DNA exonuclease activity in a preparation of human Ku protein. AB - We describe a novel 5' to 3' single-strand exonuclease activity exhibited by a Ku preparation purified from a human cell line. The enzyme removes 5' single-strand extensions from duplex DNA molecules. The exonuclease and helicase activities respond reciprocally to changes in ATP concentrations: Nuclease activity is inhibited at the ATP concentrations that are optimal for the helicase. The exonuclease activity does not require divalent cations. The potential implications of the exonuclease activity findings for repair of double-strand breaks and recombination processes are discussed. PMID- 10683764 TI - Signature sequences for the galectin-4 subfamily. AB - Galectins are a distinct family of animal lectins that have a cation-independent affinity for beta-galactoside sugars and share characteristic amino acid sequences. The cDNA encoding rabbit bladder galectin-4 has been cloned and sequenced (GenBank accession no. AF091738). The deduced 328 amino acid sequence predicts a multidomain structure consisting of an N-terminal peptide (19 residues) and two carbohydrate recognition domains (130 residues each) connected by a linker region (49 residues). Comparison of rabbit galectin-4 with related proteins reveals that two peptide motifs, M-A-F/Y-V-P-A-P-G-Y-Q-P-T-Y-N-P-T-L-P-Y in the N terminus and A-F-H-F-N-P-R-F-D-G-W-D-K-V-V-F in the first carbohydrate recognition domain are highly conserved in human, pig, rat, and mouse galectin-4 as well as in mouse galectin-6. The two peptide motifs are proposed here as the signature sequences to identify new members of the galectin-4 subfamily. PMID- 10683765 TI - Variation of hepatic methotrexate 7-hydroxylase activity in animals and humans. AB - This study deals with individual and species variations in the converting activity of methotrexate (MTX) to 7-hydroxymethotrexate in animals and humans. When MTX 7-hydroxylase was assayed in six human liver cytosols, a 48-fold range of intersubject variation of the activity was observed. The variations were correlated to the concentrations of aldehyde oxidase activity in human subjects assayed with benzaldehyde as a substrate. Species differences of liver MTX 7 hydroxylase activity were also observed. The activity was highest in rabbits, followed by rats, hamsters, and monkeys but was undetectable in dogs. Strain differences of MTX 7-hydroxylase activity based on aldehyde oxidase activity were also observed in rats and mice. The results suggest that aldehyde oxidase functions as MTX 7-hydroxylase in livers of animals and humans, and the observed differences of MTX 7-hydroxylase activity are due to variations in the amount of aldehyde oxidase present. PMID- 10683766 TI - Single-stranded oligodeoxyribonucleotides are substrates of Fpg protein from Escherichia coli. AB - The interaction of Escherichia coli Fpg protein, which catalyzes excision of several damaged purine bases including 8-oxoguanine (oxoG) from DNA with a set of single- (ss) and double-stranded (ds) 23-mer oligodeoxyribonucleotides (ODNs) containing 8-oxoguanine(s) at various positions, has been investigated. The affinities of different ss ODNs (KM = 0.55-1.3 microM) were shown to be 12-170 times less than those for corresponding ds ODNs (KM = 6-60 nM). Depending on the position of the oxoG within the ODNs, relative initial rates of conversion of ss substrates may be less than, comparable, or greater than those for ds ODNs. The enzyme can remove 5'-terminal oxoG from ODNs only if the 5'-end is phosphorylated. Fpg does not release oxoG residues from the ultimate and penultimate 3'-terminal positions. Duplexes containing two adjacent oxoG are poor substrates for the glycosylase. PMID- 10683767 TI - Fine structure of the human translocation protein 1 (HTP1/TLOC1) gene. AB - We characterized the genomic region corresponding to the human translocation protein 1 (HTP1/TLOC1) cDNA previously reported. An experiment using rapid amplification of cDNA ends revealed that the transcription initiation site was at -12 bp upstream from the translation initiation codon ATG. Using direct sequencing PCR, we determined precise intron/exon boundaries and intron-exon composition of the gene. The gene region spanned approximately 28 kb and was composed of eight exons and seven introns. The lengths of exons and introns range from 48 to > 1707 bp and from 0.25 to 8.2 kb, respectively. The translation initiation codon and the termination codon were located in exons 1 and 8, respectively. The nucleotide sequences of the introns were also determined in the region around the intron/exon boundaries for 63 to 442 bp. All of the sequences around the intron/exon boundaries were consistent with the 5' and 3' consensus sequences for splice junctions of transcribed genes. Putative lariat sequences were identified between -28 and -64 nucleotides from the 3' splice junction for all seven introns. DNA walking experiments revealed a promoter region of 600 bp. The promoter region did not contain an apparent TATA box or a CAT box but did contain Evi-1, GATA, v-Myb, MZF1, and AP-1 binding sites--factors known as regulatory factors on expression of the gene in blood cells. Therefore, this gene may be one such gene. PMID- 10683768 TI - Effects of chlorophyll availability on phycobilisomes in Synechocystis sp. PCC 6803. AB - Inactivation of the chlL gene in Synechocystis sp. PCC 6803 resulted in negligible chlorophyll content when the mutant was grown in darkness. Upon phycocyanin excitation at 580 nm, the 77K fluorescence spectrum of dark-grown cells showed three peaks at 648 nm, 665 nm, and 685 nm, this last being the largest. This reflects the functional presence of major components of phycobilisomes, including phycocyanin, allophycocyanin, and the terminal emitter, and efficient energy transfer between these components. As expected, no fluorescence emission peaks corresponding to chlorophyll in the photosystems were observed. Intact phycobilisomes could be isolated from the dark-grown chlL deletion mutant. However, the phycobilisomes had a lower efficiency of energy transfer than did those isolated from the light-grown mutant, probably because of a decreased phycobilisome stability in the absence of chlorophyll. Exposing the dark-grown chlL-deletion mutant to light triggered the biosynthesis of chlorophyll. For the first 6 h in the light, upon phycocyanin excitation at 580 nm, the 77K fluorescence emission spectrum of greening cells was identical to that of dark-grown cells that lacked significant amounts of chlorophyll. With increased chlorophyll synthesis, gradual energy transfer from phycobilisomes to the two photosystems can be demonstrated. PMID- 10683769 TI - Mechanism of oxidative damage to fish red blood cells by ozone. AB - The present study was conducted to elucidate the adverse effects of ozone exposure on rainbow trout (Oncorhynchus mykiss) red blood cells (RBCs). We evaluated whether hemoglobin (Hb) or Hb-derived free iron could participate in the RBC damage using an in vitro ozone exposure system. Ozone exposure induced hemolysis, formation of methemoglobin, and RBC membrane lipid peroxidation. This RBC damage was not suppressed by the addition of a specific iron chelator (deferoxamine mesilate) to the medium but was suppressed by carbon monoxide (CO) treatment before ozone exposure. Generation of hydrogen peroxide (H2O2) in RBC was observed upon ozone exposure but was significantly suppressed by CO treatment before ozone exposure. Thus the Hb status (i.e., Hb redox condition) and H2O2 generation in RBC should play important roles in mediating RBC damage by ozone exposure. In other words, neither ozone nor its derivative directly attacked from the outside of the cell, but ozone that penetrated through the membrane derived the reactive oxygen species from Hb inside of the cell. PMID- 10683770 TI - Stability of alkyl-dihydroxyacetonephosphate synthase in human control and peroxisomal disorder fibroblasts. AB - Alkyl-dihydroxyacetonephosphate synthase (alkyl-DHAP synthase) is a peroxisomal enzyme that plays a key role in ether phospholipid biosynthesis. To determine the turnover of alkyl-DHAP synthase in several peroxisomal disorders, pulse-chase experiments were performed. In control fibroblasts, mature alkyl-DHAP synthase displayed a half-life of 23 +/- 12 h. In Zellweger syndrome and rhizomelic chondrodysplasia punctata fibroblast cell lines, in which alkyl-DHAP synthase cannot be imported into peroxisomes, the enzyme was mainly detected in its precursor form. This precursor form showed a much shorter half-life, 5 +/- 2 h. In contrast, when the precursor protein accumulated inside the peroxisome of a particular neonatal adrenoleukodystrophy cell line in which processing does not take place, a half-life of 18 +/- 8 h, resembling that of the mature protein in controls, was observed. In a cell line from a patient with a single deficiency in the activity of alkyl-DHAP synthase, the mature form was detected and its radioactivity decreased with a half-life of 16 +/- 7 h. Collectively, these results provide an explanation for the instability of alkyl-DHAP synthase outside its target organelle. Additionally, they indicate that both the precursor and mature form of alkyl-DHAP synthase exhibit considerable intraperoxisomal turnover. PMID- 10683771 TI - Incision size and intraocular lens implantation. PMID- 10683772 TI - Supernormal vision, hypervision, and customized corneal ablation. PMID- 10683773 TI - Interface Elschnig pearl formation with piggyback implantation. PMID- 10683774 TI - IOL registry and errors in A-scan. PMID- 10683775 TI - Drift index to explain patient complaints after PRK. PMID- 10683776 TI - Comments on the combined procedure. PMID- 10683777 TI - Medical control of IOP after cataract surgery. PMID- 10683778 TI - Preventing ocular damage from a cannula. PMID- 10683779 TI - Lidocaine gel for topical anesthesia. PMID- 10683780 TI - Avoiding conjunctival necrosis after periocular depot corticosteroid injection. PMID- 10683781 TI - Simple nucleus cracking technique. PMID- 10683782 TI - Consultation section. Refractive surgical problem. PMID- 10683783 TI - Closed chamber iridodialysis repair using a needle with a distal hole. AB - We describe a closed-system technique of iridodialysis repair using a 22.0 mm, plastic handled, 27 gauge straight needle with a hole 1.0 mm proximal to the tip. The distal hole is suitable for passage of 9-0 or 10-0 mm polypropylene or nylon sutures. The technique was effective in 8 patients with traumatic iridodialysis in the upper and inferior temporal quadrants. PMID- 10683784 TI - In situ tumbling of the AcrySof intraocular lens. AB - The small incision through which foldable acrylic intraocular lenses (IOLs) are implanted does not allow easy explantation of the lens in the event of intraoperative complications. Reversal of the IOL optic during insertion, although rare, can predispose to postoperative complications such as pupillary capture of the IOL, capsule bag distension syndrome, and refractive problems. Explanting the IOL can damage it, the cataract wound, or both. We describe a technique of in situ tumbling of the AcrySof IOL to correct reversed-optic implantation that preserves the integrity of the IOL and anterior segment structures. PMID- 10683785 TI - Surgical prevention of posterior capsule opacification. Part 1: Progress in eliminating this complication of cataract surgery. AB - PURPOSE: To evaluate over almost 2 decades the success of a component of cataract surgery that represents a critical step in reducing the incidence of posterior capsule opacification (PCO); namely, the efficacy of cortical cleanup. SETTING: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Department of Ophthalmology, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: Accessioned from the early 1980s to 1997, 3320 eyes obtained postmortem with posterior chamber intraocular lenses were analyzed with respect to formation of a postoperative Soemmering's ring. This anatomic lesion, the precursor of clinical PCO, represents an important and measurable indication of the quality of cortical cleanup. Its formation was documented using Miyake Apple posterior photographic analysis. RESULTS: The quality and thoroughness of cortical cleanup and overall effectiveness in eliminating retained and/or regenerating cortical cells, as measured by scoring of Soemmering's rings, showed virtually no net change since the early 1980s. The intensity of Soemmering's ring was higher in the most recent specimens than in those in the early 1980s. CONCLUSION: The results indicate that renewed attention to cortical cleanup in cataract surgery is warranted for significant reduction in incidence or the elimination of PCO. More attention to the hydrodissection (cortical cleaving hydrodissection) step of the procedure is likely a practical, immediately implementable, and inexpensive remedy. PMID- 10683786 TI - Surgical prevention of posterior capsule opacification. Part 2: Enhancement of cortical cleanup by focusing on hydrodissection. AB - PURPOSE: To experimentally analyze the role and efficacy of hydrodissection in achieving maximal cortical cleanup. SETTING: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Department of Ophthalmology, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: Phacoemulsification and irrigation/aspiration were performed in 10 pairs of human eyes (20 eyes) obtained postmortem. Ten eyes had previous hydrodissection and 10 eyes, no hydrodissection. The time (seconds) required for complete lens substance removal in each procedure was measured. In addition, a qualitative evaluation of difficulty of surgery was noted. RESULTS: Phacoemulsification required 28.6% less time in eyes with previous hydrodissection than in those without. Irrigation/aspiration time was reduced by 50.9% when hydrodissection was performed. The total time of each procedure required for complete evacuation of the capsular bag was reduced by an average of 37.7% in eyes with hydrodissection. Furthermore, qualitatively the procedure was far easier, less stressful, and caused less posterior capsule stress or rupture when copious hydrodissection was performed. CONCLUSIONS: Hydrodissection enhances the general safety and efficiency of cortical cleanup, especially at 12 o'clock. Hydrodissection is the best available, practical, immediately implementable, and inexpensive means to help remove equatorial E-cells and thus alleviate the incidence of posterior capsule opacification. PMID- 10683787 TI - Surgical prevention of posterior capsule opacification. Part 3: Intraocular lens optic barrier effect as a second line of defense. AB - PURPOSE: To emphasize an important aspect of preventing posterior capsule opacification (PCO), the barrier effect established by the optic of a posterior chamber intraocular lens (PC IOL), and present a new classification regarding capsular bag status after extra-capsular cataract extraction, including phacoemulsification. SETTING: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: This analysis included 150 consecutive eyes obtained postmortem with United States-manufactured PC IOLs including (1) poly(methyl methacrylate), (2) silicone, and (3) hydrophobic acrylic designs that were accessioned in the Center from September 1995 to January 1, 1998. Gross photographs from behind (Miyake-Apple views) were taken and serial histologic sections prepared. RESULTS: Microscopic analysis of the 150 eyes showed that the morphologic appearance of the capsular bag could be grouped into 2 categories: (1) those with little or no evidence of retained cortical material and cells, and (2) those with retained cortical material and cells in which a Soemmering's ring formed. With the latter, when a distinct barricade to cellular migration created by the IOL optic was noted, 2 discrete configurations occurred, depending on the different geometries of the optic components. With a classic biconvex optic with a curved and tapered edge, in many instances some ingrowth of cells proceeded posteriorly around the edge of the IOL optic in the direction of the central axis. With a lens optic that had a squared, truncated, and relatively thick edge, there was often abrupt termination of cells at the peripheral edge of the optic. The posterior capsule subtending the entire optic zone was therefore relatively or totally cell free. CONCLUSIONS: The barrier effect of the IOL optic appears to be of critical importance in retarding ingrowth of cells, functioning as a second line of defense when cortical cleanup is incomplete. Analysis of PC IOLs obtained postmortem showed that a square, truncated optic edge seemed to provide the maximum impediment to cell growth behind the IOL optic. PMID- 10683788 TI - Corneal ablation patterns to correct for spherical aberration in photorefractive keratectomy. AB - PURPOSE: To determine the spherical aberration introduced by photorefractive keratectomy (PRK) and customize ablation patterns to compensate for this aberration and improve post-PRK visual performance. SETTING: Department of Ophthalmology, University of Arizona, Tucson, Arizona, USA. METHODS: Presurgical and postsurgical corneal topography of 16 patients who had PRK with the Summit OmniMed laser were obtained. The data were applied to a schematic eye model, and exact ray tracing was used to determine the introduction of spherical aberration from the procedure. Optimization routines were used to determine the ideal ablation pattern. RESULTS: The magnitude of the spherical aberration introduced into the eyes after PRK increased with the level of attempted correction. The theoretical ideal ablation pattern requires additional flattening of the ablation periphery to avoid the introduction of spherical aberration. CONCLUSIONS: Current PRK ablations introduce spherical aberration into the eye. Modifying the existing ablation algorithms to compensate for spherical aberration may boost postoperative visual performance. PMID- 10683789 TI - Keratoconus evaluation using the Orbscan Topography System. AB - PURPOSE: To evaluate corneal topography in a series of keratoconus patients using the Orbscan Topography System. SETTING: Department of Ophthalmology, Ruprecht Karls-University of Heidelberg, Heidelberg, Germany. METHODS: Seventy-one eyes of 38 patients with keratoconus were evaluated. Quantitative topographic parameters were analyzed with special reference to the central point of the cornea, the apex (the point with maximum reading on the anterior elevation best-fit sphere map), and the thinnest point. Evaluation included location, elevation (compared to a best-fit sphere), pachymetry, tangential curvature, and composite curvature. The mirror-image symmetry between the right and left eyes of a patient was also investigated. RESULTS: Mean patient age was 31.2 years +/- 12.2 (SD). Thirty three patients (86.8%) had bilateral keratoconus and 5 (13.2%), unilateral keratoconus. Most cones (68/71) were located in the inferior temporal quadrant; 3 were above the horizontal meridian. Mean distance between the apex and the thinnest point was 0.917 +/- 0.729 mm (P < .001). The correlations between apex elevation and apex composite curvature and apex tangential curvature were high (r = 0.94 and r = 0.91, respectively; P < .001). In right and left eyes, there was a correlation between the apex and the thinnest point semi-meridians (r = 0.47 and r = 0.65, respectively; P < .05) but not between the radii of the apex and the thinnest point (r = 0.21 and r = 0.24, respectively). CONCLUSIONS: The Orbscan system can provide useful and accurate information in defining the morphology of keratoconus and detecting subtle topographic changes present in early keratoconus. It may also improve the results of contact lens fitting and surgical management. PMID- 10683790 TI - Comprehensive method of analyzing the results of photoastigmatic refractive keratectomy for the treatment of post-cataract myopic anisometropia. AB - PURPOSE: To assess the efficacy, stability, and safety of photoastigmatic refractive keratectomy (PARK) in treating post-cataract myopic anisometropia to restore binocularity and to describe a comprehensive method for analyzing the results of refractive surgery. SETTING: St. Paul's Eye Center, Royal Liverpool Hospital, Liverpool, United Kingdom. METHODS: Nineteen patients (20 eyes) with post-cataract myopic anisometropia were treated with PARK using a VISX Twenty Twenty laser and followed for 12 months. Cataract surgery had been performed between 10 and 144 months (mean 43.4 months) previously. A comprehensive method based on Long's matrix formalism and the vech operator of Harris, in addition to the nearest equivalent sphere and cylinder, was used to analyze the refractive data. RESULTS: The mean preoperative refraction in the post-cataract eyes was 4.79 +1.17 x 0.2 and in the fellow eyes, +0.02 +0.31 x 166. Twelve months after PARK, the postoperative refraction in the post-cataract eyes was -0.90 +0.65 x 2, a significant reduction (P = .15). This postoperative refraction was not significantly different from that in the fellow eye (P = .93). The pretreatment mean uncorrected visual acuity was 0.12. It improved to 0.41 at 12 months, at which time 52% of eyes achieved a visual acuity of 0.5 or better without correction. All patients regained binocularity. At 12 months, 2 eyes (11%) showed clinically unacceptable regression; 1 eye with grade 2 haze lost 1 line of corrected visual acuity. CONCLUSIONS: Photoastigmatic refractive keratectomy reduced post-cataract myopic anisometropia, allowing restoration of binocularity in all patients. Overall, the results in this elderly population with previous ocular surgery, posterior capsule thickening, and macular degeneration are not as satisfactory as those obtained from similar treatment of physiological myopia. Stability and postoperative complications are acceptable. PMID- 10683791 TI - Incision width after phacoemulsification with foldable intraocular lens implantation. AB - PURPOSE: To determine the incision size after insertion of foldable intraocular lenses (IOLs) using both a forceps and injectors. SETTING: Intermountain Ocular Research Center, Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: One hundred patients had phacoemulsification through a clear corneal wound. The incision width was measured before and after IOL implantation. A 3 piece silicone IOL and a 3-piece acrylic IOL (both with an optic diameter of 5.5 mm) were inserted using a forceps. A plate-haptic silicone lens and a 3-piece silicone lens with a 6.0 mm diameter optic were inserted using an injector. RESULTS: The wound size in the group with the 3-piece silicone lens inserted with a forceps enlarged 4.4% (3.23 to 3.38 mm) in the low-power IOL group (15.0 to 20.0 diopters [D]) and 6.2% (3.24 to 3.44 mm) in the high-power IOL group (20.5 to 25.0 D). Similarly, the acrylic IOL wound enlarged 5% (3.21 to 3.37 mm) in the low-power IOL group and 6% (3.25 to 3.44 mm) in the high-power IOL group when a forceps was used. The 3-piece silicone and plate silicone lenses inserted using an injector enlarged the wound 3.2% and 3.3% (3.02 to 3.11 mm and 3.05 to 3.15 mm), respectively. There was no difference in the wound size with higher IOL powers in eyes with injected lenses. CONCLUSIONS: Clear corneal incisions enlarge after insertion of foldable IOLs in a predictable manner, with a forceps-inserted IOL enlarging the wound diameter more than lenses inserted with an injector. The forceps-inserted lens group also showed a difference in wound size related to IOL power. PMID- 10683792 TI - Transconjunctival corneoscleral tunnel "blue line" cataract incision. AB - PURPOSE: To report the technique and astigmatic results of the blue line cataract incision. SETTING: The Buzzard Eye Institute, Las Vegas, Nevada, USA. METHODS: This prospective study included 411 eyes of 271 patients who had cataract extraction by phacoemulsification with a self-sealing 3.0 mm blue line cataract incision. The blue line incision is performed with a diamond knife transconjunctively, 2.0 mm behind the surgical limbus. RESULTS: Mean patient age was 68 years (range 40 to 94 years) and mean preoperative astigmatism, 0.96 diopter (D) +/- 0.78 (SD). Uncorrected visual acuity at 1 day was 20/40 or better in 47% of patients. Mean spherical equivalent was -0.57 +/- 0.78 D at 6 months. Mean postoperative astigmatism measured with a subtraction method was 1.00 +/- 0.84 D at 6 months. Vector analysis showed an induced astigmatism of -0.47 +/- 1.00 D at 1 month, -0.58 +/- 0.81 D at 3 months, and -0.57 +/- 0.99 D at 6 months. No complications such as wound leakage or hyphema occurred. CONCLUSION: The blue line incision combines the efficiency of the clear corneal with the safety of the scleral tunnel cataract incision and appears to be relatively astigmatically neutral. PMID- 10683793 TI - Five year study of astigmatic stability after cataract surgery with intraocular lens implantation: comparison of wound sizes. AB - PURPOSE: To assess the long-term stability of cataract wounds of various lengths. SETTING: Private practice. METHODS: This retrospective study comprised 5 groups of consecutive cataract surgery cases and 1 control group with similar mean ages and wound lengths of 10.0, 6.0, 4.0, 2.0, and 0 (control) mm. Except for the 4.0 mm cases, follow-up was 5 years, with few patients lost during that time. Cases within each group had the same wound position, configuration, and suturing. Refractive data, controlled by keratometry, were collected and analyzed preoperatively and 1 day, 1 and 6 weeks, 3 and 6 months, and 1, 2, 3, 4, and 5 years postoperatively. No sutures were cut. RESULTS: With long-term follow-up, there was a progressive against-the-rule shift in astigmatism. Smaller wounds showed less immediate induced astigmatism. However, except for the unsutured 2.0 mm iridectomy wounds and the control group, all shifted similarly. Data were not available for the 4.0 mm wounds beyond 1 year. CONCLUSION: Wounds were not necessarily "stable" at 6 months. Larger wounds continued to shift years after surgery. Smaller wounds have significant postoperative advantages, but absolute long-term refractive stability may not be one. PMID- 10683794 TI - Combined phacoemulsification, vitrectomy, foreign-body extraction, and intraocular lens implantation. AB - PURPOSE: To assess the outcome of simultaneous phacoemulsification, pars plana vitrectomy, intraocular foreign-body extraction, and intraocular lens (IOL) implantation. SETTING: SSK Ankara Eye Hospital, Department of Vitreoretinal Surgery, Ankara, Turkey. METHODS: Seventeen patients with corneal perforation, intraocular foreign body, vitreous hemorrhage, and lens opacity had simultaneous clear corneal phacoemulsification, pars plana vitrectomy, intraocular foreign body extraction, and IOL implantation. RESULTS: Postoperative complications included massive retinal fibrosis in 2 patients, retinal detachment in 1, and cilioretinal artery occlusion in 1. At a mean follow-up of 15.2 months, best corrected visual acuity improved in the remaining 13 eyes (76%). The IOL was stable in all cases. CONCLUSION: Combined phacoemulsification with IOL implantation and vitreoretinal surgery was safe in selected cases of penetrating ocular trauma resulting from an intraocular foreign body. PMID- 10683795 TI - Perception of Purkinje vessel shadows and foveal granular pattern as a measure of potential visual acuity. AB - PURPOSE: To compare perception of 2 entoptic phenomena, the Purkinje vessel shadows and the foveal granular pattern, as measures of retinal visual acuity using a transscleral illumination technique that bypasses the anterior segment. SETTING: Retinal Vascular Center and General Eye Clinic, Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland, USA. METHODS: Both eyes of 85 patients with clear ocular media, many with retinal disease, were tested for perception of these entoptic phenomena. Also, retinal visual acuity was measured with a Potential Acuity Meter through the current refractive correction. RESULTS: Of 114 eyes with retinal acuity of 20/40 or better, 99% perceived the Purkinje vessel shadows and 86% perceived the foveal granular pattern. Of 45 eyes with retinal acuity of 20/80 or worse, 73% perceived the Purkinje vessel shadows and 4% perceived the foveal granular pattern. CONCLUSION: Perception of the Purkinje vessel shadows does not distinguish between good and poor retinal acuity, whereas nonperception of the vessel shadows strongly suggests poor acuity. Perception of the foveal granular pattern, on the other hand, provides a positive indication of good retinal acuity and will likely prove to be predictive of good vision after removal of significant media opacity. PMID- 10683796 TI - Long-term results of out-of-the-bag intraocular lens implantation. AB - PURPOSE: To evaluate long-term results of out-of-the-bag intraocular lens (IOL) implantation. SETTING: Department of Ophthalmology, Tenri Yorozu Hospital, Nara, Japan. METHODS: This study comprised 22 patients, 13 women and 9 men, who had cataract surgery by phacoemulsification and out-of-the-bag IOL implantation because of a posterior lens capsule rupture. Sixteen patients had in-the-bag IOL implantation in the fellow eye, and these eyes were used as a control group. The IOL's position was determined by ultrasound biomicroscopy (UBM). Anterior chamber flare counts were measured by a laser flare meter. The corneal endothelium was observed by specular microscopy. RESULTS: Mean follow-up after cataract surgery was 35 months +/- 22 (SD). The UBM revealed that in the 19 eyes with sulcus-to sulcus IOL fixation, the optics touched the iris. In 3 eyes, 1 haptic was fixated at the sulcus and the other at the ciliary body. In 2 of these eyes, the optics did not touch the iris. Anterior chamber flare counts in eyes with sulcus-to sulcus IOL fixation were significantly higher than in eyes with in-the-bag or sulcus-to-ciliary-body fixation (P < .05). There were no statistical differences in corneal endothelial cell counts based on haptic placement. CONCLUSION: Rubbing between the IOL optic and iris seems to contribute to the high flare counts in eyes with a sulcus-to-sulcus IOL fixation. A larger haptic angle may be needed to prevent contact between the iris and IOL optic in such cases. PMID- 10683797 TI - Intraocular pressure after small incision cataract surgery with Healon5 and Viscoat. AB - PURPOSE: To evaluate the effect of Healon5 (sodium hyaluronate) and Viscoat (sodium chondroitin sulfate-sodium hyaluronate) on intraocular pressure (IOP) after bilateral small incision cataract surgery. SETTING: Department of Ophthalmology, University of Vienna, Vienna, Austria. METHODS: This prospective randomized study comprised 70 eyes of 35 consecutive patients with age-related cataract in both eyes scheduled for bilateral small incision cataract surgery. The patients were randomly assigned to receive Healon5 or Viscoat during cataract surgery in the first eye. The second eye received the other viscoelastic substance. Cataract surgery was performed in an identical fashion in both eyes, with a temporal 3.5 mm sutureless posterior limbal incision, phacoemulsification, and implantation of a foldable silicone intraocular lens. The IOP was measured preoperatively and 6 hours, 20 to 24 hours, and 1 week postoperatively. RESULTS: At 6 hours after surgery, the mean IOP increased by 5.2 mm Hg +/- 5.3 (SD) in the Healon5 group (P < .0001) and by 10.1 +/- 8.7 mm Hg in the Viscoat group (P < .0001). The increase was significantly higher in the Viscoat group than in the Healon5 group (P = .0016). Intraocular pressure spikes of 30 mm Hg or more occurred in 2 eyes in the Healon5 group and in 10 eyes in the Viscoat group (P = .0112). Twenty to 24 hours and 1 week postoperatively, the mean IOP in the 2 groups was not statistically different. CONCLUSIONS: Viscoat caused a significantly higher IOP increase and significantly more IOP spikes than Healon5 in the early period after small incision cataract surgery. PMID- 10683798 TI - Effect of Healon and Viscoat on outflow facility in human cadaver eyes. AB - PURPOSE: To compare the acute effects of Healon (sodium hyaluronate) and Viscoat (sodium chondroitin sulfate-sodium hyaluronate) on outflow facility in human cadaver eyes and determine which viscoelastic agent is least likely to cause an intraocular pressure (IOP) spike after cataract surgery. SETTING: The Glaucoma Research Lab, University of Toronto, Ontario, Canada. METHODS: In this prospective paired study, 15 pairs of human cadaver eyes were used. Following the construction of a 3.0 mm scleral tunnel, 0.25 cc of Healon was injected into the anterior chamber of 1 eye and 0.25 cc of Viscoat was injected into the contralateral eye. The viscoelastic agents were removed from both eyes in a standardized fashion and the scleral tunnels closed. The eyes were then perfused at a constant IOP of 8.0 mm Hg, corresponding to 16.0 mm Hg in vivo. Outflow facility (microL/minute [min]/mm Hg) was recorded every 15 minutes for 24 hours using standard methods. RESULTS: Outflow facility in the Viscoat-treated eyes decreased appreciably for the first 3 hours, then recovered somewhat after 12 hours; facility in the Healon-treated eyes showed less of an overall decrease. Over the 24 hour perfusion period, mean outflow facility was 0.037 microL/min/mm Hg +/- 0.015 (SD) in the Viscoat-treated eyes and 0.060 +/- 0.012 microL/min/mm Hg in the Healon-treated eyes. Healon reduced outflow facility significantly less than Viscoat between 3.25 and 10.50 hours postoperatively (P < .05, 2-tailed t test). CONCLUSIONS: Healon reduced outflow facility less than Viscoat between 3.25 and 10.50 hours postoperatively. PMID- 10683799 TI - Ultrasonic transmission in viscoelastic substances. AB - PURPOSE: To study the propagation of ultrasonic shock waves in viscoelastic agents and the resulting corneal load. SETTING: University Siegen, Institute for Mechanics and Control Engineering, Siegen, Germany. METHODS: The anterior chamber of a manufactured artificial eye was constructed according to anatomic dimensions. Three openings were drilled--for the phaco tip, for the exchange of a viscoelastic agent or water, and for the shock-wave sensor. The sensor was fixed to the area corresponding to the corneal apex. The sensor signal was analyzed using a direct oscilloscope that measured the amplitude reaching the corneal apex. Shock-wave propagation in several viscoelastic agents was compared with that in balanced salt solution. RESULTS: In hydroxypropyl methylcellulose, the shock wave was amplified or influenced slightly. In hyaluronic-acid preparations, acoustic dampening occurred. CONCLUSION: Removal of hyaluronic-acid derivatives prior to phacoemulsification is not necessary. PMID- 10683800 TI - Detection of integrins in cataract lens epithelial cells. AB - PURPOSE: To detect the expression of integrin subunits in lens epithelial cells (LECs) of human cataracts. SETTING: Research Laboratory, International Intraocular Implant Training Centre, Tianjin Medical University, China. METHODS: The circular sections of the anterior capsules with attached LECs were obtained during cataract surgery from 100 patients. The LECs were stained with an avidin biotin-complex immunohistochemical technique using 5 monoclonal antibodies specific for alpha subunits 2, 3, 5 and beta subunits 1, 2. RESULTS: All integrin subunits studied were found to varying degrees in specimens. The positive percentages were 70%, 65%, 75%, 70%, and 80%, respectively. CONCLUSION: Integrin subunits were present in LECs of human cataracts. These molecules may serve in the adhesion of LECs to the lens capsule and play a role in cell-posterior capsule interaction after cataract surgery. PMID- 10683801 TI - Corneal ectasia detected after laser in situ keratomileusis for correction of less than -12 diopters of myopia. AB - We report 2 cases of corneal ectasia detected after laser in situ keratomileusis (LASIK) for the correction of less than -12.0 diopters (D) of myopia. Patients were evaluated before and after LASIK by corneal topography and pachymetry. After treatment, visual acuity temporarily improved but was followed by visual impairment, with corneal ectasia detected by topography. There may be a risk of corneal ectasia after LASIK in cases of myopia of less than -12.0 D. Despite thelow incidence, we recommend that LASIK be restricted to cases in which more than half the original corneal thickness and more than 250 microns of the stromal bed can be preserved. Careful examination, including preoperative serial topographic evaluation and measurement of posterior stromal thickness, should be performed to improve the quality and predictability of corneal refractive surgery. PMID- 10683802 TI - Phototherapeutic keratectomy of a corneal scar due to presumed infection after photorefractive keratectomy. AB - This case involves a 25-year-old patient who suffered from corneal ulceration several days after photorefractive keratectomy (PRK). A central scar developed, resulting in discomfort and reduction in visual acuity. Four months later, the scar was treated by phototherapeutic keratectomy (PTK) (25 microns depth, 5 mm ablation zone). Some scar tissue was left, but it cleared slowly and steadily over the next few years. The induced hyperopia decreased from 5.00 to 1.37 diopters spherical equivalent within 28 months postoperatively. Best corrected visual acuity increased from 20/60 preoperatively to 20/20 at 28 months postoperatively. Surgeons can encourage patients with postinfectious scars after PRK to try at least 1 PTK treatment. PMID- 10683803 TI - Silicone oil removal from a silicone intraocular lens with perfluorohexyloctane. AB - Silicone oil can be easily dissolved in vivo from silicone intraocular lenses by perfluorohexyloctane. Using specular microscopy, we examined the corneal endothelium in a patient with silicone lens implantation immediately after cataract surgery and 5 weeks postoperatively. There were no signs of acute or chronic toxicity to the endothelium. PMID- 10683804 TI - Calcium absorption from the ingestion of coral-derived calcium by humans. AB - Recent dietary life involves frequent opportunities for the ingestion of purified, processed food products and preserved foods, and it has been pointed out that the current dietary mineral intake strongly tends toward nutritional imbalance. The Ryukyu Islands yield coral which contains calcium and magnesium in a content ratio of about 2 to 1, with their approximate contents of 20 and 10%, respectively. In this report, the calcium absorption from the ingestion of crackers into which the coral powder was incorporated (coral-added crackers) and that from ingestion of calcium carbonate-added crackers was comparatively assessed. Twelve healthy adult volunteers (6 men and 6 women) ingested coral added crackers (calcium content: 525 mg) and calcium carbonate-added crackers (ditto) once each alternately on a cross-over design with a wash-out period of 3 d between the regimens. The study also included controls receiving neither cracker. The degree of intestinal absorption of calcium from coral-added crackers and that from calcium carbonate-added crackers was evaluated in terms of increment in urinary calcium excretion per dL of glomerular filtrate (GF) (difference between coral calcium and calcium carbonate) and increase in urinary calcium excretion per milligram creatinine (difference from control value). The increment in urinary calcium excretion per dL of GF during the latter half of the observation period after the ingestion of coral-added crackers was significantly greater than that during the latter half of the observation period after ingestion of calcium carbonate-added crackers (p = 0.039, paired t-test). A significant difference (from control value) in the increase of urinary calcium excretion per milligram creatinine was also observed (p = 0.0008). The present data, though from a relatively few study subjects, suggest that the calcium of coral origin is better absorbed from the intestine than calcium of calcium carbonate origin on the average. PMID- 10683805 TI - Foods contributing to absolute intake and variance in intake of selected vitamins, minerals and dietary fiber in middle-aged Japanese. AB - Using 351 one-day weighted diet records, we selected foods providing vitamins, minerals and dietary fiber according to contribution analysis (CA) and multiple regression analysis (MRA). Vitamin C was supplied by various vegetables and fruits, and carotene was specifically derived from green-yellow vegetables based on MRA as well as CA. Vitamin A was provided by green-yellow vegetables, fruits, chicken egg and milk (whole) according to CA; whereas chicken liver and pork liver were major sources according to MRA. Vitamin E was mainly of vegetable origin as determined by CA, and largely of spinach, safflower oil and pumpkin as determined by MRA. Vitamin D was mainly derived from chicken egg, fish and mushroom based on CA, and particularly from fish based on MRA. Calcium was supplied by milk (whole), soy products and chicken egg as determined by CA; while milk, tofu and various small fishes were the main contributors to variance. Magnesium was provided by soy products, well-milled rice and spinach according to both analyses, and iron by chicken egg, spinach and soy products. Zinc was largely derived from well-milled rice, followed by chicken egg and milk (whole) as determined by CA, and copper was provided by well-milled rice, soy and wheat products. Dietary fiber was supplied by vegetable sources, whether water soluble or insoluble, based on both analytic methods. PMID- 10683806 TI - Oral health and nutritional status in Egyptian elderly. AB - A cross-sectional sample of 253 ambulatory elderly Egyptians (99 males and 154 females) of minimal age, 60 y, living in the metropolitan Cairo area was investigated. Nutritional status assessment was based on body weight, mid-upper arm circumference and skinfold thickness. Additionally, three nutritional indices, namely body mass index, index of adiposity and index of muscularity, were computed. The oral cavity was examined clinically to evaluate the number and health condition of teeth present. Results showed that the anthropometric nutritional indicators were higher than expected, however, they decreased by age in both sexes. In addition, the values of body mass index and index of adiposity in females were significantly higher than in males, while the reverse was observed in regards to muscularity index (p < 0.05). The three nutritional indices in dentate subjects were slightly higher than those in edentulous ones. Moreover, within the dentate groups there was a tendency for increased values of these indices by the increasing number of healthy teeth. PMID- 10683807 TI - Effect of timing of meal intake after squat exercise training on bone formation in the rat hindlimb. AB - We hypothesized that bone acquisition was affected by the timing of meal intake after resistance exercise training. This was based on the following previous results: 1) Nutrient intake right after exercise resulted in an increase in muscle mass and a decrease in abdominal fat mass as well as muscle protein synthesis when compared to the intake of a meal later after the exercise; and 2) body composition has been proposed to be a good predictor of bone mass. To substantiate our hypothesis, 20 male rats were assigned to either a group fed a meal right after squat exercise (R) or a group fed a meal 4 h after the exercise (L). The 10-wk training program consisted of approximately 70% of one repetition maximum for each animal, 15 repetitions per set, 10 sets per day, 3 d per week. As a result, hindlimb muscle mass in the R group was greater (p < 0.05) than that in the L group and abdominal fat mass was less (p < 0.01) in the R group as compared to the L group, regardless of there being no significant difference in body weight between the groups. Bone volume in the tibia (p < 0.01) and femur (p < 0.05) were both significantly greater in the R group than in the L group. Bone mineral content index (BMCI) and bone mineral density index (BMDI) in the tibia of the R group were significantly (p < 0.05) greater than the corresponding values of the L group. The greater BMCI and BMDI in the tibia were positively and significantly (p < 0.05) related with hindlimb muscle mass, but not with abdominal fat mass. There was no significant difference in BMCI and BMDI in the femur between the groups. These results suggest that the R regimen may contribute to increased bone acquisition in the tibia as compared to the L regimen, and this effect is partly due to the enlargement of muscle mass in the R group as compared to the L group. PMID- 10683808 TI - Effects of long-term consumption of high doses of fish oil concentrates on clinical parameters in male and female rats. AB - Many studies suggest that a diet supplemented with fish oil concentrates (FOCs) may provide protection against cardiovascular and other diseases. The possible harmful effects of long-term consumption of high doses of FOCs, however, have not been adequately investigated. Corn oil, fish oil (MaxEPA) and various mixtures of the oils were administered by gavage to 120 male and 120 female rats, 5 d/wk for 13 wk at the rate of 5 mL/kg/d. Although MaxEPA had no effect on prothrombin time or activated partial thromboplastin time, it caused a statistically significant diminution of the total serum cholesterol level. Correlations between relative liver and spleen weights and dose levels were positive but a negative correlation was found between dose levels and serum vitamin E concentration. In female rats, the negative correlations between dose levels and serum iron and triglyceride levels were highly significant. The pathology data showed no remarkable lesions in any of the tissues examined. Results of this study suggest that long-term consumption of high levels of FOCs in rats may reduce serum cholesterol and triglycerides and adversely affect serum iron level and relative liver weight in female rats and relative spleen weights in both sexes. PMID- 10683809 TI - Effect of a new amino acid solution on nutritional status and nitrogen metabolism in rats with chronic renal failure undergoing hyperalimentation. AB - We studied the effects of the new amino acid solution MRX-III on the nutritional status and nitrogen metabolism of rats with chronic renal failure (CRF) in comparison with those of a general amino acid solution (MPR-F). The essential amino acids/non-essential amino acids ratio was 3.21 for MRX-III and 1.09 for MPR F. Rats with CRF, induced by 7/8 renal ablation, were divided into 6 groups of 8 rats each receiving total parenteral nutrition (TPN) containing MRX-III or MPR-F at a non-protein calorie/nitrogen ratio (Cal/N) of 300, 600 or 900 for 7 d. The rats were infused with test solutions containing the same amounts of non-protein calories. The cumulative nitrogen balance, as a nutritional index, in the MRX-III group was significantly higher than that in the MPR-F group at the Cal/N of 600 or 900, and the plasma albumin level at the Cal/N of 300. The plasma transferrin levels at the Cal/N of 900 in the MRX-III groups were significantly higher than those in the corresponding MPR-F groups. At all Cal/N, the MRX-III groups showed low levels of blood urea nitrogen and urinary excretion of ammonia and urea nitrogen as compared with the MPR-F groups at the same Cal/N. The plasma amino acid concentration profiles in the MRX-III groups after TPN showed greater similarity to that in the Normal group as compared with the profiles in the corresponding MPR-F groups. No aggravation of renal failure was observed in any TPN groups during TPN. These results indicate that, in rats with CRF undergoing hyperalimentation, the effects of MRX-III on the nutritional status and nitrogen metabolism are superior to those of the general amino acid solution, MPR-F. It is suggested that MRX-III could safely provide adequate amounts of nitrogen during hyperalimentation. PMID- 10683810 TI - Effect of the fat/carbohydrate ratio in the diet on obesity and oral glucose tolerance in C57BL/6J mice. AB - To study whether consumed dietary fat has a linear relationship or a threshold with glycemic controls, female C57BL/6J mice were fed different levels of a safflower oil (10, 20, 30, 40, 50, and 60% of total energy) diet ad libitum for 15 wk. Food intake, body weight, parametrial white adipose tissue (WAT) and liver weight were measured, and oral glucose tolerance tests were conducted. Although there was no significant difference in average energy intake, graded increments of safflower oil resulted in graded deterioration of glucose tolerance during 5 and 12-wk feeding, and deterioration of glucose tolerance was more manifested after 12-wk feeding as compared to 5-wk feeding. After 12-wk feeding, a significant deterioration of glucose tolerance was observed in diets of more than 40% fat. Graded increments of body weight and WAT weight were observed, and their weight increases were manifested in diets of more than 30% fat. These data indicated that the amount of dietary fat had an almost linear relationship with glucose tolerance, and significant differences were observed in mice fed diets more of than 40% fat. PMID- 10683811 TI - Comparison of various phosphate salts as the dietary phosphorus source on nephrocalcinosis and kidney function in rats. AB - The effects of various phosphate salts as the dietary phosphorus sources on the development of nephrocalcinosis and kidney function were examined in rats fed diets containing monophosphate salts (sodium dihydrogenphosphate, NaH2PO4, or potassium dihydrogenphosphate, KH2PO4) or polyphosphate salts (sodium tripolyphosphate, Na5P3O10, or potassium tripolyphosphate, K5P3O10), at levels representing normal phosphorus (normal phosphorus diet) or high phosphorus (high phosphorus diet) contents for 21 d. High phosphorus diet-feeding increased the kidney calcium and phosphorus concentrations. Kidney calcium and phosphorus concentrations were higher in rats fed the high phosphorus diet containing Na5P3O10 or K5P3O10 than in rats fed the high phosphorus diet containing NaH2PO4 or KH2PO4. Nephrocalcinosis was observed in all rats fed a high phosphorus diet, and the degree of nephrocalcinosis was more severe in rats fed Na5P3O10 or K5P3O10 than in rats fed NaH2PO4 or KH2PO4. In rats fed the high phosphorus diet, creatinine clearance was higher in rats fed Na5P3O10 or K5P3O10 than in rats fed NaH2PO4 or KH2PO4. In rats fed Na5P3O10 or K5P3O10, urinary albumin excretion and N-acetyl-beta-D-glucosaminidase (NAG) activity in the urine were increased in rats fed the high phosphorus diet. These were higher in rats fed the high phosphorus diet containing Na5P3O10 than in rats fed the high phosphorus diet containing NaH2PO4 or KH2PO4. This study observed that the development of nephrocalcinosis and kidney function in rats fed the high phosphorus diet was influenced by the difference in monophosphate or polyphosphate salts provided as the dietary phosphorus source, while the effects of sodium and potassium salts were not evident. We suggest that the development of nephrocalcinosis and kidney function in rats fed a high phosphorus diet was altered depending on the form of phosphate salts provided as the dietary source of phosphorus. Additionally, the development of nephrocalcinosis and diminished kidney function in rats fed the high phosphorus diet was more severe for polyphosphate salts as compared to monophosphate salts. PMID- 10683812 TI - Ultrasonography evaluation of abdominal fat in live rats. AB - We have developed a new noninvasive method of estimating abdominal fat volume in live rats using ultrasonography. By this method, cross sections of perirenal (retroperitoneal) fat tissue, which is an abdominal fat, at the renal vein level could be identified and the area determined. The perirenal fat in Wistar rats (wide body weight range, 111.4 to 497.3 g; limited range, 300.1 to 337.9 g) measured by ultrasonography was compared with the actual fat tissue weight. The cross-sectional area of perirenal fat tissue was significantly correlated to the actual whole tissue weight. Using this procedure, we examined the changes of perirenal fat stores during fasting. Consequently, the cross-sectional area of perirenal fat and its actual weight decreased in parallel. Total body electrical conductivity (TOBEC) is currently used to measure fat-free mass (FFM) and indirectly predicts total body fat mass of live laboratory animals. The body fat distribution, that is, the location of adipose tissue in the abdominal region, is closely associated with obesity-related diseases. Therefore, it is important to focus not only on the accumulation of total body fat, but also on that of abdominal fat. The present ultrasonographic method is considered to be useful for repeated noninvasive measurement of abdominal fat in the live rat. PMID- 10683813 TI - The correlation between feed-intake cycle and nutritional zinc-deficient status in rats. AB - The characteristic cyclic variation in feed intake of rats fed a Zn-deficient diet (Mills et al, Am J Clin Nutr 22: 1240-1249 (1969)) followed a Cosinor curve, as determined by computer analysis (Tamaki et al, Br J Nutr 73: 711-722 (1995)). The values of amplitude for the feed-intake cycle had a positive correlation to their own day-to-day variations and to the correlation value of their own simulated cycles (r2 = 0.764, df = 50, p < 0.001 and r2 = 0.682, df = 50, p < 0.001, respectively). The cyclic variation in feed intake was accompanied by a cyclic variation in body-weight change in rats fed the Zn-deficient diet, and cyclic variation in body-weight change occurred similarly in pair-fed control rats. There were no differences in the mesors of body-weight change cycles of Zn deficient rats and pair-fed control rats (Zn-deficient rats: 2.5 +/- 1.0 g/d, pair-fed rats: 2.8 +/- 1.0 g/d, mean +/- SD, df = 18, t = -0.674, ND). Rats fed the Zn-deficient diet were given different amounts of Zn supplementation by daily subcutaneous injection. The amplitude of the feed-intake cycle was decreased with increasing Zn supplementation (r2 = 0.919, df = 5, p < 0.001). The concentration of Zn for the appearance of the feed-intake cycle was estimated to be 71.6 +/- 6.6 micrograms/d per rat. The Zn level in the serum showed a significant decrease in the Zn-deficient diet groups, but the supplement of Zn did not vary in the Zn deficient rats injected with up to 47.3 micrograms/d per rat. From these results, an analysis of the feed-intake cycle allowed us to estimate the quantitative Zn deficient status of rats. PMID- 10683814 TI - Depletion of dietary n-3 fatty acid affects the level of cyclic AMP in rat hippocampus. AB - Prolonged depletion of dietary n-3 fatty acid induces a neurological disturbance. To ascertain the deficit of neurotransmission at the time of n-3 deficiency, the concentrations of cAMP and inositol triphosphate, and the activities of protein kinases A and C were examined in vitro in rat hippocampus. Furthermore, the saturation binding study of [3H]quinuclidinyl benzilate, a specific antagonist to muscarinic cholinergic receptor, was performed. Rats were fed a safflower oil diet as the deficient group and a soybean oil diet as the control group. Hippocampi were obtained from rats in the 3rd generation in the deficient group and in the 2nd generation in the control group. Dietary effect was not observed in the parameters except for the concentration of cAMP, which was significantly higher in the deficient group than in the control group. PMID- 10683816 TI - The effect of docosahexaenoic acid on plasma catecholamine concentrations and glucose tolerance during long-lasting psychological stress: a double-blind placebo-controlled study. AB - We previously found that docosahexaenoic acid (DHA) intake prevented aggression from increasing at times of mental stress. In the present study, we investigated whether DHA intake modified the plasma catecholamines and cortisol of medical students during a 9-wk period of final exams. We also investigated the effects of DHA intake on a 75 g oral glucose tolerance test (oGTT). Fourteen medical students participated in the present study. They were randomly allocated to either control or DHA group in a double-blind manner. Subjects in the control group (4 males and 3 females) took 10 control capsules/d, each capsule containing 280 mg of mixed plant oil, and those in the DHA group (4 males and 3 females) took 10 DHA capsules/d containing 1.5 g DHA for 9 wk, during which subjects underwent more than 20 stressful final exams. At the start and end of the study, plasma catecholamines (epinephrine, norepinephrine (NE) and dopamine) and cortisol were measured; a 75 g oGTT was also performed. There were no intra- or intergroup differences in plasma glucose concentrations. However, NE concentrations were significantly reduced after DHA administration (-31%, p < 0.03). The other catecholamines and cortisol did not change significantly. The plasma ratio of epinephrine to NE increased in every DHA subject (+78%, p < 0.02), and intergroup differences were significant (p < 0.03). We conclude that these effects of DHA may be applied to people under long-lasting psychological stress to prevent stress-related diseases. PMID- 10683815 TI - Allylthiamindisulfide and related compounds enhance thermogenesis with increasing noradrenaline and adrenaline secretion in rats. AB - The effects of allylthiamindisulfide, an allyl derivative of thiamin, and related compounds on thermogenesis were investigated by measuring noradrenaline and adrenaline secretion and the temperatures of interscapular brown adipose tissue (IBAT) and rectum in rats. In Experiment 1, the effects of the administrations of allylthiamindisulfide and related compounds on noradrenaline and adrenaline secretion were evaluated as compared to thiamin in anesthetized rats. The administration of allylthiamindisulfide significantly increased the plasma concentrations of noradrenaline and adrenaline. These increases were dose dependent, while that of thiamin was not. Four synthetic compounds related to allylthiamindisulfide also increased the plasma adrenaline and noradrenaline concentrations. In Experiment 2, the effects of allylthiamindisulfide on thermogenesis were investigated by the direct measurement of temperatures in the IBAT and rectum in anesthetized rats, and compared to the effects induced by thiamin and diallyldisulfide. The temperatures in the IBAT and rectum were significantly increased by the administration of allylthiamindisulfide and diallyldisulfide, while there was no significant increase as the result of thiamin administration. These results suggest that allylthiamindisulfide and related compounds enhance thermogenesis by increasing noradrenaline and adrenaline secretion in rats. PMID- 10683817 TI - Effects of high-fat diet intake on glucose uptake in central and peripheral tissues of non-obese rats. AB - We previously demonstrated that plasma glucose concentration was higher while plasma insulin concentration was lower in rats fed a high-fat diet. In the present study, we examined the effects of high-fat diet on glucose uptake in central and peripheral tissues in non-obese rats. Forty male Sprague-Dawley rats were fed high- or low-fat diets for 4 wk. Body weight and body fat accumulation were not different between the two diet groups after 4 wk. Glucose uptake in the skeletal muscles and adipose tissues, estimated by the 2-deoxy-D-glucose method, was lower in the rats fed the high-fat diet than that in the rats fed the low-fat diet, whereas uptake in the liver and pancreas did not differ between the two groups. Glucose uptake in the hypothalamus and cortex was higher in the high-fat diet group as compared with that in the low-fat diet group. These results suggest that increased plasma glucose levels in rats fed the high-fat diet were caused by a decrease in glucose uptake in the skeletal muscles and adipose tissues. Reduced plasma insulin level in the high fat diet group with no difference in glucose uptake in the pancreas may be due to increased sympathetic activity in the pancreas resulting from the increased glucose uptake in the brain regions involved in autonomic functions. PMID- 10683818 TI - Towards the unknown. PMID- 10683819 TI - Unipolar brush cells in marmoset cerebellum and cochlear nuclei express calbindin. AB - Unipolar brush cells (UBCs) are excitatory neurons in the mammalian cerebellum and cochlear nuclei (CN), including the CN of primates, as shown only recently. UBCs are readily identified by their expression of the calcium-binding protein calretinin (CR), except for those of the primate CN that hardly immunostain for CR. The present findings corroborate the existence of UBCs in the CN of a primate, Callithrix. Furthermore, evidence is presented for UBCs, in the cerebellum and a small subpopulation of UBCs in the CN of Callithrix to express the calcium-binding protein calbindin (CB). This may be unique for Callithrix as CB-expressing UBCs have not been recognized in any other mammal. Presence of CB but not CR in UBCs of the Callithrix CN may indicate a certain interchangeability between these two calcium-binding proteins. PMID- 10683820 TI - Pattern of distribution and co-localization of NOS and ATP in the myenteric plexus of human fetal stomach and intestine. AB - The pattern of distribution and co-localization of nitric oxide synthase (NOS) and quinacrine fluorescence (indicative of vesicular adenosine 5'-triphosphate, ATP), and co-localization of NADPH-diaphorase (NADPH-d) activity and NOS immunoreactivity in the myenteric plexus of pre-term human fetal (6-17 weeks of gestation) stomach and small intestine was examined using immunohistochemical and histochemical techniques. In all stages of gestation investigated, NOS immunoreactive and NADPH-d-reactive myenteric neurons and nerve fibres were seen in the fetal intestine and stomach. However, in fetuses of 6-10 weeks of gestation, only 15% of the NADPH-d-positive myenteric neurons were NOS immunoreactive, whereas a 100% co-localization was found in samples of 12-17 weeks of gestation. Quinacrine fluorescent myenteric neurons and nerve fibres were found only in the fetal intestine of 12-17 weeks of gestation, of which 25% of the NADPH-d-positive myenteric neurons in these samples were quinacrine fluorescent. These findings demonstrate the presence and co-localization of markers for nitric oxide (NO)- and ATP-utilizing myenteric neurons and nerve fibres in the early stages of gestation, suggesting possible co-transmitter and/or trophic roles of ATP and NO in the process of development and maturity of human myenteric neurons. In addition, the fact that only a small percentage of NADPH-d-reactive myenteric neurons express NOS immunoreactivity at 6-10 weeks of gestation confirms that NADPH-d-reactivity does not always represent NOS activity. PMID- 10683821 TI - Immunolocalization of NAIP in the human brain and spinal cord. AB - The neuronal apoptosis inhibitory protein (NAIP) is known to have anti-apoptotic functions, and its gene is often mutated in severe cases of spinal muscular atrophy (SMA), a disease characterized by motor neuron degeneration. In this study, we examined the distribution of the endogenous NAIP protein in normal human spinal cord and brain tissue by using a polyclonal antibody against NAIP. Immunohistochemical staining demonstrated that NAIP is strongly expressed in anterior horn and motor cortex neurons of normal brains, and it is not altered in the remaining motor neurons of patients with amyotrophic lateral sclerosis (ALS). NAIP is also located in human fetal neurons and in adult choroid plexus cells. These results suggest that the anti apoptotic molecule NAIP may be important in motor neurons, but it specifically does not appear to be altered in ALS. PMID- 10683822 TI - MRI T2 shortening ('black T2') in multiple sclerosis: frequency, location, and clinical correlation. AB - Abnormal iron deposition occurs in the brains of patients with multiple sclerosis (MS) and may cause MRI T2 shortening ('black T2'; BT2). The frequency, distribution and clinical significance of BT2 in MS is unknown. Analysis of brain MRI scans of 114 MS patients showed BT2 in thalamus (n = 65; 57%), putamen (n = 48; 42%), caudate (n = 27; 24%) and Rolandic cortex (n = 9; 8%). BT2 was significantly related to longer disease duration and advancing neurological disability. Wheelchair-bound patients had worse BT2 in thalamus (p < 0.05), putamen (p < 0.001) and Rolandic cortex (p < 0.05). Patients with secondary progressive disease (n = 34) had worse BT2 in thalamus, putamen and caudate (all p < 0.05) than those with relapsing remitting disease (n = 80). BT2 is proposed as a clinically relevant finding relating to neuronal degeneration in MS. PMID- 10683823 TI - Noradrenaline does not change the mode of discharge of auditory cortex neurons. AB - The mode of discharge of auditory cortex cells was studied during iontophoretic application of noradrenaline (NA). Only seven of 190 cells showed changes in interspike interval distribution during NA application. A similar conclusion was drawn when the analysis focused on 68 cells classified as bursting (n = 15), regular spiking (n = 49) or thin spike (n = 4) cells. Only two bursting cells showed changes in their ISI distribution. The effects on the mode of discharge were independent of the effect on the spike rate and were not a function of cortical depth. These results suggest that the changes in firing mode previously described in vitro occur for a limited percentage of cells and/or for cell types not very often recorded in vivo. PMID- 10683824 TI - Distribution of PDE4A and G(o) alpha immunoreactivity in the accessory olfactory system of the mouse. AB - Distribution of the cAMP-specific phosphodiesterase PDE4A was examined in the accessory olfactory system by immunohistochemistry. Adjacent sections through the vomeronasal organ (VNO) and accessory olfactory bulb (AOB) were alternately immunostained with antibodies against PDE4A or the G-protein alpha subunit G(o) alpha, which labels basal VNO neurons, in order to determine whether PDE4A occurs preferentially in one of two segregated VNO pathways. We found that PDE4A strongly labeled apical VNO neurons and rostral AOB glomeruli. There was virtually no overlap in G(o) alpha and PDE4A staining, and there were no regions of the VNO neuroepithelium or AOB glomeruli not labeled by either antibody. These results identify a potential member of the pheromone transduction cascade in apical neurons, and provide further evidence that the VNO consists of functionally distinct pathways. PMID- 10683825 TI - Activity-related changes in intracellular pH in rat thalamic relay neurons. AB - Activity-related shifts in intracellular pH (pHi) can exert potent neuromodulatory actions. Different states of neuronal activity of thalamocortical neurons were found to differentially modulate pHi. Tonic activity evoked by injection of depolarizing current led to a reversible rise in [H+]i which was nearly abolished in the presence of TTX. Block of voltage-gated calcium channels with I mM Ni2+ reduced the [H+]i transients related to tonic activity. Rhythmic activation of burst discharges caused changes of [H+]i which were decreased by TTX, whereas I mM Ni2+ almost abolished the [H+]i transients. The present results show that different forms of neuronal activity can lead to intracellular acidification caused by different mechanisms, i.e. Na+ and Ca2+ influx through sodium and Ca2+ channels, respectively, and the subsequent activation of a Ca2+/H+ pump. The resulting acidosis is suggested to reduce further Ca2+ influx and prevent excessive neuronal excitation. PMID- 10683826 TI - Suppression of gamma activity in the human medial temporal lobe by sevoflurane anesthesia. AB - We have reported the presence of continuous gamma (30-150 Hz) activity in the human medial temporal lobe (MTL). Since the MTL is involved in learning and memory, we speculated that MTL gamma activity is related to such higher brain functions. It is thus of interest to learn how this activity changes during different states of consciousness. In this study, we recorded electrocorticographic (ECoG) activity directly from the surface of the MTL after various doses of sevoflurane anesthesia. Five epileptic patients underwent electrode placement operations in which electrodes were attached to the surfaces of the MTL and the basal temporal lobe (BTL). Immediately following the operation ECoG was recorded from each patient under four concentrations of sevoflurane anesthesia (1.5, 2.0, 2.5 and 3.0%). Fast Fourier Transform (FFT) analysis was performed on the MTL ECoGs. Under the lowest sevoflurane concentration, MTL gamma activity was observed in all patients. However, gamma activity was progressively suppressed by increased concentrations of sevoflurane, in a dose-dependent manner. Sevoflurane has been known to reduce neuronal excitability in the rat hippocampus in vitro, probably by changing GABAergic inhibition. The reduction of MTL gamma in the present study may be the result of such a mechanism. Although memory function was not tested in this study, the amount of MTL gamma activity may be related to residual memory function during anesthesia. PMID- 10683827 TI - Modulating emotional responses: effects of a neocortical network on the limbic system. AB - Humans share with animals a primitive neural system for processing emotions such as fear and anger. Unlike other animals, humans have the unique ability to control and modulate instinctive emotional reactions through intellectual processes such as reasoning, rationalizing, and labeling our experiences. This study used functional MRI to identify the neural networks underlying this ability. Subjects either matched the affect of one of two faces to that of a simultaneously presented target face (a perceptual task) or identified the affect of a target face by choosing one of two simultaneously presented linguistic labels (an intellectual task). Matching angry or frightened expressions was associated with increased regional cerebral blood flow (rCBF) in the left and right amygdala, the brain's primary fear centers. Labeling these same expressions was associated with a diminished rCBF response in the amygdalae. This decrease correlated with a simultaneous increase in rCBF in the right prefrontal cortex, a neocortical region implicated in regulating emotional responses. These results provide evidence for a network in which higher regions attenuate emotional responses at the most fundamental levels in the brain and suggest a neural basis for modulating emotional experience through interpretation and labeling. PMID- 10683828 TI - Knock down of spinal NMDA receptors reduces NMDA and formalin evoked behaviors in rat. AB - Chronic pain remains a major health problem afflicting an estimated 70% of patients with advanced cancer and inflammatory disorders, and up to 94% of patients with spinal cord injuries. Although progress has been made in the pharmacotherapy of chronic pain management, such as usage of adjuvant drugs and more effective methods of drug delivery, the mainstay of clinical pain management still depends on opiates. NMDA receptor activation, at the level of the spinal cord has been shown to play an important role in the facilitation of nociception (pain) in several animal models. Unfortunately, potent NMDA receptor antagonists, such as MK-801 and APV, have toxic properties and low safety margins that preclude their clinical use. We present evidence which indicates that the use of antisense oligonucleotides targeted to the NMDA-R1 receptor subunit (AS-NMDA-R1), but not sense, abolishes NMDA and formalin induced behaviors. Moreover, we demonstrate that spinal administration of AS-NMDA-R1 results in the abolition of staining for immunoreactive NMDA-R1 in the spinal cord. These data provide novel evidence supporting the feasibility of the use of gene therapy approaches in the management of neuropathic pain. PMID- 10683829 TI - Pro-apoptotic effects of tau mutations in chromosome 17 frontotemporal dementia and parkinsonism. AB - It was recently discovered that mutations of tau cause hereditary frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). Here we report that cultured SH-SY5Y human neuroblastoma cells transfected with mutated tau genes are more vulnerable to apoptotic stimulus. Two kinds of mutations of tau causing FTDP 17 were examined in the present study: one was in exon 10 (N279K) and the other was in exon 12 (V337M). SH-SY5Y cells transfected with either mutated tau were more vulnerable to serum withdrawal, whereas cells transfected with the wild-type tau or vector alone showed no significant change in apoptotic vulnerability. The increase in the intracellular calcium concentration by the serum withdrawal was significantly greater in the SH-SY5Y cells transfected with mutated tau genes than in cells transfected with the wild-type tau or vector alone. These results suggest that mutations of tau might cause FTDP-17 by these pro-apoptotic functions by disrupting the intracellular calcium homeostasis. PMID- 10683830 TI - Evidence that the mismatch negativity system works on the basis of objects. AB - The purpose of this study was to determine whether the system that underlies the mismatch negativity (MMN) of event-related potentials would operate on the basis of objects if stimuli were delivered in such a way as to create the impression of two objects. To this end, tones were alternated between ears with one combination of features for each ear. Deviant tones, which differed from the standard tones of both ears, were delivered separately to each ear. The deviants elicited MMNs only with respect to the standards of the ear to which they were delivered. The data indicate that the MMN system operated on the basis of objects and that the integration of objects occurs preattentively in the auditory system. PMID- 10683831 TI - Nerve growth factor treatment alters Ca2+ pump levels in PC12 cells. AB - Nerve growth factor (NGF) treatment converts rapidly dividing PC12 cells into a neuronal phenotype. To understand the Ca2+ sequestration mechanisms accompanying this differentiation, we examined the endoplasmic reticulum Ca2+ (SERCA) pump levels using two different assays: ATP-dependent azide insensitive oxalate stimulated 45Ca2+ uptake by PC12 cells permeabilized with saponin, and Western blots using a monoclonal antibody which reacts with all the SERCA isoforms. We also examined the reaction to an antibody against the plasma membrane Ca2+ (PMCA) pump. NGF treatment decreased the SERCA pump expression but it increased the PMCA pump level. These results are consistent with a greater role of PMCA pumps in neuronal cells than in most other cells and with an increased role of SERCA pumps during cell proliferation. PMID- 10683832 TI - The N170 occipito-temporal component is delayed and enhanced to inverted faces but not to inverted objects: an electrophysiological account of face-specific processes in the human brain. AB - Behavioral studies have shown that picture-plane inversion impacts face and object recognition differently, thereby suggesting face-specific processing mechanisms in the human brain. Here we used event-related potentials to investigate the time course of this behavioral inversion effect in both faces and novel objects. ERPs were recorded for 14 subjects presented with upright and inverted visual categories, including human faces and novel objects (Greebles). A N170 was obtained for all categories of stimuli, including Greebles. However, only inverted faces delayed and enhanced N170 (bilaterally). These observations indicate that the N170 is not specific to faces, as has been previously claimed. In addition, the amplitude difference between faces and objects does not reflect face-specific mechanisms since it can be smaller than between non-face object categories. There do exist some early differences in the time-course of categorization for faces and non-faces across inversion. This may be attributed either to stimulus category per se (e.g. face-specific mechanisms) or to differences in the level of expertise between these categories. PMID- 10683833 TI - Topiramate depresses carbachol-induced plateau potentials in subicular bursting cells. AB - Intracellular recordings were made in an in vitro slice preparation to establish whether the antiepileptic drug topiramate reduces the depolarizing plateau potentials (PPs) induced in the rat subiculum by intracellular pulses of depolarizing current, in the presence of the cholinergic agonist carbachol (CCh, 70-100 microM). PPs lasted up to about 2 s, and disappeared during application of the muscarinic receptor antagonist atropine. Topiramate (10-100 microM, n = 22 neurons) decreased and eventually abolished in a dose-dependent manner these PPs, even when the function of excitatory amino acid and GABAA receptors was blocked. Hence, topiramate depresses muscarinic receptor-dependent PPs in the rat subiculum, thus suggesting that this form of excitation may represent a target for the mechanism of action of this antiepileptic compound. PMID- 10683834 TI - Developmental prosopagnosia with normal configural processing. AB - The configural processing hypothesis proposes that prosopagnosia results from a domain-general impairment in configural processing, and so predicted that all prosopagnosics would have impaired configural processing. In order to test this prediction, tests of face recognition and configural processing were presented to a developmental prosopagnosic. He was severely impaired in face recognition, but his normal performance on three tests of configural processing disconfirmed the configural processing hypothesis. Additional tests of low-level vision and object recognition found no evidence of impairments with material other than faces. The pattern of spared and impaired face recognition indicates that this case of developmental prosopagnosia is caused by a domain-specific inability to match novel views of faces with previously derived representations. PMID- 10683835 TI - SNAP-25 is present on the Golgi apparatus of retinal neurons. AB - SNAP-25 is a neuronal SNARE protein required for synaptic vesicle exocytosis and neurite outgrowth. Here we show that in addition to synaptic staining, SNAP-25 immunoreactivity is also localized to an intracellular, perinuclear compartment of retinal neurons. Double-labeling with an antibody against the 58 kD resident protein of the trans-golgi network indicates that the intracellular SNAP-25 is localized to the Golgi complex. Immuno-electron microscopic localization of SNAP 25 confirmed its presence on the Golgi apparatus of photoreceptors, bipolar cells, amacrine cells and ganglion cells in the retina. These data implicate SNAP 25 in the trafficking of Golgi-derived vesicles in neurons in addition to the synaptic vesicle cycle. PMID- 10683836 TI - Astrocytes in chronic active multiple sclerosis plaques express MHC class II molecules. AB - To initiate the inflammatory cascade leading to demyelination in multiple sclerosis (MS) T cells have to recognize their specific myelin antigen, which needs to be presented in the context of major histocompatibility (MHC) class II molecules expressed on antigen presenting cells. Whether astrocytes can express MHC class II molecules in vivo is a controversial issue. We performed double labeling immunohistochemistry in postmortem samples from nine patients with MS, three patients with a cerebral infarction and six controls. Astrocytes in controls, in normal appearing white matter in MS, and at the boundary of infarctions were MHC class II negative. In contrast, a subset of astrocytes in active chronic plaques immunostained for MHC class II, indicating potential antigen presenting interactions of astrocytes in MS. PMID- 10683837 TI - A sparse projection from the suprachiasmatic nucleus to the sleep active ventrolateral preoptic area in the rat. AB - The circadian clock of the suprachiasmatic nucleus (SCN) may control the sleep wake cycle by modulating the activity of brain regions important in sleep onset and maintenance, such as the ventrolateral preoptic area (VLPO). The aim of this study was to determine whether the VLPO receives direct projections from the SCN. The retrograde tracer cholera toxin (beta subunit; CT beta) was injected into the VLPO of male rats and the SCN was examined for the presence of labeled, VLPO projecting neurons. After injections restricted to the VLPO only a few labeled cells were found within the SCN, with more labeled cells located around the nucleus. Therefore, the circadian regulation of the VLPO is likely to be achieved through multisynaptic pathways or via a diffusible signal, rather than by direct axonal outputs from the SCN to the VLPO. PMID- 10683838 TI - Parvalbumin-immunoreactive neurons in the human anteroventral thalamic nucleus. AB - We immunohistochemically characterised the expression of the calcium-binding protein parvalbumin in the normal human anteroventral thalamic nucleus (AVN). Two morphologically distinct neuronal populations were found to be parvalbumin immunoreactive (PV-IR): a large population of lightly staining PV-IR neurons and a smaller population of intensely PV-IR neurons. This second type of neuron, which displayed many characteristics normally associated with GABAergic interneurons, has not previously been described in human thalamus. Thus, presumptive thalamic interneurons in the human brain can be further subtyped on the basis of immunoreactivity to parvalbumin. This may have implications for the understanding of thalamocortical function in the normal state and in dysfunctional conditions such as Wernicke-Korsakoff syndrome and schizophrenia. PMID- 10683839 TI - Hypocretin I in the lateral hypothalamus activates key feeding-regulatory brain sites. AB - Hypocretin I (also referred to as orexin A) administered into the lateral hypothalamus (LH) stimulates feeding in rats. We undertook the present study to determine the brain regions activated by LH administration of hypocretin I. Hypocretin I administered into the LH significantly elevated cFos immunoreactivity in the lateral septal area, the central nucleus of the amygdala, the shell of the nucleus accumbens, the bed nucleus of the stria terminalis, the LH, the posterior and dorsomedial hypothalamus, the perifornical, arcuate and paraventricular nuclei and the nucleus of the solitary tract. These data indicate that LH hypocretin I communicates with other key energy regulatory sites within the hypothalamus, the limbic region and the hindbrain, and suggest that these areas are important in the feeding-stimulatory actions of hypocretin I. PMID- 10683840 TI - New aspects of motion perception: selective neural encoding of apparent human movements. AB - Perception of apparent motion operates somewhat differently for objects and human figures. Depending on the interstimulus interval, the latter d may give rise to either perception of a direct path (i.e. biologically impossible) or indirect path (i.e. biologically possible). Here, PET was used to investigate whether a change in brain activity accompanies this perceptual shift. We found neural encoding of apparent motion to be a function of the intrinsic properties of the stimulus presented (object vs human) as well as the kind of human movement path perceived (biomechanically possible vs impossible). Motor and parietal cortex were only involved for possible motion which suggests that these regions are selectively activated to process actions which conform to the capabilities of the observer. PMID- 10683841 TI - Effect of 2-mercaptoacetate and 2-deoxy-D-glucose administration on the expression of NPY, AGRP, POMC, MCH and hypocretin/orexin in the rat hypothalamus. AB - Using in situ hybridization, the mRNA levels encoding neuropeptide Y (NPY), agouti gene-related protein (AGRP), proopiomelanocortin (POMC), melanin concentrating hormone (MCH) and hypocretin/orexin (HC/ORX) were investigated in the rat arcuate nucleus (Arc) and lateral hypothalamic area (LHA) 2 h after a single dose of the glucose antimetabolite 2-deoxy-D-glucose (2-DG; 600 mg/kg) or of the fatty acid oxidation inhibitor mercaptoacetate (MA; 600 mumol/kg). Two hours after 2-DG or MA injection food intake was significantly increased. NPY and AGRP mRNA levels in the Arc were increased by 2-DG but not affected by MA, and MCH mRNA levels in the LHA were increased by both antimetabolites. These results suggest that Arc neurons expressing NPY and AGRP are regulated by changes in glucose, but not fatty acid availability, whereas both factors affect MCH neurons in the LHA. PMID- 10683842 TI - Brain representation of habituation to repeated complex visual stimulation studied with PET. AB - To investigate CNS habituation (i.e. response decrement due to stimulus repetition) the present study used positron emission tomography (PET) to measure regional cerebral blood flow (rCBF) in eight healthy women during two repetitions of complex visual stimuli. Repeated visual stimulation resulted in neural habituation bilaterally in the secondary visual cortex and in the right medial temporal cortex including the amygdala and the hippocampus. Regional CBF in the left thalamus was elevated as a function of repeated stimuli presentations. Thus, repeated presentation of complex visual stimuli result in rCBF habituation in later stages of the visual processing chain. The elevated neural activity in the thalamus might be associated with interruption of further neural transmission related to suppression of non-meaningful behavior. PMID- 10683843 TI - A laboratory study of sleep in Asperger's syndrome. AB - Asperger's syndrome (AS) is a pervasive developmental disorder that may fall along the autistic spectrum. We compared the sleep of eight patients with AS with that of participants matched for age and gender. Patients with AS showed decreased sleep time in the first two-thirds of the night, increased number of shifts into REM sleep from a waking epoch, and all but one patient showed signs of REM sleep disruption. EEG sleep spindles were significantly decreased while K complexes and REM sleep rapid eye movements were normal. Three patients with AS, but none of the comparison participants, showed a pathological index of periodic leg movements in sleep. These observations show that sleep disorders are associated with AS and suggest that defective sleep control systems may be associated with the clinical picture of AS. PMID- 10683844 TI - Subdural applications of NO scavenger or NO blocker to the cerebellum depress the adaptation of monkey post-saccadic smooth pursuit eye movements. AB - We examined pharmacologically whether cerebellar long-term depression (LTD) may play a role in the adaptation of smooth pursuit eye movements in two Macaca fuscata, which were trained to pursue a target moving in the horizontal plane in a 3 degrees step-10 deg/s ramp mode. The monkeys showed small catch-up saccades followed by 6-8 deg/s post-saccadic pursuit movements. Adaptation of the post saccadic pursuit velocity was induced by repetition of acceleration of the target to 20 deg/s after the catch-up saccades. Injections of 0.1 mM hemoglobin or 20 mM NG-monomethyl-L-arginine solution into the subdural space above the paraflocculus flocculus scarcely affected the post-saccadic pursuit velocity, but markedly depressed its adaptation. These observations suggest that cerebellar LTD may underlie the adaptation of smooth pursuit. PMID- 10683845 TI - Dorsolateral prefrontal cortex and the implicit association of concepts and attributes. AB - The Implicit Association Test (IAT) examines the differential association of two object categories (e.g. flower and insect) with attribute categories (e.g. pleasant and unpleasant). When items from congruent categories (e.g. flower + pleasant) share a response key, performance is faster and more accurate than when items from incongruent categories (e.g. insect + pleasant) share a key. Performing incongruent word classification engages inhibitory processes to overcome the prepotent tendency to map emotionally congruent items to the same response key. Using fMRI on subjects undergoing the IAT, we show that the left dorsolateral prefrontal cortex, and to a lesser extent the anterior cingulate cortex, mediate inhibitory processes where manipulation of word association is required. PMID- 10683846 TI - Branchiomotor activities in mouse embryo. AB - Using a novel isolated hindbrain in vitro preparation, we demonstrate that, in the mouse, branchiomotor activities from trigeminal, facial, glossopharyngeal and vagal nerves start during segmentation, a crucial and conserved period of hindbrain embryogenesis. At embryonic day (E) 10.5, branchiomotor nerves are independently active in bursts, become coactive at a low frequency (about 0.5 min 1) at E12.5, before high frequency (about 15 min-1) fetal breathing starts at E14.5. Comparison with observations in chick reveals a transient episodic rhythmic pattern highly similar in mouse at E13.5 and chick at E7. This pattern is proposed as a marker identifying a phylotypic stage during the development of hindbrain neuronal networks in vertebrates. PMID- 10683847 TI - The representation of the plegic hand in the motor cortex: a combined fMRI and TMS study. AB - TMS mapping and fMRI were used to investigate changes in the motor cortex representation of the hand in a patient with complete loss of right hand function following traumatic avulsion of the cervical roots C7 and C8. Both TMS and fMRI demonstrated an expansion of the motor representation of the forearm into the hand area contralateral to the injured side. fMRI of the hand area, however, revealed that this area could still be activated when the patient was instructed to imagine finger tapping with his plegic hand. These results indicate that the plegic hand is still represented in the motor cortex, despite the fact that the same cortical area is also now active during movements involving forearm muscles. PMID- 10683848 TI - Alteration of veratridine neurotoxicity in sympathetic neurons during development in vitro. AB - Neurotoxic effects of veratridine, the activator of voltage-dependent Na+ channels, were examined at various stages of in vitro development of superior cervical ganglion cells dissociated from newborn rats. Veratridine neurotoxicity did not occur in 1DIV (days in vitro) neurons, but occurred in 7DIV neurons, both of which depend on NGF for survival, but elevated K+ supports only the latter. TUNEL and electron microscopic analyses revealed that 7DIV neurons underwent both apoptotic and necrotic cell death. Veratridine was also toxic to 21DIV neurons which are independent of NGF for survival. Nuclear features of apoptosis, however, were greatly reduced in these neurons undergoing cell death, suggesting that nuclear vulnerability is also subject to developmental regulation in vitro. PMID- 10683849 TI - Mood state and brain electric activity in ecstasy users. AB - Resting EEG during open and closed eyes and subsequent mood ratings were obtained from 15 Ecstasy users and 14 Ecstasy-naive controls. Absolute spectral power on the scalp, and the three-dimensional, intracerebral distribution of neuroelectric activity using low resolution brain electromagnetic tomography (LORETA) were computed. LORETA revealed global increases of theta, alpha 1 and beta 2/3 power during eyes open in Ecstasy users, and spectral analyses revealed a right posterior increase of alpha 2 power (confirmed by LORETA) and increased beta band activity during open eyes. Ecstasy users had higher levels of state depressiveness, emotional excitability and a trend-level increase in state anxiety. The observed differences may be related to regular exposure to Ecstasy or other illicit drugs, or may be pre-existing. PMID- 10683850 TI - Histamine H3-receptor activation inhibits dopamine synthesis in rat striatum. AB - Unilateral 6-hydroxydopamine lesion to rat substantia nigra pars compacta resulted in a modest, but significant, decrease in the specific binding of N alpha-[methyl-3H]histamine (19 +/- 5% reduction) to synaptosomal membranes from ipsilateral striata. Dopamine synthesis was assessed in striatal slices by determining [3H]DOPA accumulation after inhibition of DOPA decarboxylase. [3H]DOPA synthesis induced by 50 mM K+ (151 +/- 4% of basal) was prevented by either Ca2+ removal or by Ni2+. Depolarization-stimulated [3H]DOPA accumulation was reduced by the selective H3-agonist immepip (100 nM; 68 +/- 7% inhibition). The effect of immepip was reversed by thioperamide (100 nM), a selective H3 antagonist. Taken together, our results indicate that histamine modulates striatal dopamine synthesis by acting at H3-receptors located on dopaminergic nerve terminals. PMID- 10683851 TI - Selective increase in cellular A beta 42 is related to apoptosis but not necrosis. AB - Amyloid beta protein ending at 42 (A beta 42) plays an important role in the pathology of Alzheimer's disease (AD). Here we show an increase in cellular A beta 42 in damaged neurons, with both ELISA and immunocytochemistry. The cellular A beta 42 increase was caused by 3-day treatments with H2O2, etoposide or melphalan, all of which induce genotoxic apoptosis, but not by treatment with sodium azide, which causes necrosis. Secreted A beta was similarly decreased with all these treatments. The cellular A beta 42 increase appeared even with minimal damage (ELISA) and A beta 42-positive cells were TUNEL negative (double staining), indicating that any early apoptosis mechanism may induce the cellular A beta 42 increase. Thus, neuronal apoptosis and cellular A beta 42 increase may be linked in a way that contributes importantly to AD pathology. PMID- 10683852 TI - Effects of dizocilpine (MK 801) on noradrenaline, serotonin and dopamine release and uptake. AB - In the present study, we examined the actions of the NMDA antagonist dizocilpine (MK801) on electrically evoked release and uptake of noradrenaline (NA) in the locus coeruleus (LC), serotonin (5-HT) in the dorsal raphe nucleus (DRN) and dopamine (DA) in the nucleus accumbens (NAc), measured by fast cyclic voltammetry (FCV) in rat brain slices. Dizocilpine (10 microM) significantly increased NA (to 248 +/- 15%) and 5-HT release (to 184 +/- 29%) and slowed monoamine uptake in the LC (t1/2 = 853 +/- 129%) and the DRN (t1/2 = 387 +/- 70%), respectively. However, dizocilpine had no effect on DA release or uptake in NAc. Actions on monoamines are thus likely and should be considered in the interpretation of data regarding dizocilpine. PMID- 10683853 TI - Early damage of sympathetic neurons after co-culture with macrophages: a model of neuronal injury in vitro. AB - Since activated immune cells may damage peripheral nerves during inflammation, we developed a co-culture model that permits the direct study of macrophage-induced neuronal damage. Sympathetic neurons were enzymatically isolated from neonatal mice and co-cultured with increasing numbers of peritoneal macrophages for 24 h. This caused rapid neuronal cell death, reducing neuronal number by 24.1 +/- 4% with the addition of 11.5 x 10(3) macrophages, representing a ratio of 8 macrophages per neuron. Nuclear analysis showed that cell death occurred by both apoptosis and necrosis. These effects were not mimicked by addition of macrophage conditioned medium, and were prevented by 10 microM dexamethasone. Although no appreciable neuronal death occurred beyond 24 h, the density of neurites was decreased between 1 and 2 days of co-culture (p < 0.05). There is, therefore, a rapid induction of cytotoxicity by macrophages after their addition to the neuronal cultures, followed by axonal damage without neuronal cell death. PMID- 10683854 TI - Effects of dalargin on excitation induced by L-glutamate agonists in the frog vestibular organs. AB - We have used an electrophysiological approach to investigate the action of a synthetic analog of leu-enkephalin dalargin (DAL) on chemically induced afferent activity in the frog vestibular organs. Administration of 5.0 microM kainic acid (KA), 5.0 microM (AMPA) and 50 microM NMDA produced an increase in the frequency of the resting discharge. Firing evoked by KA, AMPA or NMDA could be depressed by administration of 1 nM Dal by 55.5 +/- 9.9% (n = 10, p < 0.05), 64.5 +/- 11.2% (n = 13, p < 0.05) and 21.3 +/- 11.1% (n = 14, p = 0.051), respectively. Thus, the frequency decrease under NMDA was statistically non-significant. These results show that non-NMDA, but not NMDA subtypes of receptors are mostly involved in opioid action at the vestibular organs of the frog. PMID- 10683855 TI - Somatosensory areas in man activated by moving stimuli: cytoarchitectonic mapping and PET. AB - This study was performed to identify neuronal populations in the somatosensory areas engaged in discrimination of moving stimuli on the skin. Changes in regional cerebral blood flow (rCBF) were measured with positron emission tomography (PET) and correlated with cytoarchitectonic sensorimotor areas 4a, 4p, 3a, 3b, and 1. Volunteers discriminated differences in the speed of a rotating brush stimulating the palmar surface. Discrimination of moving stimuli, contrasted to rest, increased the rCBF mainly in primary somatosensory (SI) area 1, but also in area 3b. The parietal operculum (PO) was activated bilaterally. We conclude that area 1 is the area in SI which is mainly responding to discrimination of moving stimuli and that the PO contains several regions engaged in the discrimination of fast transient stimuli. PMID- 10683856 TI - Role of central adenosine in the respiratory and thermoregulatory responses to hypoxia. AB - No reports are available about the role of central adenosine in the respiratory and thermoregulatory responses to hypoxia in conscious rats. We therefore measured ventilation (VE) and body temperature (Tb) before and after intracerebroventricular injection of saline or aminophylline (adenosine antagonist), followed by a 30-min period of hypoxia exposure. Aminophylline did not change VE or Tb during normoxia; however, during hypoxia, it caused a significant increase in VE, and significantly attenuated hypoxic hypothermia. The present data indicate that central adenosine has an inhibitory effect on hypoxic hyperventilation and partially causes hypoxic hypothermia, suggesting that the ventilatory and metabolic interaction during hypoxia does not involve opposing mechanisms. PMID- 10683857 TI - Localization of the tetrodotoxin-resistant sodium channel NaN in nociceptors. AB - Tetrodotoxin-resistant sodium currents contribute to the somal and axonal sodium currents of small diameter primary sensory neurons, many of which are nociceptive. NaN is a recently described tetrodotoxin-resistant sodium channel expressed preferentially in IB4-labeled dorsal root ganglion (DRG) neurons. We employed an antibody raised to a NaN specific peptide to show that NaN is preferentially localized along axons of IB4-positive unmyelinated fibers in the sciatic nerve and in axon terminals in the cornea. NaN immunoreactivity was also found at some nodes of Ranvier of thinly myelinated axons of the sciatic nerve, where it was juxtaposed to Kv1.2 potassium channel immunoreactivity. This distribution of NaN is consistent with a role for NaN sodium channels in nociceptive transmission. PMID- 10683858 TI - Estimation of photoparoxysmal response elicited by half-field visual stimulation. AB - We examined photoparoxysmal responses (PPRs) elicited by half-field visual stimulation with deep-red flicker light to determine the neurophysiological features of photosensitive epilepsy (PSE). EEG revealed two types of PPRs. One had the focal spike in the occipital region and the other in the temporo occipital region at the contralateral hemisphere. The equivalent current dipoles of these types were located at the occipital cortex and the inferior temporal (IT) cortex, respectively. These cortices comprise one of the main pathways in the visual system, and they play important roles in color discrimination. Thus, we propose that the visual system, especially the occipital cortex and the IT cortex, might be involved in the generator mechanism of PSE. PMID- 10683859 TI - Sound-induced laryngeal and respiratory reflexes originate from vestibular afferents. AB - To determine whether the auditory or vestibular system causes the sound-induced laryngeal reflex, which has been considered to participate in the auditory feedback control of vocalization, click-induced laryngeal responses were compared before and after sectioning of the cochlear and/or vestibular nerves in cats. The sound-induced reflex modulation of respiratory muscle activity was also investigated, because respiratory movement is important for vocal control. Sectioning of the cochlear nerves had little influence on these responses. In contrast, sectioning of the vestibular nerves abolished these responses. It was concluded that the sound-induced laryngeal and respiratory reflexes are attributed to the vestibular system. PMID- 10683860 TI - MPTP induces alpha-synuclein aggregation in the substantia nigra of baboons. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity reproduces many of the features of Parkinson's disease (PD). alpha-Synuclein has been identified as a prominent component of the Lewy body (LB), the pathological hallmark of PD. MPTP-treated primates have been reported to develop intraneuronal inclusions but not true Lewy bodies. We administered MPTP to baboons and used a monoclonal alpha synuclein antibody to define the relationship between neuronal degeneration and alpha-synuclein immunoreactivity in the substantia nigra. MPTP-induced neuronal degeneration was associated with the redistribution of alpha-synuclein from its normal synaptic location to aggregates in degenerating neuronal cell bodies. alpha-Synuclein aggregation induced by MPTP models the early stages of Lewy body formation and may be a fundamental step in the evolution of neuronal degeneration in PD. PMID- 10683861 TI - Serotonin transporter gene polymorphism and antidepressant response. AB - We examined allelic polymorphisms of the serotonin transporter (5-HTT) gene and antidepressant response to 6 weeks' treatment with the selective serotonin reuptake inhibitor (SSRI) drugs fluoxetine or paroxetine. We genotyped 120 patients and 252 normal controls, using polymerase chain reaction of genomic DNA with primers flanking the second intron and promoter regions of the 5-HTT gene. Diagnosis of depression was not associated with 5-HTT polymorphisms. Patients homozygous l/l in intron 2 or homozygous s/s in the promoter region showed better responses than all others (p < 0.0001, p = 0.0074, respectively). Lack of the l/l allele form in intron 2 most powerfully predicted non-response (83.3%). Response to SSRI drugs is related to allelic variation in the 5-HTT gene in depressed Korean patients. PMID- 10683862 TI - Role of dopamine D3 receptors in thermoregulation: a reappraisal. AB - Dopamine agonist-induced hypothermia has been proposed to be mediated by the D3 receptor (D3R), as it is elicited by (+)7-OH-DPAT and antagonized by S 14297, two putative D3R-preferential ligands. Here we show, however, that S 14297 is a full and partial agonist at D3R and D2R, respectively. Hypothermia was induced in rats by agonists with potencies correlated with their D3R and D2R functional potencies, and was reversed by antagonists, with a rank order of potency typical of the D2R. Moreover, BP 897, a highly potent and selective but partial D3R agonist was inactive in producing hypothermia or reversing (+)7-OH-DPAT-induced hypothermia. (+)7-OH-DPAT was as potent and efficient in inducing hypothermia in wild-type as in D3R-deficient mice. Hence, our results suggest that hypothermia does not result from a selective stimulation of the D3R. PMID- 10683863 TI - Synthesis of new derivatives of 1,2,3,4,7-pentamethylbicyclo[2.2.1]hept-2-ene-5,6 dicarboximide with an expected anxiolytic activity. AB - The preparation of a number of derivatives of 1,2,3,4,7 pentamethylbicyclo[2.2.1]hept-2-ene-5,6-dicarboximide with potential anxiolytic activity has been described. The aim of our study was to obtain new analogues of tandospirone, that is derivatives of cyclic imides [1]. PMID- 10683864 TI - Synthesis of some N-substituted 3,4-pyrroledicarboximides as potential CNS depressive agents. AB - As a continuation of our work on N-[4-aryl(heteroaryl)piperazin-1-ylalkyl]-3,4 pyrro ledicarboximides, which were characterized by strong analgesic activity and CNS depressive action, several novel N-substituted 3,4-pyrroledicarboximides were prepared and eleven representatives were examined in a series of in vivo CNS tests. A few of these compounds displayed a similar profile of biological selectivity to that of 3,4-pyrroledicarboximides described previously; their structure-activity relationships are discussed. PMID- 10683865 TI - [Color reactions for identification of nalidixic acid]. AB - Nalidixic acid (1) gives with 2-naphthol a yellow charge-transfer complex. The 7 methyl group of 1 condenses with vanillin (2) and Ehrlich's reagent (4) to the coloured (E)-benzylidene compounds 3 and 5. Treating 1 with thionyl chloride and subsequent reaction with aminopyrazolone (6) and sodium acetate leads to a mixture of trichloronalidixic acid (7) and its 3-carboxamide 8. The trichloromethyl group of 7 is converted with 6 in pyridine to form the amide 9. Nalidixic acid reacts with 1,3-dimethylbarbituric acid (10) in acetanhydride/acetic acid to yield the polymethine dyes 11-13, whose structures are confirmed by X-ray crystal structure analysis. The dyes 3 and 12 inhibit the growth of staphylococcus aureus and Escherichia coli, respectively. PMID- 10683866 TI - [Synthesis of 4,5-dihydro-1,2,4-oxadiazoles from N-unsubstituted amidoximes]. AB - 4,5-Dihydro 1,2,4-oxadiazoles can be synthesized from aromatic and araliphatic amidoximes by cyclocondensation with aldehydes and ketones. Resulting heterocycles differ in substitution at C-3 and C-5 showing the scope of the simple reaction. PMID- 10683867 TI - Synthesis and anticonvulsant activity of some amino acid derivatives. Part 3: Derivatives of Ala, Arg, Tzl, Gly and chi Abu. AB - Ten amino acid derivatives as antagonists of the excitatory amino acid (EAA) receptor and anticonvulsant activity have been designed. Five of these compounds supposed to show rather strong and five of them rather weak action as it was expected on the base of their hydrophobicity. All the compounds were synthesized and then evaluated in mice in the maximal electroshock seizure (MES) test, the subcutaneous Metrasol seizure threshold (scMet) test and the rotorod neurotoxicity (Tox) test. Four of the obtained compounds have shown high activity (three received class I and one class II) and six were classified in class III according to the classification of the Anticonvulsant Screening Project (ASP) of the Antiepileptic Drug Development Program (ADDP) of NINDS. One of the compounds classified in class I (10) was tested quantitatively following i.p. administration in mice. It has a MES ED50 = 29.05 b.w. and protective index (PI) of 3.77. PMID- 10683868 TI - Synthesis of new pyrido[4',3':4,5]thieno[2,3-d]-1,2,4-triazolo[3,4-c]pyrimidines and a 5,6-dihydro-1,2,4-triazolo[4",3":1',2']pyrido[4',3':4,5]thieno [2,3-d] pyrimidine ring system. AB - A series of substituted pyrido[4',3':4,5]thieno[2,3-d]-1,2,4-triazolo[3,4 c]pyrimidines 4-6, 8, pyrido[4',3':4,5]thieno[2,3-d]-1,2,4-triazolo[3,4 c]pyrimidines 11-13 and 5,6-dihydro-1,2,4 triazolo[4",3":1',2']pyrido[4',3':4,5]thieno[2,3-d] pyrimidines 16-19 have been synthesized from 3, 10 and 15 through the reaction with orthoesters and carbon disulphide, respectively. PMID- 10683869 TI - [A 1-hydroxyindole-2-carboxylic acid and a 9-hydroxy-beta-carboline-4-carboxylic acid from a nifedipine analog biscyanoethyl ester]. AB - Bis(2-cyanoethyl) 2,6-dimethyl-4-(2-nitrophenyl)-1,4-dihydropyridine-3,5 dicarboxylate (3) reacts with sodium hydroxide solution to yield the 1 hydroxyindole-2-carboxylic acid 7 and the 9-hydroxy-beta-carboline-4-carboxylic acid 13. The structures of 7 and 13 were elucidated by derivatization and by spectroscopic methods. Bis(2-cyanoethyl) 2,6-dimethyl-4-(2-nitrosophenyl)pyridine 3,5-dicarboxylate (22) obtained by irradiation of 3 reacts with sodium hydroxide solution to give the cyclic hydroxamic acid 23 whose structure is proven by an independent synthesis. PMID- 10683870 TI - Fluorescent-labeled ligands for the benzodiazepine receptor. Part 1: Synthesis and characterization of fluorescent-labeled benzodiazepines. AB - Because radioactive labeled ligands in receptor assays have several disadvantages, we synthesized a number of fluorescent-labeled benzodiazepines. Several fluorophores were attached at different positions of 1,4-benzodiazepine molecules in order to assess the impact of the fluorophores and their coupling position on the affinity for the benzodiazepine receptor. Besides the 1,4 benzodiazepines, the 1,2-annelated 1,4-benzodiazepines were also used for labeling. A metabolite of flumazenil (18), desethylflumazenil (Ro15-3890, 19), was labeled with the fluorophore 4-bromomethyl-7-methoxycoumarin, with and without the incorporation of a spacer chain, yielding the methyl-methoxycoumarin (Mmc) derivatives Mmc-Ro15-3890 (20a) and Mmc-O-CO-(CH2)3-Ro15-3890 (20b), respectively. After the synthesis, the fluorescent-labeled benzodiazepines were purified by HPLC, using an analytical RP-C18 column. For the purification of 20b, the chromatographic system was optimized, using multi-criteria decision making (MCDM) techniques. The binding affinities for the benzodiazepine receptor and the fluorescence characteristics were determined for the resulting products. PMID- 10683871 TI - Determination of tetrahydrozoline hydrochloride and fluorometholone in pharmaceutical formulations by HPLC and derivative UV spectrophotometry. AB - Two methods for the quantitative determination of tetrahydrozoline hydrochloride (1) and fluorometholone (2) in pharmaceutical eye drops (Efemoline) are described. The procedures are based on derivative UV spectrophotometry and HPLC. In the former method, d2A/d lambda 2 values were measured in methanol at 226 and 282 nm for 1 and 2, respectively. The relative standard deviations for the method were found to be 1.06% for 1 and 0.98% for 2. The latter method based on a reversed phase HPLC system using a Partisil 5 ODS analytical column. The mobile phase used for the separation of 1, 2 and internal standard (lidocaine) was methanol/acetonitrile/water (50:50:10 v/v) and the compounds in the eye drops were detected at 220 nm using an UV detector. The relative standard deviations for the HPLC method were determined to be 0.61% and 0.50% for 1 and 2, respectively. The proposed methods, which give thoroughly comparable data, are simple, rapid, and allow precise and accurate results and could be used for commercial formulations containing tetrahydrozoline hydrochloride and fluorometholone in combination. PMID- 10683872 TI - ["Non-solvent shock agglomeration technology" as a new alternative method for work on ibuprofen. 4. Production and evaluation of quick release forms]. AB - During compaction of shock-agglomerated S(+)ibuprofen it was of interest if and how far the sometimes strongly differing quality or the origin of the source material has effects on the tabletting properties and on tablet quality. Moreover, conventional and shock agglomerated substances are compared with regard to the parameters mentioned. The technology of "non-solvent shock agglomeration" results in substances suitable for direct tabletting. Additionally, the resulting comprimates have characteristics which can be clearly traced back to the special quality of the ibuprofen-shock agglomerates. By using different agents in the process of substance preparation specific galenic properties can be achieved. PMID- 10683873 TI - Interactions between food components and drugs. Part 8: Effect of pectins and bile acid preparations forming stable mixed micelles on transport of quinine in vitro. AB - Interactions between quinine and acetylated pectin, amidated pectin and pectin with blockwise arrangement of the free carboxyl groups as well as interactions between quinine and bile salt preparations forming stable mixed bicelles have been investigated. A diffusion cell with two compartments and an artificial lipid membrane and a filter-grown colon carcinoma cell line (Caco-2) have been used. Depending on structural parameters, pectin preparations diminished the rate of permeation of the drug. Above the critical micelle concentration, the bile salt preparations influence the quinine transport stronger than the pectin preparations. The strongest inhibition of the quinine permeation showed a stable mixed micelle preparation consisting of glycodeoxycholate, palmitic acid and lecithin. The Caco-2 cell line appears to be not as suitable as artificial lipid membranes to study drug transport in the presence of the bile salt preparations. PMID- 10683874 TI - Synthesis of 1-amino-6,7,8,8a-tetrahydroacenaphthene and its effect on the inhibition of the MAO-enzyme at the brain cortex and liver level. AB - (+/-)-1-Amino-6,7,8,8a-tetrahydroacenaphthene was synthesized and evaluated as a novel drug acting on the dopaminergic system. It was shown that the new compound displays activity as MAO inhibitor. PMID- 10683875 TI - Antitussive activity of a glucuronoxylan from Rudbeckia fulgida compared to the potency of two polysaccharide complexes from the same herb. AB - An alkali-extracted low-molecular glucuronoxylan and two water-extractable polysaccharide complexes isolated from various parts of Rudbeckia fulgida were tested for antitussive activity on mechanically induced cough in nonanaesthetized cats. Glucuronoxylan consisted of a (1-->4)-linked beta-D-xylopyranosyl backbone with about 18% of 4-0-methyl-D-glucuronic acid attached to 0-2 of the chain xylose residues. The polysaccharide complexes differed from each the other regarding the in qualitative and quantitative composition of the sugar components. It was found that peroral administration of all the compounds led to a significant suppression of the cough reflex without negative influence on expectoration. Glucuronoxylan and the complex from the aerial parts of the herb exhibited much higher antitussive activity than the complex from the roots which did not contain any uronic acid component. Their activity (48.2% and 46.5%, respectively) highly surpassed the activity of the complex from the roots (23.5%) as well as that of the peripherally acting drugs dropropizine (28.3%) and prenoxdiazine (24.7%). PMID- 10683876 TI - A common variant of the angiotensinogen gene and the risk of coronary artery disease in a German population. AB - The thymidine to cytosine transition at position 704 in exon 2 of the angiotensinogen gene leads to the amino acid substitution of threonine for methionine (T235 variant) and is responsible for elevated plasma levels of angiotensinogen. To examine the influence of T235 on the risk of coronary artery disease (CAD) we genotyped 184 CAD patients, 77 controls in whom CAD was excluded angiographically, and 155 healthy controls without signs of CAD by polymerase chain amplification and restriction enzyme digestion. Allele frequencies for A (wildtype) and a (mutant allele) in the total study population were 0.538 and 0.462, 0.536 and 0.464 in the healthy controls, and 0.481 and 0.519 in patients with excluded CAD, respectively. The allele frequencies and the genotype distribution in these groups did not show a significant difference. In conclusion, we did not observe an association between the T235 variant of the angiotensinogen gene and the risk of CAD. PMID- 10683877 TI - Cytotoxicity of triterpenoid saponins. Part 2: Relationships between the structures of glycosides of polygalacic acid and their activities against pathogenic Candida species. AB - Glycosides of polygalacic acid (2 beta,3 beta,16 alpha,23-tetrahydroxy-olean-12 ene-28-oic acid) is isolated from the aerial parts of Solidago virgaurea L. subsp. virgaurea, Heteropappus altaicus (Willd.) Novopokr. and Heteropappus biennis (Ldb.) Tamamsch. or produced by degradation of these genuine saponins were tested against humanpathogenic strains of Candida albicans, C. glabrata, C. krusei and C. tropicalis using a micro-dilution assay. The antifungal action can be influenced the variation of the etherglycosidically bonded carbohydrate units at C-3 as well as of the acylglycosidically bonded oligosaccharide at C-28 of the aglycone. PMID- 10683878 TI - Screening of selected plant extracts for in vitro inhibitory activity on HIV-1 reverse transcriptase (HIV-1 RT). AB - Methanolic-aqueous extracts of 70 plants were investigated for their ability to inhibit HIV-1 reverse transcriptase activity in vitro. Two thirds of the extracts screened showed more than 50% inhibition. Two extracts inhibited the enzyme completely while four exhibited more than 90% inhibition. Tannins as nonspecific HIV-1 RT inhibitors were detected and removed from the extracts. The IC50 values of the most potent extracts after the removal of tannins for the HIV-1 RT inhibition are as follows: Sambucus racemosa 0.017 mg/ml and Geranium phaeum 0.067 mg/ml. Daunomycine was chosen as a standard substance in the non radioactive immuno assay used for screening. As a result from the future isolation and characterization of these compounds, new leading structures are expectable. PMID- 10683879 TI - Isolation and structure elucidation of ligustroflavone, a new apigenin triglycoside from the leaves of Ligustrum vulgare L. AB - A new flavone, apigenin-7-O-beta-(2",6"-di-alpha-rhamnopyranosyl)-glucopyranoside , named ligustroflavone, was isolated from the leaves of common privet (Ligustrum vulgare L., Oleaceae), whose popular use was well known in the Mediterranean historical medicine and ethnomedicine as anti-inflammatory. The structures of other five apigenin and luteolin derivates, isolated from the polar fractions of the methanolic leaf extracts, were elucidated. PMID- 10683880 TI - [Aphasia research and speech localization in the brain]. AB - Aphasia research has become an acknowledged branch of modern cognitive neuropsychology research whose aim is to explore more fully the structures of knowledge and of cerebral processes which might both be affected in patients with aphasia. Up to the second half of this century, a model based on a specific cerebral localisation of language processes had emerged based on brain localisation research by Broca and Wernicke (among others). New modern neuroimaging techniques, however, such as computed tomography and magnetic resonance imaging (MRI), but also functional imaging modalities such as positron emission tomography or functional MRI, have modified these concepts. It emerges that in comprehension as well as in production of language, not only a few well defined centres are responsible for the activity, but there is a synchronised activity in large neuronal networks connecting various regions located both in the cortex and in the deep subcortical structures; today, this activity can be demonstrated best in a non-invasive and reproducible way with functional MRI. PMID- 10683881 TI - [High-dose chemotherapy with autologous bone marrow transplantation: 11 years' experience in Zurich]. AB - High-dose chemotherapy with autologous bone marrow or peripheral blood stem cell transplantation has gained widespread acceptance for the treatment of certain malignancies. Since the introduction of this therapy in 1988 we have treated 272 patients. Indications for high-dose chemotherapy were high-risk large cell lymphoma and lymphoblastic or Burkitt lymphoma in first remission (73 patients), non-Hodgkin's lymphoma in chemosensitive relapse (65 patients), Hodgkin's lymphoma in relapse (52 patients), germ cell tumours with inadequate response to chemotherapy (34 patients), multiple myeloma (29 patients), and other malignancies (19 patients). Treatment mortality was 1.8%. The 3-year event-free survival and overall survival for all patients were 48 and 61% respectively. High dose chemotherapy with autologous stem cell transplantation has become a safe procedure and is considered the treatment of choice for relapsed large cell lymphoma, relapsed Hodgkin's disease, stage II or III multiple myeloma, and germ cell tumours with inadequate response to cisplatin-based chemotherapy. In other situations, including aggressive lymphoma with risk factors, acute leucaemia or breast cancer, the superiority of high-dose over conventional chemotherapy remains to be proven. Patients with such diseases should not receive high-dose chemotherapy outside a controlled clinical study. PMID- 10683882 TI - [Postpartum footdrop]. AB - We report on a rare peripartal neuropathy of the leg caused by prolonged difficult labour. Immediately after delivery two patients complained of unilateral footdrop and numbness in the leg. The footdrop was most probably due to compression of the lumbosacral trunk exposed to the foetal head. This trunk contains fibres from lumbar roots L4 and L5 and connects the lumbar with the sacral plexus. The outcome was favourable in both patients. If subsequent pregnancies occur, caesarean section may be indicated. PMID- 10683883 TI - [Spontaneous bacterial peritonitis with Streptococcus constellatus in an HIV positive patient]. AB - Liver diseases are an important cause of high morbidity and mortality in HIV infected patients, and liver cirrhosis is the commonest cause of ascites in this population. We describe the case of a 38-year-old HIV-positive male (CDC stage B3, CD4 cell count 199/mm3) with a history of hepatitis C-associated liver cirrhosis. Following pneumonia he developed spontaneous bacterial peritonitis due to Streptococcus constellatus. Clinically noticeable was the gradually worsening course with few symptoms, despite the initially high ascitic fluid leucocyte count of over 11,000/microliter, but a favourable response to betalactam antibiotics. PMID- 10683884 TI - [Value of radiosynovectomy in rheumatology and orthopedics]. AB - For many years radiosynovectomy has proved its worth in the treatment of inflammatory joint disease. More recently there has also been growing interest in the use of this method for outpatient treatment. In this overview we discuss the established and new indications. With increasing use of the method, interest should be focused on adequate quality control. Accordingly, basic requirements for correct performance of radiosynovectomy in clinical practice are addressed. Our results, based on five years' experience of radiosynovectomy in outpatients, have shown improvement of clinical symptoms in 60-70% of cases. However, success rates of this kind require high quality standards and close cooperation with the referring rheumatologists. PMID- 10683885 TI - [Acute tinnitus]. PMID- 10683886 TI - [Medical discretion and data protection in medicine]. PMID- 10683887 TI - [Medical discretion in the past and today]. AB - It is based on old traditions that a physician and other medical personal have to be diskrete about their conversation with patients as well as about diagnostic and therapeutical measures. The roots are found in the early times when priests were working as medical therapists. Well known are the Asclepiads in old Greece and later on in the Roman Empire. They were priests in the tempels of the god Asclepios and the famous Hippocrates of Cos was also an Asclepiad and nevertheless the founder of western scientific medicine. The historian of medicine can demonstrate a steady development of the professional discretion observed by physicians and surgeons on different political, cultural and scientific conditions until today. PMID- 10683888 TI - [Data protection as a personal right. Government Agency]. AB - The sector-specific secrets are traditionally regarded as the historical predecessors of general data protection law. The lecture demonstrates that they are not only aimed at protecting the individual's right to self-determination of the handling of his personal data. It also indicates that those particular secrets are, at the same time, serving for further purposes which are of importance in their respective sectors. The general (omnibus) data protection statues are given attention in the light of the beginning of automated information processing and in their role of guaranteeing the right to personal privacy, a role which the Federal Constitutional Court underlined in its census decision of 1983 and which became of general relevance in the course of the further developments. In conclusion, the report particularizes that, inter alia, statutory adaptations and amendments are necessary in order to preserve the effects of the doctor-patient confidentiality under the conditions of modern data processing. The examples given are comprising: health data on chip cards, health data in networks and the electronic record of the patient. PMID- 10683889 TI - [Medical discretion towards relatives, colleagues and the media from the physician's point of view]. AB - Three aspects are examined more closely: discretion towards relatives, medical colleagues and the media. Medical discretion has a long tradition and forms an indispensable part of the medical ethos. Physicians, however, often find it difficult to cope with the diversity of legal regulations. Legal regulations manifest themselves in the Social Code with its different books of legislation; in the Federal Data Protection Law; in the Federal Epidemics Law; in the Criminal Code; in the Code of Criminal Procedure. Physician's discretion is an indispensable prerequisite for the confidential relationship between doctor and patient, thus enhancing the success of medical treatment. Consequently, this discretion must never be jeopardized. First of all, it protects the patient's privacy and personal rights. The discretion towards relatives bears different facets which are considered more closely: these include the patient's declaration to unconscious patients, therapeutical decisions. Medical discretion must also be observed among fellow doctors. This is unproblematic if the patient remains informed in the case of joint/further treatment. The following cases are problematic: consulting in additional physicians without the patient's knowledge; joint practices and practice networks. As far as the media are concerned, there is no case whatsoever that would justify a patient being presented in the media without his or her consent. PMID- 10683890 TI - [Medical confidentiality towards family, colleagues and the media from the legal point of view]. AB - The more medical confidentiality is endangered in a society based on the exchange of information especially by the welfare system and a health care system relying on the division of labour, the more it must be protected by the law of professional rules and regulations, civil and criminal law. First of all, medical confidentiality refers to the patient who will only confide in his doctor if he can be sure that his secrets will be kept. The individual patients' interests lead directly to the interests of the public health care system which demands confidence and cannot exist without secrecy. The physician may only reveal information if he has the patient's consent which can also be given implied. He can be justified by a presumed consent or the balancing consideration of legally protected values. However, a duty to disclose does not exist. The medical confidentiality applies towards relatives, colleagues who are not consulted by the patient and the media. PMID- 10683891 TI - [Special problems of confidentiality in hospitals from the legal point of view]. AB - Medical confidentiality is applicable to hospitals as well. Hospital physicians are obliged to keep medical confidentiality to those responsible for the hospital, to relatives, other physicians, authorities, health insurance companies and medical services. However, this does not apply if the patient gives his consent or is presumed to give his consent or if there is a regulation of the law which justifies the violation of medical confidentiality. PMID- 10683892 TI - [Special problems of confidentiality in the hospital from the physician's point of view]. AB - Professional discretion is a result of the individual rights of the patient and the increasing right of self-determination. It is very difficult to follow these rules because of the necessity for precise documentation, electronic dommentation, and the medical and administrational necessities in the hospital. How to assure the discretion in an ambulatorium, in the room with several beds? Where to place the operation-schedules with all their details? How to assure the discretion, when a lot of people in the hospital are involved in the therapy and the diagnostic procedures? A lot of questions: everybody has to do his best, keeping in mind the professional discretion as an important human right. PMID- 10683893 TI - [Medical confidentiality towards employers,health insurance medical services and health insurance companies from the point of view of a medical society]. AB - In the Federal Republic of Germany, the medical discretion is protected by criminal law section 203, by section 9 of the medical professional law, and by the state law regarding data protection. Patient's secret may only be revealed if the patient agrees, if a law demands or allows the revelation, or if a legal good of higher value facilitates the revelation. The employer may be informed about the fact of sickness of an employed and about the results of an occupational medical screening but not about diagnosis and therapy. Under specific circumstances, the health insurance companies have the right to demand revelation from the physician if this is legally permitted or if the patient agrees. Certified hospitals have to reveal certain data to the insurance companies. Medical services of health insurances have the right to demand certain data from the physician if a health insurance company has ordered an expert witness opinion or examination and if the revelation is necessary for this examination. If it is suspected that there is a mismanagement about the numbers of beds occupied by patients in a hospital, the medical service of a health insurance may examine all patient's records related to the health insurance of interest. There exists no legal rights or duties to reveal information to private health insurance companies. The revelation of patient's data to such companies is most commonly not allowed. PMID- 10683894 TI - [Medical confidentiality towards health insurance, employment, state offices and health insurance companies--legal guidelines]. AB - The relationship between physician and patient has become multi-dimensional due to the inclusion of state offices, health insurance companies and other insurance companies. Thus, the medical confidentiality has been compromised, inevitably information will be spread into a wider circle. Nevertheless, the principle of confidentiality should be upheld. PMID- 10683895 TI - [Confidentiality and the refusal to give evidence]. AB - The physician's obligation to give evidence as a witness in a preliminary investigation is in an area of conflict between the duty to tell the truth and preservation of professional discretion. These have to be weighed in the individual case. In opposition to the Anglo-American law, where a possible right of the witness to refuse to give evidence is limited by the principle of "finding the truth", the German law of criminal procedure contains far-reaching rights of a physician and so-called professional assistants witness to refuse to give evidence in sections 53, 53a Code of Criminal Procedure in order to protect the professional secrecy. This privilege refers to all facts, that have become known to the doctor or his staff and therefore it goes beyond the area of the medical discretion in section 203 GCC, that only contains secrets, which were confined to the doctor or which have become known to him. The witness can decide whether he either uses his right to refuse to give evidence or gives evidence without being released from medical confidentiality. In the second case, he risks being punished under section 203 GCC. If a physician is considered as witness in a procedure, the medical files are protected from attachment in section 97 subsection 1 numbers 2 and 3 Code of Criminal Procedure. In cases, where the physician is defendant himself, he cannot refer to this protection. PMID- 10683896 TI - [Medical confidentiality and data protection in medical research-- legal aspects]. AB - Medical confidentiality and data protection are important for medical research. They should not impede medical progress. Unfortunately, the German legal situation is not easy to understand. There are different statutes which have to be reconciled. It is necessary that by special statute the particular problems of medical research and confidentiality as well as data protection should be regulated. PMID- 10683897 TI - [Data protection in medical networks for the point of view of a representative for data protection]. AB - The processing of medical data makes great demands on the guarantee of confidentiality, integrity and authentity. This applies to all layers of telecommunication: networks (telecommunication law), services (law of new media) and contents (data protection law). The use of strong encryption, data economicalness, offer of anonymous and pseudonym services and suitable methods of authentication are prerequisites of data medical networks which comply with data protection demands on the side of the physicians. The patients' sensitivity regarding their privacy has to be strengthened too. PMID- 10683898 TI - [Data protection in medical networks from the physician's point of view (short report)]. PMID- 10683899 TI - [Data protection in telemedicine--legal aspects]. AB - Recent developments in telemedicine have resulted in an increased flow of personalized medicinal data and therefore caused new in calculable risks and dangers to the right of privacy. This article discusses the particular problems of this issue: The patient's consent must be based on sufficient information about the intended data processing and data flow. Transmitting data into countries outside the European Community should prevail sufficient data protection according to the EC-Guideline. Particular attention mus be paid to the regulations of the new German Multimedia Legislature. PMID- 10683900 TI - [Medical aspects of telemedicine--chip cards--electronic reception and more (short report)]. PMID- 10683901 TI - [Medical confidentiality in the former DDR]. AB - In accordance with the communist ideology of the GDR, interventions in the private domain of citizens were made without any compunction. Notwithstanding, Section 136 of the Penal Code of the GDR made it a punishable offence to divulge in professional secrets. However, this regulation only simulated the right and obligation to respect professional secrets. Organs of the state continually disregarded medical confidentiality. Besides this, there were numerous statutory medical notifications which were also a severe burden to the relationship of trust between the physician and patient. The State Security Service of the GDR also constantly infringed medical confidentiality. In "research work" carried out at the "Juridical University" of the State Security Service, a thorough strategy of procuring information actually protected by medical confidentiality was ceveloped. The physician has sole responsibility in maintaining medical confidentiality. This also applies to the period of the GDR. There are examples of physicians resisting the State Security Service. PMID- 10683902 TI - [Resources survey of medicinal species from genus Aralia]. AB - This paper deals with the distribution area, growing environment and medicinal parts of 19 species and 1 variety, including 2 new species of genus Aralia. An index for identification of these medicinal species is presented. PMID- 10683903 TI - [Dormancy characteristic and inner inhibitory substances of fructus Schisandrae]. AB - It has been found for the first time that there is quite a number inhibitory substances in the testa and kernel of dormant Fructus Schisandrae. Differences have alas been found between the inhibitor extracted from the testa and that from the kernel. The fromer is stronger in inhibiting the root elongatain of Brassica sp, whereas the latter is stonger in inhibiting the seeds germination. The result shows that the dormancy of Fructus Schisandrae the inhibitors contained in the testa and kernel. PMID- 10683904 TI - [Effect of nitrogen, phosphorus and potassium on the growth of rhizome of Coptis chinensis Franch and its contents of berberine]. AB - It has been found out that the normal growth of Coptis chinensis is heavily affected and both NRA in leaves and berberine contents in rhizome are very low when seedlings are short of nitrogen, phosphorus and potassium. The plants grow especially worse and most of them soon die off when nitrogen is short. The new leaves are smaller both in number and size when phosphorus and potassium are short. Few roots can grow up and easily get old in the solution short of potassium. PMID- 10683905 TI - [Processing technology of rhizoma Arisaematis (Pinellia pedatisecta Schott)]. AB - Using the qualitative standard of Rhizoma Arisaematis stipulated in Pharmacopoeia and combining with the toxic reaction in experimental mice, the optimum amount of KAl(SO4)2.12H2O for eliminating the toxic reaction of Rhizoma Arisaematis was selected. Based on the comparison of experiments, two new technologies for processing Rhizoma Arisaematis have been established. Pilot production shows that these technologies are feasible for mass production. PMID- 10683906 TI - [Preparation method of foamed aerosol for use in vagina]. AB - The extraction of medicinal materials, compatibility of liquor and prescription of emulsion for use in vagina were observed. The result shows that through compound extraction, removal of tannic materials, decolorization and water precipitation, with propylene glycol and ethylene glycol as latent solvent and using lactic acid to adjust to pH4.5, a clarified solution can be obtained, and then with the help of Tween-spans emulisifying agent and propellant the stable emulsion is formed. PMID- 10683907 TI - [Quality criteria for bazhen shennongyin liquor]. AB - Radix Angelicae Sinensis, Radix Paeoniae Alba and Radix et Rhizoma Rhei in Bazhen Shengnongyin Liquor were identified by TLC. The contents of ginsenoside Rb1 in the preparation was determined by TLCS. Simple, accurate and reproducible, this method could be used as the quality criteria for this preparation. PMID- 10683908 TI - [Determination of emodin in kangnaoshuai capsules and in radix Polygoni multifori by thin layer chromatography scanning]. AB - The content of emodin in Kangnaoshuai Capsules was determined by TLC scanning. The method is simple and the result is accurate. The average recovery is 99.17% and RSD is 1.14%. The content of emodin in Radix Polygoni Multiflori was also determined by the same method. The method is useful in quality control of products. PMID- 10683909 TI - [Minor saponins from leaves of Panax ginseng C.A. Meyer]. AB - Five compounds were isolated from the leaves of Panax ginseng and characterized as 20(R)-protopanaxatriol, daucosterine, ginsenoside-F2, ginsenoside-F3 and majoroside-F4 on the basis of spectral analysis and chemical evidence. Among them, majoroside-F4 is obtained from plant for the first time. PMID- 10683910 TI - [Flavonoids of Cuscuta australis R. Br]. AB - Six flavonoids were isolated from the seed of Cuscuta australis and four of them were identified as kaempferol, quercetin, astragalin and hyperoside. Hyperoside was obtained from this plant for the first time. In comparison with the flavonoids in C. chinensis, it is found that quercetin and its glycoside are the main flavonoids in C. australis. This result suggests that the flavonoids can be used to distinguish these two medicinal materials. PMID- 10683911 TI - [Chemical constituents of Strychnos nitida G. Don]. AB - Six compounds were isolated from the root and stem of Strychnos nitida for the first time. On the basis of chemical properties and spectral data, the compounds were identified as beta-sitosterol, strychnine, brucine, cantieyine, lignoceric acid and palmitic acid. PMID- 10683912 TI - [Quantitative determination of neferine in plumula Nelumbinis by thin layer chromatography scanning]. AB - The content of neferine in Plumula Nelumbinis wsa determined by dual-wavelength TLC-scanning. The linear range was found to be 0.5-6.7 micrograms. The average recovery was 100.35%. The contents of neferine in growing in Fujian, Jiangxi and Hubei provinces Plumula Nelumbinis were found to be 0.199%, 2.46% and 2.48% respectively. PMID- 10683913 TI - [Screening of antiviral agents from medicinal herbs by means of Hepadnaviruses models]. AB - The antiviral study of 21 Chinese medicinal herbs was carried out in vitro and in vivo. The extracts of phyllanthus urinarin and polygonum cuspidatum exhibite obvious effects on duck hepatitis B virus and human hepatitis B virus, while the extract of Eclipta alba showed limited inhibition on HBV DAN polymerase. PMID- 10683914 TI - [Antioxidative effect of constituents of herba Epimedii (ESPS)]. AB - Herba Epimedii is a traditional Yang invigorating Chinese herb widely used in clinic. The experimental results have shown that ESPS obviously increases superoxide dismutase(SOD) activity of red cells and liver in aged mice and rats, and increases glutathione peroxidase(GSH-Px) activity of red cells in aged mice. On the other hand, ESPS obviously helps to reduce the content of serum and liver lipoperoxide(LPO) in aged mice and rats, as well as the content of lipofuscin(LF) in cardiac muscle of aged mice. The results suggest that being helpful in increasing SOD and GSH-Px activities and inhibiting the formation of LPO and LF, ESPS may be a good antiageing agent. PMID- 10683915 TI - [Pharmacological effects of zisu and baisu]. AB - The extract and volatile oil of zisu (Perilla frutescens) has shown significant antipyretic effect in rabbits and antiemetic effect in pigeons. The fatty oil extracted from its seeds has significant antitussive effect in mice and anti asthmatic effect in guinea pigs. The extract, volatile oil and fatty oil of Baisu, which is of the same genus as Zisu have the same effects as those of Zisu. The acute toxicities of the extract and fatty oil of Zisu and Baisu, whether by peroral or by intraperitoneal, are similar to each other. These results indicate that Baisu has the same pharmacological effects as Zisu, and thus can be used as a substitute for Zisu. PMID- 10683916 TI - [Characterication of reference strains of L. interrogans in China by ISSP-PCR]. AB - The gene polymorphism of the DNAs extracted from reference strains of L. interrogans in China was characterized by LSSP-PCR primered by G1 or G2, which were a pair of specific primers for L. interrogans. The fingerprinting produced by LSSP-PCR showed that serovar lai, serovar canicola, serovar pyrogens, serovar autumnalis, serovar australis, serovar linhai, serovar wolffi and serovar haemolytic have similar patterns, but serovar hebdomadis, serovar javanica, serovar ballum, serovar pomona, serovar spaidjin, serovar tarassov and serovar manahao I have different profiles. This result agreed with the classification of genetic species by Yasuda. The utilization of LSSP-PCR banding patterns in the identification of leptospries in blood samples also gained encouraging results. Compared with the routine methods in genetic species classification, LSSP-PCR has the advantages of rapidity, simplicity and low-cost. It appears to be a promising tool in studying such slowly growing bacteria as leptospires. Further exploration of LSSP-PCR in classification and identification of Leptospires is worthwhile. PMID- 10683917 TI - [Immunogenecity of expressed protein p68 from recombinant plasmid rpDJt in L. interrogans serovar lai]. AB - There are two types of infection caused by pathogenic microorganisms, intracellular infection and intercellular infection. Infection of pathogenic leptospira is an intercellular infection. The immunological reaction of host to intercellular infection is unique. The potential immunogen of an expressed protein should meet three criteria: it can be degraded (by antigen-present cells in the host); it should have antigenic epitope which can be recognized by specific antibodies and have at least one epitope that can be recognized by an MHC II protein and T cell receptor. In this study we report the cloning of an L. interrogans protein in plasmid rpDJt and the immunogencity of the expressed protein derivative. A genomic library of L. interrogans serovar lai strain 017 was constructed with the plasmid vector pUC18. Recombinant plasmids, designated pDJH2 and pDJ8 were screened from the bank. EcoRI-inserted fragment of 1. 9 kb recombinant DNA of pDJH2 was ligated into T7 RNA polymerase/promoter vectors (pT7 7). Then they were transformed into E. coli JM109 (De3), one of subclones, designated rpDJt was achieved. SDS-PAGE showed that the molecular weights of expression proteins were 68 kd and 23 kd respectively, designated p68 and p23. Purifying and isolating p68 and p23, we separated them from SDS-Polyacrylamide gels by using Side-Strip method. After fragmenting and electroeluting, p68 and p23 were injected into guinea pigs and rabbits. An extremely strong immune response to p68 was obtained since an anti-p68 antibody response could be detected to a dilution 1:524,288 (guinea pigs) and 1:262,144 (rabbits) by ELISA while anti-P23 antibody being 1:1024 (the same to guinea pigs and rabbits). The results of improved MTT and conA 3HTdR transformation methods showed the activities and proliferation of Th-cells were increased in guinea pigs after p68 immunization (IL-6, 83.25 IU/ml, IL-2, 28.75 IU/ml; RPI, 2.04, SI, 65.62%) Thlymphocyte existed in two subclasses, the Th1- and Th2-cells. A major role of Th2-cells is to "help" B-cells differentiate, replicate, and secrete antibody. The properties of these interactions explain why p68 makes good antigen and p23 does not. The antigens responsible for eliciting the production of protective antibodies are not known; however, several outer membrane proteins on L. interrogans are candidates for vaccine. Our results suggest that expresion protein p68 from recombinants (rpDJt) may be a candidate for gene engineered subunit vaccine for Leptospirosis. PMID- 10683918 TI - [Characterization of immunoblots of hydrophobic outer membrane proteins Leptospira interrogans serovar lai strain 017]. AB - Outer membrane proteins (OMP) of Leptospira interrogans serovar lai strain 017 were extracted by using Triton X-114 (TX-114). The OMP were solubilized and phase partitioned into both the hydrophilic, aqueous phase and hydrophobic, detergent phase. TX-114 did not solubilize the protoplasmic cylinder in intact organisms. The protoplasmic cylinders contained a lot of protein bands and most of the TX 114 solubilized proteins partitioned into the aqueous phase, whereas only 14 protein bands entered the detergent phase with SDS-PAGE. Detergent phase proteins were of 5 major protein bands such as 66 kd, 39 kd, 35 kd, 27 kd, and 16 kd. Immunoblotting of the material extracted with TX-114 showed that detergent phase proteins of alone 39 kd was apparently immunoblotting with antiserum against such as whole cell of 017 strain the outer envelope of 017 strain and the immunoprotective anti-017 Mb E4B7G5. The results showed that we could separate and purify the 39 kd protein to analyse the amino acid sequence for the cloning, expression and development of genetic engineering vaccines. PMID- 10683919 TI - [Analysis on speciems of serum and urine in 66 cases of early leptospirosis by PCR and biotin-AMPPD hybridization]. AB - We used polymerase chain reaction (PCR) and Biotin-AMPPD hybridization to detect leptospiral DNA in blood and urine samples in 66 patients at the early stage of leptospirosis. The results showed that PCR and Biotin-AMPPD hybridization not only ruled out the non-specific PCR amplification and increased the reliability for clinical specineus of leptospirosis but also raised the detecting sensibility (from 71.3% to 86.1%). The positive rates of PCR resulted from serum and urine showed no statistical difference; therefore urine sample is worthy of application and dissemination for detecting leptospires at the early stage of leptospirosis. Urine sample is easier for one to collect, preserve and has less intervention. The primers G1, G2 are optimal opplication of PCR in epdemic areas in China. PMID- 10683920 TI - [Alterations of substance P-immune reaction positive neurons of cerebral tissues in epileptic rats]. AB - Employing the immunocytochemical analysis, we observed the alterations of Substance P-Immune Reaction (SP-IR) positive neurons of the cerebral cortex, hippocampus and amygdala in rats suffering from epilepsy induced by Penicillin (PEN). The result showed that the number of neurons of epileptic group was higher than that of the control group and no significant change in the rumber of neurons was observed in the group in which PEN was given after injection of Nimodine. It indicates that SP and Ca2+ participate in the process of epileptogenesis. PMID- 10683921 TI - [Regeneration of functionally active rat brain muscarinic receptor in vitro after inhibition with methylmercury chloride]. AB - The effect of methylmercury on muscarinic receptors and the regeneration of functionally active muscarinic receptor in vitro by antagonists were investigated. The result showed that methylmercury chloride (MMC) inhibited the binding of [3H] QNB to muscarinic receptor of rat brain-lysed synaptosomes, with IC50 values of 4.18 mmol/L. Regeneration of functionally active rat brain muscarinic receptors after inhibition with methylmercury was achieved by 2,3 dimercapto-1-propanesulfonc acid, Na salt (DMPS) dithiothreitol (DTT), glutathione (GSH) and Cysteine. Blocking the sulfhydryl groups is suggested to be the molecular mechanism of inhibition of brain muscarinic receptors by methylmercury. Our results provide evidence that thiols chelate out mercuric cations that tightly bound to sulfhydryl groups in muscarinic receptor binding sites and regenerate [3H] QNB binding activity. PMID- 10683922 TI - [The effects of Tripterygium wilfordii polyglycosidium on neuromuscular junctions of adult toad]. AB - The effects of tripterygium wilfordii polyglycosidium (TWP) on the transmission at the neuromuscular junctions (NMJ) of sciatic nerve-sartorius muscle preparations of 20 adult toads (Bufo bufo gargarizans) in vitro were studied by means of intracellular microelectrode recording of endplate potentials (EPP). The results revealed that TWP could cause changes of EPP amplitude so as to influence the transmission at NMJ, and the effects apparently depended on the concentration of the drug. A proper concentration of TWP could produce a significant increase in EPP amplitude and therefore facilitate the transmission at the NMJ. PMID- 10683923 TI - [Application of Apo B 3' DNA polymorphisms in forensic science Practice]. AB - Amp-FLP analyses of Apo B 3' VNTR locus in human blood stain, saliva (stain), semen, mixed stain and hair have been carried out. The genotypings have been achieved. The genotypes of Apo B 3' locus can be detected accurately in blood stains kept at room temperature within 15 weeks and at -20 degrees C within two years. In a series of paternity testing on 34 cases, 8 false alleged fathers were excluded; of them, 6 were excluded by Apo B 3' locus alone or by Apo B 3' locus combined with other genetic markers. 3 rape cases were investigated; 2 suspects were excluded. PMID- 10683924 TI - [The application of D1S80 locus analysis to forensic problems by using Amp-FLP technique]. AB - This is a paper on paternity testing in 85 test cases and the identification of individuals from mixed stains of semen and vaginal secretion in two rape cases. We used primers flanking the hypervariable region of the D1S80 gene to amplify DNA extracted from fresh blood, blood stain, saliva stain, semen stain, mixed stain in vaginal swab, and from fetal tissues. The results revealed that 35 out of 85 alleged fathers in 85 paternity testing cases were excluded. In 30 of the 35 excluded cases, the alleged fathers were excluded by D1S80 locus analysis. The exclusion rate was 35.29%. In 2 of the excluded cases, the alleged fathers were excluded by D1S80 locus analysis only. In 50 nonexcluded paternity testing cases, the paternity index(PI) of D1S80 locus was 2. 0576-111.1111. It indicates that D1S80 locus analysis plays an important role in elevation the relative chance of paternity (RCP) value. In 41 of the 50 nonexcluded cases, RCP was higher that 99.75%. RCP reached 99.300%-99.649% in 9 of cases. In one rape and homicide case, the genotype of D1S80 locus of sperm on the vaginal swab was detected. It gave a clue to investigation. In the other rape case, the genotype of D1S80 of sperm on the vaginal swab was consistent with the blood stain from the suspect. So the suspect was not excluded. Since the D1S80 locus has a high discrimination power it is very useful for forensic individual identification and parentage testing. Some problems have been discussed in this paper. PMID- 10683925 TI - [Transferrin subtyping of human semen, semen stain, vaginal fluid and mixed stain using isoelectric focusing and immunoblotting]. AB - Transferrin (Tf) is an important genetic marker for personal identification in forensic science. We examined the subtypes in human semen, semen stain, vaginal fluid and mixed stain using isoelectric focusing and immunoblotting method(IEFIB). The results showed that, Tf subtypes of 8 semen and semen stain samples stored at room temperature for up to 32 weeks were the same as those in sera. Tf subtypes of 17 vaginal fluid and 4 mixed stain containing few semen were not detected. Tf Subtyping of 1 mixed stain containing enough semen was successful by using IEFIB. These results indicate that Tf is stable in semen stain. The Tf subtyping in semen stain and mixed stain containing enough semen can be carried out by using IEFIB method. PMID- 10683927 TI - [Optimazation of conditions for stereoselective reaction by enzyme-catalyzed acetylation]. AB - Optically active endo-tricyclo-[5,2,1, 0(2,6)]deca-8-en-3,5-diol-5-acetate is a very useful chiral synthon for the synthesis of optically active natural products. In this paper, the optimal conditions for the synthesis of this compound was established by orthogonality test. Under these conditions, the chemical percentage yield of acetylation of (+) endo-tricyclo-[5,2,1,0(2,6)] deca 8-en-3,5-diol-5-acetate and its % e.e. can reach up to 81% and 98.3% respectively. The optimazation of the conditions is efficient for the reaction of enzyme-catalyzed esterification. PMID- 10683926 TI - [Study of PEFV curves in 520 normal middle-aged and old humans]. AB - Partial expiratory flow-volume(PEEV) curves and other pulmonary functions were performed for 520 (20-86 yr) normal midde-aged and old humans. The regression equations of V at FRC (VFRC, volume corrected VFRC (VFRC/FVC), flows at various lung volumes (V75, V50, V25) and specific time constants (S tau 75, S tau 50, S tau 25) were built with multiple stepwise regression analysis. VFRC decreased with increase of age, but after correction with FVC, the correlation of VFRC/FVC with age decreased. The regressions were tested with substitution of parameters with normal persons and proved to be correct. This study suggests that PEFV curve with convenient method has theoretical and practical significance. PMID- 10683928 TI - [Callus formation of Aristolochia tuberosa and determination of aristolochic acid of callus by HPLC]. AB - Studies on the callus induction starting from the young stems of Aristolochia tuberosa on MS medium were reported and the morphological characteristics of cultured cells were described. The results of HPLC determination showed that some callus cultures had higher aristolochic acid content than their original plant organs, and the callus cultured in MS medium +2, 4-D 0.5 mg/L produced more amounts of airstolochic acid(0.27% of dry callus weight) compared to the original plant organ (0.15/1000 of dry weight). PMID- 10683929 TI - [Mutagenicity and carcinogenicity of nabumetone]. AB - To study the mutagenicity and carcinogenicity of Nabumetone, we conducted Ames test (TA97, TA98, TA100, TA102), micronucleus test(MN) in mice marrow, chromosomal aberration assay(CA) in CHL cells in vitro, CA in germ cells from testes of mice, and cell transformation test of Syrian hamster embryo(SHE) cells. The maximum concentration was 500 micrograms/plate in Ames test with and without S9 mix. The mice were treated orally(gavage) daily for 4 days in 3 doses in which the maximum dose was 60% LD50 and sampled at the 5th day in MN. The maximum concentration was the dose that the growth of 50% of cells was inhibited in CA of CHL. Cells were harvested after recultured for 18 hours in fresh medium after treatment for 6 hours in the test with S9 mix, and after treatment for 24 or 48 hours in the without S9 mix. The mice were treated orally(gavage) daily for 5 days in 3 doses in which the maximum dose was 1/4 LD50 and sampled at the 6th day in CA of germ cells from testes of mice. 2 micrograms/ml was chosen as the maximum concentration in the cell transformation test of SHE cells, and result was observed after treatment for 9 days, All the tests obtained the same negative result as that reported by other investigators. PMID- 10683930 TI - [Antiallergic effects of tranilast in rats and guinea pigs]. AB - Tranilast is an anti-allergic drug. In this study, we made a comparision between the Tranilast synthized by School of Pharmacy WCUMS using new technical and the Tranilast produced by Kissei pharmaceutical Co. LTD, Japan on their antiallergic effects. We found that the two tranilasts had the same antiallergic effects: (1) they inhibit the passive cutaneous anaphylaxis in sensitized rats with the dose of 100, 200 mg/kg(P < 0.01); (2) they inhibit degranulation of mast cells in sensitized rats (10(-5) and 10(-4) mol/L) (P < 0.05); (3) they inhibit schultz Dale response in sensitized guinea pigs (10(-3) and 10(-4) (mol/L); (4) the inhibit SRS-A release from the lung of sensitized guinea pigs(10(-3), 10(-4) and 10(-5) mol/L) (P < 0.05); and (5) they inhibit the contraction of ileum of normal guinea pigs induced by SRS-A(10(-4) and 10(-3) mol/L). PMID- 10683931 TI - [Molecular cloning of the imipenem resistant gene in bacteroides fragilis]. AB - Imipenem, as a representative of-cabapenem, is one of the most effective beta Lactam agents against Bacteroides, but recently there have been a few reports on the resistant strains of Bacteroides. To explore the mechanism of imipenem resistance (IMPr) at the molecular level, we extracted chromosome DNA of the resistant strains and acquired a 1.7 kb fragment of resistant gene of imipenem. Recombinant plasmid was stably reserved in generation after generation in E. coli DH5. Resistant gene of imipenem was labeled with Dig-11-dUTP by way of random prime, and 40 strains of clinically isolated Bacteroides were detected by the prepared DNA probe. Two IMPr strains showed hybridization signal, whereas 38 IMPr strains did not show such a signal. The result suggests that the Dig-labeled IMPr gene probe can be used in molecular epidemiology for investigating IMPr strains. PMID- 10683932 TI - [The responses of leukemia cells to interleukin-2 in vitro]. AB - We conducted a study on the utility and safety of rhIL-2 immunotherapy for leukemia. The responses of leukemic cells expressing interleukin-2 receptor (IL 2R) gene obtained from 7 leukemic cell lines, 6 cases of acute lymphoblastic leukemia (ALL) and 8 cases of acute myeloid leukemia (AML) were measured by 3H thymidine incorproation. The results showed that the responses of these cells were heterogeneous. The cells expressing IL-2R alpha and IL-R beta mRNAs simultanesously from NKL-1,2 cases of ALL and 1 cases of AML(M5) responded to rhIL-2 proliferatively; the cells from 3 leukemic cell lines, 2 cases of ALL and 2 cases of AML were inhibited by rhIL-2; the leukemic cells from the remaining 3 leukemic cell lines and 7 cases of leukemia did not respond to rhIL-2. The study demonstrates that the responses of leukemic cells expressing IL-2 receptors to rhIL-2 are heterogeneous. The patterns of the responses are proliferative, being inhibited to rhIL-2, or irresponsive. So rhIL-2 immunotherapy is suitable for the patients whose leukemic cells were inhibited by or did not respond to rhIL-2 in vitro, but this immunotherapy is not suitable for the patients whose leukemic cells responded proliferatively to rhIL-2 in vitro. PMID- 10683933 TI - [Properties of adherence of erythrocytes to endothelial cells in patients with ischemic stroke]. AB - Adherence of erythrocytes to human umbilical vein endothelial cells (HUVEC) in patients with cerebral thrombosis and transient ischemic attack (TIA) were quantitatively studied in the flow chamber system, compared with that in healthy subjects. We found: (a) The adherence of erythrocytes to HUVEC in patients with cerebral thrombosis and TIA increased, compared with that in healthy subjects; (b) The adhesion of erythrocytes to HUVEC in patients with TIA was not significantly different from that of patients with cerebral thrombosis. These suggested that the increased adhesion of erythrocytes to endothelial cells might play an important role in the pathologic process of ischemic stroke. PMID- 10683934 TI - [Evaluation of oral glucose tolerance test in the assessment of reserved function of liver for patients with hepatocellular carcinoma]. AB - The aim of this study was to evaluate oral glucose tolerance test(OGTT)in the assessment of reserved function of liver for predicting the tolerability of patients to hepatectomy and hence provided a criteria for selecting the candidates for undergoing hepatectomy, since the majority of hepatocellular carcinoma (HCC) patients were associated with posthepatitis cirrhosis. The preoperative and postoperative OGTT and liver biopsy for pathological investigation were carried out in 62 cases of hepatecomized patients and 49 cases of unresected patients for comparison. The results revealed that the patients whose preoperative OGTT curve was of P type recovered uneventfully after hepatectomy, but those whose curve was of L type of tolerated poorly to hepatectomy and were liable to postoperative hepatic failure and complications. The severity of cirrbosis in those poor risk patients fell to C III or C IV histological degree. 29 patients with intermediate feature of OGTT curve between P type and L type, i.e. I type underwent regional vascular occlusion at hepatic hilus as hepatectomy, and infusion of Danshen extract solution before vascular occlusion to prevent hepatocytes from reperfusion injury. Of them, 20 recovered uneventfully, 8 suffered from complications such as ascites and/or juandice, and 1 died within 1 month after operation. The followup study showed that the survival time of patients with P type OGTT curve was longer than that of I type, and the latter was longer than that of L type. The pattern of OGTT curve could change from preoperative P type to postoperative L type, depending on the severity of vascular interruption of liver and the ischemic injury to hepatocytic mass in operation. PMID- 10683935 TI - [Kidney lesions complicated by hypoxic ischemic encephalopathy of neonatal pig]. AB - To study the kidney lesions complicated by hypoxic ischemic encephalopathy (HIE) in newborn, we successfully established a neonatal pig model of HIE. Changes in brain tissue similar to those of newborn in HIE were detected after 2 hours of hypoxia and ischemia. Meanwhile, swelling of kidneys, hemorrhage below encapsules, swellin of tubular epithelial cells, necrosing and falling of a few tubular epithelial cells, and narrowing of tubular lumens were observed. The basement membranes of renal tubules remained normal serum BUN and creatinine increased significantly after experiment compared with before (P < 0.025 and P < 0.05 respectively). The results showed that the kidney of neonatal pig was damaged in HIE. But the injury was not severe. After 72 hours free from hypoxia, the structure and function recovered approximately from the injury. PMID- 10683936 TI - [Medicine S inhibits class II MHC expression of intragraft renal in rats]. AB - The Purpose of this study was to address the mechanism of medicine S which has anti-rejection effects of renal allografts in rats. Kidney transplantations were performed from SD to Wistar strain (allogeneic) and from Wistar to Wistar (Isograft) using the same modified technigue described by Fabre and kamada. Experimental rats were divided into five groups. Group I (Isograft group) and group II (allograft group) as controls were not treated with medicine. The others were allograft groups which received medicine S, Cyclosporine A, and low-dose Cyclosporine combined with medicine S, respectively. Renal function and resultant morphology changes were assessed 2, 4 weeks after transplantation. All sections of kidney grafts were stained with monoclonal antibody class II MHC (OX6), and then the surface densities of positive staining were quantified by computer image analysis. The level of molecular expression in group II was significantly increased (7.61 +/- 0.57 vs 0.51 +/- 0.2 of group I, P < 0.01). In groups I and IV, the molecule of expression was reduced, compared with the groups II, III and V (P < 0.05). The results suggest that medicine S decreases the level of class II MHC expression and medicines combined with lowe-dose cyclosporine is more effective than cyclosporine alone. PMID- 10683937 TI - [Correlations of T lymphocyte subsets and IL-2, sIL-2R in patients with malignant ovarian tumor]. AB - To study the relation between malignant ovarian tumor (MOT) and cell mediated immunity, the numbers of T lymphocyte subsets and levels of IL-2 and sIL-2R were measured simultaneously in 40 patients with MOT and 30 normal controls (NC), and the correlations between T subsets and IL-2, T subsets and sIL-2R were analyzed. The results were as follows: (1) The numbers of CD3+, CD4+, CD4+/CD8+ and the levels of IL-2 in the patients were lower than those in the NC significantly, and the opposite results were observed about CD8+ and sIL-2R (P < 0.01-0.001); (2) There were positive correlations between CD3+, CD4+, CD4+/CD8+ and IL-2 and there were negative correlations between CD3+, CD4+, CD4+/CD8+ and sIL-2R in MOT group (P < 0.01-0.001). The results suggested that the T lymphocyte immune function in MOT group was decreased. The reduced numbers of CD3+, CD4+, CD4+/CD8+ were related to the decreased levels of IL-2 and the increased levels of sIL-2R. It may be an important factor of incidence and development in MOT. PMID- 10683938 TI - [Relationship of C-erbB-2 oncogene overexpression to estrogen progesterone receptors in brease cancer and its prognostic significance]. AB - In order to understand the relationship of C-erbB-2 oncogene overexpression to ER and PR in breast cancer and its prognostic significance, we examined overexpression of C-erbB-2 oncogene in 106 breast carcinomas by using immunohistochemical techniques (LSAB). The results showed that the positive rate of C-erbB-2 overexpression was 63.21% (67/106). The overexpression of C-erbB-2 oncogene related negatively with survival. 81.63% of the cases with overexpression of C-erbB-2 oncogene survived < or = 5 years and 34.29% survived > 10 years. There were significant associations of C-erbB-2 overexpression with advance clinical stage, high histological grade, and positive axillary node status in breast cancers. Negative relationship between hormone receptors and C erbB-2 oncogene. All of these findings suggested that overexpression of C-erbB-2 oncogene might be an important prognostic factor and the detection of C-erbB-2 oncogene might be arranged as a regular pathological examination in the cases of breast cancer. PMID- 10683939 TI - [Effect of haemorrheological changes on acute pancreatitis]. AB - This study sought to determine the effect of the haemorrheological changes on the pathological damage to pancreas in acute pancreatitis. 96 Wistar rats, four months old, were allocated into three groups: group I (n = 32) received surgery for pancreatic duct obstruction (PDO) with secretion stimulation; group II (n = 32) for PDO with high molecular dextran (DX110) injection intravenously, and group III (n = 32) for PDO with secretion stimulation and DX110 injection intravenously. Ten other rats were used as controls (group IV) for laparotomy alone. The results showed that PDO with hypersecretion could induced edematous pancreastitis and PDO with DX110 injection induced only very lightly oedema in the pancreas which was similar to the result of PDO alone, although the haemorrheological changes were obvious in these rats. PDO with hypersecretion and DX110 injection induced acute necrotic pancreatitis, and the pathological lesion in the pancrease which changed gradually from edematous to necrotic could be observed. This result suggests that haemorrheological change may not be a causative factor of the acute necrotic pancreatitis, but it probably could exacerbate the damage to pancreas in acute pancreatitis and play an important role in the transformation from edematous to necrotic pancreatitis. PMID- 10683940 TI - [Diagnostic value of pleuro-examination with fiberoptic bronchoscope in indefinite pleural effusions]. AB - The diagnostic value of pleuro-examination with fiberoptic bronchoscope in indefinite pleural effusions was studied. Pleuro-examination with fiberoptic bronchoscope has been performed in 10 cases with indefinite pleural effusions, and its diagnostic value and complications have been observed. The results showed that the diagnoses were confirmed in 9 of 10 patients submitted to pleuro examination, and no severe complications were found. The results suggest that pleuro-examination with fiberoptic bronchoscope is an effective and safe procedure and might be performed on patients with pleural effusions when routine examinations fail to provide a diagnosis. PMID- 10683941 TI - [Determination of trace silver in surface water samples by flame atomic absorption with continuous flow on-line absorption preconcentration]. AB - In a continuous flow on-line preconcentration system, trace silver was adsorbed onto activated carbon particles packed in micro colum at pH 1, eluted with 0.3 ml of 10 g/L Na2S2O3 solution and determined by flame atomic absorption spectrometry. For surface water samples, 25 ml sufficed precise determination of silver at level of microgram/L. The relative standard deviation of 10 parallel sample determinations was 6.9%. The proposed method was successfully applied to water samples with recoveries ranging from 93.2% to 104.6% and a throughout of 7 8 sampling/h. PMID- 10683942 TI - [A simple and rapid method of preparing cell or tissue embedded in situ for transmission electron microscopy]. AB - A simple and rapid method of preparing cell or tissue embedded in situ for transmission electron microscopy (TEM) is recommended. It is very simple; all procedures, including fixation, dehydration, infiltration and embedding, are performed on the same carrier (culture vase or microscope slides). It is also very rapid; the whole process takes only five hours. The entire cell or tissue embedded in situ is easily separated from the carrier. The ultrastructures of the cell or tissue embedded in situ are well preserved with minimal damage. This method can be used in preparing cultured monolayer cell for TEM study, paraffin section for electron microscopic diagnosis, and frozen section for TEM immunohistochemistry or enzymehistochemistry. The main points of the procedures of preparation have been discussed. PMID- 10683943 TI - [The compressive strength, tensile strength, flexural strength and micro-hardness of Plat-II Castable Ceramics]. AB - In preparation for the clinical use of Plat-II Castable Ceramics (PCC-II), we tested its compressive strength, tensile strength, flexural strength and microhardness. The flexural strength was tested by the three-point bending test. The result showed that the compressive strength was 541.7 MPa; the tensile strength 42.5 MPa; the flexural strength 142.0 MPa; the modulus of elasticity 61.4 GPa; and the micro-hardness 499.6 kg/mm3. The strength anol micro-hardness of PCC-II materials were better than those of human enamel. So PCC-II is applicable to restorations in dentistry. PMID- 10683944 TI - [The immune response in rats immunized systemically by the surface protein antigen P1 from streptococcus mutans conjugated with procholeragenoid]. AB - This study was carried out to observe the antibody responses in rats after they were immunized with the surface protein P1 of streptococcus mutans when a special adjuvant was used. Antigen P1 was conjugated covalently with procholeragenoid (PCG), and then Sprague Dawley rats were immunized with P1 or the conjugated antigen P1-PCG subcutaneously or intragastrically. Anti-P1 antibody level was assayed at different time points by ELISA. The results showed that the levels of anti-P1 SIgA antibody in saliva rose when P1-PCG was given subcutaneously or intragastrically; the antibody level following the subcutaneous injection was higher and lasted longer, compared with that following the intragastric administretion. The level of anti-P1 IgG antibody in serum only rose when the rats were immunized subcutaneously. These results implied that mucosal immune response or humoral immune response could be induced subcutaneously when PCG was used as an adjuvant. PMID- 10683945 TI - [The influence of porcelain thickness and non-uniformity on porcelain cracks in implant-supported metal-porcelain fixed bridge]. AB - This experiment studied the influence of porcelain thickness and non-uniformity on porcelain crack in implant-supported metal-porcelain fixed bridge. The result indicated that porcelain crack began to appear when the body porcelain powder was 3-5 mm in thickness; more cracks took place when the powder thickness increased by 1 mm to 2 mm on the axial surface in the axial direction; cracks became serious when the thickness suddenly changed to zero; and connectors were liable to cracks. Therefore, in designing and fabrication, one should avoid and sudden change in the shapes of the connector and the porcelain on it, any sudden change in the thickness of porcelain and a thickness of porcelain powder not less than 3.5 mm. Other-wise, it is neccessary to use internal crown between metal base and abutment to meet the demands. PMID- 10683946 TI - [Change of serum lipid, apolipoprotein during cholesterol gallstone formation in rabbit model]. AB - In order to study the formation of cholesterol gallstone through rabbit model which was induced by high cholesterol diet (HCD), we investigated the rabbits' serum lipoprotein cholesterols and apolipoprotein (apo) at 1 week (1w), 2 weeks (2w), 3 weeks (3w) and 4 weeks (4w) in comparison with those of a control group respectively. The results were as follows: (1) of 10 rabbits subjected to experiment, 4, 6, and 7 rabbits were found to have induced-cholesterol gallstones in the 2w, 3w and 4w groups respectively. (2) The serum concentrations of total cholesterol (TC), triglyceride (TG), phospholipid (PL), low density lipoprotein cholesterol (LDL-C) and very low density lipoprotein chloesterol (VLDL) increased significantly (1w, 2w, 3w and 4w groups vs control group, P < 0.05), especially in the 3w and 4w groups; the surum concentrations of high density lipoprotein cholesterol and its subfractions (HDL-C, HDL2-C, HDL3-C) decreased slightly (vs control group, P > 0.05). (3) The serum contents of apoB100, apoC II and apoC III increased significantly (vs control group, P < 0.05), especially in the 3w and 4w groups; the serum apoA I reduced gradually in 1w, 2w and 3w groups, and decreased greatly in 4w group (vs control group, P < 0.05). The results demonstrated that owing to the intake of high cholestrol diet with the passage of time, the increased concentrations of serum VLDL-C, apoB, apoC II and apoC III possibly caused an enhanced secretion of biliary cholesterol into bile; that the decreased serum apoA I level might reduce the secretion of anti-nucleating factor into bile. All of these factors and changes may play important roles during the formation of cholesterol gallstones. PMID- 10683947 TI - [Penetration of ciprofloxacin and cefoperazone into human pancreas]. AB - Major pancreatic infection is responsible for more than 80% of deaths in patients with acute pancreatitis. Therefore, the role of antimicrobial drugs in the prevention and treatment of secondary parcreatic infection is very important. The choice of antimicrobial drugs must be based upon the ability of the drug to exceed the therapeutic concentration in pancreas for the common pathogens. The penetration of ciprofloxacin and cefoperazone into pancreas was investigated in ten patients who had undergone pancreatoduodenectomy. The pancreatic juice was temporarily diverted to the exterior via a panoreatic duct catheter. The pancreatic tissue was obtained intraoperatively and pancreatic juice was drained postoperatively. The antimicrobial drug concentrations were determined by high performance liquid chromatography. The concentrations of ciprofloxacin and cefoperazone in pancreatic juice were 44% and 17%, respectively, of those in serum, and exceeded the in vitro concentration (MIC-90) for most bacteria associated with pancreatic infections. The result indicates that ciprofloxacin and cefoprazone appear to be appropriate for both prophylaxis and therapy of secondary pancreatic infections. PMID- 10683948 TI - [Detection of the level of serum antigen recognized by McAbGB2 in breast cancer]. AB - This study sought to verify the levels of serum antigen recognized by McAbGB2 (abbr. serum McAbGB2 antigen) in patients with breast cancer. We adopted the ELISA method and used McAbGB2 to detect the level of serum McAbGB2 antigen in 50 normal persons and 60 patients with breast cancer. The results showed that the level of serum McAbGB2 antigen of 17 preoperative patients with breast cancer was significantly different from that of the normal persons (P < 0.001); the rate of agreement on positivity for the 17 preoperative patients with breast cancer was 88.2% (15/17); the levels of serum McAbGB2 antigen in 8 patients among the 17 preoperative patients with breast cancer decreased from 57.5 +/- 51.3 u/ml (preoperation) to 20.6 +/- 4.98 u/ml (postoperation; the levels of serum McAbGB2 antigen in the other 43 operated patients with breast cancer almost remained normal during post-operative chemotherapy. These suggest that serum McAbGB2 antigen is a marker for breast cancer and can be used for serological diagnosis, treatment and prognosis inspection. PMID- 10683949 TI - [Investigation of C1R gene frequencies in three Han populations in China]. AB - To reveal the C1R polymorphism in Chinese, three Han populations in Guangzhou (101 samples), Jilin (105 samples) and Chengdu (111 samples) were investigated with a technique using PAGIF followed by immunoblotting. The results showed in Chengdu the C1R * 1 = 0.5676, C1R * 2 = 0.3424 and C1R * 5 = 0.0856, in Guangzhou C1R * 1 = 0.5248, C1R * 2 = 0.2663 and C1R * 5 = 0.1089, and in Jilin C1R * 1 = 0.5381, C1R * 2 = 0.2619 and C1R * 5 = 0.1714. Three rare genes C1R * 6, C1R * 7 and C1R * 8 were found in the investigation. These indicate that the frequency of C1R * 2 is elevated from north to south which may imply a geographic cline in this locus. The cumulated heterogeneity of C1R in Han population is 61.5% which means that this polymorphic system is useful in anthropolgy as well as in forensic science. PMID- 10683950 TI - [p53 gene mutations in BALB/c 3T3 cells transformed by crocidolite]. AB - This study sought to address the relationship between crocidolite and p53 gene mutation. The mutations of p53 gene in 8 BALB/c 3T3 cell lines transformed by crocidolite were analysed. Altogether 11 exons of the gene from 8 cell lines were detected by PCR-SSCP. 7 alterations were found; 2 of them were located in exon 4 6, and 5 in 9-11. Most of the mutations (5/7) were of one more band than that of wild cell from SSCP, and alterations were randomly scattered among the crocidolite doses groups. The results suggest that the presence of a p53 alteration is not related to the dose of crocidolite used. Besides, p53 mutation may occur in a relatively later period of the growth of the transformed cell lines. The results also showed that the mutations occurred predominantly in exons 9-11. This was different from that seen in human mesothelioma where mutations in the exon 5-8 of p53 gene were more frequently observed. PMID- 10683951 TI - [Expression of vasoactive intestinal peptide receptor in human colonic carcinoma cell membranes]. AB - To evaluate the expression of vasoactive intestinal peptide receptor in colonic carcinoma cell membranes in men and to assess the relationship between the receptor characteristics and the histopathologic features, the authors labelled vasoactive intestinal peptide and measured vasoactive intestinal peptide receptor sites with radio-ligand bind assay on 12 specimens from colon cancer and its adjacent normal tissues. The number of the vasoactive intestinal peptide receptor sites of colon cancer was significantly smaller than that of the adjacent normal tissues (2.66 +/- 3.84 pmol/mg versus 10.54 +/- 17.99 pmol/mg, and the number of the receptor sites of colon cancer was not well related to the degrees of cancer differentiation. This study has laid the basis for the regulation of the receptor ligand binding to inhibit the growth of colon cancer. PMID- 10683952 TI - [Antiepileptic effects of nimodipine on penicillin-induced seizures in rats]. AB - Injecting penicillin (PNC) intraperitoneally in Wistar rats, we observed their epileptic behaviour and electroencephalographys. The results showed that nimodipine (NIM), a calcium antagonist, could inhibit the seizures and epileptic discharges significantly. No epileptic action was noted while NIM was injected before PNC. It supports our consideration of NIM as a new sort of nonsedative anticonvulsant in clinical practice. PMID- 10683953 TI - [Effect of tetrandrine on pulmonary vascular morphology in rats with hypoxia pulmonary hypertension]. AB - To investigate the effect of Tetrandrine on pulmonary vascular morphology in rats with hypoxia pulmonary hypertension. We established the model of hypoxia pulmonary hypertension and observed the small pulmonary arterial morphologic changes under light microscope and electron microscope after the rats were treated with Tetrandrine; also, we made morphometric analysis. The results showed that the small pulmonary arteria in rats with hypoxia pulmonary hypertension was thinner than normal ones and had proliferation of smooth muscular cells. Morphometry displayed that the external diameter became smaller and that the ratio of vascular wall thickness to external diameter (M/T%) and the ratio of vascular wall area to total area (MA%) increased. The pathologic change significantly decreased after the rats were treated with Tetrandrine, indicating that Tetrandrine inhibited hypoxia-induced thickening and muscularization of small pulmonary arteria by inhibiting the proliferation of collagenous fibers so that narrowing of small pulmonary arteria was significantly slackened and pulmonary hypertension was alleviated. Therefore, we conclude that tetrandrine may partly prevent the development of pulmonary hypertension. PMID- 10683954 TI - [The changes of glutamate receptor and free Ca2+i in hypoxic-ischemic cerebral injury: experimental study]. AB - This experiment was designed to explore the pathogenesis in hypoxic-ischemic encephalopathy (HIE). Sixteen newborn pigs were divided into two groups: (Group A) normal control and (Group B) HIE 24 hours. The glutamate receptor (Glu R) in forebrain crude synaptic membrane (SPM) and free Ca2+i in RBC were tested respectively. The results revealed that the binding sites (Bmax) of Glu R in Group B was much lower than that in Group A, but the affinity (Kd) showed no statistic difference between Group A and Group B. In addition, free Ca2+i of RBC in Group B was much higher than that in Group A. This study demonstrates that the changes of Glu R and Ca2+i are involved in the pathogenesis of hypoxic-ischemic cerebral injury in newborn animals. PMID- 10683955 TI - [Identification of the anti-rIL-1ra McAb]. AB - Using dot-immuno-blot and Western blotting techniques, we have identified the specificity of anti-rIL-1ra McAbs. The results showed that the anti-rIL-1ra McAb 2E4 specifically reacted with rIL-1ra, from which the recognizing signal of dot immunoblot was significantly positive (+2) and a single rIL-1ra protein band (20 kd) was clearly recognized by Western blotting. The control results of PcAb recognization have further proved the high sensitivity of Western blotting and high specificity of rIL-1ra McAb 2E4. The successful identification of rIL-1ra McAb 2E4 will be helpful to further understand the biologic effects, especially the pro-inflammatory properties, to study the change of rIL-1ra level during inflammation, and to instruct the clinical use of rIL-1ra. PMID- 10683956 TI - [Bone histomorphometric changes in ovariectomized goat at different time courses]. AB - Using histopatholoical method, we investigated the bone histomorphometric changes in ovariectomied goats at different time courses. Eight goats, female, aged one and a half years, were randomly divided into two groups, the control and the ovariectomy (OVX). Iliac crest biopsies were separately processed before ovariectomy and at 60 days, 120 days and 180 days after ovariectomy. The histomorphometric changes were observed. It was found that the bone structure in trabecular bone remained relatively constant in the control group throughout the whole study. In contrast, the trabecular quantity was slightly decreased and the trabeculae were disconnected in OVX at 60 days after ovariectomy. The bone structure was characterized by the thinning of the trabeculae, the further discontinuance and quantity loss of trabeculae, and the anlargement of marrow cavity at 120 days to 180 days after ovariectomy. Apparently, these bone structural changes manifested with typical osteopenia pathological changes. Our results may serve as a basis for the design of further studies using OVX goat as an animal model for postmenopausal osteoporosis. PMID- 10683957 TI - [Circadian variations of plasma SOD and MDA in health subjects]. AB - Oxygen derived free radical system has been to be implicated in the pathogenesis of many diseases, such as cardiovascular disease, tumor, trauma and radiation injury. Many studies have suggested that oxygen free radical initiate a series of events that result in cell membrane alterations leading to the development of cell injury. The objective of the present study was to examine the circadian characteristics of some parameters of this system in the blood. One parameter was superoxide dismutase (SOD), a major oxygen free radical scavenger; the other was malondialdehyde (MDA), an important product of lipid peroxidation initiated by oxygen free radicals. Blood samples were collected at 6 h intervals beginning at 08:00 over a 24 h span in nine healthy volunteers (5 females, 4 males, average age 30 +/- 5 years). All participants were synchronized for one week with diurnal activity from 07:00 to 23:00 and nocturnal rest, with three meals served at 07:30, 12:00 and 19:00. Plasma SOD, MDA levels were measured by pyrogallol autoxidation assay, thiobarbituric acid spectrophotometry respectively, and the data were fitted with a 24 h cosine curve and analyzed further by a population mean cosinor. The results showed that the plasma SOD levels exhibited a highly significant circadian rhythm (P < 0.001) in healthy subjects, with acrophase (phi) at 21:17, mesor (M) of 3975.3 +/- 680.5 u/g Hb, and amplitude (A) of 273.62 u/g Hb. A circadian rhythm of plasma MDA was also detected (P < 0.01), phi = 15:15, M = 4.23 +/- 0.86 mumol/L, A = 2.37 mumol/L). The evaluation of circadian natures of SOD and MDA may be important in the prevention, diagnosis and treatment of associated diseases. PMID- 10683958 TI - [A study on P16 and P53 protein expressions in ovarian serous cystadenocarcinoma]. AB - An immunohistochemical method utilizing avidin-biontin complex (ABC) technique was used in this study to detect P16 and P53 protein expressions in 40 ovarian serous cystadenocarcinomas and 10 serous cystadenomas. The results showed that the total positive rates of P16 and P53 protein were 40% and 60% respectively. The positive staining rate was higher in P53 protein expression than in P16 protein expression in the same cases. The positive staining rates of P16 protein were 16.67%, 33.33% and 52.63% respectively in stages I, II and III. There was no significant difference in histological grading. The postive staining rate of P53 protein expression in poorly differentiated tumor was higher than that in well differentiated group (P < 0.05). These results suggested that P16 and P53 protein expressions of the ovarian serous cystadenocarcinoma might be correlated with human organs, histological type of tumor, cyclin and cyclindependent kinase. PMID- 10683960 TI - [Observation of the serum acidic isoferritin levels in patients with hepatocellular carcinoma]. AB - The levels of serum acidic isoferritin (SAIF) in 48 patients with hepatocellular carcinoma (HCC), 30 patients with hepatitis, 28 patients with liver cirrihosis, and 33 healthy subjects were measured by using enzyme-linked immunosorbent assay (ELISA--double-determinant). The result revealed that the SAIF values of HCC, HP and LC did not show normal distribution. The median values were 440 micrograms/L, 21 micrograms/L, 120 micrograms/L and, in normal subjects, 66 micrograms/L respectively. There were statistically significant differences between HCC and the control groups. The sensitivity of SAIF to HCC was 85.46%, with the cut-off point being 250 micrograms/L. SAIF was not correlated to AFP in HCC cases. The values of SAIF had no relationship with the volume of the tumors and the clinical stages. SAIF may be an available and useful serum marker and be beneficial to the diagnosis of early-staged HCC. PMID- 10683959 TI - [P53 protein expression in malignant germ cell tumor of ovary and its relationship with clinical course and prognosis]. AB - To study the relationship of P53 protein expression with clinical course and prognosis in malignant germ cell tumor of ovary P53 protein expression was examined by immunohistochemical ABC methods in 82 cases of the neoplasm and 20 cases of normal ovary tissue. The results showed that no expression of P53 protein was detected in normal ovarian tissue. The total expression rate of P53 protein was 24.39% in neoplasm, and the expression was not significantly correlated with the different histological types of neoplasm. The study however found that the protein expression of p53 gene was significantly correlated with the clinical stages and prognosis of the neoplasm. PMID- 10683961 TI - [Analysis of the mechanism on the formation of configuration of F-V curves with tangent time constant in adult humans]. AB - The tangent time constants at high, mid, low lung volumes (tau t75, tau t50, tau t25) were measured from F-V curves of 258 normal adults aged 20-49. The configurations of F-V curves were classified into the plateau, linear, convex and concave types. In terms of distribution ratio, the convex type increased with age, but the concave type decreased with age. The top of F-V curve was chiefly determined by the length of magnitude of tau t75. The magnitude of tau t75 was the longest for the concave type; its top fell down slowly with the formation of a blant round shape. The magnitude of tau t75 of convex type was the shortest; its top fell down rapidly and formed a peak shape. The mid portion of the curve was determined by the length of tau t50 and the height of V50. The tau t50 was longer and V50 was higher in the plateau and concave types, so that the mid portion fell down slowly to form a plateau or to be concave to the volume axis. While tau t50 was shorter and V50 was lower in the convex and linear types, the mid portion became convex to the volume axis or formed a linear shape. The configuration of the tail was determined by tau t25 and V25. The long tau t25 and high V25 in the plateou type implied the extension of plateau to the low lung volumes. While tau t25 in theother 3 types was shorter. However, V25 was higher in the concave type, thus it made the tail fall down fast and become concave to the volume axis. The V25 was lower in the convex type with the tail convex to the volume axis. The height of V25 in the linear type was between the concave and convex ones with the tail falling in a linear fashion. PMID- 10683962 TI - [Measurement and clinical significance of inspiratory drive efficacy in patients with chronic obstructive pulmonary disease]. AB - To study the relationship between VT/P0.1 and the function of inspiratory muscles, we performed the measurement of VT/P0.1 (inspiratory drive efficacy), MIP (maximal inspiratory mouth pressure) and MMIF (maximal midinspiratory flow) in 15 normal subjects and 60 patients with chronic obstructive pulmonary disease(COPD). The results indicated that VT/P0.1 decreased with the deterioration of COPD and it correlated highly and positively with MIP and MMIF. These suggest that VT/P0.1 can reflect the function of inspiratory muscles as well as MIP and MMIF can. PMID- 10683963 TI - [The value of erythromycin pleurodesis in the treatment of malignant pleural effusions]. AB - Malignant pleural effusion is a frequent complication of patients with advanced malignant tumors. For many patients suffering from malignant pleural effusion that increases rapidly and may be life threatening, it is important to control the effusions. To investigate the value of erythromycin as a pleural sclerosant, a dose of 1 g erythromycin in 30 ml of 5% glucose was injected into the pleural cavity of 26 patients with malignant pleural effusion. The results were assessed by Miller's standards. After treatment, numerous adhesions were present in 15 cases; Pleural effusion reduced in 7 patients, and no effect was noted in 4 patients. The total response rate was 84.6%, and there were no severe side effects. The erythromycin-induced pleurodesis is probably the result of induced chemical inflammation in the locality. This study suggests that erythromycin is a useful pleural sclerosant. PMID- 10683964 TI - [Observation on the changes of lipid peroxidation in neonates with sclerema]. AB - Thirty-six hospitalized newborn infants with sclerema (case group), which were divided into mild group(n = 18) and severe group(n = 18), and 28 normal neonates (normal control group) were selected to measure the changes of blood lipid peroxidation (LPO) and superoxide dismutase(SOD) so as to study the role of free radicals and lipid peroxide injury the pathogenesis of the entity. The results showed that the plasma LPO contents were significantly increased and SOD were significantly decreased in the case group when compared with those in the normal group (P < 0.01). The plasma LPO was significantly higher and SOD activity was significantly lower in the severe group than those in the mild group (P < 0.01). The study indicates that the free radicals produced in sclerema neonatorum are great in amount and suggests that free radicals possibly participate in the pathogenesis of the disease. PMID- 10683965 TI - [Analysis and prevention of road traffic fatalities in Sichuan Province]. AB - This is an epidemiological survey of the fatalities from road traffic accidents in Sichuan. All fatal accidents that occurred in Sichuan from January 1994 to December 1994 were analysed. The mortalities per million kilometers of vehicle travel, per 100,000 registered motor vehicles and per 100,000 resident population were 4.35, 42.41 and 3.92 respectively. These figures were compared with international death rates and those reported by other countries. The results showed that Sichuan had higher mortalities per million kilometers of vehicle travel and per 100,000 registered vehicles but lower mortality per 100,000 resident population, compared with some industrialized and rich develeping countries. The majority of the victims were males aged 21 to 40. Road traffic fatalities constituted the leading cause of all accident deaths. The cost of road accidents in Sichuan was estimated to be sixty million Yuan. Further strategies and methods to minimize the rate of such accidents have been suggested. PMID- 10683966 TI - [Improved MTT colorimetric assay for serum TNF activity]. AB - The activity of serum tumor necrosis factor(TNF) is known to be related with the mechanism and prognosis of many diseases. The aim of this experiment was to look for an economic and reliable method for assaying the activity of serum TNF. We used the neutrophil solventor 20% SDS-50% DMF instead of the acidity solventor 0.04 mol/l acidified isopropy alcohol and established an improved 3-(4,5-dimethyl thiazoly)2,5-diphenyl-tetrazolium bromide (MTT) colorimetric assay for measuring the amount and function of living cell. We used this improved MTT colorimetric assay for measuring TNF activity of peripheral blood serum of healthy persons. It avoided the deposition of the protein in serum and medium and showed more repeatability, as compared with the conventional MTT colorimetric assay. The results showed: when the activity of target cell(TC)-L929 is > or = 95%, the density of TC is > or = 1 x 10(2)/well and the number of living cells in each well is positively correlated with the OD volume (OD570nm-OD630nm of purple formazan metabolite of MTT solution(r = 0.87, P < 0.01); when the density of TC is 5 x 10(4)/well, the level standard of TNF in each well is negatively correlated with the OD volume (OD570nm-OD630nm)r = 0.79, P < 0.01). Using this method, we measured the TNF activity of 20 healthy persons' peripheral blood serum. The mean +/- s of TNF activity is 20.95 +/- 3.2 IU/ml. This method is dependable, easy-to-do and economic, it has good repeatability within 3-12 hours. PMID- 10683967 TI - [Preparation of chromosomal DNA from whole blood without use of phenol]. AB - We describe in this paper a simple and efficient procedure for preparing peripheral blood DNA without pre-isolation of white blood cells. The method also avoids the hazard of organic solvent extraction. Routinely it yields 20 to 28 micrograms of chromosomal DNA from per ml whole blood. The A260/A280 ratio for DNA sample is 1.80 to 1.85. The undigested DNAs migrate above the 21 kb pair, while the endonucleasedigested samples appear characteristically in the smear form. These data illustrate the consistency and quality of the chromosomal DNA isolated using this procedure. The DNA prepared by this method was used for analysis of beta-thalassemia mutation and prenatal diagnosis in our laboratory. PMID- 10683968 TI - [Quantitative determination of dracorhodin in Daemonorops draco B1. and traditional Chinese medicines containing Daemonorops draco B1. by HPLC]. AB - A reversed phase high performance liquid chromatographic mertod for the determination of Dracorhodin in Daemonorops draco B1. and Traditional Chinese Medicines containing Daemonorops draco B1. has been developed so as to set the quality standard for Sichuan Jinyao. The result showed that the components of Dracorhodin were separated by ODS column, with MeOH-0.05 mol/L NaH2PO4 (51:50) (pH = 3.10-3.15, adjusted with H2PO4) acting as the mobile phase. The other high retentive components were eluted by methanol; thus the analytical time was reduced. The method is simple and accurate. PMID- 10683969 TI - [Cloning and expression of leptospiral protective antigen gene OmpL1 in BCG]. AB - This study was intended to produce a new living vaccine against leptospirosis using BCG as vector. Leptospiral outer envelop antigen gene OmpL1 was amplified from the genome of pathogenic leptopira serova Lai 017 by PCR, and cloned in E. coli-BCG shuttle plasmid pY6002. Recombinant plasmids were isolated by dot blotting with Digoxigeninlabeled OmpL1 gene. After transforming the recombinant plasmids in BCG (Shanghai strain) by electroporation, the genomic DNA of all 21 transformants were prepared and hybridized with OmpL1. It showed that 6 of the 21 transformants were recombinants in which the OmpL1 gene had been integrated into the genome of BCG. By immunoblotting with OmpL1 infected rabbit antiserum, which was preabsorbed to remove antibody against E. coli and SPA-HRP, three recombinants, pLI1, pLI2 and pLI3, were detected to express OmpL1 protein. The ability of expression is in the order of pLI2 > pLI1 >> plI3. These studies provide the possibility of further research on the development of highly efficient recombinant vaccines against leptospirosis. PMID- 10683970 TI - [Relationship between the expression level of alpha-MHC gene and cardiac contractility during heart failure]. AB - To investigate the molecular basis of the decrease in myocardial contractility during heart failure, an animan model of heart failure was set up by means of deoxycorticosterone-acetate impregnated silicone rubber implants in wistar rats. Cardiac contractility in normal and heart failure rats was examined, and gene expression of its myocardial contractile protein, alpha-MHC, was quantitatively analyzed at gene transcription level by using RNA slot blot hybridization. The results showed that the cardiac contractility and the alpha-MHC mRNA levels in heart failure rats were all lower than those in the normal. Statistical analysis showed a positive correlation between the cardiac contractility and the gene expression of alpha-MHC (r = 0.4143, n = 43, P < 0.05). The above results indicate that gene expression level of alpha-MHC is one of the key factors determining cardiac contractility. PMID- 10683971 TI - [Study on the beta-lactamases of the Escherichia coli HX88108 resistant to ceforperazon]. AB - E. coli HX88108 was isolated from a patient and found to produce plasmid-encode beta-lactamases with conferring highly resistance to ceforperazone(CPZ). The beta lactamases of the E. coli HX88108 and transformants pFC, pFT1, pFT2 and pFT3 were studied. The beta-lactamases stability test among 11 beta-lactam antibiotics showed that beta-lactamases from E. coli HX88108. pFC, pFT1, readily hydrolyzed penicillins, the first, second-generation cephalosporins and CPZ. beta-lactamases of pFT2 and pFT3 hydrolyzed penicillins more strongly than cephalosporins. On the other hand, experiments of inhibiting enzyme were carried out. The results indicated that beta-lactamases of HX88108, pTF1, pFT2 and pFT3 were inhibited by clavulanic acid(CA) and sulbactam (SBT). Enzyme of pFC was inhibited poorly by CA and SBT. Through isoelectric focusing technique, the PIs were as follows: HX88108 contained three beta-lactamases, of which the PIs were 5.25, 5.3 and 5.6 respectively; the PIs of beta-lactamases from pFT2, pFT3, were 5.3 and 5.6. pFC and pFT1 were different plasmids encoded beta-lactamases with the same PI 5.25. The results indicate that the beta-lactamases of E. coli HX88108 may be a new member in TEM farmily. PMID- 10683972 TI - [Reversal of cancer multidrug resistance by Chinese medicine Ams-11, Fw-13 and Tul-17]. AB - The aim of this project was to find some kinds of Chinese materia medica as effective agents for in vitro reversal of cancer multidrug resistance. Based on the present authors' previous researches, thirty-two kinds of Chinese medicine as research meterials were selected and examined. Using cell growth inhibit assay, the authors found that three of them--Ams-11, Fw-13 and Tul-17 in the doses free from cytotoxity could enhance the sensitivity of multidrug resistant cells to anti-cancer drugs in a dose-dependent way. It seemed that these three kinds of Chinese medicine might be potential effective reversal agents. PMID- 10683973 TI - [Study on telomeric association in nasopharyngeal carcinoma]. AB - The chromosomal telomeric association in peripheral blood lymphocytes from 25 untreated patients with nasopharyngeal carcinoma (NPC) and in NPC cell lines (CNE) was investigated by using chromosome G banding technique. The cell telomeric association rate of patients (35.07%) was found to be higher than that of controls(20.27%) (P < 0.01), while in the two groups the distributions of chromosomes involved were approximately the same (P > 0.05). The telomeric association rates of patients were significantly higher than those of control in chromosomes 1, 2, 3, 4, 7, 11, 15, and 17(P < 0.05). In NPC lines (CNE), 94% of the cell division phases showed telomeric association. Chromosomes 2, 3, 5, 11 and 16 had higher frequencies of association than other chromosomes. The most frequent telomeric association was single chromatid association. PMID- 10683974 TI - [Isolation of antibacterial polypeptides of human cervical mucus]. AB - The acid-soluble extract of human cervical mucus was obtained by solving mucus with 5% acetic acid in the presence of protease inhibitor. The antibacterial activity of the acid-soluble extract was analyzed by gel overlay technique. The result showed that two protein bands which were designated human cervical protein 1 (Hcp-1), human cervical protein-2(Hcp-2) were potently antibacterial against E. coli 25922 and S. aureus 25923. Tricine-SDS-PAGE analysis indicated that Hcp-1, Hcp-2 actually contained three and four protein bands respectively. The molecular weights of Hcp-1 were 6.7 kd, 10 kd, 15.4 kd; these of Hcp-2 were 4.4 kd, 6.7 kd, 9 kd, 15.4 kd. Our studies suggested that human cervical mucosa might secrete some currently-unknown antibacterial polypeptides which play an important role in the cervical defense against infection. PMID- 10683975 TI - [Distribution of morphine in acute morphine-treated rats]. AB - The distribution of morphine in acute morphine-treated rats was studied by immunohistochemistry (PAP). Samples were examined for morphine distribution and localization in internal organs at 10-160 min after intraperitoneal injection of morphine. The results showed that morphine was distributed in the heart, hepatocytes, epithelial cells of the bile canalicular cells in liver, spleen, kidney, adrenal glands, stomach parietal cells, small intestinal villus, pancreas islet, sublingual glands and submandibular glands. Morphine was found in the cardiac tissue, liver, spleen, pancreas islet, small intestinal villus at 10 min after injection and in the cardiac tissue, stomach parietal cells at 160 min after injection. The cause of the postmortem the distribution of morphine in internal organs at different times after injection was discussed. The results of this study may be used as a basis for the choice of samples for the forensic toxicological analysis. PMID- 10683976 TI - [The effect of removable partial denture on the composition of bacteria on the apt-to-decay site of abutment teeth]. AB - This study was aimed at the composition of plaques which were on the base teeth's carious sensitive sites, just after the insertion of the removable partial denture (RPD). The results showed that the ratios of bacterial detection in different teeth at the same period and those on the different sites of same teeth at different periods were not significantly different. There was a tendency that the detection ratio of the stomatococcus at the early period was higher than that of other bacteria, but at the middle period the ratios declined, and at the late period it was higher again. The detection ratio of the bacillus was on the contrary in the study. At the same time, the composition of the plaques changed significantly after the dentures had been used for 7, 14, and 21 days and 3 months. After 3 months, the ratios of the stomatococcus and the bacillas returned to their normal levels. The 3 main cariogenic bacteria were all detected in this study. The Streptococcus mutant was the dominant bacterium in the plaques and its ratio went up with the time duration. The ratio for the lower second molar was higher than that for the mandibular second premolar (P < 0.05). The detection ratios of the Actinomyces viscosus and lactobacillus were lower. In conclusion, after the insertion of RPD, the oral microbial ecosystem would be changed and the cariogenic bacteria began to implant on the surface of the base teeth. So RPD is a potential factor to cause the caries. PMID- 10683977 TI - [Expression of CD44 molecule and its significance in uveal and conjunctival melanomas]. AB - The aim of this study was to evaluate the role of over expression of CD44 molecule in the development and progression of uveal and conjunctival melanomas. Flow cytometry (FCM) and immunofluorescence methods were used for detecting the CD44V expression in uveal malignant melanomas (UMM), conjunctival nevi (CN) and conjunctival malignant melanomas(CMM). The expression content of CD44 in 7 cases of CMM was significantly higher than that in 5 cases of CN (P < 0.05); the CD44V positive expression percentages in 7 cases of CMM and 40 cases of UMM were 71.43% and 62.50% respectively; the expression content of CD44V in UMM was related to scleral invasion (P = 0.0105); there was a negative correlation between the expression content of CD44V and proliferative index (PI), S-phase fraction (SPF) (P < 0.01; P < 0.05). The results suggested that the over expression of CD44V might be involved in the development of CMM and UMM and related to local infiltration ability of UMM and that the CD44V expression content detected by FCM might be helpful in discriminating CN From CMM, but this waited for further research confirmation. PMID- 10683978 TI - [The LDL receptor activity of hepatocytes during cholesterol gallstone formation in rabbits]. AB - In order to study the mechanism of cholesterol gallstone formation through rabbit model which was induced by high cholesterol diet, we investigated the LDL receptor activity of hepatocytes binding to 125I-LDL in different phases, namely 1, 2, 3 week group and 4 week, and in a control group besides. In this animal experiment, cholesterol gallstones were induced at 2 week, 3 week and 4 week groups in 4/10, 6/10, and 7/10 cases respectively. The Bmax values of LDL receptor of hepatocytes binding to 125I-LDL decreased significantly in 3 week and 4 week groups (vs 1 week and control groups, P < 0.05). The kd values became increased in 3 week and 4 week groups (vs 1 week and control group, P < 0.05), which suggested that the activity of LDL receptor decreased gradually. In conclusions owing to the intake of high cholesterol diet with the passage of time, the decreased activity of LDL receptor of hepatocytes would reduce the synthesis of bile acid. PMID- 10683979 TI - [Changes of cerebral blood flow during hypoxicischemic encephalopathy in newborn pigs]. AB - This study was designed to explore the change of cerebral blood flow during hypoxic-ischemic encephalopathy (HIE). Sixteen newborn pigs were devided into four groups: the normal control, 15 minutes after HIE, 30 minutes after HIE and 60 minutes after HIE. The shapes of the cerebral blood vessels were obsessed by the prepared Chinese ink irrigation and the width and volume fraction of capillary were tested and analyzed by stereographic method respectively. The results showed the small blood vessels were interrupted with the passage of time. At the time of 60 minutes after HIE, capilaries almost disappeared from view, only a few ends of blood vessels in the sheap of a broken tree were seen. The width of capillaries and small veins became more and more narrow as time went on. The percentages of capillary volume fraction and small vein volume fraction became smaller and smaller (P < 0.05). These demonstrate that the fall of cerebral blood flow plays an important role in the development of HIE. PMID- 10683980 TI - [Mechanism for the formation of F-V curve configuration analysed with tangent time constant and wave-speed theory in normal adolescents]. AB - To understand the distribution of F-V curve configuration and the characteristics of tangent time constant (tau t), and to explore the mechanical mechanism for the formation of F-V curve configuration, we measured the tau t of high, mid, low lung volumes (tau t75, tau t50, tau t25) from F-V curves in 728 normal adolescents aged 8-19. The results showed that the distribution of the concave type was the highest among 4 types of F-V curve configuration with a range of 40% 70%. Its tau t features were tau t75 > tau t50 > tau t25 and magnitude of tau t75 > 1s. Then the concave type and its tau t features were called "adolescent fashion". Contrastedly, the distribution of the convex type was the lowest, only 4%-22%. The mechanical mechanism responsible for the formation of the concave type was analysed with the wave-speed theory. During the growth of adolescents, the lung elastic pressure and driving pressure increase with age. The diameters of airways increase, especially those of the peripheral airways with profuse branches. These changes lead to a decrease of resistance. Therefore, the critical flow (Vc) or wave-speed flow (Vmax) of peripheral airways is raised, and the flow there-in could hardly be limited. Thus the choke point (CP) will stay in trachea for a longer period, leading to the elongation of the plateau towards the tail or the concavity of the curve towards the volume axis and hence the formation of the concave type. PMID- 10683981 TI - [A study of the adenosine deaminase polymorphism in a Chinese Han population by the PCR method]. AB - The study of genetic polymorphism of adenosine deaminase (ADA) has not been reported in China and abroad. The target DNA was amplified by PCR. The amplified product was then digested with Pst I, and run on a 9% polyacrylamide gel electrophoresis and followed by AgNO3 staining. ADA polymorphism, was determined in 80 unrelated individuals Chinese Han polulation. Five alleles and 14 genotypes were detected. The gene frequencies were ADA * 10 = 0.15625, ADA * 11 = 0.31875, ADA * 12 = 0.23750, ADA * 13 = 0.23125 and ADA * 14 = 0.05625 respectively. The heterozygosity was 0.8 and the discrimination power (DP) was 0.89. The distribution of observed genotypes frequencies was in good agreement with the Hardy-Weinberg law. The study of three families demonstrated that this locus was inherited according to Mendelian law. The technique is quite simple, fast, extremely sensitive, and it does not require the use of radioisotopes. This new method may be used for the forensic identification and paternity testing. PMID- 10683982 TI - [Determination of copper, iron and zinc in hair of mentally retarded children by oscillopolarography]. AB - The oscillopolarographic method (proposed by Ding Jianwen)for the simulltaneous determination of copper, iron and zinc in hair was further studied and applied to the investigation of copper, iron, zinc content in hair samples from 46 mentally retarded children and 49 healthy children. For copper, iron, zinc, the detection limits were 0.03, 0.03, 0.01 microgram/ml, dynamic range 0.05-15, 0.05-2.5, 0.2 10 micrograms/ml, the RSD of sample analysis 3.2%, 3.3%, 2.4%, the recoveries 96: 0%-105.2%, 2%, 97.3%-109.0%, 94.7%-107.7%, respectively. The difference between the results for the same sample by this method and the atomic absorption spectroscopic method was no statistical significance. The results showed that the copper content and iron content of hair samples from mentally retarded children were significantly lower than those of healthy children. PMID- 10683983 TI - [The risk of lung cancer and mesothelioma in farmers exposed to crocidolite in environment]. AB - To assess the risk of lung cancer and mesothelioma after environmental exposure to crocidolite for 20-30 years, a retrospective cohort study was carried out in farmers who had been exposed to crocidolite in environment. 1610 subjects were followed up for 9 years (Jan. 1, 1987 Dec. 31, 1995). The control group consisted of 7646 farmers who resided in the noncrocidolite pollution rural area in the same province. The results showed that the annual mortality rate was 43.75 per 100,000 population for lung cancer, and 36.46 per 100,000 for mesothelioma. Significantly high risks of lung cancer (RR 5.67) and mesothelioma (RR 182.3) were noted. These results demonstrate a strong causal association between lung cancer, mesothelioma and exposure to crocidolite. PMID- 10683984 TI - [Study on the power of O/E method]. AB - This paper addresses the methodology of calculating the power of O/E method and introduces the relevant algorithm for estimating the sample size. Based on the principles of hyporthesis testing and Poisson distribution, the power can be worked out for given sample size, baseline rate(pi 0) and the factor of increasing (gamma). The power increases with the increase in sample size, pi 0 and gamma. In shape the power is not a continuous smooth line but a zigzag line. The method illustrated in this paper can be also used for estimaging the sample size of studies on the intervention of diseases. PMID- 10683985 TI - [Pathologic grading and differential diagnosis on oligodendroglioma]. AB - To investigate the histological characteristics of oligodendroglioma (ODG), 65 cases of ODG were observed with light microscopy and partially with immunohistochemistry. According to WHO classification of tumors, 65 cases of ODG were graded in degree of differentiation of tumor cells. Of these, 19 cases belonged to grade I, 32 grade II, and 14 grade III. CONCLUSION: Except the degree of differentiation, some other criterlons as cellular density, mitotic activities, and the presence of focally concentrated apoptotic cells are closely related to grading of ODG. PMID- 10683986 TI - [Cytokines release related to cardiopulmonary bypass in patients with prosthetic valve replacement]. AB - To examine cytokines release related to cardiopulmonary bypass(CPB), the anthers investigated the possible differences in cytokines responses between patients undergoing prosthetic valve replacement (study group, n = 10) and those receiving closure of patent ductus arteriosus or pericardiectomy(control group, n = 9). Venous levels of interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interleukin-10 (IL-10) were measured at multiple time points before, during and after operation. As compared with pre-operative values, IL-2 levels in both groups decreased significantly (P < 0.05), and the levels of sIL-2R, IL-6, TNF-alpha, and IL-10 in both groups increased significantly at multiple time points post-operative (P < 0.01). All the values of cytokines returned to pre-operative levels at 7th post operative day. Although there were no pre-operative differences in these cytokines between the two groups (P > 0.05), the post-operative changes of cytokines in study group was more obvious than that in control group (P < 0.01). In study group IL-10 rose to a peak value of around 620 pg/ml at the end of CPB while IL-6 and TNF-alpha levels reached their peak values of around 88 pg/ml and 52 pg/ml respectively at 1st post-operative day. IL-10 has been reported as an antiinflammatory cytokine. The preceding IL-10 peak value, as compared with the peak values of IL-6 and TNF-alpha, could be associated with the interplay and regulation of cytokine network. On the other hand, in control group the levels of IL-6, TNF-alpha, and IL-10 reached their peak values at 1st post-operative day simultaneously. The values were 34 pg/ml, 36 pg/ml, and 162 pg/ml respectively. This result suggests that besides surgical stress mediated cytokines production or suppression, CPB itself also results in obvious changes of cytokine metabolism. However further studies are needed to elucidate the underlying mechanisms and clinical value of post-operative cytokines production or suppression related to CPB in patients with prosthetic valve replacement. PMID- 10683987 TI - [Cromakalim inhibits endothelin-1 induced pulmonary hypertension in rats]. AB - There is evidence that endothelin-1 (ET-1) and potassium channel may play an important role in the development of pulmonary hypertension. To evaluate the effect of ATP-sensitive K+ channel opener on pulmonary hypertension induced by ET 1, catheter was inserted into the pulmonary artery in ten male Wistar rats which had had pulmonary hypertension established by infusion of ET-1 (1.5 micrograms/kg), and then cromakalim were injected with a dose of 150 micrograms/kg. The mean pulmonary arterial pressure (mPAP), cardiac output (CO) monitored before and after infusion of ET-1, and 1 min, 5 min, 10 min after cromakalim injection, and pulmonary vascular resistance (PVR) were calculated. It was found that the mPAP was significantly increased, from 2.36 +/- 0.24 kPa to 3.32 +/- 0.49 kPa(P < 0.01), and PVR also increased, by infusion of ET-1. After cromakalim injection, mPAP were decreased to 2.50 +/- 0.62 kPa in 1 min, 1.14 +/- 0.18 kPa in 5 min and 2.33 +/- 0.52 kPa in 10 min, PVR decreased significantly. It is suggested that there is interaction between ET-1 and potassium Channel, and Cromakalim decreases mPAP in part by inhibiting the response of pulmonary artery to ET-1. PMID- 10683988 TI - [Semiquantitative measurement of progesterone receptors in luteal-phase-defect endometrial cells during secretory phase]. AB - To investigate the changes of endometrial progesterone receptor (PR) of luteal phase-defect (LPD) patients during the secretory phase, thirteen patients with complaints of infertility or habitual abortion were studied. During the early-mid secretory phase, endometrial tissue was obtained by dilatation and curettage (D & C) for histological and receptor study: meanwhile serum E2, P, FSH, LH and PRL were measured. Based on histologic diagnosis, the patients were divided into two groups: the LPD group (n = 7) and the normal control group(n = 6). PR content was determined by immunohisto-chemical (IHC) assay. The results showed that during the early-mid luteal phase a significantly low PR content on endometrial glandular nucleus was observed in LPD group, compared with normal control(6.75 +/ 2.57 vs 9.50 +/- 1.64 P < 0.05), but no difference in serum progesterone was noted between the two groups. These findings suggest that during early-mid secretory phase, PR content on endometrial glandular nucleus decreases in LPD cases, which results in deficient response of endometrium to proper stimulus of progesterone. This change may cause endometrial secretory deficiency and blockade of embreyo implantation. That is why infertility or habitual abortion happened. PMID- 10683989 TI - [Comparison of the effects of inhaled nitric oxide and intravasculare regitine on pulmonary gas exchange in young dogs with oleic-acid acute lung injury]. AB - In order to find an agent which truly improves hypoxemia of some serious pediatric lung diseases, the authors examined the independent effects of nitric oxide inhalation and regitine infusion on blood gases, intrapulmonary shunt and hemodynamics in young dogs with oleic-acid acute lung injury. After nitric oxide inhalation, the results showed moderate increases in PaO2 and SaO2 (P > 0.05) and a significant decrease in Qs/Q tau ratio (P < 0.01). There was a significant decrease of PAP(P < 0.05), while SAP remained unchanged. After regitine infusion, however, there were marked decreases in PaO2 and SaO2 (P < 0.01); meanwhile, Qs/Q tau rose (P < 0.05). These suggest that with the presence of pulmonary pathology nitric oxide inhalation may alleviate the elevated pulmonary pressure without alteration in systemic artery pressure; so it can improve pulmonary ventilation perfusion distribution and cause favorable changes in blood gases. On the other hand, regitine, as a non-selective vasodilator, reduces pulmonary artery pressure at the cost of significant worsening of blood oxygenation and systemic hypotension; so its routine use in childhood pulmonary diseases should be cautiously considered. PMID- 10683990 TI - [The relation between nutrition and pulmonary ventilation in patients with chronic obstructive pulmonary disease during the relieved period]. AB - To observe the effect of nutrition on pulmonary ventilation in aged patients with chronic obstructive pulmonary diseace (COPD) during the relieved period, the pulmonary function (MVV, PVC, FEV1, MMEF) of 261 aged patients with COPD was tested during the relieved period. The results showed that the MVV (48.75% +/- 19.9%) tested in the patients with malnutrition was significantly lower than that tested in the patients with normal nutrition (59.1% +/- 22.6%) (P < 0.01) while there was no marked difference in the ventilation in their large and small airway. It is conluded that malnutrition of the patients with COPD affects the constrictive function of respiratory muscle and thus results in a decrease in MVV. PMID- 10683991 TI - [Esophageal reconstruction using ilecolon in children--a report of ten cases]. AB - The use of the ileocolon as a substitute for esophagus in children has been a well-established surgical procedure. Between 1992 and 1995, ten retrosternal esophageal substitutions using an ileocolic interposition were performed. The ages of the children ranged from 2 to 8 years. The esophageal strictures were secondary to ingestion of caustic liquid. No death occurred in the intra- and post-operation period. Cervical ileoesophageal anastomotic leaks took place in 2 cases as the early major postoperative complications, which healed spontaneously. Three cases of ileoesophageal anastomotic strictures were noted in the follow-up period. The surgical indications, the treatments of anastomotic leaks and strictures, and the importance of long-term follow-up were discussed. PMID- 10683992 TI - [Free jejunal grafts for reconstruction of pharynx and cervical esophasgus]. AB - Eleven patients with hypopharyngeal carcinoma were treated surgically and the pharynx and cervical esophagus were reconstructed with free jejunal grafts through the technique of microvascular anastomosis. The microvascular anastomosis were all successful and the patients all survived the operations. The operations were performed at one time with only a few complications. There were 2 cases of pharyngeal fistula and 1 case of bleeding from the anastomotic site. The patients could eat by their mouths and swallow very well just 15 days after the operation. The 3-year suvival rate was 36%. The main causes of death were the recurrence and the metastasis to the cervical lymph nodes. We came to the conclusion that the operation is safe with high success rate and no mortality during operation, and it is less injurious than the gastropharyngeal anastomosis. The key point of the operation is the surgeon's mastery of the microvascular anastomosis technique. PMID- 10683993 TI - [A retrospective study of neurogenic pulmonary edema following intracranial injury without open wound]. AB - To find factors that relate to the occurrence of neurogenic pulmonary edema(NPE) following intracranial injury without open wound, thirty-founr autopsies on such cases were studied retrospectively. The rate of NPE in the 34 cases was 41.18%, and it was 44.44% in those aged 10 to 49. The rate of NPE in the persons who died immediately or 0-6 hours after injury was 41.18%, and in those who died 7-48 hours after injury, 41.18% too. The rate of NPE was 69.23% for the cases with only one site of bleeding and 23.81% for ones with multiple sites of bleeding. These findings suggest that the occurrence of NPE is not related with the duration of time after head injury or the location of brain injury, but it is related with the severity of brain injury. PMID- 10683994 TI - [Isoelectric focusing and immunoblotting identification of human ApoE phenotype]. AB - A simple, rapid and accurate method for phenotyping sera apoE has been developed. In this method 10 microliters serum or plasma are incubated with Dithiothreitol and Tween-20 for 15 min and then applied to 5% polyacrylamid gel containing pH 4 8 ampholyte and 3 mol/L urea. After 2 h focusing, the apoE bands are transferred to Nitrocellulose membrane. apoE bands are made visible by immunoblotting and TMB is used as the substrate. Identification phenotype is easily accomplished by noting the location and number of protein bands. 56 samples shows apoE 3/3 in 47 cases, apoE 2/3 in 2 cases, and apoE 3/4 in 7 cases. This method is well suited for large-scale population studies and clinical labs. PMID- 10683995 TI - [Extraction and purification of mouse H-2 antigen]. AB - This study was designed to purify the mouse major histocompatibility complex antigen (H-2Ag). The detergent was used to extract the crude H-2Ag, and monoclonal antibodies affinity chromatography was applied to purify H-2 antigen. A 45 kd heavy chain and a 12 kd light chain from the purified proteins were shown by electrophoresis. Besides, the pure H-2 antigen was found to have immunological and biological activity. This method should be useful in purifying large quantities of H-2Ag for the researches in transplantation and functional analysis of H-2 antigen. PMID- 10683996 TI - [The effect of a vaccine made from 39kd hydrophobic outer membrane protein of Leptospira interrogans on neurohumoral and red cell immunity function of the guinea pigs]. AB - A randomized control trial was conducted to determine the immunoprotective efficacy of OmpL39. 36 guinea pigs were divided into OmpL39 group, whole leptospiral cell vaccine (WLCV) group, other proteins of Leptospira group, and negative control group (normal saline, NS). The results showed that all the guinea pigs of infected OmpL39 and WLCV still survived, but all the control guinea pigs died. Immunoprotective efficacy was 100% for OmpL39 and WLCV. OmpL39 levels were similar to WLCV levels and higher than controls (P < 0.05). These suggested that OmpL39 could be used as important immunoprotective antigen to develop the genetic vaccine of the targeting delivery system. Also it was observed that OmpL39 could produce 100% immunoprotective efficacy, have higher MAT level (> 1:3200) and regulate 5-HT, 5-HIAA, DA, NE. The 5-HT, DA, NE concentration was lower and 5-HIAA was higher than that of controls after stimulating the guinea pigs with OmpL39 (P < 0.05). The results suggested that OmpL39 genetic vaccine could produce immunity function and have an active effect on absorbing inflammation and protecting organs and tissues. Besides, the red cell immunity efficacy of the guinea pigs was changed during immunity response and the RBC-C3b RR and the RBC-ICR were increased. The RFER was increased; the RFIR was decreased. RFER/RFIR was remarkably higher than that of control (P < 0.05). These suggested that OmpL39 could increase red cell immunity function. PMID- 10683997 TI - [Partially purified antibacterial polypeptides from granules of human large granular lymphocytes]. AB - This was a study on the antibacterial activity of human large granular lymphocytes (LGL). In our previous work, three antibacterial fractions named HLP 1, HLP-2 and HLP-3 had been identified from the acid-soluble extracts of the granules of LGL by using the gel overlay technique. The present study demonstrated that HLP-1, HLP-2 and HLP-3 separated by preparative acid-urea polyacrylamide gel electrophoresis were potently bactericidal against E. coli MI 35P, S. aureus ATCC 25923, P. aeruginosa ATCC 27853 when tested for their antibacteiral activities by using both agarose radial diffusion assay and gel overlay technique. SDS-PAGE analysis showed that the HLP-3 was almost purified with the molecular weight of 7 kd. The HLP-1 and HLP-2 were more complex, and the ranges of molecular weight were 5.6-13 kd and 5.6-8.8 kd respectively. Our study suggests that the granules of human LGL might contain some low molecular weight antibiotic peptides which play an important role in human innate immunity. PMID- 10683998 TI - [Cloning and orientation of cefoperazone resistance gene on plasmid pFC in E. coli HX88108]. AB - Cefoperazone resistance gene (CPZr) has been cloned from plasmid pFC of E. coli HX88108 using the vector pMB9 (TCr, 5.3 kb). The plasmid pFC DNA was partially digested with Sau3A I, and its 1-2 kb fragments were ligated into BamH I site of vector plasmid pMB9. The recombinant DNA was then transformed into E. coli DH5 prepared using calcium chloride. CPZ resistant bacterial colonies were selected on the agar SOB plates containing CPZ (40 micrograms/ml). The resistance to CPZ could be stably reserved in generation after generation. The recombinant plasmids which encoded CPZ resistance were designated pFL11, pFL25, pFL33, pFL82, pFL86 and pFL102. Rapid small-scale preparation of plasmid and DNA restrication enzyme analysis were used for identification of bacterial colonies. Five plasmids DNA physical maps have been established. Comparison of recombinant plasmids maps with pFC map confirmed that the CPZr gene was oriented between nucleotide no. 3200 bp and no. 4800 + 40 bp of plasmid pFC total sequence. Its molecular weight was about 1.6 kb. There were EcoR I, Sma I and Pvu II sites within CPZr gene. PMID- 10683999 TI - [Study of macroencapsulated islet xenografts for treatment of diabetes in mice]. AB - The aim of this study was to prevent rejection after islet xenotransplantation. Islet obtained from Wistar Furth rats were macroencapsulated with agarose and collagen before transplanted into diabetic C57BL/6, B6AF1, and BALB/c mice. The results showed that the diabetic condition was reversed in 92.3% of recipients transplanted with macroencapsulated islets. The normoglycemic state of recipients was maintained 125.8 +/- 57.9 days without immunosuppression. The glucose tolerance curves in these mice were similar to those of normal mice. The macroencapsulated islets retrieved 103 days after transplantation showed no evidence of tissue reaction or fibrosis. These indicated that the macroencapsulated islets serve both to achieve and maintain normoglycemia of recipients, as well as to protect islet xenografts from rejection. PMID- 10684000 TI - [Study of flow field uniformity downstream of mitral stenosis using Doppler echocardiography]. AB - To research into the relationship between mitral lesion, stenosed degree and flow field uniformity downstream of mitral valve, we adopted the advanced color Doppler echocardiographic technique to conduct a quantitative study of flow field uniformity downstream of mitral valve in sixty patients with varying-degree of mitral stenosis in vivo. Twenty normal persons acted for comparison. The results showed that there was a linear correlation between the extent of valvular lesion, the stenosed degree and flow field uniformity downstream of mitral valve. We conclude that the more severe the mitral lesion is, the more grave the stenosis is and the worse the flow field uniformity is. This suggests that there might be a reciprocity between valvular lesion and flow field uniformity as well as turbulent shear stress, which should be further studied. PMID- 10684001 TI - [A pharmacokinetic study of 125I-VIP in rat]. AB - To investigate the pharmacokinetics of 125I-VIP in normal rats, a single dose of 125I-VIP was given intravenoualy to rats and from 0.5 to 240 min after the administration, serial blood samples were taken and measuredp by gamma scintillation counter and finally expressed as mu Ci/ml blood. The data were analyzed by computer for the estimation of pharmacokinetic parameters and the judgment of compartment model with a program of 3P87. The results demonstrated that the pharmacokinetics of 125I-VIP in rat fitted with the three-compartment model(Wi = 1) based on either the comparison of the calculated theoretic value with measured concentration, the values of AIC and r2 or F test for the model judgment, and that the average distributive half-life of 125I-VIPk in blood was 1.36 min, the elimination half-life(T1/2 beta was 67.3 min, the K12(3.589 min-1) was markedly higher than that of K12(0.045 min-1), and the K21:K12(0.0127) was obviously lower than that of K31:K13(0.215). The present study indicates that 125I-VIP has a rapid uptake and slow elimination in the tissues with high density and affinity VIP receptors (the second compartment). PMID- 10684002 TI - [Study of a new type of material substituted for human hard tissues-hydroxyl poly calcium sodium phosphate: histologic report implants in mandible of animals]. AB - This study was aimed to evaluate the histocompatibility of hydroxyl poly calcium sodium phosphate (HPPA) preliminarily. HPPA and hydroxylapatite were implanted in the mandibular defect in dogs. The implant-bone interface was observed by using stereomicroscope and decalcified histologic sections. The result demonstrates that HPPA has well-osseous integrated property after being implanted into bone tissues. This suggests that HPPA might be a new material of future excellence in substitution for human hard tissues. PMID- 10684003 TI - [Effects of surface roughness of two restorative materials on early Streptococcus sanguis adhesion in vitro]. AB - This was an in vitro study aimed at the surface roughness of two kinds of commonly used restorative materials resin and alloy necessary to affect the adhesive behavior of Streptococcus sanguis(S. s) on them after 24 h. The surface roughness(SR) of each tested sample was evaluated with a profilometer quantitatively and observed with scanning electron microscope (SEM) morphologically. Then the adhesive microbial amount was determined by the clone forming unit counting method, and adhesion morphology was analyzed with SEM. The result showed a positive linear relation between the adhesion amount of S. s and the SR. The relative coefficients were rresin = 0.46(P < 0.01) and ralloy = 0.25(P < 0.01) respectively. These suggest the SR of the restorative material is important for the early adhesion of oral microbes in vitro. Therefore, before the prothesis is inserted in the patient's oral cavity, its surface should be polished as smoothly as possible so that the bacterial adhesive amount on its surface can be decreased, and hence the patient may keep in good oral health and have a prolonged use of the prothesis. PMID- 10684004 TI - [HA-coated titanium implants used as orthodontic anchorage. An experimental investigation of implant stability and peri-implant neck tissue in dogs]. AB - This study was aimed at the feasibility of using HA-coated titanium endosseous implants as orthodontic anchorage. These implants installed in dog's mandibles were loaded with the orthodontic force of 150 g for 3 months. The stability of the implant and the peri-implant neck tissue were investigated with radiograph and index evaluation. The findings demonstrated that no implant was mobile or loose or dislocated. The soft tissue around the cervical part of the implants had a slight inflammation because of bad oral hygiene and the stimulation of residual food attached to the stainless string. However, no resorption of marginal alveolor bone was found under sustained orthodontic force. The above results suggest that the HA-coated titianium implant can be used as anchorage for short term orthodontic treatment. PMID- 10684005 TI - [Protective effects of flunarizine and vitamin C on isolated rat heart subjected to ischemia-reperfusion injury]. AB - Isolated rat heart was subjected to Langendorff perfusion for 10 minutes of total global ischemia, and reperfusion for 15 minutes. The content of myocardial intracellular calcium, content of oxygen free radical induced lipid peroxide mitabolite-malondialdehyde (MDA), content of Lactic dehydrogenase (LDH) in coronary effluent, and the change of myocardial morphology were studied. The effects of flunarizine(FNZ) and/or vitamin C(Vit C) on the above parameters were observed. The results showed that, at 15 minutes of reperfusion, the contents of myocardial intracellular calcium, of MDA in myocaredium and of LDH in coronary effluent in the FNZ + Vit C group, Vit C group and FNZ group were all lower than those in the control group, and the change of morphology in the three groups was also slighter than that in the control group. When FNZ and Vit C were administered together, the effect was more marked and equivalent to the sum total of the effects of the two drugs. These suggest that both FNZ and Vit C reduce reperfusion injury through suppression of myocardium calcium overload and oxygen free radical damage. PMID- 10684006 TI - [Relationship between nitric oxide synthase, endothelin and the initiation of hypertension and atherosclerosis]. AB - To investigate the relationship between nitric oxide synthase (NOS), endothelin (ET) and the initiation of hypertension and atherosclerosis, 5 hypertensive rats (systemic pressure 22.7-24 kPa), 5 atherosclerotic rats (by cholesterol-rich diet, 3 months), and 5 normal Wistar rats were used for determination of the distribution of NOS and ET of aorta by histochemical methods. The levels determined in the three groups were quantitated and compared by computer-imaging analysis. The results showed that in early stage of atherosclerotic process, the NOS decreased while the ET increased in aorta. In early stage of essential hypertension, however, both NOS and ET levels increased. These results suggest that the increased ET and the change of NOS may play a role in the pathogenesis of hypertension and atherosclerosis. PMID- 10684007 TI - [A comparison of chromosome fragile sites in elderly and young people]. AB - In this comparative study, the authors investigated the chromosome aberrations and expression of fragile sites in the lymphocytes which were collected from 45 elderly people and 29 young people and were cultured with FUDR for 24 hours. The results showed that the aberration rate was 24.75% for elderly people, but 4.22% for young people (P < 0.01): that the expression of fragile sites was 45.94% for elderly peoples, but 23.02% for young people (P < 0.01): and that the relation of aberrations with fragile sites was 79.98% for elderly people, and 92% for young people. These findings suggest that chromosome aberration is in close association with fragile site. PMID- 10684008 TI - [Study on the relationship between occupational factors and work ability of middle aged workers]. AB - Based on the assessment of work ability index (WAI) of middle aged workers, associations between occupational factors and work ability decline of middle aged workers were examined in a case-control study. 1037 workers (age ranged from 40 to 60) were included in the study. The study group comprised 180 workers of mean age 47.71 years representing those who's work ability decreased. The control group comprised 857 workers of mean age 47.22 years representing those who's work ability was on a normal level. Results showed that some common occupation hazards such as dusts, noise, vibration, humidity, and high temperature were more frequently occurred in work environment of the study group than that of the control group. High risks were found in workers with these hazards. Another significant risk factor of work ability decline was heavy physical load. Significantly elevated risks were observed for physical load perception(OR = 2.52), repetitive work (OR = 1.36), poor work postures(OR = 1.48), and carrying heavy loads(OR = 1.59). As compared to physical loads, mental loads were not in relation to work ability decline. The information about occupational factors affecting work ability will be helpful for redesiging the work for the middle aged workers. PMID- 10684009 TI - [Characteristics of F-V curve configuration and its tangent time constants and its wave-speed mechanism in normal humans above 50 years old]. AB - To understand the characteristics of F-V curve configuration and its tangent time constant (tau 1) in normal humans above 50 years old, the tau 1 at high, mid, low lung volumes (tau 175, tau 150, tau 125) were measured from F-V curves for 135 normal humans aged 50-84. The results showed that the convex type was the most frequent one (40%-75%) among the 4 types of F-V curve and its tau 1 characteristics were tau 175 > tau 150 < tau 125 and magnitude of tau 175 < 1s; on the other hand, the concave type was the least frequent one (0.18%). These suggested that the convex type and its tau 1 characteristics might be the changes in senility. Further, the mechanical mechanism for the convex type of F-V curve configuration was explored. PMID- 10684010 TI - [Relationship between tumor cell proliferating activity and biological behavior, prognosis in laryngeal carcinoma]. AB - Using PC 10, an antibody to proliferating cell nuclear antigen (PCNA) and a standard immunohistochemical staining the authors examined 11 cases of simple hyperplasia of epithelium (SHE), 32 cases of atypical hyperplasia of epithelium (AHE) and 42 cases of laryngeal squamous cell carcinoma (LSCC) for expression of PCNA, a protein associated with DNA polymerase dalta and DNA replication, a marker for tumor cell proliferation. The results revealed that PCNA indices in SHE, AHE and LSCC were 9.5%, 27.33% and 68.05%, respectively. The PCNA indices were 63.68%, 69.57%, 71.18% in the well, moderately and poorly differentiated LSCC, respectively, there were significant differences. The PCNA indices were 63.88%, 70.82%, 66.20% and 69.04%, respectively in stages I, II, III, and IV of LSCC; no significance was found. There was a strong negative correlation betwee survival time and the tumor cell proliferative activity (r = 0.6243). PCNA might provide a useful tool for studying cell proliferation in situ under normal and pathological condition. PMID- 10684011 TI - [Prognostic significance of P53 protein expression in patients with colorectal adenocarcinoma]. AB - In order to investigate the prognostic significance of P53 protein expression in the patients with colorectal adenocarcinoma, we used the P53 monoclonal antibody and ABC immunohistochemical method combined with the repairing method for antigen by microwave oven to study the relationship between the overexpression of P53 protein and the prognosis in 40 cases of colorectal adenocarcinoma. The results showed that 65% (26/40) cases of colorectal adenocarcinoma were positive for overexpression of P53 protein. The 5-year survival rate for positive cases was 50%, but 87.5% for negative cases. These findings suggest that the overexpression of P53 protein closely associates with the prognosis in colorectal adenocarcinoma (P < 0.05), thus it might become a new prognostic indicator. The prognosis in colorectal adenocarcinoma could be predicted thoroughly by considering the expression of P53 protein in combination with the traditional prognostic factors. PMID- 10684012 TI - [The role of lens epithelium in cataract formation in diabetic rats]. AB - This study was designed to evaluate the changes and the role of lens epithelium in sugar cataract formation, in regard to the fact that the highest level of aldose reductase is found in this layer of lens. By light and electron microscopy, we examined the histological changes of central epithelium in lens of rats made diabetic with streptozotocin (STZ) with or without AL1576, an aldose reductase inhibitor, at varying periods of time ranging from 5 to 40 days after intraperitoneal injection of STZ. Also, we examined Na-K-ATPase activity in lens epithelium of rats with diabetes, diabetes plus AL1576 and normal controls at the time of 30 days. The results showed that the first detectable abnormalities occurred after 15 days of STZ injection and were limited to the lens epithelium; cell edema, intracellular vacuoles and extention of rough endoplasmic reticulum pool were remarkable; that AL1576 could prevent almost all of the lesion mentioned above; and that Na-K-ATPase activity in lens epithelium of rats with diabetes increased at the time of 30 days. The findings suggest that lens epithelium may play an important role in sugar cataractogenesis. PMID- 10684013 TI - [Quantitive analysis of 5-hydroxytryptamine levels on edges of incised skin of rats]. AB - To find index for early diagnosis of antemortem wound, the authors used the ICS analysis in a quantitive study of 5-hydroxytryptamine(5-HT) levels on rat's incised-skin antemortem wound in comparison with the postmortem wound inficted at various intervals. The results showed that the 5-HT increased significantly at 5 min, reached a peak at 15 min and decreased at 60 min on antemortem wounds. The 5 HT levels of antemortem groups were prominently high (more than 40%, P < 0.05) as compared with those of postmortem group and the levels of 5-HT on the wounds of postmortem groups were higher (35.77%, P < 0.05) than those of controls as well. The authors suggest that in the cases where the levels of 5-HT are as high as 40% on the edges of the injured skin, the injury should be considered an antemortem wound. PMID- 10684014 TI - [Morphologic changes of red blood cells in various conditions under scanning electron microscope]. AB - To study the changes of RBC in various conditions, blood clots elapsed 6 days in water and air and bloodstains stored in variant time were observed under Scanning Electron Microscope (SEM). The results showed that the morphology of RBC elapsed 6 days in water and air seemed to retain their integrity. They were measured 5, 48-5, 88 microns in diameter, whereas the RBC in the bloodstains stored 14 years were obviously disfigured and some appeared membrane cracking. However, the structure of RBC membrane was well recognized under the SEM. The morphological changes under the SEM and the forensic significance of the RBC were discussed. PMID- 10684015 TI - [Applied anatomy of the vascularity in the ileocolic region and its clinical significance]. AB - To understand the characteristics of the patterns of arteries and veins distributed over the ileocolic segment, 50 cadavers were studied by gross anatomy. The arteries, veins and their paracolic anastomoses distributed over ileocecal region, ascending colon and transverse colon were observed. The results showed that the distribution pattern of venous vessel was far more constant than that of artery, that the arrangements of artery in the ileocolic segment were classified into 7 types, and there were no interruption of paracolic anastomoses between arteries. But in 3 specimens the paracolic anastemoses between the right colic vein and the middle colic vein were completely interrupted. These findings may be useful in guiding clinical practice and preventing postoperative complications in ileocolic replacement of esophagus. PMID- 10684016 TI - [The effect of partial hepatectomy and portal vein occlusion on aminopyrine metabolism capacity of the liver in rats]. AB - To observe the variation of metabolic function and anatomical weight of the liver after partial hepatectomy and occlusion of portal branches feeding the same magnitude of the liver lobes, hepatic 14C-aminopyrine demethylation capacity determined by aminopyrine breath test and liver weight in rats were examined and compared in series in this study. The results showed that 14CO2 expiratory clearance rate was reduced by 50% at 2 hours and by 70% at 24 hours after 70% hepatectomy. Then it increased gradually and was similar to the preoperative level in 2 weeks. The remnant liver weight was lowest at 2 hours and increased to 80% of the control in 7 days after 70% hepatectomy. It recovered to normal in 2 weeks. In 90% hepatectomy group, 14CO2 expiratory clearance rate decreased by 90% at 2 hours after operation. Thereafter, all rats died very soon. In the 2 portal vein occlusion groups, 14CO2 expiratory clearance rate had little change at 2 hours and it declined by about 40% at 24 hours after ligation of portal branches feeding 70% or 90% liver mass. It recovered gradually and returned to the preoperative level in 2 weeks. The total liver weight in 2 portal occlusion groups was not significantly different from that in the control except that it was slightly lower than the control at 24 hours after operation. This experimental study suggests that comparing deprivation of portal blood feeding partial liver mass with corresponding partial hepatectomy, the former can not only keep the anatomical liver weight relatively stable but also produce a lesser damage to hepatic metabolic function. PMID- 10684017 TI - [Respiratory muscle function and serum enzymology in hyperthyroidism before and after treatment]. AB - To investigate the effect of hyperthyroidism on respiratory muscle function and its possible mechanism, the thyroid function, serum enzymology, serum potassium, pulmonary function and respiratory muscle function were examined in 60 patients with Grave's disease before treatment and 26 patients among them after treatment, and 20 normal subjects as control. T3, T4, and FT4 increased while FVC and PImax, which reflect the respiratory muscle strength, and Pi/PImax, which reflects the reserve capacity of inspiratory muscle, decreased significantly in the 60 patients with Grave's disease, compared with the ones of normal subjects. The comparison of above measurements in the 26 patients between before- and after treatment showed that respiratory muscle strength increased obviously along with the improvement of throid function. The serum enzymology, potassium and TSH, however, were not abnormal and not changed after treatment. The thyroid functions in 10 patients with hyperthyroid heart disease were not different, compared with the ones of other 50 patients without hyperthyroid heart disease, but their respiratory muscle strength was significantly lower than the ones of latter. The above results suggested that hyperthyroidism could lead to significant decrease of respiratory muscle strength and its reserve capacity, whereas treatment for hyperthyroidism would improve respiratory muscle function, so the measurement of respiratory muscle function in hyperthyroidism cases might be useful in prediction of hyperthyroid heart disease. PMID- 10684018 TI - [Effects of fat emulsion on serum lipid, apo A I, apo B100, LCAT, KBR of postoperative patients with liver disorder]. AB - Twenty post-operative patients with liver disorder were divided randomly into two groups. Ten of them received mean 0.82 g.kg-1.d-1 fat emulsion(42% of the total calorie), and the other ten received mean 1.72 g.kg-1.d-1 fat emulsion (52% of the total calorie) via vein during 5 post-operative days. Their serum lipid, apoA I, apoB100, ketone body ratio (KBR), fat clearance, LCAT and so on were investigted. The results suggest that it is safe for liver-disordered patients with slight or even moderate abnormality of liver function to receive mean 0.82 g.kg-1.d-1 and 1.72 g.kg-1.d-1 fat emulsion after operation and the use of mean 1.72 g.kg-1.d-1 fat emulsion will do no harm but good to liver function. PMID- 10684019 TI - [A survey of prostate symptoms in old male population]. AB - To know well the prostate symptoms in old men, the prostate symptom scores of 412 male residents over 60 years of age in Chengdu area (116 in city and 296 in countryside) were investigated by using I-PSS. The results showed that there was a significant difference between the city and countryside in terms of symptoms score(P < 0.05) and quality of life scores (P < 0.05). No significant difference was noted among the three age groups (P > 0.05). However, the mean score and the percentage of high score section in the elder group were higher than those in the younger group. The prevalence of benign prostate hypertrophy (BPH) and the demands on quality of life in city were different from those in countryside, and the prevalence rate of BPH increased with age. PMID- 10684020 TI - [Effects of mifepristone on ICE expression and Fas expression in early pregnant chorionic villi]. AB - This study addressed the question whether mifepristone has any effect on apoptosis in early pregnant chorionic villi. Interleukin 1 beta converting enzyme(ICE) expression and Fas expression in early pregnant villi and their changes following mifepristone administration were detected by immunohistochmical method and computer image analysis. During early pregnancy, ICE and Fas were expressed in all component chorionic cells, predominantly syncytiotrophoblasts. ICE stained cytoplasma. Fas stained mainly cytoplasma, and in partial trophoblastic cells, Fas was expressed on membrane and nucleus. In the villi from the pregnant women who received mifepristone for 2 days, expression of Fas increased markely; the ratio of positive nuclear staining cells were significantly raised, but the immunostaining intensity of ICE was slightly increased. These results suggest that mifepristone may terminate early pregnancy via increasing the expression of Fas and promoting apoptosis in the chorionic villi. PMID- 10684021 TI - [Study on enzyme-linked immunosorbent assay for cholecystokinin]. AB - This study was undertaken to develop a method for assaying octapeptide cholecystokinin (CCK-8) using ELISA. The design of the method was based on the pretreatment of polystyrene microplates with ultraviolet irradiation and glutaraldehyde activation piror to coating. The results showed that this pretreatment let to stable attachment of CCK-8 to the solid-phase. Competitive ELISA with CCK-8 as a competitor gave excellent quantitive relationship. Using this method we distinguished two kinds of CCK-related peptides. This method proved to be simple, stable and reproducible, the intra- and inter-assay coefficients of variation were 4.75% and 7.80% respectively. PMID- 10684022 TI - [Application of antigen linked with chemicals on nitrate cellulose membrane for dot-enzyme immunoassay]. AB - HCMV-IgG in sera of pregnant women was detected by means of dot-enzyme immunoassay (DEIA), in which antigen was cross-linked on nitrate cellulose membrane through chemical reagent. The results showed better repetition by DEIA, compared with ELISA, and no significant differences were noted between the two methods in the rates of detecting HCMV-IgG in positive, weak positive and negative sera. The rate of agreement was 93.2%. In examining the sera which had been examined by ELISA for three times with the same, results the sensitivity, specificity, index and efficiency of diagnosis by DEIA were 100%, 96.6%, 196.6% and 98.7% respectively. Since this method is convenient, fast, of good repetition, economical and its diagnosis index meets the application criterion, it is more suitable for clinical laboratories. PMID- 10684023 TI - [DNA polymorphisms of apolipoprotein A I gene in Chinese endogenous hypertriglyceridemics]. AB - The restriction fragment length polymorphisms (RFLPs) of apolipoprotein(apo) A I gene were studied using polymerase chain reaction (PCR) in 69 endogenous hypertriglyceridemics (HTG) and 74 healthy subjects from a population of Chinese Han nationality in Chengdu area. The loci studied included Msp 1 within intron 3 of the apo A I gene; Xmn I, 5' to the apo A I gene; and Pst I, 3' to the apo A I gene. The results showed that both in HTG group and control group M1, X1 and P1 alleles were the major alleles and homozyous M1M1, X1X1 and P1P1 genotypes were the most frequent ones. The frequencies of rare M2 and X2 alleles in Chinese were significantly higher than those in European Caucasians (0.293 vs 0.111, P < 0.0006, 0.286 vs 0.130, P < 0.004), but no differences were found in the frequency of rare P2 allele (0.071 vs 0.046, P > 0.05). The frequency of rare M2 tended to increase in the HTG group when compared with the control group (0.353 vs 0.275, P > 0.05). Both in the HTG group and control group, subjects with genotype M2M2 had a higher serum mean concentration of TC, apo C II and higher serum TG/HDL-C ratio compared with the subjects with the genotypes M1M1 and M1M2 (P = 0.05, P < 0.03, P < 0.04), and the serum TG, apo C II and apo E levels had a tendency to increase. The subjects with the genotypes P1P2 and P2P2 had a higher serum apo A II levels compared with the subjects with the genotype P1P1 (P < 0.04). The data show that the only Msp I RFLP within the intron 3 of apo A I gene is associated with endogenous hypertriglyceridemia to some extent in Chinese population. PMID- 10684024 TI - [Immunoblots of hydrophobic OmpL39 of Leptospiral interrogans with immunoprotective Mb E4B7G5]. AB - The immunoprotective Mb E4B7G5 against outer membrane antigens from L. interrogans serovar Lai strain 017 were produced and used in immunoblots of the OMP of six strains of L. interrogans (017, 601, 603, 609, 620 and 245). The OMP from the six strains, which partitioned into the hydrophobic detergent phase, contained four-seven major proteins bands of 66 kd-16 kd. It was found that Mb E4B7G5 recognized only specifically the 39 kd antigenic band of strain 017, 601, 603 and 609, and did not recognize apparently any bands of strains 620 and 245. The findings suggest that Mb E4B7G5 be valuable for separating protective antigen of OMP and studying genetic vaccines. PMID- 10684025 TI - [Microvasculature of islets in human pancreas]. AB - The microvasculature of islets was studied by light microscopy with ink-injection and scanning electron microscopy with vascular casts respectively in 11 human pancreas. The results revealed that most of the islets were supplied by the branches of the intralobular artery directly. The islets appeared in 3 types (central type, peripheral type and mixed type) characterized by the site where the afferent vessels entered the islets. Two (convergent and continuous) patterns of portal vessels were found according to their calibre and length. Some of the small islets were supplied by the efferent vessels of the adjacent large islets, and the vessels connecting the large and small islets were called insulo-insular portal vessels. PMID- 10684026 TI - [Acupoints specificity in promoting the plasticity of cat spinal lamina II with acupuncture-quantitative EM study]. AB - Our previous studies have shown acupoints stimulation might promote the plasticity of spinal cord. In order to explore the acupoints specificity of the effects we used ten cats with unilateral dorsal partial rhizotomy (L1-S2, except L6). The rhizotomy sides of five cats received acupuncture stimulation (acupoints stimulation group) at Zusanli (St36), Xuanzhong (GB 39), Futu (St32) and Sanyinjiao (Sp 6) in the innervated areas of L6 spinal nerve for two courses, 10 days per coure. The rest cats were subjected to needling at the areas adjacent to the acupoints (nonacupoints stimulation group). Quantitative EM method was applied to detect (the population of synaptic terminals in lamina II 30 days postoperation. According to the number of synaptic contacts between synaptic terminal and postsynaptic element, synaptic terminals were divided into two types: the simple terminal (ST) coming from spinal and descending tract, and the complex terminal (CT) from dorsal root. The average numbers of the two types' synaptic terminals per photo were compared directly because the acupoints and nonacupoints groups' transverse areas of lamina II showed no significant difference. The results showed: (1) There was no significant difference for ST number between two groups. Associated with our previous study that acupoints stimulation did not influence ST number, the present result indicated that nonacupoints stimulation had no effect on ST number, (2) The complex terminal numbers were statistically different (P < 0.05). The percentage of CT in the nonacupoints group was only 53% of that in the acupoints group. This result was similar to two other nonacupuncture groups in our laboratory (the percentages were 59% and 62% respectively). It indicates that the effect of nonacupoints stimulation is similar to that of nonneeding, i.e. nonacupoints stimulation does not promote the plasticity of spinal lamina II or there might be acupoints specificity in promoting the plasticity of spinal laminal II. PMID- 10684027 TI - [L-arginine and nitric oxide have effects on glomerulus hyperperfusion of early diabetic rats]. AB - We used streptozotocin (STZ) to induce the animal model of diabetes mellitus in rats. On the 7th and 14th days of induction, kidney disorder, the levels of NO3- in plasma and tissue homogenate in different periods were observed. The NO3- levels in relation to the application of L-arginine (L-Arg) were also noted. The results showed that the kidney weight/body weight ratio significantly increased in different periods (P < 0.05). On the 7th day profuse proteinuria appeared and it markedly increased on the 14th day: creatinine clearance rate (Ccr) significantly raised, too (P < 0.005). The NO3- levels in plasma and tissue homogenate were getting higher with time. The level of NO3- significantly increased after L-Arg was perfused. It indicated that kidney disorder was present at the early stage of diabetes, and the raised Ccr indirectly indicated the increase of glomerular filtration rate. These suggest that NO3- and L-Arg may be important mediums which have effects of hyperfiltration and hyperperfusion of glomerulus. PMID- 10684028 TI - [Experimental study of hypoxic pulmonary hypertension induced by nitric oxide in rats]. AB - This experiment in rats was designed to investigate the effect and mechanism of nitric oxide (NO) in the induction of hypoxic pulmonary hypertension. The plasma concentrations of NO in normal controls and in the 1-, 2- and 3-week hyporemic ventilation groups were measured. The hemodynamic and pathological changes were observed in rats of the 2-week group after bolus injection of L-Arginine and NG nitro-L-arginine. The results showed that NO concentrations of the 1-2- and 3 week groups were 5 +/- 2.67 mumol/L 2.1 +/- 0.41 mumol/L and 0.5 +/- 0.16 mumol/L respectively, which were significantly lower than the control group's 6.73 +/- 1.83 mumol/L (P < 0.05). Bolus injection of L-Arginine 100 mg.kg-1.d-1 could relieve chronic hypoxic pulmonary hypertension and decrease the thickening of pulmonary arteries, but L-NNA could antagonize the effect of L-Arginine. This experiment demonstrates that chronic hypoxemia may decrease the release of NO and result in pulmonary hypertension. L-Arginine may be used to relieve pulmonary hypertension, but L-NNA may antagonize the effect of L-Arginine. PMID- 10684029 TI - [Effect of different hypoxic duration on changes of pulmonary circulation]. AB - In order to assess the relationship between the hypoxic duration and pulmonary hypertension, we investigated the pulmonary hemodynamics and right ventricular hypertrophy and pulmonary vascular structure changes in rats with hypoxic exposure for 1 day to 3 weeks. After hypoxic treatment, the pulmonary artery pressure of rats increased in 3 days, and it was up to peak at 2 weeks later. The weight of right ventricle of rats significantly increased after 1 week. The endothelium of pulmonary vascular of rats became swollen in 3 days, and then increased in extracellular matrix deposition. Vessel wall thickening was found in 1 week hypoxic rats, and it was very significant in 3 week hypoxic rats. The results suggest that the duration of hypoxic treatment is related to the change of pulmonary circulation in 3 weeks in rats. PMID- 10684030 TI - [Study on PHSAr gene expression of HBV in normal human hepatocytes]. AB - Based on the sequence of Hepatitis B Virus (type adr) (HBV) Pre S2 region, we synthesized a pair of PCR primers. After a test of specificity, the primers were employed to amplify the postulated transcripts of PHSAr gene in the liver samples of 3 normal human subjects. By RT-PCR and RNA-PCR, all amplification results were positive, while those from corresponding skeleton muscle controls were negative. It suggests that HBV Pre S2 region should be to some extent homologous to PHSAr gene exon(s). It also indicates that PHSAr gene is in expression at transcriptional level, but not in expression in tissues (e.g. skeleton muscle) un susceptible to HBV infection. Our data further confirm the role of PHSAr in HBV infection of hepatocytes, compared with those from membrane receptor researches. PMID- 10684031 TI - [The expression of tumor suppressor gene p53 and Rb gene in nasopharyngeal carcinoma]. AB - In order to observe the relation of tumor supperssor gene p53 and Rb gene to nasopharyngeal carcinoma, we investigated p53 gene mutations in exon 7-8 in 33 cases of nasopharygeal carcinoma (NPC), using single-strand conformation ploymorphism analysis of PCR (PCR-SSCP). The data showed p53 mutations in 7 of 33 specimens at exon 8 (21.2%). No mutation in exon 7 was detected. Analysis of Rb gene in NPC tissues by southern blot revealed the deletion and loss of activity of Rb gene in 11 of 13 cases (86.6%). Our results suggest that Rb gene and p53 gene are closely associated with the tumorigenesis of NPC. PMID- 10684032 TI - [Relationship of primary pulmonary carcinoma with P16 protein expression]. AB - This study sought to address the relationship of primary pulmonary carcinoma (PPC) with P16 protein expression. The expression of P16 protein in 65 cases of PPC was examined with immunohistochemical staining. The rate of loss of P16 protein expression in PPC was 36.92% +/- 18.45%. The rate of loss of P16 protein expression in adenocarcinoma (AC) was 28.47 +/- 16.33%, that in squamous cell carcinoma (SCC) was 35.95% +/- 17.36%, and that in adenosquamous cell carcinoma (ASCC) 57.88% +/- 10.18%. There significant differences were in the rate of loss of P16 protein expression between those aged < 60 with SCC and those aged > 60 with SCC, between low differentiated PPC and intermediate differentiated PPC, between AC and ASCC, and between SCC and ASCC (P < 0.05). There were no significant differences in the rate of loss of P16 protein between males and females with PCC, between left and right PPC, and between PPC and metastatic pulmonary carcinoma of lymph node (P > 0.05). These suggest that P16 protein expression may be one of the important standards in estimating the prognosis of patients with PCC. PMID- 10684033 TI - [Successful 48-72 h cold storage of dog kidneys with HX-1 solution]. AB - HX-1 solution was used in this study to determine its effects on long term kidney preservation. 20 female and male mongrel dogs were randomly divided into control (HC-WCU solution) and experimental (HX-1 solution) groups. Kidneys were flushed with HC-WCU or HX-1 solution and stored at 0.4 degree C for 48 or 72 h. After being preserved, the kidney was implanted in the right groin of the animal and anastomosed to the iliac vessles. This procedure was immediately followed by a contralateral nephrectomy. The experimental findings were: 1. The maximum serum creatinine levels were 642.60 +/- 158.60 mumol/L and 686.20 +/- 154.04 mumol/L respectively in HC-WCU solution subgroups for 48 or 72 h cold storage. But the maximum serum creatinine levels were 448.32 +/- 36.90 mumol/L and 524.60 +/- 109.38 mumol/L respectively in HX-1 solution subgroups. 2. 80% of the kidneys were viable after 48 h storage in HC-WCU kidney solution, but 100% viable when stored in HX-1 solution: 40% of the kidneys were viable after 72 h storage in HC WCU solution, but 60% viable when stored in HX-1 solution. 3. Histologically the kidney of dogs that died of renal failure was damaged worse in HC-WCU solution group than that in HX-1 solution group. These results suggest that HX-1 solution is superior to HC-WCU solution for preserving kidneys and HX-1 solution may store up the kidney of dogs to 48-72 h. PMID- 10684034 TI - [Expression of epidermal growth factor receptor in human duodenal ulcer]. AB - Expression of epidermal growth factor receptor (EGFR) was studied immunohistochemically in 111 cases of human duodenal bulb biopsy. The results showed that the expression of EGFR was strong on the luminal side of epithelial cell membrane, moderate in the basolateral membrane of epithilial cells and the cells of muscularis mucosae, low in the cytoplasm of the epithelial cells, and lower in the mesenchyma and Brunner's gland. No expression of EGFR was found in lymphoid aggregates. The expression was evidently stronger in duodenal ulcer and duodenal ulcer scar. EGFR expression was correlated with the severity of inflammation, but was not related to sex and age. In conclusion, the result suggests that EGFR is correlated with mucosal adaptation to inflammation, ulcer healing and tissue reconstruction. PMID- 10684035 TI - [Histopathologic and immunochemical changes in Sjogren's syndrome]. AB - To research into the causes of sjogren's syndrome, we investigated the lacrimal gland tissues of 6 cases of sjogren's syndriome by using electronmicroscopy and immunochemical methods. The results revealed that at the early stage the lacrimal gland cells showed degeneration, irregular arrangement and many intracellular vacuoles. At the middle stage, some lymphocytes infliltrated into the gland tissues. And at the advanced stage, lymphocytes and collagenous fibers were there in substitution of gland cells. PMID- 10684036 TI - [A study on interleukin-2 receptor genes in acute myeloid leukemia cells]. AB - To investigate IL-2 receptor (IL-2R) gene expression in acute myeloid leukemia (AML) cells and the role of IL-2 in AML, the IL-2R alpha mRNA and IL-2R beta mRNA in leukemic cells obtained from bone marrow of 41 cases with AML before chemotherapy were measured by reverse transcriptase-polymerase chain reaction (RT PCR). The responses to recombinant human IL-2 (rhIL-2) of these leukemic cells expressing IL-2R mRNA from 8 cases were assayed by 3H-thymidine incorporation. The IL-2R alpha mRNA and IL-2R beta mRNAs in leukemic cells were detected in 28 cases (68.3%) and 32 cases (78.0%), respectively. The IL-2R alpha mRNA and IL-2R beta mRNA in leukemic cells of 21 cases (51.2%) were expressed simultaneously. The responses to rhIL-2 of these leukemic cells from 8 cases were heterogeneous. The responses to rhIL-2 appeared in three types: proliferation, inhibition and no reaction. Only the cells of one case with M5 expressing IL-2R alpha mRNA and IL 2R beta mRNA responded proliferatively. The results demonstrate that the expressions of IL-2R genes in AML cells are a general phenomenon. The responses of these leukemic cells to rhIL-2 in vitro are heterogeneous. PMID- 10684037 TI - [Maturation of bone marrow megakaryocyte in patients with chronic idiopathic thrombocytopenic purpura]. AB - Colony formation units of megakaryocyte of twenty four patients with idopathic thromboeytopentic purpura (CITP) were observed by plasma clot cultures in vitro. We observed the maturation degree of megakarycyte and the effect of recombinant interferon alpha-2a (rIFN-alpha-2a) on megakaryocyte colony growth and maturation. The results showed that the number of CFU-MK of the patients with CITP was greater than that of the control group, while the patients with normal number of megakaryocytes on bone marrow smears had less BFU-MK and total clonies than the controls had. Through image analysis, we found that the black level of the positive cells of GP II a and GMP-140, the diameter and the area of megakaryocyte of patients with CITP were lower or smaller than those of the control group. These evidenced the block of megakaryocytopoiesis and megakarycyte maturation. The growth of MK colonies of marrow of patients with CITP was inhibited by r-IFN-alpha-2a. PMID- 10684038 TI - [The relationship between female infertility and genital inflammation of Chlamydia trachomatis]. AB - To address the relationship between female infertility and genital inflammation of chlamydia trachomatis (CT), we took samples from the cervix and fallopian tube of 147 cases of female infertility and investigated the rate of CT infection by using polymerase chain reaction (PCR) assay. 60 cases of normal pregnant women were chosen as controls. The results showed that in the infertile group the CT positive rates for fallopian tubes and cervices were 15.66% and 22.45% respectively, whereas in the control group the tubal positive rate was 3.33%. There was significant difference between the two groups in tubal positive rate (P < 0.05). Youngsters and workers were the high risk population of genital infection of CT. These results indicate that genital infection of CT is an important factor causing infertility; in that disease, the tubal infection of CT is even more important. Besides the damage to tubal mucosa and smooth muscle, CT infection causes tubal adhesion and obstruction. As a result it impairs the function of the fimbriae and leads to infertility. PMID- 10684039 TI - [Application of spectrophotometry to evaluation of the levels and the influence of chemotherapeutic drugs on ATP of malignant cell lines]. AB - To evaluate the value of ATP spectrophotometry for malignant tumor in vitro chemosensitivity test, Hela and L929 cell lines, which were derived from human cervical carcinoma and mouse fibroma respectively, were investigated. The results showed that when the cellular numbers were > or = 2 x 10(5), the linearity between the ATP concentrations and the cellular numbers was observed, with all 2 cell types studied, and that when L929 cells were incubated with cis-platinum (1x, 2x, 5x x plasma peak concentration, PPC), a marked fall in ATP concentrations was observed and a clear dose-response curve was obtained, but no decrease of ATP concentrations was measured after incubation with adriamycin (1x, 2x, 5x x PPC). This study suggests that ATP spectrophotometry is a cheap, feasible method for studying the energy metabolism and the influence of chemotherapeutic drugs on ATP concentrations of malignant tumor. PMID- 10684040 TI - [Investigation of potency to produce interleukin-2 of the peripheral blood T lymphocytes in patients with hemorrhagic stroke]. AB - For further understanding the changes in immune function of the patients with hemorrhagic stroke, we investigated the activity of interleukin-2 (IL-2) in the peripheral blood T lymphocytes from 32 patients. The results showed that the levels of IL-2 in the patients were lower than those of the healthy controls, but no significant difference was observed between the cerebral hemorrhage group and subarachnoid hemorrhage group. It suggests the cellular immune function of patients at the acute period is low. We consider that IL-2 might be of use as a kind of therapeutic measure to modulate patients' immune function and treat the diseases. PMID- 10684041 TI - [Development of an expert system for assessment of criminal responsibility capacity of mentally ill offenders]. AB - The objective of this study was to develop an expert system for the assessment of legal capacity (ESALC). Visual Basic for Windows 3.0 was employed in establishing ESALC, and 281 cases were assessed by the system. The results were in substantial agreement with those of expert assessment. Kappa = 0.68 (P < 0.01); accuracy 90.04%, sensitivity, 85.63% and specificity 96.49%. ESALC can be used as a supplementary instrument in forensic psychiatric assessment. PMID- 10684042 TI - [The residue and degradation of herbicide ammonium 2,4-dichloro-6-nitrophenate in the system of wheat and soil]. AB - To study the residue and degradation of ammonium 2, 4-dichloro-6-nitrophenate (a herbicide) in wheat and soil, a field test was made in Wuxi Jiangsu of East China and Chengdu Sichuan of West Chain during 1985-1992. The results showed that ammonium 2,4-dichloro-6-nitrophenate was unstable with a half-life of 12-18 days in soil. The residue in wheat was less than 0.05 mg/kg on the field doses recommended. These results may be used as a basis for the hygienic evaluation of ammonium, 2,4-dichloro-6-nitrophenate. PMID- 10684043 TI - [Primary study on the sensitivity of cytokinesis blocked micronucleus assay in CHL cells]. AB - Studies were performed to determine the cytochalasin B induced binucleated cell percentage influenced by clastogens and aneuploidgens and to compare the efficacy of cytokinesis blocked binucleated cells for scoring micronuclei with that of the conventional mononucleated method following the treatment with mitomycin C, methyl methanesulforate, colchicin and chloral hydrate. The results showed that mitomycin C decreased the binucleated cell percentage induced by cytochalasin B, whereas colchicin increased the frequencies of binucleated cells. The frequencies of micronuclei in binucleated cells were not significantly higher than those in the conventional mononucleated cells. The results suggest that cytokinesis blocked method is not more sensitive than the conventional method for scoring micronuclei. The factors that may influence the cytokinesis blocked micronucleus assay have been discussed. PMID- 10684044 TI - [The relationship between pulmonary function and work ability of aging workers]. AB - To explore the relationship between pulmonary function and work ability, a field study was conducted in 285 aging workers (aged 40 to 60) of different occupations in Chengdu area. The pulmonary function indices included FVC, FEV1, FEV1%, MMEF, and the pulmonary function index (PFI) which was the sum of the rating values from the first three indices. The work ability index (WAI) was also measured. Between work types, the indices, including FVC and FEV1%, showed significant difference (P < 0.05); FVC also showed significant difference between male and female workers (P < 0.05). There was a significant relationship between PFI (including FEV1% and MMEF) and WAI (P < 0.05), but the relationship between FVC (or FEV1) and WAI was not statistically significant (P > 0.05). Moreover, the agreement percentage between the classifications of PFI and WAI was 43.8%; the disagreement percentage was 4.56%. The results suggest that FEV1% and MMEF may be beneficial to the objective evaluation of the work ability of aging workers. PMID- 10684045 TI - [Electron spin resonance studies of the oxidative denaturation of hemoglobin of blood preserved at 35 degrees C]. AB - Studying the denaturation of oxidative hemoglobin of blood preserved at room temperature, it is helpful to overall understand erythrocyte injurious during blood preserved at room temperature. Using electron spin resonance technique, we have researched the denaturation of oxidative hemoglobin of blood preserved at 35 degrees C. We discovered that blood preserved at 35 degrees C, appears ESR absorption of high spin methemoglobin (g = 6) after preservation time is 36 hours. The high spin methemoglobin ESF absorption will increase when preservation time increases. When blood is preserved at 4 degrees C, it dose not emerge ESR absorption during 60 hours preservation. SOD activity will gradually decline in erythrocyte of blood preserved at 35 degrees C when preservation time increases. Oxidative denaturation of hemoglobin is probably caused with gradual declining of antioxidation system effect in erythrocyte. PMID- 10684046 TI - [Resistant antibiotic analysis of methicillin-resistant staphylococcus]. AB - Methicillin-resistant staphylococcus aureus is one of the important patholonic bacteria which cause nosocomial infection. In order to investigate the resistant antibiotic circumstances of this organism and hence provide foundations of prevention and treatment, we determined the resistant rates of 88 staphylococcus strains for methicillin and for other 14 kinds of commonly used antibiotics, and we determined the engendering beta-lactamase. The results showed that the isolation rate of methicillin-resistant staphylococcus aureus (MRSA) was 56%; the rate of methicillin-resistant staphylococcus coagulase-negative (MRSCoN) was 47.6%; the resistance rate and the multi-resistant rate of methicillin-resistant staphylococcus (MRS) for the 14 kinds of antibiotics were higher as compared with those of methicillin-sensitive (MSSA). Among all the tested strains, MRSA was the highest to engender beta-lactamase (92.9%); Methicillin-sensitive staphylococcus coagulase-negative (MSSCoN) was the lowest (39.4%). MRS was sensitive to vancomycin and furantoin. These suggest that when treating this bacterium, we should select these two antibiotics first. PMID- 10684047 TI - [Non-parametric testing for independence in data]. AB - In this paper, we have discussed the important effects of data independence on statistic results, and have explained in detainl the non-parametric testing for independence, which can facilitate medical researchers to use statistical methods for solving their problems. PMID- 10684048 TI - [Studies on the essential oil constituents of the stem bark of Magnolia obovata]. AB - For the extensive studies of the chemical constituents of Magnollis obovata, the essential oil from Magnolia obovata was obtained by steam distillation. The chemical components of the oil were examined by means of capillary gas chromatography and gas chromatography-mass spectrometry. 29 of the 75 separated constituents were identified. The content of each component identified was determined by area normalization method, which showed the content of 9 compounds was higher than 1%. PMID- 10684049 TI - [Optimization of quenching system for determination of emodin and chrysophanol by chemiluminescence method]. AB - Determination of the valid components in traditional Chinese medicine is one of the key steps for its identification, and the development of accurate and simple methods is necessary for the present analysis work of medicines. In this paper, the experimental conditions were optimized and procedures chosen for determination of emodin and chrysophanol by lunminol-H2O2-Cr3+ chemiluminescence quenching system. The dynamic range for determination of emodin was 10(-4)-10(-8) g/ml (r = 0.9965), with a detection limit of 1.57 x 10(-9) g/ml. The RSD and recovery for determination of 10(-7) g/ml of emodin (n = 6) were 0.4% and 92.8% +/- 0.5%, respectively. Satisfactory results were also obtained when the method was used to determine chrysophanol. This method was used, after separation by thin layer chromatography, to determine emodin and chrysophanol in several traditional Chinese patent medicines, and the results were in good agreement with those obtained by the standard colorimetric method. PMID- 10684050 TI - [Adsorption of permanganate onto activated carbon particles]. AB - In this paper, the adsorption behavior of permanganate onto activated carbon particles was studied as a function of pH, flow rate and the concentration of permanganate. It was found that permanganate was not adsorbed but reduced by activated carbon particles. The percent of reduction of permanganate was strongly pH-dependent. The reducing product of permanganate was mainly Mn2+ which was adsorbed onto the activated carbon particles by surface complexation. PMID- 10684051 TI - [Oxidative modification of serum LDL, VLDL and HDL induced by fed on high cholesterol diet in vivo in rabbits]. AB - Many lines of evidence suggest that LDL is oxidized in vivo and that ox-LDL is present in the artery wall. But the oxidation of VLDL and HDL in vivo has not been reported yet. A study on effects of high cholesterol diet on oxidative modification of LDL, VLDL and HDL in rabbits was made. The control group (n = 8) was fed on routine diet, and the experimental group (n = 8) on high cholesterol diet (routine diet supplemented with 5% lard, and 0.5 g cholesterol a day for each rabbit) for 12 weeks. The serum LDL, VLDL and HDL were isolated by the density gradient ultra centrifugation. The oxidative modification of LDL, VLDL and HDL was identified by agarose gel electrophoresis, absorbance at 234 nm and fluorescence of TBARS. The results showed that serum TC, TG and TBARS in the experimental group were significantly higher than these of the control group (P < 0.01). The electrophoretic mobility of LDL, VLDL and HDL was increased, and absorbance at 234 nm and TBARS of LDL, VLDL and HDL in the experimental group were significantly higher than these of the control group (P < 0.01). These results suggest that not only LDL but also VLDL and HDL were oxidatively modified in vivo in rabbits after fed on high cholesterol diet. PMID- 10684052 TI - [Serum apolipoprotein C II, C III and E levels in 437 male healthy subjects aged 40-70 in Chengdu area]. AB - Apolipoprotein (apo) C II, C III and E levels of 437 male healthy subjects aged 40-70 [with fasting serum triglyceride (TG levels < 2.26 mmol/L, total cholesterol (TC) levels < 6.21 mmol/L and plasma glucose levels < 6.10 mmol/L] in Changdu area were determined by radial immunodiffusion assay (RID). The results showed that the concentrations of apoC II, C III and E were 41.6 +/- 13.1, 112.8 +/- 31.0 and 38.7 +/- 8.2 mg/L (mean +/- s) respectively. No difference in the serum levels of apoC II, C III and E was noted between the men of different ages. The results of the linear correlation showed that there was a positive correlation of apoC II, C III, E with TG, TC, LDL-C; of apoC II, C III with BMI; and of apoE with ages. PMID- 10684053 TI - [The serum lipid and apolipoprotein levels of middle-aged male hyperlipidemics in Chengdu district]. AB - It has been evidenced that cardiovascular diseases (CVD) relate with many risk factors and serum lipids play and essential role in the development of CVD. In order to further study hyperlipidemia (HL) in the middle-aged males, we analysed the body mass index (BMI), fast blood sugar (FBS), serum lipids and apolipoproteins A I, A II, B100, C II, C III and E in 223 male HL patients aged 41-60 and 349 normal male subjects who matched the HL patients in age. The result showed that the increase of serum triglycerids (TG), was as might be expected, the major characteristic of the middle-aged male HL patients in Chengdu District. Besides the serum lipid and apolipoprotein B100, C II, C III and E levels, the BMI and FBS levels in the IIb, IV and V types of HL patients were significantly higher than those in the normal subjects. It also showed that in TG increased patients the increased percentages of serum lipids were significantly higher than those of apolipoproteins. The age distribution in HL patients revealed that only 25% of the HL patients were between 41 to 50 years old, while 50% in the 56-60 year-old group. The relationship between apolipoprotein levels and serum lipid metabolism is also discussed. The results suggest that the 51-60 year-old males should pay attention to diet and increase physical activities to reduce incidence of HL which is directly associated with CVD. PMID- 10684054 TI - [Relation of serum apolipoprotein B100 levels to lipids and apolipoproteins A I, A II, C II, C III and E levels in the middle and old age normal males in Chengdu area]. AB - This study was conducted in 438 normal men aged 40-70. The age groups were: 40 44, 45-49, 50-54, 55-59, 60-64, and 65-70. The results showed that the serum apolipoproteins levels (mean+/- s, mg/L) were: B100 levels 742.5 +/- 165.8, A I 1301.2 +/- 219.9, A II 299.4 +/- 49.0, C II 42.7 +/- 21.4, C III 113.3 +/- 34.0 and E 38.7 +/- 8.2. The levels of serum apoB100 increased with age. The serum apoB100 levels in the 5th and 6th age groups were significantly higher as compared with the groups 1, 2, 3 and 4 (P < 0.05 and P < 0.05). The fasting serum mean TG (1.39 +/- 0.45 mmol/L), TC(4.79 +/- 0.82 mmol/L) and LDC-C(1.62 +/- 0.40 mmol/L levels in the groups 5 and 6 were significantly higher as compared with the groups 1, 2, 3 and 4 (P < 0.01). Serum apoC II, C III and E levels were significantly increased in the groups 5, 6 (P < 0.05). The correlation analysis indicated that there was a positive correlation of apoB100 with serum TG, TC, LDL C, apoC II, C III and E respectively (P < 0.01); and a negative correlation with HDL-C levels (r = 0.1312); and apoB100 correlated negatively with apoA I (r = 0.0706). The results suggest that serum TG, TC, LDL-C, apoC II, C III and E are the main factors related with the serum apoB100 levels. PMID- 10684055 TI - [Construction of genomic library of L. interrogans serovar lai using lambda gt11 as the vector and a study of recombiant plasmid pDL121]. AB - A genomic library of L. interrogans serovar lai strain 017 has been constructed using lambda gt11 as the vector. DNA was partially digested by two blunt-end restriction enzymes, then methylated with EcoR I methylase; after EcoR I linker was added to the DNA, the linker-ended DNA was ligated to the dephosphorylated EcoR I digested lambda gt11 arms. The recombined DNA was packaged in vitro, and used to transduct E. coli Y1090 for amplification. There were 2.1 x 10(6) recombinant bacteriophages as recognized by their ability to form white plaques plated on Lac host in the presence of both IPTG and X-Ga1. A positive clone, designated lambda DL12, was screened with a rabbit anti-serum against L. interrogans serovar lai from the genomic library. The DNA from lambda DL12 was subcloned into plasmid pUC18. A recombinant (designated as pDL121) was obtained. SDS-PAGE analysis indicated that a 23 kd was expressed in E. coli JM 103 harboring pDL121. Western blotting analysis showed that a specific protein band molecular weight of 23 kd could be recognized by the rabbit antiserum against L. interrogans serovar lai strain 017. PMID- 10684056 TI - [Circadian rhythm of PGE2 level in plasma and condylar cartilage of young growing rats]. AB - This paper reports the circadian rhythm of PEG2 level in plasma and condylar cartilage of young frowing rats. The samples were measured at 0:00, 4:00, 8:00, 12:00, 16:00, and 20:00 respectively. The count of PGE2 were determined by RIA and analysed with simple Cosinor and coordination. The results revealed: (1) The levels of PGE2 in plasma and condylar cartilage of young growing rats fluctuated in strong circadian rhythm; (2) The rhythm of PGE2 in plasma between 16:00 and 24:00 was significantly different from that between 4:00 and 12:00, so for analysing PGE2 level in plasma, one should proper sampling time or adjust it to circadian rhythm; (3) The circadian rhythm of PGE2 in condylar cartilage could provide important reference for reatments in dentistry and orthopaedics. PMID- 10684057 TI - [Effects of estrone on transmission at neuromuscular junction of the toad]. AB - The effects of estrone on neuromuscular transmission were studied intracellularly at the muscle fiber endplate in isolated sciatic nerve-sartorius muscle preparations of the toad (Bufo bufo gargarizans) in vitro. The results showed that estrone (10(-4) mol/L), added to bath medium, caused an increase in amplitudes of endplate potentials (EPP) both in 10 micrograms tubocurarine treated and 10 mmol/L MgCl2 treated preparations, but the increase was more prominent in the former, and that estrone, 30 min after being added to bath medium, caused clear increase both in frequency and amplitude of miniature endplate potentials (MEPP), but the wave form of MEPP remained unchanged and the resting membrane potentials were also unchanged. These results suggest that estrone can increase neuromuscular transmission, which seems to involve primarily the presynaptic mechanism. PMID- 10684058 TI - [Effects of L-glutamic acid, gamma-aminobutyric acid and their respective antagonists on spontaneous discharge of nucleus paragigantocellularis lateralis neurons in rats]. AB - Using multibarrel microelectrode techniques, we studied the effects of iontophoretic application of L-glutamic acid (L-Glu), gamma-aminobutyric acid (GABA) and their respective antagonists DL-2-amino-5-phosphonovaleric acid (AP5), bicuculline (BIC) on the spontaneous discharge of the caudal half of nucleus paragigantocellularis lateralis (cPGCL) neurons (including respiratory related neurons) and the influences of AP5 and BIC on the effects of L-Glu and GABA respectively in 22 anesthetized spontaneously breathing Sprague-Dawley rats. The spontaneous discharges of all the cPGCL neurons tested were inhibited by GABA(n = 53). Most of the tested neurons were excitated by L-Glu (30/36). AP5 and BIC both showed three kinds of effects on the cPGCL neuronal spontaneous discharge; excitatory, inhibitory and no-effect. The excitatory effects of L-Glu and BIC and the inhibitory effects of GABA showed a dose-response relationship. AP5 could block partially the excitatory effect of L-Glu on a large part of neurons tested (14/19). BIC blocked, partially or completely, the inhibitory effect of GABA also on a large part of the neurons tested(18/24). The results implicate that there might exist endogenous L-Glu and GABA acting as neurotransmitters in the cPGCL area and excitatory amino acids (EAA, including NMDA and non-NMDA) and GABA receptors on cPGCL neurons. These neurotransmitters and receptors may mediate the regulatory action of cPGCL on respiration and other functional systems. PMID- 10684059 TI - [Alkaloid production of cultured coptis cells by two-stage suspension-culture]. AB - In this study the asexual cell line H292 induced and selected from Coptis gulinensis had rapid growth rate and could stably produce alkaloids. By the one stage method, after the cell suspensions were cultured on the same medium for six weeks, the increased dry and fresh weights of the cells were 20.96 g/L and 174.92 g/L respectively. The content of the total alkaloids in the cells was 14.79 mg/g cell dw. Per litter liquid medium could provide 323 mg alkaloid. In contrast, the cells were cultured by two-stage method. After having been cultured on the medium which contributed to the growth of the cells for three weeks, the cells were transferred to the medium which contributed to the production of the alkaloid and cultured for three weeks. Six weeks later, the dry and fresh weights of the cells were 16.72 g/L and 127.44 g/L, respectively. The biomass was lower than that by one-stage method, but the content of the total alkaloids was as high as 31.76 mg/g cell dw, which was much more than that by one-stage method. In addition, the content of the alkaloid in the medium was 25.31 mg/L. Per litter liquid medium could provide 556 mg alkaloid. The total yield of alkaloid obtained by two-stage method was 1.72 times than that by one-stage method. PMID- 10684060 TI - [Expression of metastasis-suppressor gene nm23H1 product in nasopharyngeal carcinoma]. AB - Using the specific monoclonal antibody of nm23H1 gene product and the immunohistochemical technique, we studied the expression of DNPK/nm23H1 and its correlation with lymph-node metastasis in nasopharyngeal carchnoma (NPC). The results revealed that, in a total of 31 cases of NPC, 13 showed positive expression with a positive rate of 41.9%. Among 21 cases of NPC with no lymph node metastasis, 11 (52.3%) showed positive expression while only 20% (2/10) of those with lymph-node metastasis were positive, and there was significant difference between them (P < 0.01). Meantime the expression was very low or not detectable in the cancerous cells of the match lymph-node metastasis and all 10 cases showed negative expression. Statistical significant difference was found between the group of NPC with no metastasis and the group of lymph-node metastasis (P < 0.01). The above results indicated that high expression of DNPK/nm23H1 existed in the group of NPC with no metastasis while low expression existed in those with metastasis and lymph-node metastasis. It suggests that the expression of nm23H1 is inversely associated with lymph-node metastasis. The DNPK/nm23H1 may play a role in polymerization of microtubulin protein and participate in the process of cancer metastasis. PMID- 10684061 TI - [PCR-SSCP analysis of p53 gene in human primary brain tumor]. AB - In this study PCR-SSCP alalysis was made to detect the mutation of the exon 5-7 of p53 gene in 37 cases of human primary brain tumor. The results showed that 10 human primary brain tumors had mutation, of which 9 were gliomas (37.5%) and 1 was meningioma (7.7%), and that the higher the grade of tumor was, the higher would be the frequency of mutation. These results suggest that human primary brain tumor is related to p53 gene mutation and such mutation may be the major reason for human primary brain tumor genesis and malignancy. PMID- 10684062 TI - [Detection of p53 gene mutations in hepatocellular carcinoma]. AB - This study screened 32 cases of hepatocellular carcinoma (HCC) from Chengdu detecting HBV DNA. HBsAg and p53 mutations by using Southern blot hybridization, immunohistochemistry and polymerase chain reaction/restriction enzyme digest methods, respectively. The results revealed all the cases had been infected by HBV; the frequency of HBV DNA integration into HCC cell was 72%, and the positive staining for HBsAg 96.3%(26/27). Of the 32 cases, 8 showed nuclear staining of p53 protein (25%), no mutation of p53 gene at all in 27 nontumorours liver tissues was identified. The resluts suggest the inactivation of p53 function may play a significant role in the genesis of HBV-associated HCC; however, the largely negative p53 mutation results in the current study conversely indicate that hepatocarcinogenesis may even involve other comprehensive mechanisms. Further studies are worth doing to evaluate the possible contribution of HBV to the p53 mutation in HCC. PMID- 10684063 TI - [An analysis of acute leukemia cells TfR expression in children]. AB - There are a lot of transferrin receptors on the acute leukemia cells. We used the method of "Receptor Radioligand Assay" to determine the numbers of TfR binding sites on leukemia cells from 20 child patients with acute lymphocyte leukemia (ALL) and 9 with acute myeloid leukemia (AML). The results showed the numbers of TfR binding sites of AML cells were higher than those of ALL cells, and the numbers of complete remission group were lower than those of dead or relapse group. This indicated there might be a relationship between the TfR expression and the prognosis. PMID- 10684064 TI - [The anti-inflammatory effects of superoxide dismutase]. AB - This paper reports the anti-inflammatory effects of superoxide dismutase (SOD) from pig blood on inflammatory animal models. The experimental results have shown that SOD has significant anti-inflammatory effects. It inhibited carrageenin induced foot-edema and croton oil induced granulation tissue edema of rats. It also inhibited arthritis induced by egg serum and Freund's adjuvant in rats. The effects were significantly dose-dependent. PMID- 10684065 TI - [The effect of Tianma injection on cultured vascular smooth muscle cells proliferation]. AB - The proliferation of vascular smooth muscle cells (VSMC) has been thought to play an important role in the development of hypertension and atherosclerosis. The aim of this study was to investigate the effect of Tianma Injection and its major component Gastrodin on the proliferation of cultured VSMC. VSMC were cultured and randomly divided into the control, Tianma and Gastrodin groups. The results showed that Tianma Injection significantly decreased cell numbers dose dependently and time-dependently, and it inhibited 3H-TdR incorporation in VSMC. It also increased the content of PGI2 production in VSMC. However, no significant differences of the above measurements were observed between the Gastrodin and control groups. The suggest that Tianma Injection can inhibit VSMC proliferation and the effect possibly results from its increasing PGI2 production content, and that the inhibiting effect of Tianma is irrelated to Gastrodin. PMID- 10684066 TI - [A confirmatory analysis on the causation of osteoporosis]. AB - Bone width (BW, cm), bone mineral density (BMD, g/cm) and bone mineral density (BMD, g/square cm) are three indexes to the mineral content of the bone. Since the ways, methods and instruments measurement are usually not the same, exact measurement of the three indexes could not be assured. Thus the results of data analysis might not keep with the facts. We treated the three indexes as latent variables, set up a linear structural relation model (LISREL) and hence confirmed the causation of osteoporosis. PMID- 10684067 TI - [Application of serum bile acid chromatography to the diagnoses of liver diseases]. AB - In order to explore the specificity of serum bile acid (SBA) chromatograph in the diagnoses of different kinds of hepatosis, we investigated by means of gas chromatography the changes of serum bile acids in workers who exposed to hexogen or chloroethylene and in patients who suffered from hepatosis such as acute jaundice hepatitis, chronic active hepatitis, cirrhosis and liver cancer. The results revealed different disturbances of SBA occurring in the liver injuries induced by the two kinds of hepatotoxicant. Serum lithocholic acid (LCA), deoxycholic acid (DCA) and chenodeoxycholic acid (CDCA) in workers exposed to hexogen and wre significantly different from those of the control group respectively (P < 0.01, P < 0.01, P < 0.05); on the other hand, only serum LCA and DCA went up in workers exposed to chloroethylene (P < 0.0005, P < 0.001). The main changes both concentrated on the secondary bile acids. In acute jaundice hepatitis, chronic active hepatitis, cirrhosis and liver cancer, serum LCA, DCA, CDCA and cholic acids (CA) all went up in different degrees compared with the control group respectively (P < 0.005, P < 0.025, P < 0.005, P < 0.005). But no difference was noted among the 4 kinds of bile acids (P > 0.5), except that between CA and CDCA. These provided the evidence of the diagnosis and identification of clinical hepato-biliary diseases and occupational liver injures. PMID- 10684068 TI - [Effects of chronic hypoxia on calmodulin activity in rats]. AB - To investigate the effects of chronic hypoxia of calmodulin activity in rats, we divided 40 wistar rats into four major groups. One group acted as control. The other three groups were made hypoxic by placing them in an isobaric hypoxic chamber (O2 = 10%) for 1, 2 and 3 weeks respectively. Calmodulin activity was measured by Phosphodiesterase method. The results showed that the mean pulmonary arterial pressure and the pulmonary vascular resistance in rats were remarkably increased by chronic hypoxia. The total calcium levels and the calmodulin content in the lung tissue of rats were significantly elevated at 2-3 weeks of hypoxia exposure. However, the calmodulin content in the lung tissue of the rats exposed to hypoxia for 1 week also increased, but it was lower than those at 2-3 weeks of hypoxia exposure. Therefore, we conclude that Ca(2+)-calmodulin system may play a central role in mediating hypoxai-induced pulmonary hypertension. PMID- 10684069 TI - [Effect of methylphenidatum on inspiratory muscles function in patients with chronic obstructive pulmonary disease and its mechanism]. AB - To have a better understanding of the effect of methylphenidatum on inspiratory muscles function, we studied the respiratory force parameters of 70 patients with chronic obstructive pulmonary disease by intravenous infusion methylphenidatum in a randomized controlled clinical trial. The indices of respiratory force parameter included maximal inspiratory mouth pressure (MIP), maximal midinspiratory flow (MMIF), forced inspiratory capacity (FIC), maximal works of inspiration (Wimax) and airway occlusion pressure (P0.1), etc. Aminophylline and Nikethamidi were chosen as controls. The results showed that MIP, MMIF, FIC, Wimax, P0.1 and minute ventilation (Vr) were significantly increased after administration of methylphenidatum and aminophylline. There were no significant differences in MIP, MMIF, FIC and Wimax after administration of Nikethamidi, but P0.1 was significantly increased and the increase was higher than that after administration of methylphenidatum and aminophylline groups. We conclude that methylphenidatum can significantly improve the function of inspiratory muscles as aminophylline can do. PMID- 10684070 TI - [Changes of serum TNF-alpha level, t-PA activivty and PAI activity in patients with silent myocardial ischemia or silent cerebral ischemia]. AB - Tumor necrosis factor-alpha (TNF-alpha) level, tissue-typed plasminogen activator(t-PA) activity and PA inhibitor (PAI) activity were determined in three groups: (1) 25 NIDDM patients with silent myocardial ischemia (SMI) or silent cerebral ischemia (SCI); (2) 18NIDDM patients without SMI or SCI; (3) 20 age matched normal controls. Diagnosis of SMI or SCI was based on the finding of ischemic evidence by SPECT of myocardiotomograph or cerebrotomograph. All patients ECG and blood pressure were normal, and they had no history of clinical symptoms and signs of MI or CI. The result showed that the TNF-alpha level and PAI activity in the ischemia group were the highest and the t-PA activity in the ischemia group was the lowest, as compared with those in the other two groups respectively. It suggests that in NIDDM patients who have high TNF-alpha, high PAI activity, low t-PA, and even no symptoms and signs of MI or CI, anticoagulant therapy might be useful to prevent the progression of diabetic macroangiopathies. PMID- 10684071 TI - [The study on insulin secretion function of beta-cell in new diagnosed type II diabetic patients]. AB - To determine type II diabetes in association with alteration in beta-cell function, we studied 30 patients with newly diagnosed diabetes and 30 matched controls. The beta-cell 1st phase insulin response was measured by intravenous glucose tolerance test and the 2nd phase insulin response was measured by oral glucose tolerance test. The results revealed that in type II diabetes mellitus the pulse of 1st phase insulin secretion could not be found. The amount of 2nd phase insulin secretion of the patients was lower than that normal controls, and the patients' secretary pulse of insulin was not concomitant with glucose pulse. These findings again suggest it is the defect in beta-cell function, not the peripheral resistance an essential cause of overt type II diabetes. PMID- 10684072 TI - [Prevention of acute hepatic failure after "90%" hepatectomy by portal branch ligation in rats]. AB - In this study, ligation of portal branches feeding 90% liver mass was performed in rats. After the ligation, 10% liver mass with portal blood perfusion underwent a progressive hypertrophy. It gained weight against the control by about 5-fold at the 14th day. The measurement of incorporation rate of H3-thymidine showed that the DNA synthesis increased rapidly in the liver tissue keeping portal blood supply. It reached a peak at 24 hours and retumed to normal at 7 days after the ligation. The ligated liver lobes under-went an atrophy gradually. With the pretreatment of such portal branch ligation 2 weeks before, 80% rats survived from the resection of ligated liver lobes which composed of 90% preceding liver mass. In the control group of sham portal branch ligation, all rats died within a short period of time after 90% hepatectomy. This study demonstrated that a secondary extended hepatectomy following the ligation of portal branches feeding 90% liver mass could prevent acute hepatic failure induced by primary 90% hepatectomy in rats. PMID- 10684073 TI - [Uterine activity in active phase]. AB - This study was designed to understand the uterine activity (UA) in normal labor. 43 nulliparous women(33 in normal course of labor and 10 in abnormal course of labor) were in the active phase of spontaneous labor with cephalic presentation for vaginal delivery. An open-end fluid filled intrauterine catheter connected to a pressure transducer was used to measure the uterine contraction intensity, frequency and duration. UA was quantified by means of Montevideo Unit (MU) and Uterine activity integral(UAI). The receiver operator characteristic(ROC) curve defined the cut-off points of UA between normal labor and abnormal labor. The results showed that MU and UAI were correlated with the rate of cervial dilatation in the active phase of labor (r = 0.3734 and 0.3502 respectively, P < 0.05). There was a strong correlation between MU and UAI (r = 0.7173, P < 0.001). The mean of UA in normal labor as MU = 226(s = 68.45) mmHg/15 min. UAI = 901(s = 258.02) kPa/15 min. The minimum of UA to normal labor in MU was 170 mmHg/15 min, of UAI 650 kPa/15 min. The study suggests that quantification of UA is very useful in the management of labor, especially in the proper use of oxytocin in cases abnormal labor. PMID- 10684074 TI - [Evaluation on ISGP classification of endometrial hyperplasia in clinical application]. AB - To evaluate clinical application of International Society of Gynecologic Pathologists Classification of endometrial hyperplasia 424 patients with endome trial hyperplasia were treated in our hospital from Sept. 1989 to Feb. 1995. Our of these cases, 339 were simple hyperplasia, 28 were complex hyperplasia and 57 atypical hyperplasia. We made an analysis of the cases of complex and atypical hyperplasia in terms of the ISGP classification. The total rate of curettage was 63.5%. The agreement rate for pathologic diagnoses before and after operation was 78%. Under the age of 40, 18 patients had atypical hyperplasia and 15 of them underwent hysterectomy. Eight patients had complex hyperplasia and 3 of them underwent hysterectomy. Above the age of 40, 39 patients had atypical hyperplasia and 30 of them had uterus removed. Twenty patients had complex hyperplasia and 12 of them had uterus removed. These suggest that the ISGP pathologic diagnostic method is good to avoid overdiagnosis and overtreatment. Yet, diagnostic curettage is still the best means in diagnosis before operation. PMID- 10684075 TI - [Perinatal monitoring in intrahepatic cholestasis of pregnancy]. AB - The sensitivity of meconium stain in amniotic fluid for prediction of fetal well being in intrahepatic cholestasis of pregnancy(ICP) was evaluated. The study consisted of an ICP group(n = 30), and a control group (n = 30) and the umbilical arterial pH value(< 7.2) was used as a standard. The positive and negative predicttive valus of meconium-stained amniotic fluid in ICP group were 80.0%; the positive and negative predictive values in control group were 60.0% and 92.0% respectively. There was no significant difference (P > 0.05) between the two groups in positive and negative predictive values. However, the positive and negative predictive values of the two groups were high, which indicated that meconium-stained amniotic fluid related to fetal hypoxia. Moreover, the incidence of meconium-stained amniotic fluid in ICP was higher than that in control (40.0%: 16.70%, P < 0.05). Therefore, we suggest that the amniotic fluid of patients with ICP should be observed very closely. When meconium-stained amniotic fluid is discovered, delivery by cesarean section is imminent. PMID- 10684076 TI - [A three-month follow-up study of excimer laser photorefractive keratectomy for myopia]. AB - In order to evaluate the efficacy of photorefractive keratectomy (PRK) in the treatment of myopia, we summed up and analysed the 3-month follow-up data on 145 cases treated in our hospital. Prior to PRK, refractive dioptal was -1.5 to 11.5; visual acuity was less than 0.1 in 137 cases (94%) and less than 0.5 in 8 cases(6%). One month after operation, the mean refractive dioptal was +0.98 +/- 0.88; visual acuity increased to > or = 1.0 in 95 cases (66%) and to > or = 0.5 in 142 cases(98%). Three months after operation, the mean refractive dioptal was 0.13 +/- 1.09; visual acuity increased to > or = 1.0 in 104 cases(72%) and to > or = 0.5 in 135 cases(93%). There was no severe complication noted in all cases. The results demonstrate that PRK is and effective, safe and predictable treatment for myopia. PMID- 10684077 TI - [A study of 46 forensic autopsy cases of medical tangle]. AB - Forty-six forensic autopsy cases of medical tangle were investigated for understanding the characteristic of medical tangle cases and for comparing the consistency between clinical and pathological diagnoses. The age ranged from 6 days to 68 years. Women outnumbered men by ratio of 1.3:1. The most common occupation was farming. The consistency between clinical and pathological diagnoses was 34.78%. The causes of medical tangle and the difference between clinical and pathological diagnoses were discussed. PMID- 10684078 TI - [Quantitation of human apolipoprotein B100 by immunoturbidimetric assay]. AB - An immunoturbidimetric assay(ITA) for quantification of human serum apolipoprotein B100 (apoB100) was developed. The assay is sensitive, rapid, specific, easily automatic and thus convenient for routine work. The minimu measurable concentration of apoB100 was 3.1 micrograms in each assay. The standard curve with a working range of 0.34-1.79 g/L was plotted. The between and within assay coefficients of variation were 1.13%-3.48% and 2.91%-4.48% respectively. The recovery was 99.23% +/- 3.57%. The mean serum concentration of apoB100 in 100 normal subjects was 0.821 +/- 0.165 g/L. The correlation coefficient of RID for apoB100 to ITA was 0.8626(P < 0.001). PMID- 10684079 TI - [A newly developed equipment for cleaning of sample tube used in nuclear magnetic resonance experiment]. AB - Cleaning sample tubes used in nuclear magnetic resonance experiment is a time consuming work. We developed a new type of equipment based on fluidics for cleaning those sample tubes and successfully solved the problem. For several years of application, the developed equipment proved to be time saving and very effective. It is simple in design and easy to make up, and it can wash whatever glassware that is narrow in diameter and long in length. PMID- 10684080 TI - [Changes of the gene expression of vascular endothelial growth factor in the lung of rats with chronic hypoxic pulmonary hypertension]. AB - To observe the effect of chronic hypoxia on the gene expression of vascular endothelial growth factor (VEGF) in rat's lung, and the role of VEGF in the pathogenesis of hypoxic pulmonary hypertension, thirteen male Wistar rats were exposed to isobaric hypoxia for 3 weeks. The pulmonary artery pressure was measured by right cardiac catheterization. The serum level of VEGF was measured by Elisa. The VEGF cRNA was labeled with digoxigenin-UTP by in vitro transcription. The expression of VEGF mRNA in the lung was examined by hybridization in situ. The pulmonary artery pressure was significantly increased after hypoxic exposure. The serum level of VEGF in rats treated with hypoxia (420.3 +/- 73.1 pg/ml) was significantly increased in comparison with that of normal rats (322.2 +/- 58.1 pg/ml). The VEGF hybridization signals on the wall of pulmonary arteriole were significantly increased in rats with pulmonary hypertension. Chronic hypoxia can markedly increase expression of VEGF mRNA in the pulmonary arteriole and hence stimulate VEGF synthesis and secretion. The increase of VEGF may play a role in the developing process of hypoxic pulmonary hypertension. PMID- 10684081 TI - [Changes of level and distribution of vascular endothelial growth factor in the lungs of rats with hypoxic pulmonary hypertension]. AB - This study was designed to elucidate whether the level and distribution of vascular endothelial growth factor (VEGF) are changed in the lungs of rats with hypoxic pulmonary hypertension. 13 male Wistar rats were exposed to isobaric hypoxia for 3 weeks. The pulmonary artery pressure was measured by right cardiac catheterization. The level of VEGF in pulmonary homogenate was measured by Elisa method. The distribution of VEGF in the rat lung was examined by immunohistochemistry. The results showed that the pulmonary artery pressure was significantly increased after hypoxic exposure. The level of VEGF in pulmonary homogenate of rats treated with hypoxia (466.9 +/- 75.5 pg/g) were significantly increased as compared with taht of normal rats (376.2 +/- 47.1 pg/g). The contents of VEGF in the wall of pulmonary arteriole were significantly increased in rats with pulmonary hypertension. So we suggest that chronic hypoxia can strongly stimulate VEGF secretion, and VEGF may mediate the process of hypoxic pulmonary vascular remodeling and pulmonary hypertension. PMID- 10684082 TI - [Isolation and identification of smooth muscle cells from pulmonary artery in rats]. AB - To explore the method of isolating acutely the smooth muscle cells from pulmonary artery in rats, small pulmonary arteries (700-200 microns, ID) were dissected free of connective tissue and were allowed to digest in a N-2-hydroxyethyl piperazine-N'-2-ethanesulfonic acid(HEPES)-buffered physiological saline solution (HPSS) containing collagenase, papain and bovine serum albumin. The tissue was then triturated to disperse smooth muscle cells. The isolated cells in suspension were identified and photographed with film on electron microscope (EM). We succeeded in isolating the single smooth muscle cell, which appeared compressed typically. 90% cells in suspension were identified smooth muscle cells on EM. We conclude that the method for isolation of pulmonary arterial smooth muscle cells is simple, stable and effective and is recommanded for use. PMID- 10684083 TI - [Qualitative and quantitative analyses of telomerase activity in cultured cells]. AB - In this study, we established a qualitative PCR-TRAP assay by using SYBR Green stain instead of EB stain; this modification based on Kim's method raised the sensitivity of telomerase detection by 25-100 fold. Besides, a quantitative assay was established by us using 32P-ATP labeled TS primer and the internal control TSK1. We detected the telomerase activity of 12 cultured cells by means of these two assays. The results showed that the telomerase activity could be detected in all the cells, but the activity levels of the cells differed prominently (from 23 to 652 TPG units). The establishment of PCR-TRAP assay and especially the establishment of quantitative assay have enabled us to evaluate the telomerase activity of cells more accurately, and they can be used in our further studies of tumor gene therapy by reducing the telomerase activity of tumor cells. PMID- 10684084 TI - [Immunoprotection in guinea pigs using DNA recombinant plasmid rpDJt and expressed protein P68 in L. interrogans serovar lai]. AB - Immunoprotection against the infection by Leptospira interrogans serogroup Icterohemorrhagiae serovar lai strain 017 was demonstrated in guinea pigs vaccinated with DNA recombinant plasmid rpDJt and expressed protein P68 derived from genomic library of Leptospira strain 017. Thirty days after active immunization, each group received intraperitoneally (1/2 dose) and subcutaneously (1/2 dose) inoculum of L. interrogans serovar lai stain 017; cultures were adjusted to 5 x 10(8) cells/ml. All guinea pigs were observed for 10 days after challenge. Survival (%) of P68 group was 100(7/7); P23 group was 75(3/4); group rpDJt was 77(10/13); group lack recombinant (control) pT7-7 was 25 (3/12), and group with whole-cell inactivated vaccine was 93(13/14). Although the protective antigen in the leptospires has yet to be determined, it is evident that expressed protein P68 conferred a high degree of immunoprotection in guinea pigs. PMID- 10684085 TI - [A comparison study on the sensitivity of lung tumor short-term induction test in three strains of mice]. AB - To compare the sensitivities of lung tumor short-term induction in different strains of mice, the carcinogenicity risk was evaluated with the incidence rate and the average number of tumors. Three different strains of mice (KM, BALB/c and A/J) were injected with a single different dose of urethane i.p. The results demonstrated that the lowest dose for increasing tumor risk was 100 mg/kg in KM, 200 mg/kg in BALB/c, and lower than 25 mg/kg in A/J strain. So, the sensitivity of A/J strain is the highest and that of BALB/c is the lowest. PMID- 10684086 TI - [Effect of fluoride on proliferation and differentiation in rat and mouse embryo bud cell in vitro]. AB - The effect of fluoride on differentiation and proliferation of rat and mouse embryo limb bud cell were studied with micromass cultures in vitro. Embryo limb bud cells of rat (13-day) and mouse (12-day) were subjected to culture for 5 days. The results showed that fluoride could inhibit differentiation of cells without affecting cells proliferation. The concentrations of 50% inhibition of cell differentiation (ID50) were 6.8 micrograms/ml(rat) and 7.3 micrograms/ml(mouse). The concentrations of 50% inhibitions of cell proliferation (IP50) were 44.1 micrograms/ml (rat) and 63.6 micrograms/ml (mouse). The IP/ID50 values 6.4(rat) and 8.7 (mouse) were both greater than 5. According to the assessment criteria of Flint and Cheng Wanrong, the fluoride may be an embryo limb bud cells specific inhibitor. It could have potent teratogenicity. PMID- 10684087 TI - [Application of mouse limb bud culture to study the influence of zinc on teratogenesis induced by cadmium]. AB - In this study, an in vitro method of mouse limb bud culture in self-made rotator with continuous supplementation of gas mixture was employed in studying the teratogenic potential of cadmium and the influence of zinc on the teratogenesis induced by cadmium. Image analysis on the area and the form of the bone analgen of the cultured limb was used to evaluate quantitatively their teratogenic potentials. Different amounts of cadmium were directly added to culture medium. As cadmium concentrations were increased from 0.1 to 1.0 microgram/ml, the degree of morphogenetic differentiation and the area of the bone anlagen of limbs culture were significantly decreased. The paws and long bones were affected seriously. Cadmium had a greater effect on chondrogenic tissue than on soft tissue. Then various levels of Zn, together with cadmium (1.0 microgram/ml medium), were added into the culture media. As Zn concentrations increased from 1.0 to 10.0 micrograms/ml, the degree of morphogenetic differentiation and the area of cartilaginous bone anlagen of limbs culture were improved or increased. The long bones were better ameliorated as compared with the paw. PMID- 10684088 TI - [The effect of electroacupuncture stimulation an neurotrophic substance in cat spinal dorsal horn]. AB - For exploration of the mechanism of collateral sprouting from injured spinal cord, polyacrylamide gel electrophoresis (PAGE) and modified hanging drop culture method were used to examine the effect of acupuncture on the amount and the biological activity of neurotrophic substance in larger than 50 kd fraction of dorsal horn in cat subjected to partial dorsal rhizotomy. The results showed that the amount of protein in relative mobility (RM) 0.1 zone increased significantly in the acupuncture-operated group by PAGE and that of protein in RM 0.5 zone decreased significantly as compared with the unoperated group. While this data was compared with the data of another operated group in our lab, the amount of RM 0.1 protein of the acupuncture-operated group was significantly higher than that of the latter group; however, no obvious difference was noted between the two groups in the amount of RM 0.5 protein. It suggests that acupuncture may promote tha amount of RM 0.1 protein to increase, but acupuncture has no influence on the amount of RM 0.5 protein. Also, the results showed that, in culture, accompanying with the increase in amount, the activity of RM 0.1 protein for promoting DRG neurite-outgrowth increased. This indicates that the RM 0.1 protein has a function to promote DRG neurite-outgrowth, but the RM 0.5 protein has no obvious effect on DRG neurite-outgrowth. PMID- 10684089 TI - [Detecting expression of the multidrug resistance gene product (P170) in human tumor tissues and cells by flow cytometry]. AB - We have detected the human multidrug resistance gene (MDR1) product P170 in 29 solid tumor samples and K562 cell line through indirectimmunoflourence staining by flow cytometry using mouse monoclonal antibody (McAb). The results showed that the expression of P170 was detected in 18 samples, the positive ratio being 62.1%; the expression was not detected in 11 samples, the negative ratio being 37.9%; and 99.9% of K562 cells expressed P170. In 16 of the 18 positive samples, the percent ratio of tumor cells for expression of P170 was less than 30%; in the other 2, more than 30%. This indicated that the positive ratio of P170 of most tumor samples was high, but their percent ratio of P170 was low. Thus it provided a parameter for reference in evaluating the efficacy of clinical antitumor treatments. PMID- 10684090 TI - [Determination of valaciclovir polybutylcyanoacrylate nanoparticles]. AB - The valaciclovir content of polybutylcyanoacrylate nanoparticles was determined by HPLC after dissolving the valaciclovir-poly butylcyanoacrylate nanoparticles in a solvent mixture. ODS-C18 column was used. The mobile phase consisted of CH3OH-0.02 mol/L KH2PO4 (20:80). The linear range was 2.02-20.20 micrograms/ml; the recovery 97.30%, and RSD 4.90%. This method is accurate and can be used for determining the contents of other nanoparticles. PMID- 10684091 TI - [A study of immobilized enzyme-assisted semisynthesis of human insulin]. AB - In this paper, a study on preparation of human insulin by immobilized enzyme assisted semisynthesis is reported for the first time in our country. Porcine insulin with Thr(Bu(t)OB(t) catalyzed by immobilized trypsin has been converted into human insulin via a two-step transpeptidation. With the systematic technique of optimization, the rate of transpeptidation is 66%-70%. PAGE photometry is used to determine the conversion rate of semisynthesis. PAGE and C-terminal analysis of protein are adopted to identify the semisynthesis human insulin. PMID- 10684092 TI - [Improving the bonding strength of castable ceramic crowns]. AB - To study the influence of bonding agent on fracture strength of Plat-II castable ceramic crowns bonded to molars, we tested the specimens in vitro with Panavia 21 resin bonding agent and porcelite dual cure cement. The results showed that the fracture loading (845.8 +/- 137.1 N) of crowns bonded with Panavia 21 bonding agent was higher than that (534.0 +/- 58.7 N) of crows bonded with porcelite dual cure cement. The difference between the two agents was statistically significant (P < 0.05). The crowns bonded with Panavia 21 agent had higher strength than bite forces. These suggest that costable ceramic crowns could be bonded with Panavia 21 resin agent in clinical practice. PMID- 10684093 TI - [A biodistribution experiment on 125 I-VIP in mice]. AB - This experiment was designed to investigate the biodistribution characters of 125 I-vasoactive intestinal peptide (VIP) with high specific activity in normal mice and the effect of VIP on the distributive test. After intravenous injection of 125 I-VIP, the mice were killed during 180 min. Blood, lungs, liver, intestine and kidneys were collected respectively and measured in 7 counter, finally the measurements of their radioactivity (cpm) were converted into ID%/g tissue and the results were expressed as mean+/-s. The effect on VIP on distributive test was evaluated by simultaneously injection of 125 I-VIP (74kBq) which contained 10,20 and 40 microgram VIP respectively; the mice were killed at 5 and 20 min. respectively, the collection of tissues and the management of data were the same as above. Most of the 125 I-VIP was distributed in the lungs rapidly, the radioactivity was mainly eliminated through kindneys; the T1/2 of activity in blood was shorter than 20 min; the difference in activity between liver and blood was not significant after 20 min. (P >0.05); the activity intestine remained lower during the experiment. The uptake of 125 I-VIP in lungs, liver and intestine was inhibited by VIP in dose dependence. The rates of inhibitory effectiveness of 10, 20 and 40 microgram VIP in lungs were 45.12%, 56.64% and 68.12% respectively at 5 min., and 53.65%, 71.03% and 79.03% respectively at 20 min. The present study indicated that the uptake of 125 I-VIP in various tissues of the mice possesses the character mediated by VIP receptor, 125 I-VIP is mainly accumulated by lungs, eliminated through kidneys, and the hepatobiliary system cannot remove it. The biodistribution character of 125 I-VIP is helpful to 131I (123I)-VIP imaging in the detection of gastrointestinal tumor. PMID- 10684094 TI - [Effects of flunarizine and vitamin C on hemodynamics in rat heart subjected to ischemia-reperfusion]. AB - Langendorff perfusion isolated rat heart was subjected to total global ischemia (coronary flow rate is equal to zero) for 10 minutes and reperfusion for 15 minutes. The heart rate (HR), left ventricular developed pressure (LVDP), coronary flow rate (CFR), electrocardiogram (ECG) and the effects of calcium antagonist-flunarizine (FNZ) and/or oxygen free radical scavenger--vitamine C on the above parameters were observed. The results showed that FNZ dilated coronary vessel (P < 0.01) and had a slight negative chronotropic effect, but it had no effect on LVP. Vitamine C did not affect HR, LVP and CFR. The recovery of the product of HR and LVDP-Rate Pressure Product (RPP) in the FNZ + Vit. C group, Vit. C group and FNZ group was significantly higher than that in the control group (P < 0.05) at ten minutes reperfusion. All the results suggest that FNZ and Vit. C may improve the recovery of heart function after reperfusion. PMID- 10684095 TI - [Expression of glial cells line-derived neurotrophic factor in the central nervous system of rat]. AB - Glial cell line-derived neurotrophic factor (GDNF) iss a distant member of the transforming growth factor-beta, originally isolated by virtue of its ability to induce dopamine uptake and cell survival in cultures of embryonic ventral midbrain dopaminergic neurons, and more recently it has been shown to be a potent neurotrophic factor for motor neurons. However, the distribution of this molecule in the central nervous system (CNS) is not known yet. In the present study, the expression of GDNF of rat was investigated by using immunohistochemistry. GDNF immunoreactive fibers or bodies were found in molecular layer and Purkinje layer of cerebellum. In the spinal cord, neuronal expression was found widely, especially in ventral horn, Clarke's column. GDNF was also expressed in fiber terminals of I and II lamella of dorsal horn. Additionally, GDNA expression was found in the ependymal epithelium and neuroglia of spinal cord. The nuclei of neuroglia were stronger than perikaryon. The neurons in cerebral cortex showed very weak immunoreaction. We conclude that GDNF is expressed in many different cellular systems within the CNS, suggesting the multiple functions of GDNF in the adult CNS. PMID- 10684096 TI - [Regulative effects pentagastrin and somatostatin on growth of human gastric cancer cells in vitro]. AB - In order to observe the regulative effects of pentagastrin (PG) and somatostatin (SS) on the growth of two human gastric cancer cell lines (HGC803 and HGC823) in vitro, we observed the effects of PG and SS on proliferation of human gastric cancer cells by means of MTT. The contents such as gastrin, insulin, and glucagon were determined by radioimmunoassay (RIA), and the hexosamine content was determined by Neuhaus' method. The results showed that the growth of the two human gastric cancer cell lines were obviously promoted by PG. On the contrary, the growth and secretion of gastrin and glucagon were inhibited by SS. In addition, the hexosamine content of human gastric cancer cells was significantly increased by PG (7.58 +/- 0.66 versus 4.20 +/- 0.39 pg/cell, (P < 0.05). But the hexosamine content was decreased by SS (2.62 +/- 0.29 versus 4.20 +/- 0.39 pg/cell, P < 0.05). These findings indicate that the growth of gastric cancer cells is regulated by PG and SS, nevertheless a host of problems need to be elucidated. PMID- 10684097 TI - [An endometrium morphometry study on ovariectomized rats subjected to nilestriol and estradiol replalcement therapies]. AB - This study was designed to evaluate the effect of estrogen on rat endometrium and compare the results of Nilestriol and Estradiol replacement therapies for preventing osteoporosis in ovariectomized rats. Forty 4-month old SD female rats were randomly divided into four groups, namely sham operation, bilateral ovariectomy, ovariectomy plus supplementary ethinyl estradiol (0.2 microgram/100 g B. W. qd x 5), and ovariectomy plus supplementary Nilestriol (0.15 mg/100 g B. W. once a week). Computerized image analyzer was used to evaluate the endomitrial pathological changes 10 weeks later. The results showed an atrophied endometrium in ovariectomized group, a slightly proliferative endometrium in Nilestriol treated group, and a markedly proliferative endometrium with local atypical hyperplasia in estradiol treated group. The difference between group 3 and group 4 in morphometry was significant (P < 0.5). These suggest the relative safety of Nilestriol replacement therapy for preventing menopausal osteoporosis. PMID- 10684098 TI - [Clinicopathological and immunohistochemical study of hepatoblastoma]. AB - To investigate the relationship between immunoreaction of histologic subtype and prognosis, this paper analysed the clinicopathological data from 20 cases of hepatoblastoma. Immunohistochemical staining was performed in 18 cases. The results showed that cytopolasmic postivities of epithlial tumor cells were observed by CK, AFP, S-100 protein and vimentin in 14, 10, 9 and 4 cases respectively. Positive staining for CEA was seen in the nuclei of epithelial tumor cells in 11 cases. Nuclear P53 protein staining was found in 9 cases. Nuclear and cytoplasmic postivities of P16 protein were observed in 7 cases. S 100 protein, vimentin, CK and P16 protein were detected in mesenchymal component in 1 case. This study suggested that immunoreactions of hepatoblastomas were different in histologic subtypes. The expression may correlate with the neoplastic differentiation and prognosis. PMID- 10684099 TI - [Effect of ischemia/reperfusion in cardiac surgery with cardiopulmonary bypass on nitric oxide levels of peripheral blood in patients]. AB - To examine the effect of ischemia/reperfusion in cardiac surgery with cardiopulmonary bypass (CPB) on nitric oxide (NO) levels of peripheral blood, venous levels of NO2- and NO3- were measured at multiple time-points before, during, and after CPB by means of high-pressure liquid chromatograph to reflect NO concentration indirectly in 20 patients. The results showed there was a linear regression between the decreased trend of NO release at the period of peri-CPB is related to the endothelial cell activation caused by ischemia/reperfusion at the same time. Therefore it is necessary to explore the regulation and action of NO release during systemic inflammatory response procedure in CPB. PMID- 10684100 TI - [Quantitative analysis of motor unit potential in patients with dermatomyositic disease]. AB - To diagnose dermatomyositic disease well, we adopted the method of EMG in an investigation of 37 patients. Their electrophysiologic parameters of 220 muscles were determined. The results showed that the parameters, including the average DUR, AMP, RISE, AREA, AR/AM and SI of motor unit potentials (MUPs) were significantly lower than those of the controls (P < 0.05 or P < 0.01), especially AREA and SI (P < 0.01). But no remarkable difference was found between the muscles on the two sides of the diseased extremties. These findings suggest that quantitative analysis of MUPs is of importance to diagnosis of dermatomyositic disease and the parameters such as AREA and SI may provide more valuable information. PMID- 10684101 TI - [A longitudinal study on growth model and velocity of term small for gestation age]. AB - To probe into the vegetal pattern and find out the key period of promoting normal growth of term small for gestational age (TSGA), a longitudinal study on growth model and velocity of TSGA was conducted from Jan 1993 to June 1997. The body weight and length of 150 children of TSGA (57 boys, 93 girls) and 152 children as controls (58 boys, 94 girls) were measured from the 1st month to 36th month. The growth model was analysed by multilevel models. The result showed that there was a difference between the growth models of TSGA and controls. During 8 and a half months (center month), the weight, length of controls were 1.19 kg, 3.46 cm greater than those of TSGA; the weight, length of boys were 0.34 kg, 1.08 cm more than those of girls respectively. The growth velocity of TSGA was similar to that of controls. The maximal growth velocity was observed during the first 6 monthes after birth. The order of growth velocity from high to low was 1 mo. > 2 mo. > 3 mo. in length and weight, showing that children of TSGA have their own growth model. Their growth velocity should be monitored attentively. If low velocity appears one should search for the cause and adopt apropriate measures to ensure their growth in accordance with their "own track". PMID- 10684102 TI - [Relationship between PGI2, TXA2 and threatened preterm]. AB - The objectives of this study were to investigate the levels of 6-keto-PGF1 alpha, TXB2 and progesterone in plasma and cervical mucus of women with threatened preterm labor (Group I), term labor (Group III) versus their controls (Groups II and IV) respectively and to investigate the relationship between these substances and threatened preterm labor. One case was matched with one control (pregnant woman) by weeks of gestation. Radioimmunoassay was used to determine the levels of 6-keto-PGF1 alpha, TXB2 and progesterone. Statistical differences were assessed by student t test. The results showed that the levels of TXB2, 6-Keto PGF1 alpha in plasm from women with term labor and threatened preterm labor were higher than those of the control groups (P < 0.001) respectively. The ratios of 6 keto-PGF1 alpha and TXB2 were lower than their controls (P < 0.001) respectively. No differences in the levels of progesterone were noted between the labor groups and their control groups (P > 0.05) respectively. The results observed in cervical mucus were in line with there in plasma. These results suggest that the levels of PGI2, TXA2, and PGI2/TXA2 ratio were associated with the onset of term labor. PMID- 10684103 TI - [Effects of plasma endothelin-1 and aldosterone on sodium retention in children with nephrotic syndrome]. AB - The effects of plasma endothelin-1 (ET-1) and aldosterone (Ald) on sodium retention in children with nephrotic syndrome were investigated. 14 cases were divided into the edematous stage and non-edematous stage. Plasma ET-1, Ald, and serum osmolarity, albumin (Alb), and Na+ were measured in each stage. The results showed that the plasma ET-1 in edemaous stage increased significantly, compared with that in no-edematous stage (P < 0.01). There was no significant difference in plasma Ald between the edematous stage and the non-edematous stage. Plasma ET 1 was positively correlated with the serum sodium ionic concentration and negatively correlated with the serum Alb(r = 0.486, P < 0.01; r = 0.490, P < 0.01, respectively). In conclusion, the sodium retention with nephrotic syndrome might be correlated with reduction of secreted sodium in the kidney, suggesting that ET-1 plays an important role in pathogenesis. PMID- 10684104 TI - [Observations on rat's muscle at various postmortem intervals by scanning electron microscopy]. AB - The aim of this study was to observe the morphological changes of muscle in the process of rigor mortis. The quadriceps of 40 rats at various postmortem intervals were observed under the scanning electron microscope (SEM) and the light microscope by phosphtungstic acid-haematoxylin staining. The results showed that the striations of muscle were blurred within 4 hours, but they became apparent from 6 hours to 24 hours after death. The authors suggest that this phenomenon be associated with the increased resistance of muscle against the postmortal changes. The observations by scanning electron microscopy and light microscopy have revealed that the muscles do contract in the process of rigor mortis because the distance between two Z lines shortens and the I band narrows, compared with those in anaesthetised animals. The basic biochemical process for the formation of rigor mortis is the same as that of muscle contraction except that the former happens postmortem and the latter antemortem. PMID- 10684105 TI - [Sjogren's syndrome and malignant lymphoma]. AB - To diagnose and study the early stage of Sjogren's syndrome associated with malignant lymphoma, the authors used the PCR technique of B malignant lymphoma gene for lacrimal gland tissues in four patients with Sjogren's syndrome and in five patients suffering from B malignant lymphoma. The results showed that all cases of Sjogren's syndrome were negative and all cases of B malignant lymphoma were positive. These suggest that in general the patients with primary or secondary Sjogren's syndrome have no malignant lymphoma genes, but those who have chronic fever and enlarged superficial lymph nodes should under go this technique for early diagnosis and treatment of Sjogren's syndrome associated with malignant lymphoma. PMID- 10684106 TI - [Preparation of acyclovir-polybutylcyanoacrylate-nanoparticles by emulsion polymerization method]. AB - The aim of this study was to optimize the conditions and technology of preparing acyclovir-polybutylcyanoacrylate-nanoparticles (ACV-PBCA-NP) which has the diameter of about 100 nm and the shape of a sphere. The influential factors on sphericization were observed by single factor optimization. The preparation conditions and technology were optimized by the even design method. The contents of acyclovirin in acyclovir polybutyloganoacrylate nanoparticles were determined by HPLC. The optimum conditions and technology of preparing acyclovir polybutylcyanoacrylate nanoparticles were decided and put into use. The average diameter of the ACV-PBCA-NP thus prepared was 108.5 +/- 94.8 (n = 588). Its embedding ratio was 71.8%, and drug loading was 18.5%. The results suggest that the conditions and technology of preparing ACV-PBCA-NP presented in this paper are stable and practical. PMID- 10684107 TI - [Identification and determination of Shenbaiye]. AB - This study was amied at the method of identifying and determining Shenbaiye so that a ground can be provided for the quality requirements. Three Chinese materia medica (Radix Sophorae flavescentis, I; Cortex Phellodendri, II; Rhizoma Atractylodis, III) of Shenbaiye were identified by means of TLC successfully. The method in detail and the chromatogram were presented. The determination of I in Shenbiaye was carried out by solid phase cleaned-up and separated through neutral alumine column (10 x 1 cm i.d.) eluting with chloroform. This eluent was extracted with sulphuric acid (0.01 mol/L) and the excess of acids was titrated with sodium hydroxide (0.02 mol/L) using the methyl red as an indicator. The total alkaloid content of I was calculated as matrine of I in Shenbaiye. This method was established with a precision of 1.10% (n = 5) and the recoveries of 97.59 +/- 1.28% (n = 8). The results of determination of samples were reported. PMID- 10684108 TI - [Patch clamp recording in brain slice--modified blind method and the perforated patch clamp method]. AB - The use of the patch clamp recording in combination with the technique of the brain slice is the forefront method widely used in the studies of the neurocircuit in vitro. However, few authors in the papers published in domestic journals adopted this method. This paper presents the author's work to modify some details of blind method and the perforated patch clamp method. We used a dissecting microscope (with 40 times object lens) to replace the reverse microscope. As the recording electrode is very important in the experiment, we studied the size and the shape of the electrode tip and used two to three steps to pull the electrode so that the process of polishing of the electrode tip was not necessary and in perforated patch recording the synaptic transmission could be observed at the depolarization of 0-40 mV. The author also showed typical current or voltage waveforms in voltage or current mode respectively for guiding how to obtain the tight-seal whole cell patch. As these modifications have facilitated the acquisition of stable recording and lowered the cost of instrumentation, the methods become more suitable for the use in domestic laboratories. PMID- 10684109 TI - [CCD-diode array spectrophotometry used for simultaneous determination of cadmium and lead]. AB - A new spectrophotometric setup for simultaneous determination of lead and cadmium has been developed. It is composed of charge coupled device (array detector), multichromatic instrument and computer. The optimum chromogenic conditions of meso-tetra (4-trimethylammoniumphenyl) porphine (TAPP) with cadmium and lead were studied and reported in this paper. The linear ranges of Pb and Cd were 0-0.50 microgram/ml and 0-0.20 microgram/ml respectively. The detection limits were 0.001 microgram/ml for both Pb and Cd. The proposed method has been successfully applied to the simultaneous determination of cadmium and lead in the synthetic samples and the soaking solutions of ceramics and enamel. The recoveries of spiked samples ranged from 84.4%-118.6% and the average recovery was 100.8% with RSD of 1.0%-9.1%. The proposed method is more sensitive, more accurate and faster than the traditional one, it can be used to simultaneously determine multiple elements. PMID- 10684110 TI - [Gene expression and distribution in mouse abdominal cavity mediated by adenovirus]. AB - Infection and expression of recombinant human adenovirus in mouse abdominal cavity was reported. After adenovirus vector Ad/RSV-beta-gal harboring the E. coli lacZ marker gene was injected into mice abdominal cavity, the peritoneal surface of jejunum, ileum, colon, uterus, liver, spleen, stomach, bladder, abdominal wall, diaphragm and testis was found large patches of lacZ-positive cells. But the adenovirus vector was not able to penetrate the peritoneum, as demonstrated by histochemical staining. Another adenovirus vector Ad/RSV-tk harboring the HSV-tk gene was injected into mouse abdominal cavity and the mouse was treated with ACV. No acute toxic reaction was observed. Based on these data, the feasibility of gene therapy of malignant tumor within abdominal cavity with adenovirus mediated TK/GCV system was discussed. PMID- 10684111 TI - [Studies on cell signaling immunomodulated murine peritoneal suppressor macrophages: LPS and PMA mediate the activation of RAF-1, MAPK p44 and MAPK p42 and p38 MAPK]. AB - Monocytes-macrophages which serve as host immune cells to kill pathogens can often be "activated" after exposing to viruses, bacteria, cytokines as well as chemical substances, However, it is paradoxical that highly activated macrophages can be induced to become the suppressor ones by live microbes, microbial products, tumor, and autoimmune disease, although the mechanism remains unknown. Our previous experimental studies have shown that immuno-suppressor activities of suppressor macrophages on T, B and NK cells can be prevented by the treatment with LPS or supernatant in vitro from mitogen-stimulated lymphocytes, while, at the same time, the tumoricidal activities of those macrophages can be kept or even enhanced following the same treatment. This phenomenon was then termed as "immune modulation" For the understanding of its mechanism, we are now undertaking signal transduction in modulated macrophages. Since mitogen-activated protein kinase (MAPK) is an integration point of different signal transduction pathways, its cascade and regulation of activation are being investigated extensively by the assay of electrophoresis mobility shift. Recent results suggested that interaction of ligand-receptor triggers protein tyrosine kinase(PTK) activation leading to Ras-GTP binding with Raf-1 to phosphorylate MAPK kinase (MAPKK), the specific activator of MAPK. It is reported that PKC alpha can directly phosphorylate or activate Raf-1 in NIH3 T3 cells. Raf-1 (74 KDa), with an intrinsic serine (Ser)-threonine (The) kinase activity, becomes hyperphosphorylated after activation which can be followed by gel mobility shift test. It has also been shown that a variety of extracellular factors stimulate a pair of MAPK p44 and MAPK p42 of MAPK family members. A significant property of activation of ERK 1 and ERK 2 is the requirement for the phosphorylation of both Thr-183 and Tyr-185 (at TEY motif) within in its protein kinase subdomain VIII. More recently, two other MAPK subtypes, p38 MAPK (mammalian equivalents of HOG1 in yeast) and JNK MAPK have been discovered. The requirement for activation of p38 MAPK for both Thr-180 and Tyr-182 (at TGY motif) has been shown. p38 MAPK is important in certain transcriptional regulatory pathways, since it can phosphorylate the following transcriptional factors: 1) Elk at Ser 383/389 for binding with SRE motif; 2). ATF 2 at Ser 69/71, forming a complex with Myc for DNA binding at CRE motif; 3) Max at Ser-62 to combine DNA of E-Box motif. p38 MAPK can be activated by LPS, inflammatory cytokines, such as TNF and IL-1, osmolarity. To examine the possibility that whether activation of Raf-1 and ERK 1, ERK2 and p38 MAPK can be regulated directly or/and differently by PKC and PKA pathways, herbimycin A (Ki = 0.9 mumol/L), a potent PTK inhibitor (J. Immunol. 155:3944-4003, 1995) at 2 mumol/L concentration was utilized to block Ras/Raf 1/MAPK cascade. After pre-incubation of macrophages with herbimycin A for 30 min or 90 min, cells were treated with LPS (10 micrograms/ml) and PMA (100 nmol/L) for 15 min. No inhibition of phosphorylation of Raf-1, MAPK p44 and MAPK p42 in response to LPS and PMA was observed (Fig. 1 and 3). However, forskolin, a cAMP inducer for protein kinase A (PKA) activation, inhibited the phosphorylation of LPS- and PMA-stimulated Raf-1, MAPK p44 and MAPK p42 (Fig. 2 and 4). Similarly, in agreement with a very recent report from David, M et al in NIH, in which they indicated that forskolin (30 mumol/L) inhibited IFN-beta-stimulated ERK activity by U 266 cells (J. Biol. Chem. 271: 4585-4588 1996), we found that the levels of phosphorylations of Raf-1 and ERK1 and ERK2 were declined when forskolin (30 mumol/L) was added to macrophages for 20 min at 37 degrees C prior to the stimulation by LPS and PMA. Interestingly, under the same condition, forskolin (30 mumol/L) stimulated the phosphorylation of LPS- and PMA-triggered p38 MAPK of murine peritoneal suppressor macrophages, suggesting that activatio PMID- 10684112 TI - [The expression of Quox-1 gene homologous sequence in the development of early human embryos]. AB - By using the b2 fragment of Quox-1 gene as probe, we have confirmed that the Quox 1 gene homologous sequence exists in the human genome according to the results of Southern blot. Studies on the expression of Quox-1 homologous sequence in early human embryos from 26 to 37 days by means of immunohistochemistry technigue with Quox-1 protein antibodies showed the spatiotemporal expression patterns: in 26 days embryo Quox-1 homologous sequence was expressed in many places including neural tube, but 30 days later, tits expression sites were limited to notochord, digestive epithelium, myotome, cardiac muscle cell and periderm. The functions in control and regulation of Quox-1 gene homologous sequence during the early development of human embryo were discussed. PMID- 10684113 TI - [Spatial and temporally expressed proteins during Drosophila oogenesis as revealed by monoclonal antibodies]. AB - By use of hybridoma technique, we have prepared 6 monoclonal antibodies. As shown by antibody labeling of whole mount ovary, four of them recognized, respectively, the antigens which were spatial-temporally expressed during oogenesis. The B 2 antigens appeared very early in the germarium and were expressed mainly by cystocyte and nurse cells. Later, they were all transported and localized in the posterior of oocyte, F 9 antigens followed and were also transported and localized in the posterior of oocyte in the stage 7-8. Then, E 8 antigens appeared and mainly localized on the membrane of oocyte in the stage 9-10. C 3 antigens were expressed much later, at about stage 14. They localized like two caps in the perivitelline fluid at both ends of matured egg. Such specific expression and distribution pattern of all these antigens suggest their possible roles during oogenesis. PMID- 10684114 TI - [Regulation of human 17 beta-hydroxysteroid dehydrogenase type 1 expression by cyclic adenosine 3',5'-monophosphate in choriocarcinoma cells]. AB - Human 17 beta-hydroxysteroid dehydrogenase type 1 (17 HSD type 1), encoded by HSD 17 B 1 gene, is a steroidogenic enzyme catalyzing the interconversion of estrone and estradiol. In this study, we investigated the role of cyclic adenosine 3', 5' monophosphate (cAMP) in the regulation of 17 HSD type 1 expression in cultured choriocarcinoma cell lines. Treatment with 8-bromo-cAMP increased 17-HSD type 1 protein concentration in JAR and JEG-3 cells, and the induction was accompanied by parallel increase of 1.3 kb 17 HSD type 1 mRNA expression. Reporter gene analysis revealed that the activity of HSD 17 B 1 promoter in JAR and JEG-3 cells was induced by cAMP and that the region participating in transmission of cAMP effect is situated in the position between -659 and -550 in HSD 17 B 1 gene. The consequent electrophoretic mobility shift assay showed that this region formed specific DNA-protein complexes with nuclear extracts prepared from JAR, JEG-3, T 47 D and HeLa cells. The data provide the first evidence that HSD 17 B 1 gene transcription is activated by cAMP in choriocarcinoma cells. PMID- 10684115 TI - [The effects of hydroxyurea on cell-cycle distribution and the expression of human beta-globin gene in K 562 cells]. AB - Although the hydroxyurea (HU) has been extensively studied, little is known of its molecular mechanism in controlling the expression of human globin gene and in modulating the progression of cell-cycle in K 562 cell. In the present study, the effect of hydroxyurea on proliferative kinetics of K 562 cells was examined by monitoring the number of cells during a period of 8 day's cell culture. Our results showed that there was a dose related decrease in cell growth when K562 cells were incubated with HU. Moreover, cell-cycle analysis demonstrated that HU had profound effect on cell-cycle distribution. In the case of the induced K 562 cells, there was an increased accumulation of cells in S phase and a decreased fraction of cells in G 1 and G 2 + M phase. Furthermore, HU could induce the expression of human beta-globin gene in the induced K 562 cells. Our results indicate that HU has a potential to inhibit the proliferation of K 562 cells and to stimulate the terminal differentiation of this cell. PMID- 10684116 TI - [Study on the transformation of mammal cells induced by glycidyl methacrylate in vitro]. AB - Glycidyl methacrylate(GMA) can induce the phenotype transformation of human embryonic lung fibroblasts. The transformed cells prolifeated rapidly with a higher in the ratio of nucleus to cytoplasm, and their life span extended notably. The transformed colonies exhibited in extensive random orientation and the cells crossing-over. The transformed cells could be agglutinated by lower concentration of ConA and could grow in semisolid agar. The chromosome abberations could occur in transformed fibroblasts. These results suggest that GMA is able to induce malignant transformation of mammal cells, and it may be carcinogenic to human. PMID- 10684117 TI - [Immobilization of rat liver microsomes with liquid lipid membrane encapsulation]. AB - Rat liver microsomes were immobilized by encapsulating them into liquid lipid membranes (56% paraffin thick, 38% paraffin thin, 5% lecthin and 1% dodecanol). The resistance against heat and pH profile of the immobilized enzyme-NADPH cytochrome C reductase was weakened. However, the activity and storage stability of the immobilized enzyme were enhanced. PMID- 10684118 TI - [Effects of propyl gallate on histopathology of liver from mice exposed to trinitrotoluene]. AB - In addition to the hepatocellular edema and cytoplasmic eosinophilia, sludging of blood was present in liver of mice exposed to trinitrotoluene(TNT). Single necrosis of the partical liver cell was seen occasionally. Liver damage induced by TNT was significantly alleviated by orally administrated propyl gallate(PG). Futhermore, PG can promote the regeneration of the hepatocytes following TNT exposed mice. The results suggest that PG showed a protective effect on the histopathologic changes of liver injury induced by TNT. PMID- 10684119 TI - [Risk of lung cancer among iron and steel workers in Anshan, China--case-control study]. AB - A case-control interview study on 610 lung cancer patients and 959 controls was conducted among male workers in Anshan Iron-steel Complex. After adjusting for non-occupational risk factors, such as smoking, pulmonary disease, family history of cancer and the consumption of fruit, risks for lung cancer were significantly higher in workers engaged in smelting and rolling (OR = 1.5, 95% CI = 1.1-2.2), in the fire-resistant brick factory (OR = 2.9, 95% CI = 1.4-5.9), in general loading (OR = 2.5 95% CI = 1.0-6.1), and in coking (OR = 3.4, 95% CI = 1.4-8.5) for 15 or more years. Significant dose-response was observed for exposure to total dust and B[a]P, but not for specific chemical components of dust. The lung cancer risk increased 40% in iron and steel workers with long term occupational exposure. PMID- 10684120 TI - [Study on the health of cotton mill workers]. AB - A group of 110 workers in a cotton mill was investigated by questionnaire, skin testing, and the measurement of airway responsiveness through forced expiratory volume for one second(FEV1) by spirometry. The workers were examined before starting work, 10 weeks and one year after exposure. Decreases in FEV1 over shifts were small at 10 weeks and one year, and were slightly higher among people with positive skin reactions to cotton dust extracts. Airway responsiveness, defined as the average decrease in FEV1 after 1.25 mg methacholin, was increased at 10 weeks. It remained about the same after one year, except in the workers with positive skin test. Subjective symptoms of chest tightness and cough with phlegm increased progressively at 10 weeks and one year. Nasal irritation remained unchanged and dry cough decreased in one year. The results suggest that the airway inflammation caused by cotton dust increases with the exposure time and that the changes are more notable in workers with reactivity to cotton dust extract. PMID- 10684122 TI - [Effect of particles size calibration on air microbe sampling]. AB - The airborne bacterial particles number sampled with 6 grades Andersen sampler at Xidan in Beijing from 1987 to 1988 was calibrated by the alive bioparticles calibration formula. The results showed that the average number of airborne bacterial particles sampled in 3 minutes and in 84.9 L air was 257, but it is 315 when calibrated. The later was higher than the former (t = 2.012, P < 0.05). The average number sampled with the 1st grade sampler (> 8.2 microns) was 98, but it is 136 when calibrated, and the difference was obviously (t = 2.409, P < 0.05). There was no obvious difference between the number sampled with the rest grades sampler (2nd-6th) and when calibrated (t = 1.701-0.026, P > 0.05). The effects of calibration on the particles size distribution and its concentration day's change were not obviously. PMID- 10684121 TI - [Influence of arsenic on proliferation and differentiation of rat bud cells in vitro]. AB - The objective of this study was designed to evaluate the developmental toxicity of arsenic and its effect on embryonic chondrogenesis of Sprague-Dawley rat by using Flint's rat limb bud cell micromass cultrure system in vitro. The results revealed that arsenic inhibited markedly both proliferation and differentiation of rat limb bud cells in vitro and there was an obvious dose-response relationship. The concentrations of arsenic for IP50(dose of inhibiting proliferation by 50% of the control value) and ID50(dose of inhibiting differentiation by 50% of the control value) were 0.70 mg/L and 0.21 mg/L respectively. The ratio between IP50 and ID50 was 3.3. These parameters indicated that the influence of arsenic on differentiation of rat limb bud cells was stronger than on proliferation, and arsenic belonged to a strong teratogenic agent and a specific inhibitor. This study suggested that the specific inhibitory action of arsenic on limb bud cell differentiation did not result from the cytotoxicity, but would result from the teratogenic effects of arsenic. PMID- 10684123 TI - [Development of emission models for volatile organic compounds from indoor materials]. AB - Volatile organic compounds (VOCs) emitted from indoor materials was a major cause of indoor air pollution. The characteristics of VOCs emission was an important part of research programs on indoor air quality. The technology of test chambers with exactly controllable conditions has been successfully used in studies of VOCs emissions. The technology could be used to model the chamber VOCs concentration level vs time profile C(t), which could in turn be used to estimate the sample emission rate vs time profile R(t). The emission models of VOCs from indoor materials were presented in this review. The principal of emission process, parameters and the applications of emission models were introduced. The application of diffusion model, dilution model and vapor pressure (VP) model were limited due to the existence of sink effect. Sink model is the most promising model at present. PMID- 10684124 TI - [Study on the preventive effect of tea on DNA damage of the buccal mucosa cells in oral leukoplakias induce by cigarette smoking]. AB - In order to study the preventive effect of tea on DNA damage induced by cigarette smoking and to provide further evidence on the protective effects of tea in human cancer, a 6-month, double-blind and randomized placebo-controlled intervention trial was carried out. The effect of tea on micronuclei frequency in exfoliated oral buccal mucosa cells in 36 oral leukoplakias of smokers was investigated. A kind of mixed tea given in an oral dose of 3 g/day and a concentration of 0.1% smeared on mucosa lession three times a day for 3 months and 6 months, significantly decreased the micronuclei formation in exfoliated oral buccal mucosa cells in subjects with oral leukoplakias. In contrast, there was no change in the micronuclei frequency after 6 months in the placebo group. The results indicate that mixed tea may reduce the oral cancer risk by preventing DNA damage in oral leukopiakias induced by cigarette smoking. PMID- 10684125 TI - [Study on the relative absorption of food carbohydrate in healthy adults]. AB - The absorption of food carbohydrate was determined with breath hydrogen test (BHT) in healthy adults. When the absorption of lactulose was assumed as 0, the absorption of food carbohydrates were as follows: 100% for rice with ground lean pork; 98% for rice; 85% for bread; 58% for corn and 30% for red potato. The absorption of food carbohydrate was positively related to the oral-colon transit time (OCTT, r = 0.8790) and the peak hydrogen time (PHT, r = 0.6745), and was negatively related to the peak hydrogen value (PHV, r = -0.6468) and to the hydrogen producing percentage (H2 > 1.8 mg/m3, r = -0.8679). The breath hydrogen test can be used in estimating the absorption of food carbohydrate in human body because this method is safe, non-invasive, reproducible and accurate. PMID- 10684126 TI - [Effects of high dietary zinc on liver function, hepatic drug metabolism enzymes and membrane fluidity in mice]. AB - Mice were fed with high zinc diet (15 g/kg) for 3 weeks. High zinc could cause liver toxicity: 1. inhibiting the activity of GOT and GPT in liver homogenate, reducing GSH and glycogen contents. 2. increasing the activity of aniline hydroxylase and inhibiting the activities of NADPH-cytochrome C reducease, benzo phytamine-N-demethylase and glutathione S-transferase. The activities of cytochrome P450 and cytochrome b5 were not obviously changed 3. increasing microsomal membrane fluidity in the superficial layers, but not in the deep layers. PMID- 10684127 TI - [Protection of vitamin E against testis lipid peroxidation induced by iron and ethanol]. AB - The influence of acute iron and ethanol load on lipid peroxidation and antioxidative defense systems in rat testes and the modification after supplemented with Vitamin E was investigated. Acute iron and ethanol load was achieved by i.p. injection of iron-dextran (500 mg/kg) and ethanol (50 mmol/kg). After 18 h, a significant increase in testis total iron content was induced. Compared with control, total testis iron content was 6.8-fold higher in iron treated rats and 9.1-fold in iron and ethanol treated rats. As the content of iron increasing, the endogenous lipid peroxidation evaluated as 2-thiobarbituric acid-rective substances (TBARS) increased apparently, and the content of lipid soluble antioxidants alpha-tocopherol decreased. The supplement of an antioxidant, alpha-tocopherol, protected against lipid peroxidation. Iron and ethanol treatment did not affect the activity of superoxide dismutase, catalase and glutathione peroxidase. The results indicate that acute iron load causes iron accumulation in rat testes and ethanol increases the same accumulation. Iron played pivotal paracrine roles on ethanolinduced injure rat testis. The supplement of Vitamin E can protect against this damage. PMID- 10684128 TI - [Study on the human dietary intake of minerals, energy, fiber and phytic acid]. AB - Duplicate diets of three days were collected. The diets represented general living standard of male adults. There were three types of subjects: city inhabitants in group 1, northern peasants in group 2, and southern peasants in group 3. The diets were analyzed to calculate the average per capita intake of various minerals, energy, fiber and phytic acid. The assessment of intake according to the values of RDA, ESADDI and ADI shows the following results: Zn is low, Ca comes up to 2/3 of RDA, P reaches 88%-133% of RDA, Fe appears enough to attain the standard, but the availability should be considered. The contents of Se and I in each group was varied. Even though the intake of Se is low, it reaches the minimum requirement. Since I is being supplemented for all the people, it would not be deficient. The intake levels of Cu, Mo and Cr achieve or surpass the minimum values of ESADDI. Mn intake exceeds the maximum value of ESADDI, but the highest intake was only 10. 5 mg, which is far from poisoning level. High Na and low K intakes are specific defects of Chinese diets. The level of Cd, Hg and Pb is far apart from toxicosis. As for the amount of As intake, only in group 3 is higher than ADI by 29%. The energy is full. Dietary fiber is suitable. The intake of phytic acid of northern peasants is higher than that of southern peasants and city inhabitants, but it would be harmless. PMID- 10684129 TI - [Determination of taurine in food by high performance liquid chromatography]. AB - Taurine in food was separated by HPLC as its phthalaldehyde-ethanethiol derivative on mu-Bondapak C18 reverse column and measured at 330 nm. The determination was not interfered by sulphosalicylic acid, excess dervatizing reagents and 20 kinds of amino acids. The stability of the taurine dervatives was discussed in this paper. The result showed that the higher concentration of borate buffer, the more stable of taurine derivatives and the lower concentration of taurine, the faster decomposition of taurine dervitives. Using more concentrated borate buffer (0.4 mol/L) and controlling the time between injection and reaction and keeping the operation identically could avoid the effect of nonstable taurine derivatives in the determination. The minimum detectable quantlity was 8 ng. The coefficient of variation was less than 8%. The recoveries were 90.7%-105.1%. PMID- 10684130 TI - [Simultaneous extraction of tea-polyphenols and caffeine from green tea]. AB - Tea-polyphenols and caffeine were extracted simultaneously from green tea. The factors influencing on the process of impregnation and extraction were studied. The result indicated that the content of tea-polyphenols and caffeine in tea was increased with the duration of extraction and decreased with the frequency of extraction. The authors discuss the effect of pH on the precipition of calcium tea-polyphenols. PMID- 10684131 TI - [Simultaneous determination of lead and copper in food by differential potential stripping with the presence of emulgent OP]. AB - The effect of different surfactants on the stripping analysis of Pb and Cu was studied. The optimum condition for simultaneous determination of Pb and Cu in foods by the differentital potential stripping analysis in the 0.3 mol/L HCl media and by using emulgent OP as spike agent was described. The contents of Pb and Cu in soft drinks, fermented wine and flavourings with added standards were determined directly by synchronous mercury plating. The recovery of the added standard samples were 95%-105%, and relative standard deviation was 0.8%-7.9%. The proposed method has been applied to the sample analysis. PMID- 10684132 TI - [Study on the antagonistic action of selenite on fluoride-induced lipid peroxidation and on the changes of trace elements in rats]. AB - Five groups of SD male rats were provided with deionized drinking water containing 0 and 150 mg/L NaF, and containing both 150 mg/L NaF and 0.5, 2.0 or 4.0 mg/L Na2SeO3 respectively for 10 weeks, in order to find out the optimal level of selenite in drinking water against fluoride toxicity. The results showed that fluoride could cause significant increase of lipid peroxides (LPO) and metabolic disorder of trace elements in the serum and kidney of rats. The antagonistic effect of 2.0 mg/L Na2SeO3 drinking water on the lipid peroxidation induced by 150 mg/LNaF was the most evident, whereas those of 0.5 and 4.0 mg/L Na2SeO3 were not obvious. It is concluded that selenite possesses significant antagonistic effects on renal damages induced by fluoride and 2.0 mg/L Na2SeO3 is the optimal concentration for the antagonistic effect on renal impairments induced by 150 mg/L NaF in drinking water. PMID- 10684133 TI - [Study on the joint action of selenium and cadmium on DNA damage of rat liver cells]. AB - The effect of joint action of selenium and cadmium on DNA damage in rat liver cells was investigated with single cell gel electrophoresis. The results show that both selenium and cadmium can induce DNA damage at the concentration of 8.75 mumol/L, 17.5 mumol/L and 35 mumol/L. The degree of DNA damage induced by cadmium is more serious than by selenium. When selenium and cadmium act jointly, they can antagonize each other on DNA damage at the concentration of 8.75 mumol/L and 17.5 mumol/L, but not at the concentration of 35 mumol/L. There is an antagonism between selenium and cadmium on DNA damage in rat liver cells at certain concentrations. PMID- 10684134 TI - [Effect of dietary selenium on the activities of glutathione peroxidase and deiodinase in rat liver]. AB - In order to investigate the effect of dietary selenium on the activities of glutathione peroxidase and deiodinase and the least selenium requirement for their optimal activities, fifty four male weanling Wistar rats of 50-60g body weight were randomly divided into 9 groups and were fed with semisynthetic diets of different selenium level for 20 weeks. The selenium in the diets were 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.1, 0.2 and 5 mg/kg diet respectively. The body weight of ratsin the ninth group was different significantly from the seventh and eighth group, but those in the other eight groups were not significantly different. The activities of liver glutathione peroxidase in 0.1, 0.2 and 5 mg/kg groups were the highest among the nine groups. So at least 0.1 mg/kg diet is required for its normal activity. The activities of the liver type I deiodinase of nine groups were high from 0.05 to 0.2 mg/kg diet groups and at least Se 0.05 mg/kg diet is required for its normal activity. PMID- 10684135 TI - [Calculating method for the necessary lamps and sterile rate in a tube-shaped ultraviolet air washer]. AB - It has much more advantage to use the cylindric ultraviolet air washer than to use the ordinary ultraviolet lamps. There was a calculation method for determining necessary lamps in a rectangled ultraviolet air washer, but it had a limiting condition. This paper developed two calculating methods for determining necessary lamps and its sterile rate in a tube-shaped ultraviolet air washer. The sterile rate can be extracted with any parameter. Necessary lamps can also be extracted with its sterile rate. PMID- 10684136 TI - 28th Annual International Neuropsychological Society Conference. Denver, Colorado, USA. February 9-12, 2000. Abstracts. PMID- 10684137 TI - Assessment of functional capacity through oxygen consumption in patients with asymptomatic probable heart disease. AB - PURPOSE: To compare peak exercise oxygen consumption (VO2peak) of healthy individuals with asymptomatic individuals with probable heart disease. METHODS: Ninety-eight men were evaluated. They were divided into two groups: 1) 39 healthy individuals (group N) with an age range of 50 +/- 4.6 years; and 2) 59 asymptomatic individuals with signs of atherosclerotic and/or hypertensive heart disease (group C) with an age range of 51.9 +/- 10.4 years. In regard to age, height, body surface area, percentage of fat, lean body mass, and daily physical activity, both groups were statistically similar. Environmental conditions during the ergometric test were also controlled. RESULTS: Maximal aerobic power (watts), VO2peak, maximal heart rate, and maximal pulmonary ventilation were lower in group C (p < 0.01) than in group N; weight, however, was lower in group N (p = 0.031) than in group C. Differences in the respiratory gas exchange index, heart rate at rest, and the maximal double product of the two groups were not statistically significant. CONCLUSION: Signs of probable heart disease, even though asymptomatic, may reduce the functional capacity, perhaps due to the lower maximal cardiac output and/or muscle metabolic changes. PMID- 10684138 TI - Coronary artery disease, microalbuminuria and lipid profile in patients with non insulin dependent diabetes mellitus. AB - PURPOSE: To determine the frequency of coronary artery disease, microalbuminuria and the relation to lipid profile disorders, blood pressure and clinical and metabolic features. METHODS: Fifty-five type 2 diabetic patients (32 females, 23 males), aged 59.9 +/- 9 years and with known diabetes duration of 11 +/- 7.3 years were studied. Coronary artery disease (CAD) was defined as a positive history of myocardial infarction, typical angina, myocardial revascularization or a positive stress testing. Microalbuminuria was defined when two out of three overnight urine samples had a urinary albumin excretion ranging 20-200 micrograms/min. RESULTS: CAD was present in 24 patients (43.6%). High blood pressure (HBP) present in 32 patients (58.2%) and was more frequent in CAD group (p = 0.05) HBP. Increased the risk of CAD 3.7 times (CI[1.14-12]). Microalbuminuria was present in 25 patients (45.5%) and tended to associate with higher systolic blood pressure (SBP) (p = 0.06), presence of hypertension (p = 0.06) and known diabetes duration (p = 0.08). In the stepwise multiple logistic regression the systolic blood pressure was the only variable that influenced UAE (r = 0.39, r2 = 0.14, p = 0.01). The hypertensive patients had higher cholesterol levels (p = 0.04). CONCLUSION: In our sample the frequency of microalbuminuria, hypertension, hypercholesterolemia and CHD was high. Since diabetes is an independent risk factor for cardiovascular disease, the association of others risk factors suggest the need for an intensive therapeutic intervention in primary and in secundary prevention. PMID- 10684139 TI - Safety and efficacy of coronary stent implantation. Acute and six month outcomes of 1,126 consecutive patients treated in 1996 and 1997. AB - PURPOSE: The authors analyzed the 30-day and 6-month outcomes of 1,126 consecutive patients who underwent coronary stent implantation in 1996 and 1997. METHODS: The 30-day results and 6-month angiographic follow-up were analyzed in patients treated with coronary stents in 1996 and 1997. All patients underwent coronary stenting with high-pressure implantation (> 12 atm) and antiplatelet drug regimen (aspirin plus ticlopidine). RESULTS: During the study period, 1,390 coronary stents were implanted in 1,200 vessels of 1,126 patients; 477 patients were treated in the year 1996 and 649 in 1997. The number of percutaneous procedures performed using stents increased significantly in 1997 compared to 1996 (64% vs 48%, p = 0.0001). The 30-day results were similar in both years: the success and stent thrombosis rates were equal (97% and 0.8%, respectively). The occurrence of new Q wave MI (1.3% vs 1.1%, 1996 vs 1997, p = NS), emergency coronary bypass surgery (1% vs 0.6%, 1996 vs 1997, p = NS) and 30-day death rates (0.2% vs 0.5%, 1996 vs 1997, p = NS) were similar. The 6-month restenosis rate was 25% in 1996 and 27% in 1997 (p = NS); the target vessel revascularization rate was 15% in 1996 and 16% in 1997 (p = NS). CONCLUSIONS: Intracoronary stenting showed a high success rate and a low incidence of 30-day occurrence of new major coronary events in both periods, despite the greater angiographic complexity of the patients treated with in 1997. These adverse variables did not have a negative influence at the 6-month clinical and angiographic follow-up, with similar rates of restenosis and ischemia-driven target lesion revascularization rates. PMID- 10684140 TI - Influence of the elevation of the left ventricular diastolic pressure on the values of the first temporal derivative of the ventricular pressure (dP/dt). AB - PURPOSE: To assess the effects of the elevation of the left-ventricular end diastolic pressure (LVEDP) on the value of the 1st temporal derivative of the ventricular pressure (dP/dt). METHODS: Nineteen anesthetized dogs were studied. The dogs were mechanically ventilated and underwent thoracotomy with parasympathetic nervous system block. The LVEDP was controlled with the use of a perfusion circuit connected to the left atrium and adjusted to the height of a reservoir. The elevation of the LVEDP was achieved by a sudden increase in the height of a reservoir filled with blood. Continuous recordings of the electrocardiogram, the aortic and ventricular pressures and the dP/dt were performed. RESULTS: Elevation of the LVEDP did not result in any variation of the heart rate (167 +/- 16.0 bpm, before the procedure; 167 +/- 15.5 bpm, after the procedure). All the other variables assessed, including systolic blood pressure (128 +/- 18.3 mmHg and 150 +/- 21.5 mmHg), diastolic blood pressure (98 +/- 16.9 mmHg and 115 +/- 19.8 mmHg), LVEDP (5.5 +/- 2.49 and 9.3 +/- 3.60 mmHg), and dP/dt (4,855 +/- 1,082 mmHg/s and 5,149 +/- 1,242 mmHg/s) showed significant increases following the expansion of the ventricular cavity. Although the elevation of the dP/dt was statistically significant, 6 dogs curiously showed a decrease in the values of dP/dt. CONCLUSION: Sudden elevation of the LVEDP resulted in increased values of dP/dt; however, in some dogs, this response was not uniform. PMID- 10684141 TI - Natural course of subsequent pregnancy after peripartum cardiomyopathy. AB - OBJECTIVE: To assess the effect of subsequent pregnancy after peripartum cardiomyopathy (PPCM) on maternal and fetal outcome. METHODS: Prospective study of 34 patients with the diagnosis of PPCM (mean age = 26 years). At the time of first diagnosis 5 were in NYHA functional class (FC) II for heart failure, one in FC III and 28 in FC IV. After clinical treatment, patients were advised to avoid new pregnancies and a follow-up was obtained. RESULTS: There were 12 (35.3%) subsequent pregnancies in patients (pt) aged 19 to 44 years (mean 32), divided into two groups: GI: 6 pts who had normalized their heart size and GII: 6 pts with persistent cardiomegaly. GI had initially mild clinical manifestations (3 were in FC II, 1 in FC II and 2 in FC IV) and complete recovery of cardiac function (FC I). A new pregnancy was well-tolerated in 5 (83.3%); 1 pt presented with preeclampsia, and progressed to FC II. Presently, 5 pt are in FC I and 1 in FC II. GII pts had more severe heart failure at the onset of PPCM (1 pt in FC II and 5 in FC IV); during follow-up, 4 pt were in FC I and 2 in FC II. A new pregnancy was well tolerated in all of them, but the eldest, who had had 2 pregnancies and had a progressive worsening of clinical status, dying 8 years after the last pregnancy and 13 years after the diagnosis of PPCM. The remaining 5 pt are still alive, 3 in FC I and 2 in FC II, with worsening of FC in 1. Subsequent pregnancies occurred 3-7 years after clinical treatment of PPCM and no fetal distress was observed. CONCLUSION: Subsequent pregnancies are well tolerated after PPCM, but not devoid of risk. No fetal distress was observed. A minimum interval of 3 years after the recovery of function seems to be safe for subsequent pregnancies. PMID- 10684142 TI - Aortopulmonary window. Clinical and surgical assessment of 18 cases. AB - OBJECTIVE: Aortopulmonary window (APW) is an uncommon congenital malformation. Its clinical presentation is dependent on the size of the defect and on the associated lesions. We evaluated our experience with this anomaly and compared it with 296 cases reported in the literature. METHODS: Retrospective study of 18 patients diagnosed as having APW (age range from 13 days to 31 years, 13 (72.2%) females), divided into two groups: Group A (GA): 10 patients with isolated APW, and Group B (GB): 8 patients with associated lesions. RESULTS: Heart failure occurred in 14 patients, and cyanosis in 3:2 from GB (tetralogy of Fallot--TF, and double outlet right ventricle--DORV), and one from GA with pulmonary hypertension. In 5 patients from GA the diagnosis of mitral regurgitation was made based on a systolic murmur and LV hypertrophy on the EKG. In GB, clinical findings were determined by the associated defect. Diagnosis was established by echocardiography in 11 (61.2%) of the patients. In 3 patients, a wrong diagnosis of mitral regurgitation was made, in 1 a patent ductus arteriosus was diagnosed and in 3 others, the diagnosis of APW was masked by other important associated defects (2 cases of DORV and 1 case of TF). The diagnosis was made by catheterization in 3 (16.6%) patients, by surgery in 3 (16.6%) and by necropsy in 1 (5.5%). Corrective surgery was performed in 14 (77.7%) patients, with one immediate death and good long-term follow-up in the remaining patients. CONCLUSION: APW can be confused with other defects. Clinical findings, associated with an adequate echocardiogram can provide the information for the correct diagnosis. PMID- 10684143 TI - Aortic valve assessment. Anatomical study of 100 healthy human hearts. AB - PURPOSE: To assess anatomical characteristics of the aortic valve, so that they may be useful in diagnostic situations and surgical treatment. METHODS: The study analyzed 100 healthy fixed human hearts; 84% of them obtained from males, 61% of them from Caucasian individuals. The ages of the individuals ranged from 9 to 86 years (mean 30 +/- 15.5 years). The characteristics assessed related to age, sex, and race were the following: number and height of the cusps, size of the lunulae, internal and external intercommissural distance, position of the coronary ostium in relation to the aortic valve, position of the ventricular septum in relation to the aortic valve, thickness of the cusps. RESULTS: All hearts assessed had a tricuspidal aortic valve. In regard to the height of the cusps and size of the lunula, the left coronary cusp was larger, followed by the right coronary cusp and the noncoronary cusp. The internal and external intercommissural distances had mean values of 24.6 +/- 5.7 mm and 19.7 +/- 7 mm, respectively. In regard to the position of the coronary ostia, in one heart two ostia emerged from the left coronary sinus, and in another, the ostium was supracommissural. The mean diameter of the aorta was 21.8 +/- 3.6 mm, and there were no significant sexual or racial differences, but the diameter increased progressively with the increase in age. The thickness of the cusps did not show any significant difference in the 3 points assessed. CONCLUSION: The aortic valve annulus did not show a perfect circumference, with some variations in the measurements of the annulus, in the cusps and in the relation with the ventricular septum. PMID- 10684144 TI - Myocardial repair with long-term and low-dose administration of a nitric oxide synthesis inhibitor. Myofibroblasts, type III collagen and fibronectin. AB - OBJECTIVE: To study the healing process of the myocardium in hypertensive rats undergoing inhibition of nitric oxide synthesis. METHODS: Two groups of animals were studied: one received L-NAME, 12 mg/kg/day, and the other was a control group. The presence of type III collagen, fibronectin, and alpha-smooth muscle actin-positive cells was assessed by immunohistochemistry. RESULTS: Fibronectin was seen in both early and late lesions, while type III collagen was seen mainly in areas of incomplete healing, situated among myocytes and around the intramyocardial branches of the coronary arteries. Areas representing early and late lesions showed a population of spindle-shaped cells. Immunohistochemistry showed that these cells were positive for alpha-smooth muscle actin. CONCLUSION: In the myocardium of hypertensive rats, the alpha-smooth muscle actin-positive cells are related to the accumulation of type III collagen and fibronectin in the areas of myocardial damage. PMID- 10684145 TI - Essential thrombocythemia and acute myocardial infarction treated with rescue coronary angioplasty. AB - A 48-year-old man with essential thrombocythemia suffered an extensive anterior acute myocardial infarction; this is a rare association. A pharmacological thrombolysis was performed, without success. He subsequently underwent successful rescue coronary angioplasty. To our knowledge, there is no other report in the literature relating the triad of essential thrombocythemia, acute myocardial infarction and rescue coronary angioplasty. PMID- 10684146 TI - Left ventricular hypertrophy in systemic hypertension. Benefits of its reversal. PMID- 10684147 TI - Physical activity at moderate and high altitudes. Cardiovascular and respiratory morbidity. PMID- 10684148 TI - [Banco Mundial/FNUAP/OMS/PNUD. Special program of investigation, development and formation of investigators on human reproduction. Guidelines for the creation of scientific and ethical revision agencies]. PMID- 10684149 TI - [Ethical conflicts in the exercise of the role of health agents]. PMID- 10684150 TI - [Lanari Lecture. Hematopoietic cytokines and hematologic disorders]. PMID- 10684151 TI - Current trends in antiretroviral therapy for HIV infection and AIDS. PMID- 10684152 TI - [Studies of drug utilization: a necessary instrument to promote the rational use of drugs]. PMID- 10684153 TI - The steroidogenic acute regulatory (StAR) protein. PMID- 10684154 TI - Steroidogenic factor 1: a key mediator of endocrine development and function. PMID- 10684155 TI - Sources and function of neuronal signalling molecules in the gonads. AB - While the hypothalamic-pituitary-gonadal axis is crucial for the function of the gonads, non-endocrine regulatory influences are exerted by other factors within the gonads. Among these factors are neurotransmitters, such as catecholamines. Several types of receptors for catecholamines exist in the gonads on vascular or endocrine cells. Their activation can alter blood flow, steroidogenesis and gene expression, depending on the target cells. Recently a neuronal-like cell type expressing catecholamine-biosynthetic enzymes and neuronal proteins was identified in testis and ovary of human and non-human primates. Together with the well-known sympathetic innervation, this gonadal nervous system may serve as a source of catecholamines. Dopamine is present in the follicular fluid. Oocytes, while not able to perform de novo synthesis of catecholamines, were shown to utilize dopamine to produce norepinephrine. This catecholamine then acts on beta adrenoreceptors of follicular cells to increase cAMP. Oocytes may thus indirectly via dopamine and cAMP be able to control their own meiotic arrest. In addition, neurotransmitters may also be synthesized in other, non-neuronal ovarian cells. Thus, cultured human granulosa-luteal cells possess the acetylcholine synthesizing enzyme and the acetylcholine-specific vesicular transporter protein. These cells also express muscarinic-receptors (M1), which are linked to the mobilization of intracellular calcium and cell proliferation. This suggests involvement of the acetylcholine system in follicular growth and in the periovulatory events. In neurons, neurotransmitters alter the properties of the neuronal cell membrane. If this is the case in endocrine cells of the gonads is not yet clear, but the recent identification of voltage-activated potassium and sodium channels in human luteinized granulosa-luteal cells raises this question and opens a door to a new area of investigation. PMID- 10684156 TI - Glypican-3 is a novel inhibitor of insulin-like growth factor signaling. PMID- 10684157 TI - The role of plasminogen activator receptor in cancer invasion and dormancy. AB - Urokinase plasminogen activator receptor (uPAR) has been identified some 15 years ago and the anticipation was that its presence on the cell surface will provide a focus for anchoring uPA and possibly protect the enzyme from native inhibitors. The studies of the last decade have shown that uPA localized to the surface of cells by uPAR is indeed an important factor in the process of cancer cell invasion and metastasis. We developed a chick embryo model in which we showed that uPAR is crucial in invasion of stroma and in intravasation (breaching of the blood vessels walls). More recently and unexpectedly, uPAR--a protein anchored in the outer leaf-let of the plasma membrane, has been shown to initiate signal transduction events and affect cell migration. We have shown that uPAR co associates with fibronectin binding integrin, alpha 5 beta 1, activates them and that this interaction leads to a greatly increased level of active ERK. When the association between uPAR and integrin or integrin and fibronectin are interrupted either by reduction of surface uPAR expression, or by other means, human carcinoma cells enter a state of protracted dormancy. We show that very high levels of active ERK are required to keep cancer cells proliferating in vivo. PMID- 10684158 TI - Suppression of tumor cell growth by type IV collagen and a peptide from the NC1 domain of the alpha 3(IV) chain. PMID- 10684159 TI - [Metabolic changes in 2612 patients with nephrolithiasis]. AB - Nephrolithiasis is one of the most frequent pathologies of the urinary tract. Its prevalence in the city of Buenos Aires is 4%. Different biochemical and physiological disturbances may create an environment conductive to renal stone formation. We present the results of an ambulatory evaluation in 2612 patients for the purpose of updating the classification of nephrolithiasis. An abnormal urinary biochemistry was observed in 2423 patients (92.8%) that could be classified in 15 categories. A single diagnosis was documented in 61.5% of the patients, and the remaining 31.2% had more than one diagnosis (concurrent abnormalities). No abnormality was found in 189 stone formers (7.2%). Idiopathic hypercalciuria was the most frequent abnormality, it was encountered in 31.2%; hyperuricosuria and gouty diathesis (presence of urine pH < 5.5, with normal or high uricemia) accounted for 9.4% and 5.4% of patients, respectively. On the other hand, hypomagnesuria affected 6.7% of the stone formers and hypocitraturia was observed in 4.5%. Primary hyperparathiriodism, hyperoxaluria and cystinuria were seen less frequently in 2.6%, 1.3 and 0.45% of patients. Low urine volume was found in 12% of the patients. Among those patients with more than one abnormality, we found that hypercalciuria together with hyperuricosuria and hypocitraturia (12%) was the prevalent association followed by hypercalciuria with hyperuricosuria (9.1%). Our results show the importance of studying nephrolithiasis patients from a biochemical point of view, since this is the only way to achieve a diagnosis of the metabolic abnormality and introduce a specific therapy to prevent recurrence. PMID- 10684160 TI - [Atrioventricular node catheter ablation with condenser discharge and radiofrequency]. AB - The AV junction ablation was useful to treat patients with drug-refractory supraventricular arrhythmias. The purpose of this study was to determine short and long-term success and complications of the atrioventricular nodal catheter ablation and to compare direct current and radiofrequency energy. Forty patients underwent AV nodal ablation with direct current energy (Group I) and forty patients with radiofrequency (Group II). They were followed up for a mean of 76 +/- 49 and 28 +/- 20 months, respectively. Persistent complete AV block was successfully induced during the first ablation session in 45% of 40 patients who underwent DC energy, while in 50% it was modulated. All patients in the radiofrequency group had complete AV block. The rate of recurrence of AV conduction was 7.5% and 2.5% respectively. Immediate complications did not occur after either procedure. One patient died suddenly in each group during follow-up. AV nodal ablation with radiofrequency energy appears to be as efficacious and safe as direct current energy. PMID- 10684161 TI - [Effect of iron supplementation and its frequency during pregnancy]. AB - The iron (Fe) nutritional status of 203 healthy pregnant women was assessed at the first prenatal visit (To) (gestational age: 16.9 weeks +/- 3.81. Women were randomly assigned to one of three groups: G1 and G2 were supplemented with ferrous fumarate (60 mg elemental Fe) daily or intermittently (three times a week), respectively; and GC was the control group, without supplementation. The follow up was carried out until 34-37 weeks of gestational age (Tf), but only 43% of pregnant women completed the trial. At To and Tf fasting blood samples were collected and Hematocrit (Hct), Hemoglobin (Hb), Erythrocyte Protoporphyrin (EP) and Serum Ferritin (FERR) were determined. The percentage of women with abnormal biochemical values at To (n = 203) was: Hb (g/dl) < 10.5: 2.6%; PE (microgram/dl of red blood cells) > 70: 4.8%; FERR (ng/ml) < 10: 4.4%. Results (X +/- DE) of women that completed the follow up were at To and Tf, respectively: Hct (%): GC: 37.7 +/- 3.4 and 36.0 +/- 3.2 (p < 0.05); G1: 38.8 +/- 2.2 and 38.0 +/- 2.6; G2: 39.0 +/- 2.7 and 37.7 +/- 3.7; Hb (g/dl): GC: 12.5 +/- 1.2 and 11.9 +/- 1.3 (p < 0.05); G1: 12.6 +/- 1.1 and 12.8 +/- 1.1; G2: 12.9 +/- 0.9 and 12.2 +/- 1.5; PE (microgram/dl red blood cells): GC: 30 +/- 17 and 43 +/- 22 (p < 0.01); G1: 26 +/ 13 and 38 +/- 21 (p < 0.01); G2: 26 +/- 16 and 31 +/- 26; FERR (ng/ml): GC: 75 +/- 67 and 31 +/- 49 (p < 0.01); G1: 46 +/- 34 and 19 +/- 10 (p < 0.01); G2: 43 +/- 11 and 11 +/- 7 (p < 0.01). These results show: a) Fe administration was efficient to mitigate Hb decrease; b) Fe stores decreased during pregnancy regardless of Fe supplementation and frequency; c) EP values indicate that intermittent Fe administration was more efficient to maintain normal erythropoiesis. PMID- 10684162 TI - [Syphilis in adolescent mothers in the city of Posadas, Province of Misiones]. AB - During three months (April to June 1997) 1,238 consecutive pregnant women were studied at the time of delivery at the Madariaga public Hospital. Syphilis was confirmed in 26 (2.1%) women, and 15 cases (57.7%) of congenital syphilis were demonstrated in newborns one of whom was a stillborn. Of the syphilitic women 61.5% were 20 years old (average), 65.4% were single, 19.2% had a stable partner and 15.4% were married; 70% of them had finished elementary school (seven years), but despite this discrete level of instruction and that they were benefited with free health attention, 73% of them had not started or completed the pregnancy control. None of these women acted as sexual workers or were drug users; 57.7% were unemployed and the remainder worked as domestic servants or were still going to school. Menarca started at 13 (average) and the age of the first sexual activity was 15 (average). The distribution of the cases of syphilis within the city area shows four clusters that coincide with the lower income population, but not with marginal groups. The failure to submit to medical control during pregnancy among syphilitic women is directly linked with an increased risk for congenital syphilis. The specific prevalence of syphilis in women (20 years old or less) pregnant or not, shows an alarming hidden epidemic situation. An interinstitutional and communitary program, with direct interventions within the detected population clusters, is now underway in order to control syphilis. Undesired pregnancy and syphilis seem to be associated with adolescent unsafe sex conducts. A coordinated program between Public Health Service and National Misiones University is operating, visiting home by home, in order to decrease or eliminate congenital syphilis and is considered a priority health problem. Unfortunately, if sexual conducts do not undergo changes in the near future, at least by the correct use of condoms, HIV will replace syphilis. PMID- 10684163 TI - Hematologic study of newborn umbilical cord blood. AB - Hematological parameters in newborn umbilical cord blood samples (n = 476), collected at the Hospital Provincial del Centenario, Rosario, were studied. They were divided into 3 groups: (I) full term newborns with weight according to gestational age; (II) low weight and normal gestational age; (III) preterm newborns. The results were as follows: group (I) Hb: 15.5 +/- 1.1 g/dl; RBC; 4.66 +/- 0.33 x 10(12)/l; PCV: 49% +/- 4.3%, MCV 105.1 +/- 5.3 fl; MHC: 33.2 +/- 1.2 pg. Decreased Hb concentration (p < 0.05) and increased MCV (p < 0.01) were observed in preterm newborns in comparison with normal ones, and a slight PCV increase and RBC values in low weight newborns compared to the control group (p < 0.05). Erythrocyte morphology was normal as well as reticulocyte values in these samples. The electrophoretic pattern was (FA) with the following Hb F values 66.3 +/- 6.8%, and Hb A2 0.45 +/- 0.3% in group (I), with a significant increase of Hb F in 30-35 weeks preterm newborns. Group (I) values are considered as normal hematological parameters in newborns in our country, whereas MCV < 94.7 fl is considered as a neonatal microcytosis marker, consequently an alert to investigate alpha-thalassemia. There was no influence on Hb concentration due to maternal smoking habit. The present work could be of relevance for our region since up to the present time there are no similar records. PMID- 10684164 TI - [Vitamin D status in women living in Buenos Aires]. AB - Bone loss has both age-related and menopausal components. The causes of age related bone loss are multifactorial. In order to establish vitamin D status in women in our city (34 degrees S), calcidiol levels were assessed in 357 ambulatory women aged 40-90 years. One hundred and eighty were evaluated during summer time and 177 during winter time. We also evaluated intact PTH values in a subgroup of 231 women and this allowed us to document the prevalence of secondary hyperparathyroidism. Summer levels of calcidiol were significantly higher than in winter: 25.3 +/- 8.5 vs 21.3 +/- 7.4 ng/ml (p < 0.001). We found 4.4% of calcidiol levels < 10 ng/ml (2.2% in summer and 6.6% in winter). Prevalence of calcidiol between 10-20 ng/ml reached 67% in winter and went down to 25% during summer. Prevalence of secondary hyperparathyroidism was 5.2%. Even though prevalence of vitamin D deficiency was low, a great proportion of ambulatory women had calcidiol levels between 10-20 ng/ml. These values would not be sufficient for elderly people and could result in increased calcium mobilisation and further bone loss. PMID- 10684165 TI - [Utility of gas chromatography for the identification of mycobacterial species]. AB - The human immunodeficiency virus (HIV) epidemic has altered the epidemiological profile of tuberculosis in both industrialized and developing countries. Serious diseases caused by mycobacteria other than Mycobacterium tuberculosis, mostly belonging to the M. avium-intracellulare complex (MAC), have become very common in association with severe immunosuppression. The increase in mycobacterial disease complexity has stimulated the development of more rapid and efficient methods for diagnosis. In the present study, we investigated and assessed the suitability of a gas-liquid chromatography technique for diagnosis of clinically important mycobacteria in Argentina. An identification scheme was developed from the results obtained in a previous study where we characterized the cellular fatty acids and the mycolic acid cleavage products from most frequent species in Argentina. Of 183 isolates tested, 69% were correctly identified to species level and 5% were incorrectly classified. If we only take into account the isolates that could be identified, 93% were correctly identified. Although all of the isolates of M. tuberculosis were correctly identified, four isolates of MAC incorrectly matched by M. tuberculosis. Gas chromatography provides a rapid technique of highly predictive value for mycobacteria identification; it could be used in reference laboratories as a rapid presumptive identification until the biochemical tests are completed. PMID- 10684166 TI - Absence of mutations in the p53 tumor suppressor gene in non-invasive Cushing adenomas. AB - A lot of evidence supports the existence of a monoclonal origin for pituitary tumors, and several genetic alterations have already been confirmed as necessary or sufficient for unrestrained cellular growth and pituitary function. The p53 gene, a known tumor-suppressor gene (TSG), encodes a protein that exerts antiproliferative effects such as cell-growth arrest and apoptosis in response to several types of stimuli. In fact, several human cancers are believed to be caused by p53 mutations. In the case of pituitary tumors, p53 protein accumulation has been described in ACTH-secreting pituitary adenomas. Since increased amounts of the p53 protein are often related to mutations of its gene, we decided to explore the existence of p53 mutations in the tumor tissues of 9 patients bearing non-invasive corticotropinomas, excised by the transphenoidal route. We screened mutations in exons 5 to 8 of the p53 gene by the PCR-SSCP analysis. We were not able to find any mutation in the exons investigated. Our results are in close accordance with those obtained previously for other types of pituitary tumors. PMID- 10684167 TI - [Role of nitric oxide in the synthesis of prostaglandin F2 alpha and progesterone during luteolysis in the rat]. AB - In the corpus luteum (CL) prostaglandin F2 alpha (PGF2 alpha) is a luteolytic agent. Nitric oxide (NO) is a messenger molecule capable of modulating diverse pathophysiological processes. Many of these functions are related with the female reproductive tract. The aim of the present study was to investigate the role of ovarian NO in PGF2 alpha production arid in progesterone synthesis during CL regression in the rat. By means of the intrabursa (i.b.) ovarian sac treatment of two competitive NO inhibitors, NG-monomethyl-L-arginine (L-NMMA; 1 mg/kg); NW Nitro-L-arginine methyl ester (L-NAME, 3 mg/kg) and sodium nitroprusside (SNP, 0.05 mg/kg) as a NO generator we found that NO, produced by the ovarian tissue during the last days (days 8 and 9) of CL development, acted by increasing PGF2 alpha production in the ovary and diminishing seric progesterone levels leading to CL involution. We also postulated a positive feedback mechanism between PGF2 alpha and NO, to ensure luteal regression. Thus, we injected intraperitoneally (i.p.) a luteolytic dose (3 micrograms/kg) of a synthetic PGF2 alpha during the mid and late phase of CL development. The ovarian activity was evaluated. The results confirmed our hypothesis; we did not see any effect in mid-stage of CL development, while at a late stage enhancement of ovarian NOs activity was observed in PGF2 alpha-infected animals. PMID- 10684168 TI - [Anti glomerular basement membrane disease in a renal transplant patient with Alport syndrome]. AB - We report a case of anti GBM disease that developed in the renal graft of a patient with Alport syndrome. After reaching abnormal values of creatinine, the patient presented with deteriorating renal function three months after a cadaver transplant and the biopsy showed crescent formation, and linear IF deposits. Circulating antibodies against alpha 5 chain of type IV collagen were found and plasmaphereses stabilized the condition for one year until a lung infection led to withdrawal of the immunosuppressive drugs and the patient returned to dialysis. We discuss the possible mechanisms underlying the specificity of the circulating antibodies in this case, which differs from the target characteristic of the idiopathic form of anti GBM disease, the alpha 3 (IV) chain. PMID- 10684169 TI - [Rosacea, vinegar and lemon, dysuria and Helicobacter pylori]. AB - A 52 year old male consulted his clinician because of dysuria, difficulty in voiding and cutaneous lesions that were cured with high daily ingestion of acid substances (vinegar and lemon). For the last 20 years he had made several consultations without finding any solution to his problem. The patient was advised to stop acid ingestion after which he presented disuria and skin lesions compatible with rosacea. Due to the known association between this skin disorder and gastric colonization with H. pylori an upper gastrointestinal endoscopy was performed and the presence of the bacteria was confirmed. The patient received specific treatment with permanent resolution of the symptoms. We repeated the endoscopy with biopsy that did not show the presence of H. pylori. It is suggested that gastric colonization with H. pylori could be related to irritative symptoms of the lower urinary tract that are not due to other disease processes. PMID- 10684170 TI - [Bronchiectasis, endothoracic goiter and hemoptysis]. PMID- 10684171 TI - [Biology of heat shock proteins]. AB - Hsp (Heat shock proteins) are a family of constitutive proteins of all pro and eukariotic cells that play different physiological roles: they promote the folding (acquisition of tertiary structure) assembly, translocation and secretion of newly synthesized polypeptides and participate in the removal or repairing of denatured proteins acting as molecular chaperons. This family of proteins is composed by numerous members grouped according to their molecular weight. When cells are subjected to different stresses such as hyperthermic shock, radiation, toxins, viral infections, etc., Hsp are overexpressed. In this way, they exert a cytoprotective effect, making the cells resistant to apoptosis. In humans, Hsp are overexpressed in cancer cells from ovary, endometrium, breast, prostate, digestive tract, etc. In some cases, overexpression is correlated with an unfavorable outcome because these proteins could favour metastatic disease. Some authors associate them not only with proliferation but also with differentiation of the neoplastic tissue. Recent studies show their influence in resistance to chemotherapeutic drugs. In autoimmune diseases like rheumatoid arthritis, Hsp can suppress the inflammatory response. Nevertheless, their role in the immune system has not been well established. PMID- 10684172 TI - [Family violence. The physician's role]. AB - Family violence is mostly used against women. It is the cause of heavy psychological and physical damages as well as social and economic costs. It also has a repercussion on the future generations. Medical practitioners should be prepared to recognise these situations in order to give support, counsel and assistance to victims of abuse. Training in family violence should be included in pre and post courses in Medical School programs. PMID- 10684173 TI - [Participation of transcriptional factors in the regulation of lymphocyte differentiation]. AB - The cytomorphological changes as well as the expression of certain markers in the different stages of lymphocyte differentiation are well known. Studies carried out on expression patterns of specific genes in lymphoid cells and their regulatory mechanism have led to the identification of the fundamental mechanism of cell development and activation. A great deal of knowledge has accumulated concerning enhancers and promoters, the regulatory elements of those genes and the transcriptional factors to which they bind. The present paper analyzes these components and the participation of some of them such as PU.1, Ikaros, Aiolos, GATA-3, Egf-1, E2A, EBF-1, PAX-5 (BSAP), TFE-3, Oct-1, Oct-2, and Nf-kappa B in the regulation of the differentiation stages of cells belonging to the B and T lymphoid lineages. PMID- 10684174 TI - [Rethinking the pathogenesis of chronic chagasic cardiopathy at the closing of the millennium]. PMID- 10684175 TI - [Recommendations on the ethics of withholding and/or withdrawing life-sustaining treatment in the critically ill. Comite de Bioetica de la Sociedad Argentina de Terapia Intensiva]. PMID- 10684176 TI - [Addio del passato. What kind of memory is musical memory?]. PMID- 10684177 TI - [Procedures in abattoirs and meat inspection. State of the discussion on the implementation of alternative systems]. AB - Results of field studies which were performed in EU member states and which dealt with meat inspection procedures in fattening pigs were reviewed with respect to their contribution to alternative systems of meat inspection. The following was concluded: Inspection ante mortem (monitoring of circumstances at the farm of origin): Data which should be given to the notice of the authorities have not yet been satisfactorily defined and their informative value still is not yet clear The predictive character of information from the farm of origin regarding the health status of the animals (results of meat inspection) is not yet sound enough The technique of ante mortem inspection should be reconsidered also. Meat inspection (monitoring and assessment of lesions): In all studies the compared techniques (visual and official procedure) did not entirely find all lesions, which were supposed to be on the carcasses and organs. This was true in different percentages. The impact on consumer's health from the lesions monitored has to be discussed more thoroughly. The information presently available is not yet sound enough to generate a new practicable system of meat inspection. Further pilot studies are necessary. PMID- 10684178 TI - Effect of avilamycin, tylosin and ionophore anticoccidials on Clostridium perfringens enterotoxaemia in chickens. AB - In order to study the prophylactic and metaphylactic effect of antomicrobial growth promoters and ionophorous anticoccidials on the incidence of Cl. perfringens enterotoxaemia in chickens, experimental attempts were performed with 675 chickens in 27 trials. The birds were intraduodenally infected with Cl. perfringens type A (ATCC 3624). The following antimicrobial growth promoters and ionophore anticoccidials were used either on their own or in combination: avilamycin, narasin, monensin and tylosin. While infected and non-medicated trials showed an average incubation period of 1 week, clinical symptoms occurred 2-4 days later in infected and medicated birds. Avilamycin medicated birds had the longest incubation period. In the infected and non-medicated trials, a mortality rate of 16%-36% was noted within 3 weeks post infection. The avilamycin trials showed a mortality rate of 0-8% (0-2 birds died) and the narasin and monensin a mortality rate of 0-8%, respectively. In the combination groups (monensin + avilamycin or narasin + avilamycin), the mortality rate ranged from 0 to 4%. Tylosin showed a very good metaphylactic/therapeutic effect against Cl. perfringens enterotoxaemia. Following infection, medicated birds showed a significantly better bodyweight gain than the chickens, whose feeds had not been supplemented. From epidemiological point of view, the systematic prevention of coccidiosis is a key in the control of Cl. perfringens enterotoxaemia in chickens. PMID- 10684179 TI - [Effect of mycotoxin contaminated feed on production parameters of dairy cows]. AB - Based on a feeding trial using 27 lactating "Simmental-cows" the effect of naturally contaminated feed with deoxynivalenol (DON) as well as zearalenone (ZON) regarding production parameters was examined. 3 groups of cows according to lactation number, milk yield (kg ECM) and body mass were used. The average daily intake of DON in group K was 12.4 mg, in group T 14.1 mg and in group M 14.3 mg and ZON in group K was 12.4 mg, in group T 0.67 mg and in group M 0.68 mg respectively. The feed of animals of group M was supplemented with "Mycofix Plus" as mycotoxin inactivator. The red and white blood picture including the thrombocytes were in all groups within the normal range. Concerning enzymes (GGT, AP) and metabolites (GLUC, TBIL, UREA, CREA) the mean values of the 3 groups were in the normal range. Slightly increased were the mean values of all groups in respect to the AST- and GLDH-activities. Volatile fatty acids of the rumen content were significantly highest in group M, also the number of dead rumen infusoria was significantly decreased, but the counts of small sized infusoria increased. The study has shown that "Mycofix Plus" might be able to enhance the activity of rumen flora concerning detoxification of mycotoxins in feed of dairy cows. PMID- 10684180 TI - [Experience with simple ELISA test systems for Brucella serology in cattle, sheep and goats]. AB - The objective of this work was to use the ELISA technique for the serological surveillance for freedom of brucellosis of cattle, sheep and goats. By comparing 28 cattle sera taken after a brucellosis outbreak, 15 bovine sera supplied by the Federal Institute for Health Protection of Consumers and Veterinary Medicine (BgVV) and 497 serum slow agglutination test (SSAT) and complement fixation test (CFT) negative bovine sera from herds officially declared free of brucellosis, the ELISA technique not only shows higher sensitivity as compared to SSAT and CFT but also distinguishes clearly between positive and negative reactions. The serological comparison by SSAT, CFT and ELISA of 615 cattle, 624 sheep and 630 goat sera from herds acknowledged as brucellosis free showed equivalent specificities for both CFT and ELISA. The specificity of the SSAT was much lower, 81.1% in cattle and 96.2% in goat sera. The examination of 5796 cattle, 1337 calf, 5031 sheep and 1796 goat sera demonstrates the advantage of the ELISA technique as routine method. The possible application of the ELISA technique as a screening method for serological brucellosis tests in sheep, goats and possibly also in pigs is discussed. PMID- 10684182 TI - On the presence of ganglion cells in the intracranial portion of the accessory nerve (XI cranial nerve) in some mammals. AB - The intracranial tract of the accessory nerve (XI cranial nerve) was studied in some mammals (equines, domestic and wild ruminants, pig, carnivores, rabbit, nutria, guinea pig, hamster, hedgehog). The specimens were embedded in paraffin or paraplast, the sections were stained with cresyl violet, haematoxylin and eosin, or submitted to argentic impregnation. Pseudounipolar ganglion cells were found in all the mammals examined, with the exception of the cat. The number of cells and their variability in the different species and subjects were related. The topography and morphology of the cells were described. This comparative study has demonstrated that the accessory nerve is not a entirely motor nerve, but it is a mixed, motor and sensitive, nerve. Nevertheless, we think further studies are necessary in order to establish the peripheral distribution, the central pathway and the functional role of the pseudounipolar neurons found in the intracranial tract of the accessory nerve. PMID- 10684181 TI - Intertwined Sharpey fibers in human acellular cementum. AB - We had carried out a detailed morphological study on the human acellular extrinsic fiber cementum (AEFC) in order to support the exclusively extrinsic origin of matrix collagen bundles. Mesial and distal cervical third of fresh premolars from young individuals were examined. Semi-thin and thin section clearly show the Sharpey fibres entering in the cementum at right-angle to the root surface and coursing throughout the cementum to the cemento-dentinal junction. On their way to the dentin the Sharpey fibres divide into smaller bundles which, coursing obliquely or tangentially, intersect with others deriving from neighbouring Sharpey fibres. Both de-proteinated and decalcified samples observed at SEM present Sharpey fibres along the fractured surfaces entering and running trough the cementum perpendicularly to the root surface. Fibril bundles are seen branching out from the main body of a single Sharpey fibre and coursing obliquely or perpendicularly to the original fibre. These morphological evidences obtained both at TEM and SEM further confirm that in AEFC fibril bundles running parallel or obliquely to the root surface are branches of Sharpey fibres and not intrinsic cementum fibres. PMID- 10684183 TI - Bone mineral density and anthropometric measures in normal and osteoporotic men. AB - In an ethnically homogeneous men population living in Tuscany, Italy, the relationship between age, height, body weight and bone mineral density were studied. In 50 men bone mineral density was measured by Dual Energy X ray Absorptiometry (DEXA). 13 subjects (26%) were osteoporotic. Age and bone mineral density were not related (R2 = 0.052). Bone mineral density was associated with body weight (R2 = 0.303), and height (R2 = 0.155). In osteoporotic men, mean (+/- SD) body weight was Kg. 65.8 +/- 11.2, lower than that recorded in non osteoporotic men, Kg. 77.3 +/- 10.2, (p = 0.0013). Age in osteoporotic and non osteoporotic men did not differ (53.8 +/- 13.6 yrs and 60.9 +/- 11.8 yrs, respectively; p = 0.077). In conclusion, anthropometric factors, as predictors of bone disease, behave differently in women and men. PMID- 10684184 TI - Placental morphometry in pregnancies complicated by intrauterine growth retardation with absent or reversed end diastolic flow in the umbilical artery. AB - The aim of this study was to assess any possible correlation between villous tree architecture and its vascularization, and absent or reversed end-diastolic flow velocity (ARED) in the umbilical artery. The study group included seven pregnancies complicated by IUGR (estimated fetal weight < 10th percentile) and absent end-diastolic flow velocity in the umbilical artery. A gestational-age matched group of seven normally grown pregnancies was selected as control group. At delivery, the placenta was weighed and immersed in 10% neutral buffered formalin. A stratified random sampling procedure was used to obtain 12 blocks of full-thickness tissue per organ. A single random section was cut from each block. The following morphometric parameters were evaluated in each section: mean vessel diameter, volume density of the villous tissue, stem villi and terminal villi. Measurements were performed using a computerized Video Image Analysis system. No significant difference in mean vessel diameter was found between the two groups (37.1 microns versus 36.1 microns; p = 0.1). There was a significant reduction in the proportion of total villous tissue in the ARED group (43% versus 52%): this was due to a significant reduction in the volume of tissue occupied by the terminal villi (14.1% versus 18.4%). No significant difference was found in the proportion of villous tissue occupied by the stem villi (42% versus 40%). Several studies have investigated the anatomical and/or vasomotor bases of absent end diastolic flow velocity in the umbilical artery of fetuses with severe IUGR. Our observations of a significant reduction in the proportion of villous tissue occupied by the peripheral villi are consistent with the theory that failure of normal development of the terminal villous is responsible for the increased vascular resistance in IUGR pregnancies with ARED. PMID- 10684185 TI - [Treatment of acute destructive alimentary pancreatitis]. AB - The paper reports treatment of 52 patients with acute destructive pancreatitis for the period from 1993 to 1999. Up to 1997 conservative policy for management of such patients was used, operative treatment was carried on only in pyoseptic complications (43 patients, group 1). Later on early video-endoscopic procedures (9 patients) aimed at removal of toxic exudate from the abdominal cavity, its sanation and drainage with the use of not less than 5 drainage tubes were employed. From 43 patients of group 1 10 died. There were no lethal outcomes in group 2. The authors believe that early evacuation of the toxic purulent exudate with lavage and drainage of the abdominal cavity promotes more effective elimination of multiorganic insufficiency and decreases the risk for development of pyoseptic complications. PMID- 10684186 TI - [Humoral immunity and antigens of HLA system in acute pancreatitis]. AB - In the authors investigations patients with acute pancreatitis had antibodies to endogenic antigens: to o-DNA--in 58.5%, to d-DNA--in 53.7%, to n-DNA in 51.2%, to trypsin--in 42.7%, to insulin--in 28.1% and to the tissue antigen of the pancreas -in 19.5%. A rise of serum immunoglobulins' and circulating immune complexes' levels were established. Alterations of humoral immunity in connection with high frequency of antigens HLA A1, B8, B18 associated with disregulation among T- and B-links of immune system detected in patients with acute pancreatitis, represent genetic and humoral mechanisms which mediate the development of autoimmune reactions in this disease. PMID- 10684187 TI - [Management of large and giant postoperative ventral hernias]. AB - The experience of the Department for Plastic Surgery of the A.V. Vishnevsky. Institute of Surgery of RAMS in surgical treatment of 262 patients with postoperative hernias of the anterior abdominal wall has been summarized for the period from 1993 to 1998. 138 patients (test group) had large and giant hernias, 124 (control group)--small and middle-sized postoperative hernias. In large and giant hernias more frequently (46.1%) combined plastic reconstruction of the anterior abdominal wall was used rather than in small and middle-sized hernias (21.5%). In the majority of patients in control group (78.5%) autoplasty of the abdominal wall defect was carried out. In the long term period relapses developed in 9 (7.0%) patients of the test group. In 7 cases the relapse developed after the autoplasty (10.1%), and in 2 cases--after the use of combined method (3.4%). This supports perspectiveness of the method of postoperative hernioplasty by the combined method. Lethality rate after hernioplasty in the test group made up 2.3% (3 patients). PMID- 10684188 TI - [Surgical treatment of paracolostomu hernias and prolapses]. AB - The results of surgical treatment for paracolostomy hernias and prolapses in 71 patients show insufficient effectiveness of this method for surgical correction of such paracolostomy complications as paracolostomy hernias and prolapses: the rate of their relapses reaches 50%. The authors have developed a new effective method for surgical treatment of complicated colostomy: intraabdominal reconstruction of colostomy with retroperitoneal pull and laying of prestomal segment of the sygmoid colon. PMID- 10684189 TI - [Lychtenstein's hernioplasty in inguinal hernmias]. AB - The choice of plastic treatment of inguinal hernias is stile a problem. The rate of relapses in conventional methods of hernioplasty averages 8%. The commonest complications are nerves' injuries and damage to the arterial and venous vessels of the spermatic cord. The hope for the decrease of complications rate is based on implementation of "unstretched" surgery. The rate of relapses after endoscopical hernioplasties varies from 0.8% after transabdominal preperitoneal hernioplasty to 3.2%--after intraperitoneal one. Laparoscopic methods for hernioplasty are technically complicated and expensive. The authors used Lychtenstein's hernioplasty as an alternative to laparoscopic hernioplasty methods. The technique is described according to which 76 operations were carried out. The method is much easier than laparoscopical one. The course of the early postoperative period is comparable with that one after laparoscopical operations. It is stressed that Lychtenstein's hernioplasty should be considered as a method of choice in majority of patients with inguinal hernias. PMID- 10684190 TI - [Ultrasound examination in patients with peritonitis]. AB - Ultrasonographic features of developed intraabdominal complications were in 73 patients, in 43 of them being local and in 30-diffuse peritonitis. The diagnosis was based on the complex manifestations of' sonographic changes of the small bowel and other organs of the abdominal cavity which the authors suggest to interpret as an ultrasound syndrome of "the bowel deficiency". The possibility of dynamic ultrasound control in postoperative period for registration of the features indicating elimination or progressing of peritonitis is shown. PMID- 10684191 TI - [Provisional ileostomy as a treatment of postoperative purulent peritonitis]. AB - Early elimination of functional bowel obstruction in postoperative diffuse purulent peritonitis (PDPP) is an important factor determining the outcomes of the disease. Nasogastrointestinal drainage used for this purpose is capable to adequately aspirate small bowel contents from the segments located at a distance not farther than 80-100 cm. In the majority of patients with PDPP, lacking of vital capacity of bowel wall as well as insufficiency of previously laid sutures were revealed, which forced a surgeon to resort to resection; in such cases the method of choice for decompression should be the application of terminal ileostomy. Ileostomy in patients with PDPP provides adequate decompression of the bowel. The optimal terms for elimination of ileostomy is 3-4 weeks postoperatively. PMID- 10684192 TI - [Ultrasound examination for diagnosis of postoperative peritonitis]. AB - 78 patients with peritonitis after various operations on the organs of abdominal cavity were followed up. Ultrasound examinations (n = 86) were carried out in 59 patients. In 28 of them US has revealed abscesses of the abdominal cavity. Comparative analysis showed coincidence of the diagnosis of peritonitis by clinical and roentgenological data with the results of relaparotomy in 10 (58.8%) of 17 examined patients. The same coincidence of the diagnosis of peritonitis by clinical, roentgenological and sonographic data on one side and of the results of relaparotomy on the other was found in 48 (81.4%) of 59 patients with peritonitis. It is stated, that US examination considerably increases the effectiveness of the diagnosis of postoperative peritonitis and incapsulated cavities. PMID- 10684193 TI - [Management of stomach cardial ulcers complicated by bleeding]. AB - The results of treatment of the patients with bleeding ulcers of cardial part of the stomach have been analysed. In contrast to gastric ulcers of another location, cardial ones are poorly respond conservative treatment, are usually accompanied by severe complications, and are more frequently prone to malignant transformation. New method for proximal resection of the stomach has been developed, which was successfully used in 14 patients. There were no complications and lethal outcomes. Yearly and long-term results were favourable. PMID- 10684194 TI - [Gastrectomy with creation of small intestinal reservoir]. AB - A new variety of the esophago-intestinal anastomosis in gastrectomy is presented. The application of the anastomosis begins with formation of the cuff around the esophagus by the usage of the afferent and efferent loops, which then being joined together in a shape f reservoir. A total of 113 patients with cancer of the stomach (83), cancer of the gastric stump (10), giant ulcers of the cardial part of the stomach complicated by bleeding (15), and Zollinger-Ellison (5) syndrome have been operated on. Insufficiency of the anastomosis was revealed in 2 patients (1.8%). 6 patients (5.1%) died, 1 patient with insufficiency of the esophageal anastomosis included. There were no clinical manifestations and objective features of reflux-esophagitis after the usage of the suggested method for anastomosis, the reservoir function after the resection of the stomach was for the most part compensated. PMID- 10684195 TI - [Therapeutic policy in rectal cancer]. AB - The results of clinical examination and treatment of 751 patients with rectal cancer are presented. A total of 684 (91.0%) patients underwent radical surgery. In 439 (64.2%) from them sphincter saving operations were carried out. The rate of sphincter saving procedures for the last 5 years has increased up to 73.8%, most often of them transbdominal resection was used--284 (64.7%) patients. Sygmoidorectal anastomosis in 158 (55.6%) patients was carried out by the hand sutures, and in 126 (44.4%)--suturing apparatuses were used. The operation was finished by the suture of the pelvic peritoneum by two layers of sutures and the drainage of the area of the anastomosis from the perineum site by two tubes for prevention of the development of peritonitis even in case of insufficiency of the anastomosis was made. All the operated patients underwent preoperative radiotherapy with a total dose 20-25 Gy. The basic stage of the operation was on demand followed by chemohyperthermic perfusion of the abdominal cavity. It was carried out in 30 patients, including 12 patients with rectal cancer. In cancer of the rectum of the II B and III stages the usage of chemotherapy is obligatory. New operations for metastases of cancer of the colon into the liver were made in 4 patients (left and extended right hemihepatectomy). Complications developed in 33.5% of patients, lethality made up 3.8%, including 1.8% for the last 5 years. PMID- 10684196 TI - [Advisability of combined operations in proctologic patients]. AB - The analysis of surgical treatment for combined diseases of the anorectal region in 106 patients is presented. The mean duration of the hospital stay after one stage procedures made up 10.8 days, whereas in proctological patients with a single disease it made up 9 days. The individual approach to the choice of surgical policy and keeping to the order of the stages of the operation are essential in treatment of patients with combined diseases. The results of the study support the use of combined operations in patients with disease of the colon and the anorectal area. PMID- 10684197 TI - [Radiofrequency electrostimulation of sympathetic nerve boundary trunk in patients with bronchial asthma]. AB - In 7 patients aged from 43 to 64 years with severe infectious allergic forms of bronchial asthma with curative aims radiofrequency electrostimulator for the boundary trunk of the sympathic nerve (BTSN) has been implanted in the neck. Daily sessions of electrostimulation (ES) by the current with parameters 1-100 Hz, 0.1-0.4 m/s, 0.1-2.0 V adjusted individually by maximal broncholytic and clinical effects were performed for 5 years. It was established that ES of BTSN can produce both bronchial dilatation and bronchial spasm. Application of ES of BTSN has resulted in a decrease of bronchial hyperreactivity, frequency of asthmatic attacks 2.6-3.2 fold in consumption of antiasthmatic drugs 2.4-2.7 fold. The only complication has been detected--rejection of the electrostimulator due to the defect of its capsule. This method can be applied as an adjuvant in therapy of severe forms of bronchial asthma resistant to drug treatment. PMID- 10684198 TI - [Systemic inflammatory response syndrome and sepsis in maxillofacial surgery clinic]. AB - A total of 106 patients with surgical infection in maxillofacial region were studied. Clinical and laboratory diagnostic data on surgical infection and sepsis in maxillofacial surgery are presented. Integral indices of blood circulation, respiration, brain and liver metabolism, regarding severity of surgical infection were determined. Pathogenetically substantiated therapy of sepsis is proposed. PMID- 10684199 TI - [Registration of superficial acoustic waves velocity (SAWV) in prognosis and diagnosis of wound healing]. AB - A new method of objective diagnostic express-monitoring of a wound healing course has been developed for prognosis and evaluation of effectiveness of the treatment for various purulent wounds. The method is based on measurement of SAWV in the skin integument surrounding festered wound. Portable device-acoustic skin analyser (ASA) was used for the measurement of SAWW. The following results were obtained in 273 patients: in phase I of healing process in the first period SAWV was 35.0-49.9 m/s; in the second period of phase I of wound healing it was 50.0 90.0 m/s; in phase II--35.0-49.0 m/s; in phase III--25.0-34.9 m/s. SAWV in healthy skin is 18-24.9 m/sec. These measurements of SAWV completely correlated with cytological and hystological examination data. PMID- 10684200 TI - [Up-to=date view on medical documentation in surgical clinic]. AB - Computer variants of the fragments of clinical records were developed. They represent the parts: "Title-page", "Examination of surgical patient in the admission department", "Protocol of laparoscopic cholecystectomy". During modelling of the intellectual contents of the modules the principle of the formalized protocol was used, which has been realized with use of a context depending menu. According to the authors opinion, newly developed programs provide objective and correct reflection of any clinical and surgical situation, use of standardized terminology and classifications, save the surgeons the trouble of "scribbling" and decrease time-consuming registration of medical records, provide specialized information, prevent possible diagnostic and technical errors, and give physicians, legal defence. PMID- 10684201 TI - [Extra-anatomical shunting of femoral artery in purulent wound]. PMID- 10684202 TI - [Cancer of appendix complicated destructive appendicitis]. PMID- 10684203 TI - [Isolated injuries of gallbladder]. PMID- 10684204 TI - [Patient's quality of life after surgical treatment]. PMID- 10684205 TI - [I.M. Popov'ian - physician, researcher, public figure]. PMID- 10684206 TI - [Angina pectoris, left ventricular dysfunction and age]. PMID- 10684207 TI - [Atherosclerosis: some topical issues of pathogenesis, diagnosis, treatment and prevention]. PMID- 10684208 TI - [Autoimmune and rheumatic aspects of HCV infection (lecture)]. PMID- 10684209 TI - [Present-day infectious endocarditis (part i)]. PMID- 10684210 TI - [Treatment of arterial hypertension in patients with bronchial asthma]. AB - Treatment of arterial hypertension in patients with concurrent chronic obstructive diseases of the lungs has limits in prescription of antihypertensive drugs. STAMLO--long-acting calcium antagonist--is proposed for monotherapy of mild and moderate hypertension in patients with bronchial asthma. The drug decreases hypertension in the greater and lesser circulation, improves bronchial permeability. PMID- 10684211 TI - [Cellular activity in healthy subjects with cardiovascular integral risk factors]. AB - The study was made of permeability of erythrocytic membrane (PEM), electrokinetic properties of cellular nucleus (ECN) and phagocytic activity of neutrophils (PAN) in whole blood of 130 healthy subjects with integral risk factors of cardiovascular disease. First-level test immunogram was analysed. Hypercholesterolemia and hyperbetalipidemia were associated with high PAN. Obese patients with cardiovascular hereditary load had elevated PAN and PEM. In smokers PEM was elevated but ECN was subnormal, PAN rose in parallel with number of cigarettes smoked for a day. Combination of the risk factors led to low activity of the nucleus and high PAN. It is concluded that healthy subjects with integral risk factors develop changes in functional cell activity. PMID- 10684213 TI - [Polymagnetolaser correction of vegetative and bioelectric imbalance in patients with duodenal ulcer]. AB - 90 patients with duodenal ulcer in exacerbation were divided into two groups. Control patients have received drugs against Helicobacter pylori (Hp). In addition to it, the test subjects were exposed to transcutaneous and reflex magnetolaser impact with consideration of baseline vegetative and bioelectric balance as shown by mathematical analysis of cardiac rhythm and Nakatani acupuncture diagnosis, monitoring of biopotentials of biologically active points. Impacts on general regulation systems in combined differential therapy of ulcer shorten ulcer healing, promote eradication of Hp, improve the condition of gastroduodenal mucosa. PMID- 10684212 TI - [Role of bronchospasm in impairment of bronchial permeability in bronchial asthma in attack-free period]. AB - Lung capacity (LC), functional lung capacity (FLC), FEV1, FEV1/LC%, CO25, CO50, CO75 were measured in 56 patients with bronchial asthma in attach-free period before and after inhalation of one-dose berotec and one-dose atrovent. Bronchospasm was registered in 66% of the patients. It was the only cause of bronchial obstruction in 12% of the patients and one of the causes in 54% of them. Contribution of the bronchospasm to development of bronchial obstruction was, as a rule, dominating. This contribution reached 81-100% in 60% of the patients. PMID- 10684214 TI - [Features of angina pectoris in patients with non-insulin-dependent diabetes mellitus (NIDDM)]. AB - Detailed clinicofunctional examination of anginal patients with NIDDM using variation pulsometry and electrophysiological cardiac tests in 110 patients (89 females and 21 males aged 36-79 years, mean 64.3 +/- 5.2 years) has revealed that atypical cardialgia, arterial hypertension, obesity was present in 28.1, 76.4 and 85.4% of anginal patients with NIDDM. Sympathicotonia was registered in autonomic heart regulation of 83% of them. Anginal patients exhibited also suppression of sinus node automatism and sinoatrial conduction. When NIDDM is severe, pacemaker activity of the sinus node intensified suggesting development of cardial diabetic neuropathy. PMID- 10684215 TI - [Vegetative regulation of arterial pressure in patients with arterial hypertension and non-insulin-dependent diabetes mellitus (NIDDM)]. AB - 72 patients with NIDDM (duration 5.3 +/- 3.1 years) aged 41-60 years were examined. They had also mild or moderate hypertension (duration 12.1 +/- 4.5). Control groups consisted of 15 NIDDM patients free of hypertension, 15 hypertensive patients without diabetes, 15 healthy subjects. All the patients have undergone 24-h monitoring of arterial pressure (APM) and computed cardiointervalography (CIG). As shown by CIG, hypertensive patients with NIDDM had sympathotonia which resulted from hypofunction of parasympathetic and hyperfunction of the sympathetic nervous systems. APM demonstrated enhanced variability of arterial pressure and its inadequate fall at night in patients of this group and normotensive patients with diabetes. PMID- 10684216 TI - [Efficacy of emoxipin in treatment of arterial hypertension in the Far North]. AB - A total of 95 hypertensive men working in duty regimen in the extreme North were examined. The men were found to have lipid metabolism disorders, activation of lipid peroxidation, decreased antioxidant defense in red cell membranes in all the examinees vs control hypertensive men living in moderate climatic zone. Effect of 2-week monotherapy with emoxipin was studied in 29 males with mild arterial hypertension free of obesity and coexisting diseases. Placebo was given to 10 patients. Emoxipin had an antihypertensive and antioxidant effects, improved structural-functional condition of erythrocytes in hypertensive patients. PMID- 10684217 TI - [Effect of ACE inhibitor (ednit) on portal hemodynamics in patients with hepatic cirrhosis]. PMID- 10684218 TI - [New approaches to prevention of stomach cancer in employees of chemical plants]. PMID- 10684219 TI - [Long-term atrovent treatment of chronic obstructive bronchitis]. PMID- 10684220 TI - [Medicine in Academy of Sciences in "century of Enlightenment" (the 275th anniversary of Russian Academy of Sciences)]. PMID- 10684221 TI - [There is something wrong in the studies of mammography! No support for the conclusions on benefits of breast cancer screening]. PMID- 10684222 TI - [Incidence and mortality of breast cancer during a 25-year period. International and regional comparisons]. PMID- 10684223 TI - [Spontaneous premature birth: physiopathology, predictors and management. The frequency is constant--early detection can improve therapeutic possibilities]. AB - Premature birth is associated with increased perinatal morbidity and mortality. Spontaneous premature birth can be understood as a syndrome with a number of underlying causes including infection, maternal stress, uterine distention, placental hypoxia, bleeding and lack of prostaglandin dehydrogenase. Infection is probably the most important factor at low gestational age, with uterine distention and maternal stress increasing in significance further on. In the future we may become better able than we are at determining the specific reason in each individual case, which may lead to the development of more effective treatment. On the other hand, results have been very poor with respect to prevention, and in some countries we even note a slight increase in incidence. Although premature birth is often impossible to prevent, early detection and tocolytic therapy can delay labor long enough to permit effective corticosteroid therapy and transport when required to the appropriate obstetric clinic for optimal neonatal care. PMID- 10684224 TI - [Fetal virus infection a risk factor of diabetes mellitus type 1 in children]. AB - The incidence of type-1 diabetes mellitus is high and increasing in Sweden. The etiology of IDDM is complex: several genes appear to interact with each other and with various nongenetic risk factors to induce and complete the autoimmune destruction of beta-cells. IDDM has its onset most often in childhood, and during the past decade incidence is increasing specifically among children under the age of five. Factors initiating the process should thus be sought in early life. A number of recent studies have identified perinatal events as risk determinants; among these, fetal viral infections may be causally related to the disease and may become targets for intervention and prevention. PMID- 10684225 TI - [Perinatal factors can be risk factors of diabetic nephropathy]. AB - This article discusses the association of perinatal risk determinants and the future development of diabetic nephropathy. A low birth-weight seems to increase the risk for future cardiovascular disease, hypertension and insulin resistance, all of which are features of diabetic nephropathy. In a nation-wide case controlled study we found that smoking during pregnancy and low maternal education, rather than low birth weight per se increase the risk of developing incipient nephropathy in offspring with type-1 diabetes. These factors are in addition to, and independent of, a familial disposition for cardiovascular disease and hypertension. Persistent hyperglycaemia is a prerequisite for the influence of these factors. Our findings support the hypothesis of a multifactorial aetiology of diabetic nephropathy. PMID- 10684226 TI - [When insight on disease is halting. I. Clinical assessment of impaired insight- current research summarized]. AB - This review explores the definition, assessment and possible restoration of impaired insight in psychosis. Hypotheses concerning the neuropsychological correlates of insight impairment are discussed. The distinction between impaired insight in psychosis and impaired awareness and psychological denial in other medical conditions is emphasised. PMID- 10684227 TI - [When disease insight is halting. II. The neurobiological underpinnings]. AB - This review examines the neurobiological underpinnings of impaired insight and awareness in psychosis. Crucial mainly dopaminergic pathways are discussed in relation to delusions and hallucinations. Thirty percent of psychotic patients with impaired insight will not respond to traditional neuroleptic drug treatment, and the possibility that multifocal brain etiology might underlie such cases is discussed. It is suggested that severe cases of thought process disorder in psychosis may result from such multifocal processes. PMID- 10684228 TI - [Don't miss prolactinoma! On the importance of measuring prolactin and how it is done]. AB - Prolactinoma is the most common functional pituitary tumor. Since appropriate treatment is often pharmacological rather than surgical, all patients with a tumor within or close to the sella turcica should be evaluated for prolactinoma before being sent for neurosurgery. Preanalytical factors affecting serum prolactin concentration should be taken into account when planning blood sampling. Diagnostic laboratories should aim for the use of common international calibrators and a common unitage for expressing concentrations. Assays should be carried out in such a way that the risk of falsely high or low values is minimized. Any patient with high values due to an immunometric method should be evaluated for the possible presence of endogenous antibodies against prolactin ("macroprolactinemia"). PMID- 10684229 TI - [Europe without borders--who will take care of the protection against infections?]. PMID- 10684230 TI - [Scientists and humanists can bridge over the differences of their specialties]. PMID- 10684231 TI - [A reflection on the effect of medical knowledge on man]. PMID- 10684232 TI - [Future of school health services. Concentrate on the good parts of Swedish health care!]. PMID- 10684233 TI - [How really dangerous is spinal manipulation?]. PMID- 10684234 TI - [Halting primary health care--what does the Medical Society do?]. PMID- 10684235 TI - [To master severe epilepsy requires multicompetent measures]. PMID- 10684236 TI - [Efficient cytologic screening: selective and in combination with HPV test]. PMID- 10684237 TI - [Automated quantitative image cytometry of bronchial washings in suspected lung cancer: comparison with cytology, histology and clinical diagnosis]. AB - INTRODUCTION: Automated image cytometry represents a new method for the quantitative analysis of nuclear structure and DNA-content of exfoliative airway epithelial cells. In the present investigation, we examined the correlation between automated cytometry, conventional cytology and histopathology with the final diagnosis as the "gold standard". METHODS: In 142 patients (100 males and 42 females) with suspected lung cancer and 50 controls (COPD, asthma), bronchial washings (5-10 ml) were obtained during bronchoscopy before taking biopsies for cytological and/or histological examinations. The washings were collected in 20 ml Saccomanno's fixative and centrifuged (500 g, 15 min). The cell pellet was resuspended in Saccomanno's solution. Two specimens were stained according to Papanicolaou and another two using the Feulgen reaction with thionine. Image cytometry was performed by means of a special, trainable classifier for exfoliative cells of the respiratory tract, using the Cyto-Sacant (Oncometrics, Vancouver). RESULTS: In the patients with suspected lung cancer we found numerous abnormal nuclei in 97 samples, 36 samples contained normal cells only, and 9 samples were insufficient. In our control group there was no sample with abnormal nuclei, and all washings were evaluable. Compared to the final diagnosis of lung cancer, we found a sensitivity of 90% (92/102) and a specificity of 84% (26/31). For histology sensitivity was 91% (73/80) and specificity 100%, while we found a sensitivity of 92% (92/100) and specificity of 100% for cytology. For automated cytometry the positive predicted value was 95%, the negative predicted value 71%. CONCLUSIONS: In the investigation of patients with suspected lung cancer, automated image cytometry of bronchial washings is a sensitive and reliable method for the detection of malignant changes in the tracheobronchial mucosa. The automated procedure seems well suited not only for analysing bronchial washings, but also for a screening procedure. PMID- 10684238 TI - [Irrigation drainage and fibrinolysis for treatment of parapneumonial pleural empyema]. AB - We report on our experiences on 336 patients suffering from manifest pleural empyema within a period of 10 years (1985-1995). Considering the pathogenesis, particularly the results of 218 patients with "parapneumonic pleural empyema" were analysed retrospectively. Definite healing could be achieved by chest tube placement and pleural irrigation in 201 patients (= 92.2%). Other 11 patients finally needed surgical interventions (= 5%). Only 6 patients could not be cured: An indwelling tube was palliatively inserted once and 5 patients died in the course of the medical treatment (mortality = 2.3%). Within the first years the irrigation therapy was performed using a single chest tube (n = 38%) but since 1989 a double-lumen drainage was used (n = 158). Since 1987 in most cases (n = 182) intrapleural medicinal fibrinolysis was performed by instillation of streptokinase (Varidase N). If outward invasively pretreated patients (n = 30) are analysed separately, a statistical dependency can be found between the duration and the way of treatment. Without significant difference between the groups (Gr) the average duration of treatment using a single tube without fibrinolysis (Gr1) was 31.8 days, but 26.5 days using a single tube combined with fibrinolysis (Gr2). A clear shortening of the duration is detectable if patients were treated with a combination of double lumen drainage and fibrinolysis: If 2 tubes were used (Gr4) the treatment lasted 20.6 days, using one double-lumen tube (Gr5) it took 19.8 days. There is a proof of significance at comparison of Gr4 with Gr1 (p = 0.005). Gr5 with Gr1 (p < 0.001) and Gr5 with Gr2 (p = 0.014) respectively. A significant longer duration of treatment (40.6 days, p < 0.001) is found for the group of the pretreated patients, if compared with the corresponding groups Gr4 or Gr5. CONCLUSION: Parapneumonic empyemas most often can be cured by irrigation drainage. The mortality is comparatively low. The shortest duration of treatment is needed using the combination of a double-lumen tube with intrapleural instillation of a fibrinolytic agent (Varidase N). Invasively pretreated patients need significantly longer durations at same form of treatment. PMID- 10684239 TI - [Alveolar proteinosis. Case report and review of a rare disease entity]. PMID- 10684240 TI - [Geographic and economic aspects of inhalation therapy]. PMID- 10684241 TI - [Pulmonary trichomoniasis: diagnosis based on identification of irritation in bronchoalveolar lavage]. AB - Bronchopulmonary infections caused by trichomonads have been reported mainly in patients with pre-existing pulmonary or debilitating disease (e.g. bronchial carcinoma, lung abscess, bronchiectasis). Pulmonary trichomoniasis is most often due to infection with Trichomonas tenax, usually regarded as a harmless commensal of the human mouth, and may rarely be caused by other trichomonas species. A 45 year old female presented with a dry cough, exertional dyspnoea and malaise. These symptoms persisted for 6 months regardless of anti-inflammatory and anti obstructive inhalative therapy. Sarcoidosis of the lungs, diagnosed 20 years prior, had been asymptomatic since and there was no coexistent disease. Laboratory data revealed increased ACE-levels (90 IE/ml) and lung function showed bronchial hyperreactivity on histamine challenge. No other abnormalities were found (chest x-ray, bronchoscopy, lung function test, blood count and serum calcium). The diagnosis was based on the cytological identification of numerous trophozoites of T. tenax in the bronchoalveolar lavage. Therapy with oral metronidazol for 40 days led to complete recovery from symptoms and normalisation of ACE serum levels. The patient has remained well for 12 months since. The pathogenicity of oral trichomonads in the non-immunocompromised host remains uncertain. Our patient had no known medical risk factors by comparison with published cases. The case illustrates the clinical relevance of pulmonary trichomoniasis in an otherwise healthy person. PMID- 10684242 TI - [Abscessed pneumonia caused by Pseudomonas aeruginosa as an occupational disease in a metal driller]. AB - We report the case of a 29-year-old man without immunodeficiency who acquired Pseudomonas aeruginosa pneumonia complicated by pulmonary abscess. The source of infection could be identified as aerosolized metalworking fluid at his workplace contaminated with Pseudomonas aeruginosa. A high titer of specific IgG antibodies (type-III-sensitization, Gell & Coombs) against Pseudomonas aeruginosa has been identified in the patients serum as an indicator for longstanding occupational airborne exposure to contaminated metalworking fluid. This community-acquired pneumonia has been reported to the industrial injuries insurance as an occupational disease for discussion of legal consequences and development of effective measures of prevention. PMID- 10684243 TI - [Asthma self care according to the traffic light principle: which peak flow, which guidelines?]. PMID- 10684244 TI - [Fast-acting combinations of inhalable corticoids and long-acting beta2 sympathomimetics as long-term therapy of bronchial asthma. Opinions of an expert group. R. Buhl et al. Pneumology 53 (1999) 210-212]. PMID- 10684245 TI - Drosophila apoptosis and Bcl-2 genes: outliers fly in. PMID- 10684246 TI - Spatial separation of parental genomes in preimplantation mouse embryos. AB - We have used two different experimental approaches to demonstrate topological separation of parental genomes in preimplantation mouse embryos: mouse eggs fertilized with 5-bromodeoxyuridine (BrdU)-labeled sperm followed by detection of BrdU in early diploid embryos, and differential heterochromatin staining in mouse interspecific hybrid embryos. Separation of chromatin according to parental origin was preserved up to the four-cell embryo stage and then gradually disappeared. In F1 hybrid animals, genome separation was also observed in a proportion of somatic cells. Separate nuclear compartments during preimplantation development, when extreme chromatin remodelling occurs, and possibly in some differentiated cell types, may be associated with epigenetic reprogramming. PMID- 10684247 TI - The yeast nuclear pore complex: composition, architecture, and transport mechanism. AB - An understanding of how the nuclear pore complex (NPC) mediates nucleocytoplasmic exchange requires a comprehensive inventory of the molecular components of the NPC and a knowledge of how each component contributes to the overall structure of this large molecular translocation machine. Therefore, we have taken a comprehensive approach to classify all components of the yeast NPC (nucleoporins). This involved identifying all the proteins present in a highly enriched NPC fraction, determining which of these proteins were nucleoporins, and localizing each nucleoporin within the NPC. Using these data, we present a map of the molecular architecture of the yeast NPC and provide evidence for a Brownian affinity gating mechanism for nucleocytoplasmic transport. PMID- 10684248 TI - Assembly of smooth muscle myosin by the 38k protein, a homologue of a subunit of pre-mRNA splicing factor-2. AB - Smooth muscle myosin in the dephosphorylated state does not form filaments in vitro. However, thick filaments, which are composed of myosin and myosin-binding protein(s), persist in smooth muscle cells, even if myosin is subjected to the phosphorylation- dephosphorylation cycle. The characterization of telokin as a myosin-assembling protein successfully explained the discrepancy. However, smooth muscle cells that are devoid of telokin have been observed. We expected to find another ubiquitous protein with a similar role, and attempted to purify it from chicken gizzard. The 38k protein bound to both phosphorylated and dephosphorylated myosin to a similar extent. The effect of the myosin-binding activity was to assemble dephosphorylated myosin into filaments, although it had no effect on the phosphorylated myosin. The 38k protein bound to myosin with both COOH-terminal 20 and NH(2)-terminal 28 residues of the 38k protein being essential for myosin binding. The amino acid sequence of the 38k protein was not homologous to telokin, but to human p32, which was originally found in nuclei as a subunit of pre-mRNA splicing factor-2. Western blotting showed that the protein was expressed in various smooth muscles. Immunofluorescence microscopy with cultured smooth muscle cells revealed colocalization of the 38k protein with myosin and with other cytoskeletal elements. The absence of nuclear immunostaining was discussed in relation to smooth muscle differentiation. PMID- 10684249 TI - Gelsolin deficiency blocks podosome assembly and produces increased bone mass and strength. AB - Osteoclasts are unique cells that utilize podosomes instead of focal adhesions for matrix attachment and cytoskeletal remodeling during motility. We have shown that osteopontin (OP) binding to the alpha(v)beta(3) integrin of osteoclast podosomes stimulated cytoskeletal reorganization and bone resorption by activating a heteromultimeric signaling complex that includes gelsolin, pp(60c src), and phosphatidylinositol 3'-kinase. Here we demonstrate that gelsolin deficiency blocks podosome assembly and alpha(v)beta(3)-stimulated signaling related to motility in gelsolin-null mice. Gelsolin-deficient osteoclasts were hypomotile due to retarded remodeling of the actin cytoskeleton. They failed to respond to the autocrine factor, OP, with stimulation of motility and bone resorption. Gelsolin deficiency was associated with normal skeletal development and endochondral bone growth. However, gelsolin-null mice had mildly abnormal epiphyseal structure, retained cartilage proteoglycans in metaphyseal trabeculae, and increased trabecular thickness. With age, the gelsolin-deficient mice expressed increased trabecular and cortical bone thickness producing mechanically stronger bones. These observations demonstrate the critical role of gelsolin in podosome assembly, rapid cell movements, and signal transduction through the alpha(v)beta(3) integrin. PMID- 10684250 TI - Regulation of skeletal progenitor differentiation by the BMP and retinoid signaling pathways. AB - The generation of the paraxial skeleton requires that commitment and differentiation of skeletal progenitors is precisely coordinated during limb outgrowth. Several signaling molecules have been identified that are important in specifying the pattern of these skeletal primordia. Very little is known, however, about the mechanisms regulating the differentiation of limb mesenchyme into chondrocytes. Overexpression of RARalpha in transgenic animals interferes with chondrogenesis and leads to appendicular skeletal defects (Cash, D.E., C.B. Bock, K. Schughart, E. Linney, and T.M. Underhill. 1997. J. Cell Biol. 136:445 457). Further analysis of these animals shows that expression of the transgene in chondroprogenitors maintains a prechondrogenic phenotype and prevents chondroblast differentiation even in the presence of BMPs, which are known stimulators of cartilage formation. Moreover, an RAR antagonist accelerates chondroblast differentiation as demonstrated by the emergence of collagen type II expressing cells much earlier than in control or BMP-treated cultures. Addition of Noggin to limb mesenchyme cultures inhibits cartilage formation and the appearance of precartilaginous condensations. In contrast, abrogation of retinoid signaling is sufficient to induce the expression of the chondroblastic phenotype in the presence of Noggin. These findings show that BMP and RAR-signaling pathways appear to operate independently to coordinate skeletal development, and that retinoid signaling can function in a BMP-independent manner to induce cartilage formation. Thus, retinoid signaling appears to play a novel and unexpected role in skeletogenesis by regulating the emergence of chondroblasts from skeletal progenitors. PMID- 10684251 TI - Interaction among GSK-3, GBP, axin, and APC in Xenopus axis specification. AB - Glycogen synthase kinase 3 (GSK-3) is a constitutively active kinase that negatively regulates its substrates, one of which is beta-catenin, a downstream effector of the Wnt signaling pathway that is required for dorsal-ventral axis specification in the Xenopus embryo. GSK-3 activity is regulated through the opposing activities of multiple proteins. Axin, GSK-3, and beta-catenin form a complex that promotes the GSK-3-mediated phosphorylation and subsequent degradation of beta-catenin. Adenomatous polyposis coli (APC) joins the complex and downregulates beta-catenin in mammalian cells, but its role in Xenopus is less clear. In contrast, GBP, which is required for axis formation in Xenopus, binds and inhibits GSK-3. We show here that GSK-3 binding protein (GBP) inhibits GSK-3, in part, by preventing Axin from binding GSK-3. Similarly, we present evidence that a dominant-negative GSK-3 mutant, which causes the same effects as GBP, keeps endogenous GSK-3 from binding to Axin. We show that GBP also functions by preventing the GSK-3-mediated phosphorylation of a protein substrate without eliminating its catalytic activity. Finally, we show that the previously demonstrated axis-inducing property of overexpressed APC is attributable to its ability to stabilize cytoplasmic beta-catenin levels, demonstrating that APC is impinging upon the canonical Wnt pathway in this model system. These results contribute to our growing understanding of how GSK-3 regulation in the early embryo leads to regional differences in beta-catenin levels and establishment of the dorsal axis. PMID- 10684252 TI - Debcl, a proapoptotic Bcl-2 homologue, is a component of the Drosophila melanogaster cell death machinery. AB - Bcl-2 family of proteins are key regulators of apoptosis. Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects. Here, we report the identification and characterization of Debcl, the first Bcl-2 homologue in Drosophila melanogaster. Structurally, Debcl is similar to Bax-like proapoptotic Bcl-2 family members. Ectopic expression of Debcl in cultured cells and in transgenic flies causes apoptosis, which is inhibited by coexpression of the baculovirus caspase inhibitor P35, indicating that Debcl is a proapoptotic protein that functions in a caspase-dependent manner. debcl expression correlates with developmental cell death in specific Drosophila tissues. We also show that debcl genetically interacts with diap1 and dark, and that debcl-mediated apoptosis is not affected by gene dosage of rpr, hid, and grim. Biochemically, Debcl can interact with several mammalian and viral prosurvival Bcl-2 family members, but not with the proapoptotic members, suggesting that it may regulate apoptosis by antagonizing prosurvival Bcl-2 proteins. RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos. These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects. PMID- 10684253 TI - Intracellular distribution of mammalian protein kinase A catalytic subunit altered by conserved Asn2 deamidation. AB - The catalytic (C) subunit of protein kinase A functions both in the cytoplasm and the nucleus. A major charge variant representing about one third of the enzyme in striated muscle results from deamidation in vivo of the Asn2 residue at the conserved NH(2)-terminal sequence myrGly-Asn-Ala (Jedrzejewski, P.T., A. Girod, A. Tholey, N. Konig, S. Thullner, V. Kinzel, and D. Bossemeyer. 1998. Protein Sci. 7:457-469). Because of the increase of electronegativity by generation of Asp2, it is reminiscent of a myristoyl-electrostatic switch. To compare the intracellular distribution of the enzymes, both forms of porcine or bovine heart enzyme were microinjected into the cytoplasm of mouse NIH 3T3 cells after conjugation with fluorescein, rhodamine, or in unlabeled form. The nuclear/cytoplasmic fluorescence ratio (N/C) was analyzed in the presence of cAMP (in the case of unlabeled enzyme by antibodies). Under all circumstances, the N/C ratio obtained with the encoded Asn2 form was significantly higher than that with the deamidated, Asp2 form; i.e., the Asn2 form reached a larger nuclear concentration than the Asp2 form. Comparable data were obtained with a human cell line. The differential intracellular distribution of both enzyme forms is also reflected by functional data. It correlates with the degree of phosphorylation of the key serine in CREB family transcription factors in the nucleus. Microinjection of myristoylated recombinant bovine Calpha and the Asn2 deletion mutant of it yielded N/C ratios in the same range as encoded native enzymes. Thus, Asn2 seems to serve as a potential site for modulating electronegativity. The data indicate that the NH(2)-terminal domain of the PKA C-subunit contributes to the intracellular distribution of free enzyme, which can be altered by site specific in vivo deamidation. The model character for other signaling proteins starting with myrGly-Asn is discussed. PMID- 10684254 TI - Induction of caveolae in the apical plasma membrane of Madin-Darby canine kidney cells. AB - In this paper, we have analyzed the behavior of antibody cross-linked raft associated proteins on the surface of MDCK cells. We observed that cross-linking of membrane proteins gave different results depending on whether cross-linking occurred on the apical or basolateral plasma membrane. Whereas antibody cross linking induced the formation of large clusters on the basolateral membrane, resembling those observed on the surface of fibroblasts (Harder, T., P. Scheiffele, P. Verkade, and K. Simons. 1998. J. Cell Biol. 929-942), only small ( approximately 100 nm) clusters formed on the apical plasma membrane. Cross-linked apical raft proteins e.g., GPI-anchored placental alkaline phosphatase (PLAP), influenza hemagglutinin, and gp114 coclustered and were internalized slowly ( approximately 10% after 60 min). Endocytosis occurred through surface invaginations that corresponded in size to caveolae and were labeled with caveolin-1 antibodies. Upon cholesterol depletion the internalization of PLAP was completely inhibited. In contrast, when a non-raft protein, the mutant LDL receptor LDLR-CT22, was cross-linked, it was excluded from the clusters of raft proteins and was rapidly internalized via clathrin-coated pits. Since caveolae are normally present on the basolateral membrane but lacking from the apical side, our data demonstrate that antibody cross-linking induced the formation of caveolae, which slowly internalized cross-linked clusters of raft-associated proteins. PMID- 10684255 TI - Live Salmonella recruits N-ethylmaleimide-sensitive fusion protein on phagosomal membrane and promotes fusion with early endosome. AB - To understand intracellular trafficking modulations by live Salmonella, we investigated the characteristics of in vitro fusion between endosomes and phagosomes containing live (LSP) or dead Salmonella (DSP). We observed that fusion of both DSP and LSP were time, temperature and cytosol dependent. GTPgammaS and treatment of the phagosomes with Rab-GDI inhibited fusion, indicating involvement of Rab-GTPases. LSP were rich in rab5, alpha-SNAP, and NSF, while DSP mainly contained rab7. Fusion of endosomes with DSP was inhibited by ATP depletion, N-ethylmaleimide (NEM) treatment, and in NEM-sensitive factor (NSF)-depleted cytosol. In contrast, fusion of endosomes with LSP was not inhibited by ATP depletion or NEM treatment, and occurred in NSF-depleted cytosol. However, ATPgammaS inhibited both fusion events. Fusion of NEM-treated LSP with endosomes was abrogated in NSF- depleted cytosol and was restored by adding purified NSF, whereas no fusion occurred with NEM-treated DSP, indicating that NSF recruitment is dependent on continuous signals from live Salmonella. Binding of NSF with LSP required prior presence of rab5 on the phagosome. We have also shown that rab5 specifically binds with Sop E, a protein from Salmonella. Our results indicate that live Salmonella help binding of rab5 on the phagosomes, possibly activate the SNARE which leads to further recruitment of alpha-SNAP for subsequent binding with NSF to promote fusion of the LSP with early endosomes and inhibition of their transport to lysosomes. PMID- 10684256 TI - Exocytotic insertion of calcium channels constrains compensatory endocytosis to sites of exocytosis. AB - Proteins inserted into the cell surface by exocytosis are thought to be retrieved by compensatory endocytosis, suggesting that retrieval requires granule proteins. In sea urchin eggs, calcium influx through P-type calcium channels is required for retrieval, and the large size of sea urchin secretory granules permits the direct observation of retrieval. Here we demonstrate that retrieval is limited to sites of prior exocytosis. We tested whether channel distribution can account for the localization of retrieval at exocytotic sites. We find that P-channels reside on secretory granules before fertilization, and are translocated to the egg surface by exocytosis. Our study provides strong evidence that the transitory insertion of P-type calcium channels in the surface membrane plays an obligatory role in the mechanism coupling exocytosis and compensatory endocytosis. PMID- 10684257 TI - A transmembrane segment determines the steady-state localization of an ion transporting adenosine triphosphatase. AB - The H,K-adenosine triphosphatase (ATPase) of gastric parietal cells is targeted to a regulated membrane compartment that fuses with the apical plasma membrane in response to secretagogue stimulation. Previous work has demonstrated that the alpha subunit of the H, K-ATPase encodes localization information responsible for this pump's apical distribution, whereas the beta subunit carries the signal responsible for the cessation of acid secretion through the retrieval of the pump from the surface to the regulated intracellular compartment. By analyzing the sorting behaviors of a number of chimeric pumps composed of complementary portions of the H, K-ATPase alpha subunit and the highly homologous Na,K-ATPase alpha subunit, we have identified a portion of the gastric H,K-ATPase, which is sufficient to redirect the normally basolateral Na,K-ATPase to the apical surface in transfected epithelial cells. This motif resides within the fourth of the H,K ATPase alpha subunit's ten predicted transmembrane domains. Although interactions with glycosphingolipid-rich membrane domains have been proposed to play an important role in the targeting of several apical membrane proteins, the apically located chimeras are not found in detergent-insoluble complexes, which are typically enriched in glycosphingolipids. Furthermore, a chimera incorporating the Na, K-ATPase alpha subunit fourth transmembrane domain is apically targeted when both of its flanking sequences derive from H,K-ATPase sequence. These results provide the identification of a defined apical localization signal in a polytopic membrane transport protein, and suggest that this signal functions through conformational interactions between the fourth transmembrane spanning segment and its surrounding sequence domains. PMID- 10684258 TI - Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis. AB - E- and N-cadherin are calcium-dependent cell adhesion molecules that mediate cell cell adhesion and also modulate cell migration and tumor invasiveness. The loss of E-cadherin-mediated adhesion has been shown to play an important role in the transition of epithelial tumors from a benign to an invasive state. However, recent evidence indicates that another member of the cadherin family, N-cadherin, is expressed in highly invasive tumor cell lines that lacked E-cadherin expression. These findings have raised the possibility that N-cadherin contributes to the invasive phenotype. To determine whether N-cadherin promotes invasion and metastasis, we transfected a weakly metastatic and E-cadherin expressing breast cancer cell line, MCF-7, with N-cadherin and analyzed the effects on cell migration, invasion, and metastasis. Transfected cells expressed both E- and N-cadherin and exhibited homotypic cell adhesion from both molecules. In vitro, N-cadherin-expressing cells migrated more efficiently, showed an increased invasion of Matrigel, and adhered more efficiently to monolayers of endothelial cells. All cells produced low levels of the matrix metalloproteinase MMP-9, which was dramatically upregulated by treatment with FGF-2 only in N cadherin-expressing cells. Migration and invasion of Matrigel were also greatly enhanced by this treatment. When injected into the mammary fat pad of nude mice, N-cadherin-expressing cells, but not control MCF-7 cells, metastasized widely to the liver, pancreas, salivary gland, omentum, lung, lymph nodes, and lumbar spinal muscle. The expression of both E- and N-cadherin was maintained both in the primary tumors and metastatic lesions. These results demonstrate that N cadherin promotes motility, invasion, and metastasis even in the presence of the normally suppressive E-cadherin. The increase in MMP-9 production by N-cadherin expressing cells in response to a growth factor may endow them with a greater ability to penetrate matrix protein barriers, while the increase in their adherence to endothelium may improve their ability to enter and exit the vasculature, two properties that may be responsible for metastasis of N-cadherin expressing cells. PMID- 10684259 TI - Oncogenic Raf-1 disrupts epithelial tight junctions via downregulation of occludin. AB - Occludin is an integral membrane protein of the epithelial cell tight junction (TJ). Its potential role in coordinating structural and functional events of TJ formation has been suggested recently. Using a rat salivary gland epithelial cell line (Pa-4) as a model system, we have demonstrated that occludin not only is a critical component of functional TJs but also controls the phenotypic changes associated with epithelium oncogenesis. Transfection of an oncogenic Raf-1 into Pa-4 cells resulted in a complete loss of TJ function and the acquisition of a stratified phenotype that lacked cell-cell contact growth control. The expression of occludin and claudin-1 was downregulated, and the distribution patterns of ZO 1 and E-cadherin were altered. Introduction of the human occludin gene into Raf-1 activated Pa-4 cells resulted in reacquisition of a monolayer phenotype and the formation of functionally intact TJs. In addition, the presence of exogenous occludin protein led to a recovery in claudin-1 protein level, relocation of the zonula occludens 1 protein (ZO-1) to the TJ, and redistribution of E-cadherin to the lateral membrane. Furthermore, the expression of occludin inhibited anchorage independent growth of Raf-1-activated Pa-4 cells in soft agarose. Thus, occludin may act as a pivotal signaling molecule in oncogenic Raf- 1-induced disruption of TJs, and regulates phenotypic changes associated with epithelial cell transformation. PMID- 10684260 TI - The small leucine-rich repeat proteoglycan biglycan binds to alpha-dystroglycan and is upregulated in dystrophic muscle. AB - The dystrophin-associated protein complex (DAPC) is necessary for maintaining the integrity of the muscle cell plasma membrane and may also play a role in coordinating signaling events at the cell surface. The alpha-/beta-dystroglycan subcomplex of the DAPC forms a critical link between the cytoskeleton and the extracellular matrix. A ligand blot overlay assay was used to search for novel dystroglycan binding partners in postsynaptic membranes from Torpedo electric organ. An approximately 125-kD dystroglycan-binding polypeptide was purified and shown by peptide microsequencing to be the Torpedo ortholog of the small leucine rich repeat chondroitin sulfate proteoglycan biglycan. Biglycan binding to alpha dystroglycan was confirmed by coimmunoprecipitation with both native and recombinant alpha-dystroglycan. The biglycan binding site was mapped to the COOH terminal third of alpha-dystroglycan. Glycosylation of alpha-dystroglycan is not necessary for this interaction, but binding is dependent upon the chondroitin sulfate side chains of biglycan. In muscle, biglycan is detected at both synaptic and nonsynaptic regions. Finally, biglycan expression is elevated in muscle from the dystrophic mdx mouse. These findings reveal a novel binding partner for alpha dystroglycan and demonstrate a novel avenue for interaction of the DAPC and the extracellular matrix. These results also raise the possibility of a role for biglycan in the pathogenesis, and perhaps the treatment, of muscular dystrophy. PMID- 10684261 TI - Shedding of syndecan-1 and -4 ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive metalloproteinase. AB - The syndecan family of four transmembrane heparan sulfate proteoglycans binds a variety of soluble and insoluble extracellular effectors. Syndecan extracellular domains (ectodomains) can be shed intact by proteolytic cleavage of their core proteins, yielding soluble proteoglycans that retain the binding properties of their cell surface precursors. Shedding is accelerated by PMA activation of protein kinase C, and by ligand activation of the thrombin (G-protein-coupled) and EGF (protein tyrosine kinase) receptors (Subramanian, S.V., M.L. Fitzgerald, and M. Bernfield. 1997. J. Biol. Chem. 272:14713-14720). Syndecan-1 and -4 ectodomains are found in acute dermal wound fluids, where they regulate growth factor activity (Kato, M., H. Wang, V. Kainulainen, M.L. Fitzgerald, S. Ledbetter, D.M. Ornitz, and M. Bernfield. 1998. Nat. Med. 4:691-697) and proteolytic balance (Kainulainen, V., H. Wang, C. Schick, and M. Bernfield. 1998. J. Biol. Chem. 273:11563-11569). However, little is known about how syndecan ectodomain shedding is regulated. To elucidate the mechanisms that regulate syndecan shedding, we analyzed several features of the process that sheds the syndecan-1 and -4 ectodomains. We find that shedding accelerated by various physiologic agents involves activation of distinct intracellular signaling pathways; and the proteolytic activity responsible for cleavage of syndecan core proteins, which is associated with the cell surface, can act on unstimulated adjacent cells, and is specifically inhibited by TIMP-3, a matrix-associated metalloproteinase inhibitor. In addition, we find that the syndecan-1 core protein is cleaved on the cell surface at a juxtamembrane site; and the proteolytic activity responsible for accelerated shedding differs from that involved in constitutive shedding of the syndecan ectodomains. These results demonstrate the existence of highly regulated mechanisms that can rapidly convert syndecans from cell surface receptors or coreceptors to soluble heparan sulfate proteoglycan effectors. Because the shed ectodomains are found and function in vivo, regulation of syndecan ectodomain shedding by physiological mediators indicates that shedding is a response to specific developmental and pathophysiological cues. PMID- 10684262 TI - Tissue transglutaminase is an integrin-binding adhesion coreceptor for fibronectin. AB - The protein cross-linking enzyme tissue transglutaminase binds in vitro with high affinity to fibronectin via its 42-kD gelatin-binding domain. Here we report that cell surface transglutaminase mediates adhesion and spreading of cells on the 42 kD fibronectin fragment, which lacks integrin-binding motifs. Overexpression of tissue transglutaminase increases its amount on the cell surface, enhances adhesion and spreading on fibronectin and its 42-kD fragment, enlarges focal adhesions, and amplifies adhesion-dependent phosphorylation of focal adhesion kinase. These effects are specific for tissue transglutaminase and are not shared by its functional homologue, a catalytic subunit of factor XIII. Adhesive function of tissue transglutaminase does not require its cross-linking activity but depends on its stable noncovalent association with integrins. Transglutaminase interacts directly with multiple integrins of beta1 and beta3 subfamilies, but not with beta2 integrins. Complexes of transglutaminase with integrins are formed inside the cell during biosynthesis and accumulate on the surface and in focal adhesions. Together our results demonstrate that tissue transglutaminase mediates the interaction of integrins with fibronectin, thereby acting as an integrin-associated coreceptor to promote cell adhesion and spreading. PMID- 10684263 TI - Nuclear matrix attachment regions of human papillomavirus type 16 repress or activate the E6 promoter, depending on the physical state of the viral DNA. AB - Two nuclear matrix attachment regions (MARs) bracket a 550-bp segment of the long control region (LCR) containing the epithelial cell-specific enhancer and the E6 promoter of human papillomavirus type 16 (HPV-16). One of these MARs is located in the 5' third of the LCR (5'-LCR-MAR); the other lies within the E6 gene (E6 MAR). To study their function, we linked these MARs in various natural or artificial permutations to a chimeric gene consisting of the HPV-16 enhancer promoter segment and a reporter gene. In transient transfections of HeLa cells, the presence of either of these two MARs strongly represses reporter gene expression. In contrast to this, but similar to the published behavior of cellular MARs, reporter gene expression is stimulated strongly by the E6-MAR and moderately by the 5'-LCR-MAR in stable transfectants of HeLa or C33A cells. To search for binding sites of soluble nuclear proteins which may be responsible for repression during transient transfections, we performed electrophoretic mobility shift assays (EMSAs) of overlapping oligonucleotides that represented all sequences of these two MARs. Both MARs contain multiple sites for two strongly binding proteins and weak binding sites for additional factors. The strongest complex, with at least five binding sites in each MAR, is generated by the CCAAT displacement factor (CDP)/Cut, as judged by biochemical purification, by EMSAs with competing oligonucleotides and with anti-CDP/Cut oligonucleotides, and by mutations. CDP/Cut, a repressor that is down-regulated during differentiation, apparently represses HPV-16 transcription in undifferentiated epithelials cells and in HeLa cells, which are rich in CDP/Cut. In analogy to poorly understood mechanisms acting on cellular MARs, activation after physical linkage to chromosomal DNA may result from competition between the nuclear matrix and CDP/Cut. Our observations show that cis-responsive elements that regulate the HPV 16 E6 promoter are tightly clustered over at least 1.3 kb and occur throughout the E6 gene. HPV-16 MARs are context dependent transcriptional enhancers, and activated expression of HPV-16 oncogenes dependent on chromosomal integration may positively select tumorigenic cells during the multistep etiology of cervical cancer. PMID- 10684264 TI - Immunization with a modified vaccinia virus expressing simian immunodeficiency virus (SIV) Gag-Pol primes for an anamnestic Gag-specific cytotoxic T-lymphocyte response and is associated with reduction of viremia after SIV challenge. AB - The immunogenicity and protective efficacy of a modified vaccinia virus Ankara (MVA) recombinant expressing the simian immunodeficiency virus (SIV) Gag-Pol proteins (MVA-gag-pol) was explored in rhesus monkeys expressing the major histocompatibility complex (MHC) class I allele, MamuA*01. Macaques received four sequential intramuscular immunizations with the MVA-gag-pol recombinant virus or nonrecombinant MVA as a control. Gag-specific cytotoxic T-lymphocyte (CTL) responses were detected in all MVA-gag-pol-immunized macaques by both functional assays and flow cytometric analyses of CD8(+) T cells that bound a specific MHC complex class I-peptide tetramer, with levels peaking after the second immunization. Following challenge with uncloned SIVsmE660, all macaques became infected; however, viral load set points were lower in MVA-gag-pol-immunized macaques than in the MVA-immunized control macaques. MVA-gag-pol-immunized macaques exhibited a rapid and substantial anamnestic CTL response specific for the p11C, C-M Gag epitope. The level at which CTL stabilized after resolution of primary viremia correlated inversely with plasma viral load set point (P = 0.03). Most importantly, the magnitude of reduction in viremia in the vaccinees was predicted by the magnitude of the vaccine-elicited CTL response prior to SIV challenge. PMID- 10684265 TI - Covalent modification of the transactivator protein IE2-p86 of human cytomegalovirus by conjugation to the ubiquitin-homologous proteins SUMO-1 and hSMT3b. AB - The 86-kDa IE2 protein (IE2-p86) of human cytomegalovirus (HCMV) is a potent transactivator of viral as well as cellular promoters. Several lines of evidence indicate that this broad transactivation spectrum is mediated by protein-protein interactions. To identify novel cellular binding partners, we performed a yeast two-hybrid screen using a N-terminal deletion mutant of IE2-p86 comprising amino acids 135 to 579 as a bait. Here, we report the isolation of two ubiquitin homologous proteins, SUMO-1 and hSMT3b, as well as their conjugating activity hUBC9 (human ubiquitin-conjugating enzyme 9) as specific interaction partners of HCMV IE2. The polypeptides SUMO-1 and hSMT3b have previously been shown to be covalently coupled to a subset of nuclear proteins such as the nuclear domain 10 (ND10) proteins PML and Sp100 in a manner analogous to ubiquitinylation, which we call SUMOylation. By Western blot analysis, we were able to show that the IE2-p86 protein can be partially converted to a 105-kDa isoform in a dose-dependent manner after cotransfection of an epitope-tagged SUMO-1. Immunoprecipitation experiments of the conjugated isoforms using denaturing conditions further confirmed the covalent coupling of SUMO-1 or hSMT3b to IE2-p86 both after transient transfection and after lytic infection of human primary fibroblasts. Moreover, we defined two modification sites within IE2, located in an immediate vicinity at amino acid positions 175 and 180, which appear to be used alternatively for coupling. By using a SUMOylation-defective mutant, we showed that the targeting of IE2-p86 to ND10 occurs independent of this modification. However, a strong reduction of IE2-mediated transactivation of two viral early promoters and a heterologous promoter was observed in cotransfection analysis with the SUMOylation-defective mutant. This suggests a functional relevance of covalent modification by ubiquitin-homologous proteins for IE2-mediated transactivation, possibly by providing an additional interaction motif for cellular cofactors. PMID- 10684266 TI - Phylogenetic analyses indicate an atypical nurse-to-patient transmission of human immunodeficiency virus type 1. AB - A human immunodeficiency virus (HIV)-negative patient with no risk factor experienced HIV type 1 (HIV-1) primary infection 4 weeks after being hospitalized for surgery. Among the medical staff, only two night shift nurses were identified as HIV-1 seropositive. No exposure to blood was evidenced. To test the hypothesis of a possible nurse-to-patient transmission, phylogenetic analyses were conducted using two HIV-1 genomic regions (pol reverse transcriptase [RT] and env C2C4), each compared with reference strains and large local control sets (57 RT and 41 C2C4 local controls). Extensive analyses using multiple methodologies allowed us to test the robustness of phylogeny inference and to assess transmission hypotheses. Results allow us to unambiguously exclude one HIV-positive nurse and strongly suggest the other HIV-positive nurse as the source of infection of the patient. PMID- 10684267 TI - Feline immunodeficiency virus Vif localizes to the nucleus. AB - Monoclonal antibodies prepared against recombinant Vif derived from the 34TF10 strain of feline immunodeficiency virus (FIV) were used to assess the expression and localization of Vif in virus-infected cells. Analyses by Western blotting and by immunoprecipitation from cells infected with FIV-34TF10 revealed the presence of a single 29-kDa species specific for virus-infected cells. Confirmation of antibody specificity was also performed by specific immunoprecipitation of in vitro-transcribed and -translated recombinant Vif. Localization experiments were also performed on virus-infected cells, using different fixation procedures. Results for methanol fixation protocols similar to those reported for localization of human immunodeficiency virus (HIV) Vif showed a predominant cytoplasmic localization for FIV Vif, very similar to localization of HIV type 1 Vif and virtually identical to the localization observed for the Gag antigens of the virus. However, with milder fixation procedures that used 2% formaldehyde at 4 degrees C, FIV Vif was strongly evident in the nucleus. The localization was distinct from the nuclear localization noted with Rev and did not involve the nucleolus. Attempts to show colocalization or coprecipitation of Vif with Gag antigens were unsuccessful. In addition, Vif was not detected in purified FIV virions. The results are consistent with the notion that the primary role of Vif in virus infection initiates in the nucleus. PMID- 10684268 TI - Evolutionary rate and genetic drift of hepatitis C virus are not correlated with the host immune response: studies of infected donor-recipient clusters. AB - Six donor-recipient clusters of hepatitis C virus (HCV)-infected individuals were studied. For five clusters the period of infection of the donor could be estimated, and for all six clusters the time of infection of the recipients from the donor via blood transfusion was also precisely known. Detailed phylogenetic analyses were carried out to investigate the genomic evolution of the viral quasispecies within infected individuals in each cluster. The molecular clock analysis showed that HCV quasispecies within a patient are evolving at the same rate and that donors that have been infected for longer time tend to have a lower evolutionary rate. Phylogenetic analysis based on the split decomposition method revealed different evolutionary patterns in different donor-recipient clusters. Reactivity of antibody against the first hypervariable region (HVR1) of HCV in donor and recipient sera was evaluated and correlated to the calculated evolutionary rate. Results indicate that anti-HVR1 reactivity was related more to the overall level of humoral immune response of the host than to the HVR1 sequence itself, suggesting that the particular sequence of the HVR1 peptides is not the determinant of reactivity. Moreover, no correlation was found between the evolutionary rate or the heterogeneity of the viral quasispecies in the patients and the strength of the immune response to HVR1 epitopes. Rather, the results seem to imply that genetic drift is less dependent on immune pressure than on the rate of evolution and that the genetic drift of HCV is independent of the host immune pressure. PMID- 10684269 TI - The Epstein-Barr virus pol catalytic subunit physically interacts with the BBLF4 BSLF1-BBLF2/3 complex. AB - The Epstein-Barr virus (EBV)-encoded replication proteins that account for the basic reactions at the replication fork are thought to be the EBV Pol holoenzyme, consisting of the BALF5 Pol catalytic and the BMRF1 Pol accessory subunits, the putative helicase-primase complex, comprising the BBLF4, BSLF1, and BBLF2/3 proteins, and the BALF2 single-stranded DNA-binding protein. Immunoprecipitation analyses using anti-BSLF1 or anti-BBLF2/3 protein-specific antibody with clarified lysates of B95-8 cells in a viral productive cycle suggested that the EBV Pol holoenzyme physically interacts with the BBLF4-BSLF1-BBLF2/3 complex to form a large complex. Although the complex was stable in 500 mM NaCl and 1% NP 40, the BALF5 protein became dissociated in the presence of 0.1% sodium dodecyl sulfate. Experiments using lysates from insect cells superinfected with combinations of recombinant baculoviruses capable of expressing each of viral replication proteins showed that not the BMRF1 Pol accessory subunit but rather the BALF5 Pol catalytic subunit directly interacts with the BBLF4-BSLF1-BBLF2/3 complex. Furthermore, double infection with pairs of recombinant viruses revealed that each component of the BBLF4-BSLF1-BBLF2/3 complex makes contact with the BALF5 Pol catalytic subunit. The interactions of the EBV DNA polymerase with the EBV putative helicase-primase complex warrant particular attention because they are thought to coordinate leading- and lagging-strand DNA synthesis at the replication fork. PMID- 10684270 TI - Inhibition of CD3/CD28-mediated activation of the MEK/ERK signaling pathway represses replication of X4 but not R5 human immunodeficiency virus type 1 in peripheral blood CD4(+) T lymphocytes. AB - Binding of human immunodeficiency virus type 1 (HIV-1) to CD4 receptors induces multiple cellular signaling pathways, including the MEK/ERK cascade. While the interaction of X4 HIV-1 with CXCR4 does not seem to activate this pathway, viruses using CCR5 for entry efficiently activate MEK/ERK kinases (W. Popik, J. E. Hesselgesser, and P. M. Pitha, J. Virol. 72:6406-6413, 1998; W. Popik and P. M. Pitha, Virology 252:210-217, 1998). Since the importance of MEK/ERK in the initial steps of viral replication is poorly understood, we have examined the role of MEK/ERK signaling in the CD3- and CD28 (CD3/CD28)-mediated activation of HIV-1 replication in resting peripheral blood CD4(+) T lymphocytes infected with X4 or R5 HIV-1. We have found that the MEK/ERK inhibitor U0126 selectively inhibited CD3/CD28-stimulated replication of X4 HIV-1, while it did not affect the replication of R5 HIV-1. Inhibition of the CD3/CD28-stimulated MEK/ERK pathway did not affect the formation of the early proviral transcripts in cells infected with either X4 or R5 HIV-1, indicating that virus reverse transcription is not affected in the absence of MEK/ERK signaling. In contrast, the levels of nuclear provirus in cells infected with X4 HIV-1, detected by the formation of circular proviral DNA, was significantly lower in cells stimulated in the presence of MEK/ERK inhibitor than in the absence of the inhibitor. However, in cells infected with R5 HIV-1, the inhibition of the MEK/ERK pathway did not affect nuclear localization of the proviral DNA. These data suggest that the nuclear import of X4, but not R5, HIV-1 is dependent on a CD3/CD28-stimulated MEK/ERK pathway. PMID- 10684271 TI - Efficient gene transfer into human CD34(+) cells by a retargeted adenovirus vector. AB - Efficient infection with adenovirus (Ad) vectors based on serotype 5 (Ad5) requires the presence of coxsackievirus-adenovirus receptors (CAR) and alpha(v) integrins on cells. The paucity of these cellular receptors is thought to be a limiting factor for Ad gene transfer into hematopoietic stem cells. In a systematic approach, we screened different Ad serotypes for interaction with noncycling human CD34(+) cells and K562 cells on the level of virus attachment, internalization, and replication. From these studies, serotype 35 emerged as the variant with the highest tropism for CD34(+) cells. A chimeric vector (Ad5GFP/F35) was generated which contained the short-shafted Ad35 fiber incorporated into an Ad5 capsid. This substitution was sufficient to transplant all infection properties from Ad35 to the chimeric vector. The retargeted, chimeric vector attached to a receptor different from CAR and entered cells by an alpha(v) integrin-independent pathway. In transduction studies, Ad5GFP/F35 expressed green fluorescent protein (GFP) in 54% of CD34(+) cells. In comparison, the standard Ad5GFP vector conferred GFP expression to only 25% of CD34(+) cells. Importantly, Ad5GFP transduction, but not Ad5GFP/F35, was restricted to a specific subset of CD34(+) cells expressing alpha(v) integrins. The actual transduction efficiency was even higher than 50% because Ad5GFP/F35 viral genomes were found in GFP-negative CD34(+) cell fractions, indicating that the cytomegalovirus promoter used for transgene expression was not active in all transduced cells. The chimeric vector allowed for gene transfer into a broader spectrum of CD34(+) cells, including subsets with potential stem cell capacity. Fifty-five percent of CD34(+) c-Kit(+) cells expressed GFP after infection with Ad5GFP/F35, whereas only 13% of CD34(+) c-Kit(+) cells were GFP positive after infection with Ad5GFP. These findings represent the basis for studies aimed toward stable gene transfer into hematopoietic stem cells. PMID- 10684272 TI - Containment of simian immunodeficiency virus infection: cellular immune responses and protection from rechallenge following transient postinoculation antiretroviral treatment. AB - To better understand the viral and host factors involved in the establishment of persistent productive infection by primate lentiviruses, we varied the time of initiation and duration of postinoculation antiretroviral treatment with tenofovir (9-[2-(R)-(phosphonomethoxy)propyl]adenine) while performing intensive virologic and immunologic monitoring in rhesus macaques, inoculated intravenously with simian immunodeficiency virus SIVsmE660. Postinoculation treatment did not block the initial infection, but we identified treatment regimens that prevented the establishment of persistent productive infection, as judged by the absence of measurable plasma viremia following drug discontinuation. While immune responses were heterogeneous, animals in which treatment resulted in prevention of persistent productive infection showed a higher frequency and higher levels of SIV-specific lymphocyte proliferative responses during the treatment period compared to control animals, despite the absence of either detectable plasma viremia or seroconversion. Animals protected from the initial establishment of persistent productive infection were also relatively or completely protected from subsequent homologous rechallenge. Even postinoculation treatment regimens that did not prevent establishment of persistent infection resulted in downmodulation of the level of plasma viremia following treatment cessation, compared to the viremia seen in untreated control animals, animals treated with regimens known to be ineffective, or the cumulative experience with the natural history of plasma viremia following infection with SIVsmE660. The results suggest that the host may be able to effectively control SIV infection if the initial exposure occurs under favorable conditions of low viral burden and in the absence of ongoing high level cytopathic infection of responding cells. These findings may be particularly important in relation to prospects for control of primate lentiviruses in the settings of both prophylactic and therapeutic vaccination for prevention of AIDS. PMID- 10684273 TI - Partial rescue of the Vif-negative phenotype of mutant human immunodeficiency virus type 1 strains from nonpermissive cells by intravirion reverse transcription. AB - Virion infectivity factor (Vif) is a protein encoded by human immunodeficiency virus type I (HIV-1) and is essential for viral replication. It appears that Vif functions in the virus-producing cells and affects viral assembly. Viruses with defects in the vif gene (vif-) generated from the "nonpermissive cells" are not able to complete reverse transcription. In previous studies, it was demonstrated that defects in the vif gene also affect endogenous reverse transcription (ERT) when mild detergents were utilized to permeabilize the viral envelope. In this report, we demonstrate that defects in the vif gene have much less of an effect on ERT if detergent is not used. When ERT was driven by addition of deoxyribonucleoside triphosphates (dNTPs) at high concentrations, certain levels of plus-strand viral DNA could also be achieved. Interestingly, if vif- viruses, generated from nonpermissive cells and harboring large quantities of viral DNA generated by ERT, were allowed to infect permissive cells, they could partially bypass the block at intracellular reverse transcription, through which vif- viruses without dNTP treatment could not pass. Consequently, viral infectivity can be partially rescued from the vif- phenotype. Based on our observations, we suggest that vif defects may cause the reverse transcription complex (RT complex) to become sensitive to mild detergent treatments within HIV-1 virions and become unstable in the target cells, such that the process of reverse transcription cannot be efficiently supported. Further dissection of RT complexes of vif- viruses may be key to uncovering the molecular mechanism(s) of Vif in HIV-1 pathogenesis. PMID- 10684274 TI - Rinderpest viruses lacking the C and V proteins show specific defects in growth and transcription of viral RNAs. AB - Rinderpest virus is a morbillivirus and the causative agent of an important disease of cattle and wild bovids. The P genes of all morbilliviruses give rise to two proteins in addition to the P protein itself: use of an alternate start translation site, in a second open reading frame, gives rise to the C protein, while cotranscriptional insertion of an extra base gives rise to the V protein, a fusion of the amino-terminal half of P to a short, highly conserved, cysteine rich zinc binding domain. Little is known about the function of either of these two proteins in the rinderpest virus life cycle. We have constructed recombinant rinderpest viruses in which the expression of these proteins has been suppressed, individually and together, and studied the replication of these viruses in tissue culture. We show that the absence of the V protein has little effect on the replication rate of the virus but does lead to an increase in synthesis of genome and antigenome RNAs and a change in cytopathic effect to a more syncytium-forming phenotype. Virus that does not express the C protein, on the other hand, is clearly defective in growth in all cell lines tested, and this defect appears to be related to a decreased transcription of mRNA from viral genes. The phenotypes of both individual mutant virus types are both expressed in the double mutant expressing neither V nor C. PMID- 10684275 TI - Infection of primary human monocytes by Epstein-Barr virus. AB - Previous studies have reported that infection of monocytes by viruses such as cytomegalovirus and human immunodeficiency virus weakens host natural immunity. In the present study, we demonstrated the capability of Epstein-Barr virus (EBV) to infect and replicate in freshly isolated human monocytes. Using electron microscopy analysis, we observed the presence of EBV virions in the cytoplasm and nuclei of approximately 20% of monocytes. This was confirmed by Southern blot analysis of EBV genomic DNA sequences in isolated nuclei from monocytes. Infection of monocytes by EBV leads to the activation of the replicative cycle. This was supported by the detection of immediate-early lytic mRNA BZLF-1 transcripts, and by the presence of two early lytic transcripts (BALF-2, which appears to function in DNA replication, and BHRF-1, also associated with the replicative cycle). The late lytic BcLF-1 transcripts, which code for the major nucleocapsid protein, were also detected, as well as EBNA-1 transcripts. However, attempts to detect EBNA-2 transcripts have yielded negative results. Viral replication was also confirmed by the release of newly synthesized infectious viral particles in supernatants of EBV-infected monocytes. EBV-infected monocytes were found to have significantly reduced phagocytic activity, as evaluated by the quantification of ingested carboxylated fluoresceinated latex beads. Taken together, our results suggest that EBV infection of monocytes and alteration of their biological functions might represent a new mechanism to disrupt the immune response and promote viral propagation during the early stages of infection. PMID- 10684276 TI - Immune responses following neonatal DNA vaccination are long-lived, abundant, and qualitatively similar to those induced by conventional immunization. AB - Virus infections are devastating to neonates, and the induction of active antiviral immunity in this age group is an important goal. Here, we show that a single neonatal DNA vaccination induces cellular and humoral immune responses which are maintained for a significant part of the animal's life span. We employ a sensitive technique which permits the first demonstration and quantitation, directly ex vivo, of virus-specific CD8(+) T cells induced by DNA immunization. One year postvaccination, antigen-specific CD8(+) T cells were readily detectable and constituted 0.5 to 1% of all CD8(+) T cells. By several criteria-including cytokine production, perforin content, development of lytic ability, and protective capacity-DNA vaccine-induced CD8(+) memory T cells were indistinguishable from memory cells induced by immunization with a conventional (live-virus) vaccine. Analyses of long-term humoral immune responses revealed that, in contrast to the strong immunoglobulin G2a (IgG2a) skewing of the humoral response seen after conventional vaccination, IgG1 and IgG2a levels were similar in DNA-vaccinated neonatal and adult animals, indicating a balanced T helper response. Collectively, these results show that a single DNA vaccination within hours or days of birth can induce long-lasting CD8(+) T- and B-cell responses; there is no need for secondary immunization (boosting). Furthermore, the observed immune responses induced in neonates and in adults are indistinguishable by several criteria, including protection against virus challenge. PMID- 10684277 TI - Increased expression and immunogenicity of sequence-modified human immunodeficiency virus type 1 gag gene. AB - A major challenge for the next generation of human immunodeficiency virus (HIV) vaccines is the induction of potent, broad, and durable cellular immune responses. The structural protein Gag is highly conserved among the HIV type 1 (HIV-1) gene products and is believed to be an important target for the host cell mediated immune control of the virus during natural infection. Expression of Gag proteins for vaccines has been hampered by the fact that its expression is dependent on the HIV Rev protein and the Rev-responsive element, the latter located on the env transcript. Moreover, the HIV genome employs suboptimal codon usage, which further contributes to the low expression efficiency of viral proteins. In order to achieve high-level Rev-independent expression of the Gag protein, the sequences encoding HIV-1(SF2) p55(Gag) were modified extensively. First, the viral codons were changed to conform to the codon usage of highly expressed human genes, and second, the residual inhibitory sequences were removed. The resulting modified gag gene showed increases in p55(Gag) protein expression to levels that ranged from 322- to 966-fold greater than that for the native gene after transient expression of 293 cells. Additional constructs that contained the modified gag in combination with modified protease coding sequences were made, and these showed high-level Rev-independent expression of p55(Gag) and its cleavage products. Density gradient analysis and electron microscopy further demonstrated that the modified gag and gag protease genes efficiently expressed particles with the density and morphology expected for HIV virus-like particles. Mice immunized with DNA plasmids containing the modified gag showed Gag-specific antibody and CD8(+) cytotoxic T-lymphocyte (CTL) responses that were inducible at doses of input DNA 100-fold lower than those associated with plasmids containing the native gag gene. Most importantly, four of four rhesus monkeys that received two or three immunizations with modified gag plasmid DNA demonstrated substantial Gag-specific CTL responses. These results highlight the useful application of modified gag expression cassettes for increasing the potency of DNA and other gene delivery vaccine approaches against HIV. PMID- 10684278 TI - Genetic studies with the fission yeast Schizosaccharomyces pombe suggest involvement of wee1, ppa2, and rad24 in induction of cell cycle arrest by human immunodeficiency virus type 1 Vpr. AB - Accessory protein Vpr of human immunodeficiency virus type 1 (HIV-1) arrests cell cycling at G(2)/M phase in human and simian cells. Recently, it has been shown that Vpr also causes cell cycle arrest in the fission yeast Schizosaccharomyces pombe, which shares the cell cycle regulatory mechanisms with higher eukaryotes including humans. In this study, in order to identify host cellular factors involved in Vpr-induced cell cycle arrest, the ability of Vpr to cause elongated cellular morphology (cdc phenotype) typical of G(2)/M cell cycle arrest in wild type and various mutant strains of S. pombe was examined. Our results indicated that Vpr caused the cdc phenotype in wild-type S. pombe as well as in strains carrying mutations, such as the cdc2-3w, Deltacdc25, rad1-1, Deltachk1, Deltamik1, and Deltappa1 strains. However, other mutants, such as the cdc2-1w, Deltawee1, Deltappa2, and Deltarad24 strains, failed to show a distinct cdc phenotype in response to Vpr expression. Results of these genetic studies suggested that Wee1, Ppa2, and Rad24 might be required for induction of cell cycle arrest by HIV-1 Vpr. Cell proliferation was inhibited by Vpr expression in all of the strains examined including the ones that did not show the cdc phenotype. The results supported the previously suggested possibility that Vpr affects the cell cycle and cell proliferation through different pathways. PMID- 10684279 TI - Mapping in solution shows the peach latent mosaic viroid to possess a new pseudoknot in a complex, branched secondary structure. AB - We have investigated the secondary structure of peach latent mosaic viroid (PLMVd) in solution, and we present here the first description of the structure of a branched viroid in solution. Different PLMVd transcripts of plus polarity were produced by using the circularly permuted RNA method and the exploitation of RNA internal secondary structure to position the 5' and 3' termini and studied by nuclease mapping and binding shift assays using DNA and RNA oligonucleotides. We show that PLMVd folds into a complex, branched secondary structure. In general, this structure is similar to that reported previously, which was based on sequence comparison and computer modelling. The structural microheterogeneity is apparently limited to only some small domains. More importantly, this structure includes a novel pseudoknot that is conserved in all PLMVd isolates and seems to allow folding into a very compact form. This pseudoknot is also found in chrysanthemum chlorotic mottle viroid, suggesting that it is a unique feature of the viroid members of the PLMVd subgroup. PMID- 10684280 TI - Human T-cell leukemia virus type 2 tax mutants that selectively abrogate NFkappaB or CREB/ATF activation fail to transform primary human T cells. AB - Human T-cell leukemia virus (HTLV) Tax protein has been implicated in the HTLV oncogenic process, primarily due to its pleiotropic effects on cellular genes involved in growth regulation and cell cycle control. To date, several approaches attempting to correlate Tax activation of the CREB/activating transcription factor (ATF) or NFkappaB/Rel transcriptional activation pathway to cellular transformation have yielded conflicting results. In this study, we use a unique HTLV-2 provirus (HTLV(c-enh)) that replicates by a Tax-independent mechanism to directly assess the role of Tax transactivation in HTLV-mediated T-lymphocyte transformation. A panel of well-characterized tax-2 mutations is utilized to correlate the respective roles of the CREB/ATF or NFkappaB/Rel signaling pathway. Our results demonstrate that viruses expressing tax-2 mutations that selectively abrogate NFkappaB/Rel or CREB/ATF activation display distinct phenotypes but ultimately fail to transform primary human T lymphocytes. One conclusion consistent with our results is that the activation of NFkappaB/Rel provides a critical proliferative signal early in the cellular transformation process, whereas CREB/ATF activation is required to promote the fully transformed state. However, complete understanding will require correlation of Tax domains important in cellular transformation to those Tax domains important in the modulation of gene transcription, cell cycle control, induction of DNA damage, and other undefined activities. PMID- 10684281 TI - Adaptive mutations in Sindbis virus E2 and Ross River virus E1 that allow efficient budding of chimeric viruses. AB - Alphavirus glycoproteins E2 and E1 form a heterodimer that is required for virus assembly. We have studied adaptive mutations in E2 of Sindbis virus (SIN) and E1 of Ross River virus (RR) that allow these two glycoproteins to interact more efficiently in a chimeric virus that has SIN E2 but RR E1. These mutations include K129E, K131E, and V237F in SIN E2 and S310F and C433R in RR E1. Although RR E1 and SIN E2 will form a chimeric heterodimer, the chimeric virus is almost nonviable, producing about 10(-7) as much virus as SIN at 24 h and 10(-5) as much after 48 h. Chimeras containing one adaptive change produced 3 to 20 times more virus than did the parental chimera, whereas chimeras with two changes produced 10 to 100 times more virus and chimeras containing three mutations produced yields that were 180 to 250 times better. None of the mutations had significant effects upon the parental wild-type viruses, however. Passage of the triple variants eight or nine times resulted in variants that produced virus rapidly and were capable of producing >10(8) PFU/ml of culture fluid within 24 h. These further-adapted variants possessed one or two additional mutations, including E2 V116K, E2-S110N, or E1-T65S. The RR E1-C433R mutation was studied in more detail. This Cys is located in the putative transmembrane domain of E1 and was shown to be palmitoylated. Mutation to Arg-433 resulted in loss of palmitoylation of E1. The positively charged arginine residue within the putative transmembrane domain of E1 would be expected to alter the conformation of this domain. These results suggest that interactions within the transmembrane region are important for the assembly of the E1/E2 heterodimer, as are regions of the ectodomains possibly identified by the locations of adaptive mutations in these regions. Further, the finding that four or five changes in the chimera allow virus production that approaches the levels seen with the parental SIN and exceeds that of the parental RR illustrates that the structure and function of SIN and RR E1s have been conserved during the 50% divergence in sequence that has occurred. PMID- 10684282 TI - Human beta interferon scaffold attachment region inhibits de novo methylation and confers long-term, copy number-dependent expression to a retroviral vector. AB - Moloney murine leukemia virus-based retroviral vector expression is gradually lost during prolonged in vitro culture of CEMSS T cells. However, when the human beta interferon scaffold attachment region (IFN-SAR) was inserted into the vector immediately upstream of the 3' long terminal repeat (LTR), expression was maintained for the length of the study (4 months). Clonal analysis of the retrovirus vector-infected CEMSS cells showed that SAR-containing retroviral vector expression levels were positively correlated with the proviral copy numbers (P < 0.0001), while there was no correlation between the proviral copy numbers and expression levels in control vector-infected clones. Thirty-three percent of the CEMSS cell clones infected with the control vector showed evidence of partial or complete methylation in the 5' LTR region. In sharp contrast, we detected no methylation in the clones infected with the SAR-containing vector. To demonstrate a direct inhibitory effect of methylation on retroviral vector expression, we have transfected 293 cells with in vitro-methylated proviral DNA. In transiently transfected cells, expression of methylated LTR was reduced but not completely inhibited, irrespective of the presence of the IFN-SAR sequence. In stably transfected cells, however, methylation completely abolished expression of the control vector but not of the SAR-containing vector. Furthermore, the expression of the SAR-containing vector was stable over time, indicating the ability of the SAR sequence to alleviate methylation-mediated transcriptional repression of a vector. This study extends our understanding of the mechanisms of retroviral vector inactivation by methylation and provides insight into a functional role for the SAR elements. PMID- 10684283 TI - Repression of the integrated papillomavirus E6/E7 promoter is required for growth suppression of cervical cancer cells. AB - The human papillomavirus (HPV) E2 protein is an important regulator of viral E6 and E7 gene expression. E2 can repress the viral promoter for E6 and E7 expression as well as block progression of the cell cycle in cancer cells harboring the DNA of "high-risk" HPV types. Although the phenomenon of E2 mediated growth arrest of HeLa cells and other HPV-positive cancer cells has been well documented, the specific mechanism by which E2 affects cellular proliferation has not yet been elucidated. Here, we show that bovine papillomavirus (BPV) E2-induced growth arrest of HeLa cells requires the repression of the E6 and E7 promoter. This repression is specific for E2TA and not E2TR, a BPV E2 variant that lacks the N-terminal transactivation domain. We demonstrate that expression of HPV16 E6 and E7 from a heterologous promoter that is not regulated by E2 rescues HeLa cells from E2-mediated growth arrest. Our data indicate that the pathway of E2-mediated growth arrest of HeLa cells requires repression of E6 and E7 expression through an activity specified by the transactivation domain of E2TA. PMID- 10684284 TI - Interaction of the adenovirus IVa2 protein with viral packaging sequences. AB - We have demonstrated previously that the adenovirus L1 52/55-kDa protein binds to the viral IVa2 protein in infected cells. The significance of this interaction was unclear, however, based on the known functions of these two proteins: the 52/55-kDa protein is required for viral DNA packaging, while the IVa2 protein is a transactivator of the major late promoter (MLP). In this report, we have attempted to elucidate a role for each of the two proteins in the other's known function. There is no apparent effect of the 52/55-kDa protein on the interaction of the IVa2 protein with the MLP. Surprisingly, however, we found that the IVa2 protein can interact with the adenoviral packaging signal and that this interaction involves DNA sequences that have previously been demonstrated to be required for packaging. PMID- 10684285 TI - Replication of lengthened Moloney murine leukemia virus genomes is impaired at multiple stages. AB - It has been assumed that RNA packaging constraints limit the size of retroviral genomes. This notion of a retroviral "headful" was tested by examining the ability of Moloney murine leukemia virus genomes lengthened by 4, 8, or 11 kb to participate in a single replication cycle. Overall, replication of these lengthened genomes was 5- to 10-fold less efficient than that of native-length genomes. When RNA expression and virion formation, RNA packaging, and early stages of replication were compared, long genomes were found to complete each step less efficiently than did normal-length genomes. To test whether short RNAs might facilitate the packaging of lengthy RNAs by heterodimerization, some experiments involved coexpression of a short packageable RNA. However, enhancement of neither long vector RNA packaging nor long vector DNA synthesis was observed in the presence of the short RNA. Most of the proviruses templated by 12 and 16 kb vectors appeared to be full length. Most products of a 19. 2-kb vector contained deletions, but some integrated proviruses were around twice the native genome length. These results demonstrate that lengthy retroviral genomes can be packaged and that genome length is not strictly limited at any individual replication step. These observations also suggest that the lengthy read-through RNAs postulated to be intermediates in retroviral transduction can be packaged directly without further processing. PMID- 10684286 TI - Jembrana disease virus Tat can regulate human immunodeficiency virus (HIV) long terminal repeat-directed gene expression and can substitute for HIV Tat in viral replication. AB - Jembrana disease virus (JDV) is a bovine lentivirus genetically similar to bovine immunodeficiency virus; it causes an acute and sometimes fatal disease in infected animals. This virus carries a very potent Tat that can strongly activate not only its own long terminal repeat (LTR) but also the human immunodeficiency virus (HIV) LTR. In contrast, HIV Tat cannot reciprocally activate the JDV LTR (H. Chen, G. E. Wilcox, G. Kertayadnya, and C. Wood, J. Virol. 73:658-666, 1999). This indicates that in transactivation JDV Tat may utilize a mechanism similar to but not the same as that of the HIV Tat. To further study the similarity of JDV and HIV tat in transactivation, we first tested the responses of a series of HIV LTR mutants to the JDV Tat. Cross-transactivation of HIV LTR by JDV Tat was impaired by mutations that disrupted the HIV type 1 transactivation response element (TAR) RNA stem-loop structure. Our results demonstrated that JDV Tat, like HIV Tat, transactivated the HIV LTR at least partially in a TAR-dependent manner. However, the sequence in the loop region of TAR was not as critical for the function of JDV Tat as it was for HIV Tat. The competitive inhibition of Tat induced transactivation by the truncated JDV or HIV Tat, which consisted only of the activation domain, suggested that similar cellular factors were involved in both JDV and HIV Tat-induced transactivation. Based on the one-round transfection assay with HIV tat mutant proviruses, the cotransfected JDV tat plasmid can functionally complement the HIV tat defect. To further characterize the effect of JDV Tat on HIV, a stable chimeric HIV carrying the JDV tat gene was generated. This chimeric HIV replicated in a T-cell line, C8166, and in peripheral blood mononuclear cells, which suggested that JDV Tat can functionally substitute for HIV Tat. Further characterization of this chimeric virus will help to elucidate how JDV Tat functions and to explain the differences between HIV and JDV Tat transactivation. PMID- 10684287 TI - Role of hemagglutinin surface density in the initial stages of influenza virus fusion: lack of evidence for cooperativity. AB - Membrane fusion mediated by influenza virus hemagglutinin (HA) is believed to proceed via the cooperative action of multiple HA trimers. To determine the minimal number of HA trimers required to trigger fusion, and to assess the importance of cooperativity between these HA trimers, we have generated virosomes containing coreconstituted HAs derived from two strains of virus with different pH dependencies for fusion, X-47 (optimal fusion at pH 5.1; threshold at pH 5.6) and A/Shangdong (optimal fusion at pH 5.6; threshold at pH 6.0), and measured fusion of these virosomes with erythrocyte ghosts by a fluorescence lipid mixing assay. Virosomes with different X-47-to-A/Shangdong HA ratios, at a constant HA to-lipid ratio, showed comparable ghost-binding activities, and the low-pH induced conformational change of A/Shangdong HA did not affect the fusion activity of X-47 HA. The initial rate of fusion of these virosomes at pH 5.7 increased directly proportional to the surface density of A/Shangdong HA, and a single A/Shangdong trimer per virosome appeared to suffice to induce fusion. The reciprocal of the lag time before the onset of fusion was directly proportional to the surface density of fusion-competent HA. These results support the notion that there is no cooperativity between HA trimers during influenza virus fusion. PMID- 10684288 TI - Activation of lymphocyte signaling by the R1 protein of rhesus monkey rhadinovirus. AB - Rhesus monkey rhadinovirus (RRV) is a gamma-2 herpesvirus that exhibits a considerable degree of similarity to the human Kaposi's sarcoma-associated herpesvirus (KSHV). The R1 protein of RRV is distantly related to the K1 protein of KSHV, and R1, like K1, can contribute to cell growth transformation. In this study we analyzed the ability of the cytoplasmic tail of R1 to function as a signal transducer. The cytoplasmic domain of the R1 protein contains several tyrosine residues whose phosphorylation is induced in cells expressing Syk kinase. Expression of a CD8 chimera protein containing the extracellular and transmembrane domains of CD8 fused to the cytoplasmic domain of R1 mobilized intracellular calcium and induced cellular tyrosine phosphorylation in B cells upon stimulation with anti-CD8 antibody. None of the CD8-R1 cytoplasmic deletion mutants tested were able to mobilize intracellular calcium or to induce tyrosine phosphorylation to a significant extent upon addition of anti-CD8 antibody. Expression of wild-type R1 protein activated nuclear factor of activated T lymphocytes (NFAT) eightfold in B cells in the absence of antibody stimulation; expression of the CD8-R1C chimera strongly induced NFAT activity (60-fold) but only upon the addition of anti-CD8 antibody. We conclude that the cytoplasmic domain of R1 is capable of transducing signals that elicit B-lymphocyte activation events. The signal-inducing properties of R1 appear to be similar to those of K1 but differ in that the required sequences are distributed over a much longer stretch of the cytoplasmic domain (>150 amino acids). In addition, the induction of calcium mobilization was considerably longer in duration and stronger with R1 than with K1. PMID- 10684289 TI - Regulation of adenovirus membrane penetration by the cytoplasmic tail of integrin beta5. AB - Adenovirus (Ad) cell entry involves sequential interactions with host cell receptors that mediate attachment (CAR), internalization (alphavbeta3 and alphavbeta5), and penetration (alphavbeta5) of the endosomal membrane. These events allow the virus to deliver its genome to the nucleus. While integrins alphavbeta3 and alphavbeta5 both promote Ad internalization into cells, integrin alphavbeta5 selectively facilitates Ad-mediated membrane permeabilization and endosome rupture. In the experiments reported herein, we demonstrate that the intracellular domain of the integrin beta5 subunit specifically regulates Ad mediated membrane permeabilization and gene delivery. CS-1 melanoma cells expressing a truncated integrin beta5 or a chimeric (beta5-beta3) cytoplasmic tail (CT) supported normal levels of Ad endocytosis but had reduced Ad-mediated gene delivery and membrane permeabilization relative to cells expressing a wild type integrin beta5. Thin-section electron microscopy revealed that virion particles were capable of being endocytosed into cells expressing a truncated beta5CT, but they failed to escape cytoplasmic vesicles and translocate to the nucleus. Site-specific mutagenesis studies suggest that a C-terminal TVD motif in the beta5CT plays a major role in Ad membrane penetration. PMID- 10684290 TI - Comparative efficacy of recombinant modified vaccinia virus Ankara expressing simian immunodeficiency virus (SIV) Gag-Pol and/or Env in macaques challenged with pathogenic SIV. AB - Prior studies demonstrated that immunization of macaques with simian immunodeficiency virus (SIV) Gag-Pol and Env recombinants of the attenuated poxvirus modified vaccinia virus Ankara (MVA) provided protection from high levels of viremia and AIDS following challenge with a pathogenic strain of SIV (V. M. Hirsch et al., J. Virol. 70:3741-3752, 1996). This MVA-SIV recombinant expressed relatively low levels of the Gag-Pol portion of the vaccine. To optimize protection, second-generation recombinant MVAs that expressed high levels of either Gag-Pol (MVA-gag-pol) or Env (MVA-env), alone or in combination (MVA-gag-pol-env), were generated. A cohort of 24 macaques was immunized with recombinant or nonrecombinant MVA (four groups of six animals) and was challenged with 50 times the dose at which 50% of macaques are infected with uncloned pathogenic SIVsmE660. Although all animals became infected postchallenge, plasma viremia was significantly reduced in animals that received the MVA-SIV recombinant vaccines as compared with animals that received nonrecombinant MVA (P = 0.0011 by repeated-measures analysis of variance). The differences in the degree of virus suppression achieved by the three MVA-SIV vaccines were not significant. Most importantly, the reduction in levels of viremia resulted in a significant increase in median (P < 0.05 by Student's t test) and cumulative (P = 0.010 by log rank test) survival. These results suggest that recombinant MVA has considerable potential as a vaccine vector for human AIDS. PMID- 10684291 TI - A novel mechanism of resistance to mouse mammary tumor virus infection. AB - Exogenous mouse mammary tumor virus (MMTV) is carried from the gut of suckling pups to the mammary glands by lymphocytes and induces mammary gland tumors. MMTV induced tumor incidence in inbred mice of different strains ranges from 0 to as high as 100%. For example, mice of the C3H/HeN strain are highly susceptible, whereas mice of the I/LnJ strain are highly resistant. Of the different factors that together determine the susceptibility of mice to development of MMTV-induced mammary tumors, genetic elements play a major role, although very few genes that determine a susceptibility-resistance phenotype have been identified so far. Our data indicate that MMTV fails to infect mammary glands in I/LnJ mice foster nursed on viremic C3H/HeN females, even though the I/LnJ mammary tissue is not refractory to MMTV infection. Lymphocytes from fostered I/LnJ mice contained integrated MMTV proviruses and shed virus but failed to establish infection in the mammary glands of susceptible syngeneic (I x C3H.JK)F(1) females. Based on the susceptible-resistant phenotype distribution in N(2) females, both MMTV mammary gland infection and mammary gland tumor development in I/LnJ mice are controlled by a single locus. PMID- 10684292 TI - Functional characterization of the human immunodeficiency virus type 1 genome by genetic footprinting. AB - We present a detailed and quantitative analysis of the functional characteristics of the 1,000-nucleotide segment at the 5' end of the human immunodeficiency virus type 1 (HIV-1) RNA genome. This segment of the viral genome contains several important cis-acting sequences, including the TAR, polyadenylation, viral att site, minus-strand primer-binding site, and 5' splice donor sequences, as well as coding sequences for the matrix protein and the N-terminal half of the capsid protein. The genetic footprinting technique was used to determine quantitatively the abilities of 134 independent insertion mutations to (i) make stable viral RNA, (ii) assemble and release viral RNA-containing viral particles, and (iii) enter host cells, complete reverse transcription, enter the nuclei of host cells, and generate proviruses in the host genome by integration. All of the mutants were constructed and analyzed en masse, greatly decreasing the labor typically involved in mutagenesis studies. The results confirmed the presence of several previously known functional features in this region of the HIV genome and provided evidence for several novel features, including newly identified cis acting sequences that appeared to contribute to (i) the formation of stable viral transcripts, (ii) viral RNA packaging, and (iii) an early step in viral replication. The results also pointed to an unanticipated trans-acting role for the N-terminal portion of matrix in the formation of stable viral RNA transcripts. Finally, in contrast to previous reports, the results of this study suggested that detrimental mutations in the matrix and capsid proteins principally interfered with viral assembly. PMID- 10684293 TI - The kinetics of VP5 mRNA expression is not critical for viral replication in cultured cells. AB - We generated recombinant viruses in which the kinetics of expression of the leaky late VP5 mRNA was altered. We then analyzed the effect of such alterations on viral replication in cultured cells. The VP5 promoter and leader sequences from positions -36 to +20, containing the TATA box and an initiator element, were deleted and replaced with a strong early (dUTPase), an equal-strength leaky-late (VP16), or a strict-late (U(L)38) promoter. We found that recombinant viruses containing the dUTPase promoter inserted in the VP5 locus expressed VP5-encoding mRNA with early kinetics, while virus with the U(L)38 promoter inserted expressed such mRNA with strict-late kinetics. Further, in spite of differences in its functional architecture, the VP16 promoter fully substituted for the VP5 promoter. Western blot analysis demonstrated that the amounts of VP5 capsid protein produced by the recombinant viruses differed somewhat; however, on complementing C32 and noncomplementing Vero cells, such viruses replicated to titers equivalent to those of the rescued wild-type virus controls. Multistep virus growth in mouse embryo fibroblasts, rabbit skin cells, and Vero cells also demonstrated equivalent replication efficiencies for both recombinant and wild type viruses. Further, recombinant viruses did not show any impairment in their ability to replicate on serum-starved or quiescent human lung fibroblasts. We conclude that the kinetics of the essential VP5 mRNA expression is not critical for viral replication in cultured cells. PMID- 10684294 TI - Infectious entry pathway of adeno-associated virus and adeno-associated virus vectors. AB - We have investigated the infectious entry pathway of adeno-associated virus (AAV) and recombinant AAV vectors by assessing AAV-mediated gene transfer and by covalently conjugating fluorophores to AAV and monitoring entry by fluorescence microscopy. We examined AAV entry in HeLa cells and in HeLa cell lines which inducibly expressed a dominant interfering mutant of dynamin. The data demonstrate that AAV internalizes rapidly by standard receptor-mediated endocytosis from clathrin-coated pits (half-time <10 min). The lysosomotropic agents ammonium chloride and bafilomycin A(1) prevent AAV-mediated gene transfer when present during the first 30 min after the onset of endocytosis, indicating that AAV escapes from early endosomes yet requires an acidic environment for penetration into the cytosol. Following release from the endosome, AAV rapidly moves to the cell nucleus and accumulates perinuclearly beginning within 30 min after the onset of endocytosis. We present data indicating that escape of AAV from the endosome and trafficking of viral particles to the nucleus are unaffected by the presence of adenovirus, the primary helper virus for a productive AAV infection. Within 2 h, viral particles could be detected within the cell nucleus, suggesting that AAV enters the nucleus prior to uncoating. Interestingly, the majority of the intracellular virus particles remain in a stable perinuclear compartment even though gene expression from nuclear AAV genomes can be detected. This suggests that the process of nuclear entry is rate limiting or that AAV entry involves multiple pathways. Nevertheless, these data establish specific points in the AAV infectious entry process and have allowed the generation of a model for future expansion to specific cell types and AAV vector analysis in vivo. PMID- 10684295 TI - Lymphotoxin-alpha-deficient mice can clear a productive infection with murine gammaherpesvirus 68 but fail to develop splenomegaly or lymphocytosis. AB - Respiratory challenge with murine gammaherpesvirus 68 (MHV-68) leads to an acute productive infection of the lung and a persistent latent infection in B lymphocytes, epithelia, and macrophages. The virus also induces splenomegaly and an increase in the number of activated CD8 T cells in the circulation. Lymphotoxin- alpha-deficient (LTalpha(-/-)) mice have no lymph nodes and have disrupted splenic architecture. Surprisingly, in spite of the severe defect in secondary lymphoid tissue, LTalpha(-/-) mice could clear a productive MHV-68 infection, although with delayed kinetics compared to wild-type mice, and could control latent infection. Cytotoxic T-cell activity was comparable in the lungs and spleens of LTalpha(-/-) and wild-type mice. However, splenic gamma interferon responses were substantially reduced in LTalpha(-/-) mice. Furthermore, LTalpha( /-) mice failed to develop splenomegaly or lymphocytosis. Although germinal centers were absent, LTalpha(-/-) mice were able to class switch and showed significant virus-specific antibody titers. This work demonstrates that organized secondary lymphoid tissue is not an absolute requirement for the generation of immune responses to viral infections. PMID- 10684296 TI - Expression of two related viral early genes in Epstein-Barr virus-associated tumors. AB - The transcription of two early "leftwardly" expressed genes carrying repetitive sequences, IR2 and IR4, has been studied for Epstein-Barr virus-associated tumors, and for established B-cell lines, using sequence-specific probes generated for this purpose. Whereas the IR4 transcript was identified in every tumor and cell line assessed (except B95-8, with a deletion that removes the gene), expression of the IR2 gene was restricted to B lymphocytes. Though the promoters for both transcripts lie within homologous regions (D(L) and D(R)) in the viral genome, the IR2 promoter appears more tightly regulated. Detailed characterization of the IR4 transcript from a nasopharyngeal carcinoma tumor, C15, identifies a sequence variant of this gene that differs from those reported for B cells; in situ hybridization methods show transcription to be restricted to a subset of cells, with the strongest signals seen adjacent to host stroma. As with B cells in culture (Y. Gao, P. R. Smith, L. Karran, Q. L. Lu, and B. E. Griffin, J. Virol. 71:84-94, 1997), chemical induction enhanced transcriptional expression of the IR4 gene in the C15 tumor, although staining for both the IR4 antigen and that of the virus lytic switch, Zta, gave negative results. In a Burkitt's lymphoma biopsy specimen, however, both proteins were found expressed, notably in the same subset of cells. The data here and elsewhere (Gao et al., J. Virol., 1997) are consistent with a block to intracellular transport of the transcript(s) and suggest nuclear roles for it in tumors, possibly in RNA processing and viral lytic replication. Both roles could be fulfilled in the absence of translation. PMID- 10684297 TI - Identification of contact residues and definition of the CAR-binding site of adenovirus type 5 fiber protein. AB - The binding of adenovirus (Ad) fiber knob to its cellular receptor, the coxsackievirus and Ad receptor (CAR), promotes virus attachment to cells and is a major determinant of Ad tropism. Analysis of the kinetics of binding of Ad type 5 (Ad5) fiber knob to the soluble extracellular domains of CAR together (sCAR) and each immunoglobulin (Ig) domain (IgV and IgC2) independently by surface plasmon resonance demonstrated that the IgV domain is necessary and sufficient for binding, and no additional membrane components are required to confer high affinity binding to Ad5 fiber knob. Four Ad5 fiber knob mutations, Ser408Glu and Pro409Lys in the AB loop, Tyr477Ala in the DG loop, and Leu485Lys in beta strand F, effectively abolished high-affinity binding to CAR, while Ala406Lys and Arg412Asp in the AB loop and Arg481Glu in beta strand E significantly reduced the level of binding. Circular dichroism spectroscopy showed that these mutations do not disorder the secondary structure of the protein, implicating Ser408, Pro409, Tyr477, and Leu485 as contact residues, with Ala406, Arg412, and Arg481 being peripherally or indirectly involved in CAR binding. The critical residues have exposed side chains that form a patch on the surface, which thus defines the high affinity interface for CAR. Additional site-directed mutagenesis of Ad5 fiber knob suggests that the binding site does not extend to the adjacent subunit or toward the edge of the R sheet. These findings have implications for our understanding of the biology of Ad infection, the development of novel Ad vectors for targeted gene therapy, and the construction of peptide inhibitors of Ad infection. PMID- 10684298 TI - Herpesvirus mRNAs are sorted for export via Crm1-dependent and -independent pathways. AB - Cellular pre-mRNA splicing is inhibited by ICP27, a herpes simplex virus regulatory protein, resulting in the shutoff of host protein synthesis. Here we reveal that ICP27 also mediates the export of some virus RNAs via a Crm1 dependent pathway and present evidence that independent domains are required for these functions. Sorting of some viral mRNAs for nuclear export requires Crm1, while other virus mRNAs are exported via another pathway. PMID- 10684299 TI - Identification of a boundary domain adjacent to the potent human cytomegalovirus enhancer that represses transcription of the divergent UL127 promoter. AB - Transcriptional repression within a complex modular promoter may play a key role in determining the action of enhancer elements. In human cytomegalovirus, the major immediate-early promoter (MIEP) locus contains a highly potent and complex modular enhancer. Evidence is presented suggesting that sequences of the MIEP between nucleotide positions -556 and -673 function to prevent transcription activation by enhancer elements from the UL127 open reading frame divergent promoter. Transient transfection assays of reporter plasmids revealed repressor sequences located between nucleotides -556 and -638. The ability of these sequences to confer repression in the context of an infection was shown using recombinant viruses generated from a bacterial artificial chromosome containing an infectious human cytomegalovirus genome. In addition to repressor sequences between -556 and -638, infection experiments using recombinant virus mutants indicated that sequences between -638 and -673 also contribute to repression of the UL127 promoter. On the basis of in vitro transcription and transient transfection assays, we further show that interposed viral repressor sequences completely inhibit enhancer-mediated activation of not only the homologous but also heterologous promoters. These and other experiments suggest that repression involves an interaction of host-encoded regulatory factors with defined promoter sequences that have the property of proximally interfering with upstream enhancer elements in a chromatin-independent manner. Altogether, our findings establish the presence of a boundary domain that efficiently blocks enhancer-promoter interactions, thus explaining how the enhancer can work to selectively activate the MIEP. PMID- 10684300 TI - Hepatitis B virus X protein colocalizes to mitochondria with a human voltage dependent anion channel, HVDAC3, and alters its transmembrane potential. AB - Understanding the mechanism(s) of action of the hepatitis B virus (HBV)-encoded protein HBx is fundamental to elucidating the underlying mechanisms of chronic liver disease and hepatocellular carcinoma caused by HBV infection. In our continued attempts to identify cellular targets of HBx, we have previously reported the identification of a novel cellular protein with the aid of a yeast two-hybrid assay. This cellular gene was identified as a third member of the family of human genes that encode the voltage-dependent anion channel (HVDAC3). In the present study, physical interaction between HBx and HVDAC3 was established by standard in vitro and in vivo methods. Confocal laser microscopy of transfected cells with respective expression vectors colocalized HVDAC3 and HBx to mitochondria. This novel, heretofore unreported subcellular distribution of HBx in mitochondria implies a functional role of HBx in functions associated with mitochondria. Using a stable cationic fluorophore dye, CMXRos, we show that HBx expression in cultured human hepatoma cells leads to alteration of mitochondrial transmembrane potential. Such functional roles of HBx in affecting mitochondrial physiology have implications for HBV-induced liver injury and the development of hepatocellular carcinoma. PMID- 10684301 TI - Subcellular redistribution of Pit-2 P(i) transporter/amphotropic leukemia virus (A-MuLV) receptor in A-MuLV-infected NIH 3T3 fibroblasts: involvement in superinfection interference. AB - Amphotropic murine leukemia virus (A-MuLV) utilizes the Pit-2 sodium-dependent phosphate transporter as a cell surface receptor to infect mammalian cells. Previous studies established that infection of cells with A-MuLV resulted in the specific down-modulation of phosphate uptake mediated by Pit-2 and in resistance to superinfection with A-MuLV. To study the mechanisms underlying these phenomena, we constructed plasmids capable of efficiently expressing epsilon epitope- and green fluorescent protein (GFP)-tagged human Pit-2 proteins in mammalian cells. Overexpression of epsilon-epitope-tagged Pit-2 transporters in NIH 3T3 cells resulted in a marked increase in sodium-dependent P(i) uptake. This increase in P(i) uptake was specifically blocked by A-MuLV infection but not by infection with ecotropic MuLV (E-MuLV) (which utilizes a cationic amino acid transporter, not Pit-2, as a cell surface receptor). These data, together with the finding that the tagged Pit-2 transporters retained their A-MuLV receptor function, indicate that the insertion of epitope tags does not affect either retrovirus receptor or P(i) transporter function. The overexpressed epitope tagged transporters were detected in cell lysates, by Western blot analysis using both epsilon-epitope- and GFP-specific antibodies as well as with Pit-2 antiserum. Both the epitope- and GFP-tagged transporters showed almost exclusive plasma membrane localization when expressed in NIH 3T3 cells, as determined by laser scanning confocal microscopy. Importantly, when NIH 3T3 cells expressing these proteins were productively infected with A-MuLV, the tagged transporters and receptors were no longer detected in the plasma membrane but rather were localized to a punctate structure within the cytosolic compartment distinct from Golgi, endoplasmic reticulum, endosomes, lysosomes, and mitochondria. The intracellular Pit-2 pool colocalized with the virus in A-MuLV-infected cells. A similar redistribution of the tagged Pit-2 proteins was not observed following infection with E-MuLV, indicating that the redistribution of Pit-2 is not directly attributable to general effects associated with retroviral infection but rather is a specific consequence of A-MuLV-Pit-2 interactions. PMID- 10684302 TI - Role of the Gag matrix domain in targeting human immunodeficiency virus type 1 assembly. AB - Human immunodeficiency virus type 1 (HIV-1) particle formation and the subsequent initiation of protease-mediated maturation occur predominantly on the plasma membrane. However, the mechanism by which HIV-1 assembly is targeted specifically to the plasma membrane versus intracellular membranes is largely unknown. Previously, we observed that mutations between residues 84 and 88 of the matrix (MA) domain of HIV-1 Gag cause a retargeting of virus particle formation to an intracellular site. In this study, we demonstrate that the mutant virus assembly occurs in the Golgi or in post-Golgi vesicles. These particles undergo core condensation in a protease-dependent manner, indicating that virus maturation can occur not only on the plasma membrane but also in the Golgi or post-Golgi vesicles. The intracellular assembly of mutant particles is dependent on Gag myristylation but is not influenced by p6(Gag) or envelope glycoprotein expression. Previous characterization of viral revertants suggested a functional relationship between the highly basic domain of MA (amino acids 17 to 31) and residues 84 to 88. We now demonstrate that mutations in the highly basic domain also retarget virus particle formation to the Golgi or post-Golgi vesicles. Although the basic domain has been implicated in Gag membrane binding, no correlation was observed between the impact of mutations on membrane binding and Gag targeting, indicating that these two functions of MA are genetically separable. Plasma membrane targeting of Gag proteins with mutations in either the basic domain or between residues 84 and 88 was rescued by coexpression with wild type Gag; however, the two groups of MA mutants could not rescue each other. We propose that the highly basic domain of MA contains a major determinant of HIV-1 Gag plasma membrane targeting and that mutations between residues 84 and 88 disrupt plasma membrane targeting through an effect on the basic domain. PMID- 10684303 TI - Identification and analysis of the K5 gene of Kaposi's sarcoma-associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8 (HHV-8), belongs to the gammaherpesvirus subfamily and encodes approximately 80 open reading frames (ORFs). Among them are a few candidates for immediate-early genes (e.g., K5). We developed a monoclonal antibody (MAb), 328C7, against the K5 antigen. This MAb reacted with the K5 gene product by immunoscreening of a cDNA library from BCBL-1 cells, and this result was confirmed by transfection of the K5 ORF into Cos-7 cells. After induction of lytic infection by treatment with 12 O-tetradecanoylphorbol-13-acetate, MAb 328C7 reacted with an antigen in the cytoplasm of BCBL-1 and BC-3 cells as early as after 4 h of induction. Immunoelectron microscopy showed that the K5 antigen was situated mainly in the endoplasmic reticulum but was not present on the virion or in the nucleus. Northern blotting with a K5-specific probe revealed a single transcript of 1.2 kb, while Western blotting showed the antigen to be a 36-kDa polypeptide. The 5' and 3' ends were then determined by rapid amplification of cDNA, followed by sequencing of RACE products, and a splice was revealed upstream of the K5 ORF. K5 expression was unaffected by the respective DNA and protein synthesis inhibitors phosphonoformic acid and cycloheximide plus actinomycin D, confirming its immediate-early nature. Transient-transfection assays showed that the K5 promoter was transactivated by ORF 50 (KSHV Rta), a homolog of Epstein-Barr virus Rta, but the K5 gene product exhibited no transregulation of its own promoter or those of DNA polymerase and the human immunodeficiency virus type 1 long terminal repeat. This is the first such analysis of an immediate-early gene product; determination of its specific biological function requires further investigation. PMID- 10684304 TI - Bovine herpesvirus 1 U(L)3.5 interacts with bovine herpesvirus 1 alpha transinducing factor. AB - The bovine herpesvirus 1 (BHV-1) U(L)3.5 gene encodes a 126-amino-acid tegument protein. Homologs of U(L)3.5 are present in some alphaherpesviruses and have 20 to 30% overall amino acid homology that is concentrated in the N-terminal 50 amino acids. Mutant pseudorabies virus lacking U(L)3.5 is deficient in viral egress but can be complemented by BHV-1 U(L)3.5 (W. Fuchs, H. Granzow, and T. C. Mettenleiter, J. Virol. 71:8886-8892, 1997). The function of BHV-1 U(L)3.5 in BHV 1 replication is not known. To get a better understanding of its function, we sought to identify the proteins that interact with the BHV-1 U(L)3.5 protein. By using an in vitro pull-down assay and matrix-assisted laser desorption ionization mass spectrometry analysis, we identified BHV-1 alpha-transinducing factor (alphaBTIF) as a BHV-1 U(L)3. 5-interacting protein. The interaction was verified by coimmunoprecipitation from virus-infected cells using an antibody to either protein, by indirect immunofluorescence colocalization in both virus-infected and transfected cells, and by the binding of in vitro-translated proteins. In virus infected cells, U(L)3.5 and alphaBTIF colocalized in a Golgi-like subcellular compartment late in infection. In transfected cells, they colocalized in the nucleus. Deletion of 20 amino acids from the N terminus of U(L)3.5, but not 40 amino acids from the C terminus, abolished the U(L)3.5-alphaBTIF interaction both in vitro and in vivo. The interaction between U(L)3. 5 and alphaBTIF may be important for BHV-1 maturation and regulation of alphaBTIF transactivation activity. PMID- 10684305 TI - The intact retroviral Env glycoprotein of human foamy virus is a trimer. AB - Electron microscopy of negatively stained human foamy virus particles provides direct evidence for the trimeric nature of intact Env surface glycoproteins. Three-dimensional image reconstruction reveals that the Env trimer is a tapering spike 14 nm in length. The spikes were often arranged in hexagonal rings which shared adjacent Env trimers. PMID- 10684306 TI - Recombinant adeno-associated virus expressing human papillomavirus type 16 E7 peptide DNA fused with heat shock protein DNA as a potential vaccine for cervical cancer. AB - In this study, we explore a potential vaccine for human papillomavirus (HPV) induced tumors, using heat shock protein as an adjuvant, a peptide vaccine for safety, and adeno-associated virus (AAV) as a gene delivery vector. The tumor vaccine was devised by constructing a chimeric gene which contained HPV type 16 E7 cytotoxic T-lymphocyte (CTL) epitope DNA (M. C. Feltkamp, H. L. Smits, M. P. Vierboom, R. P. Minnaar, B. M. de Jongh, J. W. Drijfhout, J. ter Schegget, C. J. Melief, and W. M. Kast, Eur. J. Immunol. 23:2242-2249, 1993) fused with the heat shock protein gene as a tumor vaccine delivered via AAV. Our results demonstrate that this vaccine can eliminate tumor cells in syngeneic animals and induce CD4- and CD8-dependent CTL activity in vitro. Moreover, studies with knockout mice with distinct T-cell deficiencies confirm that CTL-induced tumor protection is CD4 and CD8 dependent. Taken together, the evidence indicates that this chimeric gene delivered by AAV has potential as a cervical cancer vaccine. PMID- 10684308 TI - Superantigen expression is driven by both mouse mammary tumor virus long terminal repeat-associated promoters in transgenic mice. AB - In addition to the usual retroviral promoter, the mouse mammary tumor virus (MMTV) long terminal repeat carries a second promoter located in the U3 region. Here we show that both of these promoters are independently able to give rise to superantigen activity in transgenic mice. The ability of multiple MMTV promoters to drive superantigen expression underscores its importance in the virus life cycle. PMID- 10684307 TI - Productive replication of human adenoviruses in mouse epidermal cells. AB - In contrast to most cells of mouse origin, cell lines derived from mouse epidermis are permissive for replication of human adenovirus type 5. The extent of epidermal cell differentiation correlated with the level of E1A expression and virus replication. Mouse epidermal cells may provide useful models for cancer therapy using replication-competent human adenoviruses. PMID- 10684309 TI - Interaction of yellow fever virus French neurotropic vaccine strain with monkey brain: characterization of monkey brain membrane receptor escape variants. AB - Binding of yellow fever virus wild-type strains Asibi and French viscerotropic virus and vaccine strains 17D and FNV to monkey brain and monkey liver cell membrane receptor preparations (MRPs) was investigated. Only FNV bound to monkey brain MRPs, while French viscerotropic virus, Asibi, and FNV all bound to monkey liver MRPs. Four monkey brain and two mouse brain MRP escape (MRP(R)) variants of FNV were selected at pH 7.6 and 6.0. Three monkey brain MRP(R) variants selected at pH 7.6 each had only one amino acid substitution in the envelope (E) protein in domain II (E-237, E-260, or E274) and were significantly attenuated in mice following intracerebral inoculation. Two of the variants were tested in monkeys and retained parental neurotropism following intracerebral inoculation at the dose tested. We speculate that this region of domain II is involved in binding of FNV E protein to monkey brain and is, in part, responsible for the enhanced neurotropism of FNV for monkeys. A monkey brain MRP(R) variant selected at pH 6.0 and two mouse brain MRP(R) variants selected at pH 7.6 were less attenuated in mice, and each had an amino acid substitution in the transmembrane region of the E protein (E-457 or E-458). PMID- 10684310 TI - Nef-induced major histocompatibility complex class I down-regulation is functionally dissociated from its virion incorporation, enhancement of viral infectivity, and CD4 down-regulation. AB - The N-terminal alpha-helix domain of the human immunodeficiency virus type 1 (HIV 1) Nef protein plays important roles in enhancement of viral infectivity, virion incorporation of Nef, and the down-regulation of major histocompatibility complex class I (MHC-I) expression on cell surfaces. In this study, we demonstrated that Met 20 in the alpha-helix domain was indispensable for the ability of Nef to modulate MHC-I expression but not for other events. We also showed that Met 20 was unnecessary for the down-regulation of CD4. These findings indicate that the region governing MHC-I down-regulation is proximate in the alpha-helix domain but is dissociated functionally from that determining enhancement of viral infectivity, virion incorporation of Nef, and CD4 down-regulation. PMID- 10684311 TI - Herpes simplex virus ICP27 induces cytoplasmic accumulation of unspliced polyadenylated alpha-globin pre-mRNA in infected HeLa cells. AB - Transcripts of most intron-bearing cellular genes must be processed by the splicing machinery in order to efficiently accumulate and gain access to the cytoplasm. However, we found that herpes simplex virus induces cytoplasmic accumulation of both spliced and unspliced polyadenylated alpha-globin RNAs in infected HeLa cells. Accumulation of the unspliced RNA required the immediate early protein ICP27, and ICP27 was sufficient (in combination with ICP4) to produce this effect in a transient-transfection assay. However, expression of ICP27 did not markedly alter the levels of fully spliced alpha-globin transcripts in infected cells. These data demonstrate that the previously documented effects of ICP27 on the cellular splicing apparatus do not greatly inhibit splicing of alpha-globin RNA and argue that ICP27 induces a splicing-independent pathway for alpha-globin RNA accumulation and nuclear export. PMID- 10684312 TI - Enhancement of immunoglobulin G2a and cytotoxic T-lymphocyte responses by a booster immunization with recombinant hepatitis C virus E2 protein in E2 DNA primed mice. AB - The induction of strong cytotoxic T-lymphocyte (CTL) and humoral responses appear to be essential for the elimination of persistently infecting viruses, such as hepatitis C virus (HCV). Here, we tested several vaccine regimens and demonstrate that a combined vaccine regimen, consisting of HCV E2 DNA priming and boosting with recombinant E2 protein, induces the strongest immune responses to HCV E2 protein. This combined vaccine regimen augments E2-specific immunoglobulin G2a (IgG2a) and CD8(+) CTL responses to a greater extent than immunizations with recombinant E2 protein and E2 DNA alone, respectively. In addition, the data showed that a protein boost following one DNA priming was also effective, but much less so than those following two DNA primings. These data indicate that sufficient DNA priming is essential for the enhancement of DNA encoded antigen specific immunity by a booster immunization with recombinant E2 protein. Furthermore, the enhanced CD8(+) CTL and IgG2a responses induced by our combined vaccine regimens are closely associated with the protection of BALB/c mice from challenge with modified CT26 tumor cells expressing HCV E2 protein. Together, our results provide important implications for vaccine development for many pathogens, including HCV, which require strong antibody and CTL responses. PMID- 10684313 TI - A 13-amino-acid Pit1-specific loop 4 sequence confers feline leukemia virus subgroup B receptor function upon Pit2. AB - Feline leukemia virus subgroup B (FeLV-B) and gibbon ape leukemia virus (GALV) utilize the human protein Pit1 but not the related protein, Pit2, as receptor. A stretch of 9 amino acids, named region A, was identified in the putative fourth extracellular loop of Pit1 (residues 550 through 558) as critical for FeLV-B and GALV receptor function. However, the presence of Pit1 region A did not confer receptor function for FeLV-B upon Pit2, while it did so for GALV. We have here shown that the presence of two Pit1-specific loop 4 residues (tyrosine 546 and valine 548) in addition to Pit1 region A is sufficient to make Pit2 an efficient FeLV-B receptor; that is, a stretch of 13 amino acids encompassing all loop 4 amino acid differences between Pit1 and Pit2 comprises a C-terminal determinant for FeLV-B receptor function. Thus, the same limited receptor region is sufficient to confer receptor function for both viruses upon Pit2. PMID- 10684314 TI - The SU and TM envelope protein subunits of bovine leukemia virus are linked by disulfide bonds, both in cells and in virions. AB - After the polyprotein precursor of retroviral envelope proteins is proteolytically cleaved, the surface (SU) and transmembrane (TM) subunits remain associated with each other by noncovalent interactions or by disulfide bonds. Disulfide linkages confer a relatively stable association between the SU and TM envelope protein subunits of Rous sarcoma virus and murine leukemia virus. In contrast, the noncovalent association between SU and TM of human immunodeficiency virus leads to significant shedding of SU from the surface of infected cells. The SU and TM proteins of bovine leukemia virus (BLV) initially were reported to be disulfide linked but later were concluded not to be, since TM is often lost during purification of SU protein. Here, we show that SU and TM of BLV do, indeed, associate through disulfide bonds, whether the envelope proteins are overexpressed in transfected cells, are produced in virus-infected cells, or are present in newly produced virions. PMID- 10684315 TI - Mutational analysis of adeno-associated virus type 2 Rep68 protein endonuclease activity on partially single-stranded substrates. AB - The endonuclease activity of the Rep68 and Rep78 proteins (Rep68/78) of adeno associated virus type 2 (AAV) cuts at the terminal resolution site (trs) within the hairpin structure formed by the AAV inverted terminal repeats. Recent studies suggest that a DNA unwinding function of Rep68/78 may be required for endonuclease activity. We demonstrate that several mutant proteins which are endonuclease negative on a fully duplex hairpin substrate are endonuclease positive on a partially single-stranded hairpin substrate. Truncation analysis revealed that the endonuclease function is contained within the first 200 amino acids of Rep68/78. This endonucleolytic cleavage is believed to involve the covalent attachment of Rep68/78 to the trs via a phosphate-tyrosine linkage. A previous report (S. L. Walker, R. S. Wonderling, and R. A. Owens, J. Virol. 71:2722-2730, 1997) suggested that tyrosine 152 was part of the active site. We individually mutated each tyrosine within the first 200 amino acids of the Rep68 moiety of a maltose binding protein-Rep68/78 fusion protein to phenylalanine. Only mutation of tyrosine 156 resulted in a protein incapable of covalent attachment to a partially single-stranded hairpin substrate, suggesting that tyrosine 156 is part of the endonuclease active site. PMID- 10684316 TI - Functional reconstitution of thymopoiesis after human immunodeficiency virus infection. AB - We have utilized combination antiretroviral therapy following human immunodeficiency virus type 1-induced human CD4(+) thymocyte depletion in the SCID-hu mouse to examine the immune competence of reconstituting thymocytes which appear following administration of combination therapy. These cells express a normal distribution of T-cell receptor variable gene families and are responsive to costimulatory signals. These results suggest that normal thymic function may be restored following antiretroviral treatment. PMID- 10684317 TI - Characterization of the R572T point mutant of a putative cleavage site in human foamy virus Env. AB - A putative cleavage site of the human foamy virus (HFV) envelope glycoprotein (Env) was altered. Transient env expression revealed that the R572T mutant Env was normally expressed and modified by asparagine-linked oligosaccharide chains. However, this single-amino-acid substitution was sufficient to abolish all detectable cleavage of the gp130 precursor polyprotein. Cell surface biotinylation demonstrated that the uncleaved mutant gp130 was transported to the plasma membrane. The uncleaved mutant protein was incapable of syncytium formation. Glycoprotein-driven virion budding, a unique aspect of HFV assembly, occurred despite the absence of Env cleavage. We then substituted the R572T mutant env into a replication-competent HFV molecular clone. Transfection of the mutant viral DNA into BHK-21 cells followed by viral titration with the FAB (foamy virus-activated beta-galactosidase expression) assay revealed that proteolysis of the HFV Env was essential for viral infectivity. Wild-type HFV Env partially complemented the defective virus phenotype. Taken together, these experimental results established the location of the HFV Env proteolytic site; the effects of cleavage on Env transport, processing, and function; and the importance of Env proteolysis for virus maturation and infectivity. PMID- 10684318 TI - Hepadnavirus infection in captive gibbons. AB - The recent isolation of a nonhuman primate hepadnavirus from woolly monkeys prompted an examination of other primates for potentially new hepadnaviruses. A serological analysis of 30 captive gibbons revealed that 47% were positive for at least one marker of ongoing or previous infection with a hepatitis B virus (HBV). The amino acid sequences of the core and surface genes of human and gibbon virus isolates were very similar. Phylogenetic analysis indicated that the gibbon isolates lie within the human HBV family, indicating that these HBV isolates most likely stem from infection of gibbons from a human source. PMID- 10684319 TI - Recombinant modified vaccinia virus ankara expressing the surface gp120 of simian immunodeficiency virus (SIV) primes for a rapid neutralizing antibody response to SIV infection in macaques. AB - Neutralizing antibodies were assessed before and after intravenous challenge with pathogenic SIVsmE660 in rhesus macaques that had been immunized with recombinant modified vaccinia virus Ankara expressing one or more simian immunodeficiency virus gene products (MVA-SIV). Animals received either MVA-gag-pol, MVA-env, MVA gag-pol-env, or nonrecombinant MVA. Although no animals were completely protected from infection with SIV, animals immunized with recombinant MVA-SIV vaccines had lower virus loads and prolonged survival relative to control animals that received nonrecombinant MVA (I. Ourmanov et al., J. Virol. 74:2740-2751, 2000). Titers of neutralizing antibodies measured with the vaccine strain SIVsmH-4 were low in the MVA-env and MVA-gag-pol-env groups of animals and were undetectable in the MVA-gag-pol and nonrecombinant MVA groups of animals on the day of challenge (4 weeks after final immunization). Titers of SIVsmH-4-neutralizing antibodies remained unchanged 1 week later but increased approximately 100-fold 2 weeks postchallenge in the MVA-env and MVA-gag-pol-env groups while the titers remained low or undetectable in the MVA-gag-pol and nonrecombinant MVA groups. This anamnestic neutralizing antibody response was also detected with T-cell-line adapted stocks of SIVmac251 and SIV/DeltaB670 but not with SIVmac239, as this latter virus resisted neutralization. Most animals in each group had high titers of SIVsmH-4-neutralizing antibodies 8 weeks postchallenge. Titers of neutralizing antibodies were low or undetectable until about 12 weeks of infection in all groups of animals and showed little or no evidence of an anamnestic response when measured with SIVsmE660. The results indicate that recombinant MVA is a promising vector to use to prime for an anamnestic neutralizing antibody response following infection with primate lentiviruses that cause AIDS. However, the Env component of the present vaccine needs improvement in order to target a broad spectrum of viral variants, including those that resemble primary isolates. PMID- 10684320 TI - Pitfalls in the interpretation of common biochemical tests. AB - This review considers some of the more common problems in the interpretation of the results of biochemical tests and, where possible, highlights ways in which errors can be identified or avoided. PMID- 10684321 TI - Amiodarone-induced thyroid disorders: a clinical review. AB - Although amiodarone is regarded as a highly effective anti-arrhythmic agent, its use may lead to alterations in thyroid gland function and/or thyroid hormone metabolism, partly because of its rich iodine content. Patients treated with amiodarone may manifest altered thyroid hormone profile without thyroid dysfunction, or they may present with clinically significant amiodarone-induced hypothyroidism or amiodarone-induced thyrotoxicosis. The former results from the inability of the thyroid to escape from the Wolff-Chaikoff effect. It prevails in areas with high dietary iodine intake, and it is readily managed by discontinuation of amiodarone or thyroid hormone replacement. Amiodarone-induced thyrotoxicosis occurs more frequently in areas with low iodine intake; it may arise from iodine-induced excessive thyroid hormone synthesis (type I) or destructive thyroiditis with release of preformed hormones (type II). Type I should be treated with thionamides alone or in combination with potassium perchlorate, whereas type II benefits from treatment with glucocorticoids. Surgery may be a feasible option for patients who require long-term amiodarone treatment. PMID- 10684322 TI - Helicobacter pylori. AB - Helicobacter pylori infection is a major cause of peptic ulcer disease, and its detection and eradication are now an important part of gastroenterology. Effective regimes are available which will eliminate the organism in about 90% of cases in developed countries. PMID- 10684323 TI - Carbohydrate-deficient glycoprotein syndromes. AB - Four types of carbohydrate-deficient glycoprotein syndrome have been described, and the cause of two of them has been found. The symptoms and signs of these syndromes are described, with variations that occur at different ages. The commonest is type Ia with an autosomal recessive form of inheritance, and the gene responsible has been mapped to 16p. The typical pathology is atrophy of the cerebellum and brainstem, sometimes also involving the cortex, although both the pathology and the biochemical deficiencies vary between different types of syndrome. The diagnosis depends firstly on recognising the clinical features, including the presence of complications such as thyroid disorders. Then biochemical tests can be carried out, especially chromatographic carbohydrate deficient transferrin assay and isoelectric focusing of serum transferrin. The prognosis depends on the complications, renal, hepatic, and cardiac, but affected children will be severely handicapped. Therefore treatment consists mainly of coping with the complications, and supporting the child and the family. Oral infusion of mannose can be effective in type Ib disease. PMID- 10684324 TI - Evaluation of patients admitted with hypoglycaemia to a teaching hospital in Central Anatolia. AB - Hypoglycaemia is one of the most common endocrine emergencies in practice. We analysed retrospectively the incidence and causes of hypoglycaemia in patients admitted to Erciyes University Medical School in Turkey between January 1991 and June 1998 because of hypoglycaemia. Charts were reviewed according to multiple variables including age, sex, blood glucose levels, renal and liver functions, diagnoses, symptoms, drugs, complications, sequelae, and survival status. During this period, 13 500 patients were hospitalised and hypoglycaemia was reported in 126 (0.9%) patients. The most common causes were diabetic treatments in 54 cases (42%), endocrine deficiencies in 25 cases (20%), and malignancy in 15 cases (12%), respectively. The leading endocrine deficiency was panhypopituitarism. Sheehan's syndrome was the most common cause of panhypopituitarism (44%). No underlying cause was diagnosed in seven cases. Nine patients died (7%) and neurological sequelae were observed in one patient with diabetes mellitus. We conclude that hypoglycaemia accounts for about 1% of hospital admissions. Although the hypoglycaemia could be attributed to hypoglycaemic agents in the treatment of diabetes mellitus in the majority of cases, Sheehan's syndrome was also found to be an important cause of hypoglycaemia in our hospital. PMID- 10684325 TI - Prevalence of mental illness in a rehabilitation unit for older adults. AB - The prevalence of psychiatric disorders was studied in 78 elderly people in a rehabilitation unit for older adults. The patients were assessed using the Evans Liverpool depression rating scale, Hospital Anxiety and Depression scale and Mini Mental State Examination. Twenty-eight (35.9%) patients were found to be depressed, 15 of these also had raised anxiety. Thirty-one (41.0%) patients had significant cognitive impairment and 14 of these had associated depression. Only 33 (42%) had no evidence of either cognitive impairment or mood disorder. On discharge, 20 (25.6%) patients were on antidepressant treatment but only 50% of those had this diagnosis recorded on the discharge summary. Our results showed higher prevalence of depression in this situation compared with the reported prevalence of 20-30% in the acute hospital setting. We recommend that all patients undergoing rehabilitation should be routinely screened for depression as it is common and treatment will improve the overall outcome. PMID- 10684326 TI - Day-case percutaneous endoscopic gastrostomy: a viable proposition? AB - The aim of our study was to evaluate the success rate, complications, and long term outcomes following day-case percutaneous endoscopic gastrostomy (PEG). This retrospective study was carried out in a 650-bed District General hospital in Northamptonshire, UK. Thirty-six patients, aged 28-90 years, were included in the study, 21 males (58%) and 15 females (42%). Indications for PEG insertion included head and neck cancer, dysphagia as a result of primary disease, and AIDS related malnutrition. Data were collected from the medical and dietetic records. The PEG procedure was successful in 33 patients (92%). In 32 cases (97%) the patient was discharged home. Twenty five of the patients (76%) suffered no complications whilst seven (21%) suffered complications within a month of the procedure. No patient required further surgical intervention. Five patients (15%) died of their primary disease within a month of the procedure. Patients had had their PEG tubes in situ for up to 2.5 years at the end of data collection. We conclude that PEG can be performed as a day-case procedure in stable patients with no increase in complication rate, morbidity, or mortality. PMID- 10684327 TI - Recurrence of adrenal aldosterone-producing adenoma. AB - Conn's syndrome (adrenal aldosterone-producing adenoma) and bilateral adrenal hyperplasia are the most common causes of primary aldosteronism. The treatment of choice for patients with aldosterone-producing adenoma is unilateral total adrenalectomy. Recurrence after adequate surgery is exceptional. We present a patient with recurrence of an aldosterone-producing adenoma in the right adrenal gland 9 years after adenomectomy of a aldosterone-producing adenoma in the same adrenal gland. We conclude that adenomectomy is not an adequate therapy for patients with adrenal aldosterone-producing adenoma. PMID- 10684328 TI - Can myasthenia gravis be diagnosed with the 'ice pack test'? A cautionary note. AB - The ice pack test may be helpful in establishing that ptosis is due to ocular myasthenia gravis, since cold improves neuromuscular transmission. However, the role of the test in determining whether diplopia is of myasthenic origin has yet to be established. PMID- 10684329 TI - Syncope after effort. AB - A 29-year-old man developed recurrent syncope following exertion. Cardiac investigations revealed no evidence of structural heart disease, but during exercise testing, in the recovery phase, he sustained a bradycardia and then asystole for a prolonged period. Before cardiac massage could be instituted a tonic-clonic fit occurred, and this initiated a return to sinus rhythm. His symptoms were abolished following the implantation of a dual-chamber pacemaker. PMID- 10684330 TI - Brittle Addison's disease: a new variation on a familiar theme. AB - Unstable and unpredictable disease control in diabetes or asthma, with frequent hospitalisations, is frequently referred to as 'brittle'. We describe two cases of Addison's disease with recurrent hospitalisations in hypo-adrenal crises. Both patients had significant psychosocial disruption, and failure to take hydrocortisone replacement therapy was admitted in one and biochemically proven in the other. We propose that 'brittle' Addison's disease in these cases was due to poor treatment compliance related to psychosocial factors. These features have particular similarities with the syndrome of brittle diabetes. PMID- 10684331 TI - Chest pain, enzymes and hypothyroidism. AB - Hypothyroidism is a common disorder and when presenting with classical symptoms and signs is easy to recognise. However, hypothyroidism may present in a manner suggestive of an acute myocardial infarction with an elevated creatine kinase and electrocardiographic abnormalities. We report a case of severe hypothyroidism presenting as a cardiac event whose symptoms and signs dispersed following treatment with thyroxine. PMID- 10684332 TI - Rapidly progressing tetraparesis in a young male patient with pyrexia. PMID- 10684333 TI - Hyperthyroidism in an elderly patient. PMID- 10684335 TI - Isosexual precocity: uncommon presentation of a common disorder. PMID- 10684334 TI - A case of intestinal obstruction. PMID- 10684336 TI - Calf hypertrophy following paralytic poliomyelitis. PMID- 10684337 TI - A healthy patient who suddenly developed a foot-drop. PMID- 10684338 TI - An unusual cause of bowel obstruction. PMID- 10684339 TI - Disseminated intravascular coagulation and vasculitis during propylthiouracil therapy. PMID- 10684341 TI - A colour handbook of dermatology PMID- 10684340 TI - Virtual endoscopy of the upper airway--a diagnostic tool. PMID- 10684342 TI - Sigmundoscopy. medical-psychiatric consultation-liaison. The bases PMID- 10684343 TI - Guidelines in clinical practice PMID- 10684344 TI - Molecular evolution of the GATA family of transcription factors: conservation within the DNA-binding domain. AB - The GATA-binding transcription factors comprise a protein family whose members contain either one or two highly conserved zinc finger DNA-binding domains. Members of this group have been identified in organisms ranging from cellular slime mold to vertebrates, including plants, fungi, nematodes, insects, and echinoderms. While much work has been done describing the expression patterns, functional aspects, and target genes for many of these proteins, an evolutionary analysis of the entire family has been lacking. Herein we show that only the C terminal zinc finger (Cf) and basic domain, which together constitute the GATA binding domain, are conserved throughout this protein family. Phylogenetic analyses of amino acid sequences demonstrate distinct evolutionary pathways. Analysis of GATA factors isolated from vertebrates suggests that the six distinct vertebrate GATAs are descended from a common ancestral sequence, while those isolated from nonvertebrates (with the exception of the fungal AREA orthologues and Arabidopsis paralogues) appear to be related only within the DNA-binding domain and otherwise provide little insight into their evolutionary history. These results suggest multiple modes of evolution, including gene duplication and modular evolution of GATA factors based upon inclusion of a class IV zinc finger motif. As such, GATA transcription factors represent a group of proteins related solely by their homologous DNA-binding domains. Further analysis of this domain examines the degree of conservation at each amino acid site using the Boltzmann entropy measure, thereby identifying residues critical to preservation of structure and function. Finally, we construct a predictive motif that can accurately identify potential GATA proteins. PMID- 10684345 TI - The presence of GSI-like genes in higher plants: support for the paralogous evolution of GSI and GSII genes. AB - Glutamine synthetase type I (GSI) genes have previously been described only in prokaryotes except that the fungus Emericella nidulans contains a gene (fluG) which encodes a protein with a large N-terminal domain linked to a C-terminal GSI like domain. Eukaryotes generally contain the type II (GSII) genes which have been shown to occur also in some prokaryotes. The question of whether GSI and GSII genes are orthologues or paralogues remains a point of controversy. In this article we show that GSI-like genes are widespread in higher plants and have characterized one of the genes from the legume Medicago truncatula. This gene is part of a small gene family and is expressed in many organs of the plant. It encodes a protein similar in size and with between 36 and 46% amino acid sequence similarity to prokaryotic GS proteins used in the analyses, whereas it is larger and with less than 25% similarity to GSII proteins, including those from the same plant species. Phylogenetic analyses suggest that this protein is most similar to putative proteins encoded by expressed sequence tags of other higher plant species (including dicots and a monocot) and forms a cluster with FluG as the most divergent of the GSI sequences. The discovery of GSI-like genes in higher plants supports the paralogous evolution of GSI and GSII genes, which has implications for the use of GS in molecular studies on evolution. PMID- 10684346 TI - Disparate evolution of paralogous introns in the Xdh gene of Drosophila. AB - Drosophila nuclear introns are commonly assumed to change according to a single rate of substitution, yet little is known about the evolution of these non-coding sequences. The hypothesis of a uniform substitution rate for introns seems to be at odds with recent findings that the nucleotide composition of introns varies at a scale unknown before, and that their base content variation is correlated with that of the adjacent exons. However, no direct attempt at comparing substitution rates in introns seems to have been addressed so far. We have studied the rate of nucleotide substitution over a region of the Xdh gene containing two adjacent short, constitutively spliced introns, in several species of Drosophila and related genera. The two introns differ significantly in base composition and substitution rate, with one intron evolving at least twice as fast as the other. In addition, the substitution pattern of the introns is positively associated with that of the surrounding coding regions, evidencing that the molecular evolution of these introns is impacted by the region in which they are embedded. The observed differences cannot be attributed to selection acting differently at the level of the secondary structure of the pre-mRNA. Rather, they are better accounted for by locally heterogeneous patterns of mutation. PMID- 10684347 TI - The codon-degeneracy model of molecular evolution. AB - Mitochondrial genetic codons can be categorized by four patterns of nucleotide site degeneracy based on varying combinations of twofold- or nondegenerate sites at first codon positions and twofold- or fourfold-degenerate sites at third codon positions. Herein, a model of molecular evolution is introduced that uses these patterns to calculate expected substitution frequencies for each codon position and substitution type relative to overall number of synonymous or nonsynonymous substitutions. Regions of the pocket gopher cytochrome oxidase subunit I (COI) and cytochrome b (cyt-b) genes are analyzed using this model. Chi-square distributions are used to produce relative goodness-of-fit (GF) scores for measuring the difference between substitution frequencies predicted by the codon degeneracy model (CDM), and frequencies inferred using a well-supported phylogenetic tree of closely related species. The GF scores for expected and observed synonymous (GF(syn) = 0.429, p = 0.807) and nonsynonymous (GF(ns) = 2.309, p = 0.679) substitution frequencies resulted in a failure to reject the CDM as a null hypothesis for the molecular evolution of COI and cyt-b in pocket gophers. Alternative tree topologies and calculations of transition bias for these data result in higher GF scores. PMID- 10684348 TI - Neutral and nonneutral mitochondrial genetic variation in deep-sea clams from the family vesicomyidae. AB - Nucleotide sequences at two mitochondrial genes from 57 individuals representing eight species of deep-sea clams (Vesicomyidae) were examined for variation consistent with the neutral model of molecular evolution. One gene, cytochrome oxidase subunit I (COI), deviated from the expectations of neutrality by containing an excess of intraspecific nonsynonymous polymorphism. Additionally, one species, Calyptogena kilmeri, showed a significant excess of rare polymorphism specifically at the COI locus. In contrast, a second mitochondrial gene, the large-subunit 16S ribosomal RNA gene (16S), showed little deviation from neutrality either between or within species. Together, COI and 16S show no deviation from neutral expectations by the HKA test, produce congruent phylogenetic relationships between species, and show correlated numbers of fixed differences between species and polymorphism within species. These patterns of both neutral and nonneutral evolution within the mitochondrial genome are most consistent with a model where intraspecific nonsynonymous polymorphism at COI is near neutrality. In addition to examining the forces of molecular evolution, we extend hypotheses about interspecific relationships within this family for geographical locations previously unexamined by molecular methods including habitats near the Middle Atlantic, the Aleutian Trench, and Costa Rica. PMID- 10684349 TI - Concerted evolution of a highly repetitive DNA family in eptatretidae (Cyclostomata, agnatha) implies specifically differential homogenization and amplification events in their germ cells. AB - In eight hagfish species, it is known that chromosome elimination occurs during early embryogenesis, and some highly repetitive DNA families, restricted to germ cells, have been isolated. One of these families, "EEEo2," has been isolated as DNA fragments by restriction enzyme analyses from Eptatretus okinoseanus and E. cirrhatus. In this study, EEEo2 sequences were isolated from germline DNA in E. burgeri, Paramyxine sheni, and P. atami using PCR methods. Sequence analysis revealed that these sequences are intraspecifically homogeneous, except in E. burgeri, and are interspecifically conserved with heterogeneity. The intraspecific sequence variability tends to decrease as the copy number increases. These results indicate that EEEo2 has evolved in a concerted manner. Moreover, an ancestral repeating motif consisting of triplicate subrepeats was deduced. These results suggest that EEEo2 arose as an initial amplification of this subrepeat and has evolved by saltatory replication. Phylogenetic analyses suggested the possibility that EEEo2 in E. okinoseanus and E. cirrhatus has been subjected to strong homogenizing forces for concerted evolution, whereas the force is weak in E. burgeri. In addition, EEEo2 in P. sheni and P. atami appear to have been incompletely subjected to these forces. Chromosomal in situ hybridization experiments revealed that EEEo2 sequences were located along almost their entire length of several heterochromatic chromosomes that are restricted to germ cells. These chromosomes are disposed to form a secondary association during the first meiotic metaphases, except in P. sheni. This chromosomal distribution may promote a concerted mode of sequence evolution in both nonhomologous chromosomes and homologous chromosomes and reflect the differential driving forces between species. PMID- 10684350 TI - Gene conversions may obscure actin gene family relationships. AB - Phylogenetic hypotheses of muscle actin evolution are significantly different when a sea urchin is used as a representative echinoderm than when a sea star is used. While sea urchin muscle actins support an echinoderm-chordate sister relationship, sea star sequences suggest that echinoderm muscle actins are convergent with chordate muscle actins. Our results suggest that gene conversion in the sea star muscle actin may be responsible for these discordant results. PMID- 10684351 TI - Do pheromone binding proteins converge in amino acid sequence when pheromones converge? AB - Convergence in amino acid sequences between proteins can be strong evidence for selection. Here, I look for evidence of convergence in the amino acid sequences of pheromone binding protein (PBP) in response to convergence in pheromones. PBPs are involved in sex pheromone reception by the antennae of male moths. In this role PBPs may selectively bind pheromone components and experience convergent selection in response to convergence in pheromone components. However, examination of the PBPs of the taxa that have converged upon the use of (E)- or (Z)-11-tetradecenyl acetate as their major pheromone component reveals little evidence for convergence in the PBPs identified from these taxa. A few sites show a pattern consistent with convergence or parallelism; however, it cannot be ruled out that these sites share the ancestral state. Two of these sites fall within the proposed binding region of PBPs. These results suggest that PBPs either have not converged in sequence or have converged at very few sites in response to convergence on the same pheromone component. PMID- 10684352 TI - Codon usage in plastid genes is correlated with context, position within the gene, and amino acid content. AB - Highly expressed plastid genes display codon adaptation, which is defined as a bias toward a set of codons which are complementary to abundant tRNAs. This type of adaptation is similar to what is observed in highly expressed Escherichia coli genes and is probably the result of selection to increase translation efficiency. In the current work, the codon adaptation of plastid genes is studied with regard to three specific features that have been observed in E. coli and which may influence translation efficiency. These features are (1) a relatively low codon adaptation at the 5' end of highly expressed genes, (2) an influence of neighboring codons on codon usage at a particular site (codon context), and (3) a correlation between the level of codon adaptation of a gene and its amino acid content. All three features are found in plastid genes. First, highly expressed plastid genes have a noticeable decrease in codon adaptation over the first 10-20 codons. Second, for the twofold degenerate NNY codon groups, highly expressed genes have an overall bias toward the NNC codon, but this is not observed when the 3' neighboring base is a G. At these sites highly expressed genes are biased toward NNT instead of NNC. Third, plastid genes that have higher codon adaptations also tend to have an increased usage of amino acids with a high G + C content at the first two codon positions and GNN codons in particular. The correlation between codon adaptation and amino acid content exists separately for both cytosolic and membrane proteins and is not related to any obvious functional property. It is suggested that at certain sites selection discriminates between nonsynonymous codons based on translational, not functional, differences, with the result that the amino acid sequence of highly expressed proteins is partially influenced by selection for increased translation efficiency. PMID- 10684353 TI - Lack of evidence for cospeciation between retroelements and their hosts. AB - Some literature is available on cospeciation and on reconstructing the phylogenetic relationships of retroelements, but relatively little consideration has been given to whether there is cospeciation between retroelements and their hosts. Here we address this problem in detail. We conclude that there is no significant evidence for cospeciation between retroelements and their hosts. This conclusion was reached by noting that the branching order of the two phylogenies was no more similar than would be expected by chance. PMID- 10684354 TI - A probable mixed-function supraoperon in pseudomonas exhibits gene organization features of both intergenomic conservation and gene shuffling PMID- 10684355 TI - Pharmacokinetics and tolerance of mycophenolate mofetil in renal transplant children. AB - Mycophenolate mofetil (MMF) is a prodrug that is hydrolyzed to the active immunosuppressant mycophenolic acid (MPA). The drug is now widely prescribed for adult renal transplant recipients and its use has been extended to pediatric patients, although pharmacological data in this age group are limited. Nine pediatric renal transplant recipients received MMF with corticosteroids and either cyclosporine or tacrolimus a median of 55 months (range 7.5-124 months) months after transplantation. The pharmacokinetic parameters of MPA and MPA glucuronide (MPAG) were determined at steady state by high-performance liquid chromatography after administration of MMF at the oral dose of 494+/-142 mg/m(2) twice daily. MPA was rapidly absorbed, with a peak concentration at 1.4 h. The mean plasma concentration of MPA at steady state was 4.7+/-1.3 microg/ml. The areas under the plasma concentration-time curves (AUCs) over 12 h (between two administrations) were 57.0+/-15.3 microg.h/ml for MPA and 1,515+/-722 microg.h/ml for MPAG, and the apparent oral clearance was 11.7+/-7.0 and 0.5+/-0.4 l/h for MPA and MPAG, respectively. Assuming that the pharmacokinetics of MPA was dose dependent, the mean concentration at steady state and the AUC for MPA were calculated for the recommended dosage schedule of 600 mg/m(2) every 12 h and were 6.3+/-2.7 microg/ml and 75.2+/-32.9 microg.h/ml, respectively. The tolerance of MMF was studied prospectively with a follow-up of 1.1+/-0.2 years. Gastrointestinal disorders requiring dosage reduction or discontinuation of therapy, observed in five of nine patients, occurred at an incidence higher than expected from adult data. Our results suggest that the dose of 600 mg/m(2) every 12 h extrapolated from adult data for use in pediatric patients would be associated with plasma levels and AUCs higher than expected and may be associated with a higher incidence of side-effects, primarily gastrointestinal. PMID- 10684356 TI - Pharmacokinetics of mycophenolate mofetil are influenced by concomitant immunosuppression. AB - The recommended dosage for mycophenolate mofetil (MMF) in combination with cyclosporin (CyA) for pediatric kidney transplant recipients is 600 mg/m(2) twice daily (b.i.d.). We recently published pharmacokinetic (PK) profiles of MMF in combination with tacrolimus (FK506): in order to keep the mycophenolic acid (MPA) pre-dose trough concentration between 2 and 5 microg/ml and to avoid side effects, mean MMF doses were reduced to 300 mg/m(2) b.i.d. In order to investigate whether this striking difference was due to alterations of MPA clearance by CyA or FK506, we analyzed PK profiles from 13 patients who received MMF without CyA or FK506, and compared these data with 14 patients who received a combination of MMF and FK506 and 15 patients who received MMF and CyA. Mean area under the curve (AUC) in all PK profiles was 61.9+/-23.8 microgxh/ml. Although the AUCs did not differ between the groups, the dose per square meter was significantly lower in patients receiving concomitant FK506 compared with CyA, and the dose-normalized AUC was significantly higher. The MMF doses were 1,158+/ 301 mg/m(2) per day in the CyA group, 555+/-289 mg/m(2) per day in the tacrolimus group, and 866+/-401 mg/m(2) per day in the group without concomitant calcineurin inhibitor treatment. The apparent clearance of MPA is reduced in combination with tacrolimus. The reason for this remains unknown. There was a trend towards lower dose-normalized AUCs in the CyA group compared with the group without calcineurin inhibitor treatment. We conclude that concomitant medication alters the clearance of MPA. It is noteworthy that there was substantial interindividual variation, despite the rather marked differences between the groups, and therefore we recommend starting MMF in combination with CyA at a dose of 600 mg/m(2) b.i.d., in combination with tacrolimus at a dose of 300 mg/m(2) b.i.d., and without a calcineurin inhibitor at a dose of 500 mg/m(2) b.i.d., and adjusting doses using therapeutic drug monitoring of MPA. PMID- 10684357 TI - Risk factors for hyperlipidemia in long-term pediatric renal transplant recipients. AB - Hyperlipidemia (HL) is a common problem in adult renal transplant (TP) recipients, contributing to an increased risk of cardiovascular disease and chronic TP nephropathy. There are multiple causes of HL post renal TP in adult patients, including pre TP HL, immunosuppressive agents, renal dysfunction, hypoalbuminemia secondary to nephrotic syndrome, obesity, and conditions that lead to end-stage renal disease (ESRD). We evaluated the incidence and risk factors of HL in 62 pediatric renal TP recipients (15.4+/-4.2 years, range-3.0 22.3 years) with long-term (6.7+/-3.1 years) functioning [glomerular filtration rate (GFR) 66.7+/-23.2 ml/min per 1.73 m(2)] allografts. The mean serum cholesterol (C) level was 205. 5+/-43.6 mg/dl. Thirty-two patients (51.6%) exhibited elevated serum C levels. The mean serum triglyceride (TG) level was 157.3+/-88.4 mg/dl. Serum TG levels were elevated in 32 patients (51.6%). In patients with elevated serum levels of either C or TG, the mean low-density lipoprotein level (LDL) was 138.6+/-44.1 mg/dl (normal <130 mg/dl) and the high density lipoprotein (HDL) level 54.6+/-15.9 mg/dl (normal>34 mg/dl). Of those patients studied, 45.5% had high LDL levels, whereas 9.1% exhibited low HDL levels. The two risk factors for elevated serum C levels in our patient population were pre-TP HL and increased years since TP. The only risk factor for elevated serum TG levels was reduced GFR. A family history of HL had a significant deleterious impact upon serum levels of C (P=0.01), but did not affect serum TG levels (P=0.7). Years on dialysis prior to TP, history of prior TP, gender, body mass index, and disease leading to ESRD had no influence upon the development of post-TP HL. We conclude that post-renal TP HL is a significant problem in pediatric renal TP recipients. PMID- 10684358 TI - Fatal disseminated Nocardia farcinica infection in a renal transplant recipient. AB - Six years after a renal cadaver transplant, a 20-year-old girl developed multiple painful cutaneous abscesses and bilateral pneumonia secondary to Nocardia farcinica infection. Despite broad in vitro sensitivity to several antibiotic agents and aggressive medical treatment, the patient failed to respond and died after 10 weeks of therapy. We conclude that Nocardia farcinica is a very aggressive organism in immunocompromised patients and is often resistant to antimicrobial agents. PMID- 10684359 TI - Reduction of peritonitis with the rectus abdominis muscle flap in a CAPD patient. AB - An adolescent maintained on continuous ambulatory peritoneal dialysis (CAPD) for 8 years had relapsing peritonitis involving peritoneal catheter tunnel infections. We attempted catheter removal and replacement simultaneously, with the catheter covered cylindrically by a rectus abdominis muscle flap to prevent recurrent tunnel infections. During 3 years of follow-up, there have been no episodes of peritonitis involving tunnel infection. Our modified insertion technique can eradicate tunnel infection, thus reducing peritonitis. PMID- 10684360 TI - Mutational analysis of COL4A5 gene in Korean Alport syndrome. AB - Mutational analysis of the COL4A5 gene in X-linked Alport syndrome (AS) requires an expensive and time-consuming procedure with a detection rate of 50%, at best. There have been three multicenter collaborative studies of mutation analysis in the COL4A5 gene using systematic screening of entire coding regions of the gene. This is a similar study executed in a single center in Korea. Twenty-five unrelated Korean patients with AS in whom the diagnosis was confirmed pathologically were included in the study. By systematic screening of all 51 exons of the gene using polymerase chain reaction/single-strand conformation polymorphism analysis, ten mutations were detected in 10 unrelated patients. These included one medium-sized deletion involving exon 49-51, one single base pair deletion, one nonsense point mutation, one splice site mutation, and six missense point mutations. Of the six missense mutations, four involved a glycine residue and disrupted the Gly-X-Y repeats in the collagenous domain. The overall detection rate of mutations was 40%. Although DNA analysis in AS is currently not applicable to routine clinical diagnosis due to several practical and technical problems, it is likely to replace morphological diagnosis in the near future. PMID- 10684361 TI - Acidosis and weight loss are induced by cyclosporin A in uninephrectomized rats. AB - The effects of cyclosporin A (CyA, 50 mg/kg body weight) or its commercial vehicle (cremophor) on the acid-base regulation of uninephrectomized rats were assessed for 7 days and in non-nephrectomized rats for 15 days. CyA induced a marked systemic acidosis, accompanied by decreases in blood PCO(2) and plasma bicarbonate. Untreated uninephrectomized rats did not show the acidosis. In CyA treated rats the urine pH decreased (control 6. 65+/-0.06 vs. CyA 6.18+/-0.08; P<0.01) as well as urinary bicarbonate (non-nephrectomized rats 7.50+/-1.88 mM vs. uninephrectomy plus CyA 0.75+/- 0.06 mM; P<0.01), suggesting partial renal compensation of systemic acidosis. Titratable acidity increased in CyA-treated rats (control 21.6+/-1.2 vs. CyA 63.3+/-12.0 microEq/l; P<0.001). Phosphate, glucose, and osmolar clearances were not significantly altered in non nephrectomized rats treated with CyA for 15 days. There was a striking decrease in body weight in CyA-treated rats (control 274.0+/-3.8 vs. CyA 225.0+/-5.1 g; P<0. 01), but compensatory growth of the remaining kidney was not prevented by this drug or by its vehicle. In summary, CyA induced a severe metabolic acidosis in uninephrectomized rats that was not compensated by the remaining kidney, in spite of the well-preserved compensatory weight gain of this organ. Loss of body weight was significant in CyA-treated animals. PMID- 10684362 TI - Phospholipase A(2) activity, heat shock protein, and superoxide dismutase in rat remnant kidney. AB - Male Sprague-Dawley rats (150-200 g) were randomly assigned to sham operation (n=6) or 5/6 nephrectomy (n=12) procedures. Two weeks after the completion of the 5/6 nephrectomy, these animals were again randomly assigned to two groups: non treatment or treatment with vitamin E supplementation at 200 IU/kg chow. Two weeks later, all animals were sacrificed and the kidneys harvested. The secretory phospholipase A(2) (PLA(2)) activity was elevated (150%) in the untreated remnant kidney but returned to sham values in the vitamin E-treated kidneys. The cytoprotective heat shock protein (HSP70) and the intracellular antioxidant superoxide dismutase (MnSOD, Cu/ZnSOD) were similar in sham, remnant, and vitamin E-treated remnant kidneys. We conclude that the sudden reduction of renal mass secondary to the 5/6 nephrectomy procedure stimulates PLA(2) activity but not HSP70, MnSOD, or Cu/ZnSOD. This increased activity of PLA(2) in the remnant kidney returned to sham values after vitamin E treatment. The intrinsic cellular antioxidant enzymes, MnSOD, Cu/ZnSOD, as well as the cytoprotective heat shock protein HSP70, showed no significant changes in either vitamin E-treated or untreated kidneys compared with sham. These data are suggestive that the elevation of PLA(2) is a specific and localized response to the sudden reduction of renal mass. PMID- 10684363 TI - Idiopathic collapsing glomerulopathy in children. AB - Idiopathic collapsing glomerulopathy (ICG) is a clinically and pathologically distinct variant of focal segmental glomerulosclerosis, characterized clinically by rapid progression of renal insufficiency, a male and African-American racial predominance, and pathologically by segmental glomerular collapse, visceral epithelial cell hypertrophy and hyperplasia, and the absence of endothelial tubuloreticular inclusions. Pathologically similar lesions have been reported in adult and pediatric patients with human immunodeficiency virus (HIV) infection and/or intravenous (IV) drug abuse. Most patients with ICG who have been reported in the literature are adults. Six children with ICG were retrospectively identified (two from East Carolina University, four from University of North Carolina-Chapel Hill). Clinical data and renal biopsy findings were reviewed for all patients. All six patients were male; five African-American and one Hispanic. Ages ranged from 2 to 17 years (mean 12 years). Steroid-resistant nephrotic syndrome was the presenting clinical finding. Average 24-h urine protein excretion was 6.3 g (range 3.2-15 g). Five patients were serologically negative for HIV infection (one patient not tested) and none had a history of IV drug abuse or known HIV risk factors. Progression to end-stage renal insufficiency in two patients within 1 year of biopsy required renal transplantation, and within 1 month of biopsy one patient required dialysis. We report a series of pediatric patients with ICG, an aggressive variant of focal segmental glomerulosclerosis. ICG in children is similar clinically and pathologically to this disease in adult patients. PMID- 10684365 TI - Bartter syndrome in a neonate: early treatment with indomethacin. AB - The neonatal form of Bartter syndrome is characterized by intrauterine onset of polyuria leading to severe polyhydramnios. We report a patient with the early onset of the syndrome and a similar history in a previous sibling who died in early neonatal life. The patient is a female product of 33 weeks of gestation complicated by severe polyhydramnios. Her birth weight was 2,100 g. Polyuria led to severe dehydration on the 3rd day of life. Laboratory studies showed hypokalemia, hyponatremia, and elevated plasma levels of renin and aldosterone. Hypercalciuria was associated with echographic evidence of nephrocalcinosis. Indomethacin therapy resulted in a significant reduction in urine volume and correction of biochemical abnormalities. Growth and development are satisfactory after 4 years of indomethacin therapy, but nephrocalcinosis remains unchanged. PMID- 10684364 TI - Hemostatic problems and thromboembolic complications in nephrotic children. AB - A hypercoagulable state and the risk of thromboembolism in both arterial and venous circulation is a relatively frequent and serious feature of nephrotic syndrome (NS) in children and adults. The aim of this study was to evaluate the coagulation states of children with NS before and after corticosteroid (CS) therapy and to compare the results with a healthy control group. The first group consisted of 49 nephrotic children (30 boys and 19 girls) with a mean age of 6. 5+/-4.9 years (range 1-16 years). The control group included 17 healthy children (9 boys and 8 girls). At the time of admission, all patients were evaluated for the presence of clinical thromboembolism, hematological and biochemical indicators of a hypercoagulative state, and renal disease. This was repeated after CS treatment. Deep vein thrombosis was observed in 2 nephrotic patients who had very low plasma antithrombin III (AT III) levels and fibrinogen levels above 750 mg/dl. Thus, the prevalence of thromboembolism was 4% in our pediatric nephrotic population. The mean AT III level of the study group was 68.2+/-23.4% at the onset of the disease, which was significantly lower than the level of the control group (84.0+/-7. 6%). Plasma AT III levels increased to 74.4+/-15.3% after CS treatment, which correlated with the serum albumin levels. However, there was no correlation with urinary protein excretion. Protein C levels were higher than controls during all stages of the disease in both steroid-responsive and -unresponsive patients. The mean protein S level was similar in both groups. Plasma fibrinogen and cholesterol levels were significantly higher in the study group but decreased to within normal limits with remission. Our study suggests that thromboembolic complications are not infrequent in children with NS, and may be related to low plasma AT III and albumin and high fibrinogen and cholesterol levels. PMID- 10684366 TI - Vesicoureteric reflux associated with renal dysplasia in the Wolf-Hirschhorn syndrome. AB - Wolf-Hirschhorn syndrome (WHS) is caused by a partial deletion of the short arm of chromosome 4 (4p16.3) and is characterized by severe pre- and postnatal growth retardation, developmental delay, and multiple congenital anomalies, including malformations of the urogenital system. We describe the renal and urinary tract phenotype in a series of six children with WHS. Vesicoureteric reflux was present in four of our six patients (5 of 10 ureters), an abnormality not previously reported in WHS. PMID- 10684367 TI - A new variant of apolipoprotein E (apo E Maebashi) in lipoprotein glomerulopathy. AB - We have previously reported an 8-year-old girl with lipoprotein glomerulopathy. Assessment of serum apolipoprotein E (apo E) in this patient showed a discrepancy between phenotype and genotype, suggesting that she may have a variant of apo E. The present report concerns our analysis of DNA sequences of the apo E gene in the patient: nine base pairs were found to be deleted from exon 4. This mutation would appear to encode a new apo E variant lacking three amino acids. This variant may be associated with the pathogenesis of lipoprotein glomerulopathy. PMID- 10684368 TI - Gene expression after intrarenal injection of plasmid DNA in the rat. AB - Effective gene therapy requires efficient delivery and expression of the necessary genetic information to the target tissue. We demonstrate here that plasmid DNA, injected as naked, uncomplexed DNA into the cortical region of rat kidney, or intravenously, is localized and expressed in the kidney. The plasmid pRSVZ contained the Rous sarcoma virus promoter and a reporter gene, the beta galactosidase gene, derived from bacteria. The beta-galactosidase gene hydrolyzes the artificial substrate X-gal to produce an intense blue color in cells that have taken up and expressed the plasmid genes. We have used X-gal staining and Western blotting to study plasmid gene expression 1, 4, and 8 days and 6 months after intrarenal injection of 50 microg of plasmid DNA and at 1 and 4 days after intravenous injection. Expression was apparent in the kidneys and several other tissues 24 h after injection and persisted for at least 8 days; expressed proteins could still be detected in the injected kidney 6 months later. These observations were corroborated by use of a plasmid, pEGFP-Puro, harboring the cytomegalovirus promoter in conjunction with a different reporter gene, the green fluorescent protein (GFP). Histological localization and Western blotting analysis of GFP expression after intrarenal injection of pEGFP-Puro paralleled results obtained with the plasmid pRSVZ. Our findings support the suggestion that intrarenal or intravenous injection of naked plasmid DNA may be an effective means of delivering therapeutic genes to the kidney and several other tissues. PMID- 10684369 TI - Treatment of lupus nephritis in children. AB - In children, systemic lupus erythematosus (SLE) is often more severe than in adults. Renal disease is very common in SLE, with clinical symptoms of renal involvement occurring in 30%-70% of patients. In the absence of appropriate treatment the child may die from the disease or progress rapidly to renal failure. However, aggressive treatment regimens, in particular corticosteroids, carry the risk of growth retardation, accelerated atherosclerosis, and severe infectious complications. Lupus nephritis is classified into six groups depending on the severity of the histological lesions. The most-appropriate treatment for optimal efficacy with minimal side-effects depends on the disease severity. Mild lesions (class I or II) require only careful follow-up to identify any disease progression. Patients with class III nephropathy (focal and segmental glomerulonephritis) may have mild clinical symptoms, in which case no specific therapy is indicated, or more-severe symptoms of the nephrotic syndrome, hypertension, and sometimes moderate renal insufficiency. These patients require the same aggressive therapy as those with class IV disease (diffuse proliferative glomerulonephritis). Our current protocol starts with three methylprednisolone pulses followed by 1.5 mg/kg per day oral prednisone and six monthly pulses of cyclophosphamide. After a second renal biopsy the patient may be maintained on azathioprine while the prednisone dosage is slowly tapered. In children with milder disease we use lower doses of oral prednisone (1-1.5 mg/kg per day). Patients with membranous glomerulonephritis (class V) require no specific therapy if they have pure membranous nephropathy, but require aggressive therapy if they have the nephrotic syndrome. In those patients who progress to end-stage renal disease, clinical and serological remission is common and renal transplantation can be performed, as recurrence in the transplant is very rare. PMID- 10684370 TI - Fetal therapy for obstructive uropathy: past, present.future? AB - Antenatal treatment of obstructive uropathy, although widely performed, remains controversial. An overview of prenatal therapy for obstructive uropathy, the limitations of the early published experience, advances of recent years, and future directions for treatment are reviewed. The clinical approach and outcomes of the Fetal Treatment Program of Hutzel Hospital and Wayne State University are presented. Patient selection for antenatal treatment is based on the existence of a significant threat of neonatal death due to pulmonary hypoplasia, pending exclusion criteria such as anatomical structural anomalies and chromosomal defects. Ultrasonography, karyotyping, and sequential urinary electrolyte analysis are essential. Current treatment involves the placement under ultrasonic guidance of a Rodeck vesicoamniotic shunt. Recent technical advances include the use of amnioinfusion for fetal visualization, temporary fetal paralysis, routine antibiotics, and more-precise catheter placement. The establishment of standardized short- and long-term outcome measures and the documentation of fetal and maternal complications are in progress. Procedural refinement, development of fetoscopic techniques and equipment, identification of urinary markers to aid patient selection, and the collection of multicenter outcome data will assist the future management of prenatally detected obstructive uropathy. PMID- 10684371 TI - Ask the expert. PMID- 10684372 TI - IPNA newsletter PMID- 10684373 TI - Histologic examination of bone marrow core biopsy specimens has limited value in the diagnosis of mycobacterial and fungal infections in patients with the acquired immunodeficiency syndrome. AB - Bone marrow cultures and biopsy specimens are commonly obtained to rule out disseminated infections, especially in persons with the acquired immunodeficiency syndrome (AIDS) and cytopenias. Using culture as the gold standard, we reviewed 130 consecutive bone marrow cores obtained from 114 AIDS patients along with results of concurrent blood and/or bone marrow aspirate cultures to determine the usefulness of histologic examination for diagnosis of mycobacterial and fungal infections. We also compared the ability of Ziehl-Neelsen, auramine-rhodamine (AR), polyclonal antibody to Mycobacterium bovis (Ab), and Gomori's methenamine silver staining to detect infections. Twenty-seven patients had mycobacterial infection (25 Mycobacterium avium-intracellulare complex cases and two Mycobacterium tuberculosis cases) detected by blood and/or bone marrow cultures. The maximum sensitivity of histology was 50% when the auramine-rhodamine stain and the polyclonal antibody to M bovis were used in combination. The single best stain was auramine-rhodamine, with a sensitivity of 44%, followed by the polyclonal antibody to M bovis (35%). Granulomas were observed in nine cases of mycobacterial infection and did not correlate with the presence of stainable organisms. Of seven patients with positive fungal cultures of bone marrow, four had granulomas and a positive Gomori's methenamine silver stain, one had only a positive stain, and two had neither granulomas nor a diagnostic stain. Overall, granulomas were not sensitive for the detection of infections when culture-proven mycobacterial and fungal cases were evaluated together. We conclude that bone marrow examination has a limited value in the routine evaluation of common opportunistic infections in AIDS patients and recommend that less-invasive tests, such as blood cultures, be obtained initially in most circumstances. PMID- 10684374 TI - Epstein-Barr virus in squamous carcinoma of the anterior nasal cavity. AB - Squamous carcinoma is the most common malignancy of the head and neck, but it rarely occurs in the nasal vestibule. Epstein-Barr virus (EBV) has been detected in and is causally linked to various head and neck tumors, particularly nasopharyngeal carcinoma. The possible role of EBV in squamous carcinoma of the anterior nasal cavity, particularly of the nasal vestibule, has not been previously investigated. Histologic sections from 17 patients with nasal vestibular squamous carcinoma were examined. Material for EBV detection by immunohistochemistry and by in situ hybridization was available in 15 of the 17 cases. The study group consisted of eight men and nine women ranging in age from 40 to 82 years (mean age, 64 years). None of the patients was of Asian descent. The squamous carcinomas were graded as well differentiated (one case), moderately differentiated (11 cases), and poorly differentiated (five cases). Fourteen patients were smokers; the history of smoking ranged from 20 to 60 pack-years. Treatment modalities included surgical resection, radiation, chemotherapy, or a combined approach. The clinical follow-up periods ranged from 7 months to 16 years. Three patients developed metastases, one of whom died of disease after 1 year. Epstein-Barr virus was not detected in any of the 15 of 17 cases tested by either immunohistochemistry or by in situ hybridization. Squamous carcinoma of the nasal vestibule is an uncommon cancer that is not causally related to EBV. PMID- 10684375 TI - Prognostic significance of nuclear DNA content in chondrosarcoma. AB - Chondrosarcoma is the second most frequent primary malignant tumor of bone. Many of these tumors represent histopathologic borderline cases. In this study, DNA ploidy status, 2c deviation index (2cDI), and DNA malignancy grade (DNA-MG; based on the variation of nuclear DNA content of tumor cells around the normal DNA [2c] peak) were examined for their diagnostic and prognostic value in comparison with conventional histopathologic grading. Twenty-two paraffin-embedded samples were available for histopathologic investigation and for quantitative cytophotometric DNA determination of Feulgen-stained nuclei. Clinicopathologic parameters and prognosis were analyzed over a maximum follow-up period of 252 months. Nineteen of 22 (86%) chondrosarcomas showed aneuploid DNA content. 2cDI (r =.58, P <.01) and DNA-MG (r =.58; P <.01) correlate with the histopathologic grading. Significant correlation between the 2cDI (P <.01) and DNA-MG (P <.025) and the overall survival was found. Ploidy did not influence the overall survival rate. In metastasis-free patients, the 2cDI and DNA-MG gave better prognostic information than conventional histopathologic grading. When patients developed metastasis, however, histopathologic grading was the prognostic parameter of choice. Cytometric DNA measurement provide additional objective information regarding the diagnosis and prognosis of chondrosarcomas, even more than that obtained by conventional histopathologic grading, and may be helpful in planning the treatment. PMID- 10684376 TI - Fine-needle aspiration of granulocytic sarcomas: a morphologic and immunophenotypic study of seven cases. AB - Granulocytic sarcoma is an uncommon extramedullary, solid tumor of myeloid cells. Only rarely has this entity been diagnosed by fine-needle aspiration (FNA) cytology. This report encompasses the cytologic findings of FNAs from seven patients with granulocytic sarcomas, including four male and three female patients with a mean age of 52 years (range, 12 to 77 years). The aspirates were obtained from skin or subcutaneous tissue (four cases), testis (one case), posterior ileum (one case), lymph node (one case), and abdominal washing (one case). Morphology of the aspirates varied from well-differentiated to poorly differentiated cells showing little or no evidence of myeloid differentiation. Thorough search for evidence of myeloid differentiation and a high index of suspicion of granulocytic sarcoma are of paramount importance. In three cases, flow cytometric and immunocytochemical studies were applied to the FNA materials to confirm the myeloid lineage of the cells and the diagnosis. In the other four cases more than one site was involved by the tumor; once the diagnosis of granulocytic sarcoma was established with a biopsy, the FNA sufficed to confirm the diagnosis at another location. This study demonstrates that FNA cytology in conjunction with appropriate immunophenotyping can provide an accurate diagnosis of granulocytic sarcoma. Fine-needle aspiration can reduce the need for surgical intervention when combined with immunophenotypic studies and when additional anatomic sites are involved. PMID- 10684377 TI - Isolated Langerhans cell histiocytosis of the thyroid: a report of two cases with nuclear imaging-pathologic correlation. AB - We describe two cases of isolated langerhans cell histiocytosis (LCH) of the thyroid gland, one of which was found in conjunction with an incidental papillary carcinoma. The first case was that of a 43-year-old man who presented with a 1- to 2-cm nodule within the left lobe of the thyroid. Fine-needle aspiration cytology revealed atypical cells with convoluted nuclei in a background of eosinophils and lymphocytes. The findings prompted a recommendation for excision secondary to the high suspicion of a hematologic malignancy. Histologic sections demonstrated LCH in association with a small focus of papillary carcinoma. The second case involved a 43-year-old woman who presented with a 1.8-cm nodule within the right lobe of the thyroid. Fine-needle aspiration in this case demonstrated abundant hemosiderin-laden macrophages, occasional lymphocytes, and a single benign sheet of follicular cells. No eosinophils were seen; however, a single group of atypical histiocytic cells with cleaved nuclei was noted. The nodule was subsequently resected. Histologic examination demonstrated LCH in association with follicular nodular hyperplasia with cystic degeneration. Immunohistochemical studies were performed in both cases, revealing CD1a and S100 immunoreactivity in the Langerhans' cells. Although LCH may occur as a manifestation of systemic disease, its occurrence as an isolated finding in the thyroid is rare. Its occurrence in association with papillary carcinoma of the thyroid is even more uncommon. We present two cases of isolated LCH of the thyroid, one of which was found in association with papillary carcinoma of the thyroid. The cytologic, histologic, immunohistochemical, and radiologic features are described in each case. The ultrastructural findings from the first case are also presented. PMID- 10684379 TI - Ossifying well-differentiated Sertoli-Leydig cell tumor of the ovary. AB - A unique case of an ovarian sex cord-stromal tumor occurring in a pregnant 20 year-old is described. The tumor showed central ossification on macroscopic examination. Microscopically, cords and nests of Sertoli cells were identified, mostly away from the abundant central hyalinization, calcification, and ossification. A small number of Leydig cells were present, with isolated Reinke crystals. The presence of these cells could reflect luteinized stromal cells secondary to pregnancy. The Sertoli cells were dominant and the calcified/ossified areas were at the center of a dominant Sertoli nodule. This degree of ossification has never been reported in either ovarian Sertoli tumors or well-differentiated Sertoli-Leydig tumors. Calcifying Sertoli cells neoplasms have been described in the testis, but this case appears to be the first description of a case with similar features in the ovary. PMID- 10684378 TI - Endometrioid carcinoma arising in pericecal endometriosis clinically mimicking Crohn's disease. AB - A case of endometrioid carcinoma arising in pericecal endometriosis that clinically and radiologically mimicked Crohn's disease is presented. After developing several complications of steroid therapy for presumed Crohn's disease, a 48-year-old woman developed intestinal obstruction and underwent a right hemicolectomy. A pericecal mass composed of endometriosis and endometrioid carcinoma and a locally metastatic ileal carcinoid tumor were resected. The patient recovered fully and is clinically free of tumor at 36 months. The pertinent literature is reviewed and the etiologic, therapeutic, and prognostic implications of this case are discussed. PMID- 10684380 TI - A report of mesothelial/monocytic incidental cardiac excrescences and a literature review. AB - We report the case of a rare cardiac lesion, mesothelial/monocytic incidental cardiac excrescences, and also provide a review of the literature. Diagnosis of this entity was based on both its unique morphologic features and imunohistochemical stains. Cytokeratin positivity confirmed the epithelial component, mesothelial cells, in the lesion. Positive staining of CD68 in the monocytic-appearing cells revealed the histiocytic nature of the second component of this lesion. Differential diagnoses are discussed. This report emphasizes the diagnostic dilemma encountered with this unusual entity and the possibility of misdiagnosing the epithelial portion as a metastatic lesion or vice versa. PMID- 10684381 TI - A simple yet effective technique to improve laboratory safety for the grossing of large surgical specimens. AB - In this article we describe a simple technique to avoid accidential needle stick injuries while grossing large specimens. Rubber bands instead of needles and Plexiglas instead of wax board is used to eliminate needle stick injuries. This method is quite simple and imprints of rubber bands do not interfere with gross anatomy of the specimen. PMID- 10684382 TI - The pathology of acute appendicitis. AB - Although acute appendicitis is frequent, it is subject to common misconceptions. Furthermore, there is little good evidence to support some of our beliefs. This report reviews the role of the anatomic pathologist in diagnosis when acute appendicitis is suspected clinically and discusses what is known of its pathology. The conclusions that can be legitimately drawn from the literature are emphasized. A classification is proposed that incorporates intraluminal inflammation, acute mucosal inflammation, acute mucosal and submucosal inflammation, suppurative (phlegmonous) appendicitis, gangrenous appendicitis, and periappendicitis, and the significance of each of these diagnoses is discussed. The etiology and pathogenesis of acute appendicitis is reviewed. Contrary to popular belief, the best evidence indicates that obstruction is unlikely to be the primary cause, at least in the majority of cases. Ancillary techniques in the diagnosis of appendicitis, including laparoscopy and peritoneal aspiration cytology, are discussed. PMID- 10684384 TI - Local lymph node assay: validation assessment for regulatory purposes. AB - For the prediction of skin sensitization potential of substances, the murine local lymph node assay (LLNA) is an alternative to the widely used guinea pig tests. For more than 10 years, this method has undergone extensive development, evaluation, and validation. In this review, the validation status of the LLNA is considered, specifically with regard to its use for regulatory identification of skin sensitization hazards. The LLNA is a method for the predictive identification of chemicals that have a potential to cause skin sensitization. Activity is measured as a function of lymph node cell proliferative responses stimulated by topical application of test chemicals. The LLNA has successfully passed all reasonable validation stages. It provides a reliable and relevant source of predictive skin sensitization data, which unlike results from guinea pig tests, are reproducible from laboratory to laboratory. In summary, the LLNA is now ready for acceptance as a viable and complete alternative to traditional methods, offering a substantial reduction in animal numbers and refinement opportunities without compromising the standards for the identification of important skin sensitizers. PMID- 10684385 TI - Skin aging and photoaging: the role of DNA damage and repair. AB - Both genetic (intrinsic) and environmental (extrinsic) factors contribute to the phenotypic changes in cutaneous aging. However, only recently have the underlying molecular mechanisms involved in these changes been elucidated. DNA damage to both genomic and mitochondrial DNA and subsequent DNA repair contribute greatly to age-associated skin changes and carcinogenesis. Better understanding of these intricate, interwoven mechanisms involved in DNA damage and repair might help to develop new strategies in preventing and treating changes of intrinsic skin aging and photoaging, improving skin appearance and reducing the risk of skin cancer. PMID- 10684386 TI - Textile dermatitis in Israel: a retrospective study. AB - BACKGROUND: The diagnosis of contact dermatitis caused by clothing may be difficult because of its clinical polymorphism. Data in the literature suggest that textile dermatitis is more common than previously thought. OBJECTIVE: Our purpose was to study our patients suspected of having textile contact dermatitis from 1991 to 1997. METHODS: The records of the patients with positive reactions to allergens from the Textile Colors and Finish series in 3 contact dermatitis clinics were reviewed. All the patients were clinically evaluated and patch tested with the European Standard series and the Textile Colors and Finish series (Chemotechnique Diagnostics, Malmo, Sweden). RESULTS: Twenty-two of the 55 patients (40%) had positive patch tests to the textile dye allergens. Four of them had occupationally related textile dermatitis. The most frequent allergens were Disperse Blue 124, Disperse Blue 85, Disperse Red 17, and Disperse Blue 106. Erythematosquamous lesions were the most common forms of textile dermatitis (56%), followed by pustular lesion (16%) and hyperpigmented patches (8%). CONCLUSIONS: The relatively high percentage of positive results (40%) was attributable to the selected cohort of patients. In our series, positive reactions to the allergens Disperse Blue 124, 85, and 106 were common findings. Clinically, pustular allergic contact dermatitis, triggered by textile dyes was observed along with the more frequent erythematosquamous clinical form. PMID- 10684387 TI - Disperse blue dyes 106 and 124 are common causes of textile dermatitis and should serve as screening allergens for this condition. AB - BACKGROUND: Textile dye dermatitis is frequently undiagnosed because clinical awareness is low and because of the absence of good screening allergens in standard patch test series for this type of contact dermatitis. OBJECTIVES: To determine the incidence of textile dye allergy in patients with problematic eczemas evaluated at a contact dermatitis clinic, and to determine the incidence of allergic contact dermatitis to diperse blue dyes in these patients. METHODS: We conducted a retrospective study of 788 patients who were patch tested to either the North American Contact Dermatitis Group (NACDG) Standard Series or the European Standard Series, in addition to other relevant series. The Chemotechnique textile series was utilized in 271 patients (28%). RESULTS: Forty patients reacted positively to 1 or more textile dyes, the majority reacting positively to Disperse Blue 106 (33 of 40; 82.5%) and to Disperse Blue 124 (32 of 40; 80%). Ten of 11 tested patients reacted to their own clothing, 9 of whom reacted to the blue/black 100% acetate or 100% polyester liners in their garments. CONCLUSIONS: Textile dye allergy is more common than previously reported. It can cause marked dermatitis and widespread autoeczematization reactions. The most frequent allergens are Disperse Blue 106 and 124, which are frequently found in the 100% acetate and 100% polyester liners of women's clothing. We recommend that Disperse Blue 106 or 124 serve as the screening allergen for textile dye dermatitis. PMID- 10684388 TI - Cross-reactivity patterns of cobalt and nickel studied with repeated open applications (ROATS) to the skin of guinea pigs. AB - BACKGROUND: The relevance of patch-test reactivity to chemicals on cross challenge is hard to state, but it is generally assumed that the patient might risk a relapse of contact dermatitis when exposed to the cross-reacting compound(s). OBJECTIVE: To study relevance by using the repeated open application test (ROAT) and applying the inducing allergen cobalt chloride (CoCl(2)) or nickel sulfate (NiSO(4)) as well as the possibly cross-reacting compound (NiSO(4) or CoCl(2)) topically to guinea pigs. METHOD: Animals were induced according to the guinea pig maximization test (GPMT) method, patch tested and then treated for 10 days using ROATs. Sensitivity thresholds were determined with serial dilution tests. RESULTS: Guinea pigs induced with CoCl(2) reacted in patch testing (100%) and in ROATs to CoCl(2) (93%) but not to NiSO(4). Animals induced with NiSO(4) reacted in patch testing to NiSO(4) (100%) but not to CoCl(2), and in the ROATs to NiSO(4) (41%) and less to CoCl(2). CONCLUSIONS: Our results support the assumption that the concomitant patch test reactivity is due to multiple sensitizations rather than cross-reactivity. We previously found that animals induced with palladium chloride (PdCl(2)) also reacted to NiSO(4) on patch testing but not in the ROATs, indicating that the results from patch testing might overestimate the risk of a relapse. ROATs in patients with solitary and/or concomitant sensitivity to CoCl(2), NiSO(4) or PdCl(2) are desirable. PMID- 10684389 TI - Symptoms and signs reported during patch testing. AB - BACKGROUND: In a pilot questionnaire study, there was a high frequency of subjective complaints and distant skin reactions during patch testing as reported at the day of test reading, particularly in female patients. OBJECTIVE: To document in a controlled study possible side-effects of a generalized nature occurring during the test procedure. METHODS: A questionnaire study on symptoms and signs reported at application and at reading of standard patch tests was conducted with 401 patients, with the patients serving as their own controls. RESULTS: An eczematous flare-up during patch testing was observed in 3.7% of the patients. There were plenty of different symptoms of malaise but, with one exception (itch on the back), the number of symptoms tended to be less on the day of reading than on the day of application of the tests. This held true also for itch occurring in the patients' dermatitis. There was no statistical correlation between symptoms and signs on the one hand and positive patch tests on the other. CONCLUSION: Distant skin reactions and impairment of general health occurring during patch testing are often reported at the time of test reading. However, with the exception of itch on the back, symptoms and signs are rather less common after the application of patch tests than before. PMID- 10684390 TI - Allergic contact dermatitis caused by parabens: 2 case reports and a review. AB - Parabens, methyl, ethyl, propyl, benzyl, and butyl, are the most common preservatives in use today. They are the alkyl esters of p-hydroxybenzoic acid and are used extensively because they are relatively nonirritating and nontoxic and offer good antimicrobial coverage. Testing for paraben allergen can be done by patch testing. Two cases of allergic contact dermatitis (ACD) to parabens are used to discuss the background of parabens, their allergenicity, patch testing issues, and several "paraben paradoxes." Although ACD to parabens has been reported, given the widespread use, it is relatively uncommon. Because of their low rate of allergenicity and their favorable preservative profile and efficacy, parabens remain the number one preservative in use. PMID- 10684391 TI - Phototesting and photopatch testing: when to do it and when not to do it. AB - Phototesting and photopatch testing are among the most important tests in the evaluation of photodermatoses, yet their use has been restricted to specialized centers. To assist clinicians interested in conducting these procedures and in updating their techniques, we asked 4 experts to comment on these tests in the context of diagnostic approach to the photosensitive patient. A list of photoallergens and a protocol for their use is provided. PMID- 10684392 TI - Why a New Journal in Cardiovascular Pharmacology and Therapeutics? PMID- 10684393 TI - Antiarrhythmic Therapy of Ventricular Arrhythmias: The Contemporary Dilemma. PMID- 10684394 TI - Effects of Sotalol on His-Purkinje Conduction and Refractoriness in Humans. AB - BACKGROUND: The effects of sotalol on refractoriness in human ventricular and atrial muscles have been well established, but the drug's effect on the electrical properties of the His-Purkinje system in humans is not known, especially whether sotalol's effect is due solely to its action on prolonging repolarization or in combination with its beta-blocking properties. We studied the electrophysiologic effects of intravenous sotalol and propranolol in patients undergoing electrophysiologic studies of cardiac arrhythmias. METHODS AND RESULTS: We studied 22 patients (19 men, 3 women; mean age, 60 +/- 6 years) who had coronary artery disease and assessable anterograde, retrograde, or both, His Purkinje system function. Fifteen patients underwent electrophysiologic studies before and after intravenous sotalol (1.5 mg/kg), and 7 patients underwent electrophysiologic studies before and after intravenous propranolol (0.15 mg/kg). Both sotalol and propranolol had no significant effect on the H-V interval, but sotalol significantly increased ventricular refractoriness and His-Purkinje refractoriness, both in anterograde and retrograde conduction, whereas propranolol did not, Sotalol's effect on His-Purkinge refractoriness also caused atrioventricular block distal to the His bundle during atrial pacing at a moderately fast rate. Sotalol was not effective in preventing bundle branch re entry tachycardia, nevertheless, it increased cycle length of bundle branch re entry tachycardia by increasing refractoriness. CONCLUSIONS: Sotalol increased His-Purkinje refractoriness in humans but had no effect on His-Purkinje conduction. The drug must be used judiciously in patients with a diseased His Purkinje system because it may cause atrioventricular block distal to His at fast heart rates. Sotalol had no effect on macrore-entry utilizing bundle branches. PMID- 10684395 TI - Investigative Concerns in Demonstrating Reduced Risk From Reversing Left Ventricular Hypertrophy. AB - BACKGROUND: To demonstrate reduced risk from reversing left ventricular hypertrophy (LVH) in hypertension, one must show that it is independent of blood pressure reduction. METHODS AND RESULTS: A feasibility study was conducted with 15 patients. The study employed 48-hour Holter recording, exercise treadmill (for ST-segment changes) and, as necessary, thallium scintigraphy and coronary angiography. All patients were treated for 3 months with quinapril (10 mg) and demonstrated decreased mean arterial pressure (125 +/- 3.1 vs 103 +/- 1.9 mmHg; P <.01) and left ventricular mass index (125 +/- 6.4 vs 104 +/- 4.9; P <.02) with preserved left ventricular function. There were no significant changes in these patients with moderate LVH in the incidence of arrhythmias; however, 4 of the 15 patients developed ST-segment changes prior to LVH reversal, and these changes did not recur in 3 patients following reversal of LVH or when pressure was allowed to rise. CONCLUSIONS: Ischemic changes, rather than development of arrhythmias, may be of greater value in demonstrating risk reduction with LVH reversal. Moreover, these preliminary data suggest pitfalls in demonstrating risk reduction after LVH reversal, indicating that more sensitive and adequate techniques are necessary to show risk reduction from LVH. PMID- 10684396 TI - A Randomized Multicenter Trial Comparing and Efficacy of Simvastatin and Fluvastatin. AB - BACKGROUND: Inhibitors of hydroxymethylglutaryl co-enzyme A reductase are widely used for the treatment of hypercholesterolemia. Physicians and third-party payers need an accurate measure of their relative potency and hypolipidemic efficacy. We have therefore compared simvastatin against fluvastatin, the newest member of this class. METHODS AND RESULTS: One hundred fifty-eight hypercholesterolemic patients in seven United States lipid clinics participated in this balanced double-blind incomplete block study. After a placebo-diet run-in period, patients received treatment with active drug for three consecutive 5-week periods, with measurement of lipids in a NHLBI-CDC standardized central laboratory at the end of each period. Each patient was randomly assigned to three of the following five treatments: simvastatin 5 mg, 10 mg, and 20 mg and fluvastatin 20 mg and 40 mg. The mean percent reductions in low density lipoprotein cholesterol from baseline were 21, 27, 32, 16, and 23 respectively. The simvastatin/fluvastatin milligram potency ratio was 6.8 (95% CI, 5.3-9.3). At the same 20 mg dose, simvastatin produced an effect on LDL cholesterol approximately double that of fluvastatin and resulted in 46% of patients achieving their National Cholesterol Education Program low density lipoprotein cholesterol target levels, compared to 12% for fluvastatin. CONCLUSIONS: Fluvastatin at its maximal dose of 40 mg daily is approximately equivalent to simvastatin 5 mg daily. Higher doses of simvastatin are considerably more effective in the treatment of primary hypercholesterolemia. PMID- 10684397 TI - Protective Effect of Elastase Inhibition Against Myocardial Dysfunction and Injury Induced by Ischemia and Reperfusion in Isolated Rat Hearts. AB - BACKGROUND: Elastase release has been incriminated in the genesis of reperfusion induced myocardial dysfunction and injury, and elastase inhibitors have been reported to reduce myocardial dysfunction in dogs subjected to coronary artery occlusion and reperfusion. METHODS AND RESULTS: To examine if elastase inhibition will modify myocardial dysfunction and injured induced by ischemia and reperfusion in isolated hearts, hearts from male Sprague Dawley rats were subjected to 30 minutes of total ischemia followed by 30 minutes of reperfusion. Ischemia-reperfusion resulted in myocardial dysfunction (increase in coronary perfusion pressure and decrease in myocardial contraction), injury (measured as creatine kinase release), and lipid peroxidation (myocardial malondialdehyde). Perfusion of hearts with an elastase inhibitor, ICI200,880, protected against myocardial dysfunction, injury and lipid peroxidation following ischemia reperfusion. As expected, perfusion with superoxide dismutase protected the hearts against hemodynamic deterioration following ischemia-reperfusion. In in vitro studies, there was no direct effect of ICI200,880 on superoxide anion generation. CONCLUSIONS: ICI200,880 seems to exert cardioprotective effects against ischemia-reperfusion-induced injury and myocardial dysfunction in isolated buffer-perfused hearts, most likely by an elastase-like protease inhibitory activity. PMID- 10684398 TI - Electrophysiologic Effects of Beta-blocking Agent, Tilisolol, on Isolated Guinea Pig Ventricular Myocytes. AB - BACKGROUND: Electrophysiologic effects of a beta-blocking agent, tilisolol, were studied with isolated guinea pig ventricular myocytes using the whole cell patch clamp technique. METHODS AND RESULTS: Tilisolol at 10 uM or higher concentrations prolonged action potential duration (APD) at 90% repolarization (APD(90)) and at 100 uM or higher concentrations shortened APD at 20% repolarization (APD(20)) without changes in resting membrane potential. At 10 uM concentration tilisolol prolonged APD(90) from 236.6 +/- 55.3 ms in the control to 253.4 +/- 52.4 ms (n = 16; P <.01), while APD(20) was unaffected. At 100 uM tilisolol, APD(20) was shortened from 143.6 +/- 15.7 ms in the control to 133.7 +/- 22.6 ms (n = 8; P <.05). Under voltage clamp, tilisolol decreased the delayed rectifier K(+) current (I(K1)). Applications of 10 uM and 100 uM tilisolol reduced the maximal conductance of I(K) by 35.7 +/- 3.5% and 47.4 +/- 3.5% of the control, respectively, without changes in voltage dependence (n = 10). Tilisolol at 100 uM decreased the L-type Ca(2+) current (I(Ca.L)) by 22.0 +/- 9.8% (n = 6) of the control, and the inactivation curve was shifted to a hyperpolarizing direction. CONCLUSIONS: Tilisolol has a direct membrane action to depress I(K) and I(Ca.L), in addition to its beta-receptor blocking action. PMID- 10684399 TI - Reduction of Myocardial Infarct Size in the Rabbit by a Carbohydrate Analog of Sialyl Lewis(x). AB - BACKGROUND: Available data suggest that the accumulation of neutrophils within the myocardium following an ischemic event plays an important role in the pathogenesis of myocardial ischemia/reperfusion injury. It is of interest, therefore, to develop pharmacologic agents designed to inhibit neutrophil adhesion to the endothelium. METHODS AND RESULTS: A synthetic carbohydrate analog to the P-selectin ligand sialyl Lewis(x) (sLe(x)) was evaluated for its ability to protect the myocardium from ischemia/reperfusion injury. Open chest anesthetized rabbits were subjected to 30 minutes occlusion of the left circumflex artery followed by 5 hours of reperfusion. Vehicle or sLe(x) analog (10 mg/kg) was administered intravenously before the onset of reperfusion and every hour during the reperfusion period. Myocardial infarct size in rabbits treated with the sLe(x) analog (10 mg/kg) was administered intravenously before the onset of reperfusion and every hour during the reperfusion period. Myocardial infarct size in rabbits treated with the sLe(x) analog was significantly reduced when compared to rabbits treated with vehicle (28 +/- 9% vs 57 +/- 10% of the area at risk, p <.05). The compound did not alter circulating neutrophil counts or myocardial oxygen demand as determined by the rate-pressure product. Furthermore, neutrophil accumulation within the ischemic region was decreased by 44% (P <.05) in the hearts of animals receiving sLe(x) analog as compared to vehicle. CONCLUSIONS: Carbohydrate derivatives of sLe(x) may be effective in reducing the degree of myocardial injury after ischemia/reperfusion. PMID- 10684400 TI - Hydrochlorothiazide Increases Efferent Glomerular Arteriolar Resistance in Spontaneously Hypertensive Rats. AB - BACKGROUND: Micropuncture studies were performed to determine the intrarenal hemodynamic effects of two conventional antihypertensive agents, hydrochlorothiazide (HCTZ) and hydralazine (HYDR) alone and in combination. METHODS AND RESULTS: Male spontaneously hypertensive and normotensive Wistar Kyoto rats (19 weeks old) were treated for 3 weeks with vehicle (control), HCTZ (80 mg/kg/d), HYDR (5 mg/kg/d), or combined therapy (HCTZ 30 mg/kg/d and HYDR 2 mg/kg/d). Each treatment significantly reduced arterial pressure while effective renal plasma flow, glomerular filtration rate and single nephron glomerular filtration rate were unaffected by any treatment in either strain. In spontaneously hypertensive rats HCTZ decreased single nephron plasma flow (111 +/ 8 to 84 +/- 4 nL/min; P <.05) but, despite this reduction, glomerular pressure remained unchanged (51.4 +/- 0.7 to 52.1 +/- 0.8 mmHg) attributable to increased efferent glomerular resistance (1.58 +/- 0.14 to 2.11 +/- 0.12 10 U; P <.05). By contrast, HYDR increased single nephron plasma flow (to 147 +/- 8 nL/min; P <.01) and decreased efferent glomerular resistance (to 1.09 +/- 0.09 U; P <.05). Combined treatment produced responses similar to HCTZ when used alone, thereby nullifying the beneficial efferent glomerular resistance effects: single nephron plasma flow +/- fell (to 89 +/- 7 nL/min; P <.05) and efferent glomerular resistance increased (to 2.05 +/- 0.17 U; P <.05). In Wistar Kyoto rats, HCTZ and combined treatment had no effect. HCTZ alone induced glomerular ischemia that was associated with efferent glomerular arteriolar constriction in these spontaneously hypertensive rats. CONCLUSIONS: These findings provide a possible explanation for the lack of improved renal target-organ damage in controlled multicenter trials employing thiazide diuretics. PMID- 10684401 TI - Differences in the Effects of d- and dl-Sotalol on Isolated Human Ventricular Muscle: Electromechanical Activity After Beta-Adrenoceptor Stimulation. AB - BACKGROUND: The racemate of sotalol (dl-sotalol) and its dextrorotatory isomer (d sotalol) are equally effective in increasing isolated cardiac action potential durations. dl-Sotalol, however, is reported to be more effective than d-sotalol in increasing ventricular effective refractoriness following coronary artery occlusion. These differences are attributed to the beta-adrenergic blocking properties of dl-sotalol. We wished to determine if in isolated human ventricular muscle preparations the effects of 30 uM d0 and dl-sotalol could be modified by preexposure to 1 uM isoproterenol. METHODS AND RESULTS: Microelectrodes were used to record action potential duration (APD) in the presence and absence of isoproterenol during continuous pacing. Preparations were obtained from explanted hears of transplant recipients suffering idiopathic cardiomyopathies. Without isoproterenol, APD measured at 90% of repolarization (APD(90)) was significantly increased by both d- and dl-sotalol (352.0 +/- 17.7 to 418.0 +/- 23.8 ms, P <.05; and 339.2 +/- 17.0 to 405.0 +/- 25.3 ms, P <.05; respectively). Isoproterenol alone, prior to sotalol exposure, tended to shorten APD(90) in the two groups first exposed to this beta-adenoceptor agonist and subsequently exposed to either d-sotalol or dl-sotalol (317.5 +/- 16.5 to 286.3 +/- 28.8 ms and 288.0 +/- 16.2 to 254.0 +/- 15.0 ms, respectively). dl-Sotalol significantly increased APD(90) from its baseline value after isoproterenol (288.0 +/- 16.2 to 359.0 +/- 25.1 ms, P <.005) while d-sotalol did not (317.5 +/- 16.5 to 316.2 +/- 28.5 ms, NS). CONCLUSIONS: The beta-adrenergic blocking properties of dl-sotalol may be important in determining antiarrhythmic efficacy when tonic sympathetic nervous activity is high. PMID- 10684402 TI - Amiodarone Therapy After Previous Sotalol-induced Torsade de Pointes: Analysis of AT Dispersion to Predict Proarrhythmia. AB - BACKGROUND: Polymorphic ventricular tachycardia of the torsade de pointes type represents, potentially, the most dangerous side effect of antiarrhythmic drugs that prolong ventricular repolarization. Much effort has been devoted to the identification of the degree of drug-associated QT prolongation that might predict the occurrence of torsade de pointes. However, there is still no general agreement as to which level of QT prolongation might be the harbinger of torsade and which may simply represent the manifestation of the class III antiarrhythmic effect of a given compound. METHODS AND RESULTS: A 70-year-old woman who had survived an episode of cardiac arrest outside of a hospital was treated with dl sotalol (320 mg/d). After 8 days of therapy, she developed two episodes of hemodynamically unstable torsade de pointes. Sotalol was withdrawn and after extensive diagnostic work, therapy with amiodarone therapy was comparable to that observed during sotalol exposure, the patient tolerated amiodarone and is now free of recurrent ventricular tachyarrhythmias over a follow-up period of 1 year. Analysis of QT dispersion in the surface electrocardiograph revealed a marked increase during sotalol therapy but not during amiodarone administration (77 vs 47 ms). During drug-free control, QT dispersion was 43 ms. CONCLUSIONS: These findings emphasize the potential usefulness of determination of QT dispersion from the surface ECG to assess disparity in ventricular recovery, which is known to favor the occurrence of torsade de pointes. These observations need to be corroborated in large prospective trials. Finally, this case report further emphasizes the low arrhythmogenic potential of amiodarone-an unexplained paradox, the understanding of which might provide insights for the development of newer antifibrillatory compounds. PMID- 10684403 TI - Data-driven Decisions: The Importance of Clinical Trials in Arrhythmia Management. AB - As a result of clinical trials, the measurement of arrhythmias has evolved over the past three decades. In the late 1960s, customary teaching was that ventricular premature depolarizations were dangerous and antiarrhythmic therapy, in hopes of reducing fatal consequences, became common place; however, following clinical trials such as CAST, IMPACT, and SWORD, we learned that, at least in postinfarct patients, arrhythmia suppression may lead to increased rather than reduced mortality. Such trials have led to a marked reduction in therapy of indiscriminate ventricular ectopy and have led to ongoing testing of specific subgroups identified as having particularly higher adverse prognostic risk. With the advent of cardiac monitoring and the confirmation that ventricular tachyarrhythmias are the most common cause for sudden death, their therapy, too, has evolved and matured, again aided by clinical trials. The ESVEM study prospectively examined the role of monitor-guided versus electrophysiologically guided drug therapy of ventricular tachyarrhythmias and confirmed that both approaches may have a role in reducing arrhythmic deaths-though the specific benefits of each technique remain somewhat unsettled. Both the ESVEM and CASCADE studies suggested that the most effective drugs for ventricular tachyarrhythmias are the class II/III drugs, sotalol and amiodarone, both appearing more effective than our older class I agents. These should now be viewed as the first-line drugs for these arrhythmias. The relative benefits of these two agents with respect to each other and to implantable cardioverter defibrillators, however, remains to be determined by further clinical trials, such as AVID and CIDS. The therapy of atrial tachyarrhythmias has similarly evolved with the aid of clinical observations. While rate control is required in all patients with atrial fibrillation, we have come to realize that the applications of antiarrhythmic drugs for the purpose of maintaining sinus rhythm must be used only selectively rather than uniformly. Both a meta-analysis by Coplen and colleagues and a report by the SPAF investigators suggested that with atrial arrhythmias, too, antiarrhythmic drug therapy may result in enhanced rather than reduced mortality in some circumstances. Additional clinical trials are needed to further elucidate the role of antiarrhythmic therapy of atrial fibrillation. PMID- 10684404 TI - Rise and Fall of Guided Antiarrhythmic Therapy for Ventricular Tachycardia and Fibrillation. PMID- 10684405 TI - The Effects of Amlodipine on Left Ventricular Mass and Diastolic Function in Concentric and Eccentric Left Ventricular Hypertrophy. AB - BACKGROUND: The effects of the antihypertensive therapy with amlodipine (5-10 mg/day) on left ventricular mass and diastolic function were examined in 30 mild to moderate essential hypertensive patients who have left ventricular hypertrophy (LVH) and diastolic dysfunction. METHODS AND RESULTS: Each patient's left ventricular mass was measured, and left ventricular diastolic function was assessed by echocardiographic Doppler examination at entry, and at 3 and 6 months after the initiation of the treatment. Amlodipine reduced both blood pressure (from 164 +/- 14/104 +/- 6 mmHg to 134 +/- 9/83 +/- 4 mmHg) and left ventricular mass index (from 160 +/- 30 g/m(2) to 137 +/- 26 g/m(2)) significantly at 3 months and both parameters maintained at these levels for 6 months. When the patients were classified according to the type of the LVH, a significant regression in left ventricular mass index was seen only in the patients who had concentric LVH was a relative wall thickness >/=0.44 (n = 16), but not in the eccentric LVH group (n = 14), although both groups were not significantly different from each other regarding the basal hemodynamic parameters, baseline left ventricular mass index and the decrease in blood pressure in response to amlodipine treatment. The mitral inflow E/A ratio did not show any significant change in either group. CONCLUSIONS: Amlodipine produced significant regression in LVH only in the patients with concentric LVH, but not those with eccentric LVH, while it did not change the diastolic dysfunction. Therefore, the type of LVH seems to be an important feature in determining the effects of antihypertensive treatment on left ventricular mass index. PMID- 10684406 TI - A Comparison of the Tolerability and Efficacy of Lovastatin 20 mg and Fluvastatin 20 mg in the Treatment of Primary Hypercholesterolemia. AB - BACKGROUND: To compare the cholesterol-lowering potency of fluvastatin and lovastatin, a randomized, prospective, open-label parallel study was conducted in patients eligible for drug therapy by National Cholesterol Education Program guidelines. The study was conducted at eight centers in the United States. METHODS AND RESULTS: Patients were required to follow a cholesterol-lowering diet and were withdrawn from all lipid-lowering agents for 4 seeks prior to study entry. Patients were randomized to receive lovastatin 20 mg of fluvastatin 20 mg daily for 6 weeks. The two treatment groups were comparable with respect to demographic and clinical characteristics. Baseline lipid levels in the two groups were comparable. Lovastatin was significantly more effective than fluvastatin in lowering total cholesterol (-;19.5% vs -12.8%, P <.001) and low-density lipoprotein cholesterol (-27.6% vs -18.2%, P <.001). Changes in high-density lipoprotein and triglyceride levels were comparable in the two groups. The differences in cholesterol lowering were similar in the three strata of coronary heart disease risk factor status as defined by the second NCEP Adult Treatment Panel. Both treatments were well tolerated. CONCLUSIONS: Across the three chronic heart disease risk strata, lovastatin appears to be significantly more potent than fluvastatin, on a per milligram basis, in lowering cholesterol levels. PMID- 10684407 TI - Efficacy and Tolerability of Low-dose Simvastatin and Niacin, Alone and in Combination, in Patients With Combined Hyperlipidemia: A Prospective Trial. AB - BACKGROUND: Combination lipid-lowering therapy may be desirable in patients with elevated low-density lipoprotein cholesterol, high triglycerides, and low high density lipoprotein cholesterol. This study was conducted to determine the lipid lowering efficacy of the combination of low-dose simvastatin and niacin in patients with combined hyperlipidemia and low high-density lipoprotein cholesterol. METHODS AND RESULTS: In this multicenter, prospective, randomized trial, 180 patients with hypercholesterolemia and hypertriglyceridemia and/or low high-density lipoprotein cholesterol were randomized to combination simvastatin (10 mg/day) and niacin (0.75 g/day) or to either drug alone for 9 weeks. The dose of niacin was doubled (from 0.75 g/day to 1.5 g/day) in both the combination and niacin arms for the remaining 8 weeks. The combination of simvastatin, 10 mg/day, and niacin, 1.5 g/day, reduced total, low-density lipoprotein, and very low density lipoprotein cholesterol and triglycerides by 24%, 29%, 45%, and 31%, respectively, while increasing high-density lipoprotein cholesterol by 31%. The addition of niacin to simvastatin did not enhance the low-density lipoprotein cholesterol by 31%. The addition of niacin to simvastatin did not enhance the low density lipoprotein cholesterol-lowering effect of simvastatin; however, the combination was more effective than either monotherapy at raising high-density lipoprotein cholesterol and lowering very low-density lipoprotein cholesterol (P <.05). More patients discontinued treatment because of an adverse event in the niacin (P <.03) and combination groups (P =.06) than the simvastatin group. CONCLUSIONS: Treatment of patients with combined hyperlipidemia and/or low high density lipoprotein with combination low-dose simvastatin and niacin resulted in large reductions in total, low-density lipoprotein, and very low-density lipoprotein cholesterol and increases in HDL cholesterol. Although the combination was well tolerated in the current trial, its safety needs to be evaluated in larger trials of longer duration. PMID- 10684408 TI - Atorvastatin, a New HMG-CoA Reductase Inhibitor as Monotherapy and Combined With Colestipol. AB - BACKGROUND: Atorvastatin, a new HMG-CoA reductase inhibitor in clinical development has demonstrated an acceptable safety profile and marked cholesterol and triglyceride reduction at doses ranging from 10-80 mg/day. Since bile acid sequestering resins are often used in combination with HMGRIs to enhance cholesterol reduction, this trial was conducted to explore the use of atorvastatin alone and combined with colestipol in patients with primary hyperlipidemia. METHODS AND RESULTS: One hundred six patients with low-density lipoprotein (LDL) cholesterol >4.1 mM/L (160 mg/dL) and plasma triglycerides <3.9 mM/L (350 mg/dL) were randomized to treatment consisting of 20 g/day colestipol, 10 mg/day atorvastatin, or 10 mg/day atorvastatin plus 20 g/day colestipol for 12 weeks. Percent change from baseline in lipid variables were measured. The atorvastatin group showed a significant reduction in LDL cholesterol of 35% after 12 weeks. Combination therapy provided an additional 10% reduction in LDL cholesterol over that observed for atorvastatin alone. Twenty-one percent of all patients in the atorvastatin monotherapy group experienced associated adverse events compared with 60% in the combination therapy group. Ninety percent of atorvastatin monotherapy patients were compliant at every visit compared with 75% receiving combination therapy. CONCLUSIONS: Although the combination of atorvastatin plus colestipol was more effective in lowering LDL cholesterol than atorvastatin alone, atorvastatin 10 mg/day monotherapy provided a better safety profile and improved patient compliance, which may result in improved long-term cholesterol control. PMID- 10684409 TI - Therapy of Sustained Ventricular Arrhythmias With Amiodarone: Prediction of Efficacy With Serial Electrophysiologic Studies. AB - BACKGROUND: Programmed electrical stimulation early during amiodarone therapy has poor prognostic capabilities; and persistent inducibility has been associated with a favorable outcome in a majority of patients. These observations result from studies that differed significantly in methodology. METHODS AND RESULTS: The authors prospectively enrolled 121 patients in a standardized amiodarone dosing protocol in which amiodarone was the only antiarrhythmic agent. Electrophysiologic testing was done after 2 and 6 weeks to determine noninducibility, predictive value, and the significance of drug-induced prolongation of tachycardia cycle length. The mean age of the patients in the study was 63.2 +/- 11.5 years, and their ejection fraction was 32.8 +/- 11.9%. Coronary artery disease was present in 103 (85%). At 2 weeks 17 patients (14%) were no longer inducible, whereas 104 patients (86%) remained inducible. Patients in these groups were similar in age and ejection fraction. During follow-up evaluation, recurrences (35% vs 24%; P =.44) and sudden death (12% vs 13.5%) were similar in the two groups. Thirty-five of 95 patients (32%) with sustained monomorphic ventricular tachycardia had more than 100 ms prolongation of their cycle length, which was hemodynamically well tolerated (partial response), but 60 did not (nonresponse). Patients with a partial response were older (66.5 vs 61.1 years; P =.02) and had longer QRS durations (143.2 vs 129.4 ms; P =.03). They also had increased recurrences (37% vs 17%; P =.01) and more sudden deaths (23% vs 8%; P =.02). At 6 weeks 11 of 76 patients studied were noninducible. They had a lower recurrence rate than those who remained inducible (8% vs 27%; P =.02) but a similar number of sudden deaths (8% vs 16%; P =.27). Thirty-two patients partially responded, and 31 patients did not respond. During follow-up examination these two groups had a similar number of recurrences (25% vs 29%; P =.76) and sudden deaths (16% vs 16%). CONCLUSIONS: Noninducibility at 2 or 6 weeks of amiodarone therapy did not identify patients at low risk of sudden death. In inducible patients, tachycardia cycle length prolongation, even when well tolerated, was not a marker for favorable outcome. Electrophysiologically guided therapy, therefore, offers little benefit over empiric amiodarone. PMID- 10684410 TI - Comparison of the Electromechanical Effects of Vesnarinone and Amrinone in Isolated Dog Purkinje Strands and Ventricular Trabeculae. AB - BACKGROUND: Conventional microelectrode techniques were used to compare the concentration-dependent effects of vesnarinone (0.1-100 uM) and amrinone (1 uM-1 mM) on action potential duration (APD) and developed force in both isolated dog ventricular trabeculae and Purkinje strands. METHODS AND RESULTS: Both drugs increased contractility of trabecular muscle preparations, while, in Purkinje strands, vesnarinone failed to increase developed force during continuous pacing at 2 Hz. Vesnarinone lengthened APD in both preparations; although this effect was more marked in Purkinje strands. Ventricular muscle APD was not affected by amrinone (1 uM to 1 mM), while, in Purkinje strands, amrinone produced a biphasic effect on APD. Low concentrations (1-100 uM) of amrinone shortened Purkinje fiber APD, while only the highest concentration (1 mM) used lengthened APD. In addition, in Purkinje strand preparations the effects of vesnarinone (10 uM) on APD and developed force were proportional to pacing cycle length at frequencies slower than 2 Hz; however, at frequencies faster than 2 Hz vesnarinone decreased developed force while APD was lengthened. In ventricular trabecular muscle preparations, the effects of vesnarinone were not affected by frequency. CONCLUSIONS: These results indicate clear differences between the effects of vesnarinone and amrinone in isolated cardiac preparations. These differences in experimental effects in isolated cardiac preparations may help provide an explanation for the disappointing clinical response of patients in heart failure to amrinone, while vesnarinone has appeared to be beneficial. PMID- 10684411 TI - Protective Effects of Ranolazine on Ventricular Fibrillation Induced by Activation of the ATP-Dependent Potassium Channel in the Rabbit Heart. AB - BACKGROUND: The authors studied the antifibrillatory effects of the adenosine triphosphate (ATP)-sparing metabolic modulator ranolazine in a rabbit isolated heart model in which ventricular fibrillation occurs under conditions of hypoxia/reoxygenation in the presence of the ATP-dependent potassium channel opener pinacidil. METHODS AND RESULTS: Ten minutes after ranolazine or vehicle administration, addition of pinacidil (1.25 uM) to the buffer was followed by a 12-minute hypoxic period and 40 minutes of reoxygenation. At a reduced concentration of ranolazine (10 uM), ventricular fibrillation occurred in 60% of the hearts, compared to 89% in the control group (P = NS). In contrast, only three of nine hearts (33%) treated with 20 uM ranolazine developed ventricular fibrillation (P <.05 vs vehicle). Hemodynamic parameters including coronary perfusion pressure, left ventricular developed pressure, and +/-dP/dt were not affected by the presence of ranolazine in the perfusion medium. Ranolazine did not prevent or modify the negative inotropic or coronary vasodilator actions of pinacidil, suggesting a mechanism of action independent of potassium channel antagonism. CONCLUSIONS: Ranolazine significantly reduced the incidence of ventricular fibrillation in the hypoxic/reoxygenated heart exposed to the ATP dependent potassium channel opener, pinacidil. The reported ability of ranolazine to prevent the decrease in cellular ATP during periods of a reduced oxygen supply may account for its observed antifibrillatory action. By maintaining intracellular ATP, ranolazine may modulate or prevent further opening of the ATP dependent potassium channel in response to hypoxia and/or pinacidil. PMID- 10684412 TI - Evidence of Na Current Contribution to the Transient Outward Current in Cardiac Ventricular Myocytes. AB - BACKGROUND: To study the transient outward current (I(to)) investigators often use sodium-free external solution to minimize the possible contamination of I(to) by sodium current. Removal of extracellular sodium creates reversal of sodium gradient and thus possibly contributing to I(to) mainly at positive potentials. METHODS AND RESULTS: To address this issue, whole-cell I(to) was recorded in sodium-free choline chloride and cobalt solutions, from rat ventricular myocytes known to exhibit a prominent I(to). Depolarizing pulse to 40 mV from -100 mV holding potential every 10 seconds elicited a fast activating and time-dependent inactivating components. The activation of I(to) was fast and complete within 10 ms at 40 mV, and the decay was rapid over the first 100 ms of the pulse and slower thereafter. External superfusion of the cell with 50 uM tetrodotoxin reversibly reduced I(to) amplitude by 25% from 1.47 +/- 0.2 to 1.1 +/- 0.3 nA (P <.04, n = 9). When sea anemone toxin (ATXII), known to selectively enhanced I(Na) by causing a delay in the inactivation gate, is applied to the cell, I(to) amplitude increased in a time- and dose-dependent manner (EC(50) =.86.4 nM). ATXII (100 nM) dramatically increased I(to) amplitude at all voltages between -20 and 60 mV (from 1.51 +/- 0.4 to 3.35 +/- 0.8 nA at 40 mV, P <.003, n = 12). Superfusion of cells with 5 mM 4-AP resulted in 82% reduction in I(to) amplitude at 40 mV (from 1.95 +/- 0.5 to 0.37 +/- 0.2 nA, P <.02, n = 8). Addition of ATXII to 4-AP containing solution increased peak I(to) by 965% (from 0.37+/-0.2 to 3.95 +/- 0.9, n = 8, P <.0003). However, in 11 other cells, addition of tetrodotoxin (50 uM) to the ATXII-containing solution blocked ATXII-induced outward current (from 3.51 +/- 0.64 nA to 1.60 +/- 0.17 nA, P <.05). The conductance (G(Ito)) was calculated by dividing peak I(to) by (Vm-E(K)), with an E(K) of -75 mV. G(Ito) was increased at all voltages (greater than -40 mV). Normalized G(Ito) was fitted by Boltzmann equation and ATXII did not significantly modify V(0.5) and k (from 20.5 +/- 3.9 to -17.0 +/- 3.5 mV for V(0.5), and 12.2 +/- 2.6 to 13.4 +/- 2.1 mV for k, n = 4). Also, atropine (1 uM) did not have any significant effect on I(to) (from 1.92 +/- 0.15 nA to 1.85 +/- 0.25 nA, n = 5). CONCLUSIONS: The results showed that, in sodium-free external solution I(to) is tetrodotoxin but not atropine sensitive. ATXII-induced I(to) increase is 4-aminopyridine insensitive but tetrodotoxin sensitive. These data suggest that outward Na current due to reversal of Na gradient in the absence of external Na contributes to I(to). Caution must be taken when studying kinetics and pharmacology of I(to) in external sodium-free solutions. PMID- 10684413 TI - Sudden Death During Flecainide Therapy for Atrial Fibrillation Complicating Wolff Parkinson-White Syndrome. AB - The Wolff-Parkinson-White syndrome can be complicated by atrial fibrillation that may increase morbidity and mortality. Different pharmacologic therapy, includes class IA, IC, and III agents, has been used in such cases with variable success. We now use less pharmacologic intervention with development of an electrode catheter ablation for accessory pathways. However, antiarrhythmic agents are still being used, especially when an electrode catheter ablation is unavailable or if a patient refuses such a procedure. Therefore, it is prudent that one understands each antiarrhythmic agents' electropharmacologic properties as well as its potential proarrhythmic effect in order to accurately assess each drug's risk-benefit ratio. We present a case that illustrates electropharmacologic properties of quinidine, flecainide, sotalol, and amiodarone on various cardiac tissues, as well as possible proarrhythmic effect of flecainide on a structurally normal heart. PMID- 10684414 TI - New Antithrombotic Drugs of Coronary Artery Disease. PMID- 10684415 TI - Prevention of Restenosis by Local Drug Delivery. AB - Local drug therapy for preventing restenosis after angioplasty has been investigated for over a decade. Biologically active agents ranging from drugs to genes can be delivered locally using a wide variety of catheters. Microspheres, liposomes, and polymers have been used to enhance drug retention at the delivery site. More recently stents have been investigated as devices to attain local drug delivery, either by coating with polymers, seeding with genetically modified cells or by using them as a source of local radiation. Though the best method of delivering agents locally remains undefined, this approach is likely to emerge as an essential mode of therapy in the near future. PMID- 10684416 TI - Influencing Mortality in Cardiac Disorders by Controlling Arrhythmias or by Cardioprotection: Whither Magnesium? PMID- 10684417 TI - Treatment Effect of Niaspan, a Controlled-release Niacin, in Patients With Hypercholesterolemia: A Placebo-controlled Trial. AB - BACKGROUND: The present study was designed to determine the efficacy and safety of Niaspan (Kos Pharmaceuticals, Inc, Hollywood, FL), a new controlled-release formulation of niacin, in the treatment of primary hyperlipidemia, the occurrence and severity of flushing events, and potential adverse effects, particularly hepatotoxicity. METHODS AND RESULTS: The study was conducted as a multicenter, randomized, double-blind, placebo-controlled, parallel comparison of Niaspan in doses of 1000 mg/day and 2000 mg/day, administered once a day at bedtime. One hundred twenty-two patients with low-density lipoprotein cholesterol levels >4.14 mM/L (160 mg/dL) with dietary intervention and high-density lipoprotein cholesterol 160 mg/dL were randomized to 4 weeks of treatment with one of the following regimens: cervastatin 0.1 mg twice daily, cerivastatin 0.2 mg once daily with the evening meal, cerivastatin 0.2 mg once daily at bedtime or placebo. All three active treatment groups produced statistically significant (P <.05) changes compared to aseline and placebo in total cholesterol (0.1 mg twice daily ?_18.9%; 0.2 mg once daily with the evening meal: ?_21.9%; 0.2 mg once daily at bedtime: ?_22.1%; placebo: 0.0%), low-density lipoprotein cholesterol (0.1 mg twice daily: ?_25.7%; 0.2 mg once daily with the evening meal: ?_29.4%; 0.2 mg once daily at bedtime: ?_30.4%; placebo: 1.4%) and high-density lipoprotein cholesterol (0.1 mg twice daily: 5.3%; 0.2 mg once daily with the evening meal: baseline and placebo, were also reduced by all active treatments (0.1 mg twice daily: ?_11.6% [P =.05]; 0.2 mg once daily with the evening meal: ?_11.6% [P =.05]; and 0.2 mg at bedtime: ?_10.9% [P =.07]). The percentage change in total cholesterol and low-density lipoprotein cholesterol after 4 weeks of therapy for the once-daily cerivastatin groups was statistically significantly greater (P <.05) than the cerivastatin twice daily regimen. A treatment responser was seen by 1 week of therapy and was maximal by 3 weeks. The drug was well tolerated in all three dosing regimens and resulted in no significant increase in biochemical or clinical side effects compared to placebo. CONCLUSION: Cerivastatin is a novel, highly potent, well-tolerated HMG-CoA reductase inhibitor that produces low-density lipoprotein cholesterol reductions of approximately 30% when administered at 0.2 mg once a day in the evenings. PMID- 10684438 TI - Long-term Treatment With Pravastatin Alone and in Combination With Gemfibrozil in Familial Type IIB Hyperlipoproteinemia or Combined Hyperlipidemia. AB - BACKGROUND: Pravastatin inhibits 3-hydroxy-3-methylglutaryl-coenzyme A reductase. It prevents mevalonate synthesis, reducing endogenous cholesterol production, and reduces cholesterol content in the liver, thus resulting in a down-regulation of low-density lipoprotein receptor production. Gemfibrozil reduces very low-density lipoprotein production and low-density lipoprotein-cholesterol level and increases very low-density lipoprotein catabolism. Therefore, it was suggested that combination therapy with both drugs could effect greater reduction of cholesterol levels as compared to pravastatin alone. The present study was carried out to evaluate the efficacy and safety of pravastatin as a monotherapy or in combination with gemfibrozil in the treatment of patients with familial type IIb hyperlipoproteinemia or familial combined hyperlipidemia. METHODS AND RESULTS: Forty-one patients were included in the study. All patients initially followed 6 weeks of hypolipidemic diet; subsequently they were randomized and received either 20 mg once daily of pravastatin alone (n = 13) or 20 mg of pravastatin together with 600 mg of gemfibrozil twice daily (n = 14). As a control, 14 patients were treated with diet only. The treatment lasted 24 months and clinical evaluation and laboratory tests were done at given time points. Both groups of treated patients showed an early reduction (3 months) of total (about 30% P <.01 vs controls), low-density lipoprotein (about 35%, P <.01 vs controls) and very low-density lipoprotein cholesterol levels (about 18%, P = NS). In contrast, high-density lipoprotein cholesterol levels increased significantly in patients treated with pravastatin and gemfibrozil (about 20%, P <.05 vs controls). Pravastatin treatment alone reduced the level of serum triglycerides as efficiently as in combination with gemfibrozil. Data showed a sustained normalization of lipid profile until 24 months. However, this effect was achieved in patients that had rather low levels of triglycerides. During the treatment we did not observe any difference in the incidence of possible drug-related side effects. Severe myopathy or rhabdomyolysis was not observed at the doses of the drugs used in our study. CONCLUSIONS: Therapy with pravastatin and in combination with gemfibrozil resulted in significant and sustained normalization of lipid profile in high-risk patients with familial type IIb hyperlipoproteinemia or familial combined hyperlipidemia. PMID- 10684439 TI - d-Sotalol Induces Marked Action Potential Prolongation and Early Afterdepolarizations in M but Not Empirical or Endocardial Cells of the Canine Ventricle. AB - BACKGROUND: Despite its class III antiarrhythmic actions, experimental and clinical studies have shown that d-sotalol can also be proarrhythmic; a recent clinical trial that evaluated d-sotalol in postmyocardial patients (SWORD) had to be prematurely interrupted because of the excess mortality in the treated group. Previous studies have demonstrated the existence of a marked heterogeneity across the ventricular wall; epicardial, endocardial, and M cells have been shown to display distinct electrophysiologic characteristics and pharmacologic behavior. The present study was designed to test the hypothesis that M cells are the primary target for the class III actions of d-sotalol in canine ventricular myocardium and may contribute to its proarrhythmic effects. METHODS AND RESULTS: We used standard microelectrode techniques to record transmembrane activity from endocardial, epicardial, midmyocardial, and transmural strips, isolated from the canine left ventricle. d-Sotalol (100 uM, 60 minutes of exposure, [K(+)]o = 4 mM) prolongs the action potential in the three cell types, but more so in M than epicardial or endocardial cells, especially at the slower rates. At a basic cycle length of 2000 ms, action potential duration after 90% repolarization increases from 199 +/- 20 to 247.5 +/- 28 ms in epicardium (n = 10), from 212 +/- 26 to 274 +/- 27 ms in endocardium (n = 11), and from 309 +/- 65 to 533 +/- 207 ms in M cells (n = 13). d-Sotalol produces a marked steepening of action potential duration-rate relationships of M cells and an upward shift of restitution of action potential duration curves, more accentuated in M cells. Early afterdepolarizations were observed at slow rates (basic cycle lengths > 1000 ms) in 7 of 13 M cell preparation s(54%) but not in endocardial or epicardial preparations. A sudden acceleration of the rate could also induce a transient prolongation of the action potential and early afterdepolarization activity. CONCLUSION: In canine ventricular tissues, d-sotalol manifests its class III effects preferentially in the M cells, leading to the development of early afterdepolarizations and a marked increase in transmural dispersion of repolarization. The data suggest an important role of M cells in the proarrhythmic effects of the drug. PMID- 10684440 TI - The Novel Class III Antiarrhythmic Agent MS-551 Blocks the Cardiac Inward Rectifier With Greater Potency Than Sotalol or E-4031: Possible Relevance to Reverse Use Dependence. AB - BACKGROUND: The tendency for the electrophysiologic effect of class III antiarrhythmic agents (action potential prolongation) to be diminished at faster heart rates represents a major drawback of this class of drug and is usually referred to as "reverse use dependence." A novel class III agent, MS-551, has recently been reported to exhibit less reverse use dependence than E-4031. We set out to investigate whether this observation may be due to differential blockade of the inward rectifier current (i(K1)) by these drugs. METHODS AND RESULTS: We recorded i(K1) using single channel methods and cell attached patch configurations, with standard patch clamp technology. Neither E-4031 nor racemic sotalol in concentrations up to 100 uM had any significant effect on the open probability or kinetics of i(K1) without altering the single-channel conductance. Openings to subconductance levels were abolished in three of six patches in which they had been frequently present in the absence of drug. MS-551 had no effect on mean channel open time but increased the slower component of the closed time. CONCLUSIONS: MS-551, unlike E-4031 and sotalol, appears to produce significant blockade of the inwardly rectifying potassium channel at clinically relevant concentrations. We propose that this might provide a partial explanation for the observed differences in their response to rate changes. PMID- 10684441 TI - Estradiol, Administered Acutely, Protects Ischemic Myocardium in Both Female and Male Rabbits. AB - BACKGROUND: The benefits of chronic administration of estrogen to postmenopausal women are well documented; however, the acute effects of exogenous estradiol on myocardium after coronary artery occlusion and reperfusion in male and female animal models are unknown. We tested the influence of acute pretreatment with estradiol on the development of myocardial necrosis in two protocols, studying intact anesthetized female and male rabbits. METHODS AND RESULTS: 17beta estradiol (1 mg) was given 15 minutes before coronary artery occlusion in the treated groups (n = 10 females, 10 males); control rabbits (n = 11 females, 10 males) received water. All rabbits underwent 30 minutes of coronary artery occlusion and 4 hours of reperfusion. Myocardial blood flow was similar between groups at 10 minutes after treatment and during coronary artery occlusion and reperfusion. Thus estradiol did not increase blood flow. Heart rate and systemic pressure were also similar between groups. Estradiol levels during coronary artery occlusion were 1-8 pg/mL in untreated female and male rabbits and 66 +/- 28 (male) and 352 +/- 273 (female) in treated rabbits. Although the size of the ischemic risk zones was similar in both groups in both protocols, estradiol treated rabbits of both sexes developed significantly less necrosis. Infarct size as a percent of the risk region was 10 +/- 1% in female estradiol-treated rabbits compared with 23 +/- 5% in controls (P <.03) and 16 +/- 4% in estradiol-treated male rabbits compared with 31 +/- 5% in control males (P =.03). Although male rabbits had larger infarcts than female rabbits, sex was not a significant covariate for infarct size. CONCLUSIONS: Estradiol exerts a protective effect on ischemic myocardium that is not associated with an increase in myocardial blood flow or alteration in hemodynamics. This study shows that acute administration of estrogen before coronary artery occlusion reduces infarct size in both male and female rabbits. PMID- 10684442 TI - Role of Oxidative Stress in Amiodarone-induced Toxicity. AB - BACKGROUND: The clinical usefulness of amiodarone for the treatment of cardiac arrhythmias is limited by multiorgan toxicity, especially pulmonary and hepatic. There are conflicting reports in the literature regarding the role of free radicals in the initiation of amiodarone-induced toxicity. We evaluated the possible oxidative stress in a chronic model that is known to manifest pulmonary toxicity. METHODS AND RESULTS: A group of 20 Fischer-344 rats were injected with 60 mg/kg/day of amiodarone for 21 days. A control group of 20 animals received only saline injections. The alveolar macrophages obtained by lung lavage were incubated with hydroethidine and opsonized green fluorescent zymosan particles to measure oxidative and phagocytic activities by flow cytometry. Malondialdehyde levels were measured to assess the extent of lipid peroxidation in lung, liver, spleen, kidney, and heart. Total phospholipid levels in all the collected tissues and distribution of phospholipid classes in the lung and the liver were measured. The levels of amiodarone and its metabolite desethylamiodarone in serum and all collected tissues were measured by high-performance liquid chromatography. The phagocytic activity of th emacrophages from treated animals was decreased by 18 22% (P <.03) compared to controls; however, the oxidative activities of control and treated groups were not significantly different. The tissue malondialdehyde levels were not significantly different except in the spleen where they increased after amiodarone treatment (18.2 +/- 1.1 vs 23.7 +/- 2.8 uM/g tissue, P <.0001). Malondyaldehyde levels were not significantly different when normalized to lipid phosphorous content. Lung, liver, and spleen showed significantly higher phospholipid levels in the treated group. The tissue amiodarone and desethylamiodarone levels in the treated group were highest in spleen followed by lung, liver, kidney, and heart. CONCLUSIONS: The results show that amiodarone induced pulmonary and hepatic toxicity is not directly mediated by oxidative stress; however, increased lipid peroxidation in the spleen, although secondary to phospholipidosis, may be physiologically significant. PMID- 10684443 TI - Electrophysiology of Myocardial Cells in the Epicardial, Midmyocardial, and Endocardial Layers of the Ventricle. AB - The recent discovery of multiple myocardial cell types in the ventricular wall of most species has prompted a reevaluation of several electrophysiologic and electrocardiographic findings. This review briefly presents the salient electrophysiologic features of myocardial cells in the epicardial, midmyocardial and endocardial regions of the ventricle. The epicardial action potential exhibits a prominent notch between phase 1 and phase 2 that results in a spike and dome configuration. The notch is smaller in midmyocardial cells and absent in endocardial cells. The action potential notch is due to the presence of a transient outward current (I(to)), which diminishes in amplitude from the epicardial to endocardial surfaces. Midmyocardial or "M cells" exhibit electrophysiologic features intermediate between those of myocardial and conducting cells. M cells differ from epicardial and endocardial cells primarily in their response to slowing of the stimulation rate. These cells display an exaggerated prolongation of action potential duration at moderate to slow rates of stimulation. The atypical response in M cells reflects decreased levels of the delayed rectifier K(+) current (I(K)) in this cell type (dV/dt) compared to epicardial or endocardial cells. These electrophysiologic distinctions contribute to differences in the responsiveness of the various cell types to pharmacologic agents and disease. Also, the dispersion of repolarization created between epicardium and endocardium in the early phases of the action potential, and between M cells and other ventricular layers during late repolarization, may explain the J wave and U wave of the electrocardiogram, respectively. PMID- 10684444 TI - The M Cell. PMID- 10684445 TI - Efficacy and Safety of Nifedipine Coat-Core versus Amlodipine in Patients With Mild to Moderate Essential Hypertension: Comparison of 24-Hour Mean Ambulatory Diastolic Blood Pressure. AB - BACKGROUND: Calcium channel blockers have been successfully used for the treatment of hypertension. In this study, the antihypertensive efficacy and safety of the dihydropyridine calcium channel blockers nifedipine coat-core 30 mg and amlodipine 5 mg were evaluated. METHODS: This multicenter, double-blind, prospective, randomized, parallel-arm study compared once daily administration of nifedipine coat-core 30 mg with once daily amlodipine 5 mg in subjects with mild to-moderate essential hypertension. A 4-week placebo run-in period was followed by an 8-week active treatment period. Blood pressure reduction was measured by ambulatory blood pressure monitoring and casual office blood pressure measured by mercury sphygmomanometer. RESULTS: Nifedipine coat-core and amlodipine produced equivalent reductions in mean diastolic blood pressure, as determined by 24-hour ambulatory blood pressure monitoring. Mean reduction in diastolic blood pressure was 5.4 mmHg and 5.8 mmHg for nifedipine coat-core and amlodipine, respectively. Both drugs were well tolerated and neither treatment resulted in a significant change in heart rate. CONCLUSIONS: Nifedipine coat-core 30 mg once-daily is comparable to amlodipine 5 mg once-daily for blood pressure reduction. PMID- 10684446 TI - Urinary NItrotyrosine Content as a Marker of Peroxynitrite-induced Tolerance to Organic NItrates. AB - BACKGROUND: Anti-ischemic therapy with nitrovaasodilators as NO-donors is complicated by the induction of tolerance. When nitrovasodilators are metabolized to release NO there is a considerable coproduction of oxygen-derived radicals leading to a diminished cyclic GMP production and to impaired vasomotory responses. We analyzed in vivo the glyceroltrinitrate-induced generation of strong oxidative/nitrating compounds contributing to development of tolerance. METHODS AND RESULTS: In 16 patients we studied the urinary nitrotyrosine excretion during either (1) placebo control conditions, (2) 2-day nonintermittent transdermal nitroglycerin administration (0.4 mg/h), (3) 2-day nonintermittent glyceroltrinitrate administration (0.4 mg/h) along with a continuous infusion of vitamin C (55 ug/kg/min) as an antioxidant, or (4) with vitamin C but without glyceroltrinitrate (diminished urinary nitrotyrosine content of 34 +/- 18 ug/day observed). Glyceroltrinitrate administration augmented urinary nitrotyrosine from 56 +/- 24 (basal) to 186 +/- 32 ug/day (glyceroltrinitrate tolerance). Coadministration of vitamin C caused complete elimination of tolerance and a decrease in urinary nitrotyrosine to 130 +/- 28 ug/day. Glyceroltrinitrate induced formation of oxidants was confirmed in vitro comparing glyceroltrinitrate induced and peroxynitrite-induced tachyphylaxis in isolated perfused rabbit hearts and analyzing tolerance-induced inactivation of solbule guanylyl cyclase in cultured aortic smooth muscle cells. CONCLUSIONS: Augmented urinary nitrotyrosine excretion during glyceroltrinitrate administration reflects enhanced formation of peroxynitrite and of nitrotyrosine. Glyceroltrinitrate induced tolerance is the result of oxidative stress and can be suppressed by additional antioxidant therapy aimed to prevent glyceroltrinitrate-induced formation and/or actions of peroxynitrite. PMID- 10684447 TI - Digoxin's Minimal Inotropic Effect Is Not Limited by Sodium-Calcium Exchange in the Intact Immature Rabbit Heart. AB - BACKGROUND: In the intact immature heart, how much digoxin can drive sodium calcium exchange has not been studied in the context of sodium-calcium exchanger abundance. METHODS AND RESULTS: The effects of digoxin and low potassium on contractility in the intact, paced and isovolumically contracting immature rabbit heart were studied in both the absence and presence of L-type calcium channel blockade. Without calcium channel blockade, digoxin increased contractility minimally and only at 10(_6) M/L. In contrast, low potassium (2.2 mM/L) substantially increased contractility in all experiments, a result indicating abundant sodium-calcium exchanger activity. During nifedipine-induced calcium channel blockade, digoxin (10(_6) M/L) allowed modest recovery of contractility, whereas digoxin and low potassium together allowed complete recovery as assessed by dP/dt(max); however, all hearts so perfused subsequently developed ventricular fibrillation, presumably because of calcium overload. CONCLUSIONS: In intact immature rabbit heart, digoxin can drive sodium-calcium exchange and thus increase contractility to only a minimal extent. This effect does not appear to be limited by intrinsic exchanger activity, which appears abundant in this preparation. Rather, digoxin's inability to drive the sodium-calcium exchanger may be due to developmental differences in binding to the sodium pump. The sodium calcium exchanger itself seems capable not only of providing enough intracellular calcium for normal contraction, but also of overloading the myocardium with calcium, despite L-type calcium channel blockade. PMID- 10684448 TI - Nitric Oxide Synthesis Inhibition and Role of P-selectin in Leukocyte Adhesion to Vascular Tissues. AB - BACKGROUND: This study was designed to examine the role of P-selectin expression in leukocyte adhesion to endothelium caused by inhibition of nitric oxide synthesis. METHODS AND RESULTS: Rat aortic rings were treated with the nitric oxide synthesis inhibitor N(omicron)-nitro-l-arginine methyl ester (l-NAME) for 2 hours. Parallel sets of aortic rings were pretreated with the nitric oxide precursor l-arginine or posttreated with a specific monoclonal antibody against P selectin. Some rings were used for determination of vasoreactivity in response to norepinephrine and acetylcholine, while other rings were incubated with autologous unlabeled leukocytes or Biotin-FITC labeled leukocytes for 30 minutes. Leukocyte adhesion to vascular endothelium was determined by scanning electron microscopy. l-NAME enhanced the contractile response in response to norepinephrine, suppressed the relaxant response to acetyleholine, promoted leukocyte adherence to the endothelium and resulted in P-selectin expression on the aortic endothelium. Pretreatment of aortic rings with l-arginine reversed the l-NAME-mediated changes in vasoreactivity in response to norepinephrine and acetyleholine and attenuated the l-NAME-enhanced leukocyte adhesion to endothelial intima. P-selectin treatment, on the other hand, had no effect on l NAME-mediated changes. Intraperitoneal administration of l-NAME resulted in a significant decrease in plasma nitrite level, a small, but significant, increase in lung and spleen myeloperoxidase activity, and a significant increase in leukocyte deposition in lung and spleen. The l-NAME-mediated increase in myeloperoxidase activity and leukocyte deposition in the spleen, but not in the lungs, was abolished by treatment of rats with the P-selectin antagonist CY1503 administered 30 minutes prior to l-NAME. CONCLUSIONS: These observations indicate that a reduction in nitric oxide synthesis enhances leukocyte adhesion to aortic endothelium and in visceral tissues. While P-selectin expression is evident in some of the experimental models of leukocyte adhesion to endothelium under conditions of nitric oxide inhibition, the role of P-selectin expression remains unclear. PMID- 10684449 TI - Alternations in beta-Adrenoceptor Mechanisms in Hearts Perfused With Xanthine Plus Xanthine Oxidase. AB - BACKGROUND: Although beta-adrenoceptors and adenylyl cyclase are known to be affected upon exposing cardiac membranes to some oxyradical generating systems, the results are conflicting. Furthermore, functional significance of alterations in the beta-adrenoceptor-adenylyl cyclase systems in terms of changes in the inotropic responses to catecholamines is not clear. METHODS AND RESULTS: The positive inotropic effect of isoproterenol was augmented on perfusing the isolated rat hearts with xanthine (X) plus xanthine oxidase (XO) for 5 minutes but was attenuated by perfusion for 15 minutes. The isoproterenol-stimulated adenylyl cyclase activity in cardiac membranes showed an increase at 10 minutes and a decrease at 30 minutes perfusion of hearts with X plus XO. The density of beta-adrenoceptors in cardiac membraners was reduced after 10 minutes and 30 minutes of perfusion with X plus XO, whereas the affinity of beta-adrenoceptors was increased after 10 minutes and reduced after 30 minutes. Although beta adrenoceptors was increased after 10 minutes and reduced after 30 minutes. Although beta-adrenoceptors were unaltered by 10 minutes of perfusion with X plus XO, their affinity was increased and density was decreased by 30 minutes of perfusion. The agonist competition curves using isoproterenol indicated an increase in the number of coupled receptors in the high affinity state on 10 minutes of perfusion and an increase in the low affinity state of coupled receptor due to 30 minutes of perfusion with X plus XO. The basal as well as forskolin-, NaF- and Gpp(NH)p-stimulated adenylyl cyclase activities in cardiac membranes exhibited an increase after 10 minutes and decrease after 30 minutes of perfusion with X plus XO. Although the presence of superoxide dismutase plus catalase in the perfusion medium prevented most of the alterations due to X plus XO, it did not alter the increased affinity of the beta-adrenoceptor upon perfusing hearts for 10 minutes with X plus XO. CONCLUSIONS: The results in this study suggest the biphasic nature of the oxyradical-induced alterations in both the inotropic responses to catecholamines and the beta-adrenoceptor-mediated signal transduction mechanism in the heart. PMID- 10684450 TI - Antiarrhythmic and Arrhythmogenic Actions of Varying Levels of Extracellular Magnesium: Possible Cellular Basis for the Differences in the Efficacy of magnesium and Lidocaine in Torsade de Pointes. AB - BACKGROUND: In recent years, there has been an increasing use of antiarrhythmic drugs that act predominantly by prolonging myocardial repolarization. An inevitable electrophysiologic consequence of these drugs is the development of torsade de pointes as a proarrhythmic reaction. Both intravenous lidocaine and magnesium sulphate have been used in the acute control of such a proarrhythmia. Their electrophysiologic mechanisms in this setting are not well defined. METHODS AND RESULTS: Using the standard microelectrode techniques, the effects of magnesium (Mg) and lidocaine on action potential duration (APD), and on barium induced spontaneous action potentials, were studied in canine Purkinje fiber preparations. The objective was to clarify the direct and indirect effects of magnesium on triggered activities due to early afterdepolarizations. Superfusion in media with 0.1 mM Mg and 2.5 mM K produced more pronounced increases in APD measured at -20mV repolarization time [APD(20)] than those in a solution with 5 mM K. This effect was further enhanced at lower stimulation frequencies. The striking prolongation of APD(20) by solutions with low potassium concentrations diminished as the Mg concentration was increased. In solutions with 2.5 mM K, Mg produced concentration-dependent decreases in APD(20). This effect was greater at lower stimulation frequencies. Lidocaine at 4.0 x 10(_5) M produced a marked shortening of the APD in the entire firing frequency of the abnormal automaticity in a concentration-dependent manner. With 10 mM Mg, such action potentials appeared only sporadically. Magnesium also decreased the amplitude and the maximum upstroke velocity of these action potentials. In contrast, lidocaine at 4.0 x 10(-5) M exhibited no significant effects on action potentials due to barium-induced abnormal automaticity, or on additional depolarizations developing from the repolarization phase of these action potentials. CONCLUSIONS: The data indicate that (i) hypomagnesemia may be arrhythmogenic when combined with hypokalemia and bradycardia leading to a prolongation of the plateau phase of the action potential, (ii) magnesium administration may suppress triggered activities mainly by a direct inhibition of the development of triggered potentials, and (iii) lidocaine may suppress triggered potentials only indirectly by preventing the development of early afterdepolarizations due to the shortening effect on the APD. These findings are consistent with the clinical observation of a high incidence of torsade de pointes in the setting of hypokalemia and hypomagnesemia introduced by a chronic diuretic therapy. They are also consistent with the marked effectiveness of intravenous Mg relative to the inconsistent clinical effects of lidocaine in controlling torsade de pointes. PMID- 10684451 TI - Restoring Sinus Rhythm in Patients With Atrial Flutter and Fibrillation: Pharmacologic or Electrical Cardioversion? AB - Atrial fibrillation and atrial flutter, the most frequently encountered tachyarrhythmias requiring treatment, have become a major focus for clinical and basic research in recent years. Restoration and maintenance of sinus rhythmn, having been shown to improve exercise capacity, alleviate symptoms, and reduce the incidence of thromboembolic events, may be the optimal management strategy. Identification of the safest, most efficacious and cost-effective means of restoring sinus rhythm is necessary prior to the institution of optimal antiarrhythmic therapy to maintain sinus rhythm. Potential advantages of pharmacologic compared with electrical cardioversion include lack of need for general anesthesia and likely lower cost. Pharmacologic conversion include lack of need for general anesthesia and likely lower cost. Pharmacologic conversion has been accomplished with drugs that prolong atrial refractorinerss, including class Ia (quinidine, procainamide, disopyramide), class Ic (flecainide, propafenone), and class II (sotalol, amiodarone) compounds. The so-called pure class III agents were created to overcome the blocker side effects of sotalol and the complex pharmacodynamic profile of amiodarone. Two such agents are dofetilide, which selectively blocks the rapid component of the delayed rectifier current (Ikr) and ibutilide, which augments the slow inward sodium current, with a smaller component of action mediated by the block of Ikr. Reported overall conversion rates for recent onset atrial fibrillation and atrial flutter were 31% and 54% for difetilide, respectively, and 29-31% and 38-63%, respectively, for ibutilide. Proarrhythmia, manifested as polymorphic ventricular tachycardia requiring cardioversion, was a significant early side effect of both agents. Data from clinical trtials with these new agents, combined with increasing nowledge of the electrophysiologic substrate for these arrhythmias, has renewed initerest in the development of safer, more efficacious class IIIdrugs for atrial fibrillation and atrial flutter conversion. PMID- 10684453 TI - Reflections on Recent Clinical Trials in Patients With Heart Failure and Those With Reduced Ventricular Function. PMID- 10684452 TI - Increase in AT1 Receptors as a Mechanism of ACE Inhibition-induced Cough. PMID- 10684454 TI - Effects of Phentolamine on Responses to PAMP in the Hindquarters Vascular Bed of the Rat. AB - BACKGROUND: Responses to proadrenomedullin NH(2)-terminal 20 peptide (PAMP), a novel hypotensive peptide formed from preproadrenomedullin, and the effects of inhibition of adrenergic vasomotor tone with the alpha-receptor antagonist, phenolamine, on responses to PAMP were investigated in the systemic and hindquarters vascular bed of the rat. METHODS AND RESULTS: Intravenous injections of PAMP decreased systemic arterial pressure and in the hindquarters vascular bed of the rat under conditions of controlled hindquarters blood flow, intra-arterial injections of PAMP decreased perfusion pressure in a dose-related manner. Following administration of the alpha receptor blocking agent, phenotlamine, systemic depressor and hindquarters vasodilator responses to PAMP were not significantly decreased, whereas phentolamine significantly decreased systemic and hindquarters pressor responses to norepinephrine. Phentolamine had no significant effect on vasodilator responses to bradykinin, albuterol, or to nitroglycerin. CONCLUSIONS: The present data show that PAMP has significant systemic vasodepressor and vasodilator activity in the hindquarters vascular bed of the rat and suggest that vasodepressor and hindquarters vasodilator responses to PAMP are not dependent upon the presence of adrenergic vasomotor tone. PMID- 10684455 TI - Differential Effects of Protein Kinase C Activators and Inhibitors on alpha- and beta-Adrenoceptor-mediated Positive Inotropic Effect in Isolated Rabbit Papillary Muscle. AB - BACKGROUND: A number of novel agents that activate or inhibit protein kinase C (PKC) in vitro have been developed to evaluate the physiologic role of PKC in regulation of cellular function. However, most of the PKC inhibitors also affect the protein kinase A, and the effects of these agents in intact myocardium remain still controversial. The present study was carried out to examine the effects of these agents on the positive inotropic effect (PIE) medicated by alpha- and beta adrenoceptors in isolated rabbit papillary muscle. METHODS AND RESULTS: A potent PKC activator, phorbol 12, 13-dibutyrate (PDBu) at 10 and 30 nM, induced a significant PIE. PDBu at 3 nM and higher inhibited the alpha-mediated PIE and abolished it at 100 nM without affecting the beta-mediated PIE. Phorbol 12 myrisate 13-acetate (PMA) and 1-oleyl-2-acetyl-sn-glycerol (OAG) elicited a similar selective inhibitory action on the alpha-mediated PIE. The PIE of PDBu was abolished by chelerythrine and partially inhibited by staurosporine, but H-7 or calphostin-C did not affect the PIE. These PKC inhibitors consistently inhibited the alpha-mediated PIE by 20-30% at concentrations that they did not affect the beta-mediated PIE. None of the PKC inhibitors influence the PDBu induced inhibitory action on the alpha-mediated PIE, an indication that they failed to reach the site of the inhibitory action of PDBu. CONCLUSION: Selective modulation by the PKC activators and inhibitors of the alpha-mediated PIE with little effect on the beta-mediated PIE implies that the activation of PKC has a physiological relevance to the alpha-mediated PIE. However, the externally administered PKC activators do not mimic the effect of diacylglycerol that is generated endogenously by alpha-stimulation. By contrast, externally applied PKC inhibitors selectively antagonize the alpha-adrenoreceptor-mediated PIE in rabbit ventricular myocardium. PMID- 10684456 TI - Pinacidil's Effects on Defibrillation Outcomes: Role of Increased Potassium Conductance Via the K(ATP) Channel. AB - BACKGROUND: It has been shown that the inhibition of potassium ion conductance decreases defibrillation threshold. We postulated that if potassium conductance is a primary mechanism affecting defibrillation threshold values, then increasing potassium ion conductance will increase defibrillation values. The primary objective of this study was to determine if the ATP-dependent potassium (K(ATP)) channel opener pinacidil would increase defibrillation threshold values. The second objective was to prove that the observed changes were due to potassium conductance by using the K(ATP) inhibitor, glyburide, to reverse the electrophysiologic actions of pinacidil. The third objective was to determine if the electrophysiology action sof pinacidil correlate with changes in defibrillation threshold value. METHODS AND RESULTS: Domestic farm swine (n = 14) were anesthetized and intubated. Subsequently, they were instrumented with monophasic action potential catheters and epicardial defibrillation patches. Defibrillation threshold values, action potential duration, effective refractory period, and ventricular fibrillation cycle length were determined at baseline and during treatment phase 1 and treatment phase 2. Pigs were randomized into 2 groups: group 1 (n = 6) received D(5)W in treatment phase one followed by D(5)W in treatment phase 2 and group 2 (n = 8) received pinacidil in treatment phase one followed by the addition of glyburide in treatment phase two. DFT(ED50) did not change at baseline, treatment phase one or treatment phase two for group 1 (10.5 +/- 2, 11.1 +/- 1.7, 10.5 +/- 1.0 J) or for group 2 (10.1 +/- 2.2, 11.4 +/- 4.2, 11.4 +/- 3.0 J). Electrophysiologic parameters )QRS, effective refractory period, action potential duration(90), and ventricular fibrillation cycle length) were not significantly changed from baseline in group 1. In contrast, effective refractory period, action potential duration(90), and ventricular fibrillation cycle length significantly decreased at all recorded sites after the administration of pinacidil in group 2 (range of 7-13%, 6-9%, and 12-17%, respectively). However, pinacidil did not change the basal level of dispersion in effective refractory period, action potential duration, and ventricular fibrillation cycle length during paced rhythm or ventricular fibrillation. Glyburide reversed pinacidil's electrophysiologic actions. CONCLUSIONS: Pinacidil does not alter defibrillation threshold, but it reduces effective refractory period, action potential duration, and ventricular fibrillation cycle length and does not increase electrical heterogeneity. Therefore, changes in potassium channel conductance as well as shortening repolarization are unlikely primary mechanisms for elevating defibrillation threshold. PMID- 10684457 TI - Local Delivery of an Ultra-short-acting Nitric Oxide-releasing Compound, DMHD/NO, Is Highly Effective in Inhibiting Acute Platelet-Thrombus Formation on Injured Arterial Strips. AB - BACKGROUND: Nitric oxide (NO) plays an important role in modulating platelet vessel wall interaction following vascular injury. We exampled the effects of local infusion of an ultra-short-acting NO-releasing compound: NO adduct of N, N' dimethylhexanediamine (DMHD/NO), sodium nitroprusside, intravenous nitroglycerin, and aspirin on acute platelet-thrombus formation under conditions of high-shear blood flow in a rabbit extracorporeal perfusion model. MATERIALS AND METHODS: Strips of porcine aortic media were perfused in a Badimon chamber with arterial blood from 20 New Zealand White rabbits for 10 minutes at a shear rate of 1700 s( 1). Thrombus formation was quantified by morphometric analysis of thrombus area. Effects on collagen-induced platelet aggregation, blood pressure, bleeding time, and activated clotting time were also examined. RESULTS: DMHD/NO inhibited thrombus area and platelet aggregation in a dose-dependent manner with a 90% reduction in thrombus area (0.018 +/- 0.039 vs 0.215 +/- 0.085 mm(2)/mm control, P <.001) and a 50% reduction in platelet aggregation (4.8 +/- 4.4 vs 9.9 +/- 4.1 Omicron control, P =.04) at the highest dose of 1.0 nM/kg and 100 uM/L, respectively, without any effects on blood pressure, bleeding time, or activated clotting time. In contrast, equimolar concentrations of sodium nitroprusside and intravenous nitroglycerin had significantly reduced effects on thrombus area compared to DMHD/NO and were associated with significant reductions in blood pressure and prolongation of bleeding time. Aspirin had no effect on thrombus area at 1 uM/kg but reduced thrombus area and prolonged bleeding time at 2 and 5 uM/kg. CONCLUSIONS: Local delivery of DMHD/NO produced a 90% inhibition of experimental acute platelet-thrombosis under high-shear flow conditions without producing adverse systemic hemodynamic or hemostatic effects. Thus, inhibition of thrombus formation by local delivery of a rapidly acting NO donor may be an effective strategy for prevention of arterial injury-induced thrombosis. PMID- 10684458 TI - Electrophysiologic Effects of the New Class III Antiarrhythmic Drug Dofetilide in an Experimental Canine Model of Pacing-induced Atrial Fibrillation. AB - BACKGROUND: Dofetilide is a new class III antiarrhythmic drug currently under investigation for the treatment of supraventricular arrhythmias in humans. Dofetilide have been previously shown to be highly effective in terminating and suppressing reentrant atrial flutter in the experimental canine crush-injury model, in which its antiarrhythmic efficacy was correlated with prolongation of wavelength and reduction in dispersion of refractoriness, effects not produced by the class IA antiarrhythmic drug quinidine. The purpose of this study was to evaluate the antiarrhythmic efficacy and mechanisms of action of dofetilide in an experimental model of atrial fibrillation. METHODS AND RESULTS: Dofetilide was administered intravenously to seven open-chest dogs with acute sustained atrial fibrillation induced by rapid atrial pacing for up to 4 hours. Mean atrial effective refractory period (ERP), dispersion of ERP, conduction velocity and wave-length were determined by multipoint right atrial programmed stimulation and activation mappin gusing a 56-electrode mapping plaque on the right atrial free wall. Dofetilide prolonged average ERP by 22% from 104 +/- 13 to 127 +/- 15 ms (P <.001), prolonged maximum ERP by 11% from 129 +/- 7 to 143 +/- 10 (P <.003), had no effect on conduction velocity at 200 ms pacing cycle length, slowed conduction velocity by 16% from 0.89 +/- 12 to 0.75 +/-.17 ms at 150 ms pacing cycle length, slowed conduction velocity by 16% from 0.89 +/- 12 to 0.75 +/-.17 ms at 150 ms pacing cycle length (P <.001), increased wavelength by 20% from 93 +/- 7 to 112 +/- 9 mm (P <.01), reduced dispersion of ERP by 24% from 11.4 +/- 2.9 to 8.7 +/- 2.3 (P =.016), and reduced the number of adjacent electrodes with ERP difference >/=20 ms by 67% from 18.4 +/- 7.1 to 6.1 +/- 4.2 (P <.001). Dofetilide reduced the number of excitation wavelets (total over three beats) entering the region of the mapping plaque by 38% from 5.0 +/-.8 to 3.1 +/-.4 (P <.002). Dofetilide terminated atrial fibrillation in all seven dogs at a mean of 3.4 +/- 2.2 minutes into the loading infusion and prevented reinduction of atrial fibrillation in all seven dogs after completion of the loading infusion, while on maintenance infusion. Time to termination of atrial fibrillation correlated closely with change in ERP (r = 0.78, P =.036). CONCLUSIONS: Dofetilide was highly effective in terminating and suppressing sustained pacing induced atrial fibrillation in this canine model. Time to termination of atrial fibrillation correlated with the degree of change in ERP produced by dofetilide. The mechanism of termination of atrial fibrillation by dofetilide appeared to be a progressive reduction and eventual extinction of re-entrant wavelets. The predominant electrophysiologic effects of dofetilide were prolongation of ERP and wavelength and a reduction in dispersion of refractoriness. Dofetilide had little effect on conduction velocity in this model, except at very short pacing cycle lengths. PMID- 10684459 TI - Modulation of the Electrophysiologic Actions of E-4031 and Dofetilide by Hyperkalemia and Acidosis in Rabbit Ventricular Myocytes. AB - BACKGROUND: E-4031 and dofetilide are new class III antiarrhythmic agents that inhibit the rapid component of the delayed rectifier potassium channel (I(Kr)); however, the effectiveness of many antiarrhythmic drugs in ischemic conditions is uncertain. METHODS AND RESULTS: We modeled two components of ischemia, hyperkalemia (9.6 mM) and acidosis (pH 6.8), in voltage-clamped single rabbit ventricular myocytes to help determine the effect of ischemia on the action of these two drugs. In physiologic solution both E-4031 and dofetilide blocked I(Kr) and significantly reduced total outward current. In hyperkalemic solution, both E 4031 and dofetilide showed significantly reduced blockade of I(Kr), while in acidotic solution dofetilide showed significantly reduced blockade of I(Kr) and E 4031 showed a trend to reduced blockade. Neither drug significantly reduced total outward current in hyperkalemic or acidotic solutions. CONCLUSIONS: In these conditions, E-4031 and dofetilide demonstrate reduced blockade of I(Kr), resulting in loss of class III effect. Furthermore, the complete loss of blocking effect on total outward current during simulated ischemia suggests increases of other repolarizing currents also contribute to loss of class III effect. PMID- 10684460 TI - Molecular Delivery System for Antisense Oligonucleotides: Enhanced Effectiveness of Antisense Oligonucleotides by HVJ-liposome Mediated Transfer. AB - BACKGROUND: The effectiveness of antisense oligodeoxynucleotides for in vitro and in vivo studies is limited by a low efficiency of cellular uptake and instability due to degradation by nucleases. To overcome some of these problems, we recently developed a transfer method that utilizes inactivated Sendai virus (hemagglutinating virus of Japan [HVJ]) complexed with liposomes to deliver antisense oligodeoxynucleotides. In this study, we compared the effectiveness of the HVJ-liposome method versus a cationic liposome method versus passive uptake to deliver antisense oligodeoxynucleotides against basic fibroblast growth factor on angiotensin (Ang) II-induced rat vascular smooth muscle cell growth. METHODS AND RESULTS: Twenty to twenty-eight hours after transfection, antisense fibroblast growth factor oligodeoxynucleotides introduced by passive uptake and HVJ-liposome method decreased basal DNA synthesis significantly as compared to the sense, control, and scrambled oligodeoxynucleotides groups; however, 60-68 hours after transfection, only antisense fibroblast growth factor oligodeoxynucleotides transduced by the HVJ-liposome method resulted in a significant inhibition of DNA synthesis under basal and Ang II-(10(-;6)M) stimulated conditions. The IC(25) of oligodeoxynucleotides assessed by the inhibition of thymidine incorporation was significantly lower using the HVJ liposome method than those using the other transfer methods. To clarify the mechanisms of cellular uptake of oligodeoxynucleotides with the HVJ-liposome method, we studied the cellular fate of fluorescein isothiocyanate (FITC)-labeled oligodeoxynucleotides FITC-oligodeoxynucleotides was localized in nuclei at 5 minutes after transfection with the HVJ-liposome method. In contrast, FITC oligodeoxynucleotides introduced by passive uptake was detected in nonnuclear cellular compartments, possibly endosomes, but not the nuclei. Cellular fluorescence of oligodeoxynucleotides introduced by passive uptake disappeared within 24 hours, while that introduced by the HVJ-liposome method could be observed up to 72 hours. CONCLUSION: These results demonstrate that the HVJ liposome transfer enhanced the effectiveness of AS-fibroblast growth factor via its specific molecular mechanisms of transfer. PMID- 10684461 TI - Mechanical Ablation of Concealed Left Lateral Bypass Tract. AB - A 50-yaer-old man with hypertension had been treated for supraventricular tachycardia with several medications for nine years. In 1990, he was started on amiodarone but a year later he developed side effects causing discontinuation of amiodarone. Because of his recurrent episodes of palpitations associated with near syncope, chest pain and shortness of breath, he underwent an electrophysiology study in 1992 that showed orthodromic AVRT with the presence of a concealed left-sided accessory bypass tract. Scheduled for radiofrequency ablation the following day, after catheters were placed and during mapping of the lateralmitral annulus, his tachycardia stopped abruptly without further inducability. Isoproterenol infusion during atrial and ventricular stimulation also failed to induce his original tachycardia. A year later, the patient presented with palpitations that felt different than his previous experiences. Work-up at that point only revealed a parasystolic focus on a 24-hour ECG monitoring without any form of supraventricular tachycardia. This represents a very unusual case by which the left lateral accessory pathway was mechanically ablated with catheter manipulation. This led to the disappearance of the orthodromic tachycardia that was easily induced before due to the activity of his parasytolic focus. The latter continued for the following four years but the patient has had no recurrences of his tachycardia. PMID- 10684463 TI - The Multicenter Automatic Defibrillator Implantation Trial. PMID- 10684462 TI - The Multicenter Automatic Defibrillator Implantation Trial. PMID- 10684464 TI - Searching for the Ideal Class III Antiarrhythmic Agent: How Pure Should they Be? PMID- 10684465 TI - Atorvastatin, a New HMG-CoA Reductase Inhibitor, Does Not Affect Glucocorticoid Hormones in Patients With Hypercholesterolemia. AB - BACKGROUND: Atorvastatin calcium (Lipitor) is a new 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor. The present study was conducted to examine the effect of pronounced cholesterol lowering on adrenal function in patients with severe hypercholesterolemia. METHODS AND RESULTS: Adrenal function was examined under basal conditions and following adrenal corticotropin hormone stimulation in 40 patients (36 with heterogeneous familial and 4 with polygenic hypercholesterolemia). The study was part of a larger study comparing the efficacy and safety of atorvastatin, colestipol, atorvastatin + colestipol, and simvastatin + colestipol treatment over a 1-year period. Maximum doses of all agents were studied: 80 mg atorvastatin once daily, 40 mg simvastatin once daily, and 20 g/day colestipol. At the end of the 1-year treatment period, reductions in low-density lipoprotein cholesterol were 57%, 54%, and 49% for the atorvastatin, colestipol + atorvastatin, and colestipol + simvastatin groups, respectively. No clinically significant changes in basal serum cortisol levels were seen in any treatment group during the 1-year treatment period. Mean serum cortisol concentrations and area under the curve for cortisol concentration versus time data following adrenal corticotropin hormone stimulation were not clinically different during treatment compared with values obtained at baseline for any of the treatment groups. CONCLUSIONS: Treatment with maximum doses of atorvastatin for 1-year did not have any adverse effects on adrenal function under basal conditions or during maximum stimulation. Similarly, colestipol therapy alone and in combination with either atorvastatin or simvastatin did not appear to affect adrenal function. PMID- 10684466 TI - Intravenous Propafenone for Conversion of Atrial Fibrillation or Flutter to Sinus Rhythm: A Randomized, Placebo-controlled, Crossover Study. AB - BACKGROUND: Propafenone has been claimed to be effective in converting atrial fibrillation and flutter to sinus rhythm; however, controlled clinical trials have reported variable results, and data about the safety of propafenone in the setting of heart failure are lacking. The aim of the present study was to evaluate the efficacy and safety of intravenous propafenone in converting atrial fibrillation and flutter to sinus rhythm. METHODS: Sixty patients with acute (<72 h) or chronic atrial fibrillation or flutter were included in a randomized, placebo-controlled, conditional cross-over study. Twenty eight patients, of whom 12 were in New York Heart Association class III and IV, had heart failure. Patients received intravenous propafenone (2 mg/kg in 10 minutes) and placebo subsequently at 1 hour intervals if sinus rhythm was not achieved. The patients' rhythms were continuously monitored for 1 hour and a 12-lead electrocardiogram, a 1-minute continuous rhythm strip and vital signs were recorded at baseline and at 15, 30, 45, and 60 minutes after the administration of each drug. RESULTS: Twenty of teh 59 patients (34%) treated with propafenone converted to sinus rhythm, while only 4 of the 50 patients (8%) treated with placebo converted (P <.001). Propafenone was more effective in patients with acute (<72 h) atrial fibrillation (64.5%). The conversion rate with propafenone was not significantly different from placebo in patients with atrial flutter and chronic atrial fibrillation (>72 h). Propafenone significantly decreased (P <.005 vs placebo) mean ventricular rate in nonresponders with a baseline heart rate of more than 100 beats/min. No clinically significant adverse effect occurred. CONCLUSIONS: We conclude that intravenous propafenone treatment is effective for converting acute atrial fibrillation; however, it seems unlikely to be beneficial in atrial flutter and chronic atrial fibrillation. Propafenone decreases ventricular rate in nonresponders, and a single dose of propafenone is relatively safe even in moderate-to-severe heart failure. PMID- 10684467 TI - Sotalol Is More Powerful Than Propranolol in Suppressing Complex Ventricular Arrhythmias. AB - BACKGROUND: Sotalol has combined type II and type III antiarrhythmic properties. Although the beta-blocking action of sotalol is thought to contribute to its antiarrhythmic actions, few data are available from direct comparative clinical trials with pure beta-blocking drugs. METHODS AND RESULTS: In this double-blind, randomized, multicenter, placebo-controlled, parallel study, we have compared the antiarrhythmic efficacy and safety of treatment with sotalol vs propranolol in 181 patients with organic heart disease and frequent (>30 ventricular premature complexes [VPCs]/h) repetitive ventricular premature complexes. Eighty-seven were randomized to receive sotalol and 94 received propranolol. The demographic and clinical characteristics of the two groups were identical, and the majority of patients had coronary artery disease or hypertensive heart disease. Most patients had a long-standing history (>5 years) of ventricular arrhythmias and, in a significant proportion, antiarrhythmic therapy with other drugs had failed in the past. After withdrawal of all antiarrhythmic drugs and 1 week of placebo, qualified patients were randomized to sotalol (320 mg/day) or propranolol (120 mg/day). patients not achieving adequate response were given higher doses of sotalol (640 mg/day) or propranolol (240 mg/day)At baseline, both groups had comparable frequency of total VPCs/hour (274/h and 255/h for sotalol and propranolol groups, respectively) which was reduced to 71 VPCs/h and 109/VPCs/h, respectively, at the end of phase 1. At final evaluation there was a significantly greater response to sotalol as demonstrated by 80% reduction in VPCs/hour with sotalol compared with only 50% reduction noted in the propranolol group. Adequate therapeutic response was also achieved in a significantly greater percentage of patients on sotalol compared with propranolol (56% vs 29%, P =.02). Sotalol was also superior to propranolol in suppressing the VT events/day during phase 1 (89% vs 78% reduction in VT events/day, P <.05). Sotalol was more effective than propranolol in all subgroups and in patients with heart rate <75 beats per minute. CONCLUSIONS: Sotalol is more powerful than propranolol in suppressing ventricular arrhythmias documented on Holter recordings. The superiority of sotalol appears to be related to its combined class II and class III antiarrhythmic actions. PMID- 10684468 TI - Frequency-dependent Cardiac Electrophysiologic Effects of Tedisamil: Comparison With Quinidine and Sotalol. AB - BACKGROUND: Tedisamil is a potent bradycardiac/antiischemic drug known to lengthen cadiac repolarization by blocking various potassium channels. Recent in vivo experiments revealed that it is an antiarrhythmic agent. It was therefore of interest to compare the cellular electrophysiologic effects of tedisamil with those of quinidine and sotalol in isolated cardiac preparations. METHODS AND RESULTS: The conventional microelectrode technique was applied in isolated dog cardiac Purkinje and ventricular muscle fibers and in rabbit left atrial muscle. Tedisamil (1 uM) and sotalol (30 uM) lengthened, while quinidine (10 uM) shortened action potential duration in dog Purkinje fibers. The phase 1 repolarization was delayed by tedisamil and quinidine and not changed by sotalol. In dog ventricular muscle and in rabbit atrial muscle, all three drugs studied lengthened repolarization. In dog Purkinje fiber, tedisamil and sotalol lengthened action potential duration more at slow than at high stimulation frequency (reverse use-dependence). In dog ventricular muscle fibers, the effect of the drugs was not clearly frequency dependent. In rabbit atrial muscle fibers, the quinidine-evoked repolarization lengthening was most pronounced at intermediate cycle lengths (500-1000 ms). Tedisamil and quinidine but not sotalol depressed the maximal rate of depolarization (V(max)), which depended on the stimulation frequency (use-dependence). The nature of the use-dependent V(max) block differed between quinidine and tedisamil. Quinidine decreased V(max) at a relatively wide range of stimulation frequencies whle tedisamil. Quinidine decreased V(max) at a relatively wide range of stimulation frequencies while tedisamil decreased V(max) largely at high rate of stimulation. Tedisamil and quiinidine prevented or decreased the pinacidil-evoked action potential shortening in dog ventricular muscle, suggesting block of the ATP-dependent potassium channels (I(KATP)), while with sotalol such effect was not observed. CONCLUSIONS: Although tedisamil, quinidine, and sotalol are known to lengthen the QT interval, their cellular electrophysiologic effects substantially differ. Tedisamil lengthens repolarization and prevents pinacidil-evoked action potential duration shortening, suggesting I(K(ATP)) blockade. Its effect on the V(max) is limited mostly to fast heart rate. These electrophysiologic effects of tedisamil resemble those of chronic amiodarone treatment. PMID- 10684469 TI - Nisoldipine Cardioplegia in the Isolated Rabbit Heart. AB - BACKGROUND: The metabolic and hemodynamic effects of nisoldipine supplementation in cardioplegia after ischemic injury were investigated in 13 isolated rabbit hearts. Group 1 consisted of 6 hearts, which received St. Thomas II cardioplegic solution. In group 2, nisoldipine was added to the cardioplegic solution at a concentration of 0.1 mg/kg in 7 hearts. METHODS: The explanted hearts were suspended from Langendorff apparatus and were perfused with Krebs-Henseleit solution. Left ventricular pressure, heart rate, malondialdehyde, glutathione peroxidase, glutathione reductase, reduced glutathione, oxidized glutathione, creatine kinase MB, (CK-MB), aspartate transaminase, and lactate dehydrogenase (LDH) were measured before and after 60 minutes of ischemia. Peak generated pressure after ischemia was significantly higher in group 2 versus group 1 while end-diastolic pressure was significantly lower in group 2 after ischemic arrest (P <.05). RESULTS: Malondialdehyde levels were lower in group 2 (P <.05). Glutathione peroxidase and glutathione reductase levels were significantly higher in group 2 (P <.05). The only enzymatic significant difference was observed between the preischemic and postischemic levels of aspartate transaminase in group 2 (P <.05). CONCLUSIONS: These findings show beneficial effects of nisoldipine cardioplegia, although its use as a cardioplegic additive is not yet possible. We believe, however, the effects of oral nisoldipine before cardiac surgery can be studied in a clinical setting. PMID- 10684470 TI - Cardiac Arrhythmias Following Intravenous Nicotine: Experimental Study in Dogs. AB - BACKGROUND: Nicotine, the active agent in tobacco, is released into the circulation during cigarette smoking. It elevates plasma catecholamines, heart rate, and arterial blood pressure; produces coronary spasm; and increases myocardial work and oxygen demand with concomitant reduction in oxygen supply. This may generate cardiac arrhythmias that might contribute to an increased incidence of sudden death due to smoking. It is hypothesized that acute administration of nicotine will induce cardiac arrhythmias, and this experimental study was planned with an aim to assess arrhythmogenic activity as a result of acute administration of nicotine. METHODS: Nicotine was administered in different doses intravenously in 16 anesthesized dogs, and 52 experiments were carried out at weekly intervals. In each experiment, continuing anesthesia and after nicotine administration. They were scrutinized by two experienced electrocardiographers at intervals of 1, 2, 3, 4, 5, 10, 15, and 30 minutes. RESULTS: Data revealed nonsignificant arrhythmias with doses of 2.5, 5.0, and 10.0 mg/kg of intravenous nicotine. The dose of 50 ug/kg induced supraventricular arrhythmias, atrioventricular junctional arrhythmias, and ventricular arrhythmias. Supraventricular bradycardia in 30 (83%; P <.0001), supraventricular arrhythmia in 30 (83%; P <.0001), sinus arrest in 18 (50%; P <.003), atrial ectopics in 24 (67%; P <.0004), and atrial tachycardia in 98 experiments (25%; P <.021). These results were statistically significant. In 18 experiments, sinus arrest was observed to be missing P waves and QRS complexes for a period corresponding to 4:1-10:1 SA block, lasting 2-6 seconds, within 3 seconds of injection. Occurrence of wandering pacemaker was observed in 6 experiments, atrial flutter in 2, and atrial fibrillation in 2, but these incidents were not significant. Atrioventricular junctional arrhythmias consisted of escape beats in 9 subjects (25%; P <.02), premature contractions in 12 (33%; P <.005), first-degree heart block in 9 (25%; P <.02), second degree heart block in 9 (25%; P <.02) and atrioventricular dissociation in 9 (25%; P <.02). All arrhythmias in this category were significant. Ventricular arrhythmias consisted of ventricular premature contractions that were unifocal in 32 subjects (89%; P <.0001), multifocal in 30 (83%; P <.0001), bigeminy in 28 salvos in 18 (50%; P <.003). Sustained ventricular tachycardia (> 30 beats) in 12 experiments (33%; P <.005) proved significant. The dose of 100 ug/kg induced fatal ventricular flutter and ventricular fibrillation. The dog expired and experiments with that dose were not repeated. CONCLUSION: Data reveal dose-dependent arrhythmogenecity of nicotine in dogs. Smaller doses of nicotine did not produce significant arrhythmias. Higher doses, bioequivalent to smoking two standard cigarettes, may generate cardiac arrhythmias of simple to severe nature. Further work in human beings may confirm whether nicotine in cigarette smoke will generate similar cardiac arrhythmias especially in patients with autonomic imbalance and/or compromised and ischemic myocardium. PMID- 10684471 TI - Adrenergic-dependent Effect of Adenosine-induced Ventricular Fibrillation in the Isolated Rabbit Heart. AB - BACKGROUND: The present study examined the contributory role of endogenous catecholamines in adenosine-induced ventricular fibrillation in isolation rabbit hearts. METHODS AND RESULTS: Cardiac catecholamine depletion was induced in eleven rabbits by the administration of 6-hydroxydopamine (2 x 30 mg/kg, every 12 hours intramuscularly). Hearts were removed 24 hours later, and subjected to 12 minutes of hypoxic perfusion followed by 40 minutes of reoxygenation while heart rate was maintained with atrial pacing. One of six, and one of five hearts from 6 hydroxydopamine treated rabbits developed ventricular fibrillation during hypoxia reoxygenation when exposed to 3,7-dimethyl-1-propargylzanthine (DMPX) (10 uM) + adenosine (ADO) (1 uM) and DMPX (10 uM) + ADO (10 uM), respectively. In hearts from a control group, not exposed to 6-hydroxydopamine, ventricular fibrillation developed in each of five (100% incidence) hearts when perfused in the presence of DMPX (10 uM) + ADO (10 uM) (P <.05). Nadolol (1 uM), a beta-adrenoceptor DMPX (10 uM) + ADO (10 uM) treated hearts (n = 6, P <.05 vs DMPX + ADO treated hearts). To ensure catecholamine depletion, spontaneously beating isolated hearts from vehicle and 6-hydroxydopamine treated rabbits were perfused under normoxic conditions while exposed to increasing concentrations of tyramine (1, 3, 10 mM) and the change in heart rate was determined. A concentration-related, positive chronotorpic response to tyramine was obtained in hearts from the vehicle treated group that was absent in hearts from 6-hydroxy-dopamine treated rabbits or hearts perfused in the presence of nadolol. CONCLUSIONS: The results demonstrate that inhibition of the cardiac adenosine A(2) receptor, unmasks an adenosine A(1) receptor profibrillatory effect that is dependent upon endogenous cardiac catecholamines and beta-adrenoreceptor activation during myocardial hypoxia reoxygenation. PMID- 10684472 TI - Prevention of Hypercholesterolemic Atherosclerosis by Garlic, an Antixoidant. AB - BACKGROUND: Investigations of the effects of high cholesterol diet in the presence and absence of garlic on the genesis of atherosclerosis, the blood lipid profile, aortic tissue lipid peroxidation product malondialdehyde, chemiluminescence, a marker for antioxidant reserve and activity of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase were made in rabbits. METHODS AND RESULTS: Four groups of 10 rabbits each were studied: group 1 was given regular rabbit chow, group 2 was given rabbit chow diet supplemented with garlic powder (300 mg twice daily orally), group 3 was given 1% cholesterol diet, group 4 was given 1% cholesterol diet supplemented with garlic powder (300 mg twice daily orally). Blood concentration of triglyceride, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and very low-density lipoprotein cholesterol were measured before and after 4 and 10 weeks of experimental diets. The aorta was removed at the end of protocol (10 weeks) for assessment of atherosclerotic changes (gross and microscopic), malondialdehyde concentration, chemiluminescence, and activity of antioxidant enzymes. Total cholesterol, low density-lipoprotein cholesterol and ratio of low density lipoprotein cholesterol/high-density lipoprotein cholesterol and ratio of low-density lipoprotein cholesterol/high-density lipoprotein cholesterol increaserd in group 3 and 4; the increase was smaller in group 4 than in in group 3 although not significant. Serum high-density lipoprotein cholesterol decreased to a similar extent in groups 3 and 4. Serum triglyceride and very low-density lipoprotein cholesterol remained unchanged in group 3 but increased in group 4. These values were significantly higher than those in group 1. Garlic in rabbits with control diet decreased the levels of triglyceride and very low density lipoprotein but did not affect the levels of total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol and the ratio of low-density lipoprotein cholesterol/high-density lipoprotein cholesterol. There was an increase in aortic tissue malondialdehyde, chemiluminescence, and activities of catalase and glutathione peroxidase in group 3 compared with those in group 1. Levels of aortic malondialdehyde, chemiluminescence, catalase, and glutathione peroxidase were lower in group 4 compared with group 3; however, values for malondialdehyde and chemiluminescence were lower and that of catalase and glutathione peroxidase were higher in group 4 compared with group 1. Superoxide dismutase activity was similar in all the four groups. Malondialdehyde, chemiluminescence, and activity of catalase of aortic tissue decreased while activity of glutathione peroxidase increased in group 2. Atherosclerotic changes were lower in group 4 compared with group 3. Histologic changes were practically similar in groups 3 and 4. CONCLUSIONS: Increased levels of malondialdehyde, chemiluminescence, and antioxidant enzymes associated with development of atherosclerosis suggests a role for oxygen free radicals in the pathogenesis of hypercholesterolemic atherosclerosis. The protection afforded by garlic was associated with decrease in aortic malondialdehyde and chemiluminescence inspite of no change in serum cholesterol. These findings suggest that oxygen free radicals are involved in the genesis and maintenance of hypercholesterolemic atherosclerosis and that use of garlic can be useful in preventing the development of hypercholesterolemic atherosclerosis. PMID- 10684473 TI - Mibefradil: A New Selective T-Channel Calcium Antagonist for Hypertension and Angina Pectoris. AB - Calcium antagonists are an established therapy for patients with hypertension and angina pectoris, but their current usage is often limited by their pharmacologic profilers and side effects. Mibefradil is a recently developed calcium antagonist with a unique chemical structure, site of action, and set of pharmacologic effects. Unlike currently available calcium channels as well as L-type channels. It is further distinguished from the other calcium antagonists in that it is the first member of a new class of calcium antagonists, the tetralol derivatives. With chronic oral dosing, mibefradil attains steady-state plasma concentrations within 3-4 days, has a bioavailability of approximately 90%, and a plasma half life of 17-25 hours. It has a gradual onset of action and can be administered once daily without regard to food intake. It increases coronary blood flow and lowers peripheral vascular resistance. The vasodilatory effects of mibefradil are associated with a lack of inotropic effect on myocardium, lack of neurohormonal activation, and a reduction in heart rate. In clinical trials it has been demonstrated to be an effective agent in the treatment of patients with hypertension and angina pectoris, with a good safety and tolerability profile regardless of age, gender, or race. PMID- 10684474 TI - A Therapeutic Commentary. AB - ABSTRACT: Thirty-one million patients in the United States undergo surgical procedures every year. Approximately 10%-the majority of these with hypertension are at an increased risk for perioperative and postoperative cardiovascular morbidity and mortality. Thus, hypertensive patients requiring surgery, especially the 2.1 million undergoing noncardiac procedures, should be evaluated carefully for the magnitude, and if severe, the cause of the hypertension. Additionally, their associated metabolic and cardiovascular status should be characterized and corrected with aggressive therapy. Hypertensive patients with known ischemic heart disease, those with multiple risk factors for ischemic heart disease (IHD), some with valvular heart disease, and those with congestive heart failure should be evaluated for their ability to perform the physical and social activities of everyday life, and, when necessary, have formal stress testing. Most studies suggest that blood pressures of 180/110 mm Hg or greater are associated with a greater risk for perioperative ischemic events. Therefore, the goals of blood pressure control should be to reduce the blood pressure without jeopardizing organ function. Antihypertensive medication should be administered until the time of surgery. beta-Receptor blockers should be instituted or continued in patients with angina and in some patients with congestive heart failure. Those without prior antihypertensive therapy might be best treated with beta-blocker therapy perioperatively as evidenced by the Multicenter Study of Perioperative Research Group with atenolol and those earlier studies with metoprolol. The risks of the surgery should be discussed with the patient so the risks can be weighed against the expected benefit. Studies suggest that perioperative risk for any patient, and especially patients with hypertension, are in part related to the adrenergic arousal before, during, and after the procedure as evidenced by the rise in heart rate and blood pressure, along with the liberation of clotting facators and increased risk for plaque rupture, coronary vasoplasm, and consequent myocardial infarction and fibrosis. PMID- 10684475 TI - Angiotensin II Receptor Regulates Ionic Currents in Guinea Pig Ventricular Myocytes. AB - BACKGROUND: Previous studies have shown that angiotensin II (Ang II) receptors are preset in a wide variety of target tissues and that Ang II regulates the target tissue functions through Ang II receptors. However, the action of Ang II receptors on transsarcolemmal currents in ventricular myocytes has not been elucidated. METHODS AND RESULTS: We performed whole-cell voltage clamp and patch clamp experiments to determine the effects of Ang II-receptor agonists and antagonists on ionic currents in single isolated guinea pig ventricular myocytes. We found that extracellular perfusion of Ang II (30 nM) increased the L-type Ca(2+) current from 581 +/- 27 to 837 +/- 42 pA (n = 5, P <.01). Ang II also prolonged the Ca(2+) current activation and inactivation time constants. These were reversible by losartan (100 nM), a type 1 Ang II receptor (AT(1)) blockade. On the other hand, perfusion of 30 nM Ang II decreased K(+) current (I(K)) from 1543 +/- 28 to 1194 +/- 50 pA (n = 5, P <.05) and K(+) tail current (I(K-tail)) from 275 +/- 24 to 206 +/- 29 pA (n = 5, P <.05). These effects were also abolished by perfusion of losartan. However, perfusion of Ang II resulted in an increase of inward rectified K(+) current (I(K1)) in whole-cell recordings. Single channel recordings showed that the increase in I(K1) was attributed to a burst opening current with a larger unit of amplitude. These effects were reversed by saralasin but not losartan, indicating possible type 2 Ang II receptor (AT(2)) involvement. CONCLUSIONS: Our results provide evidence that Ang II receptors regulate the transsarcolemmal currents in single guinea pig ventricular myocytes. Therefore, Ang II regulation of ionic currents is mediated through the different subtypes of Ang II receptors. PMID- 10684476 TI - Electrocardiographic Changes and Mortality Due to Myocardial Infarction in Rats With or Without Imidapril Treatment. AB - BACKGROUND: Various angiotensin-converting enzyme inhibitors are known to improve heart function and prolong survival in patients and animals after myocardial infarction. Because myocardial infarction is known to induce arrhythmias, this study tested the hypothesis that early treatment with the angiotensin converting enzyme inhibitor imidapril reduces mortality during acute myocardial infarction because of protective effects against arrhythmogenesis. METHODS AND RESULTS: Rats were randomly divided into four groups: sham control, myocardial infarction, sham plus imidapril, and myocardial infarction plus imidapril. Myocardial infarction was produced by ligation of the left anterior descending coronary artery. Treated rats received imidapril (1 mg/kg/day) through a gastric tube beginning 1 hour after coronary occlusion; control rats received tap water. Electrocardiogram (ECGs) were recorded 1, 3, 7, and 21 days postocclusion. Infarct size and scar weight were determined at 21 days in the myocardial infarction groups with and without imidapril treatment. ECGs of untreated rats showed ST-segment changes, abnormal Q waves, premature ventricular complexes, and QT(c) prolongation 1-21 days after coronary occlusion. Total mortality in 21 days averaged 35% in untreated rats; mortality within 48 hours was 30%. On the other hand, imidapril treated rats showed fewer ST-segment changes, fewer abnormal Q waves, and a decreased incidence of premature ventricular complexes after coronary occlusion; the ST-segment and QT(c) interval returned to basal values within 1 week after occlusion. Imidapril treatment did not affect the ECG pattern in sham-treated control animals. Total mortality in the imidapril-treated group in 21 days after infarction was 22.5%; mortality within 48 hours was 20% (P <.05 compared with the untreated infarction group). Infarct size and scar weight caused by coronary occlusion did not differ in the untreated and imidapril-treated groups. CONCLUSIONS: Early treatment with imidapril markedly decreases mortality in rats after acute myocardial infarction. The lower mortality is not associated with a decrease in infarct size but is consistent with a protective effect of the drug against arrhythmogenesis. PMID- 10684477 TI - Platelet-Mediated Cardioprotective Effect Against Ischemia-Reperfusion Injury in Isolated Rat Hearts: Role of Platelet Number and Contribution of Supernatant of Aggregated Platelets. AB - BACKGROUND: Previous studies have documented cardioprotective effects of circulating platelets after reperfusion injury. The present study was designed to examine the role of platelet number and contribution of platelet-released mediators in the platelet supernatant in cardioprotection against ischemia reperfusion-induced myocardial dysfunction. METHODS AND RESULTS: Isolated buffer perfused (constant volume) Sprage-Dawley rat hearts were subjected to 25 minutes of global ischemia followed by 30 minutes of reperfusion. Ischemia-reperfusion resulted in myocardial dysfunction, indicated by an increase in coronary perfusion pressure and left ventricular end-diastolic pressure, and a decrease in developed left ventricular pressure. Perfusion of hearts with washed rat platelets (10(3)-2.2 x 10(7) cells/mL) significantly (P <.01) attenuated these indices of myocardial dysfunction upon ischemia-reperfusion in a concentration dependent manner. A cardioprotective effect of platelets was observed at a concentration as low as 10(5) platelets/mL. Similar cardioprotection was seen in hearts perfused with the supernatant of aggregated platelets. CONCLUSIONS: These observations indicate that the platelet-mediated cardioprotective effect against ischemia-reperfusion in vitro is concentration dependent, and platelet-released mediators in the platelet supernatant are protective against ischemia-reperfusion injury. PMID- 10684478 TI - Comparative Study of the Effects of Erythromycin and Roxithromycin on Action Potential Duration and Potassium Currents in Canine Purkinje Fibers and Rabbit Myocardium. AB - BACKGROUND: Erythromycin and roxithromycin are macrolide antibiotics in common clinical use. Erythromycin occasionally produces life-threatening arrhythmias (torsades de pointes) by blocking the outward potassium current responsible for repolarization of the cardiac action potential. METHODS AND RESULTS: We used standard cellular electrophysiological and whole-cell patch-clamping techniques to compare the relative efficacy of erythromycin and roxithromycin in prolonging cardiac action potential in canine Purkinje fibers and in blocking individual outward potassium currents in isolated rabbit ventricular myocytes. We demonstrated significant prolongation of action potential duration in canine Purkinje fibers by erythromycin but not roxithromycin at a concentration of 100 uM. The delayed rectifier, the outward potassium current thought to be most sensitive to modulation by drugs, was significantly depressed by both agents at concentrations of >/=30 uM in isolated rabbit ventricular myocytes. Both drugs had similar potencies (26% and 21% reduction by 30 uM erythromycin and roxithromycin, respectively, and 50% and 36% reduction by 100 uM erythromycin and roxithromycin). Neither agent significantly blocked other potassium currents (including the transient outward current). CONCLUSIONS: Taking into account normally observed peak blood concentrations of these agents in clinical use and the fact that roxithromycin is not normally administered intravenously, we conclude that the risk of proarrhythmia during normal clinical use of oral roxithromycin is extremely remote. PMID- 10684479 TI - Lysophosphatidylcholine and Cellular Potassium Loss in Isolated Rabbit Ventricle. AB - BACKGROUND: Lysophospholipids such as lysophosphatidylcholine (LPC) have many direct electrophysiological effects on cardiac muscle and have been implicated as a cause of lethal ventricular arrhythmias during acute myocardial ischemia. Because extracellular K(+) accumulation is also a key arrhythmogenic factor during acute ischemia, we examined the effects of LPC on cellular K(+) balance, including its interaction with adenosine triphosphate-sensitive K(+) (K(ATP)) channels. METHODS AND RESULTS: Isolated rabbit interventricular septa paced at 75 beats/min were loaded with (42)K(+) to measure unidirectional K(+) efflux rate (in (42)K(+) washout experiments) or tissue K(+) content ((42)K(+) uptake experiments) and action potential duration (APD) during exposure to 20 uM LPC for 30 minutes. LPC caused tissue K(+) content to decrease by 15 +/- 2% (n = 4) at a steady rate over 30 minutes, associated with gradual APD shortening and a delayed increase in unidirectional K(+) efflux rate. Pretreatment with 12 uM cromakalim to selectively activate K(ATP) channels shortened APD by 44 +/- 66% and had no effect on net tissue K(+) content during control aerobic perfusion. However, cromakalim increased net K(+) loss during exposure to LPC to 22 +/- 4%, a 47% increase. CONCLUSIONS: LPC induced net K(+) loss in heart, which was potentiated by the K(ATP) channel agonist cromakalim. This ATP finding suggests that if LPC accumulates to similar levels during myocardial ischemia and hypoxia, it may be an important mechanism in net K(+) loss. PMID- 10684480 TI - Prolonged Captopril Therapy in Murine Viral Myocarditis. AB - BACKGROUND: Acute myocarditis can progress to chronic heart muscle disease and cardiomyopathy. In the coxsackievirus B(3) (CB(3)) mouse model of myocarditis, early administration of captopril, an angiotensin-converting enzyme (ACE) inhibitor, ameliorated histopathological changes in inflammation, necrosis, and calcification and reduced heart weight. Late administration of captopril reduced heart weight but did not affect the histological findings. In this study, we investigated the effects of prolonged captopril treatment in the chronic phase of this model. METHODS AND RESULTS: Three-week-old male CD(1) mice were infected with CB(3) and then randomized to receive placebo or captopril starting on day 7 of infection. Captopril, 2 g/L, was given as the drinking water daily for up to 6 months. Autopsies were performed at 6 and 10 months. Heart-to-body weight ratios were obtained, and deaths were tallied. Myocardial fibrosis was graded according to a score system. In addition, picrosirius red stain (PSR) also was used for assessment of collagen deposition. Mean heart weights were similar in both groups. Mean body weight was significantly lower in captopril-treated mice (40.7 g) than in the untreated group (43.6 g) at 6 months (P =.0155), and mortality was higher (8.7 vs 0.87%; P =.009). At 6 months, the mean myocardial fibrosis score in treated mice (0.12) was significantly less than in untreated animals (0.35; P =.035). With PSR, the mean myocardial fibrosis score in the captopril group (1.20) was also significantly less than in the untreated group (1.58; P =.045). At 10 months, fibrosis scores were similar in both groups. CONCLUSIONS: Chronic captopril treatment in CB(3) myocarditis reduces myocardial fibrosis. PMID- 10684481 TI - Formation of Reactive Oxygen Species in Various Vascular Cells During Glyceryltrinitrate Metabolism. AB - BACKGROUND: Anti-ischemic therapy with organic nitrates as nitric oxide (NO) donors is complicated by the induction of tolerance. When nitrates are metabolized to release NO, there is a considerable coproduction of reactive oxygen species (superoxide radical and peroxynitrite) in vessels leading to inactivation of NO, to diminished cyclic quanosine monophosphate production in smooth muscle cells (SMC), to impaired vasomotor responses to the endothelium derived relaxation factor (EDRF), and to formation of nitrotyrosine as a marker of glyceryltrinitrate (GTN)-induced formation of peroxynitrite. The aim of the study was to analyze in vitro the formation of superoxide radicals and of peroxynitrite in GTN-treated endothelial and smooth muscle cells and in washed ex vivo platelets using electron spin resonance and spin-trapping techniques. METHODS AND RESULTS: Using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap, it was shown that in platelets, smooth muscle, and endothelial cells incubated acutely for 15 minutes with 0.5 mM GTN, the rate of generation of reactive oxygen species (ROS) was twice as high as under control conditions. Using the new spin-trap 2H-imidazole-1-oxide (TMIO), a GTN-induced peroxynitrite formation was detected in SMC and in platelets incubated with 0.5 mM GTN for 15 minutes. Spin-trap 1-hydroxy-3-carboxy-pyrrolidine (CP-H) was used to estimate the rate of ROS formation in platelets incubated for 15 minutes with 0.5 mM GTN; the rate amounted to 14.6 +/- 1.1 nM/min/mg protein compared with 4.0 +/- 0.4 nM/min/mg protein in controls. The rate of ROS formation in SMCs was substantially increased (240 +/- 16%) after initiation of GTN tolerance by treatment of the cells in culture with 100 uM GTN for 24 hours. CONCLUSIONS: GTN increases the formation of superoxide radicals in endothelial cells, SMCs, and platelets. Peroxynitrite is formed during GTN metabolism in vascular cells and may contribute to the development of tolerance. A decrease in the nitrate-induced inhibition of platelet aggregation during GTN tolerance is associated with oxidative actions of ROS formed in platelets during GTN metabolism. PMID- 10684482 TI - Infarct Size Reduction by Ischemic Preconditioning Is a Monophasic, Short-Lived Phenomenon in Anesthetized Pigs. AB - BACKGROUND: Controversy exists concerning the duration of infarct size reduction with ischemic preconditioning in different species. In the present study, we (a) evaluated the time course of protection with preconditioning and (b) sought to determine whether late protection (the "second window") after 24 hours is manifest in the open-chest pig model. METHODS AND RESULTS: Six groups of pentobarbital-anesthetized pigs underwent 1 hour of left anterior descending coronary artery occlusion and 2 hours of reperfusion. Group 1 served as control, and pigs in group 2 received two 10-minute episodes of preconditioning ischemia followed by 30 minutes of reperfusion before the sustained 1-hour occlusion. In groups 3-6, the period of intervening reperfusion between the preconditioning stimulus and the index ischemia was extended to 60, 90, and 300 minutes and 24 hours, respectively. The area at risk was determined by fluorescein dye injection, and infarct size was measured by incubation in p-nitrobluetetrazolium and expressed as percent of the risk area. Infarct size in preconditioned pigs (group 2) was significantly reduced compared with controls (25.6 +/- 3.9% v 71.3 +/- 5.9%, P <.001). Extension of the intervening reperfusion to 60, 90, and 300 minutes and 24 hours resulted in infarct sizes of 64.5 +/- 5.5%, 67.2 +/- 8%, 62.6 +/- 6.1%, and 75.3 +/- 7%, respectively (P = NS v control). CONCLUSIONS: The infarct size-limiting effects of ischemic preconditioning last less than 1 hour in the pig model. Moreover, in contrast to other species, a late protection at 24 hours after the preconditioning stimulus was not detected. These results indicate that precondition-induced reduction of infarct size is monophasic in anesthetized pigs. PMID- 10684483 TI - The Effect of Atorvastatin on the Human Lens After 52 Weeks of Treatment. AB - BACKGROUND: The effect of atorvastatin calcium (Lipitor, Parke-Davis, Morris Plains, NJ) on the crystalline lenses of hypercholesterolemic patients was evaluated and compared with that of lovastatin after 52 weeks of treatment to reduce cholesterol levels. METHODS AND RESULTS: Six hundred ninety-six atorvastatin-treated and 235 lovastatin-treated patients completed a large safety study that included an ophthalmologic examination. Efficacy was evaluated as mean percent change from baseline in low-density lipoprotein (LDL) cholesterol. Patients received atorvastatin, 10 or 20 mg, or lovastatin, 20 or 40 mg, once daily for either 36 or 52 weeks. Patients were evaluated by slit-lamp examination, and a standardized format was used to describe the findings. Best corrected visual acuity was measured using the Snellen chart. Patients treated with atorvastatin had significantly (P .01). CONCLUSION: Insulin resistance impairs endothelium-dependent relaxation in small mesenteric arteries. It appears that insulin resistance transforms the primary relaxant factor from endothelial-derived hyperpolarizing factor to nitric oxide. These findings suggest that hypertension and atherosclerosis associated with the IR syndrome are caused, at least in part, by endothelial dysfunction. PMID- 10684491 TI - Nitric Oxide Alters Human Microvascular Endothelial Cell Response to Cyclic Strain. AB - BACKGROUND: Nitric oxide (NO), a potent vasodilator and inhibitor of platelet function, is elaborated constitutively by endothelial cells through the oxidation of l-arginine by endothelial NO synthase. Although several biochemical agonists, such as bradykinin, have been shown to stimulate NO production by the endothelium, the effects of physical factors have been less well characterized. We have previously examined the shear-stress-dependent induction of NO production by the endothelium, and others have examined the effects of transmural pressure on NO production. In the current study, we analyzed the effects of cyclic strain or load in the presence and absence of an NO agonist on endothelial cell proliferation. METHODS AND RESULTS: Subconfluent human microvascular endothelial cells were grown on deformable culture plates in media containing 10% fetal bovine serum and 1 mM l-arginine, and then subjects to either 0, 11, 18, or 27% cyclic strain at a frequency of 1 Hz. Nitrogen oxides (S-nitrosothiols and free NO) in media were measured by photolysis-chemiluminescence at the end of 24 hours, as were cell number, [(3)H]thymidine incorporation, and total cell protein. In the absence of an NO agonist, 11% cyclic strain produced a 73 +/- 17% increase in nitrogen oxides compared with control (P <.05), whereas 18 and 27 cyclic strain produced no significant increase in nitrogen oxides compared with control. However, cells subjected to 27% strain increased cell number (by 98 +/- 17%) and [(3)H]thymidine incorporation (by 16 +/- 4%, P <.05) compared with control. In an attempt to examine further the effects of endogenous NO on the cells' proliferation response to increasing levels of cyclic strain, we incubated human microvascular endothelial cells with 1 uM bradykinin and subjected them to 27% cyclic strain. In the presence of bradykinin and 27% cyclic strain, production of nitrogen oxides and endothelial NO synthase activity were increased by 110 +/- 37% and 135 +/- 7%, respectively (P <.05 compared with control); however, we observed no significant increase in cell number (5 +/- 5%), despite an 86 +/- 7% increase in [(3)H]thymidine incorporation (P <.05). CONCLUSION: These results suggest that endogenous NO production by human microvascular endothelial cells regulates their growth response to cyclic strain. Modulation of NO production in vivo in areas of the vasculature that are subjected to increased levels of strain may represent one potential mechanism by which to alter endothelial cell proliferation. PMID- 10684492 TI - Protective Effects of Iganidipine on Morphological and Functional Changes of Arteries in Hypertensive Dahl Rats. AB - BACKGROUND: This study was performed to examine the protective effects of iganidipine, a new water-soluble calcium antagonist, on the morphological and functional changes of arteries in Dahl salt-sensitive (Dahl-S) rats. METHODS AND RESULTS: Vehicle and iganidipine were administered orally to Dahl-S rats fed a high-salt diet (HSD) for 8 weeks. Aorta, superior mesenteric arteries (SMA), and peripheral mesenteric arteries (PMA) were examined light-microscopically or electon-microscopically. Relaxant responses of isolated aorta and SMA were recorded isometrically. In rats fed HSD, blood pressure was markedly increased. Light microscopy showed intimal and medial hypertrophy, periarteritis, and narrowed arterial lumen in the PMA. Transmission and scanning electron microscopy or light microscopy showed medical thickness in the aorta and SMA and hypertrophy of endothelial cells and dilatation of the subendothelial space only in the aorta. In the SMA, both endothelium-dependent relaxation (EDR) and endothelium independent relaxations (EIR) were reduced to a similar extent. In the aorta, the EDR was more markedly attenuated than the EIR. Iganidipine at 3 mg/kg/day showed a 24-h sustained hypotensive effect and completely prevented the morphological and functional changes in both arteries. Iganidipine at 1 mg/kg/day, which lowered blood pressure only for several hours, decreased the injuries in PMA and aortic endothelium and moderately restored the EDR in the aorta. Iganidipine at 0.3 mg/kg/day had no effects. CONCLUSIONS: In Dahl-S rats fed an HSD, iganidipine completely prevented all the changes at a sustained-hypotensive dose and prevented the injuries of PMA and aortic endothelium and the reduction of EDR in the aorta at a nonsustained hypotensive dose. Nonhemodynamic effects of iganidipine may be partly involved in its protective effects against arterial injuries. PMID- 10684493 TI - The Effect of Melatonin on Hemodynamics, Blood Flow, and Myocardial Infarct Size in a Rabbit Model of Ischemia-Reperfusion. AB - BACKGROUND: Melatonin, a hormone, has gained popularity and is being used by millions for a variety of indications. There are few data on its safety or its effects on hemodynamics and coronary blood flow. Also, studies have confirmed that melatonin is a potent antioxidant. Therefore, it may be capable of scavenging free radicals during the reperfusion phase after a heart attack. This study evaluates the safety of melatonin with regard to its cardiovascular effects and tests the hypothesis that melatonin might be protective in the setting of ischemia-reperfusion and reduce myocardial infarct size. METHODS AND RESULTS: Anesthetized rabbits were treated with melatonin (n = 8, 10 mg/kg, intravenously) 10 minutes before coronary artery occlusion (CAO) and again 15 minutes before reperfusion. Control rabbits received vehicle (n = 8). All rabbits underwent 30 minutes of occlusion and 3 hour reperfusion. Both before and during CAO, melatonin did not alter heart rate compared with control (185 +/- 7 beats/min v 181 +/- 7 before; and 179 +/- 5 v 181 +/- 9 during, respectively, P = NS) or blood pressure (70 +/- 4 mmHg v 66 +/- 6 before and 59 +/- 4 v 58 +/- 5 during, respectively, P = NS). Regional myocardial blood flow (RMBF) was similar before CAO in the melatonin group (1.18 +/- 0.17 mL/min/g) versus the control group (1.15 +/- 0.10 mL/min/g). Infarct size, expressed as a fraction of ischemic risk zone, was similar in the melatonin (0.29 +/- 0.03) and control groups (0.29 +/- 0.06, P = NS). At a higher dose of 50 mg/kg in treated (n = 7) versus control (n = 7) rabbits, melatonin treatment did not alter heart rate (204 +/- 14 in melatonin group v 181 +/- 5 in controls, P = NS) or blood pressure (80 +/- 11 in melatonin v 66 +/- 7 in controls, P = NS) when compared with control. Melatonin at this dose also did not affect infarct size, 0.38 +/- 0.06, when compared with control, 0.34 +/- 0.07, P = NS. CONCLUSION: Melatonin's effects on hemodynamics and coronary blood flow were neutral, and it did not exacerbate myocardial ischemia or necrosis in this model. Melatonin appears to be a safe drug with no apparent effects on the cardiovascular system in this model. PMID- 10684494 TI - ACE Inhibitors and Renal Vascular Responses in the Spontaneously Hypertensive Rat. AB - BACKGROUND: Substantial evidence has accumulated for the intrarenal generation of functionally important quantities of angiotensin II (Ang II). To assess the possibility that Ang II generation occurs beyond a barrier to diffusion from the vascular compartment, six angiotensin-converting enzyme (ACE) inhibitors varying widely in their lipid solubility were employed in the spontaneously hypertensive rat (SHR) and their normotensive controls (WKY). The biological end points were renal blood flow and its response to Ang II. RESULTS: Two ACE inhibitors, ramipril and captopril, induced a larger increase in renal blood flow and enhanced the renal vascular response to Ang II substantially more than did enalapril and lisinopril. The two prodrugs, enalapril and ramipril, which are substantially more lipophilic than the respective active drugs, enalaprilat and ramiprilat, showed equivalent responses. The partial agonist saralasin virtually abolished the renal vasodilator response to ramipril. The pattern of response was similar in WKY, but the responses were substantially smaller. CONCLUSIONS: The results support the concept that a functionally important compartment for intrarenal Ang II formation exists in the healthy rat and that this process is amplified in the SHR. PMID- 10684495 TI - Clonidine-Induced Heat-Shock Protein Expression in Rat Aorta. AB - BACKGROUND: Restraint-stress and administration of drugs that precipitate hypertension induce heat-shock protein (HSP) expression in the aorta. The exact mechanism supporting this hypertension-related HSP response is unclear because HSP induction is blocked by receptor-selective and nonselective antihypertensive agents. METHODS AND RESULTS: To identify mechanisms contributing to the pharmacological/physiological regulation of the HSP response in cardiovascular tissues, we administered clonidine to awake and freely moving animals to determine its effect on HSP expression in vivo. Inconsistent with previous work, we found that clonidine produced a dose-dependent and transient increase in HSP70 mRNA levels in the aorta. No other tissue examined displayed an HSP response after clonidine administration. Clonidine-induced HSP expression was not restricted to the HSP70 family; HSP89alpha, HSP89beta, and HSP60 were also induced. Interestingly, no heat-shock element-binding activity was observed after clonidine administration, suggesting that unusual transcriptional regulatory mechanisms mediate this response. Yohimbine and nifedipine blocked HSP70 mRNA expression, whereas isoproterenol, mecamylamine, and reserpine had no effect. CONCLUSIONS: The functional consequence of HSP expression in cardiovascular tissues may be to alter the responsiveness of cells in these tissues to subsequent drug or stress exposures, thereby implicating the HSP response as an important component of cardiovascular homeostasis. If so, treatment of mammalian organisms with drugs capable of inducting selective HSP expression in vascular tissue may alter the progression of cardiovascular disease processes. PMID- 10684496 TI - Update on Atrial Fibrillation: Restoration of Sinus Rhythm or Ventricular Rate Control? AB - In patients with persistent atrial fibrillation, two therapeutic alternatives exist, namely restoration and maintenance of sinus rhythm versus ventricular rate control combined with anticoagulation. Currently, the selection of the best therapeutic strategy in an individual patient relies for the most part on clinical judgement and personal experience. At present, there are no prospective scientific data to support the superiority of one treatment over the other with respect to overall survival or quality of life. This review summarizes the present knowledge on this important clinical problem with particular emphasis on issues such as efficacy of antiarrhythmic drugs to prevent recurrent atrial fibrillation, proarrhythmic hazards of these compounds, or efficacy and safety of anticoagulation in nonrheumatic atrial fibrillation. These data serve as the basis of ongoing clinical trials prospectively comparing the merits and demerits of the two therapeutic strategies in the most common arrhythmia encountered in clinical practice. PMID- 10684497 TI - Atrial Fibrillation: Defining Some Unanswered Questions. PMID- 10684498 TI - Different Effects of Amiodarone and Quinidine on the Homogeneity of Myocardial Refractoriness in Patients With Intraventricular Conduction Delay. AB - BACKGROUND: Increases in QT and JT dispersion have been suggested as indicative of a proarrhythmic potential as a result of heterogeneity in myocardial refractoriness, the reduction of which by antiarrhythmic agents might be associated with a beneficial effect on the development of serious ventricular arrhythmias. METHODS: To test the hypothesis that amiodarone reduces the heter ogeneity of ventricular refractoriness to a significantly greater extent than quinidine in patients with intraventricular conduction defects under treatment for ventricular arrhythmias, the corrected and uncorrected QT and JT intervals and dispersions from 12-lead surface electrocardiograms were determined in 120 patients with intraventricular conduction defects with cardiac arrhythmias before and during treatment with amiodarone (n = 60) and quinidine (n = 60). RESULTS: Amiodarone increased QT from 403 +/- 50 ms to 459 +/- 47 ms (P <.001), with a similar increase in the corrected QT interval (QTc) (P <.001). Amiodarone reduced QT dispersion by 40% (P <.001), whereas quinidine increased by 18% (P <.001). The net effects of both drugs were similar for OTc. Amiodarone, but not quinidine, reduced heart rate significantly; amiodarone had no effect on the QRS; but quinidine increased if (P <.001). Quinidine as well as amiodarone increased the JT and JTc intervals significantly, but the effect of quinidine was qualitatively less striking. Amiodarone decreased the JT dispersion by 33% (P <.001) and JTc dispersion by 37% (P <.001). On the other hand, quinidine increased the corresponding values for JT and JTc by 18% (P <.001) and 21% (P <.001), respectively. The overall data on QT and JT dispersion indicate an improvement in the homogeneity of myocardial refractoriness with amiodarone treatment and the converse with quinidine treatment; this observation is consistent with a lower proarrhythmic propensity and mortality with amiodarone than with quinidine. Quinidine increased the QRS interval more than amiodarone, and the data indicate that in patients with intraventricular conduction defects, the monitoring of the JT interval might more accurately reflect changes in myocardial repolarization. CONCLUSIONS: Amiodarone and quinidine both increased the corrected and uncorrected QT and JT intervals; amiodarone decreased and quinidine increased the dispersion of these intervals, and these results suggested an improvement in the homogeneity of myocardial refractoriness as a result of amiodarone treatment and the converse as a result of quinidine treatment. Quinidine increased the QTS interval more than amiodarone, and the data indicate that in patients with intraventricular conduction defects, the monitoring of the JT interval might more accurately reflect changes in myocardial repolarization. PMID- 10684499 TI - Significance of Supraventricular Tachyarrhythmias After Coronary Artery Bypass Graft Surgery and Their Prevention by Low-Dose Sotalol: A Prospective Double Blind Randomized Placebo-Controlled Study. AB - BACKGROUND: The single most frequent complication after coronary artery bypass graft surgery is the occurrence of supraventricular tachyarrhythmias leading to a prolonged hospital stay. Although several drugs have been used to treat these arrhythmias, effective prevention was only possible with beta-blocking drugs in selected patients. It was, therefore, the aim of the present study to evaluate the significance of supraventricular tachyarrhythmias in presence of today's cardioprotective management in a broad spectrum of patients and to assess the possible preventive effect and safety of low-dose sotalol after coronary artery bypass graft surgery. METHODS AND RESULTS: In a prospective randomized double blind placebo-controlled trial, 220 consecutive patients referred for elective coronary artery bypass graft surgery were randomized to 80 mg sotalol twice daily (n = 110) or matching placebo (n = 110) for 3 months with the first dose given 2 hours before surgery. There were no significant differences in baseline characteristics between the two groups. Low-dose sotalol reduced the rate of supraventricular arrhythmias from 43% (placebo) to 25% (sotalol, P <.01), which was atrial fibrillation in 83%, flutter in 7%, and other supraventricular arrhythmias in 10%. Only 7% of all arrhythmias were observed after day 9. Hospital stay was 11 +/- 4 days in patients with supraventricular arrhythmias versus 9 +/- 2 days (P <.001) in patients without. On the fourth postoperative day, heart rate was lower in the sotalol group (75 +/- 12 versus 86 +/- 14 beats per min; P <.0001), but QTc was not significantly prolonged (sotalol, 0.44 +/- 0.03; placebo, 0.43 +/- 0.03; P, ns). Study medication had to be discontinued due to side effects in 6.4% of sotalol and 3.6% of placebo patients (P, ns), but relevant side effects occurred only in two sotalol patients late after surgery. CONCLUSIONS: These data show that without antiarrhythmic therapy the incidence of supraventricular arrhythmias after coronary artery bypass graft surgery is high (43%) and that supraventricular arrhythmias were associated with a prolonged hospital stay (+/-2 days). Prophylactic treatment with low-dose sotalol reduced the incidence of supraventricular arrhythmias significantly (by 40%), thereby reducing overall hospital stay in treated patients. Because more than 90% of all supraventricular arrhythmic episodes occurred within 10 days after surgery and considering the small proarrhythmic effect of sotalol late after surgery, prophylactic treatment with sotalol may be recommended for the first 10 postoperative days to safely reduce supraventricular tachyarrhythmias. PMID- 10684500 TI - Use of Lipid Drugs With Acute Myocardial Infarction Patients: An Examination of Physician Prescribing Behaviors. AB - BACKGROUND: Clinical trials have shown that the use of lipid-lowering agents in postmyocardial infarction (MI) patients reduces rates of subsequent coronary events, reduces coronary artery bypass surgery rates, and improves survival. Physician decisions to prescribe lipid-lowering drugs is influenced by a number of patient factors, including age, medical history, and serum cholesterol levels. The purpose of this study was to examine physician behaviors in prescribing lipid lowering therapy in patients after acute MI. METHODS AND RESULTS: A retrospective study was conducted at a local community-based hospital. A total of 129 patients with validated acute MI, hospitalized between January 1996 and December 1996, was included in the study sample. Variables abstracted included patient age, sex, race, primary diagnosis, medical history, lipid-lowering interventions of the discharge plan, and other discharge instructions regarding smoking cessation, activity, and dietary modification. Descriptive analysis was performed. The study showed that only 7 subjects (8.8%) were discharged on lipid-lowering drugs. Several patients who did not undergo therapy had either a low-density lipoprotein concentration of <130 (n = 13), a high-density lipoprotein concentration of >50 (n = 6), or were on hormone replacement therapy (n = 3). Dietary modification was advised in 100% of subjects (n = 54) for whom the data were included in the charts. CONCLUSIONS: The results of this descriptive study suggests that lipid lowering drugs are being utilized at low rates in the secondary prevention of acute MI. However, additional risk-lowering factors may play a role in the decision to discharge without drugs. Because of potential side effects associated with their use, a prudent path appears to be the norm in prescription of lipid lowering drug therapy for MI patients. PMID- 10684501 TI - Adrenomedullin Does Not Inhibit Human Platelet Aggregation. AB - BACKGROUND: Adrenomedullin (ADM) is a hypotensive peptide isolated from human pheochromocytoma extracts discovered in 1993 using an assay system designed to monitor its ability to increase rat platelet adenosine 3',5'-cyclic monophosphate (cAMP) levels. Physiological mediators that elevate cAMP levels, such as prostaglandin (PG)E(1) and PGI(2), have also been shown to inhibit platelet aggregation. Therefore, we have chosen to investigate the effect of ADM, a peptide shown to increase platelet cAMP levels, on human platelet aggregation. METHODS AND RESULTS: Platelet-rich plasma prepared from blood donors was incubated with ADM (10(-9)-10(-6) M) for 1 min at 37 degrees C before the addition of a submaximal dose of adenosine 5'-diphosphate (ADP). ADM did not alter the platelet aggregatory response to ADP. PGE(1), a substance known to inhibit ADP-induced platelet aggregation (10(-6) M), however, inhibited ADP induced platelet aggregation. In addition, the ADM induced a dose-dependent relation in rings of human chorionic arteries. CONCLUSIONS: These data may be interpreted to suggest that human platelets do not possess a functional ADM receptor couple with adenylate cyclase. PMID- 10684502 TI - A Multicenter Study Evaluating the Effects of Sevoflurane on Renal Function in Patients With Renal Insufficiency. AB - BACKGROUND: This multicenter study was undertaken to compare the effect of sevoflurane with that of isoflurane on renal function in 26 patients with pre existing renal insufficiency. Sevoflurane undergoes hepatic metabolism, with release of inorganic fluoride. Elevated fluoride levels have been associated with renal impairment in patients undergoing methoxyflurane anesthesia raising concerns about the nephrotoxic potential of sevoflurane. METHODS: Patients were ASA II or III class, with renal insufficiency defined by a preoperative serum creatinine concentration of 1.5-3.0 mg/dl. A standardized anesthetic regimen was used consisting of intravenous induction with propofol, vecuronium for muscle relaxation, and fentanyl for analgesia. Patients were randomized to receive either isoflurane or sevoflurane with 100% oxygen. Blood samples were obtained preoperatively and at 24, 48, and 72 h postoperatively for renal/electrolyte determinations. Blood samples for plasma fluoride measurement were obtained preoperatively. RESULTS: Plasma fluoride levels were significantly higher in patients receiving sevoflurane at all measurement points from 0 to 72 h postanesthesia. Mean peak fluoride concentration was 33.4 uM. The maximum fluoride value measured was 51.2 uM. There were no significant differences in postoperative serum creatinine values at any time between patients receiving sevoflurane or isoflurane. CONCLUSIONS: Sevoflurane metabolism produces elevations in plasma fluoride concentrations relative to isoflurane. Despite the increase in plasma fluoride levels, the administration of sevoflurane to patients with renal insufficiency did not produce any adverse effects on renal function as measured by serum creatinine concentration when compared with isoflurane. PMID- 10684503 TI - Pharmacokinetics of Catecholamines During Hemofiltration in Pediatric Patients. AB - BACKGROUND: Continuous hemofiltration is used in pediatric patients with acute renal failure. Many of these patients are treated with catecholamines for hemodynamic instability. The authors studied the pharmacokinetics of dopamine and dobutamine on patients undergoing continuous venovenous hemofiltration. METHODS AND RESULTS: Three critically ill pediatric patients with acute renal failure and cardiovascular instability treated with hemofiltration and intravenous infusion of dopamine and/or dobutamine were entered into the study. Blood samples were drawn at steady-state levels from the arterial port (inflow) of the hemofilter, from the venous port (outflow) of the hemofilter, and from the ultrafilter. Sixteen (n = 16) pharmacokinetic measurements were made, six (n = 6) for dopamine and 10 (n = 10) for dobutamine. The clearance of dopamine by the hemofilter was 0.078 +/- 0.011 mL/kg/min, and for dobutamine it was 0.036 +/- 0.008 mL/kg/min. On the average, 0.56% of the dopamine dose and 0.13% of the dobutamine dose was removed by the hemofilter. CONCLUSIONS: The pharmacokinetics of dopamine and dobutamine at the dosage range of 5-25 ug/kg/min are not altered by continuous hemofiltration. Relative to total plasma clearance, negligible amounts of the drug were removed. The dosage of these catecholamines need not be adjusted during continuous hemofiltration. PMID- 10684504 TI - Effect of beta-Adrenoceptor Antagonists on Phospholipid N-Methylation Activities of Cardiac Sarcolemma. AB - BACKGROUND: Some beta-adrenoceptor antagonists exert a negative inotropic action by affecting Ca(2+) fluxes in the myocardial cell as a consequence of their interaction with sarcolemmal and sarcoplasmic reticular membranes. This action may be caused by their effects on the chemicophysical properties of membranes phospholipids. Because phosphatidylethanolamine (PE) N-methylation can influence the chemicophysical properties of membranes, these agents may affect PE N methylation. This study was undertaken to examine the effects of propranolol, acebutolol, and atenolol on PE-N-methylation in rat heart sarcolemma (SL). METHODS AND RESULTS: Sarcolemmal membrane was isolated from rat hearts by the hypotonic shock LiBr method. Incorporation of radiolabeled methyl groups from S adenosyl-l-methionine was assayed at three catalytic sites involved in the PE N methylation reaction in the presence and absence of these drugs. A biphasic effect of propranolol at site I was noted; low concentrations (10(-8) M) were inhibitor. Acebutolol (10(-9)-10(-3) M) depressed methyl group incorporation in SL at site II in a dose-dependent manner, whereas atenolol showed no effect. Propranolol also exerted a biphasic effect on sarcoplasmic reticular (SR) methylation at site I, whereas acebutolol depressed the SR enzyme activity at site II and atenolol had no effect. The mitochondrial methyltransferase activities at sites I, II, and III were unaltered by any of these drugs. CONCLUSIONS: It is suggested that propranolol and acebutolol alter SL and SR PE N methyltransferase activity at site I and site II, respectively, either by affecting the enzyme directly or by changing the physiochemical properties of the membrane. PMID- 10684505 TI - Analysis of Vasodepressor Responses to Nociceptin and Nociceptin Analogs in the Systemic Vascular Bed of the Anesthetized Rabbit In Vivo. AB - Background: The heptadecapeptide nociceptin, also known as Orphanin FQ, is a recently discovered endogenous ligand for the opioid-like G-protein-coupled receptor ORL(1). Methods and Results: In the present study, responses to nociceptin, [Tyr(1)]-nociceptin, nociceptin-(2-17), nociceptin-(1-11), and nociceptin-(1-7) were compared in the systemic vascular bed of the rabbit. Nociceptin and [Tyr(1)]-nociceptin induced dose related decreases in systemic arterial pressure (SAP) when injected in doses of 1-30 nmol/kg intravenous (IV); in terms of relative vasodepressor activity, [Tyr(1)]-nociceptin and nocicpetin were similar in potency. However, nocicpetin-(2-17), nocicpetin-(1-11), and nociceptin-(1-7) had no effect on SAP when injected in doses up to 30 nmol/kg IV. The decreases in SAP in response to nociceptin and [Tyr(1)]-nociceptin were not altered by the opioid receptor antagonist naloxone at a time when depressor responses to methionine-enkephalin were reduced significantly. Conclusions: The results of the present study show that vasodepressor responses to nociceptin and [Tyr(1)]-nocicpetin are mediated by the activation of a naloxene-insensitive opioid receptor and are not dependent on the presence of Phe at the N-terminus of the nociceptin sequence. Moreover, the present results show that nociceptin-(2 17), nociceptin-(1-11), and nociceptin-(1-7) do not alter SAP in the rabbit, indicating that peptide chain length is important for the expression of vasodepressor activity. PMID- 10684506 TI - Nociceptin Has Vasodilator Activity in the Pulmonary Vascular Bed of the Rat. AB - BACKGROUND: Recent studies have shown that the newly discovered endogenous opioid like peptide, nociceptin, has vasodilator activity in the peripheral vascular bed. However, little if anything is known about the effects of the peptide in the pulmonary vascular bed. Therefore, responses to nociceptin in the pulmonary vascular bed were investigated and compared with responses in the hindlimb and systemic vascular beds of the rat. METHODS AND RESULTS: Responses to nociceptin were investigated in the pulmonary and hindquarters vascular beds in the rat under constant flow conditions and were compared with decreases in systemic vascular resistance. Under conditions of constant flow, injections of nociceptin in doses of 3-30 nM induced dose-related decreases in pulmonary arterial perfusion pressure when baseline tone was increased to a high steady level with U46619. Pulmonary vasodilator responses to nociceptin were not modified by the opioid receptor antagonist naloxone, and the newly discovered ligand was 10-fold less potent than adrenomedullin in decreasing pulmonary vascular resistance when decreases in pulmonary hind quarters and systemic vascular resistances were compared, nociceptin was significantly more potent in decreasing hindquarters and systemic vascular resistance than in reducing pulmonary vascular resistance. CONCLUSIONS: The results of the present study indicate that nociceptin decreases pulmonary vascular resistance by a naloxone-insensitive mechanism, and that the peptide is less potent in decreasing pulmonary vascular resistance than in decreasing systemic vascular resistance in the rat. PMID- 10684507 TI - Significance and Prevention of Atrial Fibrillation Occurring After Cardiac Surgery: A Time for Fundamental Change in Strategy? PMID- 10684508 TI - Risk Factors for Tachycardia Events Caused by Antiarrhythmic Drugs: Experience From the ESVEM Trial. AB - BACKGROUND: In the Electrophysiology Study versus Electrocardiographic Monitoring (ESVEM) trial, up to seven antiarrhythmic drugs were randomly assigned to 486 patients with a history of sustained ventricular arrhythmia. At baseline, all the patients had inducible sustained ventricular tachycardia (VT) and had >/=10 premature ventricular beats (PVBs) per hour on 48-hour Holter monitoring. A total of 1,229 drug trials were performed. Antiarrhythmic drugs were discontinued during hospitalization because of ventricular tachyarrhythmias thought to be a proarrhythmic effect of the antiarrhythmic drugs in 96 of 479 patients (20%) who received drugs. Proarrhythmic effects were defined as sustained VT, ventricular fibrillation or arrhythmic death, torsade de pointes, or distinct intolerable worsening of the baseline arrhythmia after at least three doses of the drug. METHODS AND RESULTS: Eighteen baseline characteristics were analyzed for factors that would predict a higher incidence of proarrhythmia. These included type of heart disease, previous myocardial infarction, symptom activity scale, gender, type of arrhythmia, VT/ventricular fibrillation, age, left ventricular ejection fraction (LVEF), PVB frequency, heart rate, QRS duration, and QT interval. Multiple logistic regression analysis identified increased mean PVB frequency (P =.003) and increased heart rate (P =.026) as significant predictors of proarrhythmia. Decreased LVEF (<25%) exhibited only a trend toward significance (P =.073). When proarrhythmia was redefined as sustained VT, cardiac arrest of arrhythmic death, or torsade de pointes (n = 59), PVB frequency (P =.003) and heart rate (P =.034) were still the only significant baseline predictors. CONCLUSIONS: In the ESVEM study, higher PVB frequency and higher heart rate were significant predictors of drug-induced proarrhythmia. PMID- 10684509 TI - Efficacy of Nicardipine for Long-Term Therapy of Chronic Stable Angina. AB - BACKGROUND: Although angina is a chronic disease, most clinical trials evaluating antianginal therapy are of a few weeks or months in duration. METHODS AND RESULTS: To evaluate the effects of nicardipine, a second-generation dihydro pyridine calcium channel blocker, as long-term therapy, patients with chronic stable angina were treated for 1 year with open-label nicardipine. Anginal symptoms were controlled with 20 mg tid in 24%, 30 mg tid in 34%, and 40 mg tid in 42%. Of 72 patients completing the 1-year trial, only 14 (19%) had required the addition of long-acting nitrates for control of symptoms. The remaining 57 patients, who had anginal symptoms controlled with nicardipine alone, were continued into the 3-week, double-blind period and were randomized to continue their usual dose of nicardipine or placebo. Throughout the 1-year, open-label treatment period, the number of anginal episodes and exercise parameters of angina were significantly reduced with nicardipine. CONCLUSIONS: During the double-blind period, the patients who continued on nicardipine had significantly greater exercise time and time to onset of angina than patients who were randomized to placebo. The exercise parameters in the patients randomized to placebo were improved over baseline after 1 year of therapy; however, the improvement with nicardipine was significantly greater. PMID- 10684510 TI - Effect of Digoxin on Ventricular Remodeling and Responsiveness of beta Adrenoceptors in Chronic Volume Overload. AB - BACKGROUND: Digoxin improves baroreflex function and reduces neurohumoral activation in severe heart failure, but it is uncertain how digoxin affects ventricular remodeling and progression to left ventricular dysfunction. In addition, the effect of digoxin in in vitro beta-adrenoceptor density and function, and contractile reserve in vivo is not well understood. METHODS AND RESULTS: To study this, we compared digoxin with placebo treatment in rats with chronic volume overload induced by aortocaval fistula and in sham-operated control animals. Left ventricular end-diastolic cavity dimensions (LVDd) and wall thickness were measured weekly by in vivo transthoracic echocardiography, and left ventricular mass (LVM) and percent fractional shortening (%FS) were calculated. Six weeks after fistula creation, simultaneous echocardiographic and invasive hemodynamic evaluation at rest and in response to incremental dobutamine (1-10 ug/kg/min intravenously) were measured. Myocardial plasma membrane beta adrenoceptor density and maximal adenylate cyclase responses (V(max)) to isoproterenol, 5'-guanylylimi dodiphosphate, and forskolin were measured in vitro. Volume overload induced progressive increases in LVDd and LVM over the 6 week study period. Percent fractional shortening at rest, and the change in %FS in response to dobutamine stress were dramatically reduced 6 weeks after fistula creation. Although 6-week fistula animals had unchanged beta-adrenoceptor density (B(max)) and binding affinity (K(d)) as compared with controls, maximal adenylate cyclase responses to stimulation in vitro (V(max)) were markedly reduced. Digoxin treatment prevented this loss of responsiveness of adenylate cyclase but did not affect beta-adrenoceptor density or affinity in vitro. Digoxin had no effect on LVDd, LVM, %FS, or the response to dobutamine infusion in vivo. CONCLUSIONS: Although digoxin prevented beta-adrenoceptor desensitization and improved in vitro myocardial adenylate cyclase response, the cardiac response to adrenergic stimulation in vivo was not significantly improved. These results suggest that the role of beta-adrenoceptor desensitization in the progression from volume overload hypertrophy to left ventricular dysfunction and heart failure may be less important than previously thought. Furthermore, although digoxin treatment did produce modest hemodynamic benefits, it did not prevent progressive remodeling in this model. PMID- 10684511 TI - Modification of the ATP-Induced Increase in AB - BACKGROUND: It is generally accepted that the plasma membrane of mammalian ventricular myocytes regulates the cytosolic concentration of Ca(2+). In this study we investigated the effects of some P2-purinoceptor antagonists and metals such as copper and zinc on the adenosine triphosphate (ATP)-induced increase in intracellular concentration of free Ca(2+) ([Ca(2+)](i)). METHODS AND RESULTS: Cardiomyocytes were isolated from adult male Sprague-Dawley rats loaded with Fura 2, and fluorescence measurements were performed by employing stirred cell suspensions at room temperature. ATP (50 uM) increased [Ca(2+)](i) over the basal value, and 10 uM cibacron blue or verapamil virtually abolished it. The ATP induced increase in [Ca(2+)](i) was not observed in Ca(2+)- or Mg(2+)-free buffers. Incubation of cells with ZnCl(2) produced a significant depression of the ATP-induced increase in [Ca(2+)](i); 25 uM Zn(2+) decreased the peak response to approximately 50% of the control value. The ATP-induced increase in [Ca(2+)](i), was inhibited by low concentrations (1-5 uM) of Cu(2+) but was markedly augmented by high concentrations (25 uM) of Cu(2+). The increase in the [Ca(2+)](i) response to cron blue, and Zn(2+), but not by ryanodine or caffeine pretreatment. CONCLUSIONS: The ATP-induced increase in [Ca(2+)](i) is dependent on the extracellular concentrations of Ca(2+) as well as Mg(2+) and is antagonized by cibacron blue and Zn(2+). On the other hand, Cu(2+) produced a biphasic response to the ATP-induced increase in [Ca(2+)](i) in cardiomyocytes. PMID- 10684512 TI - Effects of Intracellular Calcium Reduction by Dantrolene on Prevention/Treatment of Ischemic Stroke. AB - BACKGROUND: It has been suggested that cerebral blood vessels and brain cells might depend more on intracellular calcium than extracellular calcium to modulate intracellular free calcium concentrations, [Ca(2+)](i). METHODS AND RESULTS: A potent intracellular calcium antagonist, dantrolene, was used to prevent the ischemic stroke induced in the rat model. It was found that treatment of rats with dantrolene at -1 hour and +1 hour after 60 minutes of ischemic insult prevented by the formation of cortical necrosis 98% and 85%, respectively. Further, the [Ca(2+)](i) of embryonic aorta cells was markedly reduced, and cAMP of the same cultured cells were significantly increased by dantrolene treatment. CONCLUSIONS: These results indicate that ischemic stroke is preventable by dantrolene through reduction of [Ca(2+)](i) and increase of cAMP. PMID- 10684513 TI - Oxygen Free Radicals and Endotoxic Shock: Effect of Flaxseed. AB - BACKGROUND: Cardiac dysfunction and tissue injury during endotoxemia may be caused by increased levels of oxygen free radicals. METHODS AND RESULTS: We therefore investigated the effects of endotoxic shock on cardiac function and contractility, plasma creatine kinase (CK) activity and lactate concentration, oxyradical-producing activity of polymorphonuclear leukocytes (PMNL-CL) and white blood corpuscles, antioxidant reserve (cardiac chemiluminescence [LV-CL]), antioxidant enzyme activity (superoxide dismutase, catalase, glutathione peroxidase), cardiac malondialdehyde (MDA) concentration, a lipid peroxidation product, and hemodynamics in the absence or presence of flaxseed treatment in anesthetized dogs. Flaxseed contains lignans that have antioxidant activites and inhibit platelet-activating factor (PAF). The dogs were assigned to three groups: group I, sham control; group II, endotoxin (ET) treated (5 mg/kg intravenously); group III, ET + flaxseed (2 gm/kg/day orally) for 6 days. ET produced a decrease in cardiac function and contractility and antioxidant enzyme levels, and an increase in cardiac MDA and LV-CL, PMNL-CL, and plasma CK and lactate. Pretreatment with flaxseed attenuated the ET-induced cardiac dysfunction and cellular damage. Protection was incomplete for cardiovascular function, plasma CK, and lactate. CONCLUSIONS: These results suggest that oxyradicals and/or PAF may be involved in the deterioration of cardiovascular function and cellular integrity during ET shock and that antioxidant and anti-PAF agents may be effective in the treatment of ET shock. PMID- 10684514 TI - Effects of Metformin on Collagen Glycation and Diastolic Dysfunction in Diabetic Myocardium. AB - BACKGROUND: Collagen accumulation in the myocardial interstitium of diabetic animals is considered to promote diastolic stiffness through advanced glycosylation. Because in vitro data suggest that metformin can modify glycosylation, this study was undertaken in a canine diabetic model 4 months in duration. METHODS AND RESULTS: Untreated diabetics (group II) and diabetics treated with metformin alone (group III) or with insulin (group IV) were compared in the basal state and during volume infusion. Basal hemoglobin A(1c), heart rate, aortic pressure, and ejection fraction were comparable. Left ventricular end-diastolic pressure was significantly increased in the untreated diabetics of group II, associated with a reduced end-diastolic volume. By contrast these parameters in the metformin-treated diabetics of group III were comparable with those in the normals of group I. Similarly in group IV end-diastolic volume was higher than that in group II, but filling pressure, although lower, was not significantly so. Calculation of left ventricular chamber stiffness in the basal state indicated a higher level for group II compared with controls and the treatment groups. During the systemic infusion of dextran, the untreated diabetics of group II had the largest end-diastolic pressure increase and the smallest rise of end-diastolic volume of the treatment groups, consistent with a significantly greater chamber stiffness. Myocardial collagen concentration was increased in group II with an interstitial distribution on morphological exam. Levels of collagen-linked advanced glycosylation end products isolated from the left ventricular were significantly greater in group II than in group I. Treatment with metformin prevented the increment observed in the untreated diabetic but had no effect on the elevated collagen concentration. CONCLUSIONS: Untreated diabetics exhibited increased diastolic chamber stiffness associated with collagen-linked glycation in myocardium compared with control animals. Chronic metformin use prevented the abnormalities of function and composition. PMID- 10684516 TI - Controlling Cardiac Arrhythmias: New Drugs in Development and Insights From Molecular Biology. PMID- 10684515 TI - Angiotensin-Converting Enzyme Inhibition Delays Onset of Glucosuria With Regression of Renal Injuries in Genetic Rat Model of Non-Insulin-Dependent Diabetes Mellitus. AB - BACKGROUND: The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is a new genetic model of non-insulin-dependent diabetes mellitus (NIDDM). We investigated whether angiotensin inhibition influences the onset of NIDDM and brings about a regression of renal injury in diabetes mellitus. METHODS AND RESULTS: Six-week old OLETF rats were treated with the angiotensin-converting enzyme (ACE) inhibitors imidapril or enalapril for 16 weeks. Systolic blood pressure is increased in an age-dependent manner in OLETF rats. In this study, the elevation in systolic blood pressure was dose-dependently reduced by ACE inhibitor treatment. In OLETF rats, plasma concentrations of insulin and glucose increased and the glucosuria occurred at the age of 22 weeks. Simultaneously, OLETF rats exhibited proteinuria and nodular lesions in glomeruli. The ACE inhibitor treatment almost completely reduced glucosuria, and also decreased plasma concentrations of insulin and glucose in OLETF rats. ACE inhibitor treatment lessened the proteinuria and attenuated morphologically the severity of nodular lesions in OLETF rats. Moreover, increases in plasminogen activator inhibitor 1 (PAI-1) in OLETF rats were reduced by the ACE inhibitor treatment, and the improvement of glomerular lesions was related to decreases of PAI-1 and angiotensin II levels in plasma but not to improvement of glucose metabolism. CONCLUSIONS: ACE inhibitors delay onset of NIDDM with attenuation of kidney injury. The regression of kidney lesions is probably due to angiotensin reductions but not to glucose metabolism per se. ACE inhibitor drug therapy may be useful in preventing NIDDM and the subsequent renal injury in patients with NIDDM. PMID- 10684517 TI - Editorial Page. PMID- 10684518 TI - Initiating and Maintaining Patients on Warfarin Anticoagulation: The Importance of Monitoring. AB - BACKGROUND: The VA Stroke Prevention in Nonrheumatic Atrial Fibrillation study was a prospective, randomized, double-blind study comparing low-dose warfarin with placebo in patients with nonrheumatic atrial fibrillation. The trial showed a 79% reduction in stroke rate in warfarin randomized patients without an increase in bleeding complications. We examined the need for frequent prothrombin time monitoring (international normalized ratios [INR] were not measured directly) in patients receiving warfarin.

.05). Irbesartan significantly lowered BP without clinically important changes in renal function. Irbesartan had no effect on 24-hour urinary TXA(2)-M excretion, but significantly increased 24-hour PGI(2)-M excretion versus placebo on day B29 (20.7 +/- 23 pg/mg creatinine vs _2.3 +/- 43 pg/mg creatinine; P <.05). Pharmacokinetics were comparable to those from previous studies. The hourly relationship between plasma irbesartan concentration and antihypertensive effect indicated a broad, clockwise hysteresis, with peak concentration occurring at 1.5 hours, whereas peak antihypertensive effect occurred at 4 hours. CONCLUSIONS: Irbesartan increases plasma AII and PRA and lowers BP consistent with AT(1) receptor blockade, without clinically important effects on renal function. PMID- 10684526 TI - Prevention of Vascular Apoptosis in Myocardial Infarction by Losartan. AB - BACKGROUND: Previous studies have demonstrated the occurrence of apoptosis in cardiomyocytes in different types of cardiovascular diseases. This report provides the first evidence for the presence of vascular apoptosis in myocardial infarction induced in rats by occluding the coronary artery for 7 weeks. METHODS AND RESULTS: Apoptosis was characterized by DNA fragmentation, upregulation of caspase-3, downregulation of poly (ADP-ribose) polymerase (PARP), increased c-fos mRNA expression and caspase-3/PARP ratio in aortic vascular smooth muscle cells. The results show apoptotic changes in 10-25% of the aortic vascular cells after myocardial infarction; these alterations were prevented after treating the 3-week operated animals with an angiotensin II receptor antagonist, losartan (25 mg/kg/day; intraperitoneal) for 4 weeks. Cultured rat aortic smooth muscle cells exposed to 10 nmol/L angiotensin II for 48 hours also exhibited apoptotic changes, which were inhibited by 10 nmol/L losartan. CONCLUSIONS: These results suggest that vascular apoptosis occurs in myocardial infarction, and this may be due to an increase in the circulating levels of angiotensin II. PMID- 10684527 TI - Effects of Palmitoyl Carnitine on Perfused Heart and Papillary Muscle. AB - BACKGROUND: Palmitoyl carnitine accumulation during ischemia causes profound electrophysiological changes, resulting in arrhythmias. We studied the electrophysiological and contractile effects of palmitoyl carnitine. METHODS AND RESULTS: Extracellular recordings made by using the endocardial unipolar paced evoked response (PER) in isolated perfused rabbit hearts were compared with action potentials (AP) recorded from septal artery perfused rabbit papillary muscle. Left ventricular pressure was monitored in isolated hearts. In perfused hearts palmitoyl carnitine (30 umol/L, 30 minutes) significantly (P <.001) increased the latency of activation (St-R interval) by 58% +/- 8% and reduced repolarization time (R-E interval) by 39% +/- 4%. PER duration (St-E interval), was reduced by 30% +/- 3%. Palmitoyl carnitine (30 umol/L) significantly (P <.001) decreased resting membrane potential (19 +/- 2 mV) of AP, reduced peak amplitude (33.5 +/- 8 mV) and rate of rise of phase 0 (41 +/- 8 V/s). Significant reductions (P <.001) in the action potential duration 50% (129.4 +/- 28 ms) and 90% (139.8 +/- 32 ms) were also observed. An initial positive inotropic effect, which declined as irreversible contracture developed, was also observed. Verapamil (1 umol/L), nifedipine (1 umol/L), and caffeine (10 mmol/L) failed to abolish the positive inotropy. CONCLUSIONS: We suggest that palmitoyl carnitine disrupts intracellular calcium homeostasis leading to disturbances in electrical and contractile activity. Its accumulation during myocardial ischemia could contribute to calcium overloading and initiate lethal arrhythmias. PMID- 10684528 TI - The Effect of Chloroquine on Circulating Platelet Survival in the Rat. AB - BACKGROUND: Chloroquine inhibits platelet aggregation. Because platelet aggregation may lead to the lysis of platelets, the effect of chloroquine administration on circulating platelet survival was studied. METHODS AND RESULTS: Platelets harvested from the blood of male inbred (WAG) rats were labeled with (111)Indium oxine and returned to other inbred rats. At timed intervals, blood samples were drawn from the rats and taken for radioactivity estimation. In some experiments, the rats (n = 10) received chloroquine (10 mg/kg) intraperitoneally daily for 3 days. Control rats (n = 10) received chloroquine (10 mg/kg) intraperitoneally daily for 3 days. Control rats (n = 10) received normal saline. Indium-labeled platelets disappeared exponentially in control and test rats. The fraction of indium disappearing/h was significantly less in chloroquine-treated rats than in control rats (0.0368 +/- 0.0016 vs 0.0520 +/- 0.0016, mean +/- SEM; P <.001). In all, 99% of the labeled platelets disappeared in 5.3 days in chloroquine-treated rats and 3.7 days in control rats. CONCLUSIONS: Chloroquine administration increases the life span of circulating platelets in rats. If the results are confirmed in humans, chloroquine may prevent the shortened platelet survival and thrombocytopenia common in malarial infection and other thrombotic disorders. PMID- 10684529 TI - Increased Intramural Retention After Local Delivery of Molecules with Increased Binding Properties: Implications for Regional Delivery of Pharmacologic Agents. AB - BACKGROUND: Catheter-based local vascular delivery results in concentrated qualtities of pharmaceutical agents or genes into focal areas of the arterial wall. However, intramural retention is short and has reduced the potential efficacy of this approach. It was postulated that agents that possess increased intramural binding would show increased intramural retention. Platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) were models of agents with increased cellular and extracellular matrix binding properties. METHODS AND RESULTS: The delivery efficiency and intramural retention of 2 mL of saline containing I(125) labeled PDGF (n = 35 arteries) and bFGF (n = 24) were compared with albumin (n = 21) after local delivery into porcine coronary arteries. Animals were sacrificed at three or more prespecified timepoints: immediately after delivery, 1 day, or 3 days after delivery and if necessary at 5 or 7 days to document prolonged retention. Autoradiograms of the arterial sections were evaluated for the extent of delivery. Delivery efficiency, defined as the amount leaving the catheter and retrieved from the arterial wall, was 0.60% +/- 0.42% for albumin, 1.98% +/- 0.88% for PDGF (P =.001), and 0.31% +/- 0.11% for bFGF. The calculated intramural half-life of albumin was 7.4 hours, 56.2 hours for PDGF, and 14.9 hours for bFGF (P =.0001 for PDGF). Infusate covering >50% of the medial area was observed in 85% of arteries immediately after delivery. Although myocardial delivery was similar for albumin, PDGF, and bFGF, myocardial retention was significantlylonger for bFGF (P <.001). CONCLUSIONS: Molecules that exhibit preferential intramural binding show a longer intramural residence duration than solutes without such binding properties. In addition, delivery and subsequent prolonged retention in the myocardium can be obtained by local delivery via the arterial lumen of solutions with preferential binding properties. PMID- 10684530 TI - Persistence of Atrial Fibrillation After Its Induction-Importance of the Duration and Dispersion of Atrial Refractoriness and Electrical Remodeling. AB - BACKGROUND: The electrophysiologic mechanisms of the persistence of atrial fibrillation (AF) after its initiation are not well understood. Therefore, the electrophysiologic characteristics of the right atrium were evaluated in an acute, pacing-induced model of AF in the pig in order to identify parameters associated with persistence of AF. METHODS AND RESULTS: AF was induced by rapid atrial pacing in 30 anesthetized, open-chest, juvenile pigs. Sustained (S) AF was defined as that lasting >10 minutes, nonsustained (NS) AF <10 minutes but >30 seconds, and no (N) AF <30 seconds. Activation mapping and programmed stimulation (S1S1 = 200 ms) was performed at 56 electrodes on the right atrial free wall, to determine ERP (mean and minimum), dispersion of refractoriness (ERPdisp, ELEdisp), conduction velocity (CV), wavelength, AF cycle length (mean of 10 beats), and AF cycle length/time (electrical remodeling). SAF was induced in 10 pigs, NSAF in 9, and NAF in 11. AF cycle length was shorter in SAF and/vs NS vs NAF (P <.001). Mean ERP (107 +/- 9 and/vs 122 +/- 5 vs 142 +/- 9, p <.001) and wavelength (7 +/- 1 and/vs 9 +/- 1 vs 11 +/- 1, P <.001) were shorter in SAF and/vs NSAF vs NAF. Minimum ERP was shorter in SAF and NSAF vs NAF (P <.001). CV at cycle lengths of 200 and 150 msec was not different between groups. Dispersion of ERP was greater in SAF and/vs NSAF vs NAF (8 +/- 1 and/vs 11 +/- 1 vs 19 +/- 4, P <.001). CONCLUSIONS: Persistence of AF correlated with shorter ERP and wavelength, and greater dispersion of ERP and electrical remodeling. There was no correlation with CV. PMID- 10684531 TI - Nicotine Increases Spatiotemporal Complexity of Ventricular Fibrillation Wavefront on the Epicardial Border Zone of Healed Canine Infarcts. AB - BACKGROUND: The influence of a pharmacologic agent on wavefront dynamics during ventricular fibrillation (VF) in a setting of remodeled and healed myocardial infarction (MI) remains poor explored. We hypothesized that nicotine, by virtue of its complex direct and indirect cardiovascular effects, increases wavefront complexity during VF. Specifically, we sought to determine whether nicotine increases the number and complexity (approximate entropy) of wavelets during stage II VF in hearts with healed MI. METHODS AND RESULTS: The left anterior descending coronary artery was permanently occluded in five mongrel dogs and wavefront dynamics during VF studied 5 to 6 weeks after occlusion in the open chest anesthetized state. VF was induced by rapid pacing and the activation pattern mapped on the surviving epicardial border zone (EBZ) of the left ventricle with a plaque (3.2 x 3.8 cm) having 477 bipolar electrodes 1.6 mm apart. VF was mapped before and 20 minutes after 5 ug/kg/min nicotine infusion. Nicotine with a mean arterial plasma concentration of 127 +/- 76 ng/mL (range 57 to 240 ng/mL) significantly (P <.01) increased the number of wavelents from 3.8 +/- 0.4 to 5 +/- 0.41. The increased number of wavelets was caused by an increase (P <.01) in the spontaneous breakup of wavefronts from 4.1 +/- 0.9 times/s to 6.9 +/- 1.1 times/s. Wavebreak over the EBZ was functional in nature as no breakup occurred during normal sinus rhythm. Approximate entropy, a measure of complexity, significantly (P <.01) increased after nicotine administration from 0.23 +/- 0.02 to 0.28 +/- 0.01. CONCLUSIONS: Nicotine increases the number of wavelets and their complexity during VF by promoting spontaneous wavebreak over the EBZ of healed MI. PMID- 10684532 TI - Recurrent Torsades de Pointes After Sotalol Therapy for Symptomatic Paroxysmal Atrial Fibrillation in a Patient with End-Stage Renal Disease. AB - BACKGROUND: In recent years there has been ain increase in the use of class III antiarrhythmic drugs such as sotalol, amiodarone, and the so-called pure class III compound for the control of cardiac arrhythmias. It appears there has been a corresponding increase in the frequency of torsades de pointes (TdP). METHODS AND RESULTS: The case reported here, a patient on daily renal dialysis for end-stage renal disease, has important implications for class III agents, which are excreted largely by the kidneys. A relatively low dose of sotalol administered for the prevention of recurrences of atrial fabrillation, with a fast ventricular response producing angina, led to modest increases in the QT interval and moderate bradycardia. This culminated in the development of TdP, which deteriorated into ventricular fibrillation, from which the patient could be resuscitated with considerable difficulty. Dialysis after the occurrence of TdP led to further and striking prolongation of the QT interval associated with numerous episodes of TdP for several days before control was achieved. The atrial fibrillation and recurrences of TdP were eventually controlled with oral amiodarone. CONCLUSIONS: This case emphasizes that in the absence of significant renal function, use of sotalol may not be safe because drug accumulation may not be controlled adequately with renal dialysis. In view of this, in patients with end-stage renal disease, the use of sotalol for arrhythmia control appears contraindicated and alternative agents, the excretion of which does not occur by the renal route, should be used. PMID- 10684533 TI - Increase in Plasma Angiotensin II Levels After Chemical Blockade of AT1 Receptor Blockers, But Not With Antisense Oligodeoxynucleotide Directed at AT1 Receptor mRNA: A Major Benefit of Gene Therapy. PMID- 10684534 TI - Protamine Reversal of Heparin After Cardiopulmonary Bypass Increases Lung Resistance, Not Elastance. AB - BACKGROUND: Protamine, an immunologically active, cationic amine, has been suspected of impairing lung mechanics when administered after cardiopulmonary bypass (CPB) to reverse heparin. Whether such adverse changes are an effect of protamine itself, the formation of heparin-protamine complexes, the extent of heparin anticoagulation, or its chemical reversal is not known. METHODS AND RESULTS: Using a computer-controlled, forced-ventilation method over a variety of physiological tidal volume (V(T)) and frequency (f) combinations, we prospectively studied 18 adult, elective patients before systemic heparinization and after protamine reversal to confirm and, possibly, elucidate an etiology for any adverse pulmonary effects. Protamine and heparin doses, their sum (Sigma dose) and differential (Delta-dose) doses, and activated clotting times were tabulated. In all patients, lung resistance (R(L)) and, to a lesser extent, elastance (E(L)) increased after CPB, compared with pre-CPB values (P <.05). However, R(L) particularly increased after CPB with increases correlated to the Delta-dose, where R(LPRE-->POST) = -0.037 [Delta-dose] - 0.56f ?_ 0.019V(T) + 36.1 (r =.652, P <.05). No other significant correlations were found among the remaining clinical parameters and changes in either R(L) or E(L), or any chest wall component (all P >.05). CONCLUSIONS: The changes seen in R(L) after CPB were greatest in those patients receiving the most nearly balanced doses of heparin and protamine, and were not related significantly to the total heparin or protamine doses, or their sum. These suggests that the extent of anticoagulation reversal or formation of heparin-protamine complexes, and not protamine itself, are more responsible for changes seen in lung mechanics. The changes seen were limited solely to R(L), and not in either E(L) nor the chest wall mechanical properties. PMID- 10684535 TI - Concomitant Block of the Rapid (I(Kr)) and Slow (I(Ks)) Components of the Delayed Rectifier Potassium Current is Associated With Additional Drug Effects on Lengthening of Cardiac Repolarization. AB - BACKGROUND: The delayed rectifier potassium current, which comprises both a rapid (I(Kr)) and as slow (I(Ks)) component, is a major outward current involved in repolarization of cardiac myocytes. I(Kr) is the target of most drugs that prolong repolarization, whereas electrophysiological effects resulting from combined block of I(Kr) and I(Ks) still need to be characterized. METHODS AND RESULTS: Studies in isolated, buffer-perfused guinea pig hearts were undertaken to compare lengthening of cardiac repolarization under conditions of I(Kr) block alone, I(Ks) Block alone, or combined block of I(Kr) and I(Ks). In protocol A, isolated perfusion with N-acetylprocainamide (NAPA) (I(Kr) block), indapamide (I(Ks) block), or combined NAPA/indapamide was performed at a pacing cycle length of 250 msec. Increases in monophasic action potential duration measured at 90% polarization (MAPD(90)) from baseline after perfusion with NAPA 100 umol/L (IC(50) for block of I(Kr)) was 19 +/- 6 msed (P <.05), after indapamide 100 umol/L (EC(50) for block of I(Ks)) 13 +/- 2 msec (P <.05), but 42 +/- 5 msec after combined NAPA 100 umol/L and indapamide 100 umol/L (P <.05 vs. baseline and isolated administrations), suggesting the possibility of excessive lengthening of cardiac repolarization by blocking both I(Kr) and I(Ks). As well, in protocol B where sequential perfusions with dofetilide (I(Kr) blocker), dofetilide/indapamide, and indapamide in the same hearts were used, combined dofetilide/indapamide infusion showed a greater increase in MAPD(90) during all pacing cycles studied (250 to 150 msec). CONCLUSIONS: Combined I(Kr) and I(Ks) block may lead to excessive lengthening of cardiac repolarization. This may predispose patients to proarrhythmia during coadministration of drugs. PMID- 10684536 TI - The Effect of Agmatine on Ischemic and Nonischemic Isolated Rat Heart. AB - OBJECTIVE: the natural polyamines play a protective role during ischemic injury. We studied the effects of agmatine on ischemic and nonischemic isolated rat hearts. METHODS: Thirty-one rats were randomly assigned to one of four experimental groups. Sixteen rats were injected with saline (group 1, n = 9; group 3, n = 7), and 15 rats were injected with 100 mg/kg of agmatine (group 2, n = 8; group 4, n = 7). Injections were given twice: 24 hours and 1 hour before the experiment. Using the modified Langendorf model, rat hearts were perfused with Krebs-Henseleit solution for 105 minutes during phase 1 of the experiment (groups 1 and 2). During phase 2, hearts were exposed to 45 minutes of global ischemia (groups 3 and 4). RESULTS: During phase 1, no statistically significant differences were observed between the agmatine and the control groups. During phase 2, agmatine caused a significant increase in left ventricular pressure (P <.003). At the end of reperfusion, P(max) was 111% +/- 10% from the baseline levels versus only 82% +/- 5% in the control group. After 20 minutes of reperfusion, dP/dt (first-time derivative of the ventricular pressure) in the agmatine group reached full recovery of 106% +/- 12% versus only 64% +/- 14% in the saline group (P =.059). Agmatine also caused a significant increase in coronary flow rate (P <.004) throughout the reperfusion period. Quantitative immunohistochemical staining disclosed reduced cell damage in the agmatine treated hearts (P <.02) versus the control group. CONCLUSION: Agmatine injection given before induced ischemia improves hemodynamic recovery by mechanisms that may be attributed to its vasodilatory properties. PMID- 10684537 TI - Domoic Acid Attenuates the Adenosine-5'-Triphosphate-Induced Increase in AB - BACKGROUND: Although domoic acid (DA), a shellfish neurotoxin, carries a negative surface charge at physiological pH like that of adenosine-5'-triphosphate (ATP), very little is known about its cellular effects. In view of the potentially significant role of extracellular ATP as a signaling molecule for increasing the intracellular concentration of Ca(2+) ([Ca(2+)](i)), we examined the possibility that DA may interfere with this signal transduction mechanism in the myocardium. METHODS AND RESULTS: Cardiomyocytes were isolated from rat heart and loaded with Fura-2 to measure the [Ca(2+)](i). ATP produced a gradual rise in [Ca(2+)](i), reaching a peak level in 25-30 seconds and declining thereafter. DA did not affect the [Ca(2+)](i) in cardiomyocytes; however, it diminished the ATP-induced elevation in [Ca(2+)](i) in the concentration-dependent manner. Kainic acid, an analogue of DA, had a similar effect but at a 25-fold higher concentration, whereas glutamate and aspartate did not modify the action of ATP. Well-known inhibitors of L-type voltage-sensitive Ca(2+) channels, nifedipine and nicardipine, depressed the ATP-induced increase in [Ca(2+)](i), but DA did not produce additive effects with either of these agents. On the other hand, DA potentiated the KCl-induced increase in [Ca(2+)](i) in quiescent cardiomyocytes and augmented the nicardipine-sensitive Ca(2+) transients in electrically stimulated cardiomyocytes. CONCLUSIONS: These results suggest that DA may diminish the ATP-induced increase in [Ca(2+)](i) by inhibiting the ATP interaction with cardiomyocytes in a specific manner. PMID- 10684538 TI - Sustained Intramural Retention and Regional Redistribution Following Local Vascular Delivery of Polylactic-Coglycolic Acid and Liposomal Nanoparticulate Formulations Containing Probucol. AB - BACKGROUND: Probucol reduces restenosis after angioplasty, provided oral administration is begun 1 month before the procedure. Local vascular delivery of a nonoparticulate formulation of probucol may obviate the need for drug loading by acutely raising arterial intramural concentration while providing sustained intramural retention. To test this hypothesis, we compared the retention and redistribution of (35)S-probucol encapsulated in either liposomal or polylactic coglycolic acid (PLGA) nanoparticles after local vascular delivery. METHODS: Nanoparticles were delivered using a Crescendo microporous infusion catheter (Cordis, Warren, NJ) after balloon angioplasty of rabbit iliac arteries (n = 12 18 arteries per formulation per time point). Animals were euthanized on day 0, 3, or 7 after delivery. Iliac arteries, perivascular fat, and downstream tissues were harvested and the radioactivity disintegrations per minute was measured. Autoradiographic and confocal microscopic analyses of tissue sections were performed to evaluate intramural distribution of probucol. RESULTS: Immediately after delivery, radioactivity in the iliac arteries (log[dpm/mg], mean +/- SEM) was greater with PLGA (2.72 +/- 0.08) than with liposomal encapsulation (2.10 +/- 0.08, P = 0.001). Intramural retention of probucol was 23% at 7 days using liposomes and 10% using PLGA, corresponding to a probucol concentration of 0.1 ng/mg tissue for both formulations. By the third day after delivery, radioactivity in peri-iliac fat, femoral arteries, and hindlimb muscle increased by 88%, 29%, and 154%, respectively. Thereafter, radioactivity decreased to 56%, 43%, and 134% of initial dpm respectively, by day 7. CONCLUSIONS: although delivery efficiency was superior with PLGA encapsulation, intramural probucol concentrations were similar on day 7 using both formulations. Radial and axial redistribution of probucol was observed, indicating that this technique can be exploited to increase adjacent tissue delivery. PMID- 10684539 TI - Nifedipine and Bay K 8644 Induce an increase of AB - BACKGROUND: The dihydropyridine-induced vasorelaxation is partly dependent on the endothelium, which does not express L-type calcium channels. Because nitric oxide (NO) is one of the most important endothelium-derived vasorelaxing factors, we investigated how the calcium antagonist nifedipine and the calcium agonist Bay K 8644 modulate intracellular calcium and NO formation in porcine endothelial cells. METHODS AND RESULTS: NO formation of porcine aortic endothelial cell cultures and of native endothelium of intact porcine coronary arteries was measured with an electrochemical electrode, and the intracellular concentration of Ca(2+) [Ca(2+)](i) was evaluated using the Fura-2 technique. Nifedipine induced a concentration-dependent [0,01-1 umol/L] increase in [Ca(2+)](i) and NO formation in cultured porcine aortic endothelial cells, and moreover a dose dependent NO formation in native endothelial cells from intact porcine coronary arteires, which was higher than in cultured cells. This effect was inhibited by N nitro-l-arginine, a specific NO synthase inhibitor. Bay K 8644 caused a [Ca(2+)](i) increase and NO release in cultured cells, too, although to a lesser extent. Nifedipine-induced and Bay K 8644-induced [Ca(2+)](i) rise could be blocked by removal of extracellular calcium, indicating that a calcium influx may be involved. CONCLUSIONS: The calcium antagonist nifedipine as well as the calcium agonist Bay K 8644 cause an increase of [Ca(2+)](i) and NO in porcine endothelium. Therefore, these effects seem to be related to the dihydropyridines as a substance class, which may explain the endothelial component in dihydropyridine-induced vasorelaxation. PMID- 10684540 TI - Electrophysiologic and Antiarrhythmic Effects of Propafenone: Focus on Atrial Fibrillation. PMID- 10684541 TI - Intravenous Amiodarone in the Management of Atrial Fibrillation. AB - BACKGROUND: Although approved only for therapy of life-threatening ventricular tachyarrhythmias, intravenous amiodarone is also being used for the treatment of atrial fibrillation (AF), generally in the intensive care unit setting and most often after cardiac surgery. When used for AF, dosing regimens and clinical experience have varied. METHODS AND RESULTS: This article summarizes representative reports in hopes of clarifying the role of intravenous amiodarone for practitioners who prescribe it for the management of AF. The most immediate and most predictable response is reduction of the ventricular rate, which generally is noted after the first 300-400 mg. Restoration of sinus rhythm (cardioversion) may occur, but the precise incidence in a placebo-controlled, blinded study has not been determined. When present, it often takes 24 hours, and a total dose of 1,000 mg or more. Least certain is the efficacy of the drug in preventing the appearance (when used prophylactically) or reappearance of AF. CONCLUSIONS: More data are required with regard to patient characterization, electrical system status, and dosing regimen to better characterize intravenous amiodarone for this role. PMID- 10684542 TI - Clinical Pharmacology of Carvedilol. AB - BACKGROUND: There is now a wealth of data supporting the use of beta-blockers in heart failure and the additional pharmacological properties of carvedilol are thought to play an important role in the therapeutic efficacy of carvedilol in this disease. METHODS AND RESULTS: Carvedilol is licensed for the treatment of essential hypertension, chronic stable angina, and mild to moderate chronic heart failure. This article provides an up-to-date review of the clinical pharmacology of carvedilol, with particular emphasis on its clinical effects in heart failure. CONCLUSION: Carvedilol is a multiple-action neurohormonal antagonist that offers nonselective beta-blockade, alpha-1 blockade, antioxidant, anti-ischemic mortality, and anti-proliferative properties. In addition to reductions in hospitalization and mortality rates, benefits of carvedilol in heart failure include dramatic improvements in left ventricular function and other parameters of cardiac remodeling. PMID- 10684543 TI - Relative Effects of alpha- and gamma-Tocopherol on Low-Density Lipoprotein Oxidation and Superoxide Dismutase and Nitric Oxide Synthase Activity and Protein Expression in Rats. AB - BACKGROUND: Increasing evidence suggests that vitamin E prevents the progression of atherosclerosis by inhibiting platelet aggregation, monocyte adhesion, and improving plaque stability and vasomotor function. Recently, controversy has arisen as to the relative effects of alpha- and gamma-tocopherol in modulating some mediators of atherosclerosis. METHODS AND RESULTS: We examined the effects of alpha- and gamma-tocopherol on constitutive nitric oxide synthase (cNOS) and superoxide dismutase (SOD) activity and protein expression in rats. Sprague Dawley rats were fed regular chow or chow mixed with alpha- or gamma-tocopherol (100 mg/kg/day) for 7 to 10 days. Plasma alpha- and gamma-tocopherol levels, low density lipoprotein (LDL) oxidation, and cNOS and SOD activity and protein expression were measured. Plasma alpha-tocopherol levels were significantly increased (eP <.01 vs control), but gamma-tocopherol levels fell (P <.01 vs control) in rats fed alpha-tocopherol. Plasma gamma-tocopherol levels were increased (P <.01 vs control), and alpha-tocopherol levels did not change in rats fed gamma-tocopherol. Both alpha- and gamma-tocopherol feeding decreased the rate of LDL oxidation induced by phorbol 12-myristate 13-acetate (PMA)-stimulated leukocytes (P <.01 vs control). Both alpha- and gamma-tocopherol increased SOD activity in plasma and arterial tissues as well as Mn SOD and Cu/Zn SOD protein expression in arterial tissues (all P <.01 vs control). gamma-Tocopherol was more potent than alpha-tocopherol in all these effects (P <.05). Both a- and gamma tocopherol increased NO generation and cNOS activity (all P <.05 vs control). However, only gamma-tocopherol increased cNOS protein expression. CONCLUSIONS: These observations indicate that whereas both alpha- and gamma-tocopherol exert important effects on determinants of oxidationand vasomotor function, effects of dietary gamma-tocopherol supplementation in vivo are less pronounced than those of gamma-tocopherol supplementation. PMID- 10684544 TI - Endothelium-Independent Relaxation of Vascular Smooth Muscle by 17beta-Estradiol. AB - BACKGROUND: Estrogens directly dilate arteries, and this acute relaxation of vascular smooth muscle (VSM) may contribute to the cardioprotective effect of this important hormone. However, the mechanism by which estrogens relax VSM is not clear. METHODS AND RESULTS: Based on observations in isolated smooth muscle cells, we hypothesized that 17beta-estradiol (E(2)) causes dilation through receptor-mediated activation of K(+) channels in VSM cells. To test this hypothesis, E(2)-relaxation was studied in arteries from male Sprague-Dawley rats. We observed that the estrogen receptor antagonist, tamoxifen (3 umol) attenuated E(2) relaxation, suggesting that at least a portion of the relaxation depends on activation of E(2) receptors. The nitric oxide synthase inhibitor, Nomega-nitro-larginine (100 umol) did not affect E(2) relaxation in either denuded or endothelium-intact arterial strips. Furthermore, inhibition of guanylyl cyclase with LY83583 (10 umol) had no effect on the relaxation, suggesting that nitric oxide does not contribute to this relaxation. Vascular segments contracted with 90 mmol KCl to disrupt the K(+) gradient had a similar E(2) relaxation does not require K(+)-channel activation. Finally, E(2) pretreatment inhibited contraction of arterial segments depleted of intracellular calcium but in the presence of extracellular calcium. However, E(2) did not affect contraction of strips in calcium-free solution. CONCLUSIONS: These final experiments suggest that E(2) inhibits Ca(2+) influx but not intracellular calcium release. Together, these studies establish that E(2) causes receptor mediated relaxation of peripheral resistance arteries through inhibition of calcium entry independent of nitric oxide production, guanylyl cyclase stimulation, and K(+)-channel activation. PMID- 10684545 TI - Hypoxic Hypoperfusion Fails to Induce Myocardial Hibernation in Anesthetized Swine. AB - BACKGROUND: Congenital origin of the left coronary artery from the pulmonary artery (ALCAPA) results in chronically dysfunctional myocardium with the partial ability to recover after revascularization. We attempted to establish an ALCAPA syndrome in anesthetized pigs for 24 hours and to compare it with stunned and infarcted myocardium. METHODS AND RESULTS: In group 1 (n = 12), a bypass graft was interposed between the pulmonary artery and the left anterior descending coronary artery (LAD). Reduction of flow in the LAD with gradual increases in flow from the pulmonary artery resulted in an incremental reduction of segment shortening (8.9 +/- 5.3% at 24 hours vs 26.6 +/- 10% at baseline, P <.005). In group 3 (n = 5), 2 cycles of 10-minute LAD occlusion resulted in decreased segment shortening with slow recovery (at 24 hours 18.7 +/- 1.3% vs 24.2 +/- 4% at baseline, segment shortening with slow recovery (at 24 hours 18.7 +/- 1.3% vs 24.2 +/- 4% at baseline, P <.05). In group 3 (n = 6), 1-hour LAD occlusion reduced segment shortening at 24 hours to 4.7 +/- 5.2% (P <.005 vs baseline). Histological analysis of the LAD territory revealed severe degeneration, myolysis, and alteration of the chromatin structure in group 1 comparable to ischemic cell death in group 3, whereas control areas and the LAD area in group 2 showed only minor structural alterations. Infarct size/risk area, as measured by tetrazolium staining, was 49.8 +/- 11.2% in group 1, 9.3 +/- 8.1% in group 2 (P <.005), and 60.3 +/- 9% in group 3. CONCLUSION: Hypoxic myocardial hypoperfusion from the pulmonary artery results in myocardial necrosis in anesthetized pigs. These findings are in contrast to the concept of myocardial hibernation in the ALCAPA syndrome because in this model, hypoxic hypoperfusion failed to induce adaptation to preserve myocardial structure. PMID- 10684546 TI - Superoxide Anion Generation, Superoxide Dismutase Activity, and Nitric Oxide Release in Human Internal Mammary Artery and Saphenous Vein Segments. AB - BACKGROUND: Internal mammary artery (IMA) as conduit for a coronary artery bypass graft (CABG) stays patent longer and more often than saphenous vein (SV). However, the precise differences in the biology of IMA and SV are unclear. METHODS AND RESULTS: To examine inherent difference in superoxide anion, superoxide dismutase (SOD) and nitric oxide (NO) formation in IMA and SV as a basis for differences in patency rates, we measured these parameters in vascular segments of patients undergoing CABG. Superoxide anion generation was measured by lucigenin chemiluminescence and reduction of cytochrome c, SOD by inhibition of pyrogallol auto-oxidation, and No as nitrite/nitrate fluorometrically using 2-3 diaminonaphthalene as a probe. Generation of superoxide anion, SOD activity, and No formation were all greater in the IMA than in the SV segments (IMA:SV = 2.6:1, 2.9:1, 1, and 3.0:1, respectively, all P <.010. There was a positive correlation between superoxide anion generation and SOD activity (r = 0.65, P <.05; r = 0.70, P <.05 in IMA and SV, respectively) and NO release (r = 0.68, P <.05; r = 0.75, P <.03 in IMA and SV, respectively). Western blot analysis showed no differences in SOD and NO synthase protein expression in IMA and SV segment homogenates. To examine whether greater superoxide anion generation, SOD activity, and NO formation are unique to IMA, we studied pulmonary artery (PA) and pulmonary vein (PV) segments taken from patients undergoing lung resection. Superoxide anion generation, SOD activity, and NO formation were also found to be greater in PA than in PV segments. CONCLUSIONS: Inherently greater superoxide anion generation and subsequently increased formation of SOD and NO release in IMA (vs SV) may be a factor in the greater patency of the former as CABG conduit. Because both IMA and PA are exposed to pulsatile stretch and cary blood at higher pressure than the SV and PV, it is likely that these 2 factorsd account for the observed differences. PMID- 10684547 TI - Viagra and Cardiovascular Disease. AB - BACKGROUND: The introduction of the drug sildenafil (Viagra; Pfizer, New York, NY) into the armamentarium for treatment of erectile dysfunction is a major advance. Many of the patients who will benefit from its use have cardiovascular disease. Erectile dysfunction and cardiovascular disease share common risk factors. Although the metabolic demands of sexual activity are modest and the associated risk for coronary events is low, the clinician caring for cardiac patients needs to be aware of the pharmacology and hemodynamic profile of sildenafil in those with heart disease who use cardioactive drugs. METHODS AND RESULTS: We reviewed the current literature relating to the pharmacology, hemodynamic profile, efficacy, safety, and clinical application of sildenafil in patients with cardiovascular disease. Sildenafil is highly effective in the treatment of erectile dysfunction. The overall incidence of cardiovascular adverse events is low and similar to placebo. Current postmarketing data do not suggest an increase in cardiovascular death in sildenafil users. The drug is contraindicated in those taking organic nitrates. It should be used with caution and on an individual basis in patients who have active coronary ischemia and heart failure with tenous blood pressure and volume status. CONCLUSIONS: When used with discretion, sildenafil is safe, effective, and has the potential to greatly enhance quality of life in the relatively large proportion of the population with heart disease. PMID- 10684548 TI - Primary Hyperparathyroidism and Polymorphic Ventricular Tachycardia. AB - BACKGROUND: Primary hyperparathyroidism is a rare but important cause of ventricular arrhythmias. METHODS AND RESULTS: The medical records of a patient with primary hyperparathyroidism and polymorphic ventricular tachycardia was reviewed. The patient was serially interviewed and examined before and after parathyroidectomy. A literature search was performed. The association of primary hyperparathyroidism and polymorphic ventricular tachycardia is rare. Hypokalemia and hypomagnesemia occur infrequently in primary hyperparathyroidism. CONCLUSIONS: The potential for malignant ventricular arrhythmias in the setting of primary hyperparathyroidism must be recognized. PMID- 10684549 TI - Preface PMID- 10684550 TI - Endoscopic treatments for Zenker's diverticulum. PMID- 10684551 TI - Cardiomyotomy. AB - During the last decade, minimally invasive surgery has replaced open surgery in the treatment of esophageal achalasia. This new approach, in fact, determines results similar to the open approach, but is associated to a shorter hospital stay, minimal postoperative discomfort, and faster return to regular activity. Between 1991 and 1998, 168 patients underwent a cardiomyotomy by minimally invasive techniques. Good or excellent results were obtained in 85% of patients after thoracoscopic myotomy, and 93% of patients after laparoscopic myotomy and partial fundoplication. The latter procedure was followed by a lower incidence of postoperative gastroesophageal reflux (60% versus 17%). Laparoscopic Heller myotomy and partial fundoplication has emerged as the procedure of choice for esophageal achalasia, and it should be considered today the primary form of treatment for this disease. PMID- 10684552 TI - Current state, techniques, and results of laparoscopic antireflux surgery. AB - The introduction of laparoscopic fundoplication has dramatically changed the face of antireflux surgery. Central to the success of laparoscopic fundoplication is careful preoperative patient evaluation and attention to surgical technique. Emerging evidence has questioned the long-term durability of laparoscopic partial fundoplications underscoring the place of laparoscopic Nissen fundoplication as the procedure of choice for most patients. The technique of laparoscopic Nissen fundoplication should incorporate crural closure, complete fundic mobilization by short gastric vessel division, and the creation of a short, loose fundoplication by enveloping the anterior and posterior fundic walls around the esophagus. Relief of typical reflux symptoms can be anticipated in over 90% of patients. The outcome of atypical reflux symptoms is less predictable, on average two thirds of patients benefiting. The cost of laparoscopic fundoplication compares favorably to long-term medical therapy and open fundoplication. Current trends indicate that laparoscopic fundoplication is being used increasingly as an alternative to long-term medical therapy. PMID- 10684553 TI - Laparoscopic treatment of large hiatal hernias. AB - Large hiatal or paraesophageal hernias constitute between 5% and 10% of all hiatal hernias. This hernia is a potential threatening complication, and a timely operative correction should be performed in all patients with an acceptable risk. Based on the lessons learned from conventional approach, laparoscopic treatment has confirmed the initial good results with all advantages of laparoscopic surgery. Reduction of the hernia, excision of the sac, and approximation of the hiatus followed by selective use of an antireflux procedure and some form of gastropexy constitute the operative steps to obtain optimal postoperative results. PMID- 10684554 TI - Laparoscopic gastric drainage procedures. AB - Gastric outlet obstruction continues to be an indication for drainage despite the common use of powerful proton pump inhibitors. Minimal invasive surgery techniques now play a significant role in the treatment of this pathology. Complicated peptic ulcer disease and cancer are the two most common causes. To accomplish drainage, advanced laparoscopic techniques are required. A variety of procedures are possible, and these are discussed in detail in this report. The advantages of the laparoscopic approach have been realized in this group of patients. PMID- 10684555 TI - Ultrasonic dissectors and minimally invasive surgery. AB - As increasingly complex operations are performed laparoscopically, new problems arise regarding basic tasks such as dissection and retraction. Emerging technologies continue to reduce the technical demands of minimally invasive surgery. Recent studies have shown that ultrasonic devices have the potential to replace electrocautery without compromising safety in minimally invasive operations. With the combination of several functions into a single instrument, significant reductions in operative time and expense are possible and should increase the acceptance of this new technology. PMID- 10684556 TI - Immunocytochemical localization of actin and tubulin in the integument of land crab (Gecarcinus lateralis) and lobster (Homarus americanus). AB - The crustacean integument consists of the exoskeleton and underlying epithelium and associated tissues. The epithelium, which is composed of a single layer of cells, is responsible for the cyclical breakdown and synthesis of the exoskeleton associated with molting (ecdysis). During premolt (proecdysis) the epithelial cells lengthen and secrete the two outermost layers (epicuticle and exocuticle) of the new exoskeleton while partially degrading the two innermost layers (endocuticle and membranous layer) of the overlying old exoskeleton. This increased cellular activity is associated with increased protein synthesis and a change in cell shape from cuboidal to columnar. The cytoskeleton, composed of microfilaments (actin) and microtubules (tubulin), plays important roles in the intracellular organization and motility of eukaryotic cells. Immunoblot analysis shows that the land crab exoskeleton contains actin, tubulin, and actin-related proteins (Varadaraj et al. 1996. Gene 171:177-184). In the present study, immunocytochemistry of land crab and lobster integument showed that both proteins were localized in various cell types, including epithelia, connective tissue, tendinal cells, and blood vessels. Muscle immunostained for actin and myosin, but not for tubulin. The membranous layer of land crab (the other layers of the exoskeleton were not examined) and membranous layer and endocuticle of lobster also reacted specifically with anti-beta-actin and anti-alpha-tubulin monoclonal antibodies, but not with an anti-myosin heavy chain antibody. During proecdysis immunolabeling of the membranous layer decreased probably due to protein degradation. The staining intensity for actin and tubulin in the proecdysial epithelium was similar to that in the intermolt (anecdysial) epithelium, suggesting that there was a net accumulation of both proteins proportional to the increase in cellular volume. These results support the previous biochemical analyses and, more specifically, localize actin and tubulin in exoskeletal structures, suggesting that they may serve both intracellular and extracellular functions in crustaceans. J. Exp. Zool. 286:329-342, 2000. PMID- 10684557 TI - Intra- and extracellular dehydration has no effect on plasma levels of angiotensin II in an amphibian. AB - Previous studies have demonstrated that both dehydration (intra and extracellular) and treatment with angiotensin II (A-II) induce changes in thirst related behavior in the spadefoot toad, Scaphiopus couchii. One of the steps in determining a causal relationship between a hormone and a behavior is to determine that there is association between an animal's performance of the behavior and changes in endogenous hormonal concentrations. The hypothesis tested that plasma levels of the peptide hormone A-II would change as a result of dehydration known to induce water absorption response (WR) behavior in the spadefoot toad. Plasma samples were taken from toads dehydrated intracellularly by injection of hypertonic solutions of NaCl or sucrose at levels known to induce WR behavior. As an osmotic control, a group of animals was injected with urea, which has been demonstrated to not induce WR behavior. In order to determine the effects of extracellular dehydration on plasma, A-II levels in toads dehydrated by plasma volume depletion via cardiac puncture were compared to sham-punctured controls. None of the treatments in any experiment resulted in significant differences in plasma levels of angiotensin II among groups sampled at the time when WR behavior occurs. These results do not support the hypothesis that dehydration-induced thirst is stimulated by changes in plasma A-II concentrations at the onset of WR behavior. J. Exp. Zool. 286:343-349, 2000. PMID- 10684558 TI - Anatomy and physiology of neurons composing the commissural ring nerve of the cricket, Acheta domesticus. AB - The commissural ring nerve (RN) of the cricket Acheta domesticus links together the two cercal motor nerves of the terminal abdominal ganglion. It contains the axons of almost 100 neurons including two bilateral clusters of eight to 13 ventrolateral neurons and approximately 75 neurons with midline somata within the terminal abdominal ganglion. The ventrolateral neurons have an ipsilateral dendritic arborization within the dorsal neuropil of the ganglion and their axons use the RN as a commissure in order to enter the contralateral nerves of the tenth ganglionic neuromere. In contrast, most midline neurons have bifurcating axons projecting bilaterally into the neuropil of the ganglion as well as into the RN where they often branch extensively before entering the contralateral tenth nerves. Most RN neurons have small, non-spiking somata with spike initiation zones distant from the soma. Many midline neurons also produce double peaked spikes in their somata, indicative of multiple spike initiation zones. Spontaneous neuronal activity recorded extracellularly from the RN reveals several units, some with variable firing patterns, but none responding to sensory stimuli. The RN is primarily composed of small (50 nm diameter) axon profiles with a few large (0.5-1 microm diameter) profiles. Occasionally, profiles of nerve terminals containing primarily small clear vesicles and a few large dense vesicles are observed. These vesicles can sometimes be clustered about an active zone. We conclude that the primary function of the RN is to serve as a peripheral nerve commissure and that its role as a neurohemal organ is negligible. J. Exp. Zool. 286:350-366, 2000. PMID- 10684559 TI - Triggering of cryoprotectant synthesis in the woolly bear caterpillar (Pyrrharctia isabella Lepidoptera: Arctiidae). AB - Isabella tiger moths (Pyrrharctia isabella) overwinter as caterpillars (i.e., woolly bears) that can survive freezing at moderate subzero temperatures. We observed an increase in hemolymph osmolality for field-collected woolly bears during October (325 +/- 47 to 445 +/- 27 mOsmol/liter) and tested the influence of temperature and moisture levels on cryoprotectant production. Laboratory acclimation was done at 5 degrees C in moist conditions and at 25 degrees C acclimation in both dry and moist conditions. Body water contents were diminished by dehydration at 25 degrees C for 4 days (57 +/- 4%). Caterpillars collected in early October did not alter their hemolymph osmolality during cold acclimation, but caterpillars increased by 45% (to 647 +/- 90 mOsmol/liter) after 4 days at 5 degrees C following their collection in late October. Hemolymph composition was markedly changed in caterpillars experiencing dehydration at 25 degrees C (1042 +/- 200 mOsmol/liter; 507 +/- 225 mmol glycerol/liter), whereas caterpillars showed no change in their hemolymph composition when kept moist at 25 degrees C. Our experiments reveal that both dehydration and cold acclimation rapidly induce cryoprotectant synthesis in P. isabella caterpillars. J. Exp. Zool. 286:367-371, 2000. PMID- 10684560 TI - Food chemical discriminations by an herbivorous lizard, Corucia zebrata. AB - Adjustment of chemosensory response to diet should be apparent in evolutionary changes corresponding to dietary shifts. Because most lizards are generalist predators of small animals, relationships between chemosensory behavior and diet are difficult to detect. Nevertheless, the evolution of herbivory by a small number of lizards provides an opportunity to detect any corresponding change in response to plant chemicals. I studied tongue-flicking and biting by the large, herbivorous scincid lizard Corucia zebrata in response to chemical cues from crickets, romaine lettuce, and control stimuli presented on cotton swabs. The skinks exhibited significantly stronger response to plant and animal chemicals than to controls for several variables: greater number of individuals that bit swabs, shorter latency to bite, greater rate of tongue-flicks, and greater tongue flick attack score. The clearest differences were observed for tongue-flick attack score, a composite variable that combines the effects of tongue-flicking and biting. An insectivorous member of the same subfamily, Scincella lateralis, shows strong tongue-flicking and biting response to chemical prey cues, but not to plant chemicals. This suggests that response to plant chemicals by C. zebrata may have evolved in tandem with the incorporation of plants into the diet and that response to cricket chemicals has been retained, perhaps due to similarities between plant and animal food. The findings support the hypothesis that dietary shifts induce corresponding changes in chemosensory response, but provide only a single independent contrast for a study of correlated evolution between plant diet and chemosensory response to plants. J. Exp. Zool. 286:372-378, 2000. PMID- 10684561 TI - Embryonic muscle development in rainbow trout (Oncorhynchus mykiss): a scanning electron microscopy and immunohistological study. AB - Embryonic muscle development was studied in rainbow trout (Oncorhynchus mykiss) at low and high temperature using scanning electron microscopy (SEM) and immunohistology. Somite development was described starting at stage 16 (Vernier JM. 1969. Ann Embryol Morphogen 4:495-520) for both temperatures, with special interest in their shape and size. Muscle differentiation, associated with somite growth, is characterized by a larger increase in height compared to width and by acquisition of a chevron shape. Thin structures such as striation, sarcomeres, and myofibrils within muscle cells and myotubes were observed starting at the eyed stage (stage 24). Immunohistological analyses showed appearance of embryonic fast myosin at stage 20 in the deep part of the somite. The area where myosin was expressed extended in the somite throughout embryonic development and the presence of myosin was observed in the entire somite at hatching (stage 30). Slow myosin was expressed in a monolayer of superficial cells at the eyed stage and during the entire embryonic development. Then it was expressed in a few layers of cells located in the red muscle area. These results suggest that muscle differentiation, characterized by myosin expression, is engaged at stage 20. Myogenesis starts in the deep part of the somite, near the notochord and progresses laterally to cover the complete somite at hatching when the somite is composed of muscle fibres exhibiting a high degree of maturity. No significant difference was observed in terms of muscular development between low- and high temperature conditions. J. Exp. Zool. 286:379-389, 2000. PMID- 10684562 TI - Contrasting stress response of male arctic ground squirrels and red squirrels. AB - A hormonal-challenge protocol was used to compare the stress response of males of Arctic ground squirrels and red squirrels during the breeding season (May). These squirrels live in the same boreal forest of the Yukon, but have very different life histories and utilize the forest in markedly different ways. Red squirrels had levels of total cortisol, maximum corticosteroid-binding capacity, and free cortisol that were 5, 7, and 2 times, respectively, those of Arctic ground squirrels. Red squirrels were resistant to suppression by an artificial glucocorticoid, dexamethasone (DEX); Arctic ground squirrels were not. Cortisol levels in red squirrels responded slowly but continuously to the ACTH injection; Arctic ground squirrels responded rapidly and then stabilized. Testosterone levels in red squirrels were extremely sensitive to the challenge, being suppressed by both DEX and ACTH; levels in Arctic ground squirrels were resistant to the challenge, being modestly suppressed by DEX and stimulated by ACTH. Energy mobilization, as measured by glucose and free fatty acid responses, was not affected. Red squirrels had four times the levels of white blood cells and higher proportions of lymphocytes and lower proportions of eosinophils than Arctic ground squirrels, indicating that the latter were in worse condition immunologically. Our evidence suggests that the functions associated with the hypothalamic-pituitary-adrenal axis are compromised in breeding male Arctic ground squirrels, but not in red squirrels. We propose that in male red squirrels this axis has evolved in the context of a stable social system based on long lived animals with individual territories which are needed to deal with unpredictable winter food supplies. In contrast, Arctic ground squirrels escape the rigors of winter by hibernation and this hormonal axis has evolved in short lived males in the context of intense intra-sexual competition in a social system based on female kin groups and regular male dispersal to avoid inbreeding. J. Exp. Zool. 286:390-404, 2000. PMID- 10684563 TI - Changes along the pituitary-gonadal axis during maturation of the black carp, Mylopharyngodon piceus. AB - The black carp, Mylopharyngodon piceus, is a late-maturing cyprinid reaching sexual maturity at the age of 6-7 years. The present work attempted to define nonfunctional sites along the pituitary-gonadal axis in immature fish utilizing in vivo and in vitro challenge experiments. Two- and 3-year old fish injected with salmon gonadotropin-releasing hormone analog (sGnRHa; 10 microg/kg) and metoclopramide (20 mg/kg) did not reveal any increase in circulating gonadotropin (cGtH) or estradiol (E(2)) level. Furthermore, cGtH release from cultured pituitary cells of fish at these ages did not increase in response to sGnRH (0.1 nM - 1 microM) but was augmented when exposed to TPA (12.5 nM). However, 4-year old female fish did respond to the above treatments both in vivo and in vitro. These results suggest the existence of nonfunctional site(s) proximal to the activation of PKC in the immature black carp gonadotrophs, probably at the level of GnRH receptors. These site(s) start to become functional in 4-year old fish. Two- and 3-year old fish injected with common carp pituitary extract (CPE) containing 350 microg cGtH/kg did not show any increase in circulating E(2). In addition, the estrogen secretion from fragments of the rudimentary gonads did not increase after exposure to CPE containing cGtH (0.5-4 microg/ml) but was elevated dose-dependently by exposure to dbcAMP (0.3-3 mM). However, the ovaries of 4-year old fish did respond to the gonadotropic stimulation, both in vivo and in vitro. These results suggest the existence of other non-functional site(s) in the immature black carp, proximal to the formation of cAMP in the gonads, probably at the level of GtH receptors. These site(s) start to become functional in 4-year old females. Another source of E(2) was discovered in the immature black carp: namely, the fat pad adjacent to the gonads. In contrast to the visceral adipose tissue, the fat pad secretes estrogen in response to cAMP elevation in 2- and 3 year old fish while in 4-year old fish it also responds to gonadotropic stimulation. Due to its large mass and high steroidogenic potency, it is assumed that the gonadal fat pad is involved in the process of puberty in the black carp. J. Exp. Zool. 286:405-413, 2000. PMID- 10684564 TI - Cilia-driven rotational behavior in gastropod (Physa elliptica) embryos induced by serotonin and putative serotonin reuptake inhibitors (SSRIs). AB - We characterized the serotonin (5-hydroxytryptamine; 5-HT) receptor mediating cilia-driven rotational movement in embryos of the freshwater gastropod Physa elliptica. In addition, putative serotonin reuptake inhibitors (SSRIs), previously shown to induce other 5-HT-mediated processes in molluscs, were tested for their ability to induce rotation. As in previous studies with other freshwater gastropods, 5-HT induced a significant dose-dependent increase in rotation from 10(-6) to 10(-4) M. The 5-HT(1A) agonist 8-OH-DPAT produced a similar dose-dependent increase in rotation. However, the 5-HT(2) agonist alpha CH3-serotonin evoked a significant rotational response only at the highest concentration of 10(-4) M. The 5-HT(2) receptor antagonist mianserin not only blocked 5-HT-induced rotation, it reduced rotation rate below that of baseline. However, two other antagonists, cyproheptadine (5-HT(2)) and propranolol (5 HT(1)), caused similar responses that consisted of an initial rotational surge followed by reduced rotation. Thus, these drugs appear to act as partial agonists. The putative SSRI fluvoxamine exhibited a significant dose-dependent increase in positive rotation as that seen with 5-HT. The SSRIs paroxetine and fluoxetine both caused an increase in rotation at 10(-6) and 10(-5) M but reduced rotation rate below that of baseline at 10(-4) M. These results agree with other studies on aquatic molluscs, suggest a 5-HT receptor with a mixed 5-HT(1)/5-HT(2) pharmacological profile and add to a now growing body of literature on the pharmacology of molluscan 5-HT receptors. In addition, all the tested putative SSRIs induced cilia-driven rotation in Physa embryos, indicating either the presence of 5-HT reuptake transporters or that these compounds act as 5-HT receptor ligands. J. Exp. Zool. 286:414-421, 2000. PMID- 10684565 TI - Influence of incubation temperature on morphology, locomotor performance, and early growth of hatchling wall lizards (Podarcis muralis). AB - Eggs of wall lizards (Podarcis muralis) were incubated at three temperatures approaching the upper limit of viability for embryonic development in this species (26, 29, and 32 degrees C) to assess the influence of temperature on various aspects of hatchling phenotype likely affecting fitness. The thermal environment affected size and several morphometric characteristics of hatchling lizards. Hatchlings from eggs incubated at 32 degrees C were smaller (snout-vent length, SVL) than those from 26 and 29 degrees C and had smaller mass residuals (from the regression on SVL) as well as shorter tail, head, and femur relative to SVL. Variation in the level of fluctuating asymmetry in meristic and morphometric traits associated with incubation temperatures was quite high but not clearly consistent with the prediction that environmental stress associated with the highest incubation temperatures might produce the highest level of asymmetry. When tested for locomotor capacity in trials developed at body temperatures of 32 and 35 degrees C, hatchlings from the 32 degrees C incubation treatment exhibited the worst performance in any aspect considered (burst speed, maximal length, and number of stops in the complete run). Repeated measures ANCOVAs (with initial egg mass as covariate) of snout-vent length and mass of lizards at days 0 and 20 revealed significant effects of incubation temperature only for mass, being again the hatchlings from eggs incubated at 32 degrees C those exhibiting the smallest final size. All together, our results evidenced a pervasive effect of thermal regime during incubation (and hence of nest site selection) on hatchling phenotypes. However, incubation temperature does not affect hatchling phenotypes in a continuous way; for most of the analysed traits a critical threshold seems to exist between 29 and 32 degrees C, so that hatchlings incubated at 32 degrees C exhibited major detrimental effects. J. Exp. Zool. 286:422-433, 2000. PMID- 10684566 TI - Novel aspects of the activities and subcellular distribution of enzymes of ketone body metabolism in the liver and kidney of the goldfish, Carassius auratus. AB - The metabolic organization of ketone body metabolism of liver and kidney of the goldfish Carassius auratus was assessed by measuring maximal activities, subcellular distribution, and stereoisomer preference of ketone body enzymes. These determinations indicate that the organization of ketone body metabolism in liver and kidney of goldfish differs from that of mammals in some respects. All the enzymes of ketone body metabolism were present in liver and kidney of goldfish, with the exception of hydroxymethylglutaryl-CoA (HMG-CoA) synthetase, which was not detected in liver. Two forms of beta-hydroxybutyrate dehydrogenase (betaHBDH) with different stereospecificity for beta-hydroxybutyrate (D- and L beta-hydroxybutyrate) were detectable in liver and kidney. All of the ketone body enzymes measured in liver were mainly in the mitochondrial fraction, with the exception of D- and L-betaHBDH, which were cytosolic. In kidney, HMG-CoA synthase, together with HMG-CoA lyase and acetoacetyl CoA thiolase (AcoAT), were found mainly in the mitochondrial fraction. L-betaHBDH was mainly cytosolic in kidney, but by contrast with liver, D-betaHBDH was mainly found in the mitochondria, and SKT was distributed in both the mitochondrial and cytosolic compartments. J. Exp. Zool. 286:434-439, 2000. PMID- 10684567 TI - Disrupting the geranylgeranylation at the C-termini of the shrimp Ras by depriving guanine nucleotide binding at the N-terminal. AB - In order to assess the effects of guanine nucleotide binding on the geranylgeranylation at the CAAX box of the shrimp Ras, we experimented with the shrimp Penaeus japonicus Ras (S-Ras) which is geranylgeranylated at the C termini, shares 85% homology with mammalian K(B)-Ras protein and demonstrates identity in the guanine nucleotide binding domains (Huang C-F, Chuang N-N. 1999. J Exp Zool 283:510-521). Several point mutations in the S-ras gene were generated at codons 12 (G12V), 61 (Q61K), and 116 (N116I). The bacterially expressed mutant S-Ras proteins, G12V and Q61K, were bound with GTP without hydrolysis. In contrast, the mutant S-Ras N116I was defective in its ability to bind any guanine nucleotides. Autoradiography studies showed that the purified shrimp protein geranylgeranyltransferase I (Lin R-S, Chuang N-N. 1998. J Exp Zool 281:565-573) was unable to catalyze the transfer of [(3)H]-geranylgeranylpyrophosphate to this mutant N116I but very competently caused the geranylgeranylation of GTP-locked mutants, G12V and Q61K. These results demonstrate that the geranylgeranylation at the CAAX box of the shrimp Ras protein requires the proper binding of guanine nucleotide at its N-terminal region. J. Exp. Zool. 286:441-449, 2000. PMID- 10684568 TI - Lack of physiological plasticity in the early chicken embryo exposed to acute hypoxia. AB - By exposing chicken embryos to hypoxia (10%) acutely (2, 4, and 6 hr) during early development (2, 3, and 4 days) we tested the hypothesis that hypoxia has an impact on embryonic growth and impairs cardiac development at the time cardiac morphogenesis is taking place. After the hypoxic perturbation, the embryos were allowed to develop until day 9, when embryo mass, heart mass, and rate of oxygen consumption were recorded. Four-day-old embryos exposed to 6 hr of hypoxia showed an increased mortality (38.9% versus 18% for controls), indicating the immediate effect of hypoxia on survivability. While only 8% of the controls displayed morphological abnormalities, 3- and 4-day-old embryos exposed for 6 hr showed more frequent developmental abnormalities (25% and 30% respectively). No significant differences in embryo or heart mass were found except in 4-day-old embryos exposed for 2 hr. Mass-specific oxygen consumption was not different between controls and embryos exposed to hypoxia at 2 or 3 days of development, but it was increased in 4-day-old embryos exposed for 4 hr (P < 0.05). These results suggest that an acute hypoxic episode does not have an impact when occurring very early in development (days 2 or 3). However, when the hypoxic episode occurs on day 4, survivability is largely decreased. Considering the lack of permanent effects on the surviving embryos, we suggest that the early embryo resorts to a simple strategy of death or survival, and the individual capacity for survival must be based on interindividual differences rather than the existence of compensatory mechanisms. J. Exp. Zool. 286:450-456, 2000. PMID- 10684569 TI - Participation of the GM1 ganglioside in the gastrulation of anuran amphibian Bufo arenarum. AB - In the present paper we established the ganglioside composition of the blastula and gastrula stages of the anuran amphibian Bufo arenarum, two relevant stages characterized by dynamic changes in morphology and cellular rearrangements. Densitometric studies evidenced that GD1a and GT1b were the more abundant gangliosides of the blastula embryos whereas GM1 and GM2 were the predominant species in gastrula embryos. Analysis of ganglioside abundance indicates that the "a" and "b" synthesis pathways perform similar biosynthetic activities in the blastula stage, in contrast to the gastrula stage in which a marked predominance of the "a" pathway occurred. The spatio-temporal expression of GM1 and of polygangliotetraosyl ceramides (pGTC) was investigated by wholemount immunocytochemistry using cholera toxin B subunit (CTB) and an affinity purified human anti-GM1 antibody. The pGTC were detected as GM1 after treatment with neuraminidase. Blastomeres from the inner surface of the blastocoelic roof (BCR) of blastula embryos were GM1 and pGTC positive. At midgastrula stage, embryos showed an increased labeling on the inner surface of BCR. To establish whether the GM1 ganglioside was involved in the gastrulation processes, CTB, anti-GM1 antibodies and anti-GM1 Fab' fragments were microinjected into the blastocoel cavity of blastula embryos. Treatment with the probes blocked gastrulation. Scanning electron microscopy analysis of blocked embryos revealed that mesodermal cell migration, radial interdigitation, and convergent extension movements were affected. The blocking of gastrulation was correlated with the absence of fibronectin and EP3/EP4 on the inner surface of blastocoelic roof of CTB- or anti GM1 treated embryos. Results show that the GM1 ganglioside is differentially expressed by embryonic cells and participates in the morphogenetic processes of amphibian gastrulation. J. Exp. Zool. 286:457-472, 2000. PMID- 10684570 TI - Corticotropin-releasing factor accelerates metamorphosis in Bufo arenarum: effect on pituitary ACTH and TSH cells. AB - The actions of several neuropeptides as hypothalamic mediators in the regulation of Bufo arenarum metamorphosis were investigated. Prometamorphic larvae were injected with 1.5 microg thyrotropin-releasing hormone (TRH), 2 microg ovine corticotropin-releasing factor (oCRF), 2 microg mammalian gonadotropin-releasing hormone (mGnRH), 2 microg human growth hormone-releasing hormone (hGHRH), or Holtfreter solution (control group). Larvae received two injections with the same dose: one at the beginning of the experiment and the other 7 days later. Several morphologic parameters (total length, tail length, wet weight, hind limb length, and metamorphic stages) were measured as indicators of growth and metamorphic development. These measurements were taken in 20 larvae per treatment or control group at the beginning of the experiment, at day 7 and at day 14 when the experiment ended. We observed that only the administration of exogenous CRF stimulated resorption of the tail and accelerated the rate of metamorphosis. In the pituitary of CRF-treated larvae we observed that thyrotropin (TSH) and adrenocorticotropic hormone (ACTH) producing cells showed a weaker immunoreactivity, a decrease in cell number and a reduction of volume density when compared with normal larvae. In conclusion, the results obtained indicate a possible role for CRF in Bufo arenarum metamorphosis. CRF may regulate interrenal and thyroid activity by acting directly upon TSH and ACTH cells. On the other hand, TRH, GnRH and GHRH were inactive in stimulating growth or metamorphosis of Bufo arenarum. J. Exp. Zool. 286:473-480, 2000. PMID- 10684571 TI - Regulation of opioid peptides on the release of arginine vasotocin in the hen. AB - Arginine vasotocin (AVT), an avian neurohypophysial hormone, is released during osmotic stimulation and oviposition. In the present study, the role of opioid peptides on AVT release was studied by examining the effects of an opioid agonist and antagonist on osmotic- and oviposition-induced secretion of AVT. The administration of hypertonic saline (1.5 M NaCl) induced an increase in the plasma levels of AVT. The simultaneous administration of morphine, an opioid receptor agonist, inhibited the osmotically induced increase in plasma levels of AVT in a dose-dependent manner. On the other hand, the co-administration of morphine with naloxone, an opioid receptor antagonist, attenuated the inhibitory effect of morphine. Moreover, injection of naloxone alone enhanced the osmotically induced increase in plasma levels of AVT. However, the administration of morphine did not inhibit the oviposition-induced increase in plasma levels of AVT. These results suggest that osmotic-induced release of AVT may be under opioid regulation, while oviposition-induced release of AVT may be controlled by a different mechanism. J. Exp. Zool. 286:481-486, 2000. PMID- 10684572 TI - Interrelationship between food availability, fat body, and ovarian cycles in the frog, Rana tigrina, with a discussion on the role of fat body in anuran reproduction. AB - Long-term experiments were conducted to study the progression of vitellogenic cycles in Rana tigrina (an annual breeder) having different foraging backgrounds and held under conditions of weekly or daily food supply and in presence or absence of abdominal fat bodies. They were autopsied in June to assess fecundity. In nature an adult R. tigrina produces on an average 4,000 eggs/100 g body mass (b.m.) And spawns in June-July following monsoon rains. Weekly feeding from July to next breeding season, June resulted in a significant decrease in both fecundity (1700 eggs/100 g body b.m.) And mean size of eggs, compared to well-fed or wild-caught frogs. The abdominal fat bodies were barely seen in frogs fed weekly throughout, whereas in frogs fed weekly from July-December but daily from January onwards, the fat bodies became noticeable (1% of b.m.) And number and mean size of eggs increased significantly over those fed weekly throughout. Frogs captured in January possessed enlarged fat bodies (5% of b.m.), depicting a good foraging history. Maintenance of these frogs on a weekly feeding regimen led to an exhaustion of fat stores. They produced less number of eggs (2, 000/100 g b.m.) As compared to wild frogs but of normal size, whereas daily feeding slowed down a depletion of fat body mass and also significantly increased fecundity (3,000/100 g b.m.) Over the weekly fed individuals. Sham operation or fat body ablation in October or February had no significant effect on total fecundity per se (3,000-3,500 eggs/100 g b.m.) Compared to that of wild-caught frogs. However, eggs were significantly smaller due to fat body ablation despite daily feeding. The study shows that food abundance/fat bodies influence egg size and number in R. tigrina and that a direct or indirect functional relationship exists between fat body and ovarian cycles that are characteristically inverse to each other. J. Exp. Zool. 286:487-493, 2000. PMID- 10684573 TI - Visual and nutritional food cues fine-tune timing of reproduction in a neotropical rainforest bird. AB - Food may act as a proximate factor in the regulation of avian seasonal breeding. Food cues could provide particularly important seasonal information to birds living in variable tropical environments, but this has not yet been tested. Spotted antbirds (Hylophylax n. naevioides) inhabiting a humid forest in central Panama (9 degrees N) likely use changes in the tropical photoperiod to time reproduction on a long-term, seasonal basis. We predicted that these insectivorous birds also adjust reproduction to short-term cues such as food availability because the onset of the rainy season and the resulting increase in insect abundance varies considerably between years. To test this prediction, prior to their breeding season (when they had half-maximal gonads), we either exposed captive male spotted antbirds to an ad libitum standard diet only or added live crickets to this diet. Males that received live crickets significantly increased gonad sizes within 3 weeks over controls on the standard diet. Moreover, in six additional experiments cricket availability always increased song rate, usually within a few days. The stimulatory effect of live crickets on song activity may function independent of nutritional aspects: Freshly killed crickets, providing similar nutritional content as live crickets, did not stimulate the birds' song activity. However, song activity increased to intermediate levels when live crickets were shown under a clear plastic wrap, i.e., when birds could see but not eat crickets. We hypothesize that the opportunity to see and handle live insects stimulates song and reproductive activity in these birds. Our data indicate for the first time that a tropical rainforest bird can use food cues to evaluate the suitability of local environmental conditions for breeding. J. Exp. Zool. 286:494-504, 2000. PMID- 10684574 TI - Gonadal morphology of female diploid gynogenetic and triploid rainbow trout. AB - Chromosome sets of fishes can be manipulated; this practice includes the production of triploid and gynogenetic salmonids. Such chromosomal modifications often result in abnormal ovarian development. In rainbow trout (RBT), triploid females have string-like gonads lacking significant developing oocytes and are suggested to be sterile due to the odd set of chromosomes disrupting oogenesis. Aberrant ovarian development is reported to occur in about 30% of gynogenetic females. It has been suggested that gynogenetic fish are more prone to expressing developmental abnormalities due to either increased homozygosity or to incomplete inactivation of the paternal chromatin. This investigation was done to compare the ovarian morphology of female triploid and induced gynogenetic diploid RBT. The objective was to determine whether the presence of supernumerary chromosomal fragments, potentially generated during the process of sperm genome inactivation, would result in abnormal gonadal development in gynogens comparable to that observed in triploid females. Gonadal morphology was observed and karyotypical analysis was completed on 21 gynogenetic fish. In 90% of the fish examined, the presence of chromosomal fragments was positively correlated with irregular ovarian development. The atypical gonadal morphology observed in the gynogens resembled triploid RBT ovarian morphology. The results of this investigation support the hypothesis that disruption of the normal diploid chromosomal complement alters germ cell development in gynogenetic female RBT due to the unbalanced nature of the genome. J. Exp. Zool. 286:505-512, 2000. PMID- 10684576 TI - Fundamental cryobiology of rat immature and mature oocytes: hydraulic conductivity in the presence of Me(2)SO, Me(2)SO permeability, and their activation energies. AB - The hydraulic conductivity in the presence of dimethyl sulfoxide Me(2)SO (L(p)(Me(2)SO)), Me(2)SO (P(Me(2)SO)) permeability and reflection coefficient (sigma) of immature (germinal vesicle; GV) and mature (metaphase II; MII) rat oocytes were determined at various temperatures. A temperature controlled micropipette perfusion technique was used to conduct experiments at five different temperatures (30, 20, 10, 4, and -3 degrees C). Kedem and Katchalsky membrane transport theory was used to describe the cell volume kinetics. The cell volumetric changes of oocytes were calculated from the measurement of two oocyte diameters, assuming a spherical shape. The activation energies (E(a)) of L(p)(Me(2)SO) and P(Me(2)SO) were calculated using the Arrhenius equation. Activation energies of L(p)(Me(2)SO) for GV and MII oocytes were 34.30 Kcal/mol and 16.29 Kcal/mol, respectively; while the corresponding E(a)s of P(Me(2)SO) were 19.87 Kcal/mol and 21.85 Kcal/mol, respectively. These permeability parameters were then used to calculate cell water loss in rat oocytes during cooling at subzero temperatures. Based on these values, the predicted optimal cooling rate required to maintain extra- and intracellular water in near equilibrium for rat GV stage oocytes was found to be between 0.05 degrees C/min and 0. 025; while for rat MII oocytes, the corresponding cooling rate was 1 degrees C/min. These data suggest that standard cooling rates used for mouse oocytes (e.g., 0.5-1 degrees C/min) can also be employed to cryopreserve rat MII oocytes. However, the corresponding cooling rate required to avoid damage must be significantly slower for the GV stage rat oocyte. J. Exp. Zool. 286:523-533, 2000. PMID- 10684575 TI - 1H-NMR and (31)P-NMR analysis of energy metabolism of quiescent and motile turbot (Psetta maxima) spermatozoa. AB - 31P-NMR and (1)H-NMR were used to monitor changes of several compounds with high energy bonds and metabolites prior to and after the initiation of motility of turbot spermatozoa (Psetta maxima). The obtained (31)P-NMR spectra revealed the presence of phosphomonoesters, phosphodiester, intracellular inorganic phosphate (Pi), phosphocreatine (PCr), and free nucleotide triphosphate. Following the activation of motility, the di- and tri-phosphate nucleotides, PCr, phosphomonoesters levels dropped while Pi levels increased. A significant increase of lactate was also seen at the end of the swimming phase. The compositions of seminal fluid and urine were also determined. Lipoproteins, formic acid, amino acid, and citric acid were detected in seminal fluid. Dimethyl amine, trimethylamine, and trimethylamine oxyde were found in urine. These data suggest that at least a part of the energy required during the swimming phase results from anaerobic fermentation and oxidative phosphorylation. J. Exp. Zool. 286:513-522, 2000. PMID- 10684577 TI - Effects of temperature on the deformity and sex differentiation of tilapia, Oreochromis mossambicus. AB - The effects of temperature on the deformity and sex differentiation of tilapia, Oreochromis mossambicus, were investigated. Zero- (the hatching day), 5-, and 10 day-old tilapia were respectively divided into 4 groups that were reared at 20, 24, 28, and 32 degrees C for 5 days. Percentages of deformity were significantly increased when tilapia were kept in the elevated temperatures (28 and 32 degrees C) before 5 days old during this experiment, whereas the lower temperature (20 degrees C) had no effect on the development of morphology. On the other hand, exposure to the lower temperature before 10 days old induced a high proportion of females whereas a high proportion of males was induced by the elevated temperature after 10 days old during this experiment. These results indicate that morphological development is influenced by temperature, particularly by the elevated temperature during a restricted developmental period. Both lower and elevated temperatures induce the gonadal feminization and masculinization, respectively, during its restricted developmental period. J. Exp. Zool. 286:534 537, 2000. PMID- 10684578 TI - Lethal and non-lethal responses of spermatozoa from a wide variety of vertebrates and invertebrates to lysenin, a protein from the coelomic fluid of the earthworm Eisenia foetida. AB - Lysenin, a novel protein that we isolated from the coelomic fluid of the earthworm Eisenia foetida, binds specifically to sphingomyelin (SM) among various phospholipids found in cell membranes, and causes cytolysis. The plasma membrane of mammalian spermatozoa is known to contain SM at relatively high levels and we therefore examined the effects of lysenin on the spermatozoa of various animals. Lysenin had lethal effects on spermatozoa of 5 of 33 species of invertebrates tested and on spermatozoa of 30 of 39 species of vertebrates. We postulated that plasma membranes of the spermatozoa of most invertebrates might not contain SM whereas those of most vertebrate species might contain SM. These possibilities were supported by our failure to detect SM chemically in the testes of three species of invertebrates, in none of which spermatozoa responded to lysenin. In contrast, we detected SM in the testes of all 25 vertebrate species examined, irrespective of a negative or positive response of spermatozoa to lysenin. None of the six species of Protista examined was affected by lysenin. Our survey suggests that, in general, the spermatozoa of animals can be grouped into two categories, invertebrate and vertebrate, depending on the absence or presence of SM in their plasma membrane. The incorporation of SM into spermatozoa seems first to have occurred in protochordates during the course of evolution. Discussions about the exceptional responses to lysenin observed in the spermatozoa of five species of invertebrates and of nine species of vertebrates are made from phylogenetic and reproductive viewpoints. J. Exp. Zool. 286:538-549, 2000. PMID- 10684579 TI - Evolutionary developmental genetics of floral symmetry: the revealing power of Linnaeus' monstrous flower. AB - Actinomorphic flowers have several planes of reflectional symmetry while zygomorphic flowers have just one. In a number of independent cases, actinomorphic flowers have arisen from zygomorphic ones during evolution. A famous example, studied by Linnaeus, is an actinomorphic variety of the common toadflax Linaria vulgaris. It has been shown now that this mutant carries a defect in LCYC, a homolog of the CYC gene, which controls zygomorphy in Antirrhinum majus.((1)) Interestingly, the mutant phenotype is not due to changes in the LCYC nucleotide sequence but rather to an extensive, heritable methylation of the gene.((1)) A second gene controlling zygomorphy in snapdragon, DICH, has recently also been shown to be a CYC homolog and both genes share significant sequence similarity with TB1, one of the key genes of maize domestication. The respective family of genes, probably encoding transcription factors, might thus become both a useful instrument and a target of future plant evolutionary developmental genetics. PMID- 10684580 TI - AML1 haploinsufficiency, gene dosage, and the predisposition to acute leukemia. AB - Hematopoiesis is the complex developmental process through which undifferentiated, pluripotent, hematopoietic stem cells come to generate mature, functional blood cells. This process is regulated in large part by specific transcription factors that control expression of genes necessary for the developmental sequence. Leukemias represent one form of disruption of this normal developmental process, and studies over the past few years have shown that many of the genes that underlay leukemogenesis are also essential for normal hematopoiesis. In an interesting recent example, Song et al.((1)) demonstrate that haploinsufficiency of the AML1 gene is the genetic basis of a form of familial thrombocytopenia which predisposes the affected individuals to the development of acute myeloid leukemia. Here we summarize Song's paper and current information describing the interesting dosage effects of this gene and other members of its gene family. PMID- 10684581 TI - Cell fate choices in Drosophila tracheal morphogenesis. AB - The Drosophila tracheal system is a branched tubular structure that supplies air to target tissues. The elaborate tracheal morphology is shaped by two linked inductive processes, one involving the choice of cell fates, and the other a guided cell migration. We will describe the molecular basis for these processes, and the allocation of cell fate decisions to four temporal hierarchies. First, tracheal placodes are specified within the embryonic ectoderm. Subsequently, branch fates are allocated within the tracheal placodes, prior to migration. Localized presentation of the FGF ligand, Branchless, to tracheal cells that express the FGF receptor, Breathless, guides migration. Once cell migration is initiated, distinct cell fates are determined within each migrating branch. Finally, inhibitory feedback mechanisms ensure the correct assignment of these fates. Tracheal cell fate choices are determined by signaling cascades triggered by signals emanating from the tracheal cells, as well as by ligands produced by adjacent tissues. PMID- 10684582 TI - Common structural features in gramicidin and other ion channels. AB - This review compares and contrasts the structures of several different types of ion channels with known three-dimensional structures, including gramicidin and the family of peptaibol channels, as well as the Streptomyces lividans potassium channel, to reveal common features in their structures that relate to their functional roles in ion binding and transport across membranes. Specifically, the locations of aromatic amino acids, the dimensions of the molecules, the multimeric nature of the channels and the roles of hydrogen bonds in stabilising such structures, the means by which the channels open and close, and the chemical nature of the groups which make up the channel lumen are discussed. The emphasis is on the commonality of features found in model channels, which may ultimately be found in other biological channel structures. PMID- 10684583 TI - Degradation of mRNA in bacteria: emergence of ubiquitous features. AB - The amount of a messenger RNA available for protein synthesis depends on the efficiency of its transcription and stability. The mechanisms of degradation that determine the stability of mRNAs in bacteria have been investigated extensively during the last decade and have begun to be better understood. Several endo- and exoribonucleases involved in the mRNA metabolism have been characterized as well as structural features of mRNA which account for its stability have been determined. The most important recent developments have been the discovery that the degradosome-a multiprotein complex containing an endoribonuclease (RNase E), an exoribonuclease (polynucleotide phosphorylase), and a DEAD box helicase (RhlB) has a central role in mRNA degradation and that oligo(A) tails synthesized by poly(A) polymerase facilitate the degradation of mRNAs and RNA fragments. Moreover, the phosphorylation status and the base pairing of 5' extremities, together with 3' secondary structures of transcriptional terminators, contribute to the stability of primary transcripts. Degradation of mRNAs can follow several independent pathways. Interestingly, poly(A) tails and multienzyme complexes also control the stability and the degradation of eukaryotic mRNAs. These discoveries have led to the development of refined models of mRNA degradation. PMID- 10684584 TI - Protein kinase C binding partners. AB - Members of the protein kinase C family respond to second messengers and are involved in controlling a broad array of cellular functions. The overlapping specificity and promiscuity of these proteins has promoted the view that specific binding proteins constrain individual family members to create the appropriate specificity of action. It is speculated that such protein kinase C-regulator protein interactions affect substrate availability as well as exposure to allosteric activator(s) and that consequent interactions specify cellular location and impose integration with other signaling systems. These predicted features have been realized in the identification of many protein kinase C interacting proteins and examples of these are discussed. PMID- 10684585 TI - Phagosome dynamics and function. AB - Phagocytosis of microorganisms and other particles is mediated most efficiently by receptors such as Fc-receptors (FcR) and complement-receptors (C3R). Interaction between these receptors and ligands on the particle results in signal transduction events that lead to actin polymerisation and phagosome formation. The phagosome then undergoes a maturation process whereby it transforms into a phagolysosome. Phagosome maturation depends on interactions (fusion events) with early and late endosomes as well as with lysosomes. The fusion processes are regulated by small GTP-binding proteins and other proteins that are also involved in fusion processes in the endocytic pathway. Although most phagocytosed microorganisms are killed in the lysosome, some pathogens have developed survival strategies and are able to live in the harsh conditions in the phagolysosome or interfere with the maturation process and thereby evade destruction by acid hydrolases. PMID- 10684586 TI - Notch signaling in the nervous system. Pieces still missing from the puzzle. AB - Notch has been known for many years as a receptor for inhibitory signals that shapes the pattern of the nervous system during its development. Genes in the Notch pathway function to prevent neural determination so that only a subset of the available ectodermal cells become neural precursors. The localization of Notch signaling is crucial for determining where neural precursor cells arise on a cell-by-cell basis. The unresolved problem is that studies of the expression of Notch protein and its ligands are inconsistent with the pattern of neurogenesis. During neural cell fate specification, distributions of Notch protein and of its ligand Delta appear uniform. Under the reigning paradigm, such widespread expression should lead to N signal transduction in all cells and thereby prevent any neural specification. Yet, contrary to this expectation, neural elements still form, in characteristic patterns, hence, Notch signal transduction must have been inactive in the precursor cells. The mechanism preventing Notch signaling in certain cells must be posttranslational but it has not yet been identified. This review will outline the experimental evidence supporting this view of Notch signaling, and briefly evaluate some of the possible mechanisms that have been suggested. PMID- 10684587 TI - A new functional classification of tumor-suppressing genes and its therapeutic implications. AB - Cell fusion studies have demonstrated that malignancy can be suppressed by a single dose of malignancy suppressor genes (MSGs), indicating that malignancy is a recessive phenotype. Correspondingly, it is widely believed that mutational inactivation of both alleles of tumor suppressor genes (TSGs), in familial and sporadic tumors, is the formal proof of the recessive nature of malignancy. Evidence presented here, however, shows that unlike MSGs, identified solely through cell fusion studies with no gene of this class yet cloned, many well known TSGs have gene dosage effects and inhibit cellular growth in vitro. Moreover, homozygous inactivation of a growth-inhibitory TSG (GITSG) is not directly correlated with malignancy. An alternative interpretation is provided for the loss of wild-type alleles of these genes in the tumors. It is concluded that the MSGs and the GITSGs do not belong to the same class of genes. The functional classification of tumor-suppressing genes has important implications for developing effective cancer therapies. PMID- 10684588 TI - Bystander effects: a concept in need of clarification. AB - An increasing body of evidence indicates that the response to genotoxic agents such as radiation or drugs is a group phenomenon, rather than the summed response of individual independent cells to injury. Thus, a complex contagion-like response may spread beyond the initial impact of an agent to enlarge its effect. This indirect effect, termed "Bystander Effect," is multifaceted and may play a significant role in the therapy of tumors and in carcinogenesis. A better understanding of this phenomenon is needed in order to modulate treatment protocols to therapeutic advantage and to provide more rational guidelines for the evaluation of environmental hazards. PMID- 10684589 TI - The coral Acropora: what it can contribute to our knowledge of metazoan evolution and the evolution of developmental processes. AB - The diploblastic Cnidaria form one of the most ancient metazoan phyla and thus provide a useful outgroup for comparative studies of the molecular control of development in the more complex, and more often studied, triploblasts. Among cnidarians, the reef building coral Acropora is a particularly appropriate choice for study. Acropora belongs to the Anthozoa, which several lines of evidence now indicate is the basal class within the phylum Cnidaria, and has the practical advantages that its reproduction is predictable, external and accessible and that the base content of its genome is not strongly biased. The Acropora system has already provided insights into ancestral linkages of homeobox genes and the evolution of the Pax genes, and has the potential to provide further new perspectives on the age, role in development, and evolution of these and other gene families. PMID- 10684590 TI - What transgenic mice tell us about neurodegenerative disease. AB - The recent broad advance in our understanding of human neurodegenerative diseases is based on the application of a new molecular approach. Through linkage analysis, the genes responsible for Huntington's disease, the spinocerebellar ataxias, and familial forms of Alzheimer's disease and amyotrophic lateral sclerosis (ALS) have been identified and cloned. The characterization of pathogenic mutations in such genes allows the creation of informative transgenic mouse models as, without exception, the genetic forms of adult neurodegenerative disease are due to toxicity of the mutant protein. Transgenic models provide insight into the oxidative mechanisms in ALS pathogenesis, the pathogenicity of expanded polyglutamine tracts in CAG triplet repeat disorders, and amyloidogenesis in Alzheimer's disease. Although such models have their limitations, they currently provide the best entry point for the study of human neurodegenerative diseases. PMID- 10684591 TI - Drosophila: flying high in Zurich. PMID- 10684592 TI - Popper. PMID- 10684593 TI - The utility of Popper's philosophy in biology. PMID- 10684594 TI - Don't belittle Popper. Refutation cannot be refuted in biology, either. PMID- 10684595 TI - Kinetic studies of the mechanism of carbon-hydrogen bond breakage by the heterotetrameric sarcosine oxidase of Arthrobacter sp. 1-IN. AB - The reaction of heterotetrameric sarcosine oxidase (TSOX) of Arthrobactor sp. 1 IN has been studied by stopped-flow spectroscopy, with particular emphasis on the reduction of the enzyme by sarcosine. Expression of the cloned gene encoding TSOX in Escherichia coli enables the production of TSOX on a scale suitable for stopped-flow studies. Treatment of the enzyme with sulfite provides the means for selective formation of a flavin-sulfite adduct with the covalent 8alpha-(N(3) histidyl)-FMN. Formation of the sulfite-flavin adduct suppresses internal electron transfer between the noncovalent FAD (site of sarcosine oxidation) and the covalent FMN (site of enzyme oxidation) and thus enables detailed characterization of the kinetics of FAD reduction by sarcosine using stopped-flow methods. The rate of FAD reduction displays a simple hyperbolic dependence on sarcosine concentration. Studies in the pH range 6.5-10 indicate there are no kinetically influential ionizations in the enzyme-substrate complex. A plot of the limiting rate of flavin reduction/the enzyme-substrate dissociation constant (k(lim)/K(d)) versus pH is bell-shaped and characterized by two macroscopic pK(a) values of 7.4 +/- 0.1 and 10.4 +/- 0.2: potential candidates for the two ionizable groups are discussed with reference to the structure of monomeric sarcosine oxidase (MSOX). The kinetic data are discussed with reference to potential mechanisms for the oxidation of amine molecules by flavoenzymes. Additionally, kinetic isotope effect studies of the rate of C-H bond breakage suggest that a ground-state quantum tunneling mechanism for H-transfer, facilitated by the low-frequency thermal motions of the protein molecule, accounts for C-H bond cleavage by TSOX. TSOX thus provides another example of C-H bond breakage by ground-state quantum tunneling, driven by protein dynamics [vibrationally enhanced ground-state quantum tunneling (VEGST)], for the oxidation of amines by enzymes. PMID- 10684596 TI - Reactions of Pseudomonas 7A glutaminase-asparaginase with diazo analogues of glutamine and asparagine result in unexpected covalent inhibitions and suggests an unusual catalytic triad Thr-Tyr-Glu. AB - Pseudomonas 7A glutaminase-asparaginase (PGA) catalyzes the hydrolysis of D and L isomers of glutamine and asparagine. Crystals of PGA were reacted with diazo analogues of glutamine (6-diazo-5-oxo-L-norleucine, DON) and asparagine (5-diazo 4-oxo-L-norvaline, DONV), which are known inhibitors of the enzyme. The derivatized crystals remained isomorphous to native PGA crystals. Their structures were refined to crystallographic R = 0.20 and R(free) = 0.24 for PGA DON and R = 0.19 and R = 0.23 for PGA-DONV. Difference Fourier electron density maps clearly showed that both DON and DONV inactivate PGA through covalent inhibition. Continuous electron density connecting the inhibitor to both Thr20 and Tyr34 of the flexible loop was observed providing strong evidence that Thr20 is the primary catalytic nucleophile and that Tyr34 plays an important role in catalysis as well. The unexpected covalent binding observed in the PGA-DON and PGA-DONV complexes shows that a secondary reaction involving the formation of a Tyr34-inhibitor bond takes place with concomitant inactivation of PGA. The predicted covalent linkage is not seen, however, suggesting an alternative method of inhibition not yet seen for these diazo analogues. These surprising results give insight as to the role of the flexible loop Thr and Tyr in the catalytic mechanism. PMID- 10684597 TI - Mutations at the histidine 249 ligand profoundly alter the spectral and iron binding properties of human serum transferrin N-lobe. AB - Human serum transferrin is an iron-binding and -transport protein which carries iron from the blood stream into various cells. Iron is held in two deep clefts located in the N- and C-lobes by coordinating to four amino acid ligands, Asp 63, Tyr 95, Tyr 188, and His 249 (N-lobe numbering), and to two oxygens from carbonate. We have previously reported the effect on the iron-binding properties of the N-lobe following mutation of the ligands Asp 63, Tyr 95, and Tyr 188. Here we report the profound functional changes which result from mutating His 249 to Ala, Glu, or Gln. The results are consistent with studies done in lactoferrin which showed that the histidine ligand is critical for the stability of the iron binding site [H. Nicholson, B. F. Anderson, T. Bland, S. C. Shewry, J. W. Tweedie, and E. N. Baker (1997) Biochemistry 36, 341-346]. In the mutant H249A, the histidine ligand is disabled, resulting in a dramatic reduction in the kinetic stability of the protein toward loss of iron. The H249E mutant releases iron three times faster than wild-type protein but shows significant changes in both EPR spectra and the binding of anion. This appears to be the net effect of the metal ligand substitution from a neutral histidine residue to a negative glutamate residue and the disruption of the "dilysine trigger" [MacGillivray, R. T. A., Bewley, M. C., Smith, C. A., He, Q.-Y., Mason, A. B., Woodworth, R. C., and Baker, E. N. (2000) Biochemistry 39, 1211-1216]. In the H249Q mutant, Gln 249 appears not to directly contact the iron, given the similarity in the spectroscopic properties and the lability of iron release of this mutant to the H249A mutant. Further evidence for this idea is provided by the preference of both the H249A and H249Q mutants for nitrilotriacetate rather than carbonate in binding iron, probably because NTA is able to provide a third ligation partner. An intermediate species has been identified during the kinetic interconversion between the NTA and carbonate complexes of the H249A mutant. Thus, mutation of the His 249 residue does not abolish iron binding to the transferrin N-lobe but leads to the appearance of novel iron-binding sites of varying structure and stability. PMID- 10684598 TI - Mutation of the iron ligand His 249 to Glu in the N-lobe of human transferrin abolishes the dilysine "trigger" but does not significantly affect iron release. AB - Serum transferrin is the major iron transport protein in humans. Its function depends on its ability to bind iron with very high affinity, yet to release this bound iron at the lower intracellular pH. Possible explanations for the release of iron from transferrin at low pH include protonation of a histidine ligand and the existence of a pH-sensitive "trigger" involving a hydrogen-bonded pair of lysines in the N-lobe of transferrin. We have determined the crystal structure of the His249Glu mutant of the N-lobe half-molecule of human transferrin and compared its iron-binding properties with those of the wild-type protein and other mutants. The crystal structure, determined at 2.4 A resolution (R-factor 19.8%, R(free) 29.4%), shows that Glu 249 is directly bound to iron, in place of the His ligand, and that a local movement of Lys 296 has broken the dilysine interaction. Despite the loss of this potentially pH-sensitive interaction, the H249E mutant is only slightly more acid-stable than wild-type and releases iron slightly faster. We conclude that the loss of the dilysine interaction does make the protein more acid stable but that this is counterbalanced by the replacement of a neutral ligand (His) by a negatively charged one (Glu), thus disrupting the electroneutrality of the binding site. PMID- 10684599 TI - Camelid heavy-chain variable domains provide efficient combining sites to haptens. AB - Camelids can produce antibodies devoid of light chains and CH1 domains (Hamers Casterman, C. et al. (1993) Nature 363, 446-448). Camelid heavy-chain variable domains (VHH) have high affinities for protein antigens and the structures of two of these complexes have been determined (Desmyter, A. et al. (1996) Nature Struc. Biol. 3, 803-811; Decanniere, K. et al. (1999) Structure 7, 361-370). However, the small size of these VHHs and their monomeric nature bring into question their capacity to bind haptens. Here, we have successfully raised llama antibodies against the hapten azo-dye Reactive Red (RR6) and determined the crystal structure of the complex between a dimer of this hapten and a VHH fragment. The surface of interaction between the VHH and the dimeric hapten is large, with an area of ca. 300 A(2); this correlates well with the low-dissociation constant of 22 nM measured for the monomer. The VHH fragment provides an efficient combining site to the RR6, using its three CDR loops. In particular, CDR1 provides a strong interaction to the hapten through two histidine residues bound to its copper atoms. VHH fragments might, therefore, prove to be valuable tools for selecting, removing, or capturing haptens. They are likely to play a role in biotechnology extending beyond protein recognition alone. PMID- 10684600 TI - Mutagenesis of E477 or K505 in the B' domain of human topoisomerase II beta increases the requirement for magnesium ions during strand passage. AB - A type II topoisomerase is essential for decatenating DNA replication products, and it accomplishes this task by passing one DNA duplex through a transient break in a second duplex. The B' domain of topoisomerase II contains three highly conserved motifs, EGDSA, PL(R/K)GK(I/L/M)LNVR, and IMTD(Q/A)DXD. We have investigated these motifs in topoisomerase II beta by mutagenesis, and report that they play a critical role in establishing the DNA cleavage-religation equilibrium. In addition, the mutations E477Q (EGDSA) and K505E (PLRGKILNVR) increase the optimal magnesium ion concentration for strand passage, without affecting the Mg(2+) dependence of ATP hydrolysis. It is likely that the binding affinity of the magnesium ion(s) specifically required for DNA cleavage has been reduced by these mutations. The crystal structure of yeast topo II indicates that residues E477 and K505 may help to position the three aspartate residues of the IMTD(Q/A)DXD motif for magnesium ion coordination, and we propose two possible locations for the magnesium ion binding site(s). These observations are consistent with a previous model in which the B' domain is positioned such that these acidic residues lie next to the active site tyrosine residue. A magnesium ion bound by these aspartate residues could therefore mediate the DNA cleavage religation reaction. PMID- 10684601 TI - Structural and mechanistic basis for the activation of a low-molecular weight protein tyrosine phosphatase by adenine. AB - Although the activation of low-molecular weight protein tyrosine phosphatases by certain purines and purine derivatives was first described three decades ago, the mechanism of this rate enhancement was unknown. As an example, adenine activates the yeast low-molecular weight protein tyrosine phosphatase LTP1 more than 30 fold. To examine the structural and mechanistic basis of this phenomenon, we have determined the crystal structure of yeast LTP1 complexed with adenine. In the crystal structure, an adenine molecule is found bound in the active site cavity, sandwiched between the side chains of two large hydrophobic residues at the active site. Hydrogen bonding to the side chains of other active site residues, as well as some water-mediated hydrogen bonds, also helps to fix the position of the bound adenine molecule. An ordered water was found in proximity to the bound phosphate ion present in the active site, held by hydrogen bonding to N3 of adenine and Odelta1 of Asp-132. On the basis of the crystal structure, we propose that this water molecule is the nucleophile that participates in the dephosphorylation of the phosphoenzyme intermediate. Solvent isotope effect studies show that there is no rate-determining transfer of a solvent-derived proton in the transition state for the dephosphorylation of the phosphoenzyme intermediate. Such an absence of general base catalysis of water attack is consistent with the stability of the leaving group, namely, the thiolate anion of Cys-13. Consequently, adenine activates the enzyme by binding and orienting a water nucleophile in proximity to the phosphoryl group of the phosphoenzyme intermediate, thus increasing the rate of the dephosphorylation step, a step that is normally the rate-limiting step of this enzymatic reaction. PMID- 10684602 TI - Residues in Cdc42 that specify binding to individual CRIB effector proteins. AB - Cdc42 is a member of the Rho family of small G proteins. Signal transduction events emanating from Cdc42 lead to cytoskeletal rearrangements, cell proliferation, and cell differentiation. Many effector proteins have been identified for Cdc42; however, it is not clear how certain effectors specifically recognize and bind to Cdc42, as opposed to Rac or Rho, or in many cases, which effector controls what cellular events. Mutations were introduced into Cdc42 at residues: Met1, Val8, Phe28, Tyr32, Val33, Thr35, Val36, Phe37, Asp38, Tyr40, Val42, Met45, Ile46, Glu127, Ala130, Asn132, Gln134, Lys135, and Leu174. Measurements were made of their equilibrium binding constants to the Cdc42 binding domains of the CRIB effectors ACK, PAK, and WASP and to the GTPase activating protein Rho GAP. Generally, mutations in the effector loop have an equally deleterious effect on binding to all CRIB proteins tested, though the F37A mutation resulted in significant selectivity. Residues outside the effector loop were found to be important for binding of Cdc42 to CRIB containing proteins and also to contribute to selectivity. Mutations such as V42A and L174A resulted in large, selective changes in binding to specific CRIB effectors. Neither mutation resulted in alteration in PAK binding, whereas both severely disrupt binding to ACK and only L174A disrupted binding to WASP. These mutations are interpreted using the structures of the Cdc42/ACK and Cdc42/WASP complexes to give insight into how effectors can specifically recognize Cdc42. Those mutations in Cdc42 that inhibit certain interactions, while retaining others, should aid investigations of the role of specific effectors in Cdc42 signaling in vivo. PMID- 10684603 TI - Contribution of surface salt bridges to protein stability. AB - The role of surface salt bridges in protein stabilization has been a source of controversy. Here we present the NMR structure of a hyperthermophilic rubredoxin variant (PFRD-XC4) and the thermodynamic analysis of two surface salt bridges by double mutant cycles. This analysis shows that the surface side chain to side chain salt bridge between Lys 6 and Glu 49 does not stabilize PFRD-XC4. The main chain to side chain salt bridge between the N-terminus and Glu 14 was, however, found to stabilize PFRD-XC4 by 1. 5 kcal mol(-)(1). The entropic cost of making a surface salt bridge involving the protein's backbone is reduced, since the backbone has already been immobilized upon protein folding. PMID- 10684604 TI - Actinonin, a naturally occurring antibacterial agent, is a potent deformylase inhibitor. AB - Peptide deformylase (PDF) is essential in prokaryotes and absent in mammalian cells, thus making it an attractive target for the discovery of novel antibiotics. We have identified actinonin, a naturally occurring antibacterial agent, as a potent PDF inhibitor. The dissociation constant for this compound was 0.3 x 10(-)(9) M against Ni-PDF from Escherichia coli; the PDF from Staphylococcus aureus gave a similar value. Microbiological evaluation revealed that actinonin is a bacteriostatic agent with activity against Gram-positive and fastidious Gram-negative microorganisms. The PDF gene, def, was placed under control of P(BAD) in E. coli tolC, permitting regulation of PDF expression levels in the cell by varying the external arabinose concentration. The susceptibility of this strain to actinonin increases with decreased levels of PDF expression, indicating that actinonin inhibits bacterial growth by targeting this enzyme. Actinonin provides an excellent starting point from which to derive a more potent PDF inhibitor that has a broader spectrum of antibacterial activity. PMID- 10684605 TI - Structure of a NifS homologue: X-ray structure analysis of CsdB, an Escherichia coli counterpart of mammalian selenocysteine lyase. AB - Escherichia coli CsdB, a NifS homologue with a high specificity for L selenocysteine, is a pyridoxal 5'-phosphate (PLP)-dependent dimeric enzyme that belongs to aminotransferases class V in fold-type I of PLP enzymes and catalyzes the decomposition of L-selenocysteine into selenium and L-alanine. The crystal structure of the enzyme has been determined by the X-ray crystallographic method of multiple isomorphous replacement and refined to an R-factor of 18.7% at 2.8 A resolution. The subunit structure consists of three parts: a large domain of an alpha/beta-fold containing a seven-stranded beta-sheet flanked by seven helices, a small domain containing a four-stranded antiparallel beta-sheet flanked by three alpha-helices, and an N-terminal segment containing two alpha-helices. The overall fold of the subunit is similar to those of the enzymes belonging to the fold-type I family represented by aspartate aminotransferase. However, CsdB has several structural features that are not observed in other families of the enzymes. A remarkable feature is that an alpha-helix in the lobe extending from the small domain to the large domain in one subunit of the dimer interacts with a beta-hairpin loop protruding from the large domain of the other subunit. The extended lobe and the protruded beta-hairpin loop form one side of a limb of each active site in the enzyme. The most striking structural feature of CsdB lies in the location of a putative catalytic residue; the side chain of Cys364 on the extended lobe of one subunit is close enough to interact with the gamma-atom of a modeled substrate in the active site of the subunit. Moreover, His55 from the other subunit is positioned so that it interacts with the gamma- or beta-atom of the substrate and may be involved in the catalytic reaction. This is the first report on three-dimensional structures of NifS homologues. PMID- 10684606 TI - Channeling of carbon monoxide during anaerobic carbon dioxide fixation. AB - Carbon monoxide is an intermediate in carbon dioxide fixation by diverse microbes that inhabit anaerobic environments including the human colon. These organisms fix CO(2) by the Wood-Ljungdahl pathway of acetyl-CoA biosynthesis. The bifunctional CO dehydrogenase/acetyl-CoA synthase (CODH/ACS) catalyzes several key steps in this pathway. CO(2) is reduced to CO at a nickel iron-sulfur cluster called cluster C located in the CODH subunit. Then, CO is condensed with a methyl group and coenzyme A at cluster A, another nickel iron-sulfur cluster in the ACS subunit. Spectroscopic studies indicate that clusters A and C are at least 10-15 A apart. To gain a better understanding of how CO production and utilization are coordinated, we have studied an isotopic exchange reaction between labeled CO(2) and the carbonyl group of acetyl-CoA with the CODH/ACS from Clostridium thermoaceticum. When solution CO is provided at saturating levels, only CO(2) derived CO is incorporated into the carbonyl group of acetyl-CoA. Furthermore, when high levels of hemoglobin or myoglobin are added to remove CO from solution, there is only partial inhibition of the incorporation of CO(2)-derived CO into acetyl-CoA. These results provide strong evidence for the existence of a CO channel between cluster C in the CODH subunit and cluster A in the ACS subunit. The existence of such a channel would tightly couple CO production and utilization and help explain why high levels of this toxic gas do not escape into the environment. Instead, microbes sequester this energy-rich carbon source for metabolic reactions. PMID- 10684607 TI - Probes of a role for remote binding interactions on hydrogen tunneling in the horse liver alcohol dehydrogenase reaction. AB - A tunneling contribution to hydride transfer has been demonstrated previously in the oxidation of benzyl alcohol catalyzed by an active-site mutant (F93W) of horse liver alcohol dehydrogenase (LADH) [Bahnson, B. J., et al. (1993) Biochemistry 32, 5503-5507]. Mutation of a residue that lies directly behind the nicotinamide ring of the bound cofactor has further shown that side-chain bulk can contribute to catalytic efficiency and tunneling in a correlated fashion [Bahnson, B. J., et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 12797-12802]. Second site mutations of F93W have now been made at positions more remote from the active site. In particular, we have focused on an isoleucine residue that interacts with the adenine moiety of the NAD(+) cofactor, 20 A from the nicotinamide ring. Replacement of this remote residue with glycine (F93W:I224G), alanine (F93W:I224A), valine (F93W:I224V), and leucine (F93W:I224L) is concluded to destabilize the binding of NAD(+). All double mutants exhibited a K(M) for NAD(+) that is 2-25 times higher than that for the F93W enzyme. However, neither the catalytic efficiency for turnover of benzyl alcohol [k(cat)/K(M(benzyl alcohol))] nor the relationship between the secondary k(H)/k(T) and k(D)/k(T) isotope effects for benzyl alcohol oxidation was significantly affected. The lack of differences observed in the isotope effects indicates that these mutations have little effect on the extent of hydrogen tunneling in the reaction. The complete removal of the side chain at position 224 in the F93W:I224G enzyme resulted in a less than 5% decrease in the ratio of the secondary isotope effects, maintaining the ratio above the semiclassical limit for the indication of tunneling in the reaction. By contrast, K(i) for NAD(+) increased 60-fold for this mutant. The results obtained with F93W:I224G are consistent with remote interactions that affect the association and binding of cofactor in a reactive conformation. However, once this conformation is achieved, hydride transfer and its tunneling component proceed as with the single F93W mutant enzyme, uninfluenced by the remote mutation. Replacement of other side chains, with alpha carbon positions from about 8 to over 20 A from the C4 position of the nicotinamide ring, demonstrated a similar insensitivity of k(cat)/K(M(benzyl alcohol)) to protein modification. Comparison to earlier studies with active-site mutants of LADH implicates a role for proximal, but not distal, side chains in the modulation of hydrogen tunneling for this enzyme. PMID- 10684608 TI - Mechanistic analysis of the argE-encoded N-acetylornithine deacetylase. AB - The E. coli argE-encoded N-acetyl-L-ornithine deacetylase has been cloned, expressed, and purified in high yield. The substrate specificity of the enzyme is relatively broad, with a number of alpha-N-acyl-L-amino acids exhibiting activity, including both alpha-N-acetyl- and alpha-N-formylmethionine that exhibit higher activity than alpha-N-acetyl-L-ornithine. Sequence homolgy suggests that the enzyme is a member of the metal-dependent aminoacylase family, and the purified enzyme contains a single atom of zinc per monomer. The activity of this enzyme can be increased greater than 2-fold by the addition of zinc, or 8 fold by the addition of cobalt. This suggests that the enzyme can accommodate two metal ions at the active site. The pH dependence of the kinetic parameters has been determined and revealed the presence of two enzymic groups, one functioning as a general base and one functioning as a general acid. Solvent kinetic isotope effects on the hydrolysis of N-acetylornithine have been determined, and a linear proton inventory suggests that a single proton transfer occurs in a partially rate-limiting step. A chemical mechanism is proposed and compared with other mechanisms determined for other members of the aminoacylase family. PMID- 10684609 TI - Characterization of five catalytic activities associated with the NADPH:2 ketopropyl-coenzyme M [2-(2-ketopropylthio)ethanesulfonate] oxidoreductase/carboxylase of the Xanthobacter strain Py2 epoxide carboxylase system. AB - The bacterial metabolism of propylene proceeds by epoxidation to epoxypropane followed by carboxylation to acetoacetate. Epoxypropane carboxylation is a minimetabolic pathway that requires four enzymes, NADPH, NAD(+), and coenzyme M (CoM; 2-mercaptoethanesulfonate) and occurs with the overall reaction stoichiometry: epoxypropane + CO(2) + NADPH + NAD(+) + CoM --> acetoacetate + H(+) + NADP(+) + NADH + CoM. The terminal enzyme of the pathway is NADPH:2 ketopropyl-CoM [2-(2-ketopropylthio)ethanesulfonate] oxidoreductase/carboxylase (2-KPCC), an FAD-containing enzyme that is a member of the NADPH:disulfide oxidoreductase family of enzymes and that catalyzes the reductive cleavage and carboxylation of 2-ketopropyl-CoM to form acetoacetate and CoM according to the reaction: 2-ketopropyl-CoM + NADPH + CO(2) --> acetoacetate + NADP(+) + CoM. In the present work, 2-KPCC has been characterized with respect to the above reaction and four newly discovered partial reactions of relevance to the catalytic mechanism, and each of which requires the formation of a stabilized enolacetone intermediate. These four reactions are (1) NADPH-dependent cleavage and protonation of 2-ketopropyl-CoM to form NADP(+), CoM, and acetone, a reaction analogous to the physiological reaction but in which H(+) is the electrophile; (2) NADP(+)-dependent synthesis of 2-ketopropyl-CoM from CoM and acetoacetate, the reverse of the physiologically important forward reaction; (3) acetoacetate decarboxylation to form acetone and CO(2); and (4) acetoacetate/(14)CO(2) exchange to form (14)C(1)-acetoacetate and CO(2). Acetoacetate decarboxylation and (14)CO(2) exchange occurred independent of NADP(H) and CoM, demonstrating that these substrates are not central to the mechanism of enolate generation and stabilization. 2-KPCC did not uncouple NADPH oxidation or NADP(+) reduction from the reactions involving cleavage or formation of 2-ketopropyl-CoM. N Ethylmaleimide inactivated the reactions forming/using 2-ketopropyl-CoM but did not inactivate acetoacetate decarboxylation or (14)CO(2) exchange reactions. The biochemical characterization of 2-KPCC and the associated five catalytic activities has allowed the formulation of an unprecedented mechanism of substrate activation and carboxylation that involves NADPH oxidation, a redox active disulfide, thiol-mediated reductive cleavage of a C-S thioether bond, the formation of a CoM:cysteine mixed disulfide, and enolacetone stabilization. PMID- 10684610 TI - Stepwise modulation of ATPase activity, nucleotide trapping, and sliding motility of myosin S1 by modification of the thiol region with residues of increasing size. AB - Rabbit muscle myosin S1 was modified either at SH1 alone or at both SH1 and SH2, using a series of alkylthiolating reagents of increasing size, designed for correlating gradually changing structural disturbances in the thiol region with functional impairments in the myosin head. The reagents were of the type H(CH(2))(n)()-S-NTB, (NTB = 2-nitro-5-thiobenzoate) (n = 1, 2, 5, 8, 9, 10, 11, and 12). Modification of only SH1 led to the expected activation of the Ca(2+) ATPase, but only with small reagents, while reagents with n > or = 10 caused inhibition of the Ca(2+)-ATPase. Modification of both SH1 and SH2 showed the expected inhibition of Ca(2+)-ATPase but likewise allowed considerable residual Ca(2+)-ATPase activity if the residues were small. Trapping of the nucleotide, known to occur with cross-linking reagents, was seen also with monovalent reagents, provided their length exceeded n = 9 or 10. All S1 derivatives prepared in this study possessed an affinity for actin comparable to native S1 but lacked sliding motility in in vitro motility assays. The biochemical data of this study can be related to existing models of myosin S1 and recent structural data [Houdusse, A., Kalabokis, V. N., Himmel, D., Szent-Gyorgyi, A. G., and Cohen, C. (1999) Cell 97, 459-470] by making the assumptions that modification at SH1 prevents the formation of the SH1 helix mandatory for the transmission of conformational energy and that mobility of the thiol region is a prerequisite for ATPase activity. Immobilization of the thiol region by residues of increasing size apparently leads to lower enzyme activity and, finally, to inhibition of nucleotide exchange. PMID- 10684611 TI - Novel interaction of the voltage-dependent sodium channel (VDSC) with calmodulin: does VDSC acquire calmodulin-mediated Ca2+-sensitivity? AB - The voltage-dependent sodium channel (VDSC) interacts with intracellular molecules to modulate channel properties and localizations in neuronal cells. To study protein interactions, we applied yeast two-hybrid screening to the cytoplasmic C-terminal domain of the main pore-forming alpha-subunit. We found a novel interaction between the C-terminal domain and calmodulin (CaM). By two hybrid interaction assays, we specified the interaction site of VDSC in a C terminal region, which is composed of 38 amino acid residues and contains both IQ like and Baa motifs. Using a fusion protein of the C-terminal domain, we showed that interaction with CaM occurred in the presence and absence of Ca(2+). Two synthetic peptides, each covering the IQ-like (NaIQ) or the Baa motifs (NaBaa), were used to examine the binding property by a gel mobility shift assay. Although the NaIQ and NaBaa sequences are overlapped, NaBaa binds only to Ca(2+)-bound Ca(2+)CaM, whereas NaIQ binds to both Ca(2+)CaM and Ca(2+)-free apoCaM. Fluorescence spectroscopy of dansylated CaM showed Ca(2+)-dependent spectral changes not only for NaBaa.CaM but also for NaIQ.CaM. The results, taken together with other results, indicate that whereas the NaBaa.CaM complex is formed in a Ca(2+)-dependent manner, the NaIQ.CaM complex has two conformational states, distinct with respect to the peptide binding site and the CaM conformation, depending on the Ca(2+) concentration. These observations suggest the possibility that VDSC is functionally modulated through the direct CaM interaction and the Ca(2+)-dependent conformational transition of the complex. PMID- 10684612 TI - A230Y mutation of actin on subdomain 4 is sufficient for higher calcium activation of actin-activated myosin adenosinetriphosphatase in the presence of tropomyosin-troponin. AB - To probe the mechanism by which Ca(2+) activates muscle contraction through tropomyosin and troponin, we have produced mutant actins using Dictyostelium discoideum. We focused on the sequence 228-232 (QTAAS) that is located in subdomain 4 of actin, because the chimera actin in which this sequence was replaced by KAYKE showed not only poorer tropomyosin binding but also the unexpected "higher Ca(2+) activation" [Saeki, K., et al. (1996) Biochemistry 35, 14465-14472]. We found that this higher Ca(2+) activation is solely due to the A230Y mutagenesis. Because A230Y mutant actin showed normal tropomyosin binding, the higher Ca(2+) activation is not the consequence of poorer tropomyosin binding. The significance of these results is discussed in view of a three-state model [McKillop, D. F., Geeves, M. A. (1993) Biophys. J. 65, 693-701]. PMID- 10684613 TI - The carbonyl group of glutamic acid-795 is essential for gastric H+,K+-ATPase activity. AB - To study the role of Glu795offresent in the fifth transmembrane domain of the alpha-subunit of gastric H+,K+-ATPase, several mutants were generated and expressed in Sf9 insect cells. The E795Q mutant had rather similar properties as the wild-type enzyme. The apparent affinity for K+ in both the ATPase reaction and the dephosphorylation of the phosphorylated intermediate was even slightly enhanced. This indicates that the carbonyl group of Glu795 is sufficient for enzymatic activity. This carbonyl group, however, has to be at a particular position with respect to the other liganding groups, since the E795D and E795N mutants showed a strongly reduced ATPase activity, a lowered apparent K+ affinity, and a decreased steady-state phosphorylation level. In the absence of a carbonyl residue at position 795, the K+ sensitivity was either strongly decreased (E795A) or completely absent (E795L). The mutant E795L, however, showed a SCH 28080 sensitive ATPase activity in the absence of K+, as well as an enhanced spontaneous dephosphorylation rate, that could not be further enhanced by K+, suggesting that this mutant mimicks the filled K+ binding pocket. The results indicate that the Glu795 residue is involved in K+-stimulated ATPase activity and K+-induced dephosphorylation of the phosphorylated intermediate. Glu795 might also be involved in H+ binding during the phosphorylation step, since the mutants E795N, E795D, and E795A showed a decrease in the phosphorylation rate as well as in the apparent ATP affinity in the phosphorylation reaction. This indicates that Glu795 is not only involved in K+ but might also play a role in H+ binding. PMID- 10684614 TI - Active-site-directed photolabeling of the melibiose permease of Escherichia coli. AB - Covalent photolabeling of the melibiose permease (MelB) of Escherichia coli has been undertaken with the sugar analogue [(3)H]-p-azidophenyl alpha-D galactopyranoside ([(3)H]-alpha-PAPG) with the purpose of identifying the domains forming the MelB sugar-binding site. We show that alpha-PAPG is a high-affinity substrate of MelB (K(d) = 1 x 10(-)(6) M). Its binding to or transport by MelB is Na-dependent and is competitively prevented by melibiose or by the high-affinity ligand p-nitrophenyl alpha-D-galactopyranoside (alpha-NPG). Membrane vesicles containing overexpressed histidine-tagged recombinant MelB were photolabeled in the presence of [(3)H]-alpha-PAPG by irradiation with UV light (lambda = 250 nm). Eighty-five percent of the radioactivity covalently associated with the vesicles was incorporated in a polypeptide corresponding to MelB monomer. MelB labeling was completely prevented by an excess of melibiose or alpha-NPG during the assay. Radioactivity analysis of CNBr cleavage or limited proteolysis products of the purified [(3)H]-alpha-PAPG-labeled transporter suggests that several domains of MelB are targets for labeling. One of the labeled CNBr cleavage products is a peptide with an apparent molecular mass of 5.5 kDa. It is shown that (i) its amino acid sequence is that of the Asp124-Met181 domain of MelB (7.5 kDa), which includes the cytoplasmic loop 4-5 connecting helices IV and V, the hydrophobic helix V, and the outer loop connecting helices V-VI, and (ii) that Arg141 in loop 4-5 is the only labeled amino acid of this peptide. Labeling of loop 4-5 provides independent evidence that this specific domain plays a significant role in MelB transport. Comparison with the well-characterized equivalent domain of LacY suggests that sugar transporters with similar structure and substrate specificity may have conserved domains involved in sugar recognition. PMID- 10684615 TI - Moving a microtubule may require two heads: a kinetic investigation of monomeric Ncd. AB - Ncd is a minus-end-directed microtubule motor and a member of the kinesin superfamily. The Ncd dimer contains two motor domains, and cooperative interactions between the heads influence the interactions of each respective motor domain with the microtubule. The approach we have taken to understand the cooperativity between the two motor domains is to analyze the ATPase cycle of dimeric MC1 and monomeric MC6. The steps in the ATPase cycle where cooperativity occurs can be identified by comparing the two mechanisms. The rate-limiting step in the MC6 mechanism is ADP release at 3.4 s(-)(1). The observed rate constant for ATP-induced dissociation from the microtubule is 14 s(-)(1). However, the relative amplitude associated with MC6 dissociation is extremely small in comparison to the amplitude associated with dimeric MC1 dissociation kinetics. The amplitude data indicate that monomeric MC6 does not detach from the microtubule during the initial turnovers of ATP, and ATP hydrolysis is uncoupled from movement. The results show that cooperative interactions between the motor domains of the dimer are required for ATP-dependent dissociation; therefore, one function of the partner motor domain may be to weaken the interaction of the adjacent head with the microtubule. PMID- 10684616 TI - Functional properties of the heme propionates in cytochrome c oxidase from Paracoccus denitrificans. Evidence from FTIR difference spectroscopy and site directed mutagenesis. AB - By specific (13)C labeling of the heme propionates, four bands in the reduced minus-oxidized FTIR difference spectrum of cytochrome c oxidase from Paracoccus denitrificans have been assigned to the heme propionates [Behr, J., Hellwig, P., Mantele, W., and Michel, H. (1998) Biochemistry 37, 7400-7406]. To attribute these signals to the individual propionates, we have constructed seven cytochrome coxidase variants using site-directed mutagenesis of subunit I. The mutant enzymes W87Y, W87F, W164F, H403A, Y406F, R473K, and R474K were characterized by measurement of enzymatic turnover, proton pumping activity, and Vis and FTIR spectroscopy. Whereas the mutant enzymes W164F and Y406F were found to be structurally altered, the other cytochrome c oxidase variants were suitable for band assignment in the infrared. Reduced-minus-oxidized FTIR difference spectra of the mutant enzymes were used to identify the ring D propionate of heme a as a likely proton acceptor upon reduction of cytochromic oxidase. The ring D propionate of heme a(3) might undergo conformational changes or, less likely, act as a proton donor. PMID- 10684617 TI - pH-dependent unfolding of aspergillopepsin II studied by small-angle X-ray scattering. AB - Aspergillopepsin II (EC 3.4.23.6) secreted from the fungus Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase. It consists of two polypeptide chains (i.e., a heavy chain and a light chain), which are bound noncovalently to each other. The pH titration analysis using small-angle X-ray scattering (SAXS) as well as circular dichroism (CD) and gel filtration indicated that the enzyme was unfolded around a neutral pH with concomitant dissociation of the two chains. Detailed analyses showed that the midpoint pH values for the unfolding are not coincident with one another (pH 6.1 in circular dichroism and gel filtration, pH 6.4 in zero-angle intensity of SAXS, pH 6.8 in radius of gyration). The difference between these values suggested the existence of an intermediate state during the unfolding. Further analyses of the SAXS data showed that the heavy chain just after the dissociation still kept molecular compactness and that it gradually increased its dimensions as the pH was further raised. Noncoincidence of the two phenomena (i.e., chain dissociation and swelling) led to elucidation of a novel intermediate state during unfolding, which was confirmed by the subsequent singular value decomposition (SVD) analysis. PMID- 10684618 TI - Characterization of the second metal site on avian phosphoenolpyruvate carboxykinase. AB - Chicken liver phosphoenolpyruvate carboxykinase (PEPCK) requires two divalent cations for activity. One cation activates the enzyme through a direct interaction with the protein at site n(1). The second cation, at site n(2), acts in the cation-nucleotide complex that serves as a substrate. The Co(3+)(n(1)) PEPCK and Cr(3+)(n(1))-PEPCK complexes were used to examine the kinetic, mechanistic, and binding properties of the n(2) metal. EPR studies performed on the Co(3+)(n(1))-PEPCK-GTP complex yielded a stoichiometry of 1 mol of Mn(2+) bound per mole of Co(3+)(n(1))-PEPCK-GTP with a K(D) of 5 microM. PRR studies show a significant enhancement for the Co(3+)(n(1))-PEPCK-Mn(2+)(n(2))-GDP complex. A change in enhancement in the presence of PEP suggests that PEP interacts with the second metal ion. The distance between Mn(2+) at site n(2) on PEPCK and the cis and trans protons and the (31)P of PEP are 7.0, 7.5, and 4.8 A, respectively, as measured by high-resolution NMR. PRR studies of the Co(3+)(n(1)) PEPCK-Mn(2+)(n(2))-GTP and Co(3+)(n(1))-PEPCK-Mn(2+)(n(2))-GDP complexes as a function of frequency (omega(I)) were used to estimate the hydration number of the n(2) metal to be between 0.5 and 0.7. The metal-metal distance for the M(n(1))-PEPCK-M(n(2))-GTP complex is approximately 8.3 A, and the distance for the M(n(1))-PEPCK-M(n(2))-GDP complex is 9.2 A. The change in the metal-metal distance suggests a conformational change at the active site of PEPCK occurs during catalysis. The Co(3+)(n(1))-PEPCK complex was incubated with Co(2+), GTP, and H(2)O(2) to create a doubly labeled and inactive Co(3+)(n(1))-PEPCK Co(3+)(n(2))-GTP complex. The Co(3+)(n(1))-PEPCK-Co(3+)(n(2))-GTP complex was digested by LysC, and two cobalt-containing peptides were purified using RP-HPLC. Amino acid sequencing of the second cobalt-containing peptide points to the region of Tyr57-Lys76 of PEPCK. Asp66, Asp69, and Glu74 are all feasible ligands to the site n(2) metal. PMID- 10684619 TI - Solution 1H NMR study of the heme cavity and folding topology of the abbreviated chain 118-residue globin from the cyanobacterium Nostoc commune. AB - The globin from the cyanobacterium Nostoc commune, abbreviated GlbN, which appears to serve as a part of a terminal oxidase rather than as a respiratory pigment, displays relatively normal O2 binding properties, despite the highly abbreviated polypeptide chain, (118 residues) relative to more conventional globins [Thorsteinsson, M. V. , Bevan, D. R., Potts, M., Dou, Y., Eich, R. F., Hargrove, M. S., Gibson, Q. H., and Olson, J. S. (1999) Biochemistry 38, 2117 2126]. The nature of the heme cavity and the general folding topology of this cyanoglobin were investigated by solution 1H NMR to establish the extent to which, and the manner in which, this compact globin adheres to the standard globin fold. This represents by far the smallest globin subjected to structural analysis. The paramagnetic cyanomet derivative was selected because its characteristically large magnetic anisotropy imparts significant dipolar shifts which both improve resolution to greatly facilitate assignments and serve as indicators of the folding topology of the globin. Identification of the axial His 70 and highly conserved Phe 35 (CD1) determined the absolute orientation of the heme and proximal His. Sequential assignments of four helical and one loop segments, which exhibit dipolar contacts to the heme and among each other, confirm the presence of well-conserved F, G, and H helices and the FG corner. The majority of the abbreviation of the chain relative to the more conventional length globins is accommodated in the A-D helices, of which the last is completely missing. The distal residue which provides a H-bond to bound ligand is identified as Gln 43, but the expected helical position E7 could not be confirmed. His 46, placed at position E10, is found to adopt alternate orientations into, and out of, the heme cavity depending on protonation state, suggesting the presence of a Bohr effect at low pH. It is shown that the dipolar shifts exhibited by backbone protons for the assigned residues conform well to those observed for other cyanomet globins and further support a conserved Mb fold. Perturbed medium-range dipolar contacts and the pH-independent backbone proton lability of the F helix are interpreted in terms of a holoprotein which is less stable than a conventional length globin. PMID- 10684620 TI - Iso-mechanism of nitroalkane oxidase: 1. Inhibition studies and activation by imidazole. AB - The flavoprotein nitroalkane oxidase catalyzes the oxidation of primary and secondary nitroalkanes to aldehydes and ketones, respectively, transferring electrons to oxygen to form hydrogen peroxide. The steady-state kinetic mechanism of the active flavin adenine dinucleotide-(FAD-) containing form of the enzyme has been determined with nitroethane at pH 7 to be bi-ter ping-pong, with oxygen reacting with the free reduced enzyme after release of the aldehyde product. The V(max) value is 5.5 +/- 0.3 s(-)(1) and the K(m) values for nitroethane and oxygen are 3.3 +/- 0.6 and 0.023 +/- 0.007 mM, respectively. The free reduced enzyme forms a dead-end complex with nitroethane, with a K(ai) value of 30 +/- 6 mM. Acetaldehyde and butyraldehyde are noncompetitive inhibitors versus nitroethane due to formation of a dead-end complex between the oxidized enzyme and the product. Acetaldehyde is an uncompetitive inhibitor versus oxygen, indicating that an irreversible isomerization of the free reduced enzyme occurs before the reaction with oxygen. Addition of unprotonated imidazole results in a 5-fold increase in the V(max) value, while the V/K values for nitroethane and oxygen are unaffected. A 5-fold increase in the K(ai) value for nitroethane and a 6.5-fold increase in the K(ii) value for butyraldehyde are observed in the presence of imidazole. These results are consistent with the isomerization of the free reduced enzyme being about 80% rate-limiting for catalysis and with a model in which unprotonated imidazole accelerates the rate of isomerization. PMID- 10684621 TI - Mechanism of nitroalkane oxidase: 2. pH and kinetic isotope effects. AB - Nitroalkane oxidase catalyzes the oxidation of nitroalkanes to aldehydes or ketones with production of nitrite and hydrogen peroxide. pH and kinetic isotope effects with [1, 1-(2)H(2)]nitroethane have been used to study the mechanism of this enzyme. The V/K(ne) pH profile is bell-shaped. A group with a pK(a) value of about 7 must be unprotonated and one with a pK(a) value of 9.5 must be protonated for catalysis. The lower pK(a) value is seen also in the pK(is) profile for the competitive inhibitor valerate, indicating that nitroethane has no significant external commitments to catalysis. The (D)(V/K)(ne) value is pH-independent with a value of 7.5, whereas the (D)V(max) value increases from 1.4 at pH 8.2 to a limiting value of 7.4 below pH 5. The V(max) pH profile decreases at low and high pH, with pK(a) values of 6.6 and 9.5, respectively. Imidazole, which activates the enzyme, affects the V(max) but not the V/K(ne) pH profile. In the presence of imidazole at pH 7 the (D)V(max) value increases to a value close to the intrinsic value, consistent with cleavage of the carbon-hydrogen bond of the substrate being fully rate-limiting for catalysis in the presence of imidazole. PMID- 10684622 TI - pH-dependent changes in the in vitro ligand-binding properties and structure of human clusterin. AB - Clusterin is a glycoprotein which is locally overexpressed at sites of tissue damage or stress, leading to the proposal that it may be a cytoprotective protein. It has been shown that clusterin has chaperone-like activity, being able to protect proteins against precipitation under stress conditions. It has also been shown that local acidosis is common at sites of tissue damage or stress. We asked whether acidic pH induces structural changes in clusterin and enhances its ability to bind to other proteins. We found by affinity chromatography and ELISA that the binding of clusterin to glutathione-S-transferase, IgG, apolipoprotein A I, and complement protein C9 was enhanced at mildly acidic compared to physiological pH. Analytical ultracentrifugation and gel filtration studies revealed that clusterin exists in different polymerization states with monomer occurring preferentially at pH 5.5 and multimeric species at pH 7.5. Although circular dichroism showed little difference in the alpha-helical and beta-sheet contents of clusterin at pH 5 compared to pH 7.5, evidence for pH-dependent structural changes in clusterin was obtained from fluorescence experiments. pH titrations showed reversible changes in the fluorescence of tryptophan residues in clusterin. There was a reversible 2-fold increase in the fluorescence of the extrinsic probe 4, 4'-bis(1-anilinonaphthalene-8-sulfonate) bound to clusterin at pH 5. 5 compared to pH 7.5. There was also a 3.5-fold increase in fluorescence resonance energy transfer from tryptophan residues in clusterin to 4,4'-bis(1 anilinonaphthalene-8-sulfonate) at pH 5.5 compared to pH 7.5. These data suggest that pH-induced changes in the structure of clusterin are responsible for its enhanced ability to bind protein ligands at mildly acidic pH. PMID- 10684623 TI - Mutation of R116C results in highly oligomerized alpha A-crystallin with modified structure and defective chaperone-like function. AB - An autosomal dominant congenital cataract in human is associated with mutation of Arg-116 to Cys (R116C) in alpha A-crystallin. To investigate the molecular basis of cataract formation, rat alpha A-crystallin cDNA was cloned into pET-23d(+), and the site-directed mutants S142C (similar to wild-type human alpha A) and R116C/S142C or R116C (similar to human R116C variant) were generated. These were expressed in E. coli and the recombinant alpha A-crystallins purified by Sephacryl size-exclusion chromatography. The chaperone-like function of mutant R116C determined at 37 degrees C with insulin and alcohol dehydrogenase as target proteins was about 40% lower than those of wild-type and mutant S142C. Based on size-exclusion chromatography data, the oligomeric size of the R116C mutant was about 2000 kDa at 25 degrees C, 1400 kDa at 37 degrees C, and 900 kDa at 45 degrees C. In comparison, alpha A-wild-type and alpha A-S142C ranged from 477 to 581 kDa. Heat stability studies corroborated the effect of temperature on the dynamic quaternary structure of the R116C mutant. Circular dichroism spectra showed secondary and tertiary structural changes, and ANS fluorescence spectra showed loss of surface hydrophobicity in the R116C mutant. These findings suggest that the molecular basis for the congenital cataract with the alpha A-R116C mutation is due to the generation of a highly oligomerized alpha A-crystallin having a modified structure and decreased chaperone-like function. PMID- 10684624 TI - Human immunodeficiency virus type 1 reverse transcriptase dimer destabilization by 1-[Spiro[4"-amino-2",2" -dioxo-1",2" -oxathiole-5",3'-[2', 5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl]]]-3-ethylthy mine. AB - The nonnucleoside inhibitor binding pocket is a well-defined region in the p66 palm domain of the human immunodeficiency virus type-1 reverse transcriptase (HIV 1 RT). This binding pocket opens toward the interface of the p66/p51 heterodimer and we have investigated whether ligand binding at or near this site induces structural changes that have an impact on the dimeric structure of HIV-1 RT. 1 [2',5'-bis-O-(tert-butyldimethylsilyl]-3'-spiro-5' '-(4' '-amino-1' ',2' ' oxathiole-2' ',2' '-dioxide)-3-ethylthymine (TSAOe(3)T) was found to destabilize the subunit interactions of both the p66/p51 heterodimer and p66/p66 homodimer enzymes. The Gibbs free energy of dimer dissociation (DeltaG(D)(H)2(O)) is decreased with increasing concentrations of TSAOe(3)T, resulting in a loss in dimer stability of 4.0 and 3.2 kcal/mol for the p66/p51 and p66/p66 HIV-1 RT enzymes, respectively. This loss of energy is not sufficient to induce the dissociation of the subunits in the absence of denaturant. This destabilizing effect seems to be unique for TSAOe(3)T, since neither the tight-binding inhibitor UC781 nor nevirapine showed any effects on the stability of HIV-1 RT dimers. TSAOe(3)T was unable to destabilize the subunit interactions of the E138K mutant enzyme, which exhibits significant resistance to TSAOe(3)T inhibition. Molecular modeling of TSAOm(3)T into the nonnucleoside inhibitor binding pocket of wild-type RT suggests that it makes significant interactions with the p51 subunit of the enzyme, a feature that has not been observed with other types of nonnucleoside inhibitors. The observed destabilization of the dimeric HIV-1 RT may result from structural/conformational perturbations at the reverse transcriptase subunit interface. PMID- 10684625 TI - Potentiation and inhibition of bFGF binding by heparin: a model for regulation of cellular response. AB - Basic fibroblast growth factor (bFGF) binds to cell surface tyrosine kinase receptor proteins and to heparan sulfate proteoglycans. The interaction of bFGF with heparan sulfate on the cell surface has been demonstrated to impact receptor binding and biological activity. bFGF receptor binding affinity is reduced on cells that do not express heparan sulfate. The addition of soluble heparin or heparan sulfate has been demonstrated to rescue the bFGF receptor binding affinity on heparan sulfate deficient cells yet has also been shown to inhibit binding under some conditions. While the chemical requirements of the heparin bFGF-receptor interactions have been studied in detail, the possibility that heparin enhances bFGF binding in part by physically associating with the cell surface has not been fully evaluated. In the study presented here, we have investigated the possibility that heparin binding to the cell surface might play a role in modulating bFGF receptor binding and activity. Balb/c3T3 cells were treated with various concentrations of sodium chlorate, so as to express a range of endogenous heparan sulfate sites, and [(125)I]bFGF binding was assessed in the presence of a range of heparin concentrations. Low concentrations of heparin (0.1 30 nM) enhanced bFGF receptor binding to an extent that was inversely proportional to the amount of endogenous heparan sulfate sites present. At high concentrations (10 microM), heparin inhibited bFGF receptor binding in cells under all conditions. The ability of heparin to stimulate and inhibit bFGF receptor binding correlated with altered bFGF-stimulated tyrosine kinase activity and cell proliferation. Under control and chlorate-treated conditions, [(125) I]heparin was observed to bind with a high affinity to a large number of binding sites on the cells (K(d) = 57 and 50 nM with 3.5 x 10(6) and 3.6 x 10(6) sites/cell for control and chlorate-treated cells, respectively). A mathematical model of this process revealed that the dual functions of heparin in bFGF binding were accurately represented by heparin cell binding-mediated stimulation and soluble heparin-mediated inhibition of bFGF receptor binding. PMID- 10684626 TI - Investigations of the myoglobin cavity mutant H93G with unnatural imidazole proximal ligands as a modular peroxide O-O bond cleavage model system. AB - A general inability to elucidate extensive variations in the electronic characteristics of proximal heme iron ligands in heme proteins has hampered efforts to obtain a clear understanding of the role of the proximal heme iron ligand in the activation of oxygen and peroxide. The disadvantage of the frequently applied site-directed mutagenesis technique is that it is limited by the range of natural ligands available within the genetic code. The myoglobin cavity mutant H93G [Barrick, D. (1994) Biochemistry 33, 6546-6554] has its proximal histidine ligand replaced with glycine, a mutation which leaves an open cavity capable of accommodating a variety of unnatural potential proximal ligands. We have carried out investigations of the effect of changing the electron donor characteristics of a variety of substituted imidazole proximal ligands on the rate of formation of myoglobin compound II and identified a correlation between the substituted imidazole N-3 pK(a) (which provides a measure of the electron donor ability of N-3) and the apparent rate of formation of compound II. A similar rate dependence correlation is not observed upon binding of azide. This finding indicates that O-O bond cleavage and not the preceding peroxide binding step is being influenced by the electron donor characteristics of the substituted imidazole ligands. The proximal ligand effects are clearly visible, but their overall magnitude is quite low (1.7-fold increase in the O-O bond cleavage rate per pK(a) unit). This appears to provide support for recent commentaries which concluded that the partial ionization of the proximal histidine ligand in typical heme peroxidases may not be enough of an influence to provide a mechanistically critical push effect [Poulos, T. L. (1996) JBIC, J. Biol. Inorg. Chem. 1, 356-359]. Further attempts were made to define the mechanism of the influence of N-3 pK(a) on O-O bond cleavage by using peracetic acid and cumene hydroperoxide as mechanistic probes. The observation of heme destruction in these reactions indicates that displacement of the proximal imidazole ligands by peracetic acid or cumene hydroperoxide has occurred. A combination mutation (H64D/H93G) was prepared with the objective of observing compound I of H64D/H93G with substituted imidazoles as proximal ligands upon reaction with H(2)O(2). This double mutant was found to simultaneously bind imidazole to both axial positions, an arrangement which prevents a reaction with H(2)O(2). PMID- 10684627 TI - Hydrogen exchange at the core of Escherichia coli alkaline phosphatase studied by room-temperature tryptophan phosphorescence. AB - The room-temperature tryptophan (Trp) phosphorescence lifetime is sensitive to details of the local environment and has been shown to increase significantly in some proteins following H-D exchange. Careful analysis of the phosphorescence lifetime distribution of Trp 109 in Escherichia coli alkaline phosphatase (AP) in solution as a function of time during the H-D exchange shows that this process corresponds to a two-state reaction resulting from the deuteration of a single, specific hydrogen in the core of the protein. The absence of a pH dependence of the exchange rate suggests that the exchange is not an EX2 process, and therefore, a certain degree of unfolding is required for exchange to occur. This discovery opens up the use of phosphorescence-detected hydrogen exchange as a sensitive tool for monitoring the local susceptibility and activation energy for exchange in proteins having a phosphorescent Trp and, for example, for studying the effects of local mutations upon that susceptibility. PMID- 10684628 TI - Effect of loop sequence and size on DNA aptamer stability. AB - The thrombin aptamer is a 15-mer oligodeoxyribonucleotide that folds into a unimolecular quadruplex consisting of a stack of two guanine quartets connected by two external loops and one central loop and possesses a high affinity for thrombin. We have undertaken a systematic examination, in KCl, of the thermodynamic stability of thrombin aptamer analogues containing sequence modifications in one or more of the loops, as well as in the number of quartets. UV melting studies have been carried out to obtain the relevant thermodynamic parameters for these aptamers. van't Hoff analysis of these data, with a two state model for unimolecular denaturation, gave excellent fits to the experimental observations. Thermodynamic analysis indicates that the central loop sequence in the parent aptamer is optimal for stability. Modifications in this or other loops can effect either DeltaH degrees, DeltaS degrees, or both. Addition of a single G at the 5'-end decreases stability while addition of a G at the 3' end increases stability. Differential scanning calorimetry experiments on the thrombin aptamer reveal that a heat capacity change, not detected by UV measurements, accompanies the unfolding of the aptamer. PMID- 10684629 TI - Binding of fatty acids and peroxisome proliferators to orthologous fatty acid binding proteins from human, murine, and bovine liver. AB - Liver-type fatty acid binding protein (L-FABP) has been proposed to be involved in the transport of fatty acids and peroxisome proliferators from the cytosol into the nucleus for interaction with the peroxisome proliferator-activated receptors (PPARs). On the basis of this premise, we investigated by isothermal titration calorimetry the binding of myristic, stearic, oleic, and docosahexaenoic acids to three orthologous L-FABPs and compared these results to those obtained for several xenobiotics [Wy14,643, bezafibrate, 5,8,11,14 eicosatetraynoic acid (ETYA), and BRL48,482] known for their peroxisome proliferating activity in rodents. Recombinant human, murine, and bovine L-FABPs were analyzed and the thermodynamic data were obtained. Our studies showed that fatty acids bound with a stoichiometry of 2:1, fatty acid to protein, with dissociation constants for the first binding site in the nanomolar range. With dissociation constants above 1 microM the drug peroxisome proliferators showed weaker binding, with the exception of arachidonate analogue ETYA, which bound with a similar affinity as the natural fatty acid. Some of the thermodynamic data obtained for fatty acid binding could be explained by differences in protein structure. Moreover, our results revealed that binding affinities were not determined by ligand solubility in the aqueous phase. PMID- 10684630 TI - Insulin-like signaling in yeast: modulation of protein phosphatase 2A, protein kinase A, cAMP-specific phosphodiesterase, and glycosyl-phosphatidylinositol specific phospholipase C activities. AB - Previously, we have described significant effects of human insulin on glucose metabolism in the yeast Saccharomyces cerevisiae under conditions of growth limitation. These regulations apparently rely on a transmembrane receptor capable of binding human insulin and responding by tyrosine/serine phosphorylation of a specific set of polypeptides [Muller, G., Rouveyre, N., Crecelius, A., and Bandlow, W. (1998) Biochemistry 37, 8683-8695; Muller, G., Rouveyre, N., Upshon, C., Gross, E., and Bandlow, W. (1998) Biochemistry 37, 8696-8704; Muller, G., Rouveyre, N., Upshon, C., and Bandlow, W. (1998) Biochemistry 37, 8705-8713]. To characterize the molecular link between the initial steps in insulin-like signaling in yeast and the changes in the activities of glycogen synthase and glycogen phosphorylase, we examined here the effects of human insulin on a set of key regulatory enzymes of glycogen metabolism, protein phosphatase 2A (PP2A), cAMP-specific phosphodiesterase (cAMP-PDE), and protein kinase A (PKA). PP2A was activated about 2-fold by insulin in spheroplasts and in intact cells, whereas the fraction of active PKA was significantly reduced in a cAMP-independent manner as well as through a subsequent up to 3-fold increase in particulate cAMP-PDE activity accompanied by a 50% decrease in cytosolic cAMP levels. In addition, glycosyl-phosphatidylinositol-specific phospholipase C (GPI-PLC), which in isolated rat adipocytes is activated by insulin, was stimulated to up to 5-fold by glucose and 10-fold by glucose plus insulin in both yeast spheroplasts and intact cells leading to a concentration-dependent leftward shift of the glucose response curve for activation of the GPI-PLC. GPI-PLC was most pronouncedly stimulated by authentic human insulin compared to various insulin analogues and insulin-like growth factor I. In addition to lipolytic cleavage by GPI-PLC, the GPI anchor of the cAMP-binding ectoprotein, Gce1p, was secondarily processed by a rapid proteolytic event. As the GPI-PLC reaction is rate limiting, the efficiency of the two-step anchor cleavage was significantly increased when insulin was present together with glucose as compared to glucose alone. The insulin concentrations effective in modulating PP2A, PKA, cAMP-PDE, and GPI-PLC activities correlate well with those required for half-saturation of the specific binding sites as well as for stimulation of protein phosphorylation and glycogen accumulation. The data suggest that mammalian insulin-sensitive cells and yeast share (part of) the key regulatory mechanism (consisting of PP2A, PKA, cAMP-PDE, and GPI-PLC) involved in the transduction of the insulin signal from the respective receptor systems to glycogen synthase and phosphorylase. PMID- 10684631 TI - Excitation dynamics and heterogeneity of energy equilibration in the core antenna of photosystem I from the cyanobacterium Synechocystis sp. PCC 6803. AB - Energy equilibration in the photosystem I core antenna from the cyanobacterium Synechocystis sp. PCC 6803 was studied using femtosecond transient absorption spectroscopy at 298 K. The photosystem I core particles were excited at 660, 693, and 710 nm with 150 fs spectrally narrow laser pulses (fwhm = 5 nm). Global analysis revealed three kinetic processes in the core antenna with lifetimes of 250-500 fs, 1.5-2.5 ps, and 20-30 ps. The first two components represent strongly excitation wavelength-dependent energy equilibration processes while the 20-30 ps phase reflects the trapping of energy by the reaction center. Excitation into the blue and red edge of the absorption band induces downhill and uphill energy flows, respectively, between different chlorophyll a spectral forms of the core. Excitation at 660 nm induces a 500 fs downhill equilibration process within the bulk of antenna while the selective excitation of long-wavelength-absorbing chlorophylls at 710 nm results in a 380 fs uphill energy transfer to the chlorophylls absorbing around 695-700 nm, presumably reaction center pigments. The 1.5-2.5 ps phases of downhill and uphill energy transfer are largely equivalent but opposite in direction, indicating energy equilibration between bulk antenna chlorophylls at 685 nm and spectral forms absorbing below 700 nm. Transient absorption spectra with excitation at 693 nm exhibit spectral evolution within approximately 2 ps of uphill energy transfer to major spectral forms at 680 nm and downhill energy transfer to red pigments at 705 nm. The 20-30 ps trapping component and P(700) photooxidation spectra derived from data on the 100 ps scale are largely excitation wavelength independent. An additional decay component of red pigments at 710 nm can be induced either by selective excitation of red pigments or by decreasing the temperature to 264 K. This component may represent one of the phases of energy transfer from inhomogeneously broadened red pigments to P(700). The data are discussed based on the available structural model of the photosystem I reaction center and its core antenna. PMID- 10684632 TI - Structural consequences of b- to c-type heme conversion in oxidized Escherichia coli cytochrome b562. AB - An NMR characterization of the 98Arg --> Cys variant of iron (III)-containing cytochrome b562 from Escherichia coli has been performed and the solution structure obtained. This variant has a covalent bond between the heme and Cys 98, thus mimicking the heme binding in cytochrome c. The R98C cytochrome is shown to have a significantly increased stability, compared to that of wild type, toward thermal and chemical denaturation. In water at 20 degrees C it is 5.60 kJ mol-1 more stable than the WT protein, measured by equilibrium guanidine hydrochloride denaturation. The structure has been obtained through two-dimensional total correlation spectroscopy (TOCSY) and nuclear Overhauser effect spectroscopy (NOESY) experiments and through three-dimensional NOESY-15N heteronuclear multiple quantum coherence (HMQC). By these methods, 85% of protons and 100% of backbone nitrogens were assigned. 2145 meaningful nuclear Overhauser effects (NOEs) (20 NOEs per residue), 45 backbone 3J values, and 397 pseudocontact shifts were used to obtain a family of 35 members, which were then energy-minimized. The root-mean-square deviation (RMSD) with respect to the average structure is 0.50 +/- 0.07 for the backbone and 1.01 +/- 0.08 for the heavy atoms. The magnetic anisotropy resulting from analysis of the pseudocontact shifts indicates an anisotropy that is an intermediate between that of the wild-type, which is the smallest, and cytochrome c. The g values confirm a higher anisotropy of the variant with respect to the wild-type protein. The chirality of the heme 2 alpha carbon is the same as that in all naturally occurring cytochromes c. The overall secondary structure and tertiary structure are very similar to the wild type. The removal of Arg 98 causes a change in the pH-dependent properties. The pKa, proposed to be due to deprotonation of the coordinated histidine, is 1.5 units higher than in the wild type, consistent with the lack of the positive charge of Arg 98 close to the ionizable group. This is further support for the coordinated histidine being the titratable group with an alkaline pKa in the wild-type protein. The pattern of the shifts of the heme methyl groups is different than in the wild-type protein, presumably due to alteration of the electronic structure by the presence of the covalent bond between the protein and the heme. The difference in stability between the variant and wild-type protein is discussed in terms of the structural information. PMID- 10684633 TI - Cloning of amadoriase I isoenzyme from Aspergillus sp.: evidence of FAD covalently linked to Cys342. AB - Amadoriases are a novel class of FAD enzymes which catalyze the oxidative deglycation of glycated amino acids to yield corresponding amino acids, glucosone, and H(2)O(2). We previously reported the purification and characterization of two amadoriase isoenzymes from Aspergillus fumigatus and the molecular cloning of amadoriase II. To identify the primary structure of amadoriase I, we prepared a cDNA library from Aspergillus fumigatus and isolated a clone using a probe amplified by polymerase chain reaction with primers designed according to the partial amino acid sequences from peptide mapping. The primary structure of the enzyme deduced from the nucleotide sequence comprises 445 amino acid residues. The enzyme contains 1 mol of FAD as a cofactor, which is covalently linked to Cys342, as determined by mutagenesis analysis, matrix assisted laser desorption/ionization time-of-flight mass spectrometry, and electrospray ionization-collisional-activated dissociation tandem mass spectrometry. Sequence alignment studies show that amadoriase I has 22% homology with monomeric sarcosine oxidase in which FAD is also linked to a homologous Cys residue. Amadoriases are of potential importance as tools for uncoupling hyperglycemia and glycation reactions that are thought to play a role in diabetic complications. PMID- 10684634 TI - Catalytic mechanism of a C-C hydrolase enzyme: evidence for a gem-diol intermediate, not an acyl enzyme. AB - 2-Hydroxy-6-keto-nona-2,4-diene 1,9-dioic acid 5,6-hydrolase (MhpC) from Escherichia coli catalyses the hydrolytic cleavage of the extradiol ring fission product on the phenylpropionate catabolic pathway and is a member of the alpha/beta hydrolase family. The catalytic mechanism of this enzyme has previously been shown to proceed via initial ketonization of the dienol substrate (Henderson, I. M. J., and Bugg, T. D. H. (1997) Biochemistry 36, 12252-12258), followed by stereospecific fragmentation. Despite the implication of an active site serine residue in the alpha/beta hydrolase family, attempts to verify a putative acyl enzyme intermediate by radiochemical trapping methods using a (14)C labeled substrate yielded a stoichiometry of <1% covalent intermediate, which could be accounted for by nonenzymatic processes. In contrast, incorporation of 5 6% of two atoms of (18)O from H(2)(18)O into succinic acid was observed using the natural substrate, consistent with the reversible formation of a gem-diol intermediate. Furthermore, time-dependent incorporation of (18)O from H(2)(18)O into the carbonyl group of a nonhydrolysable analogue 4-keto-nona-1,9-dioic acid was observed in the presence of MhpC, consistent with enzyme-catalyzed attack of water at the ketone carbonyl. These results favor a catalytic mechanism involving base-catalyzed attack of water, rather than nucleophilic attack of an active site serine. The implication of this work is that the putative active site serine in this enzyme may have an alternative function, for example, as a base. PMID- 10684635 TI - Superinduction of cyclooxygenase-2 by NO(*) and agonist challenge involves transcriptional regulation mediated by AP-1 activation. AB - Superinduction of cyclooxygenase-2, in murine RAW 264.7 macrophages as well as human pulmonary type II A549 epithelial cells, is achieved by the simultaneous addition of agonists such as lipopolysaccharide or interleukin-1beta and the NO(*) donor S-nitrosoglutathione. NO(*)-evoked superinduction of cyclooxygenase-2 in the presence of agonists was dose-dependent and required transcriptional as well as translational regulation. We sought to further analyze NO(*)-elicited superinduction at the level of the transcription factor NF-kappaB that is obligatory for cyclooxygenase-2 expression. NO(*)-mediated NF-kappaB activation was restricted to low concentrations of S-nitrosoglutathione (50-200 microM), while a higher dose of S-nitrosoglutathione (1 mM) was ineffective. Not observing a correlation between NF-kappaB activation and cyclooxygenase-2 expression under NO(*)-delivery stimulated our interest in analyzing AP-1. NO(*) efficiently activated AP-1 at all concentrations tested. The involvement of AP-1 in promoting cyclooxygenase-2 superinduction was established in cells transfected with the dominant-negative c-Jun mutant, TAM-67. Enhanced expression of cyclooxygenase-2 by lipopolysaccharide/S-nitrosoglutathione-treatment was attenuated in TAM-67 transfectants, while the response to lipopolysaccharide alone remained unaffected. We conclude that AP-1 activation exclusively conveys the NO(*) signal that is required for superinduction of cyclooxygenase-2. Superinduction of cyclooxygenase-2 is restricted to a situation where both, NF-kappaB and AP-1 are activated. Under inflammatory conditions this might be achieved by the costimulatory signals provided by agonist challenge and NO(*). PMID- 10684636 TI - Characterization of the binding interface between the E-domain of staphylococcal protein A and an antibody Fv-fragment PMID- 10684637 TI - Abstracts, division of biological chemistry, 219th national meeting of the american chemical society, march 26-30, 2000 PMID- 10684638 TI - Structure of sterol carrier protein 2 at 1.8 A resolution reveals a hydrophobic tunnel suitable for lipid binding. AB - Sterol carrier protein 2, also known as nonspecific lipid transfer protein is a ubiquitous, small, basic protein of 13 kDa found in animals. Its primary structure is highly conserved between different species, and it has been implicated in the intracellular transport of lipids and in a wide range of other in vitro functions related to sterol and fatty acid metabolism. Sterol carrier protein 2 deficiency in mice leads to elevated concentrations of phytanic acid in the serum and causes hepatocarcinogenesis. However, its actual physiological role is still unknown. Although sterol carrier protein 2 has been studied extensively in the past 20 years, very little is known concerning its three-dimensional structure. The crystal structure of rabbit sterol carrier protein 2, determined at 1.8 A resolution with the MIRAS method, shows a unique alpha/beta-fold. The core of this protein forms a five-stranded antiparallel beta-sheet flanked by five helices. A C-terminal segment (residues 114-123), together with part of the beta-sheet and four alpha-helices, form a hydrophobic tunnel providing the environment for apolar ligands such as fatty acids and fatty acyl-coenzyme As. Structurally well-characterized nonspecific lipid transfer proteins from plants have hydrophobic tunnel-like cavities, which were identified as the binding site for fatty acids and related apolar ligands. Despite the fact that plant nonspecific lipid transfer proteins are smaller proteins than sterol carrier protein 2, show no sequence homology to sterol carrier protein 2, and are structurally unrelated, the cavities of these two classes of proteins are very similar with respect to size, shape, and hydrophobicity, suggesting a common functional role. PMID- 10684639 TI - Crystal structures of a low-molecular weight protein tyrosine phosphatase from Saccharomyces cerevisiae and its complex with the substrate p-nitrophenyl phosphate. AB - Low-molecular weight protein tyrosine phosphatases are virtually ubiquitous, which implies that they have important cellular functions. We present here the 2.2 A resolution X-ray crystallographic structure of wild-type LTP1, a low molecular weight protein tyrosine phosphatase from Saccharomyces cerevisiae. We also present the structure of an inactive mutant substrate complex of LTP1 with p nitrophenyl phosphate (pNPP) at a resolution of 1.7 A. The crystal structures of the wild-type protein and of the inactive mutant both have two molecules per asymmetric unit. The wild-type protein crystal was grown in HEPES buffer, a sulfonate anion that resembles the phosphate substrate, and a HEPES molecule was found with nearly full occupancy in the active site. Although the fold of LTP1 resembles that of its bovine counterpart BPTP, there are significant changes around the active site that explain differences in their kinetic behavior. In the crystal of the inactive mutant of LTP1, one molecule has a pNPP in the active site, while the other has a phosphate ion. The aromatic residues lining the walls of the active site cavity exhibit large relative movements between the two molecules. The phosphate groups present in the structures of the mutant protein bind more deeply in the active site (that is, closer to the position of nucleophilic cysteine side chain) than does the sulfonate group of the HEPES molecule in the wild-type structure. This further confirms the important role of the phosphate-binding loop in stabilizing the deep binding position of the phosphate group, thus helping to bring the phosphate close to the thiolate anion of nucleophilic cysteine, and facilitating the formation of the phosphoenzyme intermediate. PMID- 10684640 TI - Crystal structure of flavocetin-A, a platelet glycoprotein Ib-binding protein, reveals a novel cyclic tetramer of C-type lectin-like heterodimers. AB - Snake venom contains a number of the hemostatically active C-type lectin-like proteins, which affect the interaction between von Willebrand factor (vWF) and the platelet glycoprotein (GP) Ib or platelet receptor to inhibit/induce platelet activation. Flavocetin-A (FL-A) is a high-molecular mass C-type lectin-like protein (149 kDa) isolated from the habu snake venom. FL-A binds with high affinity to the platelet GP Ibalpha-subunit and functions as a strong inhibitor of vWF-dependent platelet aggregation. We have determined the X-ray crystal structure of FL-A and refined to 2.5 A resolution. This is a first elucidation of a three-dimensional structure of the platelet GP Ib-binding protein. The overall structure reveals that the molecule is a novel cyclic tetramer (alphabeta)(4) made up of four alphabeta-heterodimers related by a crystallographic 4-fold symmetry. The tetramerization is mediated by an interchain disulfide bridge between cysteine residues at the C-terminus of the alpha-subunit and at the N terminus of the beta-subunit in the neighboring alphabeta-heterodimer. The high affinity of FL-A for the platelet GP Ib alpha-subunit could be explained by a cooperative-binding action through the multiple binding sites of the tetramer. PMID- 10684641 TI - Crystallographic studies of the interactions of Escherichia coli lytic transglycosylase Slt35 with peptidoglycan. AB - Lytic transglycosylases catalyze the cleavage of the beta-1, 4-glycosidic bond between N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc) in peptidoglycan with concomitant formation of a 1,6-anhydro bond in the MurNAc residue. To understand the reaction mechanism of Escherichia coli lytic transglycosylase Slt35, three crystal structures have been determined of Slt35 in complex with two different peptidoglycan fragments and with the lytic transglycosylase inhibitor bulgecin A. The complexes define four sugar-binding subsites (-2, -1, +1, and +2) and two peptide-binding sites in a large cleft close to Glu162. The Glu162 side chain is between the -1 and +1 sugar-binding sites, in agreement with a function as catalytic acid/base. The complexes suggest additional contributions to catalysis from Ser216 and Asn339, residues which are conserved among the MltB/Slt35 lytic transglycosylases. PMID- 10684642 TI - Ligand preference inferred from the structure of neutrophil gelatinase associated lipocalin. AB - Neutrophil gelatinase associated lipocalin (NGAL), a constituent of neutrophil granules, is a member of the lipocalin family of binding proteins. NGAL can also be highly induced in epithelial cells in both inflammatory and neoplastic colorectal disease. NGAL is proposed to mediate inflammatory responses by sequestering neutrophil chemoattractants, particularly N-formylated tripeptides and possibly leukotriene B(4) and platelet activating factor. The crystal structures of NGAL display a typical lipocalin fold, albeit with an unusually large and atypically polar binding site, or calyx. The fold of NGAL is most similar to the epididymal retinoic acid-binding protein, another lipocalin, though the overall architecture of the calyces are very different. The crystal structures also reveal either sulfate ions or an adventitiously copurified fatty acid bound in the binding site. Neither ligand is displaced by added N-formylated tripeptides. The size, shape, and character of the NGAL calyx, as well as the low relative affinity for N-formylated tripeptides, suggest that neither the copurified fatty acid nor any of the proposed ligands are likely to be the preferred ligand of this protein. Comparisons between the crystal structures and the recently reported solution structure of NGAL reveal significant differences, in terms of both the details of the structure and the overall flexibility of the fold. PMID- 10684643 TI - The single mutation Phe173 --> Ala induces a molten globule-like state in murine interleukin-6. AB - A series of three aromatic to alanine mutants of recombinant murine interleukin-6 lacking the 22 N-terminal residues (DeltaN22mIL-6) were constructed to investigate the role of these residues in the structure and function of mIL-6. While Y78A and Y97A have activities similar to that of DeltaN22mIL-6, F173A lacks biological activity. F173A retains high levels of secondary structure, as determined by far-UV circular dichroism (CD), but has substantially reduced levels of tertiary structure, as determined by near-UV CD and (1)H NMR spectroscopy. F173A also binds the hydrophobic dye 1-anilino-8 naphthalenesulfonic acid (ANS) over a range of pH values and exhibits noncooperative equilibrium unfolding (as judged by the noncoincidence of monophasic unfolding transitions monitored by far-UV CD and lambda(max), with midpoints of unfolding at 2.6 +/- 0. 1 and 3.5 +/- 0.3 M urea, respectively, and the lack of an observable thermal unfolding transition). These are all properties of molten globule states, suggesting that the loss of activity of F173A results from the disruption of the fine structure of the protein, rather than from the loss of a side chain that is important for ligand-receptor interactions. Surprisingly, under some conditions, this loosened conformation is no more susceptible to proteolytic attack than the parent protein. By analogy with human IL-6, Phe173 in DeltaN22mIL-6 makes multiple interhelical interactions, the removal of which appear to be sufficient to induce a molten globule-like conformation. PMID- 10684644 TI - Identification of functionally important amino acid residues within the C2-domain of human factor V using alanine-scanning mutagenesis. AB - We have previously determined that the C2-domain of human factor V (residues 2037 2196) is required for expression of cofactor activity and binding to phosphatidylserine (PS)-containing membranes. Naturally occurring factor V inhibitors and a monoclonal antibody (HV-1) recognized epitopes in the amino terminus of the C2-domain (residues 2037-2087) and blocked PS binding. We have now investigated the function of individual amino acids within the C2-domain using charge to alanine mutagenesis. Charged residues located within the C2 domain were changed to alanine in clusters of 1-3 mutations per construct. In addition, mutants W2063A, W2064A, (W2063, W2064)A, and L2116A were constructed as well. The resultant 30 mutants were expressed in COS cells using a B-domain deleted factor V construct (rHFV des B). All mutants were expressed efficiently based on the polyclonal antibody ELISA. The charged residues, Arg(2074), Asp(2098), Arg(2171), Arg(2174), and Glu(2189) are required for maintaining the structural integrity of the C2-domain of factor V. Four of these residues (Arg(2074), Asp(2098), Arg(2171), and Arg(2174)) correspond to positions in the factor VIII C-type domains that have been identified as point mutations in patients with hemophilia A. The epitope for the inhibitory monoclonal antibody HV 1 has been localized to Lys(2060) through Glu(2069) in the factor V C2-domain. The epitope for the inhibitory monoclonal antibody 6A5 is composed of amino acids His(2128) through Lys(2137). The PS-binding site in the factor V C2-domain includes amino acid residues Trp(2063) and Trp(2064). This site overlaps with the epitope for monoclonal antibody HV-1. These factor V C2-domain mutants should provide valuable tools for further defining the molecular interactions responsible for factor V binding to phospholipid membranes. PMID- 10684645 TI - Substrate specificity of human methylpurine DNA N-glycosylase. AB - The activity of human methylpurine DNA N-glycosylase (hMPG) for major substrates was directly compared using two types of substrates, i.e., natural DNA and synthetic oligonucleotides. By the use of ARP assay detecting abasic sites in DNA, we first investigated the activity on the natural DNA substrates containing methylpurines, ethenopurines, or hypoxanthine (Hx) prepared by the conventional methods. After the treatment with hMPG, the amount of AP sites in methylated DNA was much higher than that in DNA containing ethenopurines or Hx. The oligodeoxynucleotide having a single 7-methylguanine (7-mG) was newly synthesized in addition to 1, N(6)-ethenoadenine (epsilonA)-, Hx-, and 8-oxoguanine containing oligonucleotides. 7-mG was effectively excised by hMPG, though it might be less toxic than the other methylated bases with respect to mutagenesis and cell killing. The kinetic study demonstrated that k(cat)/K(m) ratios of the enzyme for epsilonA, Hx, and 7-mG were 2.5 x 10(-3), 1.4 x 10(-3), and 4 x 10(-4) min(-1) nM(-1), respectively. The oligonucleotides containing epsilonA effectively competed against 7-mG, while Hx substrates showed unexpectedly low competition. Concerning the effect of the base opposite damage, hMPG much preferred Hx.T to other Hx pairs, and epsilonA.C and epsilonA.A pairs were better substrates than epsilonA.T. PMID- 10684646 TI - p53 regulates caveolin gene transcription, cell cholesterol, and growth by a novel mechanism. AB - Transcription of the human caveolin gene, directed by a TATA-less promoter, is downregulated in actively dividing cells during S-phase, together with free cholesterol (FC) efflux. It is upregulated by medium low density lipoprotein FC levels in quiescent cells. In this study, a common mechanism has been identified to coordinate the growth- and FC-dependent expression of caveolin. In human skin fibroblasts, transcription factors E2F/DP-1 and Sp1 bound to adjacent consensus sites at -151 to -138 bp of the caveolin promoter DNA sequence in a complex stabilized by tumor suppressor protein p53. Wild-type p53 also bound directly to DNA to a caveolin promoter sequence containing two consensus half-sites (-292 to 283 bp and -273 to -264 bp) for this transcription factor. SREBP-1, previously identified as a transcriptional regulator of caveolin expression in response to FC, mediated its effect via the same E2F/Sp1 site. Overexpression of E2F or p53 increased E2F binding to the -148 to -141 bp site, increased FC efflux, and inhibited cell division. The mutant protein p53(143V-->A) was inactive. Okadaic acid, previously shown to inhibit growth, FC efflux, and caveolin expression, inhibited E2F/Sp1 binding, while higher concentrations of extracellular FC increased it. The present findings provide a molecular link between the cell cycle and FC homeostatic effects of caveolin. These results also describe a novel mechanism of action for p53 in a TATA-less gene promoter and provide further evidence for a significant regulatory role for FC in cell cycle progression. PMID- 10684647 TI - ATP-dependent dissociation of non-disulfide-linked aggregates of coagulation factor VIII is a rate-limiting step for secretion. AB - Deficiency in coagulation factor VIII leads to the bleeding disorder hemophilia A. Previous studies demonstrated that factor VIII secretion is limited due to an ATP-requiring step early in the secretory pathway. In this report, we identified that this ATP-dependent rate-limiting step involves the dissociation of non disulfide-linked aggregates within the endoplasmic reticulum (ER). In contrast to the numerous examples of interchain disulfide-linked aggregates, factor VIII is the first protein characterized to form non-disulfide-linked high molecular weight aggregates within the ER. Approximately a third of newly synthesized factor VIII was detected in high molecular weight aggregates. These aggregates disappeared over time as functional factor VIII appeared in the medium. The aggregated complexes did not require proteasomal degradation for clearance. Aggregate formation was enhanced by ATP depletion, and upon restoration of metabolic energy, these aggregates were dissociated and secreted. With the coexpression of von Willebrand factor (vWF), a small portion of vWF coaggregated with factor VIII. However, vWF dissociated from the aggregates more rapidly than factor VIII, supporting that these aggregates are dynamic. An increase in the factor VIII expression level elicited a corresponding increase in the fraction of factor VIII that was aggregated. In addition, a 110 amino acid sequence containing a hydrophobic beta-sheet within factor VIII was identified that may predispose factor VIII to aggregation. These data show that formation and ATP dependent dissolution of nondisulfide-linked factor VIII aggregates is a dynamic, rate-limiting step during the folding process in the early secretory pathway. In summary, we have identified an unprecedented requirement for protein transport out of the ER that involves an ATP-dependent dissociation of non-disulfide-linked aggregates within the ER. PMID- 10684648 TI - Competition of annexin V and anticardiolipin antibodies for binding to phosphatidylserine containing membranes. AB - Annexin V, an intracellular protein with a calcium-dependent high affinity for anionic phospholipid membranes, acts as an inhibitor of lipid-dependent reactions of the blood coagulation. Antiphospholipid antibodies found in the plasma of patients with antiphospholipid syndrome generally do not interact with phospholipid membranes directly, but recognize (plasma) proteins associated with lipid membranes, mostly prothrombin or beta(2)-glycoprotein I (beta(2)GPI). Previously, it has been proposed that antiphospholipid antibodies may cause thrombosis by displacing annexin V from procoagulant cell surfaces. We used ellipsometry to study the binding of annexin V and of complexes of beta(2)GPI with patient-derived IgG antibodies to beta(2)GPI, commonly referred to as anticardiolipin antibodies (ACA), to phospholipid bilayers composed of phosphatidylcholine (PC) and 20% phosphatidylserine (PS). More specifically, we investigated the competition of these proteins for the binding sites at these bilayers. We show that ACA-beta(2)GPI complexes, adsorbed to PSPC bilayers, are displaced for more than 70% by annexin V and that annexin V binding is unaffected by the presence of ACA-beta(2)GPI complexes. Conversely, annexin V preadsorbed to these bilayers completely prevents adsorption of ACA-beta(2)GPI complexes, and none of the preadsorbed annexin V is displaced by ACA-beta(2)GPI complexes. Using ellipsometry, we also studied the effect of ACA-beta(2)GPI complexes on the interaction of annexin V with the membranes of ionophore-activated blood platelets as a more physiological relevant model of cell membranes. The experiments with blood platelets confirm the high-affinity binding of annexin V to these membranes and unequivocally show that annexin V binding is unaffected by the presence of ACA-beta(2)GPI. In conclusion, our data unambiguously show that ACA-beta(2)GPI complexes are unable to displace annexin V from procoagulant membranes to any significant extent, whereas annexin V does displace the majority of preadsorbed ACA-beta(2)GPI complexes from these membranes. PMID- 10684649 TI - C-terminal domains of Na(+)/H(+) exchanger isoform 3 are involved in the basal and serum-stimulated membrane trafficking of the exchanger. AB - When expressed either in polarized epithelial cells or in fibroblasts, two Na(+)/H(+) exchanger isoforms, NHE1 and NHE3, have different subcellular distributions. Using a quantitative cell surface biotinylation technique, we found PS120 cells target approximately 90% of mature NHE1 but only 14% of NHE3 to the cell surface, and this pattern occurs irrespective of NHE protein expression levels. In this study, we examined surface fractions of NHE3 C-terminal truncation mutants to identify domains involved in the targeting of NHE3. Removing the C-terminal 76 amino acids doubled surface fractions to 30% of total and doubled the V(max) from 1300 to 2432 microM H(+)/s. Removal of another 66 amino acids increased surface levels to 55% of total with an increase in the V(max) to 5794 microM H(+)/s. Surface fractions did not change with a further 105 amino acid truncation. We postulated that inhibition of the basal recycling of NHE3 could result in the surface accumulation of the NHE3 truncations. Accordingly, we found that, unlike wild-type NHE3, the truncations were shown to internalize poorly and were not affected by PI3 kinase inhibition. However, while the truncations demonstrated reduced basal recycling, they retained the same serum response as full-length NHE3, with a mobilization of approximately 10% of total NHE to the surface. We conclude that basal recycling of NHE3 is controlled by endocytic determinants contained within its C-terminal 142 amino acids and that serum-mediated exocytosis is independently regulated through a different part of the protein. PMID- 10684650 TI - Structure-activity relationships for the interaction of bovine pancreatic trypsin inhibitor with an intracellular site on a large conductance Ca(2+)-activated K(+) channel. AB - Large conductance Ca(2+)-activated K(+) channels (BK(Ca)) contain an intracellular binding site for bovine pancreatic trypsin inhibitor (BPTI), a well known inhibitor of various serine proteinase (SerP) enzymes. To investigate the structural basis of this interaction, we examined the activity of 11 BPTI mutants using single BK(Ca) channels from rat skeletal muscle incorporated into planar lipid bilayers. All of the mutants induced discrete substate events at the single channel level. The dwell time of the substate, which is inversely related to the dissociation rate constant of BPTI, exhibited relatively small changes (<9-fold) for the various mutants. However, the apparent association rate constant varied up to 190-fold and exhibited a positive correlation with the net charge of the molecule, suggesting the presence of a negative electrostatic surface potential in the vicinity of the binding site. The substate current level was unaffected by most of the mutations except for substitutions of Lys15. Different residues at this position were found to modulate the apparent conductance of the BPTI-induced substate to 0% (K15G), 10% (K15F), 30% (K15 wild-type), and 55% (K15V) of the open state at +20 mV. Lys15 is located on a loop of BPTI that forms the primary contact region for binding to many SerPs such as trypsin, chymotrypsin, and elastase. The finding that Lys15 is a determinant of the conductance behavior of the BK(Ca) channel when BPTI is bound implies that the same inhibitory loop that contacts SerP's is located close to the protein interface in the BK(Ca) channel complex. This supports the hypothesis that the C-terminal region of the BK(Ca) channel protein contains a domain homologous to SerP's. We propose a domain interaction model for the mechanism of substate production by Kunitz inhibitors based on current ideas for allosteric activation of BK(Ca) channels by voltage and Ca(2+). PMID- 10684651 TI - The Leishmania GDP-mannose transporter is an autonomous, multi-specific, hexameric complex of LPG2 subunits. AB - LPG2 (a gene involved in lipophosphoglycan assembly) encodes the Golgi GDP-Man transporter of the protozoan parasite Leishmania and is a defining member of a new family of eukaryotic nucleotide-sugar transporters (NSTs). Although NST activities are widespread, mammalian cells lack a GDP-Man NST, thereby providing an ideal heterologous system for probing the LPG2 structure and activity. LPG2 expression constructs introduced into either mammalian cells or a Leishmania lpg2(-) mutant conferred GDP-Man, GDP-Ara, and GDP-Fuc (in Leishmania only) uptake in isolated microsomes. LPG2 is the first NST to be associated with multiple substrate specificities. Uptake activity showed latency, exhibited an antiport mechanism of transport with GMP, and was susceptible to the anion transport inhibitor DIDS. The apparent K(m) for GDP-Man uptake was similar in transfected mammalian cells (12.2 microM) or Leishmania (6.9 microM). Given the evolutionary distance between protozoans and vertebrates, these data suggest that LPG2 functions autonomously to provide transporter activity. Using epitope-tagged LPG2 proteins, we showed the existence of hexameric LPG2 complexes by immunoprecipitation experiments, glycerol gradient centrifugation, pore-limited native gel electrophoresis, and cross-linking experiments. This provides strong biochemical evidence for a multimeric complex of NSTs, a finding with important implications to the structure and specificity of NSTs in both Leishmania and other organisms. Inhibition of essential GDP-Man uptake in fungal and protozoan systems offers an attractive target for potential chemotherapy. PMID- 10684652 TI - Cellular phosphorylation of an acidic proline-rich protein, PRP1, a secreted salivary phosphoprotein. AB - Phosphorylation of many secreted salivary proteins is necessary for their biological functions. Identification of the kinase, which is responsible for in vivo phosphorylation, is complicated, because several of the protein phosphorylation sites conform both to the recognition sequence of casein kinase 2 (CK2) and Golgi kinase (G-CK), which both are found in the secretory pathway. This study was undertaken to determine the kinase recognition sequence in a secreted proline-rich salivary protein, PRP1, and thereby identify the responsible kinase. This was done by transfecting a human submandibular cell line, HSG, and a kidney cell line, HEK293, with expression vectors encoding wild type or mutated PRP1. It was shown that phosphorylation occurred only at the same sites, Ser8 and 22, as in PRP1 purified from saliva. Phosphorylation at either site did not depend on the other site being phosphorylated. The sequence surrounding Ser8 has characteristics of both CK2 and G-CK recognition sequences, but destruction of the CK2 recognition site had no effect on phosphorylation, whereas no phosphorylation occurred if the G-CK recognition sequence was altered. The sequence surrounding Ser22 did not conform to any known kinase recognition sites. If Ser22 was mutated to Thr, no phosphorylation was seen, and a cluster of negatively charged residues at positions 27-29 was identified as part of the enzyme recognition site. Ser22 may be phosphorylated by a G-CK that recognizes an atypical substrate sequence or by a novel kinase. No difference in phosphorylation was seen between undifferentiated and differentiated HSG cells. PMID- 10684653 TI - Recovery of photosystem II activity in photoinhibited synechocystis cells: light dependent translation activity is required besides light-independent synthesis of the D1 protein. AB - Irreversible photoinactivation of photosystem II (PSII) results in the degradation of the reaction center II D1 protein. In Synechocystis PCC 6714 cells, recovery of PSII activity requires illumination. The rates of photoinactivation and recovery of PSII activity in the light are similar in cells grown in minimal (MM) or glucose-containing medium (GM). Reassembly of PSII with newly synthesized proteins requires degradation of the D1 protein of the photoinactivated PSII. This process may occur in darkness in both types of cells. The degraded D1 protein is, however, only partially replaced by newly synthesized protein in MM cells in darkness while a high level of D1 protein synthesis occurs in darkness in the GM cells. The newly synthesized D1 protein in darkness appears to be assembled with other PSII proteins. However, PSII activity is not recovered in such cells. Illumination of the cells in absence but not in the presence of protein synthesis inhibitors allows recovery of PSII activity. PMID- 10684654 TI - Rebinding of IgE Fabs at haptenated planar membranes: measurement by total internal reflection with fluorescence photobleaching recovery. AB - In previous work, a general analytical theory for ligand rebinding at cell surfaces was developed for a reversible bimolecular reaction between ligands in solution and receptors on a membrane surface [Lagerholm, B. C., and Thompson, N. L. (1998) Biophys. J. 74, 1215-1228]. This theory can be used to predict theoretical forms for data obtained by using total internal reflection with fluorescence photobleaching recovery (TIR-FPR) [Thompson, N. L., Burghardt, T. P., and Axelrod, D. (1981) Biophys. J. 33, 435-454]. Thus, one method by which the rebinding theory can be tested is to use TIR-FPR. In the work described herein, the reversible kinetics of mouse monoclonal anti-dinitrophenyl (DNP) IgE Fabs at substrate-supported planar membranes composed of 25 mol % DNP-conjugated phosphatidylethanolamine and 75 mol % dipalmitoylphosphatidylcholine have been examined by using TIR-FPR. Data were obtained as a function of the Fab solution concentration. Higher Fab concentrations reduce rebinding (and increase the fluorescence recovery rate) because different Fab molecules compete for the same surface-binding sites. Data were also obtained for solutions containing different volume fractions of glycerol. In these measurements, higher glycerol concentrations increase rebinding (and decrease the fluorescence recovery rate) because the solution viscosity is increased and the Fab diffusion coefficient in solution is decreased. The TIR-FPR data were quantitatively compared with theoretical predictions which follow from the general theory for rebinding at the membrane surface. The data were consistent with the theoretical predictions and, therefore, provide experimental verification of the previously developed theory. PMID- 10684655 TI - From redox flow to gene regulation: role of the PrrC protein of Rhodobacter sphaeroides 2.4.1. AB - Activation of photosynthesis (PS) gene expression by the PrrBA two-component activation system in Rhodobacter sphaeroides 2.4.1 results from the interruption of an inhibitory signal originating from the cbb(3) cytochrome c oxidase via its interaction with oxygen, in conjunction with the Rdx redox proteins. The CcoQ protein, encoded by the ccoNOQP operon, which encodes the cbb(3) cytochrome c oxidase, was shown to act as a "transponder" that conveys the signal derived from reductant flow through cbb(3) to oxygen, to the Prr system. To further define the elements comprising this signal transduction pathway we considered the prrC gene product, which to date possessed no definable role in this signal transduction pathway despite its being part of the prrBCA gene cluster. Similar to mutations in cbb(3) and rdx, suitably constructed prrC deletion mutations lead to PS gene expression in the presence of high oxygen. Unlike mutations that remove cbb(3) terminal oxidase activity or Rdx function, the PrrC deletion mutant shows no effect upon cbb(3) activity, nor does it affect the ratio of the carotenoid (Crt) spheroidene (SE) to spheroidenone (SO). Thus, the PrrC deletion mutant behaves identically to the CcoQ deletion mutant. Taking these and previous results together, we suggest that PrrC is located upstream of the two-component PrrBA activation system in the signal transduction pathway but downstream of the cbb(3) cytochrome c oxidase and its "transponder" CcoQ. The PrrC deletion mutant was also shown to lead to an increase in the DorA protein under aerobic conditions as was shown earlier for the cbb(3) mutant. Finally, PrrC is a member of a highly conserved family of proteins found in both prokaryotes and eukaryotes, and this appears to be the first instance in which a direct regulatory role has been ascribed to a member of this protein family. PMID- 10684656 TI - A new photolabile precursor of glycine with improved properties: A tool for chemical kinetic investigations of the glycine receptor. AB - The synthesis and characterization of a new photolabile precursor of glycine (caged glycine) is described. The alpha-carboxyl group of glycine is covalently coupled to the alpha-carboxy-2-nitrobenzyl (alphaCNB) protecting group. Photolysis of the caged glycine with UV light produces free glycine. At 308 nm, the compound photolyzes with a quantum yield of 0.38. The absorption spectrum and the pH dependence of a transient absorption produced after laser-flash illumination are typical for aci-nitro intermediates of alphaCNB-protected compounds. The time constant for the major component of the aci-nitro intermediate decay ( approximately 84% of the total aci-nitro absorbance) was determined to be 7 micros at physiological pH. A minor component ( approximately 16%) decays with a rate constant of 170 micros. The compound does not activate or inhibit the alpha(1)-homomeric glycine receptor transiently expressed in HEK293 cells. After photolysis with a 10 ns pulse of 325 nm laser light, the glycine released from the caged compound activates glycine-mediated whole-cell currents in the same cells. The rise of these currents can be measured in a time-resolved fashion and occurs on a millisecond to sub-millisecond time scale. It can be described with a single-exponential function over >85% of the total current. The rate constant of the current rise is about 2 orders of magnitude slower than the rate constant of caged glycine photolysis. Thermal hydrolysis of the alphaCNB caged glycine takes place with a half-life of 15.6 h at physiological pH. The new caged glycine is the first in a series of photoprotected glycine derivatives that has the required properties for use with chemical kinetic methods for investigation of glycine-activated cell surface receptors. Photolysis is rapid and efficient with respect to the receptor reactions to be studied; hydrolysis in aqueous solution is sufficiently slow, and the compound is biologically inert. It will, therefore, be a useful tool for investigation of the processes leading to channel opening of glycine receptor channels and the effects of mutations of the glycine receptor and of inhibitors on these processes. PMID- 10684657 TI - Effects of aromatic residues at the ends of transmembrane alpha-helices on helix interactions with lipid bilayers. AB - We have studied the effects of aromatic residues at the ends of peptides of the type Ac-KKGL(n)()WL(m)()KKA-amide on their interactions with lipid bilayers as a function of lipid fatty acyl chain length, physical phase, and charge. Peptide Ac KKGFL(6)WL(8)FKKA-amide (F(2)L(14)) incorporated into bilayers of phosphatidylcholines containing monounsaturated fatty acyl chains of lengths C14 C24 at a peptide:lipid molar ratio of 1:100 in contrast to Ac-KKGL(7)WL(9)KKA amide (L(16)) which did not incorporate at all into dierucoylphosphatidylcholine [di(C24:1)PC]; Ac-KKGYL(6)WL(8)YKKA-amide (Y(2)L(14)) incorporated partly into di(C24:1)PC. Lipid-binding constants relative to that for dioleoylphosphatidylcholine (C18:1)PC were obtained using a fluorescence quenching method. For Y(2)L(14) and F(2)L(14), relative lipid-binding constants increased with increasing fatty acyl chain length from C14 to C24; strongest binding did not occur at the point where the hydrophobic length of the peptide equalled the hydrophobic thickness of the bilayer. For Ac-KKGYL(9)WL(11)YKKA amide (Y(2)L(20)), increasing chain length from C18 to C24 had little effect on relative binding constants. Anionic phospholipids bound more strongly than zwitterionic phospholipids to Y(2)L(14) and Y(2)L(20) but effects of charge were relatively small. In two phase (gel and liquid crystalline) mixtures, all the peptides partitioned more strongly into liquid crystalline than gel phase; effects were independent of the structure of the peptide or of the lipid (dipalmitoylphosphatidylcholine or bovine brain sphingomyelin). Addition of cholesterol had little effect on incorporation of the peptides into lipid bilayers. It is concluded that the presence of aromatic residues at the ends of transmembrane alpha-helices effectively buffers them against changes in bilayer thickness caused either by an increase in the chain length of the phospholipid or by the presence of cholesterol. PMID- 10684658 TI - Kinetic mechanism of the p38-alpha MAP kinase: phosphoryl transfer to synthetic peptides. AB - p38 is a member of the mitogen-activated protein (MAP) kinase family. Activation (phosphorylation) of p38 acts as a switch for the transcriptional and translational regulation of a number of proteins, including the proinflammatory cytokines. Investigation of a set of small peptides revealed that, as with protein substrates, p38-alpha behaves as a proline-directed Ser/Thr MAP kinase for a peptide substrate, peptide 4 (IPTSPITTTYFFFKKK). We investigated the steady state kinetic mechanism of the p38-alpha-catalyzed kinase reaction with EGF receptor peptide, peptide 1, as a substrate. Lineweaver-Burk analysis of the substrate kinetics yielded a family of lines intersecting to the left of the ordinate, with either ATP or peptide 1 as the varied substrate. Kinetic analysis in the presence of ADP yielded a competitive inhibition pattern when ATP was the varied substrate and a noncompetitive pattern if peptide 1 was the varied substrate. At saturating peptide substrate concentrations, inhibition by phosphopeptide product yielded an uncompetitive pattern when ATP was the varied substrate. These data are consistent with ordered binding with ATP as the initial substrate. We provide further evidence of the existence of a productive p38.ATP binary complex in that (a) activated p38-alpha has intrinsic ATPase activity, (b) ATPase and kinase activities are coupled, and (c) inhibitors of ATPase activity also inhibit the kinase activity with a similar inhibition constant. The k(cat) for the kinase reaction was lowered by 1.8-fold when ATP-gamma-S was used. Microviscosity linearly affected the k(cat) values of both the ATP and ATP-gamma S reactions with a slope of about 0.8. These observations were interpreted to mean that the phosphoryl transfer step is not rate-limiting and that the release of product and/or enzyme isomerization is a possible rate-limiting step(s). PMID- 10684659 TI - Mechanistic analysis of a type II polyketide synthase. Role of conserved residues in the beta-ketoacyl synthase-chain length factor heterodimer. AB - Type II polyketide synthases (PKSs) are a family of multienzyme systems that catalyze the biosynthesis of polyfunctional aromatic natural products such as actinorhodin, frenolicin, tetracenomycin, and doxorubicin. A central component in each of these systems is the beta-ketoacyl synthase-chain length factor (KS-CLF) heterodimer. In the presence of an acyl carrier protein (ACP) and a malonyl CoA:ACP malonyl transferase (MAT), this enzyme synthesizes a polyketide chain of defined length from malonyl-CoA. We have investigated the role of the actinorhodin KS-CLF in priming, elongation, and termination of its octaketide product by subjecting the wild-type enzyme and selected mutants to assays that probe key steps in the overall catalytic cycle. Under conditions reflecting steady-state turnover of the PKS, a unique acyl-ACP intermediate is detected that carries a long, possibly full-length, acyl chain. This species cannot be synthesized by the C169S, H309A, K341A, and H346A mutants of the KS, all of which are blocked in early steps in the PKS catalytic cycle. These four residues are universally conserved in all known KSs. Malonyl-ACP alone is sufficient for kinetically and stoichiometrically efficient synthesis of polyketides by the wild type KS-CLF, but not by heterodimers that carry the mutations listed above. Among these mutants, C169S is an efficient decarboxylase of malonyl-ACP, but the H309A, K341A, and H346A mutants are unable to catalyze decarboxylation. Transfer of label from [(14)C]malonyl-ACP to the nucleophile at position 169 in the KS can be detected for the wild-type enzyme and for the C169S and K341A mutants, but not for the H309A mutant and only very weakly for the H346A mutant. A model is proposed for decarboxylative priming and extension of a polyketide chain by the KS, where C169 and H346 form a catalytic dyad for acyl chain attachment, H309 positions the malonyl-ACP in the active site and supports carbanion formation by interacting with the thioester carbonyl, and K341 enhances the rate of malonyl ACP decarboxylation via electrostatic interaction. Our data also suggest that the ACP and the KS dissociate after each C-C bond forming event, and that the newly extended acyl chain is transferred back from the ACP pantetheine to the KS cysteine before dissociation can occur. Chain termination is most likely the rate limiting step in polyketide biosynthesis. Within the act CLF, neither the universally conserved S145 residue nor Q171, which aligns with the active site cysteine of the ketosynthase, is essential for PKS activity. The results described here provide a basis for a better understanding of the catalytic cycle of type II PKSs and fatty acid synthases. PMID- 10684660 TI - tRNA 3' processing in plants: nuclear and mitochondrial activities differ. AB - The nuclear tRNA 3' processing activity from wheat has been characterized and partially purified. Several characteristics of the wheat nuclear 3' processing enzyme now allow this activity to be distinguished from its mitochondrial counterpart. The nuclear enzyme is an endonuclease, which we termed nuclear RNase Z. The enzyme cleaves at the discriminator base and seems to consist only of protein subunits, since essential RNA subunits could not be detected. RNase Z leaves 5' terminal phosphoryl and 3' terminal hydroxyl groups at the processing products. It is a stable enzyme being active over broad temperature and pH ranges, with the highest activity at 35 degrees C and pH 8.4. The apparent molecular mass according to gel filtration chromatography is 122 kDa. The nuclear RNase Z does process 5' extended pretRNAs but with a much lower efficiency than 5' matured pretRNAs. Nuclear intron-containing precursor tRNAs as well as mitochondrial precursor tRNAs are efficiently cleaved by the nuclear RNase Z. Mitochondrial pretRNA(His) is processed by the nuclear RNase Z, generating a mature tRNA(His) containing an 8 base pair acceptor stem. The edited mitochondrial pretRNA(Phe) is cleaved easily, while the unedited version having a mismatch in the acceptor stem is not cleaved. Thus, an intact acceptor stem seems to be required for processing. Experiments with precursors containing mutated tRNAs showed that a completely intact anticodon arm is not necessary for processing by RNase Z. Comparison of the plant nuclear tRNA 3' processing enzyme with the plant mitochondrial one suggests that both activities are different enzymes. PMID- 10684661 TI - Kinetic mechanism of nucleotide cofactor binding to Escherichia coli replicative helicase DnaB protein. stopped-flow kinetic studies using fluorescent, ribose-, and base-modified nucleotide analogues. AB - The kinetic mechanism of binding nucleotide cofactors to the Escherichia coli primary replicative helicase DnaB protein has been studied, using the fluorescence stopped-flow technique. The experiments have been performed with fluorescent ATP and ADP analogues bearing the modification on the ribose, MANT AMP-PNP and MANT-ADP, and on the base, epsilonAMP-PNP and epsilonADP. Association of the DnaB helicase with nucleotide cofactors is characterized by four relaxation times that indicate that the binding occurs by a minimum of four steps. The simplest mechanism which can describe the data is a four-step sequential process where the bimolecular binding step is followed by three isomerization steps. This mechanism is described by the following equation: [equation in text]. The binding mechanism is independent of the location of the nucleotide cofactor modification and is an intrinsic property of the DnaB helicase-nucleotide system. Quantitative amplitude analyses, using the matrix projection operator technique, allowed us to determine specific fluorescence changes accompanying the formation of all intermediates relative to the fluorescence of the free nucleotide. It shows that the major conformational change of the DnaB helicase-nucleotide complex occurs in the formation of the (H N)(1). Moreover, the value of the bimolecular rate constant, k(1), is 3-4 orders of magnitude lower than the value expected for the diffusion-controlled reaction. These results indicate that the determined first step includes formation of the collision and an additional transition of the enzyme-nucleotide complex. The obtained results provide evidence of profoundly different conformational states of the ribose and base regions of the nucleotide-binding site in different intermediates. The sequential nature of the mechanism of the nucleotide binding to the DnaB helicase indicates the lack of the existence of a kinetically significant conformational equilibrium of the helicase protomer and the DnaB hexamer prior to the binding. The significance of these results for the functioning of the DnaB helicase is discussed. PMID- 10684663 TI - A cyclophilin-regulated PP2A-like protein phosphatase in thylakoid membranes of plant chloroplasts PMID- 10684664 TI - Electroviscous Forces on a Charged Cylinder Moving Near a Charged Wall. AB - The refined theory of the electroviscous lift forces is presented for the case when the separation distance between the particle and the wall is larger than the double-layer thickness. The theory is based on the lubrication approximation for motion of a long cylinder near a solid wall in creeping flow. The approximate analytical formula for the lift force valid for Pe50%, palpable positive pelvic nodes, or positive adnexae), excluding stage IV disease, and (2) 51 patients with positive nodes (pelvic or para aortic), excluding stage IV disease. By our definition, PAL required removal of five or more para-aortic nodes. RESULTS: In both groups, no significant difference existed between patients who had PAL (PAL+) and those who did not (PAL ) in regard to clinical or pathologic variables, percentage irradiated, or surgical or radiation complications. Among the 137 high-risk patients, the 5-year progression-free survival was 62% and the 5-year overall survival was 71% for the PAL- group compared with 77 and 85%, respectively, for the PAL+ group (P = 0.12 and 0.06, respectively). For the 51 patients with positive nodes, the 5-year progression-free survival and 5-year overall survival for the PAL- group were 36 and 42% compared with 76 and 77% for the PAL+ group (P = 0.02 and 0.05, respectively). Lymph node recurrences were detected in 37% of the PAL- patients but in none of the PAL+ patients (P = 0.01). Multivariate analysis suggested that submission to PAL was a cogent predictor of progression-free survival (odds ratio = 0.25; P = 0.01) and overall survival (odds ratio = 0.23; P = 0.006). CONCLUSIONS: These results suggest a potential therapeutic role for formal PAL in endometrial cancer. PMID- 10684710 TI - Clinical value of intraoperative gross examination in endometrial cancer. AB - We present the largest multicenter study evaluating whether intraoperative visual estimation can accurately assess the depth of myometrial invasion in patients with endometrial cancer. The study population consisted of 403 consecutive women who underwent total hysterectomy for endometrial cancer. After the uterus was removed, a visual estimate of depth of gross myometrial invasion was recorded. The uterus was opened, the endometrial cavity was inspected, and one or more full thickness incisions were made through the tumor, myometrium, and serosa. An intraoperative estimation of gross myometrial invasion was made and classified as more or less than 50% of the uterine wall. Gross visual estimation accurately identified the microscopic myometrial invasion in 85.3% (344/403) of cases. Sensitivity, specificity, and positive and negative predictive values of gross estimation in determining a microscopic myometrial invasion greater than 50% were 73.0, 92.5, 85.0, and 85.5%, respectively. Among patients in whom the myometrial invasion was underestimated at gross examination the tumoral invasion was limited to the inner two thirds of the myometrium in 45% (18/40) of cases and the distance from the tumor-myometrial junction to the uterine serosa was greater than 3 mm in 65% (26/40) of cases. We conclude that gross estimation of myometrial invasion is a reliable and inexpensive method for evaluating the invasiveness of uterine carcinomas and that deciding to perform an extensive surgical staging upon gross estimation will be in accordance with the final histopathologic report in about 9 of 10 cases. PMID- 10684711 TI - Development of a panel of 15 human ovarian cancer xenografts for drug screening and determination of the role of the glutathione detoxification system. AB - OBJECTIVES: We have established a panel of 15 human ovarian cancer xenografts grown subcutaneously in the flank of the nude mouse. Similar to the clinic, the xenografts show differences in histological subtype and volume doubling time. We determined whether the panel is useful for drug screening by testing the sensitivity to six conventional anticancer agents. In addition, we investigated whether the glutathione detoxification system affects sensitivity to cisplatin and cyclophosphamide, major drugs in the treatment of ovarian cancer. METHODS: Mice bearing well-established tumors were treated at maximum tolerated doses as defined by a reversible weight loss up to 15% of their initial weight: cisplatin 5 mg/kg iv weekly x2, cyclophosphamide 150 mg/kg ip 2-weekly x2, doxorubicin 8 mg/kg iv weekly x2, hexamethylmelamine ip 150 mg/kg every other day x4, methotrexate ip 150 mg/kg weekly x2, and 5-fluorouracil 60 mg/kg ip weekly x4. Glutathione levels and the activities of three different glutathione-dependent enzymes were measured in untreated xenograft tissues. RESULTS: Growth inhibition >75% was reached for cisplatin in 40%, for cyclophosphamide in 27%, and for doxorubicin in 20% of the xenografts. Methotrexate and 5-fluorouracil did not induce growth inhibition of importance. Hexamethylmelamine showed >75% growth inhibition in 53% of the xenografts, which may have been caused by the favorable metabolism of the drug in mice when compared with that in patients. Glutathione levels varied 3.6-fold in the xenografts and did not show a relation with sensitivity to cisplatin, cyclophosphamide, or doxorubicin. No relation was found between the activities of glutathione S-transferase and glutathione peroxidase and the sensitivities to the three anticancer agents. Glutathione reductase activity, however, showed a weak, inverse relation with the efficacy of cisplatin and cyclophosphamide (r values of -0.55 and -0.58, respectively). CONCLUSIONS: The sensitivity to the six anticancer agents of our panel of 15 human ovarian cancer xenografts reflects the response rates known for similar drugs in ovarian cancer patients. In that respect, the panel may be useful for drug screening as well as studies on the relevance of drug resistance features in vivo. The various components of the glutathione detoxification system did not predict for primary drug resistance which confirms clinical data in ovarian cancer. PMID- 10684712 TI - Genetic alterations of the WT1 gene in papillary serous carcinoma of the peritoneum. AB - OBJECTIVE: The Wilms' tumor (WT1) gene product is consistently detectable in both normal ovarian germinal epithelium and human mesothelium. Ovarian carcinomas frequently exhibit alterations in WT1 function. Papillary serous carcinoma of the peritoneum (PSCP) is believed to develop de novo from the peritoneal lining (mesothelium) of the pelvis and abdomen. The purpose of this study was to determine if genetic alterations of the WT1 gene are associated with the development of PSCP. METHODS: Normal and tumor tissue specimens were retrieved from patients with stage III and IV PSCP (n = 38) and serous epithelial ovarian carcinoma (n = 38). Immunohistochemistry was performed using the anti-WT1 (C-19) antibody. Loss of heterozygosity (LOH) was performed at the WT1 locus. Clinical data were obtained and correlated with molecular findings. RESULTS: Loss of normal WT1 expression was detected in 18 (51%) of 35 PSCP specimens and 18 (53%) of 34 ovarian carcinoma specimens. Six (27%) of 22 PSCP specimens and 3 (13%) of 24 ovarian carcinoma specimens had LOH at the WT1 locus (P = 0.27). Normal WT1 gene expression was maintained in 86% of tumors exhibiting LOH. Genetic alterations of the WT1 gene were not predictive of survival, nor were they associated with other clinical or molecular factors. CONCLUSIONS: Genetic alterations of the WT1 gene are associated with the development of PSCP. The loss of normal WT1 gene expression is a common event in both PSCP and advanced ovarian carcinoma, likely resulting from down-regulation by other regulatory factors-not from inactivating gene mutation and subsequent allelic loss. PMID- 10684713 TI - The prognostic significance of radiation dose and residual tumor in the treatment of barrel-shaped endophytic cervical carcinoma. AB - OBJECTIVE: The aim of this study was to evaluate the impact of total radiation dose on residual tumor and the prognostic significance of persistent disease in women with bulky, barrel-shaped cervical carcinoma who received definitive radiation followed by adjuvant hysterectomy. METHODS: The medical records of 57 patients with bulky endophytic cervical carcinoma treated at the University of Washington between 1976 and 1997 were reviewed. All patients received external beam pelvic radiotherapy supplemented by intracavitary brachytherapy, followed by extrafascial hysterectomy 6 to 8 weeks later. RESULTS: The mean pretreatment tumor diameter was 5.9 cm, with a range of 4-9 cm. Total radiation dose to point A ranged from 5040 to 9700 cGy, and the mean for the group was 7966 cGy. Residual disease was present in 35 (61%) of the hysterectomy specimens. The frequency of cervical tumor sterilization correlated significantly with the mean radiation dose to point A (P = 0.016). Patients without histologic residual disease had a significantly improved outcome, with 95% of patients remaining clinically free of disease at last follow-up, versus 31% of those with residual disease (P < 0.001). As expected, the pelvic control rate was excellent (100%) in patients with complete tumor eradication compared to the group with residual tumor (44%). Those with no residual disease enjoyed a significantly improved survival compared to those with residual tumor (P < 0.001). Furthermore, a statistically significant higher survival was realized in patients harboring only microscopic residual compared to those with either macroscopically evident tumor residuum and/or positive surgical margins (P = 0.036). CONCLUSIONS: Higher radiation doses are associated with an improved likelihood of tumor eradication in the treatment of bulky, endophytic cervical cancer and complete tumor sterilization at adjuvant hysterectomy is predictive of significantly enhanced survival and pelvic control. The high rate of histologic tumor persistence in our series emphasizes the need for more efficacious therapies in patients with bulky endophytic cervical cancer and argues for escalation of radiation dose even when adjuvant hysterectomy is planned. PMID- 10684714 TI - Laparoscopic loop colostomy for advanced ovarian cancer, rectal cancer, and rectovaginal fistulas. AB - OBJECTIVES: The objectives of this study were to present a minimal invasive technique of intestinal diversion for selected cases of advanced inoperable ovarian cancer, rectal cancer, and rectovaginal fistulas, and to discuss indications, pitfalls, and potential complications. METHODS: The technical features of laparoscopic colostomy are described. RESULTS: Between August 1995 and July 1997 laparoscopic colostomy was successfully carried out in 12 patients with advanced ovarian cancer, inoperable carcinoma of the rectum, or rectovaginal fistulas. There were no intraoperative or postoperative complications and postoperative recovery was rapid, with all patients having function of the colostomy within 24 h and regaining their preoperative state of mobility on the second postoperative day. CONCLUSION: The laparoscopic approach allows careful selection of the colostomy site and easy mobilization of the colon, causing only little disruption to intestinal function and, hence, improving postoperative recovery. From our experience, laparoscopic colostomy is in most cases a simple and safe operation and can be used as the preferred technique of intestinal diversion. PMID- 10684715 TI - Assessing disease extent in women with bulky or clinically evident metastatic cervical cancer: yield of pretreatment studies. AB - OBJECTIVE: The objective of this study was to determine the impact of pretreatment tests on staging and management for women with bulky or clinically evident metastatic cervical cancer. METHODS: Demographics and findings of clinic pelvic examination (PE), examination under anesthesia (EUA), chest x ray (CXR), abdominopelvic computed tomography (CT), and intravenous urography (IVU) were reviewed for women with primary, untreated cervical cancers either > or =4 cm or with extracervical disease apparent on PE or CXR evaluated between July 1, 1994, and March 31, 1999. Stage was assigned according to standards of the International Federation of Gynecology and Obstetrics. RESULTS: In 133 women studied, tumor diameter averaged 4. 9 cm on PE and 5.7 cm at EUA (P = 0.0005). Of 92 women undergoing both PE and EUA, 30 (33%) had size discrepancies of more than 1 cm. Compared with EUA, PE had sensitivity, specificity, and positive and negative predictive values, respectively, of 65, 89, 79, and 81% for vaginal disease, 74, 91, 95, and 63% for parametrial disease, and 57, 90, 60, and 89% for sidewall fixation. CXR was abnormal in 5 (4%). IVU showed ureteral dilation in 20 (22%) of 90 women, while CT showed dilation in 34 (28%) of 123 women. CT also showed pelvic lymphadenopathy in 22 (18%) women and paraaortic lymphadenopathy in 12 (10%). Bladder biopsies showed malignancy in 8 (8%), including one transitional cell carcinoma of the bladder. Proctoscopy failed to reveal metastatic cervical cancer. CONCLUSIONS: CXR and EUA with cystoscopy are important to the accurate staging and treatment of bulky or clinically metastatic cervical cancers, while proctoscopy is rarely useful. CT predicts ureteral obstruction well, and its greater information yield may justify its higher cost compared with IVU. PMID- 10684716 TI - Expression of fos and jun proto-oncogenes in benign versus malignant human uterine tissue. AB - OBJECTIVE: The objective of this study was to evaluate expression of fos and jun proto-oncogenes in benign human uterine tissue compared with malignant uterine tissue. METHODS: Forty-two endometrial tissue specimens were obtained at the time of hysterectomy. Tissue samples from different phases of the menstrual cycle and from postmenopausal patients were stained using immunohistochemical methods to detect Fos and Jun proteins, estrogen and progesterone receptor status, and Ki67 (detects a nuclear antigen associated with proliferating cells). Tissue was examined microscopically for nuclear staining in endometrial epithelium and stroma. The endometrium was based on the patient's last menstrual period, pathologic dating, and proliferative versus nonproliferative status as determined by Ki67. Benign and malignant specimens were subjected to Northern blot analysis to evaluate levels of expression of c-fos, c-jun, and jun-B mRNA. The pattern of c-fos mRNA expression in malignant samples was further evaluated using in situ hybridization. RESULTS: In proliferative, secretory, postmenopausal, and progesterone-influenced, uterine specimens immunohistochemically stained and examined, the endometrial and stromal nuclei stained for both Fos and Jun in varying intensities. However, no pattern was found in the variation of intensity according to the phase of the endometrium. Similarly, in malignant and benign endometrial tissue examined by Northern blot and in situ hybridization analyses, expression of proto-oncogene mRNAs was readily detectable, but no statistical correlation between type of tissue examined, grade of adenocarcinoma, and stage of endometrial cancer was found in this study. CONCLUSIONS: In rodent models, control of uterine cell proliferation is related to change in expression of fos and jun proto-oncogenes. Our results indicate that hormonal control is likely to be different in human endometrium and probably involves genes other than the proto-oncogenes under study. Expression of Fos and Jun do not correlate with endometrial cancer stage and grade. PMID- 10684717 TI - Intraperitoneal photoimmunotherapy of ovarian carcinoma xenografts in nude mice using charged photoimmunoconjugates. AB - OBJECTIVE: The objective of this study was to compare the efficacy of photoimmunoconjugates with cationic and anionic molecular charges on intraperitoneal photoimmunotherapy of ovarian cancer xenografts in nude mice. METHODS: The photosensitizer chlorin(e6) (c(e6)) was conjugated via a poly-l lysine linker to the F(ab')(2) fragment of the murine anti-ovarian cancer monoclonal antibody OC125, resulting in a photoimmunoconjugate with a pronounced cationic charge. Alternatively, by succinylating the poly-l-lysine conjugate, a photoimmunoconjugate with a pronounced anionic charge was obtained. A murine model of ovarian cancer derived from intraperitoneal inoculation of NIH:OVCAR-5 cells was employed. The conjugate was injected intraperitoneally followed after 3 h by red light delivered through a fiber into the peritoneal cavity. These photoimmunotherapy treatments were repeated three times, and the results obtained with the anionic and cationic photoimmunoconjugates were compared with those obtained with free c(e6) and control. The extent of residual macroscopic disease and death from disease were the evaluable outcomes for tumoricidal and survival studies, respectively. RESULTS: In contrast to other intraperitoneal photosensitizers, mice showed no systemic toxicity or morbidity from the treatment. In this initial study the mean residual tumor weights in all treatment groups ranged from 33 to 73 mg, as compared with 330 mg in untreated controls (P < 0.0001), and the response to the cationic conjugate was significantly better than that to the anionic conjugate or free c(e6) (P < 0.005). The median survival for mice treated with cationic photoimmunoconjugate was 41 days, compared with 35 days in controls (P = 0.009). CONCLUSION: Photoimmunotherapy with a cationic photoimmunoconjugate produces results superior to those obtained with an anionic conjugate, and further optimization of the treatment regimen may lead to a potential treatment for advanced ovarian cancer. PMID- 10684718 TI - Clinical applicability of the ATP cell viability assay as a predictor of chemoresponse in platinum-resistant epithelial ovarian cancer using nonsurgical tumor cell samples. AB - OBJECTIVES: There is no basis for choosing one chemotherapy over another in platinum-resistant epithelial ovarian cancer based on published response rates. This study explores the feasibility and accuracy of the ATP cell viability assay (ATP-CVA) in predicting chemoresponse in these difficult situations to choose the most appropriate drug for treatment. METHODS: Predominantly nonsurgical tumor samples for histological proof of recurrence were tested against a panel of drugs for salvage chemotherapy. Clinicians were blinded to the test results. Patient responses were evaluated after a minimum of three cycles of single-agent chemotherapy and correlated with test results. RESULTS: The evaluability rate was 85% (5 of 33 contaminated). The majority (24) were obtained by abdominal paracentesis and trucut biopsy. Of the 28 successful assays, 8 were excluded from analysis because four chose not to have chemotherapy and four withdrew after fewer than three cycles because of unacceptable side effects. The overall response rate to salvage chemotherapy was 15%. The sensitivity was 100% and specificity 82%. Resistance was correctly predicted in 100% and response correctly predicted in 50%. The outcomes of 17 of 20 patients were predicted correctly, giving an accuracy of 85%. CONCLUSIONS: It is feasible to test nonsurgical tumor specimens in recurrent cancer. The ATP-CVA correctly identified a group of patients with a 50% chance of response to salvage chemotherapy. This information may be useful in the decision-making process. A prospective, randomized study will be done to confirm these results. PMID- 10684719 TI - Unusual recurrence of cervical adenosquamous carcinoma after conservative surgery. AB - The use of less radical procedures for the treatment of early cervical cancers is gaining interest among physicians and young patients. Some authors have described surgical procedures aimed at reducing the surgical aggressiveness but the safety of such procedures remains debated. After a polypectomy, a young patient had a diagnosis of stage Ia(2) cervical adenosquamous carcinoma in 1995. As she wished to preserve her fertility, she underwent a cone biopsy and pelvic lymphadenectomy, without evidence of tumor spread. In 1998, at the 13th week of gestation, she had a diagnosis of a pelvic mass. The mass was a recurrence of carcinoma involving the myometrium, just underneath the peritoneum. She underwent a radical hysterectomy with bilateral oophorectomy. An ovarian metastasis was also detected at pathological exam. She received chemotherapy postoperatively and remains alive without evidence of disease. The recurrence of cervical cancer is traditionally regarded as an issue concerning the cervix, the parametria, or the lymph nodes. When the uterus is preserved we must also consider the possibility of a recurrence involving the corpus. With wider acceptance of limited therapeutic approaches we must be prepared for the detection of previously unknown patterns of recurrence and the follow-up modalities must be consequently adapted. PMID- 10684720 TI - Ovarian cancer in female-to-male transsexuals: report of two cases. AB - BACKGROUND: Ovarium cancer is the fifth most common cause of cancer-related death in women and is the most common fatal gynecologic malignancy. So far, ovarium carcinoma has not been reported to have occurred in female-to-male transsexuals. OBJECTIVE AND METHOD: We report on two such cases. Long-term exposure to increased levels of endogenous and exogenous androgens is hypothesized to constitute an additional risk factor in transsexuals as it has been associated with ovarian epithelian cancer. CONCLUSION: Simultaneous salpingo-oophorectomy should be performed in any female-to-male transsexual undergoing hysterectomy in the course of gender-confirming therapy. PMID- 10684721 TI - Hematogenous skin metastases from cervical cancer at primary presentation. AB - OBJECTIVE: Cutaneous metastasis from carcinoma of the uterine cervix is an uncommon occurrence. The majority of cases are diagnosed as recurrent carcinoma. This is believed to be the first report of hematogenous skin metastases present at the diagnosis of cervical carcinoma. METHODS: A case of a patient with cutaneous manifestations at the time of cervical carcinoma diagnosis is presented. RESULTS: Two lesions on the patient's hand occurred at points of recent skin puncture. These were biopsy-proven metastases from her primary cervical carcinoma. CONCLUSION: Skin metastases from cervical carcinoma are rare and represent a poor prognostic sign. PMID- 10684722 TI - Xp22.2-3 loss of heterozygosity is associated with germline BRCA1 mutation in ovarian cancer. AB - OBJECTIVE: X-Chromosome loss of heterozygosity (LOH) occurs in approximately 40% of ovarian cancers. We have previously demonstrated an association between nonrandom X-chromosome inactivation and germline BRCA1 mutation. The current study examines the association between X-chromosome LOH and BRCA1 mutation. METHODS: Ninety tumor DNA (81 ovary, 5 fallopian tube, 4 primary peritoneal) and matched peripheral blood mononuclear cell DNA samples were examined for LOH with 11 X-chromosome microsatellite DNA markers. RESULTS: Tumor DNA demonstrated frequent LOH at the Xp22.2-3 region (37.7% at DXS6807). Loss of heterozygosity on Xp was twice as common in tumor DNA from germline BRCA1 mutation carriers (9/14 vs 19/67, P = 0.02). In four evaluable samples, Xp22.2-3 LOH preferentially occurred from the active X allele. CONCLUSIONS: Our data support the hypothesis that an Xp22.2-3 gene product interacts with or modifies the expression of BRCA1 in some hereditary ovarian cancers. PMID- 10684723 TI - Sonographic follow-up in tamoxifene patients. PMID- 10684724 TI - Letters to the editor PMID- 10684725 TI - Restrictive endothelial barrier function during normal angiogenesis in vivo: partial dependence on tyrosine dephosphorylation of beta-catenin. AB - Differentiation of a restrictive endothelial barrier in the chick chorioallantoic membrane (CAM) occurs between Day 4.5 and Day 5.0 of the normal 21-day gestation. Whether molecular changes in the endothelial cell-cell junctional protein complex contribute to the ontogeny of barrier function represents the principal focus of this study. VE-cadherin has been shown to contribute to the regulation of endothelial cell monolayer permeability in vitro. Accordingly, VE-cadherin is complexed to the cytosolic catenins, and changes in monolayer permeability have been linked to alterations of the cadherin/catenin complex. Currently, a CAM endothelial VE-cadherin/beta-catenin complex was identified, and phosphotyrosine labeling of beta-catenin was decreased concurrently with the abrupt increase in CAM endothelial selectivity between Day 4.5 and Day 5.0. Further, inhibition of protein tyrosine phosphatases impeded regular tyrosine dephosphorylation of beta catenin at Day 5.0 and this served to partially restore macromolecular extravasation to elevated levels normally present at Day 4.5. Thus, differentiation of selective barrier function in the angiogenic CAM endothelium in vivo is dependent, in part, on tyrosine dephosphorylation of beta-catenin. PMID- 10684726 TI - Endothelin antagonists diminish postischemic microvascular incompetence and necrosis in the heart. AB - The endothelin receptor antagonists BQ-610 and BQ-123 were used to clarify the role of endothelin in the pathogenesis of postischemic microvascular incompetence in the myocardium. Forty-five isolated rat hearts were perfused with Krebs Henseleit buffer (KHB) for 15 min and then subjected to 0, 15, or 60 min of ischemia followed by 5 min of reperfusion with KHB, KHB + BQ-610, or KHB + BQ 123. They were fixed by perfusion with 2.5% glutaraldehyde and then perfused with nuclear track emulsion as an indicator of vascular flow. Transmural sections of resin-embedded myocardium were examined by scanning and transmission electron microscopy. Following 60 min of ischemia, the subendocardial third of the LV wall of hearts treated with BQ-123 showed nearly three times the proportion (P < 0.001) of competent capillaries in untreated hearts. Reperfusion after 15 min of ischemia of hearts treated with BQ-123 showed a 30% increase in the proportion of competent capillaries compared to controls (P < 0. 002). Treatment of corresponding groups with BQ-610 increased the proportion of competent capillaries but these differences were not statistically significant. In addition, both ET-I antagonists dramatically reduced the amount of ultrastructural change evident in myocardium reperfused after 60 min of ischemia. Thus endothelin plays a significant role in the pathogenesis of postischemic microvascular incompetence in the myocardium and, probably by its effects on Ca(2+) uptake, contributes also to the ultrastructural damage to the myocytes and endothelium which follows postischemic reperfusion of irreversibly injured myocardium. PMID- 10684727 TI - A novel model for the in vivo monitoring of uterine microcirculation and intracellular free calcium changes in rat. AB - The aim of this work was to develop a model to study the microcirculation and relative levels of intracellular free calcium in the myometrium of pregnant rats. On Day 21 of gestation a lobe of uterus was prepared free, flipped over, and mounted in a superfusion chamber leaving the radix and thereby the innervation and circulation intact. RBC velocity and arteriolar diameters were determined by means of intravital video microscopy before and after stimulation (norepinephrine). To study intracellular free calcium changes, the fluorescent dye Indo-1 AM was added to the superfusate in the chamber. Fluorescence images were recorded and ratios of the images collected at 400 and 506 nm were calculated and changes thereof were assumed to represent intracellular free calcium changes. RBC velocity and arteriolar diameter did not change for at least 1 h, while the response to norepinephrine was similar at the beginning of the experiment and after 120 min. In four separate interventions, the uterus was challenged with 5 x 10(-4) IU/ml oxytocin, 4.5 mM calcium, 5 x 10(-4) IU/ml oxytocin with 4.5 mM calcium, and 5 microM ionomycin, resulting in an increase of the 400/506 nm ratio of 27, 31, 76, and 103%, respectively, representing a relative increase in intracellular free calcium. This novel in vivo model is suitable for monitoring intracellular free calcium changes and to record RBC velocities and blood vessel diameters in the myometrium of pregnant rats. PMID- 10684728 TI - Generational analysis reveals that TGF-beta1 inhibits the rate of angiogenesis in vivo by selective decrease in the number of new vessels. AB - Quantitative analysis of vascular generational branching demonstrated that transforming growth factor-beta1 (TGF-beta1), a multifunctional cytokine and angiogenic regulator, strongly inhibited angiogenesis in the arterial tree of the developing quail chorioallantoic membrane (CAM) by inhibition of the normal increase in the number of new, small vessels. The cytokine was applied uniformly in solution at embryonic day 7 (E7) to the CAMs of quail embryos cultured in petri dishes. After 24 h the rate of arterial growth was inhibited by as much as 105% as a function of increasing TGF-beta1 concentration. Inhibition of the rate of angiogenesis in the arterial tree by TGF-beta1 relative to controls was measured in digital images by three well-correlated, computerized methods. The first computerized method, direct measurement by the computer code VESGEN of vascular morphological parameters according to branching generations G(1) through G(>/=5), revealed that TGF-beta1 selectively inhibited the increase in the number density of small vessels, N(v>/=5) (382 +/- 85 cm(-2) for specimens treated with 1 microg TGF-beta1/CAM for 24 h, compared to 583 +/- 99 cm(-2) for controls), but did not significantly affect other parameters such as average vessel length or vessel diameter. The second and third methods, the fractal dimension (D(f)) and grid intersection (rho(v)), are statistical descriptors of spatial pattern and density. According to D(f) and rho(v), arterial density increased in control specimens from 1.382 +/- 0.007 and 662 +/- 52 cm(-2) at E7 (0 h) to 1.439 +/- 0.013 and 884 +/- 55 cm(-2) at E8 (24 h), compared to 1. 379 +/- 0.039 and 650 +/ 111 cm(-2) for specimens treated with 1 microg TGF-beta1/CAM for 24 h. TGF-beta1 therefore regulates vascular pattern and the rate of angiogenesis in a unique "fingerprint" manner, as do other major angiogenic regulators that include VEGF, FGF-2 (bFGF), and angiostatin. TGF-beta1 did not stimulate angiogenesis significantly at low cytokine concentrations, which suggests that this quail CAM model of angiogenesis is not associated with an inflammatory response. PMID- 10684729 TI - Heterogeneity in cytosolic calcium regulation among different microvascular smooth muscle cells of the rat retina. AB - Rat retinae were dissociated to yield intact microvessels 7 to 42 microm in diameter. These were loaded with fura-2 AM and single fragments anchored down in a recording bath. Intracellular Ca(2+) levels from 20- to 30-microm sections of vessel were estimated by microfluorimetry. The vessels studied were identified as metarterioles and arterioles. Only the microvascular smooth muscle cells loaded with fura-2 AM and changes in the fluorescence signal were confined to these cells: Endothelial cells did not make any contribution to the fluorescence signal nor did they contribute to the actions of the drugs. Caffeine (10 mM) or elevated K(+) (100 mM) produced a transient rise in cell Ca(2+) in the larger vessels (diameters >18 microm) but had no effect on smaller vessels (diameters <18 microm). Rises in cell Ca(2+) were accompanied by a rapid ( approximately 2 s to peak) contraction followed by relaxation. Caffeine and K(+) responses were blocked by ryanodine (10 microM) and nifedipine (1 microM), respectively. In all the vessels tested, vasopressin (arginine, 10 nM) elicited a transient increase in cell Ca(2+) and a constriction, irrespective of the diameter of the vessel. All vessels tested also responded to endothelin-1 (1-10 nM) through an Et(A) receptor to produce a transient rise in cell Ca(2+) followed by a plateau phase of elevated Ca(2+) and a constriction. In contrast to the transient effects of vasopressin, caffeine, and K(+), the cell Ca(2+) remained elevated (>30 min) on washing out the endothelin and the vessel failed to relax. These results demonstrate heterogeneity between smaller and larger retinal vessels with regard to Ca(2+) mobilisation and homogeneity with respect to the actions of vasoactive peptides. PMID- 10684730 TI - The effect of treatment with low dose ACE inhibitor and/or diuretic on coronary microvasculature in stroke-prone spontaneously hypertensive rats. AB - Angiotensin II is considered to have angiogenic properties. Nevertheless, several authors reported an increase in coronary capillary density after treatment with ACE inhibitors. The aim of the present study was to evaluate the effect of treatment with low doses of ACE inhibitor perindopril, low doses of the diuretic indapamide, or a combination of the two on microvascular structure in hearts from stroke-prone spontaneously hypertensive rats (SHR-sp). Young adult male SHR treated with indapamide (0.24 mg/kg/day), perindopril (0.76 mg/kg/day), or both were compared with untreated animals after 8 or 14 weeks of treatment. Survival of SHR-sp was significantly increased after treatment. Only perindopril alone or in combination with indapamide significantly decreased blood pressure and cardiac mass. Treatment also significantly increased capillary and myocyte densities but arteriolar density tended to decrease. External and internal diameters significantly increased in treated animals while arteriolar thickness remained the same. Thus, thickness in vessels of the same size was the greatest in untreated animals, followed by indapamide- and perindopril-treated rats with the thinnest walls in rats with combined treatment, and the treatment resulted in a significant increase in the lumen to wall ratio. Capillary and arteriolar growth responses in treated animals seem to indicate that the two are independently regulated processes. Treatment with indapamide alone at this dosage did not significantly influence most responses but in combination with perindopril it strengthened the effect of perindopril. PMID- 10684731 TI - Reduced renal mass hypertension, but not high salt diet, alters skeletal muscle arteriolar distensibility and myogenic responses. AB - The effects of high salt diet and reduced renal mass hypertension (RRM-HT) on skeletal muscle arteriolar distensibility and myogenic responses were investigated in male Sprague-Dawley rats. Rats were enclosed in an air-tight box with the in situ cremaster muscle exteriorized and viewed via television microscopy. Normotensive rats were fed low salt (0.4% NaCl) or high salt (4.0% NaCl) diet and RRM-HT rats were fed high salt diet for 4-6 weeks. With the cremaster muscle superfused with either physiological salt solution (for myogenic responses) or Ca(2+)-free physiological salt solution (for arteriolar distensibility), box pressure (and therefore, intravascular pressure) was increased in 5 mm Hg increments to a maximum of +30 mm Hg. The myogenic responses of arterioles were not altered by high salt diet, but were enhanced with RRM-HT. Arteriolar distensibility was not affected by high salt diet, but was reduced in RRM-HT rats compared to either normotensive rat group. These data suggest that high salt diet does not significantly alter either myogenic responses or the distensibility of rat cremasteric arterioles. However, RRM-HT enhances myogenic responses of these vessels while reducing arteriolar distensibility. The impact of these effects must be taken into account when interpreting data describing alterations in skeletal muscle microvessel reactivity for animals on high salt diet or with RRM hypertension. PMID- 10684732 TI - Effect of vascular endothelial growth factor on cultured endothelial cell monolayer transport properties. AB - Vascular endothelial growth factor (VEGF) is a potent enhancer of microvascular permeability in vivo. To date, its effects on hydraulic conductivity (L(p)) and diffusive albumin permeability (P(e)) of endothelial monolayers have not been thoroughly assessed in vitro. We hypothesized that VEGF affects endothelial transport properties differently depending on vessel location and endothelial phenotype. Using three well-established endothelial cell culture models-human umbilical vein endothelial cells (HUVECs), bovine aortic endothelial cells (BAECs), and bovine retinal microvascular cells (BRECs)-grown on porous, polycarbonate filters we were able to produce baseline transport properties characteristic of restrictive barriers. Our results show 3.1-fold and 5.7-fold increases in endothelial L(p) for BAEC and BREC monolayers, respectively, at the end of 3 h of VEGF (100 ng/ml) exposure. HUVECs, however, showed no significant alteration in L(p) after 3 h (100 ng/ml) or 24 h (25 ng/ml) of incubation with VEGF even though they were responsive to the inflammatory mediators, thrombin (1 U/ml; 27-fold increase in L(p) in 25 min) and bradykinin (10 microM; 4-fold increase in L(p) in 20 min). Protein kinase C (PKC) and nitric oxide (NO) are downstream effectors of VEGF signaling. BAEC L(p) was responsive to activation of NO (SNAP) and PKC (PMA), whereas these agents had no effect in altering HUVEC L(p). Moreover, BAECs exposed to the PKC inhibitor, staurosporine (50 ng/ml), exhibited significant attenuation of VEGF-induced increase in L(p), but inhibition of nitric oxide synthase (NOS) with L-NMMA (100 microM) had no effect in altering the VEGF-induced increase in L(p). These data provide strong evidence that in BAECs, the VEGF-induced increase in L(p) is mediated by a PKC-dependent mechanism. Regarding diffusive albumin P(e), at the end of 3 h, BAECs and BRECs showed 6.0-fold and 9. 9-fold increases in P(e) in response to VEGF (100 ng/ml), whereas VEGF had no significant effect after 3 h (100 ng/ml) or 24 h (25 ng/ml) in changing HUVEC P(e). In summary, these data indicate that VEGF affects endothelial transport properties differently depending on the vessel type and that differences in cell signaling pathways underlie the differences in VEGF responsiveness. PMID- 10684733 TI - Assessment of angiogenesis and tumor growth in conscious mice by a fluorimetric method. AB - Angiogenesis and tumor growth in conscious mice have been determined using the kinetics of appearance of a fluorescent tracer in the bloodstream after application to subcutaneously implanted sponges bearing tumor cells. The functional parameter expressed in terms of half-time (t(1/2); time taken for the fluorescence to reach 50% of the peak in the systemic circulation), which is inversely proportional to blood flow, showed that in the tumor-free implants t(1/2) values decreased from 11.55 +/- 1 min at day 1 to 5.7 +/- 0. 44 min by day 14. In the tumor-bearing implants, this process was accelerated and maximum vascularization was achieved by day 7 (3 days after tumor cell inoculation). Increases in t(1/2) values were observed at days 10 and 14, which paralleled the tumor growth as indicated by wet weight. The hemoglobin content (microg Hb/mg wet weight) in the tumor-free group increased during the 14-day period. In contrast, in the tumor-bearing implants. Hb concentration decreased per unit of tissue weight. Dexamethasone treatment for 13 days prevented fibrovascular tissue infiltration in tumor-free implants, but was unable to delay tumor growth, indicating that this procedure can be used to exclude the inflammatory reaction induced by the implantation technique, thus allowing tumor angiogenesis to be studied without the confounding influence of the host inflammatory cells. The results of our experimental observation indicate the suitability of this combination of techniques for analyzing angiogenesis induced by tumor cells and several hemodynamic features of Ehrlich tumor growth in awake animals. PMID- 10684734 TI - Vascular endothelial growth factor regulates both apoptosis and angiogenesis of choriocapillaris endothelial cells. PMID- 10684735 TI - A histomorphometric observation of flows in cortical bone under dynamic loading. AB - Ferritin was used as a histological marker for interstitial fluid flows in four goats. Two transcortical pins were inserted into each tibia mediolaterally-one at the proximal side and one at the distal side of the diaphysis. For the experimental limb, dynamic loading was applied to the pins on the lateral side. The contralateral limb was used as control. Ferritin was injected into the nutrient arteries feeding the two tibiae. The goat was then euthanized immediately. Undecalcified slides of the tibial cortical bone from both the experimental and the control limbs were studied histologically. Percentages of Haversian systems marked with ferritin halos and the average radial distance of ferritin transudation away from the Haversian canals were assessed. Results for the medial and lateral sides of both the experimental and control tibiae were reported. Significant differences in the ferritin transudation distance were found between the experimental and the control tibia (P < 0.005) and between the medial and the lateral sectors (P < 0.05). The approach documented in this paper could be used to address systematically how external loading might affect the transport phenomena in cortical bone. PMID- 10684736 TI - Assessment of continuous skeletal muscle blood flow during exercise in humans. AB - The ability to measure regional blood flow from exercising skeletal muscles is of great interest. However, noninvasive techniques such as venous occlusion plethysmography and pulsed Doppler duplex ultrasonography only allow determination of blood flow at rest. The aim of our study was to investigate the influence of position on continuous measured skeletal muscle blood flow response in the upright and supine positions during graded maximal exercise by means of the local (133)Xenon washout technique with portable CdTe(Cl) detectors. Fifteen healthy subjects (8 women and 7 men, mean age 46 +/- 11 years) performed graded maximal bicycle exercise in both supine and upright positions in random order on 2 subsequent days. Blood flow in the musculus tibialis anterior was measured using the local (133)Xenon washout technique. A total of 55-110 MBq of (133)Xenon dissolved in isotonic saline was injected intramuscularly and the gamma emission was registered by light-weight portable CdTe(Cl) detectors. During supine exercise skeletal muscle blood flow increased continuously with increasing work load. However, during upright exercise blood flow increased only at the initial three work loads, then it decreased gradually. Immediately after exercise blood flow returned to preexercise values for both positions. The skeletal muscle blood flow at maximum work load for each subject was 74% (relative flow values) (P < 0.05) higher in the supine compared with the upright position. There was no significant difference in absolute or relative blood flow values at similar time points. Exercise time was longer in the supine (1345 +/- 548 s) compared with the upright position (1148 +/- 453 s) (P < 0.005). The local (133)Xenon washout technique with portable CdTe(Cl) detectors allows continuous determination of skeletal muscle blood flow during graded bicycle exercise in supine and upright positions. Furthermore, blood flow at maximum work load and exercise time was increased in supine compared with upright exercise. PMID- 10684737 TI - Patterns of conducted vasomotor response in the mouse. PMID- 10684738 TI - Erythrocytes enhance lymphocyte rolling and arrest in vivo. PMID- 10684739 TI - Altered structure and mechanics of skeletal muscle arteries with high-salt diet and reduced renal mass hypertension. PMID- 10684740 TI - Child pedestrian injury prevention project: student results. AB - BACKGROUND: Few comprehensive pedestrian safety interventions for primary-school age children have been developed and evaluated. This paper reports the impact of the 3-year (1995-1997) Child Pedestrian In jury Prevention Project (CPIPP) on a cohort of 1603 children followed from age 6 to 9 years. This multicomponent project comprised an educational intervention for students, their parents and teachers, and the local community, as well as several environmental interventions. The primary aim of CPIPP was to improve children's road-related behavior and to enhance the safety of their road environment. METHODS: Three communities were assigned to the treatment conditions: (1) high-education, community, and environmental interventions; (2) moderate-education intervention only; and (3) comparison (usual road safety education). Children's pedestrian knowledge and road crossing and playing behaviors were assessed using a pre- and posttest self-report questionnaire. Their self-reported road crossing behaviors were validated using an observational schedule and brief interview. RESULTS: Children in the high and moderate intervention groups were significantly more likely to cross the road with adult supervision (P = 0.013) and play away from the road (P = 0.000) than the comparison group. No differences were detected in children's pedestrian safety knowledge between the intervention and comparison groups. CONCLUSIONS: While several methodological limitations may have influenced the study outcomes, these data nonetheless indicate that in the study sample the CPIPP educational intervention deaccelerated the natural increase in children's pedestrian-related risk behavior. PMID- 10684741 TI - Carpal tunnel syndrome. A message from preventive medicine and your physician PMID- 10684742 TI - Leisure-time, occupational, and household physical activity among professional, skilled, and less-skilled workers and homemakers. AB - BACKGROUND: Adults in lower status occupations are at higher risk of premature cardiovascular disease, for which physical inactivity is a major risk factor. While lower rates of leisure-time physical activity have been found to be associated consistently with lower income and education levels, the association between occupational and home-based physical activity with membership of different occupational categories is not well understood. METHODS: An urban representative population data set derived from a self-completion questionnaire was used to examine both self-reported leisure-time physical activity and a combined measure of occupational/home-based physical activity of adult less skilled, skilled, and professional workers and homemakers (3795 males; 4140 females). chi(2) analyses, ANOVA, and logistic regressions were used to examine the associations between occupational group membership and physical activity. RESULTS: After adjustment for age, body mass index, education, country of birth, marital status, and smoking, less-skilled workers were less likely to report any form of leisure-time physical activity. However, occupational category was not a strong predictor of participation in combined vigorous occupational/home physical activity. Homemakers and those in lower status occupations were less likely to report participation in vigorous leisure-time physical activity sufficient for cardiorespiratory fitness. With the inclusion of time spent in combined vigorous occupational/home physical activity, there was no longer an association of activity with occupational status for males. However, for females the association remained. CONCLUSIONS: The assessment of occupational and household physical activity in addition to leisure-time activity may be important for understanding associations between occupational categories, physical activity, and increased levels of health risk and for the development of physical activity promotion strategies. PMID- 10684743 TI - A cross-cultural consumer-based decision aid for screening mammography. AB - BACKGROUND: "Women should have mammograms" is the usual perspective of educational interventions about screening. The perspective that "women should be informed" about potential risks and benefits so they can make value- and evidence integrated personal decisions has recently been advocated. However, this perspective has not previously been operationalized. We developed an evidence based cross-cultural mammography decision aid (MDA), for European American and Mexican American women who are 50-70 years old, at average risk of breast cancer, of varying educational levels, and English- or Spanish-speaking. METHODS: MDA development included: (1) content development by a multidisciplinary team and lay women and (2) testing for validity and reliability. Four parts include: (1) introduction; (2) information about logistics (cost, time, discomfort) and risks (sequelae of false-positive or negative results; (3) probability of developing breast cancer; and (4) benefit of mammography regarding breast cancer outcomes (e.g., death and recurrence). We assessed reliability (stability of decisions with the same information) after 1-2 weeks. We assessed validity (comprehension of information) quantitatively (probabilities were changed to see whether preferences changed predictably) and qualitatively (focus groups, standardized probes for comprehension). Subjects were a convenience sample of 49 European American (50-81 years old) and 54 Mexican American (49-89 years old) women from administrative staff at a medical school, the waiting room of an indigent primary care clinic, and a community center. RESULTS: Reliability was 100%. In quantitative validity testing, 22 of 28 women (89%) changed preference as predicted with changed probabilities. Comprehension was confirmed qualitatively in all phases of testing with both Spanish and English versions. CONCLUSION: The decision aid is valid and reliable in English and Spanish for southwestern Mexican American and European American women at average risk of breast cancer, including those of low educational levels. PMID- 10684744 TI - Cost-benefit analysis of sustained-release bupropion, nicotine patch, or both for smoking cessation. AB - BACKGROUND: The nicotine transdermal patch (NTP) has been shown previously to be a cost-effective smoking cessation intervention. This is the first economic analysis comparing the NTP with the only non-nicotine-containing pharmacological intervention, bupropion HCl. METHODS: Decision-tree analysis, based on a previously published cost-benefit smoking-cessation model, was used to determine the optimal treatment from the standpoint of costs versus benefits, from the employer's perspective. Base-case probabilities of successful quitting in our model came from clinical trial point-prevalence data at the end of a 1-year follow-up study (N = 893) comparing placebo, bupropion, NTP, and bupropion/NTP in combination, administered along with minimal counseling. Sensitivity analyses were performed to determine the effects of variations in base-case assumptions regarding the monetary benefits that would accrue if an intervention were successful, probabilities of quitting, drug costs, cost of lost work time for a health care provider visit, and cost of the visit itself. RESULTS: The analysis showed that bupropion is more cost-beneficial than either NTP or bupropion/NTP, with a net benefit in the first post-quit year of up to $338 per employee who attempts to quit compared with $26 for NTP, $178 for the two in combination, and $258 for placebo. These results were robust to most plausible variations in the assumptions used in the model. One exception was the monetary benefit of successful intervention (assumed in the base-case to be $1,654). If this benefit were actually less than $1, 112, placebo (i.e., minimal counseling with no pharmacological intervention) would be more cost-beneficial than any of the active treatments. CONCLUSION: From an employer's perspective, bupropion 300 mg/day for 9 weeks is a more cost-beneficial smoking cessation intervention than the nicotine patch, and under most scenarios, bupropion is also more cost beneficial than placebo. PMID- 10684745 TI - Associations between exercise and health behaviors in a community sample of working adults. AB - BACKGROUND: The present study examined the associations between leisure-time exercise and a range of health behaviors and reports of illness and injury in a sample of community working adults. METHODS: The study population included 4907 women and 4136 men who completed surveys in 24 worksites in the Minneapolis-St. Paul metropolitan area. RESULTS: Participants in the study were ranked by gender according to their exercise score and grouped into quartiles. Women and men in the highest activity quartiles were more highly educated and were younger. High activity men were more likely to be unmarried. Higher levels of leisure-time exercise were positively associated with seat belt use and inversely related to smoking, dietary fat intake, reported stress, and obesity. In men only, leisure time exercise was related to greater reports of injuries resulting in restriction of usual activities. In women only, leisure-time exercise was positively associated with daily alcohol use. Most of the significant associations were seen in the two highest quartiles of exercise. CONCLUSIONS: These findings suggest that associations between leisure-time exercise and health behaviors occur at the higher levels of exercise and interventions may need to promote this higher level of leisure-time exercise to impact overall public health. PMID- 10684746 TI - Vegan diet-based lifestyle program rapidly lowers homocysteine levels. AB - BACKGROUND: Plasma homocysteine levels have been directly associated with cardiac disease risk. Current research raises concerns as to whether comprehensive lifestyle approaches including a plant-based diet may interact with other known modulators of homocysteine levels. METHODS: We report our observations of homocysteine levels in 40 self-selected subjects who participated in a vegan diet based lifestyle program. Each subject attended a residential lifestyle change program at the Lifestyle Center of America in Sulphur, Oklahoma and had fasting plasma total homocysteine measured on enrollment and then after 1 week of lifestyle intervention. The intervention included a vegan diet, moderate physical exercise, stress management and spirituality enhancement sessions, group support, and exclusion of tobacco, alcohol, and caffeine. B vitamin supplements known to reduce blood homocysteine levels were not provided. RESULTS: Subjects' mean homocysteine levels fell 13%: from 8.66 micromol/L (SD 2.7 micromol/L) to 7.53 micromol/L (SD 2.12 micromol/L; P < 0.0001). Subgroup analysis showed that homocysteine decreased across a range of demographic and diagnostic categories. Conclusions. Our results suggest that broad-based lifestyle interventions favorably impact homocysteine levels. Furthermore, analysis of Lifestyle Center of America program components suggests that other factors in addition to B vitamin intake may be involved in the observed homocysteine lowering. PMID- 10684747 TI - Black/white differences in relative weight and obesity among girls: the Bogalusa Heart Study. AB - BACKGROUND AND OBJECTIVE: Although black women have a higher prevalence of overweight and obesity than do white women, it is unclear if a similar pattern exists among youths. We therefore examined the development of black/white differences in relative weight and adiposity among 5 to 17-year-old girls. METHODS: Cross-sectional analyses of 4542 black and 4542 white girls who were examined between 1973 and 1994. Quetelet Index (kg/m(2)), Rohrer Index (kg/m;s(3)), and height-adjusted weight were used as measures of relative weight, and subscapular and triceps skinfolds as measures of adiposity. Breast development was used as an index of sexual maturation. RESULTS: On average, black girls were 1 to 3 kg heavier than were similarly aged white girls, and before adolescence, they were 2 to 3 cm taller. After adjusting for differences in height, the mean relative weight of black girls was consistently greater than that of white girls only after age 13; furthermore, sexual maturation was a stronger correlate of relative weight among black girls than among white girls. Comparable differences were seen for the subscapular skinfold thickness, but white girls consistently had a thicker mean triceps skinfold than did black girls. CONCLUSION: Sexual maturation should be considered in comparisons of relative weight and obesity among youths, and as compared with white girls, black girls do not have a higher mean relative weight until adolescence. The use of different indices of overweight and adiposity can lead to contrasting results, with simple comparisons of Quetelet Index tending to overstate the relative weights of taller children. PMID- 10684748 TI - A randomized controlled trial of a clinic-based support staff intervention to increase the rate of fecal occult blood test ordering. AB - BACKGROUND: Colorectal cancer is the second most common fatal malignancy in the United States. Early detection using fecal occult blood tests has been shown to reduce mortality, but these tests are underutilized among those eligible for this screening. Attempts to increase use of fecal occult blood tests in eligible populations have focused on the provider, patient, or system. But none have examined whether a support-staff intervention is effective in achieving this aim. We therefore conducted a randomized controlled trial to test the impact of authorizing support staff to order fecal occult blood tests in a general internal medicine clinic organized into four teams. METHODS: A total of 1,109 patients were included in the study, 545 of whom were in the two teams randomized to treatment. Univariate and multivariate regression analyses were used to evaluate the impact of the intervention. RESULTS: The intervention resulted in significantly more fecal occult blood test ordering in the treatment group than in the control group for all patients (52% vs 15%, P < 0.001). Treatment fecal occult blood test cards were returned as frequently as the control cards for all patients (44% vs 48%, P = 0.571). CONCLUSION: Delegation of selected screening tasks to support staff can enhance patient access to preventive care. PMID- 10684750 TI - Publishers announcement. PMID- 10684749 TI - Trends in plasma cholesterol levels in the atherosclerosis risk in communities (ARIC) study. AB - BACKGROUND: Data from the Atherosclerosis Risk in Communities (ARIC) cohort study were examined both cross-sectionally and intraindividually to confirm recent findings from population-based studies showing a decline in total cholesterol (TC) levels in the United States. METHODS: For the cross-sectional analysis, mean plasma TC levels from 15,792 participants aged 45-64 at baseline visit, and who were selected randomly from four U.S. communities, were examined for each year covered by the first cohort visit (1987, 1988, and 1989). Ninety-three percent of the cohort participants returned for the follow-up visit (1990, 1991, and 1992), and were included in the assessment of intraindividual TC trends. RESULTS: Both mean TC and prevalence of hypercholesterolemia (defined as plasma cholesterol concentration >/=240 mg/dl) consistently declined over the 3 years covered by visit 1 for all age-gender-race groups. For 1987, 1988, and 1989, mean TC values (mg/dl) were, respectively, 220.3, 216.7, and 214.1 (annual average change, 1.4%, P < 0.001). For these same years, hypercholesterolemia prevalence rates were 30. 0, 27.8, and 25.3% (annual average change, -7.8%, P < 0.001). The mean plasma TC also decreased within individuals between the two visits across race, gender, and age decade categories. With the exception of black men, this decline was more marked for older than younger subjects, but no consistent differences were seen between the racial groups. However, in whites, decreases were greater for men than for women. Expected results were seen when these changes were correlated with changes in cardiovascular risk factors between the two visits. CONCLUSION: The current study results are consistent with those of previous studies, and confirm the notion that preventive programs appear to be effective in reducing mean population TC levels. PMID- 10684751 TI - Sialography of sheep parotid and mandibular salivary glands. AB - The anatomy of ovine salivary glands was studied on cadaver heads. The mandibular duct enters the oral cavity on the ventral surface of sublingual caruncles, which are located medial to the orifice of the ventral sublingual gland duct. The parotid gland duct enters the oral cavity on the cheek opposite the upper 2nd molar. Prior to applying sialography to live animals, the procedure was carried out on cadaver heads then the live animals were sedated, the mandibular and parotid ducts catheterized and contrast medium was injected into each gland. Lateral radiographs were made immediately after the injection. The normal sheep mandibular and parotid salivary glands have a multilobular appearance in cadaver heads, but in live animal only the ducts and their smaller branches could be identified. The mean diameter of mandibular and parotid duct were 1.4+/-0.3 mm and 3. 1+/-1.0 mm respectively. The monostomatic sublingular gland had a slender shape in the sialogram. In conclusion sialography of mandibular, parotid and sublingual salivary glands in sheep is practical and can be helpful in diagnosis of pathological conditions of these glands. PMID- 10684752 TI - Simultaneous serological evidence of Actinobacillus pleuropneumoniae, PRRS, Aujeszky's disease and influenza viruses in Spanish finishing pigs. AB - A total of 198 pigs with tachypnoea and temperature >/= 40 degrees C were selected on a Spanish finishing unit, and their sera were examined for antibodies to Actinobacillus pleuropneumoniae (App), porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky' disease virus (ADV), and swine influenza virus (SIV). Eighty-nine point nine per cent of the pigs were seropositive to App, 88.6 per cent to PRRS, 73.0 per cent to ADV, and 30.6 per cent to SIV. Thirty-one pigs (15.6 per cent) were seropositive for App, PRRSV, ADV and SIV, and only one (0.5 per cent) was seronegative for all. Statistical association was assessed for dual infections but it was not found in any case (P > 0.05). Other parameters (dyspnoea, nasal discharge and coughing) were also recorded, and no significant associations between them and the presence of antibodies against any of the four infections was found. PMID- 10684754 TI - Detection of Lawsonia intracellularis in the tonsils of pigs with proliferative enteropathy. AB - The presence of Lawsonia intracellularis, the obligate intracellular bacterium causing proliferative enteropathy (PE), in the tonsils of pigs as a locus for infection or extraintestinal occurrence of the bacterium was investigated by PCR and immunohistochemistry. Tonsillar occurrence of L. intracellularis could be part of the pathogenesis of PE and an important risk factor in the spread of the disease. L. intracellularis was detected by only PCR in the tonsils of 2/32 pigs without PE at necropsy but with a clinical history of diarrhoea and detection of the bacterium in faeces 1 to 3 weeks prior to necropsy but not in four pigs with moderate PE lesions. However, L. intracellularis was detected in the tonsils of 4/9 pigs with PE complicated with necroses and in 4/4 pigs with proliferative haemorrhagic enteropathy in which L. intracellularis antigen also was demonstrated in tonsillar macrophages and as intact bacteria in the lumen of the crypts. The results show that L. intracellularis is detectable in the tonsils of pigs and that the tonsillar presence of L. intracellularis appears to be correlated to the severity of the intestinal lesions possibly as a result of local retention and not as part of the pathogenesis of PE. PMID- 10684753 TI - Oestrogen and progesterone receptors in feline fibroadenomatous change: an immunohistochemical study. AB - The distribution of oestrogen and progesterone receptors was analysed in 18 cases of feline fibroadenomatous change (FFAC) using commercially available specific monoclonal antibodies and the Avidin-biotin peroxidase complex technique on formalin-fixed, paraffin-embedded tissue. In all cases of FFAC, progesterone receptors were detected either in epithelial cells (mostly suprabasal) or in epithelial and stromal cells. Oestrogen receptors were detected in approximately half of these cases, in suprabasal or luminal epithelial cells exclusively. Myoepithelial cells lacked both oestrogen and progesterone receptors. The techniques used have identified the specific cellular distribution of steroid hormones receptors in this hormone-dependent lesion of the feline mammary gland. The results confirm those of previous biochemical analyses with respect to progesterone receptors and add new data concerning the possible involvement of oestrogen receptors and stromal fibroblasts in the hormonal control and development of the lesion. PMID- 10684755 TI - Optimal conditions for simultaneous measurement of platelet aggregation and ATP secretion in canine whole blood. AB - This paper describes the optimal conditions for simultaneous evaluation of platelet aggregation and secretion capacity in canine whole blood using a Whole Blood Lumi-Aggregometer (Chrono-Log Corporation, Havertown, Pensylvania). For this purpose, the potential influence of several parameters was investigated using collagen, adenosinediphosphate (ADP), arachidonic acid (AA) and thrombin as platelet agonists. Results indicate that optimal experimental conditions to obtain reliable results include: allowing blood samples to stand at room temperature 60 minutes after blood collection, analysing samples within 3 hours from time of collection, adjusting platelet numbers to a final concentration of 150 000 microl(-1)and mixing the sample with isotonic saline (1:1) before adding the platelet agonist. The use of different platelet agonists offers variable results: collagen (0.5, 1 and 5 microg ml(-1)) is suitable for simultaneous platelet aggregation and adenosintriphosphate (ATP) secretion measurements; 1 UI ml(-1)of thrombin induced maximum ATP secretion;AA (0.5 and 1 mM) and ADP (5, 10 and 25 microM) did not give consistent results. The method described in this study has important clinical applications since it allows easy and quick platelet function evaluation in pathologic states. PMID- 10684756 TI - Determination of intraspecies variations of the V2 region of the 16S rRNA gene of Streptococcus equi subsp. zooepidemicus. AB - The 16S rRNA gene of 39 S. equi subsp. zooepidemicus strains and two S. equi subsp. equi strains was amplified by polymerase chain reaction and subsequently digested with the restriction enzyme Hinc II. A restriction profile with two fragments with sizes of 1250 bp and 200 bp could be observed for both S. equi subsp. equi strains and for 30 of the 39 S. equi subsp. zooepidemicus strains indicating a sequence variation within the V2 region of the 16S rRNA gene of the remaining nine S. equi subsp. zooepidemicus isolates. A segment of the 16S rRNA gene including the hypervariable V2 region of 11 S. equi subsp. zooepidemicus and two S. equi subsp. equi could be amplified by PCR and sequenced. The sequence of the V2 region of eight S. equi subsp. zooepidemicus strains appeared to be identical or almost identical to the sequence of the two S. equi subsp. equi strains. The sequence of the remaining three S equi subsp. zooepidemicus strains differed significantly from the sequence of S. equi subsp. equi. These differences allowed a division of S. equi subsp. zooepidemicus strains into two 16S rRNA types and might possibly have consequences for the taxonomic position of these phenotypically indistinguishable strains of one subspecies. A molecular typing could additionally be performed by amplification of the gene encoding the 16S-23S rRNA spacer region. A single amplicon of the spacer gene of 1100 bp could be observed for one S. equi subsp. zooepidemicus, an amplicon of 950 bp for two S. equi subsp. equi strains and 10 S. equi subsp. zooepidemicus strains, a amplicon of 780 bp for 27 S. equi subsp. zooepidemicus strains and a single amplicon of 600 bp for one S. equi subsp. zooepidemicus strain. The variations of the V2 region of the 16S rRNA gene and the size variations of the 16S-23S rRNA spacer gene were not related to each other. Both variations could be used for molecular typing of this species, possibly useful in epidemiological aspects. PMID- 10684757 TI - An interobserver and intraobserver study of buccal mucosal bleeding time in Greyhounds. AB - Two observers experienced with the buccal mucosal bleeding-time technique using a standardised device (Surgicutt) performed the test on 20 Greyhounds, to evaluate interobserver and intraobserver repeatability. The interobserver and intraobserver repeatability were both about 2 minutes. The results indicated that, for any two readings within a dog, the buccal mucosal bleeding time may differ by up to +/- 2 minutes. A single reading was accurate to within +/- 80 seconds. Sixty-one Greyhounds were used to establish a reference interval for the buccal mucosal bleeding time, and to assess the relationship between the buccal mucosal bleeding time and plasma von Willebrand factor concentration. The mean was 129.5 (SD 44.2) seconds. The reference interval was 53 to 235 seconds, which was slightly lower than non-greyhounds. No significant correlation (r=-0. 18, P=0.17) between the buccal mucosal bleeding time and plasma von Willebrand factor concentration was found in the 61 Greyhounds, where plasma von Willebrand factor concentration was in the range 29 to 160 Canine Units dL(-1). PMID- 10684758 TI - Mechanics of the respiratory system in healthy newborn calves using impulse oscillometry. AB - Arterial blood gases, acid-base balance and respiratory function tests using impulse oscillometry (IOS) were performed on 40 clinically healthy newborn calves during the first 24 hours of life to evaluate their respiratory adaptation to extrauterine life. Gas exchange efficiency of the lung was significantly improved with time and was accompanied by the correction of the mixed acidosis observed at birth and by significant changes in respiratory mechanics. Major changes were detected within the first 6 hours. The significant decrease in resistance (R) and the increase in reactance (X) with time, demonstrate the improvement in respiratory mechanics of both upper and lower airways, and reflect the increase in lung volume, the improved lung tissue elasticity and/or distribution of the ventilation. Respiratory mechanical, arterial blood gases and acid-base balance data provided in this study describe a successful respiratory adaptation to extrauterine life in healthy newborn calves. PMID- 10684759 TI - Differentiation induction of canine osteosarcoma cell lines by retinoids. AB - The effect of two retinoids, all- trans and 9- cis retinoic acid, on the differentiation of three canine osteosarcoma cells (OOS, HOS, and POS) was examined using markers specifically expressed by phenotypic osteoblasts. Both retinoids induced morphologic differentiation in all the canine osteosarcoma cells. Retinoids enhanced cell flattening and spreading, as well as reduction in cell overlapping. Alkaline phosphatase (ALP) activity and ALP staining was enhanced in OOS, and HOS cells, but decreased in POS cells. These results may suggest that OOS and HOS cells have immature osteoblastic properties and POS cells have mature osteoblastic properties. Retinoids decreased osteocalcin production in all the osteosarcoma cells. They induced an increase in production of type I collagen in HOS and POS cells, but a decrease in OOS cells. These results indicate that retinoids induce differentiation of canine osteosarcoma cells, resulting in an altered expression of their malignant phenotype. PMID- 10684760 TI - Presence of p53 mutations in feline neoplasms. AB - A region from exon 4 to 8 of the tumour suppressor gene p53 was analysed in 60 feline tumours (30 fibrosarcomas, seven malignant histiocytomas, three lymphosarcomas, five basal cell tumours, five squamous cell carcinomas, two adenocarcinomas of tubular skin glands, one undifferentiated carcinoma of the skin, seven mammary carcinomas). Missense mutations were detected in two fibrosarcomas, one malignant fibrous histiocytoma, the undifferentiated carcinoma of the skin and one mammary carcinoma. One nonsense mutation was detected in one fibrosarcoma and one deletion/frameshift-mutation was observed in one squamous cell carcinoma. PMID- 10684762 TI - Enteric colonisation following natural exposure to Campylobacter in pigs. AB - A survey was conducted to establish the prevalence of Campylobacter in pigs from an integrated commercial hog farm. This study was carried out in four different groups of pigs: 1) adult gilts (50); 2) pregnant sows (9); 3) piglets at day-of birth (73); 4) weaned piglets (20). Rectal and/or caecal samples were collected from each pig. Campylobacter was cultured and enumerated from such samples using Bolton enrichment broth and Campy-Cephex agar plates. Both biochemical and serological tests were used to determine Campylobacter species. Gilts had a 76 per cent incidence of Campylobacter with a mean of 76.3 per cent for C. jejuni, 21 per cent for C. coli and 2.6 per cent for C. lari. Pregnant sows had a 100 per cent incidence of Campylobacter with a mean of 87 per cent for C. jejuni and 13 per cent for C. coli. Newborn piglets had a 57. 8 per cent incidence of Campylobacter, rising to 100 per cent by the time of weaning. Thus it appears that pigs, from the day of birth, are highly susceptible to colonisation by Campylobacter. PMID- 10684761 TI - Effect of lead on erythrocytic antioxidant defence, lipid peroxide level and thiol groups in calves. AB - Fifteen crossbred male calves were exposed to lead for a period of 28 days orally at the dose rate of 7.5 mg of lead acetate as 0.75 per cent solution kg(-1)body weight to study its effect on erythrocytic antioxidant defense, lipid peroxide level and thiol groups. Five calves were given no treatment and served as unexposed controls. Blood samples were collected before exposure to lead and thereafter at weekly intervals (ie. on day 7, 14, 21 and 28). Erythrocyte haemolysate (10 per cent) was prepared and analysed for lipid peroxide level, activity of Superoxide dismutase (SOD) and catalase. Total, protein-bound and non protein-bound thiol groups were also measured. Exposure to lead significantly (P < 0.05) reduced the erythrocytic SOD activity by day 7 and it remained lower until day 21 followed by a marginal increase on day 28. Catalase activity declined after an initial compensatory rise on day 7. Erythrocytic lipid peroxide level was recorded to be significantly (P < 0.05) higher by day 21 and 28 of exposure. Total, protein-bound and non protein-bound -SH content of erythrocytes declined. It was concluded that oral exposure of lead reduced the erythrocytic thiol content and antioxidant defence indicating possible role of free radicals in pathogenesis of lead toxicity. PMID- 10684763 TI - Inhibitory effects of 22-oxa-calcitriol and all- trans retinoic acid on the growth of a canine osteosarcoma derived cell-line in vivo and its pulmonary metastasis in vivo. AB - Pulmonary metastasis is a major cause of death and a major obstacle to the successful treatment of canine osteosarcoma. However, the residual capacity of the neoplasia for differentiation and its susceptibility to undergo apoptosis may be used to suppress its growth and metastatic properties. The highly metastasizing POS (HMPOS) canine osteosarcoma cell line which preferentially metastasize to the lungs was used to test the possible efficacy of 22-oxa calcitriol (OCT) and all- trans retinoic acid (ATRA) to inhibit growth and pulmonary metastasis of the subcutaneously grown osteosarcoma in nude mice. Treatments in vitro, morphologically elongated and increased alkaline phosphatase activity and staining of cells. Tumour growth in vivo was inhibited significantly and the combination treatment of OCT and ATRA (OCT + ATRA) exerted a synergistic and stronger suppression at concentration of 1.0 microg kg(-1)body weight when given subcutaneously three times a week for 5 weeks. The subcutaneous tumours of the control mice consisted of osteoblast-like cells and isolated chondroblast like cells, but formed several areas of osteoid and increased amount of collagen tissue in all treated mice. Pinpoint macrometastatic nodules developed only in all control mice. Micrometastatic nodule developed only in two of six mice treated with ATRA. However, nodule size and number, and lung wet weight were all reduced significantly. Metastasis were not seen in the mice treated with OCT or OCT + ATRA. This study demonstrated that inhibition of growth and pulmonary metastasis was induced by subcutaneous treatment with these drugs and suggest that both its differentiating and apoptotic inducing activities may be responsible for the antitumour effects. These drugs may be useful in the clinic as an adjunct for the treatment of canine osteosarcoma. PMID- 10684764 TI - Tissue sources of serum alkaline phosphatase in 34 hyperthyroid cats: a qualitative and quantitative study. AB - The concentration of serum alkaline phosphatase (SALP) is commonly elevated in hyperthyroid cats. Agarose gel electrophoresis, in tris -barbital-sodium barbital buffer, with and without the separation enhancer neuraminidase, was used to investigate the sources of the constituent isoenzymes of SALP in serum samples from 34 hyperthyroid cats, comparing them to sera from five healthy cats and to tissue homogenates from liver, kidney, bone and duodenum. Contrary to previous reports, treatment of serum with neuraminidase made differentiation of the various isoenzymes more difficult to achieve. A single band corresponding to the liver isoenzyme (LALP) was found in 100 per cent of healthy cats. Eighty-eight per cent of the hyperthyroid cats showed two bands, corresponding to the liver and bone (BALP) isoenzymes while 12 per cent showed a LALP band alone. In hyperthyroid cats, there was a significant correlation between the serum L thyroxine concentrations and the SALP concentrations. These findings suggest pathological changes in both bone and liver in most cases of feline thyrotoxicosis. PMID- 10684765 TI - Cell mediated and humoral immune responses of white-tailed deer experimentally infected with Mycobacterium bovis. AB - The objective of this study was to improve the understanding of immune responses of whitetailed deer (Odocoileus virginianus) infected with Mycobacterium bovis. Ten mature, female, white-tailed deer were inoculated by intratonsilar instillation of 2 x 10(3)or 2 x 10(5)colony-forming units of M. bovis. Lymphocyte proliferation and humoral response to M. bovis PPD and the M. bovis protein, MPB70 were measured. Deer were tested for exposure to M. bovis by the comparative cervical skin test. Biopsy specimens of skin test sites were examined microscopically and immunohistochemically. The comparative cervical skin test correctly identified all M. bovis -inoculated deer as exposed to M. bovis. Lymphocyte proliferative responses to MPB70 were more consistent than responses to M. bovisPPD in M. bovis -inoculated deer. Antibody responses were more prominent in deer with disseminated disease than in deer with localised disease. The cellular components of delayed-type hypersensitivity reactions at skin test sites were similar to tuberculin reactions in other species. T lymphocytes of the gamma/delta phenotype were seen in increased numbers in M. bovisPPD injection sites. PMID- 10684766 TI - Tissue print hybridisation and reverse transcriptase PCR in the detection of infectious bursal disease viruses in bursal tissues. AB - The genome segments of infectious bursal disease viruses (IBDV) in the bursa of Fabricius from experimently infected chickens or field samples were detected by tissue print hybridization (TPH) with subsequent reverse transcriptase (RT)- polymerase chain reaction (PCR). Bursae were imprinted onto nylon membrane and then hybridized with a cloned digoxigenin (DIG)-labeled cDNA probe. Tissue prints on nylon membrane were readily distinguished from control prints by color development and differences in signal intensity. In order to verify the TPH test, RT - PCR was used to amplify a 643-base pair fragment on the VP 2 gene of IBDV in the bursa of Fabricius. With all isolates, a c DNA fragment of 643 bp long was generated as expected and further confirmed the specificity of TPH. Our results suggest that a large number of field samples or selected tissues can be rapidly examined by TPH technique when combined with a cloned DIG -labeled c DNA probe. PMID- 10684767 TI - A tribute to the medical council on alcoholism, its journal and their supporters as we enter the new millennium PMID- 10684768 TI - The Medical Council on Alcoholism (MCA) and its journal Alcohol and Alcoholism. PMID- 10684769 TI - ESBRA: passport for the development of biological research on alcoholism in Europe. PMID- 10684770 TI - One hundred years of alcoholism: the Twentieth Century. AB - The past 100 years witnessed the formation of a disease concept of alcoholism and a rapid increase in the knowledge of its aetiopathology and treatment options. In the first half of the century, public sanctions aimed at the abolition of alcoholism. In the United States, alcohol prohibition was revoked in the economic turmoil of the Great Depression. In Germany, proposed medical procedures to reduce the fertility of alcoholics had catastrophic consequences during the fascist dictatorship. A revived focus on alcoholics as patients with a right to medical treatment came out of self-organized groups, such as Alcoholics Anonymous. The current disease concept includes the psychosocial and neurobiological foundations and consequences of alcoholism. Neurobiological research points to the dispositional factor of monoaminergic dysfunction and indicates that neuroadaptation and sensitization may play a role in the maintenance of addictive behaviour. New treatment options include pharmacological approaches and indicate that behaviour and motivational therapy and the attendance of patient groups may equally reduce the relapse risk. The task of the future will be to apply scientific discoveries in the best interest of the patients and to support their efforts to be respected like subjects suffering from other diseases. PMID- 10684771 TI - Science, practice and patient needs: the work of the Plinius Maior Society. AB - The Plinius Maior Society is a European multinational, multidisciplinary group of clinicians and researchers in the alcoholism field, which strives for a comprehensive care concept in the management of alcoholism and alcohol-related problems. The Society, using evidence-based medicine, has developed a set of protocols, in the forms of guidelines, flow-charts, leaflets and booklets, for use as tools in research on and treatment of alcohol dependence, with a view to standardize clinical research procedures and to bridge the gap between the alcoholism researcher, practitioner and patient. These protocols or tools have been subjected to a review process during their preparation, and further comments on their validity will be integrated in their updates. Seven protocols have so far been developed, two of which, 'Guidelines on Evaluation of Treatment of Alcohol Dependence' and 'Detection and Management of Patients with Psychiatric and Alcohol Use Disorders', are aimed at the clinical researcher and specialists, whereas three others [in the form of decision trees (flow-charts)] are aimed at the general practitioner and other primary health care providers. These are entitled 'Alcohol Risk Assessment and Intervention in Primary Care', 'Withdrawal from Alcohol at Home' and 'Brief Intervention in Patients with Alcohol-Related Problems'. The remaining two tools are booklets aimed at the patient, one to support initiatives for detection of drinking problems and primary intervention, namely 'Do you have this Problem? Discuss it with your Doctor!', and the other to assist the patient in relapse prevention after the early stages of treatment, namely 'On the Way to Recovery'. The protocols for the general practitioners and patients have so far been produced in seven European languages, and, as with the Guidelines, feedback from target users will be collected and incorporated in future updates. The Society continually seeks to consider areas of clinical importance for its work and, as it enters the new millennium, it hopes to address and make a significant contribution to the most pressing problem in the management of alcohol dependence, namely relapse. PMID- 10684772 TI - Developmental aspects of intestinal intraepithelial and lamina propria lymphocytes in the rat following placental and lactational exposure to ethanol. AB - Fetal and lactational exposure to alcohol can induce impairments to the immune system and lead to decreased resistance to certain infectious agents. Morphometric procedures were used to quantify changes induced by maternal ethanol consumption in the gut-associated lymphoid tissue of rat pups. Rats were pair-fed with ethanol-containing or isocaloric control liquid diets formulated for pregnant and lactating animals from day 1 of pregnancy and throughout the lactation periods. Pups were weaned and placed on control liquid diet on post natal day 21. Intraepithelial and lamina propria lymphocytes and macrophages were evaluated on post-natal days 14, 18, and 25. Lower thymus weights were observed in the ethanol-exposed pups on post-natal days 14 and 18 and lower total thymocyte counts on post-natal day 14. On post-natal day 14, T cells, T cytotoxic cells, IgA plasma cells, and macrophages were decreased in the ileal epithelial and lamina propria areas in the ethanol-exposed, compared to the pair-fed, pups. No differences for any of the above cell counts were found in the jejunum on post natal day 14. On post-natal day 18, macrophages were still decreased in the ileum in the ethanol-treated pups, compared to pair-fed animals. No differences were found in T cells, T cytotoxic cells, and IgA lymphocytes between groups in either the jejunum or ileum on post-natal days 18 and 25. This study suggests that fetal and lactational exposure to ethanol has some effects on the development or influx of intraepithelial and lamina propria leukocytes and that the changes are most pronounced in early neonatal life. PMID- 10684773 TI - Role of Kupffer cells in the release of nitric oxide and change of portal pressure after ethanol perfusion in the rat liver. AB - The objective of this study was to elucidate the role of Kupffer cells during the increase of portal vein pressure caused by ethanol. We measured nitric oxide (NO) in the perfused rat liver using a commercial NO meter. Ethanol perfusion increased NO release and portal vein pressure. Gadolinium chloride pretreatment reduced the increase in portal vein pressure during the early phase of ethanol perfusion, but did not affect the release of NO after ethanol infusion. These findings suggest that Kupffer cells play an important role in liver microcirculation during the early stage of ethanol intake, but that the mechanism may not be regulated by NO. PMID- 10684774 TI - Cerebellar Purkinje neurons with altered terminal dendritic segments are present in all lobules of the cerebellar vermis of ageing, ethanol-treated F344 rats. AB - Previous studies from this Laboratory have shown that cerebellar Purkinje neurons (PN) in ageing, ethanol-fed Fischer 344 rats may have terminal dendritic segments that are longer than in control rats. They also showed that the longer terminal segments represented a toxic effect of ethanol on PN, because their increase in length resulted from an ethanol-induced deletion of other terminal dendritic segments and not from dendritic growth. The purpose of the present study was to determine whether this effect of ethanol was localized to specific lobules or was widely distributed within all lobules of the cerebellar vermis. Twelve-month-old male Fischer 344 rats were treated with a liquid ethanol diet for 48 weeks. Age- and weight-matched controls were pair-fed with an isocaloric control diet. Terminal dendritic segments in Golgi-Cox-stained PN in four groups of lobules in the cerebellar vermis of control and ethanol-fed rats were measured for treatment related changes in length. Results from these measurements showed that ethanol exposed PN with significantly longer terminal segments were present in all groups of lobules, i.e. they were widely distributed and not confined to specific lobules. Results from these measurements also confirmed in a large sample of neurons (40 neurons per rat) that the topologically distinct unpaired terminal segments were characteristically longer than the paired terminal segments in PN of control and ethanol-fed rats and that both types of terminal segments were longer in the ethanol-fed rats than in the controls. PMID- 10684775 TI - Effects of carnosine and related compounds on the stability and morphology of erythrocytes from alcoholics. AB - The effects of carnosine and related compounds on erythrocytes from alcoholics were studied. In their presence, erythrocytes showed an increased ability to resist haemolysis and showed a more normal morphology, with carnosine and N acetyl-carnosine being the most effective compounds. These beneficial properties of the dipeptides do not appear to be directly related to their antioxidant or buffering properties. PMID- 10684776 TI - A pilot investigation of the effect of tryptophan manipulation on the affective state of male chronic alcoholics. AB - A pilot study was conducted to investigate the hypothesis that dietary tryptophan manipulation would influence self-report affective status in alcoholic males. No significant effect of dietary manipulation was observed on the tryptophan/large neutral amino acids ratio or psychological indices of affect. The notion that dietary manipulation may be utilized in improving mood state in alcoholic males was not supported. PMID- 10684777 TI - Dissociable cognitive impairments in problem drinkers. AB - Patients in a treatment programme for severe alcohol dependence were tested on a battery of tests designed to examine organizational and visuo-spatial abilities. Analysis using a case-study approach indicated independent organizational and visuo-spatial impairments. An understanding of aetiological factors underlying these cognitive deficits and implications for treatment are discussed. PMID- 10684778 TI - Application of cross-impact analysis to the relationship between aldehyde dehydrogenase 2 allele and the flushing syndrome. AB - The experimental approach has been used to study successfully the relationship between the polymorphism of aldehyde dehydrogenase 2 and the flushing syndrome. In the present study, a probabilistic approach was used to analyse this relationship. Using cross-impact analysis and experimental data, the probability of the occurrence of flushing, the extent by which ALDH2*2/*2 enhances flushing more than ALDH2*1/*2, the extent by which ALDH2*1/*1 enhances non-flushing more than ALDH2*1/*2, etc. can be calculated. Two examples are given to show the potential use of cross-impact analysis. PMID- 10684779 TI - A comparison of substance use between female inmates and female substance misusers in treatment. AB - Recent literature documents extensive substance misuse histories among US female prison inmates. The primary purpose of the present study was to determine whether histories of personal and familial substance misuse distinguished female inmates from substance misusers in treatment. After accounting for drug-related offences, we hypothesized that the inmates would have more extensive histories of personal and familial substance misuse and that they would have initiated substance use at an earlier age. Contrary to our expectations, the two samples were similar on many measures of alcohol and drug use. Similarly, differences in family histories of substance misuse were not in the predicted direction. As hypothesized, however, the inmates did report earlier age at onset of drinking. Of particular clinical relevance was the finding that, despite similar alcohol consumption levels, inmates reported fewer alcohol-related adverse medical, legal, and psychosocial consequences than did the treatment sample. PMID- 10684780 TI - Detoxification from alcohol: a comparison of home detoxification and hospital based day patient care. AB - An uncontrolled study was carried out to examine two types of ambulatory care for patients undergoing detoxification from alcohol. The safety, efficacy, and acceptability of home detoxification was compared to detoxification within a day hospital setting. Seventy-nine per cent of home detoxification patients, many of whom had major alcohol-related problems and were severely dependent on alcohol, were successfully detoxified at 10 days. The day hospital group overlapped in severity with the home group and 78% completed detoxification. At 60 days, 45% of home detoxification patients and 31% of the day hospital group showed significant improvements in a range of alcohol-related difficulties. Improved outcome was associated with attendance for further treatment for both groups. Both home and day hospital detoxifications were viable alternatives to in-patient detoxification for selected groups of patients. PMID- 10684781 TI - Suicides of alcohol misusers and non-misusers in a nationwide population. AB - Alcohol dependence is a risk factor for suicide, and in the general population alcohol consumption and suicide rates are known to be associated. We investigated victims with and without alcohol misuse among unselected completed suicides to explore the role of alcohol misuse in the suicidal process and final act. In a total 1-year (1987-1988) population of suicides in the National Suicide Prevention Project in Finland, alcohol-misusing and -non-misusing victims were compared. On the basis of informant interviews, 35% (n = 349) of included victims were classified as alcohol misusers and 65% (n = 648) as non-misusers. The misusers were more often younger, male, divorced or separated and had more often worked, but were recently unemployed. They had experienced more often recent adverse life events possibly dependent on their own behaviour, were far more likely to be alcohol-intoxicated at the time of suicide, and tended to die from drug overdose. Several characteristics of these predominantly male alcohol misusers indicated better earlier lifetime psychosocial adjustment compared to the non-misusers, but more adverse life events close to suicide. Alcohol misuse is likely to have a deteriorating influence on the life course of those who eventually succumb to suicide, and its adverse consequences are common in misusers during the final months. PMID- 10684782 TI - Combination pharmacotherapy: a mixture of small doses of naltrexone, fluoxetine, and a thyrotropin-releasing hormone analogue reduces alcohol intake in three strains of alcohol-preferring rats. AB - It is common to treat some diseases with more than one medication simultaneously. Since more than one neurotransmitter system is involved in alcohol-seeking behaviour, then a therapeutic approach that targets more than one system should be more effective in reducing alcohol intake than one addressing a single system. To test this hypothesis, we compared the efficacy of low doses of individual drugs reported to reduce voluntary alcohol drinking to the efficacy of a mixture of these agents at the same low doses in reducing alcohol intake in three strains of alcohol-preferring rats (P, HAD, and Fawn-Hooded). After establishment of a stable baseline for alcohol intake in a continuous access paradigm, each rat received separate single i.p. injections of relatively low doses of either naltrexone (2.0 mg/kg), fluoxetine (1.0 mg/kg), the thyrotropin-releasing hormone analogue TA-0910 (0.2 mg/kg), a mixture of all three drugs, or the vehicle at 09:30. Each rat received all treatments, with an inter-injection washout period of at least 3 days. Alcohol and water intakes were measured at 6 and 24 h, and food intake was measured at 24 h, after the injection. Our results show that individual drugs did not significantly affect food, water, or alcohol intake. However, the mixture significantly reduced alcohol intake in all three strains, but had no effect on food intake. Similar results were obtained when the HAD rats received an oral dose of the individual drugs or the mixture. When P rats were given an i.p. injection of the mixture for 10 consecutive days, there was a continued suppressing effect. These findings show that a combination treatment designed to target simultaneously serotonergic, dopaminergic, and opioidergic systems can reduce alcohol intake, even though the doses of the individual drugs in the mixture are relatively low and ineffective when given singly. PMID- 10684783 TI - Acute effect of alcohol on androgens in premenopausal women. AB - The aim of the present study was to investigate the effect of alcohol on androgen levels among premenopausal women. Eighty-seven women in the mid-cycle phase of the menstrual cycle, 47 of whom used oral contraceptives (OC+), were included in the study. The range for reported alcohol consumption was 0-4 drinks/day. The total testosterone levels were significantly higher after alcohol intake (0.5 g/kg) than after placebo at 45 min and 90 min from the start of drinking among both OC- and OC+ subjects. This effect was also seen in the free testosterone fraction. The effect on testosterone was more prominent among OC+ subjects. Androstenedione levels were significantly lowered and the testosterone:androstenedione ratio significantly elevated by alcohol among both OC- and OC+ subjects. No effect of alcohol on dehydroepiandrosterone or dihydrotestosterone levels was observed. A positive correlation was observed between the change in testosterone levels and the change in androstenedione levels during placebo conditions. The correlation was significantly reduced during alcohol conditions among OC+ subjects, indicating an increased androstenedione to testosterone conversion. No significant dose (0.34, 0.68 and 1.02 g/kg) or time (45, 90 and 150 min) effects on total testosterone were observed in a substudy involving 10 OC+ subjects. The present results suggest that the testosterone effect is related to the zero-order mechanism of ethanol oxidation. The observed testosterone and androstenedione effects are suggested to be the result of an increased androstenedione to testosterone conversion in the liver caused by the alcohol-mediated elevation in the [NADH]:[NAD(+)] ratio. The present findings may be relevant in the development of hyperandrogenism and loss of female sexual characteristics associated with heavy alcohol consumption. PMID- 10684784 TI - Naltrexone exerts a favourable effect on plasma lipids in abstinent patients with alcohol dependence. AB - Epidemiological studies suggest that abstinence periods in some patients with alcohol dependence may increase their cardiovascular risk via proatherogenic changes in plasma lipid levels. Because of this, drugs administered in withdrawal therapy should not exacerbate these effects. The aim of this study was to estimate the influence of naltrexone, carbamazepine, and lithium carbonate on plasma lipid levels in 160 alcohol-dependent males during withdrawal therapy. Plasma concentrations of total cholesterol (TC), HDL cholesterol (HDL-C), LDL cholesterol (LDL-C), and triglycerides (TGL) were determined every 2 weeks for 20 weeks. Pharmacotherapy (naltrexone 50 mg, carbamazepine 600-800 mg, lithium carbonate 500-1000 mg once per day or placebo) was given within the framework of a double-blind study between the fourth and twentieth weeks of the study. The results of 116 patients who maintained abstinence during the whole 20-week observation period were analysed. In patients treated with naltrexone significant decreases in TC (239 +/- 58 vs 216 +/- 52 mg/dl; P < 0.01) and TGL (125 +/- 68 vs 86 +/- 33 mg/dl; P < 0.02) concentrations after 16 weeks of pharmacotherapy were observed. In patients treated with carbamazepine, significant increases in TC (224 +/- 39 vs 243 +/- 54 mg/dl, P < 0.04) and HDL (40 +/- 10 vs 44 +/- 8 mg/dl, P < 0.01) after 16 weeks of pharmacotherapy were observed. After 16 weeks of pharmacotherapy, patients treated with naltrexone had lower mean TC (P < 0.03) and LDL-C (P < 0.01) concentrations than patients treated with carbamazepine, lower mean LDL-C levels than patients treated with lithium carbonate (149 +/- 54 vs 164 +/- 57 mg/dl, P < 0.01), and lower TGL concentrations than patients of the remaining pharmacotherapy groups. We conclude that naltrexone, by its hypolipaemic effect, could be useful for withdrawal therapy in alcoholic patients, because it may decrease the cardiovascular risk in abstinent patients with alcohol dependence by lipid mechanisms. PMID- 10684785 TI - Cytochrome P-450 2E1 activity and oxidative stress in alcoholic patients. AB - As cytochrome P-450 2E1 (CYP2E1) induction was related to oxidative stress in experimental models, the aim of this study was to investigate the relationship between CYP2E1 activity and markers of oxidative stress in 40 alcoholic patients entering a rehabilitation programme. Plasma oxidized proteins, lipid peroxides (LPO) and antibodies against hydroxyethyl radical (HER) or malondialdehyde (MDA) adducts were assessed as markers of the production of free radicals, whereas vitamin E levels were evaluated as a marker of the antioxidant defence. CYP2E1 activity was determined by using the 6-hydroxychlorzoxazone:chlorzoxazone blood metabolic ratio, 2 h after drug intake. This ratio was increased by 4-fold in alcoholics, compared to non-alcoholic patients, and was correlated with daily intake of ethanol, carbohydrate-deficient transferrin, and blood alcohol level at the time of admission to hospital. Plasma levels of LPO and oxidized proteins were slightly increased (20%) in alcoholic patients when compared with the control group, whereas those of vitamin E were found to be slightly decreased (by 18%). Antibodies against HER or MDA adducts showed a very significant increase. However, when alcoholic patients were divided into two groups according to low or high CYP2E1 induction, no significant difference was observed in the variation of these parameters, except for anti-HER adducts antibodies. Therefore, our study confirms the main involvement of CYP2E1 in HER production. By contrast, CYP2E1 does not appear to be the main factor responsible for the oxidative stress occurring during human chronic alcoholism. Free radicals from other sources may therefore contribute significantly to the generation of this oxidative stress. PMID- 10684786 TI - Self-estimates of blood-alcohol concentration and ability to drive in a population of soldiers. PMID- 10684787 TI - Role of transforming growth factor-alpha and the epidermal growth factor receptor in embryonic rat testis development. AB - Embryonic testis development requires the morphogenesis of cords and growth of all cell populations to allow organ formation. It is anticipated that coordination of the growth and differentiation of various cell types involves locally produced growth factors. The current study was an investigation of the hypothesis that transforming growth factor-alpha (TGF-alpha) is involved in regulating embryonic testis growth. TGF-alpha has previously been shown to function in the postnatal testis. TGF-alpha and other members of the epidermal growth factor (EGF) family act through the epidermal growth factor receptor (EGFR) to stimulate cell proliferation and tissue morphogenesis. To understand the potential actions of TGF-alpha in the embryonic testis, general cell proliferation was investigated. Characterization of cell proliferation in the rat testis throughout embryonic and postnatal development indicated that each cell type has a distinct pattern of proliferation. Germ cell growth was transiently suppressed around birth. Interstitial cell growth was high embryonically and decreased to low levels around birth. A low level of Sertoli cell proliferation was observed at the onset of testis cord formation. Sertoli cell proliferation in early embryonic development was low; the levels were high later in embryonic development and remained high until the onset of puberty. Both TGF-alpha and the EGFR were shown to be expressed in the embryonic and postnatal rat and mouse testis. Perturbation of TGF-alpha function using neutralizing antibodies to TGF alpha on testis organ cultures dramatically inhibited the growth of both embryonic and neonatal testis. TGF-alpha antibodies had no effect on cord formation. The TGF-alpha antibody was found to be specific for TGF-alpha in Western blots when compared to EGF and heregulin. Testis growth was also inhibited by perturbation of EGFR signaling using an EGFR kinase inhibitor. Therefore, TGF-alpha appears to influence embryonic testis growth but not morphogenesis (i.e., cord formation). Treatment of embryonic testis organ cultures with exogenous TGF-alpha also perturbed development, leading to an increased proliferation of unorganized cells. Testis from EGFR and TGF-alpha knockout mice were analyzed for testis morphology. TGF-alpha knockout mice had no alterations in testis phenotype, while EGFR knockout mice had a transient decrease in the relative amount of interstitial cells before birth. Observations suggest that there may be alternate or compensatory factors that allow testis growth to occur in the apparent absence of TGF-alpha actions in the mutant mice. In summary, the results obtained suggest that TGF-alpha is an important factor in the regulation of embryonic testis growth, but other factors will also be involved in the process. PMID- 10684788 TI - Expression and action of hepatocyte growth factor in human and bovine normal ovarian surface epithelium and ovarian cancer. AB - More than 95% of ovarian cancers originate from the epithelial cells on the surface of the ovary, which are termed ovarian surface epithelium (OSE). These OSE cells are modified peritoneal mesothelial cells separated from underlying ovarian surface stromal tissue by a basal lamina of dense collagenous connective tissue. Mesenchymal-epithelial cell interactions between stromal cells and OSE cells are postulated to be important for normal OSE biology and for the onset of ovarian cancer. Hepatocyte growth factor (HGF) is a mesenchymal-derived growth factor that mediates mesenchymal-epithelial cell interactions in a number of different tissues. The current study was an investigation of the expression and actions of HGF in normal OSE and ovarian cancer. Human epithelial cells from borderline and stage III ovarian cancer cases were found to express HGF protein in the epithelial cell component by immunocytochemistry analysis. The stromal cell component of human ovarian tumors contained little or no HGF immunostaining. Normal bovine ovaries have a similar physiology and endocrinology to human ovaries and are used as a model system to investigate normal OSE functions. HGF protein was detected in the OSE from both normal human and bovine ovaries. Adjacent ovarian stromal tissue contained light but positive HGF immunostaining. RNA was collected from normal bovine ovarian stromal cells to examine HGF gene expression. HGF transcripts were detected in cultured OSE and stromal cells by Northern blot analysis. Using a quantitative reverse transcription-polymerase chain reaction procedure, HGF gene expression was found to be high in freshly isolated OSE but low in freshly isolated stroma. Levels of HGF gene expression after culture of stroma increased. Observations indicate that normal OSE express high levels of HGF in vivo and in vitro. Expression of HGF by normal epithelial cells versus stromal cells was unexpected and suggests that HGF may be important in an autocrine regulation of OSE. HGF actions on normal OSE cells and ovarian cancer cells were investigated. HGF was found to stimulate the growth of normal OSE cells in a manner similar to such growth stimulated by epidermal growth factor. Two ovarian cancer cell lines, SKOV3 and OCC1, were also stimulated to grow in response to HGF. This observation suggests that HGF may be involved in sustaining growth of ovarian tumors. These results are the first to demonstrate the production and action of HGF in normal OSE cells and ovarian cancer cells. This appears to be an example of HGF production by an epithelial cell, such that a mesenchymal-epithelial mixed phenotype is present. The autocrine stimulation of OSE growth by the local production and action of HGF provides insight into how the OSE may develop abnormal growth characteristics involved in the onset and progression of ovarian cancer. PMID- 10684789 TI - Identification and localization of plasminogen activator inhibitor-1 within the porcine oviduct. AB - The porcine oviduct synthesizes de novo and secretes a number of proteins into culture medium, many of which are unidentified. The objectives of the present study were to 1) semipurify and identify a M(r) 45 000 secreted protein of the oviduct, 2) examine its synthesis within the three functional segments (infundibulum, ampulla, and isthmus), and 3) evaluate its distribution throughout the oviduct. Oviductal tissue was collected during early pregnancy, divided into functional segments, and subsequently cultured. Medium was collected, and the M(r) 45 000 protein was concentrated by gel-filtration chromatography. The semipurified protein was transferred onto a polyvinylidene fluoride membrane and subjected to N-terminal amino acid analysis. The 26-amino acid sequence was 96% identical to that of pig plasminogen activator inhibitor (PAI)-1. Analysis by 1 dimensional SDS-PAGE and fluorography of rabbit anti-human PAI-1 immunoprecipitated product confirmed PAI-1. Subsequent 2-dimensional SDS-PAGE and fluorographic analyses of media revealed greater PAI-1 synthesis by the isthmus than by the ampulla or infundibulum. PAI-1 was immunolocalized throughout the oviduct and was heavily concentrated in the apical region of epithelial cells. Immunogold electron microscopy localized PAI-1 within putative secretory granules in the epithelial apical region and also associated with cilia in the isthmus. Isthmic PAI expression suggests a crucial role in protecting the preimplantation embryo from proteolytic degradation as well as in regulation of extracellular matrix turnover and remodeling. PMID- 10684790 TI - Sperm antigen 6 is the murine homologue of the Chlamydomonas reinhardtii central apparatus protein encoded by the PF16 locus. AB - A cDNA encoding sperm antigen 6 (Spag6), the murine homologue of the Chlamydomonas reinhardtii PF16 protein-a component of the flagella central apparatus-was isolated from a mouse testis cDNA library. The cDNA sequence predicted a 55.3-kDa polypeptide containing 8 contiguous armadillo repeats with 65% amino acid sequence identity and 81% similarity to the Chlamydomonas PF1 protein. An antipeptide antibody generated against a C-terminal sequence recognized a 55-kDa protein in sperm extracts and localized Spag6 to the principal piece of permeabilized mouse sperm tails. When expressed in COS-1 cells, Spag6 colocalized with microtubules. The Spag6 gene was found to be highly expressed in testis and was mapped using the T31 radiation hybrid panel to mouse chromosome 16. Mutations in the Chlamydomonas PF16 gene cause flagellar paralysis. The presence of a highly conserved mammalian PF16 homologue (Spag6) raises the possibility that Spag6 plays an important role in sperm flagellar function. PMID- 10684791 TI - Selective requirement for Cdc25C protein synthesis during meiotic progression in porcine oocytes. AB - Fundamental differences between meiosis and mitosis suggest that the shared central cell cycle machinery may be regulated differently during the two division cycles. This paper focuses on unique features of Cdc25C protein function during meiotic progression. We report on the existence of oocyte-specific CDC25C transcripts that differ from their somatic counterparts in the 3' untranslated region. While CDC25C mRNA levels remain constant in fully-grown oocytes, corresponding protein levels increase progressively during maturation to a maximum at metaphase II. Elevation of Cdc25C protein levels in G2-oocytes by mRNA injection failed to increase MPF-kinase levels or to induce premature entry into M-phase. Likewise, antisense-induced arrest of translation (translational arrest) had no effect on chromosome condensation, nucleolar disassembly, or nuclear membrane contraction. By contrast, translational arrest inhibited subsequent events including membrane disassembly and spindle formation. Neither up- nor down regulation of Cdc25C synthesis after metaphase I plate formation influenced progression to metaphase II. However, translational arrest during metaphase resulted in incomplete chromosome decondensation and abnormal pronuclear membrane assembly after activation. We conclude that Cdc25 protein, translated from unique transcripts, is preferentially located in the oocyte nucleus and is essential for progress through late diakinesis. Subsequently, new synthesis of Cdc25C protein is required for the orderly transition from meiotic to mitotic cell division. PMID- 10684792 TI - Pregnancy and exogenous steroid treatments modulate the expression of relaxant EP(2) and contractile FP receptors in the rat uterus. AB - Prostaglandins (PGs) interact with specific receptors on plasma membranes to regulate myometrial activity in many species. The present study examined whether the expression of relaxant prostaglandin E receptor subtype two (EP(2)) and contractile prostaglandin F receptor (FP) mRNA in the rat uterus is changed during various states of pregnancy and regulated by steroid hormones. Expression of mRNA for EP(2) and FP receptors in the full-thickness uteri was analyzed by reverse transcription-polymerase chain reaction using specific primers. Abundance of receptor mRNA was expressed relative to beta-actin mRNA. Results showed that 1) mRNA for EP(2) receptors in the rat uterus was substantially increased during pregnancy (320%) compared with the nonpregnant state (100%, P < 0.01), and declined during labor at term (36% vs. 100% in control, P < 0.01); 2) mRNA expression for FP receptors in rat uterus was increased during pregnancy (333% vs. 100% in nonpregnant rats, P < 0. 01) and reached maximal levels during labor (515% vs. 100% in control, P < 0.01); 3) upon RU-486 treatment on Day 19 of pregnancy, uterine EP(2) receptor mRNA levels were decreased (18% vs. 100% in control, P < 0.01), and FP mRNA levels were increased (357% vs. 100% in control, P < 0.01); 4) with ICI 164384 (an antiestrogen) treatment on Day 19 of gestation, uterine FP receptor mRNA levels were decreased without effects on EP(2) receptors; 5) in ovariectomized (ovx) rats, progesterone increased EP(2) (163% vs. 100% in control, P < 0.01) and had no effects on FP receptor mRNA expression in the rat uterus; 6) estradiol increased FP receptor mRNA levels (358% vs. 100% in control, P < 0.01) and had no effects on EP(2) mRNA in the ovx rat uterus. Therefore, we conclude that steroid hormones modulate the mRNA for relaxant EP(2) and contractile FP receptors for PGs in the uterus and thus regulate uterine activity during pregnancy and labor. PMID- 10684793 TI - Platelet-activating factor induces an imbalance between matrix metalloproteinase 1 and tissue inhibitor of metalloproteinases-1 expression in human uterine cervical fibroblasts. AB - Platelet-activating factor (PAF) is involved in such reproductive processes as parturition. We investigated the effect of PAF on the expression of matrix metalloproteinase-1 (MMP-1) and that of tissue inhibitor of metalloproteinases-1 (TIMP-1) in human uterine cervical fibroblasts. Uterine cervical tissue was obtained from patients who underwent cesarean section at term. Collagenase dispersed fibroblasts were cultured and used in the experiments. PAF receptor was identified in the uterine cervical fibroblasts by use of reverse transcription polymerase chain reaction and Southern blot analysis. Northern blot analysis showed that PAF increased the expression of MMP-1 mRNA in a time-dependent manner, whereas expression of TIMP-1 mRNA was not affected by PAF. Concentration of MMP-1 protein in the PAF-treated culture media significantly exceeded that in control cultures. The PAF-induced production of MMP-1 protein was abolished by treatment with WEB 2170, a specific PAF receptor antagonist. Results suggest that PAF may accelerate collagenolysis in the human uterine cervix by inducing an imbalance in the activity between MMP-1 and TIMP-1, thus contributing to the cervical ripening during parturition. PMID- 10684794 TI - Immunocytochemical localization of lipocalin-type prostaglandin D synthase in the bull testis and epididymis and on ejaculated sperm. AB - Previously, we identified a 26-kDa fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase. The objective of the present study was to immunohistochemically localize this enzyme to the various cell types within the bull testis and seven subsegments of the epididymis, and on ejaculated sperm in order to gain further insight into its potential function in male reproduction. In the testis, immunoperoxidase staining was localized within the elongating spermatids and Sertoli cells of the seminiferous tubules, varying with the stage of the spermatogenic cycle. The highest level of staining occurred during stages III-VII. The cuboidal epithelial cells of the rete testis and efferent ducts were also immunoreactive. Expression of lipocalin-type prostaglandin D synthase was not uniform in the seven epididymal subsegments, suggesting a possible role in sperm maturation. In all epididymal regions, expression was limited to the epithelial principal cells; no immunoreactivity was apparent in other cell types. Lipocalin-type prostaglandin D synthase was strikingly localized in the caput epididymidis, while moderate to weak staining was observed in the remainder of the epididymis. Droplets of reaction product observed within the lumen increased progressively from the caput to cauda. Using fluorescence microscopy, we also localized lipocalin-type prostaglandin D synthase to the apical ridge of the acrosome on ejaculated sperm. PMID- 10684795 TI - Sperm aster formation and pronuclear decondensation during rabbit fertilization and development of a functional assay for human sperm. AB - Microtubule organization and chromatin configurations in rabbit eggs after in vivo rabbit fertilization and after intracytoplasmic injection with human sperm were characterized. In unfertilized eggs, an anastral barrel-shaped meiotic spindle, oriented radially to the cortex, was observed. After rabbit sperm incorporation, microtubules were organized into a radial aster from the sperm head, and cytoplasmic microtubules were organized around the male and female pronuclei. The microtubules extending from the decondensed sperm head participated in pronuclear migration, and organization around the female pronucleus may also be important for pronuclear centration. Support for these observations was found in parthenogenetically activated eggs, in which microtubule arrays were organized around the single female pronucleus that formed after artificial activation. These observations support a biparental centrosomal contribution during rabbit fertilization as opposed to a strictly paternal inheritance pattern suggested from previous studies. In rabbit eggs that received injected human donor sperm, an astral array of microtubules radiated from the sperm neck and enlarged as the sperm head underwent pronuclear decondensation. gamma-Tubulin was observed in the center of the sperm aster. We conclude that the rabbit egg exhibits a blended centrosomal contribution necessary for completion of fertilization and that the rabbit egg may be a novel animal model for assessing centrosomal function in human sperm and spermatogenic cells following intracytoplasmic injection. PMID- 10684796 TI - Birth of piglets after transfer of embryos cryopreserved by cytoskeletal stabilization and vitrification. AB - Pig embryos suffer severe sensitivity to hypothermic conditions, which limits their ability to withstand conventional cryopreservation. Research has focused on high lipid content of pig embryos and its role in hypothermic sensitivity, while little research has been conducted on structural damage. Documenting cytoskeletal disruption provides information on embryonic sensitivity and cellular response to cryopreservation. The objectives of this study were to document microfilament (MF) alterations during swine embryo vitrification, to utilize an MF inhibitor during cryopreservation to stabilize MF, and to determine the developmental competence of cytoskeletal-stabilized and vitrified pig embryos. Vitrified morulae/early blastocysts displayed MF disruptions and lacked developmental competence after cryopreservation; hatched blastocysts displayed variable MF disruption and developmental competence. Cytochalasin-b did not improve morula/early blastocyst viability after vitrification; however, it significantly (P < 0.05) improved survival and development of expanded and hatched blastocysts. After embryo transfer, we achieved pregnancy rates of almost 60%, and litter sizes improved from 5 to 7.25 piglets per litter. This study shows that the pig embryo cytoskeleton can be affected by vitrification and that MF depolymerization prior to vitrification improves blastocyst developmental competence after cryopreservation. After transfer, vitrified embryos can produce live, healthy piglets that grow normally and when mature are of excellent fecundity. PMID- 10684797 TI - A metabolite of methoxychlor, 2,2-bis(p-hydroxyphenyl)-1,1, 1-trichloroethane, reduces testosterone biosynthesis in rat leydig cells through suppression of steady-state messenger ribonucleic acid levels of the cholesterol side-chain cleavage enzyme. AB - Postnatal development of Leydig cells involves transformation through three stages: progenitor, immature, and adult Leydig cells. The process of differentiation is accompanied by a progressive increase in the capacity of Leydig cells to produce testosterone (T). T promotes the male phenotype in the prepubertal period and maintains sexual function in adulthood; therefore, disruption of T biosynthesis in Leydig cells can adversely affect male fertility. The present study was designed to evaluate the ability of a xenoestrogen, methoxychlor (the methoxylated isomer of DDT [1,1, 1-trichloro-2,2-bis(p chlorophenyl)ethane]), to alter Leydig cell steroidogenic function. Purified progenitor, immature, and adult Leydig cells were obtained from, respectively, 21 , 35-, and 90-day-old Sprague-Dawley rats treated with graded concentrations of the biologically active metabolite of methoxychlor, 2, 2-bis(p-hydroxyphenyl) 1,1,1-trichloroethane (HPTE), and assessed for T production. HPTE caused a dose dependent inhibition of basal and LH-stimulated T production by Leydig cells. Compared to the control value, reduced T production by progenitor and immature Leydig cells was apparent after 10 h of HPTE treatment in culture; the equivalent time for adult Leydig cells was 18 h. The reversibility of HPTE-induced inhibition was evaluated by incubating Leydig cells for 3, 6, 10, 14, or 18 h and measuring T production after allowing time for recovery. After treatment with HPTE for 3 h, T production by immature and adult Leydig cells for the 18-h posttreatment period was similar to the control value, but that of progenitor Leydig cells was significantly lower. The onset of HPTE action and the reversibility of its effect showed that Leydig cells are more sensitive to this compound during pubertal differentiation than in adulthood. T production was comparable when control and HPTE-treated immature Leydig cells were incubated with pregnenolone, progesterone, and androstenedione, but HPTE-treated Leydig cells produced significantly reduced amounts of T when incubations were conducted with 22R-hydroxycholesterol (P < 0.01). This finding suggested that HPTE-induced inhibition of T production is related to a decrease in the activity of cytochrome P450 cholesterol side-chain cleavage enzyme (P450(scc)) and cholesterol utilization. The reduced steady-state mRNA level for P450(scc) in HPTE-treated Leydig cells was demonstrated by reverse transcription-polymerase chain reaction and densitometry. In conclusion, this study showed that HPTE causes a direct inhibition of T biosynthesis by Leydig cells at all stages of development. This effect suggests that reduced T production could be a contributory factor in male infertility associated with methoxychlor and, possibly, other DDT-related compounds. PMID- 10684798 TI - Assay and importance of adhesive interaction between hamster (Mesocricetus auratus) oocyte-cumulus complexes and the oviductal epithelium. AB - Adhesion between the oocyte-cumulus complex and infundibulum plays an important, but poorly understood, role in oocyte pick-up. The purposes of this study were to determine which components of the oocyte-cumulus complex and oviductal epithelium function in adhesion, to measure adhesion under physiological conditions, and to examine the effect of modulation of adhesion on oocyte-cumulus complex pick-up rate. Oocyte-cumulus complexes containing an expanded matrix were readily transported into the oviduct, while unexpanded complexes lacking an extracellular matrix were not picked up, indicating that the matrix is necessary for pick-up. Transmission electron microscopy revealed that during pick-up, adhesion occurred specifically between the ciliary crowns of the oviduct and the granules and filaments of the cumulus matrix. An assay was developed using vacuum from a low flow peristaltic precision pump, modified for bi-directional flow, to measure the strength of adhesion between the oocyte-cumulus complex and the oviductal epithelium, and adhesion was measured during physiological conditions. The lectin wheat germ agglutinin and the polycation poly-L-lysine were then used to modulate adhesion, and the effects of increasing or decreasing adhesion on oocyte pick-up rate and ciliary beat frequency were examined. The data show that 1) the matrix of the oocyte-cumulus complex and the ciliary crowns of the oviduct function in adhesion during pick-up and that adhesion is necessary for pick-up, 2) adhesion can be assayed quantitatively and is very uniform among control infundibula, and 3) decreasing or increasing adhesion decreases oocyte pick-up rate and in some cases prevents pick-up without affecting ciliary beat frequency. PMID- 10684799 TI - Caspase-3 and -6 expression and enzyme activity in hen granulosa cells. AB - We have cloned and sequenced cDNAs corresponding to the complete coding regions of the chicken homologues to mammalian caspase-3 and caspase-6. Both caspases are included among members of the cysteine protease (caspase) family that are most closely identified with mediating apoptosis. The deduced amino acid sequences for chicken caspase-3 and -6 show 65% and 68% identity with the respective human sequences, with complete conservation found within the QACRG active peptide region. Both caspase-3 and -6 are widely expressed within various tissues from the hen. Within the ovary, levels of caspase-3 and caspase-6 mRNA and protein do not change significantly in theca tissue during follicle development. On the other hand, procaspase-3 and -6 protein levels are elevated by 2- to 5-fold in preovulatory, compared to prehierarchal (6- to 8-mm diameter), follicle granulosa cells. Nevertheless, the function of this family of cell death-inducing proteins requires activation of the proenzyme caspase, which occurs after cleavage at predictable sites within the N-terminal domain. Accordingly, it was determined that okadaic acid, a pharmacologic inducer of apoptotic cell death in cultured apoptosis-resistant, preovulatory follicle granulosa cells, induced both caspase 3- and caspase-6-like activity within 8-16 h of treatment. By comparison, spontaneous apoptotic cell death that occurs in apoptosis-sensitive, prehierarchal follicle granulosa cells after short-term suspension culture is accompanied by a more rapid increase (within 2 h) in both caspase-3- and -6-like activity. Treatment with 8-bromo-cAMP, which has previously been shown to attenuate, or at least slow, the onset of apoptosis in prehierarchal follicle granulosa cells, mitigates this suspension culture-induced increase in caspase activity. While the present results provide further support for the relationship between caspase activation and apoptotic cell death in hen granulosa cells, the molecular ordering of enzymatic events and the caspase-specific substrates remain to be elucidated. PMID- 10684800 TI - Gonadotropin induction of ovulation and corpus luteum formation in young estrogen receptor-alpha knockout mice. AB - Estrogen receptor-alpha (ERalpha) knockout (ERalphaKO) female mice are infertile. Initially, they exhibit normal follicular development, but by 4-5 wk of age, they begin to develop hemorrhagic ovarian cysts. Follicles in adult ERalphaKO female mice progress to the graafian stage, but there are no corpora lutea (CL). To test whether ERalpha is required for ovarian folliculogenesis, ovulation, and CL formation, eCG and hCG were used to ovulate 3- to 5-wk-old ERalphaKO and wild type (WT) sibling mice. Gonadotropin administration resulted in ovulation in both ERalphaKO and WT mice. Gonadotropin-treated ERalphaKO females that ovulated produced 7.09 +/- 0.77 oocytes per mouse, whereas gonadotropin-treated WT female mice had 16.17 +/- 0.84 oocytes. Surprisingly, ruptured ERalphaKO ovarian follicles developed into CL that had normal morphology. Gonadotropin-treated ERalphaKO mice had 3-fold higher concentrations of serum progesterone than did control ERalphaKO mice that had been administered saline rather than gonadotropins. Thus, the CL in gonadotropin-treated ERalphaKO mice appeared to be steroidogenically functional. On the basis of these findings, ovarian folliculogenesis, ovulation, and CL formation can occur in the absence of ERalpha, although to a lesser extent than in WT mice. PMID- 10684801 TI - Expression of human proacrosin in Escherichia coli and binding to zona pellucida. AB - Proacrosin is a multifunctional protein present in the sperm acrosome. This study characterizes the expression of human proacrosin in bacteria and assesses zona pellucida binding activity. The cDNA encoding human proacrosin was subcloned in pGEX-3X and pET-22b vectors. In the pGEX system, expression of the full-length fusion protein was not detected. In the pET system, an expression product with an apparent molecular size similar to that expected for the proenzyme (Rec-40, 42-44 kDa) was recognized by a monoclonal antibody to human acrosin, AcrC5F10. A 32-34 kDa protein (Rec-30), not recognized by AcrC5F10 on Western blots, was the major expression product. Proteins of 21 (Rec-20) and 18 (Rec-10) kDa were recovered as insoluble expression products as were Rec-40 and Rec-30, and truncated products from the C terminus were detected in the soluble fraction. Rec-40 and Rec-30 coexisted at any culture time tested. Immune serum raised against Rec-30 (AntiRec 30) stained the acrosomal region of permeabilized human spermatozoa and recognized the recombinant proteins and proacrosin from human sperm extracts. Amino acid sequence analysis indicated that Rec-30, Rec-20, and Rec-10 are N terminal fragments of proacrosin. The recombinant proteins Rec-40, -30, -20, and 10 were found to interact with homologous (125)I-zona pellucida glycoproteins. PMID- 10684802 TI - Epigenetic modifications necessary for normal development are established during oocyte growth in mice. AB - The ability of maternal chromatin to support full-term development is attained during oocyte growth. The aim of this study was to identify when during the growth phase the maternal chromatin developed the capacity to support term development. Mature metaphase II-arrested oocytes that contained chromatin from oocytes at different stages of oocyte growth were constructed by micromanipulation. The oocytes were fertilized in vitro, developed to the blastocyst stage in vitro, and transferred to recipients to assay developmental potential. The results demonstrate, firstly, that the origin of the maternal chromatin has no effect on the rate of oocyte maturation, fertilization, or development to the blastocyst in vitro. Secondly we demonstrate that maternal chromatin is first competent to support development to term during the latter half of oocyte growth when oocytes are 60-69 microm in diameter in juvenile mice or 50-59 microm in diameter in adult mice. These data show that epigenetic modifications necessary for postimplantation development occur during a specific phase of oocyte growth. PMID- 10684803 TI - Interferon-tau and progesterone regulate ubiquitin cross-reactive protein expression in the ovine uterus. AB - Ubiquitin cross-reactive protein (UCRP) is a functional ubiquitin homolog synthesized by the ruminant endometrium in response to conceptus-derived interferon-tau (IFNtau). Progesterone is required for IFNtau to exert antiluteolytic actions on the endometrium. Therefore, this study was designed to determine whether progesterone is requisite for IFNtau induction of UCRP expression within the ovine uterus. Cyclic ewes were ovariectomized and fitted with intrauterine (i.u.) catheters on Day 5 and treated daily with steroids (i.m.) and protein (i.u.) as follows: 1) progesterone (P, Days 5-24) and control serum proteins (CX, Days 11-24); 2) P and ZK 137.316 (ZK; progesterone receptor antagonist, Days 11-24) and CX proteins; 3) P and recombinant ovine IFNtau (roIFNtau, Days 11-24); or 4) P and ZK and roIFNtau. All ewes were hysterectomized on Day 25. In P-treated ewes, roIFNtau increased endometrial UCRP mRNA and protein levels. However, administration of ZK to ewes ablated roIFNtau induction of UCRP. Recombinant ovine IFNtau induced expression of UCRP mRNA in progestinized endometrial luminal (LE) and glandular (GE) epithelium as well as in both stratum compactum and spongiosum layers of the stroma (ST). Progesterone receptor protein was located in endometrial ST, but not in LE and GE from these ewes. Results support the hypothesis that progesterone is required for IFNtau induction of type I IFN-responsive genes, such as UCRP, in the ruminant uterus. PMID- 10684804 TI - Assessing chromosomal abnormalities in two-cell bovine in vitro-fertilized embryos by using fluorescent in situ hybridization with three different cloned probes. AB - The aim of this study was to assess the efficiency of fluorescent in situ hybridization (FISH) for detecting chromosomal abnormalities in in vitro fertilized (IVF) bovine embryos as early as the 2-cell stage. Three different cloned probes were used, two derived from a unique sequence specific to the subtelomeric (D1S48) or subcentromeric regions (19C10) of chromosome 1 and the third (H1A clone) derived from a repetitive sequence that hybridizes to the subcentromeric regions of three other chromosomes (14, 20, 25). Our results show that the incidence of chromosomal abnormalities in 2-cell bovine IVF embryos varied from 28% to 44% according to the probes used for the analysis. Whereas the efficiency of FISH was high with somatic nuclei, it appeared to be highly variable with the 2-cell embryos. FISH efficiency depended firstly on the probe sequence (repetitive or unique sequence), secondly on the chromosomal target region (centromeric or telomeric regions), and thirdly on the embryo cell cycle phase. With a unique sequence probe (19C10) specific to the subcentromeric regions, FISH efficiency was better on nuclei in the S-phase cycle than on those in the G-phase. In S-phase 2-cell embryos, the overall incidence of chromosomal abnormalities was more accurately assessed. It reached 13% and was represented by 1n/2n mixoploidies. PMID- 10684805 TI - Follistatin suppresses steroid-enhanced follicle-stimulating hormone release in vitro in rats. AB - Previous in vitro and in vivo studies from our laboratory showed that progesterone (P(4)), corticosterone (B), and testosterone (T) increase intracellular content and release of FSH in the anterior pituitary. Activin (Act) and inhibin (Inh) are structurally related proteins with antagonistic actions, as Act stimulates and Inh inhibits FSH secretion from the anterior pituitary. Together with follistatin (FS), a protein that bioneutralizes Act, they form an autocrine-paracrine loop in the anterior pituitary that tightly regulates FSH secretion. The objective of the present study was to test the hypothesis that P(4), B, and T modulate this autocrine-paracrine loop to favor increased FSH secretion. If Act were to mediate steroid-induced FSH release, FS would be expected to block these effects. To test this interaction, cell cultures were prepared from anterior pituitaries of male and female rats, and treated with Act, B, P(4), or T in the absence or presence of FS. Act, B, P(4), and T increased FSH release; FS suppressed both basal and Act- and steroid-stimulated FSH release to approximately 50% below basal levels. Cell cultures from anterior pituitary of female rats were used to compare the interaction of incremental concentrations of FS on dose-related Act- and P(4)-stimulated FSH release. With increasing concentrations of Act, the FS-induced suppression of FSH release was attenuated and eventually abolished; in contrast, maximally stimulatory concentrations of P(4) did not fully overcome the FS-induced suppression of FSH release. The effects of P(4), B, and Act in the presence and absence of estradiol on steady state mRNA levels of FSHbeta, Actbeta(B), and FS were determined in primary pituitary cell cultures from metestrous female rats by reverse transcription polymerase chain reaction. Whereas Act, P(4), B increased FSHbeta mRNA levels, only Act raised the level of FS mRNA, and neither steroid increased Actbeta(B) mRNA. The results support the hypothesis that endogenous Act is a common mediator of the action of P(4), B, and T in the rat primary anterior pituitary cell culture. We conclude that the stimulation of FSH release and intracellular content in the gonadotroph by P(4), B, and T is mediated, in part, by Act and involves modulation of a tightly regulated Act/FS autocrine-paracrine loop. PMID- 10684806 TI - Vacuolar system-associated protein-60: a protein characterized from bovine granulosa and luteal cells that is associated with intracellular vesicles and related to human 80K-H and murine beta-glucosidase II. AB - It has been suggested that proteins of molecular size 56-58 kDa play an important role in bovine ovarian follicular development and oocyte maturation. A polyclonal antibody was raised against a 56- to 58-kDa protein band purified from bovine granulosa cells and was used to screen granulosa or luteal cell cDNA expression libraries. This work resulted in the identification of a cDNA encoding for a protein of 60.1 kDa with a signal peptide of 13 residues. The bovine 60.1-kDa protein shared an overall 86.7% and 81.8% identity with, respectively, the human 80K-H protein and the mouse putative beta subunit of glucosidase II (beta-GII), and was named vacuolar system-associated protein-60 (VASAP-60). Marked differences in sequence identity were noted in a putative molecular adapter domain containing a tandem D and E amino acid stretch flanked by proline-rich sequences presenting the minimal PXXP SH3 motif. VASAP-60 was shown to be unglycosylated using endoglycosidase H treatment and was found mainly in a cellular membrane fraction of bovine corpus luteum. VASAP-60 was localized in a rat hepatic Golgi/endosome fraction and in wheat germ agglutinin (WGA) affinity chromatographic eluates, thereby suggesting the presence of interactions with membrane glycoproteins. A polyclonal antibody was raised against the putative adapter domain of the recombinant VASAP-60; this was shown to recognize a major 88-kDa and two minor 58-kDa and 50-kDa proteins, suggesting that the major 88-kDa protein band represents the complete VASAP-60 protein whereas the 58-kDa and the 50-kDa bands represent its proteolytic fragments. Northern blot analysis demonstrated the presence of a single 2.3-kilobase transcript in all the bovine tissues analyzed with variation in the steady state level between tissues. Immunohistochemical observations showed that VASAP-60 was widely distributed in bovine tissues and was localized in pericytoplasmic and perinuclear membranes. In epithelial cells, the staining presented a basolateral or apical polarity associated with intracellular vacuoles. In conclusion, we have characterized a novel acidic membrane protein, associated with organelles of the vacuolar system, that is widely and histospecifically expressed in bovine tissues. VASAP-60 represents either the bovine ortholog or a new family member of the previously characterized human 80K-H and murine beta-GII proteins. Our results suggest that VASAP-60 presents characteristics of a molecular adaptor protein with functions in membrane-trafficking events. PMID- 10684807 TI - A putative stimulatory role of progesterone acting via progesterone receptors in the steroidogenic cells of the human corpus luteum. AB - To further explore the proposed auto-regulatory role of progesterone action in the human corpus luteum (CL), the expression and functional roles of progesterone receptor (PR) isoforms A and B during the luteal phase (LP) of the menstrual cycle were investigated. A total of 27 otherwise healthy patients previously scheduled for surgery were recruited after informed consent. An LH rise was detected, and CL were grouped according to age (Days 2-5 post-LH-rise, early LP; Days 6-10, mid LP; Days 11-14, late LP). Using a semiquantitative reverse transcription-polymerase chain reaction assay, the PR-B mRNA levels, which were 100- to 1000-fold lower than PR-A/B mRNA, were 46% lower (P < 0.05, n = 24) in mid LP, compared to early and late LP. CL tissue levels of progesterone and PR A/B protein levels were inversely correlated to increasing CL age; i.e., significantly reduced levels were observed in the late LP (r(2) = 0.34, P < 0.01, n = 23). Expression of PR-A/B mRNA as well as PR-A/B protein were detected by in situ hybridization and immunohistochemistry, respectively. Both methods revealed a clear and distinct localization to cells in the steroidogenic layer of the CL. Freshly obtained mid-luteal CL cells were cultured in vitro, and media were analyzed for progesterone concentrations after treatment by incremental doses of hCG and the stable PR antagonist mifepristone, alone or in combination. Mifepristone did not per se alter progesterone synthesis, but when it was added in conjunction with hCG, a dose-related inhibitory response was seen, with a maximal 47% reduction in progesterone output at a 10 microM addition (P < 0.05, n = 3). Collectively, these data implicate a stimulatory role of progesterone receptor-mediated action in the steroidogenic cells of the human CL, which may serve as an important pathway for maintaining functional homeostasis during early pregnancy. PMID- 10684808 TI - Developmental expression of thyroid hormone receptors in the rat testis. AB - Sertoli cell proliferation in the rat is completed by Days 15-20 postnatally. Thyroid hormones appear to regulate the duration of Sertoli cell proliferation, affecting adult Sertoli cell number and hence the capacity of the testis to produce sperm. In the present study, a combination of immunohistochemistry, immunoblot analysis, and reverse transcription-polymerase chain reaction was used to demonstrate the expression pattern of thyroid hormone receptors (TR) in the juvenile and adult rat testis. The results indicated that TRalpha1 was expressed in proliferating Sertoli cell nuclei, its expression decreasing coincident with the cessation of proliferation. TRalpha2, TRalpha3, and TRbeta1 mRNAs were expressed at low levels during development; however, the corresponding protein was not detected by immunoblot analysis. In addition, TRalpha1 was found to be expressed in germ cells from intermediate spermatogonia to mid-cycle pachytene spermatocytes. Immunohistochemistry also demonstrated TR expression in a subset of interstitial cells. The demonstration of TR expression in germ cells undergoing spermatogenic differentiation suggests a possible role for thyroid hormones in the adult testis. PMID- 10684809 TI - Expression of steroidogenic factor 1 in the testis requires an E box and CCAAT box in its promoter proximal region. AB - Steroidogenic factor 1 (SF-1), also known as adrenal 4-binding protein, is a member of the nuclear hormone receptor family that regulates transcription of genes encoding hormones and steroidogenic enzymes important to the function of the hypothalamic-pituitary-gonadal axis. The mammalian Ftz-F1 gene encodes SF-1 and is required for development of adrenal glands and gonads. To better understand the mechanisms regulating this gene in the gonads, we have examined its expression in the testis and characterized the promoter region for SF-1 in two testicular cell types. SF-1 promoter activity was examined in primary cultures of Sertoli cells and cell lines representative of Sertoli and Leydig cells. Deletion mutagenesis of the promoter identified several regions: both 5' and 3' to the transcriptional start sites that are important for transcriptional activity. Two elements, an E box and a CCAAT box, were found to be important for SF-1 transcription in the testis. An oligodeoxynucleotide containing both of these elements bound three specific protein complexes. The binding of one complex required only sequences within the E box and cross-reacted with antibodies against the basic helix-loop-helix ZIP proteins USF1 and USF2. A second specific complex required sequences within both the E box and CCAAT box for efficient binding, while a third complex predominantly interacted with sequences within the CCAAT motif. The presence of multiple protein complexes binding these sites suggests that regulation through these elements may involve interactions with different factors that depend on the state of the cell and its environment. PMID- 10684810 TI - Changes in the testis interstitium of Sprague Dawley rats from birth to sexual maturity. AB - Changes in the rat testis interstitium from birth to adulthood were studied using Sprague Dawley rats of 1, 7, 14, 21, 28, 40, 60, and 90 days of age. Our objectives were 1) to understand the fate of the fetal Leydig cells (FLC) in the postnatal rat testis, 2) to determine the volume changes in testicular interstitial components and testicular steroidogenic capacity in vitro with age, 3) to differentially quantify FLC, adult Leydig cells (ALC), and different connective tissue cell types by number and average volume, and 4) to investigate the relationship between mesenchymal and ALC numbers during testicular development. FLC were present in rat testes from birth to 90 days, and they were the only steroidogenic cells in the testis interstitium at Days 1 and 7. Except for FLC, all other interstitial cell numbers and volumes increased from birth to 90 days. The average volume of an FLC and the absolute volume of FLC per testis were similar at all ages except at Day 21, when lower values were observed for both parameters. FLC number per testis remained constant from birth through 90 days. The observations suggested that the significance of FLC in the neonatal prepubertal rat testis is to produce testosterone to activate the hypothalamo hypophyseal-testicular axis for the continued development of the male reproductive system. ALC were the abundant Leydig cell type by number and absolute volume per testis from Day 14 onwards. The absolute numbers of ALC and mesenchymal cells per testis increased linearly from birth to 90 days, with a slope ratio of 2:1, respectively, indicating that the rate of production of Leydig cells is 2-fold greater than that of mesenchymal cells in the postnatal rat testis through 90 days. In addition, this study showed that the mesenchymal cells are an active cell population during testis development and that their numbers do not decrease but increase with Leydig cell differentiation and testicular growth up to sexual maturity (90 days). PMID- 10684811 TI - Human endometrial endothelial cells: isolation, characterization, and inflammatory-mediated expression of tissue factor and type 1 plasminogen activator inhibitor. AB - Binding of Ulex europaeus lectin to microvessels was used to isolate endothelial cells from cycling human endometrium. Cultured human endometrial endothelial cells (HEECs) exhibited endothelial cell-specific characteristics such as tube formation on a basement membrane matrix and sequestration of acetylated low density lipoprotein. Markers for potentially contaminating epithelial, stromal, smooth muscle, and bone marrow-derived cells were not detected in the HEEC cultures. Basal and proinflammatory-stimulated immunostaining profiles for endothelial cell-specific adhesion markers, as exemplified by Von Willebrand's factor and E-selectin, were similar for cultured HEECs and human umbilical venous cord endothelial cells (HUVECs). However, HUVECs expressed several extracellular matrix proteins that were absent from cultured HEECs. In the latter, the protein kinase C agonist phorbol myristate acetate transiently enhanced tissue factor (TF) mRNA levels and elicited a more prolonged elevation in TF protein levels, but did not affect plasminogen activator inhibitor-1 (PAI-1) mRNA and protein levels. Inappropriate expression of TF, which initiates hemostasis by generating thrombin, and of PAI-1, which regulates hemostasis by acting as the primary inhibitor of fibrinolysis, can each lead to thrombosis. The differential regulation of TF and PAI-1 expression revealed in the current study emphasizes the importance of using HEECs to evaluate mechanisms regulating the hemostatic/thrombotic balance in human endometrium. PMID- 10684812 TI - Gestational profile of leptin messenger ribonucleic acid (mRNA) content in the placenta and adipose tissue in the rat, and regulation of the mRNA levels of the leptin receptor subtypes in the hypothalamus during pregnancy and lactation. AB - Serum leptin levels were significantly increased during rat gestation. Our data showed that leptin mRNA levels in both the adipose tissue and placenta were higher as pregnancy progressed, suggesting a role for both tissues in the hyperproduction of leptin. This paradoxical increase in leptin concentration during gestation suggests that a physiological state of leptin resistance may exist at the hypothalamic level that may explain the hyperphagia observed in pregnant rats. In order to study this issue further, levels of the mRNA encoding the different leptin receptor isoforms were determined in the hypothalamus of pregnant and nonpregnant rats. We found a specific reduction of the mRNA levels encoding the leptin receptor isoform Ob-Rb in the hypothalamus of pregnant rats compared to nonpregnant animals, suggesting that during pregnancy the hypothalamus shows a physiological resistance to the high levels of leptin due, at least in part, to a decrease in the expression of the long, biologically active form of the leptin receptor (Ob-Rb). During lactation, serum leptin levels returned to values observed in nonpregnant rats. In the hypothalami of these animals, Ob-Rb mRNA content was similar to that observed in nonpregnant rats, but we found an increased expression of some of the short forms of the leptin receptor (Ob-Re and Ob-Rf). This could contribute to induction of the hyperphagia present during lactation. These data provide new insights into the adaptive mechanisms that take place during pregnancy and lactation in order to meet increased metabolic requirements. PMID- 10684813 TI - Characterization of ovarian function in granulocyte-macrophage colony-stimulating factor-deficient mice. AB - During the estrous cycle and early pregnancy, lymphohemopoietic cytokines and chemokines contribute to the regulation of ovarian function by orchestrating the recruitment and activation of leukocytes associated with the ovulatory follicle and corpus luteum. The purpose of this study was to investigate the physiological role of granulocyte-macrophage colony-stimulating factor (GM-CSF) in the ovary, utilizing mice genetically deficient in GM-CSF. Our results show that the mean duration of the estrous cycle in GM-CSF-deficient (GM-/-) mice was extended by 1.5 days (mean +/- SE, 4.9 +/- 0.3 vs. 6.5 +/- 0.5 days for GM+/+ and GM-/- mice, respectively). Similar ovulation rates were observed in immature superovulated mice (31.8 +/- 7.7 vs. 28.9 +/- 6.4 oocytes per mouse) and adult naturally cycling mice (10.4 +/- 0.8 vs. 10.3 +/- 0.8 oocytes per mouse). Furthermore, comparable numbers of oocytes were released from GM+/+ and GM-/- ovaries in an in vitro perfusion model. However, ovaries in pregnant GM-/- mice were found to comprise fewer cells and synthesize less progesterone (141.6 +/- 10.3 vs. 116.5 +/- 6 nM plasma), although the duration of pseudopregnancy was unaltered by GM CSF deficiency (11.0 +/- 0.2 vs. 11.0 +/- 0.5 days). Immunohistochemical staining of leukocytes in the ovary during the periovulatory period indicated that the size and composition of ovarian leukocyte populations were unaltered in the absence of GM-CSF. However, an effect of GM-CSF deficiency on the activation phenotype of ovarian leukocytes was indicated by a 57% increase in mean secretion of nitric oxide in in vitro-perfused GM-/- ovaries, and diminished major histocompability complex (MHC) class II (Ia) expression in ovarian macrophages and/or dendritic cells (30.5 +/- 7. 2% vs. 9.1 +/- 1.8% positive stain in GM+/+ and GM-/- ovaries, respectively). Furthermore, ovarian macrophages and neutrophils were diminished in number after parturition, with significantly decreased CD11b+ (Mac-1) staining in the stromal region of postpartum GM-/- ovaries (6.7 +/- 0.6 vs. 3.6 +/- 0.7% positive stain). In summary, GM-CSF does not appear to be essential for ovarian function but may play a role in fine tuning the activation status and adhesive properties of ovarian myeloid leukocytes. Aberrant activation of these cells appears to compromise the luteinization process and the steroidogenic capacity of the corpus luteum during early pregnancy in GM-CSF-deficient mice. PMID- 10684814 TI - Ontogeny of angiotensin II type 1 receptor and cytochrome P450(c11) in the sheep adrenal gland. AB - In the present study we investigated the ontogeny of the expression of the type 1 angiotensin receptor (AT(1)R mRNA) and the zonal localization of AT(1)R immunoreactivity (AT(1)R-ir) and cytochrome P450(c11) (CYP11B-ir) in the sheep adrenal gland. In the adult sheep and in the fetus from as early as 90 days gestation, intense AT(1)R-ir was observed predominantly in the zona glomerulosa and to a lesser extent in the zona fasciculata, and it was not detectable in the adrenal medulla. AT(1)R mRNA decreased 4-fold between 105 days and 120 days, whereas AT(1)R mRNA levels remained relatively constant between 120 days and the newborn period. In contrast, both in the adult sheep and in the fetal sheep from as early as 90 days gestation, intense CYP11B-ir was consistently detected throughout the adrenal cortex and in steroidogenic cells that surround the central adrenal vein. In conclusion, we speculate that the presence of AT(1)R in the zona fasciculata, and the higher levels of expression of AT(1)R at around 100 days gestation, may suggest that suppression of CYP17 is mediated via AT(1)R at this time. The abundant expression of AT(1)R-ir and CYP11B-ir in the zona glomerulosa of the fetal sheep adrenal gland would also suggest that lack of angiotensin II stimulation of aldosterone secretion is not due to an absence of AT(1)R or CYP11B in the zona glomerulosa. PMID- 10684815 TI - Mouse testis brain ribonucleic acid-binding protein/translin colocalizes with microtubules and is immunoprecipitated with messenger ribonucleic acids encoding myelin basic protein, alpha calmodulin kinase II, and protamines 1 and 2. AB - Testis brain RNA-binding protein (TB-RBP) is a sequence-dependent RNA-binding protein that binds to conserved Y and H sequence elements present in many brain and testis mRNAs. Using recombinant TB-RBP and a highly enriched tubulin fraction, we demonstrate here that recombinant TB-RBP binds to microtubules assembled in vitro. The interaction between recombinant TB-RBP and microtubules was inhibited by high salt and by the microtubule disassembling agents colcemid and calcium, but not by the microfilament-disassembling agent cytochalasin D. Confocal microscopy confirmed colocalization of TB-RBP and tubulin in the cytoplasm of male germ cells. An affinity-purified antibody prepared against recombinant TB-RBP specifically precipitated mRNAs encoding myelin basic protein and alpha calmodulin-dependent kinase II-two transported mRNAs, and protamines 1 and 2-two translationally regulated testicular mRNAs. These data indicate an intracellular association between TB-RBP and specific target mRNAs and suggest an involvement of TB-RBP in microtubule-dependent mRNA transport in the cytoplasm of cells. PMID- 10684816 TI - In vivo oxytocin release from microdialyzed bovine corpora lutea during spontaneous and prostaglandin-induced regression. AB - The release of luteal oxytocin during spontaneous and prostaglandin-induced luteolysis was investigated in cows. A continuous-flow microdialysis system was used in 11 cows to collect dialysates of the luteal extracellular space between Days 12 and 24 postestrus. Seven cows were untreated and were expected to exhibit spontaneous luteolysis during sampling, whereas 4 cows received prostaglandin F(2alpha) (PGF(2alpha)) systemically between Days 13 and 15 to induce luteolysis during sampling. Oxytocin was detectable in the dialysate of all cows before Day 16 postestrus and occurred as 2 or 3 discrete pulses per 12-h sampling period. For non-PGF(2alpha)-treated cows, dialysate oxytocin content began to decline spontaneously on Day 15 postestrus and was undetectable by Day 17 postestrus. Oxytocin decay curves preceded onset of serum progesterone decline by at least 72 h and were not related temporally with onset of progesterone decline within cow. Exogenous PGF(2alpha) (25 mg, i.m.) produced a 10-fold increase in dialysate oxytocin within 1 h (1.9 +/- 0.3 pg/ml to 20.8 +/- 3.0 pg/ml; P < 0. 01). Dialysate oxytocin then declined to pretreatment concentrations within 2 h and was undetectable within 8 h posttreatment. A second PGF(2alpha) injection given 20 h after the first did not result in a measurable increase in dialysate oxytocin, probably because luteolysis was underway. Although robust luteal oxytocin release was observed after treatment with a pharmacological dose of PGF(2alpha), the lack of detectable oxytocin secretion during spontaneous luteolysis suggests that the contribution of luteal oxytocin in the cow may be less than that proposed for the ewe. PMID- 10684817 TI - Inhibitory and stimulatory interactions between endogenous gonadotropin-releasing hormones in the African catfish (Clarias gariepinus). AB - In the brain of all vertebrate classes, chicken (c) GnRH-II ([His(5), Trp(7),Tyr(8)]GnRH, cGnRH-II) is expressed in the mesencephalon. In addition, at least one other form of GnRH is expressed in the preoptical area/hypothalamus. In the human pituitary stalk and the mouse median eminence, cGnRH-II is present together with mammalian GnRH. Similarly, in the pituitary of several teleost fish (e.g., goldfish and eel, but not salmon or trout), a teleost GnRH is found together with cGnRH-II. These GnRHs are not colocalized in the same cells. Hence, these GnRH peptides may differentially regulate gonadotropin secretion and, in addition, may exert their effects simultaneously. The current study therefore investigated the effects of combinations of the two forms of GnRH present in the African catfish (Clarias gariepinus) pituitary-cGnRH-II and catfish GnRH ([His(5),Asn(8)]GnRH, cfGnRH)-on the cytosolic free calcium concentration ([Ca(2+)](i)) in single, Fura-2-loaded catfish gonadotrophs, as well as their effects on both in vitro and in vivo LH secretion. Both inhibitory and stimulatory effects of combinations of cfGnRH and cGnRH-II on [Ca(2+)](i) were observed, which were mirrored by their effects on both in vitro and in vivo LH secretion. The following pattern became apparent. The effect of intermediate or maximal effective cfGnRH doses was inhibited by the simultaneous presence of subthreshold or borderline effective cGnRH-II doses. Conversely, subthreshold or borderline effective concentrations of cfGnRH enhanced the effects of intermediate and maximal concentrations of cGnRH-II. In addition, combinations of cfGnRH and cGnRH-II concentrations that were equally active when tested separately showed an additive effect. The observed interactions between the two GnRHs may be of particular physiological relevance in the control of seasonal LH levels in the African catfish, as well as in other teleost species. Moreover, the occurrence of mutual inhibitory and stimulatory interactions between endogenous GnRHs may be a widespread aspect of GnRH action in vertebrates. PMID- 10684818 TI - Extracellular matrix composition and hypoxia regulate the expression of HLA-G and integrins in a human trophoblast cell line. AB - During human placentation, extravillous cytotrophoblast cells emerge from chorionic villi contacting the decidua to invade the uterine wall. When isolated from first-trimester placentae, cytotrophoblast cells undergo step-wise differentiation in vitro that recapitulates the phenotypic heterogeneity observed in vivo. We examined a cell line, HTR-8/SVneo, that has been established from human first-trimester cytotrophoblast to determine whether these cells possess some of the unique cytotrophoblast characteristics that have been described previously. Exposure during serum-free culture to hypoxic conditions (2% oxygen concentration) increased HTR-8/SVneo cell proliferation and reduced invasion of a three-dimensional basement membrane (Matrigel). During culture on surfaces coated with individual extracellular matrix proteins, HTR-8/SVneo cells expressed cytokeratin but not the trophoblast-specific major histocompatibility protein, HLA-G. However, HLA-G expression was induced in HTR-8/SVneo cells that contacted Matrigel. Expression of the alpha5 integrin subunit was relatively unaffected by matrix composition, whereas alpha1 was up-regulated and alpha6 was down-regulated after transferring cells to Matrigel. Hypoxia increased alpha6 and decreased both alpha1 and HLA-G expression on Matrigel. HTR-8/SVneo cells retain several important characteristics associated with primary cultures of first-trimester human cytotrophoblast cells, including their altered behavior in response to a changing maternal environment. PMID- 10684819 TI - Connexin gene expression in seminiferous tubules of the Sprague-Dawley rat. AB - Sertoli and spermatogenic cells establish germ cell- and epithelial stage dependent networks of cell-cell communication thought to be important for the initiation and maintenance of spermatogenesis. Since gap junctions assemble between Sertoli cells and between Sertoli and spermatogenic cells, it was hypothesized that multiple, unique routes of cell-cell communication may be established, in part, by the assembly of structurally diverse gap junctions from the connexin (Cx) multigene family. Differences in channel structure may support differences in ion or second messenger permeability between cell types. Reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that 11 Cx mRNAs were present in total RNA from seminiferous tubules and that 10 of the Cx mRNAs were present in polysomes and presumably translated. RT-PCR analyses also showed that the Cx mRNA population varied between different seminiferous tubule cell types. There were 9 Cx mRNAs in germ cells, 8 in Sertoli cells, and 5 in peritubular cells. One of the Cx mRNAs, Cx-50, was detected only in pachytene spermatocytes and round spermatids. Comparisons of the Cx mRNAs present in tubules at different postnatal ages showed that at least 2 Cxs (Cx-33 and Cx-50) accumulated when leptotene-zygotene stages developed. The multiple Cx genes and proteins produced in spermatogenesis may support the assembly of structurally diverse gap junctions. PMID- 10684820 TI - Polarized distribution of NHE1 and NHE2 in the rat epididymis. AB - Previous studies from our laboratory have provided evidence that the rat epididymis utilizes the Na(+)/H(+) exchanger to transport acid and base. The present study was undertaken to use immunohistochemistry for investigating the localization (apical versus basolateral) and distribution of NHE1 and NHE2 proteins along intact rat epididymis. Both proteins were found to be exclusively localized within the epithelium. Immunoreactivity for NHE1 was detected on the basolateral surface, whereas NHE2 immunoreactivity was detected on the apical side of the epithelium. Interestingly, NHE1 was found along the entire length of the epididymal tubule whereas NHE2 was absent in the initial segment but present in the caput, corpus, and cauda regions. These results, when interpreted along with those of previous functional studies, may suggest that the apical NHE2 is involved in Na(+) reabsorption and the basolateral NHE1 in HCO(3)(-) secretion in the rat epididymis. PMID- 10684821 TI - Evidence for progesterone receptors in the human fetoplacental vascular tree. AB - The presence of progesterone receptors (PR) throughout the human term fetoplacental vascular tree was investigated. By reverse transcription-polymerase chain reaction (RT-PCR), we showed expression of PR mRNAs in stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins. Binding studies and Scatchard analysis revealed a single class of high-affinity binding sites for (3)H-R5020 (promegestone) in cytosolic extracts of all placental vessels, with K(d) values in the range of 2.5-4 nM. High levels of PR were detected in placental vessels compared to other vascular tissues. Thus, maximum binding capacities of stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins were 247 +/- 25, 377 +/- 58, 295 +/- 40, 371 +/- 118, and 672 +/- 144 fmol/mg protein, respectively. Endothelial cell elimination in chorionic arteries did not significantly modify the number of PR. RT-PCR and binding studies also assessed PR expression in cultured placental vascular smooth muscle cells isolated from stem villi vessels. All these data suggested that most of the PR of fetoplacental vessels were from the media. In conclusion, we report here the first evidence of the presence of PR in the muscular layer of human term fetoplacental vessels. This finding, together with the high progesterone concentrations in cord blood, suggests that the interactions between the PR and its ligand may play a role in the physiology and physiopathology of human fetoplacental vascularization. PMID- 10684822 TI - Independent and hetero-oligomeric-dependent sperm binding to egg envelope glycoprotein ZPC in Xenopus laevis. AB - Vitelline envelopes are composed of glycoproteins that participate in sperm-egg interactions during the initial stages of fertilization. In Xenopus laevis, the vitelline envelope is composed of at least 4 glycoproteins (ZPA, ZPB, ZPC, and ZPX). A sperm binding assay involving the covalent coupling of envelope glycoproteins to silanized glass slides was developed. In our assay, sperm bound to the egg envelopes derived from oviposited eggs but not activated eggs. The majority of the egg envelope ligand activity for sperm binding was derived from the complex N-linked oligosaccharides of ZPC. This sperm binding involved N acetylglucosamine and fucose residues, as binding was abolished after treatment with cortical granule beta-N-acetylglucosaminidase and commercial beta-N acetylglucosaminidases and was reduced by 44% after treatment with alpha fucosidase. Although both the envelope glycoproteins ZPA and ZPC possessed independent ligand activity, ZPC was the major ligand for sperm binding (75%). Mixing of isolated ZPA, ZPB, and ZPC in a ratio of 1:4:4 (equal to that in the egg envelope) resulted in sperm binding that was greater than that of the sum of the separate components. The egg glycoproteins acted in synergy to increase sperm binding. Thus, ZPC possessed both independent and hetero-oligomeric-dependent ligand activities for sperm binding. PMID- 10684823 TI - Meiosis-activating sterol-mediated resumption of meiosis in mouse oocytes in vitro is influenced by protein synthesis inhibition and cholera toxin. AB - To explore the possible signaling pathways of meiosis-activating sterol (MAS) induced oocyte maturation and to elucidate whether the MAS pathway involves transcription or translation, arrested immature mouse oocytes were cultured with either the protein synthesis inhibitor cycloheximide or the heteronuclear RNA inhibitors alpha-amanitin or actinomycin D, respectively. Moreover, the possible involvement of a G protein-coupled receptor mechanism in MAS-mediated oocyte maturation was explored by influencing oocyte maturation with cholera toxin (CT). MAS-induced oocyte maturation was completely blocked by the addition of 50 microg/ml cycloheximide 4 h before the addition of MAS. Simultaneous addition of MAS and the protein synthesis inhibitor also significantly reduced the meiotic resumption compared to that in MAS-treated controls. In contrast, neither of the treatment regimens to inhibit transcription of DNA to RNA was observed to have any effect on the MAS-induced resumption of meiosis. CT was observed to inhibit MAS-induced, but not spontaneous, oocyte maturation in vitro, suggesting a putative involvement of G protein-coupled receptor mechanism in the MAS mode of action. In conclusion, protein synthesis was found to be an essential requirement for maintaining the oocytes' responsiveness to MAS-induced resumption of meiosis, in contrast to transcription. PMID- 10684824 TI - Endothelial vasodilator production by uterine and systemic arteries. IV. Cyclooxygenase isoform expression during the ovarian cycle and pregnancy in sheep. AB - Uterine artery endothelial production of the potent vasodilator, prostacyclin, is greater in pregnant versus nonpregnant sheep and in whole uterine artery from intact versus ovariectomized ewes. We hypothesized that uterine artery cyclooxygenase (COX)-1 and/or COX-2 expression would be elevated during pregnancy (high estrogen and progesterone) and the follicular phase of the ovarian cycle (high estrogen/low progesterone) as compared to that in luteal phase (low estrogen/high progesterone) or in ovariectomized (low estrogen and progesterone) ewes. Uterine and systemic (omental) arteries were obtained from nonpregnant luteal-phase (LUT; n = 10), follicular-phase (FOL; n = 11), and ovariectomized (OVEX; n = 10) sheep, as well as from pregnant sheep (110-130 days gestation; term = 145 +/- 3 days; n = 12). Endothelial and vascular smooth muscle (VSM) COX 1 protein levels and uterine artery endothelial cell COX-1 mRNA levels were compared. Using immunohistochemistry and Western analysis, the primary location of COX-1 protein was the endothelium; that is, we observed 2.2-fold higher COX-1 protein levels in intact versus endothelium-denuded uterine artery and a 6.1-fold higher expression in the endothelium versus VSM (P < 0.05). COX-2 protein expression was not detectable in either uterine artery endothelium or VSM. COX-1 protein levels were observed to be higher (1.5-fold those of LUT) in uterine artery endothelium from FOL versus either OVEX or LUT nonpregnant ewes (P < 0.05), with substantially higher COX-1 levels seen in pregnancy (4.8-fold those of LUT). Increases in uterine artery endothelial COX-1 protein were highly correlated to increases in the level of COX-1 mRNA (r(2) = 0.66; P < 0.01) for all treatment groups (n = 6-8 per group), suggesting that increased COX-1 protein levels are regulated at the level of increased COX-1 mRNA. No change in COX-1 expression was observed between groups in a systemic (omental) artery. In conclusion, COX-1 expression is specifically up-regulated in the uterine artery endothelium during high uterine blood flow states such as the follicular phase and, in particular, pregnancy. PMID- 10684825 TI - Expression of deoxyribonucleic acid repair enzymes during spermatogenesis in mice. AB - Meiotic recombination during gametogenesis is critical for proper chromosome segregation. However, the participating proteins and mechanics of recombination are not well understood in mammals. DNA repair enzymes play an essential role in both mitosis and meiosis in yeast. The mammalian mismatch repair system consists of homologues of the bacterial MutH, MutL, and MutS proteins. As part of our goal of understanding the function of enzymes that mediate meiotic recombination, we used a reverse transcription-polymerase chain reaction approach to identify germ cell transcripts for the MutL homologue, Pms2, and two members of the MutS family, Msh2 and Msh3. Both the Pms2 and the Msh2 genes were highly expressed in mitotically proliferating spermatogonia, and early in meiotic prophase in the leptotene and zygotene spermatocytes. Thereafter, expression declined in early and mid pachytene spermatocytes, and was negligible in postmeiotic spermatids. In contrast, expression of Msh3 was at its highest level in pachytene spermatocytes. Protein levels were similar to gene expression patterns, and both PMS2 and MSH2 were localized in spermatogonia and spermatocytes. These patterns of expression for genes encoding mismatch repair enzymes are consistent with the proposed roles of the gene products in mismatch repair during both DNA replication and recombination. PMID- 10684826 TI - Fetal-to-maternal progression of prostaglandin H(2) synthase-2 expression in ovine intrauterine tissues during the course of labor. AB - We examined whether spontaneous parturition in sheep was associated with tissue specific changes in prostaglandin H(2) synthase-2 (PGHS-2) expression and/or with altered expression of myometrial EP and FP receptors. Placental and uterine tissues were collected from three groups of chronically catheterized sheep in relation to term spontaneous labor: late pregnancy, not in labor; early labor; and active labor. Expression of PGHS-2 mRNA and protein was determined by in situ hybridization, Western blotting, and immunohistochemistry. Semiquantitative reverse transcription-polymerase chain reaction was used to assess the presence of and changes in prostaglandin (PG) receptor subtypes. In placenta, PGHS-2 mRNA and protein localized to trophoblast uninucleate cells and tended to increase with early labor. PGHS-2 mRNA and protein localized to endometrial epithelium and to myometrium, where PGHS-2 protein levels rose in active labor tissues. Concentrations of PGE(2) in fetal plasma rose progressively with labor, whereas 13,14-dihydro-15-keto-PGF(2alpha) in maternal plasma increased significantly only in active labor. Messenger RNA encoding four EP receptor subtypes and FP receptor were present in myometrium, but levels did not change with labor. We suggest that spontaneous labor in sheep is associated with a progressive increase in PGHS-2 expression in a temporal and tissue-specific manner from trophoblast to maternal tissues, rather than alteration in PG receptor gene expression. PMID- 10684827 TI - Testosterone-dependent primer pheromone production in the sebaceous gland of male goat. AB - To test the hypothesis that the primer pheromone responsible for inducing the "male effect" is produced in the sebaceous gland androgen dependently, we examined the correlation between morphological changes of sebaceous glands and the pheromone activity in skin samples taken from castrated goats that had been treated with testosterone. Five castrated goats were implanted s.c. with testosterone capsules to maintain physiological levels of plasma testosterone for four weeks. Skin samples were obtained from the head region on Day 0 (the day of testosterone implant), Day 7, Day 14, Day 28 (the day of testosterone removal), Day 36, Day 42, and Day 56. Matched blood samples were also collected for measurement of testosterone concentration. The pheromone activity of the ether extracts of the upper dermal layer containing sebaceous glands was assessed by its stimulatory effect on the hypothalamic GnRH pulse generator, which was monitored for changes of specific multiple unit activity (MUA) in ovariectomized estradiol-primed goats as described previously. The sebaceous gland enlarged during the testosterone treatment but reduced in size after testosterone removal. The pheromone activity first appeared in 2 out of 5 goats on Day 7 and in all the 5 goats by Day 28. Fourteen days after testosterone removal (Day 42), the pheromone activity was no longer detectable in any of the 5 goats. In short, the sebaceous gland size and the pheromone activity shifted almost in parallel. The present results provide strong support for the view that the primer pheromone is produced testosterone dependently in the sebaceous gland of the male goat. PMID- 10684828 TI - Involvement of protein kinase A and A kinase anchoring protein in the progesterone-initiated human sperm acrosome reaction. AB - The signal transduction pathways involved in the progesterone (P(4))-initiated mammalian sperm acrosome reaction (AR) are not fully understood. To investigate the role of the protein kinase A (PKA) pathway in the P(4)-initiated AR, we probed this pathway by pretreating capacitated human sperm with reagents designed to either inhibit PKA activation or disrupt PKA/A kinase anchoring protein (AKAP) interactions. Preincubation with the stearated (membrane permeable) PKA inhibitor, PKI alpha 5-24 (S-PKI alpha 5-24), significantly inhibited the P(4) initiated AR at 10 microM as compared to stearated control peptide. In contrast, preincubation with 100 microM nonstearated PKI alpha 5-24 did not significantly inhibit versus solvent control. Preincubation with the PKA inhibitor Rp-8-Br-cAMP at 500 microM and 150 microM significantly inhibited the P(4)-initiated AR versus 8-Br-cAMP and versus solvent. Preincubation with the anchoring inhibitory peptide S-Ht-31 significantly stimulated the P(4)-initiated AR at 10, 3, and 1 microM versus inactive control peptide. The stimulation of the P(4)-initiated AR by 3 microM S-Ht31 was significantly inhibited by the addition of 30 microM S-PKI alpha 5-24 prior to the addition of S-Ht31. Preincubation with S-PKI alpha 5-24 (30 microM) partially inhibited the ionomycin (50 microM)-initiated AR. A role for PKA in the P(4)-initiated AR may exist both upstream and downstream of Ca(2+) entry. Our studies present the first evidence for the participation of PKA in the P(4)-initiated AR and also suggest that AKAPs are involved in the PKA-mediated events. PMID- 10684829 TI - Uveitis in HIV positive patients. PMID- 10684830 TI - Open invitation from the International Poverty and Health Network to all health professionals. PMID- 10684831 TI - Clinicopathological correlation in exudative age related macular degeneration: histological differentiation between classic and occult choroidal neovascularisation. AB - AIMS: To analyse the histopathology of classic and occult choroidal neovascular membrane surgical specimens in age related macular degeneration. METHODS: 35 membranes, from a consecutive series of surgically removed choroidal neovascular membranes in age related macular degeneration, were classified as classic or occult following the guidelines of the Macular Photocoagulation Study. Membranes with classic as well as occult components were considered as mixed membranes. The membranes were serially sectioned and stained with haematoxylin and eosin, Masson trichrome, periodic acid-Schiff, and phosphotungstic acid haematoxylin stain. The correlation has been made in a masked fashion. RESULTS: 31 membranes (19 classic, 10 occult, and two mixed membranes) could be analysed histologically. 18 classic choroidal neovascular membranes had a major subretinal fibrovascular component and 10 of these had an additional, minor fibrovascular component under the retinal pigment epithelium. The 10 occult membranes contained a fibrovascular component under the retinal pigment epithelium and the two mixed membranes contained fibrovascular tissue on both sides of the retinal pigment epithelium. Fibrin and remains of outer segments tended to occur at the lateral edges of classic membranes and to cover the inner surface of occult membranes. CONCLUSION: Classic choroidal neovascularisation in age related macular degeneration is predominantly composed of subretinal fibrovascular tissue while occult choroidal neovascularisation is composed of fibrovascular tissue at the choroidal side of the retinal pigment epithelium. PMID- 10684832 TI - Macular degeneration: do conventional measurements of impaired visual function equate with visual disability? AB - AIMS: To examine the relation between measures of vision and ability to perform daily living tasks in those visually impaired with macular degeneration. METHODS: A visual functioning index (daily living tasks dependent on vision: DLTV) was used to evaluate patients' perception of their ability to perform vision dependent tasks. Distance visual acuity, near visual acuity, reading speed, and contrast sensitivity were measured in all patients. In addition, a new measure of reading ability was derived, designated the reading index. This takes into account both the size of the text read and the time to read it and is equivalent to the reading speed in words per minute divided by text size in M. RESULTS: The reading index was found to show best associations with the majority of items within the DLTV. Stepwise regression identified the combination of reading index and distance visual acuity as having the best associations with DLTV items. The present study also demonstrated that specific levels of vision as measured by acuity, reading index, and contrast sensitivity corresponded with different perceived amounts of difficulty in the performance of daily living tasks. CONCLUSIONS: This study showed that reading index is valuable in predicting the ability to perform daily living tasks and therefore may be useful in the visual assessment of the visually impaired individual. In addition, this study identified specific levels of vision at which individuals reported different degrees of difficulty in performing daily living tasks. PMID- 10684833 TI - Radiological and clinicopathological features of orbital xanthogranuloma. AB - BACKGROUND: Orbital xanthogranuloma, a diagnosis confirmed histologically, occurs rarely in adults and children. With its characteristic macroscopic appearance the adult form may be associated with a spectrum of biochemical and haematological abnormalities including lymphoproliferative malignancies. METHOD: The clinicopathological features and imaging appearances on computed tomography and magnetic resonance imaging of this condition are described in eight adults and a child. RESULTS: Radiological evidence of proptosis was present in seven patients. In all nine patients an abnormal infiltrative soft tissue mass was seen, with increased fat in six cases. All patients had associated enlargement of extraocular muscles suggestive of infiltration and five had lacrimal gland involvement. Encasement of the optic nerve, bone destruction, and intracranial extension was present only in the child with juvenile xanthogranuloma. Haematological and/or biochemical abnormalities were detected in seven patients and seven patients had other systemic diseases which were considered to have an immune basis. One patient subsequently developed non-Hodgkin's lymphoma. CONCLUSION: The investigation and management of orbital xanthogranulomas requires a multidisciplinary approach even though the diagnosis may be suspected clinically. Imaging delineates the extent of disease and involvement of local structures and may influence the differential diagnosis. The juvenile form may be more locally aggressive, causing bone destruction with consequent intracranial extension. PMID- 10684834 TI - Prospective surveillance of sympathetic ophthalmia in the UK and Republic of Ireland. AB - AIMS: To establish current epidemiological data, risks, and interventional outcomes of newly diagnosed sympathetic ophthalmia (SO). METHODS: Prospective surveillance took place of all permanently employed ophthalmologists in the UK and Republic of Ireland by a monthly reporting card through the British Ophthalmological Surveillance Unit. Case ascertainment was made of newly diagnosed SO from July 1997 and questionnaire data were returned at baseline, 6 months, and 1 year after diagnosis. RESULTS: 23 patients with newly diagnosed SO were recruited over 15 months, corresponding to a minimum estimated incidence of 0.03/100 000. Baseline data were available on 18 patients, in whom SO occurred after surgery in 11 patients, after retinal surgery alone in six patients, and after accidental trauma in seven patients. 12 of the 16 patients with 1 year follow up had a visual acuity of 6/12 or better. Good visual outcome was related to prompt and adequate systemic immunosuppressive therapy. CONCLUSIONS: The incidence of sympathetic ophthalmia is very low. The main current risk is surgery, particularly retinal surgery, but visual prognosis is good if early diagnosis is made and rapid, adequate immunotherapy is commenced. PMID- 10684835 TI - Scanning laser polarimetry in normal subjects and patients with myopia. AB - AIMS: To examine the changes in the retinal nerve fibre layer (NFL) thickness with age and myopia in normal population. METHODS: Retinal nerve fibre layer thickness was measured with a scanning laser polarimeter (NFA-I) in 180 normal subjects of varying age (range 7-83 years) and in 110 eyes of 85 patients with myopia of varying degrees (range -1.00 to -15.00D). They were all voluntary Anatolian people. Superior to nasal (S/N), inferior to nasal (I/N), and the superior to inferior (S/I) ratios were used for the assessment of retinal NFL thickness. RESULTS: The mean superior NFL ratio was 2.96 and the mean inferior NFL ratio was 2.93 in normal subjects. There was a gradual decrease in NFL ratio with increasing age (simple regression analysis, p<0.05). The mean S/I ratio was 1.01 with a large variation. In patients with myopia, the mean superior NFL ratio was 2.60 and the mean inferior NFL ratio was 2.72. Superior and inferior NFL retardations, and S/I ratio in myopic patients were significantly (15.5%, 10.8%, and 4.9% respectively) lower than that of age matched normals (t test, p<0.05). There was also a gradual decrease in NFL thickness with increasing degree of myopia (simple regression analysis, p<0.05). CONCLUSIONS: Nomograms we obtained for retinal NFL thickness may serve as reference points for the assessment of normal Anatolian people and myopic patients in future studies. NFL thicknesses gradually decreased with increasing age. Patients with myopia had significantly lower NFL thicknesses than normal subjects and, although weakened by wide age range of myopic group, there is a linear relation between severity of myopia and NFL thickness in myopic patients. PMID- 10684836 TI - Treatment of conjunctival squamous cell carcinoma with topical 5-fluorouracil. AB - AIM: To evaluate the efficacy of topical 5-fluorouracil (5-FU) alone, without concurrent surgery or radiotherapy, for the treatment of conjunctival squamous cell carcinoma. METHODS: Eight patients affected by conjunctival squamous cell carcinoma (three recurrent cases, three incompletely excised, and two untreated cases) were treated with 1% 5-FU eye drops. Topical 1% 5-FU was administered four times daily for 4 weeks (one course). Clinical examination (biomicroscopy and photography) and morphological evaluation of conjunctival cytological specimens were used to monitor the efficacy of local chemotherapy, side effects, and recurrences. RESULTS: All patients showed clinical regression of conjunctival carcinoma after topical 1% 5-FU treatment. Neoplastic conjunctiva was completely replaced by normal epithelium within 3 months. Mean follow up was 27 months. One patient needed two courses of local chemotherapy for recurrent disease. An acute transient toxic keratoconjunctivitis was observed in all treated cases; it was easily controlled with topical therapy. No long term side effects were found. CONCLUSIONS: Topical 1% 5-FU is effective in the treatment of recurrent, incompletely excised, and selected untreated conjunctival squamous cell carcinomas. Topical 1% 5-FU has no major complications. This study suggests that topical conjunctival chemotherapy with 1% 5-FU may be useful, at least as adjunctive therapy, in the treatment of conjunctival squamous cell carcinoma. PMID- 10684837 TI - Autologous limbal transplantation in patients with unilateral corneal stem cell deficiency. AB - AIM: To describe a surgical technique for autologous limbal stem cell transplantation and the outcome of a series of patients with unilateral stem cell deficiency. METHODS: A report of six consecutive patients who underwent autologous limbal stem cell transplantation is presented. The primary diagnosis included alkali burn (n=3), conjunctival intraepithelial neoplasia (CIN) (n=1), recurrent pterygium (n=1), and contact lens induced keratopathy (n=1). The autologous transplanted tissue consisted of peripheral cornea, limbus, and conjunctiva obtained from the contralateral eye. Three of the above patients underwent penetrating keratoplasty in association with auto-limbal transplantation. A significant modification to established techniques was the close monitoring of conjunctival epithelial migration in the immediate postoperative period. If conjunctival epithelium threatened to migrate on to the corneal surface, it was mechanically removed at the slit lamp and prevented from crossing the limbus. This was required in three patients. RESULTS: The mean follow up was 18.8 months. The outcome was satisfactory in all cases: a stable corneal surface was restored and there was a substantial improvement in vision and symptoms. One patient had a primary failure of the corneal allograft associated with glaucoma, and 6 months later developed a retinal detachment. No complications were noted in the donor eye with the exception of one patient who developed filamentary keratitis along the edge of the donor site. CONCLUSION: Autologous limbal transplantation with corneal, limbal, and conjunctival carriers was found to be useful for ocular surface reconstruction, over a mid-term follow up, in patients with unilateral stem cell deficiency. Close monitoring of the migration of conjunctival epithelium in the immediate postoperative period, and preventing it from crossing the limbus, ensured that the corneal surface was re epithelialised exclusively from epithelial cells derived from the transplanted limbal tissue. This approach should improve the success of this procedure. PMID- 10684838 TI - Long term results after autologous nasal mucosal transplantation in severe mucus deficiency syndromes. AB - AIM: Severe mucus deficiency syndromes may require substitution of mucous membrane for re-establishment of the ocular surfaces. The long term results after autologous nasal mucosal transplantation were investigated. METHODS: 55 eyes of 50 patients with severe mucus deficiency syndromes were followed retrospectively after free autologous nasal mucosal transplantation-group A: patients after severe lye, acid, heat burns, or radiation (n=38 eyes), group B: patients with systemic mucosal disease (n=17 eyes). The results of routine clinical examination were recorded and patients were followed for a median of 37 months. 17 biopsies of transplanted nasal mucosa were studied by light microscopy and 22 patients by impression cytology before and at several intervals after mucosal transplantation. RESULTS: All nasal mucosal grafts healed well and no intraoperative complications occurred. During follow up 107 additional surgical procedures were performed including 16 lamellar and 21 penetrating keratoplasties. Subjective complaints improved in 44/47 patients with preoperative symptoms. Best corrected visual acuity at the end of follow up was increased in 23 eyes, 10 eyes (18. 2%) reached a final visual acuity equal to or greater than 20/200. Histopathologically, all (n=17) biopsies showed vital intraepithelial mucin producing goblet cells in the nasal mucosal graft (median 25 cells/field (400x magnification)). The mean density of goblet cells before transplantation was 48/mm(2) and after nasal mucosal grafting 432/mm(2) measured by impression cytology (p<0. 0001). CONCLUSIONS: Functional goblet cells persist in autologous nasal mucosa for up to 10 years after transplantation. In patients with severe mucus deficiency syndromes of different origin nasal mucosal transplantation can re-establish the ocular surface, substitute the mucus components of the tear film, improve symptoms of the patients, and facilitate a moderate increase in visual acuity. PMID- 10684839 TI - Clinical course of hurricane keratopathy. AB - BACKGROUND/AIMS: "Hurricane keratopathy" is the name given to the whorl pattern, highlighted with fluorescein, seen in situations where corneal epithelial cell turnover is exaggerated. Although the condition is well described, follow up data on patients with this condition and its sequelae have only been reported in corneal graft patients. The aim was to study the clinical course of hurricane keratopathy in corneal graft patients and contact lens wearers, and to document any sequelae of this condition. METHODS: Hurricane keratopathy, occurring in 20 eyes with corneal grafts and 16 eyes (six bilateral) wearing rigid gas permeable contact lenses, was studied and followed. The occurrence, pattern, progress, resolution, and residual effects of the whorls were noted. RESULTS: Hurricane keratopathy was noted to occur in grafts as previously reported and also in contact lens wearers, which has hitherto not been reported. The whorls usually appeared within the first 3 weeks postoperatively and persisted up to 4 months. A small epithelial defect (11.1%), heaped epithelial cells (5.6%), and a nebular grade opacity (2.8%), were the only significant sequelae noted at the epicentre of the whorls. Resolution occurred from the periphery towards the centre of the cornea. CONCLUSIONS: The whorl pattern is sustained as long as the stimulus for increased cell turnover is maintained. Once this stimulus is eliminated, the pattern tends to resolve spontaneously. PMID- 10684840 TI - Epidemiology of pterygium in Victoria, Australia. AB - AIM: To describe the prevalence of and risk factors for pterygium in a population based sample of residents of the Australian state of Victoria who were aged 40 years and older. METHODS: The strata comprised nine randomly selected clusters from the Melbourne statistical division, 14 nursing homes randomly selected from the nursing homes within a 5 kilometre radius of the nine Melbourne clusters, and four randomly selected clusters from rural Victoria. Pterygium was measured in millimetres from the tip to the middle of the base. During an interview, people were queried about previous ocular surgery, including surgical removal of pterygium, and their lifetime exposure to sunlight. RESULTS: 5147 people participated. They ranged in age from 40 to 101 years and 2850 (55.4%) were female. Only one person in the Melbourne cohort reported previous pterygium surgery, and seven rural residents reported previous surgery; this information was unavailable for the nursing home residents. Pterygium was present upon clinical examination in 39 (1.2%) of the 3229 Melbourne residents who had the clinical examination, six (1. 7%) of the nursing home residents, and 96 (6.7%) of the rural residents. The overall weighted population rate in the population was 2.83% (95% CL 2.35, 3.31). The independent risk factors for pterygium were found to be age (OR=1.23, 95% CL=1.06, 1.44), male sex (OR=2.02, 95% CL=1.35, 3.03), rural residence (OR=5.28, 95% CL=3. 56, 7.84), and lifetime ocular sun exposure (OR=1.63, 95% CL=1.18, 2. 25). The attributable risk of sunlight and pterygium was 43.6% (95% CL=42.7, 44.6). The result was the same when ocular UV-B exposure was substituted in the model for broad band sun exposure. CONCLUSION: Pterygium is a significant public health problem in rural areas, primarily as a result of ocular sun exposure. PMID- 10684841 TI - Effects of topical nipradilol, a beta blocking agent with alpha blocking and nitroglycerin-like activities, on intraocular pressure and aqueous dynamics in humans. AB - AIMS: To study the effects of topical nipradilol, a non-selective beta blocker with alpha blocking and nitroglycerin-like activities, on intraocular pressure (IOP) and aqueous humour dynamics in normal humans and in patients with ocular hypertension. METHODS: Nipradilol (0.06%, 0.125%, 0.25%, 0.5%) was applied to normal volunteers (n = 12) to test for IOP lowering effects. In a second group of normal volunteers (n = 11), nipradilol (0.125% and 0.25%) and timolol (0. 5%) were compared for IOP lowering effects. After a single administration of 0.25% nipradilol, IOP, flare intensity in the anterior chamber, aqueous flow, uveoscleral outflow, tonographic outflow facility, and episcleral venous pressure were either directly measured or mathematically calculated. Topical nipradilol (0.25%) was administered to 24 patients with ocular hypertension twice daily for 8 weeks. RESULTS: Administration of 0.25% nipradilol decreased IOP with a maximum reduction of 4.2 mm Hg lasting 12 hours. A single instillation of both 0.25% nipradilol and 0.5% timolol reduced the IOP in normotensive human subjects to the same degree. A single instillation of 0.25% nipradilol decreased the aqueous flow rate in the treated eye by 20%. Nipradilol produced no significant effect in tonographic outflow facility or episcleral venous pressure, but uveoscleral outflow was increased. In patients with ocular hypertension, twice daily instillation of 0.25% nipradilol decreased IOP without tachyphylaxis for the 8 week test period. CONCLUSION: Topical nipradilol (0.25%) reduced IOP by decreasing the aqueous flow rate and probably also by increasing uveoscleral outflow. Nipradilol should be further investigated as a new antiglaucoma drug. PMID- 10684842 TI - Intraocular penetration of vancomycin eye drops after application to the medial canthus with closed lids. AB - AIMS: To investigate the intraocular penetration of vancomycin eye drops and to compare the conventional method of drop instillation to the lower cul de sac with applying drops to the medial canthus with closed lids. METHODS: This prospective randomised trial evaluated 53 eyes of 53 patients who had undergone extracapsular cataract extraction (ECCE) with intraocular lens implantation. Vancomycin (50 mg/ml) eye drops were applied to either the lower cul de sac with open lids (conventional method), or to the medial canthus with the patient in a supine position and with closed lids. After paracentesis performed during ECCE, an aqueous humour sample was taken and vancomycin concentration was measured using the TDX vancomycin assay (fluorescence polarisation immunoassay). RESULTS: Vancomycin concentration in the anterior chamber were above the minimal inhibitory concentration for Gram positive bacteria in the two methods of drop instillation examined (2.04 (SD 1.9) microg/ml and 1.49 (1.1) microg/ml in the open and closed methods, respectively (p =0.202)). CONCLUSIONS: Vancomycin (50 mg/ml) reaches therapeutic concentration in the anterior chamber after topical drop application. Comparable concentrations were reached when drops were applied in either the lower cul de sac or to the medial canthus with closed lids. The latter method is proposed as likely to improve patient compliance. PMID- 10684843 TI - Retinal ganglion cell death in experimental glaucoma. AB - AIMS: To determine whether parasol retinal ganglion cells (magnocellular pathway) are selectively lost in the primate model of glaucoma. METHODS: Ocular hypertension was induced in one eye of six Macaca fascicularis monkeys for 6-14 weeks. The retinal ganglion cells in these eyes were labelled retrogradely with the tracer horseradish peroxidase (HRP) implanted into the optic nerve and subsequently examined in retinal whole mount preparations. The degree of retinal ganglion cell loss was estimated from Nissl stained tissue by comparison with the contralateral untreated control eye. RESULTS: In the three glaucomatous retinas with the best labelling 1282 cells could be classified, of which 182 were parasol cells and 1100 were midget cells. Linear regression analysis did not demonstrate a significant reduction in the proportion of parasol to midget cells with increasing cell loss (regression slope 0.023, 95% CI -0.7 to 0.11). Compared with the control eye the cell soma of the remaining retinal ganglion cells in glaucomatous eyes were reduced in size by 20% for parasol cells (p=0.003) and by 16% for midget cells (p <0.001). CONCLUSION: The results of this study do not support the hypothesis that selective loss of parasol retinal ganglion cells occurs in experimental glaucoma. In addition, the change in cell soma size distributions following ocular hypertension suggests that both parasol and midget retinal ganglion cells undergo shrinkage before cell death. PMID- 10684844 TI - Age related changes in the non-collagenous components of the extracellular matrix of the human lamina cribrosa. AB - AIMS: To investigate age related alterations in the non-collagenous components of the human lamina cribrosa. METHODS: Fibronectin, elastin, and glial fibrillary acidic protein (GFAP) staining were assessed in young and old laminae cribrosae. An age range (7 days to 96 years) of human laminae cribrosae were analysed for lipid content (n=9), cellularity (n=28), total sulphated glycosaminoglycans (n=28), elastin content (n=9), and water content (n=56), using chloroform methanol extraction, fluorimetry, the dimethylmethylene blue assay, and ion exchange chromatography, respectively. RESULTS: Qualitatively, an increase in elastin and a decrease in fibronectin and GFAP were demonstrated when young tissue was compared with the elderly. Biochemical analysis of the ageing human lamina cribrosa demonstrated that elastin content increased from 8% to 28% dry tissue weight, total sulphated glycosaminoglycans decreased, and lipid content decreased from 45% to 25%. There were no significant changes in total cellularity or water content. CONCLUSION: These alterations in composition may be indicative of the metabolic state of the lamina cribrosa as it ages, and may contribute to changes in mechanical integrity. Such changes may be implicated in the susceptibility of the elderly lamina cribrosa and also its response to glaucomatous optic neuropathy. PMID- 10684845 TI - Age related compliance of the lamina cribrosa in human eyes. AB - AIMS: To investigate changes in the mechanical compliance of ex vivo human lamina cribrosa with age. METHODS: A laser scanning confocal microscope was used to image the surface of the fluorescently labelled lamina cribrosa in cadaver eyes. A method was developed to determine changes in the volume and strain of the lamina cribrosa created by increases in pressure. The ability of the lamina cribrosa to reverse its deformation on removal of pressure was also measured. RESULTS: Volume and strain measurements both demonstrated that the lamina cribrosa increased in stiffness with age and the level of pressure applied. The ability of the lamina cribrosa to regain its original shape and size on removal of pressure appeared to decrease with age, demonstrating an age related decrease in resilience of the lamina cribrosa. CONCLUSIONS: The mechanical compliance of the human lamina cribrosa decreased with age. Misalignment of compliant cribriform plates in a young eye may exert a lesser stress on nerve axons, than that exerted by the rigid plates of an elderly lamina cribrosa. The resilience of the lamina cribrosa also decreased with age, suggesting an increased susceptibility to plastic flow and permanent deformation. Such changes may be of importance in the explanation of age related optic neuropathy in primary open angle glaucoma. PMID- 10684846 TI - Effect of spectacles on changes of spherical hypermetropia in infants who did, and did not, have strabismus. AB - AIM: To explore why emmetropisation fails in children who have strabismus. METHODS: 289 hypermetropic infants were randomly allocated spectacles and followed. Changes in spherical hypermetropia were compared in those who had strabismus and those who did not. The effect of wearing glasses on these changes was assessed using t tests and regression analysis. RESULTS: Mean spherical hypermetropia decreased in both eyes of "normal" children (p<0.001). The consistent wearing of glasses impeded this process in both eyes (p<0.007). In the children with strabismus, there were no significant changes in either eye, irrespective of treatment (p>0. 05). CONCLUSIONS: In contrast with normal infants, neither eye of those who had strabismus emmetropised, irrespective of whether the incoming vision was clear or blurred. It is suggested that these eyes did not "recognise" the signal of blurred vision, and that they remained long sighted because they were destined to squint. Hence, the children did not squint because they were long sighted, and glasses did not prevent them squinting. PMID- 10684847 TI - Extended wear contact lens related bacterial keratitis. AB - AIMS: To report the clinical findings and visual outcome of patients with extended wear contact lens (EWCL) related bacterial keratitis. METHODS: 11 cases with EWCL related bacterial keratitis were included. Corneal scrapings were obtained for cytology and cultures. RESULTS: Nine patients had unilateral bacterial keratitis and two patients showed bilateral involvement. Corneal scrapings revealed Pseudomonas aeruginosa in seven patients, Staphylococcus aureus coagulase positive in one patient, and Staphylococcus epidermidis in three patients. CONCLUSION: EWCLs may be associated with bacterial keratitis and may result in visual loss. Dispensing contact lenses by optometrists should be performed in consultation with ophthalmologists. PMID- 10684848 TI - Lacrimal drainage surgery in Wegener's granulomatosis. AB - AIM: To examine the results of open lacrimal surgery in patients with Wegener's granulomatosis. METHODS: A retrospective review of patients with Wegener's granulomatosis who underwent lacrimal surgery over a 17 year period. RESULTS: 11 patients were identified and a total of 14 primary dacryocystorhinostomies (DCR) and one revisional DCR were performed; symptomatic relief was achieved in 13/14 operations and one patient required revisional surgery for persistent symptoms. There were no intraoperative and few postoperative complications. CONCLUSIONS: In contrast with some previous reports, open DCR appears to be a safe procedure and it is recommended as a treatment for lacrimal obstruction in patients with Wegener's granulomatosis, but an increase of perioperative immunosuppression is recommended in certain cases. PMID- 10684849 TI - Role of cytokines in the pathogenesis of posterior capsule opacification. PMID- 10684850 TI - Tyrosine-phosphorylated bacterial proteins: Trojan horses for the host cell. PMID- 10684851 TI - Helicobacter pylori CagA protein can be tyrosine phosphorylated in gastric epithelial cells. AB - Attachment of Helicobacter pylori to gastric epithelial cells induces various cellular responses, including the tyrosine phosphorylation of an unknown 145-kD protein and interleukin 8 production. Here we show that this 145-kD protein is the cagA product of H. pylori, an immunodominant, cytotoxin-associated antigen. Epithelial cells infected with various H. pylori clinical isolates resulted in generation of tyrosine-phosphorylated proteins ranging from 130 to 145 kD in size that were also induced in vitro by mixing host cell lysate with bacterial lysate. When epithelial cells were infected with [(35)S]methionine-labeled H. pylori, a radioactive 145-kD protein was detected in the immunoprecipitates with antiphosphotyrosine antibody or anti-CagA (cytotoxin-associated gene A) antibody. Consistently, the 145-kD protein recognized by the anti-CagA and antiphosphotyrosine antibodies was induced in epithelial cells after infection of wild-type H. pylori but not the cagA::Km mutant. Furthermore, the amino acid sequence of the phosphorylated 145-kD protein induced by H. pylori infection was identical to the H. pylori CagA sequence. These results reveal that the tyrosine phosphorylated 145-kD protein is H. pylori CagA protein, which may be delivered from attached bacteria into the host cytoplasm. The identification of the tyrosine-phosphorylated protein will thus provide further insights into understanding the precise roles of CagA protein in H. pylori pathogenesis. PMID- 10684852 TI - The intestinal T cell response to alpha-gliadin in adult celiac disease is focused on a single deamidated glutamine targeted by tissue transglutaminase. AB - The great majority of patients that are intolerant of wheat gluten protein due to celiac disease (CD) are human histocompatibility leukocyte antigen (HLA)-DQ2(+), and the remaining few normally express HLA-DQ8. These two class II molecules are chiefly responsible for the presentation of gluten peptides to the gluten specific T cells that are found only in the gut of CD patients but not of controls. Interestingly, tissue transglutaminase (tTG)-mediated deamidation of gliadin plays an important role in recognition of this food antigen by intestinal T cells. Here we have used recombinant antigens to demonstrate that the intestinal T cell response to alpha-gliadin in adult CD is focused on two immunodominant, DQ2-restricted peptides that overlap by a seven-residue fragment of gliadin. We show that tTG converts a glutamine residue within this fragment into glutamic acid and that this process is critical for T cell recognition. Gluten-specific T cell lines from 16 different adult patients all responded to one or both of these deamidated peptides, indicating that these epitopes are highly relevant to disease pathology. Binding studies showed that the deamidated peptides displayed an increased affinity for DQ2, a molecule known to preferentially bind peptides containing negatively charged residues. Interestingly, the modified glutamine is accommodated in different pockets of DQ2 for the different epitopes. These results suggest modifications of anchor residues that lead to an improved affinity for major histocompatibility complex (MHC), and altered conformation of the peptide-MHC complex may be a critical factor leading to T cell responses to gliadin and the oral intolerance of gluten found in CD. PMID- 10684853 TI - Salmonella-induced apoptosis of infected macrophages results in presentation of a bacteria-encoded antigen after uptake by bystander dendritic cells. AB - Salmonella typhimurium is a gram-negative bacterium that survives and replicates inside vacuolar compartments of macrophages. Infection of macrophages with S. typhimurium grown under conditions allowing expression of the type III secretion system results in apoptotic death of the infected cells. Here, we show that infection of bone marrow-derived macrophages (MPhi) with wild-type S. typhimurium 14028 results in presentation of epitopes derived from a bacteria-encoded antigen on major histocompatibility complex (MHC) class I and MHC class II molecules after internalization of apoptotic MPhi by bystander dendritic cells (DCs). In contrast, infection of MPhi with the phoP constitutive mutant strain CS022, which does not induce apoptosis in infected MPhi, does not result in presentation of a bacteria-derived antigen by bystander DCs unless the infected MPhi are induced to undergo apoptosis by treatment with lipopolysaccharide and ATP. DCs appear to be unique in their ability to present antigens derived from MPhi induced to undergo apoptosis by Salmonella, as bystander MPhi are not capable of presenting the bacteria-derived antigen despite the fact that they efficiently internalize the apoptotic cells. These data suggest that apoptosis induction by bacterial infection of MPhi may not be a quiescent death that allows the bacteria to escape recognition by the immune system, but rather may contribute to an antimicrobial immune response upon engulfment by bystander DCs. PMID- 10684854 TI - Identification of NY-ESO-1 epitopes presented by human histocompatibility antigen (HLA)-DRB4*0101-0103 and recognized by CD4(+) T lymphocytes of patients with NY ESO-1-expressing melanoma. AB - NY-ESO-1 is a member of the cancer-testis family of tumor antigens that elicits strong humoral and cellular immune responses in patients with NY-ESO-1-expressing cancers. Since CD4(+) T lymphocytes play a critical role in generating antigen specific cytotoxic T lymphocyte and antibody responses, we searched for NY-ESO-1 epitopes presented by histocompatibility leukocyte antigen (HLA) class II molecules. Autologous monocyte-derived dendritic cells of cancer patients were incubated with recombinant NY-ESO-1 protein and used in enzyme-linked immunospot (ELISPOT) assays to detect NY-ESO-1-specific CD4(+) T lymphocyte responses. To identify possible epitopes presented by distinct HLA class II alleles, overlapping 18-mer peptides derived from NY-ESO-1 were synthetized and tested for recognition by CD4(+) T lymphocytes in autologous settings. We identified three NY-ESO-1-derived peptides presented by DRB4*0101-0103 and recognized by CD4(+) T lymphocytes of two melanoma patients sharing these HLA class II alleles. Specificity of recognition was confirmed by proliferation assays. The characterization of HLA class II-restricted epitopes will be useful for the assessment of spontaneous and vaccine-induced immune responses of cancer patients against defined tumor antigens. Further, the therapeutic efficacy of active immunization using antigenic HLA class I-restricted peptides may be improved by adding HLA class II-presented epitopes. PMID- 10684855 TI - Promyelocytic leukemia protein (PML) and Daxx participate in a novel nuclear pathway for apoptosis. AB - The promyelocytic leukemia protein (PML) gene of acute promyelocytic leukemia (APL) encodes a cell growth and tumor suppressor essential for multiple apoptotic signals. Daxx was identified as a molecule important for the cytoplasmic transduction of the Fas proapoptotic stimulus. Here, we show that upon mitogenic activation of mature splenic lymphocytes, Daxx is dramatically upregulated and accumulates in the PML nuclear body (NB) where PML and Daxx physically interact. In the absence of PML, Daxx acquires a dispersed nuclear pattern, and activation induced cell death of splenocytes is profoundly impaired. PML inactivation results in the complete abrogation of the Daxx proapoptotic ability. In APL cells, Daxx is delocalized from the NB. Upon retinoic acid treatment, which induces disease remission in APL, Daxx relocalizes to the PML NBs. These results indicate that PML and Daxx cooperate in a novel NB-dependent pathway for apoptosis and shed new light in the role of PML in tumor suppression. PMID- 10684856 TI - Cbl suppresses B cell receptor-mediated phospholipase C (PLC)-gamma2 activation by regulating B cell linker protein-PLC-gamma2 binding. AB - Accumulating evidence indicates that the Cbl protein plays a negative role in immune receptor signaling; however, the mode of Cbl action in B cell receptor (BCR) signaling still remains unclear. DT40 B cells deficient in Cbl showed enhanced BCR-mediated phospholipase C (PLC)-gamma2 activation, thereby leading to increased apoptosis. A possible explanation for the involvement of Cbl in PLC gamma2 activation was provided by findings that Cbl interacts via its Src homology 2 (SH2) domain with B cell linker protein (BLNK) after BCR ligation. BLNK is a critical adaptor molecule for PLC-gamma2 tyrosine phosphorylation through its binding to the PLC-gamma2 SH2 domains. As a consequence of the interaction between Cbl and BLNK, the BCR-induced recruitment of PLC-gamma2 to BLNK and the subsequent PLC-gamma2 tyrosine phosphorylation were inhibited. Thus, our data suggest that Cbl negatively regulates the PLC-gamma2 pathway by inhibiting the association of PLC-gamma2 with BLNK. PMID- 10684857 TI - CD40 ligand (CD154) triggers a short-term CD4(+) T cell activation response that results in secretion of immunomodulatory cytokines and apoptosis. AB - Signals generated through CD28-B7 and CD40 ligand (CD40L)-CD40 interactions have been shown to be crucial for the induction of long-term allograft survivability. We have recently demonstrated that humanized anti-CD40L (hu5C8) prevents rejection of mismatched renal allografts in primates. To investigate potential mechanisms of CD40L-induced allograft acceptance, we coimmobilized hu5C8 with suboptimal amounts of anti-CD3 to stimulate CD4(+) T cells. We now report that anti-CD3/CD40L costimulation results in CD28-independent activation and subsequent deletion of resting T cells. Coligation of CD3 and CD40L increased expression of CD69, CD25, and CD54 on CD4(+) T cells. We also found that costimulation with anti-CD3/CD40L resulted in enhanced production of interleukin (IL)-10, interferon gamma, and tumor necrosis factor alpha but not IL-2 or IL-6. Interestingly, after several days, anti-CD3/CD40L-mediated activation was followed by apoptosis in a significant population of cells. Consistent with that observation, anti-CD3/CD40L did not enhance the antiapoptotic proteins Bcl-2 and Bcl-xL. Further, the addition of CD28 at 24 h failed to rescue those cells induced to die after costimulation with anti-CD3/CD40L. Together, these data suggest that the graft-sparing effect of hu5C8 in vivo may result in part from early and direct effects on CD4(+) T cells, including a vigorous induction of immunomodulatory cytokines and/or apoptosis of allograft-specific T cells. PMID- 10684858 TI - Differential tumor surveillance by natural killer (NK) and NKT cells. AB - Natural tumor surveillance capabilities of the host were investigated in six different mouse tumor models where endogenous interleukin (IL)-12 does or does not dictate the efficiency of the innate immune response. Gene-targeted and lymphocyte subset-depleted mice were used to establish the relative importance of natural killer (NK) and NK1.1(+) T (NKT) cells in protection from tumor initiation and metastasis. In the models examined, CD3(-) NK cells were responsible for tumor rejection and protection from metastasis in models where control of major histocompatibility complex class I-deficient tumors was independent of IL-12. A protective role for NKT cells was only observed when tumor rejection required endogenous IL-12 activity. In particular, T cell receptor Jalpha281 gene-targeted mice confirmed a critical function for NKT cells in protection from spontaneous tumors initiated by the chemical carcinogen, methylcholanthrene. This is the first description of an antitumor function for NKT cells in the absence of exogenously administered potent stimulators such as IL-12 or alpha-galactosylceramide. PMID- 10684859 TI - Fcgamma receptor-mediated phagocytosis in macrophages lacking the Src family tyrosine kinases Hck, Fgr, and Lyn. AB - Macrophage Fcgamma receptors (FcgammaRs) mediate the uptake and destruction of antibody-coated viruses, bacteria, and parasites. We examined FcgammaR signaling and phagocytic function in bone marrow-derived macrophages from mutant mice lacking the major Src family kinases expressed in these cells, Hck, Fgr, and Lyn. Many FcgammaR-induced functional responses and signaling events were diminished or delayed in these macrophages, including immunoglobulin (Ig)G-coated erythrocyte phagocytosis, respiratory burst, actin cup formation, and activation of Syk, phosphatidylinositol 3-kinase, and extracellular signal-regulated kinases 1 and 2. Significant reduction of IgG-dependent phagocytosis was not seen in hck( )(/)-fgr(-)(/)- or lyn(-)(/)- cells, although the single mutant lyn(-)(/)- macrophages did manifest signaling defects. Thus, Src family kinases clearly have roles in two events leading to FcgammaR-mediated phagocytosis, one involving initiation of actin polymerization and the second involving activation of Syk and subsequent internalization. Since FcgammaR-mediated phagocytosis did occur at modest levels in a delayed fashion in triple mutant macrophages, these Src family kinases are not absolutely required for uptake of IgG-opsonized particles. PMID- 10684860 TI - Interleukin 4-producing CD4 T cells arise from different precursors depending on the conditions of antigen exposure in vivo. AB - The precursor origin of T helper (Th) cell subsets in vivo has been difficult to study and remains poorly investigated. We have previously shown that chronic administration of soluble protein antigen induces selective development of antigen-specific CD4 Th2 cells in genetically predisposed mouse strains. To analyze the origin of effector T cells in this model, we designed a competitive polymerase chain reaction-based approach to track public BV-J rearrangement expressed by CD4 T cells specific for hen egg white lysozyme (HEL) in BALB/c mice. We show that public T cell clones are predominantly associated with type 1 or 2 effector Th cells recovered after primary immunization in complete or incomplete Freund's adjuvant, respectively. Conversely, continuous administration of soluble antigen, which induces strong memory Th2 response, is associated with a dose-dependent reduction of public clone size by a mechanism resembling clonal anergy. Thus, soluble HEL-induced Th2 cells do not express the public complementarity determining region 3 motifs characteristic of immunogenic challenge in the presence of adjuvant. These results demonstrate that there are multiple pathways of induction of Th2 responses depending on the condition of antigen exposure in vivo, i.e., clonal immune deviation versus recruitment of a different pool of precursor cells. PMID- 10684861 TI - T cell receptor complementarity determining region 3 length analysis reveals the absence of a characteristic public T cell repertoire in neonatal tolerance. The response in the "tolerant" mouse within the residual repertoire is quantitatively similar but qualitatively different. AB - All adult BALB/c mice immunized with hen egg white lysozyme (HEL) or its dominant determinant, peptide (p)106-116, mount a T cell response using a "public" Vbeta8.2Jbeta1.5 T cell clone. Neonatal exposure to tolerance-inducing doses of antigen can drastically diminish responsiveness in the draining lymph nodes but not in the spleens of animals challenged as adults with the cognate antigen. To determine the role of T cell deletion or anergy within the mechanisms of observed neonatal "tolerance," we treated neonatal BALB/c mice with HEL and directly followed the characteristic public clone using complementarity determining region 3 length T cell repertoire analysis. Our results confirm that despite intraperitoneal injection of neonates with a high dose of HEL emulsified in incomplete Freund's adjuvant, a strong splenic proliferative response to HEL was observed upon recall. However, the adult splenic T cell response of these neonatally treated mice lacked the usual Vbeta8.2Jbeta1.5 public clone characteristic of HEL-primed BALB/c mice. After challenge with HEL-complete Freund's adjuvant as adults, immunoglobulin (Ig)G2a isotype antibody was drastically reduced, and IgG1 was found to be the predominant anti-HEL IgG isotype expressed, indicating a deviation of cytokine response toward T helper type 2. 5-wk-old mice, nasally instilled with tolerogenic doses of HEL p106-116, also showed significant inhibition of this public T cell expansion. These results demonstrate that during neonatal and adult nasal tolerance induction, deletion/anergy removes the public clone, exposing a response of similar specificity but that is characterized by the T helper type 2 phenotype and a splenic residence. PMID- 10684862 TI - Lck domains differentially contribute to pre-T cell receptor (TCR)- and TCR alpha/beta-regulated developmental transitions. AB - Maturational changes at the CD4(-)CD8(-) double negative (DN) to CD4(+)CD8(+) double positive (DP) transition are dependent on signals generated via the pre-T cell receptor (TCR) and the nonreceptor protein tyrosine kinase p56(lck) (Lck). How Lck activities are stimulated or relayed after pre-TCR formation remains obscure. Our structure-function mapping of Lck thymopoietic properties reveals that the noncatalytic domains of Lck are specialized to signal efficient cellular expansion at DN to DP transition. Moreover, although substitution of the Lck catalytic domain with FynT sequences minimally impacts DP development, single positive thymocytes are most efficiently produced in the presence of kinases containing both the NH(2)-terminal and catalytic regions of Lck. These findings demonstrate that the Lck structure is uniquely adapted to mediate signals at both major transitions in thymopoiesis. PMID- 10684863 TI - Induction and suppression of an autoimmune disease by oligomerized T cell epitopes: enhanced in vivo potency of encephalitogenic peptides. AB - T cell epitope peptides derived from proteolipid protein (PLP139-151) or myelin basic protein (MBP86-100) induce experimental autoimmune encephalomyelitis (EAE) in "susceptible" strains of mice (e.g., SJL/J). In this study, we show that the encephalitogenic effect of these epitopes when injected subcutaneously in complete Freund's adjuvant was significantly enhanced if administered to the animal in a multimerized form as a T cell epitope oligomer (i.e., as multiple repeats of the peptide epitope, such as 16-mers). Oligomer-treated SJL/J mice developed EAE faster and showed a more severe progression of the disease than animals treated with peptide alone. In addition, haplotype-matched B10.S mice, "resistant" to EAE induction by peptide, on injection of 16-mers developed a severe form of EAE. Even more striking, however, was the dramatic suppression of incidence and severity of the disease, seen after single intravenous injections of only 50 microg of the PLP139-151 16-mer, administered to SJL/J mice 7 d after the induction of the disease. Although relapse occurred at about day 45, an additional injection several days before that maintained the suppression. Importantly, the specific suppressive effect of oligomer treatment was also evident if EAE was induced with spinal cord homogenate instead of the single peptide antigen. By contrast, the PLP139-151 peptide accelerated rather than retarded the progression of disease. PMID- 10684864 TI - B1 B lymphocytes play a critical role in host protection against lymphatic filarial parasites. AB - Host defense against multicellular, extracellular pathogens such as nematode parasites is believed to be mediated largely, if not exclusively, by T lymphocytes. During our investigations into the course of Brugia malayi and Brugia pahangi infections in immunodeficient mouse models, we found that mice lacking B lymphocytes were permissive for Brugian infections, whereas immunocompetent mice were uniformly resistant. Mice bearing the Btk(xid) mutation were as permissive as those lacking all B cells, suggesting that the B1 subset may be responsible for host protection. Reconstitution of immunodeficient recombination activating gene (Rag)-1(-/)- mice with B1 B cells conferred resistance, even in the absence of conventional B2 lymphocytes and most T cells. These results suggest that B1 B cells are necessary to mediate host resistance to Brugian infection. Our data are consistent with a model wherein early resistance to B. malayi is mediated by humoral immune response, with a significant attrition of the incoming infectious larval load. Sterile clearance of the remaining parasite burden appears to require cell-mediated immunity. These data raise the possibility that the identification of molecule(s) recognized by humoral immune mechanisms might help generate prophylactic vaccines. PMID- 10684865 TI - Inhibition of interleukin 7 receptor signaling by antigen receptor assembly. AB - After the productive rearrangement of immunoglobulin (Ig) heavy chain genes, precursor (pre-)B lymphocytes undergo a limited number of cell divisions in response to interleukin (IL)-7. Here, we present evidence that this phase of IL-7 dependent expansion is constrained by an inhibitory signal initiated by antigen receptor assembly. A line of pre-B cells from normal murine bone marrow that expresses a mu heavy chain with a D-proximal V(H)7183.2 region divides continuously in IL-7. IL-7 responsiveness ceases upon differentiation to the mu(1), kappa(1) stage, despite continuing expression of the IL-7 receptor (IL 7R), suggesting that antigen receptor assembly inhibits IL-7 responsiveness. This is confirmed by introduction of a rearranged lambda light chain gene, which inhibits proliferative signaling through the IL-7R. Inhibition is specific to the IL-7R, because it is overcome by replacement of the IL-7R cytoplasmic domain with corresponding sequences from the closely related IL-2Rbeta chain. Alteration of a single tyrosine residue, Tyr410, in the IL-7R cytoplasmic domain to phenylalanine also prevents the inhibition of proliferation after antigen receptor assembly. Thus, the loss of IL-7 responsiveness after antigen receptor assembly may be mediated through the recruitment of an inhibitory molecule to this residue. Our findings identify a novel mechanism that limits cytokine-dependent proliferation during B lymphopoiesis. This mechanism may be essential for the proper regulation of peripheral B lymphocyte numbers. PMID- 10684866 TI - The role of aquaporins in dendritic cell macropinocytosis. AB - Immature dendritic cells (DCs) constitutively take up large volumes of fluid by macropinocytosis and concentrate the macrosolutes in the endocytic compartment. This concentration mechanism that is the basis of their high capacity to present soluble antigens requires that DCs be capable of rapidly exchanging water across their membranes. We report that two members of the aquaporin family, AQP3 and AQP7, are expressed in immature DCs and are downregulated after maturation. Treatment of DCs with p-chloromercuribenzenesulphonate (pCMBS), a mercuric drug that blocks aquaporins, inhibited uptake and concentration of macrosolutes taken up by fluid phase endocytosis and led to dramatic cell swelling. In contrast, pCMBS did not affect receptor-mediated endocytosis via the mannose receptor. These findings indicate that aquaporins represent essential elements of a volume control mechanism that allows DCs to concentrate macrosolutes taken up via macropinocytosis. PMID- 10684867 TI - Neurons regulate extracellular levels of amyloid beta-protein via proteolysis by insulin-degrading enzyme. AB - Progressive cerebral accumulation of amyloid beta-protein (Abeta) is an early and invariant feature of Alzheimer's disease. Little is known about how Abeta, after being secreted, is degraded and cleared from the extracellular space of the brain. Defective Abeta degradation could be a risk factor for the development of Alzheimer's disease in some subjects. We reported previously that microglial cells release substantial amounts of an Abeta-degrading protease that, after purification, is indistinguishable from insulin-degrading enzyme (IDE). Here we searched for and characterized a role for IDE in Abeta degradation by neurons, the principal cell type that produces Abeta. Whole cultures of differentiated pheochromocytoma (PC12) cells and primary rat cortical neurons actively degraded endogenously secreted Abeta via IDE. However, unlike that in microglia, IDE in differentiated neurons was not released but localized to the cell surface, as demonstrated by biotinylation. Undifferentiated PC12 cells released IDE into their medium, whereas after differentiation, IDE was cell associated but still degraded Abeta in the medium. Overexpression of IDE in mammalian cells markedly reduced the steady-state levels of extracellular Abeta(40) and Abeta(42), and the catalytic site mutation (E111Q) abolished this effect. We observed a novel membrane-associated form of IDE that is approximately 5 kDa larger than the known cytosolic form in a variety of cells, including differentiated PC12 cells. Our results support a principal role for membrane-associated and secreted IDE isoforms in the degradation and clearance of naturally secreted Abeta by neurons and microglia. PMID- 10684868 TI - Declines in mRNA expression of different subunits may account for differential effects of aging on agonist and antagonist binding to the NMDA receptor. AB - The purpose of the present study was to determine whether some of the age-related changes that occur in binding to the NMDA receptor complex can be accounted for by changes in subunit expression during the aging process. In situ hybridization for the NMDA subunits zeta1, epsilon1, and epsilon2, and receptor autoradiography, using the agonist glutamate and the competitive antagonist [(+/ )-2-carboxypiperazin-4-yl] propyl-1-phosphonic acid (CPP), were performed on sections from C57Bl/6 mice representing three different age groups (3, 10, and 30 months of age). There was a significant overall decrease between 3 and 30 month olds in the density of mRNA for the zeta1 subunit in the cortex and hippocampus, but only a few individual brain regions exhibited significant declines. The mRNA for the epsilon2 subunit was significantly decreased in a majority of cortical regions and in the dentate granule cells. Emulsion analysis indicated that the change in the density of epsilon2 subunit mRNA in the inner frontal cortex was primarily attributable to a decrease in the amount of messages per cell. Age related changes in mRNA density of the epsilon2 subunit correlated with changes in NMDA-displaceable [(3)H]glutamate binding, and mRNA density changes in the zeta1 subunit showed a significant relationship with changes in [(3)H]CPP binding in the 30-month-old mice. These results suggest that changes during aging in the expression of different subunits of the NMDA receptor may account for the differential effects of aging on agonist versus antagonist binding to the NMDA binding site. PMID- 10684869 TI - beta subunits modulate alternatively spliced, large conductance, calcium activated potassium channels of avian hair cells. AB - Electrical tuning confers frequency selectivity onto sensory hair cells in the auditory periphery of frogs, turtles, and chicks. The resonant frequency is determined in large part by the number and kinetics of large conductance, calcium activated potassium (BK) channels. BK channels in hair cells are encoded by the alternatively spliced slo gene and may include an accessory beta subunit. Here we examine the origins of kinetic variability among BK channels by heterologous expression of avian cochlear slo cDNAs. Four alternatively spliced forms of the slo-alpha gene from chick hair cells were co-expressed with accessory beta subunits (from quail cochlea) by transient transfection of human embryonic kidney 293 cells. Addition of the beta subunit increased steady-state calcium affinity, raised the Hill coefficient for calcium binding, and slowed channel deactivation rates, resulting in eight functionally distinct channels. For example, a naturally occurring splice variant containing three additional exons deactivated 20-fold more slowly when combined with beta. Deactivation kinetics were used to predict tuning frequencies and thus tonotopic location if hair cells were endowed with each of the expressed channels. All beta-containing channels were predicted to lie within the apical (low-frequency) 30% of the epithelium, consistent with previous in situ hybridization studies. Individual slo-alpha exons would be found anywhere within the apical 70%, depending on the presence of beta, and other alternative exons. Alternative splicing of the slo-alpha channel message provides intrinsic variability in gating kinetics that is expanded to a wider range of tuning by modulation with beta subunits. PMID- 10684870 TI - Coexpression of cloned alpha(1B), beta(2a), and alpha(2)/delta subunits produces non-inactivating calcium currents similar to those found in bovine chromaffin cells. AB - Chromaffin cells express N-type calcium channels identified on the basis of their high sensitivity to block by omega-conotoxin GVIA (omega-CgTx GVIA). In contrast to neuronal N-type calcium currents that inactivate during long depolarizations and that require negative holding potentials to remove inactivation, many chromaffin cells exhibit N-type calcium channel currents that show little inactivation during maintained depolarizations and that exhibit no decrease in channel availability at depolarized holding potentials. N-type calcium channels are thought to be produced by combination of the pore-forming alpha(1B) subunit and accessory beta and alpha(2)/delta subunits. To examine the molecular composition of the non-inactivating N-type calcium channel, we cloned the alpha(1B) and accessory beta (beta(1b), beta(1c,) beta(2a), beta(2b), and beta(3a)) subunits found in bovine chromaffin cells. Expression of the subunits in either Xenopus oocytes or human embryonic kidney 293 cells produced high threshold calcium currents that were blocked by omega-CgTx GVIA. Coexpression of bovine alpha(1B) with beta(1b), beta(1c), beta(2b), or beta(3a) produced currents that were holding potential dependent. In contrast, coexpression of bovine alpha(1B) with beta(2a) produced holding potential-independent calcium currents that closely mimicked native non-inactivating currents, suggesting that non inactivating N-type channels consist of bovine alpha(1B), alpha(2)/delta, and beta(2a). PMID- 10684871 TI - Estrogen-induced activation of the mitogen-activated protein kinase cascade in the cerebral cortex of estrogen receptor-alpha knock-out mice. AB - We have shown previously in the developing cerebral cortex that estrogen elicits the rapid and sustained activation of multiple signaling proteins within the mitogen-activated protein (MAP) kinase cascade, including B-Raf and extracellular signal-regulated kinase (ERK). Using estrogen receptor (ER)-alpha gene-disrupted (ERKO) mice, we addressed the role of ER-alpha in mediating this action of estrogen in the brain. 17beta-Estradiol increased B-Raf activity and MEK (MAP kinase/ERK kinase)-dependent ERK phosphorylation in cerebral cortical explants derived from both ERKO and their wild-type littermates. The ERK response was stronger in ERKO-derived cultures but, unlike that of wild-type cultures, was not blocked by the estrogen receptor antagonist ICI 182,780. Surprisingly, both the ER-alpha selective ligand 16alpha-iodo-17beta-estradiol and the ER-beta selective ligand genistein failed to elicit ERK phosphorylation, suggesting that a different mechanism or receptor may mediate estrogen-induced ERK phosphorylation in the cerebral cortex. Interestingly, the transcriptionally inactive stereoisomer 17alpha-estradiol did elicit a strong induction of ERK phosphorylation, which, together with the inability of the ER-alpha- and ER-beta selective ligands to elicit ERK phosphorylation, and of ICI 182,780 to block the actions of estradiol in ERKO cultures, supports the hypothesis that a novel, estradiol-sensitive and ICI-insensitive estrogen receptor may mediate 17beta estradiol-induced activation of ERK in the brain. PMID- 10684872 TI - Calcium transients in the rhabdomeres of dark- and light-adapted fly photoreceptor cells. AB - The light response of fly photoreceptor cells is modulated by changes in free Ca(2+) concentration. Fly phototransduction and most processes regulating it take place in or very close to the rhabdomere. We therefore measured the kinetics and the absolute values of the free Ca(2+) concentration in the rhabdomere of fly photoreceptor cells in vivo by making use of the natural optics of the fly's eye. We show that Ca(2+) flowing into the rhabdomere after light stimulation of dark adapted cells causes fast Ca(2+) transients that reach peak values higher than 200 microM in <20 msec. Approximately 500 msec later, the free Ca(2+) concentration has declined again to approximately 20 microM. The duration of the Ca(2+) transients becomes still shorter, and their size reduced, when the photoreceptor cell is light-adapted. This reduction in duration and size of the Ca(2+) transients is graded with the intensity of the adapting light. The kinetics and absolute values of the free calcium concentration found to occur in the rhabdomere are suitable to mediate the fast feedback signals known to act on the fly phototransduction cascade. PMID- 10684873 TI - Reconstitution of muscarinic modulation of the KCNQ2/KCNQ3 K(+) channels that underlie the neuronal M current. AB - Channels from KCNQ2 and KCNQ3 genes have been suggested to underlie the neuronal M-type K(+) current. The M current is modulated by muscarinic agonists via G proteins and an unidentified diffusible cytoplasmic messenger. Using whole-cell clamp, we studied tsA-201 cells in which cloned KCNQ2/KCNQ3 channels were coexpressed with M(1) muscarinic receptors. Heteromeric KCNQ2/KCNQ3 currents were modulated by the muscarinic agonist oxotremorine-M (oxo-M) in a manner having all of the characteristics of modulation of native M current in sympathetic neurons. Oxo-M also produced obvious intracellular Ca(2+) transients, observed by using indo-1 fluorescence. However, modulation of the current remained strong even when Ca(2+) signals were abolished by the combined use of strong intracellular Ca(2+) buffers, an inhibitor of IP(3) receptors, and thapsigargin to deplete Ca(2+) stores. Muscarinic modulation was not blocked by staurosporine, a broad-spectrum protein kinase inhibitor, arguing against involvement of protein kinases. The modulation was not associated with a shift in the voltage dependence of channel activation. Homomeric KCNQ2 and KCNQ3 channels also expressed well and were modulated individually by oxo-M, suggesting that the motifs for modulation are present on both channel subtypes. Homomeric KCNQ2 and KCNQ3 currents were blocked, respectively, at very low and at high concentrations of tetraethylammonium ion. Finally, when KCNQ2 subunits were overexpressed by intranuclear DNA injection in sympathetic neurons, total M current was fully modulated by the endogenous neuronal muscarinic signaling mechanism. Our data further rule out Ca(2+) as the diffusible messenger. The reconstitution of muscarinic modulation of the M current that uses cloned components should facilitate the elucidation of the muscarinic signaling mechanism. PMID- 10684874 TI - Mechanisms of calcium decay kinetics in hippocampal spines: role of spine calcium pumps and calcium diffusion through the spine neck in biochemical compartmentalization. AB - Dendritic spines receive most excitatory inputs in the CNS and compartmentalize calcium. Although the mechanisms of calcium influx into spines have been explored, it is unknown what determines the calcium decay kinetics in spines. With two-photon microscopy we investigate action potential-induced calcium dynamics in spines from rat CA1 pyramidal neurons in slices. The [Ca(2+)](i) in most spines shows two decay kinetics: an initial fast component, during which [Ca(2+)](i) in spines decays to dendritic levels, followed by a slower decay phase in which the spine follows dendritic kinetics. The correlation between [Ca(2+)](i) in spine and dendrite at the breakpoint of the decay kinetics demonstrates diffusional equilibration between spine and dendrite during the slower component. To explain the faster initial decay, we rule out saturation or kinetic effects of endogenous or exogenous buffers and focus instead on (1) active calcium extrusion and (2) buffered diffusion of calcium from spine to dendrite. The presence of an undershoot in most spines indicates that extrusion mechanisms can be intrinsic to the spine. Supporting the two mechanisms, pharmacological blockade of smooth endoplasmic reticulum calcium (SERCA) pumps and the length of the spine neck affect spine decay kinetics. Using a mathematical model, we find that the contribution of calcium pumps and diffusion varies from spine to spine. We conclude that dendritic spines have calcium pumps and that their density and kinetics, together with the morphology of the spine neck, determine the time during which the spine compartmentalizes calcium. PMID- 10684875 TI - Reciprocal inhibitory connections regulate the spatiotemporal properties of intrathalamic oscillations. AB - Mice with an inactivated GABA(A) receptor beta(3) subunit gene have features of Angelman syndrome, including absence-like seizures. This suggests the occurrence of abnormal hypersynchrony in the thalamocortical system. Within the thalamus, the efficacy of inhibitory synapses between thalamic reticular (RE) neurons is selectively compromised, and thalamic oscillations in vitro are prolonged and lack spatial phase gradients (). Here we used computational models to examine how intra-RE inhibition regulates intrathalamic oscillations. A major effect is an abbreviation of network responses, which is caused by long-lasting intra-RE inhibition that shunts recurrent excitatory input. In addition, differential activation of RE cells desynchronizes network activity. Near the slice center, where many cells are initially activated, there is a resultant high level of intra-RE inhibition. This leads to RE cell burst truncation in the central region and a gradient in the timing of thalamocortical cell activity similar to that observed in vitro. Although RE cell burst durations were shortened by this mechanism, there was very little effect on the times at which RE cells began to burst. The above results depended on widespread stimuli that activated RE cells in regions larger than the diameter of intra-RE connections. By contrast, more focal stimuli could elicit oscillations that lasted several cycles and remained confined to a small region. These results suggest that intra-RE inhibition restricts intrathalamic activity to particular spatiotemporal patterns to allow focal recurrent activity that may be relevant for normal thalamocortical function while preventing widespread synchronization as occurs in seizures. PMID- 10684876 TI - dCLOCK is present in limiting amounts and likely mediates daily interactions between the dCLOCK-CYC transcription factor and the PER-TIM complex. AB - In Drosophila melanogaster four circadian clock proteins termed PERIOD (PER), TIMELESS (TIM), dCLOCK (dCLK), and CYCLE (CYC/dBMAL1) function in a transcriptional feedback loop that is a core element of the oscillator mechanism. dCLK and CYC are members of the basic helix-loop-helix (bHLH)/PAS (PER-ARNT-SIM) superfamily of transcription factors and are required for high-level expression of per and tim and repression of dClk, whereas PER and TIM inhibit dCLK-CYC mediated transcription and lead to the activation of dClk. To understand further the dynamic regulation within the circadian oscillator mechanism, we biochemically characterized in vivo-produced CYC, determined the interactions of the four clock proteins, and calculated their absolute levels as a function of time. Our results indicate that throughout a daily cycle the majority of the dCLK present in adult heads stably interacts with CYC, indicating that CYC is the primary in vivo partner of dCLK. dCLK-CYC dimers are bound by PER and TIM during the late evening and early morning, suggesting the formation of a tetrameric complex with impaired transcriptional activity. Although dCLK is present in limiting amounts and CYC is by far the most abundant of the four clock proteins that have been examined, PER and TIM appear to interact preferentially with dCLK. Our results suggest that dCLK is the main component regulating the daily abundance of transcriptionally active dCLK-CYC complexes. PMID- 10684877 TI - Activity and calcium-dependent mechanisms maintain reliable interneuron synaptic transmission in a rhythmic neural network. AB - Inputs from glutamatergic excitatory interneurons (EIN) to motor neurons in the lamprey spinal cord locomotor network exhibit activity-dependent depression during spike trains. The mechanism underlying this depression has been examined here, and its relevance to transmitter release during rhythmic activity has been investigated. The depression of EIN inputs was greater after larger initial EPSPs and reduced in low-calcium Ringer's solution, effects that are consistent with depression caused by depletion of releasable transmitter stores. However, the depression was greater at lower stimulation frequencies and could be reversed by increasing the stimulation frequency. In addition, high-calcium Ringer's solution and the slow intracellular calcium chelator EGTA-AM, which both failed to affect the amplitude of low frequency-evoked EPSPs, reduced and increased the depression, respectively. These results are inconsistent with a simple depletion mechanism but suggest that ongoing activity and calcium-dependent mechanisms oppose depletion. The network relevance of this mechanism was examined using physiologically relevant bursts to simulate EIN spiking during rhythmic activity. Although considerably more EPSPs were evoked than during spike trains, burst evoked EPSPs did not depress. However, single EPSPs evoked at the interburst interval depressed, and burst transmission was disrupted by EGTA-AM, again suggesting the involvement of activity and calcium-dependent mechanisms. By responding to the calcium changes evoked by increased interneuron activity, this mechanism can monitor transmitter requirements caused by EIN spiking, allowing reliable transmission across different patterns of network activity. However, not all types of spinal interneurons exhibit reliable burst transmission, suggesting specificity of this mechanism to a subset of neurons. PMID- 10684878 TI - A fundamental role for the nitric oxide-G-kinase signaling pathway in mediating intercellular Ca(2+) waves in glia. AB - In this study, we highlight a role for the nitric oxide-cGMP-dependent protein kinase (NO-G-kinase) signaling pathway in glial intercellular Ca(2+) wave initiation and propagation. Addition of the NO donor molsidomine (100-500 microM) or puffing aqueous NO onto primary glial cell cultures evoked an increase in [Ca(2+)](i) in individual cells and also local intercellular Ca(2+) waves, which persisted after removal of extracellular Ca(2+). High concentrations of ryanodine (100-200 microM) and antagonists of the NO-G-kinase signaling pathway essentially abrogated the NO-induced increase in [Ca(2+)](i), indicating that NO mobilizes Ca(2+) from a ryanodine receptor-linked store, via the NO-G-kinase signaling pathway. Addition of 10 microM nicardipine to cells resulted in a slowing of the molsidomine-induced rise in [Ca(2+)](i), and inhibition of Mn(2+) quench of cytosolic fura-2 fluorescence mediated by a bolus application of 2 microM aqueous NO to cells, indicating that NO also induces Ca(2+) influx in glia. Mechanical stress of individual glial cells resulted in an increase in intracellular NO in target and neighboring cells and intercellular Ca(2+) waves, which were NO, cGMP, and G-kinase dependent, because incubating cells with nitric oxide synthase, guanylate cyclase, and G-kinase inhibitors, or NO scavengers, reduced Delta[Ca(2+)](i) and the rate of Ca(2+) wave propagation in these cultures. Results from this study suggest that NO-G-kinase signaling is coupled to Ca(2+) mobilization and influx in glial cells and that this pathway plays a fundamental role in the generation and propagation of intercellular Ca(2+) waves in glia. PMID- 10684879 TI - Tachykinin-related peptide and GABA-mediated presynaptic inhibition of crayfish photoreceptors. AB - Off-axis illumination elicits lateral inhibition at the primary visual synapse in crustacea and insects. The evidence suggests that the inhibitory action is presynaptic (i.e., on the photoreceptor terminal) and that the amacrine neurons of the lamina ganglionaris (the first synaptic layer) may be part of the inhibitory pathway. The neurotransmitters and the synaptic mechanisms are unknown. We show by immunocytochemistry that GABA and a tachykinin-related peptide (TRP) are localized in the amacrine neurons of the crayfish lamina ganglionaris. Indirect evidence suggests that GABA and TRP may be colocalized in these neurons. The extensive processes of the amacrine neurons occupy lamina layers containing the terminals of photoreceptors. Application of exogenous GABA and TRP to photoreceptor terminals produces a short-latency, dose-dependent hyperpolarization with a decay time constant on the order of a few seconds. TRP also exhibits actions that evolve over several minutes. These include a reduction of the receptor potential (and the light-elicited current) by approximately 40% and potentiation of the action of GABA by approximately 100%. The mechanisms of TRP action in crayfish are not known, but a plausible pathway is a TRP-dependent elevation of intracellular Ca(2+) that reduces photoreceptor sensitivity in arthropods. Although the mechanisms are not established, the results indicate that in crayfish photoreceptors TRP displays actions on two time scales and can exert profound modulatory control over cell function. PMID- 10684880 TI - NMDA receptor-mediated subthreshold Ca(2+) signals in spines of hippocampal neurons. AB - We have used rapid confocal microscopy to investigate the mechanism of Ca(2+) signals in individual dendritic spines of hippocampal CA1 pyramidal cells. The experiments focused on the signals that occur during single weak synaptic responses that were subthreshold for triggering postsynaptic action potentials. These Ca(2+) signals were not strongly affected by blocking the EPSPs with the AMPA receptor antagonist CNQX. The signals were also not strongly reduced by blocking T-type voltage-gated Ca(2+) channels (VGCCs) with Ni(2+) or by blocking a broad range of VGCCs with intracellular D890. The spine Ca(2+) signals were blocked by NMDA receptor channel (NMDAR) antagonist and had the voltage dependence characteristic of these channels. Neither ryanodine nor cyclopiazonic acid (CPA), substances known to deplete intracellular Ca(2+) stores, substantially reduced the amplitude of synaptically evoked Ca(2+) signals. CPA slowed the recovery phase of Ca(2+) signals in spines produced by synaptic stimulation or by backpropagating action potentials, suggesting a role of intracellular stores in Ca(2+) reuptake. Thus, we find that Ca(2+) release from intracellular stores is not required to produce spine Ca(2+) signals. We conclude that synaptic Ca(2+) signals in spines are primarily caused by Ca(2+) entry through NMDARs. Although these channels are largely blocked by Mg(2+) at voltages near the resting potential, they can nevertheless produce significant Ca(2+) elevation. The resulting Ca(2+) signals are an integral component of individual evoked or spontaneous synaptic events and may be important in the maintenance of synaptic function. PMID- 10684881 TI - Imaging extracellular waves of glutamate during calcium signaling in cultured astrocytes. AB - A growing body of evidence proposes that glial cells have the potential to play a role as modulators of neuronal activity and synaptic transmission by releasing the neurotransmitter glutamate (Arague et al., 1999). We explore the spatial nature of glutamate release from astrocytes with an enzyme-linked assay system and CCD imaging technology. In the presence of glutamate, L-glutamic dehydrogenase (GDH) reduces NAD(+) to NADH, a product that fluoresces when excited with UV light. Theoretically, provided that GDH and NAD(+) are present in the bathing saline, the release of glutamate from stimulated astrocytes can be optically detected by monitoring the accumulation of NADH. Indeed, stimuli that induce a wave of elevated calcium among astrocytes produced a corresponding spread of extracellular NADH fluorescence. Treatment of cultures either with thapsigargin, to deplete internal calcium stores, or with the membrane-permeant calcium chelator BAPTA AM significantly decreased the accumulation of NADH, demonstrating that this fluorometric assay effectively monitors calcium-dependent glutamate release. With a temporal resolution of 500 msec and spatial resolution of approximately 20 micrometer, discrete regions of glutamate release were not reliably resolved. The wave of glutamate release that underlies the NADH fluorescence propagated at an average speed of approximately 26 micrometer/sec, correlating with the rate of calcium wave progression (10-30 micrometer/sec), and caused a localized accumulation of glutamate in the range of 1-100 microM. Further analysis of the fluorescence accumulation clearly demonstrated that glutamate is released in a regenerative manner, with subsequent cells that are involved in the calcium wave releasing additional glutamate. PMID- 10684882 TI - Mechanisms of glutamate metabolic signaling in retinal glial (Muller) cells. AB - Retinal Muller (glial) cells metabolize glucose to lactate, which is preferentially taken up by photoreceptor neurons as fuel for their oxidative metabolism. We explored whether lactate supply to neurons is a glial function controlled by neuronal signals. For this, we used subcellular fluorescence imaging and either amperometric or optical biosensors to monitor metabolic responses simultaneously from mitochondrial and cytosolic compartments of individual Muller cells from salamander retina. Our results demonstrate that lactate production and release is controlled by the combined action of glutamate and NH(4)(+), both at micromolar concentrations. Transport of glutamate by a high affinity carrier can produce in Muller cells a rapid rise of glutamate concentration. In our isolated Muller cells, glutamine synthetase (GS) converted transported glutamate to glutamine that was released. This reaction, predominant when enough NH(4)(+) is available, was limited at micromolar concentrations of NH(4)(+), and more glutamate entered then as substrate into the mitochondrial tricarboxylic acid cycle (TCA). Increased production of glutamine by GS leads to increased utilization of ATP, some of which is generated glycolytically. Methionine sulfoximine, a specific inhibitor of GS, suppressed the stimulatory effect of glutamate and NH(4)(+) on glycolysis and induced massive entry of glutamate into the TCA cycle. The rate of glutamine production also determined the amount of pyruvate transaminated by glutamate to alanine. Lactate, alanine, and glutamine can be taken up and metabolized by photoreceptor neurons. We conclude that a major function of Muller glial cells is to nourish retinal neurons and to metabolize the neurotoxic ammonia and glutamate. PMID- 10684883 TI - Impaired axonal regeneration in alpha7 integrin-deficient mice. AB - The interplay between growing axons and the extracellular substrate is pivotal for directing axonal outgrowth during development and regeneration. Here we show an important role for the neuronal cell adhesion molecule alpha7beta1 integrin during peripheral nerve regeneration. Axotomy led to a strong increase of this integrin on regenerating motor and sensory neurons, but not on the normally nonregenerating CNS neurons. alpha7 and beta1 subunits were present on the axons and their growth cones in the regenerating facial nerve. Transgenic deletion of the alpha7 subunit caused a significant reduction of axonal elongation. The associated delay in the reinnervation of the whiskerpad, a peripheral target of the facial motor neurons, points to an important role for this integrin in the successful execution of axonal regeneration. PMID- 10684884 TI - Surviving granule cells of the sclerotic human hippocampus have reduced Ca(2+) influx because of a loss of calbindin-D(28k) in temporal lobe epilepsy. AB - In mesial temporal lobe epilepsy (mTLE), the predominant form of epilepsy in adults, and in animal models of the disease, there is a conspicuous loss of the intracellular Ca(2+)-binding protein calbindin-D(28k) (CB) from granule cells (GCs) of the dentate gyrus. The role of this protein in nerve cell function is controversial, but here we provide evidence for its role in controlling Ca(2+) influx into human neurons. In patients with Ammon's horn sclerosis (AHS), the loss of CB from GCs markedly increased the Ca(2+)-dependent inactivation of voltage-dependent Ca(2+) currents (I(Ca)), thereby diminishing Ca(2+) influx during repetitive neuronal firing. Introducing purified CB into GCs restored Ca(2+) current inactivation to levels observed in cells with normal CB content harvested from mTLE patients without AHS. Our data are consistent with the possibility of neuroprotection secondary to the CB loss. By limiting Ca(2+) influx through an enhanced Ca(2+)-dependent inactivation of voltage-dependent Ca(2+) channels during prolonged neuronal discharges, the loss of CB may contribute to the resistance of surviving human granule cells in AHS. PMID- 10684885 TI - Serotonin-driven long-range inhibitory connections in the cerebellar cortex. AB - Disturbances of the serotoninergic neuromodulation in the cerebellar cortex have been involved in several types of ataxia, but the physiological action of serotonin in this structure remains poorly understood. We report that in slices of the rat cerebellar vermis, serotonin triggers the firing of an inhibitory interneuron presynaptic to Golgi cells. The Lugaro cell, a neglected interneuronal type, satisfies the expected criteria for this input, whereas basket cells, stellate cells, or Golgi cells do not. Lugaro cells are selectively excited by serotonin, and their firing behavior (sustained steady frequency in the 5-15 Hz range) resembles the pattern of occurrence of serotonin-evoked IPSCs in Golgi cells. Immunohistochemical stainings and single cell reconstructions show that Lugaro cell axons form a parasagittal plexus but also extend long transverse branches that run parallel to the parallel fibers and are partly myelinated. Electrophysiological data suggest that these transverse axons participate in synaptic contacts of the Lugaro cells with Golgi cells, and we calculated that in the intact cerebellum a given Lugaro cell contacts >100 Golgi cells. Serotonin modulation of Lugaro cells may constitute an intracortical switch involved in information patterning at the level of Golgi cells and granule cells populations, and particularly in synchronizations recorded along the transverse axis in vivo. PMID- 10684886 TI - Activation of extracellular signal-regulated protein kinases is associated with a sensitized locomotor response to D(2) dopamine receptor stimulation in unilateral 6-hydroxydopamine-lesioned rats. AB - Evidence indicates that mitogen-activated protein kinase (MAPK) pathways play a crucial role in the neurobiology of the nervous system. In the present study, dopamine receptor-mediated regulation of extracellular signal-regulated kinases (ERKs) was examined in rats in which the nigrostriatal dopaminergic pathway was unilaterally lesioned by 6-hydroxydopamine (6-OHDA). Subcutaneous injections of the D(2) receptor agonist quinpirole significantly increased tyrosine phosphorylated ERK1/2 in lesioned striatum, whereas the D(1) receptor agonist SKF38393 failed to activate ERKs. Quinpirole-induced phosphorylation of ERK1/2 was seen as early as 3 min and peaked at 15 min after the challenge. In parallel, striatal ERK kinase activity, measured by the in vitro kinase assay, was increased 2.5-fold on the lesioned side after the administration of quinpirole. Immunohistochemical examination of brain sections after quinpirole administration revealed significant increases in ERK1/2 immunostaining in perinuclear and intranuclear areas of striatal neurons. This increase was much more pronounced on the lesioned than the intact side. Furthermore, quinpirole-induced contralateral rotation was decreased by 48.7 and 50.7%, respectively, when the striatal ERK pathway was selectively inhibited by a single intrastriatal injection of the MAPK/ERK kinase inhibitor PD098059 or after a continuous 7 d intrastriatal infusion of ERK1/2 antisense oligodeoxynucleotide. The results demonstrate, for the first time, that the ERK signaling pathway is activated in denervated striatum in response to stimulation of D(2) dopamine receptors and that the resulting imbalance in striatal ERK activity contributes, at least in part, to neuronal plasticity that underlies D(2) dopamine receptor-mediated contralateral rotation in unilateral 6-OHDA denervated rats. PMID- 10684887 TI - Embryonic and early fetal development of the human neocortex. AB - Early corticogenesis was studied in human embryos and early fetuses from Carnegie stages 16 to 22 (5-8 gestational weeks) by using immunohistochemistry for Reelin (Reln), calretinin (CR), and glutamic acid decarboxylase (GAD). A first population of Reln-positive cells appears in the neocortical anlage at stage 16 and increases in number at stages 17-18. At stages 19-20, a monolayer of horizontal CR- and GAD-positive, Reln-negative neurons forms in the preplate, whereas Reln-positive cells shift into a subpial position. Another cell class, the pioneer projection neuron, is CR-positive but GAD- and Reln-negative; pioneer cells contribute early corticofugal axons. Pioneer cells first appear below the monolayer at stage 20 and form a pioneer plate at stage 21. The cortical plate (CP) proper emerges at stage 21 and inserts itself within the pioneer plate, which is thus split into a minor superficial component and a larger deep component that presumably corresponds to the subplate. Initial CP neurons are radially organized and mostly CR-negative. Reln-positive cells remain consistently segregated from the pioneer cells and are thus not directly involved in preplate partition. Our data indicate that the neuronal composition of the human neocortical preplate is more complex than generally described and that various neurons participate in a sequence of events that precede the emergence of the CP. PMID- 10684888 TI - Expression of Kv1 potassium channels in mouse hippocampal primary cultures: development and activity-dependent regulation. AB - Excitability and discharge behavior of neurons depends on the highly variable expression pattern of voltage-dependent potassium (Kv) channels throughout the nervous system. To learn more about distribution, development, and activity dependent regulation of Kv channel subunit expression in the rodent hippocampus, we studied the protein expression of members of the Kv1 subfamily in mouse hippocampus in situ and in primary cultures. In adult hippocampus, Kv1 (1-6) channel alpha-subunits were present, whereas at postnatal day 2, none of these proteins could be detected in CA1-CA3 and dentate gyrus. Kv1.1 was the first channel to be observed at postnatal day 6. The delayed postnatal expression and most of the subcellular distribution observed in hippocampal sections were mimicked by cultured hippocampal neurons in which Kv channels appeared only after 10 days in vitro. This developmental upregulation was paralleled by a dramatic increase in total K(+) current, as well as an elevated GABA release in the presence of 4-aminopyridine. Thus, the developmental profile, subcellular localization, and functionality of Kv1 channels in primary culture of hippocampus closely resembles the in situ situation. Impairing secretion by clostridial neurotoxins or blocking activity by tetrodotoxin inhibited the expression of Kv1.1, Kv1.2, and Kv1.4, whereas the other Kv1 channels still appeared. This activity-dependent depression was only observed before the initial appearance of the respective channels and lost after they had been expressed. Our data show that hippocampal neurons in culture are a convenient model to study the developmental expression and regulation of Kv1 channels. The ontogenetic regulation and the activity-dependent expression of Kv1.1, Kv1.2, and Kv1.4 indicate that neuronal activity plays a crucial role for the development of the mature Kv channel pattern in hippocampal neurons. PMID- 10684889 TI - Disruption of the olfactoretinal centrifugal pathway may relate to the visual system defect in night blindness b mutant zebrafish. AB - We describe here a dominant mutation, night blindness b (nbb), which causes an age-related visual system defect in zebrafish. At 4-5 months of age, dark-adapted nbb(+/-) mutants show abnormal visual threshold fluctuations when measured behaviorally. Light sensitizes the animals; thus early dark adaptation of nbb(+/ ) fish is normal. After 2 hr of dark adaptation, however, visual thresholds of nbb(+/-) mutants are raised on average 2-3 log units, and rod system function is not detectable. Electroretinograms recorded from nbb(+/-) mutants are normal, but ganglion cell thresholds are raised in prolonged darkness, suggesting an inner retinal defect. The visual defect of nbb(+/-) mutants may be likely caused by an abnormal olfactoretinal centrifugal innervation; in nbb(+/-) mutants, the olfactoretinal centrifugal projection to the retina is disrupted, and the number of retinal dopaminergic interplexiform cells is reduced. A similar visual defect as shown by nbb(+/-) mutants is observed in zebrafish in which the olfactory epithelium and olfactory bulb have been excised. Homozygous nbb fish display an early onset neural degeneration throughout the CNS and die by 7-8 d of age. PMID- 10684890 TI - Effects of dopamine depletion on visual sensitivity of zebrafish. AB - The visual sensitivity of zebrafish in which the retinal dopaminergic interplexiform cells (DA-IPCs) were destroyed by 6-hydroxydopamine was measured behaviorally. During the first 6-8 min of dark adaptation, visual thresholds of DA-IPC-depleted animals were similar to those of control animals. Thereafter, their visual thresholds were elevated so that by 14-18 min of dark adaptation, they were 2-3 log units above those of control animals. In DA-IPC-depleted animals, the electroretinogram was normal in terms of light sensitivity and waveform, but the light threshold for eliciting a ganglion cell discharge was raised by 1.8 log units as compared with control animals. No obvious rod system function was detected in DA-IPC-depleted animals as measured behaviorally. Partial rescue of the behavioral visual sensitivity loss in DA-IPC-depleted animals occurred when dopamine or a long-acting dopamine agonist (2-amino-6, 7 dihydroxy-1, 2, 3, 4-tetrahydronaphthalene hydrobromide) were injected intraocularly. Our data suggest that the principal visual defect shown by DA-IPC depleted animals is attributable to effects occurring in the inner retina, mainly on rod signals. We also show that dopamine is involved in mediating the effect of the circadian clock on visual sensitivity. PMID- 10684891 TI - Brain-derived neurotrophic factor acutely inhibits AMPA-mediated currents in developing sensory relay neurons. AB - Brain-derived neurotrophic factor (BDNF) is expressed by many primary sensory neurons that no longer require neurotrophins for survival, indicating that BDNF may be used as a signaling molecule by the afferents themselves. Because many primary afferents also express glutamate, we investigated the possibility that BDNF modulates glutamatergic AMPA responses of newborn second-order sensory relay neurons. Perforated-patch, voltage-clamp recordings were made from dissociated neurons of the brainstem nucleus tractus solitarius (nTS), a region that receives massive primary afferent input from BDNF-containing neurons in the nodose and petrosal cranial sensory ganglia. Electrophysiological analysis was combined in some experiments with anterograde labeling of primary afferent terminals to specifically analyze responses of identified second-order neurons. Our data demonstrate that BDNF strongly inhibits AMPA-mediated currents in a large subset of nTS cells. Specifically, AMPA responses were either completely abolished or markedly inhibited by BDNF in 73% of postnatal day (P0) cells and in 82% of identified P5 second-order sensory relay neurons. This effect of BDNF is mimicked by NT-4, but not NGF, and blocked by the Trk tyrosine kinase inhibitor K252a, consistent with a requirement for TrkB receptor activation. Moreover, analysis of TrkB expression in culture revealed a close correlation between the percentage of nTS neurons in which BDNF inhibits AMPA currents and the percentage of neurons that exhibit TrkB immunoreactivity. These data document a previously undefined mechanism of acute modulation of AMPA responses by BDNF and indicate that BDNF may regulate glutamatergic transmission at primary afferent synapses. PMID- 10684892 TI - Afferent regulation of inhibitory synaptic transmission in the developing auditory midbrain. AB - To determine whether afferent innervation regulates the strength of inhibitory connections in the gerbil auditory midbrain, both cochleas were surgically removed in postnatal day 7 animals, before sound-driven activity is first observed. Inhibitory synaptic currents were measured in a brain slice preparation 1-7 d after the ablations. Whole-cell and gramicidin-perforated patch recordings were obtained from inferior colliculus neurons, and IPSCs were evoked by stimulation of the commissure of the inferior colliculus (CIC) or the ipsilateral lateral lemniscus (LL) in the presence of kynurenic acid. Deafferentation led to a 24 mV depolarizing shift in the IPSC equilibrium potential within 1 d of deafferentation. As a consequence, there was a large reduction of IPSC amplitude at a holding potential of -20 mV in neurons from bilaterally ablated animals. Furthermore, both afferent pathways displayed a 50% reduction of the inhibitory synaptic conductance after deafferentation, indicating that driving force was not solely responsible for the decline in IPSC amplitude. When paired pulses were delivered to the LL or CIC pathway in control neurons, the evoked IPSCs exhibited facilitation. However, paired pulse facilitation was nearly eliminated after deafferentation. Thus, normal innervation affects inhibitory synaptic strength by regulating postsynaptic chloride homeostasis and presynaptic transmitter release properties. PMID- 10684893 TI - Cortical cell orientation selectivity fails to develop in the absence of ON center retinal ganglion cell activity. AB - Neuronal activity is necessary for the normal development of visual cortical cell receptive fields. When neuronal activity is blocked, cortical cells fail to develop normal ocular dominance and orientation selectivity. Patterned activity has been shown to play an instructive, rather than merely permissive, role in the segregation of geniculocortical afferents into ocular dominance columns. To test whether normal patterns of activity are necessary to instruct the development of cortical orientation selectivity, we studied ferrets raised without ON-center retinal ganglion cell activity. The ON-center blockade was produced by daily intravitreal injections of DL-2-amino-4-phosphonobutyric acid (APB). Effects of this treatment on the development of orientation selectivity in primary visual cortex were assessed using extracellular electrode recordings and optical imaging. In animals raised with an ON-center blockade starting after visual cortical cells are visually driven but still poorly tuned for orientation, cortical cell responsivity was maintained, but no maturation of orientation selectivity was seen. No recovery of orientation tuning was seen in animals treated with APB during the normal period of orientation development and then allowed several months of development without treatment. These results suggest that patterns of neuronal activity carried in the separate ON- and OFF-center visual pathways are necessary for the development of orientation selectivity in visual cortical neurons of the ferret and that there is a critical period for this development. PMID- 10684894 TI - Properties of horizontal and vertical inputs to pyramidal cells in the superficial layers of the cat visual cortex. AB - The purpose of this study is to elucidate the integrative input mechanisms of pyramidal cells receiving horizontally projecting axon collaterals (horizontal projection) and vertical input from layer IV. We performed whole-cell recordings from pyramidal cells in layer II/III and focally activated other single pyramidal cells monosynaptically connected via long-distance horizontal (LH) projections (the distance between presynaptic and postsynaptic cells was 350-1200 micrometer) in slice preparations of the kitten primary visual cortex. In addition, presynaptic single fibers in layer IV (vertical input) and/or short-distance horizontal (SH) inputs from neighboring single pyramidal cells (distance within 100 micrometer) in layer II/III were activated. Unitary EPSPs evoked by the activation of LH and SH connections had smaller amplitude and larger coefficient of variation than those evoked by stimulating the vertical input. Paired-pulse stimulation of the LH and SH inputs caused the depression of the second EPSP, whereas that of vertical inputs caused either facilitation or depression of the second EPSP. The EPSPs evoked by simultaneous activation of LH and vertical inputs summated linearly at the resting membrane potential. However, the EPSPs evoked by stimulation of the two inputs were nonlinearly (supralinearly) summated when the postsynaptic membrane was depolarized to a certain level. Similar EPSP interaction was observed in response to simultaneous activation of the LH and SH inputs. PMID- 10684895 TI - Three levels of lateral inhibition: A space-time study of the retina of the tiger salamander. AB - The space-time patterns of activity generated across arrays of retinal neurons can provide a sensitive measurement of the effects of neural interactions underlying retinal activity. We measured the excitatory and inhibitory components associated with these patterns at each cellular level in the retina and further dissected inhibitory components pharmacologically. Using perforated and loose patch recording, we measured the voltages, currents, or spiking at 91 lateral positions covering approximately 2 mm in response to a flashed 300-microm-wide bar. First, we showed how the effect of well known lateral inhibition at the outer retina, mediated by horizontal cells, evolved in time to compress the spatial representation of the stimulus bar at ON and OFF bipolar cell bodies as well as horizontal cells. Second, we showed, for the first time, how GABA(C) receptor mediated amacrine cell feedback to bipolar terminals compresses the spatial representation of the stimulus bar at ON bipolar terminals over time. Third, we showed that a third spatiotemporal compression exists at the ganglion cell layer that is mediated by feedforward amacrine cells via GABA(A) receptors. These three inhibitory mechanisms, via three different receptor types, appear to compensate for the effects of lateral diffusion of activity attributable to dendritic spread and electrical coupling between retinal neurons. As a consequence, the width of the final representation at the ganglion cell level approximates the dimensions of the original stimulus bar. PMID- 10684896 TI - Dual serotonin (5-HT) projections to the nucleus accumbens core and shell: relation of the 5-HT transporter to amphetamine-induced neurotoxicity. AB - Dopamine release in the nucleus accumbens (NAc) has been implicated as mediating the rewarding effects of stimulant drugs; however, recent studies suggest that 5 HT release may also contribute. In an effort to assess the role of 5-HT in drug mediated reward, this study analyzed the serotonergic innervation of NAc using immunocytochemistry for 5-HT and the 5-HT transporter (SERT). We report that in control rats the NAc receives two distinct types of 5-HT axons that differ in regional distribution, morphology, and SERT expression. Most regions of the NAc are innervated by thin 5-HT axons that express SERT, but in the caudal NAc shell nearly all 5-HT axons lack SERT and have large spherical varicosities. Two weeks after methamphetamine or p-chloroamphetamine (PCA) treatment, most 5-HT axons in dorsal striatum and NAc have degenerated; however, the varicose axons in the shell appear intact. These drug-resistant 5-HT axons that lack SERT densely innervate the caudal one-third of the accumbens shell, the same location where dopamine axons are spared after methamphetamine. Moreover, 4 hr after PCA, the varicose axons in the caudal shell retain prominent stores of 5-HT, whereas 5-HT axons in the rest of the NAc are depleted of neurotransmitter. The results demonstrate that two functionally different 5-HT projections innervate separate regions of the NAc and that selective vulnerability to amphetamines may result from differential expression of SERT. We postulate that action potentials conducted from the raphe nuclei can release 5-HT throughout the NAc, whereas transporter-mediated release induced by stimulant drugs is more restricted and unlikely to occur in the caudal NAc shell. PMID- 10684897 TI - Interspike intervals, receptive fields, and information encoding in primary visual cortex. AB - In the primate primary visual cortex (V1), the significance of individual action potentials has been difficult to determine, particularly in light of the considerable trial-to-trial variability of responses to visual stimuli. We show here that the information conveyed by an action potential depends on the duration of the immediately preceding interspike interval (ISI). The interspike intervals can be grouped into several different classes on the basis of reproducible features in the interspike interval histograms. Spikes in different classes bear different relationships to the visual stimulus, both qualitatively (in terms of the average stimulus preceding each spike) and quantitatively (in terms of the amount of information encoded per spike and per second). Spikes preceded by very short intervals (3 msec or less) convey information most efficiently and contribute disproportionately to the overall receptive-field properties of the neuron. Overall, V1 neurons can transmit between 5 and 30 bits of information per second in response to rapidly varying, pseudorandom stimuli, with an efficiency of approximately 25%. Although some (but not all) of our results would be expected from neurons that use a firing-rate code to transmit information, the evidence suggests that visual neurons are well equipped to decode stimulus related information on the basis of relative spike timing and ISI duration. PMID- 10684898 TI - Conscious and unconscious processing of nonverbal predictability in Wernicke's area. AB - The association of nonverbal predictability and brain activation was examined using functional magnetic resonance imaging in humans. Participants regarded four squares displayed horizontally across a screen and counted the incidence of a particular color. A repeating spatial sequence with varying levels of predictability was embedded within a random color presentation. Both Wernicke's area and its right homolog displayed a negative correlation with temporal predictability, and this effect was independent of individuals' conscious awareness of the sequence. When individuals were made aware of the underlying sequential predictability, a widespread network of cortical regions displayed activity that correlated with the predictability. Conscious processing of predictability resulted in a positive correlation to activity in right prefrontal cortex but a negative correlation in posterior parietal cortex. These results suggest that conscious processing of predictability invokes a large-scale cortical network, but independently of awareness, Wernicke's area processes predictive events in time and may not be exclusively associated with language. PMID- 10684899 TI - Neuroactive steroid 3alpha-hydroxy-5alpha-pregnan-20-one modulates electrophysiological and behavioral actions of ethanol. AB - Neuroactive steroids are synthesized de novo in brain, yet their physiological significance remains elusive. We provide biochemical, electrophysiological, and behavioral evidence that several specific actions of alcohol (ethanol) are mediated by the neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha THP; allopregnanolone). Systemic alcohol administration elevates 3alpha, 5alpha THP levels in the cerebral cortex to pharmacologically relevant concentrations. The elevation of 3alpha,5alpha-THP is dose- and time-dependent. Furthermore, there is a significant correlation between 3alpha,5alpha-THP levels in cerebral cortex and the hypnotic effect of ethanol. Blockade of de novo biosynthesis of 5alpha-reduced steroids using the 5alpha-reductase inhibitor finasteride prevents several effects of ethanol. Pretreatment with finasteride causes no changes in baseline bicuculline-induced seizure threshold but reverses the anticonvulsant effect of ethanol. Finasteride pretreatment also reverses ethanol inhibition of spontaneous neural activity in medial septal/diagonal band of Broca neurons while having no direct effect on spontaneous firing rates. Thus, elevation of 3alpha,5alpha-THP levels by acute ethanol administration represents a novel mechanism of ethanol action as well as an important modulatory role for neurosteroids in the CNS. PMID- 10684900 TI - A proprioceptive role for an exteroceptive mechanoafferent neuron in Aplysia. AB - Afferent regulation of centrally generated activity is likely to be more complex than has been established. We show that a neuron that is an exteroceptor can also function as a proprioceptor. We study the Aplysia neuron B21. Previous data suggest that B21 functions as an exteroceptor during the radula closing/retraction phase of ingestive feeding. We show that the tissue innervated by B21, the subradula tissue (SRT), is innervated by a motor neuron (B66) and that B66-induced SRT contractions trigger centripetal spikes in B21. Thus, B21 is also a proprioceptor. To determine whether exteroceptive and proprioceptive activities occur during the same phase of ingestive feeding, we further characterize B66. We show that B66 stimulation does not close or retract the radula. Instead it opens it. Moreover, B66 is electrically coupled to other opening/protraction neurons. Finally, we elicit motor programs in semi-intact preparations and show that during radula opening/protraction we observe B66 activity, SRT contractions, and spikes in B21 that can be eliminated if B66 is indirectly hyperpolarized. B21 is, therefore, likely to act as an exteroceptor during one phase of ingestive feeding and as a proprioceptor during the antagonistic phase. Previous experiments have shown that centripetal spikes in B21 are only transmitted to one follower if they are "gated in" by depolarization. During ingestive programs B21 is centrally depolarized during closing/retraction, but it is not depolarized during opening/protraction. We sought to determine whether there are other followers that receive B21 input when it is not centrally depolarized. We found one such cell. Moreover, we found that stimulation of B21 during radula opening/protraction significantly decreases the duration of this phase of behavior. Thus, proprioceptive activity in B21 is likely to have an impact on motor programs. PMID- 10684901 TI - Laminin degradation by plasmin regulates long-term potentiation. AB - Plasmin is converted from its zymogen plasminogen by tissue type or urokinase type plasminogen activator (PA) and degrades many components of the extracellular matrix (ECM). To explore the possibility that the PA-plasmin system regulates synaptic plasticity, we investigated the effect of plasmin on degradation of ECM and synaptic plasticity by using organotypic hippocampal cultures. High-frequency stimulation produced long-term potentiation (LTP) in control slices, whereas the potentiation was induced but not maintained in slices pretreated with 100 nM plasmin for 6 hr. The baseline synaptic responses were not affected by pretreatment with plasmin. The impairment of LTP maintenance was not observed in slices pretreated with 100 nM plasmin for 6 hr, washed, and then cultured for 24 48 hr in the absence of plasmin. To identify substrates of plasmin, the expression of three major components of ECM, laminin, fibronectin, and type IV collagen, was investigated by immunofluorescence imaging. The three ECM components were widely distributed in the hippocampus, and only laminin was degraded by plasmin pretreatment. The expression level of laminin returned to normal levels when the slices were cultured for 24-48 hr after washout of plasmin. Furthermore, preincubation with anti-laminin antibodies prevented both the degradation of laminin and the impairment of LTP maintenance by plasmin. These results suggest that the laminin-mediated cell-ECM interaction may be necessary for the maintenance of LTP. PMID- 10684902 TI - Long-lasting depolarizations in mitral cells of the rat olfactory bulb. AB - We investigated the mechanisms of long-lasting depolarizing potentials (LLDs) generated in mitral cells with whole-cell patch recordings in the rat olfactory bulb slice. LLDs occur spontaneously and are evoked by either orthodromic stimulation of the olfactory nerve or antidromic stimulation of mitral and tufted (M/T) cells. LLDs are followed by a long refractory period, limiting LLD generation to approximately 1 Hz. LLD production does not appear to involve either intrinsic voltage-activated or metabotropic mechanisms. The initiation of LLDs requires activation of non-NMDA but not NMDA receptors. Dual recordings from the apical dendrites and somata of mitral cells show that LLDs are generated in the distal portion of the apical dendrite, most likely in the glomerulus. The rising phase of LLDs shows characteristics of polyneuronal input, including a high variability and sensitivity to charge screening. Paired recordings from adjacent mitral cells suggest that LLDs occur synchronously only in cells whose apical dendrites ramify in the same glomerulus. These findings suggest that LLDs involve recurrent, intraglomerular dendrodendritic interactions among M/T cells. PMID- 10684903 TI - Modulation of presynaptic action potential kinetics underlies synaptic facilitation of type B photoreceptors after associative conditioning in Hermissenda. AB - Descriptions of conditioned response generation in Hermissenda stipulate that the synaptic interaction between type B and A photoreceptors should be enhanced after associative pairings of light and rotation. Although evidence from several laboratories has confirmed this assumption, the mechanism underlying this synaptic facilitation has not been elucidated. Here we report that in vitro conditioning (i.e., light paired with stimulation of vestibular hair cells) modifies the kinetics of presynaptic action potentials in the B photoreceptor in a manner sufficient to account for this synaptic facilitation. After paired training, we observed an increase in the duration of evoked action potentials and a decrease in the amplitude of the spike afterhyperpolarization in the B-cell. As previously reported, paired training also enhanced the excitability (i.e., input resistance and evoked spike rate) of the B photoreceptor. In a second experiment, simultaneous recordings were made in type B and A photoreceptors, and paired training was found to produce an increase in the amplitude of the IPSP in the A photoreceptor in response to an evoked spike in the B-cell. Importantly, there was no change in the initial slope of the postsynaptic IPSP in the A photoreceptor, suggesting that spike duration-independent mechanisms of neurotransmitter exocytosis or postsynaptic receptor sensitivity did not contribute to the observed synaptic facilitation. Perfusion of 4-aminopyridine (4 AP) mimicked a known effect of behavioral conditioning in that it specifically reduced the amplitude of the transient voltage-dependent K(+) current (I(A)) in the B-cell, but in addition, produced action potential broadening and synaptic facilitation that was analogous to that observed after in vitro conditioning. Finally, the effect of 4-AP on B-cell action potentials and on the postsynaptic IPSP in the A-cell was occluded by previous paired (but not unpaired) training, suggesting that the prolongation of the B-cell action potential by a reduction of I(A) was sufficient to account for the observed synaptic facilitation. The occlusion of the effects of 4-AP by paired training was not attributable to a saturation of the capacity of the B-cell for transmitter exocytosis, because it was observed that tetraethylammonium (TEA)-induced inhibition of the delayed voltage-dependent K(+) current induced both spike broadening and synaptic facilitation regardless of training history. Collectively, these results demonstrate that training-induced facilitation at B-cell synapses is attributable to the effects of a reduction of a presynaptic K(+) conductance on action potential kinetics and suggest another critical similarity between the cellular basis for learning in Hermissenda and other invertebrate systems. PMID- 10684904 TI - Peptide cotransmitter release from motorneuron B16 in aplysia californica: costorage, corelease, and functional implications. AB - Many neurons contain multiple peptide cotransmitters in addition to their classical transmitters. We are using the accessory radula closer neuromuscular system of Aplysia, which participates in feeding in these animals, to define the possible consequences of multiple modulators converging on single targets. How these modulators are released onto their targets is of critical importance in understanding the outcomes of their modulatory actions and their physiological role. Here we provide direct evidence that the partially antagonistic families of modulatory peptides, the myomodulins and buccalins, synthesized by motorneuron B16 are costored and coreleased in fixed ratios. We show that this release is calcium-dependent and independent of muscle contraction. Furthermore, we show that peptide release is initiated at the low end of the physiological range of motorneuron firing frequency and that it increases with increasing motorneuron firing frequency. The coordination of peptide release with the normal operating range of a neuron may be a general phenomenon and suggests that the release of peptide cotransmitters may exhibit similar types of regulation and plasticity as have been observed for classical transmitters. Stimulation paradigms that increase muscle contraction amplitude or frequency also increase peptide release from motor neuron B16. The net effect of the modulatory peptide cotransmitters released from motorneuron B16 would be to increase relaxation rate and therefore allow more frequent and/or larger contractions to occur without increased resistance to antagonist muscles. The end result of this modulation could be to maximize the efficiency of feeding. PMID- 10684905 TI - Fine structure of parvocellular receptive fields in the primate fovea revealed by laser interferometry. AB - Optical blurring in the eye prevents conventional physiological techniques from revealing the fine structure of the small parvocellular receptive fields in the primate fovea in vivo. We explored the organization of receptive fields in macaque parvocellular lateral geniculate nucleus cells by using sinusoidal interference fringes formed directly on the retina to measure spatial frequency tuning at different orientations. Most parvocellular cells in and near the fovea respond reliably to spatial frequencies up to and beyond 100 cycles/ degrees of visual angle, implying center input arising mainly from a single cone. Temporal frequency and contrast response characteristics were also measured at spatial frequencies up to 130 cycles/degrees. We compared our spatial frequency data with the frequency responses of model receptive fields that estimate the number, configuration, and weights of cones that feed the center and surround. On the basis of these comparisons, we infer possible underlying circuits. Most cells had irregular spatial frequency-response curves that imply center input from more than one cone. The measured responses are consistent with a single cone center together with weak input from nearby cones. By exposing a fine structure that cannot be discerned by conventional techniques, interferometry allows functional measurements of the early neural mechanisms in spatial vision. PMID- 10684906 TI - Control of synaptic depression by glutamate transporters. AB - The role of glutamate transporters in the regulation of synaptic depression was examined in the avian nucleus magnocellularis. Repetitive stimulation of presynaptic auditory nerve fibers resulted in acute depression of EPSCs. Pharmacological blockade of glutamate transport in glial cells enhanced residual glutamate in the synaptic cleft and markedly increased the extent of depression at stimulus frequencies above 20 Hz via a postsynaptic mechanism. Glutamate pyruvate transaminase, a glutamate scavenger, accelerated the decay of the EPSC and reduced synaptic depression, indicating that transporters are not completely effective in rapid removal of glutamate. Regulation of residual transmitter by glia may thus serve to control synaptic strength in a frequency-dependent manner. PMID- 10684907 TI - Hypothalamic-pituitary-adrenal dysfunction in Apoe(-/-) mice: possible role in behavioral and metabolic alterations. AB - Several neurological diseases are frequently accompanied by dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis. The HPA axis regulates the secretion of glucocorticoids (GCs), which play important roles in diverse brain functions, including cognition, emotion, and feeding. Under physiological conditions, GCs are adaptive and beneficial; however, prolonged elevations in GC levels may contribute to neurodegeneration and brain dysfunction. In the current study, we demonstrate that apolipoprotein E (apoE) deficiency results in age-dependent dysregulation of the HPA axis through a mechanism affecting primarily the adrenal gland. Apoe(-/-) mice, which develop neurodegenerative alterations as they age, had an age-dependent increase in basal adrenal corticosterone content and abnormally increased plasma corticosterone levels after restraint stress, whereas their plasma and pituitary adrenocorticotropin levels were either unchanged or lower than those in controls. HPA axis dysregulation was associated with behavioral and metabolic alterations. When anxiety levels were assessed in the elevated plus maze, Apoe(-/-) mice showed more anxiety than wild-type controls. Apoe(-/-) mice also showed reduced activity in the open field. Finally, Apoe(-/-) mice showed age-dependent increases in food and water intake, stomach and body weights, and decreases in brown and white adipose tissues. These results support a key role for apoE in the tonic inhibition of steroidogenesis and HPA axis activity and have important implications for the behavioral analysis of Apoe(-/-) mice. PMID- 10684908 TI - Selectivity for complex shapes in primate visual area V2. AB - To explore the role of visual area V2 in shape analysis, we studied the responses of neurons in area V2 of the alert macaque using a set of 128 grating and geometric line stimuli that varied in their shape characteristics and geometric complexity. Simple stimuli included oriented bars and sinusoidal gratings; complex stimuli included angles, arcs, circles, and intersecting lines, plus hyperbolic and polar gratings. We found that most V2 cells responded well to at least some of the complex stimuli, and in many V2 cells the most effective complex stimulus elicited a significantly larger response than the most effective bar or sinusoid. Approximately one-third of the V2 cells showed significant differential responsiveness to various complex shape characteristics, and many were also selective for the orientation, size, and/or spatial frequency of the preferred shape. These results indicate that V2 cells explicitly represent complex shape information and suggest specific types of higher order visual information that V2 cells extract from visual scenes. PMID- 10684909 TI - Distinct sites of opiate reward and aversion within the midbrain identified using a herpes simplex virus vector expressing GluR1. AB - Repeated administration of morphine increases expression of GluR1 (an AMPA glutamate receptor subunit) in the ventral tegmental area (VTA) of the midbrain, an important neural substrate for the rewarding actions of morphine. Microinjections of a herpes simplex virus (HSV) vector that causes local overexpression of GluR1 (HSV-GluR1) into the VTA can enhance the ability of morphine to establish conditioned place preferences, suggesting that altered GluR1 expression in this region is directly associated with changes in the rewarding efficacy of morphine. We now report that in rats given HSV-GluR1 directly into the VTA, morphine is most rewarding when maximal transgene expression is in the rostral VTA, whereas morphine is aversive when maximal transgene expression is in the caudal VTA. Dual-labeling immunohistochemistry shows that this difference cannot be explained by a different fraction of dopaminergic neurons infected in the rostral versus caudal VTA. No such anatomical specificity is seen in rats given VTA microinjections of HSV-LacZ, a vector expressing a control protein (-galactosidase). These results suggest that distinct substrates within the VTA itself differentially contribute to the rewarding and aversive properties of opiates. PMID- 10684910 TI - Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. AB - BACKGROUND: Cytokeratins are specific markers of epithelial cancer cells in bone marrow. We assessed the influence of cytokeratin-positive micrometastases in the bone marrow on the prognosis of women with breast cancer. METHODS: We obtained bone marrow aspirates from both upper iliac crests of 552 patients with stage I, II, or III breast cancer who underwent complete resection of the tumor and 191 patients with nonmalignant disease. The specimens were stained with the monoclonal antibody A45-B/B3, which binds to an antigen on cytokeratins. The median follow-up was 38 months (range, 10 to 70). The primary end point was survival. RESULTS: Cytokeratin-positive cells were detected in the bone marrow specimens of 2 of the 191 control patients with nonmalignant conditions (1 percent) and 199 of the 552 patients with breast cancer (36 percent). The presence of occult metastatic cells in bone marrow was unrelated to the presence or absence of lymph-node metastasis (P=0.13). After four years of follow-up, the presence of micrometastases in bone marrow was associated with the occurrence of clinically overt distant metastasis and death from cancer-related causes (P<0.001), but not with locoregional relapse (P=0.77). Of 199 patients with occult metastatic cells, 49 died of cancer, whereas of 353 patients without such cells, 22 died of cancer-related causes (P<0.001). Among the 301 women without lymph-node metastases, 14 of the 100 with bone marrow micrometastases died of cancer-related causes, as did 2 of the 201 without bone marrow micrometastases (P<0.001). The presence of occult metastatic cells in bone marrow, as compared with their absence, was an independent prognostic indicator of the risk of death from cancer (relative risk, 4.17; 95 percent confidence interval, 2.51 to 6.94; P<0.001), after adjustment for the use of systemic adjuvant chemotherapy. CONCLUSIONS: The presence of occult cytokeratin-positive metastatic cells in bone marrow increases the risk of relapse in patients with stage I, II, or III breast cancer. PMID- 10684911 TI - Metronidazole to prevent preterm delivery in pregnant women with asymptomatic bacterial vaginosis. National Institute of Child Health and Human Development Network of Maternal-Fetal Medicine Units. AB - BACKGROUND: Bacterial vaginosis has been associated with preterm birth. In clinical trials, the treatment of bacterial vaginosis in pregnant women who previously had a preterm delivery reduced the risk of recurrence. METHODS: To determine whether treating women in a general obstetrical population who have asymptomatic bacterial vaginosis (as diagnosed on the basis of vaginal Gram's staining and pH) prevents preterm delivery, we randomly assigned 1953 women who were 16 to less than 24 weeks pregnant to receive two 2-g doses of metronidazole or placebo. The diagnostic studies were repeated and a second treatment was administered to all the women at 24 to less than 30 weeks' gestation. The primary outcome was the rate of delivery before 37 weeks' gestation. RESULTS: Bacterial vaginosis resolved in 657 of 845 women who had follow-up Gram's staining in the metronidazole group (77.8 percent) and 321 of 859 women in the placebo group (37.4 percent). Data on the time and characteristics of delivery were available for 953 women in the metronidazole group and 966 in the placebo group. Preterm delivery occurred in 116 women in the metronidazole group (12.2 percent) and 121 women in the placebo group (12.5 percent) (relative risk, 1.0; 95 percent confidence interval, 0.8 to 1.2). Treatment did not prevent preterm deliveries that resulted from spontaneous labor (5.1 percent in the metronidazole group vs. 5.7 percent in the placebo group) or spontaneous rupture of the membranes (4.2 percent vs. 3.7 percent), nor did it prevent delivery before 32 weeks (2.3 percent vs. 2.7 percent). Treatment with metronidazole did not reduce the occurrence of preterm labor, intraamniotic or postpartum infections, neonatal sepsis, or admission of the infant to the neonatal intensive care unit. CONCLUSIONS: The treatment of asymptomatic bacterial vaginosis in pregnant women does not reduce the occurrence of preterm delivery or other adverse perinatal outcomes. PMID- 10684912 TI - Orthostatic intolerance and tachycardia associated with norepinephrine transporter deficiency. AB - BACKGROUND: Orthostatic intolerance is a syndrome characterized by lightheadedness, fatigue, altered mentation, and syncope and associated with postural tachycardia and plasma norepinephrine concentrations that are disproportionately high in relation to sympathetic outflow. We tested the hypothesis that impaired functioning of the norepinephrine transporter contributes to the pathophysiologic mechanism of orthostatic intolerance. METHODS: In a patient with orthostatic intolerance and her relatives, we measured postural blood pressure, heart rate, plasma catecholamines, and systemic norepinephrine spillover and clearance, and we sequenced the norepinephrine transporter gene and evaluated its function. RESULTS: The patient had a high mean plasma norepinephrine concentration while standing, as compared with the mean (+/ SD) concentration in normal subjects (923 vs. 439+/-129 pg per milliliter [5.46 vs. 2.59+/-0.76 nmol per liter]), reduced systemic norepinephrine clearance (1.56 vs. 2.42+/-0.71 liters per minute), impairment in the increase in the plasma norepinephrine concentration after the administration of tyramine (12 vs. 56+/-63 pg per milliliter [0.07 vs. 0.33+/-0.37 pmol per liter]), and a disproportionate increase in the concentration of plasma norepinephrine relative to that of dihydroxyphenylglycol. Analysis of the norepinephrine-transporter gene revealed that the proband was heterozygous for a mutation in exon 9 (encoding a change from guanine to cytosine at position 237) that resulted in more than a 98 percent loss of function as compared with that of the wild-type gene. Impairment of synaptic norepinephrine clearance may result in a syndrome characterized by excessive sympathetic activation in response to physiologic stimuli. The mutant allele in the proband's family segregated with the postural heart rate and abnormal plasma catecholamine homeostasis. CONCLUSIONS: Genetic or acquired deficits in norepinephrine inactivation may underlie hyperadrenergic states that lead to orthostatic intolerance. PMID- 10684913 TI - Images in clinical medicine. Pituitary apoplexy. PMID- 10684914 TI - Clinical problems with the performance of euthanasia and physician-assisted suicide in The Netherlands. AB - BACKGROUND AND METHODS: The characteristics and frequency of clinical problems with the performance of euthanasia and physician-assisted suicide are uncertain. We analyzed data from two studies of euthanasia and physician-assisted suicide in The Netherlands (one conducted in 1990 and 1991 and the other in 1995 and 1996), with a total of 649 cases. We categorized clinical problems as technical problems, such as difficulty inserting an intravenous line; complications, such as myoclonus or vomiting; or problems with completion, such as a longer-than expected interval between the administration of medications and death. RESULTS: In 114 cases, the physician's intention was to provide assistance with suicide, and in 535, the intention was to perform euthanasia. Problems of any type were more frequent in cases of assisted suicide than in cases of euthanasia. Complications occurred in 7 percent of cases of assisted suicide, and problems with completion (a longer-than-expected time to death, failure to induce coma, or induction of coma followed by awakening of the patient) occurred in 16 percent of the cases; complications and problems with completion occurred in 3 percent and 6 percent of cases of euthanasia, respectively. The physician decided to administer a lethal medication in 21 of the cases of assisted suicide (18 percent), which thus became cases of euthanasia. The reasons for this decision included problems with completion (in 12 cases) and the inability of the patient to take all the medications (in 5). CONCLUSIONS: There may be clinical problems with the performance of euthanasia and physician-assisted suicide. In The Netherlands, physicians who intend to provide assistance with suicide sometimes end up administering a lethal medication themselves because of the patient's inability to take the medication or because of problems with the completion of physician assisted suicide. PMID- 10684915 TI - Physicians' experiences with the Oregon Death with Dignity Act. AB - BACKGROUND: Physician-assisted suicide was legalized in Oregon in October 1997. There are data on patients who have received prescriptions for lethal medications and died after taking the medications. There is little information, however, on physicians' experiences with requests for assistance with suicide. METHODS: Between February and August 1999, we mailed a questionnaire to physicians who were eligible to prescribe lethal medications under the Oregon Death with Dignity Act. RESULTS: Of 4053 eligible physicians, 2649 (65 percent) returned the survey. Of the respondents, 144 (5 percent) had received a total of 221 requests for prescriptions for lethal medications since October 1997. We received information on the outcome in 165 patients (complete information for 143 patients and partial for on an additional 22). The mean age of the patients was 68 years; 76 percent had an estimated life expectancy of less than six months. Thirty-five percent requested a prescription from another physician. Twenty-nine patients (18 percent) received prescriptions, and 17 (10 percent) died from administering the prescribed medication. Twenty percent of the patients had symptoms of depression; none of these patients received a prescription for a lethal medication. In the case of 68 patients, including 11 who received prescriptions and 8 who died by taking the prescribed medication, the physician implemented at least one substantive palliative intervention, such as control of pain or other symptoms, referral to a hospice program, a consultation, or a trial of antidepressant medication. Forty-six percent of the patients for whom substantive interventions were made changed their minds about assisted suicide, as compared with 15 percent of those for whom no substantive interventions were made (P<0.001). CONCLUSIONS: Our data indicate that in Oregon, physicians grant about 1 in 6 requests for a prescription for a lethal medication and that 1 in 10 requests actually result in suicide. Substantive palliative interventions lead some--but not all--patients to change their minds about assisted suicide. PMID- 10684916 TI - Assessing the risk of breast cancer. PMID- 10684917 TI - Case Records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 6-2000. Hemoptysis in a 20-year-old man with multiple pulmonary nodules. PMID- 10684918 TI - Approaches to breast-cancer staging. PMID- 10684919 TI - Antibiotics for the prevention of preterm birth. PMID- 10684920 TI - Physician-assisted suicide and euthanasia in practice. PMID- 10684921 TI - Legalized physician-assisted suicide in Oregon--the second year. AB - BACKGROUND AND METHODS: In 1997, Oregon legalized physician-assisted suicide. We have previously reported data on terminally ill Oregon residents who received prescriptions for lethal medications under the Oregon Death with Dignity Act and who died in 1998. We now report similar data for 1999, obtained from physicians' reports, death certificates, and interviews with physicians. We also report data from interviews with family members. RESULTS: Information on 33 persons who received prescriptions for lethal medications in 1999 was reported to the Oregon Health Division; 26 died after taking the lethal medications, 5 died from their underlying illnesses, and 2 were alive as of January 1, 2000. One additional patient, who received a prescription in 1998, died after taking the medication in 1999. Thus, 27 patients died after ingesting lethal medications in 1999 (9 per 10,000 deaths in Oregon), as compared with 16 patients in 1998 (6 per 10,000). The median age of the 27 patients who died in 1999 after taking lethal medications was 71 years. The most frequent underlying illnesses were cancer (in 17 patients), amyotrophic lateral sclerosis (in 4), and chronic obstructive pulmonary disease (in 4). All 27 patients had health insurance, 21 were receiving hospice care, and 13 were college graduates. According to both physicians and family members, patients requested assistance with suicide for several reasons, including loss of autonomy, loss of control of bodily functions, an inability to participate in activities that make life enjoyable, and a determination to control the manner of death. CONCLUSIONS: In the second as compared with the first year of legalized physician-assisted suicide in Oregon, the number of patients who died after ingesting lethal medications increased, but it remained small in relation to the total number of persons in Oregon who died. Patients who request assistance with suicide appear to be motivated by several factors, including loss of autonomy and a determination to control the way in which they die. PMID- 10684923 TI - Intronic GIY-YIG endonuclease gene in the mitochondrial genome of Podospora curvicolla: evidence for mobility. AB - Endonuclease genes encoded in invasive introns are themselves supposed to be mobile elements which, during evolution, have colonized pre-existing introns converting them into invasive elements. This hypothesis is supported by numerous data concerning the LAGLI-DADG subclass of intronic endonucleases. Less is known about the GIY-YIG ORFs which constitute another family of endonucleases. In this paper we describe the presence of one optional GIY-YIG ORF in the second intron of the mitochondrial cytochrome b gene in the fungus Podospora curvicolla. We show that this GIY-YIG ORF is efficiently transferred from an ORF-containing intron to an ORF-less allele. We also show that the products of both the GIY-YIG ORF and the non-canonical LAGLI-DADG-GIY-YIG ORF, which is generated by its integration, have endonuclease activities which recognize and cut the insertion site of the optional sequence. This constitutes the first direct evidence for potential mobility of an intronic GIY-YIG endonuclease. We discuss the role that such a mobile sequence could have played during evolution. PMID- 10684924 TI - Cold shock induces the insertion of a cryptic exon in the neurofibromatosis type 1 (NF1) mRNA. AB - Alternative splicing is a regulatory process of gene expression based on the flexibility in the selection of splice sites. In this manuscript we present the characterisation of an alternative splicing of the NF1 pre-mRNA induced by cold shock conditions. We demonstrate that the accuracy of the splicing mechanism was perturbed after keeping samples for a short period of time at room temperature, resulting in the insertion of a 31-bp cryptic exon between exons 4a and 4b of the NF1 mRNA. This alternative splicing is not cell type specific and is not induced by other stress conditions such as heat shock or hyper-osmolarity. The alternative spliced mRNA is efficiently transported to the cytoplasm and it is proven to belong to the poly A(+)mRNA fraction. Previous misleading interpretations about this transcript, together with our finding relating its presence to cold shock and not to the NF1 disease, strongly indicate that this phenomenon should be taken into account in genetic testing when RNA methodology is used for mutation detection. This is the first description of an alternative splicing induced by cold shock in a human pre-mRNA and should provide further insights into the factors that control alternative splicing. PMID- 10684925 TI - Interaction of the yeast DExH-box RNA helicase prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing. AB - Using site-specific incorporation of the photo-chemical cross-linking reagent 4 thiouridine, we demonstrate the previously unknown association of two proteins with yeast 3' splice sites. One of these is an unidentified approximately 122 kDa protein that cross-links to 3' splice sites during formation of the pre- spliceosome. The other factor is the DExH-box RNA helicase, Prp22p. With substrates functional in the second step of splicing, only very weak cross linking of Prp22p to intron sequences at the 3' splice site is observed. In contrast, substrates blocked at the second step exhibit strong cross-linking of Prp22 to intron sequences at the 3' splice site, but not to adjacent exon sequences. In vitro reconstitution experiments also show that the association of Prp22p with intron sequences at the 3' splice site is dependent on Prp16p and does not persist when release of mature mRNA from the spliceosome is blocked. Taken together, these results suggest that the 3' splice site of yeast introns is contacted much earlier than previously envisioned by a protein of approximately 120 kDa, and that a transient association of Prp22p with the 3' splice site occurs between the first and second catalytic steps. PMID- 10684926 TI - The solution structure of [d(CGC)r(aaa)d(TTTGCG)](2): hybrid junctions flanked by DNA duplexes. AB - The solution structure and hydration of the chimeric duplex [d(CGC)r(aaa)d(TTTGCG)](2), in which the central hybrid segment is flanked by DNA duplexes at both ends, was determined using two-dimensional NMR, simulated annealing and restrained molecular dynamics. The solution structure of this chimeric duplex differs from the previously determined X-ray structure of the analogous B-DNA duplex [d(CGCAAATTTGCG)](2)as well as NMR structure of the analogous A-RNA duplex [r(cgcaaauuugcg)](2). Long-lived water molecules with correlation time tau(c)longer than 0.3 ns were found close to the RNA adenine H2 and H1' protons in the hybrid segment. A possible long-lived water molecule was also detected close to the methyl group of 7T in the RNA-DNA junction but not with the other two thymines (8T and 9T). This result correlates with the structural studies that only DNA residue 7T in the RNA-DNA junction adopts an O4' endo sugar conformation, while the other DNA residues including 3C in the DNA-RNA junction, adopt C1'-exo or C2'-endo conformations. The exchange rates for RNA C2' OH were found to be approximately 5-20 s(-1). This slow exchange rate may be due to the narrow minor groove width of [d(CGC)r(aaa)d(TTTGCG)](2), which may trap the water molecules and restrict the dynamic motion of hydroxyl protons. The minor groove width of [d(CGC)r(aaa)d(TTTGCG)](2)is wider than its B-DNA analog but narrower than that of the A-RNA analog. It was further confirmed by its titration with the minor groove binding drug distamycin. A possible 2:1 binding mode was found by the titration experiments, suggesting that this chimeric duplex contains a wider minor groove than its B-DNA analog but still narrow enough to hold two distamycin molecules. These distinct structural features and hydration patterns of this chimeric duplex provide a molecular basis for further understanding the structure and recognition of DNA. RNA hybrid and chimeric duplexes. PMID- 10684922 TI - Survey and summary: transcription by RNA polymerases I and III. AB - The task of transcribing nuclear genes is shared between three RNA polymerases in eukaryotes: RNA polymerase (pol) I synthesizes the large rRNA, pol II synthesizes mRNA and pol III synthesizes tRNA and 5S rRNA. Although pol II has received most attention, pol I and pol III are together responsible for the bulk of transcriptional activity. This survey will summarise what is known about the process of transcription by pol I and pol III, how it happens and the proteins involved. Attention will be drawn to the similarities between the three nuclear RNA polymerase systems and also to their differences. PMID- 10684927 TI - Interactions of the human, rat, Saccharomyces cerevisiae and Escherichia coli 3 methyladenine-DNA glycosylases with DNA containing dIMP residues. AB - In DNA, the deamination of dAMP generates 2'-deoxy-inosine 5'-monophosphate (dIMP). Hypoxanthine (HX) residues are mutagenic since they give rise to A.T- >G.C transition. They are excised, although with different efficiencies, by an activity of the 3-methyl-adenine (3-meAde)-DNA glycosylases from Escherichia coli (AlkA protein), human cells (ANPG protein), rat cells (APDG protein) and yeast (MAG protein). Comparison of the kinetic constants for the excision of HX residues by the four enzymes shows that the E.coli and yeast enzymes are quite inefficient, whereas for the ANPG and the APDG proteins they repair the HX residues with an efficiency comparable to that of alkylated bases, which are believed to be the primary substrates of these DNA glycosylases. Since the use of various substrates to monitor the activity of HX-DNA glycosylases has generated conflicting results, the efficacy of the four 3-meAde-DNA glycosylases of different origin was compared using three different substrates. Moreover, using oligo-nucleotides containing a single dIMP residue, we investigated a putative sequence specificity of the enzymes involving the bases next to the HX residue. We found up to 2-5-fold difference in the rates of HX excision between the various sequences of the oligonucleotides studied. When the dIMP residue was placed opposite to each of the four bases, a preferential recognition of dI:T over dI:dG, dI:dC and dI:dA mismatches was observed for both human (ANPG) and E.coli (AlkA) proteins. At variance, the yeast MAG protein removed more efficiently HX from a dI:dG over dI:dC, dI:T and dI:dA mismatches. PMID- 10684928 TI - Effects of RNA secondary structure on cellular antisense activity. AB - The secondary and tertiary structures of a mRNA are known to effect hybridization efficiency and potency of antisense oligonucleotides in vitro. Additional factors including oligonucleotide stability and cellular uptake are also thought to contribute to antisense potency in vivo. Each of these factors can be affected by the sequence of the oligonucleotide. Although mRNA structure is presumed to be a critical determinant of antisense activity in cells, to date little direct experimental evidence has addressed the significance of structure. In order to determine the importance of mRNA structure on antisense activity, oligonucleotide target sites were cloned into a luciferase reporter gene along with adjoining sequence to form known structures. This allowed us to study the effect of target secondary structure on oligonucleotide binding in the cellular environment without changing the sequence of the oligonucleotide. Our results show that structure does play a significant role in determining oligonucleotide efficacy in vivo. We also show that potency of oligonucleotides can be improved by altering chemistry to increase affinity for the mRNA target even in a region that is highly structured. PMID- 10684929 TI - Characterisation of the U83 and U84 small nucleolar RNAs: two novel 2'-O-ribose methylation guide RNAs that lack complementarities to ribosomal RNAs. AB - In eukaryotic cells, the site-specific 2'- O -ribose methylation of ribosomal RNAs (rRNAs) and the U6 spliceosomal small nuclear RNA (snRNA) is directed by small nucleolar RNAs (snoRNAs). The C and D box-containing 2'- O -methylation guide snoRNAs select the correct substrate nucleotide through formation of a long 10-21 bp interaction with the target rRNA and U6 snRNA sequences. Here, we report on the characterisation of two novel mammalian C/D box snoRNAs, called U83 and U84, that contain all the elements that are essential for accumulation and function of 2'- O -methylation guide snoRNAs. However, in contrast to all of the known 2'- O -methylation guide RNAs, the human, mouse and pig U83 and U84 snoRNAs feature no antisense elements complementary to rRNA or U6 snRNA sequences. The human U83 and U84 snoRNAs are not associated with maturing nucleolar pre ribosomal particles, suggesting that they do not function in rRNA biogenesis. Since artificial substrate RNAs complementary to the evolutionarily conserved putative substrate recognition motifs of the U83 and U84 snoRNAs were correctly 2'- O -methylated in the nucleolus of mouse cells, we suggest that the new snoRNAs act as 2'- O -methylation guides for cellular RNAs other then rRNAs and the U6 snRNA. PMID- 10684930 TI - Identification of human MutY homolog (hMYH) as a repair enzyme for 2 hydroxyadenine in DNA and detection of multiple forms of hMYH located in nuclei and mitochondria. AB - An enzyme activity introducing an alkali-labile site at 2-hydroxyadenine (2-OH-A) in double-stranded oligonucleotides was detected in nuclear extracts of Jurkat cells. This activity co-eluted with activities toward adenine paired with guanine and 8-oxo-7,8-dihydroguanine (8-oxoG) as a single peak corresponding to a 55 kDa molecular mass on gel filtration chromatography. Further co-purification was then done. Western blotting revealed that these activities also co-purified with a 52 kDa polypeptide which reacted with antibodies against human MYH (anti-hMYH). Recombinant hMYH has essentially similar activities to the partially purified enzyme. Thus, hMYH is likely to possess both adenine and 2-OH-A DNA glycosylase activities. In nuclear extracts from Jurkat cells, a 52 kDa polypeptide was detected with a small amount of 53 kDa polypeptide, while in mitochondrial extracts a 57 kDa polypeptide was detected using anti-hMYH. With amplification of the 5'-regions of the hMYH cDNA, 10 forms of hMYH transcripts were identified and subgrouped into three types, each with a unique 5' sequence. These hMYH transcripts are likely to encode multiple authentic hMYH polypeptides including the 52, 53 and 57 kDa polypeptides detected in Jurkat cells. PMID- 10684931 TI - Assembly of archaeal signal recognition particle from recombinant components. AB - Signal recognition particle (SRP) takes part in protein targeting and secretion in all organisms. Searches for components of archaeal SRP in primary databases and completed genomes indicated that archaea possess only homologs of SRP RNA, and proteins SRP19 and SRP54. A recombinant SRP was assembled from cloned, expressed and purified components of the hyperthermophilic archaeon Archaeoglobus fulgidus. Recombinant Af-SRP54 associated with the signal peptide of bovine pre prolactin translated in vitro. As in mammalian SRP, Af-SRP54 binding to Af-SRP RNA required protein Af-SRP19, although notable amounts bound in absence of Af SRP19. Archaeoglobus fulgidus SRP proteins also bound to full-length SRP RNA of the archaeon Methanococcus jannaschii, to eukaryotic human SRP RNA, and to truncated versions which corresponded to the large domain of SRP. Dependence on SRP19 was most pronounced with components from the same species. Reconstitutions with heterologous components revealed a significant potential of human SRP proteins to bind to archaeal SRP RNAs. Surprisingly, M.jannaschii SRP RNA bound to human SRP54M quantitatively in the absence of SRP19. This is the first report of reconstitution of an archaeal SRP from recombinantly expressed purified components. The results highlight structural and functional conservation of SRP assembly between archaea and eucarya. PMID- 10684932 TI - Modified constructs of the tRNA TPsiC domain to probe substrate conformational requirements of m(1)A(58) and m(5)U(54) tRNA methyltransferases. AB - The TPsiC stem and loop (TSL) of tRNA contains highly conserved nucleoside modifications, m(5)C(49), T(54), Psi(55)and m(1)A(58). U(54)is methylated to m(5)U (T) by m(5)U(54)methyltransferase (RUMT); A(58)is methylated to m(1)A by m(1)A(58)tRNA methyltransferase (RAMT). RUMT recognizes and methylates a minimal TSL heptadecamer and RAMT has previously been reported to recognize and methylate the 3'-half of the tRNA molecule. We report that RAMT can recognize and methylate a TSL heptadecamer. To better understand the sensitivity of RAMT and RUMT to TSL conformation, we have designed and synthesized variously modified TSL constructs with altered local conformations and stabilities. TSLs were synthesized with natural modifications (T(54)and Psi(55)), naturally occurring modifications at unnatural positions (m(5)C(60)), altered sugar puckers (dU(54)and/or dU(55)) or with disrupted U-turn interactions (m(1)Psi(55)or m(1)m(3)Psi(55)). The unmodified heptadecamer TSL was a substrate of both RAMT and RUMT. The presence of T(54)increased thermal stability of the TSL and dramatically reduced RAMT activity toward the substrate. Local conformation around U(54)was found to be an important determinant for the activities of both RAMT and RUMT. PMID- 10684933 TI - Characterization of a cis-acting regulatory element in the protein coding region of thymidylate synthase mRNA. AB - Thymidylate synthase (TS) functions as an RNA-binding protein by interacting with two different sequences on its own mRNA. One site is located in the 5'-upstream region of human TS mRNA while the second site is located within the protein coding region corresponding to nt 434-634. In this paper, a 70 nt RNA sequence, corresponding to nt 480-550, was identified that binds TS protein with an affinity similar to that of full-length TS mRNA and TS434-634 RNA. In vitro translation studies confirmed that this sequence is critical for the translational autoregulatory effects of TS. To document in vivo biological significance, TS sequences contained within this region were cloned onto the 5' end of a luciferase reporter plasmid and transient transfection experiments were performed using H630 human colon cancer cells. In cells transfected with p644/TS434-634 or p644/TS480-550, luciferase activity was decreased 2.5-fold when compared to cells transfected with p644 plasmid alone. Luciferase mRNA levels were identical for each of these conditions as determined by RNase protection and RT-PCR analysis. Immunoprecipitation of TS ribonucleoprotein complexes revealed a direct interaction between TS protein and TS480-550 RNA in transfected H630 cells. Treatment with 5-fluorouridine resulted in a nearly 2-fold increase in luciferase activity only in cells transfected with p644/TS434-634 and p644/TS480 550. This study identifies a 70 nt TS response element in the protein coding region of TS mRNA with in vitro and in vivo translational regulatory activity. PMID- 10684934 TI - Different roles for abf1p and a T-rich promoter element in nucleosome organization of the yeast RPS28A gene. AB - In vivo mutational analysis of the yeast RPS28A ribosomal protein (rp-)gene promoter demonstrated that both the Abf1p binding site and the adjacent T-rich element are essential for efficient transcription. In vivo Mnase and DNaseI digestion showed that the RPS28A promoter contains a 50-60 bp long nucleosome free region directly downstream from the Abf1p binding site, followed by an ordered array of nucleosomes. Mutating either the Abf1p binding site or the T rich element has dramatic, but different, effects on the local chromatin structure. Failure to bind Abf1p appears to cause nucleosome positioning to become disorganized as concluded from the complete disappearance of Mnase hypersensitive sites. On the other hand, mutation of the T-rich element causes the downstream nucleosomal array to shift by approximately 50 bp towards the Abf1p site, resulting in loss of the nucleosome-free region downstream of Abf1p. We conclude that Abf1p is a strong organizer of local chromatin structure that appears to act as a nucleosomal boundary factor requiring the downstream T-rich element to create a nucleosome-free region. PMID- 10684935 TI - Genome sequences of Chlamydia trachomatis MoPn and Chlamydia pneumoniae AR39. AB - The genome sequences of Chlamydia trachomatis mouse pneumonitis (MoPn) strain Nigg (1 069 412 nt) and Chlamydia pneumoniae strain AR39 (1 229 853 nt) were determined using a random shotgun strategy. The MoPn genome exhibited a general conservation of gene order and content with the previously sequenced C.trachomatis serovar D. Differences between C.trachomatis strains were focused on an approximately 50 kb 'plasticity zone' near the termination origins. In this region MoPn contained three copies of a novel gene encoding a >3000 amino acid toxin homologous to a predicted toxin from Escherichia coli O157:H7 but had apparently lost the tryptophan biosyntheis genes found in serovar D in this region. The C. pneumoniae AR39 chromosome was >99.9% identical to the previously sequenced C.pneumoniae CWL029 genome, however, comparative analysis identified an invertible DNA segment upstream of the uridine kinase gene which was in different orientations in the two genomes. AR39 also contained a novel 4524 nt circular single-stranded (ss)DNA bacteriophage, the first time a virus has been reported infecting C. pneumoniae. Although the chlamydial genomes were highly conserved, there were intriguing differences in key nucleotide salvage pathways: C.pneumoniae has a uridine kinase gene for dUTP production, MoPn has a uracil phosphororibosyl transferase, while C.trachomatis serovar D contains neither gene. Chromosomal comparison revealed that there had been multiple large inversion events since the species divergence of C.trachomatis and C.pneumoniae, apparently oriented around the axis of the origin of replication and the termination region. The striking synteny of the Chlamydia genomes and prevalence of tandemly duplicated genes are evidence of minimal chromosome rearrangement and foreign gene uptake, presumably owing to the ecological isolation of the obligate intracellular parasites. In the absence of genetic analysis, comparative genomics will continue to provide insight into the virulence mechanisms of these important human pathogens. PMID- 10684936 TI - A new double-stranded RNA-binding protein that interacts with PKR. AB - We have identified a 74 kDa double-stranded (ds)RNA-binding protein that shares extensive homology with the mouse spermatid perinuclear RNA-binding (Spnr) protein. p74 contains two dsRNA-binding motifs (dsRBMs) that are essential for preferential binding to dsRNA. Previously, dsRNA-binding proteins were shown to undergo homo- and heterodimerization, raising the possibility that regulation of activity could be controlled by interactions between different family members. Homodimerization is required to activate the dsRNA-dependent protein kinase PKR, whereas hetero-dimerization between PKR and other dsRNA-binding proteins can inhibit kinase activity. We have found that p74 also interacts with PKR, both the wild-type enzyme and a catalytically defective mutant (K296R). While co expression of p74 and wild-type PKR in the yeast Saccharomyces cerevisiae did not alter PKR activity, co-expression of p74 and the catalytically defective K296R mutant surprisingly resulted in abnormal morphology and cell death in transformants that maintained a high level of p74 expression. These transformants could be rescued by overexpression of the alpha-subunit of wild-type eukaryotic translation initiation factor 2 (eIF2alpha), one of the known substrates for PKR. We hypothesize that competing heterodimers between p74-K296R PKR and eIF2alpha K296R PKR may control cell growth such that stabilization of the p74-K296R PKR heterodimer induces abnormal morphology and cell death. PMID- 10684937 TI - Determination of L1 retrotransposition kinetics in cultured cells. AB - L1 retrotransposons are autonomous retroelements that are active in the human and mouse genomes. Previously, we developed a cultured cell assay that uses a neomycin phosphotransferase ( neo ) retrotransposition cassette to determine relative retrotransposition frequencies among various L1 elements. Here, we describe a new retrotransposition assay that uses an enhanced green fluorescent protein (EGFP) retrotransposition cassette to determine retrotransposition kinetics in cultured cells. We show that retrotransposition is not detected in cultured cells during the first 48 h post-transfection, but then proceeds at a continuous high rate for at least 16 days. We also determine the relative retrotransposition rates of two similar human L1 retrotransposons, L1(RP)and L1.3. L1(RP)retrotransposed in the EGFP assay at a rate of approximately 0.5% of transfected cells/day, approximately 3-fold higher than the rate measured for L1.3. We conclude that the new assay detects near real time retrotransposition in a single cell and is sufficiently sensitive to differentiate retrotransposition rates among similar L1 elements. The EGFP assay exhibits improved speed and accuracy compared to the previous assay when used to determine relative retrotransposition frequencies. Furthermore, the EGFP cassette has an expanded range of experimental applications. PMID- 10684938 TI - Requirement for PCNA and RPA in interstrand crosslink-induced DNA synthesis. AB - Proliferating nuclear cell antigen (PCNA) and replication protein A (RPA) have proven to be essential elements in many aspects of DNA metabolism including replication, repair and recombination. We have developed an in vitro assay in which the presence of an interstrand crosslink stimulates the incorporation of radiolabeled nucleotides into both damaged and undamaged plasmid DNAs. Using this assay we have investigated the roles of PCNA and RPA in crosslink-induced DNA synthesis. p21, a potent inhibitor of PCNA, was found to strongly inhibit crosslink-induced incorporation. Addition of exogenous PCNA partially restored the resynthesis activity. Likewise, neutralization of RPA by monoclonal antibodies also inhibited incorporation, but the effect was somewhat more pronounced on the undamaged plasmid than the damaged plasmid. Addition of excess RPA also partially reversed antibody inhibition. These results indicate that both PCNA and RPA are required for efficient in vitro DNA resynthesis induced by interstrand crosslinks. PMID- 10684939 TI - Functional alpha-fragment of beta-galactosidase can be expressed from the mobile group I intron PpLSU3 embedded in yeast pre-ribosomal RNA derived from the chromosomal rDNA locus. AB - PpLSU3, a mobile group I intron found in the ribo-somal RNA genes of Physarum polycephalum, encodes the I-PpoI homing endonuclease. This enzyme represents one of the rare cases in nature where a protein is expressed from an RNA polymerase I transcript. Our previous results showed that the full length intron, but not a further processed species, is the messenger for I-PpoI, implying a role of the untranslated region (UTR) in gene expression. To study the function of the 3'-UTR in expression of the endonuclease and in splicing of the intron, we replaced the I-PpoI gene in PpLSU3 with the gene for the alpha-fragment of Escherichia coli beta-galactosidase, and then integrated this chimeric intron into all the chromosomal rDNA repeats of yeast. The resulting cells synthesized functional alpha-fragment, as evidenced by a complementation assay analogous to that used in E.coli. The beta-galactosidase activity thus provides an unusual and potentially valuable readout for Pol I transcription from chromosomal rDNA. This is the first example in which a eucaryotic homing endonuclease gene has been successfully replaced by a heterologous gene. Using deletion mutagenesis and a novel randomization approach with the alpha-fragment as a reporter, we found that a small segment of the 3'-UTR dramatically influences both splicing and protein expression. PMID- 10684940 TI - The initiator element of the Drosophila beta2 tubulin gene core promoter contributes to gene expression in vivo but is not required for male germ-cell specific expression. AB - The tissue-specific expression of the Drosophila beta 2 tubulin gene ( B2t ) is accomplished by the action of a 14-bp activator element (beta2UE1) in combination with certain regulatory elements of the TATA-less, Inr-containing B2t core promoter. We performed an in vivo analysis of the Inr element function in the B2t core promoter using a transgenic approach. Our experiments demonstrate that the Inr element acts as a functional cis -regulatory element in vivo and quantitatively regulates tissue-specific reporter expression in transgenic animals. However, our mutational analysis of the Inr element demonstrates no essential role of the Inr in mediating tissue specificity of the B2t promoter. In addition, a downstream element seems to affect promoter activity in combination with the Inr. In summary, our data show for the first time the functionality of the Inr element in an in vivo background situation in Drosophila. PMID- 10684941 TI - Ligation reaction specificities of an NAD(+)-dependent DNA ligase from the hyperthermophile Aquifex aeolicus. AB - An NAD(+)-dependent DNA ligase from the hyperthermophilic bacterium Aquifex aeolicus was cloned, expressed in Escherichia coli and purified to homogeneity. The enzyme is most active in slightly alkaline pH conditions with either Mg(2+)or Mn(2+)as the metal cofactor. Ca(2+)and Ni(2+)mainly support formation of DNA adenylate intermediates. The catalytic cycle is characterized by a low k (cat)value of 2 min(-1)with concomitant accumulation of the DNA - adenylate intermediate when Mg(2+)is used as the metal cofactor. The ligation rates of matched substrates vary by up to 4-fold, but exhibit a general trend of T/A < or = G/C < C/G < A/T on both the 3'- and 5'-side of the nick. Consistent with previous studies on Thermus ligases, this Aquifex ligase exhibits greater discrimination against a mismatched base pair on the 3'-side of the nick junction. The requirement of 3' complementarity for a ligation reaction is reaffirmed by results from 1 nt insertions on either the 3'- or 5'-side of the nick. Furthermore, most of the unligatable 3' mismatched base pairs prohibit formation of the DNA-adenylate intermediate, indicating that the substrate adenylation step is also a control point for ligation fidelity. Unlike previously studied ATP ligases, gapped substrates cannot be ligated and intermediate accumulation is minimal, suggesting that complete elimination of base pair complementarity on one side of the nick affects substrate adenylation on the 5' side of the nick junction. Relationships among metal cofactors, ligation products and intermediate, and ligation fidelity are discussed. PMID- 10684942 TI - Pre-selection of integration sites imparts repeatable transgene expression. AB - Variable gene expression amongst transgenic lines occurs due to copy number and to random associations of incoming DNA with chromosomal elements at the site of integration. Here we describe a method of identifying sites permissive for transgene expression and their use for efficient introduction of single copy transgenes by homologous recombination. ES clones were selected in HAT medium for expression of a randomly integrated HPRT marker lying 5' to an Oct4/ lacZ transgene. 794 clones were assessed in vitro for appropriate down-regulation of lacZ following differentiation. Two clones were chosen for further analysis which displayed appropriate and inappropriate gene regulation (clones 710 and 91, respectively). Three developmental promoters (thyroglobulin, Hox2.6 and Myf5) were then sequentially introduced into the original insertion sites in each clone (710 and 91) by homologous recombination, to drive expression of lacZ. Transgenic embryos were assessed for their ability to direct lacZ expression to tissues in which the respective promoter sequences are normally active. The site which appropriately down-regulated lacZ in vitro (710) also showed appropriate in vivo regulation of lacZ from the three developmental promoters. Site 91, however, directed an additional pattern of ectopic expression, which was common to all four promoters. Pre-selection of genomic sites for the introduction of transgenes by gene targeting improves the repeatability of transgene expression and provides an efficient means of single copy transgene introduction by homologous recombination. PMID- 10684943 TI - A developmentally regulated deletion element with long terminal repeats has cis acting sequences in the flanking DNA. AB - Approximately 6000 specific DNA deletion events occur during development of the somatic macro-nucleus of the ciliate Tetrahymena. The eliminated Tlr1 element is 13 kb or more in length and has an 825 bp inverted repeat near the rearrangement junctions. A functional analysis of the cis -acting sequences required for Tlr1 rearrangement was performed. A construct consisting of the entire inverted repeat and several hundred base pairs of flanking DNA on each side was rearranged accurately in vivo and displayed junctional variability similar to the chromosomal Tlr1 rearrangement. Thus, 11 kb or more of internal element DNA is not required in cis for DNA rearrangement. A second construct with only 51 bp of Tetra-hymena DNA flanking the right junction underwent aberrant rearrangement. Thus, a signal for determination of the Tlr1 junction is located in the flanking DNA, 51 bp or more from the right junction. Within the Tlr1 inverted repeat are 19 bp tandem repeats. A construct with the 19mer repeat region deleted from the right half of the inverted repeat utilized normal rearrangement junctions. Thus, despite its transposon-like structure, Tlr1 is similar to other DNA rearrangements in Tetrahymena in possessing cis -acting sequences outside the deleted DNA. PMID- 10684944 TI - Cloning and characterisation of the Sry-related transcription factor gene Sox8. AB - SOX proteins form a large family of transcription factors related by a DNA binding domain known as the HMG box. Some 30 Sox genes have been identified in mammals and orthologues have been found in a wide range of other metazoans. Sox genes are highly conserved and are known to play important roles in embryonic development, including roles in gonadal, central nervous system, neural crest and skeletal development. Several SOX genes have been implicated in human congenital diseases. We report here the isolation of Sox8 and its characterisation in mice and humans. This gene has a remarkably similar primary structure and genomic organisation to the campomelic dysplasia gene SOX9 and the Waardenburg-Shah syndrome gene SOX10. SOX8 protein is able to bind to canonical SOX target DNA sequences and activate transcription in vitro through two separate trans activation regions. Further, Sox8 is expressed in the central nervous system, limbs, kidneys, gonads and craniofacial structures during mouse embryo development. Sox8 maps to the t complex on mouse chromosome 17 and to human chromosome 16p13.3, a region associated with the microphthalmia-cataract syndrome CATM and the alpha-thalassemia/mental retardation syndrome ATR-16. PMID- 10684945 TI - Analysis of the yeast transcriptome with structural and functional categories: characterizing highly expressed proteins. AB - We analyzed 10 genome expression data sets by large-scale cross-referencing against broad structural and functional categories. The data sets, generated by different techniques (e.g. SAGE and gene chips), provide various representations of the yeast transcriptome (the set of all yeast genes, weighted by transcript abundance). Our analysis enabled us to determine features more prevalent in the transcriptome than the genome: i.e. those that are common to highly expressed proteins. Starting with simplest categories, we find that, relative to the genome, the transcriptome is enriched in Ala and Gly and depleted in Asn and very long proteins. We find, furthermore, that protein length and maximum expression level have a roughly inverse relationship. To relate expression level and protein structure, we assigned transmembrane helices and known folds (using PSI-blast) to each protein in the genome; this allowed us to determine that the transcriptome is enriched in mixed alpha-beta structures and depleted in membrane proteins relative to the genome. In particular, some enzymatic folds, such as the TIM barrel and the G3P dehydrogenase fold, are much more prevalent in the transcriptome than the genome, whereas others, such as the protein-kinase and leucine-zipper folds, are depleted. The TIM barrel, in fact, is overwhelmingly the 'top fold' in the transcriptome, while it only ranks fifth in the genome. The most highly enriched functional categories in the transcriptome (based on the MIPS system) are energy production and protein synthesis, while categories such as transcription, transport and signaling are depleted. Furthermore, for a given functional category, transcriptome enrichment varies quite substantially between the different expression data sets, with a variation an order of magnitude larger than for the other categories cross-referenced (e.g. amino acids). One can readily see how the enrichment and depletion of the various functional categories relates directly to that of particular folds. PMID- 10684946 TI - Species-specific regulation of alternative splicing in the C-terminal region of the p53 tumor suppressor gene. AB - Alternative splicing occurs in the C-terminal region of the p53 tumor suppressor gene between two alternative 3' splice sites in intron 10. This alternative splicing event has been detected in murine cells, but not in rat or human tissues. In this paper, we have characterized the pattern of p53 alternative splicing in cell lines from five different species. Our results confirm that p53 alternative splicing is species-specific, being detected only in cell lines of rodent origin. Using transient transfection assays, we have established that the rat p53 gene undergoes efficient alternative splicing in both mouse and rat cell lines, thus demonstrating that it has all the necessary cis -acting sequences to be alternatively spliced. In contrast, we were unable to detect any usage of the human alternative 3' splice site under the same experimental conditions. Thus, the low levels or absence of alternatively spliced p53 mRNA in rat and human cell lines seems to be the result of different mechanisms. Our results support the hypothesis that there are species-specific mechanisms implicated in the regulation of p53 activity. PMID- 10684947 TI - A rapid genetic screening system for identifying gene-specific suppression constructs for use in human cells. AB - We describe a rapid cell-based genetic screen using fission yeast for identifying efficient gene suppression constructs (GSCs) from large libraries (10(5)) for any target sequence for use in human cells. In this system, target sequences are fused to the 5' end of the lacZ reporter gene and expressed in yeast. Random fragment expression libraries derived from the target sequence are screened in the fusion gene-expressing strain using the lacZ gene-encoded colony color phenotype. We demonstrate the utility of this screening assay by identifying a range of different GSCs for the fission yeast ura4 gene and human c-myc and Chk1 sequences, including rare efficient suppressors. GSCs specific for c-myc were shown to regulate expression of both a c-myc-lacZ fusion gene and the endogenous c-myc gene in human cells. PMID- 10684949 TI - Editorial PMID- 10684948 TI - Endogenous oxidative DNA base modifications analysed with repair enzymes and GC/MS technique. AB - GC/MS technique was used to identify endogenous levels of oxidatively modified DNA bases. To avoid possible artefact formation we used Fpg and Endo III endonucleases instead of acid hydrolysis to liberate the base products from unmodified DNA samples. Several different DNA preparations were used: (i) commercial calf thymus DNA, (ii) DNA isolated from rat liver, (iii) DNA isolated from human lymphocytes and (iv) nuclei isolated from rat liver. In all DNA samples used in our assays the most efficiently removed bases by Fpg protein are FapyG and FapyA although 8-oxoG was also detected in all preparations. The amount of 8-oxoG in human lymphocytes and in rat liver DNA was 3 and 2 per 10(7)bases, respectively. It is reasonable to postulate that the presented method is one of the techniques which should be used to reveal the enigma of endogenous, oxidative DNA damage. PMID- 10684950 TI - Stress, burnout and locus of control in German nurses. AB - The aim of this study was to evaluate the effects of locus of control and work related stress on burnout in hospital staff nurses. A convenience sample of 361 staff nurses from nine units in five German hospitals were surveyed using the Maslach Burnout Inventory, the Locus of Control Questionnaire and a Work-Related Stress Inventory. Causal modeling was used to explore the moderating effect of locus of control on burnout. Results support the hypothesized model and suggest that greater work-related stress and burnout would be associated with poorer locus of control in nurses. The findings supported the notion that perceived degree of control is instrumental in enabling nurses to cope with stress and burnout. PMID- 10684951 TI - Frequency of intravenous medication administration to hospitalised patients: secondary data-analysis of the Belgian nursing minimum data set. AB - The purpose of this study was to investigate the frequency of intravenous medication administration with Belgian hospitalised patients. Factors, which might influence this frequency of administration, were also studied. Research questions were investigated by secondary data-analysis of the Belgian Nursing Minimum Data Set. The randomised sample consisted of 1,035,681 observations on 421,530 patients. Results of this study demonstrate that one out of three (34%) hospitalised patients received intravenous medication. Medical diagnoses, for which most intravenous medications were administered, were oncological diseases: myeloid (77.9%) and lymphoid (69.4%) leukaemia. Elderly (6.7%) and female (31.2%) patients received significantly less intravenous medication than respectively young (32.9%) (chi(2) = 98411, df = 1, p<0.001) and male (38%) (chi(2) = 2033, df = 1, p<0.001) patients. Patients with intravenous medication administration were labour intensive for nursing staff. PMID- 10684952 TI - Living a restricted life with Ehlers-Danlos syndrome (EDS). AB - Ehlers-Danlos syndrome (EDS) is an inherited connective tissue disorder, primarily affecting the skin, ligaments, joints and blood vessels. The symptoms can vary from undiagnosed cases with mild symptoms to more severe forms. A qualitative study was conducted with the purpose of exploring how individuals with different symptoms of EDS describe their symptoms and perceive their daily life. Eleven interviewees were recruited via a support group in Sweden. The main strategies of the grounded theory method were used in collection and analysis of data. The identified main theme, "Living a restricted life", seemed to explain the way in which fears, pain, stigmatisation and experiences of non-affirmation in health-care limited the possibility of self-actualisation in daily living and social life. This study provides a conceptual framework for future research and an understanding of the type of professional support individuals with EDS require. PMID- 10684953 TI - Exercise participation decisions of Jordanian myocardial infarction patients: application of the decisional conflict theory. AB - This study examined the utility of Janis and Mann's [Janis, I. L. & Mann, L. (1977). Decision making: a psychological analysis conflict, choice and commitment. New York, NY: Free press.] decisional conflict theory (DCT) in predicting decisions to exercise following hospitalization for a myocardial infarction (MI). A structured interview of operationalized DCT variables was conducted with 88 MI patients attending cardiology clinics in two Jordanian military hospitals. Stress was viewed as an indicator of exercise benefits barriers decisional conflict. The findings revealed that the benefits-barriers interaction significantly predicted stress for the 48 patients who did not exercise. Rehospitalization for cardiac events was a significant predictor of stress for the 40 exercisers. Preinfarction exercise behavior, stress and gender were significant predictors of MI patients' decisions to exercise. Results of this study indicated that decisional conflict and stress were characteristics of MI patients who delayed or avoided exercising. PMID- 10684954 TI - Measurement of fatigue in chronic obstructive pulmonary disease and in asthma. AB - Despite recognition of a high prevalence of fatigue in individuals with chronic airflow obstruction conditions, including chronic obstructive pulmonary disease and asthma, and its importance from a quality of life perspective, no research was found in which fatigue was measured directly in these populations. This may be due to a seeming lack of appropriate instruments for measuring fatigue in these populations. The purpose of this study, therefore, was to pretest an instrument, the Piper Fatigue Scale, which was developed to measure chronic fatigue in clinical populations. The outpatient sample consisted of 17 persons with chronic obstructive pulmonary disease and 19 with asthma. Findings revealed that the visual analogue scale version of the Piper Fatigue Scale may not be appropriate for measuring fatigue in these populations. Instruments with validity and reliability for fatigue in chronic obstructive pulmonary disease and asthma need to be developed. PMID- 10684955 TI - Problem-solving skills of senior student nurses: an exploratory study using simulation. AB - The Stages Model of problem-solving as evidenced in the use of the nursing process is the key vehicle for the operationalisation of problem-solving in current nursing practice. An expanded variant of the model formed the theoretical framework for this study which aimed to explore and compare the problem-solving skills of senior student nurses (n=253) from three pre-registration nurse education programmes (RGN, diploma RN, integrated degree). Students' care planning skills were explored using a video-tape simulation exercise and data were subjected to statistical testing. Findings indicated a large range in the global care plan scores and while performance was similar in a number of areas independent of programme type, certain key differences also emerged. The findings are discussed in the wider context of professional education and practice and the potential for further development of problem-solving skills in pre-registration nurse education is explored. PMID- 10684956 TI - The art and science of predicting patient readiness for weaning from mechanical ventilation. AB - Weaning from mechanical ventilation is attempted when the patient's underlying condition has resolved and when the patient is able to maintain cardiovascular and respiratory stability within normal parameters. From a medical perspective, when to wean is based on patient readiness determined by objective, physiological criteria. Psychological readiness is equally important, yet criteria determining psychological readiness is generally omitted from the list of ready to wean parameters. Reasons for this may be that psychological readiness is difficult to measure and is based upon subjective opinions. Nursing research exploring critical care nurses' and patients' experiences of weaning has extended knowledge concerning patients' psychological readiness to wean. From examination of this research, three important criteria emerge. It is recommended that the addition of these criteria to the list of physiological criteria will not only achieve a holistic assessment of patients' readiness to wean, but will also acknowledge the important and complementary role of the nurse in the weaning process. PMID- 10684957 TI - The effectiveness of intermediate care in a nursing-led in-patient unit. AB - In order to assess the potential for a nursing-led in-patient unit (NLIU) to substitute for a period of care in the acute hospital environment and promote recovery before discharge, a randomised controlled trial was conducted. The setting was an acute inner London hospital trust, part of the UK's national health service. Of patients referred to a NLIU from acute wards, 80 were randomly assigned to usual care (remain in normal hospital system) and 97 to the NLIU (nursing-led care with no routine medical involvement). Patients were identified as medically stable but in need of additional nursing intervention by referring medical staff prior to full nursing assessment of suitability. Outcomes compared included functional dependence (Barthel Index), discharge destination and length of hospital stay. Inputs from nursing, paramedical and medical staff were measured. There was no significant difference in functional independence at discharge (p0.05). Patients undergoing usual care stayed in hospital for less time (mean difference 18 days, p<0.01) but the same number of patients were in hospital 90 days after recruitment (23% NLIU, 24% usual care p0.05) due to re admissions. The model of care implemented differed considerably from that described in the literature with the NLIU having significantly fewer qualified nurses (RNs). Although the anticipated benefits of the NLIU were not demonstrated, the study does not conclude that the model should be rejected. Factors driving length of stay need to be further investigated, as does the possibility of post-discharge benefits. The NLIU does offer some potential to substitute for acute care but also appears to substitute for a period of primary care. PMID- 10684958 TI - Effects of hospital restructuring on full time and part time nursing staff in Ontario. AB - This study examined the effects of hospital restructuring and downsizing on full time and part-time nursing staff. Data were collected from 1362 nursing staff, a 35% response rate, using anonymous questionnaires. Measures included personal and situational characteristics, hospital restructuring and downsizing variables, work outcomes and psychological well-being indicators, and work-family experiences. Although full and part-time nurses were significantly different on most personal and demographic characteristics, both groups experienced and described hospital restructuring and downsizing similarly. Full-time nurses reported greater emotional exhaustion and poorer health and indicated greater absenteeism and lower intention to quit. PMID- 10684959 TI - Why nurses smoke: a review of the literature. AB - The smoking behaviour of nurses has been widely debated in the context of their professional role and responsibilities. There has been much speculation about why nurses smoke and possible explanations include a stressful nursing environment, peer pressure and socio economic status and education. This paper provides an overview of the literature which offers insights into the reasons why nurses smoke and compares the findings from this literature with those studies examining the smoking behaviour of women in general and young women in particular. This review reveals that many students take up smoking before commencing their training and the factors which influence nurses smoking are similar to those that influence similar groups of females in the general population. PMID- 10684960 TI - Requirement of MEF2D in the induced differentiation of HL60 promyeloid cells. AB - The regulatory role of MEF2 (myocyte enhancer binding factor 2) proteins in nonmuscle tissues has not been well characterized. We examined the expression of MEF2 family members, namely, MEF2A, -B, -C, and -D, in the differentiation of HL60 promyeloid cells and observed the remarkable increase in the expressions of MEF2A and MEF2D proteins during the differentiation process into monocytes. To examine the role of MEF2, we expressed a dominant-negative form of MEF2D, without its transactivation domain, in HL60 cells. When the HL60 cell line expressing the mutant MEF2D was induced to differentiate by VitD(3) treatment, cell surface expression of CD14 and the ability to reduce NBT, which are important characteristics of differentiated monocytes, were significantly decreased compared with control HL60 cells. These results show that MEF2D is required in the differentiation process along the monocyte/macrophage lineage, PMID- 10684961 TI - Innate antibody catalysis. AB - Catalysis by antibodies is often assumed to require immunization with artificial haptens, which are proposed to stimulate adaptive immune processes and enable the development of catalytic sites with the ability to bind the transition state. Contrary to this assumption, we describe here a serine protease-like catalytic triad in an antibody light chain raised by immunization with vasoactive intestinal peptide (VIP), the structure and function of which is inherited via a germline V(L) gene. The serine protease mechanism was evident from loss of the catalytic activity by site directed mutagenesis at a framework region residue Asp1 (present study) and at two complementarity determining region residues Ser27a and His 93 (Gao, Q-S., Sun, M., Rees, A., Paul, S., 1995. Site-directed mutagenesis of proteolytic antibody light chain. J. Mol. Biol. 253, 658-664). All three catalytic residues (Ser27a, His93, Asp1) are also present in the germline counterpart of the mature V(L) gene, but the mature and germline sequences differ by four amino acids remote from the catalytic site. Reversion mutations were introduced at these amino acids in the mature light chain (His27 d:Asp, Thr28e:Ser, Ile34:Asn, Gln96:Trp; Kabat numbering, germline encoded residues shown second), generating the germline configuration of the protein. The germline light chain expressed peptidase activity, determined by assaying the cleavage of VIP and a synthetic protease substrate, Pro-Phe-Arg-Methylcoumarinamide. Differences between the kinetic constants for the mature and germline light chains were marginal. Diisopropylfluorophosphate, a serine protease inhibitor, blocked the peptidase activity of the germline light chain, suggesting the presence of the catalytic triad in a functional state. Like the mature light chain, the germline protein preferentially cleaves peptide bonds on the C terminal side of basic residues. We conclude that the catalytic activity of certain antibodies is an innate function, originating over the course of phylogenetic evolution of the V(L) genes, as opposed to somatic processes. PMID- 10684962 TI - Characterization of novel FcepsilonRII/CD23 isoforms lacking the transmembrane (TM) segment in human cell lines. AB - Human FcepsilonRII/CD23 is an approximately 45 kDa type II transmembrane glycoprotein belonging to the C-type animal-lectin family, and has two isoforms (a and b) that only differ in their intracytoplasmic tails. We previously found that in several human and mouse cell lines there were two additional CD23 transcripts (a' and b') lacking the exon 3 that encodes the entire transmembrane segment and a part of cytoplasmic tails. In this study, we analyzed the putative CD23a' and CD23b' products at protein levels and characterized with rabbit polyclonal antibodies against novel amino-acid sequences of the putative CD23a' and CD23b' molecules (anti-CD23a' Ab, anti-CD23b' Ab). Western blots in COS cells transfected with CD23a' or CD23b' cDNA as well as in vitro translation assays showed that the a' and b' CD23 transcripts were translated to about 40 kDa molecules. These 40 kDa molecules were also recognized by a polyclonal antibody against 25 kDa soluble fragment of human CD23. We also found that human cells having mRNAs for CD23a' and CD23b' expressed protein products recognized specifically by anti-CD23a' or anti-CD23b' Ab, respectively. In addition, the CD23a' and CD23b' molecules in transfected COS cells were resistant to Endo H(f) and PNGase F, although these truncated forms as well as the membrane-associated forms had an asparagine residue responsible for the N-linked glycosylation. Taken together, our results show that the a' and b' CD23 transcripts are expressed and translated in human lymphoid cells and that their translated products are retained in the cytoplasm where they might play an unique regulatory role in the expression of the full-length CD23 on the cell surface. PMID- 10684963 TI - Selection of phage-displayed anti-guinea pig C5 or C5a antibodies and their application in xenotransplantation. AB - Xenogeneic liver transplantation in the discordant guinea pig (gp) to rat model results in hyperacute rejection within a few minutes, which is due to activation of the complement system. Currently no antibodies against gp complement factors are available, which allow activation of the gp complement system in serum or complement deposition in tissue to be detected. To close this gap, we started developing single chain Fvs (scFvs) against gpC5 and gpC5a. We generated a combinatorial library of scFv antibodies comprising the variable heavy and light chain repertoire from mice immunized with gpC5. Out of this library we selected several antibodies against gpC5 and C5a after four and six rounds of biopanning, respectively. Selected gpC5-specific scFvs were purified by metal affinity chromatography followed by size exclusion chromatography or by affinity chromatography using Protein L. Purified scFvs were able to inhibit gp complement system in a hemolytic assay and to detect gpC5 deposition in tissue. A surface plasmon resonance based assay on BIAcore was established, with which the C5 concentration in gp serum was determined to 240 microg/ml. As at least 0.04% of the normal gpC5 concentration can be detected, the test provides a powerful tool to investigate the development and the consequence of a hybrid complement system after liver xenotransplantation from gp to rat. PMID- 10684964 TI - Inhibition of M-tropic HIV-1 infection by the fd phage-gene 3 protein with MIP 1alpha-binding activity. AB - CCR5 is a chemokine receptor with seven transmembrane-domains. It is expressed on T cells and macrophages and functions as the principal co-receptor for macrophage (M)-tropic strains of HIV-1. The anti-CCR5 monoclonal antibody (mAb) 2D7 inhibits the binding and chemotaxis of the three natural beta-chemokine ligands of CCR5, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES, to CCR5(+) cells. The mAb also efficiently blocks the infectivity of several M-tropic and dual-tropic HIV-1 strains in vitro. In this study, we attempted to determine the peptide motif recognized with the 2D7 mAb. We isolated phage clones by panning a phage display library using 2D7 and identified three peptide motifs. One of these phage clones (M23) showed a marked inhibitory activity on HIV-1 infection. The unique sequence of 15 amino acids with an internal disulfide bond was inserted in the g3p of the M23 phage clone (M23-g3p). The M23-g3p was purified by fast performance liquid chromatography (FPLC). We show here that (1) M23-g3p was specifically recognized with anti-CCR5 mAb; (2) M23-g3p showed inhibitory activity on the infectivity of M-tropic but not T-tropic HIV-1 strains; (3) M23 g3p bound to MIP-1alpha, MIP-1beta, and RANTES but not MCP-1. These results suggested that the M23-g3p might mimic the CCR5-binding domain shared by beta chemokines, MIP-1alpha, MIP-1beta, and RANTES as well as the HIV-1 infection. PMID- 10684965 TI - Molecular cloning of the brushtail possum (Trichosurus vulpecula) immunglobulin E heavy chain constant region. AB - The immunobiology of marsupial IgE is poorly understood. As a first step towards the development of immunological reagents for marsupials and to obtain a further understanding of immunoglobulin evolution, a brushtail possum (Trichosurus vulpecula) mesenteric lymph node cDNA library was screened for the heavy chain constant region of IgE (Cepsilon), using a partial Cepsilon probe from the American marsupial, Monodelphis domestica. The cDNA sequence for T. vulpecula Cepsilon was determined and found to be most similar to the M. domestica Cepsilon sequence [(76%) at the amino acid level]. T. vulpecula Cepsilon has amino acid sequence similarities ranging from 43-52% with various eutherian Cepsilon sequences. The secondary structure of T. vulpecula Cepsilon, based on loops formed by internal disulfide bonds, more closely resembles rodent Cepsilon than the American marsupial sequence. PMID- 10684966 TI - V(D)J recombination catalyzed by mutant RAG proteins lacking consensus DNA-PK phosphorylation sites. AB - The process of antigen receptor gene rearrangement, V(D)J recombination, involves DNA cleavage by the RAG-1 and RAG-2 proteins. Cleavage generates covalently sealed (hairpin) DNA ends, termed coding ends, which must be opened by an endonuclease prior to joining. Resolution of these hairpin ends requires the activity of the DNA-dependent protein kinase (DNA-PK), a protein kinase whose specific role is yet undetermined. It has been suggested that phosphorylation of one or both RAG proteins by DNA-PK is required to activate or recruit the hairpin opening nuclease. Furthermore, very recent work has shown that RAG proteins themselves can open hairpins. These data raise the possibility that DNA-PK mediated phosphorylation of the RAG proteins could regulate the hairpin opening reaction. To test this hypothesis, we constructed mutant versions of RAG-1 and RAG-2 in which all four DNA-PK consensus phosphorylation sites were removed by site-directed mutagenesis. Our data provide conclusive evidence that phosphorylation of these conserved serine/threonine residues is not required for hairpin opening or joining of V(D)J recombination intermediates. PMID- 10684967 TI - A novel DNA element mediates transcription of Nkx2.1 by Sp1 and Sp3 in pulmonary epithelial cells. AB - NKX2.1 is a member of the NK2 family of homeodomain-containing transcription factors whose targeted disruption in mouse results in the absence of thyroid tissue and a severely abnormal lung phenotype. Little is known regarding the mechanisms that control tissue and temporal specificity of Nkx2.1 gene expression. The Nkx2.1 gene has been cloned from a number of species and it is composed of three exons and two introns. Two distinct DNA domains located 5' of exon I and within intron I have been found to exhibit promoter activity in lung and thyroid cells. In the current study we used deletional analysis of the 5' flanking region of exon I and identified a 300 bp TATA-less region that exhibits significant promoter activity in H441 cells. The DNA sequence of this region contains multiple palindromes, composed of G/C-rich elements. DNase I footprinting demonstrates that this promoter region interacts with nuclear factors present in H441 cells. In particular electrophoretic mobility shift assay using antibodies against the Sp family members show that both Sp1 and Sp3 as well as an as yet unknown H441-specific factor interact with the palindromic structure within this promoter region. Co-transfection studies show that this promoter region responds to Sp1 and Sp3 and mutations therein result in a significantly diminished response to these transcriptional factors. Therefore, we have identified a novel DNA structure on the Nkx2.1 gene which participates in transcription of this gene in pulmonary epithelial cells by Sp1 and Sp3 transcription factors. PMID- 10684968 TI - The intron-containing L3 ribosomal protein gene (RPL3): sequence analysis and identification of U43 and of two novel intronic small nucleolar RNAs. AB - Isolation and sequencing of bovine and human intron-containing L3 ribosomal protein genes are here reported. They exhibit very similar organisation, both comprising 10 exons and nine introns. A polymorphic locus, involving a 19-bp deletion, was found in intron 6 of the human gene. The frequency of the two alleles has been estimated in 200 haploid genomes. In bovine and human genes intron sequences are rather different, except for limited regions, located in corresponding positions, which show a surprisingly high degree of identity. All these regions contain conserved features defining the box C/D class of small nucleolar RNAs. Demonstration is given that U43 small nucleolar RNA is encoded within the first intron of both bovine and human genes. Single nucleotide sequences, encoding two novel species of small nucleolar RNAs (U82, U83a and U83b), are located in introns 3, 5 and 7. Their expression has been investigated and a possible role of these molecules in 2'-O-ribose methylation of rRNAs is discussed. PMID- 10684969 TI - Molecular characterization of ubiquitin genes from Aspergillus nidulans: mRNA expression on different stress and growth conditions. AB - We are interested in studying the ubiquitin (UBI) gene expression during different stress and growth conditions in the filamentous fungus Aspergillus nidulans. Here, we report the cloning of a cDNA clone that corresponds to a gene, ubi1, that encodes a carboxyl extension protein from A. nidulans. This cDNA corresponds to a gene that encodes a protein that showed high homology to other polyubiquitin and CEP-80 genes at the N- and C-terminus, respectively. We characterize the mRNA expression of the CEP and polyubiquitin genes during several growth and stress conditions. Expression of the ubi1 and ubi4 genes was correlated with cell growth in most of the carbon sources used, except maltose. Both ubi1 and ubi4 genes were induced upon heat-shock, although the levels of expression were raised quicker for ubi4 than for ubi1. The ubi1 and ubi4 genes displayed a very complex expression pattern in presence of drugs with a different mechanism of action suggesting that the regulatory processes controlling UBI gene expression discriminate between different stresses and can affect individually each UBI gene. The ubi1 gene was highly expressed in presence of hydrogen peroxide while the ubi4 mRNA level was not affected; several metals in our experimental conditions were not able to induce either ubi1 nor ubi4 genes. PMID- 10684970 TI - Expression and characterization of the human mitochondrial leucyl-tRNA synthetase. AB - A cDNA clone encoding the human mitochondrial leucyl-tRNA synthetase (mtLeuRS) has been identified from the EST databases. Analysis of the protein encoded by this cDNA indicates that the protein is 903 amino acids in length and contains a mitochondrial signal sequence that is predicted to encompass the first 21 amino acids. Sequence analysis shows that this protein contains the characteristic motifs of class I aminoacyl-tRNA synthetases and regions of high homology to other mitochondrial and bacterial LeuRS proteins. The mature form of this protein has been cloned and expressed in Escherichia coli. Gel filtration indicates that human mtLeuRS is active in a monomeric state, with an apparent molecular mass of 101 kDa. The human mtLeuRS is capable of aminoacylating E. coli tRNA(Leu). Its activity is inhibited at high levels of either monovalent or divalent cations. K(M) and k(cat) values for ATP:PP(i) exchange and for the aminoacylation reaction have been determined. PMID- 10684971 TI - Transforming growth factor beta inhibitory element in the rabbit matrix metalloproteinase-1 (collagenase-1) gene functions as a repressor of constitutive transcription. AB - Transforming growth factor beta (TGF-beta) is a potent modulator of the extracellular matrix, enhancing collagen synthesis and regulating expression of several genes that encode the matrix metalloproteinases (MMPs), enzymes that degrade the extracellular matrix. In this study, we explored the mechanisms whereby TGF-beta inhibits expression of the MMP-1 (collagenase 1) gene. We used transient transfection and gel mobility shift assays to characterize a TGF beta inhibitory element (TIE) at -249 bp in the rabbit MMP-1 promoter, which is also conserved at -246 bp in the human gene. This sequence shares homology to a previously identified TIE in the rat stromelysin-1 (MMP-3) promoter, where it is located at -709 bp. Mutational analyses and transient transfections indicate that MMP-1 TIE functions both as a constitutive repressor of MMP-1 gene expression and, in the presence of TGF-beta, as an antagonist of transcriptional induction by phorbol esters. c-Fos binds to the TIE in the rabbit MMP-1 promoter, along with other nuclear proteins, even in the absence of treatment with TGF-beta. However, the pattern of proteins binding to the TIE is altered in the presence of nuclear extracts from TGF-beta-treated cells, suggesting that TGF-beta leads to an alteration in protein/DNA interaction, with subsequent modulation of MMP-1 gene expression. We conclude that in the rabbit MMP-1 promoter, the TIE has dual functions as a repressor of basal transcription and as a mediator of the biologic effects of TGF-beta. Furthermore, these dual functions provide additional and subtle mechanisms for regulating MMP-1 gene expression under a variety of biological and pathological conditions. PMID- 10684972 TI - Heterologous expression of clostridial hydrogenase in the Cyanobacterium synechococcus PCC7942. AB - The Clostridium pasteurianum hydrogenase I has been expressed in the cyanobacterium Synechococcus PCC7942. The Shine-Dalgarno sequence of the structural gene encoding hydrogenase I from C. pasteurianum was changed to that of the cat (chloramphenicol acetyltransferase) gene. The hydrogenase gene was cloned downstream of a strong promoter, isolated from Synechococcus PCC7942, with the cat gene as a reporter gene. Expression of clostridial hydrogenase was confirmed by Western and Northern blot analyses in Synechococcus and Escherichia coli, whereas in vivo/in vitro measurements and activity staining of soluble proteins separated on non-denaturing polyacrylamide gels revealed functional expression of hydrogenase only in cyanobacterial cells. The changed Shine Dalgarno sequence appeared to be essential for the functional expression of clostridial hydrogenase in Synechococcus, but had no influence on the expression and activity of clostridial hydrogenase expressed in E. coli. PMID- 10684973 TI - Cloning and expression of the genes involved in the production of and immunity against the bacteriocin lacticin RM. AB - The production of lacticin RM, a novel bacteriocin produced by Lactococcus lactis subsp. lactis EZ26, is associated with the presence of a 6-kb plasmid, pHU1. The information necessary for lacticin RM production and immunity was localized to a 2.5-kb SalI-Eco47III fragment. Sequencing analysis of this fragment revealed the presence of six open reading frames (ORFs). Deletion and mutation analyses showed that orfX and orfY are not required for lacticin RM production or immunity, whereas the other ORFs (lacA, lacF, lacG and lacI) are necessary for the bacteriocin's production. Transcription analysis indicated that lacA, lacF and lacG are organized in an operon. lacA is probably the lacticin RM structural gene. It putatively encodes a 134-amino acid peptide, and it does not share homology with known bacteriocins. The deduced LacG protein is hydrophobic and consists of six potential trans-membrane helices. lacF encodes a conserved ATP binding domain homologous to ABC transporters known in bacteriocin immunity systems. LacF and LacG may form an active ABC transporter. Gene-disruption mutations indicated that both are required for immunity against lacticin RM. lacI encodes a small cationic protein, which is required for the production of and immunity to lacticin RM. Protection was obtained only when lacF, lacG and lacI were present together. PMID- 10684974 TI - Molecular characterization of the mouse Fas ligand promoter in airway epithelial cells. AB - Constitutively expressed Fas ligand in several distinct epithelial cell types appears to protect tissues by inducing apoptosis of Fas(+) immune cells during inflammatory reactions. To study the transcriptional regulation of Fas ligand gene in airway epithelial cells, a 618-bp 5'-flanking region of mouse Fas ligand gene was cloned, sequenced, and the transcriptional start site was determined by using 5'-RACE. Deletion analysis, gel mobility shift assays and site-directed mutagenesis indicated that a CCAAT box located -214 bp upstream from the transcription start site served as a major positive regulatory cis-element in an airway epithelial cell line. This element was not required for constitutive Fas ligand expression in Sertoli cells. Furthermore, the activity of the site did not involve the NF-Y protein complex or c/EBP protein family. UV-cross linking proteins to this element indicated that a approximately 23-kDa transcription factor bound to the Fas ligand promoter CCAAT box and, thus, likely plays an important role in the regulation of Fas ligand expression in airway epithelial cells. PMID- 10684975 TI - Sp1 mediates constitutive and transforming growth factor beta-inducible expression of urokinase type plasminogen activator receptor gene in human monocyte-like U937 cells. AB - Urokinase type plasminogen activator receptor (uPAR) is known to be involved in conversion of plasminogen into plasmin and its expression can be regulated by a variety of biological agents including transforming growth factor beta (TGF beta). In the present study, we cloned the promoter region of the human uPAR (huPAR) gene (-653 to +61) and investigated the transcription regulatory mechanism of the expression of the huPAR gene upon treatment with TGF-beta in human monocyte-like U937 cells. By deletion and point mutational analysis of the huPAR gene promoter, it was found that the sequence positioned at -70 is required for both constitutive and TGF-beta-inducible expression of the huPAR gene in U937 cells. Using electrophoretic mobility shift assay, we could observe that Sp1 formed a DNA-protein complex at the -70 sequence. In addition, antisense oligonucleotide against human Sp1 blocked both constitutive and TGF-beta inducible expression of the luciferase reporter gene driven by the huPAR gene promoter in U937 cells. These results led us to conclude that Sp1 transcription factor mediates constitutive and TGF-beta-inducible expression of the huPAR gene in U937 cells through binding to the sequence located at -70. PMID- 10684976 TI - Cloning and characterization of additional members of the G protein-coupled receptor family. AB - A search of the expressed sequence tag (EST) database retrieved a human cDNA sequence which partially encoded a novel G protein-coupled receptor (GPCR) GPR26. A human genomic DNA fragment encoding a partial open reading frame (ORF) and a rat cDNA encoding the full length ORF of GPR26 were obtained by library screening. The rat GPR26 cDNA encoded a protein of 317 amino acids, most similar (albeit distantly related) to the serotonin 5-HT(5A) and gastrin releasing hormone BB2 receptors. GPR26 mRNA expression analysis revealed signals in the striatum, pons, cerebellum and cortex. HEK293 and Rh7777 cells transfected with GPR26 cDNA displayed high basal cAMP levels, slow growth rate of clonal populations and derangements of normal cell shape. We also used a sequence reported only in the patent literature encoding GPR57 (a.k.a. HNHCI32) to PCR amplify a DNA fragment which was used to screen a human genomic library. This resulted in the cloning of a genomic fragment containing a pseudogene, psiGPR57, with a 99.6% nucleotide identity to GPR57. Based on shared sequence identities, the receptor encoded by GPR57 was predicted to belong to a novel subfamily of GPCRs together with GPR58 (a.k.a. phBL5, reported only in the patent literature), putative neurotransmitter receptor (PNR) and a 5-HT(4) pseudogene. Analysis of this subfamily revealed greatest identities (approximately 56%) between the receptors encoded by GPR57 and GPR58, each with shared identities of approximately 40% with PNR. Furthermore, psiGPR57, GPR58, PNR and the 5-HT(4) pseudogene were mapped in a cluster localized to chromosome 6q22-24. PNR and GPR58 were expressed in COS cells, however no specific binding was observed for various serotonin receptor-specific ligands. PMID- 10684977 TI - Transcription factor GATA-6 activates expression of gastroprotective trefoil genes TFF1 and TFF2. AB - One of the early events in inflammation and epithelial restitution of the gastrointestinal tract is the up-regulation of secretory peptides belonging to the trefoil factor family (TFF) that promote cell migration, protect and heal the mucosa. Their major expression site is stomach (TFF1, TFF2) and intestine (TFF3). Located in the 5'-flanking region of the genes are several consensus sites for members of the GATA transcription factors known to control gut-specific gene expression. By reverse transcription-PCR (RT-PCR), GATA-6 was shown to be expressed in a variety of tumor cell lines of gastric, intestinal and pancreatic origin. In MKN45, KATOIII and LS174T, cotransfection with TFF reporter genes and GATA-6 expression vectors revealed that GATA-6 activates TFF1 and TFF2 4-6-fold, without an effect on TFF3. The functional contribution of GATA binding sequences in the reverse orientation was further characterized by reporter gene assays using TFF2 deletion constructs and by gel shift experiments. PMID- 10684978 TI - Complete sequence and characterization of chick ventricular myosin heavy chain in the developing atria. AB - We isolated five complementary DNA (cDNA) clones, encoding the chick ventricular myosin heavy chain (MyHC) by reverse transcription polymerase chain reaction (RT PCR). The entire cDNA consists of 5995 nucleotides with the 52 bp 5'-untranslated region and the 129 bp 3'-untranslated region. The complete cDNA encodes 1937 amino acids. Expression of the chick ventricular MyHC gene was also studied by Northern blot analysis. This gene continued to be strongly expressed in the ventricle during cardiac development. On the other hand, its expression was moderate in the early embryonic atria, and was down-regulated during development. In the adult atria, this gene was expressed at very low levels. To determine the localization of the ventricular MyHC protein, an immunohistochemical study was performed. The ventricular MyHC was present in early embryonic atrial myocytes. During development, the expression of this protein in the atrial myocytes was down-regulated, but continued to be present in the atrial conduction system. Our results indicate that the ventricular MyHC appears in the primary atrial myocardium and is then localized in the conduction cells of the atria. PMID- 10684979 TI - Expression of CD80 promoter in transgenic mice. AB - CD80 is a very potent co-stimulatory factor which is required for complete T-cell activation. Here, we use transgenic mice as a tool to map the promoter of the CD80 gene. We engineered three different CD80 promoter driven luciferase transgenes: -3084, -1073 and -215. With these transgenes, we have generated three groups of transgenic mice. Our results showed that the -3084 CD80 promoter/luciferase transgene was sufficient to confer tissue-specific expression of the CD80 gene. When the promoter sequence was deleted to -1073, the normal tissue-specific expression was lost. A brain-specific element was mapped between 1073 nt and -215 nt. This element caused up to ninefold higher expression of the CD80 promoter/luciferase in brain tissue of -1073 CD80 promoter/luciferase transgenic animals as compared to -3084 CD80 promoter/luciferase transgenic animals. In contrast to results with a cell culture system, little luciferase activity was detected in -215 CD80 promoter/luciferase transgenic animals. PMID- 10684980 TI - Transcriptional regulation involving the intronic heat shock element of the rat hsp27 gene. AB - While sequencing the first intron of the rat heat shock protein 27 gene (hsp27), we identified a consensus heat shock regulatory element (HSE). This intronic HSE (i-HSE) is conserved among mammalian hsp27 genes. The aim of this study was to investigate possible effects of this intronic HSE (i-HSE) on transcription from the rat hsp27 promoter. Gel mobility shift assays indicated that the i-HSE bound heat shock transcription factor 1 (HSF1) in a manner equivalent to that of HSE present in hsp27 promoter (p-HSE). The effect of i-HSE on transcription from the hsp27 promoter was evaluated using reporter constructs transiently transfected in the osteosarcoma cell line ROS17/2.8. When inserted 5' to a 145 bp fragment of the hsp27 promoter not containing p-HSE, a 215 bp fragment of hsp27 intron 1 containing i-HSE enhanced CAT activity and conferred heat shock-inducible activity to the construct. This intronic fragment containing i-HSE also enhanced CAT activity in either normal or heat-shocked culture conditions when inserted 3' to the CAT open reading frame. However, in chimeric reporter constructs with a 273 bp hsp27 promoter containing p-HSE directly 5' to CAT reporter, and with a 215 bp fragment containing i-HSE inserted 3' to the CAT open reading frame, transcription from hsp27 promoter was reduced under normal and heat-stressed culture conditions. Mutation of the i-HSE reversed this effect. Further study is required to define the mechanism by which the i-HSE-containing region of the hsp27 promoter may mediate negative regulation of hsp27 transcription. PMID- 10684981 TI - A gene cluster encoding plantaricin 1.25beta and other bacteriocin-like peptides in Lactobacillus plantarum TMW1.25. AB - Plantaricin 1.25beta is a thermostable class two bacteriocin produced by Lactobacillus plantarum TMW1.25 isolated from sausage fermentation. It is co produced with several other bacteriocin-like peptides. Using oligonucleotides derived from previously determined peptide sequences, a 3.8 kb DNA fragment could be amplified. A neighboring 1.8 kb fragment was amplified using ligation-anchored single-specific-primer PCR. Sequencing of the complete 5.6 kb stretch revealed that the structural gene for plantaricin 1.25beta, plnB, was located downstream of another bacteriocin gene, plnC. Seven other open reading frames were detected, including plnK encoding a bacteriocin-like peptide, but not including any putative immunity genes. Interestingly, the gene cluster contained an IS30-like insertion sequence, designated IS125, as well as an ISS1 homolog. PMID- 10684982 TI - Structural conservation and variation among U5 small nuclear RNAs from trypanosomatid protozoa. AB - U5 snRNAs in trypanosomatid protozoa do not contain the trimethylguanosine cap structures that are often targeted in snRNA isolation procedures. As a result, the trypanosomatids are not well represented in the database of available U5 snRNA sequences. We have isolated and determined the sequence of the U5 snRNA from Crithidia fasciculata. Comparison with previously published trypanosomatid U5 snRNA sequences allows us to deduce the pattern of structural conservation and variation among these very divergent snRNA molecules. PMID- 10684983 TI - Corrigendum to 'Molecular cloning and expression of human neurochondrin-1 and 2'(1). PMID- 10684984 TI - Androstenediol stimulates myelopoiesis and enhances resistance to infection in gamma-irradiated mice. AB - The ionizing radiation-induced hemopoietic syndrome is characterized by defects in immune function and increased mortality due to infections and hemorrhage. Since the steroid 5-androstene-3beta, 17beta-diol (5-androstenediol, AED) modulates cytokine expression and increases resistance to bacterial and viral infections in rodents, we tested its ability to promote survival after whole-body ionizing radiation in mice. In unirradiated female B6D2F1 mice, sc AED elevated numbers of circulating neutrophils and platelets and induced proliferation of neutrophil progenitors in bone marrow. In mice exposed to whole-body (60)Co gamma radiation (3 Gy), AED injected 1 h later ameliorated radiation-induced decreases in circulating neutrophils and platelets and marrow granulocyte-macrophage colony forming cells, but had no effect on total numbers of circulating lymphocytes or erythrocytes. In mice irradiated (0, 1 or 3 Gy) and inoculated four days later with Klebsiella pneumoniae, AED injected 2 h after irradiation enhanced 30-d survival. Injecting AED 24 h before irradiation or 2 h after irradiation increased survival to approximately the same extent. In K. pneumoniae-inoculated mice (irradiated at 3-7 Gy) and uninoculated mice (irradiated at 8-12 Gy), AED (160 mg/kg) injected 24 h before irradiation significantly promoted survival with dose reduction factors (DRFs) of 1.18 and 1.26, respectively. 5-Androstene-3beta ol-17-one (dehydroepiandrosterone, DHEA) was markedly less efficacious than AED in augmenting survival, indicating specificity. These results demonstrate for the first time that a DHEA-related steroid stimulates myelopoiesis, and ameliorates neutropenia and thrombocytopenia and enhances resistance to infection after exposure of animals to ionizing radiation. PMID- 10684985 TI - Preliminary studies on the immunomodulatory effect of the C3 binding glycoprotein isolated from Cuscuta europea. AB - This study investigates the immunomodulatory effect of a C3 binding glycoprotein (C3bgp), isolated from the parasitic plant Cuscuta europea. When BALB/c mice, immunized with sheep red blood cells (SRBC), were given a single intraperitoneal injection of C3bgp a dose-dependent immunostimulation was observed. The stimulation was assessed by an increase in the number of haemolytic plaque forming cells (PFC) and haemaglutination titres. The induction was time dependent in respect to the administration of both the C3bgp and SRBC. When C3bgp was applied 24 h before SRBC at a dose of 30 microg per mouse (1.2 mg/kg), a well expressed immunostimulation was found. It was also found that giving C3bgp to mice, which had previously been treated with the immunosuppressive drug cyclophosphamide (CY), produced an increase in PFC. The immune response was also restored in vitro experiments were performed using human whole blood cultures stimulated with 30 microg/ml C3bgp in the presence or absence of egg albumin (OVA) as antigen for 72 to 168 h. In C3bgp stimulated cultures it was found that after 120 h there was a high expression of the CD 19+ subset of the activation antigen CD25 (IL-2R) as assessed by flow cytometric phenotype analysis. Supernatants from cultures with different stimuli were assayed by a solid phase ELISA for the determination of OVA-specific IgM at 120, 144 and 168 h. It was found that C3bgp application alone, failed to enhance OVA specific IgM, but significantly high levels of IgM in cultures containing C3bgp and OVA, were detected. Overall it has been shown that the C3 binding glycoprotein, as obtained from the parasitic plant Cuscuta europea, has strong immunostimulatory properties both in vivo and in vitro. PMID- 10684986 TI - The effect of the antithyroid drug propylthiouracil on the alternative pathway of complement in rats. AB - The effect of propylthiouracil (PTU) on the lytic activity of complement in rat serum was investigated in vivo. Rats (180+/-10 g) were treated daily by gavage with PTU doses of 1-50 mg/200 g body weight for time intervals ranging from 1 to 30 days. Serum classical pathway (CP) and alternative pathway (AP) activities were determined 24 h after the last dose. A single dose of 50 mg/200 g body weight was administered to additional groups and the animals were sacrificed after periods of 1-48 h. The results showed a relatively small reduction ( approximately 30%) in CP activity, evident only in animals treated with 50 mg of PTU for three weeks. However, a clear and opposite effect of PTU, an increase in lytic activity reaching values up to 180% of controls, was observed on AP activity. This effect was seen at all PTU doses used, and occurred within 4 days of treatment with the highest dose. Maximum activity was observed at intermediate intervals, depending on the PTU dose, with a return to control levels occurring after the longer periods of treatment. The lytic activity of serum from animals treated with a single PTU dose of 50 mg/200 g body weight and sacrificed 1-48 h after dosing did not differ from controls. Serum levels of thyroid hormone (triiodo L-thyronine, T3, and thyroxine, T4) were determined in representative groups of treated animals (injected with 5 mg of PTU/200 g body weight/day). These were either undetectable or considerably lower than those of controls. The serum PTU levels of these rats increased for up to 22 days, reaching values of 2 4 microg/ml.PTU is described in the literature as a modulator of both cellular immune responses and antibody production. Upon complement activation fragments of complement components bind to immune complexes and to specific receptors on cells of the immune system. Thus, alteration in AP activity caused by PTU treatment suggests a possible mechanism by which the drug exerts its modulatory effect. Increased complement AP activity might affect events as antigen presentation and hence the onset and course of the immune response. PMID- 10684987 TI - Augmentation by interleukin-18 of MHC-nonrestricted killer activity of human peripheral blood mononuclear cells in response to interleukin-12. AB - Interleukin (IL)-18 is a novel cytokine with pleiotropic functions. In the present study, we examined the induction of the killer activity of peripheral blood mononuclear cells (MNC) against lung cancer cell lines upon treatment with IL-18 in combination with IL-12. Cytotoxic activity was measured by standard (51)Cr release assay. IL-18 (100 ng/ml) was found to significantly augment IL-12 induced killer activity in a MHC-nonrestricted manner against allogeneic NK resistant Daudi cells and lung cancer cell lines: SBC-3, RERF-LC-AI and A549. IL 18 could augment IL-12-induced killer activity both at the optimal as well as suboptimal doses of the latter. However, IL-18 was found to have little effect on the killer activity of MNC induced by optimal or suboptimal dose of IL-2 or IL 15. Treatment of MNC with IL-18 in combination with IL-12 for a period of more than 4 days was observed to optimally induce the killer activity. As for induction of IFN-gamma production by MNC, IL-18 augmented that induced by IL-2 and IL-15, as well as that induced by IL-12. These results show the potential of IL-18 in combination with IL-12 for clinical application in treatment of cancer. PMID- 10684988 TI - Protected Trypanosoma cruzi infection in rats born to mothers receiving interferon-gamma during gestation is associated with a decreased intramacrophage parasite growth and preferential synthesis of specific IgG2b antibodies. AB - We demonstrated that administration of interferon gamma (IFN-gamma) to pregnant rats conferred partial resistance in their offspring to further challenge with Trypanosoma cruzi. Because of the effects of IFN-gamma on macrophage activation and immunoglobulin isotype selection, offspring were now studied to ascertain whether this intervention modifies the in vitro replication of T. cruzi and nitric oxide (NO) production by peritoneal macrophages (PE), together with the anti-T. cruzi IgG isotypes. To evaluate the possibility of a detrimental effect of IFN-gamma, serum levels of anti-sulphatide autoantibodies were also investigated. Offspring were born to mothers undergoing one of the following procedures during gestation: treatment with recombinant rat IFN-gamma, 50,000 IU/rat, five times/week for 3 weeks, which was started on the day of mating; infection with 10(6) trypomastigotes of T. cruzi at 7, 14, and 21 days after mating plus IFN-gamma treatment as given to the former group; the same protocol except that physiological saline was injected instead of IFN-gamma; injection of physiological saline only. Offspring were challenged at weaning with a similar dose of T. cruzi, to constitute four groups of infected young, plus an additional group of age-matched uninfected rats born to control mothers. PE were harvested at day 7 postinfection (pi), exposed to parasites and further investigated for the replication of T. cruzi and NO production, whereas ELISA studies for measuring serum anti-T. cruzi IgG subclasses and anti-sulphatide autoantibodies were performed at day 30 pi. The number of intracellular parasites in PE was markedly decreased in young born to IFN-gamma-treated mothers, this not being accompanied by higher nitrite levels in culture supernatants. Offspring delivered by IFN-gamma-treated mothers showed no higher serum concentrations of anti sulphatide autoantibodies, but exhibited a preferential synthesis of anti-T. cruzi IgG2b antibodies. This rat isotype is known to fix complement and constitutes the rat counterpart of IgG2a mouse immunoglobulins whose synthesis is favoured by IFN-gamma. PMID- 10684989 TI - Immunogenicity of an E. coli extract after oral or intraperitoneal administration: induction of antibodies against pathogenic bacterial strains. AB - For the treatment of recurrent infections of the urinary tract, a bacterial extract (OM-89) consisting of immunostimulating components derived from 18 Escherichia coli strains is orally applied to patients. We investigated in a mouse model the immunogenicity of the bacterial extract after intraperitoneal or oral administration. After repeated administration of the extract, serum IgG and IgA responses against the E. coli strains used for the preparation of OM-89 were obtained. This antisera also recognized a number of bacterial strains isolated from patients with urinary tract and enterohemorrhagic E. coli infections, and bound to a variety of other pathogenic strains. Moreover, the supernatants of cell cultures prepared from the urogenital tract of mice immunized with OM-89 contained increased levels of strain specific and of total IgG and IgA. Our findings may contribute to explain the therapeutic effect of OM-89 demonstrated in clinical studies. PMID- 10684990 TI - Effects of imipenem and cilastatin on human T-lymphocytes derived from acute leukemia patients with chemotherapy-induced leucopenia: studies of T-lymphocyte responses in the presence of acute myelogenous leukemia (AML) blast accessory cells. AB - The effects of imipenem and cilastatin on human T-lymphocytes were studied in vitro. As responder T-cells were used T-lymphocyte clones derived from acute leukemia patients with chemotherapy-induced cytopenia, and the accessory cells were highly enriched acute myelogenous leukemia (AML) blasts. The effects of imipenem and cilastatin on phytohemagglutinin (PHA), anti-CD3 and anti-CD3+anti CD28 stimulated activation were assayed, and in addition drug effects on cytokine dependent proliferation of activated T-lymphocytes were investigated. Imipenem inhibited IL2-dependent proliferation of activated CD4(+) and CD8(+) T-cell clones, and an inhibition was also detected for IL7-, IL12-, IL15-, IL16- and IL17-dependent clonal proliferation. Imipenem caused a weak inhibition of anti CD3- and PHA-stimulated T-cell proliferation when using 50 Gy irradiated AML blast accessory cells derived from various patients, whereas no effect was observed for anti-CD3+anti-CD28 stimulated and allostimulated activation. Imipenem decreased the release of IL4 and Interferon-gamma by T-cell clones stimulated with anti-CD3 and PHA in the presence of native (nonirradiated) AML blasts. The imipenem effects were observed at concentrations corresponding to levels reached in vivo, whereas even high concentrations of cilastatin did not alter T-cell responses. The T-lymphocyte inhibition is probably caused by a direct effect of imipenem on the T-cells. PMID- 10684991 TI - Influence of pyrrolidine dithiocarbamate on the inflammatory response in macrophages and mouse endotoxin shock. AB - To analyze the immunomodulatory effect of pyrrolidine dithiocarbamate (PDTC) on the endotoxin (LPS) stimulated inflammatory response, we measured the LPS stimulated cytokine and NO production in murine peritoneal macrophages, J774A.1 cells and human whole blood in the presence of PDTC (60 microM). PDTC significantly inhibited the production of nitrite, IL-1beta and IL-6 in these cells. TNFalpha release was stimulated in murine cells, but suppressed in human whole blood. We further investigated the influence of PDTC on mortality and cytokine release in mouse endotoxin shock. PDTC was i.p. injected 30 min prior to the induction of endotoxin shock in female NMRI-mice and survival was significantly improved as compared to controls (48% vs 20%, n=25 per group). Plasma concentrations of TNFalpha were slightly augmented while IL-6 levels were decreased in PDTC-treated animals as compared to controls, however, without reaching significance. We conclude that PDTC is a potent immunomodulatory substance that modulates the inflammatory response in vitro and reduces mortality in mouse endotoxin shock. The pathophysiological mechanisms of the protective effect of PDTC in vivo, however, appears to be pluripotent, comprising both antioxidative properties and the inhibition of NF-kB. PMID- 10684992 TI - Adamantylamide dipeptide stimulates hematopoiesis and increases survival in irradiated mice. AB - It has been demonstrated that the synthetic immunostimulatory compound, adamantylamide dipeptide (AdDP) produces hematopoiesis-stimulating effects in mice exposed to sublethal doses of ionizing radiation and increases survival in experimental animals irradiated with a lethal dose. These findings might suggest contingent extension of clinical indications for the administration of AdDP for the conditions of hematopoietic suppression, especially in oncology. PMID- 10684993 TI - Editorial PMID- 10684994 TI - Inhibition of influenza virus replication by cocaine. AB - Cocaine has been shown to have a number of diverse effects on the immune system. The current investigators have previously demonstrated an inhibitory effect of cocaine on murine hepatitis virus replication in peritoneal macrophages in vitro. The present study was undertaken to examine the effects of cocaine on influenza virus replication and to further characterize that effect in an animal model. Cocaine was capable of inducing a dose-dependent reduction in influenza PR-8 replication using MDCK cells in vitro. Concentrations of 100 microg/ml caused a 50% reduction of virus. To further characterize the effect in vivo, C57Bl/6 mice infected with influenza PR-8 by intranasal instillation were given daily ip injections of 10 mg/kg cocaine just prior to and for 4 days after exposure to influenza. Lungs from mice exposed to cocaine had viral titers that were reduced approximately 50% compared to controls as demonstrated by hemagglutination titers. Additional studies suggest that this reduction appears to be caused by an increase of cocaine-induced interferon. PMID- 10684995 TI - B cell stimulating activity of metallothionein in vitro. AB - The effect of metallothionein (MT) on lymphocytes was studied in vitro. Rabbit MT induced the proliferative responses of mouse splenocytes at concentrations of 1 25 microg/ml. MT synergistically enhanced Con A- or LPS-induced proliferative response of splenocytes. A similar effect was also observed for rabbit splenocytes at a similar concentration. Free heavy metals such as Cd and Zn only weakly stimulated splenocytes. 2-mercaptoethanol (2-Me) abrogated the effect of MT, suggesting that thiols in MT play an important role for splenocyte response. MT stimulated splenocytes from MT-knockout mice as well as those from normal control mouse. The responder cells for MT stimulation were B cells and MT also induced B cell differentiation to produce Ig. MT induced the calcium influx of B cells, but not T cells. These results indicate that MT has a potent immunomodulating activity, especially on B cells. PMID- 10684996 TI - Inhibitory effect of TMK-688 on late asthmatic responses as well as T-cell and eosinophilic infiltration in guinea pigs with asthmatic reactions. AB - The effects of an oral anti-allergic agent, TMK-688, which inhibits 5 lipoxygenase, at doses of 3.2 and 10 mg/kg were studied in guinea pigs with dual phase asthmatic response. We previously observed that pretreatment with TMK-688 inhibited the late asthmatic response (LAR) induced by ovalbumin inhalation exposure. The present study focused on the effect of TMK-688 on infiltration by T cells and eosinophils. TMK-688 inhibited both T-cell and eosinophilic infiltration. These findings suggest that TMK-688 is effective in inhibiting infiltration of T-cells and eosinophilic chemotaxis, and thereby suppresses LAR. PMID- 10684997 TI - In vitro IgE inhibition in B cells by anti-CD23 monoclonal antibodies is functionally dependent on the immunoglobulin Fc domain. AB - CD23, the low affinity receptor for IgE (FcvarepsilonRII), is involved in regulation of IgE synthesis by B-lymphocytes. Five monoclonal antibodies to human CD23 were generated from cynomolgus macaques immunized with purified soluble CD23 (sCD23). Four of the five primate antibodies blocked the binding of IgE complexes to CD23 positive cells and also inhibited the production of IgE in vitro by IL-4 induced human peripheral blood mononuclear cells (PBMC). The variable domains of several primate antibodies were utilized to construct chimeric macaque/human (PRIMATIZED((R))) monoclonal antibodies. PRIMATIZED((R)) p5E8G1, containing human gamma 1 constant region, inhibited IgE production in vitro as efficiently as the parent primate antibody, but the human gamma 4 constant version, PRIMATIZED((R)) p5E8G4, was not as effective in IgE inhibition. An F(ab')(2) of p5E8G1 did not inhibit IgE production but did interfere with IgE inhibition by the intact anti CD23 antibody in a dose dependent fashion. The murine monoclonal antibody MHM6 recognizes human CD23 at a different epitope than primate antibody 5E8, and inhibits IgE production by IL-4 induced PBMC. As with the F(ab')(2) of p5E8G1, the F(ab')(2) of MHM6 also failed to inhibit IgE production. These data imply that the mechanism by which anti-CD23 antibodies inhibit IgE production requires cross-linking of CD23 to an IgG receptor. These data also imply that neither bivalent cross-linking of CD23 alone or inhibition of CD23 binding to its natural ligands is sufficient to inhibit IgE production. PMID- 10684998 TI - Phenytoin and electric shock-induced apoptosis in rat peripheral blood lymphocytes. AB - The apoptotic index (AI) of peripheral blood lymphocytes (PBL) and plasma corticosterone (CS) levels were determined in Wistar rats treated with phenytoin (PHT) at therapeutic and toxic doses (100 or 200 mg/kg/day, respectively, over a period of 7 days) and stressed by bifrontal electric shock (60 Hz/40 mA/0.2 seg). The values of CS and AI were found to be significantly higher in rats submitted to electric shock (ES) and in rats treated with therapeutic and toxic doses of PHT plus ES, than in rats treated only with PHT (P<0.001). The plasma concentrations of PHT were found to be significantly higher in rats treated with toxic doses than in those treated with therapeutic doses (P<0.001), while the control group (without treatment) and vehicle group (propilenglycol-ethanol water, 40:10:50), showed low levels of CS, and less than 1% of AI. The DNA analysis by electrophoresis in agarose in all the groups was positive, displaying the ladder pattern characteristic of apoptotic process (200 bp), except in the control groups (no treatment and vehicle treated). Our results demonstrate that chronic stress, caused by ES, produces an elevation of CS. The values of apoptosis were correlated with the CS levels, suggesting that the apoptotic inductor process is a consequence of an increase in the concentration of corticosterone in plasma, in response to the hypothalamic-pituitary-adrenals (HPA) axis activation, while phenytoin at therapeutic doses is only a moderate apoptosis inductor. PMID- 10684999 TI - Nitrite inhalants spontaneously liberate nitric oxide, which is not responsible for the immunotoxicity in C57BL/6 mice. AB - Nitrite inhalant abuse has been correlated epidemiologically with HIV seropositivity and with Kaposi's sarcoma. Using a mouse model, we have shown that inhaled isobutyl nitrite caused anemia and severely depressed immunity. In the present study, we showed that both isobutyl and cyclohexyl nitrites in air liberated nitric oxide (NO). An immunotoxic dose of 900 ppm isobutyl nitrite liberated 115 ppm NO. Mice were exposed in an inhalation chamber to 115 ppm NO, 900 ppm isobutyl nitrite, or 900 ppm cyclohexyl nitrite for 45 min/day. Following a single exposure, NO did not affect peripheral blood cell counts, while isobutyl and cyclohexyl nitrites reduced cell numbers. After 14 daily exposures, isobutyl nitrite, but not cyclohexyl nitrite or NO, reduced peritoneal macrophage tumoricidal activity. The nitrite esters likely caused immunotoxicity by mechanisms other than NO release. PMID- 10685000 TI - Gestational nicotine exposure alone or in combination with ethanol down-modulates offspring immune function. AB - Prenatal nicotine exposure has been shown to disrupt the development of a number of peripheral organs. In the current study, we examined the effects of gestational nicotine exposure, alone or in combination with ethanol exposure, on offspring immune function. Timed pregnant rats were treated with either nicotine (6 mg/kg/day) from gestation day 4-20 using subcutaneously implanted osmotic mini pumps or ethanol administered in the drinking water (15% w/v) from gestation day 10-20. The combined exposure group received both treatments. The ability of offspring T and B cells to proliferate in response to nonspecific stimulation by Concanavalin A or lipopolysaccharide, respectively, was determined on postnatal days 9, 15, 22, 29, 64, and 86. Offspring splenocyte beta(2)-adrenoceptor binding was also measured. Nicotine or nicotine+ethanol suppressed splenocyte responsiveness to Concanavalin A or lipopolysaccharide which was similar in timing and magnitude to that seen with ethanol alone. Splenocytes from these groups remained subresponsive to stimulation well into adulthood. The combined drug treatment caused an overall reduction in spleen beta-adrenergic receptor binding whereas the individual drug treatments did not alter the development of spleen beta-adrenergic receptors.Our results indicate that prenatal nicotine exposure can cause long-term suppression of the proliferative response of offspring immune cells. Moreover, the effects of nicotine+ethanol may cause more severe deficits in adulthood. PMID- 10685002 TI - FK506 inhibition of histamine release and cytokine production by mast cells and basophils. AB - Histamine release and cytokine production by mast cells and basophils are thought to be closely involved in the pathogenesis of allergic diseases. Some reports show that FK506 (tacrolimus hydrate) inhibited histamine release and cytokine production by mast cells and basophils. However, as the effects of FK506 has not been compared with those of clinically used drugs in those reports, the clinical relevancy of FK506 inhibition remained unclear. In this paper, we compared the actions of FK506 with those of steroids or disodium cromoglycate (DSCG) which has been clinically used. FK506 inhibited histamine release by Brown-Norway rat peritoneal mast cells more potently than steroids and especially DSCG. FK506 also inhibited histamine release by a mast rat basophilic leukemia (RBL)-1 cell line and human peripheral blood basophils, whereas steroids failed to inhibit histamine release by human basophils. FK506 as well as steroids inhibited TNF alpha and IL-4 production by RBL-1 cells. FK506 was therefore more effective than steroids and DSCG in inhibiting histamine release, and it also had the ability of inhibiting cytokine production by mast cells as steroids do. We concluded that FK506 might regulate allergic diseases via these actions, judging from the viewpoint of clinical relevancy. PMID- 10685001 TI - Cytokines potentiate human eosinophil superoxide generation in the presence of N(omega)-nitro-L-arginine methyl ester. AB - The eosinophilic (EOS) leukocyte has been implicated as a primary effector cell in inflammatory and allergic diseases. Cytokines are among the mediators of inflammatory and allergic diseases which modulate the effector functions of EOS. Certain cytokines, elevated in patients with various allergies, are thought to modulate EOS reactive oxygen species superoxide anion and nitric oxide (NO) responses. Though EOS transcribe and translate mRNA for inducible NO synthase, the effects of cytokines on NO generation remain largely unknown. Thus, we have investigated effects of IL-3, IL-5, GM-CSF, IL-8, RANTES and the proinflammatory cytokines TNF-alpha and IFN-gamma, on superoxide anion and NO generation by clone 15 HL-60 human eosinophilic cells. Cytokine treatments (3 and 18 h) resulted in production of small amounts of superoxide anion which were enhanced by the NO inhibitor L-NAME. In the presence of L-NAME, PMA (1 nM) stimulation significantly increased superoxide anion generation following 3 h treatments with IL-3, TNF alpha or IFN-gamma. Eighteen hour cytokine treatments with GM-CSF, IL-8, RANTES, IFN-gamma or TNF-alpha primed the cells for enhanced reactive oxygen species following exposure to an EOS stimulant. Inhibition of NO synthesis resulted in increased levels of superoxide anion. Collectively, these results suggest that an environment of proinflammatory cytokines may potentiate the generation of reactive oxygen species by EOS. These results further suggest that at an inflammatory site or during an allergic response, EOS may concomitantly synthesize NO and generate superoxide anion, fractions of which may rapidly react to form the potent oxidant peroxynitrite. PMID- 10685003 TI - Modulatory effect of 7-thia-8-oxoguanosine on proliferation of rat thymocytes in vitro stimulated with concanavalin A. AB - 7-thia-8-oxoguanosine (immunosine) is a guanosine analogue showing immunostimulatory activity on different components of the immune system, including B lymphocytes, natural killer cells and macrophages. However, little is known about its effect on T-cell functions. In this work it was demonstrated that immunosine at concentrations between 10 microM and 1 mM stimulated proliferation of rat thymocytes in vitro triggered by suboptimal concentrations of concanavalin A (Con A). The effect correlated with increased interleukin 2 (IL-2) production, upregulation of the IL-2 receptor alpha (IL-2Ralpha) expression and decreased apoptosis of thymocytes in comparison to the effect of Con A alone. PMID- 10685004 TI - Increased T cell cytotoxicity by Betathine-induced upregulation of TNFalpha. AB - Betathine (BT) is a low molecular weight disulfide that has previously been shown to exhibit in vivo antitumor activity in murine myeloma and melanoma models. We have shown that BT treatment of both human T cells and monocytes is associated with an increase in surface tumor necrosis alpha (TNFalpha) expression. Further, in T cells and monocytes that have been stimulated with PMA and ionomycin, the addition of BT results in a dose and time dependent increase in the percentage of high TNFalpha-expressing cells. Unlike TNFalpha upregulation produced by the commonly used thiol antioxidant N-acetyl-L-cysteine (NAC), the BT-induced increase in TNFalpha is observed consistently in different donors. This increase in surface TNFalpha is associated with elevated levels of TNFalpha mRNA. In addition, expression of TNFalpha receptor I is also significantly enhanced by BT treatment. The upregulation of surface TNFalpha by BT has functional consequences, in that, BT-treated T cells exhibit enhanced cytotoxic activity. Thus, increased TNFalpha expression may be one mechanism responsible for the antineoplastic activity of BT. PMID- 10685005 TI - Evidence for immunotoxic effects of crude Ginkgo biloba L. leaf extracts using the popliteal lymph node assay in the mouse. AB - Allergic reactions due to contact with different parts of the ancient tree Ginkgo biloba L. have repeatedly been reported. Provocation tests in patients and animal experiments have identified alkylphenols such as ginkgolic acids as causative constituents. Leaf extracts from Ginkgo are widely used to treat peripheral or cerebral circulatory disorders and Alzheimer's disease. Since alkylphenols are also present in leaves, potential allergic and other immunological hazards of such preparations have to be carefully controlled. Thus, we have evaluated if the popliteal lymph node assay (PLNA) in the mouse may represent a suitable model for the detection of constituents with immunotoxic properties in a complex mixture of biologically active agents such as plant extracts. Subplantar injection (2 mg) of a crude aqueous-ethanolic extract from Ginkgo leaves caused a significant lymphoproliferative reaction (LPR) in the ipsilateral popliteal lymph node. PLNA active compounds in this extract could be enriched in the lipophilic phase by liquid-liquid partition between heptane and water. Chemical analysis of the heptane extract revealed the presence of a high concentration of alkylphenols (approx. 30%) and further subfractionation indicated that the enlargement of the popliteal lymph node was mainly due to the content of ginkgolic acids. This presumption was corroborated by observing a similar LPR following injection of a purified mixture of ginkgolic or hydroginkgolic acids. Thus, our experiments confirm that Ginkgo leaf extracts may contain constituents with immunotoxic properties, underlining the need to apply adequate production procedures to guarantee the completest possible removal of these compounds. The PLNA appears to represent a simple test model for the detection, characterisation and control of ingredients with potential immunotoxic side effects in complex herbal drugs. PMID- 10685006 TI - Aflatoxin B(1) inhibits CD14-mediated nitric oxide production in murine peritoneal macrophages. AB - Aflatoxin B(1) (AFB(1)), a potent hepatocarcinogen, has been known to impair non specific and specific immunity. Macrophages play an important role in host defense against tumors and microorganisms and a number of compounds are implicated in macrophage cytotoxicity. Since activated by the reaction of LPS with CD14, macrophages produce nitric oxide (NO) that is a cytotoxic effector molecule in cell killing. In the present study, we investigated whether the alteration of CD14 level on macrophages by AFB(1) affects NO production in murine peritoneal macrophages. When macrophages were stimulated with LPS after AFB(1) pretreatment, or they were co-treated with LPS and AFB(1), the NO production decreased in a dose-dependent manner. In contrast, when macrophages were post treated with AFB(1) after LPS-stimulation, NO production was unchanged. DNA, RNA, and protein synthesis were reduced by AFB(1)-pretreatment of macrophages. The addition of anti-CD14 antibodies to the cultures decreased NO production further. FACS analysis showed that the binding of anti-CD14 antibodies to the macrophages was suppressed by AFB(1)-pretreatment followed by LPS-stimulation. However, AFB(1) does not alter the binding anti-CD14 antibodies to the macrophages without LPS-stimulation. In contrast, AFB(1) pretreatment increased an amount of CD14 released in culture medium. Taken together, these data indicate that the reduced NO production in murine peritoneal macrophages by AFB(1)-pretreatment is related to the suppressed expression of CD14 on macrophage membrane and to the increased secretion of it to culture medium after LPS-stimulation. PMID- 10685007 TI - Strain differences in binding properties of estrogen receptors in immature and adult BALB/c and MRL/MP-lpr/lpr mice, a model of systemic lupus erythematosus. AB - The aim was to compare binding properties of estrogen receptors in brain, reproductive and immune tissues of immature and adult female BALB/c mice, and in the same tissues of MRL/MP-lpr/lpr mice. The latter strain spontaneously develops an autoimmune disease resembling human systemic lupus erythematosus (lupus; SLE). It is hypothesized that estradiol, through its receptors, mediates the progression of murine SLE. High-speed cytosols were prepared from hypothalamus, spleen, thymus and uterus of both strains, and incubated with the synthetic estrogen (3)H-moxestrol (NEN). Scatchard plots were derived from binding isotherms obtained after in vitro incubation. In addition, cervical lymph nodes from MRL mice could be used, but were too small in BALB/c mice. There was a significant increase in the affinity of the binding reaction i.e. a decrease in the apparent molar dissociation constant (Kd), in immune tissues and uterus with maturation in MRL but not BALB/c mice, whose tissues had, overall, a lower affinity for (3)H-moxestrol. Receptor concentrations were significantly higher in spleen and cervical lymph nodes of adult compared with immature MRL mice, but the opposite pattern was observed in BALB/c mouse spleen on maturation. These properties of estrogen receptors in MRL mice may underlie estrogen-mediated exacerbation of murine SLE. PMID- 10685008 TI - Mutagenicity and antimutagenicity studies of tannic acid and its related compounds. AB - Tannic acid and its hydrolysed products such as ellagic acid, gallic acid and propyl gallate were tested for mutagenicities using Ames Salmonella tester strains TA98 and TA100. Also, the antimutagenic activities of these compounds against a number of direct mutagens including 2-nitrofluorene (2-NF), 4,4' dinitro-2-biphenylamine, 1-nitropyrene, 1,3-dinitropyrene, 2-nitro-p phenylenediamine, 3-nitro-o-phenylenediamine, 4-nitro-o-phenylenediamine were tested. None of these tannic acid compounds was mutagenic. They also failed to show antimutagenic activity towards the tested direct mutagens. However, tannic acid at non-growth inhibitory concentrations reduced the revertant numbers of TA98 in the presence of S9 mix when benzidine, 3,3'-4,4'-tetraminobiphenyl, 4 aminobiphenyl, and N,N-N', N'-tetramethylbenzidine were used as the mutagens. These results suggest that tannic acid, but not its hydrolytic products, affects the metabolic activation of these mutagens. PMID- 10685009 TI - Inhibitory effects of tea extracts on the mutagenicity of 1-methyl-1, 2,3,4 tetrahydro-beta-carboline-3-carboxylic acid on treatment with nitrite in the presence of ethanol. AB - It has been shown that the mutagenicity of 1-methyl-1,2,3, 4-tetrahydro-beta carboline-3-carboxylic acid (MTCCA), a major mutagen precursor in soy sauce on treatment with nitrite and ethanol, was strongly decreased by the addition of hot water extracts of green, black and oolong teas in the reaction mixture when it was treated with 50mM nitrite at pH3.0, 37 degrees C for 60min in the presence of 7.5% ethanol. The mutagenicity-decreasing activity of the teas was scarcely decreased by washing the teas with chloroform and benzene and was partly decreased by butanol and ethyl acetate. Typical polyphenols such as catechins were shown to have the antimutagenicity dose dependently. The antimutagenicity and the reducing power of tea extracts gave a positive good correlation. The results suggest that the mutagenicity of MTCCA on treatment with nitrite in the presence of ethanol may be decreased by the mixed fractions of lyophilic components such as polyphenols, which have high reducing power such as catechins and the other compounds which have little reducing power including the derivatives of the catechins and so on. Although the antimutagenicity of teas and catechins was also considerably effective when they were added after the nitrosation, that of black tea and some catechins was less effective. PMID- 10685010 TI - Indole-3-carbinol as a chemopreventive agent in 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP) carcinogenesis: inhibition of PhIP-DNA adduct formation, acceleration of PhIP metabolism, and induction of cytochrome P450 in female F344 rats. AB - The chemopreventive properties of dietary indole-3-carbinol (I3C) were evaluated by assessing its effect on DNA adduct formation and metabolism of the dietary carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and the induction of cytochromes P450 1A1 and -1A2 in female F344 rats. In experiment 1, animals on I3C diets (0, 0.02% or 0.1%, w/w) were treated by gavage with 1mg/kg/day of PhIP for 23 days. On days 2, 9, 16 and 23, their 24-hr urine was collected and unmetabolized PhIP was measured by GC/MS. On day 24, the animals were sacrificed, and DNA from pancreas, spleen, white blood cells (WBCs), lung, colon, kidney, mammary epithelial cells, caecum, heart, small intestine, liver and stomach was isolated for determination of PhIP-DNA adduct levels by (32)P postlabelling assays. Except in the mammary gland, I3C diets significantly inhibited PhIP-DNA adduct formation in WBCs and in all organs, ranging from 34.7 to 67.7% with the 0.02% I3C diet to 68.4 to 95.3% with the 0.1% I3C diet. I3C diets also significantly decreased the concentration of urinary unmetabolized PhIP to 29.5-38.4% (0.02% I3C) and 12.8-17.8% (0.1% I3C) of values obtained with the I3C-free diet. In experiment 2, animals were either treated by intubation of I3C at 100 or 200mg/kg for 2 consecutive days or given an I3C-containing diet (0.02% or 0.1%, w/w) for 2 weeks. The expression and activity of cytochromes P450 1A1 and -1A2 were studied by Northern blots, Western blots, and in vitro enzyme determinations. Both the expression and activity of these cytochromes were induced by all of the I3C treatments. It is concluded that, in the female F344 rat, dietary I3C inhibits PhIP-DNA adduct formation and accelerates PhIP metabolism, probably through induction of cytochromes P450 1A1 and -1A2. The chemopreventive properties of I3C in PhIP-induced carcinogenesis are probably mediated through enhancement of PhIP detoxification pathways. PMID- 10685011 TI - Effects of toxaphene on the immune system of cynomolgus (Macaca fascicularis) monkeys. A pilot study. AB - Toxaphene in glycerol/corn oil was administered at 1mg/kg body weight/day, 7 days/week in gelatin capsules to four healthy young adult cynomolgus (Macaca fascicularis) (two male and two female) monkeys for 52 weeks. Control monkeys ingested glycerol/corn oil only. Testing for immune effects was initiated at 34 weeks of treatment. Results included: reduced anti-sheep red blood cell (SRBC) titres for immunoglobulins (Ig) M and G; increased IgG titres to pneumococcal antigens, but not to the tetanus toxoid antigen; reduced T-helper/inducer mean lymphocyte numbers and the mean T-helper/inducer:T-suppressor/cytotoxic cell ratio and reduced respiratory burst activity in peripheral blood monocytes and granulocytes, albeit no changes on the phagocytic activity of these cells were detected. The above noted effects although not statistically significant (P0.05) suggest that chronic exposure to low levels of toxaphene may be immunosuppressive in cynomolgus monkeys and may pose a hazard to human health. To advance our understanding of the degree of hazard that toxaphene may pose to human health, we have undertaken additional chronic studies with a larger number of animals. Particular attention is focused on determining the potential immunotoxic effects of toxaphene in offspring following in utero exposure. PMID- 10685012 TI - A combined subchronic (90-day) toxicity and neurotoxicity study of a single-cell source of docosahexaenoic acid triglyceride (DHASCO oil). AB - Docosahexaenoic acid (DHA), a 22-carbon long-chain polyunsaturated fatty acid of the omega-3 family, is a major structural component of neural membranes and is a particularly important nutrient during infant development. New safe and well defined sources of DHA are required for infant formula fortification and dietary supplementation. DHASCO oil is an algal-derived triglyceride containing 40-50% DHA. Previous studies have shown that DHASCO oil is neither mutagenic nor toxic in acute or 28-day subchronic tests. To further establish the safety of this oil, a 90-day subchronic toxicity study in rats which included haematology, clinical chemistry, pathology and ophthalmologic, neurobehavioural and neuropathological assessments, using doses of 0.5 and 1.25g/kg body weight/day was performed. There were no treatment-related adverse effects in any of the parameters measured at either dose. Based on these results, the no-adverse-effect level (NOAEL) for DHASCO oil under the conditions of this study corresponds to the highest dose level. The DHA in the DHASCO oil was bioavailable, resulting in significant elevations in the levels of this fatty acid in liver, heart and brain after 90 days of administration. In conclusion, this 90-day subchronic toxicity study provides additional evidence that DHASCO oil is a safe and bioavailable source of dietary DHA. PMID- 10685013 TI - Effect of chronic cyanide intoxication on memory in albino rats. AB - Cyanide is a chemical widely used in industry, and is a major environmental pollutant. Its toxicity is caused by inhibition of cytochrome oxidase resulting in histotoxic hypoxia. The effect of sublethal doses of cyanide on memory and hippocampal neurotransmitters was studied in male Wistar strain albino rats. Cyanide reduced the memory along with reduction in the levels of dopamine and 5 hydroxytryptamine in the hippocampus. Pre-existing malnutrition in the animals exaggerated these effects. PMID- 10685014 TI - Developmental toxicity of 2-ethylhexyl stearate. AB - 2-Ethylhexyl stearate was investigated in an embryo-/foetotoxicity and teratogenicity study on rats according to OECD guidelines for the testing of chemicals (No. 414). Dose levels of 0 (arachidis oil), 100, 300 and 1000mg/kg body weight/day were administered by gavage. Dams tolerated the applied dose levels without any toxic effects. Pre- and post-implantation loss and mean numbers of resorptions were unaffected by treatment. All parameters were comparable with the animals of the control group. Skeletal and visceral investigations revealed no treatment-related malformations. For embryo /foetotoxicity, teratogenicity and maternal toxicity a NOAEL of 1000mg/kg was deduced. PMID- 10685015 TI - Clinical, biochemical and neurobehavioural studies of workers engaged in the manufacture of quinalphos. AB - 59 workers exposed to different chemicals during the manufacture of quinalphos, an organophosphate pesticide (OP) and 17 control subjects were studied. Despite similar blood acetylcholinestarase (AChE) levels in both the exposed and control subjects, a significant number of exposed subjects had altered plantar and ankle reflexes. Higher nervous functions such as memory, learning and vigilance were also found to be affected in these subjects. These findings were attributed to chronic low dose combined exposure to different chemicals used/formed in the manufacture of quinalphos. The study raises the doubt that monitoring of AChE alone among subjects engaged in the manufacture of OP pesticides may not be an adequate safeguard as regards to their health. PMID- 10685016 TI - Developmental toxicity study of Aquacoat ECD ethylcellulose aqueous dispersion administered orally to rats. AB - A developmental rat toxicity study of Aquacoat((R)) ECD was performed as part of a program to evaluate the safety of the product. Groups of 25 presumed-pregnant Charles River Sprague-Dawley CD rats received doses of 0, 903, 2709 and 4515mg/kg/day (dry weight basis) of Aquacoat ECD administered undiluted once daily via oral gavage on days 6-15 of gestation. All surviving dams underwent caesarean sectioning on day 20 of gestation. Foetuses were weighed, sacrificed and subject to external, visceral and skeletal evaluations. No test material related maternal deaths occurred; one high-dose female died on day 14 due to gavage error. The only treatment-related clinical sign noted among dams receiving 2709mg/kg/day and greater was pale faeces which was attributed to the presence of the test material in the faeces. No statistically significant differences were noted among the measured maternal parameters. Foetal sex ratios and body weights were similar in all groups. The results of external and visceral foetal evaluations revealed no treatment-related alterations. The only statistically significant findings noted during the skeletal evaluation were increased litter incidences of incompletely ossified or wavy ribs noted among foetuses receiving 4515mg/kg/day, and a significant increase in the litter incidence of thickened ribs at doses of 2709 and 4515mg/kg/day. Given the nature of these findings and the lack of effects on any other parameter measured in this study, they were not considered adverse effects of treatment. Under the conditions of this study, the maternal and foetal no-observed-adverse-effect level (NOAEL) is in excess of 4515mg/mg/day. PMID- 10685017 TI - In vivo cytogenetic studies on blends of aspartame and acesulfame-K. AB - Aspartame and acesulfame-K, non-nutritive sweeteners, are permitted individually in diets and beverages. These sweeteners of different classes, used in combination, have been found to possess a synergistic sweetening effect. Whether they also have a synergistic genotoxic effect is unknown. Swiss Albino male mice were exposed to blends of aspartame (3.5, 35, 350mg/kg body weight) and acesulfame-K (1.5, 15 and 150mg/kg body weight) by gavage. Bone marrow cells isolated from femora were analysed for chromosome aberrations. Statistical analysis of the results show that aspartame in combination with acesulfame-K is not significantly genotoxic. PMID- 10685018 TI - Review of the toxicologic properties of medium-chain triglycerides. AB - Medium chain triglycerides (MCTs) are a family of triglycerides, containing predominantly, caprylic (C(8)) and capric (C(10)) fatty acids with lesser amounts of caproic (C(6)) and lauric (C(12)) fatty acids. MCTs are widely used for parenteral nutrition in individuals requiring supplemental nutrition and are being more widely used in foods, drugs and cosmetics. MCTs are essentially non toxic in acute toxicity tests conducted in several species of animals. In ocular and dermal irritation testing MCTs exhibit virtually no potential as ocular or dermal irritants, even with prolonged eye or skin exposure. MCTs exhibit no capacity for induction of hypersensitivity. Ninety-day toxicity tests did not result in notable toxicity, whether the product was administered in the diet up to 9375mg/kg body weight/day or by intramuscular (im) injection (up to 0. 5ml/kg/day, rabbits). There was no evidence that intravenous (iv) or dietary administration of MCTs adversely affected the reproductive performance of rats or resulted in maternal toxicity, foetal toxicity or teratogenic effects at doses up to 4.28g/kg body weight/day (iv) or 12,500mg/kg body weight/day (dietary). There was no evidence that dietary administration of MCTs adversely affected the reproductive performance of pigs or resulted in maternal toxicity, foetal toxicity or teratogenic effects at doses up to 4000mg/kg body weight/day in the diet. In rabbits, following iv administration, the maternal and foetal no observed-adverse-effect levels (NOAELs) were between 1.0 and 4.28g/kg body weight/ day. A 2-year study in rats, conducted with a closely related compound (tricaprylin, a triglyceride with C(8) fatty acids), provided no evidence of a carcinogenic effect when the material was administered by oral gavage at levels up to 10ml/kg (9.54g/kg) per day. Although tricaprylin was found to be positive in one of five strains of Salmonella typhimurium in the presence of metabolic activation in an Ames mutagenicity assay, the results of the carcinogenicity test with tricaprylin and mutagenicity tests with caprylic acid indicate that MCTs do not have the potential to be carcinogenic or mutagenic. The safety of human dietary consumption of MCTs, up to levels of 1g/kg, has been confirmed in several clinical trials. PMID- 10685019 TI - Safety assessment of iron EDTA [sodium iron (Fe(3+)) ethylenediaminetetraacetic acid]: summary of toxicological, fortification and exposure data. AB - Iron EDTA [sodium iron (Fe(3+)) ethylenediaminetetraacetic acid (EDTA)], shown to have a significant beneficial effect on iron status by increasing iron bioavailability in human diets, has been proposed for use as a fortificant in certain grain-based products including breakfast cereals and cereal bars. This paper presents an assessment of the safety of iron EDTA for its intended uses in these products. Iron EDTA, like other EDTA-metal complexes, dissociates in the gastrointestinal tract to form iron, which is bioavailable, and an EDTA salt; absorption of the metal ion and EDTA are independent. Because of this dissociation, consideration of information on EDTA compounds other than iron EDTA is relevant to this safety assessment. EDTA compounds are poorly absorbed in the gastrointestinal tract and do not undergo significant metabolic conversion. They have a low degree of acute oral toxicity. EDTA compounds are not reproductive or developmental toxicants when fed with a nutrient-sufficient diet or minimal diets supplemented with zinc. In chronic toxicity studies, diets containing as much as 5% EDTA were without adverse effects. EDTA compounds were not carcinogenic in experimental animal bioassays and are not directly genotoxic. This lack of significant toxicity is consistent with a history of safe use of other EDTA compounds (CaNa(2)EDTA and Na(2)EDTA) approved by the FDA for use as direct food additives. An upper-bound estimated daily intake (EDI) of EDTA from iron EDTA (1.15mg/kg bw/day for the US population) is less than half the acceptable daily intake (ADI) for EDTA of 2. 5mg/kg bw/day established by JECFA. The data collected and published over the past 20 to 30 years demonstrate that iron EDTA is safe and effective for iron fortification of food products and meets the standard of "reasonable certainty of no harm". Based on the published record, iron EDTA may be regarded as generally recognized as safe (GRAS) for the intended food uses and maximum use levels. PMID- 10685020 TI - The 'considerate' smoker in public space: the micro-politics and political economy of 'doing the right thing'. AB - This paper examines the discourse of 'interactions' as applied to the interpersonal management of smoking in public places (and to accounts thereof). Empirical data from a qualitative study of smokers and non-smokers in metropolitan Toronto, Ontario (Canada) are used to illustrate how smokers and non smokers define and claim to operationalize 'consideration' in their daily lives. Drawing on the work of Foucault, Rose, Castel, and Bourdieu, the paper explores the possible significance of 'consideration' as a discourse of risk management masked as 'common sense', as a marker of social competence. In particular, parallels with emergent forms of governmentality embedded in community participation and individual self-monitoring and self-restraint are noted. Further, the social control implications of 'consideration' as moral discourse are examined with respect to Bourdieu's analysis of class struggles for (social) distinction. In this light, it is suggested that legitimate health concerns raised by tobacco control advocates cannot be divorced from other implicit social agendas which also fuel the drive for the 'purification of public space'. PMID- 10685021 TI - Efficiency of care at the primary-secondary interface: variations with GP fundholding. AB - The aim of the study was to investigate the processes of referral for out patients care and the interface with general practice, from the perspective of the patient, the patient's general practitioner and hospital specialist. The analyses reported here present variations with fundholding and non-fundholding general practice. The design was a questionnaire survey of out-patients, their hospital specialists and general practitioners, in six, randomly sampled district health authorities in the North Thames Region, with stratification by area. The measures included validated items and scales on process, quality and patient satisfaction with services. Fundholders were more likely to have technical equipment and services available within the practice. There were no differences between fundholders and non-fundholders and the number of out-patient attendances made by their patients, hospital out-patient waiting list times, patients' waiting times in hospital clinics, nor in patients' satisfaction with out patients and other process indicators. Fundholding is currently being replaced with the proposed wider locality commissioning schemes, with GPs, health authorities and other purchasing bodies acting in partnership. Health authority commissioning will be required to reflect the preferences of GPs. Participants in these schemes will need to pay particular attention to the areas where research indicates that fundholding GPs made little difference to increasing the efficiency and effectiveness of health care both in their own practices and at the primary-secondary care interface. PMID- 10685022 TI - Health and hygiene knowledge, attitudes and behaviour. AB - The aim of this study was to develop and test measures of health and hygiene knowledge, attitudes and behaviour. A questionnaire was administered to 240 women: 80 from a squatter camp, 80 from an informal settlement and 80 from a formal township. Reliability of the knowledge scale was 0.73. Coefficient alpha was 0.87 for the attitude and behaviour scales. The knowledge, attitude and behaviour scales were significantly related (P<0.001). Factor analysis confirmed that domestic and personal hygiene were core components of the attitude scale, whereas the emphasis for behaviour was on personal hygiene. Stepwise regression showed that age explained 23% of the variance in knowledge and 18% in behaviour. Waste management significantly affected knowledge, attitudes and behaviour, suggesting that dustbin ownership was an important public health measure. It was concluded that these scales were useful measures of health and hygiene knowledge, attitudes and behaviour; provided baseline information for planning health promotion programmes; and could be used to evaluate the effectiveness of such programmes. PMID- 10685023 TI - Determinants of geographic mobility among participants in a population-based HIV/AIDS drug treatment program. AB - This study was undertaken to determine the geographic distribution and patterns of migration of persons with HIV in British Columbia. Our analysis was restricted to all HIV-positive men and women aged 18 years and over who had completed a participant survey and were enrolled in the HIV/AIDS Drug Treatment Program between September 1992 and September 1997. Patterns of migration were determined by examining participants whose postal code changed between July 1995 and September 1997. Statistical analysis were carried out using both parametric and non-parametric methodologies. Stepwise logistic regression was used to determine baseline predictors of migration. The final multivariate model revealed that residing in a census subdivision with a population less than 100,000, being heterosexual, acquiring HIV through intravenous drug use, and the absence of AIDS at baseline were all independently associated with moving census subdivisions during the period of observation. In summary, our analyses demonstrate the need for the continued study of the evolving geography and migration patterns of persons with HIV. PMID- 10685024 TI - Urbanization and the urban mortality change in Imperial Germany. AB - This analysis assesses urban mortality change in Imperial Germany, when the country was going through a process of accelerated industrialization and urbanization. Urban mortality reached its peak after the middle of the century, thereafter urban mortality improved substantially. The largest cities, particularly in the highly industrialized western parts of the country, registered the strongest decline. A key element in this process was the reduction of mortality from gastrointestinal disorders, affecting almost exclusively infants. Therefore this analysis discusses the impact of selected public health strategies designed to fight high infant mortality rates. Special emphasis is placed on municipal milk supply and infant welfare centres. PMID- 10685025 TI - Health, place and British prisons. AB - This short communication considers people's health care needs in prison. It shows that people have a wide range of health care needs that are not always met when they spend time in prison. To locate these issues the paper draws on the "patient or prisoner" debate to understand the challenges that face future policies. PMID- 10685026 TI - Biochemical and compositional analyses of recombinant lecithin:cholesterol acyltransferase (LCAT) obtained from a hepatic source. AB - Lecithin:cholesterol acyltransferase (LCAT) is an important plasma glycoprotein which plays a central role in lipid metabolism. This protein is responsible for generation of cholesteryl esters in plasma and it has been proposed to play a pivotal role in the reverse cholesterol transport pathway. Structural and functional studies of LCAT have employed various expression systems for production of recombinant LCAT (rLCAT). However, recent studies have shown some differences in the oligosaccharide structure and composition of rLCAT. In this study, we have generated a new hepatic based expression system using McArdle RH7777 (Mc-7777) cells to produce a recombinant protein most similar to human plasma LCAT. The expressed glycoprotein was compared to the LCAT expressed in previously characterized baby hamster kidney (BHK) cells. Both proteins were compared on the basis of their carbohydrate structure and composition as well as their functional properties. Although the functional properties of both glycoproteins were similar, the carbohydrate structure was significantly different. While BHK-LCAT contained bi-, tri-, and tetraantennary structures, Mc 7777 LCAT presented only biantennary oligosaccharide structures. The difference in glycosylation pattern of rLCAT from Mc-7777 and BHK cells underlines the importance of appropriate expression system, both in vivo and in vitro. PMID- 10685027 TI - Conformation of apolipoprotein E both in free and in lipid-bound form may determine the avidity of triglyceride-rich lipoproteins to the LDL receptor: structural and kinetic study. AB - Slow refolding of human apolipoprotein E (apoE) in solution after guanidine- or cholate-induced denaturation followed by dialysis under controlled conditions was investigated using various spectroscopic properties of fluorescein- and dansyl labeled apolipoprotein molecules. The results suggest that the last phase(s) of apoE refolding in solution include a slow (several hours at 24 degrees C) interconversion of a self-associated 'open' conformer into a more dense 'closed' conformer. The hydrophobic interactions are primarily responsible for the formation of this more compact apoE structure. To visualize the contribution of apolipoprotein conformation and/or the number of 'active' lipid-bound apoE molecules in the reaction of binding to the low density lipoprotein receptor (LDLr) by solid-phase binding assay, the complexes of human plasma apolipoprotein or recombinant (rec) apoE3 with dipalmitoylphosphatidylcholine (DPPC) or palmitoyloleoylphosphatidylcholine (POPC) varying in size were used. For seven complexes with plasma protein (four DPPC and three POPC complexes), the final phosphatidylcholine (PC)/protein mole ratio ranged from 117 to 279; affinity constant K(a) averaged for both PCs and plotted against this ratio abruptly increased from 3.8 x 10(7) to 3.8 x 10(8) M(-1) with a transition midpoint of 150 180 PC/apoE, mole ratio. Two DPPC complexes with rec protein bind much more efficiently. Complexes with both plasma and rec apoE were able to compete with very low density lipoproteins (VLDL) or low density lipoproteins (LDL) isolated from patients with E3/3 phenotype, for binding to the LDLr. Again, the competition efficiency abruptly increased at the increase in PC content with a transition midpoint of 130 PC/apoE, mole ratio. The transitions observed both in direct and competitive binding assay probably correspond to the abrupt increase in the number of 'active' apoE molecules on the complex surface accompanying the change in the size and/or in the shape of the complexes. The efficiency of apoE and apoB as the corresponding major ligands in the binding reaction of VLDL and LDL to the LDL receptor was compared. VLDL bind to LDLr following a simple encounter complex model, while LDL binding was characterized by a more complex two-step model with an additional isomerization step. The analysis of the binding data led us to suggest the existence of the continuum from several (2-3) apoE molecules on the surface of TG-rich particles that resulted in the increased binding affinity, on average 3.5-fold higher, compared to LDL. The existence of a complex equilibrium between aqueous and different lipid-bound forms of apoE is proposed, in particular, the formation of a transient disc-lipoprotein particle structure during the interaction with LDLr in vivo as well as in LPL-stimulated lipolysis of the lipid phase of the particle. PMID- 10685028 TI - Structural peculiarities of the binding of very low density lipoproteins and low density lipoproteins to the LDL receptor in hypertriglyceridemia: role of apolipoprotein E. AB - Very low (VLDL) and low density lipoproteins (LDL) were isolated from plasma of patients with the E3/3 phenotype which were divided into three groups based on their plasma triglyceride content: low (TG<200 mg/dl, TG(l)), intermediate (200<300 mg/dl, TG(i)300 mg/dl, TG(h)). The protein density (PD) on the VLDL and LDL surface was calculated from lipoprotein composition and protein location was studied by tryptophan fluorescence quenching by I(-) anions at 25 degrees C and 40 degrees C. A comparison of the TG(h) with the TG(l) group revealed a significant (<0.05) increase of the PD parameter as much as 21% for VLDL, but not for LDL where this parameter did not change for any group; generally, PD(LDL) values were 3.2-3.8-fold lower than PD(VLDL). In accordance with this difference, the tryptophan accessibility f in VLDL vs. LDL was lower at both temperatures. There were temperature-induced changes of the f parameter in opposite directions for these lipoproteins. The difference in f value gradually decreased for VLDL in the direction TG(l)TG(i)TG(h) while for LDL there was a U-shaped dependence for these groups. The Stern-Volmer quenching constant K(S-V) which is sensitive to both temperature and viscosity, did not change for VLDL, but K(S-V)(LDL) was 2-3 fold higher for the TG(i) group compared to the other two. The efficiencies of VLDL and LDL binding to the LDL receptor (LDLr) in vitro were compared by solid phase assay free of steric hindrance observed in cell binding. The maximal number of binding sites did not change for either type of particles and between groups. The association constant K(a) and apolipoprotein (apo) E/apoB mole ratio values all increased significantly for VLDL, but not for LDL, in comparison of the TG(i+h) with the TG(l) group. Based on VLDL and LDL concentrations in serum and on the affinity constant values obtained in an in vitro assay, VLDL concentrations corresponding to 50% inhibition of LDL binding (IC(50)) were calculated in an assumption of the competition of both ligands for LDLr in vivo; the mean values of IC(50) decreased 2-fold when plasma TG exceeded 200 mg/dl. The functional dependences of K(a)(VLDL), IC(50) and apoE content in VLDL (both fractional and absolute) and in serum on TG content in the whole concentration range studied were fitted to a saturation model. For all five parameters, the mean half-maximum values TG(1/2) were in the range 52-103 mg/dl. The efficiency of protein-protein interactions is suggested to differ in normolipidemic vs. HTG VLDL and apoE content and/or protein density on VLDL surface may be the primary determinant(s) of the increased binding of HTG-VLDL to the LDL receptor. ApoCs may compete with apoE for the binding to the VLDL lipid surface as plasma triglyceride content increases. The possible competition of VLDL with LDL for the catabolism site(s) in vivo, when plasma TG increases, could explain the atherogenic action of TG-rich lipoproteins. Moreover, the 'dual action' hypothesis on anti-atherogenic action of apoE-containing high density lipoproteins (HDL) in vivo is suggested: besides the well-known effect of HDL as cholesteryl ester catabolic outway, the formation of a transient complex of apoE containing discs appearing at the site of VLDL TG hydrolysis by lipoprotein lipase with VLDL particles proposed in our preceding paper promotes the efficient uptake of TG-rich particles; in hypertriglyceridemia due to the diminished HDL content this uptake seems to be impaired which results in the increased accumulation of the remnants of TG-rich particles. This explains the observed increase in cholesterol and triglyceride content in VLDL and LDL, respectively, due to the CETP-mediated exchange of cholesteryl ester and triglyceride molecules between these particles. PMID- 10685029 TI - Upregulation of uncoupling protein 2 mRNA in genetic obesity: lack of an essential role for leptin, hyperphagia, increased tissue lipid content, and TNF alpha. AB - Uncoupling protein 2 (UCP2) has been proposed to play a prominent role in the regulation of energy balance. UCP2 mRNA expression is upregulated in white adipose tissue (WAT) and liver, but is not altered in skeletal muscle in genetically obese ob/ob mice. The mechanisms involved in the upregulation of UCP2 in obesity have not been investigated. We have now examined the potential role of leptin, hyperphagia, increased tissue lipid content, and overexpression of tumor necrosis factor (TNF)-alpha in the upregulation of UCP2 mRNA expression in the liver and WAT in ob/ob mice. Treatment of ob/ob mice with leptin for 3 days significantly reduced their food intake but had no effect on the upregulation of UCP2 mRNA levels in the liver or WAT. To investigate the effect of feeding and higher tissue lipid content on the upregulation of UCP2 in liver and WAT, we compared UCP2 mRNA levels in ad-libitum fed and 72-h fasted control and ob/ob mice. In controls, fasting had no effect on UCP2 mRNA levels in liver, but increased UCP2 mRNA in WAT suggesting that the effects of fasting on UCP2 mRNA levels are tissue-specific. In ob/ob mice, fasting did not lower UCP2 mRNA levels in liver or WAT suggesting that the upregulation of UCP2 in ob/ob mice is not merely a direct consequence of increased food intake. 72-h fasting lowered hepatic total lipid content by 34% and 36% in control and ob/ob mice, respectively, without any corresponding decrease in hepatic UCP2 mRNA levels, suggesting that the enhanced UCP2 expression in the liver of ob/ob mice is not secondary to lipid accumulation in their livers. Although TNF-alpha has been shown to acutely increase UCP2 mRNA levels in liver and WAT, and is overexpressed in adipose tissue in obesity, deletion of the genes for both TNF receptors in ob/ob mice produces a further increase in UCP2 mRNA expression in liver and adipose tissue indicating a paradoxical inhibitory role. Taken together, these results suggest that the upregulation of UCP2 mRNA levels in the liver and WAT of ob/ob mice is not due to the lack of leptin, hyperphagia, increased tissue lipid content, or over-expression of TNF-alpha. PMID- 10685030 TI - LTA(4)-derived 5-oxo-eicosatetraenoic acid: pH-dependent formation and interaction with the LTB(4) receptor of human polymorphonuclear leukocytes. AB - 5-oxo-(7E,9E,11Z,14Z)-eicosatetraenoic acid (5-oxo-ETE) has been identified as a non-enzymatic hydrolysis product of leukotriene A(4) (LTA(4)) in addition to 5,12 dihydroxy-(6E,8E,10E, 14Z)-eicosatetraenoic acids (5,12-diHETEs) and 5,6 dihydroxy-(7E,9E, 11Z,14Z)-eicosatetraenoic acids (5,6-diHETEs). The amount of 5 oxo-ETE detected in the mixture of the hydrolysis products of LTA(4) was found to be pH-dependent. After incubation of LTA(4) in aqueous medium, the ratio of 5-oxo ETE to 5,12-diHETE was 1:6 at pH 7.5, and 1:1 at pH 9.5. 5-Oxo-ETE was isolated from the alkaline hydrolysis products of LTA(4) in order to evaluate its effects on human polymorphonuclear (PMN) leukocytes. 5-Oxo-ETE induced a rapid and dose dependent mobilization of calcium in PMN leukocytes with an EC(50) of 250 nM, as compared to values of 3.5 nM for leukotriene B(4) (LTB(4)500 nM for 5(S)-hydroxy (6E,8Z,11Z,14Z)-eicosatetraenoic acid (5-HETE). Pretreatment of the cells with LTB(4) totally abolished the calcium response induced by 5-oxo-ETE. In contrast, the preincubation with 5-oxo-ETE did not affect the calcium mobilization induced by LTB(4). The calcium response induced by 5-oxo-ETE was totally inhibited by the specific LTB(4) receptor antagonist LY223982. These data demonstrate that 5-oxo ETE can induce calcium mobilization in PMN leukocyte via the LTB(4) receptor in contrast to the closely related analog 5-oxo-(6E,8Z,11Z, 14Z)-eicosatetraenoic acid which is known to activate human neutrophils by a mechanism independent of the receptor for LTB(4). PMID- 10685031 TI - Analysis of sulfatide from rat cerebellum and multiple sclerosis white matter by negative ion electrospray mass spectrometry. AB - The accumulation of sulfatide (sulfatogalactosyl cerebroside) and changes in the sulfatide species present have been examined in the cerebellum of day 6-32 aged rats and in multiple sclerosis (MS) tissue samples. Negative ion electrospray mass spectrometry with daughter and parent ion analyses were used to distinguish the fatty acyl character in the amide linkage of sulfatide; measurement was done by selected ion and multiple reaction monitoring of individually identified sulfatide molecules. Sulfatide accumulation in rat cerebellum shows that 18:0- and hydroxylated 18:0-sulfatide are the first sulfatide molecules detectable. Very long fatty acyl chain sulfatide molecules (>20:0) are present at day 7 and the ratio of non-hydroxylated compared to hydroxylated sulfatide rises as the amount of non-hydroxylated sulfatide increases. 24:1-sulfatide accumulates at a ratio of about 3:1 over 24:0-sulfatide during active myelination. Analyses of the sulfatide in human tissue have shown differences between MS plaque tissues, normal appearing adjacent white matter and control tissues. The findings show that total sulfatide is reduced by 60% in the plaque matter and decreased 25% in adjacent normal appearing white matter. There are significant increases (P=0.05) in the amount of hydroxylation of sulfatide, demonstrated by an increase in the percentage of hydroxylated h24:0-sulfatide (hydroxy-lignoceroyl sulfatide). PMID- 10685032 TI - Enzymological properties of the LPP1-encoded lipid phosphatase from Saccharomyces cerevisiae. AB - The product of the LPP1 gene in Saccharomyces cerevisiae is a membrane-associated enzyme that catalyzes the Mg(2+)-independent dephosphorylation of phosphatidate (PA), diacylglycerol pyrophosphate (DGPP), and lysophosphatidate (LPA). The LPP1 encoded lipid phosphatase was overexpressed 681-fold in Sf-9 insect cells and used to examine the enzymological properties of the enzyme using PA, DGPP, and LPA as substrates. The optimum pH values for PA phosphatase, DGPP phosphatase, and LPA phosphatase activities were 7. 5, 7.0, and 7.0, respectively. Divalent cations (Mn(2+), Co(2+), and Ca(2+)), NaF, heavy metals, propranolol, phenylglyoxal, and N-ethylmaleimide inhibited the PA phosphatase, DGPP phosphatase, and LPA phosphatase activities of the enzyme. The inhibitory effects of N-ethylmaleimide and phenylglyoxal on the LPP1-encoded enzyme were novel properties when compared with other Mg(2+)-independent lipid phosphate phosphatases from S. cerevisiae and mammalian cells. The LPP1-encoded enzyme exhibited saturation kinetics with respect to the surface concentrations of PA (K(m)=0.05 mol%), DGPP (K(m)=0.07 mol%), and LPA (K(m)=0.08 mol%). Based on specificity constants (V(max)/K(m)LPA (1.3 units/mg/mol%). DGPP (K(i)=0.12 mol%) was a competitive inhibitor with respect to PA, and PA (K(i)=0.12 mol%) was a competitive inhibitor with respect to DGPP. This suggested that the binding sites for these substrates were the same. The enzymological properties of the LPP1 encoded enzyme differed significantly from those of the S. cerevisiae DPP1 encoded lipid phosphatase, a related enzyme that also utilizes PA, DGPP, and LPA as substrates. PMID- 10685033 TI - A view on the science: physical anthropology at the millennium. PMID- 10685034 TI - Maxillary sinusitis as an indicator of respiratory health in past populations. AB - Chronic infectious respiratory disease in a past human population is investigated through the quantification of maxillary sinusitis among Iroquoian horticulturists. Three hundred forty-eight right and left maxillae of a Southern Ontario Iroquoian skeletal sample, Uxbridge Ossuary, ca. AD 1440, were examined for evidence of chronic infection (minimum number of individuals = 207: 114 adults, 22 adolescents, 38 juveniles and 33 infants). Modern clinical criteria were applied to differentiate lesions of respiratory and dental origin. Osseous lesions of the maxillary sinuses were observed in 50% of the individuals examined. These lesions are morphologically consistent with nonspecific lesions observed in other past populations that have been attributed to the presence of pathogens. The prevalence of maxillary sinusitis increases with age. Osseous changes suggestive of maxillary sinusitis of respiratory origin are at a maximum prevalence in juveniles and adolescents. In adults, infection of dental origin becomes a confounding factor in the identification of sinusitis of respiratory origin. Fifteenth century Iroquoians were experiencing high airborne pathogen levels and poor indoor air quality. The prevalence of maxillary sinusitis and the exploration of the origin of tissue injury may contribute to our reconstruction of the quality of life and the respiratory health status of past human populations. PMID- 10685035 TI - Cribra orbitalia in two temporally disjunct population samples from the Dakhleh Oasis, Egypt. AB - Cribra orbitalia (CO), an osseous sign of anemic stress, occurs in 67% (n = 296) of the pre-Roman (n = 153) and Roman (n = 143) period crania from the Dakhleh Oasis, Egypt. CO is primarily a childhood condition in these samples, and its prevalence is significantly higher in virtually all cohorts in the pre-Roman group, including among females, who display higher rates of active lesions. This temporal trend suggests that the underlying causative factors (i.e., synergism between disease and nutrition) were less pervasive in the Roman period. In both population samples, anemic stress develops in some perinates prior to the expected minimum age for the development of iron deficiency anemia. This suggests additional causes of anemic stress in the Dakhleh population. A strong candidate is folic acid deficiency and its concomitant, megaloblastic anemia, which results from weaning of infants on goat's milk, a known practice in ancient Egypt. The putative incorporation of other food items in the weanling diet, particularly honey, a confirmed source of C. botulinum, represents yet another retrospective data source to help understand the epidemiological profile of cribra orbitalia in this population. Comparative data from other Egyptian populations, though limited, show similar patterns, however, they display a lower prevalence than the data from Dakhleh. PMID- 10685036 TI - Sex differences in activity-related osseous change in the spine and the gendered division of labor at Ensay and Wharram Percy, UK. AB - Sex differences in the distribution of vertebral degenerative and plastic change were examined and compared within and between samples of 51 individuals from the historically and ethnographically documented 16th-19th century site of Ensay, the Outer Hebrides, and 59 individuals from the medieval site of Wharram Percy, the Yorkshire Wolds. Both populations have a known gendered division of labor between males and females and known activity-related stresses on the spine. Osseous changes normally associated with degenerative joint disease (osteoarthritis) of the apophyseal facets and osteophytosis of the vertebral bodies were scored and reported separately. Inter- and intrasite differences were found in the frequency and distribution of osseous change down the spine. Overall, the Ensay sample was more highly stressed than that from Wharram Percy. Furthermore, differences between males and females at Ensay could be identified as relating to different types of activities. Distinctions between males and females at Wharram Percy were less marked, suggesting broadly similar lifestyles. These results accorded with expectations regarding contrasting levels of activity-related stress at the two sites and the division of labor between males and females. In particular, the prevalence and distribution of facet remodeling, facet sclerosis/eburnation, and osteophytosis in Ensay females could be related to load-bearing using creels (a form of basket), which disrupted "normal" patterns of osseous change along the spine. Importantly, morphologically distinct osseous modifications recorded on the apophyseal facets produced dissimilar distributions, suggesting that they may have different etiologies. These results highlight the need for a high degree of discrimination in recording, analyzing, and exploring activity-related osseous change. PMID- 10685037 TI - A sequential developmental field defect of the vertebrae, ribs, and sternum, in a young woman of the 12th century AD. AB - Changes in the vertebral column are often noted in skeletal material. Descriptions of these anomalies are often lacking, and their developmental origins are not often discussed. The skeleton of a young woman from the medieval cemetery of Tirup, in Denmark, has multiple defects of the axial skeleton, including extra thoracic and lumbar vertebrae, border shifting, extra ribs, block vertebra, and deformed sternum. This case study is particularly interesting because of the number and diversity of anomalies seen; the rarity of these defects, even in living populations; and her survival to adult age. Careful analysis of the bones and use of the morphogenetic method of determining development stages has led to the conclusion that the initial defect probably occurred very early in development, during blastogenesis, with the initial development of at least two extra somitomeres in the paraxial mesoderm. These extra elements in turn led to problems in union and differentiation, and later chondrification and ossification of the vertebra. The malformations of the vertebrae also induced changes in the ribs and sternum. The initial error of segmentation is identified as a developmental field defect, and the cascade of anomalies seen is a developmental sequence caused by the initial field defect. The genetic and environmental causes of developmental field defects are reviewed. PMID- 10685038 TI - Tooth wear and compensatory modification of the anterior dentoalveolar complex in humans. AB - In populations living in environments where teeth wear severely, some compensatory modification of the dentoalveolar complex is thought to occur during life whereby functional occlusion is maintained as tooth substance is lost by wear. This study investigates one aspect of this modification process: Changes in the anterior dentoalveolar complex that are accompanied with wear were examined in a series of Japanese skeletal samples. In the prehistoric Japanese hunter gatherer population heavy wear occurs over the entire dentition. The following changes were demonstrated to have occurred in the anterior segment of the dentition accompanied by wear on the anterior teeth: The anterior teeth tip lingually with wear up to a nearly upright position to fill in interproximal spaces that would have been generated by wear, and to maintain contact relations between adjacent teeth. At the same time, the anterior surface of the maxillary alveolar process also inclines lingually to a certain extent. The amount of lingual tipping is greater in the maxillary anterior teeth than in their mandibular antagonists. It is because of this discrepancy that, with age, the horizontal component of the overlap between maxillary and mandibular anterior teeth decreases, and their bite form changes from scissor bite to edge-to-edge bite. Lesser degrees of lingual tipping of the anterior teeth were also detected in the prehistoric agriculturists and historic Japanese populations. The variation in the degree of lingual tipping observed among the samples is explained by inter-population variation in severity and pattern of tooth wear. This and other evidence suggests that mechanisms that compensate for wear in the anterior dentition may be characteristic of all living human populations, independently of the degree of wear severity endured in their environments. PMID- 10685039 TI - Krapina 1: a juvenile Neandertal from the early late Pleistocene of Croatia. AB - The juvenile A Skull from Krapina, Croatia (Krapina 1) has been the subject of considerable debate since B. Skerlj first suggested that it might not be a Neandertal. Although widely known by its original designation, the Krapina A Skull was recatalogued, along with all of the Krapina hominids, in the 1980's (Radovcic, et al., [1988]. The Krapina Hominids: An Illustrated Catalog of Skeletal Collection. Zagreb; Mladost). It is now catalogued as Krapina 1 in the archives of the Hrvatski Prirodoslovni Muzej, Zagreb, Croatia. We present a detailed, morphometric analysis of this specimen, comparing it to other Krapina specimens, juvenile late Pleistocene hominids (including Neandertals), and subadult recent humans. This analysis demonstrates that Krapina 1 possesses morphological features that are primitive retentions; others that represent derived Neandertal specializations; and still others that are typical for all European late Pleistocene humans. Morphological features associated with the browridges are intermediate between Neandertal and early modern European form. Nevertheless, a thorough analysis of the morphology of this specimen, in ontogenetic and regional contexts, leads to the conclusion that it cannot be excluded from the Neandertal range of variation. We conclude that the most parsimonious explanation for this 130 ka specimen is that it should be regarded as a Neandertal. PMID- 10685040 TI - Technical note: Stafne static mandibular bone defect-further expression on the buccal aspect of the ramus. AB - The anatomic distribution of the Stafne static mandibular bone defect (SSBD) is extended with a description of a cavitation defect on the buccal ramus of the mandible. The anatomical placement of SSBD thus correlates precisely with the submandibular and parotid salivary glands, and gives further evidence that an increase in major salivary gland size is associated with the defect. The global latitudinal variation in the population prevalence of SSBD ranges from 10% in the tropics to virtually 0% in most of the arctic. Globally the defect directly correlates with parasite load and diversity, and may be a marker for of a history of an environment with high levels of enteric macroparasite infestation. PMID- 10685041 TI - Brief communication: a study of the predictive accuracy of mandibular ramus flexure as a singular morphologic indicator of sex in an archaeological sample. AB - Loth and Henneberg ([1996] Am J Phys Anthropol 99:473-485) assert that they have discovered a single morphologic indicator of sexual dimorphism in the human mandible that rivals the predictive accuracy of the complete pelvis at 94.2% for all samples (99% for healthy samples). To test the accuracy of their method, mandibles (n = 150) from the Tepe Hissar collection were assessed for the presence or absence of mandibular ramus flexure. These results were then compared to a separate sex assessment based on morphologic indicators from the corresponding skull and innominates (where possible) to yield an overall accuracy of only 78.2%. As a means of independent assessment, the mandibular results were also compared with Krogman's ([1940] Racial Types from Tepe Hessar, Iran, from late fifth to early second millennium, BC. Amsterdam: Koninkliijke Nederlandsche Akademie van Wetenschappen) assessment of sex based on craniofacial measurements and morphologic indicators from the skull. This comparison produced an even lower accuracy of 67.2%. Such results question the predictive potential of mandibular ramus flexure as a single indicator of sexual dimorphism and suggest caution when applying this method, especially in the case of fragmentary forensic and fossil remains. PMID- 10685042 TI - Ligation and splicing of peptides and proteins. PMID- 10685043 TI - Orthogonal ligation strategies for peptide and protein. AB - This review focuses on the concept, criteria, and methods of an orthogonal amide ligating strategy suitable for syntheses of peptides, peptide mimetics, and proteins. Utilizing unprotected peptides or proteins derived from chemical or biosynthetic sources, this ligation strategy has been shown to be general and exceptionally mild. Its orthogonality in ligating two unprotected segments with free N-terminal (NT)-amines at a specific NT-amine is achieved through a chemoselective capture step and then an intramolecular acyl transfer reaction. Both coupling reagents for enthalpic activation and protection schemes therefore become unnecessary. More than a dozen orthogonal ligation methods based on either imine or thioester captures have been developed to afford native and unusual amino acids at ligation sites of linear, branched, or cyclic peptides. Because unprotected peptides and proteins of different sizes and forms can be obtained from either chemical or recombinant sources, orthogonal ligation removes the size limitation imposed on the chemical synthesis of a protein with a native or non native structure. Furthermore, by using building blocks from biosynthetic sources, orthogonal ligation provides a unifying operational concept for both total and semisynthesis of peptides and proteins. PMID- 10685044 TI - Intein-mediated protein ligation: harnessing nature's escape artists. AB - Inteins are naturally occurring proteins that are involved in the precise cleavage and formation of peptide bonds in a process known as protein splicing. Genetic engineering has allowed the controllable cleavage of peptide bonds at either the N- or C-terminus of the intein. Inteins displaying controllable cleavage have been used in the isolation of bacterially expressed proteins possessing either a C-terminal thioester or an N-terminal cysteine. The specific placement of these reactive groups has allowed either protein-protein or protein peptide condensation through a native peptide bond. This review describes the methods used to specifically generate these reactive groups on bacterially expressed proteins and some applications of this technique, known as intein mediated protein ligation. Furthermore, a versatile two intein (TWIN) system will be described which enables the circularization and polymerization of bacterially expressed proteins or peptides. PMID- 10685045 TI - Introduction of unnatural amino acids into proteins using expressed protein ligation. AB - Here we describe the results of studies designed to explore the scope and limitations of expressed protein ligation (EPL), a protein semisynthesis approach that allows unnatural amino acids to be site specifically introduced into large proteins. Using Src homology 3 domains from the proteins c-Abl and c-Crk as model systems, we show here that EPL can be performed in the presence of moderate concentrations of the chemical denaturant, guanidine hydrochloride, and the organic solvent dimethylsulfoxide. Use of these solubilizing agents allowed the successful preparation of two semisynthetic proteins, 10 and 12, both of which could not be prepared using standard procedures due to the low solubility of the synthetic peptide reactants in aqueous buffers. We also report the results of thiolysis and kinetic studies which indicate that stable alkyl thioester derivatives of recombinant proteins can be generated for storage and purification purposes, and that 2-mercaptoethanesulfonic acid compares favorably with thiophenol as the thiol cofactor for EPL reactions, while having superior handling properties. Finally, we describe the semisynthesis of the fluorescein/rhodamine-containing construct (12) and the ketone-containing construct (14). The efficiency of these two syntheses indicates that EPL offers a facile way of incorporating these important types of biophysical and biochemical probes into proteins. PMID- 10685046 TI - Characteristics of protein splicing in trans mediated by a semisynthetic split intein. AB - Protein splicing in trans results in the ligation of two protein or peptide segments linked to appropriate intein fragments. We have characterized the trans splicing reaction mediated by a naturally expressed, approximately 100-residue N terminal fragment of the Mycobacterium tuberculosis intein and a synthetic peptide containing the 38 C-terminal intein residues, and found that the splicing reaction was very versatile and robust. The efficiency of splicing was nearly independent of temperature between 4 and 37 degrees C and pH between 6.0 and 7.5, with only a slight decline at pH values as high as 8.5. In addition, there was considerable flexibility in the choice of the C-terminal intein fragment, no significant difference in protein ligation efficiency being observed between reactions utilizing the N-terminal fragment and either the naturally expressed 107-residue C-terminal portion of the intein, much smaller synthetic peptides, or the 107-residue C-terminal intein fragment modified by fusion of a maltose binding protein domain to its N-terminus. The ability to use different types of the C-terminal intein fragments and a broad range of reaction conditions make protein splicing in trans a versatile tool for protein ligation. PMID- 10685047 TI - Synthesis of a three zinc finger protein, Zif268, by native chemical ligation. AB - Protein synthesis by native chemical ligation has been an effective approach for the synthesis of proteins of moderate size. The utility of this approach for protein synthesis is demonstrated by the synthesis of a transcription factor, Zif 268 that contains three zinc finger domains. This synthesis highlights the modular nature of the chemical ligation approach and the ability to synthesize, handle and fold multiple domain proteins. PMID- 10685048 TI - A pH-tunable peptide ligase. AB - A chemical ligation system is reported, in which a highly acidic coiled-coil peptide was used to template two basic peptide fragments and catalyze their condensation, in a pH-tunable fashion, to generate a coiled-coil product. This template showed a high catalytic efficiency (with single turnover) under neutral conditions. Under acidic conditions, however, its catalytic efficiency was reduced by approximately 4500-fold. PMID- 10685049 TI - Synthesis, microbicidal activity, and solution structure of the dodecapeptide from bovine neutrophils. AB - The dodecapepetide sequence R-L-C-R-I-V-V-I-R-V-C-R with a disulfide bridge between the cysteine residues found in bovine neutrophils was synthesized by solid-phase procedures. Its antimicrobial activity against oral microorganisms such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus gordonii was examined, and its structural features were examined by CD and determined by two-dimensional (2D) nmr. The strains P. gingivalis (W50 and 381), A. actinomycetemcomitans (Y4 and 67), S. gordonii (DL1), and S. mutans (GS5) are found to be highly sensitive to this peptide at 2-2.5 microM concentrations, suggesting that the dodecapeptide is a potent antibiotic for oral pathogens. The weak negative n-sigma* band observed at approximately 265-270 nm in the CD spectra of this peptide provides evidence for the presence of a disulfide bridge. The negative n-pi* band at approximately 200 nm and the positive pi-pi* band at 185 nm suggest a folded structure for this peptide. The negative n-pi* shifts from 200 to 206 nm with an increase in intensity in dipalmitoylphosphotidylcholine vesicles, suggesting that the peptide might associate to form higher order aggregates in lipid medium. The assignment of backbone and side-chain proton resonances has been accomplished by the combined analysis of 2D total correlated and nuclear Overhauser effect spectroscopy. The temperature dependence of amide NH chemical shifts and (1)H (2)H exchange effect on amide NH resonances indicate the involvement of amide NH groups of Cys3, Ile5, Ile8, Val10, and Arg12 in intramolecular hydrogen bonding. The coupling constant (J(NH-C(alpha)H)) values, the set of medium-, short-, and long-range nuclear Overhauser effects, and the results of restrained structure calculation using the distance geometry algorithm for nmr applications provide evidence for a folded, loop-like structure with a type I (III) beta-turn involving Ile5, Val6, Val7, and Ile8, and two antiparallel beta-strands involving the N-terminal Arg1, Leu2, Cys3, and Val4 and the C-terminal Arg9, Val10, Cys11, and Arg12 residues. The structure of the dodecapeptide mimics the amphiphilic structure of large 30-35 residue defensins and the peptide appears to exhibit similar antimicrobial potency. PMID- 10685050 TI - Osmolyte-induced changes in protein conformational equilibria. AB - Examining solute-induced changes in protein conformational equilibria is a long standing method for probing the role of water in maintaining protein stability. Interpreting the molecular details governing the solute-induced effects, however, remains controversial. We present experimental and theoretical data for osmolyte induced changes in the stabilities of the A and N states of yeast iso-1 ferricytochrome c. Using polyol osmolytes of increasing size, we observe that osmolytes alone induce A-state formation from acid-denatured cytochrome c and N state formation from the thermally denatured protein. The stabilities of the A and N states increase linearly with osmolyte concentration. Interestingly, osmolytes stabilize the A state to a greater degree than the N state. To interpret the data, we divide the free energy for the reaction into contributions from nonspecific steric repulsions (excluded volume effects) and from binding interactions. We use scaled particle theory (SPT) to estimate the free energy contributions from steric repulsions, and we estimate the contributions from water-protein and osmolyte-protein binding interactions by comparing the SPT calculations to experimental data. We conclude that excluded volume effects are the primary stabilizing force, with changes in water-protein and solute-protein binding interactions making favorable contributions to stability of the A state and unfavorable contributions to the stability of the N state. The validity of our interpretation is strengthened by analysis of data on osmolyte-induced protein stabilization from the literature, and by comparison with other analyses of solute-induced changes in conformational equilibria. PMID- 10685051 TI - Similarities in the HIV-1 and ASV integrase active sites upon metal cofactor binding. AB - The HIV-1 integrase, which is essential for viral replication, catalyzes the insertion of viral DNA into the host chromosome thereby recruiting host cell machinery into making viral proteins. It represents the third main HIV enzyme target for inhibitor design, the first two being the reverse transcriptase and the protease. We report here a fully hydrated 2 ns molecular dynamics simulation performed using parallel NWChem3.2.1 with the AMBER95 force field. The HIV-1 integrase catalytic domain previously determined by crystallography (1B9D) and modeling including two Mg(2+) ions placed into the active site based on an alignment against an ASV integrase structure containing two divalent metals (1VSH), was used as the starting structure. The simulation reveals a high degree of flexibility in the region of residues 140-149 even in the presence of a second divalent metal ion and a dramatic conformational change of the side chain of E152 when the second metal ion is present. This study shows similarities in the behavior of the catalytic residues in the HIV-1 and ASV integrases upon metal binding. The present simulation also provides support to the hypothesis that the second metal ion is likely to be carried into the HIV-1 integrase active site by the substrate, a strand of DNA. PMID- 10685052 TI - Physicochemical characterization of generation 5 polyamidoamine dendrimers. AB - The dispersity, size, and self-interaction of generation 5 polyamidoamine dendrimeric polymers with different terminal groups (surfaces) were characterized using several physicochemical techniques. Amino-surface dendrimers form oligomeric aggregates in aqueous solution, even in the presence of high salt concentrations (0.6M sodium phosphate). In contrast, the hydroxyl-surface polymer G5-OH behaves as a single homogeneous (or paucidisperse) species at low concentration. Measurements of density increment and the sedimentation and diffusion coefficients of G5-OH suggest a more swollen, porous structure than a globular protein of comparable mass. Measurements of the concentration dependence of sedimentation equilibrium of G5-OH in pH 7.2 phosphate buffer indicate the presence of significant electrostatic repulsion overlaid on weakly attractive interactions, leading to the formation of nonspecific aggregates at sufficiently high dendrimer concentration. PMID- 10685053 TI - Condensation of DNA by multivalent cations: experimental studies of condensation kinetics. AB - DNA in viruses and cells exists in highly condensed, tightly packaged states. We have undertaken an in vitro study of the kinetics of DNA condensation by the trivalent cation hexaammine cobalt (III) with the aim of formulating a quantitative, mechanistic model of the condensation process. Experimental approaches included total intensity and dynamic light scattering, electron microscopy, and differential sedimentation. We determined the average degree of condensation, the distribution of condensate sizes, and the fraction of uncondensed DNA as a function of reaction time for a range of [DNA] and [Co(NH(3))(3+)(6)]. We find the following: (1) DNA condensation occurs only above a critical [Co(NH(3))(3+)(6)] for a given DNA and salt concentration. At the onset of condensation, [Co(NH(3))(3+)(6)]/[DNA-phosphate] is close to the average value of 0.54, which reflects the 89-90% charge neutralization criterion for condensation. (2) The equilibrium weight average hydrodynamic radius of the condensates first decreases, then increases with increasing [Co(NH(3))(3+)(6)] as they undergo a transition from intramolecular (monomolecular) to intermolecular (multimolecular) condensation. However, is insensitive to [DNA]. (3) The uncondensed DNA fraction decays approximately exponentially with time. The equilibrium uncondensed DNA fraction and relaxation time decrease with increasing [Co(NH(3))(3+)(6)] but are insensitive to [DNA]. (4) The condensation rate in its early stages is insensitive to [DNA] but proportional to [Co(NH(3))(3+)(6)](xs) = [Co(NH(3))(3+)(6)] - [Co(NH(3))(3+)(6)](crit). (5) Data for low [DNA] and low [Co(NH(3))(3+)(6)] at early stages of condensation are most reliable for kinetic modeling since under these conditions there is minimal clumping and network formation among separate condensates. A mechanism with initial monomolecular nucleation and subsequent bimolecular association and unimolecular dissociation steps with rate constants that depend on the number of DNA molecules in the condensate, accounts reasonably well for these observations. PMID- 10685054 TI - Low frequency vibrations and structural characterization of a murine IgG2a monoclonal antibody studied by raman and IR spectroscopies. AB - The Raman and ir spectra of a murine IgG2a monoclonal antibody molecule are reported. In accordance with previous studies on immunoglobulins, the secondary structure is predominantly of the beta-sheet type. The low frequency region of the Raman spectrum was also analyzed in detail. A structured band with two maxima near 43 and 94 cm(-1) was observed. This band has been attributed to vibrations of elastic body. The results are interpreted using a simple elastic model. PMID- 10685055 TI - Sequence-specific liquid crystallinity of collagen model peptides. I. Transmission electron microscopy studies of interfacial collagen gels. AB - The conformation, crystal structure and self-assembly behavior of three peptides with collagen-like repetitive sequences [(1) peptide GAPGPP: (Glu)(5)(Gly-Ala-Pro Gly-Pro-Pro)(6)(Glu)(5); (2) peptide GVPGPP: (Glu)(5)(Gly-Val-Pro-Gly-Pro Pro)(6)(Glu)(5); and (3) peptide GAPGPA: (Glu)(5)(Gly-Ala-Pro-Gly-Pro Ala)(6)(Glu)(5)] were compared. The peptides were characterized using transmission electron microscopy, electron diffraction, environmental scanning electron microscopy, and Fourier transform ir spectroscopy in order to determine how the molecular geometry dictated by each sequence affects the spontaneous generation of long-range ordered structures. Samples of each peptide, at ambient temperature and at 5 degrees C, were examined as films dried from aqueous solution, air-water interfacial films, and chloroform-water interfacial films. Peptide GAPGPP prepared at 5 degrees C and dried from bulk solution was found to have a collagen-like triple-helical structure. A sinusoidally textured gel, suggestive of cholesteric behavior was observed for peptides GAPGPP and GVPGPP at the aqueous chloroform interface at 5 degrees C. Peptide GAPGPA also formed a gel, but less reproducibly and the sinusoidal texture was not as well defined. The periodicities of the sinusoidal textures were reproducibly 10 microm for peptide GAPGPP, 7 microm for peptide GVPGPP, and 6 microm for peptide GAPGPA. The differences in the periodicity of the banded structure and in the crystallization behavior of the three peptides is attributed to differences in the symmetry of the preferred packing arrangement for each peptide, as evidenced by electron diffraction from crystallites that coexist with the sinusoidal gel. These differences are believed to be a measure of the effective symmetry and shape of the molecular cross section. PMID- 10685056 TI - Foreword PMID- 10685057 TI - Vascular pathology: a pathogenetic challenge for a new millennium. PMID- 10685058 TI - Atherosclerosis, inflammation, and infection. AB - In recent years, it has been shown that inflammation plays an important role in the pathogenesis of atherosclerosis. Activated macrophages, T lymphocytes, and mast cells are present in atherosclerotic plaques, which has led to the notion that the inflammatory response is an immune-mediated process. Complicated lesions, moreover, appear to be associated with an increase in the amount of the inflammatory response and in these patients, increased levels of acute phase proteins are present. The appreciation that atherosclerosis is an immune-mediated inflammatory disease has also led to renewed interest in the potential role of infectious agents in initiating or modulating atherosclerosis. Seroepidemiological studies have shown raised antibody titres against several micro-organisms. However, as yet, there are hardly any data available that provide a sound scientific basis for an infectious origin. Of all potential candidate organisms, Chlamydia pneumoniae appears as the one most likely involved in atherogenesis. C. pneumoniae has been retrieved from atherosclerotic tissues; the level of raised plasma titres correlates with the severity of symptomatic atherosclerotic disease; and the incidence of C. pneumoniae-responsive T cells in peripheral blood is increased in patients with coronary heart disease. It also appears that in some patients T cells generated from atherosclerotic plaques respond to C. pneumoniae. At the present state of knowledge, however, it is fair to state that the relationship between infection, intraplaque inflammation, and atherosclerosis still remains hypothetical, despite the increasing evidence that such a relationship could exist. PMID- 10685059 TI - Nitric oxide in the pathogenesis of vascular disease. AB - Nitric oxide (NO) is synthesized by at least three distinct isoforms of NO synthase (NOS). Their substrate and cofactor requirements are very similar. All three isoforms have some implications, physiological or pathophysiological, in the cardiovascular system. The endothelial NOS III is physiologically important for vascular homeostasis, keeping the vasculature dilated, protecting the intima from platelet aggregates and leukocyte adhesion, and preventing smooth muscle proliferation. Central and peripheral neuronal NOS I may also contribute to blood pressure regulation. Vascular disease associated with hypercholesterolaemia, diabetes, and hypertension is characterized by endothelial dysfunction and reduced endothelium-mediated vasodilation. Oxidative stress and the inactivation of NO by superoxide anions play an important role in these disease states. Supplementation of the NOS substrate L-arginine can improve endothelial dysfunction in animals and man. Also, the addition of the NOS cofactor (6R) 5,6,7, 8-tetrahydrobiopterin improves endothelium-mediated vasodilation in certain disease states. In cerebrovascular stroke, neuronal NOS I and cytokine inducible NOS II play a key role in neurodegeneration, whereas endothelial NOS III is important for maintaining cerebral blood flow and preventing neuronal injury. In sepsis, NOS II is induced in the vascular wall by bacterial endotoxin and/or cytokines. NOS II produces large amounts of NO, which is an important mediator of endotoxin-induced arteriolar vasodilatation, hypotension, and shock. PMID- 10685060 TI - Pathophysiology of ischaemia-reperfusion injury. AB - Reperfusion of ischaemic tissues is often associated with microvascular dysfunction that is manifested as impaired endothelium-dependent dilation in arterioles, enhanced fluid filtration and leukocyte plugging in capillaries, and the trafficking of leukocytes and plasma protein extravasation in postcapillary venules. Activated endothelial cells in all segments of the microcirculation produce more oxygen radicals, but less nitric oxide, in the initial period following reperfusion. The resulting imbalance between superoxide and nitric oxide in endothelial cells leads to the production and release of inflammatory mediators (e.g. platelet-activating factor, tumour necrosis factor) and enhances the biosynthesis of adhesion molecules that mediate leukocyte-endothelial cell adhesion. Some of the known risk factors for cardiovascular disease (hypercholesterolaemia, hypertension, and diabetes) appear to exaggerate many of the microvascular alterations elicited by ischaemia and reperfusion (I/R). The inflammatory mediators released as a consequence of reperfusion also appear to activate endothelial cells in remote organs that are not exposed to the initial ischaemic insult. This distant response to I/R can result in leukocyte-dependent microvascular injury that is characteristic of the multiple organ dysfunction syndrome. Adaptational responses to I/R injury have been demonstrated that allow for protection of briefly ischaemic tissues against the harmful effects of subsequent, prolonged ischaemia, a phenomenon called ischaemic preconditioning. There are two temporally and mechanistically distinct types of protection afforded by this adaptational response, i.e. acute and delayed preconditioning. The factors (e.g. protein kinase C activation) that initiate the acute and delayed preconditioning responses appear to be similar; however the protective effects of acute preconditioning are protein synthesis-independent, while the effects of delayed preconditioning require protein synthesis. The published literature in this field of investigation suggests that there are several potential targets for therapeutic intervention against I/R-induced microvascular injury. PMID- 10685061 TI - The role of apoptosis in vascular disease. AB - Normal arteries are characterized by a low turnover of endothelial (EC) and smooth muscle cells (SMC). Different mechanisms protect the EC and SMC against apoptosis in the normal artery. In hypertension, SMC replication is increased but this is not counterbalanced by increased apoptosis, resulting in thickening of the media of arteries and arterioles. The significance of apoptosis in atherosclerosis depends on the stage of the plaque, localization and the cell types involved. Both macrophages and SMC undergo apoptosis in atherosclerotic plaques. Apoptosis of macrophages is mainly present in regions showing signs of DNA synthesis/repair. SMC apoptosis is mainly present in less cellular regions and is not associated with DNA synthesis/repair. Even in the early stages of atherosclerosis SMC become susceptible to apoptosis since they increase different pro-apoptotic factors. Moreover, recent data indicate that SMC may be killed by activated macrophages. The loss of the SMC can be detrimental for plaque stability since most of the interstitial collagen fibres, which are important for the tensile strength of the fibrous cap, are produced by SMC. Apoptosis of macrophages could be beneficial for plaque stability if apoptotic bodies were removed. Apoptotic cells that are not scavenged in the plaque activate thrombin, which could further induce intraplaque thrombosis. It can be concluded that apoptosis in primary atherosclerosis is detrimental since it could lead to plaque rupture and thrombosis. Recent data of our group indicate that apoptosis decreased after lipid lowering which could be important in the understanding of the cell biology of plaque stabilization. PMID- 10685062 TI - Molecular mechanisms that control endothelial cell contacts. AB - Endothelial cell contacts control the permeability of the blood vessel wall. This allows the endothelium to form a barrier for solutes, macromolecules, and leukocytes between the vessel lumen and the interstitial space. Loss of this barrier function in pathophysiological situations can lead to extracellular oedema. The ability of leukocytes to enter tissue at sites of inflammation is dependent on molecular mechanisms that allow leukocytes to adhere to the endothelium and to migrate through the endothelial cell layer and the underlying basal lamina. It is a commonly accepted working hypothesis that inter-endothelial cell contacts are actively opened and closed during this process. Angiogenesis is another important process that requires well-controlled regulation of inter endothelial cell contacts. The formation of new blood vessels by sprouting from pre-existing vessels depends on the loosening of established endothelial cell contacts and the migration of endothelial cells that form the outgrowing sprouts. This review focuses on the molecular composition of endothelial cell surface proteins and proteins of the cytoskeletal undercoat of the plasma membrane at sites of inter-endothelial cell contacts and discusses the current knowledge about the potential role of such molecules in the regulation of endothelial cell contacts. PMID- 10685063 TI - Improving vascular grafts: the importance of mechanical and haemodynamic properties. AB - In the last 40 years, as techniques and materials have improved, the success rate of vascular prostheses with a diameter greater than 6mm has risen steadily, 5 year survival rates exceeding 95% in most centres. With smaller grafts no comparable improvement has occurred, the majority failing within 5 years, usually as a result of intimal hyperplasia and, ultimately atherosclerosis, in and around the downstream anastomosis. Clinical evidence suggests that the patency rates of small grafts are improved by matching the elastic properties of the graft to that of the artery into which it is placed. Although there is little reliable evidence that 'elastic mismatch' per se is the cause of intimal hyperplasia, it is generally accepted that mechanical factors are important in its genesis. These include disturbed flow at the anastomosis leading to fluctuations in shear stress at the endothelium (a known cause of intimal hyperplasia in normal arteries), injury due to suturing and stress concentration at the anastomosis. Few suitable materials or techniques have yet been developed to improve the long-term survival rates of small grafts. Recent advances in tissue engineering in which prostheses are manufactured by culturing vascular smooth muscle cells on a tubular scaffold of biodegradable polymer may ultimately make it possible to manufacture biologically and haemodynamically compatible grafts with diameters as small as 1mm. PMID- 10685064 TI - Molecular mechanisms in intimal hyperplasia. AB - Intimal hyperplasia is the process by which the cell population increases within the innermost layer of the arterial wall, such as occurs physiologically during closure of the ductus arteriosus and during involution of the uterus. It also occurs pathologically in pulmonary hypertension, atherosclerosis, after angioplasty, in transplanted organs, and in vein grafts. The underlying causes of intimal hyperplasia are migration and proliferation of vascular smooth muscle cells provoked by injury, inflammation, and stretch. This review discusses, at a molecular level, both the final common pathways leading to smooth muscle migration and proliferation and their (patho)-physiological triggers. It emphasizes the key roles played by growth factors and extracellular matrix degrading metalloproteinases, which act in concert to remodel the extracellular matrix and permit cell migration and proliferation. PMID- 10685065 TI - Interactions between cancer cells and the endothelium in metastasis. AB - The haematogenous phase of cancer metastasis facilitates the transport of metastatic cells within the blood and incorporates a sequence of interactions between circulating intravascular cancer cells and the endothelium of blood vessels at the sites of tumour cell arrest. Initial interactions involve mechanical contact and transient adhesion, mediated by endothelial selectins and their ligands on the neoplastic cells. This contact initiates a sequence of activation pathways that involves cytokines, growth factors, bioactive lipids, and reactive oxygen species produced by either the cancer cell or the endothelium. These molecules elicit expression of integrin adhesion molecules in cancer cells and the endothelium, matrix metalloproteinases, and chemotactic factors that promote the attachment of tumour cells to the vessel wall and/or transvascular penetration. Induction of endothelial free radicals can be cytotoxic to cancer cells. Collectively, the sum of these interactions constitutes an interdependent relationship, the outcome of which determines the fate of the metastatic process. PMID- 10685066 TI - The clinical manipulation of angiogenesis: pathology, side-effects, surprises, and opportunities with novel human therapies. AB - The first phase of angiogenesis research has provided knowledge of the basic pathobiology of angiogenesis and its manipulation in models, mouse, and man. The first line of therapeutic substances has been devised and is now in clinical trials. New lessons are being learned from clinical observations. Unexpected side effects are being noted, particularly affecting the nervous system. Other side effects may be anticipated from a sound knowledge of clinical pathology and recognition of the commonality of angiogenesis to multiple disease mechanisms, but these may be tolerable or avoidable. Angiogenesis researchers await further feedback and ideas from the clinic to stimulate the next phase of basic and applied research. PMID- 10685067 TI - The pathophysiology of the collateral circulation (arteriogenesis). AB - Since the mid 1980s a new strategy is coming from bench to bedside termed angiogenesis. This process involves sprouting of capillaries and finally results in newly developed microvessels which belong to the capillary level. Importantly these newly formed capillary tubes lack vascular smooth muscle cells, they are not surrounded by mural cells and are fragile and prone to rupture. Therefore these networks remain susceptible to hypoxic regulation, fail to become remodelled and are unable to sustain proper circulation: they cannot adapt to changes in physiological demands of blood supply. Since atherosclerosis affects large conductance arteries, capillary sprouting from compromised vessels cannot provide an adequate supply of blood flow to the endangered tissue. However, the body provides a natural system of pre-existing collateral arteries, which may bypass sites of arterial occlusion. These vessels can dramatically increase their lumen by growth so as to provide enhanced perfusion to the jeopardized ischaemic regions. This process - termed arteriogenesis - finally results in fully functional and structurally normal arteries which can ameliorate the ensuing detrimental effects of vessel obstruction in many regions of the body. Hallmarks of arteriogenesis are increased levels of shear forces (rather than ischaemia), the invasion of circulating monocytes (and their pluripotent precursors), and the substrates of arteriogenesis are pre-existing collateral arterioles. PMID- 10685068 TI - Regulation of inflammatory vascular damage. AB - The acute inflammatory response is comprised of an elaborate cascade of mediators that control an ordered sequence of events resulting in the recruitment of neutrophils to the site of infection or injury. Microvascular injury occurring during acute inflammation often results in increased vascular permeability and microvascular haemorrhage. Damage to vascular endothelial cells, basement membrane, and matrix components results from both neutrophil-dependent and neutrophil-independent mechanisms and is also dependent on the organ/tissue source of the endothelial cells. Neutrophil-mediated injury of endothelial cells involves a complex cascade in which products from both cell types affect the cytotoxic outcome. It is also clear that the acute inflammatory response is carefully regulated by the endogenous gene expression of both pro-inflammatory and anti-inflammatory mediators. Control of acute inflammation seems to relate to activation of the transcription factor NFkappaB. To appreciate the interrelationship between multiple contributing factors of inflammatory vascular injury, one must first have an understanding of the inflammatory mediator cascades which bring about the recruitment of neutrophils to the site of inflammation. In this review it is discussed how inflammatory mediators, as well as the products of activated neutrophils, affect the outcome of the acute inflammatory response. PMID- 10685069 TI - Pathogenesis of ANCA-associated systemic vasculitis. AB - The aetiology of primary systemic vasculitides remains unknown. Recent advances have been made in the understanding of relevant mechanisms of inflammation, particularly the role of the endothelium and interactions with inflammatory mediators and immune effector cells. In Wegener's granulomatosis and microscopic polyangiitis the evidence suggests an autoimmune inflammatory process, characterized by an early lesion involving neutrophils and endothelial cells as both targets and active participants; priming of neutrophils and endothelial cells allows ANCA to activate neutrophils with damage localized to the endothelium. In the absence of immune complex deposition, the role of the ANCA is particularly intriguing. Endothelial cell damage and activation produces pro inflammatory mediators with influx monocytes and T cells intensifying damage. Increased understanding of the pathogenesis of systemic vasculitis is likely to provide the basis for the use of more selective immunomodulatory therapies in the future. PMID- 10685070 TI - Vascular protease receptors: integrating haemostasis and endothelial cell functions. AB - The endothelium plays a crucial dynamic role as a protective interface between blood and the underlying tissues during the haemostatic process, which maintains blood flow in the circulation and prevents life-threatening blood loss. Following vessel wall injury with initial platelet adhesion and aggregation to exposed subendothelial extracellular matrix, the initiation, amplification, and control of haemostasis depend on structurally unrelated membrane-associated receptors for blood coagulation proteases including tissue factor, G-protein-coupled protease activatable receptors, thrombomodulin, and protein C receptor, respectively. In addition to their regulatory role in haemostasis, the respective (pro-)enzyme ligands such as Factors VIIa and Xa, thrombin or protein C mediate specific signalling pathways in vascular cells related to migration, proliferation or adhesion. The functional importance of these receptors beyond haemostasis has been manifested by various lethal and pathological phenotypes in knock-out mice. These protease receptors thereby provide important molecular links in the vascular system and serve to integrate haemostasis with endothelial cell functions which are relevant for the (patho-)physiological responses to injury or inflammatory challenges. PMID- 10685071 TI - Microcirculatory dysfunction in sepsis: a pathogenetic basis for therapy? AB - Sepsis is a frequent complication of multiple organ dysfunction syndrome and remains a major problem of intensive care medicine. It is also a common factor in the final cause of death in hospital populations. Clinical observations, assisted by invasive monitoring techniques as well as pathological-anatomical studies, clearly indicate that microcirculatory dysfunction lies at the centre of sepsis pathogenesis. Numerous animal models, from rodents to primates, many of which employ bacteria or their toxins, especially endotoxins, have helped to shed light on the pathomechanisms leading to this dysregulation in the peripheral circulation. Among these are activation of humoral and cellular inflammatory mediator systems, with special emphasis on neutrophil-endothelial interactions, affecting endothelial barrier function and vasoregulation and ultimately leading to severely perturbed oxygen transport and utilization. In vitro studies have provided more insight into the molecular mechanisms involved in this microcirculatory dysfunction, although much more attention must be directed towards microvascular endothelial cells and the role of heterogeneity of response in various vascular beds. These experimental data must in turn be validated by comparing with the human in situ situation, both clinical and morphological. This review aims at a critical appraisal of the clinical and experimental evidence for sepsis-induced dysregulation of the microcirculation and how knowledge of the underlying cellular and molecular pathology could be used to make therapy more rational and effective. To date, therapeutic approaches, such as anti-cytokine and anti-oxidant regimens, which have been highly successful in experimental models, have failed to demonstrate clinical efficacy. Newer approaches, such as targeting the coagulation system, nitric oxide synthesis or intracellular signal transduction, are also discussed. The necessity to focus on the role of anti inflammatory mediators, as well as the pathogenetic significance of important molecular groups, such as the heat shock proteins, which until now have been given scant attention, will be stressed. PMID- 10685072 TI - Transgenic mouse models in angiogenesis and cardiovascular disease. AB - Novel gene technologies have allowed us to manipulate the genetic balance of candidate molecules in mice in a controllable manner. Homologous or site-specific recombination in embryonic stem cells allows us to study the consequences of deficiencies, mutations, and conditional or tissue-specific expression of gene products in transgenic mice. These technological breakthroughs have significantly advanced biomedical research and broadened our understanding of the pathophysiological role of candidate disease genes. In addition, gene transfer allows us to test the possible therapeutic use of gene products for gene therapy. A variety of assays have been miniaturized, allowing analysis of cardiovascular physiology in the mouse. With the advent of genome sequencing programmes, these gene technologies provide means of studying gene function in a conclusive manner. Furthermore, disease models can be generated which can be used as test models for (gene) therapy or for the discovery of novel genes using differential gene profiling techniques. The present review will focus on the molecular basis of how blood vessels form (angiogenesis and arteriogenesis) and how they become diseased. A selected number of molecules that have been studied in the authors' laboratory will be reviewed in more detail. PMID- 10685073 TI - Cytometric quantification of nitrate reductase by immunolabeling in the marine diatom Skeletonema costatum. AB - BACKGROUND: The uptake of nitrate by phytoplankton is a central issue in biological oceanography due to its importance to primary production and vertical flux of biogenic carbon. Nitrate reductase catalyzes the first step of nitrate assimilation, the reduction of NO(3) to NO(2). A cytometric protocol to detect and quantify relative changes in nitrate reductase (NR) protein content of the marine centric diatom Skeletonema costatum is presented. METHODS: Immunolabeling of NR protein was achieved with polyclonal antibodies raised against S.costatum NR. Antisera specific to a NR protein subunit and to a NR polypeptide sequence were compared, and cytometric results of NR protein abundance were related to Western analyses. Changes in cellular NR abundance and activity were followed during an upwelling simulation experiment in which S. costatum was exposed to a shift from ammonia to nitrate as major nitrogen source. RESULTS: NR protein could be detected in NO(3)-grown cells and at extremely low levels hardly discernible by Western Blot densiometry in NH(4)-grown cells. The protocol allowed observation of early stages of NR induction during an upwelling simulation. NR abundance increased after the nutrient shift to reach a new physiological "steady state" 96 hrs later. NR activity exhibited diel variation with maxima at mid-day. NR abundance as estimated by both flow cytometry and Western analysis exhibited a hyperbolic relationship to NR activity. This pattern suggests post-translational activation of NR protein. CONCLUSIONS: The presented protocol allows the differentiation of NH(4)- versus NO(3)-grown algae as well as the monitoring of early stages in the induction of nitrate assimilatory capacities. PMID- 10685074 TI - On the use of the Kolmogorov-Smirnov statistical test for immunofluorescence histogram comparison. AB - BACKGROUND: The problem considered is the quantitative comparison of immunofluorescence frequency distributions in order to detect their differences of biological significance, i.e., to evaluate the potential positivity of a cell sample with respect to negative control cells. The Kolmogorov-Smirnov (KS) statistical test, proposed in the literature for this purpose, is examined and discussed through its application to a set of experimental measurements. It is shown that even differences due to the stain procedure or to instrumental biases may be considered significant by the test implemented in the standard form. METHODS: In order to ensure valid results, it is necessary to take into account the various sources of variation in the specific experimental context. A procedure is proposed that uses the KS statistics as a reference for determining an appropriate estimate of the overall variability in the control data. This estimate is derived from the comparisons of the cumulative distributions associated with repeated measurements of the negative cell sample. RESULTS AND CONCLUSIONS: The KS-related index thus defined provides a tool for assessing the potential positivity of a cell sample, since it allows to distinguish between statistical and biological significance of the difference between the histogram to be tested and the set of control data. In particular, if a cell sample is not included in the control variability, either a positive cell subpopulation is present, or all cells are positive. Instead, for a sample included in the control variability, the difference will be not biologically meaningful, even if statistically significant. Moreover, when a purely positive control sample is also available, it is possible to derive a measure of the precision at which a true biological positivity can be detected. Finally, since the index is not absolute, but relative to the features of the instrumentation, of the antibodies and of the fluorochromes used, it represents a quantitative measure of the stability and reproducibility of the measurement process and could be used for quality control of flow cytometric experiments in immunofluorescence. PMID- 10685075 TI - Differential production of IFN-gamma, analyzed at the single-cell level, by specific subsets of human NK and T cells from healthy and HIV(+) subjects. AB - BACKGROUND: Interferon gamma is a cytokine that plays a central role in immunity, and is physiologically secreted by T and NK cells under appropriate stimuli during the immune response. By means of flow cytometry, we performed a single cell analysis of interferon gamma producing NK cells and their surface phenotype in normal and HIV(+) individuals that show several defects of cytokine production and cellular immunity. METHODS: PBMC or purified NK cells were stimulated for 1 12 h with PMA/ionomycin in the presence of monensin, subsequently stained for surface CD56 and CD3 or CD8, and for intracytoplasmic IFN-gamma, and analysed by flow cytometry. RESULTS: Our results show that CD56(+) NK cells are more efficient interferon gamma producers than T cells. Moreover, within the CD56(+) NK cell population, those that co-express low density CD8 are the best producers. Finally, we show that NK cells during HIV infection are more massively recruited to interferon gamma production than those from normal subjects. CONCLUSIONS: Both in the normal and HIV(+) subjects, a higher percentage of NK cells than T cells can produce IFN-gamma although differences can be identified within the NK cells subset in terms of IFN-gamma production. The production of IFN-gamma is fully achievable in the HIV(+) subjects, which is consistent with their elevated plasmatic levels of the cytokine. The possibility that NK cells that produce interferon gamma could represent a functionally distinct population committed to the production of this cytokine, is discussed. PMID- 10685076 TI - Flow cytometric evaluation of fas expression in relation to response and resistance to anthracyclines in leukemic cells. AB - BACKGROUND: Cell chemosensitivity to cytotoxic drugs has been attributed to their ability to trigger apoptosis. The emergence of resistance in drug-exposed cells is often characterized by the appearance of drug efflux mechanisms including P-gp transport. Nevertheless, mdr1 expression may coexist with other resistance features, in particular those interfering with apoptotic signaling pathways. METHODS: Leukemic cell lines cultured in a progressively toxic environment were analyzed for Fas and P-gp expression by immunostaining and flow cytometry. Their mdr1 mRNA expression level was determined by reverse transcriptase-polymerase chain reaction (RT-PCR), and their apoptotic response was microscopically evaluated. Activation of the Fas pathway was obtained by cross-linking the Fas receptor with the 7C11 anti-Fas agonist. RESULTS: We demonstrate a dose-dependent Fas overexpression after short-term (18 h) incubation with daunorubicin. The subsequent sensitization to Fas activators led to a significant increase in the apoptotic response induced by 7C11. After long-term exposure to daunorubicin and acquisition of drug resistance, expression of P-gp was accompanied by a decrease in the number of Fas sites at the cell surface with a correlated desensitization to Fas-induced apoptosis. Additional alterations in the Fas signaling pathway can also be hypothesized in the most resistant Jurkat cell line. CONCLUSIONS: The induction of Fas expression could be one of the mechanisms of action of chemotoxic drugs and thus might enhance the cell susceptibility to Fas-mediated apoptosis. On the contrary, the emergence of the multidrug resistance phenotype is associated with a down-regulation of Fas expression and possible defects in the Fas signaling pathway. PMID- 10685078 TI - KRN5500, a novel antitumor agent, induces apoptosis or cell differentiation in HL 60 cells. AB - BACKGROUND: KRN5500, a derivative of spicamycin, shows antitumor activity against a variety of tumor cell lines. However, the mechanism of cytotoxic action has remained unclear. METHODS: The viability of HL-60 human leukemic cells treated with KRN5500 was studied by the dye exclusion assay. Induction of apoptosis and effects on the cell cycle were investigated by flow cytometry: We measured cellular DNA content after extraction of fragmented DNA, and apoptosis-induced DNA strand breaks. Cell morphology was observed by light microscopy. DNA strand breaks at a nucleosomal unit were analyzed by electrophoresis. RESULTS: Our data demonstrated that KRN5500 caused inhibition of cell growth, and that apoptosis was the mode of cell death. G(1) phase cells were more susceptible to KRN5500 induced apoptosis. In addition, KRN5500 induced cell differentiation at lower concentration. CONCLUSIONS: It is anticipated that KRN5500 will be used clinically as an anti-leukemic agent. Its mechanism of antitumor action is to induce apoptosis or cell differentiation. PMID- 10685077 TI - Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green, and MitoTracker Green is affected by mitochondrial membrane potential altering drugs. AB - BACKGROUND: We set out to develop an assay for the simultaneous analysis of mitochondrial membrane potential and mass using the probes 10-nonyl acridine orange (NAO), MitoFluor Green (MFG), and MitoTracker Green (MTG) in HL60 cells. However, in experiments in which NAO and MFG were combined with orange emitting mitochondrial membrane potential (DeltaPsi(m)) probes, we found clear responses to DeltaPsi(m) altering drugs for both probes. METHODS: The three probes were titrated to determine whether saturation played a role in the response to drugs. The effects of a variety of DeltaPsi(m) altering drugs were tested for MFG and MTG at probe concentrations of 20 nM and 200 nM and for NAO at 0.1 microM and 5 microM, using rhodamine 123 at 0.1 microM as a reference probe. RESULTS: Incubation of GM130, HL60, and U937 cells with 2,3-butanedione monoxime (BDM), nigericin, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), carbonyl cyanide p (trifluoromethoxy)phenylhydrazone (FCCP), 2,4-dinitrophenol (DNP), gramicidin, ouabain, and valinomycin resulted in increases of the fluorescence intensity for MFG or MTG with only a few exceptions. The fluorescence intensity of cells stained with 0.1 microM NAO increased following incubation with BDM, nigericin, and decreased for FCCP, CCCP, DNP, gramicidin, and valinomycin. The results with 5 microM NAO were similar. CONCLUSIONS: MFG, MTG, and NAO appeared poor choices for the membrane potential independent analysis of mitochondrial membrane mass. Considering the molecular structure of these probes that favor accumulation in the mitochondrial membrane because of a positive charge, our results are not surprising. Cytometry 39:203-210, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10685079 TI - A novel deep red/low infrared fluorescent flow cytometric probe, DRAQ5NO, for the discrimination of intact nucleated cells in apoptotic cell populations. AB - BACKGROUND: The linking of intracellular metabolism of anticancer drugs with cellular response is problematic. We describe a new probe for cellular integrity, based upon a structure which has the additional potential to act as a substrate for cytochrome P450-dependent bioreductive metabolism. DRAQ5NO is an N-oxide modified anthraquinone with optimal fluorescence excitation maxima compatible with He-Ne (633 nm) and Kr-Ar (647 nm) lasers. METHODS: DRAQ5NO-loading and Annexin V binding was monitored using dual-laser flow cytometry (488 nm/633 nm wavelengths) in human lymphoma cultures undergoing anticancer drug- (etoposide; VP-16) induced apoptosis. RESULTS: DRAQ5NO gave an Em(lambdamax) of 700.5 nm but retains DNA binding potential with an emission wavelength red-shift of approximately 12 nm. The agent showed reduced cytotoxicity and a limited capacity to accumulate within cells compared with the non-N-oxide form that shows a high nuclear targeting capacity in intact cells. DRAQ5NO/Annexin V provides for a positive discrimination between intact cells, membrane-compromised cells, cellular debris, and early stage apoptotic cells. CONCLUSIONS: The spectral properties of DRAQ5NO allow for the use of visible range fluorochromes and differential excitation in multilaser systems for tracking apoptotic populations with implications for the measurement of bioreductive potential in complex tumour populations simultaneously undergoing physiologically or drug-induced apoptosis. PMID- 10685081 TI - Purification of human tonsil plasma cells: pre-enrichment step by immunomagnetic selection of CD31(+) cells. AB - BACKGROUND: The advancement of knowledge about the biology of human normal plasma cells (PC) is hampered by their low frequency and difficult isolation. The aim of this study is to design a way of purifying these cells. METHODS: To this end, advantage was taken of the fact that human tonsil PC expressed surface CD31 at higher levels than the rest of tonsil B cells. RESULTS: The immunomagnetic selection of CD31(+) cells from tonsil B cells increased by a factor of 12 the proportion of PC, determined as CD38(high) cells. This method recovered half of the initial number of PC, and did not alter the PC functions, because IgG secretion was similar in control B cell cultures as well as in cultures of B cells obtained at successive steps of the selection procedure. In addition, CD38(high) cells pre-enriched by this technique were readily isolated by FACS sorting and clearly identified as PC. CONCLUSIONS: Therefore, immunomagnetic pre enrichment of CD31(+) cells is an efficient method that allows the complete purification of human functional PC. PMID- 10685080 TI - Fetal nucleated erythrocyte recovery: fluorescence activated cell sorting-based positive selection using anti-gamma globin versus magnetic activated cell sorting using anti-CD45 depletion and anti-gamma globin positive selection. AB - BACKGROUND: Fluorescence activated cell sorting (FACS)-based anti-gamma (gamma) positive selection and magnetic activated cell sorting (MACS)-based anti-CD45 depletion followed by anti-gamma positive staining have been two of the most frequently used methods to isolate fetal cells from maternal blood. To date, there has been no direct comparison of fetal cell recovery by these two methods. This study was designed to address this issue. METHODS: Fluorescence in situ hybridization (FISH) was performed on nucleated anti-gamma positive cells using X and Y probes. Twenty-four maternal blood samples were obtained immediately after elective termination of pregnancy to ensure a detectable number of fetal cells. RESULTS: The yield and purity of fetal nucleated erythrocytes (FNRBCs) was statistically higher in FACS sorted samples (P < 0.01). The specificity of staining for FNRBCs was statistically higher in MACS sorted samples (P < 0.01). CONCLUSIONS: The data from this study demonstrate that both techniques have benefits and limitations. FACS has the advantage of having higher yield, higher purity, higher FISH efficiency and ease in microscope analysis, and MACS has the advantage of having higher specificity and less cell loss during FISH. PMID- 10685082 TI - Accurate quantitation of Leishmania infection in cultured cells by flow cytometry. AB - BACKGROUND: Leishmaniases are major parasitic diseases caused by protozoans that are obligate intracellular parasites during the mammalian phase of their life cycle. Quantitation of experimental mammalian cell infections is usually performed by time-consuming microscopic examination. In this report a flow cytometry (FCM)-based assay suitable for studying in vitro infections by L.amazonensis is presented. METHODS: Intense fluorescence staining of the amastigote forms with a stage- and species-specific monoclonal antibody was obtained after permeabilization of both the host-cell cytoplasmic membrane and the parasitophorous vacuole membrane by saponin treatment. RESULTS: Upon flow cytometry (FCM) analysis, parasitized cells separated sharply from the auto fluorescence of the mammalian host cells, giving the assay a high degree of sensitivity and specificity. Ninety to 98% of cells in the more fluorescent population harbored parasites visible by phase-contrast and UV-light microscopy, while no parasites were observed in more than 95% of the cells in the population with background fluorescence. Comparisons of the FCM results with those from microscope counting and analysis of various dilutions of parasitized cells confirmed the reliability of the method. CONCLUSIONS: The FCM assay provided rapid quantitation of Leishmania infection either in mouse macrophages, the natural host cell in murine leishmaniasis, or in Chinese hamster ovary (CHO) cells, a non-macrophage cell line proposed as an in vitro model for studying host parasite interactions. The protocol described here should be adaptable to studies involving other parasites residing in nucleated cells. PMID- 10685083 TI - In this issue. Dermatologic laser surgery. PMID- 10685084 TI - Recent developments in cutaneous lasers. PMID- 10685085 TI - Imaging superficial tissues with polarized light. AB - OBJECTIVE: Polarized light can be used to obtain images of superficial tissue layers such as skin, and some example images are presented. This study presents a study of the transition of linearly polarized light into randomly polarized light during light propagation through tissues. STUDY DESIGN/MATERIALS AND METHODS: The transition of polarization was studied in polystyrene microsphere solutions, in chicken muscle (breast) and liver, and in porcine muscle and skin. The transition is discussed in terms of a diffusion process characterized by an angular diffusivity (radians(2)/mean free path) for the change in angular orientation of linearly polarized light per unit optical path traveled by the light. RESULTS: Microsphere diffusivity increased from 0.031 to 0.800 for diameters decreasing from 6.04 microm to 0.306 microm, respectively. Tissue diffusivity varied from a very low value (0.0004) for chicken liver to an intermediate value (0.055) for chicken and porcine muscle to a very high value (0.78) for pig skin. CONCLUSION: The results are consistent with the hypothesis that birefringent tissues randomize linearly polarized light more rapidly than nonbirefringent tissues. The results suggest that polarized light imaging of skin yields images based only on photons backscattered from the superficial epidermal and initial papillary dermis because the birefringent dermal collagen rapidly randomizes polarized light. This anatomic region of the skin is where cancer commonly arises. PMID- 10685086 TI - Evaluation of cooling methods for laser dermatology. AB - BACKGROUND AND OBJECTIVE: Skin cooling is used to protect the epidermis in a variety of laser dermatology procedures, including leg vein treatment, hair removal, and port wine stain removal. Spray and contact cooling are the two most popular methods, but similarities and differences of these techniques are not well understood. STUDY DESIGN/MATERIALS AND METHODS: A theoretical model of skin cooling is presented for two different regimens: "soft" cooling in which freezing of the skin is not permitted and "hard" cooling in which the skin can be frozen to a given depth. Spray and contact cooling were also compared experimentally using an in vitro model. RESULTS: For a fixed skin surface temperature, spray and contact cooling theoretically produce the same cooling profile in the skin. Anatomic depth of cooling depends on the time for which either the spray or contact is applied. In vitro experiments caused temperature at the simulated basal layer to be between -5 and +5 degrees C for both spray (tetrafluoroethane, boiling point -26 degrees C) and contact (-27 degrees C sapphire plate) cooling. The theoretical precooling analysis shows hard mode to be faster and more selective than soft mode; however, cooling time for hard mode must be carefully controlled to prevent irreversible epidermal damage caused by freezing. CONCLUSIONS: Both spray and contact cooling provide efficient skin cooling. The choice of cooling method depends on other factors such as the target depth, cost, safety, and ergonomic factors. PMID- 10685087 TI - Bioheat transfer analysis of cryogen spray cooling during laser treatment of port wine stains. AB - BACKGROUND AND OBJECTIVE: The thermal response of port wine stain (PWS) skin to a combined treatment of pulsed laser irradiation and cryogen spray cooling (CSC) was analyzed through a series of simulations performed with a novel optical thermal model that incorporates realistic tissue morphology. STUDY DESIGN/MATERIALS AND METHODS: The model consisted of (1) a three-dimensional reconstruction of a PWS biopsy, (2) a Monte Carlo optical model, (3) a finite difference heat transfer model, and (4) an Arrhenius thermal damage calculation. Simulations were performed for laser pulses of 0.5, 2, and 10 ms and a wavelength of 585 nm. Simulated cryogen precooling spurts had durations of 0, 20, or 60 ms and terminated at laser onset. Continuous spray cooling, which commenced 60 ms before laser onset and continued through the heating and relaxation phases, was also investigated. RESULTS: The predicted response to CSC included maximal pre irradiation temperature reductions of 27 degrees C at the superficial surface and 12 degrees C at the dermoepidermal junction. For shorter laser pulses (0.5, 2 ms), precooling significantly reduced temperatures in superficial regions, yet did not effect superficial vessel coagulation. Continuous cooling was required to reduce significantly thermal effects for the 10-ms laser pulse. CONCLUSIONS: For the PWS morphology and treatment parameters studied, optimal damage distributions were obtained for a 2-ms laser pulse with a 60-ms precooling spurt. Epidermal and vascular morphology as well as laser pulse duration should be taken into account when planning CSC/laser treatment of PWS. Our novel, realistic-morphology modeling technique has significant potential as a tool for optimizing PWS treatment parameters. PMID- 10685088 TI - New approaches to the treatment of vascular lesions. AB - BACKGROUND AND OBJECTIVE: The pulsed dye laser was developed based on the concept of selective photothermolysis. By using a wavelength of light well absorbed by the target and pulse duration short enough to spatially confine thermal injury, specific vascular injury could be produced. STUDY DESIGN/MATERIALS AND METHODS: Although the pulsed dye laser revolutionized the treatment of port wine stains (PWS) and a variety of other vascular lesions, the ideal thermal relaxation time for the vessels in PWS is actually 1-10 ms, not 450 micros of the original pulsed dye laser machines. These original theoretical calculations recently have been proven correct in a study that used both an animal vessel model and in human PWS. RESULTS: Longer wavelengths of light, within the visible spectrum, penetrate more deeply into the skin and are more suitable for deeper vessels, whereas longer pulse durations are required for larger caliber vessels. CONCLUSION: A variety of lasers recently have been developed for the treatment of vascular lesions which incorporate these concepts into their design, including pulsed dye lasers at 1.5 ms, a filtered flash-lamp pulsed light source with pulse durations of 1-20 ms, several 532-nm pulsed lasers with pulse durations of 1 ms to as high as 100 ms, long pulsed alexandrite lasers at 755 nm with pulse durations up to 20 ms, pulsed diode lasers in the 800 to 900 nm range, and long pulsed 1064 Nd:YAG sources. PMID- 10685089 TI - Mathematical modeling for the prediction and optimization of laser hair removal. AB - BACKGROUND AND OBJECTIVE: The study of hair removal is a slow, tedious process. Efficacy evaluations require test-site observation for at least one complete hair cycle, a minimum of 6-8 months. In addition, tracking and counting individual hairs is extremely labor intensive. The objective of this study was to develop and evaluate a mathematical model for hair removal that could significantly speed the entire process. STUDY DESIGN/MATERIALS AND METHODS: Generally accepted kinetic and statistical modeling methods were used to develop a mathematical description of hair growth. The anagen and telogen percentages and decay times were the variables used to predict the kinetics of untreated hair. In the case that the follicles were treated, it was necessary to additionally consider the possible outcomes after treatment, making the calculations much too complicated for simple mathematical formulations. Therefore, a computerized statistical model was developed that considered the probabilities of no, partial, or complete follicular damage in addition to the untreated model variables. These models were then evaluated by comparing them to data derived from the literature and a study center. RESULTS: Values derived from the mathematical model were capable of closely approximating the experimental results of untreated (shaving) and treated (plucking, electrolysis, ruby laser, Q-switched Nd:YAG laser) hair growth kinetics. The model was also shown to be useful for optimizing the number and interval of Q-switched Nd:YAG laser treatments. CONCLUSIONS: A mathematical model can be used to reliably predict results from a variety of hair removal techniques. It also appears to be useful for optimizing a particular treatment protocol. In addition, the development of new hair removal products may be aided by using this method. PMID- 10685090 TI - Ruby laser hair removal: evaluation of long-term efficacy and side effects. AB - BACKGROUND AND OBJECTIVE: Although several studies on laser-assisted hair removal have been published, data on long-term follow-up are few. The present study investigated the long-term efficacy and safety of normal-mode ruby laser pulses on hair removal. STUDY DESIGN/MATERIALS AND METHODS: The normal-mode ruby laser (Epilaser; 694 nm, 3 msec) was used to treat a wide range of body sites in 51 volunteers. The mean follow-up after the last treatment was 8.37 months. RESULTS: Sixty-three percent of the patients had sparse regrowth. The mean fluence used was 46.5 J/cm(2) in patients who had sparse hair regrowth and 39.3 J/cm(2) in patients who had moderate hair regrowth (P = 0.0127). Transient pigmentary changes occurred most frequently in patients with skin type 4. CONCLUSION: The normal-mode ruby laser is an efficient and safe method for long-term hair reduction, especially in fair-skinned individuals with dark hair. Higher fluences produce greater long-term efficacy. Adverse effects are minimal and transient. PMID- 10685091 TI - Nonablative skin remodeling: selective dermal heating with a mid-infrared laser and contact cooling combination. AB - BACKGROUND AND OBJECTIVE: Many of the microscopic changes associated with photodamage reside in the dermis. It follows that subsurface heating of the skin might allow for cosmetic enhancement without loss of the epidermis. Accordingly, we investigated the clinical and microscopic changes produced by a mid-infrared laser coupled with a contact cooling device. STUDY DESIGN/MATERIALS AND METHODS: Nine patients were treated with an erbium glass laser and sapphire cooling handpiece in contact with the skin. Postauricular sites were irradiated with pulse energies varying from 400-1,200 mJ and numbers of pulses from 4 to 40. Outcome measures included pain, edema, and erythema at predetermined postoperative intervals. Biopsies were performed just after treatment and 2 months postoperatively for selected pulse energy-pulse number combinations. RESULTS: Erythema, edema, and pain increased with pulse energy and number of pulses. Likewise, immediate epidermal necrosis and subsequent scarring were observed for larger pulse energy-pulse number combinations. At sites with epidermal preservation, on biopsy, immediate dermal thermal damage was observed in a band-like pattern. The deep boundaries of this band were dependent on pulse energy and pulse number. After 8 weeks, biopsies showed dermal fibroplasia roughly correlating to the band of immediate dermal thermal damage. CONCLUSION: Selective dermal heating can be achieved with a mid-infrared laser coupled to a contact surface cooling device. In this study, the range of fibroplasia and lack of clinically substantial cosmetic enhancement suggest that the dermal thermal damage achieved may be too deep and that the injury should be confined to more superficial levels to alter the most severely photodamaged dermis. Lasers Surg. Med. 26:186-195, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10685092 TI - Nonablative treatment of rhytids with intense pulsed light. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the efficacy and complication rate of a nonablative nonlaser light source in the treatment of rhytids. Laser resurfacing, in the treatment of facial rhytids, has involved ablative methods, with their associated complications and limitations. Rhytid improvement requires dermal collagen remodeling. Interest has begun to focus on the use of wavelengths that preserve the epidermis but deliver enough energy to promote rhytid improvement. STUDY DESIGN/MATERIALS AND METHODS: Thirty subjects with class I-II rhytids and Fitzpatrick skin types I-II were treated with up to four treatments with an intense pulsed light source. Subjects were evaluated 6 months after the final treatment. RESULTS: Twenty-five subjects showed some improvement in the quality of skin. No subjects were found to have total resolution of rhytids. CONCLUSION: Nonlaser intense pulsed light may effectively improve some facial rhytids. Such improvement can occur without epidermal ablation. PMID- 10685093 TI - Prophylactic fluconazole promotes reepithelialization in full-face carbon dioxide laser skin resurfacing. AB - BACKGROUND AND OBJECTIVE: Laser skin resurfacing is used to treat photodamaged skin, rhytids, and acne scarring. Many patients are placed on antibiotics and antivirals pre- and postoperatively. The purpose of this study was to determine whether prophylactic fluconazole increased the rate of reepithelialization in patients undergoing full-face CO(2) laser skin resurfacing. STUDY DESIGN/MATERIALS AND METHODS: Ninety-one patients underwent full-face CO(2) laser skin resurfacing with the Coherent Ultrapulse 5000C. At least two passes of 300 mJ, density 5, were used except periocularly. Study group I consisted of 48 consecutive patients who received either cephalexin or ciprofloxacin for 7 days postoperatively. Study group II consisted of 43 patients who received 300 mg of fluconazole on postoperative days 3-8, in addition to ciprofloxacin. Both groups received acyclovir 400 mg t.i.d. pre- and postoperatively. RESULTS: Time to complete reepithelialization was compared between the groups by t-test. Group II reepithelialized significantly faster than group I (7.65 +/- 1.20 days vs. 10.27 +/- 2.94 days; P < 0.0001). Ninety-five percent of patients receiving fluconazole (group II) healed completely by day 9 versus only 53% of patients in group I. CONCLUSION: Fluconazole administered postoperatively between days 3 and 8 significantly promotes reepithelialization in patients undergoing full-face CO(2) laser skin resurfacing. PMID- 10685094 TI - Variable pulse erbium:YAG laser skin resurfacing of perioral rhytides and side-by side comparison with carbon dioxide laser. AB - BACKGROUND AND OBJECTIVE: Laser resurfacing of facial rhytides has become a popular treatment option for many patients with wrinkles, photoaging, and acne scarring. Laser wavelength/pulse duration options and new techniques continue to shorten the healing phase associated with laser skin resurfacing while maintaining clinical efficacy. Variable pulse erbium:YAG (Er:YAG) laser systems are now available that offer the surgeon the ability to vary the Er:YAG pulse duration from a pulse that is primarily ablative to one that is more thermal. The objective of this study was to evaluate the histologic effects created with a variable pulse Er:YAG laser. To study prospectively the clinical effects on upper lip rhytides with a variable pulse Er:YAG laser when compared side by side with pulsed carbon dioxide (CO(2)) laser resurfacing. STUDY DESIGN/MATERIALS AND METHODS: Forty-two treatment sites on 21 patients were randomized and evaluated after treatment of the upper lip region with CO(2) laser resurfacing on one side and a variable pulse Er:YAG laser on the other. Patient diaries were maintained to assess erythema, crusting, pain, and pigmentary changes. Blinded objective grading of improvement was performed. Chromometer measurements were obtained to analyze erythema. RESULTS: The variable pulse Er:YAG laser treatment reduced the duration of crusting on average from 7.7 days with CO(2) to 3.4 days. Chromometer measurements noted decreased postoperative erythema. Grading by physicians in a blinded manner showed 63% improvement for the CO(2) treatment site and 48% improvement in the variable pulse Er:YAG site. No cases of permanent hyperpigmentation, hypopigmentation, or scarring occurred. CONCLUSION: The variable pulse Er:YAG laser resurfacing is a safe and effective resurfacing tool, which combines ablative and thermal modalities. The protocol used in this study approaches but does not equal the results we have traditionally seen with CO(2) laser resurfacing. PMID- 10685095 TI - Deep coagulation of dermal collagen with repetitive Er:YAG laser irradiation. AB - BACKGROUND AND OBJECTIVE: Er:YAG lasers are known to effectively ablate human skin with minimal thermal damage to subjacent dermal tissue. We have investigated whether deep coagulation of dermal collagen, similar to that observed with the CO(2) laser, could be achieved with repetitive Er:YAG laser exposures. STUDY DESIGN/MATERIALS AND METHODS: Skin on the back of a Sprague-Dawley rat in vivo was irradiated with sequences of 1-10 Er:YAG laser pulses at a repetition rate of 10 or 33 Hz and single-pulse fluences from 0.8 to 1.4 J/cm(2). The resulting lesions were biopsied within 1 hour after laser exposure, and the histologic sections were examined by using optical microscopy. RESULTS: The depth of dermal collagen denaturation increases dramatically when 3-10 low-fluence Er:YAG laser pulses are stacked at a repetition rate of 10 or 33 Hz. CONCLUSION: Coagulation of dermal collagen deeper than 200 microm below the epidermal-dermal junction is feasible by using the appropriate settings of a repetitive Er:YAG laser. PMID- 10685096 TI - Modulating the Er:YAG laser. AB - BACKGROUND AND OBJECTIVES: In the past 2 years, there has been some controversy about the optimal laser system, or combination of systems, for cutaneous resurfacing. Initially, it seemed that the Er:YAG laser would have significant advantages over the CO(2) laser. In practice, some of those who jumped early onto the Er:YAG bandwagon have been unimpressed with the degree of skin tightening that can be achieved with this system. Also, the excessive bleeding induced by the Er:YAG lasers prevented deeper vaporization. During the past 18 months, three new "modulated" Er:YAG lasers have been produced that are said to be able to achieve CO(2) laser-like effects, while maintaining the Er:YAG laser advantages. The purpose of this article is to examine these new systems and to discuss their potential benefits, if any, over the "conventional" Er:YAG lasers, and the CO(2) lasers. STUDY DESIGN/MATERIALS AND METHODS: The author has collected data from his own experience and that of his colleagues in the department of dermatology at University of California at San Francisco. The author has used all three types of modulated Er:YAG laser on patients presenting for cosmetic laser resurfacing and the treatment of many benign conditions over an 18-month period. RESULTS: All three modulated forms of Er:YAG lasers have been demonstrated to provide better coagulation than the conventional Er:YAG lasers. The Derma-K and the Contour Er:YAG lasers were able to induce tissue contraction/desiccation similar to the CO(2) laser. The author and his colleagues have induced only two cases of permanent hypopigmentation in over 50 cases during the past 18 months while using the Er:YAG laser, significantly less than might be expected with the CO(2) lasers. CONCLUSIONS: If a laser surgeon is happy with the results obtained with a high-energy, short-pulse CO(2) laser, then there seems little reason to consider changing to an Er:YAG laser. The modulated Er:YAG lasers have definite advantages over the conventional Er:YAG lasers. They exhibit better control of hemostasis and can ablate tissue to a greater depth than the conventional Er:YAG lasers. The Er:YAG lasers might induce less permanent hypopigmentation than the CO(2) lasers. PMID- 10685097 TI - Fatty acid incorporation by Rhodnius prolixus midgut. AB - [(14)C]Oleic acid injected into the hemocoel of Rhodnius prolixus females was shown to rapidly associate with lipophorin particles. Half of the lipophorin associated [(14)C]oleic acid was transferred in about 5 min to different organs, but the midgut was the main organ to take it up on day 10 after a blood meal. The rate of [(14)C]oleic acid incorporation by the midgut was high up to 15 min after injection and then declined. The [(14)C]oleic acid incorporated by the midgut was found in phospholipids (58.6%) and neutral lipids (37.4%). The midgut capacity to incorporate [(14)C]oleic acid varied on different days after a meal: it increased up to day 10 and then decreased. The fate of the [(14)C]lipids synthesized by the midgut was followed and it was observed that 10 days after feeding diacylglycerol was the main lipid released to hemolymph and that most of phospholipids and triacylglycerols remained associated with the midgut. The metabolism of free fatty acids in Rhodnius prolixus females is discussed in the context of major biological events that follow a blood meal such as digestion and oogenesis. PMID- 10685098 TI - Identification of cuticular hydrocarbons and the alkene precursor to the pheromone in hemolymph of the female gypsy moth, Lymantria dispar. AB - Hydrocarbons were extracted from the surface of the cuticle and from the hemolymph of adult female gypsy moths. GC and GC/MS analysis indicated that the cuticular hydrocarbons with chain lengths >21 carbons were the same as those found in the hemolymph. These consisted of mostly saturated straight chain hydrocarbons with heptacosane the major component. Methyl branched hydrocarbons were also identified including a series of tetramethylalkanes with chain lengths of 30, 32, and 34 carbons. In addition to those found on the cuticle surface, the hemolymph contained the alkene pheromone precursor, 2-methyl-Z7-octadecene and two saturated analogues, 2-methyl-octadecane and 2-methyl-hexadecane. No evidence was obtained for the presence of the pheromone 2-methyl-7, 8-epoxy-octadecane in the hemolymph. Pheromone gland extracts indicated that small amounts (<1 ng) of the alkene precursor were also present in the gland. Relatively larger amounts of the alkene precursor were found in the hemolymph at the time when pheromone titers were higher on the gland. The presence of the hydrocarbon pheromone precursor in the hemolymph is discussed in relation to possible biosynthetic pathways for producing the gypsy moth pheromone. PMID- 10685099 TI - Characterization of proteases from a stored product mite, Tyrophagus putrescentiae. AB - Extracts of Tyrophagus putrescentiae feces exhibited higher (>50-fold) specific protease activity rates than those measured using mite body extracts for the substrates azocasein, BApNa, SA(2)PPpNa, HA, and HPA. This suggests that trypsin, chymotrypsin, and carboxypeptidases A and B are involved in mite digestion. Hydrolysis of the substrates ZAA(2)MNA and LpNa was only 3 times higher in fecal extracts, suggesting that levels of cathepsin B and aminopeptidases in the lumen of the digestive tract are low compared to the other enzymes. The hydrolysis of hemoglobin was only detected in body extracts indicating that cathepsin D is not a digestive protease in this species. Protease inhibitors of different specificity were tested invivo to establish their potential as control agents. We found that development from larvae to adult was significantly retarded in larvae fed on brewers' yeast containing inhibitors of serine proteases, whereas no such effect was found with inhibitors of cysteine and aspartyl proteases. Interestingly, when dietary mixtures of serine protease, aminopeptidase and carboxypeptidase inhibitors were fed to T.putrescentiae, a synergistic effect was observed that retarded development. Several plant lectins were also tested, but none affected development. PMID- 10685100 TI - Periodicity of sex pheromone biosynthesis, release and degradation in the lightbrown apple moth, Epiphyas postvittana (Walker). AB - Pheromone titer in moths is a product of three processes occurring in or at the surface of the pheromone gland: biosynthesis, release, and intraglandular degradation, of pheromone. Changes in titers of sex pheromone, the fatty acyl pheromone analog (FAPA), and tetradecanoate, a pheromone biosynthetic intermediate, were studied in detail in the lightbrown apple moth, Epiphyas postvittana (Walker). Although changes in the pheromone titers in a day were relatively small, with the peak titer being 2-3 times greater than that at the trough, pheromone titer did show a distinct diel periodicity. Titer of the FAPA showed a similar, but less variable, diel pattern, but tetradecanoate titer showed little or no diel pattern. The pattern of pheromone titer suggested that females biosynthesize pheromone at two different rates during the photoperiod: a high rate during the latter half of the photophase and most of the scotophase, which is associated with a high pheromone titer, and a low rate throughout the first half of the photophase, which is associated with a low titer. Consistent with data on commencement of copulation, pheromone was released from the second hour of the scotophase through to the eighth hour. Pheromone release rate during this period appeared to be similar to the rate of pheromone biosynthesis. In contrast to the other two processes, pheromone degradation did not appear to have a diel pattern. Females decapitated at different times of the photoperiod showed a similar decline in pheromone titer, consistent with the reaction kinetics being first order in pheromone titer. PMID- 10685101 TI - Artificial reproduction in a hymenopteran insect, Athalia rosae, using eggs matured with heterospecific yolk proteins and fertilized with cryopreserved sperm. AB - Previtellogenic ovaries of Athalia rosae (Hymenoptera, Symphyta, Tenthredinidae) were transplanted into the adult female abdominal hemocoel of Athalia infumata (Symphyta, Tenthredinidae), Arge nigrinodosa (Symphyta, Argidae), and Pimpla nipponica (Apocrita, Ichneumonidae). The donor oocytes accumulated heterospecific yolk proteins and matured in the A. infumata host. On average, six mature oocytes were obtained per transplanted ovary. In contrast, the donor oocytes accumulated a limited amount of yolk but did not mature in the Ar. nigrinodosa host and did not even accumulate yolk in the P. nipponica host. The eggs that matured in the A. infumata host were injected with cryopreserved A. rosae sperm that had been taken from adult male seminal vesicles and stored at -80 degrees C. Fertilization, as confirmed by the use of visible marker mutations, was achieved and a fraction of the injected eggs developed into fertile female adults. PMID- 10685102 TI - Pesticide residues in medicinal plants and phytomedicines. AB - Pesticides, which are mainly applied on crops for the protection of plants against a range of pests, have been found in crude medicinal plants as well as in infusions, decoctions, tinctures and essential oils. This fact has caused concern in various segments of society and scientific investigation has been demanded to assess the health hazards more accurately. The present review covers more than 30 years (1963-1998) of published methods of analysing pesticide residues in medicinal plants, with special emphasis on the relevance of these matrices, the legislation, the risks involved in using material containing uncontrolled amounts of residues and the possible effects of technological factors on the proportion of pesticide transferred from the raw material to the end product. PMID- 10685103 TI - Antiviral effect of flavonoids on the dengue virus. AB - In the present study we analysed the possible antiviral effect on dengue viruses of different flavonoids extracted and identified at the Chemistry Institute, UNAM, from the Mexican plants Tephrosia madrensis, Tephrosia viridiflora and Tephrosia crassifolia. The flavonoids glabranine and 7-O-methyl-glabranine presented 70% inhibition on the dengue virus at a concentration of 25 microM, while methyl-hildgardtol A, hildgardtol A and elongatine had no effect on viral growth. Our results show that glabranine and 7-O-methyl-glabranine isolated from Tephrosia s.p. exert a dose-dependent inhibitory effect in vitro on the dengue virus. PMID- 10685104 TI - Phenylpropanoids from Ballota nigra L. inhibit in vitro LDL peroxidation. AB - From the European plant Ballota nigra L. various polyphenols including phenylpropanoid derivatives were isolated. There is increasing evidence that oxidized low-density lipoproteins (Ox-LDL) might be involved in the pathogenesis of atherosclerosis and it has been reported that polyphenols inhibit LDL peroxidation and atherogenesis. The goal of this study was to test whether the major polyphenolic compounds extracted from Ballota nigra, four phenylpropanoid glycosides, verbascoside, forsythoside B, arenarioside, and ballotetroside and one non-glycosidic phenylpropanoid, caffeoyl-L-malic acid, inhibit Cu(2+)-induced LDL peroxidation. The effectiveness of these compounds was compared to the activity of quercetin, a well-known polyphenol inhibitor of Cu(2+)-induced LDL oxidation. Antioxidant efficacious doses (ED 50) of arenarioside and ballotetroside were 1.8 microM and 7.5 microM respectively, while in the same conditions, the ED 50 of forsythoside B and verbascoside were similar (1 microM) and those of quercetin and of caffeoyl-L-malic acid were 2.3 microM and 9.5 microM respectively. Spectrophotometric studies show that quercetin is a Cu(2+) chelator while phenylpropanoid glycosides and caffeoyl-L-malic acid are not Cu(2+) chelators. Therefore, phenylpropanoid glycosides are strong inhibitors of Cu(2+)-induced LDL oxidation, independent of any capacity to act as Cu(2+) chelators. PMID- 10685105 TI - Toxicological evaluation of the hydro-alcohol extract of the dry leaves of Peumus boldus and boldine in rats. AB - The hydro-alcohol extract of the dry leaves of Peumus boldus and boldine, showed abortive and teratogenic action and changes in the blood levels of bilirubin, cholesterol, glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and urea in rats. The long term administration of the extract and boldine did not cause histological modification during a period of 90 days. PMID- 10685106 TI - Hypoglycaemic action of the flavonoid fraction of Cuminum nigrum seeds. AB - The seeds of Cuminum nigrum were screened phytochemically and were found to contain 8% flavonoids and 0.01% alkaloids. When studied for their effect on blood glucose levels, oral administration of the flavonoid contents of the plant caused a hypoglycaemic effect at a dose range of 0.5 to 1.5 g/kg, both in normal and alloxan-diabetic rabbits. The hypoglycaemic effect started 2 h after drug administration, reaching a maximum within 4-8 h and the blood glucose levels returned close to normal within 24 h of drug administration. The glibenclamide (5 mg/kg), produced a hypoglycaemic effect in the normal rabbits, whereas it had no effect on the blood glucose levels of alloxan-diabetic rabbits. The alkaloids isolated from C. nigrum seeds, however, failed to exert any significant hypoglycaemic effect in either the normal or diabetic rabbits. A 7 day acute toxicity study in rabbits did not produce any apparent adverse effect at doses as high as 5 g/kg orally. These data indicate that the total flavonoid contents of C. nigrum seeds exhibited considerable hypoglycaemic activity in rabbits and may therefore be responsible for the previously reported antidiabetic activity of the seeds. Furthermore, it is conceivable that the C. nigrum flavonoids possess insulin triggering and/or insulin-like properties. PMID- 10685107 TI - Effect of Psidium guajava leaves on some aspects of the central nervous system in mice. AB - The present work examines the effects of hexane, ethyl acetate and methanol extracts of Psidium guajava leaves (20,100,500 and 1250 mg/kg) on the central nervous system in mice. The three extracts exhibited mostly dose-dependent antinociceptive effects in chemical and thermal tests of analgesia. The extracts also produced dose-dependent prolongation of pentobarbitone-induced sleeping time. However, they had variable and mostly non-significant effects on locomotor coordination, locomotor activity or exploration. In the pharmacological tests used, the ethyl acetate extract seemed to be the most active, followed by the hexane and then the methanol extracts. PMID- 10685108 TI - Antibacterial activity of diospyrin, isodiospyrin and bisisodiospyrin from the root of Diospyros piscatoria (Gurke) (Ebenaceae). AB - Two dimeric naphthoquinones, diospyrin and isodiospyrin, isolated from the root of Diospyros piscatoria (Gurke), a common ingredient in several folk medicines, have been shown to have a broad spectrum of antibacterial activity. The minimum inhibitory concentrations (MICs) of diospyrin against Streptococcus pyogenes ATCC 12344 and Streptococcus pneumoniae ATCC 33400 ranged from 1.56 to 50 microg/mL. While those against Salmonella choleraesuis serotype typhi (S. typhi), ATCC 6539 and Mycobacterium chelonae ATCC 19977 were between 25 and 100 microg/mL. Isodiospyrin was more active than its racemic isomer diospyrin. The MICs against Gram-positive bacteria ranged from 0.78 to 50 microg/mL. While those against Pseudomonas aeruginosa ATCC 15443 and S. typhi ranged from 50 to 100 microg/mL. The MIC for M. chelonae was between 6.25 and 25 microg/mL. MICs were found to increase with the concentration of cells used for the inoculum. The MICs for Bacillus subtilis ATCC 6633 increased up to the highest concentration of cells tested. The same phenomenon was observed on M. chelonae, but with better effect in the latter. The kinetics of bacteria studies against both B. subtilis and M. chelonae increases with increasing concentration of isodiospyrin tested. Two tetrameric forms of plumbagin were isolated. The naphthoquinone bisisodiospyrin, gave MIC values between 300 and 400 micro g/mL. The second, as yet unidentified tetramer, was not active at 500 micro g/mL. PMID- 10685109 TI - Evaluation of antibacterial activity of Asparagus racemosus willd. root. AB - Different concentrations (50, 100, 150 microg/mL) of the methanol extract of the roots of Asparagus racemosus Willd. showed considerable in vitro antibacterial efficacy against Escherichia coli, Shigella dysenteriae, Shigella sonnei, Shigella flexneri, Vibrio cholerae, Salmonella typhi, Salmonella typhimurium, Pseudomonas putida, Bacillus subtilis and Staphylococcus aureus. The effects produced by the methanol extract were compared with chloramphenicol. PMID- 10685111 TI - Adrenocorticosterone alterations in male, albino mice treated with Trichopus zeylanicus, Withania somnifera and Panax ginseng preparations. AB - The levels of corticosterone were estimated by the HPLC method in the adrenal glands of stressed (5 h constant swimming) male albino mice treated with Trichopus zeylanicus, Withania somnifera and Panax ginseng preparations and compared with non-treated stressed and normal controls. The treatments increased the corticosterone levels in all the groups. The physical endurance (increased survival time) of swimming mice also increased in all the treated groups, except in the group treated with Withania somnifera powder (500 mg/kg, p.o.). PMID- 10685110 TI - Effect of Ocimum sanctum roots extract on swimming performance in mice. AB - The effect of a methanol extract, obtained from the roots of Ocimum sanctum, on mouse swimming performance were studied using three different doses. On the basis of our findings, a high dose (400 mg/kg, i.p.) of the extracts of Ocimum sanctum increased the swimming time suggesting a central nervous system stimulant and/or antistress activity. The effect produced by the extract was comparable to that of desipramine, an antidepressant drug. PMID- 10685112 TI - Screening of Mediterranean Rosaceae plants for their molluscicidal and piscicidal activities. AB - Extracts of increasing polarity of 13 plants belonging to Mediterranean Rosaceae were tested for possible lethal toxicity against Biomphalaria glabrata Say, a snail intermediate host of Schistosoma mansoni Sambon. Due to the search for compounds without toxicity in nontarget organisms, principally fish and humans, the piscicidal toxicity of the active extracts was also evaluated. PMID- 10685113 TI - Effects of the water soluble fraction from leaves of Ageratum conyzoides on smooth muscle. AB - The water soluble fraction of Ageratum conyzoides L. (WSF) was studied in isolated rat uterus and intestinal smooth muscles in order to evaluate its popular use as a spasmolytic. WSF (0.2 and 0.4 mg/mL) increased EC(50) values and decreased maximum responses to acetylcholine and calcium chloride. WSF (0.5-3.3 mg/mL) also produced direct myorelaxant effect on smooth muscle preparations. Theophylline (10(-3) M) potentiated the relaxant action of WSF. Theophylline also prevented the decrease in maximum response promoted by WSF in acetylcholine concentration-effect curves. These results seem to be partially linked to calcium mobilization. The data also suggest that WSF could act synergistically with theophylline in the inhibition of cyclic AMP phosphodiesterase. The results give support to the popular medicinal indications of the plant. PMID- 10685114 TI - Toxicological studies on stem bark, leaf and seed kernel of yellow oleander (Thevetia peruviana). AB - A comparative study of the toxic effects of extracts from stem bark, leaf and seed kernel of yellow oleander (Thevetia peruviana) in albino rats was carried out. Male and female albino rats weighing 150-200 g were administered crude aqueous extracts of stem bark, leaf and seed kernel of the plant by intraperitoneal injection or exposed to baits prepared with the dry extracts of the plant parts. The control groups either received distilled water by injection, or were fed non-poisoned baits. Extracts from all the plant parts were toxic, and produced marked poisoning symptoms that culminated in death. Poisoning symptoms manifested earlier (10 min after treatment) in rats administered aqueous kernel extracts intraperitoneally as against 45 min to several hours in rats poisoned by ingestion of toxicant. Poisoning symptoms indicated serious cardiac, neuromotor and mental malfunctioning, and manifested as tachycardia, arrhythmia, paralysis, ataxia and disorientation. The lethal dose was lowest (507 mg/kg) with the concentrated aqueous kernel extract (CAKE), and highest (5700 mg/kg) with the bait formulated using 40% of the kernel meal - FKM(B). Rats treated by injection with aqueous kernel extract (AKE) died faster within 10 h, than those with the aqueous leaf or stem bark extracts that died after 260 h. No mortality or abnormal behavioural changes were observed among animals in the control groups. PMID- 10685115 TI - Antihyperglycaemic activity of Musa sapientum flowers: effect on lipid peroxidation in alloxan diabetic rats. AB - Musa sapientum commonly known as 'banana' is widely used in Indian folk medicine for the treatment of diabetes mellitus. Oral administration of 0.15, 0.20 and 0.25 g/kg body weight of the chloroform extract of the flowers for 30 days resulted in a significant reduction in blood glucose and glycosylated haemoglobin and an increase in total haemoglobin. The extract prevented a decrease in body weight, and also resulted in a decrease in free radical formation in the tissues. Thus the study shows that banana flower extract (BFEt) has an antihyperglycaemic action. The decrease in thiobarbituric acid reactive substances (TBARS) and the increase in reduced glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) clearly shows the antioxidant property of BFEt. The effect of BFEt was more prominently seen in the case of animals given 0.25 g/kg body weight. BFEt was more effective than glibenclamide. PMID- 10685117 TI - Patents alert PMID- 10685116 TI - The effect of reserpine, a modulator of multidrug efflux pumps, on the in vitro activity of tetracycline against clinical isolates of methicillin resistant Staphylococcus aureus (MRSA) possessing the tet(K) determinant. AB - As part of a screening programme to identify modulators of multidrug efflux in methicillin resistant Staphyloccocus aureus (MRSA), we have validated our assays using the antihypertensive plant alkaloid reserpine. Clinical isolates of MRSA were resistant to tetracycline and shown to possess the tet(K) determinant which encodes for the Tet(K) efflux protein, which conferred high level resistance to tetracycline (MIC = 128 microg/mL). In the presence of reserpine, a known inhibitor of multidrug resistance (mdr) efflux pumps, this MIC was significantly reduced (MIC = 32 microg/mL). PMID- 10685118 TI - Selected bibliography PMID- 10685119 TI - Evidence-based art of the clinical examination PMID- 10685120 TI - Effect of hormone replacement therapy on the bone mass and urinary excretion of pyridinium cross-links. AB - CONTEXT: The menopause accelerates bone loss and is associated with an increased bone turnover. Bone formation may be evaluated by several biochemical markers. However, the establishment of an accurate marker for bone resorption has been more difficult to achieve. OBJECTIVE: To study the effect of hormone replacement therapy (HRT) on bone mass and on the markers of bone resorption: urinary excretion of pyridinoline and deoxypyridinoline. DESIGN: Cohort correlational study. SETTING: Academic referral center. SAMPLE: 53 post-menopausal women, aged 48-58 years. MAIN MEASUREMENTS: Urinary pyr and d-pyr were measured in fasting urine samples by spectrofluorometry after high performance liquid chromatography and corrected for creatinine excretion measured before treatment and after 1, 2, 4 and 12 months. Bone mineral density (BMD) was measured by dual energy X-ray absorptiometry (DEXA) before treatment and after 12 months of HRT. RESULTS: The BMD after HRT was about 4.7% (P < 0.0004); 2% (P < 0.002); and 3% (P < 0. 01) higher than the basal values in lumbar spine, neck and trochanter respectively. There were no significant correlations between pyridinium cross-links and age, weight, menopause duration and BMD. The decrease in pyr and d-pyr was progressive after HRT, reaching 28.9% (P < 0.0002), and 42% (P < 0.0002) respectively after 1 year. CONCLUSIONS: Urinary pyridinoline and deoxypyridinoline excretion decreases early in hormone replacement therapy, reflecting a decrease in the bone resorption rate, and no correlation was observed with the bone mass evaluated by densitometry. PMID- 10685121 TI - Prognostic factors in non-Hodgkin lymphomas. AB - CONTEXT: In Hodgkin's disease, each clinical or pathologic stage can be related to the extent of the area involved and predicts the next anatomical region at risk for tumor dissemination. OBJECTIVE: To determine the best prognostic factors that could predict survival in non-Hodgkin lymphoma cases. DESIGN: A retrospective study. LOCATION: Department of Hematology and Transfusion Medicine, Universidade Federal de Sao Paulo - Escola Paulista de Medicina. PARTICIPANTS: 142 patients with non-Hodgkin lymphoma diagnosed between February 1988 and March 1993. MAIN MEASUREMENTS: Histological subset, Sex, Age, Race, B symptoms, Performance status, Stage, Extranodal disease, Bulk disease, Mediastinal disease, CNS involvement, BM infiltration, Level of DHL, Immunophenotype. RESULTS: In the first study (113 patients), the following variables had a worse influence on survival: yellow race (P<0.1); ECOG II, III e IV (P<0.1) and extranodal disease (P<0.1) for high grade lymphomas; constitutional symptoms (P<0.1), ECOG II, III e IV (P<0.1) and involvement of CNS (P<0.1) for intermediate grade and the subtype lymphoplasmocytoid (P=0.0186) for low grade lymphomas. In the second survey (93 patients), when treatment was included, the variables related to NHL survival were: CNS involvement (P<0.1) for high grade lymphomas, constitutional symptoms (P<0.1), ECOG II, III, IV (P=0.0185) and also CNS involvement (P<0.1) for the intermediate group. There were no variables related to the survival for low-grade lymphomas. CONCLUSIONS: The intermediate grade lymphomas were more compatible with data found in the literature, probably because of the larger number of patients. In this specific case, the treatment did not have an influence on the survival. PMID- 10685122 TI - Bilateral carotid body paraganglioma: case report. AB - CONTEXT: Surgical treatment of carotid body paragangliomas is a challenge to the surgeon because of the large vascularization of the tumor, involvement of the carotid vessels and the close anatomical relationship with the cranial nerves. CASE REPORT: A 63-year-old patient was submitted to resection of two carotid body paraganglioma tumors found in the right-side and left-side carotid bodies at the bifurcation of the common carotid arteries. Two surgeries were performed at different times and neither of them presented any morbidity. Arteriography was fundamental for diagnosis of the small, asymptomatic tumor on the right side. DESIGN: Case Report PMID- 10685123 TI - Cystic struma ovarii: a rare presentation of an infrequent tumor. AB - CONTEXT: Struma ovarii, a rare neoplasm, is a monophyletic teratoma composed of thyroid tissue. It is generally considered to account for less than 5% of mature teratomas. CASE REPORT: A diagnosis of struma ovarii may be the source of many diagnostic problems. It may be cystic and microscopic examination may only reveal a few typical thyroid follicles, resulting in confusion with other cystic ovarian tumors. Extensive sampling should be undertaken and immunohistochemistry may be decisive in establishing the thyroid nature of the epithelial lining. The authors report two cases of cystic struma ovarii, and discuss diagnostic criteria and the limitations of frozen biopsies in these tumors. PMID- 10685124 TI - The gut at war: the consequences of enteropathogenic Escherichia coli infection as a factor of diarrhea and malnutrition. AB - Diarrheal disease is still the most prevalent and important public health problem in developing countries, despite advances in knowledge, understanding, and management that have occurred over recent years. Diarrhea is the leading cause of death in children under 5 years of age. The impact of diarrheal diseases is more severe in the earliest periods of life, when taking into account both the numbers of episodes per year and hospital admission rates. This narrative review focuses on one of the major driving forces that attack the host, namely the enteropathogenic Escherichia coli (EPEC) and the consequences that generate malnutrition in an early phase of life. EPEC serotypes form dense microcolonies on the surface of tissue-culture cells in a pattern known as localized adherence (LA). When EPEC strains adhere to epithelial cells in vitro or in vivo they cause characteristic changes known as Attaching and Effacement (A/E) lesions. Surface abnormalities of the small intestinal mucosa shown by scanning electron microscopy in infants with persistent diarrhea, although non-specific, are intense enough to justify the severity of the clinical aspects displayed in a very young phase in life. Decrease in number and height of microvilli, blunting of borders of enterocytes, loss of the glycocalyx, shortening of villi and presence of a mucus pseudomembrane coating the mucosal surface were the abnormalities observed in the majority of patients. These ultrastructural derangements may be due to an association of the enteric enteropathogenic agent that triggers the diarrheic process and the onset of food intolerance responsible for perpetuation of diarrhea. An aggressive therapeutic approach based on appropriate nutritional support, especially the utilization of human milk and/or lactose-free protein hydrolyzate-based formulas and the adequate correction of the fecal losses, is required to allow complete recovery from the damage caused by this devastating enteropathogenic agent. PMID- 10685125 TI - Modification of high blood pressure after myocardial infarction. AB - The treatment of high blood pressure (BP) after myocardial infarction is extremely important to decrease reinfarction and mortality. BP should be controlled more strictly in this high-risk hypertensive population. Recently, many clinical trials have demonstrated the benefits of lifestyle modification and antihypertensive agents, particularly beta-blockers and angiotensin-converting enzyme inhibitors for the treatment of acute myocardial infarction. Treatment with these agents that modify BP may benefit even normotensive patients after a myocardial infarction, although the benefit is greater in hypertensives. PMID- 10685126 TI - Management of hypercholesterolemia. AB - Benefit from the treatment of hyperlipidemia has now been conclusively documented, and this article has focused on the clinical trial data supporting diet and drug therapy in adult patients with different lipoprotein disorders and discussed therapeutic approaches with a focus on reducing plasma concentrations of LDL cholesterol. National guidelines for the use of hypolipidemic drugs are strongly supported by the clinical trials and have appropriately set lower target concentrations of LDL cholesterol for patients with established atherosclerosis or diabetic patients as compared with patients with more than two cardiovascular risk factors or, the lowest risk group, patients without evidence of atherosclerosis and fewer than two known cardiovascular risk factors. The goals of therapy in patients with established atherosclerosis are to prevent further progression and potentially induce regression, whereas in high-risk patients (e.g., those with heterozygous familial hypercholesterolemia) without evidence of atherosclerosis, the aims of therapy are to reduce LDL cholesterol to a concentration at which subclinical atherosclerosis and xanthomas regress and the patient does not develop premature cardiovascular disease. Evidence-based medicine strongly supports clinical benefit from the treatment of hypercholesterolemia in men and women with and without known coronary artery disease, and the main goal should be ensure that patients who could benefit from lipid-lowering therapy are effectively treated and followed to ensure long-term compliance, efficacy, and safety. PMID- 10685127 TI - Management of selected lipid abnormalities. Hypertriglyceridemia, low HDL cholesterol, lipoprotein(a), in thyroid and renal diseases, and post transplantation. AB - Although the focus in treating lipid disorders is on reducing LDL cholesterol levels, triglycerides, HDL cholesterol, and Lp(a) are all independent risk factors that can be used clinically to assess cardiovascular risk. Decisions to initiate drug therapy for LDL cholesterol reduction may be influenced by levels of these other lipoprotein fractions. Data supporting intervention to modify these factors is less abundant than for LDL cholesterol reduction, but in certain circumstances drug therapy targeted at triglycerides or HDL cholesterol may be appropriate. Patients with nephrotic syndrome and end-stage renal disease are at particularly high risk for the development of cardiovascular disease and should be treated aggressively for lipid disorders. Finally, solid organ transplant recipients are almost always hyperlipidemic, and appropriate therapy could reduce cardiovascular events. PMID- 10685128 TI - Methods to enhance smoking cessation after myocardial infarction. AB - The evidence linking smoking and coronary artery disease is quite strong, and there is also a positive relationship between smoking cessation and reduction in cardiovascular disease risk. Nicotine replacement therapy and bupropion are effective treatments for smoking cessation and are most effective when combined with behavioral counseling. Intensive multicomponent interventions that include a case-management component have produced the highest smoking cessation rates for patients who suffer a myocardial infarction. PMID- 10685129 TI - Cardiovascular disease in diabetic patients. AB - Cardiovascular disease is the most frequent cause of morbidity and mortality in patients with diabetes. This article evaluates the results of clinical interventions that have been tested for their ability to ameliorate cardiovascular disease through identification and management of cardiovascular risk factors. These include hyperglycemia, insulin resistance, dyslipidemia, hypertension, cigarette smoking, and a procoagulant state. Attention to all cardiovascular risk factors is necessary if physicians are to reduce the burden of these complications in diabetic patients. PMID- 10685130 TI - Effects of dietary modification and treatment of obesity. Emphasis on improving vascular outcomes. AB - This article has considered a vast literature attesting to the efficacy of dietary intervention on risk factors and on vascular outcomes. Rather than rely solely on pharmacotherapy to improve risk factors and vascular outcomes, physicians, nurses, dietitians, pharmacists, and medical providers should emphasize the benefits of a well-balanced, nutritionally sound dietary program. It should be low in SFA; controlled in calories to avoid (or reduce) obesity; and rich in fruits, vegetables, whole-grain products, and good sources of protein. Emphasis on foods rich in n-3 fatty acids shows promise for reducing cardiovascular outcomes. Further studies using these and antioxidants are eagerly awaited. PMID- 10685131 TI - Chronic infection and coronary artery disease. AB - On a variety of fronts, chronic infection has been found to be significantly associated with the development of atherosclerosis and the clinical complications of unstable angina, myocardial infarction, and stroke. For the most part, these are still just associations. Specific causative relationships on par with that determined between H. pylori and peptic ulcer disease have not yet been established. Potential mechanisms whereby chronic infections may play a role in atherogenesis are myriad. In the case of C. pneumoniae, the effect may result from direct vessel wall colonization, which may damage the vessel directly or indirectly by initiating immunologic responses. In other cases, the effect may simply be that of enhancing the preexisting chronic inflammatory response of the body to standard risk factors, such as hyperlipidemia. Even though the infectious agent may not directly infect the vessel wall, it may perform its critical role from afar. Chronic infection might also influence preexisting plaque by enhancing T cell activation or other inflammatory responses that may participate in the destabilization of the intimal cap. Chronic infection may play a role in the initiation, progression, or destabilization of atherosclerotic plaques. The infectious agents with the most evidence to support a causative role in atherosclerosis include C. pneumoniae and cytomegalovirus. Evidence is mounting for a variety of other potential agents, including H. pylori, various periodontal agents, and even hepatitis A. Future studies are expected to elucidate further the pathophysiologic relationship between chronic infection and atherosclerosis and to evaluate the potential of a variety of treatment approaches, including antibiotics. PMID- 10685132 TI - C-reactive protein, inflammation, and coronary risk. AB - Data have revealed interactions between baseline concentration of hs-CRP and the efficacy of common pharmacologic therapies in primary and secondary prevention, suggesting not only that it may be possible to modify the increased risk associated with elevated hs-CRP, but also that inflammatory markers may be useful in targeting preventive therapies. Inflammatory markers may become a valuable component of routine cardiovascular risk assessment. PMID- 10685133 TI - Antiplatelet and anticoagulant therapy in the secondary prevention of ischemic heart disease. AB - Antiplatelet and anticoagulant medications play a major role in the secondary prevention of ischemic heart disease. Numerous trials have demonstrated their clinical benefits. Newer agents, such as clopidogrel, have challenged aspirin's role as the premier medication for secondary prevention. Much remains to be learned, however, about the merits of these different drug classes, relative to one another and in combination. PMID- 10685134 TI - Hormone replacement therapy and coronary heart disease. A new debate. AB - Hormone replacement therapy is extensively used in the United States, especially for the possible prevention of heart disease, osteoporotic fractures, and, more recently, dementia. The results of recent clinical trials have raised new questions about the risk and benefits of estrogen or estrogen/progesterone therapy to prevent heart attacks, the choice of specific drug therapy such as specialized estrogen receptor modulation (SERM). The change in risk factors, especially weight gain or obesity, may determine the risks and benefits of hormone replacement therapy. PMID- 10685135 TI - Prospects for genetic therapy of cardiovascular disease. AB - Significant progress has been made in cardiovascular gene therapy. Further investigations are required to understand the basic science of vectors, mechanisms of gene delivery, vector-associated immunogenicity, and pathophysiology of vascular and myocardial diseases. In addition, catheter and stent devices will be required to deliver vectors to the vasculature and myocardium (Fig. 3). Despite these scientific challenges, molecular therapies for cardiovascular diseases are being implemented into clinical practice. PMID- 10685136 TI - Homocysteine, B vitamins, and coronary artery disease. AB - This article discusses the metabolism of homocysteine, factors affecting its plasma level, and the evidence for its role in the pathogenesis of vascular disease. The treatment of hyperhomocysteinemia and its possible impact on vascular disease prevention and progression are described also. PMID- 10685137 TI - Antioxidants for vascular disease. AB - The epidemiologic data do not support a strong role for vitamin C in reducing risk of coronary disease. The evidence supporting a protective effect for the family of dietary carotenoids is stronger, but any important protective effect attributable to the specific supplementation of beta-carotene can be excluded. Conversely, results from observational and experimental studies consistently support an effect of vitamin E supplementation on reducing risk of coronary heart disease. The evidence suggests that the major effect, if any, is found at supplemental intake levels at or greater than 100 IU/d. If confirmed in further trials, the net benefit of vitamin E supplementation among populations with existing coronary disease may be substantial, although the current available evidence is insufficient to warrant a change in public policy recommendations. PMID- 10685138 TI - Effects of exercise and cardiac rehabilitation on cardiovascular outcomes. AB - Comprehensive cardiac rehabilitation for coronary patients includes a systematic approach to the measurement and treatment of coronary risk factors, along with the better-known exercise training component. Studies of exercise and nutritional interventions in patients with coronary heart disease have documented improved primary outcomes of decreased morbidity and mortality, decreased symptoms, and fewer cardiac rehospitalizations. Quality of life, depression scores, and physical functioning are improved after rehabilitation. PMID- 10685139 TI - Putting prevention into daily practice. AB - The future holds promise for expanding preventive therapies to reduce cardiovascular risk. These preventive concepts are supported by sound science and strong evidence from multiple randomized clinical trials. To be successful, the health care system must continue to provide financial resources to support physicians and other health care providers in preventive cardiovascular efforts. In the long run, this support should result in a decrease in the need for expensive high-technology, acute-care interventions. Health care provider teams involving physicians, nurses, and other providers will be necessary to ensure the success of these measures, and they must be integrated into the expanding network of inpatient and outpatient delivery systems. Finally, programs to train fellows, residents, and medical students in preventive skills will provide the basis for expanded application of cardiovascular risk therapies and contribute to the ultimate widespread success in decreasing the morbidity and mortality from cardiovascular disease. PMID- 10685140 TI - A concise review of the cost-effectiveness of coronary heart disease prevention. AB - Coronary heart disease is one of the largest sources of morbidity, mortality, and health care expenditure in the United States. This article reviews a number of studies that estimate the cost per unit of health benefits associated with different primary and secondary prevention strategies for coronary heart disease. Although prevention does not provide a panacea for rising health care spending, many preventive strategies are cost-effective when compared to other common clinical interventions. Prevention should be incorporated into regular clinical practice. PMID- 10685141 TI - Embryology, anatomy, and surgical applications of the preperitoneal space. AB - The preperitoneal space is presented from an embryologic, anatomic, and surgical standpoint in detail. Because this space is one of the most used areas for the repair of groin hernias, knowledge of its embryology and anatomy is essential. PMID- 10685142 TI - Transversalis fascia rediscovered. AB - The transversalis fascia is a layer in the make-up of the posterior inguinal wall. It is the deepest, thinnest, and least important layer in terms of the prevention of herniation. It is a segment of the wider endoabdominal fascia. The true posterior wall of the inguinal canal is formed, in varying degrees, by the muscles or aponeuroses of the internal oblique and transversus abdominis. Plainly, Daedalus was not needed to show surgeons and anatomists how to make a labyrinth out of a rabbit hole! PMID- 10685143 TI - The inguinofemoral area from a laparoscopic standpoint. History, anatomy, and surgical applications. AB - No significant difference has been found between early and new diagrams of the posterior anatomy of the inguinofemoral area from a laparoscopic standpoint because anatomy is unique to each individual. But new dangers can arise from new approaches, even if the anatomic structures are well known, so anatomic research is still useful. It provides, relative to new surgical techniques, new vision of structures known for centuries. PMID- 10685144 TI - The inguinal rings. AB - The inguinofemoral area constitutes the frontier between the abdomen and the lower limb. Because of the human standing position, the inguinal region is a zone supporting the abdominal thrust, and is weakened by the orifice of the inguinal and femoral passages. Peritoneal diverticula may externalize into these orifices, leading to the formation of hernias. This article reviews the anatomic constituents of the inguinofemoral region and the anatomic basis for the treatment of hernias. PMID- 10685145 TI - Obturator hernia. Embryology, anatomy, and surgical applications. AB - Obturator hernia is a rare clinical entity. In most cases, it produces small bowel obstruction with high morbidity and mortality. The embryology, anatomy, clinical picture, diagnosis, and surgery are presented in detail. PMID- 10685146 TI - Embryologic and anatomic basis of esophageal surgery. AB - The embryogenesis, congenital anomalies, and surgical anatomy and applications of the esophagus for benign and malignant processes are detailed in this article. Emphasis is placed on the role of embryology and the anatomy involved in surgical decisions. PMID- 10685147 TI - Embryology and surgical anatomy of the mediastinum with clinical implications. AB - This article discusses general mediastinal embryology, and provides anatomy and algorithms for the investigation of mediastinal masses. The superior, anterior, middle, and posterior mediastina also are detailed. PMID- 10685148 TI - Embryologic and anatomic basis of duodenal surgery. AB - The following points should be remembered by surgeons (Table 1). In writing about the head of the pancreas, the common bile duct, and the duodenum in 1979, the authors stated that Embryologically, anatomically and surgically these three entities form an inseparable unit. Their relations and blood supply make it impossible for the surgeon to remove completely the head of the pancreas without removing the duodenum and the distal part of the common bile duct. Here embryology and anatomy conspire to produce some of the most difficult surgery of the abdominal cavity. The only alternative procedure, the so-called 95% pancreatectomy, leaves a rim of pancreas along the medial border of the duodenum to preserve the duodenal blood supply. The authors had several conversations with Child, one of the pioneers of this procedure, whose constant message was to always be careful with the blood supply of the duodenum (personal communication, 1970). Beger et al popularized duodenum-preserving resection of the pancreatic head, emphasizing preservation of endocrine pancreatic function. They reported that ampullectomy (removal of the papilla and ampulla of Vater) carries a mortality rate of less than 0.4% and a morbidity rate of less than 10.0%. Surgeons should not ligate the superior and inferior pancreaticoduodenal arteries because such ligation may cause necrosis of the head of the pancreas and of much of the duodenum. The accessory pancreatic duct of Santorini passes under the gastrointestinal artery. For safety, surgeons should ligate the artery away from the anterior medial duodenal wall, where the papilla is located, thereby avoiding injury to or ligation of the duct. "Water under the bridge" applies not only to the relationship of the uterine artery and ureter but also to the gastroduodenal artery and the accessory pancreatic duct. In 10% of cases, the duct of Santorini is the only duct draining the pancreas, so ligation of the gastroduodenal artery with accidental inclusion of the duct is catastrophic. With the Kocher maneuver, surgeons reconstruct the primitive mesoduodenum and achieve mobilization of the duodenum, which is useful for some surgical procedures. Surgeons should not skeletonize more than 2 cm of the first part of the duodenum. If more than 2 cm of skeletonization is done, a duodenostomy using a Foley catheter may be necessary to avoid blow-up of the stump secondary to poor blood supply. Proximal duodenojejunostomy is advised for the safe management of patients with difficult duodenal stumps. Roux-en-Y choledochojejunostomy and duodenojejunostomy divert bile and food in the treatment of the complicated duodenal diverticulum. The suspensory ligament may be transected with impunity. It should be ligated before being sectioned so that bleeding from small vessels contained within can be avoided. Failure to sever the suspensory muscle completely, which is possible if the insertion is multiple, fails to relieve the symptoms of vascular compression of the duodenum (Fig. 18). Mobilization, resection, and end-to-end anastomosis of the duodenal flexure have been performed as a uniform surgical procedure, avoiding the conventional gastrojejunostomy. With a large, penetrating posterior duodenal or pyloric ulcer, surgeons should remember that The proximal duodenum shortens because of the inflammatory process (duodenal shortening) The anatomic topography of the distal common bile duct and the opening of the duct of Santorini and the ampulla of Vater is distorted Leaving the ulcer in situ is wise Careful palpation for or visualization of the location of the ampulla of Vater or common bile duct exploration with a catheter insertion into the common bile duct and the duodenum are useful procedures In most cases, the common bile duct is located to the right of the gastroduodenal artery at the posterior wall of the first part of the duodenum. (ABSTRACT TRUNCATED) PMID- 10685149 TI - Ampulla of Vater. Anatomic, embryologic, and surgical aspects. AB - The region of the ampulla of Vater constitutes a complex anatomic and functional entity, the biliopancreaticoduodenal confluence, of which the essentials of this rapid review are the: Variation in site of implantation of the greater duodenal papilla, whereas the relations between the common bile duct and the main pancreatic duct are relatively constant Presence at this site of a weak point in the duodenal wall, commonly the site of mucosal diverticula Interdependence of the parietal duodenal mucosa and the sphincteric system of Oddi Existence of an extramural zone of this sphincter, which should be the only one involved in sphincterotomy Danger of wide excisions of the papilla, which, apart from the risk for hemorrhage, cause a breach of the digestive barrier The ampulla of Vater corresponds to the dilated junction of the common bile duct and main pancreatic duct, if present. The ampulla is an extensive anatomic and functional region that includes not only the choledochopancreatic junction but also the sphincter of Oddi, the whole traversing the duodenal wall to open at the greater duodenal papilla. The chief anatomic features of this biliopancreaticoduodenal junction have been reviewed, forming the basis of techniques of surgical or endoscopic sphincterotomies and localized excisions of vaterian tumors. PMID- 10685150 TI - Surgical embryology and anatomy of the diaphragm with surgical applications. AB - This article reviews the development, surgical anatomy, and teratology of the diaphragm, and discusses the diagnostic procedures, surgical therapy, and prognosis of congenital disturbances. Special attention is paid to the traumatic rupture of the diaphragm, concerning incidence, cause, diagnosis, prognosis, and surgical repair. PMID- 10685151 TI - Functional anatomy of the gastroesophageal junction. AB - The study of the functional anatomy of the gastroesophageal junction allows for the demonstration of a double mechanism that combats the conflict of pressures that tends to lead to gastroesophageal reflux. On one hand, the LES, an intrinsic structure, is directly related to the muscle fibers of the organ and responds to a neurohormonal physiologic command. On the other hand is an anatomic entity, centered by the crura of the diaphragm, closely related to the movements of respiration. These structures constitute a second, extrinsic sphincter that gives rise to the zone of high pressure in the terminal esophagus. This role is difficult to assess, and its importance is underestimated. The proper functioning of these two mechanisms implies that the gastroesophageal junction remains in place within the diaphragmatic channel of the esophagus. Also important are the postural phenomena associated with the sloping position of the fundus. In patients with gastroesophageal reflux, the decrease of the pressure measured in the terminal esophagus accounts for the occurrence of reflux. Investigators concede that, under the influence of abdominal straining, the gastroesophageal junction tends to ascend into the diaphragmatic channel. The results are twofold: (1) the muscle fibers of the lower esophagus relax, explaining the incompetence of the intrinsic sphincter, and (2) the sphincteric zone is withdrawn from its muscular diaphragmatic environment. Physicians should consider these structures as a whole in approaching the surgical treatment of reflux. The construction of a periesophageal valve has no anatomophysiologic basis. A gastropexy procedure must be added to replace the gastroesophageal junction in its anatomic setting and keep it there. This procedure also allows retightening of the muscle fibers of the esophageal wall, which is essential in long-term surgical correction. PMID- 10685152 TI - The salivary glands. Embryology, anatomy, and surgical applications. AB - This article discusses the embryology and surgical treatment of diseases of the salivary glands, which require precise knowledge of anatomy. Knowledge of the course of the facial nerve enables safe removal of parotid tumors. The submandibular gland is associated intimately with the lingual and hypoglossal nerves. PMID- 10685153 TI - The greater omentum. Anatomy, embryology, and surgical applications. AB - The significance of the greater omentum has been discovered recently by surgeons of various disciplines because it provides an excellent plastic material against inflammation and irradiation and for repair of defects that can be applied in the abdominal cavity; or it can be exteriorized and lengthened at a vascular pedicle and detached using microvascular anastomoses. Anatomic features, such as the volume of the omentum and the arrangement of the blood vessels, determine the lines of dissection. The eminent omental potential and the different biochemical and immunologic functions are unique, and can be related to specific anatomic structures, some of which may be drawn back to its embryologic sources. The ability of absorption and adhesion formation, neovascularization, and infection defense by the omentum protects against irradiation damage, accelerates healing of dead space, and improves the complication rate and quality of life after application to a wound bed. PMID- 10685154 TI - Appendix and cecum. Embryology, anatomy, and surgical applications. AB - Surgeons should be familiar with surgery of the cecum and appendix because the diseases of this region, especially appendicitis, are the most common indications for surgical exploration. Usually, diagnosis of appendicitis and appendectomy are not difficult, but atypical location of the appendix or other anatomic anomalies can make the diagnosis of appendicitis and appendectomy difficult. In cases of atypical anatomy or diffuse clinical picture, especially in young adults or elderly patients, the spectrum of embryologic and anatomic anomalies must be kept in mind to make the correct treatment decision for individual patients. If doubt persists, explorative laparotomy must be performed to avoid overlooking rare, acute, intra-abdominal abnormalities. PMID- 10685155 TI - Embryology and anatomy of the anorectum. Basis of surgery. AB - The rectum is a pelvic organ, complex in its morphology and its topographic relationships. Its double embryologic origin explains the two types of tumors that develop in the rectum: (1) lieberkuhnian adenocarcinoma in the pelvic rectum and (2) squamous epithelioma in the anal canal. Its venous and lymphatic supply, intensively developed, realizes early pathway of tumoral dissemination. The pelvic relationships of the rectum and anus explain the technical difficulty of rectal surgery, especially when subperitoneal resection and anastomosis are concerned. Imaging of this area permits an early diagnosis of rectal tumors and allows a less invasive surgery with a carcinologic precision. PMID- 10685156 TI - Anatomic basis of hepatic surgery. AB - Modern hepatic surgery is based on precise anatomic foundations. The importance of this information applies to all levels of the diagnostic and therapeutic chain. Modern methods of imaging--CT scanning, MR imaging, and preoperative sonography--help physicians to detect variations and plan surgical excision. PMID- 10685157 TI - Embryology, anatomy, and surgical applications of the extrahepatic biliary system. AB - As technology has improved and the ability to apply this technology in the surgical arena has grown, surgeons have been able to perform more sophisticated operative procedures. Hepatobiliary surgeons are now able to use laparoscopy, immunosuppressive drugs, and technical advances in cryosurgery to accomplish magnificent results. The success and safety of laparoscopic cholecystectomy, orthotopic liver transplantation, and trisegmentectomy for hepatic tumors depend on a high regard for and an accurate knowledge of the anatomy and some of the common embryologic anomalies of the biliary tree. The blood supply, ductal variations, and gallbladder anatomy of this area are often the source of major challenge to unprepared and unaware surgeons. The authors have attempted to stimulate an interest in, a respect for, and perhaps some desire to learn more about the important and fascinating anatomy of this region. PMID- 10685158 TI - Embryology, anatomy, and surgical applications of the kidney and ureter. AB - This article discusses the embryology and anatomy of the kidney and ureter. Surgical approaches, such as the lumbar and thoracoabdominal, are provided. Operations for kidney (i.e., radical nephrectomy, nephroureterectomy, and partial nephrectomy) and ureteric tumors also are discussed. PMID- 10685159 TI - Surgical anatomy and embryology of the adrenal glands. AB - Gross anatomy explains the different surgical approaches to adrenalectomy and the difficulties encountered by surgeons during this procedure. Development of the adrenal glands explains the location of the ectopic sites and excess hormone production by adrenal tumors. The choice of a surgical approach is sometimes difficult and is dependent on (1) the morphology of the body; (2) the volume of the tumor, which necessitates immediate vascular control; and (3) the type of disease, which may necessitate a complete exploration of the abdominal cavity. PMID- 10685160 TI - Embryology, anatomy, and surgical exposure of the great abdominal vessels. AB - This article describes the embryology of the abdominal aorta and the anatomic features of its major visceral branches, including the celiac, superior mesenteric, and inferior mesenteric arteries. The common anatomic variants of these visceral vessels also are reviewed. Various operative techniques to gain surgical exposure to these vessels are described. PMID- 10685161 TI - Veterinary surgeon's guide to Australian bat lyssavirus. AB - Veterinary surgeons in Australia must be aware of the emerging viral diseases and their potential effects on public health generally and, more specifically, on the veterinary profession. Australian bat lyssavirus was identified in 1996 and causes rabies-like disease in bats and humans. Two humans from Queensland have died of Australian bat lyssavirus encephalitis. Surveillance has shown that all Australian bats must be considered carriers of this new virus, therefore protective apparel should be used when handling bats. The pre-exposure regimen of inactivated rabies vaccine (Pasteur Merieux) provides protection against infection. As part of the preventive regimen, at risk groups, such as veterinary surgeons, should seriously consider pre-exposure rabies vaccination. The post exposure protocol involves administration of human rabies immunoglobulins and five intramuscular injections of the inactivated rabies vaccine. PMID- 10685162 TI - Postoperative performance of racing horses with tearing of the medial palmar intercarpal ligament. AB - OBJECTIVE: To examine the relationship between medial palmar intercarpal ligament (MPICL) tearing and postoperative performance in racing horses. MATERIALS AND METHODS: The postoperative performance of 42 horses in which the midcarpal joint was examined arthroscopically was followed prospectively. Intra-articular variables examined were the severity of MPICL tearing, subchondral bone damage and articular cartilage damage. Using a scoring system based on the class of race and the position in the race, a mean score was calculated for up to five races before and after surgery. The preoperative score was subtracted from the postoperative score to give a net score. Statistical analysis was by a Mann Whitney U test and multiple linear regression. RESULTS: Thirty-two (76%) raced postoperatively, 23 (55%) won at least one race and 12 of 26 (46%) performed at the same or higher level. Horses with grades 2 to 4 MPICL tearing had significantly lower net scores than those with grade 1 or no tearing. The severity of subchondral bone damage was the only variable on its own that was significantly correlated with net score (r2 = 0.23, P < 0.05). The addition of the grade of MPICL tearing to bone damage significantly improved the prediction of postoperative performance (P < 0.05). The inclusion of articular cartilage damage had no effect on the prediction of postoperative performance. CONCLUSION: Tears involving more than one third of the MPICL as observed arthroscopically have a significant detrimental affect on postoperative performance of racing horses. PMID- 10685163 TI - Fungal pneumonia in a captive black rhinoceros. AB - A captive black rhinoceros (Diceros bicornis) with a hoof abscess was treated with long-term antibiotic therapy. After 9 months of treatment, there was rapid deterioration, marked weight loss and reluctance to stand. Profuse, bilateral epistaxis developed accompanied by collapse and the animal was euthanased. Necropsy revealed pulmonary aspergillosis with concurrent Pseudomonas aeruginosa infection. Though a well-recognized disease of black rhinoceros, fungal pneumonia has not been reported in this species in Australia. The cost and efficacy of treatment have been questioned, however, prophylactic antifungal drug administration will be considered in any further cases of chronic, debilitating illness in black rhinoceros at Western Plains Zoo. PMID- 10685164 TI - Advantages and disadvantages of using alpha-2 agonists in veterinary practice. PMID- 10685165 TI - Xylazine and medetomidine in small animals: these drugs should be used carefully. PMID- 10685166 TI - Kinetics for clinicians. PMID- 10685167 TI - Babesia and Ehrlichia seropositive horses temporarily imported into Australia. PMID- 10685168 TI - Mortality of sheep exported by sea: evidence of similarity by farm group and of regional differences. AB - OBJECTIVE: To determine whether mortality of sheep exported by sea is similar for sheep from the same farm exported in different years or is associated with the region of origin. DESIGN: Mortalities were monitored in farm groups of sheep exported from the southwest of Western Australia under normal commercial conditions. PROCEDURE: Mortalities were monitored on commercial shipments from 1985 to 1996. For each consignment, the mortality rate was assigned its percentile ranking within the month and year of loading of the ship. A mortality rate was high if its percentile ranking was above a selected cut-off value. Five cut-off values were used in separate analyses. The spatial distribution of farms with high mortality was compared between and within zones of rainfall and length of pasture-growing season. RESULTS: A total of 479 groups of sheep from 405 farms was monitored. Mortality rates ranged from nil to 28.2%. Half of all deaths were from 14.2% of the consignments. There was a significant association (P < 0.05) between the category of mortality (high or low) in the first and second years of monitoring for four of the five cut-off values. The spatial analyses indicated that there were more high-mortality groups, and the average mortality was higher, in the zones of higher rainfall and longer pasture-growing season (P < 0.001). CONCLUSION: Mortality data can be used to identify regions and groups of sheep that are at risk of suffering high death rates when exported by sea. PMID- 10685169 TI - Effect of topical rh-TGF-beta 1 on second intention wound healing in horses. AB - OBJECTIVE: To investigate the effects on wound healing of transforming growth factor-beta 1 as a topical treatment to full-thickness, excisional wounds of the distal limb of horses. DESIGN: A randomised block study using four horses, each with wounds assigned to four treatment groups. ANIMALS: Four adult Standardbred geldings. PROCEDURE: Four, 4 cm2, full-thickness wounds were created on the dorsomedial and dorsolateral aspect of the metacarpus or metatarsus of each limb of four horses, giving a total of 64 wounds. For each limb, wounds were randomly assigned to four treatment groups: no treatment (control), carrier (Methyl Cellulose gel), 50 ng/wound rhTGF-beta 1 in carrier, and 500 ng/wound rhTGF-beta 1 in carrier. Wounds were treated on day 0 and day 8. Effects of treatment were evaluated on the basis of the presence of exuberant granulation tissue requiring excision, number of times excision was required, total wound area, area of epithelialisation, area of granulation, and histological evaluation of biopsy samples of wounds on day 8 and excised wounds on day 21. RESULTS: Topical application of TGF-beta 1 at the two concentrations studied had no significant effect on the total area of wounds (P = 0.7), the area of granulation tissue (P = 0.78), the area of epithelialisation (P = 0.92), histological assessment or subjective clinical assessment of wounds. CONCLUSION: TGF-beta 1 had no beneficial effects on wound healing. Additional trials are needed to test if it has value for wound treatment in horses. PMID- 10685170 TI - Acute cortisol responses of lambs to ring castration and docking after the injection of lignocaine into the scrotal neck or testes at the time of ring application. AB - OBJECTIVE: To test whether injecting lignocaine into the scrotal neck 5 to 10 s before or into both testes immediately after ring castration and docking wound significantly reduce the plasma cortisol response to castration and docking. DESIGN: A physiological study with controls. PROCEDURE: Lambs were given one of six treatments: control handling, injection of lignocaine into scrotal neck, injection of lignocaine into both testes, ring castration and docking, ring castration and docking after lignocaine was injected into the scrotal neck, and ring castration and docking before lignocaine was injected into both testes. Blood samples were taken before and regularly after treatment and analysed for plasma cortisol concentrations. RESULTS: The plasma cortisol concentrations of lambs castrated and docked after lignocaine had been injected into the scrotal neck were significantly lower between 20 and 60 min after treatment than in lambs castrated and docked without local anaesthesia. Injecting lignocaine into the testes after ring application did not significantly reduce the cortisol response to ring castration and docking. CONCLUSIONS: Lignocaine injected into the scrotal neck 5 to 10 s before ring castration will reduce the cortisol response and by inference the pain associated with ring castration. PMID- 10685171 TI - Tail docking and beliefs about the practice in the Victorian dairy industry. AB - OBJECTIVE: To determine the occurrence of tail docking and beliefs about the practice in the Victorian dairy industry. DESIGN: Survey responses were analysed using chi-square tests and by correlation and regression analyses to determine associations between husbandry practices and beliefs and to identify possible predictive variables in relation to docking. PROCEDURE: A survey of the occurrence of docking and beliefs about the practice was conducted in 1997 using face-to-face interviews of 313 respondents at 234 Victorian dairy farms. RESULTS: On average, 35% of dairy farms routinely docked cattle. The practice varied from 11 to 63% in different regions and 12% of stud farms docked their cows. Rubber rings were used on 75% of farms and the average age of the cow at docking was 18 months. Twenty-two percent of cows were docked at a level above the top of the udder and 54% were docked level with the top of the udder. Respondents that docked believed that milking was finished quicker, the risks of leptospirosis for the operator and mastitis for the cow were reduced, the cows were easier to handle, fly numbers were reduced and milk quality was improved. There was a general belief that intact tails could cause significant discomfort to the operator and that docking resulted in acute but not chronic pain. CONCLUSIONS: Docking is an entrenched practice in the Victorian dairy industry. Those farmers who docked generally believed that it was a highly desirable farming practice with particular benefits for the operator. PMID- 10685172 TI - Effect of breed of cattle on innate resistance to infection with Anaplasma marginale transmitted by Boophilus microplus. AB - OBJECTIVE: To assess the innate resistance of and transmission in naive Bos taurus cross Bos indicus and purebred Bos indicus cattle when placed in a paddock with cattle infected with Anaplasma marginale and carrying Boophilus microplus ticks. DESIGN: A group of 49 purebred B indicus, and 48 B indicus cross B taurus (50%, F1 generation) 24-month-old steers were kept in the same paddock with cattle artificially infected with a virulent isolate of A marginale and Boophilus microplus. The cattle were seronegative for A marginale at the start of the trial but had previously been exposed to Babesia bovis and B bigemina. PROCEDURE: Cattle were inspected twice weekly for 118 days. Whole blood, blood smears and serum samples were collected from the cattle on day 37 after exposure and then at regular intervals to day 83 after exposure to measure packed-cell volumes, parasitaemias and antibody titres to A marginale. Any animals that met preset criteria were treated for anaplasmosis. On day 83 all cattle were treated with an acaricide and cattle infected with A marginale were removed from the rest of the group. RESULTS: A marginale was detected in blood smears from 14 crossbred and 9 B indicus steers between days 56 and 72 after exposure. Five and two of the infected crossbred and B indicus steers required treatment, respectively. One of the Bos indicus cattle died as a result of the A marginale infection despite treatment. Antibodies to A marginale were detected in the 23 infected cattle. The mean packed-cell volume depression was 40 and 37% in the affected crossbred and Bos indicus groups, respectively. There was no significant difference detected in susceptibility between these two groups. CONCLUSIONS: Innate resistance of purebred B indicus and crossbred cattle was not significantly different. The results confirm that purebred B indicus and crossbred cattle are sufficiently susceptible to warrant the use of vaccination against Anaplasma infections. PMID- 10685173 TI - Maxillary deformity in a wild platypus (Ornithorhynchus anatinus). PMID- 10685174 TI - Courts ignore reality. PMID- 10685175 TI - Tail docking lies. PMID- 10685176 TI - Microchipping--why worry? PMID- 10685177 TI - EU beef wars expose sewage sludge scandal. PMID- 10685178 TI - Cutaneous phycomycosis in two horses. PMID- 10685179 TI - Anorexia and an abdominal mass in a cat. PMID- 10685180 TI - Ulceration and stricture of the right dorsal colon after phenylbutazone administration in four horses. AB - Four cases of ulceration and stricture of the right dorsal colon were encountered. Ulceration of the right dorsal colon is generally associated with nonsteroidal anti-inflammatory drug (NSAID) toxicosis but there are few reports of stricture following ulceration. All four horses had recent phenylbutazone use: three had been given doses well in excess of the recommended dose and in one the dose was marginally above those recommended but was combined with administration of other NSAIDs. All four horses presented with intermittent low-grade colic, weight loss and ventral oedema. Diarrhoea was also seen in three of them. All had hypoproteinaemia due to severe hypoalbuminaemia, and hyperfibrinogenaemia. Hypoalbuminaemia was less severe in one horse and this horse was successfully managed medically. Two cases were definitively diagnosed at exploratory celiotomy and two at necropsy. Exploratory celiotomy was performed in two horses: one was euthanased at surgery and one was managed successfully with medical treatment and remained normal 1 year after surgery. Medical management included feeding of a low-roughage pelleted ration, corn oil, psyllium mucilloid, and discontinuation of NSAID administration. PMID- 10685181 TI - The distribution and identification of dangerously venomous Australian terrestrial snakes. AB - The identification of dangerous Australian snakes is important in instituting therapy for envenomation. Despite the availability of a number of identification guides with varying degrees of generality, identification can be problematic for several reasons. These include a diversity of common names, many of which are inappropriate or regionally applied to different species, identification keys that focus on variable features, intraspecific variation and interspecific convergence in colouration, and recent changes in scientific nomenclature of species and genera. Geographic distribution of the dangerously venomous species can be a useful aid to identification, by limiting the range of options in a region. However, delineation of the limits of distribution relies on fine scale mapping beyond the resolution of most identification guides. This article provides a summary of the geographic limits of the dangerously venomous Australian snakes, with particular emphasis on major population centres, and clarifies some problems in identification, particularly among brown-coloured snakes. PMID- 10685182 TI - Cytology and histopathology of canine leproid granuloma syndrome. AB - OBJECTIVE: To describe the histopathology of canine leproid granuloma syndrome. DESIGN: Histological examination of biopsy specimens taken from dogs with leproid granuloma syndrome. Biopsies were acquired from four veterinary pathology practices after initial examination showed acid-fast bacilli or lesions suggestive of a mycobacterial aetiology. Tissue from 30 cases formed part of a retrospective survey while a further 7 cases were obtained prospectively. PROCEDURE: Tissue samples were fixed in formalin and paraffin embedded. Sections were stained with haematoxylin and eosin and Ziehl-Neelsen stains. Slides were evaluated for the type of inflammatory response and the number of bacteria. In a few cases smears made from crush preparations and needle aspirates were stained with DiffQuik and acid-fast stains. RESULTS: The common cytological feature seen in DiffQuik stained smears was numerous, often spindle-shaped, macrophages with variable numbers of lymphocytes and plasma cells and lower numbers of neutrophils. Usually few to moderate numbers of medium length bacilli were detected within macrophages or extracellularly. Histologically, lesions were chiefly pyogranulomatous within the subcutis and dermis. Pyogranulomas were composed chiefly of epithelioid macrophages and neutrophils but plasma cells and small lymphocytes were scattered throughout the lesions in which giant cells were seen. Lesions were pyogranulomatous in 36 cases and granulomatous with small suppurative foci in one. The numbers of acid-fast bacilli present were very low to low in 22 cases, moderate in 6 and regionally numerous to numerous in 9. Bacteria were pleomorphic, ranging from long, slender filaments in parallel sheaves to short and variably-beaded bacilli or highly beaded to coccoid organisms. The morphology was more uniform in DiffQuik stained smears than in fixed tissue sections. CONCLUSION: The pathology of canine leproid granuloma syndrome is highly uniform and is suggestive of saprophytic mycobacterial involvement. The morphological diversity of acid-fast bacilli probably results from differences in staining characteristics rather than indicating different species of Mycobacterium. While approximately half of the cases had only few organisms present, the veterinary practitioner using a Romanovsky stain such as DiffQuik on aspirated material might expect to obtain a rapid diagnosis in many cases. This would allow differentiation of the syndrome from neoplastic and other diseases of the skin and direct appropriate treatment at the initial presentation. PMID- 10685183 TI - Extent and economic effect of heat loads on dairy cattle production in Australia. AB - OBJECTIVE: To investigate the extent of heat load problems, caused by the combination of excessive temperature and humidity, in Holstein-Friesian cows in Australia. Also, to outline how milk production losses and consequent costs from this can be estimated and minimised. PROCEDURES: Long-term meteorological data for Australia were analysed to determine the distribution of hot conditions over space and time. Fifteen dairy production regions were identified for higher resolution data analysis. Both the raw meteorological data and their integration into a temperature-humidity thermal index were compiled onto a computer program. This mapping software displays the distribution of climatic patterns, both Australia-wide and within the selected dairying regions. Graphical displays of the variation in historical records for 200 locations in the 15 dairying regions are also available. As a separate study, production data from research stations, on-farm trials and milk factory records were statistically analysed and correlated with the climatic indices, to estimate production losses due to hot conditions. RESULTS: Both milk yields and milk constituents declined with increases in the temperature-humidity index. The onset and rate of this decline are dependent on a number of factors, including location, level of production, adaptation, and management regime. These results have been integrated into a farm level economic analysis for managers of dairy properties. CONCLUSION: By considering the historical patterns of hot conditions over time and space, along with expected production losses, managers of dairy farms can now conduct an economic evaluation of investment strategies to alleviate heat loads. These strategies include the provision of sprinklers, shade structures, or combinations of these. PMID- 10685184 TI - Haematological, serum biochemical and serological features of platypuses with and without mycotic granulomatous dermatitis. AB - OBJECTIVE: To determine whether there are haematological, serum biochemical and serological differences between platypuses (Ornithorhynchus anatinus) with and without granulomatous dermatitis due to Mucor amphibiorum infection. An additional objective was to establish reference haematological and serum biochemical ranges for the species in Tasmania. DESIGN: A clinicopathological and serological study. ANIMALS: A total of 37 free-living adult platypuses captured from streams and dams in Northern Tasmania were used in the clinicopathological study. Twenty-seven were clinically normal and 10 had mycotic granulomatous dermatitis. A total of 22 platypuses (20 adult and 2 juvenile) were used for the serosurvey. Eighteen were captured from streams in Northern Tasmania, and four were submitted for necropsy. RESULTS: Platypuses with mycotic ulcerative dermatitis had significantly smaller packed red cell volumes, haemoglobin concentrations, lymphocyte counts, serum cholesterol and calcium concentrations, and higher serum globulin and potassium concentrations than clinically normal animals. The lymphopenia and hyperkalaemia were thought to be clinically significant. Numbers of Trypanosoma binneyi in blood smears were similar between the two groups. Diseased platypuses had higher concentrations of serum antibody against Mucor amphibiorum as determined by ELISA compared to clinically normal platypuses. CONCLUSION: Platypuses affected by mycotic granulomatous dermatitis showed haematological and serum biochemical changes when compared to clinically normal animals from the same Tasmanian sites. A serological survey may be a useful method for detecting the prevalence of exposure to Mucor amphibiorum and humoral immunity in platypus populations both in Tasmania and the mainland of Australia. PMID- 10685185 TI - Presence of cervical vertebral malformation in Dobermann puppies and the effects of diet and growth rate. AB - OBJECTIVE: To investigate the relationship between diet, growth rate and the presence of caudal cervical malformation in Doberman puppies. DESIGN: A prospective study of 15 Dobermann puppies from three unrelated litters, aged from 0 to 16 weeks. PROCEDURE: The growth rate in terms of body weight gain and increase in ulna length were measured weekly for all puppies. In addition the nutritional quality of the diets was assessed. Radiographs of the cervical spine were taken at 6 and between 12 and 16 weeks of age and examined for the presence of caudal cervical vertebral malformation. A mixed model for repeated measures data was used to investigate the relationship between the growth rate of the puppies and the fixed effects age, dam, diet, gender and presence of caudal cervical malformation. RESULTS: Five of the puppies had changes consistent with caudal cervical malformation. The diets fed were either balanced or transiently deficient in protein, calcium, phosphorus and/or magnesium. There was no significant association between growth rate and the variables dam, gender and the presence of caudal cervical vertebral malformation. There was no significant association between diet and increase in ulna length, but a trend existed between body weight gain and the feeding of a balanced diet (P = 0.0672). CONCLUSION: Caudal cervical vertebral changes can be detected radiographically as early as 6 weeks of age in some Dobermann puppies. A balanced diet and growth rate are not significant factors in its initial development. PMID- 10685186 TI - Listeria in zoo animals and rivers. PMID- 10685187 TI - Osmotic fragility of erythrocytes in periparturient yaks. PMID- 10685188 TI - Food safety a major AFFA priority. PMID- 10685189 TI - Giving evidence in court appearances. PMID- 10685190 TI - Draft statement on registration to practise as a veterinarian. PMID- 10685191 TI - Take a stand AVA. PMID- 10685192 TI - Hounding PetPEP. PMID- 10685193 TI - Comprehensive perinatal centers in the era of managed care. PMID- 10685195 TI - Serum cortisol levels in very low birth weight infants of substance-abusing mothers. AB - OBJECTIVE: Recently, hypoadrenalism was described in extremely low birth weight infants. We have been measuring serum cortisol levels in very low birth weight (VLBW) infants to assess their adrenal function. It was noticed that infants of substance-abusing mothers (ISAM) had unusually high serum cortisol levels. To our knowledge, there are no data regarding serum cortisol levels in VLBW ISAM. The objective of our study was to determine if serum cortisol levels of VLBW ISAM are different from serum cortisol levels in VLBW infants without a history of maternal substance abuse. METHODS: We reviewed medical records of infants with birth weight between 400 and 1300 gm who were born between July 1994 and June 1995. Seven ISAM who had serum cortisol levels drawn before 48 hours of life were identified. Seven infants matched for gestational age, antenatal steroid exposure, and birth weight served as control infants. Clinical characteristics, serum cortisol levels, and clinical outcome were compared between the two groups. Serum cortisol levels of all infants were measured by radioimmunoassay. RESULTS: The mean birth weight of the ISAM group was 834 gm (range 480 to 1175 gm), and the mean gestational age was 27.4 weeks. The mean birth weight of the control group was 921 gm (range 525 to 1230 gm) with a mean gestational age of 27.8 weeks. The mean serum cortisol level of the ISAM group was 65.3 micrograms/dl (range 11.9 to 144 micrograms/dl); it was significantly higher than that of the control group (mean cortisol level of 20.5 micrograms/dl; range 4.9 to 62.7 micrograms/dl) (p = 0.01, two-tailed). CONCLUSION: Our data suggest that maternal substance abuse significantly increases serum cortisol levels in VLBW infants. Studies in larger scale are needed to confirm our findings and to correlate cortisol levels in this group of patients with clinical outcome. PMID- 10685194 TI - The New Hampshire Perinatal Program: twenty years of perinatal outreach education. AB - OBJECTIVE: To describe 20 years of regional outreach education by the New Hampshire Perinatal Program, its interaction with all 26 community hospitals in the state with maternity services and an additional four in adjoining Vermont. STUDY DESIGN: This paper describes educational initiatives responsive to the needs of perinatal physicians and nurses. The core of the program is the transport conference held annually at each referring hospital in which maternal fetal and infant referrals are discussed. There are additional community hospital based programs, programs at convenient locations in the region and medical center conferences and skills programs. RESULTS: The program annually awards 10,000 continuing medical education credits (CME) and nursing contact hours. Evaluation and feedback from all participants is encouraged. New Hampshire has one of the lowest perinatal mortality rates in the county, which reflects in part the accomplishments of the program. CONCLUSION: Perinatal outreach education is a shared responsibility of providers in both the academic center and community hospitals and is necessary to ensure optimal care for mothers and infants. PMID- 10685196 TI - Rates of successful vaginal delivery after cesarean for patients with private versus public insurance. AB - OBJECTIVE: To examine the effect of insurance status on method of delivery while controlling for clinical and sociodemographic characteristics of women delivering at a single medical center. STUDY DESIGN: Sociodemographic and clinical characteristics of 878 women, who delivered their infants between 1985 and 1991 at a university hospital, were analyzed. Women were included if their previous infant was delivered by cesarean section and if the current pregnancy was > or = 37 weeks' gestation at nonemergent delivery with insurance status clearly specified. The outcome of interest was the rate of successful vaginal births after cesarean (VBAC) delivery by insurance status. RESULTS: The trial of labor rate for the cohort, defined as the rate of VBAC or cesarean deliveries following labor, was 55%; 61% were vaginal deliveries. Significant differences with regard to insurance status and several maternal factors were noted between trial of labor and cesarean section--no labor groups. After controlling for potentially confounding variables, the Medicaid/indigent group was more likely than the privately insured group to undergo a trial of labor (odds ratio, 1.5; 95% confidence interval, 1.1 to 2.4). Of women who underwent a trial of labor, after controlling for other characteristics, the Medicaid/indigent group was more likely than the privately insured group to deliver vaginally (odds ratio, 1.9; 95% confidence interval, 1.1 to 3.2). CONCLUSION: After controlling for other covariables, women with a history of a prior cesarean section with Medicaid/indigent insurance were more likely than privately insured women to attempt a trial of labor, and subsequently, to deliver vaginally, given that a trial of labor was attempted. PMID- 10685197 TI - Readmission for group B streptococci or Escherichia coli infection among full term, singleton, vaginally delivered neonates after early discharge from Florida hospitals for births from 1992 through 1994. AB - BACKGROUND: In Florida during the period 1992 through 1994, there was a major drop in the length of stay for full-term, singleton, vaginally delivered newborn babies in the hospital. A major concern on the part of clinicians has been the potential of an increased risk of sepsis (manifesting itself after discharge) associated with earlier newborn discharge from the hospital. We used the Florida hospital discharge dataset to study the frequency of readmission with sepsis after early newborn discharge to home. METHODS: Using the Florida Agency for Health-Care Administration Acute Care Hospital Discharge Dataset, we used a multivariate, probabilistic matching algorithm to merge newborn discharge records for births from 1992 through 1994 with readmission discharge records (including hospital-to-hospital transfers and multiple readmissions) during the first 28 days of life. We used the resulting merged dataset to study bacterial infection diagnoses on newborn and readmission records and to examine relationships between readmission diagnoses and timing of newborn discharge among the 364,528 full term, singleton, vaginally delivered babies (FTSVDBs) without congenital anomalies from 1992 through 1994 in Florida acute-care hospitals. RESULTS: Overall, 86.3% of all FTSVDBs born in Florida acute-care hospitals were discharged between day of life (DOL) 0 (born and discharged the same day) and DOL 2 (discharged two calendar days after birth). The group B streptococci (GBS) infection code was found on the newborn discharge records of 9.2 per 10,000 FTSVDBs over the 3-year period, 5.9% of which involved hospital-to-hospital transfer of the baby. Escherichia coli infection codes were found on the records of 3.4 per 10,000 FTSVDBs over the 3-year period, 2.3% of which involved hospital to-hospital transfer of the baby. Of those FTSVDBs discharged to home without infection codes during DOL 1 to 2, 0.8 per 10,000 were readmitted within 7 days (inclusive) with GBS infection, and 2.0 per 10,000 with E. coli infection. When the data for readmitted babies were pooled for 1992 through 1994, the odds ratio for probability of readmission comparing discharges on DOL 1 to DOL 2 for GBS was 2.27 (95% confidence interval, 1.83 to 2.70), and for E. coli 2.16 (95% confidence interval, 1.46 to 2.85). Over this 3-year period, for babies discharged on DOL 1, there was a 115% increase in the rate of readmission for GBS from 1992 through 1994 (p < 0.01) and a 36.5% increase in the rate of readmission for E. coli (p < 0.05). However, among FTSVDBs discharged on DOL 2, the rate of readmission for both GBS and E. coli did not change during the period 1992 through 1994. There were no deaths among FTSVDBs as either newborns, transfers, or readmissions within 7 days of discharge, with either GBS or E. coli infection codes on their discharge record. CONCLUSION: From 1992 through 1994, the increased number of babies discharged early in Florida was temporally associated with an increased rate of readmission during the week after discharge for both GBS and E. coli infection among babies discharged on the calendar day after birth. With an increase in both the number of babies exposed to the risks of early discharge, and an increased rate of these serious infections during the week after discharge from the hospital, the number of these babies grew to exceed, by several fold, the number of babies with inborn errors of metabolism picked up by state screening programs. PMID- 10685198 TI - Use of delayed pushing with epidural anesthesia: findings from a randomized, controlled trial. AB - OBJECTIVE: To compare outcomes between women receiving epidural anesthesia assigned to a group following either a 1-hour "delayed" pushing protocol or directed to initiate pushing at full cervical dilation. STUDY DESIGN: Using a randomized, controlled design, multivariate analyses were used to evaluate second stage labor duration and Apgar scores. An estimated odds ratio equation evaluated fetal descent progress. RESULTS: A 13.68-minute difference occurred in second stage labor length (p = 0.225). No differences were found in Apgar scores (p > 0.09). An estimated odds ratio, that progress in terms of one fetal station unit would occur for control group subjects as compared with subjects with similar progress in the experimental group, was 1.51 (95% confidence interval: 1.16, 1.95). CONCLUSION: Second stage labor was not significantly lengthened, and a similar rate of fetal descent occurred in the absence of directed pushing. Findings support further research on the potential advantages of minimizing the duration of pushing in labor. PMID- 10685199 TI - Skin care practices in the neonatal nursery: a clinical survey. AB - OBJECTIVE: To survey the details of skin care practices in a sample of level I, II, and III nurseries in the United States. DESIGN: A survey conducted by written questionnaire, personal inspection, and phone contact. PARTICIPANTS: Information was obtained from staff physicians and nurses about routine neonatal skin care practices, including bathing, cord care, emollient use, diapering, use of antimicrobial skin preparations, management of intravenous infiltration, approach to diaper rash, and methods used to minimize transcutaneous water loss. SETTING: Fifteen nurseries from twelve hospitals in four states were surveyed. RESULTS: Among the nurseries surveyed, we found no uniform approach to skin care. Only two individual maneuvers were consistently performed in all the nurseries: criteria for bathing and skin antisepsis with povidone-iodine. Other than these, a wide range of practices and products were used, some with a high ratio of risk and/or cost to benefit. CONCLUSION: A better understanding of the principles of infant skin care and a more uniform approach to skin care in the neonatal nursery can minimize risks and costs to this special population of patients. PMID- 10685200 TI - Neonatal seizures: incidence, onset, and etiology by gestational age. AB - OBJECTIVE: To examine the influence of gestational age on seizures in the neonatal intensive care unit. STUDY DESIGN: A cohort of 4165 neonates admitted to a university intensive care unit between 1986 and 1995. The incidence, time of onset, and etiology of neonatal seizures in the cohort were distributed by gestational age. Logistic regression and t test were used to examine the relationship between gestational age and seizures in the neonatal intensive care unit. RESULTS: Seizures occurred in 356 (8.6%) infants. The seizure rate was parabolically related to gestational age, such that infants at 30 to 36 weeks' gestation had a 4.8% rate compared with 11.9% and 14.1% rates for infants < 30 and > 36 weeks, respectively (p < 0.001). Seizures manifested earlier in infants < 30 weeks (2.3 +/- 5.6 days of life) and > 36 weeks (3.7 +/- 8.7 days) gestational age compared with neonates 30 to 36 weeks (10.4 +/- 14.5 days) (p < 0.001). Intraventricular hemorrhage was the principal etiology underlying the higher seizure rate for infants < 30 weeks (p < 0.001). Hypoxic-ischemic encephalopathy and congenital malformations were primary factors for infants > 36 weeks (p < 0.01). Nervous system infections were evenly distributed across gestational age. CONCLUSION: Gestational age exerts a considerable influence on the incidence, onset, and etiology of neonatal seizures. PMID- 10685201 TI - Heart rate and oxygen saturation correlates of infant apnea. AB - OBJECTIVE: To analyze the effects of apnea duration on changes in heart rate and oxygen saturation and to examine the temporal relationships among these variables. STUDY DESIGN: An event analysis sheet was designed to analyze numerous variables reflecting changes in heart rate and oxygen saturation associated with infant apnea. From July 1, 1991 through June 30, 1992 we identified 32 infants enrolled in The Infant Apnea Program at St. Peter's Medical Center, New Brunswick, NJ who had apnea > or = 15 seconds in duration on consecutive 12-hour multichannel recordings of heart rate, thoracic impedance, nasal thermistry, and oxygen saturation. The apnea epochs of these patients were subdivided into apnea of short (10 to 14 seconds), medium (15 to 19 seconds), and long (> or = 20 seconds) duration, and a total of 236 apnea epochs were analyzed. The significance of differences was assessed by analysis of variance and Newman-Keuls multiple comparisons. RESULTS: We found that the duration of apnea has significant effects on perturbations in both heart rate and oxygen saturation, however, the degree of oxygen desaturation can not be predicted by the perturbation in heart rate. Analysis of the temporal relationship of apnea, bradycardia, and oxygen desaturation reveals that, although apnea precedes both heart rate and oxygen saturation drops in most infants as the length of apneic interval increases, the interval between apnea onset and associated drops in heart rate and/or oxygen saturation also increases. CONCLUSION: Oxygen saturation monitoring may provide important physiologic data that can not be assessed by cardiorespiratory monitoring alone. PMID- 10685202 TI - Developmental care teams in the neonatal intensive care unit: survey on current status. AB - Developmental Care Teams (DCT) have evolved in Neonatal Intensive Care Units (NICUs) in response to mounting evidence that developmental care is cost effective and improves outcomes of critically ill newborns. Lack of national practice guidelines and standardized roles for DCT members prompted formulation and distribution of a questionnaire to obtain information regarding staff membership of DCTs, budgeting for DCTs, utilization of developmental care in practice, and education and developmental training of NICU staff. Questionnaires were sent to 50 NICUs in 30 states, with a return rate of 62% (31 of 50), representing 18 different states. Of those who responded, 64% had a DCT, and an additional 24% were in various phases of starting a team. Forty-three percent of the teams meeting on a regular basis did so monthly. Only 30% of those with a DCT had a dedicated budget to cover operating costs of their developmental program. Fifty-two percent of respondents had Neonatal Individualized Developmental Care and Assessment Program (NIDCAP)-certified staff at their institutions; however, nine other types of developmental specialists were also listed. Only four respondents indicated utilization of set criteria for initiation of a DCT consult, and 74% of those with DCTs initiated consults "when the need arises." NIDCAP assessments were used for parent teaching (54%), care plans (69%), care recommendations (46%), and at caregiver "discretion" (39%). The results of the survey validated an intense interest in developmental care. Approach to developmental care is variable between NICUs and implementation as outlined by NIDCAP is unusual. Practical guidelines for utilization and funding of DCTs are needed. PMID- 10685203 TI - The impact of personal problems on accessing prenatal care in low-income urban African American women. AB - OBJECTIVE: The purpose of this study was to investigate the nature and contribution of personal factors related to the use of prenatal care in a sample of high-risk women residing in an urban environment where care was accessible and free. STUDY DESIGN: The sample consisted of 297 African American women with low socioeconomic status and a high school education or less who were newly delivered of neonates. The level of prenatal care was classified according to the Kessner 3 Parameter Index (adequate, intermediate, inadequate). Women who received no prenatal care made up a fourth group. Subjects responded to the "Ten-Item Checklist" of Richwald. Rhodes, and Kersey and an in-person interview that queried their reasons for obtaining different levels of prenatal care. RESULTS: Both personal and structural reasons were described by women for not obtaining care earlier in pregnancy or at all. The mean number of personal and structural problems reported per subject was inversely correlated to the level of prenatal care obtained. However, personal problems were the single most important reason cited by these women. Personal problems that were statistically significant different among the groups were drug use and desire for an abortion. The structural barriers that exhibited statistically significant differences among the groups were trouble scheduling an appointment, access totransportation, dislike of health care professionals and institutions, access to child care, and not knowing where to go. CONCLUSION: Both personal and structural problems were cited as reasons for not obtaining adequate prenatal care. Structural barriers to prenatal care have been identified and extensively studied. These barriers to care continue to persist, despite innovations in program delivery and access. This study demonstrates that the significance of personal problems has not been adequately considered as a major factor associated with insufficient prenatal care. PMID- 10685204 TI - Disseminated coagulopathy associated with transtorcular embolization of vein of Galen aneurysm in a neonate. AB - We describe in this report the development of disseminated intravascular coagulopathy in a neonate after transtorcular embolization of an unusual vein of Galen aneurysm. This rare but potentially fatal complication associated with transtorcular embolization should be considered in decision-making and prognostic evaluation processes, especially in neonates with severe heart failure. PMID- 10685205 TI - Persistent pulmonary hypertension in a neonate with cystic adenomatoid malformation of the lung following lobectomy: survival with prolonged extracorporeal membrane oxygenation therapy. AB - A full-term neonate is reported with congenital cystic adenomatoid malformation of the lung treated by lobectomy with development of pulmonary hypertension. The infant was successfully treated with extracorporeal membrane oxygenation (ECMO) for persistent pulmonary hypertension, which developed postoperatively. An 18-day course of venovenous ECMO was necessary to effectively reverse the severe pulmonary hypertension. This was probably a result of significant pulmonary hypoplasia of the compressed lung. Although not all congenital cystic adenomatoid malformations of the lung are associated with pulmonary hypoplasia and persistent pulmonary hypertension, this is one case where severe pulmonary hypertension developed secondary to a mass effect by a large lesion in the chest. PMID- 10685206 TI - Congenital chylothorax in neonatal thyrotoxicosis. AB - We report a patient with congenital chylothorax who also had neonatal thyrotoxicosis secondary to maternal Graves' disease. Fetal tachycardia with hydrops was detected at 28 weeks' gestational age. The fetus responded to antithyroid medication in utero but had persistent bilateral pleural effusion. At birth, he had respiratory distress due to massive pleural effusion. Cytologic studies of pleural fluid were consistent with chylothorax. To the best of our knowledge, the association of congenital chylothorax with fetal (neonatal) thyrotoxicosis, has not been reported previously. PMID- 10685207 TI - Genetics casebook. Pyloric atresia. PMID- 10685208 TI - Special imaging casebook. Neonatal Bardet-Biedl syndrome with renal anomalies and hydrocolpos with vesicovaginal fistula. PMID- 10685209 TI - Neonatal apnea casebook. Gastroesophageal reflux (GER)-associated apnea. PMID- 10685211 TI - Sedation administered to very low birth weight premature infants PMID- 10685210 TI - Developmentally based care reduces stress levels in severely ill, very low birth weight (VLBW) infants, and thus decreases sedation requirements. PMID- 10685212 TI - Joe Butterfield's panache. PMID- 10685213 TI - An open letter to the Butterfields. PMID- 10685214 TI - L. Joseph Butterfield, MD: a man for all seasons. PMID- 10685215 TI - L. Joseph Butterfield, MD: personal reflections. PMID- 10685216 TI - Newborn country, USA. PMID- 10685217 TI - Reflections about a friend. PMID- 10685218 TI - In honor of Dr. Joseph Butterfield. PMID- 10685219 TI - Personal reflection on Joe Butterfield. PMID- 10685220 TI - The good doctor: a neonatologist's perspective. PMID- 10685221 TI - Parental dreams, dilemmas, and decision-making in cinema verite. AB - Our film Dreams and Dilemmas: Parents and the Practice of Neonatal Care is on its way to meeting its goal of furthering the "Principles for Family Centered Neonatal Care" (Harrison H. Pediatrics 1993;92:643-50) through cinema verite depiction of parental involvement in decision-making. Reality-based filmmaking can provide valuable and successful educational material that advances care and understanding. However, there are real practical and ethical concerns such as privacy, consent, and uncertain or unknown future impact on participants. Successful reality-based filmmaking in a complex medical environment such as a neonatal intensive care unit requires careful attention to ways of ensuring full communication between all those involved and efforts to allay participants' anxiety about being portrayed unfavorably. The most important ingredient, however, is the skill and ability of the filmmaker to engender trust. PMID- 10685222 TI - Very low birth weight births at non-NICU hospitals: the role of sociodemographic, perinatal, and geographic factors. AB - PURPOSE: The purpose of this study was to assess the extent of variation in the percentage of very low birth weight (VLBW) infants born at perinatal Level 1 hospitals (no Neonatal Intensive Care Unit [NICU]) across California's nine geographic Perinatal regions. The role of sociodemographic, perinatal, and geographic factors was also assessed. METHODS: Multivariate analysis of California birth certificate files between 1989 and 1993, for 24,094 live-born infants weighing between 500 and 1499 gm, was conducted to identify factors associated with delivery at a Level 1 hospital. Analyses specific for race and ethnicity were also conducted for Hispanic, African American, and white cohorts. RESULTS: In the 5-year study period, 1989 through 1993, 10.5% (24,094) of all live-born VLBW infants were delivered in Level 1 hospitals. Significant variation across regions was evident, ranging from a regional low of 3.1% to a high of 24.3%. After controlling for multiple factors, the odds of delivering at a Level 1 hospital were decreased for African Americans and South East Asians and increased in Hispanic women as compared with white non-Hispanic women. For all women, less then adequate prenatal care, living in a 50% to 75% urban zip code, and living greater then 25 miles from the nearest NICU significantly increased the odds of VLBW delivery at a Level 1 hospital. For Hispanics, teen pregnancy and having two or more prior infant deaths increased the odds, whereas Medi-Cal as the payer source for delivery and two or more pregnancy complications decreased the odds of a Level 1 VLBW delivery. After taking these factors into account, when compared with Los Angeles, the odds of inappropriate delivery site ranged from 0.37 to 2.75 across California's nine geographic perinatal regions. Of this variation, 78% could be accounted for by the percentage of total births that delivered at a region's Level 1 hospitals. CONCLUSION: The overall state average of 10.5% deliveries of VLBW at Level 1 hospitals, although close to the 10% national objective for the year 2000, did not indicate the wide variation seen across California's nine geographic regions. Risk-adjusted regional differences in the likelihood of inappropriate delivery site for the high-risk VLBW infants suggest that reaching the Healthy People 2000 objective will require further strengthening of California's perinatal regional networks, especially in those regions where a high percentage of total births deliver at Level 1 hospitals. PMID- 10685223 TI - Bilirubin toxicity and differentiation of cultured astrocytes. AB - OBJECTIVE: To study the toxicity of bilirubin in primary cultures of newborn rat cerebral cortical astrocytes. STUDY DESIGN: Primary cultures of newborn rat astrocytes were incubated at bilirubin concentrations of 0, 1, 5, 10, 25, 50, 100, 200, and 2000 microM, at a bilirubin:albumin molar ratio of 1.7. Bilirubin toxicity was determined by changes in cellular morphology, trypan blue staining, and lactate dehydrogenase (LDH) release into the culture medium at various times of incubation. To determine if differentiation of astrocytes affects bilirubin toxicity, cultures were treated with dibutyryl cyclic adenosine monophosphate. RESULTS: All three indices of toxicity showed a bilirubin concentration dependence. LDH release in experimental cultures was significantly elevated (p < 0.05) above that of control cultures by 24 hours at bilirubin concentrations of > or = 100 microM. The absolute amount of LDH release differed significantly between the 200 and 2000 microM cultures from 1.5 to 24 hours, after which duration of exposure appeared to take over and all cultures approached maximum. LDH release for the lower concentrations all reached maximum by 120 hours, except for the 1 microM cultures, which showed no significant elevation above control throughout the study period. At 100 and 200 microM bilirubin, LDH release by untreated cells was significantly higher (p < 0.05) than release by treated cells by 36 hours. CONCLUSION: Undifferentiated astrocytes appeared to be more sensitive to bilirubin toxicity, which may correlate with the greater susceptibility of newborns to kernicteric injury. Studies with primary astrocyte culture may provide insight into how bilirubin sensitivity changes with brain development as well as the cellular and biochemical mechanisms of bilirubin encephalopathy. PMID- 10685224 TI - Outcomes and resource utilization for newborns with major congenital malformations: the initial NICU admission. AB - HYPOTHESIS: Newborns with major congenital malformations (MCM) have contributed to a significant proportion of resource utilization in a regional referral neonatal intensive care unit (NICU). SETTING: The Children's Hospital Medical Center NICU, Cincinnati, OH. SUBJECTS: Newborns with and without MCM admitted from August 1, 1993 through July 31, 1994. Total patients studied were 572; 147 with and 385 without MCM. No intervention was performed in this observational study. STATISTICS: Statistics were t test, chi-squared, and rank sum analysis. RESULTS: MCM accounted for 27.6% of NICU referrals, 32.4% of total NICU days, and 39.6% of NICU costs. Both median cost per patient and length of stay were significantly (p < 0.01) higher for patients with MCM than those without MCM. Surgery was more frequent in MCM than non-MCM cases. Thirty-three percent of the newborns with MCM received ongoing medical support at discharge. CONCLUSION: Patients with MCM remain as one of the largest and costliest groups hospitalized in a referral NICU. PMID- 10685225 TI - Obesity and related pregnancy complications in an inner-city clinic. AB - OBJECTIVE: The study was designed to determine the prevalence of obesity and related pregnancy complications in an inner-city prenatal clinic. STUDY DESIGN: A retrospective review was conducted of the medical records of 281 women with no chronic diseases and who delivered singleton term babies during a 1-year period. The frequencies of various pregnancy complications, including pregnancy-induced hypertension, preeclampsia, gestational diabetes, shoulder dystocia, postpartum hemorrhage, fourth degree laceration, intrauterine growth restriction, and macrosomia, were compared among groups of patients stratified by body mass index (BMI). RESULTS: Thirty-four percent of patients had a reported prepregnancy BMI of > 26 kg/m2. Fifty-two percent of patients were obese (BMI > 26 kg/m2) when they registered for prenatal care, and 82% of patients had a BMI > 26 kg/m2 at the time of delivery. The incidence of birth weights of > 4 kg was significantly higher in women with a registration BMI > 26 kg/m2 (p = 0.026). Most of these macrosomic babies had mothers with a BMI > 29 kg/m2. Patients who required cesarean delivery had significantly higher BMI than those who were delivered vaginally (p < 0.001). CONCLUSION: Obesity was more common in our inner-city population than has previously been reported and was associated with an increased risk of fetal macrosomia and operative delivery. PMID- 10685226 TI - Pulmonary hypertension in children following extracorporeal membrane oxygenation therapy and repair of congenital diaphragmatic hernia. AB - OBJECTIVE: Pulmonary hypertension (PHT) is present in all children at birth, but its degree and rate of resolution in infants diagnosed with congenital diaphragmatic hernia (CDH) requiring extracorporeal membrane oxygenation (ECMO) need to be established. STUDY DESIGN: Twenty-one ECMO/CDH survivors (aged 3.2 +/- 1.4 years) were prospectively evaluated by Doppler echocardiography (ECHO) to determine the presence of PHT. Twenty children without structural heart disease were used as controls. Study patients received a physical examination and an electrocardiograph examination, and their charts were reviewed for neonatal course data. Patients found to have PHT by ECHO received a complete history and exercise treadmill/oxygen desaturation study. RESULTS: Eight of the 21 patients (38%) met echocardiographic criteria for PHT. No neonatal course data were found to be predictive of eventual PHT status. There was no correlation between physical examination or electrocardiographic findings and PHT. Complete histories showed five of the eight patients with PHT had some degree of exercise intolerance and seven had wheezing. Two of the seven patients studied on the treadmill desaturated 5% or greater from baseline. CONCLUSION: There is evidence that PHT either persists or recurs in a significant portion of the ECMO/CDH population and may remain symptomatic well beyond the neonatal period. PMID- 10685227 TI - Massive fetomaternal hemorrhage and fetal death: are they predictable? AB - OBJECTIVE: To report the incidence of massive fetomaternal hemorrhage (FMH) associated with fetal death and to test the hypothesis that FMH is more likely to occur in those with risk factors for FMH. STUDY DESIGN: All cases of fetal death of infants weighing > 500 gm between January 1, 1990 and December 31, 1994 were reviewed for evidence of massive FMH (> or = 2% fetal cells in the maternal circulation as measured by the Betke-Kleihauer test). Women with risk factors were compared with those without risk factors with respect to the occurrence of massive FMH. RESULTS: The prevalence of massive FMH was 14 of 319 (4.4%) cases, occurring in 4 of 102 (3.9%) of those with risk factors and 10 of 217 (4.6%) of patients without risk factors (p = 0.78). Otherwise unexplained fetal death was associated with massive FMH in 5 of 141 (3.5%). Major fetal anomalies were present in 5 of 14 (35.7%) cases of massive FMH. CONCLUSION: Clinical risk factors do not predict an increased likelihood of massive FMH. Massive FMH is associated with fetal anomalies. Betke-Kleihauer testing should be performed in all cases of fetal death, including those with anomalies regardless of the presence or absence of risk factors for FMH. PMID- 10685228 TI - Chemotherapy during pregnancy and its effects on the fetus--neonatal myelosuppression: two case reports. AB - Cancer during pregnancy is infrequent. It presents an ethical dilemma--remission may be obtained with chemotherapy, but it has potential harmful effects to the fetus. We report a case of a very low birth weight preterm female infant born to a 21-year-old mother diagnosed with leukemia and given chemotherapy up to 1 week before delivery. In the laboratory, initial findings included severe pancytopenia, and bone marrow aspiration demonstrated complete aplasia. She was given blood product transfusions, erythropoietin, and granulocyte colony stimulating factor. The hematologic derangement was resolved without documented infections. The second case is a preterm male infant whose 30-year-old mother was diagnosed with lymphoma and had received chemotherapy during the third trimester. The infant presented with moderate leukopenia. He had an uneventful course without documented infection. Exposure of the fetus to transplacental chemotherapy must be considered when evaluating therapy options and timing of delivery in hematologic malignancies diagnosed during pregnancy. PMID- 10685229 TI - Resuscitation of the meconium-stained infant and prevention of meconium aspiration syndrome. PMID- 10685230 TI - Radiology casebook. Gastric perforation with subserosal dissection of air. PMID- 10685231 TI - Asymptomatic intraplacental choriocarcinoma diagnosed on routine placental examination. AB - Asymptomatic intraplacental choriocarcinoma is a rare event with only a few case reports in the literature. The recognition of such a lesion on routine placental examination is important and prompts rapid clinical evaluation for identifying residual and/or metastatic disease followed by institution of chemotherapy. Failure to recognize such a lesion on gross inspection of the placenta can result in disseminated disease, which can be lethal. The following report describes the placental findings and clinical outcome in a recent case with a review of the relevant literature. PMID- 10685232 TI - The California Perinatal Quality Care Collaborative: a model for national perinatal care. PMID- 10685233 TI - Percutaneous central catheters and peripheral intravenous catheters have similar infection rates in very low birth weight infants. AB - OBJECTIVE: We performed this study to determine if percutaneous central lines (PCLs) were associated with infection more often than peripherally placed intravenous catheters (PIVs). STUDY DESIGN: We conducted a retrospective, cohort study of 53 infants with PCLs inserted from March 1993 to February 1995 for evidence of catheter-related bloodstream infection and 97 cohorts with PIVs who were matched to the infants with PCLs by admission date and birth weight. We considered an infant to have catheter-related bloodstream infection if bacteremia occurred while the PCL or PIV was in place with no other identifiable infection focus. Statistical analyses were performed by using either Student's t test or the Mann-Whitney U test where appropriate. RESULTS: There were eight infections per 1000 catheter days of PCL use and nine infections per 1000 catheter days of PIV use. CONCLUSION: PCLs do not become infected more often than PIVs. PMID- 10685234 TI - Fetal erythropoietin levels in growth-restricted and appropriately grown neonates with and without abnormal fetal heart rate tracings: a comparison with cord blood gases and Apgar scores. AB - OBJECTIVE: To determine if umbilical cord plasma erythropoietin (EPO) levels in combination with cord blood gases and Apgar scores can distinguish between subacute and chronic uteroplacental insufficiency. METHODS: A total of 184 neonates delivered between 1993 and 1997 at Tampa General Hospital were studied. Cord plasma EPO levels, cord blood gases, and Apgar scores were determined prospectively and compared in four subgroups that were defined based on the presence or absence of fetal growth restriction (FGR; chronic fetal hypoxia), abnormal fetal heart rate tracings during labor (FHR; subacute/acute fetal hypoxia), or both. RESULTS: Both growth-restricted and appropriately grown newborns with abnormal intrapartum FHR tracing had elevated umbilical cord plasma EPO (183.5 and 135.2 mIU/ml, respectively; normal = 20.7 mIU/ml) and base deficit, whereas pH, Po2, and 1-minute and 5-minute Apgar scores were significantly lower, compared with appropriately grown newborns with a normal intrapartum course. Among newborns with normal heart rate tracings and FGR, the mean plasma EPO levels were elevated (89.5 mIU/ml), whereas the other parameters were not different from normal. CONCLUSION: Our findings suggest that, although cord blood gases and Apgar scores may reflect subacute and acute events, they are not good predictors of chronic uteroplacental insufficiency. The supplemental use of umbilical cord plasma EPO levels may improve our ability to identify chronic uteroplacental insufficiency. PMID- 10685235 TI - Release of superoxide dismutase activity from human umbilical veins by heparin. AB - OBJECTIVE: This study evaluated superoxide dismutase activity released from human umbilical veins incubated with different doses of heparin and examined at different time points. STUDY DESIGN: Umbilical veins of fresh cords from full term babies were incubated with 175 or 1 U/ml of heparin at one end while the other end was incubated without heparin as control. Specimens were obtained at 10 minutes and 24 hours (high-dose) or at 10 minutes and 60 minutes (low-dose). Superoxide dismutase activity was measured by the cytochrome c method. Results were analyzed using Student's paired t test. RESULTS: A time-dependent release of superoxide dismutase activity into the buffer was observed in both heparin specimens as well as in control specimens. The difference in release in the presence of heparin was of statistical significance, compared with the controls. CONCLUSION: Because heparin is routinely used as an anticoagulant to maintain the patency of umbilical catheters, we conclude that this usage may alter a newborn's response to oxygen free radical damage by changes in superoxide dismutase activity. PMID- 10685236 TI - Preterm infant thermal care: differing thermal environments produced by air versus skin servo-control incubators. AB - OBJECTIVE: Incubator thermal environments produced by skin versus air servo control were compared. STUDY DESIGN: Infant abdominal skin and incubator air temperatures were recorded from 18 infants in skin servo-control and 14 infants in air servo-control (26- to 29-week gestational age, 14 +/- 2 days postnatal age) for 24 hours. Differences in incubator and infant temperature, neutral thermal environment (NTE) maintenance, and infant and incubator circadian rhythm were examined using analysis of variance and scatterplots. RESULTS: Skin servo control resulted in more variable air temperature, yet more stable infant temperature, and more time within the NTE. Circadian rhythm of both infant and incubator temperature differed by control mode and the relationship between incubator and infant temperature rhythms was a function of control mode. CONCLUSION: The differences between incubator control modes extend beyond temperature stability and maintenance of NTE. Circadian rhythm of incubator and infant temperatures is influenced by incubator control. PMID- 10685237 TI - Phototherapy increases hemoglobin degradation and bilirubin production in preterm infants. AB - OBJECTIVE: To compare hemoglobin degradation and bilirubin production before and during phototherapy in preterm infants. BACKGROUND: Hemoglobin is catabolized into globin and heme, which is degraded by microsomal heme oxygenase into equimolar carbon monoxide and biliverdin. Biliverdin is then reduced into bilirubin. CO is excreted exclusively by the lungs; therefore, end-tidal carbon monoxide, corrected for inhaled CO (ETCOc), reflects hemoglobin degradation and total bilirubin production. METHOD: A prospective study design was used, including a study group of 24 preterm infants requiring phototherapy. Infants with hemolytic diseases, sepsis, recent blood transfusion, and infants on mechanical ventilation were excluded. ETCOc was measured in preterm infants before and during phototherapy. Hemoglobin degradation and bilirubin production were calculated by measuring ETCOc. RESULTS: The (mean +/- SD) birth weight of 24 preterm neonates was 1975 +/- 613 gm, gestational age was 32.7 +/- 2.3 weeks, hematocrit was 47.5 +/- 6.2 volume%, and peak bilirubin was 13.1 +/- 3.2 mg/dl. First ETCOc measurements were done at 59.6 +/- 22.2 hours of age immediately before starting phototherapy. The second ETCOc measurements were taken at 13.7 +/ 7.9 hours after starting phototherapy. The second measurement of 2.6 +/- 0.6 ppm (mean +/- SD) was significantly higher than the first ETCOc of 2.1 +/- 0.6 ppm (p < 0.05). CONCLUSION: Phototherapy increases hemoglobin degradation and bilirubin production in preterm infants. PMID- 10685238 TI - Ureaplasma/Mycoplasma-infected amniotic fluid: pregnancy outcome in treated and nontreated patients. AB - OBJECTIVE: To determine if treatment of a positive amniotic fluid culture for mycoplasmal colonization obtained at genetic amniocentesis is associated with improved pregnancy outcome. STUDY DESIGN: A retrospective analysis of 2718 genetic amniocentesis specimens cultured for Ureaplasma/Mycoplasma was undertaken. Specimens were obtained between March 1993 and January 1997. The Irvine culture kit was used to culture all specimens. Data collected included indication for amniocentesis, gestational age at amniocentesis, karyotype, gestational age at delivery, pregnancy outcome, and any antimicrobial treatment. RESULTS: During this time period 44 patients were found to be culture-positive for Ureaplasma/Mycoplasma. Thirty-five were treated with oral erythromycin. Mid trimester loss was 11.4% and 44.4% (p = 0.04) in the treated and untreated groups, respectively. Preterm delivery was similar in the two groups, 19.4% and 20% (p = NS). CONCLUSION: Treatment of an amniotic mycoplasmal colonization with erythromycin was associated with fewer mid-trimester losses after genetic amniocentesis. Preterm delivery rates between the two groups were similar, which may indicate recolonization. PMID- 10685239 TI - The effect of application of aquaphor on skin condition, fluid requirements, and bacterial colonization in very low birth weight infants. AB - OBJECTIVE: To determine the effects of repeated application of an occlusive ointment on the skin of very low birth weight infants. STUDY DESIGN: Nineteen neonates of 26 to 30 weeks gestational age were randomly assigned to receive topical Aquaphor ointment twice daily for 2 weeks or to receive standard skin care. Skin quality, fluid requirements, and skin bacterial colonization counts were assessed. RESULTS: Infants treated with Aquaphor had significantly improved skin condition scores versus controls (p = 0.002). Aquaphor improved skin scores over time (p = 0.012) in treated infants, whereas skin scores of untreated infants worsened before eventually healing. There were no significant differences in total fluid requirements, urine output, serum sodium concentrations, skin bacterial counts, fungal counts, or colonization patterns between treated and control infants in either gestational age cohort. CONCLUSION: Aquaphor ointment, used during the first two postnatal weeks, improved skin condition in infants of 26 to 30 weeks' gestation without changing skin bacterial flora. We speculate that improved skin condition may limit transepidermal water loss and decrease portals of entry for pathogens, thereby potentially decreasing fluid and electrolyte imbalances and sepsis in very low birth weight infants. PMID- 10685240 TI - Predictors of development in premature infants from low-income families: African Americans and Hispanics. AB - OBJECTIVE: The goal of the study was to determine the factors that affect the motor and mental development of premature Latino and African American infants from low socioeconomic backgrounds. STUDY DESIGN: A prospective study of 41 low birth weight (LBW) African American infants along with 82 LBW Hispanic infants examined the factors that influence mental and motor development at 8 months of age. Multiple regression analysis was performed to correlate perinatal, environmental, and demographic variables with mental and motor development using the Bayley scales of infant development. The perinatal variables included birth weight, gestational age, and days of hospitalization. The demographic and environmental variables chosen were: income, education, the home environment, social support, mother-infant interaction, and maternal confidence. RESULTS: The results indicated that, for African American infants, motor development was correlated with the mother's education and the number of days the infant spent in the hospital Mental development for African American infants was predicted by the home environment. For Hispanic infants, the home environment predicted motor scores while the mother-infant interaction was correlated with the mental scores. CONCLUSION: Factors contributing to the development of premature infants vary according to ethnicity, and social variables may be more predictive of development than medical factors. PMID- 10685241 TI - Comparison of DNA probe technology and automated continuous-monitoring blood culture systems in the detection of neonatal bacteremia. AB - OBJECTIVE: The present study assessed the ability of a rapid chemiluminescent DNA probe assay to detect bacterial growth in blood culture specimens before their detection by continuous-monitoring blood culture (CMBC) systems. STUDY DESIGN: Three newborn intensive care units, which are members of the National Institute of Child Health and Human Development Neonatal Research Network, participated in this study. Each center employs an automated CMBC system against which the DNA probe was compared. A total of 1700 blood cultures were analyzed. A 3-ml aliquot of culture medium was removed from each culture during early incubation, processed, and analyzed by the DNA probe. RESULTS: Of 1700 blood cultures, 130 (7.6%) were detected positive by a CMBC system. Coagulase-negative staphylococci (CNS) were present in 90 (69%) of the positive cultures, and 26 (29%) were detected by the DNA probe. Other organisms accounted for the remaining 40 positive cultures, and 31 (78%) of these were detected by the probe assay. Seventy-six percent of all positive cultures were detected by a CMBC system within 24 hours. Ninety-eight percent of all positive cultures were detected by a CMBC system within 48 hours. Of the two organisms that grew in a CMBC system beyond 48 hours, both were CNS and one of these was considered a contaminant. Therefore, 99% of clinically significant organisms were detected by a CMBC system within 48 hours. CONCLUSION: This DNA probe is not sufficiently sensitive to be clinically useful; however, automated CMBC systems are sufficiently sensitive to aid in clinical decision-making regarding the continuance or discontinuance of antibiotic therapy following 48 hours of culture incubation. PMID- 10685242 TI - Sudden deterioration of the newborn infant: II. Diagnosis-based approach in the intensive care unit. PMID- 10685243 TI - Balloon dilatation of the pulmonary valve in a 690-gm neonate with tetralogy of Fallot. PMID- 10685244 TI - Ignac Semmelweis and the etiology of fetal and neonatal sepsis. AB - It is well-known that Ignac Semmelweis discovered the etiology and prophylaxis of puerperal sepsis. However, few historians have focused on his understanding of the pathophysiology of fetal and neonatal sepsis. Based on several key observations, Semmelweis realized that puerperal fever (also known as "childbed fever") could be transmitted to the fetus, especially when the first stage of labor was prolonged and multiple examiners performed vaginal examinations while their fingers were contaminated. This insight was particularly valuable in that it helped him decipher the mystery of puerperal sepsis. This paper presents some of these concepts and supporting evidence. PMID- 10685245 TI - Successful treatment of late-onset infection due to resistant Klebsiella pneumoniae in an extremely low birth weight infant using ciprofloxacin. AB - OBJECTIVE: This paper presents a case in which an extremely low birth weight infant with multidrug-resistant Klebsiella pneumoniae infection was successfully treated with ciprofloxacin and gentamicin. STUDY DESIGN: A clinical case report of a neonate who received broad spectrum antibiotics for possible infection despite negative cultures. The infant developed sepsis and meningitis resulting from multidrug-resistant K. pneumoniae, which was treated with ciprofloxacin and gentamicin. The literature for the use of ciprofloxacin in pediatric patients was reviewed. RESULTS: The infant responded to the antibiotic regimen with sterilization of blood and cerebrospinal fluid; no adverse effects were attributable to the ciprofloxacin. Although ciprofloxacin has been found to cause irreversible injury to cartilage in juvenile laboratory animals, a review of the literature found that this complication occurs rarely if at all in pediatric patients. Ciprofloxacin has been found to be effective in the treatment of multidrug-resistant Gram-negative infections in pediatric patients, including premature infants. CONCLUSION: Ciprofloxacin should be considered in the treatment of neonatal infection caused by multidrug-resistant Gram-negative organisms. Although the published experience with this drug suggests that it is effective and that significant toxicity is not common, its use should be restricted to the treatment of serious infections for which an alternative antibiotics is not available. PMID- 10685246 TI - Haemophilus parainfluenzae sepsis in a very low birth weight premature infant: a case report and review of the literature. AB - Haemophilus parainfluenzae is an unusual cause of invasive bacterial disease and is particularly uncommon as a reported etiology of neonatal sepsis in current large published series. We describe a seriously ill, very low birth weight (VLBW) infant with documented early onset sepsis caused by H. parainfluenzae. We compare our case with those published previously and contrast the clinical presentation of infection in our patient to that of common bacterial pathogens causing neonatal sepsis. Our review suggests many common factors in the pathogenesis of early onset infection by H. parainfluenzae. PMID- 10685247 TI - Testicular torsion in a pre-term neonate. PMID- 10685248 TI - Ventilatory management casebook: recurrent diaphragmatic hernia. PMID- 10685249 TI - Special imaging casebook. Congenital neuroblastoma and cyanotic heart disease. PMID- 10685251 TI - Challenges for neonatology and neonatologists. PMID- 10685250 TI - Necrotizing funisitis with intrapartum umbilical cord rupture. PMID- 10685252 TI - Ultrasonographic differential diagnosis and neurodevelopmental outcome of cerebral white matter lesions in premature infants. AB - OBJECTIVE: To determine the clinical usefulness of recently published ultrasonographic criteria for the differential diagnosis of periventricular hemorrhagic venous infarction (PHVI) versus periventricular leukomalacia (PVL), and its relevance to neurodevelopmental outcome. STUDY DESIGN: From 1992 to 1995, we evaluated 998 very low birth weight infants of which 111 developed cerebral white matter lesions on cranial ultrasonogram examination. An attempt was made to differentiate the lesions into either PHVI or PVL using specific ultrasonographic criteria (Volpe JJ. Brain inury in the premature infant: is it preventable? Pediatr Res 1990; 6:S28-33). Seventy-six patients who survived to discharge constituted the study group. Survivors were followed prospectively with neurologic examinations, visual and auditory screening, and developmental testing. RESULTS: PHVI was diagnosed in 23 patients (30%), PVL in 36 (47%), characteristics of both PHVI and PVL (mixed lesions) in 8 (11%), and persistent periventricular echodensity without cystic change in 9 (12%). Two-year follow-up data were obtained on 57 of 76 (75%) patients. Neurodevelopmental deficits were common in all groups; however, infants with localized PHVI had a mean developmental quotient in the normal range. CONCLUSION: The majority of white matter lesions (77%) can be differentiated as either PHVI or PVL by ultrasonographic criteria, with coexisting features in only 11% of patients. In addition to these lesions, persistent periventricular echodensity was also associated with a high risk of subsequent neurodevelopmental deficit. However, normal development was seen in a subgroup of patients with localized periventricular hemorrhagic venous infarction. PMID- 10685253 TI - Group B streptococcus: to culture or not to culture? AB - OBJECTIVE: To determine if universal Group B Streptococcus (GBS) culturing and antibiotic prophylaxis of obstetric patients decreased the incidence of neonatal early-onset GBS sepsis and mortality and maternal chorioamnionitis. STUDY DESIGN: A time series observational study was conducted to compare the cohort of all obstetric patients delivering at the University of Chicago neonatal center from January 1989 through December 1993, before a GBS surveillance policy existed, with the cohort delivering January 1994 through December 1996, after initiation of a GBS policy. Included in the policy were universal GBS cultures at 28 weeks' gestation, antibiotic prophylaxis at the time of labor for all those with positive cultures and for all with risk factors of preterm delivery, preterm premature rupture of membranes, prolonged rupture of membranes greater than 18 hours, and a previous child affected by GBS or maternal fever in labor. Predictor variables were GBS culturing and antibiotic usage; outcome variables were incidence of GBS sepsis and mortality in the neonates and maternal chorioamnionitis. chi-squared and Fisher exact analyses were used with p < 0.05 being significant. RESULTS: Before the GBS policy, there were 16,272 deliveries with a 2.24/1000 deliveries rate of early-onset GBS sepsis (n = 35); after initiating the GBS policy, 9130 deliveries occurred with an early-onset GBS sepsis rate of 2.29/1000 (n = 20). Early-onset GBS sepsis case fatality rates before and after initiation of the policy were 14.3% and 0%, respectively (p = 0.09). Antibiotic use almost doubled (relative risk = 1.84; confidence interval, 1.74 to 1.93, p < 0.001) over the two time periods, and the relative risk of chorioamnionitis decreased to 0.95 (confidence interval, 0.73 to 0.99, p = 0.04). CONCLUSION: Despite universal GBS culturing and very liberal use of antibiotics in labor, we were unable to effect a statistically significant change in the rate of early-onset GBS sepsis or mortality, and there was only a slightly decreased chorioamnionitis rate. PMID- 10685254 TI - Creamatocrit and the nutrient composition of human milk. AB - OBJECTIVE: We tested the hypothesis that creamatocrit, the length of the cream column separated from milk by centrifugation and expressed as a percentage of the length of the total milk column, is a useful measure of the lipid concentration and the energy content of human milk. STUDY DESIGN: Milk samples from 17 mothers of preterm infants were analyzed prospectively, fresh as well as frozen and thawed, for creamatocrit measurement and nutrient composition. RESULTS: Creamatocrit correlated strongly with lipid concentration and energy content of human milk, fresh or frozen and thawed. The energy content can be calculated from the regression equation: Energy (kcal/dl) = 5.99 x creamatocrit(%) + 32.5 for a fresh sample, and energy (kcal/dl) = 6.20 x creamatocrit(%) + 35.1 for a frozen sample. CONCLUSION: Calculations of energy content from the creamatocrit measurement may be useful for an accurate assessment of energy intake in preterm infants fed human milk. PMID- 10685255 TI - Effect of dexamethasone on lymphocyte subpopulations in premature infants with bronchopulmonary dysplasia. AB - OBJECTIVE: This study was designed to determine the effect of dexamethasone treatment on peripheral blood lymphocyte counts and subpopulations in premature infants with bronchopulmonary dysplasia (BPD). STUDY DESIGN: Peripheral blood lymphocyte subpopulations in 12 premature infants with BPD were analyzed before treatment with a 6-week course of dexamethasone (day 0), on days 3 and 10 of treatment, and 2 weeks after discontinuing dexamethasone therapy (day 56). Lymphocyte immunophenotypes were determined using direct two-color immunofluorescent staining followed by flow cytometry. RESULTS: The percentage of lymphocytes was significantly lower on days 3 (17.55 +/- 2.55) and 10 (20 +/- 11.8) of dexamethasone therapy compared with before (30.36 +/- 6.41) or after treatment. The percentage of T cells was significantly lower on days 3 and 10 of dexamethasone therapy (mean +/- SEM; 58.09 +/- 1.93 and 60.09 +/- 2.47, respectively) compared with before (67.09 +/- 4.24) or after treatment. The absolute number of T cells was significantly lower on day 10 of therapy. The percentage of CD4+ cells was significantly lower on days 3 (38.91 +/- 2.49) and 10 (40.45 +/- 2.24) of therapy, and this decrease persisted after dexamethasone was stopped (36.73 +/- 3.41). The absolute number of CD4 cells was significantly lower on day 10 (1328 +/- 216) of therapy and reached a nadir on day 56 (1143 +/- 106). Similarly, the CD4/CD8 ratio was also significantly lower on days 3 and 10 of treatment (1.56 +/- 0.18 and 1.64 +/- 0.14, respectively) and reached a nadir on day 56 (1.04 +/- 0.13). CONCLUSION: Dexamethasone significantly reduced the percentage and absolute number of lymphocytes, T cells, and CD4 cells, as well as the CD4/CD8 ratio. A reduction in CD4 cells and in the CD4/CD8 ratio persisted 2 weeks after dexamethasone therapy was stopped. In contrast, the absolute number of B cells increased transiently, and CD8 cells were unaffected by dexamethasone. This alteration in lymphocyte subpopulations may help account for the clinically beneficial anti-inflammatory effect of dexamethasone in the treatment of BPD complicated by respiratory failure. The dexamethasone-induced decrease in CD4 cells may also increase the susceptibility of these infants to infection. PMID- 10685256 TI - Low serum cortisol in term newborns with refractory hypotension. AB - OBJECTIVES: The aim of this study is to measure baseline serum cortisol levels and clinical response to glucocorticoid therapy in a group of term infants with refractory hypotension. STUDY DESIGN: Seven term newborns with refractory hypotension were included. Serum cortisol levels were drawn before initiation of glucocorticoid therapy and measured by either fluorescence polarization immunoassay or radioimmunoassay. Baseline blood pressures, heart rate, and inotropes doses were recorded at baseline, then 4, 8, 12, 24, 48, 72, and 96 hours after glucocorticoid therapy. Urine output and volume expanders the infants received were recorded 24 hours before and after glucocorticoid therapy. Dexamethasone was used at a starting dose of 0.2 mg/kg per day divided every 12 hours. The statistical analysis was done using analysis of variance with repeated measures and paired t-test. RESULTS: Serum cortisol levels of the infants ranged from 2.0 to 15.4 micrograms/dl. After initiation of glucocorticoid therapy, there was significant improvement of blood pressure. Vasopressors were rapidly weaned and discontinued within 72 hours. In three of seven infants, no volume expanders were required after initiation of steroids, and none needed volume expanders after 2 days. Urine output increased significantly within 24 hours. All infants survived. CONCLUSIONS: Glucocorticoids improved pressure and stabilized clinical conditions of a group of term newborns with refractory hypotension. Serum cortisol levels of these infants were relatively low. We speculate that a subset of critically ill term infants has relative adrenal insufficiency and glucocorticoid therapy may be essential. PMID- 10685257 TI - Noninvasive validation of tobacco smoke exposure in late pregnancy using end tidal carbon monoxide measurements. AB - OBJECTIVE: To determine whether exposure to tobacco smoke in late pregnancy can be reliably estimated by measuring carbon monoxide (CO) in the mother and newborn breath. STUDY DESIGN: Sixty-eight mothers and their healthy term singleton newborns, delivered at a university-affiliated community hospital in Jerusalem, were enrolled. End-tidal CO (corrected for inhaled air [ETCOc] was measured with a portable automated bedside CO analyzer. ETCOc, cotinine, and carboxyhemoglobin (COHb) levels were compared in 17 smoking, 31 passively exposed, and 20 nonsmoking mothers and their offspring. RESULTS: The mean +/- SD ETCOc was significantly higher in women who smoked than in passively exposed and nonsmoking mothers (8.42 +/- 5.65 vs 1.95 +/- 0.98 vs 1.33 +/- 0.84 ppm. p < 0.0001, respectively). Newborns whose mothers smoked had higher ETCOc levels than those of infants of passively exposed and nonsmoking mothers (10.0 +/- 7.7 vs 2.51 +/- 1.4 vs 1.74 +/- 0.98 ppm, p < 0.0001, respectively). The number of cigarettes smoked by the mother was significantly correlated with maternal ETCOc (r = 0.755, p < 0.00001), and neonatal ETCOc (r = 0.805, p < 0.00001). Maternal ETCOc was highly correlated with neonatal ETCOc (r = 0.857, p < 0.00001), cotinine (r = 0.645, p < 0.00001), and COHb (r = 0.9, p < 0.00001) levels. Birth weight was significantly associated with neonatal ETCOc (p < 0.006) and maternal ETCOc (p < 0.007). CONCLUSION: ETCOc levels in the newborn are well correlated with maternal smoking. Measurements of newborn ETCOc may be used as a noninvasive means to estimate exposure to maternal tobacco smoke immediately before delivery. These measurements will be useful for patient education and research. PMID- 10685258 TI - Contribution of heating, ventilation, and air conditioning airflow and conversation to the ambient sound in a neonatal intensive care unit. AB - Sound reduction strategies in the neonatal intensive care unit (NICU) must focus on the sources of excessive sound. We studied the relative contribution of personnel conversation and heating, ventilation, and air conditioning (HVAC) airflow by measuring several parameters of sound during four environmental conditions: unaltered ambient sound, HVAC airflow off, HVAC airflow and conversation off, and conversation off. All measurements were decreased by interrupting HVAC airflow and conversation. The reduction was greater when conversation was stopped. The method of sound reduction did not change the frequency distribution of sound. Attention to personnel conversation may be effective in lowering sound exposure in the NICU. The contribution of HVAC airflow should be measured in new constructions and renovations. PMID- 10685259 TI - Identifying at risk infants following neonatal extracorporeal membrane oxygenation. AB - OBJECTIVE: To identify infants at risk of death and abnormal neurodevelopmental outcome following extracorporeal membrane oxygenation (ECMO) in the neonatal period. METHODS: The medical records of 82 neonates treated with ECMO were reviewed to evaluate risk of death. All survivors were followed by neurologic examinations and tested using the Bayley Scales of Infant Development or McCarthy Scale of Children's Abilities, and risk for abnormal neurodevelopmental outcome was assessed. RESULTS: The overall survival was 91% (75 of 82). The mean gestational age and birth weight of nonsurvivors were lower than those of survivors (37 +/- 1 weeks vs 40 +/- 0 weeks; 2734 +/- 230 vs 3325 +/- 69 gm, p < 0.05). Infants who were lost to follow-up (16%) did not differ from those with follow-up in demographic variables or clinical indicators of illness severity. Thirty-five of 63 infants (56%) with follow-up had normal neurodevelopmental outcome. Risk of abnormal outcome was higher in infants requiring assisted ventilation for > or = 15 days (relative risk [RR] 5.5; 95% confidence interval [CI] 2.0 to 14.8), supplemental oxygenation for > or = 22 days (RR 3.1; 95% CI 1.3 to 7.6), and black race (RR 8.9; 95% CI 1.3 to 62.9). None of the neuroimaging studies accurately predicted the neurodevelopmental outcome of these infants. CONCLUSION: We conclude that ECMO in critically ill infants is associated with good survival. The need for prolonged respiratory support may help in identifying infants at risk for abnormal neurodevelopmental outcome. PMID- 10685260 TI - Impact of pediatric ethics consultations on patients, families, social workers, and physicians. AB - OBJECTIVE: To evaluate whether health providers and families find ethics consultations helpful in identifying, analyzing, and resolving ethical problems. STUDY DESIGN: Forty consecutive pediatric ethics consultations at the San Diego Children's Hospital and Health Center were evaluated retrospectively through chart reviews and structured interviews with physicians, social workers, and family members. RESULTS: In 23 of 40 cases, physicians or social workers were successfully interviewed. Of these 23 cases, four family interviews were completed. Over 90% of physicians and social workers found the ethics consultation to be helpful and would recommend an ethics consultation to others in the same circumstances. Two of the four families were strongly dissatisfied with the consultation and identified miscommunication of the ethics consultant's role as a major problem. CONCLUSION: The disparity observed in this study between satisfaction levels of health providers and families raises concerns. More studies that evaluate ethics consultations are needed, especially those that are designed prospectively and explore both these perspectives. PMID- 10685261 TI - Maternal hypoglycemia: is it associated with adverse perinatal outcome? AB - OBJECTIVE: To determine if maternal hypoglycemia is associated with adverse perinatal outcome, particularly low birth weight. STUDY DESIGN: In this prospective study, all patients after 24 weeks' gestation were screened for gestational diabetes using 50 gm of glucola (oral) followed by a 1-hour plasma glucose measurement and hypoglycemia was defined as < or = 88 mg/dl. RESULTS: In these 426 women the mean (+/- SD) 1-hour plasma glucose value was 99.8 +/- 22.7 mg/dl. Of these, 16 were diagnosed with gestational diabetes and 46 were lost to follow-up leaving 364 patients; 116 with hypoglycemia and 248 with euglycemia. Women with hypoglycemia weighed less at the beginning of pregnancy and at delivery, but total weight gain during pregnancy was similar between both groups. There was no difference between groups in maternal symptomatology, birth weight, or the rate of fetal growth restriction. CONCLUSION: Hypoglycemia on the 1-hour glucola screen is not predictive of fetal growth restriction or other adverse perinatal consequence. PMID- 10685262 TI - Unilateral do-not-resuscitate order in the neonatal intensive care unit. PMID- 10685263 TI - Treatment of neonatal infection caused by coxsackievirus B3. AB - Four male infants with early neonatal infection caused by coxsackievirus B3 (presumed in one case) exhibited severe thrombocytopenia and liver dysfunction at presentation. The three infants who were administered human normal immunoglobulin within 3 days of disease onset survived, while the fourth infant, who received the preparation 6 days after disease onset, died. PMID- 10685264 TI - Curling's ulcer in association with staphylococcal scalded skin syndrome in the neonate. PMID- 10685265 TI - Placental pathology casebook. Chorangiosis of the placenta increases the probability of perinatal mortality. AB - Two apparent acute problems that may occur in labor, nuchal cord and placental abruption, were associated with chorangiosis of the placenta. The importance of complete placental examination in perinatal mortality is re-emphasized. The association of apparent acute obstetrical conditions, e.g., nuchal cord and placental abruption with chorangiosis of the placenta, may be the cause of fetal newborn deaths that were previously assumed to be issues of labor management. PMID- 10685267 TI - Informed consent: are we there? PMID- 10685266 TI - Perinatal/neonatal transport casebook. A 3-week-old female infant with a cough and limp spells. PMID- 10685268 TI - Neurodevelopmental outcome of prematurely born children treated with recombinant human erythropoietin in infancy. AB - OBJECTIVE: To compare the neurodevelopmental outcome of premature infants treated with recombinant human erythropoietin with that of control infants. STUDY DESIGN: A total of 20 treated infants and 20 control infants who had completed randomized, double-blind, placebo-controlled studies of recombinant human erythropoietin as treatment for anemia of prematurity were followed for growth and developmental outcome in an intensive care nursery follow-up program. Infants were assessed by standard developmental tests. RESULTS: No differences were found between groups for neurologic outcome, cognitive outcome, or growth patterns. All infants treated with recombinant human erythropoietin were neurologically normal. The rate of cognitive deficits was similar in the two groups. CONCLUSION: In this small sample we did not see differences in neurodevelopmental outcome between infants treated with recombinant human erythropoietin and control infants. PMID- 10685269 TI - Relationship of amniotic fluid markers of intra-amniotic infection with histopathology in cases of preterm labor with intact membranes. AB - OBJECTIVE: To evaluate the correlation of amniotic fluid (AF) markers (AFMs) of intra-amniotic infection with histopathologic findings in cases of preterm labor with intact membranes, between 22 and 36 weeks' gestation. STUDY DESIGN: We reviewed the charts of patients admitted in preterm labor with intact membranes between January 1993 and December 1996. Those having amniocentesis were identified, and AFMs were compared with histopathology in patients who delivered within 48 hours of the amniocentesis. The AFMs evaluated were glucose, polymorphonuclear leukocytes, Gram stain, and culture. All placentae were reviewed by a single pathologist blinded to the AF findings. Histologic evidence of acute inflammation was defined by findings of both subchorial intervillositis and marginating choriodeciduitis. The sensitivities, specificities, and positive and negative predictive values of the various AFMs were calculated. RESULTS: Of 556 women with intact membranes presenting in preterm labor, 181 (32.6%) had amniocentesis and 88 delivered within 48 hours of the amniocentesis. Histopathologic chorioamnionitis was seen in 53 patients (60.2%). The findings (with their sensitivity, specificity, and positive and negative predictive values) were: polymorphonuclear leukocytes at > 10/high-power field (22.6%, 97.2%, 92.3%, and 46.1%), positive Gram stain (26.4%, 94.6%, 87.5%, and 47.3%), culture (28.3%, 92.1%, 83.3%, and 47.9%), and glucose of < 15 mg/dl (28.3%, 94.6%, 88.2%, and 47.9%), respectively. Using a receiver-operator characteristic curve for different level of AF glucose, a glucose level of < 20 mg/dl was the most sensitive AF predictor of histologic chorioamnionitis. CONCLUSION: Histopathologic evidence of chorioamnionitis was present in 60.2% of cases of preterm births due to preterm labor in women who at our institution were offered and accepted amniocentesis and subsequently delivered within 48 hours. AFMs may be useful predictors of histologic chorioamnionitis. The most efficient AFM for chorioamnionitis in this group of patients was glucose at < 20 mg/dl. PMID- 10685270 TI - A randomized trial of nasopharyngeal-synchronized intermittent mandatory ventilation versus nasopharyngeal continuous positive airway pressure in very low birth weight infants after extubation. AB - OBJECTIVE: To prospectively compare the incidence of respiratory failure in premature infants randomized to receive either nasopharyngeal continuous positive airway pressure (NPCPAP) or nasopharyngeal-synchronized intermittent mandatory ventilation (NP-SIMV) in the immediate postextubation period. STUDY DESIGN: This is a prospective study of very low birth weight (VLBW) infants randomized at the time of extubation to receive either NPCPAP or NP-SIMV in a university-based level III neonatal intensive care unit. Statistical analysis were performed with the Mann-Whitney U test for continuous and ordinal variables, and with the chi squared test or Fisher's exact test for categorical variables. RESULTS: A total of 41 VLBW infants were studied; 19 were in the NPCPAP group, and 22 were in the NP-SIMV group. Respiratory failure after extubation in the NP-SIMV group was significantly lower that in the NPCPAP group (5% vs 37%, respectively (p = 0.016). No statistically significant differences between groups with regard to demographics, severity of initial illness and associated complications, time to extubation, ventilatory management before extubation, weight, age, or nutritional status at the time of extubation were noted. PMID- 10685271 TI - Effects of antenatal corticosteroids on blood pressure in very low birth weight infants during the first 24 hours of life. AB - OBJECTIVE: We tested the hypothesis that prenatal glucocorticoids significantly increase mean arterial blood pressure in very low birth weight preterm infants during the first 24 hours after birth. STUDY DESIGN: Prospectively collected data from 178 inborn infants with birth weights between 500 and 1499 gm were examined. A total of 80 infants were born to mothers treated with corticosteroids (birth weight: 1057 +/- 271 gm, gestational age: 28.0 +/- 2.6 weeks), and 98 infants were untreated controls (birth weight: 1030 +/- 280 gm, gestational age: 28.0 +/- 2.8 weeks). The study setting was a university-based tertiary care center for newborn intensive care. RESULTS: Mean blood pressures on admission and at 3, 6, 12, 18, and 24 hours were significantly higher in steroid-treated infants. Steroid-treated infants received significantly less volume expansion (3.8 +/- 8.5 ml/kg versus 14.4 +/- 20.7 ml/kg; p < 0.001) than controls. Vasopressor support was also reduced in the steroid group (2.5% versus 11.5%; p < 0.05). CONCLUSION: Antenatal steroids are associated with both a higher mean systemic blood pressure and a decreased use of vasopressors and plasma expanders in very low birth weight infants during the first 24 hours after birth. This effect is not limited to infants of < 1000 gm; it is also significant in infants with a birth weight between 1000 and 1499 gm, and is already detectable in the first hours of life. We speculate that this finding may contribute to the mechanism of steroid protection against conditions such as intraventricular hemorrhage. PMID- 10685272 TI - Skin care management practices for premature infants. AB - OBJECTIVE: To describe current skin care practices for preterm infants in neonatal intensive care units in the United States. We hypothesized that there would be little consensus among facilities. STUDY DESIGN: Neonatal intensive care units (n = 823) listed in the 1996 United States Neonatologists Directory (American Academy of Pediatrics, Section on Perinatal Pediatrics) were sent a 28 question survey dealing with many aspects of neonatal skin care along with descriptive data about their neonatal intensive care unit. Descriptive data analysis was performed. RESULTS: A total of 305 surveys were returned (37% return rate); of these, 241 of the respondents reported admitting infants weighing < or = 1000 gm. Some neonatal skin care practices showed wide consensus (> 70%) (e.g., scrub procedure for staff; use of a skin barrier under tapes/adhesives), whereas other practices showed little consensus (< 30%) (e.g., routine surveillance cultures; use of Aquaphor). CONCLUSION: Consensus on skin care practices was not found among neonatal intensive care units. Data from this survey can be used to develop studies to examine whether certain skin care management practices can improve neonatal outcomes. PMID- 10685273 TI - Effect of the maternal care manual of the perinatal education programme on the ability of midwives to interpret antenatal cards and partograms. AB - OBJECTIVE: To assess the ability of midwives to interpret antenatal cards and partograms correctly following completion of the Maternal Care Manual of the Perinatal Education Programme. STUDY DESIGN: We conducted a prospective, controlled trial in a study town and two control towns in the Eastern Cape Province of South Africa. All 93 midwives caring for pregnant women in the three towns were included in the study. Samples were compared using the two-tailed Student's t-test. RESULTS: The marks achieved by the study group for questions from the antenatal card and the partogram improved by 33.0% (p < 0.001) and 17.5% (p = 0.001), respectively. No changes were observed in the control group. CONCLUSION: Midwives that studied the Maternal Care Manual significantly improved their ability to interpret clinical information and apply knowledge. If this ability is applied in clinical practice, a reduction in maternal and perinatal deaths is possible. PMID- 10685274 TI - Survival, intracranial lesions, and neurodevelopmental outcome in infants with congenital diaphragmatic hernia treated with extracorporeal membrane oxygenation. AB - OBJECTIVE: Before the use of extracorporeal membrane oxygenation (ECMO), infants with a severe form of congenital diaphragmatic hernia (CDH) had a high mortality and morbidity. Recent studies have shown an improvement in the survival of these infants after ECMO treatment; however, the existing data do not provide sufficient informations regarding the quality of survival and developmental outcome of these infants. The objective of this study was to evaluate survival, intracranial lesions, and the neurodevelopmental outcome of infants with CDH treated with ECMO. METHODS: We retrieved data for 51 (n = 51) infants with CDH who were treated with ECMO at Huntington Memorial Hospital between 1985 and 1994. Their mean gestational age was 38.5 +/- 2.4 weeks (mean +/- SD); their mean birth weight was 3170 +/- 620 gm. Vital signs, arterial blood gases, chest radiographs, cranial and cardiac ultrasonography were routinely obtained before ECMO treatment. Cranial ultrasounds were performed daily on all infants while on ECMO; computerized tomography scans were obtained on all infants after completion of ECMO treatment. The surviving infants were followed at our neonatal follow-up clinic for neurodevelopmental assessment. RESULTS: A total of 39 infants were placed on venoarterial ECMO and 12 infants were placed on venovenous ECMO; a total of 35 infants had CDH repair before ECMO, whereas 16 infants had delayed surgery. A total of 31 infants (61%) survived. The infants who survived had a mean pH of 7.33 +/- 0.20, mean airway pressure of 19.6 +/- 5.8 cm H2O, and an oxygenation index (OI) of 87 +/- 55 before ECMO intervention. The infants who expired (n = 20) had a mean pH of 7.31 +/- 0.15, mean airway pressure of 23.1 +/- 5.5 cm H2O, and a mean oxygenation index of 127 +/- 56 before ECMO treatment. Before ECMO, survivors had a significantly lower oxygenation index and a higher Pao2 compared with nonsurvivors (p < 0.01). A total of 18 infants (35%) had abnormal central nervous system findings. Of the 51 infants, 10 had ventricular dilatation, 6 had intracranial hemorrhage, and 11 had focal or diffuse cerebral atrophy diagnosed by computerized tomography scan or at autopsy (1 patient had an infarct). Eight infants had more than one central nervous system abnormality. A total of 16 survivors had a neurodevelopmental evaluation at 12 months, and 11 of these survivors were evaluated at 24 months of age (Bayley Scales of Infant Development). The developmental progress of these infants falls within the low average range of cognitive and motor abilities. Their mean Bayley Mental Developmental Index was 85 +/- 25 (50 to 145) at 24 months; their Psychomotor Developmental Index was 89 +/- 21 (50 to 113) at 24 months of age. Follow-up at 4 and 6 years of age is in progress. CONCLUSION: Our preliminary findings indicate that 35% of infants with severe CDH requiring ECMO had central nervous system abnormalities (intracranial lesions, including ventricular dilatation). The survival rate in our study population is consistent with recent reports. As a group, infants with severe CDH display mild neuromotor and cognitive delay in development at 24 months of age. PMID- 10685275 TI - Alveolar capillary dysplasia: diagnostic potential for cardiac catheterization. AB - OBJECTIVE: Alveolar capillary dysplasia is a rare cause of persistent pulmonary hypertension of the newborn. Infants with this condition die despite maximal medical intervention including inhaled nitric oxide therapy and extracorporeal membrane oxygenation. To date, diagnosis of this lethal condition was made by open lung biopsy or during postmortem examination. We examined the possibility that distinct cardiac catheterization findings could be used in the diagnosis of this lethal disorder. STUDY DESIGN: We present three infants with fatal persistent pulmonary hypertension of the newborn refractory to extracorporeal membrane oxygenation and inhaled nitric oxide therapy, two with postmortem autopsy confirmation of alveolar capillary dysplasia. Each infant underwent cardiac catheterization to complete the diagnostic evaluations. RESULTS: Significant right ventricular hypertension and normal pulmonary venous return were demonstrated, but a markedly diminished or absent capillary blush phase was noted in each infant. This finding is distinct from the normal capillary blush seen in infants with persistent pulmonary hypertension of the newborn of other etiologies. CONCLUSION: Cardiac catheterization may provide a useful alternative to tissue examination in the diagnosis of alveolar capillary dysplasia. PMID- 10685276 TI - Focal neurological manifestations following aberrant central venous catheter placement. AB - An infant developed focal tonic clonic movements of both lower limbs while receiving total parenteral nutrition through a left saphenous percutaneous central venous catheter. Radiographic studies using a contrast confirmed that the catheter tip was located in the ascending lumbar vein in close proximity to the epidural space. Withdrawal of the catheter abated all clinical symptoms. This case emphasizes the need to confirm central venous catheter placement and illustrates yet another risk associated with the infusion of parenteral alimentation. PMID- 10685277 TI - Dissemination of the kangaroo method in Germany. PMID- 10685278 TI - Gestation versus outcome table for parents of extremely premature infants. AB - Parents of extremely premature babies may have problems understanding and remembering information on likely outcome. We have constructed a simplified evidence-based "gestation versus outcome" table that could be offered to the parents. Such a table may promote consistency in the information given to parents by different members of the perinatal team. PMID- 10685279 TI - Lethal pulmonary hypoplasia and hydrocolpos with transverse vaginal septum in a newborn: a case report and review of the literature. AB - This is a case report of an unusual cause of Potter sequence. Autopsy showed lethal pulmonary hypoplasia in association with a transverse vaginal septum, hydrocolpos, and a secondary obstructive uropathy. PMID- 10685280 TI - Association of maternal sertraline (Zoloft) therapy and transient neonatal nystagmus. PMID- 10685281 TI - Bacteremia, meningitis, and brain abscesses in a hospitalized infant: complications of Pseudomonas aeruginosa conjunctivitis. AB - This report describes a preterm infant hospitalized in a neonatal intensive care unit who developed Pseudomonas aeruginosa conjunctivitis associated with bacteremia, meningitis, and multiple brain abscesses. P. aeruginosa conjunctivitis can rapidly progress to an invasive eye infection, such as corneal ulceration or endophthalmitis, leading to poor vision or blindness. Progression of this infection may lead to systemic disease. However, as illustrated in this report, P. aeruginosa conjunctivitis may be associated with the development of systemic complications such as bacteremia and meningitis in the absence of invasive eye disease. P. aeruginosa is a relatively common cause of conjunctivitis in hospitalized preterm and low birth weight infants. Given the severity of the ocular and systemic complications of Pseudomonas conjunctivitis, clinicians are reminded that prompt detection and treatment of neonatal conjunctivitis is critical. PMID- 10685282 TI - Umbilical cord blood gases casebook. Interpreting umbilical cord blood gases, V. PMID- 10685283 TI - Special imaging casebook. Congenital adrenal neuroblastoma with renovascular hypertension. PMID- 10685284 TI - Radiology casebook. Intussusception in a premature infant: a case report. PMID- 10685285 TI - Is community important for health?: examining the biopsychosocial interface. AB - The United States has created the most expensive, technologically advanced medical system in the world. Health outcomes, however, fail to achieve results commensurate with investment. After identifying the scope of population health concerns untreated by the current U.S. healthcare system, an explanatory model suggests that the relational basis of health and adaptation has been neglected by providers and policymakers. Finding root sources of health in the strength of relationships between individuals and within communities, recommendations are made for applying an integrated model of personal, community, and national health. PMID- 10685286 TI - How to help parents of young children: the touchpoints model. PMID- 10685287 TI - Community efforts to improve perinatal care: a clinical pathologic conference. PMID- 10685288 TI - National Perinatal Association (NPA) debate: should the legislature regulate perinatal practice? PMID- 10685289 TI - Two tools for well-being: health systems and communities. AB - Medical systems are tools of limited power to improve health status in modern societies. The associational tools of local communities are now the most important health-giving resources. Health professionals must learn the nature of these tools and understand their relation to and support of them. PMID- 10685290 TI - From public health to population health: epidemiological yardsticks for perinatal care. AB - Perinatal epidemiology and health services research have focused on specific health outcomes or disease states ("key" indicators and/or sentinel events), clients of public programs, and clinical service populations. In this paper, the concept of "population health" is defined and operationalized for the broad perinatal healthcare context within the framework of population health informatics. Traditional indicators and outcome measures for perinatal health are compared with some newer measures and assessed for their relevance to population health. A hypothetical model for the implementation of population health informatics for maternal and child health is described; in addition, and some obstacles to the practice of population health are identified, with strategies to overcome these obstacles. PMID- 10685291 TI - The relational basis of health: woman's voice. AB - Using voice as a metaphor, this paper will speak to the necessity of health care providers to "hear" the language of women, promote validity to their experiences, and facilitate their relational connections in their evolving roles as mothers. PMID- 10685292 TI - Efficacy and justice: the importance of medical research and tertiary care to social disparities in infant mortality. PMID- 10685293 TI - Communication gap. PMID- 10685294 TI - Influence of postdatism and meconium on fetal erythropoietin. AB - OBJECTIVE: To determine whether fetal erythropoietin (Epo) concentrations are increased in pregnancies extending beyond 41 weeks' gestation and whether this is influenced by the presence of meconium-stained amniotic fluid. METHODS: Epo concentrations were measured in 116 fetal umbilical cord blood samples from otherwise uncomplicated pregnancies between 37 to 43 weeks' gestation during the period of October 1996 to October 1997. An enzyme-linked immunosorbent assay kit was used to measure Epo. Maternal demographics and birth outcomes including Apgar score, cord blood pH, and base deficit were obtained. Fetuses born between 41 and 43 weeks' gestation (post-term) were compared with matched controls born between 37 and 40 weeks' gestation (term). In addition, both post-term and term fetuses with meconium-stained amniotic fluid were compared with matched controls without meconium. RESULTS: Post-term fetuses without meconium had significantly higher Epo levels compared with term fetuses (mean +/- SEM: 50.6 +/- 6.5 versus 29.5 +/- 3.3 mIU/ml, p = 0.002). When matched for gestational age, fetuses with meconium stained amniotic fluid had significantly greater Epo concentrations compared with controls without meconium (post-term, 80.7 versus 50.6 mIU/ml; term, 61.4 versus 29.5 mIU/ml; p < 0.05). However, no significant difference in Epo levels was found between post-term fetuses with meconium and term fetuses with meconium (80.7 +/- 15.7 mIU/ml versus 61.4 +/- 12.8 mIU/ml, respectively). Mean cord blood pH and base deficit values for all groups were within normal clinical range. CONCLUSION: Cord blood Epo concentrations were significantly increased in pregnancies extending beyond 41 weeks. Irrespective of gestational age, meconium stained amniotic fluid was associated with a significant rise in Epo. High Epo levels in these pregnancies imply subacute or chronic fetal hypoxia. Close clinical monitoring of post-term fetuses and those with meconium-stained amniotic fluid is warranted. PMID- 10685295 TI - Human immunodeficiency virus (HIV)-related risk-taking behaviors in women attending inner-city prenatal clinics in the mid-west. AB - OBJECTIVE: Federal guidelines recommend the provision of human immunodeficiency virus education to all attendees of prenatal clinics. The current study was conducted to assess risk-taking behaviors among urban women voluntarily pursuing prenatal care. DESIGN AND SETTING: African American women attending urban prenatal clinics in Cleveland, Ohio were subjected to an extensive interview before receiving an experimental AIDS education curriculum. The interview sought detailed information regarding demographics, lifetime and recent sexual activity, condom use, and lifetime and recent illicit drug use. RESULTS: A total of 1017 women were interviewed; of those women, approximately 73% were single. The majority had a monthly income of less than $500. A total of 66% had only one partner in the past year, and almost 90% had < or = 1 partner in the past 6 months. A total of 98% identified a main partner. Nearly all subjects were at least fairly certain that this partner did not use intravenous drugs, and 71% were at least fairly certain that he was monogamous. Only 19% used condoms most or all the time. Intravenous drug use among study subjects was very infrequent. CONCLUSION: These data indicate that inner-city Cleveland women seeking prenatal care are largely monogamous around the time of their gestation, and that a history of intravenous drug use is infrequent. They suggest that prenatal counseling in urban clinics will need to address women who largely are engaged in single-partner relationships at the time of the intervention. PMID- 10685296 TI - Predictors of symptomatic urinary tract infection after 20 weeks' gestation. AB - OBJECTIVE: To identify predictors of symptomatic urinary tract infection (UTI) after 20 weeks' gestation. STUDY DESIGN: A retrospective cohort analysis was conducted of all deliveries at three North Carolina hospitals between 1990 and 1993. A total of 7403 deliveries remained after exclusions (pre-pregnancy diabetes, HIV-positive, structural urologic abnormalities, no prenatal care) and restrictions (black or white race, county of residence). Cystitis and pyelonephritis were identified by clinician diagnosis. Multiple logistic regression was conducted. RESULTS: Prior UTIs (both before and earlier in pregnancy), nonprivate clinics, and a history of chlamydia (white women only) doubled the risk of symptomatic UTIs after 20 weeks' gestation. The strongest predictor of pyelonephritis was prior antenatal UTIs (adjusted incidence odds ratio = 5.3, 95% confidence interval of 2.6-11.0), followed by less education (< 12 years), a history of chlamydia, nonprivate clinics, illicit drug use, sickle cell hemoglobinopathy, and being unmarried. CONCLUSION: Medical history and demographic factors predict cystitis and pyelonephritis after 20 weeks' gestation. Prospective studies of pyelonephritis predictors and screening strategies are warranted. PMID- 10685297 TI - Sound transmission into incubators in the neonatal intensive care unit. AB - OBJECTIVE: To measure the attenuation of sound by modern incubators. STUDY DESIGN: LEQ, LMAX, LPEAK, and frequency distribution were measured simultaneously inside and outside two recent model incubators. RESULTS: The attenuation of sound (outside minus inside) was 15 to 18 dBA with the motor off and 4 to 8 dBA with the motor on. There was a significant difference between incubators in their attenuation of sound. Octave band analysis showed attenuation in frequency bands of > 31.5 Hz with the motor off. With the motor on, the sound level inside the incubator was higher than outside at frequency bands of < 250 Hz. CONCLUSION: Caring for infants inside modern incubators reduces "averaged" sound exposure to levels near those recommended for the neonatal intensive care unit. Lower frequency sounds are louder inside the incubator and arise from the incubator motor. PMID- 10685298 TI - Nephrocalcinosis in premature infants: variability in ultrasound detection. AB - OBJECTIVE: To measure variability among radiologists in the ultrasound diagnosis of nephrocalcinosis in premature infants. METHODOLOGY: In this prospective multicenter study, renal ultrasounds were performed on 54 very low birth weight infants using a 5.0- and 7.5-MHz transducer, and these ultrasounds were read independently by three radiologists. kappa coefficients were calculated to assess variability in identification of nephrocalcinosis among the radiologists. RESULTS: The kappa coefficient (+/- confidence intervals) using a 5.0-MHz transducer was 0.143 (0.108, 0.178); using the 7.5-MHz transducer, the kappa coefficient was 0.268 (0.243, 0.293). All three radiologists agreed in their identification of nephrocalcinosis on 3 of 54 ultrasounds using a 5.0-MHz transducer; a total of 6 of 54 ultrasounds obtained using a 7.5-MHz transducer were read as positive by all three radiologists. CONCLUSION: There is significant variability among radiologists in the ultrasound identification of nephrocalcinosis in premature infants; a 7.5-MHz ultrasound transducer is associated with less variability in recognizing this lesion. PMID- 10685299 TI - Relationship between perinatal counseling and incidence of breastfeeding in an inner-city population. AB - OBJECTIVE: We hypothesized that the cost of a lactation program can be reduced without significantly affecting the incidence of breastfeeding. STUDY DESIGN: We conducted a retrospective analysis of breastfeeding among all 7942 mothers whose neonates were admitted to the well baby nursery at Jacobi Medical Center (JMC) over a 44-month period. We used multiway frequency analysis to compare the incidence of breastfeeding in three successive models of counseling: (1) full time lactation coordinator, (2) obstetric personnel trained in breastfeeding counseling and full-time lactation coordinator, and (3) obstetric personnel and half-time lactation coordinator. Mothers were further classified into three groups according to location of prenatal care and attendance at breastfeeding education sessions. RESULTS: Breastfeeding increased with the initiation of education and the involvement of obstetric personnel and did not significantly decrease when the lactation coordinator became half-time. The transition to model 3 resulted in decreased costs without significantly affecting the incidence of breastfeeding. Breastfeeding was significantly associated with counseling by obstetric personnel, with prenatal care at JMC, and with breastfeeding education sessions. CONCLUSION: Involving obstetric personnel in breastfeeding counseling may enhance the effectiveness of a lactation program. In our population, the most cost-conscious model included counseling by trained obstetric personnel and a half-time lactation coordinator. PMID- 10685300 TI - Weekly telephone contact does not enhance the compliance of home apnea monitoring. AB - OBJECTIVE: To evaluate the effect of weekly telephone contact with families in enhancing the use of home apnea monitors. STUDY DESIGN: This was a prospective, randomized, single-blinded study of 65 infants who were prescribed home apnea monitoring at the time of initial discharge from the hospital. Exclusion criteria included participation in any other study involving home monitoring or nonavailability of home telephone. Infants were randomized either to the "standard" or "telephone" group by a stratified balanced block technique. All families were instructed to use the monitor during the first 4-week period at all times except during bathing and during the second 4-week period at all unattended times and at night. The families in the telephone group were contacted weekly for 8 weeks. The telephone interview reviewed the events of the previous week but did not include specific encouragement to use the monitor. Both groups received routine pediatric care and follow-up at our high-risk premature clinic. The primary outcome measure was compliance measured as the percentage of time as well as the hours per day that the infant spent on the monitor as recorded by the documented monitor. RESULTS: The telephone (n = 30) and standard (n = 32) groups were similar (p > 0.10) with respect to birth weight (1567 +/- 778 versus 1710 +/ 777 gm), gestational age (30.9 +/- 4.2 versus 31.1 +/- 4.6 weeks), maternal age (24.9 +/- 6.0 versus 25.3 +/- 5.4 years), and commercial insurance (46.7% versus 46.9%), a marker of higher socioeconomic status. Compliance of the telephone versus the standard group was similar during the first 4-week period (74.7 +/- 24.9 versus 75 +/- 27.8%, p = 0.85) (17.9 +/- 5.9 versus 18.2 +/- 6.6 hours/day), the second 4 week period (63.4 +/- 29.1 versus 58.9 +/- 30.9%, p = 0.59) (15.2 +/ 7.0 versus 14.1 +/- 7.4 hours/day) and the entire 8-week period (69.3 +/- 24.7 versus 67.7 +/- 26.2%, p = 0.82, Mann-Whitney U-test) (16.7 +/- 6.0 versus 16.1 +/- 6.5 hours/day), respectively. An abnormal pneumocardiogram at the time of discharge was the only identified factor that improved the compliance for the entire 8-week period (73.1 +/- 22 versus 52.1 +/- 28.5%, p = 0.02) (17.5 +/- 5.2 versus 12.5 +/- 6.8 hours/day) and the first 4-week period of monitoring (81.7 +/ 22.9 versus 59.5 +/- 31.3%, p = 0.01) (19.6 +/- 5.5 versus 14.2 +/- 7.5 hours/day). CONCLUSION: Weekly telephone contact, without specific encouragement to use the monitor, did not improve compliance. Compliance was greater in subjects who had abnormal pneumocardiogram results at the time of discharge from hospital regardless of their telephone/standard group assignment. We speculate that in this already compliant population, more targeted advice is necessary to increase compliance. PMID- 10685301 TI - Use of positioning to reduce the severity of neonatal narcotic withdrawal syndrome. AB - OBJECTIVE: This study tested the hypothesis that highly fretful, narcotic withdrawing neonates experience less distress in a prone-lying position than comparable, supine-lying neonates. STUDY DESIGN: Equivalent numbers of randomly assigned, narcotic-withdrawing newborns were assigned to prone-lying (n = 25) or supine-lying (n = 23) conditions. Subjects in the two groups were similar with regard to gestational age, birth weight, and clinical presentation. Peak and mean withdrawal severity, as measured by Neonatal Abstinence Scoring System (NASS) scores and daily caloric intake, were compared between supine and prone groups by Wilcoxon's two-sample test. RESULTS: The prone-lying neonates had lower peak NASS scores (p < 0.0001), lower mean NASS scores (p < 0.0001), and lower caloric intake (p < 0.001) than supine-lying, narcotic-withdrawing newborns. CONCLUSION: The fretfulness associated with neonatal withdrawal and other stressful conditions can be moderated by laying the affected infant prone. The pronate quieting response is a significant, endogenous source of neonatal pacification. PMID- 10685302 TI - Maternal and fetal factors related to abnormal amniotic fluid. AB - OBJECTIVE: The objective of this study was to identify maternal and infant characteristics related to alteration of amniotic fluid volume at birth. STUDY DESIGN: A series of 27,145 consecutive malformed newborn infants from the Spanish Collaborative Study of Congenital Malformations (ECEMC) was analyzed. From this total, 3.01% were found to have oligohydramnios and 3.69% were found to have polyhydramnios. RESULTS: As expected, renal/urinary tract and lung defects were associated with oligohydramnios, whereas esophageal and intestinal atresias, neural tube defects, and other central nervous system malformations were associated with polyhydramnios. In addition, other defects such as cardiovascular anomalies, hydrocephaly, and microcephaly were also related to abnormalities of amniotic fluid volume. After excluding the defects whose association to oligo- or polyhydramnios is well recognized, we compared the frequency of different variables among them and with infants with a normal volume of amniotic fluid. In comparison with infants with normal amniotic fluid volume, the groups with oligo- and polyhydramnios had lower birth weight, shorter gestational age and umbilical cord, higher parental ages, and a greater frequency of spontaneous abortions. The differences were more marked for weight in newborn infants with oligohydramnios, and for gestational age, umbilical cord length, number of previous pregnancies, and spontaneous abortions in polyhydramnios cases. Placental weight was lower in oligohydramnios cases than in infants with normal amniotic fluid, and higher in polyhydramnios cases. Parental consanguinity and twinning were more frequent in polyhydramnios. Maternal morbidity was higher in both groups with abnormal amniotic fluid volume, especially for acute diseases such as hypertension, diabetes mellitus, and gestational diabetes. Chromosomal aberrations were more frequent in the oligo- and polyhydramnios groups than in cases with a normal volume of amniotic fluid, which supports the suggestion of performing prenatal cytogenetic analysis in any pregnancy complicated by an abnormal volume of amniotic fluid. CONCLUSION: The fact that all of these results are similar in the control group of healthy infants suggests that at least some of the variables associated with abnormal amniotic volume could be considered as causal factors altering the production of fluid. PMID- 10685303 TI - The fetus with gastroschisis managed by a trial of labor: antepartum and intrapartum complications. AB - OBJECTIVE: To assess the rate of antepartum and intrapartum complications of fetuses with antenatally diagnosed gastroschisis managed in a center that advocates a trial of labor. STUDY DESIGN: A retrospective review. The medical records of 49 fetuses (1988 to 1997) who were prenatally diagnosed with gastroschisis by a sonologist in the Ultrasound Genetic Unit, Department of Obstetrics and Gynecology at Washington University, were reviewed. RESULTS: Oligohydramnios and intrauterine growth restriction were diagnosed in 23% and 49% of the pregnancies, respectively. A total of 22 women underwent induction of labor nine for nonreassuring fetal testing, four for premature rupture of membranes, five for marked bowel dilatation, one for preeclampsia, and three for other reasons. Cesarean section (CS) was performed in 16 of 43 (37%) of women. The indications for CS were fetal distress (9 of 16 women), chorioamnionitis (2 of 16 women), breech presentation (3 of 16 women), and physician discretion (2 of 16 women). No significant differences in Apgar scores were observed between the fetuses. Fetuses who were delivered by CS for fetal distress were more likely to have undergone an induction of labor (91% versus 44%), and they were smaller than fetuses with no evidence of fetal distress (2220 +/- 105 gm versus 2613 +/- 80 gm, p < 0.05). CONCLUSION: The incidence of antepartum and intrapartum complications in fetuses with gastroschisis is high. The rate of CS can reach 37%. These data may aid clinicians in counseling patients with gastroschisis. PMID- 10685304 TI - Parent's grand rounds speech on neonatal intensive care unit experience. PMID- 10685305 TI - Neonatal myiasis: a case report and a role of the Internet. AB - Maggot infestation (myiasis) can occur in the newborn baby. However, neonatal myiasis is rare, and there is no published literature on this subject. Rapid useful information on such an esoteric clinical case can be obtained by searching the Internet. Effective medical management includes complete removal of the maggots and offering reassurance to the distraught parents. PMID- 10685306 TI - "Bloodless" treatment of a Jehovah's Witness infant with ABO hemolytic disease. AB - An ABO-incompatible term infant girl born to parents who are Jehovah's Witnesses was admitted to our neonatal unit with a high serum bilirubin level necessitating exchange transfusion. The parents signed a request that blood should not be administered under any circumstances. However, they authorized us to apply the possible alternative treatments of orally administered D-penicillamine (300 mg/kg per day divided in three doses for 3 days), phototherapy, intravenous fluids, and recombinant human erythropoietin (200 U/kg subcutaneously on every second day for 2 weeks). Herein, we report the outcome of this baby, who was discharged from the our unit in good condition after treatment. Her physical growth and motor milestones at 14 months of age revealed no red flags for neurodevelopmental maturation. To our knowledge, this is the first case of an infant who received such a combined alternative (and "bloodless") treatment of serious ABO hemolytic disease of the newborn. PMID- 10685307 TI - Fatal early onset infection in an extremely low birth weight infant due to Morganella morganii. AB - OBJECTIVE: This paper reports a case of chorioamnionitis due to Morganella morganii in a mother who presented with ruptured membranes at 24 weeks' gestation and was treated with dexamethasone and prophylactic ampicillin. Her premature infant developed severe early onset infection due to the same organism and expired. STUDY DESIGN: A clinical case report of M. morganii infection complicating preterm rupture of membranes is presented. Possible risk factors for maternal and neonatal infection with this organism as well as the therapy of neonatal M. morganii infection are discussed. RESULTS: Risk factors in the mother included having a cervical cerclage in place and treatment with dexamethasone and prophylactic ampicillin. The major risk factors in the infant were maternal chorioamnionitis and extreme prematurity. The mother responded to treatment with ampicillin, metronidazole, and gentamicin following delivery and had an uncomplicated recovery. Her infant developed severe early onset M. morganii infection complicated by neutropenia, thrombocytopenia, and severe acidosis and expired. Postmortem cultures of pleural fluid, peritoneal fluid, and blood were positive despite treatment with gentamicin, an antibiotic to which the organism was sensitive. CONCLUSION: M. morganii may cause serious infection in pregnancy and in the neonatal period. The use of dexamethasone and prophylactic ampicillin may have increased the risk of infection with this ampicillin-resistant organism. The failure of gentamicin to sterilize the infant's blood and body fluids emphasizes the necessity of treating such infections with a combination of an aminoglycoside and a third-generation cephalosporin, such as cefotaxime. PMID- 10685308 TI - Neonatal pyoscrotum and perforated appendicitis. AB - Neonatal appendicitis is so unusual that is has been considered nonexistent. We present the case of a 6 day old male with acute scrotal swelling who was found to have a perforated appendicitis inside a hernia sac. A literature review supports the impression that neonatal appendicitis is rare; however, when it occurs inside a hernia sac, it has a good prognosis. PMID- 10685309 TI - Progressive tumor necrosis and lethal hyperkalemia in a neonate with sacrococcygeal teratoma (SCT). AB - Tumor lysis syndrome is known among patients undergoing induction therapy for lymphocytic malignancies. Spontaneous tumor lysis in patients with solid tumors is distinctly rare. To our knowledge, the phenomenon of spontaneous tumor lysis has been described only once in infancy, in association with the surgical manipulation of a hepatoblastoma. This is the first report of a newborn with sacrococcygeal teratoma who experienced spontaneous tumor lysis-induced hyperkalemia. Because cardiac arrest may be among the leading causes of operative mortality in babies with sacrococcygeal teratoma, intraoperative monitoring of serum K+ should be conducted frequently. PMID- 10685310 TI - Fetal heart rate monitoring casebook. Checkmark fetal heart rate pattern and normal neonatal outcome. AB - BACKGROUND: A checkmark pattern of the fetal heart rate (FHR) had been seen in association with hypoxia in human and animal fetuses. CASE: We report a checkmark pattern in a fetus of a 26-year-old primigravida who underwent induction of labor at term. Scalp pH was 7.32. Comprehensive ultrasound examination revealed normal results. The neonate was born in satisfactory condition. Apgar scores were 7 and 10 at 1 at 5 minutes of life, respectively. CONCLUSION: Checkmark FHR pattern may be a benign finding; however, if it is detected, it should be closely monitored. PMID- 10685311 TI - Special imaging casebook. Neonate with osteogenesis imperfecta and asplenia syndrome with horseshoe adrenal. PMID- 10685312 TI - Recommended standards for newborn ICU design. Committee to establish recommended standards for newborn ICU design. PMID- 10685313 TI - Condoms. PMID- 10685314 TI - Breakage and slippage of condoms among users in north Gondar Province, Ethiopia. AB - OBJECTIVE: To determine the frequency of condom breakage and slippage among male condom users and to analyse some factors affecting condom breakage. DESIGN: Prospective study undertaken from March to June 1996. SETTING: Gondar, Teda and Aykel health centres in north Gondar province. PARTICIPANTS: One hundred and seventy literate male condom users who volunteered to participate in the study were enrolled, but data were collected and analysed for 143 persons. MAIN OUTCOME MEASURES: Condom breakage was measured as the number of condom broken while opening the package, putting on, during intercourse or withdrawal; and condom slippage was measured as the number of condoms slipped off the penis during intercourse or withdrawal. RESULTS: Of 143 participants, 26.6% broke condoms. Total condom breakage rate was 4.6% and slippage rate was 0.1%; half of the condom breakage occurred at the tip. Condom breakage was more observed in less educated (p < 0.05). Breakage rate and experience of condom use were negatively correlated (r = -0.214, P < 0.05). CONCLUSION: User's knowledge and experience reduce the breakage of condom. PMID- 10685315 TI - Predictors of women's decision to ask new partners to use condoms to avoid HIV/AIDS in South Africa. AB - OBJECTIVES: (i) to identify the key determinants of women's decision to ask a new partner to use condoms during sexual intercourse in order to reduce the probability of HIV infection. (ii) to estimate the impact of social, economic and educational factors identified above on the likelihood of a positive decision and; (iii) to explore the policy implications of the analysis. DESIGN: Cross sectional national household sample survey. SETTING: South African Health Inequalities Survey, 1994. SUBJECTS OR PARTICIPANTS: Three thousand three hundred and thirty three South African women aged between 16 and 64 years. INTERVENTIONS: Descriptive/inferential non-intervention study. MAIN OUTCOME MEASURES: Respondent's decision to ask one's new partner to use a condom during sexual intercourse. RESULTS: The study revealed that condoms have a contraceptive benefit; AIDS epidemic is spreading rapidly across South Africa; people in ones own community are using condoms to avoid getting AIDS; and AIDS can be transmitted by having sexual intercourse with someone without using a condom were positive and statistically significant at p < or = 0.05. CONCLUSION: Equipping women with above mentioned forms of knowledge would go a long way in empowering them to exercise their rights to uncoerced choice to have safe sexual relationships. PMID- 10685316 TI - Pregnancy outcome in primigravidae with late onset hypertensive disease. AB - BACKGROUND: The perinatal mortality associated with pre-eclampsia is extremely high and it is mainly associated with early onset disease in multiparous women. Hypertension without proteinuria in late pregnancy may not be associated with high perinatal mortality rates. OBJECTIVE: To establish the perinatal outcome in primigravidae women with hypertension occurring in late pregnancy, that is, at thirty fourth week or later. DESIGN: Prospective case-control study. SETTING: Labour ward of King Edward VIII Hospital, Durban, South Africa. PATIENTS: Three hundred and twenty two primigravidae consisting of 161 hypertensives and 161 controls. MAIN OUTCOME MEASURES: Maternal and foetal morbidity and mortality. RESULTS: The hypertensive group was divided into those with proteinuria (group a) and without proteinuria (group b). The mean birthweight of babies born to proteinuric hypertensives was significantly lower than that of hypertensives without proteinuria and the normotensive group (2.4 kg (a) versus 2.8 kg (b) versus 3.02 kg (c) respectively--a versus b, p = 0.0001; a versus c, p = 0.001; b versus c, p = 0.009). There were nine perinatal deaths and all occurred in the proteinuric hypertension group. CONCLUSION: Primigravidae with late onset proteinuric hypertension had smaller babies and higher perinatal mortality than their aproteinuric hypertensive and normotensive controls. PMID- 10685317 TI - Demographic and socio-economic factors with implications for the control of lymphatic filariasis in Kwale District, Kenya. AB - OBJECTIVE: To examine demographic and socio-economic factors with regard to control of lymphatic filariasis. DESIGN: A cross sectional data collection approach using a questionnaire. SETTING: Three adjacent villages, namely Lutsangani, Dzivani and Gandini in Kwale district. This was the study site for a larger KEMRI-JICA lymphatic filariasis control project. Findings from the larger project showed filariasis prevalence rates of 22.6%, 12.1% and 11.9% in the three villages respectively. SUBJECTS: All 891 household-heads in the three villages. MAIN OUTCOME MEASURES: Relationship between selected factors (age, sex, education, employment status, availability of latrine and permanent roofing for the main dwelling) and current use of protection against mosquito bites. RESULTS: Use of mosquito-bed nets, mosquito repellents and insecticides was found in 13.8% of the households. Education and employment status of the household-heads were positively associated with protection against man-vector contact (p < 0.001). Non availability of latrines and lack of permanent roofing (tiles or corrugated iron sheets) for the main dwellings in 80% and 95% households, respectively, were additional clear indicators of low socio-economic development. CONCLUSION: Given the inter-village variations in the prevalence of the filariasis, wide scale diagnosis for treatment control approach could be prohibitively expensive. Effective control of filariasis in this and similar communities may be by mass treatment with either diethylcarbamazine (DEC) or ivermectin combined with albendazole. PMID- 10685319 TI - Re: Cutaneous leishmaniasis in BALB/c mice caused by a sauroleishmania species isolated from a plated lizard, gerrhosaurus major (squamata cordylidae) PMID- 10685318 TI - Incarcerated and strangulated inguinal hernias in children in Zaria, Nigeria. AB - OBJECTIVE: To reappraise the problem of incarcerated and strangulated inguinal hernias in children in Zaria, Nigeria. DESIGN: A retrospective study. SETTING: Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. PATIENTS: Eighteen children aged below two years with incarcerated and strangulated inguinal hernias managed over a 10 year period. MAIN OUTCOME MEASURES: Incarceration and strangulation rate, resection rate, testicular infarction, wound infection. RESULTS: The overall incarceration and strangulation rate was 4.8%. The rate in neonates was 80%, 33% in those less than six months and 21% in those below one year. There was no incarceration or strangulations after two years of age. The intestinal resection rate for gangrene was 11% (two neonates) and ipsilateral orchidectomy was necessary for testicular infarction in two neonates (11%). Wound infection occurred in three patients (17%) and there were no deaths. Overall, there has been an improvement in the incarceration and strangulation rate, resection rate and wound infection rate over earlier years (5.7%, 28% and 50% respectively for earlier years) due largely to early presentation. Testicular infarction, however, remains a major risk particularly in neonates. CONCLUSION: The policy of early repair of inguinal hernias in children especially below two years and particularly in neonates should be continuously emphasised to avoid morbidity. PMID- 10685320 TI - Correlates of psychiatric morbidity and case identification in Ibadan, Nigeria. AB - OBJECTIVE: To determine the correlates of psychiatric morbidity and the factors that may influence the recognition of psychiatric cases in primary care. DESIGN: In the cross-sectional survey, the sample was selected by systematic sampling technique. Each subject was interviewed in sequence by a research nurse, a psychiatrist and a general practitioner (GP). SETTING: The primary care facility at the University College Hospital, Ibadan. SUBJECTS: Eight GPs and 414 (44% males, 56% females) patients were selected. INTERVENTION: A socio-demographic questionnaire, the present state examination (PSE), and clinical interview were administered to the subjects. Eysench personality questionnaire (EPQ) was completed by GPs. The research nurses scored each GP on an enthusiasm scale. MAIN OUTCOME MEASURES: The diagnosis by psychiatrist and GPs recorded on doctors' information sheet: the PSE scores of subjects, the EPQ scores of GPs and their observed enthusiasm ratings among others were the main measures. RESULTS: The psychiatric morbidity rate was 43.4% in the centre. GPs accuracy was 0.7 and Kappa score was 0.44. EPQ Neuroticism and Psychoticism scores correlated negatively with GPs ability to recognise cases, whereas enthusiasm correlated positively. GPs had difficulty in recognising minor psychiatric disorders accounting for 94.4% of morbidity in the primary care. Morbidity was significantly associated with female sex, age 45-54 years, self-employment, low level of education and being currently married. Females and the self-employed were over-represented among GPs' cases. CONCLUSION: More data from similar studies across cultures would provide useful insight into strategies that could bring about early recognition and treatment of minor psychiatric disorders in the primary health care. PMID- 10685321 TI - Nutrition in the management of acquired immunodeficiency syndrome. AB - OBJECTIVE: To highlight the influence of nutrition on the progress of HIV/AIDS and the role and importance of good nutrition in the management of the disease. STUDY SELECTION: The subject was selected because it is now recognised that nutritional care and support is an essential component of the health care plan and management for people with HIV/AIDS. The subject is especially relevant since few studies have been conducted locally on the effects of nutritional status on the progression of HIV/AIDS. DATA RESOURCES AND DATA SYNTHESIS: A review of current literature selected from local and international scientific journals and books on the subject of nutrition and HIV/AIDS. DATA EXTRACTION AND SYNTHESIS: Data were developed from the reviewed information extracted from the contribution of different authors who are interested in nutritional management of people with HIV/AIDS. It was then analysed and synthesised into the current article. CONCLUSION: On the basis of the reviewed information, it is recommended that individuals with HIV/AIDS be given nutrition counselling and support to enable them achieve an adequate nutrient and energy intake for as long as possible. This would enhance the quality of their lives and minimise disease symptoms. PMID- 10685322 TI - Nutritional status and nutrient intake of preschool children in northern Ghana. AB - OBJECTIVE: To determine the nutritional status and nutrient intake of preschool children in a sub-Sahelian setting so as to ascertain whether they meet the acceptable standards. DESIGN: Prospective/investigative study. SETTING: The study was carried out in Saboba, a rural sub-Sahelian village in northern Ghana. SUBJECTS: Five hundred and eighteen preschool children (2.5-6 years) drawn from three kindergartens and five other localities were used. INTERVENTION: Anthropometric measures of age, weight, and height were assessed. Blood and stool samples were taken for analyses. RESULTS: The nutritional status was generally poor, with 27% (140/519) stunted, 4.4% (23/519) wasted and 1.9% (10/519) wasted and stunted. Majority of the children (92%) were anaemic and 16.3% had deficient level of vitamin A (< 10 micrograms/dL). Total caloric intake was low (only 26.5% met RDA values) and apart from protein and iron whose RDA were met by the majority of the children (> 90%), the intake of other nutrients-calcium, vitamin C, vitamin A and provitamin A, were low and unacceptable, compared to their respective RDA values. In the case of iron, even though intake was adequate, the high anaemic rate of 92% was due to high incidence of malaria (32%), hookworm infestation, low intake of vitamin C and low bioavailability of iron from the mainly cereal diet. CONCLUSION: The poor nutritional status of the children was largely due to the low intake of essential nutrients. PMID- 10685323 TI - Magnitude and determinants of bottle feeding in rural communities. AB - BACKGROUND: Though bottle feeding is known to be hazardous either due to over dilution of the supplement or faulty hygienic techniques during its preparation, especially in areas where general sanitation is poor, very little attention has been paid to it in developing countries. OBJECTIVE: To determine the extent of bottle use in child feeding and the factors associated with its use in rural communities. DESIGN: Community based cross-sectional study. PATIENTS: One thousand five hundred and thirty six children, aged 0-23 months and their mothers were included in the study. RESULTS: The overall prevalence of bottle feeding was 11.3%. Out of the 174 children who were bottle fed, only 11(6.3%) were exclusively on it. Residence, maternal education and occupation were significantly associated with the practice of bottle feeding in the crude analysis and after adjusting for parental and child characteristics (p < 0.05). CONCLUSION: The extent of bottle feeding in the studied communities is generally high, with a higher rate among town women, a trend which was seen in the developed world at the beginning of the century. Improvement in maternal and child health services including education on child feeding are recommended. PMID- 10685324 TI - Iron deficiency anaemia in children of a peri-urban health facility. AB - OBJECTIVE: To ascertain the prevalence of iron deficiency anaemia(IDA) and its risk factors. DESIGN: A cross-sectional survey. SETTING: A peri-urban health centre in Nairobi, Kenya. SUBJECTS: Four hundred and three children, aged six months to six years. INTERVENTION: Demographic data were obtained and each child examined for signs of iron deficiency anaemia. Blood was drawn for haemoglobin determination. MAIN OUTCOME MEASURE: The diagnosis of iron deficiency anaemia was made using a pre-defined criteria. RESULTS: Iron deficiency anaemia had a prevalence of 7.4% (95% CI = 4.8-10.0) and was predominantly mild (93.6%). Age was found to be significantly associated with iron deficiency anaemia with a prevalence of (14.6%) in infants. No association was found between IDA and sex, birthweight, weaning age and weaning diet, sanitation, water source or mother's education. CONCLUSION: The prevalence of iron deficiency anaemia in this health facility was relatively low and was predominantly mild. PMID- 10685325 TI - Serum eosinophil cationic protein as a predictor of disease activity in acute and chronic asthma. AB - BACKGROUND: Eosinophils may contribute to airway hyper responsiveness in asthma through the effects of eosinophil derived granular proteins in the bronchial epithelium. Increased concentration of eosinophil cationic protein (ECP) has been reported in patients with acute and chronic asthma. OBJECTIVE: To examine if ECP can serve as a marker of disease activity in acute and chronic asthma patients. DESIGN: Prospective case control study. PATIENTS: Sixteen non smoking asthmatics in exacerbation (group 1); twenty two in relatively stable state (group 2); and sixteen normal control subjects (group 3) were recruited into the study. SETTING: Casuality and outpatients departments, Mubarak hospital, Kuwait between August 1997 and July 1998. MAIN OUTCOME MEASURES: The mean serum ECP, blood eosinophil count and peak expiratory flow rate (PEFR). RESULTS: There was a statistically significant difference between the groups in blood eosinophil count (p < 0.01) and in PEFR (p < 0.0001). At week four, the mean ECP and blood eosinophil count fell as a result of therapy in group 1. The difference in PEFR values between week 0 and 4 in group 1 reached statistical significance (p < 0.05). In group 2 patients, the mean serum ECP, blood eosinophil count and PFER values between week 0 and 4 did not show any significant difference. A correlation was observed between ECP and PEFR in group 1 (p < 0.05) and between ECP and eosinophil count in group 2 (p < 0.01). CONCLUSION: Serum ECP has the potential to serve as a marker for predicting and monitoring the clinical course of asthma. Further studies are required to verify these baseline findings in our environment. PMID- 10685326 TI - KEMRI Hep-cell II hepatitis B surface antigen screening kit. AB - BACKGROUND: Kenya is a high hepatitis B virus (HBV) endemic zone. Prevention of HBV transmission by transfusing safe blood is necessary. Kits for screening hepatitis B surface antigen (HBsAg) are usually imported and are expensive. Hence it has been difficult to screen donated and patient blood samples all over Kenya. OBJECTIVE: To produce a HBsAg screening kit locally in order to be able to screen donated and patient blood samples all over Kenya. DESIGN: A laboratory based study. SETTING: Centre for Virus Research (CVR), Kenya Medical Research Institute (KEMRI), Nairobi. METHOD: Purified HBsAg from plasma of carriers obtained from National Public Health Laboratories Services (NPHLS) was used to minimise guinea pigs to produce antihepatitis B (anti HBs) antibody. The anti HBs was then used to sensitise sheep red blood cells (SRBC). The final product was freeze dried (lyophilised) and its sensitivity and specificity was compared with other commercial kits. RESULTS: The sensitivity and specificity of KEMRI Hep-cell II was found to be 98% and 99%, respectively. The kit was found to be stable and potent for one year whether kept 4 degrees C, 37 degrees C or room temperature. CONCLUSION: KEMRI Hep-cell II was successfully produced locally. The sensitivity and specificity were comparable to other commercial kits. The kit was stable and potent for one year between temperature of 4 degrees C and 37 degrees C. The kit required only simple apparatus to carry out the test hence it can be used anywhere in Kenya. It was also cheap and affordable. PMID- 10685327 TI - Small bowel and mesenteric injury following traditional treatment and self inflicted trauma to inguinal hernia. AB - Inguinal hernia is a common indication for emergency surgery in our environment after intestinal obstruction. Occasionally other rare indications may be seen. Recently, two patients with such unusual presentation were managed in this institution. One had his hernia incised, by a traditional barber with evisceration and the other incised his hernia himself with evisceration and laceration of the small bowel. Both patients were successfully managed. The menace of traditional barbers and medical ignorance in tropical Africa are reported. PMID- 10685328 TI - Hypertrophic cardiomyopathy: case report. AB - A case report of a 19 year old young adult male who died suddenly without any apparent clinical cause is presented. Autopsy examination revealed hypertrophied interventricular septum and left ventricle suggestive of a hypertrophic cardiomyopathy (HCM). About 50% of hypertrophic cardiomyopathies are inherited as autosomal dominant disorder, sometimes associated with neuroectodermal syndromes, such as neurofibromatosis and pheochromocytoma. HCM is one of the common causes of severe arrhythmias and sudden death. PMID- 10685329 TI - Does female age affect embryo morphology? AB - Deteriorating oocyte quality is commonly believed to be the primary determinant of the decreased implantation potential in older women. We assessed the influence of age on embryo morphology in standard in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) modalities. All 6350 consecutive embryos (2990 IVF, 3360 ICSI) obtained in our Assisted Reproductive Technology Unit from January 1996 through June 1997 were included. High quality embryos were defined as those with equal-sized blastomeres and < 10% fragmentations and a cleavage rate of four cells on day 2 or eight cells on day 3 transfers. The results were analyzed for the standard IVF group, the ICSI group, and the ICSI subgroup with severe male factor infertility (< or = 1 x 10(6) total motile spermatozoa in the ejaculate). For standard IVF, a positive association was observed between female age and increased proportion of good quality embryos. No such association was detected for the ICSI cycles (whole group or subgroup). We conclude that in standard IVF, embryo quality, as reflected by embryo morphology, does not deteriorate with increased maternal age. PMID- 10685330 TI - Follow-up of 32 hypothalamo-hypopituitary patients treated with pulsatile gonadotropin-releasing hormone or human menopausal gonadotropin. AB - In a clinical retrospective study, a follow-up of hypothalamo-amenorrheic patients treated firstly with gonadotropin-releasing hormone (GnRH) pump stimulation and secondly with human menopausal gonadotropin (hMG) was performed. Thirty-two hypothalamo-amenorrheic patients, 24-38 years old, were submitted to 103 GnRH stimulation cycles. Seven, with polycystic ovaries (PCO) on ultrasound, were stimulated with hMG after one or several unsuccessful pump cycles. Ovulation was confirmed by a luteinizing hormone (LH) surge or triggered by human chorionic gonadotropin in 80 out of 103 cycles (77.7%/cycle) leading to 62 timed sexual intercourses and 17 intrauterine inseminations (IUI). Twenty-one pregnancies (26.3%/cycle) terminated in eight abortions (38.1%/pregnancy) and 13 deliveries (40.6%/patient). hMG stimulation, in the seven PCO patients (six IVF, one IUI), led to four additional deliveries in three patients. Five patients became pregnant spontaneously after pump failure (n = 2) or unsuccessful IVF (n = 3). Combining all cycles, 17 deliveries were obtained in 16 patients. No case of ovarian hyperstimulation syndrome (OHSS) was observed. GnRH is an efficient and safe treatment of hypothalamo-amenorrheic-induced anovulation. Following GnRH or hMG ovarian stimulation, spontaneous ovulation and conception may be restored in certain hypothalamo-amenorrheic patients. PMID- 10685331 TI - Estriol add-back therapy in the long-acting gonadotropin-releasing hormone agonist treatment of uterine leiomyomata. AB - The hypoestrogenic state induced by gonadotropin-releasing hormone agonists (GnRHa) has been shown to be effective in the treatment of uterine leiomyomas but to induce bone loss. Estriol has been described to be a weak and short-acting estrogen without an increased risk of endometrial proliferation and hyperplasia. The purpose of this study was to evaluate whether treatment of uterine leiomyomata with GnRHa plus oral estriol add-back therapy could prevent bone loss, without deteriorating the therapeutic effect of GnRHa. Twelve premenopausal women with symptomatic uterine leiomyomas were randomized to receive either leuprolide acetate depot alone at a dose of 3.75 mg s.c. every month for 6 months (non add-back group; n = 6), or GnRHa for 6 months plus oral estriol 4 mg/day for 4 months commencing with the third GnRHa injection (add-back group; n = 6). In the add-back group, leiomyoma volume, as measured by transvaginal ultrasound, decreased to 59.1% of baseline at 2 months of GnRHa therapy with no significant change in size during the remaining treatment period. In contrast, it decreased to 31.3% of pretreatment size at the end of treatment in the non add-back group. The levels of bone metabolic markers such as CrossLaps, deoxypyridinoline, osteocalcin and bone-specific alkaline phosphatase, increased significantly throughout the treatment in the non add-back group, whereas they were suppressed by the add-back therapy. The bone mineral density of lumbar spine (L2-L4) as measured by dual-energy X-ray absorptiometry decreased significantly by 7.5% at the end of treatment in the non add-back group, but did not change significantly in the add-back group. In conclusion, GnRHa plus estriol add-back therapy might be considered for long-term treatment of uterine leiomyomata. PMID- 10685332 TI - Prevalence of a positive family history of type 2 diabetes in women with polycystic ovarian disease. AB - The known association between insulin resistance and polycystic ovarian disease (PCOD) has been studied by determination of the prevalence of a positive family history of diabetes in a consecutive series of oligomenorrheic women with polycystic ovaries and eumenorrheic women with normal ovaries who served as controls. A significantly greater proportion of the families of the patients with PCOD had at least one member affected by type 2 diabetes (39.1% of the PCOD group and 7.6% of the controls; p < 0.001). Both obese (54.8%) and non-obese women (24.2%) with PCOD had an increased prevalence of type 2 diabetes within their families. Paternal and maternal family members affected were in similar proportions, there being no evidence of preferential transmission through the female line in this study. The increased prevalence of type 2 diabetes in the families of women with polycystic ovaries is further evidence for the association between PCOD and insulin resistance, and provides a possible explanation for the familial nature of the ovarian disorder. PMID- 10685333 TI - Total testosterone and DHEAS levels as predictors of androgen-secreting neoplasms: a populational study. AB - Androgen excess affects between 2% and 10% of women. While the majority of these patients suffer from polycystic ovary syndrome, a few present with an androgen secreting neoplasm. An elevated circulating total testosterone level and dehydroepiandiosterone sulfate (DHEAS) level have been proposed as screening methods for detecting ovarian and adrenal androgen-secreting neoplasms, respectively. To determine the predictive value of these tests for androgen secreting tumors in a population of consecutive hyperandrogenic patients, we studied 478 consecutive untreated hyperandrogenic patients presenting over a ten year period (1987-97). All had at least two of the following features: (1) oligomenorrhea (i.e. cycles > 35 days or < 8 cycles/year), (2) hyperandrogenemia (i.e. a total or free testosterone, or DHEAS > 95th percentile of controls), or (3) hirsutism (i.e. a modified Ferriman-Gallwey score > or = 6). None of these patients had a prior diagnosis of an androgen-secreting neoplasm. Basal levels of testosterone and DHEAS were determined in all patients, with transvaginal sonography and an adrenal computed tomography scan in select individuals. Of the 478 patients included, 65% had hirsutism and oligomenorrhea; 20% had hyperandrogenic oligomenorrhea; and 15% had hirsutism and hyperandrogenemia, without overt oligomenorrhea. Overall, 11 (2.3%) patients had a total testosterone > 8.7 nmol/l (250 ng/dl), of which one actually had an androgen secreting neoplasm (i.e. true-positive). This postmenopausal patient presented with rapidly progressive virilization, and demonstrated an ovarian hilar cell tumor at surgery. The calculated sensitivity of an elevated testosterone level (> 8.67 nmol/l) for a neoplasm was 100% (1/1), the specificity was 98% (467/477), and the negative predictive value was 100% (467/467), but the positive predictive value was only 9% (1/11). Ten subjects had DHEAS levels > 16.3 mumol/l (6000 ng/ml), and none was diagnosed with an adrenocortical tumor. Although the sensitivity and positive predictive value of a high DHEAS for a neoplasm could not be calculated due to the absence of a test case, the specificity was 98% (468/478) and the negative predictive value was 100% (468/468). These data suggest that the measurement of testosterone and DHEAS is not a cost-effective method of screening for these tumors, due to the low frequency of the disorder and the fact that clinical evaluation alone is often sufficient screening. PMID- 10685334 TI - LH-RH analogues: I. Their impact on reproductive medicine. AB - In the 28 years that have passed since the elucidation of the structure of luteinizing hormone-releasing hormone (LH-RH), diverse clinical applications in the field of reproductive medicine and related fields have been established for agonists of LH-RH, based on inhibition of the pituitary-gonadal axis. Various clinical investigations with agonists of LH-RH in benign gynecologic disorders, polycystic ovary disease (PCOD), in vitro fertilization-embryo transfer (IVF-ET), benign prostatic hypertrophy (BPH), precocious puberty and contraception were reviewed. LH-RH antagonists inhibit LH, follicle-stimulating hormone (FSH), and sex steroid secretion immediately after their administration and thus achieve rapid therapeutic effects. LH-RH antagonists should find applications in the treatment of uterine leiomyomas, endometriosis, and in controlled ovarian stimulation-assisted reproductive techniques (COS-ART), which have been already established for the agonists. Modern LH-RH antagonists such as cetrorelix may prove superior to the agonists in COS-ART and also in the treatment of BPH, but additional studies in these and other areas are needed. PMID- 10685335 TI - Regulation of granulosa cell proliferation and apoptosis during follicular development. PMID- 10685336 TI - A historical perspective of the clinical evolution of the assisted reproductive technologies. AB - The practice of assisted reproduction technology today is the result of the dedicated patient care, observation, research, and experimentation undertaken by previous generations of physicians. The building blocks of progress have been assembled over past decades, by scientists whose primary objective has been to push forward the frontiers of knowledge, in order to offer more effective methods of infertility treatment. And fortunately that process continues today. Amongst the many scientific developments that have led to the modern practice in assisted reproductive technology, a small number stand out as having had a unique importance. This historical review redraws the path through which in vitro fertilization went from an experimental to an accepted infertility treatment. PMID- 10685337 TI - Endocrine therapy for gynecological cancer. PMID- 10685339 TI - Multiple-quantum HCN-CCH-TOCSY experiment for 13C/15N labeled RNA oligonucleotides. AB - A multiple-quantum 3D HCN-CCH-TOCSY experiment is presented for the assignment of RNA ribose resonances. The experiment makes use of the chemical shift dispersion of N1 of pyrimidine and N9 of purine to distinguish the ribose spin systems. It provides an alternative approach for the assignment of ribose resonances to the currently used COSY- and TOCSY-type experiments in which either 13C or 1H is utilized to distinguish the different spin systems. Compared to the single quantum version, the sensitivity of the multiple-quantum HCN-CCH-TOCSY experiment is enhanced on average by a factor of 2 for a 23-mer RNA aptamer complexed with neomycin. PMID- 10685338 TI - Refined solution structure and backbone dynamics of 15N-labeled C12A-p8MTCP1 studied by NMR relaxation. AB - MTCP1 (for Mature-T-Cell Proliferation) was the first gene unequivocally identified in the group of uncommon leukemias with a mature phenotype. The three dimensional solution structure of the human p8MTCP1 protein encoded by the MTCP1 oncogene has been previously determined by homonuclear proton two-dimensional NMR methods at 600 MHz: it consists of an original scaffold comprising three alpha helices, associated with a new cysteine motif. Two of the helices are covalently paired by two disulfide bridges, forming an alpha-hairpin which resembles an antiparallel coiled-coil. The third helix is orientated roughly parallel to the plane defined by the alpha-antiparallel motif and appears less well defined. In order to gain more insight into the details of this new scaffold, we uniformly labeled with nitrogen-15 a mutant of this protein (C12A-p8MTCP1) in which the unbound cysteine at position 12 has been replaced by an alanine residue, thus allowing reproducibly high yields of recombinant protein. The refined structure benefits from 211 additional NOEs, extracted from 15N-edited 3D experiments, and from a nearly complete set of phi angular restraints allowing the estimation of the helical content of the structured part of the protein. Moreover, measurements of 15N spin relaxation times and heteronuclear 15N?1H?NOEs provided additional insights into the dynamics of the protein backbone. The analysis of the linear correlation between J(0) and J(omega) was used to interpret relaxation parameters. It appears that the apparent relative disorder seen in helix III is not simply due to a lack of experimental constraints, but associated with substantial contributions of sub-nanosecond motions in this segment. PMID- 10685340 TI - Maximum entropy approach to the determination of solution conformation of flexible polypeptides by global conformational analysis and NMR spectroscopy- application to DNS1-c-[D-A2,bu2,Trp4,Leu5]enkephalin and DNS1-c-[D-A2bu2,Trp4,D Leu5]enkephalin. AB - A method is proposed to determine the conformational equilibrium of flexible polypeptides in solution, using the data provided by NMR spectroscopy and theoretical conformational calculations. The algorithm consists of the following three steps: (i) search of the conformational space in order to find conformations with reasonably low energy; (ii) simulation of the NOE spectrum and vicinal coupling constants for each of the low energy conformations; and (iii) determining the statistical weights of the conformations, by means of the maximum entropy method, in order to obtain the best fit of the averaged NOE intensities and coupling constants to the experimental quantities. The method has been applied to two cyclic enkephalin analogs: DNS1-c-[D-A2bu2,Trp4,Leu5]enkephalin (ENKL) and DNS1-c-[D-A2bu2,Trp4,D-Leu5]enkephalin (ENKD). NMR measurements were carried out in deuterated dimethyl sulfoxide. Two techniques were used in conformational search: the electrostatically driven Monte Carlo method (EDMC), which results in extensive search of the conformational space, but gives only energy minima, and the molecular dynamics method (MD), which results in a more accurate, but also more confined search. In the case of EDMC calculations, conformational energy was evaluated using the ECEPP/3 force field augmented with the SRFOPT solvation-shell model, while in the case of MD the AMBER force field was used with explicit solvent molecules. Both searches and subsequent fitting of conformational weights to NMR data resulted in similar conformations of the cyclic part of the peptides studied. For both ENKL and ENKD a common feature of the low-energy solution conformations is the presence of a type II' or type IV beta-turn at residues 3 and 4; the ECEPP/3 force field also gives a remarkable content of type III beta-turn. These beta-turns are tighter in the case of ENKL, which is reflected in different distributions of the D-A2bu(N gamma H)...D A2bu(CO) and D-A2bu(N gamma H)...Gly3(CO) hydrogen-bonding distances, indicating that the D-A2bu(N gamma H) amide proton is more shielded from the solvent than in the case of ENKD. This finding conforms with the results of temperature coefficient data of the D-A2bu(N gamma H) proton. It has also been found that direct (MD) or Boltzmann (EDMC) averages of the observables do not exactly conform with the measured values, even when explicit solvent molecules are included. This suggests that improving force-field parameters might be necessary in order to obtain reliable conformational ensembles in computer simulations, without the aid of experimental data. PMID- 10685342 TI - 1H, 15N, 13C, and 13CO assignments and secondary structure determination of RGS4. PMID- 10685341 TI - Line narrowing in spectra of proteins dissolved in a dilute liquid crystalline phase by band-selective adiabatic decoupling: application to 1HN-15N residual dipolar coupling measurements. AB - Residual heteronuclear dipolar couplings obtained from partially oriented protein samples can provide unique NMR constraints for protein structure determination. However, partial orientation of protein samples also causes severe 1H line broadening resulting from residual 1H-1H dipolar couplings. In this communication we show that band-selective 1H homonuclear decoupling during data acquisition is an efficient way to suppress residual 1H-1H dipolar couplings, resulting in spectra that are still amenable to solution NMR analysis, even with high degrees of alignment. As an example, we present a novel experiment with improved sensitivity for the measurement of one-bond 1HN-15N residual dipolar couplings in a protein sample dissolved in magnetically aligned liquid crystalline bicelles. PMID- 10685343 TI - Backbone NMR assignment and secondary structure of ribosome recycling factor (RRF) from Pseudomonas aeruginosa. PMID- 10685344 TI - Sequence-specific 1H, 13C and 15N resonance assignments of recombinant onconase/P 30 protein. PMID- 10685345 TI - 1H, 15N, and 13C NMR backbone assignments of the N-terminal region of human erythrocyte alpha spectrin including one structural domain. PMID- 10685346 TI - Toward progress research: closing the gap between family therapy practice and research. PMID- 10685347 TI - A content analysis of research in family therapy journals. AB - In order to assess trends in family therapy research, empirical articles (N = 195) from three family therapy journals over a 5-year period were coded for several variables: authorship, external funding, methodology, sample, purpose, cost effectiveness, use of therapeutic model, and topic. Results indicated that a large pecentage of research in these journals focused on nonclinical issues and used nonclinical samples. Authors were affiliated with a wide variety of disciplines and reported low levels of external funding for their research. While a majority of the studies used quantitative methods, there appeared to be a growing number of studies using qualitative methods. Implications of these findngs are discussed in light of research reviews over the past two decades. PMID- 10685348 TI - The use of theory in family therapy research: a content analysis of family therapy journals. AB - Ninety-five studies from Family Process and Journal of Marital and Family Therapy were evaluated with regard to their use of theory. While a majority of the articles were judged to use theory in either an explicit or an implicit manner, 42% did not appear to draw on theory in either the introductory or discussion sections. Studies that used qualitative methods appeared to use theory more frequently and explicitly than those using quantitative methods alone. Systems theory was found to be the most common conceptual framework, followed by feminism. We conclude that the link between theory and research in family therapy needs strengthening and suggest that the role of theory in family therapy be reexamined. PMID- 10685349 TI - The "coming of age" of couple therapy: a decade review. AB - This article overviews significant developments in couple therapy over the last decade. Key trends include: (1) couple therapy becoming firmly established as the accepted treatment of choice for couple problems, (2) the blossoming of the science of relationships, (3) strong evidence supporting the effectiveness of couple therapy both for relationship problems and DSM disorders, (4) greater understanding of the ramifications of gender, (5) new respect for the diversity of family forms, (6) increased accent on the role of emotion, (7) the influence of postmodernism, (8) greater recognition of couple violence, and (9) the move toward integration across models of treatment. PMID- 10685350 TI - Qualitative evaluation of family therapy programs: a participatory approach. AB - In this article, we provide examples of participatory qualitative evaluation methods that can be used within family therapy training programs. These methods can elicit useful feedback to improve programs and, at the same time, empower trainees to become partners in the evaluation process. PMID- 10685351 TI - Recruiting the next generation of marriage and family therapists through undergraduate internships. PMID- 10685352 TI - The intertwined relationship between depression and marital distress: elements of marital therapy conductive to effective treatment outcome. AB - Within the framework of the interpersonal view of depression, this article examines the linkage between depression and four aspects of marital relationship: stress, support, role expectations, and interactional dynamics. Acknowledging the intertwined relationship between depression and marital adjustment, existing models of martial therapy for married depressed patients are examined for the extent to which they address these four aspects. The empirical evidence for the efficacy of these models is also reviewed, suggesting elements of marital therapy that are conducive to effective treatment outcome. PMID- 10685353 TI - A randomized trial of emotion-focused therapy for couples in a training clinic. AB - Forty married couples participated in a randomized trial comparing 8 weekly sessions of emotion-focused therapy (EFT) for couples to a group of couples who were placed on an 8-week waiting list. A composite marital satisfaction score was created from scores on the Dyadic Adjustment Scale, Positive Feelings Questionnaire, and Personal Assessment of Intimacy in Relationships scale. Controlling for pretest scores, participants in the treatment group had significantly higher levels of marital satisfaction after 8 weeks than wait-list participants. Supplementary analyses identified variables associated with gains in therapy and with dropping out of the study. PMID- 10685354 TI - Family therapy trainees' evaluations of their best and worst supervision experiences. AB - Supervisees in Commission on Accreditation for Marriage and Family Therapy Education-(COAMFTE) accredited and candidacy training programs were asked to describe their best and worst supervision experiences in terms of the context of training and supervision, the perceived personal attributes of the supervisor, and specific behaviors perceived to be enacted by the supervisor during supervision. Contextual factors such as supervision modality, frequency and duration of supervisory contacts, and sources of supervisory data appeared to distinguish between supervisees' perceptions of their best and worst experiences. These experiences were further distinguished by the supervisees' perceptions regarding the supervisor's level of interpersonal attractiveness, trustworthiness, and expertise. Key behaviors perceived to be enacted by supervisors during best and worst experiences clustered into several dimensions that included creating an open supervisory environment, communication and encouragement, attending to personal growth, and providing conceptual and technical guidance and direction. PMID- 10685355 TI - Promoting reconciliation through psychoeducational and therapeutic interventions. AB - This article presents a conceptualization of reconciliation as an interpersonal process that can mend relationships in which trust has been violated. We define reconciliation and describe a six-step program for promoting explicit reconciliation in couples. Psychoeducational groups of couples and couple therapy are often the contexts for promoting reconciliation. We discuss some theoretical support for each step, techniques used to promote progress through the step, and differential considerations in psychoeducation and therapy. PMID- 10685356 TI - Training issues for supervisors of marriage and family therapists working with persons living with HIV. AB - The purpose of this article is to address the special issues and considerations Martial and Family Therapy (MFT) supervisors might face with the increasing HIV/AIDS epidemic. Three primary issues will be addressed in this article. First, the importance of educating therapists regarding various aspects of the disease process and its transmission will be discussed, followed by educational strategies programs might adopt. Second, we will discuss the ethical and legal considerations that may need monitoring by supervisors and trainees. Third, special therapeutic considerations will be provided to supervisors of therapists working with stigmatized populations. PMID- 10685357 TI - Milestones in periodontal research and the remaining critical issues. AB - A significant recent development in periodontal research has been the convergence of basic and clinical research resulting in a logarithmic increase in the rate of progress. Scientific consensus has been reached in many areas. In most populations, moderate to severe periodontitis affects a relatively small segment of adults who are at high risk. The microbial etiology is accepted and the identity of the major pathogenic bacterial species is known. The mechanisms through which resistant individuals successfully fend off the microbial challenge are known, and the immuno-inflammatory pathways activated by bacteria that underlie destruction of the alveolar bone and the connective tissues of the periodontium are reasonably well understood. The evidence shows that these pathways are held in common by all forms of periodontitis. Therapeutic modulation of these pathways is now possible, and new treatments based on such modulation are now becoming available. Although bacteria are essential for disease to occur, they are insufficient; a susceptible host is also necessary. Host susceptibility, disease progression and response to treatment are determined predominantly by heredity and environmental and acquired risk factors. Some of these can be changed while others are immutable. Concepts and procedures for treatment are generally scientifically based and appropriately applied. Preventive measures are largely successful and widely practiced in industrialized countries. Clearly, control of these ancient chronic diseases is now within our reach. In spite of the tremendous progress, many unresolved issues remain. The purpose of this paper is to summarize some of the major accomplishments of periodontal research, and identify and discuss some of the more important critical issues that still need to be addressed. PMID- 10685358 TI - Association of periodontal infections with atherosclerotic and pulmonary diseases. AB - Chronic infections may influence the severity and/or course of a number of systemic diseases. Periodontal diseases are localized chronic inflammatory conditions of the gingiva and underlying bone and connective tissues induced by bacteria and bacterial products of dental plaque. This paper will discuss the evidence for the role of periodontal disease in the pathogenesis of 2 important systemic diseases, atherosclerosis and pulmonary infections. Both epidemiological and laboratory studies are reviewed to assess the biological basis for the association of periodontal infections and these important diseases. Several potential mechanisms by which periodontal diseases may influence these conditions are also discussed. PMID- 10685359 TI - Periodontitis-atherosclerosis syndrome: an expanded model of pathogenesis. AB - The early reports of a linkage between periodontitis and atherosclerosis have garnered further support by additional data generated by several investigative teams in many different countries. The evidence continues to suggest that periodontitis may be an important risk factor or risk indicator for cardiovascular pathology for some individuals. The term periodontitis atherosclerosis syndrome (PAS) is proposed as a new diagnostic term to describe this condition in these individuals. Current evidence, albeit preliminary in nature, which describes a cluster of clinical signs and symptoms that are associated with this condition, is presented. It is clear that this syndrome will require considerable study and refinement before a definitive diagnosis and treatment plan can be formulated. Potential mechanisms by which systemic inflammation and infectious challenge of periodontal origin may serve as a potential modifier of cardiovascular disease are discussed in the context of a detailed working model of pathogenesis. This hypothetical model embraces many cellular and molecular components of atherogenesis and thromboembolic diseases from the perspective of periodontitis pathogenesis. Many aspects of the hypothetical model remain unproved; however, it is our opinion that only through the clarification of the mechanisms of pathogenesis can we ultimately construct a knowledge framework for accurate diagnoses and successful therapies. The concept of diagnosing and treating a periodontal patient to minimize the deleterious effects of this chronic infectious and inflammatory condition on the cardiovascular system represents an unprecedented challenge to our profession. PMID- 10685360 TI - Interleukin-1 genotypes and the association between periodontitis and cardiovascular disease. AB - An epidemiological association between periodontitis and cardiovascular disease has been reported in multiple studies. Various mechanisms have been proposed as potential explanations for this association, including a common factor that predisposes certain individuals to a hyper-responsive inflammatory response. Variations in the genes that regulate the interleukin-1 (IL-1) response have been associated with both periodontal disease and cardiovascular disease. New data indicate that one pattern of IL-1 genetic polymorphisms, characterized by the IL 1A (+4845) and IL-1B (+3954) markers, is associated with periodontitis but not certain measures of atherosclerosis. Another IL-1 genetic pattern, characterized by the IL-1B (-511) and IL-1RN (+2018) markers, is associated with atherosclerotic plaque formation, as measured by angiography and arterial wall thickness, but not periodontitis. These two patterns also have different functional implications relative to IL-1 biological activity. Studies of IL-1 gene polymorphisms, atherosclerotic plaque instability and cardiovascular clinical events are in progress. Hypothetical models are presented to explain how IL-1 genetic factors may be involved in cardiovascular disease. PMID- 10685361 TI - Systemic manifestations of periodontitis in the non-human primate. AB - This report describes our findings regarding the potential contribution of periodontitis to atherosclerotic processes using a nonhuman primate model. The goal of the investigations was to target general mechanisms which could describe the association of these disease processes, including: (i) systemic translocation of bacteria/products during periodontitis; (ii) alterations in systemic inflammatory biomarkers during periodontitis; and (iii) the relationship of periodontitis to serum lipids/lipoproteins. Increases in serum endotoxin (e.g. LPS) during ligature-induced periodontitis were observed in these animals. We determined serum levels of various acute phase reactants and chemokines (e.g. CRP, alpha 1-antitrypsin, haptoglobin, fibrinogen, IL-8). A number of these host factors were significantly increased during gingivitis and/or periodontitis. Finally, we observed specific changes in serum lipid levels (cholesterol, triglycerides, HDL, LDL) and lipoproteins (apoA-I) during periodontitis, which were exacerbated by exposure of the animals to a diet with elevated fat content. Thus, we have described systemic manifestations of periodontitis that include detection of bacterial products, inflammatory biomarkers, and dyslipoproteinemia consistent with an increased atherogenic risk. PMID- 10685362 TI - Potential mechanisms of susceptibility to periodontitis in tobacco smokers. AB - Tobacco smoking is probably the most important, controllable environmental risk factor in periodontitis. It results in changes in the vascular, inflammatory, immune and healing responses. The degree of exposure to tobacco smoking can be measured in pack years or by measuring serum cotinine and nicotine levels. In a previous paper we reported elevated levels of serum soluble intercellular adhesion molecule-1 (sICAM-1) in smokers, regardless of periodontal status. Elevated sICAM-1 has been found to be a risk marker for cardiovascular disease. In the present paper we report the short-term effects of an episode of smoking on blood flow and levels of sICAM-1. Human volunteers included non-smokers, light smokers and heavy smokers. Relative blood flow was monitored in the gingivae and forehead skin using a laser Doppler flowmeter and serum levels of sICAM-1, cotinine and nicotine measured before during and up to 60 min following an episode of smoking. We could not provide evidence to support the theory that there is localized vasoconstriction within the gingival tissues. In contrast, there was a significant increase in blood flow in the forehead skin of light smokers which was not observed in non-smoking controls or in heavy smokers, suggesting a long-term tolerance in this latter group. The level of sICAM-1 remained unchanged during this episode, further suggesting a long-term effect. In a parallel group of subjects, we were able to demonstrate a direct significant correlation between sICAM and serum cotinine levels. These observations may be relevant to aetiological mechanisms in periodontitis and other smoking-associated diseases. PMID- 10685363 TI - Lipoxin A4 and aspirin-triggered 15-epi-LXA4 inhibit tumor necrosis factor-alpha initiated neutrophil responses and trafficking: novel regulators of a cytokine chemokine axis relevant to periodontal diseases. AB - The impact of lipoxin A4 (LXA4) and aspirin-triggered-lipoxins (ATL) was investigated in tumor necrosis factor (TNF alpha)-initiated neutrophil (PMN) responses in vitro and in vivo using LX analogs that are metabolically more stable. At nanomolar levels, the LXA4 and ATL analog 15 R/S-methyl-LXA4 each blocked TNF alpha-stimulated IL-1 beta release by isolated human PMN in vitro. These LXA4-ATL actions were time- and concentration-dependent. The TNF alpha induced IL-1 beta gene expression was also regulated by 15 R/S-methyl-LXA4. In addition, 15 R/S-methyl-LXA4 added to murine air pouches dramatically inhibited TNF alpha-stimulated leukocyte trafficking in vivo, as well as altered the appearance of both macrophage inflammatory peptide-2 and IL-1 beta and concomitantly stimulated IL-4 in pouch exudates. These findings from in vitro and in vivo experiments indicate that both LXA4 and ATL are regulators of TNF alpha directed neutrophil actions and stimulate IL-4 in exudates and thus regulate mediators that are held to play an important role in the pathogenesis of periodontal disease. PMID- 10685364 TI - HLA genetics for diagnosis of susceptibility to early-onset periodontitis. AB - Human leukocyte antigens (HLA) are essential in the recognition of foreign antigens in humoral immune response, which is genetically predetermined. Susceptibility to certain diseases that involve the immune response has been studied in relation to distinct HLA types. Although some diseases have been found to correlate to specific HLA loci positively, it has been difficult to isolate HLA types that predispose patients to periodontal destruction. Here, we review the current knowledge and recent advances in HLA genetics and its biology, which determine susceptibility to early-onset periodontitis (EOP). The HLA-DRB1*1501 DQB1*0602 genotype has been found with increasing frequency in EOP patients. This HLA genotype expresses aspartic acid at position 57 and glycine at position 70 on the DQ beta chain, suggesting a capability to bind certain bacterial antigens. The T cell response against the outer membrane protein (Ag53) of Porphyromonas gingivalis was examined via this HLA genotype. Strong T cell response against Ag53 p141-161 was inhibited partially by anti-DR antibody, but not by anti-DQ antibody. Possible host and bacterial peptides capable of binding DRB1*1501 were elucidated when the peptide sequence was compared to gene and protein databases. These results suggest that patients who have the HLA-DRB1*1501-DQB1*0602 genotype may have an accelerated T cell response to certain periodontopathic bacteria such as P. gingivalis in hyperimmune reactions and thus increased susceptibility to EOP. PMID- 10685365 TI - Analysis of genetic polymorphisms at the interleukin-10 and tumour necrosis factor loci in early-onset periodontitis. AB - Early onset periodontitis (EOP) is considered to have a substantial genetic basis, although the gene or genes involved have not been elucidated. The aim of the present study was to investigate possible links between generalized EOP (GEOP) and genes regulating expression of the cytokines tumour necrosis factor (TNF) and interleukin-10 (IL-10). Microsatellite marker DNA sequences corresponding to phenotypic variations in cytokine response were analysed. Genotypic variations in cytokine response have been shown in vitro for TNF and IL 10, and specific alleles are implicated in diseases such as systemic lupus erythmatosus (SLE) and rheumatoid arthritis (RA). Two microsatellites at the IL 10 locus, IL10.R and IL10.G, and 1 microsatellite at the TNF locus, TNFa, were typed for 77 GEOP patients in the West of Scotland. Due to the highly polymorphic nature of the microsatellite loci, a statistical comparison with ethnically matched healthy controls (TNFa, n = 91, IL10.R, n = 94, IL10.G, n = 102) was conducted using a Monte Carlo simulation for each marker. No significant differences were observed for any of the 3 markers, although there were possible indications of trends similar to those observed in SLE for the IL10.G marker. In conclusion, no links were found between GEOP and microsatellites at TNFa, IL10.R or IL10.G loci. PMID- 10685366 TI - Staphylococcus aureus-induced inflammation and bone destruction in experimental models of septic arthritis. AB - Staphylococcus aureus is the most common cause of septic arthritis. This disease often leads to severe joint destruction and high mortality. An experimental model of S. aureus arthritis has been developed to study the course of inflammation and joint destruction, to elucidate the role of bacterial and host factors for joint pathology and mortality, and to develop therapeutical and preventive devices against septic arthritis and sepsis. Results show that the innate immune system is crucial in defending the host against staphylococcal infection while components of the specific immune system, T and B lymphocytes and their products, are detrimental to the host, mediating joint destruction and increasing mortality rates. Staphylococcal capsule polysaccharides, toxins, cell wall-attached adhesins and possibly also the chromosomal DNA are virulence determinants in S. aureus arthritis. Several vaccine candidates have recently been described which protects against staphylococcal infections, e.g. staphylococcal surface polysaccharides, enterotoxins devoid of their superantigenic properties and collagen adhesin. There are also new approaches suggested for treatment of ongoing infections, such as the combined use of antibiotics and corticosteroids. PMID- 10685367 TI - Porphyromonas gingivalis virulence factors and invasion of cells of the cardiovascular system. AB - Our laboratory is interested in the genes and gene products involved in the interactions between Porphyromonas gingivalis (Pg) and the host. These interactions may occur in either the periodontal tissues or other non-oral host tissues such as those of the cardiovascular system. We have previously reported the cloning of several genes encoding hemagglutinins, surface proteins that interact with the host tissues, and are investigating their roles in the disease process. Primary among these is HagA, a very large protein with multiple functional groups that have significant sequence homology to protease genes of this species. Preliminary evidence indicates that an avirulent Salmonella typhimurium strain containing hagA is virulent in mice. These data indicate that HagA may be a key virulence factor of Pg. Additionally, we are investigating the invasion of primary human coronary artery endothelial cells (HCAEC) by Pg because of the recent epidemiological studies indicating a correlation between periodontal disease (PD) and coronary heart disease (CHD). We found that some, but not all, strains of Pg are able to invade these cells. Scanning electron microsopy of the infected HCAEC demonstrated that the invading organisms initially attached to the host cell surface as aggregates and by a "pedestal" like structure. By transmission electronmicroscopy it could be seen that internalized bacteria were present within multimembranous compartments localized with rough endoplasmic reticulum. In addition, invasion of the HCAEC by Pg resulted in an increase in the degradation of long-lived cellular proteins. These data indicate that Pg are present within autophagosomes and may use components of the autophagic pathway as a means to survive intracellularly. However, Pg presence within autophagosomes in KB cells could not be observed or detected. It is therefore likely that Pg uses different invasive mechanisms for different host cells. This and the role of HagA in invasion is currently being investigated further. PMID- 10685368 TI - The rag locus of Porphyromonas gingivalis: a novel pathogenicity island. AB - Previous studies in our laboratories of the serum IgG antibody response of periodontal patients have demonstrated the presence of an immunodominant surface antigen (Mr 55 kDa) in the outer membrane of Porphyromonas gingivalis W50. Genetic analysis of this antigen revealed that the corresponding gene forms part of a small operon which may have arisen via horizontal gene transfer into the genome of this strain. On the basis of sequence homology, the 55 kDa antigen (RagB) and the product of a cotranscribed gene (RagA) may act in concert at the surface of the bacterium to facilitate active transport, mediated through the periplasmic spanning protein, TonB, or form part of a signal transduction system in this organism. The rag locus is present in only a proportion of P. gingivalis laboratory strains and clinical isolates. Analysis of the distribution of ragB in subgingival samples by PCR demonstrated that rag+ P. gingivalis are more frequently detected in deep periodontal pockets than shallow sites in periodontal patients. These findings indicate that the rag genes may influence the virulence potential of P. gingivalis strains which harbour this locus and may thus be considered a novel pathogenicity island. Furthermore, horizontal gene transfer between organisms in subgingival plaque may represent a significant force in the evolution of these bacteria with ramifications for both diagnosis and targeted treatment of periodontal disease. PMID- 10685369 TI - Evidence and a novel hypothesis for the role of dendritic cells and Porphyromonas gingivalis in adult periodontitis. AB - We have proposed a novel overall hypothesis and approach to understanding the pathophysiology of adult periodontitis, one of the most common diseases that afflicts the US population. While mortality of the dentition is the most familiar outcome of adult periodontitis, its links with other more severe diseases, including coronary artery disease, respiratory diseases and pre-term labor, cannot be ignored. We have called attention to the many intriguing parallels between adult periodontitis and contact hypersensitivity (CHS). CHS is among the most common of dermatoses that afflicts mankind and one of the most intensively studied of in vivo immune responses. Both adult periodontitis and CHS target the host integument (gingiva or skin) and appear to involve the activation and sensitization of similar subsets of antigen capture and presenting cells, the dendritic cells (DCs), as well as similar T cell subsets. DCs have been termed "nature's adjuvant", being more efficient at antigen-presentation than macrophages or B cells and the only antigen-presenting cells that can stimulate naive T cells to proliferate. This immunostimulatory capacity can also have detrimental effects for the host, as typified by graft-vs.-host disease and CHS responses. Both AP and CHS involve a predominantly destructive T cell response mediated by both regulatory and effector T cells. In the present paper, we show intriguing evidence that Porphyromonas gingivalis is a unique pathogen in this regard, able to infect, sensitize and activate DCs in vitro and, probably, in situ. Many questions about the role of P. gingivalis-sensitized DCs in adult periodontitis, and of the parallels between adult periodontitis and CHS, however, remain to be answered. PMID- 10685370 TI - Controlled delivery of inductive proteins, plasmid DNA and cells from tissue engineering matrices. AB - It has been estimated that half the annual health care budget in the United States is spent on patients suffering from tissue loss and late stage organ failure. Critical limitations inherent in traditional therapies call for novel tissue and organ replacement strategies. This paper discusses development of biomaterials for conductive, inductive and cell-based tissue replacement strategies. Biodegradable polymer scaffolds can be used as space-filling matrices for tissue development and barriers to migration of epithelial cells in tissue conductive approaches. Inductive approaches involve sustained delivery of bioactive factors, such as protein growth factors and DNA, to alter cell function in localized regions. Factors can be released from highly porous polymer scaffolds to allow factor delivery and tissue development to occur in concert. Cell-based approaches involve seeding of cells onto polymeric scaffolds in vitro and subsequent transplantation of the scaffold. New scaffold materials are being developed that address specific tissue engineering design requirements, and in some cases attempt to mimic natural extracellular matrices. These strategies together offer the possibility of predictably forming specific tissue structures, and may provide solutions to problems such as periodontal ligament detachment, alveolar bone resorption and furcation defects. PMID- 10685371 TI - Evolution of periodontal regeneration: from the roots' point of view. AB - Tissues lost as a consequence of periodontal diseases, i.e. bone, cementum and a functional periodontal ligament (PDL), can be restored to some degree. Nevertheless, results are often disappointing. There is a need to develop new paradigms for regenerating periodontal tissues that are based on an understanding of the cellular and molecular mechanisms regulating the development and regeneration of periodontal tissues. As one approach we have developed strategies for maintaining cementoblasts in culture by first determining the gene profile for these cells in situ. Next, cells were immortalized in vitro using SV 40 large T antigen (SV40 Tag) or by using mice containing transgenes enabling cellular immortality in vitro. Cementoblasts in vitro retained expression of genes associated with mineralized tissues, bone sialoprotein and osteocalcin, that were not linked with periodontal fibroblasts either in situ or in vitro. Further, cementoblasts promoted mineralization in vitro as measured by von Kossa and ex vivo using a severely compromised immunodeficient (SCID) mouse model. These cells responded to growth factors by eliciting changes in gene profile and mitogenesis and to osteotropic hormones by evoking changes in gene profile and ability to induce mineral nodule formation in vitro. The ultimate goal of these studies is to provide the knowledge base required for designing improved modalities for use in periodontal regenerative therapies. PMID- 10685372 TI - Regeneration of periodontal tissues by basic fibroblast growth factor. AB - Several growth factors (or cytokines) have recently received attention because of their ability to actively regulate various cellular functions of periodontal ligament (PDL) cells and the effects of topical application of such factor(s) on periodontal tissue regeneration has been evaluated. In this study, we examined the role of basic fibroblast growth factor (bFGF) in the wound healing and regeneration of periodontal tissues. Alveolar bone defects (such as 2-wall, 3 wall and furcation class II bone defects) were created surgically in beagle dogs and primates. Recombinant bFGF was topically applied to the artificial bony defects. Six or 8 wk after application, the periodontal regeneration was morphologically and histomorphometrically analyzed. In all sites where bFGF was applied, significant periodontal ligament formation with new cementum deposits and new bone formation was observed in amounts greater than in the control sites. We found it noteworthy that no instances of epithelial down growth, ankylosis or root resorption were observed in the bFGF sites. In vitro studies demonstrated that bFGF enhances the proliferative responses of human PDL cells, which express FGF receptor-1 and -2, but inhibits the induction of alkaline phosphatase activity and mineralized nodule formation by PDL cells. Interestingly, we observed that the mRNA level of laminin in PDL cells, which plays an important role in angiogenesis, was specifically upregulated by bFGF stimulation, but that of type I collagen was downregulated. The present study demonstrates that bFGF can be applied as one of the therapeutic modalities which actively induce periodontal tissue regeneration. The results of in vitro studies suggest that by suppressing the cytodifferentiation of PDL cells into mineralized tissue forming cells, bFGF may play important roles in wound healing by promoting angiogenesis and inducing the growth of immature PDL cells, and may in turn accelerate periodontal regeneration. PMID- 10685374 TI - Initial results from the European Network of Health Promoting Schools program on development of health education in Finland. AB - The European Network of Health Promoting School (ENHPS) program aims to foster healthy lifestyles for school populations by developing supportive teaching and learning environments conducive to promotion of health. Most European countries have joined the network since its organization in 1993. This paper describes how the European, national, and local aims of the ENHPS program have been realized in Finnish schools during the first year of the second triennium (1997-1999). Substantial development related to health promotion has occurred in the Finnish ENHPS schools. A safe school environment was emphasized, and networking with other schools was encouraged at the international and national levels. Attitudes toward the ENHPS program generally were positive. However, Finnish schools have emphasized developing "structures" for health promotion. In the future, efforts should concentrate on students' active participation in the activities of health promotion in everyday teaching and learning situations. PMID- 10685373 TI - Effect of prostaglandin E2 on recombinant human bone morphogenetic protein-2 stimulated osteoblastic differentiation in human periodontal ligament cells. AB - Recombinant human (rh) bone morphogenetic protein-2 (BMP-2) stimulates osteoblastic differentiation in cells isolated from human periodontal ligament (HPLC), and this action of rhBMP-2 may be modulated by prostaglandins (PGs), which are local regulatory factors in the bone metabolism. In the present study, we investigated the effect of prostaglandin E2 (PGE2) on rhBMP-2-stimulated osteoblastic differentiation in cultured HPLC. rhBMP-2 (500 ng/ml)-stimulated alkaline phosphatase (ALPase) activity was enhanced by simultaneous treatment with low concentrations (10(-10)-10(-8) M) of PGE2, whereas a high concentration (10(-6) M) of PGE2 suppressed it. rhBMP-2 did not induce cyclo-oxygenase-2 (COX 2) mRNA expression or subsequent PGE2 production, whereas it remarkably suppressed rhIL-1 beta-induced COX-2 mRNA expression and PGE2 production. The rhBMP-2 action on osteoblastic differentiation in HPLC was also enhanced by co treatment with 0.25 to 25 ng/ml of rh interleukin-1 beta (IL-1 beta). The ALPase activity stimulated by simultaneous treatment with rhBMP-2 and rhIL-1 beta was partially inhibited by addition of 10(-6) M of indomethacin, which completely inhibited rhIL-1 beta-induced PGE2 production. These results reveal that PGE2 at different concentrations exerts a biphasic effect on BMP-2-stimulated osteoblastic differentiation in HPLC, BMP-2 inhibits IL-1 beta-induced PGE2 production through suppressing COX-2 expression, and the BMP-2-stimulated osteoblastic differentiation may be enhanced by the endogenous PGE2 induced by BMP-2 and IL-1 beta. These suggest that BMP-2 action on osteoblastic differentiation in HPLC may be modulated by PGE2 in autocrine and paracrine fashions. PMID- 10685375 TI - Identifying the educational implications of chronic illness in school children. AB - Children and adolescents with chronic illness such as asthma, diabetes, and cancer are at the intersection of the health and education systems. At school, their health needs may be attended to by a school nurse, while their educational needs may be overlooked. This article helps school personnel become more proactive in identifying the educational implications of chronic illness in school children. The confusing array of terms used to refer to this population by health professionals and educators are clarified, and estimates of the size of the population are provided. The potential impact of some common disease processes, treatments, and medications are reviewed, as are the risks of chronic absenteeism. Suggestions for how school and health professionals can identify and work with this group of children are offered. PMID- 10685376 TI - Age of first use of cigarettes among rural and small town elementary school children in Illinois. AB - This study determined age of first use of cigarettes among rural and small town elementary school students. Data were collected from 1,950 elementary school students, grades kindergarten through sixth, attending seven different schools in southern Illinois. Bivariate odds ratios and multivariate logistic regression procedures identified risk factors of cigarette use among this elementary school population. A Duncan multiple-range test revealed no significant difference in cigarette use between grades kindergarten through fifth (average percentage of use for those grades was 4.7%), but use increased significantly to 17.4% in the sixth grade. Predictor variables with the greatest odds ratios were having tried alcohol (OR = 8), having tried chewing tobacco or snuff (OR = 4.4), and being in the sixth grade (OR = 2.2). Healthy People 2010 draft objectives emphasize prevention and reduction of tobacco use among youth. To be effective, tobacco prevention programs must begin in the elementary school years. PMID- 10685377 TI - Outcomes from a school-based nutrition education program alternating special resource teachers and classroom teachers. AB - This study modified a successful nutrition program to improve its transferability and potential for institutionalization. Specific aims were to determine: 1) if 16 nutrition lessons taught alternately by special resource teachers (SRT) and classroom teachers, could produce outcomes equivalent to 24 SRT lessons; and 2) teachers' reactions to the program. The quasi-experimental design used classrooms (19 treatment and 19 comparison) in matched schools. Surveys and plate waste measured children's outcomes, and classroom teachers were observed and interviewed. Treatment students showed greater knowledge and self-efficacy scores and consumed 0.36 more servings of fruits and vegetables at lunch. Behavioral differences between groups were greater when SRTs provided all instruction. Teachers supported the program and anticipated teaching more nutrition on their own, but noted serious structural barriers. Findings support the need for long term contact to induce behavior change and the advantage of using teachers specifically trained in nutrition and experiential education. PMID- 10685378 TI - Epidemiology of school injuries in Utah: a population-based study. PMID- 10685379 TI - Indications for a home standing program for individuals with spinal cord injury. AB - Additional analyses were conducted on a recently published survey of persons with spinal cord injury (SCI) who used standing mobility devices. Frequency and duration of standing were examined in relation to outcomes using chi square analyses. Respondents (n = 99) who stood 30 minutes or more per day had significantly improved quality of life, fewer bed sores, fewer bladder infections, improved bowel regularity, and improved ability to straighten their legs compared with those who stood less time. Compliance with regular home standing (at least once per week) was high (74%). The data also suggest that individuals with SCI could benefit from standing even if they were to begin several years after injury. The observation of patient benefits and high compliance rates suggest that mobile standing devices should be more strongly considered as a major intervention for relief from secondary medical complications and improvement in overall quality of life of individuals with SCI. PMID- 10685381 TI - Preliminary electrophysiological characterization of functionally vestigial muscles of the head: potential for command signaling. AB - In devastating neurological disorders, such as quadriplegia resulting from high level spinal cord injury, it is essential to focus on functions that have been spared and optimally exploit them to enhance the individual's quality of life. It follows that certain muscles, which prior to the paralysis of much of the rest of the body seemed to have no useful function, might be used to provide unique signals to control assistive devices. This report presents preliminary electrophysiological data demonstrating potentially useful myoelectrical signals from 3 functionally vestigial muscles in humans; the posterior, anterior, and superior auricular muscles. In phylogenetically lower species, these muscles serve to position the ear to enhance hearing. The auricular muscles receive their major innervation from cranial nerve VII and should not be compromised by even high-level spinal cord lesions. In this study, it was found that the muscles could be voluntarily activated and, by standard surface-electrode recording, had potentials ranging to 680 microV in amplitude. Posterior auricular muscle potentials were used to command a paddle in a computer ping-pong task that employed a CyberLink interface. The t values for accuracy scores and ball hits were both significant at the p = .0001 level. These facts indicate that the auricular muscles may be useful for controlling assistive devices. PMID- 10685380 TI - A comparison of FES with KAFO for providing ambulation and upright mobility in a child with a complete thoracic spinal cord injury. AB - This study compared functional and physiologic measures of ambulation and upright mobility with functional electrical stimulation (FES) versus knee-ankle-foot orthoses (KAFO) in an 11-year-old boy with a T-10 level spinal cord injury. The child was a limited community ambulator with bilateral KAFO and loftstrand crutches. The FES system consisted of percutaneous intramuscular electrodes controlled by a portable stimulator and thumbswitch, an AFO for ankle and foot support, and loftstrand crutches. The subject used a swing-through gait pattern with both modes of mobility. The Functional Independence Measure scoring system and time to completion were used to compare performance in 6 standardized activities: donning, high transfer, inaccessible toilet transfer, ascend/descend stairs, and floor-to-standing transfer. Ten repeated measures were performed for each mode. Physiologic measures included energy expenditure, postural stability using forceplates, and a Functional Standing Test (FST). The subject performed all 6 mobility activities independently with FES and KAFO. In 4 of 6 activities, there was a trend toward faster times with FES, but this was not statistically significant. Toilet transfers and stair descent were performed significantly faster with KAFO. There was no difference in completion times on the activities of the FST. Measures of postural sway suggested that the subject was more stable with KAFO during quiet standing, while the modes were equal during a dynamic activity (raising arm for functional use). Energy expenditure results revealed no significant difference in oxygen cost per meter but a significantly higher oxygen consumption rate per minute for FES. Ambulation with both modes was performed at levels consistent with strenuous exercise. Maximum ambulation distances were relatively equal while the subject's velocity was significantly faster with FES. Of note, the subject reported ceasing ambulation during maximum distance trials due to general fatigue when using FES and due to shoulder pain with KAFO ambulation. For this subject, FES provided a means of performing upright mobility tasks independently, comparable with that of KAFO, while providing a faster ambulation velocity and a potential means of cardiovascular training. PMID- 10685382 TI - Surgical treatment of posttraumatic cystic and tethered spinal cords. AB - Posttraumatic syringomyelia as a cause of progressive neurologic deterioration has been well described. More recently, the noncystic posttraumatic tethered cord has been associated with identical progressive neurologic deterioration. A retrospective analysis of patients treated surgically with spinal cord untethering and/or cyst shunting to arrest a progressive myelopathy from a posttraumatic tethered and/or cystic cord was performed. Emphasis was on outcome using the American Spinal Injury Association (ASIA) sensory and motor scoring systems. During an 18-month period from May 1993 to December 1994, 70 patients with spinal cord injury were operated upon for tethered and/or cystic spinal cords because of a progressive myelopathy and deteriorating ASIA sensory/motor scores. Fifty-nine patients had follow-up data 1 year postoperatively. At the 1 year follow-up, there was small improvement in light touch sensory scores (0.67 points), pinprick scores (1.3 points), and motor scores (0.41 points) demonstrating that the progression of the myelopathic process was arrested. Thirty-four of these 59 patients had no previous surgery to their spinal cords. At 1 year follow-up, light touch scores improved on average 2.38 points, pinprick scores 3.88 points (p < 0.05), and motor scores 1.47 points, suggesting better outcome with first-time surgery. Of this latter group, 64.3% regained a lost function, 62.5% saw improvement in spasticity, 55.6% had substantial improvement in neurogenic pain, and 95.8% felt that surgery prevented further neurologic deterioration. PMID- 10685383 TI - Prevention of venous thromboembolism in patients with spinal cord injury: effects of sequential pneumatic compression and heparin. AB - OBJECTIVE: To estimate the incidence of and risk factors for venous thromboembolism in patients with acute traumatic spinal cord injury (SCI) and evaluate the effectiveness of sequential pneumatic compression devices (SCD), gradient elastic stockings (GES), and heparin in preventing thromboembolism. DESIGN: Prentice's case-cohort design. SETTING: All patients admitted to our hospital between 1976 and 1995 with acute traumatic SCI. MAIN OUTCOME MEASURES: Demographic characteristics, venous thromboembolism risk factors, methods of surveillance and prophylaxis, and thromboembolic events during the first 6 weeks following injury. RESULTS: Venous thromboembolism occurred in 84 of 428 patients (19.6%). Venous thromboembolism increased from 21% between 1976 and 1979 to 31% between 1980 and 1984, then decreased to 16% between 1985 and 1989 and to 8% between 1990 and 1995. Routine surveillance for venous thromboembolism increased through 1983, and SCD/GES use increased after 1983, with a concurrent decline in incidence of thromboembolism. Multivariate analysis showed that SCD/GES reduced the risk of deep venous thrombosis (DVT) or pulmonary embolism (relative risk, 0.5; 95% CI, 0.28 to 0.90). Multivariate analysis suggested a reduced risk of DVT in patients receiving heparin therapy within the first 14 to 42 days after injury, but estimates of reduced risk were not statistically significant (p = .064 for first 14 days, p = .13 for heparin anytime). CONCLUSION: The SCD/GES combination and heparin are each effective in preventing venous thromboembolism in individuals' acute traumatic SCI. Effectiveness of heparin prophylaxis may be greatest during the first 14 days after injury, whereas benefit from SCD continues to 6 weeks after injury. PMID- 10685384 TI - Effect of oral ciprofloxacin on bacterial flora of perineum, urethra, and lower urinary tract in men with spinal cord injury. AB - A study was performed in 25 men with spinal cord injuries undergoing intermittent catheterization whose urine had > or = 10(5) bacterial colonies/ml to determine efficacy of ciprofloxacin in eradicating susceptible organisms from urine, urethra, and perineum. Cultures were obtained prior to, during, and 5 to 7 days after administration of 500 mg twice daily for 10 days. Organisms in urine were also present in the urethra and/or perineum in 20 cases. Susceptible bacteria disappeared from urine in all subjects; but at follow-up 12 had cultures positive for ciprofloxacin-resistant Gram-positive cocci, including 1 with methicillin resistant Staphylococcus aureus (MRSA), and 2 with ciprofloxacin-resistant Acinetobacter sp. Treatment significantly reduced Gram-negative bacilli in perinea and urethras, but ciprofloxacin-susceptible organisms were replaced by resistant staphylococci, including MRSA, enterococci, and Acinetobacter sp. We support use of ciprofloxacin for treatment of urinary tract infections in persons with spinal cord injury, but in view of supercolonization with resistant organisms, the drug should be reserved for symptomatic persons not likely to respond to other oral agents. PMID- 10685385 TI - Treatment alternatives for spinal cord injury related spasticity. AB - Spasticity is a disorder of the motor system that occurs after injury to the central nervous system, which may increase the disability of an individual with spinal cord injury (SCI). Treatment options detailed in this review include rehabilitation techniques and modalities, pharmacological options, injection techniques, intrathecal baclofen, and surgery. This review will hopefully help health professionals to be aware of the treatment alternatives available for patients with SCI related spasticity. PMID- 10685386 TI - Subspecialty certification in spinal cord injury medicine: past, present, and future. PMID- 10685387 TI - Ultrastructure of Pasteurella haemolytica-induced changes in gnotobiotic calf lung. AB - Gnotobiotic calves born and maintained in a germ-free environment until inoculated with a pathogen are model animals for studying the progression of a specific disease, such as pneumonic pasteurellosis. To investigate early progression of pneumonic pasteurellosis, we compared the ultrastructure of regions of gas-exchange in the lungs of three challenge-exposed and three uninoculated control gnotobiotic calves. Three calves were inoculated endobronchially with a bolus of 7.9 x 10(10) CFU of Pasteurella haemolytica A1 and studied in a specific pathogen-free environment until severe respiratory distress occurred, at which time they were euthanized. Slices of lung tissue from the midregion of the right dorsal caudal lobe (area of lesion) of infected and control calves were fixed in glutaraldehyde and prepared for scanning (SEM) and transmission (TEM) electron microscopy. SEM revealed bacteria among long tangled strands of fibrin in pulmonary alveoli that became obliterated with cellular debris. TEM revealed areas of encapsulated and/or nonencapsulated bacteria among the cellular debris and patches of fibrin. Many neutrophils and macrophages that infiltrated the alveoli had phagocytosed bacteria and undergone degradation. Less cellular damage was present when encapsulated bacteria bordered the interalveolar septa than when nonencapsulated lysed bacteria were present. Where lysed bacteria were present, the pulmonary capillaries were dilated because of swollen, degranulated neutrophils, fibrin clots, and cellular necrosis. Both encapsulated and nonencapsulated bacteria were present in the lung tissue of gnotobiotic calves within the first 24 h after endobronchial inoculation of early log phase P. haemolytica. Loss of capsular material around individual and divided pairs of bacteria led to their consequential aggregation, lysing, and severe damage to the adjacent pulmonary capillaries and interalveolar septa. PMID- 10685388 TI - Morphometric analysis of the endometrial epithelium of rats (Rattus norvegicus albinus) submitted to chronic experimental alcoholism. AB - The effects of alcohol ingestion on the epithelial layer of the uterine endometrium of rats submitted to experimental chronic alcoholism were observed by morphometric methods. Sixty adult rats (Rattus norvegicus albinus) of the same age (3 months) and weighing on average 228 g were divided into two groups. The control group received solid diet, rat chow, and tap water ad libitum. The experimental group received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30 degrees Gay Lussac (v/v). At the end of 90, 180 and 270 days of treatment, the animals at estro were anaesthetized with ethyl ether, weighed and sacrificed. The internal reproductive organs were dissected, weighed and fixed. The final mean body weights were similar in the control and alcoholic groups. The histological results showed intense atrophy of the lining epithelium of the endometrium of uterine horns in the alcoholic group. Morphometric analysis confirmed the endometrial epithelial atrophy, showing that the alcoholic group had small cytoplasmic and nuclear areas and small cytoplasmic and nuclear perimeter compared to the control group. Scanning electron microscopic images showed intense lipid droplets accumulation in the epithelial cells from alcoholic group. Therefore, we concluded that alcohol acts as a toxin on the epithelial layer of the rat endometrium. PMID- 10685389 TI - Effect of low temperature on mast cell exocytosis. Early cytoplasmic vesicle formation and the role of cytoskeleton. AB - Mast cells stimulated with adriamycin at 4 degrees C underwent a unique exocytotic reaction. Rat peritoneal cells including mast cells were stimulated in vitro with adriamycin (100 micrograms/ml) for 0, 10, 30 or 60 sec and observed by transmission electron microscopy. Early changes could be observed after 10 sec stimulation and consisted in an approximately 5-fold increase (p < 0.001) of 0.05 0.2 micron diameter cytoplasmic vesicles. The Golgi apparatus showed signs of activation and vacuolization. From 10 to 30 sec, cytoplasmic vesicles fused with the perigranular membranes and with the membranes of developing secretory channels. At 60 sec, the number of vesicles and vacuoles diminished to nearly two fold starting levels. The exocytotic reaction characteristically resulted in the formation of enormously dilated granular cavities. The secretory process appeared incomplete; after 60 sec, in fact, maximal histamine release was 20% and exocytosis could be found in approximately 30% of mast cells. Pre-incubation with vinblastine followed by adriamycin stimulation at 37 degrees C determined a dose dependent inhibition of histamine release which was accompanied by the ultrastructural appearance of numerous 0.05-0.5 micron cytoplasmic vesicles and by signs of inhibited exocytosis. Our results support the concept that hyperstability of the cortical cytoskeleton coupled with microtubule perturbation would be responsible for the depressed pattern of mast cell exocytosis observed at 4 degrees C. Although stimulation at 4 degrees C induces a paradoxal secretory process, we believe that this approach may represent a useful model for understanding some basic mechanisms of exocytosis in mast cells. PMID- 10685390 TI - Morphological features of regeneration of rabbit aortic endothelium after cryoinduced vascular damage. AB - Regeneration of endothelium after damage is an important factor, which limits the development of atherogenesis. This study examines the topographical characteristics of regenerating endothelial cells (EC) in rabbit aorta after de endothelialization by cryodestruction. The effects of cloricromene on these processes were also studied. Vessels were harvested from 6-month-old NZW rabbits, 1 and 3 days after cryodestruction. The vessels were evaluated using scanning electron microscopy (SEM). One day after cryodestruction, there were defects in the endothelial monolayer in the zone of injury in saline treated animals. Large numbers of platelets and monocytes were observed in association with endothelium in the damaged zone. Three days after de-endothelialization the size of the area of the damage had decreased. On the surface of the new endothelial layer and below this defect, the number of adhering monocytes was increased, and many microdefects between endothelium could be seen. Administration of cloricromene for 1 or 3 days after damage reduced the number of endothelium-adherent platelets and monocytes, and microdefects in endothelium. The feature of endothelial repair in rabbits is a relatively large involvement of monocytes and platelets, which are visible below regenerated endothelium. Administration of cloricromene essentially restored re-endothelialization and significantly decreased the number of adherent monocytes and microdefects in the new endothelium. PMID- 10685391 TI - Ultrastructural alterations in the adrenal gland cortex of mice experimentally infected with a Venezuelan isolate of Trypanosoma evansi. AB - The ultrastructural study of adrenal gland from mice experimentally infected with Trypanosoma evansi, in addition to intravascular and intracellular trypanosomes, showed different degrees of cortical cell alterations and capillary wall modifications. Beside its biological scope, these results suggest a role for the adrenal cortex to partake in Surra's etiopathogenesis and describe for the very first time a T. evansi intracellular stage. PMID- 10685392 TI - Lymphatic vessels of the human heart: precollectors and collecting vessels. A morpho-structural study. AB - Only topographic and distributional data are available on the lymphatic outflow vessels of the human heart. Here we describe their structural and ultrastructural features. Fragments of the atria, ventricles and fat surrounding the major coronary branches were obtained from hearts of dilated cardiomyopathy patients. Serial semithin sections were observed under light microscopy and used for tridimensional reconstructions. Ultrathin sections were observed by transmission electron microscopy. Precollectors, the initial lymphatic outflow routes of the heart, are small valved vessels with irregular, discontinuous musculature. They originate in the subepicardial region from a network of epicardial, and from scattered myocardial absorbing lymphatic vessels and drain into the collecting vessels accompanying the major coronary branches. Collecting vessels are larger but structurally similar to precollectors. Wall musculature is independent of the size of the vessel. Their ultrastructure is the same as that of precollectors. Endothelial cells have many Weibel-Palade bodies, cytoplasmic filaments and focal adhesions. The basement membrane is discontinuous and anchoring filaments are frequent and conspicuous. The subendothelial layer contains much elastin. Human heart collecting vessels and precollectors may only be distinguished by their size. The scarcity of musculature suggests that lymph progression in this district is mainly ensured by cardiac revolutions. Their ultrastructural features are determined by adaptation to dynamic forces. The architecture of these vessels (random, disorderly, discontinuous, lacking any exact plan) and their large variations in caliber are in line with the ontogenetic hypothesis that peripheral lymphatic vessels originate from the coalescence of mesenchymal lacunae. PMID- 10685393 TI - Ultrastructural identification of cells with dendritic cell appearance in atherosclerotic aorta of apolipoprotein E deficient mice. AB - The present electron microscopic study was undertaken to see whether cells with a dendritic cell appearance accumulate in atherosclerotic lesions of apolipoprotein E (apoE) deficient mice. Atherosclerotic aortas from 7 eight-month old apoE deficient mice were examined. In atherosclerotic plaques as well as in the underlying media and adventitia, cells with a dendritic cell appearance including the presence of a unique tubulovesicular system were detected. The tubulovesicular system was most hypertrophied in their cellular processes where the continuous cisterns sometimes formed circular structures. The cells containing the tubulovesicular system lacked lysosomes and phagolysosomes and their cytoplasm was free of lipid inclusions. The present observations suggest that dendritic cells are involved in apoE deficient mouse atherosclerosis. ApoE deficient mice might be a useful model for investigating functions of dendritic cells in atherogenesis. PMID- 10685394 TI - Cytotoxic effects of eosinophils on renal proximal tubular epithelial cells: implications for renal allograft rejection. AB - Eosinophils are inflammatory cells that actively participate in the defense against large, multicellular parasites. Eosinophils are also a prominent component in the inflammatory infiltrates seen in acute allograft rejection. Their role in that process, however, remains obscure. In this study we examined in vitro the direct cytotoxicity of eosinophils against renal proximal tubular epithelial cells (PTEC) in the presence or absence of antibody and complement. Eosinophils were able to attach to the PTEC surface in both circumstances, i.e., irrespective of opsonization, or other mediation by humoral factors. The release of eosinophil granule contents (including their characteristic crystalloids) on the target cell surface and resulting PTEC injury occurred also irrespective of the presence or absence of opsonization. Thus, based on these results, the potential of direct cytotoxicity of eosinophils for PTEC exists, particularly in situations where both of these cell types are 'activated'. PMID- 10685395 TI - Ultrastructural pathology in skeletal muscle of mice envenomed with Crotalus vegrandis venom. AB - In this ultrastructural study we examined skeletal muscle fibres from mice intraperitoneally inoculated with a sublethal dose of Crotalus vegrandis (rattlesnake) venom. The group of mice inoculated presented neurological symptoms characterised by respiratory failure and hind limbs paralysis. Skeletal muscle fibres showed different degrees of alterations. Most of them presented the characteristic pattern of necrosis in progress. Atrophied and hypercontracted fibres were frequently seen. Some atrophied and necrotic fibres showed several nucleoli-like bodies in the nucleus. In the atrophic and hypercontracted fibres, sarcoplasmic vacuolation and abnormal mitochondria with stacked cristae were observed. Areas of segmental necrosis were also frequently found. In connection with these altered muscle fibres, capillary abnormalities were detected. This study suggests that in envenomed mice respiratory failure symptoms may be related with muscle damage caused by Crotalus vegrandis venom components. PMID- 10685396 TI - A quick molecular method for the simultaneous detection in spermatozoa of nuclear, acrosomal and axonemal structure by fluorescent microscopy. AB - Ejaculated spermatozoa from infertile men presenting to our laboratory for semen analysis were processed with a new molecular method which reveals simultaneously, in the same sperm cell, the status of the acrosome, by testing the hyaluronidase content, the texture of the nucleus, by checking the DNA strands breaks, and the structure of the axoneme, revealing the tubulin content. The presence of hyaluronidase and tubulin is essential for the sperm function, and the analysis of the DNA status reveals the eventual apoptotic process. Using this method in normal spermatozoa, the staining of the acrosomal hyaluronidase reveals, by yellow-green fluorescence, the shape of the acrosomal complex and its texture. At the same time, in the same sperm cell, the staining of the axonemal tubulin demonstrates, by a red labeling, the presence of the protein and therefore the consistence of the axonemal structure. Simultaneously, at the head level, the absence of red labeling from nuclear DNA indicates that the apoptotic process is not present. This protocol allows quantification of the frequency of the presence of normal or abnormal spermatozoa, by an easy scoring and calculation of the apoptotic sperm or of the sperm with generic defects at acrosomal or flagellar level. The percentage of normal spermatozoa evaluated by the triple staining method has been compared with the results of the PAP staining and of the ultrastructural analysis, statistically elaborated. Triple staining results more severe than the PAP method, but TEM analysis is the finest technique to detect sperm abnormality because it considers the entire panel of sperm defects. PMID- 10685397 TI - Estrogen protects peripheral and cerebral blood vessels from toxicity of Alzheimer peptide amyloid-beta and inflammatory reaction. AB - Due to increases in life expectancy, women are living 30 years or more beyond menopause. This has led to an increasing interest in the association between postmenopausal estrogen deficiency and degenerative diseases associated with aging such as cardiovascular disease, osteoporosis and dementia. Women are two times more likely to develop late-onset Alzheimer's disease (AD) than age-matched men. A large number of observational reports and a few randomized clinical trials have indicated that estrogen replacement therapy (ERT) may retard the development and severity of dementia in postmenopausal women. The mechanism underlying the protective action of estrogen in AD is under active investigation. A chronic inflammatory reaction mediated by abnormal deposition of proteins such as amyloid beta (A beta) is central to the pathology of AD. We investigated the effect of low doses of conjugated estrogen (Premarin) in an animal model of A beta-induced vascular disruption and inflammatory reaction. This rodent model allows live videomicroscopic recording and electron microscopic analysis of peripheral vascular disruption and inflammatory reaction triggered by A beta. Estrogen prevented vascular deposition of A beta, endothelial and vessel wall disruption with plasma leakage, platelet and mast cell activation, and characteristic features of an inflammatory reaction: adhesion and transmigration of leukocytes. The beneficial effect was lost when estrogen treatment was discontinued. Estrogen also protected the cerebral blood vessels from endothelial dysfunction induced by A beta. This novel protective effect of estrogen against A beta cytotoxicity in peripheral and cerebral vasculature may contribute to the therapeutic efficacy of estrogen in AD and coronary vascular disease. PMID- 10685398 TI - Alteration of the follicular basement membrane in non-obese diabetic mice with spontaneous autoimmune thyroiditis. AB - The follicular basement membrane (FBM) prevents thyroglobulin from escaping to the peri-follicular space, where it can act as an antigen to induce experimental thyroiditis. Laminin, a component of the FBM, is responsible for directing cell migration and stimulates greater adhesion of activated T lymphocytes. Our purpose was to study the expression of laminin in the thyroid of NOD mice, which have a propensity for autoimmune diseases, including thyroiditis. Thirty NOD mice between 3 and 42 weeks old were studied. Eight had thyroiditis and 22 showed no inflammatory infiltration. An immunohistochemical examination using the streptavidin-biotin-peroxidase technique was conducted on paraffin-embedded tissue sections, with a polyclonal antilaminin antibody. Antigen retrieval was achieved through pepsin digestion and microwave irradiation in citrate buffer. Staining for laminin was restricted to the basement membrane. In thyroids with no infiltration, laminin was shown as a fine, continuous brown line in the basement membrane. In 6 out of the 8 cases of thyroiditis, clearcut interruption and destruction of the FBM was observed, particularly when the follicles were located in lymphocyte infiltrated areas or when there was fibrosis. There were significant alterations in the pattern of the FBM with extensive areas of discontinuity in the distribution of laminin. Such discontinuities could facilitate antigen exposure, especially thyroglobulin, which may contribute to autoimmune thyroiditis in NOD mice. PMID- 10685399 TI - Amorphous matrices in pathology: their definition and place in the spectrum of morphological variations in the vessel-stroma interface. (Re: Lugassy et al., 1998. Angio-tumoral laminin in murine tumors derived from human melanoma cell lines. Immunohistochemical and ultrastructural observations. J. Submicrosc. Cytol. Pathol., 30, 231-237) PMID- 10685400 TI - Construction of libraries for isolation of adrenergic receptor genes. PMID- 10685401 TI - Isolation of adrenergic receptor genes. PMID- 10685402 TI - Analyses of adrenergic receptor sequences. PMID- 10685403 TI - Polymerase chain reaction screening of genomic libraries for adrenergic receptor genes. PMID- 10685404 TI - Solution-phase library screening for identification of rare adrenergic receptor clones. PMID- 10685405 TI - Genetic polymorphisms of adrenergic receptors. PMID- 10685406 TI - Northern blot analyses detecting adrenergic receptor mRNAs. PMID- 10685407 TI - Ribonuclease protection assay for the detection of beta 1-adrenergic receptor RNA. PMID- 10685408 TI - Determination of adrenergic receptor mRNAs by quantitative reverse transcriptase polymerase chain reactions. PMID- 10685409 TI - Nuclear run-on assays for measurement of adrenergic receptor transcription rate. PMID- 10685410 TI - Primer extension methods for determination of beta 1-adrenergic receptor mRNA start sites. PMID- 10685411 TI - Use of eukaryotic vectors for the expression of adrenergic receptors. PMID- 10685412 TI - Expression of beta-adrenergic receptors in recombinant baculovirus-infected insect cells. PMID- 10685413 TI - Expression of beta-adrenergic receptors in E. coli. PMID- 10685414 TI - Use and pharmacological analysis of established and transfected cell lines expressing adrenergic receptors. PMID- 10685415 TI - Transient transfection and adrenergic receptor promoter analysis. PMID- 10685416 TI - Antisense RNA/DNA-based techniques to probe adrenergic receptor function. PMID- 10685417 TI - Targeted disruption of adrenergic receptor genes. PMID- 10685418 TI - Development of antibodies to adrenergic receptors. PMID- 10685419 TI - Western blot detection of adrenergic receptors. PMID- 10685420 TI - UV crosslinking of adrenergic receptors and ligands. Detection by SDS-PAGE. PMID- 10685421 TI - Detection of beta-adrenergic receptors by radioligand binding. PMID- 10685422 TI - Assessing adrenergic receptor conformation using chemically reactive fluorescent probes. PMID- 10685423 TI - Biochemical methods for detection and measurement of cyclic AMP and adenylyl cyclase activity. PMID- 10685424 TI - Assay of arachidonic acid release coupled to alpha 1- and alpha 2-adrenergic receptors. PMID- 10685425 TI - Patch-clamp recording methods for examining adrenergic regulation of potassium currents in ocular epithelial cells. PMID- 10685426 TI - Southwestern blots for detection of a DNA binding protein recognizing the alpha 1B-adrenergic receptor gene promoter. PMID- 10685427 TI - DNase I footprinting analysis of transcription factors recognizing adrenergic receptor gene promoter sequences. PMID- 10685428 TI - Electrophoretic mobility shift assay for detection of DNA binding proteins recognizing beta-adrenergic receptor gene sequences. PMID- 10685429 TI - Determination of mRNA stability and characterization of proteins interacting with adrenergic receptor mRNAs. PMID- 10685430 TI - Use of immunohistochemistry and confocal microscopy in the detection of adrenergic receptors. PMID- 10685431 TI - Distribution of alpha 2A- and alpha 2C-adrenergic receptor immunoreactivity in the central nervous system. PMID- 10685432 TI - Quantitative light microscopic autoradiography of alpha 2-adrenergic radioligand binding sites. PMID- 10685433 TI - In situ hybridization of adrenergic receptor mRNA in brain. PMID- 10685434 TI - Use of electron microscopy in the detection of adrenergic receptors. PMID- 10685435 TI - Family functioning and the adjustment of adolescent siblings in diverse types of families. PMID- 10685437 TI - Adolescent siblings in stepfamilies: functioning and adolescent adjustment. Marital satisfaction, relationships, and roles. AB - The current study is one of the few attempts to examine the quality of marital relations in long-remarried families in diverse types of stepfamilies and compare them to long-established marriages in never-divorced families using a multimeasure, multirespondent design, which included observations. Early in a remarriage the new stepparent is often in a position of greater marginality than the biological parent is, and the marital relationship may be less central than in nonstepfamilies. Thus, the important differences between first marriages and remarriages may be in the salience rather than in the quality of the marital relationship. This will be explored as relations among family subsystems and child adjustment are examined in subsequent chapters. PMID- 10685436 TI - Adolescent siblings in stepfamilies: family functioning and adolescent adjustment. PMID- 10685438 TI - Parent-adolescent relationships in nonstep-, simple step-, and complex stepfamilies. PMID- 10685439 TI - Sibling, half sibling, and stepsibling relationships in remarried families. PMID- 10685440 TI - Adolescent adjustment in nonstepfamilies and stepfamilies. PMID- 10685441 TI - Relations among relationships: a family systems perspective. PMID- 10685442 TI - Family functioning in nonstepfamilies and different kinds of stepfamilies: an integration. PMID- 10685443 TI - Stepfamilies: the intersection of culture, context, and biology. PMID- 10685444 TI - Microbiological characteristics and susceptibility patterns of strains of Rhodotorula isolated from clinical samples. AB - The members of the genus Rhodotorula show a marked ubiquity. In man, they have been isolated from faeces, nails, skin, sputum, digestive tract and adenoids, forming part of the normal human flora, although in recent years cases have been reported of both local and systemic infection by this yeast. There are virtually no studies in the literature on the sensitivity of this genus to the antifungal agents in common clinical use. Therefore, it is considered of interest to study the microbiological characteristics and the susceptibility patterns of Rhodotorula isolated from clinical samples. A total of 35 different strains of Rhodotorula were studied. In vitro susceptibility testing to 5-fluorocytosine, amphotericin B, ketoconazole, fluconazole and itraconazole was performed. All the strains were considered sensitive to 5-fluorocytosine, amphotericin B, ketoconazole and itraconazole and resistant to fluconazole. As a conclusion, we can state that all the antifungal agents tested, except fluconazole, are useful medicaments for the treatment of infections by the Rhodotorula genus. PMID- 10685445 TI - Purification of the endospores and sporangia of Rhinosporidium seeberi on Percoll columns. AB - Human rhinosporidial tissue was used as the source of the various developmental stages of Rhinosporidium seeberi--endospores with electron dense bodies, juvenile, and immature sporangia. After homogenisation in phosphate buffered saline (PBS) and removal of tissue fragments by centrifugation, the rhinosporidial bodies were isolated on centrifuged Percoll columns with gradients of densities or on triple-layered columns of varying density. The separated bands, after repeated washing in PBS gave bodies free from human tissue as shown on Leishman and PAS staining and indirect immunofluorescence with rabbit and human patients' anti-rhinosporidial sera. Sonicates of these bodies were tested on agarose gel for precipitation with antisera, and on SDS-PAG electrophoresis and Coomassie Blue staining. Percoll columns were shown to be capable of isolating these stages of R. seeberi, free from human tissue and contaminating bacteria. PMID- 10685446 TI - Female genital coccidioidomycosis (FGC), Addison's disease and sigmoid loop abscess due to Coccidioides immites; case report and review of literature on FGC. AB - We describe a woman with unusual complications of infection with Coccidioides immitis--infection of the genital tract and adrenal insufficiency. The patient also had intestinal coccidioidomycosis (cocci) in conjunction with presumed pulmonary, and asymptomatic central nervous system cocci. To our knowledge, concurrent FGC, intestinal and adrenal cocci have not been reported previously. A MEDLINE review from 1966-1997 revealed only 1 case of adrenal insufficiency due to cocci. FGC is rare; we identified 12 reported cases since 1929. No combination of investigations or clinical features is sensitive enough to predict FGC. Diagnosis is usually made after microscopy of surgical specimens. FGC presents either as tubo-ovarian disease or endometritis. Treatment generally involves surgical excision and antifungal agents. We hypothesize that an initial trial of antifungals may obviate the need for surgery. PMID- 10685447 TI - Endocarditis by Aspergillus fumigatus in a renal transplant. PMID- 10685448 TI - Comparative study of agar diffusion test and the NCCLS macrobroth method for in vitro susceptibility testing of Candida spp. AB - We performed a prospective double-blind study to evaluate the correlation between inhibition zones obtained by a disk-diffusion test, using Neo-sensitabs of fluconazole (Rosco Diagnostica), and the MICs generated by the NCCLS macrobroth dilution assay. Eighty clinical isolates, representing 5 of the clinically relevant species of Candida, were tested simultaneously by both methods. A clear inverse correlation was found between the results obtained by both tests (r = 0.69). In addition, there was high degree of agreement between methods in the identification of susceptible isolates. However, the resistance definition by disk-diffusion test had a positive predictive value of only 17%. Our data support the hypothesis that Rosco Fluconazole Neo-sensitabs have potential as a screening test for the identification of Candida isolates susceptible to fluconazole. Resistant isolates should be further investigated by standardized broth procedures. PMID- 10685449 TI - Prevalence of dermatophytoses in the Zarqa district of Jordan. AB - A total of 350 clinically suspected cases of dermatomycoses were examined for causative fungi during July 1997 to September 1998. Mycotic infection was demonstrable by microscopy and culture in 199 (56.8%) cases. The most common superficial mycotic infections were tinea pedis (35.2%) followed by tinea capitis (23.1%), tinea unguium (21.6%) and tinea corporis (10.6%). Most of the infected patients were 1-9, 20-29 and 30-39 years old. Men were mainly infected with tinea cruris and tinea pedis, while women were infected with tinea pedis, tinea unguium and tinea capitis. The frequencies of etiological agents isolated from patients were as follows: Trichophyton mentagrophytes var. interdigitale (32.7%), T. rubrum (28.6%), Epidermophyton floccosum (20.1%), Microsporum canis (11.1%), T. schoenleinii (4%), T. verrucosum (2%), T. violaceum (1%), and M. gypseum (0.5%). The number of infections varied with the seasons. The highest number of cases of tinea pedis and tinea cruris occurred in the summer months, while tinea capitis, tinea corporis and tinea unguium occurred in the spring and winter months. PMID- 10685450 TI - [Evidence-based medicine: from concept to practice]. PMID- 10685451 TI - [Analysis of hospitalization of adult sickle-cell patients in Guadeloupe]. AB - PURPOSE: To determine the characteristics of acute hospitalizations in adult patient with sickle-cell disease in Guadeloupe. METHODS: We retrospectively studied clinical features of adult patients followed up by the "Centre Caribeen de la Drepanocytose" (CCD) in 1996. Data were collected from the medical records of the hospitalized patients and the longitudinal records of the CCD. RESULTS: Sixty-three (25%) of the 251 patients who were followed up by the CCD required hospitalization in 87 cases (1.38 hospitalizations/patient). Mean age of the hospitalized patients was 27.5 years (range 17 to 71 years). Most hospitalizations involved men (29 [31%] vs 34 [22%] for women, P < 0.05), and most were for homozygous patients with sickle-cell anemia: 39 (31%) SS, 19 (18.55%) SC and five (21.75%) S beta thal. A painful vaso-occlusive crisis was noted in 67 episodes. There were nine acute chest syndromes (ACS), six of them occurred following a vaso-occlusive crisis. We noted 39 infectious episodes. The increase in C-reactive protein (> 100 mg/L) was associated with ACS or urinary infection. A patient with renal failure died during septicemia. CONCLUSION: This study confirms the need for prevention of painful crises and other severe complications in patients with sickle-cell disease. PMID- 10685452 TI - [Secondary cutaneous effects of hydroxyurea: prospective study of 26 patients from a dermatologic consultation]. AB - PURPOSE: Hydroxyurea is a treatment of myeloproliferative syndromes. Its cutaneous side-effects are underestimated, because they are usually benign. We undertook a prospective study to evaluate their frequency. METHODS: During a 2 year period, all patients taking hydroxyurea for more than 6 months who had consultations at the dermatology department were systematically examined, regarding cutaneous side effects. RESULTS: Twenty-six patients were examined. All but one had cutaneous side-effects, including dryness (n = 16), moderate alopecia (n = 2), increased skin pigmentation (n = 5), melanonychia, single (n = 1) or multiple (n = 7), cutaneous atrophy (n = 4), leg ulcers (n = 8), plantar keratoderma (n = 3), pseudodermatomyositis (n = 1), lichen planus-like eruption on the dorsum of the hands (n = 2), actinic keratosis (n = 8), squamous cell carcinomas (n = 2), and mouth ulcerations (n = 1). CONCLUSION: This study shows that the frequency of hydroxyurea cutaneous side-effects diagnosed in 95% of studied patients is underestimated. They are usually benign, but some of them, in particular leg ulcers and squamous cell carcinomas, lead to modification of the treatment (39% of studied patients). PMID- 10685453 TI - [Mechanisms of amyloidosis and the proteins involved]. AB - INTRODUCTION: Recent data in amyloid research have shed light on the amyloid substance and have broadened our knowledge on the mechanism of amyloid deposition. CURRENT KNOWLEDGE AND KEY POINTS: Despite uniform physical properties relating to the presence of beta-pleates, amyloid deposits are chemically heterogeneous and have different origins; additional types will probably be described in the future. Immunohistochemical techniques using specific antisera for each of the major protein present in fibrils could help greatly to subclassify these disorders. In most circumstances, a circulating precursor protein may result from overproduction of either intact or aberrant molecule, a reduction in its degradation or excretion, or genetic abnormalities associated with variant proteins. The cleavage of protein precursor molecules of the protein component of amyloid fibrils characterizes amyloidogenesis, though it is not necessary for some amyloidosis forms. This review summarizes advances in the understanding of the nature of amyloid substances, the mechanism of amyloid deposition and the principal pathogenic hypothesis. FUTURE PROSPECTS AND PROJECTS: SAP component is common in all amyloidosis and may be the target for future therapy. PMID- 10685454 TI - [Clinical forms of hepatitis A]. AB - INTRODUCTION: Although its incidence has decreased during the last 20 years, hepatitis A is still the most common hepatitis. In most cases, hepatitis A is asymptomatic. When it is symptomatic, the course is benign in most cases. The aim of this review is to summarize current data regarding unusual clinical forms of hepatitis A. CURRENT KNOWLEDGE AND KEY POINTS: Hepatic insufficiency is the most severe complication of hepatitis A. It is more commonly observed in adult patients. In most cases, the outcome of hepatic insufficiency is rapidly favorable. In rare cases, hepatic insufficiency progresses and encephalopathy subsequently occurs. At this stage, emergency liver transplantation may be necessary. Apart from hepatic insufficiency, the course of hepatitis A may be characterized by a relapse following initial improvement (relapsing hepatitis A) and prolonged cholestasis. FUTURE PROSPECTS AND PROJECTS: In regard to patients with liver insufficiency, physicians should be educated about the need to prevent all iatrogenic factors which could impair the outcome and to maintain a situation propitious to rapid liver regeneration, a necessary condition for recovery. When this prevention fails and liver transplantation has to be considered, auxiliary transplantation should always be considered because this procedure preserves the possibility of a delayed regeneration. The justification of systematic vaccination of patients with chronic hepatitis B, who could be at higher risk for hepatic insufficiency during hepatitis A, is controversial. PMID- 10685455 TI - [Non-surgical treatments of esophageal cancers]. AB - INTRODUCTION: Despite improvements in surgical techniques and perioperative mortality, only slight improvements in the 5-year survival of patients with esophageal cancer have been observed in the last 20 years. Many patients with apparently localized cancer will have recurrences or metastatic disease despite surgery with curative resection. Consequently, multimodal therapies, including chemotherapy and radiotherapy, were introduced. This review outlines and critically analyzes current non-surgical treatments, including palliative care. CURRENT KNOWLEDGE AND KEY POINTS: Esophageal cancers appear to be chemosensitive but the median duration of response is short and toxicity consistent, especially in metastatic disease. Consequently, palliative chemotherapy should be offered preferably within a clinical trial. Chemotherapy as the only adjuvant treatment cannot be recommended outside clinical trials. Radiotherapy alone as a curative treatment has been proven to be inferior to chemoradiotherapy in inoperable tumors. Some data support the use of preoperative chemoradiotherapy, but randomized trials are conflicting. A pathological complete response has been identified as a favorable prognostic factor for survival. Self-expanding esophageal metal stents are a simple and effective palliative treatment of malignant dysphagia and can be considered as the reference treatment in patients with obstruction of the lower esophagus or with fistula. FUTURE PROSPECTS AND PROJECTS: Taxanes should be evaluated in randomized studies using chemotherapy or chemo-radiotherapy. Progress in radiotherapy, such as accelerated fractionation, greater radiation dose, and the addition of brachytherapy, will increase locoregional control and probably survival. The role of secondary surgery in patients responding to chemoradiotherapy still needs to be answered. PMID- 10685456 TI - [Bronchiolitis obliterans with organized pneumonia: a rare complication of primary Gougerot-Sjogren syndrome]. AB - INTRODUCTION: Bronchiolitis obliterans organizing pneumonia (BOOP) is characterized by plugs of granulation tissue in bronchioles, alveolar ducts and alveoli. This pulmonary disorder has been reported in some cases in relation to drug consumption (D-penicillamine, amiodarone), with bacterial or viral infections (Mycoplasma pneumoniae, HIV), and with systemic diseases, such as rheumatoid arthritis. To our knowledge, only three cases of association BOOP Sjogren's syndrome have been reported. EXEGESIS: We report three new cases of BOOP. These patients presented a primary Sjogren's syndrome without clinical or biological abnormalities suggestive of other autoimmune diseases. Initial presentation was an acute pulmonary disorder mimicking a bacterial pneumonia. Two patients had cutaneous vasculitis and the third vasculitic neuropathy. Corticosteroid therapy was begun and was quickly successful. None of the patients presented a relapse of BOOP. CONCLUSION: The incidence of BOOP is probably underestimated in patients with primary Sjogren's syndrome without cutaneous vasculitis. In case of pneumonia with antibiotic resistance, an immunological mechanism should be considered. PMID- 10685457 TI - [Rheumatoid arthritis preceding microscopic polyangitis. Report of two cases]. AB - INTRODUCTION: Although joint manifestations are common in microscopic polyangiitis (MPA), including arthralgia reported in 15-65% of cases and arthritis in 6-17%, there have been only two published cases of polyarthritis as the first manifestation of the disease. We report on two new cases. EXEGESIS: A 71-year-old woman had symmetric polyarthritis of the hands which initially suggested the existence of seronegative rheumatoid arthritis. A 52-year-old woman had seropositive asymmetric oligoarthritis. The diagnosis was not established until renal insufficiency appeared, prompting a renal biopsy which showed in both cases an extra-capillary glomerulonephritis and an anti-myeloperoxydase (p-ANCA) assay which was postive in both patients. The incidence and specificity of antineutrophil cytoplasmic antibodies (ANCA), including MPA, in rheumatoid arthritis are reviewed. CONCLUSION: Our two observations show that in cases of polyarthritis or oligoarthritis with renal involvement, testing for and typing of ANCA should be performed so as not to misdiagnose vasculitis. PMID- 10685458 TI - [Cryptococcal meningitis associated with chronic lymphocytic leukemia: a case report. Review of the literature]. AB - INTRODUCTION: The authors report the occurrence of a cryptococcal meningitis in a patient treated by corticosteroids and polychemotherapy for a chronic lymphocytic leukemia. EXEGESIS: A 63-year-old man with chronic lymphocytic leukemia was sent to hospital because of impaired condition with fever. Neurological disorders appeared. Cryptococcal meningitis was diagnosed. Under treatment, the outcome was favorable. CONCLUSION: This paper highlights the feature of this infection most likely underestimated in HIV-seronegative patients and the need to a priori consider this diagnosis. PMID- 10685459 TI - [Hepatocellular carcinoma: a case revealed by metastatic orbital tumor]. AB - INTRODUCTION: We report a case of metastatic orbital tumor revealing hepatocellular carcinoma. EXEGESIS: Metastatic orbital tumors of hepatocarcinoma are rare. Only six cases have been reported. We compare these cases to our observation. Treatment of the orbital metastasis is important to decrease pain, ophthalmological symptoms and to improve the quality of survival. Radiotherapy and/or surgery can be used. Prognosis for life depends on liver involvement: the modalities of treatment of the hepatocarcinoma have to be discussed for each patient. CONCLUSION: Seven cases of orbital metastasis revealing a hepatocarcinoma have been documented. Effectiveness of radiotherapy makes the local prognosis good, but prognosis depends on liver involvement, since prognosis of hepatocellular carcinoma is poor. PMID- 10685460 TI - [Lung neoplasms revealing a rare cutaneous tumor: eccrine porocarcinoma]. AB - INTRODUCTION: Eccrine sweat gland carcinoma, which belongs to the eccrine sweat gland carcinoma family, is a rare malignancy of the skin with a potential aggressive growth and metastatic spread. EXEGESIS: We report here a case of malignant eccrine poroma arising on the upper leg, with widespread pulmonary metastases. CONCLUSION: A brief synopsis of the pathological and clinical aspects of eccrine sweat gland carcinoma is presented with the currently available therapies. PMID- 10685461 TI - [Olfactory disorders and general pathology. Analysis and review of the literature]. AB - INTRODUCTION: Disturbances of the sense of smell have been documented in many general pathologies. The actual etiology of such impairments is often difficult to determine. The aim of the authors is to review the literature on olfactory disorders in general diseases. CURRENT KNOWLEDGE AND KEY POINTS: Acute and chronic liver disorders are frequently associated with hyposmia, which can be improved by vitamin A intake. Renal insufficiency could induce hyposmia according to the severity of the renal disease. Olfactory disorders seem to regress after transplantation but not after dialysis. Patients with AIDS--especially neurological forms--often present with taste and smell impairments. Smell alteration can also be noted in hypothyroidism and pseudohypoparathyroidism. In addition, taste and smell impairments have been described in patients with adrenal insufficiency or Cushing's disease. Subsequent to glucocorticoid therapy, adrenal insufficiency can induce regressive olfactory hypersensitivity. Olfactory impairments in diabetic patients can be associated with diabetic macrovascular manifestations due to ischemic alterations in the olfactory neuroepithelium. Impairment of the sense of smell has been described in many systemic diseases such as Horton's arteritis and Sjogren's syndrome. FUTURE PROSPECTS AND PROJECTS: Olfactory disorders should be investigated in patients presenting one of the above-mentioned diseases. PMID- 10685462 TI - [From knowledge to clinical practice: the introduction of evidence-based medicine]. AB - INTRODUCTION: The paper presents the new paradigm for medical practice which has been emerging for a decade: evidence-based medicine. CURRENT KNOWLEDGE AND KEY POINTS: The paradigm proposes new skills for the physicians, de-emphasizing intuition and empiricism in medical practice, and proposes the intensive use of medical literature for the medical practice. FUTURE PROSPECTS AND PROJECTS: The article presents the main characteristics of the approach which is raising new important questions about the evolution of medical practice and the validity of the paradigm. PMID- 10685463 TI - [An acute ossification of the hip]. PMID- 10685464 TI - [Lymphatic duct compression by an endothoracic goiter: an exceptional observation]. PMID- 10685465 TI - [Polymyositis: a rare complication of interferon alpha therapy]. PMID- 10685466 TI - [Aortic Salmonella infection. A new observation]. PMID- 10685467 TI - [Mycobacterium avium-positive pulmonary sample and CD4+ lymphopenia not related to human HIV]. PMID- 10685468 TI - [Comparing two groups of patients]. AB - In medical research, comparing two groups of patients occurs frequently. The basic principles of comparison and the usual statistical tests--according to the types of variables and the choices of sampling methods--are presented. Elements needed for the computation of the size of a comparative study are given. PMID- 10685469 TI - [Candida pneumopathy: fact or fiction?]. PMID- 10685470 TI - [Patient knowledge of asthma: results of a national survey in pneumology]. AB - Knowledge of asthma and its treatment were evaluated in a survey of 1,000 asthmatics using a self-questionnaire with closed questions. As far as the disease was concerned, 87.4% of patients knew that the bronchi were the pathological organ and 70.6% that the disease persisted between attacks. Overall knowledge of the disease (pathological organ, persistence of the disease between attacks, chronic inflammation associated with acute bronchoconstriction) was adequate in only 25.6% of patients. It was associated with the educational level [higher > primary; p = 0.001], the grade of asthma [severe > mild; p = 0.007] and the number of medicines inhaled [(n > 3) > (N < 2); p = 0.001)]. As far as medicines were concerned, knowledge of their mechanisms of action varied according to therapeutic groups: antiallergics [78.6%], antibiotics [77.7%], antihistamines [74.5%], LA beta2-mimetics [45.9%], theophylline [31.9%], corticosteroids [31.7%], beta2-mimetics [24.8%]. No factor was statistically correlated with greater familiarity. However, only 7.2% of patients treated with metered-dose aerosols expressed handling problems and 78.8% of asthmatics questioned felt that they were sufficiently informed about their disease and its treatment. Information and education thus remain a priority in the management of asthma. PMID- 10685471 TI - [Serum procalcitonin and respiratory tract infections]. AB - The aim of our study was to evaluate the prognostic value of serum procalcitonine (PCT) assay in adult respiratory infections. Forty-nine patients admitted with pleurisy, community-acquired pneumonia, tuberculosis, infection were included in this prospective study. PCT was assayed on admission and discharge. Biological and clinical parameters of gravity were also evaluated. Twenty patients had elevated PCT of more than 0.50 ng/ml. In 29 patients, PCT was undetectable. The serum PCT level was normal in the patients with tuberculosis, infection, pneumocytosis. PCT did not correlate with the biological and clinical markers of the disease severity but the evolution of PCT correlated with the evolution of C reactive-protein (r = 0.58, p < 0.05). PCT seems to be an early marker of the evolution of respiratory infections, but it does not help to establish prognosis. Further studies are necessary to assess the potential value of PCT in more severe respiratory infections requiring assisted ventilation. PMID- 10685472 TI - [Diffuse interstitial pulmonary fibrosis and bronchial cancer]. AB - Mortality due to lung cancer was 25% (7/28) in this study of patients with diffuse interstitial pulmonary fibrosis. Opacities on the chest x-ray suggestive of lung cancer were observed in 5 of the 7 cases. All 7 had squamous cell carcinoma. The percentage of smokers was significantly higher in patients with pulmonary fibrosis who developed lung cancer than in those with fibrosis who did not develop lung cancer (p = 0.016). These 7 cases of lung cancer with pulmonary fibrosis were compared with 174 cases of lung cancer without associated fibrosis. Peripheral localizations and lower lobe involvement were higher in cases of lung cancer with pulmonary fibrosis. PMID- 10685473 TI - [Role of thoracic radiography in the management of community-acquired pneumonia]. AB - In patients presenting with a lower respiratory tract infection, it is generally recommended to order a chest X-ray in two circumstances: when the clinical signs suggest the diagnosis of pneumonia or in case of rather atypical symptoms suggesting a potential risk of complications. Indeed, the presence of one or more recent opacities, and more specifically, homogeneous alveolar infiltration(s) remains the gold standard criterion for the diagnosis of pneumonia. One must not however let this rule overshadow certain limitations of the chest X-ray reported in the literature: 1. the misdiagnosis of certain cases of pneumonia or bronchopneumonia seen early; 2. the extent of the infiltration gives only a very relative assessment of severity; 3. the characteristics of the infiltration are only relatively specific for etiological diagnosis. In patients with community acquired pneumonia diagnosed on the basis of clinical signs and radiographic findings, systematically performed series of follow-up X-rays have shown that the initial extension of the infiltration and the rate of its resolution contribute quite variably to the diagnosis of complications. The best indication for ordering a follow-up X-ray before 6 weeks remains an unfavorable clinical course. PMID- 10685474 TI - [Thyroid manifestations of sarcoidosis: a case report]. AB - Sarcoidosis is a systemic disease with many localizations. Thyroid involvement has been often described but rarely confirmed histologically. A common immune mechanism appears to be the cause. Thyroid sarcoidosis should be envisaged in patients with a thyroid nodule and mediastino-pulmonary involvement. We report the case of a 63-year-old woman with no past history who was hospitalized for dyspnea. Explorations evidenced a cold thyroid nodule associated with diffuse interstitial lung disease and mediastinal node enlargement. Pathology examination disclosed the sarcoid nature of the thyroid nodule and the lung lesions. PMID- 10685475 TI - [Hydatid thymic cyst. A case report]. AB - Hydatid thymic cyst is exceptional. The diagnosis is suspected by radiology and epidemiology. Serology tests provide variable results and surgery is the only treatment. We report a case of thymic hydatid cyst in a 20-year-old man who had no other localizations. Diagnosis was confirmed at surgery and by histology. PMID- 10685476 TI - [Bacteremia and Pasteurella multocida pneumonia revealing HIV infection]. AB - Pasteurella multocida is a major pathogen in local wound infections due to animal bites. Pneumonia and bacteremia are less frequent and usually associated with local or general impairment of host defenses. Scarce reports exist about Pasteurella multocida infections in HIV infected patients. We report here a case of hypoxemic pneumonia and bacteremia due to Pasteurella multocida revealing an HIV infection in a young man. PMID- 10685477 TI - Prognostic factors for time receiving workers' compensation benefits in a cohort of patients with low back pain. AB - STUDY DESIGN: Prospective inception cohort study. OBJECTIVE: To develop a prognostic model that predicts time receiving workers' compensation benefits for low back pain claimants. SUMMARY OF BACKGROUND DATA: As the cost and difficulty of managing low back pain escalate, any predictor of outcome is advantageous. METHODS: To obtain the outcome and predictor variables, patient data from two separate databases were linked: a clinical database and an administrative (Ontario workers' compensation) database. Claimants injured between January 1 and December 31, 1994, were included and observed for 1 year from the date of accident. The outcome variable was cumulative number of calendar days receiving benefits. RESULTS: Multivariable Cox proportional hazards regression (forward stepwise) showed eight significant predictors; five were associated with increased time receiving benefits compared with their reference groups: 1) working in the construction industry, 2) older age, 3) lag time from injury to treatment, 4) pain referred into the leg, and 5) three or more positive Waddell nonorganic signs. Three predictors were associated with reduced time receiving benefits: 1) higher values of questionnaire score, 2) intermittent pain, and 3) a previous episode of back pain. A predictive score was calculated to categorize claimants as at high or low risk for chronicity. When an arbitrary cutoff point was set at the 75th percentile of predictive score, negative predictive value was 94%. CONCLUSION: This research identified eight factors for time receiving workers' compensation benefits among claimants with low back pain. This model discriminates between high- and low-risk claimants. Few low-risk claimants continued to receive benefits for more than 3 months. PMID- 10685478 TI - Effect of augmentation on the mechanics of vertebral wedge fractures. AB - STUDY DESIGN: The effect of cement augmentation of wedge-fractured vertebral bodies on spine segment compliance was studied in 16 cadaver specimens. OBJECTIVES: 1) To assess the mechanical effects of cement augmentation of vertebral wedge fractures. 2) To determine whether a new reduction/injection procedure has the same mechanical effects as the established direct injection procedure. SUMMARY OF BACKGROUND DATA: Although wedge fractures cause pain and disability in hundreds of thousands of people, few effective treatments are available. Clinical studies have shown that cement augmentation, a new procedure, effectively relieves pain and restores mobility in patients suffering from weak or fractured vertebrae. However, only a few studies have examined the mechanics of vertebral augmentation. METHODS: A wedge fracture was created in the middle vertebra of 16 three-vertebra cadaver spine segments. Neutral and full-load compliance of each fractured spine segment in flexion/extension and lateral bending were assessed by measuring the relative rotation of the vertebral bodies in response to applied moments. Eight of the fractured vertebral bodies were then augmented using direct injection, while the remaining eight fractured vertebral bodies were augmented using a combined reduction/injection procedure. Compliance of the augmented segments was then assessed. RESULTS: Augmentation significantly reduced the neutral compliance (reduction of 25% +/- 23%) (mean +/- standard deviation) and the full-load compliance (reduction of 23% +/- 20%) in flexion/extension (P < 0.005). Augmentation also significantly reduced the neutral compliance (reduction of 34% +/- 20%) and the full-load compliance (reduction of 26% +/- 17%) in lateral bending (P < 0.0001). No significant difference was found between the two procedures for compliance reduction. CONCLUSIONS: Augmentation of wedge fractures using both direct injection and reduction/injection reduces spine segment compliance significantly. PMID- 10685479 TI - Changes with age in proteoglycan synthesis in cells cultured in vitro from the inner and outer rabbit annulus fibrosus. Responses to interleukin-1 and interleukin-1 receptor antagonist protein. AB - STUDY DESIGN: Proteoglycan synthesis was examined in cells isolated from the inner and outer annulus fibrosus of young and old rabbits. Their responses to interleukin-1 alpha and interleukin-1 receptor antagonist protein were investigated. OBJECTIVES: To evaluate the age-related changes and the anatomically related differences in the function of intervertebral disc cells. SUMMARY OF BACKGROUND DATA: Proteoglycan content in the human intervertebral disc decreases with age. Age-related changes in intervertebral disc cell function, however, have not been fully investigated. METHODS: Japanese white rabbits aged 2 months (young group) and 3 years (old group) were used. The inner and outer layer of the annulus fibrosus were separated. The proteoglycan synthesis and release were measured in cells cultured with or without human recombinant interleukin-1 alpha and interleukin-1 receptor antagonist protein. RESULTS: The proteoglycan synthesis significantly decreased and the release rate significantly increased in the old rabbits, compared with the young ones. In the inner annulus, the inhibition of proteoglycan synthesis due to interleukin-1 alpha was greater in the old rabbits than in the young ones. In the old rabbits, interleukin-1-induced inhibition was more pronounced in the inner annulus than in the outer annulus. Interleukin-1 receptor antagonist protein suppressed inhibition of proteoglycan synthesis by interleukin-1 alpha in the two layers in both age groups. CONCLUSIONS: Both the decline in proteoglycan synthesis and the increased cell sensitivity to interleukin-1 alpha with age may contribute to the degradation of discs. The increase in cell response to interleukin-1 alpha in the inner annulus of rabbits may explain why the inner annulus and nucleus pulposus degrade earlier than the outer annulus in human discs. Interleukin-1 receptor antagonist protein could be useful in inhibiting the degradation of the disc. PMID- 10685480 TI - Load-sharing characteristics of stabilized lumbar spine segments. AB - STUDY DESIGN: Load sharing in stabilized spinal segments was evaluated using sequential injury and stabilization with a posterior instrumentation system under an in vitro flexibility protocol. OBJECTIVE: To analyze the partitioning of applied loads between anatomic and implanted structures of lumbar functional spinal units stabilized with a posterior instrumentation system. To identify surgical indications for which the risk of fixator breakage in vivo is high. SUMMARY OF BACKGROUND DATA: Relatively few groups have experimentally measured the in vitro and in vivo forces and/or moments supported by posterior instrumentation systems, and no analysis, of the load sharing in these systems has been performed. This information will provide novel insight into implant fatigue life, and the degree to which the spinal anatomy is shielded from the applied load and will allow the verification of mathematical models for new injury scenarios. METHODS: Specimen kinematics were determined using an optoelectronic tracking system. Intradiscal pressure and the forces and moments supported by the implants were measured using, respectively, a needle-mounted pressure sensor and strain gauges mounted on the spinal implants. RESULTS: A large majority of the applied moments were supported by an equal and opposite force pair between the intervertebral disc and fixator rods in flexion and extension and an equal and opposite force pair between the left and right fixator rods in lateral bending. Torsional moments were shared approximately equally between the posterior elements, intervertebral disc, an equal and opposite shear force pair in the transverse plane between the right and left fixators and internal fixator moments. CONCLUSIONS: When posterior instrumentation devices are used to stabilize severe anterior column injuries, they are at risk of fracture secondary to reversed bending moments. PMID- 10685481 TI - Transverse-contour modeling of trunk muscle-distributed forces and spinal loads during lifting and twisting. AB - STUDY DESIGN: An electromyography-assisted biomechanical model was developed using electromyographic (surface and in-dwelling) data collected during asymmetric lifting and twisting activities. OBJECTIVES: To develop a biomechanical model of the lumbar region that considers the ability of the broad, flat muscles of the trunk (external obliques, internal obliques and latissimus dorsi) to activate different anatomic regions at different intensity levels and then uses this information to describe the spine reaction forces that result during lifting and twisting tasks. SUMMARY OF BACKGROUND DATA: Many biomechanical models of the lumbar region use single-vector representations for the external oblique, internal oblique, and latissimus dorsi muscles. This simplification limits the description of the complexity of the resultant forces produced by these muscles and does not consider their differential activation capacity. METHODS: Human subjects performed lifting and twisting exertions while muscle electromyographic activities were sampled at one location on the rectus abdominis and erector spinae muscles and at multiple locations on the latissimus dorsi, external oblique, and internal oblique muscles. These data were used in conjunction with in vivo digitized muscle origin and insertion points to predict muscle forces and spine loads through the use of the electromyography-assisted modeling method. RESULTS: The measures of model performance such as percentage of error (6-21%) in the prediction of the external torques, correlations (0.83-0.98) between internal and external torques and the values of predicted muscle force capacity were all similar to data collected in previous electromyography-assisted models, but the predictions of spinal loading, particularly shear forces, were quite different. The results have shown that by modeling the broad, flat muscles of the torso using multiple-force vectors, the calculated shear forces in the spine were reduced. CONCLUSIONS: The multivector, transverse-contour model developed in this research illustrates the importance of realistic multiple vector modeling and the importance of considering the selective-activation capacity of the abdominal oblique musculature. PMID- 10685482 TI - Clinicoradiologic study of cervical laminoplasty with posterolateral fusion or bone graft. AB - STUDY DESIGN: A retrospective study of cervical expansive laminoplasty for cervical myelopathy from a clinicoradiologic perspective. OBJECTIVE: To clarify the correlation among sagittal curvature of the cervical spine, cervical range of motion, sagittal plane translation, spinal cord atrophy, and myelopathic symptoms in patients who have undergone laminoplasty. SUMMARY OF BACKGROUND DATA: Laminoplasties were developed to diminish the undesirable effects of laminectomy, which include postoperative kyphotic changes and instability. However, the superiority of laminoplasty over laminectomy remains controversial. METHODS: Fifty-one patients with cervical spondylotic myelopathy or ossification of the cervical posterior longitudinal ligament who underwent laminoplasty were radiologically assessed before and after surgery. The index of the sagittal curvature, intervertebral range of motion, listhesis, and the transverse area of the spinal cord at the site of maximal compression were measured to evaluate interrelations among those parameters and myelopathic symptoms. RESULTS: There were no patients with kyphotic curvature before surgery. The postoperative curvature tended to be less lordotic. This tendency did not adversely affect postoperative symptoms. The intervertebral range of motion was significantly decreased except at C1-C2. The final C4-C5 range of motion and the postoperative myelopathic symptoms were negatively correlated. A significant correlation was observed between the postoperative spinal cord atrophy and the final myelopathic symptoms. CONCLUSIONS: The decrease in the lordotic curvature index and the decrease in the intervertebral range of motion after laminoplasty did not cause neurologic deterioration. In the C4-C5 intervertebral segment with a high incidence of listhesis, the restriction of the C4-C5 range of motion improved the clinical myelopathic symptoms. The radiologic prognostic factors were the postoperative restriction of intervertebral range of motion in preoperatively unstable segments and the anatomic reversibility of spinal cord insult. PMID- 10685483 TI - Nonoperative treatment for lumbar spinal stenosis. Clinical and outcome results and a 3-year survivorship analysis. AB - STUDY DESIGN: A cohort study of nonoperatively treated patients with lumbar spinal stenosis. OBJECTIVE: To assess the effectiveness of aggressive nonsurgical treatment for lumbar spinal stenosis. BACKGROUND DATA: While surgical treatment of lumbar spinal stenosis has been widely accepted, the natural history of this condition is poorly documented. Moreover, the effect of other available therapies is unclear. METHODS: Forty-nine patients meeting radiographic and clinical criteria for spinal stenosis underwent nonsurgical intervention consisting of therapeutic exercises, analgesics, and epidural steroid injections. Patients were followed for an average of 33 months. Outcome was assessed using a recently developed patient questionnaire for assessment of patients with lumbar spinal stenosis. Survival analysis was used to assess the probability of surgical intervention over the follow-up period. RESULTS: At 3 years following treatment, 9 of the 49 patients had undergone surgical intervention. Of the remaining 40 unoperated patients, it is reported that two suffered significant motor deterioration, one of whom still reported overall symptoms as mild improvement, and the other as definite worsening. Five of the 40 unoperated patients reported feeling overall symptoms as probably or definitely worse, 12 reported no change, 11 reported only mild improvement, and 12 reported sustained improvement. Twelve of the 40 unoperated patients also had none or only mild pain. CONCLUSIONS: The authors conclude that aggressive nonoperative treatment for spinal stenosis remains a reasonable option. PMID- 10685484 TI - A three-dimensional radiographic comparison of Cotrel-Dubousset and Colorado instrumentations for the correction of idiopathic scoliosis. AB - STUDY DESIGN: A prospective clinical study comparing two instrumentation systems for the correction of idiopathic scoliosis. OBJECTIVES: To measure the short-term three-dimensional changes in the shape of the spine after corrective surgery and compare the Cotrel-Dubousset instrumentation to the more recent Colorado instrumentation to determine whether one system provides better three-dimensional correction. SUMMARY OF BACKGROUND DATA: Adequate three-dimensional correction of scoliotic deformities has been reported with the Cortrel-Dubousset instrumentation system. During the past decade, a new generation of more versatile and user-friendly spinal implants has appeared, but there are no reports available to indicate whether similar or better correction can be obtained with these newer systems. METHODS: The three-dimensional geometry of the thoracic and lumbar spine was documented in the standing position using a three dimensional reconstruction technique based on multiplanar radiography in 67 adolescents with idiopathic scoliosis undergoing correction by a posterior approach. Changes in spinal shape were measured 3 days before and 1 month after the surgery in 31 patients with Cotrel-Dubousset instrumentation and 36 patients with Colorado instrumentation. RESULTS: In both groups, adequate three dimensional correction of the scoliotic deformities was documented for thoracic and lumbar curves, with significant changes in the frontal plane, in the plane of maximum curvature, and in its orientation. When comparing both groups, better correction was obtained in the frontal plane with the Colorado instrumentation (65% vs. 48% with Cotrel-Dubousset), a finding that may be explained by the significantly greater proportion of pedicle screws used in this group. CONCLUSION: Both instrumentation techniques achieve an effective and comparable three-dimensional correction of the scoliotic deformities. PMID- 10685485 TI - The relationship between tight hamstrings and lumbar hypolordosis in children with cerebral palsy. AB - STUDY DESIGN: Retrospective clinical and radiographic review. OBJECTIVE: To assess the influence of tight hamstrings on the sagittal alignment of the thoracic and lumbar spine in children with cerebral palsy. SUMMARY OF BACKGROUND DATA: It is postulated that tight hamstrings may produce a hypolordosis of the lumbar spine. The abnormal sagittal contour of the spine may lead to increased stresses in the lumbar spine and subsequent pain and disability. This is of special concern in children with cerebral palsy who often have shortened spastic hamstring muscles. METHODS: Twenty-one patients were evaluated, with a mean age of 9.4 years. Standing and sitting lateral spine films were obtained and the lumbar lordosis and thoracic kyphosis were measured using the Cobb method. The popliteal angle was measured to assess hamstring tightness, such that a large popliteal angle indicates tight hamstrings. RESULTS: We found a statistically significant correlation between the sitting lumbar curve and popliteal angle (Pearson correlation value -0.77, P < 0.01). As the popliteal angle increased, the amount of lumbar lordosis decreased. This correlation was less significant when the patient was standing (Pearson correlation value -0.59). CONCLUSION: This study demonstrates that there is a correlation between tight hamstrings, as measured by the popliteal angle, and decreasing lumbar lordosis, especially when sitting. PMID- 10685486 TI - A radiostereometric analysis of the movements of the sacroiliac joints in the reciprocal straddle position. AB - STUDY DESIGN: A Radiostereometric analysis of the reciprocal straddle position. OBJECTIVES: To evaluate the magnitude of rotation in the sacroiliac joints in the reciprocal straddle position. SUMMARY OF BACKGROUND DATA: The reciprocal straddle position has been objectified in different studies, using different techniques, to show a sacroiliac motion between 5 degrees and 36 degrees. Previous studies with radiostereometric analysis during different provocations reported much smaller movements. METHODS: Six women with posterior pelvic pain of long duration after pregnancy (n = 5) and sacroiliitis (n = 1) underwent radiostereometric analysis in the sustained reciprocal straddle position. RESULTS: A reciprocal movement could be demonstrated in the sacroiliac joints in the reciprocal straddle position. However, the movements were 10 times smaller than reported in earlier studies of the reciprocal straddle position. CONCLUSIONS: It was possible to demonstrate reciprocal movements of the sacroiliac joints in the straddle position. However, the radiostereometric analysis technique showed the movements to be small, as reported in other mobility studies. PMID- 10685487 TI - A cross-sectional study correlating lumbar spine degeneration with disability and pain. AB - STUDY DESIGN: Cross-sectional design. OBJECTIVES: To investigate the correlation between degeneration in the lumbar spine and self-reported disability and pain levels in patients with and without a history of trauma. SUMMARY OF BACKGROUND DATA: The link between lumbar spine degeneration and low back pain remains controversial, as does the correlation between trauma and spinal degeneration. METHODS: Radiographic and questionnaire data were collected from 172 consecutive patients with low back pain. Back pain severity was measured using two scales: one for pain over the entire episode and one for pain during the previous week. All patients also completed the Revised Oswestry Disability Questionnaire before radiography was performed. Further questions concerning the chronicity of symptoms and trauma were included. RESULTS: Controlling for age, patients with low back pain with a history of trauma had a statistically significant increase in the severity of facet degeneration (P < 0.02) compared with nontrauma patients with low back pain. However, there was no difference in disability and pain scores between the trauma and nontrauma patients or between the genders. A weak correlation between pain severity ratings and the number of levels of degeneration and the severity of the degeneration at the disc and facets was noted. CONCLUSIONS: Patients with low back pain with a history of trauma had more severe facet arthrosis than do nontrauma patients with low back pain, but there were no differences in pain and disability. There was a weak correlation between the quantity and severity of lumbar degeneration with pain levels, but not with disability scores. These findings are discussed in the light of recent reports regarding the cervical spine. PMID- 10685488 TI - Body weight and low back pain. A systematic literature review of 56 journal articles reporting on 65 epidemiologic studies. AB - STUDY DESIGN: A systematic review of the epidemiologic literature. OBJECTIVE: To establish if body weight is truly associated with low back pain (LBP) and whether the link may be causal. SUMMARY OF BACKGROUND DATA: Because obesity and LBP are prevalent in western society, it is of interest to establish whether obesity can induce LBP. METHODS: Fifty-six original research reports, reporting on 65 studies published between 1965 and 1997, were systematically reviewed for the frequency of positive associations between body weight and LBP. In addition, the presence of positive findings was examined in relation to several study characteristics. Based on these results, only studies emanating from the general population with a sample size exceeding 3000 were included in the additional search for causality using some of the classical Bradford-Hill criteria. The review was carried out by the author, blindly at 2 months' interval. RESULTS: Thirty-two percent of all the studies report a statistically significant positive weak association between body weight and LBP. Studies that fulfilled the post hoc criteria never report a rate ratio above 2, but there is possibly a positive biological gradient. These studies had no information on temporality or reversibility and there was no obvious consistency of findings. CONCLUSIONS: Due to lack of evidence, body weight should be considered a possible weak risk indicator, but there is insufficient data to assess if it is a true cause of LBP. PMID- 10685489 TI - Results of a multimodal treatment program for patients with chronic symptoms after a whiplash injury of the neck. AB - STUDY DESIGN: A descriptive case series pre- and post-treatment design, including a 6-month follow-up. OBJECTIVES: The objective of this study was to document the improvements of patients with chronic symptoms after a "whiplash" injury of the neck, who attended a 4-week multimodal treatment program at the Rug AdviesCentra Nederland. SUMMARY OF BACKGROUND DATA: To the authors' knowledge, no studies have been conducted on the effectiveness of multidisciplinary treatment of chronic symptoms after whiplash injury. METHODS: Twenty-six patients who experienced Quebec type 1 or 2 lesions of the neck (whiplash) with persisting symptoms of longer than 6 months' duration participated in the study. The measures included were pain intensity (according to the visual analog scale), number of painful sites (determined by pain drawing), self-reported disability Quebec Back Pain Disability Scale; and symptoms of somatic and psychological distress and cognitive symptoms (according to selected Minnesota Multiphasic Personality Inventory-2 scales). Furthermore, objective outcome criteria were used regarding return to work, medication, and medical and paramedical treatment. Statistical and clinical significance of treatment results were both assessed. RESULTS: The patients' symptoms improved significantly on nearly all self-report measures. Their scores for objective outcome criteria reported during the 6-month follow-up evaluation were: complete return to work (65%); complete or partial return to work (92%); no use of analgesics in the past 6 months (58%); and no medical or paramedical treatments in the past 6 months (81%). CONCLUSION: These early results indicate that a multimodal treatment program has the potential to be an effective treatment for patients with chronic symptoms after a whiplash injury of the neck--a group of patients who have in the past been considered intractable or, at the very least, puzzling. PMID- 10685490 TI - Repositioning error in low back pain. Comparing trunk repositioning error in subjects with chronic low back pain and control subjects. AB - STUDY DESIGN: Repositioning error of the trunk was tested in 20 subjects with chronic low back pain and in 20 control subjects. The 3Space Tracker (Polhemus, Colchester, VT), a device that measures three-dimensional position in space, was used to determine the subject's trunk position. OBJECTIVES: To determine whether repositioning error is different in subjects with chronic low back pain than in control subjects. SUMMARY OF BACKGROUND DATA: Proprioception allows the body to maintain proper orientation during static and dynamic activities. In peripheral joint injuries, researchers have demonstrated a loss of some aspects of proprioception and improvement in outcome with retraining. Although the components of proprioception in subjects with low back pain have not been well studied, it is thought that these persons lose some elements of proprioception that can be measured in a quantifiable way. If so, then rehabilitation to improve these deficits is important. In this pilot study, one aspect of proprioception, repositioning error, was examined. METHODS: The subjects attempted to replicate target positions of the trunk in flexion, extension, lateral bending, and lateral rotation. Repositioning error was calculated as the absolute difference between the actual and the subject-replicated target positions. RESULTS: No significant difference was found in repositioning error between the control subjects and the persons with chronic low back pain. CONCLUSIONS: Because proprioception is complex and entails the use of many afferent receptors, it is difficult to measure any one afferent deficiency discretely. The authors believe that this study, in which one aspect of proprioception was measured in an indirect manner, provides important background information on low back position sense. Further studies analyzing aspects of proprioception in subjects with low back pain are recommended. PMID- 10685491 TI - Coordination of primary health care for back pain. A randomized controlled trial. AB - STUDY DESIGN: A randomized controlled trial comparing usual care with a program for the coordination of primary health care (CORE) for the treatment of subacute low-back pain patients. OBJECTIVES: To measure the effectiveness of the CORE program as a mean for implementing clinical practice guidelines for low-back pain in an urban community. SUMMARY OF BACKGROUND DATA: Clinical practice guidelines have been developed for primary care physicians and patients on the clinical management of low-back pain. The implementation of the guidelines in a large community is difficult with the multiplicity of medical and nonmedical back care providers and products. The CORE program was designed to make the guidelines fit in this complex environment. METHODS: One hundred ten workers compensated for low back pain for 4 to 8 weeks in metropolitan Montreal were randomized in two groups: usual care (N = 56) and the CORE program (N = 54). Coordination of primary health care was performed by two primary care physicians and a nurse in liaison with the treating physicians, and included a complete examination, recommendations for the clinical management, and support to carry out the recommendations. All workers were followed for 6 months. Back pain and functional status were assessed at baseline, 3 months, and 6 months. RESULTS: In the 6-month follow-up, the CORE group returned to work 6.6 days (standard error = 8.9) quicker than the control group, a difference that was not statistically significant. However, the CORE group showed a sustained improvement in pain and functional status with two-fold differences at the end of the 6 months of follow up. This represented nine points on the Oswestry scale (P = 0.02) and 12 points on the Quebec Back Pain Disability Scale (P = 0.01). The CORE group also used three times less specialized imaging tests of the spine at 3 months (P < 0.01) and exercised twice as much at 6 months (P < 0.05) than the controls. CONCLUSIONS: The therapeutic results for workers with low-back pain could be improved by implementing the clinical practice guidelines with primary care physicians in a large community, without delaying the return to work. The CORE intervention for back pain patients is highly relevant to primary care practice. It is simple in its application, flexible to accommodate physicians' and- patients' preferences in health care, and it is effective on patients' clinical outcome. PMID- 10685492 TI - Endoscopic foraminotomy using MED system in cadaveric specimens. AB - STUDY DESIGN: Four cadavers had cervical foraminotomies performed at noncontiguous levels using either the standard open technique or the microendoscopic technique. OBJECTIVES: To evaluate the feasibility of using a minimally invasive technique for posterior decompression of cervical disc disease. SUMMARY OF BACKGROUND DATA: Even though the anterior approach is more commonly performed for the treatment of cervical disc disease, the posterior approach has distinct advantages in selected cases of foraminal stenosis and posterolateral disc herniation. Current technique, however, requires extensive muscle dissection, and is, therefore, subject to significant morbidity. METHODS: Each of four cadavers had posterior cervical foraminotomies performed using either the MICROENDOSCOPIC (MED) technique, or the standard open technique. Three noncontiguous levels were decompressed using one technique, and the other technique was used for the adjacent contralateral decompression. Each specimen was then evaluated with postoperative myelogram/CT and open dissection. Laminotomy size, length of root decompressed, and percentage of facet removed were measured. RESULTS: Average vertical diameter decompression and percentage of facet removed were significantly greater for the MED technique than for the open technique. Transverse diameter of the laminotomy area and the average length of decompressed root were not significantly different between the techniques. CONCLUSION: Posterior cervical foraminotomy, using the microendoscopic technique, is technically feasible and may be applicable to the treatment of foraminal stenosis and laterally located cervical disc herniation. Studies in live animals are currently examining techniques for hemostasis. PMID- 10685493 TI - Posterior spinal instrumentation and fusion of a neuromuscular scoliosis in a patient with autosomal dominant osteopetrosis. AB - STUDY DESIGN: A case report of a patient with autosomal dominant osteopetrosis and neuromuscular scoliosis who required surgical instrumentation and fusion of her spine. OBJECTIVE: To illustrate the surgical technique and long-term outcome in this rare form of spinal deformity. SUMMARY OF BACKGROUND DATA: Osteopetrosis is a group of rare skeletal dysplasias characterized clinically by skeletal osteosclerosis that is classically described in appearance as "marble bone." Despite the ubiquitous involvement of the vertebra, clinical manifestations of spinal involvement are uncommon. We present the case of an osteopetrotic patient with neuromuscular scoliosis who required surgical correction of her progressive deformity. There are no prior reports in the literature concerning operative or nonoperative management of scoliosis in this patient population. METHODS: The surgical technique utilized as well as the patient's response to surgical management of her scoliosis is presented with 5 year follow-up. RESULTS: The patient underwent a successful T4 to L1 posterior spine fusion and instrumentation using Luque rods, sublaminar wires and allograft bone augmentation. At 5 years following her index procedure, she is clinically and radiographically fused. CONCLUSION: Patients with osteopetrosis present unique surgical challenges during surgical correction of spinal deformities. The use of segmental sublaminar wires with 1/4-inch rods and crosslinks afforded stable fixation despite poor bone quality. Allograft bone combined with postoperative bracing resulted in a well-maintained correction and a solid fusion. Five year follow-up and continued radiographic evidence of stable fusion indicate that the presented approach can lead to a successful outcome in the osteopetrotic patient population. PMID- 10685494 TI - Giant cauda equina schwannoma. A case report. AB - STUDY DESIGN: Case report. OBJECTIVES: To present a rare case of a giant schwannoma of the cauda equina. SUMMARY OF BACKGROUND DATA: Giant spinal schwannoma of the cauda equina, which involves many nerve roots, is rare and there is usually no ossification in the schwannoma. It is unknown whether or not complete excision is preferable if the tumor is located in the lumbar lesion. METHODS: A 57-year-old woman had a 10-year history of low back pain. Scalloping of the posterior surface of the vertebral bodies from L3 to the sacrum was found. Magnetic resonance imaging disclosed a giant cauda equina tumor with multiple cysts. Central ossification revealed by computed tomography and an unusual myelogram made the preoperative diagnosis difficult. RESULTS: The patient underwent incomplete removal of the tumor, decompression of cysts, and spinal reconstruction. The tumor was proved to be a schwannoma. The postoperative course was uneventful and she has been almost free from low back pain for 3 years and 4 months. CONCLUSIONS: Giant schwannoma in the lumbar spine region is usually excised incompletely, because complete removal had the risk of sacrificing many nerve roots. In spite of the incomplete removal of the tumor, the risk of recurrence is low. PMID- 10685495 TI - Persistent osteopenia in adolescent idiopathic scoliosis: a longitudinal follow up study. PMID- 10685496 TI - Re: The graf ligamentoplasty procedure. Comparison with posterolateral fusion in the management of low-back pain. PMID- 10685497 TI - Synthesis of 6-aryloxy- and 6-arylalkoxy-2-chloropurines and their interactions with purine nucleoside phosphorylase from Escherichia coli. AB - The phase transfer method was applied to perform the nucleophilic substitution of 2,6-dichloropurines by modified arylalkyl alcohol or phenols. Since under these conditions only the 6-halogen is exchanged, this method gives 2-chloro-6-aryloxy- and 2-chloro-6-arylalkoxy-purines. 2-Chloro-6-benzylthiopurine was synthesized by alkylation of 2-chloro-6-thiopurine with benzyl bromide. The stereoisomers of 2 chloro-6-(1-phenyl-1-ethoxy)purine were obtained from R- and S-enantiomers of sec.-phenylethylalcohol and 2,6-dichloropurine. All derivatives were tested for inhibition with purified hexameric E. coli purine nucleoside phosphorylase (PNP). For analogues showing IC50 < 10 microM, the type of inhibition and inhibition constants were determined. In all cases the experimental data were best described by the mixed-type inhibition model and the uncompetitive inhibition constant, Kiu, was found to be several-fold lower than the competitive inhibition constant, Kic. This effect seems to be due to the 6-aryloxy- or 6-arylalkoxy substituent, because a natural PNP substrate adenine, as well as 2-chloroadenine, show mixed type inhibition with almost the same inhibition constants Kiu and Kic. The most potent inhibition was observed for 6-benzylthio-2-chloro-, 6-benzyloxy-2-chloro-, 2-chloro-6-(2-phenyl-1-ethoxy), 2-chloro-6-(3-phenyl-1-propoxy)- and 2-chloro-6 ethoxypurines (Kiu = 0.4, 0.6, 1.4, 1.4 and 2.2 microM, respectively). The R stereoisomer of 2-chloro-6-(1-pheny-1-ethoxy)purine has Kiu = 2.0 microM, whereas inhibition of its S counterpart is rather weak (IC50 > 12 microM). More rigid (e.g. phenoxy-), non-planar (cyclohexyloxy-), or more bulky (2,4,6 trimethylphenoxy-) substituents at position 6 of the purine base gave less potent inhibitors (IC50 = 26, 56 and > 100 microM, respectively). The derivatives are selective inhibitors of hexameric "high-molecular mass" PNPs because no inhibitory activity vs. trimeric Cellulomonas sp. PNP was detected. By establishing the ligand-dependent stabilization pattern of the E. coli PNP it was shown that the new derivatives, similarly as the natural purine bases, are able to form a dead-end ternary complex with the enzyme and orthophosphate. It was also shown that the derivatives are substrates in the reverse synthetic direction catalyzed by E. coli PNP. PMID- 10685498 TI - Bleomycin-induced DNA damage and DNA repair in chicken embryo cells as compared to X-irradiation. AB - Following in vitro- and in ovo-exposure of chicken embryo cells, the level of bleomycin (BM)-induced damage was evaluated by using DNA synthesis, nucleoid sedimentation (SED), and viscometry of alkaline cell lysates (VISC). This damage was compared to X-irradiation, using 5.9-378 nM BM in vitro, 1.5-116 micrograms BM/egg in ovo, and 2-32 Gy, respectively, in vitro as well as in ovo. With respect to BM, the most notable result is the increase in DNA synthesis and VISC at the lowest concentrations of the drug. A decrease in both parameters was observed at high BM concentrations and following exposure to X-rays, concomitantly with an increase in SED. Regarding the radiomimetic drug BM and X rays, different modes of DNA damage and DNA repair are suggested by previous investigations and the present results. Therefore, further evidence is presented, that the chicken embryo can act as a simple, rapid and inexpensive test system to characterize the biological effects of many nucleo- and/or cytotoxic agents. PMID- 10685499 TI - 1H-cyclopenta[b]benzofuran lignans from Aglaia species inhibit cell proliferation and alter cell cycle distribution in human monocytic leukemia cell lines. AB - Thirteen naturally occurring 1H-cyclopenta[b]benzofuran lignans of the rocaglamide type as well as one naturally occurring aglain congener all of them isolated from three Aglaia species (Aglaia duperreana, A. oligophylla and A. spectabilis) collected in Vietnam were studied for their antiproliferative effects using the human monocytic leukemia cell lines MONO-MAC-1 and MONO-MAC-6. Only rocaglamide type compounds showed significant inhibition of [3H-]thymidine incorporation and the most active compound didesmethylrocaglamide inhibited cell growth in a similar concentration range as the well-known anticancer drug vinblastine sulfate. Detailed structure-activity analysis indicated that the OH group at C-8b which is a common structural feature of most naturally occurring rocaglamide compounds is essential for the described antiproliferative activity since replacement of this group by methylation led to a complete loss of the inhibitory activity for the resulting derivative. Rocaglamide derivatives rapidly inhibited DNA as well as protein biosynthesis of MONO-MAC-6 cells at concentrations well below those of actinomycin D or cycloheximide which were used as positive controls in the respective experiments. Didesmethylrocaglamide was furthermore able to induce growth arrest of MONO-MAC-1 cells in the G2/M and probably G0/G1-phase of the cell cycle with no morphological indication of cellular damage. Our data suggests that 1H-cyclopenta[b]benzofuran lignans of the rocaglamide type act primarily by a cytostatic mechanism. PMID- 10685500 TI - Carboxymethylated glucan inhibits lipid peroxidation in liposomes. AB - Protective capabilities were studied of carboxymethylated (1-->3)-beta-D-glucan from Saccharomyces cerevisiae cell wall against lipid peroxidation in phosphatidylcholine liposomes induced by OH radicals produced with Fenton's reagent (H2O2/Fe2+) and also by microwave radiation using absorption UV-VIS spectrophotometry. A significant decrease in the conjugated diene production, quantified as Klein oxidation index, was observed in the presence of a moderate amount of added glucan. Increase of the oxidation index was accompanied with enhanced carboxyfluorescein leakage as a result of liposome membrane destabilization. This process was markedly suppressed with glucan present in the liposome suspension. Therefore, glucan may be considered as a potent protector against microwave radiation-induced cell damage. PMID- 10685501 TI - The anticancer drug chlorambucil interacts with the human erythrocyte membrane and model phospholipid bilayers. AB - The plasma membrane has gained increasing attention as a possible target of antitumor drugs. It has been reported that they act as growth factor antagonists, growth factor receptor blockers, interfere with mitogenic signal transduction or exert direct cytotoxic effects. Chlorambucil (4-[p-(bis[2 chloroethyl]amino)phenyl]butyric acid) is an alkylating agent widely used in the treatment of chronic lymphocytic leukaemia. Contradictory reports have been published concerning its interaction with cell membranes. Whereas a decrease in the fluidity of Ehrlich ascite tumor cells has been adduced, no evidences were found that chlorambucil changes membrane lipid fluidity and alkylating agents had effects in these systems even at highly toxic concentrations. Our results showed that chlorambucil at a dose equivalent to its therapeutical concentration in the plasma (3.6 microM) caused the human erythrocyte membrane to develop cup-shaped forms (stomatocytes). Accordingly to the bilayer couple hypothesis, this means that the drug is inserted into the inner monolayer of the erythrocyte membrane, a conclusion supported by X-ray diffraction performed on multilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), representative of phospholipid classes located in the outer and inner monolayers of the erythrocyte membrane, respectively. It is concluded that the cytotoxic effect of chlorambucil might be due to alteration of the structure and therefore of the physiological properties of cell membranes such as fluidity, permeability, receptor and channel functions. PMID- 10685502 TI - The activity of thymidine phosphorylase correlates with tumor size and lymph nodes status in breast carcinoma. AB - The platelet-derived endothelial cell growth factor (PD-ECGF) is one of the potent angiogenic factors. Recently, its homology with thymidine phosphorylase (dThdPase), an enzyme involved in pyrimidine nucleoside metabolism, has been shown. In the present study, dThdPase activity was evaluated spectrophotometrically in 43 breast carcinomas and in 19 cases of non-neoplastic breast tissues. The mean dThdPase activity in breast cancer was almost six fold higher than in normal, non-neoplastic breast tissues (1.92 and 0.29 mumol thymine (T) x mg prot.-1 x h-1 respectively). The enzyme activity significantly correlated with axillary lymph node status (p = 0.0076) and with tumor size (p = 0.0099). Besides, the intratumoral microvessel density (MD) was evaluated using the CD 31 mouse anti-human monoclonal antibody, and there was no correlation between the level of enzymatic activity and a number of microvessels. The positive significant correlation of thymidine phosphorylase activity with prognostic factors in breast cancer patients with no relation to the number of microvessels needs further examination to confirm the prognostic significance of the level of dThdPase. PMID- 10685503 TI - Scaling of some metabolic enzymes in liver of a freshwater teleost: an adaptive mechanism. AB - The activities of mitochondrial malate dehydrogenase (mMDH) and the total mitochondrial proteins increase as a function of body mass in the freshwater catfish, Clarias batrachus. It clearly indicates an increase in energy production in larger-sized individuals for various purposes including prey-predator interactions. The higher activity of lactate dehydrogenase (LDH) in larger fish may indicate more production of lactate for gluconeogenesis in the liver to meet emergency requirements of increased energy demand. However, the activity of cytoplasmic malate dehydrogenase (cMDH) decreases with the increasing body mass of the fish which reflects reduction in NADPH production and, in turn, reduced lipogenesis in liver of larger individuals. Thus, the present observations suggest an adaptive mechanism dealing with the higher energy budget, and reduced synthetic activities (lipogenesis) in the liver of larger-sized freshwater catfish. This type of biochemical scaling might be also supporting other metabolic pathways in order to adjust some physiological functions for survival in the aquatic environment. PMID- 10685504 TI - Contribution of a metal-peroxide adduct to neurodegeneration is due to its oxidative protease activity. AB - Many hypotheses have been developed to explain aging and age-related neurodegenerative disorders; one of the most compelling is the role of oxidative stress to induce changes in protease activity in brains of patients of Alzheimer's disease and prion disease. At the moment however, there is no clear answer how protein degradation may be achieved in the brain. We have observed that several metal compounds can degrade proteins in the presence of hydrogen peroxide, and elucidated the reaction scheme based on the new theoretical point for the reactivity of a metal-peroxide adduct with eta 1-coordination mode. In this article we would like to point out the importance of a copper(II)-peroxide adduct to promote neurodegenerative diseases such as prion disease and amyotrophic lateral sclerosis through its oxidative protease function. PMID- 10685505 TI - Androconial hairbrushes of the Syntomis (Amata) phegea (L.) group (Lepidoptera, Ctenuchinae): a synapomorphic character supported by sequence data of the mitochondrial 16S rRNA gene. AB - Males of several palaearctic Syntomis/Amata species (Lepidoptera: Arctiidae) possess androconial hairbrushes in connection with the foreleg coxa. The cuticular structure of these potentially behaviour-related and pheromone dissipating brushes is described. Such male-specific organs and signals play a crucial role in the female choice procedure. The presence of hairbrushes was found in 17 out of 28 inspected species of the tribe Syntomini. All members of the Syntomis phegea group (Europe to Central Asia, as well as Caspian, Caucasian and near-middle East species) have these structures, and only three oriental and south Asian, but none of three African species, carry this trait. The common genetic base of this morphological character is supported by an analysis of mitochondrial 16S rRNA from 19 representative taxa; species with hairbrushes form a monophyletic clade and the brushes are a synapomorphic character. This genetic finding corroborates the ethological significance of these organs. Phylogenetic data show a substantial genetic divergence between the tribe Ctenuchini (New World species) and the Old World Syntomini. Furthermore, DNA sequence data suggest a split of the genus Amata (sensu Obraztsov, 1966) in two distinct genera, Amata (without hairbrushes) and Syntomis (with hairbrushes). PMID- 10685506 TI - Flavonoids and terpenoids from Luma gayana (Barn.) Burret. AB - The flavonoids 5-hydroxy-7-methoxyflavanone, 6,8-dimethyl-5,7-dihydroxyflavanone and 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone, a mixture of alkyl esters of p-coumaric acid, the triterpenoids oleanolic acid and maslinic acid, the monoterpenoid 1 alpha,2 beta,4 beta-trihydroxy-p-menthane, the sesquiterpenoid clovandiol and beta-sitosterol were isolated from the aerial parts of Luma gayana (Barn.) Burret. This is the first report on the chemistry of this species. PMID- 10685507 TI - Possible involvement of beta-adrenergic receptors in the enhancement of nocturnal pineal N-acetyltransferase activity due to parathion administration. AB - The purpose of the present study was to examine the effects of administration of sublethal doses of O,O-diethyl-O-p-nitrophenyl phosphorothioate (parathion) on serum epinephrine (EPI) and norepinephrine (NE), as well as on night-time rat pineal melatonin synthesis, both in the presence and absence of propranolol, a beta-adrenergic receptor antagonist. In the first experiment, two groups of adult albino rats were administered parathion orally (1.08 and 2.17 mg/kg/day; the total received by each animal was 6.5 and 13.0 mg/kg body weight over 6 days); another two groups received corn oil only. Animals were killed at 23:00 and 01:00 h by decapitation. Serum EPI was augmented at 01:00 h, but NE was increased at 01:00 and 23:00 h due to administration of the high dose of parathion (13 mg/kg). In the second experiment, two groups of adult male albino rats were administered parathion orally (13 mg/kg); another two groups received an intraperitoneal injection of propranolol (20 mg/kg body weight, 1 h before the lights were turned off). In addition, two groups were given a saline injection. Four hours after darkness onset, pineal N-acetyltransferase (NAT) activity as well as pineal and serum melatonin levels were measured. Parathion by itself significantly augmented nocturnal pineal NAT activity and serum melatonin levels in otherwise untreated rats; the insecticide was ineffective in reference to this enzyme when it was given in conjunction with the beta-adrenergic receptor antagonist propranolol. The augmentation of NAT activity by parathion also caused significant reduction in pineal serotonin (5-HT); again, this response was blocked by propranolol treatment. The results are consistent with the idea that parathion influences pineal 5-HT metabolism either at the level of the beta-adrenergic receptor or via the sympathetic innervation to the pineal gland. PMID- 10685508 TI - Hepatotoxicity of tetrabromobisphenol-A: effects of repeated dosage in rats. AB - Tetrabromobisphenol-A (TBBP-A) is used as a reactive flame retardant and as an intermediate in the production of other flame-retardants. In our study, TBBP-A was administered intragastrically, daily for 7 or 7-28 days at three dose levels. Significant changes of biochemical indicators were noted with regard to glutathione (GSH), malondialdehyde (MDA) and 5-aminolevulinate dehydratase (ALA D). The level of GSH was lowered by the two higher doses (female rats only) and MDA was elevated by the highest dose (male rats only). The ALA-D activity reacted in opposite directions for both sexes. Other indicators did not yield any conclusive results. The 28-day study was performed on female rats. For GSH and MDA the medium dose resulted in a systematic increase. Insignificant changes in ALA-D activity in the liver were observed throughout the experiment. The activity of 5-aminolevulinate synthase had a decreasing tendency at 250 mg/kg of TBBP-A during the whole time of observation. Other general indices such as the activity of gamma-glutamyltransferase, concentration of microsomal proteins and the level of cytochrome P-450 did not show any significant changes. The most pronounced changes were noted with regard to indicators of porphyrogenic action. The results suggest that TBBP-A is capable of disturbing the heme metabolism in rats. PMID- 10685509 TI - Effect of dietary aluminum on tissue nonheme iron and ferritin levels in the chick. AB - Aluminum toxicity is well documented but the mechanism of action is poorly understood. In renal failure patients with aluminum overload, disturbances in iron metabolism leading to anemia are apparent. Few animal models, however, have been used to study the effects of dietary aluminum on iron metabolism. The purpose of this study was to determine if dietary aluminum exposure alters tissue iron and ferritin concentrations in the chick, as has been found in cultured human cells exposed to aluminum. Groups of day-old chicks were fed purified diets containing one of two levels of iron (control or high iron), and one of three levels of aluminum chloride in a 2 x 3 factorial design. Diets were consumed ad libitum for 1 week, then pair-feeding was initiated for 2 more weeks. A seventh group consumed a low iron diet ad libitum for comparative purposes. After the 3 week feeding period, samples of kidney, liver, and intestinal mucosa were analyzed for nonheme iron and ferritin concentrations by a colorimetric assay and SDS-PAGE, respectively. Results showed that dietary aluminum intake reduced iron stores in liver and intestine, but had no effect on nonheme iron levels in the kidney. Ferritin levels were reduced by aluminum intake in all tissues studied. The decreases in tissue ferritin levels were proportionately more than the decreases in tissue nonheme iron levels. This resulted in increased nonheme iron to ferritin ratios that amounted to as much as 140 and 525% in kidney and intestine, respectively. These findings are consistent with the interpretation that, in the growing chick, dietary aluminum can inhibit iron absorption, disrupt the regulation of tissue ferritin levels by iron, and potentially alter the compartmentalization and protective sequestration of iron within cells. PMID- 10685510 TI - Chromium increases pancreatic metallothionein in the rat. AB - The ability of chromium (Cr) salts to increase metallothionein (MT) levels in rat liver, kidney and pancreas, and its relationship with the presence of toxic effects are reported here. Rats were injected subcutaneously with 0, 10, 20, 30, 40, or 50 mg K2Cr2O7/kg and sacrificed 24 h later. Total Cr accumulation followed a dose-dependent pattern, levels in kidney being higher than those in liver or pancreas, suggesting different tissue bioavailabilities and accumulation patterns. Cr(IV) administration resulted in a tissue-specific MT induction: pancreas and liver showed five- and 3.5-fold MT increases, respectively; no increase was observed in the kidney. A positive correlation was observed between zinc and MT concentrations in liver, and between total Cr and MT concentrations in pancreas. Serum alpha-amylase activity showed a dose-dependent increase starting from 20 mg/kg, whereas serum glucose levels increased at doses higher than 30 mg/kg. Serum aspartate aminotransferase and alanine aminotransferase activities were increased in a dose-dependent manner, from 20 and 30 mg/kg, respectively. Our results showed that treatment with Cr(VI) can induce MT synthesis in pancreas and suggests a subsequent binding of Cr to MT. Also, pancreas is a target organ for Cr toxicity, and the usefulness of alpha-amylase activity as a sensitive biomarker of Cr toxicity in human exposed populations merits further study. PMID- 10685511 TI - The effect of vitamin E exposure on cadmium toxicity in mouse embryo cells in vitro. AB - Heavy metals such as cadmium pose a number of environmental problems in addition to being detrimental to human health. Cadmium is known to be embryotoxic in animal models and to cause brain, limb and craniofacial malformations. Among numerous mechanisms proposed for cadmium toxicity are oxidative stress and lipid peroxidation. Vitamin E has been found to have antioxidant and cytoprotective properties in cultured cells but its effect on cadmium embryonic toxicity has not yet been determined. Epithelial-like cells derived from day-8 whole-mouse embryos were used as a model embryonic tissue. Cadmium toxicity in these cultured cells was found to be both time and concentration dependent. Prior exposure to 50 microM alpha-tocopherol or 25 or 50 microM alpha-tocopherol acetate resulted in a marked reduction in the toxicity of 5 microM CdCl2. The apparent cytoprotective effects may be partly non-specific, however, as a general growth enhancement was observed after vitamin E exposure in the absence of cadmium. PMID- 10685512 TI - Differential sensitivities of MRP1-overexpressing lung tumor cells to cytotoxic metals. AB - The human multidrug-resistance protein (MRP1), known to mediate cellular efflux of a wide range of xenobiotics, including anticancer drugs, has also been shown to transport antimony, thereby conferring resistance to this heavy metal. The aim of the present study was to investigate whether other cytotoxic metals could be handled by MRPI using MRP1-overexpressing lung tumor GLC4/Sb30 cells. Such cells were found to be 3.4-, 12.7- and 16.3-fold more resistant than parental GLC4 cells to mercuric ion, arsenite and arsenate, respectively, whereas they remained sensitive to other cytotoxic metals tested such as copper, chromium, cobalt or aluminium. MK571, a potent inhibitor of MRP1 activity, almost totally reversed resistance of GLC4/Sb30 cells to mercuric ions and arsenic while it did not significantly alter sensitivity of GLC4 cells to metals. Arsenate-treated GLC4/Sb30 cells were found to poorly accumulate arsenic through increased MK571 inhibitable efflux of the metal. Arsenate, however, failed to alter MRP1-mediated transport of known MRP1 substrates such as calcein and vincristine. In conclusion, these findings demonstrated that MRP1 likely handled some, but not all, cytotoxic metals such as arsenic and mercuric ions in addition to antimony, therefore resulting in reduced toxicity of these compounds towards MRP1 overexpressing cells. PMID- 10685513 TI - Early hepatic changes in rats induced by permethrin in comparison with DDT. AB - In this study permethrin [(3-phenoxyphenyl)-methyl-3-(2,2-dichloroethenyl)-2,2 dim ethylcyclopropanecarboxylate] and DDT [1,1-(2,2,2 trichloroethylidene)-bis-(4 chlorobenzene)] were compared in rats for their effects on early hepatic changes, proposed in the literature to be useful endpoints in screening for non-genotoxic hepatocarcinogenesis and/or liver tumour promotion. We compared the effects of both insecticides on the following endpoints: hepatomegaly, mitogenesis (DNA synthesis, mitotic activity, percentage of binuclear cells) and liver pathology. Male Wistar rats received permethrin (PERM) or DDT in one, three, five and 14 daily oral doses (at 24-h intervals) equivalent to 1/10 LD50. Distinct differences in early liver response between PERM and DDT were observed. DDT stimulated the early effect consisting of hepatomegaly accompanied by an increase in hepatocellular proliferation with signs of cell necrosis. Thus, it might be concluded, that the mitogenic effect of DDT was at least partly related to a regenerative liver response. Although PERM significantly affected DNA synthesis and increased binuclear hepatocytes, this compound did not increase the number of mitotic figures. These results suggest that PERM may inhibit of phase G2 in the cell cycle and consequently it may suppress the cell entering into the stage of mitosis (M-phase). In addition, the present findings provide evidence for the occurrence of abnormal mitoses in the hepatocytes of rats treated with DDT. PMID- 10685514 TI - Flow cytometry as a tool to monitor the disturbance of phagocytosis in the clam Mya arenaria hemocytes following in vitro exposure to heavy metals. AB - The effectiveness of toxicology biomonitoring programs could be improved by the addition of sensitive biomarkers. In this study the cell viability and sensitivity of phagocytic function of phagocytes from bivalves (Mya arenaria) to selected heavy metals were measured by flow cytometry, a novel approach. Hemocytes (phagocytes) collected from bivalves by puncture of the posterior adductor muscle were incubated in vitro for 18 h in hemolymph containing 10(-9) 10(-3)M of cadmium chloride, zinc chloride, mercuric chloride, methylmercury chloride or silver nitrate, before determining their capacity to phagocytose fluorescent latex beads by flow cytometry. Heterogeneity of the hemocyte cell population was determined by forward scatter (FSC) and side scatter (SSC) cytometric profile which showed two distinct cell populations. At low doses (10( 9), 10(-8) M), all the metal compounds studied stimulated phagocytic activity except silver nitrate. At higher levels of exposure (10(-6), 10(7) M), all metals caused a significant concentration-related decrease in hemocyte phagocytosis activity. From the concentration of each metal inducing 50% suppression (IC50) of the phagocytic activity, the immunotoxic potential of metals with respect to phagocytic function can be ranked in the following increasing order: ZnCl2 < CdCl2 < AgNO3 < HgCl2 < CH3HgCl. Parallel analysis of hemocyte viability showed that suppression of phagocytosis by heavy metals was not solely related to a decreased cell viability. These results reveal the high but different degree of sensitivity of the phagocytosis activity of bivalves with respect to heavy metals, as measured by flow cytometry, and demonstrate that flow cytometry is a potentially useful tool in ecotoxicological monitoring. PMID- 10685515 TI - Role of vaginal sonography and hysterosonography in the endoscopic treatment of uterine myomas. AB - OBJECTIVE: To summarize the advantages and disadvantages of the various imaging techniques used to evaluate uterine leiomyomas preoperatively and to propose a classification system for intramural and subserosal leiomyomas that may better serve the endoscopist in surgical treatment. DESIGN: A MEDLINE search of the available literature was performed. CONCLUSION(S): Selective use of the various imaging techniques is required based on the clinical situation. Classification systems that describe the degree of myometrial involvement are needed for appropriate case selection and counseling by the endoscopist. PMID- 10685516 TI - Altered gene expression and secretion of interleukin-6 in stromal cells derived from endometriotic tissues. AB - OBJECTIVE: To compare the expression of interleukin-6 (IL-6) in endometrial and endometriotic cells. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology, Tottori University Hospital, Yonago, Japan. PATIENT(S): Twenty patients who underwent either hysterectomy or laparoscopic surgery. INTERVENTION(S): Endometrial and endometriotic stromal cells were obtained from normal endometrium and from chocolate cyst linings of the ovary. Peritoneal macrophages were isolated from peritoneal fluids. Cells were cultured in the presence or absence of tumor necrosis factor-alpha. MAIN OUTCOME MEASURE(S): Gene expression of IL-6 was examined by Northern blot analysis. Interleukin-6 protein production was examined by immunocytochemical staining and ELISA. RESULT(S): A single IL-6 messenger RNA band of approximately 1.3 kilobases was detected in endometriotic stromal cells. Tumor necrosis factor-alpha increased the expression of IL-6 messenger RNA in endometriotic cells in a dose-dependent manner. In endometrial stromal cells, IL-6 messenger RNA signals were much weaker. Endometriotic stromal cells produced significantly larger amounts of IL-6 compared with endometrial stromal cells under basal conditions and after stimulation with tumor necrosis factor-alpha. Interleukin-6 protein was detected in cells isolated from endometriotic tissues by immunocytochemical staining. Interleukin-6 production by cultured macrophages from patients with endometriosis and endometriotic stromal cells was comparable. CONCLUSION(S): Altered gene expression and protein secretion of IL-6 in patients with endometriosis may contribute to the pathogenesis of the disease and/or to endometriosis-associated infertility. PMID- 10685517 TI - Interleukin-6: another piece of the endometriosis-cytokine puzzle. PMID- 10685518 TI - Embryo donation programs and policies in North America: survey results and implications for health and mental health professionals. AB - OBJECTIVE: To use survey results from Society of Assisted Reproductive Technology to describe program policies regarding embryo donation, report protocols used for the disposition of cryopreserved embryos, and discuss clarification of guidelines governing ethical and psychosocially informed embryo donation. METHOD(S): A 66 item questionnaire was sent to the 312 Society of Assisted Reproductive Technology programs, generating 108 responses. RESULT(S): Seventy-eight (72%) of 108 programs offer embryo donation. Forty (37%) have actually performed donation, with 246 cycles completed and 53 "take-home babies." Disposition agreements for donors address divorce (92%) and death (90%). Only 28% require that potential donors undergo psychologic evaluation. Ninety-five percent of programs do not compensate donors. Seventy-one percent require a complete medical and psychologic history and 10% require genetic karyotyping. Three percent limit the number of donations. Eligible recipients include married couples (100%), unmarried couples (61%), lesbian couples (55%), and single women (59%). Sixty-four percent of programs require psychologic screening. Storage limits range from 2-10 years. Forty-nine percent of programs have unclaimed embryos in storage. CONCLUSION(S): Embryo donation is more often contemplated than performed. Variability in program procedures and policies suggests that guidelines need to be clarified. The complexity of the psychosocial and ethical issues underscores the importance of a routine, comprehensive psychologic assessment. PMID- 10685519 TI - Semen quality of workers occupationally exposed to hydrocarbons. AB - OBJECTIVE: To determine whether occupational exposure of men to hydrocarbons has adverse effects on the quality of their semen. DESIGN: Comparative study. SETTING: The rubber industry in Mexico City. PATIENT(S): Forty-eight workers who were exposed to hydrocarbons for 2-24 years and 42 unexposed workers. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Environmental hydrocarbon concentrations were determined by continuous air monitoring in all areas of the factory. Analyses of semen samples were performed in accordance with World Health Organization criteria. RESULT(S): Hydrocarbon concentrations were as follows: ethylbenzene, 220.7-234 mg/m3; benzene, 31.9-47.8 mg/m3; toluene, 189.7-212.5 mg/m3; and xylene, 47-56.4 mg/m3. The number of subjects with ejaculates that had normal characteristics was greater in the unexposed group (76%) than in the exposed group (17%). More abnormal characteristics were found in the semen of exposed workers than unexposed workers, including alterations in viscosity, liquefaction capacity, sperm count, sperm motility, and the proportion of sperm with normal morphology. Some abnormal characteristics correlated with the number of years of exposure to the hydrocarbons. CONCLUSION(S): Damage to the spermatogenic process resulting from hydrocarbon exposure was demonstrated by an increased rate of abnormalities in the semen of exposed workers compared with unexposed workers. This information may be useful for conducting future analyses of reproductive risks related to exposure to high concentrations of hydrocarbons. PMID- 10685520 TI - Antisperm autoantibody response is reduced by early repair of a severed vas deferens in the juvenile rat. AB - OBJECTIVE: To determine whether antisperm autoantibody production after prepubertal vas injury is influenced by immediate repair of the vas compared to delay of the reanastomosis until sexual maturity. DESIGN: Animal study comparing early repair, late repair, and sham-operated groups. SETTING: Research laboratory in a medical school. PATIENT(S): Lewis rats. INTERVENTION(S): After division of the vas deferens in juvenile rats, animals in an early repair group had the vasa repaired immediately by using an absorbable intraluminal stent. Animals in a late repair group had vasa obstructed by ligation until after puberty, when they underwent microsurgical vasovasostomy (age 60 days). MAIN OUTCOME MEASURE(S): Antisperm antibodies were assayed by ELISA. The weights of reproductive organs were determined, and samples of testis were studied by light microscopy. RESULT(S): The antisperm antibody response was less when the vas was repaired immediately than if the repair was delayed until after puberty. There was a low incidence of testicular alteration in the repair groups and none in sham-operated animals. CONCLUSION(S): If the vas deferens is injured or obstructed prepubertally, there may be a benefit to considering immediate repair to reduce the likelihood of developing antisperm autoantibodies, which have been associated with reduced fertility. PMID- 10685521 TI - Effect of vasovasostomy on contralateral testicular damage associated with unilateral vasectomy in mature and immature Lewis rats. AB - OBJECTIVE: We sought to determine if laser-assisted vasovasostomy could reverse the contralateral histologic testicular changes associated with unilateral vasectomy. DESIGN: A prospective, randomized, blinded, controlled study. SETTING: Animal microsurgical laboratory, St. John's Mercy Medical Center, St. Louis, Missouri. PATIENT(S): Twenty mature and 20 immature male Lewis rats. INTERVENTION(S): Ten mature and 10 immature male Lewis rats underwent unilateral vasectomy. At 5 months, testicular biopsy and laser-assisted vasovasostomies were performed followed 2 months later by evaluation of vas patency and repeat testicular biopsy. Control animals consisted of 10 rats in each group, 5 that underwent sham operations and 5 that had halothane anesthesia alone. RESULT(S): In the immature and mature groups unilateral vasectomy resulted in marked contralateral testicular damage in 30% (3 of 10) and 50% (5 of 10), respectively. Vas patency determined 2 months after vasovasostomy was 80% (8 of 10) in the mature group and 89% (8 of 9) in the immature group. No animal that had contralateral testicular changes after vasectomy and a patent vas after vasovasostomy showed improvement in testicular histology. CONCLUSION(S): It appears that contralateral testicular damage associated with unilateral vasectomy is not improved 2 months after successful vasovasostomy in mature or immature Lewis rats. PMID- 10685522 TI - A novel, rapid, and accurate method for detecting microdeletion involving the DAZ gene in infertile men. AB - OBJECTIVE: To report on a novel, accurate method for detecting microdeletion involving the DAZ gene in infertile men. DESIGN: Retrospective clinical study. SETTING: University Infertility Center of Cochin Hospital, Paris, France. PATIENT(S): Infertile patients (n = 25) consulting our infertility department during 1998. The patient cohort included subjects with nonobstructive azoospermia and oligoasthenospermia. INTERVENTION(S): Blood samples were collected from each subject. MAIN OUTCOME MEASURE(S): DNA analysis using polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE). RESULT(S): We used a new molecular genetic strategy to rapidly identify deletions of the Y chromosome that include the DAZ locus. The experiment consists of amplifying simultaneously exon 4 of the DAZ and DAZLA genes with the use of specific primers that are complementary to intronic sequences of these genes. DGGE was used to separate the two PCR products, with good resolution. In infertile men with a microdeletion of the DAZ gene, this method allows amplification of an internal control when a deletion of that portion of the Yq chromosome is observed on a single amplification. CONCLUSION(S): This PCR-DGGE method for detection of DAZ gene deletion is simple and fast and does not require the use of radioactive elements. Compared with the classic PCR approach, this new method allows the amplification of the DAZLA copy to be used as an effective internal control in infertile men with microdeletion of the DAZ locus. This procedure could be particularly useful in screening for the DAZ locus in the diagnostic workup of nonobstructive azoospermia and severe oligoasthenoteratozoospermia. PMID- 10685523 TI - Herpes simplex virus infection of the uterine cervix--relationship with a cervical factor? AB - OBJECTIVE: To determine the prevalence of genital herpes simplex virus (HSV) in women of reproductive age and to evaluate a potential relation of asymptomatic HSV shedding with a cervical factor. DESIGN: Prospective study. SETTING: Outpatient infertility clinic of a university hospital. PATIENT(S): Randomly chosen asymptomatic women (n = 1,262) with a median age of 30 years. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Prevalence of cervical HSV, cervical index parameters, and other variables of CM quality, including CM penetrability in vivo and in vitro. RESULT(S): The prevalence of HSV infection of the uterine cervix was 5.2% (identified with cell culture). There was a tendency toward increased viscosity of the CM in HSV-positive women, but no significant relation with the other variables of CM quality (amount, spinnbarkeit, ferning, cervical appearance, and cellularity of the CM), or with the summarized Insler score or the cervical index according to World Health Organization guidelines. Postcoital testing and the in vitro penetration test, using either partners' or donors' semen, showed that the penetrability of the CM did not differ significantly between women with and without cervical HSV shedding. Asymptomatic cervical HSV infection was not significantly associated with bacterial colonization of the lower genital tract, with leukocyte counts in cervical secretions, with the pH of the CM or the vaginal fluid, or with antisperm antibodies in the CM. CONCLUSION(S): The results suggest that in asymptomatic women under controlled endocrine conditions, cervical HSV infection is not a significant cause of impaired quality and penetrability of the CM. PMID- 10685524 TI - Secondary amenorrhea and infertility caused by an inhibin-B-producing ovarian fibrothecoma. AB - OBJECTIVE: To report a case of secondary amenorrhea and infertility caused by an inhibin-B-producing ovarian fibrothecoma. DESIGN: Case report. SETTING: Academic medical center. PATIENT: A 37-year-old woman with a 2-year history of secondary amenorrhea and infertility. INTERVENTION(S): Operative removal of a 5-cm ovarian fibrothecoma. MAIN OUTCOME MEASURE(S): Luteinizing hormone, FSH, E2, inhibin-B, TSH, and prolactin measured preoperatively and postoperatively. Immunostaining of tumor cells for inhibin and LH. RESULT(S): Preoperative hormone levels were as follows: FSH, 1.7 mIU/mL; LH, 23.4 mIU/mL; E2, 31 pg/mL; and inhibin B, 1,154 pg/mL. Three weeks postoperatively, the FSH was 1.5 mIU/mL, LH decreased to 7.1 mIU/mL, E2 increased to 276 pg/mL, and inhibin-B decreased to 17 pg/mL. The fibrothecoma did not stain for LH but was strongly positive for inhibin. Regular menstrual cycles resumed 28 days postoperatively. CONCLUSION(S): Inhibin-B produced by an ovarian tumor profoundly suppressed FSH levels and resulted in secondary amenorrhea and infertility. Use of sensitive and specific immunoassays for inhibin-A and -B may aid in the differential diagnosis of hormonally active ovarian tumors. PMID- 10685525 TI - Influence of insulin resistance on total renin level in normotensive women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate the influence of insulin resistance on the plasma total renin level in normotensive women with polycystic ovary syndrome (PCOS). DESIGN: Prospective, controlled study. SETTING: University hospital. PATIENT(S): Twenty five normotensive women with PCOS were compared with 11 normotensive control women with regular cycles and no features of PCOS. INTERVENTION(S): Clinical, ultrasonographic, and hormonal findings were used to define PCOS. Insulin resistance was estimated by continuous infusion of glucose with model assessment in the early follicular phase. MAIN OUTCOME MEASURE(S): Plasma levels of total renin and angiotensin II and serum levels of gonadotropins, DHEAS, total T, free T, 17 alpha-hydroxyprogesterone, and PRL were determined. RESULT(S): Plasma concentrations of angiotensin II were similar in the PCOS group and the control group. The concentration of total renin in plasma was higher in women with PCOS than in healthy women independent of insulin resistance. The sensitivity and specificity of the plasma total renin level to diagnose women with PCOS were calculated as 80% and 71.4%, respectively. CONCLUSION(S): The plasma total renin level is higher in normotensive women with PCOS than in healthy women independent of insulin resistance. PMID- 10685526 TI - Laparoscopic treatment of polycystic ovaries with insulated needle cautery: a reappraisal. AB - OBJECTIVE: To evaluate the reproductive outcome and adhesion formation after a standardized laparoscopic treatment of polycystic ovary syndrome (PCOS) in clomiphene-resistant infertile women. DESIGN: Retrospective study. SETTING: University teaching hospital. PATIENT(S): One hundred twelve clomiphene-resistant anovulatory women with PCOS. INTERVENTION(S): Laparoscopic ovarian drilling using an insulated needle cautery. MAIN OUTCOME MEASURE(S): Ovulatory rate, pregnancy rate, and adhesion formation. RESULT(S): After surgery, ovulation occurred spontaneously in 73.2% of patients. The cumulative probability of conception at 12, 18, and 24 months after surgery was 54%, 68%, and 72%, respectively. With use of Cox's proportional hazards model, the effects of age, body mass index, and duration of infertility were evaluated. These factors were not associated with the pregnancy rate. Of 15 women who underwent a second-look laparoscopy, 11 women were found to be free of adhesions. Four women had periadnexal adhesions that were filmy, minimal, and found on the ovarian surface only. CONCLUSION(S): Laparoscopic ovarian drilling is an effective alternative treatment in clomiphene resistant anovulatory women with PCOS. The use of an insulated needle cautery is associated with a minimal amount of adhesion formation. PMID- 10685527 TI - Human chorionic gonadotropin as a predictor of outcome in assisted reproductive technology pregnancies. AB - OBJECTIVE: To determine whether serum hCG and progesterone levels obtained 16 days after ovulation are reliable predictors of pregnancy outcome. DESIGN: A retrospective study. SETTING: The data were obtained from two integrated Adelaide based clinics: the Queen Elizabeth Hospital and Wakefield Clinic. PATIENT(S): Women who have achieved a pregnancy through ART treatment. MAIN OUTCOME MEASURE(S): Analysis of data using logistic regression (STATA v.5.0) to predict a binary outcome: ongoing pregnancy or miscarriage. Ongoing pregnancy was defined as progression to >20 weeks' gestation. Miscarriage included spontaneous abortion, biochemical and ectopic pregnancies, and blighted ovum. RESULT(S): Human chorionic gonadotropin was found to be the main determinant of ongoing pregnancy. Age and progesterone had minor effects, whereas stimulation, luteal support, and treatment types were nonpredictive. Low hCG levels between 25 and 50 IU/L are associated with a low probability of ongoing pregnancy (<35%), whereas levels of >500 IU/L predict a >95% chance of ongoing pregnancy. CONCLUSION(S): A single serum hCG level 16 days after ovulation provides a useful predictor of pregnancy outcome. PMID- 10685528 TI - Human chorionic gonadotropin administration does not increase plasma androgen levels in patients undergoing controlled ovarian hyperstimulation. AB - OBJECTIVE: To investigate the effects of hCG administered to patients undergoing controlled ovarian hyperstimulation on levels of ovarian hormones, including androgens. DESIGN: Prospective analysis. SETTING: Assisted Reproduction Unit, Hopital Antoine Beclere, Clamart, France. PATIENT(S): Six infertile, normally ovulating volunteers. INTERVENTION(S): The women underwent controlled ovarian hyperstimulation with a GnRH agonist and hMG for IVF-ET. After the i.m. administration of hCG (10,000 IU), blood samples were drawn every 6 hours for 4 days. MAIN OUTCOME MEASURE(S): Plasma androstenedione, testosterone, progesterone, and E2 profiles. RESULT(S): Treatment with hMG increased plasma androstenedione and testosterone levels 1.4-fold and 2.6-fold, respectively. The administration of hCG did not increase plasma androstenedione and testosterone levels any further; mean daily levels remained at 2.3 ng/mL and 0.64 ng/mL, respectively. Circadian changes in androstenedione levels were evident after hCG administration. Plasma progesterone levels neared 10 ng/mL 19 hours after hCG administration, plateaued for 24 hours, and increased again thereafter. Plasma E2 levels declined during the first 2 days after hCG administration and then increased, concomitant with the second phase of progesterone elevation. CONCLUSION(S): In patients undergoing controlled ovarian hyperstimulation, androgen levels increased in response to hMG treatment, but no further elevation occurred after hCG administration. The rate of elevation of progesterone levels and the absolute levels achieved were 3-fold and 10-fold higher, respectively, than those observed during spontaneous menstrual cycles. PMID- 10685529 TI - Mutation analysis of the follicle-stimulating hormone receptor gene in girls with gonadotropin-independent precocious puberty resulting from autonomous cystic ovaries. AB - OBJECTIVE: To search for germline activating mutations of the FSH receptor in girls with gonadotropin-independent precocious puberty. DESIGN: Molecular studies in human tissue. SETTING: Four girls with polycystic ovaries and gonadotropin independent isosexual precocious puberty without clinical and molecular features of McCune-Albright syndrome. INTERVENTION(S): Peripheral blood was used for DNA extraction. The alpha-subunit of the Gs gene and the entire exon 10 of FSH receptor gene were amplified by polymerase chain reaction (PCR). Gs-alpha mutations characteristic of McCune-Albright syndrome were excluded by denaturating gradient gel electrophoresis (DGGE) and allele-specific PCR. Exon 10 of the FSH receptor gene was analyzed by DGGE and direct sequencing. MAIN OUTCOME MEASURE(S): Results of DGGE and direct sequencing. RESULT(S): No germline activating mutations were detected in exon 10 of our patients. Instead, two previously described polymorphisms were found, leading to the substitution of alanine for threonine at position 307 and of serine for asparagine at position 680 of the FSH receptor molecule. CONCLUSION(S): Germline activating mutations were not found in exon 10 of the FSHR gene in any of our patients. Further studies, preferably in ovarian tissue, will be required to exclude the presence of somatic activating mutations of the FSH receptor in these patients. PMID- 10685530 TI - Successful induction of ovulation using highly purified follicle-stimulating hormone in a woman with Kallmann's syndrome. AB - OBJECTIVE: To describe a woman with Kallmann's syndrome who was treated successfully with highly purified FSH to achieve ovulation induction and pregnancy. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 32 year-old woman with Kallmann's syndrome who had been treated with oral contraceptives to prime secondary sex characteristics and genital organs since the age of 16 years. INTERVENTION(S): Highly purified FSH was administered intramuscularly for a total dose of 3,825 IU. MAIN OUTCOME MEASURE(S): Follicle number and diameter. RESULT(S): Three follicles with a diameter of > 1.7 cm and an endometrial thickness of 8 mm were observed. A clinical pregnancy, which subsequently was spontaneously aborted, was obtained. CONCLUSION(S): In primed patients with Kallmann's syndrome, highly purified FSH may be a useful alternative to pulsatile GnRH or menopausal gonadotropins to achieve ovulation induction and pregnancy. PMID- 10685531 TI - Effects of clomiphene citrate on the endometrium of regularly cycling women. AB - OBJECTIVE: To study the effects of clomiphene citrate (CC) on the endometrium of regularly cycling women. DESIGN: Prospective, controlled study. SETTING: Department of Obstetrics and Gynaecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand. PATIENT(S): Thirty healthy, regularly cycling, female volunteers. INTERVENTION(S): All volunteers were studied for two consecutive cycles, one control cycle and one CC-treated cycle. Clomiphene citrate (100 mg/d) was given on days 3-7 of the CC-treated cycles. Ultrasonography was performed daily to assess ovulation. Ultrasonography and endometrial biopsy were performed, and blood samples were obtained for determination of E2 and progesterone levels 7 days after ovulation in both the control and CC-treated cycles. MAIN OUTCOME MEASURE(S): Histologic dating, morphometric analysis, and ultrasonographic appearance and thickness of the endometrium. RESULT(S): Histologic dating and ultrasonographic appearance and thickness of the endometrium were similar in the control and CC-treated cycles, but morphometric parameters were different. The number of glands per square millimeter and the mean diameter of the glands were lower in the CC-treated cycles than in the control cycles, but the number of vacuolated cells per 1,000 glandular cells was higher. CONCLUSION(S): Clomiphene citrate has effects on the endometrium of regularly cycling women, as demonstrated by a reduction in glandular density and an increase in the number of vacuolated cells. PMID- 10685532 TI - Are randomized trials of hormone replacement therapy in symptomatic women with breast cancer feasible? AB - OBJECTIVE: To evaluate the feasibility of conducting a large randomized trial of HRT in symptomatic women with early-stage breast cancer. DESIGN: Open randomized study. SETTING: Outpatient clinics at The Royal Marsden and St. George's Hospitals, London. PATIENT(S): One hundred postmenopausal women with early-stage breast cancer, experiencing vasomotor symptoms and/or vaginal dryness. INTERVENTION(S): Randomization (1:1) to HRT or no HRT for 6 months. MAIN OUTCOME MEASURE(S): Acceptance, continuance rates, and the reasons eligible women declined study entry. RESULT(S): Acceptance (38.8%) and continuance rates (>80%) were encouraging. The efficacy of HRT did not appear to be antagonized with concomitant tamoxifen. Seventy-five percent of women continued HRT after the study ended. Three women developed metastatic disease. Two used HRT. CONCLUSION(S): Despite informed consent, a national UK randomized trial of HRT should be feasible and has now been planned. Successful implementation necessitates the provision of information about HRT and the estrogen deficiency side effects of breast cancer therapy to health professionals and women with breast cancer. PMID- 10685533 TI - Embryonic karyotype of abortuses in relation to the number of previous miscarriages. AB - OBJECTIVE: To examine the frequency of chromosomal abnormalities in products of conception from patients with recurrent miscarriages in relation to the number of previous miscarriages. DESIGN: Retrospective analysis. SETTING: Nagoya City University Medical Hospital. PATIENT(S): A total of 1,309 women with a history of 2-20 consecutive first-trimester abortions. INTERVENTION(S): Chromosomal analysis performed on products of conception with use of a standard G-banding technique. MAIN OUTCOME MEASURE(S): The frequencies of abnormal and normal embryonic karyotypes for each number of previous abortions were studied. The subsequent pregnancy outcome of patients whose previous miscarriages were karyotyped were studied along with the predictive value of karyotyping of previous miscarriages for subsequent miscarriages. RESULT(S): The miscarriage rate increased with the number of previous spontaneous abortions. The frequency of abnormal embryonic karyotypes significantly decreased and that of normal embryonic karyotypes significantly increased with the number of previous abortions. Among 71 patients whose embryonic karyotypes were normal, 44 aborted subsequently, and 23 of 60 patients whose embryonic karyotypes were abnormal aborted subsequently. Patients with a previous normal embryonic karyotype aborted more frequently than those with an abnormal karyotype. CONCLUSION(S): The frequency of normal embryonic karyotypes significantly increases with the number of previous abortions, and a normal karyotype in a previous pregnancy is a predictor of subsequent miscarriage. PMID- 10685534 TI - Alterations in humoral immune responses associated with recurrent pregnancy loss. AB - OBJECTIVE: To investigate the reactivity of maternal antibodies with endometrium derived antigens and to correlate their association with recurrent pregnancy loss (RPL). DESIGN: Prevalence study. SETTING: Academic research center. PATIENT(S): Nulliparous women (n = 10), women with RPL (n = 15), pregnant women (n = 8), and multiparous women with a normal obstetric history (n = 20). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Reactive antibodies were analyzed by Western immunoblot techniques and quantitated by densitometry. RESULT(S): Antibodies from women with RPL and multiparous women recognized antigens ranging from 10-120 kd on normal endometrium and endometrial tumors. Antibodies from most women with RPL (10/15) and from multiparous women (15/20) recognized 65-kd and 80-kd proteins in normal endometrium. Antibodies from women with RPL recognized 21-kd and 28-kd antigens (12/15 and 13/15, respectively) in endometrial tumors at a significantly greater rate (than did antibodies from multiparous women (5/20 and 8/20, respectively). Women with RPL had significantly lower levels of asymmetric IgG compared with controls. CONCLUSION(S): Recurrent pregnancy loss may be linked with the failure to elicit asymmetric IgG and a unique immunologic recognition of endometrial antigens. PMID- 10685535 TI - Prospective, randomized, controlled study of in vitro fertilization-embryo transfer with a single dose of a luteinizing hormone-releasing hormone (LH-RH) antagonist (cetrorelix) or a depot formula of an LH-RH agonist (triptorelin). AB - OBJECTIVE: To confirm the value of a single dose of 3 mg of cetrorelix in preventing the occurrence of premature LH surges. DESIGN: Multicenter randomized, prospective study. SETTING: Reproductive medicine units. PATIENT(S): Infertile patients undergoing ovarian stimulation for IVF-ET. INTERVENTION(S): A single dose of 3 mg of cetrorelix (Cetrotide; ASTA Medica, Frankfurt, Germany) (115 patients) was administered in the late follicular phase. A depot preparation of triptorelin (Decapeptyl; Ipsen-Biotech, Paris, France) was chosen as a control agent (39 patients). Ovarian stimulation was conducted with hMG (Menogon; Ferring, Kiel, Germany). MAIN OUTCOME MEASURE(S): Premature LH surges (LH level >10 IU/L), progesterone level greater than 1 ng/L, and IVF results. RESULT(S): No LH surge occurred after cetrorelix administration. The patients in the cetrorelix group had a lower number of oocytes and embryos. The percentage of mature oocytes and fertilization rates were similar in both groups, and the pregnancy rates were not statistically different. The length of stimulation, number of hMG ampules administered, and occurrence of the ovarian hyperstimulation syndrome were lower in the cetrorelix group. Tolerance of cetrorelix was excellent. CONCLUSION(S): A cetrorelix single-dose protocol prevented LH surges in all patients studied. It compares favorably to the "long protocol" and could be a protocol of choice in IVF-ET. PMID- 10685536 TI - Preferred treatment of infertile women older than 37 years of age who demonstrate premature luteinization in the first evaluation cycle. AB - OBJECTIVE: To evaluate the efficacy of various treatments in abolishing premature luteinization in infertile women over 37 years old who are undergoing ovulation induction. DESIGN: Prospective, nonrandomized study. SETTING: Tertiary care medical clinic. PATIENT(S): Seventeen infertile women >37 years old in whom premature luteinization was detected during their evaluation (pretreatment) cycle. INTERVENTION(S): The patients underwent three consecutive treatment cycles with clomiphene citrate (group A), hMG (group B), and a GnRH agonist plus hMG (group C). MAIN OUTCOME MEASURE(S): Premature luteinization, defined as a progesterone/E2 ratio of >1 on the day of hCG administration. RESULT(S): Fifteen (88%) of the 17 patients in group A and 13 (76%) of the 17 patients in group B demonstrated premature luteinization. In contrast, only 1 (6%) of the 17 patients in group C had a progesterone/E2 ratio of >1 on the day of hCG administration. The mean (+/-SD) E2 level on the day of hCG administration was significantly higher in group C (1.236 +/- 772.7 pg/mL) than in group A (214.02 +/- 104.46 pg/mL) or group B (412.5 +/- 337 pg/mL). CONCLUSION(S): Pituitary desensitization with a GnRH agonist in conjunction with hMG may be of benefit for older infertile women who demonstrate early luteinization in their first evaluation cycle. PMID- 10685538 TI - Antiphospholipid antibodies and in vitro fertilization success: a meta-analysis. AB - OBJECTIVE: To evaluate whether the presence of antiphospholipid antibodies among women undergoing IVF affects the likelihood of IVF success. DESIGN: A meta analysis of seven eligible studies on antiphospholipid antibodies and IVF outcome. MAIN OUTCOME MEASURE(S): Odds ratios (ORs) and 95% confidence intervals (CIs) of an association between the presence of antiphospholipid antibodies and both clinical pregnancy and live birth from IVF. RESULT(S): There was no significant association between antiphospholipid abnormalities and either clinical pregnancy (OR 0.99; 95% CI 0.64-1.53) or live birth (OR 1.07; 95% CI 0.66-1.75) in IVF patients. CONCLUSION(S): The measurement of antiphospholipid antibodies is not warranted in patients undergoing IVF. PMID- 10685537 TI - The significance of delayed suppression using buserelin acetate and recombinant follicle-stimulating hormone in a long protocol in vitro fertilization program. AB - OBJECTIVE: To determine whether the time taken to achieve ovarian suppression has an impact on ovarian responsiveness and the outcome of IVF-ET. DESIGN: Retrospective analysis. SETTING: An assisted reproduction unit at a university center. PATIENT(S): Patients undergoing a long protocol of IVF-ET that included buserelin acetate therapy initiated on day 2 of the cycle and recombinant FSH. INTERVENTION(S): Patients were divided into two groups according to the duration of buserelin acetate therapy required to achieve pituitary and ovarian suppression (group 1 = 2 weeks, n = 172; group 2 = > or =3 weeks, n = 337). MAIN OUTCOME MEASURE(S): Number of recombinant FSH ampules administered, duration of ovarian stimulation (days), ovarian response, and IVF outcome. RESULT(S): The patients in group 2 had lower mean E2 levels after 5 days and 9 days of stimulation than the patients in group 1. The number of recombinant FSH ampules administered and the number of days of stimulation required were higher in group 2 than in group 1. These differences were prominent in the subgroups of older patients (> or =36 years) and patients who had no evidence of polycystic ovaries on ultrasound examination. The number of oocytes retrieved and fertilized, the cancelation rate, and the pregnancy rate were similar in the two groups. CONCLUSION(S): Prolonged administration of a GnRH agonist to achieve suppression leads to a reduced ovarian response, particularly in women > or =36 years of age, but does not affect the success rate of IVF-ET. PMID- 10685539 TI - Treatment of patients with long-standing unexplained subfertility with in vitro fertilization. AB - OBJECTIVE: To evaluate whether IVF is an effective treatment for long-standing unexplained subfertility. DESIGN: Retrospective cohort study. SETTING: Tertiary care infertility center in a university hospital. PATIENT(S): Two hundred two couples with unexplained subfertility of 2 years' duration or more who attended the center for their first IVF attempt. INTERVENTION(S): Couples were placed on a waiting list for IVF. They received no treatment until IVF was started. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR) while on the waiting list and PR after IVF treatment. RESULT(S): Complete data sets were available for 131 couples. Seventeen of 131 women became pregnant while waiting for IVF treatment (PR 0.9% per exposure cycle), whereas 45 of 119 receiving IVF treatment became pregnant (PR 17% per IVF attempt). CONCLUSION(S): IVF treatment has substantial added value over waiting and is an efficient treatment for long-standing unexplained subfertility. PMID- 10685540 TI - Follicle-stimulating hormone or human menopausal gonadotropin for ovarian stimulation in in vitro fertilization cycles: a meta-analysis. AB - OBJECTIVE: To reanalyze the results of using FSH alone and hMG during IVF treatment, taking into account the different protocols of administration of superactive GnRH agonist analogs. DESIGN: Meta-analysis. SETTING: The London Women's Clinic. PATIENT(S): Women undergoing IVF treatment. INTERVENTION(S): A meta-analysis of published randomized controlled trials from 1985 to 1999 of the use of FSH versus hMG for ovarian stimulation during IVF treatment. The common Peto odds ratio was calculated with use of a fixed effect model. The overall log odds ratio was estimated after demonstrating the consistency or homogeneity of the study results. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate per cycle of IVF. RESULT(S): The results suggested that in the "long and short GnRH agonists protocol" of IVF, FSH, and hMG were equally effective in achieving ovarian stimulation, and there were no differences in the clinical pregnancy rates per cycle of IVF. However, in protocols where no pituitary desensitization was used, FSH alone was more efficacious. CONCLUSION(S): The optimum choice of gonadotropin preparation for ovarian stimulation during IVF treatment is influenced by the regimen of pituitary desensitization used. The optimum gonadotropin to be used when GnRH antagonists are used has yet to be determined. PMID- 10685541 TI - Moderate and severe endometriosis is associated with alterations in the cell cycle of granulosa cells in patients undergoing in vitro fertilization and embryo transfer. AB - OBJECTIVE: To determine whether folliculogenesis is impaired in patients with endometriosis. DESIGN: Prospective study. SETTING: Yamagata University School of Medicine, Yamagata, Japan. PATIENT(S): Thirty women participating in an IVF program. INTERVENTION(S): The patients were divided into four groups according to the cause of their infertility: tubal factor (T), n = 7; male factor (M), n = 7; idiopathic (I), n = 7; and endometriosis (E), n = 9. Granulosa cells were obtained from the follicular fluid of each patient and analyzed by flow cytometry. MAIN OUTCOME MEASURE(S): The percentage of granulosa cells in each cell-cycle stage. RESULT(S): The mean (+/- SD) rate of apoptosis in the granulosa cells obtained from the patients with endometriosis was the highest among the four groups (T = 11.7% +/- 3.3%; M = 5.6% +/- 3.8%; I = 9.6% +/- 5.1%; and E = 18.6% +/- 9.6%). The percentage of S-phase granulosa cells was significantly higher in the patients with endometriosis than in all the other patients combined (E = 12.5% +/- 6%; T + M + I = 9.3% +/- 2.9%). The percentage of G2/M-phase granulosa cells was significantly lower in the endometriosis group than in the other three groups combined (E = 2.3% +/- 2.5%; T + M + I = 4.6% +/- 2.1%). CONCLUSION(S): Endometriosis impairs the cell cycle in granulosa cells. This phenomenon may have a detrimental effect on folliculogenesis. PMID- 10685542 TI - Images in reproductive medicine. Ultrastructural defects in acquired immotile sperm flagellae. PMID- 10685543 TI - Evaluation of the antifertility effect of magainin-A in rabbits: in vitro and in vivo studies. AB - OBJECTIVE: To evaluate the safety and contraceptive potential of magainin-A in rabbits. DESIGN: Controlled laboratory study. SETTING: Department of Immunology, Institute for Research in Reproduction, Mumbai, Parel, India. ANIMAL(S): Forty eight female New Zealand white rabbits. INTERVENTION(S): The effect of magainin-A on sperm motility (in vitro and in vivo studies) and on vaginal epithelium (histologic study) was assessed along with serum and liver biochemical profiles. MAIN OUTCOME MEASURE(S): Suitability of magainin-A for contraceptive use. RESULT(S): Magainin-A arrested sperm motility, and 1 mg of magainin-A administered intravaginally blocked conception. No histopathologic abnormalities in the vaginal tissue or any changes in serum biochemical profiles were observed. CONCLUSION(S): Magainin-A may be used as an effective and safe intravaginal contraceptive compound that also has antibacterial properties. PMID- 10685544 TI - Phase-dependent influence of nonsteroidogenic cells on steroidogenesis and prostaglandin production by the human corpus luteum. AB - OBJECTIVE: To test the hypothesis that paraluteal cells in the human corpus luteum (CL) modulate steroidogenesis and prostaglandin production by the CL. DESIGN: In vitro cell culture study using human luteal cells. SETTING AND PATIENT(S): Women (n = 7) with normal menstrual cycles who were undergoing operations for benign, nonovarian conditions during the midluteal phase (5-9 days after ovulation) or the late luteal phase (10-14 days after ovulation) at a university hospital. INTERVENTION(S): Steroidogenic and nonsteroidogenic human CL cells were isolated by mechanical and enzymatic digestion and density sedimentation. The cells were cultured (75,000 cells per well) for 24 hours either as a crude sample of all CL cells or as an enriched fraction of steroidogenic CL cells. MAIN OUTCOME MEASURE(S): Levels of progesterone, E2, prostaglandins F2alpha, E2, and I2 in conditioned medium. RESULT(S): Higher concentrations of progesterone, E2, and prostaglandins F2alpha, E2, and I2 were released into the media of the crude sample of all CL cells than into the enriched fraction of steroidogenic CL cells from the midluteal phase. No such difference was noted in CL cells from the late luteal phase. CONCLUSION(S): The paraluteal cells in the human CL stimulated progesterone and E2 synthesis. This may be mediated by an increase in prostaglandin production in the midluteal phase. PMID- 10685545 TI - Nucleoli in a pronuclei-stage mouse embryo are represented by major satellite DNA of interconnecting chromosomes. AB - OBJECTIVE: To investigate the arrangement of chromosomes within pronuclei-stage mouse zygotes. DESIGN: In vitro study. SETTING: Academic medical center. PATIENT(S): None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Location of major alpha-satellite DNA, centromeres, and telomeres, and relative location of chromosomes. RESULT(S): Chromosomes appeared to be oriented inward by centromeres and to be interconnected by major alpha-satellite DNA, which appeared to be the sole DNA component of the nucleoli. This chromosomal arrangement persisted throughout interphase. Chromosomal painting failed to identify chromosomal ordering within pronuclei. CONCLUSION(S): Pronuclear nucleoli are represented by alpha-satellite sequences of interconnecting chromosomes that hold all chromosomes together during interphase. Chromosomes within the pronucleus are randomly positioned relative to each other. PMID- 10685546 TI - Effects of recombinant human FSH (rhFSH), urinary purified FSH (uFSH), and hMG on small preantral follicles and tertiary follicles from normal adult and androgen sterilized female mice. AB - OBJECTIVE: To examine the stage-specific follicular response to recombinant human FSH (rhFSH), urinary FSH (uFSH), and hMG preparations. SETTING: In vitro follicle culture. INTERVENTION(S): Small preantral and tertiary follicles isolated from adult normal BDF-1 mice and androgen-sterilized mice were cultured with rhFSH, uFSH, and hMG for 4 days. MAIN OUTCOME MEASURE(S): Follicular diameter. Immunoreactive inhibin, E2, and progesterone concentrations in cultured medium. RESULT(S): The minimal effective dose of rhFSH, uFSH, and hMG for the follicular growth of small preantral follicles from normal mice was 10 mIU/mL, 1 mIU/mL, and 0.1 mIU/mL, respectively. For tertiary follicles from normal mice, the minimal effective dose of rhFSH, uFSH, and hMG was 10 mIU/mL, 10 mIU/mL, and 1 mIU/mL, respectively. The minimal effective dose of hMG for the follicular growth of small preantral follicles from androgen-sterilized mice was 0.01 mIU/mL, and that of rhFSH and uFSH on tertiary follicles from androgen-sterilized mice was 1 mIU/mL and 10 mIU/mL, respectively. No significant increase was found in the follicular diameter of the tertiary follicles from androgen-sterilized mice as a result of stimulation by hMG, but an hMG dose of >10 mIU/mL produced a significant increase in progesterone secretion. CONCLUSION(S): Human menopausal gonadotropin preparation acts detrimentally on follicles from androgen-sterilized mice by increasing the sensitivity of small preantral follicles to FSH and by inducing the luteinization of tertiary follicles. PMID- 10685547 TI - Temporal analysis of connexin43 protein and gene expression throughout the menstrual cycle in human endometrium. AB - OBJECTIVE: To analyze the pattern of connexin43 gene and protein expression in human endometrium throughout the menstrual cycle. DESIGN: Controlled clinical study. SETTING: An academic research center. PATIENT(S): Women with 28-day menstrual cycles who had mechanical infertility and failed to conceive after IVF treatment. INTERVENTION(S): Endometrial and blood samples were collected on days 8, 12, 14, 21, and 25 of spontaneous menstrual cycles. MAIN OUTCOME MEASURE(S): Endometrial expression of connexin43 protein and messenger RNA, endometrial thickness, and serum concentrations of gonadotropins and steroids. RESULT(S): The expression of connexin43 gene and protein decreased on day 12 and day 14 of the menstrual cycle and then increased on day 21 and day 25, respectively. A serum LH surge accompanied by a peak in the FSH concentration was observed on days 12-14. The progesterone concentration increased on days 21-25, but there was no significant change in the E2 concentration. The thickness of the endometrium increased between days 8 and 12 and did not change further between days 21 and 25. CONCLUSION(S): The expression of connexin43 gene and protein in human endometrium changes during the menstrual cycle in a pattern that is associated with the secretion of LH, FSH, and progesterone. This pattern may serve as a marker for implantation competence. PMID- 10685548 TI - Comparative effects of quinacrine and erythromycin in adult female rats: a nonsurgical sterilization study. AB - OBJECTIVE: To compare the efficacies of erythromycin and quinacrine for nonsurgical sterilization in rats. Quinacrine used for nonsurgical sterilization in women is mutagenic, and most clinical regimens have had a higher failure rate than surgical sterilization. DESIGN: This acute mammal study included five groups of rats assigned randomly and evaluated at two times after treatment. ANIMAL(S): Adult female Sprague-Dawley rats. INTERVENTION(S): Five groups of female Sprague Dawley rats (20 per group) were given 70 or 280 mg/kg of erythromycin lactobionate, 350 mg/kg of quinacrine hydrochloride, or vehicle control administered transcervically. Rats were mated 21 days later. Additional groups (n = 4 per group) were treated and killed 21 days later without mating. MAIN OUTCOME MEASURE(S): Fourteen days after mating, numbers of ovarian corpora lutea, total uterine implants, and embryos were evaluated. For unmated animals, uterine sections were examined for fibrosis and lumen closure. RESULT(S): Neither drug altered numbers of corpora lutea. Erythromycin decreased pregnancy rate and number of implantations (increased preimplantation loss) in a dose-related fashion. Quinacrine increased resorptions. Uterine pathology was more extensive and frequent in erythromycin-treated animals, with extent and severity increasing from 21 to 35+ days. CONCLUSION(S): Erythromycin was more effective than quinacrine in preventing pregnancy. PMID- 10685549 TI - Human sperm microinjection into hamster oocytes: a new tool for training and evaluation of the technical proficiency of intracytoplasmic sperm injection. AB - OBJECTIVE: To design a system for teaching intracytoplasmic sperm injection (ICSI) and to provide a standardized method to assess technical competency. SETTING: University andrology laboratory. DESIGN: Prospective study of method for training ICSI and prediction of ICSI outcome. PATIENT(S): Male infertility candidates for ICSI and fertile donors. INTERVENTION(S): Sperm from 14 fertile donors and 21 oligospermic patients were microinjected into hamster ova. Sperm head decondensation rates (SHD) and oocyte damage rates were measured. Hamster ICSI (HICSI) was used to train technicians, to assess competency, for quality control, and to predict ICSI fertilization. Sperm fertilization potential measured by HICSI was compared with the outcome of ICSI. MAIN OUTCOME MEASURE(S): Sperm head decondensation or fertilization. RESULT(S): Sperm head decondensation was observed in 425 of 773 hamster oocytes with a mean (+/- SD) rate of 60.9 +/- 15.5. Consistency was shown by repetitive testing of the same donor, comparing fresh and frozen semen, and testing of the multiple frozen aliquots of the same ejaculate. Technicians have been trained with this protocol. Excellent initial ICSI success rates for new technicians were demonstrated. Oligospermic semen samples (21 men, 251 hamster ova) tested in the HICSI test exhibited SHD rates from 12% to 100%. The poor outcome of ICSI in clinical cases was predicted by HICSI. CONCLUSION(S): The HICSI provides a method for determining the competency for the ICSI technician and interlaboratory comparison, for the prediction of success of sperm for ICSI. PMID- 10685550 TI - Transcervical placement of a Malecot catheter after hysteroscopic evaluation provides for easier entry into the endometrial cavity for women with histories of difficult intrauterine inseminations and/or embryo transfers: a prospective case series. AB - OBJECTIVE: To evaluate a new technique designed to improve access to the endometrial cavity through tortuous and/or stenotic endocervical canals in women with histories of difficult IUIs, ETs, or endometrial biopsies. DESIGN: Prospective case series. SETTING: Tertiary care center. PATIENT(S): Women with histories of difficult intrauterine procedures because of tortuous and/or stenotic endocervical canals who continued to undergo treatment. INTERVENTION(S): Hysteroscopic evaluation and/or correction of the endocervix, followed by transcervical placement of a Malecot catheter (CR Bard Inc., Covington, GA) for an average of 10 days. MAIN OUTCOME MEASURE(S): Improvement in the ease of access to the endometrial cavity during IUIs or ETs. RESULT(S): Thirty-two of 36 patients had significantly easier procedures after the placement and removal of a Malecot catheter. CONCLUSION(S): Hysteroscopic evaluation and placement of a Malecot catheter is a useful technique that allows easier entry through the cervical canal in patients in whom previous IUIs, ETs, and endometrial biopsies have been difficult. This procedure may lead to improved pregnancy rates, particularly with IVF-ET, as the ease of ET has been correlated with improved implantation rates. PMID- 10685551 TI - Diagnostic accuracy of sonohysterography, transvaginal sonography, and hysterosalpingography in patients with uterine cavity diseases. AB - OBJECTIVE: To evaluate the diagnostic accuracy of sonohysterography (SHG) in uterine cavity diseases in infertile patients, comparing its results with those of hysterosalpingography (HSG) and transvaginal sonography (TVS). Hysteroscopy was the gold standard. DESIGN: Descriptive, prospective study. SETTING: A tertiary university referral center. PATIENT(S): Sixty-five infertile women 19 to 43 years of age. INTERVENTION(S): Patients underwent SHG, conventional TVS, HSG, and hysteroscopy. MAIN OUTCOME MEASURE(S): The results of each examination were compared with those obtained by the gold standard. The following diagnoses were considered separately: polypoid lesions, uterine malformations, intrauterine adhesions, and endometrial hyperplasia (EH). Sensitivity, specificity, positive and negative predictive values (PPV and NPV, respectively), and 95% confidence intervals were calculated. RESULT(S): Sonohysterography had the same diagnostic accuracy as the gold standard for polypoid lesions and EH, with no equivocal diagnosis. Hysterosalpingography showed a sensitivity of 50% and a PPV of 28.6% for polypoid lesions and a sensitivity of 0% for EH. Transvaginal sonography had both sensitivity and PPV of 75% for polypoid lesions and EH. For uterine malformations, SHG had a sensitivity of 77.8%, whereas TVS and HSG both had a sensitivity of 44.4%. Sonohysterography and HSG had a sensitivity of 75% in the detection of intrauterine adhesions and respective PPVs of 42.9% and 50%. Transvaginal sonography showed sensitivity and PPV of 0% for this diagnosis. CONCLUSION(S): Sonohysterography was in general the most accurate test. Its diagnostic accuracy was markedly superior for polypoid lesions and EH, with total agreement with the gold standard. In diagnosis of intrauterine adhesions, SHG had limited accuracy, similar to that obtained by HSG, with a high false-positive diagnosis rate. PMID- 10685552 TI - Predictive value of transvaginal sonography performed before routine diagnostic hysteroscopy for evaluation of infertility. AB - OBJECTIVE: To compare transvaginal sonography with hysteroscopy for the evaluation of intrauterine disorders. DESIGN: Clinical study. SETTING: Academic research environment. PATIENT(S): Patients who were undergoing initial evaluation for primary or secondary infertility or investigation after three failed IVF attempts. INTERVENTION(S): Transvaginal sonography was performed, followed by hysteroscopy, between January 1998 and April 1999. The endometrial findings at sonography were compared with those at hysteroscopy, which served as the gold standard. The characteristic sonographic features of intrauterine adhesions were defined. MAIN OUTCOME MEASURE(S): Intrauterine adhesions, endometrial polyps. RESULT(S): The sensitivity, specificity, and positive and negative predictive values for transvaginal sonography in detecting abnormal uterine cavities were 100%, 96.3%, 91.3%, and 100%, respectively. The corresponding values for the specific diagnoses of intrauterine adhesions and endometrial polyps were 80%, 100%, 100%, and 97%, and 71.4%, 100%, 100%, and 97.1%, respectively. On transvaginal sonography, intrauterine adhesions appeared as hyperechoic endometrial foci and were differentiated from endometrial polyps by their irregular shape and more precise localization. The performance of transvaginal sonography at midcycle (three-layer endometrium) rather than after menstrual cessation (endometrial thickness <3 mm) enabled better imaging of small intrauterine adhesions. CONCLUSION(S): A regular myometrial-endometrial interface and homogeneous endometrial structure on transvaginal sonography congruent with the phase of the menstrual cycle indicated a normal endometrium and precluded the need for diagnostic hysteroscopy. Transvaginal sonography may be used as the initial diagnostic procedure to select patients for hysteroscopy. PMID- 10685553 TI - The need for more CEPHS. PMID- 10685554 TI - The need for more CEPHS. PMID- 10685555 TI - Predictive value of ovarian endometriosis? PMID- 10685556 TI - The value of day 3 follicle-stimulating hormone levels. PMID- 10685557 TI - Concluding letter from Robert F. Harrison on the predictive value of serum FSH levels. PMID- 10685558 TI - Declining human fertility? PMID- 10685559 TI - Craniofacial ballpoint pen injury: endoscopic management. AB - Penetrating facial injuries are not infrequent. There have been isolated case reports of unusual penetrating craniofacial trauma. We describe an unusual case of a 22-month-old child who suffered an external orbital injury from a ballpoint pen that penetrated the orbit, lamina papyracea, posterior ethmoid sinuses, and sphenoid sinus. Endoscopic sinus surgery was performed to extract the ballpoint pen nib after localization with computed tomography. Careful pediatric endoscopic sinus surgery techniques permitted safe foreign body extraction with minimal morbidity. PMID- 10685560 TI - Preliminary clinical results of window partial laryngectomy: a combined endoscopic and open technique. AB - Endoscopic laser resection for anterior commissure glottic carcinoma is difficult, because of inadequate exposure and close proximity to the underlying cartilage. A technique combining endoscopic carbon dioxide laser incision and an external approach creating a window in the thyroid cartilage was initially tested in a canine study and then performed in 5 patients. All patients were men, with T1 or T2 glottic or supraglottic cancer involving the anterior commissure, and had failed radiation treatment. The true or false vocal fold tumors were excised along with the paraglottic space and adjacent cartilage, with preservation of the remaining thyroid framework. The reconstruction was accomplished with placement of a sternohyoid muscle flap, by use of either a bipedicled muscle flap with overlying skin or a unipedicled muscle flap with a graft of free mucosa. The graft was secured in place with fibrin glue and laser soldering. Follow-up ranged from 11 months to 4 years and included biopsies. All patients had voice recordings before and after surgery. A tracheostomy was avoided in all patients. The hospital stays were 4 to 13 days. The voice quality was good after surgery. One patient died of unrelated causes 18 months after his surgery without evidence of recurrence. The other patients are still alive with no evidence of disease. The only complication was subcutaneous neck emphysema in 1 patient that spontaneously resolved. The results showed a satisfactory anatomic reconstruction and acceptable functions. We believe that this new combined technique is oncologically sound, may overcome the limited access seen with the endoscopic technique and the excessive cartilage resection seen with external partial laryngectomy, avoids a tracheostomy, and shortens hospital stays. PMID- 10685561 TI - Quantitative endoscopy: initial accuracy measurements. AB - The geometric optics of an endoscope can be used to determine the absolute size of an object in an endoscopic field without knowing the actual distance from the object. This study explores the accuracy of a technique that estimates absolute object size from endoscopic images. Quantitative endoscopy involves calibrating a rigid endoscope to produce size estimates from 2 images taken with a known traveled distance between the images. The heights of 12 samples, ranging in size from 0.78 to 11.80 mm, were estimated with this calibrated endoscope. Backup distances of 5 mm and 10 mm were used for comparison. The mean percent error for all estimated measurements when compared with the actual object sizes was 1.12%. The mean errors for 5-mm and 10-mm backup distances were 0.76% and 1.65%, respectively. The mean errors for objects <2 mm and > or =2 mm were 0.94% and 1.18%, respectively. Quantitative endoscopy estimates endoscopic image size to within 5% of the actual object size. This method remains promising for quantitatively evaluating object size from endoscopic images. It does not require knowledge of the absolute distance of the endoscope from the object, rather, only the distance traveled by the endoscope between images. PMID- 10685562 TI - Involvement of carbon monoxide in the innervation of the canine cervical esophagus and trachea. AB - We investigated the involvement of carbon monoxide (CO) in the innervation of the canine cervical esophagus and trachea by means of immunohistochemistry using an antiserum against heme oxygenase-2 (HO-2). We detected HO-2-immunoreactive nerve fibers around the blood vessels and submucosal glands of the esophagus and trachea. We found HO-2-immunoreactive neurons in ganglia in the trachea and in the myenteric plexus of the esophagus. These results suggest that CO is involved as a neurotransmitter in the innervation of the canine esophagus and trachea. PMID- 10685563 TI - Comparative histology of the maculae flavae of the vocal folds. AB - A light and electron microscopic comparative investigation of the maculae flavae of the vocal folds was carried out on excised human and canine adult larynges. The structure and functions of human adult maculae flavae (HMF) were found to differ from those of canine adult maculae flavae (CMF). The maculae flavae were composed of fibroblasts, elastic and collagenous fibers, and ground substance in humans and canines. The density of fibroblasts in HMF was found to exceed that in CMF. Fibroblasts in HMF were stellate with processes or spindle-shaped, and the nucleus-cytoplasm (N/C) ratio was small. Rough endoplasmic reticulum and Golgi apparatus were well developed in the cytoplasm. Fibroblasts in CMF were oval, and the N/C ratio was large. Endoplasmic organs were poorly developed in the cytoplasm. Synthesized elastic and collagenous fibers were more numerous in HMF than CMF, and the density of both in HMF was much greater than that in CMF. Ground substance was more abundant in CMF than HMF. Apparently, CMF did not produce elastic and collagenous fibers in amounts sufficient to develop vocal ligaments. The HMF contributes to the formation of the vocal ligaments and the layered structure of human vocal folds. PMID- 10685564 TI - Microsurgery of sulcus vergeture with carbon dioxide laser and injectable collagen. AB - Between January 1989 and June 1998, we operated on 45 patients for sulcus vergeture. The studied population encompassed 38 women (84%) and 7 men (16%). The median age was 36 (range 12 to 71 years). The surgical technique is based on a concept of Cornut and Bouchayer according to which the dissection of the epithelium adherent to the deep subepithelial plane improves the vocal fold vibration. Dissection is performed with a single-pulsed carbon dioxide laser at 2 to 3 W with a pulse duration of 0.1 second. We use the Super-pulse microwave. The Acuspot micromanipulator provides a spot size of 250 microm at 350-mm focal length. When the vocal fold is atrophic, surgery is completed with a bovine or autologous collagen injection; the median injected quantity is 0.3 mL (range 0.1 to 0.4 mL). The epithelial microflap is redraped with fibrin glue. Voice therapy is indispensable for correcting the associated hyperkinetic dysphonia. The median postoperative follow-up period is 5 months (range 1 to 18 months). In terms of median values, the maximum phonation time improved from 9 to 13 seconds, the phonation quotient improved from 296.5 to 228.5 mL/s, and the spectral analysis distribution improved by 1 class. Stroboscopic examination reveals an improvement of the vibratory symmetry, amplitude, and wave. Subjectively, the patients describe an improved ability for vocal effort and the regression or disappearance of vocal fatigue. Although the timbre is improved, the voice often remains breathy and hoarse. PMID- 10685565 TI - Videoendoscopic laryngeal surgery. AB - This paper introduces videoendoscope-assisted laryngeal surgery with office-based equipment. With this technique, a patient is seated and the nose, pharynx, and larynx are topically anesthetized. A flexible videoendoscope with a light sensitive charge-coupled device chip built into the tip is transnasally inserted by an assistant. Specially designed fine-tipped forceps and scalpels were developed for removal of laryngeal lesions. Videoendoscopic laryngeal surgery was undertaken in 114 cases of laryngeal lesions such as polyps, granuloma, and cancer. For benign vocal fold lesions, postoperative vocal function was shown to be improved on aerodynamic and perceptual analyses. For laryngeal tumors, biopsy of the lesion was easily undertaken. Videoendoscopic laryngeal surgery presents the following advantages. It is applicable to outpatients not requiring general anesthesia, it enables functional monitoring of the patient's voice and vocal fold during phonation, it allows for delicate manipulations with both hands, and it gives high-resolution images in comparison to conventional fiberscopy. PMID- 10685566 TI - Curative radiotherapy for anterior commissure laryngeal carcinoma. AB - There is continuing controversy surrounding the most effective treatment of glottic carcinoma involving the anterior commissure (AC). Surgery has been the preferred method of treatment, since studies previously indicated early tumor invasion of the thyroid cartilage at the AC, thereby assuming less curability by radiotherapy (RT). Subsequent laryngeal anatomic studies and refinement of RT techniques have brought into question the ineffectiveness of curative irradiation. A retrospective review of 174 patients with early-stage glottic carcinoma treated with standard fractionation curative RT revealed 34 patients with T1 and T2 lesions involving the AC. Allowing for a follow-up of at least 3 years, we observed only a 12% (4 of 34 patients) local recurrence rate after RT alone, with excellent voice quality and no major complications related to the irradiation. The 4 local recurrences were controlled by total laryngectomy, although 2 patients developed distant metastatic disease. Radiotherapy represents an effective method of treating T1 squamous cell carcinoma of the glottis with AC involvement. The small number of T2 glottic carcinomas in this study prevents a meaningful conclusion concerning treatment of these lesions. PMID- 10685567 TI - Cochlear implantation for auditory rehabilitation in Camurati-Engelmann disease. AB - Camurati-Engelmann disease (progressive hereditary diaphyseal dysplasia) is a rare sclerotic bone disease involving the diaphyses of the long bones, skull base, and clavicles. Progressive sclerosis of cranial nerve foramina has been implicated in cranial nerve deficits. including facial nerve palsy, vestibular disturbances, and hearing loss. Two patients with Camurati-Engelmann disease and concomitant sensorineural hearing loss are presented. Both patients were evaluated for cochlear implantation. One patient was successfully implanted after preoperative imaging revealed no involvement of the internal auditory canals. The porous nature of the affected bone, however. necessitated the inactivation of 1 electrode to prevent facial nerve stimulation. A second patient was rejected as a potential implant recipient due, in part, to narrow internal auditory canals and rapidly progressive disease. The otologic manifestations of Camurati-Engelmann disease are reviewed, and issues related to cochlear implantation in this rare disease are discussed. PMID- 10685568 TI - Experimental study following inactive implantation of an auditory brain stem implant in nonhuman primates. AB - We report changes in the cochlear nuclei (CNs) after 3 months of bilateral auditory deafferentation and simultaneous unilateral implantation of a dummy auditory brain stem implant (ABI) in 6 nonhuman primates (Macaca fascicularis). These specimens were compared to CNs of 9 controls and 7 bilaterally deafferented animals without implantation. The ABI array consists of 3 platinum electrodes mounted on a silicone pad with the back side covered with Dacron. No migration of the ABI was observed. All deafferented animals showed astrocytic reorganization in the CNs. Histologic changes consisted of superficial reactions around the implant, with formation of fibrillar bundles of fusiform cells, and the presence of giant cells close to the Dacron. Other findings were related to surgical trauma. The dummy ABI did not itself provoke serious adverse reactions in the CNs. Our observations support the possibility of ABI reimplantation surgery. PMID- 10685569 TI - Production of diagnostic rules from a neurotologic database with decision trees. AB - A decision tree is an artificial intelligence program that is adaptive and is closely related to a neural network, but can handle missing or nondecisive data in decision-making. Data on patients with Meniere's disease, vestibular schwannoma, traumatic vertigo, sudden deafness, benign paroxysmal positional vertigo, and vestibular neuritis were retrieved from the database of the otoneurologic expert system ONE for the development and testing of the accuracy of decision trees in the diagnostic workup. Decision trees were constructed separately for each disease. The accuracies of the best decision trees were 94%, 95%, 99%, 99%, 100%, and 100% for the respective diseases. The most important questions concerned the presence of vertigo, hearing loss, and tinnitus; duration of vertigo; frequency of vertigo attacks; severity of rotational vertigo; onset and type of hearing loss; and occurrence of head injury in relation to the timing of onset of vertigo. Meniere's disease was the most difficult to classify correctly. The validity and structure of the decision trees are easily comprehended and can be used outside the expert system. PMID- 10685570 TI - Rheumatoid arthritis of the temporomandibular joint with herniation into the external auditory canal. AB - Previous authors have shown that soft tissue can present in the external auditory canal via a patent foramen of Huschke. One case represented a patient with psoriatic arthritis and a polyp in the external auditory canal. Typically, neoplastic, inflammatory, or degenerative lesions of the temporomandibular joint do not present in the external auditory canal. We present a patient with rheumatoid arthritis of the temporomandibular joint and soft tissue herniation into the external auditory canal. The case, and a discussion of possible causes, are presented. PMID- 10685571 TI - Immunohistochemical localization of collagen types I, III, and IV, laminin, fibronectin, and keratin in the endolymphatic sac. AB - We have employed immunohistochemistry to obtain baseline information on the molecular constituents of the extracellular matrix (ECM) of the endolymphatic duct (ED) and endolymphatic sac (ES) of the chinchilla. The results demonstrated that collagen types I and III were distributed in the subepithelial layer in the ED and ES, type IV collagen and laminin in the basement membranes, and fibronectin in the subepithelial layer and partly in the conglomerated cells in the ES. Collagen type III was diffusely distributed in the whole subepithelial layer of the ES, whereas collagen type I was concentrated densely in the deep layer of the interstitium, although gradually, the cuboidal epithelium in the ES was transformed into a flatter type in the ED. The epithelial cells of the ED and ES were clearly positive for keratin. This study deals, in particular, with the normal distribution of ECM components of the ED and ES of the chinchilla. PMID- 10685572 TI - Midline lateralization thyroplasty for adductor spasmodic dysphonia. AB - Midline lateralization thyroplasty was successfully performed on a patient with adductor spasmodic dysphonia. The thyroid cartilage was incised at the midline, and a 3 x 2-mm perforation was made at the anterior commissure to widen it. The perforation was closed with a free composite graft taken from the upper edge of the thyroid ala, and the incised thyroid cartilage edges were kept separated 4 mm apart with silicone wedges. A part of the sternohyoid muscle was rotated to seal any leak from the perforation. The postoperative course was uneventful. The voice has been restored to normal, and there is no sign of recurrence of the symptom so far, as of 1 year 5 months postoperative. Although a longer follow-up is needed, this case indicates that midline type II thyroplasty could be a useful treatment for adductor spasmodic dysphonia. PMID- 10685573 TI - Effects of botulinum toxin on pathophysiology in spasmodic dysphonia. AB - To determine the mechanism of symptom relief with treatment by botulinum toxin injection in persons with adductor spasmodic dysphonia (ADSD), we evaluated the effects of unilateral thyroarytenoid muscle injections on both injected and noninjected muscles in 10 subjects with ADSD, using electromyography on both sides of the larynx before and after treatment. The subjects' speech symptoms were reduced (p = .005) 2 weeks following injection, when the electromyographic study occurred. Muscle activation levels and the numbers of spasmodic muscle bursts decreased significantly (p < or = .03) postinjection in both the injected and noninjected muscles. The reductions in laryngeal muscle bursts correlated with symptom reduction (r > or = .7) in all muscles. Reductions in laryngeal muscle bursts did not relate to either absolute or normalized levels of muscle activity before or after botulinum toxin injection. The results suggest that changes in the central pathophysiology are responsible for changes in speech symptoms following treatment. PMID- 10685574 TI - Botulinum toxin injections for essential voice tremor. AB - Fifteen patients, 13 women and 2 men, with a mean age of 72.7 years (56 to 86 years) and a clinical diagnosis of essential voice tremor, were treated with botulinum injections to the thyroarytenoid muscles, and in some cases, to the cricothyroid or thyrohyoid muscles. Evaluations were based on subjective judgments by the patients, and on perceptual and acoustic analysis of voice recordings. Subjective evaluations indicated that the treatment had a beneficial effect in 67% of the patients. Perceptual evaluations showed a significant decrease in voice tremor during connected speech (p < .05). Acoustic analysis showed a nearly significant decrease in the fundamental frequency variations (p = .06) and a significant decrease in fundamental frequency during sustained vowel phonation (p < .01 ). The results of perceptual evaluation coincided most closely with the subjective judgments. It was concluded that the treatment was successful in 50% to 65% of the patients, depending on the method of evaluation. PMID- 10685575 TI - Mucociliary transport pathway on laryngotracheal tract and stented glottis in guinea pigs. AB - We investigated the laryngotracheal mucociliary transport pathway of guinea pigs in vivo and immediately postmortem. Only intraperitoneal anesthesia was used during the procedure to avoid the disturbance of mucociliary function. Resin particles were used as the marking substance. A microcolpohysteroscope was placed at different levels in the laryngotracheal region for observing the marking particles and recording the transport pattern. The tracheal mucociliary transport flow primarily moved along the posterior wall and both lateral walls in a zigzag trace. Upon reaching the subglottis, the resin particles stayed underneath the vocal cords, and a whirlpool phenomenon developed. The majority of the particles were shifted and directed onto the posterior glottic area. With a short delay, some resin particles crossed over the free edge of the vocal cords and turned posteriorly along the medial upper cordal margin. No mucociliary transport could be observed on the entire upper surface of the true vocal cords, which is covered by squamous epithelium. Occasionally, a few resin particles in the vicinity of the epiglottic root traveled along the aryepiglottic folds toward the posterior commissure. All streams of mucociliary transport finally joined together in the interarytenoid area. After leaving the glottis, the resin particles traveled to the hypopharynx and entered the esophagus through the motion of deglutition. The pattern of mucociliary clearance in the laryngotracheal region was not delayed by stenting. PMID- 10685576 TI - Myosin heavy chain expression in human laryngeal muscle fibers. A biochemical study. AB - Since the intrinsic laryngeal muscles in humans are involved in specialized functions, one may suppose that this would be associated with the expression of specific myosin heavy chain (MHC) isoforms, as has been reported for the rat, dog, and rabbit. In order to determine which MHCs are expressed in the human laryngeal muscles, biochemical analysis using sodium dodecyl sulfate polyacrylamide gel electrophoresis was performed. Thyroarytenoid and posterior cricoarytenoid muscles were obtained from a 7-month-old infant and 4 adults. In the adult human laryngeal muscles, 3 bands were resolved identical to those previously described in the human limb muscles (I, IIA, and IIB MHCs). In contrast, muscles from the infant also expressed fetal MHC and a novel MHC not observed in other human skeletal muscles. This novel band migrated at the same level as the laryngeal MHC previously described in the rat. Since these 2 isoforms disappear in the adult, the persistence in the infant could be correlated with the immature development of laryngeal functions and, in particular, phonation. PMID- 10685577 TI - Histopathologic study of alternative substances for vocal fold medialization. AB - This research investigated the histopathologic and migratory properties of injectable alternatives for vocal fold medialization. Thirteen dogs underwent sectioning of the recurrent laryngeal nerve followed by vocal fold injection with 1 of 4 substances: Teflon, autologous fat, silicone suspension, or hydroxyapatite cement. Six months later, the animals were painlessly sacrificed and histopathologic analysis of the larynx and regional lymph nodes was performed. Although regional lymph node migration was noted, Teflon injection resulted in minimal vocal fold inflammatory reaction. Vocal folds injected with autologous fat exhibited persistence of fat at the injection site without significant inflammation or migration. Silicone suspension caused a localized giant cell reaction without regional lymph node migration, and 1 study subject died secondary to acute inflammation with critical respiratory compromise. Hydroxyapatite cement was well tolerated without inflammation or migration. This pilot study indicates that a wide range of possible substances for vocal fold medialization exist. Many of these may produce results superior to those obtained with Teflon and are thus far untested. PMID- 10685578 TI - Sinusitis and carotid artery stroke. AB - The relationship between sinusitis and ischemic stroke is unexplored. The anatomic proximity between the paranasal sinuses and the internal carotid artery suggests that inflammation of the sinuses could easily extend to the intracranial vasculature. We report 4 patients with acute ischemic stroke and extensive disease of the paranasal sinuses. All patients had large vessel stroke involving the internal carotid artery territory. All patients had extensive disease of the sphenoid and other sinuses. The sinus disease was demonstrated by magnetic resonance imaging. These case report observations suggest a relationship between inflammation of the paranasal sinuses, particularly sphenoid sinusitis, and ischemic stroke. PMID- 10685579 TI - Aneurysm of an aberrant right subclavian artery presenting as dysphagia lusoria. PMID- 10685580 TI - Hyalinizing trabecular adenoma of the thyroid diagnosed by fine-needle aspiration biopsy. PMID- 10685581 TI - A trial to determine the risk of decompression sickness after a 40 feet of sea water for 200 minute no-stop air dive. AB - BACKGROUND: The USN93 probabilistic model of decompression sickness (DCS) predicts a DCS risk of 3.9% after a 40 ft of seawater (fsw) for 200 min no-stop air dive, although little data is available to evaluate the accuracy of this prediction. Based on an analysis of Navy Safety Center data from diving on U.S. Navy standard air decompression tables, the observed incidence of DCS for this type of dive is 0.11%. Knowing the true incidence of the dive is important for deciding whether or not to adopt proposed probability based decompression procedures for U.S. Navy diving. HYPOTHESIS: The risk of DCS after a 40 fsw for 200 min no-stop air dive is 3.9%. METHODS: We conducted a closed sequential trial to determine the DCS incidence on this dive. RESULTS: Of 30 military divers who completed 91 dives, there were 2 cases of DCS (2.2%, 95% CI: 0.27 7.7%). The study was terminated early after the second DCS case because of the presence of neurological symptoms and signs. CONCLUSIONS: This study demonstrates that the incidence of DCS in a laboratory setting is higher than observed in fleet diving. Use of the 40 fsw for 200 min schedule in a decompression computer is likely to result in DCS incidence 2.5- to 70-fold greater than that observed in U.S. Navy diving using table-based procedures. PMID- 10685582 TI - Escape from a disabled submarine: decompression sickness risk estimation. AB - Individual crewmember escape from a disabled U.S. Navy nuclear submarine has never been necessary, but remains an important contingency. Decompression sickness (DCS) is one of the foreseeable risks and a robust mathematical model of DCS incidence has been used to estimate the magnitude of this risk under a variety of escape scenarios. The model was calibrated with over 3000 well controlled human pressure exposures, less than 2% of which simulated pressure profiles of submarine escape. For disabled submarine depths < 300 ft of sea water (fsw) and internal submarine pressures of <11 fsw (arguably the most likely conditions), the DCS risks are comparable to those routinely undertaken by U.S. Navy divers--less than 5%. For progressively deeper depths and especially for higher submarine internal pressures, the risk of DCS becomes much greater, including unknown chances of permanent injury and death. Variations from the baseline escape procedure are explored, including equipment differences, delays in exiting the submarine and changes in the oxygen content of the breathing mix. PMID- 10685583 TI - The effectiveness of ground level oxygen treatment for altitude decompression sickness in human research subjects. AB - BACKGROUND: Current therapy for altitude decompression sickness (DCS) includes hyperbaric oxygen therapy and ground-level oxygen (GLO). The purpose of this paper is to describe the Air Force Research Laboratory experience in the extensive use of GLO for the treatment of altitude DCS in research subjects. METHODS: Data were collected from 2001 altitude chamber subject-exposures. These data, describing DCS symptoms, circulating intracardiac venous gas emboli, and treatment procedures used were collected for each subject exposure and stored in an altitude DCS database. RESULTS: In the database of 2001 subject exposures, 801 subjects (40.0%) were diagnosed with altitude DCS. Subjects reporting DCS symptoms were immediately recompressed to ground level. Of the 749 subjects who received 2 h GLO, 739 (98.7%) resolved completely and required no further treatment. CONCLUSIONS: Although not an operational study, these data provide indirect support for the current USAF guidelines for the treatment of altitude DCS with GLO. PMID- 10685584 TI - Complications and side effects of hyperbaric oxygen therapy. AB - BACKGROUND: Despite ongoing controversy, hyperbaric oxygen (HBO) therapy is frequently administered in various clinical situations. Probably because of the unique atmospheric conditions to which the patient is exposed, there are concerns about the safety aspects of this therapy. Possible complications during HBO therapy include barotraumatic lesions (middle ear, nasal sinuses, inner ear, lung, teeth), oxygen toxicity (central nervous system, lung), confinement anxiety, and ocular effects (myopia, cataract growth). METHODS: To analyze the medical safety of HBO therapy, this report reviewed complications and side effects of 782 patients treated for various indications with a total of 11,376 HBO therapy sessions within a multiplace chamber. The absolute treatment pressure was 240 or 250 kPa 114 or 15 msw). The compression was performed in a linear manner with 14 to 15 kPa (1.4 to 1.5 msw) x min(-1). All data were gathered prospectively within a special database. RESULTS: More than 17% of all patients experienced ear pain or discomfort as an expression of problems in equalizing the middle ear pressure. Most episodes were not related to a persistent eustachian tube dysfunction since they only occurred once. Barotraumatic lesions on visual otological examinations (ear microscopy) were verified in 3.8% of all patients. Patients with sensory deficits involving the ear region need special attention, because they seem to be at risk for rupture of the tympanic membrane (three cases documented). A barotrauma of the nasal sinuses occurred rarely and no barotraumatic lesions of the inner ear, lung, or teeth were noted. Oxygen toxicity of the CNS manifested by generalized seizures affected four patients without any recognizable risk factors or prodromes. None of the patients suffered recurrences or sequelae. Regular checks of the blood glucose in diabetics failed to reveal episodes of hypoglycemia as a cause for seizures. Lung function tests of patients undergoing prolonged treatment (average 52.8 sessions) did not deteriorate. CONCLUSION: Patients scheduled for HBO therapy need a careful pre examination and monitoring. If safety guidelines are strictly followed, HBO therapy is a modality with an acceptable rate of complications. The predominant complication is represented by pressure equalization problems within the middle ear. Serious complications rarely occur. PMID- 10685585 TI - Intermittent hypobaric hypoxia induces altitude acclimation and improves the lactate threshold. AB - The physiological responses to short-term intermittent exposure to hypoxia in a hypobaric chamber were evaluated. The exposure to hypoxia was compatible with normal daily activity. The ability of the hypoxia program to induce hematological and ventilatory adaptations leading to altitude acclimation and to improve physical performance capacity was tested. Six members of a high-altitude expedition were exposed to intermittent hypoxia and low-intensity exercise (in cycle-ergometer) in the INEFC-UB hypobaric chamber over 17 d, 3-5 h x d(-1), at simulated altitude of 4,000 m to 5,500 m. Following this hypoxia exposure program, significant increases were found in packed cell volume (41 to 44.6%; p<0.05), red blood cells count (4.607 to 4.968 10(6) cells x microL(-1); p<0.05), and hemoglobin concentration (14.8 to 16.4 g x dL(-1); p<0.05), thus implying an increase in the blood oxygen transport capacity. Significant differences in exercise blood lactate kinetics and heart rate were also observed. The lactate vs. exercise load curve shifted to the right and heart rate decreased, thus indicating an improvement of aerobic endurance. These results were associated with a significant increase in the ventilatory anaerobic threshold (p<0.05). Significant increases (p<0.05) in pulmonary ventilation, tidal volume, respiratory frequency, O2 uptake, CO2 output and ventilatory equivalents to oxygen (VE/Vo2) and carbon dioxide (VE/co2) were observed at the ventilatory threshold and within the transitional zone of the curves. We conclude that short term intermittent exposure to moderate hypoxia, in combination with low-intensity exercise in a hypobaric chamber, is sufficient to improve aerobic capacity and to induce altitude acclimation. PMID- 10685586 TI - Urinary continence in women during centrifuge exposure to high +Gz. AB - BACKGROUND: One earlier study and anecdotal evidence suggest a possible association between exposure to high +Gz forces and urinary incontinence in women. High +Gz could possibly contributes to the prolapse of the bladder neck, moving it into a position which decreases the leak point pressure resulting in urinary incontinence. HYPOTHESIS: We tested the hypothesis that increased urinary incontinence is associated with high +Gz. METHODS: 25 females were exposed to a high +Gz profile. Following the exposure they were asked to answer a questionnaire grading their urinary continence under high +Gz, and to provide a baseline grading of their urinary continence at +1.0 Gz and under increased abdominal stress at +1.0 Gz. Demographic data included parity and previous urogenital surgery. Graded responses were dichotomized and data was analyzed using Fischer's Exact Test for 2x2 tables with significance set at alpha = 0.05. RESULTS: At high +Gz no significant association was found between reported urine incontinence and a history of urogenital surgery or parity. Only one of twenty five subjects had any symptoms at high +Gz despite the fact that five had a predisposition. As expected, at +1.0 Gz and under increased abdominal stress at +1.0 Gz a significant association was found between reported urine incontinence and a history of urogenital surgery, while no significant association was found for parity. CONCLUSIONS: In this simple first look there was no increase in urinary incontinence at high +Gz even among those who reported a predisposition. PMID- 10685587 TI - Heart rate and blood pressure responses to +Gz following varied-duration -Gz. AB - BACKGROUND: The push-pull effect has been defined previously as decreased +Gz tolerance caused by previous baseline zero or -Gz exposure. Earlier work indicates that the delay in BP (BP) recovery during +Gz is a function of time at G7, and is due to the lengthened time-course of sympathetically mediated peripheral vasoconstriction. HYPOTHESIS: The purpose of this study was to retrospectively determine whether heart rate (HR) varies with BP as duration at preceding -Gz increased. METHODS: Continuous ECG R-R interval data from 15 s of +2.25Gz after preceding 2, 5, 10, or 15 s at 2Gz obtained from previous experiments were analyzed and compared with the previously reported BP data. Repeated measures ANOVA and regression analyses were used to compare +2.25Gz HR responses after the four -Gz conditions and one control +2.25Gz condition. RESULTS: An initial rapid rise in HR was observed for all conditions with a consistent steady-state plateau achieved after the first 7 s of +2.25Gz. However, there were significant differences in mean HR attained during the +2.25Gz plateau for preceding 15 s -2.0 Gz vs. the control, 2, 5, and 10s -Gz conditions (109+/ 1.1 vs. 102+/-1.8, 100+/-2.0, 97+/-1.1 and 101+/-1.1, bpm, respectively; p<0.05). CONCLUSIONS: HR, unlike BP, increases briskly across all preceding -Gz time conditions, adapting within the initial baroreflex-compensatory time frame typically expected for +Gz exposures. These results suggest there may be a threshold effect for HR response. Consequently, vasoconstrictor response is a critical adaptive mechanism during +Gz when preceded by long (>10 s) -Gz exposures. PMID- 10685588 TI - Vasomotor sympathetic nerve activity in men during bed rest and on orthostasis after bed rest. AB - BACKGROUND: Alterations in autonomic function are commonly seen during and after spaceflight, and its ground-based analog, 6 degrees head-down bed rest (HDBR). They may include peripheral vascular regulation, but vasomotor sympathetic efferent nerve discharges to peripheral vasculatures have not been examined. The aim of our study was to examine changes in vasomotor sympathetic nerve activity during HDBR and under orthostasis after HDBR. METHODS: We performed 6 d of HDBR on six male subjects, and measured muscle sympathetic nerve activity (MSNA) together with plasma norepinephrine concentrations in the supine position before HDBR and in 6 degrees head-down position on the sixth day (HDBR6) of HDBR. We also measured MSNA in head-up tilt (HUT) test before and after HDBR. RESULTS: On HDBR6, MSNA burst rate was the same (17+/-4 bursts x min(-1)) as that in supine position before HDBR (15+/-2 bursts min(-1)), but plasma norepinephrine concentrations were decreased to 1.14+/-0.10 pmol x ml(-1) compared with the supine value before HDBR (1.56+/-0.20 pmol x ml(-1), p<0.05). After HDBR, supine MSNA burst rate significantly increased by 58% to 24+/-4 bursts x min(-1). MSNA increment in response to HUT was similar between before (34+/-3 bursts min(-1) x sin HUT(-1)) and after (40+/-6 bursts x min(-1) x sin HUT(-1)) HDBR. CONCLUSIONS: Our findings suggest that: a) the relationship between MSNA and plasma norepinephrine concentrations was altered on the sixth day during HDBR; b) the vasomotor sympathetic nerve activity was enhanced after HDBR; and c) the augmentation of vasomotor sympathetic outflow to muscles under orthostasis was preserved after HDBR. PMID- 10685589 TI - Exercise thermoregulation in men after 1 and 24-hours of 6 degrees head-down tilt. AB - BACKGROUND: Exercise thermoregulation is dependent on heat loss by increased skin blood flow (convective and conductive heat loss) and through enhanced sweating (evaporative heat loss). Reduction of plasma volume (PV), increased plasma osmolality, physical deconditioning, and duration of exposure to simulated and actual microgravity reduces the ability to thermoregulate during exercise. HYPOTHESIS: We hypothesized that 24 h of head down tilt (HDT24) would alter thermoregulatory responses to a submaximal exercise test and result in a higher exercise rectal temperature (Tre) when compared with exercise Tre after 1 h of head down tilt (HDT1). METHODS: Seven men (31+/-SD 6 yr, peak oxygen uptake (VpO2peak) of 44+/-6 ml x kg(-1) x min(-1)) were studied during 70 min of supine cycling at 58+/-SE 1.5% VO2peak at 22.0 degrees C Tdb and 47% rh. RESULTS: Relative to pre-tilt sitting chair rest data, HDT1 resulted in a 6.1+/-0.9% increase and HDT24 in a 4.3+/-2.3% decrease in PV (delta = 10.4% between experiments, p<0.05) while plasma osmolality remained unchanged (NS). Pre exercise Tre was elevated after HDT24 (36.71 degrees C +/-0.06 HDT1 vs. 36.93 degrees C+/-0.11 HDT24, p<0.05). The 70 min of exercise did not alter this relationship (p<0.05) with respective end exercise increases in Tre to 38.01 degrees C and 38.26 degrees C (degrees = 1.30 degrees C (HDT1) and 1.33 degrees C (HDT24)). While there were no pre-exercise differences in mean skin temperature (Tsk), a significant (p<0.05) time x treatment interaction occurred during exercise: after min 30 in HDT24 the Tsk leveled off at 31.1 degrees C, while it continued to increase reaching 31.5 degrees C at min 70 in HDT1. A similar response (NS) occurred in skin blood velocity. Neither local sweating rates nor changes in body weight during exercise of -1.63+/-0.24 kg (HDT1) or - 1.33+/-0.09 kg (HDT24) were different (NS) between experiments. CONCLUSION: While HDT24 resulted in elevated pre-exercise Tre, reduced PV, attenuation of Tsk and skin blood velocity during exercise, the absolute increase in exercise Tre was not altered. But if sweat rate and cutaneous vascular responses were similar at different core temperatures (unchanged thermoregulation), the Tre offset could have been caused by the HDT-induced hypovolemia. PMID- 10685590 TI - Stretch- and H-reflexes of the lower leg during whole body cooling and local warming. AB - BACKGROUND: This study was undertaken to evaluate if possible changes in stretch- and H-reflexes could be related to the changes in the EMG activity of the cooled lower leg muscles observed during a stretch-shortening cycle exercise. METHODS: Eight subjects wearing shorts and jogging shoes were exposed once to 27 degrees C and twice to 10 degrees C for 60 min each. During the second exposure to 10 degrees C, the subject's lower legs were kept warm (10 degrees Clw) with electrical pillows. After the exposures Achilles tendon reflex (stretch reflex) was induced and the EMG activity of the triceps surae was measured. Immediately after reflex measurements the EMG activity of the triceps surae and tibialis anterior during a drop-jump (stretch-shortening cycle) was measured. After similar thermal exposures electrically induced H-reflex from the calf was measured. RESULTS: During the preactivity and stretch phases the EMG activity of the triceps surae increased after the exposure to 10 degrees C, whereas during the shortening phase it decreased. During the shortening phase cooling, on the contrary, increased the activity of tibialis surae anterior. These changes disappeared at 10 degrees Clw. At 10 degrees C the maximum EMG-amplitude of triceps surae during stretch reflex decreased (p<0.05), reflecting suppressed muscle spindle activity. Suppressed spindle activity causes the agonist to be unfacilitated and the antagonist muscle contraction to be uninhibited, which was seen in the present study as decreased agonist and increased antagonist EMG activity during the shortening phase at 10 degrees C. The Hmax/Mmax-ratio, H reflex latency and amplitude increased at 10 degrees C (p<0.05), reflecting increased motoneuron pool excitability. This in part may explain the increased EMG activity during the preactivity and stretch phases. CONCLUSION: Cooling induced increase in the excitability of the motoneuron pool and suppression of muscle spindle activity seem to be responsible of the EMG activity changes during the stretch-shortening cycle, consequently decreasing muscular performance. PMID- 10685591 TI - Improving athletic performance: is altitude residence or altitude training helpful? AB - Exercise training studies conducted at different altitudes (1250-5700 m) of varying durations (30 min to 19 wk) are critically reviewed to determine the efficacy of using altitude as a training stimulus to enhance sea level and altitude exercise performance. Four strategies are discussed: a) exercise training while residing at the same altitude; b) exercise training at altitude but residing at sea level; c) exercise training at low altitude but residing at a higher altitude; and d) exercise training under sea level and altitude conditions but only after altitude acclimatization has occurred. Residing at altitude causes a multitude of potentially beneficial physiological, ventilatory, hematological and metabolic changes that theoretically should induce a potentiating effect on endurance exercise performance. While it is accepted that endurance performance is greatly enhanced at altitude, there is less support for the view that altitude training while residing at altitude improves subsequent sea level endurance performance. There is some evidence, though also not universally accepted, that training at altitude but residing at sea level may benefit sea level endurance performance. Most recently, the combination of "living high" (e.g., at 2500 m) to obtain beneficial physiological changes associated with altitude acclimatization and "training low" (e.g., at 1250 m) to allow maintenance of high-intensity training is accumulating scientific and popular support as the most advantageous strategy to improve subsequent sea level exercise performance in well-trained, competitive runners. PMID- 10685592 TI - Pulmonary cyst and cerebral arterial gas embolism in a hypobaric chamber: a case report. AB - This is a report of an aircrew member who suffered a serious physiological incident in the form of pulmonary barotrauma and cerebral arterial gas embolism during hypobaric chamber training, and who subsequently was shown to have a cyst in the upper lobe of the left lung. The likely origin of the cyst is discussed, as well as the aeromedical disposition following thoracotomy and apical segmentectomy to remove the cyst. PMID- 10685593 TI - In-flight cerebral oxygen status: continuous monitoring by near-infrared spectroscopy. AB - BACKGROUND: Little is known about the in-flight cerebral oxygen status (COS) of fighter pilots. The purpose of this study was to evaluate the COS of fighter pilots during an F-15DJ flight. METHODS: Three male F-15DJ fighter pilots volunteered to serve as test subjects during aerial gunnery training (AGT) missions. During the flight, the pilots' COS was continuously monitored from the right forehead using a NIRO-300G near-infrared spectrophotometer. This new instrument is capable of measuring the concentration changes in the brain of oxygenated hemoglobin (O2Hb), reduced hemoglobin (HHb), total hemoglobin (cHb, O2Hb + HHb), and the tissue oxygenation index (TOI, O2Hb/cHb). RESULTS: Continuous changes of COS within the brain were clearly demonstrated using NIRO 300G, the changes included reductions in O2Hb, cHb, and TOI with increased +Gz, in a mirror image fashion. The maximum decreased concentration of hemoglobin during flight ranged from 12.8 to 25.6 micromol x L(-1) (of brain tissue). CONCLUSIONS: We believe this study is the first monitoring of COS during F-15 actual flight. The monitoring of COS during F-15 flight showed that the effect of +Gz was to simultaneously lower blood flow in a mirror image fashion. Our results demonstrated that near-infrared spectroscopy (NIRO-300G) provides a reliable and sensitive method for the monitoring of pilots' COS during flight. PMID- 10685594 TI - Physiological problems associated with wearing NBC protective clothing during cold weather. AB - This report considers how thermal balance of soldiers wearing nuclear, biological and chemical (NBC) protective clothing in combination with the Extreme Cold Weather Clothing System (ECWCS) is affected during work in cold weather. A review of published reports concerning physiological consequences of wearing NBC protective clothing during cold exposure was completed. The findings reported in the experimental literature were too limited to adequately forecast the effects of adding NBC clothing to ECWCS. To remedy the information gap, simulation modeling was employed to predict body temperature changes during alternating bouts of exercise and rest throughout 8 h of exposure to three different severely cold conditions. Published findings indicate that NBC protective clothing may inadequately protect against hand and finger cooling, especially during rest following strenuous activity. No evidence substantiates suggestions that wearing NBC protective masks increases susceptibility to facial frostbite. Collectively, the limited experimental work and the results of simulation modeling argue against any increased risk of hypothermia associated with wearing NBC protective clothing while working in the cold. However, wearing NBC protective clothing during strenuous activity in cold weather may increase the risk of hyperthermia, and cause sweat accumulation in clothing which may compromise insulation and increase the risk of hypothermia during subsequent periods of inactivity. PMID- 10685595 TI - Complementary medicine and aviation medicine. PMID- 10685596 TI - NASA opens new virtual airport control tower at Ames. PMID- 10685597 TI - Reports propose ways to better protect military troops from exposure to hazardous agents. PMID- 10685598 TI - Tubulin-tyrosine ligase, a long-lasting enigma. AB - Tubulins and microtubules are subjected to several post-translational modifications of which the reversible detyrosination/tyrosination of the carboxy terminal end of most alpha-tubulins has been extensively analysed. This modification cycle involves a specific carboxypeptidase and the activity of the tubulin-tyrosine ligase (TTL). The true physiological function of TTL has so far not been established. This review describes the purification of TTL to homogeneity by biochemical methods, its in vitro properties and the generation of monoclonal antibodies. These mabs not only enabled a very convenient and rapid purification of TTL by immunoaffinity chromatography but also its extensive characterization by protein sequencing, which led to the isolation of the full length cDNA. With this information, gene disruption should be feasible in order to determine the physiological significance of the tyrosination cycle. PMID- 10685599 TI - The third tubulin pool. AB - Tubulin normally undergoes a cycle of detyrosination/tyrosination on the carboxy terminus of its alpha-subunit and this results in subpopulations of tyrosinated tubulin and detyrosinated tubulin. Brain tubulin preparations also contain a third major tubulin subpopulation which is non-tyrosinatable. This review describes the purification and the structural characterization of non tyrosinatable tubulin. This tubulin variant lacks a carboxyterminal glutamyl tyrosine group on its alpha-subunit (delta2-tubulin). Delta2-tubulin is generated from detyrosinated tubulin through an irreversible reaction. Delta2-tubulin accumulates in neurons and in stable microtubule assemblies. It also accumulates in some tumor cells due to the frequent loss of tubulin tyrosine ligase in such cells. Delta2-tubulin may be a useful marker of malignancy in human tumors. PMID- 10685600 TI - Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl m-toluamide. AB - Pyridostigmine bromide (PB), an inhibitor of acetylcholinesterase, has been used as a prophylactic for nerve gas poisoning. N,N'-diethyl-m-toluamide (DEET) is the active ingredient in most insect repellents and is thought to interact synergistically with PB. Since PB can inhibit the binding of organophosphates to tubulin and since organophosphates inhibit microtubule assembly, we decided to examine the effects of PB and DEET on microtubule assembly as well as their interactions with tubulin, the subunit protein of microtubules. We found that PB binds to tubulin with an apparent Kd of about 60 microM. PB also inhibits microtubule assembly in vitro, although at higher concentrations PB induces formation of tubulin aggregates of high absorbance. Like PB, DEET is a weak inhibitor of microtubule assembly and also induces formation of tubulin aggregates. Many tubulin ligands stabilize the conformation of tubulin as measured by exposure of sulfhydryl groups and hydrophobic areas and stabilization of colchicine binding. PB appears to have very little effect on tubulin conformation, and DEET appears to have no effect. Neither compound interferes with colchicine binding to tubulin. Our results raise the possibility that PB and DEET may exert some of their effects in vivo by interfering with microtubule assembly or function, although high intracellular levels of these compounds would be required. PMID- 10685601 TI - Tubulin carboxypeptidase/microtubules association can be detected in the distal region of neural processes. AB - The association of tubulin carboxypeptidase with microtubules has been demonstrated in crude brain extracts and in living non-nervous cells. Here, we studied this phenomenon in cultured brain cells. To determine the association of the enzyme with neural microtubules we isolated the cytoskeletons (detergent extraction under microtubule-stabilizing conditions) and measured the content of Tyr, Glu, and delta2 tubulin as a function of the in vitro incubation time of the cytoskeletons. The carboxypeptidase was found associated with microtubules in 2 days-cultured cells but not in 7 days-cultured cells. Quantitative analysis of digitized images after immunofluorescent staining revealed that detyrosination during the incubation of the cytoskeletons occurred preferentially in the distal regions of the neural processes. Prolonged taxol-treatment of the cells promoted higher detyrosination but Tyr tubulin was not depleted suggesting the existence of a subset of microtubules that has not associated carboxypeptidase and therefore cannot be detyrosinated even after prolonged taxol-treatment. This hypothesis was supported, although not conclusively, by additional experiments. PMID- 10685602 TI - Tau protein function in axonal formation. AB - Tau protein is a predominantly neuronal microtubule-associated protein that is enriched in axons and is capable of promoting microtubule assembly and stabilization. In the present article we review some of the key experiments directed to obtain insights about tau protein function in developing neurons. Aspects related to whether or not tau has essential, unique, or complementary functions during axonal formation are discussed. PMID- 10685603 TI - Tau dephosphorylation at tau-1 site correlates with its association to cell membrane. AB - It has been considered that tau protein is mainly a cytoplasmic protein since it is a microtubule associated protein. However, it has also been suggested that tau could be located in the cell nucleus and membrane. In our work, the cellular distribution of tau has been studied by immunofluorescence and western blot analysis, after subcellular fractionation in neuroblastoma cells and in tau transfected non neural cells using, mainly, two types of tau antibodies; antibody 7.51 (that recognizes tau independent of its phosphorylation level); and antibody Tau-1 (that recognizes tau only in its dephosphorylated form). Also, tau was expressed in COS-1 cells to test for the features involved in the sorting of tau to different cell localizations. Our results show that tau associated to cell membrane has a lower phosphorylation level in its proline-rich region. Additionally, in differentiated neuroblastoma cells, tau phosphorylation, at that region, decreases and the amount of tau associated to cell membrane increases. PMID- 10685604 TI - What is the signal for the posttranslational arginylation of proteins? AB - The N-terminal, posttranslational arginylation of proteins is ubiquitous in eukaryotic cells. Previous experiments, using purified components of the reaction incubated in the presence of exogenous substrates, have shown that only those proteins containing acidic residues at their N-terminals are arginylation substrates. However, data from experiments that used crude extracts of brain and nerve as the source of the arginylating molecules, suggest that the in vivo targets for arginylation are more complex than those demonstrated using purified components. One of the proposed functions for arginylation is as a signal for protein degradation and proteins that have undergone oxidative damage have been shown to be rapidly degraded. In the present experiments we have tested the hypothesis that the presence of an oxidatively damaged residue in a protein is a signal for its arginylation. These experiments have been performed by adding synthetic oxidized peptides to crude extracts of rat brain, incubating them with [3H]Arg and ATP and assaying for arginylated peptides using RP-HPLC. Results showed that while the oxidized A-chain of insulin was arginylated in this system, confirming previous experiments, other peptides containing oxidized residues were not. When a peptide containing Glu in the N-terminus was incubated under the same conditions it too was not a substrate for arginylation. These findings show that neither the presence of an N-terminal acidic residue nor an oxidized residue alone are sufficient to signal arginylation. Thus, another feature of the oxidized A-chain of insulin is required for arginylation. That feature remains to be identified. PMID- 10685605 TI - Differential association of tau with subsets of microtubules containing posttranslationally-modified tubulin variants in neuroblastoma cells. AB - Neuronal cells display different subsets of dynamic microtubules. In axons and extending neurites, this intrinsic dynamics is modulated by the microtubule associated protein tau. Moreover, posttranslational modifications of tubulin, namely acetylation, tyrosination or glutamylation are directly involved in determining the stability of neuronal microtubules. Studies were carried out to analyze the interaction patterns of tau with subsets of microtubules in N2A neuroblastoma cells, which can differentiate in the presence of dibutyryl cAMP. Double labeling studies showed a differential pattern of tau association with microtubules containing acetylated and tyrosinated tubulin. Furthermore, studies using depolymerizing drugs revealed a selectivity in the association of tau with microtubular polymers and microfilaments, within the organization of the neuronal cytoskeleton. In order to study the association of specific tau isoforms with microtubules containing modified tubulin variants, immunoprecipitation studies were carried out. The coimmunoprecipitation data indicated a selective binding of specific tau isoforms to either modified tubulin variant. To assess the hypothesis on the roles of tau isoforms in the stabilization of microtubules containing modified tubulins, the association of those variants with tau isoforms was analyzed in overlay experiments. A preferential binding of acetylated tubulin from undifferentiated N2A cell extracts, to at least one slow-migrating tau isoform was revealed. However, acetylated tubulin from N2A cells containing long neurites displayed a preferential association with two isoforms of tau. On the other hand, tyrosinated tubulin from N2A extracts bound to the entire set of neuronal tau isoforms. These studies, along with the tau association with microtubules with different stability, indicate that tau segregates into subsets of microtubules in the axonal processes. The studies also suggest that these interactions may respond to a functional versatility of these polymers in differentiating neurons. PMID- 10685606 TI - Post-translational arginylation of proteins in cultured cells. AB - The aim of this study was to analyze the N-terminal post-translational incorporation of arginine into cytosolic proteins from cultured cells and the in vitro incorporation of arginine into soluble proteins of PC12 cells after serum deprivation. Arginine incorporation was measured in the presence of protein synthesis inhibitors. None of the inhibitors used affected significantly the arginylation reaction while the novo synthesis of protein was reduced by 98%. Under these conditions, we found that of the total [14C]arginine incorporated into the proteins, around 20% to 40% was incorporated into the N-terminal position of soluble proteins by a post-translational mechanism. These results suggest that this post-translational aminoacylation may be a widespread reaction in neuronal and non-neuronal cells. We also found that in PC12 cells, the in vitro post-translational arginylation was 60% higher in apoptotic cells with respect to control cells. These findings suggest that the post-translational arginylation of proteins may be involved in programmed cell death. PMID- 10685607 TI - Epifluorescence microscopy of surface domain microheterogeneity in myelin monolayers at the air-water interface. AB - Myelin lipids form liquid-expanded monolayers at the air-water interface, with no evidence of surface pressure-induced two-dimensional phase transition. However, the film doped with 2 mole % of the fluorescent probe N-(7-nitro-2-1,3 benzoxadiazol-4-yl) Diacyl Phosphatidyl-ethanolamine (NBD-PE) shows an irregular pattern of coexisting laterally segregated surface domains with diffuse boundaries that change from smooth patterns to fractal-like structures depending on surface pressure. Successive expansion-recompression cycles lead to more defined domains, with a general reorganization occurring at surface pressures of about 20 mN/m. At least two coexisting phases occur over almost all the compression isotherms. The presence of proteins in whole myelin monolayers induces defined domain textures with relatively sharp boundaries. The patterns during compression and expansion are quite similar and, after the first cycle, little changes occur under recompression. The patterns observed provide topographical evidence for the existence of dynamic domain microheterogeneity in the surface of myelin interfaces. PMID- 10685608 TI - Myelin membranes isolated from rats intracranially injected with apotransferrin are more susceptible to in vitro peroxidation. AB - Purified myelin obtained from 17 day old rats intracranially injected with aTf at 3 days of age was submitted to in vitro peroxidation using Fe + ascorbic acid (FeA) or Cu + H2O2 (CuH), to investigate the susceptibility of this membrane to in vitro metal catalyzed peroxidation. There was an increase in thiobarbituric acid-reactive-substances (TBARS) (60%) and in protein-associated carbonyls (PAC) (20%) in the myelin from aTf injected rats in comparison to myelin from controls, indicating a higher susceptibility to peroxidation. Desferoxamine (DFX) injected simultaneously with aTf did not change the response of myelin to aTf. CNS myelin is highly vulnerable to oxidative stress, and its susceptibility to peroxidation increases in myelin isolated from aTf injected rats. This increased liability to peroxidation as well as the previously reported aTf-dependent increment in certain myelin proteins and lipids and in the expression of specific myelin mRNAS, does not appear to be due to an increased amount of iron bound to the injected aTf. The changes in composition that we have previously reported could result in an abnormal myelin, allowing the peroxidative system to act upon the membrane more easily than in normal circumstances. PMID- 10685609 TI - Regulation of signaling by protein-tyrosine phosphatases: potential roles in the nervous system. AB - During neuronal development, cells respond to a variety of environmental cues through cell surface receptors that are coupled to a signaling transduction machinery based on protein tyrosine phosphorylation and dephosphorylation. Receptor and non-receptor tyrosine kinases have received a great deal of attention; however, in the last few years, receptor (plasma membrane associated) and non-receptor protein-tyrosine phosphatases (PTPs) have also been shown to play important roles in development of the nervous system. In many cases PTPs have provocative distribution patterns or have been shown to be associated with specific cell adhesion and growth factor receptors. Additionally, altering PTP expression levels or activity impairs neuronal behavior. In this review we outline what is currently known about the role of PTPs in development, differentiation and neuronal physiology. PMID- 10685610 TI - Multiple forms of phosphatase from human brain: isolation and partial characterization of affi-gel blue binding phosphatases. AB - Implication of protein phosphatases in Alzheimer disease led us to a systemic investigation of the identification of these enzyme activities in human brain. Human brain phosphatases eluted from DEAE-Sephacel with 0.22 M NaCl were resolved into two main groups by affi-gel blue chromatography, namely affi-gel blue binding phosphatases and affi-gel blue-nonbinding phosphatases. Affi-gel blue binding phosphatases were further separated into four different phosphatases, designated P1, P2, P3, and P4 by calmodulin-Sepharose 4B and poly-(L-lysine) agarose chromatographies. These four phosphatases exhibited activities towards nonprotein phosphoester and two of them, P1 and P4, could dephosphorylate phosphoproteins. The activities of the four phosphatases differed in pH optimum, divalent metal ion requirements, sensitivities to various inhibitors and substrate affinities. The apparent molecular masses as estimated by gel filtration for P1, P2, P3, and P4 were 97, 45, 42, and 125 kDa, respectively. P1 is markedly similar to PP2B from bovine brain and rabbit skeletal muscle. P4 was labeled with anti-PP2A antibody and may represent a new subtype of PP2A. P1 and P4 were also effective in dephosphorylating Alzheimer disease abnormally hyperphosphorylated tau (AD P-tau). The resulting dephosphorylated AD P-tau had its activity restored in promoting assembly of microtubules in vitro. These results suggest that P1 and P4 might be involved in the regulation of phosphorylation of tau in human brain, especially in neurodegenerative conditions like Alzheimer's disease which are characterized by the abnormal hyperphosphorylation of this protein. PMID- 10685611 TI - Kinetics of Na+, K+-ATPase inhibition by an endogenous modulator (II-A). AB - We have previously reported the isolation by gel filtration and anionic exchange HPLC of two brain Na+, K+-ATPase inhibitors, II-A and II-E, and kinetics of enzyme interaction with the latter. In the present study we evaluated the kinetics of synaptosomal membrane Na+, K+-ATPase with II-A and found that inhibitory activity was independent of ATP (2-8 mM), Na+ (3.1-100 mM), or K+ (2.5 40 mM) concentration. Hanes-Woolf plots showed that II-A decreases Vmax in all cases; KM value decreased for ATP but remained unaltered for Na+ and K+, indicating respectively uncompetitive and noncompetitive interaction. However, II A became a stimulator at 0.3 mM K+ concentration. It is postulated that brain endogenous factor II-A may behave as a sodium pump modulator at the synaptic region, an action which depends on K+ concentration. PMID- 10685612 TI - Two glycogen synthase activities associated with proteoglycogen in retina. AB - Glycogen synthase of bovine retina was found associated with the acid-insoluble and acid-soluble proteoglycogen fractions. The synthase associated with the acid insoluble proteoglycogen precursor showed an 8-fold lower Km for UDP-glucose than the synthase associated with the acid-soluble fraction, and was inhibited by detergent. A short digestion with pronase resulted in conversion of the acid insoluble fraction into acid-soluble. The results lead us to postulate that the acid-insolubility of the proteoglycogen fraction and the association with retina membrane proposed before, is caused by glycogen synthase strongly associated to its polysaccharide moiety. The enlargement of the polysaccharide moiety during proteoglycogen biosynthesis, from glycogenin linked to a few 11 to 12 glucose units to the acid-insoluble proteoglycogen precursor (Mr 470,000) would be carried out, together with the branching enzyme, by the glycogen synthase showing a low Km for UDP-glucose. The glycogen synthase with the highest Km for UDP glucose would participate in conversion of the precursor into mature acid-soluble proteoglycogen. PMID- 10685613 TI - Scinderin, a Ca2+-dependent actin filament severing protein that controls cortical actin network dynamics during secretion. AB - Secretory vesicles are localized in specific compartments within neurosecretory cells. These are different pools in which vesicles are in various states of releasability. The transit of vesicles between compartments is controlled and regulated by Ca2+, scinderin and the cortical F-actin network. Cortical F-actin disassembly is produced by the filament severing activity of scinderin. This Ca2+ dependent activity of scinderin together with its Ca2+-independent actin nucleating activity, control cortical F-actin dynamics during the secretory cycle. A good understanding of the interaction of actin with scinderin and of the role of this protein in secretion has been provided by the analysis of the molecular structure of scinderin together with the use of recombinant proteins corresponding to its different domains. PMID- 10685614 TI - Glucosylceramide synthesized in vitro from endogenous ceramide is uncoupled from synthesis of lactosylceramide in Golgi membranes from chicken embryo neural retina cells. AB - It is known that ceramide (Cer), the precursor of sphingoglycolipids and of sphingomyelin, participates in events leading to activation of the apoptotic pathway, and per se or through conversion to glucosylceramide (GlcCer) modulates formation of neuritic processes in developing neurons. To learn about the fate of de novo synthesized Cer and GlcCer we examined, in Golgi membranes from chicken embryo neural retina cells, the metabolic relationships of endogenous Cer, GlcCer and lactosylceramide (LacCer). Incubation of the membranes with UDP-[3H]Glc revealed a pool of endogenous Cer useful for synthesis of GlcCer. Most of the GlcCer synthesized, however, was not used for synthesis of LacCer, indicating that it was functionally uncoupled from LacCer synthase. On the other hand, incubation with UDP-[3H]Gal revealed a pool of endogenous GlcCer that depending of the integrity of the membranes was functionally coupled to LacCer and ganglioside synthesis. These results indicate that most GlcCer formed in vitro from Cer is topologically segregated from the synthesis of LacCer. However, subfractionation in sucrose gradients of Golgi membranes labeled with both precursors failed to separate membranes enriched in [3H]GlcCer from those enriched in [3H]Gal-labeled LacCer. It is concluded that despite both transfer steps co-localize in the Golgi membranes, coupling of GlcCer synthesis to LacCer synthesis requires conditions not present in our in vitro assay. This suggests that a coupling activity exists that could be relevant for regulation of the cytoplasmic levels of Cer and GlcCer. PMID- 10685616 TI - Ganglioside expression during differentiation of chick retinal cells in vitro. AB - The neural retina has been widely used to study the developmental patterns of ganglioside metabolism. Recent findings about in vitro differentiating chick embryo retina cells showed that: a) GD3 and GD1a ganglioside patterns undergo the most dramatic changes; b) when the cells emit neurites, GD3 ganglioside and a group of complex gangliotetraosylgangliosides (GTOG) are transiently coexpressed; c) synchronized developmental phenomena are dissociated by anti-GM1 antibodies; d) GD3 remains as a major ganglioside in differentiated neurons, though it is almost not immunoexpressed; e) GTOG affect antibody binding to GD3; f) the content of gangliosides involved in neural differentiation modifies their immunostain localization on cell membrane; g) after exogenous GTOG uptake, immature neurons mimic GD3 immunofluorescent localization of mature cells; h) a subset of purified retinal ganglion cells express GTOG characteristic of mature neurons. PMID- 10685615 TI - Immediate early gene expression within the visual system: light and circadian regulation in the retina and the suprachiasmatic nucleus. AB - Immediate early genes are a family of genes that share the characteristic of having their expression rapidly and transiently induced upon stimulation of neuronal and non-neuronal cells. In this review, first a short description of the IEGs is given, then it is discussed the stimulus-induced and circadian-induced variations in the expression of IEGs in the visual system, mainly in the retina and the suprachiasmatic nucleus. The possible physiological consequences of these variations in IEG expression are also considered. Finally, we refer to two aspects of our recent studies and those of other laboratories involving light driven IEG expression. The first is the finding that in the chick retina, the expression of c-fos is differentially modulated in the different cell types and that c-fos regulates the synthesis of the quantitatively most important lipids of all cells, the phospholipids, by a non-genomic mechanism. The second is the occurrence of differential waves of IEG expression in the mammalian suprachiasmatic nucleus regarding light induction or spontaneous oscillations. PMID- 10685618 TI - The effectiveness of rehabilitation: a critical review of the evidence. Introduction. PMID- 10685617 TI - Expression of a neuronal nicotinic acetylcholine receptor in insect and mammalian host cell systems. AB - Different mammalian and insect somatic host cell systems were tested in their ability to express, fold, and assemble alpha7-type neuronal acetylcholine receptor (AChR) both at the transcriptional and translational level. For this purpose we employed clonal cell lines derived from the neural crest, such as PC12 cells from a rat adrenal pheochromocytoma, and GH3 cells isolated from a rat pituitary tumor, as well as non-neuronal cells such as NIH-3T3 fibroblasts from embryonic NIH Swiss mouse and Sf9 cells from ovary tissue of the Spodoptera frugiperda butterfly. Total RNA, isolated from either transfected or non transfected PC12, GH3 or 3T3 cells, or recombinant AcNPV-infected and mock infected Sf9 cells was analyzed by Northern blot. PC12 cells, which endogenously express alpha7 AChR, and all its heterologous alpha7-transfectant clones, exhibited variable but generally high amounts of a single transcript. GH3 and NIH 3T3 transfectant clones and recombinant AcNPV-infected Sf9 cells expressed variable levels of alpha7-mRNA, with a single transcript that co-migrated with the 28S rat rRNA. Only the neural crest-derived cell lines appeared to functionally express the alpha7 AChR, as measured by their [125I]alpha bungarotoxin binding ability. The results suggest that heterologous expression of alpha7 is regulated not at the transcriptional, but at the postranslational level and that not all host cell systems appear to express the cellular factors needed for the correct postranslational modifications leading to mature and functional alpha7 AChR. Furthermore, the results suggest that tightly controlled expression mechanisms have evolved in parallel with this ancient cholinergic sequence. PMID- 10685619 TI - Effectiveness of brain injury rehabilitation. AB - Despite the problems posed by diversity of condition and the lack of agreement among researchers over what outcome to measure, there is now increasingly robust evidence for the effectiveness of rehabilitation in brain-injured populations. Meta-analysis has demonstrated clearly that stroke units provide a better outcome than management on a general medical ward, at the level of survival, discharge destination and dependency. The extent of this advantage may be summarized in the following terms. For every 100 patients treated in a stroke unit, four deaths and two institutional admissions are avoided, and five patients are discharged home. This benefit appears to arise from a combination of good-quality acute management and the coordinated input of a multidisciplinary team. Therapy programmes are shown to be of benefit and intensive therapy programmes of somewhat greater benefit. Smaller numbers and heterogeneity among the head-injured population tend to confound randomized controlled trial designs, but there is no good reason to suppose that brain injury resulting from trauma should be less responsive to similar good management principles than that arising from stroke. In any event, we have progressed to a stage where the weight of evidence supports the notion that rehabilitation is effective, and nontreatment controls are ethically no longer acceptable. It is time now to unravel the threads of rehabilitation and consider which are the critical components. There are still many opportunities for comparison of different models for delivery of care, and the existing evidence for these is discussed. PMID- 10685620 TI - Effectiveness of spinal cord injury rehabilitation. AB - As in other areas of rehabilitation, relatively small numbers and diversity--both of condition and of patients' goals--hinder the assimilation of robust evidence for the effectiveness of rehabilitation. Patients with spinal cord injury (SCI) tend to be gathered together in a small number of regional services, each with their own philosophy and each with different attitudes to outcome measurement, and thus collection of the existing trials for meta-analysis is problematic. The marked improvement in outcome from SCI that has occurred with the development of specialist rehabilitation programmes argues strongly for the effectiveness of rehabilitation, and we have progressed beyond the point where randomized controlled trials that deny a group such intervention could be considered ethical. Current research is aimed at teasing apart the aspects of different care models that are most effective, or the evidence for the usefulness of interventions for control of symptoms such as spasticity and pain. This evidence is reviewed and discussed. PMID- 10685621 TI - Effectiveness of rehabilitation for multiple sclerosis. AB - Unlike other areas of rehabilitation, which typically follow a single incident such as trauma or stroke and are followed by improvement (or at least an expectation of stable impairment), multiple sclerosis (MS) presents the problem of progressive impairment and disability. In addition the nature and course of this progression are variable, so that the population is heterogeneous. Expectations for outcome must be modest, and measurement should be focused on quality on life issues. On a background of pre-existing complex disability, multiple single-case (before and after) study designs often present the best evidence for effectiveness of the team approach and for specific interventions. This evidence is presented and reviewed. PMID- 10685622 TI - Effectiveness of rehabilitation following amputation. AB - For many patients, the outcome of rehabilitation following amputation depends substantially on comorbidity. This is particularly so where amputation is performed because of peripheral vascular disease which may involve other end organs, but also applies in trauma where loss of limb may not be the only injury. In evaluating outcome, measures must take account of the very different goals for rehabilitation. These may range from cosmesis or the simple ability to transfer from bed to chair, to successful competition in the Para-Olympics. Rehabilitation programmes for amputees are not simply prosthetic services, but must take account of the whole patient, their goals and ambitions. Research to date has made a contribution in identifying prognostic factors for prosthetic rehabilitation, thus helping to target limited resources. Controlled studies are still required, however, to establish the optimum services to offer to different groups of patients. PMID- 10685623 TI - Effectiveness of rehabilitation in arthritis. AB - If rehabilitation aims to improve function, the demonstration of its effectiveness requires functional assessment, rather than the standard clinical and laboratory tests beloved of many rheumatologists. Medical interventions are not often evaluated for their contribution to improved function, and this omission must be addressed in the future. The evidence, where it is available, supports a multidisciplinary approach, but increasingly stresses the importance of partnership with the patient, passing back to them the responsibility for maintaining their own exercise programmes and involvement in activities. However, the provision of background support appears to be essential to continued independence. Evidence for the various interventions is reviewed. PMID- 10685625 TI - The effectiveness of rehabilitation: a critical review of the evidence. Closing remarks PMID- 10685624 TI - Effectiveness of rehabilitation for spinal pain. AB - The evidence for effectiveness of different approaches is often diluted by the inclusion of heterogeneous groups and, in this case, lack of agreement over what constitutes 'acute' and 'chronic' back pain is a clear confounding factor. Although there are undoubtedly common issues in the approach to these problems, there are also clear differences. Closer definition of the problem and the development of specific and targeted outcome measures is required. In common with other areas of musculoskeletal rehabilitation, the evidence strongly supports exercise (except possibly in the rare case of true radicular back pain) and a cognitive behavioural approach to pain management. The variability of evidence in support of manipulation suggests that patient selection is important, but as yet those selection criteria are not clear. As is often the case, medical interventions are rarely submitted to evaluation in terms of functional outcome so, for procedures such as epidurals and facet joint injections, the jury is still out at the current time. PMID- 10685626 TI - The Consensus Conference on the treatment of in situ ductal carcinoma of the breast, April 22-25, 1999. PMID- 10685627 TI - Intramucosal cysts in the gastric body of patients with Zollinger-Ellison syndrome. AB - To ascertain the frequency and the clinico-functional correlations of intramucosal cysts in the gastric body of patients with the Zollinger-Ellison syndrome (ZES) and to clarify the relevant mechanism of development, a total of 106 consecutive ZES patients (58 M, 48 F; mean age: 53 yrs, range 19-93 yrs) were investigated with a mean of 7.2 biopsy specimens of the body mucosa per patient proved to be suitable for the study. Biopsies of endoscopically detectable polypoid lesions were not considered. Cystic changes were evaluated with respect to their severity by assessing the cyst grade (0, absent, 1; <30%, 2; 30-60%; 3 >60% of the mucosal area of the biopsy specimen of individual patients showing the most pronounced finding, respectively) and to their intragastric distribution by assessing the ratio of biopsy specimens showing cystic changes over the total number of biopsies examined in each patient. Intramucosal cysts were found in biopsies of non-polypoid gastric body mucosa in 71.7% of 106 patients with Zollinger-Ellison syndrome (ZES) and showed grade 2 and 3 severity in 22 and 8 cases, respectively. The severity of cystic changes correlated with the gastrin levels (p = 0.0005) and was more advanced in patients with active than in those with cured disease (p = 0.037). In the former group, furthermore, advanced cystic changes correlated with age (p = 0.03), male gender (p = 0.014), years of disease from onset (p < 0.02), years of omeprazole treatment (p = 0.033), basal acid output (p < 0.02), severity of ECL cell proliferative changes (p = 0.028), and absence of previous gastrinoma resection (p = 0.039) whereas they did not correlate with MEN-1 status, gastritis, maximal acid output, total duration of any antisecretory drug treatment, daily doses of omeprazole (> 20 mg vs 20 mg), years from surgery, duodenal localization of gastrinoma(s), presence of gastric carcinoid tumor(s) and of liver metastases. In groups of patients subdivided according to three levels of serum gastrin, the duration of omeprazole treatment was not related to the severity of cystic changes. It is concluded that intramucosal cysts in non polypoid gastric body mucosa of ZES patients are by far more common than the already reported fundic gland polyps, to which they likely give raise. Circulating levels of gastrin have an important independent role in their development. PMID- 10685628 TI - Cells producing cathepsins D, B, and L in human breast carcinoma and their association with prognosis. AB - Lysosomal proteinases, cathepsins D, B, and L have been associated with malignant tumor progression and with prognosis in various human carcinomas. In the current study, the immunohistochemical localization of cathepsins in tumor cells was correlated with cathepsin protein concentration in breast carcinoma cytosols from 77 patients. Significant correlation was found for cathepsin D (P < .041) and borderline correlation for cathepsin B (P < .055) but not for cathepsin L. We hypothesize that the poor correlation of cysteine cathepsins was attributable to the fact that they were present not only in malignant epithelial cells, but also in infiltrating macrophages and stromal fibroblasts. In addition, tumor surrounding myoepithelial cells (42% of tumors) and myofibroblasts (26% of tumors) as well as endothelial cells of neovasculature (10% of tumors) all stained specifically for cathepsin B. Two thirds of tumors co-expressed cathepsins B and L in tumor cells, whereas only 17% of tumors co-expressed all 3 cathepsins. Intense immunostaining for cathepsin D of tumor cells was observed in tumors at high TNM stage and tumors having positive lymph nodes. The expression of cathepsin B was independent of established prognostic factors, whereas intense cathepsin L staining in tumor cells was associated with high histological grade. With respect to prognosis of patient survival, only tumor cell-associated cathepsin D (P = .042) and myoepithelial cell-associated cathepsin B (P = .061) showed borderline significance. Cathepsins B and L immunostaining in tumor cells was not prognostic. In contrast, cytosolic levels of cathepsin B correlated with higher rate of relapse. Taken together, these results show the diversity in the cellular distribution of cathepsins in human breast carcinoma, presumably reflecting specific regulation and function of each of the cathepsins during tumor progression. PMID- 10685629 TI - Unusual bone marrow manifestations of parvovirus B19 infection in immunocompromised patients. AB - Parvovirus B19 is responsible for a spectrum of disease in humans. The usual bone marrow findings in acute parvovirus infections are marked erythroid hypoplasia and occasional giant erythroblasts. Intranuclear inclusions in developing erythroid precursors are rarely described in children or adults with parvovirus infection, although abundant intranuclear inclusions are commonly observed in the placenta and other tissues in infected fetuses. In this study, 8 patients are reported in whom the first evidence of parvovirus infection was the recognition of numerous intranuclear inclusions in erythroid precursors on bone marrow biopsy sections. Six of the 8 patients had documented immunodeficiencies; 4 had acquired immune deficiency syndrome (AIDS), and 2 were on chemotherapy. Five of 7 patients were negative for immunoglobulin G (IgG) antiparvovirus antibodies, including all 4 with AIDS. Unlike the typical pattern in parvovirus infection, the bone marrow was hypercellular in most of the patients, and erythroid precursors were usually increased with the entire spectrum of normoblast maturation represented; abundant intranuclear inclusions were observed similar to the finding in fetuses. The inclusions were variably eosinophilic and compressed the chromatin against the nuclear membrane. In situ hybridization showed parvovirus B19 DNA in numerous erythroid precursors in all cases. The findings of erythroid maturation and abundant viral inclusions in these immunocompromised patients is consistent with the hypothesis that failure to produce effective IgG parvovirus neutralizing antibodies may lead to persistent infection through viral tolerance that allows erythroid development of infected cells past the pronormoblast stage. Identification of parvovirus inclusions in marrow biopsies and subsequent confirmation of infection by in situ hybridization can be important in the assessment of anemia in immunodeficient patients because serological studies for parvovirus B19 are frequently negative. PMID- 10685630 TI - Frequent c-myc and Int-2 overrepresentations in nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is a commonly occurring tumor in southern China. Although several causative factors have now been recognized, the genetic basis underlying its tumorigenesis is still unclear. To identify potential chromosomal aberrations for further investigations, comparative genomic hybridization (CGH) analysis was applied to the study of genomic imbalances in 10 NPC biopsy specimens. Before CGH analysis, the tumor cell content within the biopsy specimens was enriched by tissue microdissection, and universal genome amplification was performed on the recovered DNA. Recurrent chromosomal gains were detected on 1q (6 of 10 cases), 2q (5 of 10 cases), 3q (7 of 10 cases), 6p (8 of 10 cases), 6q (5 of 10 cases), 7q11.2 (4 of 10 cases), 8q (6 of 10 cases), 11q13, 12, and 15q (8 of 10 cases each), 17q (6 of 10 cases), and 20q (5 of 10 cases). Common losses were identified on 3p (5 of 10 cases), 9p (5 of 10 cases), 11q14-qter (8 of 10 cases), and 14q (5 of 10 cases). Among these aberrations, 7, 8, and 11 gains were further investigated on a series of NPC tissue samples, by interphase fluorescent in situ hybridization (FISH), for the incidence of alpha satellites: 7, 8, and 11 c-myc and Int-2. Low-level increases of alpha-satellite 7 (9 of 34 cases; 26.5%), alpha-satellite 8 (15 of 34 cases; 44%), and alpha satellite 11 (8 of 32 cases; 25%) were detected, whereas high-level copy gains of c-myc (21 of 34 cases; 62%) and Int-2 (26 of 34 cases; 76.5%) were more frequently found. Our series is the first to identify genomic overrepresentations of c-myc and Int-2 in NPC. The high incidence of Int-2 amplifications strongly suggests a role of this proto-oncogene in the pathogenesis of NPC. PMID- 10685631 TI - Expression of Bcl-2 familial proteins is reduced in small bile duct lesions of primary biliary cirrhosis. AB - In primary biliary cirrhosis, biliary epithelial cell death by apoptosis results in progressive bile duct loss. We examined immunohistochemically 4 apoptosis regulating bcl-2 familial proteins (bcl-2, mcl-1, bcl-X, and bax) in the biliary epithelium in 19 cases of primary biliary cirrhosis. Ten cases of chronic hepatitis C, 9 cases of extrahepatic biliary obstruction, and 10 cases of normal liver were used as a control. Bcl-2 and mcl-1 are inhibitors of apoptosis, bcl-X, probably bcl-XL in biliary epithelial cells, an inhibitor, and bax, a promoter of apoptosis. First, we clarified the distribution of bcl-2 familial proteins on the intrahepatic biliary tree in normal livers. Bcl-2 was detected in the interlobular bile ducts and bile ductules, but not in the large and septal bile ducts in all cases examined. Mcl-1, bcl-X, and bax were diffusely detectable at the any level of the intrahepatic biliary tree, with a staining pattern that was diffuse and cytoplasmic. This distribution pattern was preserved in extrahepatic biliary obstruction. Bcl-2 expression was lost or markedly reduced in the damaged interlobular bile ducts in primary biliary cirrhosis, whereas the reduction was only focal or mild in the bile ducts with hepatitis-associated damage in chronic hepatitis C. Expression levels of mcl-1, bcl-X, and bax were similarly reduced to that of bcl-2 in these 2 diseases. These findings suggest that bax is not important as a proapoptotic factor in the damaged bile ducts and that downregulation of bcl-2 and mcl-1, and probably that of bcl-XL, leads to a decrease in the threshold of apoptosis and increase in the vulnerability to apoptotic stimuli in these bile ducts, followed by the progressive apoptotic loss of interlobular bile ducts, in primary biliary cirrhosis. PMID- 10685632 TI - Expression of hepatocyte growth factor (HGF)/scatter factor and its receptor c MET correlates with poor prognosis in synovial sarcoma. AB - The hepatocyte growth factor (HGF)/c-MET signaling system plays an important role in the carcinogenesis of various organs. We investigated the expression of HGF and its receptor c-MET by immunohistochemistry (IHC) in 69 cases of synovial sarcoma and compared the findings with clinicopathologic parameters, proliferating activities evaluated by MIB-1 labeling index (MIB-1 LI), and patients' prognosis. Furthermore, mRNA analysis of HGF, c-MET, and SYT-SSX fusion gene was performed by reverse transcriptase-polymerase chain reaction (RT-PCR) in 22 concordant frozen materials. Twenty-one of 69 (30.4%) tumors showed positive reaction for c-MET, whereas 22 tumors (31.9%) were positive for HGF. In 10 cases, co-expression of HGF and c-MET was observed; however, there was no significant correlation between HGF and c-MET expression. HGF expression was correlated with female patients, large tumors (more than 5 cm), the presence of rhabdoid cells, low frequency of mast cells (<20/10 HPF), high nuclear grade (grade III), and high American Joint Committee (AJC) stage (III and IV). Conversely, c-MET expression was only correlated with large tumors. However, the coexpression of HGF and c-MET was significantly correlated with large tumor size, the existence of rhabdoid cells, and high AJC stage. Both the expression of HGF and the co expression of HGF and c-MET showed a significantly high MIB-1 LI and were correlated with poor prognosis according to univariate analysis. Multivariate Cox analysis showed that high AJC stage, the expression of HGF, and a high MIB-1 LI (12.0>) independently had a negative impact on overall survival. In 22 frozen material cases evaluated by both IHC and RT-PCR, a statistically significant correlation was found between the 2 techniques. SYT-SSX fusion transcripts were detected in all 22 cases. Three tumors had SYT-SSX2 fusion transcripts, whereas 19 had SYT-SSX1 phenotype. Our results suggest that HGF/c-MET paracrine signaling may contribute to tumorigenesis and progression in synovial sarcoma. PMID- 10685633 TI - Expression of matrix metalloproteinase-2 (MMP-2) and of membrane-type-1-matrix metalloproteinase (MT1-MMP) in transitional cell carcinoma of the upper urinary tract. AB - Tumor cell invasion and metastasis are biologically dependent on the proteolytic destruction of surrounding matrix components. Matrix metalloproteinase-2 (MMP-2) is able to cleave type IV collagen, and membrane-type-1-matrix metalloproteinase (MT1-MMP) induces activation of proMMP-2. We investigated the expression of MMP-2 and MT1-MMP using in situ hybridization and immunohistochemistry in 102 cases of transitional cell carcinoma of the upper urinary tract (TCC-UUT). A positive expression of each metalloproteinase was recognized in all samples and was apparent within the cytoplasm of tumor epithelial cells and/or stromal cells situated at the interface between tumor and stroma. Our analysis of clinicopathologic findings showed a relationship between MMP-2 and MT1-MMP expression and stage. The correlation between the MMP-2 protein staining score for tumor epithelial cells and overall survival rate reached significance in the univariate analysis. However, only stage was associated with disease-free and overall survivals in the multivariate analysis. In conclusion, the detection of MMP-2 and MT1-MMP would appear to be of limited value in informing the prognosis of TCC-UUT. PMID- 10685634 TI - Histopathologic analysis of chromosome aneuploidy in ductal carcinoma in situ. AB - Formalin-fixed, paraffin-embedded sections from 28 cases of ductal carcinoma in situ (DCIS; 12 with coexisting invasive neoplasm) were analyzed for numerical alterations of chromosomes 7, 8, 16, and 17 by performing fluorescence in situ hybridization (FISH) using centromeric (alpha-satellite) probes. Based on signal counts in 200 to 300 nuclei, each hybridization was classified as disomic (copy loss in <40%, copy gain in < 10%), monosomic (copy loss in at least 50% of nuclei, partial if 40% to 49%) or trisomic/polysomic (copy gain in at least 20% of nuclei, partial if 10% to 19%). Grade I lesions were characterized by complete lack of significant chromosome gain, but 29% showed partial (focal) monosomy. Grade III lesions, in contrast, showed partial or complete trisomy/polysomy in 88% of hybridizations versus monosomy in only 4%. Grade II DCIS exhibited a mixed pattern of chromosome aneuploidy: 38% hybridizations were disomic, 36% trisomic/polysomic, and 26% monosomic (8 of 10 hybridizations showing complete monosomy occurred in grade II lesions). Disomic hybridizations exhibiting rare cells (5% to 10%) with copy gain were more frequent in tumors with coexisting invasive neoplasm (5 of 17 v 2 of 33, P = .02). In morphologically heterogeneous lesions, higher-grade foci were characterized by chromosome copy gain relative to corresponding lower-grade areas in 17 of 22 (77%) hybridizations. These results show the presence of multiple (at least 3) distinct chromosome aneuploidy patterns in DCIS, in keeping with divergent mechanisms of genetic alteration. Degree of chromosomal instability, moreover, may correlate with progression of DCIS to invasive growth, implying that genetic instability is a parameter that impacts the likelihood of early breast carcinoma progression. PMID- 10685635 TI - Sarcomatoid salivary duct carcinoma of the parotid gland. AB - Salivary duct carcinoma (SDC) is a high-grade neoplasm known to histologically resemble high-grade ductal carcinoma in situ of the breast. We describe 3 cases of sarcomatoid salivary duct carcinoma, a heretofore unreported variant of SDC. Each case was a composite of SDC and sarcomatoid carcinoma and histologically similar to reported cases arising in the breast. The clinicopathologic features, including immunohistochemistry, of 3 cases were investigated. In the 3 men, ages 56, 68, and 70 years, the resected parotid tumors measured 1.5, 3.5, and 1.5 cm, respectively. Only the 3.5-cm tumor extended beyond the parotid gland into soft tissue. This patient died at 3 years with pulmonary metastases. The other patients were free of disease at 6 and 12 months. Histologically, each case was a composite of usual-type SDC and sarcomatoid carcinoma. SDC showed typical cribriform architecture, whereas anaplastic, spindled cells constituted the sarcomatoid areas. Immunohistochemically, epithelial elements stained as follows: cytokeratin (AE1/AE3 & CAM 5.2) positive in 3 of 3 cases, EMA positive in 3 of 3 cases, vimentin negative in 3 of 3 cases, desmin negative in 3 of 3 cases, c-erbB 2 positive in 1 of 2 cases. Sarcomatoid elements stained as follows: AE1/AE3 negative in 3 of 3 cases, CAM 5.2 rare positive cell in 1 of 3 cases, EMA focally positive in 3 of 3 cases, vimentin positive in 3 of 3 cases, desmin negative in 3 of 3 cases, c-erbB-2 negative in 2 of 2 cases. Electron microscopy, performed in one case, showed scattered junctional complexes congruent with epithelial differentiation. Immunohistochemical results, EMA and CAM 5.2 positivity, and ultrastructural findings supported our belief that these unique biphasic tumors represented SDC with sarcomatoid carcinoma. We conclude an element of sarcomatoid carcinoma rarely may arise in association with SDC, and it is erroneous to diagnose such tumors as "carcinosarcoma." PMID- 10685636 TI - Expression of DNA topoisomerase I in neoplasms of the kidney: correlation with histological grade, proliferation, and patient survival. AB - Renal cell carcinoma is an inherently chemotherapeutically resistant neoplasm. Because of this, new drugs targeting this tumor are needed. One class of new anticancer drug targets the enzyme DNA topoisomerase I. Laboratory data indicate that cells sensitive to topo I targeting drugs contain high topo I levels. To determine whether some renal cell carcinomas contain elevated topo I and might therefore be targets of topo I active antitumor agents, we used a new immunohistochemical stain for topo I to determine the expression of the enzyme in 51 tumors of the kidney. Increased topo I expression was found in 4 of 11 (36%) grade 3 renal cell carcinomas and in 8 of 8 (100%) grade 4 renal cell carcinomas. Normal topo I expression was observed in all adenomas, oncocytomas, and grade 1 and grade 2 renal cell carcinomas. Because topo I targeted anticancer drugs are S phase specific, topo II-alpha and MIB-1 proliferation indices also were performed. Topo II-alpha correlates well with MIB-1 (correlation coefficient = 0.96). Of the 12 tumors with elevated topo I, only 3 had topo II-alpha proliferation indices greater than 40, indicating a tumor with elevated topo I expression and a large growth fraction. We hypothesize that these tumors might be susceptible to topo I anticancer drug therapy. In addition, we found that the average topo II-alpha proliferation index of tumors from patients who died of disease was 27.4 +/- 19.8, which was statistically different from the average topo II-alpha index of 5.8 +/- 6.5 observed in tumors from patients who remained alive during our follow-up. PMID- 10685637 TI - Lack of CD 29 (beta1 integrin) and CD 54 (ICAM-1) adhesion molecules in intravascular lymphomatosis. AB - Intravascular Lymphomatosis (IL) is a rare and usually aggressive form of non Hodgkin's lymphoma characterized by the growth of neoplastic cells within vascular lumina that usually presents with skin or central nervous system (CNS) involvement. The mechanism(s) for the selective intravascular growth of this neoplasm remain(s) unexplained. We now report clinical and immunohistologic data on surgical material from 6 cases of IL; in 4 of 6 cases, autopsies were performed. Our IL cases shared the following features: (1) B-cell lineage; (2) lack of skin involvement at presentation; (3) aggressive behavior; and (4) lack of extravascular lymphomatous masses; in addition, 1 case had an associated gastric low-grade MALT lymphoma. We studied by immunohistochemistry formalin fixed, paraffin-embedded sections with monoclonal antibodies to molecules known to be involved in lymphocyte and endothelial adhesion phenomena, that is, CD29 (beta1 integrin subunit), CD43 (leukosialin), CD44 (H-CAM), CD54 (ICAM-1), embryonal N-CAM (e-NCAM), and EMA (episialin). In all cases, the surfaces of IL aggregates reacted for CD44 but were consistently negative for CD29; also absent was CD54. Conversely, the integrity of the endothelial cells was underscored by their even reactivity for CD29, CD44, and CD54. Given that CD29 is currently regarded as critical for lymphocyte trafficking in general and for transvascular migration in particular, and CD54 is also involved in transvascular lymphocyte migration, we conclude that their consistent absence in IL may contribute to its intravascular and disseminated distribution pattern. The rather frequent association of IL with various conventional lymphomas is known; yet, one of our cases appears to be the first report of IL associated with a low-grade MALT lymphoma. PMID- 10685638 TI - Second primary epithelial malignancy of nasopharynx and nasal cavity after successful curative radiation therapy of nasopharyngeal carcinoma. AB - Patients with head and neck cancer are at high risk of developing additional second primary tumors in the aerodigestive tract as a result of the field cancerization phenomenon. In this context, the appearance of a new neoplasm often poses a problem in differential diagnosis between recurrence and new primary tumor. Twelve patients with nasopharyngeal carcinoma (NPC) who received radiation therapy for the primary tumor and developed a second epithelial malignancy in the nasal cavity or nasopharynx during the follow-up period are presented in this report. The differentiation between the 2 entities based on the spatiotemporal relations, histological features, and the status of Epstein-Barr virus in tumor lesions are also presented. Our study showed that the epithelial malignancy after NPC having late-onset or prolonged interval (range, 5 to 18 years), different histological patterns (keratinizing squamous cell carcinoma, neuroendocrine carcinoma, or small cell carcinoma) distinct from the primary NPC (differentiated or undifferentiated nonkeratinizing carcinoma), and absence of Epstein-Barr virus, indicate a newly developed tumor rather than recurrent NPC. Our observations showed for the first time that second primary epithelial malignancy developed in the nasal cavity or nasopharynx years after curative therapy for NPC with a prevalence of 0.4% (12/2,794). Wild-type p53 protein was expressed more often in the original NPC (9 of 12) than in the second tumors (4 of 10), but the significance was not statistically significant (P = .2048). Genomic analysis for p53 mutation and in situ hybridization for human papillomavirus showed negative results, indicating that both important molecular events in NPC or head and neck cancer play a small role in this particular type of newly developed second malignant tumor. More studies are warranted for further clarification for the development of second epithelial malignancies in treated NPC patients. PMID- 10685639 TI - Loss of heterozygosity in P53, BRCA1, and estrogen receptor genes and correlation to expression of p53 protein in ovarian epithelial tumors of different cell types and biological behavior. AB - Loss of heterozygosity (LOH) of tumor suppressor genes (TSGs) in ovarian epithelial tumors of differing cell types and biological behavior has not been thoroughly investigated. Moreover, there have been conflicting reports correlating LOH of the p53 gene to overexpression of p53 protein. This study evaluated 34 formalin-fixed, paraffin-embedded ovarian epithelial tumors for LOH by polymerase chain reaction (PCR) for the following microsatellite markers: TP53(17p13.1/p53 gene), D17S579(17q/BRCA1 gene), and ESR (6q24-27/estrogen receptor gene). LOH of the TP53 marker was detected in 4 (44%) of 9 informative serous cystadenocarcinomas (SCa) but in 0 of 4 informative clear cell carcinomas (CCa) and 0 of 5 informative serous tumors of low malignant potential (SLMP). LOH of the BRCA1 marker was detected in 5 (83%) of 6 informative SCa, but in 1 (13%) of 8 informative CCa and 1 (14%) of 7 informative SLMP. LOH of the ESR marker was detected in 4 (50%) of 8 informative SCa, but in 0 of 4 informative CCa and 1 (16%) of 6 informative SLMP. p53 protein overexpression was present in 8 of 12 SCa but did not correlate to TP53 LOH. LOH for TP53, D17S579/ BRCA1, and ESR is common in ovarian SCa, and is observed in primary tumors as well as metastases. In contrast, these genetic alterations are less common in CCa and in the biologically less aggressive SLMP tumors. These data suggest different mechanisms of oncogenesis in ovarian epithelial tumors of different cell types and biological behavior. PMID- 10685640 TI - Reactive vascular lesion of nasal septum simulating angiosarcoma in a cocaine abuser. AB - We report the case of an exuberant ulcerative angiomatoid nasal lesion in a cocaine abuser. The lesion was made up of polymorphous endothelial cells with occasional mitoses, arranged in a lobular pattern with infiltrative-looking areas. There were extensive areas of thrombosis with focal recanalization. Intravascular proliferation was not observed. The clinical, radiological, and histological features suggested hemangiosarcoma as the main differential diagnosis, but the lobular architecture of the lesion and the widespread thrombosis favoured the diagnosis of a benign reactive process. PMID- 10685641 TI - The ciliated hepatic foregut cyst, an unusual bronchiolar foregut malformation: a histological, histochemical, and immunohistochemical study of 7 cases. AB - The ciliated hepatic foregut cyst is an unusual solitary cystic lesion of the liver. In a series of 7 cases of hepatic ciliated cysts, we performed a histological, histochemical, and immunohistochemical study to better define the histogenesis of this rare entity. The patients were 4 women and 3 men, aged 39 to 75 years. Four patients presented with abdominal pain. In 3 cases the cyst was discovered incidentally on ultrasonography. The cysts measured from 1 to 4 cm in diameter. Microscopically, the lining of the columnar epithelium was composed of ciliated cells and mucin secreting goblet cells. The wall was composed of bands of smooth-muscle fibers surrounded by an outer fibrous capsule. The goblet cells stained with PAS, alcian blue, and high-iron diamine. The immunohistochemical study showed that endocrine cells were present within the cyst epithelium, positive for chromogranin, synaptophysin, bombesin, and calcitonin, and negative for serotonin, somatostatin, glucagon, insulin, gastrin, and pancreatic polypeptide. In all the cases, immunoreactivity of some cells for CC10 strongly suggested the presence of Clara cells. Our study shows that the epithelium lining ciliated hepatic foregut cysts has histological, histochemical, and immunohistochemical features similar to those observed in the bronchiolar epithelium. This lesion is a developmental ventral foregut abnormality that could arise from a bronchiolar bud of the tracheobronchial diverticulum. PMID- 10685642 TI - Molecularly confirmed primary prostatic synovial sarcoma. AB - We report the second molecularly-confirmed primary prostatic synovial sarcoma. The diagnosis was particularly elusive at the light microscopic level in that the tumor failed to show epithelial differentiation, but it did show combined spindle cell and poorly differentiated (round-cell) morphologies. The immunohistochemical staining profile was nonspecific and potentially misleading. Only by demonstration of the characteristic SYT-SSX gene fusion of synovial sarcoma by reverse transcriptase polymerase chain reaction (RT-PCR) analysis of RNA extracted from archival material could the diagnosis be confirmed. This case further illustrates the use of this technique in diagnostic pathology. PMID- 10685643 TI - Subacute hepatic failure associated with a new antidiabetic agent, troglitazone: a case report with autopsy examination. AB - An autopsy case of fatal subacute hepatic failure after administration of troglitazone is described. The liver dysfunction developed about five months after the patient, a sixty-three-year-old woman, had been initially treated with troglitazone. The patient developed hepatic failure and died despite various hepatic auxiliary treatments such as plasmapheresis. Autopsy findings revealed focal liver cell necrosis, cholestasis and steatosis with infiltration of lymphocytes and neutrophils and lack of regenerative activity. The causative mechanism of liver dysfunction may be metabolite aberration, as a result of accumulation of hepatotoxic metabolite(s), in a category of idiosyncratic liver injury. It is proposed to monitor liver function strictly and periodically for the diabetic patients prescribed troglitazone. PMID- 10685644 TI - Epstein-Barr virus-associated Hodgkin's lymphoma and legionella pneumophila infection complicating treatment of juvenile rheumatoid arthritis with methotrexate and cyclosporine A. AB - We describe the case of a 53-month-old girl with juvenile rheumatoid arthritis (JRA), complicated by the occurrence of Hodgkin's lymphoma and Legionella pneumophila infection during immunosuppressive treatment with methotrexate (MTX) and cyclosporine A (CSA). The girl had received variable anti-inflammatory combination therapy, including MTX for 28 months and CSA for 3 months. Thirty-six months after the onset of arthritis, the girl presented with an enlargement of the lymph nodes of the mediastinum, the hilum of the lungs, and the abdomen. Concomitantly, a diagnosis of Legionella pneumonia was rendered. Autopsy showed Epstein-Barr virus (EBV)-associated nodular sclerosing Hodgkin's lymphoma. The neoplastic cells were positive for CD15, CD 30, and latent membrane protein 1 (LMP 1). The present case is the second reported to occur in a child, and it lends support to the hypothesis that immunosuppressive treatment may contribute to an increased risk of the development of EBV-associated lymphoproliferative disorders (LPD) in pediatric patients suffering from JRA. PMID- 10685645 TI - Low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) of thymus. AB - This report describes a low-grade B-cell lymphoma of mucosa associated lymphoid tissue (MALT) involving the thymus of a 63-year-old woman with features suggestive of a connective tissue disease. Sections of the thymic lesion and of a lung biopsy performed at the same operation were examined histologically and by immunohistochemistry using the monoclonal antibodies CD45, CD20, CD79a, CD3, CD45RO, and AE1/AE3. Polymerase chain reaction (PCR) for immunoglobulin heavy chain gene rearrangement was also performed. The dense infiltrate of small lymphoid cells intimately admixed with ramifying epithelial elements, some of which had undergone cystic change, closely resembled a thymoma. The lymphoid infiltrate comprised centrocyte-like cells, small lymphocytes, plasma cells, and blasts. Most of the lymphoid cells were immunoreactive with the B-cell markers CD20 and CD79a, and PCR showed clonal immunoglobulin heavy chain gene rearrangement. The lung biopsy showed dense infiltration by small lymphoid cells, morphologically suggestive of lymphoid interstitial pneumonia. However, PCR showed a weak band in the amplification for immunoglobulin heavy chain gene rearrangement, identical to that within the thymus and suggesting either recirculation of cells to accumulated MALT or subhistological lymphoma. MALT lymphoma may rarely involve the thymus, and pathologists should be aware of this to avoid misdiagnosis as a thymoma. Immunohistochemical and/or molecular studies are of value in this regard. MALT lymphomas of the thymus, common with those arising in other organs, may develop in the setting of a connective tissue disease. PMID- 10685646 TI - Congenital primitive epithelial tumor of the liver showing focal rhabdoid features, placental involvement, and clinical features mimicking multifocal hemangioma or stage 4S neuroblastoma. AB - We describe an unusual case of congenital primitive epithelial tumor of the liver with focal rhabdoid features. The present case is unique and informative in the following aspects: (1) a first case of congenital epithelial tumor of the liver with no hepatocytic differentiation but focal rhabdoid features, (2) clinical similarities to multicentric hemangioma or stage 4S neuroblastoma, (3) diagnosis obtained from histological examination of the placenta immediately after birth. PMID- 10685647 TI - Eradication of Borrelia burgdorferi infection in primary marginal zone B-cell lymphoma of the skin. AB - Primary cutaneous B-cell lymphomas have been associated with Borrelia burgdorferi, the spirochete responsible for Lyme disease. Recently, cutaneous marginal zone B-cell lymphoma has been proposed as a distinct clinical pathological entity. We report a case of primary cutaneous marginal zone lymphoma, associated with B burgdorferi infection. Polymerase chain reaction (PCR) amplification of the third complementarity determining region (CDR3) of the immunoglobulin heavy chain gene showed the presence of a monoclonal lymphoproliferation, therefore strengthening the histological diagnosis of a malignant process. B burgdorfer-specific hbb gene sequences were detected by PCR in the lymphoma tissue at diagnosis but not after antibiotic treatment. A nearly complete clinical and histological regression was observed after B burgdorferi eradication, with immunohistochemistry studies showing disappearance of plasma cell differentiation and a marked decline in the number of CD3+ T cells and Ki 67+ cells. Our case confirms the link between B burgdorferi and some cutaneous lymphomas. The disappearance of the microorganism accompanied by the unequivocal decrease of most indicators of active T- and B-cell immune response strongly supported a pathogenetic role for B burgdorferi in sustaining an antigen-driven development and growth of this cutaneous marginal zone lymphoma. Antibiotic therapy (analogous to Helicobacter pylori infection in gastric MALT lymphoma) might be helpful with the aim of averting or at least deferring the indication for more aggressive treatment. PMID- 10685648 TI - Molecular evidence of field cancerization in a patient with 7 tumors of the aerodigestive tract. AB - Exposure of the mucosa of the upper aerodigestive tract to carcinogens can induce genetic changes resulting in various independent clones of neoplastic growth, a concept defined as "field cancerization." The risk of developing multiple tumors in this compartment of the body is well established. We studied 6 distinct tumors of the upper aerodigestive tract of a single patient for loss of heterozygosity (LOH), microsatellite instability (MSI), p53 mutations, and K-ras codon 12 point mutations. We detected a unique pattern of LOH and p53 mutations in all 6 tumors. No tumor showed a K-ras mutation or MSI. The results support the mechanism of "field cancerization" and illustrate the potential power of molecular techniques to elucidate pathogenesis. PMID- 10685649 TI - Primary signet-ring cell carcinomas of the lung. PMID- 10685650 TI - Interleukin-2 and the development of immunotherapy for the treatment of patients with cancer. PMID- 10685651 TI - Interleukin-2 in metastatic melanoma: what is the current role? PMID- 10685652 TI - High-dose recombinant interleukin-2 therapy in patients with metastatic melanoma: long-term survival update. AB - PURPOSE: To update response duration and survival data for patients with metastatic melanoma receiving the high-dose IV bolus recombinant interleukin (IL) 2 regimen. PATIENTS AND METHODS: Two hundred seventy assessable patients were entered into eight clinical trials conducted between 1985 and 1993. IL-2 600,000 or 720,000 IU/kg was administered by 15-minute intravenous infusion every 8 hours for up to 14 consecutive doses over 5 days as clinically tolerated with maximum support, including pressors. A second, identical cycle of treatment was scheduled following 6 to 9 days of rest, and courses could be repeated every 6 to 12 weeks in stable or responding patients. Responding patients received up to five courses (two cycles/course) of treatment. All data were updated through December 1998 using report forms completed by the clinical investigators. RESULTS: The objective overall response rate was unchanged from the previous report. Tumor responses were seen in 16% of patients, with complete responses in 17 (6%) and partial responses in 26 (10%). Median survival for the group as a whole is now 12 months. Median follow-up time for surviving patients exceeds 7 years. Median duration of response for the 43 responding patients and the 26 patients with partial responses remained unchanged at 8.9 and 5.9 months, respectively. Response durations ranged from 1.5 to > 122 months. The median duration of complete responses has yet to be reached, but is at least 59 months. Thirty-one patients (11%) were alive as of last contact; 28 were confirmed, including 18 responding patients. Three patients were lost to follow-up at > 1, > 13, and > 104 months. Twelve responding patients remained continually disease- or progression-free from > 70 to > 150 months following initiation of therapy. Disease progression was not observed in any patient who was responding as of the last report or in any patient responding for longer than 30 months. CONCLUSION: These data continue to support the notion that high-dose IL-2 produces durable responses in some patients with metastatic melanoma and should be considered a therapeutic option for appropriately selected patients with this disease. PMID- 10685653 TI - Rationale for intergroup trial E-3695 comparing concurrent biochemotherapy with cisplatin, vinblastine, and DTIC alone in patients with metastatic melanoma. AB - PURPOSE: The modest activity of chemotherapy and biologic agents in the treatment of advanced metastatic melanoma has prompted investigators to consider combinations of chemotherapy and biologic agents (i.e., biochemotherapy) as a way of improving response rates and survival. Although biochemotherapy has generated a great deal of interest over the last several years, and these regimens have produced high response rates in single-institution phase II trials, they have yet to demonstrate a significant survival benefit in randomized trials compared with either chemotherapy or biotherapy alone. METHODS: The available literature regarding the clinical experience with single- and multiagent chemotherapy, immunotherapy, and biochemotherapy was reviewed. RESULTS: Treatment of metastatic melanoma with either single-agent or combination chemotherapy regimens is clearly suboptimal; the majority of responses are partial and of short duration. In contrast, interleukin (IL)-2 produces long-term durable complete remission in a subset of patients. Over 1,000 patients have been treated with IL-2-based biochemotherapy regimens in single-institution phase II trials, and response rates have ranged from 40% to 60%. Most encouraging have been the durable responses observed in 10% to 20% of patients in most of these trials. Large databases, including two meta-analyses, have confirmed the substantial improvement in response rate associated with biochemotherapy regimens that include both IL-2 and interferon alfa (IFN-alpha) compared with chemotherapy or biotherapy alone. Biochemotherapy is currently being evaluated in randomized controlled trials to determine if this treatment strategy can provide a survival benefit compared with current standard treatments. A pilot study at Beth Israel Deaconess Medical Center has demonstrated the feasibility of administering a modification of a concurrent biochemotherapy regimen, initially described by Legha et al, consisting of cisplatin, vinblastine, and dacarbazine (CVD) plus IL 2 and IFN-alpha in the cooperative group setting. CONCLUSION: These studies provided the rationale for intergroup trial E-3695, which is currently randomizing patients to concurrent biochemotherapy with CVD plus IL-2 and IFN alpha versus CVD alone. PMID- 10685654 TI - Strategies to reduce side effects of interleukin-2: evaluation of the antihypotensive agent NG-monomethyl-L-arginine. AB - PURPOSE: The clinical utility of high-dose intravenous recombinant interleukin (IL)-2 therapy is limited by severe toxicity including hypotension, fever, chills, pulmonary edema, and oliguria Hypotension has been previously shown to result from excessive vascular relaxation due to overproduction of the endogenous vasodilator nitric oxide. Nitric oxide production can be decreased by administration of the competitive enzyme inhibitor NG-monomethyl-L-arginine (NMA). A clinical trial to investigate the dose-dependent effects of NMA on blood pressure was undertaken in patients with metastatic renal cell carcinoma. PATIENTS AND METHODS: Patients with metastatic renal cell carcinoma receiving a 5 day continuous infusion of IL-2 (18 million IU/m2/d) who developed hypotension were treated with increasing doses of NMA, ranging from 3 to 36 mg/kg. RESULTS: Twenty-three patients received a total of 61 courses of IL-2; 18 of these patients developed hypotension and received NMA. Antihypotensive activity was observed at all dose levels, and the duration of the effect varied directly with the dose of NMA. At the higher dose levels tested (12 to 36 mg/kg), increased pulmonary vascular resistance and decreased cardiac output were observed. Patients experiencing a significant decrease in cardiac output received dobutamine (2.5 to 10 microg/kg/min). Pulmonary capillary wedge pressure was unaffected by administration of NMA. One patient treated at 24 mg/kg (bolus) experienced a major motor seizure, but no neurologic disorders were observed in other patients treated with NMA doses of 24 to 36 mg/kg. No other adverse events involving hepatic, renal, or hematologic systems were attributed to NMA. Three patients received NMA by an initial bolus followed by a continuous infusion. Similar antihypotensive effects were noted, and these patients were able to complete a full 5-day course of IL-2. CONCLUSION: The antihypotensive effects of NMA appear to be optimal at a dose of 24 mg/kg, with maintenance doses of 8 mg/kg every 4 to 6 hours. At this dose level, blood pressure was restored, and IL-2 associated vasodilatation was fully reversed. Coincident with the reversal of hypotension, the state of high cardiac output was also reversed by NMA administration. These results suggest that NMA may be effective for alleviating the hypotensive effects of high-dose IL-2 therapy in cancer patients. PMID- 10685655 TI - Interleukin-2 in the treatment of hematologic malignancies. PMID- 10685656 TI - Immunotherapy with interleukin-2 after hematopoietic cell transplantation for hematologic malignancy. AB - PURPOSE: The results of trials using interleukin (IL)-2-based therapy in leukemia and after hematopoietic stem cell transplant suggest that such therapy could have an impact on preventing disease relapse in patients with hematologic malignancy who achieve a minimal disease state. The use of immunotherapy in the autologous transplant setting is modeled in part on the well-characterized immunotherapeutic effect of the graft-versus-tumor response in patients undergoing allogeneic transplantation. The graft-versus-tumor response, mediated by donor cells, contributes to the higher cure rates seen in patients undergoing allogeneic transplant for the treatment of a variety of hematologic malignancies, including acute and chronic myelogenous and lymphoblastic leukemia, myeloma, and lymphoma PATIENTS AND METHODS: The literature was reviewed, and we relate our own clinical experience with IL-2 therapy in this setting. RESULTS: Preclinical in vitro and animal data show a variety of leukemia cells are sensitive to autologous IL-2 activated effector cells. In addition, laboratory studies show that IL-2 can be used to activate antitumor cellular responses from bone marrow and peripheral blood without compromising hematopoiesis. Most importantly, in vitro studies show that chemoresistant malignant hematopoietic cells are sensitive to IL-2-induced cell death, thus emphasizing the lack of cross resistance to immunologic-based therapeutics. The results of phase I and II studies conducted in patients with acute myelogenous leukemia in first or subsequent remission suggest that autologous IL-2-activated cells may mediate an antitumor response and aid in preventing relapse after autologous transplantation. Clinical trials to determine the role of IL-2 after transplantation for the treatment of acute and chronic myelogenous leukemia, multiple myeloma, and lymphoma are ongoing. CONCLUSION: These studies will help define the optimal dose and schedule of IL-2 and its role in augmenting therapeutic immune-mediated autologous responses. PMID- 10685657 TI - Children's cancer group trials of interleukin-2 therapy to prevent relapse of acute myelogenous leukemia. AB - PURPOSE: Up to 80% of children with acute myelogenous leukemia treated with intensive chemotherapy achieve remission; however, a large proportion of patients develops recurrent disease. Because interleukin (IL)-2 can induce remission in patients with overt evidence of acute myelogenous leukemia, we hypothesized that it might prevent relapse when administered to patients in first remission after intensive consolidation chemotherapy. A pilot Children's Cancer Group (CCG) trial (CCG-0941) demonstrated the feasibility of this approach, and we initiated a prospective randomized trial (CCG-2961) to further evaluate the safety and potential efficacy of IL-2 therapy in preventing relapse of acute myelogenous leukemia. PATIENTS AND METHODS: In trial CCG-0941, 21 pediatric patients in complete remission following induction and consolidation chemotherapy on protocol CCG-2941 received IL-2 therapy. In CCG-2961, 79 patients in complete remission were randomized as of February 1999 to receive either IL-2 (n = 39) or no further therapy. In both trials, recombinant IL-2 was given at a dose of 9 million IU/m2/d by continuous intravenous infusion for 4 days. After 4 days of rest, IL-2 was resumed at a dose of 1.6 million IU/m2/d for 10 days by continuous infusion. We monitored patients for toxicity and relapse. RESULTS: The majority of patients treated with IL-2 in these two trials experienced some degree of fever. Seven of 60 patients (12%) had clinically significant rashes, and grade 3 vascular leak syndrome and hypotension have each been observed in five patients (8%). Hypotension resolved promptly after treatment with intravenous fluids. No patients have experienced renal toxicity or required cardiac vasopressors or transfer to an intensive care unit; there have been no treatment-related deaths. Overall, the incidence and severity of adverse events remain similar in the two trials. Total projected accrual to the IL-2 randomization is anticipated to be 326 patients, and relapse and survival data remain blinded. CONCLUSION: The dose and schedule of IL-2 used in these two trials continue to be reasonably well tolerated by children with acute myelogenous leukemia in first remission. Any conclusions with regard to efficacy must await completion of the randomized trial. PMID- 10685658 TI - Cytokine replacement in patients with HIV-1 non-Hodgkin's lymphoma: the rationale for low-dose interleukin-2 therapy. AB - PURPOSE: The drastic increase in the incidence of non-Hodgkin's lymphoma in patients infected with HIV-1 is testimony to the fact that our immune system is critical for the prevention of certain malignancies. Preclinical and clinical studies were conducted to (1) gain further insight into defects in immunity that can lead to malignant transformation and (2) determine if certain immune deficiencies could be corrected by cytokines delivered at doses that result in near-physiologic concentrations in vivo. METHODS: We have used the severe combined immune deficient mouse engrafted with human peripheral blood leukocytes from healthy individuals who are seropositive for the Epstein-Barr virus to study the spontaneous development of malignant Epstein-Barr virus-positive human B-cell lymphoproliferative disorder. RESULTS: We have demonstrated in this model that, in the absence of CD4+ T cells, cytokine replacement with low-dose interleukin (IL)-2 therapy can prevent Epstein-Barr virus-positive human B-cell lymphoproliferative disorder by interacting with mouse natural killer and human CD8+ T cells. We review our clinical experience with administration of low-dose IL-2 therapy in patients with HIV-1-related cancer, noting minimal toxicity and significant immune modulation. We provide evidence that this therapy can favorably alter the type 1 cytokine profile in vivo in these patients, and improve the cellular response to infectious insults in vitro. CONCLUSION: Early clinical studies with low-dose IL-2 therapy in patients with HIV-1-related lymphoma suggest that this therapy may have a role in the prevention and treatment of this disease. PMID- 10685659 TI - Renal cell carcinoma: current status and future plans. PMID- 10685660 TI - Long-term survival update for high-dose recombinant interleukin-2 in patients with renal cell carcinoma. AB - PURPOSE: To update response duration and survival data for patients with metastatic renal cell carcinoma treated with high-dose interleukin (IL)-2. PATIENTS AND METHODS: Two hundred fifty-five assessable patients were entered onto seven phase II clinical trials. Recombinant IL-2 600,000 or 720,000 IU/kg was administered by 15-minute intravenous infusion every 8 hours for up to 14 consecutive doses over 5 days as clinically tolerated with maximal support. A second, identical cycle of treatment was scheduled following 5 to 9 days of rest, and courses could be repeated every 6 to 12 weeks in stable or responding patients. All data were updated as of December 1998, with report forms completed by the clinical investigators. These data had last been updated as part of the Food and Drug Administration reporting requirements in 1996. RESULTS: Objective responses previously have been reported in 37 of 255 patients (15%) with 17 complete responses (7%) and 20 partial responses (8%). These data remain unchanged from previous reports. Median response duration for all objective responders remains unchanged at 54 months, but the range now extends from 3 to > 131 months. Median duration for all complete responses has not yet been reached, but was at least 80 months (range, 7- > 131 mo) at the time of this analysis. Median duration for all partial responses remains 20 months (range, 3- > 126 mo). Median survival time for all 255 patients remains 16.3 months, with 10% to 20% of patients estimated to be alive 5 to 10 years after treatment with high-dose IL-2. CONCLUSION: With prolonged follow-up, treatment with high-dose recombinant IL-2 remains extremely effective for a subset of patients with metastatic renal cell carcinoma. PMID- 10685661 TI - The future role of interleukin-2 in cancer therapy. PMID- 10685662 TI - Interleukin-2: developing additional cytokine gene therapies using fibroblasts or dendritic cells to enhance tumor immunity. AB - PURPOSE: Recombinant interleukin (IL)-2 administration can mediate regression of solid tumors in patients with melanoma and renal cell carcinoma. A better understanding of the mechanisms of IL-2-mediated antitumor effects has led to the investigation of novel immunotherapeutic approaches. The rationale for these immunotherapeutic approaches and the results of preliminary clinical studies are presented. PATIENTS AND METHODS: The therapeutic potential of dendritic cells and the role of FLT3 ligand, a potent hematopoietic growth factor, was investigated in a variety of preclinical models. In addition, a clinical study with autologous dendritic cells pulsed with synthetic melanoma peptides derived from the MART1/ Melan A, gp100, and tyrosinase proteins was conducted. Twenty-eight human leukocyte antigen (HLA)-A2+ melanoma patients received an average of 106 dendritic cells a week for 4 weeks. RESULTS: In a murine liver metastases model, FLT3 ligand administration alone or in combination with IL-12 or IL-2 had significant antitumor effects and resulted in significant infiltration of the tumor border by lymphocytes and dendritic cells, which was associated with an increased number of apoptotic figures. Administration of melanoma peptide-pulsed dendritic cells to 28 patients with advanced metastatic melanoma produced a complete response in two patients and a partial response in one. Significant infiltration of T cells and dendritic cells into melanoma lesions was observed. CONCLUSION: These studies confirm the feasibility of immunotherapeutic approaches using dendritic cells and FLT3 ligand and demonstrate their potential antitumor activity. These approaches may be effective for patients with metastatic melanoma and other solid tumors and will likely be used to improve the efficacy of IL-2 based immunotherapy. PMID- 10685663 TI - Potentiation of immunologic responsiveness to dendritic cell-based tumor vaccines by recombinant interleukin-2. AB - PURPOSE: Dendritic cells (DC) can elicit potent immune responses to tumors through their capacity to efficiently process and present tumor-associated antigens. In a variety of animal tumor models, vaccines based on tumor lysate pulsed DC (TP-DC) have been shown to effectively immunize against lethal tumor challenges as well as to treat established growing tumors at skin and organ sites. The antitumor effects elicited by TP-DC-based vaccines in vivo have been shown to be mediated by tumor-specific proliferative, cytotoxic, and cytokine secreting host-derived T cells. Because of the critical involvement of T cells in the antitumor immune response, we have been investigating whether the systemic administration of recombinant interleukin (IL)-2 can enhance the therapeutic efficacy of TP-DC-based tumor vaccines. MATERIALS AND METHODS: Immunization with TP-DC plus IL-2 administration was evaluated to determine if this combination could enhance protective immunity toward a weakly immunogenic sarcoma (MCA-207) and a poorly immunogenic subline (D5) of the B16 melanoma and mediate therapeutic rejection of established tumors in C57BL/6 (B6) mouse models. RESULTS: We have demonstrated in our murine models that the addition of IL-2 at relatively nontoxic doses can markedly augment the antitumor activity of TP-DC-based tumor vaccine therapies against both a weakly immunogenic sarcoma and a poorly immunogenic melanoma. Animals treated with the combination exhibited significantly greater protection from tumor-cell challenge, significantly greater regression of established tumors, and significantly longer mean survival time than with either TP-DC or IL-2 therapy alone. The mechanism operative in vivo appears to involve the enhancement of immune T-cell function. CONCLUSION: These preclinical studies demonstrate the potential of this novel treatment strategy and support the rationale for planned phase I/II clinical trials of TP-DC-based vaccines plus IL-2 in patients with advanced melanoma and colorectal cancer. PMID- 10685664 TI - The future of interleukin-2: enhancing therapeutic anticancer vaccines. AB - PURPOSE: The purpose of our efforts is to trigger the immune destruction of established cancer. Interleukin (IL)-2 can mediate the regression of tumors in patients with melanoma and renal cell carcinoma. In animal models, the antitumor effects of IL-2 are mediated by T lymphocytes. Stimulation with specific antigen can enhance the ability of T cells to respond to IL-2 by triggering the rapid upregulation of the high-affinity IL-2 receptor. We are seeking to design recombinant and synthetic vaccines capable of preferentially priming T cells with specificity for tumor cells. METHODS: The antitumor activity of experimental vaccines is being studied preclinically using recently developed murine models that employ the mouse homologues of human tumor-associated antigens. Once the most effective experimental vaccines are optimized in experimental animals, clinical trials can be conducted. Vaccines are being evaluated for their ability to mediate the regression of established tumors, and a variety of immunologic correlates are being measured. RESULTS: In animal models, vaccines based on molecularly defined tumor-associated antigens expressed in viral vectors or delivered as "naked" DNA stimulate the expansion of CD4+ and CD8+ tumor-specific T lymphocytes. Coadministration of IL-2 with these vaccines dramatically enhances their ability to mediate the regression of established cancer. In the clinic, treatment of melanoma patients with peptide vaccine and IL-2 resulted in objective responses in approximately 40% of patients, a response rate more than twice that typically achieved with IL-2 alone. Paradoxically, tumor-specific CD8+ T-cell levels were not increased in these patients. CONCLUSION: The addition of recombinant and synthetic cancer vaccines to a regimen of IL-2 can result in improved antitumor responses in both animal models and melanoma patients. Vaccine primed, tumor-specific T cells may preferentially proliferate upon administration of IL-2. The apparent lack of increase in CD8+ T-cell numbers in this setting suggests that the vaccine-primed T cells functionally disappear after a transient period of activation. Preventing the disappearance of activated T cells upon IL-2 administration-for example, by blocking proapoptotic signals-may enhance the therapeutic effectiveness of anticancer vaccines. PMID- 10685665 TI - Tumor-induced dysfunction in interleukin-2 production and interleukin-2 receptor signaling: a mechanism of immune escape. AB - PURPOSE: The development of an effective antitumor immune response is compromised in patients with renal cell carcinoma. Despite significant infiltration by T lymphocytes into renal tumors, no detectable induction of gene expression is associated with the generation of an antitumor immune response. Tumor-induced down-regulation of interleukin (IL)-2 expression may contribute to the impaired development of the T cell-mediated antitumor immune response. Within renal tumors, there is no detectable expression of IL-2 or the IL-2 receptor alpha chain, and only low levels of interferon gamma (IFN-gamma) mRNA are detected. Products in the tumor environment may suppress the expression of these genes, thus inhibiting production of type 1 helper T cell cytokines. METHODS: Peripheral blood lymphocytes obtained from healthy volunteers were exposed to supernatants from renal cell carcinoma explants, and the immunologic consequences of this were assessed using a variety of molecular assays. RESULTS: Soluble products from renal tumor explants can inhibit the production of IL-2 and IFN-gamma by peripheral blood lymphocytes and can suppress T-cell proliferation. Soluble products from renal cell carcinoma explants appear to block the nuclear translocation of nuclear factor kappa B (NFkappaB) proteins p50 and RelA without affecting cytoplasmic levels of these proteins. In some experiments, a reduction in the nuclear translocation of other transcription factors involved in IL-2 gene expression, including nuclear factor of activated T cells and accessory protein 1, was observed. Gangliosides isolated from tumor supernatants blocked the production of IL-2 and IFN-gamma in response to ionomycin plus phorbol myristate acetate stimulation. These gangliosides also inhibited stimulus-dependent activation and nuclear accumulation of NFkappaB. Coculture experiments demonstrated that renal cell carcinoma lines known to express gangliosides could inhibit the activation of NFkappaB in normal T cells and the Jurkat T-cell line. Supernatants from renal cell carcinoma explants and renal cell carcinoma cell lines can also suppress the proliferation of normal T cells, thus reproducing another defect observed in tumor-infiltrating lymphocytes. Supernatants from renal cell carcinoma tumors also appear to inhibit signaling through the IL-2 receptor. Although tumor supernatants had little effect on IL-2 receptor (alpha, beta or gamma) expression, they did block expression of JAK3, a key kinase involved in signaling through the IL-2 receptor pathway. Moreover, downstream events in IL-2 receptor signaling linked to JAK3 were impaired in T cells treated with tumor supernatants. CONCLUSION: These findings suggest that soluble products from renal tumors may suppress T-cell responses by blocking both IL-2 production and normal IL-2 receptor signaling. PMID- 10685666 TI - Expanding the indications for surgery and adjuvant interleukin-2-based immunotherapy in patients with advanced renal cell carcinoma. AB - PURPOSE: To determine the role of surgery and adjuvant interleukin (IL)-2-based immunotherapy in the treatment of patients with advanced metastatic renal cell carcinoma PATIENTS AND METHODS: The survival of 354 consecutive patients with metastatic renal cell carcinoma treated with IL-2-based immunotherapy through the UCLA Medical Center Kidney Cancer Program was analyzed There were five groups of patients. Patients who initially presented with metastatic disease received either (1) IL-2 therapy with primary tumor in place; (2) nephrectomy followed by IL-2 therapy, or (3) nephrectomy followed by immunotherapy with IL-2 plus tumor infiltrating lymphocytes. Patients who underwent nephrectomy for localized disease were divided into two groups: (4) those who developed metastatic disease > or = 6 months after nephrectomy and then received IL-2 therapy; and (5) those who developed metastatic disease < 6 months after nephrectomy and then received IL-2 therapy. Kaplan-Meier survival curves were generated for all patient groups. RESULTS: Among patients who received IL-2-based immunotherapy with their primary tumor in place (group 1; n = 36), 1- and 2-year survival rates were 29% and 4%, respectively, compared with 1- and 2-year survival rates of 67% and 44%, respectively, for all similar patients who underwent nephrectomy prior to IL-2 therapy (n = 235). Among patients initially presenting with metastatic disease who underwent nephrectomy followed by IL-2 therapy without tumor-infiltrating lymphocytes (group 2; n = 69), the 1- and 2-year survival rates were 53% and 25%, respectively. The best survival was observed in patients treated with nephrectomy followed by IL-2 plus tumor-infiltrating lymphocyte therapy (group 3; n = 102), which yielded 1- and 2-year survival rates of 73% and 55%, respectively. Among patients initially undergoing nephrectomy for localized disease, patients receiving IL-2-based therapy for subsequent metastasis > or = 6 months following nephrectomy (group 4; n = 128) had 1- and 2-year survival rates of 64% and 40%, respectively, compared with 45% and 15%, respectively, for patients developing metastasis < 6 months after nephrectomy (group 5; n = 19). CONCLUSION: The role of surgery prior to IL-2-based immunotherapy remains controversial Our data demonstrate that aggressive surgery is safe, causing minimal morbidity despite extensive tumor involvement, and significantly improves survival outcomes in patients with metastatic renal cell carcinoma when carried out in conjunction with an IL2-based immunotherapy regimen. PMID- 10685667 TI - Long-term follow-up of patients with metastatic renal cell carcinoma treated with intravenous recombinant interleukin-2 in Europe. AB - PURPOSE: The median survival for patients with metastatic renal cell carcinoma (mRCC) is generally < 1 year. Immunotherapy with high-dose recombinant interleukin (IL)-2 has been reported to produce objective responses in approximately 15% of treated patients and is associated with durable complete responses and prolonged survival in responding patients. The impact of IL-2 therapy on survival of metastatic renal cell carcinoma patients has begun to emerge, based on long-term follow-up data from large databases. Combinations of IL-2 and interferon alfa (IFN-alpha) have also been intensively investigated in mRCC. PATIENTS AND METHODS: Between 1987 and 1990, 281 mRCC patients were treated with continuous infusion IL-2 in three European multinational, single-arm phase II trials. Long-term treatment outcomes for these patients were analyzed, and the results are presented here. The results of a large, randomized French cooperative group trial (the Cancer Renal Cytokine [CRECY] study) that enrolled 425 patients between 1991 and 1995 are also summarized. Patients on this trial were randomized to treatment with IL-2 alone, IFN-alpha alone, or the combination. RESULTS: Among patients included in the 281-patient database, the objective response rate was 15%. Median survival was 10 months; 41% of patients were alive at 1 year, 22% were alive at 2 years, and 8% were alive at 5 years. Among patients with a complete or partial response, 60% and 18% were alive at 5 years, respectively. No clinical factors were predictive for response or survival; however, no patient with a high endogenous IL-6 level at diagnosis responded to IL-2 therapy. The CRECY trial demonstrated that the combination of IL-2 and IFN-alpha induced a significantly higher response rate (P < 0.01) and significantly improved 1-year event-free survival (P = 0.01) compared with either agent alone, but overall survival was not significantly different between the three treatment groups. CONCLUSION: The European experience suggests that the 5-year survival rate for metastatic renal cell carcinoma patients treated with high-dose continuous infusion IL-2 therapy is approximately 8% and that the majority of the therapeutic benefit is restricted to patients achieving a complete response. Therefore, given the toxicity, candidates for IL-2 therapy should be carefully selected. The combination of IL-2 and IFN-alpha does not appear to provide additional survival benefit. Efforts to further improve therapeutic outcome for patients with metastatic renal cell carcinoma should focus on understanding the underlying mechanisms of cytokine-induced tumor regression. PMID- 10685668 TI - The role of interleukin-2 in the management of stage IV melanoma: the EORTC melanoma cooperative group program. AB - PURPOSE: To review the current information available from the European Organization for Research and Treatment of Cancer (EORTC) programs on the use of interleukin (IL)-2 in stage IV melanoma patients. PATIENTS AND METHODS: A database from 631 patients treated within 27 trials with high-dose IL-2-based regimens was compiled to develop hypotheses and valid stratification factors for randomized trials. Subsequently, 126 patients were enrolled in a trial evaluating interferon alfa (IFN-alpha) and IL-2 with or without cisplatin, and 325 patients were enrolled in an ongoing EORTC trial (18951) to evaluate dacarbazine, cisplatin, and IFN-alpha, with or without IL-2. RESULTS: The database suggests long-term survival rates of 23% and a 5-year survival rate of 13% for patients receiving a combination of IFN-alpha and IL-2 with or without chemotherapy. The addition of chemotherapy improved response rate but not survival. The first randomized trial testing the role of cisplatin in a chemoimmunotherapy regimen for advanced melanoma revealed a palliative effect for cisplatin but no survival benefit. The current trial (EORTC 18951), which is testing the impact of IL-2 on survival, is still immature. In the translational research program, we have evidence that patients in continuous complete remission after IL-2-based treatment have evidence of residual disease by polymerase chain reaction assay and, at the same time, melanoma-reactive T cells are present in the peripheral blood. CONCLUSION: Mature results defining the role and, to some extent, the mechanism of IL-2 in advanced melanoma are emerging. PMID- 10685669 TI - Overview of interleukin-2 inhalation therapy. AB - PURPOSE: Locoregional administration of interleukin (IL)-2, which acts physiologically as a local hormone, is an effective therapeutic modality. Diverse preclinical and clinical models have described methods of administration that expose tumor tissues to continuously high levels of cytokines. Regional administration of IL-2 that does not raise intravascular IL-2 levels induces local and systemic immunomodulation and produces objective local tumor responses. Most importantly, regional therapy is much less toxic than systemic IL-2 therapy. PATIENTS AND METHODS: We review clinical experiences using inhaled IL-2 therapy for the treatment of pulmonary metastases in roughly 300 patients with a variety of primary tumors. This includes our own 10-year single-institution experience with inhaled IL-2 therapy in the treatment of 188 metastatic renal cell carcinoma patients with progressive pulmonary metastases. Patients in our clinic are treated with 18 to 36 million IU of recombinant IL-2, administered 90% by inhalation and 10% subcutaneously, until disease progression. A variety of doses and schedules of inhaled IL-2 have been investigated alone and in combination with systemic therapies. RESULTS: Inhalation of IL-2 has been reported to prevent progression of pulmonary and mediastinal metastases of metastatic renal cell carcinoma, breast and ovarian carcinoma, and melanoma. Inhaled IL-2 alone is well tolerated; a dose-dependent cough is the major adverse event. A significant dose dependent increase in lymphocytes and eosinophils has been observed in bronchoalveolar lavage in patients and animals. Dose and schedule can influence outcome. In a phase I trial using inhaled IL-2 alone in patients with a variety of primary malignancies, once-daily inhalation of IL-2 at doses up to 15 million IU/m2 was well tolerated but did not result in prolonged stabilization of pulmonary disease. In a multidose phase I trial, using 5-times-daily inhalation of natural IL-2, pulmonary lesions in three of 14 (21%) metastatic renal cell carcinoma patients responded, and a similar multicenter trial demonstrated a 29% response rate. Among 188 metastatic renal cell carcinoma patients treated with inhaled recombinant IL-2 at the Clinic Eppendorf, progression of pulmonary metastases was prevented in 68% of patients for a median duration of 7 months, and overall survival was significantly improved compared with expected survival (Elson's risk analysis; 17.2 vs 5.3 mo). All patients, including high-risk patients, appeared to benefit. Encouraging results have also been reported in patients with metastatic melanoma and gynecologic tumors when inhaled IL-2 was used as second-line therapy to treat pulmonary metastases. CONCLUSIONS: The efficacy of inhaled IL-2, alone or in combination with systemic immunotherapy, immunochemotherapy, or chemotherapy, has been documented in a variety of malignancies. All reports confirm low toxicity, thus providing important quality of-life benefits. Moreover, patients not eligible for systemic IL-2 therapy may be treated with inhalation therapy. PMID- 10685670 TI - Central auditory processing disorders and reduced motivation: three case studies. AB - The central auditory test results for three normal-hearing children who were initially diagnosed as having a central auditory processing disorder and learning disability are presented. They were referred to the authors for second-opinion consultations. Central auditory processing retesting was performed by the authors under the condition of no reinforcement and then the condition of reinforcement with the child's favorite food, hobby, or toy. For all three cases, the central auditory test scores improved markedly bilaterally under the condition of reinforcement as compared with the condition of no reinforcement. We hypothesize that the improvement was related to increased motivation associated with the reinforcement and that these children represented false-positive results on the central auditory test battery. Large-sample studies are needed to investigate the effect of reinforcement on test performance in children with reduced central auditory test scores. PMID- 10685671 TI - Reconsidering the limits of normal hearing. AB - A proposal is made that 15 dB HL, rather than 25 dB HL, be considered the upper limit of normal hearing sensitivity. This recommendation is based on an explanation of the change from an earlier philosophy and the fact that so many people with hearing levels that average less than 25 dB HL consider themselves to have hearing difficulty. Such reclassification of people with slight to mild hearing losses would dignify their clinical complaints and aid in counseling that would assist them with their hearing difficulties. PMID- 10685672 TI - Self-reported hearing handicap and audiometric measures in older adults. AB - As part of an epidemiologic study of hearing disorders in older adults, audiometric thresholds (250-20,000 Hz), word recognition performance (Northwestern University Auditory Test No. 6 word lists in quiet and in competing message), and Hearing Handicap Inventory for the Elderly-Screening (HHIE-S) scores were evaluated for 3178 adults ranging in age from 48 to 92 years. Overall, higher HHIE-S scores were more prevalent for older age groups and for greater degrees of hearing loss. After adjusting for the degree of hearing loss, the probability of reporting a hearing disability (handicap) decreased with age. PMID- 10685673 TI - Self-perceived balance disability/handicap in the presence of bilateral peripheral vestibular system impairment. AB - The purpose of this report was to characterize the self-perceived balance disability/handicap of patients with bilateral reductions and bilateral complete losses of peripheral vestibular system function. Data from 72 patients whose electronystagmography and rotational examinations suggested normal, unilateral, or bilateral reductions in peripheral vestibular system function were used in the first investigation. Patients also completed a Dizziness Handicap Inventory (DHI). Results demonstrated significant group differences for DHI total and physical subscale scores. There were significant differences between normal and bilateral weakness groups for the total DHI score and between normal and unilateral and normal and bilateral weakness groups for the physical subscale score. In a second investigation, an item analysis of the DHI is presented for five patients with bilateral complete losses of peripheral vestibular system function. Results show that, predictably, these patients have difficulty engaging in activities requiring an intact vestibulocular reflex (e.g., physical activities such as sports, household chores). PMID- 10685674 TI - Acquiring counseling skills in mid-career: outcomes of a distance education course for practicing audiologists. AB - Although considered an essential component of audiology service delivery, counseling skills often are not adequately addressed in graduate training programs. One of the many goals of the audiology doctorate is to address this deficit by providing formal course work in counseling for both traditional graduate students and practitioners. This study describes the outcomes of a counseling class taught via distance education to mid-career audiologists, who initially were found to provide informational or technical responses to affective patient comments (a so-called "communication mismatch"). In spite of inherent limitations in electronic instruction, students demonstrated learning outcomes comparable to those obtained from conventional classroom instruction, including an improvement in their ability to respond to the affective nature of patient comments. PMID- 10685675 TI - Interlist equivalency of the Northwestern University Auditory Test No. 6 in quiet and noise with adult hearing-impaired individuals. AB - The purpose of the study was to determine the influence of sensorineural hearing loss and broadband noise on the interlist equivalency of the Northwestern University Auditory Test No. 6 (NU-6). There were two groups of participants: the first group consisted of 14 adults with mild-to-moderate hearing loss (mean age = 56 years; SD = 4.83); the second group consisted of 11 age-matched, normal hearing individuals (mean age = 55 years; SD = 4.69). Each group heard the four lists of the NU-6 in quiet and in broadband noise at four signal-to-noise ratios (-10 dB, -5 dB, 0 dB, and +5 dB). The NU-6 stimuli were presented at 35-dB sensation level relative to each listener's speech reception threshold. Results indicated that, for both groups, there was a significant main effect for NU-6 list. Post hoc single degree of freedom contrasts revealed that this main effect was due to significant differences between some of the lists when presented in background noise. There were no differences between the lists in quiet. Because of the findings of differences between some of the lists in noise, the authors suggested that clinicians or researchers use caution when comparing scores obtained from two different NU-6 lists over time. That is, if scores from two lists are different, it is important for the clinician to determine whether this disparity is due to a change in word recognition ability or simply due to a difference between the lists. PMID- 10685676 TI - Developmental changes in aural acoustic admittance measurements. AB - Tympanometry is a clinical measurement routinely included in the assessment of middle ear function. Despite its widespread use, however, fundamental questions remain regarding the need for age-dependent normative data. This study examines normal developmental changes associated with four tympanometric measurements: (1) ear canal volume, (2) peak compensated acoustic admittance, (3) tympanometric width, and (4) tympanometric peak pressure. Of 221 infants and children, aged 6 months to 5 years, enrolled in this study, 99 met the criteria for normal middle ear function as determined via pneumatic tympanoscopy by an experienced pediatric otolaryngologist, and data analysis was confined to those 99 volunteers. Analysis of variance revealed statistically significant main effects showing increases in ear canal volume and peak compensated acoustic admittance and decreases in tympanometric width as age increased. Statistically significant differences were not achieved for tympanometric peak pressure. Although statistically significant differences were found, the differences were small and of questionable clinical significance. PMID- 10685677 TI - Twenty-stimulus train for rapid acquisition of auditory brainstem responses in humans. AB - This study addressed the clinical need to obtain frequency-specific auditory brainstem responses (ABRs) more rapidly than is currently possible. ABRs were obtained from 20 subjects using two different methods: a conventional method with tone bursts presented singly and a multiple-stimulus method using a train of 20 tone bursts. For both methods, tone bursts were presented at frequencies 1, 2, 4, and 8 kHz, shaped with a Blackman-Harris window and having intensity levels up to 105 dB peak equivalent sound pressure level (peSPL). The single tone bursts were presented at a 17.2/sec repetition rate. The 20 tone-burst train used the four frequencies at five intensities each and a repetition rate of 3.7/sec (separations between tone bursts of 9-12 msec, with 77 msec off-time between trains). Mean latencies and mean amplitudes for wave V were compared using t tests for each of 12 conditions (four frequencies, each at the three highest output levels). For latency, only one comparison was significantly different (2 kHz, 77 dB peSPL). Similarly, only one comparison was significant for amplitude (2 kHz, 97 dB peSPL). There was, however, a trend for the tone bursts presented in trains to have longer latencies and reduced amplitudes compared to the respective responses for the single tone-burst condition. These results indicate the presence of some response adaptation when tone bursts are presented in a train. The use of a properly designed stimulus train can result in a significant time savings for obtaining frequency-specific ABRs as compared with single tone burst presentations. PMID- 10685678 TI - Identification of matrix metalloproteinases in canine neoplastic tissue. AB - OBJECTIVE: To identify matrix metalloproteinases (MMP) 2 and 9 in canine tumor tissue and to compare the amount of activity to that in unaffected stromal tissue. ANIMALS: 30 dogs with spontaneously developing, high-grade osteosarcoma. PROCEDURE: Tumor and nearby stromal tissue (muscle) were obtained at the time of surgery. Specimens were homogenized, and supernatants were assayed, using gelatin zymography. Human derived standards were run concurrently. Densitometry was done to obtain a semiquantitative arbitrary unit value for each specimen. The amount of activity in tumor tissue was compared with the amount in stromal tissue. RESULTS: Gelatinolytic bands were observed from the analysis of all tumor tissues and in most stromal tissues. These bands migrated in the same molecular weight area as the human MMP 2 and 9 standards. Gelatinolytic activity could be quenched by the addition of 50 mM EDTA and 1 microg of synthetic tissue inhibitor of metalloproteinase (TIMP) 2 per 100 ml. There was significantly more gelatinolytic activity in tumor tissue than in stromal tissue. CONCLUSIONS AND CLINICAL RELEVANCE: MMP 2 and 9 are detectable in canine neoplastic tissue. Matrix metalloproteinases activity in tumor tissue is higher than in unaffected stromal tissue, indicating that canine MMP may be involved in the pathogenesis of tumor growth and metastasis. PMID- 10685679 TI - Inheritance of von Willebrand's disease in a colony of Doberman Pinschers. AB - OBJECTIVE: To determine the mode of inheritance of von Willebrand's disease (vWD) and perform linkage analysis between vWD and coat color or narcolepsy in a colony of Doberman Pinschers. ANIMALS: 159 Doberman Pinschers. PROCEDURE: von Willebrand factor antigen (vWF:Ag) concentration was measured by use of ELISA, and results were used to classify dogs as having low (< 20%), intermediate (20 to 65%), or high (> 65%) vWF:Ag concentration, compared with results of analysis of standard pooled plasma. Buccal bleeding time was measured, and mode of inheritance of vWD was assessed by pedigree analysis. RESULTS: von Willebrand's disease was transmitted as a single autosomal gene defect. Results suggested that 27.04% of dogs were homozygous for vWD, 62.26% were heterozygous, and 10.69% did not have the defect. Most homozygous and some heterozygous dogs had prolonged bleeding times. Dogs with diluted coat colors (blue and fawn) were significantly overrepresented in the homozygous group, compared with black and red dogs, but a significant link between vWD and coat color was not detected. CONCLUSIONS AND CLINICAL RELEVANCE: von Willebrand's disease is transmitted as an autosomal dominant trait with variable penetrance; most dogs in this colony (89.3%) were carriers of vWD. Homozygosity for vWD is not likely to be lethal. Some heterozygous dogs have prolonged bleeding times. An association between diluted coat colors and vWD may exist. PMID- 10685680 TI - Effects of general anesthesia and surgery on renal function in healthy dogs. AB - OBJECTIVES: To evaluate renal function in healthy dogs undergoing general anesthesia and ovariohysterectomy without concurrent IV administration of fluids. ANIMALS: 35 healthy client-owned dogs. PROCEDURE: Dogs were medicated with promazine hydrochloride (0.05 mg/kg of body weight, SC) approximately 45 minutes before induction of anesthesia with thiopental sodium (10 to 15 mg/kg, IV). Anesthesia was maintained with 2% halothane in oxygen. Ovariohysterectomies were performed by senior veterinary students under the direct supervision of a veterinary surgeon. Renal function was assessed (serum urea and creatinine concentrations, fractional clearance of sodium, urine alkaline phosphatase [ALP] and gamma-glutamyltransferase [GGT] activities, urine specific gravity, and enumeration of renal tubular epithelial cells in urine sediment) prior to and 24 and 48 hours after surgery. RESULTS: Duration of general anesthesia ranged from 80 to 310 minutes. Urine specific gravity and ALP activity and serum urea and creatinine concentrations did not change over time. Fractional clearance of sodium decreased 24 and 48 hours after surgery, whereas urine GGT activity and the ratio of urine GGT activity to urine creatinine concentration increased 24 hours after surgery, compared with presurgery values. Renal tubular epithelial cells increased in number in urine sediment from 11 of 35 (31.4%) dogs and 5 of 35 (14.3%) dogs 24 and 48 hours after surgery, respectively. However, this increase was not clinically relevant. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous administration of fluids to healthy dogs undergoing general anesthesia and elective surgery may not be necessary for maintenance of renal homeostasis. PMID- 10685681 TI - Transformation and transposition of the genome of Mycobacterium marinum. AB - OBJECTIVE: To develop and evaluate protocols for genetic manipulations (transformation and transposition) of the fish pathogen, Mycobacterium marinum. SAMPLE POPULATION: Isolates of M. marinum obtained from fish and humans. PROCEDURE: Electrocompetent cells were prepared from isolates of M. marinum grown to various growth phases at several temperatures and with or without the addition of ethionamide or cycloheximide. Mycobacterial cells were transformed by electroporation with a replicative Escherichia coli-mycobacteria shuttle vector (pYUB18) as well as suicide vectors (pYUB285 and pUS252) that carried transposable elements (IS1096 and IS6110, respectively). Mutants from both isolates of M. marinum were recovered on 7H10 agar plates supplemented with kanamycin. Transformation and transposition efficiencies for various protocols were compared. Southern hybridization analysis was performed on mycobacterial mutants to confirm transposition events. RESULTS: Competent cells prepared at room temperature (23-25 C) from organisms in late-exponential growth phase yielded higher transposition efficiency, compared with cells prepared at 4 C or from organisms in early- or mid-exponential growth phase. Naturally developing kanamycin-resistant colonies of M. marinum were not detected. Only the IS1096 derived transposition was able to efficiently mutate M. marinum. Southern hybridization of M. marinum mutants revealed random integration of IS 1096 into the M. marinum genome. CONCLUSIONS: Transposition and transformation efficiencies were comparable, suggesting that the limiting factor in transposition is the transformation step. Most of the experiments resulted in transposition of IS1096; however, better approaches are needed to improve transposition efficiency. PMID- 10685682 TI - Development of an in vitro fluorometric assay to study adherence of Pasteurella haemolytica to bovine cells. AB - OBJECTIVE: To develop an in vitro fluorometric assay to assess Pasteurella haemolytica adherence to bovine respiratory and epithelial cells and compare adherence of single strains of P. haemolytica serovars A1 and A2 (PhA1 and PhA2, respectively). SAMPLE POPULATION: Monolayers of bovine turbinate and Madin Darby bovine kidney (MDBK) cells. PROCEDURE: To determine optimal inoculum concentration and incubation time, various concentrations of P. haemolytica were labeled with fluorescein isothiocyanate and incubated with monolayers of bovine cells for various times. Bovine cells were washed to remove nonadherent bacteria, and percentage of bacteria adhered (percentage of adherence) was estimated fluorometrically. Percentage of adherence of PhA1 was compared with that of PhA2. RESULTS: The optimal inoculum concentration that resulted in measurable fluorescence of adherent bacteria was 1 x 10(8) colony-forming units/ml, and the optimal incubation time was 45 minutes. Percentage of adherence of PhA1 to MDBK and turbinate cells was significantly greater than that determined for PhA2. CONCLUSIONS: The in vitro fluorometric assay is a time-efficient, inexpensive, and labor-saving method for evaluation of P. haemolytica adherence to bovine cells. The concentration of bacteria used to inoculate bovine cells in this assay is similar to that typically used in other types of in vitro adherence assays. The predominance of PhA1 over PhA2 during the early stages of bovine respiratory disease may be attributable to the ability of PhA1 to adhere more avidly to nasopharyngeal tissue. PMID- 10685683 TI - Comparison of sensitivity of sodium currents to tetrodotoxin in equine muscle specimens with that in murine and human muscle specimens. AB - OBJECTIVE: To determine sensitivity of equine skeletal muscle to tetrodotoxin and compare that with sensitivity of murine and human skeletal muscles. SAMPLE POPULATION: Semimembranosus, vastus lateralis, triceps brachii, and masseter muscle specimens from 22 euthanatized horses, vastus lateralis muscle biopsy specimens from 25 clinically normal humans, and diaphragmatic muscle specimens from 6 mice. PROCEDURE: Electrically elicited twitch responses were measured in muscle specimens incubated in medium alone and with tetrodotoxin (100 nM, 400 nM, 1.6 microM for equine specimens and 100 nM, 200 nM, 400 nM, 800 nM, 1.6 microM for murine and human specimens). Percentages of tetrodotoxin-sensitive and resistant sodium channels were determined and compared among muscles and species. RESULTS: 2 sodium channels with different sensitivities to tetrodotoxin were identified in equine muscle. One was blocked with 100 nM tetrodotoxin and the other was unaffected by tetrodotoxin at concentrations up to 1.6 microM. The only difference detected among the 4 equine muscles was that masseter muscle specimens had a higher percentage of tetrodotoxin-sensitive channels than triceps brachii muscle specimens. Tetrodotoxin-resistant sodium channels constituted 31 to 66% of the equine muscle twitch response, which was greater than that determined for normal human and murine muscle (< 5%). CONCLUSION AND CLINICAL RELEVANCE: Equine skeletal muscle contains a high percentage of tetrodotoxin-resistant sodium channels. The 4 equine muscles evaluated were more similar to each other than to murine and human muscles. Shifts in expression of sodium channel subtypes may play a role in the manifestation of certain myopathies. PMID- 10685684 TI - Histologic and immunohistochemical characterization of lens capsular plaques in dogs with cataracts. AB - OBJECTIVE: To determine histologic and immunohistochemical characteristics of the multifocal adherent plaques that commonly develop on the internal surfaces of the anterior and posterior lens capsules in dogs with cataracts. SAMPLE POPULATION: 31 anterior and 4 posterior capsular specimens collected during lens extraction surgery in dogs with cataracts. PROCEDURE: Specimens were evaluated, using light and transmission electron microscopy. Immunohistochemical techniques were used to localize cytokeratin, vimentin, alpha-smooth muscle-specific actin, fibronectin, tenascin, and transforming growth factor-beta (TGF-beta) within plaques. RESULTS: Histologically, plaques comprised elongated spindle-shaped cells that formed a placoid mass. Cells were embedded in an extracellular matrix containing collagen fibrils, often with duplicated or split basement membranes. Immunohistochemically, normal lens epithelial cells and cells within plaques stained for vimentin. Most cells and some areas of the extracellular matrix within plaques stained for TGF-beta and alpha-smooth muscle-specific actin. Fibronectin and tenascin were also detected in the extracellular matrix. CONCLUSIONS AND CLINICAL RELEVANCE: Canine lens capsular plaques are histologically and immunohistochemically similar to posterior capsule opacification and subcapsular cataracts in humans, which suggests that the canine condition, like the human conditions, is associated with fibrous metaplasia of lens epithelial cells. Transforming growth factor-beta may play a role in the genesis of capsular plaques. Because severity of plaques was correlated with stage of cataract development, earlier surgical removal of cataracts may be useful to avoid complications associated with plaque formation. PMID- 10685685 TI - Determination of the lactate breakpoint during incremental exercise in horses adapted to dietary corn oil. AB - OBJECTIVE: To determine lactate breakpoint of horses and test for effects of training and dietary supplementation with corn oil on that breakpoint. ANIMALS: 7 healthy Arabian horses. PROCEDURES: Horses received a control diet (n = 4) or a diet supplemented with 10% corn oil (4). A training program, which comprised two 5-week conditioning periods with 1 week of rest, was initiated. Submaximal incremental exercise tests (IET) were conducted before the first and after both conditioning periods. Blood samples for determination of blood lactate and plasma glucose concentrations were collected 1 minute before IET and during the 15 seconds immediately preceding each speed change. Data collected were fit to one- and two-slope broken-line models and an exponential model. RESULTS: Good fits were obtained by application of the broken-line models (adjusted R2 > 0.92) to blood lactate concentration versus speed curves. Lactate breakpoints increased 41% after training. After training, slope 2 and peak blood lactate concentrations were greater in the corn oil group, compared with controls. Mean blood lactate concentration at the breakpoint was not affected by training or diet. Plasma glucose concentration versus speed curves also fit the broken-line models, and glucose breakpoints preceded lactate breakpoints by approximately 1 m/s in the second and third IET. CONCLUSIONS AND CLINICAL RELEVANCE: Lactate breakpoints can be determined for horses, using blood lactate concentration versus speed curves generated during submaximal IET and may be useful for assessing fitness and monitoring training programs in equine athletes. PMID- 10685686 TI - Hepatic and pulmonary enzyme activities in horses. AB - OBJECTIVE: To determine hepatic and pulmonary phase-I and phase-II enzyme activities in horses. SAMPLE POPULATION: Pulmonary and hepatic tissues from 22 horses that were 4 months to 32 years old. PROCEDURE: Pulmonary and hepatic tissues from horses were used to prepare cytosolic (glutathione S-transferase and soluble epoxide hydrolase) and microsomal (cytochrome P450 monooxygenases) enzymes. Rates of microsomal metabolism of ethoxyresorufin, pentoxyresorufin, and naphthalene were determined by high-performance liquid chromatography. Activities of glutathione S-transferase and soluble epoxide hydrolase were determined spectrophotometrically. Cytochrome P450 content was determined by carbon monoxide bound-difference spectrum of dithionite-reduced microsomes. Activity was expressed relative to total protein concentration. RESULTS: Microsomal protein and cytochromeP450 contents were detectable in all horses and did not vary with age. Hepatic ethoxyresorufin metabolism was detected in all horses; by comparison, pulmonary metabolism of ethoxyresorufin and hepatic and pulmonary metabolism of pentoxyresorufin were detected at lower rates. Rate of hepatic naphthalene metabolism remained constant with increasing age, whereas rate of pulmonary naphthalene metabolism was significantly lower in weanlings (ie, horses 4 to 6 months old), compared with adult horses. Hepatic glutathione S-transferase activity (cytosol) increased with age; however, these changes were not significant. Pulmonary glutathione S-transferase activity (cytosol) was significantly lower in weanlings than adult horses. Hepatic and pulmonary soluble epoxide hydrolase did not vary with age of horses. CONCLUSIONS AND CLINICAL RELEVANCE: Activity of cytochrome P450 isoforms that metabolize naphthalene and glutathione S-transferases in lungs are significantly lower in weanlings than adult horses, which suggests reduced ability of young horses to metabolize xenobiotics by this organ. PMID- 10685687 TI - Fungal flora on cutaneous and mucosal surfaces of cats infected with feline immunodeficiency virus or feline leukemia virus. AB - OBJECTIVE: To compare cutaneous and mucosal mycoflora in cats infected with FIV or FeLV with that in noninfected cats. ANIMALS: 85 client-owned cats; 24 seropositive for FIV, 10 seropositive for FeLV, 1 seropositive for both viruses, and 50 seronegative for both viruses. PROCEDURE: Cutaneous specimens were obtained from the coat and external acoustic meatus (ear canal) and mucosal specimens from the oropharynx and rectum. Fungi were isolated from specimens, using Sabouraud dextrose agar incubated at 27 or 37 C for cutaneous and mucosal specimens, respectively. RESULTS: Fungal colonies were cultured from at least 1 specimen from 83 of 85 (97.6%) cats. The most common fungal isolates were Aspergillus spp (cultured from 59.3% of all specimens), Penicillium spp (50.0%), Cladosporium spp (44.2%), Scopulariopsis spp (41.8%), and lipophilic yeasts of the genus Malassezia (31.4%). A greater diversity of fungal genera was isolated from retrovirus-infected cats, and Malassezia spp were more commonly recovered from these cats, compared with noninfected cats. Candida albicans, Cryptococcus neoformans, and dermatophytes (eg, Microsporum canis) were rarely isolated from any cat. Significant differences in frequency of isolation of C. neoformans and dermatophytes were not found between infected and noninfected cats. CONCLUSIONS AND CLINICAL RELEVANCE: Cats infected with FIV or FeLV may have a greater diversity of cutaneous and mucosal mycoflora than noninfected cats. However, infected cats may be no more likely than noninfected cats to expose humans to zoonotic fungi such as C. albicans, C. neoformans, and M. canis. PMID- 10685688 TI - Comparison of the phenotypes of Streptococcus zooepidemicus isolated from tonsils of healthy horses and specimens obtained from foals and donkeys with pneumonia. AB - OBJECTIVE: To determine whether streptococcal pneumonia is caused by strains of Streptococcus zooepidemicus similar to those obtained from the tonsils of healthy horses. SAMPLE POPULATION: 5 tonsils from healthy horses, 8 tracheal washes and 6 lung specimens from foals with pneumonia, and 5 nasopharyngeal swab specimens from donkeys with acute bronchopneumonia. PROCEDURE: Variable M-like protectively immunogenic SzP proteins of 5 isolates of S. zooepidemicus from each tonsil and clinical specimen were compared, using immunoblots. The SzP gene of 13 isolates representative of various SzP immunoblot phenotypes from 1 healthy horse and 9 horses and donkeys with pneumonia were sequenced and compared. Cell-associated hyaluronic acid concentration and resistance to phagocytosis of some isolates were measured. RESULTS: Tonsils of each healthy horse were colonized by several SzP phenotypes similar to those of foals or donkeys with pneumonia. In contrast, multiple isolates from animals with pneumonia had the same SzP phenotype, indicating infection by a single strain or clone. Analysis of the SzP sequence confirmed that differences in immunoblot phenotype were associated with sequence differences and that several SzP genotypes were in healthy horses and animals with pneumonia. Isolates with high concentrations of cell-associated hyaluronic acid were more resistant to phagocytosis. CONCLUSIONS AND CLINICAL RELEVANCE: An SzP-specific immunoblot is a useful, sensitive measure of diversity among strains of S. zooepidemicus. Single strains with SzP phenotypes similar to those found in tonsils of healthy horses cause pneumonia. Because of the diversity of SzP phenotype and genotype among isolates from animals with pneumonia, SzP phenotype is not an important determinant of invasiveness or epizootic capabilities. PMID- 10685689 TI - Correlation of clinical score, intrapleural pressure, cytologic findings of bronchoalveolar fluid, and histopathologic lesions of pulmonary tissue in horses with summer pasture-associated obstructive pulmonary disease. AB - OBJECTIVE: To correlate clinical score, intrapleural pressure, cytologic findings of bronchoalveolar lavage fluid (BALF), and histologic lesions of pulmonary tissue in horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD). ANIMALS: 8 adult horses affected with SPAOPD and 6 adult horses without evidence of respiratory tract disease. PROCEDURE: Clinical score, change in intrapleural pressure (deltaPpl) during tidal breathing, results of cytologic examination and bacteriologic culture of BALF, and results of histologic examination of pulmonary parenchyma were evaluated. RESULTS: Clinical scores for SPAOPD-affected horses (median, 5.75; range, 4.0 to 7.5) were significantly greater, compared with clinically normal horses (median, 2.0; range, 2.0 to 3.0). Cytologic examination of BALF from SPAOPD-affected horses revealed predominantly nondegenerate neutrophils. Histologic lesions were identified throughout pulmonary tissue and included severe accumulation of mucus and neutrophils within the small airways, metaplasia of bronchiolar goblet cells, and mild peribronchial infiltrate. Histologic examination of specimens collected via percutaneous biopsy was predictive of disease and corresponded to findings at postmortem examination. Clinical score and deltaPpl were highly correlated with mucus accumulation in the airways of affected horses. Peribronchial inflammatory infiltrate correlated with percentage of neutrophils in BALF of affected horses. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical scoring and deltaPpl provided valid estimates of disease severity. Findings from cytologic examination of BALF of SPAOPD-affected horses varied, although, in most instances, it was diagnostically useful. Severe mucus accumulation in the airways was the most remarkable histopathologic finding in SPAOPD-affected horses. Examination of biopsy specimens collected from pulmonary parenchyma was consistently useful in diagnosing SPAOPD. PMID- 10685690 TI - Ultrastructural changes in follicles of small-intestinal aggregated lymphoid nodules in early and advanced phases of experimentally induced mucosal diseases in calves. AB - OBJECTIVE: To investigate ultrastructural changes in follicles of small intestinal aggregated lymphoid nodules (Peyer's patches) of calves with early and advanced phases of experimentally induced mucosal disease (MD). ANIMALS: Twenty 2.5- to 7-month-old Holstein-Friesian calves (11 females, 9 males). PROCEDURE: MD was induced in 13 of 18 calves that were persistently viremic with bovine viral diarrhea virus (BVDV). Eight of the 13 calves were euthanatized before the onset of clinical signs of MD, and 5 were euthanatized after becoming moribund with MD. Five persistently viremic calves and 2 calves without BVDV served as controls. Specimens of small-intestinal aggregated lymphoid nodules were prepared for transmission electron microscopy. RESULTS: The ultrastructure of follicles of small-intestinal aggregated lymphoid nodules from healthy calves was consistent with that in sheep. In the early phase of MD, changes were characterized by numerous apoptotic lymphocytes and macrophages with apoptotic bodies. In more advanced lesions, affected lymphoid follicles consisted of macrophages and variable numbers of follicular dendritic cells (FDC), whereas others did not contain FDC. In moribund calves, small follicles consisting predominantly of FDC and follicles with central cavities surrounded by macrophages, and few neutrophils were observed. CONCLUSIONS AND CLINICAL RELEVANCE: The ultrastructural changes in lymphoid follicles of small-intestinal aggregated lymphoid nodules indicate apoptosis of lymphocytes as an initial event. The development of small follicles consisting predominantly of FDC or the complete loss of follicular architecture in advanced phases of MD is determined by the intensity of apoptosis of lymphocytes, the capacity of the macrophages for uptake, and the reorganization of a stromal network. PMID- 10685691 TI - Doppler ultrasonographic features of thoracic limb arteries in clinically normal horses. AB - OBJECTIVES: To determine blood flow velocities and indices from spectral waveforms obtained by use of Doppler ultrasonography of thoracic limb arteries of horses and to assess interobserver and patient variability associated with the technique. ANIMALS: 9 clinically normal adult horses. PROCEDURE: Left thoracic limb arteries of 8 nonsedated horses were examined at 5 sites by use of pulsed wave Doppler ultrasonography to determine a range of values for peak systolic, end diastolic, and mean velocities and resistive and pulsatility indices. Interobserver and patient variabilities were determined by 2 operators repeating similar measurements on 1 horse 8 times at weekly intervals. RESULTS: A range of values for each variable measured at the 5 selected sites was obtained. For each variable, strong positive correlations (R > or = 0.7) were detected for > 70% of the site-to-site comparisons made (excluding the coronary band). Among horses, resistive index varied least, whereas over time, mean velocity varied least. Waveform characteristics were consistent with resistive (n = 5) or nonresistive (4) patterns. In the single-horse experiment, waveform characteristics were consistent throughout the 8 weeks, and operator effects were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Doppler ultrasonography of no one site resulted in more reliable measurements of blood flow characteristics in thoracic limb arteries of horses. Mean velocity and resistive index were the least variable measurements made. Pulsed-wave Doppler ultrasonography may be a useful technique for evaluating diseases that alter normal thoracic limb arterial blood flow in horses. PMID- 10685692 TI - Kinematics and ground reaction forces in horses with superficial digital flexor tendinitis. AB - OBJECTIVE: To measure and correlate kinematic and ground reaction force (GRF) data in horses with superficial digital flexor tendinitis. ANIMALS: 6 sound horses. PROCEDURE: Horses were evaluated before (sound evaluation) and after (lame evaluation) induction of superficial digital flexor tendinitis in 1 forelimb (randomized) by injection of collagenase. As each horse trotted, kinematic data were collected by use of an optoelectronic system, and GRF data were measured by use of a force plate. Three-dimensional kinematic and GRF data were projected onto a 2-dimensional sagittal plane. RESULTS: Lame limbs had significantly lower peak vertical GRF, less flexion of the distal interphalangeal joint, and less extension of the metacarpophalangeal joint, compared with compensating limbs. Carpal joint kinematics did not change. Compensating limbs had a more protracted orientation throughout the stance phase and higher braking longitudinal force and impulse; however, total range of rotation from ground contact to lift off did not change. Transfer of body weight from lame to compensating limbs was smooth, without elevation of the body mass into a suspension phase. Propulsive components of longitudinal GRF did not differ between limbs. CONCLUSIONS AND CLINICAL RELEVANCE: In horses with experimentally induced superficial digital flexor tendinitis, changes in vertical GRF were reflected in angular excursions of the distal interphalangeal and metacarpophalangeal joints, whereas changes in longitudinal GRF were associated with alterations in the protraction-retraction angle of the entire limb. PMID- 10685693 TI - Net joint moments and joint powers in horses with superficial digital flexor tendinitis. AB - OBJECTIVE: To determine whether analysis of net joint moments and joint powers is a suitable technique for evaluation of mechanics and energetics of lameness in horses and to measure effects of superficial digital flexor tendinitis. ANIMALS: 6 sound horses. PROCEDURE: Horses were evaluated before (sound evaluation) and after (lame evaluation) induction of superficial digital flexor tendinitis in 1 forelimb by injection of collagenase. Recordings were made with an optoelectronic system and a force plate as horses trotted. Net joint moments and joint powers in the sagittal plane at each joint in the forelimbs during the stance phase were determined. Peak values were determined, and mechanical energy absorbed and generated at each joint was calculated. Comparisons were made between contralateral limbs during sound and lame evaluations. RESULTS: Lame limbs had significant reductions in peak values for net joint moments on the palmar aspect of metacarpophalangeal (fetlock), carpal, and humeroulnar joints. Total energy absorbed was significantly lower at every joint in lame limbs, compared with compensating limbs. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with superficial digital flexor tendinitis had significant differences between lame and compensating limbs for net joint moments and joint powers at all joints, indicating that the gait of horses with superficial digital flexor tendinitis is energetically inefficient. Assessment of net joint moments and joint powers is a useful tool in evaluating equine lameness. PMID- 10685694 TI - Structure-related echoes in ultrasonographic images of equine superficial digital flexor tendons. AB - OBJECTIVE: To develop a method to discriminate between structure-related echoes and echoes resulting from interference, as observed in transverse ultrasonographic images of equine superficial digital flexor (SDF) tendons. SAMPLE POPULATION: 2 normal (injury-free) SDF tendons obtained from a 3-year-old Thoroughbred and a 9-year-old Dutch Warmblood horse. PROCEDURE: Tendons were mounted in a custom-made device that permitted exact transverse and perpendicular sequential scanning with precise steps of 0.5 mm along the long axis of the tendon. Photographs of transverse tendon sections at the exact scanning locations were obtained. Propagation, reflection, and refraction artifacts were quantified, and an image rectification procedure was developed, allowing exact matching of each photograph with the corresponding ultrasonographic image. A correlation routine was developed that departed from this transverse ultrasonographic image (position 0); this routine added information from images collected at precise distances of 0.5 and 1 mm on both sides of the actual scan location (positions 2, -1, +1, +2). RESULTS: By use of the correlation routine, echoes that remained steady over all 5 images were enhanced and resolved, and constantly changing echoes were multiplicatively reduced and faded. This correlated image could be projected over the rectified photograph, and the resolved echoes matched perfectly with the endotendon septa surrounding fibers and fasciculi. CONCLUSIONS AND CLINICAL RELEVANCE: The correlation routine permits exclusive resolution of structure-related echoes, as echoes resulting from interference are faded. The technique described can produce images that depict only the essential structure related information. In this way, the clinical assessment of tendon integrity is greatly facilitated. PMID- 10685695 TI - Ultrasonographic tissue characterization of equine superficial digital flexor tendons by means of gray level statistics. AB - OBJECTIVE: To correlate quantitative analysis of ultrasonographic images of normal (injury-free) equine superficial digital flexor (SDF) tendons and equine SFD tendons that have pathologic changes with corresponding histologic sections. SAMPLE POPULATION: 4 SDF tendons, 2 of which had various stages of tissue integrity. The 2 ipsilateral tendons were used as points of reference. PROCEDURE: Tendons were mounted in a custom-made device that permitted sequential scanning, transversely and perpendicular to the tendon long axis. At precise steps of 0.5 mm, transverse ultrasonographic images were collected. Subsequently, tendons were fixed and prepared for histologic examination. The following 8 tissue types were discerned: normal young, normal old, necrotic, early granulation, late granulation, early fibrotic, late fibrotic, and scar tissues. In areas of interest, the corresponding ultrasonographic images were selected for gray level statistical analysis. RESULTS: Compared with other tissue types, early-stage granulation tissue was characterized by substantially lower mean gray level and a clearly different histogram. Necrotic tissue had a higher mean gray level, with a virtually normal histogram. In late granulation and early fibrotic tissues, the mean gray level and the histogram could not be discerned from those of normal tendon tissue. The same applied to late fibrotic and scar tissues; mean gray levels were fractionally lower than those of normal tendon tissue with a completely normal histogram. CONCLUSIONS: Although quantification of the transverse ultrasonographic image by use of first-order gray level statistics may be helpful, the method is not sufficiently sensitive to accurately and unequivocally determine the type of tendon tissue. Quantitative analysis should incorporate transverse and longitudinal information. PMID- 10685696 TI - Indications for emergent MRI of the central nervous system. PMID- 10685698 TI - Implant recommendations. PMID- 10685697 TI - Innovation and service traditional at University of Michigan Medical School. PMID- 10685699 TI - Stem cell transplants found superior. PMID- 10685700 TI - From the Centers for Disease Control and Prevention. Recommended childhood immunization schedule--United States, 2000. PMID- 10685701 TI - From the Centers for Disease Control and Prevention. Hypothermia-related deaths- Alaska, October 1998-April 1999, and trends in the United States, 1979-1996. PMID- 10685702 TI - From the Centers for Disease Control and Prevention. Update: influenza activity- United States, 1999-2000 season. PMID- 10685703 TI - Brain injury in amateur soccer players. PMID- 10685704 TI - Brain injury in amateur soccer players. PMID- 10685705 TI - Access to antiretroviral therapy. PMID- 10685706 TI - Access to antiretroviral therapy. PMID- 10685707 TI - The herbal history of Digitalis: lessons for alternative medicine. PMID- 10685708 TI - The National Practitioner Data Bank and the quality of peer review. PMID- 10685709 TI - The National Practitioner Data Bank and the quality of peer review. PMID- 10685710 TI - The National Practitioner Data Bank and the quality of peer review. PMID- 10685711 TI - Does prenatal famine cause later antisocial behaviors? PMID- 10685712 TI - Does prenatal famine cause later antisocial behaviors? PMID- 10685713 TI - Cost-utility analysis of screening intervals for diabetic retinopathy in patients with type 2 diabetes mellitus. AB - CONTEXT: Annual eye screening for patients with diabetes mellitus is frequently proposed as a measure of quality of care. However, the benefit of annual vs less frequent screening intervals has not been well evaluated, especially for low-risk patients. OBJECTIVE: To examine the marginal cost-effectiveness of various screening intervals for eye disease in patients with type 2 diabetes, stratified by age and level of glycemic control. DESIGN: Markov cost-effectiveness model. SETTING AND PARTICIPANTS: Hypothetical patients based on the US population of diabetic patients older than 40 years from the Third National Health and Nutrition Examination Survey. MAIN OUTCOME MEASURES: Patient time spent blind, quality-adjusted life-years (QALYs), and costs of annual vs less frequent screening compared by age and level of hemoglobin A1c. RESULTS: Retinal screening in patients with type 2 diabetes is an effective intervention; however, the risk reduction varies dramatically by age and level of glycemic control. On average, a high-risk patient who is aged 45 years and has a hemoglobin A1c level of 11% gains 21 days of sight when screened annually as opposed to every third year, while a low-risk patient who is aged 65 years and has a hemoglobin A1c level of 7% gains an average of 3 days of sight. The marginal cost-effectiveness of screening annually vs every other year also varies; patients in the high-risk group cost an additional $40530 per QALY gained, while those in the low-risk group cost an additional $211570 per QALY gained. In the US population, retinal screening annually vs every other year for patients with type 2 diabetes costs $107510 per QALY gained, while screening every other year vs every third year costs $49760 per QALY gained. CONCLUSIONS: Annual retinal screening for all patients with type 2 diabetes without previously detected retinopathy may not be warranted on the basis of cost-effectiveness, and tailoring recommendations to individual circumstances may be preferable. Organizations evaluating quality of care should consider costs and benefits carefully before setting universal standards. PMID- 10685714 TI - The International Registry of Acute Aortic Dissection (IRAD): new insights into an old disease. AB - CONTEXT: Acute aortic dissection is a life-threatening medical emergency associated with high rates of morbidity and mortality. Data are limited regarding the effect of recent imaging and therapeutic advances on patient care and outcomes in this setting. OBJECTIVE: To assess the presentation, management, and outcomes of acute aortic dissection. DESIGN: Case series with patients enrolled between January 1996 and December 1998. Data were collected at presentation and by physician review of hospital records. SETTING: The International Registry of Acute Aortic Dissection, consisting of 12 international referral centers. PARTICIPANTS: A total of 464 patients (mean age, 63 years; 65.3% male), 62.3% of whom had type A dissection. MAIN OUTCOME MEASURES: Presenting history, physical findings, management, and mortality, as assessed by history and physician review of hospital records. RESULTS: While sudden onset of severe sharp pain was the single most common presenting complaint, the clinical presentation was diverse. Classic physical findings such as aortic regurgitation and pulse deficit were noted in only 31.6% and 15.1% of patients, respectively, and initial chest radiograph and electrocardiogram were frequently not helpful (no abnormalities were noted in 12.4% and 31.3% of patients, respectively). Computed tomography was the initial imaging modality used in 61.1%. Overall in-hospital mortality was 27.4%. Mortality of patients with type A dissection managed surgically was 26%; among those not receiving surgery (typically because of advanced age and comorbidity), mortality was 58%. Mortality of patients with type B dissection treated medically was 10.7%. Surgery was performed in 20% of patients with type B dissection; mortality in this group was 31.4%. CONCLUSIONS: Acute aortic dissection presents with a wide range of manifestations, and classic findings are often absent. A high clinical index of suspicion is necessary. Despite recent advances, in-hospital mortality rates remain high. Our data support the need for continued improvement in prevention, diagnosis, and management of acute aortic dissection. PMID- 10685715 TI - Extracorporeal life support: the University of Michigan experience. AB - The University of Michigan experience with extracorporeal life support (ECLS) in 1000 consecutive patients between 1980 and 1998 is the largest series at one institution in the world. Among this patient population, survival to hospital discharge in moribund patients with respiratory failure was 88% in 586 neonates, 70% in 132 children, and 56% in 146 adults. Survival in moribund patients with cardiac failure was 48% in 105 children and 33% in 31 adults. This article describes the University of Michigan's overall ECLS patient experience, the progression of ECLS from laboratory experiments to clinical application at the bedside, the expansion of the technology to other centers, and current ECLS technology and outcomes. Despite the challenges faced in clinical research in this field, our experience and that of others has shown that ECLS saves lives of patients with acute cardiac or pulmonary failure in a variety of clinical settings. PMID- 10685716 TI - Conflicts regarding decisions to limit treatment: a differential diagnosis. AB - Conflicts between physicians and families about end-of-life decisions create challenging and emotionally difficult situations. In this article, we propose a "differential diagnosis" of such conflicts, distinguishing and describing the characteristics of families, physicians, and organizations and society that contribute to the "etiology" of the situation, as well as strategies for "diagnosing" the dominant factors. As a medical model, the differential diagnosis can be a useful tool to help physicians understand and manage conflicts about end of-life care. PMID- 10685717 TI - The University of Michigan Medical School, 1850-2000: "an example worthy of imitation". AB - The 150th anniversary of the University of Michigan Medical School affords occasion for both celebration and reflection, not just in Ann Arbor but throughout the world, as we consider its contributions to medical education, research, and health care over the past century and a half. This article explores the medical school's origins as a frontier medical outpost and describes the vital reforms in medical education implemented in Ann Arbor long before the landmark Flexner Report on Medical Education of 1910. It also depicts how and why the medical school developed as it did and what features are distinctive or typical about the school during this period. PMID- 10685718 TI - ERISA litigation and physician autonomy. AB - The Employee Retirement Income Security Act (ERISA), enacted in 1974 to regulate pension and health benefit plans, is a complex statute that dominates the managed care environment. Physicians must understand ERISA's role in the relationship between themselves and managed care organizations (MCOs), including how it can influence clinical decision making and physician autonomy. This article describes ERISA's central provisions and how ERISA influences health care delivery in MCOs. We analyze ERISA litigation trends in 4 areas: professional liability, utilization management, state legislative initiatives, and compensation arrangements. This analysis demonstrates how courts have interpreted ERISA to limit physician autonomy and subordinate clinical decision making to MCOs' cost containment decisions. Physicians should support efforts to amend ERISA, thus allowing greater state regulatory oversight of MCOs and permitting courts to hold MCOs accountable for their role in medical decision making. PMID- 10685720 TI - JAMA Patient Page: eye health. PMID- 10685719 TI - Shaping a positive future for academic medicine at Michigan. PMID- 10685721 TI - Role of cardiovascular reactivity to mental stress in predicting future hypertension. AB - Hypertension (HT) has been known since times immemorial to be one of the major causes of morbidity and mortality. It contributes to atherosclerotic cardiovascular disease, increasing its risk 2-3 times and is also associated with dyslipidemia, insulin resistance, glucose intolerance and obesity (1). The age of onset of hypertension is now earlier than before, making it essential that early detection of people who could be future hypertensives is done. Therefore, cardiovascular reactivity to stress in predicting future hypertension becomes important. In this fast paced age most people are exposed to mental stress which is the most common and prevalent form of stress. Increase in blood pressure (BP) in response to emotional arousal is well known, but support for this hypothesis of reactivity in predicting future hypertension is limited. We are attempting here to put forth a review of the various endeavours done so far to support this hypothesis. PMID- 10685722 TI - Efficacy and safety of a fixed low-dose perindopril/indapamide combination in essential hypertension. A randomised controlled study. AB - This multicenter, double-blind, parallel-group study was designed to assess the efficacy and the safety of fixed low dose combination perindopril 2 mg/indapamide 0.625 mg (Per/Ind) versus atenolol 50 mg (Ate). After a 4-week placebo run-in, 446 hypertensive patients (mean age : 55.8 +/- 11.0 years) were randomised to receive Per/Ind or Ate for 12 weeks. The primary outcome measures were the changes in trough supine systolic and diastolic blood pressure (sSBP, sDBP) between baseline and the last observation. Equivalence was assessed in an intention-to-treat analysis using a two one-sided tests procedure. Per/Ind and Ate decreased sSBP by -20.5 mmHg and -20.1 mmHg, respectively; the 90% confidence interval [-2.3; 1.5] of the intertreatment difference (-0.4 mmHg) fell within the predefined equivalence interval [-8; +8 mmHg]. Similarly, the sDBP decreased by 15.1 mmHg (Per/Ind) and -16.2 mmHg (Ate) with an intertreatment difference of 1.1 mmHg whose 90% confidence interval [-0.1; 2.2 mmHg] fell within the predefined equivalence interval [-4; +4 mmHg]; thus antihypertensive efficacy of Per/Ind and Ate were equivalent (P <0.001). In patients older than 65, Per/Ind induces a statistically greater decrease in sSBP than Ate (P <0.05). Per/Ind was well tolerated. Further controlled studies are needed to confirm these results on a long-term period. PMID- 10685723 TI - Gender differences in growth of vascular smooth muscle cells isolated from hypertensive and normotensive rats. AB - Higher male sensitivity to atherosclerotic and hypertensive events was a reason to study sex differences in migration and proliferation of vascular smooth muscle cells (VSMC) isolated from male and female spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) controls. Outgrowth of cells from explants, doubling time, curves of cumulative labeling and the length of cell cycle were measured in aortic VSMC. Systolic and mean arterial pressures were higher in males than in females of the two strains. The migration of cells from male explants was significantly faster than those from female aortas in both strains. The doubling time was always shorter in male VSMC than in those from females and this was more apparent in the late exponential phase of growth. The thymidine incorporation into newly synthesized DNA, which was enhanced in SHR compared to WKY cells, was also higher in male cells compared to female ones. Cell cycle was always shorter in male than in female VSMC due to the shorter G1 phase. In contrast, shorter S phase caused shorter cell cycle in SHR compared to WKY VSMC. Consequently, the shortest cell cycle was found in VSMC from SHR males with the highest blood pressure. It can be concluded that gender and genotype are two independent factors participating in the control of migration and proliferation of VSMC. PMID- 10685724 TI - Effects of a neutral endoprotease enzyme inhibitor, thiorphan, on hemodynamics and renal excretory function in four models of experimental hypertension. AB - Thiorphan, a neutral endoprotease (NEP) enzyme inhibitor, has been shown to enhance the effects of atrial natriuretic peptide (ANP) in vivo. In this study, we examined the effects of an intravenous (iv) infusion of thiorphan on cardiovascular hemodynamics and excretion of urine volume (UV), sodium (U(Na)V) and potassium (UKV) in four different models of experimental hypertension, namely: 1) SHR, 2) two-kidney, one clip (2K1C),3) one-kidney, 1 clip (1K1C) and. 4) 70% reduced renal mass-salt (RRM-S) hypertensive rats. SHR has normal plasma renin activity, 2K1C is renin dependent, and 1K1C and RRM-S are low renin volume dependent models of hypertension. Rats were divided into experimental and control groups. Under inactin (120 mg/kg, body weight) anesthesia, rats were instrumented to record blood pressure and dP/dt (Millar catheter) and urine was collected through a suprapubic urinary bladder catheter. Experimental animals received an iv infusion of thiorphan, 0.5 mg/kg/min for 120 minutes. Control animals received vehicle only. In some animals, vascular smooth muscle cell membrane potentials (Em) was measured in vivo. In another series of experiments, using the identical protocol, cardiac output was recorded. The thiorphan infusion produced a similar progressive decrease in blood pressure in all models of hypertension. Cardiac output did not change relative to vehicle infused control animals. Thus pressure decreased because of a decrease in total peripheral resistance. The contractility index (dP/dt/P, where P = left ventricular pressure) did not change but vascular smooth muscle cells in tail arteries hyperpolarized in all four models. In spite of a significant decrease in blood pressure, thiorphan infusion either increased or produced no change in urinary volume (UV) and sodium (U(Na)V) excretion. These data show that thiorphan, an NEP inhibitor, decreases the blood pressure of hypertensive rats due to a decrease in total peripheral resistance, perhaps by hyperpolarizing vascular smooth muscle cells. These effects are independent of the mechanism of the hypertension. Increased UV and U(Na)V in the face of decreased pressure suggests a direct renal effect. PMID- 10685725 TI - Pressor responses to serotonin injected into the nucleus tractus solitarius of Sprague-Dawley rats and spontaneously hypertensive rats. AB - Previous studies in rats have shown that injection of nanomoles of serotonin (5 hydroxytryptamine; 5HT) into the nucleus tractus solitarius (NTS) acts on 5HT3 receptors to increase arterial pressure (AP). We investigated the effect of 5HT in Sprague-Dawley (SD) rats and in spontaneously hypertensive rats (SHR). Injection of nanomoles of 5HT into the NTS of chloralose-anesthetized SD rats increased AP. This effect was inhibited by prior injection of 5HT3 receptor antagonist ondansetron. The GABA(A) receptor antagonist bicuculline did not inhibit the effect of 5HT. Bilateral injection of 5HT or ondansetron did not affect the baroreflex sensitivity. Bilateral injection of ondansetron did not alter AP. The pressor effect of 5HT was exaggerated in SHR. These results suggest that stimulation of 5HT3 receptors in the NTS increases AP independently of activation of GABAA receptors and the baroreflex sensitivity. Furthermore, this serotonergic system is supersensitive in the NTS of SHR. PMID- 10685726 TI - Increased arginase activity in aorta of mineralocorticoid-salt hypertensive rats. AB - The present study was designed, first to investigate aortic arginase activity during the development and the establishment of mineralocorticoid-salt (DOCA salt) hypertension, and second, to determine the relationship between arginase activity and blood pressure by giving a protein-supplemented diet (50% casein) known to increase hepatic arginase activity. Our results showed that aortic arginase activity in established hypertension of DOCA-salt rats was higher than in normotensive rats. The protein-supplemented diet (50% casein) accelerated the development of DOCA-salt hypertension. There was a positive correlation between arginase activity and the level of blood pressure in these DOCA-salt hypertensive rats fed 50% casein but not in DOCA-salt hypertensive rats on a normal (20% casein) diet. In normotensive rats, the protein-supplemented diet decreased aortic arginase activity and produced no change in systolic blood pressure. Our data suggest that aortic arginase activity is modified in established DOCA-salt hypertension and could participate in the physiopathology of arterial hypertension. PMID- 10685727 TI - Renin-angiotensin-aldosterone system gene polymorphisms and hypertension in Hong Kong Chinese. AB - In Chinese populations, hypertension is common and is a major risk factor for cerebrovascular and coronary heart disease. The renin-angiotensin-aldosterone system (RAAS) helps maintain blood pressure and salt homeostasis and appears important in the pathogenesis of hypertension and some forms of vascular disease. We investigated three RAAS gene polymorphisms, the angiotensin-converting enzyme (ACE) insertion/deletion, angiotensinogen (AGT) M235T and angiotensin II type 1 receptor A1166C polymorphisms in 232 hypertensive and 178 normotensive Chinese subjects. The hypertensives were generally more obese and dyslipidaemic. No significant differences in genotype or allele frequencies for any of the polymorphisms were identified between the groups, nor was there any interactive contribution to blood pressure by the ACE and AGT polymorphisms. However, there were large differences in genotype and allele frequencies between the healthy Chinese and published data for equivalent Caucasian populations. These findings suggest these polymorphisms are unlikely to be involved in the pathogenesis of hypertension in Chinese. PMID- 10685728 TI - Possible relationship between altered fatty acid composition of serum, platelets, and aorta and hypertension induced by sugar feeding in rats. AB - To establish a relationship between alterations in fatty acid metabolism, induced by sugar ingestion, and hypertension, we analyzed fatty acid composition of serum, platelets and aorta in rats which had 30% of sugar in their drinking water for 18-20 weeks, and became hypertensive, hypertriglyceridemic and hyperinsulinemic. The fatty acid composition in sugar-fed as compared with that from control rats was as follows: in serum phospholipids, triglycerides and cholesterol ester fractions, palmitic, palmitoleic, oleic and cis-11 eicosadecaenoic acids were present in a higher proportion. In serum phospholipid fraction linoleic and arachidonic acids were decreased and a significant increase was observed in the proportion of dihomo-gamma-linolenic acid. In the membrane phospholipids of platelets and aorta, higher proportions of palmitoleic and of oleic acids were observed. Differences in fatty acid composition of phospholipids between sugar-fed and control rats are consistent with altered membrane fluidity. Altered membrane function is a potential mechanism involved hypertension in rats in sugar-induced. PMID- 10685729 TI - Ovariectomy alters the chronic hemodynamic and sympathetic effects of ethanol in radiotelemetered female rats. AB - This study determined the chronic hemodynamic effects of ethanol in telemetered freely moving female Sprague-Dawley rats. The role of ovarian hormones and sympathetic activity in the modulation of ethanol responses was also investigated. Changes in blood pressure (BP), heart rate (HR), and plasma estrogen and norepinephrine (NE, as index of sympathetic activity) were evaluated in pair-fed sham-operated (SO) and ovariectomized (OVX) rats receiving liquid diet with or without ethanol (5%, w/v) for 12 weeks. OVX caused a significant increase (about 40 g) in body weight, compared with the sham operation, which was apparent after two weeks and remained so for the duration of the study. The body weight showed gradual and similar increases in both ethanol and control groups. Ethanol feeding had no effect on the plasma estrogen level in SO or OVX rats. Daily ethanol intake was significantly (P < 0.05) less in OVX compared with SO rats whereas the blood ethanol concentration were similar in the two groups except for a significantly (P < 0.05) higher level in OVX rats at weeks 8, 10, and 11. Ethanol feeding caused significant (P < 0.05) decreases in BP in SO rats that started at week land reached maximal response (approximately 10 mmHg) at week 6 and remained at that level till the end of week 12. In OVX rats, ethanol had no effect on BPduring the first 5 weeks of the study. A slight but significant reduction in BP (about 5 mmHg) by ethanol in OVX rats started to appear at week 6 and remained for the following 5 weeks. The reduced hypotensive effect of ethanol in OVX rats was associated with an increase in the sympathetic activity as indicated by the significant (P < 0.05) increases in plasma NE levels. This sympathoexcitatory effect of ethanol was not demonstrated in SO rats. The HR was not affected by ethanol in the two groups of rats except for significant (P < 0.05) increases at weeks 1 through 3 in SO rats. The present findings suggest that the ovarian hormones modulate, at least partly, the hemodynamic and neurohumoral effects of chronic ethanol feeding in female rats. Ethanol lowers BP in female rats and this effect was delayed and diminished in OVX rats due possibly to the associated increase in sympathetic activity. PMID- 10685730 TI - Reflux esophagitis: healed! Now what? PMID- 10685731 TI - Public and physician education improves participation in colorectal cancer screening. PMID- 10685732 TI - Irritable bowel syndrome and health care seeking: do we pass our bad habits onto our children? PMID- 10685733 TI - A new weapon for the arsenal in the war against constipation? PMID- 10685734 TI - Scopes, hopes, and learning the ropes. PMID- 10685735 TI - Helping patients with Crohn's disease quit smoking. AB - Smoking is not only a risk factor for Crohn's disease, but ongoing smoking is associated with a poorer disease course. Therefore, smoking cessation should be an important treatment strategy for Crohn's disease patients who smoke tobacco. Recent improvements in understanding how people quit smoking and the development of pharmacological interventions, such as nicotine patches and bupropion, have improved cessation rates. In this article, we first briefly review the evidence supporting the adverse effects of smoking on the disease course. We next review the current understanding of how people change addictive behaviors, such as smoking. We then describe how the gastroenterologist can aid the patient with Crohn's disease to quit smoking, including appropriate and brief counseling strategies and the use of adjunctive treatments. Given the improvements in smoking cessation strategies, all patients with Crohn's disease should be strongly advised to quit smoking and be aided in doing so. PMID- 10685736 TI - Established and emerging biological activity markers of inflammatory bowel disease. AB - Assessment of disease activity in inflammatory bowel disease (IBD), i.e., ulcerative colitis (UC) and Crohn's disease (CD), is done using clinical parameters and various biological disease markers. Ideally, a disease marker must: be able to identify individuals at risk of a given disorder, be disease specific, mirror the disease activity and, finally, be easily applicable for routine clinical purposes. However, no such disease markers have yet been identified for IBD. In this article, classical disease markers including erythrocyte sedimentation rate, acute phase proteins (especially orosomucoid and CRP), leukocyte and platelet counts, albumin, neopterin, and beta2-microglobulin will be reviewed together with emerging disease markers such as antibodies of the ANCA/ASCA type, cytokines (e.g., IL-1, IL-2Ralpha, IL-6, IL-8, TNF-alpha, and TNF alpha receptors) and with various adhesion molecules. It is concluded that none of the pertinent laboratory surrogate markers of disease activity in IBD are specific or sensitive enough to replace basic clinical observation such as the number of daily bowel movements, general well-being, and other parameters in parallel. Further studies are highly warranted to identify and assess the clinical importance and applicability of new laboratory markers for the diagnosis or the disease activity of IBD. PMID- 10685737 TI - Gastroesophageal reflux disease in the older patient: presentation, treatment, and complications. AB - Gastroesophageal reflux disease (GERD) is common in the elderly. Patients often complain of less severe or frequent heartburn than their younger cohorts, but because of prolonged acid exposure over many years, the elderly have more complicated reflux disease including esophagitis, peptic strictures, and Barrett's esophagus. Potential factors aggravating GERD in the elderly include medications, which reduce lower esophageal sphincter pressure, higher frequency of hiatal hernia, impaired motility, and decreased saliva volume and bicarbonate concentration. Early endoscopy is indicated in all elderly patients with GERD, regardless of symptom severity. The medical and surgical treatment of GERD in the elderly generally follows the same principles as for any adult patient. PMID- 10685738 TI - Selection of patients for successful maintenance treatment of esophagitis with low-dose omeprazole: use of 24-hour gastric pH monitoring. AB - OBJECTIVE: Treating patients with erosive esophagitis and maintaining remission in a cost-effective fashion is a desirable goal in clinical practice. There are no established criteria to identify patients with healed esophagitis who will subsequently remain in remission with low-dose omeprazole therapy. We investigated whether 24-h esophageal-gastric pH monitoring could provide criteria to select patients for low-dose omeprazole maintenance therapy. METHODS: Seventy consecutive symptomatic outpatients with grade 2-3 reflux esophagitis were prospectively investigated. They were treated with 20 mg/day omeprazole for 2 months. Those with healed esophagitis were given alternate-evening 20-mg omeprazole maintenance therapy for 6 months. Clinical evaluation, endoscopy, and 24-h esophageal-gastric pH were done at the end of each treatment period. Results of pH studies of patients in remission were compared with those with endoscopically documented relapse of esophagitis. RESULTS: In 63/70 patient (intention-to-treat, 90%; 95% confidence interval [CI], 83-97%) esophagitis was healed at 2 months. During the 6-month maintenance period esophagitis remain healed in 28 (G1) (40%; 95% CI, 29-52%), but recurred in 32 patients (G2). During healing with omeprazole 20 mg/day the 24-h gastric pH was below 4 for <10% of the time in 96% of the patients, who subsequently remained in long-term remission with low-dose maintenance therapy (G1), but not in any patient with recurrence of esophagitis (G2). The 10% threshold value has a specificity of 1.00 and sensitivity of 0.96. CONCLUSIONS: The 24-h intragastric pH monitoring during 20 mg/day omeprazole therapy provides criteria by which to preselect patients with reflux esophagitis who will remain in remission with low-dose omeprazole therapy. PMID- 10685739 TI - Ultrasonographic evaluation of lansoprazole-induced improvement of submucosal injury in patients with gastroesophageal reflux. AB - OBJECTIVE: Endoscopic ultrasonographic (EUS) changes in gastroesophageal reflux disease (GERD) after treatment with proton pump inhibitor have been poorly evaluated. We conducted a randomized, double-blind 12-wk clinical trial to compare the EUS effects of lansoprazole to histamine H2-receptor antagonist therapy in GERD. METHODS: Seventeen patients with reflux-related symptoms received 40 mg of famotidine for 6 wk or 30 mg of lansoprazole for 6 wk followed by 40 mg of famotidine or 30 mg of lansoprazole for another 6 wk, respectively. Patients underwent EUS before and at 6 and 12 wk after treatment. RESULTS: Before treatment, a variable degree of wall thickening was noted on EUS in the lower esophagus, compared with 20 normal subjects. After 6 wk of therapy, esophageal wall was significantly thicker in the famotidine group compared with the lansoprazole group (p<0.01). Surprisingly, thickening of esophageal wall and abnormal architecture were also detected in endoscopically negative reflux disease. Lansoprazole was superior to famotidine in reducing the thickness of esophageal wall. CONCLUSIONS: EUS was very useful for evaluation of submucosal injury in patients with GERD. EUS showed that a 6-wk course of lansoprazole therapy reduced thickening of esophageal wall, which was resistant to histamine H2-receptor antagonist therapy. Our results also suggest that inflammatory damage to the submucosal and muscle layers of the lower esophagus is the underlying mechanism of heartburn and associated symptoms in patients with endoscopically negative reflux disease. PMID- 10685740 TI - Prospective evaluation of the prevalence of gastric Helicobacter pylori infection in patients with GERD, Barrett's esophagus, Barrett's dysplasia, and Barrett's adenocarcinoma. AB - OBJECTIVE: This study was undertaken to prospectively determine the prevalence of gastric H. pylori infection in Barrett's esophagus and Barrett's complicated by dysplasia or adenocarcinoma. METHODS: The prevalence of H. pylori was determined in Barrett's esophagus patients compared to a control population of patients with gastroesophageal reflux disease (GERD) only. All patients had a minimum of 10 gastric surveillance biopsies obtained. H. pylori colonization was determined upon the basis of hematoxylin and eosin and use of a modified Giemsa and or Steiner's silver stain of all gastric biopsy specimens. RESULTS: Two hundred and eighty-nine Barrett's patients and 217 GERD control patients were included in the study. H. pylori was found in 95/289 (32.9%) of the Barrett's patients, compared with 96/217 (44.2%) of the GERD controls (NS). Forty-seven of the Barrett's patients had low-grade dysplasia/indefinite dysplasia, 14 high-grade dysplasia, and 20 Barrett's adenocarcinoma. When Barrett's was subgrouped according to absence of dysplasia, and presence of low-grade dysplasia, high-grade dysplasia, or adenocarcinoma, H. pylori prevalence was found to be significantly less for patients with Barrett's high-grade dysplasia (14.3%) and adenocarcinoma (15.0%) versus patients with GERD alone (44.2%), Barrett's alone (35.1%), or Barrett's with low-grade dysplasia (36.2%) (p = 0.016). This difference could not be explained by differences between Barrett's esophagus patients infected with H. pylori and those who were not with respect to gender, smoking history, alcohol consumption, use of proton pump inhibitor, or length of Barrett's mucosa. CONCLUSIONS: Barrett's high-grade dysplasia and adenocarcinoma are significantly more prevalent in patients who are not infected with H. pylori. H. pylori appears to have a protective effect against the development of Barrett's adenocarcinoma. PMID- 10685741 TI - A cost-effectiveness analysis of prescribing strategies in the management of gastroesophageal reflux disease. AB - OBJECTIVE: Patients who have uncomplicated gastroesophageal-reflux disease (GERD) typically present with heartburn and acid regurgitation. We sought to determine the cost-effectiveness of H2-receptor antagonists (H2RAs) and proton-pump inhibitors (PPIs) as first-line empiric therapy for patients with typical symptoms of GERD. METHODS: Decision analysis comparing costs and benefits of empirical treatment with H2RAs and PPIs for patients presenting with typical GERD was employed. The six treatment arms in the model were: 1) Lifestyle therapy, including antacids; 2) H2RA therapy, with endoscopy performed if no response to H2RAs; 3) Step up (H2RA-PPI) Arm: H2RA followed by PPI therapy in the case of symptomatic failure; 4) Step down arm: PPI therapy followed by H2RA if symptomatic response to PPI, and antacid therapy if response to H2RA therapy; 5) PPI-on-demand therapy: 8 wk of treatment for symptomatic recurrence, with no more than three courses per year; and 6) PPI-continuous therapy. Measurements were lifetime costs, quality-adjusted life years (QALYs) gained, and incremental cost effectiveness. RESULTS: Initial therapy with PPIs followed by on-demand therapy was the most cost-effective approach, with a cost-effectiveness ratio of $20,934 per QALY gained for patients with moderate to severe GERD symptoms, and $37,923 for patients with mild GERD symptoms. This therapy was also associated with the greatest gain in discounted QALYs. The PPI-on-demand strategy was more effective and less costly than the H2RA followed by PPI strategy or the other treatment arms. The results were not highly sensitive to cost of therapy, QALY adjustment from GERD symptoms, or the success rate of the lifestyle arm. However, when the success rate of the PPI-on-demand arm was < or =59%, the H2RA-PPI arm was the preferred strategy. CONCLUSION: For patients with moderate to severe symptoms of GERD, initial treatment with PPIs followed by on-demand therapy is a cost effective approach. PMID- 10685742 TI - Predictability of dysphagia after laparoscopic nissen fundoplication. AB - OBJECTIVE: Dysphagia is the most common complication of antireflux surgery. Temporary dysphagia occurs in addition to persistent dysphagia because of technical or physiological problems. Temporary dysphagia may be due to the patient's personal perception or faulty eating habits. The aim of this study was to investigate the impact of the patient's personality as it relates to temporary dysphagia and individual impairment. METHODS: Several studies have used the construct of personality known as "health locus of control" to predict health related behavior and convalescence after medical or surgical treatments. This study investigates the predictability of the subjective degree of dysphagia and its perceived degree of impairment in relation to the health locus of control after laparoscopic so-called "floppy" Nissen fundoplication in 90 patients. Several questionnaires and single-item questions were given to the patients preoperatively, and 1 wk, 6 wk, and 3 months after surgery. The answers to the questions provided the data for this study. RESULTS: Preoperatively, 92% of the patients had no dysphagia and 8% had a mild subjective degree of dysphagia. Temporary postoperative dysphagia was found in approximately 50% of the patients 1 wk after surgery. The intensity of the dysphagia ranged among mild (18%), moderate (15%), and severe (16%). Three months postoperatively about 80% had no dysphagia and only 2% severe dysphagia. Correlations between the construct of personality and the intensity of postoperative dysphagia and its impairment revealed a significant relationship at all times. Patients with high expectations for their own health-related abilities (internal control) had less dysphagia (r = -0.78 after 1 wk [p<0.001], r = -0.71 after 6 wk [p<0.001], and r = -0.64 after 3 months [p<0.001]), compared with patients who believed that their convalescence depended more on luck, chance, or fate (external control) (r = 0.67 after 1 wk [p<0.01], r = 0.72 after 6 wk [p<0.001], and r = 0.63 after 3 months [p<0.01]). These results are highly significant. The correlation between health locus of control the degree of a subjective impairment from perceived dysphagia showed similar results (p<0.01). CONCLUSIONS: The subjective degree of dysphagia and the perceived impairment as a result of laparoscopic antireflux surgery can be predicted according to the personality of the patient. Those patients with low expectations for their own abilities can be identified before surgery, thereby allowing adaptation techniques to be applied that could improve the results and well-being of patients after antireflux surgery. PMID- 10685743 TI - Gastrointestinal bleeding in patients with hereditary hemorrhagic telangiectasia. AB - OBJECTIVE: Gastrointestinal bleeding occurs in a number of patients with hereditary hemorrhagic telangiectasia (HHT) and may lead to a high transfusion need. The aim of this study was to estimate the occurrence and severity of gastrointestinal bleeding in a geographically well defined HHT population. METHODS: All HHT families in the county of Fyn, Denmark, (470,000 population) have been identified. Probands and their first degree relatives, and all descendants from probands for whom one parent had HHT were eligible for inclusion in the study. A total of 77 patients with HHT were identified; of these, 76 patients (mean age: 52 yr) were evaluated and interviewed with regard to gastrointestinal bleeding, that is, a history of either hematemesis or melena. Patients charts were reviewed. RESULTS: A total of 25 HHT patients (33%) had a history of either hematemesis or melena. Of these, 12 (48%) had received blood transfusions. Seven patients had severe bleeding (that is, > or =6 units of blood within 6 months before inclusion in the study). Endoscopy had been performed in 16 of the 25 (64%) patients. Telangiectatic lesions were documented in nine at upper endoscopy and in one at sigmoidoscopy. Telangiectatic lesions were observed in all patients with severe bleeding, but in two patients epistaxis is likely to have contributed to the anemia. Among 51 HHT patients without a history of gastrointestinal bleeding, only five (10%) had previously received blood transfusions; however, none fulfilled the definition of severe bleeding. In the HHT population 29 patients were > or =60 yr old, but all patients with severe bleeding were > or =60 yr. CONCLUSIONS: A history of gastrointestinal bleeding is common in patients with HHT (33%). This study documents that 25% of HHT patients > or =60 yr suffer from severe gastrointestinal bleeding. PMID- 10685744 TI - Effects of postprandial walking on delayed gastric emptying and intragastric meal distribution in longstanding diabetics. AB - OBJECTIVE: We investigated the influence of standardized postprandial walking on the rates of gastric emptying and of intragastric meal distribution in 50 consecutive patients with longstanding insulin-dependent diabetes mellitus. METHODS: Gastric emptying of a semisolid meal labeled with 99mTc was continuously recorded with a dual-head gamma camera with patients in the supine position for 90 min before and 20 min after a 30-min postprandial walk. Regions of interest enclosing total stomach, and proximal and distal gastric compartments were calculated to determine gastric emptying rates and intragastric meal distribution. RESULTS: The evaluation of gastric emptying rates before and after postprandial walking demonstrated two variants of delayed gastric emptying: one variant that was counteracted by postprandial walking in seven patients (14%, Group I) and another variant that was not influenced by postprandial walking in 11 patients (22%, Group II). In addition, the emptying rates of 28 patients (56%) were within the range of controls and in four patients the emptying was accelerated (8%). The filling of the proximal gastric compartment was predominant and remained dominant after walking in Groups I and II. In controls and in diabetics with normal gastric emptying, the preliminary predominant filling of the proximal compartment was equalized after walking and the proximal compartment regained predominance thereafter. The changes in gastric emptying characteristics from delayed to accelerated gastric emptying may be related to the duration of diabetes (r = -0.47, p<0.03) and were not indicated by symptoms of upper GI discomfort or by secondary diabetic manifestations. CONCLUSION: Postprandial walking may improve gastric emptying in 14% of patients with longstanding insulin dependent diabetes mellitus. PMID- 10685745 TI - Enhanced postprandial gastric myoelectrical activity after moderate-intensity exercise. AB - OBJECTIVE: The aim of this study was to investigate the change of postprandial gastric myoelectrical activity and its relation with vagal activity after exercise. METHODS: Nine subjects were studied in two sessions. In the control session, gastric myoelectrical activity was recorded using electrogastrography (EGG) for 30 min in the fasting state and 60 min after a test meal. In the exercise session, after the baseline recording of both the EGG and electrocardiogram (ECG), the subject was put on a cycle ergometer for exercise until reaching 50% of the maximum age-predicted heart rate for 10 min. The test meal was then given and the recording was resumed for 60 more minutes. Spectral analyses were performed on both the EGG and the heart rate variability derived from the ECG. RESULTS: The postprandial increment of the dominant power (p<0.05) and the percentage of the 2-4 cpm slow waves (p = 0.01) were significantly higher with exercise. The standard deviation of the postprandial dominant frequency was significantly decreased (more stable slow waves) with exercise (p<0.04). While cardiac vagal activity was significantly decreased after the meal, exercise did not significantly affect the postprandial change. CONCLUSIONS: Gastric slow waves become more regular, more stable, and of higher amplitude after exercise, and this enhancement is probably not mediated via the vagal pathway. PMID- 10685746 TI - Malignant perihilar biliary obstruction: magnetic resonance cholangiopancreatographic findings. AB - OBJECTIVE: We studied the efficacy of magnetic resonance cholangiopancreatography (MRCP) in the evaluation of malignant perihilar biliary obstructions, with reference to endoscopic retrograde cholangiopancreatography (ERCP). METHODS: A total of 40 patients with malignant perihilar biliary obstructions, who underwent both MRCP (Magnetom Vision; Siemens, Erlangen, Germany; projection technique and multislice plus maximum intensity projection) and ERCP examinations, were studied. The study group included hilar cholangiocarcinoma (Klatskin tumor) in 26 patients, icteric hepatocellular carcinoma in four patients, gallbladder carcinoma in five patients, and metastasis from other than hepatobiliary origin in five patients. Axial and coronal magnetic resonance (MR) images were added simultaneously to the MRCP. The mean serum bilirubin level on admission was 11.5 mg/ml (range, 2.8-28.5 mg/ml). The presence and extent of malignant biliary obstruction were determined with both MRCP and ERCP following the known criteria: an abrupt and irregular character of a distal narrow segment, a proportionally dilated biliary tree proximally, and an irregularly shaped intraluminal filling defect. The efficacy of the MRCP examination in detecting the presence of biliary obstruction, its anatomical extent, and the underlying cause, respectively, was compared to that of ERCP. RESULTS: MRCP examination was successfully performed on all patients, whereas ERCP examination was unsuccessful in two patients. Both MRCP and ERCP were very effective in detecting the presence of biliary obstructions (40 of 40 vs. 38 of 38, p = 1.0). MRCP was superior in its investigation of anatomical extent (34 of 40 vs. 24 of 38, p = 0.015) and the cause of the jaundice (31 of 40 vs. 22 of 38, p = 0.023) compared to ERCP. Specifically, the performance of MRCP is promising for the interpretation of cholangiocarcinoma (22 of 26) and gallbladder carcinoma (five of five), but is relatively ineffective for the interpretation of icteric HCC (two of four) and metastasis (two of five). CONCLUSION: MRCP represented an ideal noninvasive diagnostic tool for the evaluation of malignant perihilar biliary obstructions with reference to ERCP. PMID- 10685747 TI - Outcome after surgical resection of intraductal papillary and mucinous tumors of the pancreas. AB - OBJECTIVE: Treatment of intraductal papillary and mucinous tumors of pancreas (IPMT) usually requires surgery. The objective of this study was to evaluate the risk of recurrence in patients after surgery according to the histological nature of the neoplasm and the type of surgery. METHODS: The outcome of 45 patients who underwent partial pancreatectomy (n = 35) or total pancreatectomy (n = 10) for IPMT was studied according to the nature of the neoplasm (invasive carcinoma or noninvasive neoplasm), type of surgery (partial or total pancreatectomy), and lymph nodes status. RESULTS: The overall 3-yr actuarial survival rate was 83%. Death occurred in seven of 20 (35%) patients with invasive carcinoma and in one of 26 (4%) patients with noninvasive tumors (p<0.05). There were two recurrences in the seven patients with noninvasive neoplasm who underwent partial pancreatectomy with involved resection margins, and none in the 13 patients with disease-free margins. In patients with invasive carcinoma, there was one recurrence after total pancreatectomy, six after partial pancreatectomy with disease-free margins and six after partial pancreatectomy with involved margins. In patients with invasive carcinoma, total pancreatectomy and the absence of lymph nodes involvement were independently associated with a low risk of recurrence. CONCLUSIONS: IPMT may be managed as follows: 1) in patients with noninvasive neoplasms, partial pancreatic resection should be guided by frozen section examination until disease-free margins are obtained; and 2) in patients with invasive carcinoma, total pancreatectomy seems most likely to cure the patient, but should be discussed according to the general status and the age. PMID- 10685748 TI - A randomized, placebo-controlled, multicenter study of the safety and efficacy of a new polyethylene glycol laxative. AB - OBJECTIVE: This study was designed to determine the efficacy and safety of a new laxative, Braintree polyethylene glycol (PEG) laxative (Miralax, Braintree Laboratories, Braintree, MA). METHODS: This investigation was designed as a placebo-controlled, blinded, randomized, multicenter parallel trial. Study subjects were constipated but otherwise healthy outpatients who had < or =2 stools during a 7-day qualification period. Braintree PEG laxative 17 g or dextrose placebo p.o. in 8 oz of water for a 14-day treatment period. A diary recorded each bowel movement and subjective symptoms of stool consistency, ease of passage, cramps, and flatus. CBC, blood chemistries and urinalysis were performed before and after the treatment period. RESULTS: There were 151 randomized subjects, 131 female and 20 male. An increase in bowel movement frequency was observed with the PEG laxative as compared to placebo (p<0.001), with the greatest difference in efficacy in wk 2 of treatment (p<0.001). By wk 2 of treatment, on average, placebo subjects had 2.7 bowel movements/wk and PEG treated study subjects had 4.5 movements/wk (p<0.01), or more than one bowel movement every 2 days. Investigator (p<0.005) and patient (p<0.001) subjective assessment of perception of treatment effectiveness, and patient evaluations of stool consistency and passage showed significant improvement in the active treatment group (p<0.001). There were no significant differences in laboratory changes or adverse experiences recorded between groups. CONCLUSION: Braintree PEG laxative is safe and effective in the short term for the treatment of constipation. PMID- 10685749 TI - Intergenerational transmission of gastrointestinal illness behavior. AB - OBJECTIVE: Previous research, based on retrospective reporting, suggests that parental reinforcement and modeling may be important mechanisms in the development of gastrointestinal illness behavior in children and adults. The aim of this study was to determine the relationship between the illness behavior of parents, in the form of health care use for irritable bowel symptoms, and the illness behavior of their children, without relying on retrospective recall. METHODS: A comparison of two matched groups was made. Groups included 631 children of parents who were diagnosed with irritable bowel syndrome during 1 calendar yr and 646 children of parents matched by parental age, gender, and number of children in the family who did not receive an IBS diagnosis during the same 1 yr. Health care use and costs over a 3-yr calendar period for all children and their parents collected from the health care database of a large health maintenance organization were evaluated. RESULTS: Case children had significantly more ambulatory care visits for all causes (mean 12.26 vs. 9.81, p = 0.0001) and more ambulatory visits for gastrointestinal symptoms (0.35 vs. 0.18, p = 0.0001). Outpatient health care costs over the 3-yr period were also significantly higher for case than control children ($1979 vs. $1546, p = 0.0001). Controlling for the total number of ambulatory visits of the parents, excluding gastrointestinal visits, did not alter the findings. Gender of the IBS parent was not related to children's gastrointestinal visits. CONCLUSION: This study extends previous research by showing that specific types of illness behavior may be learned through modeling. PMID- 10685750 TI - Declining prevalence of opportunistic gastrointestinal disease in the era of combination antiretroviral therapy. AB - OBJECTIVE: Opportunistic disorders (OD) are the most frequent GI manifestations of the acquired immunodeficiency syndrome (AIDS). Since the introduction of highly active antiretroviral therapy (HAART), there appears to be have been a reduction in the incidence of many of these OD; however, the effect of HAART on the prevalence of GI OD has not been well studied. METHODS: From 4/95 through 3/98, all HIV (HIV)-infected patients undergoing GI endoscopy were prospectively identified; mucosal biopsies were obtained in a standardized fashion and histological specimens were examined by a single GI pathologist. Patients were divided into three groups based on the time of evaluation: group I: 4/95 to 3/96; group II: 4/96 to 3/97; and group III: 4/97 to 3/98. RESULTS: A total of 166 patients (90% men; mean age 36+/-10 yr; median CD4 lymphocyte count 62 cells/microl, range 2-884, median viral RNA level 1,357 copies/ml, range undetectable to 7,721,715) underwent 279 upper and/or lower endoscopies during the study period. There were no statistical differences in patients' demographics and indications for endoscopy although the CD 4 lymphocyte count was higher in group III. The percentage of patients receiving HAART at the time of endoscopy increased from 0% to 57% over the three periods (p<0.01), and the percentage of patient receiving combination antiretroviral therapy increased from 37% to 82% over the study period (p<0.01). In contrast, the prevalence of OD decreased from 69% (group I) to 13% (group III) (p<0.01), whereas the prevalence of non-OD, including a normal endoscopy increased from 31% to 87% (p<0.01). CONCLUSIONS: GI OD now seem to be an uncommon problem in HIV-infected patients undergoing endoscopy despite a low CD4 lymphocyte count, and this reduction of OD was associated with the use of HAART. PMID- 10685751 TI - The natural history of gluten sensitivity: report of two new celiac disease patients resulting from a long-term follow-up of nonatrophic, first-degree relatives. AB - OBJECTIVE: Early studies revealed that up to 50% of non-atrophic, first-degree relatives of celiac disease patients exhibit features of gluten sensitivity. However, whether these features progress to a fully expressed celiac disease remain partially known. Our aim was to report two new patients resulting from a prospective, long-term surveillance of relatives who were nonatrophic at initial assessment. METHODS: After a median time of 86 months (range: 42-102 months) from the baseline assessment, we re-evaluated 44 first-degree relatives of propositi who had taken part in family studies and in whom baseline small intestinal biopsies were normal. At the baseline screening, 21 relatives had positive serum antigliadin antibodies and/or increased intraepithelial lymphocyte infiltration, and 23 did not. In addition, 11 of 18 had a celiac-like response to rectal gluten challenge and 16 of 34 possessed the characteristic HLA DQ2 haplotype (DQA1 0501 DQB1 0201). Re-evaluation was based on celiac-related serology antigliadin (AGA) and endomysial (EmA) antibodies. EmA-positive subjects underwent intestinal biopsy. RESULTS: At the end of the study, EmA was positive in only two subjects. Histological examination revealed flat small bowel mucosa in both. At baseline, both cases were EmA-negative and no minor histological changes were observed. One was a woman with positive baseline IgA and IgG AGA and a rectal gluten challenge with a celiac-like response; the other patient has presented only with a positive IgG AGA. In both cases, progression was detected in a clinically silent context. Both new patients had the characteristic HLA DQ2 haplotype. CONCLUSIONS: Our data suggest the need to re-evaluate relatives who have been negative on initial screening for celiac disease. Up to now, the progression to severe enteropathy was only observed in relatives who had presented some evidence of gluten sensitivity and the characteristic HLA DQ2 haplotype. Longer longitudinal studies are necessary to obtain definitive conclusions. PMID- 10685752 TI - The natural history of ulcerative proctitis: a multicenter, retrospective study. Gruppo di Studio per le Malattie Infiammatorie Intestinali (GSMII). AB - OBJECTIVE: The aim of this study was to evaluate the clinical features and the long term evolution of patients with a well defined initial diagnosis of ulcerative proctitis. METHODS: Patients with an original diagnosis of ulcerative proctitis who had been seen at any of 13 institutions from 1989 to 1994 were identified. Data on disease onset and subsequent evolution were recorded. In addition, 575 patients with more extensive disease, treated in the same centers, were used as controls. RESULTS: A total of 341 patients satisfied the inclusion criteria. The percentage of smokers in these patients was slightly lower than in controls; no differences were found in the other clinical/demographic variables evaluated. A total of 273 patients entered long term follow-up (mean, 52 months). Proximal extension of the disease occurred in 74 of them (27.1%). The cumulative rate of proximal extension and of extension beyond the splenic flexure was 20% and 4% at 5 yr and 54% and 10% at 10 yr, respectively. The risk of proximal extension was higher in nonsmokers, in patients with >3 relapses/yr, and in patients needing systemic steroid or immunosuppressive treatment. Refractory disease was confirmed as an independent prognostic factor at multivariate analysis. CONCLUSIONS: Proximal extension of ulcerative proctitis is frequent and may occur even late after the original diagnosis. However, the risk of extension beyond the splenic flexure appears to be quite low. Smoking seems to be a protective factor against proximal extension, whereas refractoriness is a risk factor for proximal extension of the disease. PMID- 10685753 TI - Lack of association between smoking and Crohn's disease but the usual association with ulcerative colitis in Jewish patients in Israel: a multicenter study. AB - OBJECTIVE: The association between smoking and inflammatory bowel disease (IBD) is well established, but data in Jewish patients in Israel were discrepant. The aim of this study was to examine the smoking habits of Jewish IBD patients in Israel in a large scale, multicenter study. METHODS: Patients with established IBD aged 18-70 yr were interviewed in relation to smoking and other habits. Two controls (one clinic and one neighborhood control matched by age, sex, community group, and education) were sought for each subject. RESULTS: A total of 534 patients (273 ulcerative colitis [UC], and 261 Crohn's disease [CD]), along with 478 clinic controls and 430 neighborhood controls, were interviewed. There was no significant difference in the smoking habits between CD patients and their controls. Of patients with CD, 24.5% were current smokers, as compared to 19.9% of clinic controls and 25.2% of neighborhood controls (NS). The odds ratio for CD in current smokers was 1.30 (95% confidence interval 0.85-1.99) versus clinic controls, and 0.96 (0.63-1.46) versus neighborhood controls. There were also no significant differences in the proportion of ex-smokers between the groups. Only 12.9% of UC patients were current smokers versus 21.9. % Clinic controls, and 26.4% community controls (p<0.005). The proportions of ex-smokers were higher in UC patients 29.7% versus 25.9%, and 19.5% in their respective controls (p<0.001 vs. community controls). No significant differences were found in the proportions of never-smokers between IBD patients and controls. All the above trends were similar in four different parts of the country. The proportion of current smokers in UC decreased with the extent of disease (19.7% in proctitis, 13.6% in left sided, and 4.5% in total colitis) (p<0.05). Patients with UC were more likely to be light smokers(1-10 cigarettes/day), whereas patients with CD were more likely to be moderate smokers (11-20 cigarettes/day) in comparison to their controls. CONCLUSIONS: The lack of association between smoking and CD has now been established in Jewish patients in Israel. The association was found in UC. The stronger genetic tendency in CD may contribute to this discrepancy. PMID- 10685754 TI - Crohn's disease: does race matter? The Mid-Atlantic Crohn's Disease Study Group. AB - OBJECTIVE: The severity of Crohn's disease (CD) has been reported to be greater in blacks than in whites. This possible disparity may be due, in part, to differences between these groups in health care utilization and accessibility. To explore these issues, we conducted a multicenter survey of patients with CD. METHODS: One-hundred and forty-five blacks with CD, recruited from four teaching hospitals and five private practices, and identified by medical record review or ICD-9 code, were enrolled and matched to 407 whites with CD (by age, gender, and practice type [teaching vs. private practice setting]). Participants were interviewed regarding medical history, health status, personal health care practices during the preceding 5 yr, and beliefs regarding health care in the general population. RESULTS: Blacks and whites were similar with respect to age of CD onset, lag in time to diagnosis, and number of gastrointestinal (GI) related hospitalizations and surgeries. Medication usage patterns were also similar in the two groups. Quality of life, measured by SF-36, was lower in all categories for blacks, compared with whites. Blacks were more likely to have had to stop work (p<0.01) and have lost more work days (p<0.01) than were whites. Whites were more likely to have health insurance and be able to identify a regular provider than were blacks. Blacks were more likely to report the following: receiving Medicaid; difficulty affording health care; delaying appointments due to financial concerns; difficulty traveling to their provider's office; and experiencing unreasonable delays at their provider's office. After adjusting for potential confounding variables, we found no differences between the groups, except for the number of days of work lost because of CD. CONCLUSIONS: These data suggest that black and white patients have similar reported disease presentations and course, and contrast with prior reports suggesting a more severe disease course among black patients. Although the disease itself appears similar, there were numerous reported differences between the races in health care utilization practices and in disease impact upon daily activities. We suggest that apparent disparities in CD according to race are actually due to social and economic factors, and not to the disease itself. PMID- 10685755 TI - Upper gastrointestinal bleeding in patients with hepatic cirrhosis: clinical course and mortality prediction. AB - OBJECTIVE: We conducted this study to describe the complications and validate the accuracy of previously reported prognostic indices in predicting the mortality of cirrhotic patients hospitalized for upper GI bleeding. METHODS: This prospective, observational study included 111 consecutive hospitalizations of 85 cirrhotic patients admitted for GI bleeding. Data obtained included intensive care unit (ICU) admission status, Child-Pugh score, the development of systemic inflammatory response syndrome (SIRS), organ failure, and inhospital mortality. The performances of Garden's, Gatta's, and Acute Physiology and Chronic Health Evaluation (APACHE) II prognostic systems in predicting mortality were assessed. RESULTS: Patients' mean age was 48.7 yr, and the median APACHE II and Child-Pugh scores were 17 and 9, respectively. Their ICU admission rate was 71%. Organ failure developed in 57%, and SIRS in 46% of the patients. Nine patients had acute respiratory distress syndrome, and three patients had hepatorenal syndrome. The inhospital mortality was 21%. The APACHE II, Garden's, and Gatta' s predicted mortality rates were 39%, 24%, and 20%, respectively, and their areas under the receiver operating characteristic curve (AUC) were 0.78, 0.70, and 0.71, respectively. The AUC for Child-Pugh score was 0.76. CONCLUSIONS: SIRS and organ failure develop in many patients with hepatic cirrhosis hospitalized for upper GI bleeding, and are associated with increased mortality. Although the APACHE II prognostic system overestimated the mortality of these patients, the receiver operating characteristic curves did not show significant differences between the various prognostic systems. PMID- 10685756 TI - Maintenance hemodialysis decreases serum hepatitis C virus (HCV) RNA levels in hemodialysis patients with chronic HCV infection. AB - OBJECTIVE: Hepatitis C virus (HCV) infection is a major complication among hemodialysis patients the world over. To determine the natural course of HCV viremic levels in patients on maintenance hemodialysis, we prospectively quantified the HCV RNA levels in serial blood samples from hemodialysis patients and compared them with those in nonuremic subjects. METHODS: The population studied included 98 hemodialysis patients and 228 nonuremic subjects with chronic HCV infection. HCV RNA was detected by polymerase chain reaction (PCR) and the levels were determined by branched DNA probe assay. HCV RNA genotypes were determined by PCR using type-specific primers. RESULTS: HCV RNA levels were significantly lower in hemodialysis patients (median, 0.4x10(6) genome equivalent [Meq]/ml) than in nonuremic subjects (median, 3.0 Meq/ml) (p<0.05). HCV of genotype 1b was prevalent in the hemodialysis patients (81.6%) and nonuremic subjects (88.6%). HCV RNA levels in 20 hemodialysis patients with genotype 1b were significantly reduced after each hemodialysis procedure (p<0.05). The 3-yr prospective observation from 1995 to 1998 showed a significant decrease of HCV RNA levels in 47 hemodialysis patients with genotype 1b (median, 1.9-0.9 Meq/ml, p<0.05), whereas levels in 155 nonuremic subjects with genotype 1b did not decrease (median, 2.6-3.0 Meq/ml). There were no patients or nonuremic subjects with undetectable HCV RNA by a PCR assay during the observation period. CONCLUSIONS: These observations suggest that maintenance hemodialysis decreases the HCV RNA levels in hemodialysis patients with chronic HCV infection, but does not produce clearance of the viremia. PMID- 10685757 TI - Cholestatic liver diseases and health-related quality of life. AB - OBJECTIVE: Symptoms associated with primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) negatively affect health-related quality of life (HRQL). The aim of this study was to measure HRQL in patients with chronic cholestatic liver diseases and to determine factors associated with more severe impairment. METHODS: We conducted a cross-sectional study in which we documented patients' demographic and clinical characteristics, and measured their HRQL using the Short Form-36 and Chronic Liver Disease Questionnaire. We assessed the association of HRQL impairment with disease severity (Child's-Pugh class and Mayo PBC Risk Score) and compared patients' HRQL with those of a healthy population, and patients with congestive heart failure, chronic obstructive pulmonary disease, and diabetes. RESULTS: One hundred and four patients with PBC and PSC participated, of whom 73% were women, with an average age of 55+/-12 yr. Of these patients, 61% had cirrhosis (37% Child's A, 23% Child's B, and 2% Child's C). Patients with cholestatic liver disease showed more HRQL impairment than the healthy population and were similar to patients with other chronic conditions. Additionally, patients who experienced severe itching showed profound HRQL impairment. In patients with PBC, Physical Component Summary (PCS) scores of the SF-36 and Chronic Liver Disease Questionnaire (CLDQ) scores fell from noncirrhotic to Child's A to Child's B/C and with worsening Mayo PBC Risk Scores. No other clinicodemographic data were associated with patients' well-being. CONCLUSIONS: Patients with cholestatic liver disease (PBC and PSC) showed substantial impairment of HRQL, which is further affected by worsening disease severity. Disease-specific measures were better able to discriminate patients with varying severities. PMID- 10685758 TI - Natural history of cirrhotic patients with small esophageal varices: a prospective study. AB - OBJECTIVE: Contrasting data are available on the natural history and bleeding risk of small esophageal varices. The aim of this prospective study was to evaluate a large series of consecutive cirrhotics with a first endoscopic diagnosis of small varices. METHODS: Between 1987 and 1992, 258 patients with small varices and no previous bleeding were enrolled. Patients were clinically examined every 6 months and were followed until a first episode of bleeding and/or death, or until June 1998. None received any treatment to prevent bleeding. Endoscopies were planned at 18-month intervals. RESULTS: The cumulative risk of bleeding was low (3% at 2 yr and 8% at 4 yr) and remained low in patients in whom varices remained small at 2nd endoscopy, whereas it increased significantly when varices enlarged. The increase of varices appeared to be rather linear in time: at the 2nd endoscopy varices remained small in 79% of patients and increased in 21%; at the 3rd endoscopy varices remained small in 55%, whereas at the 4th 33% of patients still had small varices. Clinical and biochemical data at the 1st and 2nd endoscopy were included in a multiple logistic regression analysis. Only the increase in Child-Pugh score appeared to be a significant predictor of enlarged varices; the risk of aggravation increased by 37.5% for every unit of impairment of the score. CONCLUSIONS: The present study shows that patients with small varices have a low bleeding risk. An increase in Child-Pugh score during follow-up suggests enlargement of varices, thus an increase in bleeding risk. In these patients closer endoscopic surveillance is recommended. PMID- 10685759 TI - Education improves colorectal cancer screening by flexible sigmoidoscopy in an inner city population. AB - OBJECTIVE: The District of Columbia General Hospital has a flexible sigmoidoscopy (FS) colorectal cancer screening program. We noted that this program was underused. The aim of this study was to determine whether education could improve use of a flexible sigmoidoscopy screening program in an inner city population. METHODS: Patients undergoing screening FS 5 months before our educational initiative were compared to patients undergoing screening FS 5 months after implementation. A 1-month period was allowed for implementation. Procedure logs and GI charts were reviewed. RESULTS: A total of 121 patients underwent FS screening during our study period. Of the patients, 97% were African-American; 58% were female; and the average age was 61 yr. A total of 50 patients underwent FS in the pre-education group, and 71 patients underwent FS after implementation of our educational initiative. CONCLUSIONS: Education resulted in a 42% increase in FS screening in this inner city, predominantly African-American population. Larger scale educational initiatives should be conducted to determine whether these benefits can persist and can be improved upon. PMID- 10685760 TI - Study of outcome after targeted intervention for peptic ulcer resistant to acid suppression therapy. AB - OBJECTIVE: Different factors might affect outcome in ulcers resistant to antisecretory therapy. The aim of the study was to define the odds of resistant ulcers being associated with NSAID use, and/or Helicobacter pylori (H. pylori) infection, or neither. METHODS: A total of 80 patients with resistant peptic ulcers were prospectively followed after targeted intervention for a mean follow up of 39.5+/-6.9 months. RESULTS: NSAID use was involved in 24 cases (14 with and 10 without concomitant H. pylori infection), H. pylori alone was involved in 44, and 12 patients had neither factor present. Of the NSAID group, resistant ulcers healed in patients who stopped taking NSAIDs. Those continuing to use NSAIDs (10 of 24; 41.6%) had either persistent ulceration or ulcer complications despite H. pylori eradication and omeprazole therapy. Of the H. pylori group, infection eradication induced ulcer remission in most patients, but those with persistent infection and a small subset of H. pylori eradicated patients (16.6%) had persistent/recurrent ulceration. Of the 12 refractory patients with neither NSAID use nor H. pylori infection, three had persistent ulceration but nine were controlled with antisecretory agents. Other factors (e.g., smoking or acid hypersecretion) were not associated with final outcome after targeted intervention of H. pylori infection and NSAID use. CONCLUSIONS: With current antiulcer therapies, NSAID use is the main, but not the exclusive, factor leading to intractability and complications in refractory ulcers. In a subset of resistant ulcers, neither the presence of H. pylori nor use of NSAIDs are involved. In this study, despite specific therapeutic intervention, 22.5% of patients with resistant ulcers had continuing ulcer problems. PMID- 10685761 TI - Stromal eosinophilia in colonic epithelial neoplasms. AB - OBJECTIVE: The purpose of this retrospective study was to determine the frequency and intensity of eosinophilic infiltration (or tissue eosinophilia) in the stroma of colonic adenomas, hyperplastic polyps, and colorectal adenocarcinomas. Eosinophilic infiltration in various malignancies has been reported but has not been evaluated in benign colorectal adenomas and hyperplastic polyps. METHODS: We analyzed 488 colonic neoplasms: 176 tubular adenomas, 55 tubulovillous adenomas, 82 villous adenomas, 15 early carcinomas in polyps, 95 invasive adenocarcinomas, and 65 hyperplastic polyps for the presence of eosinophilic infiltration. The eosinophilic infiltration was graded as negative (< or =5%), mild to moderate (>5 40%), or marked (>40%), depending on the percentage of eosinophils relative to total inflammatory cells in the stroma. RESULTS: Mild to moderate eosinophilia was noted in 75% of all adenomas. The transitional zone in all cases of invasive adenocarcinoma (zone between normal tissue and adenocarcinoma) revealed a high percentage of tissue eosinophilia. There was a striking absence of TE in the stroma of invasive adenocarcinomas. Only 5% of hyperplastic polyps had any eosinophilic infiltration. CONCLUSIONS: These data suggest that, in the spectrum of colonic neoplasms, stromal eosinophilia is most prominent in adenomas and seems to decrease with progression through the adenoma-carcinoma sequence. The ramifications of this study may alter management plans and provide some prognostic information for clinical evaluation. PMID- 10685762 TI - The cost of hospitalization in Crohn's disease. AB - OBJECTIVE: The aim of this study was to evaluate the demographics, resource use, and costs associated with hospitalization of Crohn's disease patients. METHODS: All patients hospitalized at our institution from 7/1/96 to 6/30/97 with a primary diagnosis of "Crohn's Disease" were analyzed using a computerized database. Data are presented "per hospitalization." RESULTS: A total of 175 hospitalizations (147 patients) were identified. Mean patient age was 36.5 yr; 61% were female; 82% Caucasian. Payer mix was most commonly contracted (57%), commercial (21%), or Medicare (13%). 57% of hospitalizations had a primary surgical procedure; the remainder were medical. Average length of stay was 8.7 days (surgical, 9.6 days; medical, 7.5 days). The average cost of hospitalization, excluding physician fees, was $12,528 (surgical, $14,409; medical, $10,020), whereas average charges were $35,378 (surgical, $46,354; medical, $20,744), including physician fees, which averaged $7,249 (surgical, $11,217; medical, $1,959). Mean reimbursements were $21,968 (surgical, $28,946; medical, $12,666) with average weighted reimbursement rates of 60.17% of hospital charges, 69.57% of physician fees. The distribution of costs across subcategories was: Surgery (39.6%), Pharmacy (18.6%), Laboratory (3.8%), Radiology (2.1%), Pathology (0.8%), Endoscopy (0.3%), and Other Hospital Costs (34.9%). Of the hospitalizations, 87% included treatment with steroids, 23% with immunomodulators, and 14% with aminosalicylates; 27% included the administration of total parenteral nutrition, which accounted for 63% of the total pharmacy costs. CONCLUSIONS: Surgery accounts for the majority of hospitalizations, nearly 40% of their total costs, and 75% of overall charges and reimbursements. Therapy that decreases the number of surgical hospitalizations should substantially reduce inpatient Crohn's disease costs, as well as overall costs. PMID- 10685763 TI - Trazodone-induced hepatotoxicity: a case report with comments on drug-induced hepatotoxicity. AB - Trazodone (Desyrel) is a second-generation, nontricyclic antidepressant that has been in use in North America since the early 1980s. It has the advantage of being more sedating and having less anticholinergic side effects than other secondary amines in the piperazine class, namely, desipramine and nortriptyline. Five previous cases of trazodone hepatotoxicity have been reported in the literature, one describing chronic damage and the others, more acute cellular and cholestatic injury. We describe a case of acute reversible liver injury with the use of trazodone. This case is unique in that injury occurred after protracted (18 months) drug use and while the patient was on corticosteroids. Moreover, the diagnosis was confirmed by an inadvertent challenge with trazodone. This case reports not only a well documented instance of trazodone-induced liver injury, but also serves as a basis for a brief discussion of mechanisms, clinical monitoring, and therapy in drug-induced hepatotoxicity. PMID- 10685764 TI - Duodenal mucosa-associated lymphoid tissue lymphoma: treatment with oral cyclophosphamide. AB - Small cell mucosa-associated lymphoid tissue (MALT) lymphomas rarely affect the duodenum, and optimal treatment has not been defined. The aim of this case series was to determine the clinical features and outcome of duodenal MALT lymphoma in four patients (three men, one woman; median age 52 yr) treated with cyclophosphamide p.o. Initial manifestations were abdominal pain (n = 4), vomiting (n = 2), and an obstructive syndrome (n = 1). MALT lymphoma was diagnosed on the basis of endoscopic biopsies. It was localized in the duodenum in three cases and involved the entire small bowel in one case. Tumor infiltration was limited to the duodenal wall in one case and was associated with locoregional lymphadenopathy in three cases. The patients were graded EI (n = 1) and EII1 (n = 3), respectively, according to the Ann Arbor classification revised by Musshof. Cyclophosphamide, 100 mg daily, was administered p.o. for 18 months. Gastroscopy with biopsies, radiography of the small intestine and abdominal CT (CT) were performed every 6 months. Complete remission was defined by morphological and histological normalization, and partial remission as morphological normalization only. Follow-up lasted from 9 to 65 months. Three patients were in complete remission at 18 months: two relapsed after 2 yr and one was still in complete remission at 65 months. The patient with 9 months of follow up was in complete remission at 6 months. The two patients who relapsed did not complain of symptoms, and no morphological abnormalities were seen. Relapse was diagnosed on histological grounds. Cyclophosphamide monotherapy p.o. thus seems well adapted to this slowly progressive disease, but it is unclear whether it should be resumed in the case of histological relapse or only in the case of symptomatic relapse. (Am J Gastroenterol 2000;95:536-539. (O 2000 by Am. Coll. of Gastroenterology) PMID- 10685765 TI - An endoscopic injection with N-butyl-2-cyanoacrylate used for colonic variceal bleeding: a case report and review of the literature. AB - We report a 64-yr-old patient with liver cirrhosis and bleeding esophageal varices that were obliterated by repeated endoscopic sclerotherapy. Eleven years later, he developed a massive, life-threatening rectosigmoid variceal hemorrhage. An endoscopic injection with N-butyl-2-cyanoacrylate (Histoacryl), performed over the rectosigmoid varices, achieved temporary hemostasis. The etiology, prevalence, relationship with portal hypertension, diagnosis, and treatment of colorectal varices are discussed. PMID- 10685766 TI - Life-threatening gastrointestinal hemorrhage due to juvenile polyposis. AB - A 14-yr-old, previously healthy boy presented with massive lower GI hemorrhage. After the routine endoscopic and radiological evaluation, laparotomy and intraoperative colonoscopy revealed multiple polyps in the colon. A hemicolectomy was performed because of the severity of hemorrhage. A diagnosis of juvenile polyposis was made based upon histological findings and the family history. This is an extremely unusual presentation of juvenile polyposis and has been reported only once before. The clinical features, diagnosis, and therapeutic options for juvenile polyposis are discussed. Juvenile polyposis, although a rare condition in the pediatric population, should be considered in the differential diagnosis of life-threatening GI hemorrhage. PMID- 10685767 TI - Stool testing for Helicobacter pylori infection: yet another noninvasive alternative. AB - A prospective, multicenter study was performed in 11 European centers to evaluate the accuracy of stool testing for active Helicobacter pylori (H. pylori) infection, using the HpSA ELISA antigen assay. The accuracy of this test was assessed in a large number of patients both before and after treatment by comparing results with a rigidly defined gold standard consisting of gastric antral and body biopsies with normal and special stains, as well as culture and rapid urease testing. The accuracy of the stool test was also compared with the currently accepted best noninvasive test, the carbon-13 urea breath test. Five hundred and one treatment naive patients (276 men, mean age 52 yr) were tested after endoscopy (491 patients had evaluable results). The sensitivity and specificity of the stool test were 94.1% (the 95% confidence interval was 90.6 96.6%) and 91.8% (87.3-95.1%), respectively. Pretreatment sensitivity and specificity for breath testing were similar (95.3% [92.2-97.5%] and 97.7% [94.8 99.3%], respectively). One hundred and seven infected patients were reassessed 4 wk after undergoing therapy for H. pylori infection. The posttreatment sensitivity and specificity of the HpSA assay was also excellent, although the confidence intervals were significantly wider because of the smaller number of patients (90% [68.3-98.9%], and 95.3% [88.5-98.7%], respectively). Posttreatment sensitivity and specificity for urea breath testing were also similar (90% [68.3 98.8%] and 98.9% [93.8-100%], respectively). Intercenter variability of test results did not reach statistical significance with either testing method. The authors concluded that HpSA stool testing is a reliable and easy-to-use method for diagnosing H. pylori infection in both treatment naive and posttreatment patients that compares well with carbon-13 urea breath testing. They site specific advantages of the HpSA assay to include: 1) a more simple sampling method (only one stool specimen is required); 2) the lack of a requirement for trained personnel at the testing site; and 3) there is no need for expensive equipment. PMID- 10685768 TI - The diagnosis of malign biliary obstruction with cholangiography-guided brush cytology. PMID- 10685769 TI - Mycophenolate mofetil, Crohn's disease, measles? PMID- 10685770 TI - Hyperdynamic circulation in liver cirrhosis: no evidence for peripheral vasodilation detected by ultrasound of the brachial artery. PMID- 10685771 TI - Effect of cisapride on gastroesophageal reflux disease of children. PMID- 10685772 TI - Asymptomatic retroperitoneal air after endoscopic sphincterotomy. PMID- 10685773 TI - Which scope and which technique will enhance the success rate in ERCP for the Billroth II patient? PMID- 10685774 TI - Rectus sheath hematoma clinically masquerading as sigmoid diverticulitis. PMID- 10685775 TI - Breast cancer developed in a male patient with liver cirrhosis bearing hepatocellular carcinoma. PMID- 10685776 TI - Troglitazone-associated hepatic failure. PMID- 10685777 TI - Endoscopic ultrasound and the community hospital. PMID- 10685778 TI - Fatal hepatitis associated with ranitidine. PMID- 10685779 TI - Minocycline and fulminant hepatic failure necessitating liver transplantation. PMID- 10685780 TI - Prevention of hemorrhage from intradiverticular ulcer in the duodenum by Helicobacter pylori eradication. PMID- 10685781 TI - Primary hepatic lymphoma and primary biliary cirrhosis. PMID- 10685782 TI - Garlic oil and Helicobacter pylori infection. PMID- 10685783 TI - Unsuccessful treatment results in survival of less virulent genotypes of Helicobacter pylori in Colombian patients. PMID- 10685784 TI - Parietal cell hyperplasia with deep cystic dilations: a lesion closely mimicking fundic gland polyps. PMID- 10685785 TI - Do I need an ANA? Some thoughts about man's best friend and the transmissibility of lupus. PMID- 10685786 TI - Does occupational lifting cause hip osteoarthritis? PMID- 10685787 TI - Yet another linkage reported--what does it mean? PMID- 10685788 TI - Abnormal fatty acid pattern in rheumatoid arthritis. A rationale for treatment with marine and botanical lipids. AB - OBJECTIVE: To assess the fatty acid pattern in plasma and synovial fluid (SF) in rheumatoid arthritis (RA) and to determine clinical factors related to possible abnormalities. METHODS: Thirty-nine patients with RA were included. SF samples were obtained from 9 patients. Disease activity was assessed using the Ritchie Articular Index and erythrocyte sedimentation rate. Fatty acids were assayed with gas liquid chromatography. RESULT: Decreased levels of eicosapentaenoic acid (p < 0.0001) and total n3 polyunsaturated fatty acids (p < 0.05) were observed in plasma and in joint fluid, respectively. An increase of the substrates of delta-5 desaturase (C20:3n6 and C20:2n6) and decrease of their products (C20:4n6 and C22:4n6) was observed in plasma total lipids and phospholipids. The long chain mono-unsaturated fatty acids (C20: 1n9, C22: 1n9, C24: ln9) were increased in the joint fluid and in plasma phospholipids. Patients with active disease showed a mild decrease of several saturated fatty acids, n3, and n6 polyunsaturated fatty acids. Minor abnormalities or no changes in fatty acid profile were found related to use of steroids, nonsteroidal antiinflammatory drugs, and gold salts, or malnutrition. CONCLUSION: The fatty acid pattern found in RA (decreased levels of n3 polyunsaturated fatty acids) may explain the beneficial effect of fish oil. Changes in n6 polyunsaturated fatty acids suggest that delta-5 desaturation is decreased and this might facilitate the antiinflammatory effect of botanical lipids in RA. PMID- 10685789 TI - In vivo blockade of tumor necrosis factor-alpha in patients with rheumatoid arthritis: longterm effects after repeated infusion of chimeric monoclonal antibody cA2. AB - OBJECTIVE: To investigate the longterm consequences of tumor necrosis factor alpha (TNF-alpha) blockade in patients with rheumatoid arthritis (RA), to compare changes after repeated infusion of cA2 monoclonal antibody with those occurring after the initial treatment, and to investigate significant correlations of cellular or serological changes to the duration of clinical benefit for each patient. METHODS: A clinical trial testing TNF-alpha monoclonal antibody cA2 in treatment of RA showed this therapeutic agent is highly effective. A dosage of 1 mg/kg or 10 mg/kg cA2, given in a single infusion, was compared to placebo. After clinical relapse all patients were (re)treated with 3 or 10 mg/kg cA2. In parallel to this clinical study, we investigated cellular and molecular changes induced by in vivo blockade of TNF-alpha. RESULTS: After an initial transient increase, T lymphocyte counts were not significantly different from starting values throughout the observation period. Monocyte counts as well as serum interleukin 6 (IL-6) and soluble intercellular adhesion molecule 1 (sICAM-1) concentrations remained decreased for several weeks after infusion. After a repeated infusion, increases in numbers of T cells and decreases in monocytes and IL-6 and sICAM-1 concentrations were evident again. Changes in cell counts, however, were smaller, especially in the group initially treated with the low dose (1 mg/kg), despite a higher retreatment dosage of 3 or 10 mg/kg cA2. Similarly, in this group decrease of IL-6 and sICAM-1 concentrations was less pronounced, was delayed to Day 7 after infusion, and lasted for a shorter period than seen after initial treatment. CONCLUSION: We conclude that in vivo TNF-alpha blockade leads to prolonged cellular and serological changes. This effect appears to be less pronounced after repeated infusion of cA2 compared to the initial treatment, mainly in the low dose group. PMID- 10685790 TI - A single cell analysis of Fas ligand positive T cells in rheumatoid synovial fluid. AB - OBJECTIVE: To investigate the function of Fas ligand (Fas-L) positive T cells in rheumatoid synovium, we analyzed the T cell receptor (TCR) CDR3 region and examined the expression of cytokines in both Fas-L+ and Fas-L- single T cells. METHODS: Synovial fluid (SF) samples were collected from 2 patients with rheumatoid arthritis. TCR BV8+ T cells were sorted into a 96 well plate at a density of one cell per well. Expression of Fas-L, interferon-gamma, interleukin 2 (IL-2), IL-4, IL-6, and IL-10 was analyzed by reverse transcription-polymerase chain reaction and Southern blot and the TCR BV8 junctional region was sequenced. RESULTS: Twenty-two of 30 TCR BV8+ T cells from Patient 1 and 20 of 43 TCR BV8+ T cells from Patient 2 were Fas-L+ T cells, while the others were Fas-L-. Junctional sequence analysis showed the presence of some conserved amino acid motifs in the CDR3 region (SRQ, GFG, SSG, SGS, LGTSGTL, TLSS) in 13 clones of Fas L+ T cells from Patient 1, whereas no conserved amino acid motif in Fas-L-T cells was found. In Patient 2, conserved amino acid motifs (PGQ, GQG, TTWGA) in the CDR3 region were found in 6 clones of Fas-L+ T cells, while only one was found in 2 clones of Fas-L-cells. In Fas-L+ T cells, 90-93% expressed both IL-2 and IL-10 mRNA. CONCLUSION: Fas-L+ TCR BV8+ T cells revealed the conserved amino acids motif in the CDR3 region, suggesting that Fas-L+ T cells might expand by antigen stimulation and play a crucial role as Th0-type T cells in triggering autoimmunity in rheumatoid synovium. PMID- 10685791 TI - Association between IgM response to IgG damaged by glyoxidation and disease activity in rheumatoid arthritis. AB - OBJECTIVE: To determine the association of serum IgG advanced glycation endproducts (AGE) and IgM anti-IgG-AGE antibodies with clinical measurements of rheumatoid arthritis (RA) disease activity. METHODS: The study group consisted of 62 patients with RA and 16 control patients with osteoarthritis. Patient derived variables included perceived disease activity (10 cm visual analog scale, VAS) and Health Assessment Questionnaire (HAQ) results. Clinical measures of RA activity consisted of tender and swollen joint counts and a physician evaluation of disease activity (by VAS) as well as history of nodules, bone erosions, Sjogren's syndrome, and vasculitis documented by chart review. Patient sera were evaluated for glucose, glycosylated hemoglobin, and presence of RF, IgG-AGE and IgM anti-IgG-AGE. The nitroblue tetrazolium colorimetric and aminophenyl boronic acid methods were used for measurement of IgG-AGE, along with an ELISA for measurement of IgM anti-IgG-AGE. RESULTS: Significant correlations were found between the presence of IgM anti-IgG-AGE and clinical measurements of swollen joint count and physician VAS. CONCLUSION: IgM anti-IgG-AGE appears to be associated with clinical measurements of RA activity and represents a new marker of more active disease in RA. PMID- 10685792 TI - Divergent effect of cyclosporine on Th1/Th2 type cytokines in patients with severe, refractory rheumatoid arthritis. AB - OBJECTIVE: To investigate the effect of cyclosporine on cytokine production, especially on T helper 1 (Th1) and T helper 2 (Th2) type cytokines, in patients with rheumatoid arthritis (RA). METHODS: A 16 week randomized, double blind, placebo controlled study of cyclosporine (2.5 to 4 mg/kg/day) was conducted in 40 patients with severe, refractory RA who had residual inflammation and disability despite partial responses to prior maximal tolerated dose of methotrexate (MTX; < 15 mg/week) and low dose prednisone (< 10 mg/day). Clinical and laboratory variables, and circulating levels of interleukin 2 (IL-2), IL-4, IL-10, IL-12, tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) measured by ELISA were compared between patients (cyclosporine group) treated with cyclosporine plus MTX and those (placebo group) treated with placebo plus MTX at entry and at 16 weeks. RESULTS: At 16 weeks, the cyclosporine group (n = 17), compared with the placebo group (n = 17), had greater decreases in tender joints, swollen joints, patient global assessment, patient self-assessed disability, and C-reactive protein, as well as having more patients with > 20% improvement. Comparison of circulating cytokines at entry and at 16 weeks showed significant decreases of IL-2 (median -61 vs 7 pg/ml; p = 0.004) ("+" denotes increase, "-" denotes decrease), IL-12 (median -313 vs -14 pg/ml; p = 0.002), TNF alpha (median -55 vs 5 pg/ml; p < 0.001), and IFN-gamma (median -21 vs 5 pg/ml; p = 0.003), and a significant increase of IL-10 (median 55 vs -12 pg/ml; p < 0.001) in the cyclosporine group compared with the placebo group. The degree of IL-10 increases correlated strongly with the degree of IL-12 decreases in the cyclosporine group (r = 0.572, p = 0.016). However, there was no change in circulating IL-4 between the 2 groups. Within the cyclosporine group, the improved patients (n = 10) compared to the non-improved patients (n = 7) had a greater increase in circulating IL-10 (median 172.0 vs 85.2%; p = 0.01). The rate of increase of IL-10 strongly correlated with the rate of improvement of joint scores (r = 0.718, p = 0.001) after administration of cyclosporine. CONCLUSION: Our results suggest that the therapeutic effect of cyclosporine is achieved by correcting a Th1/Th2 imbalance (a shift of Th1 type to Th2 type), which may be involved in the pathogenesis of RA; and that circulating IL-10 is useful to assess the clinical improvements in patients with RA after administration of cyclosporine. PMID- 10685793 TI - Rheumatoid synovial fluid contains bioactive leukemia inhibitory factor with cartilage degrading activity--another target for chondroprotective intervention. AB - OBJECTIVE: To determine if the procatabolic activity of inflammatory synovial fluids (SF) from patients with rheumatoid arthritis (RA) could be attenuated by the cytokine antagonists murine leukemia inhibitory factor (LIF) binding protein (mLBP) and interleukin 1 receptor antagonist (IL-1ra). METHODS: Pig articular cartilage explants were cultured in the presence of either 20% v/v rheumatoid (RA) or osteoarthritic (OA) SF and varying concentrations of either mLBP and/or IL-1ra. The catabolic activity of the SF and the relative effects of mLBP and/or IL-1ra were assessed by determining the percentage release of sulfated glycosaminoglycans from cartilage explants. LIF concentrations were measured by ELISA. RESULTS: RA SF but not OA SF stimulated release of proteoglycans from pig cartilage explants in vitro (47.3 +/- 2.2% vs 24.6 +/- 2.0%; p < 0.0001). Murine LBP at 100 ng/ml and recombinant human (rh) IL-1ra at 5000 ng/ml produced a dose dependent inhibition of this proteoglycan release (p < 0.0067 and p < 0.0111, respectively). The RA SF stimulated proteoglycan release was attenuated by mLBP and rhIL-1ra independently. No additive effect of this attenuation was observed when maximal inhibitory doses were used in combination. The decrease in proteoglycan release produced by mLBP correlated significantly with LIF concentrations in RA SF. CONCLUSION: These findings are consistent with the concept that IL-1 stimulates cartilage proteoglycan resorption in RA. They also support the hypothesis that LIF, too, contributes to cartilage proteoglycan resorption in RA. The residual stimulation not accounted for by IL-1 or LIF suggests other cytokines may contribute. The role of LIF and related or unrelated cytokines may need to be taken into account to optimize chondroprotection in RA and other rheumatic diseases. PMID- 10685794 TI - Cells expressing dendritic cell markers are present in the rheumatoid nodule. AB - OBJECTIVE: To determine if dendritic antigen-presenting cells (DC) are present in rheumatoid nodules, as has been reported in the synovial lesions of rheumatoid arthritis. METHODS: Nodules (n = 14) were examined with monoclonal antibodies (Mab) recognizing the DC differentiation/activation markers CD83, CMRF44, and CMRF56 and an antibody recognizing the CD1a antigen present on epithelial tissue associated DC. Results. Cells expressing CMRF44 were common in rheumatoid nodules, comprising 22% of nucleated cells versus 13% in synovial membranes (n = 10). Cells positive for CD1a (5%) and CD83 (2%) were less common. A majority (86%) of CMRF44 positive cells were also positive for the macrophage marker CD14. This left a significant minority of putative DC that were single stained with CMRF44. CONCLUSION: Cells bearing DC markers are as frequent in the rheumatoid nodule as in the synovial lesions. A majority are "indeterminate" cells that are CD14 positive but a proportion are single stained putative DC. The lack of lymphoid collections containing DC and T and B lymphocytes in the nodule suggests that local presentation of antigen may not occur in the rheumatoid nodule, as is thought to be the case in synovial membranes containing lymphoid follicles. This difference could potentially be explained by different states of activation, and differentiation of DC within the 2 lesions. PMID- 10685795 TI - Incidence of glenohumeral joint involvement in seropositive rheumatoid arthritis. A 15 year endpoint study. AB - OBJECTIVE: To evaluate the incidence of involvement and nature of destruction of glenohumeral (GH) joints in a prospectively followed cohort of 74 patients with seropositive and erosive rheumatoid arthritis (RA). METHODS: At the 15 year followup radiographs of 148 GH joints were evaluated, and the grade of destruction was assessed by the Larsen method. RESULTS: Erosive involvement (Larsen grade 2) was observed in 71/148 (48%) GH joints in 41/74 (55%) patients; 30 patients had bilateral and 11 unilateral involvement. The incidence of mild erosions (Larsen grade 2) was 401148 (27%), and of severe (Larsen 3-5) 31/148 (21%). The 11 most severely involved (Larsen grade 5) joints were seen in 6 (8%) patients. Erosions were most often (61/71 joints) observed on the superolateral articular surface of the humerus. Glenoidal involvement was less common (28/71 joints). The Larsen score (0-100) for peripheral joints correlated significantly with the GH joint Larsen grade on both sides (p < 0.001). CONCLUSION: After 15 years more than half the patients with RA showed definite involvement and 1 in 4 had severe destruction of the GH joint. The greatest destruction was almost always bilateral. PMID- 10685796 TI - Synovial adhesions are more important than pannus proliferation in the pathogenesis of knee joint contracture after immobilization: an experimental investigation in the rat. AB - OBJECTIVE: To measure intraarticular pannus proliferation after early and prolonged joint immobility using an animal model. METHODS: Forty rats underwent unilateral immobilization of a knee joint with an internal fixator for periods of 2, 4, 8, 16, and 32 weeks. Twenty rats received sham surgery. The knee joints were harvested and processed for histological examination. The synovial intima length and the subintimal area were measured on standardized sagittal sections with image analysis software. The measurements were recorded with regard to their location (anterior or posterior; superior or inferior). RESULTS: Intra and interrater reliabilities for all measurements were > 87.9%. The synovial intima length was smaller in immobilized knees than in controls at all time points. At 4 and 32 weeks, the difference was statistically significant (p < 0.05). The differences were marked in the posterior synovium, where the intima length of immobilized knees was significantly smaller than in controls after 4, 8, 16, and 32 weeks of immobilization (p < 0.05). The subintimal area was comparable in immobilized and control knees at all time points. CONCLUSION: We standardized the quantification of intraarticular pannus in a joint contracture model after immobility of up to 32 weeks' duration. This study revealed a significant decrease in synovial intima length but no change in the subintimal area of immobilized knees compared with controls. The decrease in synovial intima length with immobility suggests that adhesions of synovium villi rather than pannus proliferation are the major pathophysiological changes leading to contracture after immobility. PMID- 10685797 TI - Anti-MAM antibodies in rheumatic disease: evidence for a MAM-like superantigen in rheumatoid arthritis? AB - OBJECTIVE: Superantigens (SAg) are potent immunomodulatory microbial proteins that can activate T cells, B cells, natural killer cells, and monocytes and are known to trigger experimental autoimmune disease. We investigated whether sera from patients with rheumatic diseases contained elevated antibodies to Mycoplasma arthritidis mitogen (MAM) or staphylococcal enterotoxins A and B (SEA and SEB). METHODS: Standard ELISA were used to measure IgG responses to SAg and IgM and IgG rheumatoid factors and total IgM and IgG levels. Modifications of standard lymphocyte proliferation assays were used to determine functional consequences of the observed antibodies. RESULTS: Antibodies to MAM were elevated in sera from patients with rheumatoid arthritis (RA) compared to sera from patients with systemic lupus erythematosus, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, or healthy controls. Responses to other SAg were also elevated in rheumatic disease sera, but the levels were not specific for a given rheumatic disease. Anti-superantigen antibody levels did not correlate with the presence of rheumatoid factor. CONCLUSION: The selected elevation of antibodies to MAM in RA sera suggests that MAM or a MAM-like molecule might be associated with RA, whereas elevation of antibodies to SEA and SEB in sera from patients with rheumatic diseases was less specific. PMID- 10685798 TI - Determinants of health status in fibromyalgia: a comparative study with systemic lupus erythematosus. AB - OBJECTIVE: To compare perceived health status in women with fibromyalgia (FM) and systemic lupus erythematosus (SLE) using the Medical Outcomes Study (MOS) Short Form Health Survey (SF-36); and to identify determinants of physical and mental health in each patient group. METHODS: A cross sectional study of 46 women with FM (mean age 48.13 yrs, SD 9.40) and 59 women with SLE (mean age 42.36 yrs, SD 11.31). Patients with FM were recruited from a rheumatology clinic and a rheumatology practice, while patients with SLE were recruited from 4 rheumatology clinics. Clinical examination determined disease activity (by Systemic Lupus Activity Measure) in SLE and a tender point count was used for FM. Patients completed questionnaires assessing health status (SF-36), stress (Hassles), social support (Social Support Questionnaire 6), and coping (Coping Inventory for Stressful Situations). RESULTS: Patients with FM reported more impairment on the following SF-36 subscales: physical function (p < 0.001), role physical (p < 0.001), bodily pain (p < 0.001), and vitality (p < 0.001). Physical component summary scores were also significantly lower (p < 0.001) for the FM group. Four hierarchical regression analyses were computed to determine factors related to physical and mental health in each patient group, with the following variables in the equation: age, income, disease activity (Step 1), hassles (Step 2), emotional and task coping, and social support (Step 3). Better physical health in FM was related to higher income (R2 = 0.17, p < 0.05). In the SLE group, better physical health was associated with younger age, less disease activity, and lower hassles (R2 = 0.37, p < 0.0001). Worse mental health among women with FM was associated with more hassles, more emotional coping, and less satisfaction with social support (R2 = 0.64, p < 0.0001), while lower income, higher hassles, and more emotional coping were linked to worse mental health in SLE (R2 = 0.46, p < 0.0001). CONCLUSION: Health related quality of life (HRQL) is impaired among women with FM and SLE, with FM patients reporting greater impairment along several dimensions. Enhancing the HRQL of patients with FM and SLE requires targeting specific modifiable psychosocial factors. PMID- 10685799 TI - The Systemic Lupus International Collaborating Clinics/American College of Rheumatology (SLICC/ACR) Damage Index for Systemic Lupus Erythematosus International Comparison. AB - OBJECTIVE: To compare patients with systemic lupus erythematosus (SLE) from different centers with respect to demographics and Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR DI) scores, and to assess whether the SLICC/ACR DI changed over time, and whether initial DI scores were related to outcome. METHODS: Members of SLICC completed DI scores and patient demographics on patients followed in their centers. Information was provided at 2, 5-10, and > 10 years of followup. Data were entered on computer and analyzed on SPSS/PC+ and SAS using descriptive statistics and analysis of variance. RESULTS: Information for 1297 patients within 2 years of first clinic visit was submitted from 8 centers. There were 1187 women and 110 men with a mean age at diagnosis of SLE of 32 years. Seven hundred sixty-two were Caucasian, 423 were black, and the remainder were of other races. There were more blacks in the American centers than in Canadian or European centers. Five centers provided information for the 3 time periods. The DI increased over time. Ninety nine patients had died. Higher SLICC/ACR DI scores were documented in patients who went on to die. CONCLUSION: The SLICC/ACR DI is a valid measure for damage in SLE. PMID- 10685800 TI - Accurately describing changes in disease activity in Systemic Lupus Erythematosus. AB - OBJECTIVE: To determine whether Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores correlate with the clinician's impression of level of disease activity. METHODS: In total, 230 patients with SLE followed at the University of Toronto Lupus Clinic who had 5 visits 3 months apart in 1992-93 were studied. At each visit a standard protocol was completed. A clinician who did not know the patients or their SLEDAI scores evaluated each patient record and assigned a clinical activity level. "Flare" was defined by new or increased therapy for active disease, an expression of concern, or use of the term "flare" in the physician's notes. The SLEDAI score was calculated from the database. RESULTS: SLEDAI scores described a range of clinical activity as recognized by the clinician. Median SLEDAI scores ranged from 2 (inactive disease) to 8 (persistently active or flare). When the clinician assessed the patient to be improved, the median SLEDAI score decreased by 2. When the clinician assessed that the patient was experiencing a flare, the SLEDAI score increased by a median of 4. CONCLUSION: Based on our data we propose the following outcomes for patients with SLE: flare, an increase in SLEDAI > 3; improvement is a reduction in SLEDAI of > 3; persistently active disease is change in SLEDAI +/- 3; and remission a SLEDAI of 0. These outcomes will allow a more complete description of a patient's response to therapeutic intervention in a responder index. PMID- 10685801 TI - Complement degradation product C3d in urine: marker of lupus nephritis. AB - OBJECTIVE: To examine whether serum and urine C3d, a degradation product of C3, correlate with renal and extrarenal lupus activity. METHODS: Serum and urinary C3d levels were measured by ELISA in 15 healthy individuals and 24 patients with systemic lupus erythematosus (SLE) (8 with inactive disease, 7 with active but nonrenal disease, 9 with active lupus nephritis). Disease activity variables like serum C3, C4, and anti-dsDNA antibodies were also measured. RESULTS: The median serum C3d levels were significantly higher (p < 0.01) in patients with active (26 arbitrary units/ml; AU/ml) and inactive SLE (27 AU/ml) compared to healthy controls (11.25 AU/ml); levels were comparable in patients with active renal and extrarenal SLE. On the other hand, urine C3d was elevated only in patients with active SLE; its level was highest in patients with active lupus nephritis (0.87 AU/ml) compared to patients with active extrarenal diseases (0.31 AU/ml; p < 0.05), to patients with inactive lupus nephritis (0.06 AU/ml; p < 0.001), or to levels in healthy individuals (0.06; p < 0.001). Urine C3d showed stronger correlation with disease activity score (SLE Disease Activity Index) than serum C3, C4, anti-dsDNA antibodies, and serum C3d. CONCLUSION: Urine C3d is a good index of active lupus, particularly lupus nephritis. PMID- 10685802 TI - Antibodies to thrombomodulin are found in patients with lupus anticoagulant and unexplained thrombosis. AB - OBJECTIVE: To test the hypothesis that thrombomodulin (TM) may be a target for lupus anticoagulant (LAC) antibodies. METHODS: A recombinant soluble form of TM was produced and used as an antigen for an ELISA to detect antibodies to TM (TMAB). Sixty-one samples from 58 patients identified by the coagulation laboratory as having a LAC and 200 patients with unexplained thrombosis were evaluated along with 201 healthy controls. RESULTS: Eighteen (30%) of the 58 patients with a LAC and 20 (10%) of 200 patients with unexplained thrombosis had antibodies to TM. Similar antibodies were found in only 4 (2%) of 201 normal controls. TMAB show selectivity for TM lacking chondroitin sulfate, but do not otherwise have an immunodominant region. The IgG from 6 patients with TMAB was purified, and it bound TM in our ELISA. Three of the 6 IgG fractions inhibited protein C activation 40% to 70% compared to no inhibition in 7 healthy controls. CONCLUSION: Some patients with LAC and unexplained thrombosis have antibodies to TM that may arise in response to TM that has been altered and lost its chondroitin sulfate attachment. Antibodies to TM may be an important risk factor for inflammation and thrombosis in these patients. PMID- 10685803 TI - Heterogeneous behavior of anti-beta2-glycoprotein I antibodies on various commercially available enzyme immunoassay plates coated with beta2-glycoprotein I. AB - OBJECTIVE: To evaluate commercially available enzyme immunoassay (EIA) plates for the measurement of anti-beta2-glycoprotein I autoantibody (anti-beta2-GPI). METHODS: Sera from 10 patients with the antiphospholipid syndrome, and 3 monoclonal anti-beta2-GPI antibodies established from patients with antiphospholipid syndrome, were assayed for binding to solid phase beta2-GPI on 20 commercially available plates. RESULTS: Several commercially available EIA plates were found to be of potential value for the measurement of anti-beta2-GPI autoantibody. Some plates were unsuitable for anti-beta2-GPI detection, possibly due to less beta2-GPI on the plates, or to differences in the nature of conformational changes of beta2-GPI induced by the plates. CONCLUSION: Differences among EIA plates need to be considered when measuring beta2-GPI antibodies. PMID- 10685804 TI - Polymorphisms in CTLA-4 but not tumor necrosis factor-alpha or interleukin 1beta genes are associated with Wegener's granulomatosis. AB - OBJECTIVE: The genetic factors predisposing to Wegener's granulomatosis (WG) are largely unknown. T cells are clearly involved in the disease, as are the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta). The cytotoxic T lymphocyte associated antigen-4 (CTLA-4) suppresses antigen-specific immune responses by opposing the CD28 pathway, and is crucial for a balanced T cell activation. Genetic variations in the TNF-alpha, IL 1beta, and CTLA-4 genes could thus be important in WG. METHODS: Polymorphisms in the genes coding for TNF-alpha, IL-1beta, and CTLA-4 were analyzed in 32 Swedish Caucasian patients and 109 ethnically matched controls. Results. A strong association of Ctla-4 (AT)n microsatellite to WG contrasts to the negative finding of associations between TNF-alpha NcoI, IL-1beta TaqI restriction fragment length polymorphism, and WG. The prevalence of the shortest Ctla-4 allele was decreased in patients with WG compared with healthy individuals (p < 0.0001, pc < 0.0016). CONCLUSIONS: This is the first report of a T cell related gene in association with WG. The Ctla-4 itself, or a gene close to Ctla-4, may thus contribute to the pathogenesis of WG by allowing an increased T cell activation by antigen. PMID- 10685805 TI - Detection of activated complement complex C5b-9 and complement receptor C5a in skin biopsies of patients with systemic sclerosis (scleroderma). AB - OBJECTIVE: Upregulated matrix synthesis is a hallmark of systemic sclerosis (SSc). There are indications that growth factors such as platelet derived growth factor (PDGF) are involved in proliferative pathways in SSc lesions. As activated complement releases PDGF from endothelial cells, we searched for activated complement and the complement receptor for C5a (C5aR) in skin biopsies of patients with SSc. METHODS: Snap frozen sections of 8 patients with early SSc and 5 patients with longterm SSc were examined. Using monoclonal antibodies against activated complement complex C5b-9 and the C5aR, skin biopsies derived from both clinically involved and non-involved skin were examined by APAAP immunohistochemistry. RESULTS: A pattern of activated complement C5b-9 and the CSaR could be detected in SSc microvasculature. Eleven of the 13 patients (7/8 patients with early SSc) showed positive staining for C5b-9. The CSaR was detected in 6 of the 8 patients with early SSc. In 3 patients with longterm disease, C5aR expression could also be detected in non-involved skin. CONCLUSION: Activated complement and complement receptors could be detected in early and late stages of SSc skin lesions. The presence of complement receptors in non-involved skin may indicate preclinical activation of pathways resulting in growth factor dependent matrix synthesis. PMID- 10685806 TI - Urinary mercury levels in patients with autoantibodies to U3-RNP (fibrillarin). AB - OBJECTIVE: Autoantibodies to the U3 nucleolar ribonucleoprotein (RNP) fibrillarin occur in some patients with systemic sclerosis (SSc) or other connective tissue diseases and can be induced in certain mouse strains by injections of mercuric chloride, perhaps due to antigenic alteration of fibrillarin by mercury (Hg). Thus, potential occult exposure to Hg was explored in patients with SSc. METHODS: Urinary Hg levels were measured by cold vapor atomic absorption in 13 patients with antifibrillarin antibodies (11 with SSc), 39 SSc patients without antifibrillarin antibodies, and 32 healthy controls. RESULTS: Mean urinary Hg levels were significantly elevated in the antifibrillarin antibody positive patients compared to those in other patients with SSc and controls. After correction for urinary creatinine levels, mean urinary Hg levels remained significantly different than in the other 2 groups, although Hg levels in all were still within the normal or "unexposed" range. When patients and controls with low urinary creatinine levels were excluded from analysis, there were no significant differences in mean urinary Hg levels among the 3 groups. CONCLUSION: These findings suggest that further epidemiological and basic research studies of mercury are warranted in patients with SSc, especially those expressing antifibrillarin antibodies. PMID- 10685807 TI - Impaired expression of IgA Fc receptors (CD89) by blood phagocytic cells in ankylosing spondylitis. AB - OBJECTIVE: Expression of IgA Fc receptors (CD89, FcalphaR) and their occupancy by endogenous IgA were studied on blood monocytes and neutrophits to determine if FcalphaR defects could account for enhanced serum IgA and IgA-IC commonly found in patients with ankylosing spondylitis (AS). METHODS: Peripheral blood samples were obtained from 34 patients with AS, 15 patients with rheumatoid arthritis, and 34 healthy individuals. Cell surface FcalphaR was analyzed using a quantitative flow cytometry method in which blood cells were stained with anti FcalphaR monoclonal antibodies recognizing epitopes outside the IgA binding site and with F(ab')2 fragments of anti-IgA antibodies. Modulation of cell surface FcalphaR was evaluated after incubation of blood cells at 37 degrees C in absence of plasma. Biochemical characterization of iodinated FcalphaR molecules was determined by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: FcaR expression was significantly decreased on monocytes and neutrophils in patients with AS compared to control groups. FcalphaR levels were inversely correlated with serum IgA, suggesting its negative regulatory role. Modulation experiments resulted in rapid and higher FcalphaR upregulation in AS than in controls, indicating that these molecules were downregulated only at the cell surface. Moreover, analysis of the surface iodinated FcalphaR molecules by SDS-PAGE revealed higher Mr (60-90 kDa) in AS than controls (55-75 kDa), also suggesting an altered glycosylation. Analysis of receptor occupancy revealed high levels of endogenous IgA bound to monocytes and neutrophils in patients with AS, pointing to a saturation of IgA Fc receptors. CONCLUSION: We observed impaired expression of FcalphaR in patients with AS that is characterized by a downregulation process associated with post-translational alterations and enhanced binding of endogenous IgA. These alterations might lead to a defective blood clearance by FcalphaR resulting in the enhancement of IgA and IgA-IC in AS patients. Decreased FcalphaR expression represents a new marker for this disease. PMID- 10685808 TI - Rubella virus vaccine associated arthropathy in postpartum immunized women: influence of preimmunization serologic status on development of joint manifestations. AB - OBJECTIVE: To measure preimmunization rubella virus (RV)-specific IgG levels and to relate these to the development of acute and chronic (persistent or recurrent) joint manifestations following rubella vaccination. METHODS: Specific IgG was determined by whole RV enzyme immunoassays (EIA) (Abbott Rubazyme and M33, an in house method), immunoblot, neutralization domain peptide (BCH-178c) EIA, and neutralization bioassay in prevaccine samples of 268 RV seronegative women (Abbott absorbance < 0.999 units) who had received monovalent live attenuated RA27/3 strain RV vaccine in a clinical trial that recorded joint manifestations. RESULTS: Of rubella vaccinated women tested for prevaccine antibodies, 21.7% were actually positive (> or = 10 IU/ml) by M33 EIA, 33.2% had Abbott values > or = 0.250 units, and 47.6% had RV protein-specific antibody (immunoblot), while only 17.6% were positive (> or = 10 IU/ml) by neutralization domain peptide EIA and 12.7% had neutralization titers > or = 1:8. Seropositivity by the various methods was compared to recorded occurrence of acute and chronic arthropathy (arthralgia and/or arthritis) after RV vaccination. Relative to women who had no joint manifestations, prevaccine seropositivity rates for subjects with acute arthropathy were significantly (p < 0.05) lower in the Abbott test (< 0.250 units), BCH-178c peptide EIA, and neutralization bioassay, while those who also developed chronic arthropathy had significantly lower prevaccine seropositivity rates for the Abbott (< 0.250 units) and M33 EIA and neutralization bioassay. CONCLUSION: Results suggest that risk for arthropathy following RA27/3 rubella vaccination may be higher in women who have very low prevaccine levels of antibody, particularly in assays measuring functional (neutralizing) antibodies. PMID- 10685809 TI - Magnetic resonance imaging guided corticosteroid injection of the sacroiliac joints in patients with therapy resistant spondyloarthropathy: a pilot study. AB - OBJECTIVE: To evaluate magnetic resonance imaging (MRI) guided corticosteroid injections of inflamed sacroiliac (SI) joints in patients with spondyloarthropathy with therapy resistant sacroiliitis. METHODS: We performed 16 injections in 9 patients on an outpatient basis (6 men, 3 women, mean age at onset 24.7 +/- 7.5 yrs). All patients had MRI guided injection of 40 mg triamcinolone acetonide into SI joints using an open 0.2 Tesla unit. Before and 3 months after corticosteroid injection they underwent an MRI examination with a closed 1.5 Tesla unit. RESULTS: Seven of 9 patients reported subjective improvement that lasted at least a mean of 10.8 +/- 5.6 months. Subchondral bone marrow edema on fat suppressed images resolved in 8 patients after corticosteroid injection. CONCLUSION: MRI guided corticosteroid injection of SI joints appears to be an effective and safe procedure without exposure to radiation. It is a useful therapeutic modality, especially in young patients with severe isolated sacroiliitis. PMID- 10685810 TI - Epidemiology of chronic disc degeneration and osteoarthritis of the lumbar spine in Britain and Japan: a comparative study. AB - OBJECTIVE: To compare the prevalence of spinal osteoarthritis (OA) in Britain and Japan. METHODS: A total of 206 men and 188 women living in Hertfordshire, UK, and a total of 100 men and 100 women living in Miyama, Japan, aged 60-79 years were studied. Participants completed a lifestyle questionnaire, and anteroposterior and lateral radiographs of the thoracic and lumbar spine were obtained under standardized conditions. Each lumbar radiograph was graded for osteoarthritic changes according to the overall Kellgren-Lawrence (K-L) score. Gradings were also recorded separately for disc narrowing and osteophyte formation. RESULTS: British subjects were much more likely to have lumbar spine radiographs graded as K-L grade 4 severity (p = 0.05 in men, p < 0.001 in women). British men displayed a greater prevalence of disc narrowing (p = 0.08), but less severe osteophytosis (p = 0.06) than their Japanese counterparts. British women displayed more severe disc narrowing (p < 0.001) and osteophyte formation (p < 0.001). On multiple regression analysis, a higher body mass index (BMI) was associated with excess risk in the British population (OR 1.84, 95% CI 1.12-3.02), but not in the Japanese population. Differences between countries in K-L severity persisted after allowing for age and BMI, suggesting that differences in body build could not fully explain differences in lumbar spine OA in the 2 countries. CONCLUSION: We found that severe lumbar degenerative disease is more common in the UK than in a mountain village in Japan, and that differences exist in the prevalence of both osteophytosis and disc degeneration between the 2 countries. PMID- 10685811 TI - Occupational lifting is associated with hip osteoarthritis: a Japanese case control study. AB - OBJECTIVE: Hip osteoarthritis (OA) is a frequent cause of pain and disability in Western countries, but the disorder is less common in Japan. A case-control study in Britain found obesity, hip injury, and occupational lifting to be associated with hip OA among men and women. However, there are few epidemiological studies concerning factors associated with hip OA in Japan. We performed a comparable case-control study of the disorder in Japan, and contrasted the findings with those from Britain. METHODS: The study was carried out in 2 health districts in Wakayama Prefecture, Japan. Cases were men and women aged > or = 45 years listed for total hip arthroplasty due to OA over one year, and who did not have an established cause of secondary OA (e.g., rheumatoid arthritis, ankylosing spondylitis). For each case, a control was selected randomly from the general population and was individually matched to the case for age, sex, and district of residence. Cases and controls were interviewed with a structured questionnaire about medical history, physical activity, socioeconomic factors, and occupation. Measurements were made of height and weight. RESULTS: One hundred fourteen cases (103 women, 11 men) were compared with 114 controls. We found no relationship between obesity and hip OA (OR = 1.0, 95% CI 0.5-1.9; highest vs lowest thirds of distribution of body mass index). There was, however, a statistically significant association between occupational lifting and hip OA, such that regular lifting of 25 kg in the individual's first job (OR = 3.6, 95% CI 1.3-9.7) or of 50 kg in their main job (OR = 4.0, 95% CI 1.1-14.2) was associated with increased risk of hip OA. These associations remained after adjustment for potential confounding variables. In contrast, those subjects who spent > 2 h each day sitting during their first job were significantly less likely to have the disorder (crude OR = 0.5, 95% CI 0.3-0.9). This association also remained statistically significant after adjustment for potential risk factors. CONCLUSION: Our findings support the hypothesis that occupational physical activity, particularly the lifting of very heavy loads in the workplace at regular intervals, predisposes to hip OA in both Britain and Japan. The lack of association between obesity or hand involvement and hip OA in Japan suggests that the contribution of constitutional and mechanical risk factors to this disorder might differ in different populations. However, attention to manual handling in the workplace would appear an important aspect of preventive strategies against hip OA in Western and Oriental populations. PMID- 10685812 TI - Pain and hyperalgesia in osteoarthritis of the hands. AB - OBJECTIVE: Tissue damage and pain can lead to a change in the stimulus/response characteristics of the nociceptive system. Hyperalgesia has been described in experimental pain states and some clinical conditions, but has not been investigated in osteoarthritis (OA). We sought to establish the presence of hyperalgesia at the thumb in subjects with OA of the hand and to explore any relationship between sensitivity to extrinsic stimuli and the experience of clinical pain. METHODS: Subjects with OA of the hand were divided into groups according to their pain profile. The sample also included pain-free OA cases and pain-free controls. Intensity ratings were obtained for each of 3 types of pain: continuous pain, incident pain, and movement pain. Thermal and mechanical detection and pain thresholds were measured over the forearm and the carpometacarpal joint of the thumb. RESULTS: Lower thermal and mechanical pain thresholds were evident over the thumb relative to forearm in the groups with persistent pain. Subjects complaining only of incident pain, pain-free OA cases, and pain-free controls did not exhibit regional differences in sensitivity to thermal and mechanical stimuli. Increased ratings of continuous pain were associated with lower thermal and mechanical pain thresholds. Variance in movement pain ratings was predicted by mechanical forearm pain thresholds. CONCLUSION: OA in the hands is associated with cutaneous and deep hyperalgesia to thermal and mechanical stimuli. Increased levels of continuous pain are associated with more pronounced hyperalgesia. The associations of movement pain suggest the contribution of central mechanisms in the stimulus/response changes accompanying movement pain. PMID- 10685813 TI - Immunomodulatory properties of rumalon, a glycosaminoglycan peptide complex, in patients with osteoarthritis: activation of T helper cell type 2 cytokines and antigen-specific IgG4 antigen-specific igG4 antibodies. AB - OBJECTIVE: To assess the immunogenic properties of the glycosaminoglycan peptide complex Rumalon, an aqueous extract of bovine cartilage and bone marrow frequently used in patients with osteoarthritis (OA). METHODS: Sera from 31 patients with OA who had received several series of Rumalon injections (Group 1, n = 17: before therapy and after one injection series; Group 2, n = 6: after 2-3 injection series; Group 3, n = 4: after 4-8 injection series; Group 4, n = 4: after 9-18 injection series) were tested by ELISA for antibodies against Rumalon and its components as well as by a double sandwich ELISA for type 1 [interferon gamma, interleukin 2 (IL-2)] and type 2 cytokines (IL-4, IL-5, IL-10, IL-13). RESULTS: After the first injection series antibodies to Rumalon were induced in 7 of the 17 patients that were all negative before therapy. The antibodies were preferentially of the IgG4 type. IgG4 levels were increased during therapy (ELISA optical density x 1000 in Group 1: 73.9 +/- 209.5; Group 4: 1354.5 +/- 307.6), and in Group 4 all patients had developed these antibodies. Upon analysis of cytokine levels, there was a significant increase in IL-5 (Group 1 before therapy 407.4 +/- 257.1 pg/ml, Groups 3 and 4: 1409.4 +/- 963.1 pg/ml; p < 0.001) and to a lesser extent of IL-10 during therapy (Group 1 before therapy 950.2 +/- 867.8 pg/ml, Groups 3 and 4: 2817.8 +/- 3127.3 pg/ml; p < 0.05), while type 1 cytokines were not affected. CONCLUSION: Rumalon appears to have immunomodulatory properties and preferentially stimulates IgG4 antibodies via the activation of type 2 cytokines in vivo. Whether these phenomena can be correlated with the postulated therapeutic effect of Rumalon in patients with OA remains to be seen, but pain relief via release of endorphins by Th2 cells could be one explanation. PMID- 10685814 TI - Apoptotic chondrocyte death in human osteoarthritis. AB - OBJECTIVE: To define apoptotic chondrocyte death and the expression of Bcl-2, Bax, and Fas in human osteoarthritis (OA) cartilage. METHODS: Cartilage samples were obtained from patients with knee OA at the time of joint replacement surgery and from normal autopsy cases. In OA, sections were obtained both from the lesional area, usually within 1 cm of bony exposure, and from the non-lesional area, which had macroscopically normal appearance or only mild surface irregularities. Apoptosis was verified by microscopic examination of hematoxylin and eosin stained specimens, TUNEL staining, electron microscopy, and DNA ladder analysis by electrophoresis. Immunohistochemistry was done to study the expression of Bcl-2, Bax, and Fas. Apoptotic cells and Bcl-2, Bax, and Fas positive cells were counted within defined microscopic fields. Expression of Bcl 2 and Bax was verified by Western blot. RESULTS: The percentage of apoptotic cells in the lesional area was significantly higher than in the non-lesional area in cartilage from the same patient with OA, while apoptotic cells were rarely seen in normal cartilage. This result was confirmed by TUNEL stain. Many chondrocytes with chromatin condensation were verified in electron microscopy, and DNA from OA lesional cartilage revealed a DNA ladder on electrophoresis. Bcl 2 and Fas expressions were significantly higher in the OA lesional area than in the non-lesional area. On the other hand, Bcl-2 expression in normal cartilage was significantly higher than in OA cartilage. There was no significant difference in Bax expression among normal, OA lesional, and OA non-lesional cartilage. CONCLUSION: These results show that apoptotic chondrocyte death occurs more frequently in OA compared to normal cartilage and in OA lesional compared to OA non-lesional cartilage. The different expression patterns of Bcl-2 and Fas in OA lesional and non-lesional cartilage suggest that they might be involved in the pathogenesis of OA. PMID- 10685815 TI - Cell populations involved in pigmented villonodular synovitis of the knee. AB - OBJECTIVE: Pigmented villonodular synovitis (PVNS) of the knee is a tumor-like process of uncertain nature. We analyzed the involved cell populations, iron deposition, and cell proliferation in PVNS to propose a pathogenetic concept of this still elusive disease entity. METHODS: The study was performed on a series of 14 cases of localized PVNS of the knee. Histology and histochemistry were used to evaluate basic morphology and iron deposit distribution. Immunohistochemistry was performed to characterize the inflammatory cell infiltrate and to identify the proliferating cell compartments. In situ hybridization analysis using a cDNA probe against type I collagen was utilized to further characterize the mononuclear cell infiltrate. RESULTS: In addition to the classic features (mononuclear cell infiltrate, multinuclear giant cells, iron deposits, and stromal fibrosis) we observed a chronic inflammatory cell infiltrate in all PVNS samples, in which CD8 positive T cells were conspicuous. A high portion of non phagocytotic cells resorbed iron and became CD68 positive. A proportion of mononuclear cells expressed type I collagen, thus resembling B synoviocytes. CONCLUSION: Our results suggest that preexisting chronic inflammation plays an important pathogenetic role in the PVNS disease process. Chronic inflammation increases the risk of articular bleeding and probably deranges the iron processing capacity of local synovial macrophages. The resulting iron overload could lead to a shift of iron storing cells from synovial macrophages to B synoviocytes and fibroblasts. A perpetuated proliferation and activation of these cells can explain why PVNS behaves like a neoplastic process. PMID- 10685816 TI - Familial osteoarthritis and Milwaukee shoulder associated with calcium pyrophosphate and apatite crystal deposition. AB - OBJECTIVE: To characterize and define the phenotypes observed in a large Italo Argentinean kindred with osteoarthritis, chondrocalcinosis, and Milwaukee shoulder (MS). METHODS: Seventy-five members were evaluated with a history, examination, and radiographs of shoulders, spine, hands, and knees. Superior subluxation of the glenohumeral joint was graded using shoulder radiographs and tomography and nuclear magnetic resonance imaging and 3 dimensional computed tomography was performed on selected members. In 31 family members peripheral blood DNA was utilized for genetic linkage analysis of several candidate gene loci previously linked to chondrocalcinosis phenotypes, as well as those implicated in the proper patterning of skeletal elements and cartilage differentiation. In addition, direct sequence analysis of type II collagen gene (COL2A1), the gene that codes for the major structural protein of cartilage, was undertaken in 3 affected and 3 unaffected members of the family. RESULTS: MS was seen in one member of the first generation and 6 members of the 2nd generation, while 8 members of the 3rd generation showed an incomplete form of MS. Isolated superior subluxation of the shoulder was seen in 16 other family members of the 3rd and 4th generations. Osteoarthritis of the spine and peripheral joints was seen in 31 affected members, while chondrocalcinosis was observed in 6 members of the first generation. Shoulder synovial fluid from 2 patients showed the presence of both apatite and calcium pyrophosphate dihydrate crystals. Direct analysis of the COL2A1 gene indicated no known disease determining mutations in affected members, thus excluding this gene as a candidate gene in this family. Genetic linkage to several candidate loci, including the chondrocalcinosis loci on chromosomes 5p and 8q, as well as loci for HOX A and C were also excluded. Linkage analyses of other loci for the HOX B and D genes and the PAX 1 and 9 genes were uninformative in this kindred. CONCLUSION: This kindred illustrates an unusual type of osteoarthritis with secondary intraarticular and periarticular calcification and MS in the most severely affected elderly members. A search for linkage to some potential candidate genes was either excluded or uninformative. Further linkage analysis to identify potential candidate genes is in progress. PMID- 10685817 TI - Treatment of whiplash associated neck pain [corrected] with botulinum toxin-A: a pilot study. AB - OBJECTIVE: Up to 87% of patients with whiplash associated disorder (WAD) have some degree of muscle spasm that is contributory to both pain and dysfunction. Botulinum toxin A (BTX-A) produces prolonged muscle relaxation that is dose dependent and can be easily targeted to affected muscles. BTX-A therapy may be an effective form of therapy offering an alternative or adjunct to conventional modalities. We investigated BTX-A as therapy in patients with WAD. METHODS: This randomized, double blind, placebo controlled study compares outcome measures in 26 patients with chronic neck pain (WAD-II chronic) subsequent to a motor vehicle accident. One-half of the patients received 100 units BTX-A, diluted in 1 ml saline, while the other half received just saline (1 ml). Five trigger points received 0.2 ml each of injectant via a 30 gauge needle. Outcome measures included total subjective neck, shoulder, and head pain based on visual analog scales; objective total range of neck motion (ROM), and the Vernon-Mior subjective function index. Followup assessments were carried out at 2 and 4 weeks post-treatment. RESULTS: Fourteen subjects receiving BTX-A and 12 receiving saline completed the study. The treatment group showed a trend toward improvement in ROM and reduction in pain at 2 weeks post-injection. At 4 weeks post-injection the treatment group was significantly improved from preinjection levels (p < 0.01). The placebo group showed no statistically significant changes at any post treatment time. The Vernon-Mior scale revealed a trend to improvement for both groups. CONCLUSION: BTX-A treatment of subjects with chronic WAD II neck pain resulted in a significant (p < 0.01) improvement in ROM and subjective pain compared to a placebo group, but only a trend to improvement in subjective functioning. PMID- 10685818 TI - Fibromyalgia in men: comparison of clinical features with women. AB - OBJECTIVE: To describe possible differences between male and female patients with fibromyalgia syndrome (FM) in their clinical manifestations. METHODS: Five hundred thirty-six consecutive patients with FM (469 women, 67 men) seen in a university rheumatology clinic and 36 healthy men without significant pain seen in the same clinic were included in the study. Data on demographic and clinical features were gathered by a standard protocol. Tender point examination was performed by the same physician. Level of significance was set at p < or = 0.01. RESULTS: Several features were significantly (p < or = 0.01) milder or less common among men than women, including number of tender points (TP), TP score, "hurt all over," fatigue, morning fatigue, and irritable bowel syndrome (IBS). The total number of symptoms was also fewer among men and approached significance (p = 0.02) by parametric test, but reached significance (p = 0.001) by nonparametric analysis. All clinical and psychological symptoms as well as TP were significantly (p < 0.01) more common or greater in male patients with FM than healthy male controls, with the exception of IBS (p = 0.03). Patient assessed global severity of illness, Health Assessment Questionnaire disability score, and pain severity were similar in both sexes. CONCLUSION: Male patients with FM had fever symptoms and fewer TP, and less common "hurt all over," fatigue, morning fatigue, and IBS, compared with female patients. Stepwise logistic regression showed significant differences between men and women in number of TP (p < 0.001). PMID- 10685819 TI - Systemic onset juvenile idiopathic arthritis: a retrospective study of 80 consecutive patients followed for 10 years. AB - OBJECTIVE: To investigate the relationships between systemic onset juvenile idiopathic arthritis disease activity, course of the disease, and functional class according to Steinbrocker. METHODS: The records of all children with systemic onset juvenile arthritis (JA) according to the American College of Rheumatology criteria attending our center since 1971 with a minimum followup period of 3 years were reviewed. A cohort of 80 consecutive patients entered the study: 42 males, 38 females, mean age at onset 6.3 years (range 0.7-16), mean followup period 10.7 years (range 3-33). The cumulative duration of the active periods (CDAP) in months was calculated for every patient. RESULTS: Three patterns of disease course were apparent: monocyclic (subtype I), intermittent (subtype II), and persistent (subtype III). At the last control the functional class and disease activity status were evaluated. In all subtype I patients (9 cases) the disease was in remission and no patient was in class II, III, or IV. In subtype II patients (27 cases), 16 were inactive or in remission and 6 in class III. In subtype III (44 cases) 21 were inactive or in remission and 17 were in class III or IV. The equation relating the Steinbrocker class to the CDAP was calculated considering the functional outcome as the dependent variable. The linear regression equation y = 0.0083 x + 1.266 was found with a correlation coefficient r = 0.586 (p < 0.0001). The majority of our patients were treated with disease modifying antirheumatic drugs, which in many cases were effective in reducing the duration of the active phases of disease. CONCLUSION: Systemic onset JA may present with different clinical courses; the functional outcome is always good in subtype I (monocyclic), but can be poor in subtypes II and III. The severity of disability evaluated according to Steinbrocker classes is dependent on the cumulative duration of the active periods of the disease. PMID- 10685820 TI - Prominent expression of mRNA for proinflammatory cytokines in synovium in patients with juvenile rheumatoid arthritis or chronic Lyme arthritis. AB - OBJECTIVE: To examine the cytokine profiles in synovium of patients with juvenile rheumatoid arthritis (JRA) or Lyme arthritis, 2 chronic inflammatory arthritides that affect children. METHODS: We used in situ hybridization with specific riboprobes to determine the expression of mRNA for proinflammatory or antiinflammatory cytokines in synovial samples from 5 patients with early, untreated JRA, 15 patients with late, treated JRA, and 9 patients with chronic Lyme arthritis. For comparison, synovia were examined from 6 patients with rheumatoid or psoriatic arthritis, and from 9 patients with various orthopedic conditions. RESULTS: Among the children with early, untreated JRA, a median of 3 to 8% of inflammatory cells in synovial samples expressed mRNA for the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha(TNF-alpha), or interferon-gamma (IFN-gamma). Although a median of 3.9% of the cells expressed mRNA for the antiinflammatory cytokine IL-10, none had IL-4 mRNA. Most of the patients with late, treated JRA, chronic Lyme arthritis, rheumatoid, or psoriatic arthritis had mRNA for each of these proinflammatory cytokines in about 1% of the cells, whereas mRNA for the antiinflammatory cytokines was less frequent. The inflammatory cell density was much less in the synovium of patients with various orthopedic conditions, but about 1% of the infiltrating cells expressed mRNA for at least one of the proinflammatory cytokines. CONCLUSION: Patients with early or late JRA or chronic Lyme arthritis have expression of mRNA in synovial tissue primarily for proinflammatory cytokines, with less expression of antiinflammatory cytokines. PMID- 10685821 TI - Lipophagic granulomatous panniculitis with lipoatrophy mimicking arthritis with pitting edema. AB - We describe a preadolescent girl with intense ankle synovitis and pitting edema that obscured the subcutaneous origin of the inflammation. Typical nodular disease emerged after corticosteroid tapering when regional atrophy developed. PMID- 10685822 TI - A 30 year history of panniculitis. AB - The differential diagnosis of panniculitis may be challenging due to the uniform clinical presentation of different panniculitis subsets. We describe a 45-year old woman with a 30 year history of panniculitis, who had repeatedly failed to fulfill diagnostic criteria for various panniculitis subsets. Finally, erythema induratum was diagnosed and she was successfully treated with antituberculous chemotherapy. The wide spectrum of histological alterations of chronic erythema induratum as well as the sensitivity of polymerase chain reaction for Mycobacterium tuberculosis in erythema induratum lesions is discussed. PMID- 10685823 TI - Acute adrenal crisis in a patient treated with intraarticular steroid therapy. AB - Intraarticular therapy with corticosteroids can cause systemic effects such as decreased concentration of plasma cortisol, but whether this might place a patient at risk from stress induced acute adrenal failure is not known. We describe a patient who presented with lethargy, hyponatremia, and then with acute abdomen. The diagnosis of acute adrenal crisis was related to suppression of the hypothalamic-pituitary-adrenal axis by intraarticular use of corticosteroid. This was confirmed by a low basal cortisol concentration and by a short Synacthen test that elicited an increase in plasma cortisol concentration from 36 to 481 nmol/l. Within 24 h of receiving 37.5 mg of hydrocortisone, the patient rapidly improved. PMID- 10685824 TI - Sarcoidosis presenting as necrotizing sinus destruction mimicking Wegener's granulomatosis. AB - We describe a 26-year-old man who developed nasal stuffiness and palatal destruction. Biopsy of a mass in the ethmoid sinus confirmed sarcoidosis. Treatment was initiated with oral steroid and methotrexate, with marked improvement in his symptoms. Although paranasal sinus involvement in sarcoidosis is rare it should be considered in differential diagnosis of diseases causing palatal or paranasal sinus destruction. PMID- 10685825 TI - Thrombosis associated with the prothrombin G-->A20210 mutation in Behcet's disease. AB - We describe 2 cases of Behcet's disease (BD) and thrombosis who were heterozygous for the prothrombin G-->A20210 mutation. In one case, progressive uncontrolled thromboses led to death. Case-control studies are needed to support the hypothesis of the role of the prothrombin A20210 allele as a risk factor for venous thrombosis in some patients with BD. PMID- 10685826 TI - The Dunlop-Dottridge Lecture: The pathogenesis of giant cell arteritis. PMID- 10685827 TI - A review of controlled clinical trials examining the effects of antimalarial compounds and gold compounds on radiographic progression in rheumatoid arthritis. AB - At least 8 randomized controlled clinical trials have examined the effects of chloroquine or hydroxychloroquine on radiographic progression in rheumatoid arthritis (RA). At least 12 randomized controlled trials have examined the effects of either intramuscular or oral gold on radiographic progression. A review of these studies shows that hydroxychloroquine and chloroquine have minimal, if any, inhibitory effects on radiographically documented progression of bone erosions and joint destruction when used to treat RA. Intramuscular gold (with most of the data from studies of sodium aurothiomalate) appears to be better than placebo, about equal to intramuscular methotrexate (MTX), but probably not as effective as cyclophosphamide or azathioprine in its effects on radiographic progression. Auranofin appears to be better than placebo, comparable to or perhaps moderately less effective than intramuscular gold, comparable to lower dose oral MTX (7.5 mg/week), and not as effective as higher dose oral MTX (7.5-15 mg/wk) in inhibiting radiographic progression in RA. The inhibitory effects of gold compounds on proinflammatory cytokine synthesis (especially interleukin 1) offer a plausible mechanism for their inhibitory effects on bone erosion and joint destruction in RA. PMID- 10685828 TI - Radiographic assessment of disease progression in rheumatoid arthritis patients treated with methotrexate or minocycline. AB - Radiographic studies of methotrexate (MTX) treated and minocycline treated patients with rheumatoid arthritis (RA) are reviewed. A formal metaanalysis of publications of RA treated with MTX was undertaken at the time when MTX was used for patients with established RA. Thus the conclusions of that metaanalysis may not be applicable to patients treated with MTX earlier in the course of their disease. On the other hand, there are no sufficient data to conduct a formal metaanalysis of patients with RA treated with minocycline. PMID- 10685829 TI - Alternative methods for analysis of radiographic damage in a randomized, double blind, parallel group clinical trial comparing hydroxychloroquine and sulfasalazine. AB - Radiographic data of a 48-week double-blind, randomized, parallel trial comparing hydroxychloroquine (HCQ) and sulfasalazine (SASP) and its open label extension to 3 years of follow-up are analyzed in various ways. The focus of the paper is the methodological issues involved in analyses of radiographic data in a trial. Both the traditional method of increase in erosions and total score calculated on a group level, as well as individual progression on a patient level are presented. Regardless of which method is used, there is a statistically significant reduction in radiographic progression in the SASP group compared to the HCQ group. PMID- 10685830 TI - On variability of effects for a metaanalysis of rheumatoid arthritis radiographic progression. AB - Variance of an outcome that is the topic of a metaanalysis may be estimated in a variety of ways. Variances are also not used in the same way in every metaanalysis. The metaanalysis may be of a single measure expressed in its original units, or of several such measures simultaneously, or it may be done using unitless effect sizes. We present some observations and practical recommendations on the estimation and use of variances relating to these several approaches to metaanalysis, as they may apply in a metaanalysis of radiographic progression in rheumatoid arthritis. PMID- 10685831 TI - Issues involved in a metaanalysis of rheumatoid arthritis radiographic progression. Analysis issues. AB - Missing data in controlled clinical trials may create uncertainty in the results of a study based on non-missing data. We used 4 approaches of sensitivity analysis to address this problem. Radiographic progression data were used from a randomized controlled trial of patients with rheumatoid arthritis treated with leflunomide, methotrexate, or placebo for 12 months as an example. The mean change from baseline of the Sharp total radiographic score was the primary efficacy variable for the evaluation of leflunomide in comparison with placebo in the retardation of radiographic progression. Computer simulations were used in some of these approaches. The proportion of missing radiographic data was 26.4%. Result from the non-missing data showed that leflunomide was highly statistically significantly better than placebo in the retardation of radiographic progression. Results from the sensitivity analysis showed that radiographic data are sufficiently robust that it is unlikely that the missing data would have changed the conclusions from the analysis based on non-missing data. The potential effect of missing data in the results of a clinical trial may be addressed by various methods of sensitivity analysis. Computer simulation can be a useful tool in some of these approaches. PMID- 10685832 TI - Evidence from rheumatoid arthritis trials for approval: what does it mean? With special reference to using radiographic endpoints. AB - Since medicine remains largely empirical, clinical knowledge about therapy is derived primarily from experiments designed to control confounders that use inferential techniques applied to the null hypothesis model. On a public health scale, health agencies need to assess the evidentiary weight supplied for new therapies to make approval decisions. In the field of rheumatoid arthritis (RA), there are additional analytic challenges such as designation of endpoints or missing data. The intellectual architecture that underpins these exercises continues to evolve, and recently the US Food and Drug Administration released an RA Guidance Document to describe some aspects of these exercises. This article focuses on the use of measurements of radiographic endpoints in particular as an element in the evidentiary portrayal of therapeutic efficacy in RA. PMID- 10685833 TI - Influenza may influence rheumatoid arthritis. PMID- 10685834 TI - Interleukin 6 reduces serum urate concentrations. PMID- 10685835 TI - Childhood polyarteritis nodosa presenting as a relapsing movement disorder. PMID- 10685836 TI - Cyclosporine as a treatment for multicentric reticulohistiocytosis. PMID- 10685837 TI - Androgen levels of patients with ankylosing spondylitis. PMID- 10685838 TI - Ultrastructure studies of preadult Proteocephalus longicollis (Cestoda, Proteocephalidea): transmission electron microscopy of scolex sensory receptors. AB - The ultrastructure of five types of presumed sensory receptors in the scolex of preadults of Proteocephalus longicollis is described. Two types of nonciliate sensory receptors are situated on the inner surface of the lateral sucker. They differ from each other in the shape, presence, or absence of a large rootlet, electron-dense collars, desmosomes, microtubules, and/or vesicles. In addition, three types of ciliate sensory receptors are found along the edges of the lateral suckers. They can be differentiated by the length of the cilium, by the number of electron-dense collars (one or two), and by types of vesicles. Four types of vesicles were found inside the ciliate sensory receptors. One type of ciliate sensory receptor occurring in preadults differs markedly from any of the sensory receptors previously described in adult P. longicollis. PMID- 10685839 TI - Changes in Trypanosoma cruzi phospholipid turnover induced by parasite contact with cell membranes. AB - To investigate the possibility that cell contact could initiate a series of signals in both the host cell and the flagellate protozoan Trypanosoma cruzi, we studied [32P]-phospholipid turnover during parasite interaction with cellular membranes in vitro. Lipid alterations were produced in the parasite during the initial period of contact with the plasma membranes of human erythrocytes. In the presence of calcium an increment in phosphatidylethanolamine was observed with a concomitant decrease in phosphatidic acid fractions, whereas these modifications were not observed in the absence of calcium. There was an evident decrease in phosphatidylcholine and a shift in the phosphatidylinositol/lysophosphatidylethanolamine fraction among the phospholipids of major turnover in the absence or presence of calcium. Among the minor labeled species, lysophosphatidylcholine reached levels that duplicated control values, whereas the amounts of lysophosphatidylinositol, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate diminished by over 50%. All of these variations indicate that the parasite's contact with plasma membranes induces changes involving T. cruzi phospholipids and suggest the participation of these compounds in the activation of intracellular mechanisms that might be important during the life cycle of this parasite. PMID- 10685840 TI - Immunomodulatory effects in Litomosoides sigmodontis-infected Mastomys coucha. AB - The levels of parasite-specific IgG1 and IgG2 antibodies and mitogen-induced and parasite-specific proliferative T-cell responses were determined in Litomosoides sigmodontis-infected Mastomys coucha throughout an observation period of 400 days post infection (p.i.). These were compared with the respective reactions in animals that had been immunized with intrauterine stages/microfilariae of the parasite and in animals that had been challenged after immunization as determined at up to 60 days after challenge. IgG1 antibodies to adult antigen developed early and reached a plateau at 120 days p.i., whereas IgG2 antibodies were not found before day 60 p.i., increased with rising parasitemia, reached a plateau at 200 days p.i., and, in some animals, even became the predominant IgG subclass. Proliferative responses of spleen lymphocytes to concanavalin A (Con A) and lipopolysaccharides (LPS), but not Con-A-induced interleukin 2 (IL-2) production, were found to be suppressed in infected animals during patency as compared with uninfected controls. Spleen cells of infected animals showed a weak proliferative reaction to male antigen but were unresponsive to female and microfilarial antigen during prepatency and early patency. Subsequently, when microfilaremia decreased (200 days p.i.), continuously increasing responses to all antigens were observed. Immunized M. coucha developed specific IgG1 and IgG2 antibodies, and their spleen cells showed strong proliferative responses to the three L. sigmodontis antigens. Challenge infections down-regulated the proliferative responses of spleen cells to filarial antigens as early as during the prepatent phase of the challenge infection but supported existing IgG1 and IgG2 responses. PMID- 10685841 TI - A trypanosomatid protein specifically interacts with a mammalian iron-responsive element. AB - Intracellular iron homeostasis of vertebrates and invertebrates is mediated through the interaction of iron-regulatory proteins (IRPs) with mRNAs containing a bulged hairpin-loop structure termed the iron-responsive element (IRE). We detected a protein within extracts prepared from Leishmania tarentolae that specifically interacts with a mammalian IRE; mutations to the IRE that inhibit the interaction with the mammalian protein have a corresponding effect on the interaction with the L. tarentolae protein. The disassociation constant noted for the interaction of the mammalian IRE with the L. tarentolae protein was 0.7+/-0.3 microM, whereas that recorded for the interaction with the mammalian IRP under these conditions was 5+/-2 nM. The interacting L. tarentolae protein potentially places the RNA-binding site of the IRP near the root of the eukaryotic evolutionary tree. However, unlike that of the mammalian IRPs, the L. tarentolae IRE-binding activity was not induced by growth in iron-depleted media. PMID- 10685842 TI - Antigenicity of Trichomonas vaginalis heat-shock proteins in human infections. AB - Patients infected with Trichomonas vaginalis mount humoral and cellular immune responses that often do not protect against reinfection. The oxidative stressors produced by leukocytes may trigger a heat-shock-like response in T. vaginalis trophozoites, helping the parasite to survive host immune defenses. The antigenicity of T. vaginalis heat-shock proteins (HSPs) was examined by immunoprecipitation of T. vaginalis heat-induced proteins with sera from infected patients and controls. When T. vaginalis was heat-shocked, HSPs of 169-167 and 140-137 kDa were specifically recognized by sera from infected male and female patients. However, the majority of T. vaginalis HSPs were also immunoprecipitated by control sera; all sera recognized 72- to 71-kDa, 47- to 45-kDa, 38- to 37-kDa, 35-kDa, and 31-kDa heat-induced proteins. At least 15 proteins from non-heat shocked T. vaginalis were immunoprecipitated by sera from infected patients and controls, indicating that natural or cross-reacting antibodies could participate in host responses to trichomoniasis. Molecules of 158, 135, 89, and 74-72 kDa were immunoprecipitated from some non-heat-shocked parasites only by patients' sera. A 38-kDa T. vaginalis protein was immunoprecipitated only by sera from infected females and may be specific for infection in women. PMID- 10685843 TI - Up-regulation of tumor necrosis factor-alpha and interferon-gamma expression in the spleen and lungs of mice infected with the human Babesia isolate WA1. AB - We analyzed cytokine expression in mice infected with the intraerythrocytic parasites Babesia microti and WA1. In C3H/HeN mice, WA1 infections were fatal, whereas B. microti infections were resolved. We propose that the proinflammatory cytokines tumor necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma) contribute to the WA1-associated disease. WA1 infection was characterized by up regulation of TNFalpha and IFNgamma mRNA in the spleen. Previous studies in WA1 infected mice showed that pathologic lesions occurred primarily in the lungs, including pulmonary edema and intravascular margination of leukocytes. Analysis of cytokine expression in the lungs is important for an understanding of the disease process in WA1-infected mice. Expression of both TNFalpha and IFNgamma mRNA was increased in the lungs of WA1-infected mice. Immunohistochemical staining confirmed the upregulation of these proinflammatory cytokines in the lungs. Expression of TNFalpha and IFNgamma was not up-regulated in the lungs of B. microti-infected mice. The results implicate TNFalpha and IFNgamma in the pathogenesis of WA1-associated disease. PMID- 10685844 TI - Ultrastructure of the surface structures and haptor of Empleurosoma pyriforme (Ancyrocephalinae; Monopisthocotylea: Monogenea) from the gills of the teleost fish Therapon jarbua. AB - Infections with Empleurosoma pyriforme occur between successive secondary gill lamellae on both sides of the primary lamella of Therapon jarbua. The haptoral disc bears two pairs of anchors and a pair of connecting transverse bars. The attachment of the parasite to the host gill causes inflammation, erosion and degeneration of the gill epithelia. The ventral anchors consist of an inner core of irregularly arranged, electron-dense fibrils and a smooth outer core of electron-lucent fibrils, whereas the surface of the dorsal anchors is ridged. Both the dorsal and the ventral anchors may be extended or withdrawn. The connecting transverse bars consist of longitudinally arranged fibrils in an electron-dense matrix, whereas the tendons consist of fibrils, supported in a less electron-dense matrix, which interconnect the anchor erector-protractor muscles and the haptor muscles. Two types of perikarya are present. The less common type contain large multivesicular bodies and small electron-dense granules and are located only in the haptor region. The second and more common perikarya are present throughout the body surface. The cytoplasmic syncytium contains numerous electron-dense granules and electron-lucent vesicles. Beneath the syncytium, unicellular epidermal gland cells contain electron-dense granules. Neurones containing numerous electron-dense vesicles are present in the haptor region. Uniciliate presumed sensory receptors are distributed over the body surface. Groups of ciliated sensory structures are present in the forebody. Ciliated and non-ciliated presumed sensory receptors are present in the sleeve cavity of the anchors, on the haptor and in the vicinity of the oral apertures. PMID- 10685845 TI - Isolation and analysis of a new developmentally regulated gene from amastigotes of Leishmania mexicana mexicana. AB - Leishmania differentiates from the promastigote to the amastigote stage during its digenetic life cycle. Characterization of the developmentally regulated genes during that process would help to elucidate the mechanisms of gene regulation. In this study, specific fragments of mRNAs from the amastigote stage of L. mexicana mexicana were discriminated from those of the promastigote and metacyclic stages by differential display. This technique combined with spliced-leader polymerase chain reaction allowed isolation of the complete gene VG7A5. The sequence of this gene did not align with any published L. mexicana sequence. More than one copy of this gene was identified in the genome by Southern-blot analysis and was transcribed exclusively in the amastigote stage. At 20 bp upstream from the splice AG site it has a trans-splicing polypyrimidine tract. The gene encodes the subcellular localization motifs 5'-GGACT and AAGCT-3' in the 3' untranslated region of the mRNA. The open reading frame of the gene VG7A5 predicts a polypeptide of 587 amino acid residues that has a KGRR amidation motif near its carboxyl terminus, suggesting that in the mammalian host this protein may be involved in the process of acute inflammation. PMID- 10685846 TI - Inhibitory activity of anti-interleukin-4 and anti-interleukin-10 antibodies on Toxoplasma gondii proliferation in mouse peritoneal macrophages cocultured with splenocytes from infected mice. AB - Cocultures of splenocytes from Toxoplasma gondii-immunized mice or from naive mice, separated by a transwell membrane from naive macrophage layers, induced a decrease in T. gondii proliferation in macrophages in comparison with cultures without splenocytes or cocultures with splenocytes from infected mice. Interleukin 4 (IL-4) and IL-10 levels increased in cocultures of splenocytes from infected mice with naive macrophages. In contrast, the levels of these cytokines decreased in cocultures with splenocytes from immunized mice. No correlation was found between the release of interferon-gamma (IFN-gamma) and the inhibition of parasite multiplication. Cocultures with splenocytes from immunized mice induced an increase in tumor necrosis factor-alpha (TNF-alpha) levels. In contrast, in cocultures with splenocytes from infected mice, TNF-alpha production decreased. In cocultures with splenocytes from infected mice, T. gondii proliferation in macrophages was neutralized by anti-IL4 or anti-IL10 antibodies and was associated with increased TNF-alpha production. Moreover, this study demonstrates the significant combined effect of IL-4 and IL-10 on the down-regulation of macrophage-effector functions. A soluble positive signal was given by splenocytes to induce the production of TNF-alpha by macrophages. This signal was inhibited by IL4 and IL10. This process is biologically relevant in the regulation of T. gondii proliferation. PMID- 10685847 TI - First record of Cycloposthium edentatum Strelkow, 1928 from the black-striped wallaby, Macropus dorsalis. AB - The cycloposthiid ciliate Cycloposthium edentatum Strelkow 1928 is redescribed from the stomach content of the black-striped wallaby, Macropus dorsalis (Gray) following protargol staining and light and electron microscopy. This is the first record of a cycloposthiid ciliate occurring in a marsupial, as all previous records have been restricted to eutherian herbivores, especially equids. C. edentatum was found in 7 (39%) of 18 black-striped wallabies examined, but not in any of 263 animals belonging to another 20 macropodid species examined. C. edentatum is distinguished by the form of the macronucleus, the presence of a longitudinal pellicular groove, a broad posterior tail and pellicular patterning. C. edentatum specimens recovered from M. dorsalis were smaller than those reported from horses, being up to 50% shorter and 42% thinner. PMID- 10685848 TI - Helminth fauna of the wolf (Canis lupus Linnaeus, 1758) in Belorussian Polesie. PMID- 10685849 TI - Gerbils (Meriones unguiculatus) are highly susceptible to oral infection with Neospora caninum oocysts. AB - Laboratory-reared gerbils (Meriones unguiculatus) were found to be highly susceptible to oral infection with Neospora caninum (NC-Liv strain) oocysts. Gerbils fed approximately 1000 oocysts became sick or died at 6-13 days post feeding of oocysts (PFO). N. caninum was isolated in cell culture and from gamma interferon-knockout mice inoculated with homogenates of mesenteric lymph nodes of gerbils examined as early as 1 day PFO. Numerous N. caninum tachyzoites were found in ulcerative lesions in the intestines of gerbils examined at 7-9 days PFO. In a gerbil fed 10 oocysts, N. caninum tachyzoites were found in lesions in the brain. Gerbils fed 10 oocysts developed antibodies to N. caninum by 18 days PFO as determined by the Neospora agglutination test (titers > or =1:500). All gerbils remained negative for antibodies to Toxoplasma gondii as determined by the Toxoplasma agglutination test. PMID- 10685850 TI - Neospora caninum: is it really different from Hammondia heydorni or is it a strain of Toxoplasma gondii? An opinion. AB - The published data concerning Toxoplasma gondii, Hammondia hammondi, H. heydorni and Neospora caninum on one side and between T. gondii on the other were neglected by most authors. As conclusion we are convinced that there are only two valid species: Isospora (Toxoplasma) gondii and Hammondia heydorni. The first includes as a strain H. hammondi and the latter N. caninum. In any case there is absolutely no reason (with respect to general Zoological nomenclature) to create new genera! PMID- 10685851 TI - Motor and cognitive functions in newly diagnosed adult seizure patients before antiepileptic medication. AB - OBJECTIVE: Motor and cognitive functions in patients with partial or generalized onset of seizures were evaluated prior to the administration of antiepileptic medication. MATERIAL AND METHODS: Motor function, attention and memory of 52 consecutive newly diagnosed adult patients with partial or generalized seizures were assessed with neuropsychological tests. RESULTS: Patients with partial onset of seizures did not differ from patients with generalized seizures in tests of motor function or attention, nor in tests of learning and memory. Compared to controls patients with epilepsy performed significantly worse on visual motor tasks, mental flexibility and in delayed visual memory. Within the patient group as a whole lower education, higher age and symptomatic epilepsy with more abnormal CT scan findings tended to associate with worse performance in tests of concentration and mental flexibility and tests of memory. CONCLUSION: These findings indicate that newly diagnosed adult patients with partial or generalized onset of seizures prior to treatment with antiepileptic medication experience some problems in visual motor tasks, mental flexibility and memory even without the numerous risk factors for cognitive deficits in epilepsy. In newly diagnosed patients with epilepsy as a whole symptomatic etiology was associated with somewhat more pronounced cognitive problems. PMID- 10685852 TI - Weight gain following unilateral pallidotomy in Parkinson's disease. AB - OBJECTIVE: To determine the clinical correlates and infer pathogenesis of weight gain following pallidotomy in patients with Parkinson's disease (PD). BACKGROUND: Surgical ablation of the globus pallidus internus (GPi) improves levodopa induced dyskinesias, moderately improves most other "cardinal" manifestations of PD, and has been noted to result in increased weight. METHODS: We incorporated Unified Parkinson's Disease Rating Scales (UPDRS) subscales, the Beck depression inventory and feeding questionnaire data into a linear regression model in order to determine which post-surgical change(s) may lead to weight gain over the first year following pallidotomy, n = 60. RESULTS: The mean weight gain 1 year after pallidotomy was 4.0 +/- 4.1 kg. Improvement in "off" motor scores (P < 0.005), especially gait subscores (P<0.0001), and to a lesser extent improvement in "on" motor scores (P<0.05) predicted weight gain. Changes in dyskinesia ratings, mood, food intake, dysphagia, levodopa dose, weight loss in the year prior to pallidotomy, age, and duration of PD did not correlate with subsequent weight gain. CONCLUSION: The high correlation between post-pallidotomy weight gain and "off" motor scores, suggests that this phenomenon is related to some change in underlying homeostasis associated with changes in the cardinal manifestations of PD itself, rather than secondary changes resultant from the surgery. PMID- 10685853 TI - Cardiovascular dysfunction in multiple sclerosis. AB - Cardiovascular dysfunction (CD) in multiple sclerosis (MS) is related to involvement of reflex pathways in the brainstem. The battery of CD tests was applied to a group of 40 healthy subjects and 40 patients with MS, divided in 2 subgroups according to the expanded disability status scale (EDSS). The tests included: 1) postural blood pressure changes, 2) postural heart rate changes, 3) heart rate changes on inspiration/forced expiration and 4) ECG R-R interval measurement on the Valsalva maneuver. Both groups were subjected to the functional independence scale (FIM). Imaging studies were reviewed and autonomic dysfunction at other levels was explored. The results showed a statistically significant difference (P < 0.05) in all tests when comparing patients to controls. Tests 1 and 4 had the highest significance, with findings of more severe involvement in patients with a higher EDSS and lower FIM. A correlation was also found between CD and brainstem lesions on MRI (P < 0.01). A significant number of MS patients had evidence of CD. Test 1 may be considered a simple marker, in daily clinical practice, to detect subclinical CD. Subclinical CD is a cause of disability in this group of patients. PMID- 10685854 TI - Transferrin in patients with multiple sclerosis: a comparison among various subgroups of multiple sclerosis patients. AB - OBJECTIVES: To compare cerebrospinal fluid (CSF) and serum transferrin (Tf) concentrations, transferrin quotient and index in various subgroups of MS patients. MATERIAL AND METHODS: CSF and serum transferrin concentrations, transferrin quotient QTf (i.e. CSF transferrin/serum transferrin x 10(3)) and index (QTf/Qalbumin) were determined in a group of 51 patients with clinically definite or probable multiple sclerosis (MS). Patients were subdivided according to the disease form (relapsing-remitting = RR, secondary progressive = SP, primary progressive = PP; patients with RR form were further subdivided into those in the attack and those in remission), disease severity (EDSS 0-5.5, EDSS 6.0-10.0), its treatment (non-treated - including patients treated with vitamins and/ or vasodilators only, treated - i.e. glucocorticoids and/or immunosuppressants and/or (exceptionally) beta-interferon), disease duration (0-2 years, >2-10 years, > 10 years) and sex. Correlation of transferrin values with age was also performed. RESULTS: Serum transferrin was somewhat lower and significantly more frequently subnormal in PP patients in comparison with the SP form and the RR form in remission. Transferrin index was significantly higher in the PP form than in the RR as well as the SP form. Transferrin quotient was significantly more frequently subnormal in patients in remission compared to those in the attack of the RR disease. CSF transferrin as well as transferrin quotient were more frequently subnormal in patients with short disease duration (0-2 years) than in patients with longer disease duration; these parameters, however, correlated also significantly with age. CSF transferrin and transferrin quotient were higher in male than in female patients. CONCLUSION: The authors conclude that evaluation of transferrin in MS patients - along with albumin - may help to differentiate among various MS subgroups, since there are significant differences among RR, SP and PP forms. For this purpose, however, other CSF protein fractions should be evaluated in parallel in order to obtain more complex information and to establish a panel of examinations enabling multiple statistical analyses. Transferrin evaluation in MS may also be of significant theoretical interest, since transferrin is known to be involved in the regulation of iron metabolism and it may have a protective role against the oxidative stress. Moreover, transferrin is a growth factor important for proliferation of activated T lymphocytes. By means of the use of transferrin quotient and especially transferrin index, it may be possible to estimate the proportion of intra-CNS-synthesized transferrin and/or rate of specific transferrin transport across the blood-CSF barrier. Further studies are, however, needed for such an evaluation. PMID- 10685855 TI - IL-1ra serum levels in disease stages of MS--a marker for progression? AB - Interleukin-1 (IL-1) is one of the major proinflammatory cytokines expressed consistently in multiple sclerosis (MS) lesions. Interleukin-1 receptor antagonist (IL-1ra) is the only known naturally occurring specific antagonistic cytokine counteracting IL-1. Thus IL-1ra may have a downregulating potential in the disease course of MS. We analysed if circulating IL-1ra could be associated with different disease stages of MS in sera of 84 MS patients and 18 controls. IL 1ra showed considerable variations in MS patients and controls. Nevertheless we found significantly elevated serum levels in active as well as in stable disease stages compared to controls. IL-1ra levels were higher in progressive disease courses compared to relapsing-remitting MS, but not statistically significant (median: 516 versus 434 pg/ml). Further analysis with larger groups of patients and longitudinal studies will clarify if IL-1ra is useful as a prognostic serum marker in MS. PMID- 10685856 TI - Cerebellar deficit and respiratory impairment: a strong association in multiple sclerosis? AB - The aim of the study was to analyse pulmonary function and to identify reliable prognostic factors associated with respiratory abnormalities in a consecutive series of patients with multiple sclerosis (MS). Pulmonary function was evaluated by means of a battery of measures, including maximal voluntary ventilation, forced vital capacity, forced expiratory volume, in 71 consecutive patients with primary and secondary progressive MS. Respiratory impairment was common in MS patients, occurring in 63.4% of all patients, ranging from 82.9% in non ambulatory patients (with EDSS score >6.5) to 35.7% in ambulatory patients (with EDSS score <6). Severity of illness and cerebellar and mental impairment were significantly negatively associated with basal pulmonary function. Coordination plays an important role in determining respiratory abnormalities: respiratory abnormalities were found in 27 out of 32 patients (84.4%) with severe cerebellar impairment. The presence of severe cerebellar signs was associated with a very high risk of occurrence of respiratory impairment (O.R. = 6.24; 95% C.I. 1.71 22.82). Other significant variables were severity of illness (EDSS score > 6.5) (O.R. =4.71; 95% C.I. 1.42-15.66) and long disease duration (> 15 years) (O.R. = 3.39; 95% C.I. 1.01-11.42). PMID- 10685857 TI - Epidemiological surveillance of Guillain-Barre syndrome in Sweden, 1996-1997. Network members of the Swedish GBS Epidemiology Study Group. AB - OBJECTIVES: To set up an epidemiological surveillance system of Guillain-Barre syndrome (GBS) in Sweden and to evaluate the pilot practice of the system for 2 years. MATERIAL AND METHODS: A network of neurologists reported incident patients with a GBS diagnosis among a general population of 4.5 million inhabitants in Sweden during 1996 1997. Historical GBS data from the national hospital in patient registry were used for predicting incidences and controlling under reporting. RESULTS: One hundred and seventeen cases were reported. No alarm signals were identified during the study period. A re-analysis of data in 1996 revealed: 1) a higher than expected incidence in the population aged below 40 years in January, and 2) 27% under-reporting mainly associated to one hospital and to GBS patients hospitalized outside neurological departments. Threshold values for GBS incidences among the general and selected populations were obtained from corrected data. CONCLUSION: - The Swedish Network for GBS Surveillance can provide immediate support for epidemiological evaluation of suspected outbreaks or increased GBS incidence. PMID- 10685858 TI - Evoked potentials in multiple system atrophy (MSA). AB - OBJECTIVES: To study the involvement of pyramidal tracts and sensory pathways in multiple system atrophy (MSA). MATERIALS AND METHODS: Evoked potential studies were performed in 45 MSA patients suffering from either MSA of cerebellar type (MSA-C) or MSA of parkinsonian type (MSA-P). RESULTS: Motor evoked potentials were normal in all MSA patients, whereas visual and somatosensory evoked potential abnormalities were found in about 40% of the MSA patients with no significant difference between the cerebellar (MSA-C) and parkinsonian (MSA-P) subgroup. Abnormal latencies of wave III in brainstem auditory evoked potentials were significantly more frequent in MSA-C. CONCLUSIONS: Abnormalities of somatosensory, visual and auditory evoked potentials are frequent findings in MSA, whereas abnormal motor evoked potentials are not a characteristic feature of the disease. PMID- 10685859 TI - Cerebrospinal fluid C3 and C4 indexes in immunological disorders of the central nervous system. AB - OBJECTIVES: Validation of cerebrospinal fluid (CSF) indexes as a measure for intrathecal C3 and C4 production. Examination of their role in differential diagnosis of immunological disorders of the central nervous system (CNS). MATERIAL AND METHODS: Correlative study in controls (low back pain without disk herniation) between the CSF/serum ratio (Q) for albumin, and Q C3 and Q C4. Comparative study of C3 and C4 indexes in patients with CNS dysfunction due to relapsing-remitting (RR) multiple sclerosis (MS), secondary progressive (SP) MS, systemic lupus erythematosus (SLE), and human immunodeficiency virus (HIV) infection. RESULTS: Strong and statistically highly significant correlations between Q albumin and Q C3 (r=0.89, P=0.0001), and Q C4 (r=0.68, P= 0.0001). In MS patients decreased mean values for serum (RR, SP) and CSF (RR) C3, and increased C3 index mean value (RR, SP). In CNS SLE increase of mean C3 and C4 index values. In CNS HIV increase of mean C3 and C4 index values, and CSF C3 and C4 concentrations. Most individual index values were within the reference range. CONCLUSION: CSF index is a valid tool to detect intrathecal C3 or C4 production. C3 or C4 index contributes little to the differential diagnosis of immunological CNS disorders. C3 might play a pathogenic role in various immunological CNS disorders. PMID- 10685860 TI - Distribution of cerebral microembolism in the anterior and middle cerebral arteries. AB - BACKGROUND AND PURPOSE: Cerebral infarcts occur more frequently along the middle (MCA) than the anterior cerebral artery (ACA) territory. The reason(s) for this difference remains speculative. The objective of this study was to investigate the distribution of cerebral microemboli as detected by transcranial Doppler ultrasound (TCD) along the MCA and ACA territories. METHODS: Records of consecutive patients examined for the presence of cerebral microembolism during a 32-month period at the Neurovascular Laboratory were reviewed. Of the original 375 TCD studies in 268 patients, 28 studies in 24 patients demonstrated microembolic signals (MES) and monitored the MCA and ACA on the same side. TCD studies were performed on TC-2000 or TC-2020 instruments. MES positive studies were saved and off-line reviewed. MES satisfied previously established criteria. RESULTS: MES were more frequent in the MCA than the ACA in 85.7% (24/28) of studies (P < 0.01). Of the total number of MES (n = 979), 29.6% (n = 290) were detected in the ACA and 70.4% (n=689) in the MCA (P<0.01). The mean (+/- SD) intensity of MCA MES of 12.2 (+/- 2.4) dB was significantly lower than that of ACA MES of 14.8 (+/-3.2) dB (P=0.05). The mean (+/-SD) duration of MCA MES of 38.1 (+/- 45.3) ms was longer than that of ACA MES of 30.7 (+/-34.0) ms (P=0.05). CONCLUSIONS: Cerebral microembolism occurs more frequently in the MCA than the ACA, which may explain the uneven distribution of cerebral infarcts along these arterial territories. Furthermore, there are significant differences in the characteristics of ACA and MCA MES. PMID- 10685861 TI - Clinical lacunar syndromes as predictors of lacunar infarcts. A comparison of acute clinical lacunar syndromes and findings on diffusion-weighted MRI. AB - OBJECTIVES: To evaluate if patients with acute lacunar syndromes have acute lacunar infarcts or other types of cerebral lesions on diffusion-weighted MRI. METHODS: Patients with acute lacunar syndromes underwent echo-planar diffusion MRI of the brain within 3 days after stroke onset. Localization and size of lesions with hyperintense signal were determined, compared with clinical characteristics and with findings on follow-up T2-weighted MRI. RESULTS: Twenty three patients participated in the study. Thirteen patients had pure motor stroke, 1 pure sensory stroke, 8 sensorimotor stroke, and 1 ataxic hemiparesis. Twenty-two patients had at least one lesion with increased signal on diffusion weighted MR images. These acute lesions were in the internal capsule/ basal ganglia/thalamus in 13 patients, subcortical white matter in 5 patients, brainstem in 2 patients, cortex (multiple small lesions) in 1 patient, and cortex + basal ganglia in 1 patient. The median volume of the lesions was 0.6 ml on the initial examination and on follow-up, of 17 patients after 1 to 5 months, 0.5 ml. CONCLUSIONS: Almost all patients with acute ischemic lacunar syndromes have acute lesions on echo-planar diffusion-weighted MRI within 3 days after stroke onset. These lesions are mostly small and subcortical, compatible with lacunar infarcts caused by single penetrating artery occlusion, but in a minor proportion of patients (2 of 23 in our study) a cortical involvement is found. PMID- 10685862 TI - Soluble and cell surface ICAM-3 in blood and cerebrospinal fluid of patients with multiple sclerosis: influence of methylprednisolone treatment and relevance as markers for disease activity. AB - OBJECTIVE: The expression of intercellular adhesion molecule-3 (ICAM-3), a member of the Ig supergene family, is restricted to immune competent cells. Expression of soluble and cell surface ICAM-3 (s- and c-ICAM-3) is preferentially seen in the state of low activation of the immune system. We studied the relevance of the expression levels of s- and c-ICAM-3 in cerebrospinal fluid (CSF) and blood as markers for disease activity as well as the influence of high-dose methylprednisolone (MP) treatment upon the expression of s- and c-ICAM-3 in blood of patients with multiple sclerosis (MS). MATERIALS AND METHODS: A total of 33 patients (relapses n = 25, remission n = 8) with relapsing-remitting MS were included into the study. CSF and blood were acquired from all of them. Of the patients 24 were treated with high-dose MP. In those, blood was additionally collected at the 10th day of the therapy and after 3 months. Expression of c-ICAM 3 was determined by two colour FACS analysis, whereas the concentration levels of s-ICAM-3 were measured by ELISA. RESULTS: In CSF we detected a significant decrease of the expression levels of c-ICAM-3 on CD3+ T cells in 25 patients suffering from an acute relapse in contrast to 8 patients with remission (P= 0.04). In comparison to the levels before treatment and after 3 months, at the 10th day of MP treatment we obtained highly significant changes of the expression values of c-ICAM-3 both on CD3+ T cells (P = 0.0004; P= 0.005) and CD14+ monocytes/macrophages (P =0.0006; P=0.008) on the 10th day of high-dose MP treatment from 24 MS patients. CONCLUSION: The increase of ICAM-3 levels might indicate the anti-inflammatory effect of the MP treatment. It could be interesting to search for similar effects investigating the new immune modulatoring therapy forms of MS. PMID- 10685863 TI - Occult vascular malformations of the spinal cord: report of four cases not detected by angiography. AB - Angiographically occult vascular malformations (AOVMs) are reported. Although cavernous malformations, the most common type among the spinal intramedullary AOVMs, have been well documented, the natural history and clinical features of non-cavernous type AOVMs of the spinal cord remain obscure. We have evaluated four cases of non-cavernous AOVMs in the past 9 years. Although the clinical features and magnetic-resonance imaging (MRI) of three cases were similar to those previously reported, the other one had unusual features of insidious onset with limb asymmetry. The natural history of intramedullary occult AVMs remains enigmatic. One speculation for the neurological deterioration is that it results from repeated hemorrhages around the vessels or from intralumenal thrombosis. PMID- 10685864 TI - Testing the specificity of allosteric modulators of muscarinic receptors in phylogenetically closely related histamine H1-receptors. AB - Gallamine, alcuronium and W84 (hexane-1,6-bis[dimethyl-3'-phthalimidopropyl ammonium bromide]) are prototype allosteric modulators of the G-protein coupled muscarinic acetylcholine receptor family, especially of the M2-subtype. In order to probe the specificity of muscarinic allosteric modulation, we checked whether these agents interact with histamine H1-receptors which have a high homology with muscarinic receptors. Binding experiments (38 mM Na2HPO4, 12 mM KH2PO4, pH 7.5) were performed with the H1-receptor antagonist [3H]mepyramine ([3H]MEP) in guinea pig cerebellar homogenates. For the sake of comparison, binding of [3H]N methylscopolamine ([3H]NMS) at muscarinic M2-receptors was measured in porcine cardiac homogenates under identical conditions. The modulators retarded [3H]NMS dissociation (t1/2 control=1.3 min) concentration-dependently indicating their allosteric action with half-maximum effects for gallamine at EC50,discs=27 microM, for alcuronium at EC50,diss=53 nM, and for W84 at EC50,diss=170 nM. In contrast, [3H]MEP dissociation from H1-receptors (t1/2,control=2.6 min) remained unchanged up to concentrations of 1 mM of the modulators. Equilibrium binding of [3H]NMS (KD=0.46 nM, Bmax=98 fmol/mg protein) was inhibited by gallamine, elevated by alcuronium and left almost unchanged by W84, indicating negative, positive and nearly neutral cooperativity, respectively, with the radioligand. The ternary complex model of allosteric actions yielded the equilibrium dissociation constants K(A) for the binding of the allosteric modulators to free M2-receptors: K(A,gallamine)=100 nM, K(A,alcuronium)=450 nM, K(A,W84)=69 nM. In H1-receptors, more than 1,000-fold higher concentrations than in M2-receptors were required to elicit an effect on the binding of [3H]MEP (KD=1.2 nM, Bmax=205 fmol/mg protein). Half-maximal reduction was observed at 10 mM for gallamine, 1 mM for alcuronium and 92 microM for W84. In conclusion, the muscarinic modulators have little effect on the histamine H1-receptors. PMID- 10685865 TI - Caffeine inhibits a low affinity but not a high affinity mechanism for cholecystokinin-evoked Ca2+ signalling and amylase release from guinea pig pancreatic acini. AB - Caffeine has been found to inhibit the formation and action of Ca2+-mobilizing inositol 1,4,5-trisphosphate (IP3) in pancreatic acinar cells. The aim of the present study was to investigate the effects of caffeine on cytoplasmic Ca2+ concentrations ([Ca2+]i) and amylase release in response to different agonists. [Ca2+]i was determined by cytofluorometry using fura-2 as indicator and amylase release with a substrate reagent. Stimulation with low concentrations of carbachol or cholecystokinin octapeptide (CCK-8) induces [Ca2+]i oscillations whereas higher concentrations cause sustained elevation of [Ca2+]i. The less efficacious agonists pilocarpine and CCK-JMV-180 evoke oscillations only. Caffeine inhibited carbachol-induced elevation of [Ca2+]i and amylase responses in a competitive manner, abolishing the responses to low and incompletely inhibiting the responses to high concentrations of the agonist. Also, the [Ca2+]i elevations by pilocarpine were abolished by caffeine. The effects on CCK-8 induced elevation of [Ca2+]i and amylase secretion were paradoxical, the caffeine inhibition being more pronounced at high than at low concentrations of CCK-8. This enigma was further emphasized by moderate effects of caffeine on the responses to CCK-JMV-180. The results indicate that carbachol, pilocarpine and high concentrations of CCK-8 elicit IP3-mediated responses and that CCK-JMV-180 and low concentrations of CCK-8 elevate [Ca2+]i and stimulate amylase release by another signal transduction mechanism. PMID- 10685866 TI - Ca2+ entry but not Ca2+ release is necessary for desensitization of ET(A)receptors in airway apithelial cells. AB - Ca2+ transients evoked by endothelin-1 (ET-1) were measured in single cells of a human tracheal epithelial cell line using the fluorescent Ca2+ indicator fura-2. In line with a previous study, a single exposure to ET-1 (10 nM) for 10-20 s resulted in a long-lasting desensitization to a subsequent challenge by the peptide, without affecting sensitivity to agonists for other Ca2+-mobilizing receptors such as P2y or H1, respectively. In the absence of extracellular Ca2+ ET-1 elicited a Ca2+ signal of comparable amplitude as in the presence of extracellular Ca2+ but of shorter duration. Exposure to ET-1 in the absence of Ca2+ caused significantly less desensitization. Inhibition of the Ca2+ entry component of the Ca2+ transient by means of SK&F 96365, an inhibitor of Ca2+ entry, had effects comparable to Ca2+ removal. The Ca2+ transient was shortened but not significantly reduced in amplitude, and desensitization was reduced in the presence of the compound. These data demonstrate that desensitization of ET(A) receptors (ET(A)R) is promoted by transmembrane Ca2+ entry but not by Ca2+ release. PMID- 10685867 TI - Reactive oxygen species-induced aortic vasoconstriction and deterioration of functional integrity. AB - Oxygen derived free radicals and other reactive oxygen species (ROS) are involved in a variety of disease states, which can have cardiac and vascular implications. The present study was performed to investigate the mechanism of ROS-induced vasoconstriction and the influence of ROS on the functional integrity of isolated rat thoracic aorta. ROS were generated by means of electrolysis (30 mA, during 0.5, 1, 2 or 3 min) of the organ bath fluid. ROS induced a transient (approximately 60 min) vasoconstriction and the maximally induced contraction was dependent on the duration of electrolysis. Dimethyl sulfoxide (DMSO) diminished the ROS-induced vasoconstriction almost completely, indicating a major influence of hydroxyl radicals on contractility. The dual cyclooxygenase/lipoxygenase inhibitor, meclofenamate, completely prevented the ROS-induced vasoconstriction. The phospholipase A2 (PLA2) inhibitor, oleyloxyethyl phosphorylcholine, was able to reduce the vasoconstriction elicited by ROS by approximately 70%. Conversely, the specific cytoplasmic PLA2 inhibitor arachidonyl trifluoromethylketone proved ineffective in this respect. By using the specific mitogen-activated protein kinase (MAPkinase) kinase inhibitor PD98059, it was shown that the activation of extracellular-regulated kinase (ERK) MAPkinase contributes to the ROS-induced vasoconstriction. The effects of ROS on the functional integrity of the aortae were investigated, in particular with respect to receptor (alpha1-adrenoceptor) and non-receptor-mediated contractile responses (high potassium solution). In addition, both the endothelium dependent (methacholine) and endothelium independent (sodium nitroprusside) vasorelaxation were investigated before and after ROS exposure. Electrolysis periods of 0.5 and 1 min induced a modest leftward shift of the concentration response curves for the alpha1-adrenoceptor agonist methoxamine. Longer electrolysis periods of 2 and 3 min additionally decreased the maximal response to (alpha1-adrenoceptor stimulation. Methacholine induced vasorelaxation proved diminished in aortae subjected to electrolysis (0.5, 1, 2 and 3 min), whereas relaxation to sodium nitroprusside was nearly complete in all groups. KCl-induced contractions proved attenuated only after longer electrolysis periods of 2 and 3 min. This ROS-induced deterioration of functional integrity was almost completely prevented by 0.6% DMSO. From these results we may conclude that ROS induce an eicosanoid and ERK MAPkinase-mediated vasoconstriction in isolated rat thoracic aorta. In addition, exposure to ROS leads to a deterioration of functional integrity characterized by endothelial dysfunction and decreased contractile function. PMID- 10685868 TI - Pertussis toxin suppresses carbachol-evoked cardiodepression but does not modify cardiostimulation mediated through beta1- and putative beta4-adrenoceptors in mouse left atria: no evidence for beta2- and beta3-adrenoreceptor function. AB - Activation of beta1-, beta2-, beta 3- and putative beta4-adrenoceptors modifies cardiac function. These receptors are usually coupled to Gs protein, but beta2- and beta3-adrenoceptors could also couple to Gi/o proteins. The mouse heart is used increasingly for studies of genetically disrupted or overexpressed proteins, including beta-adrenoceptor subtypes. We therefore investigated in contracting mouse left atria (2 Hz, 37 degrees C) if inactivation of Gi/o proteins with pertussis toxin modifies or uncovers effects mediated through beta-adrenoceptor subtypes. The negative inotropic effects of carbachol in atria exposed to catecholamine or high calcium (6.8 mmol/l) were assumed to be mediated through activation of muscarinic receptors coupled to Gi/o. We report conditions under which incubation of left atria with 200 ng/ml pertussis toxin for 24 h nearly abolished the carbachol responses. Although it has been reported that muscarinic receptor-mediated cardiodepression has an obligatory contribution of nitric oxide, the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine (0.1-1 mmol/l) did not modify the negative inotropic effects of carbachol, inconsistent with an involvement of nitric oxide. The positive inotropic effects of (-) noradrenaline and (-)-adrenaline, mediated through beta1-adrenoceptors, were not affected by pertussis toxin. (-)-Adrenaline did not cause positive inotropic effects attributable to beta2-adrenoceptor-mediation, in the presence of CGP 20712A (300 nmol/l) to block beta1-adrenoceptors, in control atria or atria pretreated with pertussis toxin. The positive inotropic effects of (-)-CGP 12177 (1 micromol/l), a compound with agonist activity at the putative beta4 adrenoceptor, were unaffected by pertussis toxin. The beta3-adrenoceptor selective agonist BRL 37344 (1 micromol/l), in the presence of (-)-propranolol (200 nmol/l), did not cause positive or negative inotropic effects in control and pertussis toxin-treated atria. In left atria obtained from mice injected with 150 microg/kg i.p. pertussis toxin which abolished carbachol-evoked cardiode pression, the positive inotropic effects of (-)-adrenaline were antagonised by CGP 20712A. The beta2-adrenoceptor-selective antagonist ICI 118551 (50 nmol/l) did not cause additional blockade of the effects of high (-)-adrenaline concentrations in the presence of CGP 20712A, ruling out the involvement of beta2 adrenoceptors. The results with intraparenteral PTX validate our in vitro PTX method. We conclude that inhibition of murine Gi/o proteins does not alter atrial positive inotropic effects mediated through beta1- and putative beta4 adrenoceptors and does not reveal functional beta2- and beta3-adrenoceptors. PMID- 10685870 TI - K(ATP) channel blocker HMR 1883 reduces monophasic action potential shortening during coronary ischemia in anesthetised pigs. AB - ATP-sensitive potassium channels (KATP) open during myocardial ischemia. The ensuing repolarising potassium efflux shortens the action potential. Accumulation of extracellular potassium is able to partially depolarise the membrane, reducing the upstroke velocity of the action potential and thereby impairing impulse conduction. Both mechanisms are believed to be involved in the development of reentrant arrhythmias during cardiac ischemia. The sulfonylthiourea HMR 1883 (1 [[5-[2-(5-chloro-O-anisamido)ethyl]-methoxyphenyl]sulfonyl]-3-m ethylthiourea) was designed as a cardioselective KATP channel blocker for the prevention of arrhythmic sudden death in patients with ischemic heart disease. The aim of this study was to show that this compound, which has already shown antifibrillatory efficacy in dogs and rats, is able to inhibit ischemic changes of the action potential induced by coronary artery occlusion in anesthetised pigs. Action potentials were taken in situ with the technique of monophasic action potential (MAP) recording. In a control group (n=7), three consecutive occlusions of a small branch of the left circumflex coronary artery resulted in reproducible reductions in MAP duration and a decrease in upstroke velocity. In a separate group (n=7), HMR 1883 (3 mg/kg i.v.) significantly (P<0.05) reduced the ischemia induced shortening of the MAP: during the first and second control occlusion of the coronary artery in the HMR 1883-group, MAP50 duration shortened from 218.5 +/ 3.0 ms to 166.7 +/- 3.3 ms and from 219.7 +/- 4.5 ms to 164.9 +/- 1.8 ms, respectively. After HMR 1883, during the third occlusion, MAP duration decreased from 226.9 +/- 3.6 ms to 205.3 +/- 4.3 ms only corresponding to 59% inhibition. HMR 1883 also improved the upstroke velocity of the MAP, which was depressed by ischemia: in the two preceding control occlusions ischemia prolonged the time to peak of the MAP, an index for upstroke velocity, from 10.83 +/- 0.43 ms to 39.42 +/- 1.60 ms and from 12.97 +/- 0.40 ms to 37.17 +/- 2.98 ms, respectively. With HMR 1883, time to peak during ischemia rose from 12.42 +/- 0.51 ms to 25.53+/ 2.51 ms only, corresponding to an average inhibitory effect of 53.4%. The irregular repolarisation contour of the ischemic MAP was also improved. In conclusion, the present results indicate that HMR 1883 effectively blocks myocardial KATP channels during coronary ischemia in anesthetised pigs, preventing an excessive shortening of the action potential and improving excitation propagation. PMID- 10685869 TI - Effects of imidazoline binding site ligands on the growth and viability of clonal pancreatic beta-cells. AB - Pancreatic beta-cells express imidazoline binding sites which play a role in the regulation of insulin secretion, but it is not known whether ligands for these sites also affect other aspects of beta-cell physiology. In the present study, we have investigated the effects of a range of imidazoline reagents on the growth and viability of clonal pancreatic beta-cells (RINm5F and HIT-T15). Three imidazoline compounds (idazoxan, phentolamine and antazoline) were found to cause marked inhibition of beta-cell growth in a time and concentration dependent manner. Idazoxan was the most potent of these with as little as 0.5 microM causing a significant decrease in beta-cell viability (EC50 approximately 10 microM). All three imidazolines also decreased the viability of clonal beta-cells in parallel with their inhibitory effects on cell growth. These effects were not reproduced by any of a wide-range of other imidazoline compounds, including effective insulin secretagogues such as efaroxan and RX821002. The effects of the three ligands did not correlate with their relative potencies for binding to any of the well-characterised imidazoline binding sites nor to alpha2-adrenoceptors. In addition, the inhibitory responses were not antagonised by other imidazoline binding site ligands. The inhibitory effects of idazoxan on the growth of RINm5F and HIT-T15 beta-cells required as little as 3-h exposure to the imidazoline and were not readily reversible when the reagent was removed. Reductions in growth rate were accompanied by marked alterations in the morphology of the cells, which could be detected before loss of viability. Cells exposed to phentolamine showed the characteristic features of apoptosis in that the nuclei were condensed (as judged by acridine orange staining) and electrophoresis of DNA revealed the presence of oligonucleosomal fragmentation. These changes could not be detected in cells exposed to idazoxan despite the more profound reduction in viability induced by this agent. We conclude that a sub-group of imidazoline compounds can exert profoundly detrimental effects on the growth and viability of clonal beta cells but that these effects do not correlate with their binding affinity at imidazoline binding sites or alpha2-adrenoceptors. PMID- 10685871 TI - Protective effects of calcitonin gene-related peptide on guinea-pig cardiac anaphylaxis. AB - Anaphylactic events occurring in cardiac tissues can result in cardiac dysfunction via vasoconstriction and arrhythmias. Calcitonin gene-related peptide (CGRP) is the most potent vasodilator and possesses anti-arrhythmic action. We examined the influence of CGRP on cardiac anaphylaxis in guinea-pigs. In the Langendorff-perfused heart of passively sensitized guinea-pigs, antigen challenge evoked a decrease in coronary flow, left ventricular pressure and its maximum first derivatives (+/-dP/dtmax) and an increased heart rate. Antigen challenge also induced atrioventricular conduction block. Treatment with CGRP (1 or 3 nM) significantly improved the recovery of cardiac function and reduced the incidence and duration of atrioventricular block without influencing the increased heart rate. Pretreatment with capsaicin caused effects similar to those of CGRP and markedly elevated the content of CGRP in coronary effluent. Ischaemic preconditioning, induced by two cycles each of 5 min global ischaemia and 5 min reperfusion, also improved cardiac function and raised the level of CGRP in coronary effluent. The protective effects of ischaemic preconditioning were abolished in the presence of the CGRP receptor antagonist CGRP8-37. Histamine release did not differ significantly during any of the interventions. The findings of the present study indicate that, in guinea-pig hearts, CGRP protects against cardiac anaphylaxis and that the cardioprotection by CGRP is independent of histamine release. PMID- 10685872 TI - Androgen deprivation, estrogen treatment and vascular function in male rat aorta. AB - The beneficial effects of estrogen on arterial function in women are well established, whereas studies concerning the vascular role of androgens have produced conflicting results. In the present study, we examined the effects of androgen deprivation and of estrogen treatment on vascular responses in male rats. Vascular reactivity was studied in aortic rings excised from intact and castrated rats, which had been implanted with capsules containing either 17beta estradiol (E2) or its vehicle for 5 days. Contractile responses to noradrenaline were potentiated by castration and by E2 treatment. Concentration-response curves for N-methyl-L-arginine and superoxide dismutase indicated that the tone-related release of NO increased in tissues from castrated, compared with intact rats, but was not affected by E2 treatment. Endothelium-dependent relaxation elicited by carbachol and histamine were not altered by castration and were attenuated by E2 in preparations from intact, but not from castrated rats. Moreover, aortic prostacyclin release dropped by about 40% after E2 treatment in tissues from both intact and castrated animals. Similarly, smooth muscle sensitivity to NO significantly decreased following castration and E2 treatment, as assessed by responses to sodium nitroprusside. Finally, no differences among groups were detected in platelet thromboxane A2 production. Thus, vascular responses in male rats were not improved by androgen deprivation alone or by E2 treatment, whose effects differed in the presence or absence of androgens. These findings provide evidence for the gender specificity of the vascular effects of estrogen and may be consistent with a beneficial role of physiologic levels of male sex hormones in arterial function. PMID- 10685873 TI - Gender and hypertension interact to regulate neuropeptide Y receptor responsiveness. AB - We have compared the interaction between gender and strain (normotensive Wistar Kyoto vs. spontaneously hypertensive rats) in the regulation of cardiovascular and renal Y1 and Y5-receptor-mediated responses to neuropeptide Y. Anaesthetized rats received intravenous infusions of 0.3-10 microg/kg per min neuropeptide Y (45 min each) or vehicle. Enhancements of mean arterial pressure, renovascular resistance, diuresis and natriuresis were measured. Parallel group comparisons were performed on male normotensive, female normotensive, male hypertensive and female hypertensive rats. Gender and strain significantly affected the cardiovascular and renal neuropeptide Y responses. While neuropeptide Y elevated mean arterial pressure and renovascular resistance in all four groups, the extent of the alterations differed up to two- to threefold between genders and/or strains. Neuropeptide Y-induced diuresis and natriuresis were similar in male and female normotensive rats, desensitized in male but augmented in female hypertensive rats. We conclude that previously reported cardiovascular and renal neuropeptide Y responses are regulated by gender and the presence of hypertension. However, at least for renal function alterations in female hypertensive vs. male normotensive rats cannot be predicted from those in male hypertensive and female normotensive rats. PMID- 10685874 TI - Noradrenoceptor antagonism with idazoxan improves L-dopa-induced dyskinesias in MPTP monkeys. AB - Treatment of Parkinson's disease with L-dopa is plagued in a majority of patients by dyskinesias. Noradrenaline/dopamine interactions are proposed on behavioral, biochemical, physiological and anatomical grounds. The aim of the study was to test the potential antidyskinetic effect of the alpha2-adrenoceptor antagonist, idazoxan, in a primate model of Parkinson's disease. Six female cynomolgus monkeys previously rendered parkinsonian by the toxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) and presenting an unchanged syndrome for several months were used. All responded readily to L-dopa but had developed dyskinesias which were manifested with each dose. In the first part of the study, seven doses of idazoxan (ranging from 0.25 mg/kg to 10 mg/kg, p.o.) were administered together with the vehicle or in combination with a fixed dose of L-dopa/benserazide (100/25 mg, p.o.). In the second part of the study, a fixed dose of idazoxan (7.5 mg/kg) was administered daily for 10 days and L-dopa was added to idazoxan on days 1, 4, 7 and 10. Vehicle (empty capsule) was used as control. Idazoxan, by itself (ranging from 5 mg/kg to 10 mg/kg), increased locomotor activity and improved the disability score with virtually no dyskinesias in three animals. In combination with L-dopa, idazoxan did not impair the antiparkinsonian response but significantly reduced dyskinesias in all six animals up to 65% at doses of 7.5 mg/kg and 10 mg/kg and delayed their onset, so that the "ON" state without dyskinesias was prolonged. The antidyskinetic effect of idazoxan was maintained when repeatedly administered for 10 days. On day 10, the locomotor response to L dopa was significantly potentiated by chronic administration of idazoxan. Our results indicate that idazoxan has some antiparkinsonian effect of its own and may constitute a useful adjunct to L-dopa as it can reduce dyskinesias without impairing the relief of symptoms, this effect being maintained over time in this model. PMID- 10685875 TI - A comparison of the acute effects of zotepine and other antipsychotics on rat cortical dopamine release, in vivo. AB - The acute effects of systemic administration of the antipsychotic drug, zotepine, on extracellular dopamine (DA) in the frontal cortex of freely-moving rats were studied using in vivo microdialysis and compared with the actions of clozapine, olanzapine and haloperidol. Treatment with zotepine (1.0 mg/kg, i.p.) resulted in a prolonged elevation of cortical DA levels for up to 180 min post-drug. A maximal rise of +333% was observed at 120 min post-zotepine treatment. Clozapine (10.0 mg/kg, i.p.) also evoked a rise in extracellular DA which was similar in duration (200 min) to that resulting from treatment with zotepine. A maximal rise of +223% was observed at 100 min post-clozapine treatment. Olanzapine (1.0 mg/kg, i.p.) resulted in an immediate increase in DA levels which was maximal 40 min post-treatment (+280%) with levels returning to pre-injection values by 100 min after dosing. In contrast, haloperidol (0.1 mg/kg, i.p.) had no measurable influence on cortical DA levels. Local perfusion with the NA uptake inhibitor, nisoxetine (10 microM), resulted in an increase in cortical DA levels which was maximal at 100 min post-onset of perfusion (+257% above baseline). Administration of zotepine (1.0 mg/kg, i.p.) during nisoxetine perfusion elevated DA levels to a maximum of +301% above baseline, 60 min post-zotepine. These results show that acute administration of each of three drugs with an atypical antipsychotic profile causes an elevation of cortical DA in freely-moving rats at doses relevant to those derived from animal models which predict antipsychotic activity. As a dysfunction in cortical DA is thought to be involved in both the negative symptoms of schizophrenia and cognitive deficits in schizophrenic patients, it is possible that zotepine's ability to elevate cortical DA levels may underlie its effectiveness in successfully treating these components of schizophrenia. Furthermore, the ability of zotepine to elevate cortical DA is more likely to derive from its inhibition of the NA transporter rather than DA receptor blockade in this region. PMID- 10685876 TI - Effects of inescapable shock and conditioned fear on the release of excitatory and inhibitory amino acids in the locus coeruleus. AB - We investigated the importance of endogenous amino acids in the locus coeruleus in inescapable electric shock and conditioned fear. In naive rats and in rats exposed to noise (N), light (L) and electric shock (S) or to N + L only, the locus coeruleus was superfused with artificial cerebrospinal fluid through a push pull cannula and the release of GABA, taurine, glutamate, aspartate, serine and glutamine was determined in the superfusate by HPLC after derivatization with o phthaldialdehyde. Locomotor activity, arterial blood pressure and heart rate were telemetrically monitored. The placement of naive rats or conditioned rats from their home cage to a chamber provided with a grid-floor for shock virtually did not change the release rates of the amino acids in the locus coeruleus. Motility was enhanced in naive and conditioned rats to a similar extent. Blood pressure and heart rate were enhanced in conditioned rats only. Exposure to N + L + S for 5 min greatly enhanced the release rates of all determined amino acids in the locus coeruleus. In conditioned rats the increase in release of most amino acids lasted longer than in naive rats. Electric shock also enhanced motility, blood pressure and heart rate. In conditioned rats, motility and cardiovascular changes were more pronounced and/or lasted longer than in naive rats. Exposure of conditioned rats to the conditioned stimuli N + L for 5 min led to an increased release of taurine and aspartate. The enhanced release of taurine lasted 30 min. Exposure to N + L did not affect the release rates of amino acids in naive rats. N + L did not influence motility but arterial blood pressure and heart rate were elevated in conditioned rats. The findings show that inescapable electric shock enhances the release of several amino acids in the locus coeruleus, while conditioned fear selectively increases the outflow of taurine and aspartate. Moreover, conditioned fear prolongs the response of excitatory and inhibitory amino acids to electric shock. The results suggest that an excitatory amino acid (aspartate) and an inhibitory amino acid (taurine) of the locus coeruleus are implicated in conditioned fear. PMID- 10685877 TI - Gabapentin-lactam, a close analogue of the anticonvulsant gabapentin, exerts convulsant activity in amygdala kindled rats. AB - The cyclic GABA analogue gabapentin (GBP), which recently has been marketed for treatment of epilepsy, is particularly effective against complex-partial seizures as occurring in temporal lobe epilepsy. In the present study, we compared the effects of GBP and its lactam analogue (GBP-L) in the amygdala kindling model of temporal lobe epilepsy. In fully kindled rats, GBP (50 mg/kg and 100 mg/kg i.p.) dose-dependently increased the threshold for focal seizures and inhibited the progression from focal to generalized seizures. This effect was not associated with any marked adverse effects. In contrast, GBP-L (10-50 mg/kg) induced myoclonic activity and generalized clonic seizures in kindled rats, demonstrating a striking qualitative difference between the two compounds. By comparison with non-kindled rats it was shown that kindling markedly enhanced the sensitivity of rats to the convulsant activity of GBP-L. The finding that the anticonvulsant efficacy of GBP is lost by lactam formation indicates that GBP and GBP-L differ in their mechanism(s) of action. PMID- 10685878 TI - Changes in NMDA receptor subunit gene expression in the rat brain following withdrawal from forced long-term ethanol intake. AB - Changes in mRNA levels of N-methyl-D-aspartate receptor (NR) subunits were studied in a rat model of withdrawal from forced ethanol ingestion over a period of 8 days. In part, this model may reflect the epsilon-type of human alcoholism according to Jellinek (College University Press, New Haven; 1972). The epsilon type is characterized by dipsomania over a period of several days, recurring every few months and often followed by ethanol-induced seizures. Seizures may be modulated by an increased glutamatergic neurotransmission to excitatory or inhibitory neurons on the basis of a changed gene expression of NR subunits. This hypothesis promoted the present study. Film autoradiograms and emulsion-coated brain sections following labeling of cholinergic and GABAergic neuron populations were evaluated. NR subunit 1 (NR1) expression, studied with a probe recognizing all NR1 transcripts, was unchanged after withdrawal from chronic ethanol treatment compared to control animals. Using probes specific for different splice segments of NR1, however, we found that, in ethanol-treated rats, the expression of NR1-2 was decreased in all, and that of NR1-4 in all but one, areas investigated (only single label experiments were performed with NR1 splice variants). Withdrawing rats revealed a higher expression of NR subunit 2A (NR2A) mRNA in GABAergic neurons. No changes could be observed at the regional level. Conversely, NR2B mRNA was not substantially altered in cholinergic and GABAergic neurons, but showed a decrease over brain areas. For both, NR2C and NR2D, no ethanol-related changes of mRNA expression were observed. A link between such differential alterations in NR mRNA subunit expression and ethanol-induced seizures in withdrawing alcoholics of the epsilon-type seems possible. PMID- 10685879 TI - Selective inhibition of monoamine neurotransmitter transporters by synthetic local anesthetics. AB - Synthetic local anesthetics (LAs) have been found to have cocaine-like characteristics with some psychotomimetic action, possibly through monoaminergic neurotransmission. To gain insight into the relation between LA action and monoamine transporters, we investigated the effect of synthetic LAs on neurotransmitter transporters, including monoamine transporters. We used cloned transporter cDNAs and examined transient functional expression in COS cells and stable expression in HeLa cells. Among the LAs tested, procaine and other ester type LAs inhibited [3H]DA uptake and binding of [3H]2-beta-carbomethoxy-3-beta-(4 fluorophenyl)tropane (CFT), a cocaine analogue, in COS cells expressing rat dopamine transporter (DAT). The inhibition was concentration-dependent. The inhibitory effect on [3H]DA uptake was reversible and not dependent on pH, as observed in HeLa cells stably expressing DAT. Procaine also inhibited uptake of norepinephrine (NE) and serotonin (5-HT) by the norepinephrine transporter (NET) or serotonin transporter (SERT) expressed in COS cells. On the other hand, procaine and other LAs had little or no effect on [3H]GABA and [3H]glutamate uptake in COS cells expressing mouse GABA or rat glutamate/aspartate transporter. IC50 values for [3H]DA uptake inhibition correlated well with those for [3H]CFT binding inhibition, but not with intrinsic anesthetic potency. Kinetic analysis of monoamine uptake inhibition by procaine in COS cells expressing rat DAT, NET or SERT revealed a competitive action similar to that of cocaine. These results demonstrate that certain LAs selectively inhibit monoamine transporters. This might contribute to the cocaine-like psychotomimetic action of certain LAs. PMID- 10685880 TI - An innovative method for rapid characterisation of phospholipase C activity: SB242,084 competitively antagonises 5-HT2C receptor-mediated [3H]phosphatidylinositol depletion. AB - 6-chloro-5-methyl-1-[6-(2-methylpyridin-3-yloxy) pyridin-3-ylcarbamoyl] indoline (SB242,084) is a novel, selective 5-HT(2C) receptor antagonist, but its actions at these sites have been little characterised at the cellular level. We employed a rapid and innovative approach to investigate its functional activity at phospholipase C (PLC)-coupled human 5-HT(2C) receptors expressed in CHO cells. PLC activity was determined as a decrease in the [3H]phosphatidylinositol ([3H]PI) content of cell membranes. Serotonin (5-HT) stimulated [3H]PI depletion (pEC50=8.74), and SB242,084, like mesulergine, completely reversed this action of 5-HT (pK(B)=9.25 and 9.01, respectively). Further, in Schild analysis, SB242,084 behaved as a high affinity competitive antagonist, inducing a parallel, rightward displacement of the 5-HT stimulation isotherm without loss of maximum efficacy. The pA2 of 9.50 was similar to its binding affinity (pKi=9.38). SB242,084 also displayed antagonist properties when PLC activity was examined by conventional determination of [3H]inositol phosphate generation. Employing this parameter, the potency of SB242,084 (pK(B)=9.21) and that of mesulergine (pK(B)=9.06) closely resembled those determined by [3H]PI depletion. In conclusion, determination of [3H]PI depletion constitutes a useful and novel technique to characterise agonist and antagonist properties of ligands at PLC-coupled receptors. PMID- 10685881 TI - Efficacy of synthetic zeolite to reduce the toxicity of aflatoxin in broiler chicks. AB - Synthetic zeolites (NaX, NaY, NaA, and CaA) were evaluated in vitro for their ability to sorb aflatoxin (AF) B1 from an aqueous solution. Zeolite NaA (ZN) was selected to be tested in vivo because of its high affinity and its stable association with AFB1. This sorbent was incorporated into diets (1%) containing 2.5 mg/kg AFB1. Male broiler chicks from 21 to 42 d of age received ad libitum access to their respective diets and water. When compared with controls, BW gains were lower (P < 0.05) for broilers that were fed AF in their diets. No differences were found between the BW gains of chicks fed diets without AF and those of chicks fed AF + ZN, indicating almost total protection against the effects caused by AF. Liver weights were considerably higher in chicks fed a diet containing AF, compared with those of controls, nevertheless, no significant differences were found in feed:gain ratio among the groups. The findings of this research suggest that ZN can counteract some of the toxic effects of AF in growing broiler chicks. PMID- 10685882 TI - Lactobacillus flora in the cloaca and vagina of hens and its inhibitory activity against Salmonella enteritidis in vitro. AB - Lactobacilli in the cloaca and vagina of 40 normal laying hens were investigated quantitatively and qualitatively, and their ability to inhibit growth of Salmonella enteritidis (SE) was examined using a spot-the-lawn technique. All samples of cloacal contents and half the samples of vaginal mucus were positive for lactobacilli. The means +/- SD of total Lactobacillus counts in the cloaca and those in the vagina were log10 5.5 +/- 1.1 and 2.5 +/- 2.6 cfu/g, respectively. In the cloaca, Lactobacillus acidophilus was isolated from 92.5% of hens, and Lactobacillus salivarius was isolated from 85.0% of hens, whereas Lactobacillus fermentum was isolated from only one hen. In the vagina, L. acidophilus and L. salivarius were isolated from 42.5% of hens. In the inhibition assay in vitro, all strains of Lactobacillus from cloacal contents and vaginal mucus inhibited growth of SE. There was a wide range of the inhibitory activity even in the same species. No difference of the growth inhibition zone was observed between lactobacilli from cloaca and those from vagina. The present study suggested that lactobacilli in the cloaca and vagina of hens might have a protective effect against SE colonization. PMID- 10685883 TI - Efficacy of neomycin sulfate water medication on the control of mortality associated with colibacillosis in growing turkeys. AB - The objective of this investigation was to evaluate the efficacy, safety, and toxicity of neomycin sulfate (Neomix 325) water medication to control mortality associated with colibacillosis (Escherichia coli) in growing turkeys. One efficacy trial was conducted at five locations; each location included 2,880 sexed 21-d-old turkey poults that were naturally challenged with litter from turkey flocks that had colibacillosis. Between 5 and 7 d after challenge, and when mortality had reached 0.5%, poults were randomized within sex into three treatment groups of 0, 11, or 22 mg neomycin sulfate/kg body weight. In each location, each treatment was replicated 12 times with 40 poults per sex per replicate. All treatments were administered in the drinking water for 5 d. The pivotal decision criterion was mortality. Mortality was defined as 1) supported mortality (SM): positive microbial culture for E. coli and gross lesions, 2) diagnosed mortality (DM): diagnosed as associated with E. coli but not supported by lesions or positive microbiological cultures, 3) overall mortality (OM): mortality associated with E. coli or other microorganisms and miscellaneous reasons such as accidents (trampling or suffocations). Performance data (growth and feed utilization) also were measured and are reported without statistical analysis. Results from this efficacy study clearly demonstrated the effectiveness of neomycin sulfate against E. coli as measured by a reduction in mortality. In the target animal safety and toxicity study (done in conjunction with the efficacy study), neomycin sulfate in the drinking water at 66, 110, or 220 mg/kg per d for 15 d had no observable adverse effects on poult performance, as measured by feed or water consumption, body weight, gross pathology, or mortality. PMID- 10685884 TI - Effects of photoperiod and melatonin on lymphocyte activities in male broiler chickens. AB - Understanding the role of the pineal gland in regulating the immune response and the role of photoperiod in influencing pineal gland secretions are becoming increasingly important. The purposes of the present experiments were to investigate the effects of different photoperiod regimens on T- and B-lymphocyte activities in broiler chickens. Next, the influence of different photoperiod regimens on the responsiveness of lymphocytes to melatonin in vitro was examined. The effect of melatonin in vitro on lymphocyte activities was also studied, regardless of the photoperiod received. Finally, the effects of photoperiod on the profiles of different splenocyte cell types were investigated. To study the effect of photoperiod on lymphocyte activities, different photoperiod regimens were used. These were: constant lighting, 23 h light:1 h darkness; intermediate lighting, 12 h light:12 h darkness; and intermittent lighting, 1 h light:3 h darkness. Peripheral blood and splenic lymphocyte activities were tested at 3 and 6 wk of age by performing a mitogen cell-proliferation assay with a polyclonal T cell mitogen, concanavalin A (Con A), and T-dependent B-cell mitogen, pokeweed mitogen (PWM). To study the effect of photoperiod on the responsiveness of lymphocytes to melatonin in vitro or the effect of melatonin in vitro on lymphocyte activities regardless of photoperiod received, lymphocytes from the chickens that were exposed to the different photoperiod regimens were incubated with mitogen and different concentrations of melatonin. To study the effect of photoperiod on profiles of different cell types, the percentages of splenocyte subpopulations from birds exposed to different photo-periods were determined using flow cytometry with CD4+, CD8+, CD3+, and B-cell markers. The results of these studies indicate that splenic T and B lymphocytes from 6-wk-old chickens grown in intermittent lighting had higher activities than those from chickens grown in constant lighting. Peripheral blood and splenic lymphocytes from chickens raised under constant lighting were more responsive to melatonin in vitro than those from chickens raised under intermittent lighting. This difference in response may be due to lower levels of melatonin in birds receiving constant lighting, making them more sensitive to melatonin in vitro. Melatonin in vitro enhanced the mitogenic response of peripheral blood T lymphocytes from 6-wk old chickens, splenic T lymphocytes from 3-wk-old chickens, and splenic T and possibly B lymphocytes from 6-wk-old chickens. Finally, intermittent lighting increased the percentages of splenic CD4+, CD8+, and CD3+ cells but not B-cell subpopulations at 6 wk of age, presumably because of increased levels of melatonin in birds receiving intermittent lighting. Our results re-emphasize the importance of melatonin in regulating host immune response; this regulation could be accomplished through exposing broiler chicks to intermittent lighting. PMID- 10685885 TI - Effects of moniliformin on performance and immune function of broiler chicks. AB - Three trials were conducted to evaluate the effect of moniliformin (M) on performance and immune function in chicks. Day-old chicks were randomly assigned to four dietary treatments (0, 50, 75, or 100 mg M/kg diet). In Trial 1, chicks were placed on treatments for 3 wk and were injected intravenously with 4.6 x 10(6) Escherichia coli on Day 21. Blood samples were collected at 60, 120, and 180 min after inoculation, and liver, spleen, and lung were collected at 180 min postinjection. Compared with control chicks, chicks fed 75 and 100 mg M/ kg diet had higher (P < 0.05) numbers of E. coli colonies in the circulation, liver, and spleen. In Trial 2, chicks were placed on diets for 4 wk and were injected with 0.5 mL Newcastle disease virus (NDV) vaccine intramuscularly on Weeks 2 and 3 of the experiment. The primary and secondary anti-NDV antibody titers were measured 7 d after each injection. Chicks fed 100 mg M/kg diet had lower (P < 0.05) secondary antibody titers than did control chicks. In Trial 3, lymphocyte proliferation in chicks exposed to M in vivo and in vitro was determined. Results of the in vivo study showed that cell proliferation in response to mitogens from control- and M-fed chicks did not differ (P > 0.05). For the in vitro study, lymphocyte proliferation decreased linearly (P < 0.01) with increased concentrations of M. In all three trials, chicks fed 100 mg M/kg diet had lower (P < 0.05) feed intake and weight gain than did control chicks. Data from the current study suggested that M decreased performance and immune response in chicks at the level of 75 mg/kg diet. PMID- 10685886 TI - Effects of Salmonella typhimurium lipopolysaccharide on broiler chickens. AB - The effects of Salmonella typhimurium lipopolysaccharide (LPS) on the physiology of 3-wk-old broiler chickens were studied at 12, 24, and 48 h after a single intravenous injection of saline or LPS. Lipopolysaccharide elevated cloacal temperature by 3 h after injection, induced a diuretic response, and decreased BW gain. An increase in the relative liver weight was evident in LPS-treated birds at all time intervals, whereas a decrease in the relative weight of bursa of Fabricius was observed only at the 48-h time point. The plasma interleukin (IL)-6 and the blood heterophil concentrations were elevated at 12 and 24 h following LPS administration. These changes were not observed in control chickens or in LPS treated chickens at 48 h. A decrease in the blood glucose concentration in LPS treated birds at 12 h was accompanied by an elevation in the blood phosphate level. An increase in total plasma protein concentration was observed only at 24 and 48 h after LPS treatment. Comparative SDS-PAGE analysis of plasma proteins from these birds under nonreducing conditions showed some quantitative differences in four bands of proteins between saline and LPS-treated chickens. A protein corresponding to an approximate molecular weight (MW) of 65 kDa increased in LPS-treated chickens, and three other proteins with MW of approximately 39, 49, and 56 kDa showed reductions in concentration compared with saline-treated controls. These results show that LPS induces a number of physiological changes that may be responsible for the regulation of the acute phase response in chickens. PMID- 10685887 TI - Chicken intestinal aminopeptidase: partial sequence of the gene, expression and activity. AB - Aminopeptidases are members of a membrane-bound metallopeptidase family that are expressed at a high level on the brush-border membrane of enterocytes. Because the rapid growth of meat-type chickens depends on the dietary supply of amino acids, a study of intestinal aminopeptidases, which play a central role in protein digestion, is important. This study is the first reported isolation of the partial cDNA of chicken intestinal aminopeptidase and sequencing of a 1.7-kb cDNA fragment. The gene was isolated by reverse transcriptase polymerase chain reaction using six primers chosen from conserved regions of the aminopeptidase genes. Amplified fragments were extracted from the gel, purified, and sequenced. By using this chicken cDNA as a probe, northern blot analysis revealed a transcript of approximately 3.5 kb in the chicken duodenum, jejunum, and ileum tissues. Higher RNA expression and activity of aminopeptidase were found in the ileum tissue compared with the duodenum and jejunum segments. PMID- 10685888 TI - Autosomal albino chicken mutation (ca/ca) deletes hexanucleotide ( deltaGACTGG817) at a copper-binding site of the tyrosinase gene. AB - We compared tyrosinase cDNA sequences from a line of autosomal albino and Black Silky chickens isolated from cultured melanocytes by reverse transcription polymerase chain reaction (RT-PCR). Both sources produce a single DNA fragment of predicted normal tyrosinase size. Direct sequencing of the PCR product showed three mutated sites in the tyrosinase gene of the albino chicken. Two silent point mutations and a deletion of six nucleotides (-deltaGACTGG) at 817 bp in the tyrosinase cDNA sequence were observed when compared with the White Leghorn and Black Silky cDNA sequences. The deduced albino chicken tyrosinase protein lacks two amino acids, aspartic acid and tryptophan. The position of these amino acids is consistent with one of the potential copper-binding sites that should be indispensable for function of the enzyme. We speculate that the six-base deletion is responsible for the inactive tyrosinase in this line of albino chickens. PMID- 10685889 TI - Incorporation of different polyunsaturated fatty acids into eggs. AB - An experiment was carried out to examine thoroughly the relationships among different n-3 and n-6 polyunsaturated fatty acids (PUFA) in the diet, their deposition into the eggs' fat, and their effect on hens' laying performance. A diet enriched with 4% fish oil (FO) was fed to the birds throughout the 14-wk laying period (Treatment 1; T1); this was the same oil source that was replaced in proportions of 25, 50, 75, or 100% with four different fat sources, resulting in 17 isocaloric dietary treatments: linseed oil (LO; T2 to T5), rapeseed oil (RO; T6 to T9), sunflower oil (SO; T10 to T13), and tallow (T; T14 to T17). Performance parameters were recorded weekly and analyzed on the basis of the replacing fat source. At the end of the 14-wk experimental period, eggs were collected, and their fatty acid (FA) profile was determined. Performance parameters were not significantly different among grouped treatments. Smaller proportions of FO in diets resulted in lower values of saturated and higher values of n-6 FA contents, regardless of the fat source used when replacing FO. The n-6 content increased mostly because of the rise in linoleic acid (LA), although the level of arachidonic acid (AA) was always higher when FO was completely suppressed. The amount of the different n-3 long-chain PUFA was lower (P < 0.001) when FO was present in lesser proportions in the diet. However, the slope of the decline of these FA changed according to the included fat. Replacing FO with LO resulted in the lowest decline of its derivatives by elongation and desaturation and an increase in the total n-3 FA in the form of linolenic acid (LNA). PMID- 10685890 TI - The effect of dietary betaine in Eimeria acervulina-infected chicks. AB - Two experiments were conducted to evaluate the effect of dietary betaine in broiler chicks with either chronic (CHR; 2.5 x 10(5) sporulated oocysts on Day 1, 4, 7, and 10) or acute (ACT; 1.0 x 10(6) sporulated oocysts on Day 1) Eimeria acervulina infections. Three hundred (Experiment 1) or 600 (Experiment 2), 4-d old male chicks were used in the 14-d experiments. In both experiments, a 2 x 3 factorial arrangement of treatments was used: two levels of betaine (0 or 0.075%) and three levels of coccidiosis infection (uninfected, CHR, or ACT). Each treatment was replicated five (Experiment 1) or 10 (Experiment 2) times with 10 chicks per replicate. In Experiment 1, the ACT infection decreased (P < 0.01) average daily gain and gain:feed, and the CHR infection decreased (P < 0.02) average daily gain. The ACT and CHR infections decreased (P < 0.06) Day 7 plasma carotenoids and Day 14 plasma total protein, and the ACT infection also decreased (P < 0.06) Day 7 plasma total protein. Average daily gain and Day 7 plasma total protein were increased in CHR chicks fed betaine but were decreased in uninfected chicks fed betaine (CHR x betaine; P < 0.09). Chicks fed betaine had decreased (P < 0.06) Day 7 plasma carotenoids. In Experiment 2 the CHR and ACT infections decreased (P < 0.01) average daily gain, average daily feed intake, grain:feed ratio, Days 7 and 14 plasma carotenoids, and Day 7 plasma total protein. Chicks fed betaine had increased (P < 0.07) average daily gains, gain:feed ratios, and lesion scores. Day 14 plasma carotenoids and plasma total protein were decreased in uninfected chicks fed betaine but were increased in CHR chicks fed betaine (CHR x betaine; P < 0.04); plasma carotenoids also were increased in ACT chicks fed betaine (ACT x betaine; P < 0.05). Betaine did not consistently affect growth performance, plasma constituents, or lesion score in CHR or ACT coccidiosis infected chicks. PMID- 10685891 TI - Effects of phosphorolytic and cell wall-degrading enzymes on the performance of growing broilers fed wheat-based diets containing different calcium levels. AB - A study was conducted to determine the cumulative effects of phosphorolytic enzymes, cell wall-degrading enzymes, and citric acid and Ca levels on feed intake, BW gain (BWG), feed conversion, intestinal viscosity, and toe ash of broilers (d 1 to 21) fed wheat-based diets. Broilers were fed the following six diets at either 0.59, 0.69, or 0.79% Ca: 1) a negative control (NC) diet, 0.17% available P; 2) NC + 750 phytase units/kg diet; (3) phytase + 3,156 units of acid phosphatase/kg diet; 4) phytase + acid phosphatase + 1,900 units of pectinase/g diet; 5) phytase + acid phosphatase + pectinase + 3% citric acid; and (6) NC plus 0.24% available P. The 18 dietary treatments were fed to four pen replicates of eight birds each. Phytase addition at the low Ca level increased BWG, improved feed intake and conversion and toe ash, and reduced intestinal viscosity and ileal length. Subsequent addition of acid phosphatase, at 0.69% Ca, resulted in increases in BWG, 42%; feed intake 32%; feed conversion 7.5%; and toe ash, 63% over the NC diet. Pectinase addition produced further improvements in 21-d BWG and feed intake at 0.59 and 0.79% Ca, increased toe ash in chicks fed 0.79% Ca, and reduced intestinal viscosity. Supplementation of wheat-based 0.17% available P diets with phytase and acid phosphatase and with appropriate concentrations of pectinase, citric acid, and Ca significantly improved BWG, feed intake and conversion and intestinal viscosity over the 0.41% available P diets. Bone mineralization of chicks fed phytase + acid phosphatase and 0.69% Ca and those fed phytase + acid phosphatase + pectinase + citric acid and 0.59% Ca was similar to that of chicks fed the 0.41% available P diets. PMID- 10685892 TI - Research notes: The effect of different levels of palm kernel meal in layer diets. AB - Palm kernel meal (PKM), a by-product from the African Palm oil industry that is extensively cultivated in tropical countries, is an interesting feed ingredient for poultry due to its availability and low cost. The objective of this study was to evaluate the use of different levels of PKM in layer diets. This particular PKM contained 9.70% crude protein, 0.20% methionine, 0.36% lysine, and a TMEn value of 2,254 kcal/kg. A control diet based on corn and soybean meal and five different levels of PKM added to it were fed to Single Comb White Leghorn hens from 18 to 38 wk of age. The PKM levels were 0, 10, 20, 30, 40, and 50%. The hens were housed three per cage (30.5 cm wide x 45.7 cm deep). The six treatments were assigned randomly to three contiguous cages in each of eight rows in a randomized complete block design. Egg production was recorded daily, and feed consumption for an entire week was recorded every 21 d. Egg weight and specific gravity were recorded for 3 consecutive d every 21 d. Mortality was recorded daily. Results show that egg production was significantly decreased (P < 0.05) only with 50% PKM in the diet. Feed conversion was not affected by any level of PKM. Specific gravity was slightly but significantly (P < 0.05) decreased by all levels of added PKM. Feed consumption, mortality, and egg weight did not differ significantly among the treatments. We concluded that this particular PKM may be used up to 40% in the diet, taking into account that specific gravity may be slightly decreased. PMID- 10685893 TI - Soluble factors and the emergence of chick primordial germ cells in vitro. AB - Previous observations obtained from a culture of blastodermal cells on a mouse fibroblast feeder layer (STO) suggested that STO cells provide a factor or factors that facilitate development of avian primordial germ cells (PGC) from dispersed embryo cells. The purpose of the current study was to test the hypothesis that soluble factors produced by STO cells are responsible, at least in part, in supporting the development of PGC in culture and to examine the effect of stem cell factor (SCF), ciliary neurotrophic factor (CNTF), and basic fibroblast growth factor (bFGF) in the development of PGC in culture. Blastodermal cells on gelatin-coated plastic or on feeder layers of CV-1 cells yielded a small number of PGC. When blastodermal cells were cultured on STO cells, a marked increase in PGC was observed. The addition of STO cell conditioned medium (STO-CM) to blastodermal cells cultured on gelatin-coated plastic and on feeder layers of CV-1 cells resulted in a significant increase in the number of PGC, indicating that soluble factors produced by STO cells can enhance the development of chicken PGC in culture. Supplementation of blastodermal cells with SCF (100 ng/mL) or CNTF (2 ng/mL) or with CNTF and SCF together resulted in a significant increase in the number of PGC after 48 h of culture on feeder layers of CV-1 cells. However, addition of bFGF (100 ng/mL) did not increase PGC. We concluded from these observations that the culture of blastodermal cells on feeder layers of STO and CV-1 cells can be used as a useful biological system in examining the regulatory factors that govern the ontogeny of the germ cell lineage in the avian embryo. PMID- 10685894 TI - Ultrastructure of spermatozoa from Japanese quail. AB - A monoclonal antibody (mAb) to epitopes on mitochondria from turkey spermatozoa cross-reacted with Japanese quail spermatozoal mitochondria. However, the pattern of binding was different from that observed for turkey sperm. The ultrastructure of quail spermatozoa was examined to determine the reason for this difference in antibody binding pattern. Light microscopy, as well as scanning (SEM) and transmission (TEM) electron microscopy, were used to study the morphology of spermatozoa from Japanese quail. Japanese quail had a sauropsid type of sperm cell, which is typical of nonpasserine birds. The spermatozoa were vermiform in shape, with a maximum width of 0.6 microm and an overall length between 230 and 250 microm. An acrosome (3.7 to 4.5 microm), nucleus (20.8 to 23.8 microm), midpiece (160 to 170 micro/m), and tail (40 to 60 microm) were observed. The TEM showed an acrosomal cap surrounding a perforatorium that inserted into the nucleus at the posterior end. Only a distal centriole was observed, which gave rise to a central axoneme with a 9+2 microtubular structure. The axoneme was encased by a spiraled mitochondrial sheath in the midpiece region (64 to 74% of the overall length of the sperm), and mitochondria numbers were estimated to be greater than 1,400 per sperm. In contrast, turkey sperm contain short midpieces with only 20 to 30 mitochondria per sperm. Differences in binding patterns of the mAb to turkey mitochondria between quail and turkey sperm were due to the presence of mitochondria on the exceptionally long midpieces of quail sperm. PMID- 10685895 TI - Immunolocalization of progesterone and estrogen receptors in the sperm storage tubules of laying and diethylstilbestrol-injected immature hens. AB - The goal of this study was to determine whether sperm storage tubules (SST) in the uterovaginal junction (UVJ) contain receptors for progesterone and estrogen (PR and ER) and whether estrogenic stimulation induces activation of these receptors in the SST. Frozen sections of UVJ obtained from immature chicks treated with or without diethylstilbestrol (DES), which is an estrogenic substance, and those from laying hens were immunostained for PR and ER. In laying hens, immunoreactions for PR were observed in the SST cells and on the surface epithelium of UVJ; the strongest reactions appeared in the SST cells. In contrast, ER immunoreactions were localized in the SST cells but not in the other cells. Immunoreactions for PR and ER were negligible in the UVJ of control immature birds that received only the oil vehicle. However, in the DES-treated birds, PR immunoprecipitates were localized in the surface epithelium and SST cells, and there were ER in the SST cells. These results suggest that the SST cells in the UVJ contain PR and ER, and estrogenic stimulation may play a significant role in inducing activation of these receptors. PMID- 10685896 TI - Effect of meat temperature on proteins, texture, and cook loss for ground chicken breast patties. AB - Soluble proteins, myofibrillar proteins, collagen, texture, and cook loss were evaluated at different meat temperatures by heating ground and formed chicken breast meat in brass containers in a water bath to temperatures of 40, 50, 60, 70, or 80 C. The soluble proteins decreased by approximately 90% as meat temperature increased from 23 to 80 C. The myofibrillar protein subunits of molecular weight greater than 43 kDa decreased with increasing temperature from 23 to 80 C as analyzed via SDS-PAGE. Amount of soluble collagen more than doubled when meat temperature was increased from 50 to 70 C. The maximum peak shear force was obtained, via Warner-Bratzler shear test, at 60 C for ground chicken breast patties. The weight of patties decreased approximately 10.3% when meat temperatures were increased from 23 to 80 C. Overall, heating temperature affected the texture of the meat and caused changes in proteins and cook loss. PMID- 10685897 TI - Denaturation of myofibrillar proteins from chickens as affected by pH, temperature, and adenosine triphosphate concentration. AB - The susceptibility to denaturation of myofibrillar protein from chicken muscles was investigated and compared with denaturation of myofibrillar protein from pork. Immediately postmortem, the Pectoralis profundus (white muscle) and the Pubo-ishio femorale (red muscle) of six Arbor Acres chickens were collected. The Semimembranosus (white muscle) and Psoas major (red muscle) of three Yorkshire x Landrace and three Yorkshire x Landrace x Duroc pigs were collected at 45 min postmortem. Protein denaturation was prevented by keeping the muscles at 0 to 2 C in a buffer (pH 7.2) containing ethylene glycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) (to sequester Ca ions). After purification, myofibrils were incubated at 25 or 40 C, pH 5.4 or 6.5, with 0, 0.68, or 3.4 mM adenosine triphosphate (ATP). Protein solubility, an indicator of denaturation, was assessed after 0, 10, 20, and 60 min incubation. Protein solubility of chicken pectoralis myofibrils was not affected by any of the conditions. In the other myofibrils, pH 5.4 caused significantly (P < 0.05) more protein denaturation than pH 6.5, and incubation at 40 C resulted in significantly more protein denaturation than incubation at 25 C. The presence of ATP (tested at pH 6.5) affected denaturation; higher ATP concentrations resulted in increased loss of solubility. We concluded that chicken red myofibrillar proteins are equally susceptible to denaturation as are pork red and white myofibrils. Chicken pectoralis (white) muscle fibers are least susceptible to denaturation. The results of this study indicate that factors other than protein denaturation are responsible for the low water-holding capacity of pale, soft, exudative chicken breast muscle. PMID- 10685898 TI - Histological characterization of hemorrhages in muscles of broiler chickens. AB - Hemorrhages in meat of broiler chickens are major quality defects. The objective of our study was to characterize the various types of hemorrhages in thigh and breast muscles with respect to their morphological appearance, location, and origin. Chickens were stunned using a water-bath stunner and were either exsanguinated and fixed or perfused with fixative. The morphological appearance of the hemorrhages was determined by the type of tissue in which they were found and by the amount of extravasating blood. Origins of hemorrhages were found only at sites of rupture of venous structures, such as postcapillary venules and small collecting veins. The absence of significant leukocyte infiltration strongly indicated that muscle tissue damage and hemorrhage occurred within the 24 h preceding stunning and slaughter. The locations and types of hemorrhages indicate different underlying mechanisms. PMID- 10685899 TI - Thermal gelation properties of spent hen mince and surimi. AB - Thermal gelation properties of spent hen mince and surimi were investigated. The mince from 98-wk-old spent hens was washed two times with 0.1% NaCl. A portion of unwashed and washed mince was mixed with 4% sugar, 4% sorbitol, and 0.2% Na tripolyphosphate to produce surimi and was kept frozen at -20 C. The mince and surimi were ground with 3% NaCl and a small amount of water to adjust the final moisture content to 80%. A 6 to 8% potato starch was mixed with some pastes. The pastes were stuffed into sausage casings and heated by one-step and two-step heating. The effects of washing, heating, and addition of ingredients on the color, composition, and functional properties of the mince and gel were compared. Washed, spent hen mince was lighter and less red in color and higher in collagen, gel strength, water-holding ability, and cooking yield than unwashed mince. The best temperature and time schedule for the gelation of spent hen mince was 90 C for 15 min in one-step heating. Heating at 100 C for 5 min after preheating at 60 to 70 C for 30 min resulted a gel with distinctly improved gel strength. Sucrose (4%), sorbitol (4%), and Na-tripolyphosphate (0.2%) improved the gel quality of nonfrozen mince but showed little cryoprotective effect against the degradation of frozen-stored product. A 6% potato starch improved the gel texture, cooking yield, and water-holding ability compared with 8% starch. PMID- 10685900 TI - Quality and sensory characteristics of selected post-rigor, early deboned broiler breast meat tenderized using hydrodynamic shock waves. AB - Our first objective was to determine the effects of explosive amount and distance of the explosive to the meat surface in the Hydrodyne process on broiler breast tenderness. Early deboned (EB) breasts were removed immediately after initial chill (45 min postmortem), stored for 24 h (4 C), and subjected to one of four Hydrodyne treatments (200 g at 20 cm, 350 g at 23 cm, 275 g at 20 cm, or 350 g at 20 cm). Breasts were water-cooked (78 C internal). Hydrodyne treatment (HYD) of 350 g at 20 cm produced the greatest reduction (28.3%) in Warner-Bratzler shear (WBS, 1.9-cm wide strips). This combination was the only treatment to improve tenderness (peak force 4.3 kg) to a level equivalent (P > 0.05) to aged controls (CA; peak force 3.1 kg). The second objective was to determine the quality and sensory characteristics of Hydrodyne-treated (350 g explosive at 20 cm) broiler breasts as compared with CA and EB. The WBS values (1.0-cm wide and thick strips) for CA (1.56 kg) were different from both HYD (3.7 kg) and EB breasts (4.7 kg). The CA resulted in more tender, flavorful, and juicer breasts than EB and HYD. The EB was higher in initial moisture release than HYD. The EB breasts with tenderness problems can be tenderized by the Hydrodyne process based on WBS results. However, higher levels of explosive may be required to optimize the tenderness improvement of EB breasts that vary significantly in initial tenderness. PMID- 10685901 TI - Cardiac natriuretic peptides: a physiological lineage of cardioprotective hormones? AB - Vertebrate hearts from fish to mammals secrete peptide hormones with profound natriuretic, diuretic, and vasodilatory activity; however, the specific role of these cardiac natriuretic peptides (NPs) in homeostasis is unclear. NPs have been suggested to be involved in salt excretion in saltwater teleosts, whereas they are proposed to be more important in volume regulation in mammals. In this review, we consider an alternative (or perhaps complementary) function of NPs to protect the heart. This hypothesis is based on a number of observations. First, evidence for NPs, or NP-like activity has been found in all vertebrate hearts thus far examined, from osmoconforming saltwater hagfish to euryhaline freshwater and saltwater teleosts to terrestrial mammals. Thus the presence of cardiac NPs appears to be independent of environmental conditions that may variously affect salt and water balance. Second, cardiac stretch is a universal, and one of the most powerful, NP secretagogues. Furthermore, stretch-induced NP release in euryhaline teleosts appears relatively independent of ambient salinity. Third, excessive cardiac stretch that increases end-diastolic volume (EDV) can compromise the mechanical ability of the heart by decreasing actin-myosin interaction (length-tension) or through Laplace effects whereby as EDV increases, the wall tension necessary to maintain a constant pressure must also increase. Excessive cardiac stretch can be produced by factors that decrease cardiac emptying (i.e., increased arterial pressure), or by factors that increase cardiac filling (i.e., increased blood volume, increased venous tone, or decreased venous compliance). Fourth, the major physiological actions of cardiac NPs enhance cardiac emptying and decrease cardiac filling. In fish, NPs promote cardiac emptying by decreasing gill vascular resistance, thereby lowering ventral aortic pressure. In mammals a similar effect is achieved through pulmonary vasodilation. NPs also decrease cardiac filling by decreasing blood volume and increasing venous compliance, the latter producing a rapid fall in central venous pressure. Fifth, the presence of NP clearance receptors in the gill and lung (between the heart and systemic circulation) suggest that these tissues may be exposed to considerably higher NP titers than are systemic tissues. Thus, a decrease in outflow resistance immediately downstream from the heart may be the first response to increased cardiac distension. Because the physiology of cardiac NPs is basically the same in fish and mammals, we propose that the cardioprotective effects of NPs have been well preserved throughout the course of vertebrate evolution. It is also likely that the cardioprotective role of NPs was one of the most primordial homeostatic activities of these peptides in the earliest vertebrates. PMID- 10685902 TI - Fecal glucocorticoids: a noninvasive method of measuring adrenal activity in wild and captive rodents. AB - To determine the utility of fecal corticosteroid concentration as a measure of chronic stress under laboratory and field conditions, we biochemically and physiologically validated a radioimmunoassay for corticosteroids in three rodent species, house mice (Mus musculus), deer mice (Peromyscus maniculatus), and red back voles (Clethrionomys gapperi). The biochemical validations demonstrated that the assay accurately and precisely measured corticosteroid concentration in the feces. The physiological validation indicated that the assay was sensitive enough to detect the stress associated with (a) brief handling and bleeding of animals, (b) chronic caloric restriction, (c) exposure to a novel environment, and (d) exposure to a novel cold environment. Our results suggest that fecal measurements reflect stress levels experienced by these animals approximately 6-12 h before defecation. Therefore, given a judicious trapping and trap-monitoring protocol, this assay has considerable utility for measuring the stress levels at which animals actually exist in the field. PMID- 10685903 TI - The ventilatory responses of the caecilian Typhlonectes natans to hypoxia and hypercapnia. AB - Typhlonectes natans empty their lungs in a single extended exhalation and subsequently fill their lungs by using a series of 10-20 inspiratory buccal oscillations. These animals always use this breathing pattern, which effectively separates inspiratory and expiratory airflows, unlike most urodele and anuran amphibians that may use one to many buccal oscillations for lung inflation and typically mix expired and inspired gases. Aquatic hypoxia had no significant effect on the breathing pattern or mechanics in these animals. Aerial hypoxia stimulated ventilatory frequency and increased the number of inspiratory oscillations but had little effect on inspiratory and expiratory tidal volume. Aquatic hypercapnia elicited a large significant increase in air-breathing frequency and minute ventilation compared to the small stimulation of minute ventilation seen during aerial hypercapnia. Some animals responded to aquatic hypercapnia with a series of three or four closely spaced breaths separated by long nonventilatory periods. Overall, T. natans showed little capacity to modulate expiratory or inspiratory tidal volumes and depended heavily on changing air-breathing frequency to meet hypoxic and hypercapnic challenges. These responses are different from those of anurans or urodeles studied to date, which modulate both the number of ventilatory oscillations in lung-inflation cycles and the degree of lung inflation when challenged with peripheral or central chemoreceptor stimulation. PMID- 10685904 TI - Maximum daily energy intake: it takes time to lift the metabolic ceiling. AB - Conventionally, maximum capacities for energy assimilation are presented as daily averages. However, maximum daily energy intake is determined by the maximum metabolizable energy intake rate and the time available for assimilation of food energy. Thrush nightingales (Luscinia luscinia) in migratory disposition were given limited food rations for 3 d to reduce their energy stores. Subsequently, groups of birds were fed ad lib. during fixed time periods varying between 7 and 23 h per day. Metabolizable energy intake rate, averaged over the available feeding time, was 1.9 W and showed no difference between groups on the first day of refueling. Total daily metabolizable energy intake increased linearly with available feeding time, and for the 23-h group, it was well above suggested maximum levels for animals. We conclude that both intake rate and available feeding time must be taken into account when interpreting potential constraints acting on animals' energy budgets. In the 7-h group, energy intake rates increased from 1.9 W on the first day to 3.1 W on the seventh day. This supports the idea that small birds can adaptively increase their energy intake rates on a short timescale. PMID- 10685905 TI - Seasonal thermogenic acclimation of diurnally and nocturnally active desert spiny mice. AB - Diurnally active golden spiny mice (Acomys russatus) and nocturnal common spiny mice (Acomys cahirinus) coexist in hot rocky deserts of Israel. Diurnal and nocturnal activities expose these species to different climatic conditions. Nonshivering thermogenesis (NST) capacity of individuals of both species immediately upon removal from the field exhibited seasonal changes, with no significant interspecific difference. Colony-reared mice of either species transferred in the laboratory from long to short photoperiod increased NST capacity, though to a lesser extent than observed in the seasonal acclimatization. The underlying biochemical mechanisms of short photoperiod acclimation differed between the species. In both Cytochrome-c oxidase (Cox) activity was higher in short as compared to long photoperiod. In short photoperiod-acclimated A. cahirinus uncoupling protein (UCP) content in brown adipose tissue (BAT) was significantly higher than in long photoperiod, while in A. russatus there was no significant change. In A. russatus there was a significant increase in lipoprotein lipase (LPL) activity in BAT in short photoperiod-acclimated individuals, while in A. cahirinus LPL activity was high under both acclimations. The low LPL activity in brown adipose tissue of desert adapted A. russatus may facilitate lipid uptake in white adipose tissue, an advantage in desert conditions where food is scarce and irregularly distributed in space and time. PMID- 10685906 TI - Effect of a low-Fat diet on body composition and blubber fatty acids of captive juvenile harp seals (Phoca groenlandica). AB - We investigated the effects of a change from a high-fat diet to a low-fat diet of differing fatty acid (FA) composition on the body composition and blubber FA of five captive juvenile harp seals. Seals that had been maintained for 1 yr on a diet of Atlantic herring (>/=9% fat) were switched to a diet of Atlantic pollock (1. 7% fat) for 30 d. On days 0, 14, and 30, mass and body composition (using isotope dilution) were measured, and blubber biopsies (5 cmx6 mm) were taken for FA analysis. Fat accounted for 38%-49% of body mass at the start of the experiment. When switched to the pollock diet, and despite food intakes averaging 6.5 kg/d (32.3 MJ/d), body fat declined by an average of 6.4 kg or by 32% over the 30-d experiment. In contrast, body protein increased in direct relation to protein intake (r2=0.836, P=0.030). Despite substantial loss of body fat, blubber FA signature changed significantly to reflect the changes in dietary intake of FA, and the deposition of FA was quantifiably predictable. Our results suggest that young growing phocids are unable to maintain body fat stores on low-fat diets even when protein intakes are high. This may have significant implications for juvenile pinniped survival in the wild. In addition, turnover and deposition of dietary FA in blubber takes place in nonfattening seals. PMID- 10685907 TI - Effects of electromagnetic fields on the reproductive success of American kestrels. AB - Reduced reproductive success of birds nesting near power lines has been documented but never directly attributed to electromagnetic fields (EMFs). Laboratory studies have identified EMF effects on embryonic development, but reproductive success of wild birds is dependent on additional factors, including fertility, egg size, hatching, and fledging success. We tested whether EMFs affect reproductive success of birds. Captive American kestrels (Falco sparverius) were bred for one season per year for 2 yr under either controlled or EMF conditions. EMF exposure was equivalent to that experienced by wild reproducing kestrels and was weakly associated with reduced egg laying in 1 yr only. In both years fertility was higher, but hatching success was lower in EMF pairs than control pairs. Fledging success was higher in EMF pairs than control pairs in 1995 only. Egg composition and embryonic development were examined in 1 yr only, but hatchlings were measured in both years. EMF eggs were larger, with more yolk, albumen, and water, but had thinner egg shells than control eggs. Late term EMF embryos were larger and longer than control embryos, although hatchlings were similar in body mass and size. EMF exposure affected reproductive success of kestrels, increasing fertility, egg size, embryonic development, and fledging success but reducing hatching success. PMID- 10685908 TI - Effects of diet on titratable acid-base excretion in grasshoppers. AB - Despite the potential for diet to affect organismal acid-base status, especially in herbivores, little is known about the effects of diet on acid-base loading and excretion. We tested the effects of diet on acid-base loading and excretion in grasshoppers by (a) comparing the fecal acid-base content of 15 grasshopper species collected from the field and (b) comparing fecal acid-base excretion rates of Schistocerca americana grasshoppers fed vegetable diets that differed in their ashed and raw acid-base contents. The field experiments indicated that grass-feeding species excrete fairly neutral fecal pellets, while forb/mixed feeding species vary widely in their fecal acid-base contents. In the laboratory experiment, acid-base excretion rates were positively correlated with dietary ashed base intake rates but were not correlated with the acid-base content of raw, unashed diet or feeding rate. These experiments suggest that some diets could strongly challenge the acid-base homeostasis of herbivores; in some grasshoppers, dietary acid-base loads could produce certainly lethal 1-unit changes in average body pH within 6 h if they were not excreted. PMID- 10685909 TI - Respiratory adaptations to running-water microhabitats in mayfly larvae Epeorus sylvicola and Ecdyonurus torrentis, Ephemeroptera. AB - The mayfly larvae Epeorus sylvicola and Ecdyonurus torrentis inhabit either fast flowing or, for the latter species, calm zones of running water. We studied (1) mechanisms and limitations of oxygen transport in single individuals (oxygen consumption rate, occurrence and rate of gill movements, and heartbeats) in running water of different oxygen concentrations and (2) capacities for anaerobiosis (L-lactate production). Our aim was to look for specific adaptations in the two species to slightly different microhabitats. Epeorus sylvicola, whose immovable gills are not able to generate ventilatory convection, proved to be an oxyconformer at both test temperatures (11 degrees and 15 degrees C). Ecdyonurus torrentis showed a progressively stronger oxyregulatory behavior at higher temperatures. In this species an onset of gill beating was found at moderate hypoxia (below 16 kPa). Ventilating individuals reached maximum rates (300 min-1) of 5-14 kPa. In the case of a further reduction of oxygen partial pressure, the ventilatory rate started to decrease. Ventilatory activity, however, was maintained down to very low oxygen concentrations. Neither in E. sylvicola nor in E. torrentis was experimental evidence found to confirm the hypothesis of a respiratory function of hindgut movements. During hypoxia, the heart rate was constant in both species (E. sylvicola: 80 min-1; E. torrentis: 60 min-1): bradycardia occurred either below 1.5 kPa or below 4 kPa. Anaerobiosis, that is, lactate production, was not detected in either species. PMID- 10685910 TI - Evolutionary history and adaptive significance of respiratory structures on the legs of intertidal porcelain crabs, genus Petrolisthes. AB - Semiterrestrial and terrestrial crabs have evolved multiple strategies for aerial respiration. An uncommon strategy for aerial respiration is seen in porcelain crabs, genus Petrolisthes, where decalcified areas on the meral segments of the walking legs are used as respiratory structures. Here, the evolutionary history and adaptive significance of these structures in porcelain crabs is examined. Interspecific variation in leg membrane size is from 0% to 60% of the surface area of the meral segment. Leg membrane relative size is positively correlated with body size across species but not within one species, Petrolisthes cinctipes. Phylogenetic analyses suggest that leg membranes are ancestral to one of two eastern Pacific Petrolisthes clades. Comparative analyses using phylogenetic independent contrasts indicate a relationship between leg membrane relative size and body size that is phylogenetically independent. In large-bodied intertidal species, whole-animal lactate accumulation during aerial incubation is 200%-300% higher when the leg membranes are obscured, indicating that the leg membranes are functional respiratory structures in these species. Thus, it is possible that leg membranes have facilitated the evolution of larger body sizes by providing additional respiratory surfaces to accommodate the associated higher metabolic demands. PMID- 10685911 TI - Diet explains interpopulation variation of plasma carotenoids and skin pigmentation in nestling white storks. AB - Carotenoids have a dietary origin in birds, but mechanisms by which they are absorbed in the gut, transported in the blood, metabolized at various sites, and deposited in the integument remain poorly understood. Variation in both plasma carotenoid levels and external color may reflect different access to dietary carotenoids or individual physiological differences in the uptake and deposition of carotenoids. We compared total plasma carotenoid concentration in nestling white storks (Ciconia ciconia) from 11 Spanish colonies in two consecutive years. The main food item in one of the colonies was the red swamp crayfish (Procambarus clarkii), a recently introduced species. Storks in the remaining colonies ate a variety of foods but no crayfish. Total plasma carotenoid levels in the colony where crayfish were consumed were about five times higher than in any other colony. These differences were maintained after controlling for the significant interyear variability, as well as for sex, age, and body mass of birds. Skin pigmentation also differed, being intensely orange in storks that consumed crayfish but white (unpigmented) in the remaining individuals. With thin-layer chromatography (TLC) and electronic absorption spectroscopy, astaxanthin was confirmed as the major carotenoid in crayfish as well as in the plasma, skin, and body fat of crayfish-eating storks, whereas lutein was the main carotenoid in plasma samples from the other colonies. These results indicate that a newly available carotenoid in the environment, astaxanthin, can be absorbed in large quantities from the gut and be transported in the blood before deposition in different tissues. PMID- 10685912 TI - The energy metabolism of common carp (Cyprinus carpio) when exposed to salt stress: an increase in energy expenditure or effects of starvation? AB - Stenohaline common carp (Cyprinus carpio) were chronically exposed to the two main osmoregulatory ions, Na+ and Cl-, at levels close to their isoosmotic value for 28 d (171 mM NaCl; 324 mosm kg-1; 10 per thousand). The aim of this study was to assess whether or not the disturbed ion and osmoregulation affected the energy demand and the energy stores of the exposed fish. Salt exposure reduced food intake by 70% and had adverse effects on growth and survival. Although food consumption decreased and growth was seriously affected, routine oxygen consumption of the exposed fish did not drop, indicating a reallocation of energy expenditure from growth toward other processes. A stress-induced increase in plasma glucose was observed. As a result of low food intake, lower levels of protein were used for fuel. Protein use itself was probably replaced by the use of carbohydrates. These effects were confirmed by the depletion of both muscle and liver glycogen stores during the experimental period. We conclude that, besides the effects of reduced feeding, stress induced extra energy requirements leading to the depletion of energy stores. PMID- 10685913 TI - Effects of body mass and reproduction on the basal metabolic rate of brown long eared bats (Plecotus auritus). AB - We measured basal metabolic rate (BMR) of nonreproductive and of breeding (pregnant and lactating) female brown long-eared bats (Plecotus auritus) to investigate the effects of intra- and interindividual variation in body mass and of reproduction on metabolism. The BMR of six nonreproductive females was measured between five and seven times at approximately 2-wk intervals over a period of 2.5 mo. There was a highly significant effect (P<0.001) of body mass on BMR of these nonreproductive females. The pooled within-individual scaling exponent (1.88) significantly exceeded the established mammalian interspecific exponent (0.75). In addition, we made single observations on 14 nonreproductive females to establish the effects of differences in mass between individuals. The mean BMR across all 14 individuals was 82 mW (+/-24 SD). There was a significant positive relationship between BMR and body mass across these individuals (r2=0.39), with a between-individual scaling exponent of 0.75. Inter- and intraindividual effects of mass on BMR were combined in a regression analysis that included mean body mass and deviation from mean mass on any given day as predictors. This regression model explained 55% of the variation in BMR. We made longitudinal measurements of BMR throughout reproduction and compared these with the predicted BMR of nonreproductive bats of the same body mass. Reproductive females exhibited temporal flexibility in BMR. BMR during pregnancy increased on a whole-animal basis but was significantly lower (by, on average, 15%) than BMR predicted for nonreproductive females of the same mass. Over a period of 1-75 d following birth, whole-animal BMR was greater than that during pregnancy, even though body mass declined after parturition. Hence, postbirth BMR was greater than the level predicted for nonreproductive females of the same mass. This study indicates that the scaling of BMR with body mass differs significantly within and between individuals and that there is a reduction of BMR in pregnancy and an elevation of BMR during lactation. PMID- 10685914 TI - How to choose delivery devices for asthma. PMID- 10685915 TI - Pectus excavatum: studiously ignored in the United Kingdom? PMID- 10685916 TI - Public health: Establishing an interagency equipment fund for children with disabilities. PMID- 10685917 TI - Stamps in pediatrics: Thalassaemia. PMID- 10685918 TI - Controversy: Is the Children Act failing severely abused and neglected children? PMID- 10685919 TI - Is routine growth monitoring effective? A systematic review of trials. AB - BACKGROUND: Growth monitoring consists of routine measurements to detect abnormal growth, combined with some action when this is detected. It aims to improve nutrition, reduce the risk of death or inadequate nutrition, help educate carers, and lead to early referral for conditions manifest by growth disorders. As primary care workers world wide invest time in this activity, evidence for its benefits and harms was sort. INCLUSION CRITERIA: STUDIES: randomised or quasi randomised controlled trials of growth monitoring. INTERVENTIONS: regular growth monitoring, combined with some intervention targeted at abnormal growth, compared with controls. OUTCOMES: anthropometric measures; referrals to primary and specialist care, or community services; maternal knowledge, anxiety, and satisfaction; child morbidity and mortality. COMPARISONS: Routine growth monitoring compared with no routine growth monitoring; routine growth monitoring by plotting onto a standard chart compared with monitoring with no chart. SEARCH STRATEGY: Cochrane controlled trials register; World Health Organisation and World Bank publications; contact with specialist community paediatricians working in the field. RESULTS: Two trials met the inclusion criteria. One compared growth monitoring with no growth monitoring, in a cluster randomised trial nested in a nutritional intervention programme, and detected no difference in nutritional outcomes between the two groups. Another trial compared growth monitoring with and without a standard chart, measuring maternal knowledge of women about nutrition. It showed small numerical differences in test scores. DISCUSSION AND IMPLICATIONS: Current policies appear to be based on the opinion that investment in the activity has worthwhile health benefits, and does no harm. No reliable evidence was found to support or refute this. PMID- 10685920 TI - Parental recall of birth weight: how accurate is it? AB - OBJECTIVE: To assess the accuracy of parental recall of birth weight in a British population and to investigate whether social class and age of the child significantly influence the accuracy of recalled birth weight. METHODS: A questionnaire was given to parents whose children were participating in a blood pressure study and the hospital records were retrieved to check the birth weight data. RESULTS: At the time of the study, the children (n = 649) ranged in age from 6 to 15 years. Seventy five per cent of the recalled birth weights were within 50 g of that recorded in the hospital records. No significant associations were found between the difference in birth weights (recalled birth weight minus hospital record) and social class of the parents or age of the child at time of data collection. CONCLUSION: This large study shows that parental recall of birth weight is good across the social classes and up to 16 years after delivery. There was no evidence of systematic bias, which would prejudice results of studies on the relation of birth weight to adult hypertension. PMID- 10685921 TI - Height and weight reference curves for homozygous sickle cell disease. AB - OBJECTIVE: To derive height and weight growth reference curves for children with homozygous sickle cell disease. STUDY DESIGN: Subjects (n = 315) were participants in a population based, longitudinal cohort study of sickle cell disease in Kingston, Jamaica. Regular measurements of height and weight were made from enrollment into the study at birth up to 22 years of age. RESULTS: Sex specific growth reference curves for height for age and weight for age covering the age range 0-18 years are presented. CONCLUSION: These growth reference curves are suitable for identifying coincidental growth problems in children with homozygous sickle cell disease. PMID- 10685922 TI - Wechsler subscale IQ and subtest profile in early treated phenylketonuria. AB - AIM: Mildly depressed IQ is common in treated phenylketonuria. This study explored whether a particular intellectual ability profile typifies early and continuously treated phenylketonuria and whether component skills comprising the IQ relate to socioeconomic and treatment factors. METHODS: IQ scores were collected retrospectively from variants of the "Wechsler intelligence scale for children" performed at age 8 on 57 children with early treated, classic phenylketonuria. The mental ability pattern underlying IQ was investigated by analysing subscale and subtest scores and dietary factors, such as historical phenylalanine blood concentrations. RESULTS: The children's mean full scale IQ of 91.11 was significantly below the healthy population norm. There was a significant discrepancy between their mean verbal IQ (94.65) and mean performance IQ (89.42), suggestive of a spatial deficit, but the data did not support a biochemical or sociological explanation. Individual Wechsler subtests had no distinctive pattern. Phenylalanine control at age 2 was predictive of overall IQ. At this age, children with annual median phenylalanine < 360 micromol/litre (recommended UK upper limit) had a mean IQ 10 points higher than those above. CONCLUSIONS: Early and continuous treatment of phenylketonuria does not necessarily lead to normalisation of overall IQ. Verbal intelligence in the primary school years appears to normalise if blood phenylalanine is maintained below 360 micromol/litre in infancy, but spatial intelligence may remain poor. However, the discrepancy in skill development is not the result of social status or treatment variables. Perhaps weak spatial intelligence is an ancillary effect of a protective rearing style occasioned by the dietary treatment regimen. PMID- 10685923 TI - The limping child. PMID- 10685924 TI - Effect of genotype on changes in intelligence quotient after dietary relaxation in phenylketonuria and hyperphenylalaninaemia. AB - BACKGROUND: Associations between genotype and intellectual outcome in patients with phenylketonuria are complicated because intelligence is influenced by many variables, including environmental factors and other genetic determinants. Intellectual changes with age, both on and after relaxation of diet, vary within the patient population. This study aims to determine whether a significant association exists between genotype and change in intelligence after relaxation of diet. METHODS: 125 patients with hyperphenylalaninaemia and phenylketonuria whose diet was relaxed after 8 years of age. Verbal, performance, and full scale intelligence quotients at 8, 14, and 18 years were expressed as standard deviation scores (IQ-SDS), and genotype as predicted residual enzyme activity (PRA) of phenylalanine hydroxylase. RESULTS: IQ-SDS at 8, 14, and 18 years were significantly below normal; no association was found between PRA and IQ-SDS. Significant reductions in verbal and full scale IQ-SDS occurred between 8 and 14 years and 8 and 18 years. There was a significant association between PRA and the reduction in verbal, performance, and full scale IQ between these years. Multiple regression analysis of 18 year results, using 8 year results as covariates, supported the association between PRA and IQ-SDS; after adjustment for phenylalanine control, both up to and after the age of 8 years, the full scale IQ SDS at 14 and 18 years was 0.15 higher for each 10% increase in PRA. CONCLUSIONS: Genotype might be useful in predicting the likelihood of intellectual change in patients with hyperphenylalaninaemia and phenylketonuria whose diet is relaxed after the age of 8 years. PMID- 10685925 TI - Behavioural phenotypes: what do they teach us? PMID- 10685926 TI - Diagnosing idiopathic/cryptogenic epilepsy syndromes in infancy. AB - PURPOSE: To determine the characteristics that permit diagnosis of the type of epilepsy beginning in the 1st year of life, and to determine from what age such characteristics are recognisable. PATIENTS: From 430 non-selected patients who began having seizures in the 1st year of life and were referred to the neuropaediatric department of Saint Vincent de Paul Hospital, those with epileptic spasms as the first seizure type, those with recognisable aetiology, and those for whom early history was not reliable were excluded. METHODS: For the remaining 140 patients, the age at which clinical and electroencephalogram (EEG) characteristics met those of recognisable epilepsy syndromes according to the ILAE classification was studied. RESULTS: In most epilepsy syndromes, the diagnosis could be made within three months of onset of the disorder. The most difficult was to distinguish cryptogenic localisation related epilepsy from severe myoclonic epilepsy in infancy. Repeat focal seizures and persisting spike focus were the earliest and most reliable signs of localisation related epilepsy, whereas alternating focal seizures, generalised myoclonus, and/or spike waves appeared before the end of the 1st year in most infants with severe myoclonic epilepsy. However, for 39 patients it was not possible to reach the diagnosis of a precise syndrome. CONCLUSION: For over three quarters of infants with cryptogenic/idiopathic epilepsy, it is possible to reach a syndromic diagnosis within the first months of the disease, based on clinical and EEG characteristics. However, for one quarter, no diagnosis is possible based on the currently available classification. PMID- 10685927 TI - Morbidity in reflex sympathetic dystrophy. AB - Reflex sympathetic dystrophy (RSD), an unusual diagnosis in general paediatrics, is well recognised by paediatric rheumatologists. This study reports the presentation and the clinical course of 46 patients (35 female, age range 8-15.2) with RSD. The patients saw professionals from an average of 2.3 specialties (range 1-5). Twenty five (54%) had a history of trauma. Median time to diagnosis was 12 weeks (range 1-130). Many children had multiple investigations and treatments. Once diagnosis was made, treatment followed with physiotherapy and analgesics. Median time to recovery was seven weeks (range 1-140), with 27.5% relapsing. Nine children required assessment by the child and adolescent psychiatry team. This disease, though rare, has significant morbidity and it is therefore important to raise clinicians' awareness of RSD in childhood. Children with the condition may then be recognised and referred for appropriate management earlier, and spared unnecessary investigations and treatments which may exacerbate the condition. PMID- 10685928 TI - Proteus syndrome and immunodeficiency. AB - A 10 year old boy with Proteus syndrome presented with a pericardial effusion of unknown aetiology. Immunological investigation revealed low serum IgG and IgA, accompanied by low levels of specific antibodies to pneumococcal and haemophilus type B polysaccharides. Circulating lymphocyte surface marker profile revealed T and B cell lymphopenia. This is the first report of hypogammaglobulinaemia occurring in the Proteus syndrome. PMID- 10685929 TI - Fits, pyridoxine, and hyperprolinaemia type II. AB - The rare inherited disorder hyperprolinaemia type II presents with fits in childhood, usually precipitated by infection. A diagnosis of hyperprolinaemia type II and vitamin B(6) deficiency was made in a well nourished child with fits. It is thought that pyridoxine deficiency was implicated in her fits and was the result of inactivation of the vitamin by the proline metabolite, pyrroline-5 carboxylate. PMID- 10685930 TI - Physical treatment of fever. AB - Fever is a common symptom of childhood illness, and much time and effort is spent in the pursuit of reducing high temperature. Although antipyretic drugs are the main form of treatment, this report considers the part that physical treatments might play in reducing the temperature of febrile children. Such treatments include tepid sponging, removing clothing, and cooling the environment. Of these treatments, tepid sponging has been studied most extensively, as an addition to paracetamol, but seems to offer little advantage over paracetamol alone. It is likely that other methods might be equally ineffective because they all rely on similar methods of heat loss. PMID- 10685931 TI - Laparoscopic surgery in children. PMID- 10685932 TI - Concentrations of antimony in infants dying from SIDS and infants dying from other causes. AB - OBJECTIVES: Raised concentrations of antimony have been found in infants dying of sudden infant death syndrome (SIDS). The presumed source of this antimony is toxic gases generated from fire retardants that are present in cot mattresses. The aim of this study was to determine the role of antimony in SIDS. DESIGN: Samples of liver, brain, serum, and urine were collected from all patients dying from SIDS and a group of aged matched control infants who had died of other causes. SETTING: Nationwide study in Ireland. SUBJECTS: 52 infants dying from SIDS and 19 control infants aged > 7 days and < 1 year. RESULTS: The median concentration of antimony in the liver and brain of infants dying of SIDS was < 1 ng/g, with no difference detected between the infants dying from SIDS and the control infants. The range of antimony in the serum of infants dying of SIDS was 0.09-0.71 microg/litre (median, 0.26). Although no difference was found between infants dying from SIDS and control infants, SIDS infants were found to have higher concentrations when compared with healthy infants in the 1st year of life, probably as a result of release of antimony into serum after death. Urine antimony concentrations in infants dying from SIDS were < 3.91 ng/mg (corrected for creatinine) and similar to values found both in control infants and healthy infants. CONCLUSION: There is no evidence to support a causal role for antimony in SIDS. PMID- 10685933 TI - Infant feeding and adult glucose tolerance, lipid profile, blood pressure, and obesity. AB - BACKGROUND: It is generally accepted that breast feeding has a beneficial effect on the health of infants and young children. Recently, a few studies have shown that the method of infant feeding is also associated with cardiovascular disease and its risk factors in adult life. AIMS: To examine the association between the method of infant feeding in the first weeks after birth and glucose tolerance, plasma lipid profile, blood pressure, and body mass in adults aged 48-53 years. METHODS: Subjects born at term between 1 November 1943 and 28 February 1947 in the Wilhelmina Gasthuis in Amsterdam around the time of a severe period of famine (late November 1944 to early May 1945). For 625 subjects, information was available about infant feeding at the time of discharge from hospital (on average 10.4 days after birth), and at least one blood sample after an overnight fast. RESULTS: Subjects who were bottle fed had a higher mean 120 minute plasma glucose concentration after a standard oral glucose tolerance test than those who were exclusively breast fed. They also had a higher plasma low density lipoprotein (LDL) cholesterol concentration, a lower high density lipoprotein (HDL) cholesterol concentration, and a higher LDL/HDL ratio. Systolic blood pressure and body mass index were not affected by the method of infant feeding. CONCLUSIONS: Exclusive breast feeding seems to have a protective effect against some risk factors for cardiovascular disease in later life. PMID- 10685934 TI - Effect of prone sleeping on circulatory control in infants. AB - BACKGROUND: The mechanism of death in sudden infant death syndrome (SIDS) remains unclear. Progressive bradycardia is the pre-eminent terminal event, suggesting that circulatory failure might be a crucial factor. Vasomotor tone regulates the circulatory system by controlling blood volume distribution while maintaining venous return and blood pressure. AIM: To examine whether prone sleeping, the most consistently identified risk factor for SIDS, has a measurable influence on vasomotor/circulatory control. METHODS: 44 full term infants (mean age, 7.9 weeks) were studied during an overnight sleep. Recordings were made while the infants were horizontal and asleep in the supine and prone positions, and repeated after a head up tilt to 60 degrees, maintained for 30 minutes, while in both sleep positions. Blood pressure, heart rate, anterior shin, and anterior abdominal wall skin temperatures were measured. RESULTS: Systolic blood pressure was lower, but peripheral skin temperature and heart rate were higher during sleep, while horizontal, in the prone rather than the supine position. After tilting, there was a greater reduction in blood pressure and a greater increase in peripheral skin temperature and heart rate when in the prone position. Anterior abdominal wall skin temperature did not vary in either sleeping positions while horizontal or tilted. CONCLUSION: Prone sleeping has a measurable effect on circulatory control, with a reduction in vasomotor tone resulting in peripheral vasodilatation, a higher peripheral skin temperature, a lower blood pressure, and a higher resting heart rate. Because vasomotor tone is crucially important in circulatory control this could be a factor in increasing the risk of SIDS. PMID- 10685935 TI - Temporal bone computed tomography findings in bilateral sensorineural hearing loss. AB - AIM: To examine the yield of computed tomography (CT) of the temporal bones when investigating sensorineural hearing loss (SNHL) and to identify factors associated with CT findings. METHODS: Retrospective analysis of 116 consecutively investigated children with bilateral SNHL at the audiology department of Great Ormond Street Hospital, London. Main outcome measures were CT results, hearing loss parameters, history, and clinical examination. RESULTS: A total of 33 (28.4%) CT scans were identified as abnormal. Children with profound and/or progressive hearing loss and/or craniofacial abnormalities were more likely to have an abnormal CT scan and together accounted for 25 abnormal CT scans. Sex, consanguineous parents, or family history of SNHL were not associated with CT findings. Dilated vestibular aqueduct was significantly correlated with the presence of progressive SNHL. CONCLUSIONS: All children with SNHL should undergo radiological investigation of the petrous bones/inner ear; abnormalities are more likely to be found in cases with craniofacial abnormalities, or profound or progressive hearing loss. The decision whether to perform a CT or magnetic resonance imaging will depend on scanner availability, expertise, and management considerations, but cochlear implant candidates will require both. PMID- 10685936 TI - Measurement and interpretation of blood pressure. PMID- 10685937 TI - FETAL AND NEONATAL EDITION march 2000 issue PMID- 10685979 TI - Genetics of neonatal hyperinsulinism. AB - Congenital hyperinsulinism (HI) is a clinically and genetically heterogeneous entity. The clinical heterogeneity is manifested by severity ranging from extremely severe, life threatening disease to very mild clinical symptoms, which may even be difficult to identify. Furthermore, clinical responsiveness to medical and surgical management is extremely variable. Recent discoveries have begun to clarify the molecular aetiology of this disease and thus the mechanisms responsible for this clinical heterogeneity are becoming more clear. Mutations in 4 different genes have been identified in patients with this clinical syndrome. Most cases are caused by mutations in either of the 2 subunits of the beta cell ATP sensitive K(+) channel (K(ATP)), whereas others are caused by mutations in the beta cell enzymes glucokinase and glutamate dehydrogenase. However, for as many as 50% of the cases, no genetic aetiology has yet been determined. The study of the genetics of this disease has provided important new information about beta cell physiology. Although the clinical ramifications of these findings are still limited, in some situations genetic studies might greatly aid in patient management. PMID- 10685980 TI - Hyperinsulinism of infancy: towards an understanding of unregulated insulin release. European Network for Research into Hyperinsulinism in Infancy. AB - Insulin is synthesised, stored, and secreted from pancreatic beta cells. These are located within the islets of Langerhans, which are distributed throughout the pancreas. Less than 2% of the total pancreas is devoted to an endocrine function. When the mechanisms that control insulin release are compromised, potentially lethal diseases such as diabetes and neonatal hypoglycaemia are manifest. This article reviews the physiology of insulin release and illustrates how defects in these processes will result in the pathophysiology of hyperinsulinism of infancy. PMID- 10685981 TI - Practical management of hyperinsulinism in infancy. AB - Hyperinsulinism in infancy is one of the most difficult problems to manage in contemporary paediatric endocrinology. Although the diagnosis can usually be achieved without difficulty, it presents the paediatrician with formidable day to day management problems. Despite recent advances in understanding the pathophysiology of hyperinsulinism, the neurological outcome remains poor, and there is often a choice of unsatisfactory treatments, with life long sequelae for the child and his or her family. This paper presents a state of the art overview on management derived from a consensus workshop held by the European network for research into hyperinsulinism (ENRHI). The consensus is presented as an educational aid for paediatricians and children's nurses. It offers a practical guide to management based on the most up to date knowledge. It presents a proposed management cascade and focuses on the clinical recognition of the disease, the immediate steps that should be taken to stabilise the infant during diagnostic investigations, and the principles of definitive treatment. PMID- 10685982 TI - Persistent hyperinsulinaemic hypoglycaemia of infancy: a heterogeneous syndrome unrelated to nesidioblastosis. PMID- 10685983 TI - Maternal insulin-like growth factor binding protein-1, body mass index, and fetal growth. AB - AIM: To examine the hypothesis that the maternal insulin-like growth factor system may constrain fetal growth. METHODS: A prospective observational study of maternal serum insulin-like growth factor binding protein-1 (IGFBP-1) and fetal growth was undertaken in neonates with birthweights below the 5th centile. They had been classified either as having fetal growth restriction (FGR) due to placental dysfunction (increased umbilical artery Doppler pulsatility index (PI); n = 25) or as being small for gestational age (SGA; normal umbilical artery PI, growth velocity and amniotic fluid; n = 27). Eighty nine controls had normal birthweights (5th-95th centile), umbilical artery PI, growth velocity, and amniotic fluid. IGFBP-1 was measured by radioimmunoassay. RESULTS: Among the controls, there was no significant correlation between IGFBP-1 and birthweight after allowing for body mass index (BMI). Maternal BMI was high in FGR and after adjusting for this, IGFBP-1 was increased (109 ng/ml) compared with SGA babies (69 ng/ml) and controls (57 ng/ml) and correlated with the umbilical artery PI. CONCLUSIONS: Maternal IGFBP-1 is probably not part of normal placental function. Its increase in FGR could be the cause or consequence of impaired placental perfusion, but high IGFBP-1 concentrations might further reduce the availability of maternal IGF-I to the placenta. This could worsen placental function and so adversely affect fetal growth. PMID- 10685984 TI - Risk adjusted and population based studies of the outcome for high risk infants in Scotland and Australia. International Neonatal Network, Scottish Neonatal Consultants, Nurses Collaborative Study Group. AB - OBJECTIVES: To compare outcomes of care in selected neonatal intensive care units (NICUs) for very low birthweight (VLBW) or preterm infants in Scotland and Australia (study 1) and perinatal care for all VLBW infants in both countries (study 2). DESIGN: Study 1: risk adjusted cohort study; study 2: population based cohort study. SUBJECTS: Study 1: all 2621 infants of < 1500 g birth weight or < 31 weeks' gestation admitted to a volunteer sample of hospitals comprising eight of all 17 Scottish NICUs and six of all 12 tertiary NICUs in New South Wales and Queensland in 1993-1994; study 2: all 5986 infants of 500-1499 g birth weight registered as live born in Scotland and Australia in 1993-1994. MAIN OUTCOMES: Study 1: (a) hospital death; (b) death or cerebral damage, each adjusted for gestation and CRIB (clinical risk index for babies); study 2: neonatal (28 day) mortality. RESULTS: Study 1. Data were obtained for 1628 admissions in six Australian NICUs, 775 in five Scottish tertiary NICUs, and 148 in three Scottish non-tertiary NICUs. Crude hospital death rates were 13%, 22%, and 22% respectively. Risk adjusted hospital mortality was about 50% higher in Scottish than in Australian NICUs (adjusted mortality ratio 1.46, 95% confidence interval (CI) 1.29 to 1.63, p < 0.001). There was no difference in risk adjusted outcomes between Scottish tertiary and non-tertiary NICUs. After risk adjustment, death or cerebral damage was more common in Scottish than Australian NICUs (odds ratio 1.9, 95% CI 1.5 to 2.5). Both these risk adjusted adverse outcomes remained more common in Scottish than Australian NICUs after excluding all infants < 28 weeks' gestation from the comparison. Study 2. Population based neonatal mortality in infants of 500-1499 g was higher in Scotland (20.3%) than Australia (16.6%) (relative risk 1.22, 95% CI 1.08 to 1. 39, p = 0.002). In a post hoc analysis, neonatal mortality was also higher in England and Wales than in Australia. CONCLUSIONS: Study 1: outcome was better in the Australian NICUs. Study 2: perinatal outcome was better in Australia. Both results may be consistent, at least in part, with differences in the organisation and implementation of neonatal care. PMID- 10685985 TI - Reproductive decisions after neonatal screening identifies cystic fibrosis. AB - AIMS: To document the reproductive choices made by women in New South Wales, Australia, after neonatal screening has identified cystic fibrosis (CF). METHODS: A sample of women attending cystic fibrosis clinics in New South Wales who had a child (or children) diagnosed by neonatal screening between 1981 and 1996 were interviewed. RESULTS: Two thirds of the women chose to avoid having another child with CF. The uptake of prenatal diagnosis was 66% in women who had a subsequent pregnancy; of these 69% terminated or would have terminated an affected fetus. Fifty nine per cent of the women who decided against a further pregnancy made this decision in order to avoid having another child with CF. CONCLUSIONS: These data show that having a child with CF influenced subsequent reproductive choices. In addition to the medical advantages of an early diagnosis offered by neonatal screening, this also allows informed future reproductive decisions. PMID- 10685986 TI - Cranial ultrasound abnormalities in full term infants in a postnatal ward: outcome at 12 and 18 months. AB - OBJECTIVE: To investigate whether cranial ultrasound abnormalities found in low risk full term infants had any influence on neurodevelopmental outcome. METHODS: For 103 infants who had a neurological assessment, a cranial ultrasound examination, and for whom antenatal and perinatal data were collected within 48 hours of delivery, neurodevelopmental status was evaluated at 12 and 18 months. The results of a scored neurological examination and the Griffiths mental developmental scale were correlated with the presence and type of ultrasound abnormality found in the neonatal period. RESULTS: None of the infants with ultrasound abnormalities showed any signs of cerebral palsy or severe developmental delay. There was also no significant difference between the overall neurological and neurodevelopmental scores of the infants with normal and abnormal ultrasound findings. However, when the individual subscales of the Griffiths test were analysed, all infants with bulky choroid or intraventricular haemorrhage had normal scores in all subscales, four of eight with periventricular white matter lesions had low scores on the locomotor subscale, and three of five with asymmetrical ventricles had low scores on the performance subscale. The presence of adverse antenatal and perinatal factors did not affect the outcome in this group. CONCLUSION: Incidental ultrasound abnormality in full term neonates, in particular intraventricular haemorrhage, although common, appear to have a good prognosis. Longer follow up studies are needed to see whether some of these infants, in particular those with white matter lesions, develop dyspraxia or other minor neurological impairments at school age. PMID- 10685987 TI - Correlation between visual function, neurodevelopmental outcome, and magnetic resonance imaging findings in infants with periventricular leucomalacia. AB - AIM: To evaluate the correlation between visual function and neurodevelopmental outcome in children with periventricular leucomalacia at 1 and 3 years. METHOD: Visual acuity, visual field, ocular motility, and optokinetic nystagmus were tested in 29 infants with periventricular leucomalacia by brain magnetic resonance imaging. All infants also had a structured neurological examination and a Griffiths developmental assessment. RESULTS: 21 of the infants showed at least one abnormality of visual function. The degree of visual impairment-that is, the number of visual tests showing abnormal results-correlated well with the results on developmental assessment at both ages. CONCLUSION: Multivariate analysis showed that visual impairment was the most important variable in determining the neurodevelopmental scores of these infants, more than their motor disability and the extent of their lesions on magnetic resonance imaging. PMID- 10685988 TI - Sensorineural hearing loss and prematurity. AB - OBJECTIVE: To elucidate clinical antecedents of sensorineural hearing loss (SNHL) in very preterm infants. DESIGN: Case-control study. SUBJECTS: Fifteen children < 33 weeks' gestation with significant SNHL born between 1 January 1990 and 31 December 1994, detected within 9 months of birth, and 30 matched control children. METHODOLOGY: Perinatal variables in the two groups were compared using non-parametric tests and conditional logistic regression (EGRET). RESULTS: Median birth weight for the index group was 960 g (range 600-2914 g) compared with 1026 g (range 410-2814 g) for controls. Children with SNHL had longer periods of intubation, ventilation, oxygen treatment, and acidosis, and more frequent treatment with dopamine or frusemide. Neither peak nor trough aminoglycoside levels, nor duration of jaundice or level of bilirubin varied between groups. However, SNHL was more likely if peak bilirubin levels coexisted with netilmicin use (odds ratio (95% confidence interval) 14.2 (1.8 to 113.6)) or if acidosis occurred when bilirubin levels were over 200 micromol/l (OR 8.0 (0.9 to 71.6). Frusemide use in the face of high serum creatinine levels (OR 8.9 (1.1 to 74.5)) or netilmicin treatment (OR 5.0 (0.99 to 24.8)) was also associated with SNHL. At 12 months of age, seven of 15 children with SNHL had evidence of cerebral palsy compared with two of 30 controls (OR 12.3 (2.1 to 71)). CONCLUSIONS: Preterm children with SNHL required more intensive care in the perinatal period and developed more neurological complications than controls. Among very preterm babies, the coexistence of risk factors for hearing loss may be more important than the individual factors themselves. PMID- 10685989 TI - Severe retinopathy of prematurity and its association with different rates of survival in infants of less than 1251 g birth weight. AB - BACKGROUND: There is controversy over whether improved survival of preterm infants has resulted in a higher incidence of severe (grade 3 or greater) retinopathy of prematurity (ROP). AIM: To compare survival rates and rates of > or = stage 3 ROP-that is, with a high risk of sequelae-in preterm infants in five English cities where, anecdotally, the incidence of ROP is reported to show considerable variation. METHODS: All infants of birth weight < 1500 g and or gestational age < 32 weeks, born in 1994 in one of the cities or transferred in within 48 hours, were studied. The populations were adjusted for case mix variation using CRIB (clinical risk index for babies, a disease severity scoring system). The incidence of severe ROP, the actual death rate, and that adjusted for disease severity were determined. RESULTS: The rate of severe ROP per 1000 births was higher in city 1 than in all the other cities. This increase in comparison with city 2 and city 4 was significant (city 1, 167 (95% confidence interval (CI) 96 to 260); city 2, 24 (6 to 59); city 4, 16 (1 to 84)). A significant difference was not seen between city 1 and cities 3 (23 (1 to 120)) and 5 (74 (21 to 79)). The relative risk of developing severe ROP in city 1 compared with all the other cities was 5.5 (2.5 to 11.9). The actual death rate per 1000 births in city 1 was significantly lower than that predicted by modelling death against CRIB score (city 1: actual 270; predicted 385 (95% CI 339 to 431)). In contrast, the other cities had actual death rates as predicted, or worse than predicted, by CRIB. INTERPRETATION: A significantly higher incidence of severe ROP was identified in one of the five cities studied. Variation in survival rates among high risk infants may explain this observation. PMID- 10685990 TI - Haematocrit and red blood cell transport in preterm infants: an observational study. AB - AIMS: To test whether cardiac output acts as a compensatory response to changes in haematocrit. METHODS: A cohort of 38 preterm infants (27-31 weeks' gestation) was studied with repeated Doppler measurements of left ventricular output during the 1st month of life. Red blood cell transport was calculated when the duct was closed. RESULTS: Multiple regression analysis showed that left ventricular output correlated negatively with haematocrit when the duct was closed (n = 84) and when it was open (n = 59). The influence of an increase of 10% in haematocrit absolute value on mean (SD) left ventricular output was estimated at -55 (11) ml/kg/min. Mean (SD) red blood cell transport was 132 (30) ml/kg/min with a mean (SD) intra individual variability of 20% (8.8%). Red blood cell transport was increased more frequently by left ventricular output than by haematocrit. Haematocrit and left ventricular output but not red blood cell transport were dependent on postnatal age. CONCLUSION: These results suggest that in preterm infants cardiac output adaptation is effective in attenuating the effects of red blood cell mass variations on systemic oxygen carrying capacity. PMID- 10685991 TI - Measurement of interleukin 10 in bronchoalveolar lavage from preterm ventilated infants. AB - BACKGROUND: Interleukin 10 (IL-10) is a cytokine that downregulates inflammation, in part by reducing the production of the proinflammatory cytokines IL-1beta and IL-8. It has been suggested that an inability to produce IL-10 might predispose preterm infants to develop chronic lung disease. AIM: To measure IL-10, IL-1beta, and IL-8 in bronchoalveolar lavage fluid from ventilated preterm infants in a prospective cohort study. PATIENTS: 17 consecutive newborn infants < or = 29 weeks' gestational age (median, 25; 9 boys) who were ventilated from birth underwent daily bronchoalveolar lavage sampling. RESULTS: 102 samples were collected, of which 57 contained IL-10 in amounts that were comparable with those found previously in ventilated term infants with respiratory failure. Chronic lung disease developed in 9 of the 11 survivors and all 9 infants had produced IL 10. IL-1beta and IL-8 were detected in nearly all samples and were raised throughout the course of sample collection. CONCLUSION: IL-10 is readily detectable in early bronchoalveolar lavage samples from ventilated preterm infants, although it remains unclear whether this cytokine has any influence on the development of chronic lung disease. PMID- 10685992 TI - A randomised control study of partial liquid ventilation after airway lavage with exogenous surfactant in a meconium aspiration syndrome animal model. AB - AIMS: To test the hypothesis that lavage with exogenous surfactant before partial liquid ventilation in meconium aspiration syndrome (MAS) would improve debris removal, and therefore the effectiveness of partial liquid ventilation. METHODS: 12 newborn piglets were randomised into 4 groups, partial liquid ventilation or gas ventilation, with and without surfactant lavage. Physiological and blood gas data were compared between groups by analysis of variance. RESULTS: Arterial oxygen pressure (PaO(2)) was improved in the group treated with surfactant lavage when compared with the group not receiving surfactant. PaO(2) in the group receiving surfactant lavage followed by partial liquid ventilation was further improved when compared with the group treated with surfactant lavage followed by gas ventilation and the group receiving partial liquid ventilation alone. CONCLUSION: The effectiveness of partial liquid ventilation in MAS might be enhanced by pretreatment with exogenous surfactant bronchial lavage. PMID- 10685993 TI - Type 2 Gaucher disease: the collodion baby phenotype revisited. AB - The association of Gaucher disease, the inherited deficiency of lysosomal glucocerebrosidase (EC 3.2.1.45), and congenital ichthyosis was first noted a decade ago. Subsequently, a null allele type 2 Gaucher mouse was generated that also exhibited ichthyotic skin, confirming that the skin disorder and enzyme deficiency were directly related. This paper details the clinical and molecular characterisation of 6 cases of type 2 Gaucher disease presenting with the collodion baby phenotype. The identified mutant glucocerebrosidase alleles include two novel mutations (S196P and R131L) and two rare point mutations (R120W and R257Q), as well as alleles resulting from recombination with the nearby glucocerebrosidase pseudogene. There is significant genotypic heterogeneity in this rare subset of patients with type 2 Gaucher disease. Gaucher disease should be considered in the differential diagnosis of congenital ichthyosis in the newborn period. PMID- 10685994 TI - Proximity to maternity services and stillbirth risk. AB - A study of all 77 995 live births and 1234 stillbirths to mothers living in West Cumbria from 1950 to 1989 found no significant increase in stillbirth risk with distance of mother's residence from the first or second nearest maternity services, after allowing for year of birth, father's social class, and birth order. PMID- 10685995 TI - Dr Edward Rigby of Norwich (1747-1821) and antepartum haemorrhage. PMID- 10685996 TI - Apoptosis in transfusion medicine: of death and dying--is that all there is? PMID- 10685997 TI - Blocking T-cell costimulation in transplantation: opportunities and challenges. PMID- 10685998 TI - Committee report. Nucleic acid amplification testing of blood donors for transfusion-transmitted infectious diseases: Report of the Interorganizational Task Force on Nucleic Acid Amplification Testing of Blood Donors. PMID- 10685999 TI - White cell apoptosis in platelet concentrates. AB - BACKGROUND: The aim of the present study was the evaluation of the apoptosis in residual white cells (WBCs) contained in platelet concentrates (PCs) and of the relationship of this apoptosis with the concentration of inflammatory cytokines in the medium and with platelet activation. STUDY DESIGN AND METHODS: Three independent methods were used to evaluated apoptosis in WBCs present in 9 PCs, either from single donors by apheresis (SD-PCs) or from pooled buffy coats (BC PCs). All PCs were divided in two parts, one of which was irradiated. PCs were stored up to 4 days at room temperature, and samples were withdrawn daily for analysis of apoptosis, of platelet activation (surface and soluble CD62P), and of cytokine concentration (interleukin [IL]-1alpha, IL-1beta, IL-6, IL-8, and tumor necrosis factor alpha). RESULTS: Apoptosis was found to occur with storage in both irradiated and nonirradiated units. Platelet activation increased with storage time and was higher in BC-PCs. The amount of released cytokines was rather variable among PC units. Only IL-8 was consistently found to increase with storage time. CONCLUSIONS: Apoptosis of residual WBCs occurred in PC units as a function of storage time. The amount and the time course of apoptosis seem to correlate with IL-8 release rather than with platelet activation or with the occurrence of febrile nonhemolytic transfusion reactions. PMID- 10686000 TI - Nuclear matrix protein is released from apoptotic white cells during cold (1-6 degrees C) storage of concentrated red cell units and might induce antibody response in multiply transfused patients. AB - BACKGROUND: A previous study showed that white cells in blood units undergo apoptosis during storage. STUDY DESIGN AND METHODS: The present study attempts to show the release of nuclear matrix protein (NMP) in the supernatants of red cell units and to determine whether antibodies against nuclear components may be present in multiply transfused patients; the methods employed were enzyme-linked immunosorbent assay, flow cytometry, microscopy, immunoblotting, immunofluorescence, and confocal laser-scanning microscopy. RESULTS: NMP is released from white cells in the supernatant of packed red cell units upon cold storage (1-6 degrees C). The concentration of NMP correlates well with the degree of apoptosis, as analyzed by flow cytometry, nuclear dye staining, and DNA gel electrophoresis. Immunofluorescence also shows that white cells undergoing apoptosis (pre-G(1) peak, as seen by propidium iodide staining and flow cytometry) have an NMP content lower than control cells, which confirms an actual release of NMP. Moreover, immunoblotting analysis and immunofluorescent staining showed that, in 4 of 38 multiply transfused patients, autoantibodies against NMPs were present without any clinical or laboratory sign of autoimmune disease. One of the sera, recognizing a 64-kDa NMP, immunostained nuclear dots that were identified as coiled bodies because of their colocalization with p 80 coilin. CONCLUSION: NMP is released in the supernatant of red cell units. The results obtained from patients suggest that nuclear proteins released during apoptosis, once transfused, may induce an immune response in multiply transfused patients. PMID- 10686001 TI - High platelet contamination in progenitor cell concentrates results in significantly lower CD34+ yield after immunoselection. AB - BACKGROUND: Selection of CD34+ cells by specific immunoselection leads to a significant loss of those cells. The factors influencing the yield and purity are not well identified. The results of CD34+ selection from peripheral blood progenitor cells (PBPCs) with high and low platelet contamination that are harvested with two different cell separators are reported. STUDY DESIGN AND METHODS: A progenitor cell concentrator (Ceprate SC, CellPro) was used to select CD34+ cells from 41 PBPC concentrates from 23 consecutive patients with relapsed non-Hodgkin's lymphoma (n = 3), breast cancer (n = 17), and multiple myeloma (n = 3). PBPC collection was performed by using two cell separators (CS3000 Plus, Fenwal: Group A, n = 11; and Spectra, COBE: Group B, n = 9). To reduce platelet contamination in the Spectra PBPC concentrates, an additional low-speed centrifugation was performed before CD34+ cell selection (Group C, n = 3). Leukapheresis components were stored overnight at 4 degrees C and combined with the next day's collection before the CD34+ selection procedure in 19 patients. RESULTS: A median of 1.5 leukapheresis procedures per patient were performed. Pooled PBPC concentrates showed no statistical difference in median numbers of white cells and CD34+ cells in Groups A and B: 3.2 (0.8-9.2) versus 4.4 (1.6-8. 3) x 10(10) white cells per kg and 15.0 (4.7-24.0) versus 12.0 (5. 6-34.0) x 10(6) CD34+ cells per kg. Platelet contamination was significantly higher in Group B: 0.67 (0.15-2.4) versus 2.3 (0.5-7. 1) x 10(11) (p = 0.0273). After the selection process, there was a significantly greater loss of CD34+ cells in Group B than in Group A: 39.1 versus 63.2 percent (p = 0.0070), with a median purity of 78. 0 percent versus 81.0 percent. An additional low-speed centrifugation before CD34+ cell selection seemed to reduce CD34+ cell loss in Group C with 16.9, 31.9, and 37.5 percent, respectively. CONCLUSION: CD34+ cell selection from PBPC concentrates resulted in an increased loss of CD34+ cells in concentrates with a higher platelet content. To improve CD34+ yield, PBPC concentrates with an initially low platelet contamination should be used, or additional low-speed centrifugation should be performed. PMID- 10686002 TI - Apheresis-induced platelet activation:comparison of three types of cell separators. AB - BACKGROUND: Platelet-harvesting technology differs in various cell separators. Alteration in shear stress and biocompatibility of surfaces may give variable platelet activation and thereby affect the quality of the component. STUDY DESIGN AND METHODS: Four groups (n = 10) of single-needle apheresis procedures using three cell separators, were compared: 1) Spectra LRS, 90-minute harvesting time; 2) MCS+, 90-minute harvest; 3) Amicus, 90-minute; and 4) Amicus, 45-minute. Whole blood samples were collected from the donors as were samples from the final components at intervals during the first 4 hours after cessation of the apheresis. Platelet activation status and platelet activation capacity after agonist stimulation were assessed by flow cytometry. RESULTS: No activated platelets were found in preapheresis and postapheresis samples from the donors. The platelets in the components from the Amicus (90-min) were significantly more activated than those in the other groups of components: that is, there was increased size of platelet aggregates, increased fraction of microparticles, increased degranulation, increased fibrinogen receptor activation, and decreased von Willebrand factor receptor expression. Moreover, the response of these platelets to agonist stimulation was reduced for all activation variables. CONCLUSIONS: After 90 minutes' processing time, platelets obtained with the Amicus cell separator were significantly more activated than platelets harvested with the Spectra and the MCS+. PMID- 10686003 TI - Pressure cycling technology: a novel approach to virus inactivation in plasma. AB - BACKGROUND: Hydrostatic-pressure virus inactivation is a novel approach to the inactivation of pathogens in plasma and blood-derived components, that retains the therapeutic properties of these products. STUDY DESIGN AND METHODS: A custom built apparatus was used to pressurize human plasma samples spiked with lambda phage. Phage titer and plasma protein activities were monitored after pressure treatment. RESULTS: Pressure-mediated inactivation of lambda phage was found to be an effective means for virus inactivation, particularly when performed at near zero (0 degrees C) temperatures, rather than at temperatures above 20 degrees C and below -40 degrees C. The efficiency of inactivation was improved by an increase in applied pressure and repeated cycling from atmospheric to high pressure. In contrast, activities of plasma proteins alkaline phosphatase and total amylase did not vary with temperature and remained within 29 percent and 6 percent, respectively, of starting values after the same pressure treatments. By combining cycling, near-zero temperatures, and high pressure, phage titers in serum were reduced approximately 6 log after 10 to 20 minutes of treatment. Activities of plasma proteins IgG, IgM, and factor X were at 104 percent, 89 percent, and 80 percent, respectively, of starting values after 20 minutes of the same temperature and pressure treatment. CONCLUSION: High-pressure procedures may be useful for the inactivation of viruses in blood and other protein-containing components. PMID- 10686004 TI - A fully automated blood typing system for hospital transfusion services. ABS2000 Study Group. AB - BACKGROUND: The results of routine blood bank testing by a fully automated blood typing system (ABS2000) were compared with those obtained by standard manual methods in six hospital transfusion services. STUDY DESIGN AND METHODS: The ABS2000 system uses microtiter plates for determining ABO and D types, solid phase red cell adherence (SPRCA) assays for antibody detection, and modified SPRCA plates for IgG crossmatches. The transfusion services used their standard manual test tube methods. RESULTS: Of 3779 donors' samples tested for ABO types (red cell typings only), 3.0 percent could not be interpreted by the ABS2000 system's neural network, because of clots, hemolysis, or lipemic samples. The results for ABO types were concordant for 99.8 percent of the remaining samples. Of 3779 donors' samples tested for D types, the results were concordant for 98.7 percent. Of 7580 patients' samples tested for ABO types (red cell and plasma typings), 5.8 percent could not be interpreted by the ABS2000 system. There was 100-percent concordance of ABO typing results for the remaining 7140 samples. There was 99. 7-percent concordance of results for patients' D types. The results of 96.7 percent of antibody detection tests and 98.8 percent of crossmatches were concordant. Neither method failed to detect a serologically incompatible crossmatch that was associated with a specific, clinically significant alloantibody. The ABS2000 system performed 45 confirmatory donor ABO and D types in 115 minutes, 22 antibody detection tests in 116 minutes, 16 patients' ABO/D types in 149 minutes, and 40 crossmatches in 140 minutes. CONCLUSION: The ABS2000 blood typing system automates routine blood bank tests with accuracy comparable to that of hospital transfusion services' standard manual methods. PMID- 10686005 TI - Molecular basis of Cromer blood group antigens. AB - BACKGROUND: The Cromer blood group system consists of 10 antigens located on decay-accelerating factor (DAF). Previous molecular genetic analysis has determined the basis for four of these antigens. The present study was undertaken to identify the mutations that determine the remaining antigens. STUDY DESIGN AND METHOD: Existing or new data were used to localize each Cromer system antigen to a specific short consensus repeat (SCR) domain of DAF. The exon encoding that SCR domain was amplified by using the polymerase chain reaction (PCR) on genomic DNA obtained from individuals of that Cromer phenotype, and the DNA product was subjected to DNA sequence analysis. RESULTS: The Tc(a)/Tc(c) polymorphism is due to an R18P amino acid substitution in SCR1 of DAF. The Es(a+)/Es(a-) polymorphism is due to an I46N mutation in SCR1 of DAF. The WES(b)/WES(a) polymorphism is due to an L48R mutation in SCR1 of DAF. The UMC+/UMC- polymorphism is due to a T216M substitution in SCR4 of DAF. CONCLUSIONS: With information from previous reports and the findings of this study, the molecular genetic basis of all known alleles of the Cromer blood group system has been elucidated. Single amino acid substitutions are responsible for 9 of the 10 antigens (all except the multiple epitope antigen IFC). PMID- 10686006 TI - Expression of Rh30 and Rh-related glycoproteins during erythroid differentiation in a two-phase liquid culture system. AB - BACKGROUND: To gain insight into the formation of the Rh complex during erythroid differentiation, the ways in which Rh30 and Rh-related glycoproteins, especially Rh50, were produced in a modified two-phase liquid culture system were studied. STUDY DESIGN AND METHODS: A mononuclear cell fraction from fresh peripheral blood was first cultured in a medium supplemented with conditioned medium collected from the culture of a bladder carcinoma cell line (5637) for 7 days. Nonadherent cells were then collected for culture in a secondary medium containing 2 U per mL of erythropoietin to initiate erythroid differentiation. The expression of Rh30 and Rh50 during secondary culture (16 days) was monitored by flow cytometry. RESULTS: D+ cells appeared after Day 4 and increased to 70 percent by Day 8. On Day 12, 90 percent of the total cells became D+ and remained so until the end of the culture. A similar expression profile was obtained for Rh50. As determined from mean fluorescence intensities recorded in flow cytometry, the number of both D and Rh50 antigenic sites per cell increased as the differentiation progressed. Rh-related glycoprotein, CD47, had expression patterns significantly different from those of Rh30 and Rh50. In addition, the cultured cells produced partially glycosylated protein (approx. 32 kDa) in Rh50. CONCLUSION: Expressions of Rh30 and Rh50 occur simultaneously during erythroid differentiation, and both proteins are most actively synthesized at the last stage of the differentiation. In contrast, CD47 may be involved in expression of Rh30 in a different manner from Rh50. The two-phase liquid culture system will be an excellent model for studying the interaction among the components of the Rh complex during protein synthesis and complex assembly on the cell membrane. PMID- 10686007 TI - Neutrophil antigen 5b is carried by a protein, migrating from 70 to 95 kDa, and may be involved in neonatal alloimmune neutropenia. AB - BACKGROUND: Neutrophil antigen 5b has been described as involved in transfusion reactions and not in neonatal alloimmune neutropenia. CASE REPORT: Anti-5b was found in the serum of a mother of a persistently neutropenic newborn, who had several bacterial infections. The neutropenia responded to treatment with recombinant human granulocyte-colony-stimulating factor. Immunoprecipitation experiments performed with this and three other 5b antisera identified a protein, migrating from 70 to 95 kDa, as carrier of 5b. The observed pattern of migration may point to heavy glycosylation of this protein. RESULTS: Six 5b-negative donors were identified among 54 screened white donors, for a 5b gene frequency of 0.66. CONCLUSION: Alloimmunization to 5b in pregnancy is rare. In the patients with neonatal neutropenia analyzed in the last decade, this was the first case discovered. PMID- 10686008 TI - Comparison of flow cytometric assays with isotopic assays of (51)chromium-labeled cells for estimation of red cell clearance or survival in vivo. AB - BACKGROUND: A comparison was made between flow cytometric and conventional radioisotopic assays in the determination of the clearance or survival of small volumes of (51)chromium-labeled D+ red cells after injection into volunteers. STUDY DESIGN AND METHODS: Four clearance studies were performed using 4 mL of autologous D+ cells coated with anti-D at two concentrations (5 or 10 microg anti D/mL red cells) transfused to two subjects at separate times. Five survival studies were carried out using 5 mL of frozen-thawed D+ cells transfused to five D- subjects with no detectable anti-D. Sequential blood samples were taken for gamma counting and flow cytometry. Several methods were used to stain the transfused red cells, and the data were analyzed by using three flow cytometers. RESULTS: The determination of red cell clearance or survival by radioactivity measurements gave results consistent with published data. However, none of the flow cytometric assays exhibited the necessary sensitivity or accuracy in quantitation of the rare events to provide reliable data for the calculation of the initial clearance rate, the red cell half-life, or the mean cell lifespan, although rough estimates of red cell clearance were obtained in some subjects. This inability to accurately enumerate rare fluorescence-labeled cells was due mainly to the presence of "background" events, which were a considerable problem in some samples, when the coating level of anti-D was less than 3000 molecules of IgG per cell. CONCLUSION: Flow cytometry may enable the crude estimation of the percentage of small volumes (<5 mL) of transfused D+ red cells, but in this study it was found that this method was not sufficiently accurate to determine the initial clearance rate, red cell half-life, or mean cell lifespan. If the proportion of transfused cells in the recipient is about 0.2 percent or less, the use of radioisotopes for labeling cells for quantitative in vivo red cell clearance or survival data should remain the method of choice. PMID- 10686009 TI - DNA analysis in a paternity case involving a triploid fetus. AB - BACKGROUND: A parentage testing laboratory was asked to perform testing in a case of sexual assault that resulted in the conception of a child. Samples submitted to the laboratory included blood from the mother, the alleged father, and the fetus. CASE REPORT: DNA typing was used to determine if the suspect in this sexual assault was the father of the expected child. DNA extracted from these samples was subjected to both restriction fragment length polymorphism and polymerase chain reaction/short-tandem repeat analysis at a total of 13 genetic loci. Examination of DNA profiles for selected markers suggested that the fetus was triploid. Triploidy was confirmed through the use of fluorescent in situ hybridization of chromosomes, employing three chromosome-specific alpha satellite probes and fetal trophoblast nuclei. Statistical interpretation of the test results required identifying a method for calculation that would consider two transmitted paternal genes. Attempts to modify the standard method of calculating a paternity index were unsuccessful, because it was not possible to distinguish between dispermy and diandry as the mechanism of conception. Therefore, the likelihood ratio was calculated as the reciprocal of the random men not excluded value or the proportion of the population that possesses all of the paternal markers observed in the triploid fetus. CONCLUSION: Calculation of a likelihood ratio employing the exclusionary power of a collection of DNA markers appears to be the only method suitable for assigning weight to the significance of DNA matches between an alleged father and a child who is triploid. PMID- 10686010 TI - Prevalence of the newly described human circovirus, TTV, in United States blood donors. AB - BACKGROUND: A novel nonenveloped single-stranded circular DNA virus (TTV) was recently identified. The prevalence of TTV in blood donors in the United States is, however, still unclear. STUDY DESIGN AND METHODS: Viral DNA was detected in US blood donors from five cities by using two sets of TTV primers: NG059/NG061/NG063 primers, which amplified the conserved region of strains 1 and 2, and T801/T935 primers, which amplified the 5' end region of the TTV sequence. A TTV antibody assay system was based on the detection of the truncated open reading frame (ORF)-1 (amino acids 1-411) from type 1b. The truncated ORF-1 was expressed as a fusion protein in Escherichia coli, and the fusion protein was used as the antigen in the antibody assay system. RESULTS: Viremia was detected in 21 (8. 4%) of 250 donors by use of NG059/NG061/NG063 primers and 104 (41. 6%) of 250 by use of T801/T935 primers. There was little correlation among the assays, which suggests the preferential detection of different strains with the different primers. TTV antibody was detected in 38 of 100 donors: 32 (84%) of 38 with concurrent TTV viremia and 6 (16%) of 38 without TTV viremia. TTV viremia and/or TTV antibody-positive samples were detected in 52 (52%) of 100 of US blood donors. CONCLUSION: Evidence of infection or exposure to TTV appears to be common among blood donors in United States. PMID- 10686011 TI - Passive transfer of HIV-1 antibodies and absence of HIV infection after the transfusion of HIV-1-seropositive red cells. PMID- 10686012 TI - Blood component recalls in the United States, 1998. PMID- 10686013 TI - Putting the cart before the horse. PMID- 10686014 TI - Detection of Rh23 in the partial D phenotype associated with the D(Va) category. PMID- 10686016 TI - Notice of duplicate publication PMID- 10686015 TI - Do intravenous immunoglobulin products manufactured from plasma collected in Italy place immunocompromised patients at risk of contracting human herpesvirus 8? PMID- 10686017 TI - A large percentage of the Spanish population under 30 years of age is not protected against hepatitis A. AB - A seroepidemiological study was conducted to assess the seroprevalence of hepatitis A (HAV) antibodies in the Spanish general population in 1992-93. A total of 2744 subjects (1337 men and 1437 women) in the 5-59 years age range were stratified by gender and age (5-12, 13-19, 20-29, 30-39, 40-49, 50-59 years). The presence of total anti-HAV antibodies was investigated using a commercial enzyme immunoassay. Fifty-five percent (95% CI: 53.5-57. 2%) of the subjects were positive for anti-HAV antibodies, the age-standardized anti-HAV prevalence being 65.4%. Prevalence of seropositive subjects increased with increasing age (chi(2) = 996, 17; P < 0.0001), being 11%, 25% and 54% for the 5-12, 13-19 and 20-29 age groups respectively. The results from this study showed a remarkable decline in seroprevalence rates among children, adolescents and young adults. The large number of susceptible subjects in these groups of the population has public health implications in a country with intermediate HAV prevalence. PMID- 10686018 TI - The NS5a gene of hepatitis C virus in patients treated with interferon-alpha. AB - Patients infected with hepatitis C virus (HCV) genotype 3 have a better response to interferon-alpha (IFN-alpha) therapy than those infected with genotype 1. There are extensive sequence differences between genotypes in the 3' half of the NS5a gene. An association between IFN-alpha response and the interferon sensitivity-determining region (ISDR) (amino acids 2209-2248) of HCV genotype 1b has been described [Enomoto et al. (1996) New England Journal of Medicine 334:771 776]. A prospective study was conducted to determine whether the derived NS5A amino acid sequence or quasi-species diversity could predict response to IFN alpha therapy. Serum samples were obtained before, during, and after treatment from 35 IFN-alpha-treated patients chronically infected with HCV (eight with type1b,13 with type1a, and 14 with type3a). Nucleotide sequences were determined, and amino acid sequences corresponding to residues 2178-2390 of the polyprotein were derived. Quasi-species complexity was analysed by amplification of the ISDR region (2270-2403), followed by single-stranded conformation polymorphism (SSCP). No amino acid sequence that could be used to predict response to treatment was found, and there was no selection of specific amino acid residues during treatment. A striking lack of variability was seen in HCV genotype 3a, but the small degree of variation could suggest an effect on response. SSCP showed that variation in the predominant NS5a sequence occurred in the presence and absence of therapeutically administered IFN-alpha. HCV quasi-species diversity pretreatment did not predict IFN-alpha treatment outcome. The conclusion of the study is that the amino acid sequence of NS5a cannot be used to predict the efficacy of treatment with IFN-alpha in HCV-infected patients in Scotland. No evidence was found to support the selection of IFN-alpha-resistant strains in the NS5a gene. PMID- 10686019 TI - Recombinant subunit ORF2.1 antigen and induction of antibody against immunodominant epitopes in the hepatitis E virus capsid protein. AB - A recombinant subunit antigen (ORF2.1), representing the carboxy-terminal 267 amino acids of the 660-amino-acid hepatitis E virus (HEV) capsid protein, was expressed in Escherichia coli and used for the immunisation of rats. Purified antigen formulated with either Aluminium Hydroxide Gel Adjuvant (Alum) or Titermax gave high and equivalent levels of antibody after three doses. Responses to two doses of 15, 75, or 150 microg antigen, formulated with Alum and given at 0 and 4 weeks, were also equivalent by 17 weeks after immunisation. Rats initially developed antibody to a wide range of linear epitopes in the ORF2.1 region, but by 27 weeks the predominant response detected by Western immunoblotting was restricted to the conformational epitope unique to ORF2.1 [Li et al. (1997) Journal of Medical Virology 52:289-300], a pattern that was also observed when comparing acute-phase patient serum samples with serum samples from convalescing patients. Antibody from immunised rats blocked the majority of patients' serum reactivity in enzyme-linked immunosorbent assay against both ORF2.1 (57-92% inhibition) and virus-like particles of HEV produced using the baculovirus system (74-97% inhibition). Together, these results suggest that the ORF2.1 subunit vaccine induces an antibody response against immunodominant, conformational epitopes in the viral capsid, which largely mimics that seen in convalescent patients, who are presumed to be immune to HEV infection. PMID- 10686020 TI - TT virus infection in patients with chronic hepatitis B or C: influence on clinical, histological and virological features. AB - Concomitant infection with TT virus and hepatitis B virus (HBV) or hepatitis C virus (HCV) is common. However, the effect of TTV infection on chronic hepatitis B or C is unknown. The prevalence of TTV infection, the effect of TTV infection on the clinical, histological and virological features of patients with chronic hepatitis B or C, and the influence of TTV infection on the HCV response to interferon alfa therapy were studied. A total of 100 asymptomatic hepatitis B surface antigen carriers, 220 patients with HBV-related chronic liver diseases, and 110 patients with chronic hepatitis C treated with interferon alfa (3 million units subcutaneously three times a week for 24 weeks) were enrolled. Serum HCV RNA and serum TTV DNA were detected by the polymerase chain reaction (PCR). Serum HBV DNA and serum HCV RNA level were quantified by branched DNA assays. Infection with TTV was detected in 21.5% of HBV carriers and 37% of HCV carriers. TTV infection had little effect on the clinicopathological course of chronic HBV infection. In chronic hepatitis C, clinical features, histological severity, serum HCV RNA levels, and the response to interferon alfa therapy did not differ between those with and without TTV infection. The loss of serum TTV DNA did not correlate with the biochemical response as did in the loss of serum HCV RNA. In conclusion, TTV infection is found frequently in patients with chronic hepatitis B or C in Taiwan; however, coinfection with TTV does not affect the clinicopathological course of chronic hepatitis B or C and the response to interferon alfa therapy. PMID- 10686021 TI - Prevalence and risk factor analysis of TTV infection in prostitutes. AB - TTV, a DNA virus, has been isolated from patients with non-A to non-E post transfusion hepatitis. In the past it was assumed that TTV was transmitted parenterally. It is unclear whether sexual contact leads to transmission of this virus. In this study, two sets of TTV-specific polymerase chain reaction primers were used to detect serum TTV DNA in 140 prostitutes and 136 controls. The prevalence of TTV DNA in prostitutes was significantly higher than in the control group (46/140 [32.9%] vs. 29/136 [21.3%]; P = 0.043). There was no significant difference in the prevalence of positive antibody to hepatitis A virus (anti-HAV) in either group (87.8% for prostitutes, 85.3% for controls). No particular risk factor was significantly associated with positive TTV DNA in prostitutes. In summary, TTV is highly prevalent in prostitutes. Transmission of TTV via sexual contact is not as efficient as transmission of hepatitis C and D viruses and GB virus-C hepatitis G virus. The high prevalence of TTV in controls indicates that there are diverse routes of transmission of this virus. PMID- 10686022 TI - Physical state and expression of human papillomavirus in laryngeal carcinoma and surrounding normal mucosa. AB - Epidemiologic and biomolecular evidence suggests that human papillomavirus (HPV) infection may be associated with the development of head and neck cancers. To clarify the role of HPV in larynx carcinoma, 25 patients were studied for the presence of viral DNA, possible virus integration into the cellular genome, and viral expression both in neoplastic tissues and in neighbouring normal mucosa. Twelve of 25 patients with neoplasia (48%) showed negative results for HPV sequences, and 13 (52%) showed positive results. Among the latter group of patients, seven were HPV-16 positive, five were HPV-6, and one was HPV-45. No multiple infections were detected. The physical status of the HPV genome was analysed by three methods: polymerase chain reaction (PCR), bidimensional agarose gel electrophoresis, and in situ hybridisation. Viral integration into the host genome occurred in 43% of cases of HPV-16 and in 20% of cases of HPV-6. Viral RNA expression was detected by reverse transcription-PCR only in HPV-16-positive tumours. The pattern of expression was consistent with an active role of HPV in cellular transformation. In conclusion, the present work suggests that HPV infection may be involved in some cases of laryngeal carcinoma. However, the transformation mechanisms might be different from those currently accepted for anogenital cancers. PMID- 10686023 TI - Seroresponses to human papillomavirus types 16, 18, 31, 33, and 45 virus-like particles in South African women with cervical cancer and cervical intraepithelial neoplasia. AB - The aim of the study was to determine the prevalence of antibodies to human papillomavirus (HPV) types 16, 18, 31, 33, and 45 in woman in Cape Town with cervical intraepithelial neoplasia (CIN) (n = 95), cervical cancer (n = 40), female blood donors (n = 95) and children (n = 110). The enzyme-linked immunosorbent assay (ELISA) made use of baculovirus synthesised HPV virus like particles (VLPs) as antigen. Antibodies to at least one HPV type were detected in sera from 75% of cancer patients, 71.6% of CIN patients, 44.2% of blood donors and 27.3% of children. Sera from 95 women with CIN were compared with age-matched female blood donors. There was a significant association of seropositivity to VLP 16 (P = 0.006) and VLP-45 (P = 0.008) with CIN compared with the blood donors. There was also a significant difference in the seropositivity of women with CIN to any of the five virus-like particle (VLP) types compared to the blood donors (P = 0.0002: OR = 3.2). Thirty-nine of sixty-nine (56.5%) women with CIN were found to be HPV-16 DNA positive. The average age of women in this group that were VLP-16 seropositive was 34 years and those found to be VLP-16 seronegative was 52 years of age. Antibodies to all five VLP types were detected in these populations, thus an ideal vaccine should induce protection from infection by a wide range of HPV types. PMID- 10686024 TI - Epstein-Barr virus-specific antibodies in Epstein-Barr virus-positive and negative gastric carcinoma cases in Japan. AB - We examined Epstein-Barr virus (EBV)-specific antibodies in serum samples from 64 and 59 patients with EBV-positive and -negative gastric carcinomas, respectively, and 73 healthy controls using immunofluorescence assays. EBV capsid antigen (VCA) IgG and EBV-determined nuclear antigen (EBNA) IgG were detected in all 196 subjects. The geometric mean titer (GMT) of VCA-IgG, but not EBNA-IgG, was higher in EBV-positive carcinoma cases than in EBV-negative carcinoma cases (P < 0.001). The seroprevalence rates of VCA-IgA and EBV early antigen (EA) IgG were higher in EBV-positive carcinoma cases than in EBV-negative carcinoma cases. Odds ratios (ORs) comparing seroprevalence rates between EBV-positive and -negative carcinoma cases were 3.4 (95% confidence interval [CI] = 1.3-8.8) and 6.6 (95% CI = 2.7 16.3) for VCA-IgA and EA-IgG, respectively. These results suggest that EBV reactivation occurs in vivo, since more than 90% of Japanese are infected with EBV in early childhood. The GMT of VCA-IgG in EBV-negative carcinoma cases was higher than that of healthy controls (P = 0.028). The seroprevalence rates of EA IgG were greater in EBV-negative carcinoma cases than in healthy controls (OR = 4.9, 95% CI = 1.2-19. 7). VCA-IgA was the only antibody that showed a significantly high seroprevalence and GMT in EBV-positive carcinoma cases, but not in EBV-negative carcinoma cases. Thus, VCA-IgA can be a marker of immune response to EBV in EBV-positive carcinoma cases. Our findings support the hypothesis that if EBV is involved in the development of EBV-positive gastric carcinoma, the EBV reactivation occurs in vivo. PMID- 10686025 TI - Augmentation of leukocyte infiltration in murine tumors expressing B-cell derived but not nasopharyngeal carcinoma derived EBV membrane protein LMP1. AB - The Epstein-Barr virus (EBV) encoded latent membrane protein of B cell origin, B LMP1 (B95-8 prototype) and nasopharyngeal carcinoma (NPC) derived C-LMP1 (CAO prototype) were transfected individually in S6C adenocarcinoma cells of ACA (H 2f) origin. We have shown previously that inoculation of B-LMP1 expressing S6C cells led to tumor rejection in pre-immunized, immunocompetent syngeneic ACA mice, whereas the C-LMP1 transfectants were not immunogenic. Furthermore, B-LMP1 but not C-LMP1 expressing S6C cells grew with necrosis and extensive skin damage in non-immunized mice. A study was carried out to determine whether the in vivo growth pattern of S6C cells expressing two different LMP1 isolates could be correlated to any immunomodulatory mechanism. An increased infiltration of CD45+ leukocytes was found in B-LMP1 expressing S6C tumors originating in non immunized, syngeneic ACA mice. The C-LMP1 expressors, vector transfectants and untransfected parental tumors had significantly lower number of infiltrating leukocytes. The immunoaccessory molecules ICAM-1, B7-1 and MHC Class I and II expression was unaltered in both B- and C-LMP1 transfectants. The data suggest that B-LMP1 but not C-LMP1 induce anti-tumor immune response. PMID- 10686026 TI - Infection of five human liver cell lines by dengue-2 virus. AB - Elevated serum transaminase levels of dengue patients indicate the possible impact of dengue virus infection on liver function. To elucidate the action of dengue virus infection in liver cells, an in vitro cell line system was established that mimicked the liver status of diverse clinical patients. Briefly, four hepatoma cell lines (HA22T, Huh7, Hep3B, and PLC) and one nonmalignant hepatocyte cell line (Chang liver) were included, representing various levels of tumorigenicity and differentiation. Our data showed that in these five cell lines, dengue-2 virus attached to each cell type equally well; however, this virus had higher replication rates and levels of virion production in differentiated Huh7, PLC, Hep3B, and Chang liver cells. Likewise, a lower replication rate was observed in the de-differentiated HA22T cells. Differentiation-related factors seem to play an important role in dengue virus replication. Further study showed that sodium butyrate (NaB, a differentiation inducer) treatment enhanced dengue virus replication in HA22T cells. Moreover, we found that the severity of morphologic aberration and the increase in aspartate aminotransferase (AST) levels correlated with the virus replication rate in the four infected hepatoma cells. In conclusion, we showed that dengue virus can infect diverse liver cells with differing replication efficiency, which causes cytopathic effects (CPEs) of diverse severity. Among the CPEs, the increased AST levels correlated with the clinical results from 24 dengue fever patients, who showed increased AST levels at the onset of fever. In summary, we find that dengue-2 virus replicates actively and causes severe CPEs in differentiated hepatoma cells. Factors related to differentiation as well as tumorigenicity seem to play critical roles, though the mechanisms of action remain unclear. PMID- 10686027 TI - Prospective study of the duration and magnitude of viraemia in children hospitalised during the 1996-1997 dengue-2 outbreak in French Polynesia. AB - The magnitude and duration of viraemia in children admitted to the hospital with dengue was studied during a dengue 2 outbreak in French Polynesia in 1996-1997. Forty-nine patients from whom at least 3 plasma samples were available were included in the study. Based on analysis of IgG-ELISA and haemagglutination inhibition assay, 21 of these were primary and 28 were secondary infections. According to World Health Organization criteria, 42 were dengue fever and 7 were dengue haemorrhagic fever. Virus was detectable by reverse transcription-PCR in all patients for at least the first 3 days of the onset of fever, but was never detected after the 6th day (mean duration = 4.4 days). Plasma virus titers ranged from 1.7-5.6 Log(10) TCID(50)/ml. A significant difference was not observed in the magnitude and duration of viraemia in patients with primary versus secondary infections. The severity of the illness, however, was correlated with both criteria. PMID- 10686028 TI - A new enzyme immunoassay for the detection of enteroviruses in faecal specimens. AB - A new enzyme immunoassay (EIA) for direct detection of enteroviruses based on a group-specific monoclonal antibody was evaluated using stool samples from patients with suspected enteroviral infection. The EIA was compared with polymerase chain reaction (PCR) and virus isolation in cell culture. Of 204 samples tested, 20 were positive by EIA, 34 by PCR, and 18 by cell culture. Compared with PCR, the most sensitive method, the sensitivity of EIA was 58% (20/34); the sensitivity of cell culture isolation was 52% (18/34). The results of both assays correlated in only 60% of cases. The combination of EIA and cell culture isolation detected 76% of PCR-positive stool samples. Enterovirus EIA provides results within 3-4 hr and requires only standard EIA equipment. It represents a rapid, reliable, and cost-effective diagnostic tool for enterovirus diagnosis from faecal samples. Negative results must be confirmed by other techniques, such as PCR or virus isolation in cell culture. PMID- 10686029 TI - A neutralizing recombinant human antibody Fab fragment against Puumala hantavirus. AB - A combinatorial human antibody Fab pComb3H library, generated from splenic lymphocytes of a Puumala hantavirus (PUUV) immune individual, was selected against PUUV using the phage display technique. Panning was carried out with antigens immobilized by MAbs directed to the two PUUV envelope glycoproteins G1 and G2. Thirteen Fabs, with reactivity directed to PUUV and specifically the G2 protein, as assessed by immunofluorescence and ELISA respectively, were isolated in crude preparations. By a focus reduction neutralization test (FRNT), four of the 13 crude Fab preparations exhibited type-specific neutralization of PUUV (strain Sotkamo) with 44-54% reduction in the number of foci. After affinity purification, the four Fab clones exhibited 50% focus reduction of PUUV at concentrations below 2 microg/ml. Sequencing of the heavy and light chain complementarity determining regions (CDR) 1-3 showed that the four selected clones were identical within the antibody binding regions. In inhibition tests with the PUUV G2-specific MAbs, 4G2 and 1C9, a new epitope important for neutralization, designated as G2-a3, was defined. This epitope, overlapping partially the neutralizing epitope recognized by the human MAb 1C9, seems to be unique for the PUUV serotype since none of the Fab clones neutralized any of the other hantaviruses tested. PMID- 10686030 TI - Quantitative analysis of cytomegalovirus load using a real-time PCR assay. AB - A novel real-time PCR assay system was developed to quantify the cytomegalovirus (CMV) genome load. The real-time PCR assay could detect from 6 to over 10(6) copies of CMV-DNA with a wide linear range. The virus load of immunocompromised patients with symptomatic CMV infections was quantified and compared to that of asymptomatic ones. In symptomatic patients, all 17 peripheral blood leukocytes were positive for CMV DNA, and its mean value was 10(3.3) copies/10(6) cells. On the other hand, only 9 of 38 samples (24%) were positive in the asymptomatic patients, and its mean titer was lower (10(2.0) copies/10(6) cells) than that of the symptomatic group (P = 0.002). In plasma, the virus genome was detected in 13 out of 17 samples from symptomatic patients (76%), and its mean value was 10(4.0) copies/ml. In contrast, for the asymptomatic group, only one out of 36 samples were positive (3%). Finally, this system was used to monitor two patients with CMV infections serially. The CMV DNA copy number changed with their clinical symptoms and anti-CMV therapy, and virtually paralleled the result of the pp65 antigenemia assay in both cases. In one patient with the cord blood transplantation, however, the CMV DNA became positive faster than the antigenemia assay. These results indicate that this assay is sensitive and useful for estimating the CMV genome load not only in peripheral blood leukocytes but also in plasma. It can be very helpful for diagnosing CMV-related diseases and monitoring the virus load in patients with CMV infections. PMID- 10686031 TI - Diagnosis of Ebola haemorrhagic fever by RT-PCR in an epidemic setting. AB - This study reports the first field evaluation of a new diagnostic technique for Ebola virus disease with sensitivity and specificity. Ebola virus causes rare but fulminating outbreaks in Equatorial Africa. Rapid differentiation from other infections is critical for timely implementation of public health measures. Patients usually die before developing antibodies, necessitating rapid virus detection. A reverse transcriptase-polymerase chain reaction (RT-PCR) assay was developed, implemented and evaluated at Centre International de Recherches Medicales de Franceville (CIRMF) in Gabon, to detect Ebola viral RNA in peripheral blood mononuclear cells (PBMC). Twenty-six laboratory-confirmed patients during and 5 after the acute phase of Ebola haemorrhagic fever, 15 healthy controls and 20 febrile patients not infected with Ebola virus were studied. RT-PCR results were compared with ELISA antigen capture, and Ebola specific IgM and IgG antibody detection. Ebola virus RNA was amplified from 26/26 specimens from the acute phase, 3/5 during recovery, 0/20 febrile patients and 1/15 negative controls. Sensitivity of RT-PCR in identifying acute infection and early convalescence compared with antigen or IgM detection was 100% and 91% respectively, and specificity compared with antigen detection and IgM assay combined was 97%. Antigen capture detected only 83% of those identified by PCR, and IgM only 67%. Ebola virus RNA was detected in all 13 fatalities, only 5 of whom had IgM and none IgG. RT-PCR detected Ebola RNA in PBMC one to three weeks after disappearance of symptoms when antigen was undetectable. RT-PCR was the most sensitive method and able to detect virus from early acute disease throughout early recovery. PMID- 10686032 TI - Evaluation of the antibody specificities of human convalescent-phase sera against the attachment (G) protein of human respiratory syncytial virus: influence of strain variation and carbohydrate side chains. AB - The C-terminal third of the attachment protein (G) of several human respiratory syncytial virus isolates was obtained as either a glycosylated protease-resistant fragment of the purified protein or a nonglycosylated GST fusion protein expressed in bacteria. The reactivity of human convalescent-phase sera with both forms of the protein segment was evaluated in immunoblots. While all serum samples reacted with the mature intact protein of the different isolates, only certain samples reacted with the nonglycosylated C-terminal segment of some viral isolates. The number of human serum samples reacting with the glycosylated C terminal fragment was even more limited. These results highlight the heterogeneity of the human antibody response against epitopes located in the C terminal hypervariable region of the G molecule and the influence of carbohydrate side chains for expression of these epitopes. We also have analysed the specificities of human sera by competitive enzyme-linked immunosorbent assay with murine monoclonal antibodies (MAbs). Most human serum samples inhibited virus binding of MAbs that recognised conserved or group-specific epitopes of the G protein, while only a limited fraction of those samples inhibited binding of MAbs that recognised strain-specific epitopes. These results are discussed in terms of the antibody repertoire induced after human respiratory syncytial virus infection and the relevance of escape mechanisms to preexisting antibodies for the evolution of this virus. PMID- 10686033 TI - Expression of recombinant Norwalk-like virus capsid proteins using a bacterial system and the development of its immunologic detection. AB - The capsid protein of Norwalk-like virus (NLV) isolates NLV-36 (Mexico virus type, genogroup II [GII]), NLV-21 (Lordsdale virus type, GII), NLV-114 (untyped GII virus), and NLV-96-908 (KY89 virus type, GI) have been expressed in an Escherichia coli system. The expressed recombinant NLV capsid proteins, fused with maltose binding protein (MBP-rV) and thioredoxin (TRX-rV) in E. coli lysate, were analyzed using sodium dodecyl sulfate-polyacrylamide gel eletrophoresis. Rabbit IgG (R-IgG) in hyperimmune serum has been raised against MBP-rV-36 capsid protein and was purified before further study. Detection of TRX-rVs using an enzyme-linked immunosorbent assay (ELISA) showed that R-IgG had immunologic reactivity to GII as well as to the GI rV capsid proteins TRX-rV-36, TRX-rV-21, TRX-rV-114, and TRX-rV-96-908. Results of Western immunoblot (WB) analysis showed the same broad recognition of R-IgG when using the same samples. The results of the ELISA tests on serum samples obtained from patients involved in confirmed outbreaks of NLV proved that expressed NLV capsid proteins in E. coli can be detected by NLV-infected human serum. In addition, purified NLVs (LD virus types) derived from patients' stool could be detected using anti-NLV R-IgG, whereas normal R-IgG did not react when using WB. Our results strongly suggest that the immunologic detection of NLV antigens using anti-rV R-IgG is possible and seems a significant step toward simplification of an NLV detection test. PMID- 10686034 TI - Evidence for the diagnosis of pancreatic insufficiency. PMID- 10686035 TI - Long-acting beta(2)-agonists in childhood asthma: Don't change a winning team (yet) PMID- 10686036 TI - Evaluation of fecal pancreatic elastase-1 as a measure of pancreatic exocrine function in children with cystic fibrosis. AB - Pancreatic elastase-1 (EL-1) is a specific human protease synthesised by the acinar cells. It is stable, unaffected by exogenous pancreatic enzyme treatment, and correlates well with stimulated pancreatic function tests. We report our experience of EL-1 measurements in 142 patients from a large cystic fibrosis (CF) clinic. The median patient age was 7.7 years (range, 0.1-20.8 years), 93 were homozygous and 38 heterozygous for DeltaF508, and 11 had other or unidentified mutations. There were 85 non-CF control subjects. Seven were pancreatic sufficient (PS). The median (quartile 1-quartile 3) fecal EL-1 of the 135 pancreatic insufficient (PI) patients was 10 microg/g stool (2.5-33); of the 7 PS patients, 698 microg/g stool (400.5-824.5), and of the non-CF controls, 615 microg/g stool (420-773). Using the Mann-Whitney U test, there was a statistically significant difference for fecal EL-1 activity between the PS and PI patients (P = 0.0001) and the PI and control group (P < 0.0001), but not between the control and PS groups (P = 0.63). Median (quartile 1-quartile 3) fecal EL-1 in the pancreatic insufficient DeltaF508 homozygotes was 10 microg/g stool (2-33), and in the heterozygotes 12 microg/g stool (4-39) (not significant, P = 0.62). We now use fecal EL-1 as evidence of PI in screened CF infants (reliable over the age of 2 weeks); in older CF patients at diagnosis; for confirming the need for pancreatic enzymes in patients referred to the clinic already taking enzymes; for annual monitoring of PS patients to detect the onset of PI; and as supporting evidence when excluding the diagnosis of CF in patients attending the pediatric gastroenterology clinic. The low values in the first 2 weeks in some normal and premature infants, and the persisting normal values in PS infants, make the fecal EL-1 test unsuitable for neonatal CF screening. PMID- 10686037 TI - Pseudomonas aeruginosa cross-infection among patients with cystic fibrosis during a winter camp. AB - Twenty-seven patients with cystic fibrosis from our Danish Cystic Fibrosis Center went to a winter camp for 1 week in November of 1990. This study is based on 22 of these patients. Prior to attending camp, 17 out of 22 patients harbored Pseudomonas aeruginosa in their sputum, but 5 patients did not. After returning from camp, all 22 patients harbored P. aeruginosa in the sputum, including the 5 patients whose sputum was free of P. aeruginosa before they went. Epidemiological typing used pulsed-field gel electrophoresis of the P. aeruginosa isolates was performed. The typing results showed that the 5 cystic fibrosis patients who were free of P. aeruginosa in their sputum prior to the winter camp had acquired P. aeruginosa isolates identical to the P. aeruginosa strains isolated from the other 17 cystic fibrosis patients. This constitutes a cross-colonization rate of 100%, the highest rate ever detected among patients with cystic fibrosis. We conclude that separate holiday camps based on the infection status of the patients with cystic fibrosis are necessary to avoid cross-infection of patients not infected with P. aeruginosa. PMID- 10686038 TI - Bronchiectasis in Alaska Native children: causes and clinical courses. AB - Although bronchiectasis has become a rare condition in U.S. children, it is still commonly diagnosed in Alaska Native children in the Yukon Kuskokwim Delta. The prevalence of bronchiectasis has not decreased in persons born during the 1980s as compared with those born in the 1940s. We reviewed case histories of 46 children with bronchiectasis. We observed that recurrent pneumonia was the major preceding medical condition in 85% of patients. There was an association between the lobes affected by pneumonia and the lobes affected by bronchiectasis. Eight (17%) patients had surgical resection of involved lobes. We conclude that the continued high prevalence of bronchiectasis appears to be related to extremely high rates of infant and childhood pneumonia. Pediatr Pulmonol. 2000;29:182-187. Published 2000 Wiley-Liss, Inc. PMID- 10686039 TI - Effects of high-dose inhaled corticosteroids on bone metabolism in prepubertal children with asthma. AB - We studied the effect of inhaled corticosteroids on the increase in bone mineral content in prepubertal children with asthma. Forty-eight asthmatic, prepubertal children receiving either inhaled beclomethasone dipropionate or budesonide were evaluated. Nine children of similar age not receiving inhaled steroids served as controls. The average age of corticosteroid-treated children was 7.8 +/- 2.4 years, and of control children, 8.4 +/- 2.1 years (NS). The average dose of inhaled corticosteroids in the treated children was 0.67 +/- 0.48 mg/m(2)/day, and they were followed over a 9-20-month period. Total bone mineral content (TBMC) was measured at baseline and after 9-20 months. A derived value for 12 months' TBMC was calculated, assuming that changes in TBMC were linear with the passage of time. The change in TBMC over a 12-month period was 264 +/- 68 mg for the corticosteroid-treated children and 330 +/- 84 mg for control children (P < 0.025). In a multiple regression analysis in which adjustments were made for the effects of age, height, and weight, the change in TBMC in corticosteroid-treated children was inversely related to the inhaled steroid dose/m(2)/day (P = 0.016). The increase in the lumbar vertebral bone mineral density in control children was also significantly greater than in the corticosteroid-treated children (P < 0.025). We conclude that inhaled steroids, at an average dose of 0.67 mg/m(2)/day, when used in the treatment of asthma reduce the acquisition of bone mineral in prepubertal children. PMID- 10686040 TI - Comparative study using oral solutions of bambuterol once daily or terbutaline three times daily in 2-5-year-old children with asthma. Bambuterol Multicentre Study Group. AB - The aim of this study was to compare safety and efficacy of bambuterol hydrochloride (10 mg) oral solution administered once daily in the evening with terbutaline sulphate (0.075 mg/kg body weight) oral solution administered three times daily in 2-5-year-old children with asthma. There were two treatment groups: (2/3) of the patients received bambuterol and (1/3) received terbutaline. The study was double-blind, randomized, and of a parallel group design, and it lasted for 3 months after a 2-week run-in period. The primary objective was to evaluate safety (adverse events, and changes in blood pressure, pulse rate, hematology, and clinical chemistry parameters). Plasma concentrations of terbutaline and/or bambuterol were also measured. Evaluation of efficacy (diary card data) was a secondary objective. A total of 155 patients (range, 2-6 years; 3 patients were 6 years old at randomization) were treated with the study drugs; 104 patients received bambuterol and 51 patients received terbutaline. Both treatments showed a good safety profile with respect to clinical and laboratory tests, and they were generally well tolerated. Reported adverse events were mild to moderate. There were no statistically significant differences between treatment groups in any of the efficacy variables (diary variables: peak expiratory flow (PEF), asthma symptoms, restlessness, other reported symptoms, use of inhaled bronchodilators, and nighttime awakenings). For morning PEF, the mean increase from run-in to treatment was 16.9 L/min in the terbutaline group and 23.3 L/min in the bambuterol group. For evening PEF, the mean increase was 20.2 L/min in the terbutaline group and 20.6 L/min in the bambuterol group. In conclusion, once-daily bambuterol is as safe and effective as terbutaline given three times daily. The study also confirmed that bambuterol has a 24-hr duration of action, and therefore its once daily administration, makes it a preferred bronchodilator agent. Pediatr Pulmonol. 2000:29:194-201. PMID- 10686041 TI - Effects of inhaled nitric oxide and surfactant treatment on lung function and pulmonary hemodynamics in bronchoalveolar-lavage-induced respiratory failure. AB - Our aim was to study whether inhaled nitric oxide (iNO) moderates respiratory failure induced by bronchoalveolar lavage (BAL) without severe pulmonary hypertension. The following successive treatments, interrupted by 20-30-min rest periods, were given to piglets: iNO (20 ppm for 20 min), exogenous surfactant, iNO, Nomega-nitro-L-arginine methyl ester (L-NAME), and iNO. The controls inhaled NO first after L-NAME. Lung mechanics and hemodynamics were measured serially. The pulmonary to systemic arterial pressure ratio decreased during iNO and tended to increase after its discontinuation. In contrast, the iNO-induced decreases in severity of respiratory failure were not reversible during the rest periods. In a second experiment, iNO/placebo and surfactant containing (3)H-labeled dipalmitoyl phosphatidylcholine were given to rabbits. The surfactant aggregates and the surface activity from postmortem BAL, and extravascular lung water, were studied. Inhaled NO improved the surface activity and increased the large surfactant aggregates. There was no detectable decrease in extravascular lung water. The results suggest that a low dose of iNO has a beneficial effect on the gas exchange that is in part unrelated to its effect on the pulmonary vasculature. PMID- 10686042 TI - In vitro activity of minocycline against respiratory pathogens from patients with cystic fibrosis. AB - Our objective was to determine the in vitro activity of minocycline against isolates of Burkholderia cepacia (BC), Stenotrophomonas maltophilia (SM), and Pseudomonas aeruginosa (PA) cultured from the respiratory tract of patients with cystic fibrosis (CF). Cultures of BC, SM, and PA were isolated in a hospital bacteriology laboratory from the sputum or oropharyngeal cultures obtained from patients attending a Cystic Fibrosis Center, and were prospectively tested for in vitro sensitivity to minocycline by Kirby-Bauer disk diffusion. From January 1994 to July 1995, 116 cultures from 61 patients had at least one of the three pathogens; 9/61 (15%) patients had an isolate of BC, and 7/9 (78%) had an initial isolate sensitive to minocycline, of which 3 were sensitive only to minocycline; 2 cultures were resistant to all antibiotics. Four of 7 patients with BC were treated with minocycline; 3 patients developed resistant isolates 3-13 months after therapy. Five of 61 patients (8%) had an isolate of SM: 4/5 (80%) of these isolates were sensitive to minocycline, of which 1 was sensitive only to minocycline. Fifty-five of 61 patients (90%) had at least one PA isolate, with 112 morphotypes recovered from 90 cultures: 40/112 morphotypes (36%) were sensitive to minocycline, 65 (58%) were resistant, and 7 (6%) were intermediate in sensitivity. We conclude that the marked in vitro activity of minocycline against BC and SM isolated from patients with CF suggests that minocycline may have an adjunct role in the antimicrobial therapy of multidrug resistant, respiratory pathogens in CF. PMID- 10686043 TI - Interrupter technique versus plethysmography for measurement of respiratory resistance in children with asthma or cystic fibrosis. AB - The purpose of the present study was to compare measurements of respiratory system resistance by the interrupter method (Rrsint) with those of airway resistance by plethysmography (Raw) in nonobstructed children with asthma or cystic fibrosis (ratio of forced expiratory volume in 1 sec to vital capacity, FEV(1)/VC >/=80% with a forced expiratory flow rate between 25-75% of forced vital capacity, FEF(25-75) >/=75% of normal values) and in obstructed children with the same diseases (FEV(1)/VC <80% and/or FEF(25-75) <75% of normal values). Eighty-one children (47 asthmatics and 34 suffering from cystic fibrosis) aged 5 18 years (mean 11.2 +/- SD 3.4 years) were included in the study. For the overall group, we observed generally lower values for Raw (4.7 +/- 2. 8 cmH(2)O.L(-).s) than for Rrsint20 (extrapolation of the mouth pressure during occlusion to 40 ms after interruption) (5.6 +/- 1.7 cmH(2)O.L(-1).s) (P < 0.02), or for Rrsint40 (extrapolation of the mouth pressure during occlusion to 60 ms after interruption) (6.6 +/- 2.2 cmH(2)O.L(-1).s) (P < 0.001), but there was no difference between Rrsint20 and Raw in the obstructed subgroup. Moreover, we observed a correlation between the difference (Rrsint20 - Raw) expressed in percentage of predicted values and the degree of obstruction estimated by FEV(1)/VC (r = 0.56, P < 0.001). The differences between the specific resistances (sRrsint20 - sRaw, sRrsint40 - sRaw) were also correlated with the severity of the obstruction (r = 0.65, P < 0.001 and r = 0.57, P < 0.001, respectively). We observed also that the tendency to underestimate resistance by Rrsint in obstructed children was not the same in children with asthma and cystic fibrosis. We conclude that the tendency of Rrsint, as measured with our method, to underestimate airway obstruction appears to increase in proportion to the severity of the airway obstruction. PMID- 10686044 TI - Long-acting beta(2)-agonists in management of childhood asthma: A critical review of the literature. AB - This review assesses the evidence regarding the use of long-acting beta(2) agonists in the management of pediatric asthma. Thirty double-blind, randomized, controlled trials on the effects of formoterol and salmeterol on lung function in asthmatic children were identified. Single doses of inhaled salmeterol or formoterol cause prolonged bronchodilatation (>12 h) and extended bronchoprotection against exercise-induced bronchoconstriction in children, some children achieving full protection for more than 12 h. Heterogeneity in bronchoprotection has been observed, and individual dose-titration may be attempted. The onset of action of formoterol is comparable to salbutamol, while salmeterol has a slower onset of action. Partial tolerance develops when long acting beta(2)-agonists are used as regular treatment, including cross-tolerance to short-acting beta(2)-agonists. Regular treatment with salmeterol in children with or without corticosteroids provides statistically significant bronchodilatation, but the degree of improvement in lung function or bronchoprotection against exercise and nonspecific irritants is small with regular use. There is no evidence of anti-inflammatory effects from inhaled long acting beta(2)-agonists, which is reflected by unchanged or increased bronchial hyperreactivity and no reduction of exacerbation rates. The evidence does not support a recommendation for long-acting beta(2)-agonists as monotherapy, nor does it support their general use as regular add-on therapy. In conclusion, long acting beta(2)-agonists provide effective bronchodilatation and bronchoprotection when used as intermittent, single-dose treatment of asthma in children, but not when used as regular treatment. Future studies should examine the positioning of long-acting beta(2)-agonists as an "as needed" rescue medication instead of short acting beta(2)-agonists for pediatric asthma management. PMID- 10686045 TI - Pneumocystis carinii isolated from lung lavage fluid in an infant with cystic fibrosis. AB - Pneumocystis carinii (P. carinii) cysts were identified in bronchoalveolar lavage fluid from a 15-week-old child newly diagnosed with cystic fibrosis who presented with bronchitis, pneumonia, and weight loss. The child was not infected with human immunodeficiency virus (HIV), and there was no evidence of impaired immunity or exposure to individuals with known or suspected P. carinii disease. Culture of the lavage fluid also revealed pathogens typical of lung disease associated with cystic fibrosis. It is suspected that the presence of P. carinii in this patient represented a new acquisition, as has been described in immunocompetent infants and children. Whether P. carinii infection complicated cystic fibrosis-associated lung disease in this patient is unknown. PMID- 10686046 TI - Right lung agenesis with left pulmonary artery sling. AB - We report on a 2-month-old infant girl who had right pulmonary agenesis and an unusual course of the left pulmonary artery. Computed tomography and cardiac catheterization showed that the left pulmonary artery arose from the main pulmonary artery, crossing the midline, and reaching the left lung via an aberrant course between the esophagus and trachea. The coexistence of right pulmonary agenesis and left pulmonary sling is extremely rare. Unlike in other reports, our patient remained symptom-free and in good health, with normal growth and development until age 2 years, when she died from complications during an attack of bronchiolitis caused by respiratory syncytial virus. PMID- 10686047 TI - Re: Rost et al. describing the effects of neck position on endotracheal tube (ETT) location in low birth weight infants. PMID- 10686048 TI - The authors respond. PMID- 10686049 TI - Role of progestational agents in the treatment of undernourished patients with cystic fibrosis. PMID- 10686050 TI - Ibuprofen therapy in cystic fibrosis. PMID- 10686051 TI - Response by author. PMID- 10686052 TI - Editorial PMID- 10686053 TI - Experiences in HCV-NAT screening prior to releasing cellular components by the German Red Cross Blood Transfusion Service of Baden-Wurttemberg. AB - In this report we present the accumulated data on nucleic acid testing (NAT) for hepatitis C virus (HCV) RNA of blood donations by the Blood Transfusion Service of Baden-Wurttemberg in the period between March 1997 and March 1999. An extra barcoded blood sample was collected from each donor. Samples were tested by NAT in mini-pools of maximally 96 samples. First-time and repeat donors were tested separately. RT/HCV-PCR was performed with the COBAS HCV Amplicortrade mark, versions 1.0 and 2.0 from Roche Diagnostic Systems. Many modifications have been introduced to the original protocol since the implementation of NAT screening aiming at an increase in the sensitivity and specificity of the assay. NAT positive pools containing serologically positive samples were detected. Initially, reactive pools were identified that could not be confirmed by secondary pooling and single testing procedures. So far, no serologically negative but NAT positive sample has been found. PMID- 10686054 TI - Standardization: a progress report. AB - The introduction of nucleic acid amplification technology (NAT) assays for the detection of viral contamination of blood and blood products requires the availability of well-characterized reference reagents. Working reagents for hepatitis C virus RNA, hepatitis B virus DNA, HIV-1 RNA and human parvovirus B19 DNA have been established at NIBSC and at many other laboratories (both official medicinal control laboratories and commercial laboratories). However, as these reagents have been characterised independently, it is difficult to compare results from assays using different working reagents. Recently, a WHO International Standard was established for HCV RNA NAT assays. This standard has been calibrated in International Units (IU) and provides a common standard against which all working reagents can be calibrated. Collaborative studies to characterise two further candidate International Standards for HBV DNA and HIV-1 RNA NAT assays have been completed. PMID- 10686055 TI - Validation of HCV-NAT assays and experience with NAT application for blood screening in Germany. AB - Validation of HCV-NAT assays is an important prerequisite for the use of NAT for screening plasma or blood donations. The main NAT features to be validated are specificity, detection limit and robustness. Preliminary experience in Germany obtained with different methodical and logistic approaches shows the feasibility of HCV-NAT as a screening test for blood donations. PMID- 10686056 TI - Assessment of validation studies from a fractionator's point of view. AB - NAT testing has become an integral part in the safety programs of both plasma fractionators and transfusion services. NAT testing for HCV RNA is now mandatory for plasma fractionators in Europe and for transfusion services in Germany and Austria. Before NAT testing of plasma could become mandatory, a defined environment had to be created to allow comparison of different NAT procedures. To create such an environment, international virus standards, as well as guidelines for validation, assessment of robustness, and quality assurance of NAT have been released. This paper is a critical review of currently available standards and national reference preparations, detection limits, and national regulations of NAT in view of the specific nature of NAT. PMID- 10686057 TI - European multi-centre validation study of NucliSens Extractor in combination with HCV Amplicor 2.0 assay for HCV-NAT screening of plasma pools. AB - The NucliSens Extractor in combination with the 2.0 version of the Roche Cobas HCV Amplicor test has been validated by five European blood screening laboratories in a multi-centre study. For testing the performance characteristics of this HCV-NAT method, the European Pharmacopoeia validation guidelines were followed. The CLB VQC reference reagents were used for testing robustness and sensitivity. After a technical improvement in the extraction stations, the NucliSens Extractor appeared to be contamination-free as was proved by testing negative controls alternating with samples containing a high HCV-RNA concentration. The Pelicheck HCV-RNA genotype 1 dilution panel was tested 74 times in the five laboratories and an overall 95% detection limit of 80 genome equivalents (geq)/ml was found. In one laboratory the Pelicheck panel was tested in 25 runs and here a 95% detection limit of 32 geq/ml was achieved. In this laboratory the Pelispy HCV-RNA run control samples of 140 geq/ml were consistently picked up in all extractor stations. In addition the laboratories have tested a WHO HCV-RNA genotype 1 standard dilution series 39 times and a Pelicheck HCV-RNA genotype 3 reference panel in 32 test runs. The limiting dilution analysis enabled us to compare the detection efficiency of the NucliSens Amplicor method for the genoype 1 and genotype 3 isolates and to calibrate the reference reagents against each other. The combined Nuclisens-Amplicor method was found to detect the genotype 3 isolate in the Pelicheck HCV-RNA panels with 2-3 fold lower efficiency than the genotype 1 standard (assuming that the historical calibration of the genotype 3 against the genotype 1 standard is correct). In this study of a single method 1 IU of the WHO HCV-RNA standard was found to be equivalent to 5.1 geq of the VQC HCV-RNA standard (95% confidence intervals 3.1 9.1 geq). To avoid confusion with the use of the CLB VQC reagents we accept the NIBSC collaborative study in which calibration by a variety of methods showed that the Pelispy 380 geq/ml run control is equivalent to 100 IU/ml of the WHO standard. This multi-centre validation study demonstrates that the 95% detection limit of the NucliSens HCV Amplicor method lies far below the detection limits required by the international regulatory bodies. PMID- 10686058 TI - Performance characteristics of the AmpliScreen HIV-1 test, an assay designed for screening plasma mini-pools. AB - This study evaluated the performance characteristics of the AmpliScreen(TM)Human Immunodeficiency Virus-Type 1 (HIV-1) Test, Version 1.5, a test designed for screening pools composed of samples from individual units of blood or plasma. HIV 1, hepatitis C (HCV) and hepatitis B (HBV) virus particles were simultaneously extracted and concentrated from plasma by a multi-prep sample processing procedure. An HIV-1 Internal Control (IC) RNA was added to each sample to serve as an extraction and amplification control. Processed samples were amplified by RT-PCR using HIV-1-specific complementary primers and detected by hybridization of the amplified products to HIV-1- and IC-specific oligonucleotide probes. The analytical sensitivity of the test (concentration that yields >/=95% positive results in a set of replicate tests) was 25 copies of HIV-1 RNA per mL of pooled plasma. Representative strains from all HIV-1 group M subtypes were reproducibly detected (>95% positive results among 22 replicate tests) at concentrations of 30 to 75 viral particles per ml. The test exhibited excellent specificity; it did not cross-react with a set of 30 viral and five bacterial isolates and yielded negative results on a panel of 500 blood samples from HIV-1 seronegative donors. Samples containing abnormally high levels of haemoglobin, albumin, triglycerides or bilirubin in plasma samples did not interfere with the detection of HIV-1 RNA at a concentration of 100 copies of per ml. The test detected HIV-1 RNA 7-17 days prior to anti-HIV-1 antibody seroconversion for all 10 seroconversion panels tested. A fully automated COBAS AmpliScreen(TM)version of this test is being validated. COBAS AmpliScreen tests for HCV and HBV also incorporate the multi prep specimen processing method, thereby making it possible to use a single processed specimen to screen for all three viruses. PMID- 10686059 TI - Safety issues for plasma derivatives and benefit from NAT testing. AB - Manufacturing processes for plasma derivatives are in general highly effective for removal or inactivation of enveloped viruses and the products are safe with regard to the clinically important viruses HIV, HCV and HBV. They are not so effective for the elimination for non-enveloped viruses, especially Parvovirus B19 (B19). A certain risk remains of B19 contamination for some plasma derivatives that is caused, firstly, by the occurrence of highly contaminated donations (up to 10(14)genomes/ml) and secondly, by the extreme heat resistance and small size of B19 which makes it difficult to remove or inactivate. NAT is a beneficial tool for detection of virus contamination. It is routinely used for the detection of HCV-RNA in plasma pools, thereby preventing the processing of HCV-RNA positive material. NAT assays may also be valuable for testing the removal of viruses during manufacturing. This may be especially important if a virus cannot be tested by infectivity assays. PMID- 10686060 TI - NAT: perspectives for cellular components. AB - The introduction of routine testing to detect viral genomes in donated blood was originally driven by requirements for plasma fractionation in relation to exclusion of hepatitis C virus (HCV) RNA. Nevertheless, it was obvious from the outset that a dual standard for fractionated products and individual blood components would be untenable. In many countries therefore, planning for introduction of nucleic acid testing (NAT) of blood incorporated progression to release of HCV RNA tested components. HCV was singled out because of its long seronegative 'window period', relatively high prevalence and incidence in blood donors, rapid burst time and high genome copy number during seroconversion. The latter properties made HCV particularly suitable for detection in pools of samples. If HCV RNA testing is required for release of labile components such as platelets, rapid provision of NAT results is vital because of short shelf life of platelets and the problems of delays when resolving the infectious unit in a reactive pool. For NAT release of labile components smaller sample pool sizes allow faster resolution of RNA positive units. Smaller pools involve high test throughput, the likely need for more testing laboratories and ensuing increased costs. Single sample testing is the ultimate extrapolation of reducing sample pool size. With reduced pool sizes or single sample testing, the option of testing for other viruses (e.g. HIV or HBV) singly or in multiplex also arises. The cost-benefit and incremental yield of such strategies in the light of 'combo' assays for HIV Ag/Ab and the recently described HCV Ag assay will require careful and objective assessment, together with re-appraisal of anti-HBc screening for detection of HBV infected donors at the "tail-end" of carriage. PMID- 10686061 TI - Assessment of needs for plasma for fractionation in Europe. AB - In the early 1990s, a series of outbreaks of hepatitis C (HCV) infections clustering among recipients of certain lots of plasma-derived medicinal products (PDMP) alarmed regulatory authorities, manufacturers and the public alike. Also, a few episodes of Hepatitis A (HAV) infections occurred in haemophiliacs receiving solvent-detergent-treated factor VIII concentrates. Thus, several measures were brought into effect to reestablish the safety of the incriminated products and to further increase the margin of safety of PDMP in general. Therefore, intramuscular immunoglobulins had to be free of HCV RNA as shown by nucleic acid amplification technology (NAT) in the final products. Furthermore, the manufacturing process of PDMP had to be validated for both viral inactivation and elimination. Finally, HCV-NAT was to be standardised and implemented as a validated test of plasma pool samples. In 1994, a joint meeting of EPFA, EAPPI and Regulatory Authorities was held in Brussels to outline the state of the art and to delineate the actions to be taken. Five years later, in 1999, the incidence rates of HIV, HBV and HCV in unpaid blood donors have been minimized, especially in European countries. With probabilities for window period donations as low as 0.6 in 1 million for both HIV and HCV and 2.1 in 1 million for HBV in Switzerland, labile blood products have reached extreme, but not absolute safety. The introduction of HCV-NAT roughly doubles this safety resulting in a 1 in 3 million probability of a window donation.Concomittantly, extensive viral validation studies document effective inactivation and removal of viruses in PDMP. The demonstrated margins of safety, expressed as logarithmical reduction factors (LRF), range from 4 to over 20 log(10), depending on product, virus, and inactivation procedure used. Further progress to even safer PDMP shall be acomplished by consolidating the GMP processes, abandoning of obsolete requirements and harmonising national regulations within Europe. Before introducing new measures for additional agents such as HAV or Parvovirus B 19, gains and risks and even potential new threats have to be carefully assessed. Alternative efforts for the safeguard of patients, e.g. vaccination for HAV, need to be balanced against the risks of changing established and validated manufacturing procedures of PDMP with long-lasting safety records. PMID- 10686062 TI - Risk-benefit analysis of nucleic acid testing: the French experience. PMID- 10686063 TI - Sixth EPFA/NIBSC workshop on NAT in rome 1999: outcome of discussions. AB - The sixth international workshop on "Nucleic Acid Amplification Technology for the Detection of Blood Borne Viruses" was held on 5-6 May 1999 at the Istituto Superiore di Sanita in Rome, Italy. The purpose of the workshop was to bring together regulators, blood and plasma product manufacturers, national control agencies, test kits manufacturers and scientists to discuss recent experience as well as regulatory topics related to the use of NAT for the detection of blood borne viruses. Various papers were presented and discussed in 6 sessions. PMID- 10686064 TI - Contents and index to vol. 27 PMID- 10686065 TI - Induction by synaptic zinc of heat shock protein-70 in hippocampus after kainate seizures. AB - Following seizures, heat shock protein (HSP)-70 is induced in various brain regions. Since zinc that can induce HSP-70 in various cell systems is enriched in certain glutamatergic terminals and translocates to postsynaptic neurons with seizures, we examined the possibility that HSP-70 induction in the epileptic brain is mediated by synaptic zinc. Adult rats were injected intraperitoneally with kainate to induce seizures. Seizures were halted 3 h after the kainate administration by the injection of phenytoin. Staining of brain sections with zinc-specific fluorescent dye TFL at 24 h after the kainate injection revealed a one-to-one correlation between dense TFL fluorescence and acidophilic neuronal degeneration in the hippocampus. Subsequent staining with anti-HSP-70 antibody, however, revealed that more numerous neurons than degenerating neurons exhibited HSP-70 immunoreactivity. Most of the HSP-70(+) neurons were not stained with acid fuchsin but exhibited mild zinc fluorescence in the cytoplasm. Intraventricular injection of CaEDTA attenuated neuronal death as well as the HSP-70 induction in a dose-dependent manner. Supporting the specificity of zinc rather than calcium as the inducer of HSP-70 in neurons, exposure to zinc but not to a calcium ionophore or excitotoxins increased expression of HSP-70 mRNA and protein in cultured cortical neurons. The present results suggest that not only selective neuronal death, but also HSP-70 induction in neurons after seizures, is mediated by the translocation of endogenous synaptic zinc. PMID- 10686066 TI - Differential effects of BDNF, ADNF9, and TNFalpha on levels of NMDA receptor subunits, calcium homeostasis, and neuronal vulnerability to excitotoxicity. AB - Calcium influx through N-methyl-d-aspartate (NMDA) receptors can result in neuronal apoptosis or necrosis and may play a pivotal role in neuronal death in many different neurodegenerative diseases. In the present study we employed primary neuronal cultures and three different excitoprotective factors, brain derived neurotrophic factor (BDNF), activity-dependent neurotrophic factor (ADNF9), and tumor necrosis factor alpha (TNFalpha), to elucidate the mechanisms whereby trophic factors modify the excitotoxic process. Neurons pretreated with BDNF exhibited increased levels of the NMDA receptor subunits NR1 and NR2A, which was associated with increased calcium responses to NMDA and vulnerability to excitotoxic necrosis and reduced vulnerability to apoptosis. ADNF9 and TNFalpha suppressed calcium responses to glutamate and protected neurons against both excitotoxic necrosis and apoptosis, but had no effect on levels of NMDA receptor subunits. Inhibition of phosphorylation and DNA binding of NF-kappaB, by H7 and kappaB decoy DNA, respectively, suggest that the excitotoxic-modulating actions of BDNF are mediated by kinases, while those of ADNF9 and TNFalpha are mediated by both kinases and the transcription factor NF-kappaB. Our data show that, whereas BDNF increases neuronal responses to glutamate while ADNF9 and TNFalpha decrease the same, all three protect against excitotoxic apoptosis. PMID- 10686067 TI - Transduction of human GAD67 cDNA into immortalized striatal cell lines using an Epstein-Barr virus-based plasmid vector increases GABA content. AB - The M213-20 and M213-1L cell lines were immortalized from rat striatum using the tsA58 allele of the SV40 large T antigen, contain the GAD enzyme, and produce GABA (Giordano et al., 1994, Exp. Neurol. 124:395-400). Cell lines that produce large amounts of GABA may be useful for transplantation into the brain in conditions such as Huntington's disease or epilepsy, where localized application of GABA may be of therapeutic value. We have explored the potential use of the pREP10 plasmid vector, which replicates episomally, to increase GAD expression and GABA production in M213-20 and M213-1L cells. Human GAD(67) cDNA was transfected into M213-20 and M213-1L, and subclones were isolated with hygromycin selection. Immunochemical studies showed increased GAD(67) expression compared to the parent M213-20 and M213-1L cell lines. Staining for the EBNA antigen and Southern blots demonstrated that the pREP10 plasmid was stably maintained in the cells for at least 12-15 months in culture. Several clones were isolated in which GABA concentrations were increased by as much as 4-fold (M213-1L) or 44-fold (M213-20) compared to the parent cell lines or 12-fold (M213-1L) and 94-fold (M213-20) greater than rat striatal tissue (1.678 +/- 0.4 micromol/g prot). The ability of these cells to continue to produce large amounts of GABA while being maintained in culture for extended periods suggests that similar methods might be used with human cell lines to produce cells that can be transplanted into the brain to deliver GABA for therapeutic purposes. PMID- 10686068 TI - Persistent CREB phosphorylation with protection of hippocampal CA1 pyramidal neurons following temporary occlusion of the middle cerebral artery in the rat. AB - Phosphorylation of the DNA-binding transcription factor, cyclic AMP response element binding protein (CREB), was immunohistochemically examined in rat brain hippocampal CA1 in order to examine the ischemic vulnerability of this region from the viewpoint of CREB activation. The rat brain had been subjected to 90-min focal ischemia followed by various periods of recirculation. Focal ischemia was induced by occlusion of the middle cerebral artery using the intraluminal suture method. CA1 pyramidal neurons in the sham animals showed definite immunoreactivity with anti-CREB antibody, which binds to both unphosphorylated and phosphorylated CREB, while reactivity with anti-phosphorylated CREB antibody was barely detectable in these neurons. In contrast, at 3.5 h of recirculation, a significant increase in the number of phosphorylated CREB-positive neurons was noted in the CA1 on both sides, and the increase continued until 48 h of recirculation with a tendency for gradual decline. At each period, the ischemic side showed a more marked increase in the number of immunoreactive cells as compared to the nonischemic side. Cresyl violet staining revealed CA1 pyramidal neurons to be maintained intact until 14 day of recirculation, at which time CREB phosphorylation has returned to the control level. Transient global ischemia is known to induce only mild CREB phosphorylation in the CA1 followed by a frank neuronal loss in this region. These data suggest that CREB phosphorylation can be persistently activated in CA1 neurons after focal ischemia and that this phenomenon may be closely associated with protection of these neurons. PMID- 10686069 TI - Neuroprotective and neurodestructive functions of nitric oxide after spinal cord hemisection. AB - Nitric oxide (NO) may subserve different functions in different central neurons subjected to axotomy. The difference may depend on whether the neurons basally express neuronal nitric oxide synthase (nNOS), a biosynthetic enzyme of NO. This is supported by our previous finding that suggests the differential role of NO in neurons of nucleus dorsalis (ND) and red nucleus (RN) which have different basal expression of nNOS. This study aimed to establish firmly the functions of NO, as revealed by nNOS immunoreactivity and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry, by the administration of endogenous NO donor, l-arginine (l-arg), and NOS inhibitor, l-N(G)-nitroarginine methyl ester (l-NAME). To relate the role of NO to glutamate receptors (GluR), the distributions of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and N-methyl-d-aspartate receptor (NMDAR) in the two nuclei were revealed by immunohistochemical techniques. nNOS immunoreactivity was void in ND neurons, but expressed weakly in the RN normally. It was induced in ipsilateral ND neurons and upregulated on both sides of RN after spinal cord hemisection. Neuronal loss in the ipsilateral ND was augmented by l-arg, but reduced by l NAME. In the contralateral RN, l-arg attenuated neuronal loss. NMDAR1 was present in most neurons in ND. After axotomy, some NMDAR1 immunoreactive neurons of the ipsilateral ND were induced to express NOS, whereas RN neurons showed strong staining for NMDAR1 and all the AMPA subunits. Most of the NOS-positive neurons in the RN were coexistent with GluR2 in normal rats and those subjected to axotomy. The present data demonstrated that NO exerted neurodestructive function in the non-NOS-containing ND neurons characterized by NMDAR as the predominant glutamate receptor. NO might be beneficial to the NOS-containing RN neurons. This could be attributed to the presence of GluR2. Possible diverse synthesizing pathways of NO in two different central nuclei were suggested from the observation that NOS was colocalized with NADPH-d in ND neurons, but not in RN neurons. PMID- 10686070 TI - Conditionally immortalized cell lines, engineered to produce and release GABA, modulate the development of behavioral seizures. AB - Transplantation of genetically engineered cells can provide sustained focal delivery of naturally occurring molecules, including neurotransmitters and growth factors. We have engineered immortalized mouse cortical neurons and glia to deliver GABA by driving GAD(65) expression. Engineered cell lines showed GAD(65) mRNA expression, enzymatic activity, and GABA release. In vitro, basal flux of GABA was approximately 20% of total cellular GABA. We transplanted these GABA producing cells bilaterally into either the anterior or the posterior substantia nigra of 43 rats. The rats were subsequently kindled through an electrode placed in the entorhinal cortex. GABA-producing cells, but not beta-galactosidase producing cells, affected kindling rates. The number of stimulations needed to reach the first stage-5 seizure and to achieve full kindling differed significantly between the anterior and posterior transplantation sites when GAD(65)-producing cells were transplanted but not when beta-galactosidase producing cells were transplanted. Our data show that transplanted engineered cells can make and release GABA at physiologically meaningful concentrations. PMID- 10686071 TI - An in vivo quantifiable model of cochlear neuronal degeneration induced by central process injury. AB - In the available in vivo experimental models for cochlear neuronal degeneration, the peripheral (hair cell side) process of the cochlear nerve has been injured in order to induce neuronal degeneration. However, there has been no dependable experimental model in which cochlear neuronal degeneration begins from the central (brain stem side) process. This lack of a central process injury model has probably been due to the experimental difficulties that had to be overcome in order to reproducibly and selectively injure the central process of the cochlear neurons while maintaining the patency of the internal auditory artery in small experimental animals such as rats. Using rats, we first developed a central process injury model in which the reduction of the spiral ganglion cells due to retrograde degeneration of cochlear neurons can be quantitatively evaluated. In our experimental model, the cochlear nerve was compressed and injured by a compression-recording (CR) electrode placed at the internal auditory meatus. First, the cochlear nerve was compressed until the compound action potentials of the cochlear nerve became flat, and then the CR electrode was advanced by various compression speeds (5, 10, or 200 micrometer/s) to reach the same depth (400 micrometer). In our model, therefore, the reduction of the spiral ganglion cells was caused compression speed dependently. This method made it possible to produce compression injury to the cochlear nerve without evidence of damage to the blood supply to the cochlea via the internal auditory artery. This model gives us the means to obtain knowledge that was previously impossible to derive from the peripheral process injury models. PMID- 10686072 TI - Sequential administration of GDNF into the substantia nigra and striatum promotes dopamine neuron survival and axonal sprouting but not striatal reinnervation or functional recovery in the partial 6-OHDA lesion model. AB - Glial cell line-derived neurotrophic factor (GDNF) has prominent survival promoting effects on lesioned nigrostriatal dopamine neurons, but understanding of the conditions under which functional recovery can be obtained remains to be acquired. We report here the time course of nigrostriatal axon degeneration in the partial lesion model of Parkinson's disease and the morphological and functional effects of sequential administration of GDNF in the substantia nigra (SN) and striatum during the first 5 weeks postlesion. By 1 day postlesion, the nigrostriatal axons had retracted back to the level of the caudal globus pallidus. Over the next 6 days axonal retraction progressed down to the SN, and during the following 7 weeks 74% of tyrosine hydroxylase-positive (TH(+)) and 84% of retrogradely labeled nigral neurons were lost, with a more pronounced loss in the rostral part of the SN. GDNF administration protected 70 and 72% of the nigral TH(+) and retrogradely labeled cell bodies, respectively, but did not prevent the die-back of the lesioned nigrostriatal axons. Although clear signs of sprouting were observed close to the injection site in the striatum as well as in the globus pallidus, the overall DA innervation of the striatum [as measured by [(3)H]-N-[1-(2-benzo(b)thiopenyl)cyclohexyl]piperidine-binding autoradiography] was not improved by the GDNF treatment. Moreover, the lesion-induced deficits in forelimb akinesia and drug-induced rotation were not attenuated. We conclude that functional recovery in the partial lesion model depends not only on preservation of the nigral cell bodies, but more critically on the ability of GDNF to promote significant reinnervation of the denervated striatum. PMID- 10686073 TI - The neurotoxicity of the venom phospholipases A(2), notexin and taipoxin. AB - The presynaptically active, toxic phospholipases known as notexin and taipoxin are principal components of the venom of the Australian tiger snake and the Australian taipan respectively. The inoculation of the toxins into one hind limb of rats caused, within 1 h, the depletion of transmitter from the motor nerve terminals of the soleus muscle. This was followed by the degeneration of the motor nerve terminals and of the axonal cytoskeleton. By 24 h 70% of muscle fibers were completely denervated. Regeneration and functional reinnervation were almost fully restored by 5 days, but collateral innervation was common in the regenerated muscles, and this abnormality persisted for at least 9 months. The data provide an explanation for both the severity of neuromuscular paralysis that can accompany envenoming bites by tiger snakes and taipans and the difficulty experienced by physicians in managing the envenomed subjects. PMID- 10686074 TI - Abeta42-carboxy-terminal-like immunoreactivity is associated with intracellular neurofibrillary tangles and pick bodies. AB - Co-occurrence of neurofibrillary tangles (NFTs) and beta-amyloid (Abeta) containing plaques is the pathological hallmark of Alzheimer's disease (AD). Much research has been carried out to elucidate the possible interactions of these two characteristic lesions. Here we show by immunohistochemistry that an Abeta42 carboxy-terminal-like epitope (tAbeta42) is associated with NFTs at an early, intraneuronal stage. The NFTs immunoreactive for tAbeta42 were also positive for tau2. tAbeta42 NFTs occurred even in cases without plaques or any other form of extracellular Abeta deposits. NFTs were tAbeta42 immunoreactive under all tangle forming conditions studied: AD, the parkinsonism dementia complex of Guam, progressive supranuclear palsy, and elderly controls. In Pick disease, which is another "tauopathy" Pick bodies and ballooned neurons were found to be immunoreactive for tAbeta42. Appearance of an epitope of Abeta in neurons at such an early stage of tangle formation and in Pick bodies may indicate a direct connection of Abeta42 and cytoskeletal disarrangement, although cross-reactivity with other epitopes cannot be ruled out. PMID- 10686075 TI - Survival of grafted fetal neural cells in kainic acid lesioned CA3 region of adult hippocampus depends upon cell specificity. AB - We hypothesize that the degree of graft cell survival within the damaged CNS correlates with the specificity of donor cells to the region of grafting. We investigated graft cell survival following transplantation of fetal micrografts into the CA3 region of the adult rat hippocampus at a time-point of 4 days after an intracerebroventricular administration of kainic acid (KA). Grafts consisted of 5'-bromodeoxyuridine (BrdU) labeled embryonic day (E) 19 cells from hippocampal fields CA3 and CA1 and E15 and E19 cells from the striatum. Absolute cell survival in these grafts was quantitatively analyzed at 1 month postgrafting, using BrdU immunostaining of serial sections and three-dimensional reconstruction of grafts. Absolute graft cell survival in lesioned CA3 was dramatically greater for cells having hippocampal origin (CA3 cells, 69% cell survival; CA1 cells, 42% cell survival) than those having nonhippocampal origin, such as striatal cells (E15 cells, 12% cell survival; E19 cells, 4% cell survival). This difference is in sharp contrast to survival of these cells in culture, where E19 cells from both hippocampal and nonhippocampal origins exhibited similar survival. Comparison of survival among hippocampal cell types indicated significantly greater survival for cells that are specific to the lesioned area (i.e., CA3 cells) than for those that are nonspecific to the lesioned area (i.e., CA1 cells). Graft cell survival in the intact CA3 region (contralateral to KA administration), however, did not differ either between cells having hippocampal and nonhippocampal origins or between CA3 and CA1 cells (CA3 cells, 26% cell survival; CA1 cells, 33% cell survival; and E15 striatal cells, 20% cell survival). These results underscore the finding that enhanced survival of fetal cell grafts in the lesioned CNS is critically dependent upon the specificity of donor fetal cells to the region of transplantation. Thus, grafting of cells that are specific to the lesioned area is a prerequisite for achieving maximal graft cell survival and integration in the lesioned host CNS. PMID- 10686076 TI - Netrin-1 and peripheral nerve regeneration in the adult rat. AB - Axonal guidance during development of the nervous system is thought to be highly regulated through interactions of axons with attractive, repulsive, and trophic cues. Similar mechanisms regulate axonal regeneration after injury. The netrins have been shown to influence the guidance of several classes of developing axons. Although netrins have been implicated as axonal guidance cues in the developing peripheral nervous system, there has been no direct evidence of netrin-1 expression in either developing or adult peripheral nerve. The present study utilized competitive PCR and immunohistochemistry to demonstrate the localization of netrin-1 within adult rat sciatic nerve. The expression of netrin-1 mRNA and protein was compared for normal or regenerated sciatic nerve 2 weeks following either a crush or a transection and repair injury. The PCR data show that netrin 1 mRNA is normally expressed at low levels in peripheral nerve, and similar low levels are found 2 weeks following a crush injury. However, 2 weeks following nerve transection and repair there is approximately a 40-fold increase in netrin 1 mRNA levels. Immunohistochemistry data show that Schwann cells are the major source of netrin-1 protein in peripheral nerve. Our results suggest that netrin-1 mRNA levels are profoundly affected during peripheral nerve injury and regeneration. The localization of netrin-1 to Schwann cells suggests that this protein is strategically situated to influence axon regeneration in adult peripheral nerve. PMID- 10686077 TI - Nerve guides seeded with autologous schwann cells improve nerve regeneration. AB - This study evaluates the ability of Schwann cells (SCs) transplanted into a nerve guide to improve regeneration and reinnervation after sciatic nerve resection and repair, leaving a 6-mm gap, in the mouse. SCs were isolated from predegenerated adult sciatic nerves and expanded in culture using a chemically defined medium. Syngeneic, isogeneic, and autologous SCs were suspended in Matrigel and seeded in resorbable, permeable poly(l-lactide-co-epsilon-caprolactone) guides at 150,000 cells/tube. Guides containing SCs were compared to guides filled with Matrigel alone and with peroneal nerve autografts. Functional reinnervation was assessed by noninvasive methods to determine recovery of sweating, nociceptive, sensory, and motor functions in the hindpaw during 4 months postoperation. Morphological analysis of the regenerated nerves was performed at the end of follow-up. The group with an autograft achieved faster and higher levels of reinnervation and higher number of regenerated myelinated fibers than groups repaired by tubulization. The immunogenicity of transplanted SCs influenced the outcome of nerve regeneration. Transplants of autologous SCs resulted in slightly lower levels of reinnervation than autografts, but higher recovery and number of regenerated fibers reaching the distal nerve than transplants of isologous and syngeneic SCs, although most of the differences were not statistically significant. Syngeneic SCs did not improve regeneration with respect to acellular guides. Prelabeled transplanted SCs were found to survive into the guide 1-3 months after implantation, to a larger number when they were autologous than syngeneic. Cellular prostheses composed of a resorbable guide seeded with autologous SCs appear as an alternative for repairing long gaps in injured nerves, approaching the success of autografts. PMID- 10686078 TI - Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody. AB - The presence of the intermediate filament protein nestin has been the predominant marker used to describe stem and progenitor cells in the mammalian CNS. In this study, a 998-bp fragment in the 3' region of the nestin mRNA was cloned from human fetal brain cells (HFBC). The nucleotide sequence of the cloned cDNA revealed 21 differences with the previously published human nestin sequence, resulting in 17 amino acid changes. A 150-amino-acid fragment derived from the cloned nestin cDNA was coupled to glutathione S-transferase and used as an immunogen to generate a rabbit polyclonal antiserum that selectively detects human nestin. HFBC that proliferated in response to basic fibroblast growth factor incorporated 5-bromo-2'-deoxyuridine into their nuclei and immunostained for nestin, indicating nestin expression in proliferating CNS progenitor cells. In all cell cultures, nestin costained with the neuroepithelial cell marker vimentin. A small subset of nestin-stained cells (1-2%) immunostained with neuronal marker MAP-2 during the first week and after 4 weeks in culture. However, during the first week in culture, approximately 10-30% of the total cell population of HFBC stained for the glial cell marker GFAP, and nearly all coimmunostained for nestin. After 4 weeks in culture, a subset of GFAP-positive cells emerged that no longer costained with nestin. These results describe nestin expression not only in CNS progenitor cells but also in the cells which were in transition from a progenitor stage to glial differentiation. Collectively, these data suggest a differential temporal regulation of nestin expression during glial and neuronal cell differentiation. PMID- 10686079 TI - Contraction-induced muscle fiber damage is increased in soleus muscle of streptozotocin-diabetic rats and is associated with elevated expression of brain derived neurotrophic factor mRNA in muscle fibers and activated satellite cells. AB - The expression of brain-derived neurotrophic factor (BDNF) is elevated in the soleus muscle of streptozotocin-diabetic rats. To determine whether this diabetes induced elevation was associated with or enhanced by muscle activity we have induced high-intensity muscle contraction by electrically stimulating the sciatic nerve. In 6-week diabetic rats, intense contraction of the soleus muscle resulted in a two- to four-fold elevation of BDNF mRNA and increased plasma levels of creatine kinase that were associated with severe focal muscle fiber damage and concomitant satellite cell activation. Focal muscle fiber damage and concomitant satellite cell activation were also observed in the soleus muscle of nonstimulated diabetic rats, but to a much lesser extent. No effects of muscle contraction, i.e., experimentally induced or during normal daily activity, on muscle fiber structure or BDNF mRNA expression were seen in diabetic extensor digitorum longus (EDL) muscle. Using a nonradioactive in situ hybridization technique for electron microscopy, the elevated expression of BDNF mRNA in the diabetic soleus muscle was localized within muscle fibers as well as activated satellite cells. This study shows that diabetic soleus muscle, in contrast to diabetic EDL and to soleus and EDL muscle of normal animals, is highly susceptible to contraction-induced damage. Intense contraction and the associated muscle fiber damage in the diabetic soleus muscle result in an upregulation of BDNF mRNA in muscle fibers and activated satellite cells, which may be involved in the restoration and/or maintenance of nerve/muscle integrity. PMID- 10686080 TI - Nicotine attenuates arachidonic acid-induced overexpression of nitric oxide synthase in cultured spinal cord neurons. AB - Primary spinal cord trauma can initiate a cascade of pathophysiologic events which markedly contribute to the expansion and amplification of the primary insult. The detailed mechanisms of these secondary neurochemical reactions are largely unknown; however, they involve membrane lipid derangements with the release of free fatty acids, in particular, arachidonic acid (AA). AA can induce several injury effects on spinal cord neurons. We hypothesize that upregulation of nitric oxide synthase (NOS) is among the most important mechanisms of arachidonic-acid-induced neuronal dysfunction and that nicotine can attenuate this effect. To study these hypotheses, spinal cord neurons were exposed to AA and/or nicotine, and several markers of neuronal nitric oxide synthase (nNOS) metabolism were measured. In addition, cotreatments with either inhibitors of nicotinic receptors or inhibitors of specific NOS isoforms were employed. Treatment with AA markedly increased activity of nNOS, as well as mRNA and protein levels of this enzyme. Changes in nNOS expression were accompanied by an increase in cellular cGMP and medium nitrite levels. Pretreatment with nicotine decreased AA-induced overexpression of nNOS and elevation of nitrite levels. In addition, it appeared that these nicotine effects could be partially modulated both by the alpha7 nicotinic receptors or by nonreceptor mechanisms. Alternatively, the observed changes could also be mediated by an alternate nicotinic receptor mechanism which is not blocked by alpha-bungarotoxin or mecamylamine. Results of the present study indicate that exposure to AA can lead to induction of nNOS in cultured spinal cord neurons. In addition, nicotine can exert a neuroprotective effect by attenuation of AA-induced upregulation of nNOS metabolism. These data may have therapeutic implications for the treatment of acute spinal cord trauma. PMID- 10686081 TI - Oxidized high-density lipoprotein induces neuron death. AB - High-density lipoprotein (HDL) exists within the brain and is highly vulnerable to oxidative modifications. The focus of the present study was to determine the effect of HDL and oxidized HDL (oxHDL) upon neurons, astrocytes, and microglia. Administration of highly oxidized HDL, but not native, minimally, or moderately modified HDL resulted in a dose- and time-dependent increase in oxidative stress and death of cultured rat embryonic neurons. Astrocyte and microglia cultures treated with highly oxidized HDL displayed increased reactive oxygen species formation but no toxicity. Application of oxHDL exacerbated oxidative stress and neuron death induced by beta-amyloid peptide. Studies using pharmacological inhibitors implicate the involvement of calcium and reactive oxygen species in oxHDL-induced neuronal loss. Neural cells expressing increased levels of BCL-2 had decreased levels of oxidative stress and neuron death following exposure to oxHDL. Together, these data demonstrate that oxHDL increases oxidative stress in neurons, astrocytes, and microglia which ultimately culminate in neuron death. PMID- 10686082 TI - Continuous infusion of cyclosporin A postinjury significantly ameliorates cortical damage following traumatic brain injury. AB - Traumatic brain injury (TBI) results in the rapid necrosis of cortical tissue at the site of injury. In the ensuing hours and days, secondary injury exacerbates the original damage resulting in significant neurological dysfunction. Recent reports from our lab demonstrate that a bolus injection of the immunosuppressant cyclosporin A (CsA) is neuroprotective following TBI. CsA transiently inhibits the opening of the mitochondrial permeability transition pore and maintains calcium homeostasis in isolated mitochondria. The present study utilized a unilateral controlled cortical impact model of TBI to assess whether the neuroprotective effects of CsA could be extended by chronic infusion. Adult rats were subjected to a moderate (2 mm) cortical deformation and the extent of cortical damage was assessed using modern stereological techniques. Animals were administrated a 20 mg/kg intraperitoneal bolus of CsA or vehicle 15 min postinjury and osmotic minipumps were implanted subcutaneously to deliver CsA (4.5 or 10 mg/kg/day) or vehicle. All animals receiving CsA demonstrated a significant reduction in lesion volume, with the highest dose offering the most neuroprotection (74% reduction in lesion volume). These results extend our previous findings and demonstrate that chronic infusion of CsA is neuroprotective following TBI. These findings also suggest that the mechanisms responsible for tissue necrosis following TBI are amenable to manipulation. PMID- 10686083 TI - NMDA receptors modulate dopamine loss due to energy impairment in the substantia nigra but not striatum. AB - Defects in energy metabolism have been detected in patients with Parkinson's disease and have been proposed as a contributing factor in the disease. Previous in vitro studies showed that NMDA receptors contribute to the loss of dopamine neurons caused by the metabolic inhibitor malonate. In vivo, it is not known whether this interaction occurs through a postsynaptic action on the cell body in the substantia nigra or through a presynaptic action at the dopamine terminal in the striatum. So we could discern the anatomical level of NMDA receptor involvement, rats were infused with malonate, either into the left striatum or into the left substantia nigra. NMDA receptors were locally blocked by an intranigral or intrastriatal coinfusion of malonate plus MK-801 followed by a second infusion of MK-801 3 h later. Animals were examined at 1 week for striatal and nigral dopamine and GABA levels. Intranigral infusion of malonate (0.5 micromol) produced an approximate 50% loss of both nigral dopamine and GABA. MK 801 (0.1 micromol) provided significant protection against both nigral dopamine and GABA loss and against anterograde damage to dopamine terminals in the striatum. Intrastriatal administration of malonate (2 micromol) produced a 68 and 35% loss of striatal dopamine and GABA, respectively. In contrast to intranigral administration, intrastriatal blockade of NMDA receptors did not protect against striatal dopamine loss, although GABA loss was significantly attenuated. Core body temperature monitored several hours throughout the experiment was unchanged. Consistent with a lack of effect of NMDA antagonists on malonate-induced toxicity to dopamine neurons in striatum, intrastriatal infusion of NMDA had a pronounced effect on long-term GABA toxicity with little effect of dopamine loss. These findings are consistent with a postsynaptic action of NMDA receptors on mediating toxicity to dopamine neurons during impaired energy metabolism. PMID- 10686084 TI - Loss of cholinergic phenotype in basal forebrain coincides with cognitive decline in a mouse model of Down's syndrome. AB - Mice with segmental trisomy of chromosome 16 (Ts65Dn) have been used as a model for Down's syndrome. These mice are born with a normal density of basal forebrain cholinergic neurons but, like patients with Down's syndrome, undergo a significant deterioration of these neurons later in life. The time course for this degeneration of cholinergic neurons has not been studied, nor is it known if it correlates with the progressive memory and learning deficits described. Ts65Dn mice that were 4, 6, 8, and 10 months old were sacrificed for evaluation of basal forebrain morphology. Separate groups of mice were tested on visual or spatial discrimination learning and reversal. We found no alterations in cholinergic markers in 4-month-old Ts65Dn mice, but thereafter a progressive decline in density of cholinergic neurons, as well as significant shrinkage of cell body size, was seen. A parallel loss of staining for the high-affinity nerve growth factor receptor, trkA, was observed at all time points, suggesting a biological mechanism for the cell loss involving this growth factor. Other than transient difficulty in learning the task requirements, there was no impairment of trisomic mice on visual discrimination learning and reversal, whereas spatial learning and reversal showed significant deficits, particularly in the mice over 6 months of age. Thus, the loss of ChAT-immunoreactive neurons in the basal forebrain was coupled with simultaneous deficits in behavioral flexibility on a spatial task occurring for the first time around 6 months of age. These findings suggest that the loss of cholinergic function and the simultaneous decrease in trkA immunoreactivity in basal forebrain may directly correlate with cognitive impairment in the Ts65Dn mouse PMID- 10686086 TI - Growth factors in combination, but not individually, rescue rd mouse photoreceptors in organ culture. AB - The rd mouse retina is an animal model for human retinal dystrophy in which the rod photoreceptors undergo apoptosis during the first 4 weeks in vivo or in organ culture. We have examined the effect of different families of trophic factors on the survival of rd mouse photoreceptors in organ culture. Retinas were harvested from rd mice at postnatal day 2 and grown in organ culture for 27 days in vitro (DIV) in DMEM with 10% fetal calf serum. Ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), fibroblast growth factor-2 (FGF2), glial cell line-derived neurotrophic factor (GDNF), neurturin, and persephon were added individually or in combination to the medium at a dose of 50 ng/ml or less. CNTF + BDNF in combination resulted in photoreceptor survival comparable to wild type retinas after 27 DIV. CNTF + FGF2 or CNTF + GDNF produced a partial prevention of photoreceptor death. Photoreceptor degeneration was not blocked by any of the trophic factors added individually. A significant increase in photoreceptor survival was seen with forskolin added to CNTF, but not to BDNF, FGF2, or GDNF. These results demonstrate that trophic factors promote photoreceptor survival through a synergistic interaction. Increased understanding of receptor interactions and signaling pathways may lead to a potential therapeutic role for combinatorial trophic factors in treatment of photoreceptor dystrophies. PMID- 10686085 TI - TGFbeta trophic factors differentially modulate motor axon outgrowth and protection from excitotoxicity. AB - Transforming growth factor (TGF) beta-like trophic factors have been shown to be protective in acute neuronal injury paradigms. In the current study, we analyzed and compared members of this growing family, including glial cell line-derived neurotrophic factor (GDNF), neurturin, nodal, persephin, and TGFbeta1, for protection against chronic glutamate toxicity. In parallel, we developed a organotypic spinal cord culture system to study the ability of these factors to promote motor axon outgrowth across white matter. Using these systems, we were able to differentiate the neuroprotective effect of the TGFbeta-like factors from their motor axon outgrowth-promoting activity. GDNF, neurturin, persephin, nodal, and TGFbeta1 all protected against excitotoxic motor neuron degeneration. Low amounts of GDNF (1 ng/ml) and high concentrations of neurturin induced vigorous motor axon outgrowth. In contrast, nodal, persephin, and TGFbeta1 did not induce motor axon outgrowth. Both GDNF and neurturin bind to Ret receptor complexes and were capable of activating the MAP kinase pathway. A specific inhibitor of MAP kinase kinase, PD98059, inhibited the motor axon outgrowth-promoting activity of the GDNF but not the neuroprotective activity. Similarly, the specific PI3K inhibitors, LY294002 and wortmannin, were able to inhibit the promotion of motor axon outgrowth by GDNF, but did not affect neuroprotective activity. Our results suggest that the neurite outgrowth-promoting effect of GDNF is mediated through the PI3K and MAP kinase pathways. The neuroprotective effect of GDNF appears to be through a separate pathway. PMID- 10686087 TI - Differential vulnerability of hippocampus, basal ganglia, and prefrontal cortex to long-term NMDA excitotoxicity. AB - In human brain, nonartherosclerotic calcification is associated with normal aging and several pathological conditions without any clear significance. In all situations, calcification appears predominantly in the basal ganglia, but is also frequent in the hippocampus and cerebral cortex. alpha-Amino-(3-hydroxi-5-methyl 4-isoxazol-4-il)-propionic acid-induced lesion of the globus pallidus is associated in rats with the formation of calcium deposits similar to those observed in the human brain. To determine whether direct neuronal activation may induce calcification, N-methyl-d-aspartate (NMDA) was microinjected in rat hippocampus, globus pallidus, and lateral prefrontal cortex. Two months later, neuronal death was associated with calcium deposits that were characterized in terms of distribution and size. A unique population of deposits was present in the hippocampus and prefrontal cortex, whereas in the globus pallidus two main groups could be differentiated. Calcification was always associated with a significant microglial reaction as shown by the peripheral benzodiazepine receptor autoradiography. Monoamine oxidase B autoradiography, reflecting the astroglial reaction, was also significantly increased. Our results provide evidence that acute NMDA neuronal activation leads with time to calcification associated with a glial reaction and indicate that nonartherosclerotic calcification in the human brain may develop from an acute NMDA receptor activation. A key role of the metabotropic mGluR1 receptor is also suggested. PMID- 10686088 TI - The MCT ketogenic diet: effects on animal seizure models. AB - Male Wistar rat pups were weaned at 20 days of age and placed on either a control diet or a ketogenic diet containing medium-chain triglyceride (MCT) oil. After 10 days on the diets, they were subjected to one of four seizure tests-maximal electric shock, threshold electroconvulsive shock, threshold pentylenetetrazol, or maximal pentylenetetrazol. After testing, subjects were sacrificed and blood samples were analyzed for beta-hydroxybutyrate concentration. It was found that the MCT diet produced blood levels of beta-hydroxybutyrate that were comparable to or higher than those commonly reported in clinical studies. However, no anticonvulsant effects were seen in any of the seizure tests. In fact, the tests involving maximal seizures actually showed proconvulsant effects. It appears that clinical levels of ketones may be present in the bloodstream without suppressing seizures. PMID- 10686089 TI - Chronic moderate hyperammonemia impairs active and passive avoidance behavior and conditional discrimination learning in rats. AB - The cerebral dysfunction associated with hepatic encephalopathy is generally considered to have hyperammonemia as one of its main causes. Hyperammonemia impairs the neuronal glutamate-nitric oxide-cyclic GMP pathway and the induction of NMDA receptor-dependent long-term potentiation in the hippocampus. We studied the performance of pre/neonatally and postnatally exposed rats to hyperammonemia on active avoidance, passive avoidance, and conditional discrimination tasks. Pre/neonatal hyperammonemia slowed learning of active avoidance behaviors and impaired memory for the passive avoidance task while postnatal hyperammonemia impaired learning on the conditional discrimination task. Hyperammonemia thus may produce cognitive disturbances that relate to the effects of ammonia on the neuronal glutamate-nitric oxide-cyclic GMP pathway. PMID- 10686090 TI - Selective susceptibility to inhibitors of GABA synthesis and antagonists of GABA(A) receptor in rats with genetic absence epilepsy. AB - Thalamocortical spike-and-wave discharges characterize the nonconvulsive absence seizures that occur spontaneously in genetic absence epilepsy rats from Strasbourg (GAERS), a selected strain of Wistar rats. GABA is crucial in the generation of absence seizures. The susceptibility to convulsions induced by threshold doses of various GABA receptor antagonists and inhibitors of GABA synthesis, kainic acid and strychnine, was compared in GAERS and in nonepileptic rats from a selected control strain (NE). The brain structures involved in the drug-elicited convulsive seizures were mapped by c-Fos immunohistochemistry. Injection of various antagonists of the GABA(A) receptor, bicuculline and picrotoxin, and inverse agonists of the benzodiazepine site (FG 7142 and DMCM) induced myoclonic spike-and-wave discharges followed by clonic or tonic-clonic seizures with high paroxysmal activity on the cortical EEG. The incidence of the convulsions was dose-dependent and was higher in GAERS than in NE rats. Mapping of c-Fos expression showed that the frontoparietal cortex was constantly involved in the convulsive seizures elicited by a threshold convulsant dose, whereas limbic participation was variable. In contrast, GAERS were less susceptible than NE rats to the tonic-clonic convulsions induced by the inhibitors of glutamate decarboxylase, isoniazide and 3-mercaptopropionic acid. The GABA(B) receptor antagonist CGP 56999 and kainic acid induced a similar incidence of seizures in GAERS and NE rats and predominantly activated the hippocampus. No difference in the tonic seizures elicited by strychnine could be evidenced between the strains. These results suggest that an abnormal cortical GABAergic activity may underlie absence seizures in GAERS. PMID- 10686091 TI - Effects of pertussis toxin and galpha-protein-specific antibodies on phosphoinositide hydrolysis in rat brain membranes after cholinergic denervation and hippocampal sympathetic ingrowth. AB - Cholinergic denervation of the hippocampal formation, via medial septal lesions, induces peripheral noradrenergic fibers, originating from the superior cervical ganglion, to grow into the hippocampus. We have previously reported that cholinergic denervation and hippocampal sympathetic ingrowth differentially affect guanosine-5'-O-(3-thiotriphosphate)- as well as guanosine-5'-O-(3 thiotriphosphate) + carbachol-stimulated polyphosphoinositide hydrolysis, suggesting an alteration in G proteins and/or the entire receptor complex. To examine the type of G protein which may be involved in these effects, rat dorsal hippocampal membranes were preincubated with pertussis toxin in the presence of guanosine-5'-O-(3-thiotriphosphate) and guanosine-5'-O-(3-thiotriphosphate) + carbachol. Pertussis toxin reduced guanosine-5'-O-(3-thiotriphosphate) in all groups, while guanosine-5'-O-(3-thiotriphosphate) + carbachol-stimulated phosphoinositide hydrolysis was reduced in controls and animals without sympathetic ingrowth but not in animals with hippocampal sympathetic ingrowth. This suggests that pertussis toxin-sensitive G proteins may be involved in the mediation of phosphoinositide hydrolysis. To confirm this hypothesis, membranes were preincubated with antibodies to Galphao and Gq/11. The Go antibody significantly decreased guanosine-5'-O-(3-thiotriphosphate) in all groups, while guanosine-5'-O-(3-thiotriphosphate) +carbachol-stimulated phosphoinositide hydrolysis was reduced only in hippocampal sympathetic ingrowth. Impairment of guanosine-5'-O-(3-thiotriphosphate) and carbachol-stimulated phosphoinositide hydrolysis was also decreased in all groups when preincubated with Gq/11 antibody. To determine whether hippocampal sympathetic ingrowth or cholinergic denervation altered the concentration of various G proteins, immunoblotting methodology was utilized. Gq/11 concentrations were found to be equivalent among groups. The density of Go1, Go2, and Go3 isoforms was significantly increased in the cholinergic denervation, while in the hippocampal sympathetic ingrowth only group Go3 was significantly increased. When assessed as total Go protein, density was increased significantly only in the cholinergic denervation group. Overall, these results suggest that hippocampal sympathetic ingrowth and cholinergic denervation induce alterations in phosphoinositide hydrolysis through both the Gq/11 and the Go proteins and that the coupling between muscarinic receptor and G protein is the possible site which affects changes in phosphoinositide turnover. Our results also suggest that cholinergic denervation and hippocampal sympathetic ingrowth may mediate phosphoinositide hydrolysis through an effect on different isoforms of the same G protein. PMID- 10686092 TI - Evaluating the efficacy of citicoline in embolic ischemic stroke in rats: neuroprotective effects when used alone or in combination with urokinase. AB - The combination of thrombolysis with neuroprotection, because of different mechanisms, would be expected to show better results when used after onset of focal ischemia. In this study we report our experience with the neuronal protective effects of citicoline alone and in combination with urokinase in a model of focal ischemia. Both medications were injected 2 h after onset of a focal occlusion of the middle cerebral artery (MCA) in rats. Focal ischemia was produced with embolization of a clot into the origin of the MCA. This produces a large infarction involving the cortex and the basal ganglia. Animals were observed for neuronal deficts at 2 and 24 h after surgery and were sacrificed 72 h after onset of ischemia. Saline-treated animals showed a large infarction involving the cerebral cortex and basal ganglion in most animals (volume 33.1 +/- 9.7%). Animals treated with citicoline alone were divided in two groups. The first group of animals were treated with a single injection (300 mg/kg, ip) of the medication 2 h after the arterial occlusion. The second group was treated with the active medication intermittently (3 x 300 mg/kg, ip) over a 72-h period. There was a significant decrease in the neuronal damage in the cortex in the animals treated with citicoline (single dose, 20.9 +/- 9.7%, P = 0.01; intermittent injection, 18.9 +/- 11.4%, P < 0.008). The last experiment evaluated the usefulness of the combination of citicoline with intraarterial urokinase. The combination showed significantly more protection than with urokinase or citicoline alone (volume 13.6 +/- 9.1%, P < 0.001). We conclude from our experiments that citicoline may offer significant neuronal protection that may be further enhanced with the addition of a thrombolytic agent. PMID- 10686093 TI - Cold exposure enhances tactile allodynia transiently in mononeuropathic rats. AB - A laser and erythrosin-B-induced sciatic nerve injury decreases thresholds of a mechanically induced paw withdrawal reflex and enhances cold-induced withdrawal behavior of the affected limb. Exposure of the affected paw to a normally innocuous cold stimulus results in a transient decrease in the threshold of the mechanically evoked paw withdrawal reflex in neuropathic but not in intact rats. The present data suggest that in an experimental neuropathic state a normally innocuous cold stimulus may further sensitize spinally mediated withdrawal reflexes to stimuli of another stimulus modality, in this case, to innocuous tactile stimuli. Therefore, testing mechanical allodynia in neuropathic rats immediately after testing cold allodynia may produce artifactual results. PMID- 10686094 TI - Cryo-trapping the six-coordinate, distorted-octahedral active site of manganese superoxide dismutase. AB - Superoxide dismutase protects organisms from potentially damaging oxygen radicals by catalyzing the disproportionation of superoxide to oxygen and hydrogen peroxide. We report the use of cryogenic temperatures to kinetically capture the sixth ligand bound to the active site of manganese superoxide dismutase (MnSOD). Synchrotron X-ray diffraction data was collected from Escherichia coli MnSOD crystals grown at pH 8.5 and cryocooled to 100 K. Structural refinement to 1.55 A resolution and close inspection of the active site revealed electron density for a sixth ligand that was interpreted to be a hydroxide ligand. The six-coordinate, distorted-octahedral geometry assumed during inhibition by hydroxide is compared to the room temperature, five-coordinate, trigonal bipyramidal active site determined with crystals grown from practically identical conditions. The gateway residues Tyr34, His30 and a tightly bound water molecule are implicated in closing-off the active site and blocking the escape route of the sixth ligand. PMID- 10686095 TI - Nature disfavors sequences of alternating polar and non-polar amino acids: implications for amyloidogenesis. AB - Recent experiments with combinatorial libraries of de novo proteins have demonstrated that sequences designed to contain polar and non-polar amino acid residues arranged in an alternating pattern form fibrillar structures resembling beta-amyloid. This finding prompted us to probe the distribution of alternating patterns in the sequences of natural proteins. Analysis of a database of 250,514 protein sequences (79,708,024 residues) for all possible binary patterns of polar and non-polar amino acid residues revealed that alternating patterns occur significantly less often than other patterns with similar compositions. The under representation of alternating binary patterns in natural protein sequences, coupled with the observation that such patterns promote amyloid-like structures in de novo proteins, suggests that sequences of alternating polar and non-polar amino acids are inherently amyloidogenic and consequently have been disfavored by evolutionary selection. PMID- 10686096 TI - Antirestriction protein Ard (Type C) encoded by IncW plasmid pSa has a high similarity to the "protein transport" domain of TraC1 primase of promiscuous plasmid RP4. AB - The IncW plasmid pSa contains the gene ard encoding an antirestriction function that is specific for type I restriction and modification systems. The nucleotide sequence of ard was determined and an appropriate polypeptide of about 33 kDa was identified in Escherichia coli T7 expression system. Analysis of deduced amino acid sequence of Ard encoded by pSa revealed that this protein has no significant similarities with the known Ard proteins (ArdA and ArdB types) except the "antirestriction" motif (14 amino acid residues in length) conserved for all known Ard proteins. This finding suggests that pSa Ard may be classified as a new type of Ard proteins which we designated ArdC. The remarkable feature of ArdC is that it has a high degree of similarity (about 38 % identity) to the N-terminal region of RP4 TraC1 primase which includes about 300 amino acid residues and seems to be essential for binding to the single-stranded DNA and TraC1 protein transport to the recipient cells during the conjugal transfer of plasmid DNA. ArdC also binds to single-stranded DNA. In addition, this protein is able in vitro to protect the single-stranded but not double-stranded plasmid DNA against the activity of type II restriction endonuclease HhaI that cleaves both single and double-stranded DNA. We suggest that like TraC1, ArdC would be transported as a result of their interaction with the single-stranded DNA of transferred plasmid strand during conjugative passage through the cell envelope to the recipient bacterium. Such properties of ArdC protein might be useful to protect immediately the incoming single-stranded DNA from the host endonucleases. PMID- 10686097 TI - Sequence and position-dependence of the equilibrium accessibility of nucleosomal DNA target sites. AB - We have previously shown that nucleosomes are conformationally dynamic: DNA sequences that in the time-average are buried inside nucleosomes are nevertheless transiently accessible, even to large proteins (or any other macromolecule). We refer to this dynamic behavior as "site exposure". Here we show that: (i) the equilibrium constants describing this dynamic site exposure decrease progressively from either end of the nucleosomal DNA in toward the middle; and (ii) these position-dependent equilibrium constants are strongly dependent on the nucleosomal DNA sequence. The progressive decrease in equilibrium constant with distance inside the nucleosome supports the hypothesis that access to sites internal to a nucleosome is provided by progressive (transient) release of DNA from the octamer surface, starting from one end of the nucleosomal DNA. The dependence on genomic DNA sequence implies that a specific genomic DNA sequence could be a major determinant of target site occupancies achieved by regulatory proteins in vivo, by either governing the time-averaged accessibility for a given nucleosome position, or biasing the time-averaged positioning (of mobile nucleosomes), which in turn is a major determinant of site accessibility. PMID- 10686098 TI - Structural and functional comparative study of the complexes formed by viral o29, Nf and GA-1 SSB proteins with DNA. AB - Single-stranded DNA-binding proteins have in common their crucial roles in DNA metabolism, although they exhibit significant differences in their single stranded DNA binding properties. To evaluate the correlation between the structure of different nucleoprotein complexes and their function, we have carried out a comparative study of the complexes that the single-stranded DNA binding proteins of three related bacteriophages, o29, Nf and GA-1, form with single-stranded DNA. Under the experimental conditions used, o29 and Nf single stranded DNA-binding proteins are stable monomers in solution, while GA-1 single stranded DNA-binding protein presents a hexameric state, as determined in glycerol gradients. The thermodynamic parameters derived from quenching measurements of the intrinsic protein fluorescence upon single-stranded DNA binding revealed (i) that GA-1 single-stranded DNA-binding protein occludes a larger binding site (n=51 nt/oligomer) than o29 and Nf SSBs (n=3.4 and 4.7 nt/monomer, respectively); and (ii) that it shows a higher global affinity for single-stranded DNA (GA-1 SSB, K(eff)=18.6 x 10(5) M(-1); o29 SSB, K(eff)=2.2 x 10(5) M(-1); Nf SSB, K(eff)=2.9 x 10(5) M(-1)). Altogether, these parameters justify the differences displayed by the GA-1 single-stranded DNA-binding protein and single-stranded DNA complex under the electron microscope, and the requirement of higher amounts of o29 and Nf single-stranded DNA-binding proteins than of GA-1 SSB in gel mobility shift assays to produce a similar effect. The structural differences of the nucleoprotein complexes formed by the three single stranded DNA-binding proteins with single-stranded DNA correlate with their different functional stimulatory effects in o29 DNA amplification. PMID- 10686099 TI - Crystal structures of mutant monomeric hexokinase I reveal multiple ADP binding sites and conformational changes relevant to allosteric regulation. AB - Hexokinase I, the pacemaker of glycolysis in brain tissue, is composed of two structurally similar halves connected by an alpha-helix. The enzyme dimerizes at elevated protein concentrations in solution and in crystal structures; however, almost all published data reflect the properties of a hexokinase I monomer in solution. Crystal structures of mutant forms of recombinant human hexokinase I, presented here, reveal the enzyme monomer for the first time. The mutant hexokinases bind both glucose 6-phosphate and glucose with high affinity to their N and C-terminal halves, and ADP, also with high affinity, to a site near the N terminus of the polypeptide chain. Exposure of the monomer crystals to ADP in the complete absence of glucose 6-phosphate reveals a second binding site for adenine nucleotides at the putative active site (C-half), with conformational changes extending 15 A to the contact interface between the N and C-halves. The structures reveal distinct conformational states for the C-half and a rigid-body rotation of the N-half, as possible elements of a structure-based mechanism for allosteric regulation of catalysis. PMID- 10686100 TI - Stability of a structural scaffold upon activity transfer: X-ray structure of a three fingers chimeric protein. AB - Fasciculin 2 and toxin alpha proteins belong to the same structural family of three-fingered snake toxins. They act on different targets, but in each case the binding region involves residues from loops I and II. The superimposition of the two structures suggests that these functional regions correspond to structurally distinct zones. Loop I, half of loop II and the C-terminal residue of fasciculin 2 were therefore transferred into the toxin alpha. The inhibition constant of the resulting chimera is only 15-fold lower than that of fasciculin 2, and as expected the potency of binding to the toxin alpha target has been lost. In order to understand the structure-function relationship between the chimera and its "parent" molecules, we solved its structure by X-ray crystallography. The protein crystallized in space group P3(1)21 with a=b=58.5 A, and c=62.3 A. The crystal structure was solved by molecular replacement and refined to 2.1 A resolution. The structure belongs to the three-fingered snake toxin family with a core of four disulphide bridges from which emerge the three loops I, II and III. Superimposition of the chimera on fasciculin 2 or toxin alpha revealed an overall fold intermediate between those of the two parent molecules. The regions corresponding to toxin alpha and to fasciculin 2 retained their respective geometries. In addition, the chimera protein displayed a structural behaviour similar to that of fasciculin 2, i.e. dimerization in the crystal structure of fasciculin 2, and the geometry of the region that binds to acetylcholinesterase. In conclusion, this structure shows that the chimera retains the general structural characteristics of three-fingered toxins, and the structural specificity of the transferred function. PMID- 10686101 TI - Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase alpha-cyclodextrin production. AB - Cyclodextrin glycosyltransferases (CGTase) (EC 2.4.1.19) are extracellular bacterial enzymes that generate cyclodextrins from starch. All known CGTases produce mixtures of alpha, beta, and gamma-cyclodextrins. A maltononaose inhibitor bound to the active site of the CGTase from Bacillus circulans strain 251 revealed sugar binding subsites, distant from the catalytic residues, which have been proposed to be involved in the cyclodextrin size specificity of these enzymes. To probe the importance of these distant substrate binding subsites for the alpha, beta, and gamma-cyclodextrin product ratios of the various CGTases, we have constructed three single and one double mutant, Y89G, Y89D, S146P and Y89D/S146P, using site-directed mutagenesis. The mutations affected the cyclization, coupling; disproportionation and hydrolyzing reactions of the enzyme. The double mutant Y89D/S146P showed a twofold increase in the production of alpha-cyclodextrin from starch. This mutant protein was crystallized and its X ray structure, in a complex with a maltohexaose inhibitor, was determined at 2.4 A resolution. The bound maltohexaose molecule displayed a binding different from the maltononaose inhibitor, allowing rationalization of the observed change in product specificity. Hydrogen bonds (S146) and hydrophobic contacts (Y89) appear to contribute strongly to the size of cyclodextrin products formed and thus to CGTase product specificity. Changes in sugar binding subsites -3 and -7 thus result in mutant proteins with changed cyclodextrin production specificity. PMID- 10686102 TI - Is folding of beta-lactoglobulin non-hierarchic? Intermediate with native-like beta-sheet and non-native alpha-helix. AB - The refolding of beta-lactoglobulin, a beta-barrel protein consisting of beta strands betaA-betaI and one major helix, is unusual because non-native alpha helices are formed at the beginning of the process. We studied the refolding kinetics of bovine beta-lactoglobulin A at pH 3 using the stopped-flow circular dichroism and manual H/(2)H exchange pulse labeling coupled with heteronuclear NMR. The protection pattern from the H/(2)H exchange of the native state indicated the presence of a stable hydrophobic core consisting of betaF, betaG and betaH strands. The protection pattern of the kinetic intermediate obtained about one second after initiating the reaction was compared with that of the native state. In this relatively late kinetic intermediate, which still contains some non-native helical structure, the disulfide-bonded beta-hairpin made up of betaG and betaH strands was formed, but the rest of the molecule was fluctuating, where the non-native alpha-helices may reside. Subsequently, the core beta-sheet extends, accompanied by a further alpha-helix to beta-sheet transition. Thus, the refolding of beta-lactoglobulin exhibits two elements: the critical role of the core beta-sheet is consistent with the hierarchic mechanism, whereas the alpha helix to beta-sheet transition suggests the non-hierarchic mechanism. PMID- 10686103 TI - Thermodynamics of DNA binding and condensation: isothermal titration calorimetry and electrostatic mechanism. AB - The thermodynamics of binding of the trivalent cations cobalt hexammine and spermidine to plasmid DNA was studied by isothermal titration calorimetry. Two stages were observed in the course of titration, the first attributed to cation binding and the second to DNA condensation. A standard calorimetric data analysis was extended by applying an electrostatic binding model, which accounted for most of the observed data. Both the binding and condensation reactions were entropically driven (TDeltaS approximately +10 kcal/mol cation) and enthalpically opposed (DeltaH approximately +1 kcal/mol cation). As predicted from their relative sizes, the binding constants of the cations were indistinguishable, but cobalt hexammine had a much greater DNA condensing capacity because it is more compact than spermidine. The dependence of both the free energy of cobalt hexammine binding and the critical cobalt hexammine concentration for DNA condensation on temperature and monovalent cation concentration followed the electrostatic model quite precisely. The heat capacity changes of both stages were positive, perhaps reflecting both the temperature dependence of the dielectric constant of water and the burial of polar surfaces. DNA condensation occurred when about 67 % of the DNA phosphate charge was neutralized by cobalt hexammine and 87 % by spermidine. During condensation, the remaining DNA charge was neutralized. PMID- 10686104 TI - Backbone dynamics of a cbEGF domain pair in the presence of calcium. AB - Calcium binding (cb) epidermal growth factor-like (EGF) domains are found in a wide variety of extracellular proteins with diverse functions. In several proteins, including the fibrillins (1 and 2), the low-density lipoprotein receptor, the Notch receptor and related molecules, these domains are organised as multiple tandem repeats. The functional importance of calcium-binding by EGF domains has been underscored by the identification of missense mutations associated with defective calcium-binding, which have been linked to human diseases. Here, we present (15)N backbone relaxation data for a pair of cbEGF domains from fibrillin-1, the defective protein in the Marfan syndrome. The data were best fit using a symmetric top model, confirming the extended conformation of the cbEGF domain pair. Our data demonstrate that calcium plays a key role in stabilising the rigidity of the domain pair on the pico- to millisecond time scale. Strikingly, the most dynamically stable region of the construct is centred about the domain interface. These results provide important insight into the properties of intact fibrillin-1, the consequences of Marfan syndrome causing mutations, and the ultrastructure of fibrillins and other extracellular matrix proteins. PMID- 10686105 TI - The structural basis for enhanced stability and reduced DNA binding seen in engineered second-generation Cro monomers and dimers. AB - It was previously shown that the Cro repressor from phage lambda, which is a dimer, can be converted into a stable monomer by a five-amino acid insertion. Phe58 is the key residue involved in this transition, switching from interactions which stabilize the dimer to those which stabilize the monomer. Structural studies, however, suggested that Phe58 did not penetrate into the core of the monomer as well as it did into the native dimer. This was strongly supported by the finding that certain core-repacking mutations, including in particular, Phe58 ->Trp, increased the stability of the monomer. Unexpectedly, the same substitution also increased the stability of the native dimer. At the same time it decreased the affinity of the dimer for operator DNA. Here we describe the crystal structures of the Cro F58W mutant, both as the monomer and as the dimer. The F58W monomer crystallized in a form different from that of the original monomer. In contrast to that structure, which resembled the DNA-bound form of Cro, the F58W monomer is closer in structure to wild-type (i.e. non-bound) Cro. The F58W dimer also crystallizes in a form different from the native dimer but has a remarkably similar overall structure which tends to confirm the large changes in conformation of Cro on binding DNA. Introduction of Trp58 perturbs the position occupied by the side-chain of Arg38, a DNA-contact residue, providing a structural explanation for the reduction in DNA-binding affinity. The improved thermal stability is seen to be due to the enhanced solvent transfer free energy of Trp58 relative to Phe58, supplemented in the dimer structure, although not the monomer, by a reduction in volume of internal cavities. PMID- 10686106 TI - Molecular dynamics simulations of a beta-hairpin fragment of protein G: balance between side-chain and backbone forces. AB - How is the native structure encoded in the amino acid sequence? For the traditional backbone centric view, the dominant forces are hydrogen bonds (backbone) and phi-psi propensity. The role of hydrophobicity is non-specific. For the side-chain centric view, the dominant force of protein folding is hydrophobicity. In order to understand the balance between backbone and side chain forces, we have studied the contributions of three components of a beta hairpin peptide: turn, backbone hydrogen bonding and side-chain interactions, of a 16-residue fragment of protein G. The peptide folds rapidly and cooperatively to a conformation with a defined secondary structure and a packed hydrophobic cluster of aromatic side-chains. Our strategy is to observe the structural stability of the beta-hairpin under systematic perturbations of the turn region, backbone hydrogen bonds and the hydrophobic core formed by the side-chains, respectively. In our molecular dynamics simulations, the peptides are solvated. with explicit water molecules, and an all-atom force field (CFF91) is used. Starting from the original peptide (G41EWTYDDATKTFTVTE56), we carried out the following MD simulations. (1) unfolding at 350 K; (2) forcing the distance between the C(alpha) atoms of ASP47 and LYS50 to be 8 A; (3) deleting two turn residues (Ala48 and Thr49) to form a beta-sheet complex of two short peptides, GEWTYDD and KTFTVTE; (4) four hydrophobic residues (W43, Y45, F52 and T53) are replaced by a glycine residue step-by-step; and (5) most importantly, four amide hydrogen atoms (T44, D46, T53, and T55, which are crucial for backbone hydrogen bonding), are substituted by fluorine atoms. The fluorination not only makes it impossible to form attractive hydrogen bonding between the two beta-hairpin strands, but also introduces a repulsive force between the two strands due to the negative charges on the fluorine and oxygen atoms. Throughout all simulations, we observe that backbone hydrogen bonds are very sensitive to the perturbations and are easily broken. In contrast, the hydrophobic core survives most perturbations. In the decisive test of fluorination, the fluorinated peptide remains folded under our simulation conditions (5 ns, 278 K). Hydrophobic interactions keep the peptide folded, even with a repulsive force between the beta-strands. Thus, our results strongly support a side-chain centric view for protein folding. PMID- 10686107 TI - Preformed secondary structure drives the association reaction of GCN4-p1, a model coiled-coil system. AB - The structure of the transition state for the rate-limiting step in the folding and association of the homodimeric coiled-coil peptide GCN4-p1, was probed by mutational analysis. A series of quadruple amino acid replacements that spanned the helix propensity scale were made at the four external f positions in the heptad repeat. Equilibrium and kinetic circular dichroism studies demonstrate that both the stability and the unfolding and refolding rate constants vary with helix propensity but also reflect interactions of the altered side-chains with their local environments. Pairwise replacements and fragment studies show that the two C-terminal heptads are the likely source of the nucleating helices. Helix helix recognition between preformed elements of secondary structure plays an important role in this fundamental folding reaction. PMID- 10686108 TI - Three-dimensional structure of a presynaptic neurotoxic phospholipase A2 from Daboia russelli pulchella at 2.4 A resolution. AB - The phospholipase A(2 )from Daboia russelli pulchella (DPLA(2)) is the only known member of subclass II of group IIA. The three-dimensional structure of this presynaptic neurotoxic DPLA(2) enzyme has been determined at 2.4 A resolution. The structure was determined by the molecular replacement method using the model Crotalus atrox, and refined using X-PLOR to a final R-factor of 18.8 % for all data in the resolution range 20.0 A-2.4 A. The final refined model comprises 1888 atoms from two crystallographically independent protein molecules and 160 water oxygen atoms. The overall folding of DPLA(2), with three long helices and two short antiparallel beta-strands is grossly similar to those observed for other PLA(2)s. In the present structure, the calcium binding site is empty but the conformation of the calcium binding loop is similar to those observed in the calcium bound states. Two spatially adjacent regions of residues 55-61 (a typical beta-turn I) and 83-94 (a well defined loop) are remarkably different in conformation, electrostatic characteristics and inter-segmental interactions from those found in non-neurotoxic PLA(2)s. Yet another striking structural feature in DPLA(2 )pertains to the stretch of residues 53-77, which has a series of positively charged residues protruding outwardly. The above segment is presumed to be involved in the anticoagulant activity. A unique hydrophobic patch including residues Leu17, Ala18, Ile19, Pro20, Phe106 and Leu110 is found on the surface together with an equally emphatic region of -OH groups containing residues such as Ser21, Tyr22, Ser23, Ser24, Tyr25 and Tyr28. The interactions between two molecules of DPLA(2) in the asymmetric unit are remarkably different from those observed in the standard dimers and trimers of PLA(2)s, leaving the enzyme's active site fully exposed for enzyme-substrate reactions, it makes this structure one of the most favourable examples for structure-based drug design through soaking experiments. PMID- 10686109 TI - Luminescence control in the marine bacterium Vibrio fischeri: An analysis of the dynamics of lux regulation. AB - A mathematical model has been developed based on the fundamental properties of the control system formed by the lux genes and their products in Vibrio fischeri. The model clearly demonstrates how the components of this system work together to create two, stable metabolic states corresponding to the expression of the luminescent and non-luminescent phenotypes. It is demonstrated how the cell can "switch" between these steady states due to changes in parameters describing metabolic processes and the extracellular concentration of the signal molecule N 3-oxohexanoyl-l-homoserine lactone. In addition, it is shown how these parameters influence how sensitive the switch mechanism is to cellular LuxR and N-3 oxohexanoyl-l-homoserine lactone and complex concentration. While these properties could lead to the collective phenomenon known as quorum sensing, the model also predicts that under certain metabolic circumstances, basal expression of the lux genes could cause a cell to luminesce in the absence of extracellular signal molecule. Finally, the model developed in this study provides a basis for analysing the impact of other levels of control upon lux regulation. PMID- 10686110 TI - Characterization of novel proteins based on known protein structures. AB - The genome sciences face the challenge to characterize structure and function of a vast number of novel genes. Sequence search techniques are used to infer functional and structural information from similarities to experimentally characterized genes or proteins. The persistent goal is to refine these techniques and to develop alternative and complementary methods to increase the range of reliable inference.Here, we focus on the structural and functional assignments that can be inferred from the known three-dimensional structures of proteins. The study uses all structures in the Protein Data Bank that were known by the end of 1997. The protein structures released in 1998 were then characterized in terms of functional and structural similarity to the previously known structures, yielding an estimate of the maximum amount of information on novel protein sequences that can be obtained from inference techniques. The 147 globular proteins corresponding to 196 domains released in 1998 have no clear sequence similarity to previously known structures. However, 75 % of the domains have extensive structure similarity to previously known folds, and most importantly, in two out of three cases similarity in structure coincides with related function. In view of this analysis, full utilization of existing structure data bases would provide information for many new targets even if the relationship is not accessible from sequence information alone. Currently, the most sophisticated techniques detect of the order of one-third of these relationships. PMID- 10686111 TI - A motif for quinone binding sites in respiratory and photosynthetic systems. AB - Many of the membrane-bound protein complexes of respiratory and photosynthetic systems are reactive with quinones. To date, no clear structural relationship between sites that bind quinone has been defined, apart from that in the homologous family of "type II" photosynthetic reaction centres. We show here that a structural element containing a weak sequence motif is common to the Q(A) and Q(B) sites of bacterial reaction centres and the Q(i) site of the mitochondrial bc(1) complex. Analyses of sequence databases indicate that this element may also be present in the PsaA/B subunits of photosystem I, in the ND4 and ND5 subunits of complex I and, possibly, in the mitochondrial alternative quinol oxidase. This represents a first step in the structural classification of quinone binding sites. PMID- 10686112 TI - Time perception and motor timing: a common cortical and subcortical basis revealed by fMRI. AB - Though it is well known that humans perceive the temporal features of the environment incessantly, the brain mechanisms underlying temporal processing are relatively unexplored. Functional magnetic resonance imaging was used in this study to identify brain activations during sustained perceptual analysis of auditorally and visually presented temporal patterns (rhythms). Our findings show that the neural network supporting time perception involves the same brain areas that are responsible for the temporal planning and coordination of movements. These results indicate that time perception and motor timing rely on similar cerebral structures. PMID- 10686113 TI - Cocaine activation discriminates dopaminergic projections by temporal response: an fMRI study in Rat. AB - We applied a sensitive new functional magnetic resonance imaging technique to identify the pattern and determinants of cocaine-induced brain activation in drug naive rats. At doses greater than 0.1 mg/kg iv, cocaine produced robust activation throughout cortex with the largest magnitude increase in frontal neocortex. Additionally, we detected selective activation within dopamine innervated subcortical regions including dorsomedial and ventrolateral striatum, nucleus accumbens region, and dorsal thalamus. Although dose response was similar among activated regions, temporal response differentiated regions along distinct anatomical boundaries with basal ganglia and limbic cortical structures, reaching maximum activation later than frontal neocortex. Pharmacological specificity was demonstrated by blocking cocaine-induced activation with SCH-23390, a selective D1 antagonist. Our data demonstrate the utility of fMRI to identify spatiotemporal patterns of cocaine-induced brain activation and implicate D1 dopaminergic mechanisms in acute cocaine action. PMID- 10686114 TI - Detection of consistently task-related activations in fMRI data with hybrid independent component analysis. AB - fMRI data are commonly analyzed by testing the time course from each voxel against specific hypothesized waveforms, despite the fact that many components of fMRI signals are difficult to specify explicitly. In contrast, purely data-driven techniques, by focusing on the intrinsic structure of the data, lack a direct means to test hypotheses of interest to the examiner. Between these two extremes, there is a role for hybrid methods that use powerful data-driven techniques to fully characterize the data, but also use some a priori hypotheses to guide the analysis. Here we describe such a hybrid technique, HYBICA, which uses the initial characterization of the fMRI data from Independent Component Analysis and allows the experimenter to sequentially combine assumed task-related components so that one can gracefully navigate from a fully data-derived approach to a fully hypothesis-driven approach. We describe the results of testing the method with two artificial and two real data sets. A metric based on the diagnostic Predicted Sum of Squares statistic was used to select the best number of spatially independent components to combine and utilize in a standard regressional framework. The proposed metric provided an objective method to determine whether a more data-driven or a more hypothesis-driven approach was appropriate, depending on the degree of mismatch between the hypothesized reference function and the features in the data. HYBICA provides a robust way to combine the data derived independent components into a data-derived activation waveform and suitable confounds so that standard statistical analysis can be performed. PMID- 10686115 TI - Passive and active recognition of one's own face. AB - Facial identity recognition has been studied mainly with explicit discrimination requirement and faces of social figures in previous human brain imaging studies. We performed a PET activation study with normal volunteers in facial identity recognition tasks using the subject's own face as visual stimulus. Three tasks were designed so that the activation of the visual representation of the face and the effect of sustained attention to the representation could be separately examined: a control-face recognition task (C), a passive own-face recognition task (no explicit discrimination was required) (P), and an active own-face recognition task (explicit discrimination was required) (A). Increased skin conductance responses during recognition of own face were seen in both task P and task A, suggesting the occurrence of psychophysiological changes during recognition of one's own face. The left fusiform gyrus, the right supramarginal gyrus, the left putamen, and the right hypothalamus were activated in tasks P and A compared with task C. The left fusiform gyrus and the right supramarginal gyrus are considered to be involved in the representation of one's own face. The activation in the right supramarginal gyrus may be associated with the representation of one's own face as a part of one's own body. The prefrontal cortices, the right anterior cingulate, the right presupplementary motor area, and the left insula were specifically activated during task A compared with tasks C and P, indicating that these regions may be involved in the sustained attention to the representation of one's own face. PMID- 10686116 TI - Characterization and correction of interpolation effects in the realignment of fMRI time series. AB - Subject motion in functional magnetic resonance imaging (fMRI) studies can be accurately estimated using realignment algorithms. However, residual changes in signal intensity arising from motion have been identified in the data even after realignment of the image time series. The nature of these artifacts is characterized using simulated displacements of an fMRI image and is attributed to interpolation errors introduced by the resampling inherent within realignment. A correction scheme that uses a periodic function of the estimated displacements to remove interpolation errors from the image time series on a voxel-by-voxel basis is proposed. The artifacts are investigated using a brain phantom to avoid physiological confounds. Small- and large-scale systematic displacements show that the artifacts have the same form as revealed by the simulated displacements. A randomly displaced phantom and a human subject are used to demonstrate that interpolation errors are minimized using the correction. PMID- 10686117 TI - Real-time monitoring of eye movements using infrared video-oculography during functional magnetic resonance imaging of the frontal eye fields. AB - Monitoring eye movements is a critical aspect of experimental design for studies of spatial attention and visual perception. However, obtaining online eye movement recordings has been technologically difficult during functional magnetic resonance (MR) imaging studies. Previous approaches to monitoring eye movements either have distorted the MR images or have shown MR-related interference in the recordings. We report a technique using long-range infrared video-oculography to record eye movements without causing artifacts in the MR images. Analysis of the MR signal from a phantom obtained with the eye-movement equipment turned on or off confirmed the absence of significant additional noise in the MR time series. Eye movements of three subjects were monitored while they performed tasks of covert and overt shifts of spatial attention. Activation of the frontal eye fields during the covert task was seen even when the eye-movement recordings demonstrated no significant difference in saccadic eye movements between the baseline and the active conditions. PMID- 10686118 TI - Brodmann's areas 17 and 18 brought into stereotaxic space-where and how variable? AB - Studies on structural-functional associations in the visual system require precise information on the location and variability of Brodmann's areas 17 and 18. Usually, these studies are based on the Talairach atlas, which does not rely on cytoarchitectonic observations, but on comparisons of macroscopic features in the Talairach brain and Brodmann's drawing. In addition, in this atlas are found only the approximate positions of cytoarchitectonic areas and not the exact borders. We have cytoarchitectonically mapped both areas in 10 human brains and marked their borders in corresponding computerized images. Borders were defined on the basis of quantitative cytoarchitecture and multivariate statistics. In addition to borders of areas 17 and 18, subparcellations within both areas were found. The cytoarchitectonically defined areas were 3-D reconstructed and transferred into the stereotaxic space of the standard reference brain. Surface rendering of the brains revealed high individual variability in size and shape of the areas and in the relationship to the free surface and sulci. Ranges and centers of gravity of both areas were calculated in Talairach coordinates. The positions of areas 17 and 18 in the stereotaxic space differed between the hemispheres. Both areas reached significantly more caudal and medial positions on the left than on the right. Probability maps were created in which the degree of overlap in each stereotaxic position was quantified. These maps of areas 17 and 18 are the first of their kind and contain precise stereotaxic information on both interhemispheric and interindividual differences. PMID- 10686119 TI - Effects of environmental enrichment on rate of contextual processing and discriminative ability in adult rats. AB - The effect of environmental enrichment on conditioned freezing to contextual cues in adult Sprague-Dawley rats was examined. The freezing of both enriched-and standard-reared rats increased with the time spent in the chamber prior to shock. Both groups of rats showed equally low levels of contextual conditioning following a preshock period of 4 s and equally high levels following a 120-s preshock period. However, following a preshock period of 16 s enriched rats displayed more contextual conditioning than standard rats. That is, enriched rats appeared to process contextual information faster than their standard-reared counterparts. Enriched- reared rats also showed a greater ability to discriminate between the conditioning context and a similar but distinctive context. Hence, in addition to forming a representation of the context in memory more rapidly than standard-reared rats, enriched-reared rats also appear to form a more complex representation. PMID- 10686120 TI - Time-dependent impairment of inhibitory avoidance retention in rats by posttraining infusion of a mitogen-activated protein kinase kinase inhibitor into cortical and limbic structures. AB - Mitogen-activated protein kinase (MAPK) is abundantly expressed in postmitotic neurons of the developed nervous system. MAPK is activated and required for induction of long-term potentiation (LTP) in the CA1 area of the hippocampus, which is blocked by the specific inhibitor of the MAPK kinase, PD 098059. Recently it was demonstrated that MAPK is activated in the hippocampus after training and is necessary for contextual fear conditioning learning. The present work tests the role of the MAPK cascade in step-down inhibitory avoidance (IA) retention. PD 098059 (50 microM) was bilaterally injected (0.5 microl/side) into the CA1 region of the dorsal hippocampus or entorhinal cortex at 0, 90, 180, or 360 min, or into the amygdala or parietal cortex at 0, 180, or 360 min after IA training in rats using a 0.4-mA foot shock. Retention testing was carried out 24 h after training. PD 098059 impaired retention when injected into the dorsal hippocampus at 180 min, but not 0, 90, and 360 min after training. When infused into the entorhinal cortex, PD 098059 was amnestic at 0 and 180 min, but not at 90 and 360 min after training. The MAPKK inhibitor also impairs IA retention when infused into the parietal cortex immediately after training, but not at 180 or 360 min. Infusions performed into amygdala were amnestic at 180 min, but not at 0 and 360 min after training. Our results suggest a time-dependent involvement of the MAPK cascade in the posttraining memory processing of IA; the time dependency is different in the hippocampus, amygdala, entorhinal cortex, or parietal cortex of rats. PMID- 10686121 TI - Injections of tacrine and scopolamine into the nucleus accumbens: opposing effects of immediate vs delayed posttrial treatment on memory of an open field. AB - Using the paradigm of habituation learning in the open field, we tested the effects of microinjections of the nonspecific acetylcholine-esterase inhibitor tacrine (0.1, 1.0, 10.0 micrograms), and the muscarinic receptor antagonist scopolamine (0.1, 1.0, 10.0 micrograms) into the core of the nucleus accumbens. When injected immediately after the first exposure to the open field (posttrial), tacrine dose-dependently enhanced habituation of rearing behavior during the test on the following day, indicating a facilitation of memory. In contrast, scopolamine impaired habituation of rearing behavior at the two lower doses, but not at the highest dose. When scopolamine or tacrine (10.0 micrograms) was injected with a delay of 5 h after the learning trial, both drugs impaired habituation of rearing on the following day. The effects on locomotor activity differed from those on rearing behavior. Here, habituation on Day 2 was observed only in those animals which had received posttrial injections of vehicle or 10 micrograms of tacrine on the day before, whereas in animals which had received the two lower doses of tacrine, locomotor activity on Day 2 was not significantly decreased. In animals with posttrial treatment of scopolamine, locomotor activity on Day 2 was even enhanced, especially with the lower doses. No such effects were observed when scopolamine or tacrine (10.0 micrograms each) was injected with a delay of 5 h after the learning trial. These results show that cholinergic manipulations aimed at the nucleus accumbens can have substantial effects in this posttrial memory paradigm, which depend on drug, dose, and time of injection, and the specific kind of behavioral measure analyzed. Among others, the findings are discussed with respect to the role of muscarinic and nicotinergic cholinergic mechanisms in the nucleus accumbens on cognitive functions. They may be relevant, for example, for understanding the psychopathology of Alzheimer's disease, since the nucleus accumbens is one of the sites where cholinergic neurons are lost in this neurodegenerative disease. PMID- 10686122 TI - Genetic differences in response to novelty and spatial memory using a two-trial recognition task in mice. AB - A two-trial memory task, based on a free-choice exploration paradigm in a Y-maze, was previously developed to study recognition processes in Sprague-Dawley rats. Because this paradigm avoids the use of electric shock or deprivation that may have nonspecific effects and does not require learning of a rule, it may be particularly useful for studying memory in mice. Four inbred strains (Balb/cByJ, DBA/2J, C57BL/6J, and SJL/J), an F1 hybrid (C57BL/6 x SJL/J), and one outbred strain (CD1) were used to validate this task in mice and to characterize a strain distribution in response to novelty and working memory. Exploration was measured with a short (2 min) intertrial interval (ITI) between acquisition and retrieval, while memory was examined with longer intervals (30 min, 1 h, and 2 h). A study of the time course of the response to novelty revealed varying degrees of preference and/or habituation to novelty among the different strains, with CD1 exhibiting a very high response to novelty and others showing lower (C57 x SJL hybrids) to complete absence (SJL) of exploration of novelty. Memory span, assessed with increasing ITIs, varied widely among strains from 30 min (C57 x SJL hybrids) to at least 2 h (C57 and BALB). Such demonstrated sensitivity to a wide range of behavioral phenotypes supports the use of this spatial memory task as an effective tool for the study of genetic influences on the response to novelty and recognition processes in mice. PMID- 10686123 TI - Enhanced visuospatial memory following intracerebroventricular administration of nerve growth factor. AB - The present work assessed the effects of intracerebroventricular injections of rh recombined human nerve growth factor (rh NGF) (5 micrograms/2.5 microl) at postnatal days 12 and 13 upon the development of spatial learning capacities. The treated rats were trained at the age of 22 days to escape onto an invisible platform at a fixed position in space in a Morris navigation task. For half of the subjects, the training position was also cued, a procedure aimed at facilitating escape and at reducing attention to the distant spatial cues. Later, at the age of 6 months, all the rats were trained in a radial-arm maze task. Treatment effects were found in both immature and adult rats. The injection of NGF improved the performance in the Morris navigation task in both training conditions. There was a significant reduction in the escape latency and an increased bias toward the training platform quadrant during probe trials. The most consistent effect was the precocious development of an adult-like spatial memory. In the radial-arm maze, the NGF-treated rats made significantly fewer reentries than vehicle rats and this effect was particularly marked in the treated female rats. Taken together, these experiments reveal that the development and the maintenance of an accurate spatial representation are tightly related to the development of brain structures facilitated by the action of NGF. Moreover, these experiments demonstrate that an acute pharmacological treatment that leads to a transient modification in the choline acetyltransferase activity can induce a behavioral change long after the treatment. PMID- 10686124 TI - Effects of selective perirhinal and postrhinal lesions on acquisition and retention of a visual discrimination task in rats. AB - The hippocampal region along with rhinal structures seems to support learning and memory in an important manner. Structures adjacent to the rhinal fissure in the rat have recently been suggested to be divided into the perirhinal and postrhinal cortices. Some effects of perirhinal lesions on cognitive processing are known, whereas effects of postrhinal lesions appear to be unknown. The purpose of the present study was to examine the relative effects of perirhinal and postrhinal lesions in a three-choice visual discrimination test. The results show that both types of lesions impaired acquisition of the task, but only perirhinal lesions impeded subsequent retention. Because the initial phase of acquisition was unaffected by both lesions, it is suggested that the deficits observed may be of mnemonic nature. The apparent differential involvement of the perirhinal cortex and postrhinal cortex in cognition may be associated with differences in anatomical connectivity among these structures. PMID- 10686125 TI - Epinephrine fails to enhance performance of food-deprived rats on a delayed spontaneous alternation task. AB - Increases in blood glucose levels after epinephrine injection appear to contribute to the hormone's effects on learning and memory. The present experiment evaluated whether epinephrine-induced enhancement of spontaneous alternation performance would be attenuated in fasted rats that had blunted increases in circulating glucose levels after injections of epinephrine. Rats deprived of food for 24 h prior to injection of epinephrine exhibited significant attenuation of the increase in blood glucose levels seen in fed rats. When the rats were tested on a delayed spontaneous alternation task, epinephrine enhanced performance in fed rats but not in rats deprived of food for 24 h. These findings are consistent with the view that hyperglycemia subsequent to epinephrine injections contributes to the memory-enhancing effects of epinephrine. PMID- 10686126 TI - Evidence for altered Fragile-X mental retardation protein expression in response to behavioral stimulation. AB - The Fragile-X mental retardation protein, the protein absent in Fragile-X syndrome, is synthesized near synapses upon neurotransmitter activation. Humans and mice lacking this protein exhibit abnormal dendritic spine lengths and numbers. Here we investigated Fragile-X protein levels in animals exposed to behavioral paradigms that induce neuronal morphological change. Fragile-X protein immunoreactivity was examined in visual cortices of rats reared in a complex environment for 10 or 20 days, motor cortices of rats trained on motor-skill tasks for 3 or 7 days, and either visual or motor cortices of inactive controls. Rats exposed to a complex environment for 20 days or trained for 7 days on motor skill tasks exhibited increased Fragile-X protein immunoreactivity in visual or motor cortices, respectively. These results provide the first evidence for a behaviorally induced alteration of Fragile-X protein expression and are compatible with previous findings suggesting synaptic regulation of its expression. These results also strengthen the association of Fragile-X mental retardation protein expression with the alteration of synaptic structure. PMID- 10686127 TI - Shuttle-box avoidance learning in mice: improvement by glucose combined with stimulant drugs. AB - Glucose was tested alone or in combination with two stimulant drugs, amphetamine and nicotine, in mice of the CD-1 strain subjected to five daily shuttle-box training sessions. Pretraining intraperitoneal administration of glucose (50 or 100 mg/kg) had no effect, while amphetamine and nicotine, given alone, significantly improved avoidance acquisition at a dose of 0.5 mg/kg, but not 0.025 mg/kg. Significant improvement of avoidance learning was also produced by a combination of glucose with the lower dose of amphetamine or nicotine. This enhancing action, produced by a combination of glucose and stimulant drugs, at doses ineffective by themselves, might be due to a concomitant cholinergic and dopaminergic activation, induced by glucose and stimulant drugs, respectively. PMID- 10686128 TI - The genes for benzene catabolism in Pseudomonas putida ML2 are flanked by two copies of the insertion element IS1489, forming a class-I-type catabolic transposon, Tn5542. AB - Two directly repeated sequences of the IS elements IS1489v1 and IS1489v2 flank the benzene dioxygenase (bedC1C2BA) and the cis-benzene dihydrodiol dehydrogenase (bedD) genes on the catabolic plasmid pHMT112 in Pseudomonas putida ML2, forming a Class-I-type composite transposon, Tn5542. Both IS1489v1 and IS1489v2 contain an identical 1371-bp open reading frame, tnpA, that is preceded by a possible ribosome binding site. The tnpA gene of IS1489v1 is bound by a pair of 40-bp imperfect inverted repeats while that of IS1489v2 is flanked only by the left inverted repeat. The tnpA gene codes for a putative 53-kDa polypeptide of 456 amino acids bearing similarity to transposases encoded on IS elements of P. alcaligenes, P. aeruginosa, P. stutzeri, and Serratia marcescens. The basic nature of the putative TnpA protein with a deduced pI of 8.93 is typical of IS encoded transposases. Similar to other IS elements, an outward facing promoter was detected at the right end of IS1489v1. Experiments involving the suicide vector, pKNG101, failed to show transposition of Tn5542. PMID- 10686129 TI - Nucleotide sequence and analysis of plasmid pMD136 from Pediococcus pentosaceus FBB61 (ATCC43200) involved in pediocin A production. AB - The complete sequence of the 19515-bp plasmid pMD136 from Pediococcus pentosaceus FBB61 (ATCC43200) has been determined. This plasmid is involved in Pediocin A production, a bacteriocin active against a wide range of gram-positive bacteria. It appears to replicate via a theta mechanism, with structures closely related to those of many lactococcal plasmids. Genes homologous to mobilization functions are also present, which are similar in sequence and arrangement to mobA, mobB, and mobC of some staphylococcal plasmids, although the last one contains a deletion in its central part. The region involved in bacteriocin activity has been limited to a 9.4-kb fragment, containing 10 open reading frames organized in a single operon. Since Pediocin A has a molecular weight of about 80 kDa (Piva and Headon, Microbiology, 140, 697-702, 1994), and a gene long enough to encode it is not present in pMD136, it is proposed that genes residing on the plasmid are responsible for the regulation of bacteriocinogenic activity. Gene arrangement and sequence homologies suggest the presence of a two-component-like regulatory mechanism. PMID- 10686131 TI - Nucleotide sequence analysis of the lactococcal EPS plasmid pNZ4000. AB - The complete 42180-bp nucleotide sequence of the mobilization plasmid pNZ4000, coding for exopolysaccharide (EPS) production in Lactococcus lactis, was determined. This plasmid contains a region involved in EPS biosynthesis, four functional replicons, a region containing mobilization genes, and three origin of transfer (oriT) sequences. Sequences identical to these oriT sequences were also found on two other lactococcal plasmids and a plasmid from Lactobacillus helveticus. Several complete and partial IS elements were identified on pNZ4000, including iso-ISS1, iso-IS946, and iso-IS982 sequences. Furthermore, pNZ4000 contains a gene cluster that may encode a cobalt transport system and a gene that encodes a CorA homologue which may function as a magnesium transporter. PMID- 10686130 TI - Coupling sequences flanking Tn916 do not determine the affinity of binding of integrase to the transposon ends and adjacent bacterial DNA. AB - Coupling sequences are the 6 bp flanking the conjugative transposon Tn916 and are thought to play a role in determining the frequency of conjugative transposition. The affinity of binding of a chimeric protein, which consisted of maltose binding protein fused to the carboxy-terminal DNA binding domain of Tn916 integrase (Int), to different double-stranded oligonucleotide substrates containing coupling sequences associated with high- and low-frequency conjugative transposition was measured using a competition binding assay. The relative affinity of the chimeric protein was unaffected by the nature of the coupling sequences tested. The same results were obtained when the coupling sequences were placed in a different surrounding sequence context. It therefore appears that the effects of different coupling sequences on the frequency of conjugative transposition are not due simply to differences in Int binding. PMID- 10686132 TI - Telomere sequences attached to nuclearly migrated yeast linear plasmid. AB - The yeast linear plasmid pCLU1, derived from pGKL1, has terminal proteins (TPs) covalently attached at the 5' ends of inverted terminal repeats (ITRs) and replicates in the cytoplasm, presumably using the TP as a primer for DNA synthesis. In Saccharomyces cerevisiae, under certain conditions, pCLU1 migrated into the nucleus and replicated in either linear or circular form. The linear form plasmid lacked TPs; instead it carried host-telomere repeats at the ITR ends. The present study showed that (1) the added telomere was primarily composed of the repeated tracts of TGTGTGGGTGTGG, which was complementary to the RNA template of yeast telomerase, (2) the telomeric addition occurred at the very end of the ITRs, and (3) the sequence composition of the added telomeres was diverse among individual plasmids, but symmetrically identical at both ends of each plasmid. A similar mode of telomere addition was also observed in cells defective in the RAD52 gene. PMID- 10686133 TI - Complete DNA sequence and analysis of an emerging cryptic plasmid isolated from Yersinia pestis. AB - A 6-kb cryptic plasmid (pYC; 5919 bp) has been recovered from Yersinia pestis isolates originating from regions of Yunnan province in China. The sequence of pYC was determined, and analysis of the sequence has revealed that two of the plasmid DNA regions (ORFs 10 and 11) are similar to the DinJ1 and DinJ2 gene products encoded by Escherichia coli chromosomal DNA. This plasmid is increasingly harbored by Y. pestis isolates recovered from a domestic rodent cycle in the southern regions of the province. Further studies will determine the origin and function of pYC. PMID- 10686134 TI - The pilL and pilN genes of IncI1 plasmids R64 and ColIb-P9 encode outer membrane lipoproteins responsible for thin pilus biogenesis. AB - The predicted amino acid sequences of the pilL and pilN genes, required for the thin pilus formation of IncI1 plasmids R64 and ColIb-P9, contain N-terminal lipoprotein signal peptide motifs. The pilL and pilN products were labeled with [(3)H]palmitic acid as 38- and 57-kDa proteins, respectively, indicating that they are lipoproteins. Both PilL and PilN were localized to the outer membrane. PMID- 10686135 TI - Detection, cloning, and sequence analysis of an indigenous plasmid from cellulolytic clostridial strain MCF1. AB - Nucleotide sequence analysis of a 2451-bp plasmid (pMCF1) from a cellulolytic Clostridium revealed that the protein specified by the largest open reading frame (ORF1) was homologous to RepB of Clostridium butyricum plasmid pCB101. The data suggest that pMCF1 belongs to the pC194 family of rolling-circle replicating plasmids and the ORF1 protein functions as its replication protein. PMID- 10686136 TI - Complete nucleotide sequence and characterization of pSNA1 from pimaricin producing Streptomyces natalensis that replicates by a rolling circle mechanism. AB - A cryptic plasmid, pSNA1, has been identified in the pimaricin-producing Streptomyces natalensis strain ATCC 27448. pSNA1 has been mapped with restriction endonucleases and its complete nucleotide sequence was determined. The circular DNA molecule is 9367 bp in length and has a 71.3% G+C content. Its estimated copy number is 30. Analysis of the sequence and codon preferences indicated that pSNA1 contains seven open reading frames [encoding peptides larger than 90 amino acid (aa) residues], ORF 1 to ORF 7, located on both strands of pSNA1. ORF 3 codes for a protein (476 aa) that shows high sequence similarity to replication-associated proteins in Streptomyces plasmids known to replicate via the rolling circle mechanism. Accumulation of single-strand intermediates further indicates that pSNA1 replicates via the rolling circle replication model. ORF 1 encodes a polypeptide of 246 aa that shares homology with KorA proteins encoded by other streptomycete plasmids. ORF 4 (SpdA) codes for a protein (161 aa) possibly involved in intramycelial plasmid transfer. Protein encoded by ORF 2 (309 aa) shares homology with a Streptomyces protein (SpdB2) also involved in plasmid spreading. PMID- 10686137 TI - Strand switching during rolling circle replication of plasmid-like DNA circles in the mitochondria of the higher plant Chenopodium album (L.). AB - The structure of sigma-like mitochondrial DNA molecules prepared from suspension cultured cells of Chenopodium album (L.) was studied by electron microscopy. These molecules were highly variable in size, ranging from about 1 to 104 kb, and had single- and double-stranded regions typical for rolling circle replicating intermediates. Partial denaturation studies confirmed that these structures constitute rolling circles. Close inspection of the circle-tail junctions of the replication fork at high magnification suggests that in circles with a double stranded tail, both strands of the tail seem to be covalently attached to the circle in about 27% of the molecules. This observation can be explained by a phenomenon called strand switching or strand splippage during rolling circle replication, similar to a mechanism proposed for bacterial replicons or in vitro replicating constructs harboring bacteriophage T4 replication origins. PMID- 10686138 TI - Transfer of conjugative plasmids and bacteriophage lambda occurs in the presence of antibiotics that prevent de novo gene expression. AB - Plasmids transferred between bacteria prevented from expressing genes by the presence of bacteriostatic antibiotics. Whereas it has long been known that de novo gene expression is not required in donor cells for conjugation, the observations reported here extend the autonomy of plasmid transfer to the early events of establishment in recipients. In addition, this phenomenon was extended to bacteriophage lambda. PMID- 10686139 TI - Possible benefits of kalilo plasmids to their Neurospora hosts. AB - Neurospora mitochondrial plasmids are ubiquitous in natural populations, yet many of them are lethal to their host strains or seem to impose a molecular genetic load. Five pairs of strains of Neurospora tetrasperma and N. crassa with and without kalilo-like plasmids were tested under a variety of situations. The purpose was to find possible beneficial effects of plasmids that might offset their disadvantages. We found that, in all cases tested, plasmids conferred an advantage to growth at temperatures close to the top of the range for this fungus. Also, the plasmids improved fertility, as measured by perithecial production. Negative results were obtained for heavy metal resistance and ascospore germination. The results generate the hypothesis that plasmids may have adaptive significance to their hosts. PMID- 10686140 TI - Search for a novel killer toxin in yeast Pichia pastoris. AB - Certain yeast strains secrete a protein toxin, which inhibits the growth of sensitive pathogens and yeasts. Studies have shown that production of the toxin is dependent on presence of linear, double-stranded DNA plasmids in the killer yeasts. In the yeast Pichia pastoris, two linear double-stranded DNA plasmids have been identified. In the present study, the search for toxin-producing capability in P. pastoris has been conducted. No killer activity could be detected when 14 different indicator strains were tested. PMID- 10686141 TI - Single-step purification of recombinant human growth hormone (hGH) directly from bacterial osmotic shock fluids, for the purpose of (125)I-hGH preparation. AB - A good quality tracer, to be used in the radioimmunoassay of human growth hormone, was prepared by applying the chloramine-T iodination technique to the recombinant product obtained after a single-step high-performance size-exclusion chromatography purification of a bacterial osmotic shock fluid. The labeling reaction presented a yield of about 65% and the purified tracer exhibited an antibody binding of approximately 50% (NIDDK reference antiserum diluted 1:600,000). These values are very similar to those obtained by radioiodinating highly purified clinical-grade recombinant human growth hormone obtained from the same periplasmic extract after the regular six-step purification process. Both tracers provided the same accuracy, when evaluated with the use of commercial quality control samples in a classical radioimmunoassay methodology, their stability being practically identical: about 18% decrease in antibody binding after 2 months of storage at -20 degrees C. The novel approach permits the utilization of transformed Escherichia coli strains as a source of freshly prepared, radioiodination-grade recombinant proteins, capable of providing better reproducibility and reagent continuity. PMID- 10686142 TI - Production of fluorescent single-chain antibody fragments in Escherichia coli. AB - We describe a novel vector-host system suitable for the efficient preparation of fluorescent single-chain antibody Fv fragments (scFv) in Escherichia coli. The previously described pscFv1F4 vector used for the bacterial expression of functional scFv to the E6 protein of human papillomavirus type 16 was modified by appending to its C-terminus the green fluorescent protein (GFP). The expression of the scFv1F4-GFP fusion proteins was monitored by analyzing of the typical GFP fluorescence of the transformed cells under UV illumination. The brightest signal was obtained when scFv1F4 was linked to the cycle 3 GFP variant (GFPuv) and expressed in the cytoplasm of AD494(DE3) bacteria under control of the arabinose promoter. Although the scFv1F4 expressed under these conditions did not contain disulfide bridges, about 1% of the molecules were able to bind antigen. Fluorescence analysis of antigen-coated agarose beads incubated with the cytoplasmic scFv-GFP complexes showed that a similar proportion of fusions retained both E6-binding and green-light-emitting activities. The scFv1F4-GFPuv molecules were purified by affinity chromatography and successfully used to detect viral E6 protein in transfected COS cells by fluorescence microscopy. When an anti-beta-galactosidase scFv, which had previously been adapted to cytoplasmic expression at high levels, was used in this system, it was possible to produce large amounts of functional fluorescent antibody fragments. This indicates that these labeled scFvs may have many applications in fluorescence-based single-step immunoassays. PMID- 10686143 TI - Purification and characterization of macrodontain I, a cysteine peptidase from unripe fruits of Pseudananas macrodontes (Morr.) harms (Bromeliaceae). AB - A new papain-like cysteine peptidase isolated from fruits of Pseudananas macrodontes (Morr.) Harms, a species closely related to pineapple (Ananas comosus L.), has been purified and characterized. The enzyme, named macrodontain I, is the main proteolytic component present in fruit extracts and was purified by acetone fractionation followed by anion-exchange chromatography. Separation was improved by selecting both an adequate pH value and a narrow saline gradient. Optimum pH range (more than 90% of maximum activity with casein) was achieved at pH 6.1-8.5. Homogeneity of the enzyme was confirmed by bidimensional electrophoresis and mass spectroscopy (MS). Molecular mass of the enzyme was 23,459 (MS) and its isoelectric point was 6.1. The alanine, glutamine, and tyrosine derivatives were strongly preferred when the enzyme was assayed on N alpha-CBZ-l-amino acid p-nitrophenyl esters. The N-terminal sequence of macrodontain (by comparison with the N-terminus of 30 plant proteases with more than 50% homology) showed a great deal of sequence similarity to the other pineapple-stem-derived cysteine endopeptidases, being 85.7, 85. 2, and 77.8% identical to comosain, stem bromelain, and ananain, respectively. It seems clear that the Bromeliaceae endopeptidases are more closely related to each other than to other members of the papain family, suggesting relatively recent divergence. PMID- 10686144 TI - Production, purification, and properties of an extracellular laccase from Rigidoporus lignosus. AB - Laccase from Rigidoporus lignosus, a white-rot basidiomycete, has been isolated from culture filtrates. The enzyme was purified to homogeneity and some of its structural and kinetic parameters have been determined. The effects of pH, temperature, and organic solvents on the activity and stability of the enzyme, under different conditions, were also assayed. The results we have obtained, including the rather broad substrate specificity of enzyme, combined with their relatively easy production and purification, suggest that laccase may be efficiently employed in a variety of biotechnology applications. PMID- 10686145 TI - Replication-associated activities of purified human papillomavirus type 11 E1 helicase. AB - Replication of human papillomavirus type11 (HPV11) requires both the E1 and the E2 proteins. E1 is structurally and functionally similar to SV40 large T-antigen and is a DNA helicase/NTPase that binds to the origin of replication and initiates viral DNA replication. The biochemical characterization of HPV E1 is incompletely documented in the literature in part because of difficulties in expressing and purifying the protein. Herein, we report a method for the overexpression of full-length, untagged E1 (73.5 kDa) in baculovirus-infected Trichoplusia ni insect cells and the purification to homogeneity using a two-step procedure. The purified protein is a nonspecific NTPase that hydrolyzes ATP, dATP, UTP, or GTP equally well. Point mutations were made in the putative NTPase domain to verify that the activities observed were encoded by E1. Purified mutant D523N had negligible ATPase and helicase activities but retained DNA-binding activity. Sedimentation equilibrium ultracentrifugation and glycerol gradient centrifugation demonstrated that the wild-type protein is primarily a hexamer in its purified form. Secondary structure determination by circular dichroism revealed a large percentage of alpha-helical structure consistent with secondary structure predictions. These data define a fundamental set of biochemical and kinetic parameters for HPV E1 which are a critical prerequisite to future mechanistic studies of the enzyme. PMID- 10686146 TI - Isolation and characterization of bovine thymus multicatalytic proteinase complex. AB - The multicatalytic proteinase complex (MPC or proteasome) from bovine thymus was isolated and purified to homogeneity applying a protocol utilizing ion exchange and gel permeation chromatography as major purification tools. The purified complex shows molecular properties that are common for proteasomal molecules (high molecular mass, multisubunit organization, and multiple proteolytic activities) even though a peculiar subunit composition and the presence of specific regulatory mechanisms affecting the assembled proteolytic activities suggest a specialized function for this complex. Thymus proteasome is characterized by the presence of LMP2, LMP7, and LMP10 (MECL1) subunits, which replace the X, Y, and Z subunits. Since a similar complex was previously isolated in bovine spleen, it appears that the proteasomal population containing the LMP subunits is characteristic for organs involved in immune response. Both the thymus and spleen proteasomes are characterized by a marked efficiency in cleaving peptide bonds after branched-chain and aromatic amino acids, indicating that this proteasomal population is most likely involved in intracellular processing of class I antigenic peptides and is an example of an "in vivo" functioning immunoproteasome. However, in spite of several similarities, the complexes isolated from the two lymphoid organs do not show superimposable functional properties, which suggests the presence of organ-specific regulatory mechanisms affecting each of the proteolytic components assembled in the complex. PMID- 10686147 TI - Membrane-bound human 3beta-hydroxysteroid dehydrogenase: overexpression with His tag using a baculovirus system and single-step purification. AB - The membrane-bound human 3beta-hydroxysteroid dehydrogenase type 1 (3beta-HSD1) was overexpressed with His(6)-tag, using a baculovirus expression system, and then purified by nickel-chelated affinity chromatography. Overexpression of 3beta HSD1 was confirmed by enzyme assay and Western blot analysis. The protein was purified to more than 95% homogeneity by a single-step Ni(2+)-chelated affinity chromatography after solubilization of the membrane-bound protein with the detergent C(12)E(8). High yield was repeatedly obtained, with 3-4 mg of homogeneous and active 3beta-HSD1 from 1 x 10(9) of infected Sf9 cells. The kinetic study showed a K(m) of 1.7 microM and a V(max) of 50 nmol/min/mg of purified protein using dehydroepiandrosterone as the substrate. The above preparation will facilitate the structure-function study of this important enzyme. PMID- 10686148 TI - Extracellular expression, purification, and characterization of a winter flounder antifreeze polypeptide from Escherichia coli. AB - HPLC6 is the major component of liver-type antifreeze polypeptides (AFPs) from the winter flounder, Pleuronectes americanus. To facilitate mutagenesis studies of this protein, a gene encoding the 37-amino acid mature polypeptide was chemically synthesized and cloned into the Tac cassette immediately after the bacterial ompA leader sequence for direct excretion of the AFP into the culture medium. Escherichia coli transformant with the construct placIQpar8AF was cultured in M9 medium. The recombinant AFP (rAFP) was detected by a competitive enzyme-linked immunosorbent assay (ELISA). After IPTG induction, a biologically active rAFP was expressed. The majority of the rAFP was excreted into the culture medium with only trace amounts trapped in the periplasmic space and cytoplasm. After 18 h of induction, the accumulated rAFP in the culture medium amounted to about 16 mg/L. The excreted AFP was purified from the culture medium by a single step reverse-phase HPLC. Mass spectrometric and amino acid composition analyses confirmed the identity of the purified product. The rAFP, which lacked amidation at the C-terminal, was about 70% active when compared to the amidated wild-type protein, thus confirming the importance of C-terminal cap structure in protein stability and function. PMID- 10686149 TI - Optimization of inclusion body solubilization and renaturation of recombinant human growth hormone from Escherichia coli. AB - Recombinant human growth hormone (r-hGH) was expressed in Escherichia coli as inclusion bodies. In 10 h of fed-batch fermentation, 1.6 g/L of r-hGH was produced at a cell concentration of 25 g dry cell weight/L. Inclusion bodies from the cells were isolated and purified to homogeneity. Various buffers with and without reducing agents were used to solubilize r-hGH from the inclusion bodies and the extent of solubility was compared with that of 8 M urea as well as 6 M Gdn-HCl. Hydrophobic interactions as well as ionic interactions were found to be the dominant forces responsible for the formation of r-hGH inclusion bodies during its high-level expression in E. coli. Complete solubilization of r-hGH inclusion bodies was observed in 100 mM Tris buffer at pH 12.5 containing 2 M urea. Solubilization of r-hGH inclusion bodies in the presence of low concentrations of urea helped in retaining the existing native-like secondary structures of r-hGH, thus improving the yield of bioactive protein during refolding. Solubilized r-hGH in Tris buffer containing 2 M urea was found to be less susceptible to aggregation during buffer exchange and thus was refolded by simple dilution. The r-hGH was purified by use of DEAE-Sepharose ion-exchange chromatography and the pure monomeric r-hGH was finally obtained by using size exclusion chromatography. The overall yield of the purified monomeric r-hGH was approximately 50% of the initial inclusion body proteins and was found to be biologically active in promoting growth of rat Nb2 lymphoma cell lines. PMID- 10686150 TI - Expression of biologically active recombinant pokeweed antiviral protein in methylotrophic yeast Pichia pastoris. AB - Pokeweed antiviral protein (PAP)-I from the spring leaves of Phytolacca americana is a naturally occurring RNA-depurinating enzyme with broad-spectrum antiviral activity. Interest in PAP is growing due to its use as a potential anti-HIV agent. However, the clinical use of native PAP is limited due to inherent difficulties in obtaining sufficient quantities of homogeneously pure active PAP without batch-to-batch variation from its natural resource. Here, we report the expression of mature PAP (residues 23 to 284) with a C-terminal hexahistidine tag in the methylotrophic yeast Pichia pastoris, as a secreted soluble protein. The final yield of the secreted PAP is greater than 10 mg/L culture in shaker flasks. The secreted recombinant protein is not toxic to the yeast cells and has an apparent molecular mass of 33-kDa on SDS-PAGE gels. The in vitro enzymatic activity and cellular anti-HIV activity of recombinant PAP were of the same magnitude as those of the native PAP purified from P. americana. To our knowledge, this is the first large-scale expression and purification of soluble and biologically active recombinant mature PAP from yeast. PMID- 10686151 TI - Vectors allowing amplified expression of the Saccharomyces cerevisiae Gal3p Gal80p-Gal4p transcription switch: applications to galactose-regulated high-level production of proteins. AB - The Gal4, Gal80, and Gal3 proteins of Saccharomyces cerevisiae constitute a galactose-responsive regulatory switch for GAL gene promoters. The low cellular levels of these proteins have hampered mechanistic studies and limit the utility of the GAL gene promoters for high-yield production of endogenous and exogenous proteins. We have constructed two new vectors, pMEGA2 and pMEGA2-DeltaURA3, that increase the level of the Gal4p-Gal80p-Gal3p switch proteins under conditions that preserve the Gal3p-Gal80p-Gal4p stoichiometries required for normal switch function. Cells carrying pMEGA2 show 15- to 20-fold more Gal4p and 30- to 40-fold more Gal3p and Gal80p than cells lacking pMEGA2. These high levels of Gal4p, Gal80p, and Gal3p do not perturb the integrity of galactose-inducible regulation. Cells that carry pMEGA2 exhibit normal galactose-induction kinetics for the chromosomal MEL1 gene expression and normal, albeit slower, log-phase growth. Insertion of the MEL1 gene into pMEGA2 provides a 24- to 30-fold increase in the Mel1 protein. Cells carrying a 2-microm-based URA3-selectable plasmid containing a GAL1pro:lacZ reporter gene and a second plasmid, pMEGA2-DeltaURA3, produce 12 fold more beta-galactosidase than cells carrying only the GAL1pro:lacZ reporter plasmid. The performance of the MEGA plasmids in providing amplified production of the Gal3, Gal80, and Gal4 proteins should prove useful in investigations of the mechanistic aspects of these transcription switch proteins and in work aimed at achieving high-level, galactose-regulatable production of proteins in yeast. PMID- 10686152 TI - Expression, purification, and characterization of human recombinant thrombopoietin in Chinese hamster ovary cells. AB - Thrombopoietin (TPO) is a primary regulator of megakaryocytopoiesis, a process through which megakaryocytes proliferate and mature into platelets. Recombinant human TPO (rhTPO) was expressed in Chinese hamster ovary (CHO) cells and purified from the culture medium. The cDNA encoding full-length TPO, including the native signal peptide sequence, was amplified by PCR from a human fetal liver cDNA library. The product was cloned into a mammalian expression vector under the control of the SV40 early promoter and enhancer. Secreted rhTPO was purified in three conventional chromatography steps. It migrates on SDS-PAGE as a broad band, characteristic of a heavily glycosylated protein, with an average molecular mass of 85 kDa. rhTPO expressed in CHO cells is biologically active in vitro as demonstrated by its ability to stimulate the proliferation of a megakaryocytic cell line and to trigger the JAK/STAT signal transduction pathway. rhTPO also shows activity in vivo as judged by the elevation of platelet count in treated mice. PMID- 10686153 TI - Expression of functional recombinant antibody molecules in insect cell expression systems. AB - Recombinant single-chain variable-fragment molecules (scFv) were constructed from a cell line expressing a monoclonal antibody against African cassava mosaic virus (ACMV) and expressed in Escherichia coli. DNA sequences that encoded the scFv were manipulated to allow scFv expression in insect cell lines. A recombinant baculovirus containing the scFv cDNA was constructed and large amounts of scFv were produced in each of three insect cell lines infected with the baculovirus. However, the scFv were not secreted into the medium by any of the cell lines despite the scFv having been linked to a honeybee melittin leader sequence. The same scFv cDNA construct was introduced into Drosophila DS2 cells and a stable recombinant cell line was obtained that produced scFv that was secreted into the medium. Culture medium containing the scFv was used directly in enzyme-linked immunosorbent assay (ELISA) tests to detect ACMV in plant tissues. Another construct that encoded the Ckappa domain of human IgG was fused to the C-terminus of the scFv that was produced and expressed in Drosophila cells. This scFv derivative also accumulated in the medium and was more active in ELISA than scFv lacking the Ckappa domain. PMID- 10686154 TI - Purification of myelin basic protein from bovine brain. AB - Myelin basic protein (MBP) is a commonly used substrate for in vitro determination of numerous protein kinase activities. Herein we describe a rapid method for isolating relatively large amounts of MBP from bovine brain with a purity greater than that currently available from commercial sources. Lipids were first extracted from the CNS tissue by homogenization in sec-butanol. Washes under neutral and mildly basic conditions were employed to remove neutral and acidic proteins from the defatted residue. MBP was subsequently extracted under acidic conditions and further purified by chromatography on CM Sephadex C-25. Potential contaminating enzyme activities were destroyed by heart treatment. This method typically yields a recovery of 1.0-1.5 mg MBP per gram of starting material with a purity of greater than 95%. The MBP prepared in this manner was suitable for determination of kinase activities by both solution and the "in gel" kinase assay systems. PMID- 10686155 TI - Reconstitution of bacterial expressed human CD94: the importance of the stem region for dimer formation. AB - Human CD94 is a subunit of the disulfide-linked, heterodimeric natural killer (NK) cell surface receptor CD94/NKG2. This receptor, a member of the C-type lectin superfamily, participates in regulating NK cell directed lysis through interaction with the major histocompatibility antigen HLA-E. Two forms of CD94 were expressed using a bacterial expression system and refolded in vitro. One form, residues 34-179, designated S34, corresponds to the entire extracellular region of the receptor, including a 23-residue stem region, and the other, residues 51-179, designated E51, corresponds only to the putative carbohydrate recognition domain of the receptor. The refolded full-length S34 protein existed as a noncovalent dimer initially but formed an interchain disulfide bond upon storage for several months. In contrast, the stemless construct, E51, existed largely as a monomeric form. The stem region of S34, residues 34-56, is sensitive to proteolysis and its absence results in dissociation of the dimer. This suggests that the residues in the stem region of CD94 help to stabilize the dimeric conformation. PMID- 10686156 TI - Staphylococcal protein A as a fusion partner directs secretion of the e1alpha and e1beta subunits of pea mitochondrial pyruvate dehydrogenase by Bacillus subtilis. AB - Staphylococcal protein A (SPA)-based vectors were constructed to direct secretion of the E1alpha and E1beta subunits of Pisum sativum mitochondrial pyruvate dehydrogenase from Bacillus subtilis. These proteins were not exported when the signal peptide from levansucrase (SacBSP) was fused to their N-termini. Both SacBSP-E1alpha and SacBSP-E1beta fusion proteins were insoluble in the cytoplasm. However, when the SPA open-reading frame was inserted between SacBSP and E1alpha or E1beta, corresponding fusion proteins were secreted from the cells. The first (E) IgG-binding domain of SPA was sufficient to direct low level secretion of both fusion proteins (SacBSP-E-E1alpha and SacBSP-E-E1beta). Adding the second (D) IgG-binding domain improved extracellular protein yields 3- to 4-fold over E alone, but was not as efficient as secretion of the full-length (EDABC) SPA fusion proteins. All constructs were based on the pUB110-derived multicopy plasmid pWB705. Separate B. subtilis strains transformed with SacBSP-E-E1alpha His(6) or SacBSP-E1beta were cocultivated in the presence of Ni-NTA agarose. The native pyruvate dehydrogenase alpha2beta2 structure was bound to the affinity matrix, demonstrating assembly after secretion. The use of SPA as a fusion partner during expression of heterologous proteins by B. subtilis provides the basis of a versatile system that can be used to study both secretion and protein:protein interactions. PMID- 10686157 TI - Trophic Cascades in Terrestrial Systems: A Review of the Effects of Carnivore Removals on Plants. AB - We present a quantitative synthesis of trophic cascades in terrestrial systems using data from 41 studies, reporting 60 independent tests. The studies covered a wide range of taxa in various terrestrial systems with varying degrees of species diversity. We quantified the average magnitude of direct effects of carnivores on herbivore prey and indirect effects of carnivores on plants. We examined how the effect magnitudes varied with type of carnivores in the study system, food web diversity, and experimental protocol. A metaanalysis of the data revealed that trophic cascades were common among the studies. Exceptions to this general trend did arise. In some cases, trophic cascades were expected not to occur, and they did not. In other cases, the direct effects of carnivores on herbivores were stronger than the indirect effects of carnivores on plants, indicating that top down effects attenuated. Top-down effects usually attenuated whenever plants contained antiherbivore defenses or when herbivore species diversity was high. Conclusions about the strength of top-down effects of carnivores varied with the type of carnivore and with the plant-response variable measured. Vertebrate carnivores generally had stronger effects than invertebrate carnivores. Carnivores, in general, had stronger effects when the response was measured as plant damage rather than as plant biomass or plant reproductive output. We caution, therefore, that conclusions about the strength of top-down effects could be an artifact of the plant-response variable measured. We also found that mesocosm experiments generally had weaker effect magnitudes than open-plot field experiments or observational experiments. Trophic cascades in terrestrial systems, although not a universal phenomenon, are a consistent response throughout the published studies reviewed here. Our analysis thus suggests that they occur more frequently in terrestrial systems than currently believed. Moreover, the mechanisms and strengths of top-down effects of carnivores are equivalent to those found in other types of systems (e.g., aquatic environments). PMID- 10686158 TI - The Effects of a Bottleneck on Inbreeding Depression and the Genetic Load. AB - We study the effects of a population bottleneck on the inbreeding depression and genetic load caused by deleterious mutations in an outcrossing population. The calculations assume that loci have multiplicative fitness effects and that linkage disequilibrium is negligible. Inbreeding depression decreases immediately after a sudden reduction of population size, but the drop is at most only several percentage points, even for severe bottlenecks. Highly recessive mutations experience a purging process that causes inbreeding depression to decline for a number of additional generations. On the basis of available parameter estimates, the absolute fall in inbreeding depression may often be only a few percentage points for bottlenecks of 10 or more individuals. With a very high lethal mutation rate and a very slow population growth, however, the decline may be on the order of 25%. We examine when purging might favor a switch from outbreeding to selfing and find it occurs only under very limited conditions unless population growth is very slow. In contrast to inbreeding depression, a bottleneck causes an immediate increase in the genetic load. Purging causes the load to decline and then overshoot its equilibrium value. The changes are typically modest: the absolute increase in the total genetic load will be at most a few percentage points for bottlenecks of size 10 or more unless the lethal mutation rate is very high and the population growth rate very slow. PMID- 10686159 TI - Seed Germination in Desert Annuals: An Empirical Test of Adaptive Bet Hedging. AB - Temporal variability in survivorship and reproduction is predicted to affect the evolution of life-history characters. Desert annual plants experience temporal variation in reproductive success that is largely caused by precipitation variability. We studied several populations of the desert annual Plantago insularis along a precipitation gradient. Whereas models of bet hedging in unpredictable environments generally predict one optimal germination fraction for a population, empirical studies have shown that environmental conditions during germination can cause a range of germination fractions to be expressed. In a 4-yr field study, we found that populations in historically more xeric environments had lower mean germination fractions, as is predicted by bet-hedging models. However, populations exhibited significant variation in germination among years. Two experimental studies measuring germination under several environment conditions were conducted to elucidate the source of this in situ variation. Germination fractions exhibited phenotypic plasticity in response to water availability and date within the season. Populations differed in their norms of reaction such that seeds from more xeric populations germinated under less restrictive conditions. A pattern of delayed germination consistent with among year bet-hedging predictions arose in the field through the interaction of seed germinability and the distribution of environmental conditions during germination. PMID- 10686160 TI - Resource Allocation and the Evolution of Self-Fertilization in Plants. AB - This article develops a simple evolutionarily stable strategy (ESS) model of resource allocation in partially selfing plants, which incorporates reproductive and sex allocation into a single framework. The analysis shows that, if female fitness gain increases linearly with resource investment, total reproductive allocation is not affected by sex allocation, defined as the fraction of reproductive resources allocated to male function. All else being equal, the ESS total reproductive allocation increases with increasing selfing rate if the fitness of selfed progeny is more than half that of outcrossed progeny, while the ESS sex allocation is always a decreasing function of the selfing rate. Self fertilization is much more common in annual than in perennial plants, and this association has been commonly interpreted in terms of an effect of life history on mating system. The model in this article shows that self-fertilization can itself cause the evolution of the annual habit. Incorporating the effects of pollen discounting may not have any influence on total reproductive allocation if female fitness gain is a linear function of resource investment, although the evolutionarily stable sex allocation is altered. Evolution of the selfing rate is found to be independent of reproductive and sex allocation under the mass-action assumption that self- and outcross pollen are deposited simultaneously on receptive stigmas and compete for access to ovules. PMID- 10686161 TI - Effects of Enrichment on Three-Level Food Chains with Omnivory. AB - Although omnivory (the consumption of resources from more than one trophic level) is widespread, this fundamental limitation to the applicability of food chain theory to real communities has received only limited treatment. We investigated effects of enrichment (increasing carrying capacity, K, of the resource) on a system consisting of a resource (R), an intermediate consumer (N), and an omnivore (P) using a general mathematical model and tested the relevance of some of its predictions to a laboratory system of mixed bacteria (=R) and the ciliates Tetrahymena (=N) and Blepharisma (=P). The model produced six major predictions. First, N may facilitate or inhibit P. Enrichment may revert the net effect of N on P from facilitation to inhibition. Second, along a gradient of K, up to four regions of invasibility and stable coexistence of N and P may exist. At the lowest K, only R is present. At somewhat higher K, N can coexist with R. At intermediate K, either N and P coexist, or either consumer excludes the other depending on initial conditions. At the highest K, N may be excluded through apparent competition and only R and P can coexist. The pattern of persistence of Tetrahymena and Blepharisma along an enrichment gradient conformed fairly well to the scenario allowing coexistence at intermediate K. Third, for stable equilibria of the omnivory system, R always increases and N always decreases with K. The abundances of bacteria and Tetrahymena were suggestive of such a pattern but did not allow a strict test because coexistence occurred at only one level of enrichment. Fourth, an omnivore can invade an R-N system at a lower K than an otherwise identical specialist predator of N. Fifth, an omnivore can always invade a food chain with such a specialist predator. Sixth, over ranges of K where both omnivory systems and otherwise identical three-level food chains are feasible, N is always less abundant in the omnivory system, whereas the relative abundances of R and P in omnivory systems compared to food chains may change with K. It is thus possible that total community biomass at a given K is lower in an omnivory system than in a food chain. Both the model and the experimental results caution that patterns of trophic-level abundances in response to enrichment predicted by food chain theory are not to be expected in systems with significant omnivory. PMID- 10686162 TI - Dwarfs and Giants: Cannibalism and Competition in Size-Structured Populations. AB - Cannibals and their victims often share common resources and thus potentially compete. Smaller individuals are often competitively superior to larger ones because of size-dependent scaling of foraging and metabolic rates, while larger ones may use cannibalism to counter this competition. We study the interplay between cannibalism and competition using a size-structured population model in which all individuals consume a shared resource but in which larger ones may cannibalize smaller conspecifics. In this model, intercohort competition causes single-cohort cycles when cannibalism is absent. Moderate levels of cannibalism reduce intercohort competition, enabling coexistence of many cohorts. More voracious cannibalism, in combination with competition, produces large-amplitude cycles and a bimodal population size distribution with many small and few giant individuals. These coexisting ''dwarfs'' and ''giants'' have very different life histories, resulting from a reversal in importance of cannibalism and competition. The population structure at time of birth determines whether individuals suffer severe cannibalism, with the few survivors reaching giant sizes, or whether they suffer intense intracohort competition, with all individuals remaining small. These model results agree remarkably well with empirical data on perch population dynamics. We argue that the induction of cannibalistic giants in piscivorous fish is a population-dynamic emergent phenomenon that requires a combination of size-dependent cannibalism and competition. PMID- 10686163 TI - Trophic Interactions during Primary Succession: Herbivores Slow a Plant Reinvasion at Mount St. Helens. AB - Lupines (Lupinus lepidus var. lobbii), the earliest plant colonists of primary successional habitats at Mount St. Helens, were expected to strongly affect successional trajectories through facilitative effects. However, their effects remain localized because initially high rates of reinvasive spread were short lived, despite widespread habitat availability. We experimentally tested whether insect herbivores, by reducing plant growth and fecundity at the edge of the expanding lupine population, could curtail the rate of reinvasion and whether those herbivores had comparable impacts in the older, more successionally advanced core region. We found that removing insect herbivores increased both the areal growth of individual lupine plants and the production of new plants in the edge region, thereby accelerating the lupine's intrinsic rate of increase at the front of the lupine reinvasion. We found no such impacts of herbivory in the core region, where low plant quality or a complex of recently arrived natural enemies may hold herbivores in check. In the context of invasion theory, herbivore mediated decreases in lupine population growth rate in the edge region translate into decreased rates of lupine spread, which we quantify here using diffusion models. In the Mount St. Helens system, decreased rate of lupine reinvasion will result in reductions in rates of soil formation, nitrogen input, and entrapment of seeds and detritus that are likely to postpone or alter trajectories of primary succession. If the type of spatial subtleties in herbivore effects we found here are common, with herbivory focused on the edge of an expanding plant population and suppressed or ineffective in the larger, denser central region (where the plants might be more readily noticed and studied), then insect herbivores may have stronger impacts on the dynamics of primary succession and plant invasions than previously recognized. PMID- 10686164 TI - Population Dynamic and Genetic Consequences of Spatial Density-Dependent Dispersal in Patchy Populations. AB - Predictions about sex-specific, spatial density-dependent dispersal and their demographic and genetic consequences were tested in experimental populations of root voles (Microtus oeconomus). Each population consisted of two demes inhabiting equal-sized habitat patches imbedded in a barren matrix area. We used a neutral two-allele allozyme marker to monitor gene flow. Initially, the two demes were genetically distinct and had different densities so that the size of a high-density deme (genotype bb) was four times larger than that of a low-density deme (genotype aa). The sex-specific dispersal pattern was in accordance with our prediction. Male dispersal was unconditional on deme-specific densities, and the majority of the first-generation males became dispersed from both demes, whereas female dispersal was strongly density dependent, so that dispersal took place exclusively from the high-density to the low-density deme. The demographic implication of this dispersal pattern was that the initial density difference between the demes was quickly canceled out. We built a mathematical model that predicted that the initially rare allele (a) would increase in frequency given the dispersal pattern, and this was supported by our experimental data. This result relies mostly on the density-independent male-dispersal strategy, which presumably stems from inbreeding avoidance. Our study highlights the importance of incorporating sex-specific dispersal strategies in population genetic models. Sex-biased dispersal may act as a deterministic force counteracting the tendency for stochastic loss of alleles in small and fragmented populations. PMID- 10686165 TI - Grazers and Diggers: Exploitation Competition and Coexistence among Foragers with Different Feeding Strategies on a Single Resource. AB - A mathematical model is presented that describes a system where two consumer species compete exploitatively for a single renewable resource. The resource is distributed in a patchy but homogeneous environment; that is, all patches are intrinsically identical. The two consumer species are referred to as diggers and grazers, where diggers deplete the resource within a patch to lower densities than grazers. We show that the two distinct feeding strategies can produce a heterogeneous resource distribution that enables their coexistence. Coexistence requires that grazers must either move faster than diggers between patches or convert the resources to population growth much more efficiently than diggers. The model shows that the functional form of resource renewal within a patch is also important for coexistence. These results contrast with theory that considers exploitation competition for a single resource when the resource is assumed to be well mixed throughout the system. PMID- 10686166 TI - Energy, Density, and Constraints to Species Richness: Ant Assemblages along a Productivity Gradient. AB - Species richness describes the number of species of a given taxon in a given time and space. The energy limitation hypothesis links the species richness of consumer taxa to net primary productivity (NPP) through two relationships: NPP limits a taxon's density, and taxon density limits species richness. We study both relationships with a survey of 15 ground ant assemblages, along a productivity gradient from deserts to rain forests. Ant density (colonies m-2) was a positive, decelerating function of net aboveground productivity (NAP). A stepwise regression suggests that the efficiency with which NAP is converted to ant colonies increases with maximum summer temperature and decreases with precipitation. Ant species richness was a positive decelerating function of density at three spatial scales. This supports the energy limitation hypothesis' assumption that average population densities are higher in environments that are more productive. These two nonlinear functions (NAP-density and density-species richness) combine to create, at a variety of scales, positive, decelerating, productivity-diversity curves for a common, ecologically dominant taxon across the terrestrial productivity gradient. However, variance in the density and diversity explained by NAP decreases with scale, suggesting that energy limitation of diversity predominates at small spatial scales (<1 ha). PMID- 10686167 TI - Nonlinear Dynamics and the Evolution of Semelparous and Iteroparous Reproductive Strategies. PMID- 10686168 TI - Unravelling the pathophysiology of calcium channel mutations causing neurological disorders. PMID- 10686169 TI - Tourette syndrome, associated conditions and the complexities of treatment. AB - Tourette syndrome (TS) is characterized by multiple motor tics plus one or more vocal (phonic) tics, which characteristically wax and wane. It can no longer be considered the rare and bizarre syndrome that it was once thought to be. The concepts surrounding TS, and our understanding of it, are also becoming increasingly complex and, in some individuals, TS is now recognized to be associated with a wide variety of associated behaviours and psychopathologies. It is suggested that TS is heterogeneous from a variety of standpoints including clinical presentation and psychopathology, and thus neuropharmacological responses and possibly even aetiological and genetic mechanisms. In this paper, mention is made of recent findings in epidemiology and genetics, highlighting the complexities of the disorder; these have been chosen because findings in both areas have clinical and management implications. The literature on the clinical manifestations, associated behaviours, psychopathology (and/or comorbid conditions) and management, in particular, is reviewed in detail. PMID- 10686170 TI - Abnormal transmitter release at neuromuscular junctions of mice carrying the tottering alpha(1A) Ca(2+) channel mutation. AB - Neurotransmitter release at many synapses is regulated by P/Q-type Ca(2+) channels containing the alpha(1A) pore-forming subunit. Mutations in alpha(1A) cause cerebral disorders including familial hemiplegic migraine (FHM) and ataxia in humans. Tottering (tg) alpha(1A) mutant mice display ataxia and epilepsy. It is not known whether alpha(1A) mutations induce impairment of synaptic function, which could underlie the symptoms of these cerebral disorders. To assess whether alpha(1A) mutations influence neurotransmitter release, we studied P-type Ca(2+) channel-mediated acetylcholine (ACh) release at tg neuromuscular junctions (NMJs) with micro-electrode measurements of synaptic potentials. We found a Ca(2+)-, Mg(2+)- and K(+)-dependent increase of spontaneous ACh release at both homo- and heterozygote tg NMJs. Furthermore, there was increased run-down of high-rate evoked release at homozygous tg NMJs. In isotonic contraction experiments this led to block of synaptic transmission at lower concentrations of the ACh antagonist tubocurarine than were needed in wild-type muscles. Our results suggest that in tg motor nerve terminals there is increased influx of Ca(2+) under resting conditions. This study shows that functional consequences of alpha(1A) mutations causing cerebral disorders can be characterized at the NMJ. PMID- 10686171 TI - Accelerated forgetting in patients with epilepsy: evidence for an impairment in memory consolidation. AB - Patients with epilepsy frequently complain of memory difficulties yet perform normally on standard neuropsychological tests of memory. It has been suggested that this may be due to an impairment of very long-term memory consolidation processes, beyond those normally assessed in the neuropsychological clinic. We carried out a prospective study of verbal memory over a long-term retention interval of 8 weeks in patients with epilepsy and in controls. Results were compared with performance on conventional tests of memory. Despite normal learning and retention over 30 min, patients with epileptic foci in the left temporal lobe performed disproportionately poorly on the long-term test compared with both patients with epileptic foci in the right temporal lobe and controls. Our findings provide evidence for an extended period of memory consolidation and point to the critical region for this process, at least for verbal material, in the left temporal lobe. The implications of our findings for clinical assessment and therapeutic management of patients with epilepsy are discussed. PMID- 10686172 TI - Atypical and typical presentations of Alzheimer's disease: a clinical, neuropsychological, neuroimaging and pathological study of 13 cases. AB - There has been increasing awareness that some slowly progressive focal cortical syndromes can be the presenting features of Alzheimer's disease, but pathological evidence has been sparse. This clinico-pathological series presents our experience with pathologically proven atypical as well as typical Alzheimer's disease presentations. We report and compare four patterns of presentation: a typical pattern with initial amnesic syndrome (n = 4 cases), progressive visual dysfunction (n = 1), progressive biparietal syndrome (n = 2) and progressive aphasia (n = 6). The aphasic presentations include both fluent and non-fluent aphasic syndromes. The neuropsychological profiles and neuroimaging clearly reflect the presenting clinical features, and show a close relationship to the distribution of pathology in these cases. Of note was the sparing of medial temporal structures (hippocampus and/or entorhinal cortex) in several aphasic cases and the severe occipito-parietal involvement in those with prominent visuospatial disorders at presentation. Our data demonstrate the wide spectrum of Alzheimer's disease presentations. The recognition of atypical presentations of Alzheimer's disease is important when attempting to make an early accurate pre morbid diagnosis of neurodegenerative disease. PMID- 10686173 TI - Developmental amnesia associated with early hypoxic-ischaemic injury. AB - We recently reported on three young patients with severe impairments of episodic memory resulting from brain injury sustained early in life. These findings have led us to hypothesize that such impairments might be a previously unrecognized consequence of perinatal hypoxic-ischaemic injury. Neuropsychological and quantitative magnetic resonance investigations were carried out on five young patients, all of whom had suffered hypoxic-ischaemic episodes at or shortly after birth. All five patients showed severe impairments of episodic memory (memory for events), with relative preservation of semantic memory (memory for facts). However, none had any of the major neurological deficits that are typically associated with hypoxic-ischaemic injury, and all attended mainstream schools. Quantitative magnetic resonance investigations revealed severe bilateral hippocampal atrophy in all cases. As a group, the patients also showed bilateral reductions in grey matter in the regions of the putamen and the ventral part of the thalamus. On the basis of their clinical histories and the pattern of magnetic resonance findings, we attribute the patients' pathology and associated memory impairments primarily to hypoxic-ischaemic episodes sustained very early in life. We suggest that the degree of hypoxia-ischaemia was sufficient to produce selective damage to particularly vulnerable regions of the brain, notably the hippocampi, but was not sufficient to result in the more severe neurological and cognitive deficits that can follow hypoxic-ischaemic injury. The impairments in episodic memory may be difficult to recognize, particularly in early childhood, but this developmental amnesia can have debilitating consequences, both at home and at school, and may preclude independent life in adulthood. PMID- 10686174 TI - Repertoire dynamics of autoreactive T cells in multiple sclerosis patients and healthy subjects: epitope spreading versus clonal persistence. AB - Autoantigen-specific T-lymphocytes are present in patients with autoimmune disease and in normal subjects. Little is currently known about the temporal variation (dynamics) of the immune repertoire of these autoreactive T cells. We analysed the long-term variation of the immune repertoire of T cells specific for myelin basic protein (MBP) in five untreated patients with multiple sclerosis and four normal control subjects over a mean observation period of 6 years. MBP specific CD4(+) T-cell lines were selected with purified human MBP, and their epitope specificity was mapped with overlapping synthetic peptides. Three distinct patterns of repertoire development were observed. (i) Two patients and three control subjects maintained a broad epitope response with fluctuations over time. (ii) Two patients initially showed a focused response that broadened over the course of 6 years; this finding could be described as intramolecular epitope spreading. (iii) In one patient and one control subject, a strikingly focused response, which was directed to a cluster of nested epitopes in the MBP region 83 102, persisted over time. T-cell receptor Vbeta sequence analysis allowed us to trace individual clones of MBP-specific T cells for up to 7 years in the peripheral circulation in four of the five patients and three of the four controls, suggesting that the long-term persistence of MBP-specific T-cell clones is a common feature of the T-cell repertoire not unique to multiple sclerosis. The persisting MBP-specific T-cell clones were not detectable in the blood of one of the patients by complementarity-determining region (CDR)-3 spectratyping, indicating that their frequency does not exceed 1 in 5000 T cells. The temporal characteristics of the MBP-specific T-cell repertoire described here are relevant to therapeutic strategies targeting autoantigen-specific T cells in multiple sclerosis and other autoimmune diseases. PMID- 10686176 TI - The neural correlates of 'deaf-hearing' in man: conscious sensory awareness enabled by attentional modulation. AB - Attentional modulation of normal sensory processing has a two-fold impact on human brain activity: activation of a network of localized brain regions is associated with paying attention, and activation of specific sensory regions is enhanced relative to passive stimulation. The mechanisms underlying attentional modulation of perception in patients with lesions of sensory cortices are less well understood. Here we report a unique patient suffering from extensive bilateral destruction of the auditory cortices (including the primary auditory fields) who demonstrated conscious perception of the onset and offset of sounds only when selectively attending to the auditory modality. This is the first description of such an attentively modulated 'deaf-hearing' phenomenon and its neural correlates, using H(2)(15)O-PET. Increases in cerebral blood flow associated with conscious awareness of sound that was achieved by listening attentively (compared with identical auditory stimulation presented when the patient was inattentive) were found bilaterally in the lateral (pre)frontal cortices, the spared middle temporal cortices and the cerebellar hemispheres. We conclude that conscious awareness of sounds may be achieved in the absence of the primary auditory cortex, and that selective, 'top-down' attention, associated with prefrontal systems, exerts a crucial modulatory effect on auditory perception within the remaining auditory system. PMID- 10686175 TI - Axonal loss results in spinal cord atrophy, electrophysiological abnormalities and neurological deficits following demyelination in a chronic inflammatory model of multiple sclerosis. AB - Recent pathological studies have re-emphasized that axonal injury is present in patients with multiple sclerosis, the most common demyelinating disease of the CNS in humans. However, the temporal profile of demyelination and axonal loss in multiple sclerosis patients and their independent contributions to clinical and electrophysiological abnormalities are not completely understood. In this study, we used the Theiler's murine encephalomyelitis virus model of progressive CNS inflammatory demyelination to demonstrate that demyelination in the spinal cord is followed by a loss of medium to large myelinated fibres. By measuring spinal cord areas, motor-evoked potentials, and motor coordination and balance, we determined that axonal loss following demyelination was associated with electrophysiological abnormalities and correlated strongly with reduced motor coordination and spinal cord atrophy. These findings demonstrate that axonal loss can follow primary, immune-mediated demyelination in the CNS and that the severity of axonal loss correlates almost perfectly with the degree of spinal cord atrophy and neurological deficits. PMID- 10686177 TI - Receptive amusia: evidence for cross-hemispheric neural networks underlying music processing strategies. AB - Perceptual musical functions were investigated in patients suffering from unilateral cerebrovascular cortical lesions. Using MIDI (Musical Instrument Digital Interface) technique, a standardized short test battery was established that covers local (analytical) as well as global perceptual mechanisms. These represent the principal cognitive strategies in melodic and temporal musical information processing (local, interval and rhythm; global, contour and metre). Of the participating brain-damaged patients, a total of 69% presented with post lesional impairments in music perception. Left-hemisphere-damaged patients showed significant deficits in the discrimination of local as well as global structures in both melodic and temporal information processing. Right-hemisphere-damaged patients also revealed an overall impairment of music perception, reaching significance in the temporal conditions. Detailed analysis outlined a hierarchical organization, with an initial right-hemisphere recognition of contour and metre followed by identification of interval and rhythm via left hemisphere subsystems. Patterns of dissociated and associated melodic and temporal deficits indicate autonomous, yet partially integrated neural subsystems underlying the processing of melodic and temporal stimuli. In conclusion, these data contradict a strong hemispheric specificity for music perception, but indicate cross-hemisphere, fragmented neural substrates underlying local and global musical information processing in the melodic and temporal dimensions. Due to the diverse profiles of neuropsychological deficits revealed in earlier investigations as well as in this study, individual aspects of musicality and musical behaviour very likely contribute to the definite formation of these widely distributed neural networks. PMID- 10686178 TI - Encoding of burning pain from capsaicin-treated human skin in two categories of unmyelinated nerve fibres. AB - Burning pain was induced in healthy human subjects by intracutaneous injections of capsaicin (20 microl, 0.1%) in the innervation territory of the cutaneous branch of the peroneal nerve and the pain responses were compared with the activation patterns of afferent C-fibres recorded by microneurography. Responsiveness of single units to mechanical or heat stimuli or to sympathetic reflex provocation tests was determined by transient slowing of conduction velocity following activation (marking technique). Capsaicin activated each of 12 mechano-responsive and 17 of 20 mechano-insensitive C-units. However, the duration of the responses to capsaicin was significantly longer in mechano insensitive C-units (median 170 s; quartiles 80-390) compared with mechano responsive C-units (8 s; 4-10). The activation times of mechano-insensitive C units closely matched the duration of capsaicin-induced pain responses, whereas activation of mechano-responsive C-units was too short to account for the duration of the burning pain. The latter generally were desensitized to mechanical stimulation at the injection site, whereas 8 of 17 of the originally mechano-insensitive C-units became responsive to mechanical probing at the injection site after capsaicin. Responses typically started several seconds after the onset of the mechanical stimulus in parallel with pain sensations. We did not observe sensitization to brushing or to punctate stimuli in uninjured parts of the innervation territory. Differential capsaicin sensitivity adds to the cumulating evidence for the existence of two categories of functionally different nociceptors in human skin, with a special role for mechano-insensitive fibres in sensitization and hyperalgesia. Possible structural differences between these two categories are discussed, including the role of tetrodotoxin-resistant sodium channels. PMID- 10686179 TI - Induction of plasticity in the human motor cortex by paired associative stimulation. AB - Current models of motor cortical plasticity, developed in studies on experimental animals, emphasize the importance of the conjoint activity of somatosensory afferents and intrinsic motor cortical circuits. The hypothesis that an enduring change in excitability in the cortical output circuitry can be induced in the human motor cortex by a paired-stimulation protocol was tested. Low-frequency median nerve stimulation was paired with transcranial magnetic stimulation (TMS) over the optimal cranial site for stimulating the abductor pollicis brevis muscle (APB). This protocol induced an increase in the amplitudes of the motor evoked potentials (MEPs) in the resting APB as well as a prolongation of the silent period measured in the precontracted APB following TMS; amplitudes of MEPs measured in voluntary contraction remained unchanged. Experiments testing the excitability of spinal motoneurons using F-wave studies and electrical stimulation of the brainstem suggested that the site of the plastic changes was within the motor cortex. The increases in resting amplitudes and silent period duration were conditionally dependent on the timing between the afferent and the magnetic stimulation in that they were present when events elicited by afferent and magnetic stimulation were synchronous at the level of the motor cortex. Plasticity induced by paired stimulation evolved rapidly (within 30 min), was persistent (minimum duration 30-60 min) yet reversible, and was topographically specific. This combination of features and the similarity to properties of induced enduring changes in synaptic efficacy, as elucidated in animal studies, leads us to propose that the induced plasticity may represent a signature of associative long-term potentiation of cortical synapses or closely related neuronal mechanisms in the human cortex. PMID- 10686180 TI - The impact of apolipoprotein E genotypes on age at onset of symptoms and phenotypic expression in Wilson's disease. AB - Wilson's disease is a disorder of biliary copper excretion that may result in severe neurological symptoms and advanced liver disease. The wide variation of phenotypic disease expression cannot be fully explained by the different mutations of the Wilson disease gene. In neurological disorders, such as Alzheimer's disease, temporal lobe epilepsy and cerebral trauma, the presence of the apolipoprotein E (ApoE) allele epsilon4 is associated with an increased vulnerability of the brain to the effects of the disease, whereas the presence of the ApoE genotype epsilon3/3 appears to provide moderate neuroprotection. We examined whether this hypothesis holds true for the development of neurological symptoms in patients with Wilson's disease. The ApoE genotype and the H1069Q mutation (the most common in Wilson's disease) status were determined by polymerase chain reaction-based mutation assays in 121 well-characterized, symptomatic index patients with Wilson's disease. An investigation profile was established in which the patients were grouped according to the clinical symptoms at presentation, the ApoE genotypes and the status of the H1069Q mutation. Fifty nine per cent of the 121 patients had the allele combination ApoE epsilon3/3 (21% ApoE epsilon3/4, 19% ApoE epsilon3/2, 1% ApoE epsilon4/2). The distribution of ApoE genotypes did not deviate from known distributions in healthy European subjects. Within the group of 40 H1069Q-homozygous patients, the onset of symptoms was significantly delayed in patients with the ApoE epsilon3/3 genotype (25 +/- 6 years at presentation) compared with patients with the ApoE epsilon3/4 genotype (20 +/- 3 years at presentation). In this study, the ApoE genotype was established as an important factor delaying the onset of neurological and hepatic symptoms, but not modifying phenotypic disease expression in a homogeneous group of patients with Wilson's disease (all H1069Q-homozygotes, similar genetic background). The presence of ApoE epsilon3/3 attenuates clinical manifestations in Wilson's disease by mechanisms which might involve the antioxidant and membrane-stabilizing properties of the ApoE 3 protein. PMID- 10686181 TI - Cytochrome oxidase immunohistochemistry: clues for genetic mechanisms. AB - Cytochrome c oxidase (COX) is encoded by three mitochondrial and nine nuclear genes. COX deficiency is genetically heterogeneous but current diagnostic methods cannot easily distinguish between mitochondrial and nuclear defects. We hypothesized that there may be differential expression of COX subunits depending on the underlying mutation. COX subunit expression was investigated in five patients with known mtDNA mutations. Severe and selective reduction of mtDNA encoded COX subunits I and II was consistently observed in all these patients and was restricted to COX-deficient fibres. Immunostaining of nuclear-encoded subunits COX IV and Va was normal, whilst subunit VIc, also nuclear-encoded, was decreased. Twelve of 36 additional patients with histochemically defined COX deficiency also had this pattern of staining, suggesting that they had mtDNA defects. Clinical features in this group were heterogeneous, including infantile encephalopathy, multisystem disease, cardiomyopathy and childhood-onset isolated myopathy. The remaining patients did not have the same pattern of immunostaining. Fourteen had reduced staining of all subunits, whilst 10 had normal staining of all subunits despite reduced enzyme activity. Patients with COX deficiency secondary to mtDNA mutations have a specific pattern of subunit loss, but the majority of children with COX deficiency do not have this pattern of subunit loss and are likely to have nuclear gene defects. PMID- 10686182 TI - Cytoarchitectonic and immunohistochemical characterization of a specific pain and temperature relay, the posterior portion of the ventral medial nucleus, in the human thalamus. AB - Previous studies in the macaque monkey have identified a thalamic nucleus, the posterior portion of the ventral medial nucleus (VMpo), as a dedicated lamina I spinothalamocortical relay for pain and temperature sensation. The dense plexus of calbindin-immunoreactive fibres that characterizes VMpo in primates enables its homologue to be identified in the human thalamus by immunohistochemical labelling for calbindin. We have now analysed in detail the cytoarchitectonic characteristics of VMpo and its relationship with immunoreactivity for calbindin, substance P and calcitonin gene-related peptide (CGRP) in the human thalamus. The area in the posterolateral thalamus in which dense calbindin-immunoreactive fibre terminations are present coincides nearly completely with a distinct region that contains small to medium-sized cells with round or oval shapes that are aggregated in clusters separated by cell sparse areas. This region, which we identify as VMpo, is located posteromedial to the ventral posterior lateral (VPL) and ventral posterior medial (VPM) nuclei, ventral to the anterior pulvinar and centre median nuclei, lateral to the limitans and parafascicular nuclei and dorsal to the medial geniculate nucleus. Calbindin-immunoreactive fibres enter VMpo from the spinal lemniscus and form large patches of dense terminal-like staining over clusters of VMpo neurons. A few of these clusters also display terminal-like substance P labelling. Small bursts of CGRP staining are intercalated between the calbindin-labelled clusters, but there is little or no overlap between these two markers. CGRP immunoreactivity is also present over small, non-clustered neurons in the calbindin-negative area that separates VMpo from the VPL and VPM nuclei, which we denote as the posterior nucleus (Po). These observations provide a concise description of VMpo in the human thalamus. Further, they suggest that the lamina I spinothalamic tract fibres (represented by calbindin and probably also substance P immunoreactivity) and vagal-solitary parabrachial afferents (represented by CGRP immunoreactivity) form closely related, but separate, termination fields that can be considered to represent different aspects of enteroceptive information regarding the physiological status of the tissues and organs of the body. The location of VMpo and the adjacent Po fits with clinical descriptions of the thalamic area from which pain, temperature and visceral sensations can be evoked by microstimulation, and where nociceptive and thermoreceptive neurons have been recorded in humans. It also corresponds to the area in which infarcts cause analgesia and thermoanaesthesia and can lead to the paradoxical development of central pain. PMID- 10686183 TI - Functional MRI evidence for a role of frontal and inferior temporal cortex in amodal components of priming. AB - Changes in human brain activity associated with repetition priming during word generation were characterized across a series of neuroimaging and behavioural studies. Repetition priming was consistently observed behaviourally as a decrease in response latency for repeated items, and was found for both visually and aurally cued word-generation tasks. Brain imaging using whole-brain functional MRI identified neural correlates of these effects. The principal effect of priming was to reduce neural activity within regions that were already being used to perform the word-generation tasks. Repeated word generation in response to visual cues was correlated with anatomically selective reductions in activity within the left frontal cortex along the inferior frontal gyrus and inferior temporal regions and, to a lesser degree, in specific earlier visual regions. These reductions were reversed when new items were presented, indicating that they were item-specific. Repeated word generation in response to aural cues also showed anatomically selective activity reductions within the left frontal and inferior temporal regions, indicating that these activity reductions were not dependent on the perceptual modality of the cue. The auditory cortex showed minimal repetition-related reductions. The presence of activity within left frontal regions that decreases as a function of item repetition for both visual and auditory cues suggests that these reductions may underlie an amodal repetition-priming effect existing at processing stages involving lexical/semantic search and access. The surprising finding that activity reductions in the inferior temporal cortex can be linked to repetition of either visual or auditory cues further suggests that these regions may be modulated in a top-down fashion during repetition priming, independent of (or in parallel with) stimulus-driven perceptual processes. Taken collectively, the data converge on a neural correlate of lexical/semantic priming. Amodal lexical/semantic processes, which may be triggered initially by modality-specific cues, proceed via an interaction between frontal and posterior brain regions. These interdependent regions show activity reductions that correlate with facilitated task performance when items are repeated. PMID- 10686184 TI - The natural history of multiple sclerosis: a geographically based study: 8: familial multiple sclerosis. AB - We have examined the demographics and long-term outcome of 1044 patients with sporadic and familial multiple sclerosis in a population-based cohort from London, Ontario. The mean follow-up was 25 years in duration, and by this time most patients had reached the unambiguous endpoint scores of the Kurtzke disability status scale (DSS), DSS 6, 8 or 10. An affected family member was identified in 19.8% of the total population, and this subgroup was further divided arbitrarily into the following three groups by the type and number of relatives affected: (i) first degree only; (ii) first degree plus others; (iii) second or third degree. The outcome in these groups was compared with that for those patients who, at a mean 25 year follow-up, had no relatives known to be affected. Familial cases closely resembled those remaining sporadic in both demographics and outcome, although onset in the most heavily loaded families was earlier and male/female ratio was greater. The times to DSS 6, 8 and 10 did not differ significantly when sporadic, familial and familial subgroups were compared. These results provide no clinical support for viewing familial multiple sclerosis as distinct from the sporadic form. The observed recurrence rate for siblings in a strictly defined epidemiological sample was 3.5%, much as projected. These results validate the recurrence risks which have previously been derived from age-corrected data for these first-degree relatives. PMID- 10686185 TI - Uses of embryo duplication in humans: embryology and ethics. PMID- 10686186 TI - The welfare of the child: problems of indeterminacy and deontology. PMID- 10686187 TI - Can GnRH agonists act directly on the ovary and contribute to cyst formation? PMID- 10686188 TI - The right to choose your donor: a step towards commercialization or a step towards empowering the patient? PMID- 10686189 TI - Gamete donation: when does consent become irrevocable? PMID- 10686190 TI - A double-blind, randomized study to compare recombinant human follicle stimulating hormone (FSH; Gonal-F) with highly purified urinary FSH (Metrodin) HP) in women undergoing assisted reproductive techniques including intracytoplasmic sperm injection. The French Multicentre Trialists. AB - This prospective, double-blind, randomized, multicentre study compared the efficacy and safety of recombinant human follicle stimulating hormone (r-hFSH; Gonal-F((R))) versus highly purified urinary FSH (u-hFSH HP; Metrodin((R)) HP) in women undergoing ovarian stimulation for in-vitro fertilization, including intracytoplasmic sperm injection. A total of 278 patients began a long gonadotrophin-releasing hormone agonist protocol, then 139 received r-hFSH and 139 u-hFSH HP, 150 IU/day administered s.c., for the first 6 days of treatment. On day 7, the dose was adjusted, if necessary, according to ovarian response. Human chorionic gonadotrophin (HCG, 10 000 IU, s.c.) was administered once there was more than one follicle 18 mm in diameter and two others >/=16 mm. Oocyte retrieval was performed 36-38 h after HCG injection: 128 patients (92%) receiving r-hFSH and 113 (81%) receiving u-hFSH HP had at least one oocyte retrieved. Among patients receiving r-hFSH, there was a significantly higher mean (+/- SD) number of oocytes retrieved (11.0 +/- 5.9 versus 8.8 +/- 4.8 with u-hFSH HP; P = 0. 002) and mean number of embryos obtained (5.1 +/- 3.7 versus 3.5 +/- 2.9 with u-hFSH HP; P = 0.0001). With r-hFSH, significantly fewer FSH treatment days (11.7 +/- 1.9 versus 14.5 +/- 3.3) and 75 IU ampoules (27.6 +/- 10.2 versus 40.7 +/- 13.6) were required than with u-hFSH HP (P = 0.0001). Embryo replacement on day 2-3 after oocyte retrieval resulted in 36 liveborn children in the Gonal-F((R)) group and 33 in the Metrodin HP((R)) group (not significant). There were seven cases (5.0%) of ovarian hyperstimulation syndrome in the r-hFSH group and three (2.2%), in the u-hFSH HP group (not significant). It is concluded that r-hFSH is more effective than u-hFSH in inducing multiple follicular development. PMID- 10686191 TI - Ovarian stimulation with HMG: results of a prospective randomized phase III European study comparing the luteinizing hormone-releasing hormone (LHRH) antagonist cetrorelix and the LHRH-agonist buserelin. European Cetrorelix Study Group. AB - In this prospective and randomized study, 188 patients received the luteinizing hormone-releasing hormone (LHRH) antagonist cetrorelix, and 85 patients the LHRH agonist buserelin to prevent endogenous luteinizing hormone (LH) surges during ovarian stimulation in in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles. Ultimately, 181 patients (96.3%) in the cetrorelix group, and 77 (90.6%) in the buserelin group, reached the day of the human chorionic gonadotrophin (HCG) injection. The mean number of human menopausal gonadotrophin (HMG) ampoules administered and the mean number of stimulation days with HMG were significantly less in the cetrorelix group than in the buserelin group (P < 0.01). A rise in LH and progesterone concentrations was observed in three of the 188 patients (1.6%) who received cetrorelix. On the day of the HCG administration, more follicles of a small diameter (11-14 mm) were observed in the buserelin group than in the cetrorelix group (P = 0. 02) and the mean serum oestradiol concentration was significantly higher in patients who received buserelin than in those who received cetrorelix (P < 0.01). Similar results were observed in fertilization, cleavage and pregnancy rates in the two groups. In conclusion, the use of the LHRH antagonists might be considered more advantageous because of the short-term application needed to inhibit gonadotrophin secretion, so allowing a reduction in the treatment time in a clinically significant manner. PMID- 10686192 TI - Peripheral blood concentrations of inhibin B are elevated during gonadotrophin stimulation in patients who later develop ovarian OHSS and inhibin A concentrations are elevated after OHSS onset. AB - Ovarian hyperstimulation syndrome (OHSS) is a serious side-effect of controlled ovarian stimulation. Inhibin A and inhibin B, as putative predictors of OHSS development in the same stimulation cycle, were evaluated. A cohort of 428 in vitro fertilization (IVF) patients was followed. Fifteen patients with severe OHSS were compared with matched (age, follicle number) controls. Serum samples were obtained at five time points from the start of ovarian stimulation until >/= 3 days post-embryo transfer and analysed with specific enzyme-linked immunosorbent assays. Inhibin A in the OHSS group showed a continuous increase with a significant elevation 3 days prior to oocyte aspiration (ASP-3) and onwards. Maximal concentrations were detected at embryo transfer and the concentrations remained high at >/= 3 days post-embryo transfer. Inhibin A concentrations in the control group showed a transient elevation (significant increase at ASP and embryo transfer). Inhibin A in the OHSS group was significantly higher than in controls only at the time point where OHSS had developed (>/= 3 days post-embryo transfer), and declined during OHSS treatment. Overall, there was a positive correlation between the number of follicles and inhibin A concentrations at ASP-3 until embryo transfer in the control group but not in the OHSS group. The concentrations of inhibin B in both groups increased from the start of ovarian stimulation, with peak values at ASP-3, and then a decline. Inhibin B was significantly higher in OHSS patients at ASP-3 and at ASP. Inhibin B at ASP-3 was correlated with the total number of follicles in both the OHSS group and the control group. PMID- 10686193 TI - Predictive value of serum and follicular fluid leptin concentrations during assisted reproductive cycles in normal women and in women with the polycystic ovarian syndrome. AB - Leptin is an adipocyte-derived hormone which plays a central role in the regulation of body weight and energy homeostasis and in signalling to the brain that adequate energy stores are available for reproduction. Although leptin may affect reproduction by regulating the hypothalamic-pituitary-gonadal axis, recent in-vitro observations indicate that leptin may also have direct intra-ovarian actions. Leptin concentrations were measured in women who succeeded in becoming pregnant within three cycles of in-vitro fertilization (IVF) or gamete intra fallopian transfer (n = 53), in women who failed to become pregnant within three cycles (n = 50), and in women with polycystic ovarian syndrome (PCOS) (n = 22). It was found that lower follicular fluid leptin concentrations were a marker of assisted reproduction treatment success in normal women. Women with PCOS had higher leptin concentrations than women without such a diagnosis, but this was due to their higher body mass index (BMI). After adjustment for age and BMI, women with PCOS who became pregnant tended to have lower mean follicular fluid leptin concentrations than women with PCOS who did not succeed at becoming pregnant. Further studies exploiting the strengths of the IVF model are needed to assess whether the prognostic role for follicular fluid leptin in human reproduction is independent of other factors, and to elucidate the underlying mechanisms. PMID- 10686194 TI - Increased prevalence of thyroid antibodies in euthyroid women with a history of recurrent in-vitro fertilization failure. AB - This study was undertaken to evaluate whether the presence of thyroid antibodies in euthyroid women is associated with an adverse outcome in an in-vitro fertilization (IVF)-embryo transfer programme. In 24 women (study group: mean age +/- SD: 31.5 +/- 4.4 years) who failed to conceive after having three or more cycles of IVF and embryo transfer, serum concentrations of thyroglobulin (TG), thyroid peroxidase antibodies (TPO) and anticardiolipin antibodies (IgG and IgM) were measured using commercially available kits. The control group comprised 24 consecutive patients without endocrine dysfunction (mean age +/- SD: 30.3 +/- 4.1 years) seeking infertility treatment in our department of assisted reproduction. All patients in both the study and the control groups were determined to be euthyroid by demonstrating normal concentrations of thyroid-stimulating hormone (TSH). In the study and control groups respectively, 13 and two patients demonstrated positive titres of TG, TPO or both thyroid antibodies (Fisher's exact test: P = 0.002). Mean serum concentrations of TG were significantly increased in the study group compared to the control subjects (156 +/- 167 IU/ml versus 33.5 +/- 32.0 IU/ml; U-test: P = 0.009). Serum concentrations of TPO and anticardiolipin antibodies were similar in both groups. Our investigations revealed that thyroid antibodies might be independent markers for reproductive failure in an IVF-embryo transfer programme. PMID- 10686195 TI - Serum concentrations of dimeric inhibins, activin A, gonadotrophins and ovarian steroids during the menstrual cycle in older women. AB - The transition from regular ovarian cyclicity to menopause is associated with a rise in the circulating concentrations of follicle stimulating hormone (FSH), despite the maintenance of serum oestradiol concentrations during the perimenopause. The aim of this study was to compare the pattern of secretion of dimeric inhibins, activin A, gonadotrophins and steroids in regularly cycling women of 40-50 years with normal and raised early follicular phase serum FSH concentrations and young women (25-33 years) during the menstrual cycle. Blood samples were taken prospectively almost daily throughout the menstrual cycle. Women recruited were classified into three groups: (i) older women with normal FSH [(ON-FSH), day 3 FSH <8 mIU/ml, n = 10]; (ii) older women with raised FSH [(R FSH), day 3 FSH >8 mIU/ml, n = 6] and (iii) young normal FSH (YN-FSH) women, age 25-32 years (n = 6). Cyclic patterns of serum inhibins and activin A were similar in the ON-FSH and YN-FSH groups. The R-FSH group had significantly lower concentrations of inhibin A prior to the luteinizing hormone (LH) surge and in the mid-luteal phase and lower concentrations of inhibin B in the early follicular phase compared with the ON-FSH group. Serum concentrations of activin A, progesterone and oestradiol were similar in all three groups. It is concluded from this study that the rise in early follicular phase serum FSH in older women is associated with a decrease in circulating concentrations of inhibin B in the early follicular phase. However, lower circulating concentrations of inhibin A in the luteal phase of the R-FSH group may also contribute to the rise in early follicular phase FSH concentrations during the menstrual cycle, although further studies with larger numbers are required to confirm this observation. PMID- 10686196 TI - Cell proliferation and vascular morphology in the marmoset corpus luteum. AB - Luteal formation is associated with angiogenesis and low progesterone production. Maximal mid-luteal phase progesterone production is concurrent with extensive vascularization, and luteolysis occurs when steroidogenesis decreases. Angiogenic cell proliferation and vascular changes have not been examined in the marmoset. The aim of this study was to examine vascular morphology throughout the luteal phase by identifying: (i) von Willebrand factor VIII antigen (vW)-immunopositive endothelial cells; (ii) Ki67-positive proliferating cells; and (iii) bromodeoxyuridine-positive proliferating cells. Marmoset corpora lutea were examined throughout the cycle, and natural regression was compared with induced luteolysis after administration of a prostaglandin F(2alpha) analogue or gonadotrophin-releasing hormone (GnRH) antagonist. Steroidogenic and endothelial cells were positive for proliferation markers. Endothelial cell proliferation was highest during luteal formation, then decreased and remained low during the luteal phase and functional regression, however endothelial cell proliferation increased during structural regression. Endothelial cell proliferation was unchanged by induced regression. The area of vW immunostaining was highest during luteal formation, decreased thereafter and remained constant during the luteal phase and regression. Distribution of immunostaining indicated the presence of an extensive capillary network, but during structural regression the numbers of capillaries decreased and numbers of microvessels increased. These results suggest that vascular changes are concurrent with changes in the functional status of the marmoset corpus luteum. PMID- 10686197 TI - Rescue of oocytes from antral follicles of cryopreserved mouse ovaries: competence to undergo maturation, embryogenesis, and development to term. AB - Only primordial and primary follicles of frozen-thawed mouse ovaries survive after grafting to the ovarian bursa; large secondary follicles and antral follicles together with the oocytes contained in them degenerate. This study was undertaken to determine whether fully grown oocytes isolated from the antral follicles of frozen-thawed mouse ovaries are viable and can be rescued to undergo maturation, fertilization, and embryo development in vitro. Ovaries were cryopreserved after removal from 22-day-old (C57BL/6J x SJL/J)F(1) mice, with or without prior priming with equine chorionic gonadotrophin, and fresh non-frozen ovaries were used as controls. Only cumulus cell-denuded oocytes were recovered from frozen unprimed ovaries while both cumulus cell-enclosed and denuded oocytes were retrieved from frozen primed ovaries. Oocytes from both groups of frozen thawed ovaries were able to undergo maturation, fertilization, and development to the blastocyst stage in vitro, though at lower percentages than oocytes from control unfrozen ovaries. Moreover, 19% of 2-cell stage embryos derived from frozen-thawed primed ovaries, compared with 42% of embryos derived from control primed ovaries, developed to term after transfer to pseudopregnant foster mothers (not significantly different). Therefore, fully grown oocytes in antral follicles survive the cryopreservation protocol, as demonstrated by maturation, fertilization and embryo development in vitro, and development to term after embryo transfer. PMID- 10686198 TI - Transplantation of cultured explants of human endometrium into nude mice. AB - The objective of this study was to analyse the histological and immunohistochemical characteristics of cultured explants of human endometrium transplanted into nude mice. Biopsies of eutopic endometrium were taken from six patients during laparoscopic surgery and classified according to the phase of the cycle. All the explants were cultured with oestrogen and progesterone for 24 h before transplantation into 15 mice. Four mice were grafted with explants of menstrual endometrium, four with explants of late proliferative endometrium, and seven with explants of late secretory-premenstrual endometrium. Typical endometrial glands and stroma were observed in 87% of cases 3 weeks after the transplantation. All the grafts revealed histological characteristics of the proliferative phase, even when the endometrial biopsy was taken during the late secretory phase. Immunohistochemical studies revealed that the proliferation index was high, whatever the menstrual phase of the endometrial biopsy. An extensive vascular network developed at the interface between the graft and the surrounding tissue. Vascular endothelial growth factor (VEGF) positive stained cells were observed in all grafts, the VEGF score being significantly higher in epithelial cells than in stromal cells. In conclusion, human endometrial explants, cultured for 24 h, could be successfully transplanted into nude mice. Immunohistochemical studies proved that human cultured endometrial tissue could not only survive in nude mice but could also become very active and develop characteristics different from the tissue of origin. An active vascular network is a necessary condition for the survival of the graft and may be explained by the high VEGF content. PMID- 10686199 TI - Production of inhibin forms by the fetal membranes, decidua, placenta and fetus at parturition. AB - Inhibins are regulators of paracrine and endocrine function during pregnancy, but their intrauterine sites of secretion are not well established. In amniotic fluid, inhibin A-, inhibin B- and inhibin pro-alphaC-containing isoforms were present in high concentrations, whereas in maternal serum, inhibin A and pro alphaC forms were present in high amounts, with low concentrations of inhibin B. In fetal cord serum, inhibin pro-alphaC was present in all samples, inhibin B was detectable in male but not female fetuses, with no detectable inhibin A in either sex. From cultured explants, both inhibin A and B were secreted by chorion laeve, whereas only inhibin A was secreted by placenta, with both tissues secreting inhibin pro-alphaC. Only low concentrations of both dimeric inhibins and pro alphaC forms were secreted by decidua parietalis and amnion. The dual perfused placental cotyledon secreted both inhibin A and pro-alphaC into maternal perfusate, but only inhibin pro-alphaC into the fetal circulation and less than to the maternal side. We conclude that trophoblast is the predominant source of dimeric inhibins, but with markedly different secretion depending on its intrauterine location. There was a significant decrease in inhibin A and pro alphaC in amniotic fluid collected at term active labour compared to elective Caesarean section (P < 0.001). This may reflect a local change in inhibin/activin processing at labour, likely in chorion laeve trophoblast cells, which may be important in the paracrine control of the feto-maternal communication required to maintain pregnancy and initiate labour. PMID- 10686200 TI - Fluorescence diagnosis of endometriosis on the chorioallantoic membrane using 5 aminolaevulinic acid. AB - The chorioallantoic membrane (CAM) is a useful model for the fluorescence diagnosis of experimentally induced endometriosis. In our experimental setup 75.7% of the histologically examined tissue preparations were viable and only 24.3% showed signs of necrosis on the CAM after various periods of incubation. Best results were obtained when grafting to the CAM was performed between days 7 and 9 and when implants were left on the CAM for 3-5 days (P < 0.05). We were able to demonstrate that 5-aminolaevulinic acid (ALA) is stored selectively in ectopic endometrium. The subsequent fluorescence of the endometrium shows a rapid increase that reaches a peak after 10-14 h which can be clearly differentiated from the weaker fluorescence of grafted normal peritoneum and fimbriae (P < 0.01). PMID- 10686201 TI - Reproductive features in women developing ovarian granulosa cell tumour at a fertile age. AB - Ovarian granulosa cell tumour (GCT) is a rare malignancy, which has been linked to both infertility and infertility treatment with ovulation inducers. The reproductive features were analysed of 146 women with GCT diagnosed between 1956 and 1996. During the study period no changes were found in the mean age (53 years), menopausal status (59% postmenopausal), parity (32% nulliparous) or tumour size or stage at diagnosis. The clinical features in women with GCT at fertile age were compared with GCT diagnosed later in life and to population based data. Nulliparity (50%) and history of infertility (22%) were more frequent if the tumour occurred at fertile age (n = 50). Of the 12 infertile cases, seven had anovulatory infertility (58%); 11 occurred during the era of ovulation inducers, but only five had used these drugs (clomiphene citrate in five patients, gonadotrophins in two, and tamoxifen in one patient) and no patient had undergone in-vitro fertilization. Endometrial hyperplasia was associated with GCT at all ages, while endometrial cancer was found solely after the age of 45 years. In conclusion, GCT at fertile age is associated with nulliparity and with a clinical presentation of anovulatory infertility, while GCT later in life is associated with a more normal average fertility pattern and with occurrence of endometrial cancer. PMID- 10686202 TI - Repetitive complete hydatidiform mole can be biparental in origin and either male or female. AB - Complete hydatidiform mole (CHM) is an abnormality in pregnancy due to a diploid conception which is generally androgenetic in origin, i. e. all 46 chromosomes are paternally derived. We have examined the genetic origin of repetitive hydatidiform moles in a patient having three CHM by two different partners, and no normal pregnancies. Using fluorescent microsatellite genotyping, we have shown all three CHM to be biparental, rather than androgenetic, in origin. Examination of informative markers for each homologous pair of chromosomes, in two of the CHM, failed to reveal any evidence of unipaternal disomy, suggesting that the molar phenotype might result from disruption of normal imprinting patterns due to a defect in the maternal genome. It has been suggested that intracytoplasmic sperm injection (ICSI), followed by selection of male embryos, can prevent repetitive CHM; but examination of sex chromosome-specific sequences in the three CHM described here, showed that, while two were female, the first CHM was male. Selection of male embryos is therefore unlikely to prevent repetitive CHM in this patient. Our results suggest that the genetic origin of repetitive CHM should be determined prior to in-vitro fertilization (IVF) and that current strategies for the prevention of repetitive CHM may not be appropriate where the CHM are of biparental origin. PMID- 10686203 TI - Menopause and risk of non-fatal acute myocardial infarction: an Italian case control study and a review of the literature. AB - The relationship between menopause and non-fatal acute myocardial infarction (AMI) was considered by analysing data from a case-control study conducted in Italy between 1983 and 1992. Cases were 429 women, below age 75 years, with a first episode of non-fatal AMI, admitted to 30 coronary care units; controls were 863 women admitted to the same network of hospitals for acute diseases other than cardiovascular, neoplastic, or hormone-related. Postmenopausal women were not at higher risk of AMI than pre/perimenopausal women, after adjustment for age and other selected covariates [multivariate odds ratio (OR) 0.99]. With reference to age at menopause, compared with women reporting menopause when <45 years, the multivariate OR were 1.54 for those aged 45-49 at menopause, 1.36 for those aged 50-52 years, and 0.97 for those aged >/=53, in the absence of any trend in risk. No meaningful relationship emerged with time since menopause (OR 0.85 for <10 years since menopause). The results were similar in women aged <60 and >/=60 years at AMI. Although the present study does not support a substantial relationship between menopause and non-fatal AMI, the overall epidemiological evidence is compatible with a moderate association. PMID- 10686204 TI - Incidence of cancer in children born after in-vitro fertilization. AB - Evaluation of the long-term health of children born using in-vitro fertilization (IVF) provides important information to clinicians and consumers. Until very recently, there have been no published data on the incidence of cancer in children conceived as a result of IVF, despite a number of case reports of neuroblastoma in children conceived using fertility drugs. This study used a record-linkage cohort design to investigate the incidence of cancer in children born after IVF. The study included all conceptions using assisted reproductive technologies between 1979 and 1995 at two clinics in Victoria, Australia that resulted in a live birth. Data on births were linked with a population-based cancer registry to determine the number of cases of cancer that occurred. The standardized incidence ratio (SIR) was calculated by comparing the observed number of cases to the expected number of cases. The final cohort included 5249 births. The median length of follow-up was 3 years, 9 months (range 0-15 years). In all, 4.33 cases of cancer were expected and six were observed, giving a SIR of 1.39 (95% CI 0.62-3.09). This study found that children conceived using IVF and related procedures did not have a significantly increased incidence of cancer in comparison to the general population. PMID- 10686205 TI - Hydatidiform mole coexistent with a twin live fetus: a national collaborative study in Japan. AB - A national collaborative study was conducted in Japan to evaluate the clinical course and the sequelae of patients with hydatidiform mole coexistent with twin live fetus (HMTF). Seventy-two cases of HMTF were diagnosed based on gross appearance and histopathological criteria. In 18 cases, the molar parts were cytogenetically confirmed to be of androgenetic origin (complete mole). The overall incidence of persistent trophoblastic tumour (PTT) in patients with HMTF was 30.6%, and it increased to 50.0% in the 18 patients with proven androgenetic complete mole coexistent with twin live fetus (CHMTF). Among these patients, the mean gestational age at termination of pregnancy or delivery in those who developed PTT (n = 9) and those who did not (n = 9) were 20.6 and 19.4 weeks respectively. The incidence of severe maternal complications was significantly higher in patients who subsequently developed PTT (P < 0.05). The rate of subsequent development of PTT in patients with CHMTF was found to be considerably higher than in a previous study of patients with single complete mole (50 and 12.5% respectively). However, since the risk of malignancy is unchanged with advancement of gestational age, continued pregnancy may be allowed in patients with HMTF provided that severe maternal complications are controlled and fetal karyotype and development are normal. PMID- 10686206 TI - Polycystic ovaries and recurrent miscarriage--a reappraisal. AB - The prevalence of polycystic ovaries (PCO) was established amongst 2199 consecutive women (median age 33 years; range 19-46) with a history of recurrent miscarriage (median 3; 3-14). A diagnosis of PCO was made if the ovarian volume was enlarged (>9 ml), there were >/=10 cysts of 2-8 mm in diameter in one plane and there was increased density of the stroma. In a cohort study, the prospective pregnancy outcome of 486 of the women scanned who were antiphospholipid antibody negative and who received no pharmacological treatment during their next pregnancy was studied. The prevalence of PCO was 40.7% (895/2199). The livebirth rate was similar amongst women with PCO (60.9%; 142/233) compared to that amongst women with normal ovarian morphology (58.5%; 148/253; not significant). Neither an elevated serum luteinizing hormone concentration (>10 IU/l) nor an elevated serum testosterone concentration (>3 nmol/l) was associated with an increased miscarriage rate. Polycystic ovarian morphology is not predictive of pregnancy loss amongst ovulatory women with recurrent miscarriage conceiving spontaneously. The search for a specific endocrine abnormality that can divide women with PCO into those with a good and those with a poorer prognosis for a future successful pregnancy continues. PMID- 10686207 TI - Embryo transfer under ultrasound guidance improves pregnancy rates after in-vitro fertilization. AB - Between October 1998 and January 1999, we examined the influence of ultrasound guidance in embryo transfer on pregnancy rate in 362 patients from our in-vitro fertilization (IVF)-embryo transfer programme. These patients were prospectively randomized into two groups: 182 had ultrasound-guided embryo replacement, and 180 had clinical touch embryo transfer. There were no statistically significant differences between the two groups with respect to age, cause of infertility and in the characteristics of the IVF cycle. The pregnancy rate was significantly higher among the ultrasound-guided embryo transfer group (50%) compared with the clinical touch group (33.7%) (P < 0.002). Furthermore, there was also a significant increase in the implantation rate: 25.3% in the ultrasound group compared with 18.1% in the clinical touch group (P < 0.05). In conclusion, ultrasound assistance in embryo transfer significantly improved pregnancy and implantation rates in IVF. PMID- 10686208 TI - Intrauterine donor insemination in single women and lesbian couples: a comparative study of pregnancy rates. AB - The outcome of intrauterine donor insemination (IUI-DI) with frozen spermatozoa was analysed retrospectively in 675 cycles in single women (n = 122; 536 cycles) and lesbian (n = 35; 139 cycles) couples. The lesbian patients were younger at the initiation of treatment (mean 34.5 years; range 26-44) than the single women (mean 38.5; range 29-47) (P = 0.005). Clinical pregnancy rate was 36% in single women and 57% in lesbians (P < 0.05), the cumulative pregnancy rate after six cycles being 47% and 70% respectively, although the outcome was similar when related to age. The miscarriage rate was higher (35%) in single women than in lesbians (15%; P < 0.05), the rate being independent of maternal age. There were no apparent differences seen between the two groups with respect to the possible effect of parity, duration of infertility, causes of infertility and type of treatment at initiation of treatment; the sole exception was that the age of lesbian women was statistically significantly younger than that of single women (P < 0.005). When corrected for age, the pregnancy rates and complications were lower and higher respectively in single women but these differences did not reach statistical significance. However, the disparity between the treatment outcomes of single women and lesbian patients of similar ages may also reflect the fact that single women are likely to have failed to conceive for a period of time prior to referral to a specialist centre for treatment. PMID- 10686209 TI - Successful twin pregnancy in a dual-transplant couple resulting from in-vitro fertilization and intracytoplasmic sperm injection: case report. AB - There are numerous reports of successful pregnancy following liver transplantation. Little information is available regarding the incidence and management of infertility in transplant recipients, particularly the use of artificial reproductive technologies. We present a case of a successful twin pregnancy resulting from in-vitro fertilization with intracytoplasmic sperm injection (IVF/ICSI) in a liver transplant recipient, whose partner was a renal transplant recipient with severe oligozoospermia. With careful evaluation and monitoring, and the involvement of appropriate consultants, artificial reproductive technologies can be safely used in transplant recipient couples experiencing infertility. PMID- 10686210 TI - Single monthly administration of the anti-progestagen Org 31710 in users of the 75 microg desogestrel progestagen-only pill: effects on pituitary-ovarian activity. AB - Endocrine and ultrasound effects were studied of an intermittent (every 28 days) oral administration of 150 mg of the anti-progestagen Org 31710 during the continued daily use of 75 microg desogestrel (DSG) for progestagen-only contraception. A randomized, double-blind, placebo-controlled two-centre study was conducted in 50 healthy volunteers. Serum luteinizing hormone (LH), follicle stimulating hormone (FSH), oestradiol and progesterone concentrations, and follicle number and size were studied, as well as endometrial thickness, which was assessed by transvaginal sonography at least twice weekly during a single medication cycle (cycle 3-5). Forty-eight women were evaluated (Org 31710, n = 25; placebo, n = 23). Seven ovulations were observed in the treated group versus none in the placebo group. LH concentrations were higher on days 9 and 11 and oestradiol concentrations lower on day 3 in the treated group, irrespective of whether ovulation occurred. No parameter could predict ovulation. Endometrial thickness was greater on cycle days 7-13 and 19 in the treated group. However, within the Org 31710 group, no significant differences were found in volunteers who did or did not ovulate. Observed differences may be attributed to a competitive effect of Org 31710 with progestagen-induced suppression of the pituitary-ovarian axis, altered oestradiol feedback mechanisms, and/or altered receptor availability. PMID- 10686211 TI - Potential impact of hormonal male contraception: cross-cultural implications for development of novel preparations. AB - The prospect of a hormonal male contraceptive is no longer distant. Data on the potential impact of this improvement in contraceptive provision, however, is limited, particularly between different cultures. We have therefore carried out a multi-centre study to assess men's attitudes to proposed novel hormonal methods. Questionnaire-based structured interviews were administered to men in Edinburgh, Cape Town, Shanghai and Hong Kong. Approximately 450 men were interviewed in Edinburgh, Shanghai and Hong Kong, and a slightly larger group (n = 493) in Cape Town to give samples (n > 150) of black, coloured and white men. Knowledge of existing male and female methods of contraception was high in all centres and groups. The majority of men welcomed a new hormonal method of contraception, 44 83% stating that they would use a male contraceptive pill. Overall, a pill was more acceptable than an injectable form (most popularly given at 3-6 month intervals); long-acting implants were least so except in Shanghai. Familiarity with comparable female methods appeared to influence acceptability, for both oral and injectable methods. Hong Kong was the only centre where a male method (condom) was currently the most commonly used; men there appeared to rate the convenience of condoms highly while being least likely to think that they provided effective protection against pregnancy compared to other centres, and were least enthusiastic about novel male methods. The acceptability of potential male hormonal methods of contraception was high in some groups but showed wide variability, determining factors including cultural background and current contraceptive usage. These results suggest that the emerging emphasis that men should have greater involvement in family planning will be substantiated when appropriate contraceptive methods become available. PMID- 10686212 TI - Would women trust their partners to use a male pill? AB - Despite a renewed interest in the development of hormonal contraceptives for men, many discussions about the potential acceptability of a 'male pill' end by speculating whether women would trust their partners to use the method reliably. To determine the views of women, we undertook a survey of 1894 women attending family planning clinics in Scotland (450), China (900) and South Africa (544). In all centres over 65% of women thought that the responsibility for contraception falls too much on women. More than 90% in South Africa and Scotland thought that a 'male pill' was a good idea, with Chinese women (71% in Hong Kong and 87% in Shanghai) only slightly less positive. Only 13% of the total sample did not think that hormonal male contraception was a good idea and only 36 women (2% of the total) said that they would not trust their partner to use it. 78% of Scottish women, 71% of Shanghai women, and 78% of white women and 40% of black and coloured women in Cape Town thought that they would use the method. This survey should dispel the myth that women would not trust their partners to use a 'male pill' reliably and illustrates the potential market for the method. PMID- 10686213 TI - Intrauterine polyps--a cause of unscheduled bleeding in women using the levonorgestrel intrauterine system: case report. AB - The levonorgestrel intrauterine releasing system is a contraceptive that has been shown to reduce menstrual blood loss dramatically. Breakthrough bleeding, however, is a relatively common occurrence as with all methods of progestogen only contraception and this limits its acceptability for women. Amenorrhoea can be achieved in the majority of women within 12 months of insertion. Any new pattern of bleeding after amenorrhoea or a persistence of heavy bleeding may be due to co-existing intrauterine pathology such as endometrial polyps. The use of out-patient techniques such as hysteroscopy and saline infusion sonography are indicated in these instances to exclude other intrauterine pathology. PMID- 10686214 TI - Prospective comparative study between microsurgical and conventional testicular sperm extraction in non-obstructive azoospermia: follow-up by serial ultrasound examinations. AB - The value of testicular sperm extraction (TESE) by microdissection was evaluated according to its physiological consequences compared with open, classic surgical biopsy in the same patient. A total of 100 patients with non-obstructive azoospermia and bilateral identical testicular histology underwent bilateral diagnostic TESE via the conventional method on one side and the microsurgical method on the other side. The spermatozoa recovery rate by microdissection TESE was significantly higher than by conventional TESE (47 and 30% respectively; P < 0.05). In order to assess the safety of this new procedure, 60 patients were followed-up ultrasonographically for 1, 3 and 6 months. Acute and chronic complications were significantly lower in the microsurgical side compared with the conventional side (15 and 58.3% respectively and 3 and 30% respectively; P < 0.05). Segmental devascularization was detected in seven testes operated on conventionally, and in two testes operated on microsurgically. However, permanent devascularization could not be detected in any patient after 6 months. These findings suggest that microdissection TESE is not devoid of complications, but that it is relatively safer than the conventional technique and improves sperm yield significantly in patients with non-obstructive azoospermia. PMID- 10686215 TI - Sperm chromatin packaging as an indicator of in-vitro fertilization rates. AB - The development of a sequential diagnostic schedule for patients consulting for infertility disturbances would be an ideal method of approach for clinicians in the absence of an aetiological or pathophysiological diagnosis. Since sperm morphology recorded by strict criteria has often been correlated with fertilization failure, the present study aimed to evaluate the relationship between normal morphology as well as in-vitro fertilization (IVF) rates, with chromatin staining among fertile and subfertile men. Two semen smears were prepared from each specimen obtained from 72 men to record normal morphology and chromatin packaging as recorded by chromomycin A(3) (CMA(3)) staining. Following the semen analyses, the 72 men were divided into the two morphological groups, namely <4% and >4% normal forms. Significantly different percentages of CMA(3) staining (mean +/- SE) were recorded between the two morphological groups, namely 65.9% +/- 3.5 and 44.5% +/- 1.7 (P +/- 0.001). A highly negative significant correlation existed between percentage of normal morphology as recorded by strict criteria and CMA(3) staining. A highly significant and positive correlation was recorded for normal morphology and IVF rates (r +/- 0.45, P +/- 0.0001). A significant negative correlation (r +/- -0.51, P +/- 0.0001) existed between CMA(3) values and IVF rates. The discriminating power of nuclear maturity, as recorded by CMA(3) staining, to identify abnormal morphology values and poor IVF rates was calculated with receiver operator characteristic (ROC) analyses. The areas under the ROC curves were 0.86 for sperm morphology and 0. 74 for IVF rates. The calculated threshold values for CMA(3) staining to distinguish between morphology groups were 48 and 50% for IVF. Chromatin packaging assessment is a valuable addition to the sequential diagnostic programme in an assisted reproductive arena. PMID- 10686216 TI - Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica particles at different g forces. AB - The effects of density gradient centrifugation through silane-coated silica particles (PureSperm) using 100, 200, 300 and 500 g on bacterial contamination of sperm samples and recovery of motile spermatozoa from sperm samples were investigated with conventional culturing techniques and microscopic visual assessment. The recovery of motile spermatozoa was variable and was not improved using 500 g compared to the recommended 300 g. The bacterial contamination was highly decreased by gradient centrifugation through PureSperm and was almost abolished when strict aseptic techniques were used, with changes to sterile Pasteur pipettes and tubes prior to washing procedures. PMID- 10686217 TI - The development of a continuous quality control programme for strict sperm morphology among sub-Saharan African laboratories. AB - Inter-technician and between-laboratory differences, especially during the evaluation of sperm morphology, have been a major cause of concern. The study aimed to develop an intensive training programme with intervals of continuous quality control assessments for sperm morphology. Twenty andrology laboratories from sub-Saharan Africa were invited to participate in a World Health Organization Special Programme of Research, Development and Research Training in Human Reproduction semenology workshop. Following intensive training in strict sperm morphololgy evaluation, a continuous quality control programme was introduced on a quarterly basis. At baseline, the mean (+/- SD) percentage difference reported between the participants and the reference laboratory reading was 33.50 +/- 11%. After training, the mean percentage difference had decreased to 14.32 +/- 5% at 3 months and to 5.00 +/- 5% at 6 months. Pairwise comparison of the differences at each evaluation time revealed the following: Baseline differences (pre-training) differed significantly from the differences at 3 months (P = 0.0002) as well as at 6 months after training (P = 0.007). The differences at 6 months did not differ significantly from those at 3 months (P = 0.27). Training of andrology technicians as well as continuous proficiency testing can be conducted on a national and international level with the support of a referring laboratory. Global quality control measurements in andrology laboratories should become mandatory, since these results indicate that continuous quality control for laboratory technicians can be highly successful. PMID- 10686218 TI - Oestradiol enhances testosterone-induced suppression of human spermatogenesis. AB - The aim of this study was to determine for the first time in humans, the efficacy of adding a low dose oestradiol to a suboptimally suppressive testosterone dose in a depot hormonal regimen to suppress spermatogenesis in healthy eugonadal men. Twenty-six healthy men were randomized into groups that were treated by a single subdermal implantation of either 600 mg testosterone alone (T; n = 11) or together with 10 mg (TE10, n = 7) or 20 mg (TE20, n = 8) oestradiol. Administration of oestradiol produced a dose-dependent increase in peak plasma oestradiol at 1 month and prolonged suppression of plasma LH and FSH leading to significantly enhanced suppression of sperm output. Despite the augmented spermatogenic suppression, there was no significant difference in the proportions achieving azoospermia (6/26, 23%) or severe oligozoospermia (<1 or <3 x 10(6) spermatozoa per ml, 7/26, 27%) and overall these proportions were inadequate to provide reliable contraception according to the standards identified in World Health Organization male contraceptive efficacy studies. Total and free testosterone remained within the eugonadal reference range for young men throughout the study. While the lower oestradiol dosage had minimal spermatogenic suppression effects, the higher dose produced dose-limiting adverse effects of androgen deficiency and/or oestrogen excess between the fourth and sixth month of the study. This appeared to be due to the unexpectedly prolonged, low concentration of oestradiol release from the oestradiol implants. There were no significant treatment-related changes in body composition, lipids, prostate specific antigen, haematological or biochemical variables. Thus oestradiol has a low therapeutic window and dose-limiting side-effects at dosages that fail to achieve the uniform azoospermia required of an effective male hormonal contraceptive regimen. PMID- 10686219 TI - Results of the American Association of Bioanalysts national proficiency testing programme in andrology. AB - Proficiency testing samples for antisperm antibodies (ASAB), sperm count, morphology and vitality were mailed to participating laboratories. The majority participating utilized Immunobead ASAB procedures (81 versus 14% mixed antiglobulin reaction and 5% 'other'), and there was 95.6 +/- 1.2% agreement on the presence or absence of ASAB. The majority of laboratories utilized manual (79%) versus computer assisted semen analysis (CASA; 15%) methods. Approximately 64% used the haemocytometer and 26% used the Makler counting chambers for manual counts. Coefficients of variation (CV) in sperm counts ranged from 24 to 138%, with CASA displaying lower overall CV (53 +/- 8%) than manual methods (80 +/- 9%). A wide variation in the reports of percent normal morphology was noted (CVs calculated from arc sin transformed means ranged from 15 to 93%). Participants using American Society of Clinical Pathologists (ASCP) criteria reported sperm morphology values that were clustered in the 'normal' range (11 out of 12 samples), while those using strict criteria were clustered in the 'abnormal' range (10 out of 12 samples). Good agreement was observed in sperm vitality (overall mean CV = 18%). These data highlight the urgent need for improvement in overall quality of andrology testing and indicate that practical proficiency testing programmes can be made available on a large scale. PMID- 10686220 TI - Adhesion formation in intubated rabbits increases with high insufflation pressure during endoscopic surgery. AB - The aim of the study was to test the hypothesis that the increase in adhesion formation by CO(2) pneumoperitoneum is caused by mesothelial hypoxaemia. Therefore the effect of the intra-abdominal pressure together with the flow rate upon adhesion formation was evaluated in rabbits following laser and bipolar lesions during endoscopic surgery using humidified CO(2) at 35 +/- 1 degrees C. The intra-abdominal pressure and flow rate were 5 mmHg and 1 l/min in group 1 (n = 5), 5 mmHg and 10 l/min in group 2 (n = 4), 20 mmHg and 1 l/min in group 3 (n = 5) and 20 mmHg and 10 l/min in group 4 (n = 4) respectively. A rapid and reliable intubation method for rabbits was developed to permit high insufflation pressure. By two-way analysis of variance, total adhesion scores following a laser lesion increased with flow rate (P = 0.0003) and insufflation pressure (P = 0.002). Total adhesion scores of bipolar lesions increased with pressure (P = 0.02) but not with flow rate (P = 0.1). The total adhesion scores of laser and bipolar lesions together increased with flow rate (P = 0.005) and with insufflation pressure (P = 0.004). There was no statistical interaction between flow rate and insufflation pressure. In conclusion, the insufflation pressure in endoscopic surgery with CO(2) pneumoperitoneum is a co-factor in adhesion formation, together with desiccation. PMID- 10686221 TI - Laparoscopic creation of a neovagina in a woman with a kidney transplant: case report. AB - The successful use of Vecchietti's technique for creating a neovagina in a case of Rokitansky syndrome with an associated transplanted kidney is reported. The technique is performed by means of a laparoscopic approach, adapted to the special anatomical situation, in order to avoid renal injuries. The accomplishment of a normally functioning neovagina, with no intra-operative or post-operative complications, proves that this technique can also be applied satisfactorily to this type of patient. PMID- 10686222 TI - Flow of cells from polar to mural trophectoderm is polarized in the mouse blastocyst. AB - During growth of the blastocyst there is a net flow of cells from the polar to the mural trophectoderm which is presumed to be radially symmetrical. However, such a pattern of cell movement is inconsistent with findings from a recent clonal analysis. To visualize the overall flow of cells directly, the polar trophectoderm of expanding blastocysts was labelled globally with fluorescent microspheres. Following further growth, the great majority of blastocysts that remained labelled throughout the polar trophectoderm exhibited a polarized rather than radial spread of label into the mural region. This was the case regardless of the labelling technique, whether the blastocysts were grown in utero or in vitro, or had the zona pellucida removed or left on. Intriguingly, where there were two foci of spread of label into the mural trophectoderm rather than one, these were diametrically opposite each other. In further experiments, fluorescent lineage labels were used to distinguish junctional trophectoderm cells with and without an extension onto the blastocoelic surface of the inner cell mass. The location of clones formed following further blastocyst growth provided no evidence that egress of cells from the polar trophectoderm is restricted circumferentially by the presence of junctional cells having an extension. PMID- 10686223 TI - Defective sperm-zona pellucida interaction: a major cause of failure of fertilization in clinical in-vitro fertilization. AB - Sperm-zona pellucida binding and penetration were assessed on the oocytes that failed to fertilize from couples with >/=3 oocytes treated by standard in-vitro fertilization (IVF). There were four groups: fertilization rate 0% (n = 369), 1 25% (n = 194), 26-50% (n = 81) and 51-95% (n = 100). Of the couples with zero fertilization rate 70% had or =50% reduction in baseline scores) were 40% for both the HDRS(28) and the Clinical Global Impressions-Improvement index (score of 1 or 2) and 50% for the Montgomery-Asberg Depression Rating Scale. Symptomatic responses (accompanied by substantial functional improvement) have been largely sustained during long-term follow-up to date. CONCLUSIONS: These open trial results suggest that VNS has antidepressant effects in treatment-resistant depressions. PMID- 10686264 TI - Neurobiology of the obsessive-compulsive spectrum disorders. AB - Advances in obsessive-compulsive disorder (OCD) research have led to increased attention to a range of disorders with possibly overlapping phenomenological and neurobiological features; the so-called OCD spectrum disorders. This article briefly reviews neurobiological data relevant to the construction of an OCD spectrum, including neurochemical, neuroanatomic, genetic, neuroimmunology, and animal studies. OCD and related disorders may be heterogenous conditions, and the neurobiology of many putative OCD spectrum disorders has not been well studied. Nevertheless, a gradual accumulation of neurobiological data has provided a number of exciting, and partially overlapping, approaches to an hypothesized OCD spectrum. PMID- 10686265 TI - Reduced glutamate in the anterior cingulate cortex in depression: an in vivo proton magnetic resonance spectroscopy study. AB - BACKGROUND: Functional imaging studies suggest a specific role of the anterior brain regions in the pathogenesis of major depression. The aim of this study was to evaluate possible neurochemical alterations in the frontomesial cortex in patients with major depressive episode using in vivo proton magnetic resonance spectroscopy ((1)H-MRS). METHODS: Single voxel (1)H-MRS was performed in 19 patients with major depressive episodes and 18 age-matched healthy controls within the anterior cingulate cortex and the parietal white matter. Absolute concentrations were estimated for N-acetyl-aspartate, choline-containing compounds, total creatine, myo-inositol, unresolved glutamate and glutamine (Glx) and glutamate alone (Glu). Voxel composition was analyzed by image segmentation into cerebrospinal fluid (CSF), grey and white matter. RESULTS: MANOVA test for Glx and Glu using age, percent CSF and percent grey matter contribution as covariates yielded a significant group effect within the anterior cingulate due to decrease of Glx in patients (-10.4%, p =.013). Considering only severely depressed patients, both Glx and Glu (-14.3%, p =.03) showed a significant decrease. There was no significant group effect for the neuronal marker NAA, creatine, choline or myo-inositol in either localization. CONCLUSIONS: This study suggests a possible role of altered glutamatergic neurotransmission within the anterior cingulate in the pathogenesis of mood disorders. The otherwise unremarkable findings of major brain metabolites confirms lack of neurodegenerative or membrane metabolic changes in major depression. PMID- 10686266 TI - Repetitive transcranial magnetic stimulation is as effective as electroconvulsive therapy in the treatment of nondelusional major depressive disorder: an open study. AB - BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS), a new method for the stimulation of the central nervous system, is being proposed as a potential new treatment in patients with major depressive disorder (MDD). We tested the hypothesis that rTMS would be as effective as electroconvulsive therapy (ECT) in patients with MDD. METHODS: Forty patients with MDD referred for ECT were randomly assigned to either ECT or rTMS. Repetitive transcranial magnetic stimulation was performed at 90% power of the motor threshold. The stimulation frequency was 10 Hz for either 2 sec (first eight patients) or 6 sec (final 12 patients) for 20 trains. Patients were treated for up to 20 treatment days. Electroconvulsive therapy was performed according to standard protocols. RESULTS: Overall patients responded best to ECT (chi(2) = 3.8, p <.05). Patients with MDD and psychosis responded significantly better to ECT (chi(2) = 9.2, p <. 01), whereas MDD patients without psychosis responded similarly to both treatments (chi(2) = 0.0, ns). The analysis of variance with repeated measures of clinical variables for the whole sample revealed significant treatment effects for both groups; however, interaction between group and treatment was seen only for the Global Assessment of Function and the Sleep assessment. When the psychosis nonpsychosis grouping was considered, patients with psychosis responded dramatically better to ECT in all assessments, whereas those without psychosis responded similarly to both treatments. CONCLUSIONS: Overall ECT was a more potent treatment for patients with MDD, this being particularly evident in patients with MDD and psychosis; however, in patients with MDD without psychosis the effects of rTMS were similar to those of ECT. The results we report are encouraging and support an important role for rTMS in the treatment of severe MDD; however, additional blinded studies are needed to precisely define this role. PMID- 10686267 TI - Transcranial magnetic stimulation (TMS) in controlled treatment studies: are some "sham" forms active? AB - BACKGROUND: Carefully designed controlled studies are essential in further evaluating the therapeutic efficacy of transcranial magnetic stimulation (TMS) in psychiatric disorders. A major methodological concern is the design of the "sham" control for TMS. An ideal sham would produce negligible cortical stimulation in conjunction with a scalp sensation akin to real treatment. Strategies employed so far include alterations in the position of the stimulating coil, but there has been little systematic study of their validity. In this study, we investigated the effects of different coil positions on cortical activation and scalp sensation. METHODS: In nine normal subjects, single TMS pulses were administered at a range of intensities with a "figure eight" coil held in various positions over the left primary motor cortex. Responses were measured as motor-evoked potentials in the right first dorsal interosseus muscle. Scalp sensation to TMS with the coil in various positions over the prefrontal area was also assessed. RESULTS: None of the coil positions studied met the criteria for an ideal sham. Arrangements associated with a higher likelihood of scalp sensation were also more likely to stimulate the cortex. CONCLUSIONS: The choice of a sham for TMS involves a trade-off between effective blinding and truly inactive "stimulation." Further research is needed to develop the best sham condition for a range of applications. PMID- 10686268 TI - A randomized clinical trial of repetitive transcranial magnetic stimulation in the treatment of major depression. AB - BACKGROUND: Multiple groups have reported on the use of repetitive transcranial magnetic stimulation (rTMS) in treatment-resistant major depression. The purpose of this study is to assess the efficacy of rTMS in unmedicated, treatment resistant patients who meet criteria for major depression. METHODS: Depressed subjects, who had failed to respond to a median of four treatment trials, were assigned in a randomized double-blind manner to receive either active (n = 10; 20 2-sec trains of 20 Hz stimulation with 58-sec intervals; delivered at 80% motor threshold with the figure-of-eight coil positioned over the left dorsolateral prefrontal cortex) or sham (n = 10; similar conditions with the coil elevated and angled 45 degrees tangentially to the scalp) rTMS. These sequences were applied during 10 consecutive weekdays. Continuous electroencephalogram sampling and daily motor threshold determinations were also obtained. RESULTS: The group mean 25-item Hamilton Depression Rating Scale (HDRS) score was 37.2 (+/- 2.0 SEM) points. Adjusted mean decreases in HDRS scores were 14.0 (+/- 3.7) and 0.2 (+/- 4.1) points for the active and control groups, respectively (p <.05). One of 10 subjects receiving active treatment demonstrated a robust response (i.e., HDRS decreased from 47 to 7 points); three other patients demonstrated 40-45% decreases in HDRS scores. No patients receiving sham treatment demonstrated partial or full responses. CONCLUSIONS: A 2-week course of active rTMS resulted in statistically significant but clinically modest reductions of depressive symptoms, as compared to sham rTMS in a population characterized by treatment resistance. PMID- 10686269 TI - Ultradian rhythms and temporal coherence in sleep EEG in depressed children and adolescents. AB - BACKGROUND: It has been suggested that a primary ultradian (80-120 minute) rhythm disturbance in EEG underlies sleep abnormalities in adults with depression. The present study evaluated ultradian rhythm disturbances in childhood and adolescent depression. METHODS: Sleep macroarchitecture and temporal coherence in quantitative EEG rhythms were investigated in 50 medication-free outpatients with major depression (25 children and 25 adolescents) and 15 healthy normal controls (5 children and 10 adolescents). RESULTS: Few of the macroarchitectural measures showed significant group effects. In fact, age and sex effects were stronger than disease-dependent components. Temporal coherence of EEG rhythms during sleep did differentiate those with MDD from controls. Both depressed children and adolescents had lower intrahemispheric coherence, whereas interhemispheric was only lower in depressed adolescents in comparison with controls. Gender differences were evident in adolescents, but not children, with MDD with lowest interhemispheric coherence in adolescent girls. CONCLUSIONS: These findings are in keeping with increased risk for depression in females beginning at adolescence and extending throughout adulthood. It was suggested that low temporal coherence in depression reflects a disruption in the fundamental basic rest-activity cycle of arousal and organization in the brain that is strongly influenced by gender. PMID- 10686270 TI - Antidepressant effects of ketamine in depressed patients. AB - BACKGROUND: A growing body of preclinical research suggests that brain glutamate systems may be involved in the pathophysiology of major depression and the mechanism of action of antidepressants. This is the first placebo-controlled, double-blinded trial to assess the treatment effects of a single dose of an N methyl-D-aspartate (NMDA) receptor antagonist in patients with depression. METHODS: Seven subjects with major depression completed 2 test days that involved intravenous treatment with ketamine hydrochloride (.5 mg/kg) or saline solutions under randomized, double-blind conditions. RESULTS: Subjects with depression evidenced significant improvement in depressive symptoms within 72 hours after ketamine but not placebo infusion (i.e., mean 25-item Hamilton Depression Rating Scale scores decreased by 14 +/- SD 10 points vs. 0 +/- 12 points, respectively during active and sham treatment). CONCLUSIONS: These results suggest a potential role for NMDA receptor-modulating drugs in the treatment of depression. PMID- 10686271 TI - Body temperature and mood variations during forced desynchronization in winter depression: a preliminary report. AB - BACKGROUND: It has been suggested that certain abnormalities (e.g., in phase or amplitude) of the circadian pacemaker underlie seasonal affective disorder. METHODS: One male seasonal affective disorder patient (blind to the study design) participated in two 120-hour forced desynchrony experiments and was subjected to six 20-hour days, once during a depressive episode and once after recovery. Core body temperature was continuously measured. During wakefulness, the Adjective Mood Scale was completed at 2-hour intervals. RESULTS: Sleep-wake as well as pacemaker-related variations of mood were found, both when the subject was depressed and when he was euthymic. Compared with recovery, during the depressive episode the circadian temperature minimum and the circadian mood variation showed phase delays of approximately 1 and 2 hours, respectively. CONCLUSIONS: The data of this first seasonal affective disorder patient, participating in forced desynchrony experiments, may indicate a phase delay of the circadian pacemaker during a seasonal affective disorder episode. PMID- 10686272 TI - Glutamine synthetase in experimental meningitis: increased ratio of the subunits 3 and 2 may indicate enhanced activity. AB - Glutamine synthetase (GS) activity is higher in the neocortex but not in the hippocampal formation of rabbit brain during Streptococcus pneumoniae meningitis compared to the respective brain region of uninfected control animals. One dimensional polyacrylamide gel electrophoresis (1D-SDS-PAGE) revealed an apparent molecular mass (M(r)) of 44000 Dalton (Da) for GS from rabbit brain. After two dimensional gel electrophoresis (2D-PAGE), followed by Coomassie-blue staining, GS separated into three distinct spots (S1, S2, S3). One additional spot (S4) occurred on the immunoblot. All four GS spots exhibited the same M(r) (44000 Da), but differed in their isoelectric points. Densitometric evaluation of the two dimensional maps revealed a strong increase of optical density (OD) of S3 in the frontal cortex of infected animals. The calculated OD ratio S3/S2 in the frontal cortex from rabbits with meningitis was 1.75+/-0.68 (mean+/-standard deviation). Compared to controls (0. 85+/-0.39), this value was significantly increased (p=0.0006). In the hippocampal formation, the ratio S3/S2 was nearly unchanged during meningitis. It is suggested that the ratio S3/S2 may indicate a neuroprotective feature of rabbit brain during meningitis since neuronal apoptosis occurs only in the dentate gyrus and not in the frontal cortex. PMID- 10686273 TI - Performance of the enhanced Abbott AxSYM cardiac troponin I reagent in patients with heterophilic antibodies. AB - The presence of heterophilic antibodies in the serum of a small subpopulation of individuals continues to cause false results for modern-day immunoassays. In order to determine the frequency of heterophilic antibody (HA)-related false positives within our population of positive cardiac troponin I (cTnI) patients, we assayed 200 samples using the original in-house cTnI assay (Abbott AxSYM) and the Bayer ACS:180 cTnI, which we had previously observed to be more effective at blocking HA interference. Four samples were identified as false positives based on discordant results between the two assays, as well as the correction of the false positives by treatment of the samples with heterophilic antibody blocking reagent (HBR). An 'enhanced' version of the AxSYM cTnI reagent was designed to greatly reduce or eliminate HA interference, and has now replaced the original reagents. The present study shows that the enhanced reagent significantly reduced or eliminated much of the HA interference. Comparative studies between the enhanced cTnI reagent and the original Abbott AxSYM cTnI reagent showed excellent correlation and equivalent diagnostic concordance, when HA samples were excluded from the analysis. PMID- 10686274 TI - Proteolytic activation of purified human procarboxypeptidase U. AB - Carboxypeptidase U (CPU, EC 3.4.17.20) is a recently described basic carboxypeptidase which circulates in plasma as an enzymatically inactive precursor procarboxypeptidase U (proCPU), also known as plasma carboxypeptidase B precursor or thrombin activatable fibrinolysis inhibitor (TAFI). The activation of the zymogen proceeds through a proteolytic cleavage at Arg-92. The active form - CPU - is able to retard the initial phase of fibrinolysis by cleaving C terminal lysine residues exposed on fibrin partially degraded by the action of plasmin. These C-terminal lysine residues are essential for the high affinity binding of plasminogen to fibrin and the subsequent activation to plasmin. In this report, the activation of purified human proCPU was studied using trypsin and some key proteases of the coagulation and fibrinolytic cascade, i.e., kallikrein, plasmin and thrombin. The most efficient activation is obtained in the presence of thrombin in complex with thrombomodulin. After in vitro activation, CPU is unstable at 37 degrees C (T(1/2)=15 min). Its stability can be improved dramatically using lower temperatures. PMID- 10686275 TI - Improved assay for fecal calprotectin. AB - Fecal calprotectin is a marker of inflammatory and neoplastic disease in the lower gastrointestinal tract. A new fecal sample preparation procedure for the measurement of calprotectin has been developed, with higher calprotectin yield and lower contamination risk. Changes in the new method compared to the original [Roseth AG, Fagerhol MK, Aadland E, Schonsby H. Assessment of the neutrophil dominating protein calprotectin in feces. A methodologic study. Scand J Gastroenterol 1992;27(9):793-798] are smaller sample size, higher dilution of the sample, presence of dissociating agents in the extraction solution and procedure performed in closed disposable tubes. The extraction yield was 78% (41-100%) of total calprotectin, giving an overall five-fold increase compared to the original method. Samples with high calprotectin values were increased to a slightly higher degree, than low calprotectin samples, thus improving the separation between high and low calprotectin levels. Median calprotectin level in healthy subjects was 26 microg/g. Pathological samples with pancolitis showed levels up to 30000 microg/g. The mean C.V. (coefficient of variation) in blended feces was lower than that of unblended, suggesting uneven distribution of calprotectin. However, no significant difference between spot measurements was found when five samples from each of 47 stools were measured. Thus measurements of calprotectin in fecal samples were accurate and reproducible. No interference with foods or relevant oral pharmaceuticals or nutraceuticals was found. PMID- 10686276 TI - Evaluation of intrinsic and routine quality of serum total magnesium measurement. AB - We investigated the intrinsic (as delivered by the manufacturer) and routine quality of four systems for measurement of serum total magnesium (t-Mg(2+)) by method comparison with an ion chromatography reference method. The results of the study were interpreted on the basis of analytical quality specifications derived from the biological variation of t-Mg(2+), expanded by the analytical uncertainty of the reference measurements. This resulted in limits for systematic error of 2.1% and for total error of 4.3%. The study demonstrated that those limits were challenging for all routine systems. Most of them met the total error criterium just borderline and one showed a considerable systematic error (-5.2%). Concerning the measurement quality in the routine laboratories, the study showed that many were unable to preserve the intrinsic quality of the respective manufacturer. Consequently, loss of system performance in the routine laboratory mostly led to violation of the analytical specifications. Most strikingly, the study revealed enormous quality differences between routine laboratories. This indicates that, still, many routine laboratories do not make adequate use of currently available internal and external quality control tools. Moreover, some laboratories considerably expanded the high end of the reference interval, thereby reducing the diagnostic potential of t-Mg(2+). PMID- 10686277 TI - Prebeta1-high-density lipoprotein (prebeta1-HDL) concentration can change with low-density lipoprotein-cholesterol (LDL-C) concentration independent of cholesteryl ester transfer protein (CETP). AB - To clarify whether prebeta1-high-density lipoprotein (prebeta1-HDL) concentration changes with low-density lipoprotein-cholesterol (LDL-C) concentration independent of cholesteryl ester transfer protein (CETP), we determined prebeta1 HDL concentration by native two-dimensional gel electrophoresis in 58 subjects with normal triglyceride and HDL-cholesterol concentrations. We also measured LDL C and CETP concentrations. In 17 subjects, a second blood sample was taken 1-6 months after the first. We found that prebeta1-HDL concentration was positively correlated with LDL-C concentration (r=0.529, P<0.0001) and with CETP mass (r=0.398, P<0.01). In 17 patients, Deltaprebeta1-HDL was positively correlated with DeltaLDL-C (r=0.635, P<0.01), but not with DeltaCETP mass (r=0.275). In conclusion, prebeta1-HDL concentration changes with LDL-C concentration independent of CETP. These results suggest that prebeta1-HDL concentration may reflect the balance between several regulatory factors, including LDL-C and CETP concentrations. PMID- 10686278 TI - Plasma levels of coenzyme Q(10), vitamin E and lipids in uremic patients on conservative therapy and hemodialysis treatment: some possible biochemical and clinical implications. AB - Coenzyme Q(10) (CoQ(10)), vitamin E, total cholesterol, HDL-cholesterol (HDLC) and triglycerides were measured in the plasma of 62 patients with kidney failure, 46 under hemodialysis treatment and 16 under conservative therapy, and 95 controls. The sum of LDL-cholesterol (LDL-C) and VLDL-cholesterol (VLDL-C) was also calculated for each patient. The ratio CoQ(10)/LDL-C+VLDL-C in both conservative therapy and hemodialysis populations was significantly lower (P<0.001) compared with normal controls and remained unchanged after the dialysis treatment. On the contrary the ratio vitamin E/LDL-C+VLDL-C was normal but decreased significantly (P<0.02) after each dialysis. Since coenzyme Q is the main inhibitor of the prooxidant action of vitamin E, it was hypothesized that its decrease in both the populations examined could make the lipoproteins of these patients more vulnerable to a peroxidative attack. PMID- 10686279 TI - Stereodifferentiation of 3-hydroxyisobutyric- and 3-aminoisobutyric acid in human urine by enantioselective multidimensional capillary gas chromatography-mass spectrometry. AB - The chiral metabolites 3-hydroxyisobutyric acid (HIBA) and 3-aminoisobutyric acid (AIBA) are intermediates in the pathways of L-valine and thymine and play an important role in the diagnosis of the very rare inherited metabolic diseases 3 hydroxyisobutyric aciduria (McKusick 236975) and methylmalonic semialdehyde dehydrogenase deficiency (McKusick 603178-MSDD). Until now only a few approaches have been made in enantioselective analysis of HIBA and AIBA and for that reason very little information is available on enantiomeric ratios of these metabolites in man. This paper reports on the simultaneous stereodifferentiation of HIBA and AIBA in human urine as corresponding N(O)-methoxycarbonyl methyl esters by derivatization with methyl chloroformate (MCF) using enantioselective multidimensional gas chromatography-mass spectrometry (enantio-MDGC/MS) with heptakis-(2, 3-di-O-methyl-6-O-tert.-butyl-dimethylsilyl)-beta-cyclodextrin as the chiral stationary phase. During this investigation urine samples from different patients and healthy controls were analyzed in order to reveal characteristic enantiomeric patterns of these metabolites. A trend of dominating R-HIBA excretion in the control urine samples investigated was observed. An excretion of more than 80% S-HIBA was found in the urines of two patients with ketonemic vomiting. There are some clues indicating a possible renal reabsorbtion of S-HIBA similar to those of S-AIBA. Furthermore, there was a significant finding with regard to the enantiomeric distribution of AIBA in a patient with MSDD - a markedly increased excretion of the S-enantiomer in contrast to the other samples. Using the enantiomeric ratios of AIBA, a previously investigated case of benign methylmalonic aciduria (bMMA) could be excluded from the diagnosis of MSDD. PMID- 10686280 TI - Determination of serum procarboxypeptidase A concentrations in healthy human adults. AB - Clostripain (EC 3.4.22.8) has been shown to be superior to trypsin as a means of activating serum procarboxypeptidase A. With this activation and a previously described assay for carboxypeptidase A it has been possible to determine the concentration of procarboxypeptidase A in human serum. In order to establish a baseline in the healthy adult a survey of the procarboxypeptidase A content of the serum of 66 blood donors was carried out. An average value of 9.7 microg/l was found for the proenzyme. This is in sharp contrast to a mean of 0.34 microg/l for the free enzyme present in serum that was not treated with clostripain [1]. PMID- 10686281 TI - The effect of reaction temperature for nephelometric assays for rheumatoid factor. AB - Even using the same assay parameter, reagent and calibrator (N-latex RF kit II), the results of the assay for serum rheumatoid factors (RFs) with the Behring Nephelometer Analyzer (BNA) were higher than those with the Behring Nephelometer II Analyzer (BNII) ([BNII]=0.76 [BNA]-5.7 kIU/l, r=0.997, Sy/x=60.73, n=99). The mean bias (BNA minus BNII)+/-S.D. was 52.7+/-85.5 using the Bland and Altman plot method, and the bias was not constant. The only difference in the assay condition with the two methods was the reaction temperature with the BNA being performed at room temperature (25+/-1 degrees C) and the BNII being performed at 37 degrees C. The ratio of the results with the BNII to the BNA (BNII/BNA) ranged from 0.23 to 1.18. A significant difference was observed in the BNII/BNA ratio in patients with high levels of C-reactive protein (CRP) over 2.0 mg/l (mean BNII/BNA ratio; 0.78) in comparison to patients with normal CRP levels under 2.0 mg/l (mean BNII/BNA ratio; 0.65) (P<0.01). The RF concentrations with the BNA were reduced by addition of urea, which has been used as a mild protein-denaturing agent, and there was a significant correlation between the values calculated as (1-value treated with urea/original value without urea)x100 and the BNII/BNA ratio (r=0.652, P<0.01). These data suggested that the bias between the RF values obtained by the BNA and BNII might be caused by the variation in the reactivity of autoantibodies, which might be decreased in some inflammatory diseases. PMID- 10686282 TI - Comparison of the percent free prostate-specific antigen levels in the serum of healthy men and in men with recurrent prostate cancer after radical prostatectomy. AB - The percentage of free PSA in serum is currently used to better discriminate between patients with prostate cancer and patients with benign prostatic hyperplasia, in prostate cancer screening programs. We measured using non competitive immunological techniques, the total PSA and free PSA in post-surgical serum of prostate cancer patients who underwent radical prostatectomy and then relapsed. We compared these data with those of a group of 40 age-matched men with no evidence of prostatic disease. Although in general, patients with prostate cancer had lower percentage of free PSA in serum in comparison to the controls, a subset of these patients (approximately 20%) had percent free PSA significantly higher than the levels considered as exclusive of prostate cancer in screening programs. We also found that percent free PSA does not correlate significantly with most of the standard clinical or pathological indicators of prostate cancer aggressiveness. Only a weak negative association with Gleason Score was observed. The percent free PSA in serum of relapsing prostate cancer patients varies within a relatively wide range and does not correlate significantly with indicators of cancer aggressiveness. The use of percent free PSA for excluding prostate cancer in screening programs must be approached with caution until the mechanism of low percent free PSA in the majority but not all prostate cancer patients is elucidated. PMID- 10686283 TI - Selection of an optimal combination of decision criteria for the internal quality control in an automatic analyzer. AB - After some years using an automatic analyser model Hitachi 705, we faced the task of selecting a reasonable combination of statistical procedures for internal quality control in order to use it in a computer program, by means of which, one could apply all the decision criteria in a rapid and automatic manner. As a result of the first 3 years of work, an algorithm was adopted that includes the multirule procedure originally proposed by Westgard, but with three main differences: (1) All the control rules are always applied; (2) the 4(1S) and 10x control rules are used as criteria for warning or caution but not for run rejection; and (3) the control limits are recalculated daily using all the accumulated control results (just the accepted values). The 2(2S), 4(1S) and 10x rules are applied first across materials (considering consecutive observations in different control materials), and then within materials (for consecutive observations on the same control material). The program not only has permitted us to improve our capacity for error detection, but it has also permitted the reduction of run rejections making an important decrease in costs possible and a significant improvement in quality. PMID- 10686284 TI - Mild renal dysfunction is sufficient to induce erythropoietin deficiency in patients with unexplained anaemia. AB - Current guidelines suggest that anaemia due to erythropoietin deficiency almost exclusively occurs with creatinine concentrations of at least 177 micromol/l or above. The aim of this prospective case control pilot study was to evaluate whether borderline renal function or mild renal dysfunction with creatinine concentrations well below 177 micromol/l is sufficient to induce inadequate erythropoietin secretion. Patients referred for work-up of otherwise unexplained anaemia with mildly abnormal creatinine concentrations (104-129 micromol/l; study group: eight patients) and patients referred for work-up or therapy of other diseases who also presented with anaemia but normal creatinine levels (<100 micromol/l; control group: nine patients matched for gender, age and degree of anaemia) were included. All but two patients in the control group had bone marrow biopsies to exclude other pathologies. Mild renal dysfunction (as evidenced by creatinine concentrations between 100 and 140 micromol/l, median concentration 112 micromol/l) was found to be sufficient to induce inadequate erythropoietin secretion. The physiologic hemoglobin-dependent erythropoietin regulation demonstrated in the control group was abolished in the study group. Patients with mild renal dysfunction and unexplained anaemia should be investigated for erythropoietin concentration. If the erythropoietin concentration is found to be inadequate for the degree of anaemia, substitution therapy should be considered. PMID- 10686285 TI - Analysis of leukotrienes in cerebrospinal fluid of a reference population and patients with inborn errors of metabolism: further evidence for a pathognomonic profile in LTC(4)-synthesis deficiency. AB - Cysteinyl leukotrienes (LTC(4), LTD(4), LTE(4)) are potent lipid mediators derived from arachidonate in the 5-lipoxygenase pathway. Recently, the first inborn error of leukotriene synthesis, LTC(4)-synthesis deficiency, has been identified in association with a fatal developmental syndrome. The absence of leukotrienes in cerebrospinal fluid was one of the most striking biochemical findings in this disorder. We analysed leukotrienes in cerebrospinal fluid of patients with a broad spectrum of other well-defined inborn errors of metabolism, including glutathione synthetase deficiency (n=2), Zellweger syndrome (n=3), mitochondrial disorders (n=8), fatty acid oxidation defects (n=7), organic acidurias (n=7), neurotransmitter defects (n=5) and patients with non-specific neurological symptoms, as a reference population (n=120). The concentrations of leukotrienes were not related to age. Representative percentiles were calculated as reference intervals of each leukotriene. In all patients with an inborn error of metabolism concentration of cysteinyl leukotrienes and LTB(4) did not differ from the reference group. Our results indicate that absence of cysteinyl leukotrienes (<5 pg/ml) in association with normal or increased LTB(4) (50.0-67.3 pg/ml) is pathognomonic for LTC(4)-synthesis deficiency. The unique profile of leukotrienes in cerebrospinal fluid in this new disorder is primarily related to the defect and represents a new diagnostic approach. PMID- 10686286 TI - Reference values for selected trace elements in serum of term newborns from the urban area of Rome. AB - Reference values for Al, Cd, Co, Cu, Li, Mn, Mo, Ni, Rb, Se and Zn, and indicative intervals for Sb are proposed in serum from cord blood of 143 term newborns of the urban area of Rome. On the basis of the eligibility criteria adopted, only babies with gestational age > 37 weeks and body weight at the delivery > 2500 g, i.e., "normal" term infants, were included in this study. With the exception of Cd, Li, Ni and Sb, experimental data for each of the other analytes were found to approach a normal distribution. The estimated references values (in ng/ml) were the following: Al, 1. 12-6.79; Cd, 0.10-0.52; Co, 0.20 0.43; Cu, 140-691; Li, 0.31-2.23; Mn, 0.79-3.26; Mo, 0.36-1.56; Ni, 0.20-3.15; Rb, 196-1302; Sb, 0. 10-1.48 (indicative range); Se, 20.2-69.7; and Zn, 318-1405. For several elements, the information available in the relevant literature does not allow adequate comparisons to be performed. This was actually possible only for Cu, Se and Zn. The correlations between the weights at birth (BW), gestational ages (GA) and elemental concentrations were elucidated. As expected, significant positive correlations were found for Cu and Se with GA and BW, respectively. Strong mutual associations were observed for several other elements, but their interpretation is still debatable. PMID- 10686287 TI - Correcting serum fructosamine concentration for total protein or albumin concentration is not appropriate during Asian pregnancy. PMID- 10686288 TI - Serum dipeptidylpeptidase activity and tissue polypeptide specific antigen in patients with advanced cirrhosis: preliminary results. PMID- 10686290 TI - Alpha 1-adrenoceptors: function and phosphorylation. AB - This review focuses on alpha(1)-adrenoceptor phosphorylation and function. Most of what is currently known is based on studies on the hamster alpha(1B) adrenoceptor. It is known that agonist stimulation leads to homologous desensitization of these receptors and current evidence indicates that such decrease in receptor activity is associated with receptor phosphorylation. Such receptor phosphorylation seems to involve G protein-receptor kinases and the receptor phosphorylation sites have been located in the carboxyl tail (Ser(404), Ser(408), and Ser(410)). There is also evidence showing that in addition to desensitization, receptor phosphorylation is associated with internalization and roles of beta-arrestins have been observed. Direct activation of protein kinase C leads to receptor desensitization/internalization associated with phosphorylation; the protein-kinase-C-catalyzed receptor phosphorylation sites have been also located in the carboxyl tail (Ser(394) and Ser(400)). Activation of G(q)-coupled receptors, such as the endothelin ET(A) receptor induces alpha(1B)-adrenoceptor phosphorylation and desensitization. Such effect involves protein kinase C and a yet unidentified tyrosine kinase. Activation of G(i) coupled receptors, such as the lysophosphatidic acid receptor, also induces alpha(1B)-adrenoceptor phosphorylation and desensitization. These effects involve protein kinase C and phosphatidyl inositol 3-kinase. Interestingly, activation of epidermal growth factor receptors also induces alpha(1B)-adrenoceptor phosphorylation and desensitization involving protein kinase C and phosphatidyl inositol 3-kinase. A pivotal role of these kinases in heterologous desensitization is evidenced. PMID- 10686291 TI - Proteolysis and phosphorylation-mediated regulation of thrombin receptor activity in in situ endothelial cells. AB - The regulatory mechanism of thrombin receptor responsiveness in in situ endothelial cells was investigated by evaluating elevations of cytosolic Ca(2+) concentration ([Ca(2+)](i)) in fura-2-loaded porcine aortic valvular strips. Once stimulated with thrombin, endothelial cells did not respond to the second thrombin stimulation within 90 min. However, applying thrombin receptor activating peptide (TRAP7) at 15 min after the thrombin stimulation caused [Ca(2+)](i) elevation, which was smaller than that seen without preceding stimulation. After 90 min, response to TRAP7 recovered to the control level. When stimulated with TRAP7, the subsequent responses to thrombin and TRAP7 were attenuated at 15 min, and fully recovered after 90 min. Staurosporine partially prevented the TRAP7-induced desensitization. The recovery of responsiveness was inhibited completely by calyculin-A and partially by okadaic acid. Proteolysis and phosphorylation thus play an important role in thrombin receptor desensitization in in situ endothelial cells. Both cleaved and uncleaved receptors were desensitized through phosphorylation in part by staurosporine sensitive kinase, and restored the responsiveness through dephosphorylation by type 1 phosphatase. The mechanism of regulation of thrombin receptor activity in in situ endothelial cells differed from those reported in cultured endothelial cells. We suggest that the cell-specific regulatory mechanism may be altered by culture conditions. PMID- 10686292 TI - Prostanoids regulate proliferation of vascular smooth muscle cells induced by arginine vasopressin. AB - The aim of the present study was to investigate the effect of arginine [Arg(8)]vasopressin (vasopressin) on proliferation of vascular smooth muscle cells and the mechanisms underlying the action of vasopressin. To clarify these issues, we used two different types of vascular smooth muscle cells, cultured adult rat aortic smooth muscle cells and A10 cells, a cell line derived from fetal rat aorta. Vasopressin (10(-8) to 10(-6) M) significantly stimulated the proliferation of rat aortic smooth muscle cells in a dose-dependent manner. In contrast, vasopressin significantly inhibited the proliferation of A10 cells. This inhibition was abolished when A10 cells were treated with indomethacin. Vasopressin stimulated the production of prostanoids several-fold in A10 cells but not in rat aortic smooth muscle cells. These effects were completely blocked by the vasopressin V(1) receptor antagonist, 1-?1-[4-(3-acetylamino propoxy)benzoyl]4-piperidyl?-3, 4-dihydro-2(1H)-quinolinone (OPC21268), but not by the vasopressin V(2) receptor antagonist, (+/-)-5-dimethylamino-1-[4-(2 methylbenzoylamino)benzol]-2, 3,4,5-tetrahydro-1H-benzazepine hydrochloride (OPC31260). These results indicate that vasopressin has diverse effect on proliferation of vascular smooth muscle cells through the vasopressin V(1) receptor, depending on the production of growth regulatory prostanoids. PMID- 10686293 TI - Tyrosine kinase inhibitors and Ca2+ signaling: direct interactions with fura-2. AB - Selective inhibitors were used to study the role of tyrosine kinases in alpha(1A) adrenoceptor-mediated responses in transfected PC12 cells. Ca(2+) responses to noradrenaline were measured using fura-2, and the effects of genistein, tyrphostin A25, and herbimycin A were examined. Neither genistein nor herbimycin A pretreatment altered noradrenaline-induced Ca(2+) responses, although tyrphostin A25 pretreatment caused some reduction. However, acute addition of genistein quickly reversed the apparent noradrenaline response, apparently, through a direct interaction with cytoplasmic fura-2. Both genistein and tyrphostin A25, at concentrations similar to those used to inhibit tyrosine kinases, markedly reduced fluorescence of fura-2 excited by both 340 and 380 nm, and genistein also reduced the 340/380 ratio. Tyrosine kinase inhibitors did not alter noradrenaline stimulated inositol phosphate formation in alpha(1A)-PC12 cells. These results suggest that tyrosine kinases are not involved in second messenger responses to alpha(1A)-adrenoceptors, but that tyrosine kinase inhibitors can interact directly with fura-2. PMID- 10686294 TI - Species selectivity of a small molecule antagonist for the CCR1 chemokine receptor. AB - The species specificity of a small molecule antagonist for the human CCR1 chemokine receptor, 2-2-diphenyl-5-(4-chlorophenyl)piperidin-1-yl)valeronitrile (CCR1 antagonist 1), has been examined using cloned CCR1 receptors from various species. The compound was able to bind to rabbit, marmoset, and human CCR1, and was able to block the functional activation of these receptors. However, it failed to significantly displace radiolabeled macrophage inflammatory protein 1alpha (MIP-1alpha) binding to mouse CCR1 at concentrations up to 10 microM. These data suggested that the antagonist binding site is well-conserved in rabbit, marmoset and human CCR1, but not in mouse CCR1. The functional selectivity and mechanism of action for CCR1 antagonist 1 were further characterized. CCR1 antagonist 1 blocked the increase in intracellular Ca(2+) stimulated by CCR1 agonists, but had no effect on N-formyl-Met-Leu-Phe (FMLP), monocyte chemotactic protein-1 (MCP-1) and stromal-derived factor 1alpha (SDF1alpha)-induced Ca(2+) mobilization, demonstrating functional selectivity for CCR1. Since CCR1 antagonist 1 is a functional antagonist of marmoset and rabbit CCR1 receptors, it should be possible to test its efficacy in animal models of disease. PMID- 10686295 TI - Cromakalim-induced membrane current in guinea-pig tracheal smooth muscle cells. AB - The characteristics of the cromakalim-induced membrane current were examined in single tracheal myocytes of the guinea-pig under voltage-clamp conditions. When K(+) concentrations in the pipette and bathing solutions were approximately 140 mM, cromakalim activated a membrane current (I(crom)) which was inward at -60 mV and reversed at -2 mV. I(crom) was blocked by 10 microM glibenclamide and potentiated when the ATP concentration in the pipette solution was decreased. The K(d) and Hill coefficient of glibenclamide for I(crom) block were 200 nM and 1.05, respectively. Application of the tyrosine kinase inhibitors, genistein and alpha-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamamid (ST638), reduced I(crom) in a concentration-dependent manner. Daidzein, which does not inhibit tyrosine kinase, was about 10 times less effective than genistein. Herbimycin A had no effect on I(crom). Internal application of these inhibitors from the pipette did not affect I(crom). In conclusion, cromakalim is a potent activator of the ATP-sensitive K(+) channel (K(ATP) channel) in guinea-pig tracheal myocytes. The inhibition of I(crom) by genistein and ST638 may be due to the direct block of the channel from outside. PMID- 10686296 TI - Dopamine transporter antagonists block phorbol ester-induced dopamine release and dopamine transporter phosphorylation in striatal synaptosomes. AB - We have reported that inhibition of protein kinase C blocks the Ca(2+) independent reverse transport of dopamine mediated by amphetamine. In this study we investigated whether activation of protein kinase C by 12-O-tetradecanoyl phorbol-13-acetate (TPA) would mediate dopamine release through the plasmalemmal dopamine transporter. TPA, at 250 nM, increased the release of dopamine from rat striatal slices and synaptosomes while the inactive phorbol ester, 4alpha phorbol, was ineffective. The TPA-mediated dopamine release was independent of extracellular calcium and was blocked by a selective protein kinase C inhibitor, Ro31-8220. The dopamine transporter antagonists, cocaine and GBR 12935 blocked the TPA-mediated dopamine release. In addition, cocaine blocked TPA-mediated phosphorylation of the plasmalemmal dopamine transporter. These results suggest that activation of protein kinase C results in reverse transport of dopamine through the plasmalemmal dopamine transporter and the phosphorylated substrate could be the dopamine transporter. PMID- 10686297 TI - Cyclooxygenase-independent inhibition of smooth muscle cell mitogenesis by ibuprofen. AB - The aryl-propionic acid derivative, ketoprofen, has been shown to inhibit fibroblast growth by a cylooxygenase-dependent mechanism [Sanchez, T., Moreno, J.J., 1999. S(+) enantiomer inhibits prostaglandin production and cell growth in 3T6 fibroblast cultures. Eur. J. Pharmacol. 370, 63-67]. The present study demonstrates that ibuprofen, another aryl-propionic acid derivative, inhibited platelet-derived growth factor-BB (20 ng/ml)-induced mitogenesis of cultured bovine coronary artery smooth muscle cells in a stereo-independent manner. In addition, pretreatment of the cells with indomethacin (3 microM) did not affect the inhibitory effects of ibuprofen enantiomers on smooth muscle cell mitogenesis. Thus, aryl-propionic acid-type cyclooxygenase inhibitors can inhibit cell proliferation by both, cyclooxygenase-dependent and -independent ways. PMID- 10686298 TI - Differential role of nitric oxide pathway and heat shock protein in preconditioning and lipopolysaccharide-induced brain ischemic tolerance. AB - The purposes of this study were to investigate the role of nitric oxide (NO), nitric oxide synthase (NOS), and 70 kDa heat shock protein in brain ischemic tolerance induced by ischemic preconditioning and lipopolysaccharide. Focal cerebral ischemia was induced in rats by intraluminal middle cerebral artery occlusion. Infarct volume was significantly reduced (1) in rats subjected to 3 min ischemia 72 h prior to 60 min ischemia; (2) in rats administered lipopolysaccharide (0.5 mg/kg; i.p.) 72 h prior to 60 min ischemia compared with controls. The beneficial effect of ischemic preconditioning was unchanged despite prior administration of nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor. Conversely, the protective effect of lipopolysaccharide was nullified by L-NAME. Using immunohistochemical techniques, we observed that (1) ischemic preconditioning but not lipopolysaccharide induces the expression of 70 kDa heat shock protein in cerebral cortex and (2) lipopolysaccharide induces early increased expression of endothelial NOS in cerebral blood vessels. The results suggest that (1) endothelium-derived NO plays a role of a trigger in the brain tolerance induced by lipopolysaccharide, and (2) 70 kDa heat shock protein is involved in the protection afforded by ischemic preconditioning but not by lipopolysaccharide. PMID- 10686299 TI - Neuroprotection by LY341122, a novel inhibitor of lipid peroxidation, against focal ischemic brain damage in rats. AB - LY341122 (2-(3, 5-di-t-butyl-4-hydroxyphenyl)-4-(2-(4-methylethylaminomethyl-ph enylox y)ethyl)oxazole) is a potent inhibitor of lipid peroxidation which has been shown to protect against global ischemia and traumatic brain injury in rats. The purpose of this study was to examine the effect of LY341122 on ischemic injury in a highly reproducible model of focal cerebral ischemia in rats. Male Sprague-Dawley rats were anesthetized with halothane and subjected to 120 min of temporary middle cerebral artery occlusion by retrograde insertion of an intraluminal nylon suture coated with poly-L-lysine. The drug (LY341122, n=19) or vehicle (phosphate-buffered saline (PBS), n=10) was administered i.v. (as a 5 or 10 mg/kg bolus followed by a 5 or 10 mg/kg/h infusion for 20 h, respectively, starting 1 or 2 h after the onset of middle cerebral artery occlusion). Neurological status was evaluated during middle cerebral artery occlusion (60 min) and daily for 3 days thereafter. Three days after ischemia, brains were perfusion-fixed and infarct volumes and brain edema were determined. LY341122 significantly improved the neurological score compared to vehicle at 24, 48 and 72 h after middle cerebral artery occlusion. Treatment with LY341122 significantly reduced total infarct volume in all treated groups compared to vehicle rats. Cortical infarct volume was significantly reduced by LY341122 treatment in the 10 mg/kg (1 h) and LY341122 10 mg/kg (2 h) groups compared to vehicle rats (14.7+/-9.5 vs. 106.8+/-20.9 mm(3), and 36.9+/-20.1 vs. 106. 8+/ 20.9 mm(3), respectively (mean+/-S.E.M.)). Striatal infarct volume was also significantly reduced by treatment with LY341122 in the 10 mg/kg (1 h) group compared to vehicle (23.7+/-3.4 vs. 68. 2+/-6.7 mm(3)). These results demonstrate the neuroprotective efficacy of LY341122 in focal cerebral ischemia. PMID- 10686300 TI - Endogenous corticotropin-releasing hormone inhibits conditioned-fear-induced vagal activation in the rat. AB - The role of the endogenous corticotropin-releasing hormone (CRH) system in the regulation of heart rate, PQ interval (a measure of vagal activity), gross activity and release of adrenocorticotropic hormone (ACTH), noradrenaline and adrenaline into the blood during conditioned fear was studied in freely moving rats. Intracerebroventricular (i.c.v.) infusion of alpha-helical CRH-(9-41) (10 microgram/3 microliter), a non-selective CRH receptor antagonist, under resting conditions had no significant effect on gross activity, heart rate and PQ interval, indicating that alpha-helical CRH at this dose was devoid of agonist effects. Conditioned fear was induced by 10 min forced exposure to a cage in which the rat had experienced footshocks (5x0.5 mAx3 s) 1 day before. Conditioned fear rats showed freezing behaviour, associated with an increase in heart rate, PQ interval, noradrenaline and adrenaline, indicating that the conditioned-fear induced cardiac effects were the result of coactivation of the sympathetic and parasympathetic nervous system. The i.c.v. pre-treatment of rats with alpha helical CRH significantly reduced the conditioned-fear-induced tachycardiac and ACTH response, and enhanced the increase in PQ interval, without affecting the noradrenaline and adrenaline response. These results suggest that endogenous CRH reduces the vagal response to conditioned-fear stress in rats. To test this, rats were pre-treated with atropine methyl nitrate (0.3 mg/kg, subcutaneously; s.c.), a peripherally acting cholinergic receptor antagonist. This resulted in a complete blockade of the alpha-helical CRH-induced decrease in heart rate response and increase in PQ interval. From these findings, it is concluded that endogenous CRH in the brain inhibits vagal outflow induced by emotional stress. PMID- 10686301 TI - Tachykinin-mediated effect of nociceptin in the rat urinary bladder in vivo. AB - The application of nociceptin (5-50 nmol/rat) onto the serosa in the urinary bladder of urethane-anaesthetized rats, with the intravesical volume kept below threshold for activation of the micturition reflex, induced a low amplitude tonic contraction (local, i.e., resistant to ganglionectomy) with high amplitude phasic contractions (reflex, i.e., abolished by ganglionectomy) superimposed. The pharmacology of the local contraction was studied in animals with acute bilateral ablation in the pelvic ganglia: the combined administration of tachykinin NK(1) (S)1-?2-[3-(3, 4-dichlorophenyl)-1-(3-isopropoxyphenyl-acetyl)-piperidin-3-yl]eth yl?-4-phenyl-1-azoniabicyclo[2.2.2.]octane chloride (SR 140333) and NK(2) c?[(beta-D-GlcNAc)Asn-Asp-Trp-Phe-Dpr-Leu]c(2beta-5beta++ +)? (MEN 11420) receptor antagonists (given at doses of 1+0.1 micromol/kg, intravenous (i.v.), respectively) abolished the local bladder contraction induced by topical nociceptin (50 nmol/rat). These results indicate that the topical application of nociceptin onto the bladder evokes a tachykinin-mediated contraction. PMID- 10686302 TI - Y-27632 potentiates relaxant effects of beta 2-adrenoceptor agonists in bovine tracheal smooth muscle. AB - We examined how (+)-(R)-trans-4-(1-aminoethyl)-N-(4 pyridyl)cyclohexanecarboxamide (Y-27632), an inhibitor of Rho-associated coiled coil-forming protein kinase (ROCK I) and Rho kinase (ROCK II), affects the relaxant responses to beta(2)-adrenoceptor agonists in bovine tracheal smooth muscle preparations precontracted with methacholine. Y-27632 (0.3-30 microM) caused a concentration-dependent attenuation of precontraction with methacholine (0.3-3 microM). Pretreatment with Y-27632 (1 microM) significantly (P<0.05) augmented salbutamol (0.3-100 nM) and terbutaline (0.3 nM-1 microM)-induced relaxations. These results suggest that the ROCK inhibitor could become a new type bronchodilator and its combination with beta(2)-adrenoceptor agonists may become a novel strategy for the long-term treatment of asthma. PMID- 10686303 TI - Paracelsus to parascience: the environmental cancer distraction. AB - Entering a new millennium seems a good time to challenge some old ideas, which in our view are implausible, have little supportive evidence, and might best be left behind. In this essay, we summarize a decade of work, raising four issues that involve toxicology, nutrition, public health, and government regulatory policy. (a) Paracelsus or parascience: the dose (trace) makes the poison. Half of all chemicals, whether natural or synthetic, are positive in high-dose rodent cancer tests. These results are unlikely to be relevant at the low doses of human exposure. (b) Even Rachel Carson was made of chemicals: natural vs. synthetic chemicals. Human exposure to naturally occurring rodent carcinogens is ubiquitous, and dwarfs the general public's exposure to synthetic rodent carcinogens. (c) Errors of omission: micronutrient inadequacy is genotoxic. The major causes of cancer (other than smoking) do not involve exogenous carcinogenic chemicals: dietary imbalances, hormonal factors, infection and inflammation, and genetic factors. Insufficiency of many micronutrients, which appears to mimic radiation, is a preventable source of DNA damage. (d) Damage by distraction: regulating low hypothetical risks. Putting huge amounts of money into minuscule hypothetical risks damages public health by diverting resources and distracting the public from major risks. PMID- 10686304 TI - How we should deal with unavoidable exposure of man to environmental mutagens: cooked food mutagen discovery, facts and lessons for cancer prevention. PMID- 10686305 TI - Formation of DNA adducts and induction of mutagenic effects in rats following 4 weeks inhalation exposure to ethylene oxide as a basis for cancer risk assessment. AB - Ethylene oxide (EO) is mutagenic in various in vitro and in vivo test systems and carcinogenic in rodents. EO forms different adducts upon reaction with DNA, N7-(2 hydroxyethyl)guanine (N7-HEG) being the main adduct. The major objectives of this study were: (a) to determine the formation and persistence of N7-HEG adducts in liver DNA of adult male rats exposed to 0, 50, 100 and 200 ppm by inhalation (4 weeks, 5 days/week, 6 h/day) and (b) to assess dose-response relationships for Hprt gene mutations and various types of chromosomal changes in splenic lymphocytes.N7-HEG adducts were measured 5, 21, 35 and 49 days after cessation of exposure. By extrapolation, the mean concentrations of N7-HEG immediately after cessation of exposure ('day 0') to 50, 100 and 200 ppm were calculated as 310, 558 and 1202 adducts/10(8) nucleotides, respectively, while the mean concentration in control rats was 2.6 adducts/10(8) nucleotides. At 49 days, N7 HEG values had returned close to background levels. The mean levels of N-(2 hydroxyethylvaline) adducts in haemoglobin were also determined and amounted 61.7, 114 and 247 nmol/g globin, respectively. Statistically significant linear relationships were found between mean N7-HEG levels ('day 0') and Hprt mutant frequencies at expression times 21/22 and 49/50 days and between mean N7-HEG ('day 0') and sister-chromatid exchanges (SCEs) or high frequency cells (HFC) measured 5 days post-exposure. At day 21 post-exposure, SCEs and HFCs in-part persisted and were significantly correlated with persistent N7-HEG adducts. No statistically significant dose effect relationships were observed for induction of micronuclei, nor for chromosome breaks or translocations. In conclusion, this study indicates that following sub-chronic exposure, EO is only weakly mutagenic in adult rats. Using the data of this study to predict cancer risk in man resulting from low level EO exposures in conjunction with other published data, i.e., those on (a) genotoxic effects of EO in humans and rats, (b) DNA binding of other carcinogens, (c) natural background DNA binding and (d) genotoxic potency of low energy transfer (LET) radiation, it is not expected that long term occupational exposure to airborne concentrations of EO at or below 1 ppm EO produces an unacceptable increased risk in man. PMID- 10686306 TI - DNA repair: models for damage and mismatch recognition. AB - Maintaining the integrity of the genome is critical for the survival of any organism. To achieve this, many families of enzymatic repair systems which recognize and repair DNA damage have evolved. Perhaps most intriguing about the workings of these repair systems is the actual damage recognition process. What are the chemical characteristics which are common to sites of nucleic acid damage that DNA repair proteins may exploit in targeting sites? Importantly, thermodynamic and kinetic principles, as much as structural factors, make damage sites distinct from the native DNA bases, and indeed, in many cases, these are the features which are believed to be exploited by repair enzymes. Current proposals for damage recognition may not fulfill all of the demands required of enzymatic repair systems given the sheer size of many genomes, and the efficiency with which the genome is screened for damage. Here we discuss current models for how DNA damage recognition may occur and the chemical characteristics, shared by damaged DNA sites, of which repair proteins may take advantage. These include recognition based upon the thermodynamic and kinetic instabilities associated with aberrant sites. Additionally, we describe how small changes in base pair structure can alter also the unique electronic properties of the DNA base pair pi stack. Further, we describe photophysical, electrochemical, and biochemical experiments in which mismatches and other local perturbations in structure are detected using DNA-mediated charge transport. Finally, we speculate as to how this DNA electron transfer chemistry might be exploited by repair enzymes in order to scan the genome for sites of damage. PMID- 10686307 TI - Population risk and physiological rate parameters for colon cancer. The union of an explicit model for carcinogenesis with the public health records of the United States. AB - The relationship between the molecular mechanisms of mutagenesis and the actual processes by which most people get cancer is still poorly understood. One missing link is a physiologically based but quantitative model uniting the processes of mutation, cell growth and turnover. Any useful model must also account for human heterogeneity for inherited traits and environmental experiences. Such a coherent algebraic model for the age-specific incidence of cancer has been developing over the past 50 years. This development has been spurred primarily by the efforts of Nordling [N.O. Nordling, A new theory on the cancer-inducing mechanism, Br. J. Cancer 7 (1953) 68-72], Armitage and Doll [P. Armitage, R. Doll, The age distribution of cancer and a multi-stage theory of carcinogenesis, Br. J. Cancer 8 (1) (1954) 1-12; P. Armitage, R. Doll, A two-stage theory of carcinogenesis in relation to the age distribution of human cancer, Br. J. Cancer 9 (2) (1957) 161 169], and Moolgavkar and Knudson [S.H. Moolgavkar, A.G. Knudson Jr., Mutation and cancer: a model for human carcinogenesis. JNCI 66 (6) (1981) 1037-1052], whose work defined two rate-limiting stages identified with initiation and promotion stages in experimental carcinogenesis. Unfinished in these efforts was an accounting of population heterogeneity and a complete description of growth and genetic change during the growth of adenomas. In an attempt to complete a unified model, we present herein the first means to explicitly compute the essential parameters of the two-stage initiation-promotion model using colon cancer as an example. With public records from the 1930s to the present day, we first calculate the fraction at primary risk for each birth year cohort and note historical changes. We then calculate the product of rates for n initiation mutations, the product of rates for m promotion-mutations and the average growth rate of the intermediate adenomatous colonies from which colon carcinomas arise. We find that the population fraction at primary risk for colon cancer risk was historically invariant at about 42% for the birth year cohorts from 1860 through 1930. This was true for each of the four cohorts we examined (European- and African-Americans of each gender). Additionally, the data indicate an historical increase in the initiation-mutation rates for the male cohorts and the promotion mutation rates for the female cohorts. Interestingly, the calculated rates for initiation-mutations are in accord with mutation rates derived from observations of mutations in peripheral blood cells drawn from persons of different ages. Adenoma growth rates differed significantly between genders but were essentially historically invariant. In its present form, the model has also allowed us to calculate the rate of loss of heterozygosity (LOH) or loss of genomic imprinting (LOI) in adenomas to result in the high LOH/LOI fractions in tumors. But it has not allowed us to specify the number of events m required during promotion. PMID- 10686308 TI - Somatic mutations and aging: a re-evaluation. AB - Aging has been explained in terms of an accumulation of mutations in the genome of somatic cells, leading to tissue atrophy and neoplasms, as well as increased loss of function. Recent advances in transgenic mouse modeling and genomics technology have created, for the first time, the opportunity to begin testing this theory. In this paper the existing evidence for a possible role of somatic mutation accumulation in aging will be re-evaluated on the basis of the evolutionary logic of aging and recent insights in genome structure and function. New strategies for investigating the relationship between genome instability, mutation accumulation and aging will be discussed. PMID- 10686309 TI - Nephrotoxicity of calcineurin inhibitors: new therapeutic approaches. PMID- 10686310 TI - Mycophenolate mofetil monotherapy: an example of a safe nephrotoxicity/atherogenicity-free immunosuppressive maintenance regimen in a selected group of kidney-transplanted patients. PMID- 10686311 TI - Nephrotoxicity-free, mycophenolate mofetil-based induction/maintenance immunosuppression in elderly recipients of renal allografts from elderly cadaveric donors. PMID- 10686312 TI - Protocol core biopsy (CADI): a surrogate marker for chronic rejection. Mycophenolate Mofetil ICM 1866 and IICR 023 Study Groups. PMID- 10686313 TI - Is tolerance a clinical reality? PMID- 10686314 TI - Steroid withdrawal in renal transplant recipients. PMID- 10686315 TI - The influence of environmental contaminants on lysosomal activity in the digestive cells of mussels (Mytilus galloprovincialis) from the Venice Lagoon. AB - Lysosomes are subcellular organelles bounded by a semipermeable lipoprotein membrane that contain a battery of hydrolytic enzymes that are collectively capable of degrading all classes of indogenous and exogenous macromolecules. Lysosomes accumulate a diverse range of chemical contaminants which can lead to membrane damage resulting in leakage of their contents into the cytosol and damage to cells. Total lysosomal activity for two acid hydrolases, N-acetyl-beta D-hexosaminidase and beta-glucuronidase, with different substrate specificities was determined histochemically in digestive gland sections of mussels, Mytilus galloprovincialis from a series of sites in the Venice Lagoon and the Adriatic Sea and correlated, using multi-stepwise regression analysis, with tissue contaminant burdens in order to explore causality. The results indicated that whilst activity of N-acetyl-beta-D-hexosaminidase correlated with body burdens of mercury, beta-glucuronidase, by contrast, correlated with DDT, Arochlor 1254 and eight PCB congeners in combination with iron or zinc. PMID- 10686316 TI - Estrogenic response of bisphenol A in rainbow trout (Oncorhynchus mykiss). AB - Bisphenol A (BPA) previously shown to possess xenoestrogenic activities was administered to rainbow trout (Oncorhynchus mykiss) through a continuos flow system. The estrogenic response expressed as the induction of vitellogenin (VTG) synthesis was measured during 12 days of exposure, using a direct sandwich ELISA. Quantification of internal liver and muscle concentrations of non-metabolised BPA was performed by LC-MS at the end of the exposure period. A significant induction of the VTG synthesis was obtained at 500 ug BPA/l exposure, although an increase in the ratio of responding animals was observed already between 40 and 70 ug BPA/l. An increase in VTG levels was observed for the 500 ug BPA/l group over the study period, whereas constant or decreasing levels could be detected in the low exposure groups between days 6 and 12. Average internal liver concentrations of BPA increased from 0.22 to 4.36 ug/g for the 10-500 ug BPA/l groups. However, BPA could not be detected in muscle tissue below an exposure level of 70 ug BPA/l. A dose response relationship was established between the internal liver concentrations of BPA and the corresponding VTG responses, with a P<0.001 and a correlation coefficient of 0.66. PMID- 10686317 TI - Relationship between toxicant transfer kinetic processes and fish oxygen consumption. AB - Three organic compounds of different hydrophobicity, 1,2,4,5-tetrachlorobenzene (TeCB), 3,4,5,6-tetrachloroguaiacol (TeCG) and 4,6-dichlorobenzenediol (DBD), were chosen as the test chemicals to carry out a series of investigations to look at the relationship between toxicant transfer and fish metabolic rate. A significant correlation was found between the toxicant uptake rate constant (k(1)) and fish oxygen consumption, regardless of fish size and species. This correlation was improved when fish toxicant body load was expressed on a percent body lipid basis. Similarly, there also existed a significant relationship between the toxicant depuration rate constant (k(2)) and fish oxygen uptake for a range of chemicals with different octanol/water partition coefficients (K(ow)). PMID- 10686318 TI - A physiological model to predict xenobiotic concentration in fish. AB - A physiological model was developed to estimate fish body toxicant load based on information regarding the chemical exposure regime, fish body weight, lipid content and oxygen uptake. The general model was tested in which an oxygen database (OXYREF) was used to predict fish toxicant body burden. Based on the quantitative analysis, it was shown that the model was reliable and accurate in estimating fish body burden of a number of non-metabolized aquatic toxicants. This modified model possesses some functional reality which enables more realistic predictions, making it useful in the practice of aquatic environmental risk assessment. PMID- 10686319 TI - Down-regulation of fibronectin in rainbow trout gonadal cells exposed to retinoic acid. AB - Exposure of fish to some environmental contaminants results in alterations to the levels of retinoid (Vitamin A) stores, which could result in an increase in cellular concentrations of biologically active metabolites such as retinoic acid (RA). However, a link has not been established between changes in retinoid metabolism and impacts on the health of biota. In vitro studies with mammalian cells have demonstrated a relationship between exposure to RA and expression of the extracellular matrix protein, fibronectin (FN); a protein critical for cell migration, adhesion, and transformation. In this study, in vitro exposures of rainbow trout gonadal cells (RTG-2) to RA reduced levels of FN in culture medium; as measured using SDS-PAGE and immunoblot analysis with antisera prepared against RTG-2 cellular fibronectin. This apparent down-regulation of FN secretion occurred in a dose-dependent manner over a range of RA concentrations (10(-10) 10(-6) M). FN down-regulation was not accompanied by changes in the morphology of RTG-2. Future studies should be directed at determining the relationships between retinoid metabolism and FN expression and the potential effects of contaminant induced changes to vitamin A metabolism on the health of fish. PMID- 10686320 TI - The sensitivity of grass shrimp, Palaemonetes pugio, embryos to organophosphate pesticide induced acetylcholinesterase inhibition. AB - Grass shrimp, Palaemonetes pugio, are common inhabitants of salt marshes along the Atlantic and Gulf coasts of North America. Grass shrimp embryos are brooded externally on the abdomen of adult females for about 2 weeks prior to hatching. In South Carolina, the spring spawning period for grass shrimp coincides with the period of peak pesticide application on crops grown along the South Carolina coast. Thus, grass shrimp of all developmental stages are at risk of exposure to pesticides present in nonpoint source agricultural runoff. Organophosphate (OP) insecticides are commonly applied agricultural chemicals which produce toxicity by inhibiting the nervous system enzyme, acetylcholinesterase (AChE). The purpose of this study was to examine the development of AChE activity in grass shrimp embryos and to assess their sensitivity to OP-induced AChE inhibition. Embryos were exposed for 24 h to either chlorpyrifos or malathion. All exposure concentrations were nominal and ranged from 0 to 2.00 ug l(-1) for chlorpyrifos and from 0 to 120.00 ug l(-1) for malathion. Quantifiable levels of AChE activity first appeared at Stage V of development and increased as embryonic development progressed. AChE inhibition by the OPs was assessed in Stage VI and Stage VII embryos. Both stages of embryos were more sensitive to chlorpyrifos than malathion. The 24-h Effective Concentration (EC(50)) values for chlorpyrifos were 0.49 ug l(-1) (95% C.I.=0.33-0.77 ug l(-1)) and 0.36 ug l(-1) (95% C.I.=0.33-0.38 ug l(-1)) for Stage VI and Stage VII embryos, respectively. In comparison, malathion 24-h EC(50) values were 55.53 ug l(-1) (95% C.I.=22.08-80.73 ug l(-1)) for Stage VI embryos and 29.93 ug l(-1) (95% C.I.=25.22-44.22 ug l(-1)) for Stage VII embryos. For both OPs, there were no significant differences in the EC(50) values calculated for Stage VI and Stage VII embryos; however, AChE inhibition was significantly (P NR1/2D > NR1/2C > or = NR1/2A. Our observations may provide an explanation for the reduced neurotoxicity of high doses of gp120 in cell cultures and may be useful for the pharmacological discrimination of NMDA receptor subtypes. PMID- 10686401 TI - Effects of morphine glucuronides on the function of opioid receptors in human SK N-SH cells. AB - Morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) are active metabolites of morphine. The effects of M3G and M6G on the opioid receptor transduction system has not yet been fully elucidated. Formation of cAMP after treatment with various doses of morphine, M3G, and M6G was studied. M6G and morphine, but not M3G, showed a dose dependent inhibition of cAMP accumulation. Naloxone blocked the inhibitory effect of M6G, M3G, and morphine. Pretreatment with M3G did not change the effects of morphine and M6G. The G-protein inhibitor PTX, prevented morphine, M3G, and M6G effects on cAMP. M3G and M6G vary in their ability to interact with the opioid receptor effector system. Inhibition of cAMP evoked by activation of opioid receptors and inhibitory G-proteins may play a role in the actions of M6G and M3G. PMID- 10686402 TI - Potassium currents in CA1 neurons of rat hippocampus increase shortly after transient cerebral ischemia. AB - Total potassium current in CA1 pyramidal neurons was studied using whole-cell voltage-clamp recording technique in hippocampal slices prepared before and at 6 8 h after transient forebrain ischemia. The total potassium current significantly increased from a control value of 2.17+/-0.17 to 2.96+/-0.31 nA (measured at +70 mV, P<0.05) after ischemia. The slope factor V(c) of activation curve significantly decreased and the half-inactivation voltage V(h) shifted to more depolarized potentials after ischemia. These results indicate that the increase of potassium current might be responsible for the decreased excitability in CA1 neurons after severe ischemia and may be involved in postischemic cell death in hippocampus. PMID- 10686403 TI - Correlation between behavior and extracellular dopamine levels in rat striatum: comparison of microdialysis and fast-scan cyclic voltammetry. AB - Recently, fast-scan cyclic voltammetry (FSCV) has been adapted for real-time measurements of evoked dopamine (DA) release and uptake in freely moving rats. Using the advantages of this experimental design in combination with behavioral measures, we examined the effect of GBR 12909 (20 mg/kg, i.p.), a selective DA uptake inhibitor, on striatal extracellular DA dynamics and compared these data to that obtained by microdialysis. These studies established that both techniques report changes in DA that correlate with the kinetics of GBR 12909-induced behavioral effects. However, the time course of changes in evoked DA levels detected by FSCV was more closely linked with the changes in stereotypy than microdialysis measurements. PMID- 10686404 TI - Expression of insulin-like growth factor-I in rat glioma cells is associated with change in both immunogenicity and apoptosis. AB - Insulin-like growth factor I (IGF-I), has a role in cellular differentiation and is also expressed in neoplastic transformation of glioma cells. We recently demonstrated inhibition in expression of cellular IGF-I after transfection with vectors that incodes a segment of the human IGF-I RNA in antisense orientation. The transfected cells expressed increased levels of both MHC-I and B7 molecules. In this paper we show that IGF-I antisense transfected cells also become apoptotic. Moreover, the phenomenon of programmed cell death is related to the phenomenon that results in increased expression of MHC-I and B7 molecules. Co transfection of rat glioma cells with the vector expressing IGF-I antisense RNA and with vectors encoding the expression of MHC-I and B7 antisense cDNA suppressed the expression of both of these molecules and was associated with a decrease in apoptosis. PMID- 10686405 TI - Amphetamine-induced zif268 mRNA expression in the medial posterior nucleus accumbens in cholecystokinin-A receptor mutant rats. AB - Converging evidence supports a role for cholecystokinin (CCK) in modulating dopamine (DA)-mediated activity in the rat mesolimbic system. In particular, CCK co-localized with mesolimbic DA cells originating in the ventral tegmental area potentiates DA function in the medial posterior nucleus accumbens (mpNA) through CCK-A receptors. Recently, a strain of rats lacking the CCK-A receptor, Otsuka Long Evans Tokushima Fatty (OLETF), has been discovered making it possible to study the mesolimbic DA regulatory role of CCK-A receptors. Previous studies have shown that OLETF rats are less sensitive to amphetamine (AMPH)-induced behavioral effects compared to controls. To determine if this altered sensitivity is associated with decreased AMPH-induced postsynaptic activation in the mpNA in OLETF rats, we performed the following experiment. OLETF (CCK-A mutants) and Long Evans Tokushima Otsuka (LETO) rats (controls) were given subcutaneous injections of either saline or AMPH (5.0 mg/kg). One hour after injection all animals were sacrificed and activation of the mpNA was assessed using in situ hybridization with antisense probes for zif268 mRNA. AMPH treatment produced a significant up regulation of zif268 mRNA expression in both OLETF and LETO rats (P/=0.25), and all but one child started language with a preference for content words as opposed to function words. PMID- 10686432 TI - Pituitary prolactin-secreting tumor formation: recent developments. AB - Prolactinoma is the most common type of primary pituitary tumors. It occurs more frequently in women than in men. Dopaminergic agonists are effective in the shrinkage of prolactin-secreting pituitary tumor and are preferred in some patients. However, pituitary radiotherapy may enable the long-term removal of prolactin-secreting tumor cells. Recent evidence suggests that prolactinoma is a heterogeneous disorder with complicated and multifactorial etiology and pathogenesis. Apparently, a thorough understanding of prolactinoma tumorigenesis would be important. To facilitate investigations on tumorigenesis of prolactinoma, animal models for prolactinomas have been developed. These models have expedited our progress in the recent years. Many researchers consider the F(344) rat to be the most sensitive strain of rats to estrogen (E(2))-induced prolactinoma formation. Nonetheless, E(2) treatment for 60 days also induces the formation of pituitary prolactin-secreting adenoma in male Sprague-Dawley (SD) rats. Evidently, the SD rat is also a good animal for prolactinoma investigations. Following E(2) implantation, prolactinomas developed in the eutopic adenohypophysis in situ and/or ectopic pituitary grafted under the renal capsule in SD rats. These observations favor the hypothesis that prolactinoma growth is the result of pathological changes in the adenohypophysis and/or hypothalamus. In the latter case, abnormal release of hypothalamic dopamine, GABA, or brain-gut peptides (such as cholecystokinin, vasoactive intestinal polypeptide, galanin, angiotensin, opioid peptide, gastrin, gastrin-releasing peptide, pancreatic polypeptide, and adrenocorticotropic hormone) results in some of the pathological changes that may lead to hyperprolactinemia and/or prolactinoma development. Dysregulation of prolactin synthesis and secretion may be the result of prolactin gene modulation. In E(2)-induced rat prolactinomas, prolactin mRNA contents and the expression of some proto-oncogenes, e.g. c-myc and c-ras, TGFalpha and TGFbeta1 mRNA were significantly changed. The above findings are consistent with results in human prolactinoma development. In addition, in rats abnormal expression of the prolactin gene was correlated with hypomethylated status of CpG sites in exons 1, 2 and 4 of the prolactin gene, as well as the increase in hypersensitive sites to DNase 1 in the encoding region of the prolactin gene. In E(2)-treated rats, a point mutation with a base substitution from cytidine (C) to adenine (A) was found at the -36-bp site of the proximal promoter of the prolactin gene in eutopic pituitary prolactinomas, but no change was observed in the same sequence of the prolactin gene in ectopic prolactinoma. The association of a base substitution with the hyperexpression of the prolactin gene in eutopic prolactinomas suggests that different mechanisms may mediate the formation of eutopic and ectopic prolactin-secreting tumors. Melatonin decreases the expression of the prolactin gene in vitro suggesting that this pineal hormone may be a potential anticarcinogen in vivo. It has also been shown that MT(2) (Mel(1b)) melatonin receptors are expressed in anterior pituitary cells. The use of melatonin as a preventive or therapeutic drug for prolactinomas should be further investigated. In summary, improved knowledge on tumorigenesis of prolactinomas, especially in the rat model, was noted. These E(2)-induced rat prolactinoma models would facilitate future investigations, and expected results shall be fruitful and exciting for the development of future drug designs for the prevention and/or treatment of prolactin-secreting pituitary tumors. PMID- 10686433 TI - The effect of protein kinase C activation on G(z)-mediated regulation of type 2 and 6 adenylyl cyclases. AB - Three serine-to-alanine mutants of the alpha subunit of the heterotrimeric G protein G(z) (alpha(z)) were examined for their signaling properties in the presence of phorbol ester treatment. All three alpha(z) mutants resembled wild type alpha(z) in their abilities to inhibit alpha(s)-stimulated type 6 adenylyl cyclase (AC6) and phorbol ester treatment reduced their magnitudes of inhibition. Depending on the permissive condition, the betagamma-mediated stimulation of type 2 adenylyl cyclase (AC2) was differentially regulated by alpha(z) and the three mutants. Mutation of Ser(27) but not Ser(16) of alpha(z) affected the efficient release of betagamma subunits upon receptor activation and abolished the stimulation of phosphorylated but not alpha(s)-stimulated AC2. PMID- 10686434 TI - Changes in leucine uptake in the retina of the hamster after traumatic detachment. AB - Protein metabolism was investigated in detached hamster retinas. By sucking off 0.2 ml of aqueous humor from the anterior chamber through limbic insertion of a 27-gauge needle, a tractional force pulled off the neural retina from the retinal pigment epithelium and created a simple detachment without retinal breaks in the right eyes of the hamsters. The left eyes were left untouched as normal controls and sham controls were induced by simple limbic insertion without suction. The animals were sacrificed at selected intervals of 1, 3, 6, 9, 16, 24, 32 days after the operation. Subsequently, scintillation counting and autoradiography were employed to study retinal protein metabolism using leucine uptake as an index. After tritiated leucine uptake, scintillation counting of radioactive substance indicated that detached retinas had taken in less tritiated leucine than normal controls from day 1 to 6 after the operation, but this change had normalized by day 9. For autoradiography, the change in leucine uptake rate was shown to be different in different layers. All the retinal cells seemed to show a decreased leucine uptake with the exception of the outer nuclear layer, in which leucine appeared to be significantly upregulated. This paper illustrates the patterns of protein metabolism and their change after traumatic detachment as well as their possible recovery. PMID- 10686435 TI - Projection linkage from spinal neurons to both lateral cervical nucleus and solitary tract nucleus in the cat. AB - Intracellular recordings from the lumbosacral dorsal horn were made to identify the axonal projection and the afferent innervation of the lateral cervical nucleus (LCN) and solitary tract nucleus (STN) on the spinal neurons of chloralose-anesthetized cats. A total of 49 neurons from laminae III-V in the spinal dorsal horn responded to stimulation of both the LCN and STN. Of these, 28 and 21 neurons responded antidromically and orthodromically to stimulation of the LCN and STN, respectively. Seven of the 28 antidromically activated neurons were followed by one or more responses synaptically driven from the LCN and/or STN. The diameter of these ascending or descending fibers was in the range of A delta fibers. The results indicate that (1) some spinal neurons, namely spinocervical tract-spinosolitary tract (SCT-SST) neurons, issue branched axons of A delta fibers and dually project to both LCN and STN; (2) some SCT-SST neurons receive innervation from both the LCN and STN; (3) some spinal neurons and interneurons are dually innervated by descending fibers originating from both the LCN and STN, and (4) the convergence and integration between somatic and visceral sensory inputs might occur in the SCT-SST neurons. PMID- 10686436 TI - Purification of rat C6 glioma plasma membranes by affinity partitioning. AB - We have studied the feasibility of purifying rat C6 glioma plasma membranes by a phase partitioning approach. The purification procedure involves cell homogenization and fractionation with an aqueous two-phase polymer system followed by selective affinity purification of plasma membranes by a wheat germ agglutinin-coupled polymer system. We demonstrate that the two-phase affinity partitioning technique is a simple and efficient method of isolating cell plasma membranes with high purity and yield. Furthermore, the isolated plasma membranes retain their functional integrity, as shown by the high-affinity insulin-like growth factor-I (IGF-I) binding capacity of IGF-I receptors. PMID- 10686437 TI - The role of dialyzer biocompatibility in acute renal failure. PMID- 10686438 TI - Impaired biological activity of erythropoietin by cyanate carbamylation. AB - AIMS: During advanced renal failure, particularly in patients with end-stage renal disease (ESRD), proteins are carbamylated as a result of a reaction with cyanate. Some or all of the cyanate is derived from urea. If the carbamylation of proteins adversely alters their biologic activities, then urea must be viewed as an uremic toxin, rather than a surrogate. Therefore, we studied the effect of cyanate carbamylation on the erythropoietic activity of erythropoietin (EPO) in a rodent model. METHODS: EPO was carbamylated by incubation with cyanate at 37 degrees C. The extent of carbamylation was monitored using trinitrobenzenesulfonic acid. In Sprague-Dawley rats the erythrocyte count, hemoglobin concentration, and hematocrit were measured after the twice-weekly subcutaneous injection of either EPO or carbamylated EPO for 3 weeks. Two additional control groups received physiologic saline or 0.2 ml of 1 M cyanate. RESULTS: The level of carbamylated EPO was increased as the time of exposure to cyanate increased from 1 to 6 h, and as the cyanate concentration increased from 8 to 2,000 mM. EPO injections caused significantly large increases in all erythropoietic measures. Physiologic saline or 1 M cyanate-injected controls and the carbamylated EPO-injected animals demonstrated no change from baseline in erythropoietic parameters. CONCLUSION: These results support that EPO exposed to high levels of cyanate in vitro demonstrates diminished biologic activity in healthy Sprague-Dawley rats. This effect may be manifested by the carbamylation of EPO by the cyanate. Should this occur in ESRD patients, it may contribute to the suboptimal erythropoietic response to EPO therapy associated with high urea levels, especially related to inadequate dialysis. Targeting dialysis doses specifically to urea concentrations may be more important than previously considered. PMID- 10686439 TI - Analysis of urea nitrogen and creatinine kinetics in hemodialysis: comparison of a variable-volume two-compartment model with a regional blood flow model and investigation of an appropriate solute kinetics model for clinical application. AB - To investigate an appropriate solute kinetics model for clinical application, we analyzed urea nitrogen (UN) and creatinine (Cr) kinetics by a variable-volume two compartmental model (2CM) and a regional blood flow model (RBF) in 44 hemodialysis patients with varying proportions of first compartmental volume and regional volume (p(1)). Solute kinetics could not be solved in some of the patients with higher p(1) values, and there were more solution failures by the RBF than by the 2CM. The solute generation rate (g) and solute distribution volume in the dry state (V(D)) increased with increases in p(1) in both models, but there were some differences between the two models. When g was normalized by V(D), it became relatively constant, irrespective of the p(1) value or model used (0.133 +/- 0.029 mg/min/l by the 2CM and 0.132 +/- 0.029 mg/min/l by the RBF for UN; 0.0200 +/- 0.0049 mg/min/l by the 2CM and 0.0198 +/- 0.0048 mg/min/l by the RBF for Cr). The intercompartmental mass transfer coefficient (K(c); liters/min) calculated by the 2CM decreased as p(1) increased (K(c) = -1.77.p(1) + 1.16, p < 0.0001, R = 0.999 for UN; K(c) = -0.847.p(1) + 0.556, p < 0.0001, R = 1.000 for Cr). The systemic blood flow (Q(sys); liters/min) calculated by the RBF also decreased as p(1) increased (Q(sys) = -11.1.p(1) + 6.21, p < 0.0005, R = 1.000 for UN; Q(sys) = -5.22.p(1) + 2.90, p < 0.001, R = 0.999 for Cr). Since the RBF more frequently failed to solve the solute kinetics and since there was a difference in its Q(sys) values for UN and Cr, the 2CM was considered to be a superior model. When p(1) was extremely low, the 2CM could be transformed into a modified variable-volume one-compartment model (1CM) which presented a similar g/V(D) (0.133 +/- 0.029 for UN; 0.0200 +/- 0.0048 for Cr). This modified 1CM was considered to satisfy appropriate conditions for clinical application, since it is simpler than the 2CM and provides useful information on the dialysis dose. PMID- 10686440 TI - Plasma C-reactive protein in hemodialysis patients: a cross-sectional, longitudinal clinical survey. AB - In hemodialysis patients, C-reactive protein (CRP), an acute-phase reactant, is a sensitive and independent marker of malnutrition, anemia, and amyloidosis. The aim of the present studies was to evaluate CRP and interleukin 6 levels in plasma samples from long-term hemodialysis patients on different extracorporeal modalities associated with or without backfiltration. Two hundred and forty-seven patients were recruited in eight hospital-based centers. All patients had been on their dialytic modality for at least 6 months. At enrollment, 46 hemodialysis patients out of 247 (18.6%) had clinical evidence of pathologies known to be associated with high CRP values. The 201 remaining patients were defined as clinically stable and were on conventional hemodialysis (34%), hemodiafiltration with infusion volumes <10 liters/session (10%), hemodiafiltration with infusion volumes <20 liters/session (32%), and double-chamber hemodiafiltration with infusion volumes <10 liters/session (22%). Analysis of CRP values in the clinically stable patients showed that an unexpectedly high proportion (47%) of the patients had CRP values higher than 5 mg/l (taken as the upper limit in normal human subjects). The values of CRP and interleukin 6 were significantly higher in hemodiafiltration with infusion volumes <10 liters/session than in hemodiafiltration with infusion volumes >20 liters/session, in hemodialysis and in double-chamber hemodiafiltration. The same pattern occurred after 6 months of follow-up in 171 out of 201 clinically stable patients. Hemodialytic conditions that expose to the risk of backfiltration such as low exchange volume hemodiafiltration may induce a chronic inflammatory state as reflected by increased plasma values of both CRP and interleukin 6, thus suggesting the need for hemodialytic strategies that reduce (hemodialysis with low-permeability membranes or hemodiafiltration with infusion volumes >20 liters) or eliminate (double-chamber hemodiafiltration) backfiltration of bacteria-derived contaminants. PMID- 10686441 TI - Evaluation of intradialytic solute and fluid kinetics. Setting Up a predictive mathematical model. AB - A mathematical model of solute kinetics for the improvement of hemodialysis treatment is presented. It includes a two-compartment description of the main solutes and a three-compartment model of body fluids (plasma, interstitial and intracellular). The main model parameters can be individually assigned a priori, on the basis of body weight and plasma concentration values measured before beginning the session. Model predictions are compared with clinical data obtained in vivo during 11 different hemodialysis sessions performed on 6 patients with a profiled sodium concentration in the dialysate and a profiled ultrafiltration rate. In all cases, the agreement between the time pattern of model solute concentrations in plasma and the in vivo data proves fairly good as to urea, sodium, chloride, potassium and bicarbonate kinetics. Only in two sessions was blood volume directly measured in the patient, and in both cases the agreement with model predictions was good. In conclusion, the model allows a priori computation of the amount of sodium removed during hemodialysis, and makes it possible to predict the plasma volume changes and plasma osmolarity changes induced by a given sodium concentration profile in the dialysate and by a given ultrafiltration profile. Hence, it can be used to improve clinical tolerance to the dialysis session taking the characteristics of individual patients into account, in order to minimize intradialytic hypotension. PMID- 10686442 TI - Effect of plasma exchange on serum tissue inhibitor of metalloproteinase 1 and cytokine concentrations in patients with fulminant hepatitis. AB - AIMS: The present study assessed whether the serum concentrations of tissue inhibitor of metalloproteinase 1 (TIMP-1) and cytokines are altered in patients with fulminant hepatitis and whether plasma exchange affects these concentrations. METHODS: Fifteen patients with fulminant hepatitis, 14 patients with severe acute hepatitis, and 20 healthy controls were included in this study. The serum levels of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), interleukin 6 (IL-6), transforming growth factor beta (TGF-beta), and TIMP-1 were determined in all patients upon hospital admission and before and after a single course of plasma exchange in the patients with fulminant hepatitis. RESULTS: Ten out of the 15 patients with fulminant hepatitis and all patients with severe acute hepatitis survived. Serum TNF-alpha, IL-6, TGF-beta, and TIMP-1 levels in patients with fulminant hepatitis were significantly higher than the levels in patients with severe acute hepatitis (p < 0.01). IL-1beta was not detectable in either group. Plasma exchange reduced the increased serum concentrations of TNF-alpha, IL-6, TGF-beta, and TIMP-1 in patients with fulminant hepatitis (p < 0.01). CONCLUSIONS: These data suggest that increased serum levels of TIMP-1 and cytokines may reflect severe hepatic inflammation and that plasma exchange is an effective therapy to reduce these levels. PMID- 10686443 TI - Clinical pharmacokinetics and effects of an oral sustained-release preparation of disopyramide prescribed for patients undergoing maintenance hemodialysis. AB - AIMS: We evaluated the clinical pharmacokinetics of a sustained-release preparation of disopyramide phosphate (DSR) and its effects on supraventricular arrhythmias in hemodialysis patients. METHODS: Eight hemodialysis patients with either paroxysmal supraventricular tachycardia (PSVT) or PSVT plus paroxysmal atrial fibrillation (Paf) were given 150 mg of DSR 2 h before each hemodialysis. The frequency of PSVT, the duration of Paf before and 2 weeks after starting DSR and the blood concentration of the drug were evaluated. RESULTS: There was no significant difference between serum levels of DSR before and after hemodialysis. The frequency of PSVT and the duration of Paf were significantly reduced by the therapy. Side effects and electrocardiographic abnormalities did not appear during the period. CONCLUSION: We conclude that hemodialysis does not remove DSR, and that a single dose of 150 mg of DSR given 2 h before hemodialysis is safe and sufficient to reduce the incidence of supraventricular arrhythmias. PMID- 10686444 TI - Salt and hypertension in end-stage renal disease. PMID- 10686445 TI - Cholesterol reduction and stroke occurrence: an overview of randomized clinical trials. AB - We performed a meta-analysis of randomized clinical trials of more than 6 months duration to describe how fatal and nonfatal strokes are related to cholesterol lowering and to the type of intervention. A total of 41 individual trials including approximately 80,000 subjects and followed for an average of about 4 years were included in the overview. There was a 16% (95% CI, 7-25%) reduction in risk of stroke among treated patients compared to control patients (test for heterogeneity, p = 0.76). When trials that used different interventions were separately examined, a significant reduction in stroke occurrence was observed only for those using statins as active treatment (risk reduction 23%; 95% CI 13 33%). A variance-weighted regression analysis of the logarithmic odds ratios for stroke incidence against the percentage of cholesterol reduction indicated that a reduction of fatal and nonfatal stroke can be obtained for a cholesterol reduction of 9% (95% CI 6.8-13.6%). The combined data of primary and secondary prevention trials indicate that a large reduction of blood cholesterol, achievable with statin drugs, can reduce the incidence of stroke. PMID- 10686446 TI - Clinician's biases toward surgery in cerebellar hematomas: an analysis of decision-making in 94 patients. AB - BACKGROUND AND PURPOSE: No studies have examined clinical decision-making in cerebellar hemorrhages. Clinical and CT features may influence surgery in patients with a spontaneous cerebellar hematoma. One commonly accepted adage is to remove a clot when 3 cm or larger in axial diameter on presentation CT scan. It is possible that certain preferences impact on outcome. METHODS: We analyzed 94 patients with spontaneous cerebellar hematomas between the years of 1973-1993. Thirty-one patients underwent suboccipital craniectomy and clot removal with or without ventriculostomy. Deterioration denoted worsening of consciousness, new brainstem signs, or presentation in coma. Statistical analysis was performed utilizing a tree-based model fitted by binary recursive partitioning. Ninety-five percent confidence intervals (CI) were calculated. RESULTS: The overall probability of surgical intervention was 33% (CI 23-43%). The chance of surgery in stable patients was 7% (CI 2-20%). Neurologic deterioration was seen in 54 patients (57%) and increased the prospects of a surgical procedure (52%, CI 38 66%). Surgery was performed in all deteriorating patients with small hematomas (size <3 cm), but large clots (size >3 cm) did not substantially influence surgical decision-making (45%, CI 30-60%) except in patients younger than 70 years old (57%, CI 41-82%). CONCLUSIONS: Clinicians at our institution usually wait for clinical deterioration to unfold prior to operating on patients with cerebellar hematomas. After deterioration occurs, they prefer small hematomas but will operate on large hematomas when patients are younger than 70, generally withholding surgery from older patients. These attitudes may impact on outcome and should be considered in future treatment trials. PMID- 10686448 TI - Distribution, severity and risk factors for aortic atherosclerosis in cerebral ischemia. AB - Significant thoracic aortic plaques (>4 mm) are an independent risk factor for ischemic stroke. Within 1 week of stroke/transient ischemic attack (TIA) onset, 105 consecutive patients underwent transesophageal echocardiography assessment of aortic plaque thickness using the criteria of Amarenco et al. (N Engl J Med 1994;331:1474-1479). A proximo-distal gradient was found in the distribution of aortic atheroma >4 mm (p = 0.04). Symptomatic coronary artery disease was associated with plaque in the proximal aorta (p = 0.03); extracranial carotid stenosis >70% was associated with plaque in the arch and descending aorta (p < 0.01). The severity of aortic plaque was associated with age on multivariable analysis (p = 0.0003 to p < 0.01). Only smoking showed predictive regional specificity (p = 0.03);no other risk factors were associated with aortic atheroma in any segment. In stroke/TIA patients, carotid stenosis >70% predicts aortic arch atheroma plaques >4 mm which may predispose to reinfarction after endarterectomy. Atheroma of the ascending aorta is associated with ischemic heart disease, and cardiac screening should be considered in asymptomatic patients. PMID- 10686447 TI - Leukocyte activation: relation to cardiovascular mortality after cerebrovascular ischemia. AB - Activated leukocytes are believed to be involved in the pathogenesis and progression of atherosclerotic vascular disease and its consequences. In a 4-year observational follow-up study, we investigated whether markers for systemic leukocyte activation (leukocyte-derived inflammatory mediators) were related to cardiovascular mortality after cerebrovascular ischemia. Using enzyme-linked immunosorbent assays, we measured the plasma levels of soluble tumor necrosis factor receptor protein-1 (sTNFR-1), neutrophil gelatinase-associated lipocalin (NGAL) and neutrophil protease-4 (NP4) in 144 patients (90 stroke, 54 transient ischemic attack) 1-3 days after cerebral ischemia. During the 4 years of follow up, 42 (29%) of the 144 patients died; 38 of cardiovascular causes and 4 of other causes. Patients with evidence of higher leukocyte activation (n = 47) had a higher 4-year cardiovascular mortality rate than those without evidence of leukocyte activation (n = 97; p < 0.005). Logistic regression analysis with age, sex and other significant predictors as covariates showed higher plasma levels of sTNFR1 and NGAL both to be significant independent predictors of cardiovascular mortality, the respective odds ratio, 95% confidence intervals, and p values being 2.0, 1.2-3.4, p < 0.01, and 3.6, 1.2-10.5, p = 0.02, respectively. We concluded that in patients with acute cerebral ischemia, plasma markers of leukocyte activation were significant predictors of long-term cardiovascular mortality. This may indicate an important role of activated leukocytes in the progression of these diseases. PMID- 10686449 TI - Lipoprotein(a), other lipoproteins and hemostatic profiles in patients with ischemic stroke: the relation to cardiogenic embolism. AB - Lipoprotein and hemostatic profiles including coagulation inhibitors were determined in 136 patients with acute ischemic stroke. Based on clinical examination, cerebral computed tomography, Doppler ultrasonography of precerebral arteries and transthoracic echocardiography, the strokes were classified as cardioembolic (n = 38), non-cardioembolic (n = 92), and mixed cardioembolic/hypertensive (n = 6). Patients with cardioembolic stroke were older than patients with non-cardioembolic stroke. Lipoprotein(a) was higher in the cardioembolic than in the non-cardioembolic group. Lipoprotein(a) was not significantly correlated to the other lipid levels and may represent an independent lipid risk factor. The non-cardioembolic group had higher levels of total cholesterol, triglycerides, total cholesterol/high-density lipoprotein cholesterol ratio, low-density lipoprotein cholesterol, apolipoprotein A1, and apolipoprotein B. The cardioembolic group had higher concentrations of fibrinogen and D-dimer, and lower levels of antithrombin, protein C, protein S and heparin cofactor 2 than the non-cardioembolic group. The differences in the hemostatic profile are consistent with thrombosis due to activated coagulation being more involved in the pathogenesis of cardioembolic than of non-cardioembolic stroke. Lipoprotein(a) seems to be more associated with coagulation markers of thrombosis than with atherosclerosis, whereas the other lipids mainly seem to be risk factors for atherosclerosis. PMID- 10686450 TI - Cross-validation of a model predicting discharge home after stroke rehabilitation. Validating stroke discharge predictors. AB - A new sample of 116 stroke patients was collected in order to validate a logistic regression model, predicting the chances of severely affected stroke patients being discharged home to independent living. The model was found to be accurate in the new sample, especially for those patients who had a high estimated probability of being discharged home. When the dividing line for the predicted probability for discharge home was set at a value of >/=0.5, the positive and negative predictive values were 74 and 73%, respectively. Further modelling resulted in a new extended model including the variables postural stability on admission, cohabiting, age and perceptual impairment on admission that formed the basis for an index predicting discharge home. This index was then validated in the sample of 93 patients that the first developed model was derived from and showed positive and negative predictive values of 85 and 77%, respectively. PMID- 10686451 TI - Dynamic but not static cerebral autoregulation is impaired in acute ischaemic stroke. AB - It remains unclear as to whether dynamic and static cerebral autoregulation (CA) are impaired in acute ischaemic stroke, and whether these changes are related to stroke subtype. This could have important implications with regard to post-stroke prognosis and the management of blood pressure (BP) in the acute post-ictal period. Using transcranial Doppler ultrasonography and non-invasive manipulation of BP, we compared both mechanisms in 61 patients with ischaemic stroke within 96 h of ictus, and 54 age- and sex-matched controls. There was no difference in static and dynamic CA indices between the various stroke subtypes. Combining all stroke subtypes dynamic autoregulation, as measured using thigh cuff release, was significantly impaired in both the affected and non-affected stroke hemispheres compared to controls (mean autoregulation index 4.1 +/- 3.3, 4.8 +/- 3.1 and 6.2 +/- 2.3, respectively, p < 0.05). By comparison static autoregulation, assessed using isometric hand grip and thigh cuff inflation, was not significantly different. In conclusion, dynamic but not static CA appears to be globally impaired in acute ischaemic stroke. This deserves further study and may identify possibilities for therapeutic intervention. PMID- 10686452 TI - The Athens stroke registry: results of a five-year hospital-based study. AB - The advent and wide application of new technology, especially noninvasive techniques, has enabled physicians to more completely investigate and clarify the etiopathogenic mechanisms of stroke. Such data have not been available until recently for Southeastern Europe. In addition, during the last decades, strategies for the modification of risk factors and primary prevention may have changed the prevalence of each subgroup of stroke as well. We investigated 1, 042 consecutive patients who had first strokes, during a period of 5 years (from June 1992 to May 1997) and classified them prospectively based on etiopathogenic mechanisms. Patients with transient ischemic attacks and subarachnoid hemorrhage were excluded. There were 613 male and 429 female patients, with a mean age of 70.2 +/- 11.9 years. Forty-six percent of the patients arrived within 3 h from stroke onset. The probable mechanisms were: large-artery atherosclerosis, 156 (15%); lacunes, 177 (17%); cardioembolic, 335 (32.1%); infarct of unknown cause, 182 (17.5%); miscellaneous causes, 35 (3.3%), and intracerebral hemorrhage (ICH), 157 (15.1%). In the cardioembolic group, nonvalvular atrial fibrillation (NVAF) was the probable cause in 225 patients, especially in patients older than 75 years (65%). The overall hospital mortality was 15.2% (from 0.6% for lacunar stroke to 34% for ICH). In our population, cardioembolism is the most frequent subtype of stroke. NVAF is the most likely source, especially in older patients. PMID- 10686453 TI - Measurement of cerebral circulation time by contrast-enhanced Doppler sonography. AB - We present a new, non-invasive ultrasound method for the measurement of cerebral circulation time. After injection of a galactose-based echo-contrast agent (Levovist) into an antecubital vein, cerebral circulation time is measured as the interval between the beginning of a stable signal enhancement of at least 5 dB in the internal carotid artery and in the ipsilateral internal jugular vein. Both vessels are insonated simultaneously at the mandibular angle using a single 2-MHz range-gated transducer. For this study, 25 healthy volunteers ranging in age from 23 to 55 years (30.1 +/- 7.5 years; mean +/- SD) were examined. Cerebral circulation time was 5.6 +/- 1.7 s without significant side-to-side or sex related differences. Measurement of circulation times offers a new possibility for the employment of echo-contrast agents in functional ultrasound. PMID- 10686454 TI - Design of ESPRIT: an international randomized trial for secondary prevention after non-disabling cerebral ischaemia of arterial origin. European/Australian Stroke Prevention in Reversible Ischaemia Trial (ESPRIT) group. AB - The ESPRIT trial addresses the problem that aspirin, the standard therapy for secondary prevention of vascular complications after a transient ischaemic attack (TIA) or ischaemic stroke of arterial origin, reduces the risk of serious vascular events by only about 13%. Anticoagulants may be an alternative, as these have proved highly efficacious in trials after myocardial infarction and after cerebral ischaemia with atrial fibrillation. After cerebral ischaemia of presumed arterial origin, high-intensity anticoagulation (INR 3.0-4.5) is not safe, but the value of anticoagulation with an INR between 2.0 and 3.0 is still unknown. Secondly, a recent, large trial showed that the combination of aspirin and dipyridamole prevents more major vascular events than aspirin alone, but several earlier trials did not find such an advantage. In ESPRIT, patients with a TIA or minor ischaemic stroke (Rankin grade G transition 96 bp upstream of exon 7 of the telSMN gene, a potential disease-causing mutation in these patients. PMID- 10686496 TI - Allelic frequencies of FBN1 gene polymorphisms and genetic analysis of italian families with Marfan syndrome. AB - The fibrillin gene (FBN1) is the disease locus for Marfan syndrome. This disorder shows a high degree of clinical and allelic heterogeneity. Direct mutation screening has proven difficult and inefficient and at present cannot be utilized for routine analysis. In familial cases linkage analysis represents a useful tool for molecular diagnosis. We have determined the allelic frequencies of 5 polymorphic markers within the FBN1 locus in the Italian population and have successfully employed them for prenatal diagnosis and resolution of clinically equivocal cases. PMID- 10686497 TI - Search for coeliac disease susceptibility loci on 7q11.23 candidate region: absence of association with the ELN17 microsatellite marker. AB - The involvement of HLA genes in the susceptibility to coeliac disease (CD) has been well documented and represents the only consistently observed genetic feature of this multifactorial disease. In the present study, the search for new susceptibility genes has been devoted to a candidate region suggested by the association of CD with Williams syndrome (WS). This genetic disorder is due to a deletion in the 7q11.23 region that includes the elastin (ELN) gene. An increased prevalence of CD in WS patients has been previously reported and a case of CD-WS is also described in the present study. We used the ELN17 microsatellite marker mapped within the ELN gene to look for a possible contribution of this region to the susceptibility to CD. The analysis of 74 Italian CD families provided no evidence of association with the ELN17 marker. PMID- 10686498 TI - Testing mode of inheritance of a candidate mutation at a quantitative trait locus. AB - Here is presented an approach to testing whether the effect of a candidate gene on a quantitative trait is dominant and for testing whether the effect is recessive. The approach uses parental genotype information in nuclear families to adjust for bias due to population admixture. The approach is applicable regardless of the nature of the sampling. The results of an application of the methods to a candidate mutation for diabetic nephropathy are used for illustration. PMID- 10686499 TI - Allelic affinities between the F13A common gene products inferred by the analysis of an (AAAG)n STR polymorphism within the 5' untranslated region. AB - Factor XIII a subunit (F13A) is the last enzyme in the blood coagulation cascade. It is characterized by extensive genetic polymorphism defined by 4 common alleles, F13A*1A, 1B, 2A and 2B and a few rare variants, some responsible for severe coagulation deficiencies. In order to infer the evolutionary affinities between the common F13A alleles we have applied PCR techniques to study, in a Northern Portuguese sample, a short tandem repeat polymorphism located within the 5' untranslated region of the F13A gene. The analysis of the molecular heterogeneity within the F13A gene products revealed that the four biochemical variants shared very similar, truncated, distributions of STR alleles and showed no signs of predominant haplotypic associations. These findings seem to support both the inferences that intragenic recombination played an important role in the generation of molecular diversity within each of the four main F13A alleles and that all the four F13A alleles must be rather old. Molecular heterogeneity levels allowed the identification of 1B as the oldest F13A allelic state, and 2A as the most recently generated allele, but were not different enough to accurately track the divergence of alleles 1A and 2B. However, additional analysis of linkage disequilibrium patterns indicates that 1B-->2B-->1A-->2A is the most likely evolutionary order of appearance of F13A main protein alleles, confirming and extending a previous hypothetical model inferred from their molecular features. PMID- 10686500 TI - A note on a conditional-likelihood approach for family-based association studies of candidate genes. AB - The family-based association study design is a variation of the case-control study design, where unaffected family members instead of unrelated subjects are sampled as controls. This variation is useful in assessing the effects of candidate genes on disease, because it avoids false associations caused by admixture of populations. A complication of this design is that because of an inherited genotypic correlation among family members, the genotypic distributions between cases and relative controls may be distorted by the ascertainment criteria of families, which could involve not only cases and relative controls, but also other relatives. Analyzing such data naively may lead to biased estimates of relative risk. In this note, we will discuss the consistency of a conditional-likelihood approach. We show analytically that maximum conditional likelihood estimators are consistent for the true relative risks, if genotypes for family members are exchangeable under the sampling process, for example, sibling clusters. Besides being straightforward conceptually and computationally, this approach is robust to ascertainment bias and naturally accommodates genetic heterogeneity across families. PMID- 10686501 TI - Prenatal determination of a variable number of tandem repeats in intron 40 of the von Willebrand factor gene from maternal peripheral blood using the polymerase chain reaction. AB - To determine a fetal variable number of tandem repeats (VNTR) without using fetal tissues, we amplified the whole VNTR region in intron 40 of the von Willebrand factor gene from maternal peripheral blood by the polymerase chain reaction and then separated the amplified fragments by a nonhydratable polyacrylamide gel with an electrolyte gradient. After water elution from the gel, each preliminary product was amplified for the second time and then digested by Alu I restriction endonuclease. Then the VNTR was examined using a hydratable polyacrylamide gel with an electrolyte gradient. We successfully identified the fetal VNTR from 6 cases of 10 maternal peripheral blood samples and 2 cases of 6 maternal blood samples after parturition. This study might offer a theoretical basis for the noninvasive diagnosis of genetic disorders and identification of disputed paternity with common primers. PMID- 10686502 TI - Linkage analysis for diseases with variable age of onset. AB - We present a method for the multivariate linkage analysis of the age of onset of a disease. The approach allows the incorporation of covariates for the study of gene by environment interactions. It is applicable to general pedigrees. The likelihood of the data is expressed as a function of the number of alleles identical by descent at a marker, the censored ages of onset and disease status, and environmental exposures. In a simulation study, we compare the power to detect linkage under different sampling schemes for either a dominant or recessive trait when approximately 10% of individuals are gene carriers. The majority of the linkage information from a sample of randomly selected sib pairs was retained when the analyses were limited to sibships with one sibling having early-onset disease (<59 years old). Incorporating parental phenotypes could improve the power to detect the gene. When the sample consists of affected sib pairs (ASPs) having variable age of onset, the likelihood ratio (LR) test had higher power than the means (t(2)) test for detecting a locus with a large genetic relative risk (R(g) = 20). However, the power of the two tests was similar when ASPs are selected so that the proband has an early onset of disease. Lastly, the LR test had more power than the t(2) test to detect linkage in the presence of gene by environment interactions. PMID- 10686503 TI - Genetic and environmental factors contributing to the onset of allergic disorders. AB - Evidence has been accumulated to suggest that allergen-reactive Th2 cells play a triggering role in the activation and/or recruitment of IgE antibody-producing B cells, mast cells and eosinophils, the cellular triad involved in allergic inflammation. Recently, chemokines and chemokine receptors involved in such Th2 type response have been also defined. Th2 cells represent the polarized arm of the effector-specific responses that contribute to the protection against gastrointestinal nematodes and act as regulatory cells for chronic and/or excessive Th1-mediated responses. Th2 cells are generated from precursor naive Th cells when they encounter the specific antigen in an IL-4-containing microenvironment. The question of how these Th2 cells are selected in atopic patients is also unclear. Both the nature of the T cell receptor signalling provided by the allergen peptide ligand and a disregulation of IL-4 production likely concur to determine the Th2 profile of allergen-specific Th cells, but the genetic unbalanced IL-4 production is certainly overwhelming. Some gene products selectively expressed in Th2 cells or selectively controlling the expression of IL-4 have recently been described. These findings allow to suggest that the upregulation of genes controlling IL-4 expression and/or abnormalities of regulatory mechanisms of Th2 development and/or function may be responsible for Th2 responses against allergens in atopic people. The increasing prevalence of allergy in developed countries suggests that environmental factors acting either before or after birth also contribute to regulate the development of Th2 cells and/or their function. The reduction of infectious diseases in early life due to increasing vaccinations, antimicrobial treatments as well as changed lifestyle are certainly important in influencing the individual outcome in the Th response to ubiquitous allergens. Moreover, the recent evidence that bacterial DNA or oligodeoxynucleotides containing unmethylated 'CpG motifs' promote the development of Th1 cells via the production of immunomodulatory cytokines (namely IL-12, IL-18 and IFNs) by professional antigen-presenting cells confirms previous epidemiological data. The new insight into the pathophysiology of T cell responses in atopic diseases provides exciting opportunities for the development of novel immunotherapeutic strategies. PMID- 10686504 TI - Characterization of T cell subpopulations involved in the pathogenesis of asthma and allergic diseases. AB - Allergic asthma is a complex and heterogeneous disease which is characterized by intermittent reversible airway obstruction, chronic inflammation of the airways, bronchial hyperreactivity and an infiltration of lymphocytes and eosinophils into the airway submucosa. Animal models and clinical studies in humans have indicated an important role for T helper type 2 lymphocytes, producing IL-4, IL-5 and IL 13, in the pathogenesis of this disorder. However, although IL-4 and IL-13 have strong anti-inflammatory properties, the physiologic anti-inflammatory Th2 response does not seem to be operational in allergic asthma. Moreover, the induction of a Th1 response seems to aggravate, rather than ameliorate, its inflammatory character. This article will focus on the involvement of T lymphocyte subpopulations in the pathogenesis of allergic asthma and allergic diseases. In addition, a potential role of the subpopulation(s) of T regulatory cells in the induction and/or maintaince of the disease process will be discussed. PMID- 10686505 TI - Identification of sunflower seed IgE-binding proteins. AB - BACKGROUND: Sunflower seed can cause severe anaphylactic reactions in some susceptible individuals. It is conceivable that the 2S sunflower seed protein is an allergen based on its high degree of homology (34%) with the allergenic mature 2S albumin protein of the Brazil nut. The first step in determining the allergenicity of sunflower seed proteins is to identify IgE-binding proteins. METHODS: Sera from sunflower seed-sensitive individuals were evaluated by radioallergosorbent test (RAST), isoelectric focusing (IEF) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting with sunflower seed proteins. RESULTS: Positive RAST scores (>2) were observed in 3 individuals and immunoblotting demonstrated IgE-binding to 2-7 distinct proteins ranging in size from 10 to 50 kD. Two out of 3 sera recognized two proteins between 16 and 17 kD. The lower molecular weight protein (16 kD) approximates to the prepo region of the precursor methionine-rich 2S albumin protein found in sunflower seed (SFA 8/SSA). IEF followed by immunoblotting demonstrated several IgE-binding proteins, including two proteins with isoelectric points of 5.97 and 5.3, respectively, which are consistent with the mature and immature forms of the SFA-8/SSA region. CONCLUSIONS: Sunflower seed contains several IgE-binding proteins, including regions of the high-methionine 2S albumin SFA-8/SSA. PMID- 10686506 TI - The effect of antigen stimulation on alpha(4), beta(1) and beta(7) chain integrin expression and function in CD4+ cells. AB - BACKGROUND: The alpha(4) integrin, as alpha(4)beta(1) (VLA-4) or alpha(4)beta(7), is critical for T cell migration and proliferation, although its functional modulation remains poorly understood. We hypothesized that increased receptor density, based on new receptor chain synthesis, was one such mechanism. We examined the surface receptor density of the alpha(4) and beta(1) chains on CD4+CD45RO+ cells, and the mRNA expression of these and the beta(7) chain in response to allergen and nonallergen antigen stimulation. METHODS: Flow cytometric analyses for CD49d, CD29, and CD45RO were performed on T cell lines specific for timothy, tetanus, and Candida from atopic and nonatopic donors. RNA was extracted from cells sorted to select CD4+/CD49d-positive cells before and after stimulation. Equivalent amounts of cDNA for beta-actin, alpha(4), beta(1) and beta(7) were used in PCR, and the products were quantified using phosphoimaging. RESULTS: CD49d expression is heterogeneous on T cell lines and is upregulated by antigen stimulation on CD4+ T cells. The surface expression on CD4+CD45RO+ timothy allergen or tetanus toxoid T cell lines is at least double that found on CD45RO- cells. Antigen stimulation upregulated CD49d expression on the CD4+CD45RO+ subpopulation of both cell lines although it was not as significant as in the case of all CD4+ T cells. CD29 surface expression behaves similarly. Candida had no effect on CD49d or CD29. Messenger RNA expression for the alpha(4) chain (CD49d) is significantly upregulated 48 h following the addition of timothy or tetanus. beta(7) chain expression also rises significantly on both cell lines. beta(1) chain expression increases, but not significantly. CONCLUSIONS: The surface expression of the CD49d is heterogeneous and much higher on CD4+CD45RO+ cells than on CD4+RO- T cells. The CD49d integrin chain on CD4+ T cells is upregulated following antigen exposure. However, the CD4+CD45RO+ subpopulation is only partially responsible for this increase suggesting other T cells to have this receptor expression upregulated. CD29 expression behaves similarly. Messenger RNA expression increases coordinately for alpha(4), beta(7), and not significantly for beta(1) in these cells. These observations provide a potential mechanism for the selective accumulation of T cells at sites of inflammation, and suggest an important point of intervention for allergic and inflammatory disease. PMID- 10686507 TI - Effect of Ca(2+) ATPase inhibitors on MCP-1 release from bone marrow-derived mast cells and the involvement of p38 MAP kinase activation. AB - The effect of two Ca(2+) ATPase inhibitors, cyclopiazonic acid (CPA) and 2,5-di (tert-butyl)-1,4-hydroquinone (DTBHQ), on the release of MCP-1 from bone marrow derived mast cells (BMMCs) were investigated. CPA and DTBHQ increased the intracellular free Ca(2+) concentration ([Ca(2+)](i)) and induced MCP-1 release in a dose-dependent manner. These Ca(2+) ATPase inhibitors induced MCP-1 release in the absence of phorbol ester, in contrast to their induction of TNF-alpha. MCP 1 release reached a maximum at 6-9 h. It was inhibited by treatment with actinomycin D, the immunosuppressant cyclosporin A, and the cytosolic Ca(2+) chelator BAPTA-AM. Furthermore, RT-PCR showed a time-dependent increase of MCP-1 mRNA. Thus MCP-1 release seems to depend on Ca(2+)-dependent transcriptional activation. MCP-1 release was dose-dependently inhibited by the p38 MAP kinase inhibitor SB202190, but not by the p44/42 MAP kinase inhibitor PD98059. Therefore, transcriptional activation of MCP-1 production and its release seem to be dependent on the nuclear factor of activated T cells and p38 MAP kinase activation. This is the first report to show the regulation of MCP-1 production in BMMCs. PMID- 10686508 TI - Sodium salicylate-induced apoptosis of human peripheral blood eosinophils is independent of the activation of c-Jun N-terminal kinase and p38 mitogen activated protein kinase. AB - BACKGROUND: It has been shown that the inhibition of eosinophilic apoptosis is an important mechanism for the development of blood and tissue eosinophilia in allergic diseases. Considerable attention has recently been focused on the role played by different intracellular kinase cascades in the control of apoptosis. In the present study, we investigated the effect of sodium salicylate (NaSal), a nonsteroidal anti-inflammatory drug, on mitogen-activated protein kinases (MAPK) and apoptosis of human eosinophils. METHODS: Human blood eosinophils were purified from buffy coat. NaSal-induced apoptosis of eosinophils was assessed by morphological changes and Annexin-V binding assay. Changes of MAPK activity upon treatment with NaSal were measured by kinase activity assay and Western blot. RESULTS: NaSal could induce apoptosis of human blood eosinophils in a dose- and time-dependent manner. It could also activate c-Jun N-terminal kinase (JNK) and p38 MAPK but not extracellular signal-regulated protein kinase (ERK) activity within 1 h. Pretreatment of eosinophils with p38 MAPK and JNK anti-sense (AS) phosphorothioate oligodeoxynucleotides (ODN) or specific p38 MAPK inhibitor SB 203580 did not have any significant effect on NaSal-induced apoptosis. However, ERK AS ODNs could trigger the apoptosis of normal eosinophils. CONCLUSION: There is no direct relationship between the activation of JNK and p38 MAPK pathways and NaSal-induced apoptosis in human peripheral blood eosinophils. PMID- 10686509 TI - Neurokinin-1 receptor antagonist inhibits short-term sulfuric-acid-induced airway hyperresponsiveness in sensitized guinea pigs. AB - BACKGROUND: Tachykinins are involved in the development of bronchial inflammation and airway hyperresponsiveness (AHR); however, the role of the neurokinin-1 (NK(1)) receptor in acid-aerosol-induced bronchial impairment in asthmatic patients remains controversial. METHODS: To investigate the effects on the NK(1) receptor antagonist FK888 the neurokinin-2 (NK(2)) receptor antagonist SR48968 on sulfuric-acid (H(2)SO(4))-induced AHR in guinea pigs, specific airways resistance (sRaw) and airways responsiveness to methacholine (MCh) were measured before and after 6 h of exposure to H(2)SO(4) aerosol (pH 1.7, 82 mg/m(3)) in ovalbumin sensitized guinea pigs. RESULTS: Airway responsiveness to MCh significantly increased (p<0. 05) after the exposure, however sRaw did not. Treatment with FK888 significantly inhibited (p<0.05) H(2)SO(4)-induced AHR in a dose-dependent manner, as did SR48968. CONCLUSIONS: These results suggest that not only NK(2) but also NK(1) receptors might have important roles in the development of acid aerosol-induced AHR. PMID- 10686510 TI - Sequential development of airway hyperresponsiveness and acute airway obstruction in a mouse model of allergic inflammation. AB - BACKGROUND: Mouse models have been established mirroring key features of human bronchial asthma including airway hyperresponsiveness (AHR). Acute airway obstruction in response to an allergen challenge, however, remains to be demonstrated in these models. OBJECTIVE: A mouse model of allergic lung inflammation was employed to analyze the development of specific (allergen induced) and nonspecific (methacholine-induced) airway obstruction. METHODS: Mice were sensitized to ovalbumin (OVA) and challenged with OVA aerosol twice each week during four weeks. Changes in lung functions were determined by noninvasive head-out body plethysmography. The development of acute airway obstruction after OVA challenge and AHR after methacholine aerosol application were assessed by a decrease in the mid-expiratory flow rate (EF(50)). RESULTS: Two airway challenges were sufficient to induce AHR (5.7 vs. 15 mg/ml methacholine). Further OVA challenges reduced the baseline EF(50) from 1.85 to 1.20 ml/s (4th week) and induced acute airway obstruction. The OVA-induced obstruction was maximal in the 4th week (EF(50) = 0.91 ml/s). CONCLUSION: The development of acute airway obstruction in allergen-sensitized mice was demonstrated by means of head-out body plethysmography. In our model, AHR was observed before the development of airway obstruction. PMID- 10686511 TI - Characterization of tissue outgrowth developed in vitro in patients with rheumatoid arthritis: involvement of T cells in the development of tissue outgrowth. AB - BACKGROUND: The aim of this study was to analyze cellular and cytokine interactions governing the development of synovial tissue outgrowth in patients with rheumatoid arthritis (RA). METHODS: A single-cell suspension of dissociated synovial tissues of RA patients was cultured for a long period to develop tissue outgrowth. The resulting tissue outgrowth was characterized by immunohistochemical staining and ELISA. RESULTS: The tissue outgrowth developed in vitro included various cell types, such as macrophage-like synovial cells, fibroblast-like synovial cells and lymphocytes. Even after prolonged cultivation, synovial cells devoid of infiltrating T lymphocytes did not form tissue outgrowth. The outgrowth contained CD3+ cells, LeuM3 (CD14)+ cells and HLA-DR+ cells. The T cells expressed lymphocyte function-associated antigen (LFA)-1 and CD2, and the synovial cells expressed intracellular adhesion molecule (ICAM)-1 and LFA-3, suggesting possible interactions via LFA-1/ICAM-1 and CD2/LFA-3. Production of T-cell derived IFN-gamma and IL-17 and synovial-cell-derived fibroblast growth factor (FGF)-1 and IL-15 was confirmed in the tissue outgrowth as well as in RA synovial tissue. These cell types stimulate each other by secreting cytokines, leading to the secretion of proinflammatory cytokines and matrix metalloproteinase (MMP)-1 by the tissue outgrowth and proliferation of both lymphocytes and synovial cells. CONCLUSION: This study emphasizes the importance of cellular interactions between T cells and synovial cells, via adhesion molecules and the secretion of cytokines with stimulatory activity towards other cell types, for the hyperactivity of RA synovial cells. PMID- 10686512 TI - Immunogenicity of neutralizing epitopes on multiple-epitope vaccines against HIV 1. AB - Based on our hypothesis that epitope vaccine may be a new strategy to induce high levels of neutralization antibodies against HIV-1, we prepared multiple-epitope vaccines using three neutralizing epitopes (GPGRAFY, RILAVERYLKD and ELDKWA) of HIV-1 gp160, and characterized their immunogenicity. Peptide 1 [C-G-(ELDKWA GPGRAFY)(2)-K] and peptide 2 (CG-GPGRAFY-ELDKWA-G-RILAVERYLKD) were synthesized and conjugated with carrier protein bovine serum albumin (BSA). After vaccination antibody responses to these immunogens were induced and evaluated by ELISA. The C G-(ELDKWA-GPGRAFY)(2)-K-BSA (BSA: carrier protein) multiple-epitope vaccine induced a strong antibody response to the C-G-(ELDKWA-GPGRAFY)(2)-K peptide (antibody titer: 1:25,600) and C-(ELDKWAG)(4) peptide (antibody titer: 1:12,800), but a weak antibody response to the C-(GPCGRAFY)(4) peptide. The CG-GPGRAFY ELDKWA-G-RILAVERYLKD-K-BSA (BSA: carrier protein) multiple-epitope vaccine also induced strong antibody response to the CG-GPGRAFY-ELDKWA-G-RILAVERYLKD-K peptide (antibody titer: 1:25, 600) and C-(ELLDKWAG)(4) peptide (antibody titer: 1:6,400), a very strong response to C-(RIVALVERYLKD-G)(2)-K peptide (dilution: 1:102, 400), and a very weak response to the C-(GPGRAFY)(4) peptide (dilution: 1:400) in mice. Both antisera induced by both multiple-epitope vaccines interacted with the recombinant soluble gp41 (rgp41), but did not bind two control peptides. In comparison with both epitope vaccines, the rgp160 subunit vaccine could induce weak epitope-specific antibody response to these three epitopes on the three epitope peptides and V3, N-domain and C-domain peptides (dilution: 1:400-1:1,600). These results indicate that both multiple-epitope vaccines could induce high levels of antibodies to both neutralizing epitopes RILAVERYLKD and ELDKWA, while the GPGRAFY epitope on both vaccines appeared to have weak immunogenicity. Both multiple-epitope vaccines showed significant potency on inducing high levels of epitope-specific neutralization antibodies in comparison with rgp160 subunit vaccine. PMID- 10686513 TI - A simple advice for the prevention of pollen-induced allergic rhinitis. PMID- 10686514 TI - Castration of sexually immature rats affects sympathetic innervation of the adult thymus. AB - It has been hypothesized that maturational processes within the hypothalamo pituitary-gonadal (HPG) axis and thymus are reciprocally regulated via neural pathways. To test this hypothesis, in the thymi of adult rats orchidectomized (ORX) at age of 1 (ORX-1), 7 (ORX-7) and 30 days (ORX-30): (i) noradrenaline (NA), dopamine (DA) and serotonin (5-HT) contents and acetylcholinesterase (AChE) activity were measured and (ii) the distribution of monoamine- and AChE containing nerves and cells was examined by a sucrose phosphate glyoxylic acid (SPG) method and enzyme histochemistry, respectively. In all groups of ORX rats, the thymus weight was significantly increased over that in sham-ORX control rats. In the ORX-1 rats, the increase in the thymus weight was accompanied by a proportional increase in the content of both catecholamines and 5-HT; consequently the concentration of each of them remained unaltered. In these animals, the density of both SPG-stained thymus nerve fibers and cells also remained unchanged. In the ORX-7 rats, the rise in the thymus weight was followed by a proportional increase in the content of all monoamines, except for NA which was reduced. Therefore, in these rats neither the thymus concentrations of DA nor that of 5-HT differed from controls, while the concentration of NA was significantly decreased. The reduction in both NA content and concentration reflected a diminished density of SPG-positive nerve profiles. In the ORX-30 rats, the increase in thymus weight was neither paralleled by a proportional increase in the DA content nor in 5-HT, while the content of NA was decreased. Thus, in their thymi the concentration of both NA and DA, as well as that of 5 HT, were significantly reduced. In parallel with these changes, a decreased density of thymic SPG-positive nerve fibers and cells was found. In all ORX rats, the pattern of intrathymic distribution of SPG-positive fibers and cells remained unchanged. Orchidectomy affected neither the activity of AChE (expressed per gram of tissue) nor the density of AChE-positive nerves and cells in the thymus. As the changes in the density of adrenergic nerve fibers in the thymus from ORX rats were not followed by similar alterations in the density of AChE-containing nerve fibers, it does not seem likely that NA and AChE are colocalized in the thymus nerve fibers. The results also suggest that there is a critical period during ontogenesis when changes within the HPG axis evoked by orchidectomy can affect the sympathetic nerve input to the rat thymus and therefore, most likely, development and function of the organ. PMID- 10686515 TI - Suppression of lymphocyte mitogenesis by tamoxifen: studies on protein kinase C, calmodulin and calcium. AB - The effect of tamoxifen (TX; 1.0 microM) on the mitogenic response of rat lymphocytes was compared with the effect of drugs that are known to act on protein kinase C (PKC), calmodulin (CM), and calcium (Ca(2+)). The calcium ionophore A23187 (0.2 microM) was mitogenic on its own which was not influenced by TX. The agents modulating PKC or CM (phorbol-myristate-13-acetate; R24571, chlorpromazine) influenced mitogenesis differently than did TX. General inhibition of lymphocyte proliferation was seen with the Ca(2+) antagonist agents (EGTA, TMB-8) as with TX. The antiproliferative effect of TX was partially reversed by the increase of Ca(2+) in the culture medium when T cell mitogens were used, but not in the case of lipid A, a B lymphocyte mitogen. However, the concanavalin A-induced Ca(2+) influx was further elevated by TX which differed from the effect of the Ca(2+) channel-blocking agent verapamil. The results suggest that TX resets the threshold stimulus necessary for mitogenesis and is completely reversible. PMID- 10686516 TI - Effect of interleukin-6 and tumor necrosis factor-alpha on GABA release from mediobasal hypothalamus and posterior pituitary. AB - The release of cytokines during infection, inflammation and stress induces brain mediated responses, including alterations of neuroendocrine functions. We examined the effect of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha) on release of gamma-aminobutyric acid (GABA) from mediobasal hypothalamic (MBH) explants and posterior pituitaries (PP) of male rats. IL-6 (10 ng/ml) did not modify basal GABA release from MBH and PP, but significantly increased GABA release under depolarizing conditions (40 mM K(+)). This effect was abolished by incubation of the tissue with indomethacin, an inhibitor of cyclooxygenase activity, indicating that prostaglandins could mediate the stimulation of GABA release induced by IL-6. On the contrary, TNF-alpha (50 ng/ml) significantly decreased K(+)-evoked GABA release from both MBH and PP. This inhibitory effect was not modified by indomethacin. Neither IL-6 nor TNF-alpha affected nitric oxide synthesis, as measured by [(14)C]citrulline production. The current results indicate that IL-6 stimulates GABA release from both hypothalamus and posterior pituitary by a mechanism mediated by prostaglandins. On the contrary, TNF-alpha inhibits GABA release from both tissues. These results suggest the possibility that GABAergic activity in the hypothalamic-pituitary axis could be involved in neuroendocrine responses to cytokines. PMID- 10686517 TI - Time-dependent effect of cyclosporine on mitogenic responses and lymphocyte subset populations in rat spleen and submaxillary lymph nodes. AB - Lipopolysaccharide (LPS)- and concanavalin A (ConA)-induced proliferation and T lymphocyte subsets were measured in spleen and submaxillary lymph nodes of male rats injected with cyclosporine (5 mg/kg s.c. for 5 days, at 12.00 or 24.00 h; animals kept under light from 08.00 to 20.00 h daily). One hour before the third injection, Freund's complete adjuvant or its vehicle was administered. A suppressive effect of cyclosporine on the mitogenic action of LPS was seen in the spleen of rats injected with cyclosporine at noon whereas the effect of ConA remained unaffected. CD4+, CD8+ and CD4+-CD8+ cells decreased in spleen and lymph nodes of Freund's adjuvant-injected rats only with cyclosporine given at noon. The results further support occurrence of time-of-day-dependent effects of cyclosporine on lymphocyte subsets and proliferation. PMID- 10686518 TI - Time of day-dependent effects of thyroliberin and thyrotropin on thymocyte proliferation in rats. AB - Our earlier studies have shown that thyroliberin (TRH) as well as thyrotropin (TSH) enhanced thymus cell proliferation. The aim of the present study was to investigate whether the effects of TRH and TSH on thymocyte proliferation depend on time of day. A single subcutaneous injection of TRH (25 microgram/animal) or TSH (3 IU/animal) was made at 9:00 h (1 h after light onset) or at 18:00 h (10 h after light onset) in 3-month-old male Wistar rats. The animals were killed 24 h later. The proliferation of thymocytes was assessed by incorporation of bromodeoxyuridine into cell nuclei. Thymocyte proliferation was significantly increased by TSH administration at 9:00 h, whereas treatment given at 18:00 h was ineffective. TRH enhanced the proliferation when injected at 9:00 h, but had an inhibitory effect when administered at 18:00 h. These data indicate that the effects of TRH and TSH on thymocyte proliferation are dependent on time of day. PMID- 10686519 TI - Metabolic, neuroendocrine and immune functions in basal conditions and during the acute-phase response to endotoxic shock in undernourished rats. AB - Chronic malnutrition is one of the most important causes of several metabolic, immune and neuroendocrine dysfunctions. The aim of the present study was to determine the influence of chronic food restriction on basal neuroendocrine, immune and adipocyte functions and during the acute-phase response to endotoxic shock in female rats. The effect of refeeding of undernourished rats on the above mentioned functions was also investigated. For these purposes, plasma total protein, glucose, triglycerides, ACTH, corticosterone, tumor necrosis factor alpha (TNF) and leptin (LEP) levels were determined in basal condition and 2 h after endotoxin (LPS; 180 microgram/kg body weight, i.p.) administration in 3 different groups: (1) well-nourished (WN) controls; (2) undernourished (UN) rats as a consequence of chronic food restriction, and (3) UN rats re-fed to restoration of their body weights in the WN rat range. The results indicate that UN rats, in comparison with WN controls, developed an arrest in body weight gain as well as in basal hypoglycemia, hypotriglyceridemia, hypoleptinemia, hypercorticosteronemia and enhanced adrenal glucocorticoid content; however, no changes in basal total protein, ACTH and TNF plasma levels and in anterior pituitary ACTH concentrations were found. When endotoxic shock was induced, the LPS-induced hypoglycemia developed in WN rats was abolished in UN animals, and both ACTH and TNF plasma concentrations after endotoxin, albeit significantly (p < 0.05) higher than the respective basal values, were significantly (p < 0.05) lower in UN than in WN control rats. Despite the high basal plasma corticosterone concentration in UN vs. WN rats, the LPS-induced glucocorticoid release was similar in WN and UN rats. Additionally, LPS treatment did not modify basal plasma LEP levels, regardless of the group. Interestingly, UN rats fed ad libitum for 15 days restored their body weight to WN rat range values, and the various metabolic dysfunctions seen in UN rats in both basal and post-LPS conditions were fully normalized. Our results clearly indicate that chronic undernutrition not only affects, as earlier described, reproductive function but also metabolic, neuroendocrine, immune and adipocyte functions, and that the effects induced by undernutrition can be fully reversed after recovery of normal body weight. The present study strongly supports the involvement of the metabolic status in the effectiveness of the defense mechanisms developed in patients in inflammatory stress conditions. PMID- 10686520 TI - Caffeic acid phenethyl ester induces leukocyte apoptosis, modulates nuclear factor-kappa B and suppresses acute inflammation. AB - Nuclear factor kappa-B (NF-kappaB) is a heterodimeric transcription factor with a pivotal role in orchestrating immune and inflammatory processes. Inflammatory cytokines and prostanoids activate NF-kappaB, which, in turn, stimulates expression of cytokines, proteases, adhesion molecules and other inflammatory mediators. Caffeic acid phenethyl ester (CAPE) is a compound that modulates nuclear binding of the NF-kappaB p65 subunit (RelA). To determine whether CAPE decreases the viability of cells participating in host defense, we first tested its in vitro effect on a glucocorticoid-sensitive and -resistant cell line of lymphoid origin. CAPE induced apoptotic cell death in a dose-dependent fashion and to a similar extent in both cell lines. Furthermore, a low concentration of CAPE decreased the LD(50) of dexamethasone by 3- to 5-fold. Since therapeutic induction of apoptosis of activated inflammatory cells holds the attraction of destroying effector cells safely without secondary tissue damage, we examined the effects of CAPE in a rat model of carrageenin-induced subcutaneous inflammation. Local administration of CAPE resulted in increased leukocyte apoptosis and marked reduction in exudate leukocyte, neutrophil and monocyte concentrations at the inflammatory site. CAPE decreased expression of cytosolic IkappaBalpha and increased nuclear translocation of p65. These findings may suggest that novel anti-inflammatory therapies can be based upon activation of NF-kappaB-mediated transcription of genes curbing the inflammatory response and that CAPE or its analogs hold therapeutic promise. PMID- 10686521 TI - Conditioned alterations of specific blood leukocyte subsets are reconditionable. AB - Behavioral conditioning has the ability to produce changes in immune function. However, it is unknown whether conditioned changes of immune function can be recalled on multiple occasions. To address this issue we paired a novel saccharin drinking solution with intraperitoneal cyclosporin A (CsA) injection in rats. Saccharin re-presentation produced a reduction in splenocyte proliferation that mirrored the effect of CsA. Such functional changes were paralleled by a significant conditioned leukopenia in peripheral blood, which opposed the leukocytosis induced by CsA. Using the conditioned leukopenia in blood as a 'diagnostic window' of conditioned immunosuppression, the maintenance of CsA induced changes was investigated by examining blood samples collected repeatedly. Experiments on the same group of animals over a period of 1 year showed that the conditioned leukopenia was reproducible on multiple occasions by reimplementing either the whole conditioning paradigm or reexposure to the saccharin solution only. These results demonstrate that behaviorally conditioned alterations of immune parameters are maintained in subsequent trials, indicating the potential clinical feasibility of behavioral conditioning procedures. PMID- 10686522 TI - CRH deficiency impairs but does not block pituitary-adrenal responses to diverse stressors. AB - We have previously observed significant, albeit decreased, corticosterone responses to restraint stress in corticotropin releasing hormone (CRH)-deficient (knockout, CRH KO) mice. Because different stressors have been shown to engage different populations of hypophysiotropic neurons, we have used hypoglycemia and hypovolemia to test whether CRH-independent pituitary-adrenal activation is evoked by stimuli other than restraint. Insulin injection in fasted CRH KO mice elicited increases in corticosterone that were markedly lower than those in wild type but marginally significant relative to corresponding KO controls. Consistent with impaired adrenocortical function, hypoglycemia-induced epinephrine secretion was reduced in female CRH KO mice. Hypovolemia produced by retro-orbital bleeding also significantly elevated corticosterone in CRH KO mice. In contrast to significant stress-induced increases in corticotropin (ACTH) in wild-type mice, those in CRH KO mice were slight, transient and difficult to detect without frequent sampling. Restraint-induced interleukin-6 (IL-6) levels were similar between wild-type and CRH KO mice, arguing against compensatory changes in IL-6 responses to restraint due to CRH deficiency. CRH infusion enhanced adrenocortical responses to restraint independently of effects on basal corticosterone levels, suggesting that pituitary-adrenal activity is augmented by factors besides CRH during stress. We conclude that although stress-induced pituitary-adrenal activity does not require acute increases in CRH, CRH is required to support the normal amplitude of adrenocortical axis responsiveness to other endocrine or neural factors during stress. PMID- 10686523 TI - Potentiated response of corticotropin (ACTH) to repeated moderate hemorrhage requires amygdalar neuronal processing. AB - We examined the role of the amygdala in the potentiation of the corticotropin (ACTH) response to a 10 mg/kg hemorrhage by a 1-hour episode of equivalent hypovolemia done 24 h earlier. Unanesthetized rats were studied on the fourth (D1) and fifth (D2) day after chronic implantation of arterial and venous catheters. Immunocytochemistry for Fos protein indicated that neurons in the central and medial nuclei of the caudal amygdala were activated by hemorrhage. We then tested the effect of excitotoxic destruction of the neurons in these areas by bilateral injections of ibotenic acid 10 days prior to catheter placement. In rats that were hemorrhaged on both D1 and D2, the responses of ACTH and corticosterone increased significantly from the first (H1) to the second hemorrhage (H2) in a control group injected with saline (p < 0.05) and in lesioned groups without bilateral damage of the Fos-responsive areas (p < 0.01). In the group with bilateral damage to these sites, the responses to H1 and H2 did not differ. Additional rats had H1 on D2 to control for the long-term effects of the chronic cannulation. The responses of ACTH to H1 on either D1 or D2 did not differ between the saline-injected controls and any of the lesioned groups. In contrast, the response of ACTH to H2 on D2 in rats with bilateral damage of the caudal amygdala was not significant and was less than the response of ACTH to H2 in both rats with unilateral damage of this area (p < 0.05) and those injected with saline (p < 0.05). We conclude that bilateral neuronal processing within the caudal amygdala is required for the potentiation of the response of ACTH to H2 by H1. PMID- 10686524 TI - Monensin and hypo-osmolar medium cause calcium-independent beta-endorphin secretion from melanotropes. AB - Monensin has been shown to cause nonexocytotic release of catecholamines from adrenal medullary and PC12 cells. We examined the effect of monensin on peptide secretion with cultured melanotropes from the rat pituitary as a model. 1 microM monensin caused an immediate, transient increase in beta-endorphin secretion. The effect was still seen in a calcium-free medium, but was totally abolished in a sodium-free medium. Intracellular calcium concentration was measured with Fura 2: no increase was observed during monensin stimulation. Hypo-osmolar medium mimicked the effect of monensin, causing a 12-fold transient increase in beta endorphin secretion. This effect was not abolished in either calcium-free or sodium-free medium. No increase in the number of exocytotic figures captured by tannic acid incubation was observed during 5 min of incubation with 1 microM monensin or hypo-somolar medium. We thus show that monensin causes beta-endorphin secretion from the melanotrope and that this effect is due to sodium influx and resultant cell swelling. The calcium independency and lack of increase of exocytotic figures suggest that swelling-induced secretion is nonexocytotic, possibly via transient exocytotic pore opening. PMID- 10686525 TI - Hyperinnervation during adrenal regeneration influences the rate of functional recovery. AB - The rat adrenal cortex has the uncommon ability to demonstrate morphological and functional regeneration after injury-induced loss of cortical tissue. Peripheral nerves are involved in tissue regeneration and healing after injury, implying that nerves may also be involved in modulating the regeneration of the adrenal cortex. Studies were initiated to assess changes in adrenal innervation during cortical tissue regeneration subsequent to adrenal enucleation. Innervation of regenerating adrenals was assessed from 3 to 62 days postenucleation by immunohistofluorescent detection of neuronal markers for primary afferent, preganglionic sympathetic, and postganglionic sympathetic fibers. The regenerating adrenal contained few nerves at 3 days postenucleation, but became differentially innervated, with extensive innervation by nerve fibers positive for calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH), neuropeptide Y (NPY), and neuronal nitric oxide synthase (nNOS). In contrast, there was only minimal innervation by nerve fibers positive for vasoactive intestinal peptide. By 14 days postenucleation, the CGRP-, TH-, and NPY-positive innervation included areas of hyperinnervation in the capsule, cortex, and central inflammatory site of the regenerating gland. In addition, many chromaffin cells were present at all time points postenucleation. Quantification of the regenerating gland content of CGRP, norepinephrine, epinephrine, and nNOS verified the immunohistofluorescent observations. The period of extensive innervation correlated temporally with the time (3-30 days) during which the regenerating glands recovered steroidogenic function. Moreover, splanchnic nerve transection at the time of adrenal enucleation decreased the innervation by CGRP positive and vesicular acetylcholine transporter-positive fibers and delayed regeneration. These results support the hypothesis that adrenal innervation modulates tissue regeneration and functional recovery of the enucleated adrenal gland. PMID- 10686526 TI - Far upstream sequences regulate the human prolactin promoter transcription. AB - The human prolactin gene is mainly expressed in pituitary lactotrope cells, but transcription from an alternative, far upstream promoter was detected in lymphoid, placental and mammary cells. We describe the transcriptional activity in rat pituitary cells of the complete region separating the two promoters, using transient transfection experiments. A far upstream activating region was only functional in combination with the prolactin promoter. DNaseI protection experiments revealed, in addition to binding sites for the pituitary-specific factor Pit-1, sites (e.g. SD1) for several ubiquitous factors and one lymphoid specific factor (SD4). A single copy of the ubiquitous site SD1 or the lymphoid specific site SD4 was unable to activate transcription of a heterologous promoter in pituitary cells. However, SD1 activated transcription in nonpituitary cells and SD4 was functional specifically in lymphoid cells. Five copies of a distal site (D8) activated transcription in each cell type tested. Gel retardation experiments show that this site binds the specific factor C/EBP in liver and a distinct factor in other cell types. Our results suggest that different elements within this large region direct specific expression from each promoter via a complex interplay between cell-specific and ubiquitous transcription factors. PMID- 10686527 TI - Pulsatile gonadotropin-releasing hormone (GnRH) secretion is an inherent function of GnRH neurons, as revealed by the culture of medial olfactory placode obtained from embryonic rats. AB - To determine whether gonadotropin-releasing hormone (GnRH) neurons in culture without the hypothalamus secrete GnRH in a pulsatile fashion, the nasal placode (NAP) was obtained at day 13.5 of gestation and cultured by a roller tube method. If the GnRH release occurs in a pulsatile fashion, it can be said that the pulse generator of GnRH exists inherently in each cell or community of cells in the culture. The concentration of GnRH in the NAP culture medium collected at 8-min intervals for 160 min after 2- to 4-week cultures showed that GnRH release occurred in a pulsatile fashion with a mean interpulse interval of 29.8 +/- 2.3 min (n = 9). When the NAP was cultured with tissues of the forebrain vesicle (n = 3) or the hypothalamus (n = 4), GnRH was also released in a pulsatile fashion with similar intervals (27.3 +/- 1.0 min for the NAP+forebrain vesicle culture and 36.0 +/- 6.3 min for the NAP+hypothalamus culture) as those in cultures without brain tissues. It is concluded that pulsatile GnRH release is an inherent function of GnRH neurons. PMID- 10686528 TI - Influence of different serotonin receptor subtypes on growth hormone secretion. AB - The role of serotonin (5-HT) in the regulation of growth hormone (GH) secretion remains unclear due to the existence of many different receptors that mediate the 5-HT actions, and the lack of suitable specific agonist and antagonist drugs. In the present work we have taken advantage of the recent development of new selective 5-HT drugs in order to clarify the role played by different 5-HT receptor types and subtypes on GH secretion. The experiments were carried out on beagle dogs. GH-releasing hormone (GHRH) increased basal canine GH (cGH) levels from 0.8 +/- 0.2 to 8.8 +/- 1.7 microg/l at 15 min. Administration of 5-HT(1D) receptor agonist sumatriptan (SUM) induced a cGH peak at 30 min of 12.9 +/- 2.7 microg/l. The combined administration of GHRH plus SUM strikingly potentiated cGH release with a peak of 36.9 +/- 6 microg/l at 30 min (p < 0.05). Pretreatment with the muscarinic receptor antagonist atropine completely abolished the cGH response to SUM, while the cholinergic agonist pyridostigmine (PYR) did not modify this response (15.3 +/- 5 microg/l PYR plus SUM vs. SUM alone 12.9 +/- 2. 7 microg/l). On the other hand, administration of drugs with activity at 5 HT(2A/C) receptors showed a stimulatory role for the 5-HT(2C) receptor subtype, since LY-53857 (antagonist 5-HT(2A/C)) and DOI agonist (5-HT(2A/C)) both modified the GH response stimulated by GHRH (AUC 88.5 +/- 30.4 and 400 +/- 64.6 vs. 267.3 +/- 52.6 respectively), while ketanserin (antagonist 5-HT(2A)) did not modify this response. The 5-HT(3) antagonist ICS-205-930 failed to modify either basal or GHRH induced GH responses. In conclusion, our data show that 5-HT(1D) receptors play a stimulatory role on GH secretion in the dog, possibly by acting through a decrease in hypothalamic somatostatin release. Similarly, the 5-HT(2C) receptor subtypes also appear to play a stimulatory role. However, 5-HT(2A) and 5 HT(3) receptors do not appear to be involved in the control of basal and GHRH induced GH secretion. PMID- 10686529 TI - How is Creutzfeldt-Jakob disease acquired? AB - Creutzfeldt-Jakob disease (CJD) is one of several related disorders collectively called prion diseases. These disorders affect man and animals and are now known to be caused by the abnormal configuration of a naturally occurring protein, PrP(c). By mechanisms still not well understood, this natural protein is converted into a pathologic variant, PrP(sc). The disease is 'acquired' spontaneously perhaps by posttranslational conversion of a PrP(c) into a PrP(sc) population. This sporadic form of CJD has been reported worldwide with a frequency of 1/million. Other modes of acquisition include the following: ingestion of brain tissue from deceased victims through ritual cannibalism at burial ceremonies formerly (and no longer) practiced by New Guinea Highlanders; iatrogenically, through corneal transplants from infected donors, inoculation of human growth hormone and gonadotropin prepared from infected human pituitary glands; from inadequately sterilized depth electrodes introduced neurosurgically into the brain during workups of patients with epilepsy, and applications of infected dura mater in neurosurgical procedures. Most recently, an infected bovine source (bovine spongiform encephalopathy) has been implicated and produces a new variant of CJD. Clusters of CJD in families in some populations have been recognized which are inherited in Mendelian fashion. These clusters are related to mutations of the PRNP gene in specific codons (e.g. codon 200). Homozygosity for these mutations increases the chances of manifesting the disease. Other potential methods of acquisition, such as by blood transfusion, surgical sutures, tonometers, consumption of hog brain or other organs and tissue, remain unproven. PMID- 10686530 TI - Creutzfeldt-Jakob disease in a recipient of a dura mater graft processed in the US: cause or coincidence? AB - Iatrogenic Creutzfeldt-Jakob disease (CJD) has never been reported among recipients of dura mater grafts processed in the US. We recently investigated a report of such a case in a 72-year-old man with a typical clinical presentation of CJD. We found no evidence of CJD in either the 34-year-old donor or in other, proximal patients undergoing craniotomies. Although the graft may have caused the illness, sporadic CJD is a more likely explanation, with the graft being coincidental. PMID- 10686531 TI - Regional and temporal variation in the incidence of multiple sclerosis in Finland 1979-1993. AB - Previous surveys in Finland from the 1960s have documented an uneven geographic distribution of multiple sclerosis (MS). In the present study, the incidence of MS was studied during 1979-1993 in the western Vaasa and Seinajoki regions and in southern Uusimaa. The overall difference between the western and southern regions persisted; 8.7 per 100,000 in the western, and 5.1 per 100,000 in the southern region. The incidence of 11.6 per 100,000 in Seinajoki was more than twofold greater than the 5.2 per 100,000 incidence found in neighboring Vaasa. An increasing incidence trend was observed for men in Seinajoki, and a decrease for both sexes in Vaasa, while in Uusimaa the incidence remained stable for both sexes. The different incidence trends could not be readily explained by differences in case ascertainment but suggest the effect of environmental factors that have modulated the incidence of MS during the 15-year study period. PMID- 10686532 TI - Relationship of family history scores for stroke and hypertension to quantitative measures of white-matter hyperintensities and stroke volume in elderly males. AB - White-matter hyperintensities (WMHI) are frequently associated with cerebrovascular risk factors in the elderly, particularly hypertension, and have been interpreted as a subclinical form of ischemic brain damage. WMHI, clinical stroke and blood pressures show significant genetic influences. The objective of this study was to determine whether a relationship exists between family history of stroke and/or hypertension in first degree relatives and WMHI in the elderly. WMHI and stroke (CVA) volumes were quantified from brain MRI performed on 414 white, male twins born between 1917 and 1927 (average age 72.3 +/- 2.9 years). WMHI, adjusted for age and head size, was significantly correlated with the family history score (r = 0.21, p < 0.001). Dividing the family history scores into quintiles revealed significant differences in WMHI by quintile mean (p < 0.05). Subjects in the highest quintile of family history score had the highest mean WMHI. Recalculation of the family history score, by only counting relatives reported to have had a clinical stroke as a positive event, revealed a nonsignificant correlation with WMHI, but the correlation of the family history score with MRI CVA volume was significant (p < 0.05). Stepwise multivariate analysis including ApoE status, current smoking status, smoking packyear history, Doppler ankle/arm blood pressure ratios, current and long term hypertensive status and current systolic and diastolic pressures indicated that the stroke/hypertension family history score was the single best predictor (p < 0.01) of WMHI volumes. Family history was not an independent predictor of CVA volume. PMID- 10686533 TI - Cost of illness of epilepsy in the US: comparison of patient-based and population based estimates. AB - PURPOSE: To analyze the direct medical costs associated with epilepsy in the US using a cost-of-illness analysis that incorporates both a patient-based approach and a population-based approach. METHODS: Patient-based or 'bottom-up' analysis relied on information provided by a panel of experienced epilepsy clinicians to determine the number and type of medical resources used by individuals with epilepsy. Population-based or 'top-down' analysis relied on cost estimates from a nationally representative sample in the 1987 National Medical Expenditure Survey. RESULTS: The average annual cost per individual in the patient-based analysis was $1,490. The average annual cost per individual in the population-based analysis was $1,510 with average yearly costs per adult of $1,480 and $1,740 per child. New cases of epilepsy are associated with costs of $362 million for the first year of treatment; existing cases of epilepsy are estimated to cost the US nearly $2 billion. CONCLUSIONS: The results of this study indicate incident cases are more than twice as expensive as prevalent cases, and the need to properly care for epilepsy patients is increasingly important in today's health care environment, where the emphasis is on providing effective treatments while simultaneously lowering the costs. PMID- 10686534 TI - Patterns of outcome measurement in Parkinson's disease clinical trials. AB - The study examines the pattern of use and clinimetric properties of clinical endpoints used in randomized trials for Parkinson's disease (PD). Randomized drug trials for PD were identified through a Medline search conducted from January 1966 until August 1998. The endpoints used in these trials were abstracted. Reports examining the clinimetric properties of the disease-specific scales used in these trials were also abstracted. Data regarding the consistency, accuracy, discrimination and feasibility of scales used in at least 10% of trials were determined. One hundred and thirty-seven articles met the inclusion criteria; 70.8% of trials used some clinical scale for PD as an endpoint. The Unified Parkinson's Disease Rating Scale (UPDRS) was the most commonly used scale (32.8%). Factors independently associated with the use of the UPDRS included: the study location in the US, mean age of subjects over 62.7 years and publication after 1994. The UPDRS was more thoroughly studied and superior in most clinimetric domains compared to scales developed earlier. Few studies included generic health status (2.9%) or cognitive measures (16.8%) as secondary endpoints. There have been definite improvements in the area of disease-specific measurement in PD trials since the introduction of the UPDRS. The results of studies that used instruments with poor or unreported clinimetric properties should be critically interpreted. PMID- 10686535 TI - The cladribine trial in secondary progressive multiple sclerosis: A reanalysis. AB - In a recent communication, Goodin [1] analyzes several clinical trials to point out serious flaws in both design and interpretation that may invalidate the conclusions that are drawn. We agree with Goodin [1] that the design of clinical studies, particularly of a disease as complex as multiple sclerosis, is extremely difficult. Indeed, a perfectly designed and executed clinical study is a goal that is never achieved because of the problems inherent in providing care to patients while attempting to evaluate a therapeutic modality. Specifically, in the study of multiple sclerosis, none of the clinical trials of the use of interferons could be considered fully satisfactory: blinding is actually impossible because of the symptoms that are experienced by patients when they receive active drug but not when they receive placebo. It is quite fashionable and not at all difficult to find problems in the conduct of clinical studies; indeed, if all clinical studies with flaws were discarded, there would be no acceptable clinical studies. PMID- 10686536 TI - Evolution of the carcinoembryonic antigen family. structures of CGM9, CGM11 and pregnancy-specific glycoprotein promoters. AB - Earlier studies have demonstrated that the genes of the human carcinoembryonic antigen (CEA) family can be divided into three subgroups, the CEA subgroup (n = 12), the pregnancy-specific glycoprotein (PSG) subgroup (n = 11), and a third subgroup (n = 6). To further characterize the CEA gene family, we have determined the genomic structures of CGM9 and CGM11, analyzed the promoter regions of all eleven PSGs, studied the CGM15-PSG13 intergenic region and the evolutionary relationships beween the CEA family genes. CGM9, a typical CEA subgroup member, was a pseudogene with the exon structure [5'UTR-L-L/N-TM-Cyt-3'UTR]. CGM11 contained a mixture of exons derived from CEA and PSG subgroup genes. The formula of the CGM11 pseudogene was [5'UTR-L-L/N-C-3'UTR]. Thus both genes lacked the IgC2-like domains typically found in CEA subfamily members. The upstream promoter regions of all eleven PSGs were characterized. All PSG promoters lacked the classical TATA and CCAAT elements, but had putative PEA3 box(es), CACCC box(es), a RARE box, and poly (dG-dT) repeats of different lengths. Five PSGs also had an SP1 site. The complete 10-kb intergenic region between CGM15 and PSG13 was sequenced. Clusters of different types of repetitive sequences were seen. The time of divergence of the CEA and PSG subfamilies was estimated to be 107.7 +/- 17.1 million years, or at about the time of human-rodent divergence. Models for the evolution of CEA and PSG and the third family subgroup genes are proposed. PMID- 10686537 TI - Evaluation of serum alkaline DNase activity in treatment monitoring of head and neck cancer patients. AB - Our previously published data on breast cancer suggest that serum alkaline DNase, a known circulating tumour marker, can be used for treatment monitoring of cancer patients. Serum alkaline DNase activities were analyzed in 215 untreated head and neck cancer patients. The enzyme activity ranged from 0.17 to 97.97 IKU/l in untreated cancer patients. Responders (n = 314) showed significantly elevated activity of alkaline DNase as compared to untreated cancer patients (p < 0.001). While non-responders (n = 168) showed comparable activity with untreated cancer patients. Serum alkaline DNase activities were significantly elevated in responders as compared to non-responders (p < 0.001). Our clinical studies during follow-up of patients indicated that the variations in serum alkaline DNase activities in individual patients correlate closely with response to therapy. Serum alkaline DNase also appeared to be useful in predicting treatment response in the long-term follow-up of patients. Serum alkaline DNase was systematically examined as a possible indicator for recurrence in patients under complete remission. In conclusion, serum alkaline DNase may be useful as a treatment monitoring in patients with head and neck malignancies. PMID- 10686538 TI - Endothelial nitric oxide synthase: correlation with histologic grade, lymph node status and estrogen receptor expression in human breast cancer. AB - Nitric oxide is produced by several isoenzymes which are present in many different tissues. We have recently reported the presence of inducible nitric oxide synthase in a breast cancer cell line. The purpose of this study was to examine the distribution of endothelial nitric oxide synthase (eNOS) in a series of human breast tumours. Immunohistochemical investigations demonstrated immunolabelling of tumour cells with the primary antibody, bovine endothelial anti-nitric oxide synthase. Although there was no correlation between eNOS staining and tumour size, there was a significant (p < 0.005) negative correlation (rho = -0.65) between the percentage of tumour cells staining positive for eNOS and the histologic grade of the tumour; there was also a significant (p < 0.05) negative correlation (rho = -0.40) between the percentage of tumour cells staining positive for eNOS and the number of positive lymph nodes. A significant (p < 0.005) positive correlation (rho = 0.63) between the percentage of tumour cells staining positive for eNOS and estrogen receptor (ER) expression by the tumour was also observed. In conclusion, our results demonstrate that eNOS is expressed by human breast tumours and that its presence negatively correlates with histologic grade and lymph node status and positively correlates with ER expression. PMID- 10686539 TI - Serum concentrations of tissue polypeptide antigen in patients with vulvar intraepithelial neoplasia and vulvar cancer. AB - The aim of the present study was to evaluate the clinical usefulness of the cytokeratin tumor marker tissue polypeptide antigen (TPA) in patients with vulvar cancer. This retrospective study comprises 41 patients with vulvar cancer FIGO stages I-III, 17 patients with vulvar intraepithelial neoplasia (VIN) III, and 40 healthy female controls. Serum concentrations of TPA were measured using a microparticle enzyme immunoassay. Results were correlated to clinical data. Median serum concentrations of TPA in healthy female controls, patients with VIN III, and patients with vulvar cancer were 42 U/l (range 12-192), 53 U/l (range 17 127.9) and 57 U/l (range 4.2-423), respectively (Mann-Whitney U test, p = 0.8). Serum concentrations of TPA were not associated with stage of disease, histological grade, and age at the time of diagnosis. In vulvar cancer patients, elevated serum concentrations of TPA prior to therapy were not associated with a shortened disease-free or overall survival (log-rank test: p = 0.5 and p = 0.9, respectively). In a multivariate Cox regression model comprising tumor stage and TPA, tumor stage, but not TPA revealed a statistically significant influence on disease-free (Cox proportional hazard regression model, p = 0.05 and p = 0.6, respectively) and overall (Cox proportional hazard regression model, p = 0.04 and p = 0.8, respectively) survival of patients with vulvar cancer. We conclude that cytokeratin expression, as reflected by serum concentrations of TPA, does not play a role in the natural history of vulvar cancer. The evaluation of serum concentrations of TPA prior to therapy is not recommended. PMID- 10686540 TI - Expression of the tyrosine kinase activity growth factor receptors (EGFR, ERB B2, ERB B3) in colorectal adenocarcinomas and adenomas. AB - The overexpression of three growth factor receptors: epidermal growth factor receptor (EGFR), ERB B2 and ERB B3 was evaluated immunohistochemically in 77 malignant and 15 benign colorectal neoplasms considering clinicopathological variables (histological structure, grade of differentiation, tumor localization, clinical stage of the disease). The relationship between the coexpression of EGFR related proteins in individual patients was also evaluated. EGFR expression was revealed in comparable percentages of colorectal adenoma and in adenocarcinoma cases (80% and 70%) while ERB B2 expression was detectable more frequently in adenoma than in adenocarcinoma cases (87% and 54%). The presence of ERB B3 was observed in a higher percentage of adenocarcinoma than adenoma cases (65% and 40%). There was no correlation between the expression of studied tyrosine kinase receptors and histological grade or Dukes' clinical stage and localization (proximal or distal) of colorectal adenocarcinoma. The incidence of EGFR and ERB B2 expression was higher in tubulovillous (100% for both receptors) than in tubular adenomas (63% and 75%), while the ERB B3 receptor was revealed more frequently in tubular than in tubulovillous neoplasms (50% and 28%). These differences appeared to be statistically nonsignificant. The concomitant expression of two growth factor receptors was observed in a higher percentage of colorectal adenomas than adenocarcinomas, and the coexistence of three growth factors was revealed in comparable percentages in malignant and benign colorectal tumors. Our results support the promotional rather than direct transformational role for the EGFR supergene family in colorectal tumorigenesis. The frequently observed coexpression of more than one EGFR-related protein in colorectal neoplasms indicates the possible cooperation of these receptors in mitogenic signaling transduction, facilitating the development and maintenance of the malignant phenotype. PMID- 10686541 TI - Occurrence of carcinoma-associated antigen 494 in colorectal cancer tissue and in Patients' sera during metastatic disease. AB - CA 494 is a new carbohydrate epitope on a high-molecular-weight mucin-type glycoprotein which has been intensively investigated in pancreatic cancer. In this study, the occurrence of CA 494 was characterized in colorectal cancer tissue and in patients' sera during metastatic disease. CA 494 was detected in cancer tissue from 82% of the 49 patients studied. Serum levels of CA 494 were elevated (>40 U/ml) in 66% of the same patients during metastatic disease (n = 41). The well-established tumor markers carcinoembryonic antigen (CEA) and CA 19 9 were increased (CEA >5 ng/ml; CA 19-9 >37 U/ml) in about 79% of these patients. The correlation of CA 494 with CA 19-9 levels was lower (r = 0.532) than previously reported in pancreatic cancer. PMID- 10686542 TI - Cross-cultural traits for personality of patients with Parkinson's disease in Japan. AB - Recent studies suggest that Parkinson's disease (PD) is associated with particular personality traits. Using Cloningers's Tridimensional Personality Questionnaire (TPQ), Menza and colleagues [1993: Neurology 43:505-508] reported a possible association between PD and a reduced score in the novelty seeking (NS) dimension of the TPQ. We sought to determine whether this association, which was found in a study conducted in the United States, could also be found among Japanese PD patients. We performed personality assessments of 67 Japanese PD patients, using the TPQ test. The results suggest that Japanese PD patients have significantly lower scores in the NS dimension of the TPQ, as well as significantly higher harm avoidance (HA) scores, compared with matched control subjects. Furthermore, the PD patients undergoing treatment for depression using antidepressant drugs scored significantly higher in the HA dimension than PD patients who did not receive antidepressant drug treatment. Our results suggest that the high HA score, and the low NS score in the TPQ test observed in patients with PD, is a cross-cultural phenomenon, although the influence of depression, long-term treatment, and premorbid gene/environmental interactions may also affect these personality traits. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:1 3, 2000. PMID- 10686543 TI - Searching for a locus for schizophrenia within chromosome Xp11. AB - Three gene-rich loci-HS212G6, HSU93305, and HS884M20-within the short arm of the X chromosome have been examined for allelic association with schizophrenia by the transmission disequilibrium test in 70 families of male individuals affected with schizophrenia. Neither the HS212G6 nor HS884M20 was found to be associated with schizophrenia. The HSU93305 locus, however, was significantly associated with schizophrenia (X(2)=17.92, df=3, P<0.001). The HSU93305 locus contains four distinct genes. They code, respectively, for A4 differentiation-dependent protein, triple LIM domain protein, synaptophysin, and calcium channel alpha-1 subunit. It is possible that one of these genes or some loci near to it may predispose a vulnerability to schizophrenia. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:4-7, 2000. PMID- 10686544 TI - Mutation analysis of the inwardly rectifying K(+) channels KCNJ6 (GIRK2) and KCNJ3 (GIRK1) in juvenile myoclonic epilepsy. AB - Genetic factors play a major role in the etiology of idiopathic generalized epilepsy. However, in most syndromes, especially the common ones, multiple genetic factors seem to be involved. Mutations in K(+) channel genes have previously found to be associated with epilepsy both in humans and in mice. The weaver mice phenotype, characterized by ataxia, tremor, male infertility, and tonic-clonic seizures, is caused by a point mutation in the inwardly rectifier K(+) channel gene KCNJ6 (GIRK2). A knockout mouse model deprived of functional KCNJ6 protein is susceptible to spontaneous and provoked seizures without showing the histological signs of neuronal cell death found in the weaver mouse. Thus, the KCNJ6 gene seems to play an important role in seizure control. We therefore performed a mutation analysis of KCNJ6 and the related KCNJ3 gene in 38 patients with juvenile myoclonic epilepsy (JME). Two novel same-sense nucleotide exchanges were identified, but none of these changed the coding sequence. These results do not support a major role for the KCNJ6/KCNJ3 heteromeric receptor in the etiology of JME. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:8-11, 2000 PMID- 10686545 TI - Association analysis between mood disorder and monoamine oxidase gene. AB - To ascertain whether mood disorders, including bipolar and unipolar, are genetically associated with the monoamine oxidase A (MAOA) or monoamine oxidase B (MAOB) gene in the Chinese population, 132 cases of mood disorder and 88 normal controls were genotyped for the MAOA(CA)n, MAOB(GT)n, and MAOB(TG)n loci by the method of amplification fragment length polymorphism. Among 132 cases with mood disorder, eight alleles (size: 112-126 bp) of locus MAOA(CA)n, 12 alleles (size: 168-198 bp) of locus MAOB(GT)n, and nine alleles (size: 195-213 bp) of locus MAOB(TG)n were observed. Comparison of the allele frequency of the three loci showed no difference between mood disorder cases and normal controls on average. When each group was stratified into several subgroups, significant differences were found. On the MAOA(CA)n locus, the frequency of 116 bp allele was higher in the female bipolar disorder cases (0.2581) compared with that in the female unipolar disorder patients (0.1154) (Z=2.15, p<0. 05). On the MAOB(GT)n locus, the frequency of 180 bp allele was higher in bipolar disorder patients (0.1579) than that in normal controls (0.0678) (Z=2.05, p<0.05). The frequency of this allele was even higher in female bipolar disorder patients (0.1719) than that in female normal controls (0.0541). On the MAOB(TG)n locus, the frequency of 205 bp allele was higher in female bipolar disorder patients (0.6406) than that in female normal controls (0.4375) (Z=2. 17, p<0.05). For the unipolar disorder patients, the frequency of this allele was higher in female cases (0.5222) than that in male cases (0.1818) (Z=3.49, p<0.05). As for association studies, significant association between bipolar disorder and MAOB gene was detected. For the 180 bp allele of MAOB(GT)n, the relative risk (RR) of biploar versus normal control was 2.58 (p<0.05), and the RR of female bipolar disorder versus female normal control was 3.63 (p<0. 05). For the 205 bp allele of MAOB(TG)n, the RR of female bipolar disorder versus female normal control was 2.29 (p<0.05). Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:12-14, 2000. PMID- 10686546 TI - Chromosome 22q11 deletions are not found in autistic patients identified using strict diagnostic criteria. IMGSAC. International Molecular Genetics Study of Autism Consortium. AB - A group of 103 subjects with a strict diagnosis of autism were tested for deletion of band q11.2 on the long arm of chromosome 22. No deletions were found, indicating that when a patient has been diagnosed with autism using strict and consistent criteria, in the absence of other indications, it is unlikely that this individual will have a 22q11 deletion. Testing for 22q11 deletions is therefore unlikely to be necessary in these patients. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:15-17, 2000. PMID- 10686547 TI - Suggestive evidence of a locus on chromosome 10p using the NIMH genetics initiative bipolar affective disorder pedigrees. AB - As part of a four-center NIMH Genetics Initiative on Bipolar Disorder, a genome screen using 365 markers was performed on 540 DNAs from 97 families, enriched for affected relative pairs. This is the largest uniformly ascertained and assessed linkage sample for this disease, and includes 232 subjects diagnosed with bipolar I (BPI), 32 with schizo-affective, bipolar type (SABP), 72 with bipolar II (BPII), and 88 with unipolar recurrent depression (UPR). A hierarchical set of definitions of affected status was examined. Under Model I, affected individuals were those with a diagnosis of BPI or SABP, Model II included as affected those fitting Model I plus BPII, and Model III included those fitting Model II plus UPR. This data set was previously analyzed using primarily affected sib pair methods. We report the results of nonparametric linkage analyses of the extended pedigree structure using the program Genehunter Plus. The strongest finding was a lod score of 2.5 obtained on chromosome 10 near the marker D10S1423 with diagnosis as defined under Model II. This region has been previously implicated in genome-wide studies of schizophrenia and bipolar disorder. Other chromosomal regions with lod scores over 1.50 for at least one Model Included chromosomes 8 (Model III), 16 (Model III), and 20 (Model I). Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:18-23, 2000 PMID- 10686548 TI - Potential panic disorder syndrome: clinical and genetic linkage evidence. AB - This paper reports evidence for a possible "chromosome 13 syndrome," which includes panic disorder, kidney or bladder problems, serious headaches, thyroid problems (usually hypothyroid), and/or mitral valve prolapse (MVP). In the course of a genetic linkage study of panic disorder, we noted these medical conditions in individual family members. (We were blind to family relationships and marker data.) We hypothesized that there may exist a subgroup of panic families with these medical conditions, which for simplicity we called it the "syndrome." Subsequently we reclassified the families as with or without the "syndrome" and extended the phenotype for analysis to include the above medical conditions. All these classifications were also done before the analysis and blind to marker data. We then examined our linkage results, looking for significant differences between families with and without the "syndrome" (using several definitions of the "syndrome")-i.e., testing for genetic heterogeneity. When the families with and without bladder/kidney problems were separated from each other, one marker D13S779 (ATA26D07)-yielded a lod score of over 3 in the families with bladder/kidney problems. This lod score went up to 4.2 in these families when we diagnosed any individual with any one of the "syndrome" conditions as affected. These results were statistically significant even after applying an extremely overconservative Bonferroni correction for multiple tests. We present these results in order to alert other investigators working on panic disorder, for replication. If replicated, one may hypothesize that a candidate gene for the syndrome should be expressed in CNS, kidney, gut, thyroid, etc. We also noted that two independent studies report recent linkage findings between schizophrenia and the same region on chromosome 13. No connection between schizophrenia and panic disorder has ever been reported. Finally, we suggest that genetic studies of psychiatric disorders might prove more fruitful if phenotypes were expanded to include possible manifestations of the disorder in medical (non-mental) symptoms. Am. J. Med. Genet.(Neuropsychiatr. Genet.) 96:24-35, 2000. PMID- 10686549 TI - Genetic associations with clinical characteristics in bipolar affective disorder and recurrent unipolar depressive disorder. AB - Genetic factors may be associated with disease subtype as well as susceptibility. We have therefore typed polymorphisms at the serotonin transporter, dopamine receptor, tryptophan hydroxylase, tyrosine hydoxylase, and monoamine oxidase A (MAOA) loci in 139 unipolar and 131 bipolar patients and investigated associations with gender, number of episodes, age of onset, history of psychotic symptoms, history of suicidal behavior, and history of substance abuse. In bipolar subjects, the promoter variable number tandem repeat (VNTR) allele 132 of MAOA was associated with history of suicide attempts, P = 0.029, particularly in females, P = 0.006. The Fnu4HI allele 1 of MAOA was also associated with history of suicide attempts in females, P = 0.0162. The serotonin transporter promoter allele 2 was associated with increasing number of manic episodes, P = 0.02, and history of psychotic symptoms, P = 0.0243. One significant association was found in the unipolar group: dopamine D2 receptor promoter allele 2 with history of psychotic symptoms, P = 0. 0165. We have tested multiple loci for a variety of different clinical variables and performed 228 tests of significance in total. It is possible that these preliminary findings are type 1 errors, because one would expect 11 of the 228 tests to reach a nominal significance level of P < 0.05 by chance alone if all the tests were independent. The associations with the MAOA and serotonin transporter loci are consistent with previous data suggesting associations with susceptibility to bipolar affective disorder. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:36-42, 2000 PMID- 10686550 TI - Analysis of linkage disequilibrium in gamma-aminobutyric acid receptor subunit genes in autistic disorder. AB - Autistic disorder (AD) is a neurodevelopmental disorder characterized by abnormalities in behavior, communication, and social interactions and functioning. Recently, Cook et al. reported significant linkage disequilibrium with an AD susceptibility locus and a marker, GABRB3 155CA-2, in the gamma aminobutyric acid(A) (GABA(A)) receptor beta3-subunit gene on chromosome 15q11 q13. This linkage disequilibrium was detected using a multiallelic version of the transmission/disequilibrium test (TDT) in a sample of nuclear families having at least one child with autistic disorder. In an attempt to replicate this finding we tested for linkage disequilibrium with this marker, as well as with three additional markers in and around the GABA(A) receptor beta3-subunit gene, in an independent, clinically comparable set of AD families. Unlike Cook et al., we failed to detect significant linkage disequilibrium between GABRB3 155CA-2 and AD in our sample. We did, however, find suggestive evidence for linkage disequilibrium with a marker, GABRB3, approximately 60 kb beyond the 3' end of beta3-subunit gene. This finding lends support for previous reports implicating the involvement of genes in this region with AD. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:43-48, 2000 PMID- 10686551 TI - No evidence for a major susceptibility locus for juvenile myoclonic epilepsy on chromosome 15q. AB - Juvenile myoclonic epilepsy (JME) is a distinct epileptic syndrome with a complex mode of inheritance. Several studies found evidence for a locus involved in JME on chromosome 6 near the HLA region. Recently, Elmslie et al. [1997] reported evidence of linkage in JME to chromosome 15q14 assuming a recessive mode of inheritance with 50% penetrance and 65% linked families. The area on chromosome 15q14 encompasses the location of the gene for the alpha-7 subunit of the nicotinic acetylcholine receptor. This could fit the hypothesis that there are two interacting loci, one on chromosome 6 and on chromosome 15 or that there is genetic heterogeneity in JME. In an independent dataset of JME families, we tested for linkage to chromosome 15 but found little evidence for linkage. Moreover, families with more than one family member affected with JME provide a lodscore of 3.4 for the HLA-DR/DQ haplotype on chromosome 6. The lodscore for these same families on chromosome 15q14 is <-2 assuming homogeneity and the maximum lodscore is 0.2 assuming alpha =.25. Only one of these families has a negative lodscore on chromosome 6 and a positive lodscore of 0.5 on chromosome 15q14. Our results indicate that this possible gene on chromosome 15 plays at most a minor role in our JME families. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:49-52, 2000. PMID- 10686552 TI - Analysis of the serotonin transporter gene linked polymorphism (5-HTTLPR) in anorexia nervosa. AB - Previous studies have demonstrated aberrant expression of serotonin in individuals with an eating disorder. Given this the serotonin transporter gene (5 HTT) is a strong candidate to contribute to the genetic component of the aetiology of eating disorders. To determine the role of this particular gene in the susceptibility to anorexia nervosa (AN) we have examined a tandemly repeated sequence close to the promotor region of the 5-HTT gene, which is represented by a long (L) and short (S) variant. Previous studies have shown that the transcriptional activity of the 5-HTT gene differs significantly between these two alleles. A group of 138 Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria AN patients and 90 controls were genotyped at the 5-HTT gene linked polymorphism (5-HTTLPR). Statistical analysis showed no significant difference in allele or genotype frequencies between the two groups. These data suggest that there is no association between 5-HTTLPR genotype and susceptibility to AN, in our population. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:53-55, 2000. PMID- 10686553 TI - Association of polymorphism of serotonin 2A receptor gene with suicidal ideation in major depressive disorder. AB - There is evidence indicating that density of 5-HT2A receptors is altered in brain regions of depressed suicide victims and in platelets of suicidal subjects with major depression or schizophrenia. Recent studies have also shown an association between the allele C of 102T/C polymorphism in the 5-HT2A receptor gene and schizophrenia. The present investigation tested the hypothesis that the observed changes in 5-HT2A receptor density in platelets of patients with major depression are a trait rather than state phenomenon and are associated with the 102 C allele in 5-HT2A receptor gene in a sample of 120 patients with major depression and a group of 131 control subjects comparable with respect to age, sex, and ethnic background. The allele and genotype frequencies of 102T/C polymorphism in 5-HT2A receptor gene were compared between patients and control subjects and between suicidal and non-suicidal patient groups. The major finding of this study was a significant association between the 102 C allele in 5-HT2A receptor gene and major depression, chi(2) = 4.5, df = 1, P = 0.03, particularly in patients with suicidal ideation, chi(2) = 8.5, df = 1, P < 0.005. Furthermore, we found that patients with a 102 C/C genotype had a significantly higher mean HAMD item 3 score (indication of suicidal ideation) than T/C or T/T genotype patients. Our results suggest that the 102T/C polymorphism in 5-HT2A receptor gene is primarily associated with suicidal ideation in patients with major depression. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:56-60, 2000. PMID- 10686554 TI - Anticipation in schizophrenia and bipolar disorder controlling for an information bias. AB - Anticipation was investigated in schizophrenia (SZ) and bipolar disorder (BP) while addressing several biases in 18 large families (154 subjects) from Eastern Quebec densely affected by SZ, BP, or both over three generations. In particular, we controlled for an information bias using a measure of quality and quantity of clinical information (QOI) concerning the subjects' illness. Otherwise, spurious anticipation could have arisen because we found that QOI varied with the generations as well as with the severity of illness. Although anticipation was investigated separately for SZ and BP, both disorders were also included in one analysis that tested anticipation under the unitary hypothesis that the SZ and the BP spectrums represent a continuum of severity of the same disease. Age of onset (AOO) and five indices of severity were tested for anticipation. Two statistics were used: the difference in the mean AOO or severity between two successive generations, and the mean difference in parent-offspring pairs (POP). The study led to four main findings: 1) the choice of the statistics greatly influenced the results, POP yielding systematically greater biased estimates; 2) for SZ and BP, the evidence for anticipation with the five severity indices vanished after controlling for QOI; 3) as regards AOO a decrease of 8.6 years, p = 0.0001, and 5.3 years, p = 0.009 in AOO was found for SZ between Generations 1 2, and 2-3, respectively, despite controlling for QOI and addressing all biases; and 4) conversely for BP, anticipation with AOO may be due to censoring. Findings suggest that future anticipation studies should also control for QOI. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:61-68, 2000. PMID- 10686555 TI - Peek-a-boo fragile site at 16d associated with Tourette syndrome, bipolar disorder, autistic disorder, and mental retardation. AB - Five patients with a fragile site at 16q22-23 and neuropsychiatric disorders are reported. Three of five had Tourette disorder, three had mental retardation, two had bipolar disorder, and one had autistic disorder. During our attempts to study the fragile sites in more detail we were unable to reproduce the fragile sites found several years earlier. The potential relationship between the fragile sites and the neuropsychiatric disorders in these patients is discussed. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:69-73, 2000. PMID- 10686556 TI - Evaluation of linkage of markers on chromosome 6p with schizophrenia in Taiwanese families. AB - Previous studies have indicated possible linkage of schizophrenia with chromosome 6p21-24. In an attempt to replicate these findings, we studied the linkage of schizophrenia with nine markers on chromosome 6p21-24 in 39 Taiwanese schizophrenic nuclear families with at least two affected siblings. Two diagnostic models (narrow: Diagnostic and Statistical Manual of Mental Disorders IV schizophrenia only; and broad: including schizophrenia, schizoaffective, and other nonaffective psychotic disorders) were used to define the disease phenotypes. With the broad and narrow diagnostic models, the marker D6S296 produced maximum two-point lod scores of 1.46 (straight theta = 0.2) and 1.35 (straight theta = 0. 2), respectively, in the recessive inheritance model. Assuming locus heterogeneity, a multipoint lod score of 0.85 was obtained between markers D6S296 and D6S277 under the narrow/recessive model. Maximum nonparametric lod scores of 1.25 ( p= 0.09) and 1.36 (p = 0.08) were observed, but still not statistically significant, at D6S296 in the narrow and broad diagnostic models, respectively. Both two-point analysis of the dominant model (lod score 0.85) and nonparametric analysis (lod score 1.25) showed a mild peak lod score appeared at marker D6S 285 as well. The results add some support to the suggestive linkage of schizophrenia with markers in the regions of chromosome 6p22 and 6p24 in an ethnically distinct Taiwanese sample. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:74-78, 2000. PMID- 10686557 TI - Lack of association between temporal lobe epilepsy and a novel polymorphism in the alpha 2 subunit gene (ATP1A2) of the sodium potassium transporting ATPase. AB - Genetic linkage studies in rodents and humans have identified specific chromosomal regions harboring seizure susceptibility genes. We have identified a novel polymorphism in the human alpha 2 subunit gene (ATP1A2) of the sodium potassium transporting ATPase (NaK-pump), a candidate gene for human temporal lobe epilepsy (TLE) based on its chromosomal location and function in ion homeostasis. The polymorphism consists of a four base pair insertion 12 base pairs upstream of the start of exon 2. We performed an association study between this polymorphism and TLE. Our study did not find a significant difference in the frequency of this polymorphism between TLE patients and controls, indicating that this variation is not a major susceptibility factor. However, since the number of patients studied so far is small and the functional consequence of the polymorphism is unknown, the variation may yet be found to play a minor role in increased risk for seizure susceptibility. In contrast to the findings in TLE patients and controls, we did find a significant difference in the frequency of the variation between African Americans and persons of European descent. This finding demonstrates the potential effect of population stratification on studies of this type and supports the growing use of parental and familial samples for controls in association studies. Further study of this polymorphism is warranted as it may be involved in other disease processes for which there are known ethnic specific susceptibilities. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:79-83, 2000. PMID- 10686558 TI - Serotonin-2A receptor gene is not associated with symptomatology of schizophrenia. AB - The serotonin receptor type 2A (5-HT2A) is a primary candidate for involvement in major psychoses. Polymorphisms within the 5-HT2A gene have recently been reported to be associated with a variety of psychopathological conditions. In the present study, we investigated the potential influence of the T102C polymorphism on the psychopathology of schizophrenia. One hundred eighty-eight inpatients affected by schizophrenia (DSM-III-R) were assessed by the Operational Criteria checklist for psychotic illness (OPCRIT) and were typed for their 5-HT2A variants by PCR techniques. Mania, depression, delusion and disorganization were the four symptomatologic factors previously derived from our psychotic population that were used to define phenotype in our sample. Genetic variants of the polymorphism under study were not associated with these symptomatologic factors, and consideration of possible stratification effects such as sex and age of onset did not reveal any association either. Our results do not, therefore, support the hypothesis that the serotonin receptor 2A gene is a liability factor for the symptomatology of schizophrenia as defined by the OPCRIT checklist. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:84-87, 2000. PMID- 10686559 TI - Analysis of the polymorphic (GT)(n) repeat at the dopamine beta-hydroxylase gene in Spanish patients affected by schizophrenia. AB - The presence of a polymorphic (GT)(n) repeat, a microsatellite repeat, at the human dopamine beta-hydroxylase (DBH) gene had been previously investigated in healthy people and in schizophrenic patients. The different DBH genotypes had been found to be associated to different DBH biochemical function, but no differences were found in the allelic and genotype frequencies between schizophrenic and control groups. To further clarify the potential involvement of the variation at the DBH gene in schizophrenia we have studied the DBH (GT)(n) repeat in a sample of 47 Spanish schizophrenic patients, in their healthy relatives (n = 72), and in a control population (n = 74). We have been able to identify five different variants of the DBH gene (A1, A2, A3, A4, A5) in the different groups. Subsequent statistical analysis revealed that the genotypes as well as the allele frequencies did not differ significantly among schizophrenic patients and the control population. Interestingly, the allelic variant A2 and the genotype A4/A2 were significantly more frequent in schizophrenic patients as compared with their healthy relatives. However, the association of the A2 allele with schizophrenia was not supported by the haplotype relative risk analysis of transmitted versus nontransmitted alleles. Therefore, although it will be important to extend the present analysis in a larger sample of schizophrenic patients and controls, our results suggest that the (GT)(n) does not seem to play a major role in the genetics of schizophrenia at least in this group of Spanish schizophrenic patients. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:88-92, 2000. PMID- 10686560 TI - Selecting early onset MDD probands for genetic studies: results from a longitudinal high-risk study. AB - Recent studies have found high rates of familial aggregation of major depression (MDD) in relatives of depressed children coming for treatment, leading investigators to suggest that probands for genetic studies of MDD should be selected from samples of depressed children being brought for treatment. Implicit in this recommendation is the assumption that childhood and adult depression are similar disorders. This assumption in turn implies that children with prepubertal or adolescent onset depression are at high risk for having recurrent episodes of MDD that continue into adulthood. The data supporting this latter hypothesis, however, is limited and contradictory. In this article we report results from a high-risk longitudinal family study in which we explored the recurrence and continuity into adulthood of prepubertal or adolescent onset MDD in offspring who were at high or low risk for MDD, by virtue of their parental depression status. One hundred eighteen offspring from 55 families in which one or more parents had MDD and 50 offspring from 21 families in which neither parent had MDD were followed for more than 10 years (all offspring were 20 years or older at the end of follow-up time) and blindly reassessed using a semistructured diagnostic instrument. Offspring with childhood/adolescent onset MDD were at significantly greater risk for recurrence in adulthood (after age 25) as compared with offspring without an onset of childhood/adolescent MDD, if they had a history of parental MDD. In contrast, among offspring without a history of parental MDD, those with childhood/adolescent onset MDD were at no greater risk for continuing to have MDD in adulthood (after age 25) than those without childhood/adolescent onset MDD. Moreover, there was a trend for offspring with childhood/adolescent onset MDD to be at greater risk for recurrence after age 25 if they had a history of parental MDD, as compared with offspring without a history of parental MDD (60 vs. 18%). We conclude that childhood/adolescent onset MDD is a heterogeneous disorder, with family history of MDD appearing to define a subtype of childhood/adolescent onset MDD that is recurrent and continues into adulthood. Our findings suggest that caution should be exercised in selecting depressed children and adolescents brought for treatment as probands in genetic studies of early onset MDD. A conservative strategy would be to select only those depressed children and adolescents with a family history of MDD and reassess the treated sample as they mature, ensuring that they go on to have MDD in adulthood. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:93-101, 2000 PMID- 10686561 TI - COMT and DRD3 polymorphisms, environmental exposures, and personality traits related to common mental disorders. AB - In a community sample of 2,327 Caucasians, we tested the hypotheses that polymorphisms in the COMT and DRD3 genes are associated with personality traits conferring vulnerability to anxiety, depression, or alcohol misuse, or with current symptoms of these; and that the association is stronger in persons who also have been exposed to stressor experiences. To conserve resources and to allow replication, the genetic analysis was undertaken in two stages. For the COMT polymorphism, no statistically significant associations were found in the first sample of 862 persons. The remainder of the sample was therefore not analysed for that gene. For the DRD3 polymorphism, those in the first sample with at least one of the Ser(9) alleles had significantly higher scores in neuroticism (p=0.006) and behavioral inhibition (p=0.003). There was a trend, failing to meet the 1% significance criterion, for those with this genotype also to have higher depression and anxiety. The groups did not differ in alcohol use. In persons with the Ser(9) allele who were also exposed to stressors, there was a higher level of depression at the 5% level; and the depression level was higher in homozygotes. But when the remainder of the sample (1,465) was analysed, none of the associations reached statistical significance. We conclude that neither the COMT nor DRD3 polymorphisms are associated with anxiety, depression, or alcohol abuse. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:102-107, 2000 PMID- 10686562 TI - Integration of genetic maps by polynomial transformations. AB - Currently available genetic maps differ in a variety of basic features; in particular, with respect to the total length of the genome. Consequently, the question arises as to the extent to which genetic maps are compatible to each other, as well as to the methods with which genetic maps can be transformed into one another. We propose a set of nonlinear, polynomial transformations that enable the integration of genetic maps at a sufficiently high overall precision. Our analysis of six major, publicly available maps, and iteratively optimized polynomials of up to degree 5, yielded differences of 90% of points. Similarly, we determined, at a slightly worse overall fit, those polynomials that enabled the reconstruction of sex-specific recombination estimates from sex-averaged data. Our results suggest that polynominal transformations may become a valuable extension of standard map construction methods due to a rapid integration of newly developed markers into existing maps. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:108-113, 2000. PMID- 10686563 TI - Linkage study of two polymorphisms at the dopamine D3 receptor gene and attention deficit hyperactivity disorder. AB - Data from animal studies suggest that the dopamine D3 receptor gene may have a role in locomotion and behavioral regulation. Therefore, this gene has been suggested as a candidate for attention-deficit hyperactivity disorder (ADHD). The dopamine D3 receptor gene (DRD3) has two common polymorphisms, one in exon I that changes a Serine to Glycine (Ser9Gly) and alters the recognition site for the restriction enzyme MscI [Lannfelt et al., 1992]. The other common polymorphism is located in intron 5 and results in the change of a restriction site for MspI [Griffon et al., 1996]. We investigated the possibility of linkage of the dopamine D3 receptor gene in 100 small, nuclear families consisting of a proband with ADHD, their parents, and affected siblings. We examined the transmission of the alleles of each of these polymorphisms and the haplotypes of both polymorphisms using the transmission disequilibrium test [Spielman et al., 1993]. We did not observe biased transmission of the alleles at either polymorphism or any haplotype. Our findings using this particular sample do not support the role of the dopamine D3 gene in ADHD. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:114-117, 2000. PMID- 10686564 TI - Polymorphisms of the sigma(1) receptor gene in schizophrenia: An association study. AB - Possible involvement of sigma receptors in the pathogenesis of schizophrenia has been suggested. In this study we searched systematically for polymorphisms in the 5'-franking region of the sigma(1) receptor. Genetic variation in this region could reduce the expression of the gene, and this suggestion is compatible with findings of reduced sigma binding sites in several cortical regions of schizophrenia. We confirmed G-241T and G-240T polymorphisms; these two consecutive polymorphisms were resolved to be in complete linkage disequilibrium with each other by single-strand conformation polymorphism (SSCP) analysis. We also identified the A61C (Gln2Pro) polymorphism, which was in almost complete linkage disequilibrium with G-241T/G-240T. There was no significant difference in the distribution of alleles or overall genotypes of the polymorphisms between schizophrenic patients (n = 129) and controls (n = 140). We found slight increased homozygosity for T-241/T-240 and C61 in patients compared with controls using multiple comparison (p = 0. 045). However, the significance did not remain when a Bonferroni correction was made (p = 0.135). These results do not support that the sigma(1) receptor gene plays a major role in the pathogenesis of schizophrenia. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:118-122, 2000. PMID- 10686565 TI - Lack of association between serotonin transporter gene promoter variants and autistic disorder in two ethnically distinct samples. AB - Family-based studies performed to date provide conflicting evidence of linkage/association between autistic disorder and either the "short" [Cook et al., 1997: Mol Psychiatry 2:247-250] or the "long" [Klauck et al., 1997: Hum Mol Genet 6:2233-2238] allele of a polymorphic repeat located in the serotonin transporter (5-HTT) gene promoter region, affecting 5-HTT gene expression [Lesch et al., 1996: Science 274:1527-1531]. The present study was designed to assess linkage and linkage disequilibrium in two new ethnically distinct samples of families with primary autistic probands. The 5-HTT promoter repeat was genotyped in 54 singleton families collected in Italy and in 32 singleton and 5 multiplex families collected in the U.S.A., yielding a total sample of 98 trios. Linkage/association between 5-HTT gene promoter alleles and autistic disorder was assessed using the transmission/disequilibrium test (TDT) and the haplotype-based haplotype relative risk (HHRR). Both the Italian and the American samples, either singly or combined, displayed no evidence of linkage/association between 5-HTT gene promoter alleles and autistic disorder. Our findings do not support prominent contributions of 5-HTT gene variants to the pathogenesis of idiopathic infantile autism. Heterogeneity in pathogenetic mechanisms underlying the disease may require that linkage/association studies be targeted toward patient subgroups isolated on the basis of specific biochemical markers, such as serotonin (5-HT) blood levels. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:123-127, 2000. PMID- 10686566 TI - Autistic symptoms among children and young adults with isodicentric chromosome 15. PMID- 10686567 TI - Shining a light on the black box of the review process at Research in Nursing & Health. PMID- 10686568 TI - Families and hospitalized elders: A typology of family care actions. AB - The extensive care provided by families to their elderly relatives in the home is well documented. Although family caregiving is likely to be continued during hospitalization of elderly relatives, limited research has been conducted to address the nature of family care for hospitalized elders. The main purpose of this qualitative study was to refine the content domain of family care for hospitalized elders. Altogether 25 interviews were done. Of the 16 participants, 6 were family members, 6 were patients, and 4 were nurses; 7 participants were interviewed once and 9 participants were interviewed twice. Qualitative analysis based on Lofland and Lofland's (1984, 1995) approach resulted in the identification of three major content domains: family members providing care to the patient, working together with the health care team, and taking care of themselves. This typology suggests a shift of research in this area from its current focus on family needs to a view of family caregivers as partners with the health care team. PMID- 10686569 TI - Positive and negative outcomes of anger in early adolescents. AB - The purposes of this study were to examine symptom patterns and diminished general well-being as negative outcomes and vigor and change as positive outcomes of trait and state anger via two structural equation models. In a school auditorium, a convenience sample of 141 boys and girls, ages 12-14 years, responded to the Trait Anger Scale and the State Anger Scale and to instruments measuring general well-being, symptom patterns, vigor, and change. In the negative outcome model, results indicated that diminished general well-being and increased symptom patterns were outcomes of trait anger and state anger in early adolescents. In the positive outcome model, contrary to expectation, less vigor and less inclination to change were outcomes of trait anger in early adolescents, while state anger had no appreciable influence on the same variables. The findings suggest that anger, particularly trait anger, had a negative influence on the outcome variables studied. PMID- 10686570 TI - Sense of coherence and illness appraisal in older women's quality of life. AB - The purpose of this descriptive, correlational study was to test a conceptual model of proposed relationships between physical health limitation, the sense of coherence, illness appraisal, and quality of life in a sample of 137 older women. The typical respondent was 76, widowed, with an income less than $12,000, and several health problems. Hierarchical multiple regression analysis indicated that physical health limitation, particularly symptom bother and functional health, had a significant negative influence on quality of life. However this effect was mediated by sense of coherence and illness appraisal. Regardless of the level of symptoms or functional health, women with higher sense of coherence and more positive illness appraisals had higher levels of quality of life. The findings support the proposed model and further our understandings regarding the protective role of personality resources in perceived quality of life in older women with chronic illnesses. PMID- 10686571 TI - Continuous handrail support, oxygen uptake, and heart rate in women during submaximal step treadmill exercise. AB - Past research suggests that continuous handrail support during exercise attenuates physiologic responses to exercise and reduces aerobic benefits; however, this phenomenon has not been systematically studied in women exercising on the step treadmill. The effects of three levels of handrail support (continuous light, continuous very light, or no handrail support) on oxygen uptake and heart rate during step treadmill exercise were examined in 15 healthy women. Measures were obtained during 6 bouts of exercise, 3 bouts at 25 steps/min followed by 3 bouts at 33 steps/min. At both step rates, mean oxygen uptake was significantly reduced during continuous light and continuous very light handrail support as compared with no handrail support, and mean heart rate was significantly reduced during continuous light versus no handrail support. At 25 steps/min only, mean heart rate was significantly reduced during continuous very light versus no handrail support. Findings indicate that women who use even continuous light or continuous very light handrail support attenuate physiologic responses during step treadmill exercise, thereby reducing aerobic requirements and gaining suboptimal benefits from exercise. PMID- 10686572 TI - The reliability and validity of two health status measures for evaluating outcomes of home care nursing. AB - The reliability, validity, and sensitivity of the Medical Outcome Study Short Form (SF-36) and the Quality of Life Profile: Senior Version (QOLPSV) for measuring outcomes of home care nursing were evaluated. Data were collected from 50 clients receiving home care nursing services. Twenty-two registered nurses and six registered practical nurses collected client and nursing data on each home visit. Client baseline and outcome measures were collected by two independent evaluators at admission and discharge from the home care service. Internal consistency reliability ranged from.76 to.94 for the eight subscales of the SF 36. Internal consistency reliability ranged from.47 to.82 for the nine subscales of the QOLPSV. The subscales of both instruments had minimal problems with missing responses. The SF-36 was found to be more sensitive than the QOLPSV to change over time. In addition, the subscales of the SF-36 were found to be more sensitive than the subscales of the QOLPSV to several of the nursing variables, such as intensity of the client's nursing condition and skill mix. PMID- 10686573 TI - Test of a model of psychosocial resources, stress, and health among undereducated adults. AB - The purpose of this study was to test a model drawn from the modeling and role modeling theory depicting relationships among psychosocial resources, perceived stress, and health for undereducated adults. A purposive sample of 171 adults enrolled at an urban adult education center completed several self-report measures: Modified Erikson Psychological Stage Inventory, Basic Need Satisfaction Inventory, Perceived Stress, and Positive Health Index. Based on a structural equation modeling analysis, psychosocial development and basic need satisfaction had significant direct effects on health, with the expected positive signs. Psychosocial development had the strongest direct effect on health and also had a strong direct effect on basic need satisfaction and an indirect effect on health. Support for the hypothesized model has important implications for nursing and other community-based care provider interventions regarding health, including strengthening psychosocial resources. PMID- 10686574 TI - Personal and social determinants of rural nurses' willingness to care for persons with AIDS. AB - The purpose of this study was to investigate the individual and social determinants of rural nurses' willingness to care for people with AIDS (PWAs). Willingness to care was viewed as a function of nurses' personal attitudes about AIDS care and PWAs; the influence of normative (significant others), comparative (the nursing profession), and generalized (the rural community) reference group norms on these attitudes; and how much importance respondents placed on membership in these reference groups. Responses to a mailed questionnaire from 615 rural nurses were analyzed. Individual determinants were nurses' feelings of preparedness and favorable attitudes about their personal safety when administering care. Social determinants were the degree of upset of respondents' significant others about their caring for AIDS patients and favorable attitudes of the respondents about professional and social concerns related to AIDS. PMID- 10686575 TI - Threats to validity in randomized clinical trials. AB - The purposes of this article are to present an overview of randomized clinical trials (RCTs) and describe some of the methodological problems inherent in using RCTs in nursing research. Many nursing intervention studies are fraught with problems that defy the stringent control criteria required for RCTs, leading to biased estimates of intervention efficacy. Five threats to validity in RCTs are presented, including problems related to (a) differential dropout, (b) random assignment, (c) identifying and maintaining an adequate control condition, (d) nonadherence to research protocols, and (e) assessment of clinically meaningful change. Three strategies are recommended for addressing some of the problems posed by RCTs and improving inference. PMID- 10686578 TI - Effects of pro-inflammatory cytokines on apolipoprotein E secretion by a human astrocytoma cell line (CCF-STTG1). AB - Apolipoprotein (apo) E has been implicated in Alzheimer's disease; however, little is known about the regulation of its secretion in astrocytes. To investigate the effects of pro-inflammatory cytokines such as interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN gamma) on apoE secretion by CCF-STTG1 cells, a sensitive and specific double sandwich Enzyme-Linked ImmunoSorbent Assay (ELISA) was developed. Using a monoclonal anti-human apoE antibody as the capture antibody, this assay was carried out with commercially available reagents. The assay had a sensitivity of 0.013 ng per well, within-run and between-run variation coefficients of 6.0 and 8.6 per cent respectively. There was no cross-reactions between antibodies used and apoAI, apoAII, apoB, apoCI, apoCII and apoCIII. Low apoE concentrations were assessed using a serum-free HepG2 culture medium as secondary calibrator, containing 59 microg l(-1) of apoE. In serum-free medium, CCF-STTG1 cells secreted apoE, the accumulation of which in the cell medium increased linearly with time (27 microg per 48 h). After 48 h of incubation, apoE secretion was inhibited by TNF-alpha but not affected by IL-1beta and IFN-gamma. However, the effect of regulatory factors may depend upon culture conditions since in the presence of 10 per cent fetal calf serum, IFN-gamma significantly inhibited apoE secretion. Thus, apoE secretion by CCF-STTG1 cells is inhibited by specific pro inflammatory cytokines. This new apoE ELISA presents the great advantage of using commercially available reagents which permit inter-laboratory comparability of results, involves relatively low cost and is adaptable for the measurement of low levels of apoE. PMID- 10686577 TI - Degradation of focal adhesion proteins during nocodazole-induced apoptosis in rat 1 cells. AB - Nocodazole, a microtubule-disrupting agent, induced apoptosis in Rat-1 cells, as indicated by changes in cell morphology, DNA fragmentation, and eventual cell death. During nocodazole-induced apoptosis, normally flat cells became rounded in shape and detached from the extracellular matrix. These morphological changes appeared to be closely associated with degradation of focal adhesion proteins, including p130cas, p125(FAK) and paxillin. p130cas was also degraded in cells treated with staurosporine or etoposide, suggesting that degradation of focal adhesion proteins is a characteristic feature of apoptosis. Nocodazole-induced apoptosis was antagonized by Bcl-2: degradation of focal adhesion proteins was suppressed and cell viability was enhanced in bcl-2 over-expressing cells, even in the presence of nocodazole. Further study of the molecular mechanism of Bcl-2 activation should provide an understanding of the apoptosis induced by disruption of the microtubule network. PMID- 10686579 TI - Modulatory effects of garlic and neem leaf extracts on N-methyl-N'-nitro-N nitrosoguanidine (MNNG)-induced oxidative stress in Wistar rats. AB - The effects of garlic and neem leaf extracts on lipid peroxidation and antioxidant status during administration of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a carcinogenic nitrosamine were evaluated in male Wistar rats. Extracts of garlic and neem leaf were administered orally for five consecutive days before intraperitoneal injection of MNNG. Enhanced lipid peroxidation in the stomach, liver and circulation of MNNG-treated rats was accompanied by a significant decrease in glutathione (GSH) and the activities of glutathione peroxidase (GPx), glutathione-S-transferase (GST) and gamma glutamyl transpeptidase (GGT). Administration of garlic and neem leaf extracts significantly decreased the formation of lipid peroxides and enhanced the levels of antioxidants and detoxifying enzymes in stomach, the primary target organ for MNNG, as well as in the liver and circulation. The results of the present study suggest that garlic and neem may exert their protective effects by modulating lipid peroxidation and enhancing the levels of GSH and GSH-dependent enzymes. PMID- 10686580 TI - Angiotensin converting enzyme (ACE) activity levels in insulin-independent diabetes mellitus and effect of ACE levels on diabetic patients with nephropathy. AB - Involvement of complications is considered to be one of the major factors in the prognosis of diabetes mellitus (DM). Recent studies indicate that most diabetic complications such as nephropathy and hypertension are vascular-originated. Renin angiotensin involvement, especially changes in ACE activity level, is considered to be a key factor since ACE converts angiotensin I to angiotensin II which is a potent vasoconstrictor and plays a vital role in the regulation of blood pressure. Our present study focused on ACE activity levels along with blood glucose and HbA(1c) levels in diabetic patients with (n=18) or without (n=25) nephropathy as compared to control subjects (n=25). Blood glucose levels were significantly higher in both diabetic groups compared to controls (p<0.001). On the other hand, compared to controls, blood HbA(1c) levels were slightly higher in DM patients without complications whereas they were significantly increased in nephropatic DM patients (p<0.001). There was a very strong increase (p<0.001) at the level of ACE activity in both of the diabetic groups (with nephropathy: 47.11+/-3.70 U l(-1); without complications: 43.72+/-2.93 U l(-1); controls: 25.15+/-2.30 U l(-1)). ACE activity levels were also significantly higher in diabetic patients with nephropathy than in type II DM patients without complication (p<0.01). Our results demonstrate that ACE activity levels are increased in diabetic patients. Additional significant increase in ACE activity levels in diabetic patients with complications such as nephropathy supports the hypothesis that ACE activity has an essential role in the development of complications in diabetes. PMID- 10686581 TI - Intracellular protein degradation and autophagy in isolated pancreatic acini of the rat. AB - Simultaneous investigation of protein degradation and autophagy of isolated exocrine pancreatic cells is carried out here for the first time in a systematic way by a complex biochemical, morphological and morphometrical approach. Protein degradation proceeds with a decreasing rate of 4-1.5 per cent per h over a 4-h period indicating a comparatively low degradation capacity. Cells in freshly isolated acini do not contain autophagic vacuoles but the latter appear within an hour in vitro and their quantity remains close to a steady state during the subsequent 3 h. Both traditional inhibitors of the autophagic-lysosomal pathway, e.g. vinblastine, leupeptin, and lysosomotropic amines together with the recently introduced 3-methyladenine, inhibit degradation to a similar maximal extent, offering the possibility of the estimation of the ratio of lysosomal/non lysosomal degradation. In pancreatic acinar cells autophagic sequestration is unaffected and protein degradation is inhibited inside secondary lysosomes by leupeptin and lysosomotropic amines, while 3-methyladenine prevents the formation of autophagosomes. Vinblastine seems to act by inhibiting the fusion of autophagosomes with lysosomes and there is no evidence for the stimulation of autophagic sequestration by vinblastine in the present system. The effect of inhibitors of protein breakdown on protein synthesis is variable and does not correlate with their influence on degradation. Amino acids strongly stimulate protein synthesis, but in contrast to what is found in liver cells, they do not seem to affect protein degradation or autophagy significantly, thus indicating major regulatory differences of these processes between pancreatic acinar cells and hepatocytes. PMID- 10686582 TI - Age-related changes in GM1, GD1a, GT1b components of gangliosides in Wistar albino rats. AB - In this study, age-related changes of GM1, GD1a, GT1b fractions of gangliosides were investigated in whole brain of male Wistar albino rats. Insignificant increases were detected in GM1 values from the third to the 24th month, whereas GD1a and GT1b concentrations of ganglioside in 24-month-old rats decreased significantly as compared to 6-month-old rats. Although there were no significant differences in the GD1a/GT1b ratio of any groups, GM1/GD1a and GM1/GT1b ratios were significantly increased as compared to 6-month-old rats. The increase in the ratios of gangliosides are not due to an increase of GM1 fractions; they result from a decrease of GD1a and GT1b fractions of gangliosides. In conclusion, the concentration of ganglioside decreased with ageing. PMID- 10686583 TI - Extracellular release of free fatty acids by rat T lymphocytes is stimulus dependent and is affected by dietary lipid manipulation. AB - [(3)H]-Arachidonic acid-labelled rat T lymphocytes released radioactivity extracellularly when stimulated by the calcium ionophore A23187 or by monoclonal antibodies to some cell surface structures (CD2, CD5, CD11a, CD18, CD54, T-cell receptor) but not to others (CD49d, CD62L); release was greater with the calcium ionophore. Almost all of the radioactivity released from anti-CD2-stimulated lymphocytes was recovered in the free fatty acid fraction, whereas only about 50 per cent of that released after A23187 stimulation was recovered in this fraction. A23187 stimulation resulted in release of arachidonic acid from a variety of phospholipids (phosphatidylinositol, phosphatidylcholine and perhaps phosphatidylethanolamine), while the monoclonal antibody stimulation released arachidonic acid from phosphatidylinositol and perhaps phosphatidylcholine. Unstimulated lymphocytes released a range of fatty acids extracellularly, with palmitic acid accounting for 35-40 per cent and arachidonic acid for 5 per cent of released fatty acid. Stimulation of lymphocytes with either anti-CD2 or A23187 increased total fatty acid release 1.5- to 1.8-fold. In both cases palmitic acid remained the most predominant fatty acid released but the contribution of arachidonic acid increased. The type of lipid fed to the rats significantly influenced the amount and type of fatty acid released. Fish oil feeding significantly reduced extracellular fatty acid release by stimulated lymphocytes. PMID- 10686584 TI - Elevated levels of Ca(II) modulate the activity and inhibition of serine proteases: implication in the mechanism of apoptosis. AB - Elevated levels of intracellular Ca(II) are a prominent feature of apoptosis, a natural form of cell death involved in many physiological and pathological processes. Serine proteases play crucial roles in apoptosis and have been implicated in the genomic DNA degradation and the massive protein degradation that occur during apoptosis. In this study, the effects of the elevated level of Ca(II) on the activity and inhibition of serine proteases were examined by spectrophotometric methods. The effects of the elevated levels of Ca(II), Mg(II), K(I), and Na(I) on the activity and inactivation of three representative members of serine proteases were determined. The level of serine protease activity in CEM C7-14 leukemic cells was also evaluated in the presence and absence of dexamethasone-induced apoptosis, and also in the presence of A23187, a Ca(II) ionophore. Among the four metal-ions studied, only Ca(II) was found to significantly enhance the activity of mammalian serine proteases. Ca(II) was also found to significantly protect the enzymes from inhibition, while the other three metal-ions showed no significant effect on the inactivation of the enzymes. Compared to the control sample, the enzymic activity was found to be higher during apoptosis, and in the presence of the Ca(II)-ionophore. Results of this study indicate that Ca(II) can significantly enhance the catalytic efficiency of serine proteases during apoptosis. PMID- 10686585 TI - Extracellular Ca(2+) suppresses endotoxin-inducible tissue factor activation in monocytic THP-1 cells. AB - BACKGROUND: Monocytic tissue factor (TF), an initiator of extrinsic blood coagulation, is often activated under various inflammatory conditions including endotoxemia. This activation could be a contributing factor to the manifestation of disseminated intravascular coagulation following septic shock. HYPOTHESIS: We herein determine if extracellular Ca(2+) ([Ca(2+)](ex)) regulates bacterial endotoxin (LPS)-inducible monocytic TF activation. METHODS: We have employed a model monocytic cell line (THP-1) to explore the mode of action of [Ca(2+)](ex) on the modulation of LPS-induced TF activation. TF activity was measured by a single stage clotting assay, while TF expression as well as LPS recognition and its receptor expression were studied in immunofluorescent approaches. RESULTS: LPS-induced TF activation was inversely correlated to [Ca(2+)](ex). Upon exposure of THP-1 cells to LPS (1.5 microg ml(-1)) for 6 h in the Hanks' medium without CaCl(2), TF was activated by nearly 10-fold. TF activation appreciably decreased with the increasing [Ca(2+)](ex). No more than 3.5-fold TF activation was detected at 5 mM [Ca(2+)](ex). Consistent with the significantly lower degree of TF activation, LPS-induced TF expression at 5 mM [Ca(2+)](ex) was 60 per cent less than that without [Ca(2+)](ex). FACScan analysis showed that LPS recognition was significantly blocked at 5 mM [Ca(2+)](ex) which however had no effect on the expression of CD14 and CD11b, the proposed major LPS receptors. Moreover, LPS binding in vitro was significantly inhibited by 5 mM CaCl(2). CONCLUSION: Our results demonstrate that [Ca(2+)](ex) blocked LPS recognition without affecting its receptor expression on THP-1 monocytes. This insensitivity to LPS thereby resulted in the depressed inducible monocytic TF expression and activation. PMID- 10686586 TI - Transient expression of the glial glutamate transporters GLAST and GLT in hippocampal neurons in primary culture. AB - The extracellular glutamate concentration is kept low by glutamate transporters in the plasma membranes. Here we have studied the expression of the glutamate transporters GLAST, GLT and EAAC during the in vitro development of embryonic hippocampal neurons grown in a defined (serum free) medium. Immunochemistry studies showed that both the GLAST and GLT proteins are expressed in a subpopulation of neurons at the early, but not at the later stages of the cultures. Glial cells expressing the GLAST and GLT proteins were found at all stages. EAAC was only detected in neurons. This is one of the first evidence of a neuronal ability to express GLAST. PMID- 10686587 TI - Activation of CA(2+)/calmodulin-dependent protein kinase IV in cultured rat hippocampal neurons. AB - Ca(2+)/calmodulin-dependent protein kinase IV (CaM kinase IV) is a multifunctional enzyme that is abundantly present in the nuclei of neurons. We report the properties of phosphorylation and activation of CaM kinase IV in comparison to CaM kinase II in cultured rat hippocampal neurons. Phosphorylation and activity of CaM kinase IV as well as CaM kinase II were increased by treatment of neurons either with glutamate or high K(+). Glutamate-induced phosphorylation and activity of CaM kinase IV were blocked by N-methyl-D-asparate (NMDA) antagonists, and NMDA application instead of glutamate did increase CaM kinase IV phosphorylation. CaM kinase IV phosphorylation was also increased by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), and was blocked by an inhibitor of NMDA receptor. The AMPA-induced phosphorylation was blocked by tetrodotoxin, a Na(+) channel blocker, that was expected to block endogenous glutamate transmission indirectly. On the other hand, high K(+)-induced phosphorylation and activation were not blocked by inhibitors of glutamate receptors, and effectively blocked by nifedipine, an L-type Ca(2+) channel blocker. These properties were similar between CaM kinase IV and CaM kinase II. PMID- 10686588 TI - Lactacystin, a specific inhibitor of the proteasome, induces apoptosis and activates caspase-3 in cultured cerebellar granule cells. AB - The multicatalytic protease complex or proteasome is a fundamental nonlysosomal tool that the cell uses to process or degrade proteins at a fast rate through the ubiquitin and ATP-dependent proteolytic pathway. Examples of these important proteins include the tumor suppressor protein p53, various cyclins, the cyclin dependent kinase inhibitor p27, NFkappaB, IkappaB, c-fos, and c-jun. The activation of proteolytic enzymes, including certain cystein-proteases of the ced 3/ICE (interleukin-1beta-converting enzyme) family, is a characteristic feature of the apoptotic program. However, the role of the multicatalytic protease complex in apoptosis is not well known. In order to obtain further information regarding the participation of the ubiquitin-mediated pathway in the decision of the cell to execute the cell death program, we have used a specific inhibitor of the multicatalytic protease complex, lactacystin, in cultured cerebellar granule cells. Cells were obtained from the cerebellum of 6- to 8-day-old Wistar rats and cultured in Neurobasal medium supplemented with B-27. Addition of lactacystin to the cultures induced apoptosis of the granule cells in a time-dependent fashion. The morphological changes produced by the proteasome inhibitor included nuclear condensation and DNA fragmentation measured by the diphenylamine test, as well as a positive labeling by the TUNEL (terminal deoxynucleotidyltransferase mediated dUTP nick end labeling) assay, all of them typical features of apoptosis. Concomitant with apoptosis, there were changes in the expression of the ubiquitin mRNA, a progressive depletion in the free ubiquitin pool, and an increase in the high molecular weight ubiquitin-protein conjugates. Caspase-3, a member of the ced-3/ICE family of cystein-proteases, showed a marked increase in activity in the lactacystin-treated cells. In flow cytometry studies, the amount of cells in the S phase of the cell cycle was smaller in the lactacystin-treated cells than in controls, suggesting that apoptosis could be due, in part, to an alteration of the cell cycle. PMID- 10686589 TI - Roles of neuregulin in synaptogenesis between mossy fibers and cerebellar granule cells. AB - Neuregulins (NRGs), a large group of structurally related signaling proteins, are likely to have important roles in the development, maintenance and repair of the nervous system and other selected tissues. We have demonstrated, by using the major form of NRG cloned from the mouse cerebellum that both the soluble form and the membrane anchored form of NRG may serve different functions in synaptogenesis. The soluble form of NRG was produced by proteolytic cleavage of the membrane anchored form of NRG. The proteolytic cleavage was promoted by protein kinase activation. The cleaved form of NRG trans-synaptically regulated the expression of the NMDA (N-methyl-D-aspartate) receptor subunit NR2C as neurally-derived factors, whereas the membrane anchored form of NRG showed a homophilic binding activity between NRGbeta1s. In adult mice the membrane anchored form of NRG was concentrated in neuro-terminals of both granule cells and pontocerebellar mossy fibers. The fact that NRG can be functionally viewed as cell recognition molecules as well as neurotrophic agents suggests new possibilities for the important class of molecules. PMID- 10686590 TI - Cyclic AMP regulates substance P expression in developing and mature spinal sensory neurons. AB - The tachykinin, substance P, has long been associated with transmission of noxious stimuli. However, relatively little is known about signal transduction pathways subserving peptidergic regulation in sensory neurons. To investigate whether cyclic AMP (cAMP) could be a potential second messenger subserving substance P expression, dorsal root ganglion neurons were grown in culture in the presence of agents that increase content of cAMP. In developing neurons, forskolin increased substance P content and survival almost threefold. Anti-nerve growth factor (NGF) blocked the effect of NGF but not forskolin, suggesting that increased cAMP acts directly and not via increased secretion of NGF from Schwann cells and fibroblasts. In adult neurons, which do not require supplemental trophic factors for survival, NGF and forskolin had similar effects on substance P levels. Neither agent had any effect on somatostatin content of either developing or mature sensory neurons. 8-bromo cAMP and isobutyl methylxanthine duplicated the action of forskolin. Further, all three agents increased expression of preprotachykinin mRNA. Forskolin appeared to increase both total and neuron-specific expression of message as well as the number of neurons expressing mRNA. Our results suggest that cAMP directly regulates substance P content in sensory neurons from adult and neonatal rats. PMID- 10686591 TI - Balance of two secretion pathways of nerve growth factor in PC12 cells changes during the progression of their differentiation, with a decrease in constitutive secretion in more differentiated cells. AB - Proteins are secreted from animal cells by either a constitutive or a regulated pathway. When cDNA of nerve growth factor (NGF) was introduced into PC12 cells, these cells produced and secreted active NGF, where NGF was secreted not only in constitutive but also in activity-dependent regulated way according to the results of pulse-chase and ELISA studies. The regulated secretion was caused by depolarization, cyclic AMP analogue, or beta-adrenergic agonist but not by glutamate or carbachol. Because these transfected cells differentiated into a morphology indistinguishable from that incubated with NGF protein, we next compared the secretion pathways of NGF from PC12 cells at different stages of the differentiation. NGF was secreted in both constitutive and regulated way at 2 and 7 days after the transfection of NGF-cDNA, but the constitutive secretion of NGF from the more differentiated cells of Day 7 was decreased and mature NGF tended to accumulate in the cells. These results indicate that the neurotrophin secretion mechanism is intimately regulated in the course of the differentiation of PC12 cells. Such a change in the protein secretion pathway might have an profound role in the development of neurons. PMID- 10686592 TI - Meizothrombin, an intermediate of prothrombin activation, stimulates human glioblastoma cells by interaction with PAR-1-type thrombin receptors. AB - Thrombin induces well-characterized effects on normal and neoplastic brain cells by interaction with protease-activated receptor (PAR)-type thrombin receptors. However, nothing is known about the function of intermediate enzymes of prothrombin activation recently shown to evoke PAR-1-mediated signaling in smooth muscle cells. Therefore, we investigated the effect of recombinant human meizothrombin (rMT), one of thrombin's catalytically active precursor enzymes in the prothrombin cleavage cascade, on calcium mobilization in human SNB-19 glioblastoma cells. By using reverse-transcription polymerase chain reaction, immunofluorescence studies with a monoclonal anti-PAR-1 antibody and calcium measurements, SNB-19 cells were shown to express functional PAR-1-type thrombin receptors. PAR-1 is not only a receptor for thrombin in SNB-19 cells but was also activated by rMT very effectively. Under the conditions used in our experiments, SNB-19 cells stimulated with thrombin after rMT challenge were unable to elicit a new calcium response and vice versa. In addition, both rMT and thrombin induced no further calcium signal after that observed with the PAR-1-activating peptide SFLLRN. Therefore, rMT and thrombin seem to activate calcium signaling by similar mechanisms including PAR-1. Our results demonstrate rMT as a potent activator of PAR-1-type thrombin receptors in SNB-19 glioblastoma cells, suggesting a function of catalytically active thrombin precursor enzymes in cells of glial origin. PMID- 10686593 TI - Uptake of circulating insulin-like growth factor-I into the cerebrospinal fluid of normal and diabetic rats and normalization of IGF-II mRNA content in diabetic rat brain. AB - Brain injury has been prevented recently by systemic administration of human insulin-like growth factor-I (hIGF-I). It is widely believed that protein neurotrophic factors do not enter the brain from blood, and the mechanism by which circulating hIGF-I may be neuroprotective is uncertain. This investigation tested the hypothesis that hIGF-I is taken up into cerebrospinal fluid (CSF) from the circulation. (125)I-hIGF-I was injected subcutaneously into rats. The (125)I IGF-I recovered from CSF and plasma were indistinguishable in size from authentic (125)I-hIGF-I on SDS-PAGE. An ELISA was used that detected immunoreactive hIGF-I, but not rat IGF-I, rat IGF-II, human IGF-II, or insulin. Osmotic minipumps were implanted for constant subcutaneous infusion of various hIGF-I doses. Uptake into CSF reached a plateau at plasma concentrations above approximately 150 ng/ml hIGF I; the plateau was consistent with carrier-mediated uptake. The plasma, but not CSF, hIGF-I level was significantly reduced in streptozotocin diabetic vs. nondiabetic rats, and uptake of hIGF-I into CSF was nonlinear with respect to plasma hIGF-I concentrations. Nonlinear uptake excluded leakage or transmembrane diffusion of IGF-I from blood into CSF as a dominant route for entry, but the site and mechanism of uptake remain to be established. The IGF-II mRNA content per milligram brain (P < 0.02) as well as per poly(A)(+) RNA (P < 0.05) was significantly increased towards normal in diabetic rats treated by subcutaneous administration of hIGF-I vs. vehicle. This effect of circulating hIGF-I may have been due to regulation of IGF-II gene expression in the choroid plexus and leptomeninges, structures at least in part outside of the blood-central nervous system barrier. These data support the hypothesis that circulating IGF-I supports the brain indirectly through regulation of IGF-II gene expression as well as by uptake into the CSF. PMID- 10686595 TI - Fibroblast growth factor modulates HIV coreceptor CXCR4 expression by neural cells. HNRC Group. AB - Recent studies suggest that the chemokine receptor CXCR4 may be involved in mediating the neurodegenerative process in the brains of patients with acquired immunodeficiency disease (AIDS). In this context, we hypothesize that neurotrophic factors, such as fibroblast growth factor (FGF), might protect against human immunodeficiency virus (HIV)-mediated neurotoxicity via regulating the expression of CXCR4 in neural cells. For this purpose, levels of CXCR4 were determined in neuronal and glial cell lines after FGF1 and 2 treatment. In addition, levels of CXCR4 immunoreactivity were associated with levels of FGF1 immunoreactivity in the brains of HIV-positive patients. These studies showed that neuronal CXCR4 levels decreased in a dose-dependent manner after exposure to FGF. Conversely, glial CXCR4 was increased in a dose-dependent manner after FGF2 treatment. These effects were dependent on the FGF receptor tyrosine kinase signaling pathway, because FGF-induced effects on CXCR4 were blocked by the tyrosine kinase inhibitor, 5'-deoxy-5'methylthioadenosine, or by anti-FGF receptor antibody. Stromal cell-derived factor-1, the ligand for CXCR4, and HIV gp120 neurotoxicity was attenuated by FGF1 in a dose-dependent manner in vitro, further supporting physiological relevance. In the brains of AIDS patients, the levels of neural CXCR4 immunoreactivity were inversely associated with FGF levels. Taken together, these results support the possibility that the neuroactive effects of FGF in HIV encephalitis might be mediated through regulation of the expression of CXCR4. PMID- 10686594 TI - Short-term treatment with interferon-alpha/beta promotes remyelination, whereas long-term treatment aggravates demyelination in a murine model of multiple sclerosis. AB - The mechanisms by which type I interferons (IFN) reduce the rate and severity of exacerbations in multiple sclerosis are unknown. We utilized a model of multiple sclerosis to determine the extent of demyelination and remyelination in Theiler's murine encephalomyelitis virus (TMEV)-infected SJL/J mice treated with mouse IFN alpha/beta for a short (5 weeks) or a long (16 weeks) period. All mice were chronically infected with TMEV to simulate the clinical situation in multiple sclerosis. Short-term IFN-alpha/beta treatment increased the percent of remyelinated spinal cord white matter by threefold when compared with phosphate buffered saline (PBS) treatment (P < 0.02), but it did not affect the extent of demyelination. In contrast, long-term IFN-alpha/beta treatment increased the extent of demyelination by twofold (P < 0.03). Long-term treatment increased the absolute area of remyelination, but the percent remyelination as a function of area of demyelination was not changed because of increased demyelination. An immunomodulatory mechanism may have contributed to the effect of IFN-alpha/beta on white matter pathology because treated mice had higher anti-TMEV IgGs in serum and demonstrated decreased numbers of B and T lymphocytes infiltrating the central nervous system (CNS). There was no correlation between the level of anti- IFN-alpha/beta antibodies and the extent of demyelination or remyelination. These results indicate that the length of type I IFN treatment may have paradoxical effects on demyelination and remyelination. PMID- 10686596 TI - Altered cell-matrix associated ADAM proteins in Alzheimer disease. AB - Alterations in cell-matrix 'contact' are often related to a disruption of cell cycle regulation and, as such, occur variously in neoplasia. Given the recent findings showing cell cycle alterations in Alzheimer disease, we undertook a study of ADAM-1 and 2 (A Disintegrin And Metalloprotease), developmentally regulated, integrin-binding, membrane-bound metalloproteases. Our results show that whereas ADAM-1 and 2 are found in susceptible hippocampal neurons in Alzheimer disease, these proteins were not generally increased in similar neuronal populations in younger or age-matched controls except in association with age-related neurofibrillary alterations. This increase in both ADAM-1 and 2 in cases of Alzheimer disease was verified by immunoblot analysis (P < 0.05). An ADAM-induced loss of matrix integration would effectively "reset" the mitotic clock and thereby stimulate re-entry into the cell cycle in neurons in Alzheimer disease. Furthermore, given the importance of integrins in maintaining short-term memory, alterations in ADAM proteins or their proteolytic activity could also play a proximal role in the clinico-pathological manifestations of Alzheimer disease. PMID- 10686597 TI - L-lactate inhibits L-cystine/L-glutamate exchange transport and decreases glutathione content in rat cultured astrocytes. AB - In several brain pathologies, the level of brain L-lactate increases. The stimulation of L-lactate production is a detrimental factor in promoting neuronal cell damage and astrocytic dysfunction. Astrocytic glutathione metabolism has an important role to protect brain cells against oxidative stress. In this study, effects of L-lactate on L-cystine uptake and glutathione level in rat-cultured astrocytes were examined. L-Lactate decreased the L-(35)S-cystine and Na(+) independent L-(3)H-glutamate uptakes into astrocytes at the concentrations more than 2.5 mM. The L-lactate-induced decrease in L-(35)S-cystine uptake was neither affected by modification of extracellular pH nor mimicked by acetate, propionate and butyrate. The apparent Km value of the L-(35)S-cystine uptake was increased by L-lactate, while the Vmax was not changed. Astrocytic glutathione and nonprotein thiol content was decreased by incubation with 20 mM L-lactate for 48 hours (65% and 75% of control values, respectively). The decreases in astrocytic glutathione and nonprotein thiol content were restored to normal levels by withdrawal of L-lactate. These results suggest that L-lactate inhibits astrocytic L-cystine/L-glutamate exchangers and affects the glutathione contents. PMID- 10686598 TI - Fiber types in the mouse levator auris longus muscle: a convenient preparation to study muscle and nerve plasticity. AB - The histochemical composition of the levator auris longus (LAL) muscle has been investigated in adult NMRi mice. Histochemical reaction for myofibrillar adenosine triphosphatase (ATPase) after preincubation in alkaline and acidic media, nicotine amideadenine-dinucleotide dehidrogenase (NADH-dehydrogenase), and alpha-glycerophosphate dehydrogenase were performed on cryosections of LAL muscle. Expression of myosin heavy chain (MyHC) isoforms was detected with the immunoperoxidase method applying monoclonal antibodies against MyHC isoforms -1, 2a, -2x/d, and -2b, as well as by sodium dodecylsulfate (SDS) glycerol gel electrophoresis. The muscle was proven to be a pure fast-twitch muscle. The most numerous fibers in LAL muscles contained MyHC-2b and some MyHC-2a. Histochemically, pure IIA fibers with oxidative metabolism and pure IIB fibers with glycolytic metabolism were detected. In contrast to the majority of mature control muscles, numerous hybrid fibers coexpressing MyHC-2x/d with MyHC-2a or MyHC-2b were present. Both hybrids were oxidative-glycolytic; additionally, some hybrids containing MyHC-2a were oxidative. In one out of six muscles, traces of MyHC-1 were detected both with immunoperoxidase staining and with SDS glycerol gel electrophoresis. Rare fibers that exceptionally expressed small amounts of MyHC-1 always coexpressed MyHC-2a, which is an additional proof that pure type I fibers do not exist in LAL. Due to these histochemical characteristics and to its previously described morphological features, the use of the LAL muscle as a model for various studies, particularly muscle and nerve interactions, is emphasized. PMID- 10686599 TI - Cre recombinase: the universal reagent for genome tailoring. PMID- 10686600 TI - Expression of Cre recombinase in mouse oocytes: a means to study maternal effect genes. PMID- 10686601 TI - Epiblast-restricted Cre expression in MORE mice: a tool to distinguish embryonic vs. extra-embryonic gene function. PMID- 10686602 TI - Targeted insertion of Cre recombinase into the TNAP gene: excision in primordial germ cells. PMID- 10686603 TI - HoxB6-Cre transgenic mice express Cre recombinase in extra-embryonic mesoderm, in lateral plate and limb mesoderm and at the midbrain/hindbrain junction. PMID- 10686604 TI - Cre recombinase expression in the floorplate, notochord and gut epithelium in transgenic embryos driven by the Hoxa-1 enhancer III. PMID- 10686605 TI - Expression pattern of a Krox-20/Cre knock-in allele in the developing hindbrain, bones, and peripheral nervous system. PMID- 10686606 TI - Targeted expression of Cre recombinase to myelinating cells of the central nervous system in transgenic mice. PMID- 10686607 TI - Retina- and ventral forebrain-specific Cre recombinase activity in transgenic mice. PMID- 10686608 TI - CaMKIIalpha-Cre transgene expression and recombination patterns in the mouse brain. PMID- 10686609 TI - Muscle specific expression of Cre recombinase under two actin promoters in transgenic mice. PMID- 10686610 TI - Analysis of the Cre-mediated recombination driven by rat insulin promoter in embryonic and adult mouse pancreas. PMID- 10686611 TI - Mosaic Cre-mediated recombination in pancreas using the pdx-1 enhancer/promoter. PMID- 10686612 TI - Col2a1-directed expression of Cre recombinase in differentiating chondrocytes in transgenic mice. PMID- 10686613 TI - Inducible site-specific somatic mutagenesis in mouse hepatocytes. PMID- 10686614 TI - DNA excision in liver by an albumin-Cre transgene occurs progressively with age. PMID- 10686615 TI - Hepatocyte-specific expression of Cre recombinase. PMID- 10686616 TI - Prostate specific expression of Cre recombinase in transgenic mice. PMID- 10686617 TI - Ubiquitous postnatal LoxP recombination using a doxycycline auto-inducible Cre transgene (DAI-Cre). PMID- 10686618 TI - Characterization of an inducible, epidermal-specific knockout system: differential expression of lacZ in different Cre reporter mouse strains. PMID- 10686619 TI - Neural crest expression of Cre recombinase directed by the proximal Pax3 promoter in transgenic mice. PMID- 10686620 TI - Selective expression of Cre recombinase in skeletal muscle fibers. PMID- 10686621 TI - Grip form and graphomotor control in preschool children. AB - OBJECTIVE: The purpose of this study was to examine the utility of the grip scale presented by Schneck and Henderson, the effect of grip form on drawing accuracy, and the effect of implement diameter on grip form and drawing accuracy. METHOD: Sixty boys and girls who were 3, 4, and 5 years of age performed 20 trials of a precision drawing task, 4 trials each with five implements of varying diameters (4.7, 7.9, 11.1, 14.3, and 17.5 mm). RESULTS: First, all 1,200 grips could be coded according to Schneck and Henderson's 10-grip whole-configuration assessment system, but the interrater reliability was lower than expected (.67 proportion of perfect agreement). Second, using Schneck's five-level scoring system, the level of grip significantly affected drawing accuracy, with the highest grip level used most often with the highest accuracy scores and the lowest observed grip level used most often with the lowest accuracy scores. Third, increasing implement diameter led to significantly lower level grips but did not significantly affect accuracy. CONCLUSIONS: Therapists are recommended to use Schneck and Henderson's 10-grip scale only for documenting the persons' grips and changes in their grips, but if comparisons between individual persons are desired, then Schneck's five level scale, which affords greater generalizability, should be used. Further, children with graphomotor performance deficits are not likely to benefit from grip manipulations because such strategies were shown to make better only performance that is already good. PMID- 10686622 TI - Clinical interpretation of "grip form and graphomotor control in preschool children". PMID- 10686623 TI - Developing a context-based performance measure for persons with schizophrenia: the test of grocery shopping skills. AB - OBJECTIVE: This article describes how authenticity and directness, desired characteristics of performance measures, were applied to the development of a context-based Test of Grocery Shopping Skills (TOGSS) for persons with schizophrenia. METHOD: The steps used in developing the measure included interviewing consumers with schizophrenia to identify issues in grocery shopping, conceptualizing how authenticity and directness could be applied to shopping performance, and selecting grocery items to be used in the test. The two forms of the TOGSS were administered to 26 persons with schizophrenia or schizoaffective disorders to evaluate reliability (stability, equivalence, interrater) and validity (convergent, generalizability) of the TOGSS. RESULTS: The correlations between the two forms of the TOGSS over two different testing periods were significant, ranging from .64 to .83. Subscale scores were moderately correlated (R = .52 to .94) with a similarly constructed test of drugstore shopping. CONCLUSION: A systematic method can be used to develop a context measure of performance. The TOGSS has beginning evidence of reliability and validity. With further study, the test will be useful in assessing the independent living skill of grocery shopping in persons with psychiatric disorders. PMID- 10686624 TI - Understanding the experience of noninclusive occupational therapy clinics: lesbians' perspectives. AB - This article presents understandings about how noninclusive occupational therapy environments are developed and maintained. The data are drawn from a study that, in part, explored the experiences of lesbian or bisexual occupational therapists working in a health care system. Ten participants each engaged in two to five in depth audiotaped interviews. The narrative data were analyzed with a modified form of grounded theory. The data provide insight into how heterosexist occupational therapy work climates are created and maintained through four processes: heterosexual discourse, homophobic comments, assumed heterosexuality, and perceived stereotypes. The way in which heterosexist occupational therapy work climates may impede the professional growth of therapists also is presented. The knowledge gained from this study can help practitioners, professors, and students in their attempts to sustain an inclusive environment with respect to persons who are lesbian, gay, and bisexual. PMID- 10686625 TI - Parental hopes for therapy outcomes: children with sensory modulation disorders. AB - OBJECTIVE: Understanding parents' hopes for therapy outcomes is essential to family-centered care. This qualitative study explored parents' points of view regarding their hopes for the outcomes of occupational therapy using a sensory integration treatment approach. METHOD: Data were collected as part of a larger research project on the effectiveness of rehabilitating children who have sensory modulation disorders. Five interviews were randomly selected from 17 parent interviews conducted in the larger study. Data were analyzed using grounded theory methods. FINDINGS: Three themes pertinent to the occupations of children and two themes related to the occupations of parenting and sustaining family life emerged. Child-focused outcomes include social participation, self-regulation, and perceived competence. Parent-focused outcomes include learning strategies to support children and obtaining personal validation. DISCUSSION: Interventions are proposed that relate to children's participation in contexts in which they live, learn, and play, as well as the support of parents in the occupations of parenting. PMID- 10686626 TI - Assessing father-infant interactions using the NCAST teaching scale: a pilot study. AB - OBJECTIVE: The purpose of this pilot study was to gather preliminary data on father-infant dyads using the Nursing Child Assessment Satellite Training (NCAST) Teaching scale, a parent-infant interaction measure, to determine whether and how fathers score differently than mothers from normative samples. METHOD: Interactions between first-time father (N = 15) and their infants, 3 months to 6 months of age, during the instruction of an unfamiliar play activity were rated using the NCAST Teaching scale. Scores were compared both with a normative database (N = 2,123) of mother-infant dyads and with a subsample (n = 34) of the normative database to control for demographic variables, including the age, gender, and birth parity of the child and the age, education, marital status, and ethnicity of the parent. RESULTS: The fathers scored significantly lower on items related to fostering the infants' cognitive growth than the mothers in the normative database. However, the infants in this study provided clearer behavioral cues and were more responsive to their fathers than the infants in the normative sample. These findings were also true for the subsample comparison. The fathers also scored significantly lower than the normative subsample on items measuring their ability to foster the social and emotional growth of their infants. CONCLUSION: There may be important differences in the interactions of father-infant dyads compared with mother-infant dyads, but further research with a larger, more representative sample of fathers on this parent-infant interaction measure is warranted to support this. The development of normative scores for fathers and their infants is recommended to accurately interpret father-infant interactions when administering the NCAST Teaching scale. PMID- 10686627 TI - Mothers' perceptions of child care assistance: the impact of a child's disability. AB - OBJECTIVE: This study examined and compared mothers' perceptions of child care assistance provided by fathers and other caregivers. Awareness of child care division of labor will assist occupational therapists in addressing the needs of children with disabilities within the family context. METHOD: One hundred and thirty-five mothers living in two-parent households kept a time diary of their daily activities for 7 consecutive days using the Caregiver's Activity and Recording of Events Inventory and estimated the percentage of child care their partners performed, the amount of child care their partners performed, and their satisfaction with this division of labor. One third of the women had children with multiple disabilities, one third had children with Down syndrome, and one third had children who were typically developing. RESULTS: The majority of mothers in all three groups perceived that they were responsible for the majority of child care. There were no significant differences between groups in terms of mothers' perceptions of the amount of child care provided by fathers and other caregivers, including relatives, childsitters, nurses, school personnel, and neighbors. However, there were wide variations among families concerning child care arrangements and division of labor. Seventy-five percent of mothers indicated that they were satisfied with the division of child care labor between mothers and fathers, and no significant correlation was found between perceived percentage of child care performed and satisfaction with the division of labor. CONCLUSION: Mothers in this study were responsible for the majority of child care whether their child had a disability. The variation in number of hours that others spent performing child care activities within individual families suggests that there is no "best" or typical pattern. Occupational therapists need to collaborate with families to determine a system of accommodations to manage their daily routine that most effectively meets the family's needs. PMID- 10686628 TI - Current parent education on infant feeding in the neonatal intensive care unit: the role of the occupational therapist. AB - OBJECTIVES: The purpose of this study was to describe current trends in parent education on infant feeding in the neonatal intensive care unit (NICU) and to clarify the role of the occupational therapist in educating parents. METHOD: Questionnaires were mailed to 190 neonatologists across the United States who were asked to forward it to a NICU occupational therapist. The questionnaire gathered descriptive information about the structure of parent education in the NICU, the role of the occupational therapist in providing parent education, and demographics about respondents and their NICUs. The response rate was 53% (n = 100). RESULTS: All 100 hospitals responding provided parent education in some form, and most included a variety of topics and teaching methods. Occupational therapists were on the NICU team at 74 of the hospitals and were identified third most frequently as a provider of parent education. The occupational therapists were most frequently identified as responsible for teaching about positioning, infant development, and infant states and cues and were highly involved in educating parents about feeding. CONCLUSION: Current parent education programs in NICUs are comprehensive in scope. Occupational therapists' role in educating parents about infant care and feeding consists of a focus on certain topics where occupational therapists have specialized skills and education. Occupational therapists are recognized by their NICU colleagues as providers of parent education, but this study suggests that the occupational therapists' role may not be clearly understood by other NICU professionals. PMID- 10686629 TI - Dynamic performance analysis: a framework for understanding occupational performance. AB - Occupational therapy is now consistently described as a profession concerned with enabling occupation. A crucial step in enabling occupation is understanding the occupational performance of our clients. Dynamic Performance Analysis (DPA) is a new approach to occupational analysis that focuses on the client's actual performance. DPA, acknowledging that optimal performance is the product of the interaction of person, environment, and occupation, and thus highly individualistic, places the client and his or her occupation, in interaction with the environment, at the center of the analysis process. Embedded in a top-down framework, DPA is a dynamic, iterative process, carried out as the client performs the occupation. The purpose of DPA is to identify where performance breaks down and test out solutions. In this article, the rationale, origins, and basic assumptions of DPA are discussed, and a detailed description of the DPA process together with two clinical examples is presented. PMID- 10686630 TI - Playfulness in children with and without disability: measurement and intervention. AB - OBJECTIVE: The differences in playfulness between young children with cerebral palsy and developmental delays and children who are typically developing, and the comparative effects of two interventions (one focused on improving mother-child interaction patterns, the other a neurodevelopmental treatment [NDT] session) on children's playfulness were examined in this study. Reliability and validity of the Test of Playfulness (ToP) also were examined. METHOD: Three trained raters used the ToP to score 38 children, half with cerebral palsy and developmental delays and half typically developing, as they played with their mothers. Mental ages of the children ranged from 3 to 18 months. The mother-child dyads in which the children had cerebral palsy and developmental delays were then randomly assigned to an intervention group. After a 1-hr intervention to improve mother child interaction, the children were rescored on the ToP. RESULTS: After examination of ToP reliability and validity, children with cerebral palsy and developmental delays were found to score significantly lower on the ToP than their peers who were typically developing. In addition, children whose mothers received an intervention to improve mother-child interactions scored significantly higher on the ToP after intervention than before intervention. However, the gain scores of children whose mothers received the intervention were not significantly higher than those of children who received direct NDT. CONCLUSION: The results suggested that when the shared goal of parents and therapists is to enable children to express their inherent playfulness, intervention to improve parent-child interactions may be more potent than intervention directed at improving the child's developmental skills. PMID- 10686631 TI - Perceptual-motor function of school-age children with slow handwriting speed. AB - OBJECTIVES: This study investigated differences in perceptual-motor measures and sustained attention between children with slow and normal handwriting speed and the relationship between these factors. METHOD: Thirty-four slow handwriters and 35 normal speed handwriters (7 to 11 years of age) attending elementary schools in Taiwan were given three perceptual-motor tests and a vigilance task to assess sustained attention. Performances on these measures were analyzed using multivariate analysis of variance and regression analyses. RESULTS: A significant difference was found between slow and normal handwriters in upper-limb coordination, visual memory, spatial relation, form constancy, visual sequential memory, figure ground, visual-motor integration, and sustained attention. The three significant predictors of handwriting speed for the slow handwriters were age, visual sequential memory, and visual-motor integration. For the normal speed handwriters, age and upper-limb speed and dexterity were the only two significant predictors. CONCLUSIONS: Slow and normal speed handwriters responded to handwriting demands through different perceptual-motor systems. Whereas upper limb speed and dexterity seems to play an important role in normal speed handwriters, slow handwriters seem to rely more on visually directed processes, including sequence memory and visual-motor integration. PMID- 10686632 TI - Improving activities of daily living performance in an adult with ataxia. PMID- 10686633 TI - Supervisory preparedness of occupational therapists. PMID- 10686634 TI - Beyond the therapy model: building our future. PMID- 10686635 TI - Gathering current research evidence to enhance clinical reasoning. PMID- 10686636 TI - We cannot hang our hat on occupation alone. PMID- 10686637 TI - Case management practice. PMID- 10686638 TI - [Vena cava thrombosis in Behcet's disease. Analysis of a series of 10 cases]. AB - AIM: To study the clinical characteristics and the evolution of vena cava thrombosis (VCT) in Behcet's disease (BD), as well as their association with other severe symptoms. PATIENTS AND METHODS: Among 121 BD, we selected those with VCT. All patients fulfilled the diagnostic criteria of the international study group of Behcet's disease. Different clinical and paraclinical parameters were determined and compared with the remaining group of patients (not having VCT) with chi 2 test with Yates' correction. Protein C, protein S and antithrombin III and anticardiolipin antibody (aCL) levels were measured in 9 patients; anti-beta 2-glycoprotein I antibodies (a beta 2GPI) were determined in 3 patients. RESULTS: Ten patients had a vena cava thrombosis (8.2%). They were all male with an average age of 35 years (range: 30-42). We had 3 cases of superior vena cava thrombosis, 6 cases of inferior VCT, and one case of both. The average delay to diagnosis of the VCT from the date of the BD diagnosis was 4.5 years (range: 6 months-14 years), and in one case the thrombosis revealed the disease. All patients were clinically symptomatic and the installation of the symptoms were progressive and insidious in all cases. Six patients had Budd-Chiari syndrome and 4 had a phlebitis of a lower limb. Among all the clinical characteristics studied, only neurological manifestations was significantly higher in patients with VCT (p = 0.001). Protein C, protein S and antithrombin III levels were normal in all cases. One patients was positive for IgG aCL and no patient was positive for a beta 2GPI. All our patients were treated by anticoagulation therapy and high-dose prednisone combined with intravenous cyclophosphamide in 5 cases. One patient died due to liver failure. The 9 others are clinically improved (6 cases) or stable (3 cases) after an average 2.5 year course. PMID- 10686639 TI - [Motivations of requests for human immunodeficiency virus (HIV) infection screening at the Centers of Information and Anonymous and Free-of-Charge Screening. Study of 891 cases]. AB - In France, human immunodeficiency virus (HIV) testing is usually performed in centers for information and anonymous cost-free detection (Centres d'Information et de Depistage Anonyme et Gratuit, CIDAG). In this work, we studied the reasons people ask for HIV testing at the CIDAG in Lyons. Eight hundred and ninety one people were asked to give their reasons. The response ratio was 85% and the sex ratio 1:1. Seventy-five percent of the people were single. The main motivation was the desire to begin a relationship with another person without using condoms. Nonetheless, we were impressed by the high level of high-risk behaviors. To our knowledge, this is the first study about motivations of outpatients at the CIDAG. PMID- 10686640 TI - [Neuromuscular complications of long-term treatment of inflammatory diseases. 3 cases]. AB - Chloroquine and colchicine, widely used in internal medicine practice for a variety of inflammatory diseases including systemic lupus erythematosus, rheumatoid arthritis, familial Mediterranean fever, and Behcet's disease, may induce neuromuscular complications. Physicians must be familiar with this diagnosis as this iatrogenic neuromuscular pathology may simulate polymyositis, leading thus to inappropriate treatment with prednisone whereas the only effective treatment is to discontinue the drug involved whenever possible. We report three cases of toxic myopathy and/or neuropathy related to chronic chloroquine or colchicine therapy for systemic diseases, and outline the main points to be considered in this situation. PMID- 10686641 TI - Central nervous system involvement in Sjogren's syndrome: evidence from neuropsychological testing and HMPAO-SPECT. AB - OBJECTIVE: To investigate the clinical, neuropsychological and imaging manifestations of Sjogren's syndrome (SS), a chronic auto-immune disease with peripheral and central nervous system (CNS) involvement. DESIGN/METHODS: Fourteen female patients suffering from confirmed SS underwent within 2 weeks: neurological examination, immunological staging, brain MRI, brain 99m Tc-HMPAO SPECT, psychological evaluation and in-depth neuropsychological testing. RESULTS: All patients showed neuropsychological abnormalities. The cognitive symptoms were of the same type in all patients, mostly frontal lobe syndrome and memory problems. The neuropsychological involvement was not associated with other kinds of CNS involvement or MRI abnormalities, but accurately reflected HMPAO imaging results. CONCLUSIONS: The results of this study indicate that cognitive evaluation is the most sensitive clinical test to diagnose CNS involvement in patients with SS, and that CNS involvement in SS seems to be more frequent when systematically assessed by neuropsychological tests. PMID- 10686642 TI - [Familial hypercholesterolemia]. AB - Familial hypercholesterolemia is characterized by a high plasma LDL-cholesterol level. The low-density particles are the end-product of the triglyceride-rich particles, i.e. VLDL, synthetized by the liver. These triglyceride-rich particles are subsequently transformed into intermediate density lipoprotein by the lipoprotein lipase and LDL after further triglyceride hydrolysis by the hepatic lipase. The LDL particles are taken up in all cells by the mean of the LDL receptor. A large body of evidence (including experimental, clinical, epidemiological data as well as the results of large trial with lipid lowering drugs) has accumulated to establish that these particles are one of the major causative factor of atherosclerosis and its complications. Two different mechanisms may be at work in the familial hypercholesterolemia: a mutation in the LDL receptor or a single mutation in the apolipoprotein B100. Specific therapeutic intervention should be undertaken to decrease the risk to develop cardiovascular disease, mainly coronary heart disease. The therapeutic intervention includes both a diet low in saturated fatty acids and cholesterol and statins which are now the first line therapy. Fibrates are proposed to those who do not tolerate statins and LDL-apheresis is associated to statin in the rare homozygous familial hypercholesterolemia. PMID- 10686643 TI - [Human immunodeficiency virus resistance to antiretroviral drugs]. AB - Antiretroviral compounds can select viral strains presenting mutations of the HIV genome. Certain genotypic modifications are expressed by phenotypic resistance. There is no cross resistance between different classes of compounds (nucleosides, non nucleosides, antiproteases), but cross resistance is common within a given therapeutic class. HIV resistance to antiretroviral compounds is one of the principal causes of failure of antiretroviral treatments but cannot explain all escapes. The number of resistance mutations is higher in patients with high viral loads and in patients on multiple drug regimens. Currently resistance testing is limited to clinical research protocols. The usefulness of resistance testing remains to be validated. However the most eminent indications are epidemiological surveillance of primary resistance in primary infections, therapeutic adaptation after accidental exposure to HIV, and management of seropositive pregnant women. Recent retrospective studies have shown that the genotype and the phenotype after a first line treatment failure predict response to certain therapeutic combinations. In the near future, resistance testing could be useful to adapt antiviral strategies after earlier treatment failure. PMID- 10686644 TI - [What present strategies are helpful in improving transfusion safety in France?]. AB - Transfusion safety rests on measures ensuring that patients are transfused in accordance with the requirements of state-of-the-art scientific knowledge. This strict attitude is part of a quality approach which now applies to all fields of health. Transfusion is historically characterized by its ambivalence: it was the first medical discipline which integrated Quality Assurance concepts, it was also the first which proved unable to respond adequately in the face of uncontrolled risks. Today transfusion must create a system to rapidly: identify any risk, whether emergent or hypothetical; decide which action should be taken; monitor and assess corrective action; study the medico-economic impact of the whole approach. Quality assurance applies to every stage of the transfusion process, from blood donor to labile blood component recipient. This includes blood donor selection and biological control, labile blood component processing, qualification, transport and conditioning, prescription and distribution of blood components and transfused patient follow-up of. Quality controls, "safety locks", must be implemented at every stage to allow early problem detection, thus avoiding potentially dangerous attitudes and guaranteeing transfusion quality all along the process. Medical prescriptions must follow similar rules and meet Good Practice requirements defined by members of the medical and scientific community. A transfusion should not be prescribed unless it is absolutely necessary. In addition to sanitary surveillance, scientific surveillance must also be implemented to help transferring the findings of fundamental research to transfusion activities and continuously improve transfusion safety. INSERM is initiating sociological studies to identify and better understand donors' attitudes leading to risks. More sensitive tests based on nucleic acid amplification should reduce the incidence of residual viral risks. Various viral cell derivative inactivation techniques are being evaluated: the idea is to remove antigens to suppress the risk of post-transfusion alloimmunization. Numerous R&D; programs address substitution products. Transfusion safety requires all actors in the field of health being equally involved. Putting together experiences and know-how will continuously strengthen the quality approach adopted in transfusion. PMID- 10686645 TI - [Transfusion of platelet concentrates]. AB - Although a number of research have been realized in the aim of decreasing the use of platelet concentrates, these blood products remain absolutely necessary for patients with therapeutic aplasia and for some surgical patients. After the description of the main rules of platelet transfusion procedures, we will discuss some controversial issues: threshold value for platelet transfusion, platelet doses, place of curative and prophylactic strategies, refractoriness to platelet transfusion, HLA alloimmunization. Then we will focus our review on the future alternatives for platelet transfusion. PMID- 10686646 TI - Allogeneic hematopoietic stem cell transplantation: current issues and future prospects. AB - Major developments have occurred in the field of hematopoietic stem cell transplantation since the first successful transplants from HLA-identical siblings in the late 60's. The formally experimental procedure has become established therapy for a number of congenital or acquired disorders of the hematopoietic system and for chemotherapy-sensitive malignancies. Reduced transplant-related mortality has led to a widening of indications. The present review summarizes current issues and future prospects in allogeneic stem cell transplantation. These issues and prospects will include; stem cell sources, alternative donors, graft-versus-host disease, indications and long-term survival following allogeneic stem cell transplantation. PMID- 10686647 TI - [Behcet's disease: apropos of the article by N. Filali-Ansary et al]. PMID- 10686648 TI - Chronicles add value to practices and progress. PMID- 10686649 TI - The first modern operating room in America. AB - The modern OR was a vital addition to aseptic technique and essential to the development of invasive surgery, and many people do not realize that it needed to be "invented." Gustav Neuber, William S. Halsted, Charles McBurney, and others pioneered the necessary changes in practices and the environment to protect the patient from infection. This article presents the history behind the evolvement of the modern OR and discusses changes in the structures of some facilities as awareness increased about the principles of asepsis. PMID- 10686650 TI - Pharmacotherapeutics of positive inotropes. AB - Pharmacology is integrated in all areas of nursing practice. From the most basic entry level to the most advanced clinical practice, nursing curriculums are not complete without pharmacology. The word "pharmacology" often makes nurses and other health care professionals feel uneasy. Pharmacology implies complicated words and formulas that seem foreign, recalled only from one's most distant recollections of college years. For many health care providers, courses in pharmacology are taken before they care for patients, leaving them with little practical experience from which to relate. Education involves not simply learning and doing, but also applying knowledge. A basic understanding of pharmacology is needed to break down the barriers of pharmacologic communication and put the clinician at ease with terminology so often used by the medical community. PMID- 10686651 TI - Nonmetallic fixation in elective maxillofacial surgery. AB - Resorbable fixation technology offers several benefits, including easily cut and shaped plates, strong and predictable resorption qualities, and improved patient acceptance and expectations. Moreover, resorbable fixation implants can be completely reabsorbed into the body, eliminating the need for subsequent removal. This article describes the use of this innovative technology in orthognathic surgery, including preoperative and postoperative patient needs, intraoperative patient care, and potential complications. PMID- 10686652 TI - Gastrointestinal surgical patients' outcomes influenced by nutrition. AB - Nutritional support differs for patients electing to have gastrointestinal surgical procedures and patients with abdominal trauma injuries. Trauma patients can rehabilitate more quickly than patients undergoing gastrointestinal surgeries, despite the traumatic injuries that require surgical intervention. This literature review examines and evaluates the clinical practice and management of these patients and the consequences of surgical nutritional deprivation. PMID- 10686653 TI - Effective pain management in older patients. AB - Assessing and managing post-operative pain in older adult patients is complex. A baseline preoperative pain assessment provides the necessary data that assist appropriate and effective pain management. Careful monitoring and individualization of patient dosages result in effective older adult patient pain management. PMID- 10686654 TI - Recommended practices for documentation of perioperative nursing care. Association of periOperative Registered Nurses. PMID- 10686655 TI - New information on the role of beta-blockers in cardiac therapy. AB - Hypertension and ischemic heart disease are important precursors of heart failure. The prevention of progression to heart failure is a prime objective when treating patients with hypertension or ischemic heart disease. In patients with hypertension, treatment with either diuretics or beta-blockers reduces the risk of chronic heart failure. In patients with ischemic heart disease, beta-blocker therapy reduces the risk of recurrent myocardial infarction and ensuing cardiac dysfunction. The beneficial effects of beta-blocker therapy may be greater in post-infarction patients who have impaired left ventricular function than in those patients without such impairment. When considering heart failure itself, the efficacy of angiotensin-converting enzyme (ACE) inhibitors has been demonstrated in patients with mild-to-severe left ventricular dysfunction and their use is indicated for all stages of heart failure to reduce symptoms and retard further impairment of left ventricular function. Diuretics and digitalis offer relief from the symptoms of the disease, while positive inotropes are reserved for parenteral administration in end-stage heart failure, as a bridge to transplantation, or in acute exacerbations of the disease. Added to standard therapy, beta-blockade is of value in the treatment of heart failure, preventing further deterioration and improving hemodynamics, exercise tolerance, quality of life, and longterm prognosis. PMID- 10686656 TI - Antithrombotic activity of the superoxide dismutase-chondroitin sulfate complexes in a rat model of arterial injury. AB - Individual antithrombotic activities of superoxide dismutase (SOD) and sodium chondroitin sulfate (CHS) as well as the activities of covalent and noncovalent complexes of SOD with CHS were compared in a rat model of arterial thrombosis induced by ferrous chloride. Covalent conjugate of SOD with CHS exerted the most potent antithrombotic effect, which was associated with adsorption of the conjugate on the glycocalyx of the vascular wall cells and stability of the covalent bond between CHS and SOD subunits. Theoretical and practical directions in the investigation of SOD and CHS preparations are outlined. PMID- 10686657 TI - Effect of low-dose aspirin on the markers of oxidative stress. AB - The present study estimates effects of low-dose enteric coated aspirin (ECA) on oxidative stress (OS) markers in a group of middle-aged men (mean age 51.2 +/- 6.9 years) free of pre-existing ischemic heart disease. METHODS: Serum products of lipid peroxidation, and measures of antioxidative status were detected in 25 healthy men in baseline and after two-week treatment period. RESULTS: In respect to serum products of lipid peroxidation and markers of antioxidant status, no statistically significant differences between the pre- and after-treatment data were observed for any measures, with the exception of values of serum antioxidative capacity (39.0 +/- 2.5 and 42 +/- 4.6, respectively). CONCLUSIONS: Administration of ECA does not initiate the OS in blood and improves the general antioxidative potency of blood. This may imply towards certain antiatherogenic influence of low-dose ECA, exhibited even with a short-term treatment period. Regarding OS markers, a variety of individual responses observed in the selected subgroups should be investigated and possibly taken into account while treatment with ECA is initiated for primary prevention of cerebrovascular events. PMID- 10686658 TI - Pharmacological modification of the dispersion of repolarization in the heart: importance of the M cells. AB - Several in vitro and in vivo investigations have provided data supporting the existence of M cells in the deep subepicardial layers of the ventricles in a number of species. Characterized by unique electrophysiological and pharmacological features, this population of cells is regarded to have a significant role in creating dispersion of repolarization in the ventricular wall and thus contribute importantly to arrhythmogenesis, in particular to intramural reentry and triggered activity. Focusing on M cells, the authors summarize recent findings and concepts concerning the pharmacological heterogeneity of different cell and tissue types found within the ventricles and explore how these differences may contribute to electrocardiographic manifestations. On the basis of literary data and of their own results they conclude that studying the electrical and pharmacological inhomogeneity within the ventricular wall may provide a better understanding of the pathophysiological processes that give rise to cardiac rhythm disturbances and the mechanisms by which antiarrhythmic agents act to suppress and in some cases aggravate arrhythmias. PMID- 10686659 TI - Evaluation of atrial refractoriness and atrial fibrillation inducibility immediately after internal cardioversion in patients with chronic persistent atrial fibrillation. AB - OBJECTIVE: To prospectively evaluate right atrial refractoriness and sustained atrial fibrillation (AF) inducibility at programmed electrical stimulation in two groups of patient: a series of patients with chronic persistent AF, studied immediately after successful low energy internal atrial cardioversion, and a group of control patients without history of supraventricular arrhythmias. PATIENTS: Nineteen patients with chronic persistent AF (mean AF duration 11 +/- 10 months, range 2-61 months) submitted to successful internal low energy atrial cardioversion in fully conscious state and 11 control patients without history of supraventricular arrhythmias. METHODS: An electrophysiological evaluation was performed to measure atrial refractoriness and AF inducibility, by delivering single atrial extrastimuli in high right atrium, at decremental coupling, during spontaneous sinus rhythm and after 8 beats at 600, 500, 400 and 330 ms cycle length. If sustained AF was induced the protocol was terminated. RESULTS: During programmed atrial stimulation sustained AF was induced in 8 out of 19 (42%) of the AF patients but in none of the control group. Atrial effective refractory period was significantly shorter in AF patients compared to controls both at basic cycle length, at 600 ms, 500 ms and 400 ms cycle length, meanwhile no statistically significant differences were found at 330 ms cycle length. An altered relationship between atrial effective refractory period and cycle length was found in AF patients compared to controls: the slope of linear correlation slope was significantly lower in AF group than in controls (0.04 +/- 0.07 vs 0.17 +/- 0.10, p < 0.002). CONCLUSIONS: Marked abnormalities of atrial refractoriness and of its heart rate relationship are observed after internal cardioversion of chronic persistent AF in humans and these abnormalities are associated with an high vulnerability to AF. These observations may explain the high risk of AF recurrences in the early phases following successful cardioversion. In this scenario antiarrhythmic drug therapy seems to be mandatory for reducing arrhythmia relapses. PMID- 10686660 TI - Amlodipine and physiological responses to brisk exercise in healthy subjects. AB - Recent studies have questioned the safety of calcium antagonists in general, and short-acting dihydropyridine derivatives in particular. Reasons include excessive catecholamine stimulation after stress. We therefore wanted to assess whether amlodipine, a second generation dihydropyridine with a prolonged plasma half life, would show a more favourable haemodynamic and biochemical profile after strenuous exercise. For this purpose, we studied 9 healthy volunteers in a double blind, randomized, placebo-controlled trial. After 10 days of amlodipine, 5 mg orally daily or placebo therapy, volunteers performed a treadmill effort test; the sequence was repeated after a 2-week washout period. Amlodipine caused a significant increase in mean resting heart rate (HR) (from 65 +/- 3 to 70 +/- 3 beats/min, p < 0.05), without changing systolic or diastolic blood pressure (SBP, DBP). Post-exercise haemodynamic responses were similar while on amlodipine or placebo therapy. Amlodipine did not alter the normal profile of resting or exercise-induced metabolic [plasma glucose, serum K+, serum free fatty acid (FFA)] and hormonal [plasma cortisol, growth hormone, prolactin, insulin, epinephrine (EPI) and norepinephrine (NE)] responses--although plasma EPI concentrations dropped significantly lower (p < 0.05) at 5 min and 15 min post exercise while on the calcium antagonist. We conclude that amlodipine has a largely neutral effect on the physiological profile after brisk exercise in healthy young subjects and that this may prove to be a useful property for a vasodilator drug. PMID- 10686661 TI - Cilnidipine improves insulin sensitivity in the Otsuka Long-Evans Tokushima fatty rat, a model of spontaneous NIDDM. AB - Among the antihypertensive drugs, fast-acting Ca2+ antagonists have been reported to worsen insulin sensitivity. This effect may be attributable to reflex increases in sympathetic activity. On the other hand, however, it has been reported that long-acting, dihydropiridine Ca2+ antagonists improve insulin resistance. The purpose of this study was to investigate whether cilnidipine, another long-acting dihydropidine Ca2+ antagonist, improves insulin sensitivity in Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model of spontaneous NIDDM. 25 weeks OLETF rats were divided into the following groups; normal-diet group, cilnidipine-supplemented group (cilnidipine 3 mg/kg/day) and angiotensin II receptor antagonist CS-866-supplemented group (CS-866 1 mg/kg/day). As a non diabetic control, we used Long-Evans-Tokushima-Otsuka rats (non-diabetic rats). Glucose infusion rate (GIR), an index of insulin resistance, as measured by the hyperinsulinemic euglycemic clamp technique was significantly decreased in OLETF rats. Cilnidipine-treatment partially but significantly improved insulin sensitivity in addition to systolic blood pressure in OLETF rats at 30 weeks of age, although it did not decrease accumulation of abdominal fat or serum levels of glucose or insulin. CS-866, an angiotensin II receptor antagonist, which lowers blood pressure through a different mechanism, did not improve insulin resistant states in OLETF rats. These results suggest that cilnidipine has a beneficial effect on insulin-resistance together with the antihypertensive effect. PMID- 10686662 TI - Cardioprotective efficacy of verapamil and mibefradil in young UM-X7.1 cardiomyopathic hamsters. AB - Since calcium overload and increased in T-type calcium channel activity have been observed in the cardiomyopathic (CM) hamster, we hypothesized that mibefradil (Ro 40-5967), a new T- and L-type calcium channel blocker, may exert significant cardioprotection in the early phase of the disease. Young (30-day-old) CM hamsters of the UM-X7.1 subline were treated with mibefradil or verapamil for 4 to 6 weeks. Mibefradil doses were in the range of 0.5 to 8 mg/kg/day while verapamil was given at a dose of 5-10 mg/kg/day, both drugs being injected twice daily (s.c. and i.p. alternatively). At the end of the treatment period, myocardial and skeletal muscle (tongue) were harvested and processed for assessment of necrotic changes and calcification. In hearts from control CM hamsters, numerous necrotic and calcified foci were observed. These myocardial necrosis markers were not attenuated by mibefradil in the dose range studied whereas verapamil significantly reduced their severity. The dystrophic process in skeletal muscle (tongue) was not inhibited by mibefradil or verapamil. These results suggest that mechanisms other than inhibition of T- and L-type calcium channels are related to the cardioprotection observed in the presence of verapamil. A specific action on the sarcoplasmic reticulum (ryanodine-sensitive calcium channel) or the mitochondria may explain the efficacy of phenylalkylamines (verapamil) in this condition. PMID- 10686663 TI - How long can an escalation of dose override tolerance to the hypotensive efficacy of nitroglycerin infusion in coronary care patients. AB - Tolerance to nitroglycerin infusion (NG) can be overridden by dose escalation. The aim of this study was to define for how long it can be done for hypotensive efficacy of NG, in a coronary care setting. A prospective trial with an intra individual therapeutic comparison was performed in 60 patients with acute myocardial infarction or unstable angina. Initial efficacy of NG was confirmed by a 10% blood pressure decrease (measured by cuff). Seventy-two-hour NG infusion was interrupted, for 30 minutes, every 12 hours. If blood pressure increased by 10% after infusion interruption, the infusion was continued at the previous rate. If blood pressure did not increase (detected tolerance--weakened efficacy of NG), the dose was increased until pressure decreased by 10% and the infusion was continued at the new dose. Failure to achieve hypotensive response, despite a 5 fold dose increase, indicated onset of resistance--completely lost hypotensive efficacy of NG. The majority of patients (49 out of 55) who developed tolerance, developed it during the first 36 hours, while the majority of those who developed resistance (33 out of 40), developed it within 60 hours of the infusion. Tolerance was overridden by dose escalation in 41 out of 55 patients, which was repeated in 31 patients. Complete restoration of NG action was possible over 24 hours in half the patients, and over 48 hours in one third of the patients. Three out of 34 patients who developed tolerance before the 13th hour did not develop resistance during the following 60 hours of dose up-titration. The conclusion is that tolerance to NG can be overridden by dose escalation in the majority of patients for a significant period of time, which is useful in clinical practice. PMID- 10686664 TI - Prolonged oral L-carnitine substitution increases bicycle ergometer performance in patients with severe, ischemically induced cardiac insufficiency. AB - Acute and chronic L-carnitine application exerts protective effects in a number of cardiac diseases. These favourable effects are attributed to improvements of the energy metabolism and have been found both in animal experiments and in man. In order to investigate the effect of long-time oral L-carnitine substitution on physical performance, 41 patients suffering from class NYHA II or III cardiac insufficiency were recruited for a clinical study. Following the double-blind, randomized, placebo-controlled design of the study, 20 patients were given 3 x 1 g L-carnitine daily for 120 days whereas the control group (21 patients) received placebo. Bicycle ergometer tests were used to determine maximum performance, systolic and diastolic blood pressure, heart rate, and ST changes. Four series of tests were carried out: on day 0 (before the first substrate application), on the 60th and the 120th day (during L-carnitine or placebo application), and on the 180th day (60 days after the end of substitution). A significant improvement in performance (significantly higher maximum performance during bicycle ergometry) could be found within the carnitine group on the 60th and 120th day of L carnitine application; and haemodynamical parameters showed a tendency to improve, too. These effects, which were attributed to L-carnitine, could be detected even 60 days after the end of substitution. No corresponding changes were found in the placebo group. The findings presented in this paper support suggestions of other authors that L-carnitine in combination with the usual medication (digitalis, beta-blockers, calcium antagonists, nitrates) improves performance and effort tolerance in patients with cardiac insufficiency. Moreover, the findings suggest a favourable long-term effect, which lasts beyond the actual L-carnitine application, on the performance of patients with advanced cardiac insufficiency. PMID- 10686665 TI - Efficacy of rilmenidine versus captopril on microalbuminuria: a pilot study in hypertensive type 2 diabetics. PMID- 10686666 TI - The effect of nebivolol on left ventricular hypertrophy in hypertension. PMID- 10686667 TI - Experimental susceptibility of turbot Scophthalmus maximus to viral haemorrhagic septicaemia virus isolated from cultivated turbot. AB - Juvenile pathogen-free turbot were infected with a viral haemorrhagic septicaemia virus (VHSV) isolate recovered from turbot cultivated on the island of Gigha, West Scotland. Mortality of 100% was recorded in fish infected via the intra peritoneal (i.p.) route. Horizontal transmission of VHSV in sea water was demonstrated by cohabitation of naive fish with i.p. infected fish at a ratio of 1:1. The total cumulative average mortality in cohabiting fish was 60% by 60 d post-infection. Turbot infected via an immersion route exhibited a cumulative average mortality of 71% by the end of the experiment. VHSV identified by enzyme linked immunosorbent assay (ELISA) was recovered from both organ (kidney and spleen) and brain samples of individual fish that died following infection by all experimental routes. These findings pose significant implications regarding the persistence of VHSV and its role in limiting natural populations of marine fish species. In addition, the establishment of infection models for the transmission of VHSV in sea water is of fundamental importance to the development of anti-VHSV vaccines in important commercial species such as turbot. PMID- 10686668 TI - Pathogenicity of nodavirus strains from striped jack Pseudocaranx dentex and Atlantic halibut Hippoglossus hippoglossus, studied by waterborne challenge of yolk-sac larvae of both teleost species. AB - The present study shows that differences in pathogenicity exist among fish nodavirus strains. In challenge trials, a Japanese strain (SJ93Nag) was highly virulent to larvae of the striped jack Pseudocaranx dentex but replication was not detected in larvae of Atlantic halibut Hippoglossus hippoglossus at 6 degrees C. Conversely, a Norwegian nodavirus strain (AH95NorA) that was highly virulent to the Atlantic halibut larvae did not replicate in striped jack larvae at 20 degrees C. Occurrence of the disease viral encephalopathy and retinopathy (VER) and cumulative mortality were significantly different in the 2 species when challenged with the 2 nodavirus strains. The presence of nodavirus in nervous tissue was monitored by immunohistochemical methods. Our results support the view that the genetic diversity among nodavirus strains reflects the existence of different viral phenotypes which may be adapted to infect different host species and/or for replicating at different temperatures. Fish nodaviruses represent surveyable pathogens well suited for studying the relation between viral genotypic and phenotypic properties such as host specificity, temperature optima, neuroinvasiveness and neurovirulence. PMID- 10686669 TI - Naked DNA vaccination of Atlantic salmon Salmo salar against IHNV. AB - A naked plasmid DNA encoding the glycoprotein (pCMV4-G) of a 1976 isolate of infectious hematopoietic necrosis virus (IHNV) obtained from steelhead Oncorhynchus mykiss was used to vaccinate Atlantic salmon Salmo salar against IHNV. Eight weeks post-vaccination the fish were challenged with a strain of IHNV originally isolated from farmed Atlantic salmon undergoing an epizootic. Fish injected with the glycoprotein-encoding plasmid were significantly (p < 0.05) protected against IHNV by both immersion and cohabitation challenge. Survivors of the first challenges were pooled and re-challenged by immersion 12 wk after the initial challenge. Significant (p < 0.05) protection was observed in all of the previously challenged groups including those receiving the complete vaccine. Fish injected with the glycoprotein-encoding plasmid produced low levels of virus neutralizing antibodies prior to the first challenge. Neutralizing antibodies increased in all groups after exposure to the IHNV. Passive transfer of pooled sera from pCMV4-G vaccinates and IHN survivors provided relative survivals of 40 to 100% compared to fish injected with sera collected from fish immunized with control vaccines or left unhandled. In this study, DNA vaccination effectively protected Atlantic salmon smolts against challenges with IHNV. PMID- 10686670 TI - Comparative severity of experimentally induced mycobacteriosis in striped bass Morone saxatilis and hybrid tilapia Oreochromis spp. AB - Twenty striped bass Morone saxatilis and 20 hybrid tilapia Oreochromis niloticus x O. mossambicus x O. aureus each received a single intramuscular injection of 1.6 x 10(6) colony forming units per gram body weight of Mycobacterium marinum. Striped bass manifested significantly greater clinical and microscopic disease compared to tilapia. Whereas all the striped bass had died or were clinically ill by Day 8 post-infection, there was no apparent disruption of normal behaviour, physical appearance, or growth in any of the sacrificed or surviving tilapia. Histologically, granulomas in striped bass were generally larger and less discrete, with a higher proportion of heavily vacuolated macrophages, and large cores of necrotic cells. Visceral granulomas in tilapia were smaller, with a higher proportion of epithelioid macrophages, more pigment-containing cells, more peripheral lymphocytes, and virtually no central necrosis. Visceral granulomas were 18-fold more numerous in striped bass than in tilapia. Based upon histomorphometric data, mean proportions of acid-fast bacteria within pronephros granulomas were 4-fold greater in striped bass than tilapia, and striped bass granulomas averaged more than twice as large as tilapia granulomas. In the anterior kidney of striped bass, a positive correlation existed between mean mycobacterial proportions and mean necrosis scores. In tilapia, mean mycobacterial proportions correlated negatively with mean granuloma numbers, whereas there was no correlation between these parameters in striped bass. Results suggest that intrinsic functional differences in the immunologic systems of striped bass and hybrid tilapia may contribute to inter-species variation in mycobacteriosis susceptibility. PMID- 10686671 TI - Infectious necrotizing enteritis and mortality caused by Vibrio carchariae in summer flounder Paralichthys dentatus during intensive culture. AB - An epizootic causing mortality among cultured summer flounder Paralichthys dentatus occurred in summer of 1998 at a land-based facility on Narragansett Bay, Rhode Island, USA. The disease, flounder infectious necrotizing enteritis (FINE), was characterized by reddening around the anal area, distended abdomens filled with opaque serosanguineous fluid, enteritis and necrosis of the posterior intestine. In extreme cases of the disease, the posterior intestine was detached from the anus and was observed coming out the vent. The intestine of individuals that recovered from the disease ended in a blind-sac; the abdomens of these fish were distended, due to food and water inside the intestinal blind-sac. A bacterium was isolated from ascites fluid and kidney of moribund flounder and identified as the causative agent in challenge experiments. The pathogen was identified as Vibrio carchariae by morphological and biochemical characteristics and sequence of the 16S rRNA. The LD50 estimate was 5 x 10(5) colony-forming units injected intraperitoneally into 100 to 200 g summer flounder. PMID- 10686672 TI - Experimental detection of the actinospores of Myxobolus pseudodispar (Myxosporea: Myxobolidae) in oligochaete alternate hosts. AB - The development of Myxobolus pseudodispar Gorbunova, 1936, an intracellular myxosporean muscle parasite of the roach Rutilus rutilus L., was studied in experimentally infected oligochaetes. In one experiment, uninfected Tubifex tubifex Muller and Limnodrilus hoffmeisteri (Claparede) were exposed to mature spores of M. pseudodispar. Triactinomyxon spores developed both in T. tubifex and L. hoffmeisteri specimens. Triactinospores were first released from the oligochaetes 76 d after initial exposure. At that time, pansporocysts containing 8 triactinospores were located in the gut epithelium of experimentally infected oligochaetes, but free actinosporean stages were also found in their gut lumen. Each triactinospore had 3 pyriform polar capsules and an elongated cylindrical sporoplasm with 8 secondary cells. The spore body joined the 3 caudal projections with a relatively long style. One of the 3 caudal projections was shorter than the other two. The total length of the triactinospore was on average 206.5 microns. PMID- 10686673 TI - Henneguya ghaffari sp. n. (Myxozoa: Myxosporea), infecting the Nile perch Lates niloticus (Teleostei: Centropomidae). AB - Light microscopical description is presented for a new myxozoan species, Henneguya ghaffari, which infects the Nile perch Lates niloticus (Linnaeus, 1758) in Lake Wadi El-Raiyan in Egypt. The spore is characterized by a triangular thickening at the base of the caudal processes. The relatively long caudal processes run adherent to each other for two-thirds of their length, then bifurcate to very fine processes. Prevalence of infection was 34.6% and peaked during winter and early spring. The infection was concentrated along the intestinal tract, and in severe cases gills and gill rakers were also infected. Histology revealed that, in contrast to findings of previously published works on related species, intralamellar plasmodia did not develop inside the blood capillaries of the gills. Intestinal plasmodia were very pathogenic due to their large number and size. These plasmodia caused atrophy of the muscularis layer, and replaced and distended the submucosal and mucosal layers. The validity of some Henneguya species in Africa is discussed. PMID- 10686674 TI - Absence of vertical transmission of infectious salmon anemia virus (ISAV) from individually infected Atlantic salmon Salmo salar. AB - Atlantic salmon Salmo salar L. eggs were collected from grilse that were individually identified as ISAV-positive based on the detection of pathogen in ovarian fluid by RT-PCR. The eggs were fertilised, disinfected and reared under quarantine conditions. To address the possibility of vertical transmission, fertilised eggs, alevins and parr were screened for the virus by SHK-1 cell culture and RT-PCR. In addition, ISAV-negative parr were injected with homogenates of potentially infected eyed eggs. ISAV was not detected in eyed eggs, alevins or parr. No mortalities occurred among fish injected with the egg homogenates. These observations suggest the absence of a vertical transmission route for ISAV infection. PMID- 10686676 TI - [Multiple sclerosis--a disease that never sleeps. Satellite Symposium "Multiple sclerosis must be treated: early start of interferon therapy" within the framework of the 72nd Annual Meeting of the German Society for Neurology. Magdeburg, 30 September 1999]. PMID- 10686675 TI - First record of Emetha audouini, a cymothoid isopod parasite, from cultured sea bass Dicentrarchus labrax in Greece. AB - For the first time, Emetha audouini (Milne Edwards, 1840), a cymothoid isopod, is reported parasitising cage cultured sea bass Dicentrarchus labrax L., 1758 in Greece. The specimens observed are larvae (Pulli II). They were found in great numbers in the buccal and branchial cavity of young (3.5 m.o. old) sea bass, in an intensive cage farm facility. This parasite is certainly transferred to sea bass from wild populations of Sparidae or Centracanthidae. Serious lesions were visible and typical of a crustacean infection, with extensive and deep skin damage in the head area. The cumulative mortality, over a 2 wk period, was 10.75%. The parasitic problem was successfully dealt with by optimization of management practices rather than use of costly and dangerous chemotherapeutants. PMID- 10686677 TI - [Challenges in epilepsy therapy: start low, go slow. 72nd Congress of the German Society for Neurology. Topiramate--a valuable new tool for the therapy of focal epilepsy? 1 October 1999, Magdeburg]. PMID- 10686678 TI - [Alzheimer dementia and depression--a therapeutic challenge. In comorbidity they burden the Alzheimer patient as well as his caretakers]. PMID- 10686679 TI - [The cardiologist and first aid: the situation in Italy]. PMID- 10686680 TI - The experience of chest pain units. PMID- 10686681 TI - [Is it possible to reformulate the strategic approach to chest pain in Italy today?]. PMID- 10686682 TI - [Organizational strategy for cardiac emergencies during the 2000 Jubilee]. PMID- 10686684 TI - Prehospital fibrinolysis. PMID- 10686683 TI - [The avoidable delay]. PMID- 10686685 TI - [Hospital network for primary PTCA: the Italian situation]. PMID- 10686686 TI - Prehospital triage and reperfusion therapy for acute myocardial infarction. PMID- 10686687 TI - [Prognostic stratification]. PMID- 10686688 TI - [Management of acute coronary syndrome. New pharmacologic approaches]. PMID- 10686689 TI - [Role of revascularization in acute coronary syndrome]. PMID- 10686690 TI - The management of cardiogenic shock. PMID- 10686691 TI - [The first two links of the "survival chain": the 118 system and early emergency care]. PMID- 10686692 TI - [The third and fourth link of the chain: defibrillation and advanced support]. PMID- 10686693 TI - [Is it possible to identify patients at risk of sudden cardiac death?]. PMID- 10686694 TI - [Intervention tools in patients at risk of fatal arrhythmia: medical treatment or ICD. Indicators and results]. PMID- 10686695 TI - [Life Project: the first European project of early defibrillation with first responders]. PMID- 10686696 TI - [Dimensions of the educational problem]. PMID- 10686697 TI - [Role of national health institutions and of professional societies]. PMID- 10686698 TI - [Medical-scientific information]. PMID- 10686699 TI - [Continuing education: legislative basis for a possible development]. PMID- 10686700 TI - [Cardiovascular treatment: point of view of the cardiologist and the controlling body]. PMID- 10686701 TI - [Thrombolytic therapy in the year 2000]. PMID- 10686702 TI - [Fight against sudden death in Italy: what can be done and what should be done]. PMID- 10686703 TI - National Committee for the Coordination of Antibiotic Policy in Belgium: the next step in the fight against antimicrobial resistance. PMID- 10686704 TI - Prevention of pneumococcal disease: an update on the Belgian Consensus Report. AB - An ad hoc working party on pneumococcal vaccine with representatives of the Belgian Society for Infectiology and Clinical Microbiology, the Belgian Society of Pulmonology and Scientific Societies of General Practitioners reviewed new data on the epidemiology of S. pneumoniae infections in Belgium, on the efficacy and the cost-effectiveness of the 23-valent capsular polysaccharide vaccine. We discuss recent data on vaccination with a new conjugate pneumococcal vaccine, shown to be highly effective in children. The Working Group of the Belgian Scientific Societies endorses the recommendations issued by the Hoge Gezondheidsraad in 1993 and described in a consensus report in 1996. PMID- 10686705 TI - Glycaemic and blood pressure controls achieved in a cohort of 318 patients with type 2 diabetes. AB - The aim of the study was to evaluate the current antihyperglycaemic and antihypertensive treatment schemes as well as the quality of metabolic control and blood pressure in a population with type 2 diabetes, in view of the United Kingdom Prospective Diabetes Study (UKPDS) data. 318 patients were included. 44% were treated with metformin and/or sulfonylurea. 44% received insulin in monotherapy or combined with oral drugs. HbA1c was 8.0 (7.9-9.4)% (median; percentiles 25-75). Chronic neurological and vascular (micro- and macroangiopathy) complications were present in 21-43% of patients and were related to glycaemic control. (Un)treated hypertension was found in 59% of patients. Main treatments were ACE-inhibitors (40%), calcium channel antagonists and diuretics (20%) and/or beta-blockers (18%). Systolic and diastolic blood pressure were 147 +/- 22 and 86 +/- 12 mm/Hg (mean +/- 1 SD). In conclusion, overall glycaemic control of a type 2 diabetic population remains slightly unsatisfactory in view of the UKPDS recommendations. In contrast, blood pressure control was adequate. PMID- 10686706 TI - [Endocrine complications of genetic hemochromatosis]. AB - The authors report the prevalence and severity of endocrine complications in a cohort of 115 patients suffering form genetic hemochromatosis and followed since two decades. Already 40% of them had developed diabetes at the time of diagnosis of hemochromatosis, which was made at the age of 50 +/- 12 years (m +/- SD). Hypogonadism was evidenced in 42% of the patients. In most of them, it was considered as secondary to pituitary lesions as assessed by GnRH tests. No other endocrine complications, in particular thyroid disease, were evidenced during follow up. On the other hand, it was also of interest to note that liver cirrhosis was observed in 52% of the patients at the time of hemochromatosis diagnosis. A relationship between cirrhosis, diabetes and hypogonadism was also assessed in these patients. We conclude to the still high prevalence of endocrine complications in genetic hemochromatosis. PMID- 10686707 TI - Persisting misconceptions of Belgian physicians and nurses about cancer pain treatment. AB - In Belgium palliative hospices, palliative support teams in hospitals and palliative home care are well-developed. The author gave a lecture about pain treatment in palliative care and inquired after the knowledge and attitudes of 28 nurses and 45 physicians. A questionnaire containing questions about morphine and cancer pain treatment was completed by the attendees before and after the session. In the initial questionnaire the care-providers' attitudes towards palliative care and symptom control were included also. The Wilcoxon test revealed a significant difference in knowledge between the physicians and nurses before the session (p = 0.007). Afterwards knowledge had improved in both groups (p = 0.007) but a difference still remained (p = 0.007). This study reveals that continued education is mandatory. An oral presentation seems not ideal; interactive training with practical exercises might be more appropriate. PMID- 10686708 TI - Diagnosis and treatment of an unusual cause of metabolic acidosis: ethylene glycol poisoning. AB - Ethylene glycol intoxication is a rare but dangerous type of poisoning. It causes a severe acidosis with high anion and osmolal gaps. Clinical manifestations of the ethylene glycol intoxication can be divided in three phases: a neurologic stage, with hallucinations, stupor and coma; the second stage is cardiovascular with cardiac failure. Renal failure characterizes the third stage, due to acute tubular necrosis. After aggressive gastric emptying, the main treatment is ethanol or 4-methypyrazole, which can be given either orally or intravenous, with supportive measures for all symptoms or diseased organ. PMID- 10686709 TI - Acromegaly and Waldenstrom's macroglobulinaemia. PMID- 10686710 TI - [Thrombocytopenia and lupus]. PMID- 10686711 TI - [Idiopathic thrombocytopenic purpura as first manifestation of systemic lupus erythematosus lupus]. AB - OBJECTIVES: We studied SLE whose initial symptoms were related to idiopathic thrombocytopenic purpura (ITP) in order to determine if they share clinical and immunologic manifestations. METHODS: We reviewed the clinical backgrounds of 150 SLE (according to the ACR criteria) periodically followed from 1980 until 1998. We found 12 patients with both these conditions. RESULTS: All patients were female with a mean age of 32 at the time of ITP diagnosis and 36 at the time of SLE diagnosis. The most usual clinical manifestations were: arthritis (92%), cutaneous (58%) and hematologic involvement with lymphopenia (58%) and thrombocytopenia again (33%) after the initial ITP episode, always together with autoimmune hemolytic anemia (Evans syndrome). None of these patients presented with neurologic involvement and only one presented with renal involvement. 50% were positive for anti-DNA antibodies, 50% were Ro(+) and 16% were RNP (+). 66% were positive for antiphospholipid antibodies and 33% for lupus anticoagulant. Thrombocytopenia was controlled just with steroids in only 16% of the patients. Splenectomy controlled thrombocytopenia with complete remission achieved in 80% (4 from 5) of the patients and 20% (1 from 5) were refractory to this therapy after a medium follow-up time of 6.5 years. CONCLUSIONS: 1) SLE whose initial symptoms are related to ITP were characterized by joints, cutaneous and hematologic involvement without renal and neurologic manifestations. 2) Splenectomy was able to control refractory thrombocytopenia in the majority of these patients. PMID- 10686712 TI - [Secondary amyloidosis in rheumatoid arthritis. A clinical study of 29 patients]. AB - BACKGROUND: To study the clinical features, outcome and response to therapy in 29 cases of rheumatoid arthritis (RA) with secondary amyloidosis (AA). PATIENTS AND METHOD: Twenty-nine patients with RA and AA who were diagnosed during 11 years. RESULTS: The mean age and the mean duration of RA were 63 +/- 12 years and 15 +/- 7 years respectively. The most common initial clinical feature was renal involvement (83%). Nineteen patients were treated with methotrexate. The mean survival time was 42 +/- 8 months. Eleven patients (38%) have deceased. CONCLUSION: Our data confirms that AA in RA is a serious complication with a bleak prognosis. A normal renal function is a good prognosis indicator. PMID- 10686714 TI - [Pernicious anemia and other megaloblastic anemias]. AB - OBJECTIVE: To describe the clinical and biological characteristics of a series of patients with megaloblastic anemia (MA) and to identify potential differences between patients with pernicious anemia (PA(+)) and patients with other MA (PA( )). METHOD: Retrospective study of 50 patients with MA diagnosed in our service between 1993 and 1998. RESULTS: MA was diagnosed in 50 patients. The median age in the moment of diagnosis was 70.7 years. The causes of MA were: cobalamin deficiency (CD) in 40 cases (80%), folate deficiency (FD) in 7 cases (14%) and both deficiencies in 3 cases (6%). PA was diagnosed in 19 patients (38%). All cases showed hyper-segmented neutrophils and 41 cases (81%) macroovalocytosis. Hemoglobin level < 8 g/dl was present in 22 patients (44%). The median serum LDH level was 2.059 +/- 1.739 U/l. There was a lower frequency of female sex and a higher RDW in the group PA(+). There were no significant differences between both groups in the rest of studied features, except for the presence of antiparietal cell antibodies and anti-intrinsic factor antibodies in the group PA(+). CONCLUSIONS: CD was the most frequent cause of MA in our series. PA was the most frequent cause of CD. Most cases of MA corresponded to a severe macrocytic anemia with hyper-segmented neutrophils, macroovalocytosis and very high serum LDH level. We did not identify any clinical or biological characteristic, except for the presence of antiparietal cell antibodies and anti-intrinsic factor antibodies and a higher RDW in the group PA(+), to permit distinguish the groups PA(+) and PA(-). PMID- 10686713 TI - [Efficacy of hygienic and dietary therapy in coronary patients with isolated hypoalphalipoproteinemia]. AB - OBJECTIVE: Hypoalphalipoproteinemia (HA) is a relatively frequent disorder found in patients with coronary artery disease (CAD). It is associated to a greater risk of suffering recurrent coronary episodes and of mortality caused by this disease. METHODS: We selected 60 patients with previous CAD and isolated HA (HDLc concentration < 0.9 mmol/L, and desirable lipidic profile) that were consecutively seen in a specialized lipid clinic. Subjects were randomly included in the two groups of cases (group of intervention) and controls. Cases were treated with non-pharmacological measures which included changes in lifestyle and dietary habits. Control subjects were referred to their general practitioners in order to receive conventional medical care. RESULTS: It was demonstrated a significant increase in the HDLc concentration in both groups, being greater the improvement in the group of intervention, but the differences in the increase in the HDLc between both groups were not significant. Fibrinogen was lower in the patients of the group of intervention, especially in those patients that gave up smoking. CONCLUSION: Changes in lifestyle and dietary habits are useful to correct the low HDLc plasma levels and to reduce fibrinogen levels in those patients with CAD and HA. PMID- 10686715 TI - [Gronblad-Strandberg syndrome. Report of four cases in one family]. AB - The Gronblad-Strandberg syndrome is a rare congenital hereditary dysplasic disorder of the connective tissue, characterized by a progressive abnormal mineralization and dystrophic calcification of elastic tissue and collagen. This process affects tissues rich in elastic fibers and multiple systems of the organism, the cutaneous, ocular and vascular being the most common. These findings progress through the life of the affected person. We present 4 cases in the same family, with heterogeneous clinic pattern and evolution. PMID- 10686716 TI - [Endocarditis caused by Actinobacillus actinomycetemcomitans]. AB - The HACEK group endocarditis are infrequent in general population. They usually develop subacute endocarditis with large vegetations, peripheral emboli, heart failure, requiring frequent valvular replacement. We report a clinical case of endocarditis caused by Actinobacillus actinomycetemcomitans (AA), without the typical findings of the HACEK group endocarditis and with a successful evolution with medical treatment. PMID- 10686717 TI - [Cellulitis caused by Aeromonas hydrophila]. AB - We report the case of a cirrhotic patient with leukocytoclastic vasculitis who developed a rapid and progressive cellulitis with hemorrhagic bulla and sepsis due to Aeromonas hydrophila, the portal of entry was the surgical leech of a cutaneous biopsy. PMID- 10686718 TI - [Pasteurella spp: a new microorganism to the cause of acute suppurative thyroiditis]. AB - Acute suppurative thyroiditis is an uncommon disease due to local resistance of the gland to infection. Preexisting gland pathology and local anatomic abnormalities are predisposing factors. We present the first case described in the medical literature caused by Pasteurella spp after upper respiratory infection with insidious manifestations resembling subacute thyroiditis. The course was benign after surgical drainage. PMID- 10686719 TI - [Gronblad-Strandberg syndrome]. AB - The Gronblad-Strandberg syndrome, also known as pseudoxanthoma elasticum, is a rare congenital dysplasic disorder of the connective tissue, characterized by a progressive abnormal mineralization and dystrophic calcification of elastic tissues and collagen (1). The mode of inheritance is uncertain, but autosomal recessive inheritance can be assumed. This process affects tissues rich in elastic fibres and multiple systems of the organism, being the most common the cutaneous (pseudoxanthoma elasticum), ocular (angioid streaks), vascular (occlusive vascular disease) and gastrointestinal manifestations. These findings progress through the life of the affected person, with variable pattern clinic and evolution. PMID- 10686720 TI - [Prions: definition and diseases]. AB - In this article we review the concept and terminology of prions, their replication and some current hypothesis on the nature of these infectious agents causing neurodegenerative diseases. This revision also summarizes the etiopathogenic, epidemiological, clinical and neuropathological features of the prion diseases or human transmissible spongiform encephalopathies, and some methods for their early diagnosis. Finally, we discuss the possible link between the bovine spongiform encephalopathy and the new cases of Creutzfeldt-Jakob disease identified in the United Kingdom. PMID- 10686721 TI - [Itraconazole in the treatment of invasive aspergillosis. A case report]. PMID- 10686722 TI - [A new case of association of multiple myeloma and megaloblastic anemia]. PMID- 10686723 TI - [Multiple myeloma and megaloblastic anemia]. PMID- 10686724 TI - [Herpetic tracheobronchitis in a patient with the acquired immunodeficiency syndrome]. PMID- 10686725 TI - [Mucinous cystadenocarcinoma of the recto-appendiceal fistula]. PMID- 10686726 TI - [Primary and secondary prophylaxis of venous thrombotic disease in neoplastic patients]. PMID- 10686727 TI - [Umbilical metastasis of mucinous adenocarcinoma of the colon diagnosed by fine needle aspiration cytology]. PMID- 10686728 TI - Poverty and inequity: a proper focus for the new century. PMID- 10686729 TI - Health inequalities and the health of the poor: what do we know? What can we do? AB - The contents of this theme section of the Bulletin of the World Health Organization on "Inequalities in health" have two objectives: to present the initial findings from a new generation of research that has been undertaken in response to renewed concern for health inequalities; and to stimulate movement for action in order to correct the problems identified by this research. The research findings are presented in the five articles which follow. This Critical Reflection proposes two initial steps for the action needed to alleviate the problem; other suggestions are given by the participants in a Round Table discussion which is published after these articles. The theme section concludes with extracts from the classic writings of the nineteenth-century public health pioneer, William Farr, who is widely credited as one of the founders of the scientific study of health inequalities, together with a commentary. This Critical Reflection contributes to the discussion of the action needed by proposing two initial steps for action. That professionals who give very high priority to the distinct but related objectives of poverty alleviation, inequality reduction, and equity enhancement recognize that their shared concern for the distributional aspects of health policy is far more important than any differences that may divide them. That health policy goals, currently expressed as societal averages, be reformulated so that they point specifically to conditions among the poor and to poor-rich differences. For example, infant mortality rates among the poor or the differences in infant mortality between rich and poor sectors would be more useful indicators than the average infant mortality rates for the whole population. PMID- 10686730 TI - Socioeconomic inequalities in child mortality: comparisons across nine developing countries. AB - This paper generates and analyses survey data on inequalities in mortality among infants and children aged under five years by consumption in Brazil, Cote d'Ivoire, Ghana, Nepal, Nicaragua, Pakistan, the Philippines, South Africa, and Viet Nam. The data were obtained from the Living Standards Measurement Study and the Cebu Longitudinal Health and Nutrition Survey. Mortality rates were estimated directly where complete fertility histories were available and indirectly otherwise. Mortality distributions were compared between countries by means of concentration curves and concentration indices: dominance checks were carried out for all pairwise intercountry comparisons; standard errors were calculated for the concentration indices; and tests of intercountry differences in inequality were performed. PMID- 10686731 TI - Inequality of child mortality among ethnic groups in sub-Saharan Africa. AB - Accounts by journalists of wars in several countries of sub-Saharan Africa in the 1990s have raised concern that ethnic cleavages and overlapping religious and racial affiliations may widen the inequalities in health and survival among ethnic groups throughout the region, particularly among children. Paradoxically, there has been no systematic examination of ethnic inequality in child survival chances across countries in the region. This paper uses survey data collected in the 1990s in 11 countries (Central African Republic, Cote d'Ivoire, Ghana, Kenya, Mali, Namibia, Niger, Rwanda, Senegal, Uganda, and Zambia) to examine whether ethnic inequality in child mortality has been present and spreading in sub Saharan Africa since the 1980s. The focus was on one or two groups in each country which may have experienced distinct child health and survival chances, compared to the rest of the national population, as a result of their geographical location. The factors examined to explain potential child survival inequalities among ethnic groups included residence in the largest city, household economic conditions, educational attainment and nutritional status of the mothers, use of modern maternal and child health services including immunization, and patterns of fertility and migration. The results show remarkable consistency. In all 11 countries there were significant differentials between ethnic groups in the odds of dying during infancy or before the age of 5 years. Multivariate analysis shows that ethnic child mortality differences are closely linked with economic inequality in many countries, and perhaps with differential use of child health services in countries of the Sahel region. Strong and consistent results in this study support placing the notion of ethnicity at the forefront of theories and analyses of child mortality in Africa which incorporate social, and not purely epidemiological, considerations. Moreover, the typical advantage of relatively small, clearly defined ethnic groups, as compared to the majority in the national population, according to fundamental indicators of wellbeing--child survival, education, housing, and so forth--suggests that many countries in sub-Saharan Africa, despite their widespread poverty, are as marked by social inequality as are countries in other regions in the world. PMID- 10686732 TI - Defining and measuring health inequality: an approach based on the distribution of health expectancy. AB - This paper proposes an approach to conceptualizing and operationalizing the measurement of health inequality, defined as differences in health across individuals in the population. We propose that health is an intrinsic component of well-being and thus we should be concerned with inequality in health, whether or not it is correlated with inequality in other dimensions of well-being. In the measurement of health inequality, the complete range of fatal and non-fatal health outcomes should be incorporated. This notion is operationalized through the concept of healthy lifespan. Individual health expectancy is preferable, as a measurement, to individual healthy lifespan, since health expectancy excludes those differences in healthy lifespan that are simply due to chance. In other words, the quantity of interest for studying health inequality is the distribution of health expectancy across individuals in the population. The inequality of the distribution of health expectancy can be summarized by measures of individual/mean differences (differences between the individual and the mean of the population) or inter-individual differences. The exact form of the measure to summarize inequality depends on three normative choices. A firmer understanding of people's views on these normative choices will provide a basis for deliberating on a standard WHO measure of health inequality. PMID- 10686733 TI - Inequalities in health care use and expenditures: empirical data from eight developing countries and countries in transition. AB - This paper summarizes eight country studies of inequality in the health sector. The analyses use household data to examine the distribution of service use and health expenditures. Each study divides the population into "income" quintiles, estimated using consumption expenditures. The studies measure inequality in the use of and spending on health services. Richer groups are found to have a higher probability of obtaining care when sick, to be more likely to be seen by a doctor, and to have a higher probability of receiving medicines when they are ill, than the poorer groups. The richer also spend more in absolute terms on care. In several instances there are unexpected findings. There is no consistent pattern in the use of private providers. Richer households do not devote a consistently higher percentage of their consumption expenditures to health care. The analyses indicate that intuition concerning inequalities could result in misguided decisions. It would thus be worthwhile to measure inequality to inform policy-making. Additional research could be performed using a common methodology for the collection of data and applying more sophisticated analytical techniques. These analyses could be used to measure the impact of health policy changes on inequality. PMID- 10686734 TI - Public spending on health care in Africa: do the poor benefit? AB - Health care is a basic service essential in any effort to combat poverty, and is often subsidized with public funds to help achieve that aim. This paper examines public spending on curative health care in several African countries and finds that this spending favours mostly the better-off rather than the poor. It concludes that this targeting problem cannot be solved simply by adjusting the subsidy allocations. The constraints that prevent the poor from taking advantage of these services must also be addressed if the public subsidies are to be effective in reaching the poor. PMID- 10686735 TI - Round table discussion. Health inequalities and the health of the poor. PMID- 10686736 TI - Equality, equity: why bother? PMID- 10686737 TI - Overcoming inequity means finding approaches that work PMID- 10686738 TI - Combining forces against inequity and poverty rather than splitting hairs PMID- 10686739 TI - Efficient equity-oriented strategies for health PMID- 10686740 TI - Equity and gender PMID- 10686741 TI - Understanding and setting up the process for health equity PMID- 10686742 TI - The key to overcoming inequality is political commitment PMID- 10686743 TI - William Farr's legacy to the study of inequalities in health. AB - This section looks back to some of the ground-breaking contributions to public health, reproducing them in their original form and adding a commentary on their significance from a modern-day perspective. To complement this month's theme issue of the Bulletin, Margaret Whitehead assesses the importance of William Farr's contribution to the study of social inequalities in health. PMID- 10686744 TI - Cost-effectiveness of iron supplementation and malaria chemoprophylaxis in the prevention of anaemia and malaria among Tanzanian infants. AB - Prerequisites for effective interventions against severe anaemia and malaria among infants are economic evaluations to aid the setting of priorities and the making of health policy. In the present study we analysed the cost and effectiveness of three control strategies hypothetically delivered through the Expanded Programme on Immunization (EPI). For the prevention of severe anaemia and from the perspective of the health provider, the cost-effectiveness ratios were, respectively, US$ 8, US$ 9, and US$ 21 per disability-adjusted life year (DALY) for malaria chemoprophylaxis with Deltaprim (a combination of 3.125 mg pyrimethamine and 25 mg dapsone) + iron, Deltaprim alone, or iron supplementation alone. For malaria prevention, Deltaprim + iron cost US$ 9.7 per DALY and Deltaprim alone cost US$ 10.2 per DALY. From a sociocultural perspective the cost effectiveness ratios ranged from US$ 9 to US$ 26 for severe anaemia prevention and from US$ 11 to US$ 12 for the prevention of clinical malaria. These ratios were highly cost-effective, as defined by the World Bank's proposed threshold of less than US$ 25 per DALY for comparative assessments. Furthermore, all the preventive interventions were less costly than the current malaria and anaemia control strategies that rely on clinical case management. This economic analysis supports the inclusion of both malaria chemoprophylaxis and iron supplementation delivered through EPI as part of the control strategies for these major killers of infants in parts of sub-Saharan Africa. PMID- 10686745 TI - Deterioration in the nutritional status of young children and their mothers in Brazzaville, Congo, following the 1994 devaluation of the CFA franc. AB - The effects of the January 1994 devaluation of the African Financial Community (CFA) franc on the nutritional situation of the populations concerned has been little documented. We report in this article on two nutritional cross-sectional surveys that were conducted before and after this devaluation (1993 and 1996) in two districts of Brazzaville, Congo. The surveys involved a representative sample of 4206 households with a child aged 4-23 months. Complementary feeding practices and the anthropometric indices of the children and their mothers were compared, adjusting for changes in household socioeconomic characteristics. The results show a decline in the quality of the first complementary foods offered to the infants, i.e. less frequent use of special transitional foods and imported complementary flours (of higher nutritional quality), and preparation of less nutritious local gruels. Overall, the nutritional situation had deteriorated, with greater levels of stunting and wasting among children, mothers with lower body mass index, and infants with reduced birth weights. Increased food prices would appear to be the direct cause of the decreased quality in complementary feeding, but factors other than the devaluation have also had an impact on household welfare. The influence of these factors on nutritional-status is discussed. PMID- 10686746 TI - Detection of trypanosomes in suspected sleeping sickness patients in Uganda using the polymerase chain reaction. AB - Diagnosis of sleeping sickness (trypanosomiasis) is difficult because of the fluctuating levels of parasitaemia encountered in patients. In the present study we found that the polymerase chain reaction (PCR) demonstrated trypanosome infection in 20 out of 35 (57.1%) blood samples and in 21 out of 34 (61.7%) cerebrospinal fluid (CSF) samples collected from an area endemic for sleeping sickness in north-west Uganda. A total of 14 blood samples and 13 CSF samples that were positive for trypanosomes by double centrifugation were also positive by PCR, demonstrating good concordance between the two methods. However, 6 (28.6%) of the 21 blood samples that were parasitologically negative were positive by PCR, while 8 (38.0%) out of 21 CSF samples that were negative by double centrifugation were positive by PCR. These 14 negative samples could therefore be from sleeping sickness cases even though a positive PCR test is not evidence for the presence of trypanosomes. Furthermore, of these 8 CSF samples, 4 had been designated as early cases, based on the absence of trypanosomes and on a count of < or = 5 white blood cells (WBC) per microliter. This suggests that some late-stage cases could potentially be missed according to the present criteria, and it is therefore important to perform clinical trials to determine whether these cases could be treated successfully with the first-stage drug alone. The remaining four CSF samples had been classified as late-stage cases, based on a count of > 6 WBC per microliter, even though trypanosomes could not be detected in these samples by either double centrifugation or PCR. A cut-off point of 5 WBC per microliter, which is used as a rule of thumb to stage sleeping sickness patients, seems to leave some late-stage cases undetected since trypanosomes were detected in four CSF samples from suspected cases with < 5 WBC per microliter. PMID- 10686747 TI - Health system reform and the role of field sites based upon demographic and health surveillance. AB - Field sites for demographic and health surveillance have made well-recognized contributions to the evaluation of new or untested interventions, largely through efficacy trials involving new technologies or the delivery of selected services, e.g. vaccines, oral rehydration therapy and alternative contraceptive methods. Their role in health system reform, whether national or international, has, however, proved considerably more limited. The present article explores the characteristics and defining features of such field sites in low-income and middle-income countries and argues that many currently active sites have a largely untapped potential for contributing substantially to national and subnational health development. Since the populations covered by these sites often correspond with the boundaries of districts or subdistricts, the strategic use of information generated by demographic surveillance can inform the decentralization efforts of national and provincial health authorities. Among the areas of particular importance are the following: making population-based information available and providing an information resource; evaluating programmes and interventions; and developing applications to policy and practice. The question is posed as to whether their potential contribution to health system reform justifies arguing for adaptations to these field sites and expanded investment in them. PMID- 10686748 TI - Anthropological perspectives on injections: a review. AB - Qualitative studies from developing countries have pointed to the widespread popularity of injections. In addition to their use by formal and informal providers and traditional healers, there is now increasing evidence of the use of injections and injection equipment by lay people. Epidemiological research links the large number of unsafe injections to serious bloodborne infections such as viral hepatitis B and C and acquired immunodeficiency syndrome (AIDS). The present article examines the reasons behind the demand for injections by consumers and the administration of unnecessary or unsafe injections by different types of provider. Interventions aimed at reducing the risk of unsafe injections are discussed in relation to cultural and social factors as well as those factors associated with health systems. Suggestions are made for approaches to the design of such interventions. PMID- 10686749 TI - A new paradox: drugs too cheap to stay available. PMID- 10686750 TI - Unified terminology for pathology of the cervix. PMID- 10686751 TI - Toward a lexicon of population health. AB - Despite its undeniable currency in research and policy circles, there remains considerable confusion about what 'population health' is. We propose a lexicon for population health in the hope of clarifying issues and advancing this important research emphasis and policy agenda. It distinguishes population health in its literal meaning from a population health perspective, population health research, a population health framework, and a population health approach to policy. Population health is more than just thinking in aggregate terms or about identifying vulnerable or at-risk subpopulations. A population health perspective is fundamentally concerned with the social nature of health influences. The social structures that shape health experiences transcend the characteristics or actions of any one individual, providing population health with analytic advantages over individualistic-oriented approaches to health and to health policy. PMID- 10686752 TI - Collective lifestyles as the target for health promotion. AB - The last five years have witnessed intense debate among health researchers in Canada regarding the overlap of the health promotion and population health discourses. Meanwhile, strong currents within health promotion have attempted to move the field beyond a focus on individual behaviour towards the influence of social environments on health, although the tendency is often to fall back on individual behaviour modification as the primary lever for change. The Population Health research agenda bypasses behavioural determinants of health and explores instead social determinants. This body of knowledge provides useful insight for addressing some of the tensions in the health promotion discourse. This paper explores two of these tensions: whether individuals at risk or general populations should be targeted for change; and whether lifestyle is an individual or a collective attribute. We propose the notion of collective lifestyles as a heuristic for understanding the interaction between social conditions and behaviour in shaping health. PMID- 10686753 TI - Population health promotion: responsible sharing of future directions. AB - Population health promotion illustrates most robustly that health is a shared responsibility. Improving our understanding of the social production of health and the purchase population health promotion has on shaping social welfare policy presents a number of challenges to the future development of this discourse. Three are briefly discussed in this paper. First is the matter of language we use to describe our understanding of processes and influences. Second is the conceptualization of the pathways that shape population health status. Finally, cultural practices both extant and required to improve health status and reduce inequalities are addressed. PMID- 10686754 TI - Different wor(l)ds: three approaches to health research. AB - This article describes three approaches to research in the social sciences: positivist, interpretivist and critical social science. It uncovers some of the philosophical assumptions these approaches adhere to and situates the discussion in the population health arena with respect to these assumptions. The issues under debate are as yet unsolved (and perhaps unsolvable), with long histories in philosophy and sophisticated rationales on all sides. The article advocates defining the underlying terms of discussion and making assumptions explicit to facilitate dialogue, and also encourages exploration of and tolerance for other approaches. PMID- 10686755 TI - Indicators that count! Measuring population health at the community level. AB - We begin with a discussion of some vitally important conceptual and methodological issues. These issues concern our understanding of community, of health, of population health and its determinants, of the concept of 'measurement' and the values that underlie it, and our reasons for wishing to measure these constructs. We then present a framework for indicator categories, propose some criteria for indicator selection and suggest an initial set of core indicators. This indicator set reflects not simply health status--no matter how broadly defined--but also the environmental, social and economic determinants of health and the "healthfulness" of the community itself. Our most important conclusion is that if the information that is contained in the data of the indicator set is to be transformed into knowledge that can empower and emancipate the community, it has to be developed in consultation with the local community and local users of the information. PMID- 10686756 TI - From concept to practice: including the social determinants of health in environmental assessments. AB - The present paper examines the historical evolution of health impact assessments as part of the environmental assessment process. The development of a coherent public health framework must be based on the model of determinants of health, integrating toxic and infectious risks and social impacts of projects. The integration of common concepts, processes and methodologies from the area of public health and social impact assessment challenges the quantitative model approach to risk assessment. The expert-driven risk assessment is transformed into a social learning process where local knowledge and scientific input foster a dialogue among stakeholders. The issue-oriented, iterative and participative assessment process may be applied to the health impact assessment of public policies. Sustainable development with its social objectives of empowerment, participation, equity, poverty alleviation, social cohesion, population stability and institutional development is an appropriate framework for conducting health impact assessments. PMID- 10686757 TI - Health, environmental assessments and population health: tools for a complex process. AB - Place is more than physical and natural environment. The role of biophysical environment has still to be articulated in population health discourse and its relations with human health are fraught with scientific uncertainty and dissension. An environmental impact assessment (EA) evaluates the environmental effects of a proposal--a rational and technical process. Sometimes health assessments are included, usually by quantitative risk assessments which are subject to the limits of scientific knowledge and bedevilled by data limitations. The goal must be to add health to the process, yet the relevant features to include are complex. Impacts are non-specific and they interact and have spatial and temporal characteristics. To integrate environment into population health, there is a need for a physical environment-health database and inter-sectorial policy and action. There is also a need for different types of indicators to measure process, impact and effectiveness, and for new tools (stories, photography) to account for context and values. PMID- 10686758 TI - British Columbia's health reform: "new directions" and accountability. AB - The health policy New Directions committed the British Columbia government to a population health perspective and extensive community involvement in the health services reform process. The policy envisaged elected citizen boards with authority to raise revenues and exercise a significant degree of local autonomy. Academic and public attention has been paid to the decision in November 1996 to collapse New Directions' two-tier governance structure into a single level. Less attention has been paid to the profound changes that occurred prior to the government's reversal on the question of governance. This paper focuses on those changes. During the critical three years between the 1993 launch of the reform and its formal revision in 1996, the government's positions on elections, taxation power, local autonomy and scope of action for regional boards all changed. Those changes marked a retreat from political accountability to the community and an advance towards managerial accountability to the government. PMID- 10686759 TI - Factors that facilitated and challenged the development of health goals and targets: the British Columbia experience. AB - Health promotion research and practice reveal that goal setting and monitoring have gained increased acceptance at international, national, provincial/state, regional and local levels. The global adoption of health goals as a strategy for population health promotion has occurred even though few protocols or guidelines to support the health goals development process have been published. Limited study has occurred on the variation in approach to health goals planning, or on the complex, multiple forces that influence the development process. This paper describes conclusions drawn from an exploratory and descriptive case study that tracked the pathways to health goals in British Columbia (BC) and uncovered nearly 100 factors that influenced the final version of health goals adopted by the government of BC. Influencing factors included: (a) positive perceptions of the benefits of health goals, (b) the role of a trusted health goals champion, (c) positioning of the goals as government rather than health ministry goals, (d) the format and agenda of the health goals consulting process, and (e) political reluctance toward highly specific or measurable goals with targets. PMID- 10686760 TI - Achieving population health goals: perspectives on measurement and implementation from Australia. AB - Health goals and targets have been widely used to indicate strategic direction and priority for health improvement on a population basis. This paper provides an overview of Australia's experience in using health targets and considers the relevance of this experience for Canada. It gives special attention to the challenge of developing a broadly based set of targets that reflect the social, economic and environmental determinants of health alongside more traditional measures of health status. It examines how the technical challenge of measurement, the bureaucratic barriers between government departments, and the political conservatism inherent in federal systems of government present formidable barriers to effective action on comprehensive national health targets. The paper concludes with a reminder of the need for inter-sectorial action to address the determinants of health. Based on the Australian experience, it suggests for Canada an ideal combination of a national population health framework to guide direction and priority, to be implemented through action at a more local level, through well-defined partnerships. PMID- 10686761 TI - Understanding the determinants of health: key decision makers in Saskatchewan Health districts and Saskatchewan Health, 1998. AB - This research inquiry used qualitative and quantitative methods to examine how key decision makers from Saskatchewan health districts and Saskatchewan Health understand the determinants of health. The inquiry was based on the premise that key decision makers' understanding of the determinants of health, and the consensus regarding these understandings, hinder or facilitate dialogue, choice of effective strategies, and achievement of health promotion goals. Interviews indicated variation in perspective and emphasis regarding how key decision makers understand the determinants of health. A survey of key decision makers found: 1) inconsistencies in respondents' understanding of the determinants of health, particularly between stated beliefs and priorities for actions; and 2) that the degree of consensus among decision makers was higher for stated beliefs and lower for choices of action. Results indicate a need for clarification and consensus building processes concerning the determinants of health, as well as for clear policies that foster consistency between beliefs and actions and minimize inappropriate or undesirable differences in interpretations. PMID- 10686762 TI - The 'people assessing their health' (PATH) Project: tools for community health impact assessment. AB - The People Assessing Their Health (PATH) Project was designed to provide a means for people in selected communities within Eastern Nova Scotia to become more involved in decision making within the province's emerging decentralized health system. Using community health impact assessment (CHIA) as a population health strategy, community members were able to identify factors that determine their health and to develop tools to help them assess the health impact of programs and policies within their communities. The participatory process used throughout the PATH Project enabled a wide range of people to generate information for designing a community health impact assessment tool (CHIAT) unique to their community. It also helped participants to broaden their understanding of the many factors determining health of their community and of the region. PMID- 10686763 TI - Health indicator development in Alberta health authorities: searching for common ground. AB - The ability to measure population health trends and improvements can be enhanced through collaborative efforts to describe existing knowledge and via shared development opportunities. This paper highlights a project undertaken in Alberta which has created an inventory of health status indicators in use in the province, and provides a framework for strategic progress in the development and use of a common set of indicators across the province. The work may provide a model for other regional health authorities interested in comparing the health of their populations across time and across health regions. PMID- 10686764 TI - Applying a population health approach. AB - While concepts that underlie good public health and population approaches to health go back a long way, renewed recognition that health is dependent on more than the ability to treat has given new impetus to a more comprehensive approach to thinking about and planning for health and human services. This paper offers a reflection on how we conceptualize population approaches to health. Recognizing our current understanding of health determinants and dynamics, the paper explores moving from "avoiding disease" to to "pursuing health." It then examines the pragmatic balancing act of science, art, beliefs and politics, with attendant traps. It concludes with a way of framing action on population health and translating theory into practice. PMID- 10686765 TI - Shared responsibility for population health: a personal reflection. PMID- 10686766 TI - Advancing the population health agenda: uniting altruism and self-interest. PMID- 10686767 TI - Future directions in population health. AB - The long-term health of the population will be influenced by a number of major forces in the next century. In this brief review, particular emphasis is placed on environmental and economic forces. Major global environmental changes include climate change and global warming, resource depletion, ecotoxicity and reduced biodiversity. We do not yet know the impact on longevity of lifetime exposure to a mix of persistent toxic chemicals in our environment, since it has only been widespread in the past 40-50 years. The health impacts of global warming are only just beginning to be understood and could be profound. But perhaps the most profound threat to population health is economic growth, to the extent that it undermines environmental and social sustainability. We need a new form of capitalism, one that simultaneously increases environmental, social, economic and human capital, if population health is to be maintained in the 21st century. PMID- 10686768 TI - Population health in Canada: issues and challenges for policy, practice and research. AB - The population health movement has gained prominence in Canada and elsewhere with policy makers, program planners and researchers taking note that health is strongly influenced by factors that lie largely beyond the health-care system. The development of population health in Canada was the focus of the National Conference on Shared Responsibility for Health & Social Impact Assessments: Advancing the Agenda held May 2-3 1999 in Vancouver, Canada. A longer version of this paper was distributed to conference participants to provide some common knowledge and vocabulary. It also introduced and discussed definitional, normative, logistical, political, methodological, structural and resource considerations with respect to furthering the population health agenda in Canada. PMID- 10686769 TI - Endovascular treatment of aortic aneurysms of the abdominal aorta with covered stents. AB - Abdominal aortic aneurysms are common in the aging population; their surgical treatment is well established and allows good results in specialized centers. Endovascular exclusion of abdominal aortic aneurysms has been shown to be feasible since 1991 and nowadays commercially available bifurcated endografts allow safe exclusion in selected cases. In the last year 22 patients with an aorto-iliac aneurysm received endovascular treatment at our Institution. We included patients with favorable anatomic characteristics (i.e. neck > 15 mm length, and < 28 mm diameter, iliac neck < 12 mm diameter, absence of > 90 degrees iliac or aortic angulation) and, in particular, those with increased surgical risk for systemic pathology (12 patients), or hostile abdomen (9 patients). We employed Vanguard II (Boston Scientific) endovascular grafts introduced through a surgically exposed common femoral artery; the contralateral limb of bifurcated grafts was inserted percutaneously. The endograft was successfully implanted in all cases, requiring additional iliac cuffs for complete aneurysm exclusion in 3 cases. Periprocedural morbidity included one case of thrombosis and one case of pseudoaneurysm of the punctured femoral artery, which required surgical treatment. In one case surgical exposure of the iliac artery was required in order to advance the device into the aorta. In one patient who previously underwent hemicolectomy, postoperative colonic ischemia was observed, and pharmacological treatment was required. Moreover we also observed one case of groin infection that was treated successfully with local wound care and systemic antibiotics, and one late contralateral limb thrombosis that was successfully treated with loco-regional thrombolysis. The mean follow-up was 6.1 months: one patient died because of congestive heart failure. No further morbidity was recorded. A type-II endoleak was observed in one patient, originating from the inferior mesenteric artery with no sac enlargement; this patient is still under observation. In conclusion, with proper clinical selection, commercially available endovascular devices allow safe exclusion of abdominal aortic aneurysms. Long-term follow-up is needed to ascertain the durability of the procedure. PMID- 10686770 TI - Cardiac risk stratification of patients undergoing peripheral vascular surgery. AB - Approximately 50% of patients with peripheral vascular disease have severe coronary artery disease. Several ways of predicting the postoperative risk of major cardiac events in peripheral vascular patients have been suggested. Among preoperative tests, echocardiography is now receiving greater favor for risk stratification. PMID- 10686771 TI - Preinfarction angina and myocardial preconditioning. AB - In the current era of pharmacologic and mechanical reperfusion therapy, several studies have consistently shown that patients with myocardial infarction preceded by angina have smaller infarcts and a better in-hospital outcome after thrombolytic therapy than patients without preinfarction angina. At least three mechanisms may explain these differences between infarctions that are preceded by angina and those that are not: coronary collaterals, reperfusion rate, and ischemic preconditioning. Collaterals alone do not seem to explain the beneficial effects of preinfarction angina, although it is difficult to completely rule out their role in the clinical setting. The possibility that preinfarction angina is not protective per se, but rather is a predictor of a more rapid coronary reperfusion is attractive; however, it should be addressed by further clinical studies. Finally, it is likely that the beneficial effects of preinfarction angina are related to ischemic preconditioning. Although a direct demonstration of this hypothesis is still lacking, clinical features of preinfarction angina, which is characterized by anginal attacks preceding acute myocardial infarction, are very similar to those of ischemic preconditioning, in which brief ischemic episodes precede a prolonged ischemic period. Indeed, the demonstration of ischemic preconditioning in different clinical models of ischemia and reperfusion and the identification of some of its mediators suggest that in patients at high risk of myocardial infarction drugs known to block this endogenous form of protection should be used with caution, while drugs known to elicit preconditioning might have a relevant therapeutic role. However, the optimal timing, administration, and dosage for preconditioning-mimetic drugs in the appropriate clinical setting are still under debate and warrant further investigation. PMID- 10686772 TI - Emerging antithrombotic treatments for acute coronary syndromes. AB - In the last few years the hypothesis of coronary thrombosis, frequently triggered by plaque ulceration or fissuration, has gained wide acceptance as one of the key events in the pathophysiology of acute coronary syndromes. Plaque ulceration may activate both platelets and the coagulation cascade via exposure of a variety of substances, such as von Willebrand factor and tissue factor. It has been demonstrated that aspirin reduces mortality and improves the prognosis of patients with such syndromes. More recently, newer drugs have been identified for the treatment of acute coronary syndromes; in particular, platelet glycoprotein IIb/IIIa inhibitors have been found to be more effective than aspirin in a variety of clinical conditions, such as unstable angina, acute myocardial infarction, and coronary angioplasty. Other drugs with different mechanisms of action will be soon available for large scale clinical trials. PMID- 10686773 TI - Rescue coronary angioplasty: a crotch for limping thrombolysis? AB - After failed thrombolysis, rescue coronary angioplasty is performed with the aim of restoring complete flow in the infarct-related artery. The clinical benefit of this strategy has been debated in few clinical trials during the early '90s, and high procedure-related risks, low success and early reocclusion rates seemed to outweigh the benefit of mechanical recanalization. The RESCUE trial and, more recently, data from the GUSTO angiographic substudy supported the hypothesis of a better outcome among patients aggressively managed after failed thrombolysis. Noninvasive identification of such patients must be accomplished monitoring electrocardiogram and biochemical markers of myocardial necrosis. Further improvements in the management of candidates to rescue coronary angioplasty can be obtained with a more liberal use of intra-aortic balloon pump among subjects admitted in cardiogenic shock; stents and platelet aggregation inhibitors could reduce early reocclusion, but randomized data are needed to test this hypothesis. PMID- 10686774 TI - Oxygen consumption. AB - It gets more and more frequent to use oxygen consumption (VO2) to evaluate exercise capacity and response to treatment in heart failure patients. The amount of VO2 is due to ventilation, oxygen transport and muscle activity. No one of these single steps can define by itself VO2, but all these physiological functions are integrated each other. In this paper we examine the modifications of cardiac output, arteriovenous oxygen content difference, and the temporal behavior of their variations during exercise in heart failure. We specifically describe changes in VO2 during simulated altitude; we also contemplate mechanisms governing oxygen diffusion from capillary bed to mitochondria and critical capillary PO2 concept. PMID- 10686775 TI - Short-term effect of exercise on platelet factor 4 in normal subjects and in patients with coronary artery disease. AB - BACKGROUND: The aim of this study was to evaluate the plasma concentration of a platelet-derived protein, platelet factor 4 (PF4), before and after exercise in coronary artery disease. METHODS: We enrolled 60 patients with documented ischemic heart disease. The subjects were divided into two groups: Group 1 patients with previous myocardial infarction (n = 20, 13 males, 7 females, mean age 51.6 +/- 7.5 years, range 38-62 years); Group 2 patients with exercise induced angina (n = 40, 22 males, 18 females, mean age 52.6 +/- 8.0 years, range 38-65 years). Patients with hypertension, hyperlipidemia and diabetes were excluded. Patients with angina or ST segment depression during the stress test were included in a subgroup (n = 33, 21 males, 12 females, mean age 50.3 +/- 6.3 years, range 40-65 years). Twenty healthy subjects without coronary risk factors (13 males, 7 females, mean age 53.2 +/- 7.1 years, range 38-65 years) served as controls. PF4 was measured in all patients at baseline and 5 min after a bicycle exercise test. Plasma PF4 levels were measured performed by radioimmunoassay (ng/ml, normal range 0-10). RESULTS: Patients with ischemic heart disease showed a high basal concentration of PF4 compared with controls. PF4 levels at baseline vs after stress test were 4.1 +/- 2.5 vs 5.3 +/- 2.6 ng/ml in healthy subjects; 33.4 +/- 15.8 vs 56.2 +/- 28.2 ng/ml (p < 0.001) in Group 1; 22.4 +/- 15.8 vs 44.6 +/- 28.4 ng/ml (p < 0.001) in Group 2; 29.9 +/- 15.5 vs 67.7 +/- 26.1 ng/ml in the subgroup with angina or ST segment depression (p < 0.001), and 23.1 +/- 16.5 vs 26.0 +/- 18.1 ng/ml in those without angina or ST segment depression (NS). CONCLUSIONS: These findings support the hypothesis that a significant increase in PF4 levels after exercise is associated with clinically significant coronary artery disease. PMID- 10686776 TI - Platelet activation with exercise in coronary disease. Is it ischemia or atherosclerosis? PMID- 10686777 TI - Left ventricular metastasis from uterine leiomyosarcoma. AB - Cardiac metastases are uncommon but seem to be increasing in incidence, possibly in relation to prolonged survival of cancer patients. Leiomyosarcoma metastatic to the heart is extremely rare. We report the case of a 57-year-old woman previously treated for uterine leiomyosarcoma who presented with dyspnea, electrocardiographic changes mimicking myocardial infarction, and normal enzymes. A left intraventricular mass, suspected as cardiac metastasis, was revealed by echocardiography. The patient died 1 week later. At autopsy the mass proved to be histologically a metastasis of the uterine tumor. PMID- 10686778 TI - [50 years among the pioneers of a new medical specialty; the development of modern traumatology]. AB - On the basis of his half a century medical experiences the author analyses how the traumatology became an independent branch of medicine.--L. Markusovszky's scientific achievements in the domain of the field surgery, organization, education and professional policy considerably influenced the development of the traumatology. Derived from the surgery, the present-day traumatology's basis feature is to provide a high-level professional education and a well organised continuing medical attendance.--The achievements of the last fifty years are now disclosed to the readers on the strength of the experiences of the author and his contemporary collegues. He breaks the national events into 10 years periods which are in fact representing the efforts and the results of the European shools. Finally in the approach of the second millennium he is looking toward the future. With the full knowledge of the nowadays difficulties he is trying to answer how will advance all that has been founded in the last fifty years. Especially as in the next few decades the harmonization European, an internationally similar high quality medical attendance will be a considerable task. PMID- 10686779 TI - [The role of alpha-interferon therapy in chronic myeloid leukemia]. AB - Alpha-interferons are widely accepted and used in chronic myeloid leukaemia. The standard indication is the chronic phase, and it seems to be clear, that by prolonging the chronic phase interferon therapy results in better overall survival than busulphan or hydroxyurea (this comparison needs metaanalysis) monotherapy. A review and recommendation is given considering the indications (with a special attention to stemcell transplantation), dosage, contraindications and monitoring interferon treatment. PMID- 10686780 TI - [Ovulation induction therapy and malignant ovarian cancer]. AB - In the authors' institute 1097 patients received treatment for ovarian cancer between the 1st of January, 1960 and the 31st of December, 1997. 92 of them had malignant granulosa cell tumor. In this study the link between ovulation induction therapy and ovarian cancer was analyzed with retrospective questionnaire. 236 questionnaires were shared out among patients with malignant ovarian tumor, who were treated between 1990 and 1997. 7 of 113 patients, who gave correct answers to the questions (6.2%) received ovulation induction therapy. Epithelial ovarian cancer developed in 2 of the cases during, and in 5 cases just 6-16 years after the clomiphene-citrate treatment. None of the 45 patients with granulosa cell tumors received induction therapy. The number of patients admitted because of malignant ovarian tumor before and after the induction therapy was also compared. There was no significant increase in the occurrence of the malignancy. Since 1986 in the in vitro fertilization program of the clinic nearly 1,500 patients were treated with effected ovulation induction drugs causing superovulation. The authors don't know of any development of malignant ovarian tumor, and 732 woman have confirmed this fact. The number of patients deceased in consequence of ovarian cancer in different age groups, and the distribution of the women population in every age group in Hungary, from the year of 1979 was also analyzed. There was no significant increase found in the number of deceased among the studied age groups, moreover a significant decrease among them could be observed. The link between ovulation induction therapy and ovarian cancer can neither be strengthen, nor deny by the result of the study, however the close relationship which seemed to be logical can be queried. To give an exact answer for the question a vast, long-term, prospective of retrospective follow-up case-control study is needed. PMID- 10686781 TI - [Use of two teflon stents in malignant non-hilar biliary atresia]. AB - Insertion of biliary stents in cases of malignant biliary obstruction is a widely accepted method to resolve jaundice. The authors applied two 10 French biliary teflon stents to prolong the drain patency in 32 patients with distal malignant obstruction, thought to be inoperable at the time of intervention. Among the followed, inoperable 23 cases 14 patients died for the time of evaluation. The median survival was 150 days, the median drain patency was 99 days. Cholangitis was the cause of death in three patients. Repeated endoscopic interventions were: transient nasobiliary drainage without drain replacement in two patients and four changes of stents in three patients. In the 13 patients, surviving and wearing their drains at least for 100 days the patency of the double drains was 157 days. These results obtained in the long-time survivors support the comparability of the patency of double teflon stent to that of metalstents. In majority of cases the two teflon drains remained patent until the deaths of patients. PMID- 10686782 TI - [Beginning of the cholera epidemic in 1848]. PMID- 10686783 TI - [Elections in Hungary--Physicians and pharmacists in the independent government 1861-1862]. PMID- 10686784 TI - [Commemorative coins for researchers in geriatrics and caregivers for the aged]. PMID- 10686785 TI - [Management of Lyme disease]. PMID- 10686786 TI - [Allergen immunotherapy--therapeutic vaccines for allergy and asthma]. PMID- 10686787 TI - [50 years PPmP--what lies ahead?]. PMID- 10686788 TI - [Using integrative approaches in psychotherapy]. AB - Surveys on both sides of the Atlantic show that eclecticism and integrative approaches in psychotherapy enjoy growing popularity. At the same time the idea of integration still meets with major resistance in schools of psychotherapy and training institutions, as well as in clinics. Therefore it not seems the right time to inquire into the usefulness of eclectic and integrative approaches. After a conceptual clarification this paper looks for criteria for the usefulness of such approaches and for empirical work establishing it, as well as for the conditions needed for such approaches to be useful. It is concluded that there are several reasons for, and hints at the usefulness of integrative approaches, but not yet many empirical validations. Recent empirical research, however, clearly shows that for certain patients and certain disorders and problems combined treatments are the treatments of choice, and that further research as well as better training in differential diagnostics and in combined treatments are not only of major interest to the patient and the therapists but are also feasible. PMID- 10686789 TI - [First all-Germany standardization of the brief form of the Gissen Complaints Questionnaire GBB-24]. AB - At the end of 1994 a shortened version of the Giessen Subjective Complaints List (GBB-24), was standardised using a sample of 2182 subjects aged 18 to 60 years, representative of the population of re-unified Germany (720 from East Germany, 1462 from West-Germany). The item norms are given for the whole group, while the scale norms are also subdivided according to gender, age (18-30, 31-40, 41-50 and 51-60 years) and place of residence (East/West). The dependence of physical complaints on age and gender has diminished significantly since 1975, whereas the factor structure and the internal consistency of the scales have changed only slightly. The relative influence of further socio-demographic variables (education, income, partnership status, town/country, unemployment) and attitudes health on the scale scores of the GBB-24 are described. The results were also correlated with data from Giessen-Test (GT) and a questionnaire on life satisfaction (Fragebogen zur Lebenszufriedenheit FLZ), collected at the same time. As expected, significant relationships were found between Scale 4 (Depression) of the Giessen-Test and all scales of the GBB-24, particularly Scale 1 "Exhaustion". A significant relationship with life satisfaction was also found, i.e. absence of complaints corresponded with greater life satisfaction. As anticipated, this relationship became most evident in the area of physical health. PMID- 10686790 TI - [Recent developments in systematic evaluation of the therapeutic process. Scales for the assessment of adherence and competence]. AB - The assessment of therapists' behaviour an essential part of psychotherapy research, especially for controlling the validity of studies. In the last decade, particularly in English-speaking countries, research has been developed that studied two constructs important in this context: adherence and competence. Adherence refers to the extent to which therapists use the strategies recommended in a therapy manual; competence refers to the quality of the use of these strategies. This article is a review of research associated with adherence and competence scales and its relevance. First of all, definitions and examples of use are presented and discussed. Secondly, some of the most important scales that have been developed in this context are described. Thirdly, aspects of realisation are presented. Finally, the pros and cons of this type of research are discussed. It is argued that despite some problems adherence and competence scales are a significant contribution to the systematic assessment of therapist behaviour. PMID- 10686791 TI - [Psychotherapists should at least be able to count to 3--or?]. PMID- 10686792 TI - [Methodological openness as the basis of integrative psychotherapy]. PMID- 10686793 TI - [Image of the month. Percutaneous closure of an interauricular communications]. PMID- 10686794 TI - [Pharma-clinics. How I treat ... HIV infection. III. Non-nucleoside reverse transcriptase inhibitors]. AB - Non nucleoside reverse transcriptase inhibitors (NNRTI) are a new arm in the treatment of the HIV infection. They inhibit the replication by direct non competitive binding to the enzyme, and do not require phosphorylation. The fast emergence of resistance in monotherapy obliges to use them in a triple association. The 103 mutation confers a cross-resistance. The most common adverse event is rash. Association with nucleoside analogues is additive or even synergistic. They are metabolized by the cytochrome P450. Within a combined therapy, their efficiency is comparable to protease inhibitors, notably in patients with low viral load. PMID- 10686795 TI - [Clinical case of the month. Mental confusion due to the administration of tramadol in a patient treated with MOAI]. AB - However currently less used, MAOI (monoamine-oxidase inhibitor) antidepressants have specific indications like refractory or atypical depressions. The main difficulties related to MAOI therapy consist in their potentially very dangerous interactions with certain foods on the one side, many medications on the other side. For example, we report the case of a patient treated for a resistant depression by phenelzine (Nardelzine) who presented a severe delirium after the administration of tramadol (Contramal, Dolzam). This case shows the importance of particular attention before the association of any drug in a patient treated by MAOI. PMID- 10686796 TI - [Therapeutic non-compliance: a major problem in the prevention of cardiovascular diseases]. AB - The prevention of cardiovascular diseases relies upon the correction of risk factors and, more particularly, the optimal management of various metabolic abnormalities such as obesity, dyslipidaemias, diabetes mellitus and arterial hypertension. Such an approach first requires the adherence to life-style habits (healthy diet, physical activity and no smoking) and, in case of failure, the use of lipid-lowering drugs, antidiabetic agents and/or antihypertensive medications. Sometimes, a monotherapy may be sufficient but, in most cases, a drug combination is mandatory because of the need to reach tight therapeutic targets and of the presence of a polypathology, especially within the frame of the metabolic syndrome. Unfortunately, all surveys indicate that therapeutic compliance to non pharmacological advice and even to drug prescriptions is far from being excellent. Such a non-compliance limits the efficacy of the prevention strategies and contributes to markedly increase the cost of metabolic diseases and associated complications. PMID- 10686797 TI - [Polyarteritis nodosa related to hepatitis b virus infection]. AB - A 66-year-old man was hospitalized for asthenia, weight loss, fever and chills. A polyarteritis nodosa associated with hepatitis B virus infection in a replicative phase was diagnosed. Etiology, physiopathology, and clinical characteristics of hepatitis B virus-related polyarteritis nodosa will be reviewed, as well as the effectiveness of therapy combining corticosteroids, interferon alpha and plasma exchanges. PMID- 10686798 TI - [Role of positron emission tomography is the evaluation of digestive tract tumors]. AB - Imaging and endoscopic techniques have taken an increasing part in the management of gastroenterological disorders. Among these techniques, FDG-PET imaging has emerged as a powerful tool in the management of several cancer diseases, including tumors of the digestive tract. In particular, the role of PET for diagnosing and staging recurrent colorectal cancers, and for differentiating mass forming pancreatitis from carcinoma is now well established. In this review, we will briefly discuss the place of PET imaging in the work-up of the tumors of the digestive tract. PMID- 10686799 TI - [Do you have a good nose? Small lexicon of bromhidroses and other body odors]. AB - Body odours have a semiological significance in dermatology and in internal medicine and pediatrics as well. Most of them are brought by the eccrine or apocrine sweat. The term bromhidrosis refers to such conditions. Body odours originate from metabolic disorders, ingestion of food, toxic elements and drugs, and from contact with various xenobiotics. Specific bacterial infections and contaminations also release volatile smelling compounds without the intervention of sweat. PMID- 10686800 TI - [Fibromuscular dysplasia]. AB - Fibromuscular dysplasia is a rare non-atherosclerotic, non-inflammatory arterial disease. It concerns less than 1% of all occlusive artery lesions, but is more common in young female patients, with a prevalence of 3 to 5% of the arterial lesions in that age group. It mainly attains renal and carotid arteries. The authors discuss the etiopathogeny, the prevalence and treatment of fibromuscular dysplasia. Their own surgical experience with 21 renal and 10 carotid lesions of fibromuscular is exposed. PMID- 10686801 TI - [How I investigate ... dyskalemia]. AB - Potassium is the most important intracellular cation, playing a role in neuromuscular excitability. Dyskalemia is common. The consequence of this ionic perturbation could be serious related to its magnitude. Thus a quick etiological approach and a medical intervention are needed to rapidly correct this potentially lethal ionic disturbance and prevent its recurrence. PMID- 10686802 TI - [Pharma-clinics. The drug of the month. Nevirapine. Viramune]. AB - Nevirapine is the first non nucleoside reverse transcriptase inhibitor registered in Belgium and indicated in the treatment of HIV-1 infection. In association with 2 nucleoside analogues, its efficiency is similar to a tritherapy with protease inhibitor, particularly in naive patients with low viral load. It has a good tolerance profile and is easy to take. Studies in progress should permit to widen its indications. PMID- 10686803 TI - [Clinical study of the month. The ATLAS study]. AB - The Atlas Study was set up to compare the efficacy and safety of low doses and high doses of ACE inhibition by lisinopril on the risk of death and hospitalization in chronic heart failure. Three thousand one hundred sixty-four patients with class II to IV heart failure and an ejection fraction below 30% were randomly assigned to double blind treatment with either low doses (2.5-5 mg/daily, n = 1596) or high doses (32.5-35 mg/daily, n = 1568) of the ACE inhibitor lisinopril for 39 to 58 months while background therapy for heart failure was continued. Patients in the high dose group had a non significant 8% lower risk of death (p = 0.128), but a significant 12% lower risk of death or hospitalizations for any reason (p = 0.002) and 24% fewer hospitalizations for heart failure (p = 0.002). Side-effects such as dizziness and renal insufficiency were more frequently encountered in the high dose group, but there was no difference between the two groups in terms of number of patients requiring discontinuation of study medication. These findings indicate that patients with heart failure should not, as too frequently is, be maintained on very low dose of an ACE inhibitor unless this is the only dose that can be tolerated. The patients are expected to benefit more if they receive higher doses close to those used in the large clinical trials which have demonstrated a reduction by ACE inhibition in morbidity and mortality in heart failure. PMID- 10686804 TI - [What is your diagnosis? Extragenital endometriosis in the left rectus abdominis muscle]. PMID- 10686805 TI - [Emerging and re-emerging infectious diseases--a challenge for public health]. AB - We have recently seen a worldwide explosion of infectious diseases: emerging diseases like the HIV/AIDS pandemic, or old diseases like cholera, tuberculosis, diphteria, plague, yellow fever, dengue, or malaria. These reemerging diseases are on the surge because of multiple factors: environmental changes, transformation of ecosystems, ongoing socioeconomic degradation and deterioration of public health systems in many countries. The increasing bacterial resistance to antibiotics or virologic resistance to antiviral drugs are becoming a serious problem today. This global danger needs a global response. There must be a cooperation between the different actors in the field of public health. The general practitioner should look for good therapeutic compliance, control vaccinations, and give his patients health education, including prevention of HIV and other sexually transmitted diseases. PMID- 10686806 TI - [Differential diagnosis and therapy of secondary hypertension (with special reference to renal artery stenosis) in general practice]. AB - Secondary hypertension is present in 3-5% of patients suffering from high blood pressure. Some of the causes of secondary hypertension can be cured and all of them need specific treatment. This can only be provided when an accurate diagnosis is sought. On the other hand, adequate knowledge of the different etiologies and their respective clinical presentation avoids unnecessary and expensive laboratory and imaging investigations. Renal artery stenosis is increasingly being recognised as a relevant and frequent cause of secondary hypertension and renal insufficiency, this previously underdiagnosed entity is thus covered in some detail. Endocrine causes of secondary hypertension are discussed, their diagnosis and treatment should be discussed with an endocrinologist. Before diagnosing hypertension as being refractory to therapy, adequate treatment and patient-compliance have to be enforced. Some frequently overseen but important causes of hypertension in daily practice are also discussed (drug-induced, loss of vessel wall elasticity in old age, etc.). PMID- 10686807 TI - [Differential malignant lymphoproliferative syndrome (LPS) diagnosis with flow cytometry: a study of 100 patients]. AB - The clinical utility of flow cytometry in diagnosis of chronic lymphoproliferative disorders (LPD) is well established. Accurate diagnosis of related but still distinct entities is relevant to therapeutic decisions. We report on the immunophenotypic findings of 100 patients with a new diagnosis of LPD established by two-color flow cytometry. A panel of > 15 monoclonal antibodies was regularly applied. The characteristic immunophenotype of B-cell chronic lymphocytic leukemia (CD5+, CD23+, FMC7-) was found in 74 patients including one with Richter's transformation. Hairy cell leukemia (CD5-, CD11c+, CD103+) was diagnosed in 6, and B-cell Non-Hodgkin lymphoma (B-NHL) in 13 patients, respectively. 6 of the B-NHLs belonged to the entity of splenic lymphoma with villous lymphocytes (CD5-, CD23-, FMC7+) and 6 were identified as mantle cell lymphomas (CD5+, CD23-, FMC7+). One B-NHL was typed as follicular center cell lymphoma (CD5-, CD10+, FMC7+). Three B-cell LPDs without a characteristic marker profile were histologically further classifiable. With a total of 4 patients T-cell LPDs were much less frequent. Sezary syndrome (CD4+, CD8-, CD56-) and T gamma lymphoproliferation (CD4-, CD8+, CD16-, CD56-, CD57+) were diagnosed twice. In 17 patients with a characteristic marker profile (1 Richter's transformation, 5 hairy cell leukemias, 3 splenic lymphomas with villous lymphocytes, 4 mantle cell lymphomas, 4 T-cell proliferations) a further histological or molecular investigation confirmed the immunophenotypic diagnosis in all cases. Clinical presentation with lymphadenopathies and B-symptoms was mainly associated with the diagnosis of mantle cell lymphoma, whereas splenomegaly and infection were suggestive of hairy cell leukemia. 94% of the B CLL patients were diagnosed at an early clinical stage with still conserved hematopoiesis, 32% of the LPDs were diagnosed following a routine hematogramm. PMID- 10686808 TI - [Treatment concept in glaucoma]. PMID- 10686809 TI - [What is your diagnosis? Adenoid cystic prostatic carcinoma]. PMID- 10686810 TI - [Prevalence of diagnoses in ambulatory and hospitalized patients]. AB - Prevalence and distribution of different clinical presentations were analyzed in 6000 hospitalized and ambulatory patients. One medical University clinic and three further hospital units, two university outpatient clinics and four practitioners participated. 6000 main diagnoses and 2560 additional diagnoses had been recorded. The mean age was 64 for the hospitalized patients, 50 years for the outpatient clinic and 47 years at the doctors' office. Cardiovascular diseases were most common. This group of diagnoses was twice as common in hospitalized patients (40.9%) than in the ambulatory group: 19.5% (outpatient clinic) and 15.2% (private practice). In contrast, psychiatric diagnoses were 5 times more frequent in the practice than in the hospital (11.6% vs. 2.4%) and musculoskeletal diseases were even 7 times more common in the practice (13.3% vs. 1.9). 93% of the hospitalized patients had one of the 25 most prevalent diagnoses. This percentage was substantially lower in patients of the outpatient clinic (73%) and of private practice (64%). An additional diagnosis was posed for every other patient in the hospital or at the outpatient clinic, in the practitioners office only for every fourth patient. The analysis shows that hospitalized patients often have a different set of diagnoses than ambulatory ones. To guarantee a broad internistic training outpatient clinics are of great importance. A part of internistic education, however, can also be obtained by cooperation with practitioners. PMID- 10686811 TI - [Primary HIV infection, how to recognize it?]. AB - The primary HIV infection is the period of time following HIV inoculation. Its manifestations are diverse. We present here some clinical cases: a mononucleosis like syndrome with fever, angina, lymphadenopathy and skin rash, a frequent picture, with among other signs, flu-like symptoms, lymphocytic meningitis and facial paresis. In presence of those nonspecific clinical pictures, it is important for the primary health care physician to consider primary HIV infection, detect a history of exposure and order HIV-tests including p24 antigenemia. On one side, an early treatment blocks replication and dissemination of HIV in the body and brings an amelioration of prognosis. On the other side, the patient is particularly infectious during this phase and should take appropriate preventive measures. PMID- 10686812 TI - [Sarcoidosis: acute versus chronic--2 case reports]. AB - Sarcoidosis manifests itself with different symptoms and can thereby follow an acute or chronic course. We will discuss one case each. Case 1 involves a 41-year old patient with Loefgren's syndrome, showing the typical trias of erythema nodosum, bilateral hilar lymphadenopathy and acute polyarthritis. With systemic steroid treatment the symptoms disappeared rapidly. In case 2 we present a 39 year-old-man with chronic sarcoidosis for several years with no vital threat. His organ involvement includes chronic dacryocystitis. Systemic steroids relieved his symptoms and the dacryo-rhinostomy could perhaps have been prevented had the therapy been started earlier. Evaluating the diagnostic and therapeutic procedure can be difficult because there are no standard guidelines. A vital threat demands rapid diagnosis and effective therapy. The chronic illness requires a thorough evaluation of the appropriate therapy. We discuss the criteria for diagnosis, therapy and follow up. PMID- 10686813 TI - [Phantom limb after amputation--overview and new knowledge]. AB - Almost all patients who have an extremity amputated will experience a phantom limb. Amputations of other parts of the body can also cause phantom sensations. One fourth of all women who undergo mastectomy relates phantom breast sensations. Phantoms are common following rectum amputation and may be significant as indicators of rectal tumor relapse. Visual phantoms can appear in patients who undergo eye amputation. Phantom phenomena occur after tooth extraction, ureterocystectomy, penectomy, plexus avulsion or spinal cord injury. The causes underlying phantom sensations are unknown. Sensory deprivation in animals causes reorganization of the cortical and subcortical maps: the areas representing the deprived input shrink and the neighbouring areas expand. The mapping allocates areas to represent the most used peripheral inputs. Every level of the nervous system seems to exhibit plasticity. The primary site seems to be the cortex. The cellular basis of plasticity is unclear. Significant sensory and motor reorganization was found in humans suffering phantom pain. There was a strong relationship between the amount of cortical reorganization and the intensity of phantom pain. These findings may influence the rehabilitation of the amputee. It was shown that pain and cortical reorganization can be reduced or even prevented by the active use of prostheses. PMID- 10686814 TI - [Postoperative nausea and vomiting: what can be done?]. PMID- 10686815 TI - [Erythema nodosum in grade I sarcoidosis]. PMID- 10686816 TI - [Biomechanical significance of the acetabular roof and its reaction to mechanical injury]. AB - INTRODUCTION: The introduction of morphometry into the quantitative analysis of the bone system and functional adaptation of acetabulum to mechanical damages and injuries enabled a relatively simple and acceptable examination of morphological acetabular changes in patients with damaged hip joints. Measurements of the depth and form of acetabulum can be done by radiological methods, computerized tomography and ultrasound (1-9). AIM OF STUDY: The aim of the study was to obtain data on the behaviour of acetabular roof, the so-called "eyebrow", by morphometric analyses during different mechanical injuries. METHOD: Clinical studies of the effect of different loads on acetabular roof were carried out in 741 patients. Radiographic findings of 400 men and 341 women were analysed. The control group was composed of 148 patients with normal hip joints. Average age of the patients was 54.7 years and that of control subjects 52.0 years. Data processing was done for all examined patients. RESULTS: On the basis of our measurements the average size of female "eyebrow" ranged from 24.8 mm to 31.5 mm with standard deviation of 0.93 and in men from 29.4 mm to 40.3 mm with standard deviation of 1.54. The average size in the whole population was 32.1 mm with standard deviation of 15.61. Statistical analyses revealed high correlation coefficients between the age and "eyebrow" size in men (r = 0.124; p < 0.05); it was statically in inverse proportion (Graph 1). However, in female patients the correlation coefficient was statistically significant (r = 0.060; p > 0.05). The examination of the size of collodiaphysial angle and length of "eyebrow" revealed that "eyebrow" length was in inverse proportion to the size of collodiaphysial angle (r = 0.113; p < 0.05). The average "eyebrow" length in relation to the size of collodiaphysial angle ranged from 21.3 mm to 35.2 mm with standard deviation of 1.60. There was no statistically significant correlation between the "eyebrow" size and Wiberg's angle in male (r = 0.049; p > 0.05) and female (r = 0.005; p > 0.05) patients. The "eyebrow" length was proportionally dependent on the size of the shortened extremity in all examined subjects. This dependence was statistically significant both in female (r = 0.208; p < 0.05) and male (r = 0.193; p < 0.05) patients. The study revealed that fossa acetabuli was forward and downward laterally directed. The size, form and cross-section of acetabulum changed during different loads. Dimensions and morphological changes in acetabulum showed some but unimportant changes in comparison to that in the control group. These findings are graphically presented in Figure 5 and numerically in Tables 1 and 2. DISCUSSION: The study of spatial orientation among hip joints revealed that fossa acetabuli was forward and downward laterally directed; this was in accordance with results other authors (1, 7, 9, 15, 18). There was a statistically significant difference in relation to the "eyebrow" size between patients and normal subjects (t = 3.88; p < 0.05). The average difference of "eyebrow" size was 6.892 mm. A larger "eyebrow" was found in patients with normally loaded hip. There was also a significant difference in "eyebrow" size between patients and healthy female subjects (t = 4.605; p < 0.05). A larger "eyebrow" of 8.79 mm was found in female subjects with normally loaded hip. On the basis of our study it can be concluded that the findings related to changes in acetabular roof, the so-called "eyebrow", are important in diagnosis, follow-up and therapy of pathogenetic processes of these disorders. PMID- 10686817 TI - [Long-term patency of reversed and in situ femoro-popliteal bypasses]. AB - INTRODUCTION: The small choice of graft materials is one of the greatest problems in femoro-popliteal (F-P) bypass reconstructions. Besides all biosynthetics(2-5) and synthetics(6) graft materials, there is no right alternative for autologous saphenous vein graft in F-P reconstructions. There are two main techniques for F P reconstructions: "reversed" and "in situ". The aim of this study is the comparison of the long-term patency between "reversed" and "in situ" F-P bypasses. PATIENTS AND METHODS: In the study were included 191 patients with "reversed" and 99 patients with "in situ" F-P bypass grafts operated on between 1988 and 1994. There were 153 (80.10%) male and 38 (19.90%) female patients in the group with "reversed" bypass, and 78 (78.78%) male and 21 (21.22%) female patients in the group with "in situ" bypass. The average age of all patients was 59.04 (27-80) years. Eighty five (44.5%) patients in the group with "reversed" F P bypass had diabetes mellitus and 43 (43.43%) in the group with "in situ" bypass. One hundred and fifty two (79.68%) patients in the group with "reversed" bypass were cigarette smokers and as 80 (80.8%) in the group with "in situ" bypass. In Table 1 the Fontain classification of occlusive diseases in operated patients is presented. The early proximal reconstructions were performed in 49 patients with "reversed" and 16 patients with "in situ" bypasses (Table 2). The associated proximal reconstructions were performed in 21 patients with "reversed" and in 14 patients with "in situ" bypasses (Table 3). All patients were controlled by physical and Doppler ultrasonographic examination immediately after the operation, after 1, 3, 6 months, and then every year postoperativelly. In cases with suspected graft occlusion or any other complication, control angiographic examinations was also performed. The statistical analysis of the results was done using "Life table" analysis. RESULTS: The patients were followed up from 3 to 10 years. The results of "life-table" analysis are presented in Tables 4-8 and Graph 1. The "in situ" technique showed statistically significant better long-term patency compared to "reversed" technique, after 2 and 10 years (p < 0.05). The immediate patency in cases with "reversed" bypass was 98.96%, while limb salvage was 97.91%. In the same group long-term patency was 72.8% and limb salvage 73.9%. In the group with "in situ" bypasses the immediate patency as well as limb salvage were 96.97%. In the same group long-term patency was 73.8% and limb salvage 77.2%. In Table 5 potential advantages of the "in situ" F-P bypass technique are shown (16-21). However, there are controversial data on clinical results of both bypasses. Some authors described better long-term results of the "in situ" F-P bypass technique (28-30), while according to other data there are no significant differences between these two bypass groups (31 33). Most authors emphasized the two advantages of "in situ" bypasses in F-P reconstructions: a small diameter of the saphenous vein; in cases with pure run off (34-36). PMID- 10686818 TI - [Descriptive and epidemiologic characteristics of patients with malignant upper urothelial tumors in the endemic area of Lazarevac]. AB - Although there is permanent increase in incidence of malignant upper urothelial tumours [1, 2], these malignancies are rare neoplasms in relation to both all malignant tumours and urotract tumours. Upper urothelial tumours, i.e. tumours of the renal pelvis and ureter are more frequent in the regions affected by endemic nephropathy [3-5]. The aim of this paper was to describe the main epidemiological characteristics in patients with upper urothelial tumours (UUT) in endemic nephropathic (EN) foci in Lazarevac. We analyzed 73 patients treated at the Institute of Endemic Nephropathy, Lazarevac and the Institute of Urology and Nephrology, Belgrade, from January 1, 1992 to December 31, 1994. The descriptive epidemiological methods was used. The characteristics in patients with histopathologically confirmed upper urothelial tumours were examined. The diagnosis was made on the basis of the clinical picture, echo-sonographic and radioscopic examinations, intravenous and infusion urography and retrograde pielography. With genealogic analysis, a genealogical tree as far as the fourth degree of kinship for each patient, was made both for urothelial tumours and endemic nephropathy. The average age of the patients at the time of diagnosis was 64.2 years, and the majority of the patients (59%) was in the seventh decade of life (Figure 1). Our results are in accordance with the results of other authors who examined the patients with upper urothelial tumours in the regions with endemic nephropathy and out of them [7, 8, 12, 14]. Females were more affected than males (1.4:1). These results are in accordance with the results of other authors who studied the endemic regions [7, 11, 13]. Foreign authors found that males were more affected by upper urothelial tumours [9, 10]. In view of anatomic localization of tumours (Table 2) our results are in accordance with results of the studies carried out in endemic [11, 12, 15, 19] and non-endemic regions [8]. The majority of patients were rural population and lived in villages known as endemic foci (89%) (Table 1). Agriculture was their main or additional occupation. A large number of UUT patients (67%) had endemic nephropathy as well. The other authors from our country found that farmers were most affected [17, 18]. In foreign studies, there are no data on the fact that farming is risk for the appearance of upper urothelial tumours. The family agglomeration of UUT and EN in UUT patients has been observed in all degrees of relation, especially in the second and third generations (Table 3). The obtained results are comparable with hypotheses on a possible mutual or the same aetiological factor for both diseases, which is in accordance with the results of other authors who studied the endemic regions [6, 7, 12, 13]. PMID- 10686819 TI - [Angioblastic lymphadenopathy--its course and prognosis]. AB - INTRODUCTION: In recent years important advances have been made in the understanding of angioimmunoblastic lymphadenopathy since substantial controversy has been related to the name, course, prognosis and therapy of the disease. It was first recognized in the Kil Classification as a low risk T-cell lymphoma [5], and omitted from the most widely used Working Formulation for clinical purposes. According to the criteria of REAL (Revised European American Lymphoma), classification angioimmunoblastic lymphadenopathy (AILD) is one of peripheral postthymic T cell lymphomas that are an immunologically defined category of non Hodgkin's lymphomas originating from the peripheral lymphatic tissues. Morphologically, AILD is characterized by partially or completely obliterated sinuses and frequent infiltration of the pericapsular tissue and substantial proliferation of epithelioid, postcapillary venules. Cytologically, polymorphous cellular infiltration with immunoblasts, transformed lymphoid cells, polyclonal plasma cells, eosinophils and epithelioid cells are found. Clinically, rapid occurrence of systemic symptoms in elderly individuals (sixth and seventh decades of life) with generalized lymphadenopathy, hepatosplenomegaly and cutaneous maculo-papulous or erythematous rash is noted. The patients are characterized with hyperimmune condition in the form of Coombs' positive haemolytic anaemia, polyclonal hypergamma-globulinaemia and liability to infections [8, 9]. In spite of numerous suggestions, therapeutic consensus has not been achieved, and the reported survival ranges from 1 to 30 months [10, 11]. Therefore, this information suggests an aggressive form of the disease with the 60% mortality rate. METHODS: At the Institute of Haematology of the Clinical Centre of Serbia in Belgrade in the last five years, from 1993 through August 1998, nine patients were diagnosed with AILD according to the results of pathohistological examination of the extirpated peripheral lymph nodes and the correlation with clinical picture and relevant laboratory findings. RESULTS: Clinical characteristics of nine patients in whom AILD was diagnosed after lymph node biopsy are given in Table 1. The group consisted of 6 men and 3 women, mean age 53. Eight patients were in advanced stage of the disease at the time of the diagnosis (III and IC CS), while the patient in II CS stage had a large tumorous mass (M+). All patients had initial systemic symptoms. Five of them developed fever with chills. Three patients had evidence of extranodal infiltration of the bone marrow. Infiltration of the liver was suspected in two patients according to aberrant hepatogram values, although pathohistological verification was not obtained. In one patient lung infiltration was histologically verified in addition to bone marrow and liver infiltration. All patients had peripheral lymphadenopathy, and most of them hepatosplenomegaly, as well. Three patients had the so called bulky form of the disease since the diameter of the largest tumour exceeded 10 cm. On admission, most were in poor overall condition, and only two were apparently healthy. Knowing that AILD is basically an immunoregulatory disease and that the described cases of association with systemic diseases of the connective tissue and some drugs were implied in the triggering of AILD, Table 2 shows important information obtained form histories of these patients. Namely, 7 of 9 patients had cutaneous changes suggestive of erythematous or maculopapular rash, while three had received corticosteroid therapy for months before AILD was diagnosed since toxoallergic exanthema had been incorrectly suspected. Three patients received gold sodium thiosulfate therapy for rheumatoid arthritis, while four had history of allergy to drugs and pollen. Table 3 shows laboratory results: anaemia was present in 8 of 9 patients, it was severe in three with haemoglobin values of 67 g/L, 72 g/L and 50 g/L, respectively. Five patients had haemolysis. A PMID- 10686820 TI - [Organization of the program for prevention and control of nosocomial infections in France]. AB - Nosocomial infections (NI) are infections acquired in hospitals. The aim of this paper is to describe the organization of NI control program in France. The organization of this program started in 1988 by the formation of the Infection Control Committees in hospitals. Their role has been to organize the surveillance of NI, to verify basic measures of hygiene, safety of invasive procedures, disinfection and sterilization procedures, occupational safety, and to organize the continued education of health staff members. Operational teams have also been established in hospitals. At the national and regional levels, the National and Regional Infection Control Committees were established in 1992 in order to define the national policy for the treatment of actual infection problems in hospitals and to organize the co-operation between hospitals. In addition, many research projects concerning NI have been conducted and a number of international scientific meetings regarding this subject have taken place in France. According to the two surveys conducted at the national level, in 1990 and 1996, the prevalence rate of NI was found to be 7.4 and 7.6%, respectively. The infection control program could probably be integrated, in the future, into the Hospital Risk Management Program. PMID- 10686821 TI - [Epidemiologic and epizootiologic significance of ticks (Acari: Ixodidae, Argasidae)]. PMID- 10686822 TI - [Experimental models for studies of chronic renal insufficiency]. PMID- 10686823 TI - [Chemotherapy of small-cell carcinoma of the bronchi]. PMID- 10686824 TI - [Electrophysiologic variants in late infantile neuronal-ceroid lipofuscinosis]. PMID- 10686825 TI - [Heterotopic liver tissue in the fundus of the gallbladder]. PMID- 10686826 TI - [Peliosis of the spleen]. PMID- 10686827 TI - [Anaphylactic reaction caused by histamine H2 receptor antagonists in a patient with bronchial asthma]. PMID- 10686829 TI - Hepatitis C--global prevalence (update). PMID- 10686828 TI - Lassa fever, case imported to Germany. PMID- 10686830 TI - Vaccines and biologicals. Report of the Strategic Advisory Group of Experts (SAGE), November 1999. PMID- 10686831 TI - Influenza. PMID- 10686832 TI - Long-term cephalometric changes in untreated adults compared to those treated with orthognathic surgery. AB - A surprisingly large amount of long-term remodeling of facial structures has been noted in the period between 1 and 5 years post-orthognathic surgery. To evaluate whether these changes are greater than in patients with similar morphology who did not have surgery, long-term changes in hard tissue landmarks were examined in 33 untreated adults and compared to long-term changes in skeletal Class II surgery patients who underwent maxillary impaction, mandibular advancement, or both. Although the changes were small in both groups, mean changes were greater in the surgical patients; the surgical patients also showed a higher percentage of significant changes. Horizontal changes were in a forward direction in the untreated group and a backward direction in the surgical groups. We conclude that normal adult growth cannot account for the long-term changes observed following jaw surgery. In some instances, postsurgical changes leading to relapse continue much longer than would have been expected. PMID- 10686833 TI - Psychologic implications of surgical-orthodontic treatment in patients with anterior open bite. AB - Two hundred eighty-two patients who received surgical-orthodontic treatment to correct anterior open bite were retrospectively evaluated by interview and questionnaires to determine the motivation and expectations before treatment, experience during treatment, psychosocial impact, functional and esthetic results, and satisfaction. All patients underwent a Le Fort I osteotomy, and 126 patients also received a bilateral sagittal split advancement osteotomy. The mean follow-up was 6 years. The most important reasons for treatment, as cited by the patients, were biting and chewing problems (28%), dissatisfaction with facial appearance (26%), and symptoms of temporomandibular joint (TMJ) dysfunction (21%). Patients with anterior open bite had a critical attitude toward facial appearance; therefore, esthetic aspects should be taken seriously. The expectations on chewing ability, phonetics, nasal passage, and facial appearance were met by the treatment; however, expectations on TMJ function, interincisal relationship, and biting ability were not completely fulfilled. There was a subjective improvement of TMJ sounds in 27% and a worsening in 14% of the patients. Dysesthesia of the infraorbital nerve was noticed in 4% of patients and of the mental or inferior alveolar nerve in 23% of the patients. Chewing and biting abilities improved in 53% and 73%, respectively. Facial appearance, self confidence, and social interaction had improved. Patients had expected more information before and psychologic support after treatment. Despite the relapse of open bite in 20% of the patients, 75% were satisfied with the dental and 85% with the facial appearance. PMID- 10686834 TI - Psychosocial predictors of high-risk patients undergoing orthognathic surgery. AB - The purpose of this analysis was to identify a set of predictor variables that are prospectively related to postsurgical outcomes. Specifically, psychosocial characteristics were sought to predict postsurgical outcomes. The 5 Revised Symptom Checklist-90 (SCL-90-R) scales, the neuroticism score of the Eysenck Personality Inventory (EPI), the psychosocial domain score from the Sickness Impact Profile (SIP), and 4 scales from the Oral Health Status Questionnaire (OHSQ) were used as the predictors. A total of 31 male and 86 female subjects participated in this multicenter randomized trial, which compared rigid and wire fixation. Data were collected prior to placement of orthodontic appliances, 1 to 2 weeks presurgery, and at 1 week, 8 weeks, 6 months, and 2 years after surgery. Baseline oral health was used as an indicator of postsurgical oral health functioning. A path analytic model of influences on presurgical oral health was estimated (R2 = 0.43). The results suggest that presurgical screening of demographic characteristics (age, sex, and ethnicity), oral health (the OHSQ), quality of life issues (SIP), and personality features (SCL-90-R), accounts for 23% to 39% of the variance in postsurgical oral health outcomes. The path analysis conducted suggests that a patient's age, ethnicity, gender, and elevated scores on the EPI have indirect effects on postsurgical health. As determined by a 2-stage least squares regression model, 3 variables--the patient's presurgical oral health (per the OHSQ), pre- and postsurgical Global Severity Index (GSI) score from the SCL-90-R, and the psychosocial scale score from the SIP--were found to have a statistically significant impact on postsurgical outcomes. Additionally, the GSI, SIP, and OHSQ are reliable measures in predicting oral health outcomes. PMID- 10686835 TI - Computer-animated comparison of self-perception with actual profiles of orthodontic and nonorthodontic subjects. AB - To determine if motivation for adult orthodontic treatment is influenced by self perception, a computer morphing program was developed to animate discrete digitized photographs of facial profiles. It was hypothesized that orthodontic patients are less tolerant of variations in their profiles than nonorthodontic patients. Sixteen orthodontic and 14 nonorthodontic adult patients were presented with animated distortions of 5 features of the lower third of their own profiles. They were asked to identify the zone of acceptability in the changing profile and to indicate the single most pleasing distortion and their perceived and preferred profiles, for comparison with their actual profile. Although orthodontic subjects did not differ from nonorthodontic subjects in the zone of acceptability of their own profiles, they were less tolerant (smaller zone of acceptability) of variation in features of a standard control face. Orthodontic subjects, however, had a larger disparity between the most pleasing and at least one feature of their actual profile than did the nonorthodontic subjects. Orthodontic and nonorthodontic subjects were equally accurate in their ability to identify their own profile features. This unique method of measuring self-perception offers the clinician the advantage of providing a dynamic range rather than a single point of acceptable change to the patient. Moreover, this interactive computer program will enable patients to actively participate in treatment planning decisions by communicating preferences for variations in facial profile distortions. PMID- 10686836 TI - Patients' evaluation of the final result of sagittal split osteotomy: is it influenced by impaired sensitivity of the lower lip and chin? AB - The purpose of this article was to evaluate whether neurosensory disturbances such as impaired sensitivity of the lower lip and chin influence patients' final evaluation of the treatment result. Information about the patients' degree of satisfaction and about lip and chin sensitivity were obtained from final follow up documents of 215 patients. All patients had undergone sagittal split ramus osteotomy for mandibular advancement. In patients with normal sensitivity on both sides of the lower lip and chin, the degree of satisfaction was equally distributed at a very high level through all age groups. Among those with some degree of numbness on either or both sides of the lower lip and chin, patients in the younger quarter and middle half of the group were as satisfied as those with normal sensitivity, while those in the oldest quarter with impaired sensitivity demonstrated a lower degree of satisfaction than the rest of the patients. Although the difference was not statistically significant, the finding strongly indicates that older patients seem to suffer more from neurosensory disturbances than do younger patients with similar conditions. PMID- 10686837 TI - Three-dimensional computer-assisted simulation combining facial skeleton with facial morphology for orthognathic surgery. AB - The purpose of this study was to use a 3-dimensional (3D) computer-aided design (CAD) simulation system to plan surgical procedures and predict postoperative changes in orthognathic surgery patients. A computer-generated imaging model was developed by combining a 3D reconstructed cephalometric skeletal image and a laser-scanned facial surface image. Moreover, postoperative data were studied and linked to the simulator model for programming and executing simulated surgical procedures. Interactive editing capabilities allow surgeons to operate CAD surgical simulation, and predicted results can be presented graphically and numerically. The results indicate that the integration of 3D images and CAD techniques have a potential for simulating surgery and providing graphic information to patients in obtaining an informed consent. PMID- 10686838 TI - Duration of orthodontic treatment involving orthognathic surgery. AB - The purpose of this study was to analyze factors influencing the duration of treatment in a sample of patients treated by a combined orthodontic/orthognathic surgery approach. Presurgical and postsurgical treatment times were assessed for 315 patients processed through an orthognathic team over a 7-year period. The median total treatment duration for all patients was 21.9 months, the median presurgical duration was 15.4 months, and the median postsurgical duration was 5.9 months. Treatment involving extractions resulted in significantly increased presurgical and total treatment times. Treatments performed in the university clinic showed reduced presurgical duration and increased postsurgical duration compared to treatments carried out by specialists outside the university. Presurgical, postsurgical, and total treatment times were significantly reduced when the orthodontist had treated 10 or more patients during the period. An earlier regimen of orthodontic treatment did not appear to have a significant effect on treatment duration. PMID- 10686839 TI - Dental arch size in healthy human permanent dentitions: ethnic differences as assessed by discriminant analysis. AB - Race and ethnicity variably influence the form of the human craniofacial complex. In the present study, the effects of ethnicity and sex on the global size of normal adult dental arches were analyzed. The dental arches of 47 northern Chilean mestizos (25 men, 22 women) and 95 northern Italian Caucasians (50 men, 45 women) were cast in stone. All subjects had a complete dentition in both arches. In all models the coordinates of dental cusp tips were digitized using an image analyzer. The center of gravity of each tooth was computed and arches were interpolated using a polynomial model (y = ax + bx2 + cx3 + dx4). In all arches, the intercanine, intermolar, and mid-intercanine to mid-intermolar distances were computed from the dental centers of gravity. These arch distances were entered in a linear discriminant function analysis. The polynomial model accurately interpolated data points in all instances, and most of the dental arch form was determined by the first and second degree coefficients. On average, Italian Caucasian arches were smaller than Chilean mestizo arches. Male mean distances were larger than female distances regardless of ethnic group or arch. The linear discriminant analysis performed between male and female arches within ethnic groups was significant only for both Italian Caucasian arches, but the percentage errors for the classification of a new individual were very high (about 30%). Conversely, Italian Caucasian arches could always be discriminated from Chilean mestizo arches of the same sex with a much smaller error. PMID- 10686840 TI - An unusual treatment with sagittal split osteotomy: report of a case involving an odontoma. AB - Sagittal split osteotomy is one of the most commonly performed surgical techniques in the world and has been modified by many authors. The efficacy of this operation has been studied by many groups. When performing this surgery, there should be adequate contact of wide, cancellous bone surfaces, which guarantees excellent and rapid bony union in the desired position. In the present article, treatment of mandibular prognathism with open bite by sagittal split osteotomy with an odontoma in the third molar area is presented. PMID- 10686841 TI - Geometric considerations when planning an asymmetric genioplasty. AB - The sliding osteotomy of the inferior border of the mandible, otherwise known as genioplasty, has often been described in the world literature with regard to diagnosis and treatment planning. However, the treatment of the asymmetric chin has received little attention. Moreover, diagnosis and treatment planning of asymmetric chins with concomitant orthognathic surgery is completely lacking from the literature. The complexity of surgically correcting asymmetric chins, compounded with complex, bimaxillary orthognathic surgery, is an extremely challenging task. This article looks at geometric considerations when planning the surgical correction of an asymmetric chin following a protocol of data collection, model surgery, diagnosis, and treatment planning. Clinical experience in the form of a case presentation will demonstrate the millimetric precision that can be achieved when planning corrective genioplasty in an asymmetric patient undergoing concomitant orthognathic surgery. PMID- 10686842 TI - Vertical chin augmentation with interpositional porous polyethylene implants: a histologic study in monkeys. AB - The objective of this paper was to evaluate histologically the tissue reaction in the chin after a vertical augmentation using interpositional porous polyethylene (PPE) implants in monkeys. Six monkeys (Cebus apella) underwent an anterior horizontal mandibular osteotomy with implantation of an interpositional PPE implant to increase the vertical height. The animals were sacrificed 5 months postoperatively. Histologic preparations were stained with hematoxylin and eosin. The perimeter of the interface between the implant and the bone, the implant and the trabecular space, and the implant and the fibrous capsule were quantified using the NIH Image Analysis System (Image 1.60/PPC). In addition, the Tukey test was done. The study demonstrated that bone growth takes place within the pores of the implant; a fibrous capsule exists in some animals, where the implant has contact with the periosteum and mentalis muscle with few chronic inflammatory cells; and the 3 different tissues responded in statistically different manners. Perimeter analysis revealed 68.9% implant-bone contact, 22.9% implant-fibrous tissue contact, and 8.2% implant-trabecular space contact. PMID- 10686843 TI - Demineralized intramembranous bone matrix augments the healing of endochondral bone graft. AB - The aim of this study was to examine the osteogenic potential of demineralized bone matrix prepared from intramembranous bone (DBMIM) and to examine its effects on the healing of endochondral autogenous bone grafts. Twenty-four defects in 24 New Zealand white rabbits were used as experimental groups. Twelve defects were grafted with endochondral bone, and the other 12 defects were grafted with endochondral bone with DBMIM (EC-DBMIM). One rabbit from each group was sacrificed on days 1, 2, 3, 4, 5, 6, and 7 postgrafting, and the remaining 5 rabbits from each group were sacrificed on day 14 postgrafting. Another 8 defects in 4 rabbits were used as control groups: 4 defects were left empty (passive control), and 4 defects were grafted with rabbit skin collagen (positive control). They were all sacrificed at 14 days after grafting. Serial sections were made across the whole defect. Quantitative analysis was performed on 100 sections of the 14-day experimental groups by image analysis. Four hundred fourteen percent more new bone was formed in defects grafted with composite EC DBMIM than in those grafted with endochondral bone alone (P < 0.0001). No bone was formed in either passive or positive controls. Histologic examination of the bone grafts revealed intermediate-stage cartilage, and immunohistochemical examination revealed earlier vascularization in the composite EC-DBMIM groups. In conclusion, DBMIM has extremely high osteoinductive properties and greatly enhances the integration of endochondral bone with defects of intramembranous bone in origin. PMID- 10686844 TI - Soft tissue changes associated with incisor decompensation prior to orthognathic surgery. AB - A retrospective cephalometric study employing angular and linear measurements was undertaken to examine soft tissue changes associated with incisor decompensation prior to orthognathic surgery. Subjects were divided into 3 groups based on the presenting malocclusion, and radiographs for each subject were traced and subsequently digitized. The results showed that incisor decompensation was achieved more markedly in Class II division 2 and Class III patients. In Class II division 1 subjects, changes in the mandibular incisor inclination were contrary to what was anticipated. The soft tissue change/dental change ratio following incisor decompensation showed that the soft tissues were least affected in the Class II division 2 group. This may be due to the increased tone of the soft tissues in this group, which resisted the effects of the incisor change. Alternatively, it may indicate that the soft tissue "drape" is not in close approximation to the dentition and therefore dental changes are not transmitted to the soft tissues. PMID- 10686845 TI - Stability of Le Fort I osteotomy in maxillary advancement: review of the literature. AB - Stability of the skeletal segments repositioned during orthognathic surgery is still a concern in maxillofacial surgery. In an attempt to establish a consensus about one of the most frequently performed repositioning surgeries, the literature from 1985 to 1999 concerning stability of Le Fort I osteotomy in maxillary advancement was reviewed. There have been many problems in interpreting the results of the analysis because of differences in the design of the studies and the multifactorial nature of the disorder. For this reason each problem that emerged in the literature is analyzed and discussed. PMID- 10686846 TI - Obstructive sleep apnea: a principal component analysis. AB - A principal component analysis was performed on the cephalometric variables of 100 male obstructive sleep apnea (OSA) patients. Thirty cephalometric variables of cervicocraniofacial skeletal morphology were reduced to 8 principal components (PCs), which described 83.2% of the total variance. Sixteen cephalometric variables of hyoid bone position and head posture were reduced to 4 PCs, which described 85.5% of the total variance. Twenty cephalometric variables of upper airway soft tissue were reduced to 7 PCs, which described 83.7% of the total variance. These PCs described the actual characteristics of the OSA patients examined. For further analysis of PCs, stepwise multiple regression analysis was chosen. Two dependent variables of interest are the minimal distance of posterior pharyngeal airway space (PASmin) and the Apnea-Hypopnea Index (AHI). Seven PCs accounted for 79.4% of the variance of PASmin and 3 PCs accounted for 37.6% of the variance of AHI. Both principal component analysis and multiple regression analysis provide multivariate data analysis that is very useful in sorting out and clarifying the complexity of the interrelated cervicocraniofacial skeletal morphology and upper airway soft tissue in OSA patients. PMID- 10686847 TI - Morphometry of the mandible in prepubertal craniofacial microsomia patients following an inverted L osteotomy. AB - The aim of this study was to undertake finite element modeling of craniofacial microsomia (CFM) patients who exhibited a unilateral mandibular deformity that was surgically corrected by an inverted L osteotomy and autogenous bone graft. Preoperative, 1-year, and 3-year postoperative anteroposterior cephalographs of 14 consecutive children (mean age, about 9 years) with CFM were employed. All cephalographs were scanned, and 8 homologous mandibular landmarks were digitized in triplicate (digitization error < 1%; P > 0.05). Average mandibular geometries, scaled to an equivalent size, were generated using Procrustes superimposition and subjected to analysis of variance. Results showed that while the mean pre- and early postoperative mandibular configurations marginally failed to differ statistically, finite element modeling of the affected mandibular ramus showed a positive allometry (a lengthening by about 10% to 12%) and marked anisotropy, presumably reflecting the surgical procedure carried out in that region. For the preoperative and late postoperative means, the configurations were statistically different (P < 0.002), showing a lengthening of the ramus (about 14%) while the unoperated side had a high degree of isotropy. The early and late postoperative configurations were also statistically different (P < 0.002). The operated ramus showed an increase in length (about 15%), but the corpus showed a decrease in size (about 10%) and marked anisotropy that reflected remodeling associated with the surgical procedure. Although mandibular morphology is improved in CFM patients who are surgically treated by an inverted L osteotomy, some relapse is evident due to deficient growth, localized in the mandibular ramus and body of the affected side. PMID- 10686848 TI - Use of articulators in the United Kingdom by consultant orthodontists in planning orthognathic surgery. AB - This questionnaire survey aimed to investigate articulator use in orthognathic surgical planning by consultant specialists in the United Kingdom (UK). A total of 205 questionnaires was sent to all consultant orthodontists in the UK. One hundred thirty questionnaires were completed, representing a 63.4% response rate. Consultants had been in their post for a median of 10.5 years (range 0 to 30 years) with a mean of 11.3 (SD 8.4) surgical cases per year. When asked which cases had been planned on a semiadjustable articulator, two thirds of consultants (67.7%) planned maxillary single-jaw procedures, 45.4% planned mandibular single jaw procedures, and 77.7% planned bimaxillary procedures. While 98.5% of consultants reported access to at least one type of articulator, 14.6% of maxillary single-jaw and 5.4% of bimaxillary surgery was not planned on any articulator. Twice as many consultants who had been in their post fewer than 11 years had been trained on a semiadjustable articulator, compared to those who had more than 11 years of experience. Semiadjustable articulators were the most popular for planning orthognathic surgery. Consultants with less than 11 years of experience completed more surgical cases each year and were more likely to have been trained on a semiadjustable articulator than consultants with more than 11 years of experience. However, no link exists between the age of the consultant and the type of articulator selected for planning; this suggests that more mature consultants have received further training on contemporary articulator systems since receiving their accreditation. PMID- 10686849 TI - Condylar resorption 2 years following active orthodontic treatment: a case report. AB - We recently treated a patient with degenerative disease of the temporomandibular joint. A healthy, 12-year-old female with bilateral high maxillary canines presented for orthodontic treatment. Two years after active orthodontic treatment, at age 17, symptoms in her temporomandibular joint manifested and progressed. By the time she revisited our hospital at age 21, the patient had developed an anterior open bite with a long, slender facial appearance. Cephalometric analysis showed shortening of the ramus and backward and downward rotation of the mandible. Imaging studies revealed severe deformity and resorption of the bilateral condyles. Her occlusal and morphologic changes seemed to be caused by degenerative disease of the temporomandibular joint. PMID- 10686850 TI - Dental applications of amorphous calcium phosphates. AB - Certain commercial materials and equipment are identified in this paper to specify the experimental procedure. In no instance does such identification imply recognition or endorsement by the National Institute of Standards and Technology or the ADA Health Foundation, or that the material or equipment identified is necessarily the best available for the purpose. PMID- 10686851 TI - The origins of Enamelon remineralizing fluoride toothpaste. PMID- 10686852 TI - Laboratory enamel solubility reduction and fluoride uptake from enamelon dentifrice. AB - The presence of calcium-releasing ingredients in toothpastes containing sodium fluoride is usually avoided to prevent loss of active fluoride due to the formation of the insoluble calcium salt. The purpose of this study was to determine the bio-availability of fluoride from Enamelon Toothpaste (E), which simultaneously supplies fluoride, calcium and phosphate salts from a dual dispensing package. Fluoride uptake into artificially lesioned enamel cores due to the use of the test dentifrice (E) diluted 1:3 in whole human saliva was compared to that from a clinically proven effective sodium fluoride/silica USP reference toothpaste (C) and a non-fluoride control (E-w/o F). Enamel solubility reduction due to the use of E, C and E-w/o F was measured by determining the quantity of phosphate released to lactic acid buffer before and after treatment of the crowns of molars with 1:3 slurries of the dentifrices in water. Fluoride uptakes and enamel solubility reductions were 5031 +/- 158 ppm and 21.6 +/- 2.2% for E, 1915 +/- 39 ppm and 13.6 +/- 2.0% for C, and -3 +/- 2 ppm and 0.8 +/- 1.7% for E-w/o F. The fluoride uptake and enamel solubility reductions from E were significantly greater than from C (p < 0.001, Fisher LSD), and both fluoride containing dentifrices significantly outperformed E-w/o F (p < 0.001). The laboratory results indicate that the calcium and phosphate salts delivered by the remineralizing Enamelon dentifrice increase the bioavailability of fluoride to substantially exceed that of the clinically proven standard dentifrice. PMID- 10686853 TI - Remineralization by fluoride enhanced with calcium and phosphate ingredients. AB - The effectiveness of fluoride ions provided by toothpastes and mouthrinses in promoting remineralization can be limited by the low concentrations of calcium and phosphate ions in saliva. The purpose of this study was to determine whether improved remineralization can be obtained from toothpastes or mouthrinses that simultaneously deliver fluoride, calcium, and phosphate ions from dual-dispensing systems. Enamel specimens with artificial lesions between 60 and 90 microns deep were cycled 15 times through demineralization for 30 minutes, treated for 5 minutes with an experimental or control fluoride toothpaste or mouthrinse, and remineralized for 60 minutes. In the toothpaste study, surface hardness increased by 11.5 +/- 9.2 and 2.7 +/- 3.6 Vickers hardness units, and enamel fluoride content was 5984 +/- 521 ppm and 3971 +/- 531 ppm for the experimental and control fluoride toothpastes, respectively. Remineralization was confirmed by x ray microradiography. In the mouthrinse study, surface hardness increased by 8.8 +/- 7.7 and 2.2 +/- 3.7 Vickers hardness units, and enamel fluoride content was 6111 +/- 1078 ppm and 3160 +/- 364 ppm for the experimental and control fluoride mouthrinses, respectively. Use of a non-fluoride control mouthrinse led to a decrease in surface hardness of 3.7 +/- 5.2 Vickers hardness units despite a fluoride content of 402 ppm. The results demonstrate that calcium and phosphate supplementation in a toothpaste or mouthrinse can improve remineralization and increase fluoride uptake. PMID- 10686854 TI - Strengthening of tooth enamel by a remineralizing toothpaste after exposure to an acidic soft drink. AB - The effect of remineralizing and conventional toothpaste treatments on the hardness of intact and acid soft drink-etched enamel were assessed in a laboratory study. The remineralizing toothpaste (Enamelon Toothpaste) used in the study contains NaF, and simultaneously provides dissolved calcium, phosphate and fluoride ions. The conventional toothpaste contains NaF in a silica base. Sound extracted maxillary human incisors, mounted in epoxy resin with the facial incisal two-thirds exposed, were polished with 0.3 mu alumina. Groups of ten teeth were exposed to either twenty 5-minute treatments with an acid soft drink (pH 2.4), remineralizing or conventional fluoride toothpaste, or to twenty alternating cycles of a 5-minute protective treatment with either the remineralizing or conventional toothpastes, followed by 5-minute exposures to the acid soft drink, or to 20 five-minute exposures to the acid soft drink followed by 20 five-minute restorative treatments with the remineralizing or conventional toothpastes. Knoop Hardness measurements were made before and after treatment using a 500 g load and 15-second dwell time. Acid soft drink exposure produced a 15.4% drop in hardness compared with 4.9% and 1.6% hardness increases due to treatments of intact enamel with the remineralizing and conventional toothpastes, respectively. Protective treatments using the remineralizing and conventional toothpastes significantly reduced the drop in hardness due to acid soft drink exposure to 3.3% and 6.2%, respectively. Restorative treatments by the remineralizing and conventional toothpastes significantly increased the hardness of the acid soft drink-weakened enamel by 12.1% and 7.3%, respectively. Both toothpastes were effective in inhibiting damage due to acid soft drink exposure, but the remineralizing toothpaste was more effective in hardening intact and decalcified enamel than the conventional toothpaste (p < 0.05). PMID- 10686855 TI - The in vitro demineralization potential of a sodium fluoride, calcium and phosphate ion-containing dentifrice under various experimental conditions. AB - While fluoride has had a dramatic effect in reducing the rate of caries, it has failed to provide complete protection against caries development, and tooth decay is still widely prevalent. The objective of this study was to determine the demineralization prevention potential of a remineralizing dentifrice with fluoride (Enamelon), and an assured supply of calcium and phosphate ions on bovine enamel in a cyclic remin/demin regimen using various experimental conditions. A conventional fluoride toothpaste, Crest, and a placebo dentifrice were used as controls. One-hundred and forty-four ground and polished 4 mm bovine enamel cores were prepared and assigned to various test treatments using six specimens per treatment group. They were subjected to six cyclic treatments consisting of one-minute exposures at room temperature to the test product diluted 1:2 in water, remineralization for a specified time at 37 degrees C, and demineralization for a specified time at 37 degrees C. The enamel specimens were tested for micro-hardness initially, and after the first, third and sixth treatment, remineralization and demineralization cycles using a Buehler Micro hardness Tester with a 200 g load. Differences in micro-hardness between the test groups were analyzed for significance by performing multiple pair-wise comparisons using Bonferroni significance levels at the 95% confidence level. The results of this study showed that Enamelon Toothpaste was generally more effective in preventing demineralization of enamel than the fluoride dentifrice at 0.5 and 1.5 hours, and the non-fluoride control dentifrice at all time measurements. Prevention of enamel demineralization by the dentifrices was affected by changes in demineralization time and pH, but not by changes in remineralization time. PMID- 10686856 TI - Cariostatic effect of a two-part fluoride dentifrice in rats. AB - This study compared the cariostatic effect of a two-part, fluoride-calcium phosphate-containing prototype dentifrice (containing 1100 ppm F) to a conventional, clinically proven effective "gold standard" (1100 ppm F, USP Standard) and an Enamelon placebo dentifrice (0 ppm F) using a rat model. Three groups of 18 Crl:CDR(SD)BR rats were inoculated with S. sobrinus 27352 and fed MIT-200 for 35 days in a programmed feeder. Double-deionized water (DDW) was given ad libitum. The rats were treated twice daily with one two-part dentifrice. Just prior to treatment, each of the two parts of dentifrice were mixed 1:1 and applied undiluted. The rats were euthanized and their mandibular molars scored for caries using the Keyes quantification method. Shrestha SNP caries scores were then calculated. Non-parametric comparisons were done among treatments at the 0.05 level of significance using analyses based on ranks. The mean Shrestha SNP Caries Scores +/- standard deviations for the prototype dentifrice, USP standard and Enamelon placebo were 84.9 +/- 62.8, 101.3 +/- 66.3, and 181.2 +/- 100.1, respectively. Scores for the 1100 ppm F prototype treatment were significantly lower than the USP Standard and the Enamelon placebo. These results indicate that the new prototype dentifrice, containing fluoride-calcium-phosphate, increased the anti-caries efficacy in a rat model when compared with a fluoride-containing USP Standard dentifrice. PMID- 10686857 TI - Caries inhibition in rats by a remineralizing toothpaste. AB - We tested the anti-caries properties of a prototype remineralizing toothpaste containing sodium fluoride, soluble phosphate and soluble calcium, similar but not identical to Enamelon Toothpaste in specific pathogen-free Osborne-Mendel rats. A fluoride-free placebo and sodium fluoride-containing Crest Regular were used as negative and positive control toothpastes, respectively. Sixty weanling rats were randomly distributed into groups of 20, all were inoculated with S. mutans 10449S, ate diet 2000, and drank demineralized water. Each toothpaste, contained in coded tubes, was applied to the rats' teeth, once for one minute/day, five days/week. There was no difference in bacterial recoveries from tooth swabs among the groups at either the 22- or 37-day post-inoculation recovery date. After rat sacrifice and defleshing, heads were randomly coded and scored blindly for carious lesions according to Keyes/Larson methods. Only after scoring was completed were the random codes broken and the treatment groups identified. Both the Enamelon-like product and Crest Regular treatments resulted in lower total enamel caries scores than the placebo (p < 0.001). The sum of smooth surface scores was also lower for the fluoride-containing toothpastes than for the placebo (p < 0.001). However, the Enamelon-like product had greater caries inhibitory properties than Crest Regular on sulcal and approximal tooth surface categories (p = 0.004 and p = 0.045, respectively). Therefore, the Enamelon-like product had superior caries inhibiting properties compared to Crest Regular at these tooth areas. PMID- 10686858 TI - Model for assessment of carious lesion remineralization, and remineralization by a novel toothpaste. AB - This study is of a novel toothpaste which induced remineralization of carious lesions in specific pathogen-free Osborne-Mendel rats. Randomly distributed weanlings in 7 groups (n = 10) each were initially induced to develop carious lesions as a result of a severe dietary and bacteriological challenge. Thus, all were inoculated with S. mutans 10449S, ate diet 2000CS (containing 56% cornstarch) ad libitum, and, upon weaning and for 10 additional days, drank demineralized water supplemented to 10% (w/v) with sucrose (SW) ad libitum. During these 10 days, 6 of the 7 groups of rats were topically treated with a slurried F(-)-free placebo toothpaste. On the 10th day after initial inoculation, two groups were sacrificed, one which had been treated with the placebo and the one which had been untreated. The remaining groups were then provided unsupplemented demineralized water (DW), fed diet 2000CS, and topically treated with either of 5 coded toothpastes: a slurry of the F(-)-free placebo, a similar slurry of a conventional sodium fluoride toothpaste (Crest Regular) containing about 383 ppm F-, or one of three prototype toothpastes formulated by Enamelon, Inc. containing soluble calcium and phosphate salts. Two of these contained 377 ppm F- after mixing their components, either as sodium monofluorophosphate (E MFP) or as NaF (E-NaF). The third prototype contained 0 ppm F- (E-w/o F). Test items were applied to the dentition for one minute/day, five days/week. These 5 remaining groups were sacrificed 13 days after the DW provision. After defleshing, heads were coded randomly and scored blindly for carious lesions. The exchange of DW for SW was associated with highly significant reductions of 10449S recoveries (p < 0.001), but there were no differences in recoveries among the groups as a function of toothpaste type. Total mandibular and maxillary sulcal and smooth surface caries scores were statistically significantly lower for the E NaF-treated group at 23 days than for the 23-day placebo-, E-w/o F(-)-, and conventional NaF toothpaste-treated group. Reductions were most impressive quantitatively on approximal tooth surfaces, where both the E-MFP and E-NaF groups had the lowest scores, but were also statistically significant at sulcal surfaces for the E-NaF group. Thus, this study model can be used to detect significant remineralization effects, as occurred with the prototype toothpaste E NaF. PMID- 10686859 TI - An in vitro assessment and a pilot clinical study of electrical resistance of demineralized enamel. AB - Electrical resistance measurement was evaluated in vitro and in vivo as a method for comparing the remineralizing performance of toothpastes. In the in vitro study, areas of sound enamel on 12 unrestored, mature, extracted human molars and bicuspids, with electrical resistance readings greater than 99.99 M omega, were demineralized to an electrical resistance of 1 to 4 M omega. The teeth were divided into three groups. The change in electrical resistance was measured following a 15 cycle regimen of treatment, demineralization and salivary soaking. Treatments were five-minute exposures to either a 1:2 slurry in saliva of Enamelon (E), a remineralizing fluoride-toothpaste also containing soluble calcium and phosphate ions, a 1:2 slurry in saliva of Crest (C), a conventional fluoride toothpaste (P), or to saliva alone. Demineralization was performed with a 30-minute exposure to 0.1 M lactic acid 50% saturated with calcium hydroxyapatite. The salivary soaking was 1 hour in duration. The mean electrical resistance of the E, C and the saliva treated sites was 63.9 +/- 4.3, 37.6 +/- 9.5 and 2.1 +/- 0.7 M omega, respectively. The final resistance was statistically different for each group (p < 0.05). A pilot clinical study was then conducted to assess the electrical resistance technology in vivo. Eighteen adult subjects with at least one site of early enamel caries with an electrical resistance between 1.0 M omega and 20.00 M omega were randomly assigned to either Enamelon, Crest, or a non-fluoride placebo toothpaste and asked to brush twice daily. After three months, the mean resistance of the test sites was 23.57 M omega for E, 9.61 M omega for C and 13.24 M omega for P. However, the mean resistance changes did not proceed consistently over time. At the end of the study, the electrical resistance measurements on four sites out of twelve in Group E were suggestive of remineralization, whereas measurements on one site out of ten were suggestive of remineralization in Group C and two or three sites out of twelve were suggestive of remineralization in Group P. Progression of demineralization was possibly indicated in only one site, which was in Group C. There were insufficient subjects for statistical significance in the pilot clinical study. There were apparent problems with the variability of some measurements between visits in the in vivo study. Overall, however, the results of both studies indicate that with modifications to the equipment, electrical resistance measurements may be a means of comparing the remineralization performance of toothpastes. PMID- 10686860 TI - Treatment of orthodontic white spot lesions with a remineralizing dentifrice applied by toothbrushing or mouth trays. AB - This pilot study investigated the effect of a remineralizing fluoride dentifrice (Enamelon) on newly formed incipient carious lesions using two methods of application. Teenage orthodontic patients with obvious white spot lesions on their teeth were selected to begin treatment within 30 days after debanding. At baseline, the surface enamel of the test teeth was cleaned by air etching with 50 microns alumina at 150 psi. Lesion size (mm2) was measured using a calibrated periodontal probe and surface appearance was quantified as shiny (0), dull (1), or chalky (2). Twice daily for 3 months, one group with a total of 27 lesions brushed with the remineralizing dentifrice, while a second group with 41 total lesions brushed and used a mouth tray to apply the paste directly to the lesions for 5 min. In order to promote treatment compliance, test group assignment was based on subject and parental preference. After 1, 2, and 3 months, lesion size was reduced by 5% (ns), 10% (p < 0.05) and 22% (p < 0.01), respectively, for the brushing group, and 16% (p < 0.05), 37% (p < 0.01) and 30% (p < 0.01) for the combined brushing and tray group, respectively. The dull appearance of the lesions treated by brushing improved slightly over 3 months. However, the lesions receiving combined brushing and tray applications became significantly (p < 0.01) less dull by 28%, 44% and 61% after 1, 2, and 3 months, respectively, indicating the formation of a shiny, intact surface layer. In conclusion, brushing with a remineralizing dentifrice significantly reduced the size of new orthodontic white spots within 2 months, while brushing combined with topical tray applications reduced lesion size within 1 month with concomitant formation of a shiny enamel surface layer. Thus, the combined brushing and mouth tray treatment significantly accelerated the remineralization process. PMID- 10686861 TI - Clinical evaluation of the effect of a remineralizing toothpaste on dentinal sensitivity. AB - Dentinal hypersensitivity is a common dental problem without a satisfactory solution. Most therapies have focused on either trying to block the stimulation of dentinal nerves or on sealing open dentinal tubules. This study evaluated, in a double blind clinical trial, the effect of Enamelon, a dentifrice containing sodium fluoride, calcium salts and phosphate salts (calcium separated from the phosphate and fluoride by a plastic divider in the tube to prevent interaction) on dentinal hypersensitivity over an 8-week period. A conventional OTC dentifrice containing NaF without calcium and phosphate enhancement served as the control. A third dentifrice, containing sodium monofluorophosphate enhanced by calcium and phosphate salts, was also tested. Based upon subjective anecdotal information, net numbers of sensitive teeth which became non-sensitive, and a logit analysis of the change in the proportion of sensitive teeth over the study time period, Enamelon was the most effective product, and the OTC control the least effective. PMID- 10686862 TI - Evaluation of blood pressure in children and adolescents: a review. AB - Hypertension in children has received minimal attention in the dental literature; this may be the result of comparatively low interest in the subject by the medical community. It is now believed that the processes causing cardiovascular disease morbidity and mortality begin early in life. The implication for prevention is that screening and intervention should commence in childhood. Although the prevalence is lower than in adults, elevated blood pressure during childhood is not uncommon. Dentists may be uniquely positioned to screen children for hypertension. Children from poor families may not use medical services except in emergencies, but may visit a dentist as part of Head Start or through other state or federal programs. By incorporating blood pressure screening for children as well as for adults and with appropriate referral, dentists may contribute to the reduction of cardiovascular disease. This review presents a discussion of blood pressure in children and adolescents, the epidemiology and etiology of elevated blood pressure, the problems associated with hypertension, and recommended procedures for measuring blood pressure in children and young adult patients. PMID- 10686863 TI - Clinical and ultrastructural study of natal and neonatal teeth. AB - The present study was undertaken to evaluate the surface topography of mandibular natal and neonatal incisors at the ultrastructural level using the scanning electron microscope (SEM). The enamel of the teeth exhibited hypoplastic, depressed areas and the incisal edge of natal tooth lacked enamel. In addition, root formation of the teeth was not completed, which correlated with findings that teeth may erupt without root formation. PMID- 10686864 TI - Severe hypodontia: diversities in manifestations. AB - The material comprises 33 cases, 12 boys and 21 girls with 4 or more lacking teeth in the permanent dentition, randomly collected among patients referred to the Department of Pedodontics, University of Bergen. The total number of lacking teeth were 332, mean number in boys was 11.4, in girls 9.3, ranging from 4 to 24 (third molars excluded). The most prevalently lacking teeth were second premolars in both jaws, maxillary lateral incisors, mandibular central incisors and maxillary first premolars. The maxillary central incisors were the most stable teeth, lacking in only one patient. The female group was closest to this "classical" scheme by lacking teeth mostly in posterior segments. In males the anterior segments were most often afflicted. There was no significant difference between right and left sides in both sexes, but in girls the maxillary jaw was more afflicted than the mandibular jaw. The individual analyses of cases showed great diversities in the manifestation of hypodontia. Eighteen of them behaved fairly "balanced" with respect to lack of teeth in the different quadrants. Six cases were lacking most of the teeth in the maxillary jaw (total 34 maxillary teeth versus 11 mandibular). Five cases were lacking most of the teeth in the mandibular jaw (total 30 mandibular teeth versus 14 maxillary). One patient was lacking 10 of his 12 front teeth, but only 4 of his 20 posterior teeth, and one patient was lacking 12 of his 16 posterior teeth, but none of his front teeth. The author cannot offer any explanation for these strange and varying patterns of manifestations. PMID- 10686865 TI - Gingivitis in children with malnutrition. AB - The purpose of the current study was to compare the prevalence and severity of gingivitis and plaque among 291 well- and malnourished children between the ages of 4 to 7 years. Using the National Center for Health Statistics criteria, the children were identified as normal (well nourished), having a height-of-age > 95% of standard. Selected primary teeth of each child were assessed using the Plaque Index (PlI) and the Modified Gingival Index (MGI). The results demonstrated a 100% prevalence of plaque and gingivitis. Also, there was no significant difference in the PlI and MGI in the well-nourished and malnourished groups, nor between males and females. However, there was a trend in the well-nourished group to less plaque and gingivitis when broken down into different stature-by-age percentiles. PMID- 10686866 TI - Orthodontic treatment of a patient with multiple supernumerary teeth and mental retardation. AB - Supernumerary teeth may lead to impaction or ectopic eruption of maxillary incisors, crowding, oronasal fistula, follicular cyst, migration of adjacent teeth and root resorption. In this presentation, an 11-year-old male patient with 4 supernumerary teeth in the maxillary anterior region and a slight mental retardation problem will be presented. After extraction of the supernumerary teeth, the large space created by distal migration of central incisors was closed by orthodontic treatment. Although initial patient compliance was weak and caused difficulties in the course of treatment, orthodontic treatment was completed successfully. PMID- 10686867 TI - Newer Class I cavity preparation for permanent teeth using air abrasion and composite restoration. AB - This study attempts to determine a more effective cavity preparation design, material selection and preparation technique for reducing microleakage in posterior Class I esthetic restorations. An in vitro study using four different cavity designs, for Class I restorations on permanent molars, prepared with two different methods, and restored with three different restorative materials (hybrid composites) was done to evaluate marginal microleakage, and voids occurrence. Two hundred and forty extracted permanent molars were chosen and evaluated for caries, visually, with a dental explorer, and with caries detector dye. The teeth were then randomly divided in two groups (n = 120). In the first group, Class I cavity preparations were performed with air-abrasion. In the second group Class I cavity preparations were performed with #330 bur. The results revealed that cavity preparations prepared with air-abrasion with or without chamfer, and for cavity preparations done with a #330 bur with chamfer and restored with Tetric Flow, had zero microleakage. Cavity preparations done with air-abrasion, without chamfer, and for cavity preparations prepared with #330 bur with chamfer and restored with Tetric Ceram, had zero microleakage score. Cavity preparations done with air-abrasion with chamfer and restored with Herculite had one tooth out of twenty with microleakage, and for cavities without chamfer two teeth had microleakage. Cavity preparations prepared with a #330 bur, without chamfer, and restored with Herculite XRV had four teeth out of twenty with microleakage, and with a chamfer, two teeth had microleakage. These differences were not statistically significant. When comparing Tetric Flow versus Herculite XRV for void formation in cavity preparations prepared with Air abrasion and a chamfer, Tetric Flow had significantly less voids, p < 0.001. When comparing Tetric Ceram versus Herculite XRV for cavity preparations prepared with Air-abrasion and a chamfer, Tetric Ceram had significantly less void formation, p < 0.01 > 0.001. When comparing Tetric Flow versus Herculite XRV for cavity preparations prepared with #330 bur and without a chamfer, Tetric Flow had significantly less void formation, p < 0.02 > 0.01. When comparing Tetric Flow versus Herculite XRV for cavity preparations prepared with #330 bur and a chamfer, Tetric Flow had significantly less void formation, p < 0.001 > 0.001. Caries detection results revealed that the caries detector dye method had significantly higher caries detection scores than explorer p < 001, and than visual inspection p < 0.001. Also inspection with explorer had significantly higher scores than visual inspection p < 0.001. PMID- 10686868 TI - Four types of topical anaesthetic agents: evaluation of clinical effectiveness. AB - In this study, four anesthetic agents of different forms and contents, namely: EMLA 5% Cream (lidocaine 2.5 percent, prilocaine 2.5 percent), Vision-Gel (benzocaine 20 percent), Anesthetic Tabs (tetracaine hydrochloride 0.68 mg, cincocain hydrochloride 0.02 mg), Xylocaine 10% aerosol (lidocaine 10 percent) were evaluated in terms of effectiveness in decreasing the intra-oral injection pain. Six groups each consisting of 20 children were constituted from 120 children aged between 10-15 years. The responses of the patients to the pain were evaluated using a visual analogue scale. Consequently, of all the other anesthetic agents used in the present study, Vision Gel was observed to be the most effective. PMID- 10686869 TI - The effect of physiological root resorption on the histological structure of primary tooth pulp. AB - The aim of this study was to determine, the effect of physiological root resorption on the histological structure of healthy primary tooth pulp. Fourteen canine teeth, which needed to be extracted for orthodontic purposes and in which resorption had just begun (1st group, resorption did not exceed 1/3 of root length) or was in advanced resorption stage (2nd group, resorption was between 1/2 and 2/3 of root length), were used for this study. After the extraction of the teeth, they were prepared for histological examination. Then the samples were examined using light microscopy. The result no difference was found which could be detected by polarized light microscope that was related to physiological resorption and histological structure of primary teeth pulp. PMID- 10686870 TI - The effect of physiological root resorption on repair potential of primary tooth pulp. AB - The aim of this study was to determine, the effects of root resorption on repair potential of healthy deciduous tooth pulps. Fourteen canine teeth which needed to be extracted for orthodontic purposes and in which resorption had just begun (1st group, resorption did not exceed 1/3 of root length) or was in advanced resorption stage (2nd group, resorption was between 1/3 and 2/3 of root length) were used for this study. Direct pulp capping treatment was implemented in vivo, to 7 teeth in each group. Reparative dentin formation was determined three months later following extraction. The teeth were examined histopathologically under light microscope. As a result, in the teeth with different resorption levels, dentin bridge formation in the capping area was observed. Although maturation of the thin dentin bridges was completed in all teeth, maturation of the thick dentin bridges was still continuing at the 90th day. PMID- 10686871 TI - Diagnosis of dental caries: a comparison of three radiograph viewing techniques. AB - Interproximal caries scores for identical surfaces were compared across the three visualization techniques. First, the scores obtained by the naked eye method were compared to the scores obtained using the magnified view box. The second comparison looked at difference in scores between the magnified view box technique and the D.E.T.E.C.T. machine. A third comparison between the naked eye technique scores and the D.E.T.E.C.T machine scores was also made for each of the two operators. The comparison of caries score between the magnified view box and the digital image enhancement D.E.T.E.C.T. machine showed a more homogenous result. More caries was scored by both operators using the digital image enhancement system than with the view box technique. The difference in caries scores between the two techniques assigned for every surface by the two operators were statistically significant (p < 0.05). It is clear from this study that image enhanced interpretations of conventional radiographs is a necessary step to fine tune the caries diagnosis process. PMID- 10686872 TI - Combined effects of argon laser irradiation and fluoride treatments in prevention of caries-like lesion formation in enamel: an in vitro study. AB - The sample of this study consisted of 10 human permanent molars, which were sectioned into tooth quarters using slow speed diamond saw (Isomet), then a quarter from each tooth was assigned to one of four treatment groups: A) control (no treatment); B) argon laser only; C) argon laser plus neutral sodium fluoride for 4 minutes; D) argon laser with zinc fluoride for 4 minutes; each tooth quarter was coated with acid resistant varnish, leaving a window of 2 mm x 3 mm of sound enamel exposed. The results were that teeth treated with argon laser and then with zinc fluoride for four minutes have significantly reduced white spotting or etching. Zinc fluoride and argon laser combination are particularly effective in compensating for carbonate inclusion. It has a property of stabilizing hydroxyapetite crystal and restoring the structural defects of this crystal. Caries detection dye is a reliable diagnostic tool for white spot lesions. It reduces the false positive and false negative results by 60%, when compared with visual 16x magnification. PMID- 10686873 TI - The effect of thyroid hormone on orthodontic tooth movement in rats. AB - The major goals of this study were to determine the effects of different doses of thyroxin on the rate of orthodontic tooth movement and the force-induced root resorption. In this study fifty male Sprague--Dawley rats were divided into five groups: a normal group with no intervention; a control group in which appliances were placed and 10 ml/kg i.p./day normal saline was injected; and three thyroxin groups in which appliances were placed and 5, 10 and 20/microgram/kg i.p./day L thyroxin were administered, respectively. A fixed orthodontic appliance consisting of a 5 mm closed-coil spring was ligated between the maxillary incisor and maxillary first molar to deliver an initial force of 60 gm. The results showed that administration of 20/microgram/kg i.p./day L-thyroxin significantly increased the amount of orthodontic tooth movement (p < 0.001). The extent of root resorption as seen from scanning electron micrographs decreased with thyroxin administration. PMID- 10686874 TI - Transposition of mesial and distal aspects of maxillary first molars: case report. AB - Congenital absence of one or more teeth, hypodontia, is the most common developmental anomaly and is often accompanied by the presence of other tooth anomalies. In this case two Japanese sisters have several congenitally missing primary and permanent teeth and morphological abnormalities of maxillary first molars. One sister has transposition of mesial and distal aspects of a maxillary first molar, whose cusps display a normal shape. Another sister has maxillary first molars, which look like maxillary second molars. Mesio-distally shift of teeth is a very rare anomaly making this particular case important to analyze the teeth formation and development. PMID- 10686875 TI - Macrodontia in association with a contrasting character microdontia. AB - The dental, genetic, radiological and dermatoglyphic findings of a 19-year-old girl showing macrodontia of maxillary permanent central incisors in association with a contrasting character, microdontia of maxillary permanent lateral and mandibular primary central incisors and bilateral absence of maxillary first premolars and missing of the right mandibular second premolar and peg-shaped mandibular primary lateral incisors and canines were presented. PMID- 10686876 TI - Congenital lower lip pits: case report and review of literature. AB - A case of congenital lower lip pits is presented. The main characteristics of these malformations, the importance of the diagnosis and the surgical treatment are discussed. PMID- 10686877 TI - Extensive, traumatic fractures of the orbit in war and peace time. AB - Traumatic fractures of the orbit are fairly frequent injuries both in wartime and in peacetime. Most often they are part of facial injuries but they can also be isolated. They are followed by numerous symptoms such as eye dislocation, enophthalmos, double vision, and often by various injuries of eye accessory organs. The conservative treatment of these injuries has been mostly abandoned. Surgical therapy--that is, the reconstruction of the original orbital shape and volume as well as repositioning of orbital contents to their previous position- has to be performed as soon as possible. In cases of neglected fractures it is necessary to refracture the bone, reposition and fix the broken orbital fragments, and then, concomitantly, reconstruct the orbital shape and volume as well as reposition the orbital contents in their original position. In patients with neglected injuries, results of surgical treatment (i.e., correction of eye position, enophthalmos, and double vision) are considerably poorer than those achieved after the surgical treatment of fresh injuries. PMID- 10686878 TI - Temporomandibular joint biomechanical restrictions: the fluid and synovial membrane. AB - The authors analyze the functions of the synovial membrane and the chemical physical properties of synovial fluid. In particular they evaluate the role played by synovial fluid in the complex mechanism of the temporomandibular joint. Every single part that belongs to the temporomandibular joint, together with the stomatognathic apparatus, plays a specific and particular role according to the dynamics and to the preservation of the correct temporomandibular joint physiology. The physiological postural and functional relationship between the various parts of the temporomandibular joint is guaranteed by a number of biomechanical restrictions that lead and influence the regular execution of the articular movements. The most involved biomechanical restrictions in the temporomandibular joint are the temporomandibular ligament, the lateral disc ligament, the bilaminar zone or retrodiscal tissue, the synovial membrane, and the synovial fluid. PMID- 10686879 TI - Histological examination of regenerated bone through craniofacial bone distraction in clinical studies. AB - The process of bone formation by distraction osteogenesis of the craniofacial bone has been studied in animals. To our knowledge there are no published findings in which histological examination of craniofacial distraction in humans has been reported. Specimens were obtained from 10 patients who underwent craniofacial distraction: 2 patients who underwent mandibular distraction, 7 patients who underwent midface distraction, and 1 patient who underwent nasal bone distraction. These specimens were examined histologically. The results revealed that 8 of 10 patients exhibited new bone formation. No cartilaginous callus formation was observed in any of the specimens, which strongly suggests that new bone was produced by intramembranous ossification during human craniofacial bone distraction. PMID- 10686880 TI - Velopharyngeal changes after maxillary advancement in cleft patients with distraction osteogenesis using a rigid external distraction device: a 1-year cephalometric follow-up. AB - The effect of maxillary advancement on speech may have benefits on articulation improvement but compromises velopharyngeal (VP) closure by increasing the nasopharyngeal distance. The purpose of this study was to evaluate the static VP anatomic changes on lateral cephalograms in patients who underwent maxillary advancement through distraction osteogenesis (DO) with a rigid external distraction device and to correlate these changes with clinical speech data. Twenty-two patients (5 female and 17 male) underwent maxillary advancement through DO utilizing a rigid external distraction device (age, 5.2 to 25.7 years) with various diagnoses, including 13 unilateral cleft lip and palate (CLP) patients, 5 bilateral CLP patients, 1 isolated cleft palate patient, 2 facial cleft patients, and 1 patient with craniosynostosis. Lateral cephalograms of preoperative, immediate postdistraction, and 1-year postdistraction were obtained for analysis. Speech evaluation was performed preoperatively, immediate postdistraction, and then at 6-month intervals, and included assessment of air pressure flow, hypernasality, and articulation. With an average amount of 8.9 mm maxillary forward advancement, 14% of patients (3 of 21) presented deterioration in hypernasality. However, 57% of patients (12 of 21) demonstrated improvement in articulation. The cephalometric analysis demonstrated an increase in nasopharyngeal depth by 8.5 mm (1:1 ratio with bony movement) and velar angle by 14.1 deg. The length of the soft palate remained unchanged. The need ratio (intersection of palatal plane and posterior pharyngeal wall-posterior nasal spine/posterior nasal spine--tip of uvula) worsens after distraction. The deterioration of hypernasality was related to the amount of forward distraction, especially in patients without a preexisting pharyngeal flap (PF). Speech evaluation is an important aspect concerning treatment planning for maxillary distraction. The increase in nasopharyngeal depth may compromise VP closure. The increase in velar angle was considered to be part of the compensation in the VP mechanism. An adverse effect of a preexisting PF on maxillary distraction was not observed; however, it prevented postoperative hypernasality. PMID- 10686881 TI - The effect on facial growth of pediatric mandibular fractures. AB - The incidence of facial fractures in the pediatric population is between 1.4% and 15% of all maxillofacial traumas. Forty-one percent of pediatric facial fractures involve the mandible. No study has commented on the incidence of mandibular fractures that go on to develop growth disturbances leading to asymmetry and malocclusion. A retrospective chart review was carried out that identified and followed 88 children who sustained mandibular fractures and presented to The Hospital for Sick Children in Toronto during the 13-year period from 1980 to 1993. Patient follow-up ranged from 2 to 15 years, and was performed via phone survey and medical/orthodontic chart review. Patients who required orthodontics and orthognathic surgery were identified. Results indicated that a pediatric mandibular fracture does not lead to a higher incidence of orthodontic intervention. Furthermore, children younger than 4 years or older than 12 years rarely require orthognathic surgery to correct facial growth disturbances following mandibular fractures. In contrast, 22% of children age 4 to 7 years, and 17% of children age 8 to 11 years required orthognathic surgery to correct facial growth disturbances following mandibular fractures. Condylar fractures were the most common site of mandibular fracture, and led to facial asymmetry most frequently. PMID- 10686882 TI - Frontotemporal fasciocutaneous island flap for facial aesthetic subunit reconstruction. AB - The frontotemporal fasciocutaneous island flap is a useful source of tissue for correcting aesthetic units of the face. The quality of the tissue may be enhanced, and a successful color and texture match may be achieved. This flap is based on the temporal vessel system and its own fascia. Its provides excellent venous drainage and its pedicle length and arc of rotation may be increased. The donor scar is hidden under the hair-bearing area. The frontotemporal fasciocutaneous island flap was used in patients with inferior eyelid defects, for cheek reconstruction, for providing coverage of superior and inferior lip defects, for restoring the normal anatomy of columellar defects, and for reestablishing the contour of menton defects. The frontotemporal fasciocutaneous island flap was employed successfully in 9 patients at the Hospital Gea Gonzalez. The wide treatment possibilities for the reconstruction of aesthetic units in the face with the frontotemporal fasciocutaneous island flap are illustrated. PMID- 10686883 TI - Effect of human bone morphogenetic protein 2 implant on tooth eruption in an experimental design. AB - This study evaluated the influence of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the development and eruption of the secondary dentition. Primary premolar tooth extraction sockets in 12 16-week-old felines were implanted with either rhBMP-2, in collagen sponge or with buffer/absorbable collagen sponge (ACS). Unoperated jaw quadrants served as controls. Experimental conditions were randomized between jaw quadrants in all animals. Two animals receiving rhBMP-2/ACS and buffer/ACS in two quadrants per implant were sacrificed at 4 weeks postsurgery. Ten animals receiving rhBMP-2/ACS (two quadrants), buffer/ACS implants (one quadrant), and one quadrant serving as an unoperated control were evaluated at 12 weeks postsurgery. Clinical assessments included healing, eruption patterns, and crown development. Radiographic assessments included tooth development, eruption patterns, and bone formation. Histological observations were also made from the 4-week animals. The secondary dentition remained unerupted at 4 weeks postsurgery. Histological analysis showed normal alveolar bone coronal to the erupting teeth in rhBMP-2/ACS-implanted quadrants. At 12 weeks postsurgery, all teeth were erupted without differences between quadrants. Clinically, the crowns of all teeth were normal. Radiographs suggested that teeth in rhBMP-2/ACS- and buffer/ACS-implanted jaw quadrants exhibited similar tooth development and eruption patterns as the normal control. The evidence from this study suggests that surgical implantation of rh-BMP-2/ACS in the pathway of the developing and erupting secondary dentition does not interfere with the normal development and eruption patterns of the teeth. PMID- 10686884 TI - Attachment of the deep temporal fascia to the zygomatic arch: an anatomic study. AB - It is generally acknowledged that the superficial layer of the deep temporal fascia attaches to the lateral surface and its deep layer along the medial surface of the zygomatic arch. However, Ramirez asserts that the superficial and the deep layer of the deep temporal fascia fuse consistently approximately 1 cm above the upper ridge of the arch and attach to the outer surface of the arch, blending with the superficial fascia of the masseter muscle. In this study the authors clarify the precise anatomic relations between the fascia and the zygomatic arch. Coronal sections crossing the midpoint between the zygomaticotemporal suture and the tubercle of zygoma were observed grossly and histologically in 32 hemifaces from 16 Korean adult cadavers. This study demonstrates that the superficial and the deep layers of the deep temporal fascial fuse and insert onto the superior margin of the arch in 18 dissections (56%) and insert onto the superolateral surface in 14 dissections (44%). The contacting surface of the fused deep temporal fascia to the periosteum of the zygomatic arch is less than 2 mm. The following route is safer and is recommended for reaching the zygomatic arch region: Just above the split of the deep temporal fascia, a dissection is carried through the deep temporal fasica, continuing downward to the superior margin of the arch along the undersurface of the fascia. At this spot the periosteum of the arch is dissected. PMID- 10686885 TI - Correction of unilateral cleft lip nasal deformity using the sliding sulcus procedure. AB - The nose can be conceptualized as a soft-tissue structure, the orientation of which depends externally on its osseous foundation and internally on the shape and position of its cartilaginous struts. If the soft-tissue envelope of the cleft nose can be given volumetric symmetry, if it can be positioned correctly in space, and if physical forces imposed on it by cleft pathology (i.e., abnormal muscle insertion), then the internal rearrangement or augmentation of the cartilaginous components can await definitive operation later in childhood. The sliding sulcus operation is a subperiosteal procedure designed to address the problems of division, deficiency, displacement, and distortion, taking advantage of its unique ability to mobilize tissues from the maxilla. Surgical techniques are presented with clinical examples. PMID- 10686886 TI - Congenital infiltrating lipomatosis of the face. AB - Congenital infiltrating lipomatosis of the face is a rare clinical entity. Since it was first described by Slavin and colleagues in 1989, only a few cases have been reported in the literature. A 6-year-old girl with congenital infiltrating lipomatosis of the right side of the face is presented, and treatment modalities are discussed. PMID- 10686887 TI - Transoral maxillary distraction osteogenesis of an unrepaired bilateral alveolar cleft. AB - Distraction osteogenesis has gained acceptance as a viable modality for lengthening hypoplastic skeletal structures in the maxillofacial region. A case of the application of this technique to advance the maxilla in an unrepaired bilateral alveolar cleft via a transoral approach is presented. The distraction devices were applied bilaterally to the zygomatic buttress region with the activating arms protruding from the oral cavity. A high Le Fort I osteotomy was performed under general anesthesia and, prior to distraction, the three maxillary segments were unified with an occlusal acrylic splint. Activation was begun 6 days after placement, at a rate of 1 mm per day, until the planned maxillary advancement had been achieved. An 8-week period of consolidation was allowed prior to removal of the devices. PMID- 10686888 TI - Congenital trismus secondary to masseteric fibrous bands: endoscopically assisted exploration. AB - The authors present an 18-month-old female infant with congenital trismus. Computed tomography and magnetic resonance imaging were not helpful in determining the cause. A surgical endoscope was used to explore her temporomandibular joints and temporal fossae, thus avoiding the morbidity of a bicoronal incision. The cause was bilateral fibrous bands on the anterior border of the masseter muscles. Incision of these fibrous bands led to relief of the trismus. This finding is consistent with a previously described variant of the Hect-Beals-Wilson trismus-pseudocamptodactyly syndrome. This patient, however, had no evidence of the autosomal dominant inheritance pattern nor did she exhibit pseudocamptodactyly, both of which are generally ascribed to this syndrome. Unfortunately the trismus recurred 3 months postoperatively. PMID- 10686889 TI - Numb chin syndrome secondary to metastatic breast disease. PMID- 10686891 TI - Dental group practice or 2 + 2 = synergy. PMID- 10686890 TI - Orofacial pain update. 2. AB - This article is the second of a two part series, intended to update dentists regarding some of the current issues associated with pain management in the facial region. Part One presented a look at what constitutes orofacial pain and what should be included in a comprehensive evaluation prior to treating facial pain. The second part of this article focuses on levels of education, standard of care issues, specialty status and implications on how care is delivered. PMID- 10686892 TI - Dental radiology for the new millennium. PMID- 10686893 TI - The exceptional office vs. the perfect office. PMID- 10686894 TI - Feel-based design: a reason to endorse ergonomic standards. PMID- 10686895 TI - Treatment of periodontal diseases: what treatment works, and what's new. PMID- 10686896 TI - Latex hypersensitivity and dentistry. PMID- 10686897 TI - A case report: recognizing factitious injuries secondary to multiple eating disorders. AB - This report describes the uncommon problem of a female patient diagnosed with an eating disorder, bulimia nervosa, who reported self-mutilating dental factitious behavior. The case presents a serious diagnostic and management problem. Notwithstanding the clinical appearance of the dentition, a thorough medical dental history was essential for this uncommon diagnosis. PMID- 10686898 TI - Clinical crown lengthening: restorative considerations. PMID- 10686899 TI - Patient records: growing importance. PMID- 10686900 TI - Periodontal disease and heart disease--a connection. PMID- 10686901 TI - Oral bone loss associated with menopause. PMID- 10686902 TI - Multiple supernumerary teeth: literature review and case report. AB - This paper provided information regarding the classification, etiology, prevalence, sequelae and treatment of supernumerary teeth. A case of multiple supernumeraries in a healthy pediatric patient was presented. PMID- 10686903 TI - Levi Spear Parmly: the apostle of dental hygiene. AB - Dr. Levi Spear Parmly (1790-1859), an influential American figure in the field of dental prevention, introduced flossing as the most efficient way to prevent dental disease. Dr. Parmly practiced in the United States, England and France. Many of his ideas about controlling dental diseases are still highly applicable to modern clinical practice. In this century, Dr. Charles C. Bass (of New Orleans) and Dr. Sumter S. Arnim (of Houston) rediscovered, expanded, and publicized the principles, ideals and practices of Dr. Parmly. As a result, Parmly's concepts relating to oral disease causation and control continue to influence today's dental practitioners. His seminal work and thoughts concerning preventive dentistry are presented in this biographical paper. PMID- 10686904 TI - Doc Holliday's dental chair. AB - John Henry Holliday, DDS, was one of the best-known dentists of the United States. However, little is known about his actual practice of dentistry. This article discusses a dental chair that was used by Doc and shows where it fits into his dental life. PMID- 10686905 TI - Toothaches and death. AB - Deaths from dental abscesses today are so rare, that it is difficult to fathom that only 200 years ago, this was a leading cause of death. When the London (England) Bills of Mortality began listing the causes of death in the early 1600's, "teeth" were continually listed as the fifth or sixth leading cause of death. (This does not include the category of "Teething" which was probably erroneously blamed for many children's deaths. As we examine several historic factors of this period, it is apparent that the number of deaths attributed to "teeth" in the seventeenth and eighteenth centuries was probably fairly accurate, and it was not antibiotics, nor the discovery of asepsis, that brought about the dramatic reduction in these dental mortalities, but two much earlier dental innovations. PMID- 10686906 TI - Dentistry on stamps. Tiradentes. PMID- 10686907 TI - Freud's cancer. PMID- 10686908 TI - Dr. W. Harry Archer, DDS, MA, 1905-1980 a life of dedication to oral surgery. AB - The life of the American oral surgeon Dr. W. Harry Archer was one that affected and inspired, and sometimes incensed, those who knew him and those who would follow him. His life long devotion to the dental specialty of oral surgery was a driving force that caused dramatic changes in aspects of not only oral surgery but of dentistry as a whole. His excellence in leadership, teaching, research, and writing were among his many accomplishments that firmly place him among the eternal giants of the dental community. PMID- 10686909 TI - Thomas Lea Buckingham, MD, DDS, (1816-1883). AB - Miller, farmer, mechanic, dry goods merchant, dentist, author, physician, teacher, inventor, and fourteenth president of the American Dental Association, take your pick, for Thomas Lea Buckingham without feigning, embraced that menu to the brim--the verge beyond which, only a pittance of dentists might succeed. Nevertheless, the facts lie untouched--a testament to one of dentistry's most brilliant, unassuming, and dedicated disciples. Loved by his students and respected by his peers, Thomas Lea Buckingham reached new heights in garnishing respect from his colleagues. Sharing his knowledge, Dr. Buckingham profusely wrote about his findings, being affirmed by at least 25 dental periodicals (many with multiple contributions), over a period of thirty-four years, from 1850 to 1884. His principal writings appeared in Dental Cosmos, Johnston's Dental Miscellany, New York Odontologic Society Transcripts, Odontologic Society of Pennsylvania, American Dental Association Transactions, Dental Items of Interest, Pennsylvania Journal of Dental Science, Dental News, Dental Jairus, Missouri Dental Journal, Dental Registrar, New England Journal of Dentistry, and the British Journal of Dental Science. Upon Buckingham's death, noted dentists of his era spoke with love and respect of this exceptional dentist. This paper highlights the maturation of a minimally educated miller, who, with determination and purpose, rose to the pinnacle of his selected professions and garnered the love and respect of his fellow men. PMID- 10686910 TI - Memories of oral health and dentistry in the 19th century: advertising trade cards. PMID- 10686911 TI - The explorer. PMID- 10686913 TI - Basil Manly Wilkerson: dental inventor extraordinaire. PMID- 10686912 TI - Henry Daniel Cogswell, DDS (1819-1900): a temperance advocate, philanthropist and builder of ice-water fountains. AB - Henry Daniel Cogswell (Fig. 1), the second of five children, was born in Tolland, Connecticut on March 3, 1819. His father, George Washington Cogswell, was a general carpenter, architect and builder of moderate circumstances. In 1827, when Henry was eight, his mother died. The following year, Henry's father moved to Orwell, (Oswego County) New York, in hopes of improving his financial condition. Henry was left behind in the care of his paternal grandfather, who died several months later, leaving the 10-year old boy, stranded and forced to rely upon his own resources. (In those times, when families were separated, individual members had limited means of locating one another.) PMID- 10686914 TI - Tobacco and oral health. PMID- 10686915 TI - Gleanings about dentistry from the world of literature (eighteenth in a series). PMID- 10686916 TI - Giuseppangelo Fonzi: industrial fabrication promoter of porcelain prosthetics. PMID- 10686917 TI - Veterinary dentistry: its origin and recent history. PMID- 10686918 TI - Obituary: Norman Harry Olsen, DDS, MSD. PMID- 10686919 TI - Survival of the fittest. PMID- 10686920 TI - Update in periodontology. PMID- 10686921 TI - The periodontal aspect of anterior aesthetics. PMID- 10686922 TI - Osseointegrated implant failures. AB - This article discusses the criteria used for implant success and failure, the classification of implant failures, the causative factors, and diagnosis of the failing and failed implant. In spite of the impressive success rates of osseointegrated dental implants, failures occur and in some studies the incidence of failure is high. Many studies do not use objective criteria to define success and confuse survival with success. The criteria used affect reported success rates. Implant failures may occur early (primary) after implant placement or after the implant is loaded (secondary). There is no single aetiological factor and failures have been attributed to poor surgical technique, host factors that impair healing, poor bone quality, peri-implant infections, poor prosthesis design and traumatic loading conditions. Early diagnosis of problems is critical and every effort should be made to treat the problem while the damage can still be managed or even reversed. PMID- 10686923 TI - Enhancement of bone ingrowth into collagen/HA composite implants using e-PTFE membranes. AB - The objective of this investigation was to study the effectiveness of expanded polytetrafluoroethylene (e-PTFE) membranes for enhancement of bone ingrowth through subperiosteally implanted collagen/HA composite blocks. Twelve rabbits aged 12-15 months served as the experimental animals in this study. Two compressed Collagen/HA composite blocks in the shape of two attached cylinders of different diameters were inserted into two defects of each rabbit calvarium, the smaller cylinder being intrabony, the larger subperiosteal in location. One of the two implants was covered with non-resorbable e-PTFE membrane. The other implant was left uncovered. Specimens were obtained at 4, 8, and 12 weeks. While the implant specimens on the membrane side showed progressive bone formation between and around HA particles at the subperiosteal extrabony locations, the HA particles on the non-membrane side were surrounded and separated by dense fibrous tissue. At intraosseous sites, HA particles were surrounded by new bone throughout the defect on the membrane side, but new bone formation occurred only along the periphery on the non-membrane side. It appears that guided tissue regeneration may be used to enhance new bone formation around and between subperiosteally implanted HA particles. PMID- 10686924 TI - Core build-up materials and techniques. PMID- 10686925 TI - Restoration of endodontically treated teeth with posts and cores. AB - This article is a review of the current technique and materials used in the restoration of endodontically treated teeth with post and core materials and procedures. It is intended to aid clinicians in making selections which are appropriate to their individual practices through the evaluation of relevant literature about the restoration of endodontically treated teeth. PMID- 10686926 TI - The Catholic University of Ireland (1854-1908). PMID- 10686927 TI - Performance evaluations--three sides of the coin. PMID- 10686928 TI - Mercury amalgam safety: a review. PMID- 10686929 TI - Exposure incidents: protective measures and action strategies. PMID- 10686930 TI - Prosthetically determined implant placement for the partially edentulous ridge: a reality today. PMID- 10686931 TI - Management of occupational exposures to bloodborne viruses: risk and risk factors for disease transmission. PMID- 10686932 TI - Are we meeting the oral health needs of our pediatric patients? PMID- 10686933 TI - Changing perspectives in the management of the preschool child. PMID- 10686934 TI - Advantages of new restorative materials in dental care for children. PMID- 10686935 TI - The vital pulpotomy in primary molars. AB - The vital formocresol pulpotomy is a useful technique to regain satisfactory oral health. Proper evaluation and diagnosis is crucial in order for the dentist to select the pulpal therapy that offers the best chance of long-term success with the fewest complications. PMID- 10686936 TI - Mutation analysis of BRCA1, TP53, and KRAS2 in ovarian and related pelvic tumors. AB - Cancer may be viewed as a genetic disease resulting from critical mutations that disrupt normal cell growth. To characterize the involvement of the BRCA1 and TP53 tumor suppressor genes and of the KRAS2 protooncogene in gynecologic cancer, mutation analysis of these genes was conducted in pelvic tumors of 85 patients that included 49 epithelial ovarian carcinoma cases. The 85 pelvic tumors contained 5 tumors with BRCA1 mutations, 33 with TP53 mutations, and 1 with a KRAS2 mutation. Each of the BRCA1 and KRAS2 mutations, and 25 of the TP53 mutations, were in ovarian carcinomas. Four of the BRCA1 mutations were germline and 1 was somatic. The 4 patients with germline BRCA1 mutations had an early age of disease onset (33-48 years) relative to the mean age of onset (58 years) of all 49 ovarian carcinoma patients, and 3 of these 4 patients had a family history of ovarian or breast cancer. None of the 4 tumors with germline BRCA1 mutations had a KRAS2 mutation or a TP53 mutation, despite a 51% frequency of TP53 mutations in the 49 ovarian carcinomas. Three of the 4 tumors with germline BRCA1 mutations retained a wild-type BRCA1 allele. The tumor with the somatic BRCA1 mutation contained a TP53 mutation and had no evidence for wild-type BRCA1 and TP53 alleles. These data suggest that both BRCA1 and TP53 were inactivated in 1 of 49 ovarian carcinomas. Moreover, mutational inactivation of both BRCA1 and TP53 did not occur in 4 tumors with a germline BRCA1 mutation. It has been proposed that tumorigenesis in cells with a heterozygous BRCA1 mutation requires inactivation of the wild-type BRCA1 and TP53 alleles, which results in genomic instability and acquisition of mutations in protooncogenes. Clearly, mutational inactivation of TP53 and the wild-type BRCA1 allele in ovarian tumors with a heterozygous, germline BRCA1 mutation is not an absolute requirement for tumor formation. It is possible that these alleles may be inactivated by nonmutational mechanisms or that other tumor formation pathways exist. PMID- 10686937 TI - Coexistence of several unbalanced translocations in a case of neuroblastoma: the contribution of multicolor spectral karyotyping. AB - Spectral karyotyping (SKY) is based on the simultaneous hybridization of a set of 24 chromosome-specific DNA painting probes, each labeled with a different fluor combination. Automatic classification, based on the measurement of the spectrum for each chromosome, was applied to metaphases obtained from the affected bone marrow of a neuroblastoma case. Spectral karyotyping allowed the identification of chromosomal aberrations that could not be identified by the use of the G banding technique, and revealed a number of gains and unbalanced translocations. PMID- 10686938 TI - Phenotypic and genotypic diversity of human neuroblastoma studied in three IGR cell line models derived from bone marrow metastases. AB - Metastatic stage IV neuroblastoma tumors, as well as cell lines derived from them, are highly malignant and rapidly fatal. To determine whether malignant potential of these cells might be influenced by stromal tissue at sites frequently involved in metastasis, we initiated primary cultures from bone marrow of three patients (331, 337, and 91) with stage IV neuroblastoma. All three explants contained two distinct cell populations, malignant neuroblasts (Nb-type) and substrate adherent stromal-like (Str-type) cells. The cell types were separated at the first passage and studied by cytogenetic, molecular, and immunocytochemical methods. Karyotypic analyses after 3-6 passages in vitro revealed the presence of unique chromosomal abnormalities in Nb-type cells of all three lines: (1) der(1)t(1;7) (p32;q11) and der(5)t(5;17)(q35;q21) in pseudodiploid IGR-N-331 neuroblasts; (2) der(1)t(1;17)(p35;q21-22) x 2 and der(7)t(7;7)(p21;q21) in IGR-N-337 hyperdiploid neuroblasts; and (3) more than six rearranged chromosomes in two related subpopulations of hypodiploid IGR-N-91 neuroblasts. Neuroblastic cells from all three tumors amplified MYCN 25- to 50 fold (with amplified genes visible as dmin or, in one IGR-N-91 subline, as an hsr(14)[q32]) and expressed N-CAM. Str-type cells from tumors 331 and 337 had a normal diploid karyotype, did not express either N-CAM or S-100, and are probably normal bone marrow fibroblasts. By contrast, S-100 negative Str-type IGR-N-91 cells were hypodiploid and shared at least two unbalanced translocations, der(4)t(1;4)(q12;p15) and der(2)t(2;10;17)(p14;q11;q22), with neuroblastic counterparts, indicating that "stromal" cells and malignant neuroblasts had a common tumor cell origin. Thus, the Str-type cells of IGR-N-91 are examples of S type phenotypic variants frequently described for long-term human neuroblastoma cells lines in vitro, but not previously observed in vivo. PMID- 10686939 TI - Atypical chronic myeloid leukemia with der(20)t(17;20)(q21;q13). AB - We present a case of atypical chronic myeloid leukemia that developed blastic transformation four months after initial presentation, with the blast cells showing multilineage antigen expression. A novel karyotypic abnormality, der(20)t(17;20)(q21;q13), was found in bone marrow cells at diagnosis. The potential role of such an aberration in leukemogenesis is discussed. PMID- 10686940 TI - Genetic studies on a family with acute myelogenous leukemia. AB - Seven cases of myelogenous leukemia--two acute erythroleukemia (AEL), four acute myelogenous leukemia (AML), and one acute myelomonocytic leukemia (AMMoL)--were found in 22 members of three consecutive generations of a family in the past 16 years (1973-1989). By using cytogenetic, hematologic, and biochemical analyses of those surviving in this family, we also found four members who might develop leukemia in the future. Southern blot analysis of one of the four members and her father (an acute leukemia patient) with a v-ERBB probe showed that the gene abnormalities consisted of a c-ERBB rearrangement (hereditary) and a rearrangement/amplification of the same gene. PMID- 10686941 TI - Trisomy 1q generating translocations in Wilms tumor. AB - Unbalanced translocations generating trisomy of 1q are common in Wilms tumor (WT). We present eight unbalanced 1q translocations from seven tumors and a review of the literature. An unbalanced translocation that results in a der(16)t(1q;16q) chromosome represents more than half of the published +1q generating translocations in WT. This translocation is also common to many other tumor types. Four of the tumors presented here contained this chromosome and,in two cases, it was the primary acquired cytogenetic abnormality within the tumor. The other four translocations involved 9q31, 9q34, 17p1?, and 21p11 as the partner to 1q. The chromosome 17 and 21 translocations occurred within the same tumor as apparently independent events. In contrast with the 16q translocations, these other translocations were secondary cytogenetic events, thereby indicating a role in tumor progression rather than initiation. Probes mapping to 1q12 and 1q21 were employed for fluorescence in situ hybridization and it was demonstrated that different 1q breakpoints are possible. In this series, the majority of breakpoints either mapped to 1q12 or were centromeric to this region. PMID- 10686942 TI - Characterization of the C-MYC amplicon in a case of acute myeloid leukemia with double minute chromosomes. AB - We have characterized the double minute chromosomes in a case of acute myeloid leukemia (AML). Southern blot analysis showed that the C-MYC was amplified. Further analysis with probes located both 3' and 5' of MYC indicated that the amplicon was at least 700 kb in size, extending from the papilloma virus integration site situated 500 kb 5' of MYC to the PVT gene located 280 kb 3' of MYC. This appears to be the largest MYC-containing amplicon in human leukemia. PMID- 10686943 TI - Unusual chromosome patterns of renal cell carcinomas common to two brothers. AB - In this study, we describe two renal cell carcinomas (RCC) that occurred at the same time in two brothers, yielding information on the carcinogenic process. We used flow cytometry (FCM) to evaluate nuclear DNA content, and performed cytogenetic analysis. We also carried out fluorescence in situ hybridization (FISH) with a panel of centromeric probes for chromosomes 3, 7, 8, 9, 12, 17, 20, and Y in interphase cells. Flow cytometry analysis revealed diploid histograms in the tumor and "nonmalignant" samples of patient 1, while an aneuploid cell subpopulation was found in the tumor and "nonmalignant" samples of patient 2. Tumor samples from the two brothers were studied by FISH, and had common numerical chromosome aberrations: trisomy of chromosomes 3 and 7, and monosomy and trisomy of chromosomes 9 and 17. Moreover, in normal samples from both brothers, we found monosomy 9, and in a normal sample from patient 1, monosomy 17. Cytogenetic analysis revealed trisomy 3 in some cells grown from normal kidney tissue of each brother. The identification of the same chromosome alterations in both brothers appears to provide evidence of an unusual process of carcinogenesis, probably due to a common genetic basis. PMID- 10686944 TI - Inflammatory myofibroblastic tumor with HMGIC rearrangement. AB - Inflammatory pseudotumors or inflammatory myofibroblastic tumors (IMT) are lesions of extreme heterogeneity showing a highly variable mixture of bland looking spindle cells, inflammatory cells, and collagen fibers. We describe our results of molecular cytogenetic and rapid amplification of cDNA ends (RACE-PCR) studies on an IMT characterized by a translocation involving 12q15. Chromosomal aberrations involving this region are very frequent among other benign tumors, such as lipomas, uterine leiomyomas, or pulmonary chondroid hamartomas. Recently, we have shown that, by these structural chromosomal aberrations, the HMGIC gene is affected. Fluorescence in situ hybridization (FISH) analysis and 3' RACE-PCR on cells of the present case of an inflammatory myofibroblastic tumor indicated an intragenic rearrangement of HMGIC, resulting in an aberrant transcript of that gene. Clonal cytogenetic aberrations have been described in very few cases of IMT. The results presented herein indicate that this case of IMT represents a true benign mesenchymal neoplasm associated with, or due to, a rearrangement of HMGIC. PMID- 10686945 TI - SiMa, a new neuroblastoma cell line combining poor prognostic cytogenetic markers with high adrenergic differentiation. AB - We describe the establishment and characterization of a new neuroblastoma (Nb) cell line, SiMa, carrying the major recurrent chromosome changes associated with poor prognosis Nb, including amplification of N-MYC by formation of double minutes (dmin), der(1)t(1;17)(p35;q12) and der(22)t(17;22)(q22;p13), and loss of chromosome 11, documented at both initiation and late passage. In contrast to these cytogenetic stigmata of poor prognosis, analysis of catecholamine synthesis by high pressure liquid chromatography (HPLC) measurement revealed an advanced degree of adrenergic differentiation with high rates of 3,4 Dihydroxyphenylalanine (DOPA), noradrenaline, homovanillic acid (HVA), and vanillylmandelic acid (VMA) production. Contrastingly advanced differentiation and poor prognostic genetic markers combine to render SiMa a unique instrument for investigating the pathology and therapy of Nb. PMID- 10686946 TI - A man with natural killer cell lymphoma showing 46,XX and deletion 6q. AB - Specific chromosomal abnormalities have been shown to be associated with certain types of leukemia and lymphoma. We and others have recently demonstrated del(6)(q21q25) as being strongly associated with natural killer cell lymphoma/leukemia. In this report, we describe a case of natural killer cell lymphoma with a clonal chromosomal abnormality of 46,X,-Y,+X,t(2;9)(q31;p24), del(4)(q21q25),del(6)(q21q23), and propose that the region 6q23 is probably an important site of genetic alteration in this group of tumors. PMID- 10686947 TI - DNA copy number losses at 1p32-pter in monozygotic twins concordant for breast cancer. AB - To find similarities that may possibly indicate novel mutations, we performed comparative genomic hybridization (CGH) analysis following degenerate oligonucleotide primed polymerase chain reaction (PCR) for DNA obtained from unique material of breast cancer that developed in monozygotic twin-pairs. Polymerase chain reaction amplification was successful in 12 samples for 11 patients, including 3 pairs. Six samples exhibited DNA copy number changes. Gains (76%) were more frequent than losses (24%). Gains or high-level amplifications in 8q were present in all but 1 of the abnormal cases. Frequent gains were detected with a minimal common overlapping region at 5p (4 cases), at 1q25-qter (3 cases), and at 20q12-qter (2 cases). The most frequent loss, detected in half of the abnormal cases, was at 1p32-pter. One twin-pair showed similar changes in 4 chromosomal locations involving loss of 1p32-pter and gains in 1q25-qter, 5, and 8q. PMID- 10686948 TI - A novel chromosomal rearrangement associated with therapy-related acute leukemia. AB - We describe a 7-year-old girl with therapy-related acute myeloid leukemia (AML) associated with a single and novel karyotypic abnormality. The patient had been treated with alkylating agents and etoposide for hypothalamic pilocytic astrocytoma at age 17 months, and developed mixed lineage AML. Cytogenetic analysis of the leukemic blasts showed 46,XX,der(7)t(7;11)(q22;q14) in all cells examined. Southern blot analysis revealed three copies of an unrearranged MLL gene on chromosome 11q. This is the first report of a triplicated, unrearranged MLL gene in association with a deletion of 7q anomaly and an unbalanced translocation in therapy-related leukemia. PMID- 10686949 TI - Translocations (X;10)(p22;q24) and (1;10)(q21;q11) in a follicular adenoma of the thyroid without apparent involvement of the RET protooncogene. AB - We report here the cytogenetic analysis of a follicular adenoma of the thyroid which revealed an abnormal clone with a t(X;10)(p22;q24) and a t(1;10)(q21;q11) together with normal cells. Fluorescence in situ hybridization (FISH) with YACs 273E3 and 344H4, which are located on 10q11.2 and are specific for the RET protooncogene, showed no abnormalities. It would therefore appear that this gene is not involved in the particular tumor, as has been reported in a number of papillary thyroid carcinomas. Several chromosomal aberrations have been suggested as been specific for follicular thyroid adenoma. However, until now, only a few such cases have been reported which involve structural abnormalities of chromosomes 10q11.2 and 10q24. We believe this to be the first report of a follicular thyroid adenoma with a t(X;10)and a t(1;10). PMID- 10686950 TI - Detection of MLL gene self-fusions by RT-PCR and automated fluorescent DNA fragment analysis. PMID- 10686951 TI - Constitutional (5;18) in a patient with aplastic anemia. PMID- 10686952 TI - Translocation (5;19)(q33;q13) as the sole abnormality in a case of refractory anemia with excess of blasts in transformation. PMID- 10686953 TI - Second case of t(6;7)(p21;q36) in acute lymphoblastic leukemia. PMID- 10686954 TI - Complex rearrangement of chromosomes 1, 7, and 16 in chronic myelomonocytic leukemia. PMID- 10686955 TI - [The correlation between the CD4 count in HIV-positive patients and the radiological findings in diseases of the paranasal sinuses]. AB - BACKGROUND AND OBJECTIVE: Sinusitis is a frequent and typical disease in HIV positive patients. It was the aim of this study to examine retrospectively computed tomograms (CT) and magnetic resonance images (MRI) of such patients for sinusitis. PATIENTS AND METHODS: CT and MRI of the nasal sinuses were retrospectively analysed in 71 HIV-positive patients. RESULTS: Sinusitis was diagnosed in 49 of the 71 patients by conventional radiology (38), CT (13) and/or MRI. In patients with sinusitis, contrary to those without it, there was a significant correlation between progression of the HIV infection, as measured by reduction in CD4 count, and an increased number of symptoms (r = -0.297; p < 0.05). The latter also correlated significantly with the frequency of concomitant sinusitis (r = 0.336, p < 0.05). The radiological findings correlated more closely with the CD4 count when tomography was used. The correlation coefficient for the number of patients with concomitant sinus disease rose from r = -0.305 to r = -0.459 and for the degree of severity from r = -0.324 to r = -0.484 (p < 0.05). Severity of sinus disease was clearly more marked among HIV-positive than HIV-negative patients. CONCLUSION: The severity of sinusitis in HIV-positive patients correlated directly with the level of the CD4 count. In these patients tomographic methods are preferable to conventional radiology. PMID- 10686956 TI - [Bougie endosonography in the preoperative diagnosis of stenosing esophageal carcinomas]. AB - BACKGROUND AND OBJECTIVE: Nowadays ultrasonic endoscopy is accepted as the most accurate method in the preoperative TNM staging of oesophageal tumour. At the time a diagnosis of oesophageal is made, 25-62% of patients have marked oesophageal stricture due to an advanced stage of the tumour. As a result, it is often impossible to pass a conventional endoscope and the examination is incomplete or a dilator has to be introduced at high risk of perforation or other complications. An ultrasonic oesophagoprobe (Olympus MH 908) with a diameter of 7.9 mm, its tip acting as a dilator has been developed so that even in high-grade oesophageal stricture a complete and low-risk investigation can be undertaken. The value of this instrument has been compared prospectively with that of a standard ultrasonic endoscope in patients with oesophageal stricture due to carcinoma and in relation to the postoperative histology. PATIENTS AND METHODS: Between May 1996 and February 1997, 62 patients (55 men, 7 women; average age 55.8 [41-82] years) with suspected or histologically confirmed oesophageal carcinoma were examined with the standard ultrasonic endoscope and the new oesophagoprobe. The two independent results were related to the postoperative histology. RESULTS: It was possible to compare the endoscopic results with the postoperative histology in 47 patients. In 55.8% it was not possible to pass the stricture with the standard ultrasonic endoscope (SE) so that an accuracy of only 41% was obtained in the T stage and 56.4% in the N stage. But with the oesophagoprobe (OP) an accuracy of 74.5% in the T stage and of 63% in the N stage were achieved. The difference in the findings between the two instruments was highly significant (p < 0.001) in those patients in whom the SE could not be passed through the stricture. For the T stage the accuracy was 14.3% vs. 76.2% (SE vs. OP); in the N stage it was 38.1% and 57.2%, respectively. When both instruments could be passed the results were comparable. CONCLUSIONS: Examination with the ultrasonic oesophagoprobe is a reliable and accurate method free of complications for the preoperative assessment of oesophageal carcinoma with stricture. PMID- 10686957 TI - [Retroperitoneal desmoid tumor with kidney failure in familial adenomatous polyposis]. AB - HISTORY AND FINDINGS: A 39-year-old man was hospitalized because of continually rising urinary creatinine and blood urea nitrogen concentration. He was known to have familial adenomatous polyposis (FAP), first diagnosed 18 years previously and re-Physical examination was unremarkable except for pain on percussion over both kidney regions. There was a well-healed laparotomy scar. INVESTIGATIONS: Ultrasound revealed chronic bilateral obstructive renal disease, grade II-III, and computed tomography showed a conglomerate retroperitoneal tumour with obstruction of both ureters at the level of the lower pelvis. This tumour had first been noted first 3 years after the colectomy when the patient complained of abdominal pain. It had been identified histologically as a nonresectable retroperitoneal desmoid tumour. TREATMENT AND COURSE: An external fistula was made, relieving the renal retention. To suppress growth of the desmoid tumour Sulindac, a nonsteroid anti-inflammatory drug, was administered. Genetic molecular analysis revealed a germ line defect in codon 1690 of the APC gene. It is intended to examine other members of the family for the presence of this defect. CONCLUSION: Desmoid tumours are more common in persons with FAP and are among the most frequent extracolic causes of their death. Treatment options are critically analysed. PMID- 10686958 TI - [Alveolar echinococcosis: therapy]. PMID- 10686959 TI - [Peripartal cardiomyopathy--its pathophysiology, diagnosis and current therapy]. PMID- 10686960 TI - [With chemistry against syphilis--the beginnings of chemotherapy around Paul Ehrlich and the DMW. Deutsche Medizinische Wochenschrift]. PMID- 10686961 TI - [Procalcitonin in pediatric emergencies: comparison with C-reactive protein, interleukin-6 and interferon alpha in the differentiation between bacterial and viral infections]. AB - OBJECTIVE: Procalcitonin concentration increases in bacterial infections but remains low in viral infections and inflammatory diseases. The change is rapid and the molecule is stable making it a potentially useful marker for distinguishing between bacterial and viral infections. PATIENTS AND METHODS: Procalcitonin (PCT) was determined with an immunoluminometric assay on plasma collected at admission in 436 infants and children hospitalized for bacterial or viral infection. It was compared with C reactive protein, interleukin-6 and interferon-alpha measured on the same sample. RESULTS: PCT was 41.3 +/- 77.4 micrograms/l in children with septicemia or bacterial meningitis (n = 53), 0.39 +/- 0.57 microgram/l in children with viral infection (n = 274) and 3.9 +/- 5.9 micrograms/l in children with a localized bacterial infection who had a negative blood culture (n = 109). PCT was > 1 microgram/l in 126 children with a localized or systemic bacterial infection (sensitivity 78%). PCT was < 1 microgram/l in 258 children with a viral infection (specificity 94%). For differenciation between viral and bacterial infections, CRP value > or = 20 mg/l, IL-6 > 100 pg/ml and interferon-alpha > 0 Ul/ml have 85, 48 and 76% sensitivity and 73, 85 and 92% specificity respectively. CONCLUSIONS: In this study, a PCT value of 1 microgram/l or greater had better specificity, sensitivity and predictive value than CRP, IL-6 and interferon-alpha in children for distinguishing between viral and bacterial infections. PCT may be useful in pediatric emergency room for making decision about antibiotic treatments. PMID- 10686962 TI - [Leukemia and pre-leukemic conditions occurring after treatment of breast cancer]. AB - OBJECTIVE: The purpose of this study was to determine the clinical and prognostic features of leukemias and preleukemic states, whatever the mode of development, observed in patients after treatment of breast cancer. PATIENTS AND METHODS: A retrospective multicentric analysis was made of 121 patients treated for breast cancer and who later developed leukemia or a preleukemic state. Initially, 44 patients had undergone mastectomy, 72 had conservative surgery and 119 had locoregional irradiation. At least one chemotherapy session was performed in 90 patients and 48 had received tamoxifen. The risk of relapse of breast cancer was high, moderate or low for 44, 46 and 24 patients respectively (data not available for 7 patients). RESULTS: By class, the hematology diseases found were: myelodysplasia (n = 9), refractory anemia with blast excess (n = 7), acute lymphoblastic leukemia (n = 6), acute myoblastic leukemia (n = 93 including a majority of type 2 and type 4). For acute myeloblastic leukemia, mean delay to onset was 65 and 37 months respectively without and after chemotherapy. The prognosis of these cases of leukemia and preleukemic states was poor with an overall death rate of 86%. CONCLUSION: In light of the recent development of indications for adjuvant chemotherapy even for subgroups of patients at moderate risk, it is important to more precisely assess the absolute benefit in terms of survival compared with the risk of severe complications, particular secondary leukemia. In the future, a systematic registry and a case-control study are required. PMID- 10686963 TI - [Severe gastrointestinal hemorrhage secondary to diffuse angiodysplasia: efficacy of estrogen-progesterone treatment]. AB - BACKGROUND: Vascular abnormalities are being reported with increasing frequency as a cause of major lower gastrointestinal hemorrhage in the elderly. They are occasionally very difficult to treat by conventional means. CASE REPORT: A 66 year-old white man with a history of type 2 diabetes mellitus, coronary artery disease, congestive heart failure, severe peripheral arterial occlusion disease and chronic renal insufficiency presented for five years recurrent major bleeding due to gastrointestinal angiodysplasia, requiring repeated transfusions. He was treated with efficacy using ethinyl-estradiol (30 micrograms) and norethisterone acetate (1 mg) given orally once daily. After six months of treatment, transfusion requirements fell to 0 unit and the patient's hemoglobin was stable at 13 g/dl. Attempts to stop hormone therapy (by the patient himself, without complaint of side effects) led to a fall in hemoglobin. CONCLUSION: Hormonal therapy should be considered when multiple degenerative mucosal vascular bleeding lesions are beyond the reach of therapeutic endoscopy leading to high transfusion needs and when surgical risk is unacceptably high. PMID- 10686964 TI - [Glucose-6-phosphate dehydrogenase deficiency and hemoglobinuric biliary fever after taking mefloquine]. PMID- 10686965 TI - [Wide QRS: a criterion of imminent severity of beta blockader poisoning]. PMID- 10686966 TI - [Primary antiphospholipid syndrome disclosed by acute abdominal pain]. PMID- 10686967 TI - [The RALES study]. PMID- 10686968 TI - [HIV infection in Africa. Clinical and therapeutical research]. AB - A MAJOR HEALTH PROBLEM: Human Immunodeficiency Virus (HIV) infection is a major public health problem in sub-Saharan Africa and the care of HIV-infected patients is limited by the lack of resources. Clinical research can play a major role to assess the benefit of preventive and/or curative measures adapted to the context of these countries. To illustrate advances and gaps in HIV/AIDS clinical research in Africa, we explored three issues relevant to this research: opportunistic infections in adults, mother-to-child transmission of HIV and the ethical questions. EPIDEMIOLOGY: Epidemiological African studies have shown: the omnipresence of tuberculosis, first cause of death among HIV+ patients; the frequency of bacterial infections, first cause of serious morbidity and second cause of death; the high frequency of toxoplasmosis, cryptococcal meningitis, isosporiasis, cryptosporidiasis, and other infectious syndromes of unknown etiology. More research efforts need to be done for improving tuberculosis diagnosis, compliance to treatment (evaluation of Directed Observed Therapy), resistance to treatment and primary chemoprophylaxis which has shown clear short term benefit but median term interest remains to be demonstrated. Chemoprophylaxis of opportunistic infections other than tuberculosis needs also to be evaluated: cotrimoxazole reduces the short term mortality of HIV+ patients with tuberculosis and the early serious morbidity of HIV+ patients without tuberculosis. TRANSMISSION: Mother-to-child transmission of HIV can occur during pregnancy, during delivery and the postnatal period by breastfeeding, a common practice in Africa. The overall risk of vertical transmission is estimated to be 30% but the attributable part of breastfeeding needs to be further explored. Beyond the prevention of sexual transmission of HIV among childbearing women and family planning for HIV+ women, interventions aimed to reduce mother-to-child transmission depend on the availability or not of a proposing and realising an HIV counselling and testing: antiretroviral treatments and/or breastfeeding alternatives which reduce efficaciously transmission require HIV testing, while vaginal disinfection and vitamin supplementation whom efficacy needs to be demonstrated do not. PREVENTION: Prevention of mother-to-child transmission and care of HIV+ adults in the area of opportunistic infections are feasible in Africa with an acceptable cost. This requires first to train and inform health care providers and the populations. Lots of uncertainties in these areas are likely to be alleviated by reinforcing clinical and therapeutic research of good quality including the questions of antiretroviral treatment. Ethical issues raised by the design and conduct of clinical research in Africa need a positive thinking to face the HIV African pandemic. PMID- 10686969 TI - [The 39th ICAAC (San Francisco, September 1999). HIV infection in clinical practice]. PMID- 10686970 TI - [Manic-depressive conditions in adolescents. Epidemiological and clinical aspects]. AB - EPIDEMIOLOGY CONTEXT: The prevalence of maniac depressive disorders is similar in adolescents and adults, i.e. about 1% with a 1:1 sex-ratio. Risk is higher in families with a diseased member and early episodes of mood disorders are probably correlated with the genotypic severity or the presence of a unique susceptibility gene. HIGHLY VARIABLE CLINICAL SIGNS: Until recent years, the highly variable clinical expression with rapid changes in mood, bipolar states, variable somatic, behavioral or addictive symptomatology, cognition disorders, and disturbed ideation or hallucinations, probably contributed to our poor understanding of juvenile forms of the disease. EARLY MANAGEMENT: Early diagnosis and psychiatric care is crucial due to the short-, mid- and long-term risk of unfavorable or even fatal consequences. Indeed, while still in the process of structuralization, the predisposed personality is particularly reactive to positive or negative events. It is most difficult to achieve flexibility once a restrictive organization of the personality has been installed. In addition, these families often have a painful past and lack sufficient capacity to successfully deal with the stress of emotions and conflicts occurring in the future adult during the self identification and independence-seeking processes. This familial situation points out the importance of implicating the family and close friends in the treatment strategy as a complement to drug therapy and psychotherapy proposed to the adolescent. PMID- 10686971 TI - [Manic-depressive disorders in adolescence. Mood disorders and psychoses in adolescence]. AB - DISEASE ONSET: In adolescents, the different aspects of mood disorders and psychoses are closely related. The first episode of what will become schizophrenia is often suggestive of a mood disorder. The inverse is also true. Frequently, a psychotic state is the inaugural manifestation of a mood disorder. SIGNS AND SYMPTOMS: The depressive manifestations observed in adolescents are very similar to negative psychotic symptoms. More so than in adults, the thymic disorder is expressed as severe episodes of psychosis. It would appear that in this case, the psychotic elements are related to the intensity of the thymic disorder. DIAGNOSTIC ERRORS: Misdiagnosis is probably related to the fact that mood is not sufficiently taken into account in acute psychotic states. The risk inherent in "over"-diagnosis of schizophrenic disorders is related to the therapeutic implications: prescriptions of neuroleptics can hinder the psychic work involved in the structuralization process going on in the adolescent. PMID- 10686972 TI - [Manic-depressive disorders in adolescence. Psychoanalytic point of view]. AB - CLINICAL OBSERVATIONS: Melancholy and maniac disorders do not take on their typical clinical form until adolescence, at the very time the psychic maturation and identification processes are structuring the "definitive" personality. These processes enable the subject to acquire his/her psychic independence of the primary objects, particularly the primary maternal object and secondarily the parent object. THE MELANCHOLIC OBJECT: In so-called melancholic personalities, the identification with a primary maternal object was not totally successful during infancy and had to follow a narcissistic mode. When during adolescence the subject is faced with symbolic or real separations (independence) his/her identification object has to split with loss of the invested object, leading to melancholy (or its negative pseudo-triumphant variant: maniac disorder). IMPORTANT PROCESSES IN ADOLESCENCE: Adolescence is an exceptional, highly sensitive period when identifications acquired during infancy undergo reorganization. It is a time when defective identification can be reelaborated, favoring structuralization of a "normal" or "pathological" (here maniac depressive) personality depending on the quality of the "identificatory", therapeutic (psychotherapy), or daily life events. PMID- 10686973 TI - [Manic-depressive disorders in adolescence. Lithium treatment]. AB - FIRST INTENTION PREVENTION: Lithium is the first intention preventive treatment for bipolar disorders. It the most effective choice in this indication and provides the most benefit for the patient, particularly the young patient still in the process of maturation. SURVEILLANCE: Serum lithium must be monitored regularly. It is important for the clinician to be aware of different factors which can modify serum levels, including disease states or iatrogenic effects related to co-prescriptions. PATIENT INFORMATION: Long-term compliance and surveillance, and thus treatment efficacy, depend greatly on the quality of the information provided to the patient and his/her family. PMID- 10686974 TI - Inflammatory cytokines and cell response in surgery. AB - The systemic inflammatory response as mediated by the cytokine network is undoubtedly complex. While inflammatory cytokines are indispensable in wound healing and the restoration of homeostasis, it is often the excessive activity of either proinflammatory or anti-inflammatory cytokines that causes injury to the host or renders the host immunocompromised, respectively. Central to the functional biology of cytokines in surgical injury and infections are the responses of immune cells to such insults. It is clear that immunocytes are the source of cytokine production, and these products possess important autocrine, as well as systemic activities. The ability to alter immunocyte function through extracellular hormonal influences or by manipulating intracellular signaling mechanisms are potential strategies for regulating the inflammatory cytokine response during injury. PMID- 10686975 TI - Invited commentary: are the staging systems for lymphatic spread important in gastric cancer? PMID- 10686976 TI - Superiority of a new UICC-TNM staging system for gastric carcinoma. AB - BACKGROUND: The definition of the degree of lymph node metastasis (n classification) for gastric cancer differs greatly in the new Union Internationale Contre le Cancer--TNM classification (5th edition) and the Japanese gastric cancer classification (JGC). The feasibility of the new TNM classification is evaluated in comparison with the JGC. METHODS: At Chiba University, 940 patients who underwent a gastrectomy were retrospectively classified into appropriate stages with both the TNM and JGC systems, and the survival curves of the respective stages were also compared. RESULTS: Patients with 1 to 6 metastatic nodes (TNM-pN1) showed similar survival rates whether the metastases were limited to the perigastric area (JGC-n1) or reached distant areas (JGC-n2). The patients with node metastasis that was limited to the perigastric area (JGC-n1) had significantly different survival rates, depending on the number of metastatic nodes (TNM-pN1 or pN2, P = .022). A similar phenomenon was also observed in patients with TNM-N2 and JGC-n2. A multivariate analysis indicated the TNM N-classification, rather than the JGC n-classification, as an independent prognostic factor. CONCLUSIONS: The new TNM classification appears to be a better prognostic indicator than the JGC system for patients with gastric carcinoma. PMID- 10686977 TI - Gangrenous and perforated appendicitis: a meta-analytic study of 2532 patients indicates that the incision should be closed primarily. AB - BACKGROUND: Surgical incisions after appendectomy for complicated (gangrenous or perforated) acute appendicitis are often managed with delayed closure (DC) rather than primary closure (PC). This study synthesizes the results of other studies in the surgical literature and supports the routine use of PC. METHODS: Studies dealing with complicated appendicitis were reviewed to assess the results of PC in comparison with DC. The rate of incision (wound) infection in groups of patients managed by PC and DC were compared with the use of a statistical technique that defined the probability of expected results by incorporating data derived from all of the various study groups. RESULTS: Of the 2532 patients who had been treated for complicated appendicitis and who were assessed, 1724 patients underwent PC and 808 patients underwent DC. The rate of incision infection was 4.7% and 4.6% in the PC and DC groups, respectively. With a 95% confidence interval, there was no demonstrable difference between the 2 types of operative site management (P < .01). CONCLUSIONS: PC of the skin and subcutaneous tissue after appendectomy for gangrenous or perforated appendicitis, combined with the use of antibiotic therapy in the perioperative period, is not associated with an increased risk of incision infection when compared with DC. PMID- 10686978 TI - Neutrophil-derived serine proteinases enhance membrane type-1 matrix metalloproteinase-dependent tumor cell invasion. AB - BACKGROUND: Matrix metalloproteinase-2 degrades a variety of basement membrane components and is essential for tumor invasion. We have previously reported that membrane type-1 matrix metalloproteinase (MT1-MMP) cooperates with neutrophil derived serine proteinases (NDPs; elastase, cathepsin G, protease-3) to activate matrix metalloproteinase-2. We therefore hypothesized that NDPs enhance tumor cell invasion. METHODS: Clones of human HT1080 fibrosarcoma cells transfected with MT1-MMP sense (HT-SE) or antisense CDNA (HT-AS) were used. These cells express either high (HT-SE) or extremely low levels (HT-AS) of MT1-MMP relative to nontransfected HT1080 cells (HT-WT). The cells were incubated in the presence or absence of purified NDP, with or without alpha 1-antitrypsin or the MMP inhibitor batimastat. Cell invasion was measured with the use of Boyden chambers with polycarbonate membranes coated with a reconstituted extracellular matrix. RESULTS: Under control conditions HT-WT and HT-SE cells were 4-fold more invasive than HT-AS cells. The addition of NDP increased HT-WT and HT-SE cell invasion 60% to 100% but had no effect on HT-AS cells. alpha 1-antitrypsin or batimastat did not decrease the baseline invasiveness of HT-WT and HT-SE cells; however, they abrogated the stimulatory effect of NDP. CONCLUSIONS: HT1080 cell invasion depends on MT1-MMP expression. MT1-MMP overexpression does not increase invasiveness by itself. NDPs increase invasion by MT1-MMP expressing cells by activating matrix metalloproteinase-2. PMID- 10686979 TI - Vascular smooth muscle mechanics in isolated perfused segments of carotid arteries. AB - BACKGROUND: We hypothesized that smooth muscle contraction and relaxation responses in a muscle bath (isometric tension) would be different than responses of intact vessels (isotonic tension). METHODS: Bovine carotid artery contractile responses to the catecholamine, norepinephrine, and smooth muscle relaxant, 3 isobutyl-1-methylxanthine, were examined in strips of vessels in a muscle bath and in intact whole vessels in an isolated perfused whole-vessel perfusion apparatus. RESULTS: The maximal tension in the muscle bath depended on the length of the strip. The responses of whole vessels to increasing pressure was curvilinear. The maximal decrease in vessel diameter in intact vessels in response to the catecholamine and norepinephrine occurred at low intraluminal pressures. The dose-response curve to norepinephrine was shifted to the left in intact vessels compared with strips of vessels in the muscle bath, which suggests that whole vessels were more sensitive to norepinephrine. The maximal increase in diameter to increasing intraluminal pressure occurred in the presence of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, which suggests that there was significant intrinsic tone in the vascular smooth muscle. CONCLUSIONS: These results suggest that there are differences in the contractile properties of the vascular smooth muscle that are related to the ex vivo system used to examine smooth muscle responses. Responses obtained in isolated perfused whole vessels may more closely approximate in vivo responses. PMID- 10686980 TI - Right trisegment portal vein embolization for biliary tract carcinoma: technique and clinical utility. AB - BACKGROUND: Right portal vein embolization has become popular in preparation for right hepatic lobectomy. However, right trisegment portal vein embolization (R3PE) is not well established. METHODS: We performed R3PE in 15 patients with biliary tract carcinoma and 1 patient with primary sclerosing cholangitis. We used 2 types of 5.5 F triple-lumen balloon catheters to embolize portal branches of the right trisegment (the left medial, the right anterior, and the right posterior segments). RESULTS: R3PE was successful in all patients without any complications. The calculated volume of the right lobe significantly (P < .01) decreased from 650 +/- 161 cm3 before embolization to 585 +/- 143 cm3 after embolization; the volume of the left lateral segment significantly (P < .0005) increased from 240 +/- 58 cm3 to 361 +/- 66 cm3. The volume of the left medial segment was unchanged. The volume gain of the left lateral segment was larger in patients with R3PE than in those patients (n = 41) with right portal vein embolization (122 +/- 39 cm3 vs 66 +/- 35 cm3; P < .0001). Two of the 16 patients underwent only laparotomy because of peritoneal dissemination, and the remaining 14 patients underwent right hepatic trisegmentectomy with caudate lobectomy. In addition, portal vein resection was also performed in 5 patients, and pancreatoduodenectomy and right hemicolectomy was performed in 3 patients. One patient died of posthepatectomy liver failure 87 days after surgery, a mortality rate of 7.1% (1/14 patients). CONCLUSIONS: R3PE is more useful than standard right portal vein embolization in preparation for right hepatic trisegmentectomy and has the potential to increase the safety of this high-risk surgery for patients with biliary tract carcinoma. PMID- 10686981 TI - Adenosine prevents activation of transcription factor NF-kappa B and enhances activator protein-1 binding activity in ischemic rat heart. AB - BACKGROUND: Adenosine prevents myocardial TNF-alpha production induced by ischemia/reperfusion, but the mechanisms are poorly understood. Transcription factors NF-kappa B and AP-1 have been implicated in the regulation of a variety of inducible gene expressions in response to oxidative stress and cellular defense. The effects of adenosine on NF-kappa B and AP-1 activation have not been clearly defined. This study demonstrated differential effects of adenosine on NF kappa B and AP-1 nuclear binding activity in ischemic myocardium. METHODS: Isolated working rat hearts were subjected to 0, 1, 2, 3, 4, 5, 7.5, 10, 15, and 30 minutes of ischemia, with 4 to 6 hearts for each time point with and without adenosine (100 mumol/L). NF-kappa B and AP-1 binding activity in the nucleus were analyzed by electrophoretic mobility shift assay (EMSA). I kappa B alpha levels in the cytoplasm were measured by Western blot analysis. TNF-alpha mRNA levels were determined by RT-PCR. RESULTS: NF-kappa B binding activity in the nucleus significantly increased after 4 minutes of ischemia and remained to 30 minutes. The levels of I kappa B alpha protein in the cytoplasm markedly decreased after 4, 5, 7.5, and 10 minutes of ischemia. TNF-alpha mRNA levels peaked after 10 minutes of ischemia. AP-1 DNA binding activity was induced and persisted during all ischemic periods. Adenosine significantly inhibited NK-kappa B binding activity in the nucleus, markedly prevented the loss of I kappa B alpha proteins from the cytoplasm, and concomitantly down-regulated TNF-alpha mRNA expression, but enhanced AP-1 binding activity in the nucleus of ischemic myocardium. CONCLUSIONS: Adenosine modulation of NF-kappa B activation may be the cellular molecular mechanism of down-regulation of TNF-alpha mRNA expression. The cardioprotective properties of adenosine may be involved in the differential modulation of NF-kappa B and AP-1 activation during myocardial ischemia. PMID- 10686982 TI - Myoelectric and motor activity after proctocolectomy and ileal pouch-anal anastomosis in dogs. AB - BACKGROUND: The aim of this study was to investigate the motor function of the ileoanal pouch and to evaluate its coordination with proximal small-intestine motility. METHODS: Proctocolectomy and ileal J pouch-anal anastomosis were performed in 12 dogs. Motility was recorded by serosal electrodes and strain gauge transducers. RESULTS: Transmission of the migrating motor complex (MMC) on the pouch appeared in only 37 of 109 measurements. On the ascending limb there was a constant irregular activity with no MMC detectable. Motility pattern of the pouch did not change postprandially. Spontaneous defecation always appeared independently from MMC transmission without an increase of electrical or mechanical activity or endoluminal pouch pressure. CONCLUSIONS: Ileal pouch motility is independent from motility patterns of the proximal intestine. Its random contractile activity might provide storage function and make the pouch act as a functional reservoir. Intrinsic pouch motility is not responsible for pouch evacuation under physiologic conditions. PMID- 10686983 TI - Conservative surgical treatment of diffuse peritonitis. AB - BACKGROUND: Peritonitis is, even today, a significant source of death and complications. The objective of this study was to determine the morbidity and mortality rates, the incidence of reoperations, and the need for additional treatment strategies (on demand) in patients with diffuse peritonitis. METHODS: Prospective analysis including all patients (n = 258) with diffuse peritonitis admitted to our surgical service between November 1993 and April 1998 who underwent a uniform surgical treatment concept of peritonitis including early intervention, source control, and extensive intraoperative lavage. RESULTS: The 258 patients with diffuse peritonitis averaged a mean Mannheim Peritonitis Index of 27.1 points (range, 11-43 points). Source control at the initial operation was possible in 230 of the patients (89%), of those, 21 patients (9%) needed reintervention. In 28 patients (11%), source control was not possible at the initial operation. Twenty of these patients (71%) had to undergo additional treatment strategies (on demand) such as continuous lavage and/or laparostomy. Overall 228 of the 258 patients (88%) needed just 1 initial surgical intervention. The overall morbidity rate was 41%; the rate of reoperation was 12%, and the hospital mortality rate was 14%. CONCLUSIONS: A conservative surgical treatment concept supplemented with "extensive" intraoperative lavage reduces the reoperation rate compared with other treatment standards of peritonitis and achieves a low mortality rate in patients with diffuse peritonitis. PMID- 10686984 TI - Usefulness of autologous blood transfusion for avoiding allogenic transfusion and infectious complications after esophageal cancer resection. AB - BACKGROUND: A retrospective investigation was conducted to determine whether autologous blood collection could reduce allogenic transfusion after resection of esophageal cancer and whether allogenic transfusion influenced postoperative infection. METHODS: Patients (n = 100) who met the criteria for hemoglobin, age, body weight, and serum protein donated 800 mL of autologous blood from May 1994 to December 1997. The control group (n = 248) was selected from patients who met the same criteria and did not donate autologous blood over the 10 years before the start of autologous blood collection. RESULTS: Only three patients (3%) from the autologous group required allogenic transfusion versus 84 patients (33.7%) from the control group. Sixteen of the 26 patients who received more than 4 units of allogenic blood contracted postoperative infections compared with 25 of 165 patients who did not (P < .0001). Autologous blood transfusion significantly increased the probability of avoiding allogenic transfusion (odds ratio, 27.58), and allogenic transfusion was significantly related to postoperative infection (odds ratio, 1.19), according to logistic regression analysis. CONCLUSIONS: Autologous blood collection reduces the need for allogenic transfusion in patients undergoing resection of esophageal cancer, and avoidance of allogenic transfusion may reduce the risk of postoperative infection. PMID- 10686985 TI - Multiple intrasplenic hepatocyte transplantations in the dalmatian dog. AB - BACKGROUND: Hepatocyte transplantation is an attractive potential treatment for liver-based inborn errors of metabolism and for fulminant hepatic failure. Dalmatian dogs have a metabolic error that results in hyperuricosuria. This report focuses on the effect of multiple, sequential intrasplenic transplants of fresh and cryopreserved hepatocytes in dalmatians. METHODS: Dalmatians underwent intrasplenic hepatocyte transplantation with hepatocytes taken from healthy mongrels. Dalmatian urinary uric acid excretion was measured preoperatively, and this served as the control value. Three hepatocyte transplantations were performed at 30-day intervals--the first with freshly isolated cells, and both the second and the third with cryopreserved hepatocytes from the same donor. Urinary uric acid excretion was measured postoperatively twice per week. RESULTS: The urinary uric acid excretion decreased an average of 54% after the first hepatocyte transplantation. The effect was transient and lasted an average of 22 days (range, 19-50 days). Subsequent intrasplenic hepatocyte transplantation with cryopreserved hepatocytes resulted in similar decreases in urinary uric acid excretion. Each transplant resulted in a significant decrease in urinary uric acid excretion when compared with baseline values (P = < .001). CONCLUSIONS: Sequential intrasplenic hepatocyte transplantation is feasible in this model. This method provided a significant, but transient, correction in urinary uric acid excretion that was similar with either fresh or cryopreserved hepatocytes. A substantial biologic effect provided by cryopreserved hepatocytes has important implications in clinical hepatocyte transplantation. PMID- 10686986 TI - Esophagogastric junction pressure topography after fundoplication. AB - OBJECTIVES: This study compared the pressure topography after laparoscopic Nissen fundoplication to that of normal subjects and patients with hiatal hernia and reflux disease. METHODS: Seven patients with fundoplication, 7 normal subjects and 7 patients with hiatal hernia, were studied. The squamocolumnar junction and intragastric margin of the esophagogastric junction (EGJ) were marked with metal clips. Axial and radial characteristics of EGJ pressure were mapped relative to the hernia and clipped during concurrent fluoroscopy and manometry. Responses to inspiration and abdominal compression were also analyzed. RESULTS: Fundoplication modifies the EGJ by restoration of the hiatal component of EGJ pressure and elongation of the subdiaphragmatic component. Maximal EGJ pressure after fundoplication is mainly dependent on the extrinsic effect of the hiatal canal that compresses the esophagus; the resultant length of the EGJ reflects the length of the fundic wrap. Integrity of the EGJ after fundoplication is independent of the intrinsic lower esophageal sphincter itself. CONCLUSIONS: Fundoplication alters the pressure topography of the EGJ by reducing the hiatal hernia, tightening the hiatal orifice, and constructing a subdiaphragmatic wrap of variable length. Each effect depends on different technical aspects of the surgery with the potential of substantial variability in the resultant pressure topography. PMID- 10686987 TI - Neutrophils express tissue factor in a monkey model of sepsis. AB - BACKGROUND: Although tissue factor (TF) is involved in hemostasis, thrombogenesis, inflammation, and cellular immune response, its source in sepsis remains controversial. Recently, we found that, in addition to monocytes and endothelial cells, neutrophils may express TF in a rabbit model. The purpose of this study was to determine whether neutrophils could be a source of TF in a monkey model of sepsis. METHODS: TF messenger RNA (mRNA) and protein in neutrophils were assayed by in situ hybridization and immunohistochemistry in tissues obtained from monkeys after injection of lipopolysaccharide (LPS) (n = 3) and after injection of saline as a control (n = 2). Coagulation parameters were measured before and at 1.5 and 3 hours after injections. RESULTS: In LPS-treated monkeys, TF mRNA and protein were induced not only in monocytes and endothelial cells, but also in neutrophils accumulating in the liver 3 hours after LPS injection. Thrombin-antithrombin III complex and fibrin degradation products D dimer levels were significantly increased at 3 and 1.5 hours after LPS injection compared with controls. CONCLUSIONS: Neutrophils are a source of TF and are implicated in direct activation of the coagulation cascade in the early phases of sepsis in the monkey. These results give important information for the treatment of sepsis. PMID- 10686988 TI - A prospective comparison of systemic-bladder versus portal-enteric drainage in vascularized pancreas transplantation. AB - BACKGROUND: Most pancreas transplants are performed with systemic venous delivery of insulin and bladder drainage of the exocrine secretions (systemic-bladder [S B]). To develop a more physiologic procedure, we performed pancreas transplantations with portal venous delivery of insulin and enteric drainage of the exocrine secretions (portal-enteric [P-E]). METHODS: During an 11-month period, we prospectively alternated 32 consecutive pancreas transplant recipients to either S-B (n = 16) or P-E (n = 16) drainage with standardized immunosuppression. RESULTS: Patient, kidney, and pancreas graft survival rates after simultaneous kidney-pancreas transplantation were 91% S-B versus 92% P-E, 91% S-B versus 92% P-E, and 82% S-B versus 92% P-E, respectively. Pancreas graft survival rates after solitary pancreas transplantation were 80% S-B versus 75% P E. There were no graft losses either to immunologic or infectious complications in either group, but the incidence of acute rejection was slightly higher in the S-B group (44% S-B vs 31% P-E, P = NS). The cost and length of the initial hospital stay were similar between groups. The incidence of operative complications, major infections, and cytomegalovirus infections were likewise comparable. However, the S-B group was characterized by a slight increase in the number of readmissions, urinary tract infections, and urologic complications. Furthermore, metabolic acidosis and dehydration were more common in the S-B group. CONCLUSIONS: Pancreas transplantation with P-E drainage can be performed with short-term results comparable to those of transplantation with S-B drainage. PMID- 10686989 TI - A surgeon is an amalgam of many elements. PMID- 10686990 TI - Successful transplantation in a patient with ruptured large hepatocellular carcinoma with diaphragmatic invasion. PMID- 10686991 TI - The use of intra-aortic counterpulsation balloon for the treatment of cerebral vasospasm and edema. PMID- 10686992 TI - Surgical management of inflammatory abdominal aortic aneurysm associated with occult aortocaval fistula. PMID- 10686993 TI - Malignant colorectal cell spread during no-touch cancer surgery. PMID- 10686994 TI - Misclassification of hospital procedure volume in surgical outcomes studies. PMID- 10686995 TI - USDA to help pets in distress. PMID- 10686996 TI - Treatment advances reported at equine meeting. PMID- 10686997 TI - Meat irradiation gets green light, approval process streamlined. PMID- 10686998 TI - Concerning wild canids and wolf hybrids. PMID- 10686999 TI - Requests for cooperation between private practitioners and shelter veterinarians. PMID- 10687000 TI - In support of veterinary pathologists. PMID- 10687001 TI - What is your diagnosis? Tracheal collapse at the level of the sixth cervical vertebra. PMID- 10687002 TI - Theriogenology question of the month. Specific aversion to handling for semen collection and to personal approaching the genital area. PMID- 10687003 TI - Genetic counseling for cat and dog owners and breeders--managing the emotional impact. PMID- 10687004 TI - Results of a survey on educational and research programs in complementary and alternative veterinary medicine at veterinary medical schools in the United States. AB - OBJECTIVE: To document educational and research programs in complementary and alternative veterinary medicine (CAVM) at US veterinary schools and to develop recommendations for additional curriculum development and research in these modalities. DESIGN: Mail questionnaire. SAMPLE POPULATION: Deans, curriculum committees, and interested faculty at US veterinary schools. PROCEDURES: Questionnaires were mailed to personnel at all 27 US veterinary schools. Nonrespondents received a follow-up letter and telephone contact. Information was used to establish the current status of CAVM. RESULTS: Responses were received for 41 of 120 (34%) questionnaires. Responses were received from 23 of 27 veterinary schools, but number of respondents varied at each institution (range, 1 to 4) and some surveys were not complete. Seven of 27 US veterinary schools had an educational program in CAVM. Thirty-six (87%) respondents believed that acupuncture, nutraceuticals, nutritional supplements, and physical therapy should be included in the curriculum, 25 (61%) indicated that botanical (herbal) medicine should be included, and 25 (61%) believed that chiropractic should be included. Only 17 (44%) respondents believed that homeopathy should be included. The majority of respondents believed that CAVM should be offered as elective courses. Research in CAVM has been conducted at 6 responding schools. CONCLUSIONS: Currently, few veterinary schools offer educational or research programs in CAVM. Veterinary schools are aware of the interest in CAVM and acknowledge a lack of educational and research programs in these areas. More veterinary schools are in the process of developing educational and research programs in various aspects of CAVM. PMID- 10687005 TI - Types and doses of injectable medications given to periparturient sows. AB - OBJECTIVE: To determine types and doses of injectable medications given to periparturient sows and reasons for administering those medications, and to compare medication practices among farms of different sizes. DESIGN: Survey. SAMPLE POPULATION: 301 farms; 231,016 periparturient sows. PROCEDURE: A survey was used to obtain information regarding medications given to sows during the farrowing period. State and federal veterinary medical officers completed surveys during their final interview with producers who had participated in the National Animal Health Monitoring System's (NAHMS) Swine 95 study. Data were summarized and treatment regimens compared among farms of different sizes. RESULTS: More than a third of the sows received medications during the farrowing period. The most common reasons for administering medications were routine preventive treatment and treatment of dystocia, uterine discharge, and poor appetite. The most commonly used medications for treatment of sick sows were oxytocin, procaine penicillin G, and B vitamins. A high percentage of medications were either not indicated for the specific condition or used at greater or less than the approved dose. In general, treatment rates and medications used did not differ among farms of different sizes. CONCLUSIONS AND CLINICAL RELEVANCE: Better treatment protocols are needed to provide more appropriate treatment of sick sows. PMID- 10687006 TI - Peliosis hepatis in a dog infected with Bartonella henselae. AB - A 6-year-old spayed female Golden Retriever was examined because of generalized weakness and abdominal distention. Abdominal ultrasonography revealed a large quantity of peritoneal fluid. In addition, the liver appeared larger than normal and contained multiple, small, nodular masses and cyst-like structures. Abdominal exploratory surgery was performed, and 5 L of serosanguineous peritoneal fluid was removed. Gross lesions were not found in the stomach, kidneys, intestines, adrenal glands, or urinary bladder. There were diffuse cystic nodules in all liver lobes. The dog did not recover from anesthesia. A diagnosis of peliosis hepatis was made on the basis of gross and histologic appearance of the liver. A polymerase chain reaction assay revealed Bartonella henselae DNA in liver specimens. To our knowledge, this is the first report of molecular evidence of B henselae infection in a dog with peliosis hepatis. PMID- 10687007 TI - Epitheliotropic T-cell gastrointestinal tract lymphosarcoma with metastases to lung and skeletal muscle in a cat. AB - An Abyssinian cat examined because of hypoproteinemia and weight loss was found to have epitheliotropic T-cell lymphosarcoma of the gastrointestinal tract. Neoplastic lymphocytes infiltrated the epithelium and lamina propria of the stomach, small intestine, and large intestine. Metastatic foci were found in the lungs and the musculature of the right hind limb. Chemotherapy resulted in transient shrinkage of the lung and limb masses; however, these masses grew to approximately their original size within 2 weeks after initiation of treatment, and the cat was euthanatized. PMID- 10687008 TI - Clinical, radiographic, and pathologic features of bronchiectasis in cats: 12 cases (1987-1999). AB - OBJECTIVE: To determine clinical, radiographic, and pathologic features of bronchiectasis in cats. DESIGN: Retrospective study. ANIMALS: 12 cats with histologic evidence of bronchiectasis. PROCEDURE: Information on signalment, body weight, clinical signs of respiratory disease, concurrent diseases, method by which lung tissue specimens were obtained (surgical biopsy or necropsy), and histopathologic findings was obtained by reviewing medical records from January 1987 to June 1999 for cats with confirmation of bronchiectasis by histologic examination. Available thoracic radiographs were reviewed by a board-certified radiologist. RESULTS: Bronchiectasis was most commonly identified in older male cats. Clinical signs referable to the lower portion of the respiratory tract were detected in only 5 cats but, when evident, were usually chronic (duration > 1 year). Radiographic pattern of bronchiectasis was cylindrical in 4 cats, and in 1 of these cats, a saccular pattern was also identified. For most cats, bronchiectasis was detected in a single lung lobe. Radiographic evidence of bronchiectasis was not detected in 2 cats. Concurrent respiratory diseases included chronic bronchitis and bronchiolitis, neoplasia, bronchopneumonia, endogenous lipid pneumonia, and emphysema. CONCLUSIONS AND CLINICAL RELEVANCE: Bronchiectasis appears to be an uncommon respiratory tract disorder that predominantly affects older male cats. Thoracic radiography may not be sensitive for the diagnosis of bronchiectasis in cats. Bronchiectasis in cats appears to be a sequela of chronic inflammatory bronchopulmonary diseases, especially chronic bronchitis, neoplasia, and bronchopneumonia. PMID- 10687009 TI - Descriptive epidemiologic study of disease associated with influenza virus infections during three epidemics in horses. AB - OBJECTIVE: To describe 3 epidemics of respiratory tract disease caused by influenza virus infections in a large population of horses. DESIGN: Cross sectional and prospective longitudinal observational studies. ANIMALS: All horses stabled at a Thoroughbred racetrack. PROCEDURES: During a 3-year period, descriptive information was collected as horses arrived at the racetrack and throughout race meetings. Routine observations and physical examinations were used to classify horses' disease status. Cause of epidemics was established by use of serologic testing and identification of influenza virus in nasal secretions. RESULTS: An epidemic of respiratory tract disease caused by influenza virus infections was identified during each year of the study. Attack rates of infectious upper respiratory tract disease (IURD) ranged from 16 to 28%. Incidence of disease caused by influenza virus infections during racing seasons in the second and third years was 27 and 37 cases/1,000 horses/mo, respectively. Physical distributions of stall locations revealed that affected horses were stabled throughout the population; horses affected later in epidemics were often clustered around horses affected earlier. Mucopurulent nasal discharge and coughing were observed in 83 and 62% of horses with IURD, respectively. Median duration of clinical disease was 11 days. Serologic testing was the most sensitive method used to detect influenza virus infections; 76% of affected horses seroconverted to influenza virus. CONCLUSIONS AND CLINICAL RELEVANCE: Epidemics of IURD were observed annually in association with influenza virus infections. Few precautions were taken to limit spread of infection. Preventing or decreasing the likelihood of exposure and improving immunity in the population could substantially decrease risk of disease in similar populations. PMID- 10687010 TI - Risk factors for disease associated with influenza virus infections during three epidemics in horses. AB - OBJECTIVE: To identify risk factors associated with respiratory tract disease in horses during 3 epidemics caused by influenza virus infections. DESIGN: Cross sectional and prospective longitudinal observational studies. ANIMALS: 1,163 horses stabled at a Thoroughbred racetrack. PROCEDURES: Investigations were conducted during a 3-year period. An epidemic of respiratory tract disease caused by influenza virus infections was identified in each year. Routine observations and physical examinations were used to classify horses' disease status. Data were analyzed to identify factors associated with development of disease. RESULTS: Results were quite similar among the epidemics. Concentrations of serum antibodies against influenza virus and age were strongly associated with risk of disease; young horses and those with low antibody concentrations had the highest risk of disease. Calculation of population attributable fractions suggested that respiratory tract disease would have been prevented in 25% of affected horses of all horses had high serum antibody concentrations prior to exposure. However, recent history of vaccination was not associated with reduction in disease risk. Exercise ponies had greater risk of disease than racehorses, which was likely attributable to frequent horse-to-horse contact. CONCLUSIONS AND CLINICAL RELEVANCE: Particular attention should be paid to young horses, those with low serum antibody concentrations, and horses that have frequent contact with other horses when designing and implementing control programs for respiratory tract disease caused by influenza virus infections. It appears that control programs should not rely on the efficacy of commercial vaccines to substantially reduce the risk of disease caused by influenza virus infections. PMID- 10687011 TI - Squamous cell carcinoma of the urethral process in a horse with hemospermia and self-mutilation behavior. AB - A 14-year-old Arabian stallion was examined because of acute hemospermia. The stallion was used in an artificial breeding program and had a 6-year history of low-grade hemospermia and a 4-year history of self-mutilation behavior. During previous examinations, minor irritation of the urethral process was identified as the source of the bleeding. Physical examination revealed a mucosal ulceration in the distal portion of the urethra. Histologic examination of a biopsy specimen from this area revealed low-grade squamous cell carcinoma. The urethral process was excised, and the hemospermia resolved. Frequency of self-mutilation behaviors also decreased after surgery, suggesting that there may have been a link between irritation of the urethral process and development of self-mutilation behavior. PMID- 10687012 TI - Causes of poor performance of horses during training, racing, or showing: 348 cases (1992-1996). AB - OBJECTIVE: To determine results for horses undergoing a high-speed treadmill examination, including videoendoscopy of the pharynx and larynx before and during exercise, echocardiography before and after exercise, and electrocardiography before, during, and after exercise, because of poor performance. DESIGN: Retrospective study. ANIMALS: 348 horses. RESULTS: A definitive diagnosis was obtained for 256 (73.5%) horses. One hundred forty-eight horses had dynamic obstruction of the airway during exercise, 33 had clinically important cardiac arrhythmias alone, 22 had a combination of dynamic airway obstruction and clinically important cardiac arrhythmias, 19 had poor cardiac fractional shortening immediately after exercise, 10 had exertional rhabdomyolyis, 15 had clinically apparent lameness, and 9 had other disorders. Thirty-nine of the horses with dynamic obstruction of the airway during exercise had multiple airway abnormalities. Fifty-three horses also had subclinical myopathy CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a complete evaluation, including a high speed treadmill examination, should be conducted in horses with poor performance, regardless or whether horses do or do not have a history of abnormal respiratory noises and particularly if the horses have grade-II or -III left laryngeal hemiplegia. PMID- 10687013 TI - Evaluation of assays for determination of passive transfer status in neonatal llamas and alpacas. AB - OBJECTIVE: To evaluate several practice-adapted assays for determination of passive transfer status in crias. ANIMALS: 24 llama and 9 alpaca crias. DESIGN: Prospective study. PROCEDURE: Serum IgG concentration was measured by use of a radial immunodiffusion assay when crias were 45 to 51 hours old. Results were compared with serum gamma-glutamyltransferase (GGT) activity, serum total protein, albumin, globulin, and total solids concentrations, and results of commercially available and traditional sodium sulfite turbidity (SST) tests. RESULTS: Mean (+/- SD) serum IgG concentration was 1,762 +/- 1,153 mg/dl. On the basis of a threshold value of 1,000 mg of IgG/dl at 48 hours of age, 5 of 33 (15.15%) crias had failure of passive transfer. Serum total solids, protein, and globulin concentrations were significantly associated with serum IgG concentration, whereas serum GGT activity and serum albumin concentration were not. Serum IgG concentrations were significantly different among crias with negative, 2+, and 3+ scores on the traditional SST test. Serum IgG concentrations were not significantly different between crias with negative and 100 mg/dl scores or 100 and 300 mg/dl scores on the commercially available SST test. However, all other comparisons between crias with different scores revealed significant differences. Sensitivity and specificity ranged between 0 and 1, depending on the test and endpoint selected. CONCLUSIONS AND CLINICAL RELEVANCE: The commercially available SST test and determination of serum total protein and globulin concentrations are suitable methods for assessing passive transfer status in llama and alpaca crias. PMID- 10687014 TI - Diagnosis and surgical repair of patellar luxations in a flock of sheep. AB - Eleven sheep with a common ancestry were evaluated for clinical signs of hind limb lameness. Physical examination revealed advanced forms of patellar luxation. Radiography confirmed the diagnosis of patellar luxation; flexed dorsoproximal dorsodistal radiographic views were obtained and revealed hypoplasia of the femoral trochlea. Because of the advanced form of patellar luxation and trochlear hypoplasia, surgical intervention was recommended in the form of recession trochleoplasty and a modified tibial tuberosity transposition, along with imbrication and releasing incisions. Sheep may have a genetic predisposition for patellar luxation. PMID- 10687015 TI - [Three patients with unrecognized orthostatic intolerance]. AB - Three patients, men aged 20, 50 and 56 suffered from orthostatic intolerance and syncope for years, despite visits to many doctors. Medical history and cardiovascular reflex investigation revealed problems with the orthostatic blood pressure regulation, caused by initial orthostatic dysregulation, vasovagal syncope and pure autonomic failure, respectively. A structured medical history as well as blood pressure measurements in the supine and standing positions can reveal the diagnosis in many instances of syncope. After confirming the diagnosis, the first concern is a clear explanation of the underlying problems to the patient. Treatment with volume expansion should be considered. PMID- 10687016 TI - [Handling small relative risks in science and management: the third-generation pill]. AB - Small relative risks (a twofold or lesser increase in disease frequency) become scientifically acceptable with (a) repeated consistent findings from studies that (b) address the most important forms of bias and confounding, and (c) when there is a plausible biologic mechanism. The third-generation oral contraceptive controversy is an example of such a relevant but small relative risk and demonstrates the problem of interpretation and implementation into medical practice guidelines. PMID- 10687017 TI - [Coagulation disorders in cancer patients: possible opportunity for therapy]. AB - Coagulation disorders occur often in cancer patients. Thrombosis or embolism may be the first sign of an underlying malignancy. In addition, subclinical coagulation disturbances have been found in the blood of cancer patients, for example elevated concentrations of tissue factor or thrombin-antithrombin complexes. In 20% of the patients thrombocytosis occurs, and for lung and colon cancer it was found that thrombocytosis is an independent negative prognostic factor for survival. The role of an activated coagulation cascade in tumour growth is not completely clear, but there is strong evidence that the formation of a temporary fibrin matrix stimulates the formation of new blood vessels (angiogenesis) and supports tumour growth and metastasis formation. Preclinical investigations demonstrated that tumour growth and metastasis formation can be inhibited by anticoagulants. Clinical studies suggest a beneficial effect of anticoagulants on the survival of cancer patients, but phase III randomised clinical trials should be performed to determine the effect of longterm administration of anticoagulants. PMID- 10687018 TI - [Gastro-intestinal surgery and gastroenterology. V. Chronic pancreatitis: gastroenterologic aspects]. AB - Chronic pancreatitis causes irreversible damage to the pancreatic parenchyma and ultimately leads to exo- and endocrine insufficiency. In the Western world, alcohol is the main cause of chronic pancreatitis; part of the patients with idiopathic or alcoholic pancreatitis conceivably have a raised sensitivity to the toxic effects of alcohol because of a certain genetic predisposition. The most striking symptom, severe recurrent or persistent pain is often difficult to manage. Various forms of imaging examination provide complementary information on lesions of the pancreatic parenchyma, the pancreatic duct, the bile ducts and adjacent structures; ultrasonoscopy is the technique of first choice in case of suspicion of a pancreatic disorder. Endoscopic therapies are booming: sphincterotomy, calculus extraction, insertion of an endoprosthesis in the pancreatic duct and drainage of pseudocysts (transpapillary or through the jejunum or stomach). PMID- 10687019 TI - [Gastrointestinal surgery and gastroenterology. VI. Chronic pancreatitis: surgical aspects]. AB - The indications for surgical treatment of chronic pancreatitis are intractable pain or local complications. The purpose of preoperative investigation, apart from establishing the indications for operation, is to select the kind of procedure to be performed. Important factors include narrowing or dilatation of the pancreatic duct and the presence or absence of an inflammatory mass in the pancreas and of pseudocysts. A pylorus preserving pancreatoduodenectomy or duodenum preserving pancreatic head resection is performed in case of an inflammatory mass in the pancreatic head. In limited clinical trials, duodenum preserving resection was associated with better recovery, this also applies to the Frey procedure, consisting of local resection of the pancreatic head in combination with lateral pancreaticojejunostomy. In case of dilatation of the pancreatic duct (> 8 mm), without an inflammatory mass drainage of the pancreatic ductal system by a lateral pancreaticojejunostomy is appropriate. A pancreatic tail or body resection can be performed for inflammatory lesions confined to the pancreatic tail or body. Symptomatic pseudocysts are drained internally into the stomach, duodenum or a jejunal loop. PMID- 10687020 TI - [Limited changes in prescription of third-generation oral contraceptives after reports on elevated thrombosis risk and subsequent professional guidelines]. AB - OBJECTIVE: To compare the prescription behaviour of general practitioners (GP) and gynaecologists regarding 2nd and 3rd generation oral contraceptives (OC) in the period before and after the warning of increased thrombosis risk of 3rd generation pills. DESIGN: Retrospective, descriptive. METHOD: With the aid of data on OC prescriptions collected by the Institute for Medical Statistics, the numbers of prescriptions from Dutch GPs and gynaecologists in the year before the warning by the British Committee on Safety of Medicines that 3rd-generation OC increased the risk of thrombosis, were compared with the numbers in the year after publication of the guideline of the Dutch General Practitioners Society (NHG), which stated a preference for the 2nd-generation OC, and of the guideline of the Dutch Society for Gynaecology and Obstetrics, which stated no preference. RESULTS: In neither the GP nor the gynaecologist groups did the proportion of 3rd generation pills in the OC prescriptions decrease significantly. However, in the GP group the proportion of new 3rd generation OC prescriptions decreased significantly from 56 to 40%. However, after a few trimesters it rose again. CONCLUSION: The changes in prescription behaviour of the GP group were in accordance with the NHG guidelines. However, the proportion of the 3rd-generation OC prescriptions remained relatively large and the effect was of short duration. The only conclusion that could be drawn regarding the gynaecologists, was that the proportion of 3rd-generation OC prescriptions did not change significantly. PMID- 10687021 TI - [The effect of oral contraception on society]. AB - Oral contraception was introduced 40 years ago. In the Netherlands 'the pill' was more widely accepted than in most other countries. Oral contraception turned family planning into a medical responsibility, which made it respectable. It also separated sexuality from reproduction, which had rather far-reaching consequences for sexual behaviour and experience, as well as for sexual morality. The most important immediate impact of the pill was a sudden acceleration of the downward trend of the birth rate, and a substantial reduction in the number of unwanted pregnancies and so-called forced marriages. Its impact on world population growth has been less marked, mainly because of its costs, as on a global level sterilization and intrauterine devices are more important methods of contraception. The number of abortions in the Netherlands (5-6.5/1000 fertile women/year) has not decreased, but the current possibilities for family planning have led to a change in motivation from 'utterly unwanted pregnancy' to 'unintended pregnancy'. PMID- 10687022 TI - [Favorable effects of vitamin A in measles infection]. AB - The recent measles epidemic that hit the Netherlands in 1999 resulted in numerous hospitalisations and several fatalities, and resembled the previous epidemic of 1987-1988 in numbers and severity. The triple (parotitis epidemica, measles, and rubella) vaccine used in the nationwide, free-of-charge immunization programme is highly effective, but is not accepted for ideological reasons by specific groups in the Dutch community. High oral doses of vitamin A have been shown to reduce mortality and pulmonary and gastrointestinal complications of measles in children in developing countries, but this treatment option is little known to physicians in the Netherlands. The appropriate dose regimens for safe administration of vitamin A in complicated measles are: age under 6 months 50,000 IU, age between 6 months and 2 years 100,000 IU, and age over 2 years 200,000 IU, administered by mouth upon admission. A repeated dose can be administered on the following day. In the Netherlands, and elsewhere, universal measles immunisation remains the first goal in the fight against this highly contagious disease. PMID- 10687023 TI - The Child Behavior Checklist (CBCL) and related material: standardization and validation in Danish population based and clinically based samples. AB - The Child Behavior Checklist (CBCL) and related material, developed by Achenbach and Edelbrock in Vermont, was validated in a mailed survey. A population based sample of 779 children between the age of four and 17 years was compared to a sample of 146 children referred for child psychiatric service. Danish children scored very much like our Scandinavian and German neighbours, but low compared to most others. The CBCL mean 'total behavior problem score' in the population was 17.7. The checklists, especially the parent and teacher versions, provided good construct validity. Youths generally reported more emotional problem behavior than their parents and teachers did about them. In general, parents and youths agree more, reporting emotional problems, and parents and teachers agree more, when scattering externalizing behavior. Short screening constructs are introduced, and by the use of latent trait analysis, four clinically relevant sub scales were generated. Predictive value, sensitivity, specificity and clinical validity must be undertaken in a future two-phase study. PMID- 10687024 TI - Has the Department of Health backed itself into a corner?: Anonymization of data may not protect against breach of confidentiality. PMID- 10687025 TI - Drug-induced hepatotoxicity: pharmacokinetic perspectives and strategies for risk reduction. PMID- 10687026 TI - Free radicals as mediators of alcohol toxicity. AB - In this article we have reviewed recent evidence in support of the hypothesis that acute/chronic alcohol toxicity is mediated primarily via the generation of damaging free radical species in various tissues. Studies in man, animal model or in vitro experimental systems have shown: (1) the demonstration of alcohol induced free radical species directly via esr spectroscopic analysis; (2) increases in indirect markers of ethanol-induced free radical damage in tissues, such as lipid peroxides and protein carbonyl; (3) ethanol-induced alterations in the levels of endogenous tissue antioxidants. These data show the induction of free radicals by ethanol to be a complex interactive process. The classical pathway for ethanol metabolism, catalysed by alcohol dehydrogenase to form acetaldehyde, results in the formation of free radicals, resulting from concomitant changes in NADH levels and NADH/NAD+ redox ratios, which in turn modulate the activity of the free radical generating enzyme xanthine oxidase. The induction of CYP 2E1 in the microsomes results in the generation of HER, another major route by which ethanol induces free radical formation. In addition to the above, ethanol may also induce free radical formation via the reaction of aldehyde oxidase with acetaldehyde or NADH to generate oxyradicals via disturbance in the metabolism of the pro-oxidant iron, or via increased efflux from mitochondria following altered mitochondrial oxidative metabolism. PMID- 10687027 TI - [Antimicrobial agents in the future. The contribution of genomics to their design]. PMID- 10687028 TI - [The outlook for the design of preparations active in relation to resistant forms of bacteria]. PMID- 10687029 TI - [The origin, evolution and clinical significance of antibiotic resistance]. PMID- 10687030 TI - [The importance of fluoroquinolones in treating pneumonia in the elderly]. PMID- 10687031 TI - [The place of fluoroquinolones in the treatment of bacterial infections]. PMID- 10687032 TI - [Antibiotics and the macroorganism]. PMID- 10687033 TI - [Arterial hypoxemia and liver disease: a challenge for pneumology in the next millennium]. PMID- 10687034 TI - [Descriptive analysis (clinical and functional characteristics) of an asthmatic population in a health care district]. AB - OBJECTIVE: To determine the social, demographic, clinical and lung function characteristics of the population of asthmatics living in Health Care District 2 of Madrid. STUDY DESIGN: Transversal (initial visit) phase of a primary care community intervention study. The target population consisted of all asthmatics in District 2 of Madrid who were registered at the public health clinics, were older than 14 and had experienced asthma symptoms within the past year. The selection of patients for the study was systematic (not probabilistic), such that all registered asthmatics seeking care and/or scheduled for check-ups were enrolled. Participation was 96.6% of those on file. We recorded social, demographic, clinical, and lung function variables as well as results of allergy studies, family histories of asthma and/or atopy and type of treatment. RESULTS: Six hundred fifty patients were enrolled, 238 men (36.7%) and 412 women (63.3%). Mean age was 45 +/- 28.1 yr, mean FEV1% was 87.2 +/- 26%. Mean course of disease was 16 +/- 14.6 yr and age of onset was 26 +/- 20 yr. Family history of asthma was present in 34% of cases and a family history of atopy was reported by 21%. Half had allergic rhinitis as an associated factor, with respiratory infections (62.4%) being the most common trigger, followed by pollen (25.3%) and exposure to tobacco smoke (20%). Active smokers accounted for 9.1% of the group, and ex smokers for 21.6%. Short-term beta-adrenergic drugs on demand were used by 64% of patients, whereas 83.2% reported daily and long-term use of beta-2 drugs and 77% used inhaled steroids. Severity of disease differed significantly by age of patient, age at onset and predominance of perennial asthma (with older age and age of onset and greater seasonality observed among severely asthmatic patients). CONCLUSIONS: a) Most symptomatic asthmatic have mild disease. b) Rationally adjusted, appropriate drug treatment should be graded according to level of disease severity. PMID- 10687035 TI - [Usefulness of fiberoptic bronchoscopy in critical pediatric care]. AB - Fiberoptic bronchoscopy (FB) is being applied increasingly in pediatrics as a therapeutic and diagnostic technique. OBJECTIVE: To analyze the contribution of FB to the diagnosis and treatment of respiratory disease in patients admitted to the pediatric intensive care unit (PICU). PATIENTS AND METHOD: We reviewed FB performed with 3.5 and 2.2 mm external diameter instruments between January 1989 and October 1998 in patients admitted to the PICU. Underlying disease and purpose of and indications for FB were analyzed. We also analyzed route of insertion, findings of airway inspection and bronco-alveolar lavage (BAL), complications and the contribution of FB to patient management. RESULTS: A total of 51 procedures in 47 patients aged between ten days and 12 years old were performed. Twenty-one children (41%) were under one year old. The initial indications for FB were diagnostic in 73% and therapeutic in 27%. Airway inspection showed abnormality in 65%. BAL was performed in 18 cases, with microbiological findings in 8 of the 18. The patients benefited directly from the technique in 75% of cases. CONCLUSIONS: FB is useful diagnostic and therapeutic procedure in PICU patients and is generally well tolerated. PMID- 10687036 TI - [Asthma emergencies: can we lower the rate of readmission after discharge?]. AB - INTRODUCTION: The rate of readmission among asthmatic emergency patients varies. In 1991 we observed a 9% rate of readmission following emergency room release. Studies of the number of readmissions or request for medical care are used as the basis for recommendations for releasing patients from hospital emergency care. No studies have assessed disease stability following release or factors related to stability. OBJECTIVES: To assess the course of disease and clinical stability of patients in the period immediately following release from emergency room care. To determine factors that might predict such stability. to determine the rate of readmission in the month following release after applying a treatment protocol and release criteria, with follow-up examination 72 h later. MATERIAL AND METHODS: Prospective, descriptive study with follow-up 72 h and one month after release. SETTING: Emergency and pneumology departments of a general hospital. PERIOD: six months. PATIENTS: 82 asthmatic patients released from the emergency room. RESULTS: Two patients (2.43% were readmitted. At the first follow-up visit (72 h) 81 patients (98.78%) were seen. At the second visit, 66 patients (80.5%) were examined. We observed stability in 70.3% of patients at 72 h and in 86.4% after on month. Stability was statistically related to whether peak expiratory flow greater or less than 70% (76.92% stable versus 46.66% unstable) (p < 0.05). No other clinical, epidemiological or treatment variables recorded upon release were found to influence stability. CONCLUSIONS: 1) A large proportion of patients are in stable condition 72 h after release. 2) When peak expiratory flow upon release is > 70%, stability is significantly increased 72 h later. 3) Our 2.43% rate of readmission one month after release is very low. 4) No differences in stability were seen to be related to oral corticoid prescription upon release. PMID- 10687037 TI - [Nasal nicotine spray in smoking cessation. Results of a multicenter study]. AB - We have carried out an open multicenter follow-up study of the efficacy of a smoking cessation therapy that combined psychological support with use of a nicotine nasal spray. Fifty-seven subjects (37 men, 20 women) with a mean age of 40.3 +/- 15.7 yr and smoking 37.4 +/- 4.7 cigarettes per day were enrolled. The mean Fagerstrom test score was 8.9 +/- 1.1. Patients received minimal psychological support and were prescribed a nicotine nasal spray at the recommended dose of 1 to mg/h for use while awake for a period of three months, with gradual reduction of dose. Subjects were seen on six occasions (on the first day of consultation; 1, 2 and 6 weeks after quitting; 3 and 6 months after quitting). After three months of follow-up, 22 patients (39%) were abstinent; six months after first trying to quit, only 20 of the 57 enrolled had succeeded (35%). Although most subjects (over 90% in the first 15 days, and over 50% at three months) used the treatment, only a small percentage (3%) followed the appropriate doses in the first 15 days and 31% reported doing so at the three month check-up. The mean score reflecting withdrawal syndrome tripled over baseline level during the first six weeks of follow-up. Over three quarters of the subjects suffered side effects caused by the spray, the most common being nasal irritation, rhinorrhea and tearing. Five patients (87%) lef the study because of intolerance to medication. In conclusion, our study found a rate of success of 35% after six months of follow-up. Use of the prescribed medication was inadequate; withdrawal syndrome was more intense and the prevalence of side effects increased during the early treatment period. PMID- 10687038 TI - [A comparative study of patients with chronic obstructive pulmonary disease with and without obstructive sleep apnea syndrome]. AB - We aimed to study whether the presence of obstructive sleep apnea syndrome (OSAS) in patients with chronic obstructive pulmonary disease (COPD) led to differences in clinical picture, gas exchange during awake and sleep states and mechanical ventilation, in comparison with patients with COPD alone. We enrolled 48 COPD patients. In 26 (54.1%), OSAS was ruled out (non-OSAS COPD group) by polysomnography, and in 22 (45.8%) associated OSAS was diagnosed (OSAS COPD group). Patients in the OSAS COPD group experienced greater daytime sleepiness and less dyspnea. Body mass index was not significantly difference. The OSAS COPD group had significantly lower daytime PaO2 (66.4 +/- 10.4 mmHg in the OSAS COPD group and 75.5 +/- 11.2 mmHg in the non-OSAS COPD group; p = 0.01); there were no differences in PaCO2.Pimax in the OSAS-COPD group was 70.6 +/- 23.8 cmH2O, a level that was significantly lower than in the non-OSAS COPD group (Pimax 90.5 +/ 26.1 cmH2O; p = 0.04). Patients in the non-OSAS COPD group experienced longer periods of REM sleep. Nighttime saturation parameters were significantly different in the group with OSAS. We conclude that patients with both OSAS and COPD experience greater oximetric changes than those without OSAS, during both sleep and awake states. The deterioration of respiratory muscle pressures in such patients may play an important role in the changes. The groups also present differences in the intensity of some symptoms, such as degree of daytime sleepiness and dyspnea. PMID- 10687039 TI - [The management trap in medicine. Introduction to a needed discussion]. PMID- 10687040 TI - [Pulmonary toxicity caused by drugs]. PMID- 10687041 TI - [Acute respiratory insufficiency as initial manifestation of brain stem lesions]. AB - We describe three patients with different brainstem lesions (bulbar angioma, bulbar infarct, and Arnold-Chiari malformation) who debuted with acute respiratory insufficiency. Other neurological manifestations had gone unobserved in all three cases. Respiratory insufficiency worsened notably during sleep to the point that mechanical ventilation was required or death occurred (Ondine s curse). The patient with a bulbar angioma is stable with only assisted ventilation by a nasal route at night, with good quality of life. Our conclusions are that: a) central nervous system anomalies need to be investigated as possible causes of respiratory insufficiency when lungs are normal; b) the respiratory control of patients with brainstem lesions should be studied, particularly at night (polysomnography), and c) even when awake-state ventilation is adequate, nighttime assisted ventilation may be required. PMID- 10687042 TI - [Benign metastasizing pulmonary leiomyomatosis. A report of 3 cases]. AB - The benign metastasizing leiomyoma is an uncommon variety of leiomyoma, tumor derived from smooth muscular tissue. The benign metastasizing leiomyoma affects a middle age women, with antecedents of uterine leiomyoma, the pulmonary lesions appeared as a multiple nodules, without systemic affectation. We present three cases of benign metastasizing leiomyoma that de diagnosis was made for biopsy by thoracotomy; and in one case the markers from estrogens' receivers were positive. PMID- 10687043 TI - [20210G/A mutation of prothrombin gene in a patient with deep venous thrombosis ad pulmonary embolism without other risk factors of thrombosis]. AB - A new genetic anomaly predisposing to venous thrombosis was described in 1996, namely the transition of guanine (G) to adenine (A) at position 20210 in the 3 untranslated region of the prothrombin gene. This mutation is associated with high levels of plasma prothrombin and increased risk of thrombotic events in the venous system. We report the case of a man who, lacking known risk factors for thrombosis, suffered a massive pulmonary embolism and deep venous thrombosis in both lower legs. Thrombophilic analysis confirmed that the patient and close relatives were carriers of the heterozygotic 20210G/A variant of the prothrombin gene. Two relatives with the genetic defect had also suffered some type of deep venous thrombosis. PMID- 10687044 TI - [Chylothorax caused by fracture of the humerus and resolution after conservative treatment]. PMID- 10687045 TI - [Peripheral neuroectodermal tumor of the posterior mediastinum in an aged patient]. PMID- 10687046 TI - [Neurogenic pulmonary edema after epileptic crisis]. PMID- 10687047 TI - [Bronchial spasm and eosinophilia related to administration of fosinopril]. PMID- 10687048 TI - [On the question of arbitrary "selection" of genetic code]. PMID- 10687049 TI - [Irreversible changes in the ultrastructure of kinetochores in mitosis, caused by human cytomegalovirus]. PMID- 10687050 TI - [Cellular-population mechanisms of regulation during tumor growth]. PMID- 10687051 TI - [Site-specific cleavage of yeast tRNA(Phe) by derivatives of oligonucleotides bearing bisimidazole groups]. PMID- 10687052 TI - [Crezacin--a new biostimulator of microbiological synthesis]. PMID- 10687053 TI - [Secretion of venom from the common viper (Vipera berus) under in vitro conditions]. PMID- 10687054 TI - [Comparative study of changes in the quantitative characteristics of the frog Rana temporaria cerebellar molecular layer structure when exposed to L-glutamate and NO-generating compounds]. PMID- 10687055 TI - [Analysis of discrete variability of rodents using equilibrium population properties]. PMID- 10687056 TI - [Combination of phosphazide and crixivan inhibit replication of HIV-1 strains that are resistant to azidothymidine]. PMID- 10687057 TI - [The effect of annexins on the contractile activity of the smooth muscles in the rat portal vein]. AB - This study examines the effects of the family of calcium-dependent phospholipids binding proteins (annexins) on the contractile properties of rat portal vein. We developed the new method of annexin's purification from pig stomach muscle. It found that in nanomolar concentrations annexins caused dose-dependent effects on phasic and tonic component of rat portal vein contractile activity. The frequency of spontaneous contractions was increased whereas the amplitude was decreased. At the same time we observed the rise of basal tone level of contractions. We suggested that annexins may change the contractile properties of vascular smooth muscle. PMID- 10687058 TI - Intermittent hypoxia alters hypoxic ventilatory responses. AB - Intermittent hypoxic training (IHT) shows promise for prevention and treatment of some diseases and efficiently produces great advancement in athletic training. We studied (1) hypoxic ventilatory responses (HVR) in supine and sitting positions during normobaric, isocapnic, progressive hypoxia (rebreathing technique) and (2) lung ventilation and gas exchange while breathing ambient air at rest and during 5 min of breathing 11% O2. Duel measurements were made pre- and post-15-day IHT regimen on 12 (experimental) healthy males (24.6 y.o. +/- 1.9 y.o.) and on 6 (control) healthy males (24.2 y.o. +/- 2.3 y.o.) given pseudo-IHT (p-IHT) without decreasing PiO2. IHT involved rebreathing eucapnic (chemically absorbed) air as P(ET)O2 decreased to 35 mmHg, three 6-7 min sessions, three times a day, with 10 min breaks between each session over a 15 day training period. Without IHT, HVRs were the same in sitting and supine positions at low levels of hypoxic challenge (slope one--S1: P(ET)O2 from 110-60 mm Hg) and significantly higher (by 45%) during severe hypoxia (slope two--S2: P(ET)O2 from 60-35 mm Hg). IHT caused an increase in HVR in both sitting and supine positions: S1 by 70 and 100%, S2 by 158 and 200%, maximal lung ventilation by 35 and 78%, respectively. There were no significant changes in the p-IHT group. IHT also caused enhanced respiratory reactions during sustained hypoxia (lung and alveolar ventilation increased by 36 and 22%, respectively). A striking hypoxic ventilatory sensitivity was noted in subjects with hyper-reactive breathing patterns. PMID- 10687059 TI - [The status of the stress-limiting and stress-realizing links in adaptation in newborn rabbits with experimental chronic fetal hypoxia]. AB - The model of chronic intrauterine hypoxia was made according to the method, devised in physiology and experimental medicine laboratory of Institute of Pediatrics, Obstetrics and Gynecology Ukrainian AM Sci. This method consist in dose narrowing of the basal venous columns from cornua of uterus in rabbits on 14 15 days of pregnancy. The obtained experimental results confirm previous clinical date about disturbances in antistress mechanisms of protection under chronic fetus hypoxia. Evidently, this disturbances are the important pathogenetic links in behinding of newborn rabbits in physical development and, especially, in its lethality. PMID- 10687060 TI - [The characteristics of the background neuronal activity of the rostral hypothalamus and an analysis of its alteration under the influence of the stimulation of phylogenetically heterogeneous sections of the cerebral cortex]. AB - In acute experiments on cats under mixed narcosis (ketamine + N2O) we have investigated background firing activity of the rostral hypothalamic neurons and analyzed it modifications due to serial stimulation (100 imp/s, during 5 s) of the prefrontal, cingulate, pyriform cortices and hippocampus. Analysis of the ongoing mean frequency histograms allowed to single out three types of background firing activity with different rhythmical properties. Three types of interspike intervals distribution i.e. asymmetrical (A), symmetrical (S) and polymodal (P) have been distinguished as well. Cortical stimulation was found to transform types of interspike intervals distribution in 15% of hypothalamic neurons. S-type changed into P-type and A-type--into P-type. Author discussed correlation between the character of background firing activity, types of interspike intervals distribution and functional properties of hypothalamic neurons. PMID- 10687061 TI - [The function of the oxytocin-synthesizing system of the hypothalamus in rats with diabetes mellitus undergoing hypoxic training]. AB - The state of hypothalamic oxytocin-synthesizing system in Wistar rats were investigating. The morphometric measurements and immunocytochemical detection of oxytocin-containing cells was used for determining of the functional state of supraoptic nucleus, anterior and posterior-medialis magnocellular subdivisions of paraventricular nucleus. It was established intermittent hypoxic training exert positive influence on rats with experimental diabetes mellitus. This effects depending on increasing synthesis and secretion of hypothalamic oxytocin. Intermittent hypoxic training elevate contents of immunoreactive oxytocin without changing morphometric characteristics in neurons of supraoptic and paraventricular nuclei and median eminence of hypothalamus. In comparison oxytocin contents in these neurons elevade less significance in diabetic rats, but it was observed increasing of nucleolus volume in hypothalamic oxytocin synthesizing neurons. Intermittent hypoxic training of diabetic rats stimulate more significance elevating oxytocin contents in hypothalamic neurons and median eminence that evidence high level activity of hypothalamic oxytocin-synthesizing system. PMID- 10687062 TI - [The circulatory changes as dependent on the type of autonomic homeostasis in divers in dives to a depth of 65 m]. AB - Investigations were performed during 24 simulated air dives under pressure 0.74 MPa in which 16 divers took part. Before, during and after dives intrathoracic pressure, mean arterial pressure, ECG, rheoplethysmogram and intervalocardiogram were recorded continuously with the Valsalva maneuvers. Variables of heart pumping function, peripheral vascular system, phasic heart cycle structure were calculated. For evaluation of autonomic homeostasis variational and spectral analysis of cardiac rhythm were used. In hyperbaria a part of hemodynamic response peculiarities to intrathoracic pressure increase were common to all divers and caused by hyperoxia and increased density of breathing medium, which lead to the appearance of bradycardia, decrease of cardiac output, peripheral vasoconstriction enhancement. Other observed features were determined by alterations in sympathetic and parasympathetic nervous system tonus. These changes were rather complex and highly individual. Our data may benefit evaluation of hyperbaric factor stressful action on blood flow and autonomic nervous system. PMID- 10687063 TI - [The characteristics of the morphofunctional status of 15- to 17-year-old youths based on a comprehensive assessment of the rates of their biological development]. AB - There were studied individual typological peculiarities of the genesis of the genesis of 15-17 years old boys, the state of their respiration and their heart activities. According to these experiments there was stated the possibility of the complex appreciation of the biological tempos of genesis of the senior grades' pupils on the bases of physical and somatic genesis and sexual maturity. As a result of such differentiation of 15-17 year old boys there will appear 3 typological groups--fast, normal and slow types of genesis which quite differ by the majority of grades. Besides these 3 groups differ by the functioning of cardiorespiratory system. Boys with fast tempos of genesis are closer by the functioning peculiarities of cardiorespiratory system to the normal tempos of genesis which are understudy. The group with the slow genesis is quite different from the groups with the fast and normal genesis. PMID- 10687064 TI - [The in-vitro resumption of meiotic maturation and of the first polar body by mouse oocytes at different stages of the estrous cycle]. AB - Resumption of the meiotic maturation and extruded the first polar body of the murine oocytes was registered at 2 and 14 hours under culturing in the medium containing 1.7 mM Ca2+. There were differences between the groups in the percentage of GV oocytes with zona pellucida that acquired competence to undergo germinal vesicle breakdown (GVBD) at 2 hr time point 22.2% of "small" and 32.4% of "large" follicles of mouse at dioestrus and, respectively, 28.1% and 41.8% follicles of mouse at oestrous underwent GVBD. At 14 time point 21.4% of "small" and 30.3% of "large" follicles of mice at mice at dioestrus and, respectively, 26.1% and 43.6% follicles of mouse at oestrous underwent GVBD and (then) extruded the first polar body (FPB). It may be suggested, that acquisition of competence of the oocytes to resume meiosis in vitro depends on the size of follicles and the stage of murine oestrous cycle. Thus, the largest percentage of GV oocytes resuming meiosis was from "largest" follicles of mouse at oestrous. PMID- 10687065 TI - [The balance of elastase and its inhibitors in the vascular tissues of rabbits of different ages in the early stages of experimental Monckeberg-type arteriosclerosis]. AB - The role of the elastolytic system in pathogenesis of vascular diseases was investigated on adult and one-month old rabbits in experimental ergocalciferol induced media calcinosis depending on age aspect. The obtained results indicate that elastase activity was increased in aortic homogenates of one-month old rabbits but not in adult animals. The level of alpha 1-proteinase inhibitor is reduced in one-month old rabbits, and decrease of the alpha 2-macroglobulin content in arterial walls occurs in adults. A higher level of antielastase proteins in both groups of animals in venous vessels is determined. After effect of ergocalciferol in the veins of one-month-old rabbits, differs from the adults, a significant increase of the inhibitor content is observed. The presented results confirm the importance of balance between elastase and it inhibitors in pathogenesis of arteriosclerosis. PMID- 10687066 TI - [Compensatory changes in the acid-base balance under the influence of excess ammonium chloride, hypodynamia and stress]. AB - Two variants of alternative integrative compensatory mechanisms of metabolic reactions in organism developing at the compensated shifts of acid-base balance under the influence of different risk factors have been discovered by the authors. Compensated metabolic acidosis appears with the excess of salt ammonia in the animals ration, at the immobilization stress, hypodynamia, deafferentation and surgical stress. The start reaction of acidosis is lipolysis activation and nonacidified products accumulation at the reactions of tricarboxylic acid cycle are (TAC) inhibited. The compensatory mechanism directed to bind hydrogen ions and prevent pH shifts is activity of glyconeogenesis reactions, transamination, increase the contents ammonia, as well as by increases of restored combinations. Carbohydrates excess in the ration, emotional stress (anxiety expectation) lead to the metabolic alkalosis development at which low hydrogen ion formation and their rapid use in the activated process of peroxide lipids oxidation is compensated with the increase of organic acids formation in glycolysis and TAC as well as by the increase of oxidation properties in tissues. Besides adaptive physiological meaning, the described mechanisms may be the reason for a number of pathological state appearance. The discovery in population and prevention of endogenic risk factors prevention is principle new basis of modern integrated system of human and animals common diseases prevention. PMID- 10687067 TI - [The biological significance of the genetically determined Se-se human blood group and its effect on the antibody formation process in donors immunized with staphylococcal anatoxin]. AB - 82 blood donors have been observed, 63 of them were immunized. Blood group ABO(H), secreting group Se--se and Staphylococcus antibody contents (anti-alpha staphylolysins) were determined in all the donors. It was found out that the donors-secretors with A(II) blood group exhibited the antibody-production increasing. It is supposed that the secreting of group-specific substance A, that has structural elements similar those of staphylococcus into saliva promotes antibody production increase against staphylococcus. The mechanism of such specific stimulation remains to be unknown and requires further studying. PMID- 10687068 TI - [The reaction of regenerating connective tissue to acetylcholine in the dynamics of the denervation-reinnervation process]. AB - The granulation tissue of wounds on upper surface of feet on the 7th day under repeated acetylcholine (Ach) application (0.02 g/1) was studied. In the control group, Ach-induced intensification of inflammatory process and bad condition of regenerating tissue of skin. This pathological reaction increased (fastly appeared ulceration) at once after crushing of right sciatic nerve in the denervated wound. It was less pathological in the contralateral (innervated) wounds. The effect of ACh was positive on later term of neurodystrophic process in the denervated wound and in the contralateral one. Cholinoceptive of granulation tissue of bilateral wound became normal after reinnervation. PMID- 10687069 TI - [The effect of enterosorption on the local immune processes in a toxic lesion of the large intestine]. AB - The local immune homeostasis changed of the colon play a significant role in the pathogenesis of toxic colitis, induced by lead acetate. Chronic toxic colitis is accompanied by a considerable redistribution of plasmocytes-producers of the main immunoglobulins classes in the colon walls towards a primary IgA producers increase, a correlation between them and the local immune reactions tension and destabilization as well. Enterosorbent SCNP-2 influence on a pathological process in the colon under the chronic lead intoxication results in the local immune reactions improvement in the studied organ, its functional and structural elements impairment rate reduction as well as their adaptive properties increase. PMID- 10687070 TI - [Changes in the respiratory functional status of children exposed to the chronic effect of ionizing radiation as a consequence of the Chernobyl catastrophe]. AB - Investigating the functions of the respiratory system some disorders of the ventilation capacity and the lung respiratory functions, external respiration, hypoxic changes in circulatory type at the functional loadings were revealed in children exposed to chronic radiation effects during the postaccident period. It was supposed that an increase in the intensity of free radical processes in the body and the presence of dysadaptation conditions are the basis of changes revealed. PMID- 10687071 TI - [The features of the psychophysiological functions in older preschoolers and younger school-age children and the influence on them of social isolation]. AB - Psychophysiological properties and psychophysiological rating as an integral index of human mental efficiency were measured for elder preschool children, 1-2 nd grade students of primary school and also for elder preschool children and 2 nd grade students of boarding school for orphaned children. It was shown development of psychophysiological properties with age. An influence of social isolation caused to slowing down in development of psychophysiological properties, especially of indices of functional level of system and precision of reaction on moving object. PMID- 10687072 TI - [The age-related characteristics of human mental work capacity]. AB - The aim of investigation was to study the ageing peculiarities of human mental working capacity with mathematical models. Were examined 150 women and 125 men in five aged groups: 13-18, 19-29, 30-39, 40-49, 50-60. The human mental activity was presented with special computer's tests. The regression ageing models for assessment of mental working capacity is proposed. The study showed that high level of human mental working capacity is determined with the decreasing of mental experience variation and the increasing of speed of remaking the information. PMID- 10687073 TI - [The humoral reactions of bioincompatibility in the early period after the heterotopic autograft of veins into the coronary bed and the implantation of synthetic heart valves]. AB - Humoral manifestations of bioincompatibility were studied at early stages (during several hours and days) after operations using non-peculiar to heart trans- and implants: aortocoronary bypass at 102 patients with coronarosclerosis and implantation of artificial cardiac valves at 145 patients with endocarditis. In the various postoperation stages the formation of the structurally and functionally different types of the nonclonal specific autoprecipitins to autological membranocellular components was revealed at recipients by methods of double immunodiffusion and immunoelectrophoresis. These autoprecipitins such as: early, activated and tardly, synthesized under effect of alien agent--may be useful as diagnostic and prognostic indicators of bioincompatibility and its clinical consequences at the earliest period after cardiac operations using trans and implants. PMID- 10687074 TI - [The effect of a grape extract on the contractile activity of the myocardium and on the coronary flow of the isolated guinea pig heart]. AB - Effects of one of the fraction of grape extract on myocardial contractility and coronary flow of isolated guinea pig heart have been investigated. It was shown that the administration of grape extract led to myocardial contractility index is increase by 77%, coronary flow increase by 13% and oxygen consumption increase by 30%. Inhibition of nitric oxide synthesis by L-NMMA resulted in decrease of grape extract-induced coronary flow increase. These data support of isolated strips investigation that grape has a compounds which induced endothelium-dependent vasodilation of coronary bed. They indicate that these substances have the strong cardiostimulating effect. PMID- 10687075 TI - [The length-force dependence of vascular smooth muscles and the nitric oxide system in a chronic deficiency of mesostriatal dopamine]. AB - This study investigated the influence of the chronic cerebral dopamine deficiency after 6-hydroxydopamine lesions of dopaminergic mesostriatal system and NO pathways on the length-tension relation in vascular smooth muscles to their distension. Experiments were performed on isolated strips of rat portal vein. The results indicate that the attenuated contraction responses and the increased stiffness of vascular smooth muscles to the distension in a chronic mesostriatal dopamine deficiency. It was found that these responses may be, in part, normalized after a long-term L-arginine administration. The same changes were marked in the responses of intact vascular strips after the inhibition of NO synthase activity and L-arginine perfusion. It has been suggested that, in a chronic mesostriatal dopamine deficiency, the reduced vascular reactivity was due to the decreasing of NO synthesis by endothelium and L-arginine could be used in the treatment of these vascular disorders. PMID- 10687076 TI - [The nonoliguric form of kidney failure in the posthemorrhagic period]. AB - At research of rules of deviations of the parameters homeostasis for patients with bleedings we pursued to find dependence of the parameters of homeostasis from function of kidneys. The researches were carried out in the early posthemorrhagic period on an example of inspection of the patients who were operated concerning gastrointestinal and intraabdominal hemorrhages. Is revealed, that the renal insufficiency, which arose at 23.21% surveyed, proceeded without oliguria--in the neoliguric form. It was characterized by normal speed diuresis, despite of the essential damages of the function of nephron: infringement of a primary filtration in glomerulus, reabsorption of water and electrolytes in tubules, restriction of excretion H+. Normal diuresis in such cases was explained by the early and duly beginning of the infusion therapy, owing to what, probably, the infringement of passableness tubules was prevented. Despite of it, the patients with the damaged function of kidneys require the careful control of spent therapy because of an opportunity of dangerous deviations of the parameters homeostasis of norm. Confirmation it became the reduction, fixed at such patients, clearance of electrolytes, infringement of the extraction of acid by kidneys. PMID- 10687077 TI - [The adaptation of youth with different rates of morphofunctional development to physical loading]. AB - There were studied morphofunctional peculiarities of biological development of 15 17-year-old boys' organisms on the bases of complex use, for this, of the level of physical development, rates of sex maturity and somatotype. On that base there were studied the individual peculiarities of their adaptation to the physical loads, which was directed to the development of endurance. It was ascertained, that the boys of the examined age groups, based on the complex differentiation are divided into 3 basic groups--fast, normal and slow rates of genesis. These typological groups differ by the measure and quantity of periods of long-term adaptation, and by the nature of adaptable changes in the organisms which are studied as a result of influence of long-lasted dosing loads, directed to the development of endurance. PMID- 10687078 TI - [The effect of the duration of coronary occlusion on the healing of an experimental myocardial infarct]. AB - Increasing the period of myocardial ischaemia prior to conduction of reperfusion adequality induces complicated healing of experimental myocardial infarction, thereby resulting in postinfarctional aneurysm of the heart. Complicated healing of myocardial infarction upon of coronary blood re-circulation after 3 or more hours of ischaemia, happens due to disbalance of necrotic and reparative processes in the infarct zone. Conduction of reperfusion induces activation of necrotic processes with retention and slowdown of reparative processes. PMID- 10687079 TI - [The comparative effect of magnetic and laser irradiation of the liver and blood on the bile-secretory function in rats]. AB - The influence of percutaneous magnetolazer irradiation of the blood and liver on the bile secretion, general bilirubin excretion and its fractions, cholesterol and bile acids rate was studied experimentally on male rats. A laser generator "Luch-2" with a magnetic nozzle (wave-length 0.82 mm, power density 0.178 W.cm-2, magnetic field tension 30-35 mT, the course s 2 daily procedures) was used. Bile secretion intensity was noted to depend on the dose. Maximal bile secretion and its basic components were observed during the liver irrigation with an exposition for 120 s and blood ones with an exposition for 240 s respectively. Bile secretion rate was higher during magnetolazer influence on the blood. Liver irrigation was associated with an increased secretion of the conjugated bilirubin. Thus, there are various stimulant mechanisms of the liver function activity in the course of magnetolazer influence on the liver and blood. PMID- 10687080 TI - [The protective activity of anti-Proteus blood preparations]. AB - Human blood anti-Proteus preparation activity has been studied on the model of Proteus etiology sepsis in white not pedigree mice. It has been found out that anti-Proteus plasma and anti-Proteus immunoglobulin with antibody titre 1:80 possess marked therapeutic properties. The obtained results of the experimental studies are the grounds for clinical studies carrying out with the use of blood anti-Proteus preparations in combined therapy of the patients with diseases of Proteus etiology. PMID- 10687081 TI - [The effect of aromatic plant substances on the status of oxidative-reductive enzymes in a chronic experiment]. AB - In chronic experiment (during 3 months) was studied the influence of various vegetable aromatic substances (VAS) contents in the air upon oxidative restoration enzymes activity in experimental animals (Wistar line male rats). On the base these experiments' results may be made a conclusion that the lack of VAS in the air involves changes in the most important enzymes of glycolysis and pentosophosphatic cycle. Provision of the atmosphere with essential lavender oil in concentration 0.58 mg/m3 (natural concentration) may correct such disturbances. PMID- 10687082 TI - [The effect of divalent cations on pepsinogen extrusion by permeable isolated gastric glands]. AB - The model of permeable gastric glands obtained by its incubation with digiton (15 mg/ml) had investigated. It was shown, that stimulated influence of Ca2+ ions on pepsinogen extrusion completely or partly reproduced by cations of transient metals. Stimulated effects of cations of metals (5 x 10(-4) M) decrease in such order: Ca2+ > Cd2+ > Mn2+ > Co2+ > La3+. Nevertheless, Co2+ and La3+ in the same concentrations inhibit Ca2+ stimulated extrusion of pepsinogen by permeable isolated glands. Stimulated effect of Cd2+ and La3+ on pepsinogen extrusion in native glands was not reproducible. Besides these cations inhibit carbacholine stimulated extrusion in native glands. It's concluded, that influence of metal cations on secretory function of gastric glands to a great extent is determined by their property to penetrate into secretory cells and cooperate with their plasma membranes. PMID- 10687083 TI - [The effect of sodium alpha-glutarate on enzymatic transamination activity and on succinate dehydrogenase in rats with different resistances to hypoxia]. AB - It have been found that intraperitoneal alpha-ketoglutarate injection (20 mg/100 g body weight) results in increase in the influence of cholinergic regulation mechanisms. It also results in increase of aminotransferase activity on background of the decrease of succinate dehydrogenase activity in liver and pancreas tissues and in small intestines mucous. Activity of transamination enzymes and succinate dehydrogenase activity is much higher in the case of rats with high hypoxia resistance, alpha-ketoglutarate injection results in increase of transamination enzymes activity in the organisms of rats with low resistance to hypoxia up to the control level of rats with high resistance, and simultaneously increases rats resistance to hypoxia. Effect of alpha ketoglutarate injection on the energetical exchange in the tissues with different parasympathetic dependence taking from the animals with different hypoxia resistance is suppressed by blockade of M- and H-cholinoceptors. PMID- 10687084 TI - [The physiological and biochemical indices of the disposition of rats to alcohol use]. AB - Activity of aldehide dehydrogenase (ALDH), alcohol dehydrogenase (ADH) in cytoplasmic fractions of brain structures (hypothalamus, midbrain, new cortex) and in blood serum as well as the content of noradreneline (NA) and dofamin (DA) in the mentioned structures and blood of rats preferring ethanol (PE) and rejecting ethanol (RE) has been investigated. ALDH isoforms have been revealed in rats preferring ethanol (PE) and these rejecting it (RE). The activity of the revealed forms of ALDH is higher in PE rats that in RE rats. PE and RE rats do not differ from one another as to ADH activity. It is shown that the NA content is decreased in PE rats and the DA level is increased in a number of brain structures and in blood as compared to RE rats. PMID- 10687085 TI - [The effect of prolonged physical loads on the cardiovascular system of middle school-aged pupils]. AB - The cardiovascular system of schoolchildren at the age of 11-15 was studied as affected by systemic of long lasting physical loads of schoolchildren. In the trained children contrary to adults the respiratory arrhythmia is observed, that evidences for nonadequacy of the physical load to the functional state of myocardium. In the cardiovascular system of the trained schoolchildren under the effect of constant training there occurs a series of functional changes which increase adaptation to physical loads, that is manifested in a decrease of the systolic and diastolic pressure, the lowering of the systolic index and heart rate, and in increase in mechanical and electromechanical systoles, the amplitude of the teeth R and T. The detected changes in the cardiovascular system are due to the value of trained loads and their nonadequacy to the functional state of myocardium. PMID- 10687086 TI - [The effect of a cryoextract of the chorion on the cellular reaction of an inflammatory focus]. AB - On the model of carrageenan-induced acute aseptic peritonitis in rats it is shown that in inflammation on the background of administration of chorion's cryoextract a marked inhibition of neutrophilic reaction and stimulation of macrophagic fibroblastic one, and earlier ending of inflammation is observed, i.e. cryoextract of chorion has an expressive anti-inflammatory action which is mainly realized by inhibition of pathologic reactions and stimulation of protective ones in the blood. PMID- 10687087 TI - [Structural and functional changes to the lungs and the lipid metabolic status in cardiogenic shock]. AB - The functional and structural manifestations of lipid metabolism disorders in the lungs were studied in comparison with the changes of fatty acid composition of lipids of pulmonary tissue, venous and arterial blood, expirated air condensate and sweat during the development of cardiogenic shock in patients with myocardial infarction. It has been established that the lungs accumulate neutral lipids and lose phospholipids and polyunsaturated fatty acids. These changes are combined with intracellular lesion of osmiophilic lamellated corpuscles and disturbance of their excretion into alveolar cavities. At the same time the increase of polyunsaturated fatty acids content in expirated air condensate and sweat of patients is noted can be related to the elimination of the excess of these substrates from intercellular space. PMID- 10687088 TI - [The effect of internal and external irradiation on thyroid gland function in experimental animals]. AB - We are investigated influence of different kinds of radiation (internal--131I, external--gamma and roentgen irradiation) in equivalent doses (5 Gy) on the thyroid function of experimental animals. Revealed distinctions in the dynamic development of the function changes after action different types of radiation. PMID- 10687089 TI - [Free-radical processes under different conditions of body oxygen allowance]. AB - Free radical processes (FRP) in mammalian organism are oxygen-depended. The review is devoted to the analysis of pro- and antioxidant processes in mammalian tissues under different oxygen supply conditions. There are described: sources of free radicals; hyperproduction of FRP under hyperoxia and hyperbaria; the role of free radicals in the adaptation to chronic hypoxia (high altitudes, barochamber, chronic heart and lung diseases); production of active oxygen species during the reoxygenation of preliminarily hypoxic tissues; FRP under intermittent hypoxic training; role of FRP in the chemoreception of oxygen. The special attention is paid to three main factors underlined the FR production in hypoxic conditions: the speed of hypoxia increase, the degree of hypoxia and the time of hypoxic exposure. PMID- 10687090 TI - [Gene therapy: ethical aspects and problems of genetic safety]. AB - Gene therapy (GT) is based on the introduction of various genetic constructions in the human organism. Rapid development of GT technologies and the increasing tendency to apply them in therapy for certain human diseases has created a number of ethical problems. These first and foremost concern direct intervention in the genome of germline cells, which may change the genome of further generations. According to current views, such manipulations are ethically unacceptable. Introduction of genetic constructs in somatic cells with therapeutic purposes (somatic GT) is considered permittable in principle but requires certain procedures to ensure the genetic safety (causing no damage) of both the patient and other people. The latter hazard is associated with a possibility of uncontrolled spreading of viral (especially retroviral) constructions used in many GT protocols. These and other ethical and genetic safety problems of modern GT are discussed. PMID- 10687091 TI - [Analysis of differences in the structure of ribosomal DNA from two sibling species: Drosophila melanogaster and Drosophila simulans]. AB - Southern-blot hybridization studies have revealed structural differences within the ribosomal DNA clusters of two sibling species, Drosophila melanogaster and Drosophila simulans. The approach used was shown to be suitable for taxonomic identification of the species examined. PMID- 10687092 TI - [Conjugative transfer of a plasmid from Escherichia coli to various strains of the order Actinomycetales]. AB - The conjugal transfer of autonomous and integrative plasmids from the donor strain Escherichia coli S17-1 to strains of genera Actinomadura, Arthrobacter, Kitasatoa, Micromonospora, Nocardia, Rhodococcus, Saccharopolyspora, and to 16 strains of the genus Streptomyces was demonstrated. The status of plasmids in recipient strains and the stability of their inheritance were analyzed. Plasmids constructed for strains of the genus Streptomyces were shown to function in a large number of strains belonging to the order Actinomycetales. The well developed system of Streptomyces vector molecules and cloned genes of antibiotic biosynthesis allows their transfer to those microorganisms for which conventional techniques of plasmid transfer by regenerated protoplast transformation or electroporation have not been developed or are inefficient. PMID- 10687093 TI - [Double crossing over: elementary events and the sequence of their occurrence]. AB - Analysis of the crossing over increment in the structurally normal chromosome of Drosophila caused by a rearrangement in nonhomologous chromosome (interchromosomal effect on crossing over, IEC) was carried out based on the author's personal and literature data. The IEC in the left arm of chromosome 2 caused by inversions in chromosomes X and 3, as well as the IEC in X chromosome caused by inversions in chromosomes 2 and 3, were examined. The IEC-induced increment of crossing over results from the increase of the number of double exchanges under the constant or reduced number of single exchanges. Tetrad analysis showed that the given alternation of the crossing over processes could occur only in the case of conversion of the tetrads with single exchanges into the tetrads with double exchanges. In other words, the events leading to the formation of double exchanges occur consecutively. The borders of the IEC-induced double exchanges can be seen all over the chromosome body. However, the IEC induced increase of chromosome recombination length occurs only in the proximal region (in rare cases, in proximal and distal regions) of the chromosome arm. This means that a double exchange is formed when the first event with predominant location in the middle of the arm is supplemented with the second event predominantly localized at the arm T end, most frequently in the proximal region. The pattern of the IEC-induced double exchange formation can be satisfactorily described in terms of the contact model of the crossing over. According to the model, an elementary crossing-over event is the local contact between the homologues. Neither single exchange nor a double-stranded DNA break can serve as an elementary event in the process of any multiple exchange formation. PMID- 10687094 TI - [Effect of inactivating various components of the signal pathways of the tumor suppressor p53 on genomic stability]. AB - To evaluate the role of different p53-regulated signaling pathways in the control of genomic integrity, we studied the frequency of changes in chromosome number and structure of cells of the sublines of mouse primary embryonic fibroblasts with the "knocked-out" genes for proteins p53, p21WAF, pRb, and p19ARF. Protein p21WAF is transactivated by p53 and is responsible for the cell block in the G1 phase of the damaged cells; protein pRb is a target for p21WAF which controls the G1-S-phase transition; and p19ARF protein is responsible for p53 activation in cells with certain anomalies. Inactivation of either of the studied genes proved to increase significantly the frequency of changes in the karyotype. However, the resultant chromosome instability differed: the frequency of the chromosome breaks, both spontaneous and induced with ethylmethane sulfonate (EMS), was in cells with inactivated p53 and lowest in cells with inactivated pRb. These distinctions were not caused by a different effect of various gene inactivation on the cell cycle progression: in all sublines, the cell block in G1 was abolished and the checkpoint function in G2 remained normal. However, the induction of apoptosis in EMS-treated cells differed in the studied sublines. The lowest number of apoptotic nuclei were determined in p53-/- cultures, whereas the highest were in the Rb-/- cultures. It is apparent that the degree of genetic instability is determined by a combined effect of apoptosis and abnormal regulation of the cell-cycle checkpoints. PMID- 10687095 TI - [Association of mutations in cytochrome b and NADH-dehydrogenase 5/6 mitochondrial DNA genes from the sable (Martes zibellina L.)]. AB - RFLP analysis of amplified genes for cytochrome b and NADP dehydrogenase 5/6 of sable mtDNA was carried out. Polymorphism was recorded in the first and the second fragment using six and four enzymes, respectively. This allowed us to identify seven mtDNA haplotypes forming two clusters, A1-A3 and B1-C2. The divergence between the clusters was 0.0112-0.0164. An association between mutations of genes for cytochrome b and NADP dehydrogenase 5/6 was established. This association can be explained by high mutability of the fragments containing the associated mutations. PMID- 10687096 TI - [Intrapopulation autosomal polymorphism in the common vole Microtus arvalis from the Transcaucasian region]. AB - The broad autosomal polymorphism in form obscurus of common voles Microtus arvalis from the Transcaucasian region that is associated with the variation of subtelocentric chromosome pair 5, as well as the mechanism and evolutionary significance of this polymorphism, are discussed. Based on the morphological analysis of heterozygotes for chromosome pair 5 after differential G-, C-, and Ag NOR-banding and on the measurements of homologues, the following conclusion has been made. The occurrence of the acrocentric chromosome 5 is the result of a double chromosomal rearrangement: a pericentric inversion and a duplication of the chromosomal material. The mutation has been found throughout the entire territory of Armenia. In spite of such a wide distribution, the mutation frequency in populations is extremely low. Neither a definite pattern of geographic distribution nor a clinal variation was found for this mutation. This mutation is likely to occur independently in different M. arvalis populations and is apparently neutral. Homozygotes for chromosome pair 5 are described for the first time. PMID- 10687097 TI - [Genetic-demographic processes in urban populations of the Ukraine in the 90's. The marital structure of the Poltava population]. AB - Analysis of records of the marriages that were contracted in the city of Poltava revealed an increase in outbreeding between 1960 and 1985 in the Poltava population. This was expressed in increased ethnic diversity, proportion of interethnic marriages, migration range, and the parent-offspring and average marital distances, whereas the proportion of the indigenous ethnic group (Ukrainians) decreased. By 1995, outbreeding decreased: the ethnic composition of the population became more homogeneous (98% Slavic), the proportion of Ukrainians and the frequency of monoethnic marriages increased, and the migration range decreased. During the period studied, the population became more panmictic with respect to ethnicity and birthplace, with the social and professional subdivisions of the population remaining unchanged. PMID- 10687098 TI - [Structure and diversity of the mitochondrial gene pool of the aboriginal population of Tuva and Buriatia from restriction polymorphism data]. AB - The populations of Tuvinians (N = 36) and Buryats (N = 105) were characterized by using the data on mitochondrial DNA (mtDNA) polymorphism. The gene pools of both ethnic groups possessed the mtDNA types belonging to the four main haplogroups, A, B, C, and D, found only in the indigenous populations of Asia and America. The total frequencies of the A, B, C, and D haplogroups in Tuvinians and Buryats were 72.3% and 52.4%, respectively. These values, along with the frequency for Altai populations (57.2%), were highest in the Asian populations studied, indicating that the populations Southern and Eastern Siberia can be considered as ancestral relatives to the ethnic groups of the New World. Analysis of the mtDNA region V polymorphism showed the presence of 9-bp deletion and 4-bp insertion in both populations with frequencies respectively of 13.9 and 5.56% in Tuvinians and 4.8 and 1.9% in Buryats. The frequency of the +AvaII/8249 variant was 11.1% in Tuvinians and 3.81% in Buryats. Analysis of the association between the region V deletion-insertion polymorphism and certain restriction haplogroups pointed to repeated and independent emergence of the 4-bp insertion in Siberia. PMID- 10687099 TI - [Polymorphism of exon 4 in the CANP-3 gene in patients with primary myopathies]. AB - The structures of the gene for calpain (CANP-3) and of the DMD gene were analyzed in patients with primary myopathies [limb-girdle muscular distrophy (LGMD) and Duchenne-Becker myodystrophy (DBM)] from various regions of Russia. Via amplification of DNA isolated from the peripheral blood lymphocytes of 74 patients, extended deletions were found in 18 out of 55 patients with DBM. In none of the 19 patients with LGMD, were extended deletions in the CANP-3 gene found. In most patients with LGMD, the amplification of the promoter region and exons 1, 2, 3, 4, 5, and 6 of the CANP-3 gene yielded a single product of corresponding length, but in six patients (three sib pairs), amplification of exon 4 of the CANP-3 gene yielded two products of different size. The following single-strand conformation polymorphism (SSCP) analysis revealed a pronounced polymorphism of exon 4 of the CANP-3 gene in 14 out of 19 patients with LGMD. This structure of exon 4 of the CANP-3 gene was found neither in 16 patients with DBM who had deletions in the DMD gene nor in 16 patients with DBM who had no deletions in the DMD gene. PMID- 10687100 TI - [Spontaneous mutants of Alteromonas espejiana, resistant to kanamycin and bleomycin]. AB - An attempt was made to induce insertions in marine bacterium Alteromonas espejiana Bal-31 (Ae) using the TnphoA transposon. The Ae mutants selected on a kanamycin-containing medium after conjugation of the Ae with the transposon donor, Escherichia coli SM10(pRt291), were resistant not only to kanamycin (Kn), but also to bleomycin (Bm), and were sensitive to tetracycline. Although the mutants were phenotypically similar to insertion mutants, the mutations appeared to be spontaneous. The sensitivity of spontaneous Kmr Ae mutants selected at various Km concentrations to Bm was investigated. The mutants selected at low Km concentrations were resistant to Bm, whereas those selected at high Km concentrations were sensitive to Bm. The possible mechanisms underlying the dual resistance to Bm and aminoglycosides in bacteria are discussed. PMID- 10687101 TI - [Prescription of cardiovascular drugs for non-authorized indications]. PMID- 10687102 TI - [Systematic treatment of acute myocardial infarction with primary angioplasty. Early clinical and angiographic results and after 6 months]. AB - Starting in 1995, at our institution all patients with acute myocardial infarction (AMI) who gave informed consent were treated by primary percutaneous transluminal coronary angioplasty (PTCA) without limitations in entry criteria. This report presents early and six-month clinical and angiographic results of the 720 patients (77% male, median age 64 years) treated by direct PTCA between January 1, 1995 and July 31, 1998. On admission, 33% of patients were in Killip class > 1, and 101 patients (14%) were in early cardiogenic shock. Optimal acute angiographic success (TIMI grade 3 flow with residual stenosis < 30%) was achieved in 683 patients (95%). Primary or unplanned stenting of infarct related artery (IRA) for a suboptimal or poor angiographic result after primary PTCA was performed in 454 patients (63%). The mean time from hospital arrival to recanalization was 62 +/- 28 min. At 30 days, the mortality rate was 4.9% (1.8% in Killip class < 4 patients and 24% in patients with cardiogenic shock). The reinfarction rate was 1.2%. At 30 days, coronary angiography showed restenosis or reocclusion of the IRA in 55 patients (8.9%). During the six-month follow-up (30 180 days), there were 11 deaths (1.5%) and 2 non-fatal reinfarctions (0.3%). At six months, the IRA patency rate was 95%, while the mean ejection fraction improvement in 422 patients with paired ventriculograms was 7%. Recurrent ischemia occurred in 144 patients (20%) and resulted in 7 deaths, 11 non-fatal reinfarctions and 126 repeat targeted vessel revascularization. CONCLUSIONS: The major finding of our experience is that direct coronary angioplasty may result in excellent early and late outcome in a population without limitations in entry criteria. The low mortality and the few recurrent myocardial ischemic events are connected with the high patency rate at 6 months. The extensive use of stents improves the angiographic results and the clinical outcome. PMID- 10687103 TI - Heart transplantation in ischemic heart disease when recipients are older than 55 years and donors older than 50. AB - The purpose of this article is to analyze patients affected with ischemic cardiomyopathy, older than 55 years, who have undergone heart transplantation. We conducted a retrospective analysis comparing clinical course and outcome in patients whose donor age was > or = 50 years (Group A) with patients who had younger donor heart (Group B). Group A was composed of 25 patients, 55 to 68 years old (27.4% of the patients), 20 males and 5 females; Group B was composed of 68 patients, 55 to 66 years old, 65 males and 3 females. Mean donor age in Group A was 54.7 years old (range 51-61), while in Group B it was 29.5 years old (range 9-49). Operative mortality was 16% (4 cases) and 12% in Group B (8 cases) p = ns. Total mortality in Group A was 24%, or 6 cases: 2 graft failures, 1 infection, 1 neoplasm, 1 multiorgan failure, 1 ischemic heart disease; in Group B it was 27%, or 18 cases: 2 cerebrovascular accidents, 4 graft failures, 3 infections, 5 neoplasms, 3 multiorgan failures, 1 acute rejection, p = ns. Coronarography was performed in 51 patients, 14 in Group A (10 cases normal, 3 with irregularities, and 1 case with a critical stenosis of the circumflex artery; 37 in Group B (32 cases were normal, 3 had irregularities and 2 had critical stenosis in a coronary artery). In conclusion, we emphasize that extending donor age in recipients older than 55 years of age does not determine a higher risk and mortality. PMID- 10687104 TI - Diagnosis and early surgical management of traumatic thoracic aortic disruption. AB - OBJECTIVE: Traumatic rupture of the thoracic aorta is a major cause of death. Survival greatly depends on early diagnosis, degree of injuries to other districts and timing of repair. To address the controversial aspects of this condition, we retrospectively reviewed our experience. METHODS: Between April 1984 and December 1998, 39 patients (31 males, 79%), with a mean age of 33 +/- 7 (range, 17 to 59 years), underwent surgical repair at our institution. Final diagnosis of aortic disruption was achieved in 33 patients (85%) by aortogram, and in 6 (15%) by transesophageal echocardiography (TEE) alone. Four patients (8%) had a false negative chest X-ray on admission. Twenty-four patients (61.5%) had additional major injuries to other districts (n = 4, cranial trauma; n = 13, cranial trauma + pelvic fracture; n = 5, lesions to abdominal viscera; n = 2, lesions to abdominal viscera + pelvic fracture). Surgical techniques included simple aortic cross-clamping in 7 patients (18%), partial femoral-femoral bypass in 17 (44%), and partial left heart bypass in 15 (38%). Two patients underwent direct aortic suture (5%), whereas 37 (95%) had interposition of a vascular graft. RESULTS: Three patients (8%) died after major hemorrhaging during the early phases of our experience. Paraplegia occurred in 1 patient (2.5%) in the single aortic cross-clamping group. There was no morbidity directly attributable to the administration of heparin for cardiopulmonary bypass. CONCLUSION: Although aortography is still the gold standard to achieve diagnosis, the use of TEE as a method of detecting traumatic injury to the thoracic aorta appears feasible in critical patients, advantageously saving time. With a meticulous surgical approach and the use of an effective method for distal aortic perfusion during repair, it is possible to achieve good outcomes. PMID- 10687105 TI - The limitations of echocardiography in the overall diagnosis of the morphological lesions associated with infective endocarditis: comparison of echocardiographic and surgical findings. AB - Echocardiography is commonly accepted as the method of choice for the non invasive diagnosis of vegetations and other lesions associated with infective endocarditis. To assess the accuracy of echocardiography in the overall diagnosis of the morphological cardiac lesions we retrospectively analyzed and compared the preoperative echocardiographic data with the surgical findings of 120 consecutive cases operated for infective endocarditis. Transthoracic echocardiography (TTE) was used in 60 cases (51 with native and 9 with prosthetic valves), both TTE and transesophageal echocardiography (TEE) in 50 (26 with native and 24 with prosthetic valves) and only TEE in 10 patients who underwent emergency surgery. The echocardiographic diagnosis was correct in all the cases, but incomplete in 26 (16 with native and 10 with prosthetic valves). Most of the incomplete diagnoses occurred regarding vegetations, perforations of the valvular leaflets and perivalvular abscesses. There were no differences between aortic or mitral valves (14/66 vs 11/60; p = ns), native or prosthetic (16/79 vs 10/37; p = ns), TTE or TEE (13/60 vs 13/60; p = ns); however, TEE was performed in more complex cases and in severely ill patients. In six of the incomplete diagnoses, echocardiography preceded surgery by one week or more, and in six the mistakes were not confirmed by the reviewer. In conclusion, our study suggests that an echocardiographic diagnosis of endocarditis may be correct but sometimes incomplete. In patients without prosthetic valves who have a technically-adequate transthoracic echocardiogram, transesophageal echocardiography is not indispensable but should be chosen from time to time. However, the patients with endocarditis and no contraindication to the transesophageal procedure should undergo both transesophageal and transthoracic echocardiography before surgery in order to obtain as much and the most definite information possible. An echocardiographic study should be repeated just before any surgical procedures in patients with active endocarditis. Finally, it needs to be emphasized that the training and clinical judgement of the operator performing the study are important elements determining the results of echocardiographic study. PMID- 10687106 TI - QT dispersion and early arrhythmic risk during acute myocardial infarction. AB - BACKGROUND: QT dispersion (maximal minus minimal QT interval calculated on a standard 12-lead electrocardiogram) has been suggested to reflect regional variations of ventricular repolarization and to provide a substrate for reentry ventricular arrhythmias. In this study we evaluate QT dispersion in patients with acute myocardial infarction and assess its relation with early severe ventricular arrhythmias. METHODS AND RESULTS: We studied 101 patients with acute myocardial infarction and a control group of 97 healthy subjects. We determined QT and QTc dispersion on the electrocardiograms performed 12 hours and 3 and 10 days after the onset of symptoms in myocardial infarction patients and on the control group. The average values of QT and QTc dispersion (measured hereafter in milliseconds, ms) were as follows: 70.5 +/- 42.5-87 +/- 46.6 (after 12 hours), 66.5 +/- 37.8 76.9 +/- 43.5 (on day 3), 68.9 +/- 42-76.3 +/- 43.8 (on day 10) and 44 +/- 13.4 54.2 +/- 16.3 (in control group). We observed statistically significant differences in QT and QTc dispersion between the electrocardiogram of normal subjects and each of the three electrocardiograms performed on patients with infarction (p < 0.0005, p < 0.005). We recorded a greater QT dispersion in patients with anterior infarction with respect to those with inferior/lateral infarction (79 +/- 38.6 vs 65.2 +/- 43.16, p < 0.05) and in patients with ejection fraction < 45% (93.1 +/- 28.4 vs 68.3 +/- 34.1 p < 0.005). During the first three days, QT dispersion did not differ in patients treated with thrombolytic agents with respect to those who were untreated, while on day 10 untreated patients showed higher values (74.9 +/- 45.3 vs 60.5 +/- 37.7, p < 0.05). Creatine kinase peak level, sex and age of the patients did not influence QT dispersion. Thirteen patients (12.8%) developed severe ventricular arrhythmias within 72 hours after infarction: 8 patients (7.9%) had ventricular fibrillation and 5 patients (4.9%) had sustained ventricular tachycardia. We found higher early QT and QTc dispersion values in patients who developed severe ventricular arrhythmias (108.8 +/- 63.2 and 125.8 +/- 68.5) with respect to patients who did not (63.3 +/- 32.9 and 80.8 +/- 38.9, p < 0.0005, p < 0.0005). CONCLUSIONS: Our data suggest that QT dispersion: 1) increases during acute myocardial infarction; 2) peaks in the early hours after symptom onset; 3) drops late after infarction in patients treated with thrombolytic agents; 4) is associated with early severe ventricular arrhythmias. PMID- 10687107 TI - [Improvement of HF: initiative to increase awareness and improve the management of patients with heart failure in Europe]. AB - Heart failure is a major healthcare problem, with possibly over 10 million patients in Europe suffering from the syndrome. Its prevalence is increasing partly because the proportion of elderly population is likewise increasing. The guidelines on the diagnosis and treatment of heart failure have been published by the European Society of Cardiology and represent a landmark for judging current medical practice. Evidence suggests that heart failure is frequently underdiagnosed and undertreated, and the majority of patients may not be receiving appropriate care. Both primary healthcare physicians and hospital cardiologists are responsible for proper diagnosis and appropriate treatment of patients with heart failure. The "Improvement of HF" initiative has been developed by an International Advisory Board to increase awareness and improve the management of heart failure among primary healthcare physicians in Europe. The study first investigates how primary healthcare physicians perceive heart failure should be diagnosed and treated, and examines the availability of required resources. Subsequently, primary healthcare physicians will be interviewed to assess their actual practice by reviewing relevant case notes, covering diagnosis and treatment. The program will consist of a research phase and an educational phase. For the research phase, 10 regional centers (including both urban and rural areas) in each of the 14 participating countries in Europe will be identified, with each center randomly selecting 10 primary healthcare physicians. Each physician will supply case notes of 9 of his/her patients with diagnosed heart failure or at high risk of having heart failure. The results of the survey would identify any need to change not only perceptions but clinical practice in the diagnosis and treatment of heart failure and will be used to organize an educational program. A further study is planned in order to assess the impact of the initiative. PMID- 10687108 TI - [Improvement of prognosis in idiopathic dilated cardiomyopathy: role of early diagnosis and optimized medical treatment. Study Group on Heart Muscle Diseases]. AB - BACKGROUND: The aim of this study was to assess the extent to which a more widespread use of new effective treatments for heart failure associated with earlier detection of the disease may have contributed to enhancing the prognosis of idiopathic dilated cardiomyopathy (IDC) patients over the past 20 years. METHODS: Heart transplant-free survival curves were analyzed in 343 IDC patients, prospectively enrolled from January 1, 1978 to June 30, 1997 in the Heart Muscle Disease Registry of the Cardiology Department in Trieste (94 enrolled between 1978 and 1987, Group 1; 249 between 1988 and 1997, Group 2). At enrollment, 91 patients had no heart failure symptoms (NoHF), whereas the remaining 252 showed HF of recent (HF < or = 6 months, n = 132) or non-recent (HF > 6 months, n = 120) onset. RESULTS: In comparison to Group 1, Group 2 was treated more frequently with ACE-inhibitors and beta-blockers (p < 0.0001) and showed a better long-term survival (p = 0.0034), resulting from a reduction of death for refractory HF or need for heart transplant (p = 0.011). Conversely, the risk of sudden death did not significantly differ between the two groups. NoHF, HF > 6 months and HF < or = 6 months groups were similarly treated with ACE-inhibitors and beta-blockers. Long-term survival was better in patients without HF than in those with overt HF (p = 0.0015). As compared to Group HF > 6 months, Group HF < or = 6 months had a poorer one-year prognosis (p = 0.045), related to the presence of a subgroup of patients with refractory HF and need for heart transplant, but showed a better survival rate over the following years (p = 0.015). Over the two subsequent decades of enrollment, a significant improvement in patient survival was observed within Groups NoHF (p = 0.03) and HF > 6 months (p = 0.01), but not in Group HF < or = 6 months. CONCLUSIONS: Over the past 20 years, the increasing use of ACE inhibitors and beta-blockers in IDC was associated with a significant improvement in long-term survival, resulting from a reduction in mortality for refractory heart failure or need for heart transplant. In addition, early diagnosis may have contributed significantly to enhancing the prognosis of IDC, since the benefits of medical therapy were lower in patients identified and treated in advanced stages of the disease. Moreover, early diagnosis was shown to be useful in recognizing patients with recent onset of heart failure who are not responders to aggressive medical treatments and urgently need heart transplant. PMID- 10687109 TI - [Cardiovascular risk factors in Italy: an interpretation with reference to th National Health Plan 1998-2000. Research Group of the Cardiovascular Epidemiologic Observatory]. AB - The data submitted were collected throughout 1998 in 34 cardiology centers or divisions within the Cardiovascular Epimediological Observatory (Osservatorio Epidemiologico Cardiovascolare), in a general population sample of 3222 men and 3192 women aged 35 to 74 years. Each center screened 200 individuals (100 men and 100 women) randomly selected from the electoral rolls. The data constitute an updated reference benchmark for the health status of the Italian population at the end of the Nineties. It also represents a useful source of information for attainment of the objectives pursued by the National Health Plan 1998-2000: a healthier diet, increased physical activity and reduced smoking. The proportion of hypertensives receiving adequate treatment and the high prevalence of individuals with "undesirable" cholesterol levels are problems that should be dealt with following the preventive actions indicated in the National Health Plan for 1998-2000. PMID- 10687110 TI - [Effectiveness of paroxetine in the treatment of refractory vasovagal syncope in young patients]. AB - OBJECTIVES: The aim of the study was to assess whether the well-tolerated serotonin re-uptake inhibitor paroxetine hydrochloride could prevent vasovagal syncope in young patients resistant to or intolerant of previous traditional therapies. BACKGROUND: Serotonergic mechanisms may play a major role in the pathophysiology of neurocardiogenic syncope, and serotonin re-uptake inhibitors have been recently reported to be effective in preventing episodes. METHODS: Forty-one consecutive young patients (13 male and 28 female), aged less than thirty years with recurrent syncope and positive head-up tilt test, and in whom standard therapies with beta-blocking, vagolytic, negative inotropic or mineral corticoid agents were ineffectual, poorly tolerated or contraindicated, randomly received either paroxetine at 20 mg once a day or a placebo. A head-up tilt test was then re-performed after one month of treatment, and the clinical effect was noted over a mean follow-up of 27.1 +/- 6.6 months. RESULTS: The response rates (negative tilt test) after one month of treatment were 57.1 versus 33.3% (p < 0.001) in the paroxetine and placebo groups, respectively. During follow-up, spontaneous syncope was observed in 4 patients (19%) in the paroxetine group and in 12 patients (60%) in the placebo group (p < 0.001). Only one patient (4.8%) asked to be discontinued from the drug for severe recurrent headache. CONCLUSIONS: Paroxetine significantly improved symptoms of young patients with recurrent vasovagal syncope unresponsive to or intolerant of traditional medications and was well tolerated by patients. PMID- 10687111 TI - [Clinical course of pre- and post-natal isolated congenital atrioventricular block diagnosed in utero]. AB - We evaluated the pre- and postnatal outcome of isolated atrioventricular (AV) block detected during fetal life in order to identify factors that may affect the natural history of this lesion and to assess prenatal therapy. Over the past eight years, we consecutively evaluated 10 fetuses with complete AV block. The mean gestational age at diagnosis was 25.3 weeks and the mean heart rate was 57 bpm; two fetuses were hydropic. During pregnancy, one fetus suddenly died, while 6 out of 9 fetuses had a mean reduction in heart rate of 17.8 bpm; 4 patients had heart rate < 50 bpm. Five fetuses developed heart failure, which was severe in 2 cases and mild in 3. The mean gestational age at delivery was 31 weeks. Dexamethasone was administered to the mothers during pregnancy in 4 cases without modification of AV block and/or of heart rate, but in 3 out of 4 fetuses the general condition remained stable in spite of the reduction in heart rate in two of them. Sympathomimetic drugs were employed in 3 cases with an increase in fetal heart rate, but maternal discomfort appeared in two cases. Three newborns died during the first week of life, two of hydrops and one of persistent pulmonary hypertension. Cardiac pacing was performed in 6/9 patients within the first 8 months of life and in 3 within the first 2 days. In conclusion, morbidity and mortality are high when AV block is detected during fetal life. Negative prognostic factors are hydrops and a heart rate < 50 bpm. Pre-term delivery to enable cardiac pacing is probably the therapy of choice if gestational age is > 27-28 weeks. Sympathomimetic drugs are effective but are poorly tolerated by the mothers. Dexamethasone has no effect on AV block and/or heart rate, but may improve clinical tolerance of conduction disturbance. PMID- 10687112 TI - [Clinical characteristics, familial distribution and preliminary genetic data in 9 different families with "Brugada's syndrome"]. AB - Clinical electrocardiographic evaluation and complete non-invasive assessment including nuclear magnetic resonance (NMR) are reported for 7 subjects with cardiac arrest (CA), 6 due to ventricular fibrillation (VF) and 1 to ventricular tachycardia (VT). Two more subjects, one with and one without a family history of non-resuscitated sudden death (NRSD), were included. All 9 subjects showed the typical pattern of the Brugada's syndrome (BS), characterized by incomplete right bundle branch block, ST T elevation in V1 V3. We globally evaluated 64 subjects belonging to the 9 families examined, 5 of whom were identified in Bologna, 3 in Florence and one in Parma. BS is characterized in the experience described in the present paper by a family distribution of the ECG pattern in different members. Furthermore, a family distribution of NRSD, even at a young age, was observed. Electrocardiographic features were consistent with variable degrees and aspects of the intraventricular conduction delay (ICD) and of the ST T elevation pattern. NMR has been performed so far in 23 out of 64 members examined by echo, and was normal in 17/23, with only 6 showing pathological aspects such as mild dilatation of the right ventricle, reduced thickness of the right free wall, isolated dilatation of the right ventricular infundibulum and other minor pathological aspects. Preliminary genetic screening (GS), performed on 20 members of three families, was negative for the typical genetic patterns of right ventricular dysplasia (ARVD). In six families, GS is still ongoing. Genetic screening of sodium channel pathology is in progress in the same families. In conclusion, BS has been documented in the present paper as a hereditary syndrome, both for clinical and ECG aspects, associated with CA due to VF, which required an AICD implantation, at least in symptomatic subjects. There may exist a CONGENITAL form of BS due to pathology of sodium channels, without a demonstrable structural heart disease and an ACQUIRED form of BS secondary to an initial ARVD. From the clinical point of view, a complete evaluation, including serial ECG, pharmacological testing and programmed electrical stimulation of other subjects in the families, may be important in preventing sudden death, mainly in symptomatic subjects who always require an implantable cardioverter defibrillator. PMID- 10687113 TI - Relief of an early pulmonary pathway obstruction with stenting after modified Fontan operation in an adult. AB - The prognosis of Tricuspid Atresia, a rare congenital heart disease, has been changed by surgery. The criteria for Fontan operation have been well established in the literature and adult patients rarely fulfil these criteria; however, in very selected cases Fontan operation can be performed also in adults. A 33 year old woman with tricuspid atresia and previous palliation with classical right Glenn and with left modified Blalock-Taussing, underwent modified Fontan operation because increasing cyanosis and moderate exercise intolerance. Three weeks after operation the patient was readmitted with severe heart failure because of a tight obstruction at the anastomosis between right atrium and main left pulmonary artery. The stenosis was treated with balloon and stenting achieving large pathway. Our experience confirms that after a modified Fontan, if cardiac failure occurs, an immediate full investigation have to be done. PMID- 10687114 TI - Treatment of a native right coronary artery with the PercuSurge Guardwire protection system during PTCA and stenting. AB - Cardiac catheterization in a patient with recent-onset unstable angina demonstrated a suboccluded dominant right coronary artery (RCA), with angiographic evidence of a large thrombus load and a severe focal stenosis of the left anterior descending (LAD) coronary artery. After abciximab, uneventful PTCA and stenting of the LAD was performed. The thrombus containing lesion of RCA was treated with balloon predilatation and stent deployment, and the whole procedure was accomplished with protection of the distal vessel by means of PercuSurge. This device was planned to avoid distal debris migration during percutaneous interventions of saphenous bypass grafts. The system is designed to allow the placement of a temporary occlusion device, a low-profile balloon, distal to the lesion to be treated during the procedure. The occlusive balloon is kept inflated during the treatment of the lesion. Before deflating the balloon and allowing blood to reach the distal vessel, whenever it is necessary, the material proximal to the balloon is aspirated through a monorail catheter. This aspiration removes blood and thrombi proximal to the occlusive balloon from the treated coronary artery. The case we present first reports the application of the device in a large native coronary artery, with an optimal distal flow restoring and no evidence of thrombus embolization. This type of protection of distal coronary vessels towards micro- and macroembolization of thrombi is a promising system of performing safer percutaneous interventions, even in acute ischemic syndromes. PMID- 10687115 TI - Incessant ventricular tachycardia early after acute myocardial infarction: efficacy of radiofrequency catheter ablation but not of optimal coronary revascularization. AB - Incessant ventricular tachycardia is an arrhythmia refractory to conventional antiarrhythmic treatment. We describe the case of 55-year-old man who presented incessant ventricular tachycardia in the early post-acute phase of myocardial infarction. Optimal coronary revascularization was not effective, but radiofrequency catheter ablation was able to eliminate the anatomic substrate and clinical arrhythmic recurrence. PMID- 10687116 TI - Cardiac tamponade: an unusual, fatal complication of infective endocarditis. AB - Infective endocarditis still occurs in Western countries and so far, it has been an important medical problem. The spectrum of infective endocarditis complications may be extremely wide. We report two unusual cases of infective endocarditis complicated with heart rupture and pericardial effusion. In one case, the infective process spread from the aortic valve developing a sinus of Valsalva aneurysm with subsequent aortic perforation. The perforation reached the right auricular epicardial region with subsequent epicardial rupture and hemopericardium. In the other patient, an infective process of the aortic cusps induced the formation of multiple abscesses in the left ventricle and in the right atrium. An annular abscess of the tricuspid valve was found. From the right atrium, an infected fistula spread through the atrial wall and perforated the epicardial surface of the right auricle. Aside from the rare occurrence of these complications in patients affected with infective endocarditis, these cases are of clinical interest because they raise the problem of the need of greater sensitivity to the diagnosis of endocarditis and proper diagnostic approach. PMID- 10687117 TI - [Proposed tools for quality assurance in echocardiography]. PMID- 10687118 TI - [Organization of pediatric cardiology in Italy. Cognitive survey by the Italian Society of Pediatric Cardiology]. PMID- 10687120 TI - [Words and the specialty's pains]. PMID- 10687119 TI - [Cardiac ischemia (excluding AMI) and DRG classification system. Epidemiologic approach]. PMID- 10687121 TI - [Etiologic diagnosis of dilated cardiomyopathy]. PMID- 10687122 TI - [Role, timing and cost-benefit analysis in non-invasive assessment after non complicated myocardial infarction]. PMID- 10687123 TI - [Risk stratification in patients treated with non-cardiac major vascular surgery]. PMID- 10687124 TI - [Neurogenic syncope in the elderly patient]. PMID- 10687125 TI - [The use of endothelin-1 receptor antagonists in cardiovascular diseases]. PMID- 10687126 TI - [What is the role of nitrates in the treatment of heart decompensation? A rationale for their use]. PMID- 10687127 TI - [Orthostatic tachycardia syndrome. Update on etiology, diagnosis and treatment]. PMID- 10687128 TI - [Suboptimal exercise test in chronic heart decompensation: the six-minute walking test]. PMID- 10687129 TI - [Arrhythmic complications in patients with heart decompensation: when and how to treat]. PMID- 10687130 TI - [Risk identification and strategy for the prevention of sudden cardiac death after myocardial infarction]. PMID- 10687131 TI - [Heart rupture in acute myocardial infarction: advantages of the use of Doppler color echocardiography M-2D in coronary intensive care unit]. PMID- 10687132 TI - Cytotoxic T lymphocytes from humans with adenocarcinomas stimulated by native MUC1 mucin and a mucin peptide mutated at a glycosylation site. AB - MUC1 mucin peptides stimulated cytotoxic T lymphocytes (CTL) from humans with adenocarcinomas. Peripheral blood mononuclear cells, tumor-draining lymph node cells, or tumor-infiltrating lymphocytes were stimulated using mono-nuclear cells from humans with adenocarcinomas of breast or ovary, respectively, using (a) a native MUC1 mucin tandem repeat peptide of 20 amino acids (MUC1-mtr1) plus recombinant human interleukin-2 (IL-2), (b) the mutated (T3N) MUC1-mtr1 plus IL 2, or (c) immobilized anti-CD3 plus IL-2, or (d) IL-2 alone. The CTL stimulated by each of these four conditions were predominately CD4+. However, the CTL stimulated by either the native MUC1-mtr1 or (T3N) MUC1-mtr1 showed 5-10 times greater cytotoxicity of a breast cancer cell line that expresses MUC1 compared to CTL stimulated by either anti-CD3 + IL-2 or IL-2 alone. Each incubation condition generated CTL with different variable beta gene families of T-cell receptors, implying an oligoclonal expansion of a limited CTL repertoire for each. Thus, peptide-stimulated T cells showed expression of cytotoxic cells, which was not induced by nonspecific (anti-CD3 or IL-2) stimulation. PMID- 10687133 TI - Interleukin-4 enhances the in vitro precursor cell recruitment for tumor-specific T lymphocytes in patients with glioblastoma. AB - Previously the authors showed that interleukin-4 (IL-4), used in combination with IL-2, increases the reduced proliferation rate of T cells of glioblastoma-bearing patients after in vitro autologous immunization. In this report, they sought to determine whether this effect is caused by a direct mitogenic effect of IL-4, or rather by an indirect effect through an increased expression of the IL-2 receptor subunits or an enhanced recruitment of responsive cells. Flow cytometric analysis confirmed that the IL-2 receptor subunits are less expressed on circulating T cells from patients with glioblastoma than on those from healthy donors. Because no significant modification of the expression of the p55 and p75 subunits of the IL-2 receptor is observed in cultures treated with both IL-2 and IL-4, the reported enhanced proliferation rate cannot be attributed to an increased level of IL-2 receptor expression. Limiting dilution assays, using autologous target cell immunization, show that treatment with both cytokines (IL-2 plus IL-4) significantly increases the number of recruitable precursor cells without affecting their proliferation rate. These results indicate that IL-4 facilitates an immune response against the autologous tumor cells in glioblastoma-bearing patients by increasing the recruitable precursor T-cell frequency. PMID- 10687134 TI - Recognition of shared melanoma antigens in association with major HLA-A alleles by tumor infiltrating T lymphocytes from 123 patients with melanoma. AB - A total of 123 tumor-infiltrating T lymphocyte (TIL) cultures established from patients with HLA-A1, -A2, -A3, -A24, or -A31 metastatic melanoma in the Surgery Branch, National Cancer Institute, were screened for recognition of shared melanoma antigens including five melanosomal proteins (tyrosinase, MART-1/melan A, gp100, TRP1, TRP2) as well as peptides derived from MAGE-1 and MAGE-3. Examination of the specificity of these T cells indicated that 16% of HLA-A1 TIL, 57% of HLA-A2 TIL, 7% of HLA-A3 TIL, 13% of HLA-A24 TIL, and 27% of HLA-A31 TIL recognized shared melanoma antigens restricted by major histocompatibility complex class I. Melanosomal proteins were frequently recognized by these TIL, and MART-1(27-35), gp100(154-162), gp100(209-217), and gp100(280-288) represent highly immunogenic epitopes that were recognized by a high percentage of HLA-A2 restricted melanoma reactive TIL. Recognition of gp100 by HLA-A2 restricted TIL significantly correlated with clinical response to adoptive immunotherapy with TIL in 21 HLA-A2 melanoma patients (p = 0.024). Four HLA-A1, two HLA-A2, two HLA A3, one HLA-A24, and two HLA-A31 restricted shared antigen-specific TIL did not recognize the previously identified antigens tested in this study, and may be useful for the identification of new melanoma antigens. The observation that TILs isolated from patients with metastatic melanoma recognized melanosomal proteins in the context of predominant HLA-A alleles implies that it may be possible to develop immunotherapies for patients with melanoma expressing diverse HLA types. PMID- 10687135 TI - Reduced recognition of metastatic melanoma cells by autologous MART-1 specific CTL: relationship to TAP expression. AB - Class I expression in context with T-cell receptor expression is crucial for peptide presentation and induction of CD8+ cytotoxic T lymphocytes (CTL). Presentation of class I bound peptides is dependent on transporter-associated proteins (TAP) expression and function. Tumor infiltrating lymphocytes from a patient with melanoma were isolated, expanded in vitro in the presence of interleukin-2, and tested for cytotoxicity against HLA-A2 positive, MART-1 positive autologous tumor cells, an HLA-A2-positive, MART-1 positive melanoma cell line (Mel-501), and HLA-A2-negative melanoma cells. Significant killing occurred against both A2-positive cell lines (63% and 65%, respectively), but not against the A2-negative line (18%) or A2-positive autologous tumor (1.5%). These CTL preferentially recognized the MART-1 peptide F119, 27-35, and gp100 peptide F125, 280-288, resulting in a 30% to 60% enhancement of lysis when autologous tumor or major histocompatibility complex class I "empty" T2 cells were pulsed with either peptide. To address whether the deficiency in autologous tumor recognition might be related to a deficiency in Ag presentation, we screened for the presence of TAP1 and TAP2 transcripts by polymerase chain reaction, Southern blotting, and scanning densitometry using sequence-specific primers and probes. Both TAP1 and TAP2 expression levels in the autologous tumor were minimal, yet were upregulated 7- to 18-fold, respectively, by interferon-gamma. Despite this increase, a similar increase in cytotoxicity did not occur. In short, deficiencies in TAP presentation may have functional significance for tumor escape from immunosurveillance and with respect to impending vaccine trials. PMID- 10687136 TI - Tumorigenicity and immunogenicity of murine tumor cells expressing an MHC class II molecule with a covalently bound antigenic peptide. AB - The significance of CD4+ lymphocytes and major histocompatibility complex (MHC) class II-restricted antigens in antitumor immunity has been demonstrated in several animal models as well as in some human tumors. However, because of the lack of known class II-restricted antigens, the participation of CD4+ cells in antitumor responses has not been well characterized. Recent reports showed that class II proteins covalently linked to an antigenic peptide could be constructed and cells expressing these fusion proteins were recognized by specific TH cells. The aim of this study was to determine the effect of the expression of a class II peptide construct on the tumorigenicity and immunogenicity of transfected murine tumor cells. We have constructed a gene for I-Ed beta chain covalently coupled to the I-Ed-restricted TH cell determinant of sperm whale myoglobin (SWM132-145). This class II fusion protein was recognized by a specific TH cell line on the surface of COS-7 cells or BALB/c sarcoma cells. The sarcoma cells expressing the MHC-peptide complex were rejected by immunocompetent BALB/c mice, and in vivo T cell subset depletion experiments suggested the importance of CD4+ cells in the rejection. Moreover, splenocytes from mice immunized with tumor cells expressing the I-Ed-SWM complex showed specific peptide recognition in vitro. Such covalent MHC-peptide complexes could prove useful in studies on the role of CD4+ lymphocytes in antitumor immune responses, and also in designing new, more effective vaccine approaches to the immunotherapy of cancer, as class II restricted tumor-associated antigens are identified for human cancers. PMID- 10687137 TI - Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL-4, and TNF-alpha are functional antigen-presenting cells resembling mature monocyte-derived dendritic cells. AB - Dendritic cells (DCs) are powerful antigen-presenting cells. Because DCs are rare cells, methods to produce them in vitro are valuable ways to study their biologic properties and to generate cells for immunotherapy. This study defines the antigen-presenting properties of DCs generated in vitro from CD34+ cells of patients with breast cancer. The combination of cytokines flt3 ligand + c-kit ligand + granulocyte-macrophage colony-stimulating factor (GM-CSF) + interleukin 4 (IL-4) + tumor necrosis factor-alpha (TNF-alpha) was used to maximize the output of mature DCs in the culture of CD34+ cells while minimizing the production of monocytes. Cells grew and differentiated into DCs as measured by a time-dependent upregulation of cell surface antigens major histocompatibility complex class II, CD1a, CD80, CD86, CD40, and CD4, so that 40% +/- 9% (n = 6) of cells in culture at day 15 were CD1a+CD14-. Markers were acquired in the same sequence as on monocytes induced to differentiate with GM-CSF + IL-4. Differentiation was marked by a time-dependent increase in allostimulatory function, which, at its peak, was more potent than in cultures of DCs generated from monocytes with GM-CSF + IL-4, but was comparable on a cell-to-cell basis to that of mature monocytes cultured in flt3-ligand + c-kit-ligand + GM-CSF + IL-4 + TNF-alpha. Both CD34+ cell-derived and monocyte-derived DCs were able to process and to present tetanus toxoid and keyhole limpet hemocyanin to autologous T cells and to present major histocompatibility class I-binding peptides to CD8+ cytotoxic T lymphocytes inducing interferon-gamma production. Altogether, these results suggest that DCs generated from CD34+ cells of patients with breast cancer with flt3 ligand, c-kit ligand, GM-CSF, IL-4, and TNF-alpha are competent antigen-presenting cells, particularly for CD8+ cytotoxic T lymphocytes, and resemble mature monocyte-derived DCs in the assays described here. PMID- 10687138 TI - Generation of T-cell immunity to a murine melanoma using MART-1-engineered dendritic cells. AB - The murine melanoma B16 expresses the murine counterpart of the human MART 1/Melan-A (MART-1) antigen, sharing a 68.6% amino acid sequence identity. In this study, mice were vaccinated with bone marrow-derived murine dendritic cells genetically modified with a replication-incompetent adenoviral vector to express the human MART-1 gene (AdVMART1). This treatment generated a protective response to a lethal tumor challenge of unmodified murine B16 melanoma cells. The response was mediated by major histocompatibility complex class I-restricted cytotoxic T lymphocytes specific for MART-1 antigen, which produced high levels of interferon gamma when reexposed to MART-1 in vitro and lysed targets in a calcium-dependent mechanism suggestive of perforin/granzyme B lysis. MART-1 was presented by the dendritic cells used for vaccination and not by epitopes cross-presented by host antigen-presenting cells. In conclusion, dendritic cells genetically modified to express the human MART-1 antigen generate potent murine MART-1-specific protective responses to B16 melanoma. PMID- 10687139 TI - Cervical cancer cells induce apoptosis of cytotoxic T lymphocytes. AB - The goal of immunotherapy is to eliminate tumors by generating tumor-specific cytotoxic T lymphocytes (CTLs) in patients or by adoptively transferring ex vivo activated CTLs into patients. Clinical trials have shown that tumor-specific CTLs often disappear before tumors are completely eliminated. In this study, the authors show that CTLs specific for cervical tumor cells undergo apoptosis after they are co-cultured with cervical tumor cells. The established cervical tumor cell lines and cervical cancer tissues express CD95 (Fas/Apo-1) ligand. The tumor cell-induced T-cell apoptosis can be blocked by an inhibitory anti-CD95 (APO 1/Fas) antibody, indicating that tumor cells induce apoptosis of CTLs through CD95-CD95 ligand interaction. Addition of interleukin-2 (IL-2) and IL-7 into the culture rescues the CTL from tumor cell-induced apoptosis. The rescued T cells retain their full antitumor cytotoxicity. These data suggest that human cervical tumor cells might actively down-regulate a cellular immune response by inducing apoptosis of specific T cells during immunotherapy. Local use of IL-2 and IL-7 as adjuvants may promote survival of the CTL and, thus, enhance the efficacy of immunotherapy. PMID- 10687140 TI - Enhanced transgene expression and effective in vivo antitumor immune responses initiated by dendritic progenitors transfected with a nonviral T7 vector expressing a model tumor antigen. AB - Genetic education of dendritic cells (DCs) with tumor-associated antigens is an encouraging development in DC-mediated tumor immunotherapy. In this study, to increase the transgene expression by DCs using nonviral vectors, a cytoplasmic T7 vector (T7T7/T7Luc) was used to transfect bone marrow-derived DCs with the firefly luciferase gene as a reporter and as a model tumor antigen. As a result, the luciferase activity of T7T7/T7Luc-transfected DCs was more than four times greater than that of DCs transfected with pCMVLuc, a commonly used nonviral vector. Furthermore, the luciferase activity was increased three times more when dendritic progenitor cells rather than mature DCs were transfected. In vivo tumor studies showed that T7T7/T7Luc-transfected DCs, which express high levels of luciferase (model tumor antigen), stimulated a stronger immune response than did pCMVLuc-transfected DCs, which express relatively low levels of luciferase, as indicated by the cytotoxic T lymphocyte assay. T7T7/T7Luc transfected DCs, when injected into recipient mice, evoked an antigen-specific immune response that can effectively eradicate implanted metastasis and prevent new tumor development by murine melanoma cells genetically modified to express luciferase. Therefore, the T7 system is a powerful nonviral vector that can be used to genetically educate DCs with tumor-associated antigens for tumor immunotherapy. PMID- 10687141 TI - Immunomodulatory dendritic cells generated from nonfractionated bulk peripheral blood mononuclear cell cultures induce growth of cytotoxic T cells against renal cell carcinoma. AB - Dendritic cells (DCs) loaded with tumor antigens have the potential to become a powerful tool for clinical cancer treatment. Recently, the authors showed that a tumor-specific immune response can be elicited in culture via stimulation with autologous renal tumor lysate (Tuly)-loaded DCs that were generated from cytokine cultured adherent peripheral blood mononuclear cells (PBMCs). Here, the authors show that immunomodulatory DCs can be generated directly from nonfractionated bulk PBMC cultures. Kinetic studies of DC differentiation and maturation in PBMC cultures were performed by monitoring the acquisition of DC-associated molecules using fluorescence-activated cell sorting analysis to determine the percentage of positive immunostained cells and the mean relative linear fluorescence intensity (MRLFI). Compared with conventional adherent CD14+ cultures, which have mostly natural killer, T, and B cells removed before cytokine culture, bulk PBMC cultures exhibited an early loss of CD14+ cells (day 0 = 78.8%, day 2 = 29.6% versus day 0 = 74%, day 2 = 75%) with an increase in yield of mature DCs (DC19- CD83+) (day 5 = 17%, day 6 = 21%, day 7 = 22% versus day 5 = 11%, day 6 = 15%, day 7 = 23%). Although a comparable percentage of DCs expressing CD86+ (B7-2), CD40+, and HLA-DR+ were detected in both cultures, higher expression levels were detected in DCs derived from bulk culture (CD86 = MRLFI 3665.1 versus 2662.1 on day 6; CD40 = MRLFI 1786 versus 681.2 on day 6; HLA-DR = MRLFI 6018.2 versus 3444.9 on day 2). Cytokines involved in DC maturation were determined by polymerase chain reaction demonstrating interleukin-6 (IL-6), IL-12, interferon gamma, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor-alpha mRNA expression by bulk culture cells during the entire 9-day culture period. This same cytokine mRNA profile was not found in the conventional adherent DC culture. Autologous renal Tuly (30 micrograms protein/10(7) PBMCs) enhanced human leukocyte antigen expression by DCs (class I = 7367.6 versus 4085.4 MRFLI; class II = 8277.2 versus 6175.7 MRFLI) and upregulated cytokine mRNAs levels. Concurrently, CD3+ CD56-, CD3+ CD25+, and CD3+ TCR+ cell populations increased and cytotoxicity against autologous renal cell carcinoma tumor target was induced. Specific cytotoxicity was augmented when cultures were boosted continuously with IL-2 (20 U/mL biological response modifier program) plus Tuly stimulation. These results suggest that nonadherent PBMCs may participate in enhancing DC maturation. Besides the simplicity of this culture technique, bulk DC cultures potentially may be used with the same efficiency as conventional purified DCs. Furthermore, bulk culture-derived DCs may be used directly in vivo as a tumor vaccine, or for further ex vivo expansion of co cultured cytotoxic T cells to be used for adoptive immunotherapy. PMID- 10687142 TI - Cytokine-inducing activity and antitumor effect of a liposome-incorporated interferon-gamma-inducing molecule derived from OK-432, a streptococcal preparation. AB - An interferon-gamma (IFN-gamma)-inducing molecule (OK-PSA) has been purified from OK-432 by an affinity chromatographic technique performed on cyanogen bromide activated Sepharose 4B-bound TS-2 monoclonal antibody, which neutralizes IFN gamma-inducing activity of OK-432. OK-PSA has striking anti-tumor activity in vivo and in vitro. In the current study, the liposomes were used to improve the delivery of the agent (OK-PSA) to effector cells and to increase the therapeutic effect. Significantly less OK-PSA encapsulated into liposomes (Lipo-OK-PSA) than OK-PSA alone (1/100 or less of OK-PSA alone) was required to induce IFN-gamma, tumor necrosis factor-alpha (TNF-alpha), TNF-beta, interleukin-1 beta (IL-1 beta), natural killer, and lymphokine-activated killer activities by human peripheral blood mononuclear cells and mouse spleen cells. Furthermore, higher levels of these activities were detected in peripheral blood mononuclear cells and mouse spleen cells treated with Lipo-OK-PSA than in those treated with OK PSA. All of these activities induced by Lipo-OK-PSA were almost completely neutralized by anti-asialo-GM1 antibody and complement (p < 0.001). In in vivo experiments, Lipo-OK-PSA elicited striking anti-tumor activity on syngeneic Meth A tumor-bearing and colon 26-bearing BALB/c mice and on salivary gland tumor bearing nude mice far better than did OK-PSA. Furthermore, high levels of natural killer and lymphokine-activated killer activities and a significant increase in the number of cells positive for asialo-GM1, IFN-gamma, TNF-alpha, or IL-1 beta were detected in the spleen cells derived from the animals given Lipo-OK-PSA compared with those given saline. These findings clearly indicate that OK-PSA plays an important role in the anti-tumor efficiency of OK-432, and that, for the most part, liposome encapsulation of this molecule markedly accelerates its effect mediated by asialo-GM1-positive cells (mainly natural killer cells). PMID- 10687143 TI - c-erbB-2 and episialin challenge host immune response by HLA class I expression in human non-small-cell lung cancer. AB - The role of major histocompatibility complex expression in cancer prognosis and pathogenesis is contradictory. The aim of the current study was to compare the expression of HLA class I molecules and of oncoproteins that may be sources of peptides presented by HLA class I antigens in non-small-cell lung cancer. For this purpose, the expression of HLA class I antigen and TAP-1 molecule (a transporter in the antigen-processing 1 transport protein) were studied with epidermal growth factor, receptor; c-erbB-2; episialin; wild-type and mutant p53; bcl-2 oncoprotein expression; and angiogenic factor expression (vascular endothelial growth factor and thymidine phosphorylase). The degree of lymphocytic stromal infiltration and of platelet-endothelial cell adhesion molecule expressing lymphocytes was also studied. A strong association of c-erbB-2 and MUC1 (episialin) expression with HLA class I expression was observed (p = 0.005 and 0.009, respectively). Intense CD31-positive lymphocytic infiltration was also more frequent in HLA class I-positive cases (p = 0.05). Although there was no association of HLA class I expression with survival, loss of the HLA class I expression in MUC1 or c-erbB-2 overexpressing cases conferred a poorer clinical outcome (p = 0.04). Both c-erbB-2 and MUC1 are well-known targets of T-cell mediated cytotoxicity and cell-cell or cell-matrix adhesion-regulating proteins. The authors provide evidence that the sequence of cell adhesion-disrupting oncoprotein expression, HLA class I induction, and enhanced epitope presentation followed by lymphocytic response is an important pathogenetic three-step sequence of events that define, in part, the clinical outcome in non-small-cell lung cancer. PMID- 10687144 TI - Vitamin D3 treatment to diminish the levels of immune suppressive CD34+ cells increases the effectiveness of adoptive immunotherapy. AB - Tumor growth can increase the number of immature bone marrow-derived CD34+ cells that exhibit natural suppressor (NS) activity toward T-cell function. Using a metastatic Lewis lung carcinoma (LLC-LN7) tumor model, these CD34+ NS cells were shown to be present within the s.c. primary tumor tissue, but their levels declined after treatment with the inducer of myeloid cell differentiation, vitamin D3. Therefore, studies determined whether vitamin D3 treatment to diminish the CD34+ NS cell levels in LLC-LN7-bearing mice would enhance (a) intratumoral immune reactivity and (b) the antitumor activity of adoptive therapy consisting of tumor-reactive lymph node cells. The results showed that vitamin D3 treatment alone increased the intratumoral CD8+ cell content and the activity of the intratumoral infiltrate, as detected by production of interferon-gamma and expression of the p55 IL-2 receptor. Although vitamin D3 treatment had no effect on the size of the primary tumor, it lessened the extent of tumor metastasis. Treating mice with the combination of vitamin D3 and adoptive immunotherapy significantly reduced metastasis in mice with established tumors, and reduced both metastasis and locoregional recurrence after surgical excision of the primary tumor. These studies demonstrate that vitamin D3 treatment increases intratumoral T-cell immune reactivity, and that coupling vitamin D3 treatment to diminish levels of CD34+ NS cells with adoptive immunotherapy enhances the effectiveness of the adoptively transferred tumor-reactive lymph node cells at limiting both metastasis and locoregional tumor recurrence. PMID- 10687145 TI - Antitumor vaccination using a major histocompatibility complex (MHC) class I restricted pseudopeptide with reduced peptide bond. AB - Synthetic peptides have raised a considerable interest in the fields of vaccines and immunotherapy. The authors previously introduced modifications into the peptide backbone of the H-2Kd-restricted epitope CW3. One of these pseudopeptides, C7, bound to Kd with an affinity identical to the parent peptide and was recognized by T cells specific for the parent peptide. The authors now show that this analog has an increased resistance to trypsin and displays an extended half-life in serum. The authors further tested its immunogenicity both in vitro and in vivo and found that cytotoxic T lymphocytes (CTL) induced against the peptide analog recognize the parent peptide. Moreover, analysis of T-cell receptor rearrangements by Immunoscope software revealed that C7-induced CTL display the hallmarks of the response against the parental epitope CW3. Administration of the pseudopeptide into DBA/2 mice induces a protective immune response against a lethal challenge with tumor cells expressing the parent peptide. Therefore, modifications in the backbone of antigenic peptides can decrease protease susceptibility while preserving immunogenicity. Such peptide analogues may therefore prove useful for the development of new therapeutic tools aimed at eradicating pathogens or tumors. PMID- 10687146 TI - Delivery of cytokines by liposomes: hematopoietic and immunomodulatory activity of interleukin-2 encapsulated in conventional liposomes and in long-circulating liposomes. AB - Although liposomal delivery of interleukin-2 (IL-2) and other cytokines improves their pharmacokinetics and biologic activity in vivo, there are no comparative functional studies of various liposomal formulations as cytokine carriers. In the present investigation, recombinant human IL-2 was encapsulated in two formulations of large (mean diameter 0.75-1.5 microns) multilamellar vesicles (MLV, referred to as conventional liposomes) or in small (mean diameter, 60 nm), unilamellar, long-circulating liposomes (referred to as sterically stabilized liposomes, SSL). The biologic activity of the liposomal formulations and of free IL-2 was tested in parallel in vitro and in mice. The main observations were as follows: (a) All the liposomal IL-2 (Lip-IL-2) formulations were more efficient than soluble IL-2 in stimulating spleen cell proliferation and lymphokine activated killer (LAK) cell activation in vitro, particularly at low cytokine doses (1-100 CU/mL). (b) After i.v. injection, the circulation time of MLV-IL-2 and SSL-IL-2 was 7 and 17 times greater, respectively, than that of soluble IL-2. (c) In comparison with IL-2, all Lip-IL-2 formulations caused a marked increase in the leukocyte levels in blood, spleen, and peritoneal exudate, especially in those of myeloid origin (neutrophils, eosinophils, immature granulocytes, and macrophages). (d) Although SSL-IL-2 exhibited the longest circulation time, MLV IL-2 was more potent in elevating leukocyte levels and in triggering LAK cell activity in vivo. (e) The route of Lip-IL-2 administration greatly affected the immunomodulatory activity in the various compartments. (f) MLV-IL-2 proved to be a much more efficient immunoadjuvant than free IL-2 for influenza subunit vaccines as well as for tumor cell vaccines. These findings lend support to our previous studies in which we demonstrated the superior immunomodulatory activity of liposomal IL-2, and suggest that cytokine pharmacokinetics, biodistribution, and pharmacodynamics are markedly influence both by liposomal formulation and route of administration. PMID- 10687147 TI - Phase I study of single, escalating doses of a superantigen-antibody fusion protein (PNU-214565) in patients with advanced colorectal or pancreatic carcinoma. AB - To develop a T-cell-based therapy for carcinomas, the superantigen staphylococcal enterotoxin A (SEA) was supplied with tumor specificity by means of a recombinant fusion of the Fab fragment of the monoclonal antibody C242 recognizing human colorectal (CRC) and pancreatic carcinomas (PC). Using this Fab-SEA fusion protein (PNU-214565), potent cytotoxicity by activation of T cells can be obtained in the targeted area. Twenty-one patients with CRC and 3 with PC were treated with single, escalating doses of PNU-214565 to establish the maximum tolerated dose (MTD) and to define toxicities. The doses ranged from 0.01 ng/kg to 4.0 ng/kg with three patients at each dose level, except for the dose of 1.5 ng/kg with which six patients were treated because of dose-limiting toxicity. Adverse events (AE) were transient: 13 patients experienced mild to moderate fever. In one patient, a grade 3 fever was followed by a grade 2 hypotension. Other mild or moderate AEs were fatigue, nausea, vomiting, diarrhea, and abdominal pain. No significant hematological toxicity occurred. Immune activation was highly variable with strong activity in peripheral blood seen only in two patients at the dosage level 1.5 ng/kg. They showed pronounced elevations of interleukin-2 (IL-2), IL-6, tumor necrosis factor-alpha, and interferon-gamma, 3 5 hours after the start of infusion. In one patient, IL-2 and IL-6 increased substantially (2,925 U/mL and 32,000 U/mL) concomitantly with grade 3 fever and transient grade 2 neutropenia, grade 2 lymphopenia, and grade 2 monocytopenia. In conclusion, a single 3-hour infusion of PNU-214565 could be safely administered up to 4 ng/kg. MTD was not determined. Instead, a repeat-dose trial was initiated starting at 0.5 ng/kg, considered safe in this trial, with the objective of defining the MTD. PMID- 10687148 TI - Posttransplant adoptive immunotherapy with activated natural killer cells in patients with metastatic breast cancer. AB - Relapse after high-dose chemotherapy is the main cause of therapeutic failure in patients with metastatic breast cancer. Adoptive immunotherapy with interleukin-2 (IL-2) plus activated natural killer cells may eliminate residual disease without excessive toxicity. The authors sought to determine if immunotherapy immediately after transplantation would affect engraftment and the toxicity associated with transplantation. Fifteen consecutive patients with metastatic breast cancer were allocated to three cohorts. Cohort 1 (five patients) received high-dose cyclophosphamide, thiotepa, and carboplatin (CTCb) followed by peripheral blood stem cell infusion and granulocyte colony-stimulating factor at 10 micrograms/kg. Cohort 2 (five patients) received in addition rhIL-2 (2 x 10(6) IU/m2/day) for 4 days intravenously via continuous infusion after peripheral blood stem cell infusion. In cohort 3 (five patients), peripheral blood stem cell transplant was followed by infusion of autologous activated NK cells and rhIL-2 (2 x 10(6) IU/m2/day) for 4 days (via continuous intravenous infusion). Generation of activated NK cells was possible in all patients in cohort 3. All patients has successful engraftment. Median time to absolute neutrophil count more than 0.5 x 10(9)/L was 8 days (range, 8 to 11 days) in cohort 1, 9 days (range, 7 to 11 days) in cohort 2, and 9 days (range, 8 to 9 days) in cohort 3. Median time until the platelet count was more than 20 x 10(9)/L was 14 days (range, 9 to 22 days) in cohort 1, 11 days (range, 6 to 14 days) in cohort 2, and 12 days (range, 11 to 21 days) in cohort 3. All patients developed neutropenic fevers, but the overall toxicity associated with the infusion of IL-2 (cohort 2) or IL-2 plus activated NK cells (cohort 3) did not differ from that observed in cohort 1. Complete responses were achieved in one patient in cohort 1, in two patients in cohort 2, and in one patient in cohort 3. In conclusion, post-transplant adoptive immunotherapy with activated NK cells plus IL-2 is feasible, well tolerated, and does not adversely affect engraftment. PMID- 10687149 TI - Phase II study of combined immunotherapy, chemotherapy, and radiotherapy in the postoperative treatment of advanced non-small-cell lung cancer. AB - The association of adoptive immunotherapy (AI) and radiotherapy has been shown to be effective in the control of residual intrathoracic disease, while having no systemic advantages, in patients operated on for locally advanced non-small-cell lung cancer (NSCLC). The potential synergy of coupling immunotherapy and chemotherapy has been emphasized in several tumors including NSCLC. The aim of this work was to determine the feasibility and activity of a combined therapeutic program, including AI, chemotherapy, and radiotherapy in patients who had undergone incomplete resections for NSCLC. In a phase II trial, 13 patients received the combined treatment. AI was given from week 4 after surgery until week 8. Concurrent chemo-(cisplatin and etoposide)-radiotherapy (60 Gy) was given from week 9 to week 14. Twenty eligible patients received chemoradiotherapy only and were used as a non-randomized concomitant group for merely descriptive purposes. At 9-month follow-up, 10 of the 13 patients had progression of disease and the study was stopped. Progression-free survival and survival were similar to those of the chemoradiotherapy group. The present study showed that the sequence of immunotherapy followed by chemotherapy is not effective as adjuvant treatment in patients operated on for stage III NSCLC, at least when used according to the adopted schedule. PMID- 10687151 TI - Microwave facilities for welding thermoplastic composites and preliminary results. AB - The wide range of applications of microwave technology in manufacturing industries has been well documented (NRC, 1994; Thuery, 1992). In this paper, a new way of joining fibre reinforced thermoplastic composites with or without primers is presented. The microwave facility used is also discussed. The effect of power input and cycle time on the heat affected zone (HAZ) is detailed together with the underlying principles of test piece material interactions with the electromagnetic field. The process of autogenous joining of 33% by weight of random glass fibre reinforced Nylon 66, polystyrene (PS) and low density polyethylene (LDPE) as well as 23.3% by weight of carbon fibre reinforced PS thermoplastic composites is discussed together with developments using filler materials, or primers in the heterogenous joining mode. The weldability dependence on the dielectric loss tangent of these materials at elevated temperatures is also described. PMID- 10687150 TI - Dendritic cells loaded with MART-1 peptide or infected with adenoviral construct are functionally equivalent in the induction of tumor-specific cytotoxic T lymphocyte responses in patients with melanoma. AB - Immunization with tumor-specific-associated antigen--pulsed dendritic cells has proved to be efficacious in various animal models and is being evaluated for the treatment of cancer in humans. Use of dendritic cells pulsed with specific peptides or transfected with tumor-associated antigen genes has been a focused area of investigation for inducing potent tumor and viral immune responses. In this study, the authors demonstrate transgene expression, including the lacZ and MART-1 genes, in dendritic cells infected with adenoviral constructs. These transiently transduced dendritic cells, derived from melanoma patients' monocytes cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4, express the transgene and can stimulate patients' CD8+ T cells to elicit an antitumor immune response comparable to dendritic cells loaded with a defined peptide. These cytotoxic T lymphocytes were able to recognize both known and unknown tumor-associated antigen epitopes and exhibited cytolytic activity against HLA-matched tumor cells expressing the antigen. The ability to induce tumor-specific cytotoxic T lymphocytes in vitro using gene-modified dendritic cells that transiently express tumor-associated antigens demonstrates the potential use of these antigen-presenting cells for developing in vivo cancer vaccines. PMID- 10687152 TI - Switch mode power supply for microwave heating based on the Boucherot effect. AB - This paper describes a new self-resonating switch-mode power-supply for driving CW magnetrons, based on the Boucherot effect. A detailed circuit analysis is given and its performance is evaluated for an 800 W/2450 MHz magnetron, whilst work at high power driving a magnetron up to 40 kW is reported. A comparison of the supply with the conventional power-supply used in microwave ovens is made and the principal features of the new design are found to be: low energy dissipation under short-circuit conditions, low ripple current and voltage waveforms that result in more precise control in the range 20-100% of rated power, high efficiencies and small size and weight. PMID- 10687153 TI - Microwave enhanced ion exchange of cationic and anionic clays. AB - The ion exchange reactions of both cationic clays (e.g. montmorillonites) and of layered double hydroxides ("anionic clays") are greatly accelerated when carried out under conditions of microwave heating. The observations appear general, and the new methodology should be of considerable importance in many aspects of clay science. PMID- 10687154 TI - Temperature and weight loss profiles of model cakes baked in the microwave oven. AB - Model cake systems were formulated with wheat and rice starches at hydration levels of 112.5% and 137.5% (flour weight basis) and baked in a microwave oven at power levels of 80% and 100%. Temperature profiles and weight loss profiles of the cakes baked in the microwave oven were compared with those of the cakes baked conventionally. One cake was baked at a time, and three replications of each treatment were used. Center and edge temperatures of microwave cakes increased significantly with increasing moisture content of the batter and oven power. Weight loss of the cakes was dependent on oven power, starch type and hydration levels. Cakes baked in the microwave oven had greater weight loss than the cakes baked in convection mode. Wheat starch cakes had greater volumes than rice starch cakes. Rice starch cakes were more tender than wheat starch cakes. PMID- 10687155 TI - Variable frequency microwave heating of food. AB - Industrial microwave food processing is universally based on single frequency microwave sources. With the emergence of variable frequency microwave ovens, it is possible to exploit the frequency dependence of a food's permittivity and/or choice of heating frequency, for example as a new route to achieving targeted heating. Variable frequency heating procedures are developed to overcome the geometry of a roughly spherical foodstuff dominating the heating pattern when heated in fixed frequency applicators. Target mean temperatures of 55, 75 and 90 degrees C within 2 minutes and without physical damage were set; means of 54.5 +/ 4.1, 75.1 +/- 4.7 and 87.6 +/- 3.5 degrees C respectively were achieved within the time constraint and with no major physical damage, based on combining 8 discrete frequencies between 2.4 and 6.2 GHz. PMID- 10687156 TI - Enzyme inactivation analyses for industrial blanching applications employing 2450 Mhz monomode microwave cavities. AB - Browning reactions in fruits and vegetables are recognized as a serious problem for the European food industry, particularly for the mushroom sector. The major enzyme responsible for the browning reaction is polyphenoloxidase (PPO). In this paper considerable reduction has been achieved in both the time and temperature required for complete microwave enzyme inactivation compared to conventional hot water treatments, which can be translated into both increased benefits and enhanced quality products for the food industry. Furthermore, the short exposure time required for complete inactivation of aqueous solutions of PPO irradiated with microwaves within monomode cavities is very important to reduce the browning rate of mushroom extracts, and could lead to a much greater product profitability when treating whole processed mushrooms. PMID- 10687157 TI - Exercise test in detecting anomalous behaviour of blood pressure in patients successfully operated on for coarctation of the aorta. AB - Twenty-eight patients operated with success for isolated coarctation of the aorta (i.e. with normal blood pressure and upper/lower limb gradient < or = 20 mmHg at rest) underwent exercise testing to evaluate blood pressure and upper/lower limb pressure gradient during exercise. At maximum effort: 57% (16/28) of the patients were hypertensive and 43% (12/28) of patients increased upper/lower limb gradient over 35 mmHg. No significant correlation was found between the age at surgery (before or after 3 years of age) and maximal systolic blood pressure on exercise and maximal pressure gradient on exercise. The 12 patients with an exercise pressure gradient > 35 mmHg underwent digital angiography or magnetic resonance of the aorta. In 7 cases a mild residual narrowing was found (5 with mild transverse aortic arch hypoplasia, 2 with a mild residual coarctation). In 5 cases no residual narrowing was present. Many factors are thought to be involved in the anomalous behaviour of blood pressure during effort: in some cases anatomic factors, as residual narrowings of the aorta, in other cases functional factors, as increased forearm vascular reactivity, altered baroceptor function, different reactivity and structure of the pre- and post-stenotic aorta, etc., in other cases finally, both factors, anatomic and functional. We conclude that the exercise testing provides the best information on blood pressure modifications during activity and it allows to us to identify those patients which, because of exercise-induced hypertension, remain at risk of premature cardiovascular disease, also after a successful operation. However, when hypertension is found during effort, a strenous physical activity should be avoided and antihypertensive treatment may be required. So the cardiovascular risk due to hypertension can be reduced in the long term follow-up. Corrective surgery for coarctation of the aorta, introduced in 1944, has completely modified the natural history of the disease. Nowadays the operative risk is very low for isolated coarctation and the great majority of the patients is asymptomatic after surgical repair. Nevertheless, their life expectancy is substantially shortened, if compared with the survival curve of a normal population. The vascular and cardiovascular accidents, usually related to residual systemic hypertension, are the most common cause of this. Some studies in the literature have shown that many patients with normal blood pressure and no or little residual upper/lower limb pressure gradient at rest, may develop an anomalous blood pressure response e and/or a high upper/lower limb pressure gradient during exercise. We have studied by exercise test a group of patients successfully operated on for isolated coarctation of the aorta to evaluate the behaviour of the systolic blood pressure and the upper/lower limb pressure gradient during exercise. The aim was to recognize the patients who, inspite of very good operative result, remain at cardiovascular risk in the long-term follow-up. PMID- 10687158 TI - [Celiac disease and lymphoma]. AB - Celiac disease (CD) has been acknowledge as being responsible for numerous secondary pathologies, in particular autoimmune and neoplastic diseases. Whether CD is more prevalent in patients with non-Hodgkin's lymphoma (NHL) than in the normal population is not known. Accordingly, we carried out a study of 86 patients hospitalized in the Section of Oncology, Haematology and Internal Medicine of the Department of Medical, Oncological and Radiological Sciences of the University of Modena and Reggio Emilia and who, between 1988 and 1995 had been diagnosed as affected by NHL. On diagnosis, and before the beginning of antitumour therapy, all the patients were tested for antigliadin (AGA IgA and IgG) and antiendomysium (EMA) antibodies together with total class IgA antibody levels. Our findings showed that none of the 86 patients had an IgA deficit, while one tested positive for AGA IgA (43.9% v.n. < 7.5). The same patient also tested positive for EMA. The extremely high sensitivity and specificity of the AGA IgA and EMA led us to conclude that the patient was affected by CD, although his early death precluded confirmation by biopsy. The presence of one celiac patient among 86 NHL patients examined at the onset of the disease would suggest that CD is not infrequent in NHL. The numbers involved in our study are insufficient for statistical purposes, and we are therefore awaiting the results of a SIGEP multi-centre study into the connection between CD and lymphomas. PMID- 10687159 TI - Scintigraphic evaluation of gastroesophageal reflux in newborns. AB - OBJECTIVES: This study was carried out to verify whether during neonatal stage gastroesophageal scintigraphy, may be useful not only to diagnose gastroesophageal reflux (GER), but also to detect pulmonary aspiration, at times responsible for respiratory symptoms frequently associated with GER. PATIENTS AND METHODS: Gastroesophageal scintigraphy was performed on 50 newborns admitted to NICU of Brindisi in the last two years who presented symptoms as cyanosis, apnoea, bradycardia, laringeal stridor, wheezing, not related to respiratory or cardiac problems, but suspected to be clinical suggestive symptoms of "pathological" GER. After administration of 99mTC mixed with the usual meal formula, we obtained a recording period of 60-90 minutes and a later recording at 4 and 24 hours to document radioactive material in lungs. RESULTS: Scintigraphy showed GER in 40/50 cases and despite frequent observation of respiratory symptoms (39/40 cases) in these newborns never radiation in pulmonary fields. CONCLUSIONS: Our results could indicate that scintigraphy, noninvasive and low in radiation, may be considered effective and reliable to diagnose GER in newborns. The absence of radiation in pulmonary fields could indicate that respiratory symptoms frequently associated with GER could depend on involuntary mechanism of vagal type and not on pulmonary aspiration. PMID- 10687160 TI - Study on 480 hospitalized febrile children: evaluation of the septic risk and results of the antibiotic and corticosteroid combined therapy. AB - The febrile child, previously healthy, represents a frequent diagnosis and management problem for pediatricians who work in private offices and those in hospital emergency departments. We are specifically interested in the identification, for the febrile child with septic risk, of severity parameters permitting to assess the likelihood of a serious bacterial infection. In the retrospective study we present, carried out on children admitted for febrile illness, two factors were mainly evaluated: 1. identification of the septic risk on the basis of some laboratory and clinical parameters; 2. effectiveness of an early antibiotic plus corticosteroid treatment in a sub-group of children with septic risk. The parameters we considered have been the degree of temperature upon admission, the clinical appearance and the C-reactive protein (C-RP) values. A significant correlation resulted for levels of temperature over 39 degrees C, toxic-appearing child and very positive C-RP values. We have defined this condition as a "threatening" fever requiring an immediate hospitalization in order to administer appropriate blood tests and cultures, and also, according to our rationale, to start an early antibiotic plus corticosteroid therapy (within 6 12 hours from the disease onset). PMID- 10687161 TI - [Use of mini-laparoscopy in intraoperative diagnosis of contralateral inguinal hernia in children]. AB - Laparoscopy has been considered by some Workers an useful means of diagnosing patent processus vaginalis in children. This technique is effective in evaluating patency of contralateral internal inguinal ring and requires only five minutes of additional operative time to elective inguinal herniorrhaphy. The Authors report their experience with fifty-six children to whom this procedure was offered. Their age ranged from three to ten years--mean 6.5 years-. Congenital unilateral hernia was right-sided in eighteen and left-sided in thirty-eight patients. Overall, the contralateral processus vaginalis was patent in 44.6%, with 27.7% and 52.2% patency on the nonclinical right and left sides, respectively. Low abdominal pressure-insufflation with CO2 to a pressure of 4 mmHg and shortened operative time have permitted to avoid endotracheal intubation. We confirm great utility of laparoscopic examination in determining the need for contralateral inguinal exploration in pediatric patients. PMID- 10687162 TI - [Evaluation of diosmectite in acute diarrhea in children]. AB - The aim of this case-control study, was to estimate clinical effects, safety and parents' assessment of Diosmectite, in acute infantile diarrhoea. Thirty-five children aged 0-5 years, with acute diarrhoea from at least 5 days received alternatively oral rehydration solution (ORS) and Diosmectite (3 g/day for 3 days in children aged under 3 years and 6 g/day for 3 days in children aged over 3 years) (16 cases) or ORS only (19 cases). At discharge, parents' opinion concerning satisfaction for treatment received was evaluated through a questionnaire. Differences in the amount of liquid intake, weigh recovery, resolution of fever, resolution of vomiting and number of loose stools in the first 48 hours of hospitalization were not significant. Diosmectite was well tolerated and no major side effects were found. Parents' satisfaction for therapy resulted higher in the treated group than in the control group, even if differences were not significant. Treatment with ORS and Diosmectite allowed to shorten the mean duration of hospitalization by 0.4 days/children in the treatment group compared to the control group. Further studies with more cases and with higher does of the drug are necessary to confirm our results. PMID- 10687163 TI - [Acute gouty arthritis in adolescents with renal transplants]. AB - Hyperuricemia is a common metabolic abnormality in subjects with renal transplantation: in fact in transplanted adults receiving immunosuppressive and diuretic drugs the frequency of hyperuricemia varied from 30 to 84% according to treatment. Conversely, the gout is an uncommon eventuality, representing less than 10%; predisposing factors are impaired renal function and older age. In the younger patients with renal transplantation hyperuricemia is also frequent, but the gout doesn't considered a possible complication in paediatric age. We reported our observation of 5 patients (3 males and 2 females), 13-18 years old who developed gout 2-84 months after renal transplantation. All the patients were receiving cyclosporine, 4 even with prednisone and azathioprine. Two patients were treated with furosemide because hypertension. The average of uric acid serum levels in the post transplantation follow-up was 7 +/- 2 mg/dl; at the moment of gout attack the uric acid serum levels raised to 12 +/- 1 mg/dl. The arthritis diagnosis were made by clinical, laboratory and instrumental data (Rx and US). In the most severe cases, uricasi therapy resolved clinical picture. The analysis of immunosuppressive and diuretic treatment, renal function and dietary uses induces us to think that the gout episode may be the result of many concomitant factors, in adolescents with renal transplant. PMID- 10687164 TI - [Giant posterolateral congenital extra-trigonal diverticula of the bladder: an uncommon nosologic entity]. AB - The Authors, on the basis of their observation of two cases of giant posterolateral congenital extratrigonal diverticula of the bladder (D.G.P.P.L.E.), identify it as a nosological entity apart in the widest category of bladder's diverticula, which implies a peculiar diagnostic and therapeutic attitude. PMID- 10687165 TI - [Femoral hernia in childhood: a rare entity]. AB - The Authors present 4 cases of femoral hernia treated in their pediatric surgery department during the last 11 years. Childhood femoral hernia is rare and often missed clinically because of difficulty in eliciting their clinical signs. During an operation for inguinal hernia it is essential to think of femoral hernia when a non obliterated processus vaginalis is not found. PMID- 10687166 TI - [Report of case of Prader-Willi syndrome]. AB - We reported a case of Prader-Willi Syndrome. The simple chromosomic mapping is negative. The obesity and polifagye is not present (2 years old). The ipotony is improving, this case belongs to a possible light case. PMID- 10687167 TI - [Effect of the NMDA receptor antagonist on parameters of delayed visual differentiation and rearrangement of the impulse activity of neurons in visual and prefrontal cortices in monkeys]. AB - With the aid of a multichannel leads the unit activity was recorded in the 17th and 8th cortical fields along with registering behavioural data in Rhesus monkeys. Multi-factor variance analysis revealed that the 2-amino-5-phospho valeric acid (APV) effect involved a significant worsening of the monkeys' behavioural characteristics: duration of the short-term memory shortened (2-4 fold), motor reactions' time increased, and the changes of cognitive characteristics were always followed by significant rearrangements of the unit activity in the above areas. The data obtained suggest that these cognitive dysfunctions are due to a desynchronisation of unit activity in different areas of the cortex including the neuronal assemblies maintaining the short-term memory mechanisms associated with the glutamatergic structures. PMID- 10687168 TI - [Effect of the unilateral activation of brain hemispheres on visceral pain sensitivity in BALB/c mice]. AB - A hemispheric asymmetry of the visceral pain sensitivity control was revealed in the BALB/c mice: animals with the left hemisphere inactivation did not differ from sham-operated control mice in respect to the pain response parameters. A right hemisphere inactivation reduced or suppressed the pain response. This suggests that the right hemisphere domineers in the visceral pain control. PMID- 10687169 TI - [Effect of the preparation baliz-2 on the integrative brain activity and biogenic amine metabolism in rats under stress]. AB - Effects of balis-2 on exploratory activity in the open field and elevated plus maze, attention to sensory stimuli of different modalities, elaboration and retention of conditioned reflexes with food reinforcement, were studied in rats under stress. Prolonged treatment of rats with balis-2 seems to normalise integrative activity and metabolism of serotonine and dopamine in the animal brain. PMID- 10687170 TI - [Application of the simplified method of optic recording for mapping focuses of the neuronal activity in the somatosensory cortex of the white rats]. AB - In rats, a differential signal was used for evoking an optical image in the barrel-field zone which represented a difference between cortical images during the control and the stimulation periods. After a subtraction of averaged sequences of frames, an image of spots reflecting a probable location of activated groups of neurones was obtained. Natural low frequency stimulation of vibrissae is supposed to be the most effective. The method of intrinsic optical activity imaging can be applied for preliminary mapping of cortical zones. PMID- 10687171 TI - [Vascular reactivity and thrombus formation during postischemic reperfusion]. AB - Reactivity, thrombogeneity, and thromboresistance of the rat mesentery microvessels were studied in postischemic reperfusion of the intestine, the brain, an extremity. Irrespective of ischemia localisation, an augmentation of the microvessels reactivity and reduction of their thromboresistance, were found. The microvessels thrombogeneity was depended on the ischemia localisation: an augmentation of the thrombogenity occurred in arterioles whereas it was reduced in venules following the brain and intestine reperfusion. A possible mechanism of the phenomenon may involve a deficiency of the nitric oxide synthesis. PMID- 10687172 TI - [Dependence of the pump function of the isolated rat heart on the functional activity of sigma receptors during reperfusion]. AB - Pre-treatment of the sigma-receptor with the sigma-receptor agonist (+)-SKF 10.047 improved the reperfusion recovery of cardiac pump function. The sigma receptor activation, among other effects, prevented the reperfusion contracture, increased pressure in the left ventricle, and improved survival of cardiomyocytes after ischemia/reperfusion. Pre-treatment with the sigma-receptor antagonist DuP 734 augmented the reperfusion systolic dysfunction of the myocardium and prevented postischemic contractures and cardiac cell lesions. Activation of the cardiac sigma-receptor seems to prompt an augmentation of tolerance to the reperfusion damage. PMID- 10687173 TI - [Dependence of the protective effect of the elevated sodium level on the type of oxidative substrate in the isolated heart during "calcium paradox"]. AB - Isolated guinea pig heart were perfused with the Tyrode solution followed in 15 min. by a 10-min. Ca(2+)-free solution with subsequent return to the normal Ca(2+)-containing Tyrode solution. Sarcolemma damage was measured by myoglobin release. The perfusion resulted in damage of the myocardium cells. The data obtained show that elevation of the extracellular pressure during reperfusion with the Ca(2+)-containing medium is more important than the absolute value of the osmotic pressure. PMID- 10687174 TI - [Effect of the sciatic nerve blockade on the function of the contralateral motor center in the rat gastrocnemius muscle]. AB - When the rat spinal motor centre was activated by fixing the animal in a supine posture, the motor units of intact gastrocnemius muscle fired more frequently in the high-frequency range. Under conditions of the ischiatic nerve blockade, a shift to an increase in the background firing frequency of the motor units related to intact gastrocnemius muscle seems to be related to increased motoneurone excitability occurring because of the contralateral denervation. PMID- 10687175 TI - [Effect of modification of the structure of interleukin-1beta on its physiological activity]. AB - Modification of the structure of recombinant human IL-1 beta: deletion of the amino acids serine, asparagine, and asparagic acid in position 52-54 in mutant delta SND, led to major changes in its functional activity. Significant reduction of the main IL-1 beta activities: concomitant, pyrogenic and corticotropic ones in the mutant delta SND, suggests the latter to be a partial agonist of IL-1 beta. Nevertheless the reduction in certain parameters of the humoral immune response in mice following administration of the delta SND preparation suggests that, when manifesting its immunomodulatory properties, the delta SND acts as a partial antagonist of the IL-1 beta. PMID- 10687176 TI - [Changes in secretion of sex steroid hormones during stress in rats with various brain excitability]. AB - A stress procedure decreased the plasma progesterone faster in highly excitable rats than in the less excitable those. The decrease in gonadal and adrenal progesterone was still obvious in 24 hours in both groups of rats but more obvious in the former group in respect to the progesterone level. The data obtained may help to understand individual ability to respond to a stress in a different way depending on functional condition of the hypothalamo-pituitary adrenal axis. PMID- 10687177 TI - [Characteristics of responses of the sympatho-adrenal system to various types of cooling]. AB - Fast cooling involving the dynamic activity of the skin cold receptors seems to establish a condition for changes in catecholamine concentration at a lesser decrease of body temperature as compared with slow cooling. PMID- 10687178 TI - [Effect of tobacco smoke on the level of estrogens and DNA in the rat uterus]. AB - Tobacco smoke induced no changes in the rat uterus weight or in oestrus cycle but decreased estradiol (E2) concentration in the uterus tissue and increased and later decreased the proliferation index and percentage of the cells in the S phase. The data obtained suggest a phasic character of changes in the reproductive system under the effect of tobacco smoke and corroborate the concept of the role of smoking in the shifting the type of hormonal carcinogenesis from promotional to genotoxic one. PMID- 10687179 TI - [Dynamics of oxyhemoglobin, lactate, and pyruvate in plasma in connection with the woman menstrual cycle]. AB - Levels of lactate, piruvate, oxy- and carbohaemoglobin were shown to change regularly during women's menstrual cycle reflecting changes in metabolism in respect to forming and destroying of the ovum. PMID- 10687180 TI - [Effect of dimedrol on the regulation by histamine of the pacemaker activity of the cat ureter]. AB - A certain concentration of Dimedrol (0.5 x 10(-6) mol/l) turned out to be capable of neutralising the histamine regulation of the slow-wave activity. The histamine H1-receptors were shown to modulate the cat general pace-maker's activity as well as the Na+ mechanism of the slow wave genesis. PMID- 10687181 TI - [Neurophysiological correlates of delayed visual differentiation in monkeys: effect of location of the NMDA-receptor intracortical blockade]. AB - A multi-factor variance analysis revealed that the 2-amino-5-phosphonovalerian acid (APV) effect was manifested in monkeys by a decrease in the number of correct responses entailing a two-fold shortening of the short-term information storage, as well as by an augmentation of the motor responses' time. The correct responses' probability depended on the APV diffusion localisation in the cortex whereas the motor responses' time did not depend on it. The APV effect was accompanied by a desynchronisation of the unit activity in fields 46 and 17. Neurophysiological correlates of cognitive functions, are discussed. PMID- 10687182 TI - [Pharmacologic analysis of the subunit composition of AMPA-receptors in the hippocampal neurons]. AB - Subunit composition and abundance of flip version of different AMPA receptor subunits were studied in neurons acutely isolated from hippocampal area CA1 and dentate gyrus. Whole cell recordings were made to record kainate unduced currents. Presence of GluR2 in the receptor complex led to significant decrease of selective channel blocker IEM-1460 potency. Flip versions of AMPA receptor subunits were discriminated on the basis of their sensitivity to cyclothiazide. Principal cell AMPA receptors in both areas were characterized by low sensitivity to IEM-1460 while AMPA receptors of nonprinciple cells exhibited high or intermediate sensitivity to IEM-1460. We observed significantly larger potentiating effect of cyclothiazide on principal cells. Our data indicate that there is a correlation between low sensitivity to IEM-1460 and high sensitivity to cyclothiazide among AMPA receptors of different cells. Principal cells in both regions possess more GluR2 subunits in their AMPA receptor complexes and more abundant flip versions of their subunits in comparison with nonprincipal cells. This correlation is obviously related to functional pecularities of different neurons. PMID- 10687183 TI - [Current concepts of involvement of glutamate in ischemic damage of the brain tissue]. AB - Glutamate neurotoxicity cannot be considered the only reason for neuronal damage. Other neurotransmitters and molecular messengers like nitric oxide, may be involved in the toxic effect of glutamate. Different conditions in focal and global ischemia, stage of the ischemic damage evolution, the degree of ischemia, and activation of glutamate receptors, should also be taken into consideration when assessing a neurological outcome. PMID- 10687184 TI - [Blood pressure increase in foster mothers in ISIAH and Wistar rats: effect of reciprocal cross-fostering]. AB - Maternal arterial blood pressure (ABP) of hypertensive (ISIAH) and normotensive (Wistar) rats rearing their natural litter or litter of opposite strain, was assessed when the pups were 1.5-, 3- and 4-month old and compared with the ABP of these rats prior to their mating. The ABP was increased in both breeds of rat strains for either cross-fostered or infostered pups. No ABP changes were observed in control rats. PMID- 10687185 TI - [Regional difference in endothelium-dependent vascular responses upon the flow rate elevation]. AB - In anaesthetised rabbits, femoral artery manifested a maximal reserve capacity in its vasodilatory response to the flow speed acceleration. In order of the capacity diminishing, this artery is followed by renal artery, abdominal aorta, celiacus tube, common carotid artery. The endothelium-dependent flow-induced response is important for restricting the flow's linear speed. PMID- 10687186 TI - [Changes in hypoxic resistance and conditional avoidance reflex during hypobaric adaptation on the "flatland" and in mountains]. AB - White rats--males with mass 200-250 g were exposed ipobaroadaptation (rise on 12,000 m with an exposition 5 minutes) with periodic testing them on gipocsition stability. After shaping in the result of gipobaroadaptation stressostability, that it was judged in 2-3 times enlarging of reserve time, rats produced conditional reflexes at active avoid (CRAA) as on plain area (760 m) so in high mountainous (2500 m). It is observed that gipobaroadaptation increases gipocsition stability and accelerates the CRAA shaping on plain area and in mountains. The maximum effect of gipocsition stahility increasing, CRAA consolidation and perfecting of efferention synthesis at high-mountainous gipobaroadaptation rats is more significant, thah at "flat." gipobaroadaptation has important protective significance for function of a brain, decreasing its sensibility to lock of oxygen and fixing maximum adaptive responses. PMID- 10687187 TI - [Morphofunctional changes in the placenta and in the hypophyseal-adrenal system during low intensity infrared radiation in rats]. AB - Morphofunctional conditions of hypophysealadrenal axis and placenta following hypoxis hypoxia and an infrared treatment, were studied in rats. The data obtained involved high levels of the DNA, RNA, and lipidogenesis. Probable mechanisms of structural changes are discussed. PMID- 10687188 TI - [Immune responses in C57BL/6J mice with the inverted pattern of behavior during social conflict]. AB - Inversion of aggressive behaviour into a submissive one led to immunosuppression in C57BL/6J mice as well as in those mice which did not change their behaviour, whereas inversion of submissive behaviour into aggressive one resulted in immunostimulation. A possibility to influence immune response changing the brain neurochemical pattern by reversing behaviour under conditions of a social conflict, is discussed. PMID- 10687189 TI - [Study of the chemoreception in mollusca]. AB - Recording of the mollusc osphardium nerve activity revealed the osphardium sensitivity to hyperosmotic pressure, sodium chloride, and aminoacidosis. The osphardium was found to detect quality of the water where the animals were kept. Pont snail osphradium preserves ancestors' multisensority which unites perception of different chemical and physical signals. Patch-clamp method revealed membrane currents in certain ganglia and receptor cells which are sensitive to Na+ and L aspartate increase in the solution around the osphardium. Inward components of these currents are, probably, of the sodium and/or calcium nature, whereas the outward components--of the potassium one. PMID- 10687190 TI - [Seasonal changes in the function of mechanosensitive receptors in the rat skin]. AB - The effect of circannual rhythms on functional properties of the rat skin tactile receptors and nocireceptors, was studied. The effect proved to be quite significant for the cutaneous receptors threshold stimuli and for the parameters of their responses: the latency and AP duration. These changes seem to be determined by hormonal mechanisms. PMID- 10687191 TI - [Ca(2+) level in the animal blood and their cold resistance]. AB - Administration of small doses of the EDTA decreased by 15-20% the Ca2+ contentn in the blood plasma of rabbits and rats. The decrease coincided with an abrupt stimulation of the thermoregulation system of cooled animals. Restoration of the Ca2+ content in circulating blood coincided in time with repeated suppression of the system's functions. The findings corroborate the theory of a key role of the Ca2+ in sensitivity of the homoiothermal organism to cold and substantiates the method of restoring physiological functions in deep hypothermia without rewarming the body. PMID- 10687192 TI - [Myoelectrical activity in the stomach during ulcerogenic exposure in rabbits]. AB - Stomach myoelectrical activity changed the slow wave frequency and spike discharges under the effect of experimental injury in rabbits. High-amplitude fluctuations named the "injury potentials" appeared. Bensohexonium prevented the changes as well as ulceration. Metacin combined with proserin accelerated healing of the ulcer and the rate of stomach myoelectrical activity. PMID- 10687193 TI - [Effect of protein deficiency in the female rat diet during pregnancy and lactation on the activity of the membrane and soluble forms of digestive enzymes in the offspring small intestine]. AB - Protein deficiency in female rats diet during pregnancy and lactation resulted in deceleration of induction of sucrase both forms in the jejunum and ileum; in acceleration of induction of the maltase membrane from in the jejunum; and in suppression of the lactase membrane form in the ileum; in earlier forming of the adult-type distribution of activity of the membrane form of intestinal alkaline phosphatase and in a decrease in activity of the enzyme soluble form. The findings are corroborated by a suppression of activities of the membrane and soluble forms of the small intestine digestive enzymes in 30-day old rat pups fed with a control (adequate) ration starting 21 days after the birth. PMID- 10687194 TI - [Activity of amino- and dipeptidases in the epithelial and subepithelial layers of the small intestine in rats of various age subjected immobilization stress]. AB - Peptidase systems of all the layers of the jejunum and ileum proved to be most sensitive to stress in young rats, whereas in mature and old rats the peptidases' activity was enhanced, particularly in subepithelial layers of the ileum and jejunum. This may be regarded as an adaptive-compensatory response to probable existence of unsplit low-molecular peptides in the subepithelial space due to an augmentation of proteins catabolism and their inflow via stress-damaged membranes of enterocytes. PMID- 10687195 TI - [Effect of strophanthin on the absorption of sodium and short chain fatty acids from the sheep rumen]. AB - In vivo, intraruminal strophantin administration suppressed the sodium absorption by up to 50% in the sheep reticulo-rumen. Absorption of the shortchain fatty acids was not affected. The findings suggest absence of a combined transport of sodium and short-chain fatty acids' anions in basic and lateral membranes of the reticulo-rumen apithelium in sheep. PMID- 10687196 TI - [Educational-research-practical association "Outpatient therapy" (results and perspectives)]. PMID- 10687197 TI - [Prospective trial of 40-59-year-old males with angina pectoris and essential hypertension (results of 30-35-year prospective studies)]. AB - AIM: To investigate remote prognosis for patients with coronary heart disease and essential hypertension with consideration of the initial total serum cholesterol (TSC) level. MATERIALS AND METHODS: Throughout 30-35 years 103 male patients aged 40-59 years with angina pectoris and essential hypertension were examined (repeated evaluation of protein and lipid spectrum, normal resting and two-step exercise ECG). At the entry, the patients had no history of myocardial infarction (MI), cerebral thrombosis, heart failure, discirculatory encephalopathy. Causes of death were verified by the data of autopsy (70.3%) and registry office's records. Adequate data were obtained on 93 of 103 patients (90.3%). RESULTS: By the end of the trial 89 of 93 patients died. 78 (87.6%) of them died of MI or stroke. 44 deceased from the subgroup with low TSC level lived, on the average, 65.9 +/- 1.3 years, while 45 deceased from the subgroup with high TSC lived 67.9 +/- 1.4 years, i.e. 2 years longer. The worst life prognosis was for patients who developed cancer. They lived 60.5 +/- 5.1 years, i.e. 7.5 years less than the rest of the deceased. In the subgroup with low TSC level there were 5 times more deceased due to cancer than in the subgroup with high TSC level. CONCLUSION: Patients of middle and old age with coronary heart disease require a strictly individual approach to correction of their lipid metabolism and special alertness for cancer in low TSC level. PMID- 10687198 TI - [Prevalence of cardiovascular diseases in organized groups of employees and results of long-term multifactorial prevention]. AB - AIM: Evaluation of cardiovascular diseases (CVD) prevalence and their dynamics after active prevention program. MATERIALS AND METHODS: 1382 male and 155 female volunteers were included into the group of active prevention in their office. 1024 males and 776 females with natural history were compared as a control group. 5-year follow-up was performed in 543 males and 569 females in comparison with 97 males and 119 females. 10-year follow-up covered 82 males and 191 females from the active prevention group and 178 males and 118 females from the referent group. RESULTS: The risk factor correction has resulted in a significant decrease in both systolic and diastolic BP mean levels, prevalence of ECG abnormalities, smoking habits, overweight and combination of the risk factors, general mortality rate, CHD and stroke mortality, number of days of temporary disability. CONCLUSION: The primary and secondary "in office" prevention for 5 to 10 years diminishes the prevalence of the risk factors. The temporary and primary disability rates were connected with the presence of CVD. CVD, in line with other risk factors, significantly increased cardiovascular and general mortality rates. PMID- 10687199 TI - [20-year monitoring of acute cardiovascular diseases in population of large industrial city in West Siberia (epidemiological study)]. AB - AIM: To reveal trends in incidence rates of acute cardiovascular diseases (ACD) in a large industrial city of the West Siberia. MATERIALS AND METHODS: Studies on WHO programs "Acute Myocardial Infarction Register" and "MONICA" have been performed in three districts of Novosibirsk. The diagnostic categories were detected without difference. The observation covered stable population of 500,000 residents aged 25-64 years. Trends in the myocardial infarction (MI) mortality, morbidity and lethality were analysed for 1977-1996. RESULTS: The above trends were stable except for 1986 when MI mortality, morbidity and lethality decreased and 1988 and 1994 when they went up. The reduction was due to 7-year prevention program while the rise was consequent to discontinuation of the preventive measures. Major risk factors of ischemic heart disease, according to screenings conducted in 1984, 1988 and 1994 remained at about the same level. Social stress closely correlates with a rise in MI morbidity and mortality. The latter in 1994 grew owing to higher rates of MI mortality and morbidity among the oldest men and females of different age groups. CONCLUSION: Urgent intensification of prophylactic measures is needed both at the populational level and the level of high risk strategy. PMID- 10687200 TI - [Clinico-psychological features of patients with angina pectoris and obesity]. AB - AIM: To study clinical and psychological features of patients with coronary heart disease (CHD) and their comparison with obesity. MATERIALS AND METHODS: The presence and intensity of cardialgia, Ketle's index, body mass fat component, tolerance to isometric load were determined in 104 patients with stable angina of effort and 42 healthy males. Questionnaire surveys were performed. 46 obese patients were reexamined in 7-8 months. RESULTS: Mean body fat mass in anginal patients was higher than in the controls. Obese patients were more anxious, psychosocially maladapted, less tolerant to isometric exercise, had more frequent cardialgia. The attitude of the patients to body mass reduction and its dynamics depended on personal traits. CONCLUSION: Obesity was observed in the majority of anginal patients (60.6%) and is closely related to their clinicopsychological characteristics. PMID- 10687201 TI - [Program of medical and psychological support of patients after cardiosurgery]. AB - AIM: To study effectiveness of the program of medical and psychological support of patients after aortocoronary by pass operations (ACBO). MATERIALS AND METHODS: The trial included 108 patients with coronary heart disease (CHD) after ACBO. 72 patients of the test group participated in the program of medical and psychological support. 36 patients who did not receive such support served control. All the patients underwent clinicophysiological and psychological tests aimed at assessment of exercise tolerance and personality types. RESULTS: Many of the patients after ACBO need psychological consulting and support. The course of group therapy gives the patients strong motivation for following the doctor's recommendations and health self-control. Use of this motivation produced improvement in psychosomatic health of the test group, increased the percentage of patients who resumed their previous jobs. CONCLUSION: The program of medical and psychological support in CHD patients after ACBO increases the effectiveness of their rehabilitation and psychosocial activity, quality of life and return to previous job. PMID- 10687202 TI - [Association of lipoprotein(a) and apolipoprotein(a) phenotypes with coronary and carotid atherosclerosis in CHD men]. AB - AIM: To evaluate in a case-control cross-sectional study whether lipoprotein(a) concentration and apo(a) phenotypes are associated with the presence and severity of coronary and carotid atherosclerosis. MATERIALS AND METHODS: We have examined 198 male CHD patients (mean age 53 +/- 8) years) with stenosis more than 50% at least in one main coronary artery or its major branches. Duplex scanning was performed in 168 patients to assess the degree of carotid atherosclerosis. Seventy six apparently healthy men (mean age 39 +/- 9 years) formed the control group. Lp(a) concentration was measured by ELISA, apo(a) phenotyping was performed by immunoblotting. RESULTS: Lp(a) level was significantly higher in cases compared to controls: 37 +/- 31 mg/dl vs. 18 +/- 27 mg/dl, p < 0.05. Patients had low-molecular weight apo(a) phenotypes more frequently than controls: 46% vs. 29%, p = 0.01. Patients aged 45 years and younger had low molecular weight apo(a) phenotypes more frequently than older ones (65% vs. 42%, p < 0.05) and controls (65% vs. 29%, respectively, p = 0.001). High Lp(a) level and low-molecular weight apo(a) phenotypes correlated with presence and number of coronary occlusions. CONCLUSION: There was association between Lp(a) level, low molecular weight apo(a) phenotypes and presence, severity, extension of carotid atherosclerosis. No differences in distribution of other CHD risk factors among all subgroups of patients were found. PMID- 10687203 TI - [Prevalence and intensity of smoking among adult urban population of Tiumen': data of population mail questionnaire survey]. AB - AIM: To estimate prevalence and intensity of smoking among adult Tyumen citizens by mail questionnaire survey. MATERIALS AND METHODS: A representative sample of adult population of Tyumen (central district) was randomly selected using the list of the electors. The questioning covered a total of 3200 citizens (8 groups per 400 age- and sex-matched subjects). The response reached 71.5%. Smoking in the population was studied in three aspects: prevalence, intensity and passive smoking. RESULTS: Overall prevalence of smoking was not great (28.8%), but smoking was frequent in women, especially young, was intensive in males of all the age groups. The number of smokers who had quit smoking was small. Passive smoking at jobs was prevalent. CONCLUSION: The data of the survey indicate difficulties of social adaptation for the studied population who fall under risk to develop cardiovascular diseases. PMID- 10687204 TI - [Features of dopaminergic system function in menopausal females with arterial hypertension]. AB - AIM: Assessment of dopaminergic activity by changes in urine excretion of dopamine, arterial hypertension, blood prolactine, 24-h diuresis and natriuresis in response to a single (2.5 mg) and 7-day (daily dose 5 mg) administration of parlodel in patients with essential hypertension (EH) in menopausal and reproductive age having different serum levels of estrogens. MATERIALS AND METHODS: The indices of 52 hypertensive menopausal women and of 18 women at reproductive age of whom 8 had hypothalamic syndrome were followed up. RESULTS: A single dose parlodel (bromocriptine) in the dose 2.5 mg 2 hours after its administration caused a significant fall in arterial pressure, rise in 24-h diuresis in hypertensive menopausal women (p < 0.01). Parlodel given in a daily dose 5 mg for 7 days in menopausal hypertensive women lowered a significant fall in arterial pressure, blood concentrations of aldosteron, prolactin, aroused dopamine and urinary sodium excretion (p < 0.01). Similar were the changes in hypertensive women at reproductive age with hypothalamic syndrome (n = 8). In such women free of hypothalamic impairment the changes did not occur (n = 10). A significant positive correlation was found between serum levels of prolactine and estradiol in all the hypertensive women in menopause. CONCLUSION: Deficiency of dopaminergic activity in menopause is induced by hypoestrogenemia correlating with serum levels of prolactin. This deficiency was identified in hypertensive menopausal women by the results of acute and prolonged tests with dopamine mimetic--parlodel. PMID- 10687205 TI - [Effect of enalapril maleate on vascular endothelial function and platelet endothelial interactions in patients with essential hypertension]. AB - AIM: Evaluation of endothelial function and platelet-endothelial interactions in patients with essential hypertension and dynamics of these changes in the course of treatment with enalapril maleate. MATERIALS AND METHODS: The study included 37 patients with essential hypertension and 22 normotensive volunteers. 17 of hypertensive patients received enalapril maleate (enap, KRKA) 5-20 mg/day during the period of 1.5 months. The complex of investigations included: measurement of total plasma cholesteroi, 12-lead ECG, echocardiography, high-resolution ultrasound investigation of brachio-cephalic arteries, evaluation of flow mediated dilation, measurement of von Willebrand's factor, spontaneous and induced platelet aggregation. RESULTS: Patients with essential hypertension exhibited higher levels of von Willebrand's factor in plasma and degree of spontaneous and induced platelet aggregation as well as lower responses of vessel wall to hemodynamic stimuli compared to normotensive healthy individuals. There was a strong correlation between endothelial function markers and CAD risk factors, elevation of platelet activity. Treatment with enalapril maleate led to a statistically significant decrease of von Willebrand's factor in plasma and ex vivo platelet aggregation whereas flow-mediated dilatation increased. Values of endothelial function markers and platelet activity approached to those of normotensive subjects and these changes were accompanied by a decrease of ECG signs of left ventricular hypertrophy. CONCLUSION: Patients with essential hypertension were found to have compromised endothelial function. However, the degree of endothelial dysfunction depends not on hemodynamic parameters, but on the cumulative effect of CAD risk factors. Treatment with enalapril maleate may lead to normalisation of endothelial function and decrease of platelet activity. PMID- 10687206 TI - [Metabolic effects of quinapril in hepertensive patients]. AB - AIM: The study of antihypertensive and metabolic effects of quinapril (accupro) in patients with essential hypertension (EH). MATERIALS AND METHODS: Serum levels of cholesterol, its fractions and triglycerides were measured and standard glucose tolerance test was made in 40 EH patients before accupro therapy and 1 and 3 months after continuous accupro treatment (10-20 mg/day). RESULTS: Antihypertensive effect of accupro was accompanied by marked improvement in carbohydrate and lipid metabolism in 28 patients. Normalization of blood pressure in the absence of negative effect of accupro on parameters of carbohydrate and lipid metabolism was recorded in 12 patients free of coexisting metabolic disorders. CONCLUSION: The results of the trial allow to recommend antihypertensive treatment with accupro as effective both in patients with metabolic disorders and free of them. PMID- 10687207 TI - [Some approaches to raising quality of treatment for arterial hypertension (experience of school for arterial hypertension patients)]. AB - AIM: To investigate involvement of patients into control of their pressure to improve quality of treatment for arterial hypertension (AH). MATERIALS AND METHODS: 134 hypertensive patients (76 females and 58 males) were interviewed. Criteria of efficacy of AH control were ability and motivation of self measurement of blood pressure and its maintenance at the optimal level. RESULTS: For the past year, 40% of the patients measured pressure regularly (once a day or more frequently), 34%--1-3 times a week, 9%--1-3 times a month, 17%--rarely, mostly, males. Most of the patients controlled their pressure insufficiently. This is explained by "working pressure" (high in many cases) which was registered in most of the examinees. When part of the patients reached that working pressure they discontinued treatment. CONCLUSION: Involvement of hypertensive patients and members of their families into control of hypertension is a feasible and valuable method of improving quality of hypertension treatment. PMID- 10687208 TI - [Clinicofunctional features of arterial hypertension in chronic broncho obstructive syndrome]. AB - AIM: To describe clinicofunctional features of essential and pulmogenic hypertension in chronic bronchoobstructive syndrome, 24-h profile of arterial pressure (AP), intracardiac hemodynamics and to propose differential diagnostic criteria for these hypertension forms. MATERIALS AND METHODS: 24-h monitoring of arterial pressure (MAP), cardiohemodynamics, external respiration function (ERF) and blood gases examinations were made in 100 hypertensive subjects with chronic obstructive bronchitis and bronchial asthma. RESULTS: Significant differences were found between the groups of essential and pulmogenic hypertension by major values of MAP, echo-CG and ERF. Early disturbances in diastolic function of both the ventricles in essential and pulmonary hypertension in chronic bronchial obstruction arise long before clinical, x-ray and ECG signs of chronic pulmonary heart. CONCLUSION: The findings enable formulation of criteria of differential diagnosis of essential and pulmogenic hypertension and identify the latter as an independent nosological entity. PMID- 10687209 TI - [Prehospital treatment of myocardial infarction]. PMID- 10687210 TI - [Arthrotec treatment of rheumatoid arthritis]. AB - AIM: To try arthrotek, a combined drug, in patients with rheumatoid arthritis (RA). MATERIALS AND METHODS: 10 RA women aged 30 to 60 years with endoscopically verified minimal affection of gastroduodenal mucosa (not more than 25 hemorrhages and/or 5 erosions) received arthrotek (1 tablet 3 times a day for 3 months). The efficacy was judged by changes in the joints and gastroduodenal lesions. RESULTS: The 3-month treatment produced positive changes in the main auricular and gastroduodenal symptoms. Side effects (head ache, diarrhea, sleepiness) disappeared after reducing the day dose to 2 tablets. CONCLUSION: Arthrotek proved highly effective against RA. It is also a good gastroprotector promoting healing of gastroduodenal erosions. PMID- 10687211 TI - [Acute leukemia in adults: morbidity in Donetsk region of Ukraine before and after Chernobyl' accident]. AB - AIM: To study whether Chernobyl accident has implications for acute leukemia (AL) incidence rate in Donetsk region. MATERIALS AND METHODS: Records of the Donetsk regional hematological center for new verified cases of AL have been analysed for 1977-1987 and 1989-1994. RESULTS: Four years after the accident AL morbidity was higher than before the accident. After 1991 AL morbidity returned to the preaccident level. CONCLUSION: A rise in AL incidence is attributed to radionuclide contamination after the Chernobyl accident of a region with initially unfriendly environment. PMID- 10687212 TI - [Peculiarities of cell reception in bronchial asthma patients with bronchial hyperreactivity to hyperosmolar provocation]. AB - AIM: To investigate characteristics of the cell receptor systems in bronchial asthma (BA) patients with bronchial hyperreactivity (BHR) induced by hyperosmolar provocation. MATERIALS AND METHODS: 15 patients with BA, in most cases atopic, in remission were studied. The bronchoprovocative test (BPT) with ultrasonically nebulized 3.6% hypertonic NaCl solution (UNHS) FEV1-control was performed. Cell receptors reactivity was analysed on the model of erythrocyte resistance (ER) to hyperosmolar provocation (HP) under the modulation by adrenergic and histaminergic agents. RESULTS: It was revealed that ER was significantly higher (p < 0.05) in BA patients with BHR to UNHS under histaminergic modulation, but ER did not differ in BA patients with or without BHR to UNHS under the adrenergic modulation. The correlation was found between DFEV1 on BPT and magnitude of ER to HP under the blockade of H1-histaminergic receptors (r = 0.72; p < 0.02). CONCLUSION: The development of BHR to hyperosmolar stimulus may be due primarily to disturbance of histaminergic receptor system. PMID- 10687213 TI - [Experience of medical institute therapeutic chair and health service administration activity]. PMID- 10687214 TI - [General practitioners in Orenburg: pilot experience]. PMID- 10687215 TI - [Irritable bowel syndrome (lecture)]. PMID- 10687216 TI - Highlights & trends. The year in review--Part I. PMID- 10687217 TI - [The unification of the postings for medical interns and residents]. PMID- 10687218 TI - [The assessment of quality perceived by the users of the basic health areas of Cuenca]. AB - OBJECTIVES: To find the quality, as perceived by the users, of the health districts of Motilla del Palancar and Cardenete (Cuenca), and define the differences, if they exist, between outlying health clinics and central ones. DESIGN: A descriptive, cross-sectional observation study. SETTING: Health districts of Motilla del Palancar and Cardenete, both in the Cuenca Health Region. PARTICIPANTS: A random sample of 442 persons was selected from the 9542 patients with a health card in the health districts under study. MEASUREMENTS AND MAIN RESULTS: A questionnaire based on the Hulka studies, which had already been used by the Ministry of Health and Consumption in our field and was of proven validity and reliability, was administered to each patient. The main researcher did 442 interviews. The overall assessment by the patients, on a scale from 1 to 7, was 6.07. The scores by patients of the central clinics of the health districts were, in general, lower than those of patients from the rural nuclei. CONCLUSIONS: In general, patients gave high scores in their assessment of perceived quality. Patients in rural nuclei perceived greater health care quality than those from the main clinics. PMID- 10687219 TI - [The perceptions of the adult Spanish population of the factors determinative of health]. AB - OBJECTIVE: To assess which are the most important perceived influential factors on health among the Spanish adult population. This may provide sufficient information to promote primary health care campaigns directed specifically to those that acknowledge that lifestyles are important in spite of not living healthy lifestyles themselves. DESIGN: Observational cross-sectional study. SETTING: Spain (Canary Islands not included). PARTICIPANTS: A representative sample of 1000 Spanish participants over 15 years old selected by a multistage procedure. INTERVENTION: Survey to evaluate the population choosing one of nine known health-related factors (smoking, food, alcohol intake, stress, physical activity, environment, body weight, support from family, genetics) as being one of the two most important factors influencing health through a validated questionnaire. MEASUREMENTS AND RESULTS: The chi-square test for linear trend was used to assess the influence of factors coded on an ordinal scale. The Pearson chi-square test was applied for categorical factors. Smoking was considered the most influential health-related factor by most participants (47.8%). Males, those from lower socioeconomic and educational levels; people living in the south of Spain, and rural regions; those married and individuals who had 3 or more children below 15 years, perceived smoking more often as one of the most important determinants of health. CONCLUSIONS: These data suggest that a large percentage of the Spanish adult population recognizes that life-styles are important determinants of health. PMID- 10687220 TI - [The program for the control of temporary incapacity at the Instituto Social de la Marina of the province of Cadiz]. AB - OBJECTIVE: To evaluate the results of a programme of intervention into the monitoring of short-term unfitness for work of members of the Special Sea Regime at Cadiz during 1997. DESIGN: Quasi-experimental intervention study with a pre- and post-intervention group, formed by the doctors from the Instituto Social de la Marina (ISM) at Cadiz. SETTING: Primary care, 1997. PARTICIPANTS: 21 doctors carrying out their health-care duties in the ISM clinics. INTERVENTION: Activities aimed at the doctors, seeking to highlight the need to protect workers suffering incapacitating pathologies. MEASUREMENTS AND MAIN RESULTS: In January the indices of those off work were similar (2.38 in January 1996, and 2.39 in January 1997). Then the differences widened, with September 1997 (2.42) marking the biggest difference with the same month in 1996 (1.44). However, the mean length of the periods of time off was greater in 1997 than in 1996. The number of sick certificates processed in 1997 (1233) was less than in 1996 (1326). But the opposite occurred with notes for return to work (1209 in 1996 and 1311 in 1997). The mean duration of the period of time signed off increased after the intervention. CONCLUSIONS: A quantitative change in the indicators of management of short-term unfitness for work occurred. PMID- 10687221 TI - [The introduction of generic pharmaceutical products into Galicia]. AB - OBJECTIVE: To know the evolution of the introduction of generic drugs (GDs) in Galicia. Secondarily, to evaluate its potential impact on pharmaceutical expenditure. DESIGN: Descriptive study of GDs utilization. Cost-minimization analysis. SETTING: Galician autonomous region, year 1998. MEASUREMENTS AND RESULTS: Using data from the prescription billing registry of Social Security we have selected the active ingredients corresponding to GDs with prescriptions in Galicia in 1997. We have analyzed the data for their oral single substance preparations by quarters. Consumption in DHDs of allopurinol, atenolol, captopril, naproxen and ranitidine remained stable during 1998. The market share for their GDs in quantitative terms relative to both total consumption of the active ingredients and to their pharmaceutical equivalents, showed an overall growing trend. The maximum observed value was seen for ranitidine at last quarter. Total expenditure (in final customer prices) during 1998 on the selected active substances was higher than 1864 million pesetas. Potential savings afforded by substitution for the lowest price GD prescribed in Galicia would reach 427 million pesetas. CONCLUSIONS: GDs market penetration in Galicia during 1998 was limited but increasing. Its utilization may afford estimated savings of 21-28% of the cost for the selected drugs. However, the expenditure on the above drugs was just 2.7% of total pharmaceutical expenditure. PMID- 10687222 TI - [The social and health variables associated with the self-perception of the health status of the adult population of Gran Canaria (Canary Islands)]. AB - AIM: To find out how the adult population in one municipality of the Canary Islands perceives its state of health, and how this is related to social and health variables. DESIGN: Descriptive cross-sectional study. PLACE: Telde (Gran Canaria). SUBJECTS: The municipality has a population census of 79,000 inhabitants. The sample was multistepped and selected randomly. It included 2626 people with a population variance of p = q = 50%, standard error +/- 4% with a confidence interval of 95.5%. Participation rate was of 82.5%. MEASUREMENT AND RESULTS: Data on aspects of socio-demography, morbidity, medication, use of public health facilities and lifestyles were collected via personal questionnaire. The relationship between different variables and people's perception of their health was calculated by odds ratio using logistic regression. Almost 50% of the sample perceived their state of health to be excellent or good. Subjects over the age of 45 (OR, 2.89), or who were unemployed, perceived themselves to be in a worse state of health (OR, 1.34). Contrastingly, those who had received higher education (OR, 0.39), or who, in the previous 15 days, had not been confined to bed (OR, 0.22), had not been on sick leave (OR, 0.23) or had not visited the doctor (OR, 0.38) had a better perception of their health. CONCLUSION: People's perception of their health is related to their quality of life and self-esteem and continues to be a useful indicator in the evaluation of their general state of health, reflecting aspects of both social level and health. PMID- 10687223 TI - [How we use opioid drugs on patients with neoplasms]. AB - OBJECTIVE: To find the pattern of use of opiate drugs for treating pain in terminal cancer patients. DESIGN: Retrospective descriptive study. SETTING: Gava 2 Health Centre, located in Barcelona's industrial belt. MATERIAL AND METHOD: Systematic review of the clinical records for the deaths recorded between May 1993 and March 1998. The following variables were recorded: age, sex, cause and place of death, professional attending patient during terminal phase, use of opiates (type, how they were taken, dosage and length of treatment) and prescribing doctor. RESULTS: Of the 429 deaths reviewed, 100 (23%) were caused by neoplasm (68% males), with an average age of 69 +/- 3 years. More than half the patients (55%) died at home. In the terminal phase they were mainly attended (69%) by their Primary Care team. 52% of the patients were given opiates, with morphine being the most common (71.1%), followed by codeine (40.3%) and tramadol (17.3%). The general practitioner was the prescribing agent in 69.2% of the cases. 76% of the patients who took codeine did so at infra-therapeutic doses (< 120 mg per day). Similarly, insufficient doses of morphine (< 60 mg per day) were given to half the patients who received it. 21.62% only took it during a period of 5 days or less before death. CONCLUSIONS: Primary care teams are taking on steadily greater protagonism in caring for terminal cancer patients. Although a large number of these patients are treated with opiates, these are given at often insufficient doses and for too short time periods. PMID- 10687224 TI - [Electronic journals in primary care]. PMID- 10687225 TI - [How many more diabetics do I have?]. PMID- 10687226 TI - [The group education of diabetics]. PMID- 10687227 TI - [Antidepressants and sexual dysfunction]. PMID- 10687228 TI - [Clinical changes in patients undergoing group diabetes education]. PMID- 10687229 TI - A clinical review of drug-induced gingival overgrowths. AB - There is an increasing number of medications associated with gingival overgrowth. These medications are used to treat a number of common conditions in the Australian population and as such dentists can expect to manage a number of patients with medication-related gingival overgrowth. This review highlights the clinical features and management of the common overgrowths associated with anticonvulsants, immunosuppressants and the calcium channel blockers. PMID- 10687230 TI - Prophylactic removal of asymptomatic third molars: a review. AB - Mandibular and maxillary third molars are mostly consigned to 'waste bins' in dental practices because they are terminal in developmental timing and positioning in the dental arches, and regarded as functionally non-essential. Thus, many dental practitioners do not attach significance to the presence of third molars when making therapeutic recommendations to patients about preservation of the dentition. The need for taking more a serious account of third molars is reviewed in this paper. PMID- 10687231 TI - Australian dental schools: moving towards the 21st century. AB - In Australia nearly all tertiary education is funded through the Federal Government. With reductions in government spending tertiary education has had to accommodate its share of the cuts. Under such a climate dental schools in Australia face serious financial difficulties, in addition to many other diverse threats, as they head towards the 21st century. Most seriously, and almost uniformly felt, is the diminution of Federal Government funding to a point where the operation of some dental schools remains viable only by way of supplementary funding (direct or in-kind) from State Governments. In this report the authors have developed one possible model of academic, clinical and financial structures of a dental school, based on sound educational and economic grounds, that can overcome some of the short-comings of the paradigm that exists in some schools in Australia. The two key factors underlying the principles of this model for a new style of dental school are flexibility and professional responsibility. Based on the existing academic and economic realities it would be much more appropriate to outsource a significant proportion of the educational and clinical component of a dental school. Highly trained individuals from the dental profession would be invited to provide training in their area of expertise. The role of the dental school would evolve to be like a facilitation centre, organizing the various courses. It would mean that the 'core' curriculum would be the responsibility of the school's academic staff and the outsourced professional members would contribute within the bounds of the basic framework. On the basis of this model a dental school of approximately 225 full-time undergraduate students (50 per year, less some student loss) in a five year programme is planned, and annual staffing costs are estimated at $1.4 million. PMID- 10687232 TI - Hyperbaric oxygen in the prevention of osteoradionecrosis of the jaws. AB - Patients who have had their jaws irradiated as part of management of head and neck malignancy are at risk of osteoradionecrosis (ORN) following tooth extraction. Thirty-seven patients with a history of irradiation to the jaws were managed during a four year period. Twenty-nine patients received hyperbaric oxygen therapy (HBO) consisting of 20 treatments before surgery and ten treatments after. Only one (4 per cent) developed ORN. Seven patients who did not have HBO and one who did (15 per cent) developed ORN. The need for prophylactic treatment with HBO is discussed. It is recommended that prophylactic HBO is used prior to surgery for irradiated facial bones. PMID- 10687233 TI - Cytotoxicity of polymerized commercial cyanoacrylate adhesive on cultured human oral fibroblasts. AB - Cyanoacrylate (CA) has been used as both a commercial and tissue adhesive. Dentists may have had the experience of patients repairing their own acrylic based dentures using a cyanoacrylate (CA) adhesive known as 'super glue'. This study evaluated the cytotoxicity of commercial CA adhesives when fully polymerized, as well as the toxicity of substances released from polymerized commercial CA adhesives after incubation of these materials for various periods of time. Toxicity was tested on cultured oral fibroblasts. Dead cells found around the various CA-coated filter papers constituted inhibitory zones which varied from 200-1000 microns and which persisted for two weeks. Control oral fibroblasts grew to approach the wax-coated filter paper. Cell viability testing using MTT and crystal violet staining methods supported the conclusion that polymerized CA-coated filter paper released substances that are toxic to cells, while wax-coated filter paper gave the same result as the control. The crystal violet staining method was also used to investigate the cytotoxicity of various CA materials after incubation for one, three, seven and 14 days and showed that CA continued to release cytotoxic substances at about the same level for at least two weeks. It can be concluded that, if CA adhesive is used for repair of broken dentures, it will release substances which are toxic to human oral fibroblast cells. This release of substances may persist for at least two weeks. PMID- 10687234 TI - Public knowledge of the prevention of dental decay and gum diseases. AB - In 1992, a mail survey was conducted among South Australians aged 10 years and older to assess the knowledge of prevention of dental caries and gum diseases and to explore its variation by sociodemographic factors. The survey provided 838 completed questionnaires. Questions were asked on perceptions of importance of a number of preventive measures, the main purpose of water fluoridation and sources of information. Respondents rated four myths for preventing dental caries as the most important: 97 per cent rated regular tooth brushing; 87 per cent rated regular dental visits; 85 per cent rated calcium in the diet; and 78 per cent rated eating fibrous foods as definitely or probably important. Only 56 per cent of respondents rated drinking water with fluoride as definitely or probably important for preventing dental caries, and only half (50.2 per cent) identified the main purpose of water fluoridation as the prevention of decay. Respondents rated regular tooth brushing (96 per cent) and regular dental visits (87 per cent) as important for the prevention of gum diseases. However, the myth of massaging the gums was rated as important by 67 per cent. Higher percentages of females, older adults and those with lower educational attainment rated the myths for preventing caries as important. Younger people were less able to specify the main purpose for fluoridation of water supplies. The persistence of myths and the low rating of the importance of scientifically efficacious measures are major challenges for oral health promotion. PMID- 10687235 TI - Child abuse and dentistry: a study of knowledge and attitudes among dentists in Victoria, Australia. AB - Child abuse is a disturbingly common finding in society today. In view of the high proportion of orofacial injuries seen in victims of child abuse, dentists are in a strategic position to recognize and report suspected cases. The present study of 347 dentists in Victoria, Australia, assessed the level of knowledge and attitudes among dental professionals on the important issue of child abuse. While a high level of interest was shown by the participants towards this issue, a need for further information and training in the recognition and reporting of child abuse was seen in the survey findings. While dentists at present are not legally mandated in all states of Australia to report suspected cases of child abuse, the dental profession is in a key position to play an active role in the identification and reporting of this substantial community problem. PMID- 10687236 TI - The use of glass ionomer in special needs patients. AB - Placement of restorations for patients who are physically or intellectually disabled or mentally ill can pose considerable difficulties for the general practitioner. Access to the oral environment is often limited and patient tolerance and concentration may be reduced to rather brief periods of time. Oral hygiene routines may be less than ideal leading to a high caries rate. Enamel surfaces which do not normally become carious can develop broad but shallow lesions with a poorly defined outline. Selection of the most suitable restorative material will be important, with longevity of the restoration as the prime consideration. Other factors such as access, isolation of the lesion and patient co-operation must also be taken into account. Also, forces acting on restorative materials may be less than usual due to poor occlusion, teeth opposing dentures or being completely unopposed. Restoration by indirect techniques will often not be possible so the choice will be limited to the three plastic restorative materials normally used in restorative dentistry: amalgam, resin composite and glass ionomer. As a result of clinical experience it is suggested that glass ionomer will often be the material of choice. This paper describes five years experience with the resin-modified glass ionomers in an institutional practice which is limited to patients with special needs. Clinical significance Placement of restorations, with a reasonable expectation of longevity, can pose considerable problems for the patient with special needs. Resin-modified glass ionomer is a useful alternative material and has been placed with a high degree of success over a period of five years. PMID- 10687237 TI - A programme for the treatment of severe dental fear. Report of three cases. AB - This paper describes the use of a sequence of recollection and discussion, training in muscle tone control, dental imagery, audio simulation and graded re exposure to dental procedures to eliminate a severe fear of dental treatment in three adult female patients. All three patients had abstained from regular dental visits throughout their lives and all were in need of urgent dental treatment when the therapy began. One of the patients also identified in herself other maladaptive behaviour and reported a history of mood disorder as well. The recollections of these patients reveal that the aetiology of dental fear is more complex than dental questionnaires would indicate and, in particular, they supply further evidence of the capacity for adverse medical events to initiate this fear. Although the programme specified in this paper yielded good results, it nonetheless has a number of shortcomings. It is time consuming, dental damage is likely to be well advanced when it commences and it attracts very few patients. Thus those afflicted with severe dental fear are effectively destined for only emergency dental visits and eventually are likely to be edentulous. For this reason every possible precaution against the initiation of dental fear should be taken. PMID- 10687238 TI - Bilateral congenitally missing maxillary canines. A case report. AB - Exclusive aplasia of maxillary permanent canines is extremely rare. There are only a few cases of this condition reported in the literature. This paper reports a case of bilateral congenitally missing canines in a healthy 11 year old male of Chinese origin. The article discusses problems in diagnosis and presents options in the management of such a case. The management option selected was the most appropriate for the family at the time of presentation. PMID- 10687239 TI - Resin ionomer restorative materials for children: a review. Author's reply. PMID- 10687240 TI - Temporomandibular joint arthroscopy. PMID- 10687241 TI - Hydrophilicity? PMID- 10687242 TI - [The production of swine intended for xenotransplants]. AB - Allotransplantation of organs is one of the most prominent medical achievements of this century. The success of allotransplantation has, however, provoked a problem or organ shortage. In order to overcome these problems, the possibility of using animals as potential donors for humans (xenotransplantation) is considered. Many investigators recently proposed the pig as an alternative source of organs. Pigs are easy to breed, have anatomical and physiological characteristics compatible with humans, and are well studied for several pathogens potentially transmissible to humans. Moreover transgenic pigs can be obtained expressing human proteins in order to resist hyperacute rejection. Caesarean-derived piglets can easily be maintained gnotobiotics or specified pathogen free. The pig being nowadays the best candidate for xenotransplantation, this paper will focus on the potential public health risk linked to the use of this species as a source of organs, and the general rules to follow in order to manage it, (risk management). PMID- 10687243 TI - [Regulation of pulmonary circulation by the vascular endothelium]. AB - The function of the endothelial mediators on lung vessels is progressively understood. Our experimental results, in agreement with those from other laboratories, show that prostacyclin and nitric oxide do not affect basal pulmonary vascular tone, but that they limit the pulmonary hypertension secondary to alveolar hypoxia, or to lung injury. The role of endothelins remains harder to define, partly because their vascular effects are opposed by nitric oxide in intact animals. PMID- 10687244 TI - [Inter-species transmission of the influenza virus]. AB - Influenza is an infection of human beings and several animal species. It is caused by influenza viruses which belong to the Orthomyxoviridae family. Type A influenza viruses are the most important as they cause severe epidemics and are responsible of important pathological troubles. Type A influenza viruses are classified in different sub-types depending of the nature of their surface glycoproteins: haemagglutinin (H) and neuraminidase (N). The nature of the genome and the mode of replication of influenza viruses account for the high variability of these two proteins which are responsible for the immunity to the virus. The continuous appearance of point mutations in the gene coding for the H protein, leads to the progressive emergence of new viral strains. This event which is called antigenic drift makes it necessary to annually assess the composition of the human flue vaccine. Genetic reassortment is another mechanism of antigenic variation. When the gene coding for the H protein, or when both genes coding for H and N proteins are involved in genetic reassortment, a new viral sub-type occurs which replace the precedent. This event, which is termed antigenic shift, occurs occasionally every 10 to 30 years, and it is responsible of the great human pandemics. The role of the animals and particularly the importance of pigs and poultry in the emergence of these new viruses is discussed. PMID- 10687245 TI - [The artificial lens: the folly of yesterday, the standard of today]. AB - Traditional cataract operation (dislocation of the crystalline lens into the vitreous body by external pressure or by needling) is nearly as old as mankind. The idea of implanting an artificial lenticulus is not new but this was only performed for the first time in 1949. In the last 50 years, a better knowledge of corneal and retinal physiology, as well as the improvements of intraocular lens material and design, have led to very safe routine surgery. PMID- 10687246 TI - [The Faculty of Medicine and the medical practitioner in the 21st century]. PMID- 10687247 TI - [Kashin-Beck disease in China: osteochondrodysplasia related to nutrition and environment]. AB - Kashin-Beck disease is a endemic juvenile osteochondrodysplasy, whose association with selenium deficiency and/or mycotoxin toxicity has been corroborated by epidemiological studies in China, including Tibet. Iodine deficiency appears to be a new etiological factor. Together with the geographical and epidemiological exploration of the disease, scientific multidisciplinary investigations (clinics, radiological imaging, histology, environmental and molecular biology) should afford to understand the cause of the disease before it disappears as a consequence of the evolution of the Chinese Society, including in Tibet. PMID- 10687248 TI - [The program for the elimination of leprosy: ambitious challenges, novel approaches, issues in doubt]. AB - The WHO-sponsored programme on the "elimination of leprosy as a public health problem by the Year 2000" has been highly successful. Over 9 million patients were treated by multiple drug therapy. The number of patients world-wide has been reduced by more than 90 per cent over the last 10 years, being at present less than 800,000. Transmission however, the ultimate goal of the programme, has not been interrupted. The number of new cases detected per year is still high. This observation raises serious questions regarding the future of the programme. PMID- 10687249 TI - [Nuclear arms and medical responsibility]. AB - The medical profession would be unable to cope with the millions of simultaneous casualities of a nuclear war. Such a war between developed countries becomes less likely, but new countries equip themselves, with nuclear weapons, using the violation of the Non-Proliferation Treaty by the great powers as an excuse. The danger of nuclear war is spreading with the consequences one can imagine. A world movement, including military experts, political, moral and scientific leaders and representative national medical groups, among them the World Medical Association, press governments to negotiate a comprehensive abolition treaty. The "Academie royale de Medecine de Belgique" joining moral prestige to an authoritative expertise, should take a stand on the problem. PMID- 10687250 TI - Common painful foot syndromes. AB - The prevalence of foot problems in the general population is 10%, and in the elderly it ranges from 53% to 95%. Proximal plantar fasciitis is the most common cause of painful feet in clinical practice, and is twice as common among women as among men. Metatarsalgia is probably the most common cause of foot pain among middle-aged women. PMID- 10687251 TI - C-reactive protein on admission as a predictor of in-hospital death in the elderly with acute myocardial infarction. AB - BACKGROUND: The mean age of patients with acute myocardial infarction is increasing and the associated in-hospital mortality is exponentially age-related. Inflammation markers have been related to cardiovascular short and long-term prognosis. The aim of this study was to evaluate the short-term prognostic value of C-reactive protein (CRP) levels on admission in the oldest segment of the patients with acute myocardial infarction. METHODS: CRP was prospectively measured on admission by immunonephelometry in 205 consecutive old women (mean age 82 +/- 5 years) with definite acute myocardial infarction; values were then related to in-hospital mortality and the causes of death. RESULTS: CRP levels ranged from 0.1 to 31.9 mg/dl and were raised in 71% of the patients. It showed no significant correlation with baseline clinical variables such as age, history of diabetes or hypertension or prior myocardial infarction, infarct location, and time from symptom onset to admission. The overall in-hospital mortality rate was 25% and rose from 15% among patients in the lower three quartiles of CRP levels (cut point 6.4 mg/dl) to 55% among those in the upper quartile (p < 0.001). By univariate logistic-regression, the odds ratio for early death was 0.84 (95% confidence interval 0.78-0.89) for every increase by 1 mg/dl of CRP, and 5.7 (95% confidence interval 2.7-11.9) for a CRP level in the upper quartile. Multivariate analysis demonstrated the independence of the relation between CRP levels and in hospital mortality (p = 0.0001). No significant differences in CRP level were found among deceased patients classified by cause of death (heart rupture 44%, pump failure 41%, comorbidity 5%). CONCLUSIONS: CRP concentration is raised in many old patients with acute myocardial infarction and seems to independently stratify patients for in-hospital mortality risk. This prognostic information may assist in providing the appropriate level and duration of close monitoring and be an additional support to evaluate the risk-benefit ratio of thrombolytic therapy in some cases. PMID- 10687252 TI - Long-term prognostic value of the stenosis of the infarct-related artery and the presence of viable myocardium in akinetic ventricular regions in infarcted patients. AB - BACKGROUND: Recent studies have reported that adequate perfusion of the infarct related artery improves survival in patients with myocardial infarction, independently of left ventricular pump function. However, it is not known whether or not this reduction in mortality is independent of myocardial viability within the infarct zone. The aim of this study was to evaluate the prognostic value of the patency of the infarct artery and the presence of myocardial viability in akinetic regions in patients with myocardial infarction. METHODS: Low-dose dobutamine echocardiography was performed in 154 patients with recent or previous myocardial infarction and known coronary anatomy. In each patient three vascular regions were defined. Each akinetic region was considered viable if function improved during dobutamine echocardiography, and irrorated by a not stenotic akinetic area-related artery if the supplying vessel had a stenosis < 75% or had been successfully revascularized within 1 month of dobutamine echocardiography. RESULTS: At follow-up of 34 +/- 14 months, 19 patients died of cardiac death. At univariate Cox analysis end-diastolic and end-systolic volumes, ejection fraction, previous myocardial infarction, regional wall motion score index, and stenosis of the akinetic area-related artery were related to mortality. At multivariate analysis, stenosis of the akinetic area-related artery remained a significant predictor of mortality (p = 0.04), with higher mortality (13/66 vs 6/88, p = 0.02) in patients with a stenotic akinetic area-related artery, without differences in ejection fraction (35 +/- 9 vs 34 +/- 10%). Mortality was lower in patients with myocardial viability if they had a not stenotic akinetic area related artery (1/43 vs 4/21, p = 0.02), while no difference was found among non viable patients, with or without stenosis of the akinetic area-related artery (5/45 vs 9/45). CONCLUSIONS: The present study confirms the prognostic role of the patency of the infarct-related artery. However, it suggests that the lower mortality in patients with a patent artery supplying akinetic infarcted regions is related to the presence of myocardial viability in these regions. PMID- 10687253 TI - Acute anterior myocardial infarction: increased dye intensity in the myocardial risk area after coronary angioplasty is associated with reduction of diastolic volumes. AB - BACKGROUND: Myocardial perfusion in the risk area during the acute phase of myocardial infarction has been extensively investigated over the last few years. The so-called "no-reflow" or "low-reflow phenomenon" (absence of myocardial perfusion despite patency of the infarct-related coronary artery) was shown to correlate with worse postinfarction remodeling, in particular when myocardial contrast echocardiography was used. The aim of this study was to determine, during routine coronary angiography performed before and after coronary angioplasty (PTCA) during the acute phase of myocardial infarction, the existence of the no-reflow phenomenon and its relation with ventricular remodeling, by evaluating the dye video density in the myocardial risk area. This confirmation by a different diagnostic technique may serve to highlight the role of myocardial perfusion as an index of prognosis in the clinical setting of acute myocardial infarction. METHODS: Twenty-six patients (23 males, 3 females, mean age 57 +/- 8.7 years) who underwent either rescue (n = 11, 42.3%) or primary PTCA, according to clinical indications, of the left anterior descending coronary artery during an acute anterior myocardial infarction and who did not have stenosis of the left circumflex or right coronary artery, were retrospectively selected from a 6 year intake. The extent of coronary stenosis was assessed using biplane quantitative coronary angiography, while end-diastolic and end-systolic volume indexes, together with regional wall motion, were computed from echocardiography performed in the first 24 hours and at 6 months. Patients were subdivided into two groups on the basis of dye video intensity in the risk area, as assessed from images obtained during left main coronary artery injections before and immediately after PTCA. It was used a subtraction technique (Group A: increased video intensity, n = 12; Group B: no change, n = 14), assuming that higher peak intensity reflects greater myocardial blood volume. Three patients in Group B with ineffective PTCA were excluded, so that the final number of considered patients was 11. RESULTS: The distribution of rescue PTCA was similar in the two groups (7 in Group A vs 3 in Group B, p = 0.13) as were clinical characteristics and therapeutic regimen. There was a significant time * group interaction for end-diastolic volumes (-4.6 +/- 23% in Group A vs +22 +/- 22% in Group B, p = 0.029), whereas end-systolic volumes showed a tendency to greater dilation in Group B (+19 +/- 28% vs +0.9 +/- 31% in Group A), although this difference was not significant (p = 0.27). No interaction was evident for increase in the vessel area (+46 +/- 12.5% in Group A vs +43.2 +/- 13.6% in Group B, p = 0.99), or for extent of regional dysfunction (+3.08 +/- 10.9 chords in Group A vs -2.5 +/- 9.5 chords in Group B, p = 0.50). CONCLUSIONS: The detection of myocardial blood volume in the risk area using dye video intensity during left main dye contrast injection, is useful to distinguish whether there is improved perfusion at the muscular level, following successful angioplasty of the infarct-related coronary artery. Lack of improved myocardial perfusion has an adverse effect on left ventricular volumes independently of coronary stenosis resolution and regional wall motion changes in the time. PMID- 10687254 TI - Increased plasma levels of fibrinogen in acute and chronic ischemic coronary syndromes. AB - BACKGROUND: The aim of this study was to evaluate the pathophysiological role of fibrinogen in patients with chronic or acute ischemic coronary syndromes on the basis of epidemiological and clinical evidences showing the importance of fibrinogen as a risk factor for cardiovascular diseases and atherosclerosis progression. METHODS: We evaluated the behavior of plasma fibrinogen in 310 hospitalized patients with 1) acute myocardial infarction (n = 98); 2) unstable angina (n = 87); 3) chronic ischemic heart disease (n = 75); and 4) in controls without myocardial ischemia (n = 50). Fibrinogen was evaluated, by using the Clauss method, on day 1 and 5 during in hospital-stay and at 6-month follow-up in patients suffering from acute myocardial infarction. RESULTS: Plasma levels of fibrinogen were higher in patients with chronic ischemic heart disease (335.3 +/- 81.2 mg/dl, p < 0.001) and especially in patients with acute myocardial infarction (454.72 +/- 69.5 mg/dl, p < 0.00001) and unstable angina (382.6 +/- 101.3 mg/dl, p < 0.00025) in comparison with controls (271.28 +/- 62.4 mg/dl). Q wave myocardial infarction showed higher levels of fibrinogen than non-Q wave (461.3 +/- 95.8 vs 422.5 +/- 71.3 mg/dl, p < 0.02). Patients with acute myocardial infarction showed a further increase in fibrinogen on day 5 in comparison with entry levels (525.88 +/- 87.3 vs 454.7 +/- 69.5 mg/dl, p < 0.00001) regardless of the fibrinolytic treatment. Patients who died (n = 6) or had severe arrhythmias (n = 4) during in-hospital stay as well as those with post infarction angina (n = 20) showed higher fibrinogen levels. CONCLUSIONS: Our results confirm the role of fibrinogen as a risk factor for ischemic heart disease, especially in patients with unstable angina and acute myocardial infarction. In the latter, elevated fibrinogen values seem also to be associated with a worsen prognosis during hospitalization. PMID- 10687255 TI - Color duplex scanning for the identification of extracranial atherosclerosis in patients with suspected coronary artery disease. AB - BACKGROUND: The presence of extracranial artery disease has been used as a predictor of coronary artery disease (CAD). The present study was conducted to test the prevalence of extracranial artery disease among patients with suspected CAD. METHODS: Among candidates for coronary arteriography, 400 consecutive patients (mean age 63 +/- 11 years, 78% males, 22% females) underwent color duplex ultrasound of carotid arteries. RESULTS: Extracranial artery disease was documented in 60 patients (15%), CAD in 309 patients (77%). Patients with extracranial artery disease were significantly older (p < 0.001), smoked a higher amount of pack-years (p < 0.001), showed a higher incidence of diabetes (p < 0.01), hypertension (p < 0.05) and CAD (p < 0.05) when compared to extracranial artery disease-free subjects. Plotting age against extracranial artery disease and CAD distribution, extracranial artery disease occurred later in life than CAD (p < 0.001). The best cut-off point of age for predicting extracranial artery disease was 68 years. Carotid angiography was performed in 114 patients after cardiac catheterization (k = 0.8044 with color duplex scanning). CONCLUSIONS: Extracranial artery disease is frequent among patients undergoing coronary arteriography. Carotid ultrasound screening is useful in older patients. PMID- 10687256 TI - Can aortocoronary and peripheral venous bypass graft patency be improved by the administration of pentoxifylline on a long-term basis? AB - BACKGROUND: Venous bypass grafts to coronary and peripheral arteries have a high rate of occlusion at follow-up. Pentoxifylline, known to reduce blood viscosity, has the potential theoretically to reduce venous graft occlusion. We tested this hypothesis in this study. METHODS: The effect of pentoxifylline (Trental) on the early and late failure of venous bypass grafts was studied on 107 patients who underwent a total of 159 aortocoronary and 55 peripheral venous bypass grafts. Pentoxifylline was initially administered in the prime of the heart-lung machine in coronary patients and intra-arterially in peripheral vascular patients. All patients were treated postoperatively with oral administration of pentoxifylline at a dose of 400 mg daily, during the 2-year follow-up period. The short- and long-term graft patency in this unit was compared to that of an identical placebo unit, composed of 105 patients (141 aortocoronary and 55 peripheral venous bypass grafts). The patients of the placebo unit had the same operative treatment but were not treated with pentoxifylline. These patients were pre- and postoperatively examined by the same methods as the patients of the pentoxifylline unit. RESULTS: The 2-year follow-up demonstrated that in cardiac patients aortocoronary graft patency was 92.5% in the pentoxifylline unit, and 80.6% in the placebo unit. Similar results were observed in peripheral vascular patients, as well as in a small group of patients with multifocal arterial disease. CONCLUSIONS: These results indicate that pentoxifylline affects favorably the patency of both aortocoronary and peripheral bypass grafts. PMID- 10687257 TI - Comparison of trimetazidine with atenolol in patients with syndrome X: effects on diastolic function and exercise tolerance. AB - BACKGROUND: Trimetazidine has been shown to improve anginal symptoms without altering hemodynamic variables in patients with coronary artery disease. The aim of this study was to compare the effect of trimetazidine and atenolol upon symptoms, resting left ventricular filling dynamics and exercise tolerance. METHODS: Sixteen patients (3 males, 13 females, mean age 62 +/- 7 years) were randomized to receive trimetazidine for 2 weeks (20 mg 3 times daily) or atenolol (100 mg daily), according to a double-blind, randomized, cross-over, placebo controlled design. At the end of each treatment period patients underwent symptom limited exercise testing, an echocardiogram and a Doppler assessment of transmitral flow pattern. Daily life anginal symptoms were annotated on a diary throughout the study. Two patients discontinued trimetazidine because of severe palpitations and only 14 patients completed the study. RESULTS: Atenolol significantly reduced the number of anginal episodes as compared to placebo or trimetazidine (0.44 +/- 0.53, 4.8 +/- 4, 2.9 +/- 4.9, p < 0.01). On atenolol, the exercise test was negative in 8 patients, but none of the patients had a negative test while on trimetazidine. Atenolol increased both time to 1 mm ST segment depression (668 +/- 213 vs 838 +/- 81 s, p < 0.05) and Doppler-derived indices of ventricular filling (E/A ratio 0.87 +/- 0.20 vs 1.21 +/- 0.26, p < 0.05). CONCLUSIONS: These results confirm the beneficial effects of atenolol in improving symptoms, exercise performance and diastolic function in syndrome X patients. Trimetazidine did not exert any significant effect on any of the analyzed variables. Since trimetazidine has been previously shown to improve myocardial ischemia in patients with overt coronary artery disease to a similar extent of beta-blockers, it is likely that other mechanisms are responsible for angina in patients with syndrome X. PMID- 10687259 TI - [Further comment on pulmonary embolism]. PMID- 10687258 TI - [Angiotensin-II receptor inhibitors in hemodialysed uremia patients with arterial hypertension: candesartan cilexitil versus losartan]. AB - BACKGROUND: The aim of this study was to evaluate, in patients with chronic renal failure in hemodialysis and arterial hypertension, the effectiveness of a new angiotensin II receptor antagonist, the candesartan cilexitil, comparing it with losartan, the first of this new class of drugs. METHODS: We have selected 128 patients with chronic renal failure (92 males and 36 females, mean age 56 +/- 6 years) and arterial hypertension, subjected to hemodialysis 3 times a week, with hemodialytic seniority of 90 +/- 10 months. The inclusion criteria in the study were given from the presence, after 15 days of pharmacological wash-out, of values of diastolic blood pressure (DBP) > or = 95 mmHg and systolic blood pressure (SBP) > or = 150 mmHg, despite a hemodialysis correctly performed. Patients were divided into two groups whether they received single blind randomized candesartan cilexitil 16 mg or losartan 50 mg at hour 8.00 for a period of 8 weeks at the end of which, after a period of pharmacological wash-out of 15 days, the drugs were administered to inverted groups for other 8 weeks. After 4 and 8 weeks of treatment an evaluation of the anti-hypertensive effectiveness by means of medical complete visit and measurement of blood pressure were made. The statistical analysis was made by means of Student's t test for paired data. RESULTS: All the patients concluded the study. After 4 weeks of treatment SBP and DBP were reduced in the group with candesartan cilexitil with regard to baseline values (SBP 151.8 +/- 6.3 vs 159.8 +/- 5.1 mmHg, p < 0.05; DBP 93.6 +/- 4.5 vs 98.1 +/- 3.7 mmHg, p < 0.05). In the losartan group (SBP 151.8 +/- 6.3 vs 158.7 +/- 5.5 mmHg, p < 0.05; DBP 93.6 +/- 4.5 vs 97.5 +/- 3.8 mmHg, p < 0.05) no significant reduction in blood pressure values was observed compared with baseline values (SBP 158.7 +/- 5.5 vs 159.8 +/- 5.1 mmHg, NS; DBP 97.5 +/- 3.8 vs 98.1 +/- 3.7 mmHg, NS). After 8 weeks of treatment in the candesartan cilexitil group (SBP 128.3 +/- 5.9 vs 159.8 +/- 5.1 mmHg, p < 0.05; DBP 81.5 +/- 4.1 vs 98.1 +/- 3.7 mmHg, p < 0.05) and in the losartan group (SBP 151.7 +/- 5.1 vs 159.8 +/- 5.1 mmHg, p < 0.05; DBP 92.7 +/- 3.9 vs 98.1 +/- 3.7 mmHg, p < 0.05) blood pressure values were reduced in the same manner as at baseline. By comparing the two drugs, candesartan cilexitil proved to have a better antihypertensive effectiveness (SBP 128.3 +/- 5.9 vs 151.7 +/- 5.1 mmHg, p < 0.05; DBP 81.5 +/- 4.1 vs 92.7 +/- 3.9 mmHg, p < 0.05). CONCLUSIONS: Our experience suggests that angiotensin II receptor antagonists may be a therapeutic remarkable option in patients with chronic renal failure in hemodialysis and arterial hypertension; the antihypertensive effect seems to be class-specific. Nevertheless, at least for our data, a better and more rapid antihypertensive results was obtained with candesartan cilexitil. PMID- 10687260 TI - [The chemoprevention of breast cancer: an evolving reality]. PMID- 10687261 TI - [The X European Cancer Conference, 12-16 September 1999, Vienna. Carcinoma of the breast and ovary]. PMID- 10687262 TI - [An angiogenesis study of Dukes' B colonic tumors]. AB - PURPOSE: To evaluate the angiogenesis in Dukes' B colon cancer. PATIENTS AND METHODS: In 60 patients (age, 39-75 years), the microvessel density and the relationship between the angiogenesis and other histologic features were retrospectively evaluated. In an ongoing prospective study, 25 patients have been enrolled to determine the possible therapeutic implications of VEGF quantitative analysis. RESULTS: The retrospective portion of this study confirms the prognostic value of the angiogenesis in terms of recurrences and survival. At present, no conclusions can be drawn from the prospective portion of the study. PMID- 10687263 TI - [The clinical usefulness of cyclosporine therapeutic monitoring]. AB - PURPOSE: To review the cyclosporine therapy indications and evaluate determine the incidence of cyclosporine blood levels outside the therapeutic range in patients under treatment, to evaluate the practical utility of the drug monitoring. PATIENTS AND METHODS: The blood concentration of monoclonal cyclosporine was monitored in 225 patients by FPIA method, using the trough concentration. RESULTS: 58% of patients had a drug blood level within the therapeutic range, 26% and 16% of them had respectively, a value below and down this range. After correcting the drug dosage, 75% and 68% of the patients of the two subgroups had a cyclosporine blood value within the therapeutic range. CONCLUSIONS: Cyclosporine is increasingly used in transplant recipients as well as in patients with other immunological diseases. Since the number of patients, with inappropriate serum levels of cyclosporine is high, therapeutic monitoring of blood levels of the drug are of paramount importance. PMID- 10687264 TI - [The effects of eradication therapy in patients with chronic atrophic gastritis and seropositivity for anti-HP antibodies and histological negativity for Helicobacter pylori]. AB - PURPOSE: The present study was undertaken to analyze both whether the elevated Helicobacter pylori levels in patients with atrophic gastritis without histologic evidence of Helicobacter pylori would be a sign of an ongoing infection and the effects of eradication on gastric atrophy. PATIENTS AND METHODS: Twenty patients (10 M e 10 F; mean age 57.25 SD 12.19) with atrophic gastritis and elevated Helicobacter pylori titers without histological evidence for Helicobacter-like organisms were included in the study. Ten patients were randomized into eradication group (Group 1) (amoxicillin at 500 mg twice a day for 14 days, metronidazole at 500 mg twice a day for 10 days and omeprazole at 20 mg twice a day for 20 days) and 10 patients were randomized into the control group (Group 2). For all subjects, serum samples and duplicate biopsy specimens (obtained endoscopically) were collected prior the study period and approximately 6 months after the therapy or the follow-up for serum samples and 8 weeks for biopsy specimens. RESULTS: In the Group 1, the Helicobacter pylori antibody titers dropped significantly in 73.39% of the patients (p < 0.0001), while in the Group 2, the antibody titers declined only in a patient who received antibiotics during the study period (p < 0.00006). In both groups, no significant improvement of atrophic gastritis was observed. CONCLUSIONS: In conclusion, in patients with atrophic gastritis, the only histological evaluation of Helicobacter-like organisms colonization in gastric biopsy specimens, appeared in our study to underestimate the true prevalence of current HP infection and the importance of the bacterium in the pathogenesis and progression of such disease. Since HP infection is often associated with an increase of proliferative index, the eradication of HP could induce a mucosal protective effect against the other carcinogen factors, although it is extremely unlikely that it can promote the regeneration of a normal gastric mucosa. PMID- 10687265 TI - [The fundamental problem of evidence-based medicine: whose "facts" and from what do the facts come?]. PMID- 10687266 TI - [The clinical use of human recombinant TSH]. AB - The recent cloning of human TSH-beta gene has allowed the production of recombinant human TSH (rhTSH) by recombinant DNA technology in mammalian cells (Chinese hamster ovary cells). Studies aimed at biochemical and biological characterization have shown that rhTSH, unlike pituitary TSH, is highly sialylated and is biological active in stimulating c-AMP accumulation in FRTL-5 cells. Phase I/II and phase III clinical studies have been performed to evaluate the safety and efficacy of rhTSH in stimulating radioactive iodine uptake in patients after total thyroidectomy for differentiated thyroid cancer. In these patients therapy with thyroid hormones is performed to replace hormone production and to suppress TSH-stimulated tumor growth. To detect residual or recurrent cancer, the therapy has to be withdrawn in order to obtain rise in endogenous TSH to perform a total body scan. rhTSH, as a source of exogenous human TSH, has been shown as an additional diagnostic tool in the follow-up of patients with thyroid cancer. Used in patients maintained on thyroid hormone suppressive therapy, rhTSH enhances the sensitivity of serum Tg testing. Although the sensitivity of scans obtained after rhTSH administration is slightly lower than that after thyroid hormone withdrawal, the use of rhTSH avoids the clinical signs and symptom of hypothyroidism and can be used in selected patients. PMID- 10687267 TI - [Medical education centers: strategies and purpose]. AB - The introduction of new didactic guidelines, for the graduate degrees in medicine and allied disciplines, is secondary to the new needs of the National Health Care System, and in part to the significant developments of science. It is not easy to meet this challenge. It is likewise not easy to channel coherently the required changes, with respect to the scientific, clinical and didactic goals. Paradoxically the same institutions that are in such great need of transformation, are also a significant part of the existing problem. In many countries, schools of medicine have developed centers for medical education that are geared toward the development and growth of students, teachers-tutors, and patients alike. Medical education has become more global, in an attempt to meet much needed communication needs, from both ends, teachers and students, as well as the recipients of care, patients. One major goal of such centers is the introduction of innovative didactic activities. There is indeed a new tendency toward the development of methodological tracks aiming at the acquisition and consolidation of a deeper and broader cultural knowledge. Amongst these initiatives there is the introduction of an evaluation of the teaching delivered, as well as the development of a multidisciplinary approach to didactics. The latter, is a prerequisite of an effective training directed toward the development of the concept of "team approach", whose ultimate goal is patient care. In Italy, at the Universita Campus Biomedico, in Rome, one of the first of such centers of medical education has been developed. Its goal is to be both a learning organization, as well as a center for both research and clinical services. PMID- 10687268 TI - [The Lynch syndrome]. AB - Lynch syndrome is a peculiar disease, accounting for 5% of the total burden of colon cancer. Characteristics of this disease are autosomal dominant transmission, early onset, and frequent right colon localization. Diagnostic criteria, aimed to collaborative studies, are based on these features (so called Amsterdam criteria). Lynch syndrome has specific biomolecular features (microsatellite instability); mismatch repair genes have been identified as responsible of this syndrome. Lynch syndrome causes high risk for extracolonic malignancies, particularly for endometrial cancer, supposed to be related to mutation of hMSH2 gene. Another feature of Lynch syndrome tumours is better survival with respect to sporadic counterpart. Genetic test allows identifying the state of mutation carriers and selects the patients to submit to screening. Endoscopic screening has been demonstrated to reduce incidence of colorectal malignancies in this syndrome. PMID- 10687269 TI - [Mefloquine and ototoxicity: a report of 3 cases]. AB - We report these cases of high-frequency sensorineural hearing loss and tinnitus, following malaria prophylaxis with mefloquine (Lariam). Only one patient had partial remission of hearing loss after suspension of the treatment. In the remaining two cases the symptomatology remained unchanged. None of the patients reported improvement of tinnitus. Our experience suggests that a routine audiologic evaluation, before and after prophylactic use of antimalarial drugs, is important to monitor potential hearing deficit. PMID- 10687270 TI - [The successes of cancer chemotherapy. The first cures of advanced-phase neoplasms]. PMID- 10687271 TI - Consumer-driven cholecystectomy: myth or reality? AB - OBJECTIVE: To discover if a consumer-driven process is fueling the high rate of cholecystectomies in the laparoscopic era. DESIGN: A 12-question survey. SETTING: A 1000-bed, university-affiliated hospital. PATIENTS: Patients admitted for elective cholecystectomies. MAIN OUTCOME MEASURES: Survey responses suggesting positive perceptions about laparoscopic cholecystectomy persuade patients to undergo gallbladder surgery. RESULTS: Patients' symptoms persisted for a mean of 20.7 months, range zero to 204. Mean time between diagnosis and surgery was ten months, range 0.1-312. Ninety-five and three-tenths percent knew their gallbladder would be removed; 1.7 percent thought it would be opened to extract stones. Ninety-one and four-tenths percent correctly identified the procedure as "laparoscopic cholecystectomy." Fifty and four-tenths percent believed they would fully recover within a week, 24.1 percent within two weeks, and 14.7 percent within two days. Eighty-three and two-tenths percent knew laparoscopic cholecystectomy required less cutting; 33.6 percent thought a laser was used. Forty-four and eight-tenths percent assumed laparoscopy was the safer method. Sixty-two and one-tenth percent were referred for surgery by family physicians; Seven percent approached surgeons themselves. When asked who first suggested the laparoscopic procedure, 43.1 percent indicated family doctor, 41.4 percent, surgeon, and three percent requested laparoscopy themselves. Twenty-four and one tenth percent who delayed surgery despite more than 12 months of symptoms indicated escalating pain as their reason for seeking surgery now; 0.9 percent cited the newer procedure. CONCLUSIONS: We found no clear evidence that consumer awareness of laparoscopy has encouraged more cholecystectomies. Misconceptions exist about the procedure's execution and safety. Education about risks, especially bile duct injury, is needed. More frequent referral by family doctors probably plays a role in the higher incidence of cholecystectomies. PMID- 10687272 TI - High volume medical web sites. AB - In 1998, 22 million individuals reported surfing the web for medical information, and this number will increase to over 30 million by 2000. Fifteen of the highest volume medical web sites are described in this paper. Sponsorship and/or ownership of the fifteen sites varied. The government sponsors one, and some are the products of well-known educational institutions. One site is supported by a consumer health organization, and the American Medical Association was in the top 15. However, the most common owners are commercial, for-profit businesses. Attributes of the ideal site were categorized, and include a robust privacy and disclosure statement with an emphasis on education and an appropriate role for advertising. The covering of Complementary and Alternative Medicine (CAM) should be in a balanced and unbiased manner. There has to be an emphasis on knowledge based evidence as opposed to testimonials, and sources should be timely and reviewed. Bibliographies of authors need to be available. Hyperlinking to other web resources is valuable, as even the largest of sites cannot come close to covering all of medicine. PMID- 10687273 TI - Methodological summaries in epidemiology: health situation analyses. PMID- 10687274 TI - New PAHO list 6/67 for tabulation of ICD-10 mortality data. PMID- 10687275 TI - Case definitions: measles and rubella. PMID- 10687276 TI - Telomere, telomerase, tumorigenesis and therapy: an overview. AB - The ends of chromosome in higher eukaryote are termed telomere. The DNAs present at that part of chromosome is called telomeric DNA. Telomeric DNA consists of tandemly repeated DNA sequences. The replication of the ends of chromosomes is not controlled by conventional DNA polymerases rather a special kind of enzyme is involved in this process. It is a ribonucleoprotein and known as telomerase. Cells in senescence stage face telomeric crisis that leads to loss of telomeric ends. Surveillance turns to procancer cells with increased telomerase activity which is a later consequence. Based on these facts a key diagnostic approach has been developed for detection of tumour. A novel therapy for tumour repression has been developed using telomerase inhibitors. However, these inhibitors are very much effective for solid tumour therapy and conceptually will not work on hematological malignancies. PMID- 10687277 TI - Effect of salt stress on nodulation and nitrogen fixation in legumes. AB - It is now well established that almost all phases of root nodule development in legumes are adversely affected by saline conditions in the rooting medium. There is also a general agreement that the rhizobia are more tolerant to salt stress than the host plant, but they show considerable strain variability in growth and survival under saline conditions. Inhibitory effect of salinity on nodulation has been attributed to decrease in rhizobial colonisation and shrinkage and lack of root hair formation. Salt stress also induces premature senescence of already formed nodules. Both N2-fixation activity and nodule respiration are inhibited sharply on exposure of plants to saline conditions. The decrease in N2-fixation has been ascribed to direct effect on nitrogenase activity or an indirect effect through decrease in leghemoglobin content, respiratory rate, malate concentrations in nodules and photosynthate availability. Salinity increases oxygen diffusion resistance in the nodules and alters their ultrastructure. Decrease in N2-fixation in nodules under salinity is also accompanied by parallel decrease in the activity of H2O2-scavenging enzymes like catalase, ascorbate peroxidase and the level of antioxidants like ascorbic acid. Nodules appear to undergo osmoregulation under saline conditions by accumulating physiologically compatible solutes like proline, sugars (pinnitol) and lactic acid. The intensity of the adverse effects of salinity on nodule functioning depends on plant species, rhizobial strain, duration of exposure to saline conditions, nature, concentration and mode of salt application. PMID- 10687278 TI - Modification of ovine luteinizing hormone subunits with SMPT and its effect on subunit recombination, immunological activity, receptor binding and steroidogenic activity. AB - The increasing use of heterobifunctional cross-linking agents in the design of defined conjugates for selective targeting and inducing immune response has prompted us to study the role of epsilon-NH2 group modification of oLH subunits, their recombination and effect on immunoreactivity, receptor binding and biological activity. The epsilon-NH2 groups of alpha oLH and beta oLH subunits were separately modified by using SMPT. The alpha oLH-SMPT modified derivatives hybridize to beta oLH. Similarly, the beta oLH-SMPT derivatives recombined with alpha oLH. The recombination was judged by gel filtration chromatography and RP HPLC analysis. The sequential modification of subunits led to progressive reduction in immunoreactivity and receptor binding activity. The modification of six or more epsilon-NH2 groups in alpha oLH although recombine fully with native beta oLH but failed to react to anti-oLH antibody. Moreover, the steroidogenic activity was also abolished. Introduction upto four SMPT groups in alpha oLH compromised immunological and biological activities but further addition of two or more SMPT groups completely abolished antibody reactivity, receptor binding and steroidogenic activity indicating the importance of later two amino groups in the receptor binding and steroidogenic activity. The present investigation clearly demonstrate that only 1:2-3 molar ratio of oLH subunits:SMPT could generate the site(s) in the subunits of the oLH that retained reasonable immunological, receptor binding and biological activity of the hormone. Therefore, this molar ratio may be used in future for the design and synthesis of bioeffective hormonotoxins. PMID- 10687279 TI - Glutathione level and its relation to radiation therapy in patients with cancer of uterine cervix. AB - Glutathione functions as an important antioxidant in the destruction of hydrogen peroxide and lipid peroxides by providing substrate for the glutathione peroxidase and also promotes the ascorbic acid. Glutathione plays a vital role in detoxification of xenobiotics, carcinogens, free radicals and maintenance of immune functions. The study was aimed to determine plasma glutathione as well as erythrocyte glutathione and glutathione peroxidase in patients with invasive cervical carcinoma (n = 30) before initiation and after completion of radiotherapy and subsequently, at the time of first three monthly follow-up visit. The levels of plasma glutathione, erythrocyte glutathione and glutathione peroxidase activity were found to be lower in all cervical cancer patients as compared to age matched normal control women. The study indicates a change in antioxidant status in relation with the glutathione system among patients with invasive carcinoma of the uterine cervix. This study also demonstrates the effect of radiation therapy on this antioxidant system. PMID- 10687280 TI - Optimization of tumour radiotherapy: Part V--Radiosensitization by 2-deoxy-D glucose and DNA ligand Hoechst-33342 in a murine tumour. AB - Radiosensitizing effects of combination of a minor groove DNA ligand, Hoechst 33342, with the glucose analogue and inhibitor of glycolysis, 2-deoxy-D-glucose (2-DG) have been investigated in Ehrlich ascites tumour (EAT) bearing mice following focal irradiation of the tumour with 60Co gamma-rays. Treatment-induced tumour growth delay and tumour free animal survival were evaluated as parameters of radiation response. Focal irradiation of the tumour with a single fraction of 10 Gy induced a moderate delay in tumour growth but did not lead to complete regression in any of the tumours. Intravenous administration of H-342 1 hr before irradiation enhanced radiation-induced growth delay in a dose dependent manner. Complete regression of the tumour was observed only at a dose of 10 mg/kg body wt, leading to a cure (tumour free survival for more than 100 days) rate of 55%. Administration of 2-DG (2 g/kg body wt; i.v.), immediately before irradiation significantly enhanced radiation-induced growth delay and resulted in a cure rate of 45%. In combination with this dose of 2-DG (2 g/kg body wt), H-342 at a lower dose (5 mg/kg body wt) significantly enhanced the cure rate to 66%. H-342 or 2-DG given alone or in combination at the doses investigated here did not show any significant effects on the unirradiated tumour. PMID- 10687281 TI - Effect of angiotensin II on liposome uptake by the rat brain in vivo. AB - Studies have been performed to assess the possibility of using small unilamellar liposomes as therapeutic carriers to the brain of hypertensive rats. Rats were made temporal hypertensive by the infusion of angiotensin II (AII; 15 micrograms in 1 ml) through their right common carotid artery. Another control group was infused with physiological saline. Free 125iodine-BSA (125I-BSA) and 125I-BSA encapsulated liposomes (average diameter approximately equal to 100 nm) were injected in the tail vein 2 min after the infusion of AII or saline. Plasma radioactivity was monitored at different times up to 15 min when the cerebral uptake of 125I-BSA was determined. While a little variation in plasma clearance pattern of liposomes in hypertensive and control group was noticed, the uptake by cerebral tissues was markedly higher in hypertensive group. Analysis of pharmacokinetic parameters in relation to cerebral uptake indicated AII induced a short term opening of the blood-brain barrier (BBB) resulting in an increased cerebral uptake. Positively charged liposomes was found to be most effective in hypertensive state. PMID- 10687282 TI - Alterations of phosphatidylinositol signal transduction pathway in hepatic mitochondria following aflatoxin B1 administration. AB - A single dose of aflatoxin B1 (7 mg/kg body wt) to male rats significantly stimulated the turnover of mitochondrial phosphoinositides 1-7 hr following its administration. The elevation of phosphatidylinositol 3,4,5-trisphosphate was most pronounced whose level continued to be moderately high even at 17 hr period. The level of diacylglycerol showed a marked increase from 4 hr till 7 hr after carcinogen treatment, whereas that of inositol 1,4,5-trisphosphate recorded an increase with a maximum at 7 hr followed by a gradual decrease to near normal level at 24 hr period. The activation of phosphatidylinositol cycle together with an activation of PI 3-kinase, whose product PIP3 is known to be involved in apoptosis might contribute to the early step in the manifestation of toxicity and/or carcinogenicity. PMID- 10687283 TI - Liposomes of terbutaline sulphate: in vitro and in vivo studies. AB - In vitro studies were conducted to understand the comparative drug diffusion pattern, across artificial membrane, of the drug and of the prepared liposomes of different liposomal membrane composition. In vivo studies were carried out to determine the extent and time-course of pulmonary tissue uptake of administered liposomes containing terbutaline sulphate(TER) on rat lungs. In vitro studies revealed that the drug released from the prepared liposomes obeys Higuchi's diffusion controlled model. Different loading doses and release patterns of drug from the liposomes can be obtained by altering the PC:CHOL ratio and incorporation of cholesterol was found to reduce permeability of the membrane. Similarly drug absorption in vivo in rat's lung following intratracheal instillation, prolonged over 12 hr by liposomal entrapment of TER. The findings of present investigation indicated that liposomally encapsulated TER can be used for pulmonary delivery for maximizing the therapeutic efficacy and reducing undesirable side effects. PMID- 10687284 TI - Butachlor is cytotoxic and clastogenic and induces apoptosis in mammalian cells. AB - The ability of butachlor to induce cytotoxicity, clastogenicity and DNA damage was assessed using Chinese hamster ovary cells (CHO), Swiss mouse embryo fibroblasts (MEF) and human peripheral blood lymphocytes. A dose and time dependent loss of viability was evident upon treatment of CHO cells with butachlor. Cell killing to an extent of 50% was observed when cells were treated with 16.2 micrograms/ml of butachlor for 24 hr or with 11.5 micrograms/ml for 48 hr. The herbicide induced micronuclei significantly in cultured lymphocytes at 24 and 48 hr of treatment suggesting that it is clastogenic. To understand the mechanism of cell death caused by butachlor, its effect on DNA strand breaks was studied in MEF. A concomitant decrease in cell viability was observed with increase in DNA strand breaks. Agarose gel electrophoresis of DNA from herbicide treated CHO cells and cytochemical staining indicate the induction of apoptosis by butachlor. PMID- 10687285 TI - Characterization and localization of estrogen and progesterone receptors of human fallopian tube. AB - The cellular distribution of estrogen and progesterone receptors (ER and PR) in the human fallopian tube was investigated by immunohistochemical localization with specific monoclonal antibodies. Nuclear immunostaining was observed. Intense PR immunostaining was seen in tissues obtained at mid cycle and luteal stages of the normal menstrual cycle. On the other hand, enhanced staining for ER was seen in early follicular phase and mid cycle. Menopausal tissues showed negligible staining for both ER and PR. The ER and PR were characterized for their molecular size, anatomical distribution and levels during the menstrual cycle and in menopause. ER protein was present throughout the cycle and also during menopause. Western blot analysis revealed two forms of ER approximately 66 kDa and a truncated from approximately 49 kDa in hFT. Presence of A [approximately 90 kDa] and B [approximately 120 kDa] isoforms of human PR was detected. Follicular and early luteal tissue possessed relatively high concentration of immunoreactive PR whereas it was almost undetectable in menopausal tissues. These results suggests that ER and PR are regulated by the changing ovarian steroid hormones. PMID- 10687286 TI - Partial cloning and sequencing of a cDNA encoding bonnet monkey (Macaca radiata) oviduct specific protein. AB - Based on the complete nucleic acid sequence of human estrogen dependent oviductal protein and deduced amino acid sequence, potential antigenic site of the protein was identified. Two oligonucleotide primers were designed to specifically amplify the region which includes this antigenic site. With the expectation that the human, and monkey oviductins would have high nucleotide sequence homology, Bonnet monkey oviduct along with endometrium was obtained on day 5, 9, 12 and 22 of the cycle. Using RT PCR correct sized PCR product was detected in oviduct taken from day 9 and 12 of the cycle. PCR product was cloned into pBluescript KS[+] and nucleic acid sequence determined. A 96% homology to human, baboon and rhesus monkey estrogen induced glycoprotein, and a 84-88% homology to other mammalian oviductal protein was noted, thus confirming the authenticity of cDNA clone for monkey fallopian tube specific protein. PMID- 10687287 TI - Malignant transformation of Syrian hamster embryo (SHE) cells in culture by malachite green: an agent of environmental importance. AB - Malachite green (MG), consisting of green crystals with a metallic lustre, is very soluble in water and is highly cytotoxic to mammalian cells in culture and also acts as a liver tumour promoter. In view of its industrial importance and possible exposure to human beings, MG poses a potential environmental health hazard. Accordingly, we have studied the effect of MG on the formation of free radicals using Electron Spin Resonance (ESR) analysis with 5,5-dimethyl-1 pyrroline N-oxide (DMPO) as a spin trapping agent. ESR analysis showed formation of reactive free radicals during exposure of MG to Syrian hamster embryo (SHE) cells. As per mechanism-based toxicology in cancer risk assessment, the chemicals that have the potential to be metabolized to active free radical species could be human cancer hazards. So, we have investigated the effect of MG on the formation of Type II and Type III morphologically transformed foci using SHE cell transformation assay. MG induced dose related transformed foci. Some of these transformed foci were taken out using selective trypsinisation and established immortal cell lines. One of these immortal cell lines was characterized extensively. This immortal cell line showed enhanced DNA synthesis in the form of BrdU incorporation, increased presence of proliferating cell nuclear antigen (PCNA), bcl-2 and p53 proteins by immunohistochemistry. When these immortal cells were injected subcutaneously into nude mice, they developed tumors which were transplantable and histopathologically sarcomas. The present studies indicate that MG could be a potential candidate for two year chemical carcinogenesis rodent bioassays. PMID- 10687288 TI - Plant beneficial effect of two strains of Proteus vulgaris isolated from tea plantations. AB - Two strains of Proteus isolated from tea plantation soil were tested for their ability to colonise the roots of gram (Cicer arietinum), bean (Phaseolus radiatus) and mung (Phaseolus mungo) using a gnotobiotic system. Seeds bacterized with the two strains grew faster and showed significant increase in root and shoot enlargement of the plants tested. The bioactive fractions obtained from the culture filtrates and separated through HPLC showed that the plant growth promoting fractions were not always fungicidal and that the insecticidal fraction which was found only in RRLJ 16 was not plant growth promoting. These results suggest that the plant growth promotion effect of the plant beneficial bacteria may not always be due to disease suppression. PMID- 10687289 TI - Detection, prevalence, purification and characterization of lecithinase of Klebsiella pneumoniae. AB - Lecithinase activity in Klebsiella is a rare trait as out of 208 strains of Klebsiella belonging to 3 species, viz. K. pneumoniae (168), K. planticola (29) and K. oxytoca (11), only 4 strains of K. pneumoniae produced lecithinase positive colonies on egg-yolk-agar. Although cell lysates of 16 K. pneumoniae yielded positive results for lecithinase assay on egg-yolk-agar, 19 strains were detected positive for lecithinase with ELISA using anti-lecithinase serum. Release of up to 52.12% cell-bound lecithinase could be achieved with polymyxin-B treatment at 100 micrograms/ml concentration. Purified lecithinase was determined to be a high molecular weight (70 kDa), crystalizable, anionic (pI, 3.5) protein. It possessed cytolytic, haemolytic and dermonecrotic activities but did not induce fluid accumulation in rabbit ileal loop or infant mouse guts. It was inactivated by boiling, trypsin and chymotrypsin treatment and alkaline pH. Serologically, it was related to lecithinase of Aeromonas caviae and phospholipase-C of Salmonella. PMID- 10687290 TI - Cystogenesis of antral follicles induced by dehydroepiandrosterone (DHEA) stimulates mast cell proliferation and maturation in the house rat (Rattus rattus) ovary. AB - Mast cell dynamics has been studied in relation to cystogenesis of ovarian follicles in the house rat. Immature rats were injected (s.c.) daily with DHEA (6.0 mg/100 g body weight) and were sacrificed on the day 8, 16 and 24 of the start of treatment. Ovarian sections of the treated rats had majority of the antral follicles undergoing atresia or in early stages of cystogenesis. Completely developed cysts were evident from the ovarian surface after 24 days of daily treatment. Treatment for 8 days resulted in significant increase in the number of alcian blue-positive ovarian mast cells. Ovaries after 16 days of DHEA treatment showed no marked change with regard to the number of total mast cells per unit area and staining characteristics. However, a significant rise in ovarian mast cell counts was recorded after 24 days of treatment and most of the cells contained safranin-positive red granules. This increase was attributed due to the increase in their number in medulla and stroma around the cystic follicles. PMID- 10687291 TI - Studies on anti-ulcer properties of Cissampelos mucronata leaf extract. AB - The methanolic extract of the leaves of C. mucronata was screened for anti-ulcer properties using animal models. On isolated guinea pig ileum the extract inhibited contractions evoked by acetylcholine, histamine and serotonin. The extract remarkably decreased the propulsive movement of gastrointestinal content. The extract exhibited significant anti-ulcer activity protecting rats from indomethacin, histamine and stress-induced ulcers. It inhibited the growth of both Gram-positive and Gram-negative microorganisms. The oral LD50 value of the extract in mice was estimated to be 8.5 +/- 0.35 g/kg. The results revealed that the plant C. mucronata has potential medicinal value as an anti-ulcer agent. PMID- 10687293 TI - [Summary of the activities of the Academy during the 1998 fiscal year. The Royal Academy of Medicine of Belgium]. PMID- 10687292 TI - Protective effect of ellagic acid on t-butyl hydroperoxide induced lipid peroxidation in isolated rat hepatocytes. AB - Ellagic acid, a plant polyphenol, showed protective effect on isolated rat hepatocytes against destruction due to lipid peroxide formation induced by t butyl hydroperoxide in vitro. Ellagic acid inhibited the generation of superoxide anions and hydroxyl radicals both in enzymic and non enzymic systems, thus providing protection against oxidative damage. PMID- 10687294 TI - [Extraordinary births or "the thigh of Jupiter in fertilization in vitro as passing through the halo of Gargamel"]. PMID- 10687295 TI - [Thyroid and parathyroid surgery under hypnosis: from fiction to clinical application]. AB - Since 1992, we have used hypnosis routinely in more than 1400 procedures in plastic surgery. Our clinical success and experience with this technique led us to test wether hypnosis using active patient collaboration, could be used as an effective adjunct to conscious intravenous sedation ("hypnosedation", (HS)) for endocrine surgery, as an alternative to general anaesthesia. On a total of 1905 cervical endocrine surgical procedures performed between 1995 and 1998, 296 thyroidectomies and 33 cervical explorations for hyperparathyroidism were conducted under HS. Conversion to GA was needed in three cases (0.9%). All patients having HS reported a very pleasant experience and had significantly less postoperative pain while analgesic use was significantly reduced in this group. Hospital stay was also significantly shorter, providing a substantial reduction of the costs of medical care. The postoperative convalescence was significantly improved after HS and full return to social or professional activity was significantly shortened. We conclude that HS is a very efficient technique that provide physiological, psychological and economic benefits to the patient. PMID- 10687297 TI - Providing mental health services for a catchment area of 20,000,000 PMID- 10687296 TI - Bookwormburrow. PMID- 10687298 TI - The works of Charles Rycroft. PMID- 10687299 TI - Psychiatrist, know from whence you came PMID- 10687300 TI - Blueprint for a global morality--the work of Robert Rapoport 1924-1996. AB - Always an idealist, in his last work Robert Rapoport argues that we can achieve a global ethic in line with the UN Charter and that the inculcation of values must- and does--start in the home. This reviewer, while in sympathy with his aims, considers that the obstacles to agreement about ethical issues also need to be set out. PMID- 10687301 TI - A red hot issue or a red herring? The scientific credibility of psychoanalysis. AB - The author charts her personal history in so far as it relates to her interest in the subject of this paper. It is basically a review article, and the author cites and comments on publications spanning the years 1895-1996. She concludes that the scientific status of psychoanalysis is indeed a red hot issue judging from the numerous publications concerned with it that have appeared over the years. At the same time the author associates herself with those who consider the issue to be a red herring, as the terms "psychoanalysis" and "science" mean different things to different people. PMID- 10687302 TI - The "Jerusalem syndrome"--fantasy and reality a survey of accounts from the 19th century to the end of the second millennium. AB - The so-called "Jerusalem Syndrome" is behavioral phenomena observed in eccentric and psychotic tourists with religious delusions. A significant number of pilgrims and tourists have been visiting the Holy City, at least since the beginning of the 19th century, including some delusionary and eccentric characters. The authors present a selection of vivid descriptive accounts of such 19th century visitors, by six local residents and writers about Jerusalem (including one psychiatrist). It should be noted that those writers already noticed and documented the so-called syndrome more than a century ago. In comparison to modern research of the phenomena in contemporary Jerusalem, a striking similarity in the narrative and the clinical picture emerges. However, based on accumulated data, the authors suggest that in most cases the religious atmosphere of the city is not the primary cause for the disorder. The psychotic visitors had set out for their journey to the Holy City already guided by a delusionary system derived from their religious belief and cultural background. PMID- 10687303 TI - Attitudes of Israeli gay students toward other minorities: an exploratory study. AB - Notwithstanding significant changes in the legal and psychiatric status of homosexuals, they are still very much victims of public prejudice. It may be asked, however, how they view other minority groups. The present study compared the political attitudes of members of a gay political party at the Hebrew University with a control group of heterosexual university students matched for demographic characteristics. Homosexual subjects showed significantly more support for left-wing parties (particularly Meretz). A significant difference was also found in the party nominated by subjects as most opposed to their own views. Homosexual subjects showed a much higher rate of opposition to Moledet (a right wing extremist party) and Shas (a religious party), whereas heterosexual subjects were more often opposed to left-wing extremist parties. Notwithstanding this finding, levels of political tolerance for the party most opposed to their own views was higher among homosexual subjects than among the control group. Whether these results are related to the minority status of homosexuals, or to the ideology of the political left is not clear, and is recommended as a topic for further research. PMID- 10687304 TI - A survey of the teaching of undergraduate psychiatry in Israel. AB - BACKGROUND: The 1960-70s saw significant advances in the teaching of psychiatry in medical schools. Subsequently there have been developments in the structure of health care and in the methods and goals of medical education. METHOD: All four departments of undergraduate teaching of psychiatry in Israel and single departments in Greece, UK and US were asked to respond to a questionnaire about the structure and content of their teaching program. RESULTS: The amount and content of preclinical and clinical teaching have not changed over the last two decades. There has not been a significant "move into the community" and the nucleus of the program, particularly in Israel, is still in-patient psychiatry. None of the departments approached are evaluating clinical skills using performance-based techniques. CONCLUSIONS: The increased awareness of the need for physicians to have knowledge and skills in psychiatry has not lead to an increase in the teaching of psychiatry to medical students. Further, the content of teaching carried out in most of the centers studied does not meet the needs of physicians, particularly those in primary care, who have to identify and manage a wide range of psychopathology. PMID- 10687305 TI - Comments on teaching psychiatry to undergraduates. PMID- 10687306 TI - Pathoetiology and prevention of NIDDM lessons from the OLETF rat. AB - The OLETF rat, a genetic model of spontaneous development of NIDDM, exhibits hyperglycemic obesity with hyperinsulinemia and insulin resistance similar to that in humans. It is still unclear whether a defect in the beta-cell proliferation per se is the primary pathogenetic event in this model rat. To clarify this matter, we used partially pancreatectomized rats as a model. Male rats of 6 weeks of age were allocated at random to two groups: 70% pancreatectomy (Px) and sham-pancreatectomy (sham). Each group was divided into 4 subgroups by the date of sacrifice after surgery. Sustained hyperglycemia was evident in the Px OLETF rats after surgery. This was associated with insufficient proliferation of beta-cells, characterized by a decrease in beta-cell labeling with 5-bromo-2' deoxyuridine in proportion to a decrease in beta-cell mass and reduction in insulin content in the remnant pancreas. Administration of nicotinamide, however, ameliorated the sustained hyperglycemia by increasing beta-cell proliferation. These findings suggest that OLETF rats have a poor capacity for proliferation of pancreatic beta-cells, and that this change may be the critical pathogenetic event prior to the onset of overt diabetes. OLETF rats following long-term caloric restriction and spontaneous exercise training show normal glucose tolerance accompanied by an increase in GIR as shown by a euglycemic clamp. Both exercise training and caloric restriction normalize the abnormalities in the pancreas such as marked hypertrophy of islets and hyperplasia of connective tissues in islets. It is particularly noteworthy that exercise training significantly elevated the beta-cell mass/body weight ratio. This evidence obtained from OLETF rats may be of value when the mechanism of diet and exercise effects on diabetic patients are considered. PMID- 10687307 TI - The role of Wilms' tumor genes. AB - The constitutional chromosomal deletion within the short arm of one copy of chromosome 11, at band p13, which often correlated with WAGR syndrome consisting of Wilms' tumor with aniridia, genitourinary malformation, and mental retardation, provided the first clue to the genetic events in the development of Wilms' tumor. WT1 gene is encoded by 10 exons, resulting in messenger RNA subject to a complex pattern of alternative splicing. WT1 gene encodes a zinc finger transcription factor, which binds to GC-rich sequences and functions as a transcriptional activator or repressor for many growth factor genes. WT 1 protein is mainly expressed in developing kidney, testis, and ovary, indicating that it is involved in the differentiation of genitourinary tissues, all thought to be the sites of origin of Wilms' tumor. The point mutation of WT1 results in Denys Drash syndrome. The other Wilms' tumor gene, WT2 at 11p15.5, is linked to Beckwith-Wiedemann syndrome. The possibility that WT1 is involved in the etiology of rhabdoid tumor of the kidney was discussed. WT1 is expressed in immortalized hematologic cells such as EBV-LCL and hematologic malignancies, but not in PBL or IL-2L. High level WT1 expression in leukemia cells and a poor prognosis are linked in patients with leukemia, making the gene a novel marker for leukemia cells. A correlated expression between WT1 and mdr-1 in vincristine resistant cells indicates a close relation with multi-drug resistance and is a promising diagnostic marker for chemoresistance in hematologic malignancies. PMID- 10687308 TI - Role of TNF ligand and receptor family in the lymphoid organogenesis defined by gene targeting. AB - The molecular basis of lymphoid organogenesis has recently been elucidated using gene-targeted mice. Mice deficient in lymphotoxin-alpha (LT alpha) lack lymph nodes and Peyer's patches. The action of LT alpha in lymphoid organogenesis is mediated mostly by the membrane form of LT by a mechanism independent of TNF receptor I (TNFR-I) or II (TNFR-II). Additionally, follicular dendritic cell (FDC) clusters or germinal centers fail to develop in the spleen of LT alpha deficient mice. Mice deficient in either TNFR-I or LT beta R also fail to develop splenic FDC clusters and germinal centers, indicating that signaling through both TNFR-I and LT beta R is required for the development of these structures. The mechanisms underlying the defective lymphoid organogenesis in LT alpha-deficient mice, together with a natural mutant strain, alymphoplasia (aly) mice, which manifest a quite similar phenotype to LT alpha-deficient mice, were investigated by generating aggregation chimeras. These studies demonstrate that LT alpha and the aly gene product together control lymphoid organogenesis with a close mechanistic relationship in their biochemical pathways through governing distinct cellular compartments; the former acting as a circulating ligand and the latter as a LT beta R-signaling molecule expressed by the stroma of the lymphoid organs. PMID- 10687309 TI - Serum marker KL-6/MUC1 for the diagnosis and management of interstitial pneumonitis. AB - Interstitial pneumonitis includes more than a hundred diseases in which alveolitis is the main manifestation of the affected lung. Symptoms such as dry cough and exertional dyspnea, fine crackles on chest auscultation, interstitial infiltrates on chest X-ray films and CT scans, respiratory function tests, and Ga 67 scintigraphy have been used for the diagnosis and the evaluation of disease activity. However, the poor prognosis of some types of interstitial pneumonitis has not been improved. We discovered a high molecular weight mucin-like antigen, designated KL-6, which is also known as MUC1. The serum level of KL-6/MUC1 was elevated in 70-100% of patients with interstitial pneumonitis, such as pulmonary fibrosis (either idiopathic or related to collagen-vascular disorders), hypersensitivity pneumonitis, sarcoidosis, and radiation pneumonitis. The levels were significantly higher in patients with active disease than in those with inactive disease. In contrast, patients with noninterstitial lung disease did not show a significant elevation of KL-6/MUC1. Furthermore, the serum KL-6/MUC1 level was found to be an early predictive marker of the therapeutic effect of high-dose corticosteroids in patients with rapidly progressing idiopathic pulmonary fibrosis. These results indicate that KL-6/MUC1 may be a useful serum marker for the diagnosis and monitoring of patients with interstitial pneumonitis. PMID- 10687310 TI - Dietary flavonoids as antioxidants in vivo: conjugated metabolites of (-) epicatechin and quercetin participate in antioxidative defense in blood plasma. AB - Flavonoids are present in mainly plant foods and have attracted much attention in relation to disease prevention. Their antioxidant activity at least partly accounts for their potential health effect, because oxidative stress leads to a variety of pathophysiological events. It is essential to know the bioavailability of flavonoids involving intestinal absorption, metabolic conversion and urinary excretion, in order to evaluate their in vivo antioxidant activity after intake. Here (-)-epicatechin and quercetin were selected as typical flavanol- and flavonol-flavonoids present in vegetables, fruits and tea. Our rat study suggests that their metabolic conversion begins in the intestinal mucosa where the activity of uridine-5'-diphosphoglucuronosyltransferase (UGT) is at its highest. Both flavonoids accumulated mostly as glucuronide and sulfate conjugates in blood plasma after oral administration. No intact quercetin was found in the circulation. However, on the oral administration of these flavonoids, the antioxidative ability of rat plasma was enhanced indicating that conjugated metabolites participate in the antioxidant defense in blood plasma. Therefore, the intake of vegetables, fruits and tea rich in flavonoids may help to prevent oxidative damages in the blood. PMID- 10687311 TI - Effects of smoking on serum lipid and lipoprotein concentrations and lecithin: cholesterol acyltransferase activity. AB - Cigarette smoking is one of the major risk factors for cardiovascular disease. The mechanism responsible for this association is still unknown. We measured the activity of lecithin: cholesterol acyltransferase (LCAT), a key factor in the esterification of plasma cholesterol and reverse cholesterol transport, and the levels of lipids and apolipoproteins in the serum of 27 cigarette smoking and 31 non-smoking (control) men. We could not find any significant difference among these parameters between the groups. Serum LCAT activity was lower in smokers, but the difference was statistically nonsignificant. We also classified the two groups in respect to their serum lipid levels as hyper- and normolipidemic, we observed that normolipidemic-smokers had lower (p < 0.05) high density lipoprotein-cholesterol (HDL-C) and HDL-ester cholesterol levels compared to the normolipidemic-nonsmokers. While there were no any significant differences between hyperlipidemic-smokers and nonsmokers with respect to any of the parameters. In the end we have got the idea that smoking seems to affect HDL-C and HDL-ester cholesterol levels in the normolipidemic-smokers group, only, Also, LCAT activity tended to be lower in smokers compared to nonsmokers. PMID- 10687312 TI - The effect of a newly developed ointment containing eicosapentaenoic acid and docosahexaenoic acid in the treatment of atopic dermatitis. AB - While various therapeutic modalities have been tried for atopic dermatitis (AD), numerous obstinate cases exist in which sufficient effects cannot be obtained. Therefore, we developed and prepared an ointment containing docosahexaenoic acid and eicosapentaenoic acid as a topical therapeutics for AD. We applied this ointment to 64 patients with AD (aged between 2 months and 29 years) who showed poor responses to conventional therapies and obtained satisfactory results. This ointment is considered a new topical preparation for AD. PMID- 10687313 TI - Vital immunohistochemical staining for a novel method of diagnosing micro-cancer. Examination of immunohistochemical staining of non-fixed fresh tissue. AB - It becomes possible to establish a novel diagnostic method for micro-cancer by modulating the signals from the lesion, if lesions can be labeled with substances which can be detected by video endoscopy. The authors have already succeeded in synthesizing indocyanine green (ICG) derivatives for a fluorescent labeling substance which emits near-infrared rays. Before the antibodies labeled by these substances can be used, it is necessary to establish a method of vital immunohistochemical staining. So, we investigated the responses of antibodies exposed to non-fixed fresh tissue specimens as a basic study on vital immunohistochemical staining. The responses of fresh esophageal and gastric specimens (biopsied or surgically resected) to immunohistochemical staining with anti-epithelial membrane antigen (EMA) antibodies under various conditions using the ABC method were examined. These tissue specimens were stained immunohistochemically, and incubated with anti-EMA antibodies for 10 and 30 minutes (esophagus), and for 60 and 120 minutes (stomach) at 37 degrees C. These results suggest that vital immunohistochemical staining is possible under optimum conditions. If an infrared fluorescent endoscopy catching this excited fluorescence can be developed, it will be possible to establish a new endoscopic diagnostic method on the basis of vital immunohistochemical staining. PMID- 10687314 TI - Molecular genetic analysis of pyridoxine-nonresponsive homocystinuric siblings with different blood methionine levels during the neonatal period. AB - Two mutations in the cystathionine beta-synthase (CBS) gene were found in two Japanese siblings with pyridoxine non-responsive homocystinuria who had different methionine levels in their blood during the neonatal period. Both patients were compound heterozygotes of two mutant alleles: one had an A-to-G transition at nucleotide 194 (A194 G) that caused a histidine-to-arginine substitution at position 65 of the protein (H65R), while the other had a G-to-A transition at nucleotide 346 (G346A) which resulted in a glycine-to-arginine substitution at position 116 of the protein (G116R). The two mutant proteins were separately expressed in Escherichia coli, and they completely lacked catalytic activity. Despite their identical genotypes and almost equal protein intake, these siblings showed different levels of blood methionine during the neonatal period, suggesting that the level of methionine in blood is determined not only by the defect in the CBS gene and protein intake, but also by the activity of other enzymes involved in methionine and homocysteine metabolism, especially during the neonatal period. Therefore, high-risk newborns who have siblings with homocystinuria, even if the level of methionine in their blood is normal in a neonatal mass screening, should be followed up and diagnosed by an assay of enzyme activity or a gene analysis so that treatment can be begun as soon as possible to prevent the development of clinical symptoms. In addition, a new, more sensitive method for the mass screening of CBS deficiency in neonates should be developed. PMID- 10687315 TI - Fluoromicroscopic detection of myc-tagged GLUT4 on the cell surface. Co localization of the translocated GLUT4 with rearranged actin by insulin treatment in CHO cells and L6 myotubes. AB - We earlier developed a novel method to detect translocation of glucose transporter type 4 (GLUT 4) directly, quantitatively and simply using c-MYC epitope-tagged GLUT4 (GLUT4myc) Kanai F, Nishioka Y, Hayashi H, Kamohara S, Todaka M, Ebina Y: J Biol Chem 268: 14523-14526, 1993). We further developed the method to visualize GLUT4myc on the cell surface++ by fluorescence microscope using a highly sensitive immunochemical detection system in tissue culture cells stably expressing GLUT4myc. The translocation of GLUT4myc was observed on stimulation with insulin in 3T3-L1 adipocytes, CHO cells and L6 myotubes stably expressing GLUT4myc. Platelet-derived growth factor (PDGF), norepinephrine and bradykinin also triggered GLUT4 translocation in CHO-GLUT4myc cells stably expressing each receptor. To observe the distribution of GLUT4 and actin after insulin treatment, double staining for GLUT4myc and actin was performed. Translocated GLUT4myc on the cell surface was co-localized with rearranged actin in CHO cells and L6 myotubes. This result suggests that a correlation exists between GLUT4 translocation and actin rearrangement. PMID- 10687316 TI - The effects of desferrioxamine on thrombus formation in injured microvessels of the rabbit ear. AB - We investigated the effects of the iron chelator desferrioxamine (DFX) on thrombus formation in the arterioles and venules of the rabbit ear chamber. Thrombi were induced by irradiation with filtered light in combination with a fluorescent dye. The occlusive thrombus formation time was significantly extended by DFX. The morphological composition of thrombi in the arterioles and venules was different. In the arterioles, the thrombi consisted of platelet aggregation, but in the venules, platelets and leukocytes accumulated on the endothelium. This suggest that hydroxyl radicals may be important mediators in this model, as DFX is known as a hydroxyl radical scavenger. Furthermore, the components of thrombi in the arterioles and venules in the skin microvascular system may be different. PMID- 10687317 TI - Effect of intermittent liver ischemia on outcome in patients with hepatocellular carcinoma on liver cirrhosis. AB - The influence on postoperative liver function of intermittent normothermic hepatic ischemia in cirrhotic patients was studied retrospectively. The mean total ischemia time was 88 (range 30-140) minutes in the hemi-hepatic occlusion group, and 68 (range 10-187) minutes in the total occlusion group. There were no operative deaths due to hepatic failure. Postoperative liver function improved within 1 week of the operation. There was no significant difference in the incidence of postoperative complications between the groups. Thus normothermic hepatic ischemia is tolerated for up to 180 minutes in the cirrhotic liver when an intermittent technique (15 minutes clamped and 5 minutes unclamped) is used. PMID- 10687318 TI - Late recurrence of acinic cell carcinoma of the parotid gland. AB - Acinic cell carcinoma of the salivary glands is a rare cancer representing a low grade malignancy. The recurrence of a tumor is sometimes encountered, usually within 5 years of initial operation. We describe an unusual recurrence after a long interval following primary surgery. In 1987, a 60-year-old woman underwent excision of a mass in the superficial lobe of the right parotid gland under the preoperative diagnosis of a benign tumor. A histologic diagnosis of acinic cell carcinoma was made by examining sections from the resected mass. The patient noted several small nodules in the right parotid region in 1995, but she did not visit our clinic until 1998 when tenderness developed. A locally recurrent tumor and cervical lymph nodes containing metastases were resected and postoperative radiotherapy was given 11 years after the first operation. At least 10 years of follow-up may be necessary for patients with acinic cell carcinoma because of slow-tumor growth. PMID- 10687319 TI - The need for ketamine. PMID- 10687320 TI - Re: Clinical experience with oral ketamine. PMID- 10687321 TI - Development and validation of the cancer fatigue scale: a brief, three dimensional, self-rating scale for assessment of fatigue in cancer patients. AB - We herein describe the development and validation of the Cancer Fatigue Scale (CFS) for assessment of fatigue in cancer patients. We designed this scale specifically to reflect the nature of fatigue experienced by cancer patients, by using factor analysis; the CFS is a 15-item scale composed of 3 subscales (physical, affective, and cognitive subscales). Three hundred seven cancer patients participated in the validation phase. Construct validity, confirmed by repeating factor analysis, was good. Convergent validity, confirmed by a correlation between CFS and a visual analogue scale for fatigue, was also shown to be good (r = 0.67, P < 0.001). The CFS had good stability (average test-retest reliability r = 0.69, P < 0.001) and good internal consistency (Cronbach's alpha coefficient for all 15 items = 0.88). The present study indicates that the CFS is a brief, valid, and feasible measure of fatigue for use with cancer patients. PMID- 10687322 TI - Is the presence of dyspnea a risk factor for morbidity in cancer patients? AB - Data collected from six home palliative care teams in Ireland were analyzed to determine the prevalence of dyspnea in the population studied and to identify factors associated with the presence of dyspnea that might impact on future care. The prevalence of mild, moderate, or severe dyspnea, as measured by the Support Team Assessment Schedule (STAS), fell from 39% at referral in 327 evaluable patients to 23%. The presence of dyspnea at referral was positively correlated with severity of patient spiritual distress (Spearman rho = 0.110, P = 0.042) and weakness (Spearman rho = 0.105, P = 0.008) at referral. In analysis of contingency tables, dyspnea was also significantly associated with low patient (chi 2 9.5, P = 0.002) and family (chi 2 50.78, P < 0.001) well-being, high staff anxiety (chi 2 4.14, P = 0.04), male sex (chi 2 8.9, P = 0.003), a diagnosis of lung cancer (chi 2 59.88, P < 0.001), and dying in hospital rather than hospice or nursing home (chi 2 18.03, P = 0.001). In adjusting for covariates using a logistic regression analysis, however, only the presence of low family well being, a diagnosis of lung cancer, and increased likelihood of a hospital death remained significantly associated with the presence of dyspnea at referral. These data suggest that the presence of dyspnea may be associated with increased family distress, which may influence place of death. PMID- 10687324 TI - The safety and efficacy of a single dose (500 mg or 1 g) of intravenous magnesium sulfate in neuropathic pain poorly responsive to strong opioid analgesics in patients with cancer. AB - Neuropathic pain may respond poorly to morphine and is often difficult to relieve. Recent attention has been drawn to the role of the N-methyl-D-aspartate (NMDA) receptor in the potentiation of neuropathic pain. Magnesium is known to block the NMDA receptor. It reduces the neuropathic pain response in animals, and attenuates postoperative pain and migraine in humans. We have examined the safety, tolerability, and efficacy of two intravenous doses of magnesium sulfate in 12 patients with neuropathic pain due to malignant infiltration of the brachial or lumbosacral plexus. The first six patients received 500 mg, the remainder 1 g. Apart from a mild feeling of warmth at the time of the injection, both doses were well tolerated. After receiving 500 mg, three patients experienced complete pain relief and two experienced partial pain relief for up to 4 hours duration; pain was unchanged in one patient. After receiving 1 g, one patient experienced complete relief and four experienced partial pain relief of similar duration; pain was unchanged in one patient. Intravenous magnesium sulfate in these doses appears to be safe and well tolerated. A useful analgesic effect may be obtained in some patients and further evaluation is warranted. PMID- 10687323 TI - Role of octreotide, scopolamine butylbromide, and hydration in symptom control of patients with inoperable bowel obstruction and nasogastric tubes: a prospective randomized trial. AB - Bowel obstruction may be an inoperable complication in patients with end-stage cancer. Scopolamine butylbromide (SB) and octreotide (OCT) have been successfully used with the aim of reducing gastrointestinal (GI) secretions to avoid placement of a nasogastric tube (NGT); however, there have been no comparative studies concerning the efficacy of these drugs. Furthermore, there is little information about the role played by parenteral hydration in symptom control of these patients. In a prospective trial that involved all 17 inoperable bowel-obstructed patients presenting to our services with a decompressive NGT, patients were randomized to OCT 0.3 mg/day or SB 60 mg/day for 3 days through a continuous subcutaneous infusion. Clinical data, survival time, and the time interval from the first diagnosis of cancer to the onset of inoperable bowel obstruction were noted. The intensity of pain, nausea, dry mouth, thirst, dyspnea, feeling of abdominal distension, and drowsiness were assessed by means of a verbal scale before starting treatment with the drugs under study (T0) and then daily for 3 days (T1, T2, T3). Moreover, daily information was collected regarding the quantity of GI secretions through the NGT, the oral intake of fluids, the quantity of parenteral hydration, and the analgesic therapy used. The NGT could be removed in all 10 home care and in 3 hospitalized patients without changing the dosage of the drugs. OCT significantly reduced the amount of GI secretions at T2 (P = 0.016) and T3 (P = 0.020). Compared to the home care patients, the hospitalized patients received significantly more parenteral hydration (P = 0.0005) and drank more fluids (P = 0.025). There was no difference in the daily thirst and dry mouth intensity in relation to the amount of parenteral hydration or the treatment provided (OCT or SB). Independent of antisecretory treatment, the patients receiving less parenteral hydration presented significantly more nausea (T0 P = 0.002; T1 P = 0.001; T2 P = 0.003; T3 P = 0.001) and drowsiness at T3 (P < 0.5). Pain relief was obtained in all 17 patients and only two patients required an increase in morphine dose at T1. All patients with inoperable malignant bowel obstruction should undergo treatment with antisecretory drugs so as to evaluate the possibility of removing the NGT. When a more rapid reduction in GI secretions is desired, OCT should be considered as the first choice drug. Parenteral hydration over 500 ml/day may reduce nausea and drowsiness. PMID- 10687325 TI - A chart review of the ordering and documentation of urine toxicology screens in a cancer center: do they influence patient management? AB - Urine toxicology screens (UTSs) may be useful in the diagnosis or monitoring of patients with established or suspected substance abuse. In the medically ill, including those with cancer, the test may help clinicians manage therapy with controlled prescription drugs. To describe the current use of UTSs in a cancer center, the medical records of 111 patients who underwent UTS were reviewed. These 111 patients were randomly selected from a group of 215 patients who underwent screening between January 1, 1990 and December 31, 1994 (a period during which over 80,000 admissions occurred). Fifty-six of the 111 patients had evidence of one or more illicit drugs, a prescription medication that had not been ordered, or alcohol; 50 patients had negative screens. The likelihood of a positive UTS was higher if the patient had human immunodeficiency virus (HIV) infection (100% versus 46.6%) or was undergoing treatment for chronic nonmalignant pain (100% versus 43.9%). Documentation of the UTS in the medical record was infrequent: 37.8% of the charts listed no reason for obtaining the test and the ordering physician could not be identified in 29% of the records. Eighty-nine percent of the records did not contain a subsequent mention of the result of the UTS. The result was more likely to lead to a documented outcome when it was positive rather than negative (14.3% versus 0%). These results suggest that UTSs are used infrequently in the tertiary care oncology center. The documentation surrounding the ordering and subsequent use of the test in patient management is unsystematic. The appropriate use and documentation of UTSs, like substance abuse issues in general, should be a focus of staff education and quality improvement efforts. PMID- 10687326 TI - Topical capsaicin in the management of HIV-associated peripheral neuropathy. AB - Distal symmetrical peripheral neuropathy (DSPN) is a particularly distressing pain syndrome associated with human immunodeficiency virus (HIV) disease. Capsaicin has been found to be effective in relieving pain associated with other neuropathic pain syndromes, and is mentioned as a possible topical adjuvant analgesic for the relief of DSPN. This multicenter, controlled, randomized, double-masked clinical trial studied patients with HIV-associated DSPN and compared measures of pain intensity, pain relief, sensory perception, quality of life, mood, and function for patients who received topical capsaicin to the corresponding measures for patients who received the vehicle only. Twenty-six subjects were enrolled in the study. At the end of 1 week, subjects receiving capsaicin tended to report higher current pain scores than did subjects receiving the vehicle (Mann-Whitney test; P = 0.042). The dropout rate was higher for the capsaicin group (67%) than for the vehicle group (18%) (chi 2 test of association; P = 0.014). There were no other statistically significant differences between the capsaicin and vehicle groups with respect to current pain, worst pain, pain relief, sensory perception, quality of life, mood, or function at study entry or at any time during the 4-week trial. These results suggest capsaicin is ineffective in relieving pain associated with HIV-associated DSPN. PMID- 10687327 TI - Characteristics of methadone maintenance patients with chronic pain. AB - Chronic pain patients who have limited access to opioids may be redirected to methadone maintenance centers for management of their pain. Unfortunately, little information exists on the incidence and characteristics of methadone maintenance patients with chronic pain. The aim of this study was to survey individuals at methadone maintenance centers in order to determine the prevalence of chronic pain and to explore differences between patients with and without pain in this treatment setting. Of 248 participants interviewed at three centers, 152 (61.3%) reported chronic pain. Compared with patients without pain, those with pain reported significantly more health problems (P < 0.001), more psychiatric disturbance (P < 0.05), more prescription and nonprescription medication use (P < 0.001), and greater belief that they were undertreated (P < 0.001); 44% of those with pain believed that opioids prescribed for their pain had led to an addiction problem. Most of the methadone maintenance patients stated that they had always required some substance (alcohol or opioids) to feel normal. These results raise many questions about chronic-pain treatment policies and resources for persons with a history of substance abuse. Further investigations are needed to define the needs of this population and to improve their access to effective pain management. PMID- 10687328 TI - Judging the effectiveness of analgesia for children and adolescents during vaso occlusive events of sickle cell disease. AB - The effectiveness of analgesia during sickle cell crisis was examined in this descriptive, exploratory study. Pain scores (using the African-American Oucher and the Adolescent Pediatric Pain Tool) and analgesics administered were examined during a 2-hour observation/interview in the hospital while children/adolescents with sickle cell disease (SCD) experienced a vaso-occlusive episode (VOE). A convenience sample of twenty-one 6- to 16-year olds with SCD was included. Evidence indicated that 15 of the 21 children in the sample were in moderate to severe pain during their interviews, indicating that the analgesics did not effectively control their pain. Most participants (17) had received nalbuphine as the primary analgesic by intravenous infusion drip and/or patient-controlled analgesia pump. Many reasons were identified for the inadequate analgesia. The results suggested that the pain of SCD is very complex, requiring continuous adjustment of comfort measures, especially analgesics. More research is needed to examine pain control in children with SCD. PMID- 10687329 TI - Subcutaneous emphysema in advanced cancer. AB - Spontaneous subcutaneous emphysema is a rare and usually benign entity that may occasionally be symptomatic. We report a case of a patient with advanced cancer who developed extensive but asymptomatic subcutaneous emphysema shortly before death. Perforation of the lower gastrointestinal tract, perhaps as a result of straining due to severe unrelieved constipation or due to fistula formation, is suspected to have been the mechanism. The causes, treatment, and implications for management of this condition are discussed. PMID- 10687330 TI - NMDA-Receptor Antagonists: Evolving Role in Analgesia. Proceedings of a meeting. New York City, New York, USA. May 1, 1999. PMID- 10687331 TI - Update on the neurophysiology of pain transmission and modulation: focus on the NMDA-receptor. AB - Pain is detected by two different types of peripheral nociceptor neurons, C-fiber nociceptors with slowly conducting unmyelinated axons, and A-delta nociceptors with thinly myelinated axons. During inflammation, nociceptors become sensitized, discharge spontaneously, and produce ongoing pain. Prolonged firing of C-fiber nociceptors causes release of glutamate which acts on N-methyl-D-aspartate (NMDA) receptors in the spinal cord. Activation of NMDA receptors causes the spinal cord neuron to become more responsive to all of its inputs, resulting in central sensitization. NMDA-receptor antagonists, such as dextromethorphan, can suppress central sensitization in experimental animals. NMDA-receptor activation not only increases the cell's response to pain stimuli, it also decrease neuronal sensitivity to opioid receptor agonists. In addition to preventing central sensitization, co-administration of NMDA-receptor antagonists with an opioid may prevent tolerance to opioid analgesia. PMID- 10687332 TI - NMDA-receptor antagonists and opioid receptor interactions as related to analgesia and tolerance. AB - A model proposing that N-methyl-D-aspartate (NMDA) receptor and opioid receptor mechanisms overlap and interact within the same dorsal horn nociceptive neurons makes several predictions. First, hyperalgesia should be associated with opioid tolerance. Second, both hyperalgesia and tolerance to opioid-analgesia should be blocked by an NMDA-receptor antagonist. Results from our laboratory and others support these predictions and point to several clinical implications. One is that, in addition to preventing tolerance and dependence, combining NMDA-receptor antagonists with both opioid and nonopioid analgesics may increase their analgesic potency. Preclinical animal studies demonstrate these advantages and underscore the practicality of the combined administration of nontoxic NMDA receptor antagonists with various types of analgesic drugs. PMID- 10687333 TI - Chronic pain: challenges in the assessment and management of cancer pain. AB - Assessing and managing pain while caring for the whole patient is a challenge for physicians. Barriers to pain management include clinician-, patient-, and health system-related issues. The traditional model of care is focused on disease specific treatments. If these treatments fail, the focus shifts to palliation. A new model of care integrates disease-specific treatments with palliative care and rehabilitation. This model includes prevention and treatment of suffering. An essential element of this model is evaluation of the patient's concerns about the future and fear. Treating patient pain with quality pain management and palliative care involves a holistic pain assessment and management strategy. PMID- 10687334 TI - Current pharmacotherapy of chronic pain. AB - Advances in basic and clinical research have greatly expanded the options for analgesic pharmacotherapy. There are three broad categories of analgesic medications: (1) nonopioid analgesics, which includes the nonsteroidal anti inflammatory drugs (NSAIDs), acetaminophen, dipyrone, and others; (2) a diverse group of drugs known as the "adjuvant analgesics," which are defined as "drugs that have primary indications other than pain but may be analgesic in selected circumstances;" and (3) opioid analgesics. The advent of highly selective COX-2 inhibitors has generated excitement because of the possibility that these new NSAIDs will be much safer than previous COX inhibitors. However, the cost-benefit of using these relatively more expensive drugs versus other NSAIDs plus gastro protective therapies needs to be determined. Adjuvant analgesics can be grouped into four major classes according to their use: multipurpose, neuropathic pain, musculoskeletal pain, and cancer pain. There has been a dramatic increase in the number of these drugs during the past two decades and they now play an important role in the management of chronic pain. Pain specialists are now using opioids for chronic nonmalignant pain in addition to the traditional use for acute and cancer pain. This change in practice evolved from recognition that selected patients with chronic noncancer-related pain can experience sustained analgesia and function better with these drugs, without developing an addictive disorder. The combination of opioids and other drugs, such as an N-methyl-D-aspartate receptor antagonist, may improve the balance between analgesia and adverse effects. PMID- 10687335 TI - NMDA-receptor antagonists in neuropathic pain: experimental methods to clinical trials. AB - Recent clinical data suggest that chronic pain due to nerve or soft tissue injury may result in the sensitization of the central nervous system, mediated in part by the excitatory amino acids, glutamate and aspartate. Only a handful of N methyl-D-aspartate antagonists are clinically available. These include ketamine, dextromethorphan, memantine, and amantadine, as well as three clinically used opioids (methadone, dextropropoxyphene, and ketobemidone). This review summarizes the single-dose efficacy of the first two compounds in the treatment of experimental and neuropathic pain. In all examples presented here, NMDA-receptor antagonists with affinity at the phencyclidine site have been shown to modulate pain and hyperalgesia but are limited by dose-limiting side effects. Thus, provided their therapeutic ratio is favorable, NMDA-receptor antagonists may be effective in the treatment of some types of chronic pain. PMID- 10687336 TI - Abuse potential of morphine/dextromethorphan combinations. AB - The potentiation of morphine analgesia by dextromethorphan raises the issue of whether dextromethorphan also potentiates those actions of morphine that lead to abuse. Clinical pharmacology experiments indicated that dextromethorphan does not potentiate the euphorigenic and miotic actions of morphine. Morphine suppresses the dysphoric action of dextromethorphan. A second set of experiments indicated that dextromethorphan does not alter the response to naloxone-precipitated withdrawal. In a third set of experiments, dextromethorphan did not alter the morphine-induced depression in the slope of the increase in minute volume in response to breathing increased CO2. In contrast to potentiation of analgesia, dextromethorphan does not enhance the euphorigenic, physical dependence, and respiratory depressant actions of morphine. These findings indicate that dextromethorphan does not enhance the abuse potential of morphine and that the potentiation of analgesia appeared to be selective. PMID- 10687337 TI - MorphiDex pharmacokinetic studies and single-dose analgesic efficacy studies in patients with postoperative pain. AB - MorphiDex (MS:DM), a 1:1 (weight:weight) ratio of morphine sulfate (MS) to dextromethorphan hydrobromide (DM), is under clinical development for the treatment of moderate to severe pain. The enhancement of MS analgesia by DM is due to a pharmacodynamic interaction, not an effect on MS blood levels or a pharmacokinetic interaction between MS and DM, or their metabolites. Peak blood levels were achieved at approximately 1 hour, similar to MS in solution, resulting in a rapid onset of effect. As the dose was increased, dose proportionality was linear. Food reduced the peak concentration in blood (Cmax) but not the extent of the absorption. Single-dose, double-blind, placebo controlled studies in patients suffering from moderate to severe postoperative pain after third-molar oral surgery or orthopedic surgery demonstrated a significantly greater analgesic effect for MS:DM over MS alone with an 8-hour duration of effect. MS:DM is an effective treatment of moderate to severe pain. PMID- 10687338 TI - MorphiDex (MS:DM) double-blind, multiple-dose studies in chronic pain patients. AB - Preclinical and double-blind single-dose placebo-controlled studies demonstrated that MorphiDex (MS:DM), a 1:1 ratio of morphine sulfate (MS) to dextromethorphan hydrobromide (DM), provides significantly greater analgesia than an equal dose of immediate release MS, with a faster onset, and a duration of > or = 8 h. The analgesic effect of MS:DM compared to MS was evaluated in 2 double-blind, multiple-dose studies in 321 patients with cancer and other chronic pain: a crossover study that consisted of two 2-wk periods and a 4-wk parallel study. As specified in the study protocols, patients took sufficient MS or MS:DM to achieve satisfactory pain control. In the crossover study, the MS:DM group required half as much morphine as the MS group to achieve satisfactory pain control (80 mg and 162 mg, respectively). The interval between doses and the time from the last dose of the day to the first dose of the next day were significantly longer for MS:DM compared to MS. In the parallel study, MS:DM also provided pain control at a significantly lower dose. After four weeks of treatment, the mean daily dose of MS increased, while there was little change in the MS:DM mean daily dose (P = 0.025) to maintain satisfactory pain control. More patients preferred MS:DM to run-in MS than preferred MS to run-in MS (P = 0.026). The addition of DM to MS did not increase the incidence of adverse events, which were those commonly associated with opioid use. These studies confirm that MS:DM provides satisfactory pain relief but at a significantly lower morphine daily dose. PMID- 10687339 TI - Morphine with dextromethorphan: conversion from other opioid analgesics. AB - MorphiDex, a 1:1 combination of dextromethorphan and morphine, provides satisfactory pain control at a significantly lower morphine daily dose. To determine the appropriate conversion regimens from other oral or transdermal opioid analgesics to MorphiDex (MS:DM), 592 patients with moderate to severe chronic pain requiring daily use of opioid analgesics were enrolled in this multicenter, open-label study. Patients were instructed to use MS:DM as needed to achieve satisfactory pain control. Overall, study patients took a significantly lower morphine daily dose (P = 0.0001), and a higher percentage (P = 0.0001) rated therapy with MS:DM as "very good" or "excellent" compared to their prestudy opioid. The mean daily dose of MS:DM remained level throughout a 10 month study extension period, suggesting that MS:DM may inhibit the development of tolerance to morphine. Most of the adverse events observed with MS:DM were those commonly reported with opioid therapy and were mild to moderate in severity. MS:DM appears safe and effective in treating moderate to severe chronic pain. PMID- 10687340 TI - Long-term safety of MorphiDex. AB - More than 2200 subjects were enrolled in the MorphiDex (MS:DM) development program, with a 1:1 (weight:weight) ratio of morphine sulfate (MS) to dextromethorphan hydrobromide (DM). Of the 1400 subjects exposed to MorphiDex, more than 350 subjects were treated for at least 6 months, and over 200 subjects were treated for a year or longer. The clinical population comprised an approximately equal number of men (46.2%) and women (53.8%), ranging in age from 16 to 96 years, and mostly Caucasian (91.8%). The most frequent (54.8%) daily dose of MorphiDex for subjects enrolled in the clinical program was 120 mg or less. Slow DM metabolizers took significantly lower daily doses of MorphiDex than rapid metabolizers without a significant difference in the incidence of adverse events. Plasma bromide concentrations were low and showed a wide margin of safety for both slow and rapid DM metabolizers. There were no clinically significant treatment-related changes in clinical laboratory tests, neurological examinations, or vital signs. The most common adverse events seen in the multiple dose controlled studies were nausea, dizziness, vomiting, somnolence, constipation, confusion, asthenia, headache, and pruritus. With long-term treatment, the prevalence of adverse events was greatest during the first month of MorphiDex exposure and then decreased over time. The incidence of constipation remained fairly constant over time. PMID- 10687341 TI - Enhancing opioid analgesia with NMDA-receptor antagonists: clarifying the clinical importance. A roundtable discussion. PMID- 10687342 TI - [Different anamnestic approaches in the treatment of dyspepsia. Quicker access to gastroscopy should make the diagnosis easier]. PMID- 10687343 TI - [Helicobacter pylori--the discovery still of current interest in the new millenium. Both good and bad bacteria in the stomach?]. PMID- 10687344 TI - [Significant undertreatment of high cholesterol level in ischemic heart disease]. AB - Lipid and lipoprotein analysis was performed in 127 consecutive patients with stable incapaciting angina referred by cardiologists for coronary angiography (mean waiting time, 121 days). Ninety-four per cent of the patients manifested evidence of myocardial ischaemia at exercise testing, or had had earlier myocardial infarction or earlier revascularisation with angioplasty or bypass surgery. Despite the well-known results of the large statin trials in secondary prevention (4S and CARE), only a third of the patients had LDL cholesterol levels in accordance with Swedish and European guidelines, i.e., below 3.0 mmol/L. PMID- 10687345 TI - [Diagnosis and treatment of heart failure in primary health care. Low correlation between Pro-ANF and heart failure]. AB - Three eastern Swedish primary care clinics serving a predominantly rural clientele monitored for 13 months all patients under 80 years of age with a diagnosis based on clinical signs alone of heart failure (n = 56) or suspected incipient heart failure (n = 62). Echocardiography was performed on all patients. For 64% of the former group, the putative diagnosis matched echocardiography findings. Results showed a purely diastolic disorder in one-fifth of all 118 patients, and a hemodynamically significant, hitherto unknown heart defect in about as many. Pro-ANF assays correlated poorly with manifest heart failure. Pharmacological treatments were registered, and at 6-month follow-up, 82% of patients with systolic failure were receiving ACE-inhibition. PMID- 10687346 TI - [Radioimmunotherapy is rapidly developing to clinically efficient therapy]. AB - The use of monoclonal antibodies (mabs) in cancer therapy has gained renewed interest, due to recent reports of remarkable clinical response, particularly in patients with low-grade non-Hodgkin lymphoma. Better defined and more appropriate target antigens and "humanized" mabs, reducing the risk of inducing neutralising human anti-mouse antibodies, have contributed to the improvement in results. Conjugation of mabs with various radionuclides is now being explored as a means of further enhancing clinical efficacy, the idea being to allow systemic delivery of targeted radiation to areas of disease while sparing normal tissue. Radioimmunotherapy may be administered as a single large dose of radiolabelled mabs, usually requiring haematological stem cell support, or as multiple, smaller fractions. The criteria for the selection of mabs and radionuclides are discussed in the article, as are recent clinical data and the problems and prospects of future developments in radioimmunotherapy. PMID- 10687347 TI - [About knowledge and knowledge development within general practice. General practitioners meet questions of life in their medical version]. PMID- 10687348 TI - [The special physician-patient relation in connection to chronic fatigue syndrome. Anthropologists and physicians search for a new contact-model]. PMID- 10687349 TI - [Chronic fatigue syndrome--old wine in new bottles]. PMID- 10687350 TI - [To rank with safety. A new statistical method shows the way]. PMID- 10687351 TI - [Too simplified view on euthanasia. A comment to a book review]. PMID- 10687352 TI - [The first sperm captured in a picture: on its way to the ovum or out of the ovum?]. PMID- 10687353 TI - [School health services in future. Unique children in a school for all]. PMID- 10687354 TI - [Will we ever say no? For example to the RSV prophylaxis?]. PMID- 10687355 TI - [We want to put resources for the generalists!]. PMID- 10687356 TI - [The TV report on MedAnalys. An alarming example of misleading journalism]. PMID- 10687357 TI - Medical personnel and death. AB - Increased life expectancy and increases in the number of terminally ill patients, the rapid change in healthcare technology and diagnostic protocols and treatment means that we must re-examine medico-social problems and the attitude of healthcare professionals when confronted by death and terminal illnesses. The aim of this paper is to analyse the attitudes of medical personnel towards death and terminal patients. A total of 375 (168 doctors and 207 nurses) were asked about their attitudes toward death and the terminally ill. The sample comprised 165 males and 210 females aged between 20 and 64 years (mean age 34.96, SD 7.83 years). The results showed that 51.0% of the subjects interviewed were afraid of death. Although 59.4% thought that they were personally prepared to treat and help the terminally ill and 75.3% professionally prepared for such a task 42.7% of those interviewed would prefer not to work with the terminally ill. PMID- 10687358 TI - Bloodless surgery developments accommodate patients' choice of treatment. AB - The medical and ethical challenge of treating Jehovah's Witnesses without blood transfusions is being met by dedicated professionals around the world. Jehovah's Witnesses have assisted by setting up a research department (Hospital Information Services), with relevant information available at 100 branches world wide. They have appointed over 1,300 Hospital Liaison Committees in major medical centres throughout the world which disseminate relevant medical and legal literature to obviate misunderstandings. The members of these committees receive regular training and are internationally networked to assist medical and legal professionals and to avoid unnecessary confrontations. They already list over 90,000 doctors world wide who provide bloodless medical care. Around the world more than 190 hospitals offer bloodless medicine and surgery programmes. PMID- 10687359 TI - Informed consent for psychiatric research: the case of medication-free research. AB - Informed consent for research in psychiatry raises a number of ethical and legal issues. To illustrate some of the ethical problems involved, in particular issues of patient competence and proxy consent, as well as the motives of patients to become research participants, as an example the case of so-called medication-free research in schizophrenia is discussed. It is argued that an assessment of the competence of potential research subjects to consent to participation, based on explicit criteria, is necessary. In case of substituted consent it is shown that proxy decision makers may fail to protect the interests of incompetent patients in research. Finally, researchers ought to be aware that research subjects may be motivated to participate in research because they may mistakenly believe that the research project is designed to benefit them directly. PMID- 10687360 TI - Pediatric research and the parens patriae jurisdiction in Canada and England. AB - This paper considers the issue of enrolling non-autonomous minors as participants in pediatric research in light of two high court decisions concerning the sterilization of non-autonomous individuals in Canada and England. In particular, this paper looks at the 1986 case of Re Eve in which the Supreme Court of Canada found that sterilization of such persons under the parens patriae jurisdiction can be justified only if direct, therapeutic benefit to the person has been established. It is compared to the 1987 decision in Re B (a minor), in which the English Law Lords repudiated the Canadian decision and upheld the broader, traditional interpretation of the parens patriae jurisdiction which allowed best interests as justification for the non-therapeutic sterilization of non autonomous persons. With reference to differences in the interpretations of the parens patriae jurisdiction by these two courts, it is suggested that until such time that the Canadian courts consider a case directly on point, pediatric researchers in that country must presume greater restrictions when enrolling non autonomous subjects in research protocols than in countries such as England. PMID- 10687361 TI - Alternative medicine in Israel: a case of non-regulation. AB - The state of Israel is well known for its intervention in many aspects of its citizens' lives. Yet alternative medicine, a widely demanded service pertaining to a critical aspect of human life, remains outside the confines of state intervention. During Israel's fifty years of existence not a single law has been enacted to regulate alternative medicine. It is neither recognized nor banned. The major argument of this paper is that state unconcern, depicted as the antithesis of regulation, presents in a nutshell the ideological, economic, and social dilemmas confronting state and society. Official attitudes toward alternative medicine thus mirror the structure of values and the structure of power. In each of these domains positive and negative forces are identified, those seeking change (either toward banning or toward recognition) and forces aiming at preserving the status quo. PMID- 10687362 TI - The legality of slimming advertisements in Singapore. AB - Governments in newly affluent countries like Singapore perceive a growing obesity problem, which in turn attracts the attention of numerous suppliers of slimming aids of dubious value. Advertisements relating to these appear in many cases to breach the Sale of Food Act and the Medicines Act. Under these provisions, oral and transdermal slimming aids which are not prescription drugs or on the General Sales List must have a permit if they are to be advertised, which is granted if the authorities are satisfied as to the existence of evidence for efficacy. If perceived to be harmful, however, a product can be banned and if the associated advertisement is misleading it will attract a financial penalty. Slimming aids formally excluded (like herbal preparations) can be brought under the Act by executive order and banned from sale. That this formidable array of controls is rarely applied is due to the voluntary regulatory machinery of an advertising standards authority, a non-governmental body with a parallel in many other countries. However it has limited resources. It is suggested that either the executive or the advertising authority or both could adopt more rigorous criteria for the application of the law in the interests of the public. PMID- 10687363 TI - The beginning of life--ethical perspectives. AB - In 1973 Judge Blackmun, in the celebrated case of Roe v Wade, stated that "we need not resolve the difficult question of when life begins". A quarter of a century later, there is indeed a need to attempt to resolve this question. This article sets out briefly ethical issues arising from medically-assisted procreation and analyses the concepts of life, personhood and the beginning of personhood. It is submitted that the fundamental criterion in resolving the issue is to rethink the value and dignity of human life in a technologically advanced world. Since the ultimate wisdom on the purpose of life emanates from God, the article analyses diverse theological perspectives, in particular that of Christianity and Islam. In valuing human life, the article asserts that human beings must use technology responsibly and to humankind's benefit, rather than submit to exploitation by it. PMID- 10687364 TI - The Israeli Law of Freedom of Information 1998--implications with respect to the healthcare system. AB - The right of the Israeli public to get information from state authorities has been based until recently only on Israeli Supreme Court rulings. In 1998, a Law of Freedom of Information was enacted. The new Law tries to follow the rulings of the Israeli Supreme Court regarding freedom of information, in establishing substantive statutory arrangements. PMID- 10687365 TI - Care of the mentally ill offender. A model service. AB - Caring for mentally ill offenders has been a contentious issue in almost all parts of the world. In some countries such as New South Wales in Australia, a humane approach has gradually evolved over the years. This paper describes a successful service delivery model for forensic patients in a metropolitan psychiatric hospital in Sydney. Issues relating to selection criteria, problems associated with caring for the mentally ill offender in a general psychiatric hospital environment and the integration of the forensic patients with civilian psychiatric patients are discussed. PMID- 10687366 TI - Late abortion and the European convention for human rights. AB - National abortion laws usually do not allow abortion when a foetus is independently viable, i.e. from a gestational age of about 24 weeks. Fetal anomalies, which may be a reason to seek abortion, are sometimes detected only in an advanced stage of pregnancy. National legislatures who want to allow 'late' abortion need to account for the protection the fetus may derive from the European Convention for the protection of human rights. As yet it remains unclear to what extent the fetus can in fact derive protection from the Convention, although several national abortion laws have been tested against it by the European Commission. The significance of the reports of the Commission on the question whether national legislation allowing abortion of a viable fetus is in conflict with the Convention, is explored. It is concluded that there is no European legal standard in terms of duration of pregnancy to which national legislatures are committed. PMID- 10687367 TI - The role of the artificial uterus in embryo adoption and neonatal intensive care. AB - Embryo adoption is a way of having children for couples who wish to share a pregnancy experience but have neither eggs nor sperm to realise their dream. This type of situation could occur where the wife has had a hysterectomy, while her husband has an extremely low sperm count. The donors of embryos are usually couples who have completed their families. The artificial womb will duplicate the technology of a natural womb so as to enable the child to gestate and develop physically to maturity. This ability will enable the artificial womb to be used to rescue severely premature babies who would otherwise have died and allow them to develop normally to term. Legislation will be required to regulate situations where an entire pregnancy is sustained in an artificial uterus. Clarity as to parenthood in particular will need to be regulated. PMID- 10687368 TI - The appraisal of narcotic drugs in Hungary. AB - Hungary has a serious problem, historically significant and currently of increasing size, partly due to changes in trafficking routes. This paper outlines the development of measures to counter the situation, emphasising the provision of forensic laboratory services with the aid of the United Nations Drug Control program. An attempt is made to quantify more precisely permitted amounts of a drug for personal use. PMID- 10687369 TI - Humanizing nurse-patient communication: a challenge and a commitment. AB - Nurse-patient communication has been a pre-established, unvarying, technical professional and impersonal form of manifestation, reflecting mainly the achievement of nurse's instrumental role. This situation is opposed to our nursing concept as well as to the professional values that we incorporated during our professional life, all of which emphasize the importance of the person and the patient and the meaning of the visualisation of care to a total human being. The present study aims at discussing this question, propitiating an opportunity of reflection about the necessity of humanizing this relationship in the Brazilian scenery. Thus, we focused on the importance of nurse-patient relationship not only as an essential component of nursing assistance but also as a treatment in itself. In addition, we stimulate nurses to examine the way they are taking care of human beings: as objects to be manipulated and treated or as persons that need care and comprehension. The article is concluded with the thesis that nursing is not a technical profession that manipulates knowledge and technologies but a work of valuing human beings, their freedom and dignity. PMID- 10687370 TI - Police custody detainees and forensic medicine: use of medical resources in the cell block. AB - Various Dutch reports show that medical usage among those detained in police cells is many times higher than among the average population. Many visits by forensic physicians (some 36%) are due to the fact that many police custody detainees are addicted. Also, many visits are carried out as a result of the fact that the police must frequently put into custody individuals who suffer intoxications and/or injuries (some 13%) or psychiatric problems (6%-17%). In addition, many visits are for the benefit of detainees who cannot cope with conditions during their detention (21%-30%). Only a minority of the visits are to help police custody detainees who are (chronically) ill. Because of all this, a forensic physician's work mainly consists of writing repeat prescriptions, prescribing methadone, referring detainees to other assistance services and giving advice to guards or police officers on duty. This makes forensic medicine a profession in a class of its own. PMID- 10687371 TI - Patients who deceive. PMID- 10687372 TI - Get it right the first time. PMID- 10687373 TI - How to build an unbeatable estate plan. PMID- 10687374 TI - Total temporomandibular joint replacement. Who? What? When? Where? AB - The temporomandibular joint, like other joints such as the hip and knee, can be affected by a number of conditions that may lead to joint failure, necessitating total joint replacement. The TMJ Implants, Inc., or Christensen alloplastic reconstruction prostheses, play an important role in the treatment of these conditions. Patients with severely degenerated and/or nonfunctioning joints who undergo reconstruction with the Christensen total joint replacement prosthesis experience an increase in quality of life similar to that experienced by orthopedic patients with total joint replacement of the knee or hip. PMID- 10687375 TI - Acquired allergic reaction to topical anesthesia--a case report. AB - The incidence of allergic reactions to injected local anesthesia in dentistry is rare. This case was initially suspected of being such an allergic reaction. Careful history taking and immunological techniques indicated a reaction to the topical anesthesia. PMID- 10687377 TI - An example of measurement and reporting of periodontal loss of attachment (LOA) in epidemiological studies: smoking and periodontal tissue destruction. AB - The measurement and reporting of periodontal disease in epidemiological studies can be complex, with the common indices having well-recognised shortcomings. The aim of this study was to illustrate the use of the periodontal loss of attachment (LOA) approach in investigating the association between cigarette smoking and loss of periodontal attachment in a convenience sample of adults, in order to determine whether or not smoking was a risk indicator for periodontal disease. All participants were given a detailed periodontal clinical examination in two randomly assigned contralateral diagonal quadrants, with LOA measurements made at six sites per tooth. Information was also collected on participants' socio demographic characteristics, oral hygiene practices, smoking history, and attitudes towards smoking. The 240 participants examined comprised 81 current smokers (CS), 79 former smokers (FS) and 80 nonsmokers (NS). Substantial differences and a gradient in disease existed for LOA among the three groups. CS exhibited the greatest (and NS the least) prevalence, extent, and severity of LOA. CS had more plaque and calculus than either of the other two groups, but the groups did not differ with respect to bleeding on probing. Overall, smoking was associated with the disease outcome, and this persisted after potential confounders were controlled using multivariate analysis. Although the observed differences may have been due to the self-selected nature of the sample, the gradient evident across the three smoking exposure groups suggests that smoking cessation can slow the progression of the disease. The LOA approach appears to be a versatile and informative method for recording, analysing, and presenting data on periodontitis in epidemiological studies. PMID- 10687378 TI - Dental voluntary work--a year in Uganda. PMID- 10687379 TI - Atypical migration of an impacted lower third molar. AB - This report describes the atypical migration of an impacted lower third molar tooth in a 42-year-old woman. Serial radiographs showed that, over a period of 13 years, the tooth migrated from its original disto-angular position posterior to and beneath the roots of the adjacent second molar to a more horizontal position beneath the roots of the first permanent molar. The tooth was surgically removed under general anaesthesia, with biopsy and curettage of soft tissue found in the bone posterior to the tooth along the path of migration. This pattern of tooth movement is highly unusual in an adult patient. PMID- 10687380 TI - Dental treatment of patients with neurodegenerative disease. AB - Neurodegenerative disorders are among the most challenging and devastating illnesses in medicine. A heterogeneous group of chronic and progressive diseases, they include Alzheimer's, Parkinson's, and Huntington's diseases. Dentists faced with patients affected by one of these disorders are confronted with the major problems of cognition, mobility, and behaviour, as well as of dental maintenance. While treatment of patients with progressive neurodegeneration remains daunting, increased knowledge of the aetiology and pathogenesis of these diseases has provided new opportunities and a new understanding of their treatment needs. In this article, we briefly describe the effects of ageing on the brain, and introduce two of the major neurodegenerative disorders, Alzheimer's and Parkinson's diseases. The dental problems associated with these conditions include a decrease in oral hygiene, difficulty in controlling and retaining dentures, and purposeless chewing. Medications can result in xerostomia and consequent root caries and recurrent decay. Where possible, individuals with neurodegenerative disorders should always be treated by the same dentist. PMID- 10687381 TI - Synergistic insecticidal mode of action between sesquiterpene lactones and a phototoxin, alpha-terthienyl. AB - The synergistic insecticidal action of characteristic defensive substances produced by the plant family Asteraceae was investigated under controlled laboratory conditions. Sesquiterpene lactones isolated from Asteraceae that may form, through a Michael addition process, conjugates with glutathione were administered in a meridic diet to a herbivorous insect, Manduca sexta. By administering sesquiterpenes, variable in vivo reduced glutathione levels were observed in the insect larvae. When the Asteraceae-derived photooxidant alpha terthienyl was co-administered, lipid peroxidation and larval mortality were significantly enhanced in the treated groups of insects with lowered in vivo glutathione levels. PMID- 10687382 TI - UVB/UVA radiation activates a 48 kDa myelin basic protein kinase and potentiates wound signaling in tomato leaves. AB - We investigated the effect of UV radiation on early signaling events in the response of young tomato plants (Lycopersicon esculentum) to wounding. Ultraviolet-C (< 280 nm) and UVB/UVA (280-390 nm) radiation both induced 48 kDa myelin basic protein kinase activity in leaves. The activation was associated with phosphorylation of tyrosine residues on the kinase, which is indicative of protein kinases of the mitogen-activated protein kinase family. Ultraviolet-C irradiation resulted in a strong proteinase inhibitor synthesis, as reported previously (Conconi et al., Nature 383, 826-829, 1996). Under the conditions used, UVB/UVA radiation did not induce proteinase inhibitor synthesis but resulted in a strong potentiation of systemic proteinase inhibitor synthesis in response to wounding. The UVB/UVA-irradiated plants that were subsequently wounded accumulated 2.5-4-fold higher levels of proteinase inhibitor I when compared to wounded non-irradiated plants. The potentiating effect was most prominent in the systemic unwounded leaf of a wounded plant. Levels of 12-oxo phytodienoic acid and jasmonic acid that have been well documented to increase in response to wounding were not detected in response to UVB/UVA irradiation alone. The effect of UVB/UVA radiation in potentiating plant defense signaling should be further considered as a factor that may influence the ecological balance between plants and their predators. PMID- 10687383 TI - Porphyrins and related compounds as photoactivatable insecticides. 3. Laboratory and field studies. AB - The exposure of populations of Ceratitis capitata (fruit fly), Bactrocera oleae (olive fly) and Stomoxis calcitrans (house fly) to a bait containing mumolar concentrations of porphyrin-type photosensitizers resulted in a significant accumulation of the porphyrin by the insects and a consequent development of photosensitivity upon exposure to visible light. The photoinsecticidal activity appeared to increase with increasing hydrophobicity of the porphyrin molecule: thus, the amphiphilic dicationic meso-di(cis-4N-methyl-pyridyl)-cis-diphenyl porphine (n-octanol/water partition coefficient = 20) was markedly more efficient than its tricationic analogue or the dianionic hematoporphyrin (n-octanol/water partition coefficient = 12). The observed large decrease in the acetylcholinesterase activity of the photosensitized flies suggests that the damage of the nervous system gives an important contribution to the phototoxic action of porphyrins. Studies with C. capitata indicate that the photoinsecticidal action of porphyrins can be utilized to control the population of noxious insects also in open field conditions. PMID- 10687384 TI - Vitamin B6 (pyridoxine) and its derivatives are efficient singlet oxygen quenchers and potential fungal antioxidants. AB - Vitamin B6 (pyridoxine, 1) and its derivatives: pyridoxal (2), pyridoxal 5 phosphate (3) and pyridoxamine (4) are important natural compounds involved in numerous biological functions. Pyridoxine appears to play a role in the resistance of the filamentous fungus Cercospora nicotianae to its own abundantly produced strong photosensitizer of singlet molecular oxygen (1O2), cercosporin. We measured the rate constants (kq) for the quenching of 1O2 phosphorescence by 1 4 in D2O. The respective total (physical and chemical quenching) kq values are: 5.5 x 10(7) M-1 s-1 for 1; 7.5 x 10(7) M-1 s-1 for 2, 6.2 x 10(7) M-1 s-1 for 3 and 7.5 x 10(7) M-1 s-1 for 4, all measured at pD 6.2. The quenching efficacy increased up to five times in alkaline solutions and decreased approximately 10 times in ethanol. Significant contribution to total quenching by chemical reaction(s) is suggested by the degradation of all the vitamin derivatives by 1O2, which was observed as declining absorption of the pyridoxine moiety upon aerobic irradiation of RB used to photosensitize 1O2. This photodegradation was completely stopped by azide, a known physical quencher of 1O2. The pyridoxine moiety can also function as a redox quencher for excited cercosporin by forming the cercosporin radical anion, as observed by electron paramagnetic resonance. All B6 vitamers fluoresce upon UV excitation. Compounds 1 and 4 emit fluorescence at 400 nm, compound 2 at 450 nm and compound 3 at 550 nm. The fluorescence intensity of 3 increased approximately 10 times in organic solvents such as ethanol and 1,2-propanediol compared to aqueous solutions, suggesting that fluorescence may be used to image the distribution of 1-4 in Cercospora to understand better the interactions of pyridoxine and 1O2 in the living fungus. PMID- 10687385 TI - Dihydrocercosporin singlet oxygen production and subcellular localization: a possible defense against cercosporin phototoxicity in Cercospora. AB - Fungi in the genus Cercospora produce cercosporin, a potent singlet oxygen (1O2) generating photosensitizer that plays a critical role in the ability of these fungi to parasitize plants. Although plants, mice, bacteria and many fungi are sensitive to cercosporin, Cercospora species are resistant to its toxicity. The cellular resistance of these fungi to cercosporin has been correlated with fungal cell surface reducing ability and the ability to maintain cercosporin in a chemically reduced state. As a model for reduced cercosporin we employed a reduced, acetylated derivative (hexaacetyl-dihydrocercosporin, HAC) that we tested for 1O2 production in a range of solvents. We found that as a 1O2 photosensitizer, HAC was only moderately effective in organic solvents (phi SO = 0.14-0.18) and very poor in water (phi SO = 0.02-0.04). By contrast, the 1O2 quantum yield of cercosporin itself was unaffected by solvent (phi SO = 0.84 0.97). To investigate the localization of reduced cercosporin in fungal cells, we developed a fluorescence assay using laser scanning confocal microscopy. This assay showed a uniform green fluorescence, indicative of reduced cercosporin, in the cytoplasm of hyphal cells treated with cercosporin. We hypothesize that the main protection mechanism against cercosporin phototoxicity in the fungus consists of transformation of cercosporin to a reduced state and localization of this reduced form in the aqueous compartment of the cell, thus decreasing intracellular 1O2 production to levels that can be tolerated by the fungus. In addition, we have, for the first time, directly detected 1O2 phosphorescence from fungal culture, either stained with the photosensitizer rose bengal or actively synthesizing cercosporin, demonstrating 1O2 production in vivo and from cercosporin in culture. PMID- 10687386 TI - Photoyellowing inhibition of bleached high yield pulps using novel water-soluble UV screens. AB - To address the deficiencies of benzophenone UV screens for preventing brightness reversion in high yield mechanical papers, we synthesized a new series of such materials with enhanced water solubility and compatibility with the lignocellulosic substrate. A series of 2,4-dihydroxybenzophenones (DHB) were synthesized containing various Mannich bases at the C3 position of one of its rings. They possess the UV-screening ability of o-hydroxylbenzophenones, and they also contain tertiary nitrogen atoms that may function as radical scavengers. Aqueous solutions of the hydrochloride salt of 3-(dimethylaminomethylene)-2,4 dihydroxylbenzophenone (1), when applied on bleached chemithermomechanical pulp (CTMP) sheets, were significantly more efficient in preventing photoyellowing than the original DHB applied on the sheets from ethanol-water solutions. This confirmed our original hypothesis that increasing the compatibility of the UV screen with the lignocellulosic matrix would increase its efficiency in preventing photoyellowing. Compound 1, however, was found to be somewhat more effective than its hydrochloride salt toward preventing photoyellowing. This was attributed to the synergistic action of the free tertiary aminic center attached on the molecule with its UV-screening ability. To comprehend further the various parameters that influence the photoyellowing inhibition performance of these compounds and DHB with bleached CTMP pulp fibers, a series of handsheets were prepared at different pH. The interactions of the protonated compound 1 with pulp fibers were then evaluated by studying their kinetics of absorption and desorption to and from the fiber matrix. This part of our study found that the adsorption of protonated Mannich derivatives of DHB onto pulp is most likely governed by a cation-exchange mechanism involving the cationic amine group with the sulfonic and carboxylic acid groups located on the surface of the fibers. The pH the paper sheet was made from was also found to affect profoundly the adsorption and retention characteristics of these compounds onto the lignocellulosic matrix. PMID- 10687387 TI - On the interaction of UV screens with the lignocellulosic matrix. AB - In this report, we describe our attempt to understand the photochemical interactions that occur between dihydroxybenzophenone (DHB)-based UV screens and lignin when high-yield pulps are treated with such materials. Milled wood lignin (MWL) and filter paper were used as models, and various irradiation protocols were carried out in the presence and absence of UV screens. After irradiation, the lignin and the UV screen were extracted and the products analyzed. These experiments showed that upon irradiation, fragments of MWL-containing chromophores were linked to cellulose via an acid-labile linkage. In the presence of UV screens, these reactions were minimized. Molecular weight measurements of the extracted lignin showed that the MWL is degraded upon solid-state irradiation. The samples that contained UV screens showed a reduced tendency to degrade. Using quantitative 31P NMR, it was possible to probe further the detailed structural changes that occurred in MWL during irradiation. In general, DHB-based UV screens and derivatives were found to interact actively with MWL when irradiated. PMID- 10687388 TI - End-to-end diffusion on the microsecond timescale measured with resonance energy transfer from a long-lifetime rhenium metal-ligand complex. AB - We measured the end-to-end diffusion coefficient of an alkyl chain-linked donor acceptor pair using the time-resolved frequency-domain decay of the donor. The donor was a rhenium metal-ligand complex with a mean decay time ranging from 2.1 to 7.9 microseconds in the absence of the Texas red acceptor. The decay time was used to measure the donor-to-acceptor distance distribution and the mutual diffusion coefficient. Using this long lifetime donor, it was easily possible to determine a diffusion coefficient near 2 x 10(-8) cm2/s and diffusion coefficients as low as 1.3 x 10(-9) cm2/s were measurable. Such long lifetime donors should be valuable for measuring the flexing of peptides on the microsecond timescale, domain motions of proteins and lateral diffusion in membranes. The availability of microsecond decay time luminophores now allows luminescence spectroscopy to be useful generally for studies of microsecond dynamics of biological macromolecules. PMID- 10687389 TI - Effects of cytosine methylation on pyrimidine dimer formation in DNA. AB - The relative induction of cyclobutane pyrimidine dimers (CPD) and pyrimidine (6 4)pyrimidone photoproducts ([6-4]PD) was quantified in the duplex homopolymers polydeoxyadenosine:polydeoxythymidine, polydeoxyguanosine:polydeoxycytidine and polydeoxyguanosine:polydeoxy-5-methylcytidine irradiated with UVC or UVB radiation. Cytosine methylation significantly increased the yield of cytosine (6 4)PD after irradiation with UVC light and of cytosine CPD and (6-4)PD after irradiation with UVB light. The data suggest that CPD and (6-4)PD are preferentially induced at 5-methylcytosine bases in DNA of cells exposed to sunlight and comprise a major component of the mutation spectrum leading to the initiation of sunlight-induced skin cancer. PMID- 10687390 TI - Multidimensional reaction coordinate for the excited-state H-atom transfer in perylene quinones: importance of the 7-membered ring in hypocrellins A and B. AB - The excited-state intramolecular H-atom transfer reactions of hypocrellins B and A are compared by using time-resolved absorption and fluorescence upconversion techniques. The hypocrellin B photophysics are well described by a simple model involving one ground-state species and excited-state forward and reverse H-atom transfer with a nonfluorescent excited state. We suggest that excited-state conformational changes are coupled to the H-atom transfer in hypocrellin B just as gauche/anti changes are coupled to the H-atom transfer in hypocrellin A. PMID- 10687391 TI - A photophysical and photochemical study of 6-methoxy-2-naphthylacetic acid, the major metabolite of the phototoxic nonsteroidal antiinflammatory drug nabumetone. AB - Nabumetone is a phototoxic nonsteroidal antiinflammatory drug used for the treatment of osteoarthritis. However, nabumetone is considered a prodrug with its metabolite 6-methoxy-2-naphthylacetic acid the active form. Photophysical and photochemical studies on this metabolite have been undertaken. It undergoes photodecarboxylation in aerated aqueous and organic solvents. In addition to the accepted photodegradation pathway for related molecules, a new mechanism that implies generation of the naphthalene radical cation from the excited singlet and addition of O2 prior to the decarboxylation process has been demonstrated. Evidence for the involvement of the excited singlet state in this mechanism have been obtained by steady-state and time-resolved fluorescence experiments. The fluorescence quenching by O2 and the shorter singlet lifetime in aerated solvents support this assignment. Laser flash photolysis also supports this mechanism by showing the noninvolvement of the triplet in the formation of the naphthalene radical cation. Finally, the well-known electron acceptor CCl4 acts as an efficient singlet quencher, enhancing the route leading to the radical cation, preventing intersystem crossing to the triplet and thus resulting in a dramatic increase in the yield of 6-methoxy-2-naphthaldehyde, the major oxidative decarboxylation product; this constitutes unambiguous proof in favor of the new mechanistic proposals. PMID- 10687392 TI - Time-resolved fluorescence spectra of arterial fluorescent compounds: reconstruction with the Laguerre expansion technique. AB - The time-resolved fluorescence spectra of the main arterial fluorescent compounds were retrieved using a new algorithm based on the Laguerre expansion of kernels technique. Samples of elastin, collagen and cholesterol were excited with a pulsed nitrogen laser and the emission was measured at 29 discrete wavelengths between 370 and 510 nm. The expansion of the fluorescence impulse response function on the Laguerre basis of functions was optimized to reproduce the observed fluorescence emission. Collagen lifetime (5.3 ns at 390 nm) was substantially larger than that of elastin (2.3 ns) and cholesterol (1.3 ns). Two decay components were identified in the emission decay of the compounds. For collagen, the decay components were markedly wavelength dependent and hydration dependent such that the emission decay became shorter at higher emission wavelengths and with hydration. The decay characteristics of elastin and cholesterol were relatively unchanged with wavelength and with hydration. The observed variations in the time-resolved spectra of elastin, collagen and cholesterol were consistent with the existence of several fluorophores with different emission characteristics. Because the compounds are present in different proportions in healthy and atherosclerotic arterial walls, characteristic differences in their time-resolved emission spectra could be exploited to assess optically the severity of atherosclerotic lesions. PMID- 10687393 TI - Strategies for evaluation of enveloped virus inactivation in red cell concentrates using hypericin. AB - Photodynamically induced virus inactivation appears promising in preventing transmission of enveloped virus infections in transfusible blood products. The potential for utilizing hypericin as a photosensitizer to inactivate key enveloped viruses in packed red cell concentrates (PRC) was evaluated. In addition to inactivating effectively > or = 10(6) TCID50 of human immunodeficiency virus (HIV), inactivation of bovine viral diarrhea virus (BVDV) in PRC was used as a model for hepatitis C virus to overcome the deficiency in reliable experimental systems for hepatitis C virus (HCV) inactivation. BVDV was two orders of magnitude more sensitive to inactivation by hypericin than HIV. As part of the virucidal efficacy analyses, the effects of photosensitization on hemopoietic cell lines carrying quiescent integrated HIV provirus were studied as models for evaluating virus inactivation in latently infected cells. Phorbol ester-induced virus production by these cells was effectively prevented by photosensitization with hypericin. A refinement of the illumination conditions, incorporating a monochromatic sodium light source with an emission spectrum coinciding with the absorption peak of hypericin, was highly virucidal, however, caused unacceptable levels of hemolysis. Red blood cells could be protected from phototoxic cellular damage by complexing hypericin with human serum albumin (albumin-hypericin), but the decrease in hemolysis was at the expense of virucidal efficacy. Thus, excitation of hypericin with a fluorescent source appears to be useful potentially for virus inactivation in PRC. PMID- 10687394 TI - Determinants of the apoptotic response to lysosomal photodamage. AB - Studies with mouse leukemia L1210 cells revealed that selective lysosomal photodamage caused by any of three photosensitizing agents was followed by a gradual loss of the mitochondrial membrane potential (delta psi m), release of cytochrome c into the cytosol, increased DEVDase activity (a measure of levels of caspase-3) and a limited apoptotic response. Similar effects were observed in the murine hepatoma 1c1c7 cell line. Immunofluorescence techniques employing 1c1c7 cells demonstrated the immediate release of the lysosomal enzyme cathepsin B following lysosomal photodamage. These studies suggest that the cytotoxic effects of lysosomal photodamage are initiated by released lysosomal proteases that either directly and/or indirectly activate caspases as a consequence of the induction of mitochondrial damage. PMID- 10687395 TI - The role of the p53 tumor suppressor in the response of human cells to photofrin mediated photodynamic therapy. AB - Although there is evidence that the p53 tumor suppressor plays a role in the response of some human cells to chemotherapy and radiation therapy, its role in the response of human cells to photodynamic therapy (PDT) is less clear. In order to examine the role of p53 in cellular sensitivity to PDT, we have examined the clonogenic survival of normal human fibroblasts that express wild-type p53 and immortalized Li-Fraumeni syndrome (LFS) cells that express only mutant p53, following Photofrin-mediated PDT. The LFS cells were found to be more resistant to PDT compared to normal human fibroblasts. The D37 (LFS cells)/D37 (normal human fibroblasts) was 2.8 +/- 0.3 for seven independent experiments. Although the uptake of Photofrin per cell was 1.6 +/- 0.1-fold greater in normal human fibroblast cells compared to that in LFS cells over the range of Photofrin concentrations employed, PDT treatment at equivalent cellular Photofrin levels also demonstrated an increased resistance for LFS cells compared to normal human fibroblasts. Furthermore, adenovirus-mediated transfer and expression of wild type p53 in LFS cells resulted in an increased sensitivity to PDT but no change in the uptake of Photofrin per cell. These results suggest a role for p53 in the response of human cells to PDT. Although normal human fibroblasts displayed increased levels of p53 following PDT, we did not detect apoptosis or any marked alteration in the cell cycle of GM38 cells, despite a marked loss of cell viability. In contrast, LFS cells exhibited a prolonged accumulation of cells in G2 phase and underwent apoptosis following PDT at equivalent Photofrin levels. The number of apoptotic LFS cells increased with time after PDT and correlated with the loss of cell viability. A p53-independent induction of apoptosis appears to be an important mechanism contributing to loss of clonogenic survival after PDT in LFS cells, whereas the induction of apoptosis does not appear to be an important mechanism leading to loss of cell survival in the more sensitive normal human fibroblasts following PDT at equivalent cellular Photofrin levels. PMID- 10687396 TI - Biodistribution and bioactivity of tetra-pegylated meta tetra(hydroxyphenyl)chlorin compared to native meta-tetra(hydroxyphenyl)chlorin in a rat liver tumor model. AB - It has been proposed that the construction of a photosensitizer-polymer conjugate would lead to an increased selective retention of the drug in tumor tissue resulting in an enhancement of selective tumor destruction by light in photodynamic therapy. In this study the kinetics of a tetra-pegylated derivative of meta-tetra(hydroxyphenyl)chlorin (mTHPC-PEG) were compared with those of native meta-tetra(hydroxyphenyl)chlorin (mTHPC) in a rat liver tumor model. In addition, the time course of bioactivity of both drugs was studied in normal liver tissue. Pegylation of mTHPC resulted in a two-fold increase in the plasma half-life time, a five-fold decrease in liver uptake and an increase in the tumor selectivity at early time intervals after drug administration. However, although mTHPC concentrations in liver decrease rapidly with time, mTHPC-PEG liver concentrations increased as a function of time. This led to a loss of tumor selectivity at all but the earliest time points, whereas with mTHPC tumor selectivity increased with time. For both drugs the time course of bioactivity in the liver parallels drug concentration levels with extensive necrosis after irradiation of mTHPC-PEG-sensitized liver tissue up to drug-light intervals of 120 h. It is concluded that on balance mTHPC-PEG does not appear to show any benefits over native mTHPC for the treatment of liver tumors, as normal liver tissue accumulates the compound. However, pegylation is a potentially promising strategy with an increase in tumor selectivity and reduced liver uptake if accumulation in the liver can be prevented. PMID- 10687397 TI - Proteoglycan synthesis in porcine nasal cartilage grafts following Nd:YAG (lambda = 1.32 microns) laser-mediated reshaping. AB - Mechanically deformed morphologic cartilage grafts undergo temperature-dependent stress relaxation during sustained laser irradiation resulting in stable shape changes. In this study, porcine nasal septal cartilage specimens were evaluated for viability by measuring the incorporation of Na2(35)SO4 into proteoglycan (PTG) macromolecules in whole tissue culture following laser-mediated reshaping. Synthesis rates of PTG were determined by scintillation counting lyophilized specimens and normalizing these values by total protein content. Positive controls were established by inducing chondrocyte apoptosis using prolonged exposure to nitric oxide (NO). In chondrocytes, apoptosis induced using NO resulted in significantly lower PTG synthesis rates compared to untreated native specimens. Cartilage specimens were irradiated with light emitted from a Nd:YAG laser (25 W/cm2, lambda = 1.32 microns) while recording simultaneously radiometric surface temperature, internal stress and back-scattered light intensity from a probe laser. Each specimen received one, two or three sequential laser exposures. The duration of each exposure was determined from real-time measurements of characteristic changes in back-scattered light intensity that correlate with accelerated stress relaxation. A 5 min time interval between each laser exposures allowed the cartilage specimen to return to thermal equilibrium. Average PTG synthesis rates decreased with successive laser exposures, though these were always higher than baseline rates established for NO-treated tissues, suggesting that laser-mediated cartilage reshaping acutely does not eliminate the entire population of viable chondrocytes. The reduction in PTG synthesis is correlated with the time-temperature-dependent heating profile created during laser irradiation, supporting our hypothesis that careful monitoring of laser dosimetry is required to ensure chondrocyte viability. PMID- 10687398 TI - Transmission spectra of light to the mammalian retina. AB - A simple method has been developed to determine the optical properties of the anterior segment of the intact eye. This consists of a probe that is inserted into the posterior sclera and detects light passing through the anterior segment. The probe is connected to a charge-coupled device spectrophotometer via a fiber optic bundle. It was determined that the young rat anterior segment transmits light down to 300 nm, whereas calf and rabbit eyes transmit no UVB and only part of the UVA to the posterior segment. The absorbing species in these animals is most likely NAD(P)H, which has an absorption maximum at approximately 345 nm and is associated with zeta-crystallin. A young primate anterior segment transmits almost no UV with a steep increase in transmission at CA 400 nm. Because some experiments employed a light tube that is used to illuminate the retina during vitrectomies, this method can be used to determine the transmission spectra of the anterior segment of humans in vivo. PMID- 10687399 TI - Properties of the bimodal fluorescent protein produced by Photobacterium phosphoreum. AB - A fluorescent protein isolated from the deep-sea luminous bacterium Photobacterium phosphoreum strain bmFP has been purified, cloned and sequenced. The protein is 96.5% identical in amino acid sequence to FP390, the weakly fluorescent flavoprotein encoded by the luxF gene characteristic of Photobacterium species. Similar to FP390, bmFP is a dimer of two homologous subunits binding four FMN-myristate chromophores but has the distinctive feature of emitting a bimodal fluorescence with maxima at about 488 and 517 nm, hence the name bmFP. For both bands of this fluorescence, the excitation spectrum exhibits a peak at 336 nm, not corresponding to its flavin-like absorption spectrum. Heating of bmFP in urea resulted in a decrease in the intensity of the 488 nm band along with the appearance of a new fluorescence peaking at 423 nm, partially reversible upon the removal of the urea. Upon complete denaturation, either by heat or guanidium chloride at 65 degrees C, fluorescence characteristic of both free flavin and this 423 nm species appears. It is speculated that chromophores in different states of protonation, associated with a single protein, are responsible for the unusual spectral properties of bmFP. PMID- 10687400 TI - Activities of the bimodal fluorescent protein produced by Photobacterium phosphoreum strain bmFP in the luciferase reaction in vitro. AB - The activity of the bimodal fluorescent protein (bmFP) (lambda max, 488 and 517 nm) in the in vitro luciferase reaction has been studied. The bmFP that is produced by Photobacterium phosphoreum strain bmFP is a dimer of two homologous subunits binding four riboflavin 5'-phosphate (FMN)-myristate chromophores. The addition of bmFP to the luciferase reaction in the presence of the lumazine protein prevented the lumazine protein-induced blue shift in the emission band. The bmFP reduced electrochemically serves as a substrate in the luciferase reaction in the absence of added FMN, resulting in light emission with a single maximum at about 487 nm. The bmFP was also active in lieu of FMN in the NADH/FMN oxidoreductase (flavin reductase)-luciferase coupled bioluminescence reaction in the absence of added FMN. In the coupled reaction, bioluminescence with the isolated bmFP chromophore was weaker than that with the holo-bmFP. After bmFP was used in luciferase reactions initiated either chemically or electrochemically, it was still capable of emitting bimodal fluorescence. PMID- 10687401 TI - Foundations of spatial vision: from retinal images to perceived shapes. AB - Vision is based on spatial correspondences between physically different structures--in environment, retina, brain, and perception. An examination of the correspondence between environmental surfaces and their retinal images showed that this consists of 2-dimensional 2nd-order differential structure (effectively 4th-order) associated with local surface shape, suggesting that this might be a primitive form of spatial information. Next, experiments on hyperacuities for detecting relative motion and binocular disparity among separated image features showed that spatial positions are visually specified by the surrounding optical pattern rather than by retinal coordinates, minimally affected by random image perturbations produced by 3-D object motions. Retinal image space, therefore, involves 4th-order differential structure. This primitive spatial structure constitutes information about local surface shape. PMID- 10687402 TI - Society of self: the emergence of collective properties in self-structure. AB - Using cellular automata, the authors show how mutual influences among elements of self-relevant information give rise to dynamism, differentiation, and global evaluation in self-concept. The model assumes a press for integration that promotes internally generated dynamics and enables the self-structure to operate as a self-organizing dynamical system. When this press is set at high values, the self can resist inconsistent information and reestablish equilibrium after being perturbed by such information. A weak press for integration, on the other hand, impairs self-organization tendencies, making the system vulnerable to external information. Paradoxically, external information of a random nature may enhance the emergence of a stable self-structure in an initially disordered system. The simulation results suggest that important global properties of the self reflect the operation of integration processes that are generic in complex systems. PMID- 10687403 TI - A model of response time effects in symbolic comparison. AB - A cognitive process model is developed that predicts the 3 major symbolic comparison response time effects (distance, end, and semantic congruity) found in the results of the linear syllogistic reasoning task. The model includes a simple connectionist learning component and dual evidence accumulation decision-making components. It assumes that responses can be based either on information concerning the positional difference between the presented stimulus items or on information concerning the endpoint status of each of these items. The model provides an excellent quantitative account of the mean correct response times obtained from 16 participants who performed paired comparisons of 6 ordered symbolic stimuli (3-letter names). PMID- 10687404 TI - A model of dual attitudes. AB - When an attitude changes from A1 to A2, what happens to A1? Most theories assume, at least implicitly, that the new attitude replaces the former one. The authors argue that a new attitude can override, but not replace, the old one, resulting in dual attitudes. Dual attitudes are defined as different evaluations of the same attitude object: an automatic, implicit attitude and an explicit attitude. The attitude that people endorse depends on whether they have the cognitive capacity to retrieve the explicit attitude and whether this overrides their implicit attitude. A number of literatures consistent with these hypotheses are reviewed, and the implications of the dual-attitude model for attitude theory and measurement are discussed. For example, by including only explicit measures, previous studies may have exaggerated the ease with which people change their attitudes. Even if an explicit attitude changes, an implicit attitude can remain the same. PMID- 10687405 TI - Oscillator-based memory for serial order. AB - A computational model of human memory for serial order is described (OSCillator based Associative Recall [OSCAR]). In the model, successive list items become associated to successive states of a dynamic learning-context signal. Retrieval involves reinstatement of the learning context, successive states of which cue successive recalls. The model provides an integrated account of both item memory and order memory and allows the hierarchical representation of temporal order information. The model accounts for a wide range of serial order memory data, including differential item and order memory, transposition gradients, item similarity effects, the effects of item lag and separation in judgments of relative and absolute recency, probed serial recall data, distinctiveness effects, grouping effects at various temporal resolutions, longer term memory for serial order, list length effects, and the effects of vocabulary size on serial recall. PMID- 10687406 TI - Inferring children's categorizations from sequential touching behaviors: an analytical model. AB - A child is assumed to belong to 1 of 2 classes: categorizer or noncategorizer. To determine which, 4 toy animals and 4 toy vehicles were randomly arrayed for touching for 2 min. The task was to infer whether the child was a categorizer or a noncategorizer for vehicles and similarly for animals. A model is proposed that assumes a child's sequence of touches follows one probability distribution if the child is a categorizer and another distribution if the child is a noncategorizer. The proportion of children in each category and the probability of a child being a categorizer for, say, vehicles are among the quantities that can be estimated. Data from 18-month-old children are illustrative. The model appears efficient and robust. PMID- 10687407 TI - Is causal induction based on causal power? Critique of Cheng (1997). AB - The authors empirically evaluate P. W. Cheng's (1997) power PC theory of causal induction. They reanalyze some published data taken to support the theory and show instead that the data are at variance with it. Then, they report 6 experiments in which participants evaluated the causal relationship between a fictitious chemical and DNA mutations. The power PC theory assumes that participants' estimates are based on the causal power p of a potential cause, where p is the contingency between the cause and the effect normalized by the base rate of the effect. Three of the experiments used a procedure in which causal information was presented trial by trial. For these experiments, the power PC theory was contrasted with the predictions of the probabilistic contrast model and the Rescorla-Wagner theory. For the remaining 3 experiments, a summary presentation format was employed to which only the probabilistic contrast model and the power PC theory are applicable. The power PC theory was unequivocally contradicted by the results obtained in these experiments, whereas the other 2 theories proved to be satisfactory. PMID- 10687408 TI - A note on Corballis (1997) and the genetics and evolution of handedness: developing a unified distributional model from the sex-chromosomes gene hypothesis. AB - A unified, quantitative model for sex, twin, parent, and grandparent influences on handedness is presented. Recent research modeling the evolutionary development of genetic mechanisms for the transmission of handedness on the basis of genotype fitness has appeared to lead to the conclusion that a handedness gene cannot be located on the sex chromosomes. It is shown in this article, however, that this conclusion is not of general validity. The sex-chromosomes hypothesis is developed further, and it is demonstrated that a wide-ranging, detailed, and parsimonious account of the distribution of handedness is obtained when left handedness is assumed to be associated recessively, and with low penetrance, with genetic variation located on the X chromosome. PMID- 10687409 TI - [RS3PE syndrome: are they hounds or greyhounds?]. PMID- 10687410 TI - [RS3PE syndrome or benign edematous polysynovitis in the elderly. Study of 8 cases]. AB - BACKGROUND: The RS3PE syndrome is a recently described uncommon disorder in the elderly patient; in the initial reports, it was characterized by an excellent prognosis with total remission. In contrast with rheumatoid arthritis and polymyalgia rheumatic, it has been related to HLA B7. Further works have questioned its benign nature and even that it is a new entity. Our objective was to study the clinico-biological characteristics, therapy, and clinical course in patients with RS3PE attended at our clinic. METHODS: Retrospective descriptional study of patients with RS3PE attended in our clinic from January 1992 to December 1998 following the inclusion criteria proposed by Olive et al. For all patients, the following determinations and studies were performed: complete blood count, ESR, serum biochemistry, two measurements of rheumatoid factor (RF), and X-ray. Class I HLA typing was determined in five cases. RESULTS: Eight patients were found (6 men and 2 women), with a mean age of 68.7 years. The arthritis was more common at MCP, wrists, shoulder, and PIF in hands. Four patients had tenosynovitis of the hand flexors. ESR (mean: 69.3) and RCP (mean: 49.7) were increased in most cases. Seven patients had leukocytosis. HLA B7 was positive for 4 out of five patients. All patients were treated with low-dose corticosteroids, with a mean length of 9.5 months. Three patients had relapses. During follow-up after therapy had been completed, none of the patients had other diseases. CONCLUSIONS: RS3PE syndrome involves elderly patients and its clinical course is characterized by sudden onset seronegative symmetrical polysynovitis, with pitting edema over the hand dorsum and less commonly over the pretibial region. In most cases, a marked acute-phase reaction and leukocytosis were observed. Therapy with low-dose corticosteroids is effective; sometimes however, it is necessary to add antimalarial agents. The three patients who relapsed had underlying diseases: chronic lymphocytic leukemia, Sjogren's syndrome, and liver cirrhosis, respectively. The diagnosis of RS3PE should be obtained a posteriori, after a prolonged follow-up of the patient. Atypical cases, with evolution towards other rheumatic or hematological disorders or as paraneoplastic manifestation, have been reported. PMID- 10687411 TI - [Serial assessment of the nutritional status of patients infected by the human immunodeficiency virus. Role of tumor necrosis factor/its receptors]. AB - In order to analyze the nutritional status of HIV infected patients and the involvement of the tumour necrosis factor-alpha (TNF-alpha) and its soluble receptors (sTNFRI and sTNFRII) in such an status, forty HIV infected patients, with no associated systemic opportunist infections, were prospectively followed for eight months. From each patient the following were obtained: clinical history, dietetic survey, anthropometric measurements, CD4+ T lymphocyte/mm3 count, HIV load, and serum concentration of TNF and sTNFRI and sTNFRII. Patients showed a nutritional disorder which involved mainly the fat compartment (mean tricipital skin fold 9.8 +/- 4.2 mm, that is, 65.7 +/- 27.4% of the ideal fold), associated with a hypocaloric intake (mean daily intake 1,659.5 +/- 543.0 kcal), with normal proportions of the different organic principles. Serum concentrations of TNF (87.9 +/- 79.2 vs 8.7 +/- 6.1 pg/ml, p = 0.048) and its receptors, sTNFRI (6.1 +/- 2.6 vs 1.0 +/- 0.8 pg/ml, p < 0.001) and sTNFRII (41.9 +/- 18.6 vs 6.3 +/- 3.6 pg/ml, p < 0.001) were significantly higher than those detected in a sample of ten healthy controls. No correlation was found between nutritional alterations and concentrations of TNF or its receptors, viral load, and counts of CD4+ T lymphocytes/mm3. Seventeen patients completed the follow-up period. During this period, no significant modifications in the analyzed parameters were observed: tricipital skin fold, arm circumference, serum concentrations of albumin or transferrin, concentrations of tumoral necrosis factor or its receptor and caloric intake. The conclusion is that, despite the detected nutritional alterations in the nutritional status and those in the TNF/receptor system, our data no support and interrelationship between them. PMID- 10687412 TI - [Preventable adverse drug effects at an emergency department]. AB - OBJECTIVE: To determine the incidence and evaluate the preventability of adverse drug events (ADE) associated with visits to the Emergency Department at our hospital and subsequent hospital admissions. METHODS: A six-month observational study was conducted at an Emergency Department in a University Teaching Hospital (October 15th, 1995, to April 15th, 1996). The parameters influencing the preventability were identified by means of a multivariate logistic regression analysis. RESULTS: A total of 776 ADEs (2.25%) were detected out of a total of 33,975 patients attended at the Emergency Department; 178 patients were admitted. A total of 322 cases (43.3%) were classified as preventable and were graded as mild (37.1%), moderate (32.5%), severe (27.4%), and fatal (3%). The logistic regression analysis showed that preventability was related to drugs with a narrow therapeutic index (NTI) (OR: 10.12; 95%CI: 5.36-19.07), type A ADE (OR: 4.65; 95%CI: 2.79-7.78), age > or = 65 years (OR: 3.04; 95%CI: 2.13-4.34) and self administered medication (OR: 2.2; 95%CI: 1.32-3.65). Among admitted patients, oral anticoagulants, NSAIDs, digoxin, diuretics, and insulin caused adverse events which were considered as preventable in more than 50% of cases. The errors most frequently associated with preventable ADEs included inappropriate therapy monitoring (22.5%), increased doses with NTI drugs (22.3%), absence of preventive therapy (14.3%), excessive dose according to patient's characteristics (13.4%), and inappropriate self-administered medication (10%). CONCLUSIONS: The incidence of preventable ADEs (medication errors) is high and its severity is higher than that of non-preventable ADEs. A prompt development and implementation of measures leading to avoiding prescription errors and inappropriate treatment monitoring, the factors identified as responsible for preventable ADEs, is clearly warranted. PMID- 10687413 TI - [Association of blood uric acid with other cardiovascular risk factors in the male working population in Valencia]. AB - BACKGROUND: Serum uric acid has been reported to be a risk factor for cardiovascular disease (CVD). The objective of the present work was to determine the prevalence of hyperuricemia in a large size sample of a healthy male population, as well as the association between uric acid and other cardiovascular risk factors. PATIENTS AND METHODS: A cross-sectional study was conducted in a randomly selected sample of 1,564 healthy men in Valencia (Spain), aged 20-67 years, working in the automobile industry. Serum values of uric acid, cholesterol, and glucose were obtained, as well as blood pressure and body mass index measurements. An assessment was made of socio-economic data, drug therapy, and smoking. RESULTS: The overall prevalence of hyperuricemia was 5.10%; it increased with age. A marked increase (p < 0.01) of hyperuricemic individuals was observed with increased prevalence of other cardiovascular risk factors (from 1.8% with hyperuricemia alone up to 28% among individuals with four simultaneous risk factors). By means of a multivariate logistic regression analysis, the OR of hyperuricemia associated with each factor were calculated: increased serum glucose was the variable with a stronger association (OR: 2.69; 95%CI: 1.21 5.99), obesity ranking next (OR: 2.50; 95%CI: 1.42-4.49). Statistically significant associations were also observed for increased serum cholesterol, increased blood pressure, and smoking. CONCLUSIONS: The prevalence of hyperuricemia varies with the simultaneous presence of other classical cardiovascular risk factors. Even in this healthy mediterranean population, uric acid is significantly associated with several components in the plurimetabolic syndrome. PMID- 10687414 TI - [Borderline between emergencies and hospitalization in the analysis of hospital mortality]. AB - BACKGROUND: The analysis of hospital mortality rate as a measure of care quality is usually restricted to death occurred at hospital wards, and no consideration is given to deaths occurred at the Emergency Department. Therefore, the information from a fundamental hospital area goes without analysis. METHODS: The following characteristics of decreased individuals at the Emergency Department (n = 79) and hospital wards (n = 280) in the Costa del Sol Hospital (Marbella, Malaga, Spain) during 1997 were compared: age, sex, main diagnosis at admission, main diagnosis specificity, and number of secondary diagnoses. A reevaluation of hospital mortality rates was made after data from ED deaths had been added. RESULTS: The addition of deaths occurred at the ED meant a relevant increase in hospital mortality rates: 57% for heart failure, 30% for stroke, and 25% for myocardial infarction. Twenty percent of deaths at the ED had non-specific diagnosis versus 5% at wards (p < 0.0001; 95%CI: 6.03; 24.15). Deaths at the ED had 2.9 +/- 1.3 secondary diagnoses versus 4.9 +/- 2.0 in deaths at hospital wards (p < 0.0001; 95%CI: 1.6; 2.4). CONCLUSIONS: Deaths at the ED make up a relevant proportion of the total deaths and should be incorporated to the hospital mortality analysis. Deficiencies in the collection of clinical information were observed in this ED. Therefore, adjustments for severity--an essential issue for comparing mortality rates between centers--might be precluded. PMID- 10687415 TI - [Malignant otitis externa and diabetes: report of 4 cases]. AB - Malignant otitis externa (MOE) is an uncommon infective but potentially fatal entity caused by Pseudomonas aeruginosa. It involves almost exclusively advanced aged diabetic patients. We report here four cases diagnosed at our hospital during the last 7 years. Clinical manifestations included otalgia, purulent otorrhoea, involvement of different cranial nerves and bony destruction; one patient died because of bronchoaspiration and two are alive but with sequelae. MOE should be suspected in every diabetic patient with otitis which goes unresolved with the usual antibiotic therapy. On the other hand, facial palsy should not always be attributed to a diabetic mononeuropathy and the presence of MOE should be ruled out when otitis coexists or precedes it. PMID- 10687416 TI - [Diagnostic difficulties in cystic fibrosis]. PMID- 10687417 TI - [Celiac sprue. Range of clinicobiological manifestations, current status of diagnosis, therapeutic potential, and clinical course complications]. PMID- 10687418 TI - [Current features of antihypertensive drug treatment]. PMID- 10687419 TI - [Treatment of gout]. PMID- 10687420 TI - [Long-standing skin nodular lesions in a 14-year-old male]. PMID- 10687421 TI - [Lung nodules in a patient with HIV infection and severe neutropenia]. PMID- 10687422 TI - [Challenge in the differential diagnosis of back pain]. PMID- 10687423 TI - [Weight loss, fever, and pancytopenia in a patient with skin lesions]. PMID- 10687424 TI - [Right exophthalmos]. PMID- 10687425 TI - [Clinical course risk factors in mild craniocerebral injuries (reply)]. PMID- 10687426 TI - [Tuberculosis, AIDS, and antiretroviral therapy]. PMID- 10687427 TI - [Septic abortion caused by Campylobacter jejuni]. PMID- 10687428 TI - [Thrombocytopenic thrombotic purpura, HIV, and antiviral therapy]. PMID- 10687429 TI - [Splenomegaly and peripheral lymphocytosis secondary to splenic lymphoma]. PMID- 10687430 TI - [Fatal poisoning caused by aconitine alkaloid]. PMID- 10687431 TI - [Tuberculosis mortality in the elderly]. PMID- 10687432 TI - [Arthritis and tenosynovitis caused by Mycobacterium kansasii associated with human immunodeficiency virus infection]. PMID- 10687433 TI - New drug approvals in 1998. PMID- 10687434 TI - Ethical issues in the acceptance of gifts: Part 1. AB - It brightens the day of any health care provider when a patient presents him or her with a personal gift. These thoughtful items express sincere appreciation and are evidence of a solid, trust-based provider-patient relationship. However, there is a significant ethical difference between accepting small gifts of appreciation versus gifts of great financial value. While the patient may be expressing the same appreciation with this substantial gift, he or she may also be attempting to curry special consideration or feel some pressure to please the provider. In addition, acceptance of these large gifts produces the appearance of an improper relationship between the doctor and the gift-giving patient. Patients who offer gifts of great value should receive a sensitive explanation as to why the gift cannot be accepted. PMID- 10687435 TI - The importance and use of articulators in esthetic dentistry. PMID- 10687436 TI - Esthetic improvements. PMID- 10687437 TI - The impact of current alternative herbal remedies on dental patient management. AB - With the recent boom in holistic and herbal medicine and an ever-growing trend among the general population to refer to herbal remedies as an alternative to traditional pharmaceutical therapies, dental health care providers must be aware of the wide consumption of such products and understand their nature. It becomes imperative, therefore, to include questions regarding the use of herbal preparations as a matter of routine in the patient's drug history, since this may impact a safe dental patient care delivery. PMID- 10687438 TI - Lichen planus--report of successful treatment with aloe vera. AB - Lichen planus is a disease that involves the skin and mucous membranes. It is characterized by unique eruptions. The cause of this disease is unknown, but has been linked to emotional stress, and has also been attributed to viral infections. A case is described of a successful treatment of lichen planus. PMID- 10687439 TI - Nonsurgical repair of furcal perforations: a literature review. AB - The important steps in the management of a furcal perforation are immediate action, adequate isolation, debridement, and sealing of the defect. Studies have shown that repair materials or underlying matrix material such as amalgam, Cavit, calcium hydroxide, glass ionomers, hydroxylapatite, tricalcium phosphate, and demineralized freeze-dried bone have not been able to produce consistent results. However, current research on new materials such as mineral trioxide aggregate may advance treatment modalities significantly for furcation repair. PMID- 10687440 TI - Endodontic working length determination--where does it end? AB - Obtaining a correct working length is critical to the success of endodontic therapy. Failure to identify this crucial measurement can result in untoward treatment outcomes, which may include increased patient discomfort, possible infection or cyst development, and extrusion or intra-canal medication into the periradicular tissue. This article reviews the classical and current terminology, philosophies, and techniques that are used at present to determine endodontic working length accurately, and describes their clinical application. Recent clinical advances such as the electronic apex locator, direct digital radiography, and the surgical operating microscope also are discussed. PMID- 10687441 TI - Synthetic bone grafts in peri-implant bone dehiscences: histological results in humans. AB - This study describes the histological results found in three patients treated with osseointegrated implants and Bioplant HTR (Hard Tissue Replacement) synthetic bone graft in peri-implant dehiscences adjacent to implants. This therapy was carried out without the use of barrier membranes. Bioplant HTR is reported to act as its own barrier and prevent gingival soft tissue migration ingrowth. The histologic picture demonstrated that Bioplant HTR is osteoconductive and biocompatible, and can be used both as bone substitute and as a barrier for guided bone regeneration in implant therapy. PMID- 10687442 TI - Anterior crossbite correction with fixed appliances in the adult dentition. AB - Anterior crossbite in the adult dentition, if not corrected, can cause other functional problems for the patient. These adult patients can be treated quickly and with confidence by most general dentists with the use of pre-adjusted appliances and straight wire. This article presents a clinical picture of three cases involving anterior crossbite correction in the adult patient with the use of fixed appliances and an acrylic splint. PMID- 10687443 TI - Microleakage of two new combined primer/adhesive resin systems. AB - In vitro microleakage of two new combined primer/adhesive systems utilizing the all etch technique (enamel and dentin etch) with the hybrid composite resin TPH is reported. These new combined primer/adhesive systems were compared to the Optibond FL system, which consists of a separate primer and adhesive. Forty Class V preparations were cut on the facial surfaces at the cemento-enamel junction of bovine incisor teeth to a dimension of 2.0 mm x 3.0 mm x 2.0 mm with a 1.0 mm 45 degree incisal bevel. The enamel (incisal) and dentin (gingival) margins were scored separately. Results suggest that Prime & Bond provided an improvement in the gingival marginal seal over the other materials. There was no statistically significant difference in the sealing ability of the materials tested at the incisal margin. PMID- 10687444 TI - A computerized model for teaching various methods of positioning the condyles to centric relation. AB - The entire masticatory system is within the realm of the dentist's responsibility, and therefore requires the dentist to have a working knowledge and understanding of the TMJs, neuromuscular system, and occlusion as well as the teeth and periodontal structures. This article presents how a computerized model can help dentists help their patients with TMJ disorders. PMID- 10687445 TI - Oral diagnosis you can't afford to miss. AB - The development of more predictable therapeutic modalities for the management of periodontitis has been preceded to a great extent by a better understanding of the underlying disease process. As the role of the immune mechanism has become more defined in the cascade of events associated with the progressing lesions, specific diagnostic tests are being developed to monitor the biochemical changes in the host. These tests, in addition to microbiological monitoring, together will help define therapeutic approaches, maintenance strategies, and endpoints in therapy. PMID- 10687446 TI - Atrial fibrillation: medications and dental considerations. PMID- 10687447 TI - Sexual boundaries in dental practice: Part 2. AB - Sexual harassment in the workplace is harmful to employees and disruptive. While the ethical obligations arising from the doctor-patient relationship do not address a dentist's ethical obligations toward his or her employees, the more general ethical obligations to the profession and those of business ethics would mediate against the creating or tolerance of sexual harassment in the workplace. The legal implications of participating in sexual harassment or allowing it to persist in the dental office are significant and dentists should be aware that employees enjoy legal protections against this inappropriate and potentially unlawful behavior. While any gender in any position may be the victim of sexual harassment, women who occupy subordinate positions are the most likely targets of these overtures. Dentist-employers are wise to be proactive in providing an appropriate protocol for employees to use when they perceive sexual harassment in the workplace. PMID- 10687448 TI - Seating, finishing, and polishing of ceramic restorations. AB - The insertion, finishing, and polishing of ceramic restorations can be arduous even for the most skilled operator. Attention to details as described above and refinement of all steps can lead to clinical success (Fig. 8-12). At this time composite and ceramic margins cannot equal those of cast or direct gold but with care and patience, clinically adequate results can be achieved. PMID- 10687449 TI - A comparison of all-ceramic restorative systems, Part 1. PMID- 10687450 TI - The oral implications of Hodgkin's disease. AB - Hodgkin's disease is a cancer involving the lymphatics. While the yearly total of new cases is only approximately 7,500, the disease is very curable with modern radiotherapy and/or combination chemotherapy. This represents an increasing number of survivors who require dental treatment. Unfortunately, the treatment for Hodgkin's results in significant, permanent complications that persist for the duration of the patient's life. These complications can include xerostomia, radiation-induced caries, the risk of osteoradionecrosis in irradiated bone, and systemic complications such as a reduced immune response to microorganisms. These patients can and do undergo all types of dental treatment safely as long as the practitioner recognizes the risks involved and takes appropriate precautions. PMID- 10687451 TI - Scleroderma: what the general dentist should know. AB - Scleroderma is characterized by fibrotic changes of the skin and organ systems. The disease presents in a variety of forms, ranging from the most aggressive progressive systemic sclerosis (PSS), to the three less aggressive, CREST syndrome, morphea, and the more localized form, linear scleroderma. The oral manifestations of the disease result from deposits of collagen in the tissues or as a result of collagen deposition around nerves and vessels. All oral tissues are affected. In scleroderma patients, the oral manifestation most commonly recognized is wide periodontal ligament spaces. Limited function, impaired healing, and neurologic symptoms all may be present to varying degrees, depending on the form and progression of the disease. PMID- 10687452 TI - Not all patients are the same: systemic risk factors for adult periodontitis. AB - Periodontal diseases are viewed today as multifactoral problems that are initiated and sustained by bacteria but significantly modified by the body's response to bacterial plaque. Local and systemic risk factors are involved in the disease process and both should be included when prognosis and treatment plans are developed. The most significant systemic modifying factors appear to be smoking, diabetes, and a recently discovered genetic marker. Slight changes in genes that produce an important inflammatory mediator, found in one-third of those studied, can lead to major negative outcomes in the way the body responds to bacteria. PMID- 10687453 TI - Nonsteroidal anti-inflammatory drug use in dentistry: gastrointestinal implications. AB - Pain management long has been an important consideration in dental care. Nonopioid analgesics such as aspirin, acetaminophen, and many of the nonsteroidal anti-inflammatory drugs (NSAIDs) generally provide predictable outcomes for control of dental pain because of their analgesic and anti-inflammatory properties. The anti-inflammatory action of NSAIDs also is being investigated in treatment of periodontal disease when used as an adjunct to non-surgical management. Since NSAID use by dental patients is high, gastrointestinal complications may arise, affecting long-term use of these agents. Systemic considerations of NSAID use are reviewed, as are complications frequently resulting in ulceration of the gastric mucosa. PMID- 10687454 TI - Dentists at high risk for hearing loss: protection with custom earplugs. AB - This article provides information on the effects of noise, especially to dentists. It gives information on how to protect one's hearing and furnishes a technique to fabricate custom earplugs. PMID- 10687455 TI - Simplifying the bead-brush technique. AB - This article describes a method of controlling monomer addition using the bead brush technique. With the addition of a cotton pellet to the monomer, incorporation of liquid can adequately be controlled. PMID- 10687456 TI - Resin-bonded fixed partial denture: a contemporary prosthesis. AB - This review of the evolution of the resin-bonded fixed partial denture (RBFPD) design, from the original "prepless" restoration to the present prosthesis, requiring careful treatment planning and technical skill, also discusses the materials used to enhance the longevity of this restoration. That, in turn, increases the patient's confidence in both the dentist and the prosthesis itself. PMID- 10687457 TI - Adult orthodontic treatment: a challenging case report. AB - The case of a 61-year-old woman with a primary diagnosis of spondyloepiphyseal dysplasia and severe maxillary and mandibular spacing as well as anterior and posterior crossbites is presented. Clinical findings led to a diagnosis of macroglossia which required partial glossectomy in addition to comprehensive orthodontic treatment to correct malocclusion. PMID- 10687458 TI - Enhanced fixed prosthetics with a connective tissue ridge augmentation. AB - Augmentation of the partially edentulous ridge can significantly improve the final prosthetic result. The subepithelial connective tissue graft is the treatment of choice to enhance soft tissue contours. The major advantage of the connective tissue graft is the preservation of the existing gingival coloration and tissue characteristics. A case is described that utilizes the subepithelial connective tissue graft for augmentation of a maxillary anterior ridge prior to prosthetic rehabilitation. PMID- 10687459 TI - In memoriam William W. Howard, DMD, MAGD. PMID- 10687460 TI - Antifungal drugs and fungal resistance: the need for a new generation of drugs. PMID- 10687461 TI - Ethical issues in the acceptance of gifts: Part 2. PMID- 10687462 TI - The use of indirect composite and ceramic inlays to restore anterior teeth: a clinical case report. AB - A clinical case has been presented utilizing old inlay techniques with new materials. The teeth were restored to proper form, function, and esthetics with a minimal removal of tooth structure. Crowns were not necessary, thereby eliminating a margin near the gingiva, and esthetics were acceptable. PMID- 10687463 TI - Evidence-based dentistry: an overview of a new approach to dental practice. AB - Dental students' clinical questions and problems are solved by a combination of instructors' intuition, training, and clinical experience, which may or may not be based on scientific evidence. This type of training, which relies heavily on clinical experience and information learned in dental school, seminars, or from colleagues, can lead to inappropriate treatment outcomes. Evidence-based dentistry attempts to answer clinical questions based on a critical review of the best available scientific evidence together with one's clinical experience and scientific knowledge. PMID- 10687464 TI - Adjunctive diagnostic methods for monitoring progressive periodontal diseases. AB - Diagnosing periodontal diseases involves determining the classification of the disease and the recognition of disease severity at the time of the clinical examination. Traditional diagnostic methods, such as radiographs and periodontal probing, have several limitations. Newer diagnostic techniques have been developed that could help determine whether specific sites are actively breaking down or if the site will experience future active disease. The newest technique is a genetic susceptibility test to identify patients who are at an increased risk for periodontal disease before problems develop. The screening of at-risk patients after initial therapy with these newly developed monitoring tests may lead to improved treatment and prevention of periodontal disease. PMID- 10687465 TI - Determining when to refer periodontal patients--clinical guidelines. AB - As part of the examination and evaluation of the patient, it is the dentist's responsibility to evaluate the periodontal tissues for the presence or absence of periodontal disease. If the practitioner finds that periodontal disease is present and is unwilling or unable to accurately diagnose and/or treat the type or extent of the disease, the dentist must offer the patient the opportunity to be evaluated by a practitioner who can complete the examination and diagnostic process. PMID- 10687466 TI - Interdisciplinary management of a common esthetic complaint. AB - Esthetic restoration of the maxillary anterior dentition often requires an integrated perio-restorative team approach. Careful attention to treatment planning and treatment sequencing is essential in ensuring a desirable outcome and a satisfied patient. Detailed measurements, diagnostic wax-ups, surgical mock ups, and stents all are effective communication tools which can be used to enhance interdisciplinary communication. These concepts and treatment modalities are illustrated in this case report. PMID- 10687467 TI - Repair of a root perforation with a resin-ionomer using an intentional replantation technique. AB - The repair of a root perforation can be accomplished using different materials and techniques. When the defect is surgically inaccessible, the tooth can be carefully extracted, repaired extraorally, and placed back into the socket. This procedure, known as intentional replantation, is often a measure of last resort in an heroic effort to save a hopeless tooth. This case report describes the treatment of a tooth with an iatrogenic root perforation and the subsequent healing of the surrounding periodontium using an intentional replantation technique and resin-ionomer to repair the root defect. PMID- 10687468 TI - Stable matrix for amalgam build-up. AB - A method is proposed of making amalgam build-ups for coronally debilitated teeth through the use of a stable matrix consisting of a disposable tray and polyvinylsiloxane occlusal registration material. For teeth with minimal remaining coronal tooth structure and/or the absence of adjacent teeth, this technique provides proper stabilization of the matrix band to permit adequate condensation of amalgam. PMID- 10687469 TI - Communicating with patients. AB - Dental care providers need to be able to communicate effectively with their patients in order to build rapport and trust. Highly developed communication skills also enable the dental care provider to extract more accurate diagnostic information and to more effectively present treatment options to the patient. Neurolinguistic programming techniques can be employed to accomplish these as well as other objectives. PMID- 10687470 TI - Fracture resistance of endodontically treated teeth restored with bonded amalgam and full crowns. AB - The use of dentin bonding prior to placement of core build-up restorations has been shown to reduce microleakage and reinforce remaining tooth structure. However, information is lacking about the influence a core build-up has on the fracture resistance of crowned teeth. The purpose of this in-vitro study was to compare the influence of three types of core build-ups on the fracture resistance of crowned teeth. PMID- 10687471 TI - Single appointment amalgam crown procedure for posterior teeth. AB - When faced with the restoration of severely broken-down posterior teeth, many treatment options exist. Expense, time constraints, and periodontal health, as well as the needs and desires of the patient, all play a role in the selection of treatment modality. In many instances, a well-fabricated amalgam crown can provide a highly acceptable functional restoration in the posterior region. Here, the amalgam crown is discussed in relation to other options and several case reports are presented. PMID- 10687472 TI - Oral Kaposi's sarcoma in a non-AIDS patient. AB - Kaposi's sarcoma involving the oral cavity is seen frequently in AIDS patients but rarely in transplant patients. When the oral cavity is involved in transplant patients, it usually is located on the palate or the oropharynx. This article reports a renal transplant patient who developed Kaposi's sarcoma which mimicked a gingival hyperplasia in the oral cavity. PMID- 10687473 TI - Tobacco "plantibodies" for caries prevention. PMID- 10687475 TI - Effects of light intensity, time, and direction on gap formation of resin composite restorations. PMID- 10687474 TI - Sexual boundaries in dental practice: Part 1. AB - Sexual contact between doctors and patients is unethical, legally perilous, cause for professional discipline at times, and often viewed as an outrageous transgression by the public. While it is true that some provider-patient couples go on to get married and "live happily ever after," this is not always the case. However, if these relationships fail, a now embittered ex-paramour is empowered with the options of bringing legal and/or disciplinary action against the doctor and may be motivated by the distinct possibility of significant financial reward. Patients place enormous trust and respect in their health care providers. In addition, patients reveal sensitive, confidential information to doctors and do so with the expectation that it will be used only for their best interests. This dynamic creates a substantial power imbalance between doctors and patients and this power differential must never be exploited. Dentists who find themselves romantically attracted to patients must either avoid initiating a more intimate relationship or refer the patient to another provider. Dentists who are the recipients of romantic inquiries by patients should establish clear boundaries. If the dentist is available and interested in the patient, dating may occur only after the patient has been reassigned to another dentist and a suitable time period has elapsed. PMID- 10687476 TI - Treatment plan case report. PMID- 10687477 TI - A day at the office (an advertorial). AB - This issue of General Dentistry highlights how tobacco products affect oral health. The following articles are presented to help general dentists with their efforts to champion cessation and to educate their patients. Please also note the guest editorial from Dr. Robert Mecklenburg. This first article by Dr. Stephen B. Corbin is an introduction to the topic at large and provides the reader with a suggested script for use in helping patients quit the smoking habit. We hope you find these articles useful and look forward to hearing your response. Please fax (312/440-4261) or e-mail (AGDJournal@agd.org) your thoughts on these articles. We hope these articles succeed in helping you help your patients beat the tobacco habit. PMID- 10687478 TI - Tobacco cessation: a practical dental service. AB - Tobacco use is a complex addiction that must be addressed in all aspects of health care. Despite the deleterious and costly outcomes of tobacco use, Americans still are smoking and using smokeless tobacco. Dentists are trained to detect oral lesions and periodontal problems that are related to tobacco use. Dentists also are in a position to help prevent the initiation of tobacco use by children and adolescents through the use of positive anti-tobacco messages. Over the past decade, tobacco cessation strategies have been modified for practical use in the dental setting. PMID- 10687479 TI - Tobacco and dental implants. AB - Dental implants are the ideal standard of care for many oral health care providers. Tobacco use is an impediment to the success of this sophisticated procedure. Dentists who are trained to help their patients stop using tobacco are in position to improve their success rates with dental implants. A suggested protocol for tobacco cessation in the implant practice, if utilized, could raise the standard of health care in the dental office. PMID- 10687480 TI - Canal configuration of the mesiobuccal root of the maxillary first molar: conventional and surgical approaches. AB - One of the primary causes of endodontic failure is the presence of untreated canals. Although the maxillary first molar has been described as the most endodontically treated tooth, the canal configuration has been the least understood. The purpose of this article is to present, with the aid of several clinical cases, a review of the literature regarding the frequency, the location, and the incidence of the second mesiobuccal canal in relation to the maxillary first molar, both for conventional and surgical approach. PMID- 10687481 TI - Prevalence and implications of accessory retromolar foramina in clinical dentistry. AB - The occurrence of retromolar foramina (RMF) was examined in a sample of dry skulls (African American n = 249; Causcasian n = 226) to consider the potential clinical impact. A prevalence rate of 7.8% of RMF was found. There were no statistical differences based on race or gender. The prevalence may contribute to the explanation of a portion of inferior alveolar nerve block failures and provide insight into potential implications of surgery in the posterior mandible. PMID- 10687482 TI - Using soft splints in your dental practice. AB - A significant number of soft splints are fabricated by U.S. dentists every year. The efficacy of these splints is discussed, reported indications and contraindications are defined, and favorable and unfavorable characteristics reported by patients are presented. Maxillary and mandibular soft splint designs that have been found to be clinically acceptable are illustrated and an easy and rapid technique for adjusting and polishing a soft splint is proposed. PMID- 10687483 TI - The free gingival graft combined with the frenectomy: a clinical review. AB - Abnormal labial frena are capable of retracting gingival margins, creating diastemas, and limiting lip movement. When these frena are present, the traditional frenectomy alone generally is successful. However, when the frenulum is extensive, the possibility of coronal reformation exists. Several procedures have combined the frenectomy with either a lateral pedicle flap, free papilla graft, or free gingival (mucosal) graft taken from the palate. Three case reports demonstrate the continued efficacy of the traditional palatal free gingival graft when the patient has an extensive frenulum or an area of minimal esthetic concern is involved. PMID- 10687484 TI - Effective use of the ortho-panoramic radiograph during the doctor-patient conference. AB - Tracings of pre-treatment radiographs can be used to clarify the case presentation and be inserted into the permanent record. The use of different colors on the photocopy can highlight or demonstrate various conditions or steps in treatment to enhance the patient's understanding. Various other uses for such tracings are suggested. PMID- 10687485 TI - Everything you wanted to know about radiographic duplication. AB - If dental radiographs are not duplicated correctly, the resulting duplicates will be of inferior quality. This article discusses the characteristics of a duplicating film, the principle of radiographic duplication, a description of dental duplicators, the selection of optimum duplicating time, a technique of duplicating radiographs, an alternate technique of duplicating radiographs, errors in duplication and their correction, and the maintenance of duplicators. PMID- 10687486 TI - Gingival augmentation with a dermal allograft. AB - Gingival augmentation surgery with soft tissue autografts long has been the standard for increasing the width of keratinized oral tissue. Acellular dermal allografts, which have been used for several years by reconstructive surgeons, are a novel technique for achieving increased gingival tissue in place of soft tissue autografts. This report describes a case of gingival augmentation via an acellular dermal allograft. Reduced morbidity from donor site grafts and increased patient acceptance, along with highly successful clinical results, are the primary advantages of this acellular dermal allograft. PMID- 10687487 TI - Recent reports calling for reduction of antibiotic prophylaxis in dental procedures: a response. PMID- 10687488 TI - Managed care--serving two masters. AB - It is likely that enrollment in managed care dental plans will continue to increase. Dentistry can respond to this trend by resisting it; however, just as the health care marketplace drove medical care into managed care mechanisms, so too will these forces impact dentistry. For those who are participating in managed care dental plans, it is heartening that current data indicate that most types of patient care are not adversely affected by reimbursement mechanisms. Dentistry, however, should seek out opportunities to shape the managed care format and must be at the table to assure that ethical principles and conflicts of interest receive due consideration. Dentists who treat patients under a managed care reimbursement system must be certain that the plan does not require providers to sacrifice patient autonomy or compromise care in the process of serving two masters. PMID- 10687489 TI - Indirect provisional restorations. AB - Many methods of provisional restoration fabrication are available to practicing dentists. The indirect method has been found to be the most accurate and least time consuming for the author as well as many other clinicians. After a little practice and familiarity with the methods and materials the restorative dentist will realize the benefits for both the dentist and the patient. PMID- 10687490 TI - Percutaneous injuries among dental health care workers. AB - To evaluate percutaneous injuries among dental health care workers this survey posed two questions: Is there a difference in the number of percutaneous injuries occurring among dentists, dental hygienists, and dental assistants, and Is there a difference in the number of injuries that occur intraorally or extraorally among dental health care workers as a whole, and within each occupational group? This prospective study included demographic information and an incident report. The incident report tabulated type of injury (intraoral or extraoral), procedure during which the injury occurred, and type of instrument that caused the injury. PMID- 10687491 TI - Disinfection and monitoring of dental unit waterlines. AB - Recent increased awareness of dental unit waterline contamination has prompted investigations into potential protocols for improvement in the quality of water delivered to patients. When the protocol was followed (n = 12), contamination levels were reduced to below measurable colony forming units (CFU) defined as < 1 CFU/mL within three weeks and maintained thereafter. When adherence to the protocol was not strictly followed after initial disinfection, contamination quickly returned and remained until additional disinfection was performed. PMID- 10687492 TI - Evaluation of sterilization of dental handpieces by heating in synthetic compressor lubricant. AB - The Centers for Disease Control and Prevention and the American Dental Association guidelines recommend sterilization of dental handpieces after each use. Steam autoclaving is the most commonly used sterilization method. However, pressurized steam causes corrosion and partial combustion of the handpiece lubricant, leaving a sticky carbon residue on the turbine which must then be replaced after several usages. Replacement of autoclave-damaged dental handpieces represents a major expense for dentists that may be avoided through the use of less destructive sterilization techniques. PMID- 10687493 TI - Irrigation with antimicrobial agents for the treatment of periodontitis--is it effective? AB - Subgingival irrigation has been proposed as a beneficial adjunct to scaling and root planing or ultrasonic scaling. The most commonly investigated agents are iodine and chlorhexidine. Clinical studies from the past 15 years are reviewed to determine the real benefits of antimicrobial irrigants in conjunction with root planning. With knowledge of the treatment protocols and results of these clinical studies, the clinician is equipped with a biologic rationale for his treatment decisions in nonsurgical periodontics. PMID- 10687494 TI - Site specific delivery of antimicrobial agents for periodontal disease. AB - Controlled release delivery systems have become available for the sustained delivery of antimicrobial agents directly to the periodontal pocket. These systems have shown clinical efficacy in periodontal therapy both as adjunctive treatments and as stand-alone therapies. A review of the current state-of-the-art of site specific delivery of antimicrobial agents for the treatment of periodontal disease is provided. PMID- 10687495 TI - Differential diagnosis of pericoronal radiolucencies with and without radiopacities. AB - Pericoronal radiolucencies, by definition, surround the crown of a tooth. They are encountered not only in pediatric patients but also in adults of all ages. It is likely that all clinicians, at some time or another, will be confronted with a radiograph showing a pericoronal radiolucent lesion. The differential diagnosis of such lesions is reviewed. PMID- 10687496 TI - Antimicrobial activity of cavity disinfectants. AB - The purpose of this study was to determine the antimicrobial activity of four commercially available cavity disinfectants and one prescription mouthwash as they came into contact with bacteria commonly found in the oral cavity. Streptococcus mutans, salivarius, and Actinomyces viscosus were used in the study. Zones of microbial inhibition were measured in millimeters after 48 hours. The results of this study indicate that all of the antimicrobial agents demonstrated activity against the bacteria tested. Consepsis Solution produced the largest zones of inhibition against all three of the bacteria used. PMID- 10687497 TI - To place provisionals or not--that is the question. AB - Porcelain labial veneers are quite possibly the most esthetic restorations that can be provided today. Their esthetic qualities are derived not only from the accurate replication of tooth structure, but also from the superior tissue response they elicit. Many of the procedures surrounding this restoration have become almost standardized in terms of preparation and the manner in which the restoration is made to become part of the tooth. But why experts who lecture about this procedure remain at odds about whether provisional restorations are necessary is baffling. As more tooth preparation became increasingly necessary to achieve the ultimate in esthetics and health, use of the provisional restoration was not incorporated, simply as a matter of established routine. PMID- 10687498 TI - Effect of glutaraldehyde-based cold sterilization solutions on light transmission of single-use, plastic light-curing tips. AB - Previous research has demonstrated that autoclaving of conventional curing tips results in the build-up of an opaque scale on the curing tips ends, greatly reducing light intensity output. Cold sterilization of conventional light-curing tips in most glutaraldehyde-based solutions did not damage the tip or decrease light-transmission quality. However, the specific brand of cold sterilant was found to be of importance, as one specific product was shown to decrease light tip intensity values irreversibly, even after subsequent tip polishing. A decrease in light intensity output from the curing source significantly affects polymer cure and the biological properties of the restorative material. PMID- 10687499 TI - Treating enamel surfaces with a prepared pumice prophy paste prior to bonding. AB - The use of pumice to remove the salivary pellicle, plaque, and/or surface debris is a well-known procedure. However, pumice can act as a contaminant. Therefore, a slurry of pumice without additives is recommended for use prior to bonding procedures. This article presents a review of the shear bond strengths obtained by bonding composite resin to enamel after the enamel had been cleansed with a slurry of pumice and a premixed caplet of pumice. PMID- 10687500 TI - A new and indirect working die technique for fabricating temporary restorations. AB - A time- and labor-saving indirect method for fabricating temporary restorations that features removable dies (coined the indirect working die technique), is introduced and described. It takes advantage of the accuracy of the traditional indirect method, as substantiated in the literature, but employs new methods and uses materials in a novel way. PMID- 10687501 TI - Dual-purpose, radiographic-surgical implant template: fabrication technique. AB - A technique for fabrication of an implant radiographic-surgical template for the partially-edentulous patient is proposed. A mixture of barium sulfate and acrylic resin provides a radiopaque template of the planned implant restorations in relation to the hard tissues. This template is easily converted to a surgical guide. PMID- 10687502 TI - The impact of managed care in dentistry. AB - Managed care plans attempt to control health care expenditures aggressively. These plans directly influence access to medical care and the type, level, and frequency of care rendered. As a result, hospital stays are reduced, focus shifts from inpatient to outpatient care, and patients are responsible for a larger share of health care costs. Dentistry is not immune from the impact of managed care. The attractiveness of the dental market has drawn many managed care organizations, insurers, and entrepreneurs to encourage dentists to participate in a wide variety of managed care programs. However, the delivery of dental care differs markedly in many respects from that of medical care. Therefore, many of the cost saving aspects of managed care that have been so successful in medicine may not result in similar cost savings in dentistry. PMID- 10687503 TI - Double blind clinical trial of a remineralizing dentifrice in the prevention of caries in a radiation therapy population. AB - OBJECTIVES: The purpose of this study is to determine the efficacy and safety of a specially formulated remineralizing toothpaste in controlling caries in a group of high risk, head and neck radiation patients. DESIGN: The study compares the performance of the remineralizing toothpaste with a leading conventional fluoride dentifrice using double-blind randomization. TEST PRODUCTS: The products compared both contain equivalent quantities of fluoride (1150 ppm). The remineralizing toothpaste also delivers soluble calcium and phosphate ions, the essential components of teeth. SUBJECTS: On completion, 50 subjects who received > 50 Gy of radiation to the head and neck. MEASUREMENTS: Examinations include coronal and root caries using the Pitts Diagnostic Criteria, salivary flow rate, plaque and gingival indices and microbiological counts over one year. RESULTS: At this point subjects are enrolled in the study at various phases. However, the current average for the net increment per month per subject is -0.12 (+/- 1.30) for coronal caries and 0.06 (+/- 0.73) for root caries in subjects using the remineralizing toothpaste and 0.53 (+/- 1.62) for coronal caries and 0.45 (+/- 0.98) for root caries in subjects using the conventional fluoride dentifrice. Non parametric analysis of rank scores for net root surface increments/month was statistically significant (p = 0.02), suggesting lower net root surface increment/month for the remineralizing toothpaste relative to the conventional toothpaste. No significant differences were noted on coronal surfaces. CONCLUSIONS: The results to date indicate that the remineralizing toothpaste is significantly superior to the conventional fluoride dentifrice in preventing root caries in high risk patients. PMID- 10687504 TI - The impact of oral health on stated ability to eat certain foods; findings from the National Diet and Nutrition Survey of Older People in Great Britain. AB - OBJECTIVES: To assess how the dental status of older people's mouths affected their stated ability to eat common foods. DESIGN: Cross sectional study. SUBJECTS: Survey was part of the oral health component of the nationwide British National Diet and Nutrition Survey: people aged 65 years and older. Two separate representative samples aged 65 and over: a free-living and an institutional sample. 881 free-living and 275 institution subjects had a dental exam and were interviewed about ability to eat key foods. RESULTS: Significant percentages of free-living people had difficulty or could not eat at least 4 of 16 foods, and about 1 in 5 dentate stated they had difficulty eating or could not eat raw carrots, apples, well-done steak or nuts. More of the edentate subjects stated that they had difficulty eating than the dentate. Perceived chewing ability increased with increasing numbers of natural teeth and pairs of opposing posterior teeth. Subjects reporting a sociodental impact were more likely to consider that they were unable to eat foods that required more chewing. Associations remained valid after correction for the effects of age, sex, social class and denture wearing status and region. Perceived dryness did not affect significantly the stated ease of eating most foods. There were more dietary restrictions reported by the institution sample. Some foods, such as nuts, apples and raw carrots could not be eaten easily by over half of edentate people in the institution sample. CONCLUSIONS: The stated selection of foods are substantially affected by numbers of teeth and occluding pairs of posterior teeth and presence of full dentures in significant percentages of older people. PMID- 10687505 TI - Socio-medical condition and oral functional status in an older institutionalised population. AB - OBJECTIVES: To investigate the association between the general medical condition, the socio-economic status, and some factors related to the functional status of the stomatognathic system. DESIGN: A survey in an elderly population. SUBJECTS: 257 older adults, with a mean age of 83.7 years. SETTING: Residential homes for the elderly. INTERVENTION: Examination of the medical records on the overall health and the drugs consumed, a structured interview on the socio-economic status, the complaints for xerostomia, the subjective chewing difficulties, and a clinical evaluation of the number of natural teeth and the number of posterior occluding pairs of teeth contacts (premolars and molars). RESULTS: Multiple pathology and polypharmacy were recorded. 25% of the residents had no occluding posterior tooth contact (natural or prosthetic) and 62% were edentulous. 43% of the residents reported complaints for xerostomia, and 46% for chewing difficulties when eating specific food types. Xerostomic feelings and chewing problems were not related to age. Chewing difficulties were not related to the number of natural teeth, but to the number of posterior occluding teeth contacts, natural or prosthetic (less than two). From all medical conditions examined, only the psychiatric disorders were significantly related to dental status (p < 0.05). Moreover, the number of remaining natural teeth was related to socio-economic status, while the number of posterior occluding teeth contacts was also inversely related to the duration of institutionalisation (p < 0.05). CONCLUSIONS: Psychiatric disorders, low socio-economic status and increased duration of institutionalisation were most closely related to poor dental status. The presence of more than two posterior occluding teeth contacts, natural or prosthetic, benefit the very old patient in terms of reduced subjective chewing difficulty. A formal oral care delivery system for the institutionalised elderly, and particularly for those suffering from psychiatric disorders, is imperative. PMID- 10687506 TI - Age-related changes in cellular activity in human submandibular glands as evaluated by argyrophilic nucleolar organizer regions. AB - OBJECTIVE: To examine the age-related changes in cellular activity of epithelial components of human submandibular glands, evaluated on the basis of argyrophilic nucleolar organizer regions (AgNORs). DESIGN: Epithelial components of human submandibular glands were divided into serous acinar cells, mucous acinar cells, intercalated duct cells, striated duct cells, and interlobular duct cells. The mean AgNOR number of each cell type was compared among six age groups. SETTING: The study was conducted at the Department of Oral Pathology, Tohoku University School of Dentistry, Japan. SUBJECTS: Necropsy specimens from 66 males and 57 females 1 to 97 years old. RESULTS: In all cell types except for intercalated duct cells, the mean AgNOR number was lowest in the 0-14 year-old group and highest in the 15-29 year-old group. The value then gradually decreased with advancing age and ultimately reached a similar level to that in the 0-14 year-old group. In intercalated duct cells, the mean AgNOR number did not differ significantly between any age group. There were no significant sex-related differences. CONCLUSIONS: The cellular activity of almost all components of human submandibular glands rises in adolescence and young adulthood and then decreases with aging. These results suggest that intercalated duct cells are capable of not only proliferation but also division into other components; these cells may thus compensate for the reduced activity of other components in elderly subjects. PMID- 10687507 TI - Factors which are associated with dental decay in the older individual. AB - OBJECTIVES: To improve reliability of salivary bacterial cultures as a surrogate for plaque levels of cariogenic bacterial species by reporting the salivary CFUs of these organisms as a function of the number of teeth. DESIGN: Cross-sectional collection of data in a convenience sample of adults over 60 years of age. SETTING: Hospital Dental clinic, University bacteriology laboratory. SUBJECTS: 523 older dentate subjects, average age 70, including 412 subjects who were in an independent living status and 111 in a dependent-living situation. MAIN OUTCOME MEASURES: Subjects were examined for decay and the presence of salivary factors including the levels of S. mutans, lactobacilli, yeast and other bacteria. The salivary levels of the bacteria were adjusted for the number of teeth in the mouth, and the resultant values were entered into multivariable logistic regression models along with clinical and other salivary parameters. RESULTS: Mutans streptococci levels reported as CFUs/ml saliva per tooth were significantly associated with coronal surface decay, and lactobacilli, reported in a similar way, were significantly associated with root surface decay. Salivary levels of yeasts, which had previously been associated with decay in this population, were no longer significant using this construct. CONCLUSIONS: This construct of reporting salivary bacteriological data as a function of tooth number and per ml saliva could improve the reliability of bacteriological data obtained in epidemiological studies investigating the role of bacteria in dental decay in the elderly. PMID- 10687508 TI - Caries activity and associated risk factors in elderly hospitalised population- 15-months follow-up in French institutions. AB - Only a few studies have been published concerning hospitalised elderly disabled people. OBJECTIVES: 1) to investigate the oral health status of elderly French patients hospitalised in the two main geriatric hospitals of Paris. 2) to describe the respective influences of general parameters (type of hospitalisation, pathologies and medication) on oral environment parameters. 3) to analyse the influences of these oral parameters on caries activity in Long Term Care (LTCF) and in rehabilitation facilities (RF) patients and to study the incidence and the time-course of caries in these specific population. SUBJECTS: 117 subjects (mean age = 83.0 years, SD = 7.8, range = 64 to 102 years) were examined at baseline and 32 of the 50 LTCF subjects were reexamined 15-months later. METHODS: The general parameters recorded were age, gender, type of hospitalisation, period of stay, removable prosthesis, general diseases, number of diagnoses, medications with hyposalivary side-effects. The oral environment parameters recorded were flow rate, buffer capacity, mutans streptococci and lactobacilli counts, measured at baseline by tests on stimulated saliva, and plaque index. Crown and root surfaces were recorded according to a modified caries activity index. RESULTS: Among the polypathological subjects (85.5% of the population), the number of diseases ranged from 2 to 8. The LTCF patients had a significantly higher mean number of diagnoses (3.5; SD = 1.5) than the RF patients (2.8; SD = 1.4). 76.9% of patients were taking medications with hyposalivary side-effects. The stimulated flow rate ranged from 0.02 ml/min to 5 ml/min. Its mean was significantly lower for LTCF patients (0.67 ml/min; SD = 0.51) than for RF patients (1.12 ml/min; SD = 0.89). The plaque index was significantly higher in LTCF subjects and in patients with mental diseases. At baseline, 17,442 crown and root surfaces were examined. Flow rate was related to crown caries and buffer capacity to root caries. During the 15-months follow-up, the mean number of active root surfaces was significantly increased: from 0.148 (SD = 0.116) at baseline vs. 0.250 (SD = 0.174) at the second examination. CONCLUSIONS: The strongest relationship in the present study between oral parameters and caries activity was the negative relationship between buffer capacity and active root caries. This study confirms an association between the type of hospitalisation and both salivary parameters flow rate and plaque index. This investigation illustrates the critical need for hygiene and oral care, in this elderly disabled population. PMID- 10687509 TI - The importance of oral health to older people's quality of life. AB - OBJECTIVES: This study was designed to determine whether older people perceive oral health as being important to Quality of Life (QoL) and if so, to identify the most important ways in which their lives are affected. In addition, to identify if subgroups of older people perceive its importance differently. DESIGN: Nationwide qualitative face to face interviews with older people were carried out utilising the Office for National Statistics Omnibus survey in Great Britain. SUBJECTS AND METHODS: 454 adults aged 65 or older took part in this study, part of a random probability sample of adults in the UK. SETTING: Respondents were interviewed in their homes. RESULTS: 72% (313) perceived their oral health status as important to their QoL through a variety of physical, social and psychological ways. Most frequently its impact on function: eating (29%, 126) and symptoms: comfort (14%, 59) were considered most important. Gender and social class variations were apparent (P < 0.05). CONCLUSION: Older people perceive oral health as being important to life quality in a variety of different ways. There are significant social class and gender variations which must be taken into consideration when assessing oral health needs of older people. PMID- 10687510 TI - Effect of different grades of gum rosins and hydrogenated resins on the solubility, disintegration, and dimensional alterations of Grossman cement. AB - In the present study, we investigated the effect of the addition of different grades of gum rosins and hydrogenated resins to Grossman cement on dimensional stability, solubility and disintegration. pH and conductivity, which may affect these properties, were also determined. The experiments were performed according to Specification 57 of the American Dental Association for root canal cements using Grossman cements containing three gum rosins (grades X, WW, and WG) and two hydrogenated resins (Staybelite and Staybelite ester 10). The results showed that the solubility, disintegration, and dimensional stability of Grossman cement containing Staybelite and Staybelite ester 10 were inferior to the values considered acceptable by the American Dental Association Specification 57. PMID- 10687511 TI - In vitro release of hydroxyl ions from calcium hydroxide gutta-percha points. AB - In endodontic practice, calcium hydroxide is widely used for a number of reasons associated with its high pH. The purpose of the present study was to determine in vitro the alkalizing potential of newly introduced calcium hydroxide gutta-percha points that are proposed for temporary filling of root canals. The materials tested were: calcium hydroxide gutta-percha points; chemical pure calcium hydroxide powder mixed with distilled water; and Reogan rapid, a nonsetting calcium hydroxide preparation. The materials were placed into dialysis tubing and transferred into plastic vials containing bidistilled water. Measurements were taken by a digital pH meter after 10, 20, and 30 s; 1, 15, and 30 min; and 1, 2, 3, 24, 48, 72, 96, and 120 h. The calcium hydroxide containing gutta-percha points showed a significantly lower alkalizing potential than Reogan rapid and calcium hydroxide mixed with distilled water (p < 0.05). PMID- 10687512 TI - Scattering of laser light directed onto the labial surface of extracted human upper central incisors. AB - The purpose of this study was to evaluate the scatter of light through human teeth with laser Doppler flowmetry. An optical probe (probe 1), which mounted two optical fibers (one for light illumination and another for the measurement of backscattered light intensity) was placed on the labial surface of the extracted teeth. Another optical probe (probe 2) for the measurement of transmitted-light intensity was placed either on the mesial, distal, or palatal surface or in the canal of the teeth. The light intensity at probe 1 was stable, irrespective of the location of probe 2, whereas the transmitted light intensity at probe 2 tended to increase as probe 2 moved to more incisal positions on the mesial, distal, and palatal tooth surfaces. The results indicate that the light scatters to a wide area outside the tooth and provides information regarding the surrounding tissue blood flow. PMID- 10687513 TI - Changes in the periodontal membrane due to apical periodontitis. AB - Teeth with an apical inflammatory lesion were studied by light microscopic morphometrical procedures to estimate the volumetric density of periodontal ligament tissues by point counting. Sixty-four root surfaces were investigated from coronal to apical. The observed tissue changes were similar in groups with and without bacteria, except for an elevated volumetric density of inflammatory cells in the first group. The attachment was lost apically. Principal fibers running to the root surface and extensions in the cementum decreased from coronal to apical, but were replaced by fibers running parallel to cementum and fibers oriented in a network. We suggest that acellular extrinsic fiber cementum was lost, whereas cellular mixed stratified cementum was built. The cellular mixed stratified cementum synthesis was by inclusion of the remaining fibers from the acellular extrinsic fiber cementum. We suspect that the changes were the result of the anti-inflammatory reaction caused by the periapical lesion. PMID- 10687514 TI - Bactericidal effects of 2.94 microns Er:YAG-laser radiation in dental root canals. AB - The purpose of this study was to investigate the antimicrobial properties of Er:YAG-laser radiation in dental root canals. The root canals of 90 freshly extracted anterior teeth were enlarged mechanically, sterilized, and randomly divided into subgroups of 10 samples. The root canals were inoculated with Escherichia coli or Staphylococcus aureus for 2 h. The laser treatment groups were exposed for either 15 or 60 s to Er:YAG-laser radiation (pulse energy: 50 mJ; 15 pps). Additionally, for each bacterial strain, one sample group was rinsed with a NaOCl solution (1.25%), and one was left untreated as control. After irradiation or irrigation, the number of bacteria was evaluated using the surface spread plate technique. In the case of S. aureus, the primary bacterial load (control group) of the root canals was reduced to 0.15% after 15 s and 0.06% after 60 s of laser treatment. In the E. coli group, the number of bacteria was diminished to 0.13%, with the shorter radiation time and to 0.034% after 60 s of radiation. Irrigating the root canals with NaOCl, a reduction of the number of bacteria to 0.033% for S. aureus and to 0.020% for E. coli could be obtained. As the results confirm, Er:YAG-laser radiation exerts very effective antimicrobial properties in dental root canals, depending on the time of radiation. PMID- 10687515 TI - Wear of nickel-titanium lightspeed instruments evaluated by scanning electron microscopy. AB - Used rotary nickel-titanium instruments require frequent replacing. This laboratory study evaluated defects of Lightspeed cutting tips before and after usage. The instruments were fixed into custom-made holders, the cutting heads photographed in a scanning electron microscope at x120 to x400 magnification at preset points around the cutting tip (90, 180, 270 and 360 degrees) and head-on. Instrument sizes 20 to 32.5, 35 to 60, and 65 to 100 were used in 9, 18, and 36 canals, respectively, and autoclaved after shaping every third root canal. The used instruments were cleaned and then reexamined in a scanning electron microscope as before. The presence of 11 types of conditions was scored from the pre- and postusage photographs. No instruments fractured during the test, but all the cutting heads had one or more imperfections, even before usage. The presence of debris, pitting, and metal strips changed significantly. Imperfections were found on new and used Lightspeed cutting heads, indicating the general difficulty in machining defect-free nickel-titanium rotary instruments. However, high quality should remain a goal to improve instrument efficiency. PMID- 10687516 TI - Rapid decontamination of gutta-percha cones with sodium hypochlorite. AB - Gutta-percha cones are now widely used to fill root canals. Because they cannot be sterilized by conventional autoclaving or in a hot-air oven, gutta-percha cones require rapid chairside decontamination before use to maintain the aseptic chain, an essential factor in successful endodontic therapy. The purpose of this study was to compare the effectiveness of different concentrations of sodium hypochlorite (0.25% to 4%) in sterilizing gutta-percha cones artificially contaminated with Staphylococcus aureus and Escherichia coli strains, and Bacillus subtilis spores. After 1 min of treatment, the solutions tested showed bactericidal and sporicidal effects at concentrations of 0.25% and 1%, respectively. At a concentration of 0.25%, the solutions tested were effective in destroying spores after 5 min of exposure. Based on this study, treatment of the cones for 1 min with 1% sodium hypochlorite (Milton's solution) or for 5 min with Dakin's liquid (0.5% sodium hypochlorite) is recommended. PMID- 10687517 TI - Residual thickness of root in first maxillary premolars with post space preparation. AB - It has been suggested that, to ensure tooth strength, a minimum of 1 mm of root wall thickness should be left after post preparations. The purpose of this study was to determine the instrument diameter that will not affect this measurement in maxillary first premolars. Post preparations were made in 106 teeth with one and two root canals at a working depth equal to the anatomical crown length, with 0.70, 0.90, 1.10, 1.30, 1.50, and 1.70 mm diameter instruments. Sections were cut perpendicular to the long axis of the tooth at the cervical and apical ends of each preparation, and the minimum width of residual root was measured on each wall, at both sites. A binocular microscope with a micrometer eyepiece was used. At the cervical level of the preparation, no group showed a wall thickness < 1 mm. Data for the apical sections was statistically analyzed, and the corresponding confidence limits were calculated with 95% confidence on the mean. The results show that the minimum residual thickness was only preserved when 0.70 mm instruments were used in single-canal roots and when 1.10 mm or smaller instruments were used for two-canal roots. This seemingly anomalous result occurs because fluting on both the mesial and distal sides of the root impinge on single canals, whereas dual canals are buccally or lingually displaced to an area of thicker root diameter. PMID- 10687518 TI - An evaluation of endodontically treated vertically fractured teeth. AB - For this survey, 92 vertically fractured endodontically treated teeth were evaluated clinically and radiographically before and after extraction. The maxillary second premolars (27.2%) and mesial roots of the mandibular molars (24%) were the most fractured teeth. In 67.4% of the teeth, a solitary buccal pocket was present; in 34.8%, a fistula frequently appeared closer to the gingival margin than to the apical area. A lateral radiolucency or a combination of lateral and periapical radiolucency was found in more than half of the cases. The general practitioners correctly diagnosed vertical root fracture in only one third of the 92 fractured teeth in this survey. PMID- 10687519 TI - Surgical extrusion of a cervically root-fractured tooth after apexification treatment. AB - A case is reported in which an incisor fractured below the alveolar crest 6 months after completion of apexification treatment was surgically extruded for prosthetic coronal restoration. After the surgical procedure, a dowel post was placed in the root canal, a core was built using glass-ionomer cement, and a porcelain veneer crown restoration was completed. The 24-month follow-up examination after surgical, endodontic, and prosthetic treatments showed that the tooth was clinically and radiographically healthy and functioned well. PMID- 10687520 TI - Histological and densitometric analysis of the effects of exogeneous heparin on rat pulp tissues. AB - In this study, the effects of heparin on dentinal resorption was investigated in rats. Animals were injected with 2 x 2 IU/g Na-Heparin subcutaneously for 33 days. Histopathological examination of the anterior teeth revealed capillary proliferation, congestion, and fibrosis in the pulp in addition to resorptive lacunae and degenerative bone spiculae in the peripheral bone tissue. Development of fibrosis in the pulp tissue was verified by densitometric analysis, which revealed a 30% increases in mean density after heparin administration. No resorption at all, however, was seen in dentin. PMID- 10687521 TI - An evaluation of antimicrobial efficiency of Endo-Fill root canal sealant and filling material. AB - The antimicrobial efficiency of Endo-Fill root canal sealant and filling material was microbiologically evaluated. The zones of inhibition around the Endo-Fill by agar diffusion method were measured. Staphylococcus aureus, Streptococcus faecalis, Streptococcus pyogenes, Escherichia coli, Candiada albicans, and Pseudomonas aeruginosa were used as the selected microorganisms. No zone of inhibition was seen around the Endo-Fill in any of the examples. PMID- 10687522 TI - Bacterial microleakage of Cavit, IRM, TERM, and Fermit: a 21-day in vitro study. AB - The aim of our study was to evaluate the leakage of four cements (Cavit, IRM, TERM, and Fermit) using a two-compartment model system and Streptococcus sanguis as bacterial marker. Access cavities in premolars were filled with cement and the teeth immersed in culture medium in the model system. Half of the teeth were thermocycled on day 2. Bacterial percolation into the upper compartment was measured at regular intervals (days 2, 7, 14, and 21). Cement thickness was measured at the end of the study. In the nonthermocycled group, Cavit was more leakproof than the other cements at day 2 (p = 0.011), than TERM and IRM at day 7 (p = 0.043). Fermit was more leakproof than IRM at day 7 (p = 0.043). In the thermocycled group, Cavit was more leakproof than the other cements at day 7 (p = 0.041). Thermocycling did not significantly affect leakage. Cement thickness averaged 4.1 mm and did not significantly affect leakage. These results should be considered when using cements as temporary fillings. PMID- 10687523 TI - Evaluation of the combination of flurbiprofen and tramadol for management of endodontic pain. AB - Effective management of endodontic pain represents a continuing challenge. In this study, we evaluated the efficacy of flurbiprofen and a novel centrally acting analgesic, tramadol, alone and in combination, for reducing pain in endodontic emergency patients. Patients (n = 49) were administered a local anesthetic and underwent pulpectomy. They were then administered, on a double blind basis, either: (i) placebo (one capsule to start and then every 6 h); (ii) flurbiprofen (100 mg loading dose and then 50 mg every 6 h); (iii) tramadol (100 mg loading dose and then 100 mg every 6 h); or (iv) the combination of flurbiprofen and tramadol (as above). Pulpectomy combined with placebo medication resulted in a 50% reduction in pain by 24 h (p < 0.01). Patients treated with flurbiprofen and tramadol reported less pain, compared with placebo treatment at 6 and 24 h (p < 0.01 for both). These results suggest that a nonsteroidal anti inflammatory drug/opiate combination, together with endodontic therapy, may be useful in the management of endodontic pain. PMID- 10687524 TI - Endothelial cell adhesion molecules in human dental pulp: a comparative immunohistochemical study on chronic periodontitis. AB - Migration of leukocytes to inflammation sites through vascular endothelium is controlled by the interactions of adhesion molecules expressed on both endothelial cells and leukocytes, most of which are already covered by cluster of differentiation (CD) codes. We examined the expression of a variety of endothelial cell adhesion molecules in human dental pulp vasculature to obtain further evidence on the tissue distribution and function of these molecules by using an indirect immunoperoxidase technique. We obtained the pulp tissue samples from teeth extracted due to orthodontic reasons as controls and compared with those extracted due to chronic periodontitis. In all samples, both CD31 and CD146 were expressed by arterial, venous, and capillary endothelia. There was no significant difference between the staining intensity of normal and inflamed pulp tissues. CD102 expression on the endothelium was significantly stronger in chronic periodontitis pulp samples. CD106, CD62-E, CD62-P, CD105, and CD54 were variably expressed in control and chronic periodontitis groups. Our results indicate that CD102 represents the major endothelial cell adhesion molecule probably involved in the inflammatory reactions in chronic periodontitis. PMID- 10687525 TI - Root deformation during root-end preparation. AB - Ultrasonic root-end preparation techniques have recently been introduced and revolutionized the field of endodontic surgery. However, several reports claimed that there was an increasing incidence of crack formation after ultrasonic root end preparation. As yet, little work has focused on the root deformation during root-end preparation. Thus, the purpose of this investigation was to measure the amount of root deformation during root-end preparation with the use of microhandpiece and ultrasonic systems by using strain gauge methods, and simultaneously to detect any cracks with the aid of the stereomicroscope, stain, and an image processing system. The results demonstrated the ultrasonic instrumentation produced significantly greater strain on average than that generated with the microhandpiece system. From the viewpoint of fracture, any technique that could diminish the strain on the root would decrease the likelihood of fracture; however, no crack was observed on any resected surface of roots in this study. PMID- 10687526 TI - Evaluation of diamond-coated ultrasonic instruments for root-end preparation. AB - Ultrasonic instrumentation has been associated with cracking of the dentin in the area of the root-end preparation. The purpose of this study was to evaluate root end preparations for cracking and to describe cavosurface morphology after the use of diamond-coated instruments. Forty teeth were inspected for intradentin cracks, incomplete canal cracks, and complete canal cracks before and after preparation with a stainless steel CT-5 ultrasonic instrument and again after root-end preparation with an S12D/90 degrees diamond-coated instrument. Six teeth had polyvinylsiloxane impressions taken of the root ends after preparation with the CT-5 and again after preparation with the diamond-coated instrument. Replicas were made, split, sputter-coated, and inspected using the scanning electron microscope. This study indicates that use of the diamond-coated instrument for root-end preparation does not result in significant root-end cracking and that it can remove cracks created by a prior instrument's use. The use of the diamond coated instrument resulted in a heavily abraded, debris-covered cavosurface that may affect the apical seal. PMID- 10687527 TI - Biocompatibility of an adhesive system applied to exposed human dental pulp. AB - Human pulp tissue was directly capped with All Bond 2, or calcium hydroxide and evaluated 7, 30, or 60 days after the procedures. Histological analysis was performed to assess the inflammatory cell response, tissue disorganization, dentin bridging, and the presence of bacteria. At 7 days, with All Bond 2 capping, there was a large area of neutrophilic infiltrate underlying the pulp capping material, and the death of adjacent odontoblasts, was observed. However, with time, the neutrophilic reaction was replaced by fibroblastic proliferation with macrophages and giant cells surrounding globules of resin scattered in the coronal pulp tissue. The persistent inflammatory reaction and hyaline alteration of extracellular matrix inhibited complete pulp repair or dentin bridging. In contrast, at 7 days, the pulp tissue capped with calcium hydroxide exhibited odontoblast-like cells organized underneath coagulation necrosis. Pulp repair evolved into apparent complete dentin bridge formation at 60 days. All Bond 2 did not appear to allow any pulp repair and does not appear to be indicated for direct pulp capping of human teeth. PMID- 10687528 TI - Effect of retinoic acid on osteopontin expression in rat clonal dental pulp cells. AB - We studied the effect of retinoic acid on osteopontin synthesis and the mRNA expression in rat clonal dental pulp cells, RPC-C2A. An immunoprecipitation assay clarified that retinoic acid caused an increase in phosphorylated osteopontin synthesis that was dose-dependent, and marked increases were observed at retinoic acid concentrations of 10(-6) to 10(-5) M (1.7-fold). A Northern blotting analysis revealed a similar pattern of increase in osteopontin mRNA expression of up to 6.2-fold of control levels. Because osteopontin has an important role in the mineralization process, these results suggest that retinoic acid regulates mineralization, which takes place in the pulp cavity, including reparative dentin formation. PMID- 10687529 TI - Clinical undergraduate teaching. AB - At the present time, most endodontists who teach in the undergraduate dental school clinics do not receive formal training. Dental schools have traditionally assumed that the expert knowledge and technical skills that these endodontists could contribute would be sufficient to allow them to teach successfully. Today, we know that this is not always the case. To improve the instructional function of the dental clinics, instructors should recognize and assume their responsibilities as tone-setters, facilitators, and role models. They should define their goals and objectives clearly, and strive to create an open and trusting learning environment for their students. Instructors must also recognize the significance of their impact on the development of their students. Formal pedagogical training would assist instructors in fulfilling these responsibilities. PMID- 10687530 TI - Apical canal diameter in the first upper molar at various ages. AB - The shape of root canals cross-sectioned through their roots at 2 mm from their apices and its correlation with the D0 diameter of endodontic instruments was evaluated in 40 first upper molars. The molars were grouped according to age: under 13 yr (children), 18 to 20 yr (adolescents), 30 to 40 yr (adults), and over 50 yr. Evaluation of the root canal diameters revealed that the shapes were predominantly circular in the palatal canal, mostly flat in the mesiobuccal canal, and circular or flat in equal proportions in the distobuccal root. Age does not seem to affect the shape of the canals. Narrowing with age was statistically significant (p < 0.05) for palatal and mesiobuccal canals only. Correlation between the maximum diameter of the canals and the instruments was varied. Even in old age, diameters were observed that would require instruments of a size that would be impossible to use, because one internal diameter would exceed the root's external diameter in a different direction (i.e. intimal buccolingual diameter of #80 and external mesiodistal diameter of #70). PMID- 10687531 TI - Effect of the type carrier used on the results of dichlorodifluoromethane application to teeth. AB - Thermal testing, especially cold, is an important part of diagnosing pulpal vitality. It was the purpose of this study to determine (i) if a difference exists in temperature when Endo ice is used with four different applicators, and (ii) if there is one applicator device that provides the greatest thermal change for a tooth. Endo ice was used with the following applicators: a #2 standard size large cotton pellet, a #4 standard size small cotton pellet, a wood stick cotton tip applicator, and a cotton roll. The temperature of a mandibular incisor pulp chamber was measured in degree C 10 s after the application of the dichlorodifluoromethane (DDM) to the midfacial surface of the crown. Also, the DDM was applied to each applicator either by direct spray or by submergence in DDM liquid. This study found that the greatest temperature change was recorded by Endo Ice (DDM) sprayed directly onto a large cotton pellet. There was no significant difference between directly spraying or submerging the applicator in DDM liquid. PMID- 10687532 TI - A four-rooted quadrangular maxillary molar. AB - The endodontic treatment of a maxillary molar with an aberrant root morphology can be diagnostically and technically challenging. This case report is presented to illustrate and describe the endodontic treatment of a four-rooted maxillary molar with a quadrangular root trunk morphology. PMID- 10687533 TI - Evaluation of pigmented intraorifice barriers in endodontically treated teeth. AB - The purpose of this study was to evaluate the effectiveness of three pigmented glass ionomer cements used as intraorifice barriers to prevent coronal microleakage. One hundred ten extracted mandibular human premolars were divided into four experimental groups of 25 teeth each and two control groups of 5 teeth each. The experimental teeth were instrumented and obturated using thermoplasticized gutta-percha and AH26 sealer. Group 1 teeth received no further treatment. Teeth in groups 2 through 4 had 1 of 3 pigmented glass ionomers (Vitrebond, GC America, and Ketac-Bond) placed as an intraorifice barrier. Positive control teeth were instrumented but not obturated. The negative control teeth were instrumented, obturated, and externally sealed with epoxy resin. The coronal 3 mm of each root was sealed into the lumen of an 18-mm segment of latex surgical tubing. After the apparatus was sterilized, 2.0 ml of a 24 h growth of Proteus vulgaris in trypticase soy broth (TSB) was placed in the coronal reservoir of the tooth. The inoculated apparatus was placed into a presterilized test tube containing 1.5 ml of TSB and incubated for 90 days at 37 degrees C. The TSB in the lower reservoir was observed daily for turbidity, which would indicate leakage along the full length of the obturated root canal. To determine if differences in microbial leakage occurred among the four experimental groups, Pearson's chi 2 and Fisher's exact tests were performed. The confidence level was set at 95%. The positive and negative controls validated the microbial testing method. The teeth without an intraorifice barrier leaked significantly more than teeth with Vitrebond intraorifice barriers (p < 0.05). The difference in leakage among the experimental glass ionomer barriers was not significant (p > 0.05). PMID- 10687534 TI - Changes in root surface temperatures with in vitro use of the system B HeatSource. AB - The purpose of this study was to measure root surface temperatures while using the System B HeatSource at various temperature settings. A split-tooth model of a human maxillary central incisor was prepared with 10 thermocouples to record root surface temperatures at 1-mm increments from the root apex. A System B HeatSource model 1005 was used to warm and compact gutta-percha to within 3 mm of the working length using the Buchanan technique. Twenty obturations were recorded at each of the following temperature settings: 250 degrees, 300 degrees, 350 degrees, 400 degrees, 450 degrees, 500 degrees, 550 degrees, and 600 degrees C. Examination of the mean temperatures recorded for each position and at each temperature setting revealed that the thermocouple 5 mm from the apex (T5) detected the highest increases in root surface temperatures. Only this site exceeded the 10 degrees C rise in temperature for one full minute that could cause damage to the supporting structures. The range of instantaneous temperatures at this site was 8.85 to 12.06 degrees C, with a mean of 10.62 +/- 0.93 degrees C. The results of this in vitro study indicate that any temperature setting of the System B HeatSource at or above 250 degrees C has the potential to cause the root surface temperature to rise 10 degrees C. Whether this occurs in vivo or if it does is maintained long enough to cause any tissue damage remains to be determined. PMID- 10687535 TI - Rapid determination of dry weight in human dental pulp by a colorimetric reaction. AB - This study was designed to assess total carbohydrate (TC) concentration, as well as the noncollagenous protein content, in human dental pulp. Pulps were obtained from eight premolars (13.10 +/- 4.33 mg weight, mean +/- SD) and homogenized in saline solution. TC content was as follows: 16.68 +/- 9.49 micrograms/mg of tissue (mean +/- SD); 3.22 +/- 1.69 ng of TC/mg protein (mean +/- SD); and 16.23 +/- 6.80 micrograms of TC/mg total organic material (mean +/- SD). The high concentration of carbohydrates observed in the pulp is a result of the presence of glycoproteins, glycosaminoglycans (chondroitin-6-sulfate, heparan sulfate, hyaluronic acid, and dermatan sulfate), and proteoglycans, whose structural, biochemical, and physiological functions have been well documented. Dry weight determination using dichromate solution was used because it shows less dispersion than when the data on carbohydrates and proteins are expressed as wet weight. PMID- 10687536 TI - An improved technique for the evaluation of root canal preparation. AB - A device and a method are described for the simultaneous in vitro evaluation of several important parameters of root canal preparation, including root canal cleanliness, straightening, changes in root canal diameter, working safety (loss of working length, apical blockage, instrument fracture, and apical perforation), measurement of apically extruded debris, working time, and practicability. Using a modification of the muffle system described by Bramante et al. (J. Endodon, 13:243-5, 1987), all of these parameters can be investigated simultaneously during preparation of two mesial root canals in extracted mandibular molars. PMID- 10687537 TI - A comparative study of apical leakage of Apexit, Ketac-Endo, and Diaket root canal sealers. AB - The purpose of this study was to compare the apical leakage of Ketac-Endo, Apexit, and Diaket. Fifty freshly extracted human maxillary anterior teeth were used. The anatomical crowns were removed at the amelocemental junction, and step back preparation of the roots canals was performed with K-type files to size 40 using 2 ml of 5.25% NaOCl irrigant after each file change. The roots were randomly divided into five groups of 10 roots each: three experimental and two control groups. The experimental groups were as follows: group 1, Apexit; group 2, Ketac-Endo; and group 3, Diaket. Root canals were filled with one of the sealers and gutta-percha using lateral condensation. After the specimens were stored in 100% humidity at 37 degrees C for 2 wk, the roots were covered with two layers of nail polish and immersed in 2% methylene blue for 7 days. Each tooth was split into two sections, and dye penetration was evaluated independently by three examiners using a stereomicroscope at x20 magnification. Mann-Whitney U analysis showed that there was no significant difference between Apexit and Diaket (p > 0.05). However, there was significantly more leakage with Ketac-Endo (p < 0.05). PMID- 10687538 TI - Evaluation of microbial infiltration in restored cavities--an alternative method. AB - This work evaluated the efficacy of an improved method used to determine the frequency of bacterial infiltration and bacterial population levels and morphotypes in cavities restored with adhesive composites in conventional mice. By using the alternative methodology suggested in this work, bacteria from microleakage were recovered and identified in cavities subjected to restoration procedures that used acid etching of the dentin and dentin adhesives used with light-curing resin. The methodology presented herein seems to be more effective than the one normally used to investigate the presence of bacteria, which uses acid demineralization of dental structures for the histological processing of tissues. The results suggest that the methodology presented in this work made it possible to recover and identify Gram-negative and Gram-positive bacteria from microleakage. Frequencies of microleakage and bacterial population levels in restored cavities using two different adhesive systems were not statistically different (p < 0.05). PMID- 10687539 TI - Evaluation of apical seal in straight canals after obturation using the Lightspeed sectional method. AB - This study evaluated the sectional gutta-percha obturation technique advocated by Lightspeed Technology, Inc. Single relatively straight-canaled teeth were prepared with rotary instruments and divided into three groups. Groups A and B served as controls and were obturated using laterally condensed gutta-percha and either Roth's 801 or Ketac-Endo sealer. Group C was obturated using the Lightspeed technique that included placement of a 5-mm apical section of gutta percha, followed by backfill with Ketac-Endo sealer and a single gutta-percha cone. All teeth were suspended in India ink for 14 days then cleared. Four additional teeth, which were obturated as in group C then sectioned, revealed a tightly adapted apical section of gutta-percha with a very thin layer of sealer. However, voids were noted in the middle and coronal areas. There was no significant difference in apical microleakage among the three groups. The sectional method was significantly faster than lateral condensation and seems to offer promise as an effective obturation method. PMID- 10687540 TI - Leakage associated with single or multiple increment backfill with the Obtura II gutta-percha system. AB - The purpose of this in vitro study was to compare dye leakage between canals backfilled in a single increment and canals backfilled in multiple increments using the Obtura II system with two different sealers. Sixty extracted single canal teeth were decoronated, cleaned, and shaped. After master cone fit and placement of either Roth 801 or AH26 sealers, the canals were down-packed to 4 mm from working length. The teeth were then divided into 4 groups of 15: group 1- Roth 801 sealer with 1 increment of Obtura II back-fill; group 2--Roth 801 sealer backfilled in 4- to 5-mm increments; group 3--AH26 sealer with 1 increment of backfill; and group 4--AH26 sealer backfilled in 4- to 5-mm increments. After sealer set, the teeth were apicected 5 mm from working length. The apices were discarded. The coronal segments were coated with two layers of fingernail polish, except for the resected apical end. The teeth were immersed in Pelikan ink for 5 days. The teeth were then decalcified, dehydrated, and rendered transparent in methylsalicylate. Dye penetration was measured on four surfaces of each root. The mean measurements of each tooth were averaged for each group. Leakage of group 1 was 6.69; group 2, 5.39; group 3, 5.71; and group 4, 5.02. Differences were not statistically significant (p > 0.05). This study suggests that it may be clinically acceptable to backfill canals up to 10 mm in a single increment using sealer and the Obtura II gutta-percha system. PMID- 10687541 TI - Comparison of two implantation sites for testing intraosseous biocompatibility. AB - The purpose of this study was to compare two implantation sites--the mandible and the femur of the rabbit--for testing in vivo intraosseous biocompatility. Twenty two new Zealand rabbits were anesthetized, and the mandibular and femur bones were exposed. A hybrid glass ionomer cement or zinc oxide eugenol cement was loaded into silicone carriers and inserted into the two bones after drilling the two cortical plates. Eleven rabbits were killed 4 wk after implantation, and 11 rabbits were killed after 12 wk. The mandibles and femurs were prepared using standard histological procedures; tissue reactions were graded from none to severe. At 4 wk, no statistically significant difference was found between the two implantation sites. After 12 weeks, bone healing was statistically better in the mandible than in the femur. The mandible seems to be a better implantation site in the case of intraosseous implantation tests. The intraosseous biocompatibility of Vitremer was similar at 4 wk and superior at 12 wk to that of Super-EBA. PMID- 10687542 TI - Eosinophil-derived transforming growth factors (TGF-alpha and TGF-beta 1) in human periradicular lesions. AB - Inflammatory mediators of periradicular lesions are poorly understood. Transforming growth factors-alpha and -beta 1 (TGF-alpha and TGF-beta 1) have been linked with the cellular processes for both soft and hard tissue wound healing. The purpose of this study is to demonstrate the cellular sources of TGF alpha and TGF-beta 1 mRNA and protein in periapical lesions by in situ hybridization and immunohistochemistry. Nine periapical granulomas and nine periapical cysts were examined. TGF-alpha mRNA and protein were not detectable in the granulomas examined. However, eosinophils surrounding the periapical cysts demonstrated both TGF-alpha mRNA and protein. The vast majority of eosinophils present in the periapical granulomas and cysts also demonstrated TGF-beta 1 mRNA and protein. Other cells producing TGF-beta 1 were lymphocytes, fibroblasts, and monocytes. The presence of wound repair cytokines, such as TGF-alpha and TGF-beta 1, suggests a mechanism by which the host inflammatory response may participate in the repair and remodeling of periapical tissues. PMID- 10687543 TI - Effect of preflaring on Root ZX apex locators. AB - The Root ZX apex locator is an example of a generation of apex locators that identify the terminus of the canal by measuring a ratio between two electrical impedances. Studies have shown this device to have a high degree of accuracy. However, the manufacturer warns that the performance of these devices is limited by the presence of calcifications and dentinal shaving obstructions. An in vitro study was designed to determine if preflaring of canals would facilitate the passage of files to the apical foramen by eliminating cervical interferences and to see what effect this would have on the performance of the Root ZX apex locator. Thirty-two canals were divided into two groups. Group 1 was not manipulated before use of the Root ZX apex locator and served as control. In group 2, the canals were preflared before the use of the Root Zx apex locator. The working length files were secured in place and measured with the linear measurement tool used by the Visilog 5 imaging program. Results of this study suggest that preflaring of canals will allow working length files to more consistently reach the apical foramen (p = 0.015), which in turn increases the efficacy of the Root ZX apex locator. PMID- 10687544 TI - Effectiveness of gutta-percha removal with and without the microscope. AB - The microscope may be useful in retreatment to enhance removal of gutta-percha (GP) and to identify deficiencies in the original treatment. This study compared the effectiveness of GP removal with and without a microscope. Forty-five extracted canines were stepback prepared and obturated with GP and Roth's sealer using lateral condensation. Teeth were stored for 17 months in a humidor, then divided into four groups: group 1 (n = 20)--GP removal without aid of the microscope (group 1 criteria for GP removal was lack of GP on final files and on paper points agitated within the chloroform-filled canal); group 2 (n = 20)--GP removal with the microscope. (In addition to group 1 criteria, canals were inspected with the microscope; additional GP was removed when identified. In both groups, GP was removed using a combination of mechanical instrumentation, chloroform solvent, and K-files); group 3 (n = 3)--positive control, canals remained obturated; and group 4 (n = 2)--negative control, canals were neither prepared nor obturated. Teeth were split longitudinally, photographed, then divided into thirds to compare the apical, middle, and cervical thirds. Remnants of GP and sealer on the canal wall were traced with a digitizer, and the remaining mean % GP was analyzed by t test. Group 1 had 8.3% remaining GP, and group 2 had 7.3% remaining GP. There was no significant difference demonstrated between the two experimental groups. PMID- 10687545 TI - Dental blood supply in the segmentally resected mandible. AB - There are approximately 30,000 new cases of oral and pharyngeal carcinoma treated in the United States each year. A large number of these patients go on to receive segmental resection of the mandible, and have natural teeth remaining on the surgical side. To the best of our knowledge, there has not been a thorough discussion of the blood supply to these remaining teeth. Radiographic evidence of periapical pathology in these teeth is unusual, despite the compromised vascular supply. The purpose of this article is to report a case and review the literature on blood supply to teeth after segmental mandibulectomy. Microscopic examination was conducted on the pulpal tissue of a premolar retained on the side of, and anterior to, a segmental mandibular resection. Although abnormal, the pulp tissue showed evidence of a vascular supply 4 yr after mandibular surgery. A literature review was performed, and a discussion is given to explain the continued vascularity of the dentition through collateral and retrograde circulation. Despite the compromised dental circulation on the surgical side, unless radiographic evidence of periapical pathology occurs, endodontic therapy or extraction is not necessary. Due to the compromised nature of the circulation however, these teeth may be more susceptible to caries or restorative dental procedures that may lead to pulpal necrosis. PMID- 10687546 TI - A new approach for the retrieval of broken instruments. AB - The purpose of this article is to present a modified ultrasonic spreader and a new technique that are used for the retrieval of solid obstructions that can not be bypassed by conventional methods. The technique advocated and the instruments proposed are described. A clinical case is discussed to show the possibilities and limitations of both instrument and method. PMID- 10687547 TI - Zebra. XVII. Part 2: The case of the elusive gutta-percha. PMID- 10687548 TI - Adaptive skin blood flow increases during hip-down lying in elderly women. AB - OBJECTIVE: Pressure ulcer development due to unrelieved pressure during extended cardiovascular, orthopedic, and other procedures is an important clinical problem. Because blood flow changes within pressure-loaded tissue affect the skin breakdown process, the relative effects of 2 support surface strategies on trochanter skin blood flow were investigated. DESIGN: Skin blood perfusion was assessed by laser Doppler methods during 1 hour of continuous loading. Blood perfusion was measured before and during hip-down loading on a gel pad (static surface) and a dynamic multisegmental surface that provided periodic alternating pressure relief. Female volunteers (N = 20, age > or = 60 years) were tested on each surface in random order with sequential tests separated by 5 to 8 days. Effects were assessed by comparing perfusion during the first and last 15 minutes of hip-down loading with a 15-minute baseline. SETTING: Research center. RESULTS: Pre-load perfusions (dynamic vs static support) were similar (0.57 +/- 0.06 vs 0.64 +/- 0.08). During loading, however, a significant progressive increase in perfusion was noted only with dynamic support; by the end of the loading interval, this increase in perfusion had significantly exceeded the pre-load baseline (1.22 +/- 0.26, P = 0.001). CONCLUSION: These findings reveal a surface dependent blood flow impact, with the multisegmental dynamic approach being associated with greater flow during loading. The mechanism, though speculative, is consistent with a greater vascular adaptation potential offered by the dynamic surface. Conditions that facilitate such adaptive flow increases would appear to be of considerable benefit in helping to prevent ulcer development. PMID- 10687549 TI - Why wound care testing? PMID- 10687550 TI - The national examination blueprint: information and practice questions. PMID- 10687552 TI - Prophylactic foot surgery in the diabetic patient. AB - The preoperative and perioperative evaluation of diabetic foot pathologies has been discussed. The effects of vascular and neuropathic alterations in such cases can be devastating. Only through understanding of the underlying mechanisms that contribute to the diabetic foot can progressive, appropriate care be rendered. PMID- 10687551 TI - History of the American Academy of Wound Management. PMID- 10687553 TI - The role of biotechnology in the global economy. PMID- 10687555 TI - Consensus development conference on diabetic foot wound care. 7-8 April 1999, Boston, MA. American Diabetes Association PMID- 10687554 TI - Pressure ulcers in community-based older adults receiving home health care. Prevalence, incidence, and associated risk factors. AB - OBJECTIVES: To determine the prevalence and incidence of pressure ulcers in community-based adults receiving home health care and to identify risk factors for incident Stage II to IV pressure ulcers. DESIGN: Retrospective cohort study. SETTING: A large midwestern urban home health care agency. PATIENTS: The study cohort was 1711 nonhospice, nonintravenous therapy subjects admitted between January 1995 and March 1996 who were > or = age 60 and pressure ulcer-free on admission. MEASUREMENTS: Data on risk factors were extracted from admission information. Patient records were followed forward chronologically to the outcomes: pressure ulcer development or no pressure ulcer. MAIN RESULTS: The incidence of Stage II to IV pressure ulcers was 3.2%. Cox regression analyses revealed that limitation in activity to a wheelchair, needing assistance with the activities of daily living--dressing, bowel and/or bladder incontinence, a Braden Scale mobility subscore of very limited, anemia, adult child as primary caregiver, male gender, a recent fracture, oxygen use, and skin drainage predicted pressure ulcer development (P < or = 0.05) in this exploratory model. CONCLUSIONS: Patients > or = age 60 who are admitted to a home health care agency with 1 or more of these risk factors require close monitoring for pressure ulcer development and should be taught preventive interventions on admission. PMID- 10687556 TI - Complementary and alternative medicine in wound healing. PMID- 10687557 TI - Off-loading techniques in the treatment of diabetic plantar neuropathic foot ulceration. AB - In the individual with diabetes mellitus, foot ulceration represents the single most important risk factor in lower-extremity amputation. The goal of treatment is to obtain a healed and closed wound that (1) eliminates a portal of entry for bacterial invasion and development of limb-threatening infection, and (2) allows for tissue loading. This manuscript reviews current off-loading approaches to the treatment of plantar neuropathic foot ulcers, along with advantages and disadvantages of those techniques. PMID- 10687558 TI - Report on the prevalence of skin ulcers in a home health agency population. AB - OBJECTIVE: This survey was conducted to assess the presence of skin ulcers within a home health agency population in the United States. DESIGN: This voluntary survey was conducted by 177 home health agencies. A single observation of each patient within the agency's active caseload formed the cohort examined. Patients deemed to be at low risk (Braden Scale score > 19) were eliminated from further evaluation, while those with skin ulcers were evaluated for wound- and caregiver related factors. Surveys were conducted between March 1, 1996, and December 31, 1997. SETTING: Home health agencies in 19 states throughout the United States, with no restrictions on the type or acuity of the patients served. RESULTS: A total of 21,529 patients were surveyed, with a prevalence of pressure ulcers (inclusive of all stages) of 6.8% (n = 1455). Rates for each agency ranged between 0.5% and 35.7%. The total number of ulcers reported was 2526 (average per patient was 1.7), with 36% (n = 919) found on the sacrum and the buttocks. CONCLUSION: Pressure ulcers were the most frequently reported reason for admission to the agency's caseload. Survey results are similar to rates reported in other segments of the health care industry. However, among the home health care population, the primary caregiver is unlikely to be a health care professional. This survey found that the patient's spouse was the primary caregiver in 30% (n = 437) of the 1450 responses received regarding the relationship of the primary caregiver to the patient. PMID- 10687559 TI - Assessment and diagnosis of burn wounds. PMID- 10687560 TI - The APTA electrical stimulation lawsuit and its aftermath. American Physical Therapy Association. PMID- 10687561 TI - Misunderstood illnesses: fibromyalgia and chronic fatigue syndrome. PMID- 10687562 TI - Perspective: why I volunteer. PMID- 10687563 TI - Nurses as a scarce resource. PMID- 10687564 TI - Health Professions Act--what are the gains for the nursing profession? PMID- 10687565 TI - Job action and the code of ethics. PMID- 10687566 TI - Remembering contributions of nurses during WWII. PMID- 10687567 TI - Standards for registered nurses. Alternate and complementary therapy in nursing practise. PMID- 10687568 TI - Whither cesareans in the new millenium? PMID- 10687569 TI - Knee-chest postural management for breech at term: a randomized controlled trial. AB - BACKGROUND: In 3 to 4 percent of all term births, the fetus presents as a breech. The objectives of this trial were to assess if assuming the knee-chest position reduced the frequency of breech presentation at delivery, increased the success of the subsequent external cephalic version, or both, and to determine if this management plan reduced the need for cesarean delivery. METHODS: A randomized clinical trial recruited 100 women from two hospitals in Adelaide, South Australia, with a singleton breech presentation and a gestational age equal to or more than 36 weeks. Women in the treatment group were advised to assume the knee chest position for 15 minutes three times a day for one week. Women in the control group did not perform postural management. All participants were reviewed one week later, and women whose baby remained as a breech presentation were offered an external cephalic version. RESULTS: Postural management did not increase the success of the external cephalic version, reduce the frequency of breech presentation at delivery, or reduce the need for cesarean delivery in women with a breech presentation at term. CONCLUSIONS: Findings from this trial included in a meta-analysis of postural management for breech presentation at term suggested that this is not an effective form of care to be offered routinely to women with a breech presentation at term. PMID- 10687570 TI - Smoking relapse and early weaning among postpartum women: is there an association? AB - BACKGROUND: Smoking in the postpartum period may contribute to early weaning, although the nature and temporal aspect of the relationship are poorly understood. The objective of this study was to examine the association between early weaning and smoking relapse among women who stopped smoking during pregnancy. METHODS: A secondary analysis of data from a randomized controlled trial was conducted. The participants were 228 women who had stopped smoking for pregnancy, who participated in a smoking relapse prevention trial, and who breastfed. Women who relapsed to daily smoking postpartum were compared with those who remained abstinent or smoked occasionally. The dependent variable was breastfeeding for less than 26 weeks (early weaning). Potential covariates included intended duration of breastfeeding, parity, partner's smoking, nicotine dependence, emotional health, return to paid employment, and various sociodemographic variables. RESULTS: Approximately two-thirds (65.1%) of the women who relapsed to daily smoking weaned before 26 weeks compared with 33.8 percent of the women who remained abstinent or smoked occasionally. Controlling for intended duration of breastfeeding, education, and return to paid employment, women who resumed daily smoking were almost four times more likely to wean early than those who abstained or smoked occasionally. CONCLUSIONS: Early weaning may result from psychological or physiological changes associated with tobacco use. Smoking relapse prevention in the postpartum period may be one of the most effective interventions in ensuring that women who stop smoking for pregnancy remain stopped and breastfeed their babies for the recommended duration. PMID- 10687571 TI - Effect of labor analgesia on breastfeeding success. AB - BACKGROUND: The effect of labor analgesia on breastfeeding success is not well defined. Some authors have hypothesized that labor analgesia may affect lactation success. The purpose of this observational study was to determine if intrapartum analgesia influenced breastfeeding success at 6 weeks postpartum in a setting that strongly supported breastfeeding. METHODS: Healthy women with uncomplicated term pregnancies who planned to breastfeed consented to a telephone interview. We recorded demographic data, labor induction status, delivery mode, and analgesic medications. At between 6 and 8 weeks postpartum, patients were asked to describe breastfeeding use, problems encountered, solutions derived, sources of support and information, and satisfaction. We created a logistic regression model using intrapartum analgesia information and controlling for demographic factors previously correlated with lactation success. RESULTS: We enrolled 189 women, contacted 177 women postpartum, and obtained complete data on 171 women. Of these, 59 percent received epidural analgesia, 72 percent breastfed fully, and 20 percent breastfed partially (> 50% of infant nutrition) at 6 weeks postpartum. After controlling for demographics and labor outcome, we could not demonstrate a correlation between breastfeeding success at 6 to 8 weeks and labor analgesia. CONCLUSIONS: In a hospital that strongly promotes breastfeeding, epidural labor analgesia with local anesthetics and opioids does not impede breastfeeding success. We recommend that hospitals that find decreased lactation success in parturients receiving epidural analgesia reexamine their postdelivery care policies. PMID- 10687572 TI - Managing labor using partograms with different action lines: a prospective study of women's views. AB - BACKGROUND: The precise timing of medical intervention for women in prolonged labor is the subject of considerable debate. The partogram action line is a tool to assist practitioners in the correct diagnosis of prolonged labor. Despite its widespread use, the precise timing of the action line has not been rigorously studied, and women's views have rarely been sought. The aim of this study was to assess the effect on maternal satisfaction of managing labor using partograms with action lines drawn at 2, 3, or 4 hours to the right of the alert line. METHODS: As part of a large pilot randomized controlled trial, women's views were explored using a specifically designed questionnaire that was completed by 615 primiparas 2 days after giving birth. The quantifiable data were analyzed by comparing means using ANOVA followed by the Scheffe test. RESULTS: Women in the 2 hour arm were significantly more satisfied than those in the other two arms (p < 0.001), despite having the most obstetric intervention. CONCLUSIONS: For women in prolonged labor, obstetric intervention can be an acceptable or even favorable option. Midwives and obstetricians need to provide labor management that takes into account the preferences of the women to whom they give care. PMID- 10687573 TI - Commentary: managing labor: what do women really want? PMID- 10687574 TI - Women's perceptions of midwifery care: a longitudinal study to shape curriculum development. AB - BACKGROUND: Health and education services are increasingly expected to focus on the consumer. The perceptions of childbearing women should be incorporated into midwifery curricula, but often they are given minimal attention or not sought for this purpose. This study was designed to enable the views and experiences of local women to influence curriculum development in a large university in England. METHODS: A descriptive, longitudinal, qualitative study was conducted using semistructured and unstructured interviews with women, and data from their maternity records. Forty-one pregnant women were recruited and interviewed during pregnancy, in the early postpartum period in hospital, and in their homes 2 to 3 weeks after the birth. RESULTS: Themes were clustered into three categories: the characteristics and qualities of the caregivers, the individualized nature of care, and the clinical competence of the caregivers. Continuity of caregiver was desired but accepted as probably unrealistic by many. Developing a "special" trusting relationship with a female midwife was perceived as essential to promoting a positive childbirth experience. Clinical competence was expected and largely experienced. Negative feelings related to individual caregivers more than the type of care given. CONCLUSIONS: Most women had positive experiences, finding midwives and doctors with good knowledge, interpersonal skills, and abilities. Examples of poor communication skills and interprofessional conflict indicated a need to give priority to developing and assessing students' interpersonal skills. Evidence of interprofessional conflict acted as the catalyst to merge midwifery with the department of obstetrics in the university to enhance interprofessional learning. PMID- 10687575 TI - Severity of nausea and vomiting during pregnancy: what does it predict? AB - BACKGROUND: Relationships between the severity of nausea and vomiting during pregnancy and selected demographic (employment status, parity, age, smoking) and pregnancy outcome (birthweight, gender) variables are described. METHODS: Women who volunteered for a community-based clinical trial were eligible for inclusion in this study. On three occasions, 12 hours apart, during early pregnancy using a continuous measure of nausea, vomiting, and retching, women assessed the amount, duration, and severity of symptoms as they occurred. After the birth of their infants, they provided information about the duration of nausea, vomiting, and retching as well pregnancy outcome information by responding to a mailed questionnaire. Multivariate methods were used to analyze data. RESULTS: More severe vomiting in early pregnancy was likely to continue for a longer period of time and was related to decreased infant birthweight. Gestational age, parity status, and severity of vomiting were predictors of infant birthweight and together explained 22 percent of the variance in birthweight. A significant relationship between fetal gender and severity of nausea and vomiting was not found. CONCLUSIONS: It may be possible to identify women at risk for third trimester vomiting and to provide appropriate nutritional support and counseling so that their risk of having a low-birthweight infant is reduced. A larger sample would be required to assess the relationship between fetal gender and severity of nausea, vomiting, and retching. PMID- 10687576 TI - A comparison of breast stimulation and intravenous oxytocin for the augmentation of labor. AB - BACKGROUND: Breast stimulation to augment labor has been used for centuries in tribal societies and by midwives. In recent years it has been shown to be effective in ripening the cervix, inducing labor, and as an alternative to oxytocin for the contraction stress test. This study compared the effectiveness of breast stimulation with oxytocin infusion in augmenting labor. METHODS: Women admitted to the labor ward were eligible for the study if they had inadequate labor with premature rupture of the membranes and met inclusion criteria. They were assigned to oxytocin augmentation or breast stimulation (manual or pump), and were switched to oxytocin in the event of method failure. Outcomes included time to delivery, intervention to delivery, proportion of spontaneous deliveries, and Apgar scores. One hundred participants were needed in each arm of the study to demonstrate a 2- to 3-hour difference in delivery time, with a power of 80 percent. RESULTS: Analysis was performed on 79 women, of whom 49 were in the breast stimulation group and 30 in the oxytocin group. Sixty-five percent of the participants failed breast stimulation and were switched to oxytocin infusion. Although augmentation start to delivery was shorter for the oxytocin group (p < 0.001), no differences in total labor time occurred between the groups. Nulliparas receiving breast stimulation had more spontaneous (relative risk 1.7, p = 0.04), and fewer instrumental deliveries than those receiving oxytocin (relative risk 0.2, p = 0.02). No significant differences in adverse fetal outcomes occurred between the study groups. CONCLUSIONS: The small number of participants and a variety of problems with the conduct of the study prevented the formulation of reliable conclusions from the results. However, the study provided important insights into the feasibility and problems of developing a high-quality randomized trial of augmentation by breast stimulation. PMID- 10687577 TI - Breast stimulation to augment labor: history, mystery, and culture. AB - This paper describes the development of a researchable project, arising from the clinical observation of a physiologic phenomenon during labor. Augmentation of labor by breast stimulation has been used in a variety of cultures for centuries. The process of developing a clinical study of augmentation in the modern obstetric environment is discussed, with reference to cultural attitudes of patients and health care workers. PMID- 10687578 TI - Does walking enhance labor progress? PMID- 10687579 TI - Measurement of distress. A necessity for all patients. PMID- 10687580 TI - Woman with lung cancer who continues to smoke. PMID- 10687581 TI - Age, self-efficacy, and change in patients' adjustment to cancer. AB - OBJECTIVES: The purpose of this secondary analysis was to investigate cancer patients' self-care self-efficacy and measures of adjustment over time, as well as the role of self-care self-efficacy with measures of adjustment. The primary study was a longitudinal study of cancer patients and family members' adjustments. MATERIALS AND METHODS: Three hundred seven cancer patients at all stages of cancer completed study instruments on one occasion; 181 completed the instruments 4 months later; and 124, 8 months later. Instruments included the Strategies Used by Patients to Promote Health (SUPPH) to measure self-care self efficacy, the Functional Assessment of Cancer Treatment (FACT) to measure quality of life, the Profile of Mood States (POMS) to measure patients' mood disturbances, and the Symptom Distress Scale (SDS) to measure patients' concerns. RESULTS: Using analysis of variance, a series of one-way repeated measures used to investigate changes in cancer patients' self-care self-efficacy and measures of adjustment revealed significant decreases in patients' self-care self-efficacy (P = .01) and quality of life (P = .001) over time. Patients' symptoms and mood disturbances did not significantly change over time. The role of self-care self efficacy with measures of adjustment was investigated using canonical correlations. For the predictor variables, subscores for coping and enjoying life on the SUPPH showed the strongest loadings, 0.83 and 0.94, respectively. On the dependent variables, the FACT was by far the most important variable, with a loading of 0.92. CONCLUSIONS: Results demonstrate that without intervention, cancer patients' measures of self-efficacy and adjustment decrease over time, and patients' self-efficacy influences their adjustment. Psychosocial interventions have been designed to increase self-efficacy and to enhance adjustment. Longitudinal study of efficacy-enhancing interventions is needed. PMID- 10687582 TI - The effect of Epoetin alfa on quality of life in anemic cancer patients. AB - OBJECTIVES: The purpose of this paper is to review evidence on the use of Epoetin alfa in the treatment of anemia associated with cancer treatment through a discussion of clinical and quality-of-life considerations. MATERIALS AND METHODS: Cancer patients often experience fatigue, which leads to reduced quality of life. There are few effective treatments available to manage this potentially debilitating symptom, which can lead clinicians to become discouraged about treating fatigue. When cancer-related fatigue is due to anemia, there are viable treatment options. This paper reviews the current management of anemia in cancer patients, with an emphasis on the use of recombinant human erythropoietin (Epoetin alfa). CONCLUSIONS: Anemia can contribute to the reduced quality of life experienced by cancer patients. Blood transfusion, the traditional method of treating anemia, is effective and relatively inexpensive, but is associated with certain risks and is subject to limitations in blood supply. Epoetin alfa therapy provides healthcare providers with an effective alternative to blood transfusion, and trial results suggest that this intervention has a positive effect on patients' quality of life. The optimal starting and stopping points for therapy have not yet been determined. In practice, many physicians begin treatment when hemoglobin levels drop below 10 g/dL and stop when they rise above 13 g/dL, with a 75% dose reduction until completion of chemotherapy if hemoglobin again drops below 12 g/dL. Nonresponse (< 1 g/dL rise in hemoglobin) is met with dose increase at 4 weeks and discontinuation after 8 weeks. Controlled studies comparing anemia management via transfusion to the use of Epoetin alfa have not been done to date. Therefore, the relative cost-effectiveness of Epoetin alfa, an effective but expensive intervention, remains unknown. PMID- 10687583 TI - Healing Icons: art support program for patients with cancer. AB - OBJECTIVES: The purpose of this report is to describe the structure and process of an art support program for patients with cancer who are age 16 and older. MATERIALS AND METHODS: Healing Icons is a six-session art support program for cancer patients. During the program participants create a three-dimensional mixed media art piece to convey a unique personal perspective on receiving a diagnosis of and being treated for cancer. Concurrently, the patients spontaneously share common experiences about their cancer, which leads to strong emotional bonds. The purpose and goals of the program, method of implementation, and evaluation are described. Information and suggestions that clinicians might find useful in developing similar programs are discussed. Patient participants, their families, and staff in the cancer center have reported positive clinical evaluations. CONCLUSIONS: The benefits of Healing Icons are derived from the therapeutic factors present in a traditional support group blended with the creative process. This kind of program opens new avenues for expressing feelings and thoughts but should be structured in such a way that group processes are not allowed to negatively impact participants. Healthcare professionals interested in collaborating with artists on similar programs for cancer patients may approach artists through local art councils, art schools, and artists guilds. Brainstorming sessions with artists would help to capitalize on the expertise of artists within the community. Initiating a pilot project would help gauge patient interest and would provide valuable feedback from the healthcare team. Research is needed to validate the clinical outcomes derived from this program, as empirical findings would greatly enhance the clinical evaluations. PMID- 10687584 TI - Reintegration after bone marrow transplantation. AB - OBJECTIVES: This study examines the problems of bone marrow transplantation (BMT) survivors in returning to "normal" life in the community after BMT. MATERIALS AND METHODS: Before being released from The Johns Hopkins Oncology Center, 84 recipients of BMT were interviewed regarding their quality of life and psychosocial adaptation. Survivors were reinterviewed at 6 months, and at 1 year post-BMT, producing considerable qualitative data regarding their problems in living. Eighty-four patients who had received BMT completed qualitative interviews and standardized measures before treatment, before the return home, and at 6 and 12 months post-BMT. The interviews were subjected to a content analysis methodology to establish units and categories to examine the body of material. RESULTS: Content analysis of these interviews from the first year after BMT identified three areas of psychosocial morbidity; 1) physical problems, which included fatigue, appearance, troubles in eating, and physical restrictions; 2) psychological problems, which included fears about the future, sense of loss of control, anxiety, and depression; and 3) community reintegration problems, which included difficulty in returning to former social roles, separation from home, family, and friends, difficulty in resuming social relations, dealing with stigmatization, problems with family and children, and financial and employment difficulties. CONCLUSIONS: Identification of these problems for BMT survivors can be used to guide the development of specific materials and services to prepare recipients of BMTs and their families for life after the transplant. These qualitative results can also be used to direct the development of assessment tools to identify potential patient and family problems. PMID- 10687586 TI - The low-bacteria diet for immunocompromised patients. Reasonable prudence or clinical superstition? PMID- 10687585 TI - Findings from an educational support course for patients with leukemia. AB - OBJECTIVES: Recent evidence indicates that patients with leukemia are a distinct subset of cancer patients with specific adjustment issues and special needs for support and follow-up. This article shares recent research findings on an Australian educational support course, appropriately named Taking Control, designed specifically for patients with leukemia and associated disorders. MATERIALS AND METHODS: The material presented in this article represents the findings from the retrospective arm of a descriptive study designed to evaluate this course. Participants in the course during the calendar year of 1997 were surveyed with an author-designed, self-report questionnaire requesting feedback on their experience of the course. RESULTS: Although the majority of participants were seeking information, there was strong evidence that the provision of information needs to be coupled with an understanding of the psychosocial reasons that motivated individuals to attend the course. The findings indicate that the course is perceived by participants to be an effective psychosocial intervention for assisting patients and their families cope with the serious diagnosis of leukemia. CONCLUSIONS: Even though information seeking was an important reason for attending this educational course, there were many other significant emotional concerns that individuals brought to the experience. When exposed to the stress of leukemia and its treatment, patients and their significant others may become preoccupied with information gathering. Therefore, it is important to not only explore the issue of the informational needs of participants, but also to examine the emotional needs they associate with this information gathering. The hope and expectation about sharing the findings of this program is that it will assist with the development of similar programs elsewhere and will stimulate further research on the importance of educational support groups in oncology. PMID- 10687587 TI - Support groups for children of patients with cancer. PMID- 10687588 TI - Sparfosate. A novel biomodulator of 5-fluorouracil. PMID- 10687589 TI - Protection of human subjects. Time for reassessment. PMID- 10687590 TI - Language barrier and psychiatric disorder as challenges to effective pain management. PMID- 10687591 TI - Factors hindering patients' use of medications for cancer pain. AB - OBJECTIVES: The purpose of this study was to explore the reasons that cancer patients with pain find it difficult to adhere to analgesic therapy. MATERIALS AND METHODS: Twenty-one patients with advanced cancer with pain were interviewed using a semistructured schedule of questions. Participants were asked to describe their decision making regarding analgesics and the factors that made it difficult for them to take analgesics prescribed for their pain. They also were asked to describe their relationships with their healthcare providers. Themes were identified and refined using qualitative analytic techniques. Two investigators independently coded all data to ensure that findings accurately reflected participants' experiences. RESULTS: Findings reveal several factors that hindered analgesic use and the specific ways in which patients evaluated these factors in making decisions about taking pain medication. The provider-patient factors that impeded analgesic use also were described. Finally, the common use of nonpharmacologic methods of pain control offers insight into the role of these therapeutic strategies in achieving pain relief and decreasing analgesic use. CONCLUSIONS: The findings underscore the importance of early intervention to address barriers to analgesic use. Some barriers may be overcome through educational efforts. The findings suggest, however, that consistent, repeated patient education often may not be sufficient to subdue patients' negative thoughts about taking the medication. Other approaches, such as changing medications or assisting the patient to use nonpharmacologic pain strategies, may prove more successful. PMID- 10687592 TI - Screening to predict complicated grief in spouses of cancer patients. AB - OBJECTIVES: Grief is the expected reaction to the death of a family member or close friend and is accompanied by substantial distress for almost everyone who experiences it. For some the grief response becomes complicated. This pilot study sought to identify individuals at high risk for complicated grief, by 1) examining the relationships that exist between family functioning before the death, psychological distress, and the grief reaction of a family after the death, and 2) presenting the use of screening with standardized measures to identify those at risk. MATERIALS AND METHODS: This pilot study examined the relationships between family functioning, psychological distress, and grief reaction. A cross-sectional design was used and the instrument included the Family Adaptability and Cohesion Evaluation Scale (FACES III), the Brief Symptom Inventory (BSI), and the Texas Revised Inventory of Grief (TRIG). Significant relationships were identified between the level of family functioning, psychological distress and grief reaction. Depression, anxiety, and general distress were significantly correlated with the two subscales of the TRIG. CONCLUSIONS: The findings clearly illustrate the merit of psychosocial screening of spouses and suggest the possible benefits of screening before the patient's death, using FACES III and the BSI to identify which spouses are at risk for complicated grief reactions. PMID- 10687593 TI - Fatigue, pain, and depression in pre-autotransplant breast cancer patients. AB - OBJECTIVES: The purpose of this study was to determine the influence of fatigue, pain, and depression on health status in breast cancer patients who had completed adjuvant chemotherapy and were scheduled for autologous bone marrow/peripheral blood stem cell transplant (AT). MATERIALS AND METHODS: A predictive, correlational design was used. A convenience sample of 127 women with stages II, III, and IV breast cancer was recruited. The setting was an urban National Cancer Institute-designated comprehensive cancer center located in the Eastern United States. Standardized questionnaires and the Gaston-Johansson Painometer (POM) were used to measure the variables. The subjects completed questionnaires in the outpatient clinic. Relationships between the multiple dimensions of fatigue and pain, depression, and health status were examined. Hierarchical regression techniques were used to determine the variance in health status accounted for by fatigue, pain, and depression. RESULTS: The subjects were age 22 to 60 years (Mean = 45; SD = 7.6), and primarily were married, white, Protestant, college educated, employed in a professional position, and had an average yearly household income of equal to or greater than $50,000. All subjects had previously received surgery and chemotherapy. Ninety-one percent of the participants reported fatigue as measured by the Fatigue Visual Analogue Scale. Forty-seven percent of the participants reported pain as measured by the Gaston-Johansson POM visual analogue scale. Fifty-four percent of the participants reported depression, ranging from mild to severe/high. Subjects reported a mean total perceived health status rating of 50.73 (SD 10.79). Fatigue, pain, and depression were all significantly correlated to each other and to total health status. Depression (P < .001) and pain (P < .01) significantly accounted for 64% (adjusted R2 = .60) of the variance in total health status. Fatigue (P < .05) and depression (P < .001) accounted for 42% (adjusted R2 = .36) of the variance in the perception of health status. CONCLUSIONS: Women with breast cancer previously treated with chemotherapy and awaiting AT may experience fatigue, pain, depression, and alterations in health status. Pain and depression had a significant impact on a woman's total health status, whereas depression and fatigue had an influence on perceived health status. Of the different dimensions of health status, one's perceptions of health status had the strongest correlation to total health status (r = .84, P < .001). Healthcare professionals need to be aware of the effects of multiple symptoms on health status and to provide appropriate care to alleviate them. PMID- 10687594 TI - Parents' perceptions of randomization in pediatric clinical trials. Children Cancer Group. AB - OBJECTIVES: The purpose of this study was to investigate parents' knowledge and perceptions about randomization in clinical trials for children with cancer, and to determine whether parents' decisions were influenced by demographic factors, randomization circumstances, the clinical characteristics of the child with cancer, or a combination. MATERIALS AND METHODS: This study collected information from 192 parents of patients with various forms of childhood cancer who either accepted or refused randomization. A comparative case-control design was used. The Clinical Investigation Randomization Scale was administered to all participants. This scale included 32 questionnaire items (QIs) pertaining to randomization as well as a mixture of open-ended questions to obtain information about demographic and other factors. RESULTS: A predictor model was developed that accurately predicted acceptance or refusal of randomization 87% of the time. Demographic information was found to have less influence than expected on parents' decisions regarding randomization. Knowledge deficits were found among both groups of parents, those who accepted and those who refused randomization. CONCLUSIONS: What most distinguished parents who refused from those who accepted randomization was not their knowledge and information about randomized clinical trials. By far, the majority of QIs that accurately predicted acceptors and refusers involved parents' beliefs, values, and perceptions. Further research is needed to determine interventions that may enable the healthcare team to provide information and decisional support most effectively to improve the informed consent process. PMID- 10687595 TI - Patient and caregiver perceptions of cancer pain control. AB - OBJECTIVES: This study measured the perceptions of cancer patients and caregivers in Utah concerning knowledge about and adequacy of pharmacologic cancer pain control. MATERIALS AND METHODS: A descriptive survey was sent to a stratified random sample of adult cancer patients obtained from the Utah Tumor Registry. Questionnaires asked cancer patients and caregivers about their knowledge of pain control and about perceptions of the adequacy of pharmacologic cancer pain management. RESULTS: The study had a 52% response rate (259 of 500) after two mailings. Eighty-five percent (219 of 259) of the respondents stated that they had no cancer pain. With the first mailing, a "no pain" response was not offered as an option. When the researchers realized that this might be a possible response, a second mailing was sent, which may be the reason for the high response rate. CONCLUSIONS: Cancer literature indicates that much cancer pain is not effectively controlled. The majority of the respondents of this study reported no pain. Because this result is different than that reported in the literature, it may indicate that education of healthcare providers, patients, and families can improve cancer pain management and control. It may also indicate an inability of the study to obtain data from those patients having cancer pain. This study should be repeated with a focused population of advanced stage cancer patients with types of cancer typically producing high levels of cancer pain. PMID- 10687596 TI - Educational needs related to complementary and alternative therapies. PMID- 10687597 TI - The Internet. Changing the way cancer survivors obtain information. PMID- 10687598 TI - Taxanes in hormone-refractory prostate cancer. PMID- 10687599 TI - Nursing shortages threaten patient care. PMID- 10687600 TI - Getting a job as a research nurse. PMID- 10687601 TI - The reality of nursing education in the '90s. PMID- 10687602 TI - Students helping students. PMID- 10687603 TI - Workforce planning essential. PMID- 10687604 TI - Linking nurse prescribing and advanced practice. PMID- 10687605 TI - Future nursing realities. PMID- 10687606 TI - Working with a team. PMID- 10687607 TI - Commentary: orchestrating a complex situation. PMID- 10687608 TI - The rewards of being a caregiver. PMID- 10687609 TI - Homecare workers undervalued. PMID- 10687610 TI - Undergoing hyperbaric oxygen therapy. PMID- 10687611 TI - Raising awareness of legal risks. PMID- 10687612 TI - Working for positive change. PMID- 10687613 TI - New therapies enhance patient care. PMID- 10687614 TI - Onus on nurses to practise safely. PMID- 10687615 TI - Recapturing the essence of nursing. Interview by Teresa O'Connor. PMID- 10687616 TI - Fulfilling a dream. Interview by Anne Manchester. PMID- 10687617 TI - Healing touch benefits patients. Interview by Anne Manchester. PMID- 10687618 TI - The impact of the new right on health care. PMID- 10687619 TI - Pondering nurses' industrial future. PMID- 10687620 TI - Devastating illness paralyses body. AB - A patient diagnosed with Guillain-Barre syndrome discovered most health professionals knew little about the disease. He wrote this article in September last year--11 months after the onset of his illness. PMID- 10687622 TI - Report highlights medical dilemmas. PMID- 10687621 TI - The joy and privilege of caring. AB - Some patients have a profound effect on nurses. One nurse recalls the joy and privilege of caring for a much-loved old woman. PMID- 10687623 TI - Analyzing the Mental Health Act. AB - The Mental Health Act has brought significant changes to clinical practice. Here one nurse examines some of these changes and some of the difficulties nurses have in working under the Act. PMID- 10687624 TI - Paying tribute to Maori values. PMID- 10687625 TI - Understanding what makes people tick. Interview by Anne Manchester. PMID- 10687626 TI - Youthful challenges. Interview by Teresa O'Connor. PMID- 10687627 TI - Managing professional boundaries. PMID- 10687628 TI - Advancing nursing education. PMID- 10687629 TI - Making partnership a reality. PMID- 10687630 TI - A window on mental health nursing. PMID- 10687631 TI - On the road to openness. PMID- 10687632 TI - Nurses in waiting. PMID- 10687633 TI - Losing patience. PMID- 10687634 TI - Nursing by numbers. PMID- 10687635 TI - The big issue. Interview by Jenny Knight. PMID- 10687636 TI - Cuddle mums. PMID- 10687637 TI - Acting on health. PMID- 10687638 TI - Meet nurse perfect. PMID- 10687639 TI - Responding to disability. PMID- 10687640 TI - Nurses' attitudes to the extension and expansion of their clinical roles. AB - Nurses are increasingly being asked to extend or expand their traditional roles, often for reasons other than their own professional development. This study, across three specialties in one hospital, examines whether or not nurses view such change in a positive light. PMID- 10687641 TI - Improving paediatric diabetes care. AB - The authors review the management of paediatric patients in diabetic ketoacidosis. Paying particular attention to the pathophysiology of the illness and nursing documentation, they have developed a new diabetic ketoacidosis flow chart to improve nursing care. PMID- 10687642 TI - Quality and the new NHS. AB - In this article, the authors explore the concept of quality in health care and examine in detail some of the methods currently being used to assess and improve it. PMID- 10687643 TI - Holistic care for a man with prostate disease. PMID- 10687644 TI - The bank for you. PMID- 10687645 TI - Rights and responsibilities. PMID- 10687646 TI - The new Japanese Society of Pressure Ulcers: an opportunity for international collaboration. PMID- 10687647 TI - Update: SNF PPS and consolidated billing. AB - Wound, ostomy, and continence specialists have a unique opportunity for developing programs in the long-term care environment. Their expertise and perspective can assist in providing care within the PPS payment rate, improving the fiscal stature of the facility, improving patient outcomes, and increasing the facility's referral base. The HCFA has posted a tool on its website that allows the user to follow RUG III grouper logic manually. It walks through each step of the process and is designed to be a helpful learning tool: http:@www.hcfa.gov/medicare/hsqb/mds20/>. PMID- 10687649 TI - Facilitating international wound care communication and sharing. PMID- 10687648 TI - Issues of euthanasia. AB - The debate surrounding euthanasia may have existed before recorded history, yet it continues today. Advanced technology, with its implication on prolonging life and postponing death in some cases, further complicates the debate. This column includes a historical review of euthanasia, related terminology, and proposed guidelines. The implications of recent legislation on enhancing discussions around terminal care are also reviewed. PMID- 10687650 TI - Downloading plantar foot pressures in the diabetic patient. AB - Pressure downloading (offloading) is the most important component in the prevention and treatment of diabetic foot ulcers because peripheral neuropathy is a major contributing factor to more than 90% of all diabetic foot ulcers. Downloading techniques range from the simplest insole, through many types of orthotics and footwear modifications including the ankle-foot orthosis and total contact casting, to surgical procedures. A philosophical difference exists between surgical and nonsurgical approaches, with the patient subjected to the bias of the practitioner. This article explores uniting both surgical and nonsurgical pressure downloading techniques, using a modified Carville Classification System to help the practitioner determine the appropriate method(s) of downloading. By adding a Category 4 to include diabetic persons with foot ulcers or acute Charcot events, a seamless system is obtained to categorize and treat all people with diabetes with pressure downloading recommendations. PMID- 10687651 TI - Clinical efficacy and cost-effectiveness of a new synthetic polymer sheet wound dressing. AB - Stage II and III pressure ulcers present product development and product choice challenges to manufacturers and professional wound care clinicians respectively. We evaluated the clinical performance and cost of use associated with a new synthetic polymer dressing for the management of these wounds. A total of 10 home healthcare patients, each with a Stage II or III pressure ulcer, were enrolled and randomized for wound treatment using either the new polymer hydrogel wound dressing or the leading market hydrocolloid dressing. Dressings were changed on an as needed basis only. The wounds were assessed weekly and parameters recorded using the Bates-Jensen Pressure Sore Status Tool. In addition, the clinical performance of the dressing and treatment costs were evaluated. The overall healing rate for the two groups was similar. However the new polymer hydrogel dressing was found to have a more favorable overall clinical performance evaluation based largely on its more favorable support of autolytic debridement. The new polymeric dressing also had a more favorable cost of use based on the evaluation. We conclude that the new polymer dressing may be a favorable alternative to the leading market hydrocolloid dressing for the treatment of Stage II and III pressure ulcers due to a better clinical performance and the substantially lower treatment costs associated with its use. PMID- 10687653 TI - A moral abomination: food for thought. PMID- 10687652 TI - The effects of UVC irradiation on group A streptococcus in vitro. AB - Streptococcus pyogenes (group A streptococcus--GAS) is a common cause of necrotizing fasciitis (NF)--a severe infection of the subcutaneous soft tissue. The purpose of this study was to determine if the topical therapy ultraviolet light C (UVC) is effective in killing GAS in vitro and to evaluate the most effective treatment parameters for use with UVC therapy. Five replications of GAS at 10(8) organisms/mL were plated. The cultures were treated with a UVC light 1 inch from the surface. Irradiation times were as follows: 0, 2, 3, 4, 5, 15, 30, 45, 60, 90, 120, and 180 seconds. Bacterial cultures were incubated and colony counts performed. A second set of GAS cultures were exposed to UVC for 30, 90, and 120 seconds either once daily (qd) or twice daily (bid). Kill rates were 99.9% for GAS at 4 seconds to 180 seconds. Kill rates of 99.9% were also obtained at 30 seconds and 90 seconds when UVC treatment was given either qd or bid. This data indicates that UVC is bactericidal for GAS at times as short as 4 seconds. In addition, UVC treatment was not effective when administered through thin film dressings. PMID- 10687654 TI - Barriers to informed consent in clinical trials. AB - Clinical trials are a type of scientific study conducted in human beings. They are designed to evaluate the safety and effectiveness of new drugs, procedures, or other means of treating, diagnosing, or preventing diseases. The ethical issues involved in clinical trials are nothing new, but because of the increased access to studies, they are steadily affecting more people. The sheer volume of clinical trials, as well as the number of patients and professionals involved in them, has increased in the past decade. Informed consent is a key element of clinical research. Meaningful informed consent requires that subjects weight the associated risks and benefits of an experimental intervention and then voluntarily give consent. One concern regarding informed consent in research is the confusion between therapeutic intervention and experimental treatment. This confusion could lead to barriers in informed consent. This column addresses this concern and includes a case study to illustrate the misunderstandings that might arise from the patient who is compelled to consent to clinical trials because of his sense of hopelessness from a chronic health condition and the traditional medical intervention he is receiving. PMID- 10687655 TI - Exploring a new path toward global understanding of wound care. PMID- 10687656 TI - The development of a national registration form to measure the prevalence of pressure ulcers in The Netherlands. AB - To gain insight into the prevalence of pressure ulcers in Dutch healthcare institutions, a national registration form to measure the prevalence of pressure ulcers annually in different healthcare settings was developed based on a literature study and responses from a Delphi panel. The reliability and the feasibility of the form devised were tested in a pilot study conducted in a university hospital, a nursing home, and in a home healthcare setting. Interrater reliability of the grading system varied between the institutions from 0.49 to 0.97 (Cohen's Kappa). In the home healthcare setting, interrater reliability was 0.80 (Pearson correlation coefficient) for the total score on the Braden scale. The prevalence rates were 10.1% (n = 368) in the university hospital, 12.7% (n = 1,541) in the home healthcare setting, and 83.6% (n = 122) in the nursing home, although the latter figure seemed to be somewhat exaggerated. The most common lesions were found on the sacrum and below the knee (heel and malleolus). The pilot study concluded that it is possible to collect accurate and reliable data on the scope and severity of pressure ulcers with a uniform instrument in different healthcare settings. PMID- 10687657 TI - The clinical and cost effectiveness of externally applied negative pressure wound therapy in the treatment of wounds in home healthcare Medicare patients. AB - Pressure ulcers, a devastating and costly healthcare problem, often occur in home healthcare settings. We sought to determine if these and other chronic wounds treated at home with negative pressure wound therapy close faster and reduce treatment costs compared to conventional therapies. Records for 1,032 Medicare home healthcare patients with 1,170 wounds that failed to respond to previous interventions--and were subsequently treated with negative pressure wound therapy -were reviewed. Reductions in wound area and volume were compared to rates reported by Ferrell in 1993, and costs were analyzed. Ferrell reported trochanteric and trunk pressure ulcers averaging 4.3 cm2, treated with a low-air loss surface and saline-soaked gauze closed at an average of 0.090 cm2 per day. For comparison to Ferrell's outcomes, we analyzed our Stage III and IV trochanteric and trunk wounds treated with low-air-loss and negative pressure wound therapy. Ours averaged 22.2 cm2 in area and closed at an average of 0.23 cm2 per day. The average 22.2 cm2 wound in our study, treated as described by Ferrell, would take 247 days to heal and cost $23,465. Using negative pressure wound therapy, the wound would heal in 97 days and cost $14,546. The study concluded that negative pressure wound therapy is an efficacious and economical treatment modality for a variety of chronic wounds. PMID- 10687658 TI - Stoma management in a tropical country: colostomy irrigation versus natural evacuation. AB - People with ostomies in Singapore were initially resistant to colostomy irrigation. This study, a prospective crossover study of 26 patients who underwent abdominoperineal resection, compared colostomy irrigation with the natural evacuation method. During the colostomy-irrigation phase of the study, all 26 patients reported an improvement in continence and fewer problems with sleep, sex, and skin complications compared to the natural-evacuation phase. The study also found a reduction in monthly expenses with colostomy irrigation compared to natural evacuation. Patient satisfaction scores were also superior during the colostomy-irrigation phase. This difference in satisfaction scores was less marked in those who were more than 1-year postsurgery than in those who were less than 1-year postsurgery. The difference in satisfaction between colostomy irrigation and natural evacuation scores was statistically significant in the group that was less than 1-year postsurgery, but not in the group that was more than 1-year postsurgery. The study concluded that colostomy irrigation after abdominoperineal resection is superior to natural evacuation in terms of cost and patient satisfaction and should be introduced soon after surgery. PMID- 10687659 TI - Using telemedicine in the treatment of pressure ulcers. AB - Pressure ulcers are dynamic and therefore require frequent assessment and immediate treatment. For many patients who live long distances from rehabilitation hospitals, frequent assessment and immediate treatment are often unavailable. Recent advances during the last two decades have resulted in the development of telemedicine--long-distance delivery of medical education and services to patients. This pilot study reports on a patient enrolled in a telemedicine program during his fifth hospitalization for pressure ulcers in 16 months. Although this is only a single case study, the results suggest the potential efficacy of this new intervention. PMID- 10687660 TI - Entering the 21st century: moving incontinence treatment options to the forefront. PMID- 10687661 TI - A case of ethics and patient rights. PMID- 10687662 TI - Competency in informed consent. AB - Assessing a patient's capacity to make competent decisions concerning her own care is an important clinical skill in healthcare, especially among those who are elderly, chronically ill, and institutionalized. Competency is an important presupposition to autonomous decision making. Assessing competency becomes increasingly critical when the patient's wish is to forego a life-saving procedure. Supporting a patient's choice regardless of the outcome of that decision is an important part of patient advocacy and therefore an important component of patient care. The essence of autonomy is described as it relates to defining meaningful informed consent. This article includes a model for assessing competence. PMID- 10687663 TI - Getting bruised in new places. PMID- 10687664 TI - Health and wound care in Ireland: an American's view. PMID- 10687665 TI - Incontinence and PPS: a new era. AB - Urinary incontinence (UI) is a prevalent and costly problem in nursing homes. Assessing residents with incontinence is necessary to determine the pathophysiologic causes and associated factors that can interfere with self toileting. Nurses can perform this assessment at the bedside. Guideline tools have been developed to assist nursing home staff through the evaluation of UI and intervention. Treatment techniques, specifically behavioral interventions and toileting assistance programs, can be readily incorporated into nursing practice. Most nursing home staffs can easily implement interventions such as bowel and nighttime voiding management and dietary modifications. Nursing home research has demonstrated the effectiveness of toileting assistance programs; however, very little of this research and documented techniques has been used by nursing home staff. Scheduled toileting and bladder training programs can be successfully implemented in nursing home residents. The key to the success of these programs is identifying residents who should be targeted for each specific program. Staff education remains an ongoing issue, as caregivers must be aware of attitudes and beliefs about the aging process and its impact on the genitourinary system in order to provide effective care. Under the Prospective Payment System, nursing homes need to change business as usual and remain abreast of new innovations and research in different behavioral interventions and continence technology. PMID- 10687666 TI - Toileting assistance based on bladder volume. PMID- 10687667 TI - Sleep-disordered breathing as a mechanism for nocturia: preliminary findings. AB - Nocturia is commonly associated with prostate or bladder problems but is also an important symptom of obstructive sleep apnea, a potentially lethal condition. The primary purpose of this study was to test the relationship between symptoms of sleep-disordered breathing and increased nocturnal urine production as described by the Sleep-Disordered Breathing--Nocturia Model. The purpose of the first phase of this three-phase study was to survey community-dwelling older adults (> 55 years) about nocturia and sleep-disturbance symptoms. A random sample of 1,000 older adults, balanced by ethnicity and gender, were surveyed via a mailed questionnaire. The brief questionnaire included characterizing poor sleep quality, obstructive sleep apnea symptoms, nocturia, lower urinary tract symptoms, naps, and self-rated health. The return rate was low (18%, n = 176), but respondents were equally represented by gender and ethnicity across the targeted age groups. Half of the respondents (n = 87) reported > or = 2 voids per night, two-thirds of whom reported nocturia as bothersome. The data showed that African-American women had significant associations between episodes of nocturia and symptoms of obstructive sleep apnea, poor sleep quality, naps, and lower urinary tract symptoms, thus failing to support the notion that nocturia or sleep disordered breathing are prostate or gender related. As expected, subjects (n = 80) who volunteered for the later phases of the study, had significantly more problems. These preliminary data suggest that the relationship between obstructive sleep apnea and nocturia is important because older adults are at higher risk of injury due to falls that may occur while attempting to toilet in the dark. Also, older adults may also be at higher risk of receiving inappropriate urologic treatment if they are not screened for sleep disorders when reporting nocturia along with symptoms of excessive daytime sleepiness and sleep-disordered breathing. Phases II and III of the parent study will include a detailed examination of hormonal, biochemical, and physical variables to further test the proposed Sleep-Disordered Breathing--Nocturia Model. PMID- 10687668 TI - Initial antiarrhythmic drug therapy during resuscitation from sudden cardiac death: a time for a fundamental change in strategy? PMID- 10687669 TI - Pharmacological management of atrial fibrillation: an update. AB - Therapy of atrial fibrillation remains difficult in many patients. There is increasing awareness that antiarrhythmic drug therapy instituted to maintain sinus rhythm after successful cardioversion of atrial fibrillation may pose a substantial risk to the patient. Therefore, results of prospective randomized trials are needed to allow a more evidence-based approach to the treatment of this common arrhythmia. Two recently published studies have shown superiority of amiodarone over conventional antiarrhythmic drugs in maintaining sinus rhythm. The largest such study published today, the Canadian Trial in Atrial Fibrillation (CTAF), has randomized 403 patients to amiodarone or to sotalol or propafenone. At the end of the observation period, amiodarone-treated patients were significantly more likely to remain in sinus rhythm than conventionally treated patients. A number of new antiarrhythmic drugs, mainly class III substances, are currently developed for the treatment of atrial fibrillation or atrial flutter. Ibutilide has recently been released for intravenous administration, attempting pharmacological cardioversion of atrial fibrillation/atrial flutter. It has been evaluated in a number of prospective trials, which showed a higher conversion rate in patients with atrial flutter. Dofetilide is another new compound developed mainly for maintenance of sinus rhythm after restoration of sinus rhythm. It has been evaluated in two prospective, randomized, placebo-controlled trials; moreover, analysis of the DIAMOND trials showed effectiveness of dofetilide in maintaining sinus rhythm in patients with depressed left ventricular function without increased mortality when compared with placebo. Finally, several ongoing studies compare the therapeutic strategy of controlling ventricular rate in atrial fibrillation compared with the strategy of maintaining sinus rhythm. These trials will help to optimize therapy in atrial fibrillation, the most commonly encountered arrhythmia. PMID- 10687670 TI - The importance of considering trial design when interpreting clinical trial results. AB - BACKGROUND: In recent decades, clinical trials have played an increasingly important role in determining how we practice. Trial results proving that a clinical finding poses risk have led to interventions that try to reduce risk. Clinical trials proving that a particular therapy provides better outcome than another therapy have changed the therapies we now use. Unfortunately, the results of clinical trials are too often affected by biases or design issues that may overtly or covertly alter the results or the way they should really be used. In addition, these biases and design and analysis issues are rarely evident in the abstract sections or key figures and tables in the publications reporting the trials, which may be all the busy physician either reads or remembers. METHODS AND MATERIALS: This manuscript discusses the issues involved in optimally understanding clinical trial design and interpretation so that practitioners can better understand how to intelligently read and apply trial results to clinical practice. CONCLUSIONS: Clinical trial results can not be properly applied without consideration of trial design features and intertrial comparisons. PMID- 10687671 TI - Efficacy of atorvastatin compared with simvastatin in patients with hypercholesterolemia. AB - BACKGROUND: Atorvastatin, a new enantiomerically pure synthetic statin, has shown a marked low-density lipoprotein (LDL) cholesterol reduction at doses ranging from 10 to 80 mg/d. This trial was designed to compare the efficacy of atorvastatin 10 mg with simvastatin 10 mg and 20 mg, the latter dose being commonly used in some countries. METHODS AND RESULTS: A parallel group, randomized, PROBE, multicenter study was conducted to compare the efficacy of 10 mg/d atorvastatin with that of 10 mg/d simvastatin and 20 mg/d simvastatin in patients with primary hypercholesterolemia. After a 6-week diet-placebo lead-in period, 272 patients with LDL cholesterol > or = 160 mg/dL and triglycerides < or = 300 mg/dL were randomized to 6 weeks of treatment with atorvastatin 10 mg (109 patients), simvastatin 20 mg (109 patients), or simvastatin 10 mg (54 patients). In the main analysis, which tested the equivalence of atorvastatin 10 mg and simvastatin 20 mg, the mean percent change in LDL cholesterol for atorvastatin 10 mg (-37.0%) was greater than and not equivalent to simvastatin 20 mg (-33.8%). In the secondary analysis, which compared the efficacy of atorvastatin 10 mg with that of simvastatin 10 mg, the mean decrease in LDL cholesterol was significantly greater (P < .001) for atorvastatin 10 mg than for simvastatin 10 mg (-37.0% vs. 28.9%). The two drugs were well tolerated, with an incidence of clinical and biochemical side effects similar among the 3 treatment groups. CONCLUSION: In primary hypercholesterolemia, atorvastatin 10 mg was more effective and nonequivalent to simvastatin 20 mg and significantly more effective than simvastatin 10 mg for reducing LDL cholesterol levels. PMID- 10687672 TI - Acute renal failure after cardiopulmonary bypass: a possible association with drugs of the fibrate group. AB - BACKGROUND: Renal failure is a recognized, but infrequent, complication following cardiac surgery. The causes for this condition are multifactorial, and a major concern is that the occurrence of postoperative acute renal failure is still associated with a high mortality rate. METHODS AND MATERIALS: We report unexpected acute renal failure occurring in 4 patients after uncomplicated cardiac surgery. Each patient was taking a fibric acid derivative at the time of surgery. Renal failure occurred rapidly within 3 days of surgery and was associated with increased concentrations of skeletal muscle-derived creatine kinase (CK). One patient developed myoglobinuria, and another developed a malignant hyperthermia-like syndrome. CONCLUSIONS: These cases show that patients receiving lipid lowering medications could be at higher risk of developing acute renal failure after cardiac surgery. This association merits careful evaluation in large prospective studies and, if proved, would suggest that patients taking either statins or fibrates should discontinue doing so before cardiac surgery. PMID- 10687673 TI - A new model of ventricular plication: a suturing technique to decrease left ventricular dimensions, improve contractility, and attenuate ventricular remodeling after myocardial infarction in the rat heart. AB - BACKGROUND: The Batista procedure (cardio-reduction) is a surgical technique in patients with dilated cardiomyopathy that results in improvement of ventricular function. The purpose of this study was to test a new suturing technique without resection for cardio-reduction of myocardial infarct scars in rats. METHODS AND RESULTS: Myocardial infarction (MI) was induced by occluding the left coronary artery 4 weeks before enrollment. Animals then were randomized to a control (n = 11) or treatment group (n = 11). A pursestring suture was placed within the border zones of the infarcted area and was either tightened (treated) or not (controls). Echocardiography was used to measure left ventricular diameters before, 1 hour, and 6 to 7 weeks after plication. Acutely after plication, end diastolic length (EDL) decreased from 0.70 +/- 0.03 cm to 0.53 +/- 0.02 cm, P < .001; end-systolic length (ESL) decreased from 0.51 +/- 0.03 cm to 0.23 +/- 0.02 cm, P < .001; and fractional shortening (FS) increased from 27.6 +/- 1.5% to 57.6 +/- 2.3%, P < .001, whereas controls were unchanged. In control rats EDL increased from baseline at 0.73 +/- 0.02 cm to 0.82 +/- 0.04 cm at 6 weeks postsurgery, P < .05; ESL increased from 0.54 +/- 0.02 cm to 0.66 +/- 0.04 cm, P < .005; and FS decreased from 26.9 +/- 1.1% to 19.2 +/- 1.2% at 6 weeks, P < .05. In contrast, at 6 weeks in plicated animals, EDL was significantly less than controls at 0.64 +/- 0.02 cm, P < .005; ESL was significantly less than controls at 0.39 +/- 0.03 cm, P < .005, and FS was significantly better than controls at 40.5 +/- 2.2%, P < .005. CONCLUSION: The progressive LV enlargement between 4 and 10 weeks after MI reflects late ventricular remodeling. Plication by suturing infarcted tissue acutely decreases diameters and improves function. At 6 weeks, function remains improved over untreated animals. PMID- 10687674 TI - Beneficial effects of vitamin E treatment in acute myocardial infarction. AB - BACKGROUND: Vitamin E (Vit E), an antioxidant, is considered to prolong survival in patients and animals after myocardial infarction. Because myocardial infarction is associated with arrhythmia and heart dysfunction, this study tested the hypothesis that early treatment with Vit E reduces mortality because of its protective effects against arrhythmia and cardiac dysfunction induced by acute myocardial infarction. METHODS: Rats were randomly divided into 4 groups: sham control, myocardial infarcted, Vit E-treated sham control, and Vit E-treated infarcted animals. Myocardial infarction was induced by ligation of the left anterior descending coronary artery. Treated animals received Vit E (25 mg/kg/d) through a gastric tube beginning 1 hour after the coronary occlusion, whereas control rats received tap water. RESULTS: Electrocardiograms (lead II) at 1, 3, 7, and 21 days after coronary occlusion in the untreated animals showed ST segment elevation, abnormal Q waves, premature ventricular complex (PVC), and QTc prolongation. Conversely, Vit E-treated rats showed attenuated ST-segment changes, fewer abnormal Q waves, and decreased incidence of PVC after coronary occlusion. Total mortality was reduced from 38% to 16%, whereas the infarct size was decreased from 44.2% to 22.3% in infarcted rats treated with Vit E. The depression in left ventricular function as well as elevation of malondialdehyde content and conjugated diene formation in the 21-day infarcted rat hearts were prevented by Vit E treatment. CONCLUSION: These results indicate that Vit E may exert beneficial effects on the heart by reducing oxidative stress in acute myocardial infarction. PMID- 10687676 TI - Transient acoustic wave propagation in rigid porous media: a time-domain approach AB - Wave propagation of acoustic waves in porous media is considered. The medium is assumed to have a rigid frame, so that the propagation takes place in the air which fills the material. The Euler equation and the constitutive relation are generalized to take into account the dispersive nature of these media. It is shown that the connection between the fractional calculus and the behavior of materials with memory allows time-domain wave equations, the coefficients of which are no longer frequency dependent, to be worked out. These equations are suited for direct and inverse scattering problems, and lead to the complete determination of the porous medium parameters. PMID- 10687675 TI - Cardiovascular dysfunction in hypercholesterolemia associated with enhanced formation of AT1-receptor and of eicosanoids. AB - BACKGROUND: In hypercholesterolemia with or without atherosclerosis cardiovascular dysfunction and altered signalling of angiotensin (Ang II), nitric oxide (NO), or prostanoids are intimately related to enhanced oxidant stress and concomitant changes in gene expression. We analyzed cardiac angiotensin receptor (AT1) expression and metabolism of Ang II, eicosanoids, and NO in hypercholesterolemic animals. METHODS: Guinea pigs were fed a 1% cholesterol diet for 8 weeks (Chol). Hemodynamics were analyzed in Langendorff hearts. Spectrophotometric determination of plasma lipids and radioimmunological detection of eicosanoids/cyclic guanosine monophosphate (cGMP). Activities of NO synthase III (NOS-III) or angiotensin converting enzyme (ACE) were determined by enzymatic assays. AT1 receptor density was assessed by radioligand binding assay. NOS-III mRNAs were quantitated by reverse transcription polymerase chain reaction. RESULTS: Hypercholesterolemia was associated with fatty degeneration of the liver and profound myocardial and coronary (e.g., endothelial) dysfunction. In Chol Langendorff hearts we observed significant increases in coronary flow (26.0 +/- 1.0 vs. 17.5 +/- 0.5 mL/min/g tissue) but diminished coronary responses to bradykinin (Bk, 250 ng bolus) or adenosine (Ado, 250 micrograms bolus) (delta CPPBk/Ado: 5 +/- 0.5 vs. 7.2 +/- 1/0.9 +/- 0.1 vs. 1.9 +/- 0.3 cm2 (area under the curve)). AT1 receptor expression was significantly increased in Chol hearts (72 +/- 6.8 vs. 45 +/- 5.6 fmol/mg protein), whereas marked suppression of cardiac activities of ACE (1.96 +/- 0.34 vs. 4.90 +/- 0.20 nmol/min/mg tissue) and of the entire cardiac nitric oxide-cGMP axis (e.g., NOS-III activity: 1.9 +/- 0.4 vs. 3.1 +/- 0.1 pmol/min/mg tissue; NOS-III mRNA: 0.82 +/- 0.16 vs. 1.20 +/- 0.12 arbitrary units; cGMP release: 0.41 +/- 0.02 vs. 0.54 +/- 0.04 pmol/min/g tissue) were shown in Chol. Finally, cardiac release of eicosanoids prostacyclin (PGI2) and thromboxane (TxA2) were significantly enhanced (0.48 +/- 0.05 vs. 0.38 +/- 0.05 and 0.60 +/- 0.10 vs. 0.24 +/- 0.10 ng/min/g tissue, respectively). Enhanced cardiac PGI2 release and suppression of cGMP synthesis in Chol were even more pronounced on stimulation with Bk (38.2 +/- 3.0 vs. 28.2 +/- 2.0 ng/min/g tissue and 1.9 +/- 0.3 vs. 3.0 +/- 0.3 pmol/min/g tissue, respectively). CONCLUSIONS: Altered angiotensin-mediated signal transduction probably related to augmented eicosanoid formation does not compensate for the limited endogenous NO production and for cardiovascular dysfunction in hypercholesterolemic guinea pigs. In this context, changes in redox-sensitive regulation of gene expression (AT1 receptor, NOS-III--caused by enhanced oxidant stress--could play a pivotal role. PMID- 10687677 TI - Backscattering enhancements for tilted solid plastic cylinders in water due to the caustic merging transition: observations and theory AB - Bulk shear and longitudinal waves give rise to important contributions to the scattering of ultrasound by tilted finite plastic and rubber cylinders in water. This occurs in situations where either the shear or longitudinal speed is less than the speed of sound in the surrounding water. At a certain critical tilt angle, large backscattering enhancements are observed for finite cylinders, where the wave vector can reverse direction upon reflection from the cylinder truncation. The scattering process is analogous to the enhancement produced by the merging of rainbow caustics of primary rainbow rays in the scattering of light by long dielectric cylinders, also known as the caustic merging transition [C. M. Mount, D. B. Thiessen, and P. L. Marston, Appl. Opt. 37, 1534-1539 (1998)]. A ray theory was developed to model the backscattering mechanism at the critical tilt angle. It employs the idea of the Bravais effective refractive index, convenient for constructing ray diagrams for the projections of rays in the base plane of the cylinder. There is general agreement between the theory and the experiment down to relatively low ultrasonic frequencies (ka as small as 10). The enhancement is the most significant backscattering contribution for a wide range of tilt angles. PMID- 10687678 TI - Acoustic scattering by a modified Werner method AB - A modified integral Werner method is used to calculate pressure scattered by an axisymmetric body immersed in a perfect and compressible fluid subject to a harmonic acoustic field. This integral representation is built as the sum of a potential of a simple layer and a potential of volume. It is equivalent to the exterior Helmholtz problem with Neumann boundary condition for all real wave numbers of the incident acoustic field. For elastic structure scattering problems, the modified Werner method is coupled with an elastodynamic integral formulation in order to account for the elastic contribution of the displacement field at the fluid/structure interface. The resulting system of integral equations is solved by the collocation method with a quadratic interpolation. The introduction of a weighting factor in the modified Werner method decreases the number of volume elements necessary for a good convergence of results. This approach becomes very competitive when it is compared with other integral methods that are valid for all wave numbers. A numerical comparison with an experiment on a tungsten carbide end-capped cylinder allows a glimpse of the interesting possibilities for using the coupling of the modified Werner method and the integral elastodynamic equation used in this research. PMID- 10687679 TI - New explicit solutions in acoustics of closed spaces on the basis of divergent series AB - A new approach to the acoustics of closed spaces is developed that involves solutions for polygonal shapes in explicit form. It is shown that exact solutions can be constructed for polygonal geometries where all the interior angles are equal to pi/n (n is an integer). It is stated that the set of such polygons consists of the rectangle (known result) and three types of triangles. Some new explicit formulas are obtained for the eigenfrequencies of the triangles. It is demonstrated that the proposed technique also permits an exact representation of the impulse response function for the geometries described. PMID- 10687680 TI - Acoustics of a flanged cylindrical pipe using singular basis functions AB - The problem of acoustic radiation from a cylindrical pipe with an infinite flange has been discussed in a number of papers. The most common approach is to decompose the field inside the pipe over a basis of Bessel functions. A very large number of basis functions is usually required, with a large degree of ripple appearing as an artifact in the solution. In this paper it is shown that a close analysis of the velocity field near the corner yields a new family of functions, which are called "edge functions." Using this set of functions as test functions and applying the moment method on the boundary between the waveguide and free space, a solution is obtained with greatly improved convergence properties and no ripple. PMID- 10687681 TI - Intensity streamlines and vorticity streamlines in three-dimensional sound fields AB - The properties of intensity streamlines and vorticity streamlines are discussed in this paper. It is found that the properties in three-dimensional sound fields are different from the properties in two-dimensional sound fields. The integral behavior of intensity streamlines is that the beginning and the end are attached to a sound source surface or that the beginning is on the sound source surface and the end extends into the infinite. For the vorticity streamlines, the integral behavior is that it is a closed curve or that the beginning and the end are attached to the sound source surface. Three examples are given for intensity and vorticity streamlines. PMID- 10687682 TI - Modified impulse method for the measurement of the frequency response of acoustic filters to weakly nonlinear transient excitations AB - In this paper, a modified impulse method is proposed which allows the determination of the influence of the excitation characteristics on acoustic filter performance. Issues related to nonlinear propagation, namely wave steepening and wave interactions, have been addressed in an approximate way, validated against one-dimensional unsteady nonlinear flow calculations. The results obtained for expansion chambers and extended duct resonators indicate that the amplitude threshold for the onset of nonlinear phenomena is related to the geometry considered. PMID- 10687683 TI - Matched-field processing using measured replica fields AB - An approach for avoiding the problem of environmental uncertainty is tested using data from the TESPEX experiments. Acoustic data basing is an alternative to the difficult task of characterizing the environment by performing direct measurements and solving inverse problems. A source is towed throughout the region of interest to obtain a database of the acoustic field on an array of receivers. With this approach, there is no need to determine environmental parameters or solve the wave equation. Replica fields from an acoustic database are used to perform environmental source tracking [J. Acoust. Soc. Am. 94, 3335 3341 (1993)], which exploits environmental complexity and source motion. PMID- 10687684 TI - Predicting acoustic effects of internal waves from the basic climatology of the world ocean AB - Internal waves of a given strength will produce acoustic effects that vary from water mass to water mass. Presented here is a means of predicting the strength of acoustic fluctuations due to internal waves, given the basic climatology, that is, measurements of depth, temperature, and salinity of an oceanic region. An acoustic fluctuation strength parameter F is defined as the ratio of the fractional potential sound-speed change to the fractional potential-density change. Here F is calculated at three depth levels (275, 550, and 850 m), on a one-degree grid of latitude and longitude, using NODC/OCL's World Ocean Atlas 1994. Representative values of F are presented for 15 upper water masses that range from F = 5 in the North Pacific to F = 34 in the North Atlantic, with a typical value for most of the upper waters being F = 15. Results for two depth levels within 12 intermediate water masses range from F = 7 in the North Pacific to F = 62 in the North Atlantic, with a typical value of F = 20, although there is considerable variation. In general, F exhibits higher values in the Atlantic Basin than in the Indian or Pacific, and has a maximum at 550 m. The main use of F will be the prediction of travel-time fluctuations in acoustic propagation experiments, which will be proportional to the value of F, given a universal strength of internal waves. PMID- 10687685 TI - Long time-base observations of surf noise AB - A year of surf noise observations in the very near shore region of La Jolla Shores beach are presented. Ambient sound levels and surface wave height were recorded for 9 min every hour from July 1997 through June 1998 at a monitoring station located 360 m seaward of the beach in 8-m deep water. Sound segments that were dominated by the noise from breaking surf formed the basis of a correlation analysis between surf noise level and wave height, wave period, wind speed, and mean water depth. The analysis shows that surf noise is primarily determined by wave height, and scales approximately with the wave height squared. The surface wave energy flux onto the beach also scales with wave height squared, leading to the conclusion that the conversion of the mechanical energy of the surface wave field into noise energy is approximately constant. In fact, the ratio of noise energy to surface wave energy flux varies by up to a factor of 3 over the range of energy fluxes considered (100-3000 W per m). PMID- 10687686 TI - Collective oscillations of fresh and salt water bubble plumes AB - Bubble plumes of various void fractions and sizes were produced by varying the flow velocity of a water jet impinging normally on a water surface. The bubbles entrained at the surface were carried downwards by the fluid flow to depths ranging from 33 to 65 cm, and formed roughly cylindrical plumes with diameters ranging from 12 to 27 cm. The acoustic emissions from the plumes were recorded onto digital audio tape using a hydrophone placed outside the cloud at distances ranging from 50 cm to 16.0 m. Closeup video images of the individual bubbles within the plume were also taken in order to gain knowledge of the bubble size distributions. The experiments were performed in both fresh-water and salt-water environments. The fresh-water clouds emitted sounds with a modal structure that was significantly different from that produced by the salt-water clouds. Furthermore, the smaller bubbles present in the salt-water clouds have a fundamental effect on the amplification of turbulence noise, generating sound at significant levels for frequencies up to several hundred Hertz. PMID- 10687687 TI - Ultrasonic absorption in critical binary mixture of perfluoromethylcyclohexane and carbon tetrachloride AB - The results of ultrasonic absorption and velocity measurements for the system perfluoromethylcyclohexane-carbon tetrachloride are presented. In addition, viscosity measurements were made. Ultrasonic absorption at 5, 7, 10, 15, 21, and 25 MHz, above critical temperature Tc, is analyzed using the dynamic scaling theory of Ferrell and Bhattacharjee. The values of alpha/f2 vs f-1.06 show a good agreement with the theory. The experimental values of alpha/alpha c for the binary mixture are compared to the scaling function F(omega*). PMID- 10687688 TI - Comparison between the dispersion curves calculated in complex frequency and the minima of the reflection coefficients for an embedded layer AB - Analytical solutions of Lamb functions for symmetric and antisymmetric elastodynamic modes propagating within a solid layer embedded in an infinite medium are presented. Alternative theoretical analyses of such modes are performed, first in terms of the usual approach of harmonic heterogeneous plane waves (real frequency and complex slowness) and then in terms of transient homogeneous plane waves (complex frequency and real slowness). An example structure of a 0.1-mm-thick "alpha case" (an oxygen-rich phase of titanium that is relatively stiff) plate embedded in titanium is used for the study. A large difference between the usual dispersion curves calculated in real frequency and complex slowness and those calculated in complex frequency and real slowness is shown. Thus the choice between a spatial and a temporal parameter to describe the imaginary part of the guided waves is shown to be significant. The minima and the zeros of the longitudinal and shear plane-wave reflection coefficients are calculated and are compared with the dispersion curves. It is found that they do not match with the dispersion curves for complex slowness, but they do agree quite well with the dispersion curves for complex frequency. This implies that the complex frequency approach is better suited for the comparison of the modal properties with near-field reflection measurements. PMID- 10687689 TI - Measurement of acoustic dispersion using both transmitted and reflected pulses AB - Traditional broadband transmission method for measuring acoustic dispersion requires the measurements of the sound speed in water, the thickness of the specimen, and the phase spectra of two transmitted ultrasound pulses. When the sound speed in the specimen is significantly different from that in water, the overall uncertainty of the dispersion measurement is generally dominated by the uncertainty of the thickness measurement. In this paper, a new water immersion method for measuring dispersion is proposed which eliminates the need for thickness measurement and the associated uncertainty. In addition to recording the two transmitted pulses, the new method requires recording two reflected pulses, one from the front surface and one from the back surface of the specimen. The phase velocity as well as the thickness of the specimen can be determined from the phase spectra of the four pulses. Theoretical analysis and experimental results from three specimens demonstrate the advantages of this new method. PMID- 10687690 TI - The effect of gas loading on the RUS spectra of spheres AB - Resonant Ultrasound Spectroscopy (RUS) of a spherical sample in a pressurizing gas atmosphere was investigated experimentally and theoretically. Measurements were made on a fused silica sphere in He, Ar, and N2 gases up to pressures of 120 bar. The pressure-dependent shift in the resonant frequency, delta f, and the Q factor were measured for the S00, S11, and T02 modes. A theoretical model based on acoustic radiation impedance was used to calculate delta f and the radiation resistance component, Qr, of the Q-factor. Agreement between theory and experiment was good for Qr, but there were discrepancies for delta f. It was found that the theoretical delta f due to gas-loading effects associated with acoustic radiation was very small and consistent with the observed dependence on pressure and gas species for the T02 mode but not for the S00 and S11 modes. We conclude that the T02 mode is the most reliable of these modes to use in measuring third-order elastic constants by RUS. PMID- 10687691 TI - Using phase space diagrams to interpret multiple frequency drive sonoluminescence AB - The recent experimental results of J. Holzfuss, M. Ruggeberg, and R. Mettin [Phys. Rev. Lett. 81, 1961 (1998)] in which a second harmonic drive system was used to generate sonoluminescence (SL) have been analyzed in the context of the dissociation hypothesis (DH) of D. Lohse and S. Hilgenfeldt [J. Chem. Phys. 107, 6986 (1997)]. The second harmonic introduces two more variables that are under experimental control: a phase and an additional pressure term to the acoustic drive pressure. Diffusive equilibrium curves for a fixed gas concentration were calculated as was the Mach criterion. A phase space diagram was constructed to permit the prediction of regions of stable SL, unstable SL, stable non-SL, and unstable non-SL. These were compared to Holzfuss' experimental observations, and excellent quantitative agreement was seen. The results provide further evidence that the underlying assumptions of DH are sound. They also indicate the utility of DH for determining appropriate experimental conditions to achieve SL and for optimizing an experimental system. PMID- 10687692 TI - Use of electrodynamic drivers in thermoacoustic refrigerators AB - Some issues involved in matching electrodynamic drivers to thermoacoustic refrigerators are examined using an equivalent circuit model. Conclusions are that the driver should be chosen to have a large product (Bl)2/(ReRm); the suspension stiffness should be chosen to make the combined impedance of the mechanical and acoustical parts of the system entirely real at the operating frequency; and the piston area should be selected to maximize electroacoustic efficiency, or other desired parameter, by matching the acoustic load to the optimum mechanical load for the particular driver. Alternately, if the piston area is fixed, the operating frequency can be adjusted to make this same match. PMID- 10687693 TI - Acoustical performance of an electrostrictive polymer film loudspeaker AB - A new type of loudspeaker that generates sound by means of the electrostrictive response of a thin polymer film is described. Electrostrictive polymer film (EPF) loudspeakers are constructed with inexpensive, lightweight materials and have a very low profile. The films are typically silicone and are coated with compliant electrodes to allow large film deformations. Acoustical frequency response measurements from 5 x 5 cm (planar dimensions) prototype EPF loudspeakers are presented. Measurements of harmonic distortion are also shown, along with results demonstrating reduced harmonic distortion achieved with square-root wave shaping. Applications of EPF loudspeakers include active noise control and general-purpose flat-panel loudspeakers. PMID- 10687694 TI - Hybrid tool for quickly estimating the radiated acoustic power from a vibrating structure in a multiple-source environment AB - This paper presents a new hybrid method for predicting overestimating and underestimating indicators of the acoustic power radiated by a vibrating surface even in the presence of other surrounding acoustic sources. This method is applicable to plates or low curvature surfaces radiating in open acoustic fields. The method is hybrid in the sense that the vibration field is measured and the parietal pressure field is predicted considering two extreme academic cases "baffled" and "unbaffled." Many simplifications are made and justified in order to save running time. The method is successfully validated in comparison with experimental results on both laboratory and real life structures. This method has led to a quick tool, allowing one to obtain a good approximation of the radiated power in a few minutes. It provides a natural extension of a classical analyzer for vibroacoustics engineering. PMID- 10687695 TI - Rotating machinery dynamics simulation. I. Rigid systems with ball bearing nonlinearities and outer ring ovality under rotating unbalance excitation AB - The radial clearance in rolling bearing systems, required to compensate for dimensional changes associated with thermal expansion of the various parts during operation, may cause dimensional attrition and comprise bearing life, if unloaded operation occurs and balls skid [D. Childs and D. Moyer, ASME J. Eng. Gas Turb. Power 107, 152-159 (1985)]. Also, it can cause jumps in the response to unbalance excitation. These undesirable effects may be eliminated by introducing two or more loops into one of the bearing races so that at least two points of the ring circumference provide a positive zero clearance [D. Childs, Handbook of Rotordynamics, edited by F. Ehrich (McGraw-Hill, NY, 1992)]. The deviation of the outer ring with two loops, known as ovality, is one of the bearing distributed defects. Although this class of imperfections has received much work, none of the available studies has simulated the effect of the outer ring ovality on the dynamic behavior of rotating machinery under rotating unbalance with consideration of ball bearing nonlinearities, shaft elasticity, and speed of rotation. To fill this gap, the equations of motion of a rotor-ball bearing system are formulated using finite-elements (FE) discretization and Lagrange's equations. The analyses are specialized to a rigid-rotor system, by retaining the rigid body modes only in the FE solution. Samples of the results are presented in both time domain and frequency domain for a system with and without outer ring ovality. It is found that with ideal bearings (no ovality), the vibration spectrum is qualitatively and quantitatively the same in both the horizontal and vertical directions. When the ring ovality is introduced, however, the spectrum in both orthogonal planes is no longer similar. And magnitude of the bearing load has increased in the form of repeated random impacts, between balls and rings, in the horizontal direction (direction of maximum clearance) compared to a continuous contact along the vertical direction (direction of positive zero clearance). This underlines the importance of the vibration measuring probe's direction, with respect to the outer ring axes, to capture impact-induced vibrations. Moreover, when the harmonic excitation is increased for a system with ideal bearings, the spectral peaks above forcing frequency have shifted to a higher-frequency region, indicating some sort of a hard spring mechanism inherent in the system. Another observation, is that for the same external excitation, vibration amplitude at forcing frequency in the bearing force spectrum is the same for systems with or without outer ring ovality. PMID- 10687696 TI - Reverberation time and maximum background-noise level for classrooms from a comparative study of speech intelligibility metrics. AB - Speech intelligibility metrics that take into account sound reflections in the room and the background noise have been compared, assuming diffuse sound field. Under this assumption, sound decays exponentially with a decay constant inversely proportional to reverberation time. Analytical formulas were obtained for each speech intelligibility metric providing a common basis for comparison. These formulas were applied to three sizes of rectangular classrooms. The sound source was the human voice without amplification, and background noise was taken into account by a noise-to-signal ratio. Correlations between the metrics and speech intelligibility are presented and applied to the classrooms under study. Relationships between some speech intelligibility metrics were also established. For each noise-to-signal ratio, the value of each speech intelligibility metric is maximized for a specific reverberation time. For quiet classrooms, the reverberation time that maximizes these speech intelligibility metrics is between 0.1 and 0.3 s. Speech intelligibility of 100% is possible with reverberation times up to 0.4-0.5 s and this is the recommended range. The study suggests "ideal" and "acceptable" maximum background-noise level for classrooms of 25 and 20 dB, respectively, below the voice level at 1 m in front of the talker. PMID- 10687697 TI - Broadband control of plate radiation using a piezoelectric, double-amplifier active-skin and structural acoustic sensing AB - The potential of a piezoelectric, double-amplifier active-skin with structural acoustic sensing (SAS) is demonstrated for the reduction of broadband acoustic radiation from a clamped, aluminum plate. The active-skin is a continuous covering of the vibrating portions of the plate with active, independently controllable piezoelectric, double-amplifier elements and is designed to affect control by altering the continuous structural radiation impedance rather than structural vibration. In simulation, acoustic models are sought for the primary and secondary sources that incorporate finite element methods. Simulation indicates that a total radiated power attenuation in excess of 10 dB may be achieved between 250 and 750 Hz with microphone error sensing, while under SAS the radiated power is reduced by nearly 8 dB in the same frequency range. In experiment, the adaptive feed forward filtered-x LMS (least mean square) algorithm, implemented on a Texas Instruments C40 DSP, was used in conjunction with the 6I6O control system. With microphone error sensing, 11.8-dB attenuation was achieved in the overall radiated power between 175 and 600 Hz, while inclusion of SAS resulted in a 7.3-dB overall power reduction in this frequency band. PMID- 10687698 TI - Performance of some sparseness criterion blind deconvolution methods in the presence of noise AB - A comparison of the spareseness (simplicity) norm criterion blind deconvolution methods of Cabrelli and Wiggins is made in order to ascertain relative performance for underwater acoustic transient source signal estimation, especially in the presence of noise. Both methods perform well at high signal-to noise ratios, producing source estimates that are significant improvements over the original received signal for classification purposes. At moderate and lower SNRs, the Cabrelli method tends to generate results that are superior to the Wiggins method. This is especially true for a damped sinusoid transient source, for which the Wiggins method fails completely at lower SNRs, while the Cabrelli method can still produce good source estimates. PMID- 10687699 TI - A macro-mechanical model of the guinea pig cochlea with realistic parameters. AB - The post-mortem transfer function of the cochlea of the guinea pig was compared to the transfer function generated by a model with parameters derived from physical measurements of the guinea pig cochlea. Both the formulation and parameters of the model were carefully chosen to be realistic using evidence from published measurements. The fit between the transfer function of the model and recent mechanical measurements of the passive guinea pig cochlear response was good, with a root mean square ratio of 6.3 dB in amplitude and 0.33 pi rad in phase. The model was used to explore the effect of cochlear partition mode factor and duct geometry upon the mechanical response of the cochlea. Possible inadequacies of the model which could explain the remaining differences between the output of the model and measurements are discussed. PMID- 10687700 TI - Stochastic properties of cat auditory nerve responses to electric and acoustic stimuli and application to intensity discrimination. AB - Statistical properties of electrically stimulated (ES) and acoustically stimulated (AS) auditory nerve fiber responses were assessed in undeafened and short-term deafened cats, and a detection theory approach was used to determine fibers' abilities to signal intensity changes. ES responses differed from AS responses in several ways. Rate-level functions were an order of magnitude steeper, and discharge rate normally saturated at the stimulus pulse rate. Dynamic ranges were typically 1-4 dB for 200 pps signals, as compared with 15-30 dB for AS signals at CF, and they increased with pulse rate without improving threshold or changing absolute rate-level function slopes. For both ES and AS responses, variability of spike counts elicited by repeated trials increased with level in accord with Poisson-process predictions until the discharge rate exceeded 20-40 spikes/s. AS variability continued increasing monotonically at higher discharge rates, but more slowly. In contrast, maximum ES variability was usually attained at 100 spikes/s, and at higher discharge rates variability reached a plateau that was either maintained or decreased slightly until discharge rate approached the stimulus pulse rate. Variability then decreased to zero as each pulse elicited a spike. Increasing pulse rate did not substantially affect variability for rates up to 800 pps; rather, higher pulse rates simply extended the plateau region. Spike count variability was unusually high for some ES fibers. This was traced to response nonstationarities that stemmed from two sources, namely level-dependent fluctuations in excitability that occurred at 1-3 s intervals and, for responses to high-rate, high-intensity signals, fatigue that arose when fibers discharged at their maximum possible rates. Intensity discrimination performance was assessed using spike count as the decision variable in a simulated 2IFC task. Neurometric functions (percent correct versus intensity difference) were obtained at several levels of the standard (I), and the intensity difference (delta I) necessary for 70% correct responses was estimated. AS Weber fractions (10 log delta I/I) averaged +0.2 dB (delta IdB = 3.1 dB) for 50 ms tones at CF. ES Weber fractions averaged -12.8 dB (delta IdB = 0.23 dB) for 50 ms, 200 pps signals, and performance was approximately constant between 100 and 1000 pps. Intensity discrimination by single cells in ES conditions paralleled human psychophysical performance for similar signals. High ES sensitivity to intensity changes arose primarily from steeper rate-level functions and secondarily from reduced spike count variability. PMID- 10687701 TI - Temporal integration in the presence of off-frequency maskers. AB - Temporal integration was measured at a relatively low and a relatively high signal frequency under conditions of off-frequency masking. The masker was typically gated for 300 ms, and the signal was presented 70 ms after masker onset. In experiment 1, the signal frequency was 500 or 2000 Hz. Temporal integration was measured in quiet and in the presence of a masker whose frequency was lower or higher than the signal frequency. In all listening situations, there was less integration at 2000 Hz than at 500 Hz. This effect of frequency was particularly dramatic in the presence of a lower frequency masker, where there was almost no integration at 2000 Hz. Experiment 2 showed that this dramatic effect of frequency cannot be understood in terms of the underlying psychometric functions. Experiment 3 measured temporal integration at 750 and 2000 Hz for a large number of masker-signal frequency separations for both a tonal and a noise masker, and in conditions where the masker was gated or continuous. The results with the gated tonal masker largely confirmed the results of experiment 1. The results with the continuous tonal masker and the gated or continuous noise masker, however, were quite different. In those cases, the amount of temporal integration at both signal frequencies was more or less independent of the masker signal separation; the masked temporal integration was nearly equal to the integration in quiet. Thus based on the conditions evaluated here, off-frequency masked temporal integration differs substantially from integration in quiet only for gated tonal maskers located considerably lower in frequency than the signal. It is unclear how to account for this finding, although it may be related to attentional factors. PMID- 10687702 TI - Psychophysical correlates of contralateral efferent suppression. I. The role of the medial olivocochlear system in "central masking" in nonhuman primates. AB - An extensive physiological literature, including experimental and clinical studies in humans, demonstrates that activation of the medial olivocochlear (MOC) efferent system, by either contralateral sound or electrical stimulation, can produce significant alterations in cochlear function and suggests a role for the MOC system in influencing the auditory behavior of binaural hearing. The present data are from psychophysical studies in nonhuman primates which seek to determine if the noted physiological changes in response to contralateral acoustic stimulation have a perceptual counterpart. Four juvenile Japanese macaques were trained to respond to the presence of 1-s sinusoids, presented to the test ear, in an operant reinforcement paradigm. Thresholds were compared for frequencies ranging from 1.0 to 4.0 kHz in quiet, with thresholds measured when continuous, two octave-band noise, centered on the test tone frequency, was presented in the contralateral ear. Contralateral noise was presented at levels of 10-60 dB above detection threshold for the test-tone frequency. While some variability was evident across subjects, both in the frequency distribution and magnitude (as a function of contralateral noise level), all subjects exhibited an increase, or suppression of thresholds in the presence of contralateral noise. On average, thresholds increased systematically with contralateral noise level, to a peak of 7 dB. In one subject, the threshold increase seen with contralateral noise was significantly reduced when the MOC was surgically sectioned on the floor of the IVth ventricle. The characteristics of the measured shifts in behavioral thresholds, in the presence of contralateral noise reported here, are qualitatively and quantitatively similar to both efferent physiological suppression effects and psychophysical central masking threshold shifts which have been reported previously. These data suggest that at least some aspects of "central masking" are efferent-mediated peripheral processes, and that the term "central masking" may be incorrect. PMID- 10687703 TI - The effects of frequency region and level on the temporal modulation transfer function. AB - Temporal modulation transfer functions (TMTFs) were measured using narrow-band AM and QFM noises with upper spectral edges from 0.6 to 4.8 kHz, and spectrum levels of 10 and 40 dB SPL. The cutoff frequency of the TMTF increases as the upper spectral edge is increased up to 4.8 kHz at low levels, and is constant at higher levels. Sensitivity increases with bandwidth if frequency region is constant. In a second experiment, these results were compared to predictions of a model incorporating peripheral and central limitations to modulation detection. To obtain an estimate of peripheral filtering, frequency selectivity was measured using the notched-noise method, with probe frequencies and levels chosen to parallel those in the first experiment. The TMTF data were then predicted using the model. Predicted cutoff frequencies as a function of the upper spectral edge of the test stimulus were lower than but parallel to those of the subjects at the lower stimulus level. The model predicted only a slight increase in cutoff frequency with level, and thus predicted an increase in cutoff frequency with frequency region at the higher level as well, in contrast to the measured data. These results suggest that there are peripheral and central limitations to temporal resolution, but the psychoacoustically derived auditory filter may be only an indirect measure of peripheral filtering, and/or a more complex model may be needed. PMID- 10687704 TI - On the relative influence of individual harmonics on pitch judgment. AB - Spectral weighting functions were estimated in a pitch-comparison task to assess the relative influence of individual harmonics on listeners' pitch judgment. The stimuli were quasi-harmonic complex tones composed of the first 12 components, with fundamental frequencies ranging from 100 to 800 Hz. On each stimulus presentation the frequency of each harmonic was randomly jittered by a small amount. The perceptual weight for each harmonic was calculated as the correlation coefficient between the binary responses of the listener and the frequency jitters for that harmonic. Although in general the present results conform to previous ones showing the predominant role of several low-ranked harmonics, discrepancies exist in details. Contrary to some previous reports that the dominant harmonics were of fixed harmonic ranks regardless of their frequencies, the current results showed that the dominant harmonics were best described as close to a fixed absolute frequency of 600 Hz. PMID- 10687705 TI - Extracting spectral envelopes: formant frequency matching between sounds on different and modulated fundamental frequencies. AB - The four experiments reported here measure listeners' accuracy and consistency in adjusting a formant frequency of one- or two-formant complex sounds to match the timbre of a target sound. By presenting the target and the adjustable sound on different fundamental frequencies, listeners are prevented from performing the task by comparing the absolute or relative levels of resolved spectral components. Experiment 1 uses two-formant vowellike sounds. When the two sounds have the same F0, the variability of matches (within-subject standard deviation) for either the first or the second formant is around 1%-3%, which is comparable to existing data on formant frequency discrimination thresholds. With a difference in F0, variability increases to around 8% for first-formant matches, but to only about 4% for second-formant matches. Experiment 2 uses sounds with a single formant at 1100 or 1200 Hz with both sounds on either low or high fundamental frequencies. The increase in variability produced by a difference in F0 is greater for high F0's (where the harmonics close to the formant peak are resolved) than it is for low F0's (where they are unresolved). Listeners also showed systematic errors in their mean matches to sounds with different high F0's. The direction of the systematic errors was towards the most intense harmonic. Experiments 3 and 4 showed that introduction of a vibratolike frequency modulation (FM) on F0 reduces the variability of matches, but does not reduce the systematic error. The experiments demonstrate, for the specific frequencies and FM used, that there is a perceptual cost to interpolating a spectral envelope across resolved harmonics. PMID- 10687706 TI - Effectiveness of spatial cues, prosody, and talker characteristics in selective attention. AB - The three experiments reported here compare the effectiveness of natural prosodic and vocal-tract size cues at overcoming spatial cues in selective attention. Listeners heard two simultaneous sentences and decided which of two simultaneous target words came from the attended sentence. Experiment 1 used sentences that had natural differences in pitch and in level caused by a change in the location of the main sentence stress. The sentences' pitch contours were moved apart or together in order to separate out effects due to pitch and those due to other prosodic factors such as intensity. Both pitch and the other prosodic factors had an influence on which target word was reported, but the effects were not strong enough to override the spatial difference produced by an interaural time difference of +/- 91 microseconds. In experiment 2, a large (+/- 15%) difference in apparent vocal-tract size between the speakers of the two sentences had an additional and strong effect, which, in conjunction with the original prosodic differences overrode an interaural time difference of +/- 181 microseconds. Experiment 3 showed that vocal-tract size differences of +/- 4% or less had no detectable effect. Overall, the results show that prosodic and vocal-tract size cues can override spatial cues in determining which target word belongs in an attended sentence. PMID- 10687707 TI - Spectral characterization of jitter, shimmer, and additive noise in synthetically generated voice signals. AB - Alteration of the harmonic structure in voice source spectra, taken over at least two periods of the waveform, may occur due to the presence of fundamental frequency (f0) perturbation, amplitude perturbation, additive noise, or changes within the glottal source signal itself. In order to make accurate inferences regarding glottal-flow dynamics or perceptual evaluations based on spectral measurements taken from the acoustic speech waveform, investigation of the spectral features of each aperiodic component is required. Based on a heuristic development involving a consideration of the partial sum of the Fourier series taken for two periods of a jittered, shimmered, and (additive, random) noise contaminated signal, the corresponding spectral characteristics are hypothesized. Subsequent to this, the Fourier series coefficients are calculated for the two periods in order to test the hypotheses. Definite spectral differences are found for each aperiodic component; based on these findings differential quantitative spectral measurements are suggested. Further supportive evidence is obtained through use of Fourier transform and periodogram-averaged calculations. The analysis is carried out on synthetically generated glottal-pulse waveforms and on radiated speech waveforms. A discussion of the results is given in terms of voice aperiodicity in general and in terms of their implication for future studies involving human voice signals. PMID- 10687708 TI - Quantitative assessment of second language learners' fluency by means of automatic speech recognition technology. AB - To determine whether expert fluency ratings of read speech can be predicted on the basis of automatically calculated temporal measures of speech quality, an experiment was conducted with read speech of 20 native and 60 non-native speakers of Dutch. The speech material was scored for fluency by nine experts and was then analyzed by means of an automatic speech recognizer in terms of quantitative measures such as speech rate, articulation rate, number and length of pauses, number of dysfluencies, mean length of runs, and phonation/time ratio. The results show that expert ratings of fluency in read speech are reliable (Cronbach's alpha varies between 0.90 and 0.96) and that these ratings can be predicted on the basis of quantitative measures: for six automatic measures the magnitude of the correlations with the fluency scores varies between 0.81 and 0.93. Rate of speech appears to be the best predictor: correlations vary between 0.90 and 0.93. Two other important determinants of reading fluency are the rate at which speakers articulate the sounds and the number of pauses they make. Apparently, rate of speech is such a good predictor of perceived fluency because it incorporates these two aspects. PMID- 10687709 TI - The recognition of isolated words and words in sentences: individual variability in the use of sentence context. AB - Estimates of the ability to make use of sentence context in 34 postlingually hearing-impaired (HI) individuals were obtained using formulas developed by Boothroyd and Nittrouer [Boothroyd and Nittrouer, J. Acoust. Sco. Am. 84, 101-114 (1988)] which relate scores for isolated words to words in meaningful sentences. Sentence materials were constructed by concatenating digitized productions of isolated words to ensure physical equivalence among the test items in the two conditions. Isolated words and words in sentences were tested at three levels of intelligibility (targeting 29%, 50%, and 79% correct). Thus, for each subject, three estimates of context ability, or k factors, were obtained. In addition, auditory, visual, and auditory-visual sentence recognition was evaluated using natural productions of sentence materials. Two main questions were addressed: (1) Is context ability constant for speech materials produced with different degrees of clarity? and (2) What are the relations between individual estimates of k and sentence recognition as a function of presentation modality? Results showed that estimates of k were not constant across different levels of intelligibility: k was greater for the more degraded condition relative to conditions of higher word intelligibility. Estimates of k also were influenced strongly by the test order of isolated words and words in sentences. That is, prior exposure to words in sentences improved later recognition of the same words when presented in isolation (and vice versa), even though the 1500 key words comprising the test materials were presented under degraded (filtered) conditions without feedback. The impact of this order effect was to reduce individual estimates of k for subjects exposed to sentence materials first and to increase estimates of k for subjects exposed to isolated words first. Finally, significant relationships were found between individual k scores and sentence recognition scores in all three presentation modalities, suggesting that k is a useful measure of individual differences in the ability to use sentence context. PMID- 10687710 TI - Suprasegmental and segmental timing models in Mandarin Chinese and American English. AB - This paper formalizes and tests two key assumptions of the concept of suprasegmental timing: segmental independence and suprasegmental mediation. Segmental independence holds that the duration of a suprasegmental unit such as a syllable or foot is only minimally dependent on its segments. Suprasegmental mediation states that the duration of a segment is determined by the duration of its suprasegmental unit and its identity, but not directly by the specific prosodic context responsible for suprasegmental unit duration. Both assumptions are made by various versions of the isochrony hypothesis [I. Lehiste, J. Phonetics 5, 253-263 (1977)], and by the syllable timing hypothesis [W. Campbell, Speech Commun. 9, 57-62 (1990)]. The validity of these assumptions was studied using the syllable as suprasegmental unit in American English and Mandarin Chinese. To avoid unnatural timing patterns that might be induced when reading carrier phrase material, meaningful, nonrepetitive sentences were used with a wide range of lengths. Segmental independence was tested by measuring how the average duration of a syllable in a fixed prosodic context depends on its segmental composition. A strong association was found; in many cases the increase in average syllabic duration when one segment was substituted for another (e.g., bin versus pin) was the same as the difference in average duration between the two segments (i.e., [b] versus [p]). Thus, the [i] and [n] were not compressed to make room for the longer [p], which is inconsistent with segmental independence. Syllabic mediation was tested by measuring which locations in a syllable are most strongly affected by various contextual factors, including phrasal position, within-word position, tone, and lexical stress. Systematic differences were found between these factors in terms of the intrasyllabic locus of maximal effect. These and earlier results obtained by van Son and van Santen [R. J. J. H van Son and J. P. H. van Santen, "Modeling the interaction between factors affecting consonant duration," Proceedings Eurospeech-97, 1997, pp. 319-322] showing a three-way interaction between consonantal identity (coronals vs labials), within word position of the syllable, and stress of surrounding vowels, imply that segmental duration cannot be predicted by compressing or elongating segments to fit into a predetermined syllabic time interval. In conclusion, while there is little doubt that suprasegmental units play important predictive and explanatory roles as phonological units, the concept of suprasegmental timing is less promising. PMID- 10687711 TI - Anisotropy of ultrasonic propagation and scattering properties in fresh rat skeletal muscle in vitro. AB - The anisotropy of frequency-dependent backscatter coefficient, attenuation, and speed of sound is assessed in fresh rat skeletal muscle within 5 h post-mortem. Excised rat semimembranosus and soleus muscles are measured in 37 degrees C Tyrode solution, with the muscle fibers at 90 degrees and 45 degrees orientations to the incident sound beam. Reflected and through transmission signals from either a 6- or 10-MHz focused transducer give frequency dependent information in the 4-14 MHz range. The attenuation coefficient in each muscle is consistently a factor of 2.0 +/- 0.4 lower for propagation perpendicular to the fibers than at 45 degrees, whereas speed of sound shows a much milder anisotropy, and is slightly faster for the 90 degrees orientation. The largest anisotropy is seen in the backscatter coefficient, most notably in the semimembranosus where the magnitude at 90 degrees is over an order of magnitude greater than at 45 degrees, with the frequency dependence in both cases giving a power law between 1.5 and 2.0. PMID- 10687713 TI - C-scan imaging in molten zinc by focused ultrasonic waves PMID- 10687712 TI - Spectral cues and perception of the vertical position of targets by the big brown bat, Eptesicus fuscus. AB - Big brown bats (Eptesicus fuscus) were trained to discriminate between vertical angles subtended by paired beads suspended from fishing line. Bats were rewarded for choosing the smaller of the two angles presented. The difference between the angles was changed systematically using a transformed up-down procedure and the bats' ability to detect the difference was measured at different vertical locations. When the beads were centered at +20 degrees (above the horizon), at 0 degree (the horizon), and at -20 degrees (below the horizon), vertical angle acuity (VAA) was maintained between 2.9 degrees and 4.1 degrees. At more extreme vertical positions both bats showed loss of acuity; when the beads were centered around -40 degrees, VAA was 6.7 degrees or 8.3 degrees and at +40, VAA was worse than 21 degrees (the largest difference tested). When the tragi of both ears were bent down and glued to the side of the face, bats showed severe loss of acuity for beads centered at -20 degrees (VAA 18.3 degrees and 20.1 degrees), but maintained their angle acuity for beads centered at +20 degrees (VAA 3.8 degrees and 4.9 degrees). The results are consistent with the spectral cues created by the filtering of the external ear. PMID- 10687714 TI - Coupling of velocity dispersion curves of leaky Lamb waves on a fluid-loaded plate PMID- 10687716 TI - On a numerical truncation approximation algorithm for transfer matrix method PMID- 10687715 TI - Contribution to the hydroacoustic ocean monitoring of the UN Test Ban Treaty; signal classification by an autonomous buoy system PMID- 10687717 TI - Performance limits of the broadband generalized sidelobe cancelling structure in an isotropic noise field PMID- 10687718 TI - Echo suppression in the horizontal and median sagittal planes PMID- 10687719 TI - A speech corpus for multitalker communications research. PMID- 10687720 TI - Directional hearing is only weakly dependent on the rise time of acoustic stimuli. PMID- 10687721 TI - Variability in the characterization of the headphone transfer-function. PMID- 10687722 TI - Acoustical imaging through a multiple scattering medium using a time-reversal mirror. AB - Acoustical imaging is based on the ability to focus an acoustic beam inside the zone of interest. This remains an issue through a high-order multiple scattering medium because the electronic delay lines that enable one to focus through a multiple scattering medium are a priori unknown. Using time-reversal principles, we show that images can be obtained through a very disordered medium. Surprisingly, the images are better than those obtained in a homogeneous medium with a classical imaging device. PMID- 10687723 TI - Acute lymphoblastic leukemia in children. AB - As the overall long-term event-free survival rate in children with acute lymphoblastic leukemia approaches 80%, emphasis is being placed on risk-directed therapy so that patients are neither overtreated nor undertreated. It has become apparent that a risk assignment system based on primary genetic abnormalities is inadequate by itself. For example, leukemias with the MLL-AF4 or BCR-ABL fusion gene are, in fact, heterogeneous diseases. Many require allogeneic hematopoietic stem-cell transplantation; some, if the patient is of favorable age and has a low presenting leukocyte count, can be cured with chemotherapy alone. Measurement of early responses to therapy and extent of minimal residual disease can greatly improve the accuracy of risk assessment. Consideration of the variable effects of therapy on the prognostic significance of specific genetic abnormalities is also important. Therefore, TEL-AML1 fusion confers a favorable prognosis in some protocols of chemotherapy but not in others. Studies to identify genetic polymorphisms with pharmacokinetic and pharmacodynamic significance promise to guide further refinement of treatment strategies. This will allow maximization of anticancer effects without induction of unacceptable toxicity in individual patients. PMID- 10687724 TI - Myelodysplasia. AB - The epidemiology of myelodysplasia, or myelodysplastic syndrome (MDS), is in evolution. As populations are aging and therapies for cancer are improving, the frequency of this disease is increasing. Recent population surveys and case control studies are reviewed. Knowledge of the molecular pathogenesis and pathophysiology of MDS is advancing at a remarkable pace and new information on molecular events is presented. The treatment of MDS is complex and highly individualized. Although many patients are older and may have significant co morbid disease or poor performance status, there are curative options with allogeneic transplantation for selected patients. The recent transplant publications are reviewed. Other investigative treatment approaches, including the use of new chemotherapy agents, growth factor combinations, and antithymocyte globulin appear promising and are reviewed. PMID- 10687725 TI - Update on the biology of chronic lymphocytic leukemia. AB - An understanding of the molecular biology of B-cell chronic lymphocytic leukemia (B-CLL) has led to the appreciation that several different B-cell diseases are represented under this name. Variability in the bcl-2 family of proteins, p53 mutation, or the presence of various chromosomal abnormalities corresponds to variability of the clinical course of disease and response to therapy. Differential expression of cell surface adhesion molecules by B-CLL cells have also been shown to influence clinical outcome, as have the expression of immune regulatory molecules (eg, CD80, CD40R, CD27 and CD79b). Recent work studying immunoglobulin-heavy chain gene rearrangement postulates at least two subsets of B-CLL originating from different stages of B-cell development and following different clinical courses. The knowledge that B-CLL is the final consequence of many different molecular perturbations may allow the development of chemotherapies, immunotherapies, and gene therapies that target the specific molecular defect in a given case of B-CLL. PMID- 10687726 TI - Clinical studies of new "biologic" approaches to therapy of acute myeloid leukemia with monoclonal antibodies and immunoconjugates. AB - Conventional chemotherapeutic treatments of acute leukemias are often associated with life-threatening toxic effects due to a lack of specificity for hematopoietic cells. Monoclonal antibodies and fusion proteins that target antigens on leukemic blasts are being explored for their antileukemic effects and as a means of delivering chemotherapy or radiation directly to malignant cells. This approach might be safer and more effective than current non-specific chemotherapeutic agents. The cell surface antigens CD33 and CD45 are attractive targets. Although CD33 is expressed on acute myelocytic leukemic blast cells from about 90% of patients, normal hematopoietic stem cells lack this antigen, as do essentially all non-hematopoietic tissues. Anti-CD33 antibodies have been engineered to selectively target malignant myeloid and immature normal cells while sparing normal stem cells. Recently, anti-CD33 antibodies have also been used to deliver radiation or a cytotoxic agent directly to leukemic cells. The strategy for using CD45 as a target differs from CD33 in that it is expressed not only by the vast majority of leukemias, but also by normal stem cells. Therefore, 131I-labeled anti-CD45 antibody has been used in combination with conventional preparative regimens for patients receiving marrow transplantation for acute leukemia. Because the receptor for granulocyte-macrophage colony-stimulating factor is expressed by most myeloid leukemias, fusion proteins consisting of granulocyte-macrophage colony-stimulating factor ligand associated with diphtheria toxin have been proposed as a means of delivering a toxic agent directly to leukemic cells. Both unconjugated and conjugated antibodies show significant promise in the treatment of acute myelocytic leukemia. PMID- 10687727 TI - Clinical applicability of the evaluation of minimal residual disease in acute leukemia. AB - Modern chemotherapy can place most patients with acute leukemia into remission. Unfortunately, many of these patients will subsequently relapse. The study of minimal residual disease focuses on the detection of patients destined to relapse despite appearing to be in clinical remission. In addition, the research has demonstrated that some patients appear to co-exist with their leukemia for years, and this suggests a need to re-examine what it means to be "cured" of leukemia. We await trials testing the intervention of "molecular relapse." Data appear to be sufficient to launch such trials in diseases such as pediatric acute lymphoblastic leukemia and the t(1 5;1 7) acute myeloid leukemia. PMID- 10687728 TI - Update on the management of parathyroid tumors. AB - Parathyroid tumors are virtually always benign with an estimated incidence of parathyroid carcinoma causing hyperparathyroidism in only .017% of cases. Virtually all parathyroid neoplasms, including the rare parathyroid carcinoma, are functional and discussion of the management of parathyroid tumors is tantamount to the discussion of primary hyperparathyroidism. The biochemical diagnostic criteria with rare exception is definitive, and the key issue with this functional benign endocrine neoplasm is when to recommend operation and how to ensure optimal results in this curable disease. Patients symptomatic with nephrolithiasis, significant osteoporosis, bone pain, and in some cases constitutional symptoms should undergo a surgical therapy. Also, patients with markedly abnormal laboratory values including a calcium 12.0 mg/dL, or 24-hr urinary calcium >400 mg/day should be treated surgically. The sestamibi nuclear medicine scan has become the best tool available for imaging of abnormal parathyroid glands. This study is positive between 60% and 90% of initial operations and in between 40% and 70% of reoperations. For multi-gland parathyroid disease or hyperplasia, the sensitivity of this test is decreased. Understanding of the ectopic locations of parathyroid adenoma is of utmost importance in the conduct of the parathyroidectomy. For the rare patients with parathyroid carcinoma, aggressive surgical resection with en bloc removal of any adjacent invading structures is the best chance for cure leading to 10-year survival rates of 49%. PMID- 10687729 TI - Advances in the management of adrenal tumors. AB - Adrenal tumors are very common, with the majority being nonhypersecretory and benign and less than 1% being malignant. Most primary adrenal tumors are sporadic, but may be associated with other endocrine and familial disorders, especially pheochromocytoma. All patients with "sporadic" pheochromocytoma should be screened for MEN-2 and Von Hippel-Lindau disease. As in many endocrine tumors, there are no uniform definitive histologic criteria to distinguish malignancy, which is dependent on the clinical behavior of the tumor and is accurately diagnosed in the presence of adjacent organ invasion, recurrence, or distant metastasis. Surgery remains the cornerstone and the treatment of choice for functional and primary malignant adrenal tumors, both for cure and palliation, with low morbidity and mortality. PMID- 10687730 TI - Advances in the diagnosis and management of thyroid neoplasms. AB - Thyroid cancers are still the most common endocrine cancers. They are dominated by well-differentiated carcinomas, including papillary carcinoma, follicular carcinoma, and medullary thyroid carcinoma. Diagnosis is based on fine-needle aspiration cytologic examination. Recently, reverse transcription polymerase chain reaction for the detection of cancer-specific mRNA was shown to be a useful adjunct in both initial diagnosis and detection of recurrent disease. In addition, positron emission tomography has become a valuable tool for staging and surveillance of thyroid cancer. Given the gradual perfection of surgical technique and reduction in complication rates, near-total and total thyroidectomy should be offered to patients with well-differentiated carcinoma. For medullary thyroid carcinoma, near-total and total thyroidectomy with routine central and bilateral functional neck dissection are recommended. So far, no effective treatment exists for anaplastic thyroid carcinoma. PMID- 10687731 TI - Cytogenetics and cancer. AB - Techniques based on fluorescence in situ hybridization (FISH) have bridged the gap between molecular genetics and conventional cytogenetics. Since its introduction in the late 1980s, advanced FISH-based methods have greatly enhanced the cytogenetic analysis of hematopoietic and solid tumors and are rapidly gaining ground in clinical cytogenetic diagnostics. As interest in FISH technologies has grown, it has inspired an era of new FISH-based technologies such as multiplex FISH, spectral karyotyping, and comparative genomic hybridization. In this review, the focus is on the impact of these technologies in the field of cancer genetics. PMID- 10687732 TI - Cytosine methylation and human cancer. AB - Abnormalities of genomic methylation patterns have been attributed a role in carcinogenesis since the early 1980s, when large-scale demethylation of the genome was thought be an early event in multistep colorectal carcinogenesis. In the 1990s, local de novo methylation (with or without global demethylation) at tumor suppressor loci was held to be involved in silencing of tumor suppressor genes. The mechanisms that might mediate methylation and demethylation in carcinogenesis remain obscure, and there are questions as to whether the methylation changes are a cause or consequence of cellular transformation and clonal expansion. It is also important to derive a set of defined criteria by which a tumor suppressor gene can be concluded to have been inactivated by DNA methylation in a manner that contributes to carcinogenesis. PMID- 10687733 TI - Telomeres, telomerase, and cancer: life on the edge of genomic stability. AB - The presence of telomerase activity in most human tumors, but not in many normal somatic tissues, has raised considerable interest in telomerase as a possible anticancer therapy. Recent advances in the cloning and characterization of mammalian telomerase components have paved the way for a more detailed understanding of the role of telomerase and telomere length maintenance in cell proliferation. Here, we summarize the most recent biochemical and genetic evidence suggesting that telomere length maintenance by telomerase is critical to the proliferative ability of some immortalized mammalian cells in culture and in vivo. PMID- 10687734 TI - Aneuploidy and cancer. AB - Numeric aberrations in chromosomes, referred to as aneuploidy, is commonly observed in human cancer. Whether aneuploidy is a cause or consequence of cancer has long been debated. Three lines of evidence now make a compelling case for aneuploidy being a discrete chromosome mutation event that contributes to malignant transformation and progression process. First, precise assay of chromosome aneuploidy in several primary tumors with in situ hybridization and comparative genomic hybridization techniques have revealed that specific chromosome aneusomies correlate with distinct tumor phenotypes. Second, aneuploid tumor cell lines and in vitro transformed rodent cells have been reported to display an elevated rate of chromosome instability, thereby indicating that aneuploidy is a dynamic chromosome mutation event associated with transformation of cells. Third, and most important, a number of mitotic genes regulating chromosome segregation have been found mutated in human cancer cells, implicating such mutations in induction of aneuploidy in tumors. Some of these gene mutations, possibly allowing unequal segregations of chromosomes, also cause tumorigenic transformation of cells in vitro. In this review, the recent publications investigating aneuploidy in human cancers, rate of chromosome instability in aneuploidy tumor cells, and genes implicated in regulating chromosome segregation found mutated in cancer cells are discussed. PMID- 10687735 TI - Is angiogenesis inhibition the Holy Grail of cancer therapy? AB - Over the last several decades, oncology research and cancer treatment have concentrated primarily on the cancer cells. Unfortunately, despite the intensive quest to find new and more effective compounds for chemotherapy, the survival rate of patients has not significantly changed. In 1971 Judah Folkman proposed that a solid tumor cannot grow without inducing angiogenesis. Intensive search for molecules blocking the formation of a tumor-nourishing capillary network has identified several promising agents in experimental models. Some of these angiogenesis inhibitors are in clinical trials, but a clear statement about the efficacy cannot be made yet. What are the current trends in angiogenesis research? What areas should we intensively investigate? Can we find the Holy Grail of cancer therapy in the years to come in order to stop the ineffable suffering of millions of cancer patients? PMID- 10687736 TI - Detection and clinical relevance of micrometastatic cancer cells. AB - The failure to reduce mortality of epithelial cancer patients is probably a result of the early dissemination of cancer cells to secondary sites, which is usually missed by conventional diagnostic procedures used for tumor staging. Individual carcinoma cells present in regional lymph nodes, blood, or distant organs (eg, bone marrow) can be detected by sensitive immunologic or molecular methods. Because the goal of adjuvant therapy is the eradication of occult micrometastatic tumor cells before metastatic disease becomes clinically evident, the early detection of micrometastases could identify those patients who might benefit from adjuvant therapy. In addition, more sensitive methods for detecting such cells should increase knowledge about the biologic mechanisms of metastasis, which might improve the diagnosis and treatment of micrometastatic disease. In this article, the recent developments in sensitive assays used for the detection of individual micrometastatic cancer cells in patients with epithelial tumors are reviewed. PMID- 10687737 TI - Bibliography. Current world literature. Leukemia. PMID- 10687738 TI - Bibliography. Current World Literature. Endocrine tumors. PMID- 10687739 TI - Bibliography. Current World Literature. Cancer biology. PMID- 10687740 TI - Factors affecting disability in patients attending the internal medicine departments of general hospitals. AB - The aim of this study was to investigate the effect of sociodemographic factors, physical factors and mental factors on the physical and social disability of patients attending outpatient clinics of general hospitals. Physical and psychiatric morbidity in 1580 consecutive patients attending the internal medicine department of general hospitals was assessed using a stratified two stage sampling design method. Of the total, 336 patients completed the second stage interview composed of Primary Care Version of Composite International Diagnostic Interview and Groningen Social Disability Schedule to assess sociodemographic, physical and mental factors. In this study, restricted activity days, disability days and Brief Disability Questionnaire were used for the assessment of physical disability, and Groningen Social Disability Schedule was used for social disability. Sociodemographic, physical and mental factors were all related to disability. Among sociodemographic factors, unemployment was associated with physical disability and social disability mildly. Among physical factors, the severity of physical disease was not associated with disability and medically explained somatic symptoms were associated with disability. Furthermore, the mental factor was more strongly associated with physical and social disability. It could be said that the mental factor is more strongly associated with physical and social disability than sociodemographic or physical factors. In addition, even mild mental symptoms not leading to ICD-10 mental disorders affected disability. From the viewpoint of the patients' burden, it is important to assess the mental symptoms as well as physical status in outpatient clinics of internal medicine or primary care. PMID- 10687741 TI - Mental health problems after stroke. AB - We investigated the mental health of 47 subjects (30 men, mean age 63.8+/-7.7; 17 women, mean age 68.9+/-8.7) with the 60-item General Health Questionnaire (GHQ). All the subjects lived at home in a Japanese rural community and were examined from 2 to 3 years after suffering a stroke. Among the subjects, 18 (38.3%) had GHQ scores of 17 or more, which indicated a mental health problem (MHP). The following variables were included in multiple logistic regression analysis: age, sex (men/women), grade of motor paralysis (no/slight/moderate/severe), side of motor paralysis (no/left side/right side/both sides: in analysis, we used dummy variables), paresthesia (no/yes), rehabilitation (need no rehabilitation or participate in rehabilitation/fail to participate in rehabilitation), social support (not needed or sufficient/insufficient) and overall physical recovery (1/2/3: 1 = 67-100, 2 = 34-66, and 3 = 0-33 on a visual analog scale 100 mm long, 100 meaning full recovery). In univariate analysis all variables except age and sex showed statistically significant associations with MHP. In multivariate analysis, only one variable, overall physical recovery', had a statistically independent association with the status of MHP (Odds ratio 4.39, 95% confidence interval 1.46-13.19). The results of logistic regression analysis indicate that the presence of an MHP is more strongly dependent upon subjective assessment about overall physical recovery after stroke than upon physical impairments and the other psychosocial variables. Therefore, in the community setting, the visual analog scale of overall physical recovery is considered to be a simple, valid method for assessing MHP following stroke. PMID- 10687742 TI - Cultural difference in recognition of facial emotional expression: contrast between Japanese and American raters. AB - Using the Japanese and Caucasian Facial Expressions of Emotion (JACFEE) photo set, the relationship between recognition and intensity ratings of universal facial expressions of emotions in 123 Japanese undergraduate students was examined and compared with data reported by American raters. In Japanese raters, although the intensity was rated as high for some of the poses, their correctness scores were poor, suggesting a serious misjudgment of the intended emotions as defined in the JACFEE photo set. Only in Japanese raters were significant relationships between the intensity scores and the percentage correctness scores for sadness detected (r = 0.97, P < 0.0001), but no significant relationship was observed for other emotions. The robust correlation suggests the possibility that Japanese raters might be more responsive to certain emotional expressions when they are fully or intensely expressed. It is proposed that the facial emotional expression paradigm cannot be applied to the psychiatric setting without first refining for cultural differences. PMID- 10687743 TI - Attentional processing of emotional information in obsessive-compulsive disorder. AB - In order to investigate attentional processing of emotional information in obsessive-compulsive disorder (OCD), 14 patients with OCD and 28 normal control (NC) subjects were asked to name the background colors of anxiety-relevant, compulsion-relevant, positive and neutral words (an emotional Stroop color-naming test). The stimulus words were presented subliminally, and supraliminally. The time of subliminal presentation for each subject was determined in advance by the lexical decision task. In the subliminal condition, the delay for anxiety- and compulsion-relevant words, when compared with neutral words, was greater in OCD patients, while no difference was found in NC subjects. In the supraliminal condition, no delay was found for both OCD patients and NC subjects. In other words, OCD patients were more sensitive to threat information when it could not be identified with consciousness. Moreover, the present study compared checking OCD with cleaning OCD in the attentional processing of emotional information. As a result, it was found that checking OCD patients responded more slowly in naming the background color of subliminal emotional words than cleaning OCD patients. The results indicate that OCD patients, especially with checking compulsion, may have a deficit in automatic processing of threat information. PMID- 10687744 TI - Apolipoprotein E phenotypes in healthy normal controls and demented subjects with Alzheimer's disease and vascular dementia in Mie Prefecture of Japan. AB - In order to clarify the association between apolipoprotein E4 (ApoE4) and the pathogenesis of Alzheimer's disease (AD), we analyzed the distribution of the apolipoprotein E (ApoE) phenotypes and the frequency of the apo E alleles epsilon2, epsilon3, and epsilon4 in Japanese healthy controls (n = 1090, an average age of 51.2+/-12.6 years) and demented patients (n=103, mean age of 73.6+/-9.2 years). Demented subjects were divided into three subgroups: early onset AD group (EOAD; n=25, mean age 63.0+/-6.2 years), late-onset AD group (LOAD; n=33, mean age 79.3+/-5.1 years), and vascular dementia group (VD; n=45, mean age 75.3+/-8.0 years). The apolipoprotein E phenotype was determined by isoelectric focusing and immunoblotting. There were no significant differences in the distribution of the apo E phenotypes by gender or age, and the estimated frequencies of epsilon2, epsilon3 and epsilon4 were 0.05, 0.86 and 0.09, respectively, in the normal controls. There was a significant difference in the distribution of the apo E phenotypes between LOAD and elderly controls aged more than 65 years (P<0.0001). The distribution of the apo E phenotypes in EOAD was the same as that in LOAD. The frequency of the epsilon4 allele was significantly higher in LOAD (0.35, P<0.0001) and EOAD (0.28, P<0.0001) than that in the control subjects (0.07), but not in VD (0.12, P=0.1630). The present findings suggest that ApoE4 is related with both EOAD and LOAD, but not with VD, and support the hypothesis that it is a genetic risk factor of AD. PMID- 10687745 TI - Precedents of perceived social support: personality and early life experiences. AB - In order to examine the effects of personality and early life experiences on perceived social support, a total of 97 young Japanese women were investigated. Current interpersonal relationships were measured by an interview modified from Henderson et al.'s Interview Schedule for Social Interaction (ISSI). Personality was measured by Cloninger et al.'s Temperament and Character Inventory. Early life experiences at home and outside of home were also identified in the interview. The number of sources of perceived support was correlated with self directness, while satisfaction with perceived support was correlated with novelty seeking and with low harm avoidance. No early life experiences--early loss of a parent, perceived parenting, childhood abuse experiences, experiences of being bullied and/or other life events--showed significant correlations with the number or satisfaction of supportive people. The quantity and quality of perception of social support differ in their link to personality, and perceived social support may, to some extent, be explainable in terms of personality. PMID- 10687746 TI - Effects of propofol anesthesia on cognitive recovery of patients undergoing electroconvulsive therapy. AB - The effects of different doses of propofol on post-electroconvulsive therapy (ECT) cognitive recovery were evaluated together with the effects on seizure duration and hemodynamic changes during ECT in 15 depressive patients. Propofol attenuated the increase in arterial blood pressure and heart rate in a dose dependent manner compared with thiamylal. Propofol showed a clinically significant anticonvulsant effect during ECT in a dose-dependent manner. There were no significant differences among the four different induction groups in the mean recovery time from anesthesia, however, a low dose of propofol suppressed the early recovery of cognitive function. For early cognitive recovery after ECT, a deep anesthetic level is necessary when the traditional ECT apparatus is used which produces sine curve wave stimuli. PMID- 10687747 TI - Prevalence and symptomatology of comorbid obsessive-compulsive disorder among bulimic patients. AB - This study sought to assess the prevalence and symptomatology of comorbid obsessive-compulsive disorder (OCD) among Japanese subjects who met the DSM-III-R criteria for bulimia nervosa (BN). The Structured Clinical Interview for DSM-III R Patient Version was used to distinguish 26 BN patients with concurrent OCD from 52 BN patients without OCD. Obsessive-compulsive symptoms in BN subjects with concurrent OCD were evaluated using the Japanese version of the Yale-Brown Obsessive-Compulsive Scale. There were no differences in the prevalence of concurrent OCD between BN subjects with and without a lifetime history of anorexia nervosa. Among BN subjects with concurrent OCD, symptoms related to symmetry and order were most frequently identified, followed by contamination and aggressive obsessions, and checking and cleaning/washing compulsions. Bulimia nervosa subjects with concurrent OCD were more likely than subjects without OCD to have more severe mood and core eating disorder psychopathology. Comorbid OCD is a common phenomenon in Japanese bulimics (33%) similar to that suggested in BN subjects in the Western countries. Obsessive-compulsive symptoms related to symmetry and order were most frequently observed in BN subjects with concurrent OCD, which was a similar finding to that reported among restricting anorexic subjects. PMID- 10687748 TI - Autonomic function in the early stage of panic disorder: power spectral analysis of heart rate variability. AB - Previous studies of autonomic nervous system (ANS) function in panic disorder (PD) patients have yielded conflicting results. We speculate that these differences might result from the variety of clinical stages of PD. In order to investigate this, we compared ANS activity in untreated patients in the early stage of PD with control subjects using power spectral analysis of electrocardiogram R-R intervals (PSR-R) in supine rest and during head-up tilt, which was performed according to the maximum entropy method (MEM). It recognizes two main components: high-frequency power (HF), which mainly reflects cardiac parasympathetic activity, and low-frequency power (LF), which reflects both cardiac sympathetic and parasympathetic activity. The patients with PD had significantly higher values for all components of PSR-R only in tilt position total power (TP), LF, and HF than did the control subjects (P<0.01, <0.01, <0.02, respectively). However, the LF/HF ratio which indicated sympathovagal balance did not differ significantly between the two groups in tilt position. Our findings suggest that patients with PD in the early stage of illness have co-activation of sympathetic and parasympathetic nervous systems, which might act to maintain a balance between the two autonomic systems. PMID- 10687749 TI - Delusional disorder: retrospective analysis of 86 Chinese outpatients. AB - Patients who visited the psychiatric outpatient service of Chang Gung Medical Centre, Tao-Yuan, Taiwan during an 8-year period were studied retrospectively. Among the 10,418 outpatients, 86 (0.83%) were diagnosed as having DSM-IV delusional disorder (DD), including 61 (70.9%) with persecutory type, 12 (14.0%) with the mixed type, seven (8.1%) with jealous type, two (2.3%) with somatic type, two (2.1%) with unspecified type, one (1.2%) with erotomanic type, and another one with grandiose type. The ratio of women to men was 0.86. The mean age at onset was 42.4 +/- 15.41 years, with women being older than men. Thirty-seven cases (43.0%) presented with depressive symptoms at their first visit. Subjects were divided into four groups: persecutory type, jealous type, mixed type and others. There were no significant differences between the four groups in terms of gender, age at onset, time-lapse before seeking psychiatric help, the presence of hallucination or the presence of depression. PMID- 10687750 TI - Improvement of subjective work performance among obstructive sleep apnea patients after treatment with continuous positive airway pressure. AB - Obstructive sleep apnea syndrome (OSAS) is a significant problem for some patients presenting with snoring and excessive daytime sleepiness. The 'golden standard' therapy in OSAS is considered to be nasal continuous positive airway pressure (CPAP). The effects of CPAP on work performance in sleep apnoics has not been studied previously. One hundred and fifty-two patients with OSAS participated in an open label study. The patients were diagnosed as suffering from severe OSAS after they underwent overnight polysomnography showing that their apnea indexes were at least 20. The participants answered four questions concerning self-perceived work performance prior to and after using CPAP during 6 months. There were highly statistically significant decreases (P < 0.000001) in work performance difficulties as graded by the patient. The results of this study indicate that CPAP treatment improves subjective work performance in patients suffering from OSAS. PMID- 10687751 TI - Herbal medicine in the treatment of fluvoxamine-induced nausea and dyspepsia. PMID- 10687752 TI - Risperidone-induced anxiety might also develop 'awakening' phenomenon. PMID- 10687753 TI - Symptoms during normal pregnancy: a prospective controlled study. AB - Symptoms of normal pregnancy have received scant attention in the literature and what is reported is largely unsubstantiated. Yet this is an important aspect of antenatal counselling and care which deserves further investigation if symptoms are to be interpreted correctly. Accordingly, we conducted a prospective controlled study of symptoms during normal pregnancy in both primigravidas and multigravidas. A total of 38 symptoms occurred with a significantly different frequency (mainly increased) in the pregnant subjects in the third trimester compared with the controls. Of these a mean of 24.2 symptoms was experienced by each pregnant woman, double that (mean, 11.2) experienced by healthy nonpregnant controls. The 5 symptoms reported most frequently by the pregnant subjects were frequency of micturition, fatigue, pelvic pressure, insomnia and lower backache. However, a wide range of symptoms involving most body systems were reported. This study has established that symptoms of pregnancy are more numerous than mentioned in current obstetric texts and that they can be attributed to the effects of pregnancy. The third trimester is associated with the greatest number of symptoms and there is a marked decline in their number after delivery. PMID- 10687754 TI - Microinvasive adenocarcinoma of the cervix. AB - We evaluated the management of patients with microinvasive adenocarcinoma of the cervix (MIAC), in particular, to determine the place of conservative surgery, and determine if the FIGO classification for MIAC is valid and equivalent to the classification as it applies to microinvasive squamous cancer. A review was undertaken of the database of the Queensland Centre for Gynaecological Cancer (QCGC) from January, 1986 to October, 1998. The records of all patients recorded as having MIAC were retrieved. Microinvasion was defined according to the 1995 FIGO classification as a depth of invasion of no greater than 5 mm and a horizontal dimension of no greater than 7 mm 30 patients were found to have been treated for MIAC. The vast majority (29) were asymptomatic, disease being discovered at the time of routine Papanicolaou smear. There was a 43% incidence of coexisting squamous intraepithelial neoplasia. Multifocal disease was found in 17% of patients and lymph-vascular positivity in 7%. Eighteen patients were treated with radical surgery and 13 with conservative surgery. There were no recurrences over a follow-up interval of 3-116 months. Of the 18 patients treated with radical surgery, none was found to have occult microscopic disease in the parametria or nodal metastases. A total of 27 ovaries were removed, all of which were free of disease. In this small study, MIAC appears to behave in a manner similar to the squamous equivalent. The results provide some justification for the FIGO classification of a microinvasive glandular neoplasm of the cervix. There is some support for a role for conservative surgery in managing this condition, but there is insufficient worldwide experience to make definitive recommendations. PMID- 10687755 TI - A randomized clinical trial comparing oral misoprostol with synthetic oxytocin or syntometrine in the third stage of labour. AB - This is a multicentre, blocked, randomized trial to compare the efficacy of oral misoprostol 400 microg with current injectable uterotonic agents (oxytocin/ Syntometrine) used prophylactically in the third stage of labour. Main outcome measures were blood loss, use of a second uterotonic agent and difference in haemoglobin level from antepartum to postpartum. Data analysis from 863 women showed a statistically significant increase in both the mean blood loss (p < 0.001) and the rate of postpartum haemorrhage > 500 mL, (RR 2.72: 95% C1 1.73 4.27) in the misoprostol group compared to the oxytocin/Syntometrine group. The use of a second uterotonic agent was higher in the misoprostol group (RR 2.89: 95% Cl 2.00-4.18) as well as a greater decrease in postpartum haemoglobin (p = 0.015). Oral misoprostol 400 microg is significantly less effective than the traditional intramuscular uterotonic agents currently used and therefore cannot be considered as a viable option to these agents in the management of the third stage of labour. PMID- 10687756 TI - Survival of patients with epithelial ovarian cancer and the effect of lymphadenectomy in those with stage 3 disease. AB - This study looks at the 10-year survival data in patients with epithelial ovarian carcinoma at the Mercy Hospital for Women, Melbourne. An ovarian cancer database was established at the hospital in 1980, since then 253 patients have been diagnosed with epithelial ovarian cancer. The 5-year survival rates for Stages 1 to 4 are 75%, 55%, 24% and 21% respectively. The 10-year survival rates are 65%, 55%, 16% and 15% respectively. One hundred and thirty patients have been diagnosed with Stage 3 disease. The Gynaecologic Oncology Department at the Mercy Hospital for Women was formalized in 1987. Patients treated for ovarian cancer by the unit since 1987 had a significantly better survival (median 47 months) than patients treated prior to 1987 (median 17 months), p = 0.03. There is much debate as to whether lymphadenectomy in patients with ovarian cancer is of therapeutic value. In this study the patients with Stage 3 disease and who had a lymphadenectomy performed had a better 5-year survival rate of 38% compared to 22% in the group who did not have a lymphadenectomy (p < 0.05). The Stage 3 patients who had negative retroperitoneal nodes had a 10-year survival rate of 51%. There was no difference in survival rate between patients with endometrioid and serous papillary carcinomas. PMID- 10687757 TI - Induction of labour after 1 previous Caesarean section. AB - The aim of this study was, after induction of labour in women with a previous Caesarean section, to compare the outcome in women with a history of a previous vaginal delivery with women who had never delivered vaginally. A retrospective analysis was performed over a 2-year period, in a Dublin teaching hospital. One hundred and three women who had had 1 previous lower segment Caesarean section had labour induced. Particular attention was given to delivery outcome, history of a vaginal delivery, cervical effacement at induction, influence of epidural analgesia, indication for induction and incidence of uterine rupture. The repeat Caesarean section rate after induction was 20.4%. Of the 51 women who had never previously delivered vaginally, the repeat section rate was 37.3% compared with only 3.9% of the 52 women who had previously delivered vaginally (p < 0.01). Fourteen women who had never delivered vaginally had an uneffaced cervix at induction and the repeat Caesarean section rate in this group was 64.3%. The commonest indication for induction was a postdates pregnancy. The use of epidural analgesia was greater in women who had never delivered vaginally. There were 2 cases of uterine scar rupture. Induction of labour following Caesarean section is associated with a significantly higher incidence of repeat Caesarean section in women who have not had a previous vaginal delivery. If the cervix is not effaced at induction, the repeat Caesarean section rate is higher than if the cervix has started to efface. PMID- 10687758 TI - A laparoscopic surgical training and accreditation program up and running. AB - Beginning in March 1997, a training and accreditation program in laparoscopic surgery was established at our hospital according to the training guidelines provided by the Royal Australian and New Zealand College of Obstetricians and Gynaecologists (RANZCOG) (1, 2). Registrars were accredited upon satisfactory completion of supervised surgery at each training level. Consultants seeking clinical privileges in advanced laparoscopic surgery were asked to submit a formal application. By August 1998, 143 minor laparoscopic procedures had been formally assessed resulting in 8 of 9 registrars successfully completing training to level 2 laparoscopy. Thirty-three of 83 (39.8%) advanced laparoscopic procedures were directly supervised and an experienced laparoscopic surgeon was available, if required, for a further 15 procedures. Two consultants undertook additional supervised training before being granted full accreditation for level 3 laparoscopic procedures. Another 2 consultants have been given provisional accreditation for level 3 procedures. Although challenging, implementation of the RANZCOG guidelines on training and accreditation in laparoscopic surgery is quite possible. Greater efforts should be undertaken to establish these guidelines as the 'gold standard' for hospital accreditation committees. PMID- 10687759 TI - Argument for the surgical staging of apparent early endometrial cancer. AB - Cancer of the uterine corpus, commonly referred to as cancer of the endometrium or cancer of the uterus, continues to be the most common pelvic genital malignancy affecting western women (1). In 1998 in the United States 36,100 women were diagnosed with this cancer, and there were 6,300 deaths from this condition in that year. Of concern is that despite a relatively stable incidence over the last decade, the annual number of deaths since 1987 from endometrial cancer has more than doubled (2). Many controversies exist in the management of apparent early endometrial cancer. These include: 1. The role of surgical staging which includes pelvic lymphadenectomy. 2. The role of the subspecialist gynaecological oncologist in primary surgical treatment. 3. Indications for vaginal and external beam radiotherapy. 4. Who is at risk for recurrence? 5. The role of laparoscopic approach to the management of this disease. Despite their importance, these and other issues have not been appropriately addressed by prospective randomized studies. Treatment strategies and algorithms have thus been based upon a combination of clinicopathological studies and uncontrolled reviews. The views expressed in this clinical opinion are those of the Sydney Gynaecologic Oncology Group and reflect our philosophy on the current management of this tumour, supported by recent and appropriate peer reviewed scientific literature. PMID- 10687760 TI - Threatened abortion: prediction of viability based on signs and symptoms. AB - OBJECTIVE: To examine the relationship between signs and symptoms associated with threatened abortion and viability of the pregnancy. DESIGN: A prospective observational study SETTING: A university teaching hospital PARTICIPANTS: One thousand consecutive women presenting with a threatened abortion. INTERVENTION: A structured history and an examination were performed as initial clinical assessment. These were followed by transvaginal sonography to determine the status of the pregnancy. MAIN OUTCOMES: The relationship between individual signs and symptoms and the status of the pregnancy was determined. Logistic regression was performed to determine which signs or symptoms were independent predictors of spontaneous abortion. RESULTS: A history of having passed a tissue mass, the presence of products of conception in the vagina and an open cervix were the only sign or symptom associated with a greater than 90% chance that the pregnancy was non-viable. Logistic regression of signs and symptoms at presentation indicated that maternal age greater than 35 years, a history of passing clots vaginally, vaginal bleeding similar to normal menstruation, increasing vaginal bleeding and discrepancy of 4 or more weeks between the uterine size on examination and that which would have been expected by menstrual dates were significant predictors of nonviable pregnancy. A history of vomiting was predictive of a viable pregnancy. CONCLUSION: The clinical assessment of threatened abortion is unreliable in most cases and should be superseded by ready access to sonographic assessment. PMID- 10687761 TI - Vaginal reconstruction in the fibrotic pelvis. AB - Vaginal reconstruction was performed in 7 patients who had developed vaginal stenosis as a result of extensive pelvic fibrosis following either pelvic irradiation (6 patients) or multiple vaginal procedures (1 patient). Six patients received split thickness skin grafts and 1 patient received an amnion graft in the creation of the neovagina. Five patients achieved a satisfactory final result. All of these patients were sexually active and described adequate sexual function. There was no serious morbidity associated with these procedures. Safe and successful vaginal reconstruction can be performed in a patient with a fibrotic pelvis. PMID- 10687762 TI - More on management of choroid plexus cysts in the mid-trimester fetus. PMID- 10687763 TI - Gestational diabetes mellitus. At what time should the postprandial glucose level be monitored? AB - OBJECTIVE: To compare selected pregnancy outcomes for women with gestational diabetes mellitus (GDM) with management based on testing either 1 hour or 2 hours postprandially according to the ADIPS recommendations. METHODS: Prospective study of consecutive women referred for the medical management of their GDM. Women were allowed to select whether they would test either 1 hour postprandial with a target glucose of < 8.0 mmol/L or 2 hours postprandial with a target glucose of <7.0 mmol/L. Changes to diet and the introduction and adjustment of insulin therapy were designed to maintain postprandial glucose levels below these targets. RESULTS: 166 women elected to test 1 hour postprandial and 101 elected to test 2 hours postprandial. There were no significant demographic differences between these 2 groups. The fetal birthweight, percentage of women requiring insulin and the total daily dose of insulin were similar in both groups. CONCLUSIONS: For women with GDM, monitoring either 1 hour or 2 hours postprandially led to similar outcomes. This would suggest that the ADIPS recommendations are equivalent and therefore women can choose the most convenient time for their postprandial monitoring. PMID- 10687764 TI - Women's knowledge and attitudes about emergency contraception: a survey in a Melbourne women's health clinic. AB - The aim of the study was to determine the level of awareness of emergency contraception in women seeking pregnancy counselling and to investigate their attitudes towards emergency contraception. All women presenting for pregnancy counselling at a Melbourne women's health clinic in October 1997 were invited to complete a questionnaire detailing their contraceptive practices. One hundred and sixty-six questionnaires were distributed and 153 were completed (92% response rate). The majority of this sample population had heard of some form of emergency contraception and knew where to access it. However only 26% knew that emergency contraception should be taken within 72 hours of unprotected intercourse. Although 80% of the sample had heard of emergency contraception (or the morning after pill) only 9% used it in an attempt to prevent this pregnancy. The majority of the women surveyed support the increased availability of emergency contraception by rescheduling it to a non-prescription item and re-packaging as a single treatment. PMID- 10687765 TI - Comparison of epidural and general anaesthesia for elective caesarean delivery according to the effects of apgar scores and acid-base status. AB - The objective of this study was to determine the effects of lumbar epidural anaesthesia on the Apgar score and acid-base status of the newborn. Umbilical artery blood gases were obtained in 85 singleton, term, uncomplicated pregnancies delivered by elective Caesarean section. The umbilical artery blood pH, PaCO2, PaO2 and HCO3 values and Apgar scores (1 and 5 minutes) were compared between lumbar epidural and general anaesthesia groups. General anaesthesia was used in 45 (52.9%) women and lumbar epidural anaesthesia in 40 (47.1%). Only 2 of the newborns exposed to epidural anaesthesia had umbilical artery blood pH values 7.19 or less. The mean umbilical artery blood pH was found to be significantly lower in the newborns exposed to lumbar epidural anaesthesia (p = 0.011). None of the newborns in the 2 groups were severely depressed (Apgar scores less than 4). The mean umbilical artery blood PaCO2, PaO2 and HCO3 values did not show any significant difference between the groups. In conclusion, lumbar epidural anaesthesia is associated with lower umbilical artery blood pH values, occasionally with severe fetal acidaemia. PMID- 10687766 TI - Oral methotrexate in the management of refractory interstitial cystitis. AB - To establish the safety and efficacy of low-dose oral methotrexate in treating refractory interstitial cystitis, 9 women who fulfilled internationally accepted criteria for the diagnosis of interstitial cystitis were enrolled in a prospective study. All had proven unresponsive to conventional treatment modalities. Assessment by pain score and frequency volume charts was performed pretreatment and up to 6 months during therapy. No significant adverse side effects were noted. At the end of follow-up, 4 women had noted a subjective improvement in bladder pain and wished to continue on methotrexate, 4 women noted little change and 1 woman reported a worsening of symptoms. Overall there was a significant reduction in pain score (p = 0.047) posttreatment. However, there was no significant difference in urinary frequency per 24 hours (p = 0.40), maximum voided volume (p = 0.089) or mean voided volume (p = 0.59). Methotrexate significantly improved bladder pain in women with interstitial cystitis, although no significant change was found in voiding pattern. PMID- 10687767 TI - A retrospective review of perioperative complications in 360 patients who had Burch colposuspension. AB - This retrospective study reviews intraoperative and early complications of Burch colposuspension of 360 patients. Ten patients had massive haemorrhage and 8 of them had a blood transfusion. Three patients had a haematoma. Bladder injuries were noticed in 10 patients, 3 of whom were diagnosed postoperatively. One patient had unilateral ureteral kinking. Urinary retention occurred in 20 patients for more than 10 days and 2 required catheterization for 26 and 32 days respectively. Eighteen patients had a wound infection and 4 had a wound abscess. Twenty nine patients had a urinary infection. Urinary tract injury, haemorrhage and blood transfusion were significantly more common in women having secondary surgery than those having primary surgery. Deep venous thrombosis was diagnosed in 3 patients who had a Burch colposuspension with concomitant abdominal hysterectomy. Knowledge of possible risks and complications of Burch colposuspension may help plan a better preoperative work-up of patients and may minimize the intraoperative complications and increase surgical success and patient satisfaction. PMID- 10687768 TI - Chronic pain as the main presenting symptom of depression following hysterectomy in old age. PMID- 10687769 TI - Glucose intolerance and other cardiovascular risk factors in chinese women with a history of gestational diabetes mellitus. AB - Women with a history of gestational diabetes (GDM) are at increased risk of developing diabetes compared with other women. There are few data on associations between GDM and cardiovascular risk factors. Between 1988 and 1995, 801 Chinese women with a history of GDM were recruited for a 75g oral glucose tolerance test (OGTT) and assessment of various cardiovascular risk factors, namely obesity, hypertension and dyslipidaemia, 6 weeks after delivery at the Diabetes Centre of the Prince of Wales Hospital. Another 431 women with no past history of diabetes or GDM recruited in a diabetes prevalence study were used as control subjects. After adjustment for age, body mass index and smoking, the prevalence of glucose intolerance remained higher in women with a history of GDM when compared to normal controls. The relative risks of obesity, hypertension, dyslipidaemia, diabetes and impaired glucose tolerance in women with a history of GDM comparing to normal subjects were, respectively, 2.4, 7.5, 2.4, 8.1 and 5.0. After excluding those with abnormal glucose tolerance, subjects with a history of GDM still had more adverse cardiovascular risk factors, including higher blood pressure, glycaemic and lipid parameters, than control subjects (after adjustment for age, body mass index and smoking). In conclusion, compared with normal subjects, Chinese women with a history of GDM had an 8-fold increased risk of having diabetes based on their OGTTs performed 6 weeks postdelivery. These women also have increased rates of other cardiovascular risk factors including obesity, high blood pressure and dyslipidaemia. PMID- 10687770 TI - Estimation of oxidative products of nitric oxide (nitrates, nitrites) in preeclampsia. AB - Oxidative products of nitric oxide, serum nitrates and nitrites were estimated in 50 primigravidas with preeclampsia and in 50 gestation and age-matched normotensive primigravidas. Thirty three (66%) of these women had mild preeclampsia and 17 (34%) had severe preeclampsia. Serum nitrate and nitrite levels were significantly higher in preeclamptic women (nitrates - 15 +/- 1.17; nitrites - 11.82 +/- 1.16 micromol/L) than in the normotensive pregnant women (nitrates 11.82 +/- 1.16; nitrites - 5.08 +/- 0.47 micromol/L, p < 0.001). In preeclamptic women, serum nitrate and nitrite levels correlated with the severity of the disease (mild preeclampsia nitrate - 14.46 +/- 1.98; nitrite 6.21 +/- 0.84 micromol/L, severe preeclampsia nitrate - 16.65 +/- 3.64; Nitrite - 6.87 +/- 1.56 micromol/L). In preeclampsia there was significant positive correlation between nitrate and nitrite levels and diastolic blood pressure and proteinuria. PMID- 10687771 TI - Transvaginal chorionic villus sampling-an alternative approach. PMID- 10687772 TI - Choriocarcinoma developing after prolonged molar surveillance. PMID- 10687773 TI - A cervical ectopic pregnancy managed by medical treatment and angiographic embolization. AB - Medical treatment in the form of systemic methotrexate with or without local methotrexate/potassium chloride is effective for early cervical pregnancy. It should be the treatment of choice in suitable cases. Haemorrhagic complications can be effectively managed by angiographic embolization. A case report illustrating these points is presented. PMID- 10687774 TI - Prenatal diagnosis and management of sacrococcygeal teratoma. AB - Five fetuses, each with a sacrococcygeal teratoma (SCT) were delivered at the Royal Women's Hospital while 2 fetuses, each with a SCT were delivered at Monash Medical Centre in 1998. The number of cases reported in this series is higher than expected but it most likely occurred due to chance. The diagnosis was made prenatally in all cases. Three of the SCT were entirely external while the remaining 4 were external with intrapelvic extension. Rapid growth of the SCT occurred in 3 fetuses. This was associated with polyhydramnios in 2 fetuses. No fetus developed nonimmune hydrops. Six infants were liveborn (perinatal mortality rate of 14%), 3 of whom were delivered prior to 37 weeks' gestation. Two infants were delivered by classical Caesarean section. The remaining 4 infants were delivered by lower uterine segment Caesarean section. There was 1 perinatal death. This stillborn infant was delivered vaginally. The 6 surgical resections were performed between the 4th and 10th postnatal days. Histological examination confirmed the diagnosis of benign SCT in each. One infant developed a recurrence at 2 months of age and required chemotherapy. PMID- 10687775 TI - Pilonidal disease of the pubic region. PMID- 10687776 TI - Pregnancy and cirrhosis of the liver. AB - Pregnancy in patients with cirrhosis of the liver is uncommon. We reviewed 9 pregnancies in 7 patients with cirrhosis. One patient conceived within 1 month of diagnosis and in another the disease was diagnosed during the index pregnancy. Four patients has associated portal hypertension and 1 of them conceived after lienorenal shunt. Complications associated with these pregnancies were jaundice (1) jaundice plus ascites (2) and gastrointestinal bleeding (1). In 2 patients endoscopic sclerotherapy was done during the index pregnancy. The incidence of preterm delivery was 50% (4 of 8) and the majority (75%) occurred in pregnancies where associated complications were present. There was 1 maternal death in the postpartum period due to fulminant hepatic failure. PMID- 10687777 TI - Twin pregnancy in a uterus didelphys, with unilateral placental abruption and onset of labour. AB - A dizygotic twin pregnancy with a fetus in each side of a uterus didelphys is described. An antepartum haemorrhage at 26 weeks' gestation, with subsequent onset of contractions in the right-sided uterus, precipitated delivery by Caesarean section. PMID- 10687778 TI - A new indication for breech extraction. PMID- 10687779 TI - Complete hydatidiform mole with coexisting normal fetus-report of two cases. PMID- 10687780 TI - Endometrial ossification in infertile patients-report of 3 cases. PMID- 10687781 TI - Pelvic sepsis complicating embolization of a uterine fibroid. PMID- 10687782 TI - Conservative management of a case of placenta praevia percreta. AB - Pregnancies complicated by placenta praevia and a history of caesarean section are associated with increased risk of placenta percreta (1). Placenta praevia percreta sometimes involves the bladder or other pelvic organ, invasion leading to genital bleeding or haematuria (2, 3). Bladder injury or uncontrollable profuse haemorrhage occasionally occurs in such patients during surgery. Examination of placental invasion is necessary as this clinical condition is severe. Treatment of placental myometrium invasion is required to prevent uncontrollable profuse haemorrhage during surgery. We present a multiparous patient who was diagnosed prenatally with placenta praevia percreta using magnetic resonance imaging (MRI) and who was treated conservatively with a good prognosis. PMID- 10687783 TI - Pyomyoma: a case report. PMID- 10687784 TI - A severe case of post-antibiotic clostridium difficile colitis. PMID- 10687785 TI - Birth asphyxia, cerebral palsy and litigation. PMID- 10687786 TI - Caesarean section delivery on registrar experience in vaginal breech delivery. PMID- 10687787 TI - Embolic occlusion for the treatment of uterine myomas. PMID- 10687788 TI - Advances in ultrasound biomicroscopy. AB - The visualisation of living tissues at microscopic resolution is attracting attention in several fields. In medicine, the goals are to image healthy and diseased tissue with the aim of providing information previously only available from biopsy samples. In basic biology, the goal may be to image biological models of human disease or to conduct longitudinal studies of small-animal development. High-frequency ultrasonic imaging (ultrasound biomicroscopy) offers unique advantages for these applications. In this paper, the development of ultrasound biomicroscopy is reviewed. Aspects of transducer development, systems design and tissue properties are presented to provide a foundation for medical and biological applications. The majority of applications appear to be developing in the 40-60-MHz frequency range, where resolution on the order of 50 microm can be achieved. Doppler processing in this frequency range is beginning to emerge and some examples of current achievements will be highlighted. The current state of the art is reviewed for medical applications in ophthalmology, intravascular ultrasound, dermatology, and cartilage imaging. Ultrasound biomicroscopic studies of mouse embryonic development and tumour biology are presented. Speculation on the continuing evolution of ultrasound biomicroscopy will be discussed. PMID- 10687789 TI - Significant changes in transrectal ultrasonic measurements of the prostate in relation to the degree of rectal wall distension. AB - The present study was conducted to reveal the possible changes in transrectal ultrasonic measurements of the prostate in relation to the degree of rectal wall distension. When analyzed together for 51 men, all measurements but area changed statistically significantly as the rectal wall was distended by a balloon covering a probe. Ultrasonic measurements concerning the prostatic shape changed more remarkably than those concerning its size. More importantly, changes in ultrasonic measurements were much more remarkable in patients with a healthy prostate than in those with an advanced BPH. These results suggest that possible changes in prostatic shape with the rectal wall distension has to be taken into account when evaluating transrectal prostatic ultrasonograms in terms of changes in shape, especially in patients with a healthy prostate. This is also the case when the diagnosis of BPH is made based on the change in shape, such as presumed circle area ratio, which is a parameter representing the roundness of the horizontal sonogram of the prostate. PMID- 10687790 TI - Prevalence and risk factors of low quantitative ultrasound values of calcaneus in Korean elderly women. AB - Quantitative ultrasound (QUS) of bone is a new radiation-free, low-cost method that measures both bone mass and bone quality. This study was performed to establish the normative data of QUS for Korean women and to determine the prevalence and risk factors of low quantitative ultrasound values in a Korean elderly population. We studied 238 healthy women aged 20-29 years working at a hospital, and 552 women over 50 years of age living in six villages of Chung-Up district, a rural area of South Korea, using QUS measurement of bone. Broadband ultrasound attenuation and speed of sound were measured at the calcaneus, and an index combining these factors (stiffness index) was calculated. T-score was calculated from the data of young normal subjects. Of the 552 elderly women, 34.2% had T-scores between -1.0 and -2.5, and 11.8% had T-scores below -2.5. The prevalence of low quantitative ultrasound values increased with older age, longer duration following menopause, lower body mass index, younger age at menopause and smoking. In multiple logistic regression analysis, age (odds ratio = 1.40 per 5 years, P < 0.05), duration following menopause (odds ratio = 1.35 per 5 years, P < 0.05) and body mass index (odds ratio = 0.78 per quartile, P < 0.05) were independently associated with low quantitative ultrasound values. These results suggested that quantitative ultrasound measurement of the calcaneus could be a useful tool for epidemiological surveys of bone mass. PMID- 10687791 TI - Colour Doppler ultrasound: a new index improves the diagnosis of renal artery stenosis. AB - The description of a new index, the renal-segmental ratio (RSR), and the comparison of its performance with other conventional Doppler parameters for the detection of renal artery stenosis (RAS). A total of 96 renal units were studied with angiography and colour Doppler ultrasound (US) independently. The Doppler parameters applied were: 1. renal artery peak systolic velocity (RE-PSV), 2. renal-aortic ratio (RAR), 3. early systolic acceleration (ESA), and 4. renal segmental ratio (RSR). The angiographic study was used as the "gold standard" for the identification of > or = 50% RAS. The results indicate that RSR (sensitivity: 93.33%, specificity: 89.47%) and RE-PSV (sensitivity: 83.33%, specificity: 89.47%) were the best criteria for RAS diagnosis (p values <0.05). The results show that colour Doppler US is a reliable diagnostic modality for RAS diagnosis. The new index (RSR) improves the effectiveness of the method. PMID- 10687792 TI - Reactive pulmonary artery vasoconstriction in pulmonary consolidation evaluated by color Doppler ultrasonography. AB - A total of 122 patients with pulmonary consolidation on chest radiographs underwent color Doppler ultrasonography to evaluate hemodynamic change in regional pulmonary artery in pulmonary consolidation due to various causes. The diseases underlying pulmonary consolidation included 66 simple pneumonia, 37 obstructive pneumonia, 13 tumor consolidation and 6 pulmonary infarctions. Blood flow signals in consolidation were detected by color-flow mapping. The degree of reactive vasoconstriction was evaluated from analysis of the spectral waveform of the blood flow in the segmental pulmonary artery by several vessel resistance indicating Doppler ultrasound (US) indices, including pulsativity index (PI), resistive index (RI), and acceleration time (AT). The results showed that reactive vasoconstriction was most marked in obstructive pneumonia, followed by simple pneumonia, and least in tumor consolidation (p < 0.001, Kruskal-Wallis test and p < 0.001, Dunn's test for comparison of PI, RI and AT values between different groups of pulmonary consolidation). No flow was detected in pulmonary infarction. We conclude that color Doppler US is a useful tool for evaluating reactive vasoconstriction in pulmonary consolidation. The different degrees of reactive vasoconstriction may be helpful in exploring the possible etiology of pulmonary consolidation. PMID- 10687793 TI - Vertebral arteries and neck rotation: Doppler velocimeter and duplex results compared. AB - The purpose of this study was to test the validity of Doppler ultrasound (US) velocimeter examination of vertebral arteries during contralateral (to the opposite side) cervical rotation. Vertebral arteries from 20 subjects were insonated using a bidirectional Doppler velocimeter at the suboccipital portal (standard technique) and C2 transverse process level (new technique) during contralateral cervical rotation. The results, regarding persistence or major reduction in Doppler signals, were then compared with those from a colour-flow duplex US scanner using the same procedure. There was complete agreement between the combined suboccipital and C2 velocimeter results and those from the duplex scanner (k = 1.00 at p = 0.01): both sensitivity (n = 5) and specificity (n = 34) were 100%. This study provides evidence to support the validity of bidirectional Doppler velocimeter examination, by an experienced examiner, for the purpose of assessing the effects of contralateral rotation on vertebral artery blood flow. PMID- 10687794 TI - High-frequency color flow imaging of the microcirculation. AB - The extension of ultrasound (US) color flow imaging (CFI) techniques to high frequencies (> 20 MHz) has the potential to provide valuable noninvasive tools for scientific and clinical investigations of blood flow in the microcirculation. We describe the development of a slow-scan CFI system operating in the 20-100-MHz range that has been optimized to image the microcirculation. The apparatus has incorporated elements of a previously reported pulsed-wave Doppler system and is capable of operating in either CFI or pulsed-wave mode. The performance of the CFI system was evaluated at a center frequency of 50 MHz using two PVDF transducers with -6-dB beam widths of 43 and 60 microm. The -6 dB-axial resolutions were estimated to be 66 and 72 microm, respectively. In vivo validation experiments conducted using the murine ear model demonstrated the detection of flow in vessels down to 15-20 microm in diameter with flow velocities on the order of mm per s. Further experiments examining experimental murine tumors confirmed the successful detection of flow in the tumor microcirculation. PMID- 10687795 TI - Classification of arterial plaque by spectral analysis of in vitro radio frequency intravascular ultrasound data. AB - To test whether radio-frequency analysis of coronary plaques predicts the histological classification, r.f. data were collected using a 30 MHz intravascular ultrasound scanner. Two hundred ninety-nine regions-of-interest from eight postmortem coronary arteries were selected and identified by histology as falling into one of seven different tissue types. These are loose fibrous tissue (n = 78), moderate fibrous tissue (n = 27), dense fibrous tissue (n = 33), microcalcification (n = 14), calcified plaque (n = 55), lipid/fibrous mixture (n = 51) and homogeneous areas of lipid pool (n = 29). On the basis of a previous study, four spectral parameters were calculated for each of the regions-of interest: maximum power (dB), mean power (dB), spectral slope (dB/MHz) over the bandwidth 18-35 MHz and the intercept of the spectral slope with the 0 Hz axis (dB). A minimum-distance classifier using the Mahalanobis (1948) distance was applied to the data. Following resubstitution of the training data into the classifier, the total correctly classified was 54%. The data were reclassified using three broader tissue groups: (1) calcified plaque, (2) lipid pool and (3) a mixed fibrous category, incorporating loose fibrous tissue, moderate fibrous tissue, dense fibrous tissue, lipid/fibrous mixture and microcalcification. The total correctly classified was 86%. Using "leave-one-out" cross-validation, the classification rates were 48% for seven tissue subgroups and 83% for three broader categories of tissue type. PMID- 10687796 TI - Fundamental studies on contrast images from different-sized microbubbles: analytical and experimental studies. AB - Microbubbles are very useful as ultrasound (US) contrast agents because of their excellent scattering properties. Because microbubbles of different sizes can be used for this purpose, the contrast images produced by different-sized microbubbles are studied in this paper. The contrast images from microbubbles of average sizes 35.5 microm and 2.1 microm were investigated experimentally. Although a low concentration of microbubbles produces contrast-enhanced images without artefacts, an excess of microbubbles results in distorted images. From experimental observation, the distortion of an image caused by microbubbles of average size 35.5 microm was mainly due to multiple scattering, and that by 2.1 microm microbubbles was due to the acoustic shadowing effect. With the use of the tissue-mimicking phantoms of known acoustical properties, the brightness of the contrast images from the microbubble suspension was calculated. The calculated and experimental results of the contrast images produced from microbubbles of average size 35.5 microm were closer to each other when there was no image distortion. When image distortion caused by multiple-scattering occurred, the experimental pixel brightness was higher. For smaller microbubbles of average size 2.1 microm, calculated results of free microbubbles showed a weaker contrast effect than the experimental results. By taking the effect of the coatings of microbubbles into consideration, the calculated brightness of contrast images became much closer to the experimental one. PMID- 10687797 TI - Ultrasonic characterization of the nonlinear properties of contrast microbubbles. AB - The nonlinear properties of microbubble contrast agents have been used to create contrast-specific imaging modalities such as harmonic imaging and subharmonic imaging. Thus, a better understanding of the nonlinear performance of contrast microbubbles may enhance the diagnostic capabilities of medical ultrasound (US) imaging. The first and second harmonic, the 1/2 order subharmonic and the 3/2 order ultraharmonic components in spectra of scattered signals from Optison microbubbles insonified at 2 and 4 MHz have been investigated using an in vitro laboratory pulse-echo system. The development of these signal components over time is quite different for 2-MHz insonification compared to 4-MHz insonification. Scattered subharmonic and ultraharmonic signals are much more time-dependent than first and second harmonic echoes. The dependence of the first and second harmonic, subharmonic and ultraharmonic components on acoustic pressure for 2-MHz insonification is similar to that for 4-MHz insonification. The first and second harmonic components increase linearly with acoustic pressure (in double logarithmic scales) and the subharmonic and ultraharmonic amplitudes undergo rapid growths in the intermediate acoustic pressure range and much slower increases at both lower and higher acoustic pressures. PMID- 10687798 TI - Evaluation of an experimental system for the in vitro assessment of ultrasonic contrast agents. AB - The ultrasonic properties of microbubble contrast agents need to be fully understood if reproducible images and quantitative results are to be produced. Additional aspects of the physical and chemical environment into which the contrast agents are introduced also need to be taken into account, and their effect on contrast agent performance evaluated. A setup that provides an accurate and reproducible data-acquisition system is presented and evaluated in this paper. The linear range of this system is assessed, as well as its accuracy and precision. A new approach to the investigation of contrast agents, based on normalised backscatter, is discussed. Also, a common technique of degassing, widely used in other areas, is described and evaluated to determine its appropriateness to contrast agent studies. PMID- 10687799 TI - In vitro flow quantification with contrast power Doppler imaging. AB - To evaluate the effectiveness of contrast harmonic (power Doppler imaging) as an ultrasonic modality to quantify flow, an in vitro model of perfusion was studied using Optison, a second-generation ultrasound (US) contrast agent. The in vitro model was made of two dialysis cartridges placed parallel and allowed absolute and relative flow quantification on both tube (entry lines) and tissue (cartridges) simulations. Video intensity curves were generated using intermittent harmonic power Doppler imaging after bolus injection of contrast. Correlation between flow and different parameters extracted from time-intensity curves and previously defined as indicators of flow was established for both tissue and entry lines, for flow rates ranging from 0 to 400 mL/min. Single compartment equations were also tested on the model. A good correlation for the tissue model was observed between absolute flow and onset time (O), time to maximal enhancement (TME), peak intensity (P), area under the curve (AUC), and maximal ascending slope (S) parameters, with a r = 0.94, 0.94, 0.91, 0.92 and 0.92, respectively. The correlation for O, TME, P and AUC parameters was r = 0.86, 0.90, 0.78 and 0.82, respectively for entry lines. The correlation for tissue model and entry line was slightly improved when comparing flow ratios with peak ratios (P1/P2) and slope ratios (S1/S2) (r = 0.95 and 0.94). Flow calculation using the gradient-relationship method also showed a good correlation (r = 0.88) with the experimental flow. The results obtained indicated that absolute and relative quantification of flow using PDI is feasible in tube and tissue models. Several clinical applications, namely in myocardial, hepatic and renal artery studies, could be derived from these results. PMID- 10687800 TI - A real-time two-dimensional pulsed-wave Doppler system. AB - An experimental system was developed to acquire and visualise in real-time two dimensional (2-D) velocity maps. Data acquisition is performed by using a modified commercial echograph based on a 5-MHz, 128-element linear-array transducer with electronic focussing and beam steering. Additional electronics were integrated into the echograph to implement a 2-D Doppler system capable of measuring the velocity component on the scanning plane. Suitable axial and lateral scanning methods were studied to obtain Doppler measurements over a scanning area. A colour image of the estimated velocity field is presented in real time on a personal computer using different visualisation techniques. The system performance was tested experimentally both in vitro and in vivo on a human carotid artery. PMID- 10687801 TI - An interactive tool to visualize three-dimensional ultrasound data. AB - Three-dimensional ultrasound can provide images that are easily understood by people who are not specialists in ultrasonography. However, current visualization methods do not perform very well on 3-D ultrasound data. Apart from some specific cases (obstetrics, cardiology), 3-D ultrasound images have not yet demonstrated major benefits from a clinical point of view. In this article, we introduce an interactive method that allows the user easily to produce 3-D images for each ultrasound examination. It is a two-step method. First, the user roughly segments the data by drawing three boundary curves in perpendicular planes. A ray-casting algorithm then automatically retrieves the details of the objects. Because it can be used routinely, this tool should help to evaluate the potential of 3-D ultrasonography. PMID- 10687802 TI - Effects of phase aberration on high frame rate imaging. AB - A high frame-rate (HFR) imaging method (about 3750 frames/s for imaging of biological soft tissues at a depth of 200 mm) has been developed recently with limited diffraction beams. This method uses the fast Fourier transform (FFT) and inverse fast Fourier transform (IFFT) to construct images, and can be implemented with simple and inexpensive hardware, compared to the conventional delay-and-sum method where a digital beam former is usually used. In this paper, phase aberration effects are studied for both the high frame rate and the conventional methods by adding random phase shifts to echo signals obtained from an experiment. In the study, two broadband linear arrays were used to construct images of an ATS 539 tissue-equivalent phantom that has a frequency-dependent attenuation of about 0.5 dB/MHz/cm. The first array has 48 elements, a central frequency of 2.25 MHz, an aperture of 18.288 mm, and a width of 12.192 mm in elevation. The second has 64 elements, a central frequency of 2.5 MHz, and a dimension of 38.4 mm x 10 mm. The-6dB pulse-echo bandwidth of both arrays is about 40% of their center frequencies. Radiofrequency (RF) signals were digitized at 20 mega samples/s at a 12-bit resolution to construct images. Results show that phase aberration has about the same effect on both methods in terms of image resolution and contrast, although the high frame-rate method can be implemented with a simpler system. PMID- 10687803 TI - New piezoelectric transducers for therapeutic ultrasound. AB - Therapeutic ultrasound (US) has been of increasing interest during the past few years. However, the development of this technique depends on the availability of high-performance transducers. These transducers have to be optimised for focusing and steering high-power ultrasonic energy within the target volume. Recently developed high-power 1-3 piezocomposite materials bring to therapeutic US the exceptional electroacoustical properties of piezocomposite technology: these are high efficiency, large bandwidth, predictable beam pattern, more flexibility in terms of shaping and definition of sampling in annular arrays, linear arrays or matrix arrays. The construction and evaluation of several prototypes illustrates the benefit of this new approach that opens the way to further progress in therapeutic US. PMID- 10687804 TI - Effects of ultrasound and 1,25-dihydroxyvitamin D3 on growth factor secretion in co-cultures of osteoblasts and endothelial cells. AB - It has been shown that low-intensity pulsed ultrasound (US) accelerates fracture healing in animal models and in clinical studies. However, the mechanism by which US accelerates fracture healing remains unclear. Systemic factors and several growth factors, such as platelet-derived growth factor (PDGF), are thought to be involved in the process of fracture healing. In the present study, we examined the effects of US and 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on growth factor secretion in a co-culture system of human osteoblastic cells (SaOS-2) and endothelial cells (HUVEC). US was applied to cultured cells for 20 min daily for four consecutive days. US treatment increased the PDGF-AB level in the conditioned media. 1,25-(OH)2D3 (1 x 10(-8) M) also enhanced PDGF-AB secretion. The secretion of PDGF-AB was synergistically increased by the combination of US and 1,25-(OH)2D3. These results suggest that the stimulation of growth factor secretion from cells by US and 1,25-(OH)2D3 treatment may be involved in the acceleration of fracture healing. PMID- 10687805 TI - Nurses' and doctors' attitudes towards suicidal behaviour in young people. AB - This paper presents an exploratory study performed to identify the attitudes towards suicidal behaviour in young people, amongst nurses (and nursing lecturers), and doctors working in in-patient medical and mental health care settings. The Suicide Opinion Questionnaire (SOQ) was administered to 59 participants. Responses were scored using eight clinical scales, and tested by using a Kruskal-Wallis one way analysis of variance. An Independent Sample t-test was used to analyse gender differences. Qualitative interviews were conducted in a sample of respondents. SOQ findings revealed no overall significant differences in the relevant groups of nurses and doctors, with the exception of gender and the clinical scale relating to a 'Cry for Help'. The focused interviews generated five categories relating to suicidal behaviour and young people. Nurses and doctors working in these areas possess a range of influential perceptions of suicidal behaviour and need to be considered in the contexts of care and treatment of young people. PMID- 10687806 TI - Student nurses' lived experience of preceptorship. Part 1--in relation to learning. AB - In this first part of a research project a phenomenological approach was applied to understanding student nurses' experience of preceptorship. The lived experience of seventeen student nurses learning within a preceptor-preceptee relationship on hospital wards formed the basis of the study. Tape-recorded interviews were conducted, transcribed and analysed. The phenomenological hermeneutic analysis revealed three themes of meaning central to the lived experience of learning. These were (a) directing learning; (b) learning in practical action and (c) feeling in learning. The themes included six sub-themes with internal variations. The results indicate that learning, as a phenomenon, could be understood as being in different modes of learning, including directing learning, learning in practical actions and feeling in learning. Each theme was also found to be inherent in the others in an ongoing changeable process. The findings were compared with Aristotle's five modes of learning and revealed that the student nurses' learning embraced scientific knowledge, technical skills, practical wisdom and limited intuition. The study may provide nurse educators with some insight into student nurses' learning by being in real-life situations and performing nursing actions within a preceptor-preceptee relationship. PMID- 10687807 TI - Student nurses' lived experience of preceptorship. Part 2--the preceptor preceptee relationship. AB - Student nurses' experience of preceptorship was the focus of this second part of a phenomenological study. The aim was to illuminate student nurses' lived experience of the preceptor-preceptee relationship on hospital wards. A phenomenological-hermeneutic analysis was made of tape-recorded interviews with seventeen student nurses. The interpretation process culminated in four themes, namely: (a) creating space for learning; (b) providing concrete illustrations; (c) exercising control and (d) seeking reflection. Each theme included sub-themes with internal variations. The theme 'creating space for learning' was understood as basic in relation to the other themes and as the foundation of student learning and preceptoring. The ongoing process of preceptoring meant that the preceptors acted as role models, were with the students and also exercised control. Control was directed both to patient safety and to student learning. The students' 'seeking reflection' included attempts to find peace and quiet either by themselves or with the preceptor. The reflection together with the preceptor facilitated the students' transformation of knowledge, from the specific situation to a general knowledge and increased the value of learning. PMID- 10687808 TI - Caregiver burden and health promotion. AB - Few of the studies describing caregiver stress and burden have focused on the effects of caregiving on the ability of the caregiver to attend to his or her own health needs. Therefore, the major purpose of this study was to investigate whether the perception of burden is related to the health-promoting behaviors of caregivers of the elderly. One hundred twenty-one predominantly female caregivers, mean age 61.1 years, S.D. = 13.4, completed questionnaires measuring demographic and health-related factors, the Objective and Subjective Burden Scales, and the Health-Promoting Lifestyle Profile. Findings suggest that those perceiving lower subjective burden practice more health-promoting behaviors than those with higher subjective burden scores, confirming Pender's contention that situational factors, such as caregiver burden, may affect health promotion. PMID- 10687809 TI - Qualitative research to identify racialist discourse: towards equity in nursing curricula. AB - Professional curriculum planning is beginning to address issues of equity. The authors report on findings from a research initiative to begin to integrate antiracism into an undergraduate curriculum. Theory and methods of Essed, Fanon, Frankenberg, Hall, van Dijk and Woodward are synthesized for interpreting racialist discourse. The findings support the principle of normalizing accountability for discourse practices which construct whiteness and otherness in their representations. Essentialist discourse practices are implicated in the perpetuation of racism, ableism, heterosexism, ageism, etc. Hence, the ideal of equity is expanded to include the enactment of non-essentialist discourse. The logic is revealed as either/or; either equity or dominance through normalized perpetuation of essential categories assigning negative value to others constructing difference, marginalization, problematization, exclusion and containment. The confused, middle or neutral position is one of condoning racism and other forms of dominance. PMID- 10687810 TI - The alcohol use disorders identification test (AUDIT): validation of an instrument for enhancing nursing practice in Hong Kong. AB - This paper describes the psychometric analysis of the alcohol use disorders identification test (AUDIT) after it was modified for use in Hong Kong and administered to examine the patterns of hazardous and harmful drinking. The modified version of AUDIT was an 18-item instrument in which 10 items were completely adopted from the original version and 8 items were added to improve its cultural sensitivity. It was translated into Chinese and back translation was undertaken to confirm the equivalence of the Chinese and English versions. Following a pilot study the instrument was administered to 450 subjects who were recruited from two acute general hospitals, a University Health Clinic and three community health centres. The content validity was judged as adequate by a panel of five international and local experts and the instrument achieved a high reliability coefficient of 0.99 during a test-retest procedure conducted with 20 subjects. Factor analysis was performed on the responses obtained from 450 subjects which supported the construct validity of the 18-item instrument. The modified instrument had a consistently high internal consistency reliability (Cronbach's alpha=0.96-0.97) when tested in the different settings. It was found that a higher percentage of respondents from the hospitals (14.5%) drank at a hazardous or harmful levels compared to those from the community (6.2%) or the University (5.3%). The AUDIT proved a reliable and valid measure with potential applications in Chinese cultures. Early intervention and identification of 'at risk' drinking by the AUDIT is supported as a strategy to be implemented by nurses in primary and secondary health care settings in Hong Kong, where there are indications of increasing alcohol overuse. PMID- 10687811 TI - Developing a model of collaborative research: the complexities and challenges of implementation. AB - While the benefits of collaborative research have been well documented, fewer publications exist regarding the complex, problematic issues involved in these undertakings. This paper offers an integrated collaborative research model to depict the complexities and challenges of initiation and implementation of a 2 1/2 year joint research project between a community hospital and a university school of nursing in Southern Ontario, Canada. A sampling of the experiences of the researchers are analyzed to clarify the dynamic and often competing issues and interactions involved in encouraging hospital-wide research involvement during periods of organizational growth and change. The model reflects the simultaneous interaction of organizational, change and collaborative processes while maintaining the rigor of the research (RCT), and ensuring minimal disruption to the service agency. Quantitative outcomes of this collaboration are presented through an analysis of participant involvement on multiple organizational levels. Recommendations for future collaborative research, including design and methodological issues and collaborative and change strategies are offered. The complexity of balancing the necessary trade-offs required of successful collaborative research are highlighted and will be useful to those considering and planning future collaborations. PMID- 10687812 TI - Creating and maintaining 'optimism' in cancer care communication. AB - This study investigates nurse-patient communication in the cancer care context. Interviews with nurses and patients about their communication experiences and audio-recorded nurse-patient conversations were collected and analysed. A theme of 'optimism' largely manifesting as 'constructive realism' was one of four features identified by the qualitative analysis. The health professional has traditionally been viewed as the party with the power and control over conversation progression and topics. In particular, the superficial, positive and chatty nature of nurse-patient interaction has often been attributed to a lack of nurses' communication skills training. This research indicates that both patient and nurse are active in its construction and argues that the optimistic cheerful nature of nurse-patient interaction may be better viewed as a jointly produced institutional feature of cancer care. This paper illustrates and examines some of the ways this outcome was created and maintained by participants and discusses the implications of this. PMID- 10687814 TI - Complexity of the simplest phylogenetic estimation problem. AB - The maximum-likelihood (ML) solution to a simple phylogenetic estimation problem is obtained analytically The problem is estimation of the rooted tree for three species using binary characters with a symmetrical rate of substitution under the molecular clock. ML estimates of branch lengths and log-likelihood scores are obtained analytically for each of the three rooted binary trees. Estimation of the tree topology is equivalent to partitioning the sample space (space of possible data outcomes) into subspaces, within each of which one of the three binary trees is the ML tree. Distance-based least squares and parsimony-like methods produce essentially the same estimate of the tree topology, although differences exist among methods even under this simple model. This seems to be the simplest case, but has many of the conceptual and statistical complexities involved in phylogeny estimation. The solution to this real phylogeny estimation problem will be useful for studying the problem of significance evaluation. PMID- 10687813 TI - The xylem of rice (Oryza sativa) is colonized by Azorhizobium caulinodans. AB - Following inoculation with Azorhizobium caulinodans ORS571 (pXLGD4), lateral root development of rice and colonization of lateral root cracks by bacteria were shown to be stimulated by the flavonoid naringenin. Rice seedlings growing aseptically in the presence of naringenin were inoculated with ORS571 (pXLGD4), carrying the lacZ reporter gene. By microscopic analysis of sections of inoculated rice roots, it has been demonstrated that the xylem of rice roots can be colonized by Azorhizobium caulinodans. We discuss whether this colonization of the xylem of rice roots by azorhizobia could provide a suitable niche for endophytic nitrogen fixation. PMID- 10687815 TI - Gene flow in the European corn borer Ostrinia nubilalis: implications for the sustainability of transgenic insecticidal maize. AB - Strategies proposed for delaying resistance to Bacillus thuringiensis toxins expressed by transgenic maize require intense gene flow between individuals that grew on transgenic and on normal (referred to as refuges) plants. To investigate gene flow in the European corn borer, Ostrinia nubilalis (Hubner), the genetic variability at 29 sampled sites from France was studied by comparing allozyme frequencies at six polymorphic loci. Almost no deviations from Hardy-Weinberg expectations occurred, and a high stability of allelic distribution was found among samples collected in the same site over two or three different generations, indicating a high stability of the genetic structure over time. The overall genetic differentiation was low at the region and whole country level, suggesting a high and homogeneous gene flow. These results are discussed in relation to the sustainability of transgenic insecticidal maize. PMID- 10687817 TI - Non-random fitness variation in two populations of Darwin's finches. AB - Darwinian fitness of an individual is measured by the number of recruits it contributes to the next generation. We studied variation in fitness among members of three cohorts of two species of Darwin's finches living on the Galipagos island of Daphne Major: the medium ground finch (Geospiza fortis) and cactus finch (Geospiza scandens). Individuals of both species live for up to 16 years. Variation in fitness was neither random nor heritable. Non-randomness arises as a result of a few individuals living for an exceptionally long time and breeding many times. For each cohort, the number of recruits per breeder is strongly predicted by the number of fledglings per breeder. In turn, the number of fledglings is strongly predicted by longevity of the breeder. These results suggest that the most important determinant of fitness is the ability of an individual to survive to breed in many years. Morphological traits affect this ability. Although morphological traits are heritable they do not change unidirectionally because they are selected in opposite directions, and in different combinations, under fluctuating environmental conditions. Non-random fitness variation in fluctuating populations implies much smaller genetically effective sizes than breeding population sizes. PMID- 10687816 TI - Mild environmental stress elicits mutations affecting fitness in Chlamydomonas. AB - Cultures of Chlamydomonas were exposed to a range of relatively mild stresses for a period of 24 h. These stresses comprised high and low temperatures, osmotic stress, low pH, starvation and toxic stress. They were then allowed to recuperate for around ten vegetative generations under near-optimal conditions in unmodified minimal medium. Fitness was then assayed as the rate of division of isolated cells on agar. We found that there was a strong tendency for stressed cultures to have lower mean fitness and greater standardized variance in fitness than the negative controls which had been cultured throughout in unmodified minimal medium. The same tendency was shown, as expected, by positive controls which received mutagenic doses of ultraviolet irradiation. We concluded that the most reasonable interpretation of these observations is that mild stress increases the genomic rate of mutation. This appears to be the first time that this phenomenon has been noticed in eukaryotes. The response might be adaptive because lineages in which higher mutation rates are elicited by stress can be favourably selected through the production of a few mutants which are fortuitously well adapted to the stressful environment. Other interpretations are not excluded, however. Regardless of the mechanism involved, the elevation of mutation rates under stress will affect the rate of evolutionary response to environmental change and also the maintenance of sexuality. PMID- 10687818 TI - Dispersal and extinction in fragmented landscapes. AB - Evolutionary and population dynamics models suggest that the migration rate will affect the probability of survival in fragmented landscapes. Using data for butterfly species in the fragmented British landscape and in immediately adjoining areas of the European continent, this paper shows that species of intermediate mobility have declined most, followed by those of low mobility, whereas high-mobility species are generally surviving well. Compared to the more sedentary species, species of intermediate mobility require relatively large areas where they breed at slightly lower local densities. Intermediate mobility species have probably fared badly through a combination of metapopulation (extinction and colonization) dynamics and the mortality of migrating individuals which fail to find new habitats in fragmented landscapes. Habitat fragmentation is likely to result in the non-random extinction of populations and species characterized by different levels of dispersal, although the details are likely to depend on the taxa, habitats and regions considered. PMID- 10687819 TI - Adaptive significance of male parental care in a monogamous mammal. AB - Paternal behaviour presumably evolved because male care of young was critical for offspring survival. We report field evidence indicating that paternal behaviour enhances offspring survival in a monogamous mammal, the biparental California mouse, Peromyscus californicus. Male removal resulted in lower offspring survival in father-absent than in father-present families. New males took up residence with widowed females, but usually after females had stopped lactating, suggesting that the importance of the father is not primarily protection against infanticidal intruders but rather direct care of young. PMID- 10687820 TI - The importance of stable schooling: do familiar sticklebacks stick together? AB - Preferences for rejoining shoals composed of familiar individuals have recently been documented in a variety of small, shallow-water fish species. Such preferences are assumed to be adaptive, since familiar groups have improved anti predator defences and more stable dominance hierarchies. However, the design of these studies may have created conditions that elevate preferences for familiar individuals. Furthermore, in natural habitats, where significant opportunities for inter-shoal transfer may exist, it is unclear whether shoals stay together long enough for such preferences to develop. Here we present the results of a laboratory study examining whether prior familiarity influences the subsequent shoal composition of sticklebacks (Gasterosteus aculeatus) allowed to re-assort freely in a large arena tank. We show that fish from different familiarity groups associate with familiar conspecifics significantly more than predicted by a model of random assortment, suggesting that even when there is ample opportunity for inter-group transfer, shoal composition can remain stable. We discuss the phenomena that may lead to the formation of familiar groups in natural habitats. In addition, we suggest that familiarity benefits may reduce the relative value of transferring to otherwise more attractive (e.g. larger or more phenotypically matched) groups, and thereby stabilize shoal structure. PMID- 10687821 TI - Foraging rate versus sociality in the starling Sturnus vulgaris. AB - It is well established that social conditions often modify foraging behaviour, but the theoretical interpretation of the changes produced is not straightforward. Changes may be due to alterations of the foraging currency (the mathematical expression that behaviour maximizes) and/or of the available resources. An example of the latter is when both solitary and social foragers maximize rates of gain over time, but competition alters the behaviour required to achieve this, as assumed by ideal free distribution models. Here we examine this problem using captive starlings Sturnus vulgaris. Subjects had access to two depleting patches that replenished whenever the alternative patch was visited. The theoretical rate-maximizing policy was the same across all treatments, and consisted of alternating between patches following a pattern that could be predicted using the marginal value theorem (MVT). There were three treatments that differed in the contents of an aviary adjacent to one of the two patches (called the 'social' patch). In the control treatment, the aviary was empty, in the social condition it contained a group of starlings, and in a non-specific stimulus control it contained a group of zebra finches. In the control condition both patches were used equally and behaviour was well predicted by the MVT. In the social condition, starlings foraged more slowly in the social than in the solitary patch. Further, foraging in the solitary patch was faster and in the social patch slower in the social condition than in the control condition. Although these changes are incompatible with overall rate maximization (gain rate decreased by about 24% by self-imposed changes), if the self-generated gain functions were used the MVT was a good predictor of patch exploitation under all conditions. We discuss the complexities of nesting optimal foraging models in more comprehensive theoretical accounts of behaviour integrating functional and mechanistic perspectives. PMID- 10687822 TI - The evolution of song repertoires and immune defence in birds. AB - Song repertoires (the number of different song types sung by a male) in birds provide males with an advantage in sexual selection because females prefer males with large repertoires, and females may benefit because offspring sired by preferred males have high viability. Furthermore, males with large repertoires suffer less from malarial parasites, indicating that a large repertoire may reflect health status. We hypothesize that sexual selection may cause a coevolutionary increase in parasite virulence and host immune defence because sexual selection increases the risk of multiple infections that select for high virulence. Alternatively, a female mate preference for healthy males will affect the coevolutionary dynamics of host-parasite interactions by selecting for increased virulence and hence high investment by hosts in immune function. In a comparative study of birds, repertoire size and relative size of the spleen, which is an important immune defence organ, were strongly, positively correlated accounting for almost half of the variance. This finding suggests that host parasite interactions have played an important role in the evolution of song repertoires in birds. PMID- 10687823 TI - Predator avoidance and immune defence: costs and trade-offs in snails. AB - Organisms are often confronted by both predators and pathogens. Defending against such widely divergent enemies requires more than one type of defence. Multiple defences, however, raise the possibility of trade-offs among defences. We tested for such trade-offs by manipulating the level of predator-avoidance behaviour and immune function in the freshwater snail Lymnaea stagnalis (Gastropoda: Pulmonata). Our results show that predator avoidance and immune function had clear costs in terms of reproduction and survival. Further, we show that increased levels of predator-avoidance behaviour reduced the snails' ability to defend against potential pathogens. Predator-avoidance behaviour may thus have the additional indirect cost of reduced immunocompetence and increased susceptibility to pathogens. Our results suggest that ecological factors (e.g. predator density) may considerably modify the expression and costs of immune defences. PMID- 10687824 TI - Density-dependent prophylaxis in the mealworm beetle Tenebrio molitor L. (Coleoptera: Tenebrionidae): cuticular melanization is an indicator of investment in immunity. AB - If there are costs involved with the maintenance of pathogen resistance, then higher investment in this trait is expected when the risk of pathogenesis is high. One situation in which the risk of pathogenesis is elevated is at increased conspecific density. This paper reports the results of a study of density dependent polyphenism in pathogen resistance and immune function in the mealworm beetle Tenebrio molitor. Beetles reared at high larval densities showed lower mortality when exposed to a generalist entomopathogenic fungus and a higher degree of cuticular melanization than those reared solitarily. The degree of cuticular melanization was a strong indicator of resistance, with darker beetles being more resistant than lighter ones regardless of rearing density. No differences were found between rearing densities in the levels of phenoloxidase, an enzyme key to the insect immune response. The results show that pathogen resistance is phenotypically plastic in T. molitor, suggesting that the maintenance of this trait is costly. PMID- 10687825 TI - Intra-host competition between nef-defective escape mutants and wild-type human immunodeficiency virus type 1. AB - Various forms of nef genes with deletions at conserved positions along the sequence have been reported to persist in human immunodeficiency virus type 1 infected patients. We investigate the forces maintaining such variants in the proviral population. The main selection pressures are preservation of function and host immune response. The crippled Nef protein might have fewer epitopes, and as such be less visible to the specific immune response, but it will lose some function. Does a trade-off between avoidance of the immune response and loss of function explain the dynamics of the crippled virus found in the patients? To answer this question, we formulated a deterministic model of the virus-host interactions. We found that when the crippled protein presents few epitopes and suffers little loss of function, the two viral types can coexist. Otherwise, the wild-type comes to prevail. The mutant form might initially dominate, but as the selective pressure by the CD84+ T cells decreases over the course of infection, the advantage for the crippled form of losing epitopes disappears. Hence, we go from a situation of coexistence of wild-type and mutant, to a situation of only full-length nef. The results are discussed in the context of the suggested use of live attenuated vaccines having deletions in nef. PMID- 10687826 TI - Empirical evidence for differential organ reductions during trans-oceanic bird flight. AB - Since the early 1960s it has been held that migrating birds deposit and use only fat as fuel during migratory flight, with the non-fat portion of the body remaining homeostatic. Recent evidence from field studies has shown large changes in organ sizes in fuelling birds, and theory on fuel use suggests protein may be a necessary fuel during flight. However, an absence of information on the body condition of migrants before and after a long flight has hampered understanding of the dynamics of organs during sustained flight. We studied body condition in a medium-sized shorebird, the great knot (Calidris tenuirostris), before and after a flight of 5400 km from Australia to China during northward migration. Not only did these birds show the expected large reduction in fat content after migration, there was also a decrease in lean tissue mass, with significant decreases in seven organs. The reduction in functional components is reflected in a lowering of the basal metabolic rate by 42% [corrected]. Recent flight models have tried to separate the 'flexible' part of the body from the constant portion. Our results suggest that apart from brains and lungs no organs are homeostatic during long-distance flight. Such organ reductions may be a crucial adaptation for long distance flight in birds. PMID- 10687827 TI - An extension to the hypothesis of the asynchrony of visual consciousness. AB - An existing hypothesis states that visual consciousness is made up of 'microconsciousnesses' occurring asynchronously in several sites of the visual system of the brain with no need for direct means of binding. We extend this hypothesis to define what qualifies a neural activity for generating an element of consciousness to distinguish it from one that does not. We argue that, for these separate neural activities to represent elements of a compound sense of consciousness, they each need the support of sites that unconsciously process an important attentional referent and that it is the commonality of such referents in differing sites which bring the microconsciousnesses together. We consider the broader implications of this extended hypothesis for other sensory modalities and mental imagery. PMID- 10687828 TI - Seminal fluid causes temporarily reduced egg hatch in previously mated females. AB - In Drosophila, male accessory gland fluid (seminal fluid) has multiple effects on the female's reproductive efficiency. Here, we show the effect of seminal fluid on rate of egg hatch immediately following mating. Singly mated females were remated to two classes of sterile males, one with seminal fluid and one without seminal fluid. Transfer of seminal fluid results in a strong reduction in egg hatch shortly after the mating. Also, it is shown that remating with normal males causes an immediate reduction of egg hatch followed by recovery to normal egg hatch. In all cases, unhatched eggs contained no sperm. These results are consistent with a role for seminal fluid in sperm competition, mediated by incapacitation or inefficient use of resident sperm. PMID- 10687829 TI - Intrapartum fetal distress and magnesium sulfate. AB - OBJECTIVE: Intrapartum fetal distress is an obstetric emergency traditionally managed by immediate delivery by either the vaginal route or cesarean section. However, there is usually time to attempt intrauterine resuscitation. The purpose of this study was to report the utilization of magnesium sulfate for intrauterine resuscitation. METHOD: Twenty-one fetuses received magnesium sulfate 4-g intravenous bolus in mothers awaiting cesarean section for fetal distress in the labor room of the Complejo Hospitalario Metropolitano de la Caja de Seguro Social de Panama, from March through August 1997. Fetal distress in labor was defined as the presence of repetitive late decelerations, persistent loss of baseline variability, severe variable decelerations, or bradycardia. RESULTS: Twenty-one fetuses received magnesium sulfate 4-g intravenous bolus in mothers awaiting cesarean section for fetal distress. Uterine activity ceased in seven patients, diminished in 13 patients and did not change in one. In all cases, but one, there was recovery of the FHR within 4 min; furthermore there was rose reactive of FHR in nine patients. The 1-min Apgar scores were 7 or above in 18 cases and the 5 min Apgar scores were 7 or above in 20 patients. CONCLUSIONS: In summary, magnesium sulfate may be useful in the management of acute intrapartum fetal distress when there is evidence of increased uterine activity. PMID- 10687830 TI - Antenatal ultrasonography for breech delivery. AB - OBJECTIVE: We studied the role of sonography in the management of 370 breech presentations > or = 34 weeks maturity. METHOD: 185 cases had sonographic confirmation of breech presentation prior to the delivery at the Korle Bu Teaching Hospital. A control group of 185 cases did not have prior scanning. RESULTS: Significantly more elective cesarean sections were done in the study group, while the control group had more emergency sections (P = 0.008), and had more traumatic delivery. Birth asphyxia and perinatal mortality were significantly more common in the control group (P < 0.05). CONCLUSION: Sonography done before delivery improved neonatal outcome in breech presentation > or = 34 weeks maturity. PMID- 10687831 TI - Episiotomy in Nigeria. AB - OBJECTIVE: To review the incidence and complications associated with episiotomy and perineal tears at the University of Benin Teaching Hospital, Benin City, Nigeria. METHOD: A retrospective review of all vaginal births conducted in the hospital between January 1997 and December 1998 was undertaken. Vaginal births (1345) were reviewed. RESULT: The prevalence of episiotomy in the hospital during the period was 46.6%. Over 90% of primigravid parturients had episiotomy. The incidence of episiotomy decreased with increasing parity, while the incidence of spontaneous vaginal tears increased with parity. As compared with perineal tears, episiotomy was associated with a statistically significant increased risk of wound breakdowns requiring secondary resuturing. When controlled for parity, breech births, forceps and vacuum delivery were more likely to lead to episiotomy, compared to spontaneous vertex delivery occurring at term. CONCLUSION: A policy of systematic reduction in the incidence of episiotomy can be pursued in this hospital. Greater attention needs to be paid to selection of women to undergo episiotomy, the prevention of spontaneous perineal tears and the care of episiotomy wounds in this institution. PMID- 10687832 TI - Screening for cervical neoplasia during pregnancy. AB - OBJECTIVE: To evaluate cytology as a screening method for cervical neoplasia in pregnancy and to compare it with cervicography and the acetic acid test (AAT). METHODS: In a large antenatal clinic in South Africa, 842 women were screened utilizing cytology, cervicography and the acetic acid test simultaneously. The proportion of positive results of the different tests were compared and the agreement calculated by the kappa statistic. RESULTS: The mean age of the women was 27 years, and 12.5% smoked. Cytological smears were abnormal (low-grade squamous intra-epithelial lesion and higher degrees of abnormality) in 1.4% of cases, cervicography in 6.3% and the AAT in 14.3% (P = 0.5400). Kappa values were as follows: cytology vs. cervicography 0.01, cytology vs. AAT 0.0 and cervicography vs. AAT 0.2. CONCLUSIONS: As a result of cytology's rather low yield and the small measure of agreement between the tests, cytology should be supplemented by an additional screening test in pregnancy. PMID- 10687833 TI - Treatment outcomes for squamous intraepithelial lesions. AB - OBJECTIVE: To assess the effectiveness of cone biopsy, cryotherapy, laser ablation and the loop electrosurgical procedure in the treatment of squamous intraepithelial lesions. METHOD: Systematic review of randomized controlled trials in subjects who underwent treatment of low- and high-grade squamous intraepithelial lesions with these modalities. Main outcome measures included the following: percent resolution and persistence of a lesion and notable complications for each procedure. RESULT: Pooled rates of resolution for low grade, high-grade, or combined squamous intraepithelial lesions were similar across the different treatment modalities (range 85.2-94.7%), with substantial overlap among the 95% confidence intervals. Significant hemorrhage occurred most frequently in subjects who received cone biopsy (4.6%) (95% CI: 2.15, 6.99), followed by laser ablation (1.75%) (95% CI: 0.70, 2.81), and LEEP (1.35%) (95% CI: 0.24, 2.47). No hemorrhages were reported among subjects who received cryotherapy. Study sample sizes were relatively small. There were no reported cases of progression to invasive cancer, but duration of follow-up (median follow up time for all eligible studies = 12 months) was not sufficient to evaluate long term outcomes. CONCLUSIONS: There were no substantive differences in outcomes regarding persistence and resolution in the treatment of squamous intraepithelial lesions for subjects receiving cone biopsy, cryotherapy, laser ablation, or LEEP. PMID- 10687834 TI - The effects of systemic hormonal replacement therapy on the skin of postmenopausal women. AB - OBJECTIVE: The aim of this study was to determine the effects of hormonal replacement therapy on the skin of postmenopausal women. METHOD: Forty-one postmenopausal women were randomly allocated to receive either hormonal replacement (valerate estradiol--2 mg/day for 21 days and cyproterone acetate--1 mg/day for 10 days) or placebo, both in a cyclic scheme for 6 months. Neither patients nor investigators were aware of the group allocation. Histologic changes were evaluated by skin biopsy of the left upper arm at baseline and after 6 months of treatment, utilizing computerized image analysis to assess the ratio area of epidermis/basement membrane length (AE/BML), ratio area of keratin/basement membrane length (AK/BML) and collagen and elastic fibers content. RESULT: Collagen content of the left upper arm increased after 6 months of treatment only in the hormonal group (+6.49%; P < 0.05). Other parameters did not present any significant alteration after treatment in both groups. CONCLUSION: Hormonal replacement for climacterics increases skin collagen content. PMID- 10687835 TI - Uterine rupture in a multiparous woman during labor induction with oral misoprostol. AB - A multigravida with gestational diabetes, mild pregnancy-induced hypertension and a previous curettage received four doses of misoprostol (100 microg) at three hourly intervals for induction of labor at term. Vaginal delivery of a live healthy baby occurred 1 h after the fourth dose. Hindwaters were bloodstained. Three hours later, she had excessive bleeding. Examination showed that the left lateral uterine wall had ruptured. She recovered after hysterectomy and blood transfusions. PMID- 10687836 TI - Conditions influencing blood pressures in healthy gravidas. AB - The use of 24-h monitoring devices in healthy gravidas confirmed the suspicion that temporary blood pressure elevations relate to ambulation and to emotional upheaval, while the lowest recordings are associated with periods of rest. PMID- 10687837 TI - Treatment for repeat tubal ectopic pregnancy. PMID- 10687838 TI - Emergency postpartum hysterectomy in women with placenta previa and prior cesarean section. AB - The combination of prior cesarean section and placenta previa is an especially ominous risk factor for emergency postpartum hysterectomy and life-threatening bleeding following placental removal. PMID- 10687839 TI - Pregnancy following tubal sterilization: an 11-year survey. PMID- 10687840 TI - Is routine antenatal screening for syphilis useful? AB - The incidence of positive antenatal syphilis serology is low in Benin City. Routine serological screening should continue with the inclusion of confirmatory treponemal antigen tests. PMID- 10687841 TI - Trends in epidemiology of cervical cancer in Thrace, Greece. PMID- 10687842 TI - Voluntary and involuntary sterilization: denials and abuses of rights. AB - Laws that allow competent persons to make free and informed decisions for sterilization serve their entitlements to reproductive choice. Laws that allow others to consent to sterilization of disadvantaged persons who cannot freely consent risk oppression and denial of human rights. Laws that prohibit competent persons' choices for their own sterilization are comparably oppressive and violative of human rights to decide whether and how often to have children. Whether laws approach sterilization as a procedure done for patients, or to patients, is often ambivalent. Details of laws may indicate their liberating and oppressive potential. Programs offering inducements to persons to be sterilized may assist those who are disadvantaged to achieve their goals, but may appear to coerce those who, through poverty or dependency, cannot resist the inducement. PMID- 10687843 TI - ACOG committee opinion. Quality of laboratory and imaging services: physician responsibility in the age of managed care. Number 222, October 1999. Committee on Practice Management. Committee on Professional Liability. American College of Obstetricians and Gynecologists. PMID- 10687844 TI - ACOG committee opinion. First-trimester screening for fetal anomalies with nuchal translucency. Number 223, October 1999. Committee on Genetics. American College of Obstetricians and Gynecologists. PMID- 10687845 TI - ACOG committee opinion. Tamoxifen and the prevention of breast cancer in high risk women. Number 224, October 1999. Committee on Gynecologic Practice. American College of Obstetricians and Gynecologists. PMID- 10687846 TI - Oocyte-expressed TGF-beta superfamily members in female fertility. AB - Folliculogenesis is regulated by the interplay of extraovarian and intraovarian factors, and the importance of each type of regulation varies depending on the developmental stage of the follicle. Preantral follicle development is regulated predominantly by factors produced locally within the ovary and within the follicle itself. The oocyte has been shown to produce soluble factor(s), which regulate a number of processes in follicular development, including cumulus expansion in the periovulatory period. Members of the TGFbeta superfamily are potent regulators of cell proliferation and differentiation in a number of organ systems, and three members, growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15) and BMP-6 are expressed by the oocyte and may mediate effects attributed to the oocyte. Based on knockout mouse models BMP-6 does not play an essential role in ovarian function, but GDF-9 is absolutely required for preantral follicle development. GDF-9 also alters the periovulatory expression of granulosa cell genes and stimulates cumulus expansion. Although BMP 15 is expressed identically to GDF-9, its role in regulating ovarian function is still unknown. This review examines the similarities and differences in sequence, expression, and function of the oocyte-expressed TGFbeta family members with respect to regulating folliculogenesis. PMID- 10687847 TI - Osteocyte function, osteocyte death and bone fracture resistance. AB - The function of the most numerous cell in bone, the osteocyte, has until recently been mysterious and at times controversial. There is now an emerging consensus that osteocytes modulate signals arising from mechanical loading and so direct the appearance and disappearance of bone tissue at the microscopic level, which allows bone as an organ both to grow and to adapt efficiently to the body's mechanical needs for strength with lightness. Osteocytes appear to use some molecular signalling pathways that are familiar from other tissues, such as the generation of nitric oxide and prostaglandins as well as directing cell-cell communication via gap junctions. They may also direct the removal of damaged or redundant bone through mechanisms linked to their own apoptosis or via the secretion of specialised cellular attachment proteins such as osteopontin. Osteocytes possess receptors for parathyroid hormone/parathyroid hormone related peptide and both oestrogen receptors alpha and beta. They also express molecules which in nerve cells are involved with glutamate neuro-transmission. At least some of these receptors and their ligands may regulate osteocyte apoptosis and modulate osteocyte signalling. PMID- 10687848 TI - Cloning and characterization of a novel retinoid-inducible gene 1(RIG1) deriving from human gastric cancer cells. AB - Retinoids exert wide-spectrum anti-tumor activities, which are mediated via the induction of growth arrest, differentiation or apoptosis. To determine whether the effects of retinoids are mediated by specific gene activation or repression, SC-M1 CL23 gastric cancer cells, pretreated with either vehicle alone or all trans retinoic acid (10 microM) for 1 day, were analyzed using the technique of differential display. A novel retinoid-inducible gene 1 (RIG1) was isolated. The full-length RIG1 cDNA contained 768 base pairs and encoded a protein of 164 amino acids with a molecular weight of 18 kDa. The RIG1 gene was ubiquitously expressed in normal tissue, and its expression was positively associated with cellular density. Nucleotide sequence analysis demonstrated that the RIG1 gene was similar to a recently-isolated TIG3 gene, and displayed 54% nucleotide sequence homology with a type II tumor suppressor gene H-REV-107-1. RIG1 cDNA, however, contained an extra 32 base pairs located at its 5' end and revealed three base pair differences for the remaining sequences leading to two amino acids substitution between the two encoded proteins. All-trans retinoic acid increased the level of RIG1 mRNA in a time- and concentration-dependent manner in SC-M1 CL23 gastric cancer cells. This was not observed for the H-REV-107-1 gene. The RIG1 regulation was related to cellular retinoid sensitivity. Both retinoic acid receptor alpha- and retinoic acid receptor gamma-selective agonists increased RIG1 mRNA level, and the retinoid x receptor-selective agonist potentiated this regulation. In conclusion, the cDNA of a novel retinoid-inducible gene RIG1 has been cloned. This gene is regulated by retinoic acid through the heterodimer of retinoic acid receptor and retinoid x receptor. PMID- 10687849 TI - Mechanisms underlying the steroidogenic synergy of insulin and luteinizing hormone in porcine granulosa cells: joint amplification of pivotal sterol regulatory genes encoding the low-density lipoprotein (LDL) receptor, steroidogenic acute regulatory (stAR) protein and cytochrome P450 side-chain cleavage (P450scc) enzyme. AB - Growth of ovarian Graafian follicles and cytodifferentiation of granulosa and theca cells are regulated by gonadotropins, sex steroids and peptidyl growth factors. For example insulin and intraovarian insulin-like growth factor type I (IGF-I) may amplify the actions of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) in promoting biochemical luteinization and enhancing steroidogenesis. To explore further the notion of interactions between insulinomimetic peptides and LH and to examine the associated mechanisms, we have established porcine granulosa cells in monolayer culture for 48 h in 3% serum with insulin (1 microg/ml), estradiol (0.5 microg/ml), and follicle stimulating hormone (FSH, 5 ng/ml) to allow cell anchorage, facilitate in vitro cytodifferentiation and confer LH responsiveness. To limit any carry-over effects of serum, granulosa cells were stabilized overnight in serum-free medium. Studies were then initiated to assess the impact of insulin on the dose-responsive actions of LH. A maximally effective concentration of insulin (1 microg/ml) synergistically augmented LH's dose-dependent ampilification of progesterone and cAMP accumulation; viz. by approximately twofold (progesterone) and approximately 2.5-fold (cAMP) above that observed in maximally LH-stimulated cultures (P < 0.001). Mechanistically, insulin significantly enhanced the sensitivity of granulosa cells to LH's drive of cAMP accumulation [ED50 for LH 61 +/- 14 ng/ml (control) vs. 10 +/- 1.0 ng/ml (insulin) (P < 0.01)]. Insulin also augmented the maximal stimulatory effect of LH; i.e. LH efficacy rose from 6.5 +/- 0.4 to 17 +/ 1.4 (pmole cAMP/microg DNA/48 h; P < 0.001). Insulin dose-response analysis showed that insulin alone minimally elevated basal, but significantly heightened LH's stimulation of progesterone and cAMP accumulation at (insulin) concentrations as low as 3-10 ng/ml. The molecular mechanisms underlying insulin and LH's synergy were assessed by RNase protection assays with (porcine) cRNA probes encoding the low density lipoprotein receptor (LDL-R), Steroidogenic Acute Regulatory Protein (StAR), P450 cholesterol sidechain cleavage enzyme (P450scc) and (as a possible negative control) Sterol Carrier Protein 2 (SCP-2) [data normalized to constitutive 18S rRNA]. Non linear least-squares analysis was applied to confirm or refute an hypothesis of interactive synergy between LH and insulin on gene expression. LH and insulin alone exerted no effect on StAR message accumulation, and LH alone minimally stimulated P450scc and LDL-R mRNA's accumulation at 48 h. In contrast, insulin in combination with LH augmented StAR mRNA concentrations by approximately 5-10-fold and stimulated LDL-R message levels by threefold above the respective maximally LH-driven values (P < 0.01). Maximal P450scc mRNA expression was enhanced twofold by cotreatment with LH and insulin compared with maximal LH-treated cultures. In contrast SCP-2 mRNA accumulation remained unaffected by any treatment. In summary, we have used a serum-free, in vitro differentiated porcine granulosa cell culture system to assess regulatory interactions between the disparate first messengers, LH and insulin. We observe marked LH-insulin steroidogenic synergy after 48 h of joint hormonal stimulation, and further clarify that the mechanism(s) of synergy include augmentation of cAMP production and increased steady-state concentrations of transcripts of key sterol-regulatory genes; namely, LDL-R, StAR, and P450scc, but not SCP-2. Since the encoded products of these genes variously control sterol substrate uptake, delivery to and utilization in mitochondrial steroidogenesis, we speculate that the concerted actions of insulin-like peptides and LH may contribute to steroidogenic differentiation during the later stages of follicular maturation and the granulosa-luteal cell transition. PMID- 10687850 TI - Expression of GnRH receptor gene in human ectopic endometrial cells and inhibition of their proliferation by leuprolide acetate. AB - The present study was conducted to investigate whether GnRH-receptor (GnRH-R) gene is expressed in endometriosis ovarian implants and whether a GnRH-analogue (GnRH-a) may exert an effect on endometriosis cell proliferation in vitro. The presence of GnRH-R transcripts in ovarian endometriosis cells was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and further confirmed by Southern blot analysis. GnRH-R mRNA was detected in all the 13 samples examined. In contrast, GnRH-R transcripts were not detectable in endometriosis-free peritoneal tissue. In the second part of the study, endometriosis cells were cultured for 9 days with different doses of leuprolide acetate (ranging from 0 to 10(-5) M). In 4 out of 13 cases, a significant anti-proliferative effect was observed at doses of leuprolide acetate ranging from 10(-9) to 10(-5) M. In one case, a significant inhibition of cell proliferation was observed only at 10(-5) M leuprolide acetate concentration. In contrast, the GnRH-a did not affect cell growth, regardless of the expression of GnRH-R transcripts and the given doses, in the remaining 8 experiments. To date, this is the first evidence indicating that GnRH-R mRNA is expressed in human ovarian endometriomas. Moreover, the inhibition of endometriosis cell proliferation induced by the GnRH-a in vitro suggests that, at least in some cases, this compound might exert a direct effect on endometriosis lesions. PMID- 10687852 TI - EGF activates highly selective estrogen-responsive reporter plasmids by an ER independent pathway. AB - Epidermal growth factor (EGF) mimics the effects of estrogen on some cells, suggesting that it may activate the estrogen receptor (ER). We examined the ability of EGF to increase expression of several different estrogen-responsive luciferase reporters in MCF-7 breast cancer cells. Although EGF increased reporter activity, this effect was not inhibited by estrogen antagonists and was not dependent on estrogen response elements in the reporter plasmid. Similar results were obtained in BG-1 (ovarian) and Ishikawa (uterine) cells. In ER negative JEG-3 cells, EGF, but not estradiol, increased reporter activity in the absence of transfected ER. The estrogen antagonist ICI 182780 blocked the ability of estradiol, but not EGF, to stimulate proliferation of T47D breast cancer cells, suggesting that the mitogenic effects of EGF are not mediated by ER. EGF does not appear to activate ER-mediated transcription in these experimental systems, although crosstalk between the estrogen and EGF signaling pathways may occur by other mechanisms. PMID- 10687851 TI - Effect of cyclic adenosine 3',5'-monophosphate and protein kinase A on ligand dependent transactivation via the vitamin D receptor. AB - We examined the effects of cyclic adenosine 3',5'-monophosphate (cAMP) and protein kinase A (PKA) on the ligand-dependent transactivation mediated via the 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) receptor (VDR). A human VDR expression plasmid was transfected into HeLa, Saos-2 and MG63 cells with a luciferase reporter gene construct containing the vitamin D responsive element. With the addition of 0.5 mM 8 bromo-cAMP, the response to 1,25(OH)2D3 was suppressed to 61 and 78% in the HeLa and Saos-2 cells, respectively. The suppressive effect of 8 bromo-cAMP was observed without the introduction of the VDR expression plasmid in the MG63 cells. In the HeLa cells the co-expression of PKA reduced the ligand inducible transactivation to 61% and the fold induction by 1,25(OH)2D3 to 89% of that without PKA. The CREB binding protein (CBP) was recently reported to integrate the intracellular signals via the cAMP/PKA cascade and nuclear hormone receptors. However, the suppressive effect of cAMP was not influenced by the co expression of CBP. Lastly, we introduced point mutations at possible PKA phosphorylation sites into the VDR expression vector at serine-172 and threonine 175, but both mutant receptors still exhibited reduced transactivation with the co-expression of PKA. These results indicate that the phosphorylation of proteins other than the VDR may also be involved in the inhibitory effect mediated by the cAMP/PKA cascade. PMID- 10687853 TI - Links between Fer tyrosine kinase expression levels and prostate cell proliferation. AB - In our cloning strategy to identify tyrosine kinases implicated in the regulation of prostate growth, the dog fer cDNA was obtained and shown to be highly homologous to known fer cDNAs. Using a polyclonal Fer antibody directed against a C-terminal peptide, we studied its associations with cortactin, beta-catenin and p120Cas in human prostate carcinoma PC-3 cells. In contrast to previous reports, no interactions were observed. To assess its functional role, fer cDNA constructs were transfected in PC-3 cells. Antisense clones exhibiting a marked diminution of Fer expression had a reduced growth rate (doubling time of 29 vs. 42 h) and were unable to form colonies in soft agar. In agreement with these results, Fer protein expression was linked to human prostatic proliferative diseases, with enhanced levels in extracts from cancer tissues as compared to those from normal and hyperplastic ones, and was also expressed in the human prostate carcinoma cell lines DU145 and LNCaP. In the dog model, Fer expression was up-regulated in dividing versus resting prostate epithelial cells in vitro, and also in vivo when basal cell hyperplasia and metaplasia was induced by estrogen after castration. Minimal effects were observed when renewing the luminal epithelium with androgens. Taken together, these results show that Fer expression is associated with prostate cell proliferation and enhanced in prostate cancer. PMID- 10687854 TI - Tumour necrosis factor-alpha exerts dual effects on human adipose leptin synthesis and release. AB - The acute and chronic effects of tumour necrosis factor-alpha (TNF-alpha) on leptin production by human preadipocytes, differentiated preadipocytes, and mature adipocytes have been examined by competitive RT-PCR of leptin mRNA and by western blotting. In preadipocytes, secreted leptin was detectable after 5-day incubation in differentiation medium and this increased 4-fold by day 20. TNF alpha blocked leptin synthesis during differentiation. In differentiated preadipocytes and mature adipocytes, TNF-alpha treatment resulted in time dependent decreases in mRNA for leptin and glycerol-3-phosphate dehydrogenase (G3PD). In contrast, TNF-alpha (4-8-h treatment) resulted in a 4-fold increase in leptin release. This effect was lost at 24 h and leptin accumulation in culture medium was decreased 24-48 h after TNF-alpha addition. We conclude that TNF-alpha stimulates the release of preformed leptin from human mature adipocytes and existing differentiated preadipocytes, which may contribute to obesity/infection linked hyperleptinemia, and that TNF-alpha inhibits leptin synthesis via inhibition of preadipocyte differentiation and induction of adipocyte dedifferentiation. PMID- 10687855 TI - Mouse growth hormone transcription factor Zn-16: unique bipartite structure containing tandemly repeated zinc finger domains not reported in rat Zn-15. AB - Rat Zn-15 is a transcription factor activating GH gene expression by synergistic interactions with Pit-1, named for 15 DNA-binding zinc fingers, including fingers IX, X, and XI that are responsible for GH promoter binding. In this study, a mouse cDNA for Zn-15 was characterized. The predicted 2192-amino acid mouse protein is 89% identical to rat (r) Zn-15 overall, and is 97% similar in the C terminal domain necessary for binding the GH promoter. However, the mouse cDNA encodes 16 zinc fingers, and sequences of rZn-15 pituitary cDNAs were the same as the mouse (m) Zn-16; the rat sequence in GenBank has a one nucleotide offset of a 17-bp segment in the finger V region. The mouse and corrected rat sequences contain four tandemly repeated fingers in the N-terminus, each separated by seven amino acids, typical of zinc finger proteins of the transcription factor IIIA type. Analysis of mZn-16 expression by RT-PCR showed that the mRNA is, produced at similar levels in normal and GH-deficient Ames dwarf (Prop-1 ) mouse pituitaries at postnatal day 1. Mouse Zn-16 mRNA also was detected by ribonuclease protection assay in the pre-somatotrophic mouse cell line GHFT1-5. The Zn-16 protein is bipartite in that the N-terminal half displays tandem spacing typical of most zinc finger proteins, while the C-terminal portion contains long linkers between fingers that cooperatively bind to a DNA response element. Expression in early postnatal pituitary and in pre-somatotrophic cells suggests that Zn-16 could play a role in pituitary development prior to somatotroph differentiation. PMID- 10687856 TI - Expression and regulation of melanocortin receptor-5 (MC5-R) in the bovine adrenal cortex. AB - Among the five members of the melanocortin receptor (MC-R) family, MC2 and MC5 are expressed in peripheral tissues. The receptor MC2 (ACTH receptor) almost exclusively expressed in the adrenal cortex whereas MC5-R is expressed in several organs including the adrenal cortex. Both receptors bind ACTH and activate adenylate cyclase. The aim of this work was to study the spatial distribution of MC5-R among the different zones of the bovine adrenal cortex and to analyze the regulation of its expression by its own ligands, ACTH and alpha-MSH and by angiotensin II (AII). Using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and RNase protection assay, MC5-R was detected only in the glomerulosa zone whereas MC2-R was present in both glomerulosa and fasciculata zones of adult adrenal cortex. Treatments by ACTH, alpha-MSH, or AII increased the MC5-R mRNA level in glomerulosa cells by factors 7, 5, and 4.5, respectively. However, although potentially regulated by hormones, MC5-R is expressed at a level at least 100 times less than MC2-R, suggesting that MC5-R expression might only be at trace levels in grown adults, but could be much higher during embryogenesis. PMID- 10687857 TI - Resistance to thyroid hormone (RTH) syndrome reveals novel determinants regulating interaction of T3 receptor with corepressor. AB - Thyroid hormone receptors (T3Rs) both repress and activate gene transcription by interacting with auxiliary factors denoted corepressors and coactivators. Resistance to thyroid hormone (RTH) syndrome in humans is manifested as a failure to respond properly to elevated circulating thyroid hormone. RTH syndrome has been mapped to T3Rbeta mutations that alter the transcriptional properties of the receptor, resulting in a dominant negative phenotype. We report here a characterization of a series of RTH mutant T3Rs that exhibit unusual interactions with corepressor. Two mutations in receptor helix 11 (delta430, delta432) greatly enhance the ability of the mutant receptors to bind to corepressor. A distinct mutation, V264D, in an 'omega loop' region of the receptor, impairs corepressor release but does not fully eliminate the ability to recruit coactivator. These mutations reveal novel determinants that regulate the interaction of the T3R with important ancillary cofactors, and that are disrupted in a human endocrine disease. PMID- 10687858 TI - An alternatively spliced rat mineralocorticoid receptor mRNA causing truncation of the steroid binding domain. AB - We attempted to clone the putative 11-dehydrocorticosterone receptor by RT-PCR with two degenerate primers from highly homologous regions of the DNA and steroid binding domains of the receptor subfamily. In doing so, we have identified an alternatively spliced variant mRNA of the rat mineralocorticoid (MR) with a ten bp deletion in the C-terminal steroid binding domain. This deletion results in a truncated MR receptor of 807 amino acids in comparison to the wild type of 981 amino acids. The deletion variant was expressed in colon, kidney, heart, liver, aorta and brain tissues. The relative abundance of the deletion variant compared to the wild type MR was estimated to be 6% in rat kidney and 4% in hippocampus. This deletion was also detected in human kidney by RT-PCR. Site-directed mutagenesis was used to create the eukaryotic expression plasmid pCR3-rMRdel10 from the wild type for a transactivation assay using the luciferase reporter system in CV-1 cells. The deletion variant had the same baseline transactivation activity as the wild type MR, but did not respond to aldosterone or corticosterone stimulation. Co-transfection of MR with the deletion variant had no significant effect on transactivation activity of the MR, indicating that the deletion variant is unlikely to serve as a negative regulator of MR function. PMID- 10687859 TI - Effect of PRL on MAPK activation: negative regulatory role of the C-terminal part of the PRL receptor. AB - Prolactin induces cell proliferation and cell differentiation through well-known MAPK Erk, and JAK2/STAT5 pathways depending on the cell line. The aim of the present study was to delineate the functional domains of the PRL receptor involved in PRL induced MAPK regulation. Using various PRL-R mutants of the cytoplasmic domain we found, that the membrane proximal domain is necessary for PRL induced MAPK activation and that the C-terminal part of the receptor exerts a negative regulatory role. A pharmacological approach, using different types of inhibitors, provided evidence that PRL induced MAPK activation requires both a MEK dependent pathway and a PI3K dependent pathway. The negative regulation induced by the carboxy-terminal part of the receptor involves a combination of tyrosine phosphatases and serine/threonine phosphatases as concluded from the actions of the phosphatase inhibitors: pervanadate, PAO and okadaic acid. The mechanism by which these phosphatases are recruited or are induced by the last 141 cytoplasmic residues of the receptor remains to be determined. Finally the negative regulatory role of the carboxy-terminal part of the receptor, first demonstrated in the present study, is discussed in terms of the regulation of different effects of PRL on growth and differentiation. PMID- 10687860 TI - Relaxin-like factor (RLF) mRNA expression in the fallow deer. AB - Employing RT- and RACE-PCR on RNA isolated from testicular tissue, we have cloned the coding cDNA sequence for the RLF, also known as Insl3, of the fallow deer. The RLF coding sequence consisted of 396 bp encoding a peptide of 131 amino acids and shared highest homology with bovine, sheep and goat RLF. Northern analysis revealed a single 0.9 kb transcript in the deer testis. There is only one RLF gene in the deer genome. Nonradioactive in situ hybridization revealed the Leydig cells to be the sole source for RLF mRNA in the deer testis. In the non-pregnant uterus, RLF transcripts were located in the luminal and glandular epithelium of the endometrium. Within the ovary of the pregnant doe, follicular theca interna cells and the corpus luteum expressed RLF transcripts. In uteroplacental tissues, luminal and glandular epithelium, fetal uninucleate and binucleate trophoblast cells (BNC) of the basic villous trophoblast layer expressed RLF mRNA. BNC located at the apical trophoblast layer or the tip of the fetal villus were devoid of RLF transcripts. Pseudostratified trophoblast cells at the base of fetal villi coexpressed RLF mRNA and immunoreactive MHC class Ib molecules. PMID- 10687861 TI - Expression and regulatory function of the transcription factor Sp1 in the uterine endometrium at early pregnancy: implications for epithelial phenotype. AB - The uterus during early pregnancy synthesizes a complex array of signaling molecules with specific spatial and temporal modes of expression and which are critical for embryo implantation and subsequent development. The mechanism(s) underlying the differential pattern of synthesis of these pregnancy-associated proteins is not understood very well. The present study evaluated the expression and trans-activation potential of the transcription factor Sp1 in the early pregnancy porcine endometrium to determine its temporal and functional association with the endometrial epithelial-specific genes encoding the transplacental iron-transport protein uteroferrin (UF) and an Sp-family member, basic transcription element-binding (BTEB) protein. Two identical Sp1 clones (717 bp) were isolated from a porcine endometrial cDNA library by polymerase chain reaction (PCR). The nucleotide sequence of these clones encodes a partial protein sequence of 238 amino acids encompassing the Zn-finger region and had significant identities with the corresponding regions in the rat and human proteins. By using a specific antibody raised against human Sp1, porcine endometrial Sp1 was found to exhibit a molecular weight of 110 kDa, was localized predominantly in the nuclei of glandular and luminal epithelial cells, and appeared to exist as a phosphorylated protein. Northern blot analysis demonstrated three distinct size transcripts of approximately 3.5, 5, and 8 kb for endometrial Sp1. The expression of Sp1 mRNA and protein, determined by RT-PCR and by its ability to bind Sp1 consensus motif in gel mobility shift assays, respectively, overlapped with, but did not parallel that of UF mRNA during early pregnancy. The effect of increased Sp1 expression on UF gene promoter activity was examined using a human Sp1 expression vector that was transiently transfected into primary cultures of pig endometrial glandular epithelial cells. Sp1 increased (P < 0.05) the promoter activities of various UF promoter-Luciferase reporter constructs by 2 to 4-fold, over those transfected with empty expression vector. Co-transfection of a BTEB expression vector with the Sp1 expression vector modified the effect of Sp1 on UF promoter activity in the shortest construct. These results suggest that Sp1 mediates the regulation of endometrial epithelial gene expression during pregnancy, and that this function is likely altered in vivo by co-expression of other family members, including BTEB. PMID- 10687862 TI - Partial characterization of the CCAAT box in the promoter of the hLGFBP-1 gene: interaction with negatively acting transcription factors in decidualized human endometrial stromal cells. AB - The CCAAT cis-element and its adjacent DNA sequence (-82 to -52 bp) in the human insulin-like growth factor binding protein-1 gene (IGFBP-1) promoter are active in both decidualized human endometrial stromal cells and HepG2 cells. In HepG2 cells, CCAAT activity is mediated by interacting with hepatocyte nuclear factor, HNF-1. In endometrial cells, this region is protected by the nuclear extracts of endometrial decidual cells, however, the transactivator which interacts with the region has not been identified. This study was carried out to characterize and identify the stromal/decidual nuclear proteins that interact with the IGFBP-1 CCAAT motif. Gel shift analysis showed that the CCAAT motif (-82 to -52 bp) formed three specific complexes (CI, CII, and CIII) by extracts from human endometrial decidual or stromal cells. The intensity of CIII formed by the nuclear extracts of decidual cells was less compared to that formed by stromal cells whereas CI/CII was found to be opposite. To evaluate the transcription factors that bind to this region, a number of known CCAAT binding proteins were tested. Among them, the CCAAT binding proteins NF-Y (alpha2(1) collagen promoter CCAAT binding protein) and CBF (hsp70 promoter CCAAT binding protein), were characterized by the gel shift assay. The NF-Y consensus binding sequence (the alpha2(1) collagen promoter) and NF-YA,B antibody abolished or shifted CIII. Although the CBF consensus binding sequence (the hsp70 promoter) eliminated all three complexes, the antibody to CBF had no effect on all three complexes. The nuclear extracts of the endometrial stromal/decidual cells did not form a band corresponding to the HNF-1/CCAAT complex. These results indicate that the CCAAT motif binds to NF-Y and the CI/CII binding protein (remains to be identified) but not HNF-1 in endometrium. Systematic mutation in the CCAAT motif showed that NF Y(CIII binding protein) bound to the 12 bp sequence GGCGCTGCCAAT(-79 to -68 bp) and the CI/CII binding protein bound to 9 bp, TGCCAATCA(-74 to -66 bp). These findings indicate that the CCAAT motif is a composite element. The CCAAT mediated function was analyzed in decidualized endometrial stromal cells. Mutations in the CCAAT motif increased the promoter activity. The maximum activity was found in mutants which abolished the NF-Y complex. The CCAAT core sequence mutants in which both CIII and CI/CII were abolished, also increased the promoter activity. Results indicated that NF-Y and the CI/CII binding protein, yet to be identified, interact with the composite CCAAT element in the IGFBP-1 promoter to repress the promoter activity in endometrial decidual cells. PMID- 10687863 TI - Permissive effect of thyroid hormones on induction of rat colonic Na+ transport by aldosterone is not localised at the level of Na+ channel transcription. AB - The interrelationship between thyroid hormones and aldosterone has been examined in the regulation of rat colonic amiloride-sensitive Na+ transport which translocates Na+ through apical amiloride-sensitive Na+ channels and basolateral Na+, K+-ATPase. Electrogenic Na+ transport was measured in an Ussing chamber by the short-circuit current and identified by Na+ channel blocker amiloride. Na+ pumping activity of the basolateral Na+,K+-ATPase was investigated in nystatin treated epithelium by measuring the equivalent short-circuit current after addition of mucosal Na+. The abundance of mRNA coding for alpha, beta and gamma subunits of the Na+ channel (rENaC) was estimated using Northern blot analysis. Hyperaldosteronism was induced by a low-salt diet and hypothyroidism by methimazole. The low-Na+ diet induced electrogenic Na+ transport in euthyroid rats but its effect was almost completely inhibited in hypothyroid animals even if the plasma concentration of aldosterone was high enough to stimulate this transport pathway both in euthyroid and hypothyroid rats. A kinetic study of the basolateral Na+,K+-ATPase revealed a decrease of Na+ transport capacity in hypothyroid rats kept on the low-Na+ diet in comparison with euthyroid animals fed the same diet. No significant differences in steady-state levels of alpha, beta and gamma rENaC mRNA were detected between euthyroid and hypothyroid rats. These data suggest that hypothyroidism decreases the efficacy of the basolateral Na+ pump but fails to inhibit it completely even though it inhibits the transepithelial electrogenic Na+ transport in response to aldosterone. We conclude that the permissive effect of thyroid hormones on the induction of electrogenic Na+ transport by aldosterone is localised beyond the transcriptional step of Na+ channel regulation. PMID- 10687864 TI - Temporal expression and T3 induction of thyroid hormone receptors alpha1 and beta1 during early embryonic and larval development in zebrafish, Danio rerio. AB - The effects of thyroid hormones on metabolism and development are mediated by thyroid hormone receptors (TRs). We report the cloning and characterization of a TR beta1 cDNA from zebrafish. Southern blot analysis revealed that there is a single genomic locus for the TR beta gene, while the TR alpha gene potentially has two loci. Multiple TR alpha and TR beta transcripts were detected in adult tissues. Using a semiquantitative RT-PCR assay, zygotic expression of TR alpha1 and TR beta1 were shown to occur before the midblastula transition stage. In transiently transfected HeLa cells, TR alpha1 displayed constitutive transactivation in the absence of ligands, which was slightly enhanced by triiodothyronine (T3). The transactivating activity of TR beta1 was strictly ligand-dependent and repressed in the absence of T3. Finally, the T3 induction of TR alpha1 and TR beta1 mRNAs was demonstrated in zebrafish embryos and larvae. The auto-induction of TR alpha1 and TR beta1 may serve a regulatory role during the embryonic and larval development of zebrafish. PMID- 10687865 TI - Perinatal toxicology of Ruta chalepensis (Rutaceae) in mice. AB - Dried leaf infusions of Ruta chalepensis L. (Rutaceae), 'rue', 'ruda', were found to cause perinatal changes in mice, at daily doses of 0.16, 0.80 and 1.60 g/kg, administered p.o. from 1 to 14 days post coitum. Significant decreases in the appearance time of physical signs, righting reflex and cliff avoidance together with minus scores in string test and swimming ability were observed. Moreover, histological studies showed progressive angiogenic development on placenta blood supply and weakness at blood barrier in brain, thymus and pery-lymph vestibule. We found out that the results tend to confirm the embryotoxic effect of the plant and its harmful use. PMID- 10687866 TI - Effect of Tephrosia vogelii and Justicia extensa on Tilapia nilotica in vivo. AB - According to our and other ethnobotanic studies (Walker, R., 1951. Une Nouvelle Legumineuse du Gabon servant a narcotiser le poisson. Rev. Bot. Appl. 31, 327; Walker, R., Sillans, R., 1961. Les plantes utiles du Gabon. Encyclopedie Biologique. P. Chevalier, Paris; Halle, N., 1970. Flore du Gabon 17, Famille des Rubiacees. Museum National d'Histoire Naturelle, Paris; Mounzeo, H., et al., 1997. Quelques plantes utilisees comme poisons de peches chez les Punu du Gabon. Le Flamboyant 44. Decembre, Bulletin de Liaison des membres du reseau 'Arbres Tropicaux'), Tephrosia vogelii and Justicia extensa are two plants whose leaves are particularly used for the catching of fish in Gabonese rivers. The leaf extracts of those plants have been tested on Tilapia nilotica in order to observe their toxicity. At a given dose, the small fish are the first to be poisoned. This toxicity is more important for J. extensa and increased in a dose-dependent manner. After boiling for 90 min, those leaf extracts and rotenone (10(-6) M) taken as a control retain their toxicity at high dose (625 mg/l), although the latency period is higher. With the same temperature condition, at weak doses (37.5 and 62.5 mg/l), T. vogelii loses its toxicity, whereas J. extensa preserves it at 62.5 mg/l. As shown in our results, the fact that the extracts preserve their toxicity at high dose after boiling requires particular attention be given to the doses used for fishing and to the type of plants used. PMID- 10687867 TI - CNS acetylcholine receptor activity in European medicinal plants traditionally used to improve failing memory. AB - Certain Lamiaceous and Asteraceous plants have long histories of use as restoratives of lost or declining cognitive functions in western European systems of traditional medicine. Investigations were carried out to evaluate human CNS cholinergic receptor binding activity in extracts of those European medicinal plants reputed to enhance or restore mental functions including memory. Ethanolic extracts were prepared from accessions of these plants and a number of other species related by genus. Amongst the plant extracts screened for contents able to displace [3H]-(N)-nicotine and [3H]-(N)-scopolamine from nicotinic receptors and muscarinic receptors, respectively in homogenates of human cerebral cortical cell membranes, the most potent extracts, prepared from one accession of Melissa officinalis, three Salvia species and Artemisia absinthium had IC50 concentrations of < 1 mg/ml. The displacement curves of some extracts were comparable with that of carbamylcholine chloride, a potent acetylcholine analogue. Choline, a weak nicotinic ligand (IC50 = 3 x 10(-4) M) was found in extracts of all plants studied at concentrations of 10(-6)-10(-5) M. These concentrations could not account for not more than 5% of the displacement activity observed. Some extracts displayed differential displacement at nicotinic and muscarinic acetylcholine receptors, with M. officinalis 0033 having the highest [3H]-(N)-nicotine displacement value and Salvia elegans with the highest [3H]-(N)-scopolamine displacement value. There was also considerable variation in cholinoreceptor interactions between different accessions of a single plant species. Although most plant extracts screened showed some nicotinic and muscarinic activity, only some showed dose-dependent receptor activity typical of materials with genuine cholinergic activity. PMID- 10687868 TI - Enhanced DNA repair, immune function and reduced toxicity of C-MED-100, a novel aqueous extract from Uncaria tomentosa. AB - Female W/Fu rats were gavaged daily with a water-soluble extract (C-MED-100) of Uncaria tomentosa supplied commercially by CampaMed at the doses of 0, 5, 10, 20, 40 and 80 mg/kg for 8 consecutive weeks. Phytohemagglutinin (PHA) stimulated lymphocyte proliferation was significantly increased in splenocytes of rats treated at the doses of 40 and 80 mg/kg. White blood cells (WBC) from the C-MED 100 treatment groups of 40 and 80 mg/kg for 8 weeks or 160 mg/kg for 4 weeks were significantly elevated compared with controls (P < 0.05). In a human volunteer study, C-MED-100 was given daily at 5 mg/kg for 6 consecutive weeks to four healthy adult males. No toxicity was observed and again, WBC were significantly elevated (P < 0.05) after supplement. Repair of DNA single strand breaks (SSB) and double strand breaks (DSB) 3 h after 12 Gy whole body irradiation of rats were also significantly improved in C-MED-100 treated animals (P < 0.05). The LD50 and MTD of a single oral dose of C-MED-100 in the rat were observed to be greater than 8 g/kg. Although the rats were treated daily with U. tomentosa extracts at the doses of 10-80 mg/kg for 8 weeks or 160 mg/kg for 4 weeks, no acute or chronic toxicity signs were observed symptomatically. In addition, no body weight, food consumption, organ weight and kidney, liver, spleen, and heart pathological changes were found to be associated with C-MED-100 treatment. PMID- 10687869 TI - A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part I. Evaluation of the antimalarial activity of plants used by the Chacobo Indians. AB - Thirty extracts of plants traditionally used by the Chacobos, a native community living in the Amazonian part of Bolivia, were screened in vitro and/or in vivo for antimalarial activity. Two of the four species designated as antimalarial, Geissospermum laeve and Maquira coriacea, displayed rather good activity, corroborating their traditional uses. However, they did show a rather high toxicity in vivo. Among twelve species used to cure symptoms relevant to malaria, five showed good activity: Apuleia leiocarpa, Bauhinia guianensis, Nectandra cuspidata, Sparattanthelium amazonum, Tanaecium jaroba. Two species, Qualea paraensis and Sclerolobium aff. guianense, used to treat scabies, showed interesting antimalarial activity in vivo; three other species (Iryanthera laevis, Prunus amplifolia, Pterocarpus aff. amazonum) used for various medicinal purposes, apparently not related with a Plasmodium infection, also showed antimalarial activity. Finally, one species (Derris amazonica) used as a piscicide displayed good in vitro activity, in the same way as one Annonaceae, Guatteria aff. schomburgkiana, used for construction purposes. PMID- 10687870 TI - The search for natural bioactive compounds through a multidisciplinary approach in Bolivia. Part II. Antimalarial activity of some plants used by Mosetene indians. AB - Forty-six different species collected in the Mosetene ethnia, dwelling in the Andean Piedmont of Bolivia, were screened for antimalarial properties. Thirty three extracts were screened for antimalarial activity in vitro on Plasmodium falciparum chloroquine resistant strain (Indo), and forty-seven extracts were evaluated in vivo on the rodent malaria P. vinckei petteri 279BY. Only two plants are specifically used in combination by the Mosetene against malaria attack (Hymenachne donacifolia and Tesseria integrifolia), but they did not display any activity in vivo at 1000 mg/kg. The in vivo most active extracts were Swietenia macrophylla bark, Trema micrantha bark and Triplaris americana bark, not all of them were used for antimalarial purposes by the Mosetene. The following extracts were moderately active: Jacaratia digitata inner bark and Momordica charantia aerial part (both traditionally used as febrifuge), Kalanchoe pinnate aerial part (used in inflammatory processes), Lunania parviflora twigs and leaves, Phyllanthus acuminatus (used as piscicide), Tynanthus schumannianus fruit (used against diarrhoea), Triumfetta semitrilobata (used as febrifuge, to alleviate kidney and gynecological pain) and finally Solanum mammosum fruit (used against scabies). We present here the results of this screening, emphazing on the in vivo antimalarial activity of the selected plants. The antimalarial in vivo activity of the selected species, in relation with their traditional Mosetene use is then discussed. PMID- 10687871 TI - Evaluation of the efficacy of Lawsonia alba in the alleviation of carbon tetrachloride-induced oxidative stress. AB - The hepatoprotective activity of the 50% ethanol extract of the bark of Lawsonia alba syn. L. inermis was investigated against the carbon tetrachloride-induced oxidative stress. Pretreatment of rats with doses of 250 and 500 mg/kg of the plant extract significantly (P < 0.001) lowered serum transaminases (GOT and GPT) and LDH levels, respectively, in a dose dependent manner against the significant (P < 0.001) rise of these damage marker enzymes when challenged with CCl4 (1 ml/kg, orally). Parallel to these changes, the plant extract prevented CCl4 induced oxidative stress by significantly maintaining the levels of reduced glutathione (GSH), its metabolizing enzymes and simultaneously inhibiting the production of free radicals. Pretreatment of rats with the extract also inhibited the peroxidation of microsomal lipids in a dose-dependent manner. PMID- 10687872 TI - Ethnobotanical inventory of medicinal plants in Bulgaria. AB - The paper reports on 73 medicinal plants of 30 families, traditionally used in Bulgarian phytotherapy. Some of the plants are applied in practice for the treatment of cardiovascular, gastrointestinal, respiratory, urogenital and other disorders. The popular plants used for treatment are growing in 20 forested regions in Bulgaria. PMID- 10687873 TI - Anti-inflammatory effects of fangchinoline and tetrandrine. AB - Fangchinoline and tetrandrine are the major alkaloids from Stephania tetrandrae S. Moore which has been used traditionally for the treatment of inflammatory diseases in oriental countries including Korea. Both fangchinoline and tetrandrine showed anti-inflammatory effects on mouse ear edema induced by croton oil. In addition, the effects of fangchinoline and tetrandrine on cyclooxygenase, murine interleukin-5 (mIL-5) and human interleukin-6 (hIL-6) were examined in vitro to investigate the anti-inflammatory action mechanisms. One hundred micromolar of fangchinoline showed 35% of inhibition on cyclooxygenase, but the same concentration of tetrandrine did not show any inhibition. On the other hand, 12.5 microM of tetrandrine exhibited 95% of inhibition on mIL-5 activity, while fangchinoline did not show any effects. However, 4 microM of fangchinoline and 6 microM of tetrandrine showed 63 and 86% of inhibitions on hIL-6 activity, respectively. These results suggest that biochemical mechanisms of fangchinoline and tetrandrine on anti-inflammation are significantly different even though they are similar in chemical structure. PMID- 10687874 TI - Fetotoxicity and reproductive effects of monocrotaline in pregnant rats. AB - Four groups of 12 pregnant Wistar rats each were fed with rations containing 0, 0.01, 0.015 and 0.02% of monocrotaline (MCT) from day 6 to 21 of gestation. Liver weights of the dams from the three experimental groups were significantly lower than those from the control group. Serum levels of aspartate aminotransferase; alkaline phosphatase; lactate dehydrogenase; gamma glutamyltransferase, urea and creatinine were significantly higher in dams from MCT 0.02% group. The weights of the placenta, fetuses and fetal lungs of the 0.02% MCT group were significantly lower than those of the control group. A mild to moderate interstitial pneumonia and liver lesions were observed in dams ingesting 0.02% of MCT. These results showed the toxicity of MCT to the females that ingested 0.02% and their fetuses. Because there was no differences on the weight gains and food and water consumption of the dams it is suggested that this toxic effects in the fetuses was caused by the diffusion of MCT through the placenta. No significant differences were observed in the frequency of skeletal and visceral malformation or anomalies between the control and treated groups suggesting that MCT had no teratogenic effect. PMID- 10687875 TI - Effect of crude extracts of Erythrina americana Mill. on aggressive behavior in rats. AB - Three alkaloid fractions were obtained from seeds of Erythrina americana: free alkaloids in hexane, free alkaloids in methanol and liberated alkaloids. The pharmacological evaluation of these fractions on rats showed that, administered in a dose of 3 mg/kg, the three fractions diminished the aggressive behavior. This is comparable when diazepam is used as a control. An interaction between the cholinergic and GABAergic system could be suggested. PMID- 10687876 TI - It's time to stop using the word 'replacement'. PMID- 10687877 TI - Andropause. AB - Although, in distinction to middle aged women, in middle aged men there does not occur a sudden arrest of gonadal functions, fertility persisting until very old age, aging in men is, nevertheless, associated with an gradual decline of both endo- and exocrine testicular function. Whereas age has in fact only minimal effects on the quality of the ejaculate, endocrine function declines steadily with age and at age 75 years, mean plasma testosterone levels are only 65% of levels in young adults whereas over 25%, of these men have bioavailable testosterone levels below the lower normal limit in young adults. The interindividual variations in the plasma levels are, however, very important and a quarter of men over 75 years old, have still testosterone levels within the upper quartile of values in young men. Aging is accompanied by a series of signs and symptoms, many of which are rather similar to those observed in young hypogonadal males. The etiology of these signs and symptoms is often multifactorial, and very few correlations have been found between symptoms and plasma testosterone levels. Nevertheless, there is good evidence that the age associated decrease in testosterone levels is at least a co-determinant of these symptoms and testosterone supplementation has shown favorable effects on many of them. Side effects of this substitutive therapy are minimal when care is taken to keep plasma testosterone levels within the physiological range. Clinical prostatic carcinoma is an absolute contra-indication for testosterone supplementation. So far, there are no indications that testosterone would stimulate the evolution of as subclinical prostatic carcinoma to a clinical carcinoma but it should be recalled that so far, only a small number of elderly males received substitutive androgen treatment for longer periods in controlled studies. Hence, although side effects are generally minimal, one should, nevertheless, await the results of larger, long term, well-controlled studies before to recommend the routine testosterone substitution of elderly men. PMID- 10687878 TI - Quality of life after the menopause: a population study. AB - OBJECTIVE: To assess the impact of menopause and some sociodemographic variables on quality of life (QoL). MATERIALS AND METHODS: Four hundred and eighty-one women aged 40-59 years attending the Southern Metropolitan Health Service in Santiago de Chile were studied using the Specific Quality of Life Questionnaire for Menopause from Toronto University. RESULTS: Univariate analysis showed that menopausal women have worse QoL scores than women conserving cycles in the four areas of the questionnaire: They show a 10.6-fold higher risk for suffering vasomotor disorders affecting QoL, a 3.5-fold higher risk for psychosocial impairment, a 5.7-fold higher risk for physical disorders, and a 3.2-fold higher risk for sexual disorders (P < 0.0001). Regarding the influence of social markers (age, marital status, school years, work, number of children and sexual activity), housewives were found to have higher, worse, scores than working women in all test components (vasomotor, 3.11+/-1.90 versus 2.57+/-1.71, P < 0.003; psychosocial, 3.44+/-1.59 versus 2.92+/-1.45, P < 0.0007; physical, 3.45+/-1.36 versus 2.96+/-1.20, P < 0.0001; sexual, 3.63+/-2.23 versus 2.49+/-1.95, P < 0.0001). However, logistic regression demonstrated that the only variable found to cause a significant impairment in QoL was menopause. CONCLUSION: Menopause causes a decrease in quality of life, which is independent from age and other sociodemographic variables. PMID- 10687879 TI - Determinants of elevated blood pressure in women around menopause: results from a cross-sectional study in Italy. AB - OBJECTIVE: To analyse the determinants of high blood pressure in women around menopause. METHODS: Eligible women were consecutively identified among patients who asked for a visit of their general practitioner during the period March November 1997. A total of 22919 women aged 44-66 years (median age 55 years), were identified. Women whose mean of the second and third of the three measures of diastolic blood pressure values performed during interview was > 90 mm of mercury and/or reporting any type of current pharmacological treatment for elevated blood pressure were considered hypertensive. RESULTS: In comparison with women aged 40-50 years, the multivariate odds ratio (OR) of elevated blood pressure were 1.4 in women aged 51-55, 2.0 in those aged 56-60 and 2.7 in those aged > or = 61. In comparison with women with a body mass index (kg m(-2)) < 25, the OR of elevated blood pressure was 1.7 and 2.7, respectively, for women with a BMI of 25 28 and > or = 29. In comparison with women reporting a low level of physical activity, the OR of elevated blood pressure were 0.9 (95%, confidence interval, CI 0.7-1.0) and 0.7 (95% CI 0.4-0.9), respectively, for those reporting an intermediate or high level of activity. In comparison with premenopausal women, the OR of elevated blood pressure was, after taking into account the confounding effect of age, 1.6 (95% CI 1.0-1.4) in post menopausal ones. The OR of elevated blood pressure was 0.8 (95% CI 0.7-0.9), for current users of hormone replacement therapy (HRT), but no clear association emerged with duration of HRT pressure. CONCLUSIONS: This study suggests that, after taking into account the effect of age, post-menopausal women are at higher risk of the condition, and current HRT use decreases the risk. Other main determinants of risk of elevated blood pressure were overweight and low physical activity. PMID- 10687880 TI - Does apolipoprotein E genotype relate to BMD and bone markers in postmenopausal women? AB - OBJECTIVES: Bone mineral density (BMD) and development of osteoporosis are partly determined by genetic factors. The associations between one of suggested candidate, apolipoprotein E (apo E) genotype to bone mineral density (BMD) and bone biochemical markers was studied in 464 subjects recruited from a population based group of early postmenopausal women (n = 13100). Additionally, the influence of apo E genotype on BMD changes during a 5-year follow-up with or without hormone replacement therapy (HRT) was investigated. METHODS: Participants were randomized into two treatment groups: HRT group: Sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate with or without vitamin D3, 100-300 IU/day + calcium lactate, 500 mg/day (n = 232), and the non-HRT group: Calcium lactate, 500 mg/day alone or in combination with vitamin D3, 100-300 IU/day (n = 232). BMD was measured from the lumbar spine and proximal femur at baseline and after 5 years of treatment (n = 352). In a subgroup (n = 59), the serum concentrations of bone biochemical markers (intact osteocalcin (OC), bone specific alkaline phosphatase (BAP) and type I collagen carboxy-terminal telopeptide (ICTP)) were measured at baseline and after 1 year of follow-up. RESULTS: At baseline, the BMDs were similar between the five apo E genotype groups (2/3, 2/4, 3/3, 3/4, 4/4). No significant differences in lumbar or femoral neck BMDs of women with the apo E4 allele were found compared with those without it. There was a statistically significant difference in 5-year BMD changes between the HRT and non-HRT groups. After 5 years, the BMD of the femoral neck had remained constant and the mean lumbar spine BMD had increased by 1.5% in the HRT group, whereas both BMDs had decreased by 4-5% in the non-HRT group. However, the apo E genotype did not modify the changes in BMD in either group. Additionally, the baseline concentrations of bone metabolic markers and their 1 year changes showed no genotype-related associations. CONCLUSIONS: The results of our population-based study indicate that apo E genotype does not modify lumbar or femoral neck BMDs or serum bone biochemical markers or their response to HRT in early postmenopausal Caucasian women. PMID- 10687881 TI - Hypersecretion of ovarian androgens may be gonadotrophin dependent many years after menopause. AB - BACKGROUND: In fertile women both adrenals and ovaries contribute to androgen production, whereas after the menopause the ovarian contribution normally decreases. OBJECTIVE: The objective of this case study was to assess whether ovarian androgen secretion was responsive to decreased gonadotrophin stimulation and whether gonadotrophins were sensitive to negative feedback from sex steroids many years after the menopause. METHODS: In this uncontrolled case study a 72 years old slightly overweight woman with noninsulin-dependent diabetes mellitus presented with hirsuitism and elevated serum testosterone concentrations. The woman was reluctant to have an oophorectomy, and received an oral estradiol/progestagene preparation. Serum testosterone and gonadotrophin concentrations were measured before and after steroid hormone therapy. RESULTS: Serum gonadotrophin concentrations decreased and testosterone levels returned to normal during therapy. When the hormone therapy was stopped for 1 month the high testosterone concentrations returned, but were again normalized when the hormone therapy was reinitiated. CONCLUSION: The ovaries of this woman were apparently still responsive to pituitary stimulation and her hypothalamic pituitary ovarian feed-back system still seemed to be working after 70 years of age. PMID- 10687882 TI - Efficacy and tolerability of a new estradiol delivering matrix patch (Estraderm MX) in postmenopausal women. AB - OBJECTIVE: To examine the efficacy and tolerability of a new matrix patch delivering estradiol (E2 Matrix) at doses of 0.05 and 0.10 mg per day (Estraderm MX 50, 100) in the treatment of moderate to severe postmenopausal symptoms. METHODS: A total of 254 postmenopausal women were randomized to receive treatment with E2 Matrix 0.10 mg (N = 86), E2 Matrix 0.05 mg (N = 82), or placebo (N = 86) in a double-blind, double-dummy fashion for a period of 12 weeks continuously. Patches were applied twice weekly to the buttocks with each patient wearing two patches at all times. The primary efficacy criterion was the difference from baseline of the mean number of moderate to severe hot flushes per 24 h during the last 2 weeks of treatment. Other efficacy variables included reduction in hot flushes at 4 and 8 weeks, reduction in daytime flushing and night sweats, and Kupperman Index at 4, 8, and 12 weeks. RESULTS: E2 Matrix 0.10 and 0.05 mg were both significantly superior to placebo in reducing hot flushes per 24 h after 4, 8, and 12 weeks of treatment (P < 0.001). Also, for all other efficacy parameters studied, both dosage strengths of E2 Matrix were statistically significantly superior to placebo at all time points (P < 0.001). Local tolerability was good in both groups. A slight increase in estrogen related adverse effects (breast tenderness, leukorrhoea) was seen with the 0.10 mg patch. Adhesion of patches and compliance were good. Overall systemic tolerability was good in both treated groups. However, a 4.8% overall incidence of endometrial hyperplasia was observed in patients with an intact uterus. CONCLUSIONS: This new matrix patch offers an effective and well tolerated dosage form for delivery of 0.05 and 0.1 mg estradiol per day. It may be particularly suitable for those women who experience local sensitivity to alcohol-containing systems. In light of the observed hyperplasia after treatment in five patients, estrogen therapy should as yet be supplemented monthly with a progestogen in women with an intact uterus. PMID- 10687883 TI - Bioavailability of estradiol from two matrix transdermal delivery systems: Menorest and Climara. AB - OBJECTIVES: To compare two estradiol transdermal matrix systems with regard to bioavailability, pharmacokinetics and tolerability. METHODS: A single centre, open, randomized, comparative cross-over study in 20 healthy postmenopausal women. Menorest with 3 or 4 days of suggested use and Climara with 7 days of suggested use (both 50 microg/24 h) were compared at steady state. Two 14-day treatment periods were separated by a 4 week washout. Plasma levels of estradiol were monitored during the second week of each treatment. Tolerability was assessed by open questions and inspection of the application site. RESULTS: There were no differences between the two treatments with regards to AUC, Cmax, Cmin, Caverage or fluctuations of plasma estradiol. Tmax was significantly shorter for Menorest than Climara. Cmax and Cmin were significantly higher for the second Menorest patch during the monitoring period compared to the first. All local reactions were mild and there were three cases of erythema with Menorest and a total of 21 skin reactions in 15 subjects with Climara. Systemic tolerability was similar between treatments with eight estrogen-related adverse events in eight subjects (period pains, uterine bleeding, mastodynia, headache and vaginal discharge) with Menorest and 13 events in ten subjects with Climara. CONCLUSIONS: The bioavailability of estradiol from the two matrix transdermal delivery systems Menorest and Climara was similar, but the products were not bioequivalent because Tmax was significantly shorter for Menorest than for Climara. Tolerability of treatment was good for both patches but with a higher number of local reactions and estrogen related adverse events for Climara. PMID- 10687884 TI - Characterization of hot flashes reported by healthy postmenopausal women receiving raloxifene or placebo during osteoporosis prevention trials. AB - OBJECTIVE: Raloxifene, a selective estrogen receptor modulator, is estrogen-like in the skeleton and cardiovascular system and antiestrogenic in reproductive tissues. In contrast to estrogens, raloxifene is not indicated for the treatment of hot flashes. This study was designed to examine the characteristics of hot flashes among healthy postmenopausal women participating in osteoporosis prevention trials who were receiving raloxifene or placebo. METHODS: Adverse event data from three randomized, double-blind trials (N = 876) comparing raloxifene 60 mg/day with placebo for 30 months were integrated and analyzed. Two of the three trials (one European, two North American) were identically designed and were open to healthy postmenopausal women ages 45 through 60 without regard to prior hysterectomy. The third trial was multinational, was open to women ages 40 through 60, and all enrollees had prior hysterectomy at baseline. Women were questioned in general terms about the occurrence of adverse events at 3-6-month intervals. Treatment-emergent adverse events pertaining to hot flashes were included in the current study. RESULTS: At baseline, 12% of women randomly assigned to placebo and 13% assigned to raloxifene reported prevalent hot flashes. After 30 months, the cumulative incidence of hot flashes was 21% for placebo and 28% for raloxifene (P = 0.022), with the difference in incidence rate confined to the first 6 months of therapy. There was no difference between placebo and raloxifene in reported maximum severity of or early discontinuations as a result of hot flashes (< or = 3% per group for both outcomes). Among women whose hot flashes had stopped completely during the 30-month study period, the median total duration of the event prior to becoming symptom-free was 246 days for placebo and 205 days for raloxifene. Among all women reporting a hot flash, the extrapolated total duration of hot flashes was the same for women treated with either raloxifene or placebo. No subgroup-by-therapy interactions were detected. Multivariable regression analysis revealed several factors that were independently weakly predictive of hot flashes. CONCLUSIONS: Raloxifene slightly affects the incidence but not the natural history of hot flashes in healthy postmenopausal women seeking prevention therapy. PMID- 10687885 TI - The effect of hormone replacement therapy on metabolism of lipoprotein remnants in postmenopausal women. AB - OBJECTIVE: The measurement of remnant-like particles reflects chylomicron and very low density lipoprotein remnants which are most likely atherogenic particles. We investigated the effects of menopausal status and postmenopausal hormone replacement on metabolism of remnant lipoprotein-cholesterol. METHODS: We measured remnant lipoprotein-cholesterol by an immunoseparation assay in 20 premenopausal, 40 postmenopausal, and 30 bilaterally oophorectomized women. Of 70 postmenopausal subjects, 21 surgically menopausal women (with total hysterectomy) were started on hormone replacement with conjugated equine estrogen, 0.625 mg/day, and 36 naturally postmenopausal women were begun on a combination of conjugated equine estrogen 0.625 mg/day, plus medroxyprogesterone acetate, 2.5 mg/day. Plasma levels of remnant lipoprotein-cholesterol and other common lipids were measured after 6 and 12 months of treatment. RESULTS: Plasma remnant lipoprotein-cholesterol levels in postmenopausal and surgically menopausal women were significantly higher than in premenopausal women (P < 0.005). Plasma total and low-density lipoprotein cholesterol levels decreased and high-density lipoprotein cholesterol increased significantly (P < 0.01) in both treatment groups, respectively. Plasma triglyceride levels were not changed by treatment; however, remnant lipoprotein-cholesterol levels decreased in both treatment groups (estrogen group; P = 0.07, estrogen-progestin group; P < 0.05). No side effects of therapy were consistently reported. CONCLUSIONS: We confirmed that remnant lipoprotein-cholesterol increases after menopause. Hormone replacement therapy improves disordered lipoprotein metabolism and exerts a favorable effect on lipoprotein remnant metabolism in postmenopausal women. PMID- 10687886 TI - Effects of combined sex hormone replacement therapy on small artery biomechanics in pharmacologically ovariectomized rats. AB - OBJECTIVES: The purpose of this study was to determine the effects of long-term combined sexual hormone replacement therapy on the biomechanical properties of the small artery wall in castrated female rats. METHODS: 30 non-pregnant mature female Sprague-Dawley rats were pharmacologically ovariectomized with 750 microg/kg triptorelin im. every 4th week. Ten of them received combined hormone replacement in form of 15 mg/kg medroxyprogesterone acetate (MPA) im. every 2 weeks and 450 microg/kg estradiol propionate im. once a week. Ten castrated animals received MPA only. Ten control, castrated animals were given the vehicles of these steroids. Ten other animals were kept parallelly, receiving the vehicles of all drugs (control animals). After 12 weeks of treatment cylindrical segments of the saphenous artery were isolated and cannulated at both ends and subjected to in vitro microarteriographic test. Pressure diameter curves, in the range of 0 200 mmHg, were recorded from segments in normal Krebs-Ringer (nKR) solution, in contraction with norepinephrine (1.6 x 10(-5) M), and then in relaxation with papaverine (2.8 x 10(-5) M). Biomechanical parameters were calculated based on the pressure diameter curves. RESULTS: Combined hormone replacement therapy significantly increased the passive diameter of small arteries, as compared to those from ovariectomized animals without hormone replacement. MPA monotherapy did not alter the vessel diameter, the inner radii at 100 mmHg intraluminal pressure were, 300+/-9 microm in the control castrated, 340+/-7 microm in the estradiol + MPA replaced and 306+/-8 microm in the MPA treated groups (P < 0.05 between the control castrated and the combined treatment groups). The vascular reactivity to norepinephrine or papaverine was not changed significantly either by combined hormone replacement or by MPA monotherapy when compared with ovariectomized controls. No significant alterations were found in wall thickness and distensibility. CONCLUSIONS: These results suggest that chronic medroxyprogesterone pretreatment does not influence the geometric, elastic and contractile properties of small arteries in castrated female rats. The combination of MPA + estradiol increased the morphological lumen: the morphological vasodilatation induced by estrogen, described earlier, was not affected by the addition of this progestin to the regimen. PMID- 10687887 TI - Vegan proteins may reduce risk of cancer, obesity, and cardiovascular disease by promoting increased glucagon activity. AB - Amino acids modulate the secretion of both insulin and glucagon; the composition of dietary protein therefore has the potential to influence the balance of glucagon and insulin activity. Soy protein, as well as many other vegan proteins, are higher in non-essential amino acids than most animal-derived food proteins, and as a result should preferentially favor glucagon production. Acting on hepatocytes, glucagon promotes (and insulin inhibits) cAMP-dependent mechanisms that down-regulate lipogenic enzymes and cholesterol synthesis, while up regulating hepatic LDL receptors and production of the IGF-I antagonist IGFBP-1. The insulin-sensitizing properties of many vegan diets--high in fiber, low in saturated fat--should amplify these effects by down-regulating insulin secretion. Additionally, the relatively low essential amino acid content of some vegan diets may decrease hepatic IGF-I synthesis. Thus, diets featuring vegan proteins can be expected to lower elevated serum lipid levels, promote weight loss, and decrease circulating IGF-I activity. The latter effect should impede cancer induction (as is seen in animal studies with soy protein), lessen neutrophil-mediated inflammatory damage, and slow growth and maturation in children. In fact, vegans tend to have low serum lipids, lean physiques, shorter stature, later puberty, and decreased risk for certain prominent 'Western' cancers; a vegan diet has documented clinical efficacy in rheumatoid arthritis. Low-fat vegan diets may be especially protective in regard to cancers linked to insulin resistance--namely, breast and colon cancer--as well as prostate cancer; conversely, the high IGF-I activity associated with heavy ingestion of animal products may be largely responsible for the epidemic of 'Western' cancers in wealthy societies. Increased phytochemical intake is also likely to contribute to the reduction of cancer risk in vegans. Regression of coronary stenoses has been documented during low-fat vegan diets coupled with exercise training; such regimens also tend to markedly improve diabetic control and lower elevated blood pressure. Risk of many other degenerative disorders may be decreased in vegans, although reduced growth factor activity may be responsible for an increased risk of hemorrhagic stroke. By altering the glucagon/insulin balance, it is conceivable that supplemental intakes of key non-essential amino acids could enable omnivores to enjoy some of the health advantages of a vegan diet. An unnecessarily high intake of essential amino acids--either in the absolute sense or relative to total dietary protein- may prove to be as grave a risk factor for 'Western' degenerative diseases as is excessive fat intake. PMID- 10687888 TI - Incorporation of liposome-encapsulated amphotericin in artificial/prosthetic cardiac valves for therapy and prevention of fungal endocarditis. AB - Fungal endocarditis is a highly fatal condition. Fungal prosthetic valve endocarditis is an uncommon but serious infection. Patients require surgery and long-term antifungal therapy. However, recurrence is quite common and eradication of infection is difficult. Liposomal amphotericin is considered to be better than the conventional preparation. It is hypothesized that incorporation of liposome encapsulated amphotericin inside artificial/prosthetic cardiac valves would result in better tissue concentration of the drug at site of infection and probable prevention of recurrence. The cost of making the same might be considerable but would likely be cost-effective in the long run due to lowered mortality and prevention of recurrence. PMID- 10687889 TI - Prevention of progression in chronic myeloid leukemia by altering DNA methylation with a pyridoxine analogue. AB - Altered DNA methylation is one mechanism by which progression might occur in tumors. Progression in chronic myeloid leukemia (CML) was initially thought to be due to mutations. However, altered DNA methylation might be responsible for the same. An agent whose probable target is DNA methylation has been shown to improve hematopoiesis in CML. Pyridoxine is involved in methylation reactions. At present, the link between dietary factors like pyridoxine and cancer is somewhat tenuous. It is hypothesized that a suitable pyridoxine analogue might, by blocking DNA methylation, prevent progression in CML. PMID- 10687890 TI - Anesthesia is a risk factor for drug and alcohol craving and relapse in ex abusers. AB - Relapse to cocaine, heroin, and alcohol is a common occurrence in ex-abusers of these substances. Although there are many potential causes for relapse, we postulate that one cause in some people may be re-exposure in an anesthesia setting to a drug similar to the formerly abused drug. We hypothesize, for example, that opioids given during and after surgery may reinstate craving for, and initiate subsequent seeking of, heroin in former abusers. There are a substantial number of animal studies and some human studies documenting a reinstatement phenomenon in which an experimenter-administered psychoactive drug can precipitate drug-seeking behavior in 'abstinent' animals and humans. There is concern amongst health professionals and patients alike on this issue, and we discuss possible avenues of research, both preclinical and clinical, to explore the validity of our hypothesis. PMID- 10687891 TI - Non-linear model of cancer growth and metastasis: a limiting nutrient as a major determinant of tumor shape and diffusion. AB - A new approach for modelling the spatio-temporal evolution of tumors is presented. To test its validity, a very basic model is considered, which, in spite of its simplicity, is capable of generating a multiplicity of morphologies and growth and migration rates. From an in-vivo scenario of basic life processes, cancer cell proliferation is described as a competition for basic nutrients. The chosen mathematical treatment and simulation techniques permit a direct implementation of the local nonlinear couplings existing between the various cell populations and the free and bound nutrient concentration. A discussion of the results and proposed improvements and applications of the model is also presented. PMID- 10687892 TI - Directed evolution of light-activated drugs. AB - Photodynamic therapy is a new technology that uses light-activated molecules to target drug action to diseased areas while sparing healthy tissue. Unfortunately, the present arsenal of photosensitive molecules is limited in both number and scope. We hypothesize that new photosensitive molecules could be developed using directed molecules evolution. This paper outlines a possible selection mechanism to evolve molecules activated by a desired wavelength of light. PMID- 10687893 TI - The oxidative stress hypothesis of atherosclerosis: cause or product? AB - The oxidative stress hypothesis postulates that endogenous free radicals of unknown origin, possibly derived from mural cells, oxidize low density lipoproteins and that oxidation products are allegedly responsible for initiation and progression of atherosclerosis. The thesis fails to explain its topography, site specific severity and the iatrogenic and experimental hemodynamic induction of atherosclerosis under conditions complying with the logic of Koch's postulates. Free radicals are generated by biomechanical scission of macromolecules and polymers, the biophysical mechanism underlying bioengineering fatigue in atherogenesis with oxidative damage a secondary, contributory factor to mural pathology. The plentiful supply of antioxidants negates oxidative stress as the dominant factor in atherogenesis. PMID- 10687894 TI - Role of brain organization in the pathogenesis of physical disease. AB - While genetic and environmental factors are known to make substantial contributions to the pathogenesis of physical disease, the role of the brain in these processes is largely unknown. It is hypothesized that the manner in which the brain is functionally organized is an integral factor in physical health outcomes, both positive and negative. It is further hypothesized that changes in certain patterns of the functional organization of the brain play a significant role in the pathogenesis of physical disease, mediating between an individual's genetic endowment, the environment, and other relevant brain systems to initiate, modulate and/or maintain these disorders. There are many currently available treatment modalities which have the capacity to change the pattern of functional brain organization. Such interventions have the potential to become valuable aids in both the treatment and prevention of physical disease. PMID- 10687895 TI - One explanation of Nef gene behavior. AB - The Nef (negative factor) gene of primate retroviruses may serve an important evolutionary function. Selection pressures in the natural world, may at times, demand that retroviral infection not cause disease in a newly entered host species. Genetic alterations in the Nef gene may function to permit retroviral speciation by lowering retroviral replication rate. Slow viral replication may allow small numbers of the newly infected host species to avoid the effects of retroviral pathology. PMID- 10687896 TI - A biophysical basis of enhanced interstitial fluid pressure in tumors. AB - It is widely accepted that enhanced interstitial fluid pressure (IFP) in tumors is a major obstacle against delivery of therapeutic agents. On the other hand, the origin of enhanced IFP remains controversial. Here, the Van't Hoff equation is applied to examine how glucose breakdown to CO2 and lactate in tumor cells may affect intracellular osmotic pressure. According to the equation, it is found that production of CO2 from glucose lowers osmotic pressure inside cells, while glycolytic production of lactate generates significant increases. Crucial to a net enhancement of pressure in cells is the Warburg ratio, the ratio of the fraction of glucose transformed to lactate divided by the fraction of glucose metabolized to CO2: if (and only if) the ratio is higher than 1.0, there is a resulting increase in intracellular osmotic pressure. Under fully anaerobic glycolysis, the enhancement of intracellular pressure is maximal, namely 19.3 mmHg per mM of glucose metabolized to lactate (Van't Hoff equation). Cells are then biological pressure pumps driven by glycolytic production of lactate, causing IFP to raise. It is proposed that a regulatory feedback loop prevents IFP to raise above microvascular pressure (MVP). Accordingly, enhanced IFP in tumors is the result of high rates of tumor glycolysis, and enhancement of IFP is limited by MVP. It is thus concluded that a high rate of glycolytic production of lactate in tumor cells ultimately prevents both access of therapeutic agents to the malignant cells and immunological surveillance, and that it indirectly drives outward currents of interstitial fluid, thereby propelling both the process of tumor infiltration of surrounding structures and metastatic spread, depending on deformability and proteolytic capacity of the malignant cells. PMID- 10687897 TI - Could growth retardation in cystic fibrosis be partly due to deficient steroid and thyroid hormonogenesis? AB - Cystic fibrosis (CF) mainly affects Caucasians of northwestern-European ancestry with severe morbidity. The individuals are malnourished and growth retarded. The latter is thought to be the consequence of delayed maturation of the hypothalamic pituitary-gonadal axis due to malnourishment. However, there is evidence that steroid and thyroid hormone syntheses may be impaired in CF. Thyrotropin stimulates the uptake and efflux of iodide before the halide is incorporated into thyroid hormones and it is becoming apparent that gonadotropins likewise mobilise chloride ions in Leydig cells prior to steroidogenesis. Since the primary defect causing CF is the mutated cystic fibrosis transmembrane conductance regulator (CFTR)--a chloride channel residing on the apical membrane of wet epithelia, the growth retardation in CF may in part be due to deficient hormone syntheses. The latter may involve CFTR or may be the halide channel activated by glycoprotein hormones prior to hormonogenesis. PMID- 10687898 TI - Occlusions of epicardial arteries might not directly induce symptoms in ischemic heart disease. AB - It is accepted that primary occlusions of epicardial arteries by thromboses, stenotic coronary artery disease (CAD), and spasm directly induce symptoms in ischemic heart disease (IHD). Because of this acceptance, there has been little interest in alternate mechanisms for IHD--as the spasm of resistance vessel (S RV) concept of IHD, which asserts that S-RV directly induces symptoms in IHD. To stimulate interest in the S-RV concept, evidence against the primacy of occlusions of epicardial arteries was presented, as well as evidence for this position to provide a balanced discussion; while the evidence was mixed, overall findings appeared to weigh significantly against the primacy of occlusions of epicardial arteries. Also, the S-RV concept was discussed; the discussion included presenting the theory's explanations for events in epicardial arteries, with the aim of demonstrating that the concept provides more consistent explanations than the standard position. It is suggested that there is sufficient information to warrant renewed consideration of the S-RV concept. PMID- 10687899 TI - Digitoxin is a potential anticancer agent for several types of cancer. AB - The ability of digitalis to block cell proliferation has been well established for some time. Recently, digitalis in non-toxic concentrations has been showed to induce apoptosis in different malignant cell lines. In light of the pivotal role of apoptosis in cancer development and progression and this new experimental finding concerning digitalis, it seems probable that the apoptosis-inducing capability is explained by mechanisms other than just Na+/K+ ATPase inhibition. In this article, features of the cardiac glycosides which make them interesting to evaluate further as potential anticancer drugs are discussed. Some new data concerning inhibition and apoptosis in three human glioblastoma cell lines by digitoxin are also presented. PMID- 10687900 TI - Prognostic factors and relative risk for survival in N1-3 oral squamous cell carcinoma: a multivariate analysis using Cox's hazard model. AB - The records of 136 patients with N1-3 oral squamous cell carcinoma treated by surgery were investigated retrospectively, with the aim of finding out which factors were predictive of survival on multivariate analysis. Four independent factors significantly influenced survival in the following order: pN stage; T stage; histological grade; and N stage. The most significant was pN stage, the five-year survival for patients with pN0 being 91% and for patients with pN1-3 41%. A further study was carried out on the 80 patients with pN1-3 to find out their prognostic factors for survival and the independent factors identified by multivariate analysis were T stage and presence or absence of extracapsular spread to metastatic lymph nodes. PMID- 10687901 TI - A simple method of plate fixation of fractures of the frontal bone. AB - We present a simple method to reduce and align depressed fractures of the frontal sinus or cranial vault before rigid fixation using microplates. PMID- 10687902 TI - Mandibular third molars: oral surgeons' assessment of the indications for removal. AB - The aim was to examine oral surgeons' assessment of the indications for removal of mandibular third molars. Questionnaires were distributed to seven oral and maxillofacial surgery clinics. The oral surgeons were asked to record whether or not there was associated disease. Three other factors were recorded: patient's age, and angular position and extent of eruption of the molars. The strength of the indication for removal was rated on a visual analogue scale (VAS) where 0= weakest and 100= strongest indication for removal. The results were based on data from 666 molars: 118 (18%) had no disease, 465 (70%) had one associated disease, 77 (11%) had two and 6 (1%) had three. The indication for removal as expressed by the mean VAS for molars with no disease was assessed to be weaker (P<0.05) than that for molars with one, two, or three diseases. The only factor that influenced the indication for removal in molars with no disease was the patient's age. PMID- 10687903 TI - Association between paroxysmal trigeminal neuralgia and atypical facial pain. AB - Paroxysmal trigeminal neuralgia and atypical facial pain are both fairly common conditions that produce pain in the face of different character. Trigeminal neuralgia is sharp and shooting, brought on by facial movement, change of temperature and by touching the face at a specific point (the trigger point). Atypical facial pain is dull and unrelenting and its site is ill-defined. Trigeminal neuralgia is generally more common in older people, and affects women slightly more than men, and atypical facial pain generally affects younger people, with women predominating. The pains should never be confused. We have noticed that many patients with trigeminal neuralgia have additional symptoms of atypical facial pain and so we reviewed the records of the Pain Relief Unit retrospectively. Of the 83 patients identified with trigeminal neuralgia where records were adequate, 35 (42%) also had atypical facial pain. Five of these had developed it before the onset of trigeminal neuralgia and could be examples of pretrigeminal neuralgia. There were eight patients in the series with multiple sclerosis, of whom two also had atypical facial pain. There seemed to be no relationship between the development of atypical facial pain and the interventions used to treat trigeminal neuralgia. It is important that both conditions are identified and treated individually. PMID- 10687904 TI - Life-threatening haemorrhage after elevation of a fractured zygoma. AB - A 21-year-old man presented with a fractured left zygoma after an alleged assault. The fracture was elevated four days later, at which time he had a brisk left-sided epistaxis. Recovery was uneventful except for a haematoma that was drained a month later. Two weeks after this, he was admitted after having collapsed. He was shocked and bleeding profusely from his nose. He had a further major bleed in hospital and this was treated by tying off the left external carotid artery. He has made an uneventful recovery and investigations have shown no bleeding diathesis. PMID- 10687905 TI - Alveolar soft part sarcoma of the tongue. AB - We report a case of alveolar soft part sarcoma--a rare malignancy that presented as a swelling at the base of the tongue in a 5-year-old child. Only about one quarter of the few reported cases arise within the head and neck, the tongue and orbit being the favoured sites. PMID- 10687906 TI - Temporomandibular joint symptoms and disc displacement in patients with mandibular prognathism. AB - Our objective was to find out the incidence of signs and symptoms of temporomandibular joint (TMJ) and disc displacement in patients with mandibular prognathism. Fifty-one patients were examined clinically and by axial computed tomography(CT). The incidence of TMJ signs and symptoms was 6/25(24%) in patients with simple mandibular prognathism and 12/26(46%) in patients with mandibular prognathism and asymmetry. No discs were displaced in patients with simple mandibular prognathism, but 15(58%) of the patients with mandibular prognathism and asymmetry had displaced discs. There was no association between signs and symptoms of TMJ and disc displacement. Patients with mild protrusion and severe asymmetry of the mandible had a high incidence of disc displacement, which interestingly was on the deviated side in 14 of the 15 patients affected. We conclude that skeletal morphology may have a role in the development of TMJ disorders but the mechanism is obscure. PMID- 10687907 TI - The value of magnetic resonance imaging in the diagnosis of mandibular osteomyelitis. AB - Our aim was to evaluate the accuracy of magnetic resonance imaging (MRI) and bone scintigraphy in the diagnosis of mandibular osteomyelitis. Twenty patients with mandibular osteomyelitis were prospectively investigated by conventional radiography, bone scintigrams and MRI. All diagnoses were verified either by surgery or by the clinical course. There was no significant difference between bone scintigraphy and MRI in the detection of osteomyelitis or the assessment of its extent. MRI was significantly better than scintigraphy at detecting the presence and assessing the extent of extraosseous inflammation. We always use MRI to diagnose osteomyelitis. For long-term follow-up of patients with mandibular osteomyelitis, we recommend MRI and bone scintigraphy. PMID- 10687908 TI - Upper buccal sulcus approach to management of fractures of the zygomatic complex: a retrospective study of 50 cases. AB - A retrospective study was conducted of 50 consecutive cases of fractures of the zygomatic complex reduced by the upper buccal sulcus approach. All were treated successfully with simple elevation (n=38), elevation with intraoral plating at the zygomatic buttress (n=8), or extraoral placement of bone plates (n=4). In no case was the approach deemed unsuitable, or abandoned in favour of another technique. There was minimal morbidity (one case each of mild diplopia, trismus, and swelling, all of which settled spontaneously). The upper buccal sulcus approach is a safe, rapid and effective technique for the reduction of zygomatic body and arch fractures. PMID- 10687909 TI - Ostrich eggshell as a bone substitute: a preliminary report of its biological behaviour in animals--a possibility in facial reconstructive surgery. AB - The aim of this study was to assess the biological behaviour of an implant of ostrich eggshell in various animal models of facial bone reconstruction. The implant was first bioassayed in a rat muscle pouch (n=10), and then tested as an interpositional graft in rat (n=10) and rabbit (n=5) cranial defects. It was finally used as an onlay graft on rabbit mandibles (n=5). Animals were killed after two months in the bioassay, three months in the interpositional model, and six months in the onlay model. The specimens were studied by contact radiography and standard histological techniques. All animals showed normal wound-healing. In the bioassay, the implants produced only a minimal inflammatory reaction. In the interpositional model, the implants maintained a good contour, but there was no sign of graft-remodelling. In the onlay model, the grafts were stable and partly osteointegrated. The onlay graft model gave the most promising results. Because ostrich eggshell is inexpensive and has good mechanical properties, it deserves further study. Long-term studies will clarify its possible role in maxillofacial surgery. PMID- 10687910 TI - Comparison of two techniques of patient-controlled sedation with midazolam. AB - OBJECTIVE: To compare patient-controlled sedation with 1-mg increments of midazolam at 1-min intervals with 0.1-mg increments of midazolam without a lock out interval. DESIGN: Randomized cross over study. SUBJECTS: 32 patients aged 17 35 years having third molars removed. RESULTS: Doses of midazolam obtained, degree of sedation and operating conditions were similar in the two groups. The demands far exceeded the increments actually received by patients obtaining 0.1 mg increments. Some were extremely sedated with both techniques. CONCLUSIONS: In this age group, there were no significant advantages or disadvantages of one technique over the other. Patients obtained the degree of sedation they required to undergo the operation by pressing the button independently of the dose or incremental interval. So-called 'true' patient-controlled sedation is a misnomer. The cut-off interval proved to be an extremely important safety feature. PMID- 10687911 TI - Aberrations of erbB-1 and erbB-2 oncogenes in non-dysplastic leukoplakias of the oral cavity. AB - The purpose of this study was to analyse erbB-1 and erbB-2 oncogenes in non dysplastic oral leukoplakia to see if we could pinpoint the first steps towards dysplasia and possible carcinogenesis. Fresh biopsy specimens of leukoplakia in 13 patients with no history of oral cancer were examined using the competitive differential polymerase chain reaction. The mean gene copy numbers of erbB-1 and erbB-2 were calculated from the formula to compare the absolute quantities of reference gene and oncogene from 24 patients who did not have leukoplakia. Healthy mucosa was taken as controls. In eight patients with leukoplakias, the results indicated aberrations of the erbB-1 oncogene, and two patients had gene dosage changes of erbB-2. There were no signs of deletions or amplifications in the controls. These results suggest that aberrations of erbB-1 and erbB-2 are additional markers in premalignant oral lesions at the beginning of the carcinogenic process, and that genetic alterations in histologically non dysplastic premalignant oral lesions are common. PMID- 10687912 TI - Distraction implants--a new possibility for augmentative treatment of the edentulous atrophic mandible: case report. PMID- 10687913 TI - A cephalometric study of the relationship between the level of velopharyngeal closure and the palatal plane in patients with repaired cleft palate and controls without clefts. AB - To find out whether the palatal plane is a useful indicator for evaluating the level of velopharyngeal closure, we did a cross-sectional study from early childhood to puberty of the vertical relationship between the palatal plane and the level of velarpharyngeal contact during velopharyngeal functioning in 61 patients with repaired cleft palate (unilateral cleft lip and palate = cleft group) and 82 controls without clefts (control group). Measurements on the vertical dimension were derived from a coordinate system and landmarks on lateral cephalograms, and the significance of differences in measurements was analysed using Student's t-test. Changes in the points of velarpharyngeal contact in relation to the palatal plane with growth showed a consistent tendency though differed between the two groups. In the control group, the PPW (point where palatal plane extension intersects the posterior pharyngeal wall) was maintained at a level that did not differ significantly from the level of midpoint of velarpharyngeal contact during phonation of /a/, and was maintained at a level that did not differ significantly from the level of the inferior point of velarpharyngeal contact. In the cleft group, however, it was maintained at a level that was slightly higher than the superior point of velarpharyngeal contact both during phonation of /a/ and during blowing. These results suggest that the palatal plane is useful as an indicator for evaluating the level of velopharyngeal closure. PMID- 10687914 TI - Effects of surgical reduction of the tongue. AB - There are still no objective diagnostic criteria for macroglossia. The aims of this study were to examine the effect of reduction of the tongue on its position, and to suggest a standard measurement point for the objective diagnosis of macroglossia. Twenty-four patients were studied. Lateral cephalometric radiographs were taken with the tongue in the rest position, and preoperative and postoperative measurements were made on cephalometric lateral roentgenograms from all 14 patients with macroglossia and the 10 control subjects. The results suggested that measurement of length of the tongue, and the tongue area: oral cavity ratio on lateral cephalometric radiographs are useful in the more accurate diagnosis of macroglossia. PMID- 10687915 TI - Device for collecting bone from the ilium for bone transplantation in cleft jaw. PMID- 10687916 TI - Specialist practitioner' in surgical dentistry: preliminary report on a pilot scheme at East Surrey Hospital, Redhill, UK. AB - This continuing pilot scheme was designed to assess whether minor oral operations could be done by a 'specialist practitioner' in surgical dentistry in hospital. The preliminary results indicate that patients benefit from the improved facilities and expertise that are available in the hospital, that the provision of treatment within the hospital is at a sufficiently 'local' level to meet their requirements, and that this increased quality of service can be provided at no greater cost to the NHS than treating them in a dental surgery. A large amount of the dentoalveolar surgery done (such as removal of third molars) is regarded as routine, but 'routine' is often mistakenly thought to mean 'simple' or 'easy'. Dentoalveolar surgery not only demands the highest quality of care and expertise but it also requires the necessary immediate support if medical or surgical complications arise. PMID- 10687917 TI - Need for intensive care after operations for head and neck cancer surgery. AB - We compared the postoperative morbidity of 44 patients who had had major head and neck oncological resections and who were nursed postoperatively on a general ward with that of 33 who were nursed on an intensive care unit at North Manchester General Hospital and Withington Hospital, South Manchester, respectively. There was no difference in the general morbidity (9/44, 20% compared with 9/33, 27%, 95%, CI of difference -0.26 to 0.13). We conclude that it is safe to nurse the patients on a general ward provided that certain conditions are fulfilled. PMID- 10687918 TI - Re: Bos et al. A randomised controlled trial of the management of mandibular condyle fractures. PMID- 10687919 TI - Re: Edwards et al. Choice of anaesthetic and health care facility for third molar surgery. PMID- 10687920 TI - Wisdom tooth removal and lingual nerve damage. PMID- 10687921 TI - Re: Flood et al. Randomized prospective study of the influence of steroids on postoperative eye-opening after exploration. PMID- 10687922 TI - Re: Flood et al. Randomized prospective study of the influence of steroids on postoperative eye-opening after exploration of the orbital floor. PMID- 10687923 TI - Double bond content of phospholipids and lipid peroxidation negatively correlate with maximum longevity in the heart of mammals. AB - Free radical damage is currently considered a main determinant of the rate of aging. Unsaturated fatty acids are the tissue macromolecules most sensitive to oxidative damage. Therefore, the presence of relatively low degrees of fatty acid unsaturation is expected in the tissues of longevous animals. In agreement with this prediction, fatty acid analyses of heart phospholipids in eight mammals ranging in maximum life span (MLSP) from 3.5 to 46 years showed that their total number of double bonds is negatively correlated with MLSP (r = -0.78, P < 0.02). The low double content of longevous mammals was not due to a low polyunsaturated fatty acid content. Instead, it was mainly due to a redistribution between types of polyunsaturated fatty acids from the highly unsaturated docosahexaenoic acid (22:6n-3) to the less unsaturated linoleic acid (18:2n-6) in longevous animals (r = -0.89, P < 0.003 for 22:6n-3 and r = 0.91, P < 0.002 for 18:2n-6 versus MLSP), where n = number of different animals in each species. This redistribution suggests that one of the mechanisms responsible for the low number of fatty acid double bonds is the presence of low desaturase activities in longevous animals, although other causing factors must be involved. In agreement with the low degree of fatty acid unsaturation of longevous mammals, the sensitivity to lipid peroxidation (r = -0.87; P < 0.005) and the in vivo lipid peroxidation (r = 0.86, P < 0.005) in the heart were also negatively correlated with MLSP across species. These results, together with previous ones obtained in rodents, birds, and humans, suggest that the low degree of tissue fatty acid unsaturation of longevous homeothermic animals could have been selected during evolution to protect the tissues against oxidative damage. PMID- 10687924 TI - Short-term calorie restriction improves disease-related markers in older male rhesus monkeys (Macaca mulatta). AB - Calorie restriction (CR) is widely known for its effects on life span, physiological aging and age-related disease in laboratory rats and mice. Emerging data from CR studies in rhesus monkeys suggest that this nutritional intervention paradigm may also have beneficial effects in long-lived mammals. Studies from our laboratory and others have suggested that young- or adult-onset CR might have beneficial effects on cardiovascular disease and diabetes. For example, long-term CR reduced body fat and serum triglycerides, and increased a subfraction of HDL cholesterol associated with decreased cardiovascular disease risk. These studies suggested that long-term CR begun in young or adult animals might have important effects on markers relevant to age-related disease. Few studies have examined the effects of CR initiated in older animals (rodents or monkeys), and the temporal nature of some potentially beneficial effects of CR is unknown. The present study examined several markers related to diabetes and cardiovascular disease in thirteen older adult (> 18 year) non-obese (body fat < 22%), male rhesus monkeys during a short-term CR paradigm. Specifically, we collected these data at baseline (ad libitum feeding), 10, 20, and 30% CR, and at 6 and 12 months on 30% CR. Fasting and peak insulin were significantly reduced as were the acute and second-phase insulin responses. CR also marginally reduced triglycerides (50% reduction), but had no effect on total serum cholesterol or blood pressure. Interestingly, the observed glucoregulatory changes emerged prior to any evidence of a change in body composition suggesting that certain effects of CR may not be wholly dependent on changes in body composition in older monkeys. PMID- 10687925 TI - Genetic differences in the age-associated decrease in inducibility of natural killer cells by interferon-alpha/beta. AB - Natural killer (NK) cells, which are important in viral infections and anti-tumor activity, show reduced cytotoxicity in aged mice. The mechanism(s) for this age related decline in NK activity has not been clearly established. We assessed changes in NK cytotoxicity in splenocytes and peripheral blood mononuclear cells after interferon (IFN)-alpha/beta stimulation in adult (6 months) and aged (22-26 months) C57Bl/6, Balb/c, and (Balb/c x C57Bl/6)F1 mice. Aged C57Bl/6 and Balb/c mice had a significantly reduced IFN-alpha/beta-stimulated NK cytotoxicity compared to adult mice. In contrast, adult and aged F1 mice showed similar NK cytotoxicity after IFN-alpha/beta induction. The decreased ability of NK cells of aged mice to respond to induction by IFN-alpha/beta was not due to a requirement for an increased amount of IFN or for a longer period of treatment with IFN. Further, this decreased response did not appear to be the result of suppressive activity of adherent cells or T cells. While the percentage of NK cells (NK1.1+) was similar in adult and aged mice, the (CD8+ NK1.1+) subset of NK cells was significantly increased in aged mice. Importantly, the percentage of CD8+ NK1.1+ cells was inversely related to the cytotoxicity observed after IFN-alpha/beta treatment. PMID- 10687927 TI - Sudden senescence syndrome plays a major role in cell culture proliferation. AB - Normal human cells of various types have a finite and predictable proliferative potential in vitro. This limited life span is due to a gradually increasing fraction of senescent cells that appear in the culture in a sudden and stochastic fashion due to a phenomenon referred to as sudden senescence syndrome (SSS). Because nondividing cells increasingly accumulate in the culture, dividing cells have to compensate for nondividers in order to accomplish additional population doubling (PD). Thus, individual dividing cells undergo more divisions, called cell generations (CG), than the number of PDs. Based on integrated experimental data, we calculated maximum CG for normal human diploid fibroblasts (HDF). It appears that for a HDF culture that undergoes 65 PD, the calculated final CG is at least 126. Based on the obtained value for CG we calculated the total size of the culture, both with and without effect of SSS. If no SSS takes place and cells divide by geometrical progression, the culture will grow up to 2(126) or 10(38) cells. By constantly eliminating cells from further divisions, causing cell loss (CL), SSS reduces the total size of the culture at every point during its proliferation. The calculated value for CL is enormous, so that the culture of 10(38) cells is reduced to only 10(19) cells, thus as little as 10(-17)% of its size! Accordingly, by preventing virtually every cell in the culture from reaching its original maximum doubling capacity, SSS appears to be the most important mechanism that influences cell culture proliferation. PMID- 10687926 TI - Mechanical stress enhances expression and production of plasminogen activator in aging human periodontal ligament cells. AB - Plasminogen activator (PA) converts plasminogen to plasmin, and plasmin activates the kinin cascade and latent extracellular matrix metalloproteases. The periodontal ligament serves to anchor the tooth to the alveolus and functions as a cushion between these hard tissues to migrate occlusal force during mastication. We reported previously that repeated mechanical tension force (MTF) as an experimental model of a traumatic occlusion, increased PA activity in human periodontal ligament derived fibroblast (hPLF) cells. In this study, the influence of in vitro cellular aging on MTF-stimulated PA activity in hPLF cells was studied. Aged hPLF cells produced a significantly higher PA activity when compared with those of young hPLF cells in response to MTF in a time- and magnitude-dependent manner. tPA mRNA levels in aged cells were higher than those in young cells, whereas PAI-1 mRNA remained unchanged and uPA mRNA was not detected. Because MTF-stimulated PA activity from hPLF cells was increased by in vitro cellular aging, aging of the periodontal ligament may affect the severity of the inflammation and the degradation of the extracellular matrix of periodontal ligament tissue by producing a large amount of PA in response to excessive force such as a traumatic occlusion. PMID- 10687928 TI - Tone burst-evoked myogenic potentials in human neck flexor and extensor. AB - Vestibular evoked myogenic potential (VEMP) has been proposed to be a manifestation of sacculocollic reflex. In a recent study using intracellular recording from neck flexor and extensor motoneurons, the neuronal connections and pathways underlying sacculocollic reflexes were determined in cats. The results showed that sacculocollic reflex displayed inhibitory connection to bilateral neck flexors and excitatory connection to bilateral neck extensors. A total of 16 normal young adults were tested with bilateral recordings of sternocleidomastoid (SCM) and splenius capitis (SC) muscles by acoustic stimulus of 500 Hz short tone burst. The results revealed that polarity of the wave I/II of VEMP on SC was the reverse of that on SCM. This implied that VEMP from ipsilateral SCM showed inhibitory neural activity; whereas VEMP from ipsilateral SC was an excitatory response. Using this non-invasive technique, the sacculocollic reflexes in human neck flexor and extensor were studied. The results in humans were consistent with the previous findings in cats. PMID- 10687929 TI - Benign paroxysmal positional vertigo (BPPV): idiopathic versus post-traumatic. AB - Between October 1974 and August 1997 in our Dizziness Clinic (n = 15,233), 2,523 patients were found to suffer from BPPV. All patients were assessed and followed up by the author. Patients (n = 337) having other ear or neurological diseases were excluded from this retrospective study. In 1644/2186 (75.21%) patients, the type of nystagmus was clinically identified in two opposite directions of gaze in the provocative head position. These patients were divided into two groups: i) idiopathic (n = 1,490) (no apparent cause); ii) post-traumatic (n = 154) (time of onset related to accident). It was found that in the idiopathic group men were older than women; women were more affected than men (2.3:1), and in the post traumatic group there was no age difference between men and women; women and men were equally affected (1:1). In addition: i) patients were older in the idiopathic than the post-traumatic group. ii) BPPV of the posterior (PSC) was by far more prevalent than BPPV of the horizontal semicircular canal (HSC) in both groups, although there was no difference in prevalence between the two groups. iii) Bilateral involvement was more prevalent in the post-traumatic group. iv) All bilateral cases in both groups suffered from BPPV of the PSC. It is concluded that despite similarities, these two groups differ in a number of parameters. Thus the pathophysiology and the course of idiopathic vs post-traumatic BPPV may also be different. PMID- 10687930 TI - Vestibulo-ocular responses during static head roll and three-dimensional head impulses after vestibular neuritis. AB - This study aimed to investigate whether unilateral vestibular neuritis (VN) causes the same deficits of ocular counter-roll during static head roll (OCR(S)) and dynamic vestibulo-ocular reflex gains during head impulses (VOR(HI)) as unilateral vestibular deafferentation (VD). Ten patients with acute and 14 patients with chronic vestibular paralysis after VN were examined. The testing battery included fundus photography of both eyes with the head upright (binocular cyclorotation) and dual search coil recordings in a three-field magnetic frame. With one dual search coil on the right eye and the other on the forehead, the following stimuli were given: i) Halmagyi-Curthoys head impulses about the vertical, horizontal and torsional axes. ii) Static roll positions of the head up to 20 degrees right- and left-ear-down by movement of the neck. The comparison group consisted of 19 healthy subjects. Compared with the VD-patients, as reported in the literature, acute VN-patients showed the same pattern of OCR(S) gain reduction and binocular cyclorotation (CRb). The main feature that distinguished chronic VN-patients from chronic VD-patients was the normalization of the torsional VOR(HI) gain to the affected side, whereas the VOR(HI) gains in the horizontal and vertical directions did not show recovery (as in the patients with chronic VD). Chronic VN-patients differed from acute VN-patients by: i) symmetrical OCR(S) gains, ii) a less pronounced CRb toward the affected side, and iii) a normal torsional VOR(HI) gain toward the affected side. Since the ipsilesional torsional VOR(HI) gain did not recover in VD-patients, the normalization of this gain in our VN-patients can only be explained by a (partial) recovery of otolith function on the side of the lesion after the neuritis. PMID- 10687931 TI - Hypotension and sensorineural hearing loss: a possible correlation. AB - A possible role of hypotension in the genesis of sudden or slowly developing sensorineural hearing loss has been outlined. In order to confirm this hypothesis, and to exclude other vascular risk factors, a prospective study was carried out within the "Brisighella Study", a wide and homogeneous group of subjects thoroughly examined from a metabolic and cardiovascular point of view. Among them, 20 participants aged 50 years or less (18 women, 2 men) with diastolic blood pressure < or = 60 mmHg and/or systolic blood pressure < or = 105 mmHg were selected and underwent otological and audiometric examinations. Patients with previous audiological, vestibular and otological diseases were excluded. The control group was represented by 100 subjects (60 women, 40 men), aged 50 years or less, randomly chosen from within a sample of the normal population in the same region. A statistically significant incidence of sensorineural hearing loss was recorded in the study group (7/20 subjects, all affected by low-frequency hearing loss), while hearing impairment was observed in only 3/100 participants in the control group. The mean values of the main metabolic parameters were normal. An alteration of the vasomotor system associated with a hypotensive condition could be responsible as a possible factor in the origin of a cochlear damage and the consequent sensorineural hearing loss. PMID- 10687932 TI - Clinical utility of LDL-apheresis in the treatment of sudden hearing loss: a prospective, randomized study. AB - Although the pathogenesis of sudden hearing loss (SHL) is not as yet known, the clinical picture and the frequent association with vascular risk factors make an ischaemic event likely. This study aimed to assess the effect of an extracorporeal procedure (H.E.L.P.) in removing LDL-cholesterol, fibrinogen and lipoprotein (a) from the plasma, on the recovery of hearing SHL. This procedure using the HELP-system was compared with the usual standard treatment with prednisolone, dextranes and pentoxifyllin. We undertook a single centre, prospective, randomized study in which 18 patients were assigned to H.E.L.P. apheresis and 9 patients were assigned to standard treatment (2:1 randomization). Audiometric and laboratory testing was performed at baseline, 24 h and 6 weeks after start of treatment. Primary endpoint was the improvement of the average pure-tone threshold between 0.125 and 8 kHz after 24 h. Twenty-four hours after H.E.L.P. treatment average pure-tone threshold recovered by 10.4 dB and by 26.4 dB after 6 weeks. The recovery of hearing of the standard treated patients was 5.8 dB and 16.8 dB after 24 h and 6 weeks respectively. LDL-cholesterol, fibrinogen and lipoprotein (a) were significantly reduced in the HELP treated patients compared with standard therapy, resulting in a significant improve in plasma viscosity, erythrocyte aggregation and resistance to oxidative stress of LDL particles. Our results suggest that the clinical outcome of SHL after a single extracorporeal LDL-apheresis is superior or at least equal to the more expensive standard treatment with prednisolone, dextranes and pentoxifyllin. Re establishment of vascular endothelial function and improved blood rheology may be the underlying cause. These results must be confirmed in larger-scale trials. PMID- 10687933 TI - Evaluation of inner ear histology and auditory brainstem response in Wriggle Mouse Sagami. AB - Wriggle Mouse Sagami (WMS) is a spontaneous mutant strain with neuroepithelial defects. These animals are characterized by abnormal movements linked to an autosomal recessive gene. To determine the association between inner ear histology and hearing ability, we assayed these characteristics in mice homozygous and heterozygous for the mutation, as well as in wild-type animals. In homozygotes, the cochlea and saccule degenerated 3 months after birth. Beginning at 3 months of age, and progressing in an age-dependent manner, the organ of Corti disappeared and the number of spiral ganglion cells decreased, starting at the basal turn and moving toward the apical turn. The sensory epithelium became atrophic in the saccule. Three-month-old heterozygotes demonstrated degeneration in the cochlea, not in the saccule. No obvious auditory brainstem evoked response (ABR) was observed at any frequency in homozygotes aged 1 month and older. In contrast, the heterozygotes retained some hearing acuity until the age of 1 month, after which they became deaf. These findings suggest that WMS mice may provide a good model that will be useful in identifying deafness genes in humans. PMID- 10687934 TI - Cochlear hook anatomy: evaluation of the spatial relationship of the basal cochlear duct to middle ear landmarks. AB - The cochlear hook is an important anatomical area for the otologist performing cochlear implants and other otological procedures, who requires knowledge of the basal cochlea. A total of 15 human temporal bones were dissected and the spatial relationship of the hook segment of the cochlear duct to the stapes, round window, cochleariform process and ductus reuniens were evaluated. Inter individual variability was noted for widths of scala tympani (average width 1.36 +/- 0.25 mm) and scala vestibuli (average width 1.18 +/- 0.18 mm) in the region of typical cochlear implant placement, with the scala vestibuli occasionally being wider than the scala tympani. The cochlear duct was in closest proximity to the stapes at the midportion of the footplate, with an average distance of 1.23 mm at this narrowest width. A fibrous anchor, not previously described in otology literature, was identified securing the most basal end of the cochlear duct. Knowing the spatial relationship of the cochlear duct to the middle and inner ear structures could prevent damage to the basilar membrane in procedures around or involving the basal cochlear, such as cochlear implantation, stapedotomy, or implantable hearing devices. PMID- 10687935 TI - Morphological changes induced by administration of a Na+,K+-ATPase inhibitor in normal and hydropic inner ears of the guinea pig. AB - The objective of this study was to determine the effects of ouabain, a Na+,K+ ATPase inhibitor, in inner ears. Administering ouabain locally through the round window and vestibule, resulted in degenerative changes in cochlear and vestibular sensory cells and limbal fibrocytes, but the stria vascularis and spiral ligament were less affected. The position of Reissner's membrane was rarely changed. Vacuolar spaces in the sensory epithelia of cristae, maccula utriculi and macula sacculi increased in number. Nystagmus was a common occurrence with or without demonstrating degeneration of vestibular sensory cells. By administering ouabain systemically, the course of developing endolymphatic hydrops could not be altered in the ears with endolymphatic duct blockage. Edema of nerve endings of inner hair cells and vestibular sensory cells was frequently observed with administration of a high concentration of ouabain in both normal and hydropic ears, but edema was reversible. Degeneration of some vestibular sensory cells were observed in hydropic ears with a long survival time. The mechanism of selective sensitivity or non-sensitivity of inner ear tissues to ouabain is discussed. PMID- 10687936 TI - Improved RNA analysis for immediate autopsy of temporal bone soft tissues. AB - RNA analysis is essential for understanding biological activities of a cell or tissue. Unfortunately, retrieval of RNA from existing archives of human temporal bones has proven extremely difficult due to degradation of RNA molecules. The major factors that contribute to degradation of RNA in specimens from autopsied temporal bones are tissue autolysis due to time elapsed before autopsy, and technical problems in processing the bones after harvest. We therefore focused on improving the survival of RNA in human temporal bones by shortening the time to autopsy and through modification of the processing technique by removing targeted tissues directly from the temporal bones and by avoiding time-consuming decalcification and celloidin-embedding. Eight temporal bones collected at immediate autopsies were used in this study. Representative mRNAs, ranging from high (MUC5B, physically unstable) to low (beta-actin, physically stable) molecular weights, and from abundant (MUC5B) to non-abundant (MUC1) RNA, were studied by in situ hybridization, Northern blot technique, or both. Using this modified protocol in autopsies performed up to 6 h after death, the existence of mRNAs was demonstrated in all bones studied. This improved method demonstrates the feasibility of the use of autopsied temporal bone tissues for RNA analysis. PMID- 10687937 TI - Management of intrameatal vestibular schwannoma. AB - The growth of purely intrameatal vestibular schwannoma (VS) was investigated, in the period 1973-96 in a series of 40 patients with 40 unilateral VS. In the present study, the material was analysed and updated. By the end of the observation period (mean 3.6 years), 27 tumours (67.5%) revealed growth and 13 tumours (32%) had no measurable growth. Four growth patterns were observed: (A) 15 tumours (37.5%) exhibited constant growth; (B) 13 tumours (32.5%) had no measurable growth; (C) 8 tumours (20%) revealed growth subsequent to a no-growth period; and (D) 4 tumours (10%) showed different growth patterns during the observation period. The annual diameter growth rate ranged between 00 mm/year and 6.5 mm/year and the mean diameter growth per year was 3.2 mm. The findings of the present study, especially those for group B (the non-growing tumours) and C (tumour growth subsequent to a silent period) bring into question the reliability of the results achieved by radiosurgery, as without any intervention it may be that no tumour growth occurs. PMID- 10687938 TI - Expression of vascular endothelial growth factor in otitis media. AB - Increased vascular permeability and endothelial cell growth are important in the pathogenesis of otitis media with effusion (OME) and the vascular endothelial growth factor (VEGF) is known to play an important role in the increased vascular permeability and angiogenesis. To date, at least five isoforms of the VEGF family have been identified as VEGF transcripts, encoding polypeptides of 206, 189, 165, 145 and 121, but their physiological roles are unclear. The purpose of this study was to investigate the expression of VEGF, in both endotoxin-induced OME of the rat and human otitis media. We instilled endotoxin and saline as a control into the middle ear cavity of the rat. Middle ear mucosa were taken at 0 h, 1 h, 3 h, 6 h, 12 h, 1 day, 3 days, 7 days and 14 days and the expression of VEGF mRNA and VEGF protein was evaluated using semi-quantitative RT-PCR and immunohistochemistry. Expression of VEGF164 mRNA and VEGF120 mRNA was first identified 1 h after endotoxin instillation and was dramatically increased over the period 6 h-1 day and then progressively decreased by day 7. The level of expression of VEGF120 mRNA was slightly higher than that of VEGF164 mRNA and that of VEGF164 mRNA was much higher than that of VEGF188 mRNA. Immunostaining revealed expression of VEGF during 6 h to day 3 and its expression was localized to ciliated cells and some inflammatory cells. We also performed RT-PCRs of cDNA from middle ear fluids of 8 human OME patients and middle ear mucosa of 4 chronic otitis media patients for the identification of VEGF mRNA expression. VEGF121 mRNA was highly expressed in all samples compared with VEGF165 mRNA. These results suggest that VEGF may be primarily responsible for increased vascular permeability and endothelial cell growth in OME and that VEGF seems to play a significant role in the pathogenesis of OME. PMID- 10687939 TI - Continuous long-term measurements of the middle ear pressure in subjects with symptoms of patulous eustachian tube. AB - Direct middle ear (ME) pressure measurements during 24 h and tubal function tests were performed on 11 subjects with the clinical diagnosis patulous Eustachian tube (PET). The pressure was recorded from the ME via a perforation in the tympanic membrane. The method used has previously been reported in a study on subjects with normal ME function and no symptoms of PET: a normal group. Results from tubal function tests correlated well with the clinical diagnosis of PET and the sniff test was positive in most subjects. Continuous ME pressure measurements during normal everyday and night-time activities showed no indications of pressure changes induced by sniffing. Subjects with the clinical diagnosis PET did not have a static state of an open ET. The ME pressure varied during the day and night, indicating that the function of the ET changed over time from a closed to an open state. Subjects with PET had a long-term pressure pattern significantly different from that of a normal group, a difference seen in the erect as well as in the recumbent position during the night. The subjects demonstrated a negative mean pressure level in the recumbent position during the night, whereas the normal group had a slight positive mean pressure. The difference was statistically significant. Many subjects demonstrated frequent slow rate negative pressure trends during normal day-time and night-time activities. Long-term continuous pressure measurements seem to add valuable information to that of conventional tubal function tests. PMID- 10687940 TI - Eotaxin synthesis by nasal polyp fibroblasts. AB - Nasal polyps is a chronic inflammatory disease of the upper airway characterized by structural abnormalities including stromal fibrosis. Fibroblasts are a rich source of cytokines and inflammatory mediators and are thought to play an important role in the development of fibrosis. In addition, there is considerable evidence for the participation of eosinophils in the pathophysiology of nasal polyps. Although increased numbers of eosinophils are present in nasal polyps, the mechanisms responsible for their selective accumulation are not completely clear. Eotaxin is a chemokine that promotes the selective recruitment of eosinophils. Thus, it may be an important molecule for the recruitment of eosinophils in nasal polyps. The purpose of this study was to investigate whether nasal polyp fibroblasts synthesize eotaxin after stimulation with lipopolysaccharide, IL-1beta or TNF-alpha. Using primary nasal polyp tissue derived fibroblast lines, we demonstrated that LPS, IL-1beta and TNF-alpha induced the gene expression and protein production of eotaxin in nasal polyp fibroblasts. This responsiveness to LPS, IL-1beta and TNF-alpha was time- and dose-dependent. These findings support the hypothesis that fibroblasts could play an important role in the recruitment of eosinophils in nasal polyps through the production of eotaxin. PMID- 10687941 TI - Ciliostimulatory effects mediated by nitric oxide. AB - Ciliostimulation induced by various transmitters has been suggested to be mediated by the release of nitric oxide (NO). Freshly obtained adenoid tissue explants were pre-treated with the nitric oxide synthase (NOS) inhibitor N(G) nitro L-arginine (L-NNA), to determine whether the ciliostimulators terbutaline, methacholine, substance P, and endothelin-1 require the release of NO to increase ciliary beat frequency (CBF) in vitro. The L-NNA pre-treatment affected the change in CBF induced by each of the ciliostimulators tested. To determine whether cyclic nucleotides also stimulate CBF by inducing the release of NO, an extra series of experiments were performed with dibutyryl cAMP and dibutyryl cGMP, and L-NNA pre-treatment. In contrast to the experiments with the various ciliostimulators, both dibutyryl cAMP and dibutyryl cGMP exerted ciliostimulatory effects that could not be inhibited by L-NNA. The present findings suggest that NO acts as an intermediate messenger in the ciliated epithelium in response to various transmitters and mediators. On the other hand, pre-treatment with the NOS inhibitor L-NNA did not affect ciliary response to the second messengers cAMP and cGMP, thus suggesting that NO dependent mechanisms do not constitute the sole pathway for the stimulation of ciliary function. PMID- 10687942 TI - Microsurgical endonasal decompression in dysthyroid orbitopathy. AB - Diagnosis of thyroid eye disease can be established by its history, signs, symptoms, clinical and laboratory findings of an autoimmune thyroid disease. Therapy for this disease is limited to a few options, which should be administered depending on its stage and inflammatory activity. When medication and radiation therapy fail indications for decompression are: loss of visual acuity or visual field defects, increasing strabism and severe keratopathy due to eyelid retraction. Numerous surgical decompression techniques have been described in endocrine orbitopathy. We have adopted endonasal microsurgery, because this technique gives the freedom to work bimanually, ensures a stereomicoscopic view of the intranasal landmarks of orbital walls and allows simultaneous decompression of the medial and inferior orbital wall as well as a good relief of pressure at the orbital apex. Decompressions were performed on 27 orbits in 17 patients, via the endonasal microsurgical, 3 via external approach. The microscopic approach was entirely comparable with regard to reduction of proptosis with a mean improvement of 4.1 mm against a mean of 4.7 mm by external approach and a mean 0.2 of better visual acuity in both procedures. The microsurgical technique is considered superior to an external approach avoiding external scars, neural pains and reportedly less diplopia. Also, trauma to the nalolacrimal and nasofrontal ducts are avoided. The healing phase and the hospitalization time is shorter. PMID- 10687943 TI - Bacterial interference in the nasopharynx and nasal cavity of sinusitis prone and non-sinusitis prone children. AB - The aim of this study was to compare the frequency of recovery of potential pathogens and aerobic and anaerobic interfering bacteria in the nasopharynx and nasal cavity of sinusitis prone (SP) children, with their recovery in non sinusitis prone (N-SP) children. Nasopharyngeal and nasal cultures were taken from 20 SP and 20 N-SP children. Potential pathogens and aerobic and anaerobic bacteria with interfering capabilities against these micro-organisms were identified. Twenty-one potential pathogens (1.05 patient) were isolated from nasopharyngeal cultures from 14 of the 20 SP children, and 10 (0.5 patient) were recovered from 6 of the 20 NSP (p < 0.05). Bacterial interference between two aerobic (alpha and non-haemolytic streptococci) and two anaerobic species (Prevotella and Peptostreptococcus species) and four potential pathogens was observed. Bacterial interference was noted in 64 instances against 4 potential pathogens by 24 normal flora isolates that were recovered from 7 of the SP group and in 144 instances by 47 isolates from 18 of the NSP group (p < 0.05). Nineteen potential pathogens (0.95/patient) were isolated from nasal cultures of 13 of the 20 SP children and 8 (0.4/patient) were recovered from 5 of the 20 NSP (p < 0.05). Bacterial interference by similar micro-organisms was noted in 21 instances by 9 normal flora isolates that were recovered from 5 of the SP group, and in 63 instances by 26 isolates from 15 of the NSP group (p < 0.05). Our findings illustrate for the first time that the nasopharyngeal and nasal flora of NSP children contains more aerobic and anaerobic micro-organisms with interfering capability and less potential pathogens than that of SP children. PMID- 10687944 TI - Effects of sustained-release oral phenylpropanolamine on the nasal mucosa of healthy subjects. AB - Phenylpropanolamine (PPA) is widely used as a nasal decongestant administered orally in sustained release preparations and, in Sweden, the recommended dose nowadays is 50 mg twice daily for adults. The aim of this placebo-controlled, cross-over study was to determine the onset and duration of the decongestive effect of 50 and 100 mg PPA in 15 healthy subjects. All subjects arrived at the laboratory at 07.30 h. After an acclimatisation, the nasal mucosal baseline was established with rhinostereometry and the minimal cross-sectional area was measured using acoustic rhinometry. The systolic and diastolic blood pressures were also determined. Then all subjects were given their study drugs for the day and the measurements were repeated every hour for 8 h. This procedure was repeated for 3 days at 48 h intervals between the days. For purposes of comparison, the decongestive effect of oxymetazoline nasal spray was studied on a separate day. The decongestive effect of 100 mg PPA was similar to that of topical oxymetazoline. It develops after 1 h and lasts for approximately 6 h. The decongestive effect of oxymetazoline was significantly greater than that of 50 mg PPA and that of 100 mg PPA was significantly greater than that of 50 mg PPA using rhinostereometry, but not when using acoustic rhinometry. However, 50 mg PPA had no significant decongestive effect, compared with placebo, with rhinostereometry or acoustic rhinometry. In the first 3 h after administration of PPA, there was a dose-response increase in the systolic and diastolic blood pressures, which then returned to baseline. In conclusion, this study shows that PPA in double the recommended dose, i.e. 100 mg, has a significant decongestive effect on the nasal mucosa in healthy subjects. However, when the dose of PPA is increased the systolic and diastolic blood pressures also increase. PMID- 10687945 TI - Postnatal changes in the types of muscle fibre in the canine inferior pharyngeal constrictor. AB - Deglutition is considered to be immature in infants and to mature postnatally. We evaluated postnatal changes in muscle fibre type composition in the canine inferior pharyngeal constrictor muscle, which consists of the thyropharyngeal (TP) and cricopharyngeal (CP) muscles, using ATPase staining with respect to the maturation of deglutition. In the TP muscle type IIA and type IIB fibres, the main components in the adult, were already predominant at 1 week postnatally. The percentage of primitive type IIC fibre showed a rapid reduction and reached the adult level within 6 weeks. In the CP muscle, the majority of fibres were type IIC at 2 weeks. At 2 months, more than 20% of the fibres were still type IIC and the proportion of type I fibres as a main component in the adult was smaller than that of the adult. None of the puppies younger than 9 weeks old had a fibre type composition similar to that of the adult. In the extensor digitorum longus and flexor digitorum superficialis, the compositions of muscle fibre types became similar to that of the adult at 6 and 9 weeks of age, respectively. Thus, the TP muscle matured more rapidly than the limb muscles, while the CP muscle matured more slowly. We speculated that the TP and CP muscles have specific individual differentiation patterns associated with their functional roles before and after birth, compared with the limb muscles. PMID- 10687946 TI - Glottic and supraglottic laryngeal carcinoma: differences in epidemiology, clinical characteristics and prognosis. AB - In order to evaluate differences in epidemiology, clinical characteristics and prognosis, 166 glottic and 127 supraglottic cases of laryngeal squamous cell carcinoma diagnosed between 1962 and 1991 at Tampere University Hospital, Finland, were reviewed. The annual age-adjusted incidence in males decreased from 6.7/100,000 to 2.6/100,000 and the proportion of glottic tumours increased from one-third to two-thirds during the study period. The proportion of early stage lesions was greater among glottic tumours, and patients with a supraglottic tumour presented more often with neck node metastases. Hoarseness was the most common symptom, being more prevalent in patients with a glottic tumour. The symptom pattern of supraglottic carcinoma was altogether more diffuse. The 5-year disease-specific survival was 81% in glottic and 70% in supraglottic disease, but the difference in survival was not statistically significant. In the multivariate Cox regression analysis, higher T-category and presence of neck node metastases had adverse prognostic effect, while location of the tumour did not significantly affect the prognosis. Favourable changes in smoking habits seem to be the main reason for the incidence decrease and obviously also for the decrease in the proportion of supraglottic tumours. PMID- 10687947 TI - Fibroblast growth factor-2. AB - Fibroblast growth factor-2 (FGF-2) is a member of a large family of proteins that bind heparin and heparan sulfate and modulate the function of a wide range of cell types. FGF-2 stimulates the growth and development of new blood vessels (angiogenesis) that contribute to the pathogenesis of several diseases (i.e. cancer, atherosclerosis), normal wound healing and tissue development. FGF-2 contains a number of basic residues (pI 9.6) and consists of 12 anti-parallel beta-sheets organized into a trigonal pyrimidal structure. FGF-2 binds to four cell surface receptors expressed as a number of splice variants. Many of the biological activities of FGF-2 have been found to depend on its receptor's intrinsic tyrosine kinase activity and second messengers such as the mitogen activated protein kinases. However, considerable evidence suggest that intracellular FGF-2 might have a direct biological role particularly within the nucleus. In addition, heparan sulfate proteoglycans have been demonstrated to enhance and inhibit FGF-2 activity. The possibility that FGF-2 activity can be manipulated through alterations in heparan sulfate-binding is currently being exploited in the development of clinical applications aimed at modulating either endogenous or administered FGF-2 activity. PMID- 10687948 TI - Caspase-9. AB - Caspase-9 is a member of caspase family of cysteine proteases that have been implicated in apoptosis and cytokine processing. When cells receive apoptotic stimuli, mitochondria releases cytochrome c which then binds to Apaf-1, the mammalian Ced-4 homologue, together with dATP. The resultant complex recruits Caspase-9 leading to its activation. Activated Caspase-9 cleaves downstream caspases such as Caspase-3, -6 and -7 initiating the caspase cascade. The majority of homozygous Caspase-9 null mice die perinatally with a markedly enlarged and malformed cerebrum caused by a reduction of apoptosis during early brain development. Thus, Caspase-9 function is essential for apoptosis during normal development of the central nervous system. These data suggest that inhibition of Caspase-9 activity would render opportunity to treat patients suffering from neurological diseases such as stroke, neurodegenerative diseases or brain injury caused by hypoxia. PMID- 10687949 TI - Bikunin--not just a plasma proteinase inhibitor. AB - Bikunin is a plasma proteinase inhibitor that has received little attention in the past, probably because its activity towards various proteinases was found to be relatively weak in early work. It was recently discovered, however, that bikunin effectively inhibits a proteinase that seems to be involved in the metastasis of tumour cells--cell surface plasmin--and that a fragment of bikunin inhibits two proteinases of the coagulation pathway--factor Xa and kallikrein. Furthermore, it has been found that bikunin has other properties, such as the ability to modulate cell growth and to block cellular calcium uptake. Most of the bikunin in the blood occurs as a covalently linked subunit of the proteins pre- and inter-alpha-inhibitor. In this form bikunin lacks some of its known activities, and there is evidence that its release by partial proteolytic degradation may function as a regulatory mechanism. Although the physiological function of bikunin still remains to be established, current data suggest that this protein plays a role in inflammation. Further studies could therefore lead to results of therapeutical value. PMID- 10687950 TI - The quest to deduce protein function from sequence: the role of pattern databases. AB - In the wake of the numerous now-fruitful genome projects, we have witnessed a 'tsunami' of sequence data and with it the birth of the field of bioinformatics. Bioinformatics involves the application of information technology to the management and analysis of biological data. For many of us, this means that databases and their search tools have become an essential part of the research environment. However, the rate of sequence generation and the haphazard proliferation of databases have made it difficult to keep pace with developments, even for the cognoscenti. Moreover, increasing amounts of sequence information do not necessarily equate with an increase in knowledge, and in the panic to automate the route from raw data to biological insight, we may be generating and propagating innumerable errors in our precious databases. In the genome era upon us, researchers want rapid, easy-to-use, reliable tools for functional characterisation of newly determined sequences. For the pharmaceutical industry in particular, the Pandora's box of bioinformatics harbours an information-rich nugget, ripe with potential drug targets and possible new avenues for the development of therapeutic agents. This review outlines the current status of the major pattern databases now used routinely in the analysis of protein sequences. The review is divided into three main sections. In the first, commonly used terms are defined and the methods behind the databases are briefly described; in the second, the structure and content of the principal pattern databases are discussed; and in the final part, several alignment databases, which are frequently confused with pattern databases, are mentioned. For the new-comer, the array of resources, the range of methods behind them and the different tools required to search them can be confusing. The review therefore also briefly mentions a current international endeavour to integrate the diverse databases, which effort should facilitate sequence analysis in the future. This is particularly important for target-discovery programmes, where the challenge is to rationalise the enormous numbers of potential targets generated by sequence database searches. This problem may be addressed, at least in part, by reducing search outputs to the more focused and manageable subsets suggested by searches of integrated groups of family-specific pattern databases. PMID- 10687951 TI - Role of reactive oxygen species in apoptosis: implications for cancer therapy. AB - Reactive oxygen species are widely generated in biological systems. Consequently humans have evolved antioxidant defence systems that limit their production. Intracellular production of active oxygen species such as *OH, O2- and H2O2 is associated with the arrest of cell proliferation. Similarly, generation of oxidative stress in response to various external stimuli has been implicated in the activation of transcription factors and to the triggering of apoptosis. Here we review how free radicals induce DNA sequence changes in the form of mutations. deletions, gene amplification and rearrangements. These alterations may result in the initiation of apoptosis signalling leading to cell death, or to the activation of several proto-oncogenes and or the inactivation of some tumour suppressor genes. The regulation of gene expression by means of oxidants, antioxidants and the redox state remains as a promising therapeutic approach. Several anticarcinogenic agents have been shown to inhibit reactive oxygen species production and oxidative DNA damage, inhibiting tumour promotion. In addition, recombinant vectors expressing radical-scavenging enzymes reduce apoptosis. In conclusion, oxidative stress has been implicated in both apoptosis and the pathogenesis of cancer providing contrived support for two notions: free radical reactions may be increased in malignant cells and oxidant scavenging systems may be useful in cancer therapy. PMID- 10687952 TI - Bi-directional signal transduction by integrin receptors. AB - The integrin family of cell surface glycoproteins functions primarily as receptors for extracellular matrix ligands. There are now many well characterized integrin-ligand interactions which are known to influence many aspects of cell behaviour including cell morphology, cell adhesion, cell migration as well as cellular proliferation and differentiation. However, in fulfilling these functions, integrins are not simple adhesion receptors that physically mediate connections across the plasma membrane. Rather, integrin function itself is highly regulated, largely through the formation of specific associations with both structural and regulatory components within cells. It is these intracellular interactions which allow integrin function to effect many biochemical signalling pathways and therefore to impinge upon complex cellular activities. Recently, much research has focused on elucidating the molecular mechanisms which control integrin function and the molecular processes which transduce integrin-mediated signalling events. In this review, we discuss progress in the field of integrin signal transduction including, where applicable, potential therapeutic applications arising from the research. PMID- 10687953 TI - Binding of 125I-insulin-like growth factor-II to cells cultured in fetal bovine serum: a complication. AB - Insulin-like growth factor II is an important fetal mitogen in mice and humans and its biological activity is regulated in a complex manner. The peptide interacts with three membrane-bound receptors, with a superfamily of insulin-like growth factor binding proteins and with the proteoglycan, glypican-3. Recently, the blood protein, vitronectin, has been identified as a novel insulin-like growth factor II-binding protein. Many studies have used cell lines maintained in fetal bovine serum to identify cell surface insulin-like growth factor II binding sites. We now describe a complication associated with the interpretation of such in vitro studies. Fetal bovine serum-derived vitronectin adheres very tightly to tissue culture dishes. When cells that have been maintained in fetal bovine serum are incubated with 125I-insulin-like growth factor II, a substantial fraction of the 125I-insulin-like growth factor II apparently associated with the cell surfaces may represent radioliogand bound by the fetal bovine serum-derived vitronectin. This may result in over-estimation of cell surface insulin-like growth factor II binding sites. PMID- 10687954 TI - Characterisation of fibrillin-1 cDNA clones in a human fibroblast cell line that assembles microfibrils. AB - Fibrillin-1 is a large extracellular glycoprotein which is a major structural component of 10-12 nm microfibrils. Defects in human fibrillin-1 give rise to the autosomal dominant connective tissue disease the Marfan syndrome and related disorders. Previous studies examining the biosynthesis and secretion of recombinant fibrillin-1 fragments have been performed in cell lines which do not assemble fibrillin into extracellular 10-12 nm microfibrils. Conflicting data have been obtained regarding N-terminal processing. In this study we have characterised a human fibroblast cell line MSU-1.1 which shows a similar endogenous fibrillin-1 pulse chase profile to primary human dermal fibroblasts and produces microfibrils. Expression of a approximately 50 kDa N-terminal recombinant peptide in MSU-1.1 resulted in efficient secretion of this peptide into conditioned media, N-terminal sequence analysis of the purified peptide identified 2 protease cleavage sites and a presumed signal peptidase site. Together these data identify the natural leader sequence of fibrillin-1 and the presence of two processing sites in the N-terminus of fibrillin-1. The identification of an N-terminal processing site in recombinant fibrillin-1 similar to that obtained in a previous study which used an HT1080 fibrosarcoma host cell line excludes defective N-terminal processing as the cause of the assembly defect in this cell line. A full length normal and mutant fibrillin cDNA (approximately 8.6 kb) was constructed and stable integration of each into MSU1.1 led to RNA transcription at approximately 5% of endogenous levels. This is the first report of transcription from the full length fibrillin-1 cDNA. The low levels of transcription achieved, suggest that additional upstream and downstream DNA sequence elements will be required for high levels of full length fibrillin-1 cDNA expression. PMID- 10687955 TI - The overexpression of the CDC25 gene of Saccharomyces cerevisiae causes a derepression of GAL system and an increase of GAL4 transcription. AB - The CDC25 gene product is an exchange factor for Ras proteins and it activates the Ras/cAMP pathway in the yeast Saccharomyces cerevisiae. The overexpression of the CDC25 gene in S. cerevisiae cells causes a partial glucose-derepressed phenotype which is particularly evident for expression of invertase. To define domains of Cdc25 protein relevant for this derepression and to test another glucose repressed system, different to invertase, we have overexpressed different regions of the CDC25 gene under the control of a GAL-promoter. We found that a derepression of both GAL regulated promoters and invertase was related to the overexpression of CDC25 regions that contain a functional guanine nucleotide exchange (GEF) domain. The effect on GAL-promoters was particular evident when the CDC25 gene was under the control of a UASgal element and operates at transcriptional level, although a moderate derepression was found also for UASgal/lacZ reporter gene. Finally, the overexpression of the GEF domain of CDC25 also caused an increase in the expression of the GAL4 regulatory gene, while a constitutive activation of the Ras/cAMP pathway did not produce any increase in GAL4 expression. These findings indicate that the overexpression of the catalytic domain of CDC25 gene is necessary and sufficient to give a glucose-derepression of GAL promoters and of invertase. They also suggest that the derepression of GAL promoters occurs through an increase of GAL4 expression in a Ras cAMP independent way. PMID- 10687956 TI - The plasma membrane of Xenopus laevis oocytes contains voltage-dependent anion selective porin channels. AB - Recent patch-clamp studies have shown that anti-porin antibodies, applied to the external side of excised plasma membrane patches of mammalian astrocytes, close chloride channels that are thought to be engaged in cell volume regulation. Frog oocytes are often used to study this basic cell function. Here we document the localisation of endogenous porin voltage-dependent anion-selective channels in Xenopus laevis oocyte plasma membranes. In confocal laser microscopy images a disjunctive pattern of fluorescing spots appear about 10 microm apart. Labelling was prevented by preabsorption of the antibodies with synthetic peptides comprising the epitope of the antigen. Immuno-gold marking of oocyte surfaces followed by silver enhancement of the gold particles lead to a plasma membrane labelling corresponding to that obtained by the confocal laser approach. The data suggests the presence of voltage-dependent, anion-selective channels in oocyte plasma membranes. This data should be borne in mind when frog oocytes are used to study the characteristics of endogenous or heterologously expressed ion channels or regulatory proteins. PMID- 10687957 TI - Ribonuclease, cell-free translation-inhibitory and superoxide radical scavenging activities of the iron-binding protein lactoferrin from bovine milk. AB - The purpose of this study was to characterize the ribonuclease (RNase) and cell free translation-inhibitory activities of lactoferrin isolated from bovine milk. It was found that bovine lactoferrin exhibited ribonucleolytic activity toward yeast transfer RNA in a dose-dependent manner. The pH optimum for this RNase activity was in the vicinity of 7.5. Lactoferrin exerted RNase activity on poly C with an activity of 2.15 U/mg. No activity was detected toward poly A, poly G, and poly U. The milk protein inhibited cell-free translation in rabbit reticulocyte lysate with an IC50 of 9.6 microM. The protein was devoid of N glycosidase activity characteristic of ribosome inactivating proteins which also possess RNase and cell-free translation-inhibitory activities. It inhibited superoxide radical formation. PMID- 10687958 TI - Detection of a mammalian histone H4 kinase that has yeast histidine kinase-like enzymic activity. AB - A well characterized histidine kinase purified from yeast has been shown to phosphorylate histone H4 on a histidine residue. This enzyme is unlike the two component histidine kinases predominantly found in prokaryotes. Until now, a histidine kinase similar to this yeast enzyme has not been purified from a mammalian source. By using a purification scheme similar to that used to purify the yeast histidine kinase, a protein fraction with histone H4 kinase activity has been isolated from porcine thymus. The yeast histidine kinase was shown to be detectable using an in-gel kinase assay system and using this system, four major bands of histone H4 kinase activity were apparent in the porcine thymus preparation. Through the use of immunoprecipitation, alkaline hydrolysis and subsequent phosphoamino acid analysis it has been demonstrated that this partially purified kinase fraction is capable of phosphorylating histone H4 on histidine. In conclusion, an preparation has been made from porcine thymus that contains histone H4 kinase activity and at least one of the kinases present in this preparation is a histidine kinase. PMID- 10687959 TI - Presence of uterine peroxidase activity in the rat early pregnancy. AB - Peroxidase has been associated with estrogen action in the uterus. This enzyme plays an important role in the control of hydrogen peroxide levels and in catechol estrogen production. Since the uterus, during early pregnancy, is subjected to estrogen and progesterone regulation, we analyzed the changes of peroxidase activity in relation to receptivity and uterine early response to the embryo. Soluble and microsomal peroxidase activity were determined in the rat uterus during the estrus phase and early pregnancy (days 3 through 6). Soluble peroxidase activity increased significantly (p < 0.01) from day 3 (1.50 +/- 0.24) to day 4 (3.5 +/- 0.3) and 5 (5 +/- 0.5 U/mg protein, mean +/- S.D., n = 6) of pregnancy. During day 6, a significant decrease was noted in both the implantation site and the nonimplantation uterine tissue. Microsomal calcium extractable peroxidase showed a similar pattern, with lower specific activity than, the soluble peroxidase. During estrus, the uterine tissue showed the highest activity of calcium-extracted peroxidase (8.7 +/- 1.35 U/mg protein), statistically greater when compared with days 3, 4, 5 and 6 of pregnancy. In conclusion, high peroxidase activity was associated with uterine receptivity. The decrease of activity on day 6 might be due to a progesterone-estrogen interaction, and consequently, hydrogen peroxide can be utilized for hydroxile production by means of the Fenton reaction. Lipoperoxidation may be necessary for changes in membrane fluidity for embryo attachment to endometrial epithelium. PMID- 10687960 TI - Barometers and bladders: a primer on pressures. AB - PURPOSE: We develop a "consilient" (unified) view of pressure as a physical phenomenon and "clinimetric" tool, making a connection between barometers and bladders. MATERIALS AND METHODS: The philosophy, physics and clinical applications of pressure during the last 2 millennia were examined from Lucretius to the modern medical subspecialties. RESULTS: A variety of units and systems of pressure quantification developed as the physics of pressure became understood. Applications of pressure in clinical medicine with distinct physiological relevance have been created for organ systems across the subspecialties. Some measurements have become useful for management of urinary tract and other diseases. CONCLUSIONS: Despite a broad range of units, systems and applications, a consilient view of pressure in medicine can be approached. This perspective is fundamental to understanding the significance of pressures in the expanding clinimetric arena and should mitigate against misplaced concreteness that is tempting in modern medical practice, whereby laboratory tests become virtual realities and are mistaken for patients. PMID- 10687961 TI - Andropause: a misnomer for a true clinical entity. AB - PURPOSE: A progressive decrease in androgen production is common in males after middle age. The resulting clinical picture has been erroneously named male menopause or andropause. A more appropriate designation is androgen decline in the aging male (ADAM). The syndrome is characterized by alterations in the physical and intellectual domains that correlate with and can be corrected by manipulation of the androgen milieu. We review the epidemiological aspects of aging and endocrinological manifestations of ADAM, and provide recommendations for treatment and monitoring of these patients. MATERIALS AND METHODS: We performed MEDLINE, Pubmed, Current Contents and Pharmaceutical Abstracts searches of relevant peer reviewed publications on andropause, male climacteric, adult hypogonadism and aging. In addition, conference proceedings were researched to provide a more complete review of the literature. Information was scrutinized and collated, and contributory data were reviewed and summarized. RESULTS: ADAM is a clinical entity characterized biochemically by a decrease not only in serum androgen, but also in other hormones, such as growth hormone, melatonin and dehydroepiandrosterone. Clinical manifestations include fatigue, depression, decreased libido, erectile dysfunction, and alterations in mood and cognition. CONCLUSIONS: The onset of ADAM is unpredictable and its manifestations are subtle and variable, which has led to a paucity of interest in its diagnosis and treatment. Urological practice commonly includes a large proportion of men older than 50 years. Therefore, it is important for urologists to recognize the manifestations of and be familiar with evaluations necessary to document ADAM as well as its treatment and monitoring. PMID- 10687962 TI - The lion of the union: the pelvic wound of Joshua Lawrence Chamberlain. AB - PURPOSE: Major General Joshua Lawrence Chamberlain is a true American hero. His medical history and war wounds provide a rare snapshot of Civil War era medicine. In particular the most devastating injury was a rifle shot through the pelvis rupturing the bladder and urethra. We describe this injury and how it affected his life to provide insight into late 19th century urological care. MATERIALS AND METHODS: All available references, including biographies, letters, surgical reports, military documents and prior medical summaries, were reviewed regarding Chamberlain's urological history. RESULTS: While leading the Union charge to Petersburg, Virginia on June 18, 1864, Chamberlain was struck with a minie ball anteriorly below the right greater trochanter. The ball coursed obliquely upward disrupting the bladder and urethra, and embedded behind the left acetabulum. An unprecedented wound exploration in the field hospital was performed to extract the bullet and "reconnect severed urinary organs." Hope for recovery was nonexistent as urine was seen exiting the lower wound postoperatively. This genitourinary injury required 4 subsequent repairs during Chamberlain's lifetime and ultimately left him with a draining urethrocutaneous fistula at the penoscrotal junction. CONCLUSIONS: Survival from catastrophic Civil War wounds was rare, especially from "gut wounds" which had a mortality rate of greater than 90%. Chamberlain not only survived but thrived with his sense of duty carrying him back to the battlefield and beyond. He was plagued during his life with recurrent cystitis and epididymo-orchitis, which in an era without antibiotics was especially miserable. Urosepsis is listed as the cause of death on his death certificate and whether this was true is debatable. However, even if this wound did not cause his death, it surely contributed to it. PMID- 10687963 TI - Outpatient adrenalectomy. AB - PURPOSE: To our knowledge we report the initial experience with outpatient, same day laparoscopic adrenalectomy. MATERIALS AND METHODS: Nine select patients were entered into our ambulatory adrenalectomy protocol. Each patient fulfilled certain preoperative and postoperative inclusion criteria, including informed consent, age 70 years or older, body mass index 40 or less, adrenal tumor less than 5 cm., no pheochromocytoma, uncomplicated laparoscopic surgery that was completed by 12 p.m., perioperative hemodynamic stability and pain control by oral analgesics. RESULTS: All 9 patients successfully underwent outpatient laparoscopic adrenalectomy. Average patient age was 53 years and average adrenal tumor size was 2 cm. Mean surgical time was 2.3 hours and mean blood loss was 53 ml. The diagnosis was aldosteroma in 7 cases, enlarging adenoma in 1 and myelolipoma in 1. Average postoperative hospital stay was 416 minutes (range 300 to 570). Postoperative analgesia comprised 6 mg. morphine sulfate and 32 mg. ketorolac. The only complication was a local abscess requiring delayed drainage at 2 weeks. No other patient was rehospitalized for any reason. A followup questionnaire survey revealed excellent patient satisfaction. CONCLUSIONS: To our knowledge we report the initial series of outpatient laparoscopic excision of a solid organ, the adrenal gland. Ambulatory adrenalectomy is feasible and safe, and results in high patient satisfaction. However, ambulatory adrenalectomy should be restricted to highly select patients and performed by minimally invasive surgeons who have considerable experience with laparoscopic adrenal surgery. PMID- 10687964 TI - Variation in clinical outcome following shock wave lithotripsy. AB - PURPOSE: We measure and compare operator specific success rates of extracorporeal shock wave lithotripsy (ESWL) performed by 12 urologists in 1 unit to determine interoperator variation. MATERIALS AND METHODS: From January 1, 1994 to September 1, 1997 a total of 5,769 renal and ureteral stones received 9,607 ESWL treatments by 15 urologists with a Dornier MFL 5000 lithotriptor. The 3-month followup data are available for 4,409 stones. Outcome measures consisted of patient demographics, stone characteristics, technical details of lithotripsy, and stone free and success rates by treating urologists. RESULTS: Treatment results were analyzed for 12 urologists (surgeons A to L) who treated more than 100 stones each, totaling 4,244 with followup information available. Mean stone-free and success rates were 50.6% and 72.3%, respectively. Surgeon A had significantly higher stone-free and success rates of 56.2% and 76.7%, respectively (p<0.05), with treatment results from 877 stones, which was a significantly higher number than others (p<0.05). Significant differences existed in mean number of shocks delivered among urologists (p = 0.0001), with surgeons A and J delivering the highest mean numbers (2,317 and 2,801, respectively). There was no difference in treatment duration (p = 0.75) but variation existed among urologists in terms of mean maximum treatment voltage (p = 0.0001). Mean fluoroscopy time at 4.1 minutes was higher for surgeon A than others (p<0.05). Mean complication rate following ESWL was 4.9% with no difference among urologists (p = 0.175). Re-treatment was required in 21.7% of cases and surgeon A had the lowest rate (15.9%, p<0.05). CONCLUSIONS: We demonstrated clinically and statistically significant intra institutional differences in success rates following ESWL. The best results were obtained by the urologist who treated the greatest number of patients, used a high number of shocks and had the longest fluoroscopy time. Accurate targeting is crucial when using a lithotriptor, such as the Dornier MFL 5000, with a narrow focal zone of 6.5 mm. in diameter. Other centers should be encouraged to develop similar programs of outcome analysis in an attempt to improve performance. PMID- 10687965 TI - Nephrolithiasis associated with autosomal dominant polycystic kidney disease: contemporary urological management. AB - PURPOSE: We evaluate the role of contemporary urological intervention in patients with nephrolithiasis associated with autosomal dominant polycystic kidney disease. MATERIALS AND METHODS: Intervention for upper tract stones associated with autosomal dominant polycystic kidney disease was performed in 5 women and 2 men 29 to 65 years old (mean age 47). Indications for intervention consisted of flank pain in 6 patients and/or hematuria in 2. A total of 12 procedures (mean 1.7 per patient) were performed, including shock wave lithotripsy in 6 patients, percutaneous nephrolithotomy in 2, retrograde endoscopy or manipulation in 3 and extended pyelonephrolithotomy in 1. RESULTS: All patients were rendered stone free or had only residual "dust." Hospital stay for 5 patients was 1 night or less and there were no complications. Renal function for each patient was stable or improved as measured by serum creatinine. CONCLUSIONS: Most patients with autosomal dominant polycystic kidney disease who require intervention for nephrolithiasis can be safely and effectively treated with essentially any or all contemporary, minimally invasive techniques. The choice of intervention can be based primarily on size and location of the upper tract stones rather than the associated presence of polycystic kidneys. PMID- 10687966 TI - Surgical management of renal tumors 4 cm. or less in a contemporary cohort. AB - PURPOSE: We evaluated a patient cohort with renal tumors 4 cm. or less treated with partial or radical nephrectomy. We compared patient and tumor characteristics, and survival in these 2 groups. MATERIALS AND METHODS: We retrospectively analyzed the records of 670 patients with a median age of 63 years treated surgically for renal cell carcinoma between July 31, 1989 and July 31, 1997. Renal tumors 4.0 cm. or less were noted in 252 patients (38%) who underwent a total of 262 procedures, including 183 radical (70%) and 79 partial (30%) nephrectomies. Ten patients required 2 operations each because of bilateral renal cell carcinoma. Median followup was 40 months. We compared clinicopathological parameters in the partial and radical nephrectomy groups using chi-square or Wilcoxon analysis as appropriate. Survival analysis was determined by the log rank test and Cox regression model. RESULTS: The partial and radical nephrectomy groups were comparable with respect to gender ratio, tumor presentation, histological classification, pathological stage and complication rate. Median tumor size was 2.5 and 3.0 cm. in the partial and radical nephrectomy groups, respectively (p = 0.0001). Resection was incomplete in 1 patient (1.3%) in the partial and none in the radical nephrectomy group. There was no local recurrence after either procedure, and no significant difference in disease specific, disease-free and overall survival (p = 0.98, 0.23 and 0.20, respectively). CONCLUSIONS: Patients with a small renal tumor have similar perioperative morbidity, pathological stage and outcome regardless of treatment with partial or radical nephrectomy. Therefore, partial nephrectomy remains a safe alternative for tumors of this size. PMID- 10687967 TI - Nephron sparing surgery for central renal tumors: experience with 33 cases. AB - PURPOSE: Nephron sparing surgery is standard treatment for small, peripherally located renal cell carcinoma. In patients with a solitary kidney, bilateral tumors or impaired renal function nephron sparing surgery provides the only option to nephrectomy and subsequent hemodialysis or transplantation. We retrospectively investigated the value of nephron sparing surgery for centrally located renal cell carcinoma. MATERIALS AND METHODS: Between 1969 and 1997, 311 renal tumor enucleations were performed at our institution. The tumor was centrally located in 33 cases. The indication for enucleation was elective in 7 cases and imperative in 26, including bilateral tumor in 16 (metachronous in 9 and synchronous in 7), chronic renal failure in 4 and solitary kidney in 6. Four patients had metastasis at enucleation. RESULTS: Convalescence was unremarkable in 28 cases. Hemorrhage occurred in 1 patient, a urinary fistula in 2 and a local abscess secondary to a urinary fistula in 1. One patient died postoperatively of heart failure. Average serum creatinine was 1.25, 1.63 and 1.33 mg./dl. preoperatively, at hospital discharge and at a mean followup of 33 months, respectively. Hemodialysis was necessary transiently during convalescence in 1 patient and permanently starting 6 years after enucleation in another. Definitive histology revealed oncocytoma in 4 cases and renal cell carcinoma in 29. Disease was stages pT1 to pT3 in 9, 18 and 2 cases, and grades 1 to 3 in 6, 18 and 5, respectively. Local recurrence developed in 2 patients. Mean followup was 5.2 years (range 0.3 to 16.7). At a mean followup of 6.2 years (range 0.7 to 16.7) 20 patients were free of disease. In addition to the patient who died postoperatively, 9 died of renal cell carcinoma at a mean of 1.6 years (range 0.3 to 5.3) and 3 died of other causes at 5, 11 and 12 years postoperatively, respectively. No patient who underwent elective enucleation died. CONCLUSIONS: Nephron sparing surgery for centrally located kidney tumors is technically feasible and associated with an acceptable complication rate. Local tumor control is excellent, and the overall prognosis depends on contralateral disease and metastasis. Benign tumors may be diagnosed and removed without loss of the kidney. By avoiding hemodialysis quality of life is improved. PMID- 10687968 TI - Management of superficial transitional cell carcinoma in the intramural ureter: what to do? AB - PURPOSE: We analyze the evolution of superficial transitional cell carcinoma in the intramural distal ureter treated with transurethral resection. MATERIALS AND METHODS: A total of 19 patients underwent transurethral resection of the intramural distal ureter with a mean followup of 57 months. All cases were diagnosed as superficial transitional cell carcinoma and all but 2 had a history of bladder tumor. Upper urinary tract followup consisted of excretory urography every 6 months and ureterorenoscopy in cases with a doubtful diagnosis or positive cytology. RESULTS: Pathological examination revealed stage Ta disease in 42%, T1 in 31.5% and Tx in 26.3% of intramural tumors. Upper urinary tract recurrence was noted in 8 patients (42.1%), including 5 (62.5%) with involvement of the distal ureter. Nontumoral stenosis of the distal ureter in 3 cases was treated endoscopically. An endoscopic procedure resolved 75% of recurrences. A high surgical risk patient who did not undergo open surgery died of recurrence. CONCLUSIONS: Superficial transitional cell carcinoma of the intramural ureter is uncommon in the setting of multiple bladder tumors and recurrent bladder carcinoma. There was a 42.1% rate of ipsilateral recurrence and endoscopic treatment allowed us to preserve 89.5% of the involved renal units. Closer followup of the urinary tract must be performed since these tumors have a higher incidence of upper urinary tract recurrence. PMID- 10687969 TI - Clinical and pathological characteristics of micropapillary transitional cell carcinoma: a highly aggressive variant. AB - PURPOSE: We present preliminary clinical, histochemical and molecular findings for 5 patients with micropapillary transitional cell carcinoma of the bladder, a rare histological variant not widely recognized in the urological literature. MATERIALS AND METHODS: The 5 patients were prospectively identified. In 3 cases immunohistochemical staining for expression of CD31, p53, E-cadherin, and alpha, beta and gamma-catenin was performed on paraffin embedded tissue. Sequencing was used to identify point mutations in exons 5 to 9 of p53, and exons 1 and 2 of H ras. RESULTS: Of the patients 2 died within 1 year of presentation to our institution with rapid local extension along the bladder serosal surface and ureteral sheaths. Another patient had progression to invasive disease within 22 months. In the 3 cases with immunohistochemical staining p53 was negative, despite positive staining of nonmicropapillary transitional cell carcinoma within the same specimen. Stains for the angiotrophic marker CD31 were negative. In all 3 cases normal membrane associated alpha, beta and gamma-catenin expression was present. Examination of p53 sequences revealed a single point mutation in exon 8 of 1 case. In 2 cases different mutations in exon 1 of H-ras were noted. CONCLUSIONS: Micropapillary transitional cell carcinoma is a rare and highly aggressive variant. Paradoxically, our study demonstrated no significant p53 abnormalities. The lacunar histological pattern did not appear to represent invasion of vascular spaces. Rather, these tumors seemed to have the ability to disrupt and replace the normal stromal matrix to achieve rapid nonendothelial extension. Thus, micropapillary histology may predict a lesser likelihood of surgical cure. PMID- 10687970 TI - Comparison of molecular and conventional strategies for followup of superficial bladder cancer using decision analysis. AB - PURPOSE: Patients with superficial bladder cancer require long-term surveillance for recurrence. We compared the cost of cystoscopy and cytology (standard care) to that of urinary markers (modified care) for patients with a history of superficial bladder cancer. MATERIALS AND METHODS: We constructed a decision analysis model that compared the 2 strategies for a hypothetical followup interval of 3 years. Probabilities required for the decision tree were based on a cohort of 361 patients diagnosed with superficial bladder cancer from 1987 to 1997. Sensitivity analyses were used to determine whether test sensitivity and specificity would affect cost thresholds. Costs for each strategy were then applied to actual practice patterns. RESULTS: The cost of modified care ranged from $158 to $228 for each followup visit when using a urinary marker with a sensitivity and specificity of 95% and 77%, respectively. The cost of standard care was $240 for each followup visit. Based on sensitivity analyses the probability of disease recurrence and urinary marker accuracy were important determinants of expected costs. Mean number of followup assessments for patients followed more than 3 years was 4.3, 2.2 and 1.5 for years 1, 2 and 3, respectively. Cumulative costs of modified care were lower than those of standard care. CONCLUSIONS: Urinary marker testing for followup of patients with superficial bladder cancer is less expensive than the standard method of cystoscopy and urinary cytology based on our model. Future studies will be required to consider other factors that could affect the cost advantage of urinary markers, including indirect costs, the psychosocial impact of testing and different surveillance frequencies. PMID- 10687971 TI - The role of bcl-2, p53, and ki-67 index in predicting tumor recurrence for low grade superficial transitional cell bladder carcinoma. AB - PURPOSE: We assess the prognostic significance of bcl-2 expression, p53 mutation and ki-67 index for low grade, superficial transitional cell bladder carcinoma. MATERIALS AND METHODS: The medical records of 93 cases of primary, low grade (24 G1, 69 G2), superficial (70 pTa, 23 pT1) transitional cell carcinoma of the bladder were reviewed. Association of bcl-2, p53 and ki-67 index immunoreactivity with tumor grade and stage was examined. Prognostic significance of tumor grade, pathological stage, bcl-2 expression, p53 mutation and ki-67 index in predicting tumor recurrence was assessed. RESULTS: Of the tumors 60 (70%) had p53 mutation and 9 (10.5%) expressed bcl-2. These 2 markers did not relate to tumor grade or pathological stage. Median ki-67 index was 10.9% and positively correlated with tumor grade. Recurrence was noted in 34.9% of patients with a median followup of 26 months (range 1 to 84). The ki-67 index was the only significant prognostic indicator in univariate and multivariate analyses. This marker can further distinguish grade 2 tumors with a favorable prognosis from those with an unfavorable outcome. CONCLUSIONS: The ki-67 labeling index is an independent predictor of tumor recurrence for patients with primary superficial, low grade bladder cancers. PMID- 10687972 TI - Efficacy and safety of valrubicin for the treatment of Bacillus Calmette-Guerin refractory carcinoma in situ of the bladder. The Valrubicin Study Group. AB - PURPOSE: We assess the efficacy and safety of intravesical valrubicin for the treatment of carcinoma in situ in patients with failure or recurrence after bacillus Calmette-Guerin (BCG) and who otherwise would have undergone cystectomy. Total anthracycline recovery in urine samples obtained within 24 hours of valrubicin administration was assessed in a subset of patients. MATERIALS AND METHODS: A total of 90 patients with recurrent carcinoma in situ after failed multiple prior courses of intravesical therapy, including at least 1 course of BCG, participated in this open label, noncomparative study. Each patient received 6 weekly instillations of 800 mg. intravesical valrubicin. Disease evaluations were made at baseline and 3-month intervals following treatment. Evaluations included cystoscopy with biopsy and urine cytology. Toxicity was noted throughout treatment and followup. No evidence of disease recurrence for 6 months or greater was considered a complete response. RESULTS: Of 90 patients 19 (21%) had a complete response, including 7 who remained disease-free at the last evaluation, with a median followup of 30 months. Additionally, 14 patients who did not meet the strict protocol definition of complete response had superficial Ta disease only. Median time to failure and/or last followup for complete responders was greater than 18 months. Recurrence has been noted in 79 patients to date, including only 2 with clinically advanced disease (stage T2). Of these 79 patients 44 (56%, 4 responders and 40 nonresponders) underwent radical cystectomy. Of the 41 patients with known pathological stage 6 (15%) had stage pT3 or greater at cystectomy. Four patients died of bladder cancer during the median followup of 30 months, none of whom was a complete responder or underwent cystectomy following valrubicin. The main side effects of valrubicin therapy were reversible local bladder symptoms. CONCLUSIONS: Valrubicin was effective and well tolerated in patients with carcinoma in situ of the bladder refractory to BCG therapy. Delaying cystectomy while attempting salvage therapy with valrubicin does not pose an undue risk to most patients. PMID- 10687973 TI - Effect of bladder management on urological complications in spinal cord injured patients. AB - PURPOSE: The optimal method of bladder management in spinal cord injured patients remains controversial. We investigated the association of type of bladder management with urological complications in these patients. MATERIALS AND METHODS: We retrospectively reviewed the medical records, upper tract imaging and video urodynamics of 316 posttraumatic spinal cord injured patients. Mean followup plus or minus standard deviation since injury was 18.3+/-12.4 years. Patients were categorized according to bladder management method, including chronic urethral catheterization, clean intermittent catheterization, spontaneous voiding and suprapubic catheterization in 114, 92, 74 and 36, respectively. No significant differences in patient age at injury, followup interval, or level, completeness or mechanism of injury were noted among bladder management method groups. Infection, stone disease, urethral complications and radiographic abnormalities were recorded. RESULTS: Of the 398 complications recorded 236 developed in 61 (53.5%) patients on chronic urethral catheterization, 57 in 25 (27.2%) on clean intermittent catheterization, 57 in 24 (32.4%) who voided spontaneously and 48 in 16 (44.4%) on suprapubic catheterization. The intermittent catheterization group had statistically significant lower complication rates compared with the urethral catheterization group and no significantly higher complication rates relative to all other management methods for each type of complication studied. The percent of patients with complications was greater in the chronic urethral catheterization group only 5 years after injury, while the percent in all other management groups remained similar up to 15 years after injury. CONCLUSIONS: Clean intermittent catheterization is the safest bladder management method for spinal cord injured patients in terms of urological complications. Inappropriate selection of a bladder management method not only adversely affects patient quality of life, but also has a significant detrimental impact on the economic status of the health care system. PMID- 10687974 TI - Removal of UroLume endoprosthesis: experience of the North American Study Group for detrusor-sphincter dyssynergia application. AB - PURPOSE: We present the experience of the North American UroLume Multicenter Study Group with removal of the UroLume endoprosthesis. MATERIALS AND METHODS: A total of 160 neurologically impaired patients were enrolled in the North American UroLume Multicenter Study Group for detrusor external sphincter dyssynergia application. Analysis was performed in 2 groups of patients in which the device was removed during insertion and after implantation, respectively. RESULTS: Device retrieval was required during insertion in 21 patients (13%) mainly due to misplacement or migration in 17. Extraction was done with minimal complications and in all but 2 cases subsequent UroLume implantation was successful. Of 158 men with the device in place 31 (19.6%) required removal. In 34 procedures 44 devices were removed, mainly due to migration. Time from implantation to removal ranged from 4 days to 66 months (mean 22 months). The UroLume was removed en bloc in 20 cases and in parts or wire by wire in 19. The majority of patients had no or minimal complications after extraction. Only 2 patients had serious temporary complications, including bleeding and urethral injury, with no lasting consequences. No malignancy developed as a result of UroLume insertion. CONCLUSIONS: While there is a potential for urethral injury and bleeding, UroLume endoprosthesis removal is largely a simple procedure with minimal complications and consequences. PMID- 10687975 TI - Nocturia in the adult: classification on the basis of largest voided volume and nocturnal urine production. AB - PURPOSE: We propose a criterion to clarify the underlying etiologies of nocturia. MATERIALS AND METHODS: Frequency-volume charts were recorded for 24 hours by 35 men and 32 women who were subjectively free of lower urinary tract symptoms and had no evidence of voiding disorders. At least 5 subjects were included in each of 5 age groups of 20 to 49, 50 to 59, 60 to 69, 70 to 79, 80 to 89 years for both genders. The charts were used to make a tentative criterion, which was validated in 39 elderly individuals with nocturnal frequency. RESULTS: Correlation and regression analyses indicated that the quotients of nocturnal urine output divided by body weight (U(N)/BW) and largest voided volume divided by body weight (V(L)/BW) were useful classification factors. Cutoffs, which were set close to worst quartiles, were 10 ml./kg. for U(N)/BW and 4 ml./kg. for V(L)/BW, respectively. Correction by body weight made the same criterion applicable irrespective of weight. Symptomatic elderly patients were classified into 3 mutually exclusive groups of nocturnal polyuria (U(N)/BW greater than 10 ml./kg.), low bladder capacity (V(L)/BW less than 4 ml./kg.) and combined nocturia. Of 20 subjects who voided 2 times a night 11 (55%) were classified as having nocturnal polyuria and 15 of 19 who voided 3 or 4 times (79%) were classified as having low bladder capacity or combined nocturia. CONCLUSIONS: The criterion provides a reasonable distinction of etiologies of nocturia, and may be usefulness in examination and treatment. PMID- 10687976 TI - Cowper's syringocele: symptoms, classification and treatment of an unappreciated problem. AB - PURPOSE: Cowper's syringocele is a rare deformity in the male urethra that is a distention of the duct of the bulbourethral (Cowper's) gland. We report on 7 cases, review the symptoms and pathophysiology, and propose a simplified classification of this uncommon lesion. MATERIALS AND METHODS: We reviewed 7 cases of Cowper's syringocele diagnosed from 1997 to 1998 at our hospital. RESULTS: Cowper's syringocele was diagnosed in 7 patients 25 to 51 years old with persistent post-void dribbling, frequency, urethral pain, hematuria or sudden urethral discharge. Diagnosis was made with urethrocystoscopy or retrograde urethrogram. Cowper's syringocele may be closed (a distended cyst-like swelling in the wall of the urethra) or open (an opening enabling urine reflux into the syringocele). In 2 patients asymptomatic open syringocele was diagnosed. In 1 patient symptomatic syringocele resolved spontaneously following an infection. In 4 patients open syringocele was treated with transurethral marsupialization because of persistent post-void dribbling. Postoperatively patients were completely symptom-free with a mean followup of 12 months (range 6 to 23). CONCLUSIONS: Cowper's syringocele may be more common than currently realized. Urologists should rule out this possibility in young male patients with lower urinary tract symptoms and persistent post-void dribbling as it can be treated easily. PMID- 10687977 TI - Intraurethral application of alprostadil in patients with failed inflatable penile prosthesis. AB - PURPOSE: Many men who underwent penile prosthesis implantation before the advent of oral and injection therapy present for replacement of a malfunctioning prosthesis but choose not to undergo revision surgery because of personal, medical or reimbursement issues. Others with normally functioning prostheses report significant difficulties with "cold glans," and they and their partners observe decreased engorgement and temperature of the glans penis with the inflated penile prosthesis, despite adequate stimulation. Intracorporal injection therapy is contraindicated in any patient with a penile prosthesis and use of a vacuum erection device may result in prosthesis cylinder rupture. In these patients intraurethral application of alprostadil may restore prosthesis function and permit satisfactory intercourse. We evaluate the efficacy of a medicated urethral system for erection (MUSE) using alprostadil to restore function for men with a failed prosthesis, and improve glans penis temperature sensation and engorgement for those with a functioning prosthesis. MATERIALS AND METHODS: From February 1997 to February 1998, 28 men 47 to 81 years old (mean age 61.2) with a penile prosthesis were treated with alprostadil. Of the patients 11 had penile prosthesis failure (group 1) and 17 reported decreased glans penis engorgement (group 2). In 18 cases erections were observed at the clinic. Doses of alprostadil varied from 250 to 1,000 microgm. (mean 566). RESULTS: Of the 28 patients 23 had a response to alprostadil. Erections were sufficient for intercourse in 7 of 11 group 1 patients, and 10 of 17 group 2 were satisfied with treatment. There was no device specific morbidity but 12 men discontinued use of alprostadil because of penile pain. A significant or excellent response was noted in 10 of 18 men observed at the clinic. CONCLUSIONS: Intraurethral alprostadil may be used to restore or improve function of a penile prosthesis in patients with a malfunctioning device or lack of glans penis engorgement, with low expected morbidity. PMID- 10687978 TI - Sexual function in men with diabetes type 2: association with glycemic control. AB - PURPOSE: We evaluated the association of glycemic control with erectile dysfunction in men with diabetes type 2. MATERIALS AND METHODS: A convenience sample of men with diabetes type 2 at the Cleveland Veterans Affairs Medical Center completed questions 1 to 5 of the International Index of Erectile Function. The primary outcome measure was erectile function score, calculated as the sum of questions 1 to 5. Details of disease duration, complications, medication use, patient age and level of glycosylated hemoglobin were obtained by reviewing the medical record. RESULTS: Mean subject age plus or minus standard deviation was 62.0+/-12.3 years, mean hemoglobin A1c was 8.1%+/-1.9% and mean erectile function score was 16.6+/-5.9 (range 5 to 23). Stratified analysis revealed that mean erectile function score decreased as hemoglobin A1c increased (analysis of variance p = 0.002). The test for linearity was also significant (p = 0.001). There were no statistically significant associations of levels of glycemic control with alpha-blocker, beta-blocker or diuretic use. Bivariate analysis showed a significant correlation of hemoglobin A1c with neuropathy but not with patient age, duration of diabetes, alpha-blockers, beta-blockers or diuretics. Multivariate analysis demonstrated that hemoglobin A1c was an independent predictor of erectile function score (p<0.001) even after adjusting for peripheral neuropathy, which was also an independent predictor (p = 0.023). CONCLUSIONS: Our data add to the growing body of literature suggesting that erectile dysfunction correlates with the level of glycemic control. Peripheral neuropathy and hemoglobin A1c but not patient age were independent predictors of erectile dysfunction. PMID- 10687979 TI - Testis biopsy findings in the spinal cord injured patient. AB - PURPOSE: Azoospermia after electroejaculation in spinal cord injured men may be due to testicular failure or obstruction. These men can initiate pregnancy with assisted reproductive techniques, such as intracytoplasmic sperm injection, but only if sperm are present in the testis biopsy. We analyzed the histopathology of testis biopsies from spinal cord injured men and assessed whether patient factors were predictive of testis biopsy pathology. MATERIALS AND METHODS: A total of 50 paraplegic men undergoing testis biopsy were divided into 2 groups based on normal or abnormal testis histopathology. Patient age, post-injury years, level of lesion, hormonal status and semen analysis results were compared. RESULTS: Spermatogenesis was normal in 28 of the 50 patients. Hypospermatogenesis was exhibited in 15, maturation arrest at the spermatid stage in 6 and maturation arrest at the spermatocyte stage in 1 of the 22 abnormal cases. Nevertheless, mature sperm were identified in 43 of 50 biopsies (normal spermatogenesis and hypospermatogenesis). Men with normal spermatogenesis had better forward progression of sperm and a higher testosterone-to-luteinizing hormone ratio. Otherwise, there was no statistically significant correlation between study variables and testis biopsy results. No factors were predictive of testis biopsy histopathology. CONCLUSIONS: The documentation of mature sperm in 43 of 50 biopsies from spinal cord injured patients suggests that a high rate of sperm retrieval is possible using testicular sperm extraction if sperm cannot be retrieved from the ejaculate. With intracytoplasmic sperm injection techniques the majority of spinal cord injured men retain fertility potential, even if azoospermic following electroejaculation. PMID- 10687980 TI - Recognizing abnormal marker results that do not reflect disease in patients with germ cell tumors. AB - PURPOSE: The judicious use of serum alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG) and lactate dehydrogenase is key to appropriate management of patients with germ cell tumors. Elevated AFP and/or HCG generally indicate active disease. We describe patients with germ cell tumors who had elevated serum AFP and/or HCG but no active disease, despite careful repeat evaluation. MATERIALS AND METHODS: Histories of 6 cases of germ cell tumors that remained in remission despite abnormal serum AFP and/or HCG were reviewed. RESULTS: Markers were only modestly elevated, remained constant or spontaneously normalized during repeat measurements, and there was no other clinical or radiographic evidence of disease. Patients were treated conservatively with physical examination, radiological tests and repeat marker assays, with no relapse to date. CONCLUSIONS: Stable, low increases in serum AFP and HCG may not represent active disease. Careful repeat evaluation will determine whether the markers increase. If no change is noted after appropriate studies have been reviewed by an experienced practitioner to exclude active disease from diagnosis, then consideration should be given to managing such cases with close surveillance to avoid unnecessary chemotherapy. PMID- 10687981 TI - Dramatic suppression of plasma and urinary prostate specific antigen and human glandular kallikrein by antiandrogens in male-to-female transsexuals. AB - PURPOSE: Prostate specific antigen (PSA) and human glandular kallikrein (hK2) are mainly produced by the prostate and their genes are regulated by androgens through the androgen receptor. We determine whether PSA and hK2 change significantly in plasma and urine after antiandrogen treatment in male-to-female transsexuals. MATERIALS AND METHODS: Plasma and urine PSA and hK2 were measured with highly sensitive immunofluorometric procedures capable of detecting within 1 or 6 ng./l. PSA or hK2, respectively. Study groups consisted of 10 men treated with cyproterone acetate only (group 1), 15 transdermal estradiol plus cyproterone acetate (group 2) and 31 ethinyl estradiol plus cyproterone acetate (group 3). Plasma and urine samples were collected before initiation of treatment as well as after 4 months of hormonal therapy. For a subset of group 3 patients blood and urine samples were also obtained after 12 months of treatment. RESULTS: Cyproterone acetate, a steroidal antiandrogen, alone or with estradiol was able to suppress greater than 90% of plasma and urinary PSA and hK2 concentration after 4 or 12 months of therapy. CONCLUSIONS: Cyproterone acetate therapy causes dramatic suppression of plasma and urinary PSA and hK2 in men without prostate cancer. Since cyproterone acetate is used for prostate cancer treatment, suppression of PSA after hormonal therapy may not accurately reflect therapy success in reducing tumor burden. PMID- 10687982 TI - Prostate cancer detection at low prostate specific antigen. AB - PURPOSE: At low prostate specific antigen (PSA) the indication for prostate biopsy is usually an abnormal digital rectal examination. We evaluate the diagnostic value of PSA, digital rectal examination, transrectal ultrasonography and tumor characteristics at low PSA (0 to 4.0 ng./ml.). We confirm and add to recent evidence that digital rectal examination has a low predictive value and that many significant cancers at this PSA range may be missed. MATERIALS AND METHODS: From 1994 to 1997 a total of 10,523 participants 54 to 74 years old were randomized to screening in the Rotterdam section of the European Randomized Study of Screening for Prostate Cancer. Of the participants 9,211 (87.5%) had PSA less than 4.0 ng./ml., and underwent digital rectal examination and transrectal ultrasonography. Expected rates of prostate cancer detection were calculated using logistic regression analysis. Radical prostatectomy was performed in about half of the 478 men diagnosed with prostate cancer. Tumors were characterized by pT category, Gleason score and cancer volume in 166 processed radical prostatectomy specimens. In 50 of these cases PSA was 0 to 4.0 ng./ml. RESULTS: The positive predictive value of digital rectal examination and transrectal ultrasonography at PSA 0 to 4.0 ng./ml. was only 9.7%. Positive predictive value strongly depended on PSA. Sensitivity was calculated by using estimates of the prevalence of sextant biopsy detectable prostate cancers. Of 760 detectable cancers 478 (67%) were diagnosed irrespective of PSA in men screened with digital rectal examination, transrectal ultrasonography and PSA. Only 127 of 348 detectable prostate cancers (36.5%) were actually diagnosed in men with PSA 2 to 4 mg./ml. The importance of these missed cancers was evaluated with parameters of tumor aggressiveness within PSA ranges. CONCLUSIONS: Approximately half of the tumors missed with PSA 0 to 4 ng./ml. had aggressive characteristics (Gleason score 7 or greater, Gleason 4-5 components) and were organ confined. These tumors should be diagnosed and treated according to the present understanding of their natural history. More sensitive and selective screening strategies are needed. Presently a wrong "window of opportunity" is used for early detection of prostate cancer. PMID- 10687983 TI - Predictors of first repeat biopsy cancer detection with suspected local stage prostate cancer. AB - PURPOSE: We determine demographic and tumor related predictors of repeat biopsy cancer detection in men with suspected stage T1c-2 prostate cancer. MATERIALS AND METHODS: The study population included 298 consecutive men with suspected stage T1c-2 prostate cancer who had a benign prostate biopsy at 1 institution between January 1, 1992 and April 1, 1999 and underwent 1 repeat biopsy. Mean age plus or minus standard deviation was 66.8+/-6.7 years for 133 black (55%) and 165 white (45%) patients. Clinical measures included determination of high grade prostatic intraepithelial neoplasia in benign biopsy specimens, Gleason score of malignant biopsy specimens, prostate specific antigen (PSA), PSA density, annualized interbiopsy PSA change, percent free PSA (201 cases) and PSA velocity (171). RESULTS: Cancer was detected on repeat biopsy in 80 cases (27%). Significant differences between patients with benign and malignant repeat biopsies included age (p = 0.001), PSA density (p = 0.0001), percent free PSA (p = 0.0001) and PSA velocity (p = 0.009). High grade prostatic intraepithelial neoplasia in an initial benign biopsy was not predictive of cancer in repeat biopsy (p = 0.12). Multiple logistic regression analysis of all cases showed that age (p = 0.002) and PSA density (p = 0.0002) were independent predictors of cancer. Subset multiple logistic regression analysis modeled with age, PSA density and percent free PSA demonstrated that age (p = 0.002) and percent free PSA (p = 0.0001) were significant independent predictors of malignancy. Subset multiple logistic regression analysis modeled with age, PSA density, percent free PSA and PSA velocity revealed that age (p = 0.02) and percent free PSA (p = 0.0003) were significant independent predictors of cancer. There were no significant differences between the Gleason scores of cancers detected on repeat biopsy compared to 587 stage T1c-2 cancers detected on initial biopsy during the study period (p = 0.09). PSA, PSA density, percent free PSA and PSA velocity were not significantly different among men without a cancer diagnosis who had high grade neoplasia in 1 or 2 benign biopsies. CONCLUSIONS: Greater than 25% of this population of select patients with suspected stage T1c-2 prostate cancer had malignancy detected on repeat biopsy. Percent free PSA was the most powerful predictor of cancer. High grade prostatic intraepithelial neoplasia was not a predictor of repeat biopsy cancer detection and PSA functions were similar among men without cancer who did and did not have high grade neoplasia in 1 or more benign biopsies. This finding suggests that high grade prostatic intraepithelial neoplasia may not be a reliable indicator of clinically significant existing prostate cancer. PMID- 10687984 TI - Strategy for repeat biopsy in patients with high grade prostatic intraepithelial neoplasia. AB - PURPOSE: The finding of high grade prostatic intraepithelial neoplasia in a biopsy specimen without prostate cancer warrants repeat biopsy because of the risk of concurrent cancer. However, to our knowledge the optimal repeat biopsy technique has not yet been defined. We determined the optimal subsequent biopsy strategy for detecting concurrent cancer in patients diagnosed with high grade prostatic intraepithelial neoplasia. MATERIALS AND METHODS: Of 63 men with isolated high grade prostatic intraepithelial neoplasia on initial biopsy 45 underwent repeat biopsy within 1 year. Certain biopsy patterns were used for repeat biopsy, including only the neoplasia site in 8 men, sextant in 12, sextant plus bilateral transition zone in 13 and 11 core multisite directed (sextant, bilateral transition zone, bilateral anterior horn of the peripheral zone and midline peripheral zone) in 12. We compared the location of high grade disease on the initial biopsy with the cancer site on repeat biopsy. RESULTS: Repeat biopsy revealed cancer in 10 of the 45 men (22%), and the sites of high grade prostatic intraepithelial neoplasia and cancer correlated in 6. Cancer was detected at the sextant locations in 9 men. Of the 15 cores positive for cancer 8 were at the original high grade neoplasia site, 6 at a random sextant biopsy site and 1 in the transition zone. High grade disease was discovered bilaterally in 1 man, while prostatic intraepithelial neoplasia and cancer were detected on the same side in the remaining 9. CONCLUSIONS: The optimal repeat biopsy strategy for patients with high grade prostatic intraepithelial neoplasia has not yet been determined but at a minimum it should include targeting the area of known high grade disease and the ipsilateral sextants. PMID- 10687985 TI - Is low serum free testosterone a marker for high grade prostate cancer? AB - PURPOSE: The association of free and total testosterone with prostate cancer is incompletely understood. We investigated the relationship of serum free and total testosterone to the clinical and pathological characteristics of prostate cancer. MATERIALS AND METHODS: We retrospectively reviewed the clinical records of 117 consecutive patients treated by 1 physician and diagnosed with prostate cancer at our medical center between 1994 and 1997. Low free and total testosterone levels were defined as 1.5 or less and 300 ng./dl., respectively. RESULTS: After evaluating all 117 patients we noted no correlation of free and total testosterone with prostate specific antigen, patient age, prostatic volume, percent of positive biopsies, biopsy Gleason score or clinical stage. However, in patients with low versus normal free testosterone there were an increased mean percent of biopsies that showed cancer (43% versus 22%, p = 0.013) and an increased incidence of a biopsy Gleason score of 8 or greater (7 of 64 versus 0 of 48, p = 0.025). Of the 117 patients 57 underwent radical retropubic prostatectomy. In those with low versus normal free testosterone an increased mean percent of biopsies demonstrated cancer (47% versus 28%, p = 0.018). Pathological evaluation revealed stage pT2ab, pT2c, pT3 and pT4 disease, respectively, in 31%, 64%, 8% and 0% of patients with low and in 40%, 40.6%, 12.5% and 6.2% in those with normal free testosterone (p>0.05). CONCLUSIONS: In our study patients with prostate cancer and low free testosterone had more extensive disease. In addition, all men with a biopsy Gleason score of 8 or greater had low serum free testosterone. This finding suggests that low serum free testosterone may be a marker for more aggressive disease. PMID- 10687986 TI - Beyond prostate specific antigen--markers for prostate cancer for the 21st Century. PMID- 10687987 TI - The role of preoperative epoetin alfa in men undergoing radical retropubic prostatectomy. AB - PURPOSE: The safety and effects on hematocrit of recombinant human erythropoietin (epoetin alfa) were evaluated in men undergoing radical retropubic prostatectomy. MATERIALS AND METHODS: Between February 1, 1997 and November 2, 1998, 305 men with clinically localized adenocarcinoma of the prostate underwent radical retropubic prostatectomy performed by a single surgeon (H. L.). Of these men 283 with a baseline hematocrit of less than 48% received 600 IU/kg. epoetin alfa 14 days (-14) and 7 days (-7) before radical retropubic prostatectomy. Hematocrit was measured at baseline on day -14, on day -7, just before anesthesia induction on day 0, immediately postoperatively and on the day of discharge home. The number of allogeneic units transfused, and all intraoperative and postoperative complications were recorded. RESULTS: Mean hematocrit at baseline on day -14 and at induction on day 0 was 42.9% and 45.8%, respectively (p = 0.0001). The frequency of hematocrit decreasing, showing no change or increasing 0.1 to 1.9, 2.0 to 3.9 or greater than 4.0 hematocrit points was 16.5%, 0.5%, 23%, 22% and 38%, respectively. Of the men 17% had no increase in hematocrit. A weak correlation existed between baseline hematocrit and the erythropoietic response to epoetin alfa (r2 = 0.06). Mean change in hematocrit after treatment with epoetin alfa in the quartile baseline hematocrit groups 34.2 to 41.4, 41.5 to 43.2, 43.3 to 44.9 and 45.0 to 48.0 hematocrit points was 3.71, 2.45, 3.86 and 1.02 hematocrit points, respectively. Of the surgical candidates 22 (9.1%) achieved an induction hematocrit of greater than 51%. Of the 283 men receiving epoetin alfa 21 (7.4%) also received an allogeneic transfusion. The transfusion rate did not correlate with induction hematocrit. The only adverse cardiovascular event was an uncomplicated postoperative pulmonary embolus. CONCLUSIONS: Our prospective study demonstrates that epoetin alfa given preoperatively in 2 doses of 600 IU/kg. is safe for significantly increasing hematocrit in men before radical retropubic prostatectomy. It is intuitive that the significant increase in hematocrit decreases the requirement for allogeneic blood transfusion. PMID- 10687988 TI - Dexamethasone does not significantly contribute to the response rate of docetaxel and estramustine in androgen independent prostate cancer. AB - PURPOSE: We evaluated the independent response rate of dexamethasone before docetaxel and estramustine administration as measured by changes in serum prostate specific antigen (PSA) in patients with androgen independent prostate cancer. MATERIALS AND METHODS: A total of 12 patients received 20 mg. dexamethasone orally every 6 hours for 3 doses repeated every 3 weeks before starting cytotoxic therapy with estramustine and docetaxel. After progression on dexamethasone 280 mg. estramustine orally 3 times daily on days 1 to 5 and 70 mg./m.2 docetaxel intravenously for 1 hour on day 2 were given. RESULTS: None of the patients initially treated with dexamethasone monotherapy (median 1 cycle, range 1 to 5) had a PSA decline of 50% or greater. Median PSA increase on monotherapy was 47% (range 0% to 22%). On estramustine and docetaxel therapy PSA decreased 50% or greater in 11 patients (92%, 95% confidence intervals [CI] 60 to 99) and 80% or greater in 7 (58%, 95% CI 29 to 84), and normalized in 5 (42%, 95% CI 16 to 71), with a median duration of response of 153 (range 42 to 371), 132 (range 84 to 287) and 84 (range 21 to 174) days, respectively. Median times to reach 50% and 80% decreases in baseline PSA were 21 (range 21 to 209) and 63 (range 21 to 138) days, respectively. In 9 patients (75%, 95% CI 43 to 93) PSA decreased at least 50% by week 9. Of 4 patients with bidimensionally measurable disease 3 had a partial response. Median time to progression was 263 days (range 91 to 378). CONCLUSIONS: Administration of 20. mg. dexamethasone orally every 6 hours for 3 doses every 3 weeks does not significantly contribute to the PSA response rate of estramustine and docetaxel. PMID- 10687989 TI - An implant releasing the gonadotropin hormone-releasing hormone agonist histrelin maintains medical castration for up to 30 months in metastatic prostate cancer. AB - PURPOSE: The administration of gonadotropin hormone-releasing hormone agonists is well established for treating metastatic prostate cancer. In an ongoing study we evaluated the effect of a long acting implant that releases the gonadotropin hormone-releasing hormone agonist histrelin ([ImBzl]D-His6,Pro9-Net) in 15 patients with disseminated prostate cancer. MATERIALS AND METHODS: The 2.6 cm. implant releasing 60 microg. histrelin daily is inserted subcutaneously into the upper arm using local anesthesia. Of the patients 8 received 1 and the remainder received 2 implants. Treatment with the antiandrogen flutamide or cyproterone acetate began 2 weeks before implant insertion and continued for up to 12 weeks. Testosterone, luteinizing hormone (LH) and prostate specific antigen were determined monthly, and a metastatic evaluation was performed every 6 months. RESULTS: LH and testosterone increased after flutamide administration and decreased after implant insertion. By day 28 LH and testosterone were completely suppressed. LH and testosterone decreased immediately after cyproterone acetate administration. Prostate specific antigen began to decrease during antiandrogen therapy and decreased further after implant insertion. One patient requested implant removal after 1 year for personal reasons and 1 died of an unrelated cause 18 months after insertion. Escape was demonstrated in 4 cases at 5, 10, 12 and 19 months, although LH and testosterone remained suppressed. Duration of treatment in the remaining 9 patients was between 21 and 30 months. LH and testosterone remained completely suppressed and prostate specific antigen levels were in the normal range. The clinical and biochemical response was identical in those who received 1 or 2 implants. At 12 months 8 patients were challenged at intermittent intervals for up to 24 months with a bolus of 100 microg. gonadotropin hormone-releasing hormone followed by 2 weeks of flutamide. The response was compared with that in untreated controls recently diagnosed with prostate cancer. Unlike controls there was complete LH suppression in the 8 challenged patients. CONCLUSIONS: A histrelin implant suppresses LH and testosterone in prostate cancer for up to 30 months. This finding represents a significant improvement over existing preparations, which must be administered at 1 to 3-month intervals. PMID- 10687990 TI - Radiotherapy for isolated serum prostate specific antigen elevation after prostatectomy for prostate cancer. AB - PURPOSE: Elevated serum prostate specific antigen (PSA) may be the initial and only indication of disease recurrence after prostatectomy for prostate cancer. External beam radiotherapy may be given in this setting in an attempt to eradicate the disease but therapeutic outcomes after this approach require further description. We describe the intermediate term outcome in a large group of patients treated with radiotherapy and identify pre-therapy factors associated with disease outcome. MATERIALS AND METHODS: We retrospectively studied a cohort of 166 consecutive patients treated with radiotherapy between July 1987 and May 1996. The Kaplan-Meier method was used to describe patient outcome for the overall study group, and statistical associations of pre-therapy variables with outcome were sought to identify predictive factors. RESULTS: At a median followup of 52 months 46% (95% confidence interval 38 to 55) of patients were expected to be free of biochemical relapse 5 years after radiotherapy. Multivariate analysis identified pathological classification (seminal vesicle invasion), tumor grade and preradiotherapy serum PSA as independent factors associated with biochemical relapse. Although in 1 of 6 patients a chronic complication was attributed to radiotherapy, it was often mild and self-limited in nature. CONCLUSIONS: In our current series approximately half of the patients treated with radiotherapy for an isolated elevation of serum PSA after prostatectomy were free of biochemical relapse at 5 years of followup. Radiotherapy may be given in this setting with modest long-term morbidity. PMID- 10687991 TI - Quality of life outcomes after brachytherapy for early stage prostate cancer. AB - PURPOSE: We compare general and disease specific health related quality of life in men undergoing brachytherapy for early stage prostate cancer to those undergoing radical prostatectomy and age matched healthy controls. MATERIALS AND METHODS: Cohorts consisted of 48 men treated with brachytherapy with and without pretreatment external beam radiation therapy (brachytherapy group), 74 who underwent radical prostatectomy (prostatectomy group) and age matched healthy controls from the literature. The RAND 36-item general health survey, University of California Los Angeles Prostate Cancer Index, American Urological Association symptom index, validated Cancer Interference with Life and Family Scales, and sociodemographic and co-morbidity questionnaires were completed 3 to 17 months after treatment. RESULTS: General health related quality of life did not differ greatly among the 3 groups. Urinary function (leakage) was worse in the brachytherapy group than in controls but better than in the prostatectomy group. Brachytherapy group patients had more irritative urinary symptoms and worse bowel function than controls. Sexual function and bother were worse in prostatectomy and brachytherapy groups than in healthy controls. Physical function, bodily pain, urinary function, and bother and American Urological Association symptom index scores improved with time after brachytherapy. Patients who underwent brachytherapy after external beam radiation performed worse in all general and disease specific health related quality of life domains compared to those who did not undergo pretreatment radiation therapy. CONCLUSIONS: At an average of 7.5 months after treatment the general health related quality of life of patients undergoing brachytherapy with and without pretreatment external beam radiation was similar to age matched controls, although urinary, bowel and sexual problems were reported. These problems appeared to improve during the first year after treatment. Much of the impairment in disease specific health related quality of life among patients undergoing brachytherapy may be attributed to pretreatment radiation. PMID- 10687993 TI - Experience with early catheter removal after radical retropubic prostatectomy. AB - PURPOSE: We tested the hypothesis that early catheter removal may be accomplished safely after radical prostatectomy. MATERIALS AND METHODS: Cystography on postoperative day 4 or 5 in 42 of 67 consecutive patients who underwent radical retropubic prostatectomy revealed no extravasation in 30 and the urethral catheter was removed (group 1). The control group included 25 patients who did not undergo cystography, and the catheter was removed 14 days postoperatively (group 2). RESULTS: Immediate and late continence was achieved in 14 (46.7%) and 25 (83.3%) cases in group 1, and in 8 (32%) and 22 (88%) cases in group 2, respectively (p>0.05). Catheterization was performed easily without any endoscopic or surgical procedure in 2 patients (6.7%) in group 1 who presented in urinary retention after catheter removal. Wound infection and pelvic abscess developed in 1 case (3.3%). There were no late complications. In group 2 urinary retention developed in 1 patient (4%), wound infection in 1 (4%) and hematuria in 1 (4%). Two patients (8%) had late vesical neck contracture at 4 and 10 months, respectively, which required urethrotomy in 1. In 1 patient (4%) a stricture in the anterior urethra was dilated. CONCLUSIONS: Our study shows that early catheter removal may be accomplished safely in most patients after radical retropubic prostatectomy, and was not associated with a higher complication rate. PMID- 10687992 TI - Multicenter patient self-reporting questionnaire on impotence, incontinence and stricture after radical prostatectomy. AB - PURPOSE: We determined the incidence of patient self-reported post-prostatectomy incontinence, impotence, bladder neck contracture and/or urethral stricture, sexual function satisfaction, quality of life and willingness to undergo treatment again in a large multicenter group of men who underwent radical prostatectomy. We also determined whether the morbidities of sexual function satisfaction, quality of life and bladder neck contracture and/or urethral stricture are predictable from demographic and postoperative prostate cancer factors. MATERIALS AND METHODS: A self-reporting questionnaire was completed and returned by 1,069 of 1,396 eligible patients (77%) who underwent radical prostatectomy between 1962 and 1997. Of the respondents 868 (85.7%) underwent surgery after 1990 and in all prostatectomy had been done a minimum of 6 months previously. Questionnaire results were independently analyzed by a third party for morbidity tabulation and the association of patient reported satisfaction. RESULTS: The patient self-reported incidence of any degree of post-prostatectomy incontinence, impotence and bladder neck contracture or urethral stricture was 65.6%, 88.4% and 20.5%, respectively. The incidence of incontinence requiring protection was 33% and only 2.8% of respondents had persistent bladder neck contracture or urethral stricture. Although incontinence and impotence significantly affected self-reported sexual function satisfaction, quality of life and willingness to undergo treatment again (p = 0.001), 77.5% of patients would elect surgery again. This finding remained true even after adjusting for demographic variables, and the time between surgery and the survey by multiple logistic regression. CONCLUSIONS: Although radical prostatectomy morbidity is common and affects self-reported overall quality of life, most patients would elect the same treatment again. Impotence and post-prostatectomy incontinence were significantly associated with sexual function satisfaction, quality of life and willingness to undergo treatment again. Bladder neck contracture and/or urethral stricture was associated with willingness to undergo treatment again after adjusting for demographic variables and time from surgery to the survey. PMID- 10687994 TI - The effect of hospital volume on mortality and resource use after radical prostatectomy. AB - PURPOSE: The value of radical prostatectomy for patients with prostate cancer depends on low morbidity and mortality. We assessed whether patient outcome is associated with how many of these procedures are performed at hospitals yearly. MATERIALS AND METHODS: Using the Nationwide Inpatient Sample, which is a stratified probability sample of American hospitals, we identified 66,693 men who underwent radical prostatectomy between 1989 and 1995. Cases were categorized into volume groups according to hospital annual rate of radical prostatectomies performed, including low-fewer than 25, medium-25 to 54 and high-greater than 54. We performed multivariate logistic regression to control for patient characteristics when assessing the associations of hospital volume, in-hospital mortality and resource use. RESULTS: Overall adjusted in-hospital mortality after radical prostatectomy was relatively low (0.25%). However, patients at low volume centers were 78% more likely to have in-hospital mortality than those at high volume centers (adjusted odds ratio 1.78, 95% confidence interval 1.7 to 2.6). Overall length of stay decreased at all hospitals between 1989 and 1995. However, average length of stay was longer and total hospital charges were higher at low than at high volume centers (7.3 versus 6.1 days, p<0.0001, and $15,600 versus $13,500, p<0.0001, respectively). CONCLUSIONS: Hospital volumes inversely related to in-hospital mortality, length of stay and total hospital charges after radical prostatectomy. Further study is necessary to examine the association of hospital volume with other important outcomes, including incontinence, impotence and long term patient survival after radical prostatectomy. PMID- 10687995 TI - Quality of life after prostatectomy. PMID- 10687996 TI - Urinary tract infection prophylaxis using Escherichia coli 83972 in spinal cord injured patients. AB - PURPOSE: Escherichia coli 83972 was previously shown to establish bladder colonization in select patient groups. We evaluate the safety and feasibility of using bacterial interference with E. coli 83972 to prevent urinary tract infection in spinal cord injured patients. MATERIALS AND METHODS: A total of 21 men and women with neurogenic bladder secondary to spinal cord injury underwent intravesical inoculation with E. coli 83972. Frequency of symptomatic urinary tract infection before and after colonization was compared. RESULTS: Successful long-term bladder colonization was achieved in 13 study participants. Mean duration of colonization was 12.3 months (range 2 to 40). Subjects had no symptoms of urinary tract infection while colonized with E. coli 83972 (0 infection per 18.4 patient-years). Successfully colonized subjects had experienced a mean of 3.1 symptomatic urinary tract infections per year (range 2 to 7) before colonization. Symptomatic infection also occurred in 4 subjects who were not successfully colonized with E. coli 83972 and in 7 others after spontaneous loss of colonization. Colonized subjects reported subjective improvement in quality of life with respect to urinary tract infection while colonized. CONCLUSIONS: E. coli 83972 may be safely used to establish long-term asymptomatic bladder colonization in spinal cord injured subjects. Preliminary findings suggest that colonization with E. coli 83972 may reduce the frequency of urinary tract infection in patients with neurogenic bladder secondary to spinal cord injury. PMID- 10687997 TI - A new technique for transurethral resection of superficial bladder tumor in 1 piece. AB - PURPOSE: We developed a new transurethral resection technique that not only removes the entire tumor, but also the surrounding mucosa and underlying stroma with superficial proper muscle in 1 piece to determine accurately the depth of invasion and infiltration pattern of superficial bladder cancer. MATERIALS AND METHODS: A short curved needle electrode is used to make a circular incision around and level incisions underneath the tumor, and for tumor retrieval. RESULTS: Tissue sections of the cut surface crossing the tumor center revealed accurate histology regarding the growth pattern and depth of invasion. No perforation, uncontrollable bleeding or other serious complications occurred. CONCLUSIONS: More accurate histological diagnosis regarding the growth pattern and depth of lamina propria invasion was possible with removal of the tumor and surrounding material in 1 piece compared to conventional transurethral resection. PMID- 10687998 TI - A simple technique to identify catheter balloon defects. PMID- 10687999 TI - Studies of the latency of pelvic floor contraction during peripheral nerve evaluation show that the muscle response is reflexly mediated. AB - PURPOSE: Whether neuromodulation using an implanted sacral nerve stimulator acts by its effects on pelvic afferent or efferent nerves remains to be determined. However, it has been observed that eliciting an "anal wink" is helpful in the optimal siting of the foraminal stimulating electrode. This observation has been interpreted as indicating that a direct effect on efferent pelvic innervation is an important functional component of the technique. We studied the latency of this motor response to determine whether it is consistent with neuromodulation working via a direct efferent mechanism. MATERIALS AND METHODS: We studied 9 women with urinary retention undergoing the first stage of a stimulator implant (peripheral nerve evaluation). Stimulation was applied to an electrode placed in the S3 foramen and the latency of the response of the striated anal sphincter, a contraction which gives rise to the "anal wink," was measured using a concentric needle electrode placed in the striated part of the anal sphincter. RESULTS: Mean latency of response was 98 milliseconds (range 52 to 140), which is approximately 10 times longer than would be expected from that resulting from direct motor nerve stimulation. CONCLUSIONS: Our results indicate that anal sphincter contraction observed during peripheral nerve evaluation is the result of an afferent mediated response. PMID- 10688000 TI - Surgical treatment for stress urinary incontinence associated with valsalva induced detrusor instability. AB - PURPOSE: Detrusor instability initiated by increased intra-abdominal pressure that results in incontinence has always been difficult to treat. This form of incontinence may be due to traction on the pelvic nerves when increased abdominal pressure is applied to already weakened pelvic supportive tissue. In most patients pharmacological attempts to correct this problem fail. We describe a pubovaginal sling designed to stabilize the urethrovesical junction during the Valsalva maneuver, which is our treatment of choice for such patients. MATERIALS AND METHODS: From 1994 to 1998 we treated 36 patients with a pubovaginal sling procedure for Valsalva induced detrusor instability diagnosed on preoperative urodynamics. The sling material was in situ vaginal wall in 20 cases, free swing vaginal wall in 6, rectus fascia in 4, cadaveric fascia in 3 and synthetic material in 3. Urodynamic evaluation was performed preoperatively in all patients. Followup of 6 months to 4 years involved subjective questions and objective examination. RESULTS: Cure was achieved in 33 of the 36 patients (92%), of whom leak point pressure was less than 50, 50 to 100 and greater than 100 cm. water in 9, 17 and 7, respectively. In the 3 failed cases leak point pressure was 50 to 100 cm water, including 2 in which cotton swab test results were less than 30 degrees. Urge incontinence resolved in 75% of the patients. CONCLUSIONS: The pubovaginal sling procedure may cure Valsalva induced detrusor instability. Leak point pressure does not determine which patients do well. Evaluation for hypermobility may help to predict the success or failure of a procedure by identifying those in whom Valsalva induced detrusor instability results from traction on the pelvic nerves. PMID- 10688001 TI - Report of the international consensus development conference on female sexual dysfunction: definitions and classifications. AB - PURPOSE: Female sexual dysfunction is highly prevalent but not well defined or understood. We evaluated and revised existing definitions and classifications of female sexual dysfunction. MATERIALS AND METHODS: An interdisciplinary consensus conference panel consisting of 19 experts in female sexual dysfunction selected from 5 countries was convened by the Sexual Function Health Council of the American Foundation for Urologic Disease. A modified Delphi method was used to develop consensus definitions and classifications, and build on the existing framework of the International Classification of Diseases-10 and DSM-IV: Diagnostic and Statistical Manual of Mental Disorders of the American Psychiatric Association, which were limited to consideration of psychiatric disorders. RESULTS: Classifications were expanded to include psychogenic and organic causes of desire, arousal, orgasm and sexual pain disorders. An essential element of the new diagnostic system is the "personal distress" criterion. In particular, new definitions of sexual arousal and hypoactive sexual desire disorders were developed, and a new category of noncoital sexual pain disorder was added. In addition, a new subtyping system for clinical diagnosis was devised. Guidelines for clinical end points and outcomes were proposed, and important research goals and priorities were identified. CONCLUSIONS: We recommend use of the new female sexual dysfunction diagnostic and classification system based on physiological as well as psychological pathophysiologies, and a personal distress criterion for most diagnostic categories. PMID- 10688002 TI - Ossifying cavernous hemangioma masquerading as adrenal tuberculosis. PMID- 10688003 TI - Unusual presentations of hydatid disease of the urinary tract. PMID- 10688004 TI - Orthotopic renal transplantation in a patient with a massive pelvic arteriovenous malformation. PMID- 10688005 TI - Adenocarcinoma in an Indiana pouch urinary diversion. PMID- 10688006 TI - Angiomyolipoma of the bladder wall. PMID- 10688007 TI - Squamous cell carcinoma of the urachus. PMID- 10688008 TI - Carcinoma of the female urethra responsive to moderate dose chemoradiotherapy. PMID- 10688009 TI - Priapism as a complication of high dose testosterone therapy in a man with hypogonadism. PMID- 10688010 TI - An unusual case of a metastatic lesion to the penis. PMID- 10688011 TI - Magnetic resonance imaging to diagnose segmental testicular infarction. PMID- 10688012 TI - Extragonadal germ cell tumor of mediastinum with high serum level of carcinoembryonic antigen and carbohydrate antigen 19-9. PMID- 10688013 TI - Basal cell carcinoma of the scrotum. PMID- 10688014 TI - Hemospermia and expressed tumor in the urethra: an unusual presentation of ductal carcinoma of the prostate. PMID- 10688015 TI - Media hype in the medical literature: what's a doctor to do? PMID- 10688016 TI - Sildenafil and the Internet. AB - PURPOSE: The Internet is changing the way medicine is being practiced and challenging our notions of the doctor-patient relationship. We analyze the development of online prescriptions and propose guidelines for the sale of sildenafil over the Internet. MATERIALS AND METHODS: Using MEDLINE, Medscape and Lexis-Nexis search engines we reviewed pertinent materials from January 1996 to July 1999 focusing on the keywords Viagra, prescription and Internet. The review included press releases, law review articles, case law, medical literature, pending litigation, proposed legislation, and federal and state statutes. RESULTS: Online prescriptions are an outgrowth of the mail order drug business. This development continues the historic innovations in communications and transportation that have enabled physicians to practice medicine over long distances while maintaining ties to hospitals and other specialists. While the sale of sildenafil over the Internet may be profitable and convenient, it raises a variety of legal, ethical and safety concerns. Many federal and state organizations have addressed the issue without establishing a clear standard. CONCLUSIONS: A clear distinction exists between online prescriptions and pharmacies. While it may be acceptable for sildenafil to be sold over the Internet given current technologies, it must be done within the confines of a traditional doctor-patient relationship. Online prescriptions must be limited to patients who live in states in which the prescribing physician is licensed. Failure to establish a doctor patient relationship in this context breeches ethical standards, and may give rise to potential civil and criminal liabilities. PMID- 10688017 TI - Re: unenhanced computerized axial tomography to detect retained calculi after percutaneous ultrasonic lithotripsy. PMID- 10688018 TI - Re: successful living related kidney transplantation despite renal angiomyolipoma in situ. PMID- 10688019 TI - Re: 13-year experience with percutaneous management of upper tract transitional cell carcinoma. PMID- 10688020 TI - Re: orthotopic urinary diversion with preservation of erectile and ejaculatory function in men requiring radical cystectomy for nonurothelial malignancy: a new technique. PMID- 10688021 TI - Re: orthotopic urinary diversion with preservation of erectile and ejaculatory function in men requiring radical cystectomy for nonurothelial malignancy: a new technique. PMID- 10688022 TI - Re: editorial: gender assignment and the pediatric urologist. PMID- 10688023 TI - Cystic testicular lesions in the pediatric population. AB - PURPOSE: We present the etiology, histological evaluation and management of all cystic lesions of the pediatric testis. MATERIALS AND METHODS: Illustrative cases from our experience are reported with a literature review of all possible diagnoses. RESULTS: Included in the differential diagnosis of cystic testis lesions in children are epidermoid cyst, dermoid cyst, prepubertal teratoma, juvenile granulosa cell tumor, cystic dysplasia of the rete testis, testicular cystic lymphangioma, simple cyst and cystic degeneration after torsion. Testis sparing surgery is feasible in many circumstances. CONCLUSIONS: Cystic lesions of the pediatric testis are rare but represent an interesting group of diagnoses. Patient age at presentation, examination features, tumor markers and sonographic appearance may assist in making a presumptive and occasionally definitive diagnosis preoperatively. Based on the likely diagnosis enucleation or partial orchiectomy may be considered when performed with frozen section histological assessment. A thorough understanding of potentially cystic testis lesions in children leads to the best management choices and often to preservation of a substantial portion of the affected testis. PMID- 10688024 TI - Comparison of laparoscopic versus open nephrectomy in the pediatric population. AB - PURPOSE: Laparoscopic renal surgery has become an accepted approach for benign disease in adults. We compare our experience with laparoscopic and open nephrectomy in a pediatric population. MATERIALS AND METHODS: A total of 10 pediatric patients underwent laparoscopic nephrectomy or nephroureterectomy and an additional 10 consecutive children underwent similar open procedures. All patients had benign disease and were treated at a single institution. Medical records were reviewed retrospectively for relevant clinical data. RESULTS: Planned surgery was completed in all cases. There were no conversions to open surgery in the laparoscopic group. Mean operative time was 175.6 versus 120.2 minutes (p = 0.01) and mean hospital stay was 22.5 versus 41.3 hours (p = 0.03) in the laparoscopic and open nephrectomy groups, respectively. Blood loss was not statistically different. Analgesic use was qualitatively less in the laparoscopic nephrectomy group. CONCLUSIONS: Laparoscopic nephrectomy and nephroureterectomy may be performed safely in children. While operative time was somewhat longer in our initial laparoscopic series, postoperative hospital stay was significantly shorter than for open surgery. Further experience with this technique is warranted. PMID- 10688025 TI - The modern endoscopic approach to ureterocele. AB - PURPOSE: During the last 20 years the surgical approach to ureterocele has evolved from major open surgery to minimally invasive endoscopic puncture. We believe that the endoscopic approach decreases the need for open surgical procedures. We identified specific factors that predict the need for repeat surgery. MATERIALS AND METHODS: We reviewed the charts of 60 new patients with ureterocele treated with primary endoscopic incision between 1991 and 1995. Followup ranged from 4 to 62 months (mean 20). Mode of presentation, ureterocele location, associated vesicoureteral reflux and association of the ureterocele with a duplex system were evaluated. Ureterocele wall thickness was assessed subjectively via radiographic and cystoscopic methods, and categorized as thin, intermediate and thick. RESULTS: All 9 patients with a single system ureterocele had an intravesical ureterocele. No patient had associated reflux nor did any require a secondary open procedure. In 3 cases new onset ipsilateral reflux into the ureterocele spontaneously resolved. Of the 51 patients with a duplex system and associated ureterocele 19 (37%) required a secondary open procedure. The ureterocele was intravesical and ectopic in 22 (43%) and 29 (57%) cases, respectively. Reflux was associated with the ureterocele in 27 patients (53%), and 12 (44%) required a secondary open procedure. A total of 11 patients underwent ureteral reimplantation of 15 refluxing renal units and only 2 renal units required ureteral tapering. Reflux is no longer present in 14 of the 15 renal units (93%). Patients with a thick walled ureterocele required repeat puncture more frequently than those with a nonthick ureterocele. CONCLUSIONS: With the use of modern endoscopic techniques children with intravesical and single system ureteroceles require secondary open surgery less frequently than those with ectopic and duplex system ureteroceles. The mode of presentation does not predict the need for a repeat open procedure. Thick walled ureteroceles require repeat endoscopic puncture more frequently than thin and intermediate walled ureteroceles. PMID- 10688026 TI - Bifid mesonephric duct presenting as a scrotal urinoma. PMID- 10688027 TI - Transureteroureterostomy in childhood and adolescence: long-term results in 69 cases. AB - PURPOSE: We analyzed a series of 69 transureteroureterostomies to evaluate long term results and specify current indications. MATERIALS AND METHODS: Between 1969 and 1998 transureteroureterostomy was performed in 32 females and 37 males with a mean age of 8.6 years. Surgery was done to avoid repeat or difficult ureteral reimplantation after multiple failed procedures in 22 cases and to create a continent ureteral conduit for intermittent catheterization in 23. Other indications included undiversion in 8 cases, ureterocystoplasty in 6, diversion in 4, the Kropp procedure in 3, massively dilated megaureter in 2 and ureteral necrosis in 1. Of the 69 patients 6 were lost to followup and 63 were followed at least 1 year (median 6). RESULTS: A total of 63 patients were regularly monitored by clinical observation and morphological investigation, including ultrasound, excretory urography and cystography. In 50 cases (79.4%) results were good with no upper urinary tract dilatation. All initially normal donor ureters remained normal. Of 51 initially dilated donor ureters 40 (78.4%) improved or returned to normal, while 20 of 27 initially dilated recipient ureters (74.1%) improved or returned to normal. Serious complications in 3 cases (4.3%) involved anastomotic leakage, ischemic stenosis of the common ureteral trunk and progressive deterioration of function in 1 kidney requiring nephrectomy 3 years postoperatively. Reoperation was successful in the former 2 cases. CONCLUSIONS: With careful attention to technique transureteroureterostomy represents a safe and reliable procedure with well-defined indications in pediatric urology. PMID- 10688028 TI - Y duplication of urethra with complete atresia of the orthotopic channel: 1-stage reconstruction. PMID- 10688029 TI - Hypospadias and urethral development. AB - PURPOSE: Hypospadias is a common congenital anomaly that may be treated with surgical reconstruction. In the majority of cases the etiology remains elusive. Although androgens are clearly critical for penile development, defects in androgen metabolism and/or the androgen receptor explain only a small subset of cases of hypospadias. Strategies are presented for understanding the etiology of hypospadias. MATERIALS AND METHODS: Current scientific reports on the etiology of hypospadias were reviewed, and the embryology and possible mechanisms of urethral and penile formation are presented. RESULTS: A new theory of glandular human urethral development via endodermal cellular differentiation is proposed to replace the classic explanation of ectodermal intrusion. CONCLUSIONS: Careful studies of penile and urethral development have led to a better understanding of genital embryology. Future areas of study, such as endocrine disrupters, mesenchymal-epithelial interactions and mechanisms of penile growth, are proposed to explain the etiology of hypospadias. PMID- 10688030 TI - Vanished testis: be aware of an abdominal testis. PMID- 10688031 TI - Laparoscopic diagnosis and clinical management of a solitary nonpalpable cryptorchid testicle in a postpubertal male. PMID- 10688032 TI - Ectopic and undescended testes: 2 variants of a single congenital anomaly? AB - PURPOSE: We compared pathological findings in ectopic and undescended testis to determine whether the pathological evidence supports the hypothesis that the 2 conditions are variants of the same congenital anomaly. MATERIALS AND METHODS: We identified 24 boys with ectopic testis not in the superficial inguinal pouch of Denis Browne. Seven boys were excluded from study due to unavailable clinical records for 3, contralateral undescended testis in 2 and inadequate biopsy specimens in 2. Pathological findings of ectopic testis in the remaining 17 patients were compared with those of age matched patients with unilateral undescended testis. Total germ cell count, testicular volume, patency of the processus vaginalis and epididymal abnormalities were compared. Data were analyzed using the Wilcoxon matched pairs signed rank and Fisher's exact tests. RESULTS: No difference was noted in total germ cell count (p = 0.33), testicular volume (p = 0.1475), processus vaginalis patency (p = 0.0854) or epididymal abnormalities (p = 1.00) in the 2 groups. Of the 24 boys (8%) with ectopic testis 2 also had a contralateral undescended testis. CONCLUSIONS: Similar pathological findings in ectopic and undescended testes as well as the association of ectopic testis with a contralateral undescended testis suggest that ectopic and undescended testes are variants of the same congenital anomaly. Thus, boys with ectopic testis may have an increased incidence of subfertility and testicular malignancy. This spectrum of abnormal testicular position, and its range of pathological conditions and complications may appropriately be called the undescended testis sequence. PMID- 10688033 TI - The predictive value of inguinal herniography for the diagnosis and treatment of cryptorchidism. AB - PURPOSE: We evaluated the role of a patent processus vaginalis for cryptorchidism as well as inguinal herniography as a predictor of the efficacy of human chorionic gonadotropin (HCG) treatment. MATERIALS AND METHODS: We studied 244 boys with unilateral and 66 with bilateral cryptorchidism. All patients underwent inguinal herniography and received HCG. Nonresponders to treatment subsequently underwent orchiopexy, when processus vaginalis status, testicular position and epididymal characteristics were assessed. RESULTS: HCG was effective for 139 of 281 testes (49.5%) with an obliterated and 0 of 95 with a patent processus vaginalis on herniography. We further evaluated herniography in accordance with orchiopexy findings of persistent unilateral and bilateral cryptorchidism in 206 boys (237 testes) after HCG. Herniography findings of processus vaginalis morphology revealed a close correlation with that reported by the surgeon after orchiopexy (p<0.000005). The incidence of a patent processus vaginalis increased as testicular position became more caudal. The processus vaginalis was obliterated in all cases of anorchia. The incidence of more severe epididymal anomalies decreased as the testicular position became more caudal. Epididymal abnormalities were more common when the processus vaginalis was patent. CONCLUSIONS: Pretreatment herniography assessment of processus vaginalis morphology is of prognostic value for predicting the efficacy of hormone treatment, the presence or absence of testes when they are nonpalpable and future fertility. PMID- 10688034 TI - Expression profile of an androgen regulated prostate specific homeobox gene NKX3.1 in primary prostate cancer. AB - PURPOSE: NKX3.1, a member of the family of homeobox genes, exhibits prostate tissue specific expression and appears to play a role in mouse prostate development. Rapid induction of NKX3.1 gene expression in response to androgens has also been described. On the basis of the established role of androgens in prostatic growth and differentiation and studies showing an association of aberrant homeobox gene expression with the neoplastic process, we hypothesize that alterations of NKX3.1 gene expression play a role in prostate tumorigenesis. MATERIALS AND METHODS: NKX3.1 expression was analyzed in matched, microdissected normal and tumor tissues from 52 primary prostate cancer specimens from radical prostatectomy by semiquantitative RT-PCR and in situ hybridization and correlated with the clinicopathologic features. NKX3.1 expression was quantified as differential expression between matched tumor and normal tissues and was grouped as overexpression in tumor tissue, reduced expression in tumor tissue and no change between tumor and normal tissues. Androgen regulation of NKX3.1 expression was also studied in LNCaP cells. Androgen receptor (AR) expression in prostate tumor and normal tissue was correlated with NKX3.1 expression. RESULTS: Comparison of NKX3.1 expression between normal and tumor tissues revealed overexpression in 31% tumor specimens (16 of 52), decreased expression in 21% tumor specimens (11 of 52) and no change in 48% specimens (25 of 52). When these expression patterns were stratified by organ confined and non-organ-confined tumor, a higher percentage of patients exhibited NKX3.1 overexpression in non organ confined tumor (40%) versus organ confined tumor (22%). Elevated NKX3.1 expression significantly correlated with tumor volume and serum prostate specific antigen (PSA) level in the NKX3.1 overexpression group (p<0.05). Metastatic prostate cancer cell lines did not exhibit mutations in the protein coding sequence of NKX3.1. Additionally, the NKX3.1 expression correlated with AR expression (p<0.01) in vivo in human prostate tissues. Comparison of PSA and NKX3.1 expression in response to androgen revealed a rapid androgen mediated induction of NKX3.1 expression in LNCaP cells. In situ hybridization analysis of representative specimens confirmed RT-PCR observations. CONCLUSIONS: These results suggest an association of NKX3.1 with a more aggressive phenotype of carcinoma of the prostate. Correlation of AR expression with NKX3.1 in human prostate tissues underscores the androgen regulation of NKX3.1 in the physiologic context of human prostate tissues. PMID- 10688035 TI - Characterization of ureteral dysfunction in an experimental model of congenital bladder outlet obstruction. AB - PURPOSE: Ureteral dysfunction is a significant sequela of congenital bladder outlet obstruction. However, the structural and functional alterations associated with ureteral dysfunction are not well defined. A model of fetal bladder obstruction in sheep was used to characterize the changes in ureteral smooth muscle, extracellular matrix (ECM) and functional properties in response to bladder outlet obstruction. MATERIALS AND METHODS: Partial bladder outlet obstruction was created in fetal sheep at gestational age 95 days via placement of a metal ring around the proximal urethra as well as ligation of the urachus. Ureters were harvested at 109 and 135 days (full term = 140 days) to determine the relative composition of smooth muscle, ECM and urothelium by morphometric analysis and to measure DNA and protein concentrations. Ureteral tissue from 135 day gestation obstructed and control sheep was harvested and immediately placed in Krebs solution. Smooth muscle strips (2-3 mm. x 7-8 mm.) were suspended in organ baths. The frequency and amplitude of spontaneous ureteral contractions was as well as the response to electric field stimulation (EFS) were determined. RESULTS: Bladder outlet obstruction caused a significant increase in ureteral weight, smooth muscle mass and total ECM at both 109 and 135 days gestation. Total ureteral DNA was greater in obstructed compared with sham ureters at 135 days gestation. Obstructed ureters demonstrated greater amplitude and frequency of spontaneous contractions as well as more pronounced response to EFS when compared to sham ureters. CONCLUSIONS: The fetal ureter responds to bladder obstruction with smooth muscle hyperplasia and hypertrophy which is associated with increased spontaneous activity and augmented response to EFS. ECM content is markedly increased indicating a shift in the balance of connective tissue synthesis and degradation. Congenital post-obstructive ureteral dysfunction therefore appears to be the result of dysregulated smooth muscle cell growth and altered ECM homeostasis producing abnormal ureteral contractility. PMID- 10688036 TI - Cell-cell adhesion molecules and signaling intermediates and their role in the invasive potential of prostate cancer cells. AB - PURPOSE: The highly variable natural history of prostate carcinoma may be reflected in heterogeneity of invasive potential between tumors. MATERIALS AND METHODS: We have examined two prostate cancer cell lines of low invasive potential (CAHPV10 and PZHPV7) and three cell lines of high invasive potential (DU-145, PC-3, LNCapFGC), to determine whether specific adhesion molecule profiles correlated with their invasive behavior. RESULTS: Using an in vitro invasion assay, we demonstrated that DU-145, LNCapFGC and PC-3 cells were highly invasive compared with CA-HPV-10 and PZ-HPV-7 cells. LNCapFGC cells expressed high levels of E-cadherin, alpha-, beta- and gamma-catenin, desmoglein, desmoplakin and GSK3beta using immunoblotting. This was, in general, comparable to immunohistochemical staining. PC-3 cells had no E-cadherin or alpha-catenin, but expressed a high level of the HGF/SF receptor c-Met. In contrast, DU-145 cells were found to express E-cadherin and low levels for all other protein molecules, except c-Met. The DU-145 cell line also lacked alpha-catenin expression. In CA-HPV-10 and PZ-HPV-7 cells, there was no detection of APC, PECAM 1, P-cadherin or Wnt-1. DU-145, LNCapFGC and PC-3 cells formed cell-cell aggregates, which were reduced by inclusion of anti-E-cadherin antibody and the motogen HGF/SF. CONCLUSION: These results show that prostate cancer cells exhibit a diverse expression of cell-cell adhesion molecules and their signaling intermediates. The expression of these adhesion molecules bears an important relationship with the invasive phenotype of these cells. PMID- 10688037 TI - Nuclear factor kappa B mediates lipopolysaccharide-induced inflammation in the urinary bladder. AB - PURPOSE: The proteins which constitute the final common pathway linking receptors on cell surfaces to the inflammatory cascade have recently been identified and cloned. Central to activation of this inflammatory cascade is translocation from cytosol to nucleus of the nuclear transcription factor known as nuclear factor kappa B (NF-kappaB). The purpose of this study was to determine whether NF-kappaB cascade plays a role in lipopolysaccharide (LPS)-induced inflammation of the mouse urinary bladder. MATERIALS AND METHODS: Bladder inflammation was induced in anesthetized mice by intravesical instillation of lipopolysaccharide (LPS) and quantified by morphometric analysis. The NK-1 receptors for substance P were quantified by flow cytometry. LPS-induced degradation of inhibitory IkappaB subunit was quantified by Western blotting analysis and translocation of NF kappaB protein from cytosol to the nucleus was determined by confocal microscopy and Western blotting analysis. In addition, we determine the effect of lactacystin, a proteosome inhibitor, on LPS-induced IkappaB degradation and NF kappaB translocation, NK-1 receptor fluorescence intensity, and bladder inflammation. RESULTS: LPS instillation into the mouse bladder resulted in time dependent loss of the inhibitory IkappaB subunit of the NF-kappaB protein complex. IkappaB cleavage was followed by translocation of NF-kappaB from the cytosol to the nucleus. This was associated with increased expression of an NF kappaB dependent inflammatory component, the NK-1 receptor. Pretreatment of mouse bladders with the NF-kappaB inhibitor, lactacystin, prevented cleavage of IkappaB in a dose-dependent manner. Lactacystin prevented increases in the NF-kappaB dependent inflammatory cascade components such as the NK-1 receptor. At the whole tissue level, the marked inflammatory infiltrate and mucosal breakdown associated with LPS administration was completely abolished by lactacystin. CONCLUSION: NF kappaB mediates many features of urinary bladder inflammation induced by LPS. The NF-kappaB cascade is an important target for anti-inflammatory management of cystitis. PMID- 10688038 TI - The effect of mycophenolate mofetil and polyphenolic bioflavonoids on renal ischemia reperfusion injury and repair. AB - PURPOSE: Chronic renal allograft nephropathy is associated with both immune and ischemic injury which may act synergistically to promote an inflammatory response. The immunosuppressant mycophenolic acid and the polyphenolic agents curcumin and quercetin possess properties that might ameliorate such injury. We studied the effects of these agents in models of ischemic renal injury and skin allograft rejection. MATERIALS AND METHODS: Ischemic acute tubular necrosis was produced in rats by renal pedicle occlusion with contralateral nephrectomy. Animals were treated with mycophenolic acid, quercetin and curcumin, or a combination of agents. Animals were killed on days 2 and 7 after operation and tissue samples were collected. Renal tubular apoptosis and cellular proliferation were assessed by immunohistochemistry. Expression of the cytokines RANTES and AIF were evaluated by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Treatment with the polyphenolic compounds alone or in combination with mycophenolic acid was associated with less tubular damage, attenuation of renal inflammation, and prolongation of skin graft survival. A combination of agents decreased serum creatinine on day 2 from 4.5 to 0.9 mg./dl. (p< or =0.01) and at day 7 from 3.8 to 0.6 mg./dl. (p< or =0.01). Treatment with the polyphenolic compounds inhibited apoptosis at day 2. By RT-PCR, RANTES and AIF were detected at high levels on days 2 and 7. Treatment with these agents alone or in combination strongly attenuated this increased expression. CONCLUSIONS: The combination of mycophenolic acid with curcumin and quercetin reduces renal injury and facilitates repair. These agents may have a role in therapeutic regimens that are both immunosuppressive and renoprotective. PMID- 10688039 TI - Hydro-jet assisted laparoscopic partial nephrectomy: initial experience in a porcine model. AB - PURPOSE: Hemostasis represents a challenge when performing laparoscopic partial nephrectomy. Hydro-Jet cutting is an advanced technology that has been used to create an ultra-coherent water force that functions like a sharp knife. In the surgical field, it has mainly been used for liver surgery and initial clinical experience with laparoscopic cholecystectomies has been favorable. This technique allowed selective parenchymal cutting with preservation of vessels and bile ducts. We describe a novel Hydro-Jet assisted dissection technique for laparoscopic partial nephrectomy in a porcine model. MATERIALS AND METHODS: Ten partial nephrectomies were performed in 5 pigs using a Muritz 1000 (Euromed Medizintechnik, A. Pein, Schwerin, Germany) Hydro-Jet generator. A thin stream of ultra coherent fluid is forced at a high velocity through a small nozzle. A modified probe allows both blunt dissection concomitantly with high-pressure water application. Coagulation can be applied via a bipolar thermoapplicator as needed. RESULTS: Laparoscopic partial nephrectomy was successful in all animals. Water-jet cutting through the parenchyma was virtually bloodless and preserved the vasculature and the collecting system. The vessels were then ligated or coagulated under direct vision. The continuous water flow established a bloodless operating field and a clear view for the surgeon. The mean dissection time and warm ischemia time were 45+/-9 and 17+/-3 minutes, respectively. CONCLUSIONS: This preliminary study supports the suitability of this technique for laparoscopic partial nephrectomy to improve hemostasis. The improved anatomical dissection and hemostasis may further decrease morbidity and operative time. Further studies are underway to compare this technique with laser coagulation for laparoscopic partial nephrectomy. PMID- 10688040 TI - Recruitment, distribution and phenotypes of mast cells in interstitial cystitis. AB - PURPOSE: Interstitial cystitis (IC) is a chronic disabling condition of unknown etiology. One of its major characteristics is an increase in mast cells (MC) showing signs of activation. It has been suggested that the proteinase content defines two MC types: MC(TC), containing chymase and tryptase, and MC(T), which contains tryptase but lacks chymase. Here, we investigated the MC distribution and the MC proteinase expression in IC together with the tissue expression of the major MC growth factors, stem cell factor (SCF) and interleukin-6 (IL-6). MATERIALS AND METHODS: MC were enumerated in bladder specimens from patients with classic IC, nonulcer IC and controls. MC were visualized in terms of metachromasia, reflecting glycosaminoglycan content, and immunohistochemically, visualizing tryptase, chymase and IL-6 as well as the surface markers CD117 and SCF. RESULTS: Classic IC displayed a 6 to 10-fold increase of MC identified by proteinase content while in nonulcer IC there were twice as many MC as in controls. In contrast to nonulcer IC and controls, classic IC displayed an abundance of epithelial MC. Fewer CD117+ than proteinase+ MC were detected in IC but not in controls. Classic IC coexpressed SCF and IL-6 in the epithelium and displayed numerous SCF and IL-6+ cells in the mucosa and detrusor muscle, many of which were MC. CONCLUSIONS: Redistribution of MC into the epithelium and a high bladder wall MC density distinguish classic IC from nonulcer IC. Our findings suggest an SCF/IL-6-driven MC response in IC. They also indicate a downregulation of the SCF receptor in IC. PMID- 10688041 TI - Immunobiologic, cytogenetic and drug response features of a newly established cell line (SCRC-1) from renal small cell carcinoma. AB - PURPOSE: We describe the establishment and preliminary characterization of a cell line designated SCRC-1, which was derived from a primary renal small cell carcinoma. MATERIALS AND METHODS: Continuous cultures of a primary stage IVa renal small cell carcinoma and a xenograft in nude mice derived therefrom were characterized by immunohistology, electron microscopy, immunofluorescence/flow cytometry, cytogenetic analysis, and an in vitro drug resistance assay. RESULTS: SCRC-1 cells were reactive with antibodies to NSE, chromogranin-A, bombesin, Bcl 2, CD44s, CD44v6, CD44v7 to 8, vimentin and S100 protein (predominantly beta subunit), and were unreactive with antibodies to EMA, CD54, EGFR(R1), URO-5, URO 7, URO-8 and URO-10. A similar immunoprofile was also found in both the primary tumor and the xenograft. Cytogenetic analysis revealed the following common clonal aberrations in all 50 metaphases analyzed: 45, XX, t (X;10;18) (p11;p11;q11), -der(18)t(X;10;18), indicating the clonal nature of this neoplasm. SCRC-1 cells showed low drug resistance to cyclophosphamide, doxorubicin, gemcitabine and fluorouracil, intermediate resistance to carmustine and mitomycin C, and extreme resistance to cisplatin. CONCLUSION: We have documented the initial characterization of SCRC-1, which may be the first cell line reported to be derived from a primary small cell carcinoma of the kidney. This cell line can be used for further studies uncovering the biology and histogenesis of this rare cancer and delineating differences among small cell carcinomas of the kidney and other histological types. PMID- 10688042 TI - Efficacy of microtubule-active drugs followed by ketoconazole in human metastatic prostate cancer cell lines. AB - PURPOSE: Once a relapse occurs following primary endocrine treatment, metastatic prostate cancer is one of the most therapy-resistant human neoplasms. Ketoconazole is used for complete androgen deprivation, and recent data suggest it has direct activity against prostate cancer cells. MATERIALS AND METHODS: LNCaP, DU145, and PC3 cells, human prostate cancer cell lines, and HL60, a human leukemia cell line, were lysed and soluble proteins were harvested. Cells were plated in 96-well flat bottom plates and then exposed to the pharmacological agents, ketoconazole, vinblastine and paclitaxel. DNA synthesis was monitored by 3H-thymidine incorporation. RESULTS: We demonstrate that ketoconazole exerts a cytostatic effect on a panel of human prostate cancer cell lines, with IC50 of 4 to 5 microg./ml., 12 microg./ml., and 25 microg./ml. for LNCaP, PC3/PC3M, and DU145 cells, respectively. On the other hand, using microtubule-active drugs, vinblastine and paclitaxel, we found that PC3M and PC3 cells were more resistant than either DU145 or LNCaP cells. This resistance was associated with a lesser degree of Raf-1 and Bcl-2 phosphorylation following exposure to microtubule active drugs. Combinations of microtubule-active drugs with ketoconazole were a beneficial treatment in DU145 cancer cells. Furthermore, ketoconazole blocked recovery of all the prostate cancer cell lines following 24 hours-pulse treatment with vinblastine. CONCLUSION: Pulse-administration of vinblastine followed by continuous administration of ketoconazole warrants investigation in the treatment of hormone-independent metastatic prostate cancer. PMID- 10688043 TI - Mitogenic signaling in androgen sensitive and insensitive prostate cancer cell lines. AB - PURPOSE: To investigate the role of a specific mitogen activated protein kinase, extracellular signal-regulated kinase (ERK), in regulating cell proliferation induced by three potentially important prostate cancer mitogens that signal via different classes of receptors. MATERIALS AND METHODS: Androgen sensitive (LNCaP) and insensitive (PC-3) prostate cancer cell lines were used in these studies. Epidermal growth factor (EGF), lysophosphatidic acid (LPA), and dihydrotestosterone (DHT) were the mitogenic stimulants and AG1478, a receptor tyrosine kinase inhibitor, and PD98059, an inhibitor of MEK, were the chemical inhibitors used in this study. Cell proliferation was measured using the WST-1 assay and ERK expression and activation was determined by immunoblotting for phospho- and total ERK. RESULTS: In androgen-sensitive LNCaP cells, epidermal growth factor (EGF) and dihydrotestosterone (DHT) both enhanced cell proliferation. EGF-stimulation dramatically increased ERK phosphorylation while DHT did not. In the androgen-insensitive cell line, PC-3, EGF- and LPA-induced ERK phosphorylation and cell proliferation. Inhibition of EGF- and LPA- induced ERK activation with the EGF receptor inhibitor, AG1478, or the MEK inhibitor, PD98059, attenuated their proliferative effects. Neither inhibitor had an effect on DHT stimulated cell proliferation. CONCLUSIONS: These data demonstrate heterogeneity of mitogenic signaling in prostate cancer cells, and support the hypothesis that androgens and growth factors utilize divergent signaling pathways in prostate cancer to induce proliferation. PMID- 10688044 TI - The growth inhibitory effect of p21 adenovirus on human bladder cancer cells. AB - PURPOSE: To evaluate whether p21 (WAF-1/CIP1) should be considered a potential candidate for human bladder cancer gene therapy, we determined: (1) the basal level of p21 expression in bladder cancer cell lines, (2) the response of bladder cancer cells to increased p21 expression following p21 adenovirus infection, and (3) the mechanism of growth inhibition produced by p21 overexpression. MATERIALS AND METHODS: Five established human bladder cancer cell lines and one primary culture derived from an invasive transitional cell carcinoma were used in this study. To examine the effect of p21 protein on the growth of human bladder cancer cells, a recombinant adenovirus vector system containing p21 cDNA, under the control of cytomegalovirus promoter, was constructed. A control virus containing p21 in an antisense orientation was used to eliminate potential artifacts caused by viral toxicity. RESULTS: Human bladder cancer cell lines exhibit variable endogenous p21 levels which correlate with the in vitro growth status. Significant, but highly variable increases in the steady-state level of p21 were detected in p21 adenovirus infected cells. Human bladder cancer cell lines responded heterogeneously to p21 adenovirus infection. Growth of the WH cell line was substantially inhibited in a dose and time-course dependent fashion. The mechanism of p21 growth inhibition was found to be due to G0/G1 arrest and not the induction of apoptosis. In contrast, p21 adenovirus failed to inhibit the growth of T24 bladder cancer cells because T24 cells were resistant to viral infection. The 253J bladder cancer cells exhibited marked sensitivity to adenovirus; substantial growth inhibition was seen with both sense and antisense p21 very early in the time course of infection. CONCLUSIONS: We found significant variation in the basal level of p21 protein expression in several human bladder cancer cell lines. Increased p21 expression as a result of adenoviral infection may be a potent growth suppressor in some human bladder cancer because it elicits cell cycle arrest in G0/G1 stage, but not the induction of apoptosis. Bladder cancer cells exhibit a wide spectrum of sensitivity to adenoviral infection that may be caused by the presence of viral receptor heterogeneity. This wide spectrum of sensitivity has significant basic scientific and clinical implications and warrants further study. PMID- 10688045 TI - Detection of loss of heterozygosity in the p53 tumor-suppressor gene with PCR in the urine of patients with bladder cancer. AB - PURPOSE: Detection of loss of heterozygosity (LOH) has been described in various carcinomas on the basis of meticulous molecular techniques. Because of lack of simple and rapid techniques, LOH has not achieved common use in routine tumor diagnosis. A recently found variable number of tandem repeats (VNTR) segment in intron 1 of the p53 gene was described as highly polymorphic and therefore useful in detecting LOH. We used a rapid technique for detection of LOH in the p53 gene of patients with transitional cell carcinoma (TCC) of the bladder. The technique was based on the polymerase chain reaction (PCR) and agarose gel electrophoresis as described for other carcinomas previously. We evaluated whether TCC screening and surveillance could be performed detecting LOH in the urinary sediment. MATERIALS AND METHODS: We investigated 29 patients with TCC of the bladder (pTa 12 patients; pT1 10 patients; pT2 - pT4 seven patients; grade 1 one patient; grade 2 19 patients; grade 3 nine patients). DNA was prepared by standard methods from white blood cells, tumor tissue, normal bladder mucosa, and urinary sediments. The amplification of the VNTR region was performed with PCR. PCR products were run in parallel lanes on 4.5% agarose gels. RESULTS: Of the 29 patients, 23 (79.3%) were found to have two different alleles ("informative cases") for the VNTR region. Of the 23 informative cases LOH was detected in the tumor tissue of 10 patients (43.5%). Referring to the total population 10 of 29 patients (34.4%) revealed LOH. In all patients with LOH in the tumor, LOH was also detected in the urinary sediment. LOH was not detected in the histologically benign bladder mucosa. CONCLUSION: We present a simple and rapid technique based on PCR and agarose gel electrophoresis for the detection of LOH in tumor and urinary sediment of patients with TCC of the bladder. The ability to detect LOH not only in tumor tissue but also in urinary sediment offers an attractive approach for noninvasive diagnosis and surveillance of bladder cancer patients. PMID- 10688046 TI - Muscle weakness, hyperactivity, and impairment in fear conditioning in tau deficient mice. AB - Tau, one of the major neuronal microtubule-associated proteins (MAPs), is important for neuronal cell morphogenesis and axonal maintenance. Tau is also known to be a component of the paired helical filaments (PHFs) in Alzheimer's disease patients. Recently, mutations in the tau gene were found in a hereditary neurodegenerative disease called frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) which exhibits various neurological and neuropathological characteristics including PHF-like intracellular tau deposit formation. Currently, the phenotype of the disease is thought to be due to: (1) the toxicity of mutant tau molecules and and/or; (2) the loss of function of normal tau molecules in patients' brains. To test the latter hypothesis, we performed behavioral and neurological tests on tau-deficient mice. Tau-deficient mice showed muscle weakness in the wire-hanging test, hyperactivity in a novel environment, and impairment in the contextual fear conditioning. They also had a tendency to fall more easily in the rod-walking test. These phenotypes parallel some signs and symptoms of FTDP-17 patients. Our results show that the loss of tau protein may itself lead to some of the neurological characteristics observed in FTDP-17 patients. PMID- 10688047 TI - The deletion polymorphism and Val1000Ile in alpha-2-macroglobulin and Alzheimer disease in Caribbean Hispanics. AB - The association between polymorphisms in the alpha-2-macroglobulin (a2m) gene and Alzheimer's disease remains in doubt because of conflicting results in independent case-control and family studies. We examined the association between Alzheimer's disease and alpha2m polymorphisms in Caribbean Hispanic families. The odds of having the alpha2m deletion/insertion polymorphism was increased 3-fold for family members with Alzheimer's disease compared to healthy family members, rising to 5-fold after adjusting for APOE-epsilon4. In contrast, there was no relationship between the alpha2m Val1000Ile polymorphism and Alzheimer's disease in these families. The inconsistencies in studies cited above and the modest association between alpha2m and Alzheimer's disease found in the Caribbean Hispanic families reported here, suggest that the overall effect of this gene on susceptibility is small and may be limited to certain populations or families. PMID- 10688048 TI - The response of growth hormone and prolactin of rats to hypoxia. AB - Effects of acute and chronic hypoxia on growth hormone (GH) and prolactin (PRL) of male rats were studied in a simulated hypobaric chambers at altitudes of 5 km (10.8% O2) and 7 km (7.2% O2), respectively. Acute hypoxia caused decreased plasma GH and increased pituitary GH content; both pituitary and plasma PRL contents at 2 h were decreased and plasma PRL level increased at 24 h. Prolonged exposure of hypoxia (5 km) to 25 days, both pituitary and plasma GH were obviously lower than control and pituitary PRL levels were decreased but plasma PRL increased markedly. The data presented suggest that long-term of hypoxia (10.8% O2) significantly suppresses body growth of rats and inhibits GH release and/or biosynthesis, which may in part correlate with decreased body weight gain; high circulating PRL concentration may be of significance in physiological adaptation to chronic hypoxia. PMID- 10688049 TI - Preoperative open field behavior predicts levels of neuropathic pain-related behavior in mice. AB - Exploratory open field (OF) activity was assessed in seven different mouse strains and selection lines. We counted the number of beam interruptions made by three cagemate mice at a time. This assay tests reactivity to aversive stimuli, anxiety and emotionality. One hindlimb was then totally denervated by transecting the sciatic and saphenous nerves on one side, and autotomy, a behavior thought to be related to neuropathic pain, was quantified over 35 days. We report that OF activity and autotomy are highly variable across different strains/lines. These results reaffirm the genetic control of these behaviors. We also found that these behaviors are inversely and significantly correlated. We suggest that common genetically-determined neural mechanisms may underlie anxiety, emotionality and neuropathic pain in mice. PMID- 10688050 TI - Effects of ascorbate in microdialysis perfusion medium on the extracellular basal concentration of glutamate in rat's striatum. AB - There are many evidences suggest that ascorbate in the extracellular space can affect glutamate concentration in the rat's brain. In this report, we studied how ascorbate in microdialysis perfusion medium affected glutamate level at the striatum in freely-moving rats. Three perfusion mediums were used: 0, 250, and 400 microM of ascorbate in perfusion medium. The extracellular basal concentrations of glutamate were determined to be 1.29+/-0.52 microM for the no ascorbate group, 0.86+/-0.35 microM for the low ascorbate group and 4.76+/-1.48 microM for the high ascorbate group. By using 400 microM of ascorbate in a perfusion medium, we found that the extracellular basal concentration of glutamate significantly increased and its in vivo recovery significantly decreased. This indicated that ascorbate concentration in a perfusion medium was important and must be carefully considered while using microdialysis technique to monitor glutamate concentration in vivo. PMID- 10688051 TI - The difference between electroacupuncture only and electroacupuncture with manipulation on analgesia in rats. AB - Plain acupuncture uses manipulation (rotation or varying the depth of insertion of the needle) to increase its effect. However, in commonly used electroacupunture (EA), variable manipulations have not been used. This study was performed to investigate the possibility of an increase in analgesic effect by adding manipulation to EA. The pain index used was the Tail-Flick latency (TFL) of the rat, which was lightly anesthetized with thiopental sodium (intraperitoneally). Four types of manipulation were used. Rotation and varying the depth of the needle (RN and VN) was employed using two different types of manipulation during each 20 min stimulation of EA. Each manipulation persisted for 1 min out of every 5 min (long - duration and long - interval: LDLI) or 12 s every 1 min (short - duration and short interval: SDSI). EA produced an increase in TFL; peak value was 49.7+/-12.2% of the pre - EA and occurred immediately after cessation of 20 min of EA stimulation. Performing RN or VN combined with EA also increased TFL more than just EA and a greater peak increase in TFL was observed with a SDSI - RN and SDSI - VN as compared to a LDLI - RN and LDLI - VN (77.5+/-13.8, 79.2+/-19.8 and 67.3+/-14.0%, 65.6+/-23.7% of the pre - EA, respectively). These results indicate that manipulation combined with EA produces a more potent antinociception than when only EA is applied. PMID- 10688052 TI - Different generators in human temporal-parasylvian cortex account for subdural laser-evoked potentials, auditory-evoked potentials, and event-related potentials. AB - In order to localize cortical areas mediating pain we now report subdural cortical potentials evoked by auditory stimulation (auditory-evoked potentials - AEPs) and by cutaneous stimulation with a laser (laser-evoked potentials - LEPs). Stimulation with the laser evokes a pure pain sensation by selective activation of nociceptors. LEPs were maximal over the inferior aspect of the central sulcus and had the same polarity on either side of the sylvian fissure. AEPs were maximal posterior to the LEP maximum and had opposite polarity on opposite sides of the sylvian fissure, consistent with the location of a known generator in the temporal operculum. Auditory P3 (event-related) potentials were maximal over the temporal base. These findings demonstrate that the LEP generator is not in secondary somatosensory cortex on the parietal operculum and is different from the P3 generator. PMID- 10688053 TI - Magnitude effects of galvanic vestibular stimulation on the trajectory of human gait. AB - This study examines the contribution of the vestibular system during different magnitudes of galvanic vestibular stimulation (GVS) during human walking. Anodal threshold levels of GVS were determined for right and left sides for each subject. Seven conditions were tested (no stimulation, left and right anode stimulation) at one, two and three times threshold. GVS was delivered to the mastoid processes at first heel contact and continued for the duration of the trial. All subjects responded by deviating towards the anode while walking. In addition, the magnitude of deviation increased as the stimulus intensity increased. Our results demonstrate that the vestibular system is sensitive to GVS intensity changes and responds by altering the magnitude of the response accordingly. These data provide a strong argument in support of a significant role for vestibular information during dynamic tasks. PMID- 10688054 TI - Bilateral tetrodotoxin blockade of the rat vestibular nuclei substitutes the natural unconditioned stimulus in taste aversion learning. AB - The aversive effects of bilateral transient blockade of the lateral vestibular nucleus caused by tetrodotoxin microinjections were tested using conditioned taste aversion in the first experiment. Male Wistar rats received tetrodotoxin injections (10 ng) after drinking a coffee solution (0.5%), either in the lateral vestibular nucleus (LVN), the parabrachial nucleus or the dopaminergic area A8. Two days later they drank a cider vinegar solution (3%) not followed by injections. In a later choice test, only the group receiving the injection in the lateral vestibular nucleus displayed a coffee aversion. In a second experiment the role of the peripheral vestibular symptoms induced by LVN inactivation on substituting the aversive stimulus was explored in the same behavioral task. Rats anesthetized (Pentobarbital, 25 mg/kg) before tetrodoxin LVN blockade, that did not show peripheral symptoms, did not develop learned aversions. The coffee preference ratios did not differ to those animals receiving only anesthesia or those that remained undisturbed. These results showed that the bilateral blockade of the vestibular nuclei may induce peripheral vestibular symptoms that that may substitute the aversive stimulus in taste aversion learning. PMID- 10688055 TI - Block by extracellular Mg2+ of single human purinergic P2X4 receptor channels expressed in human embryonic kidney cells. AB - Single channel properties of human P2X4 receptors expressed in human embryonic kidney cells have been investigated by outside-out mode patch clamp recordings. P2X4 channel activity was characterized by very fast kinetics. The current voltage relationship was strongly non-linear at potentials <-100 mV. A slope conductance of approximately 9 pS was estimated at the approximately linear part of the current-voltage relation (>-100 mV). External Mg2+ reversibly decreased the amplitude of ATP-evoked single channel currents in a concentration-dependent manner but independent of the membrane potential. Additionally, extracellular Mg2+ shortened the mean open time whereas the mean closed time was not affected. Thus, Mg2+ ions are proposed to inhibit the function of human P2X4 receptors by means of an open-channel block with a Mg2+ binding site at the exterior surface of the pore. PMID- 10688056 TI - Amino acid neurotransmitter metabolism in neurones and glia following kainate injection in rats. AB - Limbic seizure was induced in rats by intraperitoneal injection of the glutamate receptor agonist kainic acid. After 14 days [1-13C]glucose and [1,2-13C]acetate were injected subcutaneously and the rats killed 15 min later. Analysis of brain extracts was performed using 13C-magnetic resonance spectroscopy and high performance liquid chromatography. No significant differences between the two groups of rats were found for label concentration in blood or total metabolite tissue levels. Only astrocytes are able to utilize acetate as a substrate, whereas glucose is thought to be metabolized predominantly in the neuronal tricarboxylic acid cycle. Thus information about neuronal and astrocytic metabolism could be obtained in the same animal. A significant increase in label derived from [1-13C]glucose was observed in metabolites such as glutamate, gamma aminobutyric acid, aspartate, and succinate (all of which are mainly labelled in neurones). The increased labelling of glutamine in epileptic rats might be due to transfer of labelled glutamate from neurones to astrocytes. Astrocytic metabolism of acetate and transfer of glutamine to neurones were not affected. The results suggest that increased neuronal activity 2 weeks following epileptic seizures produces increased amino acid turnover in neurones. Changes in astrocytic metabolism were not detected. PMID- 10688057 TI - Gabapentin, an antiepileptic drug, improves memory storage in mice. AB - Male CF-1 mice were tested 48 h after training in a one-trial step-through inhibitory avoidance task. Immediately post-training i.p. injections of the antiepileptic drug gabapentin (1-aminomethyl cyclohexaneacetic acid) (GBP; 5, 10, 50, and 100 mg/kg) induced a dose-dependent enhancement of retention performance. Gabapentin did not affect response latencies in mice not given the footshock on the training trial, indicating that the actions of GBP on retention were not due to non-specific proactive effects on response latencies. The effects of GBP (10 mg/kg) were time-dependent, and the administration of GBP (10 mg/kg) 30 min before training also enhanced retention performance. However, the administration of GBP (10 mg/kg) 30 min prior to the retention test did not modify retention latencies of mice that had received either saline or GBP (10 mg/kg) immediately after training. Altogether, the results suggest that GBP influences retention by modulating time-dependent processes involved in memory storage, although the mechanism(s) of this action remain to be established. PMID- 10688058 TI - Decreased ability of rat temporal hippocampal CA1 region to produce long-term potentiation. AB - Tetanic stimulation of Schaffer collaterals in the CA1 region of transverse slices, taken from the septal (dorsal) part of young rat hippocampus, produced N Methyl-D-aspartate-dependent long-term potentiation (LTP) of the rising slope of excitatory postsynaptic potential (mean 38%). Under identical conditions of stimulation (100 Hz, 1 s) slices taken from the temporal (ventral) third of hippocampus presented a substantially reduced ability for LTP (mean 5%). The defect appeared to lie with the induction rather than the maintenance phase of LTP. These results suggest that a significant functional differentiation at the local synaptic plasticity level occurs between the two poles of hippocampus, which together with the substantial differences in their extrinsic connections, may help explain the reported differential participation of neurons in these parts of hippocampus during animal memory tests. PMID- 10688059 TI - The effects of low frequency and two-pulse stimulation protocols on synaptic transmission in the CA1-subiculum pathway in the anaesthetized rat. AB - The downregulation of synaptic efficacy is referred to as long-term depression (LTD). Recent work has shown that a two-pulse stimulation (TPS) protocol is successful at inducing LTD in vivo in area CA1 of the hippocampus. Here, we examine the ability of two TPS protocols and two low-frequency stimulation (LFS) protocols to induce LTD in the projection from hippocampal area CA1 to the subiculum in the anaesthetized rat. We find no evidence of LTD induction with TPS or LFS protocols. Instead, with three of the protocols (both TPS protocols and 1 Hz LFS), a late-developing potentiation is observed. PMID- 10688060 TI - Continuous exposure to dim illumination uncouples temporal patterns of sleep, body temperature, locomotion and drinking behavior in the rat. AB - Dissociable circadian rhythms of sleep and body temperature in primates are thought to be regulated by independent oscillators whereas the uncoupling of circadian rhythms has not been well described in other mammals. Therefore, we made simultaneous recordings of non-rapid-eye-movement-sleep (NREMS), rapid-eye movement-sleep (REMS), brain temperature, intraperitoneal temperature, locomotion and drinking activity under light-dark (LD) and continuous dim illumination (dim LL) and analyzed their interrelations. The rhythmic patterns of body temperature, locomotion and drinking were modified on the 12th circadian day of dim LL, while the mean body temperature as well as mean occurrence of drinking and locomotor activities did not change significantly. In contrast, dim LL exposure significantly increased the total time spent in NREMS during the resting phase of dim LL and increased REMS episodes during the active phase of dim LL. The diverse effects of dim LL exposure on the recorded phenomena suggest that temporal patterns of sleep were the most sensitive to perturbations of lighting and that differential oscillatory mechanisms may regulate sleep and other circadian rhythms in the rat. PMID- 10688061 TI - Food conditioning is impaired in cats deprived of the taste of food in early life. AB - Cats were fed by stomach tube during the first 75 days of their life. Thus, they were deprived of the taste of food and the food reward. The cats were then trained to find food behind the gate. They acquired this simple response very slowly and during the first weeks of training they often refused to eat the food reward. Moreover, they extinguished the response poorly. We conclude that the deprived cats did not originally perceive the food reward as attractive, but they were able to learn this, and they were poorly able to form both excitatory and inhibitory food conditioned connections. PMID- 10688062 TI - Bcl-w expression is increased in brain regions affected by focal cerebral ischemia in the rat. AB - Proteins of the bcl-2 family are important regulators of apoptosis in many tissues of the embryo and adult and may play a role in cell death following stroke. The recently isolated bcl-w gene encodes a pro-survival member of the bcl 2 family, which is widely expressed. However, it is not known whether bcl-w plays a role in determining cell survival after cerebral ischemia. Using Western blot analysis and immunocytochemistry, regional bcl-w protein expression was studied in rat brain 2, 6, 24 and 72 h following 20 min temporary middle cerebral artery occlusion (MCAO). Focal cerebral ischemia increased bcl-w protein expression within the caudate putamen and parietal cortex, as well as causing milder increases within frontal cortex. Immunocytochemically bcl-w was expressed within neurons (frontal and parietal cortex) and glia (caudate putamen) 24 h after MCAO. These data suggest that bcl-w could play a role in determining cell survival after cerebral ischemia. PMID- 10688063 TI - The practice of radiology is changing rapidly in many areas. PMID- 10688064 TI - CT diagnosis of acute flank pain from urolithiasis. AB - The use of noncontrast helical CT (NHCT) to assess patients with acute flank pain and hematuria for potential urinary tract stone disease was first reported in 1995. After several years of experience with the technique, sensitivity and specificity of NHCT has proven to be better than intravenous urography for evaluating ureteral stones. NHCT imaging findings for urinary calculi and the differential diagnosis are discussed in this article. Various extraurinary diseases found while using NHCT in searching for stone disease are addressed and illustrated. As experience with the use of NHCT has increased, clinicians have broadened the indications for this technique, which has a lower charge than standard CT, beyond the specific evaluation of urinary colic. This indication creep has increased the number of NHCT examinations ordered. It has also reduced the rate of stone positivity and increased the diagnostic yield for extraurinary disease. PMID- 10688065 TI - Targeted helical CT of the acute abdomen: appendicitis, diverticulitis, and small bowel obstruction. AB - CT, especially helical CT, provides a fast and reliable modality for evaluation of the patient presenting with acute abdominal pain. Helical CT can provide an accurate diagnosis in the majority of patients and has found great utility in the evaluation of acute gastrointestinal emergencies, including acute appendicitis, diverticulitis, and small bowel obstruction. This article reviews proper helical CT technique, diagnostic imaging findings, and pitfalls of interpretation in evaluation of these acute abdominal disorders. PMID- 10688066 TI - CT and MR diagnoses of intestinal ischemia. AB - CT and MR imaging have an important role in establishing the diagnosis of mesenteric ischemia. However, without specific signs such as thromboembolism in the mesenteric vessel, intramural or portal venous gas, and the absence of bowel wall enhancement, mesenteric ischemia can be confused with inflammatory or neoplastic gastrointestinal diseases. Arterial or venous occlusion or low-flow state are the main direct causes of mesenteric ischemia. Delayed diagnosis in equivocal cases can be avoided through an understanding of the patholophysiological aspects of mesenteric ischemia as they occur in a variety of other conditions, including: thromboembolism, bowel obstruction, neoplasm, vasculitis, inflammatory diseases, trauma, and drug or radiation therapy. PMID- 10688067 TI - Acute cholecystitis: CT findings. AB - Some patients with acute cholecystitis may have symptoms suggestive of an abscess or other intra-abdominal inflammation and, therefore, may be referred for a CT of the abdomen. This report reviews the pathophysiology, clinical presentation, and CT findings of acute cholecystitis (gallstones, wall thickening, distention, pericholecystic fluid, and pericholecystic stranding). Pitfalls and complications of the diagnosis are discussed. Those scenarios where CT may prove superior to ultrasound or hepatobiliary scintigraphy are highlighted. PMID- 10688068 TI - Assessment of acute abdominal pain in the pregnant patient. AB - The request to image a pregnant patient with abdominal pain often leads to concern and frustration for the referring clinician as well as the radiologist. In this report we will review the basic principles of radiation safety when imaging the pregnant woman, consider the diagnostic possibilities for the causes of abdominopelvic pain, and discuss the available imaging modalities to provide a basis for tailoring an imaging plan to the individual patient. PMID- 10688069 TI - Acute female pelvic pain: ultrasound evaluation. AB - Ultrasound has become a valuable primary imaging tool in the assessment of acute pelvic pain in women, both for diagnosis and for assessment of complications. Although ultrasound is an established imaging tool for gynecologic diseases, it is also a useful modality for assessing nongynecologic disorders that cause acute pelvic pain, such as diverticulitis and urinary tract calculi. These are important differential diagnoses in women with acute pelvic pain, and sonologists are not always expert in their diagnosis. This article reviews the gamut of conditions that can cause acute pelvic pain in women. The usual gynecologic causes are included, such as ectopic pregnancy, but also considered are conditions such as diverticulitis, appendicitis, and incarcerated hernia, which are important differential considerations. PMID- 10688070 TI - Adjuvant chemotherapy for T3 prostate cancer: not ready for prime time. PMID- 10688071 TI - Intravesical therapy for bladder cancer. PMID- 10688072 TI - Current evidence for the role of combined androgen suppression and radiation in the treatment of adenocarcinoma of the prostate. PMID- 10688073 TI - Comparison of expressed prostatic secretions with urine after prostatic massage- a means to diagnose chronic prostatitis/inflammatory chronic pelvic pain syndrome. AB - OBJECTIVES: To compare the analysis of urine after prostatic massage (VB3) with expressed prostatic secretions (EPS) to assess the significance of leukocyte analysis in VB3 and to give a first hint of the diagnosis of inflammatory chronic pelvic pain syndrome (CPPS) when EPS cannot be obtained. METHODS: Three hundred twenty-eight men (mean age 38 years, range 18 to 70) with expressible prostatic secretions were investigated. EPS were stained using the Papanicolaou stain and analyzed for leukocytes per high power field (HPF) (x1000). Additionally, identical aliquots of first voided urine (VB1), midstream urine (VB2), and VB3 were centrifuged, stained (Papanicolaou), and analyzed for leukocytes (x400). Patients with increased numbers of leukocytes in VB1 and VB2 (2 or more per x400) were excluded. For statistical analysis, Spearman's correlation coefficient for nonparametric tests was used. RESULTS: Of 180 men with less than 10 leukocytes per HPF in EPS, 178 (98.9%) had less than 10 leukocytes per view field in VB3. In 148 men with 10 or more leukocytes per HPF in EPS, 136 (91.9%) also had elevated leukocyte counts in VB3. The presence of elevated leukocytes in VB3 predicted the presence of increased leukocytes in EPS with a high certainty: 91.9% sensitivity, 98.9% specificity, and 95.7% accuracy, with a positive and negative predictive value of 98.6% and 93.7%, respectively. CONCLUSIONS: We conclude that the determination of leukocytes in VB3 is a feasible and reliable method compared with the analysis of EPS. However, although this association does not directly prove the significance of VB3 in those patients from whom no EPS can be obtained, we suggest this method be taken into account as an indirect indicator in the diagnosis of inflammation. PMID- 10688074 TI - Laparoscopic enterocystoplasty. AB - OBJECTIVES: To report the initial clinical experience with laparoscopic augmentation enterocystoplasty using the ileum, sigmoid, or right colon. METHODS: Three patients with functionally reduced bladder capacities due to neurogenic causes underwent laparoscopic enterocystoplasty: ileocystoplasty (n = 1), sigmoidocystoplasty (n = 1), and cystoplasty with cecum and proximal ascending colon (n = 1). In the last patient, a continent, catheterizable, ileal conduit with an umbilical stoma was also created. In all patients, bowel reanastomosis was performed by exteriorizing the bowel loop outside the abdomen through a 2-cm extension of the umbilical port site. Creation of a large cystotomy, mobilization of the appropriate bowel segment, and the circumferential enterovesical anastomosis were all performed intracorporeally by laparoscopic techniques. RESULTS: The operative times were 5.3, 8, and 7 hours. All three laparoscopic enterovesical anastomoses were watertight, without postoperative urinary extravasation. The hospital stay was 7, 5, and 4 days. CONCLUSIONS: Laparoscopic enterocystoplasty is feasible, safe, and efficacious and appears to be an attractive alternative to open enterocystoplasty. Various bowel segments can be used as with open surgery, including creation of a continent, catheterizable stoma. Although further technical refinements will undoubtedly occur, even at this early stage, it is clear that the technical steps of an enterocystoplasty can be satisfactorily and effectively performed laparoscopically. PMID- 10688075 TI - Double-blind randomized comparison of single-dose ciprofloxacin versus intravenous cefazolin in patients undergoing outpatient endourologic surgery. AB - OBJECTIVES: To compare the efficacy of single-dose oral ciprofloxacin with intravenous cefazolin as a prophylactic agent in patients undergoing outpatient endourologic surgery. METHODS: One hundred patients were enrolled in a double blind, randomized study to receive either ciprofloxacin (500 mg) or cefazolin (1 g) before surgery. A postoperative clinical evaluation and urine cultures were performed 5 to 10 days after surgery. Patients undergoing ureteral stent insertion or exchange, ureteroscopy, bladder biopsy, retrograde pyelography, collagen injection, and internal urethrotomy were included. RESULTS: Postoperative urinary tract infection occurred in 7 (9.1%) of 77 patients, including 3 (8.1%) of 37 and 4 (10.0%) of 40 of those who received ciprofloxacin and cefazolin, respectively (P = 0.77). There were no episodes of sepsis, and no patient with infection required hospitalization. The total cost associated with the administration of prophylactic antibiotics in the study population was $3657 less in those 50 patients who received ciprofloxacin than in the 50 patients who received cefazolin. CONCLUSIONS: A single oral dose of ciprofloxacin in patients undergoing outpatient endourologic surgery was equally effective as cefazolin in preventing postoperative urinary tract infection, but was associated with markedly lower overall costs. PMID- 10688076 TI - Detecting urethral and prostatic inflammation in patients with chronic prostatitis. AB - OBJECTIVES: Diagnosis of urethral and prostatic inflammation can represent a challenge. We compare the accuracy of diagnostic methods for detecting inflammation in lower urinary specimens/samples. METHODS: A standardized protocol was used to evaluate urethral smear, first-void urine (VB1), midstream urine (VB2), expressed prostatic secretions (EPS), and postmassage urine (VB3) in urologic patients with no symptoms or signs of urethritis who were attending our prostatitis clinic. RESULTS: Of 235 subjects, 60 (26%) had leukocytes detected by the Gram-stained urethral smear, 44 (18%) by the VB1, and only 14 (6%) by the VB2. Compared with the urethral swab, VB1 had 0% to 22% sensitivity and 81% to 98% specificity, and VB2 had 8% to 11% sensitivity. Of 83 subjects with prostatic inflammation, the EPS detected 63 (76%) and the VB3 detected 68 (82%). CONCLUSIONS: VB1 or VB2 examinations had low sensitivity for detecting urethral inflammation. Examining both the EPS and VB3 proved best for detecting prostatic fluid inflammation. Combining the urethral smear with lower urinary tract localization ("four-glass test") represents an optimal approach for detecting urethral and prostatic inflammation. PMID- 10688077 TI - Management of shotgun injuries to the pelvis and lower genitourinary system. AB - OBJECTIVES: Shotgun injuries are rare, with the extent of injury best determined at time of surgical exploration. There are no defined workup or management guidelines for patients with shotgun injuries to the genitourinary system. Injuries are usually treated on an individual basis. This study was conducted to determine the management and extent of genitourinary tract injuries in 10 patients with shotgun injuries to the pelvis during a 6-year interval. METHODS: Between September 1990 and December 1996, 140 patients were treated for firearm injuries to the lower genitourinary tract, of which 10 were secondary to shotgun blasts. We performed a retrospective hospital and clinic chart review and telephone interview to assess organs injured, initial treatment, follow-up surgeries, mortality, and erectile function. RESULTS: Mean patient age was 20 years at the time of the injury. The mean follow-up was 4 years (range 1 to 7). Two patients died, both with major vascular injuries, one in the operating room and the other 1 week later from sepsis. Eight patients underwent radiographic examinations (1 intravenous urogram and 7 urethrocystograms). The bladder was injured in 5 patients, 2 with concomitant complete posterior urethral transection. Of the 5 patients without bladder injury, one had an incomplete penile urethral injury and one had a complete bulbar urethral transection. The initial management consisted of repairing nongenitourinary injuries in 8 cases (80%), most commonly involving injuries to the rectum and small bowel. All patients were treated operatively, including 8 who required laparotomy and 4 who required suprapubic cystotomy. A total of four urethral injuries were noted. Subsequent reconstructive surgeries included two urethroplasties and one permanent supravesical diversion for 3 patients with extensive urethral loss. Erectile dysfunction was present in 3 of 6 patients available for telephone interview. CONCLUSIONS: Shotgun injuries involving the lower genitourinary tract are associated with significant soft tissue injury and morbidity. Death usually results from major associated vascular injuries. All hemodynamically stable patients should undergo retrograde urethrograms and cystograms to evaluate possible urethral and bladder injuries. Open primary repair should be attempted for distal urethral, testicular, and corporal injuries. Delayed repair with staged urethral reconstruction should be reserved for patients with extensive loss of urethral tissue. Impotence is common in patients with extensive perineal injuries. PMID- 10688078 TI - Ureteroscopic endopyelotomy in the treatment of patients with ureteropelvic junction obstruction. AB - OBJECTIVES: To investigate the effectiveness and morbidity of ureteroscopic endopyelotomy in adults with ureteropelvic junction (UPJ) obstruction. METHODS: Twenty-two patients (13 women, 9 men) with a mean age of 44 years (range 18 to 86) underwent retrograde ureteroscopic endopyelotomy in the treatment of primary (n = 18) or secondary (n = 4) UPJ obstruction. All procedures were performed using a 6F to 8.5F semirigid ureteroscope with either a 3F electrocautery probe (n = 16) or a 365-microm holmium laser fiber (n = 6). Postoperatively, a tapered 14/7F endoureterotomy stent (n = 11) or standard 7F to 8F double pigtail stent (n = 11) was left in place for 6 to 7 weeks. Radiographic follow-up was obtained using intravenous urography or renal scintigraphy. RESULTS: With a median follow up of 20.5 months, the success rate was 82% (18 of 22 patients). Follow-up of at least 6 and 12 months was available in 21 (95%) and 17 (77%) of 22 patients, respectively. The mean operative duration was 63 minutes, and all but 1 patient was hospitalized for less than 24 hours. No bleeding complications or other serious morbidity were encountered. No difference in treatment outcome was found on the basis of the size of the stent placed postoperatively, the incision type (cautery versus laser), or the etiology of the obstruction. CONCLUSIONS: Ureteroscopic endopyelotomy is an effective, minimally invasive treatment option for patients with primary or secondary UPJ obstruction. PMID- 10688079 TI - Mechanical percussion inversion can result in relocation of lower pole stone fragments after shock wave lithotripsy. AB - OBJECTIVES: To determine whether mechanical percussion combined with inversion (MPI) therapy and forced diuresis can move stone fragments out of the lower pole of the kidney. METHODS: Twelve patients with lower pole residual stone fragments at least 2 weeks after shock wave lithotripsy were treated using the following protocol. Eleven patients received 20 mg of furosemide before MPI therapy. Patients were treated in the prone Trendelenberg position on a pivoting stretcher and given 10 minutes of percussion over the flank using a mechanical chest physiotherapy percussor. Stone location was documented with plain abdominal radiographs before, immediately after, and 2 weeks following MPI therapy. Voided urine was strained immediately after MPI therapy and throughout the study period. RESULTS: Abdominal radiographs before and after treatment demonstrated movement of fragments out of the lower pole in 11 patients. In 8 patients, the lower pole appeared entirely clear of fragments on the immediate post-treatment film. Four patients passed stone fragments in their first voided urine. Ten patients passed stone fragments during the 2-week follow-up period. CONCLUSIONS: MPI therapy combined with diuresis can effectively mobilize stone fragments out of the lower pole calyces and appears to aid in the passage of fragments. PMID- 10688080 TI - Short-term complications of pubovaginal sling procedure for genuine stress incontinence in women. AB - OBJECTIVES: To evaluate the complication rate intraoperatively and within the first 30 postoperative days of the pubovaginal sling procedure. METHODS: From January 1992 to September 1996, we prospectively analyzed 90 women with type II and III genuine stress incontinence (age 38 to 84 years, average Valsalva leak point pressure 57.5 cm H2O) who underwent the pubovaginal sling procedure at our institute. Sixty percent of patients had no previous surgical treatment for their incontinence. Thirty-three percent of our patients have significant comorbidity (chronic obstructive lung disease, diabetes, coronary artery disease, peripheral vascular disease). RESULTS: The complication rate within the first 30 postoperative days was 19%, which included pneumonia (1.1%), deep venous thrombosis (1.1%), urinary retention (3.3%), wound infection (7.7%), intraoperative bladder laceration (3.3%), urinary tract infection (1.1%), and superficial thrombophlebitis (1.1%). A similar complication rate was noted among our patients with no previous surgical treatment for their incontinence. CONCLUSIONS: The complication rate of our prospective series of pubovaginal sling procedures was comparable to that of the other surgical procedures for genuine stress incontinence reported in the literature. We conclude that even though the pubovaginal sling procedure is a relatively complex surgery, in view of its satisfactory long-term success rate reported in the recent literature, pubovaginal sling procedure should also be considered a primary surgical treatment for genuine stress incontinence in a selected population of women. PMID- 10688081 TI - Distal ureteral regeneration after radical transurethral bladder tumor resection. AB - OBJECTIVES: Radical, full-thickness resection of the bladder wall and overlying bladder tumor is a management option in highly selected patients with muscle invasive bladder cancer. The consequences of iatrogenic ureteral injury in patients whose tumor involves the ureteral orifice and intramural ureter have not been reported. This report details an experience with 4 patients who underwent full-thickness resection of the hemi-trigone and distal ureter as treatment for muscle-invasive bladder tumors. METHODS: Between August 1995 and February 1999, 4 patients with T2 transitional cell cancer involving the bladder base and hemi trigone underwent radical transurethral resection of bladder tumor (TURBT), defined as resection of the tumor and bladder wall into the perivesical fat as primary tumor management. Six weeks later, the patients underwent a restaging TURBT to assess the pathologic response and status of the distal ureter. Patients were then followed up at regular intervals for the development of hydronephrosis and/or upper tract complications. RESULTS: Regeneration of the distal ureter was noted at 6 weeks in all patients. At a mean follow-up of 24 months, no patient had developed evidence of upper tract deterioration. All patients remained without evidence of tumor recurrence. CONCLUSIONS: This experience suggests that iatrogenic injury to the distal ureter during radical transurethral resection of tumor involving the hemi-trigone does not result in long-term distal ureteral damage. Involvement of the hemi-trigone by tumor does not appear to be a contraindication to radical TURBT. PMID- 10688082 TI - Modified Pereyra bladder neck suspension in patients with intrinsic sphincter deficiency and bladder neck hypermobility: patient satisfaction with a mean follow-up of 4 years. AB - OBJECTIVES: To determine the long-term success rate for the modified Pereyra bladder neck suspension and to identify preoperative characteristics that create differences in surgical outcome. We attempted retrospectively to separate those patients with what we now recognize was significant intrinsic sphincter deficiency (ISD) before routine use of Valsalva leak point pressures (VLPPs) was available. METHODS: The charts and videourodynamic reports of 208 patients who underwent a modified Pereyra bladder neck suspension from June 1988 to June 1996 were reviewed, and survey questionnaires were mailed to all patients. All videourodynamic study reports and charts were reviewed to identify those with what we now recognize was significant ISD and compare them with a group that we believed had more pure descent problems. RESULTS: A total of 135 patients or 65% of the population responded. The mean time after surgery was 4.14 years. At the follow-up survey, 14% reported no leakage at all, 42% reported very little or mild leakage, 38% reported moderate leakage, and 6% reported severe leakage. Fifty-three percent of patients continued to wear pads. Seventy-nine percent reported improvement in their leakage compared with the preoperative state, and 69% were satisfied with the results. When patients with preoperative ISD were compared with patients with pure bladder neck hypermobility, the ISD group had more leakage and less improvement after surgery than patients with bladder neck hypermobility. CONCLUSIONS: With an average follow-up of greater than 4 years, most women continued to leak with symptoms of stress urinary incontinence. Even though 79% reported improvement over their preoperative condition and 69% were satisfied, the results were disappointing. Patients with significant ISD had a worse outcome (2.6% dry) than patients with pure bladder neck hypermobility (20% dry). Given the above data, significant ISD is a contraindication for a modified Pereyra transvaginal needle suspension, and these data cast further doubt on the ability of the modified Pereyra needle suspension to consistently cure even anatomic incontinence. PMID- 10688083 TI - Comparison of the monoclonal UBC-ELISA test and the NMP22 ELISA test for the detection of urothelial cell carcinoma of the bladder. AB - OBJECTIVES: To compare the diagnostic value of two enzyme-linked immunosorbent assay (ELISA) tests, the nuclear matrix protein 22 (NMP22) test and a newly developed urinary bladder cancer (UBC) test, in patients having symptoms suggestive of urothelial cell carcinoma (UCC) and patients under follow-up after transurethral resection (TUR). METHODS: Two hundred forty patients with a mean age of 65.8 years (range 22 to 92) were included in this retrospective study. The tests were performed on previously frozen urine samples. Eighty-one patients had symptoms suggestive of bladder cancer and 159 patients were being followed up after complete TUR of UCC. Voided urine was evaluated by the NMP22 test and the monoclonal UBC-ELISA test, which traces cytokeratins 8 and 18. All patients underwent subsequent cystoscopy and biopsy evaluation of any suspicious lesion. The cutoff levels for bladder cancer positivity were 10 U/mL for the NMP22 test and 12 microg/L for the UBC test. RESULTS: In the 54 patients with histologically proved UCC, the NMP22 test had a sensitivity of 55.5% and the UBC test a sensitivity of 64.8%. According to the histologic stages, the sensitivity of NMP22 was 51.7% in pTa tumors, 46.1% in pT1, and 70% in pT2 or higher tumors; the sensitivity of UBC was 62.1% in pTa, 53.8% in pT1, and 80% in pT2 or higher tumors. For histologic grades 1 to 3, the sensitivity was 50%, 50%, and 68.7% for NMP22 and 66.6%, 60%, and 68.7% for UBC, respectively. The specificity was 79% and 92% for NMP22 and UBC, respectively. CONCLUSIONS: The monoclonal UBC-ELISA test is superior to the NMP22 test in both sensitivity and specificity. Nevertheless, neither test can replace cystoscopy. PMID- 10688084 TI - Urinary NMP22 and renal cell carcinoma. AB - OBJECTIVES: To demonstrate the incidence of positive urinary nuclear matrix protein 22 (NMP22) values associated with renal cell carcinoma (RCC) of the kidney. Currently, urinary NMP22 is used to detect recurrent transitional cell carcinoma of the bladder. METHODS: From May 1997 to March 1998, urinary NMP22 values were obtained from 65 patients who had undergone either computed tomography scanning of the abdomen or renal ultrasound. Of the 65, 32 presented with solid renal masses. These patients underwent radical or partial nephrectomy; subsequent pathologic examination revealed that 30 had RCC. Two patients had oncocytomas. The remaining 33 patients presented with blunt abdominal trauma or abdominal pain or for follow-up for a urologic problem (a kidney stone or benign renal cyst). These patients had no evidence of renal malignancy on imaging and were used as controls. RESULTS: The urinary NMP22 values of patients with RCC were significantly higher than the values found in the control group (13.69 +/- 8.40 U/mL versus 3.03 +/- 2.70 U/mL, P <0.0078). Of the 30 patients with RCC, 12 (40%) had positive urinary NMP22 values of 10 U/mL or above. Chi-square analysis revealed a significant difference between the NMP22 values of the two groups (P <0.005). There were two false-positive NMP22 values in the 35 control patients. CONCLUSIONS: Urinary NMP22 values used to detect transitional cell carcinoma in individuals with previously diagnosed bladder cancer should be more broadly evaluated, since elevations have also been found to occur in RCC of the kidney. This finding may result in an increase in the incidental discovery of RCC. Future, more specific, NMP assays may hold promise for the detection of RCC. PMID- 10688085 TI - Clinical outcome 1 year after transurethral vaporization and resection of the prostate. AB - OBJECTIVES: To evaluate the 1-year safety and effectiveness of transurethral vaporization of the prostate (TUVP) compared with transurethral resection of the prostate (TURP) in alleviating outflow obstruction. METHODS: Two experienced surgeons performed 26 TUVPs and 28 TURPs. The intraoperative blood loss was measured by photometry and fluid absorption by the ethanol method. The treatments were evaluated by means of the International Prostate Symptom Score (IPSS), quality-of-life score, transrectal ultrasound, prostate-specific antigen level, urinary flow rate, and postvoid residual urine volume. RESULTS: After TUVP, the median IPSS decreased from 22 to 4.5 and the quality-of-life score from 4.5 to 1.5. The corresponding data for TURP were from 25 to 5.5 and from 4.0 to 1.0. The median urinary flow rate increased from 4 to 10 mL/s after TUVP and from 2 to 11 mL/s after TURP. The postvoid residual urine volume decreased to 35% (TUVP) and 15% (TURP) of the preoperative volume. The blood loss was larger during TURP (P <0.04), but complications during follow-up were more frequent after TUVP (P <0.02). Patients with fluid absorption during surgery had a lower quality-of-life score at follow-up (P <0.02) and tended to have a smaller reduction in prostate size (to 79%) than those without absorption (to 67% of baseline). CONCLUSIONS: TUVP and TURP were both effective in alleviating outflow obstruction, but the outcome appeared to be slightly better for TURP. PMID- 10688086 TI - Incidence of acute myocardial infarction and cause-specific mortality after transurethral treatments of prostatic hypertrophy. AB - OBJECTIVES: Transurethral resection of the prostate (TURP) is associated with a higher long-term mortality than open prostatectomy which has been ascribed to a higher incidence of acute myocardial infarction (AMI). To assess the possible excess risk associated with TURP, we studied the incidence of AMI and the cause specific mortality in patients treated with TURP and transurethral microwave thermotherapy (TUMT). METHODS: Patients treated for benign prostatic hypertrophy at a university hospital (888 patients with TURP and 478 with TUMT) were monitored during an average follow-up period of 3.9 years. The incidence of AMI and the causes of death were compared with those in the general population. RESULTS: Both treatments were followed by a higher incidence of AMI than in the general population, in particular from 2 years or more after treatment (standardized morbidity ratio 1.50, 95% confidence interval [CI] 1.14 to 1.93). The long-term mortality from all causes was increased in patients younger than 75 years of age when undergoing any of the treatments (standardized mortality ratio [SMR] 1.16, 95% CI 0.97 to 1.39), in particular, death from cardiovascular diseases (SMR 1.25, 95% CI 0.95 to 1.60) and tumors (SMR 1.54, 95% CI 1.14 to 2.03). CONCLUSIONS: The similarity of the results for TURP and TUMT suggests that the prostatic enlargement rather than the treatment is associated with cardiovascular disease. PMID- 10688087 TI - Role of Viagra after radical prostatectomy. AB - OBJECTIVES: To determine whether the response to sildenafil citrate (Viagra) in patients with erectile dysfunction after radical prostatectomy was influenced by the presence or absence of neurovascular bundles, the interval from surgery to the initiation of drug therapy, and the dose of the drug. METHODS: Baseline and follow-up data from 91 patients presenting with erectile dysfunction after radical prostatectomy were obtained. The patients were stratified according to the type of nerve-sparing (NS) procedure: bilateral NS, unilateral NS, and non NS. They were interviewed using the Cleveland Clinic Post Prostatectomy (CCPP) questionnaire and the International Index of Erectile Function (IIEF) questionnaire. RESULTS: The presence or absence of the neurovascular bundles influenced the ability to achieve vaginal intercourse. In the patients who had undergone bilateral NS, 71.7% (38 of 53) responded; in those with unilateral NS, 50% (6 of 12) responded; and in those with non-NS, 15.4% (4 of 26) responded. The IIEF questionnaire confirmed the quality of the positive responses, with significant improvements in response to question 3 (frequency of penetration), question 4 (frequency of maintenance of erection), and question 7 (satisfaction with intercourse). The magnitude of improvement in responses was higher in the bilateral NS group than in the unilateral NS and non-NS groups (P <0.05). When the data of the 48 positive responders were analyzed, no difference in the response rate was found when the interval from surgery to drug therapy was stratified by the following three intervals: 0 to 6 months (44%), 6 to 12 months (55%), and greater than 12 months (53%). Of the positive responders, 14 (29.1%) required the 50-mg dose, and 34 (70.9%) required the 100-mg dose. The most common side effects were transient headaches (28.6%), flushing (21.9%), dizziness (8.8%), dyspepsia (6.5%), and nasal congestion (5.4%), with an increase in the incidence of headaches seen at the higher dose (P = 0.04). CONCLUSIONS: Successful treatment of erectile dysfunction with sildenafil citrate after radical prostatectomy depends on the presence of the neurovascular bundles. Our data suggest that the response to sildenafil is not related to the interval between the surgery and initiation of drug therapy but is related to the dose. PMID- 10688088 TI - Prostate cancer biochemical recurrence stage for stage is more frequent among African-American than white men with locally advanced but not organ-confined disease. AB - OBJECTIVES: To determine whether outcome differences between African-American men (AAM) and white men with prostate cancer (PCa) will still be present if we control for stage in a large cohort of men. It is well established that AAM have a worse outcome from PCa than white men. METHODS: We examined 848 consecutive patients who underwent radical prostatectomy at Wayne State University, Karmanos Cancer Institute, between 1991 and 1995. The mean follow-up was 34 months (range 1.5 to 75). We included men with Gleason score 7 (4 + 3) with those men with Gleason score 8 to 10 for racial/ethnic comparisons. RESULTS: AAM and white men diagnosed with organ-confined PCa demonstrated similar prostate-specific antigen (PSA) levels, Gleason grade, and biochemical recurrence. However, AAM diagnosed with non-organ-confined disease demonstrated higher PSA levels and a higher incidence of recurrence than did white men with non-organ-confined disease. There was a trend toward AAM having a greater proportion of high-grade lesions than white men when PCa was not organ confined. The evidence suggests that the difference in recurrence among AAM versus white men is based on pretreatment PSA, grade, extracapsular extension, and positive surgical margins. Seminal vesicle invasion predicted a worse prognosis equally for both AAM and white men. CONCLUSIONS: A difference in biochemical recurrence was not detected between AAM and white men with organ-confined PCa after radical prostatectomy. PSA was higher in AAM than in white men with pathologically locally advanced PCa, and the biochemical recurrence was greater. AAM had a greater percentage of high Gleason grade lesions compared with white men, and this difference approached statistical significance. We hypothesize that AAM have a more rapid growth rate of PCa, which may be responsible for these clinical findings. Further investigations of the biology of PCa are needed to understand these findings. PMID- 10688089 TI - Novel image analysis of corpus cavernous tissue in impotent men. AB - OBJECTIVES: To objectively evaluate the contents of corpus cavernous tissue in impotent men using an automated novel image analysis system. METHODS: Thirty three impotent men and 2 normal potent men (controls) underwent corpus cavernous biopsies. The procedures were performed using a Biopty gun under local anesthesia. The obtained specimens were stained with Masson's trichome technique, and the collagen fiber contents were evaluated by a computerized morphometric analysis method. In addition, we estimated the intraobserver and interobserver reliability of this automated image analysis system. RESULTS: No major complication was noted during or after the biopsies. Of the 33 impotent patients, 3 were diagnosed as having psychogenic, 11 as having arteriogenic, 13 as having venogenic, 1 as having neurogenic, and 5 as having idiopathic impotence. The collagen fiber percentages in the 35 men were as follows: normal potent: 48.2% +/ 1.4%, psychogenic 55.2% +/- 11.6%, arteriogenic 73.2% +/- 4.4%, venogenic 66.5% +/- 4.2%, neurogenic 76.9%, and idiopathic 77.4% +/- 4.2%. Significant differences were found between the normal potent and arteriogenic groups (P <0.05) and between the normal potent and idiopathic groups (P <0.05). Patients older than 60 years had a higher collagen fiber content (70.4% +/- 3.7%) than those younger than 50 years old (58.6% +/- 5.2%). The interobserver and intraobserver variances were both negligible for this automated image analysis system. This method significantly reduced the amount of variation introduced by the intra-rater reliability of a technician compared with the manual method. CONCLUSIONS: The present automated image analysis system is believed to be a reliable, accurate quantitative measurement tool for studies of penile tissue. Cavernous biopsy is a rapid, safe, and representative modality to study penile disease. An increase in cavernous collagen fibers (or corporal fibrosis) is considered an important factor in impaired erectile function. PMID- 10688090 TI - Clinical study of SS-cream in patients with lifelong premature ejaculation. AB - OBJECTIVES: To investigate the clinical efficacy of SS-cream, the topical agent made from the extracts of nine natural products for the treatment of premature ejaculation, we performed a double-blind, randomized, placebo-controlled Phase III clinical study of patients with lifelong premature ejaculation in three medical centers. METHODS: One hundred six patients (mean age 38.7 +/- 0.61 years) completed this study. The ejaculatory latency measured by stopwatch and sexual satisfaction ratio of both partner and patient were investigated twice in the screening period and once after each treatment (1 placebo 0.20 g and 5 SS-cream 0.20 g for a total of six treatments). Patients were instructed to apply the cream on the glans penis 1 hour before sexual intercourse in a double-blind randomized fashion. Clinical efficacy was compared with the prolongation of ejaculatory latency and improvement of the sexual satisfaction ratio before and after each treatment. RESULTS: In the screening period, the mean ejaculatory latency was assessed at 1.37 +/- 0.12 minutes, and neither the patients nor their partners were satisfied with their sexual lives. After treatment, the mean ejaculatory latency was prolonged to 2.45 +/- 0.29 minutes in the placebo group and 10.92 +/- 0.95 minutes in the SS-cream group. The clinical efficacy of placebo and SS-cream as judged by an ejaculatory latency time prolonged more than 2 minutes was 15.09% and 79.81%, respectively. The improvement of sexual satisfaction to a grade higher than effective was 19.81% and 82.19%, respectively, for placebo and SS-cream. Of 530 trials of SS-cream, 98 (18.49%) resulted in a sense of mild local burning and mild pain. No adverse effect on sexual function or partner and no systemic side effects were observed. CONCLUSIONS: According to these results, SS-cream is effective and safe in the treatment of premature ejaculation, with mild local side effects. PMID- 10688091 TI - Magnetic resonance imaging and magnetic resonance angiography before postchemotherapy retroperitoneal lymph node dissection. AB - OBJECTIVES: Retroperitoneal lymph node dissection (RPLND) after primary chemotherapy is an accepted therapeutic approach for metastatic nonseminomatous germ cell testicular cancer. Because of the intense desmoplastic reaction and adherence to venous and arterial walls, accurate imaging of the retroperitoneal vasculature and its relation to residual tumor is essential. We report our experience with magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA), including the recently developed technique of bolus-contrast MRA, in patients undergoing postchemotherapy RPLND. METHODS: Eighteen patients underwent MRI of the retroperitoneal region before RPLND. In addition to routine sequences, MRA was performed in 10 patients, including 8 with a three-dimensional technique using bolus intravenous MR contrast. Results were compared with intraoperative and pathologic findings. RESULTS: MRI and MRA provided detailed information on retroperitoneal vasculature and its relation to tumor, including multiple renal vessels (n = 5), duplex inferior vena cava (n = 1), left retroaortic renal vein (n = 2), and common iliac vein thrombus (n = 1). In all cases, bolus-contrast MRA provided unique information on the location and number of renal and lumbar arteries, in addition to information on the aorta and the mesenteric and iliac vessels. The origin and number of renal arteries were accurately identified in all patients by bolus-contrast MRA; 2 patients had supernumerary renal arteries discovered at RPLND that had not been identified on non-bolus-contrast MRI. CONCLUSIONS: Bolus-contrast three-dimensional MRA provides unique information on renal and lumbar vessels. The potential benefit of avoiding vascular injury during dissection should be prospectively evaluated. PMID- 10688092 TI - Extensive surgery on the trigone for complete ureteral duplication does not cause incontinence or voiding problems. AB - OBJECTIVES: To evaluate whether extensive trigonal surgery for duplicated kidneys is harmful for later bladder and urethral function. METHODS: Of 201 surgically treated children with kidney and ureteral duplication, 145 were followed up for at least 1 year. The mean follow-up was 5 years (range 1 to 15), and all patients were at least 7 years old at the date of their last follow-up visit. Trigone surgery was performed in 105 children; bilateral trigonal surgery in 26, unroofing in 25, and total excision in 5. On all later consultations, the presence of infection, voiding habits, continence pattern, and ultrasound findings for residual urine volume and kidney function were noted. Children with recurrent urinary tract infection or dysfunctional voiding for more than 2 years underwent a urodynamic examination. RESULTS: Nine children, of whom five were boys, had nocturnal enuresis only. Eight patients had day and nighttime wetting. Seven of the 8 patients had recurrent urinary infections; urodynamic evaluation revealed a high compliance (with residual urine) in three of these children and four had detrusor instability. One girl had an irregular bladder neck, with stress incontinence. All reflux, whether surgically or conservatively treated and also three of four occurring de novo, disappeared within 1 year after surgery. In the group without voiding dysfunctions, seven cystitis and five pyelonephritis attacks occurred. CONCLUSIONS: Neither extensive trigonal surgery nor pre existing trigonal deformation by ureteroceles provokes later bladder dysfunction. PMID- 10688093 TI - Office pediatric urologic procedures from a parental perspective. AB - OBJECTIVES: To determine the parent perception of discomfort (PPD) in children receiving local anesthesia for the lysis of labial adhesions, meatotomy, and newborn circumcision; the parents' perceived outcome of these procedures; and the overall satisfaction of parents when these procedures are performed in the office. METHODS: A confidential phone survey was given to 99 parent participants whose children had received local anesthesia for the lysis of labial adhesions (n = 14), meatotomy (n = 28), or newborn circumcision (n = 57). Parents in the labial adhesion and meatal stenosis groups were asked to rate their child's level of discomfort (PPD) during the procedure as mild (1), moderate (2), or severe (3), and those in the circumcision group were asked to use the same scale to rate their child's discomfort after the procedure. Parents were also asked whether they thought the procedure was successful and whether they were satisfied with their decision to have it done in the office using local anesthesia. RESULTS: Among the labial adhesion, meatotomy, and circumcision groups, the mean PPD +/- SD was 1.64 +/- 0.75, 1.54 +/- 0.69, and 1.21 +/- 0.53, respectively. The observed procedures, lysis of labial adhesions and meatotomy, had a significantly higher PPD score (P = 0.005) than the unobserved procedure (circumcision). Parents reported good outcomes in 94 (94.9%) of 99 children, with 4 girls experiencing recurrent labial adhesions and 1 boy experiencing recurrent meatal stenosis. Overall, 95 (96.0%) of 99 parents stated that they were satisfied with their decision to have the procedure done in the office with local anesthesia. Of the remaining 4 parents, 3 parents in the circumcision group stated they would have preferred general anesthesia, and 1 parent in the labial adhesion group was undecided. CONCLUSIONS: The PPD is greater if the parent observes their child's procedure. However, office procedures using local anesthesia for the lysis of labial adhesions, meatotomy, and newborn circumcision are well accepted among the parent population. PMID- 10688094 TI - Acucise endopyelotomy. AB - INTRODUCTION: The evolution of minimally invasive therapy for ureteropelvic junction (UPJ) obstruction has culminated with the Acucise endopyelotomy. Antegrade endopyelotomy, laparoscopic pyeloplasty, and ureteroscopic endopyelotomy all offer excellent minimally invasive alternatives to open pyeloplasty, yet still represent more invasive techniques than the Acucise endopyelotomy in treating the obstructed UPJ. TECHNICAL CONSIDERATIONS: The Acucise endopyelotomy is a straightforward, efficacious, and safe procedure in the appropriate patient for treating UPJ obstruction. Under fluoroscopic guidance, the latest version of the Acucise allows the urologist to perform a retrograde pyelogram, position the Acucise catheter, make the incision, and place a ureteral stent, all over a single guide wire. In my experience, this latest technical modification has further simplified the procedure for the practicing urologist. CONCLUSIONS: In 2000, the Acucise endopyelotomy continues to represent an excellent minimally invasive option for all urologists who choose to perform endopyelotomies. PMID- 10688095 TI - Images in clinical urology. Cystic lymphangioma of the retroperitoneum. PMID- 10688096 TI - Images in clinical urology. Magnetic resonance imaging of primitive neuroectodermal tumor of the kidney. PMID- 10688097 TI - Absence of microsatellite instability in transitional cell carcinoma of the bladder. AB - OBJECTIVES: To investigate the prevalence of the microsatellite instability related to mismatch repair (MMR) gene defects using a panel of six microsatellite markers, as recommended by a recent workshop on microsatellite instability in colon cancer, because it is still unclear whether abnormalities in DNA MMR genes are involved in transitional cell carcinoma (TCC) of the bladder. METHODS: Three mononucleotide repeats (BAT26, TGFbetaRII, and BAX) were studied in 33 TCC samples and in four bladder cancer cell lines. Three dinucleotide repeats (D2S123, D5S346, and D17S250) were studied in 21 of these 33 TCC samples. RESULTS: No alteration was detected either in the 33 TCC samples analyzed or in the four bladder cancer cell lines (T24, J82, 647V, and 1207) studied. A difference between normal and tumor DNA was observed in only 1 of 21 tumor samples for D17S250. CONCLUSIONS: These data indicate that microsatellite instability is very uncommon in TCC of the bladder. PMID- 10688098 TI - Functional evaluation of Tadenan on micturition and experimental prostate growth induced with exogenous dihydrotestosterone. AB - OBJECTIVES: To evaluate the effect of Tadenan (TAD; Pygeum africanum extract) pretreatment on the micturition characteristics of conscious and anesthetized rats consequent to dihydrotestosterone (DHT) administration and to examine the influence of such treatment on the growth of the prostate. METHODS: Studies using 40 adult Sprague-Dawley male rats were performed during a 7-week period. These animals were treated with DHT 1.25 mg/kg subcutaneously dissolved in peanut oil and/or TAD 100 mg/kg orally dissolved in sesame oil, except for the controls, which received vehicle only. Rats were divided into four groups: group 1 (control), vehicle only; group 2, DHT administered during weeks 3 and 4; group 3, TAD pretreatment, administered during weeks 1 and 2, followed by the combined administration of DHT and TAD during weeks 3 and 4 and TAD only during weeks 5 to 7; and group 4, continuous TAD treatment for 7 weeks. Micturition of conscious rats was evaluated in metabolic chambers, and in anesthetized rats, cystometrograms were done at the end of 7 weeks. RESULTS: DHT or DHT plus TAD did not produce significant changes in the volume but did reduce the frequency of micturition. TAD given alone significantly increased the volume of micturition and the rate of urine production. Cystometrographic studies in anesthetized rats revealed that DHT produced micturition characteristics similar to obstruction. The DHT plus TAD and TAD pretreatment data showed no significant difference from controls, suggesting that in the presence of TAD, the effects of DHT were negated. The total prostate weight of DHT and DHT plus TAD pretreated rats increased, and in the TAD group, these values decreased to lower than controls; growth of the ventral lobes was suppressed in the presence of TAD. CONCLUSIONS: These results demonstrate that TAD pretreatment significantly reduces the "obstructive" effects of DHT on micturition, counteracts the hormone-induced enlargement of the prostate, and reduces prostate weight in the ventral but not the dorsal lobe. PMID- 10688099 TI - Penile implant success in patients with corporal fibrosis using multiple incisions and minimal scar tissue excision. PMID- 10688101 TI - Y2OK! Rollover uneventful for nuclear medicine. PMID- 10688100 TI - Ultrasound-guided brachytherapy: is it really better? PMID- 10688102 TI - 99mTc-human serum albumin scans in children with protein-losing enteropathy. AB - Protein-losing enteropathy (PLE) can be diagnosed scintigraphically using 99mTc human serum albumin (HSA) scans. METHODS: To evaluate the usefulness of this method in detecting enteric protein loss, we retrospectively reviewed the 99mTc HSA scans of 18 children presenting consecutively with PLE. RESULTS: Enteric 99mTc-HSA uptake was noted in 12 patients (8 boys, 4 girls) with a mean age of 7.4 y. Early dynamic images showed abdominal uptake that was most likely in the small bowel in 91% of the scans. Delayed images showed abnormal accumulation that was localized in the colon in 73% and in the small bowel in 27% of the scans. A 4 mo follow-up scan obtained in 3 patients showed reduced HSA uptake after a high protein, low-fat, medium-chain triglyceride oil-based diet and fat-soluble vitamins. Mean serum albumin, total protein, gammaglobulin, and calcium levels were significantly decreased. Ten patients (from 4 families) were diagnosed to have primary intestinal lymphangectasia. One patient had active Salmonella enterocolitis, and 1 had giardiosis. 99mTc-HSA was normal in the remaining 6 patients (3 boys, 3 girls) with a mean age of 3.5 y (range, 2-5 y). Mean serum albumin, total protein, gammaglobulin, and calcium levels were less decreased than those of the first group. Five of these patients had primary intestinal lymphangactesia (associated with infantile systemic hyalinosis in 1 patient). The remaining patient had normal duodenal biopsy, and the cause of protein loss remained unknown. CONCLUSION: The 99mTc-HSA scan is useful in the evaluation of children with PLE, especially those with severe hypoproteinemia and hypoalbuminemia, presumably reflecting a high rate of protein loss. PMID- 10688103 TI - Optimization of automated quantification of 123I-IBZM uptake in the striatum applied to parkinsonism. AB - Evaluation of therapies for parkinsonism by dopamine receptor SPECT requires a reproducible, optimized quantitation technique. This study presents a new, objective, automated technique for semiquantitative analysis of dopamine receptor density, as applied to the differential diagnosis of parkinsonism. METHODS: Dopamine receptor density measured by 123I-iodobenzamide (IBZM) SPECT was retrospectively analyzed in nonidiopathic parkinsonism (NIPS), in Parkinson's disease (PD), and in healthy volunteers (n = 19, 38, and 13, respectively). A mean template was created from coregistered control studies. Registration errors were assessed using studies with simulated binding deficits. Patient studies were registered to the mean template, and striatal binding was calculated from a corresponding map of 3-dimensional regions of interest (ROIs). The striatal binding ratio and deficits determined by voxelwise comparison with the normal template were investigated and tested with various 3-dimensional ROI sizes and positions. Separation of patient groups was determined by tscore after automatically processing all studies. Results were compared with manual ROI analyses. RESULTS: The automatic method was completely reproducible in 64 of 70 cases. The best diagnostic discriminator was the minimum binding ratio of the 2 striatal nuclei, with the following values: NIPS, 1.33+/-0.13; PD, 1.50+/-0.12; healthy volunteers, 1.49+/-0.08 (+/-SD). The deficit size from voxelwise analysis was: NIPS, 20.5+/-8.2 mL; PD, 9.5+/-8.3; healthy volunteers, 8.9+/-6.0 (+/-SD). The accuracy, measured by receiver operating characteristic areas, was 0.85+/ 0.05, 0.77+/-0.06, and 0.80+/-0.06 (+/-SE) for the optimal predictor (automated) and 2 blinded observers (manual), respectively. CONCLUSION: A new 3-dimensional, automated technique has been developed to semiquantitate receptor density that dramatically improves reproducibility. The optimal diagnostic discriminator of parkinsonism determined by the automatic technique has good accuracy compared with the manual technique. PMID- 10688104 TI - Frontotemporal decreases in rCBF correlate with degree of dysnomia in primary progressive aphasia. AB - Primary progressive aphasia (PPA) is an uncommon degenerative dementia characterized by gradual impairment of language function with initial sparing of the memory domain. Using semiquantitative 99mTc-hexamethyl propyleneamine oxime (HMPAO) brain SPECT as a measure of regional cerebral blood flow (rCBF), we investigated the relationship between reduced 99mTc-HMPAO uptake and the severity of dysnomia in PPA. METHODS: Seven right-handed patients with PPA had their dysnomia assessed by the Boston Naming Test (BNT), a subtest of the Boston Diagnostic Aphasia Examination. Neuroimaging studies, including 99mTc-HMPAO brain SPECT, CT, and MRI, were performed. Correlational analysis between reduced rCBF and BNT was performed. RESULTS: Brain SPECT showed a reduction in 99mTc-HMPAO uptake involving the frontal and temporal lobes in all 7 patients. CT and MRI showed mild to moderate cerebral atrophy in 4 patients. Low scores on the BNT correlated with low frontotemporal 99mTc-HMPAO (Spearman r = 0.97, P = 0.004) in the 5 patients with left-hemisphere involvement. CONCLUSION: Decreased rCBF to the frontotemporal region characterized the cerebral abnormalities associated with PPA. The finding of focal rCBF abnormalities in the right hemisphere of 2 right-handed women corroborates that PPA symptoms may arise from a "non-left dominant"-hemisphere degenerative process. Our results support the usefulness of rCBF SPECT imaging as a diagnostic aid in PPA. PMID- 10688105 TI - PET quantification of 5-HT2A receptors in the human brain: a constant infusion paradigm with [18F]altanserin. AB - [18F]altanserin has been used to label serotonin 5-HT2A receptors, which are believed to be important in the pathophysiology of schizophrenia and depression. The purpose of this study was to test the feasibility of a constant infusion paradigm for equilibrium modeling of [18F]altanserin with PET. Kinetic modeling with [18F]altanserin may be hampered by the presence of lipophilic radiometabolites observed in plasma after intravenous administration. METHODS: Eight healthy volunteers were injected with [18F]altanserin as a bolus (208+/-9 MBq [5.62+/-0.25 mCi]) plus constant infusion (65+/-3 MBq/h [1.76+/-0.08 mCi/h]) ranging from 555 to 626 min (615+/-24 min) after injection. PET acquisitions (10 20 min) and venous blood sampling were performed every 30-60 min throughout the infusion period. RESULTS: Linear regression analysis revealed that time-activity curves for both brain activity and plasma [18F]altanserin and metabolite concentrations stabilized after about 6 h. This permitted equilibrium modeling and estimation of V3' (ratio of specific uptake [cortical-cerebellar] to total plasma parent concentration after 6 h). Values of V3' ranged from 1.57+/-0.38 for anterior cingulate cortex to 1.02+/-0.39 for frontal cortex. The binding potential V3 (ratio of specific uptake to free plasma parent concentration after 6 h, using group mean f1) was also calculated and ranged from 169+/-41 for anterior cingulate cortex to 110+/-42 for frontal cortex. From 6 h onward, the rate of change for V3' and V3 was only 1.11+/-1.69 %/h. CONCLUSION: These results demonstrate the feasibility of equilibrium imaging with [18F]altanserin over more than 5 radioactive half-lives and suggest a method to overcome difficulties associated with lipophilic radiolabeled metabolites. The stability in V3 and V3' once equilibrium is achieved suggests that a single PET acquisition obtained at 6 h may provide a reasonable measure of 5-HT2A receptor density. PMID- 10688106 TI - 123I-interleukin-2 scintigraphy for in vivo assessment of intestinal mononuclear cell infiltration in Crohn's disease. AB - Activated mononuclear cells expressing interleukin-2 (IL2) receptors (IL2-Rs) heavily infiltrate the Crohn's disease (CD) gut wall. A new technique for the in vivo detection of tissue infiltrating IL2-R positive (IL2R+ve) cells was developed based on 123I-IL2 scintigraphy. The aim of this study was to investigate whether 123I-IL2 accumulates in the CD gut wall in different phases of the disease and to evaluate the specificity of 123I-IL2 binding to activated IL2R+ve cells infiltrating the gut wall. METHODS: Fifteen patients with ileal CD (10 active and 5 inactive) and 10 healthy volunteers were studied by 123I-IL2 scintigraphy. Six patients with active CD were studied before and after 12 wk of steroid treatment. After scintigraphy, patients were followed up for 29-54 mo. Ex vivo autoradiography was performed to determine specificity of 125I-IL2 binding to IL2R+ve cells. For bowel scintigraphy, 123I-IL2 (75 MBq) was injected intravenously and gamma camera images were acquired after 1 h. Bowel radioactivity was quantified in 64 regions of interest (ROIs). RESULTS: Autoradiography showed specific binding of 125I-IL2 to IL2R+ve mononuclear cells infiltrating the CD gut wall. Intestinal 123I-IL2 uptake assessed by the number of positive ROIs was higher in patients with active or inactive CD than in healthy volunteers (P < 0.0001 and P = 0.03, respectively) and positively correlated with the CD activity index (P = 0.01). 123I-IL2 intestinal uptake significantly decreased in patients with CD in steroid-induced remission (P = 0.03). A significant correlation was observed between the number of positive ROIs and time to disease relapse. CONCLUSION: 123I-IL2 accumulates in the diseased CD gut wall by specific binding to IL2R+ve cells, infiltrating the involved tissues. 123I-IL2 scintigraphy may be an objective tool for the in vivo assessment of intestinal activated mononuclear cell infiltration. PMID- 10688107 TI - Radiation synovectomy using 165Dy ferric-hydroxide and oxidative DNA damage in patients with different types of arthritis. AB - Radiation synovectomy is an effective treatment for chronic synovitis refractory to pharmacological treatment in patients with rheumatoid or seronegative arthritis. Concerns persist about possible radiation-induced cytogenetic damage after radiation synovectomy leading to recommendations to use this technique only in the elderly. Micronucleus (MN) frequency in lymphocytes and urinary excretion of 8-hydroxy-2'-deoxyguanosine (8OHdG) as an indicator of cellular oxidative DNA base damage are biomarkers of radiation-induced cytogenetic damage. The course of both biomarkers was studied in patients with different types of chronic synovitis undergoing radiation synovectomy with very short-lived 165Dy-ferric-hydroxide (DFH). METHODS: Radiation synovectomy of the knee was performed in 13 men and 12 women (mean age, 44+/-15 y) using a mean activity of 9.48+/-1.65 GBq 165Dy-DFH in 27 consecutive treatments. MN frequency in lymphocytes and urinary excretion of 8OHdG, measured by high-performance liquid chromatography, were assessed before and 4 (MN only) and 20 h after radiation synovectomy. RESULTS: Urinary excretion of 8OHdG in patients (in micromol/mol creatinine; pretreatment mean, 3.1+/-3.4; median, 2.27) was not significantly different from that in healthy volunteers (mean, 2.0+/-1.2; median, 1.87) and not altered by radiation synovectomy (post treatment mean, 2.5+/-1.5; median, 2.04, NS). An increase in 8OHdG levels after radiation synovectomy of more than 1 SD was found in only 1 patient, who experienced leakage to the lymph nodes but who already had elevated urinary 8OHdG levels before treatment. The frequency of MN/500 binucleated cells (BNCs) was slightly lower in patients (pretreatment mean, 4.3+/-2.6; median, 4.25) than in healthy volunteers (mean, 5.4+/-2.3; median, 5.3) and did not significantly change after therapy, either (4-h post-treatment mean, 3.9+/-2.1, median, 3.8; 20 h post-treatment mean, 4.1+/-2, median 3.8 MN/500 BNC). In 22 of 27 treatments, no leakage to nontarget organs could be monitored, whereas leakage to the local lymph nodes and the liver was detected after 5 treatments. CONCLUSION: Radiation synovectomy using 165Dy-DFH causes no significant radiation burden to most patients as indicated by the absence of adverse changes in levels of biomarkers of cytogenetic damage and a low incidence of leakage. These data suggest that the risk of malignancy may not be elevated. PMID- 10688108 TI - Sjogren's syndrome: comparison of assessments with quantitative salivary gland scintigraphy and contrast sialography. AB - This study compared the quantitative parameters of salivary gland scintigraphy and the sialographic stages in patients with Sjogren's syndrome. METHODS: One hundred sixteen patients suspected of having Sjogren's syndrome were examined with salivary gland scintigraphy and contrast sialography. When contrast sialography was used as the gold standard, Sjogren's syndrome was diagnosed in 50 of these 116 patients; Sjogren's syndrome was not seen in the other 66 patients. After injection of 370 MBq 99mTc-sodium pertechnetate, dynamic salivary gland scintigraphy with lemon juice stimulation was performed for 50 min. Functional parameters for the parotid and submandibular glands were calculated, and scintigraphic and sialographic results were compared. RESULTS: With the progression of sialographic stages from 0 to 4, the quantity of tracer accumulation decreased in the submandibular gland (P < 0.0001), and the quantity of tracer secretion decreased in the parotid gland (P < 0.0001). The sialographic stage in patients with Sjogren's syndrome was correlated with these scintigraphic parameters (P < 0.0001): sialographic stage = 3.243 - 0.337 x (submandibular gland uptake ratio) - 0.026 x (parotid gland maximum secretion). CONCLUSION: The decreased accumulation in the submandibular gland and the decreased secretion in the parotid gland were highly sensitive indicators of salivary gland disease in Sjogren's syndrome. The sialographic stage was correlated with these scintigraphic parameters. PMID- 10688109 TI - Fewer women than men have positive SPECT and PET cardiac findings among patients with no history of heart disease. AB - A lower detection rate of coronary artery disease (CAD) has been reported for SPECT imaging in women, despite the fact that similar numbers of women and men die each year of heart disease. Ruling out instrumentation as a possible source of this low detection rate for CAD in women could be important in determining the root cause of this difference. METHODS: Patients were referred by cardiologists and randomized to PET or SPECT by the imaging center. A total of 210 patients (106 women, 104 men) were enrolled in this study, with 105 imaged by dual-isotope SPECT and 105 imaged by 82Rb PET. Rest/stress scanning was performed using dipyridamole. The effects of sex, prior history of CAD, and instrumentation on the detection of positive scans were determined using multiple logistic regression analysis with positive scans as the endpoint. RESULTS: For the total study population, sex and prior history of CAD are significantly associated with positive scans, whereas imaging modality and age are not. There was no significant interaction between sex and prior history of CAD. Men have 4.1-fold greater odds of having a positive nuclear scan than women, and patients with prior history of CAD have 5.2-fold greater odds of a positive scan after controlling for the confounding effects of age and imaging modality. In the subgroup of patients with no prior history of heart disease, men have 3.9-fold greater odds of a positive scan than women, and the odds ratio of a positive scan is 2.5-fold greater for PET than for SPECT. There was no statistical difference in the number of positive scans by SPECT or PET, or positive scans by sex in patients with documented history of CAD. CONCLUSION: Fewer women than men have positive nuclear cardiology scans by both PET and SPECT, despite similar symptoms. Instrumentation characteristics alone do not account for this sex-based difference and suggest the possibility that early CAD may present differently in women than in men. PMID- 10688110 TI - When is hilar uptake of 67Ga-citrate indicative of residual disease after CHOP chemotherapy? AB - The purpose of this study was to evaluate the prevalence and characterize the patterns of hilar uptake (HU) on 67Ga-citrate imaging after cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy regimens for non Hodgkin's lymphoma (NHL), to differentiate hilar lymphoma (HL) from HU of benign etiology. METHODS: A total of 930 studies (698 planar, 232 thoracic SPECT) was reviewed retrospectively in 100 NHL patients (29 low-grade, 60 intermediate grade, and 11 high-grade) treated with CHOP and followed up longitudinally with serial gallium studies (planar: median, 7; range, 3-16 studies in 100 patients; SPECT: median, 1; range, 0-11 studies in 72 patients) over a median duration of 36 mo (range, 6-112 mo) from diagnosis. Clinical outcome and size changes over time on correlative CT and/or radiographs were used to evaluate benign versus malignant changes within the hila. RESULTS: HU after CHOP was present in 79% of patients (90% confidence interval [CI], 71%-85%), with 33% showing HU on SPECT alone. Once present, HU persisted for a median of 27 mo (range, 2-84 mo) from onset. The prevalence of HU and HL at various time points was as follows: baseline HU, 52% with HL 60%; mid-CHOP HU, 59% with HL2%; post-CHOP HU, 52% with HL6%; follow-up HU, 76% with HL 9%. HU of benign etiology was not significantly correlated with CHOP dosage. HU was symmetric in 90% of patients (90% CI, 82% 95%) and less intense than the original disease in 89% of patients (90% CI, 80% 95%), and these features were highly predictive of benign etiology (negative predictive value [NPV], 98.6% if symmetric; NPV, 96.5% if less than original disease; NPV, 100% if both present). Asymmetric HU equal in intensity to the original disease, however, was highly predictive of HL (positive predictive value [PPV], 87.5% if asymmetric; PPV, 85.7% if equal to original disease; PPV, 100% if both present). CONCLUSION: HU after CHOP is common (overall incidence, 79%), often seen only on SPECT, and most likely of benign etiology when symmetric and less intense than the original disease. Asymmetric HU that equals the intensity of the original disease, however, is a possible indicator for HL. PMID- 10688111 TI - PET imaging of adrenal cortical tumors with the 11beta-hydroxylase tracer 11C metomidate. AB - The purpose of the study was to evaluate PET with the tracer 11C-metomidate as a method to identify adrenal cortical lesions. METHODS: PET with 11C-metomidate was performed in 15 patients with unilateral adrenal mass confirmed by CT. All patients subsequently underwent surgery, except 2 who underwent biopsy only. The lesions were histopathologically examined and diagnosed as adrenal cortical adenoma (n = 6; 3 nonfunctioning), adrenocortical carcinoma (n = 2), and nodular hyperplasia (n = 1). The remaining were noncortical lesions, including 1 pheochromocytoma, 1 myelolipoma, 2 adrenal cysts, and 2 metastases. RESULTS: All cortical lesions were easily identified because of exceedingly high uptake of 11C metomidate, whereas the noncortical lesions showed very low uptake. High uptake was also seen in normal adrenal glands and in the stomach. The uptake was intermediate in the liver and low in other abdominal organs. Images obtained immediately after tracer injection displayed high uptake in the renal cortex and spleen. The tracer uptake in the cortical lesions increased throughout the examination. For quantitative evaluation of tracer binding in individual lesions, a model with the splenic radioactivity concentration assigned to represent nonspecific uptake was applied. Values derived with this method, however, did show the same specificity as the simpler standardized uptake value concept, with similar difference observed for cortical versus noncortical lesions. CONCLUSION: PET with 11C-metomidate has the potential to be an attractive method for the characterization of adrenal masses with the ability to discriminate lesions of adrenal cortical origin from noncortical lesions. PMID- 10688113 TI - Can 11C-methionine play a role in lung cancer staging? PMID- 10688112 TI - Usefulness of PET with 11C-methionine for the detection of hilar and mediastinal lymph node metastasis in lung cancer. AB - We retrospectively evaluated the usefulness of PET with 11C-methionine (methionine PET) for the diagnosis of lymph node metastases in patients with lung cancer. METHODS: Methionine PET and CT were performed before surgical intervention in 41 patients with primary lung cancer. We evaluated individual lymph nodes by methionine PET along with corresponding CT images. The 11C methionine accumulation of lymph nodes was assessed semiquantitatively by analysis of the tumor-to-muscle ratio (TMR) and was compared with CT and histological diagnoses. RESULTS: A total of 126 lymph nodes, 36 of which were histologically diagnosed as metastatic, were assessed by CT and methionine PET. The TMR in metastatic lymph nodes (n = 36) was 5.15+/-1.69, whereas that of nonmetastatic lymph nodes (n = 90) was 2.91+/-0.76; this difference was significant (P < 0.0001). The most adequate TMR cutoff value for diagnosis of metastasis based on the results of receiver operating characteristic curve analysis was 4.1. The positive and negative predictive values, sensitivity, specificity, and accuracy of methionine PET were 79.5%, 94.3%, 86.1%, 91.1%, and 89.7%, respectively, and were superior to those of CT (57.6%, P = 0.04; 81.7%, P = 0.008; 52.8%, P = 0.002; 84.4%, NS; and 75.4%, P = 0.002, respectively). All positive nodes that were shown to be true-positive by CT, and 12 of 17 false negatives on CT were correctly diagnosed by PET. Ten of 14 lymph nodes that were false-positive on CT were also correctly diagnosed by PET. CONCLUSION: Methionine PET appears to be superior to CT for the diagnosis of lymph node metastasis in lung cancer patients. The high negative predictive value of methionine PET suggests that cases in which lymph nodes are enlarged on CT with negative PET analysis may be diagnosed as negative for metastasis. PMID- 10688114 TI - Iodinated free fatty acid and 201T1 uptake in chronically hypoperfused myocardium: histologic correlation study. AB - 123I-15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) is a tracer for the evaluation of ischemic heart disease. The purpose of this study was to assess the relationship between 1231-BMIPP uptake and myocardial fibrosis. METHODS: Fifteen patients who underwent cardiac surgery were examined by imaging with 201TI and 123I-BMIPP, and histologic specimens were taken during surgery. The relative uptake of 201TI (%TI) and that of 123I-BMIPP (%BMIPP) were calculated. The percentage of fibrosis (%fibrosis) was analyzed with the specimen. RESULTS: %TI correlated strongly with %fibrosis (r = -0.94; P < 0.001). %BMIPP also correlated significantly with %fibrosis (r = -0.88; P < 0.001), but the change in %BMIPP looked biphasic. In the category of only mild fibrosis, %BMIPP showed a steep decrease. 123I-BMIPP-201TI mismatch was found only for fibrosis <20%. CONCLUSION: 123I-BMIPP gave specific information about metabolic changes that occurred in ischemic myocardium without severe fibrotic changes. PMID- 10688115 TI - Internal radionuclide radiation dosimetry: a review of basic concepts and recent developments. AB - Internal dosimetry deals with the determination of the amount and the spatial and temporal distribution of radiation energy deposited in tissue by radionuclides within the body. Nuclear medicine has been largely a diagnostic specialty, and model-derived average organ dose estimates for risk assessment, the traditional application of the MIRD schema, have proven entirely adequate. However, to the extent that specific patients deviate kinetically and anatomically from the model used, such dose estimates will be inaccurate. With the increasing therapeutic application of internal radionuclides and the need for greater accuracy, radiation dosimetry in nuclear medicine is evolving from population- and organ average to patient- and position-specific dose estimation. Beginning with the relevant quantities and units, this article reviews the historical methods and newly developed concepts and techniques to characterize radionuclide radiation doses. The latter include the 3 principal approaches to the calculation of macroscopic nonuniform dose distributions: dose point-kernel convolution, Monte Carlo simulation, and voxel S factors. Radiation dosimetry in "sensitive" populations, including pregnant women, nursing mothers, and children, also will be reviewed. PMID- 10688116 TI - Performance of a 62Zn/62Cu generator in clinical trials of PET perfusion agent 62Cu-PTSM. AB - The 62Zn/62Cu PET generator can be inexpensively produced and distributed from a single production site operating under typical good manufacturing practice guidelines. It therefore has the potential to greatly facilitate development of clinically practical PET. We report generator performance in a study in which 62Cu-pyruvaldehyde-bis(n4-methylthiosemicarbazone (PTSM) myocardial perfusion imaging is compared with 99mTc-sestamibi in the diagnosis of coronary artery disease. The 62Zn/62Cu generator is an improved version of a previously reported system that employs automated synthesis of 62Cu-PTSM. With this approach, the cumbersome step of 18C purification has been eliminated. METHODS: The 62Zn (9.3 h half-life) parent isotope is prepared by proton bombardment of natural copper at 33 MeV. A typical target irradiated with 37.5 microA/h is delivered by 12:00 PM on the day it is to be processed. Purified 62Zn obtained from the target is loaded onto the generator column in 2 mol/L HCl. The generator is eluted using an internal three-channel peristaltic pump, which delivers 2.25 mL eluant (1.8 mol/L NaCl, 0.2 mol/L HCl) through the generator column to elute the 62Cu in 40 s. The same pump simultaneously pumps an equal volume of buffer (0.4 mol/L NaOAc) and 1 mL ligand solution (2 ppm PTSM, 2% EtOH) passing it through a septum into a 35-cc syringe preloaded with 28 mL sterile water. This solution is thoroughly mixed by agitation of the syringe and injected as a bolus through a 0.2 microm filter. The generator is eluted twice before shipping, providing quality assurance samples, and shipped to the clinical site by overnight delivery. Complete quality assurance testing is performed the evening before the generator reaches the clinical site. RESULTS: A total of 34 generators have been produced and shipped to 2 clinical sites for a phase III Food and Drug Administration study. The load activity on the generators at 8:00 AM the day of clinical use was 1.7+/-0.2 GBq (46.7+/-5.6 mCi), and yield was 72%+/-16%. Breakthrough of 62Zn was undetectable by high-purity germanium spectroscopy for all units. Radiochemical purity was 95.4%+/-2.4%. Volume delivered, pH, sterility, and bacterial endotoxin tests yielded passing results on all generators. The entire process of generator production, from target receipt to generator shipment, took less than 6 h and cost approximately $1000, including shipping charges and cyclotron cost. A total of 68 patients were injected with 2 62Cu-PTSM doses, with a mean injected activity of 0.8+/-0.2 GBq (20.5+/-5.3 mCi) with no adverse side effects. CONCLUSION: Results of this work confirm that the 62Zn/62Cu generator is an easily produced, transportable, and inexpensive source of PET radiopharmaceuticals, which can expand the field of clinical PET imaging by providing radiopharmaceuticals to sites not associated with cyclotrons. PMID- 10688117 TI - The potential role of generator-produced radiopharmaceuticals in clinical PET. PMID- 10688118 TI - Effects of insulinlike growth factor binding proteins on insulinlike growth factor-I biodistribution in tumor-bearing nude mice. AB - This study evaluated the biodistribution and tumor targeting ability of radiolabeled insulinlike growth factor (IGF)-I. Because IGF binding proteins (IGFBPs) play a critical role in modulating IGF activity, the binding properties of 125I-labeled IGF-I to IGFBPs were investigated in vitro and in vivo. Because a large amount of the IGF-I was catabolized in vivo, we also studied the catabolism of IGF-I by tumor cells in vitro. METHODS: 125I-labeled-IGF-I was prepared using the chloramine T method. The biodistribution of 125I-labeled-IGF-I in tumor bearing nude mice was compared between groups injected with 125I-labeled IGF-I alone or coinjected with unlabeled peptide. In vitro and in vivo chromatography studies were performed to evaluate the binding profile to IGFBPs and the degree of catabolites in serum as well as urine. RESULTS: Data indicated that the binding of radiolabeled IGF-I to IGFBPs in vitro was dose dependent. However, there was a difference in complex formation between the serum and the heparinized plasma. In heparinized plasma, the radioactivity shifted from a 30- to 50-kDa complex to a 150-kDa complex and to a free ligand, because the binding of heparin with IGFBPs decreased its affinity for IGF-I. In plasma prepared with acid citrate dextrose a binding pattern identical to that of serum was observed. Moreover, there was a binding difference between mouse and rat. The 125I-labeled IGF-I catabolized very quickly when incubated at 37 degrees C but not at all at 4 degrees C. In tumor-bearing nude mice, the uptake of radioactivity in normal tissues decreased quickly, particularly in the kidneys. In mice coinjected with unlabeled carrier, the radioactivity in most normal tissues was lower and the tumor uptake higher than in the mice without carrier. CONCLUSION: These data confirm that 125I-labeled IGF-I is avidly bound to IGFBPs, both in vitro and in vivo. By partially saturating this binding with unlabeled peptides, a favorable biodistribution was achieved, including faster clearance from normal tissue and higher tumor uptake, which resulted in better tumor-to-nontumor ratios. Nevertheless, the rapid catabolism and release of the radiolabel from tumor tissue result in a suboptimal targeting agent. PMID- 10688119 TI - Noninvasive detection of tumor hypoxia using the 2-nitroimidazole [18F]EF1. AB - The noninvasive assessment of tumor hypoxia in vivo is under active investigation because hypoxia has been shown to be an important prognostic factor for therapy resistance. Various nuclear medicine imaging modalities are being used, including PET imaging of 18F-containing compounds. In this study, we report the development of 18F-labeled EF1 for noninvasive imaging of hypoxia. EF1 is a 3-monofluoro analog of the well-characterized hypoxia marker EF5, 2(2-nitro-1H-imidazol-1-yl) N-(2,2,3,3,3-pentafluoropropyl)acetami de, which has been used to detect hypoxia in tumor and nontumor systems using immunohistochemical methods. METHODS: We have studied 2 rat tumor types: the hypoxic Morris 7777 (Q7) hepatoma and the oxic 9LF glioma tumor, each grown in subcutaneous sites. PET studies were performed using a pharmacological dose of nonradioactive carrier in addition to [18F]EF1 to optimize and assess drug biodistribution. After PET imaging of the tumor-bearing rats, tissues were obtained for gamma-counting of the 18F in various tissues and immunohistochemical detection of intracellular drug adducts in tumors. In one pair of tumors, Eppendorf needle electrode studies were performed. RESULTS: [18F]EF1 was excreted dominantly through the urinary tract. The tumor-to-muscle (T/M) ratio of [18F]EF1 in the Q7 tumors was 2.7 and 2.4 based on PET studies and 2.1, 2.5, and 3.0 based on gamma-counting of the tissues (n = 3). In contrast, the T/M ratio of [18F]EF1 in the 9LF glioma tumor was 0.8 and 0.5 based on PET studies and 1.0, 1.2, and 1.4 based on gamma-counting of the tissues (n = 3). Immunohistochemical analysis of drug adducts for the two tumor types agreed with the radioactivity analysis. In the Q7 tumor, substantial heterogeneous binding was observed throughout the tumor, whereas in the 9LF tumor minimal binding was found. CONCLUSION: [18F]EF1 is an excellent radiotracer for noninvasive imaging of tumor hypoxia. PMID- 10688120 TI - CaNa2EDTA for improvement of radioimmunodetection and radioimmunotherapy with 111In and 90Y-DTPA-anti-CEA MAbs in nude mice bearing human colorectal cancer. AB - 111In and 90Y, dissociated from 111In-labeled-monoclonal antibody (MAb) and 90Y labeled MAb, may cause deterioration of the image quality in radioimmunodetection (RID) and undesirable irradiation of nontargeted tissue in radioimmunotherapy (RIT), respectively. The aim of this study was to investigate any improvement in RID and RIT with 111In-MAb and 90Y-MAb by pre- and postadministration of calcium disodium ethylenetriaminetetraacetic acid (CaNa2EDTA). METHODS: Murine MAb F33 104 against carcinoembryonic antigen (CEA) was labeled with 111In or 90Y by the diethylenetriamine pentaacetic (DTPA)-anhydride method. The influence of CaNa2EDTA on loss of radioactivity from 111In-MAb or 90Y-MAb in serum was investigated in vitro. The effects of CaNa2EDTA, administered before and after 111In-MAb or 90Y-MAb, on the biodistribution of radioactive isotopes in nude mice bearing human colon adenocarcinoma LS 180 tumor expressing CEA, or human pulmonary carcinoma PC 9 tumor expressing no CEA, were then examined. As a control, 0.9% NaCl was used in both the in vitro and in vivo studies. RESULTS: CaNa2EDTA did not cause any decrease in levels of radioactivity of radiolabeled MAbs. Pre- and post-treatment with CaNa2EDTA reduced radioctivity in both specific and nonspecific tumors at 72 h after 111In-MAb injection resulting in an increase of the specific tumor-to-nonspecific tumor radioactivity ratio. The levels of hepatic and renal radioactivity were also subsequently decreased by CaNa2EDTA. On the other hand, CaNa2EDTA pre- and post-treatment reduced levels of bony, hepatic, and renal radioactivity at 24, 72, and 72 h, respectively, after 90Y-MAb injection, although it had no effect on tumor radioactivity. CONCLUSION: Pre- and post-treatment with CaNa2EDTA would be of great use in humans who undergo RID or RIT with 111In-MAb and 90Y-MAb accompanied by disassociation of the labeled radionuclides. PMID- 10688121 TI - 11C-labeled KF18446: a potential central nervous system adenosine A2a receptor ligand. AB - To develop PET ligands for mapping central nervous system (CNS) adenosine A2a receptors that are localized in the striatum and are coupled with dopamine receptors, 3 11C-labeled xanthine-type adenosine A2a antagonists, [11C]KF18446 ([7-methyl-11C]-(E)-8-(3,4,5-trimethoxystyryl)-1,3,7-trimethylxanthin e), [11C]KF19631 ([7-methyl-11C]-(E)-1,3-diallyl-7-methyl-8-(3,4,5 trimethoxystyryl)xanth ine), and [11C]CSC ([7-methyl-11C]-8 chlorostyrylcaffeine), were compared with [11C]KF17837 ([7-methyl-11C]-(E)-8-(3,4 dimethoxystyryl)-1,3-dipropyl-7-methylx anthine). METHODS: The regional brain uptake of the tracers, the effect of the coinjected adenosine antagonists on the uptake, and the metabolism were studied in mice. In rats, the regional brain uptake of the tracers was visualized by ex vivo autoradiography (ARG). The A2a receptor binding of antagonist 1 was also measured by in vitro ARG. Imaging of the monkey brain was performed with PET with antagonist 1. RESULTS: In mice, the highest striatal uptake was found for antagonist 1 followed by antagonists 2 and 4. The uptake was inhibited by each of 3 KF compounds and by CSC, but not by an A1 antagonist KF15372. Another selective nonxanthine-type A2a antagonist SCH 58261 significantly decreased the striatal uptake of only antagonist 1, the labeled metabolites of which were less than 20% in the plasma 30 min postinjection, but were negligible in the brain tissue. In ex vivo ARG, antagonist 1 showed the highest striatal uptake and the highest uptake ratio of the striatum to the other brain regions. A high and selective binding of antagonist 1 to the striatum was also confirmed by in vitro ARG. PET with antagonist 1 visualized adenosine A2a receptors in the monkey striatum. CONCLUSION: These results indicate that antagonist 1 ([11C]KF18446) is the most suitable PET ligand for mapping adenosine A2a receptors in the CNS. PMID- 10688122 TI - Single amino acid substitution in the Fc region of chimeric TNT-3 antibody accelerates clearance and improves immunoscintigraphy of solid tumors. AB - Recent studies in antibody catabolism have identified residues at the CH2-CH3 interface of the IgG heavy chain critical for serum persistence of immunoglobulins. Amino acid substitutions in the Fc region of murine IgG1 were shown to drastically accelerate antibody clearance in mice. Our laboratory has previously described a human-mouse chimeric TNT-3 (chTNT-3) monoclonal antibody directed against a universal nuclear antigen that has potential for the radioimmunotherapy of many solid tumors. In the current study, we engineered a chTNT-3 mutant containing a single amino acid substitution, to determine whether a more rapid clearance profile would make the antibody suitable for diagnostic imaging. METHODS: A single amino acid substitution in the CH2 domain of the human gamma1 constant region was made by polymerase chain reaction mutagenesis. High level expression was achieved using the Glutamine Synthetase Gene Amplification System, and the chTNT-3 mutant was purified by protein A affinity and ion exchange chromatography. A radioimmunoassay was performed to examine antigen binding, and in vivo studies were undertaken to evaluate clearance and tumor targeting in human tumor xenograft models. RESULTS: The chTNT-3 mutant retained the high affinity of chTNT-3, with a binding constant of 1.5 x 10(-9) mol/L. The mutant was eliminated rapidly from BALB/c mice, with a beta-phase half-life of 33.8 h, compared to 134.2 h for chTNT-3. Moreover, biodistribution studies in human colon tumor-bearing nude mice reflected this accelerated clearance. Tumor levels of the mutant were, respectively, 65%, 39%, and 36% of the tumor levels achieved with the parental chTNT-3 6, 12, and 24 h postinjection. The rapid clearance of the chTNT-3 mutant from the blood resulted in higher tumor-to-normal organ ratios for many normal tissues. Imaging of tumor-bearing mice with 99mTc labeled chTNT-3 mutant demonstrated early visualization of tumors in 3 different solid tumor xenograft models. CONCLUSION: The accelerated clearance produced by a single amino acid substitution in the Fc region of chTNT-3 leads to improved imaging in tumor-bearing mice. These studies suggest that a rapidly clearing antibody generated by this approach may be useful for the immunoscintigraphy of human tumors. PMID- 10688123 TI - Inflammation and infection imaging with a 99mTc-neutrophil elastase inhibitor in monkeys. AB - A radiolabeled human neutrophil elastase inhibitor (EPI-HNE-2) may represent an improved nuclear medicine imaging agent for inflammation and infection. This peptide displays rapid pharmacokinetics due to its low molecular weight and localizes specifically on neutrophil elastase released in inflammatory sites by activated neutrophils. METHODS: In this investigation, the peptide was radiolabeled with 99mTc using N-hydroxysuccinimidyl S acetylmercaptoacetyltriglycline (NHS-MAG3) as a bifunctional chelator and was administered on 18 occasions to 5 rhesus monkeys with inflammation/infection. RESULTS: Plasma clearance was rapid, with liver and kidneys representing the major organs of accumulation. No evidence of toxicity, dosage effects, or circulating antiMAG3-EPI-HNE-2 antibodies was observed. Specificity of localization was established using radiolabeled bovine pancreatic trypsin inhibitor (a non-hNE-binding peptide of similar size) as a nonspecific negative control peptide and by predosing with unlabeled EPI-HNE-2 to block receptor sites before the administration of radiolabeled EPI-HNE-2. The ability of radiolabeled EPI-HNE-2 to image inflammation/infection was evaluated in 12 studies in monkeys receiving only radiolabeled EPI-HNE-2 and with lesions in the arm, shoulder, or lower back. Positive images were obtained in all studies, uptake was apparent almost immediately, and images were still positive 24 h later. As a positive control, animals also received nonspecific IgG antibody radiolabeled with 99mTc either directly or by NHS-MAG3. Compared with labeled antibody, plasma clearance of 99mTc was faster with labeled EPI-HNE-2 and accumulation in liver and heart was lower. Uptake of radioactivity in the inflammation was higher during the first hour with EPI-HNE-2 versus antibody but lower thereafter. CONCLUSION: When radiolabeled with 99mTc, EPI-HNE-2 localized specifically in inflammations in a monkey model and provided early images of diagnostic quality. PMID- 10688124 TI - Nuclear medicine image registration by spatially noncoherent interferometry. AB - This article introduces a technique for obtaining high-resolution body contour data in the same coordinate frame as that of a rotating gamma camera, using a miniature range finder, the conoscope, mounted on the camera gantry. One potential application of the technique is accurate coregistration in longitudinal brain SPECT studies, using the face of the patient (or "mask"), instead of SPECT slices, to coregister subsequent acquisitions involving the brain. METHODS: Conoscopic holography is an interferometry technique that relies on spatially incoherent light interference in birefringent crystals. In this study, the conoscope was used to measure the absolute distance (Z) between a light source reflected from the skin and its observation plane. This light was emitted by a 0.2-mW laser diode. A scanning system was used to image the face during SPECT acquisition. The system consisted of a motor-driven mirror (Y axis) and the gamma camera gantry (1 profile was obtained for each rotation step, X axis). The system was calibrated to place the conoscopic measurements and SPECT slices in the same coordinate frame. RESULTS: Through a simple and robust calibration of the system, the SE for measurements performed on geometric shapes was less than 2 mm, i.e., less than the actual pixel size of the SPECT data. Biometric measurements of an anthropomorphic brain phantom were within 3%-5% of actual values. The mask data were used to register images of a brain phantom and of a volunteer's brain, respectively. The rigid transformation that allowed the merging of masks by visual inspection was applied to the 2 sets of SPECT slices to perform the fusion of the data. CONCLUSION: At the cost of an additional low-cost setup integrated into the gamma-camera gantry, real-time data about the surface of the head were obtained. As in all other surface-based techniques (as opposed to volume-based techniques), this method allows the match of data independently from the dataset of interest and facilitates further registration of data from any other source. The main advantage of this technique compared with other optically based methods is the robustness of the calibration procedure and the compactness of the sensor as a result of the colinearity of the projected beam and the reflected (diffused) beams of the conoscope. Taking into account the experimental nature of this preliminary work, significant improvements in the accuracy and speed of measurements (up to 1000 points/s) are expected. PMID- 10688125 TI - National Electrical Manufacturers Association recommendations for implementing SPECT instrumentation quality control. PMID- 10688126 TI - Effect of obesity on red cell mass. PMID- 10688127 TI - Mammography and 99mTc-MIBI scintimammography in suspected breast cancer. PMID- 10688128 TI - Evolution of genes and taxa: a primer. AB - The rapidly growing fields of molecular evolution and systematics have much to offer to molecular biology, but like any field have their own repertoire of terms and concepts. Homology, for example, is a central theme in evolutionary biology whose definition is complex and often controversial. Homology extends to multigene families, where the distinction between orthology and paralogy is key. Nucleotide sequence alignment is also a homology issue, and is a key stage in any evolutionary analysis of sequence data. Models based on our understanding of the processes of nucleotide substitution are used both in the estimation of the number of evolutionary changes between aligned sequences and in phylogeny reconstruction from sequence data. The three common methods of phylogeny reconstruction--parsimony, distance and maximum likelihood--differ in their use of these models. All three face similar problems in finding optimal--and reliable -solutions among the vast number of possible trees. Moreover, even optimal trees for a given gene may not reflect the relationships of the organisms from which the gene was sampled. Knowledge of how genes evolve and at what rate is critical for understanding gene function across species or within gene families. The Neutral Theory of Molecular Evolution serves as the null model of molecular evolution and plays a central role in data analysis. Three areas in which the Neutral Theory plays a vital role are: interpreting ratios of nonsynonymous to synonymous nucleotide substitutions, assessing the reliability of molecular clocks, and providing a foundation for molecular population genetics. PMID- 10688129 TI - Examining rates and patterns of nucleotide substitution in plants. AB - Driven by rapid improvements in affordable computing power and by the even faster accumulation of genomic data, the statistical analysis of molecular sequence data has become an active area of interdisciplinary research. Maximum likelihood methods have become mainstream because of their desirable properties and, more importantly, their potential for providing statistically sound solutions in complex data analysis settings. In this chapter, a review of recent literature focusing on rates and patterns of nucleotide substitution rates in the nuclear, chloroplast, and mitochondrial genomes of plants demonstrates the power and flexibility of these new methods. The emerging picture of the nucleotide substitution process in plants is a complex one. Evolutionary rates are seen to be quite variable, both among genes and among plant lineages. However, there are hints, particularly in the chloroplast, that individual factors can have important effects on many genes simultaneously. PMID- 10688130 TI - Contributions of plant molecular systematics to studies of molecular evolution. AB - Dobzhansky stated that nothing in biology makes sense except in the light of evolution. A close corollary, and the central theme of this paper, is that everything makes a lot more sense in the light of phylogeny. Systematics is in the midst of a renaissance, heralded by the widespread application of new analytical approaches and the introduction of molecular techniques. Molecular phylogenetic analyses are now commonplace, and they have provided unparalleled insights into relationships at all levels of plant phylogeny. At deep levels, molecular studies have revealed that charophyte green algae are the closest relatives of the land plants and suggested that liverworts are sister to all other extant land plants. Other studies have suggested that lycopods are sister to all other vascular plants and clarified relationships among the ferns. The impact of molecular phylogenetics on the angiosperms has been particularly dramatic--some of the largest phylogenetic analyses yet conducted have involved the angiosperms. Inferences from three genes (rbcL, atpB, 18S rDNA) agree in the major features of angiosperm phylogeny and have resulted in a reclassification of the angiosperms. This ordinal-level reclassification is perhaps the most dramatic and important change in higher-level angiosperm taxonomy in the past 200 years. At lower taxonomic levels, phylogenetic analyses have revealed the closest relatives of many crops and 'model organisms' for studies of molecular genetics, concomitantly pointing to possible relatives for use in comparative studies and plant breeding. Furthermore, phylogenetic information has contributed to new perspectives on the evolution of polyploid genomes. The phylogenetic trees now available at all levels of the taxonomic hierarchy for angiosperms and other green plants should play a pivotal role in comparative studies in diverse fields from ecology to molecular evolution and comparative genetics. PMID- 10688131 TI - Molecular evolution of the chalcone synthase multigene family in the morning glory genome. AB - Plant genomes appear to exploit the process of gene duplication as a primary means of acquiring biochemical and developmental flexibility. Thus, for example, most of the enzymatic components of plant secondary metabolism are encoded by small families of genes that originated through duplication over evolutionary time. The dynamics of gene family evolution are well illustrated by the genes that encode chalcone synthase (CHS), the first committed step in flavonoid biosynthesis. We review pertinent facts about CHS evolution in flowering plants with special reference to the morning glory genus, Ipomoea. Our review shows that new CHS genes are recruited recurrently in flowering plant evolution. Rates of nucleotide substitution are frequently accelerated in new duplicate genes, and there is clear evidence for repeated shifts in enzymatic function among duplicate copies of CHS genes. In addition, we present new data on expression patterns of CHS genes as a function of tissue and developmental stage in the common morning glory (I. purpurea). These data show extensive differentiation in gene expression among duplicate copies of CHS genes. We also show that a single mutation which blocks anthocyanin biosynthesis in the floral limb is correlated with a loss of expression of one of the six duplicate CHS genes present in the morning glory genome. This suggests that different duplicate copies of CHS have acquired specialized functional roles over the course of evolution. We conclude that recurrent gene duplication and subsequent differentiation is a major adaptive strategy in plant genome evolution. PMID- 10688132 TI - Myrosinase: gene family evolution and herbivore defense in Brassicaceae. AB - Glucosinolates are a category of secondary products present primarily in species of the order Capparales. When tissue is damaged, for example by herbivory, glucosinolates are degraded in a reaction catalyzed by thioglucosidases, denoted myrosinases, also present in these species. Thereby, toxic compounds such as nitriles, isothiocyanates, epithionitriles and thiocyanates are released. The glucosinolate-myrosinase system is generally believed to be part of the plant's defense against insects, and possibly also against pathogens. In this review, the evolution of the system and its impact on the interaction between plants and insects are discussed. Further, data suggesting additional functions in the defense against pathogens and in sulfur metabolism are reviewed. PMID- 10688133 TI - A short history of MADS-box genes in plants. AB - Evolutionary developmental genetics (evodevotics) is a novel scientific endeavor which assumes that changes in developmental control genes are a major aspect of evolutionary changes in morphology. Understanding the phylogeny of developmental control genes may thus help us to understand the evolution of plant and animal form. The principles of evodevotics are exemplified by outlining the role of MADS box genes in the evolution of plant reproductive structures. In extant eudicotyledonous flowering plants, MADS-box genes act as homeotic selector genes determining floral organ identity and as floral meristem identity genes. By reviewing current knowledge about MADS-box genes in ferns, gymnosperms and different types of angiosperms, we demonstrate that the phylogeny of MADS-box genes was strongly correlated with the origin and evolution of plant reproductive structures such as ovules and flowers. It seems likely, therefore, that changes in MADS-box gene structure, expression and function have been a major cause for innovations in reproductive development during land plant evolution, such as seed, flower and fruit formation. PMID- 10688134 TI - Knots in the family tree: evolutionary relationships and functions of knox homeobox genes. AB - Knotted-like homeobox (knox) genes constitute a gene family in plants. Class I knox genes are expressed in shoot apical meristems, and (with notable exceptions) not in lateral organ primordia. Class II genes have more diverse expression patterns. Loss and gain of function mutations indicate that knox genes are important regulators of meristem function. Gene duplication has contributed to the evolution of families of homeodomain proteins in metazoans. We believe that similar mechanisms have contributed to the diversity of knox gene function in plants. Knox genes may have contributed to the evolution of compound leaves in tomato and could be involved in the evolution of morphological traits in other species. Alterations in cis-regulatory regions in some knox genes correlate with novel patterns of gene expression and distinctive morphologies. Preliminary data from the analysis of class I knox gene expression illustrates the evolution of complex patterns of knox expression is likely to have occurred through loss and gain of domains of gene expression. PMID- 10688135 TI - Evolutionary genetics of self-incompatibility in the Solanaceae. AB - The self-incompatibility (S) gene in flowering plants has long been appreciated as an example of extreme allelic polymorphism maintained by frequency-dependent selection. Recent studies of population samples of S-allele sequences obtained by RT-PCR from five species of Solanaceae now reveal a picture of conspicuous inter specific variation in both S-allele number and age. Explanations for this variation are examined with reference to current theory. We propose that changes in species' effective population size, particularly those associated with the evolution of different life histories, best account for interspecific differences in both the number and average age of S alleles. PMID- 10688136 TI - The evolution of nodulation. AB - In this review we will first describe the different steps leading to nodule formation, and these will be compared with processes of non-symbiotic plant development and growth. In general, aspects of both actinorhizal as well as rhizobial symbiosis are described, but in several cases, the emphasis will be on the Rhizobium-legume symbiosis because more knowledge of this system is available. Subsequently, the phylogeny of nodulating plants is described and a comparison is made between several aspects of legume and actinorhizal nodulation. At the end of this paper the relationship between nodule symbiosis and endomycorrhizal symbiosis is described, and it is discussed to what extent the development of root nodules involves unique properties, or whether processes and genes have been recruited from common plant development and the endomycorrhizal symbiosis. PMID- 10688137 TI - The evolution of disease resistance genes. AB - Several common themes have shaped the evolution of plant disease resistance genes. These include duplication events of progenitor resistance genes and further expansion to create clustered gene families. Variation can arise from both intragenic and intergenic recombination and gene conversion. Recombination has also been implicated in the generation of novel resistance specificities. Resistance gene clusters appear to evolve more rapidly than other regions of the genome. In addition, domains believed to be involved in recognitional specificity, such as the leucine-rich repeat (LRR), are subject to adaptive selection. Transposable elements have been associated with some resistance gene clusters, and may generate further variation at these complexes. PMID- 10688138 TI - Hybridization, introgression, and linkage evolution. AB - Genetic mapping methods provide a unique opportunity to study the interactions of differentiated genes and genomes in a hybrid genetic background. After a brief discussion of theoretical and analytical concerns, we review the application of these methods to a wide range of evolutionary issues. Map-based studies of experimental hybrids indicate that most postzygotic reproductive barriers in plants are polygenic and that the expression of extreme or novel traits in segregating hybrids (transgressive segregation) results from the complementary action of divergent parental alleles. However, genetic studies of hybrid vigor do not concur in their interpretations of the relative roles of dominance, overdominance, and epistasis. Map-based studies of natural hybrids are much rarer, but the few existing studies confirm the polygenic basis of postzygotic barriers and demonstrate the utility of genetic linkage for detecting cryptic introgression. In addition, studies of experimental and natural hybrid lineages provide compelling evidence that homoploid hybrid speciation has occurred in nature, and that it represents a rapid and repeatable mode of speciation. Data further indicate that this mode is facilitated by strong fertility selection and high chromosomal mutation rates. We recommend that future studies of hybrid genomes focus on natural hybrids, not only because of the paucity of data in this area, but also because of the availability of highly recombinant hybrid genotypes in hybrid zones. Of particular value will be studies of long-lived or difficult to-propagate organisms, which previously have not been amenable to genetic study. PMID- 10688141 TI - TB & HIV, applying advances--Chicago 1999 PMID- 10688139 TI - Genome evolution in polyploids. AB - Polyploidy is a prominent process in plants and has been significant in the evolutionary history of vertebrates and other eukaryotes. In plants, interdisciplinary approaches combining phylogenetic and molecular genetic perspectives have enhanced our awareness of the myriad genetic interactions made possible by polyploidy. Here, processes and mechanisms of gene and genome evolution in polyploids are reviewed. Genes duplicated by polyploidy may retain their original or similar function, undergo diversification in protein function or regulation, or one copy may become silenced through mutational or epigenetic means. Duplicated genes also may interact through inter-locus recombination, gene conversion, or concerted evolution. Recent experiments have illuminated important processes in polyploids that operate above the organizational level of duplicated genes. These include inter-genomic chromosomal exchanges, saltational, non Mendelian genomic evolution in nascent polyploids, inter-genomic invasion, and cytonuclear stabilization. Notwithstanding many recent insights, much remains to be learned about many aspects of polyploid evolution, including: the role of transposable elements in structural and regulatory gene evolution; processes and significance of epigenetic silencing; underlying controls of chromosome pairing; mechanisms and functional significance of rapid genome changes; cytonuclear accommodation; and coordination of regulatory factors contributed by two, sometimes divergent progenitor genomes. Continued application of molecular genetic approaches to questions of polyploid genome evolution holds promise for producing lasting insight into processes by which novel genotypes are generated and ultimately into how polyploidy facilitates evolution and adaptation. PMID- 10688142 TI - Molecular determinants of drug resistance in tuberculosis. AB - Rapid detection of drug-resistant tuberculosis (TB) has become increasingly important in the era of pandemic human immunodeficiency virus infection and antibiotic resistance. The identification of the molecular correlates of antibiotic resistance in Mycobacterium tuberculosis have engendered the development of DNA-based assays for the identification of drug-resistant TB. This review summarizes the recent discoveries concerning resistance to isoniazid, rifampin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin and the quinolones. PMID- 10688143 TI - Lung-specific immune response in tuberculosis. PMID- 10688140 TI - Transposable element contributions to plant gene and genome evolution. AB - Transposable elements were first discovered in plants because they can have tremendous effects on genome structure and gene function. Although only a few or no elements may be active within a genome at any time in any individual, the genomic alterations they cause can have major outcomes for a species. All major element types appear to be present in all plant species, but their quantitative and qualitative contributions are enormously variable even between closely related lineages. In some large-genome plants, mobile DNAs make up the majority of the nuclear genome. They can rearrange genomes and alter individual gene structure and regulation through any of the activities they promote: transposition, insertion, excision, chromosome breakage, and ectopic recombination. Many genes may have been assembled or amplified through the action of transposable elements, and it is likely that most plant genes contain legacies of multiple transposable element insertions into promoters. Because chromosomal rearrangements can lead to speciating infertility in heterozygous progeny, transposable elements may be responsible for the rate at which such incompatibility is generated in separated populations. For these reasons, understanding plant gene and genome evolution is only possible if we comprehend the contributions of transposable elements. PMID- 10688144 TI - What we can learn from the Mycobacterium tuberculosis genome sequencing projects. AB - A major milestone in tuberculosis research occurred in June 1998 with the report of the genomic sequence of Mycobacterium tuberculosis H37Rv. The complete determination of the 4411529 base pairs of the M. tuberculosis genome opens avenues for new scientific opportunities in basic science and clinical research on tuberculosis. In this paper we will review the findings presented by the complete genome and discuss the impact that this sequence will have on the areas of comparative genomics, bacterial pathogenesis, and diagnostics development for tuberculosis. Indirect benefits of the complete genome sequence are anticipated in the areas of drug development and vaccine development as future discoveries in bacterial pathogenesis and immunology accrue. PMID- 10688145 TI - Global AIDS 1981-1999: the response. PMID- 10688146 TI - Use of clinical risk assessments in evaluation of nucleic acid amplification tests for HIV/tuberculosis. PMID- 10688147 TI - Applying advances to the clinic and health care delivery: putting the strategies to work. PMID- 10688148 TI - DNA fingerprinting and contact investigation. PMID- 10688149 TI - Community responses to HIV/TB in Canada: a discussion of issues addressed in the HIV/TB Project of the Canadian AIDS Society. PMID- 10688150 TI - Evaluating tuberculosis control programs: strategies, tools and models. PMID- 10688151 TI - Who gains from international tuberculosis collaboration? PMID- 10688152 TI - Applying advances to the developing world for TB and HIV control. PMID- 10688153 TI - The impact of drug resistance on the global tuberculosis epidemic. PMID- 10688154 TI - Tuberculosis preventive therapy for HIV-infected persons in less developed countries. PMID- 10688155 TI - Translating discoveries into control of tuberculosis in HIV-infected persons. PMID- 10688156 TI - Defining the course of brain structural change and plasticity in schizophrenia. AB - Recent evidence from controlled CT and MRI longitudinal studies suggests that some cerebral ventricular enlargement and hemispheric volumetric reductions (e.g. cerebral atrophy) may have a progressive component in patients with schizophrenia. These studies vary in cohort composition, stage of illness examined, duration of follow-up interval, imaging techniques used, and specific brain regions with findings. They also conflict with earlier evidence suggesting that schizophrenia is a neurodevelopmental disorder with brain pathological deviance occurring prior to the illness onset. The newer brain imaging reports may be detecting subtle brain plasticity that results from a continuing cortical disruptive process, may be epi-phenomena caused by scanning and image analysis artifacts or may possibly reflect systemic physiological fluctuations. Future longitudinal studies of subjects at all stages of illness using a variety of new technologies are needed to clarify these findings. PMID- 10688157 TI - A differential neural response to threatening and non-threatening negative facial expressions in paranoid and non-paranoid schizophrenics. AB - Several studies have demonstrated impaired facial expression recognition in schizophrenia. Few have examined the neural basis for this; none have compared the neural correlates of facial expression perception in different schizophrenic patient subgroups. We compared neural responses to facial expressions in 10 right handed schizophrenic patients (five paranoid and five non-paranoid) and five normal volunteers using functional Magnetic Resonance Imaging (fMRI). In three 5 min experiments, subjects viewed alternating 30-s blocks of black-and-white facial expressions of either fear, anger or disgust contrasted with expressions of mild happiness. After scanning, subjects categorised each expression. All patients were less accurate in identifying expressions, and showed less activation to these stimuli than normals. Non-paranoids performed poorly in the identification task and failed to activate neural regions that are normally linked with perception of these stimuli. They categorised disgust as either anger or fear more frequently than paranoids, and demonstrated in response to disgust expressions activation in the amygdala, a region associated with perception of fearful faces. Paranoids were more accurate in recognising expressions, and demonstrated greater activation than non-paranoids to most stimuli. We provide the first evidence for a distinction between two schizophrenic patient subgroups on the basis of recognition of and neural response to different negative facial expressions. PMID- 10688158 TI - Dopamine D2 receptor occupancy by olanzapine or risperidone in young patients with schizophrenia. AB - A crucial characteristic of antipsychotic medication is the occupancy of the dopamine (DA) D2 receptor. We assessed striatal DA D2 receptor occupancy by olanzapine and risperidone in 36 young patients [31 males, 5 females; mean age 21.1 years (16-28)] with first episode schizophrenia, using [123I]iodobenzamide (IBZM) SPECT. The occupancy of DA D2 receptors was not significantly different between olanzapine and risperidone. However, in subgroups of most prescribed doses, DA D2 occupancy was higher in the risperidone 4-mg group (79%) compared to the olanzapine 15-mg group (62%). [123I]IBZM binding ratios decreased with olanzapine dose (r = -0.551; P < 0.01), indicating higher DA D2 receptor occupancy with higher olanzapine dose. Akathisia and positive symptoms were correlated with [123I]IBZM binding ratio (r = -0.442; P < 0.01; and r = -0.360; P < 0.05, respectively). Prolactin (PRL) levels were elevated in the risperidone, but not in the olanzapine group, at comparable D2 receptor occupancy levels. In the olanzapine group, PRL levels were correlated with [123I]IBZM binding ratio (r = -0.551; P < 0.01). In conclusion, both olanzapine and risperidone induce a high striatal D2 receptor occupancy, dependent on dose and group formation. The lower incidence of prolactin elevation with olanzapine, compared to risperidone, may not be attributed to a lower D2 receptor occupancy. PMID- 10688159 TI - Subtype-associated metabolite differences in the temporal lobe in schizophrenia detected by proton magnetic resonance spectroscopy. AB - Brain imaging studies have indicated that the medial temporal lobe functions aberrantly in schizophrenic patients. Both diagnostic subtype and gender may affect functional and morphologic abnormalities in this region. We investigated subtype- and gender-associated differences in metabolites in the left medial temporal lobe in 40 medicated schizophrenic patients by proton magnetic resonance spectroscopy and compared findings with those in 40 healthy control subjects. Peaks corresponding to N-acetylaspartate (NAA), choline-containing compounds (Cho), creatine-phosphocreatine (Cr), and inositol were measured. Schizophrenic patients showed a decrease in the NAA/Cr ratio in the left medial temporal lobe, and patients with the disorganized subtype of illness showed significantly lower NAA/Cr and Cho/Cr ratios than those with paranoid schizophrenia. The NAA/Cr ratio in patients with the undifferentiated subtype also was significantly lower than in the paranoid subtype. No significant associations were observed between metabolite ratios and clinical symptom scores, age at onset of illness, or gender. These findings suggest that patients with the disorganized and undifferentiated subtypes have greater impairments in neuronal integrity or function in the left medial temporal lobe than patients with other subtypes of schizophrenia. PMID- 10688160 TI - NSF boost sends the right message to Congress. PMID- 10688162 TI - CERN claims first experimental creation of quark-gluon plasma. European Laboratory for Particle Physics PMID- 10688161 TI - Think globally, act cautiously. PMID- 10688164 TI - International science council names first female president PMID- 10688163 TI - Austria taken to court for inadequate laws on animal welfare. PMID- 10688165 TI - NASA review leaves projects on launch pad. National Aeronautics & Space Administration PMID- 10688166 TI - Congress gets tough with gene therapy. PMID- 10688167 TI - Scientists reject blame for German genome shortfall. PMID- 10688168 TI - Clinton's farewell gift to US science agencies. PMID- 10688169 TI - Stanford accelerator takes lead in race to quantify CP violation PMID- 10688170 TI - Charm, not tact, aided pioneer in fight for physics PMID- 10688171 TI - Will cell alliance breed bureaucracy and leave contributors out? PMID- 10688172 TI - Proteins suggest form of their own database. PMID- 10688173 TI - Debating controversies can enhance creativity. PMID- 10688174 TI - Time for an aspirin PMID- 10688175 TI - The myth of well-funded German research. PMID- 10688177 TI - Only connect PMID- 10688176 TI - Laying down the law. PMID- 10688179 TI - Dynamics of Jupiter's atmosphere PMID- 10688178 TI - Guilt-by-association goes global. PMID- 10688180 TI - Neuroscience. Images of lost sleep. PMID- 10688181 TI - Enzyme evolution. Design by necessity. PMID- 10688183 TI - Antarctica. Stirring the icy waters PMID- 10688182 TI - Developmental genetics. A Hox by any other name. PMID- 10688184 TI - Cell biology. Which way is up? PMID- 10688185 TI - Flight restores fight in crickets. PMID- 10688186 TI - The ACE gene and muscle performance. PMID- 10688187 TI - A new model for protein stereospecificity. PMID- 10688188 TI - 'Ghost' alleles of the Mauritius kestrel. PMID- 10688189 TI - Directed evolution of new catalytic activity using the alpha/beta-barrel scaffold. AB - In biological systems, enzymes catalyse the efficient synthesis of complex molecules under benign conditions, but widespread industrial use of these biocatalysts depends crucially on the development of new enzymes with useful catalytic functions. The evolution of enzymes in biological systems often involves the acquisition of new catalytic or binding properties by an existing protein scaffold. Here we mimic this strategy using the most common fold in enzymes, the alpha/beta-barrel, as the scaffold. By combining an existing binding site for structural elements of phosphoribosylanthranilate with a catalytic template required for isomerase activity, we are able to evolve phosphoribosylanthranilate isomerase activity from the scaffold of indole-3 glycerol-phosphate synthase. We find that targeting the catalytic template for in vitro mutagenesis and recombination, followed by in vivo selection, results in a new phosphoribosylanthranilate isomerase that has catalytic properties similar to those of the natural enzyme, with an even higher specificity constant. Our demonstration of divergent evolution and the widespread occurrence of the alpha/beta-barrel suggest that this scaffold may be a fold of choice for the directed evolution of new biocatalysts. PMID- 10688190 TI - A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. AB - Two large-scale yeast two-hybrid screens were undertaken to identify protein protein interactions between full-length open reading frames predicted from the Saccharomyces cerevisiae genome sequence. In one approach, we constructed a protein array of about 6,000 yeast transformants, with each transformant expressing one of the open reading frames as a fusion to an activation domain. This array was screened by a simple and automated procedure for 192 yeast proteins, with positive responses identified by their positions in the array. In a second approach, we pooled cells expressing one of about 6,000 activation domain fusions to generate a library. We used a high-throughput screening procedure to screen nearly all of the 6,000 predicted yeast proteins, expressed as Gal4 DNA-binding domain fusion proteins, against the library, and characterized positives by sequence analysis. These approaches resulted in the detection of 957 putative interactions involving 1,004 S. cerevisiae proteins. These data reveal interactions that place functionally unclassified proteins in a biological context, interactions between proteins involved in the same biological function, and interactions that link biological functions together into larger cellular processes. The results of these screens are shown here. PMID- 10688191 TI - Observation of moist convection in Jupiter's atmosphere. Galileo Imaging Team AB - The energy source driving Jupiter's active meteorology is not understood. There are two main candidates: a poorly understood internal heat source and sunlight. Here we report observations of an active storm system possessing both lightning and condensation of water. The storm has a vertical extent of at least 50 km and a length of about 4,000 km. Previous observations of lightning on Jupiter have revealed both its frequency of occurrence and its spatial distribution, but they did not permit analysis of the detailed cloud structure and its dynamics. The present observations reveal the storm (on the day side of the planet) at the same location and within just a few hours of a lightning detection (on the night side). We estimate that the total vertical transport of heat by storms like the one observed here is of the same order as the planet's internal heat source. We therefore conclude that moist convection-similar to large clusters of thunderstorm cells on the Earth-is a dominant factor in converting heat flow into kinetic energy in the jovian atmosphere. PMID- 10688192 TI - Moist convection as an energy source for the large-scale motions in Jupiter's atmosphere. Galileo Imaging Team AB - Jupiter's dominant large-scale weather patterns (dimensions approximately 10,000 km) are zonal jets and long-lived ovals. The jets have been flowing east and west at constant speeds of up to 180 m s(-1) for over 100 years. These jets receive energy from small-scale eddies, which pump eastward momentum into the eastward jets and westward momentum into the westward jets. This momentum transfer was predicted by numerical models before it was observed on Jupiter. The large ovals roll between the jets in an anticyclonic direction-clockwise in the northern hemisphere and counterclockwise in the southern hemisphere--where they regularly assimilate small anticyclonic eddies. But from where the eddies receive their energy has been an open question. Here we argue that the eddies, which ultimately drive both the jets and the ovals, receive their energy from moist convection. This hypothesis is consistent with observations of jovian lightning, which is an indicator of moist convection. It also explains the anticyclonic rotation and poleward drift of the eddies, and suggests patterns of upwelling and downwelling that resemble the patterns of large-scale axisymmetric overturning in the Earth's atmosphere. PMID- 10688193 TI - Structure and bandgap closure in dense hydrogen AB - The possibility that steadily compressed hydrogen might undergo a transition from a proton-paired insulator to a monatomic metal was first suggested in 1935. But experimental realization of metallic hydrogen in solid form has remained elusive, despite studies at pressures as high as 342 GPa. The pairing structure is known to be robust (from the persistence of its associated vibron mode), leading to the suggestion of an alternative route to the metallic state, involving a band overlap transition in which the pairing is preserved. Here we report density functional calculations within the local density approximation that predict a range of densities for hydrogen where a paired or molecular metallic state may be energetically preferred. The transition to this metallic state is naturally associated with the closing of an overall bandgap; but the pressures required to effect the transition are shown to change significantly when the gaps are corrected by approximate inclusion of many-electron effects. The implication is that a complete resolution of the structural and phase problem in dense hydrogen may require methods beyond the local density approximation. PMID- 10688194 TI - Direct measurement of electrical transport through DNA molecules. AB - Attempts to infer DNA electron transfer from fluorescence quenching measurements on DNA strands doped with donor and acceptor molecules have spurred intense debate over the question of whether or not this important biomolecule is able to conduct electrical charges. More recently, first electrical transport measurements on micrometre-long DNA 'ropes', and also on large numbers of DNA molecules in films, have indicated that DNA behaves as a good linear conductor. Here we present measurements of electrical transport through individual 10.4-nm long, double-stranded poly(G)-poly(C) DNA molecules connected to two metal nanoelectrodes, that indicate, by contrast, large-bandgap semiconducting behaviour. We obtain nonlinear current-voltage curves that exhibit a voltage gap at low applied bias. This is observed in air as well as in vacuum down to cryogenic temperatures. The voltage dependence of the differential conductance exhibits a peak structure, which is suggestive of the charge carrier transport being mediated by the molecular energy bands of DNA. PMID- 10688195 TI - Non-destructive determination of local strain with 100-nanometre spatial resolution AB - Structure sizes of approximately 180 nm are now standard in microelectronics, and state-of-the-art fabrication techniques can reduce these to just a few tens of nanometres. But at these length scales, the strain induced at interfaces can locally distort the crystal lattice, which may in turn affect device performance in an unpredictable way. A means of non-destructively characterizing such strain fields with high spatial resolution and sensitivity is therefore highly desirable. One approach is to use Raman spectroscopy, but this is limited by the intrinsic approximately 0.5-microm resolution limit of visible light probes. Techniques based on electron-beam diffraction can achieve the desired nanometre scale resolution. But either they require complex sample preparation procedures (which may alter the original strain field) or they are sensitive to distortional (but not dilational) strain within only the top few tens of nanometres of the sample surface. X-rays, on the other hand, have a much greater penetration depth, but have not hitherto achieved strain analysis with sub-micrometre resolution. Here we describe a magnifying diffraction imaging procedure for X-rays which achieves a spatial resolution of 100nm in one dimension and a sensitivity of 10( 4) for relative lattice variations. We demonstrate the suitability of this procedure for strain analysis by measuring the strain depth profiles beneath oxidized lines on silicon crystals. PMID- 10688196 TI - Air entrapment in coatings by way of a tip-streaming meniscus AB - Entrapment of small air bubbles is a problem for continuous liquid-film coatings processes. The coating of any surface requires that the surrounding air in contact with it be displaced by an advancing liquid interface. Studies of dynamic wetting suggest that if the interface motion is too rapid, the air is not completely removed and it becomes entrained in the coating material. This process, which can lead to undesirable flaws in the form of bubbles, blemishes or voids, limits the speed at which the substrate can be moved in the production of uniform precision coatings. However, the entrapment process is not understood in detail. Here we report an experimental investigation of air entrapment in high speed coating operations. Tip streaming--a phenomenon well known in emulsification technology, involving the ejection of a fine filament from the cusped interface between two immiscible fluids--is shown to be the precursor of air entrainment. We demonstrate that tip-streaming air filaments emanating from the contact zone of a dynamic liquid interface give rise to minute (approximately 10 microm) bubbles. PMID- 10688197 TI - Water exchange between the subglacial Lake Vostok and the overlying ice sheet AB - It has now been known for several years that a 200-km-long lake, called Lake Vostok, lies beneath the ice sheet on which sits Vostok Station in Antarctica. The conditions at the base of the ice sheet above this subglacial lake can provide information about the environment within the lake, including the likelihood that it supports life. Here we present an analysis of the ice-sheet structure from airborne 60-MHz radar studies, which indicates that distinct zones of basal ice loss and accretion occur at the ice-water interface. Subglacial melting and net ice loss occur in the north of the lake and across its 200-km long western margin, whereas about 150 m of ice is gained by subglacial freezing in the south. This indicates that significant quantities of water are exchanged between the base of the ice sheet and the lake waters, which will enrich the lake with gas hydrates, cause sediment deposition and encourage circulation of the lake water. PMID- 10688198 TI - Stable-isotope probing as a tool in microbial ecology. AB - Microorganisms are responsible for driving the biogeochemical cycling of elements on Earth. Despite their importance and vast diversity, the taxonomic identity of the microorganisms involved in any specific process has usually been confined to that small fraction of the microbiota that has been isolated and cultivated. The recent coupling of molecular biological methods with stable-isotope abundance in biomarkers has provided a cultivation-independent means of linking the identity of bacteria with their function in the environment. Here we show that 13C-DNA, produced during the growth of metabolically distinct microbial groups on a 13C enriched carbon source, can be resolved from 12C-DNA by density-gradient centrifugation. DNA isolated from the target group of microorganisms can be characterized taxonomically and functionally by gene probing and sequence analysis. Application of this technique to investigate methanol-utilizing microorganisms in soil demonstrated the involvement of members of two phylogenetically distinct groups of eubacteria; the alpha-proteobacterial and Acidobacterium lineages. Stable-isotope probing thus offers a powerful new technique for identifying microorganisms that are actively involved in specific metabolic processes under conditions which approach those occurring in situ. PMID- 10688199 TI - Ancestral chloroplast genome in Mesostigma viride reveals an early branch of green plant evolution. AB - Sequence comparisons suggest that all living green plants belong to one of two major phyla: Streptophyta (land plants and their closest green algal relatives, the charophytes); and Chlorophyta (the rest of green algae). Because no green algae are known that pre-date the Streptophyta/Chlorophyta split, and also because the earliest diverging green algae show considerable morphological variation, the nature of the unicellular flagellate ancestor of the two green plant phyla is unknown. Here we report that the flagellate Mesostigma viride belongs to the earliest diverging green plant lineage discovered to date. We have sequenced the entire chloroplast DNA (118,360 base pairs) of this green alga and have conducted phylogenetic analyses of sequences derived from this genome. Mesostigma represents a lineage that emerged before the divergence of the Streptophyta and Chlorophyta, a position that is supported by several features of its chloroplast DNA. The structure and gene organization of this genome indicate that chloroplast DNA architecture has been extremely well conserved in the line leading to land plants. PMID- 10688200 TI - Pattern recognition and active vision in chickens. AB - Recognition of objects or environmental landmarks is problematic because appearance can vary widely depending on illumination, viewing distance, angle of view and so on. Storing a separate image or 'template' for every possible view requires vast numbers to be stored and scanned, has a high probability of recognition error and appears not to be the solution adopted by primates. However, some invertebrate template matching systems can achieve recognition by 'active vision' in which the animal's own behaviour is used to achieve a fit between template and object, for example by repeatedly following a set path. Recognition is thus limited to views from the set path but achieved with a minimal number of templates. Here we report the first evidence of similar active vision in a bird, in the form of locomotion and individually distinct head movements that give the eyes a similar series of views on different occasions. The hens' ability to recognize objects is also found to decrease when their normal paths are altered. PMID- 10688201 TI - Altered brain response to verbal learning following sleep deprivation. AB - The effects of sleep deprivation on the neural substrates of cognition are poorly understood. Here we used functional magnetic resonance imaging to measure the effects of 35 hours of sleep deprivation on cerebral activation during verbal learning in normal young volunteers. On the basis of a previous hypothesis, we predicted that the prefrontal cortex (PFC) would be less responsive to cognitive demands following sleep deprivation. Contrary to our expectations, however, the PFC was more responsive after one night of sleep deprivation than after normal sleep. Increased subjective sleepiness in sleep-deprived subjects correlated significantly with activation of the PFC. The temporal lobe was activated after normal sleep but not after sleep deprivation; in contrast, the parietal lobes were not activated after normal sleep but were activated after sleep deprivation. Although sleep deprivation significantly impaired free recall compared with the rested state, better free recall in sleep-deprived subjects was associated with greater parietal lobe activation. These findings show that there are dynamic, compensatory changes in cerebral activation during verbal learning after sleep deprivation and implicate the PFC and parietal lobes in this compensation. PMID- 10688202 TI - The organizer factors Chordin and Noggin are required for mouse forebrain development. AB - In mice, there is evidence suggesting that the development of head and trunk structures is organized by distinctly separated cell populations. The head organizer is located in the anterior visceral endoderm (AVE) and the trunk organizer in the node and anterior primitive streak. In amphibians, Spemann's organizer, which is homologous to the node, partially overlaps with anterior endoderm cells expressing homologues of the AVE markers cerberus, Hex and Hesx1. For mice, this raises the question of whether the AVE and node are independent of each other, as suggested by their anatomical separation, or functionally interdependent as is the case in amphibians. Chordin and Noggin are secreted bone morphogenetic protein (BMP) antagonists expressed in the mouse node, but not in the AVE. Here we show that mice double-homozygous mutants that are for chordin and noggin display severe defects in the development of the prosencephalon. The results show that BMP antagonists in the node and its derivatives are required for head development. PMID- 10688203 TI - Maintenance of functional equivalence during paralogous Hox gene evolution. AB - Biological diversity is driven mainly by gene duplication followed by mutation and selection. This divergence in either regulatory or protein-coding sequences can result in quite different biological functions for even closely related genes. This concept is exemplified by the mammalian Hox gene complex, a group of 39 genes which are located on 4 linkage groups, dispersed on 4 chromosomes. The evolution of this complex began with amplification in cis of a primordial Hox gene to produce 13 members, followed by duplications in trans of much of the entire unit. As a consequence, Hox genes that occupy the same relative position along the 5' to 3' chromosomal coordinate (trans-paralogous genes) share more similarity in sequence and expression pattern than do adjacent Hox genes on the same chromosome. Studies in mice indicate that although individual family members may have unique biological roles, they also share overlapping functions with their paralogues. Here we show that the proteins encoded by the paralogous genes, Hoxa3 and Hoxd3, can carry out identical biological functions, and that the different roles attributed to these genes are the result of quantitative modulations in gene expression. PMID- 10688204 TI - The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. AB - Campylobacter jejuni, from the delta-epsilon group of proteobacteria, is a microaerophilic, Gram-negative, flagellate, spiral bacterium-properties it shares with the related gastric pathogen Helicobacter pylori. It is the leading cause of bacterial food-borne diarrhoeal disease throughout the world. In addition, infection with C. jejuni is the most frequent antecedent to a form of neuromuscular paralysis known as Guillain-Barre syndrome. Here we report the genome sequence of C. jejuni NCTC11168. C. jejuni has a circular chromosome of 1,641,481 base pairs (30.6% G+C) which is predicted to encode 1,654 proteins and 54 stable RNA species. The genome is unusual in that there are virtually no insertion sequences or phage-associated sequences and very few repeat sequences. One of the most striking findings in the genome was the presence of hypervariable sequences. These short homopolymeric runs of nucleotides were commonly found in genes encoding the biosynthesis or modification of surface structures, or in closely linked genes of unknown function. The apparently high rate of variation of these homopolymeric tracts may be important in the survival strategy of C. jejuni. PMID- 10688205 TI - Shiga-like toxins are neutralized by tailored multivalent carbohydrate ligands. AB - The diseases caused by Shiga and cholera toxins account for the loss of millions of lives each year. Both belong to the clinically significant subset of bacterial AB5 toxins consisting of an enzymatically active A subunit that gains entry to susceptible mammalian cells after oligosaccharide recognition by the B5 homopentamer. Therapies might target the obligatory oligosaccharide-toxin recognition event, but the low intrinsic affinity of carbohydrate-protein interactions hampers the development of low-molecular-weight inhibitors. The toxins circumvent low affinity by binding simultaneously to five or more cell surface carbohydrates. Here we demonstrate the use of the crystal structure of the B5 subunit of Escherichia coli O157:H7 Shiga-like toxin I (SLT-I) in complex with an analogue of its carbohydrate receptor to design an oligovalent, water soluble carbohydrate ligand (named STARFISH), with subnanomolar inhibitory activity. The in vitro inhibitory activity is 1-10-million-fold higher than that of univalent ligands and is by far the highest molar activity of any inhibitor yet reported for Shiga-like toxins I and II. Crystallography of the STARFISH/Shiga-like toxin I complex explains this activity. Two trisaccharide receptors at the tips of each of five spacer arms simultaneously engage all five B subunits of two toxin molecules. PMID- 10688206 TI - How self-tolerance and the immunosuppressive drug FK506 prevent B-cell mitogenesis. AB - Therapy for transplant rejection, autoimmune disease and allergy must target mature lymphocytes that have escaped censoring during their development. FK506 and cyclosporin are immunosuppressants which block three antigen-receptor signalling pathways (NFAT, NFkappaB and JNK), through inhibition of calcineurin, and inhibit mature lymphocyte proliferation to antigen. Neither drug induces long lived tolerance in vivo, however, necessitating chronic use with adverse side effects. Physiological mechanisms of peripheral tolerance to self-antigens provide an opportunity to emulate these processes pharmacologically. Here we use gene-expression arrays to provide a molecular explanation for the loss of mitogenic response in peripheral B-cell anergy, one aspect of immunological tolerance. Self-antigen induces a set of genes that includes negative regulators of signalling and transcription but not genes that promote proliferation. FK506 interferes with calcium-dependent components of the tolerance response and blocks an unexpectedly small fraction of the activation response. Many genes that were not previously connected to self-tolerance are revealed, and our findings provide a molecular fingerprint for the development of improved immunosuppressants that prevent lymphocyte activation without blocking peripheral tolerance. PMID- 10688207 TI - Localization of apical epithelial determinants by the basolateral PDZ protein Scribble. AB - The generation of membrane domains with distinct protein constituents is a hallmark of cell polarization. In epithelia, segregation of membrane proteins into apical and basolateral compartments is critical for cell morphology, tissue physiology and cell signalling. Drosophila proteins that confer apical membrane identity have been found, but the mechanisms that restrict these determinants to the apical cell surface are unknown. Here we show that a laterally localized protein is required for the apical confinement of polarity determinants. Mutations in Drosophila scribble (scrib), which encodes a multi-PDZ (PSD-95, Discs-large and ZO-1) and leucine-rich-repeat protein, cause aberrant cell shapes and loss of the monolayer organization of embryonic epithelia. Scrib is localized to the epithelial septate junction, the analogue of the vertebrate tight junction, at the boundary of the apical and basolateral cell surfaces. Loss of scrib function results in the misdistribution of apical proteins and adherens junctions to the basolateral cell surface, but basolateral protein localization remains intact. These phenotypes can be accounted for by mislocalization of the apical determinant Crumbs. Our results show that the lateral domain of epithelia, particularly the septate junction, functions in restricting apical membrane identity and correctly placing adherens junctions. PMID- 10688208 TI - A tripeptide 'anticodon' deciphers stop codons in messenger RNA. AB - The two translational release factors of prokaryotes, RF1 and RF2, catalyse the termination of polypeptide synthesis at UAG/UAA and UGA/UAA stop codons, respectively. However, how these polypeptide release factors read both non identical and identical stop codons is puzzling. Here we describe the basis of this recognition. Swaps of each of the conserved domains between RF1 and RF2 in an RF1-RF2 hybrid led to the identification of a domain that could switch recognition specificity. A genetic selection among clones encoding random variants of this domain showed that the tripeptides Pro-Ala-Thr and Ser-Pro-Phe determine release-factor specificity in vivo in RF1 and RF2, respectively. An in vitro release study of tripeptide variants indicated that the first and third amino acids independently discriminate the second and third purine bases, respectively. Analysis with stop codons containing base analogues indicated that the C2 amino group of purine may be the primary target of discrimination of G from A. These findings show that the discriminator tripeptide of bacterial release factors is functionally equivalent to that of the anticodon of transfer RNA, irrespective of the difference between protein and RNA. PMID- 10688209 TI - Pancreatic trauma--injuries to the pancreas and pancreatic duct. PMID- 10688210 TI - Organisation of a trauma registry in a regional Greek university hospital: the first two years experience. AB - OBJECTIVE: To design and implement a hospital trauma registry so as to be able to monitor the care of injured patients. SETTING: Teaching hospital, Greece. SUBJECTS: All patients admitted with trauma from January 1997. MAIN OUTCOME MEASURES: Design of a suitable form, establishment of inclusion and exclusion criteria, injury severity scoring, finding money and personnel, and getting suitable computer hardware and software for reliable collection and analysis of data. RESULTS: We experienced great difficulty in getting funding, so were unable to employ dedicated staff to collect the data, though we have had a part-time secretary to coordinate the registry whose salary has been paid by a pharmaceutical company. We have to rely on junior doctors to collect the data, which works well when they are enthusiastic (though not all are). We decided to use the data collection form used by the UK Trauma Network. We are trying to collect sufficient data to code severity by more than one system, but at present this is causing problems because busy nurses and doctors do not like filling in forms. Software has also been a problem as most of it is in English and translation is a considerable workload. The calculations are still being done manually while we work with two computer programmers to develop our own. We have submitted a research protocol to the Ministry of Health which has been accepted and this will guarantee our expenses for the next two years. CONCLUSIONS: Implementing a philosophy of continuous quality improvement is never easy, and we expect funding to be a permanent source of anxiety. Our progress so far has been good, but not as good as we hoped; however, we are optimistic that as people see the value of continuous monitoring of the system they will become more enthusiastic and supportive. PMID- 10688211 TI - Cardiac injuries: a ten-year experience. AB - OBJECTIVE: To present our experience of cardiac injuries treated at one Swedish emergency department in the 10 years 1988-97. DESIGN: Retrospective study. SETTING: Teaching hospital. SUBJECTS: 11 patients (9 men and 2 women, mean age 33 years, range 19-54); in 7 they were penetrating injuries and in 4 blunt. MAIN OUTCOME MEASURES: Morbidity and mortality. RESULTS: The mechanisms of injury were stab wound (n = 7), and car crash, fall, boat crash, and abuse (n = 1 each); drug or alcohol misuse played a part in all those with penetrating injuries. The penetrating wounds involved the left ventricle (n = 3), the right ventricle (n = 2), and the pericardium (n = 2). All 5 patients with ventricular wounds presented with cardiac tamponade, in 1 of whom it was fatal (he bled to death during emergency thoracotomy). The main complications were anoxic brain damage and postpericardiotomy syndrome (1 each). There was no case of myocardial concussion. CONCLUSION: Our data reflect the Swedish experience of heart trauma: there are few cases, alcohol and drug misuse is the principal risk factor, and there were no gunshot wounds. PMID- 10688212 TI - Outcome of lung trauma. AB - OBJECTIVE: To find out whether we could manage critical pulmonary haemorrhages in penetrating injuries, and to report our experience with blunt trauma of the lung. DESIGN: Retrospective study. SETTING: Teaching hospital, Sweden. SUBJECTS: 81 patients who presented with pulmonary injuries during the period January 1988 December 1997; 6 were penetrating and 75 blunt. RESULTS: There was only one patient with an isolated lung contusion. The remaining was divided into 2 groups: those with pulmonary contusion and thoracic lesions (n = 32), and those with pulmonary contusion and extrathoracic lesions (n = 42). Four patients in the penetrating group were shocked and required urgent operations; emergency room thoracotomy (n = 1), urgent thoracotomy (n = 2), and urgent thoracoabdominal exploration (n = 1) were done successfully. We correlated grade of lung injury [American Association for the Surgery of Trauma-Abbreviated Injury Scale (AIS)] with mortality. All patients with penetrating injuries survived without serious consequences. There were a mean (SD), of 6 (2) injuries/patient in those with extrathoracic injuries compared with 3 (1) injuries/patient in the group with thoracic lesions (p < 0.001). The corresponding hospital mortality was 6/42 (19%) mainly as a result of the central nervous system lesions (4/6) compared with 0/32. The mean (SD) Injury Severity Score (ISS) was 9.3 (4.8) in patients with thoracic lesions compared with 24.1 (14.7) in patients with extrathoracic lesions (p < 0.0001), and 14.9 (9.5) in all survivors compared with 49.9 (13.6) among those who died (p < 0.0001). CONCLUSIONS: An excellent outcome can be achieved managing penetrating injuries of the lung by an aggressive approach and urgent surgical intervention even when emergency room thoracotomy is essential. Pulmonary contusion is considered to be a relatively benign lesion that does not add to the morbidity or mortality in patients with blunt chest trauma. These data may help to decrease the obsession with pulmonary contusion in patients with chest trauma, with or without extrathoracic lesions, and avoid many unnecessary computed tomograms of the chest. PMID- 10688213 TI - Prognostic factors in patients with differentiated thyroid carcinoma. AB - OBJECTIVE: To study the prognostic factors in patients with differentiated thyroid carcinoma. DESIGN: Retrospective analysis. SETTING: University hospital, Germany. PATIENTS: 139 consecutive patients who underwent surgery for follicular (n = 42) and papillary thyroid carcinoma (n = 97). MAIN OUTCOME MEASURES: Survival rate, type of operation (systematic lymphadenectomy or no lymphadenectomy). RESULTS: Median observation time was 72 months (range 1-203). The 5 and 10 year survival rates in patients with papillary carcinoma were 92% and 89% respectively, and in those with follicular carcinoma 88% and 80%, respectively. Prognostic factors for papillary carcinoma were distant metastases, age, and extrathyroidal growth, and for follicular carcinoma they were distant metastases, extrathyroidal extension, and multifocal growth. The Union International contre le Cancer and European Organisation for Research and Treatment of Cancer scores and the age, grade, extent and size score were all highly significant. The extent of lymphadenectomy, primary or secondary thyroidectomy, and partial or total thyroidectomy did not influence survival. CONCLUSION: Staging and score systems may be helpful in calculating prognosis in differentiated thyroid carcinoma, but the benefit of systematic lymphadenectomy remains controversial. PMID- 10688214 TI - Clinical and histological differences in anaplastic thyroid carcinoma. AB - OBJECTIVE: To report our experience in patients with anaplastic thyroid carcinoma and try to establish differences between cases in which the histological study showed that there was an associated thyroid carcinoma and those that were strictly anaplastic or pure. DESIGN: Retrospective study. SETTING: University hospital, Spain. SUBJECTS: 14 patients with anaplastic thyroid cancer treated over a period of 26 years; 7 presented with associated thyroid tumours and 7 were pure. MEAN OUTCOME MEASURES: Clinical data (age, sex, symptoms), treatment, histological study (associated thyroid disease, spread, involved lymph nodes) and follow-up. RESULTS: 13 of the 14 tumours had spread locally. 5 patients were treated by total thyroidectomy, 3 subtotal thyroidectomy, 5 excision of the tumour, and 1 patient had a biopsy alone. There were associated thyroid tumours in 7 cases: 2 follicular, 2 tall cell papillary, 1 solid papillary, 1 medullary and 1 Hurthle cell tumour. 12 patients died. Another 2 are still alive having survived 61 and 70 months respectively, both with associated anaplastic cancers (follicular and solid). The mean survival was 14 months (24 for associated anaplastic carcinoma and 4 for pure anaplastic carcinoma). CONCLUSION: There is a subgroup of anaplastic cancers in which a better differentiated thyroid carcinoma coexists with the anaplastic carcinoma. The prognosis in this subgroup is better than that for primary pure anaplastic carcinoma. PMID- 10688215 TI - Upper gastrointestinal contrast study in the management of small bowel obstruction--a prospective randomised study. AB - OBJECTIVE: To find out whether contrast radiography helps to resolve small bowel obstruction. DESIGN: Prospective randomised trial. SETTING: University hospital, Norway. SUBJECTS: 98 consecutive patients with symptoms of small bowel obstruction and a plain abdominal radiograph that confirmed the diagnosis. INTERVENTIONS: The patients were randomly allocated to receive a mixture of barium and sodium diatrizoate (Gastrografin) (n = 48) or not (n = 50). Both groups were followed up clinically and by repeated abdominal films. MAIN OUTCOME MEASURES: Non-operative resolution of small bowel obstruction; number of patients with strangulated bowel; bowel resections; mortality; complications; hospital stay; and time from admission to operation. RESULTS: No significant differences were observed between the groups in the incidence of non-operative resolution (31/48 in contrast group, 35/50 in control group, OR: 0.89), strangulation obstruction (1/48 in contrast group, 4/50 in control group, OR: 0.24), bowel resection (3/48 in contrast group, 4/50 in control group, OR: 0.76), complications (8/48 in contrast group, 5/50 in control group, OR: 1.80), mortality (3/48 in contrast group, 1/50 in control group, OR: 3.26), and hospital stay (0-7 days: 34/48 in contrast group, 38/50 in control group, p = 0.95). The contrast group had a shorter interval between admission and operation than the control group (0-24 hours: 12/48 in contrast group, 3/50 in control group, p = 0.005). CONCLUSION: The contrast examination did not contribute to the resolution of small bowel obstruction. PMID- 10688216 TI - Open management of the abdomen and planned reoperations in severe bacterial peritonitis. AB - OBJECTIVE: To assess the results of open management of the abdomen and planned re operations in severe bacterial peritonitis after perforation or anastomotic disruption of the digestive tract. DESIGN: Retrospective study. SETTING: University Hospital, The Netherlands. SUBJECTS: 67 consecutive patients. INTERVENTIONS: Open management of the abdomen and planned reoperations. MAIN OUTCOME MEASURES: Hospital morbidity and mortality, long-term follow-up. RESULTS: 38 patients developed multiple organ failure (MOF), but 29 needed only ventilatory and inotropic support. The mean number of re-operations was nine. 16 patients developed severe bleeding and 16 fistulas. In-hospital mortality was 42% (n = 28). Long-term morbidity, particularly the number of abdominal wall defects (n = 10), was considerable. CONCLUSION: Despite open management of the abdomen and planned re-operations, mortality of severe bacterial peritonitis still continues to be too high, and both short and long-term morbidity are appreciable. The value of open management of the abdomen and planned re-operations rests only on the clinical observation that other conventional surgical treatments of severe bacterial peritonitis often fail. PMID- 10688217 TI - Transoral application of EEA stapler after subtotal oesophagectomy. AB - OBJECTIVE: To describe a new transoral technique of cervical oesophagogastric and oesophagojejunal anastomoses using the EEA stapler. DESIGN: Prospective clinical study. SETTING: University Hospital, Bratislava, Slovakia. SUBJECTS: Two patients with squamous cell carcinoma of the middle and distal third of the oesophagus. INTERVENTIONS: Transhiatal subtotal oesophagectomy without thoracotomy, and cervical oesophageal anastomosis by transoral EEA stapling. MAIN OUTCOME MEASURES: Morbidity and mortality. RESULTS: Transoral stapling was successful in both patients with no anastomotic leaks. The patients were discharged on the 14th and 21st postoperative days, respectively. CONCLUSIONS: Transoral stapling of the cervical anastomosis gave good results in two patients. More development and evaluation are needed. PMID- 10688218 TI - Inflammatory fibroid polyp and Helicobacter pylori. Aetiology or coincidence? AB - OBJECTIVE: To evaluate our incidence of inflammatory fibroid polyps, compare our experience with that of others, and to analyze the possible pathophysiological and aetiological factors. DESIGN: Retrospective review. SETTING: Teaching hospital. MATERIAL: All histopathological slides of the gastrointestinal tract. MAIN OUTCOME MEASURES: Incidence and treatment in our Medical Center and elsewhere. RESULTS: We could find only one case of inflammatory fibroid polyp, an estimated incidence of 1/4000. Between 1987-1996 only 331 were reported elsewhere, most of which (293, 88.5%), were located in the stomach. CONCLUSION: Primary mucosal damage can expose the stroma to several irritants (chemical, mechanical and biological), that may subsequently cause inflammatory fibroid polyps in certain people. PMID- 10688219 TI - Long term results of lateral pancreaticojejunostomy for chronic alcoholic pancreatitis. AB - OBJECTIVE: To assess our long term results of lateral pancreaticojejunostomy in patients with alcoholic pancreatitis. DESIGN: Retrospective study. SETTING: University hospital, France. SUBJECTS: 57 patients (48 men, 9 women, mean (SD) age 46 (7) years who required surgical treatment of chronic alcoholic pancreatitis between January 1977 and October 1995. INTERVENTIONS: Lateral pancreaticojejunostomy with or without another procedure. Outcome classified as excellent, good, fair, or poor. MAIN OUTCOME MEASURES: Postoperative morbidity and mortality; relief of pain; reduction in use of analgesics and exocrine supplements; effect on exocrine and endocrine insufficiency; and return to paid work. RESULTS: There were no postoperative deaths and no pancreatic fistulae, but there were 17 other postoperative complications (30%). Median follow up was 65 months (range 8-206), during which 12 patients died (21%). Result was judged excellent in 16 (28%), good in 27 (47%), fair in 5 (9%), and poor in 9 (16%). Pain control was significantly improved, analgesic usage decreased, less pancreatic enzyme supplementation was required, and 25 patients returned to paid work (p = 0.0001 in each case). Exocrine and endocrine function remained stable. The results were better if the patient gave up misusing alcohol (p = 0.03) and if the operation was done within 4 years of the development of pancreatitis (p = 0.03). CONCLUSIONS: Lateral pancreaticojejunostomy is a safe procedure that can improve functional outcome in patients with chronic alcoholic pancreatitis, and does not worsen pancreatic function. PMID- 10688220 TI - Operative monitoring of hand and axillary temperature during endoscopic superior thoracic sympathectomy for the treatment of palmar hyperhidrosis. AB - OBJECTIVE: To find out how much the temperature in the palm rises after upper thoracic sympathectomy for palmar hyperhidrosis, and correlate the temperature with the outcome. DESIGN: Retrospective study. SETTING: University hospital, Spain. SUBJECTS: 73 patients (34 women and 39 men, age range 16-42 years, mean 26) who were operated for palmar hyperhidrosis between 1 January 1995 and 31 December 1997. INTERVENTIONS: Bilateral thoracic endoscopic sympathectomy during which the temperature was monitored on the skin of both axillae and thenar eminences, and in the oesophagus. MAIN OUTCOME MEASURES: Morbidity, alleviation of hyperhidrosis, recurrence rate, and differences in temperature postoperatively. RESULTS: There was minor bleeding during operation in 25 cases (34%), but in only 4 was it sufficient to require insertion of a drain; 2 patients developed transient Homer's syndrome; but the most common complication was compensatory hyperhidrosis (n = 52, 71%). In only 5 was this other than mild and required treatment with aluminium chloride in ethanol 25%. Palmar hyperhidrosis was alleviated in all cases, axillary sweating was considerably improved, and there was improvement in the feet in 56 (77%). There were 5 recurrences, all on the right side, during a mean follow up of 9 months (range 2 36), but in no case was the sweating severe. In almost all cases the temperature of the palm was less than that of the axilla before operation by a mean (SD) of 0.9 (0.3) degrees C. The rise in temperature varied from 1.7 (0.4) degrees C to 2.6 (0.4) degrees C. In the 5 patients who developed recurrences the increase was less (0.5 (0.4) degrees C). CONCLUSION: Thoracic endoscopic sympathectomy is safe, simple, and effective in treating palmar hyperhidrosis that has not responded to conservative treatment. Intradermal monitoring is an accurate and cost-effective way of monitoring temperature during operation. Although it is essential to achieve a rise in temperature of 1 degrees C, our most important finding was that the final temperature in both hands and axillae should be above 35 degrees C and as near as possible to 36 degrees C. PMID- 10688221 TI - Intestinal perfusion during pneumoperitoneum with carbon dioxide, nitrogen, and nitric oxide during laparoscopic surgery. AB - OBJECTIVE: To find out what effect insufflation pressure and type of gas have on intestinal perfusion during pneumoperitoneum. DESIGN: Randomized, controlled, prospective, experimental study. SETTING: University affiliated animal experimental laboratory, Sweden. ANIMALS: Fasted, anaesthetised, domestic pigs of both sexes operated on laparoscopically (n = 7, weight 26-31 kg). INTERVENTIONS: Insufflation of carbon dioxide (CO2), nitric oxide (NO), or nitrogen (N2) at intra-abdominal pressures of 0, 5, 10, 15 and 20 mm Hg. MAIN OUTCOME MEASURES: Cardiac output, portal blood flow, and jejunal mucosal perfusion. RESULTS: Cardiac output decreased during N2 and NO (15, 20 mm Hg) but not during CO2 insufflation because of an accompanying tachycardia. Portal flow decreased during insufflation with N2 and NO (15, 20 mm Hg) and CO2 (20 mm Hg). Jejunal perfusion was reduced during N2 and NO insufflation (5-20 mm Hg) but remained unchanged during CO2 insufflation (5-20 mm Hg). CONCLUSIONS: Insufflation with CO2 maintained jejunal mucosal perfusion, probably as a result of hypercarbia as N2 at equal pressures reduced mesenteric flow. The vasodilator NO provided no haemodynamic benefit. PMID- 10688222 TI - Improvement of portal flow and hepatic microcirculatory tissue flow with N acetylcysteine in dogs with obstructive jaundice produced by bile duct ligation. AB - OBJECTIVE: To find out if N-acetylcysteine (NAC) would improve hepatic circulation in dogs with obstructive jaundice. DESIGN: Open laboratory study. SETTING: University hospitals, Japan and France. MATERIALS: 14 male beagle dogs and 10 male Wistar rats. INTERVENTIONS: Obstructive jaundice was produced by ligation of the common bile duct (CBD) for 7 days in both dogs and rats. Either 5% dextrose (control group, n = 7) or NAC (NAC group, n = 7) was given to dogs. Sinusoidal endothelial cells were obtained from rats after ligation by elutriation, and varying amounts of NAC were given. MAIN OUTCOME MEASURES: The volumes of portal blood flow and hepatic microcirculatory tissue flow were reduced after ligation of the CBD, but those increased after NAC had been given to dogs with obstructive jaundice. NAC increased the concentrations of plasma cyclic 3',5'-guanosine monophosphate (cGMP). It also increased concentrations of serum and hepatic-reduced glutathione, and hepatic adenosine triphosphate (ATP) in cholestatic dogs, and secretion of cGMP from sinusoidal endothelial cells from rats with obstructive jaundice. CONCLUSION: These results suggest that NAC given intravenously effectively improves hepatic circulation and hepatic function in dogs with obstructive jaundice. PMID- 10688223 TI - Direct insertion of a hepatic arterial catheter after hepatectomy. PMID- 10688224 TI - Pancreatic foregut cyst. PMID- 10688225 TI - 'Body packer' syndrome: characteristics and treatment--case report and review. PMID- 10688226 TI - Solitary sternal breast cancer metastases treated by sternectomy and muscle flap reconstruction. PMID- 10688227 TI - Some plane truths about pictures: notes on Wagemans, Lamote, and van Gool (1997). AB - Wagemans, Lamote, and van Gool (1997) have attempted to show that observers can determine the geometric equivalence of shapes seen in perspective, or in projective transformation. Artifacts of measurement in their procedures forestall such a conclusion. Their experiment fails to control projective properties adequately, and also confounds the transformations of shear and compression. Their evidence that observers can discern the equivalence of shapes under perspective can be reconstrued as evidence for sensitivity to different, unrelated properties: to the plane compression that follows from the depiction of flat shapes in perspective, and to gross differences in shape not specific to projective geometry. An improved set of procedures is proposed for the measurement of stimuli in the study of visual shape constancy. PMID- 10688228 TI - Pre-attentive segmentation in the primary visual cortex. AB - The activities of neurons in primary visual cortex have been shown to be significantly influenced by stimuli outside their classical receptive fields. We propose that these contextual influences serve pre-attentive visual segmentation by causing relatively higher neural responses to important or conspicuous image locations, making them more salient for perceptual pop-out. These locations include boundaries between regions, smooth contours, and pop-out targets against backgrounds. The mark of these locations is the breakdown of spatial homogeneity in the input. for instance, at the border between two texture regions of equal mean luminance. This breakdown causes changes in contextual influences, often resulting in higher responses at the border than at surrounding locations. This proposal is implemented in a biologically based model of VI in which contextual influences are mediated by intra-cortical horizontal connections. The behavior of the model is demonstrated using examples of texture segmentation, figure-ground segregation, target-distractor asymmetry, and contour enhancement, and is compared with psychophysical and physiological data. The model predicts (1) how neural responses should be tuned to the orientation of nearby texture borders, (2) a set of qualitative constraints on the structure of the intracortical connections, and (3) stimulus-dependent biases in estimating the locations of the region borders by pre-attentive vision. PMID- 10688229 TI - The roles of polarity and symmetry in the perceptual grouping of contour fragments. AB - We describe two experiments that investigate the roles of polarity and symmetry in the perceptual grouping of contour fragments. Observers viewed, for one second on each presentation, arrays of oriented, spatial-frequency band-pass, elements, in which a subset of the elements was aligned along a twisting curve. In each of five conditions we measured observers' ability to detect aligned combinations of even- and odd-symmetric elements, of the same and different polarities, against a background of 'noise' elements. As with previous experiments we found that the 'path' could be reliably detected, even when the elements of the path were oriented at angles of up to +/- 60 deg relative to each other. Detection of the path was still possible when the polarity of path elements alternated. However, the probability of detection of the path was raised significantly when the path elements were all of the same polarity. Perceptual grouping of even-symmetric elements was no different to perceptual grouping of odd-symmetric elements. The results provide evidence, that in achieving integration of contour fragments, the visual system uses a process that is to some degree phase selective. We use the results to describe how the visual system may resolve natural contours when they occur against backgrounds that vary over a wide range of intensities. The data presented here have been published in conference-abstract form (Hayes et al., 1993; Field et al., 1997). PMID- 10688230 TI - Perceived motion in complementary afterimages: verification of a neural network theory. AB - Steady fixation of a regular pattern like a bar grating or concentric circles leads to a complementary afterimage at pattern offset. The afterimage has the appearance of shimmering lines that are locally orthogonal to the orientations of the inducing image. Additionally, the afterimage includes motion running parallel to the orientation of the afterimage lines. We argue that this afterimage motion supports the existence of a cue to motion that is based on the spatial organization of oriented responses. This cue was previously proposed after analysis of a neural network model of visual perception. We test predictions of the model on various types of complementary afterimage inducing stimuli. When a contrast or size gradient is included in the inducing image, the afterimage motion moves toward the higher part of the gradient, in agreement with the model. Implications of this cue for computational and neurophysiological theories of motion perception are discussed. PMID- 10688231 TI - Conjunctions of colour, luminance and orientation: the role of colour and luminance contrast on saliency and proximity grouping in texture segregation. AB - To examine whether perceptual grouping on the basis of orientation can be performed simultaneously with or only subsequently to grouping according to colour or luminance, we tested whether subjects are able to segregate arrays of texture elements that differ from surrounding elements by conjunctions of either (i) colour and orientation, or (ii) luminance contrast and orientation, or (iii) luminance contrast polarity and orientation. Subjects were able to use conjunctions between luminance and orientation for segregation but not conjunctions between colour or contrast polarity and orientation. Our results suggest that (i) in agreement with earlier findings, there seem to exist no specific conjunction detectors for colour and orientation or contrast polarity and orientation, and (ii) when orientation defined textures are to be distinguished by virtue of differences in luminance, colour, or contrast polarity, luminance provides a much stronger cue than colour or contrast polarity for saliency-based orientation grouping. PMID- 10688232 TI - Tilt aftereffects generated by bilaterally symmetrical patterns. AB - Tilt aftereffects were generated by bilaterally symmetrical dot patterns. Both expansion and contraction effects, similar in size and magnitude to effects usually reported with luminance contours, were observed after adaptation to symmetrical patterns tilted 15 deg or 75 deg respectively from a vertically oriented test. Large effects were found when both adapting and test stimuli were symmetrical patterns while smaller effects were found when the adapting stimulus was symmetrical and test stimulus was a grating. A third experiment, which manipulated the number of dots near the axis line, confirmed the above findings; expansion and contraction effects were observed again. The results of these experiments suggest that the neural mechanism underlying the perception of luminance contours may be linked to the mechanism for the detection of symmetry. PMID- 10688233 TI - Risk-stratification strategy eludes error researchers. PMID- 10688234 TI - Studies published on clopidogrel-aspirin for coronary stent placement. PMID- 10688235 TI - Higher copayment does not hurt compliance, study suggests. PMID- 10688236 TI - HMOs pressed by rising drug costs, industry report shows. PMID- 10688237 TI - Partnering with nurses to manage heparin therapy with a weight-based protocol. PMID- 10688238 TI - SAMe: S-Adenosylmethionine. PMID- 10688239 TI - New drug overview. Pioglitazone hydrochloride. PMID- 10688240 TI - Making a difference in hypertension. PMID- 10688241 TI - Projecting future drug expenditures--2000. AB - Drug cost projections for 2000 and factors that are likely to influence drug costs are discussed. The total increase in drug costs in 2000, assuming there are no major unforeseen events (e.g., Medicare coverage of outpatient prescriptions or important new drugs) can be anticipated to be 12-15% above that in 1999. Prescription drug expenditures are projected to increase an average of 11.2% annually between 1999 and 2001. Although there were fewer new drug approvals in the first half of 1999 than in 1998, we can expect that many new, important, and probably expensive agents will be coming to market in 2000. Analysts calculate that pharmaceutical companies invested approximately $24 billion in research and development in 1999, and there are predictions that mergers within the pharmaceutical industry will reduce the number of major pharmaceutical firms to 13. The introduction of pharmacogenomics is of great interest because of its potential to significantly affect drug therapy and the discovery of new agents. Sales of generic drugs are anticipated to increase from $12 billion in 1999 to $14 billion in 2000. Competition between the brand-name and generic drug industries is intensifying, but it remains unclear whether generic drug pricing will offset brand-name price increases. Internet pharmacy has become a reality and is under federal and state scrutiny. The pharmaceutical and drug distribution industries are changing dramatically, and there is concern that many new prescription drugs will not be readily available to patients because of their high cost. Should this occur, action by the federal government is a possibility. PMID- 10688242 TI - Compliance with and dosing of angiotensin-converting-enzyme inhibitors before and after hospitalization. AB - Compliance with and dosing of angiotensin-converting-enzyme (ACE) inhibitors as they occur before and after hospitalization for heart failure were studied, and factors predictive of compliance with and dosing of ACE inhibitors after hospitalization were identified. Two hundred thirty-six patients hospitalized with heart failure between October 1, 1995, and April 30, 1996, were identified. Compliance with and use and dosing of ACE inhibitors were examined over the 180 day period before admission and the 180-day period after discharge using an integrated pharmacy and medical claims database. Use of an ACE inhibitor was defined as at least one claim for an ACE inhibitor over the period examined, and dosing was assessed by calculating the mean percentage of an adequate daily dose dispensed. Before hospitalization 109 patients (46.2%) used ACE inhibitors, and after hospitalization 148 (62.7%) used them--a significant increase. ACE inhibitor use before hospitalization was a predictor of postdischarge use. Younger patients were more likely to take ACE inhibitors after hospitalization than older ones, and men had better compliance after hospitalization than women. Additional analyses revealed that, among hospitalized patients, compliance was lower in individuals who also took an antidepressant. Dosing increased from 72% to 85% of an adequate daily dose after hospitalization among patients who took ACE inhibitors during both prehospitalization and posthospitalization periods. However, almost one third of hospitalized patients stopped taking their ACE inhibitor within six months of hospital discharge. The study found few significant predictors of patient compliance after hospitalization. Dosing of ACE inhibitors before and after hospitalization needs to be improved. PMID- 10688243 TI - Applied pharmacoeconomics: modeling data from internal and external sources. AB - The use and application of techniques for modeling data obtained from various data sources are discussed. Modeling with internal and external data has become a popular way for health care organizations to apply pharmacoeconomics to pharmacy practice. Modeling studies use existing clinical and epidemiologic data to project the effect of a clinical, policy, or medication decision on a patient, population, or organization. Although several modeling techniques have been used in health care, the most common approaches are to modify and adapt existing models or to develop a unique model to answer questions of interest in a specific practice setting. Typically, an economic model developed by adapting an existing one will use either the clinical decision-analysis or Markov modeling technique. Regardless of the technique used, external data must be carefully evaluated to ensure that the data are appropriate for use in making decisions at a specific organization. For example, cost data must be analyzed to ensure that the calculations are reproducible. Also, it must be acknowledged that this strategy may not always be appropriate. The use of modeling techniques can assist decision makers in making more informed clinical, policy, and medication decisions in real world settings. Caution is required when adapting and interpreting existing models to ensure appropriate application in a specific organization. PMID- 10688244 TI - Clinical practice guidelines and the standard of care. PMID- 10688245 TI - ASHP therapeutic position statement of optimizing treatment of hypertension. AB - The JNC-VI report provides solid recommendations for optimizing the treatment of hypertension. Suggestions for when to initiate lifestyle modifications and drug therapy on the basis of risk stratification are major improvements in the current report. Although diuretics and beta-blockers are still considered the preferred agents for all patients with uncomplicated hypertension, individualization of therapy when there are complications or coexisting conditions is strongly encouraged. If blood pressure control rates are to improve in the United States, aggressive approaches to the prevention, identification, management, and tracking of patients and populations with hypertension will be required. Pharmacists will be critical in these efforts to improve blood pressure control and will often need to collaborate with physicians in providing drug therapy management. All pharmacists who help manage patients with hypertension should be familiar with the entire JNC-VI report. PMID- 10688246 TI - Joint commission hospice survey for pharmaceutical services. PMID- 10688247 TI - Predictable positive bias with portable capillary blood monitors. PMID- 10688248 TI - Protection of labile pharmaceuticals. PMID- 10688249 TI - Evidence-based medicine: its application to laboratory medicine. AB - The current health care environment of cost-cutting highlights the need to reinforce the contribution of laboratory medicine to improvement in health care. This must be a patient-focused activity using continuous quality improvement, a familiar concept in laboratory practice. Involvement in the creation of clinical practice guidelines, care maps, and outcome measures will place laboratory medicine in the circle of continuous quality improvement. The laboratory must provide strong evidence that tests contribute to better overall resource utilization. Laboratory Information Systems can be used to better integrate laboratory data with clinical, diagnostic, pharmaceutic, statistical, and financial information. Improving laboratory utilization requires clear demonstrations of appropriate versus inappropriate laboratory use, and instructions on implementing appropriate use. The education of laboratory professionals should include search strategies, understanding the diagnostic accuracy of medical tests, and the application of systematic reviews and meta analysis. With the rapid increase in the data base supporting evidence-based laboratory medicine, there is a significant challenge in translating the existing knowledge into practice. There is also a need for a cooperative strategy between the diagnostics industry and the laboratory medicine profession to provide evidence of the added value of laboratory testing. There is a significant role in developing the academic basis of the unique aspects of evidence-based laboratory medicine. PMID- 10688250 TI - Identification of uridine diphosphate glucuronosyltransferases involved in the metabolism and clearance of mycophenolic acid. AB - Mycophenolic acid, the active metabolite of the immunosuppressant and antiproliferative agent, mycophenolate mofetil, is primarily metabolized by glucuronidation to the inactive 7-O-glucuronide. Although the uridine diphosphate (UDP) 7-O-glucuronide is the principal excretion product of this drug, carboxyl linked glucuronides have also been detected in vitro and in vivo. To identify human UDP glucuronosyltransferases that are active in the glucuronidation of mycophenolic acid, cDNAs encoding individual UDP glucuronosyltransferase forms have been expressed in cell culture, and the capacity of the expressed enzymes to use mycophenolic acid as a substrate has been assessed. Two UDP glucuronosyltransferase forms, UGT1A8 and UGT1A10, were active in the glucuronidation of mycophenolic acid. Both enzymes are predominantly expressed in the gastrointestinal tract and hence, may play a role in the metabolism of mycophenolic acid in the gastrointestinal tract and in the acquisition of resistance to the mito-inhibitory effects of this drug in cultured human colorectal carcinoma cell lines. The identities of the UDP glucuronosyltransferase forms that are mainly responsible for the glucuronidation of mycophenolic acid in the liver and kidney remain unknown; however, UGT1A9 may be important in this respect as the cDNA-expressed enzyme has some capacity to glucuronidate mycophenolic acid. Other UGT1A forms in the liver and kidney (UGT1A1, UGT1A3, UGT1A4, and UGT1A6) were inactive toward mycophenolic acid. PMID- 10688251 TI - Pharmacokinetics and concentration-control investigations of mycophenolic acid in adults after transplantation. AB - Data have emerged that provide the scientific basis for therapeutic drug monitoring of mycophenolic acid (MPA) in transplant patients receiving mycophenolate mofetil (MMF), the parent drug, in combination with other immunosuppressive agents. There is a significant relationship between the dose interval MPA AUC and risk for acute rejection based on retrospective investigations in renal and heart transplant patients and on prospective investigations in renal transplant patients. The MPA dose-interval AUC varies naturally by more than 10-fold in renal and heart transplant patients. Other significant sources of pharmacokinetic variability for MPA include the effects of concomitant medications, and the effects of disease states such as renal dysfunction and liver disease on the steady state MPA AUC. Individualized MMF dose evaluation, guided by MPA plasma concentrations, is becoming the standard of practice at a growing number of transplant centers worldwide because of these factors and because of the need to closely evaluate the immunosuppression afforded by MPA when a change in the immunosuppression regimen in stable transplant patients is planned. Investigations of therapeutic drug monitoring strategies with an emphasis on identifying an optimal abbreviated sampling strategy for MPA AUC estimation are ongoing. Based on the concentration-outcome studies and experience at the authors' institutions and other centers, the authors propose a set of therapeutic drug monitoring guidelines for MPA in stable renal and heart transplant patients for the immediate (first 3 months posttransplant) and maintenance (>3 months) periods. When MPA binding to human serum albumin is altered, as occurs in patients with significant renal dysfunction, liver disease, or a substantial reduction in human serum albumin concentration, the possibility of increased MPA free fraction and free concentration will need to be taken into account in the interpretation of MPA total concentrations. PMID- 10688252 TI - Pharmacokinetic and metabolic investigations of mycophenolic acid in pediatric patients after renal transplantation: implications for therapeutic drug monitoring. German Study Group on Mycophenolate Mofetil Therapy in Pediatric Renal Transplant Recipients. AB - The need for mycophenolic acid (MPA) monitoring is still under discussion. Key issues for the PK/PD relationships of this drug are: the role of metabolites, the usefulness of AUC versus predose levels, and the need to monitor the free concentration of MPA (f-MPA). Recent advances have revealed that, in addition to 7-O-MPAG, three additional MPA metabolites are present in the plasma of transplant recipients. One of these metabolites (M-2), identified as an acyl glucuronide of MPA, was found to inhibit IMPDH-II in vitro. This active metabolite was also found to cross-react in the Emit assay for MPA. In an ongoing multicenter study, the authors are evaluating the relevance of monitoring total (t-MPA) and free mycophenolic acid (f-MPA) in pediatric renal transplant recipients. As in adults, a time-dependent increase of t-MPA-AUC(0-12h) within the first 3 months posttransplant (35 versus 64 mg x h/L, [corrected] 3 weeks versus 3 months respectively; daily dosage: 0.6 g/m2 bid) was seen. Receiver operating characteristics curve analyses were used to test the ability of predose levels or AUC(0-12h) to discriminate between cases with no complications and those with acute rejection, adverse events (severe infections, leukopenia), or gastrointestinal disorders observed during the early posttransplant course. In agreement with observations in adults, a significant (p = 0.001) association was observed between AUC(0-12h) and acute rejection. A t-MPA-AUC(0-12h) of approximately 30-60 mg x h/L [corrected], as determined by HPLC, seems to be a reasonable target for the early posttransplant period. It remains to be elucidated whether regular predose level monitoring may be of more practical value. A higher incidence of rejection was observed at predose MPA concentrations < or = 1 mg/L, as measured by HPLC. In contrast to t-MPA, f-MPA-AUC(0-12h) was significantly related to severe infections and leukopenia. The risk for severe adverse events was increased at f-MPA- AUC(0-12h) values > or =600 microg x h/L [corrected]. On the basis of these data and the observed variability in the pharmacokinetics of MPA, the development of monitoring strategies for this drug appears to be promising. PMID- 10688253 TI - Pharmacokinetics of mycophenolic acid in renal insufficiency. AB - Mycophenolate mofetil (MMF) is now widely used in solid organ transplantation. MMF is rapidly converted to its active form, mycophenolic acid (MPA), upon reaching the systemic circulation. MPA is metabolized to its glucuronide metabolite, mycophenolic acid glucuronide (MPAG), by glucoronyl transferases in the liver and possibly elsewhere. MPAG is then excreted by the kidney. MPA is extensively and avidly bound to serum albumin. Previous studies have demonstrated that it is only the free (non-protein-bound) fraction of MPA that is available to exert its action. In vivo and in vitro studies demonstrate that renal insufficiency decreases the protein binding of MPA and increases free MPA concentrations. This decrease in protein binding seems to be caused both by the uremic state itself and by competition with the retained metabolite MPAG. The disposition of MPA in patients with severe renal impairment may be significantly affected by this change in protein binding. PMID- 10688254 TI - Pharmacokinetics and metabolism of sirolimus. AB - Sirolimus (rapamycin, Rapamune) is a potent immunosuppressive drug that received marketing approval from the US Food and Drug Administration on September 15, 1999. Research into defining its pharmacokinetic (PK) behavior, interaction with other agents, and metabolism is ongoing. It has been established that oral doses of both liquid and solid formulation are rapidly, though incompletely and variably, absorbed. Metabolism by the intestinal and hepatic CYP3A family of enzymes likely contributes to variability in absorption and low bioavailability. Sirolimus has a long terminal half-life, the AUC correlates well with trough and peak concentrations, and it exhibits a moderate degree of dose proportionality. There is significant interpatient variability in PK parameters of sirolimus, though it exhibits predictable PK behavior when used with prednisone and cyclosporine neoral. There is a decreased rejection risk with higher doses and target level attainment. Several species of sirolimus metabolites have been characterized, and are measurable in whole blood and tissue specimens. Many more species of sirolimus metabolites are detectable, but they are not quantifiable at this time. The total concentration of metabolites appears to be less than that of the parent drug when examined through the PK profile. A reference method for the quantitation of metabolites remains elusive because of a lack of proper standardization. The clinical significance of sirolimus metabolites remains to be proven. PMID- 10688255 TI - Pharmacoeconomics of therapeutic drug monitoring in transplantation. AB - Immunosuppressive drugs have contributed significantly to the success of organ transplantation. Therapeutic drug monitoring is an integral part of transplant protocols. However, there is little information concerning its positive contribution to pharmacoeconomics. Before developing studies to demonstrate the potential benefits of TDM, consideration must be given to the type of TDM to be evaluated. It is argued that, given that the lymphocyte in the central compartment is the target for immunosuppressants, Area-Under-the-Curve monitoring may be a better reflection of control and toxicity than traditional trough monitoring. PMID- 10688256 TI - Cytokines and their receptors: an overview. AB - Cytokines participate in the induction and effector phases of all immune and inflammatory responses. They are therefore obvious candidates for exploitation as drugs or drug targets to promote, limit, or alter these responses in infection, allergy, autoimmunity, and other disease states. Although some cytokines and related molecules are already in clinical use, the full therapeutic potential of this class of hormones has yet to be realized, and this will depend in part on understanding their normal functions and regulation in health and disease. An overview is given of the cytokines and their receptors, their genetic and structural relationships, and the regulation of their activities by naturally occurring antagonists and other physiologic mechanisms. Some of the ways in which new therapeutic strategies are being developed from knowledge of the structure, function, and regulation of these various components of the cytokine network are outlined. PMID- 10688257 TI - Immunophilins: their properties and their potential role in therapeutic drug monitoring. AB - This article reviews recent developments in the immunophilin arena and the current state of knowledge about the biochemical properties of immunophilins. The role of immunophilin-binding assays is also explored with the conclusion that some of these immunosuppressive-drug-binding proteins may provide better assays for sirolimus and tacrolimus than current immunoassays. PMID- 10688258 TI - The FTY720 story. AB - The chemical 2-amino-2[2-(4-octylphenyl)ethyl]-1,3,propane diol is one of a class of small-molecule immunosuppressive agents. Better known as FTY720, this compound was chemically synthesized in an effort to minimize the toxic in vivo properties of a structurally related and highly potent immunosuppressive agent, myriocin. FTY720's mechanism of action, although not fully characterized, appears to be unique among immunosuppressants. Whereas the most well known biochemical characteristic of myriocin is its ability to inhibit serine palmitoyl transferase, the enzyme that initiates the biosynthetic pathway that leads to sphingosine, FTY720 is ineffective in this regard. In vivo, FTY720 induces a significant reduction in the number of circulating lymphocytes. It is thought to act by altering lymphocyte trafficking/homing patterns through modulation of cell surface adhesion receptors and ligands in a manner that has yet to be elucidated. Although much research has yet to be done to unravel the nature of the mechanism of action of FTY720, its efficacy has been sufficiently proven in numerous animal models, especially when administered in combination with cyclosporine. The agent is now progressing through human clinical trials, with the results of phase 1 clinical trials showing safety and tolerability in adult recipients of renal transplants. It is hoped that FTY720 will eventually prove to be an efficacious new weapon in the immunosuppressive armamentarium. PMID- 10688259 TI - Challenges of toxicology for the millennium. AB - In meeting the challenges of toxicology, clinical and forensic toxicologists should expand their services and engage in research and development to meet changing needs. Expanding roles could potentially derive from the threat of terrorism, genotyping for interpretation of potential toxic drug interactions, and criminalistic testings. At the threshold of the next millennium, terrorism via weapons of mass destruction (WMD) has migrated from the war zones to civilian settings. These WMD may be in the form of nuclear, biological, and chemical devices (NBC). Recently, the possible use of chemical/biological weapons in the Middle East conflicts, the use of sarin in a Tokyo subway station, and the unregulated availability of nuclear fuel in some countries all have heightened the potential for international and domestic NBC. In preparation for NBC, both government and civilians in major American cities have been trained for safe handling of patients and casualties. Forensic and clinical toxicologists should be knowledgeable about the clinical pharmacology, safe samples processing, and possible screening and/or analysis of samples exposed to or containing: vesicants; cyanide; and nerve, riot control, and pulmonary agents. These samples may be transported for further analysis and confirmed by designated central laboratories. In criminal/correctional settings, toxicologists should engage in quality assurance and consultation with attorneys, judges, and correctional professionals. With the emergence of pharmacogenetics, genotyping may enhance rational drug therapy for enhanced patient care, and may explain adverse or fatal drug reactions in postmortem analysis. PMID- 10688260 TI - Aquatic toxicology. AB - Australia is blessed with a great diversity of unique species in its fresh waters and in the marine environment around its coast. There is evidence that human and natural events are impacting on these species. Such impacts are associated with various agricultural, industrial, and domestic practices and with natural and anthropogenically driven climate change. Among the species most affected are those living in aquatic and marine environments. Some of these, such as cyanobacteria and dinoflagellates, have the potential for toxicity. Linked to this, there is the potential benefit of harnessing the pharmacologic potential of these toxins. PMID- 10688261 TI - Ion channel toxins and therapeutics: from cone snail venoms to ciguatera. AB - Ion channels are intimately linked to all neurotransmission and neurotransmitter release processes, but in disease states often contribute adversely to disease pathology. The diversity and distribution of ion channel types and subtypes being uncovered through the use of molecular biology and toxin probes present an exciting opportunity for the discovery of new, more selective drugs. Among ion channels targeted by cone shell venom peptides (conotoxins) are the voltage sensitive sodium, calcium, and potassium channels which open and then close (inactivate) in response to membrane depolarization, and thus regulate neurotransmission and the neurotransmitter release process. Conotoxins also target ligand-gated ion channels, including the NMDA-glutamate channel and the nicotinic acetylcholine receptor channel. The diversity of subtypes, especially those subtypes upregulated in disease states, makes ion channels a rapidly expanding therapeutic area. Conotoxins represent some of the most selective inhibitors of ion channel subtypes and have often been used as the defining ligand. In this overview, the structures and therapeutic potential of conotoxins active at ion channels are highlighted. The activity and structures are then contrasted with ciguatoxins, which are responsible for the food poisoning known as ciguatera. A universal liquid chromatography/mass spectrometry approach to the detection of these classes of toxins is briefly discussed. PMID- 10688262 TI - Bites and stings from venomous animals: a global overview. AB - Venomous and poisonous animals are a significant cause of global morbidity and mortality. This Seminar will cover selected aspects of these animals, their venoms/poisons, and their clinical impact on humankind, from a global perspective, but with a distinctive Australian flavor and a clinical emphasis. Venomous snakes are found throughout most of the world, including many oceans, and have evolved a variety of highly effective toxins and methods of delivery. Their impact on humans is considerable, most current data suggesting they cause in excess of 3 million bites per year with more than 150,000 deaths. Particularly in the rural tropics, snakebite morbidity and mortality has a significant human medical and economic toll. The major groups of snakes causing bites are the vipers, the elapids (cobra type), the sea snakes, the side-fanged vipers, and the back-fanged colubrids. Australian venomous snakes are nearly all elapids and have evolved some of the most toxic of all snake venoms. Their effects include potent procoagulants and anticoagulants, neurotoxins, myotoxins, and nephrotoxins, but a distinct absence of the major local necrotoxins found in some non-Australian elapids and many vipers. The effect of these toxins on humans is not limited to envenoming, for the toxins are proving invaluable as research tools and diagnostic agents, and may even have a future as precursors of therapeutic agents. Because of the high toxicity and diversity of Australian elapids, a variety of monovalent antivenoms have been developed. There is also a venom detection kit to determine the type of snake and allow targeted antivenom therapy. The kit has also increased information available on diagnostic patterns of envenoming for each species. Australia is also home to the world's most lethal spiders, the funnel webs of eastern Australia, as well as the red back spider, the single most common reason for antivenom treatment in Australia. The latter spiders have been accidently exported to Japan. Within the marine environment exist a vast array of toxic animals, both poisonous and venomous, which not only cause morbidity and mortality in humans, but offer an incredibly rich array of valuable toxins. Australian waters contain some of the most lethal and medically problematic species, presenting a diverse range of clinical problems. PMID- 10688263 TI - Toxicology and evaluation of microcystins. AB - This paper reviews the toxicity and tumor-promoting properties of microcystins. Methods for screening and/or identification of microcystins in environmental samples are discussed and compared. Specific emphasis is placed on newly developed extraction/detection methods, e.g., solid phase microextraction (SPME) technique, and capillary electrophoresis coupled with laser-induced fluorescence detection. The results of a kinetic analysis of the effects of microcystins on phosphorylase-a binding to phosphatase-2A using a surface plasmon resonance biosensor are also presented. PMID- 10688264 TI - Clinical toxicology: a tropical Australian perspective. AB - Tropical Australia has an amazing diversity of venomous fauna, from "the world's most venomous creature," the multi-tentacled (chirodropid) box jellyfish Chironex fleckeri, to aggressive spiders whose venom remains to be characterized. All genera of highly venomous Australasian elapid snakes are present, except for tiger snakes. Most notable is the taipan (Oxyuranus scutellatus), with the most efficient "snap-release" biting mechanism of any snake and venom components causing the full constellation of clinical envenoming features: coagulopathy from fibrinogen depletion (procoagulant), neurotoxicity (predominantly presynaptic neurotoxin) and rhabdomyolysis (myotoxin). Brown snakes (Pseudonaja textilis and P. nuchalis) now account for most snake bite fatalities in Australia, as a result of severe coagulopathy and a poorly defined early scenario of collapse, postulated to be caused by profound hypotension caused by transient myocardial dysfunction associated with prothrombin activation. Other venomous entities include paralyzing ticks, the blue-ringed octopus, stone fish and other marine animals with venomous spines, paralyzing cone shells, and a wide range of jellyfish including Carukia barnesi and possibly other four-tentacled (carybdeid) box jellyfish causing the Irukandji syndrome. PMID- 10688265 TI - Post-mortem drug analyses in bone and bone marrow. PMID- 10688266 TI - Capillary electrophoresis: a new analytical tool for forensic toxicologists. AB - In capillary electrophoresis, electrophoretic or electrokinetic separations are carried out in tiny capillaries at high voltages (10-30 kV), thus achieving high efficiency (N > 105), resolution power, and mass sensitivity (down to 10(-18)-10( 20) moles). The main characteristics of capillary electrophoresis are versatility of application (from inorganic ions to large DNA fragments), use of different separation modes with different selectivity, low demands on sample volume, negligible running costs, possibility of interfacing with different detection systems including mass spectrometry, and the ruggedness and simplicity of the instrumentation. Capillary electrophoresis applications in the forensic sciences are now rapidly growing, particularly in forensic toxicology. The present paper briefly describes the basic principles of capillary electrophoresis and presents a selected review of its main applications to the analysis of illicit/controlled drugs in biologic samples. An original analytical approach to the determination of carbohydrate deficient transferrin, a new marker of chronic alcohol abuse, based on capillary electrophoresis is also described. It is concluded that the peculiar separation mechanisms and the high complementarity of capillary electrophoresis to chromatography make it a new powerful tool of investigation in the hands of forensic toxicologists. PMID- 10688267 TI - Cylindrospermopsin, a cyanobacterial alkaloid: evaluation of its toxicologic activity. AB - This paper describes the natural occurrence of the toxin, cylindrospermopsin, in two species of cyanobacteria found in Australia. The structure and chemical properties of this compound are described along with a nontoxic analog of cylindrospermopsin. The results of both intraperitoneal (IP) and oral dosing of mice show that hepatotoxicity is the main effect of cylindrospermopsin in vivo, but that a thrombohemorrhagic phenomenon is observed in a proportion of dosed animals. It has been shown that the toxin can be metabolized in vivo and that a bound metabolite occurs in the liver. Cytotoxicity experiments using cell cultures show that cylindrospermopsin is more cytotoxic to isolated rat liver hepatocytes than to other cell types. Risk assessment calculations show that guideline values for cylindrospernopsin in drinking water should lie in the low microgram per liter range. PMID- 10688268 TI - Setting standards of practice in therapeutic drug monitoring and clinical toxicology: a North American view. AB - Standards for therapeutic drug monitoring (TDM) must address the factors required for a valid TDM result. These are that the test be ordered with an appropriate indication, samples be obtained at the appropriate time, analysis be precise and accurate, and the result be interpreted and used correctly. General indications for ordering drug concentrations are to assess patient compliance, lack of response, adverse effects, initial or new baseline concentration after steady state has been achieved, and drug interactions. In establishing standards for clinical toxicology testing, the following points need to be considered: relatively few drugs will be involved in the majority of overdoses in a given location, the majority of drug overdoses and poisonings are treated symptomatically and supportively, and there are a relatively small number of drugs the testing of which may provide useful information in the emergency setting. An effective toxicology screen can be designed by first developing a list of candidate drugs that have antidotes, require specific treatment, are frequently encountered, or have a delayed onset of toxicity. Once such a list is in hand, it can be further evaluated to determine whether qualitative or quantitative testing is most appropriate and if rapid, convenient methods of analysis are available. PMID- 10688269 TI - Athletic drug testing: an analyst's view of science and the law. AB - Detection of performance-enhancing drugs in sports has received increasing visibility. Athletic drug testing uses sophisticated technology and both interindividual (population) and intraindividual reference ranges to interpret data. An effective program must incorporate educational and adjudication components in addition to testing. The difficult interface between science and the law is evident in many recent sports arbitration decisions. PMID- 10688270 TI - Drugs in sports: analytical trends. AB - A minority of athletes continues to use prohibited drugs in sports to enhance performance. Athletes discovered using these drugs can be subject to severe penalties, often resulting in media and public scrutiny, especially at major events such as the Olympic Games. The International Olympic Committee (IOC) has set out the classes of the substances it bans in the IOC Medical Code. In many cases "old" drugs such as anabolic steroids are still used, and current testing regimes can test for these. Advances in the therapeutic treatment of illness have resulted in new drugs or practices, many of which are difficult to detect and which have been turned to the sinister role of performance enhancement. Detection of some newly developed drugs which have been placed on the banned list offers a major challenge to laboratories involved in sports dope testing. In some cases this requires research into new applications of research techniques. These techniques involve the novel use of gas chromatography/ mass spectrometry (GC/MS) techniques, high-resolution mass spectrometry (HRMS), carbon isotope ratio mass spectrometry, and immunoassay techniques. PMID- 10688271 TI - Advances in understanding drug metabolism and its contribution to variability in patient response. AB - Recent advances in the understanding of the molecular biology and genetic variability of human drug metabolizing enzymes, particularly cytochromes P450, have contributed immensely towards clarifying the pharmacokinetics and pharmacodynamics of many existing drugs, and are increasingly important in the development of new chemical entities. However, whereas this knowledge has implications for therapeutics, appreciation and application in clinical practice have either been limited or yet to be realized. For example, the debrisoquine polymorphism was discovered 20 years ago, but controlled prospective studies to evaluate its clinical significance and pharmacoeconomic impact are few. Come the millennium it will be seen to what extent traditional therapeutic drug monitoring, involving the measurement of specific plasma drug concentrations, may be complemented or replaced by more general phenotyping or genotyping screens for human drug metabolizing enzymes. PMID- 10688272 TI - Pharmacogenetic methods as a complement to therapeutic monitoring of antidepressants and neuroleptics. AB - A short review of the metabolism of psychoactive drugs and the pharmacogenetic factors regulating the enzymes involved is presented here. The potential clinical usefulness of phenotyping and genotyping individuals, with regard to their drug metabolic capacity, is discussed. Indications for genotyping CYP2D6 and a flow scheme for the combined use of conventional therapeutic drug monitoring (TDM) and pharmacogenetic methods for optimizing dosage-schedules of psychoactive drugs are suggested. PMID- 10688273 TI - Pharmacogenetics of classical and new antipsychotic drugs. AB - Several classical antipsychotic drugs, i.e., chlorpromazine, haloperidol, perphenazine, thioridazine and zuclopenthixol; and some new neuroleptic drugs, i.e., risperidone and sertindole, are metabolized predominantly by cytochrome P450 (CYP) 2D6. Significant relationships have been reported between the steady state plasma concentrations (Css) of some classical neuroleptics and the CYP2D6 activity or genotype. Several of these drugs also potently inhibit the CYP2D6 activity. These facts explain several drug metabolic interactions of the classical drugs. Two studies failed to show that the CYP2D6 activity predicts the therapeutic effects of haloperidol or perphenazine. Some studies have suggested that the poor metabolizer phenotype is associated with the development of oversedation during treatment with the classical drugs, but other studies have been inconsistent or negative. The CYP2D6 phenotyping and genotyping appear to be useful in predicting the Css of some classical drugs, but their usefulness in predicting clinical effects must be further explored. PMID- 10688274 TI - Therapeutic drug monitoring of antiretrovirals in human immunodeficiency virus infection. AB - The era of antiviral therapy directed against HIV-1 has now entered its second decade. In the twelve years since the FDA approved the first antiretroviral drug zidovudine there have been a number of seminal developments that have revolutionized the approach to therapy. These advances converged to change the treatment paradigm from one of therapeutic nihilism to that of cautious optimism. First, several trials demonstrated that combination therapy of nucleoside reverse transcriptase inhibitors (NRTIs) is superior to monotherapy in extending survival and delaying disease progression. Second, the concept of virologic latency in asymptomatic HIV-infected patients was revised. Mathematic modelling demonstrated that there is an ongoing high level of virus production driving a rapid turnover of CD4 cells at all stages of infection. Hence it was concluded that the aim of antiretroviral therapy (ART) should be to "hit early and hit hard." Third, significant advances in molecular virology facilitated the development of quantitative methods to measure the circulating HIV plasma RNA. HIV viral load has been shown to be a sensitive predictor of disease progression and a valuable marker of response to therapy. However, none of these developments would have translated into improved patient care without the advent of two new classes of drugs-the protease inhibitors (PIs) and the nonnucleoside reverse transcriptase inhibitors (NNRTIs). PMID- 10688275 TI - Antiepileptic drug pharmacogenetics. AB - Until recently, the drug treatment of epilepsy has been empirical. However, in recent years as a result of improved understanding of seizure neurochemistry and mechanisms of action of antiepileptic drugs (AEDs), drug treatment has become somewhat more rational. Nevertheless, it is currently impossible to predict which patient will respond to a particular AED, and which patient will experience adverse drug effects. The only practical way to determine whether a patient will find a drug useful is to try it. The discovery of genetic polymorphism in drug metabolism has contributed significantly to understanding of the variability in dose-concentration relationships, susceptibility to adverse effects, and susceptibility to seizure intractability. The discovery that predisposition to seizure intractability and expression of brain neuromolecules consequent to seizures is under genetic control may allow a more rational approach to AED choice. In the future, treatment may be guided by a series of pharmacogenetic tests, which would serve not only to choose the most appropriate AED (in terms of efficacy and adverse effects) but also to monitor the antiepileptogenic and the evolution status of the disease. PMID- 10688276 TI - Influence of dietary components on the gastrointestinal metabolism and transport of drugs. AB - There is widespread recognition that the ingestion of a meal is associated with a number of physiologic changes (gastric pH, gastric emptying, hepatic blood flow, etc.) that can significantly alter the rate and extent of drug absorption. It is also well recognized that the components of food can alter drug absorption through alterations in drug solubility. The nutritional status of a patient can also contribute to variability in the pharmacokinetics of certain drugs. The more recent finding that grapefruit juice can increase the bioavailability of certain drugs, by reducing presystemic intestinal metabolism, has led to renewed interest in the area of 'food-drug interactions.' Particular interest has focused on the effects of the grapefruit flavonoid, naringin, and the furanocoumarin, 6',7' dihydroxybergamottin, on the activity of intestinal CYP3A4. The possibility that grapefruit juice might affect drug absorption via an interaction with intestinal P-glycoprotein (P-gp) is also being explored. The growing use of herbal extracts and phytopharmaceuticals raises a new challenge-will the use of these products cause changes in the pharmacokinetics of 'conventional' drugs? As a case in point, consider the phytoestrogenic isoflavones, which are being promoted for a number of health benefits. Isoflavones such as genistein and daidzein can inhibit oxidative and conjugative metabolism in vitro and interact with transporters such as P-gp and the canalicular multispecific organic anion transporter. Given that P gp and canalicular multispecific organic anion transporter are involved in the intestinal absorption and biliary excretion of a wide range of drugs and metabolites, it is reasonable to suspect that isoflavones may alter drug disposition in humans. However, this possibility has not been explored. PMID- 10688277 TI - Role of P-glycoprotein in drug disposition. AB - P-glycoprotein (Pgp), which is coded by human MDR1 (multidrug resistance) gene, is an energy-dependent efflux pump that exports its substrates out of the cell. Human Pgp is present not only in tumor cells but also in normal tissues including the kidney, liver, small and large intestine, brain, testis, and adrenal gland, and the pregnant uterus. This tissue distribution indicates that Pgp plays a significant role in excreting xenobiotics and metabolites into urine and bile and into the intestinal lumen, and in preventing their accumulation in the brain. The roles of Pgp in drug disposition include a urinary excretion mechanism in the kidney, a biliary excretion mechanism in the liver, an absorption barrier and determinant of oral bioavailability, and the blood-brain barrier that limits the accumulation of drugs in the brain. The inhibition of the transporting function of Pgp can cause clinically significant drug interactions and can also increase the penetration of drugs into the brain and the accumulation of drugs in the brain. Digoxin is a typical substrate for Pgp, which regulates the renal tubular secretion and brain distribution of digoxin. At present, potent Pgp inhibitors are being investigated in clinical trials aimed at overcoming the intrinsic or acquired multidrug resistance of human cancers. The clinical application of these Pgp inhibitors should take into consideration the physiologic function of pgp. PMID- 10688278 TI - Evaluation of an immunoassay for mycophenolic acid. PMID- 10688279 TI - Exercise, mobility and aging. AB - The elderly population is growing both in size and in proportion of the total population. The costs to the community of the elderly being in poor health are also growing proportionately. The beneficial effects of exercise on various physiological and psychological parameters in the elderly have been well established. The effects of exercise on the mobility and independence of the elderly are also of primary concern, their maintenance being an important exercise goal. Impaired balance and gait are the 2 most significant risk factors for limited mobility and falls in the elderly. It is important to understand the effects of aging and exercise on these risk factors. PMID- 10688280 TI - Evidence and possible mechanisms of altered maximum heart rate with endurance training and tapering. AB - Exercise physiologists, coaches and athletes have traditionally used heart rate (HR) to monitor training intensity during exercise. While it is known that aerobic training decreases submaximal HR (HRsubmax) at a given absolute exercise workload, the general consensus is that maximum HR (HRmax) is relatively unaltered regardless of training status in a given population. It has not been seriously postulated as to whether HRmax can change modestly with aerobic training/detraining. Despite several sources stating that HRmax is unaltered with training, several studies report that HRmax is reduced following regular aerobic exercise by sedentary adults and endurance athletes, and can increase upon cessation of aerobic exercise. Furthermore, evidence suggests that tapering/detraining can increase HRmax. Therefore, it is plausible that some of the same mechanisms that affect both resting and HRsubmax may also play a role in altered HRmax. Some of the proposed mechanisms for changes in HRmax that may occur with aerobic training include autonomic (extrinsic) factors such as plasma volume expansion and(enhanced baroreflex function, while some nonautonomic (intrinsic) factors are alteration of the electrophysiology of the sinoatrial (SA) node and decreased beta-adrenergic receptor number and density. There is a high correlation between changes in both maximal oxygen uptake (VO2 max) and HRmax that occurs with training, tapering and detraining (r= -0.76: p < 0.0001; n = 314), which indicates that as VO2max improves with training, HRmax tends to decrease, and when detraining ensues, HRmax tends to increase. The overall effect of aerobic training and detraining on HRmax is moderate: effect sizes based on several studies were calculated to be -0.48 and +0.54, respectively. Therefore, analysis reveals that HRmax can be altered by 3 to 7% with aerobic training/detraining. However, because of a lack of research in the area of training on HRmax, the reader should remain speculative and allow for cautious interpretation until further, more thorough investigations are carried out as to the confirmation of mechanisms involved. Despite the limitations of using HR and HRmax as a guide to training intensity, the practical implications of monitoring changing HRmax are: (i) prescribed training intensities may be more precisely monitored; and (ii) prevention of overtraining may possibly be enhanced. As such, it may be sensible to monitor HRmax directly in athletes throughout the training year, perhaps at every macrocycle (3 to 6 weeks). PMID- 10688281 TI - Sports haematology. AB - While the crucial role of haemoglobin in aerobic exercise has been well accepted, there is still a great deal of controversy about the optimal haematological parameters in the athletic population. The initial part of this review will examine the question of anaemia in athletes. The most common finding in athletes is a dilutional pseudoanaemia that is caused by a plasma volume expansion, rather than an actual blood loss. It is not a pathological state and normalises with training cessation in 3 to 5 days. This entity should be distinguished from conditions associated with lowered blood counts, such as intravascular haemolysis or iron deficiency anaemia. The evaluation of true anaemia states in the athlete must take into account not only blood losses secondary to exercise, such as foot strike haemolysis or iron losses through sweat, but non-athletic causes as well. Depending on the age and sex of the athlete, consideration must be given to evaluation of the gastrointestinal or genitourinary systems for blood loss. Finally, a comprehensive nutritional history must be taken, as athletes, especially women, are frequently not consuming adequate dietary iron. The second section of the paper will deal with the very contentious issue of sickle cell trait. While there have been studies demonstrating an increased risk of sudden death in people with sickle cell trait, it is still quite rare and should not be used as a restriction to activity. Further, studies have demonstrated that patients with sickle cell trait have an exercise capacity that is probably normal or near normal. However, in the cases of sudden death, it has been secondary to rhabdomyolysis occurring among sickle cell trait athletes performing at intense exertion under hot conditions, soon after arriving at altitude. The recommendations are that athletes with sickle cell trait adhere to compliance with the general guidelines for fluid replacement and acclimatisation to hot conditions and altitude. The final section of the paper examines the issue of haematological manipulation for the purposes of ergogenic improvement. Although experiments with blood doping revealed improvements in running time to exhaustion and maximal oxygen uptake, the introduction of recombinant erythropoietin has rendered blood doping little more than a historical footnote. However, the improvements in performance are not without risk, and the use of exogenous erythropoietin has the potential for increased viscosity of the blood and thrombosis with potentially fatal results. Until a definitive test is developed for detection of exogenous erythropoietin, it will continue to be a part of elite athletics. PMID- 10688282 TI - The effect of hormone replacement therapy and exercise on cardiovascular disease risk factors in postmenopausal women. AB - Following menopause, women show an increased risk of heart disease to a level equal that of men. This elevated risk is thought to be due, at least partly, to changes in blood lipid and fibrinogen levels. The purpose of this article is to review the published research on the relationship between both exercise and hormone replacement with regards to common cardiovascular disease (CVD) risk factors and the relative importance of each. Menopause is associated with increased total serum cholesterol, triglycerides and fibrinogen, and a decrease in high density lipoprotein (HDL) cholesterol levels. The major reason for these changes following menopause is believed to be a result of fluctuations in hormonal status, primarily a deficiency in estrogen. Intervention may be justified since estrogen replacement therapy has been shown to decrease the risk of developing CVD and to have a significant impact on many of the CVD risk factors. The results vary from study to study, but generally estrogen replacement has been found to decrease total cholesterol and fibrinogen, while increasing HDL cholesterol and triglycerides. All of these changes, other than the increase in triglycerides, are seen as positive. The addition of progestogen to estrogen may negate some of the beneficial changes of estrogen, most notably the increase in HDL cholesterol levels. However, progestogen has also been reported to offset the increase in triglycerides seen with unopposed estrogen replacement. Thus, there are contradictory effects (both positive and negative) of hormone replacement on CVD risk factors in women. Regular aerobic exercise and resulting improvements in cardiorespiratory fitness have consistently been shown as preventive of CVD. This decreased CVD risk is in part because of the impact of exercise on blood lipids and fibrinogen. Increased aerobic exercise is thought to improve the risk profile, mainly through an increase in HDL cholesterol levels and decreases in triglycerides and fibrinogen. Unfortunately, the majority of research supporting the effects of exercise on CVD risk factors has been done on men. Even when research has included women, very few studies have focused on postmenopausal women. However, the research done on postmenopausal women points to a significantly improved CVD risk factor profile with regular cardiorespiratory exercise. PMID- 10688283 TI - Anterior cruciate injuries in the skeletally immature athlete: a review of treatment outcomes. AB - The documentation of anterior cruciate ligament (ACL) injuries in the skeletally immature athlete has significantly increased over the past decade, primarily due to increased awareness of these injuries within this younger athletic population. The evaluation of these injuries are similar to that in the adult population. Diagnostic studies such as plain radiographs, as well as magnetic resonance imaging, can delineate the location of the ACL failure. Physical presentation most commonly includes an acute haemarthrosis and ligamentous insufficiency. Several studies have demonstrated that the diagnostic reliability of the physical examination is poor in children, especially in patients less than 12 years old. The site of ACL failure in this adolescent population is most commonly at the tibial insertion. We recommend arthroscopic or arthroscopically assisted open reduction and internal fixation with nonabsorbable sutures for all displaced tibial eminence fractures. Mid-substance ACL failures also occur in this athletic age group. The association of meniscal injuries with these ACL failures appears to be greater than 50%. Historically, poor subjective and objective outcomes have been associated with primary and extra-articular repairs. Intra-articular reconstruction is the gold standard. The issue of placing the graft across open physeal plates is under investigation. Recent animal studies as well as human clinical series have demonstrated safety in placing soft tissue, i.e. hamstring grafts, across open growth plates without subsequent angular or leg length discrepancy. Historically, non-operatively treated ACL failures are associated with poor functional outcomes as well as a high incidence of meniscal re-injury. If the treatment of an adolescent athlete with an ACL failure is to be rehabilitation until skeletal maturity, close follow-up is essential to detect functional instability, which may prompt earlier surgical reconstruction. PMID- 10688284 TI - Foot injuries and arthroscopy in sport. AB - Arthroscopy of the foot and ankle is a valuable tool for treating athletic foot injuries. The ability to evaluate and treat injuries without an extensive open approach is very important when dealing with the athletic population, allowing a much quicker rehabilitation and return to sport. The diagnostic and therapeutic indications for foot and ankle arthroscopy have increased significantly over recent years. Techniques and instrumentation have become more advanced and more readily available to the orthopaedist. This article defines the conditions in which arthroscopy is appropriate and presents current techniques for treatment. PMID- 10688285 TI - Diagnostic and procedural coding: a panacea or the promotion of mediocrity? PMID- 10688286 TI - Custom 95 degree condylar blade plate for pediatric subtrochanteric femur fractures. AB - Subtrochanteric fractures in the older child and adolescent often are not amenable to conservative methods of treatment. The anatomic constraints of the proximal femur including the small diameter of the femoral neck and the presence of the capital femoral physis may limit the type of internal fixation used in these patients. This article presents our preliminary experience with a custom 95 degree condylar blade plate for subtrochanteric fractures in older children and adolescents. PMID- 10688287 TI - A descriptive system for lower extremity evaluation in children: data for the newborn infant. AB - An efficient and practical system for examination of the lower extremities in newborn infants and children is presented. Measurements of hip rotation and abduction are performed in a supine position. During examination, the foot is held in passive correction or induced weight-bearing position to compare the tibia, forefoot, and hindfoot alignment. Thirty-six newborn infants were examined randomly to establish standard measurements for newborns. The only statistically significant difference between male and female infants occurred in hip abduction and internal and external rotation of the hips during extension. This method of examination of the lower extremity in newborn infants is useful for routine evaluation in all children. PMID- 10688288 TI - Comparison of ender nails, dynamic hip screws, and Gamma nails in the treatment of peritrochanteric femoral fractures. AB - Three hundred seventy-six patients with peritrochanteric fractures treated over a 10-year period were reviewed. In 85 patients treated with Ender nailing, the most frequent complications were leg shortening (34), external rotation failure (21), and postoperative distal gliding of the nails (7). In 183 patients treated with dynamic hip screws or a 95 degree condylar or a 130 degree blade plate, 3 presented with infection, 5 with instability, and 4 with femoral head necrosis. In 105 patients treated with Gamma nailing, the most serious complications were infections (3), inward rotation failure (2), postoperative bleeding at the insertion site (2), and shaft fracture (1). Because of its inherent instability, Ender nailing is no longer used. The implantation technique for the dynamic hip screw is safer and simpler than the various models of the ASIF blade plate. However, the dynamic hip screw has been superseded by the Gamma nail because of its absolute stability. In the beginning, Gamma nailing was technically more demanding with a higher number of intraoperative complications. In this study, the number of malalignments did not differ significantly between the dynamic hip screw (14) and the Gamma nail (11), but was high with Ender nailing (56). PMID- 10688289 TI - Variation in hospital charges for total joint arthroplasty: an investigation of physician efficiency. AB - Total joint arthroplasty is a common procedure for which consistent, clinically satisfactory outcomes are expected. Data from 796 total joint procedures performed by 20 surgeons at one center were evaluated to identify sources of variability in costs (as measured by hospital charges) where clinical outcome is expected to remain constant. Stepwise multivariate regression characterized the contribution of six variables to hospital charges listed in order of explanatory power: postoperative length of stay, surgical time, patient preoperative morbidity, units of blood transfused, perioperative complications, and procedure type (hip or knee) accounted for 46% of variability in hospital charges (multiple R2). In a subsequent analysis, after statistical adjustment for preoperative comorbid diagnoses, the sampling distribution of mean values for surgical time, total units of blood transfused, and total hospital charges were summarized and compared among surgeons. Despite adjustment for comorbid diagnoses, substantial variation and significant differences remained between surgeons in markers of resource utilization and "surgical efficiency." These findings suggest there is substantial variability in hospital charges not attributable to patient characteristics or category of procedure--a distinct and economically significant portion of this variability is practitioner specific. PMID- 10688290 TI - Musculoskeletal trauma in tobacco farming. AB - The incidence of musculoskeletal injury sustained during tobacco farming has been poorly documented. Using the trauma registry for all farm-related injuries occurring during a 16-month period, hospital charts, radiographs, and clinic charts were reviewed to identify those patients sustaining tobacco farming related injury. Twenty-three of 24 persons who sustained a farm-related injury during the study period were injured while farming tobacco. Seventeen (74%) were injured in falls from vented tobacco-drying barns, and 18 (75%) sustained skeletal injury. Extreme heat, humidity, and poor barn design and maintenance contribute to the incidence of falling. These injuries, largely underreported, may be substantially reduced by improvements in barn design and construction. PMID- 10688291 TI - Osteochondral defects of the knee. PMID- 10688292 TI - Acrometastasis from a transitional cell carcinoma of the bladder. PMID- 10688293 TI - Bilateral delayed extensor pollicis longus rupture following bilateral undisplaced distal radial fractures. PMID- 10688294 TI - Radiologic case study. Fibrous mass compressing the posterior tibial neurovascular bundle. PMID- 10688295 TI - Electrocardiographic imaging: a noninvasive imaging modality for characterization of intramural myocardial activation. PMID- 10688296 TI - Accuracy of the LocaLisa system in catheter ablation procedures. AB - Estimation of the 3-dimensional (3D) position of ablation electrodes from fluoroscopic images is inadequate in the ablation of complex arrhythmogenic substrates. We developed a new technique for real-time 3D localization of intracardiac electrodes. Regular catheter electrodes are used as sensors for a high-frequency transthoracic electrical field, which is applied via standard skin electrodes. We investigated localization accuracy by comparing measured and true interelectrode distances between the tip and the 10th electrode of a decapolar catheter, and the tip and the 4th electrode of a quadripolar catheter during catheter ablation procedures. Long-term stability was analyzed by localization of the proximal His bundle before and after slow pathway ablation. Accuracy achieved with the 54-mm distance between the two outer electrodes of the decapolar catheters was 101% +/- 15%, 95% +/- 10%, and 97% +/- 8% in the right atrium, right ventricle, and left ventricle, respectively. During catheter ablation procedures, the measured distance between the tip and 4th electrode of the mapping catheter was 100% +/- 15% in atrial flutter, 100% +/- 12% in slow pathway ablation, and 100% +/- 14% in ablations for left ventricular tachycardia. After 2 hours, localization of the proximal His bundle was reproducible within 1.4 +/- 1.1 mm. The LocaLisa technique allows for reproducible, real-time nonfluoroscopic 3D visualization of standard mapping and ablation catheters and is sufficiently accurate for the creation of linear radiofrequency lesions. The freedom of catheter choice makes the LocaLisa system an invaluable tool in catheter mapping and ablation procedures. PMID- 10688297 TI - Noncontact mapping of cardiac arrhythmias. AB - The development of new mapping systems is beginning to overcome some of the limitations of conventional techniques by offering percutaneous deployment, simultaneous acquisitions of data, high-resolution maps, and correlation of anatomy and electrophysiology, in addition to a catheter location system. The noncontact mapping system continues to undergo development and does not completely address all requirements for an ideal mapping system. The limitations of the system include deterioration in the quality of electrogram reconstruction with increasing distance between the MEA and the endocardium, the inability of the noncontact system to identify subendocardial activation, and problems with distinguishing noise from low-amplitude diastolic electrograms. PMID- 10688298 TI - Use of a 3-dimensional electroanatomical mapping system for catheter ablation of macroreentrant right atrial tachycardia following atriotomy. AB - The purpose of this study was to utilize a 3-dimensional (3D) electroanatomical mapping system (CARTO) to characterize the reentrant circuit in macroreentrant right atrial tachycardia (AT) following right atriotomy. Right atrial mapping was performed during incessant AT in a patient who had a right atriotomy for closure of an atrial septal defect. During AT, the right atrial free wall exhibited a large contiguous area of low bipolar voltage (< or =0.5 mV, 7.3 cm in length, and 6.3 cm in width). Two discrete scars, showing no electrical potential, were identified within the large low-voltage area. A larger vertical scar (thought to be from the atriotomy) and a smaller second scar (possible inferior vena cava cannulation scar) formed a narrow channel (1.5 cm in width) between these 2 scars. Right atrial activation propagated around the large upper scar, and then propagated through the channel between the 2 scars. A single application of radiofrequency current within the channel eliminated the macroreentrant AT. In conclusion, macroreentrant AT following right atriotomy was associated with 2 discrete scars and utilized the isolated channel between the 2 scars. Ablation within the channel effectively eliminated macroreentrant AT after atriotomy and eliminated the requirement for linear ablation between one or more of the scars and the tricuspid annulus. PMID- 10688299 TI - Review of methods to predict and detect atrial fibrillation in post-cardiac surgery patients. AB - Atrial fibrillation (AF) is the most common sustained arrhythmia after cardiac surgery. Postoperative AF is known to substantially lengthen hospital stay and affect patient recovery. Identification of those at risk of developing AF after surgery and early detection of AF during recovery would be extremely helpful in effective management of these patients, including targeting prophylactic therapy to prevent AF in high-risk patients. In this communication, diagnostic methods to identify those at risk of developing AF after surgery and early identification of AF before, during, and after surgery have been reviewed. Signal-averaged P wave analysis, done before surgery, identifies patients who are likely to develop AF during recovery. When combined with low ejection fraction, signal-averaged P wave can discriminate those who develop AF from those who do not. During recovery, AF can be detected early either from a detailed analysis of atrial activity in a 10 second electrocardiogram or an analysis of R-to-R intervals from an extended rhythm strip (1 minute or longer). Analysis of the 10-second electrocardiogram includes median QRST subtraction from rhythm data and detection and analysis of atrial signals in the resulting residual. AF is detected from extended rhythm strips by using a statistical model to identify the presence of characteristic irregular patterns of R-to-R intervals. PMID- 10688300 TI - Comparison of 18-lead ECG and selected body surface potential mapping leads in determining maximally deviated ST lead and efficacy in detecting acute myocardial ischemia during coronary occlusion. AB - Kornreich identified 6 body surface potential mapping (BSPM) leads outside the standard 12-lead electrocardiographic (ECG) sites for optimal recognition of ST segment elevation (+) and depression (-) during acute ischemia in anterior, inferior, and posterior myocardial zones (A+, A-, I+, I-, P+, P-). No comparison has been made between the 6 selected BSPM leads and 18-lead ECG (12 + V3-5R + V7 9) in detecting acute myocardial ischemia during coronary occlusion. Continuous 18-lead ECG and 6 selected BSPM leads were recorded in 68 patients (77 vessels) undergoing coronary angioplasty during balloon occlusion. Ischemia was defined as ST segment deviation (deltaST) > or = 100 microV > or = 1 lead from the preinflation baseline. The 18-lead ECG was a more frequent source of the maximal deltaST lead during left anterior descending artery, right coronary artery, and left circumflex artery occlusion (71 [92%]) than the 6 selected BSPM leads (5 [7%]). The 18-lead ECG was more efficacious than the 6 selected BSPM leads for detecting acute myocardial ischemia in the group as whole. The 18-lead ECG was also more efficacious for detecting right ventricular ischemia associated with proximal right coronary artery occlusion and for detecting ST segment elevation during left circumflex artery occlusion. Our findings indicate that the 18-lead ECG is the most frequent source of maximally deviated lead and is more efficacious in detecting myocardial ischemia during balloon occlusion than the 6 selected BSPM leads. The 6 selected BSPM leads do not add information above and beyond the 12- or 18-lead ECG, and thus cannot be recommended as optimal sites for continuous ST segment monitoring of patients with acute coronary syndromes. PMID- 10688301 TI - Accuracy of the EASI 12-lead electrocardiogram compared to the standard 12-lead electrocardiogram for diagnosing multiple cardiac abnormalities. AB - This study was performed to compare a derived 12-lead electrocardiogram (ECG) using a simple 5-electrode lead configuration (EASI 12-lead) with the standard ECG for multiple cardiac diagnoses. Accurate diagnosis of arrhythmias and ischemia often require analysis of multiple (ideally, 12) ECG leads; however, continuous 12-lead monitoring is impractical in hospital settings. EASI and standard ECGs were compared in 540 patients, 426 of whom also had continuous 12 lead ST segment monitoring with both lead methods. Independent standards relative to a correct diagnosis were used whenever possible, for example, echocardiographic data for chamber enlargement-hypertrophy, and troponin levels for acute infarction. Percent agreement between the 2 methods were: cardiac rhythm, 100%; chamber enlargement-hypertrophy, 84%-99%; right and left bundle branch block, 95% and 97%, respectively; left anterior and posterior fascicular block, 97% and 99%, respectively; prior anterior and inferior infarction, 95% and 92%, respectively. There was very little variation between the 2 lead methods in cardiac interval measurements; however, there was more variation in P, QRS, and T wave axes. Of the 426 patients with ST monitoring, 138 patients had a total of 238 ST events (26, acute infarction; 62, angioplasty-induced ischemia; 150, spontaneous transient ischemia). There was 100% agreement between the 2 methods for acute infarction, 95% agreement for angioplasty-induced ischemia, and 89% agreement for transient ischemia. EASI and standard 12-lead ECGs are comparable for multiple cardiac diagnoses; however, serial ECG changes (eg, T-wave changes) should be assessed using one consistent 12-lead method. PMID- 10688302 TI - Are additional right precordial and left posterior ECG leads useful for the diagnosis of right ventricular infarct and posterior infarct? Also a plea for the revival of vectorcardiography. PMID- 10688303 TI - The extended-length electrocardiogram (XL-ECG): a new tool for predicting risk of sudden cardiac death. PMID- 10688304 TI - Paradoxical QRST integral changes with ventricular repolarization dispersion. AB - Body surface QRST integral (QRSTI) maps have been shown theoretically to reflect disparity of intrinsic repolarization properties and have been experimentally linked to increased arrhythmia susceptibility. Paradoxically, a lower magnitude of QRSTI in patients with heart disease and at risk for arrhythmias has been reported. We hypothesized that this paradoxical reduction in QRST magnitude is a consequence of increased heterogeneity of repolarization gradients in normal hearts. We generated QRSTI using a previously published heart model to compare QRSTI for aligned and random repolarization gradients. The heart model consisted of 50,000 cubic units in an anatomically correct arrangement that included parameters to simulate anisotropic conduction and inhomogeneous distribution of refractoriness. Body surface potential maps (BSPMs) were generated on a torso surface assuming a homogeneous torso and using the boundary element method for normal alignment of repolarization gradients and spatially reassigned repolarization values that randomized repolarization directions. QT duration was measured by the subtraction of Q onset time from T offset time on the BSPM. T offset was defined as the last potential to be detected at intervals of 3 ms that was above the threshold of 0.1 mV during recovery. The time of T offset showed a consistent tendency to shift to the left posterior and to split. When slow conduction velocities were assigned, BSPMs showed delayed propagation and multiple extrema. QRSTI showed systematic magnitude decrease with increasing randomness of repolarization gradient direction. Ventricular fibrillation (VF) could be induced by successive extrastimuli under the conditions of over 70% deviation and slow conduction of 0.5 m/s for the longitudinal direction. In conclusion, a possible explanation for the paradoxical reduction in QRSTI in the presence of constant repolarization disparity is the change in alignment of repolarization gradients. PMID- 10688305 TI - Detection of coronary artery disease using maximum value of ST/HR hysteresis over different number of leads. AB - We have studied the effect of the number and ordering of exercise electrocardiographic (ECG) leads when using the maximum value of the ST segment depression/heart rate (ST/HR) hysteresis over a different number of leads for the detection of coronary artery disease (CAD). The study population consisted of 127 patients with CAD and 220 patients with a low likelihood of the disease referred for an exercise test at Tampere University Hospital, Finland. The lead system used was the Mason-Likar modification of the standard 12-lead system, and exercise tests were performed on a bicycle ergometer. The number of leads was studied using lead sets consisting of first 2 leads, then 3 leads, and so on, up to all 12 leads. The criterion for the order of inclusion of the next lead in the new lead set was based on the maximized area under the receiver operating characteristic (ROC) curve for the new lead set. The importance of the number of leads was evaluated by means of three different approaches: ROC analysis; using a fixed partition criterion of 0.01 mV; and using a fixed specificity value of 80%. According to the results, the most powerful diagnostic capacity of an individual lead was in lead V5, and the most deficient diagnostic capacities were in leads aVL and V1. Using the maximum search procedure, it was possible to improve the diagnostic capacity of the ST/HR hysteresis by anything from 4 up to a maximum of 8 leads. After that it started to decrease rapidly. In conclusion, this study suggests that the diagnostic capacity of the ST/HR hysteresis could be improved by increasing the number of leads. However, the selection of leads is of major importance when using the maximum value of the ST/HR hysteresis over the leads in the detection of CAD. PMID- 10688306 TI - Reentrant arrhythmias and their control in models of mammalian cardiac tissue. AB - We use detailed biophysical and simplified models of excitation propagation in heart muscle to study the properties of reentrant arrhythmias. Using a detailed model of excitation combined with a bidomain description of propagation and action of electric current, we have obtained a theoretical estimation for the defibrillation threshold consistent with experimental data. Reentry acts as a spiral wave, propagating around a region of block, the core. A series of properly timed low-voltage stimuli can cause directed "resonant" drift of this block and act as a low-voltage defibrillation strategy. Experimentally observed activation patterns in fibrillating tissue are more complicated than the simplest spiral wave patterns. This is due to complicated geometry, the 3-dimensional nature of the tissue, and its anisotropy and inhomogeneity. However, some fibrillation patterns can be produced by a single reentrant wave, modulated by inhomogeneous tissue properties and Wenckebach frequency division. PMID- 10688307 TI - Experimental cardiac tachyarrhythmias in guinea pigs. AB - Despite years of intense research into the mechanisms of defibrillation, there remain many unanswered questions. In many fields, hypotheses are first tested in rodent models before confirming the results in larger animals. This work suggests the guinea pig as a rodent model for defibrillation. Twenty-eight guinea pigs were studied, all male retired breeders weighing over 900 g. T-wave stimuli (upper limit of vulnerability [ULV]) were given after 15 rapid pacing beats, since the rapid pacing has been suggested to extend the tachyarrhythmia. Defibrillation (DF) was attempted after 5 seconds. The correlation between the ULV50 and DF50 in guinea pigs (0.82, n = 8) is very close to that seen in dogs (0.85). Also, the sensitivity of the DF50 to waveform is similar (476 +/- 176 for monophasic vs 364 +/- 94 V for biphasic P < 0.005, n = 10). The dose-response curve widths (2.3 +/- 1.7 for ULV vs 1.9 +/- 1.8 for defibrillation, n = 10) show the same trend of increasing curve widths for ULV, and similar magnitude to dogs (mean 1.8). We rarely (<1.5%) observed spontaneous conversion in less than 10 seconds. The guinea pig can be used as a model for defibrillation as it shows many of the same characteristics as dogs. PMID- 10688308 TI - Graded response and restitution hypotheses of ventricular vulnerability to fibrillation: insights into the mechanism of initiation of fibrillation. AB - According to the upper limit of vulnerability (ULV), failed defibrillation (DF) shocks reinitiate ventricular fibrillation (VF) by falling on the vulnerable period of one or more of the fibrillation wavefronts. The failed shock first induces reentry (stage I VF), which within few cycles degenerate to stage II VF. We developed 2 hypotheses of vulnerability that explain DF failure using isolated and intact in situ ventricles. Activation maps were constructed with high resolution electrodes and action potential (AP) recorded with microelectrodes. According to the graded response (GR) hypothesis, reentry is formed when a critical shock strength induces a GR that transiently increases local refractoriness. The GR propagates and initiates distal regenerative activity that propagates around the site of block to reenter through it as it recovers. Ultrastrong shocks prevent reentry by converting unidirectional block to bidirectional block by excessive increase in refractoriness, a finding that supports the ULV hypothesis. In situ ventricle stimulus-induced termination of reentry and stage I VF (protective zone) could be explained by the GR hypothesis. The induced functional reentry with periods of 100 to 160 ms engages the steep (unstable) portion of the AP duration restitution curves (slope >1) that promotes meandering and breakup. This leads to transition from stage I to stage II VF (the restitution hypothesis). We conclude that the GR and restitution hypotheses provide an insight into the mechanism of ventricular vulnerability to fibrillation induced by a stimulus. These hypotheses provide a new paradigm for effective antifibrillatory strategies. PMID- 10688309 TI - Arrhythmogenic changes in action potential configuration in the ventricle induced by DC shocks. AB - Failure of defibrillation by direct current (DC) shocks is the result in part of new ventricular tachyarrhythmias induced by the shocks. We investigated the arrhythmogenic substrate produced by the shocks. Fluorescent action potential (AP) signals were recorded from rabbit hearts perfused in vitro with the use of our original optical recording system. Localized application of 10-ms shocks (S2) during the plateau phase of APs by basic stimuli (S1) caused field intensity (FI) dependent changes in APs: (a) S2 > 7 V/cm caused additional depolarization, giving rise to a prolongation of AP duration (APD); (b) With S2 > 20 V/cm, terminal repolarization was inhibited, and subsequent postshock S1 APs for 1 to 5 min were characterized by decreases in the maximum diastolic potential and amplitude of APs; and (c) S2 > 30 V/cm often resulted in a prolonged refractoriness, oscillation of membrane potential leading to ventricular tachycardia or fibrillation (VT/VF). The right ventricle was more susceptible than other regions for the aftereffects of high-intensity shocks. Using an 8 channel recording system, we compared the effect of 10-ms monophasic (M) and 5/5 ms biphasic (B) shocks applied to the whole ventricles with FI of 1 to 20 V/cm at the signal recording sites. B shocks were less potent than M shocks in the FI dependent action potential duration (APD) prolongation, and in the shock-induced enhancement of APD dispersion. Incidence and duration of VT/VF induced by M shocks were significantly greater than those by B shocks. These findings suggest that DC shocks will cause two types of arrhythmogenic substrate: one induced at sites of high FI, and the other at sites with moderate FI. The former would produce local block or focal repetitive excitation due to prolonged depolarization and oscillation of membrane potential, and the latter circuitous movement of wavefronts through an enhancement of spatial inhomogeneity of repolarization. PMID- 10688310 TI - Random nature of fibrillation leads to the probabilistic nature of defibrillation. PMID- 10688311 TI - Using digital versus analog ECG data in clinical trials. AB - In 54 patients, we measured 3 different electrocardiogram (ECG) parameters required in the Symphony 1 trial and compared various combinations of pairs of measurements from 4 different sets of analog data and 1 set of digital data. Particularly for measurements of the durations of R waves in lead aVF, we found poor intraobserver and interobserver reliability and poor agreement between analog and digital data. There was much better agreement in measurements of the amplitudes of S waves in lead V3. The poorer agreement involving durations appears to be due to difficulties in accurately identifying the offsets of portions of the QRS complex, especially in lead AVF. We conclude that, particularly in measurements of duration, digital and analog data are not equivalent. PMID- 10688312 TI - "Add-on" research in clinical trials: are we asking the right questions? PMID- 10688313 TI - The use of tomographic myocardial perfusion scanning to evaluate an electrocardiographic salvage estimation method in patients with acute myocardial infarction: an AMISTAD substudy. Acute Myocardial Infarction Study Adenosine. PMID- 10688314 TI - ECG subanalyses in clinical trials: an investigator's perspective. PMID- 10688315 TI - The initial electrocardiographic pattern in acute myocardial infarction: correlation with infarct size. PMID- 10688316 TI - The dynamics of vortex-like reentry wave filaments in three-dimensional computer models. AB - Recent studies using computer simulation and biological studies in 2-dimensional excitable media have suggested that spiral wave reentrant activation and its core dynamics are important elements in the mechanism of functional reentrant tachyarrhythmias, such as atrial and ventricular fibrillation. However, vortex like reentry has been observed in homogeneous 3-dimensional excitable media, and the dynamics of the related "filaments," which have 3-dimensionally connected "cores" in 2 dimensions, have not been clarified. In order to determine whether the filaments of vortex-like reentry waves can be observed in 3-dimensional media using a mathematical ionic current heart model and whether the abnormal ionic currents in myocardium affect the complexity of the filaments, we studied the qualitative features of vortex-like reentry dynamics using mathematical models in computer simulations. We employed the Luo-Rudy Phase I and the FitzHugh-Nagumo models for our heart media, which were cubic and ventricular shaped, and consisted of 8,000,000 and 5,636,654 myocardial units, respectively. Functional reentry, in the form of vortex waves, was induced in the media by the S1-S2 method. The vortex-like reentry waves and their filaments were displayed by computer graphics. Computations were performed on an NEC SX-4 supercomputer (NEC, Tokyo, Japan) using programs written in C language. Computer simulation studies have shown that the filament dynamics of vortex-like reentry in the original Luo Rudy model is considerably more complex than that in the FitzHugh-Nagumo model. However, when we mathematically modified the L-type calcium current and shortened the action potential duration, just as occurs with sustained rapid ventricular pacing, the dynamics of the vortex-like reentrant wave fronts and the filaments were similar in both models. Our results suggested that the original character of myocardium causes drastic changes in filament shape and location, resulting in intricate functional reentrant waves, and that if the L-type calcium current is depressed, the complexity of the dynamics of the filaments are decreased to some degree. PMID- 10688317 TI - Localization of intramural necrotic regions using electrocardiographic imaging. AB - Recent studies have demonstrated that electrocardiographic imaging (ECGI) is a novel noninvasive modality for exploring the spread of electrical activation within the ventricular wall. In this study, our goal was to explore the ability of ECGI in reconstructing epicardial potentials and electrograms in the ventricles damaged by localized necroses (<2 cm2). An anatomical model of the human ventricular myocardium was used to simulate activation sequences initiated at 428 epicardial and endocardial pacing sites distributed over the right ventricular and left ventricular free walls. From these realistic sequences, we simulated extracardiac potentials at epicardial (202 sites) and torso surfaces (352 sites) using boundary element model of the human torso. ECGI in terms of the L-curve was applied to compute epicardial potentials and unipolar electrograms (202 sites). Inversely computed electrograms correlated well with those simulated by an anatomical model (r > 0.9 at 68% of sites). Specifically, ECGI accurately reconstructed the following features that have been observed during measurements on the exposed canine hearts: (a) an epicardial potential pattern with a central minimum and two maxima, with the minimum positioned above the pacing site; (b) a complete transient loss of one of the positive areas in the epicardial potential pattern when the necrosis was located subepicardially; and (c) a transient gap in the expanding positive areas of the epicardial potential pattern when the necrosis was located intramurally or subendocardially. Findings of our study indicate that ECGI provides detailed reconstruction of patterns of myocardial activation in the presence of localized necroses and may be useful in the assessment of arrhythmogenic substrate in the clinical setting. PMID- 10688318 TI - Differential response of transmural dispersion of repolarization and torsade de pointes to beta-adrenergic agonists and antagonists in three models of the long QT syndrome. PMID- 10688319 TI - Noninvasive indices of repolarization and its dispersion. AB - In experimental studies using Langendorff perfused, isolated canine hearts immersed in a torso-shaped electrolytic tank we studied repolarization and its dispersion using direct epicardial measurements and newly derived, noninvasive body surface indices. Activation recovery intervals (ARIs) measured from 64 epicardial sites based on differences between activation times (ATs) and recovery times (RTs) provided direct measures of repolarization. The indirect, torso surface indices were derived from inflections of the root-mean-square (RMS) voltage of the torso tank surface electrocardiograms recorded simultaneously with the epicardial data. For cycle lengths ranging from 300 to 900 ms, and electrolyte temperatures ranging from 32 degrees C to 40 degrees C we calculated mean, variance, and range of ATs, RTs, and ARIs from the epicardium. From epicardial and torso surface RMS waveforms, we used times of R and T peaks and their differences to estimate mean ATs, RTs, and ARIs, respectively. The RMS T wave width as determined from the second derivative inflections on either side of the T peak served as an estimate of the dispersion of RTs. In parallel studies, we showed that the direct measures of repolarization and its dispersion were reflected in RMS waveforms generated from the epicardial electrograms themselves. In this study, we confirm that the torso and epicardial RMS waveforms reflect comparable information for estimating repolarization and its dispersion. Furthermore, the derived measures provide a method to assess mean ARIs and dispersion of RTs on a beat-to-beat basis and during abnormal (ectopic ventricular) activation sequences. PMID- 10688320 TI - Transmural dispersion of repolarization and arrhythmogenicity: the Brugada syndrome versus the long QT syndrome. AB - Recent studies have shown that ventricular myocardium is composed of at least 3 electrophysiologically distinct cell types: epicardial, endocardial, and M cells. Action potentials recorded from epicardial and M cells, unlike those recorded from endocardium, display a spike-and-dome morphology, the result of a prominent transient outward current-mediated phase 1. M cells are distinguished from endocardial and epicardial cells by the ability of their action potential to prolong disproportionately in response to a slowing of rate and/or to agents with class III actions. This intrinsic electrical heterogeneity contributes to the inscription of the electrocardiogram as well as to the development of a variety of cardiac arrhythmias. The transmural dispersion of repolarization is in large part responsible for the inscription of the J wave and T wave of the electrocardiogram. Because full repolarization of epicardium defines the peak of the T wave and that of the M cells, the end of the T wave, the interval between the peak and the end of the T wave provides a valuable index of transmural dispersion of repolarization. Differences in the response of the 3 cell types to pharmacologic agents and/or pathophysiological states often results in amplification of intrinsic electrical heterogeneities, thus providing a substrate as well as a trigger for the development of reentrant arrhythmias, including torsade de pointes (TdP) commonly associated with the long QT syndrome (LQTS) and the polymorphic ventricular tachycardia/fibrillation encountered in patients with the Brugada syndrome. Early repolarization of the epicardial action potential results in abnormal abbreviation of action potential duration due to an all-or none repolarization at the end of phase 1 of the epicardial action potential. The loss of the action potential dome in epicardium but not endocardium gives rise to a large dispersion of repolarization across the ventricular wall, resulting in a transmural voltage gradient that manifests in the electrocardiogram as an ST segment elevation (or idiopathic J wave). Under these conditions, heterogeneous repolarization of the epicardial action potential gives rise to phase 2 reentry, which provides an extrasystole capable of precipitating ventricular tachycardia/fibrillation (or rapid TdP). Experimental models displaying these phenomena show electrocardiographic characteristics similar to those of the Brugada syndrome as well as those encountered during acute ischemia. Transmural dispersion of repolarization is also greatly amplified in LQTS. Disproportionate prolongation of the M-cell action potential contributes to the development of long QT intervals, wide-based or notched T waves, and a large transmural dispersion of repolarization, which provides the substrate for the development of a polymorphic ventricular tachycardia closely resembling torsade de pointes. An early afterdepolarization-induced triggered beat is thought to provide the extrasystole that precipitates TdP. Pharmacologic models of the LQT1, LQT2 and LQT3 forms of LQTS mimic the distinctive electrocardiographic, electrophysiologic, and pharmacologic responses observed in patients with these 3 different genetic syndromes. In LQTS, as in the Brugada syndrome, a mutation in an ion channel gene (in some cases the same gene--SCN5A) is responsible for the development of a large transmural dispersion of repolarization, which serves to provide the arrhythmogenic substrate tha can lead to sudden death. PMID- 10688321 TI - Time-domain analysis of beat-to-beat variability of repolarization morphology in patients with ischemic cardiomyopathy. AB - There is growing evidence that beat-to-beat changes in ventricular repolarization contribute to increased vulnerability to ventricular arrhythmias. Beat-to-beat repolarization variability is usually measured in the electrocardiogram (ECG) by tracking consecutive QT or RT intervals. However, these measurements strongly depend on the accurate identification of T-wave endpoints, and they do not reflect changes in repolarization morphology. In this article, we propose a new computerized time-domain method to measure beat-to-beat variability of repolarization morphology without the need to identify T-wave endpoints. The repolarization correlation index (RCI) is computed for each beat to determine the difference between the morphology of repolarization within a heart-rate dependent repolarization window compared to a template (median) repolarization morphology. The repolarization variability index (RVI) describes the mean value of repolarization correlation in a studied ECG recording. To validate our method, we analyzed repolarization variability in 128-beat segments from Holter ECG recordings of 42 ischemic cardiomyopathy (ICM) patients compared to 36 healthy subjects. The ICM patients had significantly higher values of RVI than healthy subjects (in lead X: 0.045 +/- 0.035 vs. 0.024 +/- 0.010, respectively; P < .001); 18 (43%) ICM patients had RVI values above the 97.5th percentile of healthy subjects (>0.044). No significant correlation was found between the RVI values and the magnitude of heart rate, heart rate variability, QTc interval duration, or ejection fraction in studied ICM patients. In conclusion, our time domain method, based on computation of repolarization correlation indices for consecutive beats, provides a new approach to quantify beat-to-beat variability of repolarization morphology without the need to identify T-wave endpoints. PMID- 10688322 TI - Autonomic nerve activity and long QT interval syndrome: a role of acetylcholine and alpha-adrenoceptor. PMID- 10688323 TI - Cellular basis for long QT, transmural dispersion of repolarization, and torsade de pointes in the long QT syndrome. AB - Genetic studies have identified four forms of congenital long QT syndrome (LQTS) caused by mutations in ion channel genes located on chromosomes 3 (LQT3), 7 (LQT2), 11 (LQT1), and 21 (LQT5). Preliminary clinical studies have reported different phenotypic electrocardiographic patterns and different sensitivity to pacing or pharmacological therapy for each genotype. A transmural electrocardiogram and transmembrane action potentials from epicardial, M, and endocardial cells were simultaneously recorded from an arterially perfused wedge of canine left ventricle. Isoproterenol (100 nmol/L) in the presence of chromanol 293B (30 micromol/L), an I(Ks) blocker (LQT1 model), produced a preferential prolongation of M-cell action potential duration (APD), resulting in an increase in transmural dispersion of repolarization (TDR) and a broad-based T wave, as commonly seen in LQT1 patients. D-Sotalol (100 micromol/L), an I(Kr) blocker (LQT2 model), and ATX-II (20 nmol/L), an agent that augments late I(Na) (LQT3 model), also produced a preferential prolongation of M-cell APD, an increase in TDR, and low-amplitude T wave with a bifurcated appearance (LQT2), and late appearing T wave (LQT3), respectively. APD-, QT-, and TDR-rate relations were much steeper in the LQT3 model than in either the LQT1 or LQT2 model, whereas the rate relations in the LQT1 and LQT2 models were both steeper than those under control conditions. Spontaneous and programmed electrical stimulation-induced torsade de pointes (TdP) were observed in all 3 models. Propranolol (1 micromol/L), a beta blocker, completely prevented the effect of isoproterenol to persistently or transiently increase TDR and to induce TdP in the LQT1 and LQT2 models, but facilitated TdP in the LQT3 model. Mexiletine, a class IB Na+ channel blocker, dose-dependently (2-20 micromol/L) abbreviated the QT and APD more in the LQT3 model, but decreased TDR and suppressed TdP in the 3 models. PMID- 10688324 TI - Changes in autonomic activity and ventricular repolarization. AB - An increase in sympathetic activity, manifested by shortening of RR intervals (RRi) and changes in RRi variability, precedes and possibly triggers ventricular tachyarrhythmias (VTAs) by altering repolarization. We examined the effects of autonomic activity on the projection of repolarization as detected by body surface potential maps (BSPMs). We recorded 32 lead/192-point BSPMs during passive head-up tilt, tilt + infusion of isoproterenol, rapid atrial pacing, and atrial pacing + infusion of isoproterenol. Changes in QT; recovery time; activation-recovery interval (ARi); T-wave amplitude; and QT, QRST, and ST integrals and their dispersion were analyzed. Autonomic effects on sinus node were inferred from the Fourier transform-derived low and high frequency powers of RRi variability. Patients were divided into those with (SHD) and without structural heart disease (NSHD). Heart rate increased, whereas QT interval and ARi declined with tilt in both groups. RRi variability indices of sympathetic activity increased in NSHD but did not change in SHD. T-wave amplitudes declined in NSHD but did not change in SHD, suggesting altered responsiveness of ventricular repolarization to autonomic stimulation. Tilt and rapid atrial pacing during infusion of isoproterenol resulted in a paradoxical increase in T-wave amplitudes in some patients, similar to that observed before the onset of spontaneous arrhythmias. We conclude that altering autonomic activity by head-up tilt and/or infusion of sympathomimetic agents results in significant changes in the body surface projection of cardiac repolarization, which differ in patients with SHD from those without SHD. Similar paradoxical changes in the T-wave amplitude have been observed before the onset of spontaneous VTA, suggesting that abnormal response of repolarization to autonomic stimulation predisposes to arrhythmogenesis. PMID- 10688325 TI - Heart rate adjustment of ST depression in patients with coronary disease and negative standard exercise tests. AB - Heart rate (HR) adjustment of ST depression (STD) has been shown to correctly classify exercise test findings in up to 85% of normal subjects and patients with "equivocal" electrocardiographic (ECG) responses (> or =100 microV upsloping STD), but the performance of these methods in patients with truly negative ECG responses (<100 microV STD) has not been examined in detail. We reviewed negative standard exercise ECGs in 54 men and women (mean age 61 years) with coronary disease, comprising 16% of consecutive treadmill tests that were performed in 337 patients with angiographic coronary artery disease or stable angina. Mean STD was only 63 +/- 21 microV (0.63 mm) in these negative tests. Despite these subthreshold values for STD, the ST/HR index was abnormal (> or =1.6 microV/bpm) in 27 of 54 patients (50%) when STD was adjusted for the change in HR during exercise. Compared with patients with normal values for HR-adjusted STD, patients with an abnormal ST/HR index were slightly older (64 vs. 58 years, P < 0.05) and demonstrated a trend toward lower exercise duration (10.0 vs. 11.8 min). An abnormal ST/HR index was associated with greater subthreshold STD (73 vs. 53 microV, P < 0.0005) and smaller HR change (35 vs. 56 bpm, P < 0.0001) with exercise. Among the 27 patients with a normal ST/HR index by simple HR adjustment, 11 (44%) had abnormal ST/HR slopes (> or =2.4 microV/bpm) by the more complex linear regression method. Therefore, HR adjustment of STD contributes to the improved sensitivity of the exercise ECG by correct classification of some patients with truly negative standard tests. The magnitude of subthreshold STD and the extent of HR change with exercise both contribute to improved test performance. The increased sensitivity afforded by HR adjustment of STD highlights the importance of the precise measurement of subthreshold STD that is afforded by computerized ECG during exercise testing. PMID- 10688326 TI - ST/HR hysteresis: exercise and recovery phase ST depression/heart rate analysis of the exercise ECG. AB - ST segment depression/heart rate (ST/HR) hysteresis is a recently introduced novel computer method for integrating the exercise and recovery phase ST/HR analysis for improved detection of coronary artery disease (CAD). It is a continuous diagnostic variable that extracts the prevailing direction and average magnitude of the hysteresis in ST depression against HR during the first 3 consecutive minutes of postexercise recovery. This article reviews the development and evaluation of this new method in a clinical population of 347 patients referred for a routine bicycle exercise electrocardiographic (ECG) test at Tampere University Hospital, Finland. Of these patients, 127 had angiographically proven CAD, whereas 13 had no CAD according to angiography, 18 had no perfusion defect according to Tc-99m-sestamibi myocardial imaging and single photon emission computed tomography, and 189 were clinically normal with respect to cardiac diseases. For each patient, the values for ST/HR hysteresis, ST/HR index, end-exercise ST depression, and recovery ST depression were determined for each lead of the Mason-Likar modification of the standard 12-lead exercise ECG and maximum value from the lead system (aVL, aVR, and V1 excluded). The area under the receiver operating characteristics curve (ie, the discriminative capacity) of the ST/HR hysteresis was 89%, which was significantly larger than that of the end-exercise ST depression (76%, P < .0001), recovery ST depression (84%, P = .0063) or ST/HR index (83%, P = .0023), indicating the best diagnostic performance of the ST/HR hysteresis in detection of CAD regardless of the partition value selection. Furthermore, the superior diagnostic performance of the method was relatively insensitive to the ST segment measurement point or to the ECG lead selection. These results suggest that the ST/HR hysteresis improves the clinical utility of the exercise ECG test in detection of CAD. PMID- 10688327 TI - Exercise-induced QRS prolongation in patients with mild coronary artery disease: computer analysis of the digitized multilead ECGs. AB - Although exercise-induced QRS prolongation has been reported as a possible marker for inducible ischemia, subtleness of the prolongation makes it unidentifiable from standard, chart-recorded electrocardiograms (ECGs). To overcome such a limitation, we measured the QRS width using high-resolution ECGs and examined the diagnostic value of the exercise-induced QRS prolongation in patients before and after percutaneous transluminal coronary angioplasty (PTCA). In 16 patients with single- (n = 12) or double-vessel disease (n = 4), treadmill exercise ECG tests were performed before and after PTCA, while continuously recording 8-lead ECGs at 500 Hz. The onset of the QRS complexes was defined by the earliest deflection, and the end was defined as the latest deflection among 8 leads with the use of algebraic sum of the absolute voltage and their time derivatives (dV/dt) from all 8 leads. We compared QRS complexes before and 1 minute after exercise. Before PTCA, exercise prolonged the QRS width in all but 3 patients (unchanged in 2, decreased in 1) (84 +/- 7 to 87 +/- 8 ms, P < .005). After PTCA, it decreased in 4, was unchanged in 5, and increased in 7 (83 +/- 7 to 83 +/- 6 ms, not significant). PTCA shortened postexercise QRS width in all but 3 (unchanged in 2, increased in 1: 83 +/- 6 to 87 +/- 8 ms, P < .001). High-resolution ECGs enabled us to measure subtle QRS prolongation induced by mild ischemia. Because the QRS prolongation and ST-segment changes would reflect different aspects of myocardial ischemia, incorporating this measure into ST segment criteria might significantly improve the diagnostic accuracy for coronary artery disease. PMID- 10688328 TI - Artifact processing during exercise testing. AB - In signal processing of exercise electrocardiograms (ECGs), artifacts are a recurring problem. It is still difficult to discriminate the ECG curves from artifacts, especially in exercise ECGs and particularly in the high exercise phase. We focused on the artifact problem and worked on two new topics: the Finite Impulse Response Residual Filtering (FRF) algorithm and the Intelligent Lead Switch algorithm. The FRF algorithm reduces the baseline wander and muscle noise in the ECG stream, with much less distortion of the QRS complexes. It subtracts a continuously updated median beat from the current ECG, filters the residual signal with a high-pass and a low-pass filter, and adds the median beat to the filtered residual signal. The Intelligent Lead Switch algorithm takes advantage of the redundancy of a multilead system (eg, standard leads), which is nowadays used during exercise testing. It selects the best leads for QRS detection and thus improves the heart rate calculation, ST segment evaluation, and arrhythmia classification. PMID- 10688329 TI - The spirit of ISCE: dynamic interchange. Reflections on the 1998 ISCE meeting and progress report for 1999 and future directions. Presidential talk at Nara, Japan, 1999. International Society for Computerized Electrocardiology. PMID- 10688330 TI - Two novel anti-von Willebrand factor monoclonal antibodies. AB - Von Willebrand Factor is a multimer produced by endothelial cells and megakaryocytes, being stored in intracellular organelles, such as the Weibel Palade bodies and alpha-granules in endothelial cells and platelets, respectively. This molecule acts as a carrier protein for factor VIIIc, involved in the intrinsic pathway of blood coagulation maintaining its stability in circulation. Von Willebrand Factor also plays an important role in platelet aggregation and adhesion to injured vessel wall. It interacts with platelets through two distinct glycoproteins, GPIb and GPIIb/IIIa. We raised two monoclonal antibodies, ECA-3 and ECA-4, against human umbilical vascular endothelial cells that recognize and immunoprecipitate von Willebrand Factor. Interestingly, ECA-4 monoclonal antibody is able to completely inhibit platelet agglutination induced by ristocetin, suggesting that it binds to von Willebrand Factor close to platelet GPIb binding site. The use of monoclonal antibodies to identify von Willebrand Factor binding regions to factor VIII or platelets has been reported by others. In pulmonary hypertension, abnormalities have been detected on the multimeric structure of the molecule as well as on its proteolytic fragments, by using monoclonal antibodies. Moreover, monoclonal antibodies raised against specific regions of von Willebrand Factor molecule may allow studies of functional abnormalities of this protein in inherited and acquired disorders like subtypes of von Willebrand's disease. PMID- 10688331 TI - Influence of vortex speed on fresh versus stored platelet aggregation in the absence and presence of extracellular ATP. AB - Platelets are subjected to vastly differing shear forces under laminar and nonlaminar flow patterns throughout the tortuous cardiovascular system. Different activation pathways appear to be associated with platelet adhesion and aggregation under high shear rates vs. low shear rates. We found that platelets continue to aggregate at very low stirring rates (100 RPM) and low shear forces although significantly less than at high stirring rates (1000 RPM). These conditions may model vortices encountered in vivo, such as downstream of partially occluded blood vessels. The extent of agonist-induced platelet aggregation, at varying stir rates, remained essentially unchanged between 1200 and 600 RPM. This was true for both freshly prepared and stored platelets even though the extent of aggregation was significantly reduced with stored platelets. Agonists used were thrombin, thrombin receptor activating peptide (TRAP), SFLLRNP, the thromboxane A2 mimetic, U46619, plus epinephrine and ADP+epinephrine. At lower stir rates (100-400 RPM), little or no difference in aggregation levels was observed between fresh and stored platelets, depending upon agonist used. This may indicate that old and young platelets, in vivo, would be equally active at vessel walls exposed to blood flowing through a slow vortex at low shear rates. ATP, released from activated platelets, may act as a potent regulator of platelet aggregation within a vortex where the resident time of platelets and bioactive molecules is greater than in laminar flow regions. High levels of extracellular ATP (100 microM) inhibited agonist-induced aggregation of fresh platelets to a greater extent than stored platelets, except with ADP+epinephrine where the converse was observed. Inhibition, in general, appeared to be inversely related to stir rates. Low levels of extracellular ATP (10 nM, 1 microM) generally stimulated agonist-induced aggregations independent of stir rates and to a greater extent with stored platelets than fresh platelets. Unraveling how hemostasis functions within microenvironments may facilitate ways to further regulate this process. PMID- 10688332 TI - Defibrotide normalizes cardiovascular function hampered by established atherosclerosis in the rabbit. AB - In a previous paper we gave evidence that chronic oral defibrotide antagonizes the noxious effect of developing atherosclerosis in the cardiovascular system. In the present paper we give evidence that defibrotide is still capable of exerting beneficial effects on cardiovascular function once atherosclerosis is established. In fact, there was statistically significant amelioration by defibrotide infusion in the following, all of which were hampered by established atherosclerosis: in rabbit aorta relaxation to acetylcholine, prostaglandin E2, and 6-keto-prostaglandin F1alpha generation from rabbit aortas, rabbit heart left ventricular end-diastolic pressure, coronary perfusion pressure, and left ventricular developed pressure, vasopressor activity of acetylcholine and endothelin-1 on coronary perfusion pressure, and 6-keto-prostaglandin F1alpha generation from the rabbit heart. Since prostacyclin takes part in NO generation, is cellular protective, and inhibits 5-lipoxygenase product synthesis, its increase, caused by defibrotide, could explain defibrotide cardioprotective activity. Prostacyclin activity could be backed by prostaglandin E2, another cardioprotective prostaglandin. PMID- 10688333 TI - Kistrin inhibits human smooth muscle cell interaction with fibrin. AB - Because of the lack of function-blocking anti-integrin antibodies that react with nonprimate species, the study of the role of integrins in in vivo animal models of atherosclerosis has been limited. In contrast, peptides or small molecules have shown less species specificity and thus may be better tools to use. In an attempt to identify integrin antagonists of potential use against smooth muscle response to injury, we investigated the role of human smooth muscle cell interactions with fibrin by using a panel of integrin antagonists consisting of the snake venom disintegrin, Kistrin, as well as cyclic peptides with well defined integrin antagonists activities. We demonstrate that Kistrin, a disintegrin that inhibits beta1, beta2, beta3, and beta5 integrin interactions, had the most potent inhibitory effect. Based on our results, Kistrin or peptides with similar pan-integrin selectivity patterns are prime candidates for use as anti-integrin antagonists in further studies of atherosclerosis and restenosis. PMID- 10688334 TI - Recombinant human factor X: high yield expression and the role of furin in proteolytic maturation in vivo and in vitro. AB - Factor X/Xa plays a pivotal role in the coagulation cascade and exhibits a therapeutic potential for the treatment of factor X-deficient as well as FVIII and FIX inhibitor patients. This report describes the establishment of Chinese hamster ovary cell clones expressing recombinant human factor X up to 120 microg/mL x day and 78 microg/10(6) cells x day, that is to 100-fold higher levels than reported previously. Although propeptide removal and single chain precursor to light and heavy chain processing as well as vitamin K-dependent gamma-carboxylation became impaired at these expression levels, up to 25% of the recombinant human factor X produced was active. This represents the highest functional activity ever reported for a vitamin K-dependent protein at such an expression level. Expression of recombinant human factor X in Chinese hamster ovary cells lacking the endoprotease Furin revealed that propeptide removal still occurred, whereas single chain precursor to light/heavy chain processing was abolished. This suggests that a protease different from Furin mediates propeptide removal, a unique finding compared with the other vitamin K-dependent coagulation factors. In contrast, exposure of incompletely processed rFX molecules to soluble recombinant Furin in vitro mediated both of these cleavage reactions despite the absence of a typical argP4-xP3-lys/argP2-argP1 Furin cleavage site in the propeptide, indicating relaxed specificity in vitro. Concomitantly with the degree of processing, the functional activity of recombinant human factor X increased. Interestingly, Furin was shown to even perform correct N-terminal proteolytic trimming of FX molecules truncated amino-terminal to the P3 residue in vitro. Depending on the absence or presence of warfarin in the culture media, as well as on the processing state, four distinct recombinant human factor X light chain isoforms were observed and their structure characterized. One of these light chain forms correlated with the functional activity. Finally, the distribution of the individual light chain isoforms suggests that gamma carboxylation may be a prerequisite for propeptide removal. PMID- 10688335 TI - The high molecular mass, glycoprotein Ib-binding protein flavocetin-A induces only small platelet aggregates in vitro. AB - The direct effects of snake venom glycoprotein (GP) Ib-binding proteins on platelet receptors during the formation of platelet aggregates were determined by a particle counting method using light scattering. Flavocetin-A induces small platelet aggregates, but not medium or large ones. However, neither jararaca GPIb BP nor tokaracetin induce platelet aggregation. The flavocetin-A dose-response curve for formation of small aggregates is bell-shaped, with maximal effect at 1 to 2 microg/mL. The formation of small aggregates was not observed when fixed human platelets were used. Jararaca GPIb-BP, the anti-GPIb monoclonal antibody GUR83-35, prostaglandin I2, and ethylene diamine-N,N-dimethylformamide all inhibited flavocetin-A-induced small aggregate formation, but acetylsalicylic acid did not. Furthermore, anti-GPIIb/IIIa monoclonal antibodies, Abciximab, and YM337 significantly but partially inhibited aggregate formation, but the anti-von Willebrand factor monoclonal antibody NMC-4 had no effect. The formation of small aggregates required extracellular calcium, but flavocetin-A did not elevate cytosolic calcium. These results suggest that flavocetin-A binds to intact platelets, initiating platelet responses and inducing platelet aggregate formation by cross-linking platelets. Consequently, flavocetin-A may be a useful tool to study the mechanism of GPIb-mediated platelet activation and the structure-function relationships of GPIb. PMID- 10688336 TI - Low plasma folate in combination with the 677 C-->T methylenetetrahydrofolate reductase polymorphism is associated with increased risk of coronary artery disease in Koreans. PMID- 10688337 TI - In vitro clot lysis: a comparative study of two methods. PMID- 10688338 TI - Clinical governance and rehabilitation services. PMID- 10688339 TI - A double-blind placebo-controlled study of botulinum toxin in upper limb spasticity after stroke or head injury. AB - OBJECTIVE: To assess dose-response relationships to a single dose of botulinum toxin 'A' in upper limb spasticity associated with stroke or head injury. DESIGN: A double-blind placebo-controlled randomized dose ranging study. SETTING: A regional centre for neuroscience and a neurorehabilitation outpatient clinic. SUBJECTS: Twenty-one hemiplegic patients with troublesome upper limb spasticity. Nineteen with stroke and two with head injury. MAIN OUTCOME MEASURES: Spasticity (modified Ashworth), range of movement, posture (postural alignment and finger curl), disability (upper body dressing time and Frenchay Arm Test), patient reported global assessment scale. RESULTS: Combining data from all doses of botulinum toxin there was a significant reduction in spasticity at the wrist and fingers associated with a greater range of passive movement at the wrist and less finger curl at rest. There was a tendency for a further reduction in spasticity at elbow and wrist to occur with increasing dose but not for finger spasticity or curl. Effects present at six weeks were lost by 12 weeks except for a small improvement in elbow range of movement at the 1,500 Mu dose. There was no change in upper limb disability but a significant increase in patients' global assessment of benefit. CONCLUSION: Botulinum toxin produced beneficial effects in spasticity and passive range of movement in the hemiplegic upper limb. Increasing the dose increased the magnitude of response for impairments in some muscle groups but had little effect on duration of response. PMID- 10688340 TI - Low TENS treatment on post-stroke paretic arm: a three-year follow-up. AB - OBJECTIVE: To determine whether stroke patients with initial increases in arm motor recovery following low-frequency transcutaneous electrical nerve stimulation (low TENS) treatment go on to show long-term benefits. Also whether the same therapy results in long-term improvements in motor function, spasticity or activities of daily living (ADL). DESIGN: A three-year follow-up study. SUBJECTS: Twenty-eight stroke patients, who had participated in a randomized trial of daily treatment with low-frequency (1.7 Hz) transcutaneous electrical nerve stimulation (low TENS) on the paretic arm for three months starting 6-12 months after stroke. OUTCOMES: Fugl-Meyer Motor Performance Scale for evaluation of changes in arm motor function. A 6-point Ashworth Scale to measure spasticity. Barthel Index to evaluate performance in ADL. RESULTS: Motor function of the paretic arm had deteriorated in both treatment and control groups. Increased spasticity was seen in both groups. ADL score remained at a similar level in the low TENS group, whereas the control group had deteriorated during the same time period. CONCLUSIONS: Low TENS stimulation started 6-12 months after stroke may not have a specific effect on arm motor function years after completion of treatment. PMID- 10688341 TI - Treatment of hemiplegic shoulder pain in the Netherlands: results of a national survey. AB - OBJECTIVE: To describe the methods of treatment applied by physiotherapists, occupational therapists, rehabilitation physicians, nursing-home physicians and neurologists for hemiplegic shoulder pain, and to investigate their beliefs about the effectiveness of triamcinolone acetonide injections for this diagnosis. DESIGN: Postal questionnaire with structured and open-ended questions. If necessary, a written reminder was sent after 2-3 weeks. SUBJECTS: One hundred physiotherapists, 100 occupational therapists, 100 rehabilitation physicians, 100 nursing-home physicians and 100 neurologists in the Netherlands. These healthcare workers were all active in the rehabilitation of stroke patients. RESULTS: The response was 351 (70.2%), ranging from 58% (neurologists) to 83% (physiotherapists). Fifty-four different (combinations of) treatments were mentioned and were classified into eight treatment groups. The frequency of the first choice of treatment was: physiotherapy (32%), prevention/instruction/education (22%), oral medication (8%), local injection (7%), sling (4%), referral (3%), other therapies (4%), and different combinations (20%). In total, 86 respondents had applied local injections: 70 rehabilitation physicians, 10 nursing-home physicians and 6 neurologists. The injections used were: corticosteroids alone (51.2%), in combination with a local anaesthetic (37.2%) or a local anaesthetic only (9.3%). Belief in the effectiveness of triamcinolone injections, measured on a 0-100 point scale, was: physiotherapists median 62.5 (IQR 29.75-71.75), occupational therapists median 50.0 (IQR 43.0 63.0), rehabilitation physicians median 70.0 (IQR 56.5-80.0), nursing-home physicians median 35.0 (IQR 21.0-64.5), neurologists median 47.0 (IQR 20.0-63.0). CONCLUSIONS: As preventive measures and physiotherapy, or a combination of both, were found to be the favourite methods of treatment for hemiplegic shoulder pain in this survey, it seems that most physicians and therapists rely on a mechanical approach to hemiplegic shoulder pain. Rehabilitation physicians used additional local (anti-inflammatory) injections. PMID- 10688342 TI - Accuracy of physical and occupational therapists' early predictions of recovery after severe middle cerebral artery stroke. AB - INTRODUCTION: The ability of physical therapists (PTs) and occupational therapists (OTs) to predict level of outcome accurately was investigated prospectively in 91 severely disabled stroke patients with a first-ever middle cerebral artery (MCA) stroke. METHODS: Within the second and fifth week after stroke onset, 364 predictions were made by 59 PTs and 47 OTs about walking ability, dexterity, activities of daily living (ADL), need for additional care in ADL, time required to achieve independent walking ability and maximal level of ADL, and destination of discharge at six months after stroke. The functional recovery patterns of stroke patients were assessed by an independent observer. The accuracy of the therapists' predictions was compared with that of derived prediction models. In addition, the influence of characteristics of patients and therapists on the accuracy of the predictions was investigated. RESULTS: Compared to observed outcomes at six months after stroke, therapists' lowest accuracies of prediction were found for the moment at which maximal ADL score was achieved (rs = 0.07; p = NS), and highest accuracy was for level of dexterity of the hemiplegic arm (rs = 0.78; p <0.01). Therapists' predictions of functional outcome at six months tended to be too pessimistic. No significant differences were observed for dexterity and walking ability when the predictions by PTs and OTs were compared with those of regression models, whereas significant differences were found for the accuracies of OTs' and PTs' first prediction of destination of discharge and second predictions of outcome in ADL and need for additional care in ADL. No significant differences were found between the accuracy of PTs' and OTs' predictions, and their ability to predict functional outcome was not significantly influenced by the characteristics of patient and therapists. CONCLUSIONS: At two and five weeks after stroke, OTs and PTs can accurately predict level of walking ability and dexterity at six months. The prediction of time required for achieving maximal level of recovery, destination of discharge, outcome of ADL as well as need for additional care in ADL leaves room for improvement. PMID- 10688343 TI - A questionnaire assessment of unmet needs for rehabilitation services and resources for people with multiple sclerosis: results of a pilot survey in five European countries. Needs Task group of MARCH (Multiple Sclerosis and Rehabilitation, Care and Health Services Research in Europe). AB - OBJECTIVE: To develop an international services and needs assessment instrument (SUN) for people with multiple sclerosis and their carers and to pilot this in different countries of the European Community. DESIGN: Interview study of people with multiple sclerosis, their carers and nominated key professionals examining the unmet needs of patients and carers. SETTING: Belgium, Estonia, Greece, Italy and the United Kingdom. MAIN OUTCOME MEASURES: Needs assessment questionnaire. RESULTS: The study comprised 137 people with multiple sclerosis, 125 carers and 111 professionals. Patients reported on average 2.9 unmet needs for themselves; their carers and professionals reported on average 2.4. Needs were categorized into seven broad categories. Due to difficulties experienced by the local researchers in distinguishing between needs and objectives a large proportion of needs had to be assigned to the 'other' category. CONCLUSIONS: The SUN is a valuable and practicable tool for the identification of unmet needs for people with multiple sclerosis and their carers. Formal validation and reliability testing of the different language versions is recommended. PMID- 10688344 TI - The role of affect on the perception of disability in multiple sclerosis. AB - OBJECTIVE: To determine the prevalence of depression in multiple sclerosis in the community and to assess how the presence of depression affects patients' perception of their disability. DESIGN: Consecutive case series. SETTING: The study was carried out at a regional multiple sclerosis (MS) clinic. SUBJECTS: Eighty-eight patients with MS. MAIN OUTCOME MEASURES: Patients were asked to complete the following questionnaires: Hospital Anxiety and Depression Scale (HADS), Beck Depression Inventory (BDI), Rankin Scale of Disability/Handicap (completed by patient and physician to assess relative perceived disability) and two visual analogue scales (coping ability and perceived service adequacy). RESULTS: Thirty-nine per cent were case level for depression using the BDI criteria of Sullivan; 17% were case level for depression (34% borderline case) and 34% case level for anxiety on HADS. Depressed patients using both BDI and HADS criteria were three times more likely than nondepressed patients to perceive their disability as being greater than the physicians' perception (p < 0.001). CONCLUSION: Depression is common in MS and adversely affects patients' perception of their disability. PMID- 10688345 TI - Effect of muscle length on strength and dexterity after stroke. AB - OBJECTIVE: The effect of muscle length on strength and dexterity after stroke was investigated. The aim was to determine if poor function at a particular muscle length could be attributed solely to differential weakness at this joint angle or whether an additional problem of differential dexterity exists. DESIGN: This descriptive research study measured elbow flexor and extensor strength as well as dexterity at three elbow joint angles: 30 degrees , 60 degrees and 90 degrees flexion. Dexterity was measured independently of strength. SUBJECTS: Fifteen (seven female, eight male) chronic stroke patients (mean age 67 years) who could actively flex and extend their affected elbow participated. Ten neurologically normal control subjects (mean age 67 years) acted as controls. MAIN OUTCOME MEASURES: Strength was measured as peak elbow flexor and extensor torque at three angles; and dexterity was measured as coherence for slow and fast tracking also at three angles. RESULTS: Dexterity was not affected by muscle length but strength was and this finding was the same for both stroke and controls. While the magnitude of the torque-angle curves was not significantly different between stroke and controls, the shape of torque-angle curves was altered after stroke so that both the elbow flexors (p < 0.05) and extensors (p < 0.05) tested weaker in the testing position where they were shortest. CONCLUSION: Since there was no differential loss of dexterity, it appears that differential loss of strength, especially in the shortened range, may explain the clinical observation of poorer function at one muscle length than another after stroke. Specific training to strengthen the muscles in these ranges is therefore of clinical importance for rehabilitation. PMID- 10688346 TI - A Delphi study of self-care in a community population of people with multiple sclerosis. AB - OBJECTIVE: The aim of the study was to obtain the views and priorities of people with multiple sclerosis (MS) to inform the design of a professionally guided self care programme. DESIGN: A three-round postal Delphi survey was used as a research tool. SETTING: The study was conducted at the Centre for Research in Rehabilitation at Brunel University in London. SUBJECTS: The respondent panel consisted of 200 volunteers with MS of whom 136 responded to the survey (68%). Respondents were recruited through voluntary organizations throughout the UK. The only selection criterion was that the diagnosis of MS was confirmed by the general practitioner. MAIN OUTCOME MEASURES: The results from each of the three rounds of the Delphi survey were the outcome measures. RESULTS: One hundred and one people used ten or more self-care strategies (74%). Round 1 data revealed the diversity of practices reported, crossing many domains of life such as daily chores, leisure, relationships and physical and mental health. The top five priorities identified in rounds 2 and 3 concerned coping strategies, social support, independence in daily living, rest and mobility. Complete consensus about priorities was not achieved. However, agreement about priorities approached stability across rounds 2 and 3 and a highly significant Kendall's coefficient of concordance indicated there was good agreement within round 3 group rankings (W= 0.46, chi-squared = 499.37, df = 9, p<0.001, N= 122). CONCLUSION: Self-care practices were widespread, and those most commonly used could be identified. This survey method allows the views and priorities of this consumer group to be revealed. The information obtained can be used to develop services where the professional guides and encourages appropriate self-management based upon the issues that people with MS consider to be most important. PMID- 10688347 TI - Inter-rater reliability of the Barthel ADL index: how does a researcher compare to a nurse? AB - OBJECTIVES: To investigate whether a nonclinical research assistant, using standardized scoring criteria, can reliably administer the Barthel Activities of Daily Living (ADL) Index in a sample of elderly inpatients. DESIGN: Paired comparison of nurse and nonclinical research assistant Barthel Index assessments. SETTING: Acute hospital wards from two hospitals in a UK Healthcare Trust, with a catchment population of approximately 224,000 people. METHODS: A consecutive sample of 94 elderly patients with a variety of medical problems. MAIN OUTCOME MEASURES: Barthel ADL Index, Folstein Mini-Mental Status Examination. RESULTS: Whilst the inter-rater reliability of the Barthel Index was within acceptable boundaries, two items out of ten had only fair agreement and low crude agreement (transfer and dressing) on Cohen's kappa scores. CONCLUSIONS: Depending on the differences observed in any particular context, the Barthel Index can be applied with reasonable reliability by nonclinical staff applying the standardized scoring criteria. It should be noted, however, that the kappa coefficients between clinical and nonclinical assessors tend to be lower than those found when comparing two clinically trained assessors in previous research. PMID- 10688348 TI - Distribution of muscle strength impairments following stroke. AB - OBJECTIVE: The purpose of this study was to quantify the distribution of strength impairments soon after stroke. We were specifically interested in differences in impairments between proximal and distal actions, flexion and extension actions, and upper and lower limb actions. DESIGN: We conducted a retrospective chart review of strength scores of patients with acute stroke. SETTING: Inpatient rehabilitation unit. SUBJECTS: Forty-eight patients with a primary diagnosis of stroke underwent initial testing on admission; 31 of the same patients underwent final testing prior to discharge. MAIN OUTCOME MEASURES: The strength of eight muscle actions was assessed bilaterally using hand-held dynamometry. Force measurements obtained were expressed in newtons and as a percentage of normal. RESULTS: Strength was impaired bilaterally but more so on the side contralateral to the brain lesion. Distal muscle actions were less impaired than proximal muscle actions on the stronger side. Extension actions were less impaired than flexion actions bilaterally but primarily in the upper limbs. Upper limb actions were less impaired than lower limb actions only on the stronger side. CONCLUSIONS: With a few exceptions, our results do not support common clinical assumptions regarding the distribution of strength impairments following stroke. PMID- 10688349 TI - Ground reaction force after a sideways push as a measure of balance in recovery from stroke. AB - OBJECTIVE: To investigate if measuring ground reaction force after a sideways push at the hips gives a measure of standing balance in stroke subjects. METHODS: Fifteen control subjects and 13 right hemiparetic subjects who were able to stand independently stood with their feet on a single forceplate. Horizontal sideways pushes of 3% body weight were delivered to each side of the pelvis with the subjects held firmly in a semi-rigid belt. Measurements were made of lateral pelvic displacement (sway) and the lateral sheer component of ground reaction force (GRF). RESULTS: Right hemiparetic subjects showed significantly greater sway after a sideways push (p < 0.01) and later onset of GRF (p < 0.01) when pushed to their weak side compared with control subjects. There was also a positive correlation between sway after a sideways push and the onset latency of GRF in both strokes (0.41) and controls (0.61). The hemiparetic subjects swayed more (p < 0.01) when pushed to their weak side compared with their stronger side and their GRF latency was longer, but this latter measurement failed to reach statistical significance. No difference was seen between sides in sway or GRF latency in controls. CONCLUSIONS: The latency of GRF onset after a push at the hips in controls and in stroke subjects is related to sway and both measurements increase after a stroke. This test offers a method of measuring balance after a stroke, and serial testing of an individual after a stroke may prove a useful measure of an individual's recovery of balance. PMID- 10688350 TI - Relationship between timed 'up and go' and gait time in an elderly orthopaedic rehabilitation population. AB - OBJECTIVE: To analyse the relationship between the timed 'up and go' (TUG) and gait time in an elderly orthopaedic population, in order to determine whether additional useful information is obtained by measuring both. DESIGN: Observational study. SETTING: Jewish Rehabilitation Hospital, Laval, Quebec. SUBJECTS: Seventy-nine consecutive inpatients in the orthopaedic programme with a primary admitting diagnosis of either total hip replacement (THR), total knee replacement (TKR), or hip fracture repair. OUTCOME MEASURES: Timed 'up and go' and time to walk 10 metres. RESULTS: The admission correlation between gait time and TUG was r= 0.745. The correlation at discharge (r= 0.816) was higher than that seen on admission. The relationship between gait time and TUG was linear both at admission and discharge. The correlation between gait time and TUG was strong for patients with TKR at admission (r= 0.868) and discharge (r= 0.878), and for patients with THR, both at admission (r= 0.809) and discharge (r= 0.879). However, the correlation on admission was weaker for patients with hip fracture (r= 0.497). For slow walkers (people with a gait speed below 0.5 m/s) on admission, the correlation was moderate (r= 0.649). However, for those with gait speeds faster than or equal to 0.5 m/s, the correlation was weaker (r= 0.484). This discrepancy was no longer evident on discharge. Likewise, for patients with a fast TUG score (< 30 seconds) on admission, there was virtually no relationship between TUG and gait time (r= 0.084), although a good correlation was present for those with 'up and go' times longer than 30 seconds (r = 0.634). As with gait speed, this difference disappeared by discharge. CONCLUSIONS: The relationship between gait time and TUG in an elderly orthopaedic rehabilitation population is good, and its strength varies by specific diagnosis, mobility, and time point in the course of therapy. The two measures are not redundant in this population. PMID- 10688351 TI - An optimization of the Waterlow score using regression and artificial neural networks. AB - OBJECTIVES: To optimize the ability of the Waterlow Scale to predict individuals vulnerable to developing pressure ulcers. DESIGN: Prospective cohort study. SETTING: Two acute care UK National Health Service (NHS) providers. SUBJECTS: Four hundred and twenty-two inpatients across five specialities (general medicine, general surgery, orthopaedics, oncology and rehabilitation). INTERVENTIONS: Waterlow scores recorded weekly for 14 days post admission to hospital. MAIN OUTCOME MEASURE: Development of a pressure ulcer. RESULTS: Nonlinear analysis using neural networks did not outperform linear methods. Only five items out of 11 in the Waterlow Scale appeared to have any classification ability in this patient population. CONCLUSIONS: The Waterlow score when modelled as a linear equation appears as effective as more complicated nonlinear mappings using neural networks. Only a subset of the variables of the Waterlow Scale have predictive value in this patient population, but this is a different subset to those found in a previous study of a different client group (wheelchair users). PMID- 10688352 TI - Case-mix in rehabilitation: FIM-based function-related groups. PMID- 10688353 TI - Casemix in rehabilitation: the debate continues. PMID- 10688354 TI - Case-mix in rehabilitation: a useful way to achieve a specific goal. PMID- 10688355 TI - A new twist in trypanosome RNA metabolism: cis-splicing of pre-mRNA. AB - It has been known for almost a decade and a half that in trypanosomes all mRNAs are trans-spliced by addition to the 5' end of the spliced leader (SL) sequence. During the same time period the conviction developed that classical cis-splicing introns are not present in the trypanosome genome and that the trypanosome gene arrangement is highly compact with small intergenic regions separating one gene from the next. We have now discovered that these tenets are no longer true. Poly(A) polymerase (PAP) genes in Trypanosoma brucei and Trypanosoma cruzi are split by intervening sequences of 653 and 302 nt, respectively. The intervening sequences occur at identical positions in both organisms and obey the GT/AG rule of cis-splicing introns. PAP mRNAs are trans-spliced at the very 5' end as well as internally at the 3' splice site of the intervening sequence. Interestingly, 11 nucleotide positions past the actual 5' splice site are conserved between the T. bruceiand T. cruzi introns. Point mutations in these conserved positions, as well as in the AG dinucleotide of the 3' splice site, abolish intron removal in vivo. Our results, together with the recent discovery of cis-splicing introns in Euglena gracilis, suggest that both trans- and cis-splicing are ancient acquisitions of the eukaryotic cell. PMID- 10688356 TI - Stem-loop 1 of the U1 snRNP plays a critical role in the suppression of HIV-1 polyadenylation. AB - The inactivity or occlusion of the HIV-1 poly(A) signal when in the 5' long terminal repeat (LTR) has been mechanistically investigated. First we show that neither the homologous HIV-1 promoter nor the close proximity of this RNA processing signal to the transcript initiation site is required for the occlusion effect. Instead we demonstrate that the major splice donor (MSD) site positioned about 200 bp downstream maintains the poly(A) site in an inactive state. Although mutation of MSD results in activation of the 5' LTR poly(A) signal, this effect can be suppressed by targeting U1 snRNAs near to the mutated MSD by base pairing. We show that hybrid U7-U1 snRNAs can also suppress the poly(A) signal and that this suppression is dependent on the U1 stem-loop 1. In particular the binding site for the U1 snRNP protein 70K that binds to the loop structure of stem-loop 1 is associated with poly(A) site occlusion. These experiments were carried out with an HIV-1 proviral construct and as such emphasize the physiological importance of this splice donor-poly(A) site interaction. PMID- 10688357 TI - Position-dependent inhibition of the cleavage step of pre-mRNA 3'-end processing by U1 snRNP. AB - The 3' ends of most eukaryotic pre-mRNAs are generated by 3' endonucleolytic cleavage and subsequent polyadenylation. 3'-end formation can be influenced positively or negatively by various factors. In particular, U1 snRNP acts as an inhibitor when bound to a 5' splice site located either upstream of the 3'-end formation signals of bovine papilloma virus (BPV) late transcripts or downstream of the 3'-end processing signals in the 5' LTR of the HIV-1 provirus. Previous work showed that in BPV it is not the first step, 3' cleavage, that is affected by U1 snRNP, but rather the second step, polyadenylation, that is inhibited. Since in HIV-1 the biological requirement is to produce transcripts that read through the 5' LTR cleavage site rather than being cleaved there, this mechanism seemed unlikely to apply. The obvious difference between the two examples was the relative orientation of the 3'-end formation signals and the U1 snRNP-binding site. In vitro assays were therefore used to assess the effect of U1 snRNP bound at various locations relative to a cleavage/polyadenylation site on the 3' cleavage reaction. U1 snRNP was found to inhibit cleavage when bound to a 5' splice site downstream of the cleavage/polyadenylation site, as in the HIV-1 LTR. U1 snRNP binding at this location was shown not to affect the recruitment of multiple cleavage/polyadenylation factors to the cleavage substrate, indicating that inhibition is unlikely to be due to steric hindrance. Interactions between U1A, U1 70K, and poly(A) polymerase, which mediate the effect of U1 snRNP on polyadenylation of other pre-mRNAs, were shown not to be required for cleavage inhibition. Therefore, U1 snRNP bound to a 5' splice site can inhibit cleavage and polyadenylation in two mechanistically different ways depending on whether the 5' splice site is located upstream or downstream of the cleavage site. PMID- 10688358 TI - Mg2+-independent hairpin ribozyme catalysis in hydrated RNA films. AB - The hairpin ribozyme catalyzes RNA cleavage in partially hydrated RNA films in the absence of added divalent cations. This reaction exhibits the characteristics associated with the RNA cleavage reaction observed under standard conditions in solution. Catalysis is a site-specific intramolecular transesterification reaction, requires the 2'-hydroxyl group of substrate nucleotide A(-1), and generates 2',3'-cyclic phosphate and 5'-hydroxyl termini. Mutations in both ribozyme and substrate abolish catalysis in hydrated films. The reaction is accelerated by cations that may enhance binding, conformational stability, and catalytic activity, and is inhibited by Tb3+. The reaction has an apparent temperature optimum of 4 degrees C. At this temperature, cleavage is slow (k(obs): 2 d(-1)) and progressive, with accumulation of cleavage products to an extent of 40%. The use of synthetic RNAs, chelators, and analysis of all reaction components by inductively coupled plasma-optical spectrophotometry (ICPOES) effectively rules out the possibility of contaminating divalent metals in the reactions. Catalysis is minimal under conditions of extreme dehydration, indicating that the reaction requires hydration of RNA by atmospheric water. Our results provide a further caution for those studying the biochemical activity of ribozymes in vitro and in cells, as unanticipated catalysis could occur during RNA manipulation and lead to misinterpretation of data. PMID- 10688359 TI - Metal ion catalysis during the exon-ligation step of nuclear pre-mRNA splicing: extending the parallels between the spliceosome and group II introns. AB - Mechanistic analyses of nuclear pre-mRNA splicing by the spliceosome and group II intron self-splicing provide insight into both the catalytic strategies of splicing and the evolutionary relationships between the different splicing systems. We previously showed that 3'-sulfur substitution at the 3' splice site of a nuclear pre-mRNA has no effect on splicing. We now report that 3'-sulfur substitution at the 3' splice site of a nuclear pre-mRNA causes a switch in metal specificity when the second step of splicing is monitored using a bimolecular exon-ligation assay. This suggests that the spliceosome uses a catalytic metal ion to stabilize the 3'-oxyanion leaving group during the second step of splicing, as shown previously for the first step. The lack of a metal-specificity switch under cis splicing conditions indicates that a rate-limiting conformational change between the two steps of splicing may mask the subsequent chemical step and the metal-specificity switch. As the group II intron, a true ribozyme, uses identical catalytic strategies for splicing, our results strengthen the argument that the spliceosome is an RNA catalyst that shares a common molecular ancestor with group II introns. PMID- 10688360 TI - A tertiary interaction detected in a human U2-U6 snRNA complex assembled in vitro resembles a genetically proven interaction in yeast. AB - U2 and U6 small nuclear RNAs are thought to play critical roles in pre-mRNA splicing catalysis. Genetic evidence suggests they form an extensively base paired structure within the spliceosome that is required for catalysis. Especially in light of significant similarities with group II self-splicing introns, we wished to investigate whether the purified RNAs might by themselves be able to form a complex similar to that which appears to exist in the spliceosome. To this end, we synthesized and purified large segments of human U2 and U6 snRNAs. Upon annealing, the two RNAs efficiently formed a stable and apparently extensively base-paired (Tm = 50-60 degrees C in the presence of 20 mM Mg2+) complex. To investigate possible tertiary interactions, we subjected the annealed complex to UV irradiation, and two crosslinked species were identified and characterized. The major one links the second G in the highly conserved and critical ACAGAGA sequence in U6 with an A in U2 just 5' to U2-U6 helix Ia and opposite the invariant AGC in U6. Remarkably, this crosslink indicates a tertiary interaction essentially identical to one detected previously by genetic covariation in yeast. Together our results suggest that purified U2 and U6 snRNAs can anneal and fold to form a structure resembling that likely to exist in the catalytically active spliceosome. PMID- 10688361 TI - Calculation of the relative geometry of tRNAs in the ribosome from directed hydroxyl-radical probing data. AB - The many interactions of tRNA with the ribosome are fundamental to protein synthesis. During the peptidyl transferase reaction, the acceptor ends of the aminoacyl and peptidyl tRNAs must be in close proximity to allow peptide bond formation, and their respective anticodons must base pair simultaneously with adjacent trinucleotide codons on the mRNA. The two tRNAs in this state can be arranged in two nonequivalent general configurations called the R and S orientations, many versions of which have been proposed for the geometry of tRNAs in the ribosome. Here, we report the combined use of computational analysis and tethered hydroxyl-radical probing to constrain their arrangement. We used Fe(II) tethered to the 5' end of anticodon stem-loop analogs (ASLs) of tRNA and to the 5' end of deacylated tRNA(Phe) to generate hydroxyl radicals that probe proximal positions in the backbone of adjacent tRNAs in the 70S ribosome. We inferred probe-target distances from the resulting RNA strand cleavage intensities and used these to calculate the mutual arrangement of A-site and P-site tRNAs in the ribosome, using three different structure estimation algorithms. The two tRNAs are constrained to the S configuration with an angle of about 45 degrees between the respective planes of the molecules. The terminal phosphates of 3'CCA are separated by 23 A when using the tRNA crystal conformations, and the anticodon arms of the two tRNAs are sufficiently close to interact with adjacent codons in mRNA. PMID- 10688362 TI - tRNA-guanine transglycosylase from Escherichia coli: recognition of noncognate cognate chimeric tRNA and discovery of a novel recognition site within the TpsiC arm of tRNA(Phe). AB - tRNA-guanine transglycosylase (TGT) is a key enzyme involved in the posttranscriptional modification of tRNA across the three kingdoms of life. In eukaryotes and eubacteria, TGT is involved in the introduction of queuine into the anticodon of the cognate tRNAs. In archaebacteria, TGT is responsible for the introduction of archaeosine into the D-loop of the appropriate tRNAs. The tRNA recognition patterns for the eubacterial (Escherichia coli) TGT have been studied. These studies are all consistent with a restricted recognition motif involving a U-G-U sequence in a seven-base loop at the end of a helix. While attempting to investigate the potential of negative recognition elements in noncognate tRNAs via the use of chimeric tRNAs, we have discovered a second recognition site for the E. coli TGT in the TpsiC arm of in vitro-transcribed yeast tRNA(Phe). Kinetic analyses of synthetic mutant oligoribonucleotides corresponding to the TpsiC arm of the yeast tRNA(Phe) indicate that the specific site of TGT action is G53 (within a U-G-U sequence at the transition of the TpsiC stem into the loop). Posttranscriptional base modifications in tRNA(Phe) block recognition by TGT, most likely due to a stabilization of the tRNA structure such that G53 is inaccessible to TGT. These results demonstrate that TGT can recognize the U-G-U sequence within a structural context that is different than the canonical U-G-U in the anticodon loop of tRNA(Asp). Although it is unclear if this second recognition site is physiologically relevant, this does suggest that other RNA species could serve as substrates for TGT in vivo. PMID- 10688363 TI - Deciphering the cellular pathway for transport of poly(A)-binding protein II. AB - Poly(A)-binding protein II (PABP2) is an abundant nuclear protein that binds with high affinity to nascent poly(A) tails, stimulating their extension and controlling their length. In the cytoplasm, a distinct protein (PABP1) binds to poly(A) tails and participates in mRNA translation and stability. How cytoplasmic PABP1 substitutes for nuclear PABP2 is still unknown. Here we report that PABP2 shuttles back and forth between nucleus and cytoplasm by a carrier-mediated mechanism. A potential novel type of nuclear localization signal exists at the C terminus of the protein, a domain that is highly enriched in methylated arginines. PABP2 binds directly to transportin in a RanGTP-sensitive manner, suggesting an involvement of this transport receptor in mediating import of the protein into the nucleus. Although PABP2 is small enough to diffuse passively through the nuclear pores, protein fusion experiments reveal the existence of a facilitated export pathway. Accordingly, no transport of PABP2 to the cytoplasm occurs at 4 degrees C. In contrast, export of PABP2 continues in the absence of transcription, indicating that transport to the cytoplasm is independent of mRNA traffic. Thus, rather than leaving the nucleus as a passive passenger of mRNAs, the data suggest that PABP2 interacts with the nuclear export machinery and may therefore contribute to mRNA transport. PMID- 10688364 TI - A phylogenetic analysis reveals an unusual sequence conservation within introns involved in RNA editing. AB - Adenosine deaminases that act on RNA (ADARs) are RNA editing enzymes that convert adenosines to inosines within cellular and viral RNAs. Certain glutamate receptor (gluR) pre-mRNAs are substrates for the enzymes in vivo. For example, at the R/G editing site of gluR-B, -C, and -D RNAs, ADARs change an arginine codon (AGA) to a glycine codon (IGA) so that two protein isoforms can be synthesized from a single encoded mRNA; the highly related gluR-A sequence is not edited at this site. To gain insight into what features of an RNA substrate are important for accurate and efficient editing by an ADAR, we performed a phylogenetic analysis of sequences required for editing at the R/G site. We observed highly conserved sequences that were shared by gluR-B, -C, and -D, but absent from gluR-A. Surprisingly, in contrast to results obtained in phylogenetic analyses of tRNA and rRNA, it was the bases in paired, helical regions whose identity was conserved, whereas bases in nonhelical regions varied, but maintained their nonhelical state. We speculate this pattern in part reflects constraints imposed by ADAR's unique specificity and gained support for our hypotheses with mutagenesis studies. Unexpectedly, we observed that some of the gluR introns were conserved beyond the sequences required for editing. The approximately 600-nt intron 13 of gluR-C was particularly remarkable, showing >94% nucleotide identity between human and chicken, organisms estimated to have diverged 310 million years ago. PMID- 10688365 TI - Structure of the RNA inside the vesicular stomatitis virus nucleocapsid. AB - The structure of the viral RNA (vRNA) inside intact nucleocapsids of vesicular stomatitis virus was studied by chemical probing experiments. Most of the Watson Crick positions of the nucleotide bases of vRNA in intact virus and in nucleoprotein (N)-RNA template were accessible to the chemical probes and the phosphates were protected. This suggests that the nucleoprotein binds to the sugar-phosphate backbone of the RNA and leaves the Watson-Crick positions free for the transcription and replication activities of the viral RNA-dependent RNA polymerase. The same architecture has been proposed for the influenza virus nucleocapsids. However, about 5% of the nucleotide bases were found to be relatively nonreactive towards the chemical probes and some bases were hyperreactive. The pattern of reactivities was the same for RNA inside virus and for RNA in N-RNA template that was purified over a CsCl gradient and which had more than 94% of the polymerase and phosphoprotein molecules removed. All reactivities were more or less equal on naked vRNA. This suggests that the variations in reactivity towards the chemical probes are caused by the presence of the nucleoprotein. PMID- 10688366 TI - The leader of the HIV-1 RNA genome forms a compactly folded tertiary structure. AB - The untranslated leader of the RNA genome of the human immunodeficiency virus type 1 (HIV-1) encodes multiple signals that regulate distinct steps of the viral replication cycle. The RNA secondary structure of several replicative signals in the HIV-1 leader is critical for function. Well-known examples include the TAR hairpin that forms the binding site for the viral Tat trans-activator protein and the DIS hairpin that is important for dimerization and subsequent packaging of the viral RNA into virion particles. In this study, we present evidence for the formation of a tertiary structure by the complete HIV-1 leader RNA. This conformer was recognized as a fast-migrating band on nondenaturing polyacrylamide gels, and such a migration effect is generally attributed to differences in compactness. Both the 5' and 3' domains of the 335-nt HIV-1 leader RNA are required for the formation of the compact RNA structure, and the presence of several putative interaction domains was revealed by an extensive analysis of the denaturing effect of antisense DNA oligonucleotides. The buffer conditions and sequence requirements for conformer formation are strikingly different from that of the RNA-dimerization reaction. In particular, the conformer was destabilized in the presence of Mg2+ ions and by the viral nucleocapsid (NC) protein. The presence of a stable RNA structure in the HIV-1 leader was also apparent when this RNA was used as template for reverse transcription, which yielded massive stops ahead of the structured leader domain. Formation of the conformer is a reversible event, suggesting that the HIV-1 leader is a dynamic molecule. The putative biological function of this conformational polymorphism as molecular RNA switch in the HIV-1 replication cycle is discussed. PMID- 10688368 TI - Role of neutrophils in induction of acute inflammation in T-cell-mediated immune dermatosis, psoriasis: a neutrophil-associated inflammation-boosting loop. AB - A growing body of evidence has indicated that T-cell-mediated immunity plays an important role in triggering and maintenance of psoriatic lesions. In this review we present our own experimental results as well as those from the literature related to the pathomechanism of the development of inflammatory changes in psoriatic lesions. First of all it is important to acknowledge the fact that psoriatic lesions are not uniform as assumed by many authors but that they are actually rather heterogeneous both clinically and histologically even within the same plaques. Lymphokines produced by activated T cells in psoriatic lesions have a strong influence on the proliferation of the epidermis, whose stimulated kertinocytes released several cytokines, which in turn enhance the activation state of T cells. Thus, they form a vicious cycle, a T-cell-mediated inflammation sustaining loop. Although the interaction between T-cell-mediated immunity and epidermal keratinocytes may well explain the maintenance of background "chronic" inflammatory changes diffusely observed throughout psoriatic lesions, it is not enough to explain the island-like, "acute" inflammatory changes observed within and at the border of the plaque lesions. Characteristic neutrophil accumulation under the stratum corneum can be observed in the highly inflamed and therapeutically recalcitrant areas of psoriatic lesions. They are chemotactically attracted and activated there by synergistic action of chemokines, IL-8 and Gro-a released by the stimulated keratinocytes, and particularly C5a/C5a des arg produced via the alternative complement pathway activation possibly on the surface of corneocytes. In this review, we emphasize that the accumulation of neurophils is not simply a passive event. We think that those stimulated neutrophils are able to influence not only the growth and differentiation of epidermal keratinocytes but also the activation-state of T cells by aberrant expression of HLA-DR on their surfaces as well as by their effects. These T cells in turn influence the transepidermal neutrophil migration through the effect of their lymphokines on the keratinocyte production of pro-inflammatory mediators including C3. Therefore, we propose a neutrophil-associated inflammation-boosting loop that may well explain the localized "acute" inflammatory changes scattered over the "chronic" psoriatic plaques as well as in the acutely inflamed lesions of pustular psoriasis. PMID- 10688367 TI - Mapping posttranscriptional modifications in 5S ribosomal RNA by MALDI mass spectrometry. AB - We present a method to screen RNA for posttranscriptional modifications based on Matrix Assisted Laser Desorption/Ionization mass spectrometry (MALDI-MS). After the RNA is digested to completion with a nucleotide-specific RNase, the fragments are analyzed by mass spectrometry. A comparison of the observed mass data with the data predicted from the gene sequence identifies fragments harboring modified nucleotides. Fragments larger than dinucleotides were valuable for the identification of posttranscriptional modifications. A more refined mapping of RNA modifications can be obtained by using two RNases in parallel combined with further fragmentation by Post Source Decay (PSD). This approach allows fast and sensitive screening of a purified RNA for posttranscriptional modification, and has been applied on 5S rRNA from two thermophilic microorganisms, the bacterium Bacillus stearothermophilus and the archaeon Sulfolobus acidocaldarius, as well as the halophile archaea Halobacterium halobium and Haloarcula marismortui. One S. acidocaldarius posttranscriptional modification was identified and was further characterized by PSD as a methylation of cytidine32. The modified C is located in a region that is clearly conserved with respect to both sequence and position in B. stearothermophilus and H. halobium and to some degree also in H. marismortui. However, no analogous modification was identified in the latter three organisms. We further find that the 5' end of H. halobium 5S rRNA is dephosphorylated, in contrast to the other 5S rRNA species investigated. The method additionally gives an immediate indication of whether the expected RNA sequence is in agreement with the observed fragment masses. Discrepancies with two of the published 5S rRNA sequences were identified and are reported here. PMID- 10688369 TI - Hot spot mutations in keratin 2e suggest a correlation between genotype and phenotype in patients with ichthyosis bullosa of Siemens. AB - Ichthyosis bullosa of Siemens (IBS) is a rare disorder of cornification characterized by blister formation in the upper suprabasal layers of the epidermis. Molecular analysis of IBS has identified mutations in the keratin 2e (K2e) gene, which is located in the type II keratin gene cluster on chromosome 12q. We have studied two IBS families and have identified heterozygous point mutations in codon 493 of the K2e gene in both families. Whereas a non conservative amino acid substitution at position 117 of the 2B region of K2e (E117K) was associated with a severe phenotype in family 1, family 2 showed mild clinical features as a result of a conservative substitution (E117D). These data suggest a phenotype-genotype correlation in these families. PMID- 10688370 TI - Identification of a novel mutation in keratin 1 in a family with epidermolytic hyperkeratosis. AB - Epidermolytic hyperkeratosis (EHK) is a hereditary skin disorder typified by blistering due to cytolysis. One in 100,000 individuals is affected by this autosomal-dominant disease. The onset of the disease phenotype is typically at birth. Histological and ultrastructural examination of the epidermis shows a thickened stratum corneum and tonofilament clumping around the nucleus of suprabasal keratinocytes. Linkage studies localized the disease genes on chromosomes 12q and 17q which contain the type II and type I keratin gene clusters. Recently, several point mutations in the genes encoding the suprabasal keratins, K1 and K10, have been reported in EHK patients. We have investigated a large kindred affected by EHK and identified a new point mutation in the 2B region of keratin 1 (I107T), resulting from a T to C transition in codon 478. PMID- 10688371 TI - A three-dimensional skin culture model for mouse keratinocytes: application to transgenic mouse keratinocytes. AB - The study of mouse epidermal biology has been hampered by the lack of a good in vitro model for the culture of mouse keratinocytes which allowed the reconstruction of a fully differentiated epidermis. We adapted the Prunieras' model, also called the Dead de-Epidermized Dermis model (DED), to mouse keratinocytes and showed that a neo-epidermis can be reconstructed exhibiting a complete differentiation program. We also used this model to culture transgenic mouse keratinocytes. We observed that transgene expression occurred in the correct location and that the neo-epidermis mimed previous in vivo observations obtained with integrin skin-targeted transgenic mice. Therefore, this model will be a powerful tool to further investigate normal mouse and transgenic keratinocyte biology. PMID- 10688372 TI - Langerhans cell migration is modulated by N-sulfated glucosamine moieties in heparin. AB - Dendritic cell (DC) migration into and out of tissues is important for the generation of primary immune responses to antigens encountered in tissues. In order to study the mechanisms involved in DC migration we used a skin explant system and quantitated the number of Langerhans cells (LC), which are immature precursors of DC in skin-draining lymph nodes, remaining in the epidermis in response to incubation with various biomolecules. This paper shows that LC trafficking in epidermis is a metabolically active process that is modulated by heparin, specifically by N-sulfated glucosamine moieties in heparin. This is the first demonstration of structural specificity in the biochemical requirements for DC migration in a tissue and therefore is important to understanding DC migration in general. PMID- 10688373 TI - Fibroblasts surrounding melanoma express elevated levels of matrix metalloproteinase-1 (MMP-1) and intercellular adhesion molecule-1 (ICAM-1) in vitro. AB - Tumour growth and metastasis involve the degradation of extracellular matrix components by matrix degrading enzymes produced by tumour cells and stromal fibroblasts. In this study, fibroblasts were obtained from biopsies on the border (TB) and 1 cm distant from the melanoma (TD) and cultured separately. Similar studies were performed with fibroblasts surrounding melanocytic nevi as control. The expression of matrix metalloproteinase-1 (MMP-1) mRNA and tissue matrix metalloproteinase inhibitor 1 (TIMP-1) were studied by Northern blot analysis. The activation antigen intercellular adhesion molecule-1 (ICAM-1) in TB-and TD fibroblasts was investigated by flow cytometry. In melanoma, TB-fibroblasts showed an increased expression of MMP-1 mRNA mainly in fibroblasts obtained from tumours with extended invasive growth demonstrated by Clark level whereas the expression of the major specific inhibitor TIMP-1 was unaltered. In contrast, fibroblasts surrounding benign melanocytic nevi did not express elevated levels of MMP-1. The upregulation of MMP-1 in TB-fibroblasts compared to TD-fibroblasts was maintained during cultivation. Furthermore, MMP-1 mRNA expression and MMP-1 total protein amount in normal fibroblasts were increased by melanoma cell conditioned medium. We demonstrated an increased expression of ICAM-1 in TB fibroblasts compared to TD-fibroblasts in vitro depending on the amount of inflammatory infiltrate in situ. The differences of ICAM expression disappeared during continued cell culture. These results support the idea that fibroblasts surrounding melanoma are activated and are possibly involved in the degradation of matrix proteins surrounding the tumour. PMID- 10688374 TI - Substance P induction of murine keratinocyte PAM 212 interleukin 1 production is mediated by the neurokinin 2 receptor (NK-2R). AB - The neurological system plays an important role in modulating some inflammatory skin diseases. Neuro-cutaneous interactions may be mediated by the release of neuropeptides such as substance P (SP) which activate immunocompetent cells in the skin by binding to high affinity neurokinin receptors (NKR). Since epidermal keratinocytes produce a variety of cytokines and are intimately associated with cutaneous sensory fibers, we tested the ability of these cells to participate in the cutaneous neuroimmune system by the secretion of potent cytokines such as interleukin 1 (IL-1) in response to released SP. RT-PCR studies demonstrated that cultured PAM 212 murine keratinocytes expressed mRNA for NK-2R but not NK-1R. Correspondingly, the addition of SP to these cells resulted in a rapid increase in intracellular Ca2+ levels that could be specifically blocked by an NK-2R antagonist. NK-2R was also shown in normal mouse epidermis by immunohistochemistry. SP augmented the expression of PAM 212 keratinocyte IL 1alpha mRNA in a dose and time dependent manner and this induction was inhibited by an NK-2R antagonist. Secretion of bioactive IL-1alpha by the PAM 212 keratinocytes was likewise stimulated by SP in a dose dependent manner. These data support the hypothesis that SP released from cutaneous sensory nerves contributes to neuroimmune inflammatory responses in the skin by modulating the expression and release of cytokines from epidermal keratinocytes. PMID- 10688375 TI - Proliferation and effects of UVA irradiation in cultured fibroblasts from lesions in cutaneous lupus erythematosus. AB - Cutaneous lupus erythematosus (LE) often presents clinically as chronic cutaneous lesions, healing with scar formation, and acute cutaneous lesions that are seen in systemic and subacute LE and heal without scarring. UV-light plays a role in the pathogenesis of the skin lesions, but the pathomechanism is still unclear. The aim of this study was to compare fibroblast proliferation and response to UV light by cultured fibroblasts from scarring and non-scarring LE lesions. Fibroblasts were cultured from skin lesions from 5 patients with classic discoid LE, 5 patients with subacute cutaneous LE and healthy, age-matched donors. Proliferation rate was assessed by cell counts at days 3, 6 and 9. The fibroblast cultures were irradiated with UVA and the supernatants were analysed for IL-6, TGF-beta, IL-4, soluble ICAM-1 and soluble VCAM-1. Fibroblast cultures from scarring lesions showed significantly lower cell-counts at days 3 (P = 0.01) and 9 (P = 0.009), than cultures from nonscarring lesions or controls. There were no significant differences in levels of IL-6 or TGF-beta in supernatants of irradiated fibroblasts from patients compared to controls and IL-4 and the soluble forms of ICAM-1 and VCAM-1 were below detection level. The response to UV irradiation was similar to that of normal cells in the parameters studied. In summary, cultured fibroblasts from scarring LE lesions displayed significantly decreased proliferation rates compared to non-scarring LE lesions and controls. This may be secondary to inflammatory factors, or due to a functional defect in LE fibroblasts. PMID- 10688376 TI - Immunohistochemical and molecular characterization of cultured keratinocytes after dispase-mediated detachment from the growth substratum. AB - Keratinocyte activation comprises changes in protein and gene expression pattern resulting in phenotypic and functional changes necessary for re-epithelialization such as the expression of urokinase-type plasminogen activator (uPA) and its cell surface receptor (uPA-R; CD87). As uPA and uPA-R are rapidly induced after dispase-mediated detachment of cultured normal human epidermal keratinocytes (NHEK) we hypothesized that dispase-mediated detachment may cause a similar "activation" of keratinocytes with uPA and uPA-R being only one aspect of a complex "activation reaction". To test this hypothesis we have comparatively analysed adherent versus detached keratinocyte sheets for selected indicators of keratinocyte activation by immunohistochemistry. Furthermore we have identified genes via subtraction cloning which are up-regulated upon dispase-induced detachment. The analyses provided evidence for an increased transcriptional and translational activity in detached keratinocytes, as indicated by over-expression of several ribosomal components (L3 and S10 ribosomal protein) and transcription factors (initiation factor 4A, elongation factor 1alpha). Increased proliferative activity was indicated by increased expression of the proliferation markers Ki67, keratin 6 and keratin 17. Finally, several markers of keratinocyte activation such as the integrin chain alpha(v), psoriasin, glutathion-S-transferase and heparin-binding epidermal growth factor-like growth factor were up-regulated. Furthermore mevalonate kinase, a molecule as yet unknown to be expressed in keratinocytes, was identified. The findings provide evidence that dispase mediated detachment in cultured keratinocytes induces a reaction, which comprises the up-regulation of a complex array of proliferation- and migration-related molecules. The pattern of which resembles the activation reaction observed in the re-epithelializing keratinocytes in vivo. PMID- 10688377 TI - Expression of stratum corneum chymotryptic enzyme in relation to other markers of epidermal differentiation in a skin explant model. AB - The serine proteinase stratum corneum chymotryptic enzyme (SCCE) has been proposed to be involved in the degradation of intercellular cohesive structures in cornified squamous epithelia in the process of desquamation. Since SCCE is expressed late in epidermal differentiation and is found at all sites where there is a formation of cornified epithelia it also serves as a marker for terminal epidermal differentiation. Earlier studies have shown that the link between expression and the formation of cornified cells may be stronger for SCCE than for other well characterized markers of epidermal differentiation. In an attempt to further elucidate the regulation of SCCE expression we have in this study compared the expression of SCCE with the expression of keratin 10, filaggrin and involucrin in an in vitro model with skin explants cultured for various periods of time on de-epidermized dermis at the liquid-air interface. The markers were analysed by means of immunohistochemistry. We found that the expression of SCCE preceded the expression of keratin 10 and filaggrin. In contrast to involucrin, which was expressed by all suprabasal keratinocytes, SCCE was expressed only by high suprabasal cells. Our results indicate that the expression of SCCE may be regulated in a way that differs from the regulation of the expression of keratin 10, filaggrin and involucrin. PMID- 10688378 TI - Cytokine expression in primary cutaneous germinal center cell lymphomas. AB - Physiologically, B-lymphocytes are not present in the skin. Even in pathological situations they rarely occur. In contrast, primary cutaneous B-cell lymphomas (CBCL) are characterized by proliferation of B lymphocytes within the skin. This suggests the existence of a certain microenvironment supporting homing and expansion of clonal B cells. Cytokines were demonstrated to be involved in the pathogenesis of cutaneous lymphomas of T-cell origin. Cytokine expression in cutaneous B-cell lymphoma lesions, however, has not been investigated so far. Therefore, the mRNA level of several cytokines was analyzed in biopsies from 7 patients with CBCL and compared to pleomorphic T-cell lymphoma (n = 6), psoriasis (n = 9), and healthy skin (n = 7), using a competitive RT-PCR approach. An overexpression of TNF-alpha, IL-10, and IL-6 was found. Enhanced IL-8 mRNA expression was detected in 2/7 cases. The overexpression of IL-6 and IL-10 in CBCL might be of particular importance, since these cytokines are considered to support B-cell growth. Additionally, the overexpression of IL-10 may contribute to tumor progression since this immunosuppressive cytokine might be involved in downregulation of immunological tumor surveillance, in part by inhibiting type 1 cytokine formation. In fact, we did not detect IFN-gamma and IL-2 expression. Taken together, we found a cytokine pattern in CBCL lesions which might contribute to tumor B-cell growth. PMID- 10688379 TI - Computers and the pediatric surgeon: a primer. AB - Computers have become an integral part of surgical practice. To use and maintain computers effectively, the surgeon must have a basic knowledge of the inner workings of the computer. It also is helpful to understand how the systems have evolved. Medical computing started in the financial department of large hospitals. From there it expanded to clinical data systems. Coincident with the development of clinical data systems was the introduction of the IBM personal computer in 1981 and the development of the Internet. All these events led to the use of the personal computer as a communication tool. This will shape much of how we use computers in the coming millennium. The computer is made up of several component parts. The brain of the computer is the central processing unit (CPU), which performs all of the calculations in the computer. The CPU works in concert with the random access memory (RAM) and hardware peripherals to perform tasks as directed by a program. To use this increasingly complex tool effectively, the pediatric surgeon must have a basic knowledge of information systems. It is through this knowledge that information systems may be used to enhance the efficiency of pediatric surgical practice. PMID- 10688380 TI - The Internet: past, present and future. AB - Although the Worldwide Web has just blossomed this past decade, the origins of the Internet date to the late 1950s and Cold War concerns. The technological underpinning of the Internet rests with the concept of packet switching of data over dispersed routes of electronic intercommunication. Grounded in applications for the national defense, and nurtured in the domains of science within academia, the Internet of the masses has exploded with the addition of strong commercial interest and potential. Electronic mail is still the predominant application of the Internet, and the ease of widespread and near simultaneous dissemination of such communication has led to the genesis of listservers, especially appropriate as well as predominant in Medicine. One such list for pediatric surgeons is Pedsurg-L. There is an impressive evolving etiquette for those contributing to such lists. Initiatives are already well developed with public and private partnerships for the future of the Internet. PMID- 10688381 TI - Internet resources and web pages for pediatric surgeons. AB - The Internet, the largest network of connected computers, provides immediate, dynamic, and downloadable information. By re-architecturing the work place and becoming familiar with Internet resources, pediatric surgeons have anticipated the informatics capabilities of this computer-based technology creating a new vision of work and organization in such areas as patient care, teaching, and research. This review aims to highlight how Internet navigational technology can be a useful educational resource in pediatric surgery, examines web pages of interest, and defines ideas of network communication. Basic Internet resources are electronic mail, discussion groups, file transfer, and the Worldwide Web (WWW). Electronic mailing is the most useful resource extending the avenue of learning to an international audience through news or list-servers groups. Pediatric Surgery List Server, the most popular discussion group, is a constant forum for exchange of ideas, difficult cases, consensus on management, and development of our specialty. The WWW provides an all-in-one medium of text, image, sound, and video. Associations, departments, educational sites, organizations, peer-reviewed scientific journals and Medline database web pages of prime interest to pediatric surgeons have been developing at an amazing pace. Future developments of technological advance nurturing our specialty will consist of online journals, telemedicine, international chatting, computer-based training for surgical education, and centralization of cyberspace information into database search sites. PMID- 10688382 TI - Reducing the paper load: computer-based patient records. AB - Physicians burdened with increasing paper work may find relief in computer-based patient records (CPR). CPRs may aid clinicians in the areas of billing, documentation, reporting, and data retrieval. Value-added features like decision support and event monitoring facilitate patient outcome, decrease health care costs and allow improved administration. The authors discuss obstacles in the use of computers in patient care with a focus on security, confidentiality, and Y2K. PMID- 10688383 TI - Ready or not, here it comes: the legal, ethical, and clinical implications of E mail communications. AB - Electronic mail (E-mail) has the potential to enhance the professional relationship both between physician and patient, and among physicians of the same and different specialties. Despite this promise, E-mail communication in the medical environment raises important questions of legal, ethical, and professional propriety. This article discusses the potential uses of E-mail in medical practice, identifies its benefits, and analyzes the hazards that may be associated with this new and powerful technology. In doing so, the authors hope to foster an approach to E-mail use that takes advantage of its strengths while developing policies and practices that mitigate its weaknesses. PMID- 10688384 TI - Clinical information systems. AB - Clinical information systems are the computer and information systems used by health care personnel to facilitate patient care. These systems have evolved from financial systems to true patient care systems with variable levels of functionality. Early systems provided laboratory and radiology results, and modern systems now provide copies of the radiology images and decision support for therapeutic orders. The rapidly changing technological infrastructure has created barriers to implementation of the electronic medical record, while coding schemes continue to be refined to enable data access and aggregate data analysis. Further refinement of clinical information systems is required before the potential value of these systems is realized in the clinical management of patients. PMID- 10688385 TI - Telemedicine and pediatric surgery. AB - The practice of pediatric surgery has become increasingly demanding, requiring longer working hours with less reimbursement. Although manpower in this field is adequate, there are still areas that are underserved. Advances in technology offer the pediatric surgeon tools to improve efficiency of practice, cover wider areas of practice, and service underserved locations. The purpose of this report is to introduce the pediatric surgeon to telemedicine technology and its potential impact on the practice environment. The history, key components, and current applications of telemedicine are presented. The ability to integrate this technology in pediatric surgical practice holds great potential. PMID- 10688386 TI - Comparison of bacteriologic eradication of Streptococcus pneumoniae by clarithromycin and reports of increased antimicrobial resistance. AB - OBJECTIVE: To determine whether reported increases in Streptococcus pneumoniae resistance, as determined by in vitro antimicrobial susceptibility testing, correlate with the clinical efficacy of clarithromycin in treating patients with acute exacerbations of chronic bronchitis (AECB) or community-acquired pneumonia (CAP). BACKGROUND: Surveillance data on antimicrobial resistance suggest that the overall rate of S. pneumoniae resistance in vitro in the United States has increased to approximately 45% during the past decade. S. pneumoniae is showing increased resistance to penicillin, other beta-lactams, and macrolides. Despite this increased resistance, the clinical efficacy of clarithromycin does not appear to be diminished to the degree suggested by reported resistance rates. The author examined several studies of clarithromycin in patients with AECB or CAP that demonstrate S. pneumoniae eradication rates in vivo of approximately 92%. The discordance between reported increases in resistance of S. pneumoniae isolates in vitro and the eradication rate with clarithromycin in vivo is discussed in light of 5 observations. RESULTS: First, surveillance data on S. pneumoniae resistance rates to clarithromycin may be overestimated. Second, efflux mutant strains may not be clinically resistant. Third, host immune defenses play a role in treatment outcomes. Fourth, in vitro resistance may not correlate with in vivo clinical success. Finally, clarithromycin and its active metabolite, 14-OH-clarithromycin, attain high concentrations in patients. CONCLUSION: Reported increases in the prevalence of S. pneumoniae resistance do not appear to have had proportional effects on the clinical efficacy of clarithromycin in the treatment of patients with AECB or CAP caused by S. pneumoniae. PMID- 10688387 TI - Weekly administration of alendronate: rationale and plan for clinical assessment. AB - OBJECTIVE: This paper describes the rationale and supporting data for once-weekly dosing of alendronate. BACKGROUND: Alendronate sodium, a bisphosphonate that potently inhibits bone resorption, has been shown to increase bone mass and substantially reduce the incidence of osteoporotic fractures, including fractures of the hip. The standard regimen of daily administration has generally been well tolerated. However, weekly administration may provide greater convenience to patients without compromising efficacy or tolerability. The pharmacokinetics of alendronate and bone remodeling theory predict similar efficacy for weekly and daily administration if the cumulative dose is the same. Bone resorption in individual remodeling units normally proceeds for approximately 2 weeks; alendronate inhibits the rate and extent of resorption. Because the half-life of residence on bone surfaces is several weeks, weekly administration of alendronate should inhibit bone resorption to an overall extent similar to that of daily dosing, thereby producing similar effects on bone mass and strength. Animal studies demonstrate that both weekly and daily parenteral administration of alendronate effectively increase bone mass and strength, but confirmation of efficacy is needed for weekly oral dosing in humans. Although daily bisphosphonates (alendronate and risedronate) elicited esophageal irritation in a canine model of gastroesophageal reflux, weekly dosing with alendronate at a higher unit dose did not. Thus, the lower frequency of weekly dosing with a higher unit dose may actually reduce the risk of upper gastrointestinal irritation compared with daily administration of a lower dose. CONCLUSIONS: Current safety and efficacy data justify further investigation of once-weekly dosing of alendronate. Two positive-control, double-blind, randomized trials of osteoporosis treatment and prevention are currently being performed to assess the comparability of weekly, biweekly, and daily dosing of alendronate with regard to effects on bone density, safety, and tolerability. PMID- 10688388 TI - Antimicrobial therapy of acute otitis media: review of treatment recommendations. AB - OBJECTIVE: This paper reviews 3 previously published articles that provided recommendations for antimicrobial therapy of acute otitis media (AOM) and combines them to provide revised recommendations. BACKGROUND: AOM is one of the most common pediatric infections requiring a prescription for an antimicrobial agent. The optimal approach to treatment of AOM requires early, efficacious, and practical therapy. Several experts and organizations have developed recommendations for the management of AOM, but the number of these may overwhelm the busy primary care practitioner. A MEDLINE search of the pediatric and infectious disease literature on AOM treatment recommendations was used to select 3 representative, previously published articles for this review. When selecting an agent, physicians should consider in vitro activity, particularly against drug resistant Streptococcus pneumoniae; pharmacokinetics; adverse events; palatability of the suspension; and cost. In addition, physicians' clinical experience is an important determinant. CONCLUSIONS: Amoxicillin is recommended as the first-line agent to treat uncomplicated AOM. For clinical treatment failures after 3 days of amoxicillin, recommended antimicrobial agents include oral amoxicillin/clavulanate, cefuroxime axetil, cefprozil, cefpodoxime proxetil, and intramuscular (i.m.) ceftriaxone. I.m. ceftriaxone should be reserved for severe cases or patients in whom noncompliance is expected. Tympanocentesis for identification of pathogens and susceptibility to antimicrobial agents is recommended for selection of third-line agents. PMID- 10688389 TI - Tolerability and efficacy of nabumetone and naproxen in the treatment of rheumatoid arthritis. AB - OBJECTIVE: The purpose of this study was to compare the tolerability and efficacy of nabumetone and naproxen in the treatment of patients with rheumatoid arthritis (RA). The occurrence of gastrointestinal (GI) adverse events was compared. BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) have similar efficacy at equipotent doses, but the therapeutic response to various NSAIDs often differs in individual patients. METHODS: This was a 3-month, randomized, double-blind, multicenter, parallel-group study conducted in adult patients with RA. The study had 2 phases: a 3- to 14-day washout period and a 12-week treatment period. During the treatment phase, the tolerability and efficacy of nabumetone 2000 mg/d were compared with those of naproxen 1000 mg/d. The change from baseline in efficacy variables, including global assessments, number of tender or swollen joints, and pain, was evaluated. The study was sized to provide an 80% power to detect a 15% difference in the percentage improvement on the physician's global assessment (alpha = 0.05). GI safety was assessed by monitoring the occurrence of clinically important adverse GI events. RESULTS: A total of 346 RA patients at 31 US rheumatology centers were randomly assigned to treatment (173 patients per group). The study population was predominantly white (87.0%) and female (70.5%), with a mean age of 54 years. Both treatments improved the signs and symptoms of RA, with no statistically significant differences between groups for any efficacy variables. No serious GI adverse events occurred with either NSAID. The most frequent treatment-related adverse events in both groups were predominantly GI in origin, as were those that resulted in withdrawal from the study. Diarrhea with lower abdominal pain was the most common adverse event in the nabumetone group; upper abdominal pain was the most common adverse event in the naproxen group. The only significant difference between the 2 groups was a higher incidence of diarrhea (P < 0.01) in patients receiving nabumetone. CONCLUSIONS: Nabumetone 2000 mg/d was as effective as naproxen 1000 mg/d in relieving the signs and symptoms of RA. In this study, no serious GI adverse events were observed with either NSAID, but nabumetone was associated with a higher incidence of diarrhea. PMID- 10688390 TI - Efficacy of brimonidine as replacement therapy in patients with open-angle glaucoma or ocular hypertension. AB - BACKGROUND: Patients with glaucoma or ocular hypertension may have inadequately controlled intraocular pressure (IOP) or experience adverse effects with their current medication regimens. OBJECTIVE: This post hoc reanalysis determined the effectiveness and tolerability of brimonidine used as replacement therapy in a real-life clinical practice setting. METHODS: In this multicenter, open-label, observational, 2-month study, 460 patients received brimonidine 0.2% as a 1:1 replacement for another antiglaucoma medication in their current regimen. Effectiveness was assessed by calculating the mean additional reduction in IOP from the treated baseline measurement (before the switch to brimonidine) to 2 months postbaseline, and by determining physicians' opinions of treatment effectiveness. Tolerability was determined based on quality-of-life assessments and recorded adverse events. RESULTS: Overall, brimonidine replacement significantly reduced mean (+/- SEM) IOP by an additional 2.33 +/- 0.17 mm Hg (9.8% +/- 0.9%; P < 0.001). Significant additional reductions in IOP were seen when brimonidine replaced an agent used either as monotherapy or adjunctive therapy, regardless of the drug class of the agent replaced. However, particularly good hypotensive effectiveness and additional lowering of IOP were observed when brimonidine replaced certain medications, including latanoprost (12.44%; P < 0.003) and betaxolol (13.56%; P < 0.001) monotherapy, and latanoprost (16.08%; P < 0.010) adjunctive therapy. The effectiveness of brimonidine was rated as good or excellent by 92.4% of physicians. All quality-of life variables remained favorable or improved throughout the study, and brimonidine treatment was well tolerated. CONCLUSIONS: Brimonidine 0.2% used as a 1:1 replacement for monotherapy or adjunctive therapy with other antiglaucoma drugs significantly lowered IOP from that produced by previous therapy and was well tolerated. Brimonidine offers a useful treatment option in patients who require replacement therapy. PMID- 10688391 TI - Pharmacokinetics and pharmacodynamics of cefepime administered by intermittent and continuous infusion. AB - OBJECTIVE: This study assessed the pharmacokinetics and pharmacodynamics of cefepime administered by intermittent and continuous infusion against clinical isolates of Pseudomonas aeruginosa, Enterobacter cloacae, and Staphylococcus aureus. BACKGROUND: Because beta-lactam antibiotics exhibit time-dependent bactericidal activity and lack prolonged postantibiotic effects against many bacteria, the goal of therapy is to maintain serum drug concentrations above the minimum inhibitory concentration (MIC) for the relevant pathogen over most of the dosing interval. Continuous infusion is a mode of drug administration that can provide serum drug concentrations continuously above the MIC for most bacterial pathogens. METHODS: Twelve healthy volunteers were enrolled. Each received cefepime 2 g by intermittent bolus q12h and, on another day, was randomly assigned to receive 4 or 3 g administered by continuous infusion over 24 hours. RESULTS: For the intermittent regimen, the mean (+/- SD) pharmacokinetic findings were: maximum serum concentration, 112.9 +/- 21.1 microg/mL; minimum serum concentration, 1.3 +/- 0.5 microg/mL; and half-life, 2.6 +/- 0.4 hours. For the 3 and 4-g continuous infusion regimens, steady-state serum concentrations (C(SS)) were 13.9 +/- 3.8 and 20.3 +/- 3.3 microg/mL, respectively. MICs ranged from 2 to 4, 0.125 to 8, and 2 to 8 microg/mL against P. aeruginosa, E. cloacae, and S. aureus, respectively. For the intermittent regimen, serum inhibitory titers (SITs) at 24 hours were > or = 1:2 in 46% of subjects against P. aeruginosa, 48% against E. cloacae, and 2% against S. aureus. For both continuous infusion regimens, SITs for each organism were > or = 1:2 in all subjects. CONCLUSIONS: The intermittent regimen maintained serum concentrations above the MIC for P. aeruginosa and E. cloacae in > or = 92% (11/12) of subjects for > or = 70% of the dosing interval, provided the MIC was < or = 4 microg/mL. Both continuous infusion regimens provided a C(SS) above the MIC for all organisms. However, the C(SS) was > or = 4 times the MIC only if the MIC was < or = 2 microg/mL. Only the 4-g regimen provided such concentrations against isolates with an MIC of 4 microg/mL, and neither regimen provided such concentrations when the MIC was 8 microg/mL. These findings should be applied in comparative clinical studies. PMID- 10688392 TI - Penciclovir cream for the treatment of sunlight-induced herpes simplex labialis: a randomized, double-blind, placebo-controlled trial. Penciclovir Cream Herpes Labialis Study Group. AB - OBJECTIVE: The purpose of this study was to further define the therapeutic value of penciclovir cream in the treatment of sunlight-induced herpes labialis by comparing its efficacy and tolerability with those of an inactive control (purified water). METHODS: In this randomized, double-blind, placebo-controlled, parallel-group clinical trial, lesions were induced by exposure to sunlight. Treatment was self-initiated within 1 hour of development of the signs or symptoms of a recurrence. RESULTS: Healthy male and female patients (mean age, 38.3 years; range, 18 to 81 years) who had a history of sunlight-induced herpes labialis (mean of 6 recurrences in previous 12 months) applied either penciclovir cream (n = 266) or purified water (n = 275). Penciclovir cream significantly decreased the time to lesion healing (P < 0.001), with a reduction in median time of up to 2 days. The efficacy of penciclovir cream was further supported by a significant reduction in maximum lesion area (P = 0.008), a faster loss of lesion associated symptoms (P = 0.026), and significant reductions in daily assessments of pain (P < or = 0.040), itching (P < or = 0.032), burning (P < or = 0.028), and tenderness (P < or = 0.026) as moderate or severe. These effects were reinforced by the results of the daily self-assessment of lesion attributes, with significantly fewer severe/extreme assessments of lesion size (P < or = 0.003), noticeability (P < or = 0.003), amount of scab/crust (P < or = 0.003), raised/ swollen area (P < or = 0.040), soreness/tenderness (P < or = 0.043), and overall severity (P < or = 0.001) throughout the study period. CONCLUSIONS: Penciclovir cream has demonstrated efficacy for a broad range of clinically important outcomes. Significant effects on lesion area, lesion symptoms, and other lesion attributes extend the clinical efficacy of penciclovir cream beyond lesion healing. PMID- 10688393 TI - Antibiotics and Clostridium difficile diarrhea in the ambulatory care setting. AB - OBJECTIVE: The goal of this study was to determine the prevalence of Clostridium difficile diarrhea (CDD) and the risk for CDD associated with different oral antibiotics commonly used in the ambulatory care setting. METHODS: The prevalence of CDD was determined for enrollees in 4 UnitedHealth Group-affiliated health plans between January 1, 1992, and December 31, 1994. Cases were identified based on the presence of an inpatient or outpatient claim with a primary diagnosis of diarrhea, a pharmacy claim for a prescription drug used to treat CDD, or a physician or facility claim for the C. difficile toxin test, and were confirmed using full-text medical records. Within a retrospective cohort design, periods of risk for CDD were defined on the basis of duration of antibiotic therapy. To control for potential selection bias created by heterogeneous rates of C. difficile testing and to limit confounding due to multiple antibiotic exposures, we used a nested case-control design, restricting eligibility to subjects who underwent screening for C. difficile and who had been exposed to only 1 antibiotic risk period with a single antibiotic. RESULTS: The global prevalence of CDD in 358,389 ambulatory care enrollees was 12 per 100,000 person-years. In the nested case-control study, after controlling for other risk factors, 2 antibiotics demonstrated an increased association with CDD: cephalexin (odds ratio [OR] = 7.5, 95% CI = 1.8 to 34.7) and cefixime (OR = 6.4, 95% CI = 1.2 to 39.0). CONCLUSIONS: Although CDD is thought to occur primarily in hospitalized patients, it was found to be present in an ambulatory care population, but at a low frequency. In this population, it appeared to be associated with 2 cephalosporins but not with other types of antibiotics usually linked with nosocomial CDD. Because the frequency of C. difficile testing was shown to be more common with high-risk antibiotics, CDD may be underdiagnosed in the ambulatory care setting. PMID- 10688394 TI - Ongoing clinical assessment of the safety profile and efficacy of brimonidine compared with timolol: year-three results. Brimonidine Study Group II. AB - OBJECTIVE: We compared the safety profile and efficacy of brimonidine 0.2% BID with those of timolol 0.5% BID over 3 years in patients with ocular hypertension and glaucoma. METHODS: Ninety-four eligible patients from an ongoing multicenter, interventional, double-masked clinical trial were followed through year 3, 48 receiving brimonidine 0.2% and 46 receiving timolol 0.5%. Study visits occurred at months 24, 27, 30, 33, and 36. The primary efficacy variable was mean reduction from baseline intraocular pressure (IOP) at trough. Visual acuity, visual fields, and safety variables (adverse events, ocular symptoms, heart rate, blood pressure, and laboratory test results) were monitored throughout the study. RESULTS: The 2 treatment groups were well matched, with no significant differences in demographic or clinical characteristics. Both drug regimens caused significant mean reductions from baseline IOP at trough during year 3 (P < 0.001), with no significant differences between groups at any study visit. The overall mean reduction from baseline IOP at trough was 5.02 mm Hg with brimonidine and 5.57 mm Hg with timolol (P = 0.383). Brimonidine caused reductions in IOP at trough that were equivalent to those with timolol at months 30 and 36 (within the 95% CI). Visual fields were unchanged or improved in 95% of patients in both treatment groups. Both drug regimens appeared to be safe and were well tolerated. Ocular allergy occurred in 2 brimonidine-treated patients (4.2%). There were no statistically significant differences in adverse-event reports and no clinically significant effects on any ocular or systemic safety variable in either group. CONCLUSIONS: Brimonidine 0.2% BID continues to appear to be safe, well tolerated, and effective in the long-term management of ocular hypertension and glaucoma. Over 3 years, it provided sustained IOP-lowering efficacy and visual-field preservation equal to those with timolol 0.5% BID. PMID- 10688395 TI - Economic assessment of troglitazone as an adjunct to sulfonylurea therapy in the treatment of type 2 diabetes. AB - OBJECTIVE: To assess the economic efficiency of adding troglitazone to sulfonylurea therapy to improve glycemic control. BACKGROUND: Despite the high prevalence of type 2 diabetes, existing treatment strategies often fail. New oral agents give a wider segment of the population with type 2 diabetes hope of achieving near-normal blood-glucose levels. Troglitazone, a novel chemical entity, is one promising new agent. METHODS: We conducted an economic analysis based on glycemic-control data from a randomized clinical trial comparing troglitazone with placebo, each added to glyburide. A patient simulation model was used to translate these data to long-term outcomes associated with diabetes. Patients had poorly controlled type 2 diabetes mellitus despite glyburide therapy. Risk functions of developing and progressing through nephropathy, retinopathy, neuropathy, hypoglycemia, and macrovascular disease were developed from the Diabetes Control and Complications Trial and large epidemiologic studies. Cost estimates were based on data from 5 states, all payor databases, surveys, and literature. The main outcomes of the model were cost-consequences, number of patients developing each type of complication, mean time to development of the complication, cost per life-year gained (LYG), and cost per quality adjusted life-year. RESULTS: The model predicts that for every 1000 patients treated with troglitazone, the improved glycemic control could mean that 95 to 140 fewer patients would experience one of the most severe diabetic complications (eg, blindness, end-stage renal disease, amputation), which may increase life expectancy by 2.0 years. These benefits are obtained at an additional $2100 per LYG (undiscounted). The ratio remains <$50,000 per LYG for most variations in input. CONCLUSIONS: The clinical trial demonstrated that troglitazone + glyburide improves glycemic control compared with glyburide alone. Based on these results, the model estimates fewer diabetic complications at a cost well below accepted cost-effective thresholds. PMID- 10688396 TI - Health-related quality of life and functional status of patients with rheumatoid arthritis randomly assigned to receive etanercept or placebo. AB - OBJECTIVE: To compare the functional status and well-being of patients with rheumatoid arthritis (RA) who were randomly assigned to receive placebo, etanercept 10 mg, or etanercept 25 mg during a 26-week, phase III, double-blind clinical trial. BACKGROUND: No single indicator of disease activity, severity, or therapeutic efficacy has been established for RA. During the past decade, health related quality of life, a multidimensional way to assess physical, emotional, and social aspects of a disease or its treatment, has become an important outcome in RA studies and in assessments of RA drug therapies. METHODS: A total of 234 patients completed the Health Assessment Questionnaire (HAQ), the Short-Form 36 (SF-36) (n = 48 patients), items assessing energy and mental health from the Medical Outcomes Study (MOS), and a single-item rating scale assessing current health (feeling thermometer) at baseline and several times during 6 months. RESULTS: Significant improvements from baseline to last assessment were reported with etanercept versus placebo and in the HAQ Disability Index score (ie, the total HAQ score) and all 8 HAQ categories (P < 0.05), with the exception of grip. Significant improvements with etanercept in the MOS energy and mental health subscales, current health (from the feeling thermometer), and mental and physical function components of the SF-36 were reported (P < 0.05). CONCLUSIONS: Patients receiving 10- or 25-mg doses of etanercept reported significantly better functional status and well-being than did patients receiving placebo. PMID- 10688397 TI - Savings in direct medical costs from the use of tobramycin solution for inhalation in patients with cystic fibrosis. AB - OBJECTIVE: Two identical 24-week, double-blind, placebo-controlled trials of tobramycin solution for inhalation (TOBI [PathoGenesis Corporation, Seattle, Washington]) in cystic fibrosis patients with chronic Pseudomonas aeruginosa infection were conducted in the United States. The aim of the present study was to extrapolate the US trial data to a Canadian setting, using Canadian costs to estimate the savings in direct medical costs that might result from use of a similar 24-week TOBI regimen versus usual care in 2 Canadian provinces. BACKGROUND: Cystic fibrosis is a genetic disease in which persistent respiratory infection, usually due to P. aeruginosa infection, is the major cause of morbidity and mortality. METHODS: The US trials demonstrated that TOBI produced significant improvements in pulmonary function test results, reduced sputum levels of P. aeruginosa, and resulted in a 26% reduction in the probability of hospitalization (95% CI, 2%-43% vs placebo in the clinical trials). Individual patient data from the US trials were used to calculate the mean number of days in hospital as well as the mean number of days of home intravenous or oral antibiotic therapy. To adjust for Canadian pricing, pertinent economic data were obtained from Statistics Canada and the Ontario and Quebec health ministries. Demographic and baseline data were obtained from health surveys conducted by the Canadian Cystic Fibrosis Foundation. RESULTS: Economic analysis showed that the use of TOBI for 24 weeks would result in estimated mean per-patient savings in direct medical costs (in Canadian dollars) of $4055 in Ontario and $4916 in Quebec, which would substantially offset the Canadian acquisition price of $8602 for the same 24-week period. CONCLUSIONS: Assuming that the percentage of reduction in hospital days observed in the US trials would also occur in the Canadian clinical setting, use of TOBI would reduce the use of health care services, particularly hospital days, and lead to substantial savings in direct medical costs that would offset its acquisition price. Whether this reduction actually occurs after TOBI enters the Canadian market is a subject for future investigation. PMID- 10688398 TI - An overview of salt absorption by the nephron. AB - The purpose of this brief commentary is to provide a description of how renal physiology, and more particularly, renal tubular physiology, has evolved over the past thirty years, since the occasion, obviously, is a celebration of the Thirtieth Course on Advances in Nephrology and Dialysis. My arguments will begin by quoting from Homer Smith's book, The Kidney, and merging Smith's observations, which were integrative in the sense that they did not specify detail, with detailed incursions into tubular physiology, first at a cellular level and then at a molecular level. For convenience, the nephron is divided into four functional segments: the proximal nephron; the ascending limb; the distal nephron; and the collecting duct. Each of these carries out a specific function. The proximal nephron absorbs about two-thirds of filtered sodium, without dissociating salt and water absorption. The thick ascending limb absorbs 25% of filtered Na+, but no water. The distal nephron absorbs 10% of filtered Na+ in close relation with K+ and, to some extent, H+ secretion. Finally, the collecting duct includes three kinds of cells: the cortical collecting duct, which is responsible not only for Na+ absorption and K+ secretion, but also for the bulk of the absorption of free water; the outer medullary collecting duct (OMCD), which is largely responsible for the final steep drop in urine pH which occurs between cortex and papilla; and the inner medullary collecting duct (IMCD), whose major function is the final absorption of approximately 5% of filtered Na+. PMID- 10688399 TI - Glomerular normalcy and pathosis: a "fin de millenaire" perspective. PMID- 10688400 TI - Lupus nephritis: an historical perspective 1968-1998. AB - Lupus is now regarded as a syndrome which results from several related auto immune processes, although the exact mechanisms of how the disease arises in susceptible individuals remain obscure. When the San Carlo meetings began in 1968, much less was known about the pathogenesis of autoimmunity, but the presence of autoantibodies and autoreactive cells had been worked out, the patterns of the disease and its clinical and histological expression in the kidney were well described. At that time, however, the prognosis for severe forms of lupus nephritis was miserable, although patients either milder disease might survive for decades. During the late 1950s and 1960s relatively effective palliative treatment first with corticosteroids and then cytotoxic drugs were introduced, which disappointingly remain the principal treatments 40 years later, although they have improved the prognosis of severe lupus nephritis to equal that of milder forms. However, better understanding of the immune reaction promises newer forms of more precisely-targeted treatment for the near future. PMID- 10688401 TI - Polycystic kidney disease: <<30 ans apres>>. AB - Major progress has been achieved in autosomal dominant polycystic kidney disease in the last 30 years; Progress in imaging procedures has been decisive for diagnosis (by ultasonography), management of kidney and liver complications (by CT scan), and investigation and sometimes management of intracranial aneurysms (by MRI-angiography and endovascular treatment procedures). On the other hand, progress in molecular genetics has led to the identification of PKD1 and PDK2 genes, and their respective gene products, polycystin 1 and 2. A two-hit model for cyst formation has recently been put forward. The link between the gene defects and cyst fluid formation and progression is still unknown. In addition, cystic and non-cystic lesions coexist in the disease, underlining that the primary molecular defect is located upstream of the mechanism of cyst formation. PMID- 10688402 TI - Demographic and organizational developments of maintenance dialysis therapies- past, present, outlook into the future. PMID- 10688403 TI - Mechanisms of progression of chronic renal damage. AB - The amount of protein in the urine is a strong predictor of subsequent loss of renal function. Proteinuria and tubular atrophy have been linked with progressive renal insufficiency. In the last few years several studies have indicated that smoking is also a risk factor in the progression of renal disease. In addition, a number of studies have suggested that higher levels of blood pressure are associated with a faster decline in renal function. A number of cytokines, vasoactive compounds, chemoattractant molecules and growth factors are upregulated during the course of progressive renal disease in experimental animals. Recent data indicate that vasoconstrictor substances have a key role in the initial phases of this process. In particular, angiotensin II is increased following the development of renal injury. Angiotensin in turn upregulates the expression of other factors including: transforming growth factor beta, tumor necrosis alpha, nuclear factor kappaB and several chemoattractant compounds. Other vasoactive compounds (endothelin, thromboxane A2 and prostaglandins) may also be upregulated during the course of progressive renal disease PMID- 10688404 TI - Uremic bone disease: advances over the last 30 years. PMID- 10688405 TI - The role of technology in hemodialysis. AB - The evolution of hemodialysis therapy has been characterized over the years by the search for reliable devices and supplies, for more efficient treatments and finally for a more tolerable therapy in long term dialysis patients. In this view, three steps can be identified: a) the first step was the creation of safe and reliable vascular access, dialyzers and machines. This step led to the birth of modern dialysis and treatment personalization was the logical consequence. Each patient is a single entity and he requires a specific therapy prescription and delivery. From this concept the search for adequacy and better outcomes has been generated, with the inevitable consequence that newer techniques were explored in the attempt to perform a more efficient and clinically tolerated dialysis therapy. b) The second step was the attempt to consider the intratreatment variations as possible source for dialytic morbidity. In this view, efforts were made to pre-set ultrafiltration and dialysate sodium profiles in the machine to counterbalance the negative effects of uncontrolled water and solute removal. However, this approach failed to provide significant results, because ultrafiltration and sodium profiles were predetermined and no adaptations could be made if the designed profile was inadequate. c) The third step in the evolution of dialysis was the understanding that on-line signals from the machine and from the patients were required in order to prepare and carry out the adequate response and variation of treatment parameters. For this reason a series of sensors have been developed including urea and blood volume sensors which are offering the most important signals from the patient. In this way, accurate responses could be made during treatment and from a simple manual feedback, we have today a completely automatic form of biofeedback. The question that now arises is where to find the financial resources to afford the upcoming technology. Another question is whether this new technology should be for everybody in routine dialysis or it should be designed for specific conditions. In other words, are these toys for nice experimental studies and speculations or are they tools to improve dialytic outcomes and morbidity? Probably, technology cannot be stopped in its evolution. What is exceptional today will probably be part of the routine of tomorrow. It seems that we are struggling more with the complex physiology of human body than with mechanical or electronic problems that certainly find their solution before or after. The increasing use of computers and the evolution of the applied software will certainly help in reducing the costs and improving the performances of our newer dialysis devices. PMID- 10688406 TI - Changes in the clinical condition of haemodialysis patients. AB - The characteristics of the dialytic population have substantially changed over the past 30 years, becoming older and with a greater number of coexisting diseases. The considerable evolution in treatment modalities has lead to a significant increase in the efficacy and tolerability of dialysis. However, physicians have to deal with illnesses in long term dialysis survivors that may be a consequence of inadequate renal replacement therapy rather than of the dialysis procedure per se. Cardiovascular diseases are the leading cause of death and, although many of the risk factors are the same as in the general population (i.e. hypertension), some appear to be specific to CRF (i.e. hyperparathyroidism, anaemia). Age is the most important demographic factor associated with increased mortality. The increasing incidence of ESRD diabetic patients, as well as malnutrition, also contribute to higher mortality in RRT. The therapeutic answer to a worsening in clinical condition is adequate medical care (starting in the conservative phase), with particular attention being given to correcting anaemia, hypertension, volume overload and hyperparathyroidism, and preventing malnutrition. Treatment modalities also play a crucial role. Data suggest that adequate dialytic dose (and possibly time) can reduce morbidity and mortality, and on-line sodium and potassium modelling can improve intradialytic cardiovascular stability and reduce arrhythmias. Long-term treatment with synthetic high-flux membranes may confer some beneficial effect on beta2-m amyloidosis-related morbidity and may also reduce mortality. Family and social support greatly affect the quality of life of the patients. However technologically advanced, no procedure can succeed unless it is performed in the context of humanised health care directed towards patient needs. PMID- 10688407 TI - Thirty years of progress in peritoneal dialysis. PMID- 10688408 TI - Systemic hypertension and antihypertensive agents. PMID- 10688409 TI - Renal transplantation, past, present and future. AB - In the absence of immunosuppression, renal transplantation was sporadically and unsuccessfully performed during the first half of this century. Over the past 40 years, immunosuppressive drug regimens have evolved greatly and transformed solid organ transplantation into a routine clinical procedure with a 1-year graft survival between 80% and 90%. The original immunosuppressive scheme was based on the administration of glucocorticoids and azathioprine. However, many patients developed acute rejection which required very high dose of prednisone. As a consequence, a high mortality rate due to opportunistic infections was frequently observed, since this immunosuppressive regimen nonselectively inhibited elements of host resistance such as monocytes, granulocytes, and macrophages. In the early Eighties, the introduction of monoclonal antibodies directed against the CD3 molecule and of cyclosporine, a lymphokine synthesis inhibitor, allowed a more effective control of acute allograft rejection and a more specific target with maintenance immunosuppression. Furtherly, with the knowledge of molecular immunology the better understanding of the cellular and molecular mechanisms that underlie the immunological response to transplanted organs, led to the discovery of new immunosuppressive agents, such as tacrolimus, rapamycin, interleukin-2 monoclonal antibodies, and mycophenolate mofetil. All these drugs showed a more selective mechanism for T- and B-cell alloimmune responses. The results of recent clinical trials based on the combination of these drugs with steroids and cyclosporine reduced the incidence of acute rejection episodes to less than 10% and permitted a steroid-sparing policy in kidney transplantation. Today, the main problem is related to the side-effects of vigorous and prolonged immunosuppression, mainly infections and malignancies. If it were possible to obtain permanent immunological tolerance, immunosuppressive therapy could be minimized. In this respect, the new generation of drugs, FTY 20, antisense oligonucleotides and agents capable of blocking the costimulatory pathway of allorecognition, might have the potential of favoring tolerance in the host against alloantigens. PMID- 10688410 TI - Rapidly progressive glomerulonephritis. AB - The management of rapidly progressive glomerulonephritis has been transformed over the past thirty years. It has become one of the few forms of glomerulonephritis that can be effectively treated, and today overall renal survival is as high as 70%. Effective management of patients with RPGN requires prompt and accurate diagnosis so that patients are appropriately treated, and long term follow up to minimise the risk of relapse in patients with some types of those disease. PMID- 10688411 TI - Diabetic nephropathy--what have we learned in the last three decades? AB - The past three decades have seen enormous conceptual advances in understanding the pathogenesis of diabetic nephropathy. Increasing evidence points to important genetic determination of the renal risk, i.e. the propensity to develop diabetic nephropathy, in type 1 and type 2 diabetic patients. We are also further along the path to understanding the abnormalities of renal hemodynamics that underly these patients' propensity to develop diabetic glomerulosclerosis, i.e. afferent arterial vasodilation and increased glomerular pressure, identified in elegant experimental studies. Another important advance is the recognition that increased urinary albumin excretion is not only an extremely sensitive marker, but also an important player in the pathogenesis of diabetic nephropathy. Finally, the concept of the toxicity of hyperglycemia ("glucotoxicity") has been carried to the molecular level, so that pathomechanisms such as activation of protein kinase C and cellular damage by advanced glycation endproducts (AGE), to name only two, have been elucidated. Diabetic nephropathy has become the leading cause of endstage renal failure (ESRF) in Western countries, particularly in patients with type 2 diabetes. Three treatment modalities are available: (i) hemodialysis,(ii) CAPD and (iii) transplantation, meaning kidney transplantation, combined pancreas and kidney transplantation or - still in a very preliminary stage - islet cell transplantation. The ideal is to have all three modalities available to meet each patient's individual needs. Treatment outcome continues to be considerably worse, however, in diabetic than non-diabetic patients. This highlights the importance of prevention. Progression to ESRF in diabetic nephropathy is preventable, at least to a large extent. PMID- 10688412 TI - Primary glomerulonephritides with nephrotic syndrome. Limitations of therapy in adult patients. AB - Thirty years of clinical studies have shown that a correct therapeutic approach to human glomerulonephritides with nephrotic syndrome requests the evaluation of three important parameters such as renal biopsy, long monitoring of daily proteinuria and renal function. In addition, age and clinical manifestations should be considered. Corticosteroids, alkylating agents (cyclophosphamide, chlorambucil) and purine analogues are currently used in the treatment of primary glomerulonephritis (minimal-change disease (MCD), focal segmental glomerulosclerosis (FSGS), membranous (MGN) and membranoproliferative glomerulonephritis (MPGN)), however results are different. Patients with nephrotic syndrome in MCD when treated with corticosteroids and/or cytotoxic drugs have complete or partial remission in a more than 90% of cases. On the contrary, nephrotic FSGS remits completely or partially only in 50% of treated cases when a more aggressive and prolonged immunosuppressive therapy is carried out. Data from clinical trials in MGN patients are controversial, however it is evident that a greater percentage of patients with stage 1 and stage 2 renal lesions benefit from corticosteroids in association with immunosuppressive drugs. Finally, no encouraging data have been obtained by clinically controlled trials in patients with MPGN. Future perspectives suggest the use of other drugs such as receptor blockade of cytokines and growth factors, administration of cytokine antagonists, intracellular signalling blockade and gene therapy with antisense oligonucleotides. Unfortunately, until specific therapies become available, we have to use unspecific or only symptomatic therapy. PMID- 10688413 TI - Pathogenesis of glomerulonephritis, a perspective from the last 30 years. PMID- 10688414 TI - Pathophysiology of acute renal failure. AB - Acute renal failure (ARF) is a common renal disease affecting up to 5% of all hospitalized patients, with a higher prevalence of 10-30% in patients in critical care units (1-3). Despite advances in the management of critically ill patients and technological advances in renal replacement therapy, the high mortality of patients with ARF has not changed over the last decades and remains above 50% (4 6). Moreover, as a consequence of more advanced medical therapy and more complicated surgical interventions in older and multimorbid patients, the number of patients with ARF is increasing (1, 4, 5). Moreover, ARF itself increases the risk to develop additional complications that can be deleterious. Recently, an independent association between ARF and mortality has been shown in patients following administration of radiocontrast media in an intensive care unit and in patients following cardiac surgery (6, 7). Following radiocontrast media the mortality of patients with ARF was increased five fold and following cardiac surgery sixteen-fold as compared to patients with the same underlying disease without ARF. The pathophysiology of ischemic ARF is reviewed with the emphasis on the following mechanisms: Increased fractional excretion of sodium, Activation of tubuloglomerular feedback, Cytoskeletal disruption, Tubular obstruction, Vascular mechanisms. The following mediators will also be discussed: Calcium, Cysteine proteases, Nitric oxide, Adhesion receptors and integrins. PMID- 10688415 TI - Ischemic nephropathy. AB - Ischemic nephropathy, involving stenotic lesions in the renal arteries, associated with renal insufficiency, is now recognized as a frequent disease. It may be responsible for a significant proportion of end stage renal disease, at least in the Caucasian population. Some non-invasive but reliable techniques such as echo-color-Doppler, gadolinium-enhanced magnetic resonance and spiral CT angiography are now available for diagnosis. Revascularization with either angioplasty, stent or surgery improves renal function in many patients. In the near future systemic and/or local medical therapy will provide better answers for renovascular disease. PMID- 10688417 TI - Cytokine patterns and the effects of a preoperative steroid treatment in the patients with abdominal aortic aneurysms. AB - BACKGROUND: The aim of this study was to investigate the cytokine patterns of patients with abdominal aortic aneurysms and the effects of preoperative steroid administration on surgical stress. METHODS: From January 1996 to August 1996, 20 consecutive patients underwent an elective reconstruction of infrarenal abdominal aortic aneurysms. The patients were randomly divided into two groups consisting of a control group (n=10) and a steroid group (n=10), in whom 1 g of methylprednisolone was intravenously administered two hours before the operation. MEASURES: Interleukin-6 was serially measured and the perioperative parameters including C-reactive protein were compared between both the control and the steroid groups. RESULTS: The interleukin-6 values in the steroid group immediately after declamping, as well as at one and three postoperative days were significantly lower than those in the control group. C-reactive protein values at one postoperative day in the steroid group were also significantly lower than those in the control group. In one patient with a ruptured abdominal aortic aneurysm, the interleukin-6 values were higher than those in the patients undergoing elective surgery throughout the study. CONCLUSIONS: These results thus suggest that preoperative steroid administration using methylprednisolone in patients with abdominal aortic aneurysms appears to reduce surgical stress by decreasing cytokine release. PMID- 10688416 TI - Angiogenesis for the treatment of vascular diseases. AB - Critical ischemia of the limbs or myocardium is frequently accompanied by diffuse distal vascular disease making it unapproachable by conventional revascularization techniques. Pharmacological treatment is available for coronary artery disease but there has been no effective medical therapy for advanced ischemia of the limbs. In the search for alternative treatments for patients with diffuse distal disease, recent developments in vascular biology have directed attention towards use of vascular growth factors. Therapeutic angiogenesis has shown promising results in early clinical studies as shown by improved clinical status and in some cases angiographic studies. We employed an angiogenic strategy that utilizes enhanced vascular endothelial growth factor (VEGF) in a fibrin network, in two patients with critical limb ischemia. Objectively, we were able to demonstrate angiographically the growth of new blood vessels after administration of VEGF and fibrin composite. Fibrin glue provides for the slow release of and prolongs the availability of VEGF, thereby sustaining angiogenesis resulting in improved oxygenation of ischemic tissue. Further investigations are warranted to validate if angiogenesis may increase blood flow in patients with advanced vascular disease. PMID- 10688418 TI - Clinical laboratory monitoring of a synthetic antithrombin agent, argatroban, using high performance liquid chromatography and functional methods. AB - BACKGROUND: Argatroban is a peptidomimetic inhibitor of thrombin which is in clinical trials for thrombotic complications. Clot-based assays measure the cumulative anticoagulant effect of argatroban and its metabolites(s). To monitor the absolute concentrations of argatroban, a specific HPLC method was developed. METHODS: Validation studies included normal volunteers administered with escalating doses of argatroban (ARG 102 Study), patients undergoing coronary interventional procedures (ARG 310), and patients receiving argatroban in conjuction with streptokinase for acute myocardial infarction (ARG 230). Plasma samples were extracted with acetonitrile and reconstituted in a mobile phase. UV detection was made at 333 nm. Calibratrion curves were prepared with known standards of argatroban in normal human plasma. RESULTS: The retention time for argaeroban was 6.0+/-0.5 min and the extraction efficiency was >98% (r2=0.99). In the ARG102 Study, argatroban levels were: 0.84+/-0.23 (day 1), 1.55+/-0.34 (day 2), 2.92+/-0.15 (day 3), and 3.04+/-0.49 (day 4). In the ARG310 trial, the mean argatroban levels were: 0.23+0.09 microg/ml (preinfusion), 5.77+/-0.92 microg/ml (postinfusion/intraprocedure), and 2.23+/-0.29 microg/ml (postprocedure). In the ARG 230 Study, the mean argatroban levels at 2-8 hrs were between 1.5-2.0 microg/ml. Upon completion of the infusion, a time-dependent clearance of argatroban was noted. CONCLUSIONS: Since heparinization, hemodilution and hypofibrinogenemia due to thrombolysis influence the clotting tests, absolute quantitation of argatroban by HPLC in these patients provides a more reliable means of monitoring this anticoagulant and helps in the dosage-optimization of this agent. The current HPLC method is of value in the monitoring of patients who are simultaneously administered with thrombolytic drugs. PMID- 10688419 TI - Single lacunar brain infarction with transient signs versus those with long lasting signs. AB - BACKGROUND: In order to find out the difference between single brain lacunar infarctions with transient signs and those with long-lasting signs, cerebral blood flow studies and blood tests were performed. METHODS: Ten cases of single lacunar infarction with transient signs and 10 of single lacunar infarction with long-lasting signs were studied. Subcortical cystic infarctions with a diameter of less than 1.5 cm were defined as lacunar infarction. Episodes lasting less than 24 hours were classified as transient signs and those lasting 24 hours or more as long-lasting signs. MEASUREMENTS: cerebral blood flows were measured using the stable xenon computed tomography method. The regional cerebral blood flows were measured before and 20 minutes after the intravenous injection of 17 mg/kg acetazolamide. Plasma fibrinopeptide A, platelet factor 4 and beta thromboglobulin concentrations were determined at the Special Reference Laboratories. RESULTS: Blood flows in the cerebral cortex and cerebral white matter contralateral to the lacunar infarction were lower in the group with long lasting signs than in that with transient signs. Cerebrovascular acetazolamide reactivity in the cerebral cortex and white matter contralateral to the lacunar infarction were lower in the group with long-lasting signs than in that with transient signs. Plasma fibrinopeptide A, platelet factor 4 and beta thromboglobulin concentrations were higher in the long-lasting signs group than in that with transient signs. CONCLUSIONS: There may be some differences in pathogenesis between single lacunar infarction with transient signs and those with long-lasting signs. PMID- 10688420 TI - Duplex screening as a method of quality assurance of perioperative thromboembolism prophylaxis. AB - BACKGROUND: Improvements in thrombosis prophylaxis in both the operative and non operative fields aim to reduce further the not inconsiderable residual risk of suffering a deep vein thrombosis or embolism. The goal of the study was to establish the part played in a quality assurance strategy by early diagnosis of a thrombosis and by knowledge of the hospital's internal patient-risk profile in order to counter the unpredictability of thromboembolic complications and make rational decisions about thromboembolism prophylaxis. METHODS: Duplex ultrasonography has been used routinely in trauma surgical patients in Krefeld Hospital since September 1991 as a screening method for diagnosing deep leg and pelvic vein thrombosis prior to mobilisation of the patients. 778 patients were investigated up to March 1997. In the period from September 1991 to September 1994, patients received standardised low-dose prophylaxis with unfractionated heparin (UFH). In October 1994, the prophylaxis regimen was modified by changing the anti-embolism stockings from bidirectional elastic stockings to transverse elastic graduated compression stockings (TED) and by adapting the dosage of the heparin prophylaxis to patient risk, with the use of low molecular weight heparin (LMWH) Certoparin (Mono-Embolex NM) since April 1995. All patients with a deep vein thrombosis were treated immediately with APTT-monitored full heparinisation and immobilisation. RESULTS: In the period from September 1991 to March 1997, an asymptomatic deep vein thrombosis of the lower limbs was diagnosed in 68 cases (8.7%) out of 778 trauma surgical patients by means of routine duplex ultrasound. Using a strategy of duplex screening and immediate anticoagulation/immobilisation, no clinically significant pulmonary emboli occurred in this period. At the same time, the antithrombotic efficacy of the prophylaxis could be improved and assessed objectively by means of duplex screening: with optimal compression stockings and consistent use of risk-adapted UFH prophylaxis, it was possible to reduce the residual thrombosis rate, which was 11.5% (95% CI 7.7-15.2%) with standard UFH prophylaxis, to 8.7 % (95% CI 4.5 12.9%) and ultimately, using the combination of optimal anti-embolism stockings and LMWH prophylaxis, to 6.0% (95% CI 3.0-8.9%) which was significant (p<0.05). The cost-effectiveness analysis resulted in a cost-relation per successfully treated patient of about 1:100 for the diagnosis of a deep vein thrombosis using duplex ultrasound and subsequent heparin treatment compared to the diagnosis and intensive care treatment of a massive pulmonary embolism. CONCLUSIONS: Duplex ultrasound screening for asymptomatic deep vein thrombosis thus proves to be a suitable instrument for internal hospital quality control in thrombosis prophylaxis. Its routine use can be recommended at least in high-risk patients, not only from the medicolegal aspect but also from the purely economic aspect. PMID- 10688421 TI - Inter-observer variation. An alternative method of assessing the role of ultrasonic imaging in clinical decision-making in lower limb arterial disease. AB - BACKGROUND: The aim of the study is to determine the role of duplex scanning in deciding on the final treatment for patients with lower limb arterial disease compared to intra-arterial digital subtraction arteriography (IA DSA). METHODS: Eighty-two patients (55 males, 27 females, mean age 68 years) with lower limb arterial disease had both duplex and arteriography performed. The findings of both modalities were reported by sonographer and radiologist in 164 anonymous reports. Five vascular surgeons (consultant grade) were asked to plan their treatment on the data provided in these reports. For the purposes of statistical analysis, the actual treatment the patient received was used as the reference standard. Both reports and treatment decisions were coded (double blinded). RESULTS: The overall accuracy of duplex scanning to assess arterial disease of the lower limbs showed a sensitivity of 92%, and specificity of 99%. The accuracy of the decisions based on duplex was 84% and kappa with k of 0.62 (95% CI+0.14) compared to the reference standard, while the accuracy of the decision based on arteriography was 85% and kappa 0.63 (95% CI+0.14). The accuracy of decisions based on duplex compared to those based on arteriography (arteriography used as reference standard) was 91% and a kappa of 0.77 (95% CI+0.14). CONCLUSIONS: Duplex scanning is an accurate diagnostic modality, and clinical decisions can be safely based on its findings. PMID- 10688422 TI - Does in situ replacement of a staphylococcal infected vascular graft with a rifampicin impregnated gelatin sealed Dacron graft reduce the incidence of subsequent infection? AB - BACKGROUND: The aim of this study was to treat methicillin-resistant Staphylococcus aureus (MRSA) or S. epidermidis prosthetic vascular graft infections by in situ replacement with a rifampicin bonded Gelsoft graft. METHODS: Interposition grafts were placed in the internal carotid artery of 56 merino sheep and the graft surface directly inoculated with 10(8) colony forming units (CFU) of MRSA (29) or S. epidermidis (27). At three weeks, grafts were harvested and sheep allocated to three groups. In the MRSA infected group, sheep received grafts soaked in 1.2 mg/ml (12), 10 mg/ml (10) and no (7) rifampicin. For S. epidermidis, sheep received grafts soaked in 1.2 mg/ml (10), 10 mg/ml (9) and no (8) rifampicin. There were two deaths, in the MRSA study group, one each from the rifampicin treated groups. The remaining sheep were euthanased and grafts harvested three weeks following regrafting. Grafts at harvests were assessed for perigraft abscess formation, anastomotic disruption and graft thrombosis. Swabs were taken to assess bacterial growth in the perigraft tissues, and external and internal graft surfaces. A 3-5 mm segment of graft was incubated in a broth medium. For S. epidermidis the remainder of the graft was ground and then incubated in a broth medium. RESULTS: For MRSA, no statistical difference between the groups was reached for any of the measured parameters. For S. epidermidis, a significant reduction was reached for total infected specimens in the 10 mg/ml group compared to both control (p<0.001) and 1.2 mg/ml (p<0.005) groups. Graft reinfection was also less likely to occur with S. epidermidis than MRSA. CONCLUSIONS: In conclusion, replacement of S. epidermidis infected vascular grafts with 10 mg/ml rifampicin soaked Gelsoft graft is effective in reducing subsequent S. epidermidis infection. This conclusion cannot be extended to MRSA infected vascular grafts. PMID- 10688423 TI - External support valvuloplasty in the treatment of chronic deep vein incompetence of the legs. AB - BACKGROUND: To evaluate the benefit from external support valvuloplasty in chronic deep vein incompetence of the legs. METHODS: DESIGN: prospective study. PATIENTS: twenty patients 7 primary and 13 secondary (post-thrombotic), with severely symptomatic deep vein incompetence (DVI) of the legs. Preoperative duplex sonography, videophlebography with ambulatory venous pressure measurement. Surgical treatment with external support valvuloplasty with Venocuff (Vaso Products Inc., Sommer-ville, NJ, USA). Postoperative clinical follow-up, duplex and pressure measurements. RESULTS: In primary DVI, symptoms disappeared in all 7 patients, and in secondary DVI in 7 of 13 patients. All reconstructions were competent in primary DVI and in 10 out of 13 in secondary DVI. The follow-up period averaged 19 (6-32) months in primary DVI and 18 (5-31) months in secondary DVI patients. CONCLUSIONS: In severely symptomatic deep vein incompetence of the legs external support valvuloplasty is effective, especially in primary DVI. In secondary DVI the competence of the reconstructions was 78% and the symptoms disappeared in 52%. This means that external valvuloplasty is indicated even in post-thrombotic patients. PMID- 10688424 TI - Intramural hydatid cyst of the abdominal aorta. AB - Intramural vascular hydatid disease is an extremely rare incidental finding. A unique case of vascular wall hydatid cyst simulating aneurysmal dilatation of the abdominal aorta is presented. PMID- 10688425 TI - Inferior mesenteric artery aneurysm. Case report. AB - This paper reports a large inferior mesenteric artery aneurysm discovered incidentally during the work-up in a male patient with a thoracoabdominal aortic aneurysm. Aortography disclosed an aneurysm in the inferior mesenteric artery with a large marginal artery which filled the branches of the coeliac and superior mesenteric arteries retrogradely. The thoracoabdominal aortic aneurysm was reconstructed by a bifurcated aorto-biiliac Dacron graft while inferior mesenteric artery revascularisation was achieved with a PTFE graft, reconstruction being necessary because of its dominant blood supply to all of the viscera. This case highlights the importance of aneurysmal reconstruction when an anomalous arterial supply to the gastrointestinal tract from a dilated inferior mesenteric artery has been demonstrated on a preoperative angiogram. PMID- 10688426 TI - Primary aortoduodenal fistula treated successfully with surgery in a patient with Takayasu's arteritis. AB - Takayasu's arteritis was originally described as a systemic inflammatory arterial disease presenting with occlusive changes. However, it has also been known to cause aneurysm formation. In this report, a patient with Takayasu's arteritis was found to have an aortoduodenal fistula. An emergency operation was carried out with resection of the saccular aneurysm and the fistula. The aorta was reconstructed with a prosthetic graft and the duodenum repaired. A pedicled omental flap was placed between the aorta and the duodenum. The postoperative recovery was uneventful, there was no evidence of persistent bleeding, and the patient was well at the 3-year follow-up. This is the first case in the English language literature of a primary aortoduodenal fistula treated successfully with surgery in a patient with Takayasu's arteritis. PMID- 10688428 TI - Ultraviolet radiation-induced tolerance. PMID- 10688429 TI - Eosinophil activation by eotaxin--eotaxin primes the production of reactive oxygen species from eosinophils. AB - BACKGROUND: The CC chemokine eotaxin has been shown to possess selective chemotactic activity for eosinophils, the major effector cells in allergic inflammation. Reactive oxygen species (ROS) from eosinophils may damage cells or tissue, such as the mucosal epithelium. In this study, we examined the effect of eotaxin on ROS from eosinophils and compared its activity with RANTES and interleukin (IL)-5. Moreover, we examined the signal transduction of eotaxin and the effect of dexamethasone on ROS from eosinophils. METHODS: Eosinophils were isolated by modified CD16-negative selection. ROS in luminol-dependent or lucigenin-dependent chemiluminescence reaction were examined. Calcium ionophore A23187 was added to the mixture of eosinophils with luminol or lucigenin, and then ROS were determined. RESULTS: Eotaxin primed the production of ROS in a dose dependent manner. ROS from untreated eosinophils evoked with calcium ionophore A23187 in luminol-dependent chemiluminescence gave a maximal value of 4957+/-1035 intensity counts (IC) (mean+/-SE, n=7) and an integral value of 15.75+/-3.14 IC (x10(-4)), while eosinophils that were treated with eotaxin gave maximal values of 11 142+/-2300 IC (10 nM) and 29165+/-3718 IC (100 nM) and integral values of 41.07+5.44 IC (x10(-4)) (10 nM) and 152.90+/-22.38 IC (x10(-4))(100 nM). Moreover, eotaxin was less effective as a priming agent with lucigenin-sensitive pathways than luminol-sensitive pathways. Among several kinds of eosinophils activating cytokines and chemokines, the priming effect of eotaxin on RO5 was the most potent. Eotaxin-primed ROS were inhibited by pertussis toxin, which ADP ribolysates G proteins; wortmannin, a phosphatidylinositol-3-kinase inhibitor; and genistein, a tyrosine kinase inhibitor, suggesting the involvement of pertussis toxin-sensitive G proteins, phosphatidylinositol-3-kinase, and tyrosine kinase in the signal transduction of eotaxin. Moreover, dexamethasone inhibited ROS from not only untreated eosinophils but also eosinophils treated with eotaxin. CONCLUSION: Eotaxin may play an important role in the pathogenesis of allergic inflammation through eosinophil activation by priming of eosinophil oxidative metabolism, as well as by involvement in selective eosinophil chemotaxis. PMID- 10688430 TI - Allergy to date fruits: characterization of antigens and allergens of fruits of the date palm (Phoenix dactylifera L.). AB - BACKGROUND: Date-palm (Phoenix dactylifera L.) fruits are eaten daily by most inhabitants of the Middle East and the neighboring countries. Recent reports have indicated that dates are allergenic. This study aimed to investigate the antigenic and allergenic potential of date fruits. METHODS: Date-fruit extracts from eight cultivars were evaluated in skin prick tests (SPT) in an atopic population, used to produce antisera, analyzed by SDS-PAGE, and fractionated by gel-filtration chromatography. Sera from SPT-positive individuals were evaluated by ELISA and RAST, and in anti-igE immunoblot experiments. RESULTS: About 13% of patients were SPT-positive for at least two extracts. SDS-PAGE of whole extracts revealed 15-18 protein bands of 6.5->100 kDa, and Sephacryl S-200 fractions gave distinct peptide bands. RAST and anti-IgE ELISA gave a range of positive results, which could be abrogated when sera were preabsorbed with fruit extracts. IgE immunoblots of different extracts with pooled positive sera revealed different anti-IgE-binding immunoprints. All the positive sera from fruit-allergic and pollen-allergic individuals bound strongly to two anti-IgE reactive bands of 6.5 to 12-14 kDa and 28-33 kDa, respectively, and about 50% of sera bound to a 54-58 kDa band. CONCLUSIONS: These results strongly indicate that 1) date-palm fruit is a potent allergen 2) sera from fruit-allergic as well as pollen-allergic patients recognize common fruit-specific epitopes 3) there is heterogeneity in patient responses to the different extracts. PMID- 10688431 TI - Early increase in urinary leukotriene E4 (LTE4) is dependent on allergen dose inhaled during bronchial challenge in asthmatic subjects. AB - BACKGROUND: Urinary leukotriene E4 (LTE4) excretion is a good marker of the rate of total body production of sulfidopeptide leukotrienes released during allergen challenge. METHODS: Twenty-three subjects with allergic asthma were challenged with inhaled allergen, and the urinary excretion of LTE4 was determined by immunoenzymatic assay (associated with HPLC separation) at various intervals after challenge. RESULTS: Allergen challenge caused an early airway response (EAR) with a drop in FEV1 of 40.3+/-9.9%. This was associated with an increase in urine LTE4 excretion for 0-3 h after allergen inhalation (296+/-225.25 pg/mg creatinine) in comparison with baseline values obtained during the night before challenge (101.02+/-61.97 pg/mg creatinine). Urinary LTE4 excretion was significantly higher in subjects who inhaled a higher dose of allergen during challenge (LTE4 during EAR: 211+/-192 pg/mg creatinine in subjects with inhaled total dose of allergen <0.1 biologic units; 408+/-223 pg/mg creatinine in subjects with inhaled total dose >0.1 biologic units). All subjects showed a late airway response (LAR) to allergen of different severity, from mild (FEV1 fall: 15 20%) to severe (>30%); no correlation was found between the increase in urine LTE4 excreted during LAR (3-7 h after challenge) and the severity of LAR, but only subjects with severe LAR showed a significant increase in LTE4 during LAR in comparison with baseline value. CONCLUSIONS: A release of sulfidopeptide leukotrienes, as evaluated by urinary LTE4 excretion, can be documented during EAR and LAR to allergen in relation to the dose of inhaled allergen, and it can represent a useful index of the events underlying the airway inflammatory responses during allergen challenge. PMID- 10688432 TI - Overexpression of CD44 on alveolar eosinophils with high concentrations of soluble CD44 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia. AB - BACKGROUND: High levels of interleukin-5 (IL-5) are found in the alveolar space of patients with eosinophilic pneumonia (EP). IL-5 promotes the growth and differentiation of eosinophils, as well as activating these cells. IL-5 also induces the expression of CD44 on eosinophils in vitro. To evaluate the contribution of CD44 to the pathogenesis of EP, we examined the expression of CD44 on eosinophils in bronchoalveolar lavage fluid (BALF) and measured the concentration of soluble CD44 (sCD44) in BALF from patients with EP. METHODS: The concentrations of IL-5, sCD44, and hyaluronic acid (HA) were measured in BALF. The expression levels of CD44 on eosinophils in BALF and peripheral blood in patients with EP were compared. RESULTS: The expression of CD44 on alveolar eosinophils and the concentration of sCD44 were increased in BALF of patients with EP. There was a significant correlation between IL-5 and sCD44 in BALF. A high concentration of HA was observed in BALF of EP patients. CONCLUSIONS: Our results suggest that the high expression of CD44 on eosinophils probably results from upregulation by IL-5 and could be important in the pathogenesis of EP. PMID- 10688427 TI - Biologic functions of the IFN-gamma receptors. AB - Interferon-gamma (IFN-gamma) is a cytokine that plays an important role in inducing and modulating an array of immune responses. Cellular responses to IFN gamma are mediated by its heterodimeric cell-surface receptor (IFN-gammaR), which activates downstream signal transduction cascades, ultimately leading to the regulation of gene expression. In order to study the role of IFN-gamma in a number of immune responses and pathways, researchers have generated mice with altered patterns of IFN-gammaR gene expression. These studies, together with analyses of naturally occurring mutations of the IFN-gammaR in man, have been instrumental in elucidating the diverse functions of IFN-gamma, and are the subject of this review. PMID- 10688433 TI - Regulation of ICAM-3 and other adhesion molecule expressions on eosinophils in vitro. Effects of dexamethasone. AB - BACKGROUND: ICAM-3 has been recently identified as the third leukocyte-function associated-1 (LFA-1) ligand. ICAM-3 is expressed in eosinophils, but its regulation has not been studied. The objective of this study was to investigate the differential expression of ICAM-3 and other adhesion molecules (AM) on the surface of eosinophils. We also evaluated the effects of dexamethasone on AM expression. METHODS: Normodense eosinophils were isolated from peripheral blood and incubated with calcium ionophore A23187 (calcimycin) with and without dexamethasone. Expression of AM was assessed by flow cytometry and expressed as fluorescence mean intensity (FMI). RESULTS: Peripheral blood eosinophils constitutively expressed low levels of ICAM-1 and ICAM-2 (<10 FMI), moderate levels (10-50 FMI) of CD29 and L-selectin, and high levels (>50 FMI) of ICAM-3, LFA-1, and Mac-1. Calcium ionophore (1 microM) significantly increased Mac-1 and ICAM-1 expression at 6 and 24 h. L-selectin expression decreased at 6 and 24 h, but ICAM-2, ICAM-3, LFA-1, and CD29 expression did not show any significant change after calcium ionophore stimulation. Dexamethasone decreased ICAM-3 and increased L-selectin basal expression, and it caused a dose-related inhibition of calcium ionophore-induced ICAM-1 expression. CONCLUSIONS: These findings suggest that some AM, such as ICAM-1, Mac-1, and L-selectin, may be involved in adhesion during eosinophil activation and that glucocorticoids may prevent airway inflammation by regulating the expression of AM in eosinophils. The role of ICAM 3, a leukocyte AM highly expressed in resting eosinophils, remains to be clarified. PMID- 10688434 TI - Different serum soluble Fas levels in patients with allergic rhinitis and bronchial asthma. AB - BACKGROUND: The pathogeneses of allergic rhinitis and bronchial asthma are believed to be closely mutually related because of the similar dynamics of allergy-inducing cells and molecules and clinical overlap. In this study, we compared these diseases in the dynamics of cell apoptosis-regulating molecules. METHODS: Allergic rhinitis patients (n=36), bronchial asthma patients (n=22), and healthy subjects (n=32) were subjected to measurement of serum (soluble Fas) (sFas) levels during the stable and attack disease phases by a sandwich enzyme linked immunosorbent assay. RESULTS: Serum sFas levels in patients with allergic rhinitis during the attack phase were significantly lower (P<0.0001) than those in healthy individuals. There were no differences between them during the attack and stable disease phases. In contrast, serum sFas levels in patients with bronchial asthma during the attack phase were higher (P<0.0005) than those in healthy individuals. Interestingly, the levels during the attack phase were lower (P<0.002) than those during the stable phase. CONCLUSIONS: Our results suggest a different pathogenesis for allergic rhinitis and bronchial asthma at the cell apoptosis-linked step. PMID- 10688435 TI - Integrin expression on neutrophils and mononuclear cells in blood and induced sputum in stable asthma. AB - BACKGROUND: We speculated that the expression of integrins in the airway lumen of asthmatic subjects might be altered compared with normal subjects during cell recruitment from circulation. METHODS: To test this hypothesis, we investigated the expression of integrin alpha-chains (CD11a, CD11b, and CD11c) in hypertonic saline-induced sputum and peripheral blood leukocytes in mild to moderate stable asthmatic and control subjects. Immunoreactivity for integrin alpha-chains was assessed by immunocytology on cytospin preparations of sputum and blood. RESULTS: In comparison of the percentages of CD11a+, CD11b+ and CD11c+ mononuclear cells in sputum with their blood counterparts, no significant differences were observed in control subjects, whereas CD11a and CD11b in asthmatic subjects were less expressed on sputum cells. In both control and asthmatic subjects, sputum neutrophils tended to decrease their expression of integrin alpha-chains compared with circulating neutrophils. CONCLUSIONS: We showed that the sputum of asthmatics, unlike normal subjects, is characterized by decreased expression of integrins on mononuclear cells compared with their blood counterparts. The results suggest that downregulation of integrins occurs in asthmatic airways after cell recruitment from circulation. PMID- 10688436 TI - Aeropollinic sampling at three different heights by personal volumetric collector (Partrap FA 52). AB - BACKGROUND: Aeropollinic sampling is usually performed by volumetric pollen traps located on the top of buildings at a height of 15-20 m. The present study aimed to determine whether pollen concentration is similar, at the same time, at different heights. METHODS: Pollen concentrations were measured at the same time for 3 days each month (March-October 1997), with three Partrap FA 52 devices respectively located at 1.5, 5, and 15 m above the ground. A Burkard sampler was used as control at the 15-m level. RESULTS: No significant difference in either the total count or the single pollen counts was observed between the Partrap and Burkard samplers at 15 m. The total pollen count did not differ significantly between the Partrap at 1.5 m and 15 m (130+/-19 and 123+/-18, respectively), whereas the sampler at 5 m collected an amount of pollen (84+/-14) significantly lower than at 1.5 and 15 m (P<0.0001 and P<0.001, respectively). The amount of Urticaceae pollen was significantly higher at 1.5 m (97+/-17) than at 15m (80+/ 12) and 5m (58+/-11); P<0.02 and P<0.001, respectively. Regarding grass pollen, the collector at 5 m captured significantly less pollen (4.5+/-0.8) than at 1.5m (9.5+/-1.3) and 15m (7.2+/-1.3) (P=0.002 and P=0.02, respectively). No significant difference was observed between the data obtained from samplers at 1.5 and 15 m. In addition, the Oleaceae, Cupressaceae, and Corylaceae pollen counts were significantly higher when collected at 15 than at 5 and 1.5 m. CONCLUSIONS: These data show that differences exist in pollen sampling performed at different heights. PMID- 10688437 TI - Allergic contact gingivostomatitis from a temporary crown made of methacrylates and epoxy diacrylates. AB - Occupational allergic contact dermatitis caused by (meth)acrylates is common in dental personnel, whereas dental acrylic fillings and crowns have rarely been reported to cause problems in dental patients. Here we report on a 48-year-old woman who developed gingivitis, stomatitis, and perioral dermatitis after a temporary crown made of restorative, two-component material had been inserted. The manufacturer stated that the temporary crown base paste and catalyst contained three (meth)acrylates, namely, a proacrylate, which is a modification of 2,2-bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (BIS-GMA); a tricyclate, which is a saturated, aliphatic, tricyclic methacrylate; and urethane methacrylate. The manufacturer refused to give more exact information on the (meth)acrylates. Patch testing revealed that the patient was highly allergic to BIS-GMA, other epoxy diacrylates, and (meth)acrylates, as well as to the base paste and catalyst of the temporary crown. Accordingly, it was concluded that the allergic reaction was caused by BIS-GMA, or a cross-reacting (meth)acrylate, or other (meth)acrylates in the temporary crown. PMID- 10688438 TI - A case of allergy to cow's milk hydrolysate. AB - We here report a girl, now 3 years old, who has suffered from severe food allergy since her first year of life. She was strongly allergic to cow's milk, and had high levels of IgE antibody (AB) to casein (210 kU/I), beta-lactoglobulin (43 kU/I), and alpha-lactalbumin (23 kU/l) at 12 months of age. In addition, at the same age, she showed positive (2-4+) skin prick reactions to both unboiled and boiled formulas (Profylac, Nutramigen, and Neocate), besides being positive in RAST to Nutramigen (0.6 kU/l). During the first 3 years, IgE Ab levels against casein and Nutramigen increased to 310 and 1.6 kU/l, respectively. Furthermore, at 3 years of age, she had positive RAST to 14 of 15 tested food allergens, being negative only to codfish. Assessment of eosinophil-related markers revealed high total eosinophil count, increased eosinophil activity, and a low ratio of interferon (IFN)-gamma:IL-5, indicating enhanced IL-5 production. The food allergy was correlated to poor weight gain and increasing problems with atopic allergy in the airways. PMID- 10688439 TI - Circulating soluble L-selectin in atopic dermatitis. PMID- 10688440 TI - Anticonvulsant hypersensitivity syndrome due to carbamazepine. PMID- 10688441 TI - Anaphylaxis due to Monascus purpureus-fermented rice (red yeast rice). PMID- 10688442 TI - Cross-reacting allergens in natural rubber latex and jelutong. PMID- 10688443 TI - Sensitization to aeroallergens in Ankara, Turkey. PMID- 10688444 TI - Occupational asthma to perfume. PMID- 10688446 TI - Virus problems. PMID- 10688445 TI - Unusual egg allergy in an adult. PMID- 10688447 TI - Sparkling clean in Idaho. PMID- 10688448 TI - Heating and pain sensation produced in human skin by millimeter waves: comparison to a simple thermal model. AB - Cutaneous thresholds for thermal pain were measured in 10 human subjects during 3 s exposures at 94 GHz continuous wave microwave energy at intensities up to approximately 1.8 W cm(-2). During each exposure, the temperature increase at the skin's surface was measured by infrared thermography. The mean (+/- s.e.m.) baseline temperature of the skin was 34.0+/-0.2 degrees C. The threshold for pricking pain was 43.9+/-0.7 degrees C, which corresponded to an increase in surface temperature of approximately 9.9 degrees C (from 34.0 degrees C to 43.9 degrees C). The measured increases in surface temperature were in good agreement with a simple thermal model that accounted for heat conduction and for the penetration depth of the microwave energy into tissue. Taken together, these results support the use of the model for predicting thresholds of thermal pain at other millimeter wave (length) frequencies. PMID- 10688449 TI - Models for retrospective quantification of indoor radon exposure in case-control studies. AB - In epidemiologic studies on lung cancer risk due to indoor radon the quantification of individual radon exposure over a long time period is one of the main issues. Therefore, radon measurements in one or more dwellings, which in total have been inhabited by the participants for a sufficient time-period, are necessary as well as consideration of changes of building characteristics and ventilation habits, which influence radon concentration. Given data on 1-y alpha track measurements and personal information from 6,000 participants of case control studies in West and East Germany, an improved method is developed to assess individual radon exposure histories. Times spent in different rooms of the dwelling, which are known from a personal questionnaire, are taken into account. The time spent outside the house (average fraction 45%) varies substantially among the participants. Therefore, assuming a substantially lower radon exposure outside the dwelling, the residence time constitutes an important aspect of total radon exposure. By means of an analysis of variance, important determinants of indoor radon are identified, namely constant conditions such as type of house (one family house or multiple dwelling), type of construction (half-timbered, massive construction, lightweight construction), year of construction, floor and type of basement, and changeable conditions such as heating system, window insulation, and airing habits. A correction of measurements in former dwellings by factors derived from the analysis is applied if current living conditions differ from those of the participants at the time when they were living in the particular dwellings. In rare cases the adjustment for changes leads to a correction of the measurements with a factor of about 1.4, but a reduction of 5% on average only. Exposure assessment can be improved by considering time at home and changes of building and ventilation conditions that affect radon concentration. The major concern that changes in ventilation habits and building conditions lead to substantial errors in exposure (and therefore risk) assessment cannot be confirmed in the data analyzed. PMID- 10688450 TI - Soil-to-plant transfer factors for radiocesium and radiostrontium in agricultural systems. AB - A database of soil-to-plant transfer factors for radiocesium and radiostrontium has been compiled for arable crops from published and unpublished sources. The database is more extensive than previous compilations of data published by the International Union of Radioecologists, containing new information for Scandinavia and Greece in particular. It also contains ancillary data on important soil characteristics. The database is sub-divided into 28 soil-crop combinations, covering four soil types and seven crop groups. Statistical analyses showed that transfer factors for radiocesium could not generally be predicted as a function of climatic region, type of experiment, age of contamination, or soil characteristics. However, significant relationships accounting for more than 30% of the variability in transfer factor were identified between transfer factors for radiostrontium and soil pH/organic matter status for a few soil-crop combinations. Best estimate transfer factors for radiocesium and radiostrontium were calculated for 28 soil-crop combinations, based on their geometric means: only the edible parts were considered. To predict the likely value of future individual transfer factors, 95% confidence intervals were also derived. A comparison of best estimate transfer factors derived in this study with recommended values published by the International Union of Radioecologists in 1989 and 1992 was made for comparable soil-crop groupings. Whilst there were no significant differences between the best estimate values derived in this study and the 1992 data, radiological assessments that still use 1989 data may be unnecessarily cautious. PMID- 10688451 TI - A stochastic model of radiation-induced bone marrow damage. AB - A stochastic model, based on consensus principles from radiation biology, is used to estimate bone-marrow stem cell pool survival (CFU-S and stroma cells) after irradiation. The dose response model consists of three coupled first order linear differential equations which quantitatively describe time dependent cellular damage, repair, and killing of red bone marrow cells. This system of differential equations is solved analytically through the use of a matrix approach for continuous and fractionated irradiations. The analytic solutions are confirmed through the dynamical solution of the model equations using SIMULINK. Rate coefficients describing the cellular processes of radiation damage and repair, extrapolated to humans from animal data sets and adjusted for neutron-gamma mixed fields, are employed in a SIMULINK analysis of criticality accidents. The results show that, for the time structures which may occur in criticality accidents, cell survival is established mainly by the average dose and dose rate. PMID- 10688452 TI - Monte Carlo calculation of dose rate conversion factors for external exposure to photon emitters in soil. AB - The dose rate conversion factors D(CF) (absorbed dose rate in air per unit activity per unit of soil mass, nGy h(-1) per Bq kg(-1)) are calculated 1 m above ground for photon emitters of natural radionuclides uniformly distributed in the soil. Three Monte Carlo codes are used: 1) The MCNP code of Los Alamos; 2) The GEANT code of CERN; and 3) a Monte Carlo code developed in the Nuclear Technology Laboratory of the Aristotle University of Thessaloniki. The accuracy of the Monte Carlo results is tested by the comparison of the unscattered flux obtained by the three Monte Carlo codes with an independent straightforward calculation. All codes and particularly the MCNP calculate accurately the absorbed dose rate in air due to the unscattered radiation. For the total radiation (unscattered plus scattered) the D(CF) values calculated from the three codes are in very good agreement between them. The comparison between these results and the results deduced previously by other authors indicates a good agreement (less than 15% of difference) for photon energies above 1,500 keV. Antithetically, the agreement is not as good (difference of 20-30%) for the low energy photons. PMID- 10688453 TI - Verification and modification of the ICRP-67 model for plutonium dose calculation. AB - On the basis of the available data and empirical expressions for the plutonium excretion after injection, an age-related compartmental model has been developed. It provides a better agreement with measured urinary excretion data than the current ICRP 67 model. Moreover, the revised model avoids unphysiological assumptions such as the transfer of activity from soft tissue to urinary bladder, that were part of the ICRP model. The new predictions of the activity in feces and in blood after an injection are closer to the available data than the ICRP 67 estimations and there is also a good agreement with the partitioning of plutonium between skeleton and liver obtained from different autopsy studies. Furthermore, the urinary excretion estimated by the improved model has been checked using some data from occupationally exposed individuals. As the plutonium uptake in these workers occurred by inhalation, the improved model and the ICRP 67 model were compared by connecting them to the ICRP 66 respiratory tract model. The improved model consistently yields a better agreement with the measured excretion and higher estimations of intake than the ICRP 67 model. PMID- 10688454 TI - Gamma radiography with 75Se sources: consequences of a violent fire. AB - Due to the volatile nature of selenium and selenium dioxide, an important atmospheric dispersion of 75Se can occur in case of a violent fire (>800 degrees C). This type of accident is possible when transporting or using a highly active 75Se source for gamma radiography. The consequences of this dispersion are analyzed. From the point of view of radiological protection, it can be concluded that in radiography the unprotected 75Se sealed sources have to resist for 1 h to a temperature of 1,200 degrees C. If this is not the case, x-ray equipment and 192Ir sources have to be preferred above 75Se sources. PMID- 10688455 TI - Comparison of two dose-area-product ionization chambers with different conductive surface coating for over-table and under-table tube configurations. AB - A custom-built graphite-coated transmission ionization chamber is compared to the VacuDAP 2001 (VacuTec, Dresden, Germany), which has transparent conductive electrodes. A study was made of the dependence of response on x-ray tube potential for both types of chamber under identical conditions of exposure using over-table and under-table x-ray tubes. Since the calibration factor is the dose area product of the radiation incident on the patient per chamber reading, it depends on the intrinsic response of the chamber as well as the effect of material in the beam between the x-ray tube and patient. Differences of about 20% were measured between the intrinsic and the over-table calibration factors and between the over-table and the under-table calibration factors for both chambers. The VacuDAP display is specifically calibrated for the over-table condition and would overstate the actual DAP in the under-table case. The intrinsic response of the graphite chamber is nearly independent of tube potential. Although the variation of response with tube potential of the graphite chamber is increased when it is used as an over-table and an under-table patient monitor, it shows less overall variation of response than the VacuDAP. The average deviation of each calibration factor from the mean is less than 5% over the range of 40 to 140 kVp for both chambers. PMID- 10688456 TI - Applications of a quadratic variance model for counting data. AB - A quadratic variance model expressed as a function of sample mean is used to describe counting variance for a mechanical system that exhibits extra-Poisson variance. The nonlinear term describes the extra-Poisson variance, and the linear terms describe the intrinsic and propagated Poisson variance. The quadratic variance model also is applied to repetitive bioassay data, where the nonlinear term describes the well-known phenomenon of biological variance, which is a special case of extra-Poisson variance. The model was found to be suitable for the bioassay data as well. Detection limits for extra-Poisson variance are discussed, as well as the estimation of net signal detection limits, intake, and committed effective dose equivalent using the quadratic variance model. PMID- 10688457 TI - Non-invasive measurement of water content in a lung phantom by neutrons. AB - A novel, non-invasive technique for measuring water content in lungs is described. This technique could find important medical applications such as real time monitoring of lung edema development in patients affected by cardiac insufficiency undergoing physical effort. The technique is based on the moderation (speed decrease) of fast neutrons crossing the lungs, due mostly to the water contained in lungs. In the application of this diagnostic method, the necessary dose given to the patient is lower than that associated with lung imaging techniques that do not use an image intensifier. The results so far obtained are encouraging, and the method appears to be an improvement over those so far reported in literature as it is able to detect a lung water increase equivalent to 25% of a full edema with a 20% standard error. Research is being performed in order to refine the technique, investigate geometry effects and make the method suitable to the medical field. PMID- 10688458 TI - A method for determining leakage of 133Xe gas from septum-sealed glass vials. AB - We have developed a method for determining the leakage of 133Xe gas from septum sealed glass vials that are supplied for medical examinations. Twenty vials each originally containing 370 MBq of 133Xe and 20 vials each originally containing 740 MBq 133Xe were measured daily for 26 d. Retention of 133Xe within the vial was modeled as a first order process with a constant rate coefficient, lambdaT. The value of lambdaT was estimated for each vial using a regression analysis. The leakage rate, lambdaL, was then determined assuming that lambdaT = lambdaL + lambda(r) where lambda(r) represents the physical decay of 133Xe. Monte Carlo simulations were performed using uncertainties in the estimates of each vial to obtain the mean and tails of the distribution for the average leakage rate, lambdaL. The average leakage rate for the complete sample of vials was 0.00007 d( 1) with an upper, one-sided, 95% confidence limit of 0.0011 d(-1). Uncertainties in the published values of lambda(r) for 133Xe made a significant contribution to the uncertainties of the leakage rate for this sample of vials. The methods described can be applied to other situations where leakage of radioactive materials may be of concern. PMID- 10688459 TI - Radiation safety program outcomes as indicated by regulatory compliance activities from 1988 to 1997 in Texas. AB - Occupational radiation protection programs rarely encounter readily observable workplace injuries or illnesses, so upper management must rely on different indicators of relative performance. In many cases, the number of violations, complaints, and reported incidents is used. As with reported workplace injury and illness data, violation, complaint, and incident data provide only a crude indication of a radiation protection program's overall effectiveness. Even so, it is important to recognize that tangible program outcome measures such as these often dictate management decisions. Hence, safety professionals should have access to baseline violation, complaint, and incident trend data so that more informed preventive strategies can be put into place where possible. To assess the outcomes of radiation protection programs in Texas, data from regulatory compliance activities for a 10-y period, inclusive of calendar years 1988 to 1997, were assembled, summarized, and independently verified. For licensees of radioactive material, the ten most frequently cited violations represented 64% of the total issued during the study period. For registrants of radiation producing devices, the ten most frequently cited violations accounted for 73% of the total. A particular emphasis on proper recordkeeping is evident, and should be noted by any internal radiation protection quality assurance programs. Regardless of the permit type, the vast majority of violations issued were classified as low severity. Licensees were found to be involved in approximately 73% of the incidents recorded, with overexposures and badge overexposures representing the largest identifiable types. Registrants were found to be involved in approximately 57% of the complaints recorded, with the largest identifiable issue being concerns about health care provider qualifications or performance. Although this study was limited to a single state, the results can be of benefit to both quality assurance programs and professional health physics training courses by objectively identifying the areas commonly found to be deficient. PMID- 10688460 TI - Technology-based distance education and the absence of physical presence. PMID- 10688461 TI - Kansas City Colleagues in Caring: giving new meaning to networking. PMID- 10688462 TI - LPN to RN articulation: a collaborative solution. PMID- 10688463 TI - A framework for assessing outcomes and practices in Web-based courses in nursing. AB - This article presents a framework to assess the dynamic interaction of technology used to offer Web-based courses, the teaching-learning practices in these courses, and the outcomes enabled by the technology. Concepts of the model include outcomes, educational practices, faculty support, learner support, and use of technology. Variables are identified for each of the concepts. PMID- 10688464 TI - Insights learned from teaching pathophysiology on the World Wide Web. AB - This article describes a one-credit, graduate level pathophysiology module taught using the World Wide Web. Student outcomes are compared to those of students who took the same module in a traditional classroom setting. Although the majority of the graduate students were not Web literate, they became more comfortable with this instructional medium over time. A comparison of the Web-based instruction with the traditional format, both directed by the same instructor, showed no significant differences in student performance on a multiple choice examination. PMID- 10688465 TI - Teaching using interactive video: creating connections. AB - This article addresses the two essential elements of distance learning: the technology and the pedagogy. Both areas are discussed through the four components -information, support, resources, and relationships--of a work effectiveness model. Drawing from recent literature and their experience, the authors offer strategies for making interactive video technology "invisible" while engaging students at a distance. Students experience connections when faculty know how to manage the equipment, plan ahead, and consciously construct strategies for creating relationships across the miles. PMID- 10688466 TI - Graduate education in nursing leadership through distance technologies: the Canada-Norway Nursing Connection. AB - The Canada-Norway Nursing Connection was a collaborative project designed to provide an international educational experience for graduate students in nursing via distance technology. Computer-conferencing and video-teleconferencing were used to address nursing leadership content through case studies. The same technologies were employed to develop the project. The processes of planning and implementing the international linkage are described. Agreement about goals, content, context for online discussion, delivery methods, academic expectations, language support, and logistics was essential. The media proved to be effective for students to gain understandings about nursing leadership, health care, and the forces influencing the nursing profession globally. Insights from the project provided a basis for the development of a model for interactive, international graduate education that will be of value to educators dedicated to helping students gain a global understanding of nursing and health care issues. PMID- 10688467 TI - Lessons learned: using asynchronous computer-mediated conferencing to facilitate group discussion. PMID- 10688468 TI - A pilot study to investigate the impact of interactional television on student evaluation of faculty effectiveness. AB - While the dominant theme of distance education research has been the learner, learner achievement is not necessarily the only important consideration in assessing the effectiveness of ITV programs. With nearly half of the nursing programs recently surveyed by Reinert and Fryback (1997) offering or planning to offer distance learning in the near future, researchers and developers must seek to understand the impact of ITV on the teacher. These programs require faculty who are committed and prepared to teach outside of the traditional classroom. Selection of faculty requires attention to appropriate qualifications and knowledge of strategies to bridge teacher-learner separation created by distance education. Faculty, peers, and administrators should consider the teaching environment when interpreting data designed to evaluate teacher effectiveness of ITV teaching. PMID- 10688469 TI - Evaluating electronic information strategies in a master of science in nursing and master in health services administration interdisciplinary learning experience. PMID- 10688470 TI - Head restraints--the neglected countermeasure. AB - In a rear-end crash, if an occupant's head is unsupported it lags behind as the torso is accelerated forward. This causes the neck to change shape, first taking an s-shape and then bending backward in a 'whiplash' motion. This sudden differential movement of the head and torso can cause 'whiplash' injuries to the neck. This paper reviews methods to minimize the differential head/torso movement and reduce the resulting injuries, focusing on the necessary first step for prevention, which is a head restraint that is behind and close to the back of an occupant's head during the crash. The history of head restraints since the 1950s is reviewed, with particular attention to advanced restraint designs that are proving effective in reducing whiplash injury risk in dynamic tests using a new crash test dummy neck and a new neck injury criterion. PMID- 10688471 TI - Is a lifetime history of neck injury in a traffic collision associated with prevalent neck pain, headache and depressive symptomatology? AB - The objective of this study is to determine whether independent associations exist between a history of neck injury related to a motor vehicle collision and: (1) graded neck pain in the past 6 months; (2) headaches in the past 6 months and; (3) depressive symptomatology during the past week. We used data from the Saskatchewan Health and Back Pain Survey, a population-based cross-sectional survey mailed to a stratified random sample of 2184 Saskatchewan adults aged 20 69 years. Fifty-five percent of the eligible population participated. The exposure was collected by asking subjects whether they had ever injured their neck in a motor vehicle collision. The outcomes: 6-month prevalence of graded neck pain, 6-month prevalence headache and depressive symptomatology during the past week were measured with valid and reliable questionnaires. Sixteen percent of the study sample reported a lifetime history of neck injury in a traffic collision. The association between neck injury and the outcomes was determined from polytomous and binary multivariate logistic regression with adjustment for age, gender and other covariates. A history of neck injury was positively associated with low intensity/low disability neck pain (OR = 2.81; 95% CI 1.81 4.37), positively associated with high intensity/low disability neck pain (OR = 4.46; 95% CI 2.49-4.99) and with disabling neck pain (OR = 3.30; 95% CI 1.48 7.39). Similarly, we found a positive association between a history of neck injury in a motor vehicle collision and headaches that moderately/severely impact on one's health (OR = 2.09; 95% CI 1.27-3.44). No association was found between neck injury and depressive symptomatology (OR = 0.84; 95% CI 0.50-1.40). Our cross-sectional analysis suggests that neck pain and severe headaches are more prevalent in individuals with a history of neck injury from a car collision. However, the results should not be used to infer a causal relationship between whiplash and chronic neck pain and headaches. PMID- 10688472 TI - Biomechanical assessment of soft tissue cervical spine disorders and expert opinion in low speed collisions. AB - The multidisciplinary research of injury mechanisms and injury prevention requires the assessment of the technical and biomechanical circumstances of a collision; moreover, the causality assessment in the individual cases is facilitated by taking these aspects into account. In fact, only specially trained engineers and biomechanical experts are in a position to evaluate these relevant basic facts. In many crucial court cases, important technical factors such as collision angle, structural stiffness, extent of intrusion and the vehicle's velocity change are often ignored. The purely medical causality assessment is often based only on a coincidence of time of the 'accident' and the onset of the disorders. Unfortunately, statements about the 'accident speed' or the nebulous 'accident energy' are often made by clinicians with neither a proper collision documentation nor the necessary biomechanical and technical background. In order to overcome shortcomings of injury causality assessment as well as the terminology associated with soft tissue cervical spine injuries, a subdivision of the term 'accident severity' into four classes is proposed. Consequently, an 'accident severity assessment' can only be performed by a collaboration of four corresponding classes of experts, i.e. the engineer (dynamic loading of the vehicle), the biomechanical expert (biomechanical loading of the occupant), the physician (clinically diagnosable injuries), and eventually the psychiatrist (subjective sequelae individually experienced by the victim). PMID- 10688473 TI - Neck injuries in car collisions--a review covering a possible injury mechanism and the development of a new rear-impact dummy. AB - A review of a few Swedish research projects on soft tissue neck injuries in car collisions is presented together with some new results. Efforts to determine neck injury mechanisms was based on a hypothesis stating that injuries to the nerve root region in the cervical spine are a result of transient pressure gradients in the spinal canal during rapid neck bending. In experimental neck trauma research on animals, pressure gradients were observed and indications of nerve cell membrane dysfunction were found in the cervical spinal ganglia. The experiments covered neck extension, flexion and lateral bending. A theoretical model in which fluid flow was predicted to cause the transient pressure gradients was developed and a neck injury criterion based on Navier-Stokes Equations was applied on the flow model. The theory behind the Neck Injury Criterion indicates that the neck injury occurs early on in the rearward motion of the head relative to the torso in a rear-end collision. Thus the relative horizontal acceleration and velocity between the head and the torso should be restricted during the early head-neck motion to avoid neck injury. A Bio-fidelic Rear Impact Dummy (BioRID) was developed in several steps and validated against volunteer test results. The new dummy was partly based on the Hybrid III dummy. It had a new articulated spine with curvature and range of motion resembling that of a human being. A new crash dummy and a neck injury criterion will be very important components in a future rear-impact crash test procedure. PMID- 10688474 TI - Whiplash injury--are current head restraints doing their job? AB - It is generally accepted that the incidence of whiplash associated disorders is increasing in all industrialised countries, despite the almost universal fitment of head restraints in at least the front seats of cars. This is usually attributed to the fact that few people can be observed to follow the standard recommendations as regards head restraint positioning, that is, level with the head vertically and as close to the head as possible horizontally. This study set out to determine whether any other factors, in addition to head restraint adjustment, could be found which would influence the severity of whiplash injury. This was done by linking medical assessment of real-world accident victims with engineering assessment of the accident vehicles. A random sample of road accident victims suffering from whiplash associated disorder was studied. The vehicles they had been travelling in were examined to assess impact severity and, where possible, measurements were made of seat and head restraint adjustment with the subject sitting in the vehicle. All subjects were interviewed to assess the disability resulting from their injuries, and their progress was followed for 12 months. The results were subjected to statistical analysis to try to determine relationships between severity of injury (as measured by resultant disability) and a number of occupant- and vehicle-related factors. A significant proportion of the sample had suffered lumbar strain injury in addition to whiplash, and these were excluded from the present analysis. Frontal impact victims suffered symptoms indistinguishable from those of rear impact victims. The beneficial effects of good head restraint adjustment could not be clearly demonstrated, and some trends, especially in rear impacts, where the benefits of a well-adjusted restraint should have been very clear, indicated that larger distances from head to restraint were associated with lower disability. The paper discusses these counter-intuitive results and their implications. PMID- 10688475 TI - How crash severity in rear impacts influences short- and long-term consequences to the neck. AB - The main public-health problem concerning WAD are injuries leading to long-term consequences. Yet epidemiological studies mostly concentrate on data based on the injury outcome occurring shortly after the crash. The purpose of this article is to study the influence of crash severity in rear impacts leading to short and long-term consequences to the neck (WAD 1-3), lasting less than or more than 1 year. The influence of change of velocity as well as the car acceleration were investigated by using data from crash pulse recorders (CPR) installed in vehicles, involved in rear impacts. The influence of the car acceleration were also investigated by studying the frequency of occurrence of a tow-bar (hinge) on the struck car. Apart from real-life data, full-scale car-to-car crashes were performed to evaluate the influence of a tow-bar on the struck car. The crash tests showed that a tow-bar may significantly affect the acceleration of the car as well as that of the occupant. According to real-life crashes, a tow-bar on the struck car increased the risk of long-term consequences by 22% but did not affect the risk of short-term consequences. Out of the 28 crash recorder-equipped struck cars involving 38 occupants, 15 sustained no injury where the peak acceleration was 6g or less, 20 sustained short-term consequences where the peak acceleration was 10g or less. Three occupants from two different crashes sustained long-term consequences. The two crashes which resulted in long-term disabling neck injuries had the highest peak acceleration (15 and 13 x g), but not the highest change of velocity. PMID- 10688476 TI - Neck injuries in frontal impacts: influence of crash pulse characteristics on injury risk. AB - AIS1 neck injuries are the most frequent disabling injuries among car occupants in road traffic accidents. Although neck injury is mostly regarded as resulting from rear end collisions, almost one third of all neck injuries occur in frontal impacts. The injury mechanisms in both rear-end and frontal impacts are still not known, although different hypotheses exist. Since 1992, approx. 100,000 vehicles on the Swedish market have been equipped with crash recorders to measuring frontal impacts. This paper analyses the influence of different characteristics derived from the acceleration time history on the risk of short- and long-term disability to the neck in frontal impacts. The study includes injury outcomes from 187 restrained front seat occupants in 143 frontal collisions with an overlap exceeding 25%, where the crash pulses have been recorded using crash pulse recorders. The results show that the shape of the crash pulse influences the risk of long-term disability to the neck. The vehicle accelerations in the mid and last third of the crash pulse seem to be important. It is also shown how change of velocity and mean and peak accelerations influence the neck-injury risk. It is suggested that the risk of sustaining an AIS1 neck injury in frontal impacts could be reduced by using more effective pretensioners and more advanced belt-load limiters. These results may also have implications for neck injury mechanisms in rear-end impacts. PMID- 10688477 TI - The relationship between clinical and kinematic responses from human subject testing in rear-end automobile collisions. AB - Recent experiments have produced a linked data set of clinical and kinematic responses for human subjects exposed to controlled low-speed rear-end collisions. The purpose of this paper was to examine this paired data set and determine whether the presence or absence of clinical symptoms could be predicted from the peak linear and angular kinematic response of the head and neck. The data were generated using 42 male and female human subjects seated normally in the front passenger seat of a stationary vehicle struck from behind to produce vehicle speed changes of 4 and 8 km/h. Pre- and post-test clinical examinations documented the presence, severity and duration of whiplash-associated disorders (WAD). Logistic regression and backward elimination of independent variables were used to develop the prediction model. The analysis yielded a 16 parameter model that was significantly related (odds ratio = 21.2; P = 0.0069) to the presence or absence of transient whiplash symptoms. The model correctly predicted symptom presence in 13 of 23 tests (sensitivity 57%) and symptom absence in 49 of 52 tests (specificity 94%) in a population of 75 with a symptom prevalence of 31%. The model's positive predictive value was 81% and its negative predictive value was 83%. Despite statistical significance, the model did not discriminate between the presence and absence of symptoms in all tests, and indicated that factors other than the selected peak kinematic responses influenced symptom production. PMID- 10688478 TI - Seat back and head restraint response during low-speed rear-end automobile collisions. AB - Automobile seat backs and head restraints play a key safety role during low-speed rear-end collisions, yet few studies have explored the effect of collision variables on seat response. In this study, the effects of vehicle speed change and seat belt use on dynamic seat back and head restraint response during low speed rear-end automobile collisions were examined. Four human subjects were repeatedly exposed to vehicle-to-vehicle rear-end collisions with speed changes of 2, 4, 6 and 8 km/h. Seat back force and deflection, and head restraint force were measured. The point of application of the resultant force applied to the seat back and head restraint were determined. The magnitude and time of peak kinematic and kinetic response parameters were used in a two-way repeated measures analysis of variance (ANOVA) for speed change and seat belt use. The results showed that 20 of the 24 seat back and head restraint response parameters varied with speed change and none of the parameters varied with seat belt use. Head restraint forces, seat back forces and seat back deflections increased approximately linearly with speed change, whereas time to peak response, direction and moment arm of the forces remained either constant or varied only slightly over the range of speed changes tested. PMID- 10688479 TI - Role of awareness in head-neck acceleration in low velocity rear-end impacts. AB - Fourteen normal healthy seated and restrained young adults were delivered rear end impacts of four intensities of acceleration. The chair was delivered a regulated and controlled pneumatic blow using a 30 cm cylinder to cause an acceleration of 0.5, 0.9, 1.1 and 1.4g. The accelerated chair was stopped suddenly by impacting the stopper at the other end of the 2 m long friction reduced track. In one set of trials, subjects were informed about the impending impact and in the other they were blindfolded and provided with loud auditory input to eliminate cues of the impact. The accelerations of the chair, shoulder and head of the participating subjects were measured triaxially and compared between levels of acceleration and expectation. The multiple analyses of variance revealed that the peak acceleration was significantly affected by the gender (P < 0.01), intensity of impact (P < 0.001), and expectation (P < 0.0001). The accelerations were significantly different in different axes (P < 0.001). A significant two-way interaction between acceleration and expectation (P < 0.03), and expectation and axes of acceleration (P < 0.02) would imply that awareness of the impending impact serves to significantly reduce the level of accelerations of head and neck. PMID- 10688480 TI - Influence of seat characteristics on occupant motion in low-speed rear impacts. AB - To analyze the effect of the seat characteristics on dummy motions and human volunteer motions, sled tests simulating low-speed rear impacts were conducted with some seats which had different characteristics. Volunteer's cervical vertebral motions were photographed with an X-ray cineradiographic system at a speed of 90 frames/s as well as the visible motions of dummy's and volunteer's were recorded. Although the tests were conducted under limited conditions, the results indicated the relationship between the occupant's visible motions, which are assumed to be closely related to the whiplash injury mechanism, and seat characteristics. It should be noted that the volunteer sled tests were discussed and approved by the Tsukuba University Ethics Committee and the volunteer submitted his informed consent in writing in line with the Helsinki Declaration. PMID- 10688481 TI - Pressure measurements in the spinal canal of post-mortem human subjects during rear-end impact and correlation of results to the neck injury criterion. AB - The aim of this study is to validate the pressure effect theory on human beings during a realistic rear-end impact and to correlate the neck injury criterion to pressure in the spinal canal. Sled experiments were performed using a test setup similar to real rear-end collisions. Test conditions were chosen based on accident statistics and recordings of real accidents. In particular, velocity change and acceleration level were reproduced similar to actual collisions. The head restraint as well as the seat back were adjusted to different positions. Two small pressure transducer were implemented to the spinal canal of postmortem human subjects and pressure measurement similar to the pig experiments (using exactly the same equipment) were performed. A total set of 21 experiments with four different subjects were performed. The subjects were additionally instrumented with triaxial accelerometers that allowed for calculation of the NIC criterion. Results showed that NIC and pressure amplitudes of the CSF correlate well and therefore NIC seems to be able to predict these amplitudes also for human beings. Conclusions whether these pressure effects induce soft tissue neck injuries or not could not be drawn and should be investigated in further research. PMID- 10688482 TI - A new mathematical neck model for a low-velocity rear-end impact dummy: evaluation of components influencing head kinematics. AB - A mathematical model of a new rear-end impact dummy neck was implemented using MADYMO. The main goal was to design a model with a human-like response of the first extension motion in the crash event. The new dummy neck was modelled as a series of rigid bodies (representing the seven cervical vertebrae and the uppermost thoracic element, T1) connected by pin joints, and supplemented by two muscle substitutes. The joints had non-linear stiffness characteristics and the muscle elements possessed both elastic stiffness and damping properties. The new model was compared with two neck models with the same number of vertebrae, but without muscle substitutes. The properties of the muscle substitutes and the need of these were evaluated by using three different modified neck models. The motion of T1 in the simulations was prescribed using displacement data obtained from volunteer tests. In a sensitivity analysis of the mathematical model the influence of different factors on the head-neck kinematics was evaluated. The neck model was validated against kinematics data from volunteer tests: linear displacement, angular displacement, and acceleration of the head relative to the upper torso at 7 km/h velocity change. The response of the new model was within the corridor of the volunteer tests for the main part of the time history plot. This study showed that a combination of elastic stiffness and damping in the muscle substitutes, together with a non-linear joint stiffness, resulted in a head-neck response similar to human volunteers, and superior to that of other tested neck models. PMID- 10688483 TI - Road safety engineering: an effective tool in the fight against whiplash injuries. AB - Road safety engineering can play an integral part in the prevention of whiplash injuries. While improvements to vehicle design can reduce the severity of whiplash injuries when a crash occurs, improvements to road safety can prevent whiplash-inducing crashes from occurring in the first place. Whiplash injuries are most commonly associated with rear end crashes. Unfortunately, rear end crashes are also the most common type of crash at urban signalized intersections, where the majority of crashes occur in British Columbia, Canada. The Insurance Corporation of British Columbia (ICBC), through the road improvement program, has been funding road improvements in order to reduce the frequency of collisions at high crash locations in British Columbia. Several road safety engineering countermeasures specifically targeted at rear end collisions have been researched and deployed. These countermeasures include simple and affordable solutions such as signal visibility enhancements, as well as complex and expensive solutions such as intersection geometric upgrades. When appropriately used, these countermeasures have proven to be extremely cost-effective in reducing the frequency of rear end collisions. Widespread application of signal visibility enhancements is now being pursued to further decrease the risk of rear end collisions and whiplash injuries. Costs are the direct cost of the ICBC portion of the investment and benefits are only those associated with reduced insurance claims over a 2-year period. PMID- 10688484 TI - Injury surveillance in Victoria, Australia: developing comprehensive injury incidence estimates. AB - This study aimed to develop an estimate of the incidence of all medically-treated injury by level of severity and to broadly describe the epidemiology of injury in the Australian State of Victoria in a given year. Victoria has developed a relatively comprehensive injury surveillance system. Data is currently collected by various agencies on injury deaths, hospitalisations and emergency department attendances. The method used to establish the incidence of both unintentional and intentional injury is described. Incidence figures were directly derived, or estimated from, the available Victorian health sector and Coronial data bases for three level of severity (deaths, hospitalisations and medical treatment only) and for causes of injury, age and gender groups, location of the injury event and activity at the time of injury. In 1993/1994, injuries resulted in at least 1487 deaths, 67,402 persons hospitalised and an estimated 397,160 medically-treated, non-hospitalised injured persons in Victoria. In total, over 466,000 people were injured or 10.5 persons per year for every 100 residents. Males sustain 62% of all injuries yet represent 49.5% of the population. Almost three-quarters of injury fatalities and over 60% of non-fatal injuries occur among males. Young people aged 15-24 years account for 22% of all injuries yet represent only about 16% of the Victorian population. Children (0-14 years) also suffer relatively high injury rates, although mainly less severe, while the elderly are at risk of more severe injuries. The leading cause of injury death in Victoria is suicide, followed by motor vehicle accidents, whereas falls are the leading cause of all non-fatal injury. Most injuries occur in the home (36%), areas of sport and recreation (12.5%) and transport (11.7%). They are mainly associated with leisure activities (33.1%), work (11%) and transportation (10.8%). This study demonstrates a method for the development of comprehensive injury incidence estimates. The results indicate that injuries have a significant impact on the Victorian community, health care system and economy in general. Reliable incidence data are necessary for descriptive epidemiology and provide the basis for quality of life and economic cost studies. Together this information has potential application for evidence-based strategic planning and evaluation in injury research and prevention. PMID- 10688485 TI - Neck pain and head restraint position relative to the driver's head in rear-end collisions. AB - This two-year investigation was designed to estimate the incidence of driver neck pain in rear-struck vehicles involved in two-vehicle collisions and to determine the relationship between neck pain and specific vehicle, human, and environmental factors. Neck pain percentages were significantly higher for female (45%) than for male (28%) drivers. For female and male drivers, neck pain likelihood increased as head restraint height decreased below the head's center of gravity, although this effect was significant only for females. Head restraint backset, the horizontal distance measured from the back of the driver's head to the front of the head restraint, was not found to be related to neck pain for female drivers. Backset trends for male drivers could not be evaluated because few male drivers had head restraints that were high enough for backset to be relevant. Reported neck pain decreased for older drivers (females only), drivers in less severe crashes, and drivers in heavier cars (females only); all head restraint analyses were adjusted for these characteristics. Women, and most likely men, in the United States would benefit greatly from international harmonization to European head restraint standards. Until then, both women and men should be encouraged to adjust their adjustable head restraints, if possible, behind their heads' centers of gravity and to sit with the backs of their heads as close as possible to their head restraints. PMID- 10688486 TI - Report investigating the importance of head restraint positioning in reducing neck injury in rear impact. AB - Neck injury resulting from rear impact (often known as whiplash) is a serious cause of road trauma. It is often underestimated or overlooked because such injuries are minor on traditional injury scales but can result in long term pain and disability. The paper begins with a brief review of research into head restraints and whiplash done so far. A review of international head restraint regulations revealed the absence of any horizontal offset requirements. A review of seat strength requirements and testing procedures showed that a regulation that required a collapsible seat would involve significant compliance testing. This paper concludes a preliminary project conducted by the Federal Office of Road Safety (FORS) where the head restraints for twenty Australian market vehicles were assessed using known performance criteria. A key finding of the report was that most of the vehicles allowed for vertical adjustment of the head restraint. Also important was that none of the vehicles measured allowed horizontal adjustment and on some of the head restraints the horizontal displacement increased as the vertical height increased. As the understanding of neck injury mechanisms in rear impact develops, there may be some scope for FORS to facilitate the improvement of these standards. Further research into neck injury mechanisms may reveal yielding seat backs or new 'active' head restraint technology as a more effective countermeasure. In the meantime, educating occupants to correctly adjust their head restraints seems to be an effective way to reduce injuries in existing vehicles. PMID- 10688487 TI - WHIPS--Volvo's Whiplash Protection Study. AB - Whiplash associated disorders (WAD) resulting from rear end car impacts are an increasing problem. WAD are usually not life threatening, but are one of the most important injury categories with regard to long-term consequences. This paper is a review of Volvo's Whiplash Protection Study (WHIPS), which is the result of more than ten years of concentrated research efforts in the area of neck injuries in car collisions, with the focus on rear end car impacts. The study follows the whole chain from accident research to the development of a seat for increased protection against WAD. Results from Volvo's accident research are summarized. Existing biomechanical knowledge regarding possible injury mechanisms are presented and discussed. Based on the interpretation of accident research and biomechanical knowledge, guidelines for improved protection against WAD in rear end impacts are presented. Requirements and test methods based on the guidelines are explained. An important part of the study is a new rear end impact dummy, BioRID. Test results using the new dummy are presented. Finally, the paper explains the design of a new seat for increased WAD protection, the WHIPS-seat. Results from the accident research and biomechanical research emphasize the importance of considering the whole spine of the occupant and, accordingly, the whole seat when addressing WAD in rear end impacts, with a particular focus on low and moderate impact severity. Low and moderate impact severity crashes should be focused. Also important to consider are the individual differences between occupants, the seating position and the variety of seating postures. All results, including sub-system testing, mathematical modeling, sled testing, as well as geometrical parameters show that the WHIPS-seat will have considerable potential for offering increased protection against WAD in rear end impacts. PMID- 10688488 TI - Comparison of car seats in low speed rear-end impacts using the BioRID dummy and the new neck injury criterion (NIC). AB - Long-term whiplash associated disorders (WAD) 1-3 sustained in low velocity rear end impacts is the most common disability injury in Sweden. Therefore, to determine neck injury mechanisms and develop methods to measure neck-injury related parameters are of importance for current crash-safety research. A new neck injury criterion (NIC) has previously been proposed and evaluated by means of dummy, human and mathematical rear-impact simulations. So far, the criterion appears to be sensitive to the major car and collision related risk factors for injuries with long-term consequences. To further evaluate the applicability of NIC, four seats were tested according to a recently proposed sled-test procedure. 'Good' as well as 'bad' seats were chosen on the basis of a recently presented disability risk ranking list. The dummy used in the current tests was the Biofidelic Rear Impact Dummy (BioRID). The results of this study showed that NICmax values were generally related to the real-world risk of long-term WAD 1-3. Furthermore, these results suggested that NICmax calculated from sled tests using the BioRID dummy can be used for evaluating the neck injury risk of different car seats. PMID- 10688489 TI - Intrapleural therapy with MDP-Lys (L18), a synthetic derivative of muramyl dipeptide, against malignant pleurisy associated with lung cancer. AB - N2-[(N-acetylmuramoyl)-L-alanyl-D-isoglutaminyl]-N6-stearoyl-L-lysine (MDP-Lys (L18), romurtide) is a synthetic muramyl dipeptide derivative, and has immunomodulating activities including activation of cells of monocyte-macrophage lineage. We examined the effect of intrapleural instillation of MDP-Lys (L18) against malignant pleurisy associated with lung cancer. Six patients with cytologically-positive malignant pleural effusion (four with adenocarcinoma, one with small cell carcinoma and one with large cell carcinoma) were treated with single intrapleural instillation of MDP-Lys (L18) of 200 microg. Clinically, no reaccumulation of pleural effusion for at least 4 weeks was observed in four patients. No major side effects were observed. Total cell number elevated remarkably 4 h after instillation, and main increased population was that of neutrophils. Levels of chemotactic cytokines, such as interleukin (IL)-8, and monocyte chemotactic protein (MCP)-1 and levels of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6, elevated in pleural effusion, and peak IL-1beta and IL-6 levels tended to be higher in clinical responders than non-responders. These results suggest MDP-Lys (L18) instilled by intrapleural route had a potential local immunomodulatory activity. Further study is warranted to further determine the critical factors which correlate with the clinical response. PMID- 10688490 TI - Gemcitabine monotherapy in elderly patients with advanced non-small cell lung cancer: a multicenter phase II study. AB - BACKGROUND: This trial investigated the activity and toxicity of gemcitabine in previously untreated elderly (> 70 years) patients with advanced (stage IIIB-IV) non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: From January 1997 to July 1998, 46 patients with advanced NSCLC aged over 70 years with a performance status of 0-2 were entered into the study. Gemcitabine 1000 mg/m2 was administered as a 30-min infusion once a week for 3 weeks followed by a week of rest; cycles were repeated every 4 weeks. RESULTS: Forty-four patients were evaluable for response. One complete response and nine partial responses were observed, for an overall response rate of 22.2% (95% C.I.: 11.3-37.5). The median time to disease progression was 5.1 months (95% C.I.: 3.5-6.7), the median duration of response was 6.3 months, and the median overall survival time 6.75 months (95% C.I.: 5.3-8.2). All patients were evaluable for toxicity (184 cycles, median = 3 cycles/patient) and no grade 4 hematologic toxicities were reported. WHO grade 3 leukopenia, neutropenia and anemia occurred in 3.3, 0.5 and 1.1% of cycles, respectively. Grade 3 skin rash occurred in 4.3% of patients. These side effects led to treatment discontinuation in two patients. CONCLUSION: Our data show that gemcitabine is active and well tolerated in patients aged over 70 years with advanced NSCLC. PMID- 10688491 TI - Angiogenesis and non-small cell lung cancer. AB - Lung cancer is the commonest cause of cancer death in the western world. Recent evidence suggests that angiogenesis is related to poor prognosis in many solid tumours including non-small cell lung cancer. Angiogenesis is controlled by a complex interaction between growth and apoptotic factors, proteases and adhesion molecules. The angiogenic process may prove a target for novel therapies such as matrix metalloproteinase inhibitors, growth factor antisense RNA, growth factor receptor antagonists and naturally occurring antiangiogenic peptides. These agents may be used alone or in combination with traditional chemotherapy, radiotherapy and surgery. PMID- 10688492 TI - Surgical treatment of lung cancer with adrenal metastasis. AB - Surgical treatment of adrenal metastasis from non-small cell lung cancer is controversial. Classically this group of patients has been considered incurable, therefore excision of the primary cancer and the adrenal gland has been avoided. However, recent reports show good results in their surgical management. Five selected patients with non-small cell lung cancer and adrenal metastases have been surgically treated. Two of them also presented with brain metastases that were excised, too. One patient with brain and adrenal metastases died 38 months after surgery. The other four patients are alive and with no sign of recurrent disease at 8, 16, 52 and 58 months of follow-up. In highly selected patients in whom both the primary and the metastatic tumors are resectable and in the absence of tumor spread to other organs, surgical treatment seems to be a good therapeutic option. PMID- 10688493 TI - Incidence of lung cancer in Denmark: historical and actual status. AB - This is a registry-based analysis of 97,281 lung cancer patients diagnosed in Denmark during the period 1943-1994. The development of lung cancer incidence in Denmark in the study period is described and this information is used to predict the future trends in lung cancer in Denmark. Since 1960, lung cancer has been the most frequent type of cancer in Danish men, excluding skin cancer. The incidence in men reached its maximum in 1985, when it was 100 new cases per 100,000 inhabitants, whereafter the incidence began to level off and was 83 per 100,000 inhabitants in 1994. During the study period, the incidence of lung cancer in Danish women was somewhat lower than in men. The incidence in women has been steadily increasing since 1960 by approximately 20% per each 5-year period, and was 46 per 100,000 inhabitants in 1994. Adenocarcinoma is the most frequent histological subtype in women and the only subtype increasing in incidence in men. Taking the actual development into account, the incidence of lung cancer will continue to fall among men and rise among women, leading to a reversal of the classical gender ratio in about 15 years. PMID- 10688494 TI - Early hilar lung cancer--clinical aspects and long term survival. Identification of a subgroup of stage IA patients with more favorable prognosis. AB - Twenty-nine patients out of 2018 operated on for a non-small-cell lung cancer from 1987 to February 1998 met the criteria proposed by the Japan Lung Cancer Society (JLCS) for the definition of early hilar lung cancer (EHLC). Twenty-six patients were symptomatic and 20 had a radiologically visible lesion. All cancers were located and diagnosed by bronchoscopy and all patients were resected. At histology, all tumors were squamous in nature. The five-year cumulative survival rate was 96%--a second primary lung cancer (2nd Pr.) developed in 4 patients (13.8%). The definition of EHLC proposed by the JLCS allows the selection of a subgroup of stage I patients with a very good prognosis. Nevertheless, a close follow-up is mandatory because more than 10% of these patients develop a 2nd Pr. PMID- 10688495 TI - Synchronous four primary lung adenocarcinoma associated with multiple atypical adenomatous hyperplasia. AB - A 69-year-old woman with synchronous bilateral 4 primary lung adenocarcinoma accompanied by multiple atypical adenomatous hyperplasia (AAH) is described. The patient was found to have bilateral multiple tumors during a preoperative chest CT evaluation which was performed for the previously-diagnosed adenocarcinoma of the right middle lobe. Since intraoperative diagnosis of the left nodular lesion was adenocarcinoma and judged to be a pulmonary metastasis, a lobectomy of the right middle lobe only was performed. Postoperative pathological diagnosis including immunohistochemical findings demonstrated that the bilateral lesions were synchronous multiple primary adenocarcinoma, independent of each other and associated with multiple AAH. This case suggests the possibility of the AAH adenocarcinoma sequence in the development of lung cancer. In addition, the strategy of treatment for synchronous multiple adenocarcinoma should be considered. PMID- 10688496 TI - Sling techniques in the treatment of genuine stress incontinence. PMID- 10688497 TI - Altered peripheral vascular response of women with and without pelvic pain due to congestion. AB - OBJECTIVE: To test the hypothesis that women with pelvic venous congestion have a reduction of reactivity of their peripheral circulation. DESIGN: Comparison was made between 20 women with chronic pelvic pain due to congestion and a control group of 15 pain-free women matched for age, parity and body weight. A comparison of these results was made with those from six postmenopausal women taking hormone replacement therapy. METHODS: Study and control groups were investigated during the mid-follicular phase of the menstrual cycle (days 5-9) and the mid-luteal phase (days 19-23). The study group was also investigated during the fifth month of treatment with suppression of ovarian activity with leuprorelin or medroxyprogesterone acetate or six months after hysterectomy and bilateral salpingo-oophorectomy. Head-up tilt sufficient to increase intra-vascular pressure in the toe by a standard 40 mmHg was used as a means of raising venous pressure in the lower limb. Skin capillary red blood cell velocity (flux) was measured using a laser Doppler flow probe placed over the pulp of the big toe. Heart rate and blood pressure were also recorded. The change in skin blood flow following head-up tilt was expressed as a percentage of baseline flow in the supine position. MAIN OUTCOME MEASURES: Percentage change in skin red blood cell flux, heart rate and blood pressure in response to 40 degrees head-up tilt. RESULTS: In the control group the median response to head-up tilt in the follicular phase was one of a reduction in flux, whereas in the luteal phase it was more variable ranging from an increase to a decrease in flux. The responses in the pelvic congestion group in both the follicular and luteal phases were similar to those of the control group in the luteal phase. A small but significant increase in heart rate in response to tilt in the pelvic pain group, compared with the control group, was interpreted as being due to a fall in venous return. Treatment of the pelvic congestion group by medical suppression of ovarian activity or total hysterectomy with bilateral salpingo-oophorectomy resulted in a significant change in response to head-up tilt from the variable type of luteal response to one of a more constant reduction in flux, similar to that of the control group in the follicular phase. A reduction in flux was also found consistently in postmenopausal women. CONCLUSION: The study confirms the hypothesis that women with pelvic pain due to congestion show a change in peripheral vascular reactivity which returns to normal after suppression of ovarian activity. It seems likely that some alteration of normal ovarian function is responsible for the observed changes in peripheral blood flow in response to a rise in venous pressure in women with pelvic congestion. PMID- 10688498 TI - Performance of ultrasound as a second line test to serum CA125 in ovarian cancer screening. AB - OBJECTIVE: To assess the performance of ultrasonography in a multimodal ovarian cancer screening strategy. DESIGN: Prospective ovarian cancer screening trial between December 1986 and June 1993. SETTING: General practice, occupational health departments and an ovarian cancer screening clinic at a London teaching hospital. POPULATION: Postmenopausal women, > or = 45 years with a raised CA125. METHODS: Volunteers with a CA125 > or = 30 U/mL underwent a pelvic ultrasound. Scans were classified as normal, abnormal (ovarian volume > or = 8.8 mL) or equivocal (normal volume with abnormal morphology). Abnormal ovarian morphology was subclassified as simple cyst (single, thin walled cyst with no septa or papillary projections) or complex (all other abnormalities). Volunteers with abnormal scans were referred for a gynaecological opinion. Follow up was via the cancer registry and postal questionnaires. MAIN OUTCOME MEASURES: Sensitivity, specificity and positive predictive value of different ultrasound criteria for detection of index cancer (e.g. primary invasive epithelial carcinoma of the ovary and fallopian tube). RESULTS: Seven hundred and forty-one women underwent 1,219 scans and 20 index cancers occurred during a median follow up of 6 x 8 years. The sensitivity for detection of ovarian cancer of different ultrasound criteria was 100% for abnormal morphology, 89 x 5% for abnormal volume and 84% for complex morphology. The highest specificity (97%) and positive predictive value (37 x 2%) was achieved using complex morphology. CONCLUSION: A variety of ultrasound criteria can achieve high sensitivity, specificity and positive predictive value for index cancers in postmenopausal women with an elevated CA125. Use of ovarian morphology to interpret ultrasound may increase sensitivity and use of complex ovarian morphology may increase the positive predictive value. PMID- 10688499 TI - Gynaecological presentation of retroperitoneal tumours. AB - OBJECTIVE: To illustrate the problems associated with mistaken pre-operative diagnosis following gynaecological presentation of patients with retroperitoneal tumours. DESIGN: A case series of five referrals. RESULTS: Non-gynaecological tumours were not suspected in each case and hence there was a failure to undertake further pre-operative investigation and referral to a specialised soft tissue sarcoma service. This resulted in four of the patients having an unnecessary laparotomy with an inappropriate transperitoneal biopsy undertaken when the retroperitoneal tumour was discovered. The mistaken diagnosis of ovarian malignancy lead to increased morbidity, compromise of potential for a long disease free interval and/or possibly lessened the chance of cure in each case. CONCLUSIONS: Misinterpretation of clinical signs and an over-reliance on ultrasound diagnosis were the commonest causes of inappropriate management of these patients. Gynaecologists should consider more frequently the other, less common differential diagnoses of a pelvic mass. This is especially true in circumstances with a predominantly solid tumour, where there are clinical signs of vascular or rectal displacement, or where there is ultrasound evidence of ureteric obstruction. The more frequent utilisation of a computerised tomography scan with intravenous and oral contrast with referral before inappropriate transperitoneal biopsy are recommended as complete en bloc surgical excision at the first laparotomy is the treatment of choice in virtually all primary retroperitoneal tumours. PMID- 10688500 TI - Can diagnostic laparoscopy be avoided in routine investigation for infertility? AB - OBJECTIVE: To determine whether routine testing for serum Chlamydia trachomatis antibodies, considered in combination with a woman's clinical features, may avoid the need for diagnostic laparoscopy in routine investigation for infertility. DESIGN: Retrospective case notes analysis. SETTING: Secondary level care infertility clinic. POPULATION: Eighty women who had undergone both laparoscopy and serum Chlamydia trachomatis antibody testing. METHODS: Ascertainment of any history of suspected pelvic inflammatory disease, pelvic pain, cervical intraepithelial neoplasia, pelvic surgery or appendicectomy; any abnormality on clinical pelvic examination; the findings at laparoscopy; the result of serum Chlamydia trachomatis antibody testing by enzyme-linked immunosorbent assay (ELISA) screening with microimmunofluorescence (MIF) confirmatory diagnostic testing. The usefulness of clinical features, the serum Chlamydia trachomatis antibody test and these two variables combined in the detection of tubal disease and pelvic pathology of relevance to infertility were measured statistically. MAIN OUTCOME MEASURES: Specificity, sensitivity, positive predictive value, negative predictive value and likelihood ratio for each of the tests. RESULTS: The combination of any positive clinical feature with a positive test for serum Chlamydia trachomatis antibodies detects tubal disease with sensitivity 92%, specificity 70%, positive predictive value 72%, negative predictive value 91% and likelihood ratio 3 x 1; it detects bilateral tubal obstruction with sensitivity 84%, specificity 51%, positive predictive value 35%, negative predictive value 91% and likelihood ratio 1 x 7; it detects pelvic pathology relevant to infertility with sensitivity 76%, specificity 71%, positive predictive value 80%, negative predictive value 65% and likelihood ratio 2 x 6. The negative predictive value for pelvic pathology from the use of clinical features in addition to the chlamydia antibody test is not significantly higher than that from the chlamydia antibody test alone (53%). CONCLUSIONS: A policy of selective laparoscopy in routine investigation for infertility, based on the result of the test for serum Chlamydia trachomatis antibodies and a woman's clinical features, is not supported. PMID- 10688501 TI - Births in Finland and Estonia from 1992 to 1996: convergent differences? AB - OBJECTIVE: To describe the differences in childbearing, in prenatal and obstetrical practices, and in perinatal health outcome in Finland and Estonia. DESIGN: Registry study using the data from the Finnish and Estonian medical birth registries for years 1992 to 1996 (in total 324,021 and 74,297 newborns, respectively). RESULTS: In 1992 the birth rates were 51 per 1,000 women aged 15 to 49 in Finland and 48 per 1,000 in Estonia. The birth rate declined in the study period in both countries, but the decline was more rapid in Estonia (-26%) than in Finland (-6%). In the same period the rates of induced abortion declined in both countries (-34% and -6%, respectively), but the rate in 1996 was still much higher in Estonia (46/1,000) than in Finland (8/1,000). Compared with Finnish mothers, Estonian mothers were younger, had fewer multiple births, less prenatal care and fewer interventions during pregnancy and delivery. The intervention rates increased in both countries during the study period, but this increase was more rapid in Estonia. The infant outcomes were poorer in Estonia, but the differences between Estonia and Finland decreased during the 1990's. CONCLUSIONS: The differences in prenatal and maternal care and in induced abortion rates have decreased between Estonia and Finland. Changes in maternal backgrounds, improved referral system for complicated pregnancies, improvements in prenatal care and in availability of appropriate equipment and technology may have caused improved maternal and infant health in Estonia, but this should be further investigated. PMID- 10688502 TI - Women's sexual health after childbirth. AB - OBJECTIVE: To investigate the impact of childbirth on the sexual health of primiparous women and identify factors associated with dyspareunia. DESIGN: Cross sectional study using obstetric records, and postal survey six months after delivery. SETTING: Department of Obstetrics and Gynaecology, St George's Hospital, London. POPULATION: All primiparous women (n = 796) delivered of a live birth in a six month period. METHODS: Quantitative analysis of obstetric and survey data. MAIN OUTCOME MEASURES: Self reported sexual behaviour and sexual problems (e.g. vaginal dryness, painful penetration, pain during sexual intercourse, pain on orgasm, vaginal tightness, vaginal looseness, bleeding/irritation after sex, and loss of sexual desire); consultation for postnatal sexual problems. RESULTS: Of the 484 respondents (61% response rate), 89% had resumed sexual activity within six months of the birth. Sexual morbidity increased significantly after the birth: in the first three months after delivery 83% of women experienced sexual problems, declining to 64% at six months, although not reaching pre-pregnancy levels of 38% . Dyspareunia in the first three months after delivery was, after adjustment, significantly associated with vaginal deliveries (P = 0 x 01) and previous experience of dyspareunia (P = 0 x 03). At six months the association with type of delivery was not significant (P = 0 x 4); only experience of dyspareunia before pregnancy (P < 0 x 0001) and current breastfeeding were significant (P = 0 x 0006). Only 15% of women who had a postnatal sexual problem reported discussing it with a health professional. CONCLUSIONS: Sexual health problems were very common after childbirth, suggesting potentially high levels of unmet need. PMID- 10688503 TI - How useful is uterine artery Doppler flow velocimetry in the prediction of pre eclampsia, intrauterine growth retardation and perinatal death? An overview. AB - OBJECTIVE: To evaluate the clinical usefulness of Doppler analysis of the uterine artery velocity waveform in the prediction of pre-eclampsia and its associated complications of intrauterine growth retardation and perinatal death. DESIGN: Quantitative systematic review of observational diagnostic studies using online searching of the MEDLINE database coupled with scanning of the bibliographies of primary and review articles including known unpublished studies. MATERIAL: Twenty seven studies involving 12,994 subjects stratified into population subgroups at low and high risk of developing pre-eclampsia and its complications. OUTCOME MEASURES: The outcome measures studied were: 1. the development of pre-eclampsia; 2. intrauterine growth retardation; and 3. perinatal death. The main meta analyses were the flow velocity waveform ratio +/- diastolic notch derived by transabdominal Doppler ultrasound as the measurement parameter. The analyses were conducted using likelihood ratio as a measure of diagnostic accuracy. A likelihood ratio of 1 indicates that the test has no predictive value for the outcome. Prediction for the outcome event is considered conclusive with likelihood ratios of > 10 or < 0 x 1 for a positive and negative test result, respectively. Moderate prediction can be achieved with likelihood ratios of 5-10 and 0 x 1-0 x 2 whereas likelihood ratios values of 1-5 and 0 x 2-1 would generate only minimal prediction. RESULTS: In the low risk population a positive test result, predicted pre-eclampsia with a pooled likelihood ratio of 6 x 4 (95% CI 5 x 7-7 x 1), while a negative test result had a pooled likelihood ratio of 0 x 7 (95% CI 0 x 6-0 x 8). For intrauterine growth retardation the pooled likelihood ratio was 3 x 6 (95% CI 3 x 2-4 x 0) for a positive test result and 0 x 8 (95% CI 0 x 8-0 x 9) for a negative test result. Using perinatal death as outcome measure, the pooled likelihood ratio was 1 x 8 (95% CI 1 x 2-2 x 9) for a positive test result and 0 x 9 (95% CI 0 x 8-1 x 1) for a negative test result. In the high risk population a positive test result predicted pre-eclampsia with a pooled likelihood ratio of 2 x 8 (95% CI 2 x 3-3 x 4), while a negative test had a likelihood ratio of 0 x 8 (95% CI 0 x 7-0 x 9). For intrauterine growth retardation the pooled likelihood ratio was 2 x 7 (95% CI 2 x 1-3 x 4) for a positive test result and 0 x 7 (95% CI 0 x 6-0 x 9) for a negative result. For perinatal death the pooled likelihood ratio was 4 x 0 (95% CI 2 x 4-6 x 6) for a positive test result and 0 x 6 (95% CI 0 x 4-0 x 9) for a negative result. CONCLUSION: Uterine artery Doppler flow velocity has limited diagnostic accuracy in predicting pre-eclampsia, intrauterine growth retardation and perinatal death. PMID- 10688504 TI - Introduction of the Misgav Ladach caesarean section at an African tertiary centre: a randomised controlled trial. AB - OBJECTIVE: To determine whether the Misgav Ladach caesarean section technique can offer benefits compared with conventional caesarean section technique in the prevailing conditions of a busy African tertiary centre. DESIGN: A randomised controlled trial. SETTING: A tertiary African obstetric unit with 18,000 deliveries annually. PARTICIPANTS: Three hundred and thirty-nine women undergoing caesarean section. METHODS: Eight residents and registrars were instructed in the Misgav Ladach technique for caesarean section during one week, after which the study commenced. The course participants instructed their colleagues; in total, 16 doctors participated. Women requiring caesarean section were randomised to Misgav Ladach or to the conventional lower midline incision procedure, excluding those with a previous scar. RESULTS: During 11 weeks 339 randomised procedures (328 of which were emergency procedures) were carried out. Mean operating time was 25 x 3 minutes for Misgav Ladach and 32 x 6 minutes for the lower midline incision procedure (95% CI -8 x 3; -6 x 3). Mean blood loss was 354 mL and 447 mL (-133; -53), and the number of sutures 3 x 1 and 6 x 1 (-3 x 1; -2 x 9), respectively. No significant difference was found in Apgar scores. Mobilisation was earlier with the Misgav Ladach procedure. No difference was found in overall post-operative infection rates i.e. wound infection or febrile illness, but the combination of wound infection and fever was more common in the Misgav Ladach group. CONCLUSION: The Misgav Ladach caesarean section confers benefits such as reduced blood loss, conservation of time and suture material, and rapid mobilisation, but more studies are needed to explore modifications aimed at reducing post-operative infections in settings with limited resources. PMID- 10688505 TI - Antenatal home blood pressure monitoring: a pilot randomised controlled trial. AB - OBJECTIVE: To measure recruitment to, compliance with, and the acceptability of a trial designed to test whether a reduced schedule of antenatal visits combined with training in self-measurement of blood pressure at home may improve hypertension screening and save money. To test the specific hypothesis that even after taking into account extra unscheduled visits, the reduced schedule with ambulatory monitoring reduces total visits. DESIGN: A pilot randomised controlled trial. SETTING: Four urban and four rural general practices in Yorkshire and Lancashire. POPULATION: One hundred and five low risk women in the third trimester of pregnancy. Eighty women participated. INTERVENTION: Women were invited to participate at 24-28 weeks. Those who accepted were allocated either to a standard nine subsequent visit schedule (30, 32, 34, 36, 37, 38, 39, 40, 41 weeks) or to a reduced schedule (34, 38, 41 weeks). Those in the latter group measured their blood pressure weekly using a portable sphygmomanometer at home. MAIN OUTCOME MEASURES: Recruitment, total number of clinic visits, frequency of blood pressure measurement, schedule preference, and anxiety. RESULTS: Although there were more unscheduled visits in the home monitoring group, this did not outweigh the reduction in scheduled visits, (total visits reduced from 7 x 4 to 4 x 5, P < 0 x 001), and blood pressure was measured during more weeks (9 vs 7 weeks, P < 0 x 001) in the experimental group. Most women expressed a preference for the reduced schedule both when the idea was first suggested, and after they had experienced it, and there were no significant differences in anxiety. CONCLUSION: Replacement of antenatal screening visits with home blood pressure monitoring is acceptable to women. The reduction in clinic visits is not compensated by an increase in visits for other reasons and overall blood pressure measurement is omitted less often. Whether it reduces adverse outcomes or has any rare side effects will require a larger trial, but this pilot study indicates that it is likely to be safe, and that such a large trial would be feasible. PMID- 10688506 TI - Labour characteristics and uterine activity: misoprostol compared with oxytocin in women at term with prelabour rupture of the membranes. AB - OBJECTIVE: To compare the labour pattern and uterine activity of oral misoprostol with oxytocin for labour induction in women presenting with prelabour rupture of membranes at term. DESIGN: Prospective randomised study. SETTING: Department of Obstetrics and Gynaecology, Queen Mary Hospital, Hong Kong. PARTICIPANTS: Eighty women presenting with prelabour rupture of membranes at term. METHODS: The women were randomised to receive either 100 microg misoprostol orally every 4 hours to a maximum of three doses, or intravenous oxytocin infusion according to the hospital protocol. Intrauterine pressure transducers were inserted one hour before induction of labour in both groups of women. We compared the pattern of uterine activity, the induction-to-delivery interval, duration of labour, mode of delivery and neonatal outcome between the two groups. RESULTS: Both oxytocin and oral misoprostol caused an increase in uterine activity within one hour of labour induction. Peak uterine activity was reached 6-8 h after oral misoprostol, with persistent effects, and 8-10 h after oxytocin, requiring continuous titration of medication. The duration of labour was significantly reduced in nulliparous women, but not in those who were multiparous in the misoprostol group. The induction-to-delivery interval, the mode of delivery and the perinatal outcome were similar for the two groups. CONCLUSION: Oral misoprostol caused earlier peak uterine activity, compared with oxytocin (6-8 h vs 8-10 h). Oral misoprostol was not only as effective as oxytocin in inducing labour in women at term with prelabour rupture of the membranes, but it reduced significantly the duration of labour in nulliparous women. PMID- 10688507 TI - Early pregnancy predictors of preterm birth: the role of a prolonged menstruation conception interval. AB - OBJECTIVE: To study early pregnancy characteristics as possible risk factors associated with preterm birth. DESIGN: Retrospective analysis of prospectively collected maternity data. POPULATION: 21,069 singleton deliveries with record of a specified last menstrual period and a midtrimester dating scan. SETTING: Catchment area of tertiary centre serving a general maternity population. METHODS: Univariate and multivariate analysis. Variables included: maternal age; height; weight at first visit; parity; ethnic group; cigarette smoking and alcohol consumption recorded in early pregnancy; history of abortion; history of preterm birth; and discrepancy between menstrual dates and ultrasound dates. MAIN OUTCOME MEASURES: Adjusted odds ratios for factors associated with preterm birth, stratified according to parity (nulliparae vs multiparae) and gestational age (early preterm, 24-33 weeks; late preterm, 34-36 weeks; all preterm, < 37 weeks). Population attributable risk (aetiologic fraction) of the significant variables for preterm birth. RESULTS: The overall preterm (< 37 weeks) delivery rate according to scan dates was 7 x 0%. Preterm birth was associated with young (< 20 years), short (< or = 155 cm) and underweight (< or = 52 kg) mothers, non Europeans, cigarette smokers, previous abortion or previous preterm delivery, and a prolonged menstruation-conception interval. Preterm births which followed the spontaneous onset of labour (72%) had results which were similar to the overall group, while there were too few iatrogenic preterm deliveries for separate analysis. Logistic regression showed that associations varied in different parity and gestational age groups. For nulliparae, smoking was not associated with preterm birth, but it was strongly associated with multiparous women (adjusted OR 1 x 8, 95% CI 1 x 6-2 x 1). A past history of premature delivery had the highest risk for birth before 34 weeks in the index pregnancy (adjusted OR 5 x 1, 95% CI 3 x 4-7 x 6). A discrepancy between menstrual and scan dates of greater than +7 days, suggestive of a prolonged interval between last menstruation and conception, was present in 23 x 3% of all pregnancies, and was associated with an increased risk of preterm delivery in all gestational age categories for nulliparae (adjusted OR 1 x 5, 95% CI 1 x 3-1 x 8) and multiparae (adjusted OR 1 x 9, 95% CI 1 x 6-2 x 2). Because of its high prevalence, this variable constituted a relatively high population-attributable risk for premature birth for both nulliparae (10 x 7%) and multiparae (16 x 6%). CONCLUSIONS: A discrepancy of more than +7 days between menstrual and scan dates, indicating a prolonged interval between last menstruation and conception, is a significant predictor of preterm birth. This effect is independent of other factors such as maternal age, height, weight and smoking which are also associated with prematurity. In a maternity population with ultrasound scan dates and recorded last menstrual period, this variable can be easily calculated and used as a marker for increased surveillance. PMID- 10688508 TI - Does a discrepancy between gestational age determined by biparietal diameter and last menstrual period sometimes signify early intrauterine growth retardation? AB - OBJECTIVE: To assess the association between gestational age estimated from the last menstrual period (GA(LMP)) or from the biparietal diameter (GA(BPD)), and the subsequent birthweight for gestational age. DESIGN: Population-based follow up study. SETTING: Of 21,936 pregnancies contained in the ultrasound database, 16,387 singleton pregnancies with a reliable last menstrual period date and an ultrasound examination between 12 and 22 weeks of gestation were included. MAIN OUTCOME MEASURES: Correlation between: 1) birthweight deviation (birthweight expected weight for gestation); 2) birthweight; and 3) pregnancy length and (GA(LMP)-GA(BPD)). Relative risk of birthweight < 2,500 g and low birthweight for gestation (> 22% below normal weight) related to five levels of discrepancy between (GA(LMP)-GA(BPD)). RESULTS: (GA(LMP)-GA(BPD)) was not associated with deviation of birthweight related to GA(BPD). However the risk of low birthweight (< 2,500 g) and low birthweight for gestational age was significantly increased when (GA(LMP)-GA(BPD)) was > 7 days. CONCLUSION: A biparietal diameter smaller than expected from the last menstrual period date is mainly a problem of an error related to estimated time of ovulation. At the same time the relative risk of a low birthweight infant is slightly increased. PMID- 10688509 TI - Pregnancy outcomes and cardiac complications in women with mechanical, bioprosthetic and homograft valves. AB - OBJECTIVES: Firstly, to compare pregnancy outcomes and cardiac complications in women with: 1) either mechanical or bioprosthetic valves at the mitral site; 2) mechanical valves treated with warfarin or subcutaneous heparin. Secondly, to determine pregnancy and cardiac outcomes in women with aortic homograft valves. DESIGN: Historical cohort study. SETTING: Greenlane Hospital, Auckland, New Zealand. POPULATION: Young women (n = 255) who had valve replacements between 1972 and 1992. Seventy-nine women underwent 147 pregnancies. MAIN OUTCOME MEASURES: Pregnancy loss, cardiac complications. RESULTS: Pregnancy loss occurred in 59% of pregnancies with mitral mechanical valves (n = 50) and 7% with mitral bioprosthetic valves (n = 33) (RR 8 x 20, 95% CI 2 x 10-31 x 93). Pregnancy loss rate was 70% in pregnancies treated with warfarin, compared with 25% for those switched from warfarin to heparin (RR 2 x 81, 95% CI 1 x 03-7 x 73). All heparin associated losses occurred in the first trimester, whereas there were four stillbirths with warfarin. Cardiac complications occurred in 10 pregnancies (20%) in the women with mitral mechanical valves and four (13%) with mitral bioprosthetic valves (RR 1 x 55, 95% CI 0 x 53-4 x 52). All four thromboembolic complications with mechanical valves occurred in the 14 women treated with heparin throughout pregnancy. Structural valve deterioration occurred in four pregnancies (10%) with mitral bioprosthetic valves. No cardiac complications or known pregnancy losses occurred with aortic homograft valves (n = 41). CONCLUSION: The high pregnancy loss rate in women with mitral mechanical valves was associated with warfarin throughout pregnancy, whereas the thromboembolic cardiac complications were associated with heparin. Pregnancy outcome was very good in women with bioprosthetic and homograft valves. PMID- 10688510 TI - The outcome of 72 pregnancies in 55 women with cystic fibrosis in the United Kingdom 1977-1996. AB - OBJECTIVE: To identify pregnancies in women with cystic fibrosis and describe obstetric, infant and maternal medical outcomes in relation to the severity of maternal disease. DESIGN: Retrospective study, based on casenotes. SETTING: Eleven cystic fibrosis centres in the United Kingdom. POPULATION: Pregnant women with cystic fibrosis. METHODS: Single observer medical and obstetric casenote review categorising maternal cystic fibrosis (e.g. genotype, pancreatic, hepatic and diabetic status) and pre-pregnant severity (e.g. weight and lung function) and noting fetal outcome and maternal morbidity. MAIN OUTCOME MEASURES: Completed pregnancies and pregnancy losses, fetal outcome and complications, maternal morbidity, such as changes in weight, lung function, pulmonary infections during and after pregnancy. Relation of outcomes to severity of maternal cystic fibrosis. RESULTS: From 72 pregnancies identified, the outcomes were known for 69; there were 48 live births (70%) of which 22 were premature (46%); 14 therapeutic abortions (20%); and 7 miscarriages (10%). There were no stillbirths, neonatal or early maternal deaths. Three major fetal anomalies were seen, but no infant had cystic fibrosis. At the conclusion of our study three pregnancies were still continuing. Prematurity with increased fetal complications and maternal morbidity with infection, declining lung function and poor weight gain were associated with poor pre-partum lung function. CONCLUSION: Pregnancy occurs in women with cystic fibrosis of all degrees of severity. Outcomes for the infant are generally good but are variable for the mother. Predicting outcome on the basis of maternal severity is difficult but lung function appears to be the most significant determining factor. Pregnancy may be normal in women with normal lung function (forced expiratory volume > 80%). However, it may adversely affect mild and moderate lung disease due to cystic fibrosis and should be avoided in pulmonary hypertension, cor pulmonale and when forced expiratory volume < 50% predicted. Ideally, all pregnancies should be planned with prior counselling and monitored by dedicated cystic fibrosis teams, including obstetricians who are experienced in managing high risk pregnancies. PMID- 10688511 TI - Effectiveness of prenatal chromosomal analysis using multicolor fluorescent in situ hybridisation. AB - OBJECTIVE: To evaluate the clinical effectiveness of multicolour fluorescent in situ hybridisation (FISH) analysis in routine prenatal diagnosis. DESIGN: Prospective study. SAMPLE: 3,203 amniotic fluid samples. METHODS: Unique DNA (chromosomes 13 and 21) and alpha satellite centromeric-specific (chromosomes X, Y and 18) probes were used in two mixes to permit the simultaneous analysis of several chromosomes. The performance of multicolour FISH and conventional cytogenetic analysis was compared. RESULTS: Conventional cytogenetic analysis identified 111 chromosomal abnormalities, of which 94 were potentially detectable by the FISH technique and 97 would be typically associated with neonatal phenotypic abnormalities. Multicolour FISH analysis detected 84% (93/111) of all chromosome abnormalities and 99% (93/94) of abnormalities where there was a specific probe. The sensitivity of multicolour FISH analysis was 95% (92/97) for chromosomal abnormalities likely to result in an abnormal postnatal outcome. Multiple ultrasound abnormalities were detected in all five cases of clinically relevant chromosomal abnormalities missed by multicolour FISH. FISH results were available within 48 hours and the sample failure rate was 0 x 1% (3/3,202). CONCLUSION: Multicolour FISH analysis is a sensitive and reliable technique for the rapid prenatal diagnosis of chromosomal abnormalities. Examining only five chromosomes allowed 95% of clinically relevant chromosomal abnormalities to be diagnosed correctly. As routine antenatal screening is targeted at the major autosomal trisomies and sex chromosome aneuploidies, multicolour FISH analysis may potentially replace conventional cytogenetic analysis in routine prenatal diagnosis. PMID- 10688512 TI - Prostaglandin endoperoxide H synthase mRNA expression in the fetal membranes correlates with fetal fibronectin concentration in the cervico-vaginal fluids at term: evidence of enzyme induction before the onset of labour. AB - OBJECTIVE: To determine the relationship of prostaglandin endoperoxide H synthase (PGHS) expression in the gestational tissues and fetal fibronectin in cervico vaginal fluids before the onset of labour at term. DESIGN: Cross-sectional, observational study. SAMPLES: Amnion, chorion laeve and decidua were collected from 24 term pregnant women following elective caesarean section. Samples of cervico-vaginal secretions were obtained from the same women immediately before caesarean section. METHODS: PGHS-1 and PGHS-2 mRNA levels in tissues were determined by specific ribonuclease protection assays. Fetal fibronectin concentrations in the cervico-vaginal fluids were measured by enzyme-linked immunosorbent assay. The abundance of PGHS mRNA was compared between groups of patients with the same mean gestational age but different cervico-vaginal fetal fibronectin levels. Linear regression analysis was used to determine the association between PGHS levels and fetal fibronectin. RESULTS: Two groups of women were identified who had significantly different fetal fibronectin values but the same gestational ages. The group with the higher fetal fibronectin concentrations had significantly higher PGHS- 1 and PGHS-2 mRNA levels in the chorion laeve and higher PGHS-2 mRNA levels in the amnion, than the group with lower fetal fibronectin concentrations. PGHS- 1 and PGHS-2 mRNA levels in the chorion laeve and PGHS-2 mRNA in the amnion showed an overall significant association with fetal fibronectin levels. CONCLUSIONS: High concentrations of fetal fibronectin in cervico-vaginal secretions before the onset of spontaneous labour at term are associated with high levels of PGHS-2 mRNA in the chorion laeve and the amnion and of PGHS- 1 mRNA in the chorion laeve. Increased expression of PGHS in these tissues may therefore be involved in the events leading to term birth. PMID- 10688513 TI - The RUMI manipulator and Koh colpotomiser system for total laparoscopic hysterectomy. AB - The initial experience in 25 patients of using the Koh Colpotomiser System in conjunction with the RUMI Manipulator, a new modified technique for performing total laparoscopic hysterectomy, is presented. Of 25 operations, 23 (92%) were completed successfully. Complications were limited to minor pre-operative haemorrhage in two patients and post-operative bleeding in another. The Koh Colpotomiser System successfully maintained a pneumoperitoneum following colpotomy, giving the operator improved visibility and access to the pelvic organs. This resulted in greater efficiency, while eliminating the difficulties of vaginal access. PMID- 10688514 TI - The relation between tissue kallikrein excretion rate, aldosterone and glomerular filtration rate in human pregnancy. AB - The changes in renal kallikrein synthesis through normal pregnancy and its relation to aldosterone excretion and the glomerular filtration rate was investigated. Overnight urinary kallikrein and aldosterone excretion rates and glomerular filtration rate were measured at 18 weeks, 34 weeks, term and postpartum in normal human pregnancy. Kallikrein excretion was raised at 18 weeks compared with the nonpregnant state (P < 0 x 001) but was significantly reduced at term. The reduction in renal kallikrein was not due to falling aldosterone concentration, which increased in the third trimester, compared with 18 weeks (P = 0 x 002) and remained elevated at term compared with the nonpregnant state (P < 0 x 001). Glomerular filtration rate remained elevated at term despite the reduced kallikrein excretion rate. These data are consistent with the hypothesis that increased aldosterone production is one factor responsible for increased kallikrein synthesis which contributes to elevated glomerular filtration rate in early pregnancy. Other factors clearly inhibit renal kallikrein production at term. In the face of plasma volume expansion associated with increased mineralo corticoid production, the effects of reduced kallikrein synthesis at term on glomerular perfusion and reabsorption of sodium by the distal tubule may predispose to blood pressure elevation in late pregnancy. PMID- 10688515 TI - Good prognosis for psychomotor development in survivors with nonimmune hydrops fetalis. AB - We examined the psychomotor development of 33 of 61 surviving children, from a series of 107 consecutive live-born cases with nonimmune hydrops fetalis. The majority had a normal outcome. Three had a (simultaneous) serious underlying disease (e.g. fetal herpes infection) and had either severe psychomotor retardation or blindness. Two showed clumsiness and were considered to have minor neurological dysfunction. We conclude that survivors, especially those with transient benign intrauterine conditions, such as lymphatic aetiology have no additional risk to their psychomotor development. PMID- 10688516 TI - Folic acid supplements are more effective than increased dietary folate intake in elevating serum folate levels. AB - In 1992, recommendations were disseminated aimed at reducing the incidence of neural tube defects. Women were advised to increase consumption of folic acid supplements and dietary folates during the periconceptional period and a major integrated national campaign was launched to help achieve this. In this study we found that only one-quarter of the women with an uncomplicated obstetric history and 51% with a complicated obstetric history took supplements for the recommended time period. Dietary modification was extremely unusual. Serum analysis demonstrated that intake of folic acid supplements provides a greater elevation in serum folate levels than dietary food intake, suggesting dietary manipulation is an ineffective strategy and that efforts would be better focused on increasing supplement intake at a clinically important time. More effective education strategies are required, and since approximately one-third of pregnancies are unplanned, fortification of foods with folic acid is warranted. PMID- 10688517 TI - Effects of growth hormone treatment in a woman with growth hormone deficiency. PMID- 10688518 TI - Vulvar syringoma showing progesterone receptor positivity. PMID- 10688519 TI - Transvaginal ultrasonography and endometrial histology in peri- and postmenopausal women on hormone replacement therapy. PMID- 10688520 TI - Prophylactic administration of clindamycin 2% vaginal cream to reduce the incidence of spontaneous preterm birth in women with an increased risk: a randomised placebo-controlled double-blind trial. PMID- 10688521 TI - Maternal serum alphafetoprotein screening for open neural tube defects: revised statistical parameters. PMID- 10688522 TI - Review and assessment of selection criteria used when booking pregnant women at different places of birth. PMID- 10688523 TI - Risk factors for rape, physical assault, and posttraumatic stress disorder in women: examination of differential multivariate relationships. AB - The National Women's Study, a 2-year, three-wave longitudinal investigation, employed a national probability sample of 3,006 adult women to: (a) identify separate risk factors for rape and physical assault, and (b) identify separate risk factors associated with post-rape posttraumatic stress disorder (PTSD) and post-physical assault PTSD. This investigation differed from previous studies in that it prospectively examined risk factors at the multivariate, as opposed to univariate level. Overall, past victimization, young age, and a diagnosis of active PTSD increased women's risk of being raped. By contrast, past victimization, minority ethnic status, active depression, and drug use were associated with increased risk of being physically assaulted. Risk factors for PTSD following rape included a history of depression, alcohol abuse, or experienced injury during the rape. However, risk factors for PTSD following physical assault included only a history of depression and lower education. PMID- 10688524 TI - Does anxiety mitigate the behavioral expression of severe conduct disorder in delinquent youths? AB - The purpose of this article is to examine the purported attenuating effects of comorbid anxiety on conduct disturbance in a sample of youths exhibiting severe Conduct Disorder (CD). Further, we examined the differential expression of CD and comorbid anxiety in male and female youths. Seventy-nine incarcerated youths between the ages of 12 and 19 were interviewed using the Diagnostic Interview Schedule for Children. Youths were identified who exhibited CD and CD plus an anxiety disorder. In contrast to previous findings with younger, less seriously disturbed male subjects, no overall differences were found between CD anxious and CD nonanxious youths in terms of age of first offense and overall number and severity of delinquent acts. Moreover, no differences were found between males and females, and gender did not moderate the effects of comorbidity anxiety on outcome measures. Findings suggest purported mitigating effects of anxiety on conduct disturbance may be attenuated in severe forms of CD and support the notion that comorbidity across internalizing and externalizing domains of child and adolescent psychopathology may differentially impact clinical presentation of disordered behavior depending on the severity of externalizing behavioral disturbance. PMID- 10688525 TI - The efficacy of habituation in decreasing subjective distress among high anxiety sensitive college students. AB - While there is mounting evidence that the concept of anxiety sensitivity (AS) is linked to the expression of anxiety (specifically, panic), there has been little research comparing the efficacy of interoceptive exposure alone with interoceptive exposure coupled with cognitive restructuring among high AS participants. The present investigation addressed this issue in a sample of high anxiety-sensitive college students (scores above 29 on the Anxiety Sensitivity Index). Participants were randomly assigned to receive either five consecutive trials of voluntary hyperventilation or five consecutive trials of hyperventilation with cognitive restructuring instructions. It was expected that while repeated hyperventilation would be associated with a significant reduction in self-reported anxiety, catastrophic cognitions, and somatic sensations across trials, the greatest reduction in symptoms would occur with the addition of cognitive restructuring. These predictions were partially supported. As expected, high AS participants evidenced significant decreases in anxiety symptoms when habituation was accompanied by cognitive restructuring. Contrary to predictions, however, interoceptive exposure alone was not effective in reducing anxious symptoms. These results suggest that brief habituation alone may not be an effective strategy for high AS participants and are discussed as providing further support for a cognitive model of anxiety. PMID- 10688526 TI - The effects of time pressure on arithmetic performance. AB - It has recently been demonstrated that highly math-anxious individuals may be less proficient on arithmetic tasks, particularly those that involve complex problems. The processing efficiency theory postulates that in highly anxious individuals, working memory resources are consumed by "worry," thereby leaving fewer resources available for task completion. Although there is some empirical support for this theory, the precise nature of this worry has yet to be identified. We tested the hypothesis that time pressure may be one component contributing to worry, and subsequent performance deficits characteristic of high math-anxious individuals. Thirty participants completed arithmetic problems of varying complexity in both a timed and untimed condition. Although the timing manipulation negatively affected arithmetic performance in both high and low anxious participants, anxiety groups were not differentially affected. Researchers may therefore have to look to other variables to explain the nature of worrisome thoughts that are theorized to disrupt the performance of anxious individuals. PMID- 10688527 TI - Levels of anxiety sensitivity in relation to repressive and self-deceptive coping styles. AB - Levels of anxiety sensitivity (AS) were investigated in relation to self deception and repression in 296 university students. It was hypothesized that a low level of AS, rather than constituting "normal" functioning, would be associated with more general response biases. Scores on the Anxiety Sensitivity Index (ASI; Peterson & Reiss, 1987, 1992) were negatively and significantly correlated with measures of self-deception and denial. Individuals with low AS were significantly more likely to meet an operational definition of repression (i.e., low anxiety and high defensiveness), compared to mid AS and high AS groups. When confronted with a hypothetical health problem, individuals with low AS were less likely to choose a task-oriented response and more likely to choose denial and self-deceptive responses, compared to the other groups. These findings support the observations of Shostak and Peterson (1990) that low AS represents an extreme group that may not be indicative of normal functioning. PMID- 10688528 TI - The power of suggestion: comment on EMDR and mesmerism: a comparative historical analysis. AB - This response to McNally challenges the notion that scientific controversy should be waged with smear tactics. McNally's anti-EMDR conclusions are contested as premature and based on red herrings, selective neglect of the literature, and erroneous application of scientific principles. The importance of treatment fidelity is highlighted as a way of distinguishing between EMDR studies of widely varying quality. PMID- 10688529 TI - Altered electrophoretic migration of polycyclic aromatic hydrocarbon and styrene oxide adducts at adenine N(6) correlates with adduct-induced structural disorder. AB - Site-specific bay region benzo[a]pyrene (7R,8S,9R,10S)-N(6)-[10-(7,8, 9,10 tetrahydro-7,8,9-trihydroxybenzo[a]pyrenyl)]-2'-deoxyadeno syl, (7S,8R,9S,10R) N(6)-[10-(7,8,9,10-tetrahydro-7,8, 9-trihydroxybenzo[a]pyrenyl)]-2' deoxyadenosyl, (7S,8R,9R, 10S)-N(6)-[10-(7,8,9,10-tetrahydro-7,8, 9 trihydroxybenzo[a]pyrenyl)]-2'-deoxyadenosyl, and (7R,8S,9S, 10R)-N(6)-[10 (7,8,9,10-tetrahydro-7,8, 9-trihydroxybenzo[a]pyrenyl)]-2'-deoxyadenosyl adducts, bay region benz[a]anthracene (1R,2S,3R,4S)-N(6)-[1-(1,2,3,4-tetrahydro-2,3, 4 trihydroxybenz[a]anthracenyl)]-2'-deoxyadenosyl and (1S,2R,3S, 4R)-N(6)-[1 (1,2,3,4-tetrahydro-2,3, 4-trihydroxybenz[a]anthracenyl)]-2'-deoxyadenosyl adducts, non-bay region benz[a]anthracenyl (8S,9R,10S,11R)-N(6)-[11-(8,9,10, 11 tetrahydro-8,9,10-trihydroxybenz[a]anthracenyl)]-2'-de oxyadenosyl and (8R,9S,10R,11S)-N(6)-[11-(8,9,10,11-tetrahydro-8,9, 10 trihydroxybenz[a]anthracenyl)]-2'-deoxyadenosyl adducts, and the R- and S-adducts of styrene oxide were located in the ras61 oligodeoxynucleotide and examined with respect to electrophoretic mobility. The results were compared to NMR structural data, and to site-specific mutagenesis data and in vitro DNA replication assays for the same adducts. There was a correlation between adducts having lower electrophoretic mobility and greater disorder at the adduct site as monitored by NMR. The disorder combined with the lower electrophoretic mobilities suggested that these adducts induced flexible hinge joints in the DNA rather than static bending. Usually, these were adenine N(6) adducts having S-stereochemistry at the benzylic carbon. The results also revealed a possible role for the bay region ring in stabilizing adenyl N(6) benz[a]anthracene adducts with respect to hinging at the adduct site. On the other hand, there was not a simple relationship between altered electrophoretic mobility and mutagenesis or DNA replication. PMID- 10688530 TI - In vitro DNA deamination by alpha-nitrosaminoaldehydes determined by GC/MS-SIM quantitation. AB - The deamination of DNA bases by three alpha-nitrosaminoaldehydes, butylethanalnitrosamine, methylethanalnitrosamine, and N-nitroso-2 hydroxymorpholine (NHMOR), the direct metabolite of potent animal carcinogen N nitrosodiethanolamine, was demonstrated by a set of in vitro experiments. The deamination of guanine, adenine, and cytosine bases in nucleotides, oligonucleotides, and calf thymus DNA gave xanthine, hypoxanthine, and uracil, respectively. The order of relative reactivities of the bases was as listed above. Deamination of cytosine to uracil was detected by the reaction of (32)P labeled oligonucleotide ([5'-(32)P]CGAT) followed by enzymatic hydrolysis. Quantitative analysis of deamination of guanine and adenine in calf thymus DNA was performed by a gas chromatography/mass spectrometry-selected ion monitoring method. Both the extent and the rate of the deamination reactions which occur by transnitrosation from the alpha-nitrosaminoaldehyde to the base were determined for formation of xanthine and hypoxanthine. The deamination of guanine by NHMOR remained significant at low substrate levels. PMID- 10688531 TI - Isocyanate-specific hemoglobin adduct in rats exposed to 4, 4'-methylenediphenyl diisocyanate. AB - 4,4'-Methylenediphenyl diisocyanate (MDI) is the most important of the isocyanates used as intermediates in the chemical industry. Among the main types of damage after exposure to low levels of MDI are lung sensitization and asthma. Protein adducts of MDI might be involved in the etiology of sensitization reactions. It is therefore necessary to have sensitive and specific methods for monitoring the isocyanate exposure of workers. To date, urine metabolites or protein adducts have been used as biomarkers in workers exposed to MDI. However, with these methods it is not possible to determine if the biomarkers result from exposure to MDI or to the parent aromatic amine 4,4'-methylenedianiline (MDA). This work presents a procedure for quantitating isocyanate-specific hemoglobin adducts. Blood proteins are used as markers of exposure and possibly as markers of dose size for the modifications of macromolecules in the target organs where the disease develops. For the quantitation of hemoglobin adducts, N(1)-[4-(4 isocyanatobenzyl)phenyl]acetamide (AcMDI) was reacted with the tripeptide valyl glycyl-glycine and with valine yielding N-[4-(4 acetylaminobenzyl)phenyl]carbamoyl]valyl-glycyl-glycine and N-[4-[4 (acetylaminobenzyl)phenyl]carbamoyl]valine, respectively. N-[4-[4-(Acetylamino 3,5-dideuteriobenzyl)-2, 6-dideuteriophenyl]carbamoyl]valine was synthesized from valine, as was N(1)-[4-(4-isocyanato-3,5-dideuteriobenzyl)-2, 6 dideuteriophenyl]acetamide, for use as an internal standard. These adducts were cleaved in 2 M HCl to yield the corresponding hydantoins, 3-[4-(4 aminobenzyl)phenyl]-5-isopropyl-1, 3-imidazoline-2,4-dione (MDA-Val-Hyd) and 3-[4 (4-amino-3, 5-dideuteriobenzyl)-2,6-dideuteriophenyl]-5-isopropyl-1, 3 imidazoline-2,4-dione, respectively. In globin of rats exposed to MDI, MDA-Val Hyd could be found in a dose-dependent manner. The adduct was identified by HPLC/MS/MS and quantified by GC/MS after derivatization with heptafluorobutyric anhydride. The amount of MDA-Val-Hyd found after acid hydrolysis of globin at 100 degrees C is about 12 times larger than the sum of N-acetyl-4, 4' methylenedianiline (AcMDA) and MDA obtained from mild base hydrolysis of hemoglobin. The MDA-Val-Hyd is an isocyanate-specific adduct. MDA and AcMDA released after mild base hydrolyses result most likely from a sulfinamide adduct which is a typical adduct of arylamines. According to these results, higher amounts of isocyanate adducts than arylamine adducts should be expected in workers exposed to isocyanates. PMID- 10688532 TI - Synthesis of oligonucleotides containing the alkali-labile pyrimidopurinone adduct, M(1)G. AB - An improved method for the synthesis of oligodeoxyribonucleotides containing the endogenous adduct, pyrimido[1,2-a]purin-10(3H)-one (M(1)G), is reported. The key features of the methodology include improved synthesis of the deoxynucleoside of M(1)G by transribosylation with deoxycytidine catalyzed by nucleoside 2' deoxyribosyltransferase and the use of commercially available 4-tert butylphenoxyacetyl protecting groups for normal nucleotides. Facile deprotection and removal of the M(1)G-containing oligomers from the solid support were achieved by treatment with a solution of potassium carbonate in methanol. NMR studies were performed to determine the stability of the oligonucleotides at different pHs. PMID- 10688533 TI - Sulfinamide formation following peroxidatic metabolism of N-acetylbenzidine. AB - Arylamine-hemoglobin conjugates identified as sulfinamides are considered dosimeters for the bioavailability of metabolically formed N-oxidation products. This report considers peroxidation as an alternative pathway for aromatic amine metabolism and examines horseradish peroxidase metabolism of N-acetylbenzidine (ABZ) in the presence of glutathione. When 0.06 mM [(3)H]ABZ was incubated with 1 mM glutathione, a decrease in the total extent of metabolism was observed along with detection of a new metabolite (ABZ-SG), representing 12% of the total radioactivity. Optimum ABZ-SG formation occurred at 0.3 mM glutathione with higher concentrations (10 mM) being inhibitory. In the absence of glutathione, a molar ratio of H(2)O(2) to ABZ of 1:1 resulted in complete metabolism of ABZ. This ratio increased to >2:1 in the presence of 0.3 mM glutathione. N-Oxidation products of ABZ metabolism, such as N'-hydroxy-N-acetylbenzidine, were not detected using a variety of incubation conditions. ABZ-SG was sensitive to gamma glutamyltranspeptidase, and completely hydrolyzed by 0.1 N HC1 or 0.1 N NaOH in 10 min at room temperature. ABZ-SG was identified by mass spectrometry and NMR to be N'-(glutathion-S-yl)-N-acetylbenzidine S-oxide. ABZ-SG formation, but not total ABZ metabolism, was prevented by 0.3 mM NaN(3), 50 mM DMPO, 1.0 mM thiourea, and 1.0 mM histidine. Cyanide (50 mM) and ascorbic acid (0.1 mM) completely inhibited ABZ metabolism. The lack of effect of 50 mM mannitol and 2 microgram of superoxide dismutase suggests that neither hydroxyl radical nor superoxide is involved in the reaction. Studies also indicated that molecular oxygen is not a source of the sulfinamide oxygen. Formation of an ABZ sulfinamide conjugate with hemoglobin was demonstrated. The proposed mechanism for sulfinamide formation, involving two consecutive one-electron oxidations with subsequent rearrangement to a sulfur-stabilized nitrenium ion, suggests that oxygen may be derived from water. The results demonstrate that while arylamine hemoglobin conjugates serve as useful biomarkers of exposure, their mechanism of formation may be complex, perhaps involving peroxidation as in the case of N' (glutathion-S-yl)-N-acetylbenzidine S-oxide. PMID- 10688534 TI - The sensitivity of carboxyl-terminal methionines in calmodulin isoforms to oxidation by H(2)O(2) modulates the ability to activate the plasma membrane Ca ATPase. AB - The oxidative modification of methionines within the primary sequence of calmodulin (CaM) results in an inability to activate the PM-Ca-ATPase fully, and may contribute to alterations in calcium homeostasis under conditions of oxidative stress. To identify differences in the sensitivities of CaM isoforms to oxidative modification, we have compared the function and patterns of oxidative modification resulting from the exposure of CaM isolated from bovine testes and wheat germ to H(2)O(2). In comparison to CaM isolated from wheat germ, vertebrate CaM is functionally resistant to oxidant-induced loss of function. The decreased functional sensitivity of vertebrate CaM correlates with a 75 +/- 3% reduction in the rate of oxidative modification of a methionine near the carboxyl terminus (i.e., Met(144) or Met(145)). The extent of oxidative modification to other methionines in these CaM isoforms is similar. These results suggest that the sensitivity of Met(144) or Met(145) to oxidation modulates the ability of CaM to activate the PM-Ca-ATPase. Consistent with this interpretation, a CaM mutant in which glutamines were substituted for Met(144) and Met(145) fully activates the PM-Ca-ATPase irrespective of the oxidative modification of the other seven methionines to their corresponding methionine sulfoxides. The extent of oxidative modification to individual methionines in vertebrate CaM by H(2)O(2) correlates with the time-averaged surface accessibility of individual sulfurs calculated from molecular dynamics simulations. Thus, the sensitivity of individual methionines to oxidative modification is directly related to the solvent accessibility. These results indicate that sequence differences between vertebrate and plant CaM alter the sensitivity of methionines near the carboxyl terminus to oxidative modification because of alterations in their solvent accessibility. We suggest that these sequence differences between CaM isoforms have a regulatory role in modulating the functional sensitivity of CaM to conditions of oxidative stress. PMID- 10688535 TI - Stress- and growth-related gene expression are independent of chemical-induced prostaglandin E(2) synthesis in renal epithelial cells. AB - Cellular stress can initiate prostaglandin (PG) biosynthesis which, through changes in gene expression, can modulate cellular functions, including cell growth. PGA(2), a metabolite of PGE(2), induces the expression of stress response genes, including gadd153 and hsp70, in HeLa cells and human diploid fibroblasts. PGs, gadd153, and hsp70 expression are also influenced by the cellular redox status. Polyphenolic glutathione conjugates retain the ability to redox cycle, with the concomitant generation of reactive oxygen species. One such conjugate, 2,3,5-tris(glutathion-S-yl)hydroquinone (TGHQ), is a potent nephrotoxic and nephrocarcinogenic metabolite of the nephrocarcinogen, hydroquinone. We therefore investigated the effects of TGHQ on PGE(2) synthesis and gene expression in a renal proximal tubular epithelial cell line (LLC-PK(1)). TGHQ (200 microM, 2 h) increases PGE(2) synthesis (2-3-fold) in LLC-PK(1) cells with only minor (5%) reductions in cell viability. This response is toxicant-specific, since another proximal tubular toxicant, S-(1, 2-dichlorovinyl)-L-cysteine (DCVC), stimulates PGE(2) synthesis only after massive (68%) reductions in cell viability. Consistent with the ability of TGHQ to generate an oxidative stress, both deferoxamine mesylate and catalase protect LLC-PK(1) cells from TGHQ-mediated cytotoxicity. Only catalase, however, completely blocks TGHQ-mediated PGE(2) synthesis, implying a major role for hydrogen peroxide in this response. TGHQ induces the early (60 min) expression of gadd153 and hsp70. However, while inhibition of cyclooxygenase with aspirin prevents TGHQ-induced PGE(2) synthesis, it does not affect TGHQ-mediated induction of gadd153 or hsp70 expression. In contrast, a stable PGE(2) analogue, 11-deoxy-16, 16-dimethyl-PGE(2) (DDM-PGE(2)), which protects LLC-PK(1) cells against TGHQ-mediated cytotoxicity, modestly elevates the levels of gadd153 and hsp70 expression. In addition, catalase and, to a lesser extent, deferoxamine mesylate block TGHQ-induced gene expression. Therefore, although TGHQ-induced generation of reactive oxygen species is required for PGE(2) synthesis and stress gene expression, acute TGHQ-mediated increases in gadd153 and hsp70 mRNA levels are independent of PGE(2) synthesis. PMID- 10688536 TI - Interleukin-8 levels in human lung epithelial cells are increased in response to coal fly ash and vary with the bioavailability of iron, as a function of particle size and source of coal. AB - Particulate air pollution contains iron, and some of the pathological effects after inhalation may be due to radical species produced by iron-catalyzed reactions. We tested the hypothesis that iron present in coal fly ash (CFA) could induce the expression and synthesis of the inflammatory cytokine interleukin-8 (IL-8). CFA, containing as much as 14% iron, was used as a model combustion source particle. Three coal types were used to generate three size fractions enriched in particles [submicron (<1 micrometer), fine (<2.5 micrometer), or coarse (2.5-10 micrometer]), as well as the fraction of >10 micrometer. Treatment of human lung epithelial (A549) cells for 4 h with CFA from Utah enriched in <1 micrometer particles (20 microgram/cm(2)) resulted in a 2.6-fold increase in mRNA levels for IL-8. IL-8 levels were increased in the medium by as much as 8-fold when cells were treated with the fraction enriched in the smallest size Utah CFA for 24 h. IL-8 production was completely inhibited when the CFA was pretreated with the metal chelator desferrioxamine B, suggesting that a transition metal was responsible for the induction, probably iron. Treatment with a soluble form of iron, ferric ammonium citrate (FAC), mimicked the IL-8 level increase observed with CFA. There was a direct relationship, above a threshold level of bioavailable iron, between the levels of IL-8 and bioavailable iron in A549 cells treated with CFA or FAC. Further, the relationship between IL-8 and bioavailable iron for CFA was indistinguishable from that for FAC. These results strongly suggest that iron can induce IL-8 in A549 cells and that iron was the likely component of CFA that induced IL-8. CFA-induced IL-8 production was inhibited by tetramethylthiourea or dimethyl sulfoxide, suggesting that radical species were involved in the induction. These results demonstrate that iron present in CFA may be responsible for production and release of inflammatory mediators by the lung epithelium through generation of radical species and suggest that iron may contribute to the exacerbation of respiratory problems by particulate air pollution. PMID- 10688537 TI - Rat liver cytosol catalyzes a reaction involving activated N-nitrosodimethylamine and a carbohydrate from the pentose phosphate pathway. AB - N-Nitrosodimethylamine is a liver toxin and mutagen following activation by cytochrome P450. The role of the cytosol in N-nitrosodimethylamine metabolism is not well understood. The effect of cytosol on N-nitrosodimethylamine metabolism was investigated using microsomes and cytosol from rat liver in in vitro reactions with N-nitrosodimethylamine and an NADPH generating system. Studies in which [(14)C]-N-nitrosodimethylamine and calf thymus DNA were used indicated that the addition of cytosol to the microsomal reaction mixture resulted in >200% enhancement of the radioactivity associated with DNA after the DNA was isolated from the reaction mixture by phenol extraction followed by ethanol precipitation. This stimulatory effect was associated with a cytosolic protein and was found to be dependent on both the microsomes and the carbohydrate used in the glucose-6 phosphate dehydrogenase system for the generation of NADPH. The carbohydrate requirement was found to be specific for intermediates of the pentose phosphate pathway, and maximum stimulation occurred with ribulose 5-phosphate. Most of the counts from [(14)C]-N-nitrosodimethylamine which were isolated with DNA after the addition of cytosol to reaction mixtures were not covalently bound to the DNA. HPLC analysis identified four radiolabeled metabolites derived from [(14)C]-N nitrosodimethylamine following the in vitro incubations. One of the four products was formed only when both cytosol and ribulose 5-phosphate were added to the enzymatic incubations. This product also formed from [(14)C]-alpha-acetoxy nitrosodimethylamine in the absence of microsomes, only when cytosol and ribulose 5-phosphate were added to the reaction mixtures. Thus, these data demonstrate that an enzyme in the cytosol catalyzes a reaction involving a metabolite of N nitrosodimethylamine (which is formed following cytochrome P450-mediated activation) and a carbohydrate related to the pentose phosphate pathway. A similar reaction also occurs with N-diethylnitrosamine but not with N dipropylnitrosamine or N-dibutylnitrosamine. PMID- 10688538 TI - Advances in managing chronic disease. Research, performance measurement, and quality improvement are key. PMID- 10688539 TI - Patients as partners in managing chronic disease. Partnership is a prerequisite for effective and efficient health care. PMID- 10688540 TI - Depression management clinics in general practice? Some aspects lend themselves to the mini-clinic approach. PMID- 10688541 TI - Building evidence on chronic disease in old age. Standardised assessments and databases offer one way of building the evidence. PMID- 10688542 TI - Disease management: has it a future? It has a compelling logic, but needs to be tested in practice. PMID- 10688543 TI - British tobacco company denies "orchestrating smuggling". PMID- 10688544 TI - Paediatricians "close to despair" over child protection laws PMID- 10688545 TI - UK home secretary faces new problems over pinochet PMID- 10688546 TI - Screening changes recommended for osteoporosis and diabetes PMID- 10688547 TI - In brief PMID- 10688548 TI - Albumin industry launches global promotion. PMID- 10688549 TI - Britain's health league table attacked. PMID- 10688550 TI - Hospital infection rates in England out of control. PMID- 10688551 TI - Donors and relatives must place no conditions on organ use. PMID- 10688552 TI - Gynaecologist cleared in hysterectomy case. PMID- 10688553 TI - Spain's home healthcare programme goes nationwide. PMID- 10688556 TI - Cirrhosis may be amenable to telomerase treatment PMID- 10688554 TI - British Columbia blocks tobacco move. PMID- 10688557 TI - Painkillers "may need to be sex specific". PMID- 10688558 TI - Systematic reviews and meta-analyses on treatment of asthma: critical evaluation. AB - OBJECTIVE: To evaluate the clinical, methodological, and reporting aspects of systematic reviews and meta-analyses on the treatment of asthma and to compare those published by the Cochrane Collaboration with those published in paper based journals. DESIGN: Analysis of studies identified from Medline, CINAHL, HealthSTAR, EMBASE, Cochrane Library, personal collections, and reference lists. STUDIES: Articles describing a systematic review or a meta-analysis of the treatment of asthma that were published as a full report, in any language or format, in a peer reviewed journal or the Cochrane Library. MAIN OUTCOME MEASURES: General characteristics of studies reviewed and methodological characteristics (sources of articles; language restrictions; format, design, and publication status of studies included; type of data synthesis; and methodological quality). RESULTS: 50 systematic reviews and meta-analyses were included. More than half were published in the past two years. Twelve reviews were published in the Cochrane Library and 38 were published in 22 peer reviewed journals. Forced expiratory volume in one second was the most frequently used outcome, but few reviews evaluated the effect of treatment on costs or patient preferences. Forty reviews were judged to have serious or extensive flaws. All six reviews associated with industry were in this group. Seven of the 10 most rigorous reviews were published in the Cochrane Library. CONCLUSIONS: Most reviews published in peer reviewed journals or funded by industry have serious methodological flaws that limit their value to guide decisions. Cochrane reviews are more rigorous and better reported than those published in peer reviewed journals. PMID- 10688559 TI - Qualitative study of interpretation of reassurance among patients attending rheumatology clinics: "just a touch of arthritis, doctor?". AB - OBJECTIVES: To examine commonly used methods of reassurance by clinicians and explore their effect on patients. DESIGN: Qualitative study of tape recordings of in-depth, semistructured interviews with patients before and after consultation and of their consultations with doctors. SETTING: NHS specialist rheumatology clinics in two large British cities. PARTICIPANTS: 35 patients selected by consultant rheumatologists from general practitioner referral letters (28 women, 7 men; 24 with inflammatory arthropathies, 11 other rheumatological complaints). MAIN OUTCOME MEASURES: Patients' perceptions of reassurance. RESULTS: Reassurance was an important part of consultations, whether the diagnosis was clear or uncertain. Clinicians tried to reduce anxiety by emphasising the mildness, early stage, or non-seriousness of the disorder and the likelihood that patients would recover. Patients interpreted reassurance in the context of their own views and perceptions. Doctors' emphasis on the mildness or earliness of the condition raised the spectre of future pain and disability rather than providing reassurance. Patients who felt that their problems were properly acknowledged felt more reassured. CONCLUSIONS: Typical patterns of reassurance were not successful because of the differences in perspective of patients and doctors. A key to successful reassurance seemed to be the doctor's ability to acknowledge patients' perspectives of their difficulties. PMID- 10688560 TI - Open access follow up for inflammatory bowel disease: pragmatic randomised trial and cost effectiveness study. AB - OBJECTIVE: To evaluate whether follow up of patients with inflammatory bowel disease is better through open access than by routine booked appointments. DESIGN: Pragmatic randomised controlled trial. SETTING: Two district general hospitals in Swansea and Neath, Wales. PARTICIPANTS: 180 adults (78 with Crohn's disease, 77 ulcerative or indeterminate colitis, 25 ulcerative or idiopathic proctitis) recruited from outpatient clinics during October 1995 to November 1996. INTERVENTION: Open access follow up according to patient need. MAIN OUTCOME MEASURES: Generic (SF-36) and disease specific (UK inflammatory bowel disease questionnaire UKIBDQ) quality of life, number of primary and secondary care contacts, total resource use, and views of patients and general practitioners. RESULTS: There were no differences in generic or disease specific quality of life. Open access patients had fewer day visits (0.21 v 0. 42, P<0.05) and fewer outpatient visits ( 4.12 v 4.64, P<0.01), but some patients had difficulty obtaining an urgent appointment. There were no significant differences in specific investigations undertaken, inpatient days, general practitioner surgery or home visits, drugs prescribed, or total patient borne costs. Mean total cost in secondary care was lower for open access patients (P<0.05), but when primary care and patient borne costs were added there were no significant differences in total costs to the NHS or to society. General practitioners and patients preferred open access. CONCLUSIONS: Open access follow up delivers the same quality of care as routine outpatient care and is preferred by patients and general practitioners. It uses fewer resources in secondary care but total resource use is similar. Better methods of ensuring urgent access to outpatient clinics are needed. PMID- 10688561 TI - Five year follow up of a randomised controlled trial of a stroke rehabilitation unit. PMID- 10688562 TI - Machiavelli on clinical management PMID- 10688563 TI - Randomised trial of monitoring, feedback, and management of care by telephone to improve treatment of depression in primary care. AB - OBJECTIVE: To test the effectiveness of two programmes to improve the treatment of acute depression in primary care. DESIGN: Randomised trial. SETTING: Primary care clinics in Seattle. PATIENTS: 613 patients starting antidepressant treatment. INTERVENTION: Patients were randomly assigned to continued usual care or one of two interventions: feedback only and feedback plus care management. Feedback only comprised feedback and algorithm based recommendations to doctors on the basis of data from computerised records of pharmacy and visits. Feedback plus care management included systematic follow up by telephone, sophisticated treatment recommendations, and practice support by a care manager. MAIN OUTCOME MEASURES: Blinded interviews by telephone 3 and 6 months after the initial prescription included a 20 item depression scale from the Hopkins symptom checklist and the structured clinical interview for the current DSM-IV depression module. Visits, antidepressant prescriptions, and overall use of health care were assessed from computerised records. RESULTS: Compared with usual care, feedback only had no significant effect on treatment received or patient outcomes. Patients receiving feedback plus care management had a higher probability of both receiving at least moderate doses of antidepressants (odds ratio 1.99, 95% confidence interval 1.23 to 3.22) and a 50% improvement in depression scores on the symptom checklist (2.22, 1.31 to 3.75), lower mean depression scores on the symptom checklist at follow up, and a lower probability of major depression at follow up (0.46, 0.24 to 0.86). The incremental cost of feedback plus care management was about $80 ( pound50) per patient. CONCLUSIONS: Monitoring and feedback to doctors yielded no significant benefits for patients in primary care starting antidepressant treatment. A programme of systematic follow up and care management by telephone, however, significantly improved outcomes at modest cost. PMID- 10688564 TI - Management of chronic uveitis. PMID- 10688565 TI - ABC of heart failure. Acute and chronic management strategies. PMID- 10688566 TI - Disease management in the American market. PMID- 10688567 TI - Commercial partnerships in chronic disease management: proceeding with caution. PMID- 10688570 TI - Fascinating rhythm PMID- 10688568 TI - The role of patient care teams in chronic disease management. PMID- 10688569 TI - Management of chronic disease by practitioners and patients: are we teaching the wrong things? PMID- 10688571 TI - Management of hypertension. Ideal body weight is not realistic goal for lifestyle intervention. PMID- 10688572 TI - Greenwich asthma study. Study's conclusions are premature. PMID- 10688573 TI - beta Blockade after myocardial infarction. Beta blockers have key role in reducing morbidity and mortality after infarction. PMID- 10688574 TI - Health professionals do not understand mathematical models. PMID- 10688575 TI - Inequalities in health continue to grow despite government's pledges. PMID- 10688577 TI - PM will address GPs' conference PMID- 10688576 TI - Obituaries PMID- 10688578 TI - Incontinence PMID- 10688579 TI - Schizophrenia: concepts and clinical management PMID- 10688580 TI - Handbook for mortals: guidance for people facing serious illness PMID- 10688581 TI - Epilepsy: problem solving in clinical practice PMID- 10688582 TI - Let the children do the talking PMID- 10688583 TI - Disease management PMID- 10688584 TI - Facing the challenges of long term care PMID- 10688585 TI - And how is sir today? PMID- 10688587 TI - Systematic reviews and meta-analyses are often flawed PMID- 10688586 TI - Management and preferences PMID- 10688589 TI - Open access follow up can work in chronic bowel disease PMID- 10688588 TI - Doctors' reassurances are often ineffective PMID- 10688590 TI - Stroke units seem to have a long term impact on patients' outcomes PMID- 10688591 TI - Monitoring, feedback, and care management improve treatment of depression PMID- 10688592 TI - Doctors and patients are taught the wrong skills for managing chronic disease PMID- 10688593 TI - Histamine H(3)-receptors: a new frontier in myocardial ischemia. AB - In protracted myocardial ischemia, sympathetic nerve endings undergo ATP depletion, hypoxia and pH(i) reduction. Consequently, norepinephrine (NE) accumulates in the axoplasm, because it is no longer stored in synaptic vesicles, and intraneuronal Na(+) concentration increases, as the Na(+)/H(+) exchanger (NHE) is activated. This forces the reversal of the Na(+)- and Cl(-)-dependent NE transporter, triggering a massive carrier-mediated release of NE and thus, arrhythmias. Indeed, NE overflow in myocardial ischemia directly correlates with the severity of arrhythmias. Histamine H(3)-receptors (H(3)R) have been identified as inhibitory heteroreceptors in adrenergic nerve endings of the heart. In addition to inhibiting NE exocytosis from sympathetic nerve endings, selective H(3)R agonists attenuate carrier-mediated release of NE in both animal and human models of protracted myocardial ischemia. Whereas H(3)R-mediated attenuation of exocytotic NE release involves an inhibition of N-type Ca(2+) channels, H(3)R-mediated reduction of carrier-mediated NE release is associated with diminished NHE activity. In addition to inhibiting NE release, H(3)R stimulation significantly attenuates the incidence and duration of ventricular fibrillation. Although other presynaptic receptors also modulate NE release from sympathetic nerve endings, H(3)R stimulation reduces both exocytotic and carrier mediated NE release, whereas alpha(2)-adrenoceptor agonists attenuate NE exocytosis but enhance carrier-mediated NE release. Furthermore, unlike adenosine A(1)-receptors, whose activation reduces both exocytotic and carrier-mediated NE release, H(3)R stimulation is devoid of negative chronotropic and dromotropic effects (i.e., sinoatrial and atrioventricular nodal functions are unaffected). Because excess NE release can trigger severe arrhythmias and sudden cardiac death, negative modulation of NE release by H(3)R agonists may offer a novel therapeutic approach to myocardial ischemia. PMID- 10688594 TI - Anti-phencyclidine monoclonal antibodies provide long-term reductions in brain phencyclidine concentrations during chronic phencyclidine administration in rats. AB - These studies examined the hypothesis that a single large dose of monoclonal anti phencyclidine (PCP) antibody could provide long-term reductions in brain PCP concentrations despite continuous PCP administration. PCP (18 mg/kg/day, s.c.) was infused to steady-state (24 h) and then a mole-equivalent dose of a short acting anti-PCP antigen-binding fragment (Fab) or a long-acting anti-PCP IgG was administered i.v. The PCP infusion continued for up to 27 days, even though the binding capacity of the single dose of antibody used should have been saturated within the first day. At selected time points after antibody administration, brain, testis, and serum PCP concentrations were measured. Serum PCP concentrations rapidly increased approximately 100- and 300-fold after Fab or IgG administration, respectively. Based on the antibody-bound PCP concentrations in serum, the functional elimination half-life (t(1/2lambdaZ)) values for PCP-Fab and PCP-IgG complexes were 9.4 h and 15.4 days, respectively. Fab and IgG administration produced a complete removal of PCP from the brain within 15 min. Although brain PCP concentrations were significantly decreased for only 4 h in Fab-treated animals, IgG administration resulted in significant decreases in brain PCP concentrations lasting for at least 27 days. In contrast, testis PCP concentrations were not substantially affected by antibody administration, suggesting that redistribution of PCP from the testis is too slow to benefit from a limited dose of antibody. These results indicate that anti-PCP IgG can preferentially protect the brain for approximately 4 weeks after IgG administration, even when the antibody binding capacity should have been saturated with continuously administered PCP. PMID- 10688595 TI - Targeted antioxidant properties of N-[(tetramethyl-3-pyrroline-3 carboxamido)propyl]phthalimide and its nitroxide metabolite in preventing postischemic myocardial injury. AB - We investigated the cardioprotective efficacy of a new compound based on 2,2,5,5 tetramethyl-3-pyrroline-3-carboxamide (TPC-NH). Biochemical studies using electron paramagnetic resonance (EPR) spectroscopy suggest that TPC-NH is a scavenger of reactive oxygen species. In vitro cellular studies show that TPC-NH protects isolated cardiomyocytes against oxidative damage caused by superoxide radicals. Ex vivo EPR studies on the isolated rat heart indicate that the TPC-NH is metabolically oxidized to the nitroxide form. Studies were also performed in the isolated rat heart model to measure the efficacy of TPC-NH and its metabolites in preventing postischemic reperfusion injury. Serial measurements of contractile function were performed on hearts subjected to ischemia-reperfusion. Hearts were either untreated or treated with 50 microM TPC-NH or with its metabolites for 1 min before ischemia and during the first 5 min of reflow. TPC NH showed marked protection with a more than 3-fold increased recovery of contractile function compared with control hearts, whereas its oxidative metabolites exhibited significant but lower protection. Thus, TPC-NH and, to a lesser extent, its oxidation metabolites exhibit potent membrane-targeted antioxidant action and exert marked protection against myocardial injury in the postischemic heart. PMID- 10688596 TI - Disposition of dodecanedioic acid in humans. AB - The disposition of dodecanedioic acid (C12) was investigated in six overnight fasting healthy male volunteers, who received a 165-min i. v. infusion of 42.45 mmol of C12 added to 150 microCi of [1-12-(14)C]C12. Blood samples were collected up to 360 min after the start of infusion, and concentration of serum labeled C12 was determined. Expired radioactivity (microCi/min) was measured up to 600 min and at 24 h. The 24-h C12 urinary excretion was around 5% of the administered amount. The percentage of C12 oxidized was 81.7 +/- 9.5% (mean +/- S.D.) of administered amount as estimated from the area under the curve of measured (14)CO(2) expiration rate. C12 kinetics was described by assuming a single compartment. A saturable rate of C12 tissue uptake (model A) and a linear rate of tissue uptake (model B) were considered. The kinetics of CO(2) produced by C12 oxidation was described by a fast pathway acting in parallel to a slow pathway modeled by first order kinetics. Parameters of model B were estimated for each subject, whereas model A was identified by fitting the pooled data of all subjects. On the basis of estimates obtained from model B, an average calorie delivery of 500 kcal/day was predicted in the plateau phase for the infusion rate of our experiments. When estimated from model A, the maximal rate of tissue uptake was 0.38 +/- 0.08 mmol/min, with a maximal calorie delivery of 750 kcal/day. These results appear promising for C12 utilization in parenteral nutrition, because C12 elimination with urine is low, whereas tissue uptake and oxidation are rather efficient. PMID- 10688597 TI - Central administration of methamphetamine synergizes with metabolic inhibition to deplete striatal monoamines. AB - These studies examined, in vivo, the effect of local intrastriatal perfusion of methamphetamine (MA) on dopamine (DA) and glutamate release in relation to changes in striatal DA and serotonin (5-HT) content measured 1 week after treatment. Interactions between the inhibition of energy metabolism and the direct perfusion of MA on long-term decreases in DA and 5-HT content also were investigated. MA (100 microM), the succinate dehydrogenase inhibitor malonate, or the combination of MA and malonate was reverse-dialyzed into the striatum for 8 h. The continuous local perfusion of MA alone increased DA release by 30-fold, similar to that seen after systemic administration, but did not increase glutamate or body temperature, and did not deplete neurotransmitter content. Malonate perfusion increased both DA and glutamate overflow, and dose dependently decreased DA content. 5-HT content was not as affected by malonate perfusions (200 mM malonate depleted DA by 66% and 5-HT by 40%). When MA was coperfused with 200 mM malonate, DA content was reduced by 80% and to a greater extent compared with malonate alone. Coperfusion of MA and 200 mM malonate did not enhance 5-HT loss. Overall, the present findings provide evidence that energy metabolism plays an important role in MA toxicity and that striatal dopaminergic terminals are more vulnerable than 5-HT terminals to damage after metabolic stress. PMID- 10688598 TI - Structure-pharmacokinetics relationship of series of aminosteroidal neuromuscular blocking agents in the cat. AB - To obtain more insight in the relationship between physicochemical properties of neuromuscular blocking agents (NMBAs) and their pharmacokinetic characteristics, a series of 12 aminosteroidal NMBAs, supplemented with data on five related NMBAs from the literature, was investigated in anaesthetized cats. After i.v. bolus injection, plasma concentration decreased very rapidly, showing a biphasic pattern, with half-lives ranging from 0.4 to 1.4 min, and from 3 to 10 min, respectively. Clearance was in the range from 24 to 58 ml. min(-1). kg(-1). Compounds containing an acetyl-ester group at position 3 were partly metabolized to the 3-OH derivative. The urinary excretion of the parent drug and metabolites amounted to <10% for each of the compounds. The parent drugs were excreted in large amounts into bile, along with smaller amounts of 3-OH derivatives. The terminal half-life of the urinary and biliary excretion rate were markedly longer than the apparent terminal half-life in plasma, ranging from 11 to 40 min, and from 119 to 489 min in urine and bile, respectively. Lipophilicity of the NMBAs, expressed as the partition coefficient octanol/Krebs (log P), was found to be correlated positively with unbound plasma clearance and unbound initial plasma clearance, and negatively with plasma half-life, volume of distribution at steady state, and mean residence time. The increase of the unbound plasma clearance with increasing lipophilicity is counteracted by the concurrent increase in plasma protein binding. PMID- 10688599 TI - Inhibitors of phospholipase C prevent glutamate neurotoxicity in primary cultures of cerebellar neurons. AB - The role of phospholipase C in the molecular mechanism of glutamate neurotoxicity was assessed in primary cultures of cerebellar neurons. It is shown that 1-[6 [[(17b)-3-methoxyestra-1,3, 5(10)-trien-17-yl]amino] hexyl]-1H-pyrrole-2,5-dione (U-73122) and 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphorylcholine (Et-18 OCH(3)), two agents that inhibit phospholipase C, prevent glutamate and N-methyl D-aspartic acid (NMDA) neurotoxicity. It is shown that both compounds prevent glutamate neurotoxicity at concentrations lower than those required to inhibit carbachol-induced hydrolysis of inositol phospholipids. In contrast, it was a good correlation between the concentrations of U-73122 and Et-18-OCH(3) required to inhibit NMDA-induced hydrolysis of phospholipids and those required to prevent glutamate and NMDA neurotoxicity. NMDA-induced hydrolysis of phospholipids is inhibited by nitroarginine, an inhibitor of nitric-oxide synthase, and is mimicked by the nitric oxide-generating agent S-nitroso-N-acetylpenicillamine. The results reported indicate that glutamate neurotoxicity would be mediated by activation of NMDA receptors, leading to activation of nitric-oxide synthase and increased formation of nitric oxide, which results in increased activity of phospholipase C. Inhibition of phospholipase C by U-73122 or Et-18-OCH(3) prevents glutamate-induced neuronal death. PMID- 10688600 TI - M(2) and M(4) receptor knockout mice: muscarinic receptor function in cardiac and smooth muscle in vitro. AB - Peripheral muscarinic receptors play key roles in the control of heart rate and smooth muscle activity. In this study, bradycardic and smooth muscle contractile responses to the muscarinic agonist carbamylcholine were compared in isolated tissues from M(2) and M(4) muscarinic receptor knockout mice and their wild-type littermates. Carbamylcholine (1 x 10(-8)-3 x 10(-5) M) produced similar concentration-dependent bradycardia in spontaneously beating atria from M(4) receptor knockout and wild-type control mice. In contrast, carbamylcholine did not produce bradycardia in atria derived from M(2) receptor knockout mice, whereas such atria were responsive to adenosine-induced bradycardia. Carbamylcholine-induced contractile responses were similar in stomach fundus, urinary bladder, and tracheal preparations from M(4) receptor knockout mice and their wild-type littermates for each tissue (-logEC(50) values ranging from 6.20 +/- 0.10 to 6.76 +/- 0.08), suggesting that M(4) receptors do not participate in smooth muscle contraction in these tissues. In contrast, approximately 2-fold higher carbamylcholine concentration was required for contraction of stomach fundus, urinary bladder, and trachea from M(2) receptor knockout mice (-logEC(50) = 6.39 +/- 0.05, 6.07 +/- 0.06, and 6.27 +/- 0.12, respectively) than from wild type littermates (-logEC(50) = 6.68 +/- 0.07, 6.27 +/- 0.07, and 6.56 +/- 0.06, respectively). Furthermore, the affinity of the M(2) "selective" receptor antagonist AF-DX116 in inhibiting carbamylcholine-induced smooth muscle contraction was significantly reduced in M(2) receptor knockout mice compared with tissues from wild-type littermates. Collectively, these results provide direct and unambiguous evidence that M(2) receptors mediate muscarinic receptor induced bradycardia and play a role in smooth muscle contractility, whereas M(4) receptors are not involved in stomach fundus, urinary bladder, or tracheal contractility. PMID- 10688601 TI - Cloning and pharmacological characterization of the rat CB(2) cannabinoid receptor. AB - Many of the pharmacological effects of Delta(9)-tetrahydrocannabinol are mediated through CB(1) and CB(2) cannabinoid receptors. However, with the discovery of endogenous cannabinoids, some discrepancies have arisen. Furthermore, unlike the CB(1) receptor, the sequences of the mouse and human CB(2) receptor are divergent, raising the possibility of species specificity. The gene for the rat CB(2) receptor was cloned, expressed, and its properties compared with those of mouse and human CB(2) receptors. Sequence analysis of the coding region of the rat CB(2) genomic clone indicates 90% nucleic acid identity (93% amino acid identity) between rat and mouse and 81% nucleic acid identity (81% amino acid identity) between rat and human. The rat CB(2) receptor was stably expressed in human embryonic kidney-293 cells to examine its pharmacology. The rat CB(2) showed low affinity for anandamide, an endogenous ligand shown to act at the CB(1) receptor. In contrast, high-affinity binding for SR144528 (CB(2)-selective antagonist) as well as several cannabinoid receptor agonists was observed. Coupling to adenylate cyclase was observed. Aspects of the pharmacology of palmitoylethanolamide were also examined. It bound to CB(1) and CB(2) receptors with low affinity and stimulated GTPgammaS binding in the cerebellum and CB(2) expressing cell lines with low potency. The data in this study suggest that the discrepancies in affinities between rat and human may represent species differences. The rat CB(2) receptor genomic clone will be a useful tool for studying the function and regulation of CB(2) in rats. PMID- 10688602 TI - 6-NO(2)-norepinephrine increases norepinephrine release and inhibits norepinephrine uptake in rat spinal synaptosomes. AB - Nitric oxide has been shown to react under physiologic conditions with norepinephrine (NE) to produce 6-nitro-norepinephrine (6-NO(2)-NE), a compound that enhances NE release in the brain. Previous studies suggest that 6-NO(2)-NE is formed in the spinal cord and stimulates spinal NE release to produce analgesia. The purpose of the current studies was to examine the mechanisms by which 6-NO(2)-NE stimulates NE release in the spinal cord. Crude synaptosomes were prepared from spinal cords of male Sprague-Dawley rats and loaded with [(3)H]NE. Incubation of synaptosomes with 6-NO(2)-NE resulted in a release of NE, with a threshold of 1 microM 6-NO(2)-NE and a maximum effect of 30% fractional release. NE transporter inhibitors desipramine and nomifensine blocked NE release from 6-NO(2)-NE, and desipramine exhibited an IC(50) of 9.6 microM. NE release from 6-NO(2)-NE was dependent on external Na(+), but not Ca(2+) or the activity of guanylate cyclase. 6-NO(2)-NE also blocked uptake of [(3)H]NE into synaptosomes, with an IC(50) of 8.3 microM. These data are consistent with a direct action of 6-NO(2)-NE on noradrenergic terminals in the spinal cord to release NE. This action is independent of guanylate cyclase activation, and most likely shares a common mechanism with classic monoamine releasers such as amphetamine that cause direct release of NE from vesicles into the nerve terminal cytoplasm, leading to extracellular release by reverse transport. PMID- 10688603 TI - Modulation of cellular calcium by sigma-2 receptors: release from intracellular stores in human SK-N-SH neuroblastoma cells. AB - Human SK-N-SH neuroblastoma cells expressed sigma-1 and sigma-2 receptors with similar pharmacological profiles to those of rodent-derived tissues, although sigma-2 receptors exhibited some affinity differences that might suggest heterogeneity or species differences. Structurally diverse sigma ligands produced two types of increases in intracellular (cytosolic) Ca(2+) concentration ([Ca(2+)](i)) in these cells. CB-64D, CB-64L, JL-II-147, BD737, LR172, BD1008, haloperidol, reduced haloperidol, and ibogaine all produced an immediate, dose dependent, and transient rise in [Ca(2+)](i). Sigma-inactive compounds structurally similar to the most active sigma ligands and ligands for several neurotransmitter receptors produced little or no effect. The high activity of CB 64D and ibogaine (sigma-2-selective ligands) compared with the low activity of (+)-pentazocine and other (+)-benzomorphans (sigma-1-selective ligands), in addition to enantioselectivity for CB-64D over CB-64L, strongly indicated mediation by sigma-2 receptors. The effect of CB-64D and BD737 was blocked by the sigma antagonists BD1047 and BD1063, further confirming specificity as a receptor mediated event. The transient rise in [Ca(2+)](i) occurred in the absence of extracellular Ca(2+) and was completely eliminated by pretreatment of cells with thapsigargin. Thus, sigma-2 receptors stimulate a transient release of Ca(2+) from the endoplasmic reticulum. Prolonged exposure of cells to sigma-receptor ligands resulted in a latent and sustained rise in [Ca(2+)](i), with a pharmacological profile identical to that of the transient rise. This sustained rise in [Ca(2+)](i) was affected by neither the removal of extracellular Ca(2+) nor thapsigargin pretreatment, suggesting latent sigma-2 receptor-induced release from thapsigargin-insensitive intracellular Ca(2+) stores. Sigma-2 receptors may use Ca(2+) signals in producing cellular effects. PMID- 10688604 TI - Peroxynitrite, a two-edged sword in post-ischemic myocardial injury-dichotomy of action in crystalloid- versus blood-perfused hearts. AB - Peroxynitrite (ONOO(-)) is widely recognized as a mediator of NO. toxicity, but recent studies have indicated that this compound may also have physiologic activity and induces vascular relaxation as well as inhibition of platelet aggregation and neutrophil adhesion. The present experiment was designed to determine whether ONOO(-) may exert different effects on postischemic myocardial injury in a crystalloid perfusion environment versus a blood perfusion environment and, if it does, to clarify the mechanisms causing any differences. In Krebs-Henseleit buffer-perfused rabbit hearts, administration of ONOO(-) at the onset of reperfusion enhanced myocardial injury in a concentration-dependent fashion with a significant effective concentration of 30 microM. In contrast, in blood-perfused hearts, administration of ONOO(-) (1 to 30 microM) significantly attenuated postmyocardial injury as evidenced by improved cardiac function recovery, preserved endothelial function, decreased myocardial creatine kinase loss, and reduced necrotic size. The minimal and maximal protective concentrations were determined to be 1 and 3 microM, respectively. When a high concentration of ONOO(-) (i.e., 100 microM) was administered, a detrimental effect was observed. Administration of ONOO(-) decreased neutrophil accumulation in the ischemic-reperfused myocardial tissue in a concentration-dependent manner in blood-perfused hearts and inhibited neutrophil adhesion to cultured endothelial cells exposed to hypoxia/reoxygenation. Taken together, these results demonstrate that ONOO(-) may act as a "double-edged sword" in postischemic myocardial injury. This compound is directly toxic to the cardiac tissue at a relatively high concentration, but it can indirectly protect myocardial cells from neutrophil-induced injury at a much lower concentration. PMID- 10688605 TI - c-Myc antisense limits rat liver regeneration and indicates role for c-Myc in regulating cytochrome P-450 3A activity. AB - Expression of c-myc protein is associated with cell proliferation. The present study uses antisense oligomers to inhibit c-myc expression in the regenerating rat liver after 70% partial hepatectomy (PH). Antisense phosphorodiamidate morpholino oligomers (novel DNA analogs) were administered i.p. immediately after surgery to block expression of c-myc within the first 24 h after PH. A 20-mer PMO complimentary to the c-myc mRNA at the translation start site was an effective sequence (AVI-4126, 5'-ACGTTGAGGGGCATCGTCGC-3'). A single i.p. dose of 0.5 mg/kg AVI-4126 caused reduction of the regenerating liver c-myc protein in a sequence specific and dose-dependent manner. Inhibition of c-myc expression resulted in reduction of proliferating cell nuclear antigen and arrested cells in the G(0)/G(1) phase of the cell cycle. The ratio of G(2):G(0) cell populations in the regenerating liver 24 h after PH dropped from 29.1 in saline vehicle-treated rats to 18.0 in rats treated with 2.5 mg/kg AVI-4126. The expression of cell cycle checkpoint protein p53 was inhibited with increasing doses of AVI-4126, but expression of p21(waf-1) was unaffected. The activity of cytochrome P-450 3A2 (CYP3A2) was evaluated by immunoblot analysis and erythromycin N-demethylation. AVI-4126 did not alter CYP3A activity in nonhepatectamized animals but showed a dose-dependent decrease in PH rats. We conclude that AVI-4126, antisense oligomer to c-myc, can reduce cell proliferation in the regenerating rat liver. Furthermore, inhibition of c-myc may indirectly influence the expression of CYP3A. PMID- 10688606 TI - Sustained reduction in myocardial reperfusion injury with an adenosine receptor antagonist: possible role of the neutrophil chemoattractant response. AB - Recent studies have demonstrated that three membrane-permeant A(1) receptor antagonists reduced infarct size in a model of ischemia followed by brief reperfusion. However, it was not determined whether cardioprotection was mediated by nonspecific intracellular effects of these highly lipophilic drugs and whether the antagonists only delayed myocardial necrosis without affecting the ultimate infarct size. In the present study, closed-chest dogs were subjected to 90 min of left anterior descending coronary artery occlusion and 72 h of reperfusion and received either a nonmembrane-permeant adenosine receptor blocker that is devoid of direct intracellular effects and is 6-fold selective for the A(1) receptor [1, 3-dipropyl-8-p-sulfophenylxanthine (DPSPX); n = 11] or vehicle (n = 12). DPSPX was administered as three 200-mg boluses 60 min before and 30 and 120 min after reperfusion. The area of necrosis was determined histologically and expressed as a percentage of the area at risk. Baseline predictors of infarct size were similar in the two groups. The ratio of the area of necrosis to the area at risk was less in the DPSPX group (17.8 +/- 4.3% versus 35.0 +/- 1.9%; P =. 012), and DPSPX improved regional ventricular function. Under both basal and stimulated (formyl-Met-Leu-Phe) conditions, suspensions of human neutrophils generated extracellular adenosine levels (approximately 50 nM) sufficient to activate A(1) receptors. Moreover, both DPSPX and 1,3-dipropyl-8-cyclopentylxanthine, a selective A(1) receptor antagonist, significantly reduced the chemoattractant response of neutrophils to formyl-Met-Leu-Phe. We conclude that blockade of A(1) adenosine receptors attenuates myocardial ischemic/reperfusion injury, possibly in part by decreasing the chemoattractant response of neutrophils. PMID- 10688607 TI - A mibefradil metabolite is a potent intracellular blocker of L-type Ca(2+) currents in pancreatic beta-cells. AB - It has been shown that mibefradil (Ro 40-5967) exerts a selective inhibitory effect on T-type Ca(2+) currents, although at higher concentrations it can antagonize high voltage-activated Ca(2+) currents. The action of mibefradil on Ca(2+) channels is use- and steady-state-dependent and the binding site of mibefradil on L-type Ca(2+) channels is different from that of dihydropyridines. By using conventional whole-cell and perforated patch-clamp techniques, we showed that mibefradil has an inhibitory effect on both T- and L-type Ca(2+) currents in insulin-secreting cells. However, the effect on L-type Ca(2+) currents was time dependent and poorly reversible in perforated patch-clamp experiments. By using mass spectrometry, we demonstrated that mibefradil accumulates inside cells, and furthermore, a metabolite of mibefradil was detected. Intracellular application of this metabolite selectively blocked the L-type Ca(2+) current, whereas mibefradil exerted no effect. This study demonstrates that mibefradil permeates into cells and is hydrolyzed to a metabolite that blocks L-type Ca(2+) channels specifically by acting at the inner side of the channel. PMID- 10688608 TI - Radiotelemetric evaluation of hemodynamic effects of long-term ethanol in spontaneously hypertensive and Wistar-Kyoto rats. AB - This study determined the hemodynamic effects of chronic ethanol in telemetered freely moving age-matched spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. Changes in blood pressure (BP), heart rate (HR), and plasma norepinephrine (as index of sympathetic activity) were evaluated in pair-fed rats receiving liquid diet with or without ethanol (5%, w/v) for 12 weeks. The SHRs exhibited higher baseline BP and lower HR compared with WKY rats. When normalized for body weight, daily ethanol intake was higher in SHRs compared with WKY rats. However, blood ethanol concentration was similar except for a higher level in SHRs at weeks 7 through 9. Ethanol had no effect on BP in WKY rats but caused decreases in BP in SHRs that reached a maximum (approximately 30 mm Hg) at week 5 and remained thereafter. Ethanol also caused reductions in the BP variability and the circadian fluctuations in BP in SHRs but not in WKY rats. Plasma norepinephrine levels were elevated by ethanol in WKY rats, but not in SHRs. The HR was not affected by ethanol in SHRs and showed increases in WKY rats. These findings suggest that chronic ethanol feeding differentially affects BP in SHRs (hypotension) and WKY rats (no effect). The lack of a hypotensive response to ethanol in WKY rats may relate, at least partly, to the associated sympathoexcitation. The present study used the telemetry technique for BP measurement, which eliminates the confounding and stressful effects of other conventional techniques. PMID- 10688609 TI - D(2), but not D(1) dopamine receptor agonists potentiate cannabinoid-induced sedation in nonhuman primates. AB - In primates, CB(1) cannabinoid receptor agonists produce sedation and psychomotor slowing, in contrast to behavioral stimulation produced by high doses of dopamine receptor agonists. To investigate whether dopamine agonists attenuate the sedative effects of a cannabinoid agonist in monkeys, we compared the effects of D(1) or D(2) dopamine receptor agonists on spontaneous behavior in three to six cynomolgus monkeys (Macaca fasicularis) alone and after administration of a low dose of the CB(1) agonist levonantradol. Alone, the CB(1) cannabinoid receptor agonist levonantradol (0.01-0. 3 mg/kg) induced sedation, ptosis, and decreased locomotor and general activity. Alone, D(2)-type dopamine agonists quinelorane (0. 001-1.0 mg/kg; n = 4) or pergolide (0.01-1.0 mg/kg) or a D(1) dopamine agonist 6-chloro-7,8-dihydroxy-1-phenyl-2,3,4, 5-tetrahydro-3-allyl-[1H]-3 benzazepine (0.3-3.0 mg/kg) produced either no effect or promoted hyperactivity. Thirty minutes after administration of a threshold dose of levonantradol (0.03 mg/kg), D(2)-type agonists, but not the D(1) agonist, precipitated marked sedation, ptosis, and decreased general activity and locomotor activity. These data inducate the following: 1) D(2,) but not D(1) dopamine agonists, potentiate sedation in monkeys treated with a CB(1) cannabinoid agonist, at doses of agonists that alone do not produce sedation; 2) the threshold dose for cannabinoid-induced sedation is reduced by D(2) agonists, but not by a D(1) dopamine agonist, differentiating D(1) and D(2) dopamine receptor linkage to cannabinoid receptors; and 3) modulation of D(2) dopamine receptor activity by a nonsedating dose of a cannabinoid agonist has implications for the pathophysiology and treatment of dopamine-related neuropsychiatric disorders and drug addiction. Cannabinoid agonists and D(2) dopamine agonists should be combined with caution. PMID- 10688610 TI - Carrier-mediated uptake of the endogenous cannabinoid anandamide in RBL-2H3 cells. AB - Anandamide (N-arachidonylethanolamide) is an endogenous cannabinoid that mimics the pharmacologic effects of Delta(9)-tetrahydrocannabinol, the major bioactive substance in marijuana. Anandamide appears to be synthesized, released, and inactivated by mechanisms similar to those for other neurotransmitters. Of interest to the present studies are reports that anandamide undergoes carrier mediated uptake into neuronal or glial cells after release, followed by rapid intracellular degradation by the intracellular fatty acid amidohydrolase. In addition to effects in the brain, anandamide has multiple effects in the periphery, particularly on cells of the immune system that express both a peripheral cannabinoid receptor and amidohydrolase enzyme. We have performed a detailed characterization of anandamide uptake in the cognate mast cell line RBL 2H3 to test the hypothesis that the uptake system in peripheral cells is also carrier-mediated and functionally similar to that observed in the central nervous system. RBL-2H3 cells exhibited robust, saturable transport of [(3)H]anandamide that was both time- and temperature-sensitive. This transport activity was not dependent on extracellular ion gradients for uptake and was inhibited selectively by other fatty acid-derived molecules, anandamide congeners, and the psychoactive cannabinoids such as Delta(9)-tetrahydrocannabinol. We conclude that anandamide transport in the RBL-2H3 cells is carrier-mediated, and uptake in peripheral cells is functionally and pharmacologically identical with that observed in neurons and astrocytes. PMID- 10688611 TI - Metabolite anion carriers mediate the uptake of the anionic drug fluorescein in renal cortical mitochondria. AB - The fluorescent organic anion fluorescein (FL) accumulates in proximal tubular cells of the kidney during renal secretion. In freshly isolated and permeabilized proximal tubular cells, the uptake was reduced but still sensitive to probenecid, suggesting a concentrative mechanism that is associated with intracellular compartments. Previous studies have shown that one of these compartments may be mitochondrial. In this study, we further investigated the transport characteristics of FL in isolated rat kidney cortex mitochondria. Mitochondrial uptake of 100 microM FL was rapid, with an initial rate of 60 pmol/mg protein.min, and reached equilibrium after 5 min. To characterize the transport system(s) involved, FL uptake was studied in the absence and presence of substrates or inhibitors specific for the various mitochondrial anion carriers. Phenylsuccinate (10 mM), an inhibitor of the alpha-ketoglutarate carrier, reduced uptake significantly with a maximum inhibition of 33% and an inhibitory constant (-log IC(50)) of 4.0 +/- 0.4 (P <.05). The apparent K(m) for the phenylsuccinate corrected FL uptake was 1.3 +/- 0.3 mM with a V(max) of 260 +/- 26 pmol/mg protein.15 s. Substrates for the tricarboxylate and glutamate-aspartate carriers significantly reduced the uptake of 100 microM FL with -log IC(50) values of 4.6 +/- 0.4 (citrate), 5.5 +/- 0.3 (glutamate), and 4.1 +/- 0.4 (aspartate). Substrates for the monocarboxylate and dicarboxylate carriers were without effect. The anionic drugs, valproate, indomethacin, and salicylate, significantly reduced FL uptake, whereas cephaloglycin and cephaloridine had no effect. Finally, a combination of phenylsuccinate, glutamate, and citrate reduced the uptake by 66%, indicating that at least three metabolite carriers contribute concomitantly to intramitochondrial FL transport. PMID- 10688612 TI - Effects of intracavernous administration of selective antagonists of alpha(1) adrenoceptor subtypes on erection in anesthetized rats and dogs. AB - The proerectile properties of three novel alpha(1)-adrenoceptor (alpha(1)-ADR) antagonists with different profiles of selectivity for the alpha(1)-ADR subtypes have been evaluated in anesthetized rats and dogs on intracavernous (IC) injection, in comparison with prazosin and phentolamine. In rats, the tested compounds decreased blood pressure (BP) and increased IC pressure (ICP), as well as the ratio ICP/BP. Rec 15/2841 (alpha(1a)- plus alpha(1L)-ADR-selective antagonist) and Rec 15/2615 (alpha(1b)-ADR selective) were the most potent compounds. The ICP/BP ratios calculated after injection of Rec 15/3039 (alpha(1d) ADR selective) were not markedly different from those observed after vehicle injection. Prazosin and phentolamine proved poorly active, their main effect being hypotension. Approximate ED(25) values (dose of compound in micrograms inducing 25% increase of ICP/BP ratio) were Rec 15/2615 (22 microgram/kg)>= Rec 15/2841 (29 microgram/kg) > prazosin (136 microgram/kg) > phentolamine (1298 microgram/kg) > Rec 15/3039 (9600 microgram/kg). Submaximal stimulation of the cavernous nerve elicited an ICP rise whose amplitude was not altered by Rec compounds. In contrast, prazosin and phentolamine decreased this ICP rise. All compounds but 15/3039 induced significant increase of the ICP/BP ratio in dogs. Rec 15/2615 proved to be the most interesting compound, inducing significant increases of ICP/BP at doses practically devoid of effects on BP. The rank order of potency in dog in increasing the ICP/BP ratio was similar to that observed in rats. Only at the highest doses tested, all compounds, except Rec 15/3039, decreased the ICP rise elicited by submaximal stimulation of the cavernous nerve. Our data demonstrate that the alpha(1b)- and alpha(1L)-ADR subtypes are functionally relevant for the erectile function in these models, and that alpha(1b)- and/or alpha(1L)-ADR subtypes selective antagonists could represent a real advantage in erectile dysfunction therapy. PMID- 10688613 TI - Possible mechanism of hepatocyte injury induced by diphenylamine and its structurally related nonsteroidal anti-inflammatory drugs. AB - Diphenylamine is a common structure of nonsteroidal anti-inflammatory drugs (NSAIDs) to uncouple mitochondrial oxidative phosphorylation and to cause a decrease in hepatocellular ATP content and hepatocyte injury. The mechanism for acute cell injury induced by diphenylamine and its structurally related NSAIDs was investigated with rat liver mitochondria and freshly isolated hepatocytes, focusing on the relation to the uncoupling of oxidative phosphorylation. Incubation of mitochondria with diphenylamine as well as mefenamic acid and diclofenac caused pseudoenergetic mitochondrial swelling, indicating that these compounds induce mitochondrial membrane permeability transition. Diphenylamine also caused changes in safranine-binding spectra to mitochondria that was energized by succinate oxidation. This spectral shift indicates the loss of mitochondrial membrane potentials, which is known as one of the characteristics for uncouplers of oxidative phosphorylation, and also was caused by mefenamic acid and diclofenac. Incubation of hepatocytes with mefenamic acid, diclofenac, and diphenylamine diminished cellular ATP content, followed by leakage of lactose dehydrogenase from hepatocytes. Fructose, a low K(m) substrate for glycolysis, partially protected against the ATP depletion and hepatocyte injury induced by these compounds. Further addition of oligomycin, which blocks ATPase, pronounced the protection against cell injury. These results suggested that decreases in cellular ATP content, mainly caused by uncoupling of mitochondrial oxidative phosphorylation, were responsible for acute hepatocyte injury induced by diphenylamine and structurally related NSAIDs. PMID- 10688614 TI - Interleukin-13 modulates collagen homeostasis in human skin and keloid fibroblasts. AB - Interleukin (IL)-13 has been implicated in the pathogenesis of various diseases characterized by fibrosis. We describe the effects of IL-13 on collagen homeostasis from normal (NF) and keloid (KF) fibroblasts and compare these effects with those of IL-4 and transforming growth factor (TGF)-beta(1). Total collagen generation was up-regulated in NF after 48 h of stimulation by IL-13; in KF, IL-13 stimulated a more rapid collagen response. The kinetics and magnitude of collagen generation induced by IL-13 were equivalent to those induced by similar concentrations of IL-4 and TGF-beta(1). Collagen type I production paralleled total collagen generation from both NF and KF; however, IL-4-induced collagen type I and total collagen production from KF was more transient than that induced by either IL-13 or TGF-beta(1). Procollagen 1alpha1 gene expression was induced in KF by stimulation with IL-13 for 24 h. Moreover, IL-13 was unique among these three cytokines in its ability to induce gene expression for procollagen 3alpha1. Finally, IL-13 inhibited IL-1beta-induced matrix metalloproteinase (MMP)-1 and MMP-3 production and enhanced tissue inhibitor of metalloproteinase (TIMP)-1 generation from NF; although similar effects were observed with IL-4, TGF-beta(1) transiently enhanced MMP-1 and MMP-3 generation without effecting TIMP-1. In KF, IL-13 and IL-4 inhibited MMP-3, whereas TGF beta(1) enhanced MMP-3; TIMP-1 was unaffected by any of the three cytokines. These data demonstrate both the profibrotic effects of IL-13 on collagen homeostasis and the potential differential regulation of collagen homeostasis in fibroblast subtypes by IL-13. PMID- 10688615 TI - Ca(2+) mobilization evoked by chloroform in Madin-Darby canine kidney cells. AB - The effect of chloroform on Ca(2+) mobilization in Madin-Darby canine kidney cells was examined by using Fura-2 as a Ca(2+) probe. Chloroform (24-248 mM) concentration dependently increased intracellular Ca(2+) concentration ([Ca(2+)](i)). Ca(2+) removal inhibited the Ca(2+) signals evoked by 93 to 248 mM chloroform by reducing both the initial rise and the sustained phase. In Ca(2+) free medium, pretreatment with 93 mM chloroform abolished the Ca(2+) release induced by 1 microM thapsigargin, an endoplasmic reticulum Ca(2+) pump inhibitor, and partially reduced the Ca(2+) release induced by 2 microM carbonylcyanide m chlorophenylhydrazone, a mitochondrial uncoupler. Pretreatment with carbonylcyanide m-chlorophenylhydrazone and thapsigargin to deplete the Ca(2+) stores in mitochondria and the endoplasmic reticulum, respectively, only partially inhibited chloroform-induced Ca(2+) release. This suggests that chloroform released Ca(2+) from multiple internal pools. The addition of 3 mM Ca(2+) increased [Ca(2+)](i) after pretreatment with 93 mM chloroform in Ca(2+) free medium. La(3+) (1 mM) partially inhibited the [Ca(2+)](i) increase induced by 93 mM chloroform. Chloroform (93 mM)-induced Ca(2+) release was not altered when the formation of inositol-1,4,5-trisphosphate was abolished by U73122 (2 microM), a phospholipase C inhibitor, but was inhibited by 90% by inhibition of phospholipase A(2) with 40 microM aristolochic acid. Collectively, we found that 93 mM chloroform increased [Ca(2+)](i) in Madin-Darby canine kidney cells by releasing Ca(2+) from multiple stores in a manner independent of the formation of inositol-1,4,5-trisphosphate, followed by Ca(2+) entry from external medium. Other solvents, such as ethanol, methanol, and DMSO, did not affect the resting [Ca(2+)](i) at a concentration of 248 mM. PMID- 10688616 TI - Probable involvement of the 5-hydroxytryptamine(4) receptor in methotrexate induced delayed emesis in dogs. AB - Delayed emesis in cancer patients undergoing chemotherapy remains a significant problem. The pathogenesis of delayed emesis is still obscure. It was recently demonstrated that methotrexate (MTX), an anticancer drug, evoked delayed emesis in dogs in a manner similar to its actions in humans. We evaluated the antiemetic activity of FK1052, a potent antagonist for both the 5-hydroxytryptamine (HT)(3) and 5-HT(4) receptors, on delayed emesis induced by MTX in beagle dogs. Animal behavior was recorded for 3 days using a video camera. Delayed emesis lasting up to 72 h was observed in dogs treated with MTX (2.5 mg/kg i.v.), but acute emesis did not occur. The following antiemetics, at the dose that prevents cisplatin induced acute emesis in dogs, were administered i.v. as multiple injections every 12 h during days 2 to 3. FK1052 (1 and 3.2 mg/kg) significantly reduced the emetic episodes caused by MTX, whereas ondansetron (1 mg/kg), a selective 5-HT(3) receptor antagonist, was not effective. The emetic episodes induced by MTX were also inhibited by another 5-HT(3/4) receptor antagonist, tropisetron (1 mg/kg). CP-122,721 (0. 1 mg/kg), a potent selective tachykinin NK(1) receptor antagonist, significantly reduced the emetic responses to MTX. Copper sulfate-induced emesis in dogs was also prevented by FK1052, tropisetron, and CP-122,721 but not by ondansetron. FK1052, tropisetron, and ondansetron had negligible affinity for the NK(1) receptor at 1 microM. These results suggest that the 5-HT(4) receptor may be in part involved in the production of delayed emesis induced by MTX in dogs and that FK1052 may be a useful drug against both acute and delayed emesis induced by cancer chemotherapy. PMID- 10688617 TI - Induction of cytochrome P-450 1A2 by oxidized tryptophan in Hepa lclc7 cells. AB - Recent studies from this laboratory have demonstrated that L-tryptophan, after oxidation either by UV-irradiation or ozone, induces aryl hydrocarbon receptor (AhR) activation and binding of the liganded AhR complex to its specific DNA recognition site, thereby initiating transcription of the cytochrome P-450 1a1 (Cyp1a1) gene with concomitant increase of CYP1A1 protein and 7-ethoxyresorufin O deethylase activity in wild-type mouse hepatoma cells, Hepa lclc7 (Hepa-1), in culture. Temporary inhibition of protein synthesis by cycloheximide resulted in superinduction of oxidized tryptophan-inducible CYP1A1 mRNA, protein, and 7 ethoxyresorufin O-deethylase activity in Hepa-1 cells. In the present communication, the results obtained by immunoblot analyses with monoclonal CYP1A1/1A2 antibody (NIH 1-7-1) demonstrate that both UV- or ozone-oxidized tryptophan also induce CYP1A2 protein in Hepa-1 cells. CYP1A2 mRNA, detected by reverse transcription-polymerase chain reaction, was markedly induced in the UV- or ozone-oxidized tryptophan-treated cells. Temporary inhibition of protein synthesis by cycloheximide further induced oxidized tryptophan-inducible CYP1A2 mRNA as well as the protein in Hepa-1 cells. This is the first report demonstrating the induction of CYP1A2 mRNA and protein in Hepa-1 cells. PMID- 10688618 TI - Block of human heart hH1 sodium channels by amitriptyline. AB - Amitriptyline is a tricyclic antidepressant used to treat major depression and various neuropathic pain syndromes. This drug also causes cardiac toxicity in patients with overdose. We characterized the tonic and use-dependent amitriptyline block of human cardiac (hH1) Na(+) channels expressed in human embryonic kidney cells under voltage-clamp conditions. Our results show that, near the therapeutic plasma concentration of 1 microM, amitriptyline is an effective use-dependent blocker of hH1 Na(+) channels during repetitive pulses (approximately 55% block at 5 Hz). The tonic block for resting and for inactivated hH1 channels by amitriptyline (0.1-100 microM) yielded IC(50) values (50% inhibitory concentration) of 24.8 +/- 2.0 (n = 9) and 0.58 +/- 0.03 microM (n = 7), respectively. Substitution of phenylalanine with lysine at the hH1-F1760 position, a putative binding site for local anesthetics, eliminates the use dependent block by amitriptyline at 1 microM. The time constants of recovery from the inactivated-state amitriptyline block in hH1 wild-type and hH1-F1760K mutant channels are 8.0 +/- 0. 5 (n = 6) and 0.45 +/- 0.07 s (n = 6), respectively. A substitution at either hH1-F1760K or hH1-Y1767K significantly increases the IC(50) values for resting and inactivated states of amitriptyline, but the increase is much more pronounced with the hH1-F1760K mutation. Because these two residues were proposed to form a part of the local anesthetic binding site, we conclude that amitriptyline and local anesthetics interact with a common binding site. Furthermore, at therapeutic concentrations, the ability of amitriptyline to act as a potent use-dependent blocker of Na(+) channels may, in part, explain its analgesic actions. PMID- 10688619 TI - Effects of age on in vitro midazolam biotransformation in male CD-1 mouse liver microsomes. AB - To study age-related changes in drug metabolism, we examined the in vitro biotransformation of midazolam (MDZ), a human cytochrome P-450 (CYP) 3A substrate, using liver microsomes from three age groups of male CD-1 mice ranging from 6 weeks to 2 years old. MDZ was metabolized to two major products, alpha-OH- and 4-OH-MDZ, which were quantified by HPLC. For both metabolites, V(max) values were reduced in old livers (P <.05), while K(m) values did not change with age. The net intrinsic clearance (the sum of V(max)/K(m) for both pathways) also was reduced in the old animals (P <.05). The capacity of ketoconazole, a CYP3A inhibitor in humans, to inhibit the biotransformation of MDZ and of alprazolam, another human CYP3A substrate, did not differ significantly with age. At 100 microM alprazolam, 0.5 microM ketoconazole inhibited metabolite formation by >80%. At 30 microM MDZ, 2.5 microM ketoconazole impaired 4-OH-MDZ formation by 88%, whereas it reduced alpha-OH-MDZ formation by only 46%. Immunoinhibition studies with polyclonal anti-rat CYP3A1/2 and CYP2C11 antibodies confirmed that 4 OH-MDZ formation was largely CYP3A-dependent, while alpha-OH-MDZ formation was mediated by CYP3A and -2C isoforms. Western blot analysis revealed decreased microsomal content of CYP3A in old livers. Net intrinsic clearance of MDZ was correlated with total CYP3A content (P <.001). These results demonstrate a reduction in MDZ biotransformation in old male mice, which may be attributable, in part, to decreased CYP3A content in old livers. Changes in expression and activity of CYP2C isoforms also may contribute to age-related changes in MDZ biotransformation, but this requires more investigation. PMID- 10688620 TI - Polarized distribution of interleukin-1 receptors and their role in regulation of serotonin transporter in placenta. AB - We investigated the expression of interleukin-1 (IL-1) receptors and their involvement in the regulation of the serotonin transporter gene expression in human placenta. IL-1beta is an activator of the serotonin transporter gene expression in JAR human placental choriocarcinoma cells as demonstrated by an increase in the steady-state levels of the transporter mRNA and in serotonin transport activity. This activation is blocked by IL-1 receptor antagonist. Genistein also blocks the effect of IL-1beta, indicating involvement of tyrosine phosphorylation in the process. Treatment of JAR cells with IL-1beta activates mitogen-activated protein kinases and nuclear factor-kappaB. The nuclear factor kappaB that is responsive to IL-1beta in these cells is the p65 homodimer. Northern blot analysis and reverse transcription-polymerase chain reaction revealed that JAR cells and human placenta express type I and type II IL-1 receptors. The binding sites for (125)I-IL-1beta are localized predominantly in the maternal-facing brush border membrane of the syncytiotrophoblast. These results show that IL-1 in the maternal circulation is likely to play a critical role in the regulation of the serotonin transporter gene expression in the placenta. PMID- 10688621 TI - Disposition of methamphetamine and its metabolite amphetamine in brain and other tissues in rats after intravenous administration. AB - These studies characterized the concentration-time profile of (+)-methamphetamine [(+)-METH] and its metabolite (+)-amphetamine [(+)-AMP] in the brain and five other tissues after (+)-METH administration. Male Sprague-Dawley rats received a pharmacologically active (+)-METH i.v. bolus dose (1.0 mg/kg) or a nonpharmacologically active s.c. infusion (20 h at 1.2 mg/kg/day). Tissues (n = 3 per time point) were collected for more than four elimination half-lives in the i.v. group, or at a single steady-state time point (20 h) in the s.c. group. Based on data from the area under the concentration-time curves after i.v. dosing, the rank order of (+)-METH tissue accumulation was kidney > spleen > brain > liver > heart > serum with terminal elimination half-life values ranging from 53 to 66 min. (+)-METH concentrations were highest at the first measured time point (2 min) in all tissues except the spleen, which peaked at 10 min. The brain-to-serum concentration ratio rose from 7:1 at 2 min to a peak of 13:1 at 20 min before equilibrating to a constant value of 8:1 at 2 h. Following s.c. (+) METH dosing, the (+)-METH brain-to-serum concentration ratio was the same as the equilibrated ratio following i.v. dosing. (+)-AMP concentrations peaked at 20 min in all tissues before decaying with terminal elimination half-life values ranging from 68 to 75 min. Analysis of the area under the concentration-time curve molar amounts of (+)-AMP and (+)-METH showed that (+)-AMP accounted for approximately one-third of the drug tissue exposure over time. Thus, these data indicate the importance of both (+)-METH and (+)-AMP in pharmacological effects following i.v. (+)-METH administration. PMID- 10688622 TI - Targeting rat anti-mouse transferrin receptor monoclonal antibodies through blood brain barrier in mouse. AB - Drug targeting through the brain capillary endothelium, which forms the blood brain barrier (BBB) in vivo, may be achieved with peptidomimetic monoclonal antibodies that target peptide transcytosis systems on the BBB in vivo. Murine monoclonal antibodies to the rat transferrin receptor, such as the OX26 monoclonal antibody, are targeted through the BBB on the transferrin receptor in the rat. However, the present studies show the OX26 monoclonal antibody is not an effective brain delivery vector in mice. The emergence of transgenic mouse models creates a need for brain drug-targeting vectors for this species. Two rat monoclonal antibodies, 8D3 and RI7-217, to the mouse transferrin receptor were evaluated in the present studies. Both the RI7-217 and the 8D3 antibody had comparable permeability-surface area products at the mouse BBB in vivo. However, owing to a higher plasma area under the concentration curve, the mouse brain uptake of the 8D3 antibody was higher, 3.1 +/- 0.4% of injected dose [(ID)/g] compared with the brain uptake of the RI7 antibody, 1.6 +/- 0.2% ID/g, at 60 min after i.v. injection. Conversely, the mouse brain uptake of the OX26 antibody, which does not recognize the mouse transferrin receptor, was negligible, 0.06 +/- 0.01% ID/g. The RI7-127 antibody was more selective for brain because this antibody was not measureably taken up by liver. The capillary depletion technique demonstrated transcytosis of the RI7-217 antibody through the mouse BBB in vivo. The brain uptake of the 8D3 antibody was saturable, consistent with a receptor mediated transport process. In conclusion, these studies indicate rat monoclonal antibodies to the mouse transferrin receptor may be used for brain drug-targeting studies in mice such as transgenic mouse models. PMID- 10688623 TI - Reward and somatic changes during precipitated nicotine withdrawal in rats: centrally and peripherally mediated effects. AB - The negative affective aspects of nicotine withdrawal have been hypothesized to contribute to tobacco dependence. In the present studies in rats, brain stimulation reward thresholds, conditioned place aversions, and somatic signs of withdrawal were used to investigate the role of central and peripheral nicotinic acetylcholine and opioid receptors in nicotine withdrawal. Rats prepared with s.c. osmotic mini-pumps delivering 9.0 mg/kg/day nicotine hydrogen tartrate or saline were administered various doses of the nicotinic antagonists mecamylamine (s.c.), chlorisondamine (s. c. or i.c.v.), dihydro-beta-erythroidine (s.c.), or the opiate antagonist naloxone (s.c.). Nicotine-treated rats receiving mecamylamine or i.c.v. chlorisondamine exhibited elevated thresholds and more somatic signs than saline-treated rats. Nicotine-treated rats receiving s.c. chlorisondamine, at doses that do not readily cross the blood-brain barrier, exhibited more somatic signs than saline-treated rats with no threshold elevations. Naloxone administration produced threshold elevations and somatic signs only at high doses that induced similar magnitude effects in both nicotine- and saline-treated subjects. Mecamylamine or dihydro-beta-erythroidine administration induced conditioned place aversions in nicotine-treated rats but required higher doses than those needed to precipitate threshold elevations. In contrast, naloxone administration induced conditioned place aversions at lower doses than those required to precipitate threshold elevations and somatic signs. These data provide evidence for a dissociation between centrally mediated elevations in reward thresholds and somatic signs that are both centrally and peripherally mediated. Furthermore, threshold elevations and somatic signs of withdrawal appear to be mediated by cholinergic neurotransmission, whereas conditioned place aversions appear to be primarily mediated by the opioid system. PMID- 10688624 TI - The role of phosphodiesterase in mediating the effect of protein kinase C on cyclic AMP accumulation upon kappa-opioid receptor stimulation in the rat heart. AB - This study determined whether phosphodiesterase (PDE) was activated by protein kinase C (PKC) upon kappa-receptor stimulation, and if so, to identify the isozyme. We first studied the effects of trans-(+/-)-3,4-dichloro-N-methyl-N-(2 [1-pyrrolidinyl] cyclohexyl) benzeneacetamide methanesulphonate (U50,488H), a selective kappa-opioid receptor (OR) agonist, and phorbol-12-myristate-13-acetate (PMA), a PKC activator, on cAMP accumulation and PDE activity in rat ventricular myocytes when PKC and PDE were inhibited by respective inhibitors. Like PMA, U50,488H decreased the forskolin-stimulated cAMP accumulation and dose dependently stimulated the PDE activity, which were antagonized by 10(-6) M chelerythrine and bisindolylmaleimide I, selective PKC antagonists. In addition, 3-isobutyl-1-methylxanthine, a PDE inhibitor, dose-dependently attenuated the inhibition on forskolin-stimulated cAMP accumulation and abolished the stimulation on PDE activity by U50,488H and PMA. The observations suggest that PKC may enhance cAMP degradation through activating PDE upon kappa-OR stimulation. To identify the isozyme(s) mediating the effect of PKC upon kappa-OR stimulation, selective inhibitors were used. We found that 10(-5) M Ro-20-1724, a selective cAMP-specific PDE (PDE-IV) inhibitor, abolished the inhibitory effects of U50,488H and PMA, whereas 8-methoxymethyl-3-isobutyl-1-methylxanthine, erythro 9-(2-hydroxy-3-nonyl) adenine, cilostamide, and zaprinast, selective inhibitors of Ca(2+)/calmodulin-dependent PDE (PDE-I), cGMP-stimulated PDE (PDE-II), cGMP inhibited PDE (PDE-III), and cGMP-specific PDE (PDE-V), respectively, had no effect. Moreover, rolipram, another selective PDE-IV inhibitor, also dose dependently attenuated the inhibition on forskolin-stimulated cAMP accumulation and stimulation on PDE activity by U50,488H and PMA. In conclusion, this study has provided evidence for the first time that PKC and PDE-IV mediate the action of kappa-OR. PMID- 10688625 TI - Accelerated blood clearance and altered biodistribution of repeated injections of sterically stabilized liposomes. AB - Sterically stabilized liposomes are considered promising carriers of therapeutic agents because they can facilitate controlled release of the drugs, thereby reducing drug-related toxicity and/or targeted delivery of drugs. Herein, we studied the pharmacokinetics and biodistribution of repeated injections of radiolabeled polyethyleneglycol (PEG) liposomes. Weekly injections of (99m)Tc-PEG liposomes dramatically influenced the circulatory half-life in rats. Biodistribution 4 h after the second dose showed a significantly reduced blood content (from 52.6 +/- 3.7 to 0.6 +/- 0.1% injected dose (ID), P <.01) accompanied by a highly increased uptake in the liver (from 8.1 +/- 0.8 to 46.2 +/- 9.8%ID, P <.01) and in the spleen (from 2.2 +/- 0.2 to 5.3 +/- 0.7%ID, P <.01). At subsequent injections the effect was less pronounced: after the fourth dose, the pharmacokinetics of the radiolabel had almost returned to normal. The same phenomenon was observed in a rhesus monkey, but not in mice. The enhanced blood clearance of the PEG liposomes also was observed in rats after transfusion of serum from rats that had received PEG liposomes 1 week earlier, indicating that the enhanced blood clearance was caused by a soluble serum factor. This serum factor was a heat-labile molecule that coeluted on a size exclusion column with a 150-kDa protein. In summary, i.v. administration of sterically stabilized PEG liposomes significantly altered the pharmacokinetic behavior of subsequently injected PEG liposomes in a time- and frequency-dependent manner. The observed phenomenon may have important implications for the repeated administration of sterically stabilized liposomes for targeted drug delivery. PMID- 10688626 TI - Suppression of a high-affinity transport system for manganese in cadmium resistant metallothionein-null cells. AB - Cadmium is a hazardous heavy metal existing ubiquitously in the environment, but the mechanism of cadmium transport into mammalian cells has been poorly understood. Recently, we have established a cadmium-resistant cell line (Cd-rB5) from immortalized metallothionein-null mouse cells, and found that Cd-rB5 cells exhibited a marked decrease in cadmium uptake. To investigate the mechanism of altered uptake of cadmium in Cd-rB5 cells, incorporation of various metals was determined simultaneously using a multitracer technique. Cd-rB5 cells exhibited a marked decrease in manganese incorporation as well as that of cadmium. However, the reduced uptake of manganese was observed only at low concentrations, suggesting that a high-affinity component of the Mn(2+) transport system was suppressed in Cd-rB5 cells. Competition experiments and kinetic analyses revealed that low concentrations of Cd(2+) and Mn(2+) share the same high-affinity pathway for their entry into cells. The mutual competition of Cd(2+) and Mn(2+) uptake was also observed in HeLa, PC12, and Caco-2 cells. The highest uptake of Cd(2+) and Mn(2+) by parental cells occurred at neutral pH, suggesting that this pathway is different from a divalent metal transporter 1 that can transport various divalent metals including Cd(2+) and Mn(2+) under acidic conditions. These results suggest that a high-affinity Mn(2+) transport system is used for mammalian cellular cadmium uptake, and that the suppression of this pathway caused a marked decrease in cadmium accumulation in cadmium-resistant metallothionein-null cells. PMID- 10688627 TI - Benazeprilat disposition and effect in dogs revisited with a pharmacokinetic/pharmacodynamic modeling approach. AB - The pharmacokinetic disposition of benazeprilat, an angiotensin-converting enzyme (ACE) inhibitor (ACEI), was assessed with a nonlinear binding model in dogs. A single oral benazepril dose, a single i.v. benazeprilat dose, or a daily oral dose of benazepril for 14 consecutive days was administered. The activity of benazeprilat was assessed by measuring plasma ACE inhibition with an ex vivo assay. Benazeprilat data were fitted to equations corresponding to a monocompartmental model with a volume equal to the extracellular space ( approximately 0.2 l/kg) in which a fraction of benazeprilat was nonlinearily bound to ACE with both a saturable tissue and nontissue binding. The half-life of benazeprilat elimination determined from this physiologically based model was 39 +/- 6 min. The estimated maximal binding capacity of benazeprilat to ACE was approximately 23.5 nmol/kg, 90% of which was tissular. The estimated equilibrium constant of dissociation (K(d)) of benazeprilat to ACE was 2.7 to 4.5 nM. IC(50) values were one order of magnitude lower than K(d) values (i.e., approximately 0.27 nM). The nonlinear disposition of benazeprilat raised several issues and it was concluded that the benazeprilat concentration profile was only relevant to definition of an optimal dosage regimen if the appropriate kinetic model was used to interpret the plasma data. PMID- 10688628 TI - New insights on effect of kidney insufficiency on disposition of angiotensin converting enzyme inhibitors: case of enalapril and benazepril in dogs. AB - The influence of a renal injury on the disposition of benazeprilat, the active moiety of benazepril, and of enalaprilat, the active moiety of enalapril, two angiotensin-converting enzyme (ACE) inhibitors (ACEI), having different routes of elimination in dog was investigated during a mild renal insufficiency obtained by a nephrectomy-electrocoagulation method reducing glomerular filtration rate by approximately 50%. Plasma concentrations of the active moieties were analyzed with a physiologically based model taking into account the binding to ACE (high affinity, low capacity). An influence of renal insufficiency on enalapril disposition was shown with an increase in its plasma concentration, which was correlated to the reduction of the glomerular filtration rate. No such effect was evidenced for benazepril. With the physiologically based model analysis, it was shown that renal impairment led to an increase of the apparent benazeprilat clearance (260%), whereas that of enalaprilat was reduced to 40 to 55%. Renal insufficiency had no significant effect either on the apparent volume of distribution of each drug or on the binding parameters [i.e., maximal binding capacity (B(max)) and affinity (K(d))]. Enalaprilat and benazeprilat inhibitory action on ACE also was evaluated ex vivo. Similar patterns of inhibition were observed for both drugs. Renal injury had no significant influence on the overall effect of benazeprilat, whereas the inhibition effect of enalaprilat was significantly increased. It was concluded that renal insufficiency may have effects on the ACEI disposition but that the measurable active moiety plasma concentration is not the most appropriate endpoint to describe and interpret the consequence of a renal injury on ACEI. PMID- 10688629 TI - Inhibition by intracellular Mg(2+) of recombinant N-methyl-D-aspartate receptors expressed in Chinese hamster ovary cells. AB - Intracellular Mg(2+) (Mg(i)(2+)) inhibits the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors in cultured cortical neurons. To examine the effects of Mg(i)(2+) on recombinant NMDA receptors composed of subunit combinations found in cortical neurons, we expressed heteromeric receptors composed of NR1/NR2A and of NR1/NR2B subunits in Chinese hamster ovary (CHO) cells. We recorded whole-cell currents from the recombinant receptors in the absence and presence of Mg(i)(2+). The voltage dependence of control (0 Mg(i)(2+)) NMDA-activated currents obtained from CHO cells transfected with NR1/NR2A and with NR1/NR2B receptors showed outward rectification, a property that has been observed previously in native cortical NMDA receptors. The magnitude and voltage dependence of inhibition by Mg(i)(2+) of NMDA-activated currents were similar in CHO cells transfected with NR1/NR2A receptors, CHO cells transfected with NR1/NR2B receptors, and in cultured neurons expressing native NMDA receptors. These observations suggest that Mg(i)(2+) has uniform effects on the native NMDA receptors expressed in cortical neurons. Furthermore, inhibition by Mg(i)(2+) must not depend on intracellular factors or post-translational receptor modifications that are specific to neurons. Finally, the results indicate that the previously observed differences between whole-cell and outside-out patch measurements of Mg(i)(2+) inhibition could not result from poor control of voltage or Mg(i)(2+) concentration in the dendrites of neurons. The most likely alternative explanation is that patch excision causes an alteration in NMDA receptors that results in more effective inhibition by Mg(i)(2+). PMID- 10688630 TI - Regulation of extracellular concentrations of 5-hydroxytryptamine (5-HT) in mouse striatum by 5-HT(1A) and 5-HT(1B) receptors. AB - The ability of selective serotonin (5-HT) receptor agonists to reduce the extracellular concentration of 5-HT was examined in the striatum of awake, unrestrained mice by in vivo microdialysis. Systemic administration of either 8 OH-PIPAT (R-(+)-trans-8-hydroxy-2-[N-n-propyl-N-(3'-iodo-2'-propenyl)] aminotetralin), a novel 5-HT(1A) receptor agonist, or CP 94,253, a selective 5 HT(1B) receptor agonist, resulted in significant dose-related reductions of striatal 5-HT. The effect of 8-OH-PIPAT (1.0 mg/kg) was blocked by pretreatment with WAY 100635 (0.1 mg/kg), a selective 5-HT(1A) receptor antagonist, but it was not blocked by pretreatment with GR 127935 (0.056 mg/kg), a selective 5-HT(1B/1D) receptor antagonist. The effect of CP 94,253 (1.0 mg/kg) was blocked by pretreatment with GR 127935 (0.056 mg/kg) but was not blocked by pretreatment with WAY 100635 (0.1 mg/kg). Neither WAY 100635 nor GR 127935 altered extracellular 5-HT levels at the doses that were able to completely block the effects of either 8-OH-PIPAT or CP 94,253. The present findings suggest that, on systemic administration, both 8-OH-PIPAT and CP 94,253 are potent and selective agonists at the somatodendritic 5-HT(1A) autoreceptor and terminal 5-HT(1B/1D) autoreceptor, respectively, and are each able to cause decreases in extracellular levels of 5-HT in the mouse striatum by activating a distinct set of receptors. PMID- 10688631 TI - Quantitative prediction of metabolic inhibition of midazolam by erythromycin, diltiazem, and verapamil in rats: implication of concentrative uptake of inhibitors into liver. AB - To evaluate the degree of drug-drug interaction concerning metabolic inhibition in the liver quantitatively, we tried to predict the plasma concentration increasing ratio (R) of midazolam (MDZ) by erythromycin (EM), diltiazem (DLZ), or verapamil (VER) in rats. MDZ was administered through the portal vein at the steady state of plasma concentration of these inhibitors. The R values in the area under the plasma concentration curve of MDZ in the presence of EM, DLZ, and VER were 2.02, 1.64, and 1.30, respectively. The liver to plasma unbound concentration ratios of EM, DLZ, and VER at the steady state after infusion were 20.8, 1.02, and 3.01, respectively, suggesting concentrative uptake of EM and VER into the liver. The predicted R value in the presence of EM calculated by use of plasma unbound concentration was 1.03, whereas the value calculated with liver unbound concentration was 1.61, which was very close to the observed value. These findings indicated the need to consider the concentrative uptake of inhibitors into the liver for the quantitative prediction of metabolic inhibition. However, the predicted values in the presence of DLZ or VER calculated by use of liver unbound concentration were still underestimated. This result may be due to the metabolic inhibition by the metabolites of both inhibitors. Therefore, when predicting the degree of metabolic inhibition quantitatively, the inhibitory effect by coadministered drugs and the disposition of these metabolites in the liver must also be considered. PMID- 10688632 TI - Ligand-induced changes in surface mu-opioid receptor number: relationship to G protein activation? AB - In this study, we explored the relationship between regulation of surface mu opioid receptor number, ligand-induced G protein activation (measured by [(35)]S]guanosine-5'-O-(3-thio)triphosphate (GTPgammaS) binding) and second messenger signaling (measured by the inhibition of cAMP accumulation). Etorphine and two isomers of cis-beta-hydroxy-3-methylfentanyl (RTI-1a and RTI-1b), which were full agonists for G protein activation and signaling, caused approximately a 50% loss of surface receptors after 1 h of treatment. Fentanyl and morphine were full agonists for inhibiting cAMP accumulation and partial agonists for stimulating [(35)S]GTPgammaS binding and internalization. Although both agonists were approximately 80% as efficacious as etorphine in stimulating [(35)S]GTPgammaS binding, fentanyl induced a 35% loss of surface receptors, whereas morphine only caused a 10% loss. Additionally, both long- and short-term treatment with the opioid antagonist naloxone caused increases in surface receptors. Unexpectedly, the weak partial agonists buprenorphine and one isomer of cis-beta-hydroxy-3-methylfentanyl (RTI-1d) also were found to cause an increase in surface receptors. Treatment with pertussis toxin (PTX) diminished agonist-induced loss of surface receptors. Furthermore, the abilities of morphine and fentanyl to cause internalization were more impaired after PTX treatment than that of etorphine. PTX treatment also significantly enhanced the increase in surface receptor number caused by 18-h treatment with naloxone and buprenorphine. The results of this study suggest that disruption of G protein coupling by PTX treatment affects ligand-regulated mu-receptor trafficking and that partial agonists for signaling can vary greatly in the ability to regulate the number of surface mu-opioid receptors. PMID- 10688633 TI - Imidazoline-binding domains on monoamine oxidase B and subpopulations of enzyme. AB - A series of phenoxy-substituted methylimidazoline derivatives were synthesized and used to define the ligand recognition properties of the imidazoline-binding domain (IBD) on monoamine oxidase (MAO)-B and its role in substrate processing. The rank order of potency for selected compounds in competitive binding studies with the imidazoline [(3)H]idazoxan was different from that in enzyme activity assays, suggesting that the IBD and the site involved in enzyme inhibition are distinct. IC(50) values for inhibition of MAO-B activity by imidazoline/guanidinium ligands were one to two orders of magnitude greater than ligand concentrations that probably saturate the IBD, but were equal to the K(d) values of these ligands in competitive binding assays with the reversible MAO-B inhibitor [(3)H]Ro 19-6327. In addition, the degree of enzyme inhibition by these ligands was similar in platelet and liver, tissues exhibiting 10-fold differences in the amount of the IBD-accessible enzyme subpopulation. These data suggested that the inhibitory effect of these compounds on MAO-B activity involved a secondary interaction with the enzyme domain recognizing the inhibitor Ro 19-6327 and does not involve interaction with the IBD. Subsequent radioligand-binding studies indicated that human liver MAO-B actually existed as two distinct populations that differed in the accessibility of their IBD. The relatively small amounts of MAO-B possessing an accessible IBD ( approximately 5% in human liver) precludes determination of the functional consequences of ligand binding to the IBD. This subpopulation of MAO-B may be selectively regulated or generated in different individuals or tissues and targeted by pharmacologically active compounds in a cell type-specific manner. PMID- 10688634 TI - Kinetic and selectivity differences between rodent, rabbit, and human organic cation transporters (OCT1). AB - Organic cation transporters play an important role in the absorption, distribution, and elimination of clinical agents, toxic substances, and endogenous compounds. In kidney preparations, significant differences in functional characteristics of organic cation transport between various species have been reported. However, the underlying molecular mechanisms responsible for these interspecies differences are not known. The goal of this study was to determine the kinetics and substrate selectivities of organic cation transporter (OCT1) homologs from mouse, rat, rabbit, and human that may contribute to interspecies differences in the renal and hepatic handling of organic cations. With a series of n-tetraalkylammonium (nTAA) compounds, a correlation between increasing alkyl chain length and affinity for the four OCT1 homologs was observed. However, the apparent affinity constants (K(i)) differed among the species homologs. For the mouse homolog mOCT1, apparent K(i) values ranged from 7 microM for tetrabutylammonium to 2000 microM for tetramethylammonium. In contrast, the human homolog hOCT1 exhibited weaker interactions with the nTAA compounds. Trans-stimulation studies and current measurements in voltage-clamped oocytes demonstrated that larger nTAA compounds were transported at greater rates in oocytes expressing hOCT1, whereas smaller nTAAs were transported at greater rates in oocytes expressing mOCT1 or rOCT1. The rabbit homolog rbOCT1 exhibited intermediate properties in its interactions with nTAAs compared with its rodent and human counterparts. This report demonstrates that the human OCT1 homolog has functional properties distinct from those of the rodent and rabbit OCT1 homologs. The study underscores potential difficulties in extrapolating data from preclinical studies in animal models to humans. PMID- 10688635 TI - Cytochrome P450 omega/omega-1 hydroxylase-derived eicosanoids contribute to endothelin(A) and endothelin(B) receptor-mediated vasoconstriction to endothelin 1 in the rat preglomerular arteriole. AB - The preglomerular arteriole of the rat was used to evaluate the contribution of cytochrome P450-derived eicosanoids to the vasoconstrictor effect of endothelin (ET)-1 and to determine the receptors mediating the response. ET-1 (4 x 10(-11) to 2 x 10(-9) M) produced dose-dependent reductions in the intraluminal diameter of the renal arteriole ranging from 25 +/- 8 to 142 +/- 16 micrometer. BMS182874 [(5-dimethylamino)-N-(3, 4-dimethyl-5-isoxazolyl)-1-naphthalenesulfonamide; 3 microM], an ET(A) receptor antagonist, or BQ788 (N-cis-2, 6-dimethyl-piperidino carbonyl-L-gamma-methylleucyl-D-1-methoxy carbonyl-tryptophanyl-D-norleucine; 1 microM), an ET(B) receptor antagonist, attenuated ET-1 vasoconstriction by 59 +/- 4 and 50 +/- 10%, respectively. The combined administration of both ET receptor antagonists increased inhibition of ET-1 vasoconstriction to 75 +/- 4%. 17 Octadecynoic acid (17-ODYA, 2 microM) or 12, 12-dibromododec-enoic acid (2 microM), inhibitors of 20-hydroxyeicosatetraenoic acid (20-HETE) production, attenuated ET-1-induced vasoconstriction by 50 +/- 6 and 40 +/- 3%, respectively, as did indomethacin (10 microM), an inhibitor of cyclooxygenase. Miconazole (2 microM), the epoxygenase inhibitor, was without effect. 20-HETE (10(-8) and 2 x 10(-8) M) elicited a dose-related vasoconstriction that was inhibited by 10 microM, but not 5 microM, indomethacin. The inhibition by 17-ODYA of ET-1 vasoconstriction was not greater when combined with BMS182874 or BQ788. Moreover, vasoconstriction induced by ET-3, an ET(B)-selective agonist, was inhibited by 17 ODYA. These data indicate that both ET(A) and ET(B) receptors mediate ET-1 vasoconstriction and that 20-HETE production linked to both receptors makes a major contribution to ET-1-induced renal arteriolar vasoconstriction in the rat. PMID- 10688636 TI - Effects of selective and unselective cyclooxygenase inhibitors on prostanoid release from various rat organs. AB - It has been assumed that cyclooxygenase-2 (COX-2) is solely responsible for inflammatory processes. Recently, this view has been challenged because COX-2 selective agents caused a delay of gastric ulcer healing and exacerbation of inflammation in rats. To further characterize organ-specific toxic effects of selective and nonselective COX inhibitors, we assessed the eicosanoid release from different rat organs ex vivo after oral administration of the COX-2 selective inhibitor NS-398 and the unselective COX inhibitors diclofenac, meloxicam, and ketorolac. Prostanoid and leukotriene release from tissue fragments of the stomach, kidney, lung, and brain were determined after ex vivo incubation of tissue fragments in Tyrode's solution for 10 min at 37 degrees C. Ketorolac (0.1, 0.3, and 0.9 mg/kg) inhibited prostanoid release from all organs most potently and led to a significant increase of leukotriene release from the lung. Effects of diclofenac and meloxicam (1, 3, and 9 mg/kg each) were similar for all organs tested. At 9 mg/kg, 6keto-prostaglandin F (PGF)(1alpha) release from gastric mucosa was reduced by 79.1 +/- 11.4 and 87.6 +/- 7.7% and PGE(2) release from rat kidney was inhibited by 60.4 +/- 6.8 and 78.6 +/- 16.6% by diclofenac and meloxicam, respectively. NS-398 did not reduce prostanoid release from the lung. Consistent with the reported constitutive expression of COX-2, prostanoid release from kidney and brain was reduced by 20 to 30%. The release of 6keto-PGF(1alpha) from gastric mucosa was reduced by 34.7 +/- 22.2% at 3 mg/kg and by 86.9 +/- 12.7% at 9 mg/kg. At these doses, NS-398 has been previously shown to be COX-2 selective. Because PGF(1alpha) is the stable breakdown product of PGI(2), these results suggest that COX-2 contributes to PGI(2) synthesis in the rat stomach. PMID- 10688637 TI - Native N-methyl-D-aspartate receptors containing NR2A and NR2B subunits have pharmacologically distinct competitive antagonist binding sites. AB - The pharmacological properties of native N-methyl-D-aspartate (NMDA) receptors were determined in rat brain sections with quantitative autoradiography of [(3)H](E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid (CGP39653) binding. With five competitive antagonists as displacers, two subpopulations of binding sites were observed in the horizontal plane of section examined. These two populations corresponded anatomically to NR2A and NR2B subunits. Quantitative analysis of NR2A-like and NR2B-like binding sites was enabled by examining the cerebellar granule cell layer, which expresses NR2A and NR2C subunits, and the medial striatum, which predominately expresses NR2B subunits. The antagonists (R)-(E)-4 (3-phosphonoprop-2-enyl)piperazine-2-carboxylic acid and (R)-2-amino-5 phosphonopentanoate (D-AP5) displayed similar affinities at cerebellar NMDA receptors and medial striatal NMDA receptors. In contrast, the NMDA receptor antagonists (+/-)-6-(1H-Tetrazol-5-ylmethyl)decahydroisoquinoline- 3-carboxylic acid, (S)-alpha-amino-5-(phosphonomethyl)[1,1'-biphenyl]-3-propanoic acid, and (+/-)-cis-4-(4-phenylbenzoyl) piperazine-2,3-dicarboxylic acid displayed varied, higher affinities at medial striatal NMDA receptors than at cerebellar NMDA receptors. For the five antagonists, there was a strong correlation (r = 0.9) between the cerebellar K(i)/medial striatum K(i) ratio and the NR2A K(i)/NR2B K(i) ratio for recombinant receptors. Thus, [(3)H]CGP39653 labels two pharmacologically distinct populations of NMDA receptors that have pharmacological and anatomical properties consistent with NR2A and NR2B subunits. Because native NR2A- and NR2B-containing receptors are pharmacologically distinct, it should be possible to develop NR2A- and NR2B-selective glutamate site antagonists. PMID- 10688638 TI - ATP-dependent chromatin-remodeling complexes. PMID- 10688639 TI - BSAP can repress enhancer activity by targeting PU.1 function. AB - PU.1 and BSAP are transcription factors crucial for proper B-cell development. Absence of PU.1 results in loss of B, T, and myeloid cells, while absence of BSAP results in an early block in B-cell differentiation. Both of these proteins bind to the immunoglobulin kappa chain 3' enhancer, which is developmentally regulated during B-cell differentiation. We find here that BSAP can repress 3' enhancer activity. This repression can occur in plasmacytoma lines or in a non-B-cell line in which the enhancer is activated by addition of the appropriate enhancer binding transcription factors. We show that the transcription factor PU.1 is a target of the BSAP-mediated repression. Although PU.1 and BSAP can physically interact through their respective DNA binding domains, this interaction does not affect DNA binding. When PU.1 function is assayed in isolation on a multimerized PU.1 binding site, BSAP targets a portion of the PU.1 transactivation domain (residues 7 to 30) for repression. The BSAP inhibitory domain (residues 358 to 385) is needed for this repression. Interestingly, the coactivator protein p300 can eliminate this BSAP-mediated repression. We also show that PU.1 can inhibit BSAP transactivation and that this repression requires PU.1 amino acids 7 to 30. Transfection of p300 resulted in only a partial reversal of PU.1-mediated repression of BSAP. When PU.1 function is assayed in the context of the immunoglobulin kappa chain 3' enhancer and associated binding proteins, BSAP represses PU.1 function by a distinct mechanism. This repression does not require the PU.1 transactivation or PEST domains and cannot be reversed by p300 expression. The possible roles of BSAP and PU.1 antagonistic activities in hematopoietic development are discussed. PMID- 10688640 TI - Acetyl coenzyme A stimulates RNA polymerase II transcription and promoter binding by transcription factor IID in the absence of histones. AB - Protein acetylation has emerged as a means of controlling levels of mRNA synthesis in eukaryotic cells. Here we report that acetyl coenzyme A (acetyl-CoA) stimulates RNA polymerase II transcription in vitro in the absence of histones. The effect of acetyl-CoA on basal and activated transcription was studied in a human RNA polymerase II transcription system reconstituted from recombinant and highly purified transcription factors. Both basal and activated transcription were stimulated by the addition of acetyl-CoA to transcription reaction mixtures. By varying the concentrations of general transcription factors in the reaction mixtures, we found that acetyl-CoA decreased the concentration of TFIID required to observe transcription. Electrophoretic mobility shift assays and DNase I footprinting revealed that acetyl-CoA increased the affinity of the general transcription factor TFIID for promoter DNA in a TBP-associated factor (TAF) dependent manner. Interestingly, acetyl-CoA also caused a conformational change in the TFIID-TFIIA-promoter complex as assessed by DNase I footprinting. These results show that acetyl-CoA alters the DNA binding activity of TFIID and indicate that this biologically important cofactor functions at multiple levels to control gene expression. PMID- 10688642 TI - The Est1 subunit of yeast telomerase binds the Tlc1 telomerase RNA. AB - Est1 is a component of yeast telomerase, and est1 mutants have senescence and telomere loss phenotypes. The exact function of Est1 is not known, and it is not homologous to components of other telomerases. We previously showed that Est1 protein coimmunoprecipitates with Tlc1 (the telomerase RNA) as well as with telomerase activity. Est1 has homology to Ebs1, an uncharacterized yeast open reading frame product, including homology to a putative RNA recognition motif (RRM) of Ebs1. Deletion of EBS1 results in short telomeres. We created point mutations in a putative RRM of Est1. One mutant was unable to complement either the senescence or the telomere loss phenotype of est1 mutants. Furthermore, the mutant protein no longer coprecipitated with the Tlc1 telomerase RNA. Mutants defective in the binding of Tlc1 RNA were nevertheless capable of binding single stranded TG-rich DNA. Our data suggest that an important role of Est1 in the telomerase complex is to bind to the Tlc1 telomerase RNA via an RRM. Since Est1 can also bind telomeric DNA, Est1 may tether telomerase to the telomere. PMID- 10688641 TI - Nerve growth factor activation of the extracellular signal-regulated kinase pathway is modulated by Ca(2+) and calmodulin. AB - Nerve growth factor is a member of the neurotrophin family of trophic factors that have been reported to be essential for the survival and development of sympathetic neurons and a subset of sensory neurons. Nerve growth factor exerts its effects mainly by interaction with the specific receptor TrkA, which leads to the activation of several intracellular signaling pathways. Once activated, TrkA also allows for a rapid and moderate increase in intracellular calcium levels, which would contribute to the effects triggered by nerve growth factor in neurons. In this report, we analyzed the relationship of calcium to the activation of the Ras/extracellular signal-regulated kinase pathway in PC12 cells. We observed that calcium and calmodulin are both necessary for the acute activation of extracellular signal-regulated kinases after TrkA stimulation. We analyzed the elements of the pathway that lead to this activation, and we observed that calmodulin antagonists completely block the initial Raf-1 activation without affecting the function of upstream elements, such as Ras, Grb2, Shc, and Trk. We have broadened our study to other stimuli that activate extracellular signal-regulated kinases through tyrosine kinase receptors, and we have observed that calmodulin also modulates the activation of such kinases after epidermal growth factor receptor stimulation in PC12 cells and after TrkB stimulation in cultured chicken embryo motoneurons. Calmodulin seems to regulate the full activation of Raf-1 after Ras activation, since functional Ras is necessary for Raf-1 activation after nerve growth factor stimulation and calmodulin-Sepharose is able to precipitate Raf-1 in a calcium-dependent manner. PMID- 10688643 TI - Rap1 is a potent activation signal for leukocyte function-associated antigen 1 distinct from protein kinase C and phosphatidylinositol-3-OH kinase. AB - To identify the intracellular signals which increase the adhesiveness of leukocyte function-associated antigen 1 (LFA-1), we established an assay system for activation-dependent adhesion through LFA-1/intercellular adhesion molecule 1 ICAM-1 using mouse lymphoid cells reconstituted with human LFA-1 and then introduced constitutively active forms of signaling molecules. We found that the phorbol myristate acetate (PMA)-responsive protein kinase C (PKC) isotypes (alpha, betaI, betaII, and delta) or phosphatidylinositol-3-OH kinase (PI 3 kinase) itself activated LFA-1 to bind ICAM-1. H-Ras and Rac activated LFA-1 in a PI 3-kinase-dependent manner, whereas Rho and R-Ras had little effect. Unexpectedly, Rap1 was demonstrated to function as the most potent activator of LFA-1. Distinct from H-Ras and Rac, Rap1 increased the adhesiveness independently of PI 3-kinase, indicating that Rap1 is a novel activation signal for the integrins. Rap1 induced changes in the conformation and affinity of LFA-1 and, interestingly, caused marked LFA-1/ICAM-1-mediated cell aggregation. Furthermore, a dominant negative form of Rap1 (Rap1N17) inhibited T-cell receptor-mediated LFA 1 activation in Jurkat T cells and LFA-1/ICAM-1-dependent cell aggregation upon differentiation of HL-60 cells into macrophages, suggesting that Rap1 is critically involved in physiological processes. These unique functions of Rap1 in controlling cellular adhesion through LFA-1 suggest a pivotal role as an immunological regulator. PMID- 10688644 TI - Regulation of the resident chromosomal copy of c-myc by c-Myb is involved in myeloid leukemogenesis. AB - c-myb is a frequent target of retroviral insertional mutagenesis in murine leukemia virus-induced myeloid leukemia. Induction of the leukemogenic phenotype is generally associated with inappropriate expression of this transcriptional regulator. Despite intensive investigations, the target genes of c-myb that are specifically involved in development of these myeloid lineage neoplasms are still unknown. In vitro assays have indicated that c-myc may be a target gene of c-Myb; however, regulation of the resident chromosomal gene has not yet been demonstrated. To address this question further, we analyzed the expression of c myc in a myeloblastic cell line, M1, expressing a conditionally active c-Myb estrogen receptor fusion protein (MybER). Activation of MybER both prevented the growth arrest induced by interleukin-6 (IL-6) and rapidly restored c-myc expression in nearly terminal differentiated cells that had been exposed to IL-6 for 3 days. Restoration occurred in the presence of a protein synthesis inhibitor but not after a transcriptional block, indicating that c-myc is a direct, transcriptionally regulated target of c-Myb. c-myc is a major target that transduces Myb's proliferative signal, as shown by the ability of a c-Myc estrogen receptor fusion protein alone to also reverse growth arrest in this system. To investigate the possibility that this regulatory connection contributes to Myb's oncogenicity, we expressed a dominant negative Myb in the myeloid leukemic cell line RI-4-11. In this cell line, c-myb is activated by insertional mutagenesis and cannot be effectively down regulated by cytokine. Myb's ability to regulate c-myc's expression was also demonstrated in these cells, showing a mechanism through which the proto-oncogene c-myb can exert its oncogenic potential in myeloid lineage hematopoietic cells. PMID- 10688645 TI - An AU-rich sequence element (UUUN[A/U]U) downstream of the edited C in apolipoprotein B mRNA is a high-affinity binding site for Apobec-1: binding of Apobec-1 to this motif in the 3' untranslated region of c-myc increases mRNA stability. AB - Apobec-1, the catalytic subunit of the mammalian apolipoprotein B (apoB) mRNA editing enzyme, is a cytidine deaminase with RNA binding activity for AU-rich sequences. This RNA binding activity is required for Apobec-1 to mediate C-to-U RNA editing. Filter binding assays, using immobilized Apobec-1, demonstrate saturable binding to a 105-nt apoB RNA with a K(d) of approximately 435 nM. A series of AU-rich templates was used to identify a high-affinity ( approximately 50 nM) binding site of consensus sequence UUUN[A/U]U, with multiple copies of this sequence constituting the high-affinity binding site. In order to determine whether this consensus site could be functionally demonstrated from within an apoB RNA, circular-permutation analysis was performed, revealing one major (UUUGAU) and one minor (UU) site located 3 and 16 nucleotides, respectively, downstream of the edited base. Secondary-structure predictions reveal a stem-loop flanking the edited base with Apobec-1 binding to the consensus site(s) at an open loop. A similar consensus (AUUUA) is present in the 3' untranslated regions of several mRNAs, including that of c-myc, that are known to undergo rapid degradation. In this context, it is presumed that the consensus motif acts as a destabilizing element. As an independent test of the ability of Apobec-1 to bind to this sequence, F442A cells were transfected with Apobec-1 and the half-life of c-myc mRNA was determined following actinomycin D treatment. These studies demonstrated an increase in the half-life of c-myc mRNA from 90 to 240 min in control versus Apobec-1-expressing cells. Apobec-1 expression mutants, in which RNA binding activity is eliminated, failed to alter c-myc mRNA turnover. Taken together, the data establish a consensus binding site for Apobec-1 embedded in proximity to the edited base in apoB RNA. Binding to this site in other target RNAs raises the possibility that Apobec-1 may be involved in other aspects of RNA metabolism, independent of its role as an apoB RNA-specific cytidine deaminase. PMID- 10688646 TI - Loading of DNA-binding factors to an erythroid enhancer. AB - The HS-40 enhancer is the major cis-acting regulatory element responsible for the developmental stage- and erythroid lineage-specific expression of the human alpha like globin genes, the embryonic zeta and the adult alpha2/alpha/1. A model has been proposed in which competitive factor binding at one of the HS-40 motifs, 3' NA, modulates the capability of HS-40 to activate the embryonic zeta-globin promoter. Furthermore, this modulation was thought to be mediated through configurational changes of the HS-40 enhanceosome during development. In this study, we have further investigated the molecular basis of this model. First, human erythroid K562 cells stably integrated with various HS-40 mutants cis linked to a human alpha-globin promoter-growth hormone hybrid gene were analyzed by genomic footprinting and expression analysis. By the assay, we demonstrate that factors bound at different motifs of HS-40 indeed act in concert to build a fully functional enhanceosome. Thus, modification of factor binding at a single motif could drastically change the configuration and function of the HS-40 enhanceosome. Second, a specific 1-bp, GC-->TA mutation in the 3'-NA motif of HS 40, 3'-NA(II), has been shown previously to cause significant derepression of the embryonic zeta-globin promoter activity in erythroid cells. This derepression was hypothesized to be regulated through competitive binding of different nuclear factors, in particular AP1 and NF-E2, to the 3'-NA motif. By gel mobility shift and transient cotransfection assays, we now show that 3'-NA(II) mutation completely abolishes the binding of small MafK homodimer. Surprisingly, NF-E2 as well as AP1 can still bind to the 3'-NA(II) sequence. The association constants of both NF-E2 and AP1 are similar to their interactions with the wild-type 3'-NA motif. However, the 3'-NA(II) mutation causes an approximately twofold reduction of the binding affinity of NF-E2 factor to the 3'-NA motif. This reduction of affinity could be accounted for by a twofold-higher rate of dissociation of the NF-E2-3'-NA(II) complex. Finally, we show by chromatin immunoprecipitation experiments that only binding of NF-E2, not AP1, could be detected in vivo in K562 cells around the HS-40 region. These data exclude a role for AP1 in the developmental regulation of the human alpha-globin locus via the 3'-NA motif of HS-40 in embryonic/fetal erythroid cells. Furthermore, extrapolation of the in vitro binding studies suggests that factors other than NF-E2, such as the small Maf homodimers, are likely involved in the regulation of the HS-40 function in vivo. PMID- 10688647 TI - Recruitment of the SWI-SNF chromatin remodeling complex as a mechanism of gene activation by the glucocorticoid receptor tau1 activation domain. AB - The SWI-SNF complex has been shown to alter nucleosome conformation in an ATP dependent manner, leading to increased accessibility of nucleosomal DNA to transcription factors. In this study, we show that the SWI-SNF complex can potentiate the activity of the glucocorticoid receptor (GR) through the N terminal transactivation domain, tau1, in both yeast and mammalian cells. GR-tau1 can directly interact with purified SWI-SNF complex, and mutations in tau1 that affect the transactivation activity in vivo also directly affect tau1 interaction with SWI-SNF. Furthermore, the SWI-SNF complex can stimulate tau1-driven transcription from chromatin templates in vitro. Taken together, these results support a model in which the GR can directly recruit the SWI-SNF complex to target promoters during glucocorticoid-dependent gene activation. We also provide evidence that the SWI-SNF and SAGA complexes represent independent pathways of tau1-mediated activation but play overlapping roles that are able to compensate for one another under some conditions. PMID- 10688648 TI - Pocket protein-independent repression of urokinase-type plasminogen activator and plasminogen activator inhibitor 1 gene expression by E2F1. AB - Expression of genes of the plasminogen activator (PA) system declines at the G(0)/G(1)-S-phase boundary of the cell cycle. We found that overexpression of E2F1-3, which acts mainly in late G(1), inhibits promoter activity and endogenous expression of the urokinase-type PA (uPA) and PA inhibitor 1 (PAI-1) genes. This effect is dose dependent and conserved in evolution. Mutation analysis indicated that both the DNA-binding and transactivation domains of E2F1 are necessary for this regulation. Interestingly, an E2F1 mutant lacking the pRB-binding region strongly repressed the uPA and PAI-1 promoters. An E2F-mediated negative effect was also observed in pRB and p107/p130 knockout cell lines. This is the first report that E2F can act as a repressor independently of pocket proteins. Mutation of AP-1 elements in the uPA promoter abrogated E2F-mediated transcriptional inhibition, suggesting the involvement of AP-1 in this regulation. Results shown here identify E2F as an important component of transcriptional control of the PA system and thus provide new insights into mechanisms of cellular proliferation. PMID- 10688649 TI - The p53 tumor suppressor protein does not regulate expression of its own inhibitor, MDM2, except under conditions of stress. AB - MDM2 is an important regulator of the p53 tumor suppressor protein. MDM2 inhibits p53 by binding to it, physically blocking its ability to transactivate gene expression, and stimulating its degradation. In cultured cells, mdm2 expression can be regulated by p53. Hence, mdm2 and p53 can interact to form an autoregulatory loop in which p53 activates expression of its own inhibitor. The p53/MDM2 autoregulatory loop has been elucidated within cultured cells; however, regulation of mdm2 expression by p53 has not been demonstrated within intact tissues. Here, we examine the role of p53 in regulating mdm2 expression in vivo in order to test the hypothesis that the p53/MDM2 autoregulatory loop is the mechanism by which low levels of p53 are maintained. We demonstrate that basal expression of mdm2 in murine tissues is p53 independent, even in tissues that express functional p53. Transcription of mdm2 is induced in a p53-dependent manner following gamma irradiation, indicating that p53 regulates mdm2 expression in vivo following a stimulus. The requirement for a stimulus to activate p53 dependent regulation of mdm2 expression in vivo appeared to differ from the situation in early-passage mouse embryo fibroblasts, where mdm2 expression is enhanced by the presence of p53. Analysis of mdm2 expression in intact and dispersed embryos revealed that establishment of mouse embryo fibroblasts in culture induces p53-dependent mdm2 expression, suggesting that an unknown stimulus activates p53 function in cultured cells. Together, these results indicate that p53 does not regulate expression of its own inhibitor, except in response to stimuli. PMID- 10688650 TI - p300 requires its histone acetyltransferase activity and SRC-1 interaction domain to facilitate thyroid hormone receptor activation in chromatin. AB - We have characterized the mechanism by which coactivator p300 facilitates transcriptional activation mediated by the heterodimer of thyroid hormone (T3) receptor and 9-cis retinoid acid receptor (TR-RXR) in the context of chromatin. We demonstrate that, while p300 can enhance the transcriptional activation mediated by both liganded TR-RXR and GAL4-VP16, its histone acetyltransferase activity (HAT) is required for its ability to facilitate liganded TR-RXR- but not GAL4-VP16-mediated transcriptional activation. To understand how p300 is recruited by liganded TR-RXR, we have analyzed the interactions between TR-RXR and p300 as well as SRC-1 family coactivators. We show that, in contrast to a strong hormone-dependent interaction between TR-RXR and SRC-1 family coactivators, p300 displays minimal, if any, T3-dependent interaction with TR RXR. However, p300 can be recruited by liganded TR-RXR through its interaction with SRC-1 family coactivators. Consistent with the protein-protein interaction profile described above, we demonstrate that the SRC-1 interaction domain of p300 is important for its ability to facilitate transcriptional activation mediated by TR-RXR, whereas its nuclear receptor interaction domain is dispensable. Collectively, these results reveal the functional significance of the HAT activity of p300 and define an indirect mode for the action of p300 in TR-RXR activation. PMID- 10688651 TI - Etk, a Btk family tyrosine kinase, mediates cellular transformation by linking Src to STAT3 activation. AB - Etk (also called Bmx) is a member of the Btk tyrosine kinase family and is expressed in a variety of hematopoietic, epithelial, and endothelial cells. We have explored biological functions, regulators, and effectors of Etk. Coexpression of v-Src and Etk led to a transphosphorylation on tyrosine 566 of Etk and subsequent autophosphorylation. These events correlated with a substantial increase in the kinase activity of Etk. STAT3, which was previously shown to be activated by Etk, associated with Etk in vivo. To investigate whether Etk could mediate v-Src-induced activation of STAT3 and cell transformation, we overexpressed a dominant-negative mutant of Etk in an immortalized, untransformed rat liver epithelial cell line, WB, which contains endogenous Etk. Dominant negative inactivation of Etk not only blocked v-Src-induced tyrosine phosphorylation and activation of STAT3 but also caused a great reduction in the transforming activity of v-Src. In NIH3T3 cells, although Etk did not itself induce transformation, it effectively enhanced the transforming ability of a partially active c-Src mutant (c-Src378G). Furthermore, Etk activated STAT3 mediated gene expression in synergy with this Src mutant. Our findings thus indicate that Etk is a critical mediator of Src-induced cell transformation and STAT3 activation. The role of STAT3 in Etk-mediated transformation was also examined. Expression of Etk in a human hepatoma cell line Hep3B resulted in a significant increase in its transforming ability, and this effect was abrogated by dominant-negative inhibition of STAT3. These data strongly suggest that Etk links Src to STAT3 activation. Furthermore, Src-Etk-STAT3 is an important pathway in cellular transformation. PMID- 10688652 TI - Disassociation of met-mediated biological responses in vivo: the natural hepatocyte growth factor/scatter factor splice variant NK2 antagonizes growth but facilitates metastasis. AB - Hepatocyte growth factor/scatter factor (HGF/SF) stimulates numerous cellular activities capable of contributing to the metastatic phenotype, including growth, motility, invasiveness, and morphogenetic transformation. When inappropriately expressed in vivo, an HGF/SF transgene induces numerous hyperplastic and neoplastic lesions. NK1 and NK2 are natural splice variants of HGF/SF; all interact with a common receptor, Met. Although both agonistic and antagonistic properties have been ascribed to each isoform in vitro, NK1 retains the full spectrum of HGF/SF-like activities when expressed as a transgene in vivo. Here we report that transgenic mice broadly expressing NK2 exhibit none of the phenotypes characteristic of HGF/SF or NK1 transgenic mice. Instead, when coexpressed in NK2 HGF/SF bitransgenic mice, NK2 antagonizes the pathological consequences of HGF/SF and discourages the subcutaneous growth of transplanted Met-containing melanoma cells. Remarkably, the metastatic efficiency of these same melanoma cells is dramatically enhanced in NK2 transgenic host mice relative to wild-type recipients, rivaling levels achieved in HGF/SF and NK1 transgenic hosts. Considered in conjunction with reports that in vitro NK2 induces scatter, but not other activities, these data strongly suggest that cellular motility is a critical determinant of metastasis. Moreover, our results demonstrate how alternatively structured ligands can be exploited in vivo to functionally dissociate Met-mediated activities and their downstream pathways. PMID- 10688653 TI - RRS1, a conserved essential gene, encodes a novel regulatory protein required for ribosome biogenesis in Saccharomyces cerevisiae. AB - A secretory defect causes specific and significant transcriptional repression of both ribosomal protein and rRNA genes (K. Mizuta and J. R. Warner, Mol. Cell. Biol. 14:2493-2502, 1994), suggesting the coupling of plasma membrane and ribosome syntheses. In order to elucidate the molecular mechanism of the signaling pathway, we isolated a cold-sensitive mutant with a mutation in a gene termed RRS1 (regulator of ribosome synthesis), which appeared to be defective in the signaling pathway. The rrs1-1 mutation greatly reduced transcriptional repression of both rRNA and ribosomal protein genes that is caused by a secretory defect. RRS1 is a novel, essential gene encoding a nuclear protein of 203 amino acid residues that is conserved in eukaryotes. A conditional rrs1-null mutant was constructed by placing RRS1 under the control of the GAL1 promoter. Rrs1p depletion caused defects in processing of pre-rRNA and assembly of ribosomal subunits. PMID- 10688654 TI - The ETO protein disrupted in t(8;21)-associated acute myeloid leukemia is a corepressor for the promyelocytic leukemia zinc finger protein. AB - The ETO protein was originally identified by its fusion to the AML-1 transcription factor in translocation (8;21) associated with the M2 form of acute myeloid leukemia (AML). The resulting AML-1-ETO fusion is an aberrant transcriptional regulator due to the ability of ETO, which does not bind DNA itself, to recruit the transcriptional corepressors N-CoR, SMRT, and Sin3A and histone deacetylases. The promyelocytic leukemia zinc finger (PLZF) protein is a sequence-specific DNA-binding transcriptional factor fused to retinoic acid receptor alpha in acute promyelocytic leukemia associated with the (11;17)(q23;q21) translocation. PLZF also mediates transcriptional repression through the actions of corepressors and histone deacetylases. We found that ETO is one of the corepressors recruited by PLZF. The PLZF and ETO proteins associate in vivo and in vitro, and ETO can potentiate transcriptional repression by PLZF. The N-terminal portion of ETO forms complexes with PLZF, while the C-terminal region, which was shown to bind to N-CoR and SMRT, is required for the ability of ETO to augment transcriptional repression by PLZF. The second repression domain (RD2) of PLZF, not the POZ/BTB domain, is necessary to bind to ETO. Corepression by ETO was completely abrogated by histone deacetylase inhibitors. This identifies ETO as a cofactor for a sequence-specific transcription factor and indicates that, like other corepressors, it functions through the action of histone deactylase. PMID- 10688655 TI - ArgRII, a component of the ArgR-Mcm1 complex involved in the control of arginine metabolism in Saccharomyces cerevisiae, is the sensor of arginine. AB - Repression of arginine anabolic genes and induction of arginine catabolic genes are mediated by a three-component protein complex, interacting with specific DNA sequences in the presence of arginine. Although ArgRI and Mcm1, two MADS-box proteins, and ArgRII, a zinc cluster protein, contain putative DNA binding domains, alone they are unable to bind the arginine boxes in vitro. Using purified glutathione S-transferase fusion proteins, we demonstrate that ArgRI and ArgRII1-180 or Mcm1 and ArgRII1-180 are able to reconstitute an arginine dependent binding activity in mobility shift analysis. Binding efficiency is enhanced when the three recombinant proteins are present simultaneously. At physiological concentration, the full-length ArgRII is required to fulfill its functions; however, when ArgRII is overexpressed, the first 180 amino acids are sufficient to interact with ArgRI, Mcm1, and arginine, leading to the formation of an ArgR-Mcm1-DNA complex. Several lines of evidence indicate that ArgRII is the sensor of the effector arginine and that the binding site of arginine would be the region downstream from the zinc cluster, sharing some identity with the arginine binding domain of bacterial arginine repressors. PMID- 10688656 TI - The interaction between the Drosophila secreted protein argos and the epidermal growth factor receptor inhibits dimerization of the receptor and binding of secreted spitz to the receptor. AB - Drosophila Argos (Aos), a secreted protein with an epidermal growth factor (EGF) like domain, has been shown to inhibit the activation of the Drosophila EGF receptor (DER). However, it has not been determined whether Aos binds directly to DER or whether regulation of the DER activation occurs through some other mechanism. Using DER-expressing cells (DER/S2) and a recombinant DER extracellular domain-Fc fusion protein (DER-Fc), we have shown that Aos binds directly to the extracellular domain of DER with its carboxyl-terminal region, including the EGF-like domain. Furthermore, Aos can block the binding of secreted Spitz (sSpi), a transforming growth factor alpha-like ligand of DER, to the extracellular domain of DER. We observed that sSpi stimulates the dimerization of both the soluble DER extracellular domain (sDER) and the intact DER in the DER/S2 cells and that Aos can block the sSpi-induced dimerization of both sDER and intact DER. Moreover, we have shown that, by directly interacting with DER, Aos and SpiAos (a chimeric protein that is composed of the N-terminal region of Spi and the C-terminal region of Aos) inhibit the dimerization and phosphorylation of DER that are induced by DER's overexpression in the absence of sSpi. These results indicate that Aos exerts its inhibitory function through dual molecular mechanisms: by blocking both the receptor dimerization and the binding of activating ligand to the receptor. This is the first description of this novel inhibitory mechanism for receptor tyrosine kinases. PMID- 10688657 TI - Cloning of a mammalian transcriptional activator that binds unmethylated CpG motifs and shares a CXXC domain with DNA methyltransferase, human trithorax, and methyl-CpG binding domain protein 1. AB - Ligand screening was utilized to isolate a human cDNA that encodes a novel CpG binding protein, human CpG binding protein (hCGBP). This factor contains three cysteine-rich domains, two of which exhibit homology to the plant homeodomain finger domain. A third cysteine-rich domain conforms to the CXXC motif identified in DNA methyltransferase, human trithorax, and methyl-CpG binding domain protein 1. A fragment of hCGBP that contains the CXXC domain binds to an oligonucleotide probe containing a single CpG site, and this complex is disrupted by distinct oligonucleotide competitors that also contain a CpG motif(s). However, hCGBP fails to bind oligonucleotides in which the CpG motif is either mutated or methylated, and it does not bind to single-stranded DNA or RNA probes. Furthermore, the introduction of a CpG dinucleotide into an unrelated oligonucleotide sequence is sufficient to produce a binding site for hCGBP. Native hCGBP is detected as an 88-kDa protein by Western analysis and is ubiquitously expressed. The DNA-binding activity of native hCGBP is apparent in electrophoretic mobility shift assays, and hCGBP trans-activates promoters that contain CpG motifs but not promoters in which the CpG is ablated. These data indicate that hCGBP is a transcriptional activator that recognizes unmethylated CpG dinucleotides, suggesting a role in modulating the expression of genes located within CpG islands. PMID- 10688658 TI - Normal spermatogenesis in mice lacking the testis-specific linker histone H1t. AB - H1 histones bind to linker DNA and nucleosome core particles and facilitate the folding of chromatin into a more compact structure. Mammals contain seven nonallelic subtypes of H1, including testis-specific subtype H1t, which varies considerably in primary sequence from the other H1 subtypes. H1t is found only in pachytene spermatocytes and early, haploid spermatids, constituting as much as 55% of the linker histone associated with chromatin in these cell types. To investigate the role of H1t in spermatogenesis, we disrupted the H1t gene by homologous recombination in mouse embryonic stem cells. Mice homozygous for the mutation and completely lacking H1t protein in their germ cells were fertile and showed no detectable defect in spermatogenesis. Chromatin from H1t-deficient germ cells had a normal ratio of H1 to nucleosomes, indicating that other H1 subtypes are deposited in chromatin in place of H1t and presumably compensate for most or all H1t functions. The results indicate that despite the unique primary structure and regulated synthesis of H1t, it is not essential for proper development of mature, functional sperm. PMID- 10688659 TI - Rapid deadenylation and Poly(A)-dependent translational repression mediated by the Caenorhabditis elegans tra-2 3' untranslated region in Xenopus embryos. AB - The 3' untranslated region (3'UTR) of many eukaryotic mRNAs is essential for their control during early development. Negative translational control elements in 3'UTRs regulate pattern formation, cell fate, and sex determination in a variety of organisms. tra-2 mRNA in Caenorhabditis elegans is required for female development but must be repressed to permit spermatogenesis in hermaphrodites. Translational repression of tra-2 mRNA in C. elegans is mediated by tandemly repeated elements in its 3'UTR; these elements are called TGEs (for tra-2 and GLI element). To examine the mechanism of TGE-mediated repression, we first demonstrate that TGE-mediated translational repression occurs in Xenopus embryos and that Xenopus egg extracts contain a TGE-specific binding factor. Translational repression by the TGEs requires that the mRNA possess a poly(A) tail. We show that in C. elegans, the poly(A) tail of wild-type tra-2 mRNA is shorter than that of a mutant mRNA lacking the TGEs. To determine whether TGEs regulate poly(A) length directly, synthetic tra-2 3'UTRs with and without the TGEs were injected into Xenopus embryos. We find that TGEs accelerate the rate of deadenylation and permit the last 15 adenosines to be removed from the RNA, resulting in the accumulation of fully deadenylated molecules. We conclude that TGE-mediated translational repression involves either interference with poly(A)'s function in translation and/or regulated deadenylation. PMID- 10688660 TI - E1A-mediated repression of progesterone receptor-dependent transactivation involves inhibition of the assembly of a multisubunit coactivation complex. AB - The steroid hormone progesterone acts via high-affinity nuclear receptors that interact with specific DNA sequences located near the promoter of the hormone responsive gene. Recent studies suggested that the hormone-occupied progesterone receptor (PR) mediates gene activation by recruiting a cellular coregulatory factor, termed coactivator, to the target promoter. The identity and mechanism of action of the coactivator(s) that regulates transcriptional activity of PR are currently under investigation. Here we provide evidence that the hormone-occupied PR forms a multisubunit receptor-coactivator complex containing two previously described coactivators, CREB-binding protein (CBP) and steroid receptor coactivator 1 (SRC-1, a member of the p160 family of coactivators), in nuclear extracts of human breast tumor T47D cells. The association of CBP and SRC-1/p160 with the receptor complex is entirely hormone dependent. Both CBP and SRC-1/p160 possess intrinsic histone acetyltransferase (HAT) activity, and it has been recently proposed that these coactivators function by modulating chromatin structure at the promoter of the target gene. Interestingly, addition of purified CBP to the nuclear extracts of T47D cells markedly stimulated progesterone- and PR-dependent transcription from a nucleosome-free, progesterone response element (PRE)-linked reporter DNA template. Furthermore, depletion of SRC-1/p160 by immunoprecipitation from these transcriptional extracts also significantly impaired PR-mediated RNA synthesis from a naked PRE-linked DNA template. These results strongly implied that CBP and SRC-1/p160 facilitate receptor-mediated transcription in these cell extracts through mechanisms other than chromatin remodeling. We also observed that the adenoviral oncoprotein E1A, which interacts directly with CBP, repressed PR-mediated transactivation when added to the nuclear extracts of T47D cells. Supplementation with purified CBP overcame this inhibition, indicating that the inhibitory effect of E1A is indeed due to a blockade of CBP function. Most importantly, we noted that binding of E1A to CBP prevented the assembly of a coactivation complex containing PR, CBP, and SRC 1/p160, presumably by disrupting the interaction between CBP and SRC-1/p160. These results strongly suggested that E1A repressed receptor-mediated transcription by blocking the formation or recruitment of coactivation complexes. Collectively, our results support the hypothesis that the assembly of a multisubunit coactivation complex containing PR, CBP, and SRC-1/p160 is a critical regulatory step during hormone-dependent gene activation by PR and that the fully assembled complex has the ability to control transcription through mechanisms that are independent of the histone-modifying activities of its component coactivators. PMID- 10688661 TI - Transcriptional repression by neuron-restrictive silencer factor is mediated via the Sin3-histone deacetylase complex. AB - A large number of neuron-specific genes characterized to date are under the control of negative transcriptional regulation. Many promoter regions of neuron specific genes possess the repressor element repressor element 1/neuron restrictive silencing element (RE1/NRSE). Its cognate binding protein, REST/NRSF, is an essential transcription factor; its null mutations result in embryonic lethality, and its dominant negative mutants produce aberrant expression of neuron-specific genes. REST/NRSF acts as a regulator of neuron-specific gene expression in both nonneuronal tissue and developing neurons. Here, we shown that heterologous expression of REST/NRSF in Saccharomyces cerevisiae is able to repress transcription from yeast promoters engineered to contain RE1/NRSEs. Moreover, we have taken advantage of this observation to show that this repression requires both yeast Sin3p and Rpd3p and that REST/NRSF physically interacts with the product of the yeast SIN3 gene in vivo. Furthermore, we show that REST/NRSF binds mammalian SIN3A and HDAC-2 and requires histone deacetylase activity to repress neuronal gene transcription in both nonneuronal and neuronal cell lines. We show that REST/NRSF binding to RE1/NRSE is accompanied by a decrease in the acetylation of histones around RE1/NRSE and that this decrease requires the N-terminal Sin3p binding domain of REST/NRSF. Taken together, these data suggest that REST/NRSF represses neuronal gene transcription by recruiting the SIN3/HDAC complex. PMID- 10688662 TI - The branch point enzyme of the mevalonate pathway for protein prenylation is overexpressed in the ob/ob mouse and induced by adipogenesis. AB - We have recently reported that skeletal muscle of the ob/ob mouse, an animal model of genetic obesity with extreme insulin resistance, exhibits alterations in the expression of multiple genes. Analysis and cloning of a full-length cDNA of one of the overexpressed mRNAs revealed a 300-amino-acid protein that could be identified as the mouse geranylgeranyl diphosphate synthase (GGPP synthase) based on its homology to proteins cloned from yeast and fungus. GGPP synthase catalyzes the synthesis of all-trans-geranylgeranyl diphosphate (GGPP), an isoprenoid used for protein isoprenylation in animal cells, and is a branch point enzyme in the mevalonic acid pathway. Three mRNAs for GGPP synthase of 4.3, 3.2, and 1.7 kb were detected in Northern blot analysis. Western blot analysis of tissue homogenates using specific antipeptide antibodies revealed a single band of 34.8 kDa. Expression level of this protein in different tissues correlated with expression of the 4.3- and 3.2-kb mRNAs. GGPP synthase mRNA expression was increased 5- to 20-fold in skeletal muscle, liver, and fat of ob/ob mice by Northern blot analysis. Western blot analysis also showed a twofold overexpression of the protein in muscle and fat but not in liver, where the dominant isoform is encoded by the 1.7-kb mRNA. Differentiation of 3T3-L1 fibroblasts into adipocytes induced GGPP synthase expression more than 20-fold. Using the immunoprecipitated protein, we found that mammalian GGPP synthase synthesizes not only GGPP but also its metabolic precursor farnesyl diphosphate. Thus, the expression of GGPP synthase is regulated in multiple tissues in obesity and is induced during adipocyte differentiation. Altered regulation in the synthesis of isoprenoids for protein prenylation in obesity might be a factor determining the ability of the cells to respond to hormonal stimulation requiring both Ras-related small GTPases and trimeric G protein-coupled receptors. PMID- 10688663 TI - The H3-H4 N-terminal tail domains are the primary mediators of transcription factor IIIA access to 5S DNA within a nucleosome. AB - Reconstitution of a DNA fragment containing a Xenopus borealis somatic type 5S rRNA gene into a nucleosome greatly restricts the binding of transcription factor IIIA (TFIIIA) to its cognate DNA sequence within the internal promoter of the gene. Removal of all core histone tail domains by limited trypsin proteolysis or acetylation of the core histone tails significantly relieves this inhibition and allows TFIIIA to exhibit high-affinity binding to nucleosomal DNA. Since only a single tail or a subset of tails may be primarily responsible for this effect, we determined whether removal of the individual tail domains of the H2A-H2B dimer or the H3-H4 tetramer affects TFIIIA binding to its cognate DNA site within the 5S nucleosome in vitro. The results show that the tail domains of H3 and H4, but not those of H2A and/or H2B, directly modulate the ability of TFIIIA to bind nucleosomal DNA. In vitro transcription assays carried out with nucleosomal templates lacking individual tail domains show that transcription efficiency parallels the binding of TFIIIA. In addition, we show that the stoichiometry of core histones within the 5S DNA-core histone-TFIIIA triple complex is not changed upon TFIIIA association. Thus, TFIIIA binding occurs by displacement of H2A-H2B DNA contacts but without complete loss of the dimer from the nucleoprotein complex. These data, coupled with previous reports (M. Vettese-Dadey, P. A. Grant, T. R. Hebbes, C. Crane-Robinson, C. D. Allis, and J. L. Workman, EMBO J. 15:2508-2518, 1996; L. Howe, T. A. Ranalli, C. D. Allis, and J. Ausio, J. Biol. Chem. 273:20693-20696, 1998), suggest that the H3/H4 tails are the primary arbiters of transcription factor access to intranucleosomal DNA. PMID- 10688664 TI - Functional Cus1p is found with Hsh155p in a multiprotein splicing factor associated with U2 snRNA. AB - To explore the dynamics of snRNP structure and function, we have studied Cus1p, identified as a suppressor of U2 snRNA mutations in budding yeast. Cus1p is homologous to human SAP145, a protein present in the 17S form of the human U2 snRNP. Here, we define the Cus1p amino acids required for function in yeast. The segment of Cus1p required for binding to Hsh49p, a homolog of human SAP49, is contained within an essential region of Cus1p. Antibodies against Cus1p coimmunoprecipitate U2 snRNA, as well as Hsh155p, a protein homologous to human SAP155. Biochemical fractionation of splicing extracts and reconstitution of heat inactivated splicing extracts from strains carrying a temperature-sensitive allele of CUS1 indicate that Cus1p and Hsh155p reside in a functional, high-salt stable complex that is salt-dissociable from U2 snRNA. We propose that Cus1p, Hsh49p, and Hsh155p exist in a stable protein complex which can exchange with a core U2 snRNP and which is necessary for U2 snRNP function in prespliceosome assembly. The Cus1p complex shares functional as well as structural similarities with human SF3b. PMID- 10688665 TI - Apoptotic and growth-promoting activity of E2F modulated by MDM2. AB - E2F integrates and coordinates cell cycle progression with the transcription apparatus through its cyclical interactions with important regulators of cellular proliferation, such as pRb, cyclins, and cdk's. Physiological E2F is a heterodimeric transcription factor composed of an E2F and a DP family member, and while E2F proteins can stimulate proliferation, certain members of the family are known to be endowed with growth-inhibitory and tumor suppressor-like activity. We have investigated the product of the human mdm2 oncogene, hDM2, and report on its ability to regulate E2F-dependent apoptosis in a fashion that is independent of p53. hDM2 can prevent p53(-/-) cells from entering E2F-dependent apoptosis, an outcome that is dependent upon the presence of the DP subunit. Cells rescued from apoptosis possess lower levels of E2F subunits, although the rescued cells show an increase in DNA synthesis and possess enhanced viability that reflects cooperation between E2F-DP and hMD2. Furthermore, the regulation of E2F activity correlates with an hDM2-dependent effect on the intracellular distribution of DP 1, since hDM2 causes the nuclear accumulation of DP-1. The control of E2F by hDM2 therefore has certain parallels with the targeted degradation by MDM2 of p53. However, the domains in hDM2 required for the regulation of E2F activity can be distinguished from those necessary for p53 degradation, suggesting that control of E2F and p53 by hDM2 may be mechanistically distinct. These experiments define a new level of interplay between E2F and hDM2 whereby hDM2 has a profound impact on the physiological consequences of E2F activation. They suggest that the oncogenic properties of hDM2 may in part be mediated by an antiapoptotic activity that converts E2F from a negative to a positive regulator of cell cycle progression and thereby retains E2F at a level that contributes to a continual state of growth stimulation. PMID- 10688666 TI - Activation of apoptosis signal-regulating kinase 1 (ASK1) by tumor necrosis factor receptor-associated factor 2 requires prior dissociation of the ASK1 inhibitor thioredoxin. AB - The stress-activated protein kinases (SAPKs, also called c-Jun NH(2)-terminal kinases) and the p38s, two mitogen-activated protein kinase (MAPK) subgroups activated by cytokines of the tumor necrosis factor (TNF) family, are pivotal to the de novo gene expression elicited as part of the inflammatory response. Apoptosis signal-regulating kinase 1 (ASK1) is a MAPK kinase kinase (MAP3K) that activates both the SAPKs and p38s in vivo. Here we show that TNF receptor (TNFR) associated factor 2 (TRAF2), an adapter protein that couples TNFRs to the SAPKs and p38s, can activate ASK1 in vivo and can interact in vivo with the amino- and carboxyl-terminal noncatalytic domains of the ASK1 polypeptide. Expression of the amino-terminal noncatalytic domain of ASK1 can inhibit TNF and TRAF2 activation of SAPK. TNF can stimulate the production of reactive oxygen species (ROS), and the redox-sensing enzyme thioredoxin (Trx) is an endogenous inhibitor of ASK1. We also show that expression of TRAF2 fosters the production of ROS in transfected cells. We demonstrate that Trx significantly inhibits TRAF2 activation of SAPK and blocks the ASK1-TRAF2 interaction in a reaction reversed by oxidants. Finally, the mechanism of ASK1 activation involves, in part, homo oligomerization. We show that expression of ASK1 with TRAF2 enhances in vivo ASK1 homo-oligomerization in a manner dependent, in part, upon the TRAF2 RING effector domain and the generation of ROS. Thus, activation of ASK1 by TNF requires the ROS-mediated dissociation of Trx possibly followed by the binding of TRAF2 and consequent ASK1 homo-oligomerization. PMID- 10688667 TI - Fourteen residues of the U1 snRNP-specific U1A protein are required for homodimerization, cooperative RNA binding, and inhibition of polyadenylation. AB - It was previously shown that the human U1A protein, one of three U1 small nuclear ribonucleoprotein-specific proteins, autoregulates its own production by binding to and inhibiting the polyadenylation of its own pre-mRNA. The U1A autoregulatory complex requires two molecules of U1A protein to cooperatively bind a 50 nucleotide polyadenylation-inhibitory element (PIE) RNA located in the U1A 3' untranslated region. Based on both biochemical and nuclear magnetic resonance structural data, it was predicted that protein-protein interactions between the N terminal regions (amino acids [aa] 1 to 115) of the two U1A proteins would form the basis for cooperative binding to PIE RNA and for inhibition of polyadenylation. In this study, we not only experimentally confirmed these predictions but discovered some unexpected features of how the U1A autoregulatory complex functions. We found that the U1A protein homodimerizes in the yeast two hybrid system even when its ability to bind RNA is incapacitated. U1A dimerization requires two separate regions, both located in the N-terminal 115 residues. Using both coselection and gel mobility shift assays, U1A dimerization was also observed in vitro and found to depend on the same two regions that were found in vivo. Mutation of the second homodimerization region (aa 103 to 115) also resulted in loss of inhibition of polyadenylation and loss of cooperative binding of two U1A protein molecules to PIE RNA. This same mutation had no effect on the binding of one U1A protein molecule to PIE RNA. A peptide containing two copies of aa 103 to 115 is a potent inhibitor of polyadenylation. Based on these data, a model of the U1A autoregulatory complex is presented. PMID- 10688668 TI - Two independent signaling pathways mediate the antiapoptotic action of macrophage stimulating protein on epithelial cells. AB - In addition to its effects on macrophage function, macrophage-stimulating protein (MSP) is a growth and motility factor for epithelial cells. The growth and survival of epithelial cells generally require two signals, one generated by interaction with extracellular matrix via integrins, the other initiated by a growth factor. Therefore we investigated the effect of MSP on epithelial cell survival. Survival of epithelial cells cultured overnight in serum-free medium was promoted by adhesion, which activated both the phosphatidylinositol 3'-kinase (PI3-K)/AKT and mitogen-activated protein kinase (MAPK) pathways, operating independently of one another. The number of apoptotic cells resulting from inhibition of either pathway alone was approximately doubled by simultaneous inhibition of both pathways. This shows that each pathway made a partial contribution to the prevention of apoptosis. In the presence of an inhibitor of either pathway, MSP increased the activity of the other pathway so that the single uninhibited pathway alone was sufficient to prevent apoptosis. In contrast to the results with adherent cells, although MSP also prevented apoptosis of cells in suspension (anoikis), its effect was mediated only by the PI3-K/AKT pathway. Despite activation of MAPK by MSP, anoikis was not prevented in suspended cells with a blocked PI3-K/AKT pathway. Thus, activation of MAPK alone is not sufficient to mediate MSP antiapoptotic effects. Cell adhesion generates an additional signal, which is essential for MSP to use MAPK in an antiapoptotic pathway. This may involve translocation of MSP-activated MAPK from the cytoplasm into the nucleus, which occurs only in adherent cells. Our results suggest that there is cross talk between cell matrix adhesion and growth factors in the regulation of cell survival via the MAPK pathway. Growth factors induce MAPK activation, and adhesion mediates MAPK translocation from the cytoplasm into the nucleus. PMID- 10688669 TI - Interaction of dishevelled and Xenopus axin-related protein is required for wnt signal transduction. AB - Signaling by the Wnt family of secreted proteins plays an important role in animal development and is often misregulated in carcinogenesis. Wnt signal transduction is controlled by the rate of degradation of beta-catenin by a complex of proteins including glycogen synthase kinase 3 (GSK3), adenomatous polyposis coli, and Axin. Dishevelled is required for Wnt signal transduction, and its activation results in stabilization of beta-catenin. However, the biochemical events underlying this process remain largely unclear. Here we show that Xenopus Dishevelled (Xdsh) interacts with a Xenopus Axin-related protein (XARP). This interaction depends on the presence of the Dishevelled-Axin (DIX) domains in both XARP and Xdsh. Moreover, the same domains are essential for signal transduction through Xdsh. Finally, our data point to a possible mechanism for signal transduction, in which Xdsh prevents beta-catenin degradation by displacing GSK3 from its complex with XARP. PMID- 10688670 TI - Stimulus-specific assembly of enhancer complexes on the tumor necrosis factor alpha gene promoter. AB - The human tumor necrosis factor alpha (TNF-alpha) gene is rapidly activated in response to multiple signals of stress and inflammation. We have identified transcription factors present in the TNF-alpha enhancer complex in vivo following ionophore stimulation (ATF-2/Jun and NFAT) and virus infection (ATF-2/Jun, NFAT, and Sp1), demonstrating a novel role for NFAT and Sp1 in virus induction of gene expression. We show that virus infection results in calcium flux and calcineurin dependent NFAT dephosphorylation; however, relatively lower levels of NFAT are present in the nucleus following virus infection as compared to ionophore stimulation. Strikingly, Sp1 functionally synergizes with NFAT and ATF-2/c-jun in the activation of TNF-alpha gene transcription and selectively associates with the TNF-alpha promoter upon virus infection but not upon ionophore stimulation in vivo. We conclude that the specificity of TNF-alpha transcriptional activation is achieved through the assembly of stimulus-specific enhancer complexes and through synergistic interactions among the distinct activators within these enhancer complexes. PMID- 10688672 TI - Compensation by fibroblast growth factor 1 (FGF1) does not account for the mild phenotypic defects observed in FGF2 null mice. AB - Fibroblast growth factor 1 (FGF1) and FGF2, the prototypic members of the FGF family of growth factors, have been implicated in a variety of physiological and pathological processes. Unlike most other FGFs, FGF1 and FGF2 are ubiquitously expressed and are not efficiently secreted. Gene knockouts in mice have previously demonstrated a role for FGF2 in brain development, blood pressure regulation, and wound healing. The relatively mild phenotypic defects associated with FGF2 deletion led to the hypothesis that the continued expression of other FGFs partially compensated for the absence of FGF2 in these mice. We now report our generation of mice lacking FGF1 and their use, in combination with our previously described FGF2 null mice, to produce mice lacking both FGF1 and FGF2. FGF1-FGF2 double-knockout mice are viable and fertile and do not display any gross phenotypic defects. In the double-knockout mice we observed defects that were similar in extent to those previously described for the FGF2 null mice. Differences in the organization of neurons of the frontal motor cortex and in the rates of wound healing were observed. We also observed in FGF2(-/-) mice and in FGF1-FGF2 double-knockout mice novel impairments in hematopoiesis that were similar in severity. Essentially no abnormalities were found in mice lacking only FGF1. Our results suggest that the relatively mild defects in FGF2 knockout animals are not a consequence of compensation by FGF1 and suggest highly restricted roles for both factors under normal developmental and physiological conditions. PMID- 10688673 TI - Cytoplasmic sequestration of rel proteins by IkappaBalpha requires CRM1-dependent nuclear export. AB - Rel and IkappaB protein families form a complex cellular regulatory network. A major regulatory function of IkappaB proteins is to retain Rel proteins in the cell cytoplasm. In addition, IkappaB proteins have also been postulated to serve nuclear functions. These include the maintenance of inducible NF-kappaB-dependent gene transcription, as well as termination of inducible transcription. We show that IkappaBalpha shuttles between the nucleus and the cytoplasm, utilizing the nuclear export receptor CRM1. A CRM1-binding export sequence was identified in the N-terminal domain of IkappaBalpha but not in that of IkappaBbeta or IkappaBepsilon. By reconstituting major aspects of NF-kappaB-IkappaB sequestration in yeast, we demonstrate that cytoplasmic retention of p65 (also called RelA) by IkappaBalpha requires Crm1p-dependent nuclear export. In mammalian cells, inhibition of CRM1 by leptomycin B resulted in nuclear localization of cotransfected p65 and IkappaBalpha in COS cells and enhanced nuclear relocation of endogenous p65 in T cells. These observations suggest that the main function of IkappaBalpha is that of a nuclear export chaperone rather than a cytoplasmic tether. We propose that the nucleus is the major site of p65 IkappaBalpha association, from where these complexes must be exported in order to create the cytoplasmic pool. PMID- 10688671 TI - mSin3A regulates murine erythroleukemia cell differentiation through association with the TAL1 (or SCL) transcription factor. AB - Activation of the TAL1 (or SCL) gene is the most frequent gain-of-function mutation in T-cell acute lymphoblastic leukemia (T-ALL). TAL1 belongs to the basic helix-loop-helix (HLH) family of transcription factors that bind as heterodimers with the E2A and HEB/HTF4 gene products to a nucleotide sequence motif termed the E-box. Reported to act both as an activator and as a repressor of transcription, the mechanisms underlying TAL1-regulated gene expression are poorly understood. We report here that the corepressor mSin3A is associated with TAL1 in murine erythroleukemia (MEL) and human T-ALL cells. Interaction mapping showed that the basic-HLH domain of TAL1 was both necessary and sufficient for TAL1-mSin3A interaction. TAL1 was found, in addition, to interact with the histone deacetylase HDAC1 in vitro and in vivo, and a specific histone deacetylase inhibitor, trichostatin A (TSA), relieved TAL1-mediated repression of an E-box-containing promoter and a GAL4 reporter linked to a thymidine kinase minimal promoter. Further, TAL1 association with mSin3A and HDAC1 declined during dimethyl sulfoxide-induced differentiation of MEL cells in parallel with a decrease in mSin3A abundance. Finally, TSA had a synergistic effect with enforced TAL1 expression in stimulating MEL cells to differentiate, while constitutive expression of mSin3A inhibited MEL cell differentiation. These results demonstrate that a corepressor complex containing mSin3A and HDAC1 interacts with TAL1 and restricts its function in erythroid differentiation. This also has implications for this transcription factor's actions in leukemogenesis. PMID- 10688674 TI - Structure and function analysis of LIN-14, a temporal regulator of postembryonic developmental events in Caenorhabditis elegans. AB - During postembryonic development of Caenorhabditis elegans, the heterochronic gene lin-14 controls the timing of developmental events in diverse cell types. Three alternative lin-14 transcripts are predicted to encode isoforms of a novel nuclear protein that differ in their amino-terminal domains. In this paper, we report that the alternative amino-terminal domains of LIN-14 are dispensable and that a carboxy-terminal region within exons 9 to 13 is necessary and sufficient for in vivo LIN-14 function. A transgene capable of expressing only one of the three alternative lin-14 gene products rescues a lin-14 null mutation and is developmentally regulated by lin-4. This shows that the deployment of alternative lin-14 gene products is not critical for the ability of LIN-14 to regulate downstream genes in diverse cell types or for the in vivo regulation of LIN-14 level by lin-4. The carboxy-terminal region of LIN-14 contains an unusual expanded nuclear localization domain which is essential for LIN-14 function. These results support the view that LIN-14 controls developmental timing in C. elegans by regulating gene expression in the nucleus. PMID- 10688688 TI - Occurrence of Bacillus thuringiensis in fresh waters of Japan. AB - Bacillus thuringiensis was recovered at a relatively high frequency from both running and still fresh waters in natural environments of Kyushu, Japan. Of 107 water samples examined, 53 (49.5%) contained this organism. The frequency of B. thuringiensis colonies was 4.4% among 4414 colonies of the Bacillus cereus/B. thuringiensis group. The density of this bacterium in fresh waters averaged 0.45 cfu/ml. Serologically, B. thuringiensis isolates were assigned to 26 H serotypes. Of these, H14/36 (H serovar israelensis/malaysiensis) was the predominant, followed by the serotypes H3abc (kurstaki), H27 (mexicanensis), H3ad (sumiyoshiensis), and H35 (seoulensis). Of 195 isolates, 52 (26.7%) exhibited larvicidal activity against aquatic Diptera; 21 killed Culex pipiens molestus (Culicidae) only, and 31 were active on both the culicine mosquito and the moth fly, Clogmia albipunctata (Psychodidae). The Diptera-toxic isolates produced spherical or irregularly pointed parasporal inclusions. PMID- 10688689 TI - Effects of salt and pH stress on temperature-tolerant Rhizobium sp. NBRI330 nodulating Prosopis juliflora. AB - A study was conducted to examine the growth response of a rhizobial strain Rhizobium sp. NBRI330 isolated from root nodules of Prosopis juliflora growing in alkaline soil. The strain had the ability to nodulate P. juliflora. Nursery grown plants inoculated with Rhizobium sp. NBRI330 had 60.6% higher plant dry weight, as compared with uninoculated plants. The individual stress survival limit of a rhizobial strain Rhizobium sp. NBRI330 isolated from alkaline soil in a medium containing 32% (wt/vol) salt was 8 h, and at 55 degrees C up to 3 h. The length of Rhizobium sp. NBRI330 in salt-stressed cells increased significantly to 3.04 microm from 1.75 microm of non-stressed control cells. On the contrary, the length of pH-stressed cells declined to 1.40 microm. Compared with non-stressed control rod-shaped cells, the shape of temperature-stressed cells changed to spherical, of 0.42 microm diameter. High temperature (45 degrees C) was tolerated efficiently by Rhizobium sp. NBRI330 in the presence of salt at pH 12, as compared with pH 7. PMID- 10688690 TI - Cloning of two new cry genes from Bacillus thuringiensis subsp. wuhanensis strain. AB - With PCR products as probes, we have cloned two new cry-type genes from Bacillus thuringiensis subsp. wuhanensis. The deduced amino acid sequence of the first clone is 77.3% identical to Cry1Ga1. The deduced protein sequence of the second clone is 69.8-78.7% identical to that of Cry1B group. The nomenclature assignment of these two clones is, therefore, named Cry1Gb1 and Cry1Bd1, respectively. The Cry1Bd1 is toxic to Plutella xylostella larvae, and the Cry1Gb1 is toxic to Pieris rapae larvae. PMID- 10688691 TI - Identification of a lexA gene in, and construction of a lexA mutant of, Xanthomonas campestris pv. citri. AB - The lexA gene of Xanthomonas campestris pathovar citri (X.c. pv. citri) was cloned and sequenced. The 639-bp open reading frame encodes a protein of 213 amino acids that shares substantial sequence homology with the products of previously characterized lexA genes, sharing 46% identity with the LexA protein of Escherichia coli. Amino acids required for autocatalytic cleavage of LexA are conserved in the X.c. pv. citri protein, whereas domains thought to mediate DNA binding differ markedly from those of LexA proteins from E. coli and other bacteria. The X.c. pv. citri LexA protein was overexpressed in E. coli, and SDS polyacrylamide gel electrophoresis revealed a molecular size of 23 kDa for the purified protein. A lexA mutant of X.c. pv. citri was constructed by gene replacement, and the basal level of recA expression in this mutant was shown to be similar to that for wild-type cells exposed to a DNA-damaging agent. These results indicate that LexA functions as a repressor of recA expression in X.c. pv. citri. PMID- 10688692 TI - Purification and characterization of an extracellular alkaline serine protease from Aspergillus terreus (IJIRA 6.2). AB - An extracellular alkaline serine protease has been purified from Aspergillus terreus (IJIRA 6.2). The purification procedure involved chromatography on DEAE Sephadex A25, phosphocellulose, hydroxyapatite, casein-Sepharose, gel filtration on Sephacryl-S-300 and by glycerol density gradient centrifugation. The enzyme was further purified to apparent homogeneity through a combination of electrophoresis in polyacrylamide gel containing 0.1% sodium dodecyl sulfate (SDS) with or without protease substrate (gelatin) and subsequent regeneration of its activity in situ by removal of SDS. The active enzyme was visualized in a zymogram or on the basis of protease activity exhibited on an X-ray film. The protein in the unstained segment of the gel was electroeluted. The eluted protein with protease activity exhibited a molecular mass of 37,000-daltons on electrophoresis in SDS-polyacrylamide gel. A sedimentation coefficient of 3.2S was obtained by glycerol density gradient contrifugation. Maximum activity of protease was observed at pH 8.5 and at 37 degrees C. Purified protease was active between pH 5.5 and 9.5 and was found to be stable up to 60 degrees C. With Na caseinate, the K(m) of the purified protease was found to be 0.055 mM. Antipain, phenylmethane sulfonyl fluoride, and chymostatin served as non-competitive inhibitors. Substrate specificity was determined by using a synthetic chromogenic peptide containing N-P-Tosyl-Gly-Pro-Arg-p-nitroanilide. Results showed that the protease cleaved the peptide on the -COOH end of arginine residue. PMID- 10688693 TI - Genetic organization and polymorphism of the guaA gene encoding the GMP synthetase in Lactobacillus rhamnosus. AB - The guaA gene encoding GMP synthetase was cloned from a potential probiotic strain of Lactobacillus rhamnosus. DNA sequence and Northern blot analysis indicated that (i) guaA did not belong to an guaAB operonic structure, conversely to enteric bacteria, (ii) L. rhamnosus guaA seemed to be highly expressed, and (iii) genetic regulation might differ from Bacillus subtilis. Moreover, differences in the genetic organization of guaA allowed discrimination of some closely related L. rhamnosus strains, with a rapid screening by PCR. PMID- 10688694 TI - Secondary structural and phylogenetic implications of nuclear large subunit ribosomal RNA in the ectomycorrhizal fungus Tricholoma matsutake. AB - The sequence of large subunit (LSU) and 5.8S rRNA genes has been determined for Tricholoma matsutake. A secondary structure model was predicted for both LSU and 5.8S rRNAs, showing most of the structural features consistent with those of the consensus secondary structure model proposed for the eukaryotic cytoplasmic LSU rRNAs. With a reconstructed eukaryotic phylogeny based on full-length LSU rDNA sequences, T. matsutake was placed on the same branch with Cryptococcus neoformans as its closest neighbor. We proposed that T. matsutake be considered as one of the representative members of the division Basidiomycota. Here we report for the first time the complete LSU rRNA gene sequence in T. matsutake, a member of Homobasidiomycetes. PMID- 10688695 TI - Expression of Clostridium thermocellum endoglucanase gene in Lactobacillus gasseri and Lactobacillus johnsonii and characterization of the genetically modified probiotic lactobacilli. AB - Endoglucanase A from Clostridium thermocellum resistant to pancreatic proteinase was selected out of a range of microbial cellulases expressed in lactobacilli. Two Lactobacillus-E. coli expression vectors, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter (pSD1) and the Lactococcus lactis lac A promoter (pSD2), were constructed separately. Intestinal Lactobacillus strains, L. gasseri and L. johnsonii, were electrotransformed with pSD1 and pSD2, and the stability of each plasmid was evaluated. The endoglucanase activities of 0.722 and 0.759 U/ml were respectively found in culture medium of L. gasseri and L. johnsonii containing pSD1, and of 0.407 U/ml in medium of L. gasseri harboring pSD2. When the probiotic characteristics such as acid tolerance, bile-salt tolerance, and antibiotic susceptibility were investigated, L. gasseri and L. johnsonii were resistant to low pHs of 2 and 3. Also, L. johnsonii was bile-salt resistant in the presence of 0.5% oxgall and porcine bile extract. L. johnsonii and L. gasseri showed a rather homogeneous resistant pattern against tested antibiotics. Both strains were resistant to amikacin, bacitracin, gentamicin, streptomycin, kanamycin, and colistin. PMID- 10688696 TI - Molecular characterization of the leucine plasmid from Buchnera aphidicola, primary endosymbiont of the aphid Acyrthosiphon pisum. AB - The complete sequence of the leucine plasmid of Buchnera aphidicola from the aphid Acyrthosiphon pisum (pLeu-BAp) is reported. Its gene organization was concordant with those of other leucine plasmids of Buchnera from aphids of the Aphidini and Macrosiphini tribes. Three inverted repeats are present in pLeu-BAp. Two of them are also present in pLeu from the family Aphididae: (i) SIR1, located downstream the leucine operon, resembles a rho-independent terminator of transcription, and (ii) LIR1, located upstream of the leucine operon, is suggested to be involved in transcription termination or messenger stability. The third, located near the putative ATGC repeats involved in the origin of replication, is specific in aphids of the Macrosiphini tribe. Phylogenetic analyses based on sequences of leuA, leuB, leuC, leuD, repA1 and ORF1 showed a closer relationship between Buchnera (A. pisum) and Buchnera (Diuraphis noxia). However, tree topologies indicate that the split between both aphid species took place soon after the formation of the Macrosiphini lineage. PMID- 10688697 TI - Uniparental mitochondrial transmission in sexual crosses in Cryptococcus neoformans. AB - Restriction fragment length polymorphism (RFLP) in the large ribosomal RNA region of the mitochondrial DNA (mtDNA) was developed as a genetic marker for investigating mitochondrial transmission in sexual crosses of the human pathogenic basidiomycetous yeast Cryptococcus neoformans. Strain JEC20 of C. neoformans var. neoformans (mat a) was mated with six strains of C. neoformans var. grubii (mat alpha). Successful mating was indicated by the formation of hyphae and basidiospores. These basidiospores were examined for mtDNA RFLP genotypes. All 570 basidiospores examined from the six crosses showed the mtDNA genotype of strain JEC20. The failure to recover the C. neoformans var. grubii mtDNA in any cross indicates that the C. neoformans var. grubii mtDNA is either selectively eliminated in the newly formed dikaryon or selectively excluded in the immediate dikaryotic hyphae of the newly formed dikaryon. PMID- 10688698 TI - Characterization of the extracellular polysaccharide produced by a marine cyanobacterium, Cyanothece sp. ATCC 51142, and its exploitation toward metal removal from solutions. AB - Cyanobacterium, Cyanothece sp. ATCC 51142 produces an exopolysaccharide at a high level. Physical analysis of the exopolysaccharide (EPS), such as nuclear magnetic resonance, infrared spectrum, were done to determine its possible structure. Thermal gravimetric analysis, differential scanning calorimeter, and differential thermal analysis of the polymer were done to find out the thermal behavior. Calcium content within the sample was found out. Some of the physicochemical properties, such as relative viscosity, specific viscosity, and intrinsic viscosity of the EPS were studied under different conditions. The phenomenon of gel formation by the EPS was investigated for its potential application in metal removal from solutions. PMID- 10688699 TI - Strains of Xylella fastidiosa rapidly distinguished by arbitrarily primed-PCR. AB - Genomic DNAs isolated from strains of Xylella fastidiosa that caused citrus variegated chlorosis, coffee leaf scorch, Pierce's Disease of grapevine, and plum leaf scorch were analyzed by arbitrarily primed polymerase chain reaction. Purified DNA was amplified under nonstringent conditions with single primers 21 nucleotides (nt) long. Thirty-nine amplification products were observed that were useful to distinguish among the strains and to derive a similarity matrix and construct a phenogram showing possible relationships among the strains. Strains isolated from diseased coffee and citrus in Brazil were closely related to each other (coefficient of similarity of 0. 872), but only distantly related to a strain isolated from diseased grapevine in the USA (coefficient of similarity of 0.650). Strains of Xylella fastidiosa isolated from diseased plums in the USA and Brazil clustered with strains from different hosts isolated from their respective countries of origin. Thus, there may be two quite dissimilar clusters of strains of Xylella fastidiosa, one in North America and the other in South America. Each cluster contains strains that can cause disease in plum. The methods described provide a convenient and rapid method to distinguish between strains of Xylella fastidiosa that cause diseases of coffee and citrus in the same region of Brazil. This has not been possible previously. This will potentially enable the two strains to be distinguished in alternate hosts or in insect vectors. PMID- 10688700 TI - HSP16.6 is involved in the development of thermotolerance and thylakoid stability in the unicellular cyanobacterium, Synechocystis sp. PCC 6803. AB - The low molecular weight (LMW) heat shock protein (HSP), HSP16.6, in the unicellular cyanobacterium, Synechocystis sp. PCC 6803, protects cells from elevated temperatures. A 95% reduction in the survival of mutant cells with an inactivated hsp16.6 was observed after exposure for 1 h at 47 degrees C. Wild type cell survival was reduced to only 41%. HSP16.6 is also involved in the development of thermotolerance. After a sublethal heat shock at 43 degrees C for 1 h and subsequent challenge exposure at 49 degrees C for 40 min, mutant cells did not survive, while 64% of wild-type cells survived. Ultrastructural changes in the integrity of thylakoid membranes of heat-shocked mutant cells also are discussed. These results demonstrate an important protective role for HSP16.6 in the protection of cells and, in particular, thylakoid membrane against thermal stress. PMID- 10688702 TI - Routes of Ethephon Uptake in Pineapple (Ananas comosus) and Reasons for Failure of Flower Induction. AB - Ethylene-releasing agents such as ethephon (2-chloroethylphosphonic acid) are used widely to induce flowering in pineapples (Ananas comosus (L.) Merrill). However, ethephon treatment is less reliable in summer, particularly if plants are treated on abnormally hot days. [(14)C]ethephon was used to follow uptake and translocation in leaf tissues. Up to 30% of the ethephon entered the leaf within 4 h, and up to 60% by 24 h. Uptake was dramatically modified by temperature, relative humidity, solution pH, and the surface on which solution droplets were placed. Entry occurred across the leaf cuticle and probably also by way of stomatal pores, and label was recovered at all depths within the leaf. (14)C label entered more rapidly through the abaxial epidermis than through the adaxial epidermis. Low-volume spray applications to whole plants resulted in rapidly drying droplets mainly on the adaxial, distal epidermis and were rather ineffective at inducing flowering, possibly because little ethephon or ethylene reaches the shoot apex. High-volume sprays may facilitate ethephon entry because solution accumulates in leaf axils and hence remains in prolonged contact with abaxial epidermis of leaf bases close to the shoot apex. When poured into the center of the plant, 20% of a normal commercial ethephon dose induced full flowering even under adverse temperatures. It is suggested that high-volume evening spraying and avoidance of hot days may reduce the incidence of flowering failure. PMID- 10688703 TI - Opening of Rice Floret in Rapid Response to Methyl Jasmonate. AB - Effects of methyl jasmonate (MeJA) on rice floret opening were investigated in seven cultivars or hybrid combinations covering various variety types. Intact or excised panicles, judged to have florets just before anthesis, were soaked in 4 x 10(-5) - 4 x 10(-3)M MeJA solutions for 2 min at different temperatures. The results indicated that MeJA significantly induced opening of rice florets within about 30 min, with the most rapid induction occurring just 6 min after treatment. Numbers of induced opening florets are correlated with MeJA concentrations. Higher concentrations of MeJA induced more florets. pH values had no influence on MeJA effect, but MeJA required less time and induced more florets at 34 degrees C than at 25 degrees C. As far as we know, this is the first evidence that floret opening is induced by plant hormones. CO(2) evolution from panicles was also increased by MeJA treatment. Field experiments revealed that perfect flowering synchrony between the cytoplasmic male sterile (CMS) and restorer lines in hybrid seed production could be obtained by spraying MeJA solution on CMS line plants at the rate of 25 mg/m(2). As a result, many more hybrid seeds were harvested. PMID- 10688704 TI - Putrescine Influences Growth and Production of Coumarins in Hairy Root Cultures of Witloof Chicory (Cichorium intybus L. cv. Lucknow Local). AB - The effect of putrescine (Put) on the growth and production of two coumarins, esculin and esculetin, in hairy roots of chicory (Cichorium intybus L. cv. Lucknow local) was examined. To study the role of Put on growth and production of coumarins, polyamine inhibitors, namely alpha-dl-difluromethylornithine and alpha dl-difluromethylarginine were used at 1 mM concentration. Put treatment at 1.5 mM produced a 1.9-fold increase in the growth of hairy roots, as well as the production of esculin and esculetin. The treatments with polyamine (PA) inhibitors resulted in much lower growth and production of coumarins compared with both 1.5-mM Put treatment and the control. Both free and conjugated PAs were studied over the whole culture period, and conjugates of all three PAs, namely Put, spermidine, and spermine, were higher than free PAs throughout the culture period. The treatments with PA inhibitors showed lower levels of endogenous PAs compared with Put-treated samples. The treatment with 1.5 mM Put showed maximum accumulation of endogenous conjugated Put (2,098 +/- 157 nmoles gm(-1) fresh weight). The production of esculin and esculetin was strictly correlated with growth in all treatments. Put at 1.5 mM resulted in greater length of primary root (18.3 +/- 1.4 cm) as compared with the control (11 +/- 0.9 cm) and larger numbers of secondary and tertiary roots. PMID- 10688705 TI - Gibberellins and Subapical Cell Divisions in Relation to Bud Set and Bud Break in Salix pentandra. AB - In young plants of Salix pentandra, a temperate zone deciduous woody species, elongation growth ceases and a terminal bud is formed at day lengths shorter than a critical length. This is the first step in dormancy development, making survival under harsh winter conditions possible. Early studies strongly indicate that gibberellin is involved in the photoperiodic control of bud set and bud break. GA(1) action was studied by application under short days to plants where cessation of shoot elongation had occurred, followed by subsequent anatomic investigations of shoot tips. Under short days the frequency of cell division decreased rapidly along with the earlier observed decrease in GA(1) levels. Application of GA(1) to short-day-induced terminal buds rapidly stimulated cell division in apices several days before visible shoot elongation in response to this treatment was observed. One day after GA(1) application a fourfold increase in cell division frequency in apices was observed, increasing to a maximum of sevenfold 2 days after application. Long-day treatment leading to induction of bud break after about 4-6 days was followed by slowly increasing frequency of cell divisions. In earlier studies of this species, short days and gibberellins had no effect on cell elongation. These data show that increased GA(1) content, by application or long-day treatment, results in increased frequency of mitosis. This strongly indicates that GA(1) affects stem elongation in connection with bud set and bud break primarily by affecting cell divisions in subapical tissues. PMID- 10688706 TI - Herbicidal Derivatives of Aminomethylenebisphosphonic Acid. Part IV. Hydroxyalkylidenebisphosphonates, Iminomethylenebisphosphonates and Ureidomethylenebisphosphonates. AB - Derivatives of aminomethylenebisphosphonic acids constitute a class of promising herbicides. Replacement of the amino group by hydroxyl, ureido, thioureido, or imino moieties leads to compounds of significant herbicidal properties. This indicates that protonated amino function is not a requirement for phytotoxic action of bisphosphonates. PMID- 10688707 TI - New Plant Growth Regulators Protect Photosynthesis and Enhance Growth Under Drought of Jack Pine Seedlings. AB - To determine whether natural plant growth regulators (PGRs) can enhance drought tolerance and the competitive ability of transplanted seedlings, 1.5-year-old jack pine (Pinus banksana Lamb.) seedlings were treated with homobrassinolide, salicylic acid, and two polyamines, spermine and spermidine, triacontanol, abscisic acid (ABA), and the synthetic antioxidant, Ambiol. PGRs were fed into the xylem for 7 days and plants were droughted by withholding water for 12 days. ABA, Ambiol, spermidine, and spermine at a concentration of 10 ug L(-1) stimulated elongation growth under drought, whereas ABA, Ambiol, and spermidine maintained higher photosynthetic rates, higher water use efficiency, and lower Ci/Ca ratio under drought compared with control plants. The damaging effects of drought on membrane leakage was reversed by Ambiol, ABA, triacontanol, spermidine, and spermine. Because ABA, Ambiol, and both polyamines enhanced elongation growth and also reduced membrane damage in jack pine under drought, they show promise as treatments to harden seedlings against environmental stress. The protective action of these compounds on membrane integrity was associated with an inhibition of ethylene evolution, with a reduction in transpiration rate and an enhancement of photosynthesis, which together increased water use efficiency under drought. Although most of the tested compounds acted as antitranspirants, the inhibition in membrane leakage in ABA-, Ambiol-, and polyamine-treated plants appeared more closely related to the antiethylene action. PMID- 10688708 TI - Influence of 2,3,5-Triiodobenzoic Acid and 1-N-Naphthylphthalamic Acid on Indoleacetic Acid Transport in Carnation Cuttings: Relationship with Rooting. AB - (3)H-IAA transport in excised sections of carnation cuttings was studied by using two receiver systems for recovery of transported radioactivity: agar blocks (A) and wells containing a buffer solution (B). When receivers were periodically renewed, transport continued for up to 8 h and ceased before 24 h. If receivers were not renewed, IAA transport decreased drastically due to immobilization in the base of the sections. TIBA was as effective as NPA in inhibiting the basipetal transport irrespective of the application site (the basal or the apical side of sections). The polarity of IAA transport was determined by measuring the polar ratio (basipetal/acropetal) and the inhibition caused by TIBA or NPA. The polar ratio varied with receiver, whereas the inhibition by TIBA or NPA was similar. Distribution of immobilized radioactivity along the sections after a transport period of 24 h showed that the application of TIBA to the apical side or NPA to the basal side of sections, increased the radioactivity in zones further from the application site, which agrees with a basipetal and acropetal movement of TIBA and NPA, respectively. The existence of a slow acropetal movement of the inhibitor was confirmed by using (3)H-NPA. From the results obtained, a methodological approach is proposed to measure the variations in polar auxin transport. This method was used to investigate whether the variations in rooting observed during the cold storage of cuttings might be related to changes in polar auxin transport. As the storage period increased, a decrease in intensity and polarity of auxin transport occurred, which was accompanied by a delay in the formation and growth of adventitious roots, confirming the involvement of polar auxin transport in supplying the auxin for rooting. PMID- 10688709 TI - Photodynamic therapy in dermatology. AB - The combination of light and chemicals to treat skin diseases is widely practiced in dermatology. Within this broad use of light and drugs, in recent years the concept of photodynamic therapy (PDT) has emerged. PDT is a promising modality for the management of various tumors and nonmalignant diseases, based on the combination of a photosensitizer that is selectively localized in the target tissue and illumination of the lesion with visible light, resulting in photodamage and subsequent cell death. Moreover, the fluorescence of photosensitizing compounds is also utilized as a helpful diagnostic tool for the detection of neoplastic tissue. Intensive basic and clinical research culminated in the worldwide approval of PDT for bladder, esophageal, and lung cancer. The expanding use of this relatively new therapeutic modality in dermatology at many centers around the world has revealed its efficacy for the treatment of cutaneous precancer and cancer, as well as selected benign skin disorders. The following article summarizes the main principles of PDT considering the most recent developments and provides a comprehensive synopsis of the present status of the use of PDT in dermatology. (J Am Acad Dermatol 2000;42:389-413.) LEARNING OBJECTIVE: At the conclusion of this learning activity, participants should be able to describe the basic concepts of PDT, including fundamental knowledge of the most relevant photosensitizers, the light sources, the mechanisms involved in PDT-mediated cell destruction, as well as the indications and limitations of photodynamic treatment of skin diseases. PMID- 10688710 TI - Patch granuloma annulare: clinicopathologic study of 6 patients. AB - BACKGROUND: Granuloma annulare is a common skin disorder that usually presents with smooth papules arranged as annular plaques. Variants, such as disseminated, subcutaneous, and perforating granuloma annulare, have been described. OBJECTIVE: The purpose of this study is to describe the clinical and histologic features of a distinct patch form of granuloma annulare. METHODS: The clinical and histologic features of 6 patients with patch granuloma annulare were evaluated. RESULTS: Six women 27 to 72 years of age had patches on the extremities. Two patients also had a lesion on the trunk. Only one patient had annular patches. Histologic examination showed an interstitial infiltrate of lymphocytes and histiocytes with diffuse necrobiosis. CONCLUSION: Patch granuloma annulare is a distinct variant with rather subtle clinical and histologic features. A high index of suspicion both clinically and histologically aids in making the diagnosis. PMID- 10688711 TI - Patterns of remission in pemphigus vulgaris. AB - BACKGROUND: The incidence of remissions in pemphigus is unclear because these are usually reported at a single point in the evolution of the disease. Thus it is uncertain whether treatment simply suppresses the manifestations of the disease and consequently must be continuously administered, or induces complete and long lasting remissions that permit therapy to be discontinued. OBJECTIVE: To answer this question, we investigated the incidence of remission in a long-term longitudinal study. METHODS: The induction of complete and long-lasting remissions (lesion free with no systemic therapy for at least 6 months) was studied in 40 patients with pemphigus vulgaris treated conventionally and followed up for an average of 7.7 years by the same investigator. RESULTS: Five (5%) of the patients died of the disease. Complete and long-lasting remissions were induced in 25%, 50%, and 75% of patients 2, 5, and 10 years, respectively, after diagnosis. Most of the remaining patients were in partial remission or had mild disease controlled with a small dose of steroids. The course of the disease followed different patterns, with some patients rapidly entering complete and long-lasting remissions, whereas others never entered into a complete remission. The induction of complete remission was related to the initial severity and extent of disease and to early response to treatment. CONCLUSION: It is possible to eventually induce complete and durable remissions in most patients with pemphigus that permit systemic therapy to be safely discontinued without a flare in disease activity. The proportion of patients in whom this can be achieved increases steadily with time, and therapy can be discontinued in approximately 75% of patients after 10 years. PMID- 10688712 TI - Effects of administration of a single dose of a humanized monoclonal antibody to CD11a on the immunobiology and clinical activity of psoriasis. AB - BACKGROUND: CD11a/CD18 comprise subunits of leukocyte function associated antigen (LFA-1), a T-cell surface molecule important in T-cell activation, T-cell emigration into skin, and cytotoxic T-cell function. OBJECTIVE: We explored the immunobiologic and clinical effects of treating moderate to severe psoriasis vulgaris with a single dose of humanized monoclonal antibody against CD11a (hu1124). METHODS: This was an open label study with a single dose of hu1124 at doses of 0.03 to 10 mg/kg. Clinical (Psoriasis Area and Severity Index [PASI]) and immunohistologic parameters (epidermal thickness, epidermal and dermal T-cell numbers, and keratinocyte intercellular adhesion molecule 1 [ICAM-1] expression) were followed. RESULTS: Treatment with hu1124, at doses higher than 1.0 mg/kg (group III), completely blocks CD11a staining for at least 14 days in both blood and psoriatic plaques. At 0.3 to 1.0 mg/kg, T-cell CD11a staining was completely blocked; however, blockade lasted less than 2 weeks (group II). Only partial saturation of either blood or plaque cellular CD11a was observed at doses of hu1124 between 0.01 and 0.1 mg/kg (group I). This pharmacodynamic response was accompanied by decreased numbers of epidermal and dermal CD3(+) T cells, decreased keratinocyte and blood vessel expression of ICAM-1, and epidermal thinning. Statistically significant drops in PASI compared with baseline were observed in group II patients at weeks 3 and 4 and in group III patients at weeks 2 through 10. No significant drop in PASI score was observed in group 1. Adverse events were mild at doses of 0.3 mg/kg or less and included mild chills, abdominal discomfort, headache, and fever. At a single dose of 0.6 mg/kg or higher, headache was the most common dose-limiting toxicity observed. CONCLUSION: Targeting CD11a may improve psoriasis by inhibiting T-cell activation, T-cell emigration into the skin, and cytotoxic T-cell function. PMID- 10688713 TI - Rate of body dysmorphic disorder in dermatology patients. AB - BACKGROUND: Dermatologists appear to be the physicians most often seen by patients with the psychiatric condition body dysmorphic disorder (BDD), a distressing or impairing preoccupation with a nonexistent or slight defect in appearance. The frequency of BDD among patients seeking dermatologic treatment is unknown, however. OBJECTIVE: This study determined the percentage of patients seeking dermatologic treatment who screened positive for BDD. METHODS: A validated self-report questionnaire and a reliable defect severity scale were used to determine the rate of BDD in 268 patients seeking dermatologic treatment. RESULTS: A total of 11.9% (95% confidence interval [CI], 8.0%-15.8%) of patients screened positive for BDD. Rates were similar in a community general dermatology setting (14.4% [95% CI, 8.5%-20.3%]) and a university cosmetic surgery setting (10. 0% [95% CI, 6.1%-13.9%]). CONCLUSION: BDD appears relatively common among patients seeking dermatologic treatment. Further research is needed to confirm these findings and to assist dermatologists in identifying these potentially high risk patients. PMID- 10688714 TI - Davener's dermatosis: a variant of friction hypermelanosis. AB - BACKGROUND: As part of our clinical experience we encountered a group of patients from a specific population with a similar peculiar pigmentation over the lower dorsal spine. OBJECTIVE: We investigated these patients to see whether we could determine a common origin. METHODS: Patients meeting the inclusion criteria underwent detailed history and complete physical examination; biopsy specimens from 3 patients were studied. RESULTS: All 13 patients were full-time male students at Orthodox Jewish Talmudic seminaries (Yeshivas). The lesion consisted of an elongated, vertical, midline, hyperpigmented patch with indistinct borders, which was distributed along the skin overlying the bony protuberances of the inferior thoracic and lumbar vertebrae. It was often unrecognized by the patients. Mean body mass index was lower than that for the general population. Histologic study showed a marked diffuse hyperkeratosis and hyperplastic epidermis with diffuse hyperpigmentation. We attributed the phenomenon to friction from the rigid backrests against the cutaneous surface of the lower back generated by the characteristic swaying activity that traditionally accompanies Torah study or "davening" (praying) and termed it Davener's dermatosis. CONCLUSION: We believe this phenomenon represents a new form of benign friction hypermelanosis. This report highlights the importance of a thorough history in patients presenting with pigmented lesions. PMID- 10688715 TI - Host-related and environmental risk factors for cutaneous basal cell carcinoma: evidence from an Italian case-control study. AB - BACKGROUND: Despite its frequency, there is a paucity of data on risk factors for basal cell carcinoma. OBJECTIVE: We assessed potential risk factors for basal cell carcinoma in a population from southern Europe. METHODS: This multicenter case-control study involved 528 newly diagnosed cases and 512 controls. RESULTS: In the multivariate analysis, red hair, lighter colored eyes, high nevus counts on the upper limbs, and the presence of solar lentigines and actinic keratoses were all associated with basal cell carcinoma. The risk of the tumor increased in subjects who reported burning easily and experiencing sunburn episodes before 15 years of age. An association was documented with indices of recreational sun exposure but no clear evidence of exposure-effect relationship was found. No relation was found with occupational sun exposure. Finally, basal cell carcinoma appeared to be significantly associated with a family history of skin tumors, a personal history of tumors other than those on skin, and radiotherapy. CONCLUSION: Genetic and environmental factors appear to be involved in the onset of basal cell carcinoma. PMID- 10688716 TI - The radiation accident in Georgia: clinical appearance and diagnosis of cutaneous radiation syndrome. AB - BACKGROUND: Eleven male Georgian soldiers were accidentally exposed to radiation by cesium 137 during their training in a military exercise camp in Lilo, Georgia between November 1996 and May 1997. OBJECTIVE: The characteristic sequelae of accidental cutaneous irradiation and available diagnostic methods are described. METHODS: Magnetic resonance imaging (MRI) of radiation ulcers was performed in all patients; thermography was performed in 2. In 7 patients ulcers and white macules were examined with high-frequency 20 MHz sonography; histologic results were obtained from all patients. RESULTS: Predominant lesions were radiation ulcers in 11 patients and white hairless macules in 7. MRI showed ulcers down to the muscles and an increase of signal intensity in the musculature in 9 cases. The corresponding muscle histology demonstrated vasculitis in 7 patients and necrosis in 2. In 2 patients, MRI signal intensity of the musculature was normal. In 3 patients, 20 MHz sonography showed dermal defects; 1 patient had cutaneous fibrosis. Thermography demonstrated hypothermic zones with extended inflammatory zones adjacent to the radiation ulcers in both patients examined. CONCLUSION: High-frequency 20 MHz sonography, MRI, and thermography are useful noninvasive methods for diagnosis of the extent of cutaneous radiation syndrome and for therapy planning. PMID- 10688717 TI - The combination of 2% 4-hydroxyanisole (Mequinol) and 0.01% tretinoin is effective in improving the appearance of solar lentigines and related hyperpigmented lesions in two double-blind multicenter clinical studies. AB - BACKGROUND: Solar lentigines are a chronic condition of the aging population resulting from years of cumulative sun exposure. A topical treatment that is both safe and effective would be welcome and useful. Combinations of therapeutic agents are often used and allow synergy of mechanisms with tolerability. A tyrosinase inhibitor in use in Europe, 4-hydroxyanisole (Mequinol), and the retinoid tretinoin have been used singly as depigmenting agents. OBJECTIVE: The efficacy and safety of the combination product of 2% 4-hydroxyanisole (4HA [mequinol]) /0.01% tretinoin solution (tradename Solage) were evaluated in two phase III, randomized, controlled, double-blind trials. METHODS: Subjects were randomized to treatment with 4HA/tretinoin solution, one of the active components (4HA or tretinoin), or vehicle. Subjects applied the test solution with a wand applicator twice daily to all solar lentigines and related hyperpigmented lesions on the face, forearms, and backs of hands for up to 24 weeks. Trial 1 had a 24 week no-treatment regression phase and trial 2 had a 4-week no-treatment regression phase. Information collected included clinical assessments of Target Lesion Pigmentation, Physician's Global Assessment of Improvement/Worsening, an Assessment of Overall Cosmetic Effect, and a Subject's Self-Assessment Questionnaire. RESULTS: The 4HA/tretinoin combination was clinically superior to each of its active components and to the vehicle in the treatment of solar lentigines. At the end of treatment, in trial 1 and trial 2, 4HA/tretinoin was statistically superior to each of its active components and vehicle on the forearms and face (P 1000 J/cm(2)) through PUVA therapy. All patients had PUVA-induced keratoses elsewhere, but the number varied greatly between patients. The hand and foot keratoses were well defined and circular and had a characteristic histologic appearance, with a sharp demarcation between normal and abnormal markedly pale-staining epidermis. CONCLUSION: These lesions are a further cutaneous manifestation of prolonged PUVA therapy. PMID- 10688721 TI - Newer strategies for effective evaluation of primary melanoma and treatment of stage III and IV disease. AB - Our objective in this article is to update dermatologists on the clinical management of malignant melanoma, including the role of sentinel node biopsy and options for the treatment of stage III and stage IV disease. The role of the dermatologist throughout the continuum of care is emphasized, and the essential partnership with medical oncology in recognizing promising new options for patients with advanced disease is examined. PMID- 10688722 TI - Potassium iodide and the wolff-chaikoff effect: relevance for the dermatologist. AB - The inhibition of organic binding of iodide in the thyroid gland by excess iodide, resulting in the cessation of thyroid hormone synthesis, is known as the Wolff-Chaikoff effect. This review explores the nature of the Wolff-Chaikoff effect, both in terms of its potential mechanisms and its relevance to dermatologists who use potassium iodide as a therapeutic agent. PMID- 10688723 TI - Combination phototherapy of psoriasis with narrow-band UVB irradiation and topical tazarotene gel. AB - BACKGROUND: Narrow-band UVB (311 nm) phototherapy offering an emission spectrum closely conforming to the peak of the action spectrum for clearing psoriasis has significantly improved phototherapy for psoriasis. Because the majority of the commonly used topical therapies in treatment of psoriasis have limitations, a need for new topical agents remains. Tazarotene has been shown to be efficacious in plaque-type psoriasis. Combination of narrow-band UVB with topical agents has been shown to enhance efficacy of both treatment modalities. OBJECTIVE: We attempted to evaluate the efficacy of narrow-band UVB phototherapy in combination with topical tazarotene. METHODS: Ten patients with stable plaque psoriasis were treated with narrow-band UVB. In addition, topical tazarotene 0.05% was applied once daily to one side of the body. The follow-up period was 4 weeks. Efficacy was assessed separately for both body halves by means of a modified Psoriasis Area and Severity Index (PASI). RESULTS: Both treatment modalities notably reduced the PASI scores with values being significantly lower in skin areas treated with narrow-band UVB phototherapy in combination with topical tazarotene. CONCLUSION: The addition of tazarotene to narrow-band UVB phototherapy promotes more effective, faster clearing of psoriasis compared with UVB (311 nm) monotherapy. PMID- 10688724 TI - A comparative immunohistochemical study of MART-1 expression in Spitz nevi, ordinary melanocytic nevi, and malignant melanomas. AB - BACKGROUND: The histopathologic differential diagnosis of Spitz nevus (SN) from malignant melanoma (MM) may be difficult. OBJECTIVE: We attempted to elucidate the pattern of expression of a newly recognized melanocyte-specific melanosomal protein MART-1 in routinely processed specimens of SNs, MMs, and ordinary melanocytic nevi (MNs) and to see whether it can help to differentiate between them. METHODS: Twenty SN, 22 MM, and 27 ordinary MN were immunostained with anti MART-1 monoclonal antibody (clone A103). RESULTS: All SNs, MNs, and MMs demonstrated cytoplasmic staining for MART-1 in some of their tumor cells, of which 17 of 20 (85%) and 24 of 27 (89%) of SN and MN, respectively, demonstrated positive stainings in more than half of their tumor cells, as compared with only 10 of 22 (45%) of the MM (P <.05). The majority of lesions in all 3 types of tumors showed a homogeneous mode of staining, although MM tended to show a more heterogeneous pattern. A consistent pattern of stratification of staining with progressive descent into the dermis was not demonstrated in these tumors. CONCLUSION: MART-1 does not differentiate between SN, MM, and ordinary MN in a consistent pattern, but it may be used as a marker for these tumors. PMID- 10688725 TI - Skin disorders in amputees. AB - BACKGROUND: Dermatologic problems restrict the normal use of a prosthetic limb. The importance of contact dermatitis to skin morbidity in a population of amputees and the selection criteria for patch testing have not been clearly defined. OBJECTIVE: We describe the range of dermatoses seen in a population of amputees and examine the incidence, causes, and patterns of contact dermatitis. METHODS: This is a questionnaire-based, cross-sectional study of 210 amputees. Those with a skin problem were assessed by a dermatologist. Patch testing was undertaken in patients with persistent dermatitis. RESULTS: A total of 34% of amputees experienced a skin problem. Lesions resulting from friction, pressure, and occlusion are common. Allergic contact dermatitis is seen in a third of patients with stump dermatitis. There are no features that distinguish allergic from irritant (chemical or physical) dermatitis. CONCLUSION: Dermatologic problems are common in prosthetic limb users. Allergic contact dermatitis is a significant problem, and all patients with dermatitis on the residual limb should be patch tested. PMID- 10688726 TI - Surgical pearl: prompt treatment of subungual hematoma by decompression. PMID- 10688728 TI - Madelung's disease involving the tongue. AB - Madelung's disease or benign symmetric lipomatosis is rare. Symmetric lipomatosis of the tongue as an association of Madelung's disease is very rare; up to now only two cases of this association have been reported. We report the third case of Madelung's disease with involvement of the tongue in the form of macroglossia. PMID- 10688729 TI - Pemphigus foliaceus successfully treated with mycophenolate mofetil as a steroid sparing agent. AB - Pemphigus foliaceus is an autoimmune blistering disease of unknown origin with antibodies produced against desmoglein 1, an adhesive protein found in the desmosomal cell junction in the suprabasal layers of the epidermis. The disease is primarily treated with corticosteroids and corticosteroid-sparing immunosuppressive agents. We report a case of pemphigus foliaceus successfully treated with mycophenolate mofetil. It remains to be seen whether this agent has a significant effect on the course of the disease and remission induction. PMID- 10688730 TI - Linear verrucous hemangioma. AB - A 16-year-old male patient had multiple angiokeratotic lesions arranged in a linear pattern on his left arm. Histopathologic examination showed characteristic features of verrucous hemangioma. This entity should be distinguished from angiokeratoma or simple hemangioma. The linear arrangement of lesions as observed in this case may reflect genetic mosaicism. Deep surgery is the best treatment for verrucous hemangioma. PMID- 10688731 TI - Scleroderma-like reaction induced by uracil-tegafur (UFT), a second-generation anticancer agent. AB - UFT, a combination of uracil and tegafur, is a second-generation anticancer agent. UFT has been used in Japan, other Asian countries, South America, and Russia. Recently, UFT has been extensively studied for colorectal, pancreatic, and various types of cancer in North America and Europe, especially with leukovorin. We report a case of a scleroderma-like reaction induced by long-term administration of UFT. This is the first report of UFT-induced scleroderma-like reaction. PMID- 10688732 TI - A case of ichthyosis linearis circumflexa successfully treated with topical tacrolimus. AB - We report a case of ichthyosis linearis circumflexa (ILC) without the typical atopic manifestations and deformities of the hair shaft. The patient responded positively to treatment with topical tacrolimus, suggesting that abnormalities in the immunoregulatory mechanism may be involved in the pathogenesis of ILC. PMID- 10688733 TI - Squamous cell carcinoma, not basal cell carcinoma, is the most common cancer in humans. PMID- 10688734 TI - Sentinel lymph node biopsy as an adjunct to management of histologically difficult to diagnose melanocytic lesions: a proposal. AB - There is a significant subset of primary cutaneous melanocytic neoplasms that are difficult to diagnose with the use of routine light microscopy. The currently recommended approach in assessing such lesions is to make a histopathologic diagnosis that reflects some uncertainty and then to recommend complete surgical excision. While adequate in many cases, the excision that might be recommended for such a lesion if malignant would be mutilating in many others. To increase the sensitivity of diagnosis and to provide potentially useful prognostic information, we propose that sentinel lymphadenectomy be considered in patients with melanocytic neoplasms of uncertain behavior that are 1.0 mm or more in thickness. PMID- 10688735 TI - Liver biopsies and methotrexate: a time for reconsideration? PMID- 10688736 TI - Does Helicobacter pylori eradication treatment reduce the severity of rosacea? PMID- 10688738 TI - Helicobacter pylori: related to rosacea? PMID- 10688740 TI - Does Helicobacter pylori have a role in the pathogenesis of rosacea? PMID- 10688744 TI - Role of the development scientist in compound lead selection and optimization. AB - The R&D process for bringing drugs from discovery laboratories to the marketplace is undergoing rapid change, as enabled by new technologies and as demanded by the global pharmaceutical business environment. One consequence of the accelerated R&D paradigm is a blurring of the traditional discovery-development interface, which in turn impacts the traditional roles of discovery and development scientists. R&D organizations must find ways to screen out rapidly compounds that have relatively poor probability of successful registration. Quality of development candidates can be favorably influenced by early consideration of "developability" criteria along with receptor-based potency and specificity. Computational approaches and/or high-throughput experimental determinations will be used increasingly to profile compound characteristics which influence "developability." If such criteria are considered at the time of lead selection and optimization, the compound attrition rate during later development should be decreased from the historical norm. This article discusses the emerging role of development scientists during small-molecule lead selection and optimization. The changing role of development scientists also has implications for graduate curricula in the pharmaceutical sciences. PMID- 10688745 TI - Inter-study variability in population pharmacokinetic meta-analysis: when and how to estimate it? AB - Population pharmacokinetic analysis is being increasingly applied to individual data collected in different studies and pooled in a single database. However, individual pharmacokinetic parameters may change randomly from one study to another. In this article, we show by simulation that neglecting inter-study variability (ISV) does not introduce any bias for the fixed parameters or for the residual variability but may result in an overestimation of inter-individual (IIV) variability, depending on the magnitude of the ISV. Two random study-effect (RSE) estimation methods were investigated: (i) estimation, in a single step, of the three-nested random effects (inter-study, inter-individual and residual variability); (ii) estimation of residual variability and a mixture of ISV and IIV in the first step, then separation of ISV from IIV in the second. The one stage RSE model performed well for population parameter assessment, whereas, the two-stage model yielded good estimates of IIV only with a rich sampling design. Finally, irrespective of the method used, ISV estimates were valid only when a large number of studies was pooled. The analysis of one real data set illustrated the use of an ISV model. It showed that the fixed parameter estimates were not modified, whether an RSE model was used or not, probably because of the homogeneity of the experimental designs of the studies, and suggest no study effect in this example. PMID- 10688746 TI - Systemic availability and lymphatic transport of human growth hormone administered by subcutaneous injection. AB - Degradation of human growth hormone (hGH) at the injection site has previously been implicated as the basis for its reduced systemic availability following subcutaneous (SC) administration. The goal of these studies was to develop an animal model which would allow mass balance calculations to (i) quantify the loss at the injection site and (ii) determine the role of the lymphatics in the transport of subcutaneously-administered hGH. The animal model utilized a sheep and enabled simultaneous sampling of blood and collection of either peripheral lymph (via the efferent duct of the popliteal lymph node draining the injection site) or central lymph (via the thoracic lymph duct). In non-lymph cannulated sheep, the systemic availability of hGH following SC dosing was 58.4 +/- 9.1% (mean +/- SEM) relative to an intravenous (IV) control. The availability of hGH decreased to approximately 30-40% when either peripheral or central lymph was collected indicating that a proportion of the dose was transported via the lymph. The fraction of the administered dose collected in peripheral lymph was 61.7 +/- 8.5% (mean +/- SEM), whereas only 8.6 +/- 1.3% was collected in central lymph. These results suggested that loss of hGH within the lymphatics contributed significantly to its reduced systemic availability following SC administration. The total recovery (sum of the systemic availability and the cumulative amount recovered in lymph) of hGH was approximately 93% of the dose in the peripherally cannulated group indicating that loss at the injection site was minimal. PMID- 10688747 TI - Characterization of wet masses of pharmaceutical powders by triaxial compression test. AB - The mechanical and rheologic properties of wet masses of pharmaceutical powders determine their processibility and the quality of the product prepared by extrusion/spheronization. In this work, a triaxial compression test was attempted for the first time to characterize material properties of pharmaceutical wet masses of different hydrophilicity and particle sizes. The stress-strain curves and the pore pressure were determined at various cell pressures. The failure criteria of the wet masses were obtained from the stress path on the deviator stress plane. The cohesion (c) and the angle of internal friction (phi) were evaluated from the intercept and the slope of the failure loci. The stress-strain behavior strongly depended on the type of powders and cell pressure. The values of c and phi were similar for wet masses of EC FP, MCC PH101, and SMCC 50, but a very small phi and a very high c value for HPMC. The shear strength and rigidity of the wet masses were in the order of EC FP > SMCC 50 > MCC PH101 > HPMC, whereas the elastic recovery was in the opposite order. These material parameters could be used as references for selection of excipients and formulation for extrusion/spheronization. PMID- 10688748 TI - Isolation and identification of metabolites of porfiromycin formed in the presence of a rat liver preparation. AB - The isolation and identification of the major metabolites of porfiromycin formed in the presence of a rat liver preparation under aerobic conditions were performed with high-performance liquid chromatography and electrospray ionization mass spectrometry. Porfiromycin was extensively metabolized by the rat liver preparation in an aqueous 0.1 M potassium phosphate buffer (pH 7.4) containing an NADPH generating system at 37 degrees C. A total of eight metabolites was identified as mitosene analogs. Of these, three primary metabolites are 2 methylamino-7-aminomitosene, 1,2-cis and 1,2-trans-1-hydroxy-2-methylamino-7 aminomitosene, which are consistent with those previously observed in hypoxia using purified rat liver NADPH-cytochrome c reductase. Interestingly, 2 methylamino-7-aminomitosene is a reactive metabolite, which undergoes further activation at the C-10 position by the loss of carbamic acid and then links with the 7-amino group of the primary metabolites to yield two dimeric adducts. In addition, three phosphate adducts, 10-decarbamoyl-2-methylamino-7-aminomitosene 10-phosphate, 1,2-cis and 1,2-trans-2-methylamino-7-aminomitosene-1-phosphate, were also identified in the incubation system. The configurations of the diastereoisomeric metabolites were determined with (1)HNMR and phosphatase digestion. On the basis of the metabolite profile, we propose in vitro metabolic pathways for porfiromycin. The findings provide direct evidence for understanding the reactive nature and hepatic metabolism of the drug currently in phase III clinical trials. PMID- 10688749 TI - Optimization of storage stability of lyophilized actin using combinations of disaccharides and dextran. AB - The storage stability of a dry protein depends on the structure of the dried protein, as well as on the storage temperature relative to the glass transition temperature of the dried preparation. Disaccharides are known to preserve the native conformation of a dried protein; however, the resulting T(g) of the sample may be too low ensure adequate storage stability. On the other hand, formulations dried with high molecular weight carbohydrates, such as dextran, have higher glass transition temperatures, but fail to preserve native protein conformation. We tested the hypothesis that optimizing both protein structure and T(g) by freeze-drying actin with mixtures of disaccharides and dextran would result in increased storage stability compared to actin dried with either disaccharide or dextran alone. Protein structure in the dried solid was analyzed immediately after lyophilization and after storage at elevated temperatures with infrared spectroscopy, and after rehydration by infrared and circular dichroism spectroscopy. Structural results were related to the polymerization activity recovered after rehydration. Degradation was noted with storage for formulations containing either sucrose, trehalose, or dextran alone. Slight increases in T(g) observed in trehalose formulations compared to sucrose formulations did not result in appreciable increases in storage stability. Addition of dextran to sucrose or trehalose increased formulation T(g) without affecting the capacity of the sugar to inhibit protein unfolding during lyophilization and resulted in improved storage stability. Also, dextran provides an excellent amorphous bulking agent, which can be lyophilized rapidly with formation of strong, elegant cake structure. These results suggest that the strategy of using a mixture of disaccharide and polymeric carbohydrates can optimize protein storage stability. PMID- 10688750 TI - Studies on the structure of the complex of the boron neutron capture therapy drug, L-p-boronophenylalanine, with fructose and related carbohydrates: chemical and 13C NMR evidence for the beta-D-fructofuranose 2,3,6-(p phenylalanylorthoboronate) structure. AB - The complex of L-L-boronophenylalanine (L-p-BPA) with fructose has been used for the past 5 years in clinical trials of boron neutron capture therapy to treat both melanoma and glioblastoma multiforme. However, the structure of this complex in water buffered at physiologic pH has not been established. In the (1)H NMR spectra (D(2)O buffered at pD 7.4) of the complex of L-p-BPA with various carbohydrates, the upfield chemical shifts of the aromatic protons of L-p-BPA confirm that the boron atom is negatively charged and tetrahedral. In the (13)C NMR spectrum of the complex of L-p-BPA with U-(13)C labeled fructose, the chemical shifts and (1)J(CC) coupling constants are consistent with fructose adopting the beta-D-fructofuranose form. In addition, the (1)J(CC) coupling constants along with the binding constants measured for L-p-BPA with a series of monosaccharides and disaccharides seem to suggest that the beta-D-fructofuranose 2,3,6-(p-phenylalanylorthoboronate) structure strongly predominates, with free L p-BPA and fructose the only other species detected. PMID- 10688751 TI - Use of alveolar cell monolayers of varying electrical resistance to measure pulmonary peptide transport. AB - The apparent permeability coefficient (P(app)) of two fluorescently tagged model hydrophilic peptides, acXASNH(2) and acXAS(GAS)(7)NH(2), and (14)C-mannitol across monolayers of cultured rat alveolar epithelial cells of varying transepithelial electrical resistance (TER) has been examined. In line with their design features, the peptides were not degraded under the conditions of the test. Furthermore, no concentration dependence of transport of the tripeptide acXASNH(2) was observed over the concentration range studied, nor was any directional transport seen for either of the model peptides, indicating that under the conditions of the test they were not substrates for any transporters or efflux pumps. From the hydrophilic nature of the peptides (as assessed by their log P), and their inverse dependence of transport with molecular weight and TER, it was assumed that the peptides were transported across the cell monolayer passively via the paracellular route. The observed P(app) for the transport of (14)C-mannitol and the peptides across rat alveolar epithelial cell monolayers were found to be inversely (though not linearly) related to the measured TER and could be well-modeled assuming the presence of two populations of "pores" in the cell monolayer, namely, cylindrical pores of diameter 1.5 nm and large pores of diameter 20 nm. The relative populations of the two types of pores varied with the TER of the monolayer, with the number of large pores decreasing with an increase in TER (and the number of small pores taken as fixed). These results suggest that if the cell monolayer is well characterized with respect to the passage of a range of probe molecules across monolayers of varying electrical resistance, it should be possible to predict the P(app) of any hydrophilic peptide or drug crossing the membrane by the paracellular route at any desired TER using a monolayer of any electrical resistance, above a minimum value. PMID- 10688752 TI - Physico-chemical characterization of a novel tricyclic beta-lactam antibiotic. AB - GV118819X, a novel tricyclic beta-lactam antibiotic of GlaxoWellcome, is a racemic mixture of two diastereoisomers, A and B. Of the two diastereoisomers, only A is available as a pure compound. By analyzing mixtures of GV118819X and A, a partial phase diagram is constructed, which indicates the presence of a eutectic when the A fraction is approximately 39%. Moreover, the melting enthalpies of the eutectic mixture and of diastereoisomer B can be estimated. With the exception of the pure A form, all mixtures undergo important modifications in morphology and microstructure as a consequence of thermal treatments, which induce melting/amorphization of the eutectic, and crystallization of the A form. Analyses of the sieved fractions of GV118819X demonstrate that it consists of acicular crystals of different composition, with the larger crystals having a larger A fraction than the smaller ones. Grinding causes melting/amorphization of the eutectic and, following hours-long treatments, the formation of a substantial fraction of submicron particles with unusually low melting temperatures. PMID- 10688753 TI - Cyclodextrin-catalyzed deacetylation of spironolactone is pH and cyclodextrin dependent. AB - The complexation of spironolactone (SP) with cyclodextrins (CDs) and the effect of pH on the CD catalyzed deacetylation of SP was studied in the presence of beta cyclodextrin (beta-CD), hydroxypropyl-beta-cyclodextrin (HP-beta-CD), sulfobutylether-beta-cyclodextrin ([SBE](7m)-beta-CD), gamma-cyclodextrin (gamma CD), and sulfobutylether gamma-cyclodextrin (SBE-gamma-CD). The complexation of SP with beta-CD and the mechanism of deacetylation was confirmed using NMR. The complexation of SP with CDs was determined by means of the phase-solubility method at pH 2, in which chemical degradation was minimal. The phase-solubility diagrams were classified as A(L)-type and the apparent stability constants (K(1:1)) for 1 : 1 inclusion complex were calculated to be 9939 M(-1), 10,976 M( 1), 15,816 M(-1), 4792 M(-1) and 4118 M(-1) for beta-CD, HP-beta-CD, (SBE)(7m) beta-CD, gamma-CD, and SBE-gamma-CD, respectively. The effect of pH on the degradation rate of SP was studied in the presence and absence of 4.4 mM CD solutions at pH 4, 5, 6, 7, and 8 (25 degrees C). The stability studies showed that CD-catalyzed degradation of SP can be decreased by lowering the pH. The pH rate profiles of SP degradation with different CDs gave slopes of 1.0. Because no buffer catalysis was observed, the reaction appears to be specific-base catalyzed. The catalytic activity of CDs was as follows: SBE-gamma-CD < (SBE)(7m) beta-CD < HP-beta-CD approximately gamma-CD < beta-CD. NMR studies confirmed that SP forms an inclusion complex with beta-CD and complexation occurs by means of the secondary face. The NMR studies also showed that during the deacetylation of SP, the secondary hydroxyl groups of beta-CD at the 2- and 3-position were acetylated. The decrease of catalytic activity of CDs at low pH values and the CDs differing ability to catalyze the degradation of SP correlated qualitatively with the ionization state of the CD hydroxyl groups, which were lower in SBE-CDs. The site of binding differences and the number of hydroxyl groups present probably also contribute to the differences. PMID- 10688754 TI - Spherical crystal agglomeration of ibuprofen by the solvent-change technique in presence of methacrylic polymers. AB - The effects of Eudragit(R) nature on the formation and spherical agglomeration of ibuprofen microcrystals have been examined when solvent change (ethanol-water) technique is applied. Four methacrylic polymers (Eudragit(R) S100, L100, RS, and RL), with different solubility and solubilizing ability, were used. The extrapolated points of maximum temperature deviation rate in crystallization liquid that reflect the maximum crystallization rate and the corresponding water addition were determined, as well as crystal yielding and incorporation of drug and polymer in the agglomerates. The physicomechanical properties of the agglomerates, such as size, sphericity, surface roughness and porosity, as well as flow and packing or compression behavior during tableting, were evaluated for different drug/polymer ratios. It was found that crystal yield is greatly reduced in the presence of water-insoluble polymers and that formation of the microcrystals and incorporation of drug and polymer are affected by the polymer nature. Crystal formation changes are attributed to alterations in the metastable zone, whereas the changes in drug and polymer incorporation and crystal yield are caused by changes in the polymers' solubility and micellization. The size of agglomerates depends on the polymer nature and its interactions with the ibuprofen microcrystals formed. Sphericity, surface roughness, and intraparticle porosity of agglomerates increase, in general, with the presence of polymer owing to changes in habit and growth rate of the microcrystals and to their coating before binding into spherical agglomerates. The particle density or intraparticle porosity and size changes determine flow or packing behavior and densification of agglomerates at low compression. The incorporation and brittleness of the polymer determine the deformation under higher compression pressure, expressed as yield pressure, Py. PMID- 10688755 TI - Kinetic study of a 2-hydroxypropyl-beta-cyclodextrin-based formulation of all trans-retinoic acid in Sprague-Dawley rats after oral or intravenous administration. AB - all-trans-Retinoic acid (ATRA, vitamin A acid, or tretinoin) is a potent chemotherapeutic agent for the treatment of acute promyelocytic leukemia (APL). Its poor aqueous solubility not only affects its oral absorption but also prevents it from forming an aqueous parenteral formulation. Recently, we developed a water-soluble formulation of ATRA with 2-hydroxypropyl-beta cyclodextrin (HPbetaCD). In present study, this formulation was tested in Sprague Dawley rats. Kinetic study of ATRA was carried out after oral or intravenous administration. Though there were no statistical differences in any of the estimated pharmacokinetic parameters between ATRA sodium salt and HPbetaCD-based ATRA after intravenous administration, inclusion of ATRA into HPbetaCD was found to greatly improve the oral absorption of ATRA. PMID- 10688756 TI - Dissolution of ionizable water-insoluble drugs: the combined effect of pH and surfactant. AB - This study reports the results of the combined effect of pH and surfactant on the dissolution of piroxicam (PX), an ionizable water-insoluble drug in physiological pH. The intrinsic dissolution rate (J(total)) of PX was measured in the pH range from 4.0 to 7.8 with 0%, 0.5%, and 2.0% sodium lauryl sulfate (SLS) using the rotating disk apparatus. Solubility (c(total)) was also measured in the same pH and SLS concentration ranges. A simple additive model including an ionization (PX <--> H(+) + PX(-)) and two micellar solubilization equilibria (PX + micelle <--> [PX](micelle), PX(-) + micelle <--> [PX(-)](micelle)) were considered in the convective diffusion reaction model. J(total) and c(total) of PX increased with increasing pH and SLS concentration in an approximately additive manner. Nonlinear regression analysis showed that observed experimental data were well described with the proposed model (r(2) = 0.86, P < 0.001 for J(total) and r(2) = 0.98, P < 0.001 for c(total)). The pK(a) value of 5.63 +/- 0.02 estimated from c(total) agreed well with the reported value. The micellar solubilization equilibrium coefficient for the unionized drug was estimated to be 348 +/- 77 L/mol, while the value for the ionized drug was nearly equal to zero. The diffusion coefficients of the species PX, PX(-), and [PX](micelle) were estimated from the experimental results as (0. 93 +/- 0.35) x 10(-5), (1.4 +/- 0.30) x 10( 5), and (0.59 +/- 0.21) x 10(-5) cm(2)/s, respectively. The total flux enhancement is less than the total solubility enhancement due to the smaller diffusion coefficients of the micellar species. This model may be useful in predicting the dissolution of an ionizable water insoluble drug as a function of pH and surfactant and for establishing in vitro-in vivo correlations, IVIVC, for maintaining bioequivalence of drug products. PMID- 10688757 TI - Comparative effects of (SBE)7m-beta-CD and HP-beta-CD on the stability of two anti-neoplastic agents, melphalan and carmustine. AB - The purpose of this study was to evaluate and compare the potential use of two parenterally safe beta-cyclodextrins derivatives, (SBE)7m-beta-CD and HP-beta-CD, as solubilizers and stabilizers for melphalan and carmustine, two very unstable antineoplastic agents. Phase solubility and chemical stability of the compounds in the presence of the cyclodextrins were studied. UV, fluorescence, and several NMR techniques were used to probe the potential causes for the differences observed. The phase solubility method was found to provide only qualitative data on the binding of melphalan to the cyclodextrins since rapid degradation and the presence of products of degradation complicated the interpretation of the results. Qualitatively, however, the solubilizing potential was similar for the two cyclodextrins. The chemical stability studies indicate that both of the drugs had similar binding constants for both cyclodextrins; however, the intrinsic reactivities in the complexes were significantly lower with (SBE)7m-beta-CD than for HP-beta-CD. The main cause for this distinct difference appeared to correlate with differences in the site of binding and the polarity of the binding site. PMID- 10688758 TI - Editorial PMID- 10688759 TI - Halo Profiles and the Nonlinear Two- and Three-Point Correlation Functions of Cosmological Mass Density. AB - We investigate the nonlinear two- and three-point correlation functions of the cosmological density field in Fourier space and test the popular hierarchical clustering model that assumes a scale-independent three-point amplitude Q. In high-resolution N-body simulations of both the n=-2 scale-free model and the cold dark matter model, we find that Q at late times is not constant but increases with wavenumber far into the nonlinear regime. Self-similar scaling also does not hold as rigorously for the three-point function as for the two-point function in the n=-2 simulation; this is perhaps a manifestation of the finite simulation volume. We suggest that a better understanding of the behavior of the two- and three-point correlation functions in the nonlinear regime lies in the link to the density profiles of dark matter halos. We demonstrate and quantify how the slopes of the correlation functions are affected by the slope of the halo profiles using simple halo shapes and analytic clustering models. PMID- 10688760 TI - Does Deuterium Enable the Formation of Primordial Brown Dwarfs? AB - We investigate thermal and dynamical evolution of a primordial gas cloud with an updated deuterium chemistry. We consider a fragment of a postshock-cooled sheet that is expected to form by collapse of a massive cloud ( greater, similar108 M middle dot in circle) and by blast waves due to supernova explosions. At first we investigate molecule formation in a primordial shock. We show that almost all deuterium can be converted to HD within the age of the universe at the collapsed redshift in the case of a cloud that has a virial temperature of approximately 106 K and collapses at z>1. When the postshock sheet fragments owing to gravitational instability, the fractional H2 and HD abundances become approximately 10-2 and approximately 10-5, respectively, which are 103-104 times higher than the result of molecule formation in the expanding universe after recombination. To study the subsequent evolution of a fragment, we performed one dimensional simulations of a spherical/cylindrical cloud, of which initial conditions (e.g., fractional abundances of chemical composition, temperature) are derived from the result of the shock. It is found that, in case of a cylindrical collapse, the cooling by HD molecules keeps the temperature of the cloud less than 100 K and the cloud evolves almost isothermally. When the cloud becomes optically thick to the HD line emission ( approximately 1010 cm-3) and the gravitational fragmentation of the cylindrical cloud becomes effective, the Jeans mass becomes comparable to 0.1 M middle dot in circle. This series of processes enables the formation of primordial low-mass stars, and possibly brown dwarfs, in primordial gas clouds. PMID- 10688761 TI - Detection of an X-Ray Hot Region in the Virgo Cluster of Galaxies with ASCA. AB - Based on mapping observations with ASCA, an unusual hot region with a spatial extent of 1 deg2 was discovered between M87 and M49 at a center coordinate of R.A.=12h27m36s and decl.=9&j0;18' (J2000). The X-ray emission from the region has a 2-10 keV flux of 1x10-11 ergs s-1 cm-2 and a temperature of kT greater, similar4 keV, which is significantly higher than that in the surrounding medium of approximately 2 keV. The internal thermal energy in the hot region is estimated to be VnkT approximately 1060 ergs with a gas density of approximately 10-4 cm-3. A power-law spectrum with a photon index of 1.7-2.3 is also allowed by the data. The hot region suggests there is an energy input due to a shock that is probably caused by the motion of the gas associated with M49, infalling toward the M87 cluster with a velocity greater, similar1000 km s-1. PMID- 10688762 TI - Evidence for Stellar Streaming in the Cores of Elliptical Galaxies: A Kinematic Signature of Mergers? AB - We present evidence for non-Gaussian velocity fields within the cores of luminous elliptical galaxies. This evidence is based on high signal-to-noise ratio, medium resolution spectroscopy of the cores of early-type members of the Virgo and Coma Clusters obtained with the Wisconsin-Indiana-Yale-NOAO 3.5 m telescope. The Virgo data were acquired using an integral-field unit (DensePak), which allows the velocity field to be sampled over a variety of spatial scales. The Coma data were obtained through single 2&arcsec; diameter fibers. The cross-correlation profiles of luminous elliptical galaxies show considerable structure, often having several features with amplitudes as high as 10% that of the cross-correlation peak itself. This structure is most obvious within a radius of 1&farcs;5 (at Virgo), or GCP-2 > NAP-2. By the use of pertussis toxin (PTx), it was demonstrated that the actual events of G(alphai)-coupling to CXCR2 do not have a major role in the regulation of its internalization. Rather, CXCR2 internalization was shown to be negatively controlled by induction of signaling events, as indicated by the promotion of CXCR2 internalization following exposure to wortmannin, a potent inhibitor of phosphatidylinositol (PI) 3 kinases and PI4 kinases. Furthermore, our results suggest that rab11(+)-endosomes participate in the trafficking of CXCR2 through the endocytic pathway, to eventually allow its recycling back to the plasma membrane. To conclude, our findings shed light on the interrelationships between GCP-2 and other ELR(+)-CXC chemokines, and determine the mechanisms involved in the regulation of GCP-2-induced internalization and recycling of CXCR2. (Blood. 2000;95:1551-1559) PMID- 10688808 TI - Association between diabetic retinopathy and genetic variations in alpha2beta1 integrin, a platelet receptor for collagen. AB - Platelets might be involved in the pathogenesis of diabetic microangiopathy. Wide interindividual variations in the density of a platelet collagen receptor (alpha2beta1 integrin or glycoprotein Ia/IIa) are reportedly associated with polymorphism(s) in the gene encoding the alpha subunit of the receptor, including a Bgl II polymorphism in intron 7. The aim of the present study was to determine the relationship between the Bgl II polymorphism and the susceptibility to diabetic microangiopathy. A case-control study comparing 227 patients with type II diabetes mellitus (119 with versus 108 without diabetic retinopathy) as well as 169 nondiabetic subjects demonstrated that genotypes with Bgl II (+) allele had a significant increase in the risk for retinopathy. The odds ratio for Bgl II (+/+) to Bgl II (-/-) was 3.41 (95% CI, 1.49-7.78, P =.0036) when analysis was confined to those with a disease duration of diabetes of 10 years or more. The present study suggests that the presence of a Bg II (+) allele is a genetic risk factor for diabetic retinopathy. (Blood. 2000;95:1560-1564) PMID- 10688809 TI - Hemochromatosis genes and other factors contributing to the pathogenesis of porphyria cutanea tarda. AB - Inherited and acquired factors have been implicated in the pathogenesis of porphyria cutanea tarda (PCT), a disorder characterized by a photosensitive dermatosis and hepatic siderosis. This study, comprising 108 patients with PCT, was intended to define the role of hemochromatosis gene (HFE) mutations in the expression of PCT and to determine the contribution of acquired factors including alcohol, hepatitis C virus (HCV), and estrogen. The 2 known HFE mutations, cysteine 282 tyrosine (Cys282Tyr) and histidine 63 asparagine (His63Asp), were detected by polymerase chain reaction, and anti-HCV immunoglobulin G was detected serologically. Liver biopsies were graded for iron content, inflammation, and fibrosis. Estimates of alcohol and estrogen use were based on a questionnaire. Of the PCT patients tested, 19% were homozygous for the Cys282Tyr mutation; controls were equal to 0.5%. The compound heterozygous genotype was detected in 7% of the PCT patients; controls were less than 1%. The transferrin saturation, serum ferritin, and liver iron burden of all PCT patients were higher than those of nonporphyric controls. The highest values were found in PCT patients homozygous for the Cys282Tyr mutation. Of the patients studied, 59% were HCV positive (compared with 1.8% of the population), and 46% consumed more than 70 g of alcohol daily. Of the female patients, 63% were ingesting estrogens. Hepatic damage was most marked in patients with the Cys282Tyr/Cys282Tyr genotype who had HCV and drank heavily. Homozygosity for the Cys282Tyr mutation and HCV are the greatest risk factors for expression of PCT, and in most patients, more than 1 risk factor was identified. It was common for patients with HCV to consume alcohol. Patients with PCT should be screened for HFE mutations and for HCV. (Blood. 2000;95:1565-1571) PMID- 10688810 TI - Graft versus host disease prophylaxis with low-dose cyclosporine-A reduces the risk of relapse in children with acute leukemia given HLA-identical sibling bone marrow transplantation: results of a randomized trial. AB - Leukemia relapse is a major cause of treatment failure for patients with acute leukemia given allogeneic bone marrow transplantation (BMT). This study evaluated whether a reduction of the dosage of cyclosporine-A (Cs-A) used for graft versus host disease (GVHD) prophylaxis could reduce relapse rate (RR) in children with acute leukemia given BMT. Fifty-nine children who had transplantation from HLA identical siblings were randomized to receive Cs-A intravenously at a dosage of 1 mg/kg/d (Cs-A1) or of 3 mg/kg/d (Cs-A3) until patients were able to tolerate oral intake. Subsequently, both groups received Cs-A orally at a dosage of 6 mg/kg/d, with discontinuation 5 months after BMT. The probability of developing grade II IV acute GVHD was 57% for the Cs-A1 group versus 38% for the Cs-A3 group (P =.06); the probability of developing chronic GVHD was 30% for the Cs-A1 group and 26% for the Cs-A3 group (P = NS). Three patients died of grade IV acute GVHD: 2 were in the Cs-A1 and the third in the Cs-A3 group. The RR was 15% for the Cs-A1 group and 41% for the Cs-A3 group (P =.034); 1-year transplant-related mortality estimates were 17% and 7%, respectively (P = NS). With a median observation time of 44 months from BMT, the 5-year event-free survival for children belonging to Cs-A1 and Cs-A3 groups was 70% and 51%, respectively (P =.15). Our data demonstrate that the use of low Cs-A doses is associated with a statistically significant reduction of leukemia relapse, probably due to an increased graft versus leukemia effect. (Blood. 2000;95:1572-1579) PMID- 10688811 TI - Patients with myelodysplastic syndromes benefit from palliative therapy with amifostine, pentoxifylline, and ciprofloxacin with or without dexamethasone. AB - Thirty-five patients with myelodysplastic syndrome (MDS) were registered on protocol MDS 96-02 and were receiving continuous therapy with pentoxifylline 800 mg 3 times a day and ciprofloxacin 500 mg twice a day by mouth; dexamethasone was added to the regimen for the partial responders and the nonresponders after 12 weeks at a dose of 4 mg by mouth every morning for 4 weeks. Amifostine was administered intravenously 3 times a week at 3 dose levels (200 mg/M(2), 300 mg/M(2), and 400 mg/M(2)) to cohorts of 10 patients each. Therapy has been continued for 1 year in responders. Twenty-nine have completed at least 12 weeks of therapy and are available for response evaluation. Of the 21 men and 8 women (median age, 67 years), 20 had refractory anemia (RA), 3 had RA with ringed sideroblasts (RARS), 5 had RA with excess blasts (RAEB), and 1 had chronic myelomonocytic leukemia (CMMoL). Five had secondary MDS. No differences were noted in response rates among the 3 dose levels. Seven patients did not respond at all, and 22 showed an improvement in cytopenias (76%). Three had a triple lineage response, 10 had a double lineage response, and 9 had a single lineage response (8 of 9 in absolute neutrophil count [ANC] and 1 had more than a 50% reduction in packed red blood cell transfusions). Fifteen patients responded only after the addition of dexamethasone, whereas 7 responded before. When examined by lineage, 19 of 22 showed improved ANC, 11 of 22 demonstrated more than 50% reduction in blood transfusions, improved Hb levels, or both, and 7 of 22 showed improvement in platelet counts. Interestingly, the responses were frequently slow to appear, and continued improvement in counts was seen up to 12 months of therapy and beyond. This study supports the feasibility of treating patients with MDS with the unique approach of cytoprotection and anticytokine therapies as well as the principle that prolonged commitment to treatment is desirable when noncytotoxic agents are administered. (Blood. 2000;95:1580-1587) PMID- 10688812 TI - Predictors of therapy-related leukemia and myelodysplasia following autologous transplantation for lymphoma: an assessment of risk factors. AB - We analyzed data on 612 patients who had undergone high-dose chemoradiotherapy (HDT) with autologous stem cell rescue for Hodgkin's disease (HD) and non Hodgkin's lymphoma (NHL) at the City of Hope National Medical Center, to evaluate the incidence of therapy-related myelodysplasia (t-MDS) or therapy-related acute myeloid leukemia (t-AML) and associated risk factors. A retrospective cohort and a nested case-control study design were used to evaluate the role of pretransplant therapeutic exposures and transplant conditioning regimens. Twenty two patients developed morphologic evidence of t-MDS/t-AML. The estimated cumulative probability of developing morphologic t-MDS/t-AML was 8.6% +/- 2.1% at 6 years. Multivariate analysis of the entire cohort revealed stem cell priming with VP-16 (RR = 7.7, P = 0.002) to be independently associated with an increased risk of t-MDS/t-AML. The influence of pretransplant therapy on subsequent t-MDS/t AML risk was determined by a case-control study. Multivariate analysis revealed an association between pretransplant radiation and the risk of t-MDS/t-AML, but failed to reveal any association with pretransplant chemotherapy or conditioning regimens. However, patients who had been primed with VP-16 for stem cell mobilization were at a 12. 3-fold increased risk of developing t-AML with 11q23/21q22 abnormalities (P = 0.006). Patients undergoing HDT with stem cell rescue are at an increased risk of t-MDS/t-AML, especially those receiving priming with VP-16 for peripheral stem cell collection. (Blood. 2000;95:1588 1593) PMID- 10688813 TI - Sustained expression of human factor VIII in mice using a parvovirus-based vector. AB - Persistent therapeutic levels of human factor VIII (hFVIII) would signify a major advance in the treatment of hemophilia A. Here we report sustained expression of hFVIII in immunocompetent mice using recombinant adeno-associated virus (rAAV) vectors. AAV can stably transduce liver cells, the target tissue for efficient hFVIII production. Because of rAAV packaging constraints, we tested 2 constructs using small regulatory elements designed for liver-specific transgene expression linked to B-domain-deleted hFVIII (BDD-hFVIII) cDNA. More than 10(12)/mL rAAV/BDD hFVIII virion particles were generated using a transfection scheme that eliminates adenovirus. Coatest and APTT assays confirmed the production of functional BDD-hFVIII protein after transduction of 293 and HepG2 cells. In vivo experiments were performed in C57BL/6 and NOD/scid mice receiving 10(10-11) rAAV/hFVIII particles via portal vein injection. All C57BL/6 mice tested developed anti-hFVIII antibody. In contrast, NOD/scid mice expressed hFVIII reaching 27% of normal human plasma levels. As expected, we could not detect hFVIII antigen from plasma samples isolated from control animals receiving equivalent doses of rAAV expressing enhanced green fluorescent protein (EGFP). Transgene mRNA expression was detected primarily in the liver and histologic analysis of the liver revealed no pathologic abnormalities. These results demonstrate a promising approach for treatment of hemophilia A. (Blood. 2000;95:1594-1599) PMID- 10688814 TI - Interleukin 5 regulates the isoform expression of its own receptor alpha-subunit. AB - The receptor for interleukin 5 (IL-5) consists of a cytokine-specific alpha chain (IL-5Ralpha) and a signaling beta chain, which is shared with interleukin 3 (IL 3) and granulocyte-macrophage colony-stimulating factor (GM-CSF). These 3 cytokines can act in eosinophil development and activation in vitro, but gene deletion or antibody blocking of IL-5 largely ablates eosinophilic responses in models of allergic disease or helminth infection. We investigated factors acting in differential IL-5Ralpha gene splicing to generate either the membrane-anchored isoform (TM-IL-5Ralpha) which associates with the common beta chain to allow IL-5 responsiveness, or a secreted, antagonist variant (SOL-IL-5Ralpha). In a murine myeloid cell line (FDC-P1), transfected with minigenes allowing expression of either IL-5Ralpha variant, IL-5 itself, but not IL-3 or GM-CSF, stimulated a reversible switch toward expression of TM-IL-5Ralpha. A switch from predominantly soluble isoform to TM-IL-5Ralpha messenger RNA (mRNA) expression was also seen during IL-5-driven eosinophil development from human umbilical cord blood-derived CD34(+) cells; this was accompanied by surface expression of IL-5Ralpha and acquisition of functional responses to IL-5. IL-3 and GM-CSF also supported eosinophil development and up-regulation of TM-IL-5Ralpha mRNA in this system, but this was preceded by expression of IL-5 mRNA and was inhibited by monoclonal antibody to IL-5. These data suggest IL-5-specific signaling, not shared by IL-3 and GM-CSF, leading to a switch toward up-regulation of functional IL-5Ralpha and, furthermore, that IL-3 and GM-CSF-driven eosinophil development is dependent on IL-5, providing an explanation for the selective requirement of IL-5 for expansion of the eosinophil lineage. (Blood. 2000;95:1600-1607) PMID- 10688815 TI - Spatial associations of centromeres in the nuclei of hematopoietic cells: evidence for cell-type-specific organizational patterns. AB - It is believed that the 3-dimensional organization of centromeric heterochromatin in interphase may be of functional relevance as an epigenetic mechanism for the regulation of gene expression. Accordingly, a likely possibility is that the centromeres that spatially associate into the heterochromatic structures (chromocenters) observed in the G1 phase of the cell cycle will differ in different cells. We sought to address this issue using, as a model, the chromocenters observed in quiescent normal human hematopoietic cells and primary fibroblasts. To do this, we analyzed the spatial relationships among different human centromeres in 3-D preserved cells using nonisotopic in situ hybridization and confocal microscopy. We showed quantitatively that chromocenters in all cell types do indeed represent nonrandom spatial associations of certain centromeres. Furthermore, the observed patterns of centromere association indicate that the chromocenters in these cell types are made of different combinations of specific centromeres, that hematopoietic cells are strikingly different from fibroblasts as to the composition of their chromocenters and that centromeres in peripheral blood cells appear to aggregate into distinct "myeloid" (present in monocytes and granulocytes) and "lymphoid" (present in lymphocytes) spatial patterns. These findings support the idea that the chromocenters formed in the nucleus of quiescent hematopoietic cells might represent heterochromatic nuclear compartments involved in the regulation of cell-type-specific gene expression, further suggesting that the spatial arrangement of centromeric heterochromatin in interphase is ontogenically determined during hematopoietic differentiation. (Blood. 2000;95:1608-1615) PMID- 10688816 TI - A soluble form of human Delta-like-1 inhibits differentiation of hematopoietic progenitor cells. AB - Two Notch ligand families, Delta and Serrate/Jagged, have been identified in vertebrates. Members of the Jagged family have been shown to affect in vitro hematopoiesis. To determine whether members of the Delta family might play a similar role in hematopoiesis, we examined the expression of mouse Delta-like-1 (mDll1). mDll1 protein was detected in whole marrow and in a marrow stromal cell line MS-5. At the RNA level, both mDll1 and Notch1 were seen in marrow precursor, differentiated hematopoietic, marrow stromal, and MS-5 cells. We isolated a cDNA encoding the human homologue of mDll1, designated human Delta-like-1 (hDll1). A soluble form of hDll1, hDll1(NDSL), containing the DSL domain and the N-terminal sequences, was expressed and purified from bacteria as a glutathione S transferase (GST) fusion protein. We observed that hDll1(NDSL) delayed the acquisition of differentiation markers by murine hematopoietic progenitor cells (Lin-) cultured in vitro with cytokines. In addition, it promoted greater expansion (more than 3 times) of the primitive hematopoietic precursor cell population, measured in high-proliferative potential colony assay and day 12 colony-forming unit spleen (CFU-S) assay, than GST controls. We also observed that the percentage of apoptotic cells decreased and that the number of cells in the S-phase of the cell cycle increased in the cultures of Lin(-) cells with hDll1(NDSL). The effects of hDll1(NDSL) were blocked by antibody against the mouse counterpart of hDll1(NDSL), mDll1(NDSL). These observations demonstrate that hDll1 plays a role in mediating cell fate decisions during hematopoiesis. (Blood. 2000;95:1616-1625) PMID- 10688817 TI - Regulation of Jak2 tyrosine kinase by protein kinase C during macrophage differentiation of IL-3-dependent myeloid progenitor cells. AB - Differentiation of macrophages from myeloid progenitor cells depends on a discrete balance between cell growth, survival, and differentiation signals. Interleukin-3 (IL-3) supports the growth and survival of myeloid progenitor cells through the activation of Jak2 tyrosine kinase, and macrophage differentiation has been shown to be regulated by protein kinase C (PKC). During terminal differentiation of macrophages, the cells lose their mitogenic response to IL-3 and undergo growth arrest, but the underlying signaling mechanisms have remained elusive. Here we show that in IL-3-dependent 32D myeloid progenitor cells, the differentiation-inducing PKC isoforms PKC-alpha and PKC-delta specifically caused rapid inhibition of IL-3-induced tyrosine phosphorylation. The target for this inhibition was Jak2, and the activation of PKC by 12-O-tetradecanoyl-phorbol-13 acetate treatment also abrogated IL-3-induced tyrosine phosphorylation of Jak2 in Ba/F3 cells. The mechanism of this regulation was investigated in 32D and COS7 cells, and the inhibition of Jak2 required catalytic activity of PKC-delta and involved the phosphorylation of Jak2 on serine and threonine residues by the associated PKC-delta. Furthermore, PKC-delta inhibited the in vitro catalytic activity of Jak2, indicating that Jak2 was a direct target for PKC-delta. In 32D cells, the inhibition of Jak2 either by PKC-delta, tyrosine kinase inhibitor AG490, or IL-3 deprivation caused a similar growth arrest. Reversal of PKC-delta mediated inhibition by the overexpression of Jak2 promoted apoptosis in differentiating 32D cells. These results demonstrate a PKC-mediated negative regulatory mechanism of cytokine signaling and Jak2, and they suggest that it serves to integrate growth-promoting and differentiation signals during macrophage differentiation. (Blood. 2000;95:1626-1632) PMID- 10688818 TI - Existence of a differentiation blockage at the stage of a megakaryocyte precursor in the thrombocytopenia and absent radii (TAR) syndrome. AB - The thrombocytopenia and absent radii (TAR) syndrome is a rare disease associating bilateral radial agenesis and congenital thrombocytopenia. Here, we investigated in vitro megakaryocyte (MK) differentiation and expression of c-mpl in 6 patients. Using blood or marrow CD34(+) cells, the colony-forming unit (CFU) MK number was markedly reduced. CD34(+) cells were also cultured in liquid medium in the presence of a combination of 3 cytokines (stem cell factor, interleukin-3, and interleukin-6) or megakaryocyte growth and development factor (PEG-rHuMGDF) with or without SCF. In the presence of PEG-rHuMGDF, the majority of mature megakaryocytes (CD41 high, CD42 high) underwent apoptosis. This phenomenon was also observed in cultures stimulated by three cytokines. However, this last combination of cytokines allowed a more complete terminal MK differentiation. Surprisingly, a homogeneous population of CD34(-)CD41(+)CD42(-) cells accumulated during the cultures. This population was unable to differentiate along the myeloid pathways. This result suggests that a fraction of MK cells is unable to differentiate in the TAR syndrome. We subsequently investigated whether this could be related to an abnormality in c-mpl. No mutation or rearrangement in the c-mpl gene was found by Southern blots or by sequencing of the c-mpl coding region and its promoter in any of the patients. Using Western blot analysis, a decreased level of Mpl was found in patient platelets. A decreased level of c-mpl messenger RNA in TAR platelets was also detected with a lower c-mpl-P to c-mpl-K ratio in comparison to adult platelets. Altogether, these results demonstrate that the thrombocytopenia of the TAR syndrome is associated with a dysmegakaryocytopoiesis characterized by cells blocked at an early stage of differentiation. (Blood. 2000;95:1633-1641) PMID- 10688819 TI - Leishmania donovani infection of bone marrow stromal macrophages selectively enhances myelopoiesis, by a mechanism involving GM-CSF and TNF-alpha. AB - Alterations in hematopoiesis are common in experimental infectious disease. However, few studies have addressed the mechanisms underlying changes in hematopoietic function or assessed the direct impact of infectious agents on the cells that regulate these processes. In experimental visceral leishmaniasis, caused by infection with the protozoan parasite Leishmania donovani, parasites persist in the spleen and bone marrow, and their expansion in these sites is associated with increases in local hematopoietic activity. The results of this study show that L donovani targets bone marrow stromal macrophages in vivo and can infect and multiply in stromal cell lines of macrophage, but not other lineages in vitro. Infection of stromal macrophages increases their capacity to support myelopoiesis in vitro, an effect mediated mainly through the induction of granulocyte macrophage-colony stimulating factor and tumor necrosis factor-alpha. These data are the first to directly demonstrate that intracellular parasitism of a stromal cell population may modify its capacity to regulate hematopoiesis during infectious disease. (Blood. 2000;95:1642-1651) PMID- 10688820 TI - The GATA-E box-GATA motif in the EKLF promoter is required for in vivo expression. AB - The erythroid Kruppel-like factor (EKLF) is a key regulatory protein in globin gene expression. This zinc finger transcription factor is required for expression of the adult beta globin gene, and it has been suggested that it plays an important role in the developmental switch from fetal gamma to adult beta globin gene expression. We have previously described a sequence element in the distal promoter region of the mouse EKLF gene that is critical for the expression of this transcription factor. The element consists of an E box motif flanked by 2 GATA-1 binding sites. Here we demonstrate that mutation of the E box or the GATA 1 consensus sequences eliminates expression from the EKLF promoter in transgenic mice. These results confirm the importance of this activator element for in vivo expression of the EKLF gene. (Blood. 2000;95:1652-1655) PMID- 10688821 TI - Activation of Akt kinase by granulocyte colony-stimulating factor (G-CSF): evidence for the role of a tyrosine kinase activity distinct from the Janus kinases. AB - Activation of the serine/threonine kinase Akt has been shown to be a critical component for growth factor and cytokine stimulation of cell survival. Although some of the immediate upstream activators of Akt have been defined, the roles of tyrosine kinases in the activation of Akt are not well delineated. Granulocyte colony-stimulating factor (G-CSF) regulates the proliferation, differentiation, and survival of neutrophilic granulocytes. G-CSF exerts its actions by stimulating several signaling cascades after binding its cell surface receptor. Both Jak (Janus) and Src families of tyrosine kinases are stimulated by incubation of cells with G-CSF. In this report, we show that G-CSF stimulation of cells leads to activation of Akt. The membrane-proximal 55 amino acids of the G CSF receptor cytoplasmic domain are sufficient for mediating Akt activation. However, activation of Akt appears to be downregulated by the receptor's carboxy terminal region of 98 amino acids, a region that has been shown to be truncated in some patients with acute myeloid leukemia associated with severe congenital neutropenia. Furthermore, we demonstrate that G-CSF-induced activation of Akt requires the activities of Src family kinases but can be clearly dissociated from G-CSF-stimulated activation of Stats (signal transducers and activators of transcription) by the Jak kinases. Thus, cytokine activation of the Jak/Stat and other signaling cascades can be functionally separated. (Blood. 2000;95:1656 1662) PMID- 10688822 TI - Rapid tyrosine phosphorylation and activation of Bruton's tyrosine/Tec kinases in platelets induced by collagen binding or CD32 cross-linking. AB - Stimulation of the platelet nonintegrin collagen receptor, glycoprotein VI, evokes a signaling response similar to that induced by antigen receptor activation in B and T lymphocytes. A key transducer of the lymphocyte signaling pathways is the Bruton's tyrosine kinase (Btk)/Tec kinase family, which connects receptors to the elevation of intracellular-free calcium levels. An important signaling function for Btk in collagen-induced platelet activation in vitro was recently demonstrated by other researchers using Btk-deficient platelets from patients with X-linked agammaglobulinemia (XLA). Since Btk-deficiency does not induce an overt platelet-based bleeding disorder in vivo, collagen receptor responses may include other Btk/Tec kinase family members in normal platelets. Both Btk and Tec had increased tyrosine following stimulation of collagen receptors or CD32 cross-linking. Data from kinetic analyses and inhibitor studies and the use of phosphopeptide-specific antibodies recognizing 2 Btk regulatory phosphorylated tyrosine residues suggest a mechanism for coordinate recruitment of Btk and Tec through the immunoreceptor tyrosine-based activation motif, Src family kinases, and phosphatidylinositol 3-kinase. In XLA platelets, collagen treatment increased tyrosine phosphorylation of Tec and several other signaling proteins, including Lyn, Fyb, Slp-76, and the Wiskott-Aldrich syndrome protein. This indicates that important elements of the collagen signaling pathway proximal and distal to Btk and Tec are preserved despite the lack of functional Btk. The results are consistent with the conclusion that activation of Tec may sustain XLA platelet function in vivo, while some in vitro assays of nonintegrin collagen receptor signaling through the Btk/Tec kinase family reflect the additive dosage of the transducers. (Blood. 2000;95:1663-1670) PMID- 10688823 TI - Vasculogenesis in the day 6.5 to 9.5 mouse embryo. AB - The process of vasculogenesis was characterized in the 6.5- to 9.5-day mouse embryo and in allantoic culture by analysis of spatial and temporal expression patterns of the endothelial or hematopoietic lineage-associated proteins, TAL1, Flk1, platelet/endothelial cell adhesion molecule (PECAM), CD34, VE-cadherin, and Tie2. The study establishes that: (1) TAL1 and Flk1 are coexpressed in isolated mesodermal cells that give rise to endothelial cells and thus can be defined as angioblasts; (2) hematopoietic cells of blood islands express TAL1, but not Flk1; (3) vasculogenesis in the embryo proper is initiated by mesoderm fated to give rise to the endocardium; (4) the maturation/morphogenesis of blood vessels can be defined in terms of a sequential pattern of expression in which TAL1 and Flk1 are expressed first followed by PECAM, CD34, VE-cadherin, and later Tie2; and (5) TAL1 expression is down-regulated in endothelial cells of mature vessels. (Blood. 2000;95:1671-1679) PMID- 10688824 TI - The endothelial cell protein C receptor aids in host defense against Escherichia coli sepsis. AB - The influence of the endothelial protein C receptor (EPCR) on the host response to Escherichia coli was studied. Animals were treated with 4 separate protocols for survival studies and analysis of physiologic and biochemical parameters: (1) monoclonal antibody (mAb) that blocks protein C/activated protein C binding to EPCR plus sublethal numbers of E coli (SLEC) (n = 4); (2) mAb to EPCR that does not block binding plus SLEC (n = 3); (3) SLEC alone (n = 4); and (4) blocking mAB alone (n = 1). Those animals receiving blocking mAb to EPCR plus sublethal E coli died 7 to 54 hours after challenge, whereas all animals treated with the other protocols were permanent survivors. Histopathologic studies of tissues from animals receiving blocking mAb plus SLEC removed at postmortem were compared with those animals receiving SLEC alone killed at T+24 hours. The animals receiving the blocking mAb exhibited consumption of fibrinogen, microvascular thrombosis with hemorrhage of both the adrenal and renal cortex, and an intense influx of neutrophils into the adrenal, renal, and hepatic microvasculature, whereas the tissues from animals receiving only sublethal E coli exhibited none of these abnormal histopathologic changes. Compared with the control animals, the animals receiving the blocking mAb exhibited significantly elevated serum glutamic pyruvic transaminase, anion gap, thrombin-antithrombin complex, IL-6, IL-8, and soluble thrombomodulin. The levels of circulating activated protein C varied too widely to allow a clear determination of whether the extent of protein C activation was altered in vivo by blocking protein C binding to EPCR. We conclude that protein C/activated protein C binding to EPCR contributes to the negative regulation of the coagulopathic and inflammatory response to E coli and that EPCR provides an additional critical step in the host defense against E coli. (Blood. 2000;95:1680-1686) PMID- 10688825 TI - Endotoxin and thrombin elevate rodent endothelial cell protein C receptor mRNA levels and increase receptor shedding in vivo. AB - The endothelial cell protein C receptor (EPCR) facilitates protein C activation by the thrombin-thrombomodulin complex. Protein C activation has been shown to be critical to the host defense against septic shock. In cell culture, tumor necrosis factor-alpha (TNF-alpha) down-regulates EPCR expression, raising the possibility that EPCR might be down-regulated in septic shock. We examined EPCR mRNA and soluble EPCR levels in mice and rats challenged with lethal dose 95 levels of endotoxin. Toxic doses of TNF-alpha failed to alter EPCR mRNA levels in mice. Rather than EPCR mRNA levels falling in response to endotoxin, as predicted from cell-culture experiments, they rose approximately 3-fold 6 hours after exposure to endotoxin before returning toward baseline levels at 24 hours after exposure. Soluble EPCR levels rose approximately 4-fold. Infusion of hirudin, a specific thrombin inhibitor, before endotoxin exposure almost completely blocked the increase in EPCR mRNA and soluble EPCR. Consistent with the idea that the responses were mediated by thrombin, thrombin infusion (5 U/kg of body weight for 3 hours) resulted in an approximately 2-fold increase in EPCR mRNA and soluble EPCR. Incubation of rat endothelial cells with thrombin or murine protease activated receptor 1 agonist peptide resulted in a 2-fold increase in EPCR mRNA. These results indicate that thrombin plays a major role in up-regulating EPCR mRNA and shedding in vivo. (Blood. 2000;95:1687-1693) PMID- 10688826 TI - Surface expression and functional characterization of alpha-granule factor V in human platelets: effects of ionophore A23187, thrombin, collagen, and convulxin. AB - Factor V (FV) present in platelet alpha-granules has a significant but incompletely understood role in hemostasis. This report demonstrates that a fraction of platelets express very high levels of surface-bound, alpha-granule FV on simultaneous activation with 2 agonists, thrombin and convulxin, an activator of the collagen receptor glycoprotein VI. This subpopulation of activated platelets represents 30.7% +/- 4.7% of the total population and is referred to as convulxin and thrombin-induced-FV (COAT-FV) platelets. COAT-FV platelets are also observed on activation with thrombin plus collagen types I, V, or VI, but not with type III. No single agonist examined was able to produce COAT-FV platelets, although ionophore A23187 in conjunction with either thrombin or convulxin did generate this population. COAT-FV platelets bound annexin-V, indicating exposure of aminophospholipids and were enriched in young platelets as identified by the binding of thiazole orange. The functional significance of COAT-FV platelets was investigated by demonstrating that factor Xa preferentially bound to COAT-FV platelets, that COAT-FV platelets had more FV activity than either thrombin or A23187-activated platelets, and that COAT-FV platelets were capable of generating more prothrombinase activity than any other physiologic agonist examined. Microparticle production by dual stimulation with thrombin and convulxin was less than that observed with A23187, indicating that microparticles were not responsible for all the activities observed. These data demonstrate a new procoagulant component produced from dual stimulation of platelets with thrombin and collagen. COAT-FV platelets may explain the unique role of alpha-granule FV and the hemostatic effectiveness of young platelets. (Blood. 2000;95:1694-1702) PMID- 10688827 TI - Involvement of low-density lipoprotein receptor-related protein (LRP) in the clearance of factor VIII in von Willebrand factor-deficient mice. AB - Factor VIII is tightly noncovalently linked to von Willebrand factor (vWF) in plasma with a stoichiometry of 1:50, and vWF deficiency results in secondary factor VIII deficiency, with accelerated clearance of factor VIII from the circulation. We used a murine model of severe von Willebrand disease (vWF knockout mice) to study the effect of a recombinant vWF/pro-vWF preparation (rpvWF) on factor VIII survival and to investigate whether low-density lipoprotein receptor-related protein (LRP) might be involved in the in vivo clearance of factor VIII in the absence of vWF. vWF-deficient mice received 70 U/kg rpvWF in the first series of experiments, and in a second series, 80 mg/kg receptor-associated protein (RAP) as a recombinant fusion protein to block the action of LRP. Factor VIII levels were measured at time 0, or 1 or 3 hours after administration of rpvWF or RAP. RAP induced a sustained rise in factor VIII levels comparable to that induced by rpvWF. In a third series, the preadministration of RAP resulted in a slower disappearance of factor VIII antigen (measured by an enzyme-linked immunosorbent assay specific for human factor VIII) after infusion of recombinant factor VIII. These findings suggest that the accelerated clearance of factor VIII seen in the absence of vWF may be a result of the involvement of LRP in factor VIII metabolism. (Blood. 2000;95:1703 1708) PMID- 10688828 TI - Hypofibrinogenemia in an individual with 2 coding (gamma82 A-->G and Bbeta235 P- >L) and 2 noncoding mutations. AB - We investigated the molecular basis of hypofibrinogenemia in a man with a normal thrombin clotting time. Protein analysis indicated equal plasma expression of 2 different Bbeta alleles, and DNA sequencing confirmed heterozygosity for a new Bbeta235 P-->L mutation. Protein analysis also revealed a novel gamma(D) chain, present at a ratio of 1:2 relative to the gamma(A) chain. Mass spectrometry indicated a 14 d decrease in the gamma(D)-chain mass, and DNA sequencing showed this was caused by a novel gamma82 A-->G substitution. DNA sequencing established heterozygosity for 2 further mutations: T-->C in intron 4 of the Aalpha gene and A-->C in the 3' noncoding region of the Bbeta gene. Studies on the man's daughter, together with plasma expression levels, discounted both the Aalpha and Bbeta mutations as the cause of the low fibrinogen, suggesting that the gamma82 mutation caused the hypofibrinogenemia. This was supported by analysis of 31 normal controls in whom the Bbeta mutations were found at polymorphic levels, with an allelic frequency of 5% for the Bbeta235 mutation and 42% for the Bbeta 3' untranslated mutation. The gamma82 mutation was, however, unique to the propositus. Residue gamma82 is located in the triple helix that separates the E and D domains, and aberrant packing of the helices may explain the decreased fibrinogen concentration. (Blood. 2000;95:1709-1713) PMID- 10688829 TI - Regulation of factor VIIIa by human activated protein C and protein S: inactivation of cofactor in the intrinsic factor Xase. AB - Factor VIIIa is a trimer of A1, A2, and A3-C1-C2 subunits. Inactivation of the cofactor by human activated protein C (APC) results from preferential cleavage at Arg336 within the A1 subunit, followed by cleavage at Arg562 bisecting the A2 subunit. In the presence of human protein S, the rate of APC-dependent factor VIIIa inactivation increased several-fold and correlated with an increased rate of cleavage at Arg562. (Active site-modified) factor IXa, blocked cleavage at the A2 site. However, APC-catalyzed inactivation of factor VIIIa proceeded at a similar rate independent of factor IXa, consistent with the location of the preferential cleavage site within the A1 subunit. Addition of protein S failed to increase the rate of cleavage at the A2 site when factor IXa was present. In the presence of factor X, cofactor inactivation was inhibited, due to a reduced rate of cleavage at Arg336. However, inclusion of protein S restored near original rates of factor VIIIa inactivation and cleavage at the A1 site, thus overcoming the factor X-dependent protective effect. These results suggest that in the human system, protein S stimulates APC-catalyzed factor VIIIa inactivation by facilitating cleavage of A2 subunit (an effect retarded in the presence of factor IXa), as well as abrogating protective interactions of the cofactor with factor X. (Blood. 2000;95:1714-1720) PMID- 10688830 TI - A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin: its application to the measurement of granulocyte-derived elastase digests in plasma. AB - When granulocytes are stimulated under certain clinical conditions, elastase is released therefrom and digests fibrin(ogen) independently of the plasmin system, which may also be mobilized simultaneously. Thus, discrimination of these 2 systems becomes urgent for the diagnosis and treatment of the underlying diseases. Using as immunogen a 97-kd granulocyte-elastase digest of human fibrinogen, we raised an antibody IF-123 that specifically recognizes elastase digests of human fibrin(ogen). The 97-kd elastase fragment resembles plasmic fragment D(1), and the epitope of this antibody is located on the Aalpha (196 204) residue segment. This segment appears to be masked in fibrin(ogen) but exposed when the Aalpha Leu 204-Ile 205 peptide bond is cleaved by elastase. Cathepsin G concomitantly released from granulocytes failed to expose the epitope. By an enzyme immunoassay using IF-123 as the capture antibody, the elastase digests of fibrin(ogen) can be measured in plasma samples without interference by abundantly coexisting fibrinogen. Indeed, we found that the elastase digests were mostly elevated in patients with inflammation or malignant tumors, but remained in a normal range in patients with a benign gastrointestinal tract disease such as duodenal ulcer and polyps in the gallbladder or the colon. Like the plasmic D-dimer, the elastase digests predominantly consisted of the DD/E complex and DD/E-containing high-molecular weight derivatives apparently corresponding to the phase-3 plasmic digests of cross-linked fibrin. (Blood. 2000;95:1721-1728) PMID- 10688831 TI - Lepirudin blunts endotoxin-induced coagulation activation. AB - During sepsis, lipopolysaccharide (LPS) triggers the development of disseminated intravascular coagulation (DIC) via the tissue factor-dependent pathway of coagulation resulting in massive thrombin generation and fibrin polymerization. Recently, animal studies demonstrated that hirudin reduced fibrin deposition in liver and kidney and decreased mortality in LPS-induced DIC. Accordingly, the effects of recombinant hirudin (lepirudin) was compared with those caused by placebo on LPS-induced coagulation in humans. Twenty-four healthy male subjects participated in this randomized, double-blind, placebo-controlled, parallel group study. Volunteers received 2 ng/kg LPS intravenously, followed by a bolus-primed continuous infusion of placebo or lepirudin (Refludan, bolus: 0.1 mg/kg, infusion: 0.1 mg/kg/h for 5 hours) to achieve a 2-fold prolongation of the activated partial thromboplastin time (aPTT). LPS infusion enhanced thrombin activity as evidenced by a 20-fold increase of thrombin-antithrombin complexes (TAT), a 6-fold increase of polymerized soluble fibrin, termed thrombus precursor protein (TpP), and a 4-fold increase in D-dimer. In the lepirudin group, TAT increased only 5-fold, TpP increased by only 50%, and D-dimer only slightly exceeded baseline values (P <.01 versus placebo). Concomitantly, lepirudin also blunted thrombin generation evidenced by an attenuated rise in prothrombin fragment levels (F(1 + 2), P <. 01 versus placebo) and blunted the expression of tissue factor on circulating monocytes. This experimental model proved the anticoagulatory potency of lepirudin in LPS-induced coagulation activation. Results from this trial provide a rationale for a randomized clinical trial on the efficacy of lepirudin in DIC. (Blood. 2000;95:1729-1734) PMID- 10688832 TI - Unique processing pathways within recipient antigen-presenting cells determine IgG immunity against donor platelet MHC antigens. AB - Recipient IgG immunity against leukoreduced donor platelets is dependent on indirect T-cell allorecognition and is suppressed in vivo by inhibitors (aminoguanidine, AMG) of inducible nitric oxide synthase (iNOS). To examine recipient processing pathways of donor platelet antigens, enriched macrophages (antigen-presenting cells [APC]) from BALB/c (H-2(d)) mice were pulsed with allogeneic C57BL/6 (H-2(b)) platelets and transfused weekly into naive BALB/c mice. Platelet-pulsed APC stimulated IgG antidonor antibody production in 45% of recipients by the second transfusion and in 100% by the sixth transfusion; this response was enhanced by pulsing in the presence of interferon-gamma. By the sixth transfusion, high-titer IgG1 (mean titer 4990) and IgG2a (1933) isotypes specific for donor major histocompatibility complex (MHC) class I antigens were detected. Platelet pulsing in the presence of AMG or colchicine significantly inhibited the ability of APC to stimulate IgG alloantibodies; only 50% (P <.005) and 20% (P <.0001) of recipients, respectively, produced antibodies by the sixth transfusion. AMG inhibition was reversed by the addition of L-arginine, the substrate for iNOS. In contrast, pulsing in the presence of chloroquine, the proteasome inhibitory peptide MG115, or Brefeldin A enhanced APC immunity (70 100% of recipients antibody positive by the second transfusion [P <.05]); these agents allowed the pulsed APC to stimulate IgG2a but inhibited IgG1 production and this correlated with a reduction in serum interleukin (IL)-4 levels. The results suggest that for donor platelet antigens to stimulate IgG alloantibodies, recipient APC use the essential generation of nitric oxide and a noncytosolic, pH independent processing pathway, which can be exploited as an effective immunotherapy target to further inhibit alloimmunization against leukoreduced platelets. (Blood. 2000;95:1735-1742) PMID- 10688833 TI - Initiation of antiretroviral therapy during primary HIV-1 infection induces rapid stabilization of the T-cell receptor beta chain repertoire and reduces the level of T-cell oligoclonality. AB - Major T-cell receptor beta chain variable region (TCRBV) repertoire perturbations are temporally associated with the down-regulation of viremia during primary human immunodeficiency virus (HIV) infection and with oligoclonal expansion and clonal exhaustion of HIV-specific cytotoxic T lymphocytes (CTLs). To determine whether initiation of antiretroviral therapy (ART) or highly active antiretroviral therapy (HAART) during primary infection influences the dynamics of T-cell-mediated immune responses, the TCRBV repertoire was analyzed by semiquantitative polymerase chain reaction in serial blood samples obtained from 11 untreated and 11 ART-treated patients. Repertoire variations were evaluated longitudinally. Stabilization of the TCRBV repertoire was more consistently observed in treated as compared with untreated patients. Furthermore, the extent and the rapidity of stabilization were significantly different in treated versus untreated patients. TCRBV repertoire stabilization was positively correlated with the slope of HIV viremia in the treated group, suggesting an association between repertoire stabilization and virologic response to treatment. To test whether stabilization was associated with variations in the clonal complexity of T-cell populations, T-cell receptor (TCR) heteroduplex mobility shift assays (HMAs) were performed on sequential samples from 4 HAART-treated subjects. Densitometric analysis of HMA profiles showed a reduction in the number of TCR clonotypes in most TCRBV families and a significant decrease in the total number of clonotypes following 7 months of HAART. Furthermore, a biphasic decline in HIV-specific but not heterologous CTL clones was observed. This indicates that ART leads to a global reduction of CD8(+) T-cell oligoclonality and significantly modulates the mobilization of HIV-specific CTL during primary infection. (Blood. 2000;95:1743 1751) PMID- 10688834 TI - Inhibition of degranulation and interleukin-6 production in mast cells derived from mice deficient in protein kinase Cbeta. AB - The antigen-mediated activation of mast cells by means of IgE antibodies bound to the cell surface leads to direct interactions between FcepsilonRI receptor cytoplasmic domains and various intracellular proteins. These interactions initiate diverse signal-transduction pathways, and the activation of these pathways results in the immediate release of proinflammatory agents. A delayed response also occurs and includes the release of various cytokines. It is clear that the activation of kinases is a requirement for the exocytosis observed in mast cells. In addition to the tyrosine phosphorylation of the affected system by soluble tyrosine kinases, activity of protein kinase C (PKC) results in serine or threonine phosphorylation of multiple protein substrates. In this study, we found that mast cells derived from PKCbeta-deficient mice produce less interleukin 6 in response to IgE-Ag. The inhibition of exocytosis in the PKCbeta-deficient mast cells occurred whether the stimuli were due to the aggregation of the mast cell surface FcepsilonRI or to the calcium ionophore, ionomycin. However, no significant changes were observed in the proliferative response of the mast cells to interleukin 3 (IL-3) or in their apoptotic rate after IL-3 depletion. (Blood. 2000;95:1752-1757) PMID- 10688835 TI - Induction of resistance to the Abelson inhibitor STI571 in human leukemic cells through gene amplification. AB - The 2-phenylaminopyrimidine derivative STI571 has been shown to selectively inhibit the tyrosine kinase domain of the oncogenic bcr/abl fusion protein. The activity of this inhibitor has been demonstrated so far both in vitro with bcr/abl expressing cells derived from leukemic patients, and in vivo on nude mice inoculated with bcr/abl positive cells. Yet, no information is available on whether leukemic cells can develop resistance to bcr/abl inhibition. The human bcr/abl expressing cell line LAMA84 was cultured with increasing concentrations of STI571. After approximately 6 months of culture, a new cell line was obtained and named LAMA84R. This newly selected cell line showed an IC50 for the STI571 (1.0 microM) 10-fold higher than the IC50 (0.1 microM) of the parental sensitive cell line. Treatment with STI571 was shown to increase both the early and late apoptotic fraction in LAMA84 but not in LAMA84R. The induction of apoptosis in LAMA84 was associated with the activation of caspase 3-like activity, which did not develop in the resistant LAMA84R cell line. LAMA84R cells showed increased levels of bcr/abl protein and mRNA when compared to LAMA84 cells. FISH analysis with BCR- and ABL-specific probes in LAMA84R cells revealed the presence of a marker chromosome containing approximately 13 to 14 copies of the BCR/ABL gene. Thus, overexpression of the Bcr/Abl protein mediated through gene amplification is associated with and probably determines resistance of human leukemic cells to STI571 in vitro. (Blood. 2000;95:1758-1766) PMID- 10688836 TI - MSH2-deficient murine lymphomas harbor insertion/deletion mutations in the transforming growth factor beta receptor type 2 gene and display low not high frequency microsatellite instability. AB - High-frequency microsatellite instability (MSI), defined as more than 20% unstable loci, is an inconsistent finding in hematologic malignancies; consequently, the significance of deficient DNA mismatch repair (MMR) to their pathogenesis has been questioned. To further investigate the relationship between MMR deficiency and genomic instability in hematologic malignancies, this study evaluated MSH2-/- murine lymphomas for insertion/deletion (ID) mutations within the transforming growth factor (TGF)-beta receptor type II (TbetaR-II) gene and MSI at 10 neutral microsatellites. The lymphomas displayed ID mutations within short mononucleotide runs of TbetaR-II at a high frequency, whereas nonmalignant tissue from corresponding animals lacked mutations. Loss of TbetaR-II transcripts and protein was seen in 6 of 7 murine lymphomas harboring acquired TbetaR-II mutations. In the analysis of paired nonmalignant and tumor DNA samples, low frequency but not high-frequency MSI was found. Low-frequency MSI occurred in 8 of 20 lymphomas and 12 displayed microsatellite stability. MSI was even less frequent in nonmalignant tissue as only 3 of 20 samples displayed low-frequency MSI and 17 displayed stability. Evaluation of 20 single cell clones from the MSH2 /- lymphoma cell lines R25 and L15 identified high-frequency MSI in 4 and 2 clones, respectively. The remaining clones showed low-frequency MSI or stability. These findings suggest that acquired TbetaR-II mutations represent important inactivating events in tumor pathogenesis following MSH2 deficiency. Furthermore, for some hematolymphoid malignancies, the evaluation of cancer-associated genes for ID mutations may represent a more sensitive marker of MMR deficiency than evaluation of neutral microsatellites for high-frequency MSI. (Blood. 2000;95:1767-1772) PMID- 10688837 TI - Induction of mitochondrial permeability transition and cytochrome C release in the absence of caspase activation is insufficient for effective apoptosis in human leukemia cells. AB - Induction of mitochondrial permeability transition (MPT) and cytosolic translocation of cytochrome C are considered essential components of the apoptotic pathway. Hence, there is the realization that mitochondrial-specific drugs could have potential for use as chemotherapeutic agents to trigger apoptosis in tumor cells. Recently, we showed that photoproducts of merocyanine 540 (pMC540) induced tumor cell apoptosis. In this study, we focused on identifying mitochondrial-specific compounds from pMC540 and studied their apoptotic potential. One purified fraction, C5, induced a drop in mitochondrial transmembrane potential and cytosolic translocation of cytochrome C in HL60 human leukemia cells. Moreover, the addition of C5 to purified rat liver mitochondria induced MPT as indicated by mitochondrial matrix swelling, which was completely inhibited by cyclosporin A, an inhibitor of the inner-membrane pore. Supernatant of C5-treated mitochondria showed a dose-dependent increase in cytochrome C, which was also inhibited in the presence of cyclosporin A, strongly indicating a direct effect on the inner-membrane pore. Despite the strong mitochondrial reactivity, C5 elicited minimal cytotoxicity (less than 25%) against HL60 leukemia and M14 melanoma cells because of inefficient caspase activation. However, prior exposure to C5 significantly enhanced the apoptotic response to etoposide or the CD95 receptor. Thus, we demonstrate that MPT induction and cytochrome C release by the novel compound C5, in the absence of effective caspase activation, is insufficient for triggering efficient apoptosis in tumor cells. However, when used in combination with known apoptosis inducers, such compounds could enhance the sensitivity of tumor cells to apoptosis. (Blood. 2000;95:1773-1780) PMID- 10688838 TI - Vaccination of patients with chronic myelogenous leukemia with bcr-abl oncogene breakpoint fusion peptides generates specific immune responses. AB - Chronic myelogenous leukemia (CML) presents a unique opportunity to develop therapeutic strategies using vaccination against a truly tumor-specific antigen that is also the oncogenic protein required for neoplasia. CML is characterized by the t(9;22) that results in the bcr-abl fusion oncogene and in the expression of a chimeric protein product p210. Previously we have shown that peptides derived from amino acid sequences crossing the b3a2 fusion breakpoint in p210 elicit class I restricted cytotoxic T lymphocytes and class II responses, respectively, in vitro. Such sequences may thus comprise absolutely tumor specific antigens in a peptide-based vaccine. We evaluated the safety and immunogenicity of a multidose, bcr-abl breakpoint peptide vaccine in 12 adults with chronic-phase CML. Cohorts of 3 patients each received either 50 microg, 150 microg, 500 microg, or 1500 microg total peptide mixed with 100 microg QS-21 as an immunological adjuvant. Delayed-type hypersensitivity (DTH), humoral responses, and unprimed ex vivo autologous proliferation ((3)H-thymidine incorporation) and cytotoxicity (chromium-51 release) responses were measured. All 68 vaccinations were well tolerated without significant adverse effects. In 3 of the 6 patients treated at the 2 highest dose levels of vaccine, peptide specific, T-cell proliferative responses (n = 3) and/or DTH responses (n = 2) were generated that lasted up to 5 months after vaccination. Cytotoxic T lymphocytes have not been identified. In conclusion, a tumor-specific, bcr-abl derived peptide vaccine can be safely administered to patients with chronic-phase CML and can elicit a bcr-abl peptide-specific immune response despite the presence of active disease in these patients and approximately 10(12) leukemia cells. (Blood. 2000;95:1781-1787) PMID- 10688839 TI - FGFR1 is fused to the centrosome-associated protein CEP110 in the 8p12 stem cell myeloproliferative disorder with t(8;9)(p12;q33). AB - The hallmark of the 8p12 stem cell myeloproliferative disorder (MPD) is the disruption of the FGFR1 gene, which encodes a tyrosine kinase receptor for members of the fibroblast growth factor family. FGFR1 can be fused to at least 3 partner genes at chromosomal regions 6q27, 9q33, or 13q12. We report here the cloning of the t(8;9)(p12;q33) and the detection of a novel fusion betweenFGFR1 and the CEP110 gene, which codes for a novel centrosome-associated protein with a unique cell-cycle distribution. CEP110 is widely expressed at various levels in different tissues and is predicted to encode a 994-amino acid coiled-coil protein with 4 consensus leucine zippers [L-X(6)-L-X(6)-L-X(6)-L]. Both reciprocal fusion transcripts are expressed in the patient's cells. The CEP110-FGFR1 fusion protein encodes an aberrant tyrosine kinase of circa 150-kd, which retains most of CEP110 with the leucine zipper motifs and the catalytic domain of FGFR1. Transient expression studies show that the CEP110-FGFR1 protein has a constitutive kinase activity and is located within the cell cytoplasm. (Blood. 2000;95:1788-1796) PMID- 10688840 TI - Molecular analysis of immunoglobulin genes in diffuse large B-cell lymphomas. AB - Diffuse large B-cell lymphoma (DLBCL) is a common type of non-Hodgkin's lymphoma (NHL) that is highly heterogeneous from both clinical and histopathologic viewpoints. The immunoglobulin (Ig) heavy (H) chain variable region genes were examined in 71 patients with untreated primary DLBCL. Fifty-eight potentially functional V(H) genes were detected in 53 DLBCL cases; V(H) genes were nonfunctional in 9 cases and were not detected in an additional 9 cases. The use of V(H) gene families by DLBCL tumors was unbiased without overrepresentation of any particular V(H) gene or gene family. Analysis of Ig mutations in comparison to the most closely related germline gene disclosed mutated V(H) genes in all but 1 DLBCL case. More than 2% difference from the most similar germline sequence was detected in 52 potentially functional and the 8 nonfunctional V(H) gene sequences, whereas less than 2% difference from the germline sequence was observed in 3 V(H) gene isolates. Only 3 V(H) gene isolates were unmutated. No correlation was found between V(H) gene use, mutation level, and International Prognostic Index (IPI) or survival. Six of 8 tested tumors showed evidence of ongoing somatic mutations. Evidence for positive or negative antigen selection pressure was observed in 65% of mutated DLBCL cases. Our findings indicate that the etiology and the driving forces for clonal expansion are heterogeneous, which may explain the well-known clinical and pathologic heterogeneity of DLBCL. (Blood. 2000;95:1797-1803) PMID- 10688841 TI - Integrin alpha(2)beta(1) (VLA-2) is a principal receptor used by neutrophils for locomotion in extravascular tissue. AB - Cell adhesion molecules are critically involved in the multistep process of leukocyte recruitment in inflammation. The specific receptors used by polymorphonuclear leukocytes (PMN) for locomotion in extravascular tissue have as yet not been identified. By means of immunofluorescence flow cytometry and laser scanning confocal microscopy, this study demonstrated that surface expression of the alpha(2)beta(1) (VLA-2) integrin, though absent on blood PMN, is induced in extravasated PMN collected from human skin blister chambers, and rat PMN accumulated in the peritoneal cavity after chemotactic stimulation. Intravital time-lapse videomicroscopy was used to investigate chemoattractant-induced PMN locomotion in the rat mesentery in vivo. Local administration of function blocking monoclonal antibody or peptide recognizing the alpha(2)beta(1) integrin reduced PMN migration velocity in the extravascular tissue by 73% +/- 3% and 70% +/- 10%, respectively (means +/- SD). The distance f-met-leu-phe peptide (fMLP) stimulated human PMN migrated in a collagen gel in vitro was markedly reduced by treatment with anti-alpha(2) mAbs or peptide, whereas no effect was observed with antibodies or peptides recognizing the alpha(4)beta(1) or alpha(5)beta(1) integrins. Further evidence for a critical role of expression of alpha(2)beta(1) integrin in PMN locomotion in extravascular tissue was obtained in the mouse air pouch model of acute inflammation where chemoattractant-induced PMN recruitment was substantially inhibited by local anti-alpha(2) mAb treatment. Thus, expression of alpha(2)beta(1) integrin on extravasated PMN has been identified and a novel role of this receptor in regulating the extravascular phase of leukocyte trafficking in inflammation has been formulated. (Blood. 2000;95:1804 1809) PMID- 10688842 TI - The synthetic chemoattractant Trp-Lys-Tyr-Met-Val-DMet activates neutrophils preferentially through the lipoxin A(4) receptor. AB - A D-methionine-containing peptide, Trp-Lys-Tyr-Met-Val-D-Met-NH(2) (WKYMVm), featuring a unique receptor specificity was investigated with respect to its ability to activate neutrophil effector functions. The peptide was found to be more potent than the N-formylated peptide N-formyl-Met-Leu-Phe (fMLF) at inducing neutrophil chemotaxis, mobilization of neutrophil complement receptor 3 (CR3), and activation of the neutrophil NADPH-oxidase. The fact that binding of fML[(3)H]F was inhibited by both fMLF and WKYMVm suggests that N-formyl peptide receptor (FPR) is shared by these peptides. However, the neutrophil response induced by the WKYMVm peptide was insensitive to the fMLF antagonists, cyclosporin H, and Boc-FLFLF that specifically block the function of the FPR. These results suggest that even though WKYMVm may bind FPR the cells are activated preferentially through a receptor distinct from the FPR. Using transfected HL-60 cells expressing either the FPR or its neutrophil homologue FPRL1, also referred to as LXA(4)R because it has been shown to bind lipoxin A(4), we show that WKYMVm is about 300-fold more active at mobilizing intracellular calcium through FPRL1 than through FPR. The WKYMVm activates FPRL1 expressing cells in a cyclosporin H-independent manner with an EC(50 )of around 75 pmol/L, whereas it activates FPR-expressing cells with an EC(50 )of around 25 nmol/L. The observation that exudated cells are primed in their response to WKYMVm suggests that FPRL1/LXA(4)R like FPR is stored in mobilizable organelles. (Blood. 2000;95:1810-1818) PMID- 10688843 TI - A study of the coregulation and tissue specificity of XG and MIC2 gene expression in eukaryotic cells. AB - CD99, the product of the MIC2 gene, exhibits an erythroid-specific quantitative polymorphism coregulated with the polymorphism of the XG blood group gene. As a preliminary study of this phenomenon, human XG and CD99 recombinant proteins were expressed in murine RAG cells and analyzed by flow cytometry. Both proteins were expressed independently and at a similar level in single and double transfectants. Immunoprecipitation and Western blot analysis, using the murine monoclonal antibodies NBL-1 and 12E7, revealed species of 26 kd (XG) and 32 kd (CD99), respectively. A putative 28-kd intracellular precursor of CD99 was also detected, as was a 26-kd species after neuraminidase treatment of CD99-expressing cells. No evidence of association or complex formation between XG and CD99 proteins could be proven, either on transfected RAG cells or on human erythrocytes. These results were confirmed using somatic hybrids between single transfectants. These findings suggest that the phenotypic relationship between XG and CD99 is mostly regulated at the transcriptional level, but they do not formally exclude some posttranscriptional effect. Studies on the tissue specificity of XG expression showed that surface expression of the XG protein could not be restored in somatic hybrids between B-lymphoblastoid cell lines from Xg(a+) persons and fibroblasts (RAG) or erythroid (MEL) cells. RT-PCR analysis of the transcripts revealed the existence of an XG mRNA in each cell line, suggesting that the tissue-specific regulation of cell surface XG expression occurs either at a quantitative transcriptional level or is a posttranscriptional event. By Northern blot analysis, XG transcripts were detected in erythroid tissues and several nonerythroid tissues. (Blood. 2000;95:1819-1826) PMID- 10688844 TI - Fetal expression of a human Agamma globin transgene rescues globin chain imbalance but not hemolysis in EKLF null mouse embryos. AB - Mice lacking the erythroid Kruppel-like factor (EKLF) die in utero at embryonic day 15 (E15) from severe anemia. EKLF(-/-) embryos display a marked deficit in beta-globin gene expression. To test whether beta-globin deficiency was solely responsible for the anemia and intrauterine death, we corrected the globin chain imbalance in EKLF(-/-) embryos by breeding with a strain of mice that express high levels of human gamma-globin. Despite efficient production of hybrid malpha(2)-hgamma(2) hemoglobin in the fetal livers of EKLF(-/-) animals, hemolysis was not corrected and survival was not prolonged. We concluded that deficiency of nonglobin EKLF target genes is a major contributor to the definitive red blood cell abnormalities and prenatal death in EKLF(-/-) embryos. These results suggest that strategies designed to antagonize EKLF function in adults with hemoglobinopathy, in an attempt to reactivate gamma-globin gene expression, may adversely affect other essential aspects of red blood cell physiology. (Blood. 2000;95:1827-1833) PMID- 10688845 TI - Elliptocytosis in patients with C-terminal domain mutations of protein 4.1 correlates with encoded messenger RNA levels rather than with alterations in primary protein structure. AB - Early biochemical studies defined 4 functional domains of the erythroid protein 4.1 (4.1R). From amino-terminal to carboxy-terminal, these are 30 kd, 16 kd, 10 kd, and 22/24 kd in size. Although the functional properties of both the 30-kd and the 10-kd domain have been demonstrated in red cells, no functional activities have been assigned to either the 16-kd or the 22/24-kd domain in these cells. We here describe new mutations in the sequence encoding the C-terminal 22/24-kd domain that are associated with hereditary elliptocytosis. An unusually mild phenotype observed in heterozygous and homozygous members of 1 family suggested heterogeneity in the pattern of expression of 4.1R deficiency. Using a variety of protein and messenger RNA (mRNA) quantification strategies, we showed that, regardless of the alteration in the C-terminal primary sequence, when the protein is produced, it assembles at the cell membrane. In addition, we found that alterations in red cell morphologic features and membrane function correlate with the amount of membrane-associated protein-and therefore with the amount of mRNA accumulated-rather than with the primary structure of the variant proteins. These data suggest that an intact sequence at exons 19 through 21 encoding part of the C-terminal 22/24-kd region is not required for proper protein 4.1R assembly in mature red cells. (Blood. 2000;95:1834-1841) PMID- 10688846 TI - Volume control in sickle cells is facilitated by the novel anion conductance inhibitor NS1652. AB - A low cation conductance and a high anion conductance are characteristic of normal erythrocytes. In sickle cell anemia, the polymerization of hemoglobin S (HbS) under conditions of low oxygen tension is preceded by an increase in cation conductance. This increase in conductance is mediated in part through Ca(++) activated K(+) channels. A net efflux of potassium chloride (KCl) leads to a decrease in intracellular volume, which in turn increases the rate of HbS polymerization. Treatments minimizing the passive transport of ions and solvent to prevent such volume depletion might include inhibitors targeting either the Ca(++)-activated K(+) channel or the anion conductance. NS1652 is an anion conductance inhibitor that has recently been developed. In vitro application of this compound lowers the net KCl loss from deoxygenated sickle cells from about 12 mmol/L cells/h to about 4 mmol/L cells/h, a value similar to that observed in oxygenated cells. Experiments performed in mice demonstrate that NS1652 is well tolerated and decreases red cell anion conductance in vivo. (Blood. 2000;95:1842 1848) PMID- 10688847 TI - Single amino acid substitution in human platelet glycoprotein Ibbeta is responsible for the formation of the platelet-specific alloantigen Iy(a). AB - We recently described a new low-frequency platelet alloantigen on the human platelet glycoprotein (GP) Ib-IX complex, termed Iy(a), which was implicated in a severe case of neonatal alloimmune thrombocytopenia. Immunoprecipitation studies with trypsin-treated platelets indicated that the Iy(a) alloantigenic determinants are formed by the membrane-associated remnant moiety of GP Ibalpha (GP Ibalpha(r)) together with GP Ibbeta and GP IX. To elucidate the molecular basis underlying the Iy(a) alloantigen, we amplified GPIbalpha(r), GPIbbeta, and GPIX genes by polymerase chain reaction (PCR). Nucleotide-sequence analysis of these 3 genes showed a G to A transition at position 141 on GPIbbeta gene in a subject positive for Iy(a). This transition resulted in a Gly(15)Glu dimorphism on the N-terminal domain of GPIbbeta. This finding was confirmed by genotyping analysis of 6 Iy(a)-positive subjects by restriction fragment length polymorphism (RFLP) studies using NarI endonuclease. In 300 randomly selected healthy blood donors, one Iy(a)-positive individual was found. Phenotypes determined by monoclonal antibody-specific immobilization of platelet antigens assay and genotypes determined by RFLP were identical in this population. Analysis of Iy(a) positive platelets showed that the point mutation affected neither the degree of surface expression nor the function of the GP Ibalpha-GP Ibbeta-IX complex on the platelet surface. Transient expression of the GP Ib-IX complex in CHO cells using wild-type GP Ibbeta (Gly(15)) or mutant GP Ibbeta (Glu(15)) allowed us to demonstrate that this single amino acid substitution is sufficient to induce Iy(a) epitope(s). (Blood. 2000;95:1849-1855) PMID- 10688848 TI - Vasoactive side effects of intravenous immunoglobulin preparations in a rat model and their treatment with recombinant platelet-activating factor acetylhydrolase. AB - Previously, we observed in a rat model that intravenous administration of intramuscular immunoglobulin preparations induced a long-lasting hypotension, which appeared to be associated with the presence of IgG polymers and dimers in the preparations, but unrelated to complement activation. We found evidence that this hypotensive response is mediated by platelet-activating factor (PAF) produced by macrophages. In this study, we compared the vasoactive effects of 16 intravenous immunoglobulin (IVIG) products from 10 different manufacturers, in anesthetized rats. Eight of the IVIG preparations showed no hypotensive effects (less than 15% decrease), whereas the other 8 had relatively strong effects (15% 50% decrease). The hypotensive effects correlated with the IgG dimer content of the preparations. Pretreatment of the rats with recombinant PAF acetylhydrolase completely prevented the hypotensive reaction on IVIG infusion, and administration after the onset of hypotension resulted in normalization of the blood pressure. We also observed PAF production on in vitro incubation of human neutrophils with IVIG, which could be blocked by anti-Fcgamma receptor antibodies. This indicates that induction of PAF generation may also occur in a human system. Our findings support the hypothesis that the clinical side effects of IVIG in patients may be caused by macrophage and neutrophil activation through interaction of IgG dimers with Fcgamma receptors. Because phagocyte activation may also lead to the release of other inflammatory mediators, recombinant PAF acetylhydrolase (rPAF-AH) provides a useful tool to determine whether PAF plays a role in the clinical side effects of IVIG. If so, rPAF-AH can be used for the treatment of those adverse reactions. (Blood. 2000;95:1856-1861) PMID- 10688850 TI - Germline CDKN2A mutation implicated in predisposition to multiple myeloma. AB - Germline mutations of the CDKN2A (p16(INK4A)) tumor suppressor gene predispose patients to melanoma and pancreatic carcinoma. In contrast, mutations of the murine CDKN2A gene predispose BALB/c mice to pristane-induced plasmacytoma. We describe here a family in which a germline mutation of CDKN2A is present in 4 individuals who developed melanoma as well as in a fifth family member who is suffering from multiple myeloma. To determine whether the CDKN2A mutation predisposed the myeloma patient to her disease, we carried out loss of heterozygosity studies on sorted bone marrow from this individual and observed loss of the wild type CDKN2A allele in the malignant plasma cells. We suggest that germline mutations of CDKN2A may predispose individuals to a wider variety of malignancy than has been hitherto reported, but that the expression of these cancers may depend heavily on the genetic background of the patient. (Blood. 2000;95:1869-1871) PMID- 10688849 TI - Treatment of intractable autoimmune diseases in MRL/lpr mice using a new strategy for allogeneic bone marrow transplantation. AB - A new bone marrow transplantation (BMT) method for treating severe autoimmune diseases in chimeric resistant MRL/lpr mice is presented. The method consists of fractionated irradiation (5.5 Gy x 2), followed by portal venous (PV) injection of whole bone marrow cells (BMCs) from allogeneic normal C57BL/6 (B6) mice and intravenous (IV) injection of whole B6 BMCs 5 days after the PV injection (abbreviated as 5.5 Gy x 2 + PV + IV). All recipients survived more than 1 year after this treatment (more than 64 weeks after birth). Abnormal T cells (Thy1.2(+)/B220(+)/CD3(+)/CD4(-)/CD8(-)) present in MRL/lpr mice before the treatment disappear, and hematolymphoid cells are reconstituted with donor derived cells. The treated mice are free from autoimmune diseases. Levels of autoantibodies (IgG/IgM anti-ssDNA antibodies and IgG/IgM rheumatoid factors) decrease to normal levels. Successful cooperation is achieved among T cells, B cells, and antigen-presenting cells (APCs) of the treated MRL/lpr mice when evaluated by in vitro anti-SRBC responses. Newly developed T cells are tolerant to both donor (B6)-type and host (MRL/lpr)-type major histocompatibility complex (MHC) determinants. These findings clearly indicate that severe autoimmune diseases in MRL/lpr mice are completely ameliorated by the treatment without recourse to immunosuppressants, and that the treated MRL/lpr mice show normal immune functions, strongly suggesting that this strategy would be applicable to humans. (Blood. 2000;95:1862-1868) PMID- 10688851 TI - Progenitor cell mobilization by granulocyte colony-stimulating factor controlled by loci on chromosomes 2 and 11. AB - Granulocyte colony-stimulating factor (G-CSF) can effectively mobilize hematopoietic stem and progenitor cells from bone marrow into blood, thereby allowing peripheral blood stem cells (PBSCs) to be used for transplantation. The efficiency of PBSC mobilization response to G-CSF is a multigene trait. DBA/2 (high-responder) and C57BL/6 (low-responder) mice were used for a genetic analysis of G-CSF-induced progenitor release. Significant linkages were found on chromosome 2 by analyzing segregation distortion among the high responders of 500 backcross mice and on chromosome 11 by using the quantitative trait locus analysis of 26 strains of BXD recombinant inbred mice. (Blood. 2000;95:1872-1874) PMID- 10688852 TI - Carcinogenesis, 1980 PMID- 10688853 TI - Overview of carcinogenesis: past, present and future. AB - In the foregoing articles, the editors of Carcinogenesis have identified the major themes of current carcinogenesis research and assembled an outstanding group of authors to review these areas. I have been asked to provide a historical overview of past accomplishments and describe how these contributed to the broader efforts to overcome the burden of human cancer. My assignment also included a look into the future. As scientists we formulate hypotheses that attempt to predict the future. Occasionally we are successful. The pioneers of carcinogenesis research were remarkably successful in predicting the future. Armed with primitive technology relative to today, these scientists studied the biology of carcinogenesis and conceptualized a framework for cancer pathogenesis that virtually everyone working in cancer research follows today. The current generation has been charged with filling in the details. In the details lay the future. Together with past accomplishments, these emerging details form a remarkable picture of progress in understanding and application, creating realistic and imminent promise to achieve victory in the fight against cancer. PMID- 10688854 TI - Metabolism of chemical carcinogens. AB - The transformation of chemicals is important in carcinogenesis, both in bioactivation and detoxification. Major advances in the past 20 years include appreciation of the migration of reactive electrophiles, the ability of Phase II conjugating enzymes to activate chemicals, understanding of the human enzymes, the realization that DNA modification can result from endogenous chemicals, and the demonstration that cancers can result from the metabolism of chemicals to non covalently bound products. Pathways of transformation in which major insight was gained during the past 20 years include nitropolycyclic hydrocarbons, polycyclic hydrocarbons and their diols, vinyl halides and dihaloalkanes. Advances in analytical methods and recombinant DNA technology contributed greatly to the study of metabolism of chemical carcinogens. Major advances have been made in the assignment of roles of individual enzymes in reactions. The knowledge developed in this field has contributed to growth in the areas of chemoprevention, molecular epidemiology and species comparisons of risk. Some of the areas in which future development relevant to carcinogen metabolism is expected involve pathways of transformation of certain chemicals, regulation of genes coding for many of the enzymes under consideration and genomics. PMID- 10688855 TI - Carcinogen macromolecular adducts and their measurement. AB - Damage to DNA induced by carcinogenic chemicals reflects exposure and is directly related to tumor formation, whereas modification of protein provides relatively precise dosimetry for stable adducts of proteins with a known half-life. Sophisticated methods for the detection and quantitation of DNA and protein adducts have been developed during the last approximately 25 years. For DNA adducts the most widely used methods include electrochemical detection, mass spectrometry, fluorescence and phosphorescence spectroscopy, immunoassays and immunohistochemistry and (32)P-post-labeling. Detection limits for quantitative assays are typically in the range of 1 adduct in 10(7) or 10(9) nucleotides. However, accelerator mass spectrometry, which is highly sophisticated but less accessible, has a detection limit of approximately 1 adduct in 10(12) nucleotides. Methods for the determination of protein adducts include immunoassay and a variety of elegant high-resolution mass spectrometry approaches. The detection limit of approximately 0.1 fmol for protein adducts, is based primarily on method specificity and the availability of large quantities of sample material. Using these highly sensitive methods a major achievement has been the biomonitoring of chemically exposed human populations. Validation of macromolecular adduct formation in humans has been predicated on studies in animal models. Adduct formation in humans appears to be indicative of molecular dosimetry and suggestive of increased human cancer risk. However, despite the large body of literature documenting DNA and protein adduct molecular dosimetry for many carcinogen exposures, the relationship between adduct formation and human cancer risk has been defined for only a few carcinogens. Thus, elucidation of this association remains a compelling challenge. For the future, integration of DNA and protein adduct measurements together with documentation of correlative and subsequent events, and host susceptibility factors, within the context of valid molecular epidemiologic study designs, will further our understanding of human disease mechanisms. PMID- 10688856 TI - Oxyradicals and DNA damage. AB - A major development of carcinogenesis research in the past 20 years has been the discovery of significant levels of DNA damage arising from endogenous cellular sources. Dramatic improvements in analytical chemistry have provided sensitive and specific methodology for identification and quantitation of DNA adducts. Application of these techniques to the analysis of nuclear DNA from human tissues has debunked the notion that the human genome is pristine in the absence of exposure to environmental carcinogens. Much endogenous DNA damage arises from intermediates of oxygen reduction that either attack the bases or the deoxyribosyl backbone of DNA. Alternatively, oxygen radicals can attack other cellular components such as lipids to generate reactive intermediates that couple to DNA bases. Endogenous DNA lesions are genotoxic and induce mutations that are commonly observed in mutated oncogenes and tumor suppressor genes. Their mutagenicity is mitigated by repair via base excision and nucleotide excision pathways. The levels of oxidative DNA damage reported in many human tissues or in animal models of carcinogenesis exceed the levels of lesions induced by exposure to exogenous carcinogenic compounds. Thus, it seems likely that oxidative DNA damage is important in the etiology of many human cancers. This review highlights some of the major accomplishments in the study of oxidative DNA damage and its role in carcinogenesis. It also identifies controversies that need to be resolved. Unraveling the contributions to tumorigenesis of DNA damage from endogenous and exogenous sources represents a major challenge for the future. PMID- 10688857 TI - Carcinogenesis in mouse and human cells: parallels and paradoxes. AB - It has been known since the last century that genetic changes are important in carcinogenesis [Boveri,T. (1914) Zur Frage der Erstehung Maligner Tumoren. Gustav Fischer, Jena]. Observations of tumor cells growing in tissue culture led to the prediction, even before the true nature of the genetic material was known, that alterations at the chromosomal level were critically involved in the process of neoplastic development. The past 20 years have seen the transition of carcinogenesis studies from the purely observational to the molecular genetic level. Although much more needs to be done, it is nevertheless gratifying to be able to piece together the sequence of events from carcinogen exposure, metabolism of the carcinogen to the activated form, formation of specific carcinogen-DNA adducts, misrepair leading to the fixation of mutations in particular target genes, and the resulting selective outgrowth of neoplastic cells. The nature of many of these steps has been clarified only in the relatively recent past, and only for a small number of specific target genes, but the fact that we can say with confidence that such processes occur and are causal changes in tumorigenesis represents a tremendous advance over the situation pertaining 20 years ago. The purpose of this review is to summarize the advances over this time period in our understanding of some of the genetic alterations that contribute to neoplasia, with particular emphasis on chemical carcinogenesis in rodents and the parallels with transformation of human cells. PMID- 10688858 TI - Significance of multiple mutations in cancer. AB - There is increasing evidence that in eukaryotic cells, DNA undergoes continuous damage, repair and resynthesis. A homeostatic equilibrium exists in which extensive DNA damage is counterbalanced by multiple pathways for DNA repair. In normal cells, most DNA damage is repaired without error. However, in tumor cells this equilibrium may be skewed, resulting in the accumulation of multiple mutations. Among genes mutated are those that function in guaranteeing the stability of the genome. Loss of this stability results in a mutator phenotype. Evidence for a mutator phenotype in human cancers includes the frequent occurrence of gene amplification, microsatellite instability, chromosomal aberrations and aneuploidy. Current experiments have centered on two mechanisms for the generation of genomic instability, one focused on mutations in mismatch repair genes resulting in microsatellite instability, and one focused on mutations in genes that are required for chromosomal segregation resulting in chromosomal aberrations. This dichotomy may reflect only the ease by which these manifestations can be identified. Underlying both pathways may be a more general phenomenon involving the selection for mutator genes during tumor progression. During carcinogenesis there is selection for cells harboring mutations that can overcome adverse conditions that limit tumor growth. These mutations are produced by direct DNA damage as well as secondarily as a result of mutations in genes that cause a mutator phenotype. Thus, as tumor progression selects for cells with specific mutations, it also selects for cancer cells harboring mutations in genes that normally function in maintaining genetic instability. PMID- 10688859 TI - Nutrition and dietary carcinogens. AB - Three major factors for human carcinogenesis are (i) cigarette smoking, (ii) infection and inflammation and (iii) nutrition and dietary factors. Nutrition and dietary factors include two categories, namely genotoxic agents and constituents including tumor promotion-associated phenomena. This article first describes the genotoxic agents as microcomponents. These are mutagens/carcinogens in cooked food, fungal products, plant and mushroom substance, and nitrite-related materials, polycyclic aromatic hydrocarbons and oxidative agents. Emphasis has been given to heterocyclic amines (HCAs) to which humans are continuously exposed in an ordinary lifestyle. HCAs in food are mainly produced from creatin(in)e, sugar and from amino acids in meat (upon heating). They are imidazoquinoline and imidazoquinoxaline derivatives and phenylimidazopyridine. HCAs are pluripotent in producing cancers in various organs including breast, colon and prostate. Discussion is also given to plant flavonoids which are mutagenic but not carcinogenic. As a macrocomponent, overintake of total calories, fat and sodium chloride is discussed from the viewpoint of the increase of genetic alterations in tissues and of tumor promotion-associated issues. Studies of nutrition and dietary condition will eventually lead us to cancer prevention, namely delay of onset of cancer to the late phase of human life, which is called 'natural-end cancer' (Tenju-gann). PMID- 10688860 TI - Radiation carcinogenesis. AB - Research on radiation carcinogenesis during the past 2 decades has focused on cellular and molecular mechanisms for the effects of radiation in mammalian cells. This paper will review several of these areas of research, as they may relate specifically to the induction of cancer by ionizing radiation. Knowledge of the critical DNA damage of biologic importance, and how this damage is repaired, will be discussed in relation to its role in the induction of mutations by radiation. The search for the initiating event in radiation carcinogenesis, as well as other genetic events that may be involved, is discussed in terms of the possible role of the activation of oncogenes or tumor suppressor genes and the loss of cell-cycle checkpoints. Finally, evidence will be described indicating that important genetic consequences of radiation may arise in cells that in themselves receive no direct nuclear irradiation. It has been shown that radiation can, by itself, induce a type of genomic instability in cells, which enhances the rate at which mutations and other genetic changes arise in the descendants of the irradiated cell after many generations of replication. Preliminary evidence has been presented that irradiation targeted to the cytoplasm yields a significant increase in the frequency of mutations. Finally, genetic events including the induction of mutations and changes in gene expression may occur in neighboring cells that receive no direct radiation exposure at all. This 'bystander effect' involves gap junction mediated cell-cell communication, and activation of the p53 damage response pathway. The possible role of these phenomena in radiation carcinogenesis is discussed. PMID- 10688861 TI - Viral carcinogenesis: revelation of molecular mechanisms and etiology of human disease. AB - The RNA and DNA tumor viruses have made fundamental contributions to two major areas of cancer research. Viruses were vital, first, to the discovery and analysis of cellular growth control pathways and the synthesis of current concepts of cancer biology and, second, to the recognition of the etiology of some human cancers. Transforming retroviruses carry oncogenes derived from cellular genes that are involved in mitogenic signalling and growth control. DNA tumor viruses encode oncogenes of viral origin that are essential for viral replication and cell transformation; viral oncoproteins complex with cellular proteins to stimulate cell cycle progression and led to the discovery of tumor suppressors. Viral systems support the concept that cancer development occurs by the accumulation of multiple cooperating events. Viruses are now accepted as bona fide etiologic factors of human cancer; these include hepatitis B virus, Epstein Barr virus, human papillomaviruses, human T-cell leukemia virus type I and hepatitis C virus, plus several candidate human cancer viruses. It is estimated that 15% of all human tumors worldwide are caused by viruses. The infectious nature of viruses distinguishes them from all other cancer-causing factors; tumor viruses establish long-term persistent infections in humans, with cancer an accidental side effect of viral replication strategies. Viruses are usually not complete carcinogens, and the known human cancer viruses display different roles in transformation. Many years may pass between initial infection and tumor appearance and most infected individuals do not develop cancer, although immunocompromised individuals are at elevated risk of viral-associated cancers. Variable factors that influence viral carcinogenesis are reviewed, including possible synergy between viruses and environmental cofactors. The difficulties in establishing an etiologic role for a virus in human cancer are discussed, as well as the different approaches that proved viral links to cancer. Future directions for tumor virus studies are considered. PMID- 10688862 TI - Hormonal carcinogenesis. AB - Hormone-related cancers, namely breast, endometrium, ovary, prostate, testis, thyroid and osteosarcoma, share a unique mechanism of carcinogenesis. Endogenous and exogenous hormones drive cell proliferation, and thus the opportunity for the accumulation of random genetic errors. The emergence of a malignant phenotype depends on a series of somatic mutations that occur during cell division, but the specific genes involved in progression of hormone-related cancers are currently unknown. In this review, the epidemiology of endometrial cancer and breast cancer are used to illustrate the paradigms of hormonal carcinogenesis. Then, new strategies for early detection and prevention of hormonal carcinogenesis are discussed. This includes developing polygenic models of cancer predisposition and the further development of safe and effective chemopreventives that block target sequence activity. We developed polygenic models for breast and prostate cancer after hypothesizing that functionally relevant sequence variants in genes involved in steroid hormone metabolism and transport would act together, and also interact with well-known hormonally related risk factors, to define a high-risk profile for cancer. A combination of genes each with minor variation in expressed activity could provide a degree of separation of risk that would be clinically useful as they could yield a large cumulative difference after several decades. The genes included in the breast cancer model are the 17beta-hydroxysteroid dehydrogenase 1 (HSD17B1) gene, the cytochrome P459c17alpha (CYP17) gene, the aromatase (CYP19) gene, and the estrogen receptor alpha (ER) gene. The prostate cancer model includes the androgen receptor gene (AR), steroid 5alpha-reductase type II (SRD5A2), CYP17 and the 3beta hydroxysteroid dehydrogenase (HSD3B2) gene. We present data from our multi-ethnic cohort to support these models. PMID- 10688863 TI - Manipulating the germline: its impact on the study of carcinogenesis. AB - Over the past two decades, the mouse has established itself as the primary organism in which to investigate the fundamental mechanisms of carcinogenesis and to model human neoplasia. The principal reason underlying such dominance almost certainly arises out of our ever increasing ability to manipulate the murine germline. Over the past 20 years we have moved from a position where animal models arose either spontaneously or were generated through exposure to carcinogen to a position in which it is possible to create and study precise mutations of choice. The most recent advances in inducible and conditional technologies now open the possibility for both temporal and tissue-specific gene manipulation. Each of these technological breakthroughs has facilitated significant steps forward in our understanding of the genetic basis of tumorigenesis. This review will highlight some of the major advances in the production and use of murine models of neoplasia over the last two decades. PMID- 10688864 TI - Genome changes and gene expression in human solid tumors. AB - Genome-wide analysis techniques such as chromosome painting, comparative genomic hybridization, representational difference analysis, restriction landmark genome scanning and high-throughput analysis of LOH are now accelerating high-resolution genome aberration localization in human tumors. These techniques are complemented by procedures for detection of differentially expressed genes such as differential display, nucleic acid subtraction, serial analysis of gene expression and expression microarray analysis. These efforts are enabled by work from the human genome program in physical map development, cDNA library production/sequencing and in genome sequencing. This review covers several commonly used large-scale genome and gene expression analysis techniques, outlines genomic approaches to gene discovery and summarizes information that has come from large-scale analyses of human solid tumors. PMID- 10688865 TI - Nucleotide excision repair and human syndromes. AB - DNA damage is implicated in cancer and aging, and several DNA repair mechanisms exist that safeguard the genome from these deleterious consequences. Nucleotide excision repair (NER) removes a wide diversity of lesions, the main of which include UV-induced lesions, bulky chemical adducts and some forms of oxidative damage. The NER process involves the action of at least 30 proteins in a 'cut-and paste'-like mechanism. The consequences of a defect in one of the NER proteins are apparent from three rare recessive syndromes: xeroderma pigmentosum (XP), Cockayne syndrome (CS) and the photosensitive form of the brittle hair disorder trichothiodystrophy (TTD). Sun-sensitive skin is associated with skin cancer predisposition in the case of XP, but remarkably not in CS and TTD. Moreover, the spectrum of clinical symptoms differs considerably between the three syndromes. CS and TTD patients exhibit a spectrum of neurodevelopmental abnormalities and, in addition, TTD is associated with ichthyosis and brittle hair. These typical CS and TTD abnormalities are difficult to comprehend as a consequence of defective NER. This review briefly describes the biochemistry of the NER process, summarizes the clinical features of the NER disorders and speculates on the molecular basis underlying these pleitropic syndromes. PMID- 10688866 TI - DNA methylation: past, present and future directions. AB - DNA methylation, or the covalent addition of a methyl group to cytosine within the context of the CpG dinucleotide, has profound effects on the mammalian genome. These effects include transcriptional repression via inhibition of transcription factor binding or the recruitment of methyl-binding proteins and their associated chromatin remodeling factors, X chromosome inactivation, imprinting and the suppression of parasitic DNA sequences. DNA methylation is also essential for proper embryonic development; however, its presence can add an additional burden to the genome. Normal methylation patterns are frequently disrupted in tumor cells with global hypomethylation accompanying region-specific hypermethylation. When these hypermethylation events occur within the promoter of a tumor suppressor gene they will silence the gene and provide the cell with a growth advantage in a manner akin to deletions or mutations. Recent work indicating that DNA methylation is an important player in both DNA repair and genome stability as well as the discovery of a new family of DNA methyltransferases makes now a very exciting period for the methylation field. This review will highlight the major findings in the methylation field over the past 20 years then summarize the most important and interesting future directions the field is likely to take in the next millennium. PMID- 10688867 TI - Stem cells: the intestinal stem cell as a paradigm. AB - Stem cell research provides a foundation for therapeutic advancement in oncology, clinical genetics and a diverse array of degenerative disorders. For example, the elucidation of pathways governing proliferative regulation and differentiation within cellular systems will result in medical strategies aimed at the root cause of cancer. At present the characterization of reliable stem cell markers is the immediate aim in this particular field. Over the past 30 years investigators have determined many of the physical and functional properties of stem cells through careful and imaginative experimentation. Intestinal stem cells reside at the crypt base and give rise to all cell types found within the crypt. They readily undergo altruistic apoptosis in response to toxic stimuli although their progeny are hardier and will regain stem cell function to repopulate the tissue compartment, giving rise to the concept of a proliferative hierarchy. Contention exists when deciding whether the full complement of cells within a crypt is derived from either a single or multiple stems. Evidence has also arisen to challenge the long held view that colorectal tumours arise from a single mutated stem cell, as early adenomas from a human XO/XY mosaic contained distinct clones. Mechanisms governing the stem cell cycle and subsequent proliferative activity largely remain obscure. The adenomatous polyposis coli gene product has, however, been shown to promote the degradation of beta-catenin, an enhancer of cell proliferation, thereby downregulating this activity in healthy individuals. PMID- 10688868 TI - The role of senescence and immortalization in carcinogenesis. AB - Normal somatic cells are able to divide only a limited number of times before they become senescent. The occurrence of intratumoral cell death and the need for clonal evolution mean that many more cell divisions are required for tumorigenesis than is possible unless cells breach the senescence proliferation barrier and become immortalized. Senescence may therefore be a major tumor suppressor mechanism. During the past decade the study of senescence and immortalization has entered the mainstream of cancer research. A major reason for the current interest in this subject is the observation that most cancers have an activated telomere maintenance mechanism, a marker of immortalization. It has also been found that some of the most common genetic changes known to occur in cancer have a key role in the immortalization process. PMID- 10688869 TI - Apoptosis in cancer. AB - In the last decade, basic cancer research has produced remarkable advances in our understanding of cancer biology and cancer genetics. Among the most important of these advances is the realization that apoptosis and the genes that control it have a profound effect on the malignant phenotype. For example, it is now clear that some oncogenic mutations disrupt apoptosis, leading to tumor initiation, progression or metastasis. Conversely, compelling evidence indicates that other oncogenic changes promote apoptosis, thereby producing selective pressure to override apoptosis during multistage carcinogenesis. Finally, it is now well documented that most cytotoxic anticancer agents induce apoptosis, raising the intriguing possibility that defects in apoptotic programs contribute to treatment failure. Because the same mutations that suppress apoptosis during tumor development also reduce treatment sensitivity, apoptosis provides a conceptual framework to link cancer genetics with cancer therapy. An intense research effort is uncovering the underlying mechanisms of apoptosis such that, in the next decade, one envisions that this information will produce new strategies to exploit apoptosis for therapeutic benefit. PMID- 10688870 TI - Tumor progression and metastasis. AB - It is now widely accepted that cancer is attributed to the accumulation of genetic alterations in cells. Thus, to understand the molecular mechanisms of cancer metastasis, it is indispensable to identify the genes whose alterations accumulate during cancer progression as well as the genes whose expression is responsible for the acquisition of metastatic potential in cancer cells. Molecular analyses of cancer cells in various stages of progression have revealed that alterations in tumor suppressor genes and oncogenes accumulate during tumor progression and correlate with the clinical aggressiveness of cancer. Comparative analyses of gene expression profiles between metastatic and non-metastatic cells have revealed that various genes are differentially expressed in association with the metastatic potential of cancer cells. A number of genes have been also identified as having functions in inducing or suppressing metastasis in experimental models. However, the association between causative genetic alterations and resulting phenotypic alterations with respect to the metastatic potential of cancer cells is not fully understood. Therefore, elucidation of genotype-phenotype correlation will be required to further understand a complex process of metastasis. Here, I review the progress on molecular studies of tumor progression and metastasis of the past 20 years and discuss the future direction in this field of science. PMID- 10688871 TI - Tumor angiogenesis: past, present and the near future. AB - The concept of treating solid tumors by inhibiting tumor angiogenesis was first articulated almost 30 years ago. For the next 10 years it attracted little scientific interest. This situation changed, relatively slowly, over the succeeding decade with the discovery of the first pro-angiogenic molecules such as basic fibroblast growth factor and vascular endothelial growth factor (VEGF), and the development of methods of successfully growing vascular endothelial cells in culture as well as in vivo assays of angiogenesis. However, the 1990s have witnessed a striking change in both attitude and interest in tumor angiogenesis and anti-angiogenic drug development, to the point where a remarkably diverse group of over 24 such drugs is currently undergoing evaluation in phase I, II or III clinical trials. In this review I will discuss the many reasons for this. These features, together with other recent discoveries have created intense interest in initiating and expanding anti-angiogenic drug discovery programs in both academia and industry, and the testing of such newly developed drugs, either alone, or in various combinations with conventional cytotoxic therapeutics. However, significant problems remain in the clinical application of angiogenesis inhibitors such as the need for surrogate markers to monitor the effects of such drugs when they do not cause tumor regressions, and the design of clinical trials. Also of concern is that the expected need to use anti-angiogenic drugs chronically will lead to delayed toxic side effects in humans, which do not appear in rodents, especially in short-term studies. PMID- 10688872 TI - Molecular epidemiology: recent advances and future directions. AB - In 1982 we proposed the concept and a framework for implementing molecular cancer epidemiology. Here, we review progress during the past 17 years in validating and applying this approach to cancer prevention. There have been major advances, notably in the understanding of environment-susceptibility interactions in human cancer. However, a review of major findings to date reveals several urgent research needs to keep pace with the rapid evolution in knowledge of mechanisms in carcinogenesis. Although much valuable progress continues to be made in the study of carcinogens that cause direct DNA damage and are mutagenic, exogenous and endogenous carcinogens can also act by altering gene expression, cell proliferation and differentiation. The mechanisms include aberrant DNA methylation, oxidative damage, effects on metabolism of nitrogen oxide and nitrites, activation of receptors and transcription factors, cyclins and other cell cycle proteins. Sensitive, validated biomarkers are needed to detect these mechanisms in small numbers of cells, tissues or fluids. There is also increasing recognition that individual risk from carcinogen exposure varies as a function of both inherited and acquired factors. Recent advances in genomics, microassay technologies and informatics hold promise for rapid identification of polymorphic variants or changes in expression of genes influencing both response and susceptibility to carcinogens. Another emerging area of molecular epidemiology concerns the role of nutrition and specific dietary factors (including studies on antioxidants, energy metabolism, insulin and various growth factors) and the modulating effect of genetic polymorphisms. Finally, molecular epidemiology has enormous potential in cancer prevention through the early identification of 'at risk' populations and the rapid assessment of intervention efficacy. Its success in fully reaching this potential will depend on the application of validated biomarkers, with adherence to sound epidemiologic and ethical principles. PMID- 10688873 TI - Chemoprevention of cancer. AB - In this short article, we review the conceptual basis for chemoprevention of cancer, the proven clinical efficacy of this concept, and current trends to develop new chemopreventive agents based on understanding of their mechanisms of action. Four classes of new agents, namely selective inhibitors of cyclooxygenase 2, selective estrogen receptor modulators, rexinoids (retinoids that bind selectively to the receptors known as RXRs) and ligands for the peroxisome proliferator-activated receptor-gamma are discussed in detail. The importance of developing totally new classes of chemopreventive agents is stressed, with particular emphasis on the potential usefulness of new synthetic triterpenoids derived from naturally occurring molecules. PMID- 10688874 TI - beta 3: an additional auxiliary subunit of the voltage-sensitive sodium channel that modulates channel gating with distinct kinetics. AB - The voltage-sensitive sodium channel confers electrical excitability on neurons, a fundamental property required for higher processes including cognition. The ion conducting alpha-subunit of the channel is regulated by two known auxiliary subunits, beta1 and beta2. We have identified rat and human forms of an additional subunit, beta3. It is most closely related to beta1 and is the product of a separate gene localized to human chromosome 11q23.3. When expressed in Xenopus oocytes, beta3 inactivates sodium channel opening more slowly than beta1 does. Structural modeling has identified an amino acid residue in the putative alpha-subunit binding site of beta3 that may play a role in this difference. The expression of beta3 within the central nervous system differs significantly from beta1. Our results strongly suggest that beta3 performs a distinct neurophysiological function. PMID- 10688875 TI - Spatial order within but not between types of retinal neurons. AB - We studied the mosaics of six types of retinal neurons, asking how the position of a cell relates to the positions of other cells of that same type and also to cells of different types. Every neuron studied was found to be nonrandomly positioned: Cells of a particular type were evenly spaced. However, all cells were positioned randomly with respect to members of the other cell classes. This was true even when the cells were known to be synaptically connected. It is consistent with a concept of developmental pattern formation in which (i) the number of cells of a particular type and their laminar distribution are specified, and (ii) the final spatial position of each cell is controlled exclusively by a rule that prevents cells of the same type from being positioned close to each other. This sequence would imply that a cell's final position is independent of the cell's position at the time of its specification, and we suggest a reason why, in laminar structures containing many cell types, it might be desirable for this to be so. PMID- 10688876 TI - Erectile dysfunction in cyclic GMP-dependent kinase I-deficient mice. AB - The generation of nitric oxide (NO) in penile erectile tissue and the subsequent elevation of cyclic GMP (cGMP) levels are important for normal penile erection. Current treatments of erectile dysfunction elevate either cGMP levels by blocking cGMP degrading phosphodiesterase 5 or cyclic AMP (cAMP) levels by intrapenile injection of prostaglandin E1. The molecular target or targets of cGMP in erectile tissue and the role of cAMP for normal penile erection are not known. Herein, we report that mice lacking cGMP-dependent kinase I (cGKI) have a very low ability to reproduce and that their corpora cavernosa fail to relax on activation of the NO/cGMP signaling cascade. Elevation of cAMP by forskolin, however, induces similar relaxation in normal and cGKI-null corpus cavernosum. In addition, sperm derived from cGKI-null mice is normal, can undergo acrosomal reactions, and can efficiently fertilize eggs. Altogether, these data identify cGKI as the downstream target of cGMP in erectile tissue and provide evidence that cAMP signaling cannot compensate for the absence of the cGMP/cGKI signaling cascade in vivo. PMID- 10688877 TI - Quantitative analysis of the effect of the mutation frequency on the affinity maturation of single chain Fv antibodies. AB - Random mutagenesis and selection using phage or cell surface display provides an efficient method for affinity maturation of single chain Fv (scFv) antibodies, thereby improving function in various applications. To investigate the effects of mutation frequency on affinity maturation, error-prone PCR was used to generate libraries containing an average (m) of between 1.7 and 22.5 base substitutions per gene in a high affinity scFv antibody that binds to the cardiac glycoside digoxigenin. The scFv antibody libraries were displayed on Escherichia coli, and mutant populations were analyzed by flow cytometry. At low to moderate mutation frequencies with an average mutation rate of m Ile) and beta282(Arg-->Gln)) located in the atomic model of alphabeta-tubulin near the taxane-binding site. Using molecular modeling, we investigated the conformational behavior of epothilone, which led to the identification of a common pharmacophore shared by taxanes and epothilones. Although two binding modes for the epothilones were predicted, one mode was identified as the preferred epothilone conformation as indicated by the activity of a potent pyridine-epothilone analogue. In addition, the structure-activity relationships of multiple taxanes and epothilones in the tubulin mutant cells can be fully explained by the model presented here, verifying its predictive value. Finally, these pharmacophore and activity data from mutant cells were used to model the tubulin binding of sarcodictyins, a distinct class of microtubule stabilizers, which in contrast to taxanes and the epothilones interact preferentially with the mutant tubulins. The unification of taxane, epothilone, and sarcodictyin chemistries in a single pharmacophore provides a framework to study drug-tubulin interactions that should assist in the rational design of agents targeting tubulin. PMID- 10688885 TI - Dynamic sensory sensitivity and children's word decoding skills. AB - The relationship between sensory sensitivity and reading performance was examined to test the hypothesis that the orthographic and phonological skills engaged in visual word recognition are constrained by the ability to detect dynamic visual and auditory events. A test battery using sensory psychophysics, psychometric tests, and measures of component literacy skills was administered to 32 unselected 10-year-old primary school children. The results suggest that children's sensitivity to both dynamic auditory and visual stimuli are related to their literacy skills. Importantly, after controlling for intelligence and overall reading ability, visual motion sensitivity explained independent variance in orthographic skill but not phonological ability, and auditory FM sensitivity covaried with phonological skill but not orthographic skill. These results support the hypothesis that sensitivity at detecting dynamic stimuli influences normal children's reading skills. Vision and audition separately may affect the ability to extract orthographic and phonological information during reading. PMID- 10688887 TI - Mammalian and chicken I forms of gonadotropin-releasing hormone in the gonads of a protochordate, Ciona intestinalis. AB - Two forms of gonadotropin-releasing hormone (GnRH) were isolated from the gonads of the tunicate, Ciona intestinalis. The primary structure of the purified peptides was determined by MS and chemical sequence analysis. Both GnRH forms have blocked NH(2) and COOH termini, and their primary structures are identical to mammalian (mGnRH) and chicken I (cGnRH-I) forms reported previously in vertebrates. A total of 1.2 mg of purified cGnRH-I and 0.98 mg of mGnRH was obtained from 100 g of Ciona gonads. The physiological effects of native GnRHs included the induction of synthesis and secretion of sex steroids from ciona gonads and the secretion of luteinizing hormone from rat pituitary. These results suggest that the primary structure and functional roles of mGnRH and cGnRH-I have been highly conserved throughout evolution of chordates. PMID- 10688886 TI - Tyrosine phosphorylation of p62dok by p210bcr-abl inhibits RasGAP activity. AB - The t(9;22) chromosomal translocation is found in almost all patients with chronic myelogenous leukemia. The resultant Bcr-Abl fusion gene expresses a chimeric fusion protein p210(bcr-abl) with increased tyrosine kinase activity. Hematopoietic progenitors isolated from chronic myelogenous leukemia patients in the chronic phase contain constitutively tyrosine-phosphorylated p62(dok) protein. p62(dok) associates with the Ras GTPase-activating protein (RasGAP), but only when p62(dok) is tyrosine phosphorylated. Here we have investigated the interaction between p62(dok) and RasGAP and the consequences of p62(dok) tyrosine phosphorylation on the activity of RasGAP. We have found that p62(dok) is directly tyrosine phosphorylated by p210(bcr-abl), and the sites of phosphorylation are located in the C-terminal half of the p62(dok) molecule. We have identified five tyrosine residues that are involved in in vitro RasGAP binding and have found that tyrosine-phosphorylated p62(dok) inhibits RasGAP activity. Our results suggest that p210(bcr-abl) might lead to the activation of the Ras signaling pathway by inhibiting a key down-regulator of Ras signaling. PMID- 10688889 TI - Light affects cAMP signaling and cell movement activity in Dictyostelium discoideum. AB - The multicellular, slug stage of the slime mould Dictyostelium discoideum lacks specific sensory cells and organs but can nevertheless respond in a very sensitive manner to external stimuli such as temperature and light. Within the migrating slug, the behavior of up to 100,000 individual amoebae is coordinated by cAMP mediated cell-cell signaling and chemotaxis. We report here the striking result that light directly modulates the cAMP cell-cell signaling system. Light induced secretion of cAMP from the slug tips decreased the period length of optical density waves and speeded up cell movement. A local effect of light on cAMP release within the slug tip could modulate cell movement within the slug and thus control its phototactic turning and orientation toward a light source. PMID- 10688888 TI - Molecular basis of a progressive juvenile-onset hereditary cataract. AB - In a recent paper, patients with a progressive juvenile-onset hereditary cataract have been reported to have a point mutation in the human gammaD crystallin gene (Stephan, D. A., Gillanders, E., Vanderveen, D., Freas-Lutz, D., Wistow, G., Baxevanis, A. D., Robbins, C. M., VanAuken, A., Quesenberry, M. I., Bailey Wilson, J., et al. (1999) Proc. Natl. Acad. Sci. USA 96, 1008-1012). This mutation results in the substitution of Arg-14 in the native protein by a Cys residue. It is not understood how this mutation leads to cataract. We have expressed recombinant wild-type human gammaD crystallin (HGD) and its Arg-14 to Cys mutant (R14C) in Escherichia coli and show that R14C forms disulfide-linked oligomers, which markedly raise the phase separation temperature of the protein solution. Eventually, R14C precipitates. In contrast, HGD slowly forms only disulfide-linked dimers and no oligomers. These data strongly suggest that the observed cataract is triggered by the thiol-mediated aggregation of R14C. The aggregation profiles of HGD and R14C are consistent with our homology modeling studies that reveal that R14C contains two exposed cysteine residues, whereas HGD has only one. Our CD, fluorescence, and differential scanning calorimetric studies show that HGD and R14C have nearly identical secondary and tertiary structures and stabilities. Thus, contrary to current views, unfolding or destabilization of the protein is not necessary for cataractogenesis. PMID- 10688890 TI - Carbonic anhydrase inhibitor suppresses invasion of renal cancer cells in vitro. AB - Acidification of the extracellular milieu of malignant tumors is reported to increase the invasive behavior of cancer cells. In normal tissues, production of acid is catalyzed by carbonic anhydrases (CAs), some of which are known to be overexpressed in certain cancers. To investigate the functional role of CA activity in such cancer cells, we analyzed the effect of acetazolamide, a potent CA inhibitor, on the invasive capacity of four renal carcinoma cell lines (Caki 1, Caki-2, ACHN, and A-498). We found that 10 microM acetazolamide inhibited the relative invasion rate of these cell lines between 18-74%. The Caki-2 and ACHN cell lines displayed the highest responsiveness, and their responses clearly depended on the acetazolamide concentration in the culture medium. Immunocytochemical and Western blotting results identified the presence of CA isoenzyme II in the cytoplasm of all four cell lines and CA XII on the plasma membrane in three of four cell lines. Because acetazolamide alone reduced invasiveness of these cancer cells in vitro, we conclude that the CAs overexpressed in these renal cancer cells contribute to invasiveness, at least in vitro, and suggest that CA inhibitors may also reduce invasiveness in other tumors that overexpress one or more CAs. PMID- 10688891 TI - Neuroimaging evidence implicating cerebellum in support of sensory/cognitive processes associated with thirst. AB - Recent studies implicate the cerebellum, long considered strictly a motor control structure, in cognitive, sensory, and affective phenomenon. The cerebellum, a phylogenetically ancient structure, has reciprocal ancient connections to the hypothalamus, a structure important in vegetative functions. The present study investigated whether the cerebellum was involved in vegetative functions and the primal emotions engendered by them. Using positron emission tomography, we examined the effects on the cerebellum of the rise of plasma sodium concentration and the emergence of thirst in 10 healthy adults. The correlation of regional cerebral blood flow with subjects' ratings of thirst showed major activation in the vermal central lobule. During the development of thirst, the anterior and posterior quadrangular lobule, lingula, and the vermis were activated. At maximum thirst and then during irrigation of the mouth with water to alleviate dryness, the cerebellum was less activated. However, 3 min after drinking to satiation, the anterior quadrangular lobule and posterior cerebellum were highly activated. The increased cerebellar activity was not related to motor behavior as this did not occur. Instead, responses in ancient cerebellar regions (vermis, fastigal nucleus, archicerebellum) may be more directly related to vegetative and affective aspects of thirst experiences, whereas activity in neocerebellar (posterior) regions may be related to sensory and cognitive aspects. Moreover, the cerebellum is apparently not involved in the computation of thirst per se but rather is activated during changes in thirst/satiation state when the brain is "vigilant" and is monitoring its sensory systems. Some neocerebellar activity may also reflect an intentionality for gratification by drinking inherent in the consciousness of thirst. PMID- 10688892 TI - Caspase-3: A vulnerability factor and final effector in apoptotic death of dopaminergic neurons in Parkinson's disease. AB - Caspase-3 is an effector of apoptosis in experimental models of Parkinson's disease (PD). However, its potential role in the human pathology remains to be demonstrated. Using caspase-3 immunohistochemistry on the postmortem human brain, we observed a positive correlation between the degree of neuronal loss in dopaminergic (DA) cell groups affected in the mesencephalon of PD patients and the percentage of caspase-3-positive neurons in these cell groups in control subjects and a significant decrease of caspase-3-positive pigmented neurons in the substantia nigra pars compacta of PD patients compared with controls that also could be observed in an animal model of PD. This suggests that neurons expressing caspase-3 are more sensitive to the pathological process than those that do not express the protein. In addition, using an antibody raised against activated caspase-3, the percentage of active caspase-3-positive neurons among DA neurons was significantly higher in PD patients than in controls. Finally, electron microscopy analysis in the human brain and in vitro data suggest that caspase-3 activation precedes and is not a consequence of apoptotic cell death in PD. PMID- 10688893 TI - Membrane hyperpolarization removes inactivation of Ca2+ channels, leading to Ca2+ influx and subsequent initiation of sperm motility in the common carp. AB - Change of osmolality surrounding spawned sperm from isotonic to hypotonic causes the initiation of sperm motility in the common carp. Here we show that membrane permeable cAMP does not initiate motility of carp sperm that is quiescent in isotonic solution, and that motility of the demembranated sperm can be reactivated without cAMP. Furthermore, the cAMP level does not change during the initiation of sperm motility, and inhibitors of protein kinase do not affect sperm motility, suggesting that no cAMP-dependent system is necessary for the regulation of sperm motility. Sperm motility could not be initiated in Ca(2+) free hypoosmotic solutions, and significant increase in the intracellular Ca(2+) level was observed by a Ca-sensitive fluorescence dye during hypoosmolality induced active motion period. The demembranated sperm cells were fully reactivated in the solutions containing 10(-7) to 10(-5) M Ca(2+). Ca(2+) channel blockers such as verapamil and omega-conotoxin reversibly inhibited the initiation of sperm motility, suggesting that Ca(2+) influx is the prerequisite for the initiation of carp sperm motility. Motility of intact sperm was completely blocked; however, that of the demembranated sperm was not inhibited by the calmodulin inhibitor W7, suggesting that the calmodulin bound close to the plasma membrane participated in the initiation of sperm motility. Flow cytometric membrane potential measurements and spectrophotometric measurements by using fluorescence dyes showed transient membrane hyperpolarization on hypoosmolality induced motility. This article discusses the role of membrane hyperpolarization on removal of inactivation of Ca(2+) channels, leading to Ca(2+) influx at the initiation of carp sperm motility. PMID- 10688894 TI - Viral persistence in vivo through selection of neutralizing antibody-escape variants. AB - Despite initial virus control by CD8(+) cytotoxic T lymphocytes (CTLs), noncytopathic or variably cytopathic viruses (e.g., hepatitis B and C viruses, HIV) are able to establish persistent infections. The role of neutralizing antibodies (nAbs) in controlling disease progression is unclear. Therefore, the phenomenon of viral evasion from the nAb response and its implications for virus persistence remain controversial. Here we demonstrate nAb-mediated viral clearance in CTL-deficient mice infected with the prototypic noncytopathic lymphocytic choriomeningitis virus (strain WE). During prolonged CTL absence, neutralization-resistant virus mutants were selected in individual mice within 70 90 days. In naive animals infected with these virus variants only low nAb responses were induced, resulting in an increased tendency of virus to persist. PMID- 10688895 TI - Marking synaptic activity in dendritic spines with a calpain substrate exhibiting fluorescence resonance energy transfer. AB - Excitatory synaptic activity can evoke transient and substantial elevations of postsynaptic calcium. Downstream effects of elevated calcium include the activation of the calcium-dependent protease calpain. We have developed a reagent that identifies dendritic spines in which calpain has been activated. A fusion protein was expressed that contained enhanced yellow and enhanced cyan fluorescent protein (EYFP and ECFP, respectively) linked by a peptide that included the micro-calpain cleavage site from alpha-spectrin. A PDZ-binding site fused to ECFP anchored this protein to postsynaptic densities. The fusion protein exhibited fluorescence resonance energy transfer (FRET), and diminution of FRET by proteolysis was used to localize calpain activity in situ by fluorescence microscopy. Incubation of the fusion protein with calpain in the presence of calcium resulted in the separation of EYFP and ECFP into monomeric fluorophores. In transiently transfected cell lines and dissociated hippocampal neurons, FRET was diminished by raising intracellular calcium levels with an ionophore or with glutamatergic agonists. Calpain inhibitors blocked these changes. Under control conditions, FRET levels in different dendritic spines of cultured neurons and in hippocampal slices were heterogeneous but showed robust decreases upon treatment with glutamatergic agonists. Immunostaining of cultured neurons with antibodies to a spectrin epitope produced by calpain-mediated digestion revealed an inverse correlation between the amount of FRET present at postsynaptic elements and the concentration of spectrin breakdown products. These results suggest that the FRET methodology identifies sites of synaptically induced calpain activity and that it may be useful in analyzing synapses undergoing changes in efficacy. PMID- 10688896 TI - Inhibitory pathways and the inhibition of luteinizing hormone-releasing hormone release by alcohol. AB - In this research we examined the mechanisms by which ethanol (EtOH) inhibits luteinizing hormone-releasing hormone (LHRH) release from incubated medial basal hypothalamic explants. EtOH (100 mM) stimulated the release of two inhibitory neurotransmitters: gamma-aminobutyric acid (GABA) and beta-endorphin. EtOH also inhibited NO production, indicative of a suppression of nitric oxide synthase (NOS) activity. This inhibition was reversed by naltroxone (10(-8) M), a micro opioid receptor blocker, indicating that the inhibition of NOS by EtOH is mediated by beta-endorphin. EtOH also blocked N-methyl-d-aspartic acid-induced LHRH release, but the blockade could not be reversed by either the GABA receptor blocker, bicuculline (10(-5) M), naltroxone (10(-8) M), or both inhibitors added together. However, increasing the concentration of naltrexone (10(-6) M) but not bicuculline (10(-4) M) reversed the inhibition. When we lowered the concentration of EtOH (50 mM), the EtOH-induced blockade of LHRH release could be reversed by either bicuculline (10(-5) M), naltroxone (10(-8) M), or the combination of the two blockers. Therefore, GABA is partially responsible for the blockade of N methyl-d-aspartic acid-induced LHRH release. The block by GABA was exerted by inhibiting the activation of cyclooxygenase by NO, because it was reversed by prostaglandin E(2), the product of activation of cyclooxygenase. Because the inhibition caused by the higher concentration of EtOH could not be reduced by bicuculline (10(-4) M) but was blocked by naltroxone (10(-6) M), the action of alcohol can be accounted for by stimulation of beta-endorphin neurons that inhibit LHRH release by inhibition of activation of NOS and stimulation of GABA release. PMID- 10688897 TI - The structure of a stable intermediate in the A <--> B DNA helix transition. AB - The DNA dodecamer CATGGGCCCATG in a crystal structure of resolution 1.3 A has a conformation intermediate between A and B DNA. This trapping of a stable intermediate suggests that the A and B DNA families are not discrete, as previously believed. The structure supports a base-centered rather than a backbone-centered mechanism for the A <--> B transition mediated by guanine tracts. Interconversion between A and B DNA provides another means for regulating protein-DNA recognition. PMID- 10688899 TI - Isolation of peptide aptamers that inhibit intracellular processes. AB - We have developed a method for isolation of random peptides that inhibit intracellular processes in bacteria. A library of random peptides expressed as fusions to Escherichia coli thioredoxin (aptamers) were expressed under the tight control of the arabinose-inducible P(BAD) promoter. A selection was applied to the library to isolate aptamers that interfered with the activity of thymidylate synthase (ThyA) in vivo. Expression of an aptamer isolated by this method resulted in a ThyA(-) phenotype that was suppressed by simultaneous overexpression of ThyA. Two-hybrid analysis showed that this aptamer is likely to interact with ThyA in vivo. The library also was screened for aptamers that inhibited growth of bacteria expressing them, and five such aptamers were characterized. Four aptamers were bacteriostatic when expressed, whereas one showed a bactericidal effect. Introduction of translational stop codons into various aptamers blocked their activity, suggesting that their biological effects were likely to be due to protein aptamer rather than RNA. Combinatorial aptamers provide a new genetic and biochemical tool for identifying targets for antibacterial drug development. PMID- 10688898 TI - The EntF and EntE adenylation domains of Escherichia coli enterobactin synthetase: sequestration and selectivity in acyl-AMP transfers to thiolation domain cosubstrates. AB - Enterobactin, the tris-(N-(2,3-dihydroxybenzoyl)serine) trilactone siderophore of Escherichia coli, is synthesized by a three-protein (EntE, B, F) six-module nonribosomal peptide synthetase (NRPS). In this work, the 142-kDa four-domain protein EntF was bisected into two double-domain fragments: a 108-kDa condensation and adenylation construct, EntF C-A, and a 37-kDa peptidyl carrier protein (PCP) and thioesterase protein, EntF PCP-TE. The adenylation domain activity of EntF C-A formed seryl-AMP but lost the ability to transfer the seryl moiety to the cognate EntF PCP-TE in trans. Seryl transfer to heterologous PCP protein fragments, the SrfB1 PCP from surfactin synthetase and Ybt PCP1 from yersiniabactin synthetase, was observed at rates of 0.5 min(-1) and 0.01 min(-1), respectively. The possibility that these slow acylation rates reflected dissociation of acyl/aminoacyl-AMP followed by adventitious thiolation by the heterologous PCPs in solution was addressed by measuring catalytic turnover of pyrophosphate (PP(i)) released from the adenylation domain. The holo SrfB1 PCP protein as well as Ybt PCP1 did not stimulate an increase in PP(i) release from EntF C-A or EntE. In this light, aminoacylations in trans between A and PCP domain fragments of NRPS assembly lines must be subjected to kinetic scrutiny to determine whether transfer is truly between protein domains or results from slow aminoacyl-AMP release and subsequent nonenzymatic thiol capture. PMID- 10688900 TI - The amino terminus of the mixed lineage leukemia protein (MLL) promotes cell cycle arrest and monocytic differentiation. AB - Several lines of evidence suggest that the mixed lineage leukemia protein (MLL, ALL-1, HRX) plays a role in regulating myelomonocytic differentiation. In this study we examined the effect of expression of MLL-AF9 on differentiation of the monoblastic U937 cell line by using a tetracycline-inducible expression system. MLL-AF9 arrested growth of U937 cells and induced these cells to differentiate into macrophages; induction was accompanied by expression of CD11b and CD14 and ultimately cell death. Deletion mutants of MLL-AF9 were used to map the sequences responsible for this effect. The amino-terminal half of MLL was sufficient for both cell cycle arrest and macrophage differentiation, whereas the carboxyl terminus of MLL or AF9 was found to be dispensable for this effect. Further deletions showed that a 35-kDa amino-terminal fragment spanning two AT hook motifs was sufficient for cell cycle arrest, up-regulation of p21(Cip1) and p27(Kip1), and partial differentiation toward macrophages. These findings suggest a possible role for the MLL AT hook-containing region in regulating myelomonocytic differentiation. PMID- 10688901 TI - The B cell-restricted adaptor BASH is required for normal development and antigen receptor-mediated activation of B cells. AB - B cell antigen receptor signals development, activation, proliferation, or apoptosis of B cells depending on their condition, and its proper signaling is critical for activation and homeostasis of the immune system. The B cell restricted adaptor protein BASH (also termed BLNK/SLP-65) is rapidly phosphorylated by the tyrosine kinase Syk after BCR ligation and binds to various signaling proteins. BASH structurally resembles SLP-76, which is essential for T cell development and T cell receptor signaling. To evaluate the role for BASH in B cell development and function in vivo, we disrupted BASH alleles in embryonic stem cells by means of homologous recombination and used these cells to complement lymphocyte-incompetent blastocysts from RAG2-deficient mice. In the resultant chimeric mice, T cell development was apparently normal, but B cell development was impaired, and a normally rare population of large preB cells expressing preB cell receptor dominated in the bone marrow in place of small preB cells, although they were mostly noncycling. In addition, the mature B cell populations in the periphery and the bone marrow profoundly decreased in size, as did B-1 cells in the peritoneal cavity, and serum Ig was severely reduced. The BASH-deficient B cells scarcely proliferated or up-regulated B7-2 in response to BCR ligation and poorly proliferated upon CD40 ligation or lipopolysaccharide stimulation. This phenotype indicates that BASH is critical for preB cell receptor signaling inducing proliferation of large preB cells and the following differentiation, for peripheral B cell maturation, and for BCR signaling inducing activation/proliferation of B cells. PMID- 10688902 TI - Place recognition monitored by location-driven operant responding during passive transport of the rat over a circular trajectory. AB - Spatial memory of animals is usually tested in navigation tasks that do not allow recognition and recall processes to be separated from the mechanisms of goal directed locomotion. In the present study, place recognition was examined in rats (n = 7) confined in an operant chamber mounted on the periphery of a slowly rotating disk (diameter 1 m, angular velocity 9 degrees /s). The animals were passively transported over a circular trajectory and were rewarded for bar pressing when they passed across a 60 degrees -wide segment of the path. This segment was recognizable with reference to room landmarks visible from the operant box. Responding defined in the coordinate system of the room increased when the chamber entered the 60 degrees -wide approach zone, culminated at the entrance into the reward sector, was decreased inside it by eating the available reward, and rapidly declined to zero at the exit from this zone. When reward was discontinued, the skewed response distribution changed into a symmetric one with a maximum in the center of the reward sector. With advancing extinction, the response peak in the reward sector decreased in most rats proportionally to the overall decline of bar pressing. The rewarded and nonrewarded response patterns indicate that passively transported rats can recognize their position in the environment with an accuracy comparable to that of actively navigating animals and that location-driven operant responding can serve as a useful tool in the analysis of the underlying neural mechanisms. PMID- 10688904 TI - Candidate tumor suppressor RIZ is frequently involved in colorectal carcinogenesis. AB - The distal portion of chromosome 1p is one of the most commonly affected regions in human cancer. In this study of hereditary and sporadic colorectal cancer, a region of frequent deletion was identified at 32.2 centimorgans from 1ptel. Deletion breakpoints clustered in the vicinity of or inside the gene RIZ, which encodes a retinoblastoma protein-interacting zinc finger protein. Sequence analysis revealed frequent frameshift mutations of the RIZ gene. The mutations consisted of 1- or 2-bp deletions of a coding (A)(8) or (A)(9) tract and were confined to microsatellite-unstable colorectal tumors, being present in 9 of 24 (37.5%) primary tumors and in 6 of 11 (54.5%) cell lines; in 2 cell lines the mutation was homozygous/hemizygous. The mutations apparently were selected clonally in tumorigenesis, because similar poly(A) tracts in other genes were not affected. Two alternative products of the gene exist, RIZ1, which contains a PR (PRDI-BF1-RIZ1) domain implicated in tumor suppressor function, and RIZ2, which is lacking this motif. Furthermore, the C-terminal region, which contains the poly(A) tracts, includes a PR-binding motif, possibly mediating interactions with other proteins or with RIZ itself (oligomerization). Four of eleven microsatellite-unstable colorectal cancer cell lines, three of which had frameshifts, showed reduced or absent mRNA expression of RIZ1. In a cell line that is homozygous/hemizygous for the typical frameshift mutation, immunoblotting showed truncated RIZ protein, whereas adenovirus-mediated RIZ1 expression caused G(2)/M arrest and apoptosis. We propose that RIZ is a target of the observed 1p alterations, with impairment of the PR domain-mediated function through either frameshift mutation or genomic deletion. PMID- 10688905 TI - 5-hydroxytryptamine 2B receptor regulates cell-cycle progression: cross-talk with tyrosine kinase pathways. AB - In this paper, we present evidence that activation of 5-hydroxytryptamine 2B (5 HT2B) receptors by serotonin (5-HT) leads to cell-cycle progression through retinoblastoma protein hyperphosphorylation and through activation of both cyclin D1/cdk4 and cyclin E/cdk2 kinases by a mechanism that depends on induction of cyclin D1 and cyclin E protein levels. The induction of cyclin D1 expression, but not that of cyclin E, is under mitogen-activated protein kinase (MAPK) control, indicating an independent regulation of these two cyclins in the 5-HT2B receptor mitogenesis. Moreover, by using the specific platelet-derived growth factor receptor (PDGFR) inhibitor AG 1296 or by overexpressing a kinase-mutant PDGFR, we show that PDGFR kinase activity is essential for 5-HT2B-triggered MAPK/cyclin D1, but not cyclin E, signaling pathways. 5-HT2B receptor activation also increases activity of the Src family kinase, c-Src, Fyn, and c-Yes. Strikingly, c-Src, but not Fyn or c-Yes, is the crucial molecule between the G(q) protein-coupled 5-HT2B receptor and the cell-cycle regulators. Inhibition of c-Src activity by 4-amino-5 (4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP1) or depletion of c-Src is sufficient to abolish the 5-HT-induced (i) PDGFR tyrosine kinase phosphorylation and MAPK activation, (ii) cyclin D1 and cyclin E expression levels, and (iii) thymidine incorporation. This paper elucidates a model of 5 HT2B receptor mitogenesis in which c-Src acts alone to control cyclin E induction and in concert with the receptor tyrosine kinase PDGFR to induce cyclin D1 expression via the MAPK/ERK pathway. PMID- 10688906 TI - A B-cell receptor-specific selection step governs immature to mature B cell differentiation. AB - Seventy percent of peripheral immature conventional (B2) B cells fail to develop into mature B cells. The nature of this cell loss has not been characterized; the process that governs which immature B cells develop into long-lived peripheral B cells could be either stochastic or selective. Here, we demonstrate that this step is in fact selective, in that the fate of an immature B cell is highly dependent on its Ig receptor specificity. A significant skewing of the B cell receptor repertoire occurs by the time cells enter the mature B cell fraction, which indicates that there is selection of only a minority of immature B cells to become mature B cells. Because only a few heavy-light chain pairs are enhanced of the diverse available repertoire, we favor the idea that selection is positive for these few heavy-light chain pairs rather than negative against nearly all others. Because most immature B cells are lost at this transition, this putative positive selection event is likely to be a major force shaping the mature B cell receptor repertoire available for adaptive immune responses. PMID- 10688907 TI - Rat strain-specific actions of 17beta-estradiol in the mammary gland: correlation between estrogen-induced lobuloalveolar hyperplasia and susceptibility to estrogen-induced mammary cancers. AB - The genetically related ACI and Copenhagen (COP) rat strains display diametrically opposed susceptibilities to mammary cancer development when treated chronically with 17beta-estradiol (E2). Here, we compare the actions of E2 on cell proliferation and lobuloalveolar development in the mammary glands of female ACI and COP rats. After 12 wk of E2 treatment, the mammary glands of ACI rats exhibited a significantly greater proliferative response to E2, compared with COP rats, as evidenced by quantification of S phase fraction and development of lobuloalveolar hyperplasia. Focal regions of atypical epithelial hyperplasia were observed in ACI, but not COP, rats. These strain differences were not because of differences in circulating E2, progesterone or, prolactin. Two-thirds of the induced mammary cancers in ACI rats exhibited aneuploidy. The E2-induced mammary cancers regressed when hormone treatment was discontinued, indicating that they were estrogen-dependent. Progesterone receptor was expressed by the great majority of epithelial cells within the E2-induced atypical hyperplastic foci and the mammary carcinomas, suggesting a link between these lesions. These data demonstrate a correlation between E2 action in the induction of mammary cell proliferation and atypical epithelial hyperplasia and genetically conferred susceptibility to E2-induced mammary cancers. PMID- 10688908 TI - Expression and function of wingless and frizzled homologs in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is accompanied by synovial inflammation, proliferation, and cartilage destruction. The reasons the activation of synovial fibroblasts often persists despite antiinflammatory therapy are not known. One possibility is that the synovial membrane becomes gradually repopulated with immature mesenchymal and bone marrow cells with altered properties. To explore this hypothesis, we have investigated the expression in RA synovial tissues of various embryonic growth factors from the wingless (wnt) and frizzled (fz) families, which have been implicated in cell-fate determination in both bone marrow progenitors and limb-bud mesenchyme. Reverse transcriptase-PCR analysis revealed expression of five wnt (wnt1, 5a, 10b, 11, and 13) and three fz (fz2, 5, and 7) isoforms in RA synovial tissues. Osteoarthritis synovial tissues expressed much less wnt5a and fz5. Northern blotting confirmed the overexpression of wnt5a and fz5 in RA synovial tissues, in comparison to a panel of normal adult tissues. Compared with normal synovial fibroblasts, cultured RA fibroblast-like synoviocytes expressed higher levels of IL-6, IL-8, and IL-15. Transfection of normal fibroblasts with a wnt5a expression vector reproduced this pattern of cytokine expression and stimulated IL-15 secretion. These results suggest that the unusual phenotypic properties of RA fibroblasts may be attributable partly to their replacement with primitive fibroblast-like synoviocytes with characteristics of immature bone marrow and mesenchymal cells. Clear delineation of the signaling pathway(s) initiated by the wnt5a/fz5 ligand-receptor pair in the RA synovium may yield new targets for therapeutic intervention. PMID- 10688909 TI - The short interspersed repetitive element of Trypanosoma cruzi, SIRE, is part of VIPER, an unusual retroelement related to long terminal repeat retrotransposons. AB - The short interspersed repetitive element (SIRE) of Trypanosoma cruzi was first detected when comparing the sequences of loci that encode the TcP2beta genes. It is present in about 1,500-3,000 copies per genome, depending on the strain, and it is distributed in all chromosomes. An initial analysis of SIRE sequences from 21 genomic fragments allowed us to derive a consensus nucleotide sequence and structure for the element, consisting of three regions (I, II, and III) each harboring distinctive features. Analysis of 158 transcribed SIREs demonstrates that the consensus is highly conserved. The sequences of 51 cDNAs show that SIRE is included in the 3' end of several mRNAs, always transcribed from the sense strand, contributing the polyadenylation site in 63% of the cases. This study led to the characterization of VIPER (vestigial interposed retroelement), a 2,326-bp long unusual retroelement. VIPER's 5' end is formed by the first 182 bp of SIRE, whereas its 3' end is formed by the last 220 bp of the element. Both SIRE moieties are connected by a 1,924-bp-long fragment that carries a unique ORF encoding a complete reverse transcriptase-RNase H gene whose 15 C-terminal amino acids derive from codons specified by SIRE's region II. The amino acid sequence of VIPER's reverse transcriptase-RNase H shares significant homology to that of long terminal repeat retrotransposons. The fact that SIRE and VIPER sequences are found only in the T. cruzi genome may be of relevance for studies concerning the evolution and the genome flexibility of this protozoan parasite. PMID- 10688910 TI - Differential regulation by multiple promoters of the gene encoding the neuron restrictive silencer factor. AB - NRSF/REST is a protein that silences transcription of a number of genes that contain a DNA element called the neuron-restrictive silencer element (NRSE). During embryogenesis, REST is expressed ubiquitously in nonneural cells, but is down-regulated during differentiation of neural progenitors into neurons. REST is also up-regulated in adult neurons by activity, suggesting a possible role for the protein in synaptic plasticity. To understand mechanisms that control expression of REST, we identified and characterized the promoter region of the mouse REST gene (mREST). A 4.5-kb DNA segment containing three exons (A, B, and C) that correspond to alternatively spliced 5' untranslated regions (5'UTRs) was isolated and its DNA sequence was determined. Reverse transcription-PCR analyses of fibroblasts, astrocytes, and neural progenitors identified variants in which these exons were spliced to exon D, suggesting that exons A, B, and C may each have a promoter. Consistent with this hypothesis, primer extension and in vitro transcription experiments revealed clusters of RNA transcription initiation sites upstream of exons A, B, and C. Tests of REST/luciferase reporter constructs in Neuro2A and NIH 3T3 cells revealed promoters upstream of exons A and B that were active in both cell lines, and a promoter upstream of exon C that was weakly active only in NIH 3T3 cells. Six enhancer and two repressor regions were found to overlap each of the three promoters, and some of these were found to be cell type-specific. Combinatorial arrangements of these promoters with enhancer and repressor regions may allow modulation of REST expression in particular contexts. PMID- 10688911 TI - Mammalian thioredoxin reductase: oxidation of the C-terminal cysteine/selenocysteine active site forms a thioselenide, and replacement of selenium with sulfur markedly reduces catalytic activity. AB - Mammalian cytosolic thioredoxin reductase (TrxR) has a redox center, consisting of Cys(59)/Cys(64) adjacent to the flavin ring of FAD and another center consisting of Cys(497)/selenocysteine (SeCys)(498) near the C terminus. We now show that the C-terminal Cys(497)-SH/SeCys(498)-Se(-) of NADPH-reduced enzyme, after anaerobic dialysis, was converted to a thioselenide on incubation with excess oxidized Trx (TrxS(2)) or H(2)O(2). The Cys(59)-SH/Cys(64)-SH pair also was oxidized to a disulfide. At lower concentrations of TrxS(2), the Cys(59) SH/Cys(64)-SH center was still converted to a disulfide, presumably by reduction of the thioselenide to Cys(497)-SH/SeCys(498)-Se(-). Specific alkylation of SeCys(498) completely blocked the TrxS(2)-induced oxidation of Cys(59)-SH/Cys(64) SH, and the alkylated enzyme had negligible NADPH-disulfide oxidoreductase activity. The effect of replacing SeCys(498) with Cys was determined by using a mutant form of human placental TrxR1 expressed in Escherichia coli. The NADPH disulfide oxidoreductase activity of the purified Cys(497)/Cys(498) mutant enzyme was 6% or 11% of that of wild-type rat liver TrxR1 with 5, 5'-dithiobis(2 nitrobenzoic acid) or TrxS(2), respectively, as substrate. Disulfide formation induced by excess TrxS(2) in the mutant form was 12% of that of the wild type. Thus, SeCys has a critical redox function during the catalytic cycle, which is performed poorly by Cys. PMID- 10688912 TI - Selective interaction between leptin and insulin signaling pathways in a hepatic cell line. AB - Leptin is a 16-kDa hormone secreted by adipocytes and plays an important role in control of feeding behavior and energy expenditure. In obesity, circulating levels of leptin and insulin are high because of the presence of increased body fat mass and insulin resistance. Recent reports have suggested that leptin can act through some of the components of the insulin signaling cascade, such as insulin receptor substrates (IRS-1 and IRS-2), phosphatidylinositol 3-kinase (PI 3-kinase), and mitogen-activated protein kinase, and can modify insulin-induced changes in gene expression in vitro and in vivo. Well differentiated hepatoma cells (Fao) possess both the long and short forms of the leptin receptor and respond to leptin with a stimulation of c-fos gene expression. In Fao cells, leptin alone had no effects on the insulin signaling pathway, but leptin pretreatment transiently enhanced insulin-induced tyrosine phosphorylation and PI 3-kinase binding to IRS-1, while producing an inhibition of tyrosine phosphorylation and PI 3-kinase binding to IRS-2. Leptin alone also induced serine phosphorylation of Akt and glycogen synthase kinase 3 but to a lesser extent than insulin, and the combination of these hormones was not additive. These results suggest complex interactions between the leptin and insulin signaling pathways that can potentially lead to differential modification of the metabolic and mitotic effects of insulin exerted through IRS-1 and IRS-2 and the downstream kinases that they activate. PMID- 10688913 TI - Recombinant adeno-associated virus type 2, 4, and 5 vectors: transduction of variant cell types and regions in the mammalian central nervous system. AB - Recombinant adeno-associated virus vectors based on serotype 2 (rAAV2) can direct transgene expression in the central nervous system (CNS), but it is not known how other rAAV serotypes perform as CNS gene transfer vectors. Serotypes 4 and 5 are distinct from rAAV2 and from each other in their capsid regions, suggesting that they may direct binding and entry into different cell types. In this study, we examined the tropisms and transduction efficiencies of beta-galactosidase encoding vectors made from rAAV4 and rAAV5 compared with similarly designed rAAV2 based vectors. Injection of rAAV5 beta-galactosidase (betagal) or rAAV4betagal into the lateral ventricle resulted in stable transduction of ependymal cells, with approximately 10-fold more positive cells than in mice injected with rAAV2betagal. Major differences between the three vectors were revealed upon striatal injections. Intrastriatal injection of rAAV4betagal resulted again in striking ependyma-specific expression of transgene, with a notable absence of transduced cells in the parenchyma. rAAV2betagal and rAAV5betagal intrastriatal injections led to beta-gal-positive parenchymal cells, but, unlike rAAV2betagal, rAAV5betagal transduced both neurons and astrocytes. The number of transgene positive cells in rAAV5betagal-injected brains was 130 and 5,000 times higher than in rAAV2betagal-injected brains at 3 and 15 wk, respectively. Moreover, transgene-positive cells were widely dispersed throughout the injected hemisphere in rAAV5betagal-transduced animals. Together, our data provide in vivo support for earlier in vitro work, suggesting that rAAV4 and rAAV5 gain cell entry by means of receptors distinct from rAAV2. These differences could be exploited to improve gene therapy for CNS disorders. PMID- 10688914 TI - Posttranscriptional regulation of Bruton's tyrosine kinase expression in antigen receptor-stimulated splenic B cells. AB - Mutation of Bruton's tyrosine kinase (Btk) causes human X-linked agammaglobulinemia and murine X-linked immunodeficiency syndrome (xid). Quantitative aspects of B lymphocyte development and function have been demonstrated to depend on Btk level in vivo by using a murine transgenic model system. A sensitive intracellular immunofluorescent assay was developed to measure Btk protein on a per cell basis to test the hypothesis that its dosage is dynamically regulated during B cell development or functional responses. Marrow derived hematopoietic stem cells, common lymphoid progenitor cells, and developing B and myeloid lineages expressed Btk protein at comparable levels. Resting peripheral B lineage cells had a significantly lower amount of Btk than marrow-derived cells in both wild-type and xid mice. Activation of the B cell antigen receptor up-regulated Btk protein level 10-fold within several hours by a phosphatidylinositol 3-kinase-dependent, posttranscriptional mechanism. In contrast, the protein level of Btk R28C in activated B lymphocytes from xid mice remained low. Bypass of the antigen receptor signaling pathways by treatment of cells with phorbol myristic acid and ionomycin rescued up-regulation of Btk protein in xid splenic B cells. These combined results suggest that certain receptor signals mediated by Btk regulate the level of expression of Btk protein in responding B lymphocytes to potentiate signal transduction. Dynamic regulation of Btk protein dosage is an additional mechanism to modulate B lymphocyte immune functions. PMID- 10688915 TI - Identification of CDK4 as a target of c-MYC. AB - The prototypic oncogene c-MYC encodes a transcription factor that can drive proliferation by promoting cell-cycle reentry. However, the mechanisms through which c-MYC achieves these effects have been unclear. Using serial analysis of gene expression, we have identified the cyclin-dependent kinase 4 (CDK4) gene as a transcriptional target of c-MYC. c-MYC induced a rapid increase in CDK4 mRNA levels through four highly conserved c-MYC binding sites within the CDK4 promoter. Cell-cycle progression is delayed in c-MYC-deficient RAT1 cells, and this delay was associated with a defect in CDK4 induction. Ectopic expression of CDK4 in these cells partially alleviated the growth defect. Thus, CDK4 provides a direct link between the oncogenic effects of c-MYC and cell-cycle regulation. PMID- 10688916 TI - Chip interacts with diverse homeodomain proteins and potentiates bicoid activity in vivo. AB - The Drosophila protein Chip potentiates activation by several enhancers and is required for embryonic segmentation. Chip and its mammalian homologs interact with and promote dimerization of nuclear LIM proteins. No known Drosophila LIM proteins, however, are required for segmentation, nor for expression of most genes known to be regulated by Chip. Here we show that Chip also interacts with diverse homeodomain proteins using residues distinct from those that interact with LIM proteins, and that Chip potentiates activity of one of these homeodomain proteins in Drosophila embryos and in yeast. These and other observations help explain the roles of Chip in segmentation and suggest a model to explain how Chip potentiates activation by diverse enhancers. PMID- 10688917 TI - Activation of antitumor cytotoxic T lymphocytes by fusions of human dendritic cells and breast carcinoma cells. AB - We have reported that fusions of murine dendritic cells (DCs) and murine carcinoma cells reverse unresponsiveness to tumor-associated antigens and induce the rejection of established metastases. In the present study, fusions were generated with primary human breast carcinoma cells and autologous DCs. Fusion cells coexpressed tumor-associated antigens and DC-derived costimulatory molecules. The fusion cells also retained the functional potency of DCs and stimulated autologous T cell proliferation. Significantly, the results show that autologous T cells are primed by the fusion cells to induce MHC class I-dependent lysis of autologous breast tumor cells. These findings demonstrate that fusions of human breast cancer cells and DCs activate T cell responses against autologous tumors. PMID- 10688919 TI - Serotonergic control of developmental plasticity. PMID- 10688920 TI - Building a comprehensive, evidence based tobacco treatment system in managed care PMID- 10688918 TI - Physical association of ubiquitin ligases and the 26S proteasome. AB - The ubiquitin (Ub) system recognizes degradation signals of the target proteins through the E3 components of E3-E2 Ub ligases. A targeted substrate bears a covalently linked multi-Ub chain and is degraded by the ATP-dependent 26S proteasome, which consists of the 20S core protease and two 19S particles. The latter mediate the binding and unfolding of a substrate protein before its transfer to the interior of the 20S core. It is unclear how a targeted substrate is delivered to the 26S proteasome, inasmuch as Rpn10p, the only known proteasomal subunit that binds multi-Ub chains, has been found to be not essential for degradation of many proteins in the yeast Saccharomyces cerevisiae. Here we show that Ubr1p and Ufd4p, the E3 components of two distinct Ub ligases, directly interact with the 26S proteasome. Specifically, Ubr1p is shown to bind to the Rpn2p, Rpt1p, and Rpt6p proteins of the 19S particle, and Ufd4p is shown to bind to Rpt6p. These and related results suggest that a substrate-bound Ub ligase participates in the delivery of substrates to the proteasome, because of affinity between the ligase's E3 component and specific proteins of the 19S particle. PMID- 10688921 TI - Addressing tobacco in managed care conference PMID- 10688922 TI - Tobacco dependence treatment: scientific challenges; public health opportunities. PMID- 10688923 TI - Best practices for comprehensive tobacco control programs: opportunities for managed care organisations. PMID- 10688924 TI - A proactive health plan: taking action on tobacco control. PMID- 10688925 TI - Managed care and the state tobacco settlements. PMID- 10688926 TI - Implementing tobacco interventions in the real world of managed care. PMID- 10688927 TI - Designing tobacco control systems and cessation benefits in managed care: skill building workshop. PMID- 10688928 TI - Tobacco cessation program implementation-from plans to reality: skill building workshop-group model. PMID- 10688929 TI - Tobacco cessation program implementation-from plans to reality: skill building workshop-network model. PMID- 10688930 TI - Incentivising, facilitating, and implementing an office tobacco cessation system. PMID- 10688931 TI - Implementing tobacco tracking codes in an individual practice association or a network model health maintenance organisation. PMID- 10688932 TI - How a real time clinical data retrieval system might be applied to a tobacco cessation program. PMID- 10688933 TI - Findings from the Addressing Tobacco in Managed Care focus groups: an executive summary. PMID- 10688934 TI - I. Tailored communications for smoking cessation. Introduction. PMID- 10688935 TI - Facilitating smoking cessation in cancer patients. PMID- 10688936 TI - Tailoring tobacco control messages for Hispanic populations. PMID- 10688937 TI - Communicating with teens: some lessons from commercial marketing. PMID- 10688938 TI - Computer tailored intervention for older smokers using transdermal nicotine. PMID- 10688939 TI - II. Smoking cessation in the hospital setting-a new opportunity for managed care. Introduction. PMID- 10688940 TI - Hospitalised smokers: characteristics, treatment, and transition to ambulatory care. PMID- 10688941 TI - Smoking cessation interventions for patients with heart disease. PMID- 10688942 TI - III. Maternal smoking cessation: a cost effective strategy for managed care. Introduction. PMID- 10688943 TI - Prenatal smoking intervention in managed care settings: the Kaiser Permanente Southern California prenatal smoking project. PMID- 10688944 TI - Counselling smokers in Medicaid maternity care: the SCRIPT project. PMID- 10688945 TI - Pre- and postnatal smoking intervention in managed care settings. PMID- 10688946 TI - Prenatal smoking cessation strategies in managed care. PMID- 10688947 TI - Minnesota Decides: a community blueprint for tobacco reduction. PMID- 10688948 TI - Strategic partnerships for addressing tobacco use. PMID- 10688949 TI - Addressing Tobacco in Managed Care program. PMID- 10688950 TI - Smoking cessation clinical practice guideline update and Agency for Healthcare Research and Quality tobacco resources. PMID- 10688951 TI - Resources on tobacco prevention and control available to managed care organisations from the Centers for Disease Control and Prevention. PMID- 10688952 TI - Tobacco control research in managed care: opportunities at the National Cancer Institute. PMID- 10688953 TI - Editorial PMID- 10688954 TI - Lumbar segmental 'instability': clinical presentation and specific stabilizing exercise management. AB - Lumbar segmental instability is considered to represent a significant sub-group within the chronic low back pain population. This condition has a unique clinical presentation that displays its symptoms and movement dysfunction within the neutral zone of the motion segment. The loosening of the motion segment secondary to injury and associated dysfunction of the local muscle system renders it biomechanically vulnerable in the neutral zone. The clinical diagnosis of this chronic low back pain condition is based on the report of pain and the observation of movement dysfunction within the neutral zone and the associated finding of excessive intervertebral motion at the symptomatic level. Four different clinical patterns are described based on the directional nature of the injury and the manifestation of the patient's symptoms and motor dysfunction. A specific stabilizing exercise intervention based on a motor learning model is proposed and evidence for the efficacy of the approach provided. PMID- 10688955 TI - Inter-examiner and intra-examiner agreement for assessing sacroiliac anatomical landmarks using palpation and observation: pilot study. AB - Despite the paucity of research into the reliability of static palpation, it is still employed extensively as a diagnostic tool by manual medicine practitioners. This study tested the inter- and intra-examiner agreement of ten senior osteopathic students using static palpation on ten asymptomatic subjects. Four assessments of the posterior superior iliac spine (PSIS), sacral sulcus (SS), and the sacral inferior lateral angle (SILA) on every subject by all examiners resulted in 1200 assessments in total. Kappa (Kg) yielded intra-examiner agreement that ranged between less-than-chance to substantial for the SILA (Kg= 0.05 to 0.69; mean Kg=0.21), and slight to moderate for the PSIS (Kg=0.07 to 0.58; mean Kg=0.33) and the SS (Kg=0.02 to Kg=0.60; mean Kg=0.24), with 50% significant beyond the 0.05 level. Inter-examiner agreement was slight (PSIS Kg=0.04; SILA Kg=0.08; SS Kg=0.07) and significant at the 0.01 level. Intra examiner agreement was greater than inter-examiner agreement, which was consistent with existing palpation reliability studies. The poor reliability of clinical tests involving palpation may be partially explained by error in landmark location. PMID- 10688956 TI - Measurement of blood flow in the vertebral artery using colour duplex Doppler ultrasound: establishment of the reliability of selected parameters. AB - This study was designed to determine the reliability of the ultrasound testing procedure for evaluating vertebral artery blood flow, and to determine a robust testing protocol for future studies. Blood flow parameters were tested in ten asymptomatic subjects (mean age 33 years, standard deviation 6 years 8 months) using colour duplex Doppler imaging. Volume flow rate data at C5-6 demonstrated good reliability from a single measurement (Intraclass correlation coefficient [ICC]=0.81). Peak velocity sampled at C1-2 showed poor reliability if a single measurement was used (ICC=0.26) improving to fair levels with three measurements (ICC=0.77). Reliability for this parameter was good if five measurements were taken (ICC=0.83-0.84). Systolic/diastolic ratio measured at C5-6 showed poor reliability (ICC=0.57) if a single measurement was taken in the manner of Thiel et al. (1994). This improved to fair reliability (ICC=0.75) if the mean of three measurements was used. There was no further improvement if five measures were sampled. Sampling at C2-3 in the manner of Refshauge (1994) was found to be technically difficult and it was not possible to detect a Doppler shift in three of the ten subjects at this level. Reliability of peak velocity at C2-3 was found to be poor, regardless of whether single or multiple averaged measurements were taken (ICC=0.37-0.63). Mean (time averaged) velocity measurements at C2-3 showed poor reliability if a single measurement was taken (ICC=0.39), fair reliability if the first three measurements were averaged (ICC=0.73) and good to high reliability levels if five measurements were sampled (ICC=0.88-0.91). A review of the literature suggests that sampling volume flow rate at C5-6 and peak velocity at C1-2 represents a clinically meaningful combination of parameters to detect narrowing in the VA. The results of this current study indicate the desirability of taking a single measurement of volume flow rate at C5-6 and the mean of three measurements of peak velocity at C1-2, with the additional calculation of the standard error of measurement, if reliable results are to be achieved. PMID- 10688957 TI - Clinical tests of the sacroiliac joint. A systematic methodological review. Part 1: Reliability. AB - In the literature concerning the sacroiliac joint (SIJ) there are numerous specific tests used to detect joint mobility or pain provocation. In this article the authors have reviewed 11 studies which investigated the reliability of these tests. The methodological quality of the studies was tested by a list of criteria developed by the authors. This list consisted of three categories: (1) study population, (2) test procedures and (3) test results. To each criterion a weighting was attached. The methodological score for nine out of the 11 studies was found to be acceptable. The results of this review, however, could not demonstrate reliable outcomes and therefore no evidence on which to base acceptance of mobility tests of the SIJ into daily clinical practice. There are no indications that 'upgrading' of methodological quality would have improved the final conclusions. With respect to pain provocation tests, the findings did not show the same trend. Two studies demonstrated reliable results using the Gaenslen test and the Thigh thrust test. One study showed acceptable reliability for five other pain provocation tests; however, since other authors have described contradictory results, there is a necessity for further research in this area with an emphasis on multiple test scores and pain provocation tests of the SIJ. PMID- 10688958 TI - Professional issue. Guidance for pre-manipulative testing of the cervical spine. PMID- 10688959 TI - Case report. Clinical instability of the upper cervical spine. PMID- 10688960 TI - Bibliography PMID- 10688961 TI - Hypothalamic neuropeptide Y levels in weaning offspring of low-protein malnourished mother rats. 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. Hypothalamic neuropeptides are decisively involved in the central nervous regulation of body weight and metabolism. We investigated neuropeptide Y (NPY) in distinct hypothalamic nuclei in the offspring of LP mother rats at the end of the critical hypothalamic differentiation period (20th day of life). Weanling LP offspring were underweight (P< 0.001) and hypoinsulinaemic (P< 0.05), while leptin levels were unchanged. NPY was significantly increased in the paraventricular hypothalamic nucleus (PVN) (P< 0.01) and lateral hypothalamic area (P< 0.05) in LP offspring. In contrast, NPY was unchanged in the ventromedial hypothalamic nucleus (VMN). These observations indicate a leptin-independent stimulation of the orexigenic ARC-PVN axis in undernourished LP rats at weaning. Furthermore a disturbed NPYergic regulation of the VMN is suggested, possibly contributing to alterations of the hypothalamic regulation of body weight and metabolism in LP offspring during life. PMID- 10688962 TI - Cerebellin stimulates the secretory activity of the rat adrenal gland: in vitro and in vivo studies. AB - Cerebellin is a 16-aminoacid peptide widely distributed in the central nervous system, where it exerts neuromodulatory functions. Cerebellin is contained in human adrenal medulla, and it has been recently demonstrated that cerebellin elicits catecholamine release by human adrenal in vitro. Aim of the present study was to ascertain whether cerebellin affects adrenal function in the rat. Cerebellin concentration-dependently (from 10(-9)to 10(-7)M) increased norepinephrine (but not epinephrine) and cyclic-AMP production by adrenomedullary tissue in vitro. The norepinephrine response to 10(-7)M cerebellin was blocked by the protein kinase (PK) A inhibitor H-89, but not by the phospholipase C inhibitor U-73122 or the PKC inhibitor calphostin-C. Cerebellin did not affect aldosterone and corticosterone secretion of dispersed zona glomerulosa and zona fasciculata-reticularis adrenocortical cells. Cerebellin concentration dependently (from 10(-8)to 10(-7)M) enhanced norepinephrine release by in situ perfused rat adrenals. Cerebellin (10(-7)M) also elicited a significant rise in aldosterone and corticosterone output, and this effect was annulled by either the beta1-adrenoceptor antagonist l -alprenolol or H-89. Collectively, the present findings allow us to conclude that cerebellin 1) directly stimulates norepinephrine release via the adenylate cyclase/PKA-dependent signaling pathway; and 2) indirectly enhances adrenocortical secretion in vivo, through a paracrine mechanism involving medullary catecholamine release. PMID- 10688963 TI - Tachykinins play a minor role in mediating the third phase of the contractile response to vagal nerve stimulation of the guinea-pig oesophagus. AB - The aim of this study was to determine whether tachykinin receptors might be involved in the mediation of the atropine- and capsaicin-sensitive third phase of a triphasic contractile response to vagal nerve stimulation of the guinea-pig isolated oesophagus. The third phase was inhibited 23.3 +/- 1.7% (P< 0.001, n = 5) and 30. 8 +/- 9.0% (P< 0.05, n = 5) by the NK(3)receptor antagonist, SR 142 801 (0.1 and 1 microM respectively). SR 142 801 (0.1 and 1 microM) had no significant effect on the response to a submaximal concentration of acetylcholine (0.1 mM, n = 4). The third phase was not significantly affected by NK(1)or NK(2)receptor antagonists. Thus, in the guinea-pig oesophagus, it appears that while NK(1)and NK(2)receptors are not involved, NK(3)receptors play a minor role in mediating a contractile response when afferent neurones are excited by vagal nerve stimulation. PMID- 10688964 TI - Cerebrospinal fluid somatostatin levels in febrile seizures and epilepsy in children. AB - We analysed the level of cerebrospinal fluid (CSF) somatostatin in children with febrile seizures and epilepsy. In the febrile seizure group (n = 23), the somatostatin level was 83.9 +/- 11.2 pg/ml, which was significantly higher than that of age-matched controls. CSF samples obtained within 3 h of the last seizure had higher somatostatin levels (106.1 +/- 12.4 pg/ml;n = 14) than did the CSF obtained after 3 h (49.4 +/- 15.6 pg/ml;n = 9). The mean somatostatin level in the epilepsy group was 35.3 +/- 4.3 pg/ml (n = 34), and was distributed as follows: 27.6 +/- 3.6 pg/ml in the idiopathic generalized epilepsy group (n = 16), 44.0 +/- 9.4 pg/ml in the symptomatic generalized epilepsy group (n = 13), and 37.2 +/- 10.1 pg/ml in the partial epilepsy group (n = 5). The levels in each group were significantly higher than those in age-matched controls. Somatostatin is a hypothalamic tetradecapeptide with excitatory effects on neurons in children with febrile seizures and epilepsy. The finding that patients with convulsive disease had elevated levels of CSF somatostatin suggests that somatostatin release is somehow related to seizure activity. It remains to be determined whether this is due to increased release from over-active excitatory neurons or leakage from damaged or anoxic neurons, secondary to seizure activity. PMID- 10688965 TI - Role of third intracellular loop of galanin receptor type 1 in signal transduction. AB - To determine the domains essential for G-protein coupling of the human galanin receptor type 1 (GalR1), we have used both GalR1 mutants and synthetic receptor derived peptides in(125)I-galanin and [(35)S]-GTPgammaS binding studies. Replacement of potential phosphorylation sites by Leu in the third intracellular loop (IC3) of GalR1 did not affect K(D)values for the receptor. Peptides derived form the IC3 loop, and especially the N-terminal part of it were able to increase the rate of [(35)S]-GTPgammaS binding to the trimeric Gialpha1beta1gamma2, but not to Gsalphabeta1gamma2, whereas the peptides corresponding to the IC1 and IC2 loops had no such effect. IC3 loop peptides also inhibited the binding of(125)I galanin to GalR1 in membranes from Rin m5F cells. Our results suggest that the IC3 loop of GalR1, especially its N-terminal part, defines the coupling of the receptor to the Gialpha1beta1gamma2 protein and consequently, to the signal transduction cascade. PMID- 10688966 TI - Peripheral distribution and gene expression of adrenomedullin in the rat: possible source of blood adrenomedullin. AB - Adrenomedullin (AM) was detected in all tissues examined with the highest concentrations in adrenal gland, lung and cardiac atrium. High concentrations of pre-proadrenomedullin mRNA were also detected in the lung, cardiac atrium, adrenal gland, thoracic aorta and mesenteric artery for the first time by solution-hybridization-RNase protection assay. The molecular forms of rat AM in various tissues and plasma were also characterized by Biogel P(30)gel filtration chromatography. We found no significant difference in immunoreactive AM levels between the veins draining the kidney, the lung and the adrenal and the systemic arterial blood. The very low peptide/mRNA ratio and the AM/precursor ratio in the mesenteric artery and thoracic aorta suggest that blood vessels may be the main source of plasma AM. PMID- 10688967 TI - Effects of TRH on acoustic startle, conditioned fear and active avoidance in rats. AB - The effects of intracerebroventricular injection of thyrotropine-releasing hormone (TRH) on acoustic startle, conditioned fear and active avoidance were examined in rats. Acoustic startle was significantly depressed by 12.5 microg TRH, while increasing motor activity. In a fear-potentiated startle paradigm, 12.5 microg TRH reduced the overall startle response amplitude, but did not decrease the amount of fear-potentiated startle. When TRH was administered 15 min before contextual fear conditioning, neither fear-related freezing in acquisition nor in a retention test was affected. In contrast, when TRH was administered 15 min before the retention test, TRH significantly reduced mean percentage of time spent freezing. TRH had no effect on active avoidance. The results demonstrate that TRH decreased acoustic startle and freezing responses, but had little effect on fear conditioning and active avoidance. It is suggested that the results may be due to TRH's effects on motor activity and arousal, independent of its effects on fear. PMID- 10688968 TI - Tachykinin activation of human monocytes from patients with interstitial lung disease, healthy smokers or healthy volunteers. AB - Three types of tachykinin receptors, NK(1), NK(2)and NK(3), have been described to preferentially interact with substance P (SP), neurokinin A (NKA) and neurokinin B (NKB) respectively. Experimental evidence indicates that SP and NKA modulate the activity of inflammatory and immune cells, including mononuclear ones, and points to their involvement in lung pathophysiology. We previously reported that NK(1)and NK(2)receptors are present on monocytes (MO) isolated from healthy donors or rheumatoid patients - a greater sensitivity to NK(2)receptor stimulation was observed in the latter condition. This study evaluated the effects of SP and NKA, as well as NK(1)and NK(2)selective agonists and antagonists, on MO obtained from healthy volunteers, healthy smokers or patients with interstitial lung diseases (e.g. sarcoidosis and idiopathic pulmonary fibrosis). Superoxide anion (O(2)(-)) production was chosen as a parameter of cell activation. SP and NKA dose-dependently evoked O(2)(-)production from MO in all the conditions evaluated, their effects being competitively antagonized by selective antagonists (CP 96 345 and MEN 10 627, respectively). When selective NK(1)and NK(2)agonists were used, [Sar(9)Met(O(2))(11)]SP, a selective NK(1)agonist, induced a more than doubled O(2)production in MO obtained from patients with interstitial lung diseases as compared to healthy volunteers, whereas MO isolated from healthy volunteers were more sensitive to NK(2)receptor stimulation. PMID- 10688969 TI - Modulation of the growth hormone-releasing activity of thyrotropin-releasing hormone in the chicken by its gene-related peptide preproTRH((160-169))(Ps4): enhanced somatostatinergic tone? AB - Recent research demonstrated that endocrine actions of thyrotropin (TSH) releasing hormone (TRH) are modulated by gene-related products within proTRH. In the present report we show that the growth hormone (GH) response to TRH is clearly inhibited after the preincubation of chicken pituitary glands with preproTRH((160-169))Ps4, whereas the TSH response is not impaired. Binding sites for(125)I-[Tyr(0)]-Ps4 were, however, not detected on chicken pituitary membranes, although (as a control) they were readily detectable on membranes from rat pituitary glands. An indirect action may therefore take place within the pituitary by modulating the action of somatostatin (SRIH), the inhibitor of GH release in the chicken. This hypothesis is strengthened by the observation that Ps4 increases the binding of(125)I-[Tyr(1)]-SRIH to chicken pituitary membranes in a dose-related way. Since Ps4 is also produced by pituitary tissue, this may reflect a local or paracrine action on the regulation of GH release. PMID- 10688970 TI - Neuropeptide Y attenuates the effect of locus coeruleus denervation by DSP-4 treatment on social behaviour in the rat. AB - Noradrenaline (NA) has been implicated in both increase and reduction of anxiety. Selective destruction of nerve endings of the locus coeruleus projections by DSP 4 has been shown to reduce active behaviour in novel situations by enhancing anxiety. In the present study, DSP-4 (50 mg/kg) treatment reduced locomotor activity and time spent in social interaction in rats placed into a novel environment together with an unfamiliar rat, indicating an anxiogenic-like effect. The effect of DSP-4 on time spent in social interaction was completely antagonized by intracerebroventricular administration of neuropeptide Y (NPY) (1 microg) which had no effect of its own on this measure. The present study thus supports the idea that DSP-4 pretreatment is anxiogenic in novel situations and suggests a functional relationship of NA- and NPY-using neural mechanisms in the regulation of social behaviour. PMID- 10688971 TI - Modulatory effect of endogenous and exogenous opioids on the excitatory reflex pathway of the rat ileum. AB - The ascending excitatory reflex is part of the peristaltic reflex, an important participant in intestinal propulsion. The aim of this study was to characterize the role of different opioid receptors in the ascending reflex through exogenous application of non-selective (Met-enkephalin) and selective opioid agonists (mu PLO17, delta-DPDPE, kappa-U-50, 488) as well as selective opioid receptor antagonists (mu: CTOP-NH(2), delta: ICI-174,864, kappa: Nor-Binaltorphimine). Metenkephalin (IC(50): 0.06 microM) and morphine (IC(50): 1.8 microM) inhibited the ascending reflex response concentration-dependently. Both the mu-selective agonist PLO17 (IC(50): 0.83 microM, n =11) and the kappa-selective agonist U 50,488 (IC(50): 0.68 microM, n =8) concentration-dependently inhibited the magnitude of the ascending contractile reflex response, whereas the delta-agonist DPDPE (10(-10)-10(-6)M) had no significant effect. In contrast, the latency of the response (time interval between start of the stimulus and onset of the contraction) was significantly prolonged by PLO17 > morphine > Met-enkephalin > DPDPE, whereas U-50,488 showed no effect. When the effect of the receptor specific antagonists was tested, only CTOP-NH(2)and Nor-BNI caused a significant increase of the contractile response, whereas ICI-174 864 was ineffective. On the other hand, CTOP-NH(2)> ICI-174 864 decreased the latency significantly but the kappa-receptor agonist Nor-BNI had no influence. Thus, mu- and kappa-receptors seem to be involved in regulating the contraction strength of the ascending reflex, whereas both mu- and delta-receptors seem to be involved in the timing of the reflex response. PMID- 10688972 TI - Mu and delta opioid receptor regulation of pro-opiomelanocortin peptide secretion from the rat neurointermediate pituitary in vitro. AB - We investigated the ability of selective opioid agonists and antagonists to influence pro-opiomelanocortin peptide secretion from the rat neurointermediate lobe in vitro. The mu-opioid agonist DAMGO ([D-Ala(2), N-Me-Phe(4), Gly(5) ol]enkephalin) significantly stimulated beta-endorphin and alpha-melanocyte stimulating hormone release relative to controls early (30 min) in the incubation period. Similar effects on beta-endorphin secretion were observed with the selective mu-opioid agonist dermorphin. The delta-opioid receptor agonist DPDPE ([D-Pen(2,5)]enkephalin) weakly inhibited beta-endorphin secretion relative to controls while the kappa-opioid receptor agonist U50488 had no effect. The mu opioid selective antagonist CTOP (D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2)) inhibited basal beta-endorphin secretion while kappa- and delta-opioid receptor antagonists had no effect. Our data support a role for local mu-opioid receptor control of intermediate lobe pro-opiomelanocortin peptide secretion. Peptide secretion from melanotropes appears to be tonically stimulated by activation of mu-opioid receptors in the absence of intact neuronal innervation to the intermediate lobe. PMID- 10688973 TI - Opioid activity profiles indicate similarities between the nociceptin/orphanin FQ and opioid receptors. AB - Nociceptin (orphanin FQ) is the recently discovered peptide agonist for the orphan receptor opioid receptor-like 1 (ORL1). Despite the high sequence homology between ORL1 and the opioid receptors, most opioids lack affinity for the nociceptin receptor. The affinity and functional profile of opioids possessing activity at the nociceptin receptor was determined using [3H]nociceptin and nociceptin-stimulated [35S]GTPgammaS binding. The mu-opioid receptor-selective agonist lofentanil potently and competitively displaced [3H]nociceptin at rat brain receptors (IC(50) 62 nM). Lofentanil exhibited full agonism for enhancement of [35S]GTPgammaS binding to human recombinant ORL1 receptors (EC(50) 50 nM). The related piperidines ohmefentanyl and sufentanil and the nonselective opioid receptor agonist etorphine were less potent nociceptin receptor agonists. The kappa(1)+kappa(3)-opioid receptor agonist/mu-opioid receptor antagonist naloxone benzoylhydrazone was a pure antagonist at both rat brain and human ORL1 receptors. The nonselective opioid receptor partial agonist buprenorphine and the nonselective opioid receptor antagonist (-)-quadazocine exhibited pure antagonism at rat brain receptors, but displayed partial agonism at human ORL1 receptors. Thus, opioids displaying full agonism at the nociceptin receptor are also opioid receptor agonists, whereas opioids that are antagonists or partial agonists at the nociceptin receptor show antagonism or partial agonism at opioid receptors. In addition, the stereospecificity required at opioid receptors appears to be retained at the nociceptin receptor, since (+)-quadazocine is inactive at both receptors. These findings illustrate the structural and functional homology of the opioid recognition site on these two receptor classes and suggest that opioids may provide leads for the design of nonpeptide nociceptin receptor agonists and antagonists lacking affinity for the classical opioid receptors. PMID- 10688974 TI - Essential role of extracellular charged residues of the human CCK(1) receptor for interactions with SR 146131, SR 27897 and CCK-8S. AB - We hypothesized that charge-charge interactions may be important for the binding of the human cholecystokinin type 1 (CCK(1)) receptor-specific non-peptide full agonist SR 146131, (2-[4-(4-chloro-2, 5-dimethoxyphenyl)-5-(2-cyclohexyl-ethyl) thiazol-2-ylcarbamoyl ]-5, 7-dimethyl-indol-1-yl-1-acetic acid), the competitive antagonist SR 27897, (1-[2-(4-(2-chlorophenyl)thiazol-2-yl) aminocarbonyl indoyl] acetic acid) and the natural octapeptide CCK-8S to the CCK(1) receptor. Alanine replacement studies of positively charged residues in the extracellular domains of the receptor showed that only the R336A mutation affected SR 146131 potency of mutated receptors transiently expressed in monkey kidney epithelial COS-7 cells. Two residues, Lys(115) and Lys(187), were implicated in SR 27897 binding. Only the replacement of Lys(115), Arg(197) and Arg(336) significantly affected CCK-8S binding or activity. These results clearly indicated the importance of certain charged residues, but not others, in SR 146131, SR 27897 and CCK-8S binding. Furthermore, although these molecules probably occupy different binding sites on the CCK(1) receptor, we show that a small non-peptide agonist, SR 146131, can stimulate the dual signaling pathways mediated by the CCK(1) receptor. PMID- 10688975 TI - Activation of multiple mitogen-activated protein kinases by recombinant calcitonin gene-related peptide receptor. AB - Calcitonin gene-related peptide is a 37-amino-acid neuropeptide and a potent vasodilator. Although calcitonin gene-related peptide has been shown to have a number of effects in a variety of systems, the mechanisms of action and the intracellular signaling pathways, especially the regulation of mitogen-activated protien kinase (MAPK) pathway, is not known. In the present study we investigated the role of calcitonin gene-related peptide in the regulation of MAPKs in human embryonic kidney (HEK) 293 cells stably transfected with a recombinant porcine calcitonin gene-related peptide-1 receptor. Calcitonin gene-related peptide caused a significant dose-dependent increase in cAMP response and the effect was inhibited by calcitonin gene-related peptide(8-37), the calcitonin gene-related peptide-receptor antagonist. Calcitonin gene-related peptide also caused a time- and concentration-dependent increase in extracellular signal-regulated kinase (ERK) and P38 mitogen-activated protein kinase (P38 MAPK) activities, with apparently no significant change in cjun-N-terminal kinase (JNK) activity. Forskolin, a direct activator of adenylyl cyclase also stimulated ERK and P38 activities in these cells suggesting the invovement of cAMP in this process. Calcitonin gene-related peptide-stimulated ERK and P38 MAPK activities were inhibited significantly by calcitonin gene-related peptide receptor antagonist, calcitonin gene-related peptide-(8-37) suggesting the involvement of calcitonin gene-related peptide-1 receptor. Preincubation of the cells with the cAMP dependent protein kinase inhibitor, H89 [?N-[2-((p-bromocinnamyl)amino)ethyl]-5 isoquinolinesulfonamide, hydrochloride?] inhibited calcitonin gene-related peptide-mediated activation of ERK and p38 kinases. On the other hand, preincubation of the cells with wortmannin ?[1S-(1alpha,6balpha,9abeta,11alpha, 11bbeta)]-11-(acetyloxy)-1,6b,7,8,9a,10,11, 11b-octahydro-1-(methoxymethyl) 9a,11b-dimethyl-3H-furo[4,3, 2-de]indeno[4,5-h]-2-benzopyran-3,6,9-trione?, a PI3 kinase inhibitor, attenuated only calcitonin gene-related peptide-induced ERK and not P38 MAPK activation. Thus, these data suggest that activation of ERK by calcitonin gene-related peptide involves a H89-sensitive protein kinase A and a wortmannin-sensitive PI3-kinase while activation of p38 MAPK by calcitonin gene related peptide involves only the H89 sensitive pathway and is independent of PI3 kinase. This also suggests that although both ERK and P38 can be activated by protein kinase A, the distal signaling components to protein kinase A in the activation of these two kinases (ERK and P38) are different. PMID- 10688976 TI - The effect of protein kinase C activation on colonic epithelial cellular integrity. AB - We have investigated whether activation of protein kinase C has a direct cytotoxic effect on colonic mucosal epithelial cells and whether oxidant-induced damage to colonocytes is mediated by activation of cellular protein kinase C. Incubation of freshly harvested cells from rat colon with the protein kinase C activator, phorbol 12-myristate, resulted in a concentration-dependent increase in the extent of cell injury. Phorbol 12-myristate acetate (0.1-10 microM) also increased cellular protein kinase C activity and this was reduced significantly by treating cells with the antagonists staurosporine or 2-[1-(3 dimethylaminopropyl)-indol-3-yl]3-(-indol-3-yl)maleimide (GF 109203X; 10 microM). Phorbol 12-myristate acetate treatment also resulted in increased translocation of proteins for protein kinase C isoforms alpha, delta and epsilon from cytosol to membrane particulate fractions. The antagonists reduced the extent of cell damage in response to phorbol 12-myristate acetate. Furthermore, cell injury in response to the phorbol acetate was also inhibited by the addition of the oxidant scavengers, superoxide dismutase or catalase to the cell suspension. Addition of H(2)O(2) to the incubation medium (0.1-100 microM) resulted in an increase in cellular protein kinase C activity, an increase in the expression of the alpha, beta and zeta isoforms and a reduction in cell integrity. The cellular damaging actions of H(2)O(2) were significantly reduced by the protein kinase C antagonists, staurosporine or 2-[1-(3-dimethylaminopropyl)-indol-3-yl]-3-(-indol 3-yl)maleimide (GF 109203X). These findings suggest that protein kinase C activation results in colonic cellular injury and this damage is mediated, at least in part, by release of reactive oxidants. Furthermore, oxidant-mediated damage to these cells also involves protein kinase C activation. PMID- 10688977 TI - Cellular uptake and interaction with purified membranes of rebeccamycin derivatives. AB - Rebeccamycin is an antitumor antibiotic possessing a DNA-intercalating indolocarbazole chromophore linked to a glycosyl residue. The carbohydrate moiety of rebeccamycin and related synthetic analogues, such as the potent antitumor drug NB-506 (6-N-formylamino-12,13-dihydro-1, 11-dihydroxy-13-(beta-D glucopyranosyl)-5H-indolo[2,3-a]pyrrolo- [3,4-c]carbazole-5,7-(6H)-dione), is a key element for both DNA-binding and inhibition of DNA topoisomerase I. In this study, we have investigated the cellular uptake of rebeccamycin derivatives and their interaction with purified membranes. The transport of radiolabeled [3H]dechlorinated rebeccamycin was studied using the human leukemia HL60 and melanoma B16 cell lines as well as two murine leukemia cell lines sensitive (P388) or resistant (P388CPT5) to camptothecin. In all cases, the uptake is rapid but limited to about 6% of the drug molecules. In HL60 cells, the uptake entered a steady-state phase of intracellular accumulation of about 0.26+/-0.05 pmol/10(6) cells, which persisted to at least 90 min. The efflux of exchangeable radiolabeled molecules was relatively weak. Fluorescence studies were performed to compare the interaction of a rebeccamycin derivative and its aglycone with membranes purified from HL60 cells. The glycosylated drug molecules bound to the cell membranes can be extracted upon washing with buffer or by adding an excess of DNA. In contrast, the indolocarbazole drug lacking the carbohydrate domain remains tightly bound to the membranes with very little or no exchange upon the addition of DNA. The membrane transport and binding properties of indolocarbazole drugs related to rebeccamycin are reminiscent to those of other DNA-intercalating antitumor agents. The uptake most likely occurs via a passive diffusion through the plasma membranes and the glycosyl residue of the drug plays an essential role for the translocation of the drug from the membranes to the internal cell components, such as DNA. PMID- 10688978 TI - Region-specific changes in 5-HT(1A) receptor-activated G-proteins in rat brain following chronic buspirone. AB - 5-Hydroxytryptamine(1A) (5-HT(1A)) receptors, which activate inhibitory G proteins, are implicated in psychiatric disorders including anxiety and depression. Studies suggest that chronic 5-HT(1A) receptor agonist administration alters 5-HT(1A) receptor function, but the effect of chronic treatment on 5 HT(1A) receptor-activated G-proteins is unclear. In this study, agonist stimulated [35S]guanylyl-5'-O-(gamma-thio)-triphosphate (GTPgammaS) binding was examined following chronic administration of buspirone. Brains were processed for [35S]GTPgammaS autoradiography using R(+)-8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) for 5-HT(1A) receptors or baclofen for GABA(B) receptors. Net 8-OH DPAT-stimulated [35S]GTPgammaS binding was decreased by 25-30% in the septum and dorsal raphe nucleus of buspirone-treated animals. No significant changes in 8-OH DPAT-stimulated [35S]GTPgammaS binding were found in the prefrontal, entorhinal or cingulate cortices or hippocampus in buspirone-treated rats. GABA(B) receptor stimulated [35S]GTPgammaS binding was increased by 25% in the hippocampus, with no significant changes in any other region examined. These results demonstrate region-specific alterations in 5-HT(1A) and GABA(B) receptor-activated G-proteins following chronic buspirone treatment, which may contribute to the clinical effects of this drug. PMID- 10688979 TI - 5-hydroxytryptamine interaction with the nicotinic acetylcholine receptor. AB - The present study examines the interaction of the neurotransmitter 5 hydroxytryptamine (5-HT) with muscle-type nicotinic acetylcholine receptors. 5-HT inhibits the initial rate of [125I]alpha-bungarotoxin binding to Torpedo acetylcholine receptor membranes (IC(50)=8.5+/-0.32 mM) and [3H]5-HT can be photoincorporated into acetylcholine receptor subunits, with labeling of the alpha-subunit inhibitable by both agonists and competitive antagonists. Within the agonist-binding domain, [3H]5-HT photoincorporates into alphaTyr(190), alphaCys(192) and alphaCys(193). Functional studies using the human clonal cell line TE671/RD, show that 5-HT is a weak inhibitor (IC(50)=1.55+/-0.25 mM) of acetylcholine receptor activity. In this regard, agonist-response profiles in the absence and presence of 5-HT indicate a noncompetitive mode of inhibition. In addition, 5-HT displaces high affinity [3H]thienylcyclohexylpiperidine binding to the desensitized Torpedo acetylcholine receptor channel (IC(50)=1.61+/-0.07 mM). Collectively, these results indicate that 5-HT interacts weakly with the agonist recognition site and inhibits receptor function noncompetitively by binding to the acetylcholine receptor channel. PMID- 10688980 TI - Tolerance to morphine at the mu-opioid receptor differentially induced by cAMP dependent protein kinase activation and morphine. AB - Human neuroblastoma SH-SY5Y cells express endogenous mu-opioid receptor and develop cellular tolerance to morphine after prolonged (>/=4 h) treatment with morphine. Treatment with forskolin (25 microM, 12 h), an adenylyl cyclase activator, also desensitized mu-opioid receptor response to morphine (10 microM) by 38% (P<0. 001), which was reversed by the cyclic AMP (cAMP) dependent kinase inhibitor N-(2-aminoethyl)-5-isoquinolinesulfonamide (H8) (100 microM). Treatment with both morphine and forskolin appeared to cause an additive effect in desensitizing mu-opioid receptor. In mu-opioid receptor stably transfected human embryonic kidney 293 (HEK-mu) cells, morphine treatment produced cAMP upregulation, yet failed to induce mu-opioid receptor tolerance. However, treatment with forskolin (25 microM) or 8-bromo-cAMP (1mM) led to profound mu opioid receptor tolerance, which was reversed by H8. These results demonstrate that cAMP-dependent kinase activation causes mu-opioid receptor tolerance. However, morphine-induced mu-opioid receptor tolerance in SH-SY5Y cells is not mediated by cAMP-dependent kinase activation. In addition, our results indicate that cAMP-upregulation does not necessarily lead to mu-opioid receptor tolerance. PMID- 10688981 TI - Inhibitory effect of orally administered donepezil hydrochloride (E2020), a novel treatment for Alzheimer's disease, on cholinesterase activity in rats. AB - Donepezil hydrochloride ((+/-)-2-[(1-benzylpiperidin-4-yl)methyl]-5, 6-dimethoxy indan-1-one monohydrochloride: E2020: donepezil) is a potent and selective acetylcholinesterase inhibitor developed for the treatment of Alzheimer's disease. The present experiments were designed to compare the inhibitory effects of orally administered donepezil and other cholinesterase inhibitors, tacrine (9 amino-1,2, 3,4-tetrahydroacridine hydrochloride), (S)-N-ethyl-3-[(1-dimethyl amino)ethyl]-N-methyl-phenylcarbamate hydrogentartrate (ENA-713, rivastigmine) and 3-[1-(phenylmethyl)-4-piperidinyl]-1-(2,3,4, 5-tetrahydro-1H-1-benzazepin-8 yl)-1-propanone fumarate (TAK-147), on the cholinesterase activity in the brain and plasma of rats. Moreover, in order to validate the cholinesterase inhibition data, we measured the brain and plasma concentrations of these drugs. Oral administration of donepezil, tacrine, ENA-713 or TAK-147, caused a dose-dependent inhibition of brain and plasma cholinesterase activities. The ID(50) values of these compounds for brain cholinesterase activity were 6.3, 40.5, 7.2 and 26.8 micromol/kg, respectively. On the other hand, the ID(50)170, 9.7 and 51.2 micromol/kg, respectively. Thus, the ratios of the ID(50)4.2, 1.3 and 1.9, respectively. Brain and plasma concentrations of donepezil, tacrine and TAK-147 increased dose-dependently. The ratios of the concentrations (brain/plasma) of these compounds were 6.1-8.4 for donepezil, 14.5-54.6 for tacrine and 7.0-20.6 for TAK-147. The values of 50% inhibitory concentration of these drugs in the brain were 0.42, 3.5 and 1.1 nmol/g, respectively. In contrast, the brain and plasma concentrations of ENA-713 at all doses, except the two highest doses, were below the quantification limit. These results suggest that orally administered donepezil satisfactorily penetrates into the brain and inhibits cholinesterase there, and that donepezil is a potent and selective inhibitor of brain cholinesterase in comparison with plasma cholinesterase in vivo. PMID- 10688982 TI - Cross-tolerance between analgesia produced by xylazine and selective opioid receptor subtype treatments. AB - Opioid receptor agonists produce analgesia through multiple systems activated by stimulation of mu(1), mu(2), delta(1), delta(2) and kappa(1) opioid receptors. Morphine analgesia is modulated by stimulation of alpha(2) adrenoceptors. To understand how multiple opioid analgesic systems interact with alpha(2) adrenoceptor systems, analgesic cross-tolerance between the alpha(2) adrenoceptor agonist xylazine and opioid receptor agonists was studied using the mouse tail flick assay. Mice received either xylazine (20 mg/kg, s.c.) or saline (1 ml/kg) for five days. On day six, mice received a dose of s.c. xylazine, i.c.v. [D Ala(2),MePhe(4),Gly(ol)(5)]enkephalin (DAMGO), i.t. Tyr-Pro-Trp-Gly-NH(2) (Tyr-W MIF-1), i.c.v. or i.t. [D-Pen(2),D-Pen(5)]enkephalin (DPDPE), i.t. [D Ala(2)]deltorphin II (deltorphin II), or s.c. trans-(+/-)-3, 4-dichloro-N-methyl N-[2-(1-pyrrolidinyl-cyclohexyl] benzeneacetamide (U50,488). Xylazine tolerant mice required 4. 57-fold more xylazine to elicit the same response as saline treated animals and showed a 2.55-fold shift in i.c.v. DAMGO and a 3.37-fold shift in i.c.v. DPDPE antinociception. No cross-tolerance was seen with i.c.v. deltorphin II, i.t.Tyr-W-MIF-1, i.t. DPDPE, i.t. Tyr-W-MIF-1 or s.c. U50,488. These results implicate alpha(2) adrenoceptor systems in the modulation of supraspinal mu(1), and delta(1) opioid analgesic circuitry and raise the possibility that mu(2), delta(2) or kappa(1) opioid receptor agonists may be alternated with alpha(2) adrenoceptor agonists to minimize tolerance or treat opioid-tolerant patients. PMID- 10688983 TI - Noradrenergic lesion antagonizes desipramine-induced adaptation of NMDA receptors. AB - Repeated administration of the tricyclic antidepressant, desipramine, for 28 days to mice effected a decrease in the potency of glycine to displace [3H]5,7 dichlorokynurenic acid (5,7-DCKA) in mouse cortical homogenates. Pre-treatment with the noradrenergic neurotoxin DSP-4, while having no effect alone, attenuated the desipramine-induced effect. The present findings support a norepinephrine dependent adaptation of the NMDA receptor complex in vivo following chronic desipramine treatment. The inter-relationship of norepinephrine and glutamate transmission may provide insight into the mechanism underlying the action of antidepressant drugs. PMID- 10688984 TI - Motor effects of (-)-OSU6162 in primates with unilateral 6-hydroxydopamine lesions. AB - The effects of the novel compound, (-)-OSU6162 ((S)-(-)-3-methylsulfonylphenyl-1 propylpiperidine), on rotational behavior induced by dopamine receptor agonists was investigated in common marmosets (Callithrix jacchus) with unilateral 6 hydroxydopamine lesions. (-)-OSU6162 per se displayed no effect on the animals' behavior. On the other hand, pretreatment with (-)-OSU6162 attenuated rotational behavior induced by apomorphine (apomorphini hydrochloridum), L-DOPA (3,4 dihydroxyphenylalanine), and the dopamine D2 receptor agonist, quinpirole (trans (-)-4aR-4,4a, 5,6,7,8,8a,9-octahydro-5-propyl-1H-pyrazolol[3,4-g]quinoline hydrochloride), without inducing motor impairment such as akinesia or dystonia. In addition, treatment with (-)-OSU6162 for 5 consecutive days almost completely abolished the rotational behavior provoked by apomorphine and produced a transient subsensitization of such apomorphine-induced effects after it was discontinued. Moreover, pretreatment with (-)-OSU6162 in two monkeys augmented the rotational behavior elicited by the dopamine D1 receptor agonists, SKF-81297 (R(+)-6-chloro-7,8,dihydroxy-1-phenyl-2,3,4, 5-tetrahydro-1H-3-benzazepine hydrobromide) and A-77636 ((-)-(1R, 3S)-3-adamantyl-1-(aminomethyl)-3,4-dihydro 5, 6-dihydroxy-1H-2-benzopyran hydrochloride). The findings indicate that (-) OSU6162 can exert indirect state-dependent effects that differentially affect dopamine D1 and dopamine D2 receptor agonist-induced behavior. PMID- 10688985 TI - Antithrombotic efficacy of RPR208566, a novel factor Xa inhibitor, in a rat model of carotid artery thrombosis. AB - Coagulation factor Xa is the sole enzyme responsible for activating the zymogen prothrombin to thrombin, resulting in fibrin generation, platelet activation, and subsequent thrombus formation. Our objective was to evaluate the antithrombotic efficacy of the novel factor Xa inhibitor, 2-(3-carbamimidoyl-benzyl)-3-[(3', 4'dimethoxy-biphenyl-4-carbonyl)-amino]-butyric acid methyl ester trifluoroacetate (RPR208566), in a well-established rat model of arterial thrombosis, and to compare the results with those obtained with argatroban and heparin, direct and indirect inhibitors of thrombin, respectively. Thrombus formation was initiated by placing a filter paper saturated with FeCl(2) on the adventia of the carotid artery for 10 min. Time-to-occlusion was measured from initiation of injury until blood flow reached zero. Formed thrombi were removed and weighed 60 min after the placement of the filter paper. RPR208566, heparin, and argatroban dose-dependently increased time-to-occlusion and reduced thrombus mass. When administered at 500 microgram/kg+50 microgram/kg/min, RPR208566 prolonged time-to-occlusion to 56+/-4 min (vs. 18+/-2 min for vehicle) and reduced thrombus mass to 3.0+/-0.7 mg (vs. 7.3+/-0.6 mg for vehicle). The highest doses of argatroban (500 microgram/kg+50 microgram/kg/min) and heparin (300 U/kg+10 U/kg/min) increased time-to-occlusion to the maximum of 60 min and decreased thrombus mass to 5.5+/-0.8 and 2.6+/-0.3, respectively. The antithrombotic effects of heparin and argatroban at these doses were associated with increases in activated partial thromboplastin time of 5.6+/-0.9- and 2.9+/ 0.3-fold over baseline, respectively. However, the highest dose of RPR208566 produced a modest 1.3+/-0.1-fold increase in activated partial thromboplastin time. These results indicate that factor Xa inhibition with compounds such as RPR208566 may be an attractive mechanism for novel antithrombotic drug therapy. PMID- 10688986 TI - Effects of nitric oxide-modulating amino acids on coronary vessels: relevance to sepsis. AB - Excessive nitric oxide (NO) production in septic shock is thought to contribute to the associated profound hypotension. Here we show that despite induction of NO synthase (NOS) in the hearts of endotoxin-treated rats, coronary vascular responses to the contractile peptide endothelin-1, were not modified. This was not due to any change in the expression of endothelin receptors. However, when the substrate for NOS, L-arginine, was added to the perfusate, increases in coronary perfusion pressure stimulated by endothelin were reduced in hearts from endotoxin-treated animals compared to those from controls. In addition, L glutamine, which blocks the generation of L-arginine from intracellular stores, enhanced the increase in perfusion pressure stimulated by endothelin-1. These data suggest that L-arginine becomes rate limiting for the production of NO in the coronary vessels during septic shock. Moreover, it suggests that vascular reactivity may be modulated positively or negatively by supplementation with the relevant amino acids. PMID- 10688987 TI - Electrical and mechanical effects of vasoactive intestinal peptide and pituitary adenylate cyclase-activating peptide in the rat colon involve different mechanisms. AB - This work aimed to study the effects of pituitary adenylate cyclase-activating peptide (PACAP) and vasoactive intestinal peptide (VIP) on the mechanical and electrical activity of the circular muscle of the rat colon and the mechanisms involved in such effects. Spontaneous mechanical activity was studied in vitro in an organ bath and the membrane potential was recorded using the microelectrode technique. Both VIP and PACAP (0.1 microM) caused an immediate, sustained and tetrodotoxin (1 microM)-resistant inhibition of the cyclic spontaneous mechanical activity and hyperpolarization. The small-conductance Ca(2+)-activated K(+) channel blocker, apamin (1 microM), did not change the VIP- and PACAP-induced relaxation but reduced the hyperpolarization induced by PACAP whereas it did not change that induced by VIP. In contrast, the purinoceptor antagonist, suramin (100 microM), blocked the hyperpolarization caused by PACAP and VIP but failed to change their mechanical inhibitory effects. Moreover, the putative PACAP and VIP receptor antagonists, PACAP-(6-38) and VIP-(10-28), respectively, both 3 microM, failed to change the effects of either peptide and modified neither the inhibitory junction potential nor the relaxation induced by electrical-field stimulation. Thus, these results suggest that the mechanisms mediating relaxation are not strictly coupled to the mechanisms mediating hyperpolarization. This could be due to activation of two distinct mechanisms of action after agonist receptor interaction. PMID- 10688988 TI - Effects of eicosanoids, neuromediators and bioactive peptides on murine airways. AB - The effects of several mediators including prostanoids, neuromediators, bioactive peptides and leukotrienes were investigated on the trachea, upper bronchi, lower bronchi and lung parenchyma of selected strains of mice mounted in a cascade superfusion system. The upper airways (trachea, upper bronchi) responded with greater maxima than lower airways (lower bronchi, lung parenchyma). Acetylcholine, carbachol, serotonin and 9, 11-dideoxy-9alpha,11alpha-epoxymethano prostaglandin F(2alpha)serotonin>/=acetylcholine. Prostaglandins E(2), F(2alpha) and D(2)90% relaxation in some cases. The rank order of potency for the prostaglandins was E(2)>/=F(2alpha)D(2) with the exception of the lower bronchi on which prostaglandins had the following order of potency: F(2alpha)>/=E(2)D(2). The effects of prostaglandins were similar in four commonly used strains of mice (CD-1, BALB/c, C57BL/c6 and C3H) with some variations in efficacy. Iloprost was a weak mouse airway relaxant. It had the greatest effect on the trachea and bronchi of BALB/c and C57BL/c6 mice, whereas it had little or no effect on the airways of the CD-1 and C3H mouse strains. Vasoactive intestinal peptide potently relaxed the carbachol and precontracted the mouse trachea and bronchi. However, vasopressin, another bioactive peptide, potently and efficaciously contracted the mouse trachea and upper bronchi but had little effect on the lower bronchi. Vasopressin was the most potent and efficacious contractile agonist tested in this study. Contractions were observed with endothelins-1, -2 and -3 on mouse trachea and bronchi, but marked tachyphylaxis was present. Sarafotoxin s6c followed the same pattern suggesting the presence of endothelin ET(B) receptors on the mouse airways. Of all leukotrienes assayed (B(4), C(4), D(4) and E(4)) only leukotriene C(4) weakly contracted the mouse trachea and upper bronchi, but tachyphylaxis was most evident. PMID- 10688989 TI - Effects of mu-opioid receptor agonists on intestinal secretion and permeability during acute intestinal inflammation in mice. AB - We evaluated and compared the effects of mu-opioid receptor agonists on mucosal fluid transport and permeability, during acute intestinal inflammation. We hypothesized that inflammation would sensitize mu-opioid receptors in the submucosal plexus and/or enterocytes enhancing the effects of mu-opioid receptor agonists. Inflammation was induced by intragastric administration of croton oil, whereas controls received saline. Fluid transport was assessed by enteropooling, and intestinal permeability by blood-to-lumen passage of [51Cr] etylenediaminetetraacetate ([51Cr] EDTA). Intestinal inflammation induced a significant increase in enteropooling (1.9 times) and permeability (2.5 times). In saline- and croton oil-treated animals, mu-opioid receptor agonists produced dose-related inhibitions of enteropooling and intestinal permeability. During inflammation, the potency of morphine increased 4.8 and 3.7 times, inhibiting enteropooling and intestinal permeability, respectively; the potencies of fentanyl and PL017 similarly increased by approximately three (enteropooling) and two times (permeability) in croton oil animals. All effects were reversed by naloxone and naloxone methiodide. The results show that inflammation increases the inhibitory potency of mu-opioid receptor agonists on secretion and permeability, suggesting a sensitization of peripheral mu-opioid receptors. PMID- 10688990 TI - Anti-inflammatory glycoterpenoids from Scrophularia auriculata. AB - The activity of the four glycoterpenoids: two saponins, verbascosaponin A and verbascosaponin, and two iridoids, scropolioside A and scrovalentinoside, isolated from Scrophularia auriculata ssp. pseudoauriculata, were studied in different models of acute and chronic inflammation. Both saponins significantly inhibited the mouse paw edema induced by carrageenan and ear edema induced by single and multiple doses of 12-O-tetradecanoylphorbol 13-acetate (TPA). Verbascosaponin A showed a potency twice as high as that of indomethacin in the acute TPA model. Verbascosaponin A and scropolioside A were active after a long latency period against ethyl phenylpropiolate edema, as are glucocorticoids. When the putative corticoid-like mechanism of the two compounds was studied, verbascosaponin A activity was notably reduced by the mRNA synthesis inhibitor, actinomycin D, while the effect of scropolioside A was partially interfered with by the anti-glucocorticoid drugs used. Both iridoids were active on the delayed type hypersensitivity reaction. They significantly reduced the inflammatory lesion and suppressed the cellular infiltration. PMID- 10688991 TI - (R)-ACX is a novel sufonylurea compound with potent, quick and short-lasting hypoglycemic activity. AB - We investigated the mechanism of the hypoglycemic effect of (R)-4-(1 acetoxyethyl)-N-(cyclohexylcarbamoyl)benzene-sulfonamide [(R)-acetoxyhexamide; (R)-ACX], a new sulfonylurea compound. (R)-ACX potently stimulated the release of insulin from cultured pancreatic beta-cells (HIT T15 cells), established from hamster islet cells SV40-transformed. When (R)-ACX was orally administered to fasted rats, it decreased the plasma glucose level in a dose-dependent manner. The hypoglycemic effect of (R)-ACX was quick and short lasting, as compared to that of acetohexamide and glibenclamide. The quick and short-lasting hypoglycemic effect of (R)-ACX was thought likely to result from rapid absorption of (R)-ACX and rapid elimination of (R)-ACX and its metabolite, (R)-hydroxyhexamide. Furthermore, (R)-ACX was found to suppress the increase of blood glucose level due to starch loading in fasted mice. (R)-ACX may be useful in the control of postprandial hyperglycemia to patients with non-insulin-dependent diabetic mellitus. PMID- 10688992 TI - Male factor subfertility: possible causes and the impact of nutritional factors. AB - OBJECTIVE: To review possible causes for male factor subfertility with emphasis on nutritional factors such as zinc and folate. DESIGN: A literature search was performed on MEDLINE and via bibliographies of published works. RESULT(S): Many causes for male factor subfertility are described in the literature. Both environmental and genetic factors could play a role. However, the pathogenesis of male factor infertility is poorly understood, including the role of specific micronutrients such as zinc and folate. Both zinc and folate are involved in the synthesis of DNA and RNA. Despite the fact that zinc deficiency leads to several clinical symptoms such as decreased spermatogenesis and impaired male fertility, the exact pathophysiology has not been clarified. CONCLUSION(S): Because most causes of male factor subfertility are unknown, more research is needed. Because male factor subfertility due to nutritional deficiencies is in principle amenable to curative and/or preventive action by supplementation, emphasis should be put on studies on the effect of specific nutrients on male fertility. PMID- 10688993 TI - Ethical issues in ovarian transplantation and donation. PMID- 10688994 TI - On assisted reproduction, religion, and civil law. PMID- 10688995 TI - Use of frozen-thawed testicular sperm for intracytoplasmic sperm injection. AB - OBJECTIVE: To determine the feasibility of using frozen-thawed testicular spermatozoa for intracytoplasmic sperm injection. DESIGN: Prospective clinical study. SETTING: A university hospital. PATIENT(S): One hundred seventy-five azoospermic men participating in a routine intracytoplasmic sperm injection program. INTERVENTION(S): The men underwent testicular biopsy for cryopreservation of tissue to be used in consecutive intracytoplasmic sperm injection treatment cycles. Their female partners underwent controlled ovarian hyperstimulation for conventional IVF treatment. MAIN OUTCOME MEASURE(S): Fertilization and pregnancy rates. RESULT(S): In 77% of the patients, spermatozoa could be harvested from the testis by an open testicular biopsy technique and used for intracytoplasmic sperm injection after freezing and thawing of testicular tissue. Histopathologic evaluation revealed a Sertoli cell-only pattern in 21%, maturation arrest in 60%, and hypospermatogenesis in 19% of the patients. In 2. 9% of the patients, carcinoma in situ or a germ cell tumor was detected. In all patients, viable spermatozoa could be visualized after the tissue samples were thawed. One hundred thirty-five intracytoplasmic sperm injection treatment cycles were performed, with a fertilization rate of 45% and a clinical pregnancy rate of 30% per oocyte retrieved. CONCLUSION(S): The use of frozen-thawed testicular tissue allows ovarian stimulation of the female partner to be timed and avoids cancellation of ovum pick-up when spermatozoa cannot be retrieved. PMID- 10688996 TI - Relationship between oxidative stress, semen characteristics, and clinical diagnosis in men undergoing infertility investigation. AB - OBJECTIVE: To determine whether particular semen characteristics in various clinical diagnoses of infertility are associated with high oxidative stress and whether any group of infertile men is more likely to have high seminal oxidative stress. Reactive oxygen species (ROS) play an important role in sperm physiological functions, but elevated levels of ROS or oxidative stress are related to male infertility. DESIGN: Measurement of sperm concentration, motility, morphology, seminal ROS, and total antioxidant capacity (TAC) in patients seeking infertility treatment and controls. SETTING: Male infertility clinic of a tertiary care center. PATIENT(S): One hundred sixty-seven infertile patients and 19 controls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Semen characteristics, seminal ROS, and TAC in samples from patients with various clinical diagnoses and controls. RESULT(S): Fifteen patients (9.0%) were Endtz positive and 152 (91.0%) Endtz negative. Sperm concentration, motility, and morphology were significantly reduced in all groups compared with the controls (P =.02), except in varicocele associated with infection group. Mean (+/-SD) ROS levels in patient groups ranged from 2.2 +/- 0.13 to 3.2 +/- 0.35, significantly higher than controls (1.3 +/- 0.3; P<.005). Patient groups had a significantly lower mean (+/-SD) TAC from 1014.75 +/- 79.22 to 1173.05 +/- 58.07 than controls (1653 +/- 115.28, P<.001), except in the vasectomy reversal group (1532.02 +/- 74.24). Sperm concentration was negatively correlated with ROS both overall and within all groups (P/=8 microg ml(-1) and confirmation by double-disk test, we detected 15 E. coli and 34 K. pneumoniae isolates producing ESBLs. Genotypes of ESBLs determined by PCR with type-specific primers included one TEM-derived and 24 SHV-derived ESBLs, in addition to 24 Toho-1-type ESBLs, one of the major types of ESBLs reported in Japan. Nucleotide sequence analysis of SHV-specific PCR products revealed that SHV-12 was the dominant type of SHV-derived ESBL. In addition, we also identified TEM-26 and SHV-2. This is the first report characterizing TEM- and SHV-derived ESBLs in Japan. PMID- 10689166 TI - The pseudomonas aeruginosa motR gene involved in regulation of bacterial motility. AB - A mini-Tn5-Hg insertion mutant derived from Pseudomonas aeruginosa W51D (W51M1) was isolated in which mini-Tn5 insertion disrupted the motR gene showing that it forms part of the cluster involved in bacterial motility and chemotaxis. Characterization of the W51M1 motility behavior, and also of a PAO1 motR::mini Tn5-Hg mutant, suggests that the product of the motR gene is a negative regulator of bacterial motility which controls the number of flagella per cell. PMID- 10689167 TI - Genomic variations of Mycoplasma capricolum subsp. capripneumoniae detected by amplified fragment length polymorphism (AFLP) analysis. AB - The genetic diversity of Mycoplasma capricolum subsp. capripneumoniae strains based on determination of amplified fragment length polymorphisms (AFLP) is described. AFLP fingerprints of 38 strains derived from different countries in Africa and the Middle East consisted of over 100 bands in the size range of 40 500 bp. The similarity between individual AFLP profiles, calculated by Jaccard's coefficient, ranged from 0.92 to 1.0. On the basis of the polymorphisms detected, the analysed strains can explicitly be grouped into two major clusters, equivalent to two evolutionary lines of the organism found by 16S rDNA analysis. The present data support previous observations regarding genetic homogeneity of M. capricolum subsp. capripneumoniae, and confirm the two evolutionary lines of descent found by analysis of 16S rRNA genes. PMID- 10689168 TI - Energetics of the effect of acetic acid on growth of Saccharomyces cerevisiae. AB - In batch cultures of a respiratory deficient mutant of Saccharomyces cerevisiae the maximum specific growth rate and the yield coefficient decreased, but the specific glucose consumption rate increased, in the presence of acetic acid. The ATP yield decreased from approximately 14 to 4 g biomass (mol ATP)(-1) when the concentration of acetic acid increased from 0 to 170 mM. Intracellular acidification was much weaker than previously reported for non-adapted cells. A linear relation was obtained between the ATP specific production rate and the uptake rate of acetic acid, suggesting that about 1 mol ATP is consumed per mol of acetic acid diffusing into the cells. PMID- 10689169 TI - Changes to water repellence of soil caused by the growth of white-rot fungi: studies using a novel microcosm system. AB - A microcosm system is described which permits assessment of the progressive growth of filamentous fungi through soil. We report on its application to measure the effects of Coriolus versicolor and Phanerochaete chrysosporium upon the sorptivity and water repellence of a mineral soil, measured using a miniature infiltration device. Both fungal species caused moderate sub-critical repellence. Since the pore structure was unaffected, the repellence was probably due to hydrophobic substances of fungal origin. This is the first report of changes in soil repellence caused by the growth of potential xenobiotic bioremediating fungi. The potential consequences are discussed. PMID- 10689170 TI - Clonal turnover of enterohemorrhagic Escherichia coli O157:H7 in experimentally infected cattle. AB - A total of 401 enterohemorrhagic Escherichia coli (EHEC) O157:H7 isolates from two experimentally infected calves were analyzed using molecular biological methods. Genetic differences detected by pulsed-field gel electrophoresis were observed between the inoculated and recovered strains as early as 1 day post inoculation. The loss of the inoculated clone was observed in one calf. Replication and dissemination of the EHEC O157:H7 strains that mutated in cattle may result in the diversification of this organism among cattle populations. PMID- 10689171 TI - Characterisation of extended-spectrum beta-lactamases of the SHV family using a combination of PCR-single strand conformational polymorphism (PCR-SSCP) and PCR restriction fragment length polymorphism (PCR-RFLP). AB - Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) has been developed to extend the identification of SHV beta-lactamases previously characterised by PCR-single strand conformational polymorphism (PCR-SSCP) analysis alone. Eight bacteria, each producing a different SHV beta-lactamase, were used in this study. These bacteria harbour bla(SHV-1), bla(SHV-2a), bla(SHV 3), bla(SHV-4), bla(SHV-5) (two strains), bla(SHV-11) and bla(SHV-12). All isolates were characterised by PCR-SSCP and PCR-RFLP with DdeI and NheI digestion. By a combination of these techniques, the genes encoding these beta lactamases could be differentiated from each other. In addition, the PCR-RFLP technique theoretically can be applied to distinguish the genes encoding SHV-7, SHV-9, SHV-10, SHV-15, SHV-17 and SHV-24 from those encoding other SHV variants. We report a simple PCR-RFLP technique that can be used in epidemiological studies to enable the rapid characterisation of known SHV beta-lactamases in a combination with the previously published PCR-SSCP analysis. PMID- 10689172 TI - Human milk fractions inhibit the adherence of diffusely adherent Escherichia coli (DAEC) and enteroaggregative E. coli (EAEC) to HeLa cells. AB - Binding to a specific receptor is an essential step for most enteropathogens to initiate an intestinal infection. We analyzed the inhibitory effect of human milk and its protein components on adhesion of two diarrheagenic Escherichia coli strains, diffusely adherent E. coli (DAEC) and enteroaggregative E. coli (EAEC), to HeLa cells. Defatted milk, whey proteins, immunoglobulin and non immunoglobulin fractions, in concentrations lower than usually found in whole milk, inhibited both DAEC and EAEC adhesion, indicating that human milk components may contribute to the defense of the infants against enteropathogens. PMID- 10689173 TI - Response of soybean rhizosphere communities to human hygiene water addition as determined by community level physiological profiling (CLPP) and terminal restriction fragment length polymorphism (TRFLP) analysis. AB - In this report, we describe an experiment conducted at Kennedy Space Center in the biomass production chamber (BPC) using soybean plants for purification and processing of human hygiene water. Specifically, we tested whether it was possible to detect changes in the root-associated bacterial assemblage of the plants and ultimately to identify the specific microorganism(s) which differed when plants were exposed to hygiene water and other hydroponic media. Plants were grown in hydroponics media corresponding to four different treatments: control (Hoagland's solution), artificial gray water (Hoagland's+surfactant), filtered gray water collected from human subjects on site, and unfiltered gray water. Differences in rhizosphere microbial populations in all experimental treatments were observed when compared to the control treatment using both community level physiological profiles (BIOLOG) and molecular fingerprinting of 16S rRNA genes by terminal restriction fragment length polymorphism analysis (TRFLP). Furthermore, screening of a clonal library of 16S rRNA genes by TRFLP yielded nearly full length SSU genes associated with the various treatments. Most 16S rRNA genes were affiliated with the Klebsiella, Pseudomonas, Variovorax, Burkholderia, Bordetella and Isosphaera groups. This molecular approach demonstrated the ability to rapidly detect and identify microorganisms unique to experimental treatments and provides a means to fingerprint microbial communities in the biosystems being developed at NASA for optimizing advanced life support operations. PMID- 10689174 TI - Proline accumulation by mutation or disruption of the proline oxidase gene improves resistance to freezing and desiccation stresses in Saccharomyces cerevisiae. AB - We examined the role of intracellular proline under freezing and desiccation stress conditions in Saccharomyces cerevisiae. When cultured in liquid minimal medium, the proline-nonutilizing mutant containing the put1 mutation (proline oxidase-deficient) produced more intracellular proline, and increased the cell survival rate as compared to the wild-type strain after freezing and desiccation. We also constructed two PUT1 gene disruptants. PUT1-disrupted mutants in minimal medium supplemented with external proline at 0.1% accumulated higher proline levels than those of the control strains (17-22-fold). These disruptants also had a 2-5-fold increase in cell viability compared to the control strains after freezing and desiccation stresses. These results indicate that proline has a stress-protective function in yeast. PMID- 10689175 TI - The extracellular hyaluronidase gene (hylA) of Streptococcus pyogenes. AB - Group A streptococci produce an extracellular hyaluronidase (hyaluronate lyase) which may be associated with the spread of the organism during infection. The gene for this hyaluronidase (hylA) encodes an 868 amino acid protein with a molecular size of 99636 Da. Cleavage of the proposed signal peptide results in an extracellular protein of 95941 Da. Comparison with other bacterial hyaluronidases indicates strong similarities to the genes from Streptococcus pneumoniae, Streptococcus agalactiae and Staphylococcus aureus. A region internal to the hylA gene was amplified from all 175 strains of Streptococcus pyogenes tested suggesting a widespread distribution of the gene. PMID- 10689176 TI - Aerobic nitrate respiration in a nitrite-oxidising bioreactor. AB - The ability of heterotrophic bacteria in a nitrite-oxidising bioreactor to respire with nitrate as an electron acceptor was examined. Approximately 70% of 1000 heterotrophic isolates were able to express a nitrate reductase. A detailed survey of 15 isolates showed that five expressed the azide-insensitive nitrate reductase encoded by the napA gene. A two-round PCR amplification of the napA gene using degenerate PCR primers and DNA sequence analysis of these products confirmed the presence of this gene in the positive isolates. Partial 16S rDNA products and napA products were amplified from the biomass in the bioreactor and denaturing gradient gel electrophoresis of these products identified 21 distinct ribotypes and 12 distinct napA sequences. The results show that the ability to respire with nitrate as an electron acceptor under aerobic conditions is widespread among the heterotrophic population of this bioreactor. PMID- 10689177 TI - Transcriptional regulation of the pas gene of enterohemorrhagic Escherichia coli. AB - The Pas protein plays a key role in the pathogenesis of enterohemorrhagic Escherichia coli (EHEC), being required for the secretion of the Esp proteins. Here, the transcriptional regulation of the pas gene was analyzed through the construction of a pas::lacZ translational fusion. When bacteria were grown in Luria Bertani medium or tissue culture medium supplemented with HEPES, a bimodal activation curve was observed. The early phase of induction was not significantly modified by the incubation temperature (either 25 or 37 degrees C), whereas the second phase, which overlaps with the late exponential growth phase, was enhanced at 37 degrees C. The early phase was also stimulated by growth on tissue culture medium and by the addition of Ca(2+), Mn(2+)or Mg(2+) to the M9-glucose minimal medium. Primer extension analysis showed the presence of two major starts of transcription, which were located 58 and 60 bp upstream of the ATG-start codon of the Pas protein, respectively. Although these sites are very close to each other, the transcripts produced during the early induction phase mainly start on the -60 position, whereas the -58 start was activated during the second induction phase. PMID- 10689178 TI - A second tonB gene in Pseudomonas aeruginosa is linked to the exbB and exbD genes. AB - The exbBD genes of Pseudomonas aeruginosa PAO were cloned by complementation of the growth defect of an Escherichia coli exbB tolQ double mutant on iron restricted medium. Nucleotide sequence analysis confirmed that these genes are contiguous and preceded by a second tonB gene in this organism, which we have designated tonB2. lacZ promoter fusions confirmed that expression of the tonB2 exbB-exbD genes is increased under conditions of iron limitation. Deletions within any of these genes, in contrast to deletions in the first tonB gene, tonB1, did not adversely affect growth on iron-restricted medium. On the other hand, tonB1 tonB2 double mutants were more compromised as regards growth in an iron-restricted medium than a tonB1 deletion, indicating that TonB2 could partially replace TonB1 in its role in iron acquisition. TonB1 but not TonB2 deletion strains were also compromised as regards the utilization of hemin or hemoglobin as sole iron sources, indicating that heme transport requires TonB1. PMID- 10689179 TI - Sequence polymorphisms within the pMGA genes and pMGA antigenic variants in Mycoplasma gallisepticum. AB - Antigenic variants of Mycoplasma gallisepticum major surface lipoprotein, pMGA, are encoded by a large gene family. In this study sequence analyses of the PCR amplified pMGA genes showed two types of sequences similar to the pMGA1.2 gene in M. gallisepticum strains. They differed in the sequence encoding a proline-rich region (PRR) at the N-terminus of the pMGA protein. The type A genes had sequences similar to the published pMGA1.2 gene sequence of strain S6, whereas the type B genes lacked the second repetitive segment encoding PTPN sequence within PRR and were similar to the published sequence of PG31 strain. Low in vitro passages of M. gallisepticum strains isolated recently in Slovenia from four avian species showed very different expression patterns of pMGA1.2 and pMGA1.9 genes. Among isogenic populations of S6(B) and IHB1 strains a high frequency of pMGA antigenic variants lacking an epitope for monoclonal antibody (mAb) 71 was found. Strain IHB1 clones, which synthesized pMGA recognized by mAb 71, transcribed pMGA genes whose partial sequence encoded the amino acid sequence (262)TNGDEPRSVS of the mAb 71 epitope. Other IHB1 clones synthesized pMGA variants with different isoelectric points, lacking the epitope for mAb 71, but expressing downstream epitopes for other mAbs. Our study suggests that a molecular basis for pMGA antigenic variation lies in the corresponding changes at the DNA level. PMID- 10689181 TI - Nuclear topography of the c-myc gene in human leukemic cells. AB - The c-myc gene plays an essential role in the regulation of the cell cycle and differentiation. Therefore, changes of the c-myc positioning during differentiation are of great interest. As a model system of cell differentiation, the HL-60 and U-937 human leukemic cell lines were used in our experiments. These cells can be induced to differentiation into granulocytes that represent one of the pathways of blood cell maturation. In this study, changes of the topographic characteristics of the c-myc gene (8q24), centromeric region of chromosome 8 and chromosome 8 domain during differentiation of HL-60 and U-937 cells were detected using fluorescence in-situ hybridisation (FISH). FISH techniques and fluorescence microscopy combined with image acquisition and analysis (high-resolution cytometry) were used in order to detect the topographic features of nuclear chromatin. Increased centre of nucleus-to-gene and gene-to-gene distances of c myc genes, centromeric region of chromosome 8 and chromosome 8 domains were found early after the induction of granulocytic differentiation by dimethyl sulfoxide (DMSO) or retinoic acid (RA); the size of the chromosome 8 domains was rapidly reduced. In differentiated cells, c-myc is located at greater distances from the centromeric regions of chromosome 8. These results support the idea that relocation of the c-myc gene to the nuclear periphery and the condensation of the chromosome 8 domain might be associated with the c-myc gene expression due to common kinetics during granulocytic differentiation. PMID- 10689182 TI - Stable shuttle vectors for Neisseria gonorrhoeae, Haemophilus spp. and other bacteria based on a single origin of replication. AB - An origin of replication (ori) was obtained from a naturally occurring beta lactamase-producing plasmid isolated from Neisseria gonorrhoeae and used to construct shuttle vectors capable of replicating in N. gonorrhoeae, Haemophilus ducreyi, Haemophilus influenzae and Escherichia coli. Using the gonococcal proAB genes, we complemented proline-requiring N. gonorrhoeae F62 and E. coli HB101 in trans. The first demonstration of the expression of the green fluorescent protein (GFP) in either N. gonorrhoeae or H. ducreyi was shown using this vector, indicating that GFP may be a useful tool in the analysis of these organisms. This is the first report of a gonococcal vector based on a broad host range, genetically defined ori, and should facilitate the molecular analysis of gonococcal and Haemophilus genes. PMID- 10689183 TI - PEBP2alphaA/CBFA1 mutations in Japanese cleidocranial dysplasia patients. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant human bone disease whose genetic locus has been located on chromosome 6p21, where the PEBP2alphaA/CBFA1 gene essential for osteogenesis also maps. Previously, several heterozygous mutations in PEBP2alphaA/CBFA1 were found in CCD patients. In this study, we identified six different types of mutations in PEBP2alphaA/CBFA1 in Japanese CCD patients. Four cases were similar to those reported previously: two were nonsense mutations in the Runt domain, one was a hemizygous deletion, and the other was a missense mutation in the Runt domain which abolished the DNA-binding activity of Runx2/PEBP2alphaA/CBFA1. The remaining two mutations were novel: one had a heterozygous gt-to-tt mutation at the splice donor site (gt) between the exon3 intron junction, which resulted in abnormal exon3 skipping, and the other had a mutation in exon7, which led to the introduction of a translational stop codon in the middle of the transactivation domain. Thus, defects in either the DNA-binding domain or transactivation domain of Runx2/PEBP2alphaA/CBFA1 can cause CCD. The results not only provide a strong genetic evidence that mutations involving in PEBP2alphaA/CBFA1 contribute to CCD, but also provide a useful tool to study how Runx2/PEBP2alphaA/CBFA1 plays its pivotal role during osteoblastic differentiation. PMID- 10689184 TI - MSY2: a slowly evolving minisatellite on the human Y chromosome which provides a useful polymorphic marker in Chinese populations. AB - We present the second human Y-specific minisatellite, MSY2 (DYS440). It consists of three or four copies of a 99-110bp repeat unit and is located about 1kb upstream of the DBY gene. The most common allele contains four units, but a three unit allele has arisen on at least four occasions; in chimpanzees and orangutans, MSY2 contains only two units. It is therefore evolving slowly and provides a particularly useful polymorphic marker for Chinese populations. PMID- 10689185 TI - Structure of the murine Pit1 phosphate transporter/retrovirus receptor gene and functional characterization of its promoter region. AB - The Pit1 phosphate transporter (formerly also called Glvr-1) probably plays an important role in regulated phosphate handling in bone-forming cells. In this study, we describe the structure of the mouse Pit1 gene, as well as some functional characteristics of its promoter region in murine bone cells. Screening of a genomic library led to the isolation of two overlapping lambda clones containing 7kb of 5' flanking region, as well as the 10 exons of the mouse Pit1 gene corresponding to the published cDNA. The translation start site is located within exon I and the stop codon within exon X. The overall structure of the mouse gene is very similar to that of its human homolog, except for the presence of an additional 5' untranslated exon in human. The structure of the 5' untranslated region of the mouse gene was thus further investigated using rapid amplification of cDNA ends in murine ATDC5, MC3T3-E1 and Swiss 3T3 cells. The results indicate that, compared to the published cDNA, the mouse Pit1 gene contains in fact one additional 5' exon, which we named exon IA. Reporter gene assays demonstrate the presence of a functional TATA box containing promoter upstream of exon IA. This description of the murine Pit1 gene and of its promoter region paves the way to more detailed analyses concerning the regulation of Pit1 transcription in mouse cells. Furthermore, a comparison of mouse and human promoters will hopefully allow a better understanding of general mechanisms regulating Pit1 expression in different species. PMID- 10689186 TI - Mammalian genomes contain active recombinase recognition sites. AB - Recombinases derived from microorganisms mediate efficient site-specific recombination. For example, the Cre recombinase from bacteriophage P1 efficiently carries out recombination at its loxP target sites. While this enzyme can function in mammalian cells, the 34bp loxP site is expected to be absent from mammalian genomes. We have discovered that sequences from the human and mouse genomes surprisingly divergent from loxP can support Cre-mediated recombination at up to 100% of the efficiency of the native loxP site in bacterial assays. Transient assays in human cells demonstrate that such pseudo-lox sites also support Cre-mediated integration and excision in the human cell environment. Pseudo sites for Cre and other recombinases may be useful for site-specific insertion of exogenous genes into mammalian genomes during gene therapy and other genetic engineering processes. PMID- 10689187 TI - Gene structure and chromosome mapping of mouse transcription elongation factor S II (Tcea1). AB - We report the organization and chromosome localization of the mouse transcription elongation factor S-II gene (Tcea1). This gene was found to be a single copy gene consisting of 10 exons spanning approximately 30kb. Its organization was the same as those of the mouse testis-specific S-II gene (Tcea2) and Xenopus general S-II gene (xTFIIS.oA), but different from that of the human S-II gene family. We also identified a processed pseudogene (Tcea1-ps1) with a sequence highly homologous to those of S-II cDNAs but containing a translation termination codon within its open reading frame. Linkage analysis showed that Tcea1 and Tcea1-ps1 are mapped on mouse chromosomes 1 and 15, respectively. Relationships between Tcea1 and S-II cDNAs isolated so far are discussed. PMID- 10689188 TI - Molecular structure and tissue-specific expression of the mouse pancreatic phospholipase A(2) gene. AB - Pancreatic phospholipase A(2) (PLA(2)) is involved with the hydrolysis of phospholipids into lysophospholipids and unesterified fatty acids. The enzyme has been postulated to play a key role in lipid absorption by intestinal absorptive cells as well as in the regulation of secretin release from intestinal endocrine cells. This manuscript reports the genomic organization and the primary sequence of the mouse PLA(2). The results showed that the mouse PLA(2) gene contains four exons interspersed by three introns, spans over 8kb in length, and is considerably larger than the human PLA(2) gene. The mouse PLA(2) protein contains 146 amino acid residues, including the signal peptide. The mouse protein is highly homologous to the rat, dog, and human enzyme, but is two residues shorter than the human protein. Mouse PLA(2) message is synthesized predominantly in the pancreas, but the lung also contains low levels of PLA(2) mRNA. PMID- 10689189 TI - Matrix attachment region regulates basal beta-lactoglobulin transgene expression. AB - Nuclear matrix attachment regions (MAR) have been implicated in the regulation of gene expression. We have identified a region within the proximal 3'-flanking sequences of the ovine beta-lactoglobulin (betalg) gene that interacts with the nuclear matrix in vitro. No equivalent region was detected in the 5' flanking region. We have investigated the role of this element in regulating betalg expression in vitro and in vivo. Removal of the MAR did not affect the frequency of betalg transgene expression at the mRNA level, but betalg transgenes that lacked the MAR were expressed at a lower level than wild-type betalg transgenes. In neither in-vitro HC11 transfection experiments nor transgenic mice was hormonal induction of betalg expression significantly affected by MAR removal. Nuclear run-on analysis demonstrated that the impaired basal expression of betalg transgene loci lacking the MAR was due to a reduced transcription rate. Thus, the single MAR enhances the basal transcriptional potential of the betalg gene. PMID- 10689190 TI - Cloning of a calcitonin gene-related peptide receptor and a novel calcitonin receptor-like receptor from the gill of flounder, Paralichthys olivaceus. AB - For the first time in non-mammalian vertebrates, cDNA encoding CGRPR was isolated from the gill cDNA library of flounder. The nucleotide sequence consists of a 237bp 5'-UTR, a 1398bp coding sequence for a 465-amino-acid protein, and a 981bp 3'-UTR. The predicted amino-acid sequence has a high degree of identity to hCGRPR (72.3%) and rCGRPR (71.6%) and, to a lesser degree, to hCTR (55.6%) and rCTR (59.3%). In addition, a different type of receptor cDNA was also obtained from the gill cDNA. The nucleotide sequence contains an open-reading frame of 1380bp to produce a 459-amino-acid protein. The open-reading frame of this receptor shows the same degree of identity to mammalian CTR (60.2% to hCTR; 62.3% to rCTR) and CGRPR (64.6% to hCGRPR; 64.4% to rCGRPR). However, the predicted amino-acid sequence was more homologous to hCGRPR (60.2%) and rCGRPR (61.3%) than to hCTR (48.8%) and rCTR (49.5%). The identity of this receptor to fCGRPR is 66.6% at the nucleotide level and 64.2% at the amino-acid level, indicating that the receptor is not likely to be an isoform of CGRPR. The receptor, but not fCGRPR, is expressed in bones, suggesting the possibility that this receptor corresponds to the flounder CTR. PMID- 10689191 TI - The PAUSE software for analysis of translational control over protein targeting: application to E. nidulans membrane proteins. AB - The PAUSE software has been developed as a new tool to study translational control over protein targeting. This makes it possible to correlate the position of clusters of rare codons in a gene, predicted to cause a translational pause, with the position of hydrophobic stretches in the encoded protein, predicted to span a membrane or to act as a cleavable signal for targeting to the secretory pathway. Furthermore, this software gathers these correlations over whole sets of genes. The PAUSE software is described here, and its use is illustrated on a set of membrane proteins from the fungus Emericella nidulans. Preferential distances of about 45 codons and of about 70 codons between putative transmembrane domains and predicted translational pauses were observed. Given that approximately 30 residues are required to span the large ribosomal subunit, the predicted pauses would therefore occur when the hydrophobic domain starts protruding from the ribosome ('+45 pause'), or fully protrudes as a hairpin ('+70 pause'). Thus, these specific pauses might reflect a translational control over membrane protein targeting or early recognition ('+45 pause'), and over insertion or folding ('+70 pause'). PMID- 10689192 TI - Description and characterization of IS994, a putative IS3 family insertion sequence from the salmon pathogen, Renibacterium salmoninarum. AB - Renibacterium salmoninarum, a slowly growing, Gram-positive bacterium, is responsible for bacterial kidney disease in salmonid fishes world-wide. To date, no mobile genetic elements have been reported for this pathogen. Here, we describe the first insertion sequence (IS) identified from R. salmoninarum. This element, IS994, has a significant predicted amino acid sequence homology (64.8 and 71.9%) to the two open reading frames encoding the transposase of IS6110 of Mycobacterium tuberculosis. Protein parsimony and protein distance matrix analyses show that IS994 is a member of group IS51 of the IS3 family. From a conservative estimate, there are at least 17 chromosomal insertions of IS994 or closely related elements. Sequence analysis of seven of these loci reveals single nucleotide polymorphisms throughout the element (including the terminal inverted repeats), a 15bp insertion in three of the seven loci, and an absence of flanking direct repeats or conserved insertion site. Restriction fragment length polymorphism analysis of XbaI-digested chromosomal DNA shows variations among European and North American isolates, indicating that IS994 may be a useful molecular marker for epizootiological studies. PMID- 10689193 TI - Isolation and functional characterization of a temperature-sensitive mutant of the yeast Saccharomyces cerevisiae in translation initiation factor eIF5: an eIF5 dependent cell-free translation system. AB - Eukaryotic translation initiation factor 5 (eIF5) interacts with the 40S ribosomal initiation complex (40S.eIF3.AUG.Met-tRNA(f).eIF2.GTP) to promote the hydrolysis of bound GTP. In Saccharomyces cerevisiae, eIF5, a protein of 45346Da, is encoded by a single-copy essential gene, TIF5. In this paper, we have isolated a temperature-sensitive S. cerevisiae strain, TMY5-1, by replacing the wild-type chromosomal copy of TIF5 with one mutagenized in vitro. The mutant yeast cells rapidly cease protein synthesis when grown under non-permissive conditions, lose polyribosomes and accumulate free 80S ribosomes. Further characterization of mutant eIF5 showed that the mutant protein, expressed in Escherichia coli, is defective both in its interaction with eIF2 as well as in mediating the hydrolysis of GTP bound to the 40S initiation complex and consequently in the formation of the 80S initiation complex. Additionally, the availability of a yeast strain containing temperature-sensitive mutation in the eIF5 gene allowed us to construct a cell-free translation system that was dependent on exogenously added eIF5 for translation of mRNAs in vitro. PMID- 10689194 TI - Hyperactivation of the Drosophila Hop jak kinase causes the preferential overexpression of eIF1A transcripts in larval blood cells. AB - Jak kinase-Stat protein pathways play a critical role in the response of blood cells to a range of cytokines and growth factors. We are using the fruit fly, Drosophila melanogaster, as a model system to elucidate additional components of Jak-Stat pathways, and to determine how abnormalities in this pathway lead to hematopoietic leukemia-like defects. To identify downstream targets, we conducted a molecular screen for genes whose transcripts are overexpressed in response to activation of the Drosophila Hop Jak kinase. We identified a Drosophila homolog of eIF1A, a eukaryotic initiation factor found in humans and other eukaryotes. D eIF1A is highly overexpressed in the hemocytes and lymph glands of third instar larvae carrying the dominant, gain-of-function mutation hop(Tum-l). A quantitative comparison of poly(A)(+) RNA levels between D-eIF1A and other known Drosophila translation initiation factors indicates that D-eIF1A transcripts preferentially overaccumulate in response to the hyperactive Hop pathway. Our results support the model that D-eIF1A is one of the target genes through which the Drosophila Jak kinase pathway regulates hemocyte development. PMID- 10689195 TI - Isolation and characterization of copia-type retrotransposons in Arabidopsis thaliana. AB - We isolated two copia-type retrotransposons from Arabidopsis thaliana. We named these elements AtRE1 (Arabidopsis thaliana Retro Element 1) and AtRE2. Nucleotide sequence analysis revealed that both elements have long terminal repeats (LTRs), and that their internal sequences include one large open reading frame that could encode Gag protein, protease, integrase, reverse transcriptase, and RNaseH. The deduced amino acids sequences contain several domains that are conserved among a large family of retrotransposons. The primer binding site for first-strand DNA synthesis and the polypurine tract for second-strand DNA synthesis existed at corresponding positions. A 5bp target site duplication (TSD) sequence was also found in the flanking region of LTRs. Southern hybridization and sequence determination of the flanking region demonstrated that AtREs exist at different loci in the two A. thaliana ecotypes Columbia and Landsberg erecta. Moreover, AtRE2 exists at two loci in Landsberg erecta, in contrast to the existence of only one copy in Columbia. These findings suggest that AtREs were recently transfected via some mediators or that AtREs were transposed after differentiation of the two ecotypes. One cDNA clone derived from the transcripts of AtRE1 was isolated, and the nucleotide sequence showed that this RNA was transcribed in the antisense direction. RT-PCR analysis revealed that AtRE1 was transcribed in both directions. This result suggests that the antisense RNA controls the expression of AtRE1 at the post-transcriptional level. PMID- 10689196 TI - Molecular organization of the mouse gastrin-releasing peptide receptor gene and its promoter. AB - The murine gastrin-releasing peptide receptor (mGRP-R) is a member of the G protein-coupled receptor family and mediates important physiological actions of its specific ligand, the gastrointestinal hormone/neurotransmitter GRP, including mitogenic properties in the mouse Swiss 3T3 fibroblasts. Glucocorticoids and increases in intracellular cAMP are reported to alter GRP-R gene transcription, but the molecular basis for these effects is unknown. To begin to identify possible gene regulatory mechanisms that are responsible for modifying mGRP-R expression, we determined its structure and investigated its basal promoter activity. We isolated and characterized genomic bacteriophage P1 clones encoding the mouse gastrin-releasing peptide receptor (mGRP-R). By DNA sequencing and Southern blot analyses, we determined the protein coding region to be contained in three exons interrupted by two introns 20 and 2kb in length. The open reading frame of the putative GRP-R gene encodes for a 384-amino-acid protein which demonstrates 48% identity with the mouse BRS-3 protein and 53% identity with the mouse NMB-R protein. The mGRP-R gene locus extends over 29kb and was mapped to the X-chromosome (DXMit20) utilizing a minisatellite polymorphism in the 5' UTR and by fluorescent in-situ hybridization (FISH). In Swiss 3T3 cells, which natively express mGRP-R, two gene-specific mRNA species of 3 and 7kb can be detected by Northern blot analysis. With RNase protection assays, and independently with inverse PCR of 5' RACE clones, common mRNA initiation sites were identified clustered between 21 and 61bp downstream of a TTTAAA motif, which is located 450bp upstream of the ATG translation start site. However, different polyadenylation sites are utilized. A 2kb genomic DNA fragment extending from 2147 to 141 bases 5' to the ATG translation start was cloned into a luciferase reporter plasmid and shown to contain promoter activity in Swiss 3T3 and COS-7 cells. Progressive promoter truncations and mutations of a cyclic AMP response element (CRE) located 83bp upstream of the TTTAAA motif demonstrate that transcriptional mGRP-R activation in Swiss 3T3 cells only occurs when both the TTTAAA motif and the intact CRE site are retained. With the availability of the full structure of the mGRP-R gene and the minimal promoter sequences reported in this study, it will be possible in future studies to investigate the molecular basis for transcriptional regulation of the mGRP-R gene by glucocorticoids, cAMP and other factors. PMID- 10689197 TI - The stachydrine catabolism region in Sinorhizobium meliloti encodes a multi enzyme complex similar to the xenobiotic degrading systems in other bacteria. AB - Stachydrine (proline betaine) can be used by Sinorhizobium meliloti as a source of carbon and nitrogen. Catabolism depends on an initial N-demethylation, after which the resultant N-methyl proline enters general metabolism. Deletion and insertion mutagenesis demonstrated that the information necessary for catabolism is carried on the symbiotic plasmid (pSym) distal to nodD2 and the nod-nif cluster. Sequencing of an 8.5kb fragment spanning this region revealed four open reading frames with functional homology to known proteins, including a putative monooxygenase and a putative NADPH-FMN-reductase, which were shown by insertional and frame-shift mutagenesis to be necessary for stachydrine catabolism. Other open reading frames, encoding a putative flavoprotein and a repressor, were judged not to be required for stachydrine catabolism, since they were not included in a fragment capable of complementing a deletion of the entire stc region. Sequence and mutagenesis data suggest that stachydrine is demethylated by an iron-sulfur monooxygenase of the Rieske type with a requirement for a specific reductase. The stc catabolic cluster, therefore, resembles xenobiotic degradation in other bacteria and recalls rhizopine catabolism in S. meliloti. Stachydrine appears to have multiple roles in osmoprotection, nutrition and nodulation. Genes involved in stachydrine catabolism are also necessary for carnitine degradation; thus, they could be important in the catabolism of a variety of root exudates and mediate other relationships. PMID- 10689198 TI - Valproic acid-induced alterations in growth and neurotrophic factor gene expression in murine embryos [corrected]. AB - Although the teratogenicity of valproic acid (VPA) has been well established, the mechanism(s) by which this anticonvulsant drug induces malformations remains controversial. Using the combined molecular techniques of in situ-transcription (IST) and antisense RNA (aRNA) amplification we analyzed VPA-induced alterations in the gene expression for 10 genes within the neural tubes of embryos from two murine strains that have been shown to differ in their susceptibility to VPA induce neural tube defects (NTD). Pregnant dams from both SWV (susceptible) and LM/Bc (resistant) strains were either treated with saline (control) or VPA (600 mg/kg) on gestational day (GD) 8:12 (day:hour). Neural tubes were isolated from control or VPA exposed embryos at three gestational time points, which represented the beginning (GD 8:18), middle (GD 9:00), and end (GD 9:12) of neural tube closure (NTC) in both of these murine strains. Using univariant statistics we demonstrated that in LM/Bc embryos with NTDs, the expression of bdnf, ngf, and trk, ngf-R were significantly elevated at all three time points, and the cytokine, cntf was significantly decreased at GD 9:00. In contrast, the major gene alterations observed in SWV embryos were a significant increase in tfgalpha and tgfbeta1-3 at GD 9:00. In an effort to better define the more intricate interactions between VPA exposure and the expression of these genes, we analyzed our data using Principal Component Analysis. The results from this analysis demonstrated that embryos from these two stains behaved differently, not only in response to a VPA exposure, but also under control conditions, which may explain the multifactorial nature of NTDs in these mice. PMID- 10689199 TI - Effects of dibutyl phthalate on reproductive function in pregnant and pseudopregnant rats. AB - In our previous studies, dibutyl phthalate (DBP) was found to be embryolethal and teratogenic in rats. In this study, the effects of DBP on reproductive function were investigated on pregnant and pseudopregnant rats. Rats were given DBP by gastric intubation at 0, 250, 500, 750, 1000, 1250 or 1500 mg/kg on Days 0 to 8 of pregnancy and the pregnancy outcome was determined on Day 20 of pregnancy. The same doses of DBP were given to pseudopregnant rats, with an induced decidual cell response, on Days 0 to 8 of pseudopregnancy, and the uterine weight on Day 9 served as an index of the uterine decidualization. DBP caused significant increases in the incidences of preimplantation loss in females successfully mated at 1250 and 1500 mg/kg and of postimplantation loss in females having implantations at 750 mg/kg and above. The uterine decidualization in pseudopregnant rats was significantly decreased at 750 mg/kg and above. These findings suggest that early embryonic loss due to DBP may be mediated, at least in part, via the suppression of uterine decidualization, an impairment of uterine function. PMID- 10689200 TI - Two-generation reproduction study in rats given di-isononyl phthalate in the diet. AB - The potential reproductive toxicity of di-isononyl phthalate (DINP: CAS RN 68515 48-0) was assessed in one- and two-generation reproductive toxicity studies. Groups of 30 male and female CRL : CD(SD)BR rats were given DINP via dietary administration at levels of either 0.0, 0.5, 1, or 1.5% (one-generation study) or 0.0, 0.2, 0. 4, or 0.8% (two-generation study). There were no changes in any of the classic reproductive parameters, i.e. mating, male or female fertility, fecundity, gestational index, or length of gestation in either study. The overall NOAELs for these effects were the highest Dietary Level (%)s tested, approximately 500 mg/kg/day in the two-generation study and 1000 mg/kg/day in the one-generation study. There were no testicular effects in parental animals exposed as juveniles and young adults at 960 mg/kg/day in the one-generation study. In the two-generation study, there were no testicular effects in either the P(1) males, exposed as juveniles and young adults or the P(2) (F(1)) offspring exposed in utero, through lactation, and continuously to terminal sacrifice. The NOAEL was 470 mg/kg/day. Offspring survival was reduced at the 1.5% level ( approximately 1100 mg/kg/day) but unaffected at the 1% level ( approximately 760 mg/kg/day). There were decreased offspring body weights both at postnatal day (PND) 0 and during lactation; however, the PND 0 effects were only clearly related to treatment at the 1.5% level. Weights of offspring during lactation were significantly reduced but within the historical control range at Dietary Level (%)s below 1%. As there was rapid recovery at the lower levels, even though treatment continued, the toxicologic significance is unclear. Adult survival was unaffected at any level in either study, but weight gain was significantly reduced at the 1% level ( approximately 600 mg/kg/day). Liver and kidney weights were elevated at Dietary Level (%)s above approximately 110 mg/kg/day, consistent with evidence from other studies of peroxisomal proliferation at these levels. This study showed that DINP treatment does not affect fertility or male reproductive development at doses of up to approximately 1000 mg/kg/day. PMID- 10689201 TI - The reversible effects of raloxifene on luteinizing hormone levels and ovarian morphology in mice. AB - Raloxifene is a selective estrogen receptor modulator that has estrogen agonist effects on bone and serum lipids and estrogen antagonist effects on breast and uterine tissues. This study assessed the effects of raloxifene hydrochloride (HCl) treatment on circulating luteinizing hormone (LH) levels and ovarian morphology in sexually mature, 15-week-old, female CD-1 mice. Mice were maintained on diets providing average daily doses of 0 or 233 mg/kg raloxifene for 2 weeks (Study 1) or 0, 7.9, or 236 mg/kg raloxifene for 4 weeks (Study 2). At the end of the treatment period, blood samples were collected every 2 hours for 24 h in Study 1 (5 mice per group) and at 10:00 a.m. and 10:00 p.m. in Study 2 (8 mice per group). Serum LH levels were measured by radioimmunoassay. Ovarian histomorphology was evaluated in the 10 mice per group (Study 1) and the 8 mice per group (Study 2). For the reversibility phase (Study 2), mice were fed untreated diets for 3 weeks; serum LH levels and ovarian histomorphology were then assessed. Raloxifene treatment at 233 mg/kg/day for 2 weeks (Study 1) significantly elevated circulating LH levels by 4- to 7-fold compared with control. Raloxifene-treated mice had elevated LH levels sustained over the 24-h sampling period and did not exhibit the preovulatory LH surge evident in some control mice at the 4:00 p.m., 6:00 p.m., and 8:00 p. m. time points. Mice treated with 236 mg/day raloxifene for 4 weeks (Study 2) had elevated LH levels (4.4-fold compared to control), whereas mice exposed to 7.9 mg/kg/day raloxifene had a slight, nonsignificant increase in LH (2-fold compared to control). In both dose groups, LH levels were indistinguishable from controls 3 weeks after raloxifene treatment was discontinued. The ovaries in six of the eight mice treated with 7.9 mg/kg/day raloxifene had dilated and/or anovulatory follicles. One mouse in this group had a single hemorrhagic follicle; however, corpora lutea distribution was normal, indicating that ovulation was occurring. Raloxifene treated mice in Study 1 and mice treated with a comparable raloxifene dose (236 mg/day) in Study 2 had histomorphological changes in the ovary indicative of arrested follicular maturation, including anovulatory hemorrhagic follicles, some developing follicles, and very few corpora lutea. At the end of the reversibility phase, hemorrhagic follicles were no longer evident and follicular maturation and corpora lutea distribution were normal. Raloxifene treatment in mice produces a dose-dependent, sustained elevation in serum LH levels and is associated with changes in ovarian follicular morphology. These changes are reversible upon discontinuation of raloxifene treatment. PMID- 10689202 TI - In vitro fertilization after in vivo treatment of rats with three reproductive toxicants. AB - One objective of these experiments was to establish a sensitive assay to evaluate fertilizing potential of rat gametes in vitro. A second objective was to evaluate this in vitro fertilization (IVF) assay as a method to detect in vivo effects of reproductive toxicants on male and female gametes using three known reproductive toxicants as model systems. The IVF assay with zona-free oocytes was more precise than the assay with cumulus-intact oocytes in these studies (coefficients of variation of 8.7 and 14.4%, respectively). Sperm fertilizing potential for zona free oocytes was reduced by treatment of rats with m-dinitrobenzene (10-10 000 microg/kg) and ethylene glycol monomethyl ether (50-100 mg/kg) that had no effect on sperm motility. Molinate (60 mg/kg for 5 days) reduced sperm fertilizing potential concurrently with reductions in sperm motility. Neither molinate (60 mg/kg for 5 days) nor dinitrobenzene (0.002% in the drinking water for 14 days) administered to females seemed to affect the fertilizability of their oocytes. Ethylene glycol monomethyl ether treatment (0.15-0.25% in the drinking water for 14 days) reduced the number of ovulated oocytes. IVF is a means to evaluate toxicant effects on female gametes and demonstrates sperm's ability to interact with the oocyte plasma membrane. PMID- 10689203 TI - Rat epididymal sperm motion changes induced by ethylene glycol monoethyl ether, sulfasalazine, and 2,5-hexandione. AB - Epididymal sperm was examined using the Hamilton-Thorne Sperm analyzer (HTM-IVOS, version 10.6) in male rats treated with known male reproductive toxicants that act by different mechanisms to detect effects on sperm motion. Three agents known to produce changes in sperm motion at high exposure levels were administered at lower levels. Ethylene glycol monoethyl ether (EGEE), sulfasalazine (SASP), and 2,5-hexandione (2,5-HD) were administered by oral gavage to adult male Sprague Dawley rats at 250 or 500 mg/kg/day, at 300 or 600 mg/kg/day, or at 100 or 250 mg/kg/day, respectively. The males were treated with EGEE, SASP, and 2,5-HD for 35, 28, and 28 days, respectively. The males treated with EGEE and SASP were mated with untreated females to assess male fertility. All males were examined for body weight, testicular and epididymal weight, epididymal sperm count, and sperm motion. The sperm motion parameters included percentage of motile sperm, percentage of progressively motile sperm (progressive motility), curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), amplitude of lateral head displacement (ALH), beat cross frequency (BCF), linearity (LIN), and straightness (STR). For the male rats treated with SASP, no treatment-related effects on percentages of motile sperm or sperm count were observed despite impaired male fertility. However, abnormal motion of epididymal sperm from the SASP treated males was detected by a significant reduction in mean progressive motility, VAP, and ALH, and an increase in BCF and STR. For the males treated with 2,5-HD for 4 weeks, most parameters generated by the HTM-IVOS indicated decreased sperm motion despite no remarkable changes in testicular weight, epididymal weight, or sperm count. In the EGEE-treated males at 250 mg/kg/day for 5 weeks, abnormal motion of epididymal sperm was detected by decreased progressive motility and increased BCF, although there were no treatment-related effects on testicular weight or male fertility. Progressive motility was decreased in all treated groups and the difference from the control value was of the greatest magnitude among the sperm motion parameters generated by the HTM-IVOS. Velocity parameters (VAP, VSL, VCL) responded sensitively to abnormal sperm motion in the SASP and 2,5-HD studies. In spite of decreased sperm motion, BCF values were significantly increased in all treated groups except the 7-week EGEE high-dose group, where there were no motile sperm to evaluate. ALH was significantly decreased in the treated groups in which remarkable effects on sperm motion were noted. There were no significant changes in ALH at the low-dose of EGEE at which only mild effects on sperm motion were observed. STR was increased for epididymal sperm from the males treated with SASP when compared with the controls. For the males treated with EGEE and 2,5-HD, however, STR was decreased when compared with the controls. There were no significant differences in LIN in any of the groups treated with SASP, in which remarkably reduced sperm motion was detected by the other parameters. In conclusion, among the parameters generated by the HTM-IVOS, progressive motility was significantly decreased in all treated groups and the most valuable for detecting slight changes in sperm motion induced by these three different target toxicants. Further investigation with a larger set of compounds is needed to evaluate which IVOS parameters are the most sensitive in detecting motion changes. PMID- 10689204 TI - Histochemical tracing of bismuth in testis from rats exposed intraperitoneally to bismuth subnitrate. AB - The histochemical silver amplification technique autometallography (AMG), was used to trace bismuth in the testis of Wistar rats injected intraperitoneally with bismuth subnitrate. In the seminiferous tubules, bismuth was located in lysosomes of Sertoli cells closely associated with heads of spermatids in the late stages of the spermatogenesis, i.e. shortly before the release of Step 19 spermatids in Stage XIII. No bismuth-specific AMG silver grains were detected in the spermatogenic cell line. However, tails of free sperm cells located in the tubular lumen showed autometallographic grains in close contact to the nine outer microtubule doublets in the axonema. Leydig cells concentrated huge amounts of AMG-bismuth in their lysosomes. Furthermore, parallel exposure to selenium significantly increased the amount of histochemically traceable bismuth in the rat testis. PMID- 10689205 TI - Dr. Brent and scientific debate. PMID- 10689206 TI - Safe drinking water: An attainable goal, key to health and development, appears farther away. PMID- 10689207 TI - Limited effectiveness of home drinking water purification efforts in Karachi, Pakistan. AB - OBJECTIVE: In many developing-country urban areas, municipally supplied water is not microbiologically safe. This study evaluated drinking water quality and effect of home water purification efforts in Karachi, Pakistan. METHODS: Members of 300 households, including 100 households who used the Aga Khan University Hospital Laboratory and 200 of their neighbors were interviewed. In 293 consenting households, structured observations were performed and drinking water was analyzed for the presence of coliforms, using the multiple tube fermentation technique. RESULTS: Although 193 of the 293 households (66%) reported using some method to purify their drinking water, including 169 (58%) who boiled their water, only 48 (16%) of the drinking water samples were free of coliforms. Although a combination of boiling and filtering was the most effective method of purification, only 38% of samples that had been boiled and filtered were free of coliforms. CONCLUSIONS: Further refinements and evaluations of home-based efforts to purify and store water are needed. PMID- 10689208 TI - Epidemic cholera in Guinea-Bissau: the challenge of preventing deaths in rural West Africa. AB - OBJECTIVES: An epidemiologic investigation was conducted to identify factors associated with cholera mortality in a rural African setting and interventions likely to prevent deaths in future epidemics. METHODS: The authors reviewed surveillance data from rural Biombo, Guinea-Bissau, interviewed family members of persons who died of cholera, and conducted a case-control study in the catchment area of a health center with a high case:fatality ratio (CFR). RESULTS: Forty three deaths occurred among the 1169 persons who reported to health centers with cholera during the epidemic (CFR = 3.7%). Delayed rehydration and over-hydration probably contributed to 10 of these deaths. An additional 19 cholera deaths occurred outside health centers. In the case-control study, persons with cholera who died were 5.4 times (95% CI = 1.0-53.4) more likely to be in poor health or intoxicated at illness onset than persons with cholera who survived. Fatal cases were 6.0 times (95% CI = 1.1-60.8) more likely to not attend the health center than survivors. CONCLUSIONS: The low overall CFR in Biombo, compared to CFRs reported during other epidemics in sub-Saharan Africa, suggests that medical care provided at rudimentary rural health centers prevented numerous deaths. Additional deaths may be prevented by strengthening the infrastructure of health services in the rural areas and by enhanced public education regarding the need for persons with cholera to promptly seek medical care. PMID- 10689209 TI - Introducing a novel model to estimate national and global measles disease burden. AB - OBJECTIVES: In discussions of expanded measles control, elimination, and possible eradication, better estimates of disease burden are increasingly important to target vaccination control measures. Because global surveillance for measles is inadequate, a model to quantify country-specific estimates of measles disease burden was formulated to help policy-makers consider control options. METHODS: Country-specific demographics, developmental status, historic vaccine coverage rates, and age-specific vaccine efficacy and attack rates were used to determine the number of measles cases and deaths for 5-year periods. RESULTS: The model estimates an annual global incidence of 32 million measles-susceptible persons ( approximately 25% of the global birth cohort), resulting in 28 million cases and 691 thousand deaths. Eighty-four percent (578,000) of the global deaths occur in the World Health Organization African and Southeast Asian regions. Twenty countries account for 82% of deaths attributable to measles. In nine countries, over 2% of the birth-cohort are estimated to die from measles. CONCLUSIONS: This methodology quantifies country- and age-specific measles disease burden and establishes regional and global disease patterns, allowing aggregations by income groups and regions, which aids policy formulation. The data may be continuously updated, based on dynamic changes in vaccine coverage rates and the incorporation of national vaccination campaigns. PMID- 10689210 TI - Ceftazidime-resistant Klebsiella pneumoniae bloodstream infection in children with febrile neutropenia. AB - OBJECTIVES: To evaluate prevalence of ceftazidime-resistant Klebsiella pneumoniae (CRKP) in the pediatric oncology unit of University Hospital, Kuala, Lumpur, and to identify differences between febrile neutropenic pediatric patients with CRKP and ceftazidime-sensitive K. pneumoniae (CSKP) bacteremia. MATERIALS AND METHODS: Febrile neutropenic patients treated between January 1996 and December 1997 at the pediatric oncology unit of University Hospital, Kuala Lumpur, were prospectively studied. Empirical antibiotic therapy consisted of ceftazidime and amikacin. Those who developed K. pneumoniae bacteremia were identified, and clinical features analyzed. Ceftazidime-resistance was documented via disk diffusion testing. Production of extended-spectrum beta-lactamase (ESBL) was inferred on the basis of synergy between ceftazidime and amoxicillin-clavulanic acid. The different features between the two groups and variables associated with the development of CRKP bacteremia were analyzed using chi-square and t-tests and calculation of odds ratios. A multivariate analysis was used to identify independent factors for CRKP development. RESULTS: Ceftazidime-resistance was seen in 51.6% of all K. pneumoniae isolates, and all these isolates were inferred to be ESBL producers. All isolates were sensitive to imipenem. Susceptibility to gentamicin was 90.5%. The mean continuous hospital stay prior to the detection of bacteremia was 13.7 days overall, but significantly longer in the CRKP group (21.9 d) compared to the CSKP group (4.3 d) (P = 0.003). Children with CRKP were more likely to have received antibiotics in the 2 weeks prior to detection of bacteremia (87.5% of cases) than the CSKP group (20.0% of cases) (P = 0.0008). Sepsis-related mortality was higher in those with CRKP (50.0%) than in the CSKP group (13.3%) (P = 0.02). Patients who did not receive CRKP-directed antibiotics within 48 hours of admission were more likely to have a fatal outcome than those who did (P = 0.009). Logistic regression analysis identified use of third generation cephalosporins 2 weeks prior to presentation and a hospital stay of 2 weeks or more as independent risk factors for development of CRKP. CONCLUSIONS: More than half of total K. pneumoniae isolated from blood cultures in the unit were ceftazidime-resistant. Children with febrile neutropenia with prolonged hospital stay and recent prior antibiotic exposure are at high risk of developing CRKP bacteremia. Mortality was significantly higher in this group. Early commencement of appropriate antibiotics (e.g., imipenem with or without gentamicin), according to susceptibility study results, may be beneficial in such circumstances. PMID- 10689211 TI - International study comparing cefdinir and cefuroxime axetil in the treatment of patients with acute exacerbation of chronic bronchitis. AB - OBJECTIVES: To assess the efficacy and tolerability of three antibiotic regimens in patients with acute exacerbation of chronic bronchitis. METHODS: In this double-blind, randomized, multicentered, parallel-group study, patients received once-daily cefdinir 600 mg, twice-daily cefdinir 300 mg, or twice-daily cefuroxime axetil 250 mg for 10 days. Primary efficacy measures were microbiologic eradication rate, by pathogen and by patient, and clinical response rate, by patient. RESULTS: Of 1045 patients, 589 were evaluable for efficacy. At baseline, most patients had moderate or severe cough and sputum production as well as rhonchi, wheezing, and dyspnea. The microbiologic eradication rates by pathogen were 90% with once-daily cefdinir, 85% with twice-daily cefdinir, and 88% with twice-daily cefuroxime. The corresponding values for microbiologic eradication rate by patient were 90% (once-daily cefdinir), 85% (twice-daily cefdinir), and 86% (twice-daily cefuroxime). The respective clinical response rates by patient were 81%, 74%, and 80%. There were no significant differences in the incidence of drug-related adverse events or discontinuations due to adverse events. Diarrhea was the most frequent complaint. CONCLUSIONS: The results indicate that the efficacy and tolerability of cefdinir, once or twice daily, and cefuroxime were comparable with no significant differences between the regimens used. PMID- 10689212 TI - Prevalence of anti-hepatitis A antibodies in an urban middle class area of Argentina: some associated factors. AB - OBJECTIVE: This study evaluated the seroprevalence of hepatitis A virus (HAV) antibodies in 360 middle-class subjects from Buenos Aires City and its outskirts. METHODS: The study population included 360 individuals between 10 and 89 years of age, from the socioeconomic middle class in Buenos Aires City and some suburban areas of Buenos Aires province. Antibodies to hepatitis A virus were determined by enzyme immunoassay test kits. RESULTS: The overall prevalence of HAV antibodies was 42.2%. The highest percentage of seronegativity was found in the subgroup of younger people without a history of symptomatic hepatitis and living in houses with more than one bathroom (86.9%). In the subgroup aged 21 to 60 years, the highest rates of seronegativity were found in individuals with higher level of education living in houses with tap water (66.6%). In both groups, seronegativity may be correlated with a higher socioeconomic status. CONCLUSIONS: In the middle-class community studied, more than 50% of people under 30 years of age were unprotected against HAV. Thus, the use of a vaccine against hepatitis A has to be considered for the prevention of symptomatic hepatitis, especially in adults at risk of infection, such as those who travel to areas with poor sanitation, taking into consideration that the severity of the disease increases with age. PMID- 10689213 TI - Role of tumor necrosis factor-alpha in the differential diagnosis of parapneumonic effusion. AB - OBJECTIVE: This study was undertaken to investigate the role of tumor necrosis factor-a (TNF-a) in discriminating between uncomplicated parapneumonic effusion (UCPPE) and complicated parapneumonic effusion (CPPE). METHOD: Using a commercially available high sensitivity enzyme-linked immunosorbent assay (ELISA) kit, concentrations of TNF were measured in the serum (TNFs) and pleural fluid (TNFpf) of 21 patients with parapneumonic effusion (PPE), 13 patients with UCPPE, and 8 patients with nonempyemic CPPE. RESULTS: No significant difference in values of TNF concentration was found between the group with UCPPE and that with CPPE (P > 0.05). Concentration levels of TNFpf were significantly higher in the group with CPPE than in that with UCPPE (P = 0.0008). Levels of TNF in pleural fluid were significantly higher than in serum in both groups (P < 0.001). The ratio of TNF in pleural fluid to that in serum (TNFr) was significantly higher in the CPPE group than in the UCPPE group (P = 0.0002). At an optimal cutoff point of 10.7 pg/mL for TNFpf, the sensitivity was 87.5%, specificity was 92.3%, positive predictive value was 87.5%, negative predictive value was 93.3%, and total accuracy was 90.5% (P < 0.001). At an optimal cutoff point of 3.0 for TNFr, all values were 100% (P < 0.00001). CONCLUSIONS: The results of this study indicate that TNFpf, and particularly TNFr, may be helpful in discriminating between UCPPE and CPPE. However, further studies are needed in a larger population to confirm these findings. PMID- 10689214 TI - Evaluation of a supplemental assay for the diagnosis of hepatitis C virus infections. AB - OBJECTIVES: A supplemental test was evaluated for hepatitis C virus (HCV). METHODS: One hundred forty-six sera that were inconclusive or discrepant in two screening tests for HCV infection were evaluated using a supplemental test, MATRIX-HCV2 (Abbott Laboratories, Chicago, IL, USA). Results of the supplemental test were compared to the detection of HCV RNA by a nested polymerase chain reaction after a step of reverse transcription (RT-PCR). RESULTS: Thirty-nine RNA containing sera (positive with RT-PCR) of 40 (97%) reacted with at least one antigen in the supplemental test. Reactivity with one to three antigens also was observed with 77 PCR-negative sera (66%). Twenty-nine sera were found negative with both techniques. CONCLUSIONS: Despite clear results and good sensitivity, the MATRIX-HCV2 assay was poorly predictive of viremia in patients with indeterminate results in initial screening assays. PMID- 10689215 TI - Generation of reactive oxygen species and formation and membrane lipid peroxides in cells infected with Chlamydia trachomatis. AB - OBJECTIVES: Chlamydiae are obligate intracellular pathogens that cause many diseases for which the pathogenic mechanisms are largely unknown. Because reactive oxygen species (ROS) have been implicated in pathogenesis of many viral and bacterial infections, the authors assessed the release of ROS in selected host cells (monocytes, Sup-T1 cells, and Hep-2 cells) infected with Chlamydia trachomatis. METHODS: Infected cell cultures demonstrated a dramatic depletion of uric acid from culture media that was not seen in uninfected cultures. Reactive oxygen species generated in infected cultures were associated with the formation of lipid peroxides in host cell membrane. RESULTS: There was a significant increase in lipid peroxide levels in infected cells compared to uninfected controls. Ascorbic acid treatment of infected cell cultures reduced the formation of membrane lipid peroxides. CONCLUSIONS: These results suggest that ROS produced during chlamydial replication cause membrane lipid peroxidation. The role of ROS induced membrane damage in chlamydial pathogenesis is discussed. PMID- 10689216 TI - Cutaneous gnathostomiasis in a woman from Bangladesh. AB - A woman from Bangladesh who had lived in Germany for more than 2 years presented with migratory, painful swellings on her left hand and arm of 5 months duration. Laboratory examinations yielded a marked eosinophilia and a grossly elevated IgE level in combination with an inflammatory reaction restricted to the subcutaneous tissues. A preliminary diagnosis of gnathostomiasis was established and confirmed by a positive gnathostoma serology by enzyme immunoassay (EIA). Treatment was initiated with albendazole, leading to the outward migration of a larva and complete resolution of clinical disease. Currently, there is no definitive therapy that has been proved to be both safe and highly effective. A wide range of potential agents has been used in clinical studies, but only albendazole has proved to be reliably effective to date, stimulating the outward migration of larvae in a proportion of cases of cutaneous disease, as observed in the present case. PMID- 10689217 TI - Brucellosis in a mother and her young infant: probable transmission by breast milk. AB - Brucellosis, although primarily a zoonotic infection, is also a threat for human health. Infection can be transmitted to humans through direct contact with infected animals, products of conception, or animal discharges, and through consumption of potentially infected milk, milk products, or meat. Human-to-human transmission is rare. There have been case reports of transmission via blood transfusion and bone marrow transplantation from infected donors. Sexual intercourse is a possible means of transmission. Neonatal infection can be acquired transplacentally or during delivery. This report describes a mother with brucellosis who probably transmitted the infection to her 3-month-old baby by breast milk. PMID- 10689218 TI - Isolated primary hepatic lymphoma in a patient with acquired immunodeficiency syndrome. AB - Non-Hodgkin lymphoma (NHL) of the B-cell type is the second most common neoplasm in patients with human immunodeficiency virus (HIV) infection after Kaposi sarcoma (KS). The majority of cases of NHL in patients with acquired immunodeficiency syndrome (AIDS) involve extranodal sites; most frequently the gastrointestinal tract (GIT) and the central nervous system (CNS). Hepatic NHL in patients with AIDS was first described by Reichert et al in 1983 in an autopsy series. It usually presents with multiple large hepatic masses and involvement of other abdominal organs or lymph nodes. The authors present a case of primary hepatic NHL in a patient with AIDS, presenting with innumerable small intrahepatic masses without the involvement of any other organs. PMID- 10689219 TI - Editorial. AB - The cosmopolitan nature of current travel practices, as well as significant immigration from endemic areas, has led to increases in the incidence of leprosy. The classic presentation of leprosy usually appears as the indeterminate form, demonstrates hypopigmented macules with a loss of sensation. However, the manifestations can sometimes be quite protean. Sadeghi et al. review the rheumatic manifestations of leprosy with an illustrating case presenting with arthritis. Rheumatic symptoms are common in leprosy patients and may be the presenting manifestations and should be considered in patients with persistent rash and unusual arthritis who have a history of exposure to endemic areas. Significant interest has developed over the past decade in inhibiting cutaneous carcinogenesis with retinoic acid. This has been used as an effective therapy in certain genetically predisposed individuals, including individuals with a DNA repair defect in xeroderma pigmentosa. The molecular mechanisms of retinoids ability to inhibit ultraviolet light induced carcinogenesis have not been determined. Li and co-workers have examined the effect of retinoic acid on ultraviolet light induced programmed cell death (apotosis) as well as expression of the tumour suppressor gene P53. Their studies suggest that retinoic acid does not work on the initiation stages of the cancer development, but may work in the promotion and progression stage. Of more immediate clinical importance, in the Point-Counterpoint section, we have two insightful articles on how physician reimbursement affects patient care. As North American health care continues to evolve, there is constant debate on what model system works best for the ultimate benefit of our patients. Physicians, politicians, and administrators are constantly comparing the United States health care delivery to that of Canada. While no one can accurately predict the future developments in these areas, I think Dr. McElgunn sums the concerns that indeed are applicable on both sides of the border: ".socialized medical system has been of great benefit to patients but the ability of its physicians to continue to carry the system is at or near the breaking point. The ramifications of the issues of access and quality of care are harbingers of a system in turmoil." While these concerns must be dealt with, strong physician input is vital to continuing the effective evolution of our health care system. A vital part of our health care delivery is the increasing use of diagnostic tests. Key treatment decisions and interventions are based on the interpretation of these tests. However, most tests are "imperfect instruments." The article by Binder and Dreiseitl concisely reviews sensitivity, specificity, prevalence, predictive values, and likelihood ratios in a highly informative manner with significant examples. This paper provides a reference with which all physicians should be familiar. Traditionally, Western medicine has focused on a model of disease whereby pathology was regarded as well defined alteration in normal physiology that should respond to appropriate pharmaceutical or surgical interventions. However, in recent years patient focused medicine has become an important aspect of our practices. The concept of health related quality of life has represented an important advance in dealing with these concerns in our treatment of disease. Drs. Price and Harding examine the concept of health related quality of life using the example of a diabetic foot ulcer complications. These types of measures are important to understand, not only in the context of this disease but in the context of any chronic dermatologic condition. PMID- 10689220 TI - Effect of retinoic acid on apoptosis and DNA repair in human keratinocytes after UVB irradiation. AB - BACKGROUND: Skin cancer is extremely common. Epidemiological studies indicated that ultraviolet radiation (UV) is the primary cause for skin cancers, and that retinoic acid (RA) is able to inhibit this UV-induced skin carcinogenesis; however, the molecular mechanism of the anti-UV action of RA is unclear. OBJECTIVE: The purpose of this study is to investigate if RA enhances the removal of UV-induced DNA damage. METHODS: The effect of RA on UV-induced apoptosis and DNA repair was investigated by ELISA apoptosis assay and CAT assay. RESULTS: Both all-trans-RA and 9-cis-RA did not promote UV-induced apoptosis nor the repair of UV-damaged DNA in human keratinocytes. Furthermore, RA did not induce the expression of p53. CONCLUSION: The inhibition of RA on skin carcinogenesis is not due to enhanced removal of UV-damaged DNA. Therefore, RA does not inhibit skin cancer development at the initiation stage, but possibly at the promotion and progression stages. PMID- 10689221 TI - Open-label study to evaluate the healing rate and safety of the Profore Extra Four-Layer Bandage System in patients with venous leg ulceration. AB - BACKGROUND: Venous ulcers are increasing in prevalence, especially since these are observed more frequently in the elderly, and the number of individuals in this age group is becoming a larger portion of the population. OBJECTIVE: To determine the healing rate and safety of the Profore Extra Four-Layer Bandage System in the management of venous leg ulcers. METHODS: In an open-label study, patients aged 18 years or older with venous leg ulcers were treated with a high compression four-layer bandage system in which a hydrocellular dressing was placed in contact with the wound. The combination is designated the "Profore Extra Four-Layer Bandage System." Follow-up visits took place weekly unless there was heavy exudation from the ulcer or if there was marked edema of the leg at the start of the study requiring reapplication of the bandage system. RESULTS: Fifteen patients were entered into the study (men 8, women 7, mean age 66 years, mean duration of ulcers 1.3 years). Thirteen of the 15 patients completed the study, with two withdrawals. In one patient who withdrew, the ulcer became infected and required treatment with antibiotics. The other termination from the study occurred for reasons unrelated to treatment. The ulcer in this patient healed in 7 weeks. Ten of the 13 patients (77%) who completed the study, and 10 (67%) of 15, who had enrolled experienced complete (100%) healing. Healing of > 80% of the ulcers occurred in 11 of 13 patients (85%) who completed the study and in 12 (80%) of 15 enrolled patients. No patient experienced a study-related adverse event. One patient developed contact dermatitis and was later found to have stasis dermatitis. It is unclear whether the initial event was contact or stasis dermatitis. CONCLUSION: In this open-label study, a high compression system, using the Profore Extra Four-Layer Bandage with a hydrocellular dressing in contact with the wound, was found to be effective and safe for the treatment of venous leg ulcers. PMID- 10689222 TI - Perforating disorders: summary notes. AB - The intention of the Summary Notes section is to provide the practitioner and trainee with a current, concise reference source to dermatologic diseases and to serve as a form of Continuing Medical Education. Each installment will deal with a specific disease. When included, the pretest questions indicate some of the areas to be covered and will challenge your present knowledge of the material before reading further. The self-assessment post-test questions appear on page 18; the answers are on page 25. PMID- 10689223 TI - The interpretation of test results. AB - BACKGROUND: Dermatologists need to interpret an increasing number of research studies and diagnostic tests. Understanding the techniques for interpreting test results and making decisions based upon those tests represent important tools for decision making for both clinicians and researchers. OBJECTIVE: This article focuses briefly on the key parameters of diagnostic tests: sensitivity, specificity, prevalence, predictive values, likelihood ratios, and the concept of receiver-operating-characteristic (ROC) curves. A simple example is presented in a step-by-step manner. CONCLUSION: The principles of interpreting test results are easy to learn and applicable in daily clinical routine. Therefore, dermatologists should be familiar with the concepts outlined in this paper. PMID- 10689224 TI - Delay in diagnosis: indeterminate leprosy presenting with rheumatic manifestations. AB - BACKGROUND: Rheumatic complications are common in leprosy (Hansen's disease) and can be the primary complaint delaying accurate diagnosis. OBJECTIVE: Such a case is reported here: a 61-year-old woman with indeterminate leprosy presented with symmetric arthritis and purpura. Despite biopsy and evaluation by several physicians, leprosy was not suspected. After 2 years of progressive symptoms, a second biopsy revealed lepromatous leprosy. CONCLUSION: In this case, lack of clinical suspicion and unfamiliarity with the histology of indeterminate leprosy delayed diagnosis and treatment. Leprosy should be considered in the differential diagnosis of patients presenting with unusual rheumatic and persistent cutaneous manifestations. PMID- 10689225 TI - Lichen planopilaris-like changes arising within an epidermal nevus: does this case suggest clues to the etiology of lichen planopilaris? AB - BACKGROUND: Lichen planopilaris shows a perifollicular lymphocytic infiltrate at the level of infundibulum and the isthmus of the hair bulge resulting in necrotic changes within keratinocytes and eventually hair loss. OBJECTIVE: We present a 14 year-old black male with a history of a raised epidermal lesion on the scalp that was present at birth. Over the past few years, the patient developed gradual hair loss and increased verrucous changes of the skin within the original lesion. Histologic sections of the area showed features consistent with an epidermal nevus peripherally, with a central area showing some features characteristic of those seen in lichen planopilaris. In addition, there was marked hyperkeratosis with increased yeast and bacteria within the follicles. CONCLUSIONS: Initiating factors in lichen planopilaris are not well defined. Overgrowth of microorganisms with hyperkeratosis results in factors that disrupt the immune privilege of the hair follicle, leading to an immunologic reaction that is limited to the follicle and spares surrounding eccrine structures. This case suggests possible mechanisms involved in the induction of lichen planopilaris. PMID- 10689226 TI - Treatment of vascular lesions in pigmented skin with the pulsed dye laser. AB - BACKGROUND: Vascular lesions occurring in African-American patients are often not treated because of the risk of local side effects. OBJECTIVE: The study was to determine the efficacy of the flashlamp-pumped dye (FLPD) laser in the treatment of vascular malformations in African-American patients. METHODS: All lesions in three patients were treated with the FLPD laser using a 585 nm wavelength, 5 mm spot size, 450 pulse width, and fluences ranging from 6.5 to 8.5j per cm2. CONCLUSION: The FLPD was effective in treating vascular malformations. Transient changes in colour and skin texture occurred at the treated sites. PMID- 10689227 TI - Wrinkling due to mid-dermal elastolysis: two cases and literature review. AB - BACKGROUND: Mid-dermal elastolysis is an acquired disorder of elastic tissue clinically characterized by diffuse fine wrinkling, most often of the trunk and arms. Histologically, a clear band of elastolysis is present in the mid-dermis. OBJECTIVE: Although examples of diffuse elastolysis are well known, only a small number of patients with mid-dermal elastolysis have been reported to date. We present two patients with clinical and histological evidence of mid-dermal elastolysis, review the literature, and summarize the salient features of some common disorders of elastic tissue. METHODS: The first patient presented with fine wrinkles and papules over the upper arms, upper chest, and axillae, and demonstrated increased laxity of the eyelids. The second patient had striking wrinkles extending in a band-like pattern on her arms, upper chest, back, and abdomen. Neither one of our patients had a previous history of skin inflammation, urticaria, or any other underlying diseases related to their skin changes. Skin biopsies were taken from lesional and perilesional skin of both patients, and were stained with hematoxylin and eosin, and with elastic tissue stain. In addition, a tissue sample from Patient 1 was fixed for electron-microscopy. RESULTS: Hematoxylin and eosin stains did not demonstrate specific changes or diagnostic patterns. However, elastic tissue stains revealed a band-like loss of elastic tissue in the mid-dermis. Elastic tissue in the remaining superficial and deep dermis stained normally. Electron-microscopy was consistent with these findings and revealed significant loss of elastic tissue limited to the mid dermis. CONCLUSION: We have presented two cases of mid-dermal elastolysis and reviewed the literature. To date, the pathophysiology of mid-dermal elastolysis had not been elucidated and no definitive therapy exists. PMID- 10689228 TI - The impact of foot complications on health-related quality of life in patients with diabetes. AB - BACKGROUND: The concept of health-related quality of life (HRQoL) has been the focus of much debate in recent years. However, within diabetes the focus has centred on the behavioural adaptation to a chronic disease state. The impact of foot complications is witnessed regularly in the clinical setting; amputation in this group is usually preceded by ulceration and a worsening cycle of foot problems. OBJECTIVE: This review sets out to investigate the literature on foot complications in those with diabetes, to assess the cost to the individual in terms of impact on everyday living. CONCLUSION: The literature on the specific impact of foot complications is limited, but indicates a situation in which those with diabetic foot ulceration may have an even poorer HRQoL than those who have experienced an amputation related to diabetes. In order to assess the full impact of new treatments or therapeutic interventions, it is vital that further research is conducted in this area. PMID- 10689229 TI - Sporotrichosis infection on mines of the Witwatersrand. AB - BACKGROUND: Our knowledge of skin disease is often based on fortuitous situations that have provided opportunities for observation and study of the disease. One of these diseases is sporotrichosis. OBJECTIVE: This article examines a symposium published in 1947 by the Transvaal Chamber of Mines in South Africa. It reviews the approach taken in the investigation of a large outbreak of sporotrichosis in the mine workers. CONCLUSION: The investigation of this outbreak has contributed significantly to our present day knowledge of sporotrichosis, the causative organism, its mode of spread, and its clinical features. It is also a striking example of meticulous scientific research and observation. PMID- 10689230 TI - International regulations for automobile driving and epilepsy. AB - BACKGROUND: Many patients with epilepsy travel abroad and drive automobiles with the assumption that policies, rules, and regulations on epilepsy and driving are similar to those of their home countries. This paper investigates the driving restrictions and other pertinent information on this issue in foreign countries. METHODS: A questionnaire was sent to 231 neurologists (chosen from American neurological and epilepsy societies) from 84 countries and to 230 official (embassies and consulates) representatives of 134 countries asking for the local rules and regulations and their comments on driving and epilepsy. RESULTS: One hundred and sixty-six responses were received from 96 of 134 (72%) countries. One hundred and six neurologists (of 231 queried [46%]) responded. In 16 countries, persons with epilepsy are not permitted to drive. In the remaining countries, these patients must have a seizure-free period of 6 to 36 months. This period varies according to the type of seizure. In five countries, physicians must report the names of these patients to their local authorities. In many countries, the rules and regulations are being reevaluated and changed. CONCLUSIONS: Patients with epilepsy who plan to drive overseas are advised to contact local embassies and consulates, well before their trips (and keep records of the communications) to obtain the latest information on the rules and regulations governing the driving of automobiles in those countries. PMID- 10689231 TI - Neuropsychiatric problems in 2,500 long-term young travelers to the tropics. AB - BACKGROUND: The prevalence and features of travel associated neuropsychiatric problems (NPP) and their relation to previous psychological consultations, antimalarials and recreational drug use have not been adequately studied. METHODS: A two-phase postal and telephone survey has been conducted among 2,500 young travelers to tropical countries. We measured the rate and duration of NPP, characterized their features, and their association with previous psychological profiles, itinerary, type of travel, consumption of recreational drugs, and malaria prophylaxis. RESULTS: First phase: Out of 1,340 respondents, 151 (11.3%) indicated that they had NPP during travel, in contrast with 2.3% who needed psychological consultation before travel (p<.001). Second phase: 117 of 151 responded to the study questionnaire. The mean age of the respondents was 24.4 years, 54.7% were female, and the mean stay abroad was 5.3 months. The most common NPP were sleeping disturbances (52.1%), fatigue (48.7%) and dizziness (39.3%). Thirty-three travelers (2.5%) had severe symptoms, and 16 (1.2%) had symptoms lasting more than 2 months. Seven travelers had pure or mixed depressive symptoms. Consumption of recreational drugs was admitted by 22.2%. Mefloquine was used significantly more often by those who suffered NPP, than by the entire cohort (98.2% vs. 70.7%; p<.001). CONCLUSIONS: Long-term travel to the tropics was associated, in this cohort, with a considerable rate of neuropsychiatric symptoms. The majority of the responding travelers were females, used mefloquine as prophylaxis, and at least one fifth used recreational drugs. PMID- 10689232 TI - Survey of rabies preexposure and postexposure prophylaxis among missionary personnel stationed outside the United States. AB - BACKGROUND: Of the 36 cases of human rabies that have occurred in the United States since 1980, 12 (33%) were presumed to have been acquired abroad. In the United States, it is recommended that international travelers likely to come in contact with animals in canine rabies-enzootic areas that lack immediate access to appropriate medical care, including vaccine and rabies immune globulin, should be considered for preexposure prophylaxis. In 1992, the death of an American missionary who had contracted rabies while stationed in Bangladesh highlighted this high-risk group. METHODS: To assess their knowledge of rabies risk, rabies exposures, and compliance with preventive recommendations, we asked 695 missionaries and their family members to complete questionnaires about their time stationed abroad. RESULTS: Of the 293 respondents stationed in countries where rabies is endemic, 37% reported prior knowledge of the presence of rabies in their country of service. Only 28% of the personnel stationed in rabies-endemic countries received preexposure prophylaxis. Having preexposure prophylaxis specifically recommended increased the likelihood of actually receiving it (O.R. 15.6, 95%CI 7.4 - 34.9). There were 38 reported exposures (dogs = 66%, another human = 20%), proven or presumed to be rabid. Three of the people exposed received rabies immune globulin and vaccine; 11 received vaccine alone; 8 received only basic first aid, and 16 received no treatment. CONCLUSIONS: Although American missionaries stationed abroad are at an increased risk for exposure to rabies, compliance with established preventive measures was low. Prior to being stationed abroad, an educational rabies-prevention briefing, including encouragement to receive preexposure prophylaxis, could be an effective intervention for missionaries to decrease their risk of rabies. PMID- 10689233 TI - Etiology of travelers' diarrhea on a Caribbean island. AB - BACKGROUND: Between December 6, 1994 and March 10, 1996, a study of the etiology of diarrhea was carried out among 332 travelers to five all-inclusive hotels in Negril, Jamaica. METHODS: Stool specimens were collected and sent to Montego Bay for laboratory analysis. Escherichia coli strains isolated at the Jamaican laboratory were sent to Houston for toxin testing. RESULTS: A recognized enteropathogen was found in 118 of the 332 (35.5%) cases. Enterotoxigenic E. coli (ETEC) were the most commonly identified pathogen (87/332; 26.2%) followed by Salmonella (4.2%) and Shigella (4.2%). Clustering of etiologically defined cases was studied at each hotel. A cluster was defined as 2 or more cases with the same pathogen identified in the same hotel within 7 days. In the 3 hotels with the highest number of cases of diarrhea, enteropathogens were part of a cluster in 65 of 99 cases (65.7%) of diarrhea of which an etiologic agent was identified. In the other 2 hotels, only 4 of 20 cases (20%) occurred in clusters. CONCLUSIONS: A total of 25 clusters of travelers' diarrhea cases was detected at the five hotels during the study period. Seventeen of 25 (68%) ETEC isolations occurred as part of a clustering of diarrhea cases. The largest outbreak of pathogen-identified diarrhea consisted of 7 cases of ETEC producing both heat-stable and heat-labile enterotoxins. In the Jamaican hotels with all inclusive meal packages most diarrhea cases occurred as small clusters, presumably as the result of foodborne outbreaks. PMID- 10689234 TI - Typhoid fever in group travelers: opportunity for studying vaccine efficacy. AB - BACKGROUND: Typhoid fever (TF) is a rare disease among travelers to endemic areas, and little is known about its travel-related epidemiology. In addition, efficacy data on TF vaccines in travelers is scanty. During 3 months of 1994/95, six cases of TF were reported in The Netherlands among participants of four package tours to Indonesia provided by the same operator. The present study was designed to describe the epidemiology of TF in these groups, and to assess whether travel groups can be used for studying the efficacy of TF vaccines in travelers. METHOD: Questionnaire-based historical cohort study of participants of 4 groups that stayed in the same hotels along their tours (n=156). TF was defined as blood culture-confirmed Salmonella typhi infection. Submitted isolates were typed by antigen and phage typing. Immunization status was considered documented if ascertained by written records. RESULTS: Among 110 participants (71%), six cases of TF were identified (group specific attack rate AR 5.4%), three of which were from one travel group (AR 12.0%). There were no significant differences by age or sex. Three submitted S. typhi isolates showed three different types, two of which were in the same group. Eighty-three percent of respondents reported documented TF vaccination in the preceding 3 years. All cases occurred in recipients of the oral Ty21a vaccine (AR 10.2%, 95% CI 3.8-20.8%), but differences with nonvaccinees and recipients of the heat-inactivated whole cell or Vi-antigen polysaccharide vaccines were not significant. CONCLUSIONS: Although TF is rare in travelers, infections with different strains of S. typhi can occur in one travel group. Travel groups offer an opportunity for retrospective assessment of vaccine efficacy, provided that equal chance of exposure is largely guaranteed; case ascertainment is maximally specific and similar in the vaccine groups; vaccine status is ascertained accurately; and prior immunity by previous exposures to and use of antibiotics effective against the infection are excluded from, or controlled for in, the analysis. PMID- 10689235 TI - Experience of corporate medical assistance clinics during the Centennial Olympic Games, Atlanta, 1996. AB - The health of travelers returning home from developing countries has received increased attention in recent years. Much of this attention has centered on immunizations, malaria chemoprophylaxis and treatment of traveler's diarrhea. In contrast, there are very few data on the health problems of international travelers to developed countries such as the United States. We studied the experience of two corporate medical assistance clinics established for both national and international travelers to Atlanta, Georgia during the Centennial Summer Olympic Games in 1996. PMID- 10689237 TI - Female genital schistosomiasis. AB - Schistosoma haemtobium infection in travelers from endemic areas is usually asymptomatic, or presents with hematuria. Uncommon manifestations include neurological syndromes, genital dysaesthesias and watery or blood stained semen. This organism also causes disease within all structures of the female genital tract because of communications between pelvic venous complexes, and can occur long after return home. Schistosomiasis may not be suspected, resulting in delays in diagnosis and treatment. We present two cases which illustrate the diverse nature of this condition. PMID- 10689236 TI - Double-blind, randomized, placebo controlled pilot study evaluating efficacy and reactogenicity of an oral ETEC B-subunit-inactivated whole cell vaccine against travelers' diarrhea (preliminary report). AB - Diarrhea caused by enterotoxigenic E.coli (ETEC) is an important health problem in developing countries and in travelers to these areas. In previous trials formulations of ETEC vaccines containing the B-subunit of cholera toxin, which is antigenically similar to the heat labile enterotoxin of ETEC, and the most prevalent colonization factor antigens of ETEC, were shown to stimulate relevant mucosal immune responses in volunteers from Sweden and Egypt. PMID- 10689238 TI - Tropical sprue after travel to Tanzania. AB - Tropical sprue (TS) is a diagnosis to consider in travelers with prolonged diarrhea and a malabsorption syndrome after return from tropical countries, particularly India and Southeast Asia. TS is an unusual condition in tropical Africa. Textbooks of tropical medicine indicate a low endemicity in Nigeria and a limited number of cases in South Africa and Zimbabwe. A Medline search from 1979 to mid 1998 using "Tanzania and tropical sprue" as key words disclosed no hits. We report herein a case of TS in a European traveler, who lived in Tanzania for 8 months. PMID- 10689239 TI - Reactive arthritis associated with typhoid vaccination in travelers: report of two cases with negative HLA-B27. AB - As international travel to developing countries increases, more people seek medical advice concerning food and water-borne diseases, including typhoid fever. Prevention of typhoid fever in high-risk groups (travelers to endemic areas, laboratory workers and household contacts of typhoid carriers) should rely primarily on prevention of exposure. However, immunization is an important adjunct. The decision to immunize against typhoid fever should be individualized, taking into account the benefits versus the risk of possible adverse reactions. Cases of reactive arthritis have been associated with the heat-phenol inactivated 'whole cell' parenteral vaccine, but to our knowledge reactive arthritis has not been previously reported with the oral form (Ty21a). This is a report of HLA-B27 negative reactive arthritis occurring in two travelers after the administration of oral Ty21a typhoid vaccine. PMID- 10689240 TI - The Japanese need travel vaccinations. AB - At a travel clinic in Kathmandu we reviewed the vaccination records from March 1997 to March 1998 for all travelers to developing countries like Nepal, for two important vaccines, namely, typhoid and hepatitis A. These travelers visited the clinic for various medical problems. One of the reasons for doing this study was that in previous years we saw a disproportionate number of Japanese travelers with hepatitis A, who had not taken the hepatitis A vaccine or immune gamma globulin for prevention of this illness. We hypothesized, therefore, that one of the reasons that Japanese patients visiting our clinic had higher rates of hepatitis A was because they were not vaccinated against this disease. There were 765 tourists for that time period out of which about 10% were Japanese. The rest were Americans, British, Israelis, Canadians, Australians, Danish and a small miscellaneous group from other countries. PMID- 10689241 TI - Safety of iodine based water sterilization for travelers. AB - The recent report by Khan et al. of an unexpectedly high concentration of free iodine in water filters, which may have led to the high proportion of abnormal thyroid function tests in Peace Corps workers, is of concern for travel advisors when asked to recommend suitable means of water sterilization. Many travelers use iodine based filters and/or chemicals for purification of water when traveling in areas with contaminated water supplies and may therefore be at risk of excess iodine intake. Aside from iodine impregnated resin filtration systems, tetraglycine hydroperiodide tablets, tincture of iodine 2% and more commonly, chlorine-based proprietary products are widely used to sterilize water for drinking, and usually purchased by travelers without advice on how they should be used. A single tetraglycine hydroperiodide tablet in a liter of water releases 8 mg of iodine in comparison to the 10 mg/liter released from the iodinated resin pumps described by Khan et al. Although the instructions for using iodine tincture are imprecise, the normal recommendation is 5 drops per liter of water, increasing this to 12 drops where Giardia cysts may be present. The lower of the two doses would yield about 2 mg/liter of free iodine per liter depending on the pipette used, although, because of the potassium iodide present in the formulation, a total of 4 mg iodine would be available for absorption. PMID- 10689242 TI - Intractable nausea, vomiting and diarrhea in a Mexican woman with No recent travel history. AB - A 45-year-old Mexican woman with a history of noninsulin dependent diabetes mellitus (NIDDM), hypertension, and coronary artery disease presented to the hospital after 2 months of intractable nausea, vomiting and diarrhea-all made worse by eating and drinking. She reported fever, chills, anorexia and a documented 50-pound weight loss during this period. She denied the signs and symptoms of melena, hematochezia, steatorrhea or constipation. She also reported left leg pain and decreased sensation and strength of her left leg compared to the right leg. She had been hospitalized 2 weeks prior to admission with the same symptoms and a diagnosis of viral gastroenteritis. She was also treated for H. pylori, but subsequent biopsy results were negative by Steiner stain. PMID- 10689243 TI - Two simultaneous cases of Cyclospora cayatensis enteritis returning from the Dominican Republic. AB - According to the "International Passenger Survey," published in 1996 by the Office of Trading Standards, 534,000 British people traveled to the Caribbean area (personal communication, ABTA, 1998). The Dominican Republic, the eastern end of a large Caribbean island, has become in recent years one of the most popular destinations for UK holidaymakers as well as for travelers from many other countries. Cyclospora cayatensis has been firmly identified as a cause of gastroenteritis among international travelers,1 including human immunodeficiency virus (HIV)-positive individuals,2 but it has not been previously reported in the literature in British individuals returning from this increasingly popular vacation destination. PMID- 10689244 TI - Sea urchin puncture resulting in PIP joint synovial arthritis: case report and MRI study. AB - Of the 600 species of sea urchins, approximately 80 may be venomous to humans. The long spined or black sea urchin, Diadema setosum may cause damage by the breaking off of its brittle spines after they penetrate the skin. Synovitis followed by arthritis may be an unusual but apparently not a rare sequel to such injury, when implantation occurs near a joint. In this case report, osseous changes were not seen by plain x-rays. Magnetic resonance imaging (MRI) was used to expose the more salient features of both soft tissue and bone changes of black sea urchin puncture injury 30 months after penetration. In all likelihood, this type of injury may be more common than the existing literature at present suggests. It is believed to be the first reported case in this part of the world as well as the first MRI study describing this type of joint pathology. Local and systemic reactions to puncture injuries from sea urchin spines have been described previously. These may range from mild, local irritation lasting a few days to granuloma formation, infection and on occasions systemic illness. The sea urchin spines are composed of calcium carbonate with proteinaceous covering. The covering tends to cause immune reactions of variable presentation. There are only a handful of reported cases with sea urchin stings on record, none of them from the Red Sea. However, this condition is probably more common than is thought and can present difficulty in diagnosis. In this case report, the inflammation responded well to heat treatment, mobilization and manipulation of the joint in its post acute and chronic stages. As some subtle changes in soft tissues and the changes in bone were not seen either on plain x-rays or ultrasound scan, gadolinium-enhanced MRI was used to unveil the marked changes in the joint. PMID- 10689245 TI - A case of dengue from Pune, India. AB - Dengue is an acute infectious disease, caused by a single stranded ribonucleic acid (RNA) virus of the family of Flaviviridae, genus Flavivirus, transmitted by Aedes mosquitos, the most important vectors being Aedes aegypti and Aedes albopictus. There are four serotypes, DEN-1, DEN-2, DEN-3, DEN-4 that are nearly human specific. The geographical distribution of dengue is pantropical, except for Madagascar and some African regions. It is endemo-epidemic in tropical and subtropical countries included between 25 degrees north latitude and 25 degrees south latitude, particularly in southeast Asia and epidemic in the Caribbean, West-Africa, tropical American and Pacific Islands. Further spread will depend on the invasion of Aedes aegypti into new areas in South America. Maintenance of dengue is supported by an increasing demography, uncontrolled urbanization and climatic conditions favorable to the vectors. Spread of dengue is primarily caused by modern transportation, especially aircraft. No vaccine and no specific treatment exist so that mosquito control is the only way to reduce the incidence of dengue around the world. Dengue is, obviously, an important risk for travelers going to endemic areas. PMID- 10689246 TI - Indigenous malaria in a suburb of Ghent, Belgium. AB - We report here details of a patient with Plasmodium falciparum malaria which was acquired in the vicinity of Ghent (Evergem) in July 1997. Indigenous malaria disappeared from Belgium in 1938. Due to an increase in international travel, the influx of migrant labor and the changing environmental conditions, there has been an upsurge of imported malaria. Airport- and port-malaria is acquired through the bite of a tropical anophelline mosquito by people whose geographical history excludes exposure to this vector in its natural habitat. As far as we know, only two cases of port-malaria have been reported: in Marseille. We describe here another possible case of port-malaria due to infection with P. falciparum in a 42 year-old woman with an underlying non-Hodgkin lymphoma. PMID- 10689247 TI - Prevalence of nonfatal coronary heart disease among American adults. AB - BACKGROUND: Few national estimates of the prevalence of coronary heart disease in the United States are available. METHODS: By using data from the Third National Health and Nutrition Examination Survey (1988 to 1994), we estimated prevalence of angina pectoris by questionnaire, self-reported myocardial infarction, and electrocardiographically (ECG)-defined myocardial infarction. RESULTS: Among participants aged >/=40 years who attended the medical examination, the age adjusted prevalence of angina pectoris, self-reported myocardial infarction, and ECG-defined myocardial infarction were 5.8% of 9255, 6.7% of 9250, and 3.0% of 8206 participants, respectively. Among participants aged >/=65 years compared with those aged 40 to 64 years, the prevalence of a self-reported myocardial infarction was more than 3 times higher and that of ECG-defined myocardial infarction more than 4 times higher. The prevalences of self-reported myocardial infarction and ECG-defined myocardial infarction, but not angina pectoris, were higher among men than women. Among women, prevalence of angina pectoris and self reported myocardial infarction were highest among blacks; among men, these coronary heart diseases were somewhat higher among whites. Prevalence of ECG defined myocardial infarction were similar for all 3 race or ethnicity groups in either sex. The age-adjusted prevalence of coronary heart disease defined by the presence of any of these conditions was 13.9% among men and 10.1% among women. CONCLUSIONS: Although the management of coronary heart disease has improved during the past 2 decades, it remains an important prevalent disease burden among adults. PMID- 10689248 TI - Three-dimensional echocardiographic assessment of annular shape changes in the normal and regurgitant mitral valve. AB - OBJECTIVES: To compare mitral annular shape and motion throughout the cardiac cycle in patients with normal hearts versus those with functional mitral regurgitation (FMR). BACKGROUND: The causes of mitral regurgitation without valvular disease are unclear, but the condition is associated with changes in annular shape and dynamics. Three-dimensional (3D) imaging provides a more comprehensive view of annular structure and allows accurate reconstructions at high spatial and temporal resolution. METHODS: Nine normal subjects and 8 patients with FMR undergoing surgery underwent rotationally scanned transesophageal echocardiography. At every video frame of 1 sinus beat, the mitral annulus was manually traced and reconstructed in 3D by Fourier series. Annular projected area, nonplanarity, eccentricity, perimeter length, and interpeak and intervalley spans were determined at 10 time points in systole and 10 points in diastole. RESULTS: The mitral annulus in patients with FMR had a larger area, perimeter, and interpeak span than in normal subjects (P <.001 for all). At mid-systole in normal annuli, area and perimeter reach a minimum, nonplanarity is greatest, and projected shape is least circular. These cyclic variations were not significant in patients with FMR. Annular area change closely paralleled perimeter change in all patients (mean r = 0.96 +/- 0.07). CONCLUSIONS: FMR is associated with annular dilation and reduced cyclic variation in annular shape and area. Normal mitral valve function may depend on normal annular 3D shape and dimensions as well as annular plasticity. These observations may have implications for design and selection of mitral annular prostheses. PMID- 10689249 TI - When not doing tests is the right thing to do. PMID- 10689250 TI - Increase in ST-segment elevation immediately after reperfusion: cause and meaning. PMID- 10689251 TI - Cardiology, for what it's worth. PMID- 10689252 TI - Efficient utilization of echocardiography for the assessment of left ventricular systolic function. AB - BACKGROUND: We hypothesized that patients could be selected for echocardiographic evaluation of left ventricular (LV) systolic function on the basis of historic, clinical, radiographic, and electrocardiographic criteria. METHODS AND RESULTS: We prospectively evaluated 300 consecutive inpatients referred for the echocardiographic assessment of LV function, of whom 124 (41%) had LV systolic dysfunction (LVSD) (LV ejection fraction <0.45). Among the historic variables, male sex was the only predictor of LVSD, whereas of the abnormal physical and radiographic findings, cardiomegaly on chest radiography was the only predictor. Among the electrocardiographic findings, the presence of left bundle branch block was positively correlated with the presence of LVSD, whereas a normal electrocardiogram was negatively correlated with this finding. Only 2 patients with LVSD had a normal electrocardiogram. The addition of significant predictors on physical examination and chest radiography doubled the predictive value of the historic variables for determining LVSD. The addition of electrocardiographic findings further doubled the predictive value of the model. Almost 45% of the predictive power of the final multivariate model (chi-square of 48 of the total chi-square of 108) was based on the absence of normal electrocardiogram in patients with LVSD. When chest radiographic findings were excluded from the model, the overall predictive power of the model did not change, with the normal electrocardiogram gaining greater prominence: Full 56% of the predictive power of the model (chi-square of 60 of the total chi-square of 108) resided in the ability of a normal electrocardiogram to discriminate between patients with and those without LVSD. CONCLUSIONS: Historic, chest radiographic, and electrocardiographic variables can be used to predict low likelihood of LVSD on echocardiography. In particular, when the electrocardiogram is normal, it is extremely unlikely to have LVSD. It can be argued that such patients should not be referred for echocardiography. PMID- 10689253 TI - Infusion versus bolus contrast echocardiography: a multicenter, open-label, crossover trial. AB - BACKGROUND: In current practice, contrast echocardiography is performed with single or multiple bolus injections, which often result in an uncontrolled period of attenuation followed by transient left ventricular opacification (LVO). Because a "slow bolus" appears to reduce attenuation and prolong LVO, we hypothesized that a controlled infusion of contrast might provide a more uniform contrast effect with less attenuation and longer contrast duration. METHODS AND RESULTS: We sought to test the hypothesis by using an infusion of contrast (DEFINITY [perflutren], The DuPont Pharmaceuticals Co, Medical Imaging, North Billerica, Mass) that is stable when diluted in saline in a randomized, multicenter, controlled, crossover trial. Sixty-four patients with poor noncontrast images were recruited at 3 centers and randomly assigned to 2 single "slow" bolus injections of contrast (10 microL/kg each over a period of 30 to 60 seconds) or an infusion (1. 3 mL in 50 mL normal saline initially at 4.0 mL/min) of contrast. Patients then returned within 24 to 72 hours for the alternative form of contrast delivery. Three independent experienced echocardiographers viewed 30 seconds of videotape for all optimal baseline and optimal contrast images to score LVO and qualitatively assessed endocardial border evaluability. The duration of adequate LVO then was independently assessed by review of the entire videotape. Three independent sonographers traced single-frame, digitally captured images to measure the length of the contiguous endocardial border visualized. Both bolus and infusion administration demonstrated improved LVO (>90% by all blinded readers, P <.01) and endocardial border visualized (mean increase of 1.8 to 4.7 cm at both end-diastole and end-systole, all P <.05) as compared with baseline images. However, contrast infusion resulted in a longer duration of LVO (range of mean durations for each reader, 158 to 174 seconds longer, P <.05) and a shorter duration of attenuation (18 to 54 seconds, P <.05) compared with either bolus injection. There were no severe adverse events with contrast infusion. CONCLUSIONS: Contrast echocardiography delivered as an infusion optimizes the contrast effect by decreasing the attenuation period, extending the LVO duration, and providing a uniform contrast effect that may be useful in obtaining multiple echocardiographic views, stress echocardiography, myocardial perfusion imaging, and applications in which blood flow must be quantified. PMID- 10689254 TI - Effects of mental stress on brachial artery flow-mediated vasodilation in healthy normal individuals. AB - BACKGROUND: Mental stress is associated with increased risk for cardiovascular events, possibly because of acute increases in endogenous catecholamines. Recently, brachial artery flow-mediated vasodilation has been used for noninvasive assessment of macrovascular endothelial function. The effect of mental stress and its associated changes in sympathetic activation on brachial artery endothelium-dependent vasomotor tone in vivo remains unknown. METHODS AND RESULTS: Two-dimensional ultrasound was used to measure brachial artery flow mediated vasodilation before and after mental stress (provoked by a standard arithmetic challenge) in 21 healthy individuals (10 men, 11 women; average age 23.5 years). The flow stimulus resulted from a 3-minute cuff occlusion of distal forearm blood flow, causing distal hyperemia and a transient 2- to 3-fold increase in brachial artery blood flow on cuff release. During mental stress, heart rate increased on average by 29.6% and blood pressure increased on average by 17.9%. The sympathetic stimulus resulted in a 64% average increase in flow mediated vasodilator response (P <.001). The enhanced vasodilator response during mental stress was similar for men and women. CONCLUSIONS: Mental stress can have marked effects on endothelium-dependent, flow-mediated vasodilation in healthy, normal individuals. Similar studies in individuals with impaired endothelial function may further our understanding of the role of mental stress in the development of cardiovascular events. PMID- 10689255 TI - Interpretation of echocardiographic measurements: a call for standardization. AB - BACKGROUND: Although echocardiography is used extensively in clinical medicine, guidelines for quantitative interpretation of echocardiographic measurements are unavailable. The goals of this investigation were to provide an overview of scientific standards for formulating reference values, with clinical chemistry used as a model, to evaluate published echocardiographic reference limits, to survey clinical echocardiography laboratories regarding their interpretation of echocardiographic measurements, and to provide recommendations for improving the interpretation and reporting of echocardiographic measurements. METHODS AND RESULTS: We reviewed the original reports of the International Federation of Clinical Chemistry on guidelines for formulating reference values. We obtained published reports on echocardiographic reference limits through searches of electronic databases supplemented by a manual search of relevant bibliographies. We also surveyed echocardiographic laboratories in 35 adult acute-care hospitals in Eastern Massachusetts. Studies on echocardiographic reference values were evaluated with the use of guidelines from clinical chemistry. Responses from the 29 participating echocardiographic laboratories were evaluated for their practice of quantitative echocardiographic interpretation. There is considerable heterogeneity in the echocardiographic reference values available in the literature. There is also a lack of agreement in the literature and among echocardiographers regarding the partitioning of reference values (by sex, ethnicity, or age), the anthropometric measure to be used for indexation, and the choice of cut-points for categorizing values within the abnormal range. CONCLUSIONS: We advocate that echocardiographic reference limits be standardized and a consensus generated regarding the partitioning of reference limits and the indexation of echocardiographic measurements. Such measures can aid in quantitative echocardiographic interpretation and render the results more scientific and consistent. PMID- 10689256 TI - Will the use of low-molecular-weight heparin (enoxaparin) in patients with acute coronary syndrome save costs in Canada? AB - BACKGROUND: One-year follow-up data from the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-Wave Coronary Events (ESSENCE) trial show that use of low molecular-weight heparin (enoxaparin) compared with unfractionated heparin in patients hospitalized with unstable angina or non-Q-wave myocardial infarction is associated with a 10% reduction in the cumulative 1-year risk of death, myocardial infarction, or recurrent angina. Given the higher acquisition cost of enoxaparin relative to unfractionated heparin, we assessed whether the reduced use of revascularization procedures and related care makes enoxaparin a cost saving therapy in Canada. METHODS AND RESULTS: We analyzed cumulative 1-year resource use data on the 1259 ESSENCE patients enrolled in Canadian centers (40% of the total ESSENCE sample). Patient-specific data on use of drugs, diagnostic cardiac catheterization, percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, and hospital days were available from the initial hospital stay and cumulative to 1 year. Hospital resources were costed with the use of data from a teaching hospital in southern Ontario that is a participant in the Ontario Case Costing Project. During the initial hospital stay, use of enoxaparin was associated with reduced use of diagnostic catheterization and revascularization procedures, with the largest effect being reduced use of percutaneous transluminal coronary angioplasty (15.0% vs 10.6%; P =.03). At 1 year, the reduced risk and costs of revascularization more than offset increased drug costs for enoxaparin, producing a cost-saving per patient of $1485 (95% confidence interval $-93 to $3167; P =.06). Sensitivity analysis with lower hospital per diem costs from a community hospital in Ontario still predicts cost savings of $1075 per patient over a period of 1 year. CONCLUSIONS: The acquisition and administration cost of enoxaparin is higher than for unfractionated heparin ($101 vs $39), but in patients with acute coronary syndrome, the reduced need for hospitalization and revascularization over a period of 1 year more than offsets this initial difference in cost. Evidence from this Canadian substudy of ESSENCE supports the view that enoxaparin is less costly and more effective than unfractionated heparin in this indication. PMID- 10689257 TI - A comparison of electrocardiographic changes during reperfusion of acute myocardial infarction by thrombolysis or percutaneous transluminal coronary angioplasty. AB - BACKGROUND: Different electrocardiographic changes have been described during thrombolytic therapy for acute myocardial infarction to indicate successful reperfusion. The occluded coronary artery also can be reopened by percutaneous transluminal coronary angioplasty (PTCA). This study was performed to compare electrocardiographic changes during primary or rescue PTCA and thrombolytic therapy. The electrocardiographic changes were studied directly at the moment of reperfusion during PTCA. METHODS AND RESULTS: Continuous 12-lead electrocardiographic monitoring was performed in 110 patients with acute myocardial infarction undergoing a reperfusion intervention (thrombolytic therapy or primary or rescue PTCA) to assess electrocardiographic changes during reperfusion. Patency and Thrombolysis In Myocardial Infarction flow in the infarct-related artery were assessed by coronary angiography. During reperfusion of the infarct-related coronary artery, early signs of reperfusion were an increase of ST-segment deviation (30%), ST-segment normalization (70%), and terminal T-wave inversion (60%); only 11% of patients showed no ST-segment changes. Thrombolytic therapy was significantly more often accompanied by a transient increase in ST-segment deviation compared with primary PTCA. Accelerated idioventricular rhythm was documented in 51%, an increase in the number of ventricular premature complexes in 42%, nonsustained ventricular tachycardia in 7%, and bradycardia in 18% of all patients. CONCLUSIONS: This study confirms the occurrence of specific electrocardiographic changes at the time of reperfusion. The pattern of ST-segment change upon reperfusion relates to the type of treatment. Awareness of electrocardiographic changes at the moment of reperfusion will help to select patients for rescue PTCA and can be used to assess the effect of future pharmacologic interventions to limit reperfusion damage. PMID- 10689258 TI - Stents covered by autologous venous grafts: feasibility and immediate and long term results. AB - BACKGROUND: Previous experimental studies with a new covered stent, the autologous venous graft-covered stent (AVGCS), have shown favorable results. The aim of this study was to evaluate the feasibility and safety of this new technique in human coronary arteries and to compare the long-term outcome with uncovered stents. METHODS AND RESULTS: A venous graft was removed from an upper limb. A conventional stent then was covered by the venous graft. Fifty-eight AVGCS were implanted in 56 patients, including 16 patients with acute coronary syndromes (ACS). Additionally, in 114 patients, 138 uncovered stents were implanted, serving as a control group, including 38 patients with ACS. The procedure was successful in all patients. Stent thrombosis was observed in 3 patients in the control group and in 1 patient with an AVGCS. There was a trend for the minimal luminal diameter to be greater in the AVGCS group at follow-up (P =.07), and statistical significance was observed in patients with ACS (P <.01). The target vessel revascularization and the restenosis rates were similar between the 2 groups. In patients with ACS, the restenosis rate was less (P <.04) and there was a trend for target vessel revascularization to be less in covered stents (P =.09). The event-free survival rate at 4 years was 85% in the AVGCS group versus 81% in the control group (P = not significant); in ACS it was 94% versus 78%, respectively (P = not significant). Stents covered by thicker venous grafts were associated with improved clinical outcome. CONCLUSIONS: Stents covered by autologous venous grafts may be safely prepared without complications. This technique may prove to be a useful means, especially in patients with ACS. PMID- 10689259 TI - Association between total homocyst(e)ine and the likelihood for a history of acute myocardial infarction by race and ethnicity: Results from the Third National Health and Nutrition Examination Survey. AB - BACKGROUND: Few studies examining the association between total homocyst(e)ine and coronary heart disease have included blacks or Hispanics. METHODS: Data from the third National Health and Nutrition Examination Survey (3173 patients), a nationally representative survey of US adults, were used to examine the relation between total homocyst(e)ine and an electrocardiogram or a physician's diagnosis of acute myocardial infarction (259 patients) among whites, blacks, and Mexican Americans >/=40 years old. RESULTS: Vitamin B(12) and serum folate concentrations were significantly lower among persons with a total homocyst(e)ine concentration >/=15 micromol/L than among those with a total homocyst(e)ine concentration /=15 micromol/L were also older and more likely to be hypertensive, have a higher cholesterol concentration, and smoke. Compared with persons with a total homocyst(e)ine concentration /=15 micromol/L had an odds ratio (OR) for myocardial infarction of 1.8 (95% confidence interval [CI], 1.2-2.9) after adjustment for cardiovascular disease risk factors. Similar associations were noted among whites (OR 1.8, 95% CI, 1.1-3.1) and blacks (OR 1.9, 95% CI, 0.8-4.2); a more modest association was noted among Mexican Americans (OR 1.2, 95% CI, 0.3-5.0). The association between total homocyst(e)ine and myocardial infarction was also more pronounced in persons without hypertension or diabetes. CONCLUSIONS: Almost a 2-fold increased likelihood of myocardial infarction among persons with a total homocyst(e)ine concentration >/=15 micromol/L was noted in this nationally representative survey. The magnitude of the association did not differ by race or ethnicity. PMID- 10689260 TI - Cigarette smoking status and outcome among patients with acute coronary syndromes without persistent ST-segment elevation: effect of inhibition of platelet glycoprotein IIb/IIIa with eptifibatide. The PURSUIT trial investigators. AB - BACKGROUND: Studies have shown that cigarette smokers constitute a substantial proportion of patients with acute coronary syndromes (ACS) and have platelet-rich coronary thrombi. We characterized the influence of smoking status on outcome of patients with ACS without persistent ST-segment elevation and tested the hypothesis that selective inhibition of the platelet glycoprotein IIb/IIIa receptor with eptifibatide would improve outcomes among cigarette smokers. METHODS: The study population included patients enrolled in the PURSUIT trial (Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrilin Therapy) with known smoking status presenting with ischemic chest pain /=50% diameter stenosis at follow-up angiography) and clinical (target-vessel revascularization), after successful PTCA. Exercise capacity was defined as the cumulative work performed divided by body weight (watt x minutes x kilograms( 1)). RESULTS: Exercise capacity increased 43% (P <.0001) from before PTCA to 2 weeks after PTCA (early increase) and decreased 4% (P =.01) from 2 to 20 weeks after PTCA (late decrease). The gain in minimal luminal diameter (Minimal luminal diameter after - Minimal luminal diameter before) was 0.92 +/- 0.46 mm. The loss in minimal luminal diameter (Minimal luminal diameter after PTCA - Minimal luminal diameter at follow-up examination) was 0.27 +/- 0.42 mm. Exercise capacity and minimal luminal diameter measured before PTCA were positively correlated (coefficient 3.3; R = 0.12; P =.01). Gain in minimal luminal diameter correlated with the early increase in exercise capacity (coefficient -3.8; R = 0.23; P <.0001). Loss in minimal luminal diameter correlated with the late decrease in exercise capacity (coefficient 3.3; R = 0.20; P <.0001). Multivariate logistic regression analysis revealed that the late decrease in exercise capacity was independently predictive of both angiographically (odds ratio 1.13; P <.0001) and clinically (odds ratio 1.12; P <.0001) defined restenosis. CONCLUSIONS: The results demonstrated a linear relation between the severity of coronary stenosis and exercise capacity measured before PTCA. The degree of coronary luminal enlargement achieved with angioplasty and the luminal reduction that occurred between PTCA and follow-up evaluation correlated with increases and decreases in exercise capacity. Attenuation in exercise capacity was found to be a strong predictor of restenosis. PMID- 10689262 TI - Predictors of death and reinfarction at 30 days after primary angioplasty: the GUSTO IIb and RAPPORT trials. AB - BACKGROUND: Thirty-day death among recipients of fibrinolytic therapy for acute myocardial infarction (MI) is tightly correlated with easily obtainable key demographic and clinical parameters such as age, blood pressure, heart rate, and infarct location. Similar data for primary angioplasty are not available. METHODS AND RESULTS: Data from 2 large, contemporary, primary angioplasty trials were formally combined and analyzed with respect to death and death/repeat MI at 30 days through the use of multivariate logistic regression models. The 1048 patients had a median age of 62 years, and 26% were women. Thirty-eight percent had an anterior infarction. The patients underwent angioplasty at a median delay from symptom onset of 3.8 hours. Death was independently predicted by increasing age (adjusted odds ratio [OR] per decade 2.32, 95% confidence interval [CI] 1.60 to 3.42), whereas a history of smoking (OR 0.29, CI 0.13 to 0.64), Thrombolysis in Myocardial Infarction (TIMI) flow grade 3 after angioplasty (OR vs TIMI <3 0.21, CI 0.10 to 0.45) and higher systolic blood pressure (OR per 10 mm Hg 0.73, CI 0.62 to 0. 87) were associated with lower mortality rates. Death or repeat MI was independently associated with increasing age (OR per decade 1.40, CI 1.13 to 1.76) and anterior location of the index MI (OR 1.89, CI 1.12 to 3.20). TIMI grade 3 flow (OR vs TIMI <3 0.40, CI 0.23 to 0. 68) and higher systolic blood pressure (OR per 10 mm Hg 0.79, CI 0. 71 to 0.89) were associated with a lower incidence of death/repeat MI. Time to angioplasty, heart rate, extent of coronary artery disease, participation in 1 of the 2 trials, and all common coronary risk factors did not significantly predict outcome. CONCLUSIONS: Death and reinfarction after primary angioplasty are predominantly predicted by age, hemodynamic instability, and the attainment of TIMI 3 flow in the infarct artery. PMID- 10689264 TI - Differences between patients with heart failure treated by cardiologists, internists, family physicians, and other physicians: analysis of a large, statewide database. AB - BACKGROUND: The management of heart failure (HF) by cardiologists may be better than that of other physicians in that cardiologists' treatment choices more frequently conform with published guidelines and the results of clinical trials. Whether cardiologists' management of HF is more or less cost-effective is up for debate. METHODS: Information on all 1995 New York state hospital discharges assigned ICD-9-CM codes indicative of HF in the principal diagnosis position was obtained. Demographic and clinical characteristics, process of care, resource utilization, and short-term HF-related outcomes were compared between patients of cardiologists and patients of other physicians. RESULTS: A total of 44,926 patients were identified, with 10,506 (23%) receiving care from cardiologists, 28,300 (63%) from internists, 4812 (11%) from family practitioners, and 1308 (3%) from other physicians. Patients of cardiologists were younger, more frequently male, and less frequently residents of nursing homes. They were more likely to have associated cardiovascular diagnoses but less likely to have comorbid general medical conditions. Patients of cardiologists were more likely to undergo cardiac catheterization (9%) than those of internists (3%) and family practice (2%) physicians but had similar adjusted hospital length of stay and charges. Mortality and hospital readmission rates for HF were similar among the groups. Patients in the "other" group (managed mostly by surgeons) were the youngest, underwent more invasive and cardiac surgical procedures, and had the longest length of stay and highest hospital charges. CONCLUSIONS: Cardiologists' management of HF is not economically disadvantageous. The relations among physician specialty, process of care, resource utilization, and clinical outcomes require further study before rational and evidence-based health care staffing recommendations can be formulated. PMID- 10689265 TI - Detection of left ventricular regional relaxation abnormalities and asynchrony in patients with hypertrophic cardiomyopathy with the use of tissue Doppler imaging. AB - BACKGROUND: It is well known that the distribution and magnitude of left ventricular (LV) hypertrophy are not uniform in patients with hypertrophic cardiomyopathy (HCM), which results in regional heterogeneity of LV early diastolic function. The advent of tissue Doppler imaging (TDI) has allowed the noninvasive evaluation of regional LV wall motion velocities. The aim of this study was to evaluate regional LV relaxation abnormalities and asynchrony noninvasively in patients with HCM by using pulsed and color-coded TDI. METHODS AND RESULTS: We studied 20 patients with asymmetric septal hypertrophy (HCM group) and 18 age-matched normal patients (control group). The peak early diastolic motion velocity (Ew) and time from the aortic component of the second heart sound to the peak of the Ew (II(A)-Ew) were measured by pulsed TDI. The myocardial velocity gradient during early diastole (MVG-Ew) also was measured by color-coded TDI. Mean values for these parameters were determined on the basis of measurements made at 2 sites of the ventricular septum or posterior wall at the levels of chordae tendineae and papillary muscles. The mean Ew and mean MVG-Ew for the ventricular septum and posterior wall were significantly lower, and mean II(A)-Ew was significantly prolonged in the HCM group compared with the control group. This difference was most pronounced in the hypertrophied ventricular septum of the HCM group. The standard deviations of II(A)-Ew for the ventricular septum and posterior wall were significantly greater in the HCM group than in the control group. The time constant of LV pressure decay during isovolumic diastole (tau) correlated inversely with Ew and MVG-Ew and correlated directly with II(A) Ew. Furthermore, tau correlated directly with the standard deviation of the II(A) Ew. CONCLUSIONS: LV early diastolic function in patients with HCM may be mediated by an augmentation of regional LV relaxation abnormalities and asynchrony. PMID- 10689266 TI - Effects of amlodipine on exercise tolerance, quality of life, and left ventricular function in patients with heart failure from left ventricular systolic dysfunction. AB - BACKGROUND: A preliminary study suggested that the long-acting late-generation calcium-channel blocker amlodipine has favorable effects on exercise tolerance and is safe to use in heart failure, in contrast to earlier generation agents. The goal of 2 multicenter studies was to assess the effect of adjunctive therapy with amlodipine in addition to standard therapy on exercise capacity, quality of life, left ventricular function, and safety parameters in patients with heart failure and left ventricular systolic dysfunction. METHODS: Two large multicenter trials examining the effects of amlodipine on these parameters over a 12-week period of therapy were undertaken in patients with mild to moderate heart failure and left ventricular systolic dysfunction. A total of 437 patients with stable heart failure were studied in a randomized, double-blind, placebo-controlled prospective design. RESULTS: Amlodipine at a dose of 10 mg/day in addition to standard therapy in such patients was associated with no significant difference in change in exercise tolerance on a Naughton protocol compared with placebo in each trial. Among all patients taking amlodipine, exercise time increased 53 +/- 9 (SE) seconds; exercise time for those taking placebo increased 66 +/- 9 seconds (P = not significant). There were no significant differences in changes of quality of life parameters between amlodipine- and placebo-treated patients, and there were no significant differences in symptom scores or New York Heart Association classification between groups. Left ventricular function (measured as ejection fraction) improved 3. 4% +/- 0.5% in amlodipine-treated patients and 1.5% +/- 0.5% in placebo-treated patients (P =.007). There was no statistically significant excess of important adverse events (episodes of worsening heart failure in 10% amlodipine-treated vs 6.3% of placebo-treated patients) or differences in need for changes in background medication between groups. CONCLUSIONS: The addition of 10 mg of amlodipine per day to standard therapy in patients with heart failure is associated with no significant improvement in exercise time compared with placebo therapy over a 12-week period, and there was no increased incidence of adverse events. These data suggest that the addition of amlodipine to standard therapy in heart failure will not result in additional efficacy per se beyond standard therapy. PMID- 10689267 TI - A rationale for the use of beta-blockers as standard treatment for heart failure. AB - BACKGROUND: Cardiac sympathetic activation is one of the major and earlier changes observed in patients with heart failure. Its relation to the severity of the disease and its independent prognostic value show that it may directly contribute to the progression of heart failure. beta-Blockers are the most effective tool to counteract the untoward effects of sympathetic activation on the cardiovascular system. METHODS AND RESULTS: We reviewed the results of the placebo-controlled, double-blind studies about the effects of beta-blockers in patients with heart failure. These studies have involved almost 10,000 patients to date and have consistently shown that the long-term administration of beta blockers is associated with a highly significant improvement in both left ventricular function and prognosis of the patients with heart failure. The evidence supporting the use of beta-blockers now equals or even surpasses that of angiotensin-converting enzyme inhibitors; therefore beta-blockers should be considered part of standard therapy. Issues that remain unclarified include the mechanisms through which beta-blockers may improve cardiac function and their tolerability and efficacy in specific groups of patients (such as those with asymptomatic left ventricular dysfunction, severe heart failure, the elderly, or those with left ventricular diastolic dysfunction). It is not currently clear whether the pharmacologic differences between individual beta-blockers are clinically relevant. If they are, the potential for even greater benefit with certain agents exists. It is hoped that these issues will be clarified by the results of ongoing multicenter trials. PMID- 10689268 TI - Magnesium supplementation in the prevention of arrhythmias in pediatric patients undergoing surgery for congenital heart defects. AB - BACKGROUND: The efficacy of magnesium in the prevention of arrhythmias in pediatric patients after heart surgery remains unknown. Therefore we prospectively examined the effect of magnesium treatment on the incidence of postoperative arrhythmias in pediatric patients undergoing surgical repair of congenital heart defects. METHODS AND RESULTS: Twenty-eight pediatric patients undergoing heart surgery with cardiopulmonary bypass were prospectively, randomly assigned in a double-blind fashion to receive intravenous magnesium (magnesium group, n = 13; 30 mg/kg) or saline (placebo group, n = 15) immediately after cessation of cardiopulmonary bypass. Magnesium, potassium, and calcium levels were measured at defined intervals during surgery and 24 hours after surgery. Continuous electrocardiographic documentation by Holter monitor was performed for 24 hours after surgery. Magnesium levels were significantly decreased below the normal reference range for patients in the placebo group compared with the magnesium group on arrival in the intensive care unit and for 20 hours after surgery. Magnesium levels remained in the normal range for patients in the magnesium group after magnesium supplementation. In 4 patients in the placebo group (27%), junctional ectopic tachycardia developed within the initial 20 hours in the intensive care unit. No junctional ectopic tachycardia was observed in the magnesium group (P =.026). CONCLUSIONS: Although this study was originally targeted to include 100 patients, the protocol was terminated because of the unacceptable incidence of hemodynamically significant junctional ectopic tachycardia that was present in the placebo group. Thus low magnesium levels in pediatric patients undergoing heart surgery are associated with an increased incidence of junctional ectopic tachycardia in the immediate postoperative period. PMID- 10689269 TI - Left ventricular function, cardiac dysrhythmias, atrial activation, and volumes in nondipper hypertensive individuals with left ventricular hypertrophy. AB - BACKGROUND: Arrhythmic patterns and left ventricular geometric adaptations to pressure overload were investigated in 76 patients with untreated borderline-to moderate sustained essential hypertension studied by 2-dimensional and M-mode echocardiography, 12-lead, Holter, and signal-averaged electrocardiography, and ambulatory blood pressure monitoring. METHODS AND RESULTS: Sixty-two age- and sex matched normal adults were chosen for data comparison. Hypertrophic hypertensive patients were subdivided into 2 subgroups: 44 patients with nocturnal blood pressure reduction (dippers) and 32 patients without it (nondippers). Common afterload and diastolic function indexes were found to be lower in combined nondipper and dipper groups, but only fractional shortening decreased in nondippers. The number of premature atrial and ventricular contractions per hour was high in dippers and nondippers, with no statistically significant differences between them; atrial and ventricular complex dysrhythmias were similar. Signal averaged electrocardiography showed a prolonged P-wave duration in dipper and nondipper patients with high atrial volumes but no late ventricular potentials and no difference in quantitative P-wave analysis. Left atrial volumes, P-wave duration, and premature atrial contractions were found to be positively linked to left ventricular hypertrophy. In nondipper patients a linear correlation was observed between left atrial volume and P-wave duration, although supraventricular ectopic activity was connected to left atrial volume enlargement both in dipper and nondipper patients. CONCLUSIONS: These data suggest that the nondipper pattern is not linked to a worse arrhythmogenic substrate; only atrial volume increase may be related to significant supraventricular activity and prolonged atrial activation in nondipper patients, but late ventricular potentials are uncommon in hypertrophic hypertensive patients. PMID- 10689270 TI - beta2-adrenergic receptor polymorphisms at amino acid 16 differentially influence agonist-stimulated blood pressure and peripheral blood flow in normal individuals. AB - BACKGROUND: The Gly16 beta(2)-adrenergic receptor (beta(2)AR) polymorphism is a common variant of the beta(2)AR that displays depressed function caused by enhanced receptor downregulation in vitro compared with the Arg16 receptor. METHODS AND RESULTS: We studied 20 healthy, normotensive, nonsmoking white individuals who were homozygous for either the Arg16 (n = 10) or the Gly16 (n = 10) genotype. Plethysmographic lower-limb blood flow, blood pressure, and 2 dimensional echocardiograms were recorded at baseline and after 15-minute incremental infusions of terbutaline (100 to 300 ng/kg per minute). Baseline heart rates, blood pressures, and flows were similar in both groups, but at the maximum dose of terbutaline, limb blood flow was less (P <.05), calculated vascular resistance was greater (P <.05), and systolic and diastolic blood pressures were greater in patients with Gly16 than in those with Arg16 (both P <.05). In contrast, terbutaline-stimulated heart rates were not different. In a separate group of 20 homozygous individuals (12 Arg16, 8 Gly16), there were no differences in 2-dimensional echocardiographically determined ventricular function. CONCLUSIONS: We conclude that the Gly16 beta(2)AR polymorphism imparts attenuated vasodilatory responses to catecholamines in normal human beings and is an important genetic component in the regulation of peripheral blood flow and systemic arterial pressure. PMID- 10689271 TI - A controlled trial of cardiac rehabilitation in the home setting using electrocardiographic and voice transtelephonic monitoring. AB - OBJECTIVE: The goal of this study was to compare the effectiveness of home-based, transtelephonically monitored cardiac rehabilitation with standard, on-site, supervised cardiac rehabilitation. BACKGROUND: Participation in cardiac rehabilitation has been demonstrated to increase exercise capacity, decrease cardiovascular symptoms, improve psychosocial status, and decrease total and cardiovascular mortality rates in patients with coronary heart disease. Because of multiple factors, national overall participation is only at 15% of eligible patients. METHODS: Effects of a 3-month home-based, transtelephonically monitored rehabilitation program (n = 83 patients) with simultaneous voice and electrocardiographic transmission to a centrally located nurse coordinator were compared with effects of a standard on-site rehabilitation program (n = 50 patients). The study design was a multicenter, controlled trial. Primary outcome variables were peak aerobic capacity and quality of life, as measured by the Health Status Questionnaire. RESULTS: Patients in the home-based monitoring program increased peak aerobic capacity to a similar degree as patients who exercised on site (18% vs 23%). Quality of life domains of physical functioning, social functioning, physical role limitations, emotional role limitations, bodily pain, and energy/fatigue improved similarly in both groups. There were no circulatory arrests or other major exercise-related medical events in either group. A total of 3100 hours of home exercise were transtelephonically monitored. CONCLUSIONS: Patients with coronary heart disease can effectively participate in home-based, monitored cardiac rehabilitation, with exercise and quality of life improvements comparable to those demonstrated at on-site programs. PMID- 10689272 TI - Mitral repair in patients with a ruptured papillary muscle. AB - BACKGROUND: The objective of this study was to evaluate the feasibility of a modified papillary muscle repair procedure for a group of patients with ischemic mitral regurgitation when ischemia/infarction has resulted in the rupture of a papillary muscle. From January 1997 to January 1999, 843 patients underwent mitral valve surgery in our hospital. Mitral reconstruction was performed in 520 (61.7%) patients, and 6 (1.2%) of these patients were found to have a rupture of a papillary muscle at initial examination. METHODS AND RESULTS: A modified papillary muscle repair procedure to reimplant the tip of the ruptured papillary muscle "height- and/or length-adjusted" into a corresponding papillary muscle, with the use of a sandwiched pericardium pledget-reinforced polytetrafluoroethylene suture, was performed in 6 patients. Although the underlying cause in this group of patients was ischemic, concomitant coronary artery bypass grafting was performed in only 3 patients, with 1.3 grafts per patient. Of these 6 patients, 3 (50%) were men; the mean age was 60.2 +/- 12.8 years. All patients had in addition to the papillary muscle repair procedure an annuloplasty with a Carpentier-Edwards Physio-Ring. There was no early death in this group of patients. Postoperative Doppler echocardiography showed satisfactory mitral valve function in all patients and a significant postoperative ventricular remodeling: The left ventricular end-diastolic diameter decreased from 72.8 +/- 3.1 mm before surgery to 54.6 +/- 9.3 mm (P <.1) after surgery; left ventricular systolic diameter also decreased (48.5 +/- 4.9 mm vs 38. 4 +/- 9.8 mm; P <.1), and a substantial reduction of left atrial diameter (58.8 +/- 1.5 mm vs 49.7 +/- 4.1 mm; P <.1) was observed. Within the short mean follow-up period of 8.6 +/- 7.5 months (2 to 26 months), there were no late deaths, reoperations, or thromboembolic or bleeding complications. All patients were in New York Heart Association functional class I or II at the time of follow up. CONCLUSIONS: Our results indicate that our modified papillary muscle reimplantation procedure is a valuable surgical tool with good survival results in patients with ischemic mitral regurgitation caused by papillary muscle rupture. PMID- 10689273 TI - Lipoprotein(a) in atrial fibrillation. PMID- 10689275 TI - Effects of exercise on QT dispersion in ischemic heart disease. PMID- 10689277 TI - Low-molecular-weight heparin as optimal solution to therapeutic heparinization. PMID- 10689279 TI - Increased expression of hemostasis markers in the coronary circulation in patients with coronary artery disease. PMID- 10689280 TI - Comparison of the effects of the mother and an unfamiliar adult female on cortisol and behavioral responses of pre- and postweaning guinea pigs. AB - In the guinea pig, the presence of the mother, but not littermates, has been found to inhibit hypothalamic-pituitary-adrenal (HPA) responses during brief (30 60 min) exposure to novel surroundings both prior to and several weeks following the completion of weaning. In the present study, we found that an unfamiliar adult female inhibited plasma cortisol and vocalization responses of pre- and postweaning guinea pigs during a 60-min exposure to a novel environment. However, the presence of the mother still had a greater effect on the cortisol levels of the young, at least during the preweaning period. The moderating influence of the unfamiliar adult female on vocalizations and cortisol levels occurred despite behavioral interactions, such as heightened aggression and sexual behavior, not seen during tests with the mother. It is suggested that the unfamiliar adult female's effectiveness in reducing HPA activity during exposure to novelty may facilitate the change in patterns of social interaction occurring in recently weaned animals. PMID- 10689281 TI - The effect of early experience on learning and memory in cuttlefish. AB - The effect of early experience on the growth and ontogeny of memory in cuttlefish (Sepia officinalis) was studied using an associative learning protocol. Five groups of cuttlefish were reared in different conditions (standard conditions, SC; impoverished conditions, IC; enriched conditions, EC; impoverished then enriched conditions, I/EC; enriched then impoverished conditions, E/IC) from birth to the 3rd month of postembryonic life. Acquisition and retention of the learning task were assessed at 1 and 3 months. Growth was slower and maturation of memory abilities occurred later in cuttlefish from Group IC than in cuttlefish from Group EC, with the maturation rate of memory in cuttlefish from Group SC intermediate between these two groups. Retention performances of cuttlefish from Groups I/EC and E/IC indicated that the environment of rearing during the 2nd and/or 3rd months of life was crucial for the development of memory. PMID- 10689282 TI - Spontaneous kicking behavior in infants: age-related effects of unilateral weighting. AB - The age-related effects of unilateral weighting on spontaneously generated kicks in 18 healthy, full-term infants were investigated. The main question was whether infants during the first half-year after birth reveal changes in how they adjust to unilateral weighting. At 6 weeks, infants reduced the frequency of the weighted leg and increased that of the unweighted leg whereas at 12 weeks the frequency of kicking increased in both legs. At both ages, unilateral weighting also resulted in differences on a number of kinematic parameters. By 18 and 26 weeks, such frequency and kinematic effects were no longer present. With regard to interlimb couplings, a clear pattern of bilateral coordination was only present at 26 weeks; these results suggest that the effects of unilateral weighting are not directly related to the tightness of interlimb couplings. The implications of these age-related differences for understanding developmental changes in the control of leg movements are discussed. It is suggested that the infants' improved ability to act in a task-specific manner as well as nonlinear changes in the musculo-skeletal system and fine-tuning processes at a neural level might be factors of importance. PMID- 10689283 TI - A dissociation in infants' memory for stimulus size: evidence for the early development of multiple memory systems. AB - Adults' memory performance on recognition (explicit memory) tests is sensitive to stimulus size, but their performance on priming (implicit memory) tests is not. This memory dissociation is taken as evidence for two, functionally distinct memory systems. Young infants, however, are thought to possess only a single representational system that supports implicit memory; the system that supports explicit memory is thought not to mature before 8-9 months of age. In two experiments with 54 infants, we asked if 3-month-olds exhibit a memory dissociation for stimulus size on recognition and priming tests. All infants learned to move a mobile displaying +s of a given size. In Experiment 1, infants recognized +s in the original size but not 33% smaller or larger. In Experiment 2, +s were effective memory primes in a reactivation task, irrespective of size. The finding that young infants exhibit a memory dissociation for stimulus size adds to growing evidence that two memory systems are functional from early in development. PMID- 10689284 TI - Comparative analyses of the role of postnatal development on the expression of play fighting. AB - Whether it is that animals are young so that they can play, or whether it is that they play because they are young, play should be more prevalent in species that have a greater degree of postnatal development. This hypothesis is tested by comparative analyses within two mammalian orders (primates and muroid rodents) using independent contrasts. This technique can account for the relative degree of relatedness among the species. For both orders, the complexity or prevalence of play fighting is compared to the degree of prenatal development (neonatal weight/adult weight). In addition, the prevalence of play in primates is compared to prenatal brain development (neonatal brain weight/adult brain weight). Significant negative regressions show that 30% of the variance in the distribution of play in the rodents is accounted for by the degree of prenatal development of body size, and 60% of the variance in play in the primates is accounted for by prenatal brain growth. The findings are thus consistent with the prediction. Species with a greater proportion of their growth occurring postnatally play more and have more complex play than do species with more of their growth occurring prenatally. PMID- 10689285 TI - Ontogeny of delay versus trace eyeblink conditioning in the rat. AB - The ontogeny of delay versus trace eyeblink conditioning was examined in 19-, 23 , and 30-day-old rat pups. Pairings of a tone conditioned stimulus (CS) and periocular shock unconditioned stimulus (US; 100-ms) were presented in one of three conditioning paradigms: standard delay [380-ms CS, 280-ms interstimulus interval (ISI)], trace (380-ms CS, 500-ms trace interval), or long-delay (980-ms CS, 880-ms ISI). The results of two experiments indicated that standard delay conditioning emerged between 19 and 23 days of age whereas trace and long-delay eyeblink conditioning emerged more slowly from postnatal Days 19 to 30. Because the acquisition profile for long-delay paralleled that of trace and not standard delay, it appears that the relative deficits in the emergence of trace eyeblink conditioning during development reflect difficulty in forming associations over long ISIs rather than the short-term memory demands of the trace conditioning paradigm. PMID- 10689286 TI - The development of inhibitory control in preschool children: effects of "executive skills" training. AB - As one of several processes involved in the executive functioning of the cognitive system, inhibitory control plays a significant role in determining how various mental processes work together in the successful performance of a task. Studies of response inhibition have shown that although 3-year-old children have the cognitive capacity to learn the rules required for response control, indicated by the correct verbal response, developmental constraints prevent them from withholding the correct response (Bell & Livesey, 1985; Livesey & Morgan, 1991). Some argue that these abulic dissociations are relative to children's ability to reflect on the rules required for response control (Zelazo, Reznick, & Pinon, 1995). The current study showed that repeated exposure to tasks facilitating the acquisition of increasingly complex rule structures could improve inhibitory control (as measured by a go/no-go discrimination learning task), even in children aged 3 years. These tasks included a variant of Diamond and Boyer's (1989) modified version of the Wisconsin Card Sort Task and a simplification of the change paradigm (Logan & Burkell, 1986). It is argued that experience with these tasks increased the acquisition of complex rules by placing demands on executive processes. This includes response control and other executive functions, such as representational flexibility, the ability to maintain information in working memory, the selective control of attention, and proficiency at error correction. The role of experiential variables in the development of inhibitory control is discussed in terms of the interaction between neural development and appropriate executive task experience in the early years. PMID- 10689287 TI - Facile entry to (-)-(R)- and (+)-(S)-mexiletine. AB - The title compounds, 1a and 1b, have been synthesized in a three-step sequence starting from (-)-(S) and (+)-(R)-propylene oxide, respectively, in acceptable overall yields. The enantiomeric excess values for 1a and 1b were 96% and 93% respectively, as assessed by HPLC analysis on a chiral stationary phase of the corresponding N-acetyl derivatives. The synthetic route herein presented may represent a facile entry to highly enriched mexiletine enantiomers, alternative to those previously reported in the literature. PMID- 10689288 TI - Mechanisms of retention of pyrrolidinyl norephedrine on immobilized alpha(1)-acid glycoprotein. AB - The HPLC separation of the R,S and S,R enantiomers of pyrrolidinyl norephedrine on immobilized alpha-1 glycoprotein (AGP) was investigated. Conditions for the separation were varied using a premixed mobile phase containing an ammonium phosphate buffer and an organic modifier. The influence of mobile phase pH, ionic strength, organic modifier composition, modifier type, and temperature on the chiral selectivity and retention were investigated. The presented data demonstrate that independent phenomena govern the enantioselectivity and retention. Retention is a function of both ion exchange equilibria and hydrophobic adsorption. Thermodynamic data derived from van't Hoff plots illustrates that while enantioselectivity is also enthalpically driven, the magnitude of the enthalpy term is governed by pH. Enantioselectivity has little dependence on ionic strength. Hydrophobic interactions appear to foster hydrogen bonding interactions; the two appear to be mutually responsible for chiral selectivity. The chiral selectivity decreases as the pH is decreased and increases with mobile phase buffer strength. PMID- 10689289 TI - Parity violation and the evolution of biomolecular homochirality. AB - Parity violation at the level of terrestrial biopolymers, as seen in proteins, DNAs, and RNAs, and parity violation at the level of nuclear processes, as evident in longitudinally polarized beta-particles and parity-violating energy differences (PVEDs), are discussed and their fundamental importances are emphasized. Attempts to find a causal connection between the unique homochirality of biopolymers and parity violation at the nuclear level, and speculations that the former is a consequence of the latter, are reviewed. Consideration of all lines of evidence leads to the conclusion that there is no substantiation for such a causal connection, and that the two levels of parity violation are entirely independent of each other. PMID- 10689290 TI - Base-catalyzed epimerization of the butenolide in annonaceous acetogenins. AB - The elusive epimerization process in the chiral butenolide moiety of Annonaceous acetogenins was examined under several sets of conditions commonly used for elimination leading to the alpha, beta-unsaturated lactone and the results provide practical guidance in choosing elimination conditions. PMID- 10689291 TI - On the calculation of Gibbs energy corresponding to enantioselective interactions at a direct HRGC separation of enantiomers. AB - A novel procedure is proposed for the calculation of Gibbs energy corresponding to enantiospecific interactions of 2-(2, 4-dinitrophenoxy)-, 2-phenoxy-, and 2 halogen-n-pentane enantiomers with a beta-cyclodextrin (ChirasilDex) stationary phase under gas chromatographic conditions. This energy is calculated from retention data as a difference between the Gibbs energy of an enantiomer and its corresponding achiral congener. The procedure for the determination of 2-(2,4 dinitrophenoxy)-, 2-phenoxy- and 2-halogen- n-pentane achiral congener retention data is discussed in detail. PMID- 10689292 TI - Determination of the absolute configuration of (-)-(3R)-O-beta-D glucopyranosyloxy-5-phenylpentanoic acid from Polygonum salicifolium. AB - The absolute configuration of the title compound has been determined after its enzymatic hydrolysis to 3-hydroxy-5-phenylpentanoic acid, esterification, and identification of the enantiomerically pure methyl (3R)-hydroxy-5 phenylpentanoate by HPLC on Chiralcel(R)OD-H. For reasons of inconsistent literature data, enantioselective reductions of methyl 3-oxo-5-phenylpentanoate have been reinspected and the stereochemical outcome unequivocally confirmed by both chiroptical and HPLC retention data. PMID- 10689293 TI - Synthesis of both enantiomers of 4-vinyl oxazolidin-2-one from a single precursor: D-isoascorbic acid. AB - A convenient synthesis of (S)- and (R)-4-vinyl oxazolidin-2-one 1 and 2 from the same inexpensive starting material, D-isoascorbic acid, is described. The title compounds were obtained in 44% and 38% yield, respectively, by operationally simple steps. This approach is a suitable alternative to the literature methods and enhances the synthetic utility of these intermediates. Inc. PMID- 10689294 TI - Chiral photogeneration using perfluorocyclopentene-derived photochromes. AB - A new type of perfluorocyclopentenes which contain an optically active group at the 2-position of the thiophene ring were synthesized. Irradiation with UV light afforded the cyclized diastereoisomers in ratios dependent on solvent polarity and temperature. PMID- 10689295 TI - Nefopam enantiomers: preclinical pharmacology/toxicology and pharmacokinetic characteristics in healthy subjects after intravenous administration. AB - Nefopam (NEF) is a potent analgesic compound administered as a racemic mixture. Previous in vitro and in vivo studies with nefopam enantiomers have shown that (+)nefopam [(+)NEF] is substantially more potent than (-)nefopam [(-)NEF]. Differences between enantiomers have also been suggested in metabolic studies in vitro. The impact of these differences in vivo is not known because there is little or no information on the relative plasma concentrations of the enantiomers or on their kinetics. In this study, individual enantiomers of nefopam were synthesized and examined for acute toxicity in male and female rats and mice. Pharmacologic properties of enantiomers were examined using in vitro binding assays and antinociceptive tests in rats and mice. Additionally, a pharmacokinetic study was conducted in human volunteers. Subjects were administered 20 mg nefopam as Acupan(R) either as a 5- or 20-min intravenous infusion. In a control phase, subjects were administered only vehicle. Blood samples were collected through the following 24 h. Plasma samples were analyzed for individual enantiomers using a chiral assay developed for this purpose. The pharmacologic differences of previous studies were confirmed in receptor binding assays and in the hot plate and the formalin tests in mice. Neither enantiomer demonstrated substantial activity in the tail flick test in rats. No significant differences were revealed between LD(50) values of nefopam enantiomers after oral or intravenous administration in male and female rats or mice. There were no significant differences in AUC(0-infinity), C(max), or half-life between enantiomers following intravenous administration. Based on these findings, there is currently no compelling rationale to justify administering or monitoring individual enantiomers. PMID- 10689296 TI - Tumor necrosis factor: a master-regulator of leukocyte movement. PMID- 10689297 TI - The emerging role of IL-15 in NK-cell development. PMID- 10689298 TI - Microchimerism in human diseases. PMID- 10689299 TI - Antigen processing and presentation by intestinal epithelial cells - polarity and complexity. AB - The mechanisms by which gut-associated lymphoid tissue (GALT) maintains a balance between oral tolerance and active immune response in the face of exposure to high antigen concentrations remains a central question in mucosal immunity. Here, Robert Hershberg and colleagues discuss the evidence that human intestinal epithelial cells function as antigen-presenting cells (APCs) capable of regulating T-cell responses in the intestinal mucosa PMID- 10689300 TI - Heat shock proteins, tumor immunogenicity and antigen presentation: an integrated view. AB - A broad range of studies has established that heat shock proteins (Hsps) potentially play a role in tumor immunosurveillance. Here, Andrew Wells and Miroslav Malkovsky highlight recent data that demonstrate a causal relationship between the expression of Hsps and tumor immunogenicity, and suggest several mechanisms by which Hsps might influence the capacity of a tumor to induce an immune response. PMID- 10689301 TI - The brain and thymus have much in common: a functional analysis of their microenvironments. AB - Research into the neural and immune systems has begun to converge. Since the first reports that interleukins play important roles in both systems and that lymphocytes secrete neuronal factors, scientists have been surprised by the ever increasing list of interactions. Here, Rolf Mentlein and Marion Kendall examine the major supporting cells of the brain and thymus - astrocytes and thymic epithelial cells - the similar neuroectodermal origin of which could explain such fundamental analogies. PMID- 10689302 TI - Regulation of T-cell responses by CNS antigen-presenting cells: different roles for microglia and astrocytes. AB - Analysis of the mechanisms underlying CNS immune surveillance and immunopathology have provided new insights into the intracerebral regulation of immune responses. Here, Francesca Aloisi, Francesco Ria and Luciano Adorini review the role of CNS antigen presenting cells and focus on the control of Th1 and Th2 responses by microglia and astrocytes. PMID- 10689303 TI - Tyrosine kinase SYK: essential functions for immunoreceptor signalling. AB - The tyrosine kinase SYK plays critical roles in signalling through immune receptors. Gene-targeting studies have identified the cell types that require SYK for development and function, and the receptors that use SYK as well as their downstream signalling effectors. There is also evidence of a role for SYK in non immune cells and in the maintenance of vascular integrity. PMID- 10689304 TI - Counting antigens using antibodies. PMID- 10689305 TI - Allergenicity of grass and oil seed rape pollen. PMID- 10689306 TI - Reply to jones et al PMID- 10689307 TI - Gene therapy: a rocky start to the new millennium PMID- 10689308 TI - New predictive test for high blood pressure. PMID- 10689310 TI - New NIDDK head predicts greater emphasis on genetics. PMID- 10689309 TI - Ambitious plans to make sense of signalling. PMID- 10689311 TI - Australia faces biomedical brain drain. PMID- 10689313 TI - How does HIV cause AIDS? The homing theory. AB - The mechanism by which HIV causes depletion of CD4+ T cells in infected individuals remains unknown. Numerous theories have been proposed, but none can fully explain all of the events observed to occur in patients. Recent studies have shown that HIV binding to resting CD4+ T cells upregulates L-selectin, causing the cells to home from the blood into lymph nodes at an enhanced rate. It is possible that the disappearance of CD4+ T cells in the blood is actually the result of them leaving the blood, which can help explain the loss of CD4+ T cells in the blood occurring at a much faster rate than in lymphoid tissues. Furthermore, secondary signals through homing receptors received during the homing process induce many of these cells into apoptosis. These cells die in the lymph nodes without producing HIV particles, which can explain the 'bystander effect' observed in the lymph nodes of HIV infected individuals. If this scenario occurs in HIV+ patients, it might explain many of the clinical observations. PMID- 10689314 TI - Knowing left from right: the molecular basis of laterality defects. AB - The apparent symmetry of the vertebrate body conceals profound asymmetries in the development and placement of internal organs. Asymmetric organ development is controlled in part by genes expressed asymmetrically in the early embryo, and alterations in the activities of these genes can result in severe defects during organogenesis. Recently, data from different vertebrates have allowed researchers to put forward a model of genetic interactions that explains how asymmetric patterns of gene expression in the early embryo are translated into spatial patterns of asymmetric organ development. This model helps us to understand the molecular basis of a number of congenital malformations in humans. PMID- 10689315 TI - Synthetic surfactants to treat neonatal lung disease. AB - Pulmonary surfactant is a complex of surface-active lipids mixed with specific proteins. Two of these, SP-B and SP-C, are essential for adsorption of surfactant lipids to the air-liquid interfaces of the lungs and, hence, are also essential for alveolar stability and effective gas exchange. Surfactant substitutes must contain at least one of these proteins (or analogues of them) to be optimally effective when administered into the airways of babies with surfactant deficiency or dysfunction. This review describes how an increased understanding of the properties of surfactant proteins has led to the development of improved synthetic surfactants with the potential to treat a wide range of respiratory disorders. PMID- 10689316 TI - Superantigens - powerful modifiers of the immune system. AB - Superantigens are powerful microbial toxins that activate the immune system by binding to class II major histocompatibility complex and T-cell receptor molecules. They cause a number of diseases characterized by fever and shock and are important virulence factors for two human commensal organisms, Staphylococcus aureus and Streptococcus pyogenes, as well as for some viruses. Their mode of action and variation around the common theme of over-stimulating T cells, provides a rich insight into the constant battle between microbes and the immune system. PMID- 10689317 TI - Animal models of stroke. PMID- 10689318 TI - Medical and veterinary malacology in Africa. PMID- 10689319 TI - Biochemistry of Plasmodium on the Web. PMID- 10689321 TI - Health Information for International Travel 1999-2000. PMID- 10689320 TI - Malaria Research and Reference Reagent Resource Center. PMID- 10689322 TI - Ascaris haemoglobin: new tricks for an old protein. PMID- 10689323 TI - Insecticide products: treatment of mosquito nets at home. PMID- 10689325 TI - Functional Websites for Parasite Genome Projects. PMID- 10689324 TI - Leishmaniasis, HIV and Oral Treatments on the Web. PMID- 10689326 TI - Plasmodium falciparum Genome Update. PMID- 10689327 TI - Do eosinophils have a role in the killing of helminth parasites? AB - Eosinophils have been shown to be potent effector cells for the killing of helminth parasites in in vitro cultures. However, an in vivo role for eosinophils has been more difficult to establish. Early data showed close associations between eosinophils and damaged or dead parasites in histological sections, and significant correlations between resistance to parasites and the capacity to induce eosinophilia after infection. However, more recent studies, using mice that have reduced or increased eosinophil levels through targeting of the eosinophil-specific cytokine interleukin 5, have not unanimously supported an in vivo role for eosinophils in resistance to parasites. Here, Els Meeusen and Adam Balic review these studies and suggest a major role for the innate immune response in unnatural mouse-parasite models to explain some of the findings. They conclude that the data so far are consistent with a role for eosinophils in the killing of infective larval stages, but not adults, of most helminth parasites. PMID- 10689328 TI - Commitment to gametocytogenesis in Plasmodium falciparum. AB - To achieve transmission, a subpopulation of asexually dividing bloodstream forms of the human malaria parasite Plasmodium falciparum withdraws from the cell cycle to develop into gametocytes - cells specialized for sexual reproduction and invasion of the mosquito vector. For natural selection to maximize transmission to new hosts, a balance must have evolved between asexual replication and sexual differentiation. Here, Mike Dyer and Karen Day consider observations on the process of commitment to gametocytogenesis and use this information as the framework for a model that begins to explain the control of the dynamics between asexual and sexual development. PMID- 10689329 TI - The kidney form of Trypanosoma musculi: A distinct stage in the life cycle? AB - Trypanosoma musculi is a parasite specific to mice, which resides in the blood and lacks intracellular stages. After immune clearance of the flagellates from the general circulation, mice are resistant to reinfection. Yet, long after parasites are no longer detected in the peripheral blood, they persist in the vasa recta of the kidneys and it has been proposed that this is an immunologically privileged site for T. musculi. This relationship provides a useful model for studies of latent or chronic infections in immune hosts. Here, Fernando Monroy and Donald Dusanic consider the immune responses of mice to T. musculi and compare characteristics of the parasites from the vasa recta (kidney forms, KFs) of mice with latent infections to trypanosomes from the peripheral blood (bloodstream forms, BSFs) of animals during active infections. They consider how KFs evade immune destruction and suggest that these sequestered parasites represent a distinct stage in the life cycle. PMID- 10689330 TI - Progress in the serodiagnosis of Neospora caninum infections of cattle. AB - Neospora caninum is an apicomplexan protozoan that has become the focus of significant research attention worldwide. This organism infects a range of host species, including dogs, from which it was originally reported in 1984, but it is most important as a major cause of bovine abortion. As a result of the global importance of N. caninum, researchers have developed a number of serological tests to investigate the epidemiology of infection and disease. In this article, Robert Atkinson, Peter Harper, Michael Reichel and John Ellis consider progress made in the serodiagnosis of N. caninum. PMID- 10689331 TI - Tsetse--A haven for microorganisms. AB - Arthropods are involved in the transmission of parasitic and viral agents that cause devastating diseases in animals and plants. Effective control strategies for many of these diseases still rely on the elimination or reduction of vector insect populations. In addition to these pathogenic organisms, arthropods are rich in microbes that are symbiotic in their associations and are often necessary for the fecundity and viability of their hosts. Because the viability of the host often depends on these obligate symbionts, and because these organisms often live in close proximity to disease-causing pathogens, they have been of interest to applied biologists as a potential means to genetically manipulate populations of pest species. As knowledge on these symbiotic associations accumulates from distantly related insect taxa, conserved mechanisms for their transmission and evolutionary histories are beginning to emerge. Here, Serap Aksoy summarizes current knowledge on the functional and evolutionary biology of the multiple symbionts harbored in the medically and agriculturally important insect group, tsetse, and their potential role in the control of trypanosomiasis. PMID- 10689332 TI - Should DDT be banned by international treaty? AB - The insecticide DDT has been an effective and affordable means of malaria control in many countries, but pressure for its use to be banned is mounting. Here, Chris Curtis and Jo Lines take a critical look at evidence that links house spraying by DDT with harm to the environment and human health, and stress the need for resources for alternatives to DDT to be made available to countries that would be affected by a DDT ban. PMID- 10689333 TI - The prophylactic effects of artemether against Schistosoma japonicum infections. AB - The fight against schistosomiasis in China has been very effective in reducing the number of infections across the country. However, the drug of choice, praziquantel, has no prophylactic effect, which reduces its efficacy in high transmission areas. This situation has prompted efforts to find prophylactic compounds, the most promising of which is the drug artemether. In this article, Xiao Shuhua, Mark Booth and Marcel Tanner review the results of laboratory tests and field trials of artemether against schistosomiasis in China. PMID- 10689334 TI - Identification of parasitic genes by computational methods. AB - A number of parasite genome projects are under way, and large amounts of nucleotide sequence data are becoming available for analysis. There is an urgent need for development of theoretical tools to analyze the genome data, including identification of protein-coding sequences. The majority of the methods developed to date require prior information about the genome before accurate predictions can be made. Because such information is not available for many parasites, these methods cannot be directly applied. In this article, Alok Bhattacharya and colleagues describe some of the gene-prediction methods commonly in use, and a new method, GeneScan, that they have developed for the analysis of parasite genomes. PMID- 10689335 TI - No NO production during human Toxoplasma infection. PMID- 10689337 TI - Reply. PMID- 10689336 TI - Why do malaria parasites sequester? PMID- 10689338 TI - Toxoplasma gondii and Overdulve. PMID- 10689339 TI - Chloroquine binds in the cofactor binding site of Plasmodium falciparum lactate dehydrogenase--a response. PMID- 10689340 TI - Reply. PMID- 10689341 TI - Voices in my head. PMID- 10689342 TI - Seeing in three dimensions: the neurophysiology of stereopsis. AB - From the pair of 2-D images formed on the retinas, the brain is capable of synthesizing a rich 3-D representation of our visual surroundings. The horizontal separation of the two eyes gives rise to small positional differences, called binocular disparities, between corresponding features in the two retinal images. These disparities provide a powerful source of information about 3-D scene structure, and alone are sufficient for depth perception. How do visual cortical areas of the brain extract and process these small retinal disparities, and how is this information transformed into non-retinal coordinates useful for guiding action? Although neurons selective for binocular disparity have been found in several visual areas, the brain circuits that give rise to stereoscopic vision are not very well understood. I review recent electrophysiological studies that address four issues: the encoding of disparity at the first stages of binocular processing, the organization of disparity-selective neurons into topographic maps, the contributions of specific visual areas to different stereoscopic tasks, and the integration of binocular disparity and viewing-distance information to yield egocentric distance. Some of these studies combine traditional electrophysiology with psychophysical and computational approaches, and this convergence promises substantial future gains in our understanding of stereoscopic vision. PMID- 10689343 TI - Dynamical approaches to cognitive science. AB - Dynamical ideas are beginning to have a major impact on cognitive science, from foundational debates to daily practice. In this article, I review three contrasting examples of work in this area that address the lexical and grammatical structure of language, Piaget's classic 'A-not-B' error, and active categorical perception in an embodied, situated agent. From these three examples, I then attempt to articulate the major differences between dynamical approaches and more traditional symbolic and connectionist approaches. Although the three models reviewed here vary considerably in their details, they share a focus on the unfolding trajectory of a system's state and the internal and external forces that shape this trajectory, rather than the representational content of its constituent states or the underlying physical mechanisms that instantiate the dynamics. In some work, this dynamical viewpoint is augmented with a situated and embodied perspective on cognition, forming a promising unified theoretical framework for cognitive science broadly construed. PMID- 10689344 TI - Visually guided collision avoidance and collision achievement. AB - To survive on today's highways, a driver must have highly developed skills in visually guided collision avoidance. To play such games as cricket, tennis or baseball demands accurate, precise and reliable collision achievement. This review discusses evidence that some of these tasks are performed by predicting where an object will be at some sharply defined instant, several hundred milliseconds in the future, while other tasks are performed by utilizing the fact that some of our motor actions change what we see in ways that obey lawful relationships, and can therefore be learned. Several monocular and binocular visual correlates of the direction of an object's motion relative to the observer's head have been derived theoretically, along with visual correlates of the time to collision with an approaching object. Although laboratory psychophysics can identify putative neural mechanisms by showing which of the known correlates are processed by the human visual system independently of other visual information, it is only field research on, for example, driving, aviation and sport that can show which visual cues are actually used in these activities. This article reviews this research and describes a general psychophysically based rational approach to the design of such field studies. PMID- 10689345 TI - Retrieval processing and episodic memory. AB - The emergence of brain imaging has had a major impact on research into the cognitive and neural bases of human memory. An area in which this impact has been particularly strong is retrieval processing - the processes engaged when attempting to retrieve information during a memory test. Several different classes of retrieval process - such as 'mode', 'effort' and 'success' - have been invoked to account for findings from neuroimaging studies of episodic retrieval. In this article we discuss how these different kinds of process, along with a fourth kind associated with 'retrieval orientation', can be investigated in brain imaging experiments. We then review studies of retrieval processing, and assess how well their designs match up to our proposed criteria for dissociating the neural correlates of different classes of retrieval process. We conclude that few studies have used designs that permit these different kinds of process to be independently identified, and that presently there is little evidence to indicate which kinds of processing can be fractionated in terms of their neural correlates. PMID- 10689346 TI - The Turing Test: the first 50 years. AB - The Turing Test, originally proposed as a simple operational definition of intelligence, has now been with us for exactly half a century. It is safe to say that no other single article in computer science, and few other articles in science in general, have generated so much discussion. The present article chronicles the comments and controversy surrounding Turing's classic article from its publication to the present. The changing perception of the Turing Test over the last 50 years has paralleled the changing attitudes in the scientific community towards artificial intelligence: from the unbridled optimism of 1960s to the current realization of the immense difficulties that still lie ahead. I conclude with the prediction that the Turing Test will remain important, not only as a landmark in the history of the development of intelligent machines, but also with real relevance to future generations of people living in a world in which the cognitive capacities of machines will be vastly greater than they are now. PMID- 10689347 TI - Finding the genes that direct mammalian development : ENU mutagenesis in the mouse. AB - The genetic control of mammalian embryogenesis is not well understood. N-ethyl-N nitrosourea (ENU) mutagenesis screens in the mouse provide a route to identify more of the genes that are required for mammalian development. The characterization of ENU-induced mutations can build on the resources provided by the mouse and human genome projects to help define the tissue interactions and signaling pathways that direct early mammalian development. PMID- 10689348 TI - The cancer genome anatomy project: building an annotated gene index. PMID- 10689349 TI - Protein-length distributions for the three domains of life. PMID- 10689350 TI - Regulation of adjacent yeast genes. PMID- 10689351 TI - Complex evolution of the inositol-1-phosphate synthase gene among archaea and eubacteria. PMID- 10689352 TI - Canine genetics comes of age. AB - The dog, as human's favored companion, is unique among animal species in providing new insights into human genetic disease. In this review, we will discuss both the breed and the population structure of dogs and why that makes canines amenable to genetic studies. We will review the current state of the map and discuss the particular disease states in which canines stand to make the greatest contribution to medical genetics. PMID- 10689353 TI - Endoderm development: from patterning to organogenesis. AB - Although the ectoderm and mesoderm have been the focus of intensive work in the recent era of studies on the molecular control of vertebrate development, the endoderm has received less attention. Because signaling must occur between germ layers in order to achieve a properly organized body, our understanding of the coordinated development of all organs requires a more thorough consideration of the endoderm and its derivatives. This review focuses on present knowledge and perspectives concerning endoderm patterning and organogenesis. Some of the classical embryology of the endoderm is discussed and the progress and deficiencies in cellular and molecular studies are noted. PMID- 10689354 TI - Twins. Novel uses to study complex traits and genetic diseases. AB - The challenge faced by research into the genetic basis of complex disease is to identify genes of small relative effect against a background of substantial genetic and environmental variation. This has focused interest on a classical epidemiological design: the study of twins. Through their precise matching for age, the common family environment and background environmental variation, studying diseases in non-identical twins provides a means to enhance the power of conventional strategies to detect genetic influence through linkage and association. The unique matching of identical twins provides researchers with ways to isolate the function of individual genes involved in disease together with approaches to understanding how genes and the environment interact. PMID- 10689355 TI - Circadian rhythms: new functions for old clock genes. AB - The mechanisms of circadian clocks, which time daily events, are being investigated by characterizing 'clock genes' that affect daily rhythms. The core of the clock mechanism in Drosophila, Neurospora, mammals and cyanobacteria is described by a transcription-translation feedback-loop model. However, problems with this model could indicate that it is time to look at the functions of these genes in a different light. Our a priori assumptions about the nature of circadian clocks might have restricted our search for new mutants in ways that prevent us from finding important clock genes. PMID- 10689356 TI - Selective peptidomimetic blockers of autoantigen presentation: a novel therapeutic approach to autoimmune disease. PMID- 10689357 TI - Urotensin II: fish neuropeptide catches orphan receptor. PMID- 10689358 TI - G-protein-coupled receptors: what limits high-affinity agonist binding? PMID- 10689359 TI - Reply PMID- 10689360 TI - Nematode genome sequence dramatically extends the nuclear receptor superfamily. AB - Nuclear receptors represent a large class of ligand-activated transcriptional regulators; about 70 members of this protein family have been cloned from mammalian or insect species. Thus, it came as a great surprise when the recent completion of the Caenorhabditis elegans genome revealed at least 228 genes for nuclear receptors. Clearly, some of these receptors are homologues of known receptors, but most lack homologues in other species. Whether these receptors possess homologues in mammalian species is of great interest; if these do exist, the size of the nuclear receptor superfamily could also expand dramatically in humans. PMID- 10689361 TI - How well can molecular modelling predict the crystal structure: the case of the ligand-binding domain of glutamate receptors. AB - The concept that the ligand-binding domain of vertebrate glutamate receptor channels and bacterial periplasmic substrate-binding proteins (PBPs) share similar three-dimensional (3D) structures has gained increasing support in recent years. On the basis of a dual approach that included computer-assisted molecular modelling and functional studies of site-specific mutants, theoretical 3D models of this domain have been proposed. This article reviews to what extent these models could predict the crystal structure of the ligand-binding domain of an ionotropic glutamate receptor subunit recently determined at high resolution by X ray diffraction studies. PMID- 10689362 TI - Development of pharmacological agents for targeting neurotrophins and their receptors. AB - Neurotrophins comprise a family of protein growth factors that control the survival, growth, and/or differentiation of neurons and several other cell populations derived from the neuroectoderm. Neurotrophins and their receptors are important targets for the therapy of human disease, with potential applications ranging from the treatment of chronic or acute neurodegeneration to pain and cancer. Neurotrophins have been used clinically but are poor pharmacological agents. Consequently, approaches to develop pharmacological agents that target neurotrophins, their receptors or neurotrophin signaling pathways have been attempted. PMID- 10689363 TI - Cell-penetrating peptides. AB - The established view in cellular biology dictates that the cellular internalization of hydrophilic macromolecules can only be achieved through the classical endocytosis pathway. However, in the past five years several peptides have been demonstrated to translocate across the plasma membrane of eukaryotic cells by a seemingly energy-independent pathway. These peptides have been used successfully for the intracellular delivery of macromolecules with molecular weights several times greater than their own. Cellular delivery using these cell penetrating peptides offers several advantages over conventional techniques because it is efficient for a range of cell types, can be applied to cells en masse and has a potential therapeutic application. PMID- 10689364 TI - Protease-activated receptors: sentries for inflammation? AB - Cell-surface protease-activated receptors (PARs) appear to have evolved to detect extracellular enzymatically active serine proteases such as trypsin and thrombin. The predominant location of PARs on endothelia and epithelia and the discovery of enzymes such as trypsin within these tissues, together with the linkage of PARs to cytoprotective pathways, provide new information on autocrine and paracrine signalling within these critical barriers. In this article, the ways in which the distribution and function of PARs could be harnessed by pharmacologists as novel anti-inflammatory therapeutic strategies are discussed. PMID- 10689365 TI - Galanin receptor subtypes. AB - The neuropeptide galanin, which is widely expressed in brain and peripheral tissues, exerts a broad range of physiological effects. Pharmacological studies using peptide analogues have led to speculation about multiple galanin receptor subtypes. Since 1994, a total of three G-protein-coupled receptor (GPCR) subtypes for galanin have been cloned (GAL1, gal2 and gal3). Potent, selective antagonists are yet to be found for any of the cloned receptors. Major challenges in this field include linking the receptor clones with each of the known physiological actions of galanin and evaluating the evidence for additional galanin receptor subtypes. PMID- 10689366 TI - [ [In Process Citation] PMID- 10689367 TI - [Yellow fever]. PMID- 10689368 TI - [Species of Anopheles (Culicidae, Anophelinae) in a malaria-endemic area, Maranhao, Brazil]. AB - INTRODUCTION: The study of the seasonal fluctuation, nocturnal activity, relative abundance and the richness of Anopheles species in anthropic environment is essential to the understanding of the their bioecology and to the surveillance program of malaria control. METHODS: The Anopheles species were studied from 6 P.M. to 6 A.M., once a month, for one year, from October 1996 to September 1997, in the municipal district of Raposa, of the Sao Luis island, Maranhao state. The basic method was the capture of female specimens on human baits in peri and intradomicile sites by means of aspiration tube and guided luminous focus. RESULTS: A total of 1.407 specimens were collected and distributed as follow: Anopheles aquasalis (82% of the sample), Anopheles galvaoi (10,2%), Anopheles albitarsis (6,4%), Anopheles evansae, Anopheles nuneztovari, and Anopheles triannulatus davisi (the last three represented together 1, 4%). The anophelines occurred all year round, mainly in the rainy period, being more frequent in the intra (75,3%) than in the peridomicile site (24,7%), showing a clear preference to suck blood in the evening. CONCLUSION: The behavioural variation of Anopheles shows that the different species are becoming adapted to closeness to human habitations, in the rural zone of the Sao Luis island. PMID- 10689369 TI - [Taste preference for sweets and caries prevalence in preschool children]. AB - OBJECTIVE: To assess the preference for sweetness among preschool children and differences between less and more deprived groups. In addition, to assess whether sweet taste preference was associated with presence of caries. METHODS: The sample was composed by 572 preschool children aged between 4 and 6, distributed in three day nurseries of varied socioeconomic background. Cross-sectional study developed in two steps. Preference for sweetness was assessed using a modified version of the Sweet Preference Inventory. The solutions varied in sugar concentration from 0 to 1,17 molar (0 to 400 g / litre). The presence of caries was assessed using the defs index. The socioeconomic status of the sample was classified according to the origin of domicile. RESULTS: The variation in preference for sweetness in our sample was too small. Most children preferred the sweetest juice. This reduced the ability of this variable to explain variation in caries prevalence. Despite this limitation, our results showed that socioeconomic level influenced preference for sweetness, which in turn was associated with caries prevalence. CONCLUSION: The socioeconomic status influence the sweetness preference and this, in turn, is associated with the dental caries prevalence. PMID- 10689370 TI - [Evaluating a support program for children victim of domestic violence]. AB - INTRODUCTION: Domestic violence against children has been a subject of concern to many Brazilian institutions, though there are few studies about the services offered to the community. The aim of this study is to evaluate administrative and operational aspects of the SOS Crianca and provide insight to similar programs with comparative data. METHODS: A cross-sectional study of 976 data sheets of investigated cases until 1993 was conducted. Study variables were: characteristics of the support program required, proceedings to investigate reported cases, follow-up duration of reported cases, and referrals to other institutions. RESULTS: Out of 976 data sheets analyzed, 587 involved domestic violence against children: 38.7% cases of physical abuse, 27.7% of neglect, 26.3% of psychological abuse and 7.3% of sexual abuse. Most of the complaints (32.5%) came from family members of victimized children. The program's investigation process took between 126 to 212 days. Each investigated case demanded an average of 2.7 to 4.6 procedures. The majority of the cases (44.0%) were referred to a law court. This study emphasizes the need of a computerized database for optimizing the services provided and the victims' follow-up. It also suggests the need of ongoing staff training and development of broader and stronger connections to the social and health services. PMID- 10689371 TI - [American cutaneous leishmaniasis in the State of Acre, Brazil]. AB - OBJECTIVE: Present a statistical survey of American Cutaneous Leishmaniasis (ACL) in the State of Acre. METHODS: Data were obtained from the forms of the "Campaign against Leishmaniasis", from January 1992 to December 1997. Descriptive statistical analysis was applied. RESULTS: There were 2.557 registered cases. The highest prevalence was found at the microregion of Brasileia (231.8 cases/10,000 inhab.). The predominant clinical form was cutaneous (84.05%). The disease occurred mostly among males (71.02%). Half of the cases were among people with 24 years of age or younger. Most cases were people with rural occupations. 83.97% of the cases were diagnosed by clinical examination. The longest period to seek medical treatment was registered in the mesoregion of Jurua Valley (10.37 months) CONCLUSION: The high number of cases suggests that it might be necessary to study the psychosocial implications of the disease and identify factors contributing to the delay in treatment. PMID- 10689372 TI - [Mosquitoes potential vectors of canine heartworm in the Northeast Region from Brazil]. AB - INTRODUCTION: In some coastal districts of Sao Luis, capital of the state of Maranhao, Brazil, the prevalence of Dirofilaria immitis is more than 40% in house dogs. Natural potential vectors, as found in other areas of Northeastern Brazil, are unknown. The aim of this study was to identify probable vectors of the disease. METHODS: Mosquito catches were performed at a coastal, district Olho d'Agua, in S. Luis, to look for local potential vectors. Captures were carried out monthly, from March 1996 to May 1997, outdoors, having a man and a dog as baits. Mosquitoes were dissected for D. immitis larvae. RESULTS: A total of 1,738 mosquitoes belonging to 11 species were collected. Culex quinquefasciatus, the only species collected every month, was more frequently in the dry season. It accounted for 54.5% of the total, followed by Aedes albopictus (20. 3%), Ae. scapularis (11%) and Ae. taeniorhynchus (11%). D. immitis larvae were detected in 0.1% of the Cx. quinquefasciatus dissected (L3 in the Malpighian tubules) and 0.5% of the Ae. taeniorhynchus (L2 in the Malpighian tubules). CONCLUSION: Ae. taeniorhynchus and Cx. quinquefasciatus are considered natural potential vectors of the canine heartworm in Sao Luis. The role of Cx. quinquefasciatus as primary vector of D. immitis, however, needs further evaluation. PMID- 10689373 TI - [Dengue fever: a post-epidemic sero-epidemiological survey in an urban area setting at a northwestern county of Sao Paulo State, Brazil]. AB - OBJECTIVE: The objective of this study was to evaluate the real size of the epidemics registered in the urban area of the county of Santa Barbara D'Oeste, SP, Brazil, from April to June, 1995. The measurement of the epidemiological validity of the official surveillance system criteria and its positive predicted value were adopted as specific goals. METHODS: A sero-epidemiological survey was carried out over a sample of 1,113 sera from citizens of Santa Barbara D'Oeste, through a systematic random sampling of houses, five months after the end of the epidemics. Infection rates were compared with the infestation indexes by Aedes aegipty and the notified cases amongst the county sections. The importance of submitting patients with clinical suspicion of dengue to laboratory tests was discussed. RESULTS AND DISCUSSION: It was found that infection rates by dengue virus varied in the same direction and proportion as the presence of Aedes aegipty larvae reported by the "Breteau Index", as well as the number of cases reported by the official notifiable diseases surveillance system during the epidemics. A prevalence of 630 by 100 thousand inhabitants was found, a 15-fold rate when compared to the laboratory positive sera from cases detected by the surveillance system during the epidemics. A retrospective comparison with the surveillance reports, using serological results as a gold standard, also showed that the majority of dengue specific serum-positive individuals were not detected during the epidemics, otherwise cases that did not present serological reaction were notified exhibiting a low positive predictive value of clinical diagnosis (15,6). PMID- 10689374 TI - [Low prevalence of bodyweight-for-height deficit: comparison of stunned and no stunned Brazilian children]. AB - OBJECTIVES: The aim of this study was to investigate the relationship between abdominal circumference and weight-for-height in children. The average of 18 anthropometric and body proportionality indexes were compared among four groups of children: stunted and non-stunted Brazilians, Peruvians and North-Americans. METHODS: There were studied 386 children aged 6-59 months living in a poor neighborhood in Pelotas, Brazil. Anthropometric measurements (weight, recumbent length or height, sitting height or crown-rump length; head, chest, upper arm and abdominal circumferences; triceps, biceps, subescapular and suprailiac skinfold thickness; biacromial and biiliac breadths) were obtained. Muscle, fat, total upper arm areas, leg length and body proportionality indexes were calculated. RESULTS AND CONCLUSIONS: The study sample showed high levels of morbidity, low parental educational levels, poor access to health services and poor housing conditions. Stunted Brazilian children had lower means for most of the anthropometric measurements when compared to non-stunted Brazilians and North American children. However, stunted children showed larger abdominal, head and thoracic circumference in relation to their stature than non-stunted children. The low prevalence of weight-for-height among the children of this study is not a result of excess of fat or muscle tissue, and may be partly explained by an increase in head and trunk dimensions (including abdominal circumference) relative to the child's stature. PMID- 10689375 TI - [Referred morbidity and care seeking on mental disorders in childhood and adolescence]. AB - INTRODUCTION: Determining the prevalence of mental disturbances in childhood and adolescence gives us a better knowledge of their distribution in a given age group and provide us data for planning, implementing and evaluating health care programs. This survey was centered on complaints of "nervous problems" in a population group ranging from 1 to 19 years old as a tool to measure mental illness prevalence in that age group. METHODS: A group of 141 children and teenagers with complaints of nervous problems participated in the study, drawn by applying questionnaires from June, 1989 to July, 1990 to a sample of 3,158 people in the age group 1-19 years old, living in the Southeast area of Grande Sao Paulo. It was conducted an analysis of the nature of the complaints, their referred reasons and behavior, age, and gender. RESULTS: The prevalence of complaints of nervous problems was 4.7%. The older they were the more they complained. It was noticed a male predominance in the younger group and female preponderance in the group 14 years old or older. One in five tried to get any help, and severity of their complaint was the most important predictor for that. CONCLUSION: The prevalence was due to the population's ability to identify mental illnesses and it also reflects the family understandings of these problems. As only a few sought for health care, possible causes were identified and intervention actions were proposed to satisfy the unattended needs. PMID- 10689376 TI - [Mortality risk measure inequalities among workers in Southeast Brazil]. AB - INTRODUCTION: The main causes of illness and death in Brazil have been migrating backwards into the younger population during the last few years, increasing especially in the more productive age groups. Given the relationship between work and health/disease process, the hypothesis to be considered is that this phenomenon is partially due to the deterioration of workplace conditions. To contribute to investigating this hypothesis, this study estimates mortality risk indicators for the population of Botucatu, in the Southeast region of Brazil, classified according to their occupation. METHODS: Standardized mortality coefficient, standardized risk ratio, and years of potential life lost were calculated for the inhabitants of Botucatu who died after their 10th birthday, between January 1997 and March 1998, and classified according to their occupation and main cause of death. Occupational and medical information was obtained by interviewing families of the deceased and their doctors, and checking medical files. RESULTS: The standardized mortality coefficient ranged from 0.6 to 39.9 deaths/1000 workers in different occupations. The years of potential life lost ranged form 33 to 334 years/1000 workers. The ranking of causes of death varied according to occupation and the mortality risk considered. CONCLUSION: The risk measures analyzed showed a high heterogeneity when associated to occupation and causes of death, which reflects the great social inequality existing in the studied population. PMID- 10689377 TI - [Functional aging and work conditions in forensic workers]. AB - OBJECTIVE: The objective of this study was to evaluate aging associated to work conditions. METHOD: Eight hundred and seven forensic workers answered the questionnaire "Work Ability Index - WAI". The ergonomic conditions were analysed using a job analysis method - AET (Rohmert & Landau). RESULTS: The largest number of employees were mainly submitted to cognitive demands at work. The most reported diagnosed diseases were: musculoskeletal diseases (and lesions), neurological (including emotional disturbances), respiratory, digestive, skin and cardiovascular diseases. The analysis of the logistic regression models showed that: female workers, those with longer time on the job and job title of operational helper, increase the odds ratio to present low or moderate WAI. DISCUSSION: The results point out the need to improve the working conditions. It is suggested the implementation of Specialized Safety Engineering and Occupational Medicine Service, as it is demanded by the Federal Law 6,514 of 1977. PMID- 10689378 TI - [Using food frequency questionnaire in past dietary intake assessment]. AB - This paper aims to discuss which one is the best estimator of past diet: a retrospective report or a recent diet recall. The analysis included 13 articles published between 1984-1997 and selected from a MEDLINE search and from other reviews on this subject. The selection criterion was the use of a food frequency questionnaire (FFQ) in a validation study of retrospective report of dietary intake in remote past. Literature review shows that even taking into account misclassification, retrospective report of diet usually yields to a more reliable estimate of past diet pattern than current report. Past diet recall was strongly influenced by current intake and by diet patterns change. Analyzed investigations indicate that agreement between original and retrospective report was higher either for foods eaten rarely or frequently and lower for foods moderately consumed. This review allows considering the FFQ as a valuable instrument when studying the role of diet on the etiology of chronic diseases. PMID- 10689379 TI - [First occurrence of Biomphalaria straminea in the South Goiano, Brazil]. AB - The objective of this note is to report the occurrence of the Biomphalaria straminea in the county of Cachoeira Dourada in the south of Goias, and with that enhance the knowledge about its geographical distribution in the state. More than identify the species, the collection of thirty samples helped study and verify the presence of cercariae, especially because they were found in a setting which offers proper conditions to the development of a focal transmission site of schistosomiasis. PMID- 10689380 TI - [Availability of voluntary prenatal HIV screening in primary health centers]. AB - A summary of the main steps towards the implementation of anti-HIV testing for pregnant women in the city is presented. Starting from August 1996, voluntary HIV testing became available to pregnant women seen at primary health centers in Ribeirao Preto, SP (Brazil), as part of the Prenatal Care Program. By the end of 1998, 68.3% of the 17,589 women seeking prenatal care had been tested, resulting in a positivity rate of 0.76%. PMID- 10689381 TI - [Isolation of rabies virus in Molossus ater in (Chiroptera: Molossidae)in Sao Paulo State, Brazil]. AB - This is a report of rabies infection in insectivorous bat Molossus ater in the city districts of Aracatuba, Penapolis and Sao Jose do Rio Preto, in Sao Paulo state, Brazil. Fluorescent antibody test detected the virus in the brain and isolation was obtained by intracerebral inoculation of mice with nervous tissue and organs suspension. There was no contact with humans or other animals. PMID- 10689382 TI - [Serological survey for American cutaneous leishmaniasis in stray dogs in the S. Paulo State, Brazil]. AB - A serological survey was made in 973 stray dogs caught near green areas in S. Paulo county by the indirect immunofluorescence test. No positive serum was found, however autochthonous human cases of ACL that occurred in the county show the circulation of the parasite in the environment. This fact, associated with the existence of the vectors and the apparent absence of infected dogs in the areas studied, suggest that stray dog plays an insignificant role in the spread of the parasite. The cycle of the parasite in Sao Paulo county has been maintained by wild animals, hence the dog would be an accidental host just as humans. PMID- 10689383 TI - Incorporation of ubiquitin into the rat brain mitochondria is accompanied by increased proteolytic digestibility of MAO. AB - Incubation of rat brain mitochondria with ubiquitin followed by mitochondria sedimentation was accompanied by reduction of ubiquitin content in the supernatant only when ATP was included into the incubation mixture. Subsequent incubation of resedimented mitochondria revealed higher sensitivity to trypsin of MAO-A in ubiquitin-incorporated mitochondria. In control mitochondria (initially incubated with ATP) 0.5 mg/ml trypsin caused a decrease of MAO-A activity by 32.2 +/- 4.2%, whereas in ubiquitin-incorporated mitochondria (initially incubated with ATP + ubiquitin) reduction of MAO-A activity was significantly higher (51.4 +/- 2.5%, p < 0.02). Activity of MAO-B was resistant to trypsinolysis and incorporation of ubiquitin did not influence sensitivity of MAO-B to trypsin. Although there is no direct evidence yet that mitochondrial MAO is a target for ubiquitination the increased sensitivity to trypsinolysis of MAO-A suggests that incorporation of ubiquitin into mitochondria may increase susceptibility of MAO to certain proteases involved into degradation of these enzymes. PMID- 10689384 TI - Presence of SSAO in human and bovine meninges and microvessels. AB - In spite that SSAO enzyme is widely distributed in almost all tissues, specially in vascularized ones, its presence in brain microvessels is still controversy. Our results resolve this question showing that both human and bovine cerebrovascular tissues do contain the SSAO enzyme. This was achieved biochemically, using benzylamine and methylamine as substrates, and by immunoblot analysis, using polyclonal antibodies anti-SSAO that recognized a 100 kDa single band in tissue homogenates. PMID- 10689385 TI - Advances in evidence-based information resources for clinical practice. PMID- 10689386 TI - Tenets for physicians in the new millennium. PMID- 10689387 TI - Polymorphic tachycardia after cardiac arrest. PMID- 10689388 TI - A man with fever, rigors, and poor oral hygiene. AB - A 62-year-old man presented to the emergency department with a one-week history of subjective fever and rigors. He had had epigastric pain for three weeks, for which he was taking ranitidine, and in the past two to three months had experienced night sweats, a nonproductive cough, nausea, vomiting, and a 30-lb weight loss. He denied dsypnea, chest pain, hematochezia, melena, or any change in bowel habits. PMID- 10689389 TI - Molecules of the brain. AB - Progress against a range of brain disorders is being sustained by the use of genetic research techniques to identify specific molecules involved in brain disease, and by the realization that the identified molecules may disclose novel therapeutic targets. Both strategies are illustrated by recent insights and interventions in Alzheimer's disease and Parkinson's disease. PMID- 10689390 TI - Management of rapidly progressive glomerulonephritis. PMID- 10689391 TI - Cardiac syndrome X: an overview. AB - Not all patients with angina have myocardial ischemia. A sizable minority--up to 30% of angina patients studied at tertiary referral centers--have normal coronary angiograms. Such patients often undergo an expensive and extensive array of testing and treatment. Yet the prognosis is generally good, and symptomatic management may be effective. PMID- 10689392 TI - Chronic leg ulcers: types and treatment. AB - Disorders of the arteries, veins, or nerves, alone or in combination, can result in leg ulcers. The presentation in these cases varies with the cause, which in turn guides management. A differential diagnosis is critical, because treatment that is essential for one type of ulcer may be contraindicated in another. PMID- 10689393 TI - Improving management of type 2 diabetes mellitus: 6. Chromium. PMID- 10689394 TI - Coping with change: 4. Clear definitions and creative solutions. PMID- 10689395 TI - Nurses are not their sisters' keepers. PMID- 10689397 TI - The enigmatic nursing workforce. PMID- 10689396 TI - Climate, culture, context, or work environment? Organizational factors that influence nursing practice. PMID- 10689398 TI - Analysis of patient profile in predicting home care resource utilization and outcomes. AB - OBJECTIVES: The study identifies patient profile variables that explain variation in resource utilization and outcomes for home healthcare. BACKGROUND: The healthcare reform and the demand for quality patient care have increased the need to identify key patient characteristics that can predict the use of resources and outcomes; however, home healthcare industry currently lacks adequate information collection to reflect these needs. This study explored both the resource use and care outcomes for nursing administrators in monitoring quality, resource distribution, and reimbursement policy decision making. METHOD: The conceptual framework is based on Donabedian's quality care elements (structure, process, and outcome) and Nursing Minimum Data Set. This is a retrospective descriptive study design in which 244 patient records and data were obtained from a home healthcare agency located in Washington, DC. A series of stepwise and discriminant analyses was conducted for data analysis. RESULTS: The findings indicated that the total number of nursing diagnoses and two specific nursing diagnoses (alteration in mobility and knowledge deficit in i.v. therapy) were strong predictors of overall resource use. Prognosis proved to be the strongest predictor of discharge outcomes. CONCLUSIONS/IMPLICATIONS: The results indicated that data related to nursing diagnoses and nursing interventions can provide valuable information in predicting resource use. Prognosis made by nursing judgment was also sensitive in predicting patient outcomes. These critical data elements should be included in describing home health patient characteristics and related resource utilization and care outcomes. PMID- 10689399 TI - The impact of systems redesign on staff, patient, and financial outcomes. AB - OBJECTIVES: The purpose of this study was to measure the impact of a change initiative using the strategies of inpatient bed consolidation and patient population reaggregation on staff, patient, and financial outcomes. BACKGROUND: Bed consolidation and patient population reaggregation are extensively used strategies in the hospital industry. However, the state of science as it relates to these two strategies is limited, with no studies measuring the effect of bed consolidation, patient reaggregation, or both in isolation of multiple other concurrent care model changes. METHODS: An exploratory, single case, longitudinal field study design with embedded levels of analysis was used for the study. The impact associated with the reaggregation and consolidation of the acute inpatient system of care was evaluated at the organization, the acute inpatient nursing unit, and the individual nursing staff member levels. This article reports the outcomes found at both the nursing unit level and the staff member level on three general medicine nursing units. Baseline nursing unit-level data are reported from fiscal year 1994-1995 and the first half of fiscal year 1995-1996 (July 1995 through December 1995); data from January 1996 through June 1996 reflect the planning phase of the project. Outcomes related to the change process are reported from the time period January 1996 through June 1997. Staff perceptions of multiple variables were measured at five different data points over a 2-year period. Both quantitative and qualitative data were collected. RESULTS: Unit level cost and hours per patient day (HPPD) increased during or immediately after the merger of major patient populations on two of the study units; no significant variation was found in medication errors or patient falls. The quantitative and qualitative data analysis of the nursing staff member surveys revealed a nursing staff that was dissatisfied with many aspects of their job, worried about job security, had low morale, and had many concerns about the quality of care provided to patients. CONCLUSIONS: This study suggests that consolidation and reaggregation strategies may cause an increase in costs and HPPDs, at least on a short-term basis. Also, these strategies may have a significant effect on the morale and job satisfaction of unit-level nursing. In light of these findings, on going testing of the efficacy of consolidation and patient reaggregation strategies in improving quality and cost outcomes is essential. PMID- 10689400 TI - Patient-centered documentation: an effective and efficient use of clinical information systems. AB - After 2 years of experience with a basic computerized system for documenting care, the nursing leaders in an inner city hospital undertook a redesign process to create an effective and efficient system for documentation that also provided data for monitoring care processes, patient outcomes, and staff performance. The basic system was a source of frustration and dissatisfaction for nurses, physicians, and managers. The redesigned system has exceeded the expectations of staff and physicians and delighted managers. Managers can now access the clinical data collected by the staff to create meaningful reports describing the patient population, identifying patients needs, monitoring staff compliance with care standards, and tracking improvements in care processes. PMID- 10689401 TI - Assessment: the first step in creating a more integrated system-wide approach to nursing practice. AB - As complex healthcare systems are created, professional nurses in these systems must address the integration of nursing practice. To facilitate the transition to a more common system-wide approach to professional practice, leaders at one health system began by conducting an assessment of their current reality of nursing practice. The authors describe the assessment process, including the staff survey questionnaire and recommendations to strengthen nursing practice and make the transition to a more common system-wide approach to professional practice. PMID- 10689402 TI - Reducing the use of physical restraints in nursing homes. Regulatory harassment or good medicine? PMID- 10689403 TI - Validity of malingering signs questioned. PMID- 10689404 TI - Another cause of anterior knee pain. PMID- 10689405 TI - Need more power in your communication technology? Increasing bandwidth will improve speed and expand options. PMID- 10689406 TI - Generalized pruritus without primary lesions. Differential diagnosis and approach to treatment. AB - A 65-year-old man presented with recurrent generalized pruritus and excoriations of many years' duration. He had been treated with antihistamines, topical corticosteroids, and antibiotics for secondary wound infections, but improvement was only temporary. He had also been hospitalized for leg ulcers complicated by cellulitis. Examination revealed multiple oval and linear red papules and nodules measuring 0.5 to 2 cm in diameter. Some of the lesions were eroded and had a central crater and yellowish crust. The patient also had hypopigmented linear scars localized to the posterior scalp, neck, upper back, chest, abdomen, arms, and legs with sparing of the middle and lower back (figures 1 and 2). An ulcer measuring 1.5 x 2 cm that was surrounded by indurated skin was present on the medial aspect of his right ankle. The ulcer was partially covered by yellow exudate. There was no evidence of cellulitis. Liver enzyme, serum creatinine, and thyrotropin levels, as well as a chest roentgenogram, were normal. Wound cultures for bacteria and fungi were nonsignificant. A punch biopsy from a representative lesion showed an abrupt epidermal defect with sparse superficial lymphocytic infiltrate in the dermis. The patient was admitted to the hospital to isolate him from his home environment. He received a 10-day course of systemic cephalexin, topical clobetasol propionate ointment for the affected skin areas, and oral hydroxyzine for pruritus. Ultraviolet light therapy was instituted once daily and was to continue for 2 months. His lesions had improved moderately by the time he was discharged from the hospital. On follow-up 2 weeks later, his lesions were flat and had resulted in hypopigmented scars. Three months later, however, he had persistent, intense pruritus, and new excoriations had developed on his forearms and back. He improved after receiving treatment with oral doxepin hydrochloride. PMID- 10689407 TI - In-office diagnosis of exercise-induced asthma. PMID- 10689408 TI - Hypertension in patients with diabetes. Why is aggressive treatment essential? AB - Hypertension and diabetes are interrelated diseases. Alone, each condition is a risk factor for cardiovascular disease and, together, they strongly predispose to end-stage renal disease, coronary artery disease, and peripheral vascular and cerebrovascular disease. Pharmacologic treatment of hypertension can substantially reduce morbidity and mortality in diabetic patients with hypertension, but adequate control of blood pressure is seldom achieved in a clinical setting. More aggressive treatment is needed to improve the prognosis for this over-expanding patient population. PMID- 10689409 TI - Viral pneumonias. Infection in the immunocompromised host. AB - Three herpesviruses--herpes simplex, varicella-zoster, and cytomegalovirus- commonly cause respiratory tract infections in immunocompromised patients. Adenoviruses and measles virus are also significant causes of respiratory disease in this population. Diagnosis of herpesvirus infections is difficult because these viruses can establish latency and are often shed intermittently in the absence of invasive disease. A positive respiratory tract culture of herpesviruses alone is not diagnostic of active invasive disease. Preventive measures should focus on limiting the patient's exposure to active infection, broad use of available vaccines in children and susceptible adults, and use of hyperimmune globulin and chemoprophylaxis in high-risk patients. Adenovirus pneumonia is diagnosed by viral culture and rapid antigen detection assays, whereas measles pneumonia is often identifiable by the characteristic rash. Treatment of either adenovirus or measles pneumonia is primarily supportive. PMID- 10689410 TI - Non-Q wave myocardial infarction. Assessment and management of a unique and diverse subset. AB - Acute cardiac events involving coronary symptoms, elevated enzyme levels, and electrocardiographic changes without the development of Q waves often result in higher rates of reinfarction and unstable angina than do more severe myocardial infarctions. The incidence of these non-Q wave events is on the rise, possibly because of earlier detection and treatment of heart disease. Familiarity with the characteristics and management of the condition, therefore, is more important than ever. PMID- 10689411 TI - Abnormal findings on liver function tests. Interpreting results to narrow the diagnosis and establish a prognosis. AB - Evaluating abnormal liver test results requires careful attention to the corresponding clinical data obtained during history taking and physical examination. Generally, it is helpful to separate liver tests into three categories: tests that assess synthetic function, tests that assess hepatocellular necrosis (hepatocellular enzymes), and tests that assess cholestasis. The clinical setting together with the specific pattern of liver function abnormalities can narrow differential diagnosis and provide a cost effective approach to assessing patients and identifying those who need liver biopsy. PMID- 10689412 TI - Tests for acute and chronic viral hepatitis. Finding your way through the alphabet soup of infection and superinfection. AB - Because clinical signs are of little or no help for identifying various causes of viral hepatitis, accurate diagnosis can only be achieved with serologic and molecular testing. Knowledge of the strengths and limitations of these tests allows rational use and interpretation of results. The findings have implications for public health surveillance, estimating prognosis, and identifying candidates for treatment. PMID- 10689413 TI - Antiviral therapy for chronic hepatitis B and C. Which patients are likely to benefit from which agents? AB - As primary care physicians become increasingly involved in diagnosis and treatment of patients with chronic viral hepatitis, an understanding of the antiviral options available, their limitations, and their side effects takes on a special importance. For chronic HBV infection, interferon alfa-2b requires only a 4-month course. However, it has adverse effects and contraindications and does not produce a universal response. Another option for HBV infection is lamivudine, which is administered orally and causes few side effects. However, relapse may occur when treatment is discontinued, and mutant virus may emerge. For chronic HCV infection, interferon alfa-2a, interferon alfa-2b, consensus interferon, and interferon combined with ribavirin have been used. The combination alternative is emerging as the method of choice in patients who do not have contraindications to oral ribavirin. Adverse effects are common, and durability of response varies according to HCV RNA level and genotype. PMID- 10689414 TI - Genetic liver disease in adults. Early recognition of the three most common causes. AB - The most common clinically important genetic diseases leading to liver dysfunction in adults are Wilson's disease, HHC, and alpha 1AT deficiency. Advances in molecular biology have led to the identification and characterization of the genetic defects in these conditions. Consequently, genetic testing for disease-causing mutations is now available for most of these disorders. However, it is important to understand the strengths and limitations of such testing. Genetic testing is probably most helpful in HHC because of the high frequency of the homozygous C282Y mutation among patients of northern European descent and the relatively high penetrance of the mutation with regard to clinical expression. Genetic testing is much less helpful in the other genetic liver diseases because of the high number of possible mutations and variable clinical expression. However, noninvasive phenotype-based screening tests and specific treatments are available for most genetic liver diseases. Appropriate use of screening tests in routine clinical practice can assist in early identification of genetic liver diseases and prevent development of end-organ damage. PMID- 10689415 TI - Side effects of antipsychotic drugs. Avoiding and minimizing their impact in elderly patients. AB - Antipsychotic drugs are very useful in treatment of psychosis and severe agitation in the elderly. Their use for other behavioral problems is contraindicated. Antipsychotics have many potential side effects (e.g., sedation, cardiovascular effects, anticholinergic effects, incontinence, reduced appetite, such motor disturbances as drug-induced parkinsonism, akathisia, dystonia, TD). Prevention, by using the minimum dose and duration of treatment possible, is the key to managing motor side effects. If prevention fails, drug-induced parkinsonism and dystonia may improve with use of anticholinergics, and akathisia may improve with use of benzodiazepines or low-dose propranolol. There is no proven treatment for TD, which is most likely to be observed during dose reduction or after discontinuation of antipsychotic drugs. Compared with older agents, newer antipsychotic drugs are less likely to cause parkinsonism, akathisia, and dystonia and may cause TD less often. More research is needed to clarify use of the new drugs in the elderly. PMID- 10689416 TI - Sexually transmitted diseases in women. Gonorrhea and syphilis. AB - Gonorrhea has been declining since its 1975 peak. Risk factors include age 15 to 19 years, multiple or casual sexual contacts, sexual activity related to drug use, and low socioeconomic status. Infection is usually mild but may be asymptomatic. While no physical signs are specific to the gonococcus, pelvic inflammatory disease is a common complication and cause of infertility and should be treated if it is suspected. Diagnosis of gonorrhea is typically by culture. Newer, more accurate tests are available but are more expensive. For treatment, the CDC recommends only highly effective regimens. Patients need to refer recent sexual partners for treatment and abstain from sexual intercourse until completion of therapy and resolution of symptoms. The incidence of syphilis appears to be declining in the United States, but it should be considered if an ulcer is found in the genital region. If untreated, the disease progresses through primary, secondary, latent, and tertiary phases, and systemic symptoms can mimic other conditions. Positive standard screening tests should be confirmed by fluorescent treponemal antibody absorption testing. Darkfield microscopy is appropriate for diagnosis of an ulcer. The treatment of choice for all phases of syphilis is a single dose of intramuscular benzathine penicillin. Other components of therapy include partner notification and patient follow-up. The spread of HIV is closely linked to STD transmission. Therefore, testing for HIV is strongly encouraged when another STD has been diagnosed. PMID- 10689417 TI - Evaluation of nursing home patients. A systematic approach can improve care. AB - As the US population ages, the proportion of patients receiving long-term care is increasing. To meet the challenge of providing quality care for these patients, physicians need to be prepared to efficiently evaluate their needs and formulate individualized care plans. In this article, Drs King and Lipsky discuss the unique aspects of caring for nursing home patients, including the role of patients' families in the overall plan. They present a practical, structured approach to evaluation and follow-up care, which they have encapsulated into two handy assessment forms. PMID- 10689418 TI - The physician's role in directing long-term care. Understanding the rules is important for protecting your patients and your practice. AB - Meeting the healthcare needs of patients who require long-term care presents many challenges, not the least of which are local and federal regulations, a formidable bureaucracy, and confusing paperwork. Despite these obstacles, long term care patients often are among those with the greatest need for well-trained physicians and high-quality medical services. This article reviews elements of long-term care, with an emphasis on how primary care physicians should order and provide services. PMID- 10689419 TI - A perplexing case of pruritic plaques. Bullous pemphigoid. PMID- 10689420 TI - Shattering the myths about male infertility. Treatment of male factors may be more successful and cost-effective than you think. AB - Male factors play a role in up to half of subfertile couples, contrary to the myth that male factors rarely play a role. In this article, Dr Sandlow counters this and other myths about male infertility and suggests that primary care physicians can increase a couple's chance of conceiving by evaluating for male as well as female factors. This article will also help primary care physicians provide appropriate education and treatment, as well as determine when to make a referral to a male-infertility specialist. PMID- 10689421 TI - Community-acquired pneumonia. Outpatient treatment of patients 16 years and older. Institute for Clinical Systems Improvement. PMID- 10689423 TI - The ABCs of hepatitis. PMID- 10689422 TI - Fever without source in children. Recommendations for outpatient care in those up to 3. AB - It is the author's goal to reduce risk to a minimum in children with fever without source at a reasonable cost with guidelines that are practical for office based physicians. Recommendations are as follows: All febrile infants and children up to 36 months of age who have toxic manifestations are to be hospitalized for parenteral antibiotic therapy after an expeditious evaluation of their condition that includes cultures of blood, urine, and cerebrospinal fluid. All febrile infants 7 days of age or less should be hospitalized for empirical antibiotic therapy after a complete evaluation for sepsis and meningitis has been done. Some low-risk febrile infants 8 to 28 days of age who appear well may be observed closely, either in hospital (with or without empirical antibiotic therapy) or as outpatients if the physician believes that close follow-up is ensured. Febrile infants 28 to 90 days of age should have an evaluation to determine whether they are in a low-risk group. Those not meeting low-risk criteria should be hospitalized for a complete "sepsis workup" and close observation, with or without empirical antibiotic therapy. Those who are considered low-risk can be treated as outpatients, as described, if close follow up is ensured. No laboratory tests or antibiotics are needed in a child over 90 days of age who has a temperature of less than 39 degrees C (102.2 degrees F) without identifiable source. A return visit is recommended if the child's fever persists for more than 2 to 3 days or if the condition deteriorates. A child with a fever of 39 degrees C or above can also be treated as an outpatient without antibiotics if close follow-up is ensured. Otherwise, a WBC count or ANC should be done. In those whose WBC count is 15,000/mm3 or more or whose ANC is 10,000 cells/mm3 or more, a blood culture should be done, and pending results, a single injection of ceftriaxone, 50 mg/kg, should be given. PMID- 10689424 TI - [Premolecular period of the postradiation remodelling of cells]. AB - Recovery of yeast cells after exposure to ionizing radiation was found in 1957. During the first decade, i.e. in the "premolecular period" of studying the phenomenon, its basic features were revealed: dependence on ploidy of cells, on their energy exchange, on radiation LTE, and others. A mathematical model of recovery was proposed; the damages causing death of irradiated haploid and diploid cells were shown to be double strand breaks of DNA. The concepts of universal biological importance of the cell property to repair genetic damages were formulated. PMID- 10689425 TI - [Dynamics of tritium content in flood-lands reservoirs of the Pripyat river and cooling pond of the Chernobyl nuclear plant]. AB - Tritium content in water from natural and artificial reservoirs within 30-km exclusion zone of the Chernobyl NPP has been determined. The increase of Tritium activity in the involved water reserwous has been registered in May 1994 and April 1995. As supposed the source of the increase, nuclear power plants, equipped with WWER reactors and located in catchment area of Pripyat river. PMID- 10689426 TI - [Assessment of lipid peroxidation and plasma membrane permeability for Ca2+ in the red blood cells of cattle after the long term grazing on radioactively contaminated territory]. AB - Ten years after the Chernobyl accident a physiological condition of cows was examined on radioactivy contaminated territory of the Novozibkov district of the Bryansk region. The long grazing of cattle on radioactivly contaminated territory revealed the increase in permeability of plasmatic membrane of the red blood cells to Ca2+ and the activation of process of lipid peroxidation. The sensitivity of the red blood cells of cows to incubations in hypertonic conditions was demonstrated. PMID- 10689427 TI - [Biological effects in natural populations of small rodents in radiation contaminated areas. The frequency of bone marrow polyploid cells of bank voles in different years following Chernobyl accident]. AB - On the basis of the metaphase analysis results, the peculiarities of dynamics of the genome mutation frequency (polyploid cells) were studied in bone marrow of bank voles inhabiting the areas with different radiocontamination level due to the Chernobyl accident (8-1526 kBq/m2 for 137Cs) in 1986-1991. Unexpectedly high frequencies of polypoid cells exceeding the pre-accidental level by a factor of 10(1)-10(3) were recorded in all populations studied. Relationship between the frequency of the parameter studied and the concentration of radionuclides incorporated in animal carcasses was proved. A statistically significant rise in the frequency of genome mutations with the time was revealed up to 1991, i.e. approximately to 12th post-accidental animal generation. PMID- 10689428 TI - [Features of action of low doses of gamma-radiation on yeast cells]. AB - Lethal effect of low doses and adaptive response to low doses of prolonged irradiation were investigated in experiments on yeast cells. The phenomenon of hypersensitivity at low dozes was not found in yeast cells at gamma-irradiation. The adaptive response was observed after exposure to low doses of prolonged irradiation, the degree of the reaction depends on a dose rate. The adaptive reaction was kept for some time after the termination of adaptive irradiation then the sensitivity of cells increased even in comparison with unirradiated ones. PMID- 10689429 TI - [Alteration of polymorphic systems of Centaurea scabiosa L. under chronic irradiation]. AB - Isoenzyme and morphological polymorphism alteration in populations of perennial grass Centaurea scabiosa L. (scaly cornflower) has been studied. These populations exist on the territory of East Ural Radioactive Trace more than 40 years and are chronically exposed to beta-irradiation. Directional shift of allele frequencies on the loci Per1, Pgi2, Sod1, Lap has been detected. The fact of accumulating genetic load by chronically irradiated populations has been demonstrated. Possible reasons of discovered alterations are discussed. The analysis of the obtained data shows that the irradiated populations have greater similarity with one another than with a control, but relation between genetic distances and accumulated doses hasn't been revealed. The hypothesis is that an extra factor--gene flow from a "clean" territory influences the genetic structure of irradiated populations. PMID- 10689430 TI - [DNA damage in cells exposed to ionizing radiation]. AB - Ionizing radiation induces variety of structural lesions in DNA of irradiated organisms. Their formation depends largely on the degree of cell oxygenation, the level of endogenous antioxidants, on DNA-protein complexes and compactization of DNA in the chromatin and activity of DNA repair systems. All ionizing radiation induced DNA lesions can arbitrarily be divided into two groups. Group 1 includes singly damaged sites (single-sites): base modification, single-strand breaks, alkaline-labile sites (including a basic sites). Group 2 contains: locally multiply damaged sites (clustered lesions), double-strand breaks, intermolecular cross-links. The yields of lesions of group 2 increases with high linear energy transfer of radiation and these lesions play a dominant role in the radiation death, formation of chromosome and gene mutations, cell transformation. PMID- 10689431 TI - [Concentration of T3 and T4 in blood of non-irradiated and irradiated with different doses rats fasted for two days beforehand]. AB - There were no changes in concentration of T3 and T4 in blood of the rats that were irradiated with a dose of 0.5, 1, 2, 4 and 6 Gy in comparison with non irradiated rats, if the animals were not fed for two days before decapitation. This suggests that the effect of ionizing radiation on thyroid function is mediated by anorexia syndrome. The decrease in concentration of T4 after exposure to 8 Gy cannot be explained by postradiative anorexia and most likely is connected with starting enterotoxemy in difficult cases of acute radiation sickness. PMID- 10689433 TI - [Adaptation to irradiation in vivo]. AB - The data about the increasing of radioresistance of cells and organism after the acute, chronic and prolonged irradiation in vivo were presented. The possible mechanisms of adaptation to irradiation connected with selection, stimulation of proliferative activity, forming of protecting system (protected proteins, antioxidant enzymes e.a.), activation of DNA repair and cAMP-, and Ca(2+) dependent phosphorylation systems were observed. The conclusion about complex mechanism of radiation adaptation was drawn. PMID- 10689432 TI - [Effect of ionizing radiation on the structure and functional properties of the basolateral membrane of small intestine enterocytes]. AB - The structural state and transport properties of basolateral membrane of rat small intestine enterocytes after exposure to X-ray irradiation (0.5; 1.0 and 2.0 Gy) were studied. The substantional suppression of the active Ca(2+)-transport process concomitant to versatile changes of the membrane structure involving the surface sites and intramembrane protein-lipid complexes was revealed one day after irradiation. Taking into account the early obtained data on apical membrane functional disorders these results confirm that ionizing radiation in sublethal doses induces the structure-function modification of enterocyte plasma membrane affecting the function of the small intestine epithelial cells. PMID- 10689434 TI - [The geterogenity of 137Cs and 90Sr distribution and dose loading on critical tissues of main seedling root]. AB - It is shown, that the roots of plants concentrate 137Cs and 90Sr from water solutions in different zones: 137Cs--mainly in a meristem zone, 90Sr--in a stretching zone. The similar character of radionuclide distribution was established regarding water and soil cultures. The real dose loading on critical tissues of main root have appeared to be much higher than it was expected from the assumption of uniform distribution of radionuclides in tissues. PMID- 10689435 TI - [Mechanisms and models of 137Cs migration in soils]. AB - The most spread mechanisms and models of 137Cs migration in soil were considered and the advantage of the models, which take into account the kinetics of sorption desorption above the models with two components were presented. PMID- 10689436 TI - [The analysis of coefficient of 137Cs transfer from soil into potato in relationship with 137Cs content in soil and plant mineral feed]. AB - The soil-potato transfer factor for 137Cs (TF) was estimated by using results of 137Cs activity concentration measurements in 214 samples of soil and potato taken at fields with various level of contamination with 137Cs. The relationships between the coefficient TF and soil characteristics (acidity pH (KCl), content of K2O, P2O5, CaO and MgO in soil) and soil contamination with 137Cs have been analysed. The results show that the TF values tend to decrease with increasing concentration of 137Cs, K2O, P2O5, and CaO in considered sod-podsolic sandyloam soil. The regression function describing the TF dependence of 137Cs, K2O, P2O5, and CaO content in soil has been derived. PMID- 10689437 TI - [Plant reaction to elevated ultraviolet irradiation: potential impacts of stratospheric ozone depletion]. AB - The ozone layer depletion evokes the increase of solar UV-B radiation intensity and corresponding reductions of growth (height, leaf area, fresh and dry weight), photosynthetic activity and flowering in higher plants. Competitive interactions also may be altered indirectly by differential growth responses. The UV-B sensitivity of plants varies both among species and among cultivars of a given species. Photosynthetic activity may be reduced by direct effects on the photosynthetic process or metabolic pathways, or indirectly through effects on photosynthetic pigments or stomatal function. Plants may also respond by accumulating UV-absorbing compounds in their outer tissue layers, which presumably protect sensitive target from UV-damage. The key enzymes in the biosynthetic pathways of these compounds are specifically induced by UV-B irradiation via gene activation. The effects of UV-B radiation on plants can be modified by prevailing microclimatic conditions. Plants tend to be less sensitive to UV-B under drought or mineral deficiency, while sensitivity increases under low levels of visible light. Prognoses of agricultural yield reduction and economic loss for different scenarious of stratospheric ozone depletion are presented. PMID- 10689438 TI - [UV-induced changes of structural and functional properties of blood lactate dehydrogenase isoenzymes in free state and in the presence of serotonin]. AB - Photoinduced changes of structural and functional properties of lactatedehydrogenase isoenzymes from human erythrocytes in free state and in the presence of serotonin have been studied by means of gel chromatography, electrophoresis, IR-spectrophotometry and by the method of definition of catalytic activity. UV-light influence induces photoinactivation of erythrocyte's LDH, while its inhibitory effect intensifies with the increase of irradiation dose. The complicated character of changes in electrophoretic mobility and percentage content of isoenzymes LDH-1, LDH-2, LDH-3 under the influence of UV rays testifies that the decrease of total enzyme activity of these isoforms in connected with their different photosensitiveness and represents the result of many-staged process which is characterized both by the consistent and parallel proceeding of its individual photochemical reactions. A pronounced photoprotective effect of serotonin towards the molecules of erythrocytic LDH isoenzymes has been discovered. It seems to be caused by formation of enzyme- biogenous amine complex affecting the secondary protein structure. PMID- 10689439 TI - [Biological effects of the low-intensive laser of the near infra-red spectrum]. AB - The immediate and the remote effects of the total one-time uniform irradiation with arsenid-hallium laser (wavelength 0.89 mu, pulse power 450 W, pulsed mode 80 3000 Hz) on the new-born Wistar rats have been examined. The complex of tests illustrated the biotropical effects of used laser regimes. The stimulating or braking of the animal's growth was observed in accodanse with the laser regime. PMID- 10689440 TI - [The international conference on human protection from electromagnetic fields hazard]. PMID- 10689441 TI - Mapping the human nervous system in health and disease. PMID- 10689442 TI - Kugelberg Lecture. Principles and pitfalls of nerve conduction studies. PMID- 10689443 TI - Lecture in honour of Professor Emeritus Fritz Buchthal. Acute and chronic demyelinating polyneuropathy: an overview. PMID- 10689444 TI - Utility of somatosensory evoked potentials (SEPs) in spinal cord lesions and functional surgery of pain and spasticity. PMID- 10689445 TI - Multimodal approaches in the evaluation of epilepsy patients for surgery. PMID- 10689446 TI - Movement-related brain electrical activity. PMID- 10689447 TI - New trends in magnetoencephalography. PMID- 10689448 TI - Cognitive potentials and corticocortical connections in humans. PMID- 10689449 TI - Motor cortex plasticity. PMID- 10689450 TI - What makes a brain oscillation abnormal? PMID- 10689451 TI - Electrophysiological and morphological changes in the peripheral nervous system with ageing. PMID- 10689452 TI - Clinical applications of cyclic alternating pattern. PMID- 10689453 TI - Power spectral sleep EEG findings in patients with obstructive sleep apnea and upper airway resistance syndromes. PMID- 10689454 TI - Survey of inherited peripheral nerve diseases. PMID- 10689455 TI - Interconnections between cortical areas revealed by transcranial magnetic stimulation. AB - The fact that TMS of cerebral cortex is associated with inhibitory as well as excitatory properties is important because it makes it possible to investigate interconnections between cortical areas and tracing these functional interconnections by a noninvasive excitation or inhibition and temporary interference with the flow of impulses in the cerebral cortex. An important tool is thereby added to the analysis of higher cortical functions. PMID- 10689456 TI - Isolation of late event-related components to checkerboard stimulation. PMID- 10689457 TI - Continuous EEG monitoring in the intensive care unit. PMID- 10689458 TI - Percutaneous stimulation of mechanoreceptors and peripheral neural transmission in normal subjects and patients with hereditary ataxias. PMID- 10689459 TI - Non-invasive pre-surgical evaluation with EEG/MEG source analysis. PMID- 10689460 TI - Excitability of the motor cortex in amyotrophic lateral sclerosis. PMID- 10689461 TI - Introducing priors in the EEG/MEG inverse problem. PMID- 10689462 TI - Spinal mechanisms of spasticity. PMID- 10689463 TI - Genotype-phenotype correlation in spinocerebellar ataxias (SCA). PMID- 10689464 TI - Cortical visual processing. PMID- 10689465 TI - Neuromagnetic recordings and magnetic brain stimulation in the evaluation of sensorimotor hand area interhemispheric differences: normative, experimental and patients' data. PMID- 10689466 TI - Does the vestibulo-ocular reflex use the same pathways for functions in roll and pitch planes? PMID- 10689467 TI - Short and middle latency vestibular evoked potentials to angular and linear acceleration. PMID- 10689468 TI - Evoked potentials in sound localization: timing of activity along the auditory pathway. PMID- 10689469 TI - EEG band centroid modifications in HIV patients. PMID- 10689470 TI - Genetic and antibody-mediated channelopathies at the neuromuscular junction. PMID- 10689471 TI - Mechanisms involved in the propagation of interictal epileptiform discharges in partial epilepsy. PMID- 10689472 TI - P300 and Alzheimer's disease: oddball task difficulty and modality effects. PMID- 10689473 TI - Value and limits of cardiovascular autonomic function tests. PMID- 10689474 TI - Non-invasive assessment of motor unit properties with linear electrode arrays. PMID- 10689475 TI - Electroencephalogram in metabolic encephalopathies. PMID- 10689476 TI - Evoked potentials to painful laser stimulation. PMID- 10689477 TI - Value and limits of expert systems in clinical neurophysiology: an integrated 'Avicennian' approach. PMID- 10689478 TI - Congenital muscular dystrophies. A short review of the recent progresses. PMID- 10689479 TI - Model-based delta plots beat power-based ones. PMID- 10689480 TI - Can evoked potentials be useful in monitoring multiple sclerosis evolution? PMID- 10689481 TI - Spinal reflex pathways transcend the scheme of an auxiliary or independent system. PMID- 10689482 TI - Characteristics of SEP from subthalamic nucleus in human. PMID- 10689483 TI - Spinal reflex studies enable to analyze supraspinal dysfunctions. PMID- 10689484 TI - Neurophysiological aspects of diagnosis in neuromuscular transmission defects--an update. PMID- 10689485 TI - Movement-related potentials and magnetic fields: new evidence for SMA activation leading MI activation prior to voluntary movement. PMID- 10689486 TI - Auditory information processing in comatose patients: EPs to synthesised 'musical' tones. PMID- 10689487 TI - High-rate transcranial magnetic stimulation: influence on short-term-memory, heart rate and blood pressure changes. PMID- 10689488 TI - Muscle sonography. PMID- 10689490 TI - MEG spontaneous activity in patients with memory disorders. PMID- 10689489 TI - Diagnostic relevance of anti-neural antibodies in dysimmune neuropathies. PMID- 10689491 TI - Inhibitory function in chronic focal epileptogenesis. PMID- 10689492 TI - Do GABAergic circuitries play a critical role in the regulation of seizure induced neuronal damage and synaptic reorganization in the rat hippocampus? PMID- 10689493 TI - Antiepileptic drugs potentiating GABA. PMID- 10689494 TI - Cognitive effects of GABAergic antiepileptic drugs. PMID- 10689495 TI - Cognitive impairment in multiple sclerosis: a longitudinal study. PMID- 10689496 TI - Event-related potentials in the assessment of cognitive function in multiple sclerosis. PMID- 10689497 TI - Electrophysiological investigations in multiple sclerosis dementia. PMID- 10689498 TI - Mechanisms of action of intravenous immunoglobulin (IVIg) in autoimmune and neuromuscular diseases. PMID- 10689499 TI - Chronic inflammatory demyelinating polyneuropathy (CIDP). PMID- 10689500 TI - IVIg in polymyositis and myasthenia gravis. PMID- 10689501 TI - High-dose intravenous immunoglobulin in multifocal motor neuropathy. PMID- 10689502 TI - Safety and tolerability of intravenous immunoglobulins. PMID- 10689503 TI - Monitoring the natural history and results of therapeutic intervention in diabetic neuropathy: clinical measures. PMID- 10689504 TI - Diabetic neuropathy--the utility of nerve biopsy. PMID- 10689505 TI - Nerve conduction study in diabetic polyneuropathy: multicenter analysis on intertrial variability. PMID- 10689506 TI - Autonomic tests in diabetic neuropathy. PMID- 10689507 TI - The Italian multicentre study on the prevalence of distal symmetric polyneuropathy: correlation between clinical variables and nerve conduction parameters. Italian Diabetic Neuropathy Committee. PMID- 10689508 TI - Utility of the skin biopsy method in studies of diabetic neuropathy. PMID- 10689509 TI - Clinical measures of disease activity in multiple sclerosis. PMID- 10689510 TI - Non-conventional MR techniques in monitoring MS activity and evolution. PMID- 10689511 TI - Immunological surrogate markers of disease activity in multiple sclerosis. PMID- 10689512 TI - Neurophysiological markers of relapse, remission and long-term recovery processes in MS. PMID- 10689513 TI - Monitoring temporal aspects of cortical information processing. PMID- 10689514 TI - Neural bases of time estimation: a PET and ERP study. PMID- 10689515 TI - [AINS (anesthesiology, intensive care, emergency medicine, pain therapy) 2000--a look to the future]. PMID- 10689516 TI - [Management and methodological approaches for the assessment of emotional states in anesthesiology]. AB - In anaesthesiology emotional states are of great importance. Reduction of anxiety and sedation in the preoperative preparation as well as stress reduction and the process of recovery are a challenge for anaesthetists as perioperative physicians. As emotions have different dimensions of manifestation like experience, expression, behaviour and somatic indicators, all these are needed to describe emotions sufficiently. In a multidimensional approach for the measure of emotional states, the different dimensions, their relationships and interactions are taken into account. The methodological approaches to registration of emotions in the anaesthesiological context are heterogeneous. In this summary the possibilities are differentiated by the source of information. Self-rating by the patient, rating by the observer, expression and behaviour and somatic indicators are taken into consideration. Analysis of the methods for the assessment of emotional states in anaesthesiological setting leads to the following recommendations: The most sensitive source of information is the patient. The rating scale used should be multidimensional and it should take specific as well as unspecific emotional aspects into account. As there are enough rating scales thoroughly developed and up to the demands of the classical test-theory, no ad hoc developed scales should be used. The rating of the emotional state should be supplemented by a rating of the physical state. The rating by the observer can be a valuable addition. The agreement between observers and the reliability of the method must be guaranteed. At presence there is no alternative in clinical practice to simple autonomic parameters such as blood pressure and heart rate as somatic indicators of emotion. Still it is important to consider the normal values for the individual patient. It is necessary to develop and to evaluate simple methods to register characteristics of expression in clinical context. PMID- 10689517 TI - [Target controlled infusion (TCI)--status and clinical perspectives]. AB - The technique of target controlled infusion (TCI) has influenced the development of intravenous anaesthesia substantially and opens the possibility of many new and exciting applications in peri-operative anaesthetic care. The launch of "Diprifusor" as the first commercially available TCI system for propofol was the cornerstone of a successful research period within the last decade, which evaluated the pharmacokinetic foundations of computer assisted intravenous drug delivery. We are now in a period where TCI technology is becoming a part of routine anaesthesia technique for the practitioner rather than a research tool for specialists and enthusiasts. This review gives an update on the rational pharmacokinetic basis of TCI development, the preliminary clinical experience with the new technique, the performance and accuracy of TCI devices and potential technical pitfalls in clinical routine. Besides clinical application in anaesthesia with "Diprifusor" TCI, target controlled systems are expected to play a significant role as research tools in the evaluation of drug interactions in anaesthesia and in the development of novel control techniques for the administration of sedative and analgesic drugs in the peri-operative period. PMID- 10689518 TI - [Direct autotransfusion systems deliver blood of inadequate quality]. AB - OBJECTIVE: Systems for direct retransfusion of blood salvaged from the surgical field and in drainage systems (direct autotransfusion) have been in use for many years. The quality of the blood obtained with such systems, however, has not been systemically assessed in a comparative manner. The aim of our study was the analysis of the quality of the blood, obtained with three commercially available direct autotransfusion systems (drainage systems with filters). METHODS: With ethics committee approval and informed consent, 30 patients receiving knee arthroplasty were randomly allocated to three groups. Each group of 10 patients received treatment with one drainage system (Consta Vac, Solcotrans, Haem-o Trans). In the salvaged blood, we measured cellular elements, variables of coagulation and fibrinolysis, complement activation and cellular elements, both before and after passage of the autotransfusion system. RESULTS: Analysis revealed a low haematokrit (< 30%) and platelet count (< 80 Gpt/l). The salvaged blood proved uncoagulable and defibrinised with no measurable clotting and fibrinogen; clotting activity, fibrinolysis and complement reaction were grossly induced (TAT, PAP and FDP high, C3 low). The blood was contaminated with cellular debris reflected by concentration of enzymes usually confined to the intracellular space (LDH, elastase, beta-thromboglobulin). CONCLUSION: The systems/filters assessed in this study do not improve quality of blood drained from the surgical field. Retransfusion of such blood can not be recommended. PMID- 10689519 TI - [Practical realization of a patient-accompanying concept in anesthesia and intensive care]. AB - Our current concept of stationary workplaces results in an interruption in patient monitoring and treatment. Because transfers are invariably associated with a reduction or interruption in the monitoring and treatment chain, an endangerment to critically-ill patients, as well as a significant increase in the mortality rates, can result. DESIGN: In the new construction of the Cardiac Clinic, the previous concept of immobile anesthesia and intensive care workstations has been completely abandoned. The complete treatment workstation, including monitoring and fluid management, is set up on a bedside cart which accompanies the patients uninterruptedly--from anesthesia administration, to the operating room, to the ICU or recovery room, as well as during elective or emergency interventions outside the ICU. Transport times and complications from 995 transports (ASA III and IV) were analysed and compared with 880 transports with the conventional system. RESULTS: During all intrahospital transfers with the mobile workplace, there were no complications resulting from faulty operation or accidental adjustment of the perfusors, or from disconnecting the monitoring, , respiration-, or infusion lines. On the whole, there were fewer cases of circulatory instability during transport, since infusion treatment and medication could be administered without interruption. All hemodynamic parameters were recorded during transport, as were cardiac minute output and right- and left atrial filling pressures. The mobile workplace system allows for the shortest possible transport and exchange times--13.5 min, as compared to 42.5 min with the conventional system. The reconnection of monitoring equipment with zeroing, adjustment of the alarm limits, as well as exchanging perfusors and infusomats before and after transport is eliminated entirely. CONCLUSION: This mobile workplace, in which all components of the anesthesiological and intensive care workstations are integrated, guarantees the highest possible level of patient safety, since nothing has to be disconnected until the patient is transferred to a normal-care ward. In addition to the improved ergonomic design of the nurse's and doctor's workplace, substantial savings can also be made. PMID- 10689520 TI - ["Is the Wurzburger pain drip an alternative to i.v. PCA?": contra]. PMID- 10689521 TI - ["Is the Wurzberger pain drip an alternative to i.v. PCA?": pro]. PMID- 10689522 TI - [Principles and role of nucleic acid amplification and modern microbiological diagnosis]. AB - In the recent decade, molecular tests have provided tools for highly sensitive and specific, culture-independent detection of infectious agents in clinical specimens. The rapid development of new methods and among these mainly the prototype method "polymerase chain reaction" (PCR) result in improved diagnostic procedures. Since its original description, a lot of modifications and advancements of PCR and alternative systems for in vitro amplification of nucleic acids have been developed to meet various requirements for improved detection of both DNA and RNA, quantification of the target molecules, and transfer from basic clinical research into a routine technique for clinical laboratory diagnosis. Since the purposes for which nucleic acid amplification methods should be used in the diagnosis of infectious diseases are often still uncertain, an evaluation and careful examination of the criteria for correct application of these techniques is needed. This review focuses on the recent developments in amplification procedures as well as on the use of these methods in the laboratory diagnosis of infectious diseases. The methodological limitations, future needs and perspectives are addressed. PMID- 10689523 TI - [Intraoperative management of unexpected arterial hypotension during lumbar diskectomy]. AB - An anesthesiological management is difficult in surgical procedures in which the operative situs is not to be seen by the anesthesist. Therefore specific knowledge of the operative procedure and related hazards are mandatory for optimal anesthesiological care. By describing the anesthesiologic proceduces in two lumbar discectomys, the specific problems in recognition and treatment of severe injuries of retroperitoneal vascular structures are explained. In addition, the differential diagnosis of intraoperative arterial hypotension is described. The communication between all disciplines involved is mandatory, especially in cases of severe complications. The management of such complications can only be solved through in-time interdisciplinary cooperation of all involved disciplines. PMID- 10689524 TI - Why history is important for thoracic surgeons. AB - There are numerous examples of lessons to be learned from acquaintance with surgical history. Notwithstanding these considerations--the admonition to read and think about history, the lessons learned from technical misadventures, and the need to add humanistic practices to our scientific endeavors--the real reward from our study of medical history lies in the pure job of being educated in one more way. This implies understanding our contemporary position in the unrolling course of medical history: from remote history through the enlightenment after the reawakening from the dark ages, to the surgical spurt in the latter half of the nineteenth century, and onward through the dramatic advances of our passing millennium. PMID- 10689526 TI - The history of surgery of empyema, thoracoplasty, Eloesser flap, and muscle flap transposition. AB - This article discusses the surgical history of empyema, thoracoplasty, the Eloesser flap, and muscle flap transfer. Little has changed in the 2000 years since the treatment of empyema was originally described by Hippocrates. The basic concepts of drainage of the infected empyema and obliteration of the space by allowing the lung to come up to the chest wall, taking the chest wall down to the lung, or by using muscle flaps or antibiotic solution remain the stabilizing forces in the treatment of postresection or postinfectious empyemas. PMID- 10689525 TI - The history of thoracic surgical instruments and instrumentation. AB - Thoracic surgical practice has evolved from the innovations of its pioneers. Beginning with the stethoscope discovered by Laennec with his system of auscultation, to the tools we use in the dissection and control of the hilum of the lung for resection, our practice of thoracic surgery has been entwined with the development of instruments and instrumentation. The development of strategies to prevent death from the open pneumothorax began with manual control of the mediastinum and progressed through differential pressure to, finally, the technique of intubation and the methods of positive-pressure and insufflation anesthesia. The instruments we place in our hands are not enough to define our art. Entry into the chest would not be possible without the use of rib retractors, rib shears, and even periosteal elevators. Finally, to the present day of minimally invasive techniques and the application of thoracoscopy for therapeutic purposes, we find the efforts of our predecessors well developed. For the progression from the fear of the open pneumothorax to the present-day state of the ease of thoracotomy for lung resection we are indebted to those who gave so much of their time and, for some, their lives to death from tuberculosis, to allow the advancement of our practice of surgery. These great people should be remembered not only for their acceptance of novel ideas but also, more importantly, for their lack of fear of testing them. PMID- 10689527 TI - History of surgery for penetrating chest trauma. AB - The military surgery experience of the past several centuries has been an important determinant of the evolution of the clinical management of penetrating thoracic trauma. The major management problems fall into two main categories: acute, life-threatening conditions such as open pneumothorax and exsanguinating hemorrhage, and chronic conditions such as clotted hemothorax, empyema, and fibrothorax. Better treatment and prevention of the latter has greatly reduced morbidity. Although hospital mortality has fallen by a factor of ten since the middle of the nineteenth century, the total mortality caused by penetrating thoracic trauma has undergone less change. PMID- 10689528 TI - The history of ventilation in the evolution of thoracic surgery. AB - The intrusion into the pleural space by surgeons was hindered for several hundred years by the realization that there were major pathophysiological alterations in ventilation and in circulation. The nature of this abnormality, although described very early on in history, went unrecognized until the end of the nineteenth century. The performance of thoracic surgery prior to that time and the development of different modes of ventilatory support are testimony to the intuition and inventiveness of the surgeons of that day. It is hard for the modern thoracic surgeon to fully comprehend the challenges that faced the early surgeon back when there was no such thing as positive pressure ventilation or unilateral lung ventilation. This article traces the origins of ventilation in man and their application to the development of thoracic surgery. PMID- 10689529 TI - The evolution of the surgical treatment of lung cancer. AB - The evolution of surgery for lung cancer is a story of discovery and innovation. From the fortuitous lung resections of the fifteenth century to the sophisticated operations of the twentieth century, surgeons have pursued the goal of bringing technology and science to bear on the effort to cure lung malignancy. Intrathoracic operations could not have developed without the advent of modern anesthesia, described in detail in another section of this issue. Great courage and insight were the hallmarks of those who first realized that surgical removal of primary lung cancer could become a reality and who pursued this goal in the face of discouraging results. The surgeons involved have worn many hats as experimentalists, physiologists, anesthetists, and biologists to bring all their knowledge and experience to bear on the surgical treatment of this disease. It is not possible in a brief review to identify the many physicians and scientists who contributed to the evolution of this treatment, but some of their stories have been included to illustrate the ideas involving major events over the past seven decades. PMID- 10689530 TI - The history of lobectomy and segmentectomy including sleeve resection. AB - The evolution of pulmonary lobectomy during the past six centuries is presented. The anesthetic problems of an open thorax, the absence of antibiotics, and the absence of radiology initially complicated thoracic surgery. Surgical pioneers differed strongly on the best techniques for lobectomy. Concurrence in their opinions evolved by the formation of a specialty group, meetings, and journals. The recent techniques of segmentectomy and sleeve resection provide further refinement of pulmonary resections. PMID- 10689531 TI - History of resectional surgery for tuberculosis and other mycobacterial infections. AB - Resectional surgery for tuberculosis became increasingly common in the 1940s; however, thoracoplasty remained the most popular treatment of choice until the introduction of effective antituberculosis agents. With the development of rifampin in 1966, surgery was seldom needed except for the occasional massive hemoptysis, bronchial stenosis bronchopleural fistula, or to rule out cancer. With the rise of MDR-TB and the increasing MOTT infections requiring surgery, resectional procedures are again being needed in the treatment of mycobacterial disease. PMID- 10689532 TI - History of esophageal surgery for benign disease. AB - The article examines the history of esophageal surgery for benign disease, looking at such surgeries as the first esophagotomy for a foreign body performed in 1738 and the first esophageal replacement of the esophagus in 1894. Various diseases are discussed, including stricture of the esophagus, hiatal hernia and gastroesophageal reflux, diverticula of the esophagus, and motility disorders such as achalasia. PMID- 10689533 TI - The history of surgery for carcinoma of the esophagus. AB - Carcinoma of the esophagus is a highly lethal disease in which surgical resection is part of every treatment regimen carried out with curative intent. The development of surgical resection of the esophagus for carcinoma has been a long and tortuous one. Its evolution depended not only on a thorough knowledge of surgical anatomy and technique, but also on important developments in endoscopy, radiology, anesthesia, nutrition, pulmonary physiology, and intensive care. PMID- 10689534 TI - The history of surgery of the thymus gland. AB - The history of surgery of the thymus gland is presented through highlights and landmark publications ranging from prehistoric times to the present day. The evolution of surgical techniques includes transcervical and transthoracic thymectomy along with other techniques such as current thoracoscopic VATS procedures. The significance of myasthenia gravis, autoimmunity, thymic oncology, adjuvant developments, and speculation on the future is discussed. PMID- 10689535 TI - Historical review of blunt injury to the thoracic aorta. AB - This article addresses the history of blunt injury to the aorta by reviewing the epidemiology, military accounts, ancient history, and recent history during the last five decades of the twentieth century. Although they are a bit arbitrary and overlap somewhat, significant groupings regarding blunt injury to the thoracic aorta have occurred in ten-year blocks. It is important that any surgeon not be locked into a previous time period but be continually knowledgeable of ever changing approaches. These approaches should always be based on the best evidence based information available. PMID- 10689536 TI - The history of surgery for thoracic outlet syndrome. AB - The history of surgery for thoracic outlet syndrome (TOS) is examined, ranging from the earliest recorded reference of TOS in the anatomic recognition of cervical ribs by Galen and Vesalius, to the current research on the diagnosis and treatment of TOS. The author discusses various cases that helped advance the use of surgery in treating TOS, tracing the years of progress that led to the present day understanding of the disease. PMID- 10689537 TI - The history of lung transplantation. AB - Experimental lung transplantation began in the former Soviet Union in 1946. Experiments in the 1950s sought to understand the physiology and to master the technique of reimplantation. Study of the immunology of allografting in the 1960s lead to the first lung transplantation in a human in 1963 in the United States. Palliative lung transplantation was first reported from Canada in 1986 after the advent of effective and tolerable immunosuppressive regimens. Lung transplantation in centers of excellence is now an effective palliative method for more than 80% of carefully selected recipients. Widespread transplantation to treat end stage pulmonary dysfunction is impossible because of too few organ donors and imperfect management of rejection and infection. PMID- 10689538 TI - Thoracic surgery training at the University of Michigan. AB - The process of thoracic surgery resident education is and always has been taken very seriously at the University of Michigan, where it is regarded as a key mission of the faculty. More attention is paid to the details of providing a supportive educational environment using established principles of education, curriculum planning, evaluation, and feedback. Resident education is the order of business today and, although challenged by the demands of managed care and cost containment, it remains among the most important priorities at the University of Michigan. PMID- 10689539 TI - The history of mediastinal teratoma. AB - The first discovery in 1823 of what has become known as mediastinal teratoma is discussed. The uniqueness of this tumor with its various spontaneous complications is described along with its early and often inept surgical therapies. This article highlights the development of the surgical treatment of mediastinal teratoma that has matured synchronously with the understanding of the physiology of the chest and the evolution of modern surgery. PMID- 10689540 TI - A diagnostic approach to musculoskeletal pain. AB - Musculoskeletal pain or inflammation is one of the most common causes of primary care office visits. Musculoskeletal disorders exact a high toll in distress, disability, and direct health care costs. Given the wide range of disorders that may cause or contribute to musculoskeletal symptoms, differential diagnosis is challenging and a systematic approach is necessary. Patient history is the single most valuable source of diagnostic information, followed by a careful physical examination. The history also suggests which laboratory tests and imaging studies, if any, are indicated. The chronology, duration, and pattern of pain distribution offer clues to establishing an accurate diagnosis, along with evidence of other organ system involvement or underlying disease. Helpful distinctions are those between articular and nonarticular pain, between monarthritis and multiple joint involvement, and between inflammatory and noninflammatory conditions. PMID- 10689541 TI - The role of the laboratory in the evaluation of rheumatic diseases. AB - This article presents information on the role of immunologic laboratory tests in evaluating patients with rheumatic disease. The focus is on 3 commonly used tests: antinuclear antibodies, rheumatoid factor, and the erythrocyte sedimentation rate. Background data on various statistical principles that are key to interpretation of these tests are also discussed. The goal is to emphasize that improper use and interpretation of such test results can lead to incorrect diagnosis and unnecessary therapy, potentially putting the patient at risk. Thus, these tests should be ordered in the context of other clinical information that provides the practitioner with an accurate estimate of disease probability. PMID- 10689542 TI - Osteoarthritis: a review. AB - In 1994, the Centers for Disease Control and Prevention reported that by the year 2020, arthritis will have the largest increase in numbers of new patients of any disease in the United States. The term arthritis refers to many diseases, the most common of which is osteoarthritis (OA). OA affects at least 16 million Americans, most of whom are older than 60 years. The disease is usually defined using radiologic criteria. More than 80% of people older than 75 years are symptomatic of OA. Considering cost of diagnosis, therapy (nonpharmacologic, pharmacologic, and surgical), side effects of therapy, and lost productivity, it is one of the more expensive and debilitating diseases in the United States. Given the large numbers of patients and the expense of the disease, it is not surprising that the diagnosis and care of patients with OA have come under scrutiny. The following article will provide some background on the disease and discuss management approaches that view the patient as a whole. PMID- 10689543 TI - Recent advances in the treatment of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) affects about 0.5% to 1% of the population worldwide. Because there is no cure for this disease, the goal of therapy is to control the underlying inflammatory process and maintain or improve function. This article reviews 4 new treatments for patients with RA: leflunomide, etanercept, infliximab, and the protein A immunoadsorption column with plasmapheresis therapy. PMID- 10689544 TI - Early detection research network in the US. PMID- 10689545 TI - Lack of association between body weight, bone mineral density and vitamin D receptor gene polymorphism in normal and osteoporotic women. AB - In an ethnically homogeneous population of women living in Tuscany, Italy, the relationships between age, body weight, bone mineral density and the vitamin D receptor (VDR) gene polymorphism were studied, with the objective of recognizing patients at risk for osteoporosis. In 275 women bone mineral density was measured by Dual Energy X-rays Absorptiometry (DEXA). In 50 of them the individual genetic pattern for VDR was evaluated by DNA extraction followed by PCR amplification of the VDR gene, and digestion with the restriction enzyme BsmI. Age and bone mineral density were inversely related (R2 = 0.298). Body weight was associated with bone mineral density (R2 = 0.059), but not with age. In osteoporotic women, mean (+/- SD) body weight was 59.9 +/- 6.5 Kg, lower than that recorded in non osteoporotic women (64.2 +/- 9.4 Kg), even though not significantly different (p = 0.18). No association was found between VDR gene polymorphism, bone density or body weight. The performance of anthropometric and genetic components appear to be poor, and, at least for the time being, bone mineral density measurement by means of MOC-DEXA represents the optimal method to detect women at risk for postmenopausal osteoporosis. PMID- 10689546 TI - Assessment of erythropoietin levels and some iron indices in chronic renal failure and liver cirrhosis patients. AB - This study was constructed to investigate the relationship between renal anaemia and erythropoietin (EPO) concentrations in chronic renal failure (CRF) patients and to evaluate the possible role of the liver. Serum EPO levels were measured in blood samples from 20 CRF patients on hemodialysis (HD), 20 liver cirrhosis (LC) patients, 20 patients having both CRF and LC and undergoing HD, and 20 normal control subjects. Blood cell counts, iron indices (iron, total iron-binding capacity (TIBC) and ferritin), renal function (blood urea nitrogen (BUN) and creatinine), hepatic function (ALT, AST, ALP and bilirubin) investigations were carried out for all the subjects enrolled in this study. CRF patients without LC had serum EPO concentration of 6.21 +/- 0.53 mU/ml (mean +/- SE), which was significantly higher than that in patients having both CRF and LC (4.32 +/- 0.52) (p < 0.01). Both groups showed significantly lower values than the controls (12.75 +/- 0.70) (p < 0.001). LC patients with intact kidneys had significantly higher EPO level (22.70 +/- 1.70) (p < 0.001). No correlation was found between EPO level and any of the hematologic or iron indices. PMID- 10689547 TI - Serum lipoprotein (a) levels in chronic renal failure and liver cirrhosis patients. Relationship with atherosclerosis. AB - This study was carried out to investigate the relationship between lipoprotein (a) levels and the development of atherosclerosis in chronic renal failure (CRF) patients with the possible role of the liver. Serum Lp (a) levels were measured in samples from 20 CRF patients on hemodialysis (HD), 20 liver cirrhosis (LC) patients, 20 patients having both CRF and LC and undergoing HD, and 20 normal control subjects. Renal function (blood urea nitrogen (BUN) and creatinine), hepatic function (transaminases (ALT and AST), alkaline phosphatase (ALP) and total bilirubin) investigations and serum cholesterol were carried out for all the subjects enrolled in this study. Serum Lp (a) concentration in CRF patients without LC was 87.25 +/- 6.17 mg/dl, which was significantly higher than all the investigated groups (P < 0.001). Lp (a) concentration in patients with both CRF and LC was 24.65 +/- 1.98 mg/dl, which was not significantly different from the controls, but was significantly higher than that in the subjects with LC only (P < 0.001) where the latter group had significantly low Lp (a) values (11.1 +/- 0.99) relative to all the other groups (P < 0.001). Lp (a) correlated positively with cholesterol in all groups except the LC subjects, but did not correlate with age, or renal function in both CRF groups. PMID- 10689548 TI - Transforming growth factor-beta and nitrates in epithelial ovarian cancer. AB - The role of transforming growth factor-beta (TGF-beta) and nitric oxide (NO) in ovarian neoplasia is still not clear. We studied the expression of TGF-beta by enzyme immunoassay, and nitrates (as a stable end product of NO) in 127 ovarian tissues (36 normal, 37 benign, and 54 malignant). Ploidy status and synthetic phase fraction (SPF) were also assessed by flow cytometry. Mean ranks of TGF beta, nitrate, and SPF were significant among different groups (X2 = 12.01, P = 0.0025, X2 = 67.42, P = 0.000, X2 = 9.06, P = 0.011 respectively). Nitrate mean ranks were significant among different FIGO stages of the disease (X2 = 17.6, P = 0.000). A significant correlation was shown between TGF-beta, and nitrate levels in all tissues (r = 0.24, P = 0.01), as well as in malignant tissues (r = 0.3, P = 0.026). Cutoff values were determined for both TGF-beta (290 pg/mg protein), and nitrates (310 nmole/mg non protein nitrogenous substances). At these cut offs, nitrates showed a sensitivity of 93% and 84% specificity for malignant versus normal cases, while TGF-beta had 76% sensitivity, and 82.4% specificity for poor versus good outcome. Patients with epithelial ovarian cancer were followed up for a total of 40 months. Survival analysis showed that patients with TGF-beta above the cut-off had worse prognosis (X2 = 12.69, P = 0.004). The present results suggest that malignant transformation of ovarian tissues is associated with increased TGF-beta and NO production. NO level is related to the development and progression of epithelial ovarian cancer, while high levels of TGF-beta could be of prognostic significance. PMID- 10689549 TI - Menstrual cycle dependent variability for serum tumor markers CEA, AFP, CA 19-9, CA 125 and CA 15-3 in healthy women. AB - Information on menstrual cycle dependent variation of tumor markers in healthy women is a subject of diagnostic efficiency and has an impact in elucidating the normal function of these markers. In this study midfollicular and midluteal concentrations of serum CEA, AFP, CA 19-9, CA 125, CA 15-3 and their relations with LH, FSH, prolactin, estradiol and progesterone were evaluated during ovulatory cycles in a group of 23 healthy female individuals. Samples were collected on the 7th and 21st day of the same menstrual cycle. Tumor marker and hormone concentrations were determined with chemiluminescence or electrochemiluminescence EIA methods. A significant phase-dependent difference was observed for CA 15-3, midluteal concentrations (mean +/- SEM; 26.33 +/- 1.56 U/ml) higher than the midfollicular (mean +/- SEM; 19.27 +/- 1.49 U/ml) concentrations (p < 0.001). But an obvious difference for other tumor markers investigated did not exist. Significant correlations of follicular and luteal CA 125 levels with body mass index of the subjects were observed (r:0.52, p < 0.05 and r:0.57, p < 0.005, respectively). CA 15-3 antigen is a product of the MUC-1 gene which is expressed in abundance by endometrial epithelial cells in the secretory phase of the menstrual cycle which may be the potential source of variability. The association of CA 125 levels with obesity suggests a possible role of adipose tissue in CA 125 metabolism. In conclusion our data suggest that in healthy women serum CA 15-3 levels are significantly elevated in the midluteal phase of the menstrual cycle compared to midfollicular phase. Therefore, consideration of menstrual cycle dependent variability for CA 15-3 appears indicated in interpretation of individual results. PMID- 10689550 TI - Novel, non-radioactive, simple and multiplex PCR-cRFLP methods for genotyping human SP-A and SP-D marker alleles. AB - We have previously identified an allele of the human SP-A2 gene that occurs with greater frequency in an RDS population [12]. Because of the importance of SP-A in normal lung function and its newly emerging role in innate host defense and regulation of inflammatory processes, we wish to better characterize genotypes of both SP-A1 and SP-A2 genes. It has been determined that SP-D shares similar roles in immune response. Therefore, in this report we 1) describe a novel, non radioactive PCR based-cRFLP method for genotyping both SP-A and SP-D; 2) describe two previously unpublished biallelic polymorphisms within the SP-D gene; 3) present the partial sequence of one new SP-A1 allele (6A14) and describe other new SP-A1 and SP-A2 alleles; and 4) describe additional methodologies for SP-A genotype assessment. The ability to more accurately and efficiently genotype samples from individuals with various pulmonary diseases will facilitate population and family based association studies. Genetic polymorphisms may be identified that partially explain individual disease susceptibility and/or treatment effectiveness. PMID- 10689552 TI - Placental growth, fetal growth and maternal RhE genotype. PMID- 10689551 TI - Lipid peroxidation and antioxidant status in human cervical carcinoma. AB - Reactive oxygen species (ROS), represented by superoxide, hydrogen peroxide and hydroxyl radicals, have been implicated in many diseases including cancer. ROS have been known to play an important role in the initiation and promotion of multistep carcinogenesis. The cellular antioxidants play a crucial role in protection against neoplastic disease. However, very little is known about the antioxidant defense in cervical carcinoma. This is addressed in the present study. Lipid peroxides, glutathione content and the activities of antioxidant enzymes, together with vitamin C and E content, were estimated in patients who had carcinoma of the cervix, and the values were compared with those of normal women. The results showed a remarkable reduction in the content of glutathione, vitamin E and C. Activities of glutathione peroxidase and superoxide dismutase were also reduced in cervical cancer compared to normal controls (P < 0.001). This reduction was more marked in late stages (III, IV) than in early stages (I, II) (P < 0.001). Glutathione was reduced more in poorly differentiated tumors (grade III) than in well and moderately differentiated ones (grade I, II) (P < 0.05). Levels of lipid peroxides were found to be significantly higher in malignant than in normal tissue samples and their levels were correlated with advanced clinical stage (P < 0.001). Our results suggest impaired antioxidant status in carcinoma of the cervix. This impairment is related to tumor progression. PMID- 10689553 TI - Ultrastructural changes in microvessel with age in the hippocampus of senescence accelerated mouse (SAM)-P/10. AB - Microvessels in the hippocampus of aged SAM-P/10 (14 months old) showed the following ultrastructural changes compared with those of young-mature controls (3 months old): (1) the majority of capillaries had lost the smooth contours typical of young cases; (2) the luminal surface of capillaries showed irregularity; (3) the endothelial cytoplasm was thicker; (4) vesicles appeared more frequently in the endothelium; (5) interendothelial tight junctions and basement membranes, however, seemed to show no significant abnormalities; (6) pericytes, especially those of arterioles and venules, contained many enlarged cytoplasmic inclusions with honeycomb-like vacuoles; (7) the area of glial perivascular end feet was greater. These morphological findings raise the possibility of impaired blood brain barrier function and microhemodynamic disturbances in aged SAM-P/10 hippocampus. PMID- 10689554 TI - Learning and self-regulation of slow cortical potentials in older adults. AB - Two groups of subjects, aged 20-28 and 50-64, respectively, matched for health status and verbal abilities, learned to control their slow cortical potentials (SCP) in a feedback paradigm by producing, on command, SCP shifts in either positive or negative direction. Both groups were able to differentiate significantly between the positivity task and the negativity task, with the differentiation score being only slightly (and not significantly) lower in older than in younger subjects. In all conditions, however, significantly more negative brain responses were obtained in older than in younger subjects. This effect was larger in the positivity task versus negativity task, and larger in trials without continuous SCP feedback versus trials with feedback. Additionally four learning tasks were carried out with all subjects. The older group demonstrated substantial performance deficits in two tasks with explicit learning (verbal and visual). In contrast, implicit learning (perceptual learning and skill acquisition) was not impaired with age. The results are at odds with the idea of general age-related learning deficit and concur with the hypothesis that only explicit, but not implicit, learning processes are compromised in older subjects. The pattern of consistently more negative SCP shifts produced by elderly subjects may indicate their impaired cortical inhibition. Another interpretation, which does not exclude the inhibitory deficit hypothesis but seems to better agree with other psychophysiological data, may be that older subjects have disturbance in the system controlling arousal and effort. PMID- 10689555 TI - Aging and negative priming: is ignored information inhibited or remembered? AB - We had younger and older adults complete two tasks that tested the attentional- and memory-based inhibition models of negative priming. One task violated May, Kane, & Hasher (1995, Psychological Bulletin, 118, 35-54) criteria for measuring just attentional inhibition, by including a repeated-target condition. The other task complied with these criteria and included a depth of processing manipulation, where participants selected prime targets based either on their letter-length (nonsemantic processing) or weight (semantic processing). On balance, results supported the memory model, because depth of processing clearly moderated younger adult negative priming, and older adults displayed negative priming only in the task satisfying the attentional-inhibition criteria (i.e., the depth of processing task). We conclude that memory factors moderate negative priming, and that May et al.'s criteria fail to predict when older adults will show the effect. PMID- 10689556 TI - Age effect in recall performance according to the levels of processing, elaboration, and retrieval cues. AB - The present study was conducted to investigate the incidence of several factors contributing to age-related memory decrement. Variables manipulated include quality (level of processing encoding conditions), the degree of effort and encoding quantitative elaboration (active/passive encoding conditions), and the influence of retrieval support (free-/cued recall conditions). In support of the environmental support hypothesis, middle-old and old subjects benefited more than young ones from cued recall in all the memory tests. Moreover, the results showed a differential (qualitative vs. quantitative) impairment of conceptual processing between the middle-old and the old-age groups. In the middle-olds, age differences were abolished by deep processing in old adults, age differences were attentuated only with deep and active processing associated with retrieval support. These gradual memory impairments are evaluated according to Mandler's model of memory (1979, In L. G. Nilsson [Ed.], Perspective in memory research. Hillsdale: Lawrence-Erlbaum), and the environmental support hypothesis is discussed in terms of the involvement of encoding and retrieval operations required by the memory task. PMID- 10689557 TI - Source monitoring and false recollection: a life span developmental perspective. AB - In a variation of Deese's (1959, Journal of Experimental Psychology, 58, 17-22) list-learning paradigm, 32 first-graders, 32 younger adults, and 24 older adults self-generated words that were semantically related to study items prior to recall. This manipulation increased false recollection for children and older adults, but not for younger adults. These data suggest that source-monitoring deficits underlie children's and older adults' illusory memories within the list learning format. The differential roles played by source monitoring versus declarative memory in the production of false memories are discussed from a life span developmental perspective. PMID- 10689558 TI - Perceived ability and level of education as predictors of traditional and practical adult problem solving. AB - Sixty adults (ages 19 to 80) were divided into three age groups. Each individual completed the Problem Solving Inventory (PSI), which assesses an individual's self-perception of problem-solving ability, prior to completing two types of problems, including concept-identification tasks and six everyday, practical problems. When the level of education across the younger, middle-aged, and older adults was controlled, older individuals perceived themselves as better problem solvers, and they were more confident in their problem-solving abilities than the other age groups. Additionally, these older adults with relatively high levels of education took more time to complete the concept-identification tasks but did not make more errors or need more choices to solve these tasks. Similarly, the older participants performed as well as either of the other age groups on the everyday, practical problems. PMID- 10689559 TI - Potential problems with the interpretation of hair analysis results. AB - Due to differences in hair growth rate depending on anatomical region, age, gender, ethnicity and interindividual variability, interpretation of parent drug or/and metabolite concentrations in hair is not easy. Furthermore, as drug incorporation mechanisms into hair matrix is not yet fully understood, it is rather difficult to extrapolate details on time and dose from hair segment analysis. If incorporation sources other than from bloodstream (skin secretions and/or external/environmental contamination) are considered, interpretation becomes even more complicated. For evaluating possible passive contamination, it is essential to consider specific identification of metabolites, use of metabolite-to-parent drug ratios, assays of decontamination washes and analysis of specimens collected from other body parts. Cosmetic hair treatment, natural and artificial hair colour, differences in hair structure and specificity of analytical methodology may represent other bias sources affecting concentrations of drugs in hair. A suitable cut-off level related to the LOD will allow correct identification of drugs or metabolites in hair. Regarding the performance of different hair testing laboratories, little information is available at this time to what extent test results are comparable and their interpretation is consistent. Frequency of drug consumption and time intervals between multiple consumption or lag time between consumption and appearance in the hair has not been fully investigated and needs further research. PMID- 10689560 TI - Statistical examination of hair color as a potential biasing factor in hair analysis. AB - We review eight different data sets in this paper for the purposes of assessing the possibility that reported color of hair can produce a systematic bias in the interpretation of hair assays. We review studies or data sets that include heroin and its metabolites, cocaine and its metabolites, MDMA and its analogs, and amphetamine and methamphetamine. The studies have utilized a variety of different degrees of color categorization, ranging from the simple dichotomy of brown and black, to a high of 12 categories. The mean number of categories reported approaches 6 (mean = 5.875). There are a total of 2791 data points in this analysis. We utilize two major statistical techniques for assessing significance; one-way analysis of variance, and Tukey's Honestly Significant Difference procedure. In circumstances were only dichotomous contrasts are possible, one-way analysis of variance is used. In contrasts involving three or more categorical groups, Tukey's procedure is used. In circumstances where the homogeneity of group variances is not sustained by the Levene statistic, we use the Tamahane procedure, allowing an assessment that assumes unequal variances. The analysis of this data fails to discern a significant color effect. We speculate that it may be that variance is large in many domains affecting analyte recovery from hair. In large groups these variations tend to regress towards a typical or mean value. Thus the data here show that while there are group or aggregate differences in these 'typical' values, they are not great when considered in relation to the within-group variations which exist for those values. It is our view that color may play a role in the accumulation of drugs in hair, however it is likely to account for only a very small part of the complex process of drug accumulation. PMID- 10689561 TI - Evidence for bias in hair testing and procedures to correct bias. AB - A number of in vitro experiments show that different hair samples incorporate differing amounts of drugs under identical conditions. Incorporation of cocaine and morphine tends to be correlated with race, in that the hair of African American females incorporates higher concentrations of cocaine than does the hair of Caucasian males or females. Extrapolation of these data into populations has been fraught with difficulties because the dosages of drugs and their use patterns are unknown. Cosmetic treatments and hygiene alter drug binding, which must be considered in comparing populations because cosmetic treatments are often group dependent. Four reasons are proposed that account for the uptake and retention of drugs by hair and that may differ among groups: (1) permeability and other characteristics of the hair due to genetic influences, (2) cosmetic hair treatments and hair care habits (which may be culturally influenced), (3) drug removal during personal hygiene, and (4) manner and route of drug administration which can affect passive exposure to residual drugs in the environment. The data supporting bias in hair testing are reviewed and methods are proposed that use either the uptake of dyes or the incorporation of drug homologs to reduce bias. PMID- 10689562 TI - Hair analysis for drugs of abuse. Hair color and race differentials or systematic differences in drug preferences? AB - There is currently a debate in the literature on chemical drug analysis concerning the contribution of biophysical attributes associated with specimens and specimen donors to assay outcome. In recent years this debate has focused on hair analysis, but has in the past also been raised in urinalysis interpretation. In this article we examine several aspects of that controversy. First, we present data regarding the effects of hair color on the distribution of positive hair testing results for three drug classes. We compare these results to negative hair samples from comparable donors. This data is derived from head hair from preemployment donors that was classified according to seven visual color categories. We determined the distribution of colors for hair samples devoid of any of three assayed drugs (amphetamines, cocaine, and cannabinoids). Subsequently, this distribution was compared with the distributions for hairs that had tested positive for amphetamines, cocaine or cannabinoids. We examined a total of 2000 randomly selected samples; 500 negative hair samples and 500 positive samples for each of three drugs: cannabinoids, cocaine, and amphetamine. We also evaluated ethnic/racial factors in relation to positive urinalyses for various ethnic/racial groups. We examined approximately 4000 urine specimens from two different groups, each constituting around 2000 specimens. In addition to ethnicity/race and urinalysis outcome, we also examined the relationship between the hair color distributions of urine donors and the corresponding urinalysis results for the three drug classes. We also compared them to drug-negative samples. Our summary impression is that the observed outcome patterns were largely consistent with differences in drug preferences among the various societal groups. There was little evidence of a pattern attributable to hair color bias alone or selective binding of drugs to hair of a particular color. Likewise, there was no discernible pattern associated with race or ethnicity that would lend support to a "race effect" in drug analysis. PMID- 10689563 TI - Influence of bleaching on stability of benzodiazepines in hair. AB - In order to study the influence of hair bleaching on benzodiazepines concentrations, hair was treated with a bleaching product (Poly Blonde, Schwarzkopf & Henkel) for 20 min. The treated hair specimen was obtained from a person who died after an overdose of several illicit drugs associated with benzodiazepines. Bleached and non bleached hair were washed (acetone and water), pulverised and then incubated for 2 h in a thioglycolic solution. In the extracts obtained by solid-phase extraction on C18 columns, the different drugs with the corresponding deuterated standards were derivatized and determined by GC-MS in a SIM mode. These results show that the concentrations of all the drug detected decreased in bleached hair in comparison with non treated hair. Whereas the diminution was less important for cocaine and benzoylecgonine (decrease of 24.6 and 36.4%, respectively), concentrations for codeine, 6-monoacetylmorphine and morphine decreased more significantly (decrease of 57.5, 88.6 and 67.4%, respectively) as well as those of diazepam, nordazepam and 7-aminoflunitrazepam (decrease of 39.7, 67.7 and 61.8%, respectively). The results in this study agree with those of other authors that bleaching affects the stability of cocaine and opiates incorporated in hair. These findings also point out that bleaching influences the stability of entrapped benzodiazepines in hair. Finally, these results reconfirm that it is very important to consider the cosmetic history of a hair sample in the interpretation of hair analysis results. PMID- 10689564 TI - The incorporation of dyes into hair as a model for drug binding. AB - The binding of charged substances from external aqueous media to hair has been investigated through the use of fluorescence microscopy. Eleven hair samples, reflecting various ethnic groups and cosmetic treatments, were tested. Rhodamine 6G, a cationic dye representative of drugs such as cocaine and opiates, showed incorporation throughout the hair of all samples except one. In contrast, fluorescein, an anionic dye representative of drugs such as THC carboxylic acid, was not readily incorporated. The incorporation of rhodamine 6G was faster for chemically 'straightened' and bleached African-American female hair than for untreated hair. Incorporation of rhodamine 6G followed a pH dependence, but an ionic strength dependence could not be established. These studies support three postulates: (1) electrostatic interactions explain the preferential binding of cationic drugs of abuse to hair; (2) the hair matrix, or the non-helical portion of hair, is accessible to external solutions and thus subject to contamination; and (3) cosmetic treatments may alter the helical portion of hair thereby increasing its accessibility to external contamination. PMID- 10689565 TI - Experimental designs in the optimisation of ultrasonic bath-acid-leaching procedures for the determination of trace elements in human hair samples by atomic absorption spectrometry. AB - Experimental designs were used for the optimisation of acid-leaching procedures assisted by ultrasonic energy for the extraction of Ca, Cu, Fe, Mg, Mn and Zn from human hair samples. A Plackett-Burman 2(7) x 3/32 design for seven factors ([HNO3], [HCl], [H2O2], acid/oxidant solution volume, exposure time to ultrasounds, temperature of the ultrasonic bath and hair particle size) was used in order to choose the variables affecting the acid-leaching process. The variables [HNO3], [HCl] and temperature of the ultrasonic bath were found to be the most important parameters for the acid-leaching procedure, and these variables were optimised by a response surface design (central composite design 2(3) + star) which involved 16 experiments. Optimum values in the 3.7-4.2 M range were found for [HNO3], while optimum values between 3.0 and 3.5 M were found for [HCl]. The optimum temperature of the ultrasonic bath was between 80 and 90 degrees C. An acid digestion induced by microwave energy (details given) was used to obtain the total metal concentration and also for comparative purposes. Ca, Cu, Fe, Mg and Zn were measured by flame atomic absorption spectrometry (FAAS) using a conventional air/acetylene flame, while Mn was determined by electrothermal atomic absorption spectrometry (ETAAS) under optimised conditions. Two different reference materials, IAEA-085 International Atomic Energy Agency, Monaco) and NIES No. 13 (National Institute for Environmental Studies, Japan), with certified metal contents for some of the elements investigated, were used in order to verify the accuracy of the methods. PMID- 10689566 TI - Hair analysis by using radioimmunoassay, high-performance liquid chromatography and capillary electrophoresis to investigate chronic exposure to heroin, cocaine and/or ecstasy in applicants for driving licences. AB - The present paper describes an integrated diagnostic strategy to check the physical fitness of subjects, formerly users of illicit drugs, to obtain a driving license, after having quit their addiction. According to the Italian law, applicants for a driving license with a history of drug abuse must give evidence to have quit this behaviour and to show no risk of relapse in the future. To prove this, at our institute, they undergo medical examination, hair analysis and a urinalysis program on eight seriate samples, collected over about 40 days. About 700 subjects per year are investigated with this strategy. The hair samples are screened for opiates (morphine), cocaine and ecstasy, the most abused illicit substances in our region, by using commercial radioimmunoassays adopting cut-off levels of 0.1 ng/mg. All positive samples and about 10% of negatives are confirmed by high-performance liquid chromatography. Further confirmation of results can be carried out by capillary electrophoresis (and/or GC/MS or MS/MS). In 1998, the prevalence of positives for morphine, cocaine and ecstasy was 4.8, 11.3 and 2.6%, respectively. In this year, for the first time, the percentage of hair samples positive for cocaine was greater than that for opiates. The results of this integrated diagnostic strategy are presented and discussed, with particular emphasis on the comparison between hair analysis on a single sample and seriate urinalyses (on eight samples). PMID- 10689567 TI - Use of headspace solid-phase microextraction (HS-SPME) in hair analysis for organic compounds. AB - Headspace solid phase microextraction (HS-SPME) has advantages of high purity of the extract, avoidance of organic solvents and simple technical manipulation and can be used in combination with gas chromatography-mass spectrometry (GC-MS) in the hair analysis of a number of drugs. HS-SPME coupled with the hydrolysis of the hair matrix by 4% sodium hydroxide in the presence of excess sodium sulphate and of a suitable internal standard proved to be a convenient one-step method for the measurement of many lipophilic basic drugs such as nicotine, amphetamine derivatives, local anaesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines. Detection limits were between 0.05 and 1.0 ng/mg. From spiked 10-mg hair samples absolute recoveries between 0.04 and 5.7% were found. These recoveries decreased considerably if larger sample amounts were used, perhaps due to increased drug solubility in the aqueous phase or to elevated viscosity in the presence of dissolved hair proteins. Because of the phenolic hydroxyl group a change of pH after alkaline hair digestion (by adding excess orthophosphoric acid) was necessary for the detection of delta 9-tetrahydrocannabinol (delta 9-THC), cannabinol (CBN) and cannabidiol (CBD) by HS-SPME. Nevertheless, the detection limits were such that only CBN could be detected in hair of a consumer. Clomethiazole, a compound hydrolysed in alkali, was measured by HS-SPME after extraction with aqueous buffer. The detection limit was 0.5 ng/mg. Cocaine could not be detected by HS-SPME. The application of HS-SPME to hair samples from several forensic and clinical cases is described. PMID- 10689568 TI - Selenium determination in mother and child's hair by electrothermal atomic absorption spectrometry. AB - A method for the selenium determination in a mother and her child's hair using palladium as a chemical modifier was optimized. The sample was digested with nitric acid and hydrogen peroxide and diluted to 5 ml. To achieve complete mineralization the samples were ashed at 1200 degrees C in the presence of palladium as a chemical modifier. The optimum atomization temperature was 1900 degrees C. The precision and accuracy of the method were studied using the reference material CRM 397. Results of calibration using aqueous standards and the standard addition method were compared. The method was applied to the selenium determination in 30 samples of the mother's and child's hair. The levels found were 0.54 +/- 0.34 microgram/g for mother's hair and 0.77 +/- 0.25 microgram/g for child's hair. PMID- 10689569 TI - Simultaneous hair testing for opiates, cocaine, and metabolites by GC-MS: a survey of applicants for driving licenses with a history of drug use. AB - A sensitive GC-MS method for the simultaneous determination of opiates, cocaine, and metabolites in hair at a cut-off level of 0.1 ng/mg was adopted to assess past exposure to these drugs in applicants for driving licenses with a history of drug use. The sampling protocol consisted of collection of one hair (sample A, 5 cm length) and one urine sample. When hair and urine (EMIT Syva, cut-off levels: 0.3 mg/l for opiates, 0.15 mg/l for cocaine, GC-MS confirmation of positives) were both positive or negative the protocol was concluded. In the other cases, the assessment of 'current exposure' to drugs was carried out, in order to avoid seriated random urinalysis, by collecting a second hair sample (sample B) 6 weeks later and analysing the proximal 1-cm segment. Out of the 214 'A' hair samples analyzed, 14 (6.5%) tested positive for morphine and/or 6-acetylmorphine (6AM), and 26 (12%) for cocaine and/or benzoylecgonine (BE), whereas none of the samples tested positive for both drugs. Levels between 0.1 and 1 ng/mg of the single analytes were found in eight out of the 14 morphine-6AM positives (57%) and in 18 out of the 26 cocaine-BE positives (69%). The time course of positive cases showed a progressive decrease of morphine-6AM positives and a corresponding increase of cocaine-BE positives within the study period September 1995-February 1999. No cases with positive urine and negative hair were observed. Among the 40 positive cases, seven (four and three for opiates and cocaine, respectively) were found to be 'currently exposed to drug', four by urinalysis (three and one) and three by analysis of the hair sample B (1 and 2). PMID- 10689570 TI - Tandem mass spectrometry: a helpful tool in hair analysis for the forensic expert. AB - The Bavarian State Bureau of Investigation in Munich has the exclusive responsibility for investigation of criminal acts. One considerable expertise is that of hair analysis. According to the legal system in Germany, there is a special interest when some clients' hair tested positive for illicit drugs. An accused with a lot of drugs in his hair will be treated as a supposed addict and will be guaranteed extenuating circumstances. The instrumentation used for hair analysis is a powerful analytical tool: a Varian 3400 gas chromatograph linked to a Finnigan Tandem-MS (TSQ 700). The methanol extraction method is used for the detection of illegal drugs and metabolites: amphetamine, methamphetamine, MDA, MDMA (ecstasy), MDE, MBDB, methadone, THC, EDDP (metabolite of methadone), cocaine, benzoylecgonine, cocaethylene, opiates (dihydrocodeine, codeine, heroin, 6-monoacetylmorphine, morphine, acetylcodeine). For the detection of 9-carboxy THC by negative chemical ionization the hair sample is hydrolyzed under alkaline conditions. Solid-phase extraction is used for clean-up. The LOQ for the determination of 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic-acid is 0.16 pg/mg hair. An unsurpassed combination for rendering an expert opinion based on hair analysis may be: a forensic expert using diligence and experience, coupled with the performance of a sophisticated analytical instrument. PMID- 10689571 TI - Analysis of LSD in human body fluids and hair samples applying ImmunElute columns. AB - Immunoaffinity extraction units (LSD ImmunElute) are commercially available for the analysis of lysergic acid diethylamide (LSD) in urine. The ImmunElute resin contains immobilized monoclonal antibodies to LSD. We applied the ImmunElute procedure to serum and also to human hair samples. For hair analysis the samples were first extracted with methanol under sonication. The extracts were then purified using the ImmunElute resin. LSD analysis was carried out with HPLC and fluorescence detection. The immunoaffinity extraction provides highly purified extracts for chromatographic analysis. The limit of detection (signal-to-noise ratio = 3) has been determined to be < 50 pg regardless of which sample material was used. The procedure was applied to authentic hair samples from drug abusers (n = 11). One of these samples tested positive with an amount of 110 pg LSD in 112 mg extracted hair corresponding to a concentration of 1 pg/mg. PMID- 10689572 TI - The use of supercritical fluid extraction for the determination of amphetamines in hair. AB - A laboratory study interested in the analysis of human hair for drugs-of-abuse was conducted to determine if drugs could be detected and quantified from hair. Supercritical fluid extraction (SFE) techniques followed by GC-MS analysis were applied to extract amphetamines from hair. The group of amphetamines included methylenedioxyamphetamine (MDA), methylenedioxymetamphetamine (MDMA), methylenedioxyethylamphetamine (MDEA) and internal standard mephentermine (MP). To validate information on amphetamine use in hair, powdered hair samples free from drugs were collected and soaked in a known amphetamine standard solution. Authentic fortified case hair samples taken from known drug users known to have consumed amphetamines were also analyzed for amphetamine. Results from this study show that amphetamine use can be detected in spiked and authentic fortified human hair using SFE techniques for qualitative and quantitative reproducible results. PMID- 10689573 TI - Are there possibilities for the detection of chronically elevated alcohol consumption by hair analysis? A report about the state of investigation. AB - The analysis of suitable ethanol markers in hair would be an advantageous tool for chronic alcohol abuse control because of the wide diagnostic window allowed by this specimen and the possibility of segmental investigation. Between the markers practically used or thoroughly investigated in blood or urine, ethylglucuronide, fatty acid ethylesters, phosphatidylethanol, acetaldehyde adducts to protein and 5-hydroxytryptophol can be regarded as possible candidates also in hair, but preliminary data were found in the literature only for ethylglucuronide and acetaldehyde modified proteins. By using headspace gas chromatography and headspace solid phase microextraction in combination with gas chromatography-mass spectrometry (SPME-GC/MS), in alkaline hydrolysates of hair it was possible to determine between 17 and 135 ng/mg of ethanol beside acetone and several other volatile compounds with slightly higher ethanol values for alcoholics than for social drinkers and teetotalers. A part of this is ethanol only absorbed in the hair matrix from the surrounding environment and consequently is not applicable as a diagnostic criterion. By extraction with aqueous buffer, methanol or a methanol/chloroform mixture and subsequent alkaline hydrolysis it was found that another part is generated from ethylesters, which are preferentially deposited in the lipid fraction of hair. In a specific search for ethylesters of 17 carboxylic acids by GC/MS-SIM in most cases ethyl 4 hydroxybenzoate (0.1 to 5.9 ng/mg, a preservative in hair cosmetics) and in four cases traces of indolylacetic acid ethylester were found. Furthermore, diethyl phthalate (a softening agent, present also in many cosmetic products) was identified in the hair of alcoholics as well as of children. As potential markers of alcohol intake, ethyl palmitate, ethyl stearate and ethyl oleate were detected in hair samples of alcoholics by headspace SPME-GC/MS of the chloroform/methanol extracts. PMID- 10689574 TI - Use of solid-phase microextraction (SPME) for the determination of methadone and EDDP in human hair by GC-MS. AB - Solid-phase microextraction (SPME) is a new extraction technique with many advantages: small sample volume, simplicity, quickness and solvent-free. It is mainly applied to environmental analysis, but is also useful for the extraction of drugs from biological samples. In this paper the use of SPME is proposed for the determination of methadone and its main metabolite EDDP in hair by GC-MS. The hair samples were washed, cut into 1-mm segments, and incubated with Pronase E for 12 h. A 100-micron polydimethylsiloxane (PDMS) film fibre was submerged for 30 min in a diluted solution of the hydrolysis liquid (1:4 with borax buffer) containing methadone-d3 and EDDP-d3 as internal standards. Once the microextraction was concluded the fibre was directly inserted into the CG injection port. Linearity was found for methadone and EDDP in the range studied, 1.0-50 ng/mg hair, with correlation coefficients higher than 0.99. Interassay relative standard deviation (R.S.D) was determined to be less than 13.30% for methadone and less than 8.94% for EDDP, at 3.0 and 30.0 ng/mg. Analytical recoveries were close to 100% for both compounds on spiked samples. The method was applied to the analysis of real hair samples from eight patients of a methadone maintenance programme. The concentration of methadone in hair ranged from 2.45 to 78.10 ng/mg, and for EDDP from 0.98 to 7.76 ng/mg of hair. PMID- 10689575 TI - Incorporation of propyphenazone in beard hair of a migraine patient. AB - The incorporation of propyphenazone in beard hair after consumption of this substance present in the analgesic Migraine-Kranit (Codali) was investigated. Because of a migraine attack a volunteer took four tablets of Migraine-Kranit (one tablet contains 150 mg propyphenazone) the first day and two tablets the second day. Shaved beard hair was collected 48, 72, 96 and 120 h after the first consumption of the analgesic drug. These hair specimens were washed (acetone and water), pulverized and then incubated during 2 h in a thioglycolic solution. After solid-phase extraction on C18 columns, propyphenazone was assayed in these extracts by GC/MS operating in selected ion monitoring mode (m/z 230, 215). Diazepam-d5 was used as an internal standard. In hair specimen 1 (48 h after consumption) the highest concentration was found (170 pg/mg hair). In hair specimen 2 (72 h) and 3 (96 h) the concentration were significantly lower (44 and 18 pg/mg, respectively). After 120 h no propyphenazone could be detected (limit of detection: 5 pg/mg hair). These results show that propyphenazone was already in beard hear 2 days after consumption, whereas no more presence could be shown after 120 h. As the time period of 2 days is too short to allow entrapment into the hair matrix from bloodstream and growing of hair out of the follicle, our results suggest that incorporation of propyphenazone may be mainly due to excretion in sweat and subsequent incorporation into the hair. PMID- 10689576 TI - Detection of THCCOOH in hair by MSD-NCI after HPLC clean-up. AB - Regular consumption of cannabis can easily be detected by examination of hair for tetrahydrocannabinol, cannabinol, and cannabidiol. Although several studies have demonstrated that after contamination with smoke or treatment with THC containing shampoos THC is not detectable, or only in small traces, the detection of 11-nor 9-carboxy-delta 9-tetrahydrocannabinol (THCCOOH) should be offered to prove the consumption and metabolisation of THC. Up to now this confirmation was only available using tandem MS techniques combined with negative chemical ionisation. A new method using a normal quadrupole GC/MS is described. The lack of expensive instruments has to be paid for by a costly and time consuming extraction and clean-up. After the sample has been digested by 2 M NaOH at 95 degrees C and the neutralised liquid has been extracted with a mixture of n-hexane and ethyl acetate the dried residue is reconstituted in acetonitrile-methanol-0.01 M sulfuric acid (49:21:30, v/v/v) and the cannabinoids separated by HPLC. Each fraction is collected over 1 min. Another extraction with n-hexane-ethyl acetate is followed by evaporation, derivatisation, and GC/MS determination. The calibration with THCCOOH spiked hair led to a LOD of 0.3 pg/mg and a LOQ of 1.1 pg/mg. PMID- 10689577 TI - Hair analysis by immunological methods from the beginning to 2000. AB - Immunoassays for hair testing must satisfy three requirements: (1) They must have cross-reactivity with parent drug and lipophilic metabolites actually found in hair (2) they must not experience interference from the dissolved hair matrix and (3) they must be titered for cutoffs appropriate to the drug concentrations found in hair. Because the analytes found in hair after drug use are generally the parent drug or its lipophilic metabolites, immunoassays developed and intended for urine testing are not suitable for hair. Immunoassays whose antibodies are bound to a solid support, such as coated-tube radioimmunoassay or coated-plate ELISA tests, experience less matrix interference than those which use other means of separation of bound and free fractions. Homogenous assays are not suitable for hair testing because the hair matrix frequently interferes in the detection of the signal. Historically radioimmunoassays for drugs of abuse were first used for detecting drugs in hair. Currently ELISAs and coated-plate 96 well microplate EIAs are employed for screening hair digests or extracts for drugs. The optimum cutoffs for immunoassays for drugs in hair should be chosen based on the analyte concentration which produces the fewest false positive or false negative results when applied to tests of hair from known users and non-users of drugs. A hair immunoassay test at these cutoffs should have a sensitivity and specificity of better than 90%. The predictive value of the test will depend on the prevalence of drug use in the tested population. Cutoffs or decision thresholds for immunoassays used for screening for drugs should not be at the limit of detection of the assay because that produces a very large incidence of false positives. Because immunoassays are ligand-binding assays, they have a short range of linearity with low precision at both ends of the range. In the future, immunoassays will continue to be used for screening hair and other matrices for drugs of abuse because they provide rapid, inexpensive automated procedures for separating negative specimens from those which are suspected of containing drugs. For forensic purposes, all positive results must be confirmed by an independent analysis using a procedure based on a different property of the analyte. An immunoassay test should not be confirmed by a second immunoassay test but by a chromatographic test performed on a different dissolved or extracted aliquot of the original specimen. PMID- 10689578 TI - Analysis of drugs of abuse in hair by automated solid-phase extraction, GC/EI/MS and GC ion trap/CI/MS. AB - In our laboratory, analysis of human hair for the detection of drugs of abuse was first performed in 1995. Initially, requests for hair analysis were few, and it is only since 1997 that these analyses have become routine. As demand grew, we developed an automatic solid-phase extraction method; the use of a robot ASPEC allowed us to drop certain fastidious manipulations, and to treat a large number of samples at a time. This method is described, along with analysis by gas chromatography-mass spectrometry (GC/MS) in selected ion monitoring mode (SIM), for the following drugs: codeine, 6-monoacetylmorphine (6-MAM), morphine, cocaine, methadone, ecstasy (MDMA) and Eve (MDE). This requires prior derivatization with propionic anhydride. The different validation parameters, linearity, repeatability, recovery and detection limits are described, as well as the application of this method to some real cases. Analysis of these cases is also performed by an ion trap GC/MS in chemical ionization mode (GC/IT/CI/MS) in order to demonstrate the usefulness of this technique as a complement to routine analysis. Analysis by GC/IT/CI/MS indeed avoids the risk of false-positive results by the identification of metabolites. PMID- 10689579 TI - Evaluation of cocaine, amphetamines and cannabis use in university students through hair analysis: preliminary results. AB - The evaluation of drug abuse in a defined population was performed through toxicological hair analysis. Hair samples from university students ranging from 18 to 25 years of age were anonymously collected and screened for cocaine, amphetamines and cannabinoids by radioimmunoassay (RIA). Positive results (cut off values adopted were 2 ng/mg for cocaine and amphetamines and 0.5 ng/mg for cannabinoids) were confirmed by GC/MS. Preliminary results showed 19% of positive results for cocaine on 200 samples analysed. No confirmed positive results were obtained for amphetamine analysis. RIA technique demonstrated its unsuitability for cannabinoids preliminary screening on hair, giving a high percent of false positive results. PMID- 10689580 TI - Clinical applications of hair testing for drugs of abuse--the Canadian experience. AB - During the last 2 decades there has been a substantial increase in illicit drug consumption in North America. It has been repeatedly shown that the personal history of drug use is far from being accurate. Fearing legal consequences and embarrassment of admitted illicit substance use, most users tend to deny or, to under-report illicit drug consumption. These facts have stressed an urgent need for a biological marker which does not lose its sensitivity within a few days after the end of exposure and which may yield a cumulative reflection of long term exposure to illicit drugs. Hair analysis has emerged as such a marker. A variety of illicit and medicinal compounds have been shown to be incorporated into hair including trace metals, barbiturates, amphetamines, opiates, phencyclidine, cocaine, nicotine and cannabis. Hair analysis for drugs of abuse provides long-term information on an individual's drug use; its window of detection is limited only by the length of the hair and typically, ranges from a week to several months. After establishing and validating several hair tests during the last decade, we have analyzed over 1000 hair samples for different drugs of abuse. We used RIA for screening and GC-MS for confirmation of positive results. The aim of this report is to illustrate the diagnostic usefulness of hair testing in different age groups (newborns, children, adults) and circumstances: (criminal cases, athletes, child custody cases, etc.). PMID- 10689581 TI - Clozapine dose-concentration relationships in plasma, hair and sweat specimens of schizophrenic patients. AB - The aim of the present study was to establish an analytical method for the determination of clozapine in sweat and to determine whether the clozapine level in hair and sweat were correlated to the daily dose of clozapine delivered to patients. Twenty-six subjects treated with clozapine at 200-700 mg/day for refractory psychosis were included in the study. Clozapine was determined in plasma by liquid chromatography coupled to a diode array detection system, after extraction with an organic solvent at pH 9.5. Clozapine was extracted from hair and sweat patches specimens by incubation in methanol overnight at 40 degrees C. The residues were analyzed by gas chromatography coupled to mass spectrometry in the electronic impact mode of detection. It was possible to determine clozapine in concentrations ranging from 30 to 1016 ng/ml in plasma (n = 22), from 0.17 to 34.24 ng/mg in hair (n = 23) and from 49 to 5609 ng/patch in sweat (n = 20). Preliminary results suggest a lack of correlation between daily regimen of clozapine and plasma levels of the drug. Therefore, a better dose-concentration relationship was observed in our study between daily dose and hair concentration (r = 0.542, P < 7%) or between daily dose and sweat concentration (r = 0.589, P < 6%), but with wide variations for patients at the same posology. However, the idea of using quantitative drug measurements in hair or sweat to ascertain whether a patient has taken his treatment exactly as prescribed will remain inapplicable. PMID- 10689582 TI - Hair analysis for driving licence in cocaine and heroin users. An epidemiological study. AB - Diagnosis of drug exposure is strongly supported by analysis of hair samples. In the province of Brescia, Italy, for regranting driving license to drug addicts or occasional abusers, a control programme was adopted including analysis of illicit drugs in two hair segments (0-3 and 3-6 cm) and in urine. From January 1998 to April 1999, upon request of the Local Medical Commission, 697 hair samples were tested in our laboratory. One hundred and eighty subjects resulted positive in hair for one or two of the controlled drug classes (73.3% for cocaine, 10% for opiates, 16.7% for both). Positive subjects were classified by residence, age, sex and license category. Seventy-two subjects were called back after 6-12 months and submitted to a second hair and urine analysis: in 34 cases the result of the first analysis was confirmed (19 negatives, 15 positives for one or both drug classes). Another 37 cases tested positive at the first control and negative at the second, suggesting the hypothesis that a strict control may have a significant deterrent function. The high percentage of negative results at the second control may be explained by the prevalence of cocaine users in the examined population. Our results allow us to conclude that the strict application of control rules lead to a decrease of social risk behaviours. PMID- 10689583 TI - Is there a place for hair analysis in doping controls? AB - The actual antidoping control rules applied in sports (as established by the International Olympic Committee and the International Sport Federations) state that a positive case is chemically established by the unequivocal detection of a forbidden parent molecule and/or any of its metabolite(s) in urine, no matter the amounts which were administered and when the drug was taken. Screening is accomplished most of the time by using GC-MS procedures. These have been optimized to detect most if not all of the forbidden compounds which are put on a list. Recently, attempts have been made on scalp hair to demonstrate the value of this matrix as a possible means for differentiating between therapeutic use and doping abuse. In particular, GC-mass selective detector and GC-high resolution MS were successfully applied to treated animals and body-builders for anabolic agents (steroids and beta-2-agonists) at high sensitivity detection (low ng/g level). Naturally occurring molecules, like testosterone and its metabolites, could also be differentiated from their synthetic counterparts. Positive cases are more often challenged in courts and retrospectivity in time of the drug(s) intake is becoming an important issue for evaluating the responsibility of the person. This is can be based on hair analyses if the drugs have been taken at regular intervals. Stimulants and narcotics are often used in sports like drug of abuse in the ordinary social contexts. On the other hand, anabolic agents, when taken to improve the physical performances, follow complex regimens with the mixing of various formulas and dosages. Scalp hair references ranges for these as well as for endogenous substances still wait to be established statistically for competing, well-trained athletes. The incorporation rate into blond or gray hair is poorer than that of dark colored hair raising the question of individuals equality against the controls, a very important matter of concern for the sport's governing bodies. The frequency of hair cutting and short hair cuts necessary to gain speed in specific sports like swimming are other critical factors. On the other hands, irregular hair growth, associated with the washout effect through multiple washing and staining processes over expanded time intervals can cause concentrating or diluting effects. So far, a minority of prohibited substances could be detected in scalp hair with the sensitivity and specificity required in the context of the sport's activities. From the above, clear limitations of the usefulness of hair analysis in doping control analysis are obvious until a lot more data relevant to this particular field have been collected. PMID- 10689584 TI - Pharmacological criteria that can affect the detection of doping agents in hair. AB - When positive drug results are reported, a common interpretive question posed is whether or not it is possible to put a quantitative finding into context. A standard answer to this inquiry is that a positive hair testing result can be interpreted as meaning that the donor has chronically or repetitively used the drug identified in the hair, but that chronic or repetitive are not defined in the same way for all individuals. The Society of Hair Testing published on June 16, 1999, a consensus opinion on the use of hair in doping situations. However, although accepted in most courts of justice, hair analysis is not yet recognised by the International Olympic Committee. To be considered as a valid specimen for doping control, some issues still need to be addressed. The scientific community has demonstrated significant concern over the proper role that hair drug testing should serve in toxicological applications. Among the unanswered questions, five are of critical importance: (1) What is the minimal amount of drug detectable in hair after administration? (2) What is the relationship between the amount of the drug used and the concentration of the drug or its metabolites in hair? (3) What is the influence of hair color? (4) Is there any racial bias in hair testing? (5) What is the influence of cosmetic treatments? The present report documents scientific findings on these questions, with particular attention to the applications of hair in doping control. PMID- 10689585 TI - Analytical strategy for detecting doping agents in hair. AB - Lists of banned classes of doping agents are released by the International Olympic Committee, adopted by other sports authorities and updated regularly, including the substance classes stimulants, narcotics, diuretics, anabolic agents, peptide hormones, beta-blockers etc. There are different classes of restriction: anabolic and masking agents (anabolic steroids, diuretics etc.) are always banned for athletes regardless of their topical activity (training or competition) several substances are permitted with certain restrictions (caffeine below a cut-off value, or inhalation of some beta 2 agonists) beta-blockers are prohibited in competitions of certain sports disciplines the majority of the substances (stimulants, narcotics etc.) is prohibited during competitions, so that they do not have to be analysed in out-of-competition samples. A differentiation between training and competition period is impossible by means of hair analysis due to the uncertainty of (especially short-term) kinetic considerations related to hair growth. Therefore, the analytical identification of doping relevant substances in hair is not always a sufficient criterion for a doping offence and the identification of stimulants, beta-blockers etc. in hair would be entirely irrelevant. The most interesting target substances are certainly the anabolic agents, because their desired action (enhanced muscle strength) lasts longer than the excretion, leading to sophisticated procedures to circumvent positive analytical results in competition control. Besides the analysis of out-of-competition control samples, the long term detection of steroids in hair could provide complementary information. An analytical approach to the identification of exogenous steroids in hair requires consideration of the presence of many other steroids in the hair matrix interfering the analysis at trace levels, and of a limited chemical stability. The analysis of endogenous steroids in hair appears to be even more complicated, because the possibility of many biotransformation reactions from (into) other precursors (metabolites) has to be taken into account. Precursor substances of anabolic steroids (especially esters as application forms) are very promising analytical targets of hair analysis, because they can only be detected after an exogenous intake. The quantitative evaluation of active parent compounds like testosterone (which is actively involved in physiological processes of hair growth) in hair is still controversial. Clinical applications under reproducible conditions can be useful, but the biovariability of these parameters will probably prevent the definition of acceptable cut-off levels as a criterion of abuse. PMID- 10689586 TI - Hair analysis and detectability of single dose administration of androgenic steroid esters. AB - Detection of anabolic steroids in hair samples has been possible only in fatal cases or in cases of high-continuous dosages. In order to verify the possibility of detecting an acute administration, a sensitive and specific assay has been developed for the simultaneous determination of testosterone, nandrolone and some of their esters in hair. The analytes were extracted from finely cut hair with methanol-trifluoroacetic acid overnight. After the incubation, the mixture was evaporated to dryness, redissolved and extracted with hexane. The dried organic layer was silanised and analysed by GC-MS and GC-MS-MS. A sensitivity of at least 20 pg injected was obtained for all the analytes. In guinea pigs treated with a single intramuscular dose of 10 mg/kg nandrolone decanoate, neither nandrolone decanoate nor nandrolone were found in hair collected after 13 days, while both compounds were clearly detectable after four repeated doses (each dose every 3-4 days) of 20 mg/kg nandrolone decanoate. Neither nandrolone decanoate nor nandrolone could be detected in hair from a male healthy volunteer 1 month after treatment with 50 mg nandrolone decanoate, while his urine still tested highly positive for the main nandrolone metabolite (> 100 ng/ml). Testosterone esters could not be detected in hair of healthy subjects collected respectively 3, 2 and 1 month after a single intramuscular administration of 250 mg testosterone enanthate (five subjects), a single intramuscular coadministration of 25 mg testosterone propionate plus 110 mg testosterone enanthate (one subject), or a single oral administration of 120 mg testosterone undecanoate (three subjects). Otherwise, hair analysis revealed an increase of testosterone concentration corresponding to the period of treatment. Analysis of blood and urine samples confirmed the absorption of those compounds. At the sensitivity achieved by the present method, no detection of nandrolone, nandrolone decanoate nor testosteron esters in hair seems to be obvious after a single dose administration. PMID- 10689587 TI - Compared interest between hair analysis and urinalysis in doping controls. Results for amphetamines, corticosteroids and anabolic steroids in racing cyclists. AB - In France during a famous bicycle race, the newspapers documented the degree in which doping seemed to be supervised in some teams by managers and doctors. Use of anabolic steroids and other substances was officially banned in the mid seventies by sports authorities. This policy has been enforced through urine testing before competition. It is well known, however, that a latency period is all that is necessary to defeat these tests. Nevertheless, hair analysis could be a promising tool when testing for periods that are not accessible to urinalysis any more. We have developed different sensitive methods for testing hair for amphetamines, anabolic steroids and their esters and corticosteroids. For amphetamines, 50 mg of hair were digested with 1 M NaOH, extracted with ethyl acetate, derivatized with TFA and analyzed by gas chromatography positive chemical-ionization mass spectrometry. For corticosteroids, 50 mg of powdered hair were treated with methanol in an ultrasonic bath and subsequently purified using a C18 solid phase extraction column. Analysis was realized by high performance liquid chromatography coupled to electrospray-ionization tandem mass spectrometry. For anabolic steroids and their esters, 100 mg of powdered hair were treated with methanol in an ultrasonic bath for extraction of esters, then alkaline digested with 1 M NaOH for an optimum recovery of other drugs. The two liquid preparations were subsequently extracted with ethyl acetate, pooled, then finally highly purified using a twin solid phase extraction on aminopropyl and silica cartridges. Residue was derivatized with MSTFA prior to injection. Analysis was conducted by gas chromatography coupled to a triple quadrupole mass spectrometer. Thirty cyclists were sampled and tested both in hair and in urine. Amphetamine was detected 10 times in hair (out of 19 analyses) compared to 6 times in urine (out of 30 analyses). Corticosteroids were detected 5 times in hair (methylprednisolone 1 case, triamcinolone acetonide 3 cases and hydrocortisone acetate 1 case) in hair (out of 12 analyses) compared to 12 times (triamcinolone acetonide 10 cases and betamethasone 2 cases) in urine (out of 30 analyses). Anabolic steroids were detected twice (nandrolone 1 case, and testosterone undecanoate 1 case) in hair (out of 25 analyses) compared to none in urine (out of 30 analyses). PMID- 10689588 TI - Identification of ten corticosteroids in human hair by liquid chromatography ionspray mass spectrometry. AB - This paper describes a screening procedure based upon high-performance liquid chromatography-ionspray mass spectrometry for the identification of ten corticosteroids in human hair: triamcinolone, prednisolone, prednisone, methylprednisolone, cortisone, cortisol, beta- and dexamethasone, flumethasone and beclomethasone. Hair strands were washed in methylene chloride, pulverized in a ball mill and 50 mg of the powdered hair were incubated in 1 ml Soerensen buffer, pH 7.6 for 16 h at 40 degrees C, in presence of 50 ng cortisol-d3 used as internal standard. Purification of the incubation medium was achieved on SPE C18 Isolute extraction columns. The eluates were evaporated to dryness and resuspended in 30 microliters MeOH before analysis by HPLC-IS-MS in positive and negative modes of detection. The validation parameters were found satisfactory for a corticosteroid screening procedure. The correlation coefficient of the calibration curve ranged from 0.939 to 0.997, showing linearity between 0.1 and 10 ng/mg, excepted for beclomethasone which was between 0.2 and 10 ng/mg. Extraction recovery at 4 ng/mg ranged from 43.2 to 85.7%. Repeatability (CV values) at 4 ng/mg ranged from 6.1 to 17.5%. The limits of detection ranged from 0.03 to 0.17 ng/mg for a signal-to-noise ratio of 2. The detection of prednisone and beclomethasone in three hair specimens obtained from forensic and clinical cases have documented corticosteroids incorporation into human hair. PMID- 10689589 TI - The hair analysis proficiency testing program of the French Society of Analytical Toxicology. AB - In an effort to improve laboratories performing hair analysis in forensic cases, the French Society of Analytical Toxicology (S.F.T.A.) has implemented a proficiency testing program since 1992. Actually about 10 laboratories are participating. Each survey is dedicated to one analyte or one pharmacological class: opiates (6-monoacetylmorphine, morphine and codeine), cocaine and benzoylecgonine, tetrahydrocannabinol, buprenorphine and norbuprenorphine, beta blockers (metoprolol, atenolol), beta 2-agonists (salbutamol, clenbuterol). Animal hair was tested for clenbuterol. Prior to sending, hair samples were reduced to a powdered form, well mixed to ensure homogeneity, and then tested by GC/MS or HPLC/MS. Results confirm those obtained in a preliminary study on opiates and cocaine analysis in hair: a common analytical procedure has to be used by all the participants, including hydrolysis of hair. It is essential to work on authentic drug-positive hair samples and not on spiked samples. Participation at this program is free of charge and considered as an educational tool. Comparison of the results with those of other laboratories in Europe and USA shows that the analytical methods used during this program are in accordance with all the other procedures. PMID- 10689590 TI - Implications of recent findings in posttraumatic stress disorder and the role of pharmacotherapy. AB - Recent evidence suggests that an etiologic model of posttraumatic stress disorder (PTSD) must include both vulnerability factors (presumably related to dysregulation of stress responses and/or failure of normal restitutive mechanisms following trauma) and factors related to trauma severity. The fact that rates of PTSD increase with the severity of trauma suggests that normal adaptive mechanisms may become overwhelmed even in the absence of vulnerability factors. Consistent with this view, efforts to demarcate normative from disordered reactions to severe trauma, such as the new diagnosis of acute stress disorder, have had limited success. Debate over the moral and scientific implications of receiving a trauma-related diagnosis has further complicated the issue and perpetuated a false dichotomy concerning normative responses. The literature on clinical trials in PTSD is reviewed. The range of treatment responses, and the categorical breadth of compounds studied, requires interpretation before the literature as a whole can be understood. One of the many limitations of this new literature is the absence of treatment-outcomes research on individuals with the common comorbidity of substance abuse. The most recent findings with selective serotonin-reuptake inhibitors and related compounds indicate a more optimistic outlook for pharmacological treatment of PTSD than was suggested by earlier trials. Given these observations, investigators will hopefully be encouraged to pursue study and development of treatment models that include both pharmacological and psychosocial interventions. PMID- 10689591 TI - The efficacy of newer antidepressants in the treatment of chronic pain: a review of current literature. AB - Tricyclic antidepressants have been extensively studied and frequently used in the treatment of various chronic pain syndromes. Newer antidepressants, namely fluoxetine, sertraline, paroxetine, fluvoxamine, citalopram, trazodone, nefazodone, bupropion, mirtazapine, and venlafaxine, have also been considered for this indication, although they have been less extensively studied. This article reviews the available publications, including placebo-controlled trials, other outcome studies, and case reports, pertaining to the use of these medications for chronic pain. Although some of these newer compounds may be effective for specific types of pain, making generalizations regarding their use as analgesics is difficult, given the limitations of existing data. Additional observations based on the data are presented in the hope that they may help to guide further research and clinical use. PMID- 10689592 TI - Huntington's disease and its association with psychopathology. AB - We review several aspects of Huntington's disease (HD), with a special focus on the psychopathological manifestations often identified in patients with this disorder. We discuss the evidence for a higher-than-average prevalence of psychosis, depression, and obsessive-compulsive disorder (OCD) in individuals with HD or at risk for the illness and analyze the possible significance of these findings. Particular emphasis is placed on OCD, in view of the neuroanatomical impairment that this condition shares with HD, the symptomatic similarities between these disorders, and recent findings of an excess of OCD in HD-affected families. We hypothesize that precise characterization of the psychiatric status of some HD patients showing psychopathological manifestations and their families might help to distinguish different clinical subtypes of the disorder. This approach could hold promise in improving the management of HD in the future. PMID- 10689593 TI - It takes a village: caring for a traumatized art student. PMID- 10689594 TI - A model for the assessment of violence. PMID- 10689595 TI - Problem-based learning and psychiatry residency education. PMID- 10689596 TI - [Dexpanthenol for dry skin. Regeneration of damaged permeability barrier of the skin]. PMID- 10689597 TI - [Testosterone substitution in male senescence]. PMID- 10689598 TI - Knowledge development in health & social work. PMID- 10689599 TI - Culturally sensitive social work practice with Arab clients in mental health settings. AB - Several culturally specific practical considerations should inform social work interventions with ethnic Arab peoples in Arab countries or in Western nations. These include taking into account gender relations, individuals' places in their families and communities, patterns of mental health services use, and, for practice in Western nations, the client's level of acculturation. Such aspects provide the basis for specific guidelines in working with ethnic Arab mental health clients. These include an emphasis on short-term, directive treatment; communication patterns that are passive and informal; patients' understanding of external loci of control and their use of ethnospecific idioms of distress; and, where appropriate, the integration of modern and traditional healing systems. PMID- 10689600 TI - Risk factors associated with PTSD and major depression among Cambodian refugees in Utah. AB - The study reported in this article is a secondary analysis of data collected from a random sample of 124 Cambodian adults, ages 18 to 76 years. Participants were interviewed about their mental health status and factors associated with a diagnosis of posttraumatic stress disorder (PTSD) or major depression. From analysis of the data, the following risk factors were identified with PTSD and depression: experiencing a greater number of war traumas increased the risk of both PTSD and major depression; experiencing a greater number of resettlement stressors during the past year increased the risk of both PTSD and major depression; and having financial stress increased the risk of major depression. PMID- 10689601 TI - Depression among victims of south Mississippi's methyl parathion disaster. AB - Human-induced disasters have long been considered responsible for a wide array of physiological, psychological, and economic distress. This study examined depressive symptoms among victims of south Mississippi's methyl parathion disaster. Results indicated that irrespective of the level of methyl parathion contamination in respondents' dwellings, more than half the victims interviewed reported depressive symptoms at levels suggesting probable clinical depression. Those at greatest risk of depressive symptoms were people who had been exposed to the neurotoxin for the longest period of time, among whom there was an overrepresentation of women and African Americans. Despite high statistical levels of depression, few victims used mental health services. Implications for social work's response to human-induced disasters are provided. PMID- 10689602 TI - Adolescents with sickle cell disease: determinants of support group attendance and satisfaction. AB - Support groups have the potential to mitigate some of the developmental and condition-specific psychological and social issues common to sickle cell disease (SCD), yet little is known about how adolescents with SCD view and use these groups. As part of a larger study, 79 adolescents with SCD completed questionnaires assessing reasons for attending or not attending support groups, level and type of help received from group participation, group satisfaction, and attendance. This article reports on findings descriptively. Professionals can enhance their effectiveness as providers of social and mental health services by understanding the role that support groups play in the overall well-being of adolescents with SCD. PMID- 10689603 TI - A deconstructive turn in chronic pain treatment: a redefined role for social work. AB - Chronic pain treatment programs in North America are based predominantly on a behavioral science model of contingency management, whereby the focus of treatment is directed toward the psychological aspects of pain. Treatment objectives are designed to eliminate reinforcing environmental contingencies, thus changing pain behavior and reestablishing well behavior. The purpose of this article is to displace the contingency management model with deconstruction and thus present an alternative conceptualization based on the experiences of chronic pain sufferers. Social work's value base of self-determination and empowerment upholds these challenges to the predominant treatment model. Alternative social work interventions are explored. PMID- 10689604 TI - The power to choose: supports for families caring for individuals with developmental disabilities. AB - In an exploratory study of family support services in Massachusetts, three focus groups were convened to obtain the perspectives of parents caring for individuals with developmental disabilities and living at home. This article summarizes key themes that emerged from the group discussions: effects of family supports on family life, flexibility of supports, barriers, unmet needs, and recommendations for change. Social workers and health care professionals can enhance the well being of people with developmental disabilities and their families by addressing the needs of the entire family, facilitating family choice and control of supports, and helping families navigate the complex service system. PMID- 10689606 TI - Quality of life and mental health services. PMID- 10689605 TI - A "society for all ages": saving Social Security and Medicare. PMID- 10689607 TI - Subjective quality of life in female in-patients with depression: a longitudinal study. AB - This study investigated Subjective Quality of Life (SQOL) in 42 women with depression, 70 women with alcoholism, and 73 women with schizophrenia within 3 weeks after hospital admission. Twenty-eight of the depressive patients were re examined after 6 months. SQOL was assessed using the German version of the Lancashire Quality of Life Profile. On average, depressive women expressed dissatisfaction with life as a whole and with 4 out of 8 life domains, and had a lower SQOL than the other two diagnostic groups. Differences remain statistically significant when the influence of age and anxiety/depression is controlled for. SQOL in depressive women improved significantly within the follow up period. Positive SQOL change was moderately correlated with an improvement of depressive symptoms. The results indicate that depressive women after hospital admission express an unusually low SQOL, which seems to have some diagnostic specificity and improves over time. Changes in depressive symptoms do not fully explain SQOL changes. PMID- 10689608 TI - Predictors of subjective quality of life in schizophrenic patients living in the community. A Nordic multicentre study. AB - As part of a Nordic multi-centre study investigating the life and care situation of community samples of schizophrenic patients the aim of the present part of the study was to examine the relationship between global subjective quality of life and objective life conditions, clinical characteristics including psychopathology and number of needs for care, subjective factors such as satisfaction with different life domains, social network, and self-esteem. A sample of 418 persons with schizophrenia from 10 sites was used. The results of a final multiple regression analysis, explaining 52.3% of the variance, showed that five subjective factors were significantly associated with global subjective quality of life, together with one objective indicator, to have a close friend. No clinical characteristics were associated with global subjective quality of life. The largest part of the variance was explained by satisfaction with health, 36.3% of the variance, and self-esteem, 7.3% of the variance. It is concluded that the actual relationship between objective life conditions and subjectively experienced quality of life still remains unclear. Furthermore, it seems obvious that personality related factors such as self-esteem, mastery and sense of autonomy also play a role in the appraisal of subjective quality of life, which implies that factors like these are important to consider in clinical and social interventions for patients with schizophrenia in order to improve quality of life for these persons. PMID- 10689609 TI - Subjective quality of life, psychopathology, satisfaction with care and insight: an exploratory study. AB - We have investigated the determinants of global subjective quality of life (GSQOL) using data from a controlled trial of intensive case management for the severely mentally ill. In a multiple regression analysis depression, age, objective quality of life and domain-specific quality of life together explained 59% of the variance in GSQOL. GSQOL was not significantly correlated with measures of positive psychotic symptomatology, insight or attitudes to treatment but was correlated with satisfaction with care (r = .21, p = .011). Change in GSQOL over 18 months was correlated with change in domain-specific quality of life (r = -.45, p = .002) and depression (r = -.43, p = .01): multiple regression analysis confirmed that both variables had an independent effect on change in GSQOL. PMID- 10689610 TI - Quality of life and new antipsychotics in schizophrenia. Are patients better off? AB - The recent introduction of several antipsychotic medications has raised expectations for better pharmacological management of schizophrenia. Although conventional and new neuroleptics (Risperidone, Olanzapine, Seroquel and soon to be released Ziprasidone) are generally comparable in terms of efficacy; the new antipsychotic medications possess a better side-effects profile and are overall, much better tolerated. The reintroduction of Clozapine as an effective antipsychotic for treatment refractoriness has also improved management for a segment of the schizophrenic population who failed to respond adequately to other antipsychotic medications. Such increased benefits from new antipsychotic medications come with a higher acquisition cost that has somewhat strained the historically low psychiatric budgets. The question then was whether the expected benefits of the new antipsychotics can offset the high cost of these medications in the long-term. In that context, quality of life assessment has provided a tool for the comparative analysis of new and conventional antipsychotic medications, particularly regarding their impact on functional status and satisfaction. In a recently concluded study, we demonstrated that the new antipsychotic medications are subjectively much better tolerated and have a more favourable impact on quality of life compared with conventional neuroleptics. The ultimate question is whether such favourable benefits can translate in the future into better compliance with medications and improved long-term outcomes. PMID- 10689611 TI - Therapeutic relationships and quality of life: association of two subjective constructs in schizophrenia patients. AB - Subjective quality of life is an important criterion in outcome evaluation that has been well-researched in psychiatry. By comparison, the therapeutic relationship which may also be subjectively assessed has been relatively neglected as an outcome criterion although it has predictive power in relation to outcome. This exploratory study investigated subjective quality of life and therapeutic relationships in first-admission (N = 90) and long-term (N = 168) schizophrenia patients, each at two points of time. The follow-up period was 9 months for the first-admission sample and 1.5 years for the long-term sample. A significant relationship was found between global assessments of quality of life and therapeutic relationships in long-term, but not in first-admission patients. This finding was consistent at both assessments, suggesting that therapeutic relationships may become more central to quality of life in long-term care situations and that patients' views of this relationship are increasingly embedded in their overall appraisal of life. PMID- 10689612 TI - Measuring quality of life in secure care: comparison of mentally ill and personality disordered patients. AB - Improving quality of life for patients is emerging as a legitimate goal for UK inpatient forensic mental health services. The Lancashire Quality of Life Profile (LQOLP), which has been used widely to measure well-being in community settings, was trialed on an inpatient population being cared for under conditions of high security. Two groups of male patients, drawn from within the same institution but with markedly different clinical conditions, i.e. schizophrenia (N = 47) and personality disorder (N = 48), were interviewed using the LQOLP. Although both groups had been cared for under largely similar environmental conditions over similar lengths of time (9.5 years), the subjective global well-being of the two groups differed systematically as did other objective and subjective well-being measures. However, analysis found that the variations in global well-being could not be attributed readily to factors covered by the interview, including either current mood or personality. Possible reasons for these findings and implications for the use of the LQOLP under conditions of high security are discussed. PMID- 10689613 TI - The impact of the interviewer-interviewee relationship on subjective quality of life ratings in schizophrenia patients. AB - Subjective quality of life (SQOL) ratings are usually based on interviews. This study examined in which way patients' ratings differ depending on whom they are interviewed by. SQOL was assessed in 78 schizophrenia patients in an out patient clinic and in sheltered living arrangements. Using patients randomly allocated to two interview situations: one group was interviewed by external researchers, the other group by their case managers. On average, more favourable ratings were elicited by case managers. Some of the differences were statistically significant and substantial in size. Yet, opposing differences were also found regarding some life domains in one group. It may be concluded that a significant impact of the interviewer-interviewee relationship on SQOL ratings may exist, but that it is not consistent, unidirectional and uniform regarding life domains and across different settings and samples. PMID- 10689614 TI - Rehabilitation programmes and quality of life in severe mental illness. AB - Quality of life is increasingly identified as a key outcome measure for evaluating the efficacy of community mental health services and novel antipsychotics. However, there is a relative paucity of research on the impact of rehabilitation programmes on quality of life. This report outlines the results of two 'naturalistic' studies carried out in a catchment area psychiatric service to evaluate the benefits associated with a supported employment programme and a psychosocial/educational intervention. The findings suggest that outpatient based programmes which provide opportunities for vocational or prevocational rehabilitation may have significant quality of life benefits for individuals with severe mental illness. PMID- 10689615 TI - An evaluation of the impact of clubhouse membership on quality of life and treatment utilization. AB - A group of clubhouse users matched with similar patients (not clubhouse users) in a neighbouring area were compared in terms of quality of life (Lancashire Quality of Life Profile), service utilization and treatment costs over a two year period. The clubhouse group achieved a reasonable employment status and good social relationships, and advantages in subjective well-being favoured the clubhouse group. Over two years the pattern of service utilization and costs also favoured the clubhouse group. When the two groups were disaggregated for employment status the group with least treatment utilization and lowest costs was the employed clubhouse group. PMID- 10689617 TI - [Corticosteroid hormones: mechanisms involved in the recognition of aldosterone by mineralocorticoid receptors]. AB - Aldosterone and cortisol, the major mineralocorticoid and glucocorticoid hormones in humans, are structurally very closed. Both hormones bind to the mineralocorticoid receptor (MR) with the same affinity. Nevertheless MR is preferentially activated by aldosterone, suggesting that the binding of these two hormones to MR involved some distinct contacts. We constructed a tridimensional model of the ligand-binding domain of the human MR, by taking as a template the structural data of the retinoid receptor associated with its ligand. The MR model allowed the identification of several residues involved in the interaction with aldosterone and cortisol. The residues Gln 776 and Arg 817 make hydrogen bonds with the 3-keto function and the residue Asn 770 with the C21-hydroxyl group. Analyses of the wild type and mutant MRs activities in response to corticosteroids bearing hydroxyl groups at various steroid skeleton position led to the following conclusions: 1) the interaction between the residue Asn 770 and the C21-hydroxyl group of corticosteroids is determinant for stabilizing the active MR conformation and 2) the stability of this conformation is enhanced by the 11-18 hemiketal group of aldosterone whereas it is decreased by the 11 beta- and 17 alpha-hydroxyl groups of cortisol. These results are discussed in the light of a model for the MR activation process. PMID- 10689616 TI - [The head inducer Cerberus in a multivalent extracellular inhibitor]. AB - Cerberus encodes for a secreted protein which when overexpressed ventrally in a Xenopus embryo induces head differentiation without trunk (Bouwmeester et al., 1996). We have recently shown that Cerberus can bind BMP-4 (Bone Morphogenetic Protein-4), Xnr-1 (Xenopus Nodal-related 1) and Xwnt-8 in the extracellular space (Piccolo et al., 1999). We present here studies showing that Cerberus does not have a receptor nor a dedicated transduction pathway but rather acts as an extracellular inhibitor. Our results suggest that the action of Cerberus in head induction can be explained by an inhibitory activity upstream of the Nodal related and BMP-4 receptors. In addition, using dominant negative receptor mutants which block both the Xnr-1 and BMP-4 transduction pathways, we show that this double inhibition is sufficient for head induction in ventral mesoderm explants. PMID- 10689618 TI - [Functional interaction of HSP90 with steroid receptors]. AB - Hsp90 (Heat Shock Protein 90) is a component of the inactive and metastable hetero-oligomeric structure of steroid receptors. Recent data on Hsp90 structure and function as a stress protein and dedicated molecular chaperone are here reviewed with a particular focus on Hsp90 chaperone cycle interfering with steroid receptor action. The dual role of Hsp90 as a positive and negative modulator of steroid receptor function is considered along the activation desactivation process of the receptors. It is proposed that Hsp90 chaperone machinery assists the receptor during its synthesis thus avoiding collapse and facilitating an open structure able to bind ligand efficiently. Moreover, it is suggested that Hsp90 may help the folding of the hydrophobic core of the receptor around the ligand and finally Hsp90 may chaperone the receptor after the dissociation of the ligand. PMID- 10689619 TI - [Interactions between the glucocorticoid receptor and transcription factors]. AB - Transcriptional regulation by glucocorticoids is mediated through an intracellular glucocorticoid receptor which transmits hormone signal to the nucleus. Two types of mechanisms have been attributed for the hormonal regulation of gene promoters. The first one requires DNA binding of the activated receptor to a specific element called GRE (Glucocorticoid Response Element) found in the promoter regions of target genes and the second one involves a direct cross-talk of the GR with transcription factors. Both mechanisms are dependent on the promoter configurations, their chromatin structure or the components of the transcriptional complexes involved in the interaction with the GR. These distinct features specify the activity of the GR as a repressor or activator of transcription. PMID- 10689620 TI - [Glucocorticoids and acute phase proteins]. AB - Glucocorticoids as well as acute phase proteins participate in non-specific host defence as well as in restoring host integrity after injury. Plasma levels of both compounds augment during the inflammatory reaction. However, glucocorticoids also have physiological effects that share similar molecular mechanisms with the family of steroids. During the inflammatory reaction, and for participating in host defense, glucocorticoids, together with augmented cytokines, use new signalling pathways. In doing so, they participate in the positive or negative control of inflammatory mediator synthesis. For example, they induce the synthesis of acute phase proteins in synergy with interleukin 6, interleukin 1 and TNF alpha. PMID- 10689621 TI - [Interactions between glucocorticoids and anti-inflammatory peptides]. AB - Both pro- and anti-inflammatory mediators regulate the anti-inflammatory actions of glucocorticoids, in part by modifying the binding of glucocorticoids to specific receptors. For instance, somatostatin has been shown to increase glucocorticoid binding and signaling in macrophages. The mechanism of this regulation does not require an increased expression of glucocorticoid receptors but, rather, a stabilization of glucocorticoid receptor-associated heat shock protein 90. This is related to a decrease in calpain activity. Thus calpain inhibition may offer a new and exciting possibility for enhancing the anti inflammatory efficiency of glucocorticoids. PMID- 10689622 TI - [Temporal regulation of gene expression]. AB - Molecular biology gives a static--not a dynamic--vision of the mechanisms regulating gene expression. Genetics already gave to time a limited place in the explanation of living phenomena. Such a static vision is supported by the techniques--such as X-ray crystallography--used by the biologists. However time is an important parameter in the control of gene expression during the cellular response to external signals, during life and aging of organisms or even in the succession of living forms which takes place in evolution. Models are slowly moving, due to the eruption of new technologies giving access to the fast events which occur inside living cells. A new dynamic vision is progressively replacing the old one. The consequences of these changes on the form of the future biology remain still unknown. PMID- 10689623 TI - [Management of time by the cell and by the organism]. AB - Time-dependent regulations of cells and organisms can be analysed at different levels. One of these levels is the periodicity of cell functions such as cell division, metabolic processes (generation of ATP by glycolysis or oxidative mitochondrial processes) and the biosynthesis of cell constituents. Studies carried out on unicellular eukaryotes revealed the periodic, oscillatory nature of most of these processes. Time constants of these reactions vary from nanoseconds to hours-days, necessitating coupling mechanisms. Comparative studies revealed the coupling of the rapid processes (mitochondrial ATP generation) to the slower rhythms of the biosynthetic processes of macromolecules. Adenine nucleotides are involved in the coupling mechanisms between rapid and slow processes ("the slow dance of life to the music of time"). The mechanisms underlying these rhythmic processes involve either key allosteric regulatory enzymes (PFK for glycolysis) or "desensitization" of receptors by phosphorylation dephosphorylation. At the organismic level the study of rhythmic processes is illustrated by the periodicity of heart beats, shown to exhibit multifractality, following apparently the formalism of deterministic chaos. Another example is the rhythmic oscillatory discharges of neuronal networks. The existence of subrhythmes mostly of epigenetic nature, facilitated probably the progressive adjustment of cells during evolution to the slow increase of day time since the separation of the moon from the earth. We analysed the mechanisms underlying the decline of these processes during aging. Loss of receptors or/and their uncoupling from their transmission pathway appear to be involved in most of these processes of decline. One conclusion of this review is the importance of epigenetic mechanisms both in the genesis and in the decline of these rythmic processes involved in time keeping by the cell. PMID- 10689624 TI - [Time of consciousness, consciousness of time]. AB - Time shapes our behavior: we must estimate the duration of events in our environment in order to anticipate changes and time our activity in function of these changes. However, there is no sensory modality devoted to the perception of time, therefore the question is to know which mechanisms underlie the consciousness that time flows and allow us to estimate time precisely. This article proposes a brief overview of psychological, neuropsychological and brain imaging studies which rely on theoretical models postulating the existence of an internal timer. These studies examine the different components--time base, counter and memory--of this timer: particularly they are aimed at characterising the relationships between the evolution of these components with age or their pathological alterations and changes in temporal judgements. They also attempt to specify the neural bases of these components. From this brief overview comes the idea that, if an internal timer exits, it does not mark objective time but a multitude of subjective times. PMID- 10689625 TI - [Regulation of cell activity by the extracellular matrix: the concept of matrikines]. AB - The activity of connective tissue cells is modulated by a number of factors present in their environment. In addition to the soluble factors such as hormones, cytokines or growth factors, cells also receive signals from the surrounding extracellular matrix (ECM) macromolecules. Moreover, they may degrade the ECM proteins and liberate peptides which may by themselves constitute new signals for the surrounding cells. Therefore, an actual regulation loop exists in connective tissue, constituted by peptides generated by ECM degradation and connective tissue cells. The term of "matrikine" has been proposed to designate such ECM-derived peptides able to regulate cell activity. In this review, we summarize some data obtained in our laboratory with two different matrikines: the tripeptide glycyl-histidyl-lysine (GHK) and the heptapeptide cysteinyl asparaginyl-tyrosyl-tyrosyl-seryl-asparaginyl-serine (CNYYSNS). GHK is a potent activator of ECM synthesis and remodeling, whereas CNYYSNS is able to inhibit polymorphonuclear leukocytes activation and decrease the invasive capacities of cancer cells. PMID- 10689626 TI - [Role of lipid second messengers involved in the response of leukemic cells to anticancer drugs]. AB - Simple clinical observation suggests that while anti-leukemia agents are efficient at eradicating blasts cells in terminal division, as illustrated, in the case of acute myeloid leukemia, by the high complete remission rate (70%); these agents are relatively inept at eliminating leukemic myeloid progenitors as suggested by the high level of recurrence. This interpretation underlines the apparently natural chemoresistance of cells which compose the myeloid leukemia progenitor compartment. Over the past few years, several studies have shown that similar cellular damage can lead to divers effects such as rapid apoptotic death, differed mitotic death, or a transitory cytostatic effect. Cell response to damage is regulated by a complex and highly regulated network of intracellular signals including cell death signals mediated by ceramide and cell survival signals mediated (at least in part) by diacylglycerol and phosphoinositide-3 phosphates. Cellular fate relies on the balance between these two signaling pathways. This hypothesis opens several prospects on pharmacological manipulation aimed at either favoring cell death or at conferring resistance to anti-cancer agents. PMID- 10689627 TI - Otilonium bromide: a selective spasmolytic for the gastrointestinal tract. AB - Experimental studies have shown that otilonium bromide (OB) inhibits both baseline and chemically or physically stimulated gastrointestinal motility. The spasmolytic activity of OB in the gastrointestinal tract occurs at doses that do not affect gastric secretion or produce typical atropine-like side-effects. The mechanism of action is composite: interference with calcium ion movement from intra- and extracellular sites; blockade of calcium channels; and binding to muscarinic receptors and tachykinin neurokinin-2 receptors. Pharmacokinetic studies have shown that OB accumulates in the lower intestine and has poor systemic absorption. Clinical studies have confirmed OB as a potent spasmolytic drug with a good tolerability profile. Studies in patients with irritable bowel syndrome demonstrated OB to be superior to placebo and reference drugs in parameters such as pain, abdominal distension and motility. The composite and local mechanism of OB action reduces hypermotility and modulates visceral sensation: factors thought to be responsible for pain improvement recorded in clinical trials. The compound is marketed worldwide and no serious adverse events have been reported as yet, confirming its excellent tolerability. PMID- 10689628 TI - Efficacy and safety of trazodone versus clorazepate in the treatment of HIV positive subjects with adjustment disorders: a pilot study. AB - The efficacy of trazodone and clorazepate to relieve anxiety and depressive symptoms in 21 HIV-positive subjects with adjustment disorders was determined in a 28-day single-centre, randomized, double-blind study. Subjects were evaluated using the Hospital Anxiety and Depression Scale, the Revised Symptom Checklist, the European Organization for Research and the Treatment of Cancer Quality of Life Questionnaire, and a binary criterion based on the Clinical Global Impression. The incidence of successful treatment was 80% for trazodone compared with 64% for clorazepate; the sample number was too small to establish a significant difference. Bayesian analysis revealed the probability of making a wrong decision in prescribing trazodone rather than clorazepate reduced from 35% to 18% in this small sample. Clinical evaluations using the different scales suggest some benefit from trazodone, although this was not significant. Safety of both treatments was similar. Trazodone is devoid of the risk of abuse and dependence, and may be a valuable alternative to benzodiazepines for the treatment of HIV-related adjustment disorders. PMID- 10689629 TI - The effect of cross-linked hyaluronate hydrogel on the reduction of post-surgical adhesion reformation in rabbits. AB - The effects of cross-linked hyaluronate hydrogel and liquid sodium hyaluronate on post-surgical adhesion reformation were examined using a rabbit model. Primary adhesions in the ileocaecal region of Japanese white rabbits were induced by mechanical and chemical irritants during laparotomy. After 1 month the primary adhesions were lysed by microsurgery and cross-linked hyaluronate hydrogel or liquid sodium hyaluronate was applied to the lysed lesions. After 10-14 days the area of adhesion reformation was measured to assess any inhibitory effect of the test materials. Rabbits treated with cross-linked hyaluronate hydrogel showed a significant reduction in adhesion reformation area compared with liquid sodium hyaluronate or physiological saline treatment, and the area reduced to (mean +/- standard deviation) 0.6 +/- 1.95% of the original lesion. In a separate study, histological evaluation of rabbits treated with cross-linked hyaluronate hydrogel revealed a better healing pattern and a lower inflammatory response compared with controls. All these findings suggest cross-linked hyaluronate hydrogel may be a valuable anti-adhesion material to prevent post-surgical adhesion in abdominal or pelvic surgery. PMID- 10689630 TI - Use of fibrin adhesive to reduce post-surgical adhesion reformation in rabbits. AB - Following surgery on fallopian tubes, the development of adhesions is a natural consequence of wound healing and may result in infertility. Using a rabbit model, we evaluated the anti-adhesive properties of a sponge-like equine collagen sheet (TachoComb), which is coated on one side with human fibrinogen and bovine thrombin. TachoComb is applied by affixing the sheet over the area of perforation or bleeding and acts as a haemostatic agent, capable of sealing perforations to prevent leakage. In our rabbit model, adhesions were induced by mechanical and chemical irritants during laparotomy. After a 1-month recovery period, adhesions were lysed using microsurgical techniques and TachoComb, or physiological saline applied. Evaluation of adhesion reformation was determined after a minimum of 10 days. TachoComb significantly reduced the area of adhesion reformation compared with rabbits treated using physiological saline only. Our study demonstrated that TachoComb is effective not only as a haemostatic agent, but is also capable of reducing adhesion reformation. PMID- 10689631 TI - Clinical evaluation of a haemostatic and anti-adhesion preparation used to prevent post-surgical adhesion. AB - TachoComb consists of equine collagen in a sponge-like form coated on one side with human fibrinogen and bovine thrombin. This product functions as a haemostatic and physical barrier to inhibit post-surgical adhesion. In this study, we investigated TachoComb to control oozing in 16 patients who required haemostasis. Evaluation of post-surgical adhesion by second-look laparoscopy was performed at 3 months and 7 months after initial surgery. Observation via laparotomy during Caesarean section was also performed at 13 months, 3 years and 4 years after initial surgery. In all but one patient, no macroscopic evidence of TachoComb persistence was found. Furthermore, no de novo adhesions were detected at the TachoComb application site. We have thus demonstrated that TachoComb can be used to control oozing haemorrhage effectively from surgical sites and can prevent adhesion formation at the application site, and may thus be an effective method of preventing adhesion-induced infertility. PMID- 10689632 TI - Use of androgens and oestrogens in adolescents--a review of hormone replacement treatment. PMID- 10689633 TI - Endocrine activity during sleep. AB - Almost all functions of humans are subject to cyclic changes and are governed by the nervous system. Most rhythms are driven by an internal biological clock located in the hypothalamic suprachiasmatic nucleus (SCN) and can be synchronized by external signals such as light-dark cycles. Homeostatic activities such as body temperature, blood volume, water balance and sleep, are rhythmic. Likewise, most hormones are secreted in a rhythmic fashion. Both sleep and circadian effects interact to produce the overall rhythmic pattern of the pituitary and pituitary-dependent hormones. Some of the 24-h hormonal rhythms depend on the circadian clock (ACTH, cortisol and melatonin), or are sleep related (prolactin and TSH). GH secretion is influenced by the first slow wave sleep (SWS) episode at the beginning of the night. Pulses of prolactin and GH are positively linked to increases in delta wave activity, i.e. deepest phases of sleep, occurring primarily during the first third of the night. Pulses of TSH and cortisol are related to superficial phases of sleep. As a result of the consolidation of the sleep period in humans, the wake-sleep transition is associated with physiological changes with the endocrine system being part of the adaptive mechanism to reduce physical activity during sleep. PMID- 10689634 TI - Growth hormone deficiency type IB caused by cryptic splicing of the GH-1 gene. AB - We have found a novel mutation in intron 4 of the GH-1 gene in a Bedouin kindred with isolated growth hormone deficiency type IB (IGHD IB). RFLP analysis suggested linkage between the GH-1 gene and IGHD. Nested PCR amplification followed by single stranded conformation polymorphism (SSCP) analysis indicated sequence variation between introns 2 and 4. Sequencing showed a G-->C transversion at the fifth base in the splice donor region of intron 4. Affected individuals were homozygous for the mutation, which creates a new Mae III restriction site. Reverse transcription and PCR of GH-1 transcripts in EBV transformed lymphocytes indicated predominance of a species lacking 73 bp of exon 4. Amplification with a bridging primer showed that the same mRNA species is present in lymphocytes from normal individuals. The first 102 amino acids of the predicted protein are identical to wild-type GH, but the next 94 amino acids are completely divergent. PMID- 10689635 TI - An atypical kindred with X-linked adrenal hypoplasia congenita, normal puberty, and normal Dax-1 promoter and coding sequence. AB - We report a Chinese kindred with an atypical sex-linked form of isolated adrenal hypoplasia without hypogonadotropic hypogonadism. Evidence of sex linkage was supported by DNA analysis using three polymorphic markers from the X-chromosome: a restriction fragment length polymorphism 200 kb centromeric of the DAX-1 gene, a tetranucleotide repeat marker in the DAX-1 promoter (DAX-P), and a microsatellite in the Duchenne muscular dystrophy locus (3'-19). This pedigree therefore presents the novel phenotype of sex-linked hypoadrenalism without hypogonadotropic hypogonadism, with evidence of possible linkage to the DAX-1 gene. However, all three affected individuals were examined for mutations in the DAX-1 gene, and found to have no sequence anomalies in the coding region, splice sites or 5' non-coding region. This presentation may be due to a defect in the DAX-1 gene outside its known coding region, possibly modulated by functional polymorphisms at other loci, and/or environmental effects, or to a defect in a novel gene on the X chromosome which selectively influences adrenal development. PMID- 10689636 TI - Persistent hyperinsulinemic hypoglycemia of infancy: long-term outcome following subtotal pancreatectomy. AB - BACKGROUND: Persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is the most common cause of persistent hypoglycemia in infants. The current standard treatment is subtotal pancreatectomy (Px). However, the long-term outcome following surgery needs further attention. METHODS: We analyzed 10 children (7 M, 3 F) with PHHI who underwent partial (65-80%) and subtotal (81-95%) Px. Follow-up ranged from 2 to 9.4 yr (mean = 4.2 yr). We divided them into 2 groups based upon the age at onset of hypoglycemia: early (< 1 mo) and late (> or = 1 mo). RESULTS: The seven patients in the early-onset group underwent 85-95% Px between ages of 18 d and 3 mo. Three of them initially treated by 85-90% Px had persistent hypoglycemia postoperatively. Two out of three required a 2nd operation with 95% Px for controlling hypoglycemia, though both still had persistent hypoglycemia and required medication to control blood glucose. The remaining four had 95% Px and had maintained euglycemia postoperatively. One patient developed diabetes 6 yr after surgery. Six of seven patients had delayed development and subnormal IQ. Three patients of the late-onset group (3 mo, 6 mo and 4 yr) underwent partial Px (80%, 65% and 65%, respectively) and maintained euglycemia postoperatively. Despite 65% Px, one developed diabetes 3 yr after surgery. CONCLUSIONS: These results suggest that children with early-onset hypoglycemia have more severe hyperinsulinism than those with late-onset hypoglycemia. The former require 95% Px for maintaining euglycemia, but long-term complications with diabetes may be common. In contrast, the latter require lower percentage Px which may reduce the incidence of diabetes in the future. PMID- 10689637 TI - Length velocity acceleration at 9 months of age in a representative birth cohort of Dutch infants. AB - According to the ICP (infancy-childhood-puberty) growth model, statural growth can be divided into three partially superimposed components assumed to represent different physiologic mechanisms. This model predicts a sudden acceleration of length velocity (LV) at the onset of the childhood component around 9 months. The existence of such an infancy-childhood growth spurt has not yet been firmly corroborated by epidemiological studies. In the present study length measurements were made at the target ages of 1, 3, 6, 9, 12, 15, 18 and 24 months in a birth cohort of 2034 infants. In order to check whether length growth showed a continuous smooth pattern, different mathematical models were fitted to the individual growth curves. The models included Count and Guo functions, 5th order polynomial and combinations of 5th order polynomial with the logarithmic term of the Count function and the square root term of the Guo function. We showed that in boys and girls there is a small but systematic lack of fit of the mathematical modeling, due to a sudden acceleration of LV around 9 months. In addition there was an increase in variation of attained length at this age. Comparison of unbalanced ANOVA models with and without addition of dummy variables for the target ages confirmed that there was an acceleration around 9 months that, if corrected for, leads to a significantly improved model fit (likelihood ratio test p < 0.0001). In absolute terms of LV, the misfit at 9 months was not greater than 0.5 cm/year on average. We conclude that the results of this study support the existence of a late infancy growth spurt. In our opinion, however, the magnitude of the phenomenon does not legitimate construction and use of discontinuous growth references such as the ICP reference. PMID- 10689638 TI - Penile length of newborns in Singapore. AB - Micropenis is an important sign in congenital hypopituitarism and various disorders. Documented norms for penile length exist only for babies of Caucasian and Middle-Eastern origin. This study was carried out to establish such norms for Asian newborns. We studied 228 male live births within their first three days of life. Stretched penile lengths were marked off on unmarked wooden spatulas, which were placed vertically along the dorsal aspect of the penis, with one rounded end on the pubic bone. The mean penile length +/- S.D. for the full-term Asian baby was 3.6 +/- 0.4 cm. Race had a significant effect: Chinese 3.5 cm, Malay 3.6 cm and Indian 3.8 cm. Penile length correlated with birth weight and gestational age. Asian babies thus have similar norms to Caucasian babies. An Asian newborn whose penis measures less than 2.6 cm has micropenis and may need prompt investigation for underlying endocrine disorders. PMID- 10689639 TI - Longitudinal assessment of L-T4 therapy for congenital hypothyroidism: differences between athyreosis vs ectopia and delayed vs normal bone age. AB - To study the effects of LT4 dose on thyroid hormone serum levels, a prospective, longitudinal and comparative study was designed, including 56 term eutrophic 1-89 day-old infants with congenital hypothyroidism (CH) detected by neonatal screening. Patients were divided into four groups according to delayed or normal bone age at birth, and athyreosis or ectopic thyroid. All received an initial dose of 50 micrograms/day (12.9-13.7 micrograms/kg/day) LT4 and were followed bimonthly (first year) and quarterly (second year) with thyroid profile and bone age determinations at 6, 12 and 24 months. At diagnosis, hormone levels were higher in cases of ectopia than in athyreosis (p < 0.001), and T4 was lower in children with delayed than in normal bone age at birth (p < 0.05). During treatment, all groups were clinically euthyroid despite T4 and FT4 serum levels higher than the upper normal limit (p < 0.0.001), though T3 and FT3 were within the normal limit (p > 0.05). TSH normalized within 8 weeks. Bone age accelerated at 2 years in eight children of the bone age delayed group. No patient had craniosynostosis. PMID- 10689640 TI - Cord blood leptin levels: relationship to body weight, body mass index, sex and insulin and cortisol levels of maternal-newborn pairs at delivery. AB - To investigate leptin and to which factors it is related during the perinatal period, we measured serum leptin levels of 46 mothers at delivery, umbilical cord blood and infants on the third day of life. Maternal leptin was higher than in cord (p < 0.001), and did not correlate with maternal age, body weight, body mass index, weight gain during pregnancy, serum glucose, cholesterol, triglycerides, CPE, cortisol or HbA1c levels, nor any biochemical values or anthropometric data of the newborns (p > 0.05). In cord blood, leptin was significantly higher than in 3 day-old infants (p < 0.05), and correlated only with maternal insulin and glucose (r = 0.5, p < 0.01 and r = 0.4, p < 0.05, respectively). In 3 day-old infants, leptin did not correlate with any clinical data (p > 0.05). Leptin was not different in the two sexes (p > 0.05). Serum leptin levels were not related to adiposity of the mother-infant pairs or neonatal growth, and were not different in the two sexes during the perinatal period. PMID- 10689641 TI - Intensive diabetes management in adolescents with type 1 diabetes: the importance of intensive follow-up. AB - Minimal information exists on the education and follow-up required to successfully initiate intensive diabetes management (IDM) in adolescents with type 1 diabetes. We performed a retrospective analysis of HbA1c 3 and 15 months after initiation of IDM in two cohorts: (1) 17 patients who received individualised education in IDM and intensive early follow-up, and (2) 11 patients who participated in group education for initiation of IDM with standard follow-up. Entry HbA1c was higher in the individualised education patients (9.5 +/- 0.3% [mean +/- SE] versus 8.2 +/- 0.4%, p = 0.02). After 3 months of IDM, HbA1c improved in both cohorts reaching similar levels (individualised: 7.0 +/- 0.1%, p < 0.0001 vs entry; group: 7.3 +/- 0.2%, p = 0.05). During the following year, with routine follow-up for both cohorts, HbA1c levels rose approximately 1% as patients reverted to a multiple daily injection regimen. Irrespective of the educational approach, we believe maintenance of IDM and optimal HbA1c requires long-term intensive follow-up. PMID- 10689642 TI - Circulating L-selectin concentrations in children with recent-onset IDDM. AB - To clarify conflicting claims of altered serum concentrations of soluble L selectin (sCD62L) in recent-onset IDDM, sCD62L was measured in 89 children and adolescents with IDDM (35 recent-onset, 12 during the first year of insulin treatment, and 42 with long-standing (> 1 yr) treatment) alongside 124 controls. Children < 14 yr of age both with and without IDDM (n = 160) had grossly elevated sCD62L concentrations (20.2 +/- 4.9 nmol/l), as compared with adolescents (14-18 yr, n = 23; 15.9 +/- 3.9 nmol/l) and adults (> 18 yr, n = 30; 11.2 +/- 2.3 nmol/l) (p < 0.0001). Multivariate analysis confirmed the strong inverse association between age and sCD62L (p < 0.001) while revealing that sCD62L concentrations were slightly elevated in recent-onset IDDM, as compared with insulin-treated IDDM patients or nondiabetic controls (p = 0.028). Actual sCD62L concentrations in the 35 recent-onset IDDM patients were 22.2 +/- 4.9 nmol/L vs 19.6 +/- 3.6 nmol/l in 35 non-diabetic controls matched for age (p = 0.022). While this significant but small rise of systemic sCD62L reflects leukocyte activation, it is obscured by the inverse association between sCD62L and chronological age in children and adolescents. Therefore, determining sCD62L serum concentrations appears to be of limited value for clinical investigators caring for children and adolescents with IDDM. PMID- 10689643 TI - Analysis of cytokine mRNA expression in pancreatic islets of nonobese diabetic mice. AB - Nonobese diabetic mice develop type 1 diabetes in an age-related and gender dependent manner. Th1 (IFN-gamma and TNF-beta) and Th2 (IL-4 and IL-10) cytokine mRNA expression was analyzed in pancreatic islets isolated from female NOD mice with a high incidence of diabetes and male NOD mice with a low incidence of diabetes. The levels were measured at 5 time points from the onset of insulitis until the development of overt diabetes, using a semiquantitative reverse transcriptase PCR (RT-PCR) assay. IFN-gamma mRNA levels were significantly higher in the islets obtained from females than those of males, from 10 weeks of age. TNF-beta mRNA was expressed in both females and males between 5 and 15 weeks of age. However, TNF-beta mRNA levels were decreased in males at 20 weeks of age. In contrast, IL-4 mRNA levels were lower in females than in males. These results suggest that islet beta-cell destruction and diabetes in female NOD mice correlates with IFN-gamma and TNF-beta production in the islets, and that male NOD mice may be protected from autoimmune beta-cell destruction by down regulation of these cytokines. Furthermore, our findings also suggest that insulitis and beta-cell destruction are independently regulated: TNF-beta is more important in forming and maintaining the insulitis, while IFN-gamma has a more important role in beta-cell destruction. PMID- 10689644 TI - Thyroid agenesis associated with phalangeal anomaly. PMID- 10689645 TI - Serum inhibin B concentration in a prepubertal boy with gynecomastia and Peutz Jeghers syndrome. AB - Gynecomastia in boys with Peutz-Jeghers syndrome and Sertoli cell tumors of gonadal origin results from increased estrogen production due to increased aromatase activity within the testicular tumor. We present a prepubertal boy with Peutz-Jeghers syndrome, gynecomastia and bilateral neoplastic Sertoli cell proliferation in whom the only abnormal hormonal profile was increased concentration of inhibin B and Pro-alpha C in serum. PMID- 10689646 TI - Nonclassic 11 beta-hydroxylase deficiency: report of two patients and review. AB - Congenital adrenal hyperplasia (CAH) is well recognized as a disorder which can result in virilization of females, accelerated skeletal maturation and resultant adult short stature in both genders, and, in certain varieties, life-threatening adrenal crisis. Among the enzymatic defects resulting in CAH, nonclassic or partial 11 beta-hydroxylase deficiency is a relatively uncommon etiology. However, the subtlety with which it can present and the difficulties associated with its diagnosis can delay its identification and result in a significant reduction in adult stature. This paper describes the presentation and evaluation of two children with partial 11 beta-hydroxylase deficiency, discusses its pathogenesis, and compares the disorder with the more common varieties of congenital adrenal hyperplasia. PMID- 10689647 TI - Hand test AGG and AOS variables: relation with teacher rating of aggressiveness. AB - The purpose of this study was to determine the extent to which the Hand Test (Wagner, 1983) variables Aggression (AGG) and the Acting Out Score (AOS) were able to differentiate a group of children who were identified as aggressive and referred for psychological assessment by their teachers from a nonreferred, control group. Hand Test scores of 37 children who had consecutive referrals for psychological assessment because of aggressiveness were compared to the Hand Test scores of 37 children, matched on age and sex, from a nonreferred group. Through the use of an analysis of variance, AOS and AGG were found to significantly differentiate between the two groups. Spearman (rho) correlations between AGG and AOS scores with aggressive-referred status were rho = .45, p = .0001, and rho = .32, p = .006, respectively. Also, diagnostic efficiency statistics demonstrated moderate to high overall correct classification rates for AOS > or = 0 and AGG > or = 2 in identifying children in the aggressive-referred group. The results of this study provide support for the validity of the AGG and AOS scores in the assessment of aggressive behavior in children and demonstrate the utility of the Hand Test to identify aggressive tendencies in children. PMID- 10689648 TI - Relation between alexithymia and the five-factor model of personality: a facet level analysis. AB - The relation between alexithymia and both the domain and the facet level of the five-factor model (FFM) of personality was examined in a sample of 101 university students by using the Twenty-Item Toronto Alexithymia Scale (TAS-20; Bagby, Taylor, & Parker, 1994) and the Revised NEO Personality Inventory (Costa & McCrae, 1992c). Consistent with the alexithymia construct, the TAS-20 was positively correlated with Neuroticism (N) and negatively correlated with Extraversion (E) and Openness (O), whereas no significant relations were found with Agreeableness (A) and Conscientiousness (C). Analysis of the lower order traits (i.e., facets) of the FFM revealed that depression for N; positive emotions and assertiveness for E; feelings and actions for O; altruism, tender mindedness, and modesty for A; and competence for C predicted alexithymia. These results support the uniqueness of the alexithymia construct, which is represented by a cluster of traits across the dimensions and facets of the FFM. PMID- 10689649 TI - Development and initial validation of a brief mental health outcome measure. AB - Using a combination of classical test theory and Rasch item analysis, we developed a short scale designed to measure the effectiveness of mental health treatment across a wide range of mental health services and populations. Item development for the scale was guided by literature review and interviews with senior clinicians and with patients. Using 3 different samples consisting of inpatients, outpatients, and nonpatients, we reduced our initial item pool from 81 to 10 items. The 10-item scale had an alpha of .96 and showed strong correlations with commonly used measures of psychological well-being and distress. Our results suggest that the scale appears to measure a broad domain of psychological health. The scale appeared to lack ceiling and floor effects, and it discriminated between inpatients, outpatients, and nonpatients, suggesting the scale has excellent potential to be broadly responsive to a variety of treatment effects. In addition, the new scale proved to be sensitive to treatment changes in a sample of 20 psychiatric inpatients. Overall, the initial data suggest that we have developed a brief, sensitive outcome measure designed to have wide application across psychiatric and psychological treatments and populations. PMID- 10689650 TI - Normative and psychometric data on the body image assessment--revised. AB - After falling into disfavor in the early 1990s, the construct of body image, as measured by body-size estimation (BSE) techniques, has been the focus of increasing interest in the eating disorder literature because of recent theoretical, empirical, and methodological advances. However, no published BSE measure to date has been shown to be psychometrically sound, well normed, inexpensive, and straightforward. This article provides normative and psychometric data for an adapted silhouette BSE measure. Comprehensive normative data are presented on college women's cognitively and affectively based body-size estimates, as well as their desired body size and related discrepancy indexes (cognitive vs. desired, affective vs. desired, affective vs. cognitive). Preliminary data indicate that indexes from the new measure are moderately reliable over time, consistent with their theoretical link to fluctuations in body-related attitudes. Data also support the convergent validity of the measures. Affectively based BSE, alone or as part of a discrepancy measure with desired body size, was most strongly related to measures of eating pathology, body focus, body dissatisfaction, and depressed affect. PMID- 10689651 TI - Efficacy of the three Randomness Validity Scales for the Jesness Inventory. AB - Efficacies of the three randomness validity scales for the Jesness Inventory were investigated: the Jesness Variable Response Inconsistency scale (J-VRIN), the Variable Response scale (VR), and the Randomness scale (RD). Effectiveness was assessed by comparing the protocols of 93 male and 45 female delinquents ages 14 to 18 years screened for probable randomness, with a matched-pair MMPI-Adolescent with 500 computer-generated all- and half-random protocols. With the all-random set, for specificities of .90 or higher, scales showed sensitivities as high as .95 (VR), .90 (J-VRIN), and .14 (RD). With the half-random protocol, set sensitivities were .74 (VR), .70 (J-VRIN), and .07 (RD). Predictive power and overall effectiveness are reported for base rates of .20, .10, .067, and .05. PMID- 10689652 TI - Identification of random responding on the MMPI-A. AB - Although substantial research literature on the effects of random responding on the MMPI-2 exists, there is very limited data available on this issue with the MMPI-A. The purpose of this study was to evaluate the utility of selected MMPI-A validity scales in detecting differences in response patterns between protocols produced by 354 adolescents assessed in clinical settings and a group of 354 randomly produced MMPI-A protocols. Results indicate that MMPI-A validity and basic clinical scales differ significantly between random and clinical groups and that MMPI-A validity Scales F, F1, F2, and VRIN appear to be most useful in correctly identifying protocols from actual clinical participants versus randomly generated response patterns. Findings are discussed in terms of the dramatic effects of the sample base rate for random responding on overall classification accuracy results. Furthermore, it was noted that the optimal cutting scores for MMPI-A Scales F, F1, F2, and VRIN were largely consistent with interpretive recommendations found in the test manual (Butcher et al., 1992) when the relative frequency of random response protocols to clinical protocols was evaluated at a ratio of 1:10. Finally, future recommendations for evaluation of the F1-F2 difference score and the TRIN scale are offered in terms of the most relevant research designs to evaluate these measures. PMID- 10689653 TI - Use of the TAT in the assessment of DSM-IV cluster B personality disorders. AB - The Social Cognition and Object Relations Scale (SCORS), developed by Western, Lohr, Silk, Kerber, and Goodrich (1985), is a diagnostic instrument used to assess an array of psychological functioning by using clinical narratives such as the Thematic Apperception Test (TAT; Murray, 1943) stories. This study investigated the utility of the SCORS to differentiate between Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) antisocial personality disorder (ANPD), borderline personality disorder (BPD), narcissistic personality disorder (NPD), and Cluster C personality disorder (CPD). A sample of 58 patients was separated into four groups: ANPD (n = 9), BPD (n = 21; 18 with a primary BPD diagnosis and 3 with prominent borderline traits who met 4 of the 5 DSM-IV criteria necessary for a BPD diagnosis), NPD (n = 16; 8 with a primary NPD diagnosis and 8 with prominent narcissistic traits who met 4 of the 5 DSM-IV criteria necessary for a NPD diagnosis), and CPD (n = 12). These groups were then compared on the 8 SCORS variables by using 5 TAT cards (1, 2, 3BM, 4, and 13MF). Spearman-Brown correction for 2-way mixed effects model of reliability for the 8 SCORS variables ranged from .70 to .95. The results of categorical and dimensional analyses indicate that (a) SCORS variables can be used to differentiate ANPD, BPD, and NPD; (b) the BPD group scored significantly lower (greater maladjustment) than did the CPD group on certain variables; (c) the BPD group scored significantly lower (greater maladjustment) than did the NPD group on all 8 SCORS variables; (d) the ANPD group scored significantly lower than did the NPD group on certain variables; (e) certain variables were found to be empirically related to the total number of DSM-IV ANPD, BPD, and NPD criteria; and (f) certain variables were found to be empirically related to Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) Personality disorder scales. The results of this study are discussed in terms of clinical utility, conceptual, and theoretical implications. PMID- 10689654 TI - Complexities in complex posttraumatic stress disorder in inpatient women: evidence from cluster analysis of MCMI-III Personality Disorder Scales. AB - Herman's (1992a) clinical formulation of complex posttraumatic stress disorder (PTSD) captures the extensive diagnostic comorbidity seen in patients with a history of repeated interpersonal trauma and severe psychiatric disorders. Yet the sheer breadth of symptoms and personality disturbance encompassed by complex PTSD limits its descriptive usefulness. This study employed cluster analysis of the MCMI-III (Millon, 1994) personality disorder scales to determine whether there is meaningful heterogeneity within a group of 227 severely traumatized women who were treated in a specialized inpatient program. The analysis distinguishes 5 clinically meaningful clusters, which we label alienated, withdrawn, aggressive, suffering, and adaptive. The study examined differences among these 5 personality disorder clusters on the MCMI-III clinical syndrome scales, as well as on the Brief Symptom Inventory (Derogatis, 1993), Dissociative Experiences Scale (E. M. Bernstein & Putnam, 1986), Adult Attachment Scale (Collins & Read, 1990), and Childhood Trauma Questionnaire (D.P. Bernstein, 1995). We present a classification-tree method for determining the cluster membership of new cases and discuss the implications of the findings for diagnostic assessment, treatment, and research. PMID- 10689655 TI - Two-year stability of Psychosis Proneness Scales and their relations to personality disorder traits. AB - Two-year stability of Physical Anhedonia (PhA), Perceptual Aberration (PER), and Magical Ideation (MI) scale scores and their relation to personality disorder traits were examined. Additionally, the effects of a time-lagged (prospective) versus concurrent measurement of psychosis proneness and personality disorder traits were studied to examine the specificity of MI, PER, and PhA. With a non college-student sample (n = 404), stability for PhA was sufficiently high, but for PER and MI, stability was moderate to low. The correlations between personality disorder traits and psychosis proneness scales demonstrate that simultaneous assessment leads to a more nonspecific pattern of associations for MI and PER, although the correlation to schizotypal personality disorder traits were the highest. However, prospectively only MI, but neither PER nor PhA, emerged as a significant predictor for schizotypal and paranoid personality disorder traits in multiple-regression analysis. This suggests that MI may allow for a more specific assessment of psychosis proneness than PER. PMID- 10689656 TI - Problem drinkers: evaluation of a stepped-care approach. AB - The present study evaluated a stepped-care model for the treatment of problem drinkers; those not severely dependent on alcohol. The initial treatment consisted of a motivationally based, four-session outpatient treatment. Based on previous research, treatment nonresponders were defined as having consumed more than 12 drinks per week between the assessment and third session. Six-month follow-up interviews were conducted on three groups of problem drinkers: (1) those who responded to the initial intervention (n = 67); (2) those who did not respond to the initial treatment (n = 36); and (3) those who did not respond to the initial treatment and received a supplemental intervention (n = 33). The last two groups were used to evaluate whether providing treatment nonresponders with an additional "step" would improve treatment outcomes. The primary dependent measures were posttreatment percent days abstinent and posttreatment drinks per drinking day. Results suggested that (1) within treatment drinking can help identify treatment nonresponse in stepped-care models; (2) the supplemental intervention did not influence posttreatment drinking; (3) treatment responders and nonresponders sought additional help at the same rate. The present study is the first study on stepped-care for alcohol treatment and provides a methodology for evaluating stepped interventions. Recommendations for future research in this area include more attention to assessing the needs of treatment nonresponders and help seeking behavior of both responders and nonresponders after an initial intervention. PMID- 10689657 TI - Simultaneous polydrug use among teens: prevalence and predictors. AB - The use of two or more substances in combination, simultaneous polydrug use (SPU), is a particularly dangerous form of drug use that appears to be established by late adolescence. We examined the prevalence of SPU in a diverse sample of 12th graders, and identified risk and protective factors for SPU that are present at 10th grade. We also tested for differences in SPU across race and gender, and explored the basis for observed differences. Our goals were to determine the extent of SPU problems in different groups and how to address these problems. Twenty-nine percent of participants had engaged in SPU in the past year. The best predictors of alcohol/marijuana SPU were a pro-drug environment, pro-drug beliefs, social deviance, and family disruption; only a pro-drug environment was predictive of hard drug SPU. Women were far less likely to combine marijuana and alcohol than were men. Asian Americans were less likely to combine alcohol and marijuana than were other racial groups, apparently due to their advantaged standing on all predictors of this behavior. African Americans were less likely to use hard drugs in combination than were other groups. Overall, polydrug use is a substantial problem for older teens. Broader drug-use prevention programs may be sufficient to address SPU involving gateway drugs, but reducing drug availability appears central to addressing hard drug SPU. PMID- 10689658 TI - The relationship between cannabis use and DSM-IV cannabis abuse and dependence: results from the National Longitudinal Alcohol Epidemiologic Survey. AB - The purpose of-this study was to determine the risk of Diagnostic and Statistical Manual of Mental Disorders--Fourth Edition (DSM-IV) cannabis abuse and dependence at different levels of cannabis use in a nationally representative sample of the U.S. general population. Two separate logistic regression analyses were conducted to determine the association between cannabis use, and abuse and dependence. The risk of cannabis abuse and dependence was found to increase with the frequency of smoking occasions and slightly decreased with age. More severe comorbidity was associated with dependence compared to abuse, suggesting that cannabis might be used to self-medicate major depression. The strength of the association between cannabis use and abuse was also increased as a function of the number of joints smoked among females, but not males. These results were discussed in terms of differential societal reactions, the self-medication hypothesis, and gender biases in diagnosing cannabis abuse. PMID- 10689659 TI - Electrical stimulation therapy in the treatment of cigarette smoking. AB - In this study electrical stimulation therapy (EST) is explored as a possible new treatment for smoking cessation within a randomized controlled trial. The investigation follows reports of several authors that electrical stimulation applied to specific acupuncture points is effective in treating a variety of drug dependencies, including cigarette smoking. Three key features of treatment (electrical stimulation, frequency modulation, and electrode placement), were investigated in a 2 x 2 x 2 factorial design, resulting in eight treatment combinations. Out of 265 smokers recruited into the trial 216 completed the one week treatment. Outcome was assessed in terms of complete abstinence from smoking and symptomatic relief of withdrawal symptoms. Smokers receiving active electrical stimulation obtained higher abstinence rates than those in the inactive groups although the difference did not achieve statistical significance (all active vs. all placebo groups: lambda 1,1(2) = 0.50, p > 0.10, 95% confidence interval = -8.04 to +17.44%; most effective vs. least effective group: lambda 1,1(2) = 3.11, p = 0.08, CI0.95 = -2.2 to +48.8%). The efficacy of electrical stimulation therapy for smoking is not supported. PMID- 10689660 TI - The test-retest reliability of the frequency of multiple drug use in young drug users entering treatment. AB - Assessment of multiple drug use relies primarily on self-report. Several studies support the reliability of client self-reports of drug use but these studies have not involved assessment of the actual frequency of drug use. This test-retest reliability study assessed the frequency of drug use in a clinical sample of 103 multiple drug users, aged 16-25 years. At initial assessment, all participants completed the Drug Use History Form (DUHF) that inquired about the number of drug using days and the daily frequency of use for 13 drug classes during four time intervals. The DUHF was readministered 2-4 weeks later. Reliability was assessed using Intra-class correlations (ICC's). The results indicated that clients do, in general, reliably report both the number of days of use and daily frequencies. The two frequency measures were not highly correlated. Reliability estimates declined over time but most markedly after 90 days, suggesting that assessments of drug use can be reliably extended beyond 30 days. Frequency estimates based solely on the number of days of use of a substance may be unreliable estimates of actual drug consumption, indicating limitations to this commonly used outcome measure. PMID- 10689661 TI - Community reinforcement and family training (CRAFT): engaging unmotivated drug users in treatment. AB - Although motivation for drug abuse treatment is a substantial problem, unilateral intervention through concerned significant others (CSOs) represents a promising method for engaging unmotivated individuals. The Community Reinforcement and Family Training (CRAFT) program, based on principles of reinforcement was developed for this specific purpose. In Phase I, CSOs received the CRAFT intervention, whereby they were taught skills for modifying a loved one's drug using behavior and for enhancing treatment engagement. CSOs were evaluated at 3 and 6 months. In Phase II, engaged drug users received treatment using the Community Reinforcement Approach (CRA). A total of 62 CSOs participated in this evaluation of the effectiveness of CRAFT. CSOs completed, on average, 87% of offered treatment sessions. During the 6-month study period, 74% succeeded in engaging their resistant loved one in treatment. Reported abstinence both from illicit drugs and alcohol increased significantly for drug users engaged in treatment, but not for unengaged cases. All CSOs showed significant reduction in depression, anxiety, anger, and physical symptoms, with average scores dropping into the normal range on all measures. CRAFT provides a promising alternative to confrontational and detachment approaches in counseling CSOs to help their loved ones. PMID- 10689662 TI - Alcohol, intelligence and violent crime in young males. AB - Research has demonstrated a relationship between alcohol and violent behavior, but proof of a causal connection remains elusive. A recent review concluded that the key task that remained was to identify sub-groups of the population for which alcohol promotes violence. Because alcohol might induce violence by causing cognitive disruption (e.g., misunderstood communication), less intelligent persons could be vulnerable because they start out closer to the lower limit of comprehension. Our objective is to investigate the effect of lower intelligence on the alcohol/violence relationship. This analysis uses data from the Buffalo Longitudinal Study of Young Men to investigate this hypothesis. Males, 16 to 19 years of age (N = 596), were selected from Buffalo, NY, by random digit dialing. High-risk males were oversampled. Two interviews were conducted 18 months apart, including drinking, criminal offenses, and psychological traits. Verbal intelligence was measured by the Ammons Quick Test and visual-motor intelligence by the Trail Making Test. An analysis of covariance was conducted with wave 2 average alcohol consumption and both measures of intelligence as independent variables, violent offending as the dependent variable, and race, wave 1 alcohol and wave 1 violence as covariates. Results show a positive main effect of wave 2 alcohol consumption, but also interactions with both verbal and visual-motor intelligence. These interactions indicate that the prevalence of violence increases significantly at low intelligence and high alcohol consumption levels. A parallel analysis with nonviolent offending as the dependent variable failed to find significant interactions. The combination of heavy drinking and lower intelligence is associated with a synergistic surge of violent behavior. PMID- 10689663 TI - Investigation of the tris(trimethoxyphenyl)phosphonium acetyl charged derivatives of peptides by electrospray ionization mass spectrometry and tandem mass spectrometry. AB - Charged derivatives of peptides are useful in obtaining simpler collision activated dissociation (CAD) mass spectra. An N-terminal charge-derivatizing reagent capable of reacting with picomole levels of peptide has been recently reported (Huang et al. Anal. Chem. 1997, 69, 137-144) in the contexts of analyses by fast atom bombardment (FAB) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Electrospray ionization (ESI) mass spectrometric investigation of these tris(trimethoxyphenylphosphonium) acetyl derivatives are described in this article, including studies by in-source fragmentation (ISF) and tandem mass spectrometry (MS/MS). Results from ISF are compared with those from MS/MS. Similarities and differences between ESI-ISF, MALDI-post-source decay (PSD), and FAB-CAD data are presented. Differences in fragmentation of these charged derivatives in the triple quadrupole and ion trap mass spectrometers also are discussed. Application of this derivatizing procedure to tryptic digests and subsequent analysis by liquid chromatography-mass spectrometry is also shown. PMID- 10689664 TI - Unequivocal determination of metal atom oxidation state in naked heme proteins: Fe(III)myoglobin, Fe(III)cytochrome c, Fe(III)cytochrome b5, and Fe(III)cytochrome b5 L47R. AB - Unambiguous determination of metal atom oxidation state in an intact metalloprotein is achieved by matching experimental (electrospray ionization 9.4 tesla Fourier transform ion cyclotron resonance) and theoretical isotopic abundance mass distributions for one or more holoprotein charge states. The ion atom oxidation state is determined unequivocally as Fe(III) for each of four gas phase unhydrated heme proteins electrosprayed from H2O: myoglobin, cytochrome c, cytochrome b5, and cytochrome b5 L47R (i.e., the solution-phase oxidation state is conserved following electrospray to produce gas-phase ions). However, the same Fe(III) oxidation state in all four heme proteins is observed after prior reduction by sodium dithionite to produce Fe(II) heme proteins in solution: thus proving that oxygen was present during the electrospray process. Those results bear directly on the issue of similarity (or lack thereof) of solution-phase and gas-phase protein conformations. Finally, infrared multiphoton irradiation of the gas-phase Fe(III)holoproteins releases Fe(III)heme from each of the noncovalently bound Fe(III)heme proteins (myoglobin, cytochrome b5 and cytochrome b5 L47R), but yields Fe(II)heme from the covalently bound heme in cytochrome c. PMID- 10689665 TI - On-line capillary liquid chromatography tandem mass spectrometry on an ion trap/reflectron time-of-flight mass spectrometer using the sequence tag database search approach for peptide sequencing and protein identification. AB - Capillary high-performance liquid chromatography has been coupled on-line with an ion trap storage/reflectron time-of-flight mass spectrometer to perform tandem mass spectrometry for tryptic peptides. Selection and fragmentation of the precursor ions were performed in a three-dimensional ion trap, and the resulting fragment ions were pulsed out of the trap into a reflectron time-of-flight mass spectrometer for mass analysis. The stored waveform inverse Fourier transform waveform was applied to perform ion selection and an improved tickle voltage optimization scheme was used to generate collision-induced dissociation. Tandem mass spectra of various doubly charged tryptic peptides were investigated where a conspicuous y ion series over a certain mass range defined a partial amino acid sequence. The partial sequence was used to determine the identity of the peptide or even the protein by database search using the sequence tag approach. Several peptides from tryptic digests of horse heart myoglobin and bovine cytochrome c were selected for tandem mass spectrometry (MS/MS) where it was demonstrated that the proteins could be identified based on sequence tags derived from MS/MS spectra. This approach was also utilized to identify protein spots from a two dimensional gel separation of a human esophageal adenocarcinoma cell line. PMID- 10689666 TI - Internal glucose residue loss in protonated O-diglycosyl flavonoids upon low energy collision-induced dissociation. AB - The low-energy collision-induced dissociation of protonated flavonoid O diglycosides, i.e., flavonoid O-rutinosides and O-neohesperidosides, containing different aglycone types has been studied. The results indicate that the unusual [M + H - 162]+ ion formed by internal glucose residue loss, which in a previous study was shown to be a rearrangement ion, is strongly dependent upon the aglycone type. For 7-O-diglycosides, the internal glucose loss is very pronounced for aglycones of the flavanone type, but is completely absent for aglycones of the flavone and flavonol types. Internal glucose residue loss was found to correspond to a minor fragmentation pathway for flavonol 3-O-diglycosides. A plausible mechanism is proposed based on proton mobilization from the aglycone to the disaccharidic part of the flavonoid O-diglycosides which is supported by theoretical calculations and model building. PMID- 10689667 TI - Interpretation of matrix-assisted laser desorption/ionization postsource decay spectra of charge-derivatized peptides: some examples of tris[(2,4,6 trimethoxyphenyl) phosphonium]-tagged proteolytic digestion products of phosphoenolpyruvate carboxykinase. AB - The fragmentation of peptides, to which a positive charge is attached at the N terminus, was studied by matrix-assisted laser desorption/ionization postsource decay mass spectrometry. In these experiments, the tris[(2,4,6 trimethoxyphenyl)phosphonium] acetyl group is covalently attached. The main advantage of this modification is that the resulting spectra are simplified and the fragment ions observed consist predominantly of a(n)-type ions. We report the results for charge-derivatized peptides formed following enzymatic digestion of phosphoenolpyruvate carboxykinase. Specific fragmentation of bonds within aspargine and threonine residues are observed and are discussed. The understanding of the mechanistic aspects of the fragmentation process is essential to formulate a simple and straightforward mass spectrometric strategy for peptide sequencing using these charged derivatives. PMID- 10689668 TI - Matrix-assisted laser desorption/ionization mass spectrometry, enzymatic digestion, and molecular modeling in the study of nonenzymatic glycation of IgG. AB - The glycation-induced functional change of immunoglobulins is of particular interest. The glycation levels of IgG in 10 healthy subjects and 20 diabetic patients with different degrees of metabolic control were studied by matrix assisted laser desorption/ionization (MALDI) mass spectrometry. It reveals the number of glucose molecules that have condensed on the protein, which range from 1 to 5 for healthy subjects, from 5 to 9 for well controlled diabetic patients, and from 10 to 25 for poorly controlled ones. The identification of the most favored glycation sites has been obtained by MALDI analysis of standard and in vitro glycated IgG and plasma protein fraction of a healthy subject after digestion with papain, releasing Fab and Fc fragments of the molecule. Both experiments, as well as molecular modeling of the whole protein, confirm that the most of glucose molecules have condensed on the Fab fragment of IgG, suggesting that the immune deficiency observed in diabetic patients may be explained at the molecular level by a more effective glycation of the Fab fragment, thus inhibiting the process of molecular recognition between antibody and antigen. PMID- 10689669 TI - Tandem time-of-flight experiment for low energy collision studies. AB - We present an experiment adapted to collisional studies of cluster ions based on a laser vaporization setup coupled to a supersonic expansion. The ions are selected in a first time-of-flight, slowed down to the desired energy, and collided in an octopolar guide. The parent and fragment ions are then reaccelerated in order to be mass analyzed in a reflectron time-of-flight. An original method for the extraction of the ion that uses a double voltage pulse, is proposed. The experiment has been applied to collisions of hydrated cobalt ions. An absolute cross section of 17 A2 for the loss of one water molecule by Co(H2O)2+ in collision with neon at a center-of-mass energy of 10 eV, has been determined, with an accuracy of 10%. The threshold for this reaction has been measured at 1.5 eV and is in good agreement with the existing literature (Dalleska et al. J. Am. Chem. Soc. 1994, 116, 3519). Ions that cannot be formed by conventional ligand exchange methods, can also be studied. As an example, the threshold for dehydration of the Co2(H2O)+ ion has been measured at 1.5 +/- 0.2 eV. PMID- 10689670 TI - Gas phase H/D exchange kinetics: DI versus D2O. AB - The gas phase H/D exchange reactions of bradykinin (M + 3H)3+ ions with D2O and DI were monitored in a quadrupole ion trap mass spectrometer. The H/D exchange kinetics of both chemical probes (D2O and DI) indicate the presence of two noninterconverting reactive gas phase ion populations of bradykinin (M + 3H)3+ at room temperature. The H/D exchange involving DI, however, generally proceeds faster than that involving D2O. The rate observations described here can be rationalized on the basis of the "relay mechanism" (see Campbell et al. J. Am. Chem. Soc. 1995, 117, 12840-12854) recently proposed to account for H/D exchange between D2O and gaseous protonated polypeptides. The higher exchange rate with DI is believed to arise primarily as a result of its lower gas-phase acidity relative to that of D2O and, secondarily, as a result of the longer bond length of DI relative to that of OD in D2O. PMID- 10689671 TI - Detection of transthyretin variants using immunoprecipitation and matrix-assisted laser desorption/ionization bioreactive probes: a clinical application of mass spectrometry. AB - In our continuing efforts to develop mass spectrometry-based methods for transthyretin (TTR) variant detection and characterization, we have sought to use matrix-assisted laser desorption/ionization (MALDI) bioreactive probes incorporating immobilized trypsin for screening purposes. These devices show good diagnostic potential as a clinical screening tool to detect amino acid substitutions in TTR. MALDI probes allow the on-probe generation of tryptic digests. The subsequent mass analysis of the on-probe digest yields the peptide map. The inherent advantages of this method include considerably reduced digestion times (minutes vs. hours), absence of autolysis products, minimized sample handling, and hence minimal sample loss. A further advantage is that the opportunity for loss of hydrophobic peptides is reduced because no sample transfer occurs. The method can be applied as a preliminary screen for TTR variants where TTR is isolated from patient serum through immunoprecipitation. This method should also be applicable to other proteins and suitable for automation. PMID- 10689673 TI - [Quality assurance and cost containment--counterproductive or synergistic effects?]. AB - Quality of care and cost considerations are becoming increasingly intertwined. While costs were an insignificant factor in quality management 30 years ago, today the consumption of resources is a measure of the quality of outcome. Newly developed clinical guidelines are intended to reduce the cost of care in the future. However, they entail a significant potential for abuse that can compromise quality. Guidelines are necessary to demonstrate the cost effectiveness of quality control and quality assurance measures. Excessive emphasis on quality assurance may increase the overall consumption of resources. In a managed care setting with budgetary constraints, this can reduce the quality of care. PMID- 10689672 TI - Mass spectrometric analysis of platelet-activating factor after isolation by solid-phase extraction and direct derivatization with pentafluorobenzoic anhydride. AB - Platelet-activating factor is the term used to denote a class of extremely potent lipid mediators that consist predominantly of 1-O-alkyl- and 1-O-acyl-2-acetyl-sn glycero-3-phosphocholines. A method has been devised for rapid isolation of these acetylated phospholipids by solid-phase extraction prior to direct derivatization with pentafluorobenzoic anhydride and analysis by gas chromatography (GC)/electron-capture mass spectrometry. Recovery through the entire method (lipid isolation, derivatization, and purification) typically ranged from 70% to 85%. Using the direct derivatization procedure described here, the practical limit of detection for each of the standard alkyl- and acyl-platelet-activating factor homologs was 1 fmol injected into the GC. Results from the application of the method to the analysis of alkyl and acyl homologs of platelet-activating factor isolated from stimulated human umbilical vein endothelial cells are presented, exhibiting excellent accuracy and precision for a wide range of tissue levels of this class of potent autacoids. PMID- 10689674 TI - ["Fuldaer ventilation surgery"--a surgical concept in severe ventilation disorders of the middle ear]. AB - BACKGROUND: This report presents the long-term results of a special surgical technique in cases of persistent severe dysfunction of the Eustachian tube. PATIENTS AND METHODS: We performed this operation on 16 patients (18 ears) between 1982 and 1997. Almost all patients had undergone previous surgery. This included myringotomies and tube insertions, adenoidectomies, tonsillectomies, myringoplasties and mastoidectomies, sinus surgeries, and an operation on a cleft palate. The concept developed by one coauthor (Draf) combines different methods to improve ventilation of the middle ear and protect the eardrum against partial vacuum. The concept combines mastoidectomy, posterior tympanotomy, and removal of the incus and the head of the malleus with an interposition of the incus. The tympanic membrane is then stabilized with a cartilage-perichondrium graft, and a tube is placed leading from the tympanic cavity to the nose (Wright-tube). A T tube may also be used. The procedure was varied slightly on occasion depending on intraoperative findings. RESULTS: Thirteen patients (15 ears) were available for audiometry at a minimum of 5 months after surgery until 14 years (mean duration of 6.2 years). We compared preoperative and postoperative air-bone gaps. They were measured for the three speech frequencies (500, 1000, and 2000 Hz) at 5 dB intervals. The average preoperative air-bone gap was 25.7 dB versus 18.2 dB postoperatively. This represents an improvement of 7.5 dB. Seven of fifteen ears required revision. Six ears required one revision procedure each (three for cholesteatoma, one for myringitis, one for discharge due to a narrow external auditory meatus, and one for mastoiditis). Another patient required three revision procedures (one for mastoiditis and the other two for cholesteatoma). CONCLUSION: This concept can help prevent cholesteatoma and improve hearing in more than 50 per cent of severe cases of dysfunction of the Eustachian tube. PMID- 10689675 TI - [Use of auditory field measurement in patients with otosclerosis]. AB - BACKGROUND: Middle ear surgery sometimes leads to unpleasant auditory impressions such as distortion or hyperacusis that cannot be detected by conventional audiometric testing. METHOD: Sixty-one patients with conductive hearing loss caused by otosclerosis underwent audiological evaluation, which included a questionnaire followed by testing of the audiometric threshold, speech audiometry, and assessment and quantification of loudness perception with a commercial system (WESTRA). This investigation includes the postoperative measurement of hearing improvement and the patients' subjective impressions regarding hearing increase, distortion of speech, and hyperacusis. RESULTS: Hearing improved in 88% of the patients. A quantification of this hearing increase was possible with pure tone audiometry and the Freiburg speech discrimination test. Reduced hearing threshold and lack of improvement in speech discrimination was confirmed by conventional hearing measurements. However, the presence of hyperacusis and distortion of speech could be determined by conventional audiometry in only 50% of cases. It was interesting to note that the subjects who reported speech distortion and hyperacusis in the questionnaire were identified by their increased loudness perception using the categorical loudness scaling. CONCLUSION: Category loudness scaling appears to be a valuable additional clinical test to characterize postoperative phenomena as distortion of speech and hyperacusis in patients undergoing stapes surgery. PMID- 10689676 TI - [Histopathological studies of intratemporal growth behavior of middle ear carcinoma]. AB - BACKGROUND: Otological hemorrhage otorrhea, and pain are amongst the first clinical signs of the middle ear carcinoma, which is usually diagnosed in advanced stages. Sudden deafness, facial nerve paralysis, and other symptoms of inner ear damage may be observed in the final stage. However, middle ear carcinoma is diagnosed extremely seldom in its early stages. The clinical management of this pathology is based on the knowledge of the tumor's pathways and its anatomic behavior. METHODS: Our study investigated 20 cases of middle ear carcinomas from the Wittmaack temporal bone bank (14 squamous cell carcinomas, 5 adenocarcinomas, and 1 adenoidcystic carcinoma) to analyze the behavior of the tumor growth and its influence on clinical symptoms. The aim was to determine criteria for early clinical diagnosis. RESULTS: The tumor arises in different regions of the temporal bone, and varying symptoms will subsequently reflect its pathway. When the tumor is confined to the middle ear area, its main location is the hypotympanum from which tumor spreads into the eustachian tube and, via infiltration of the adjacent bone structures (anterior wall of the middle ear), into the tensor tympani muscle and the sympathetic plexus of the internal carotid artery. Destruction of the ossicles was observed in the mid-tympanic cavity, and often only a thin layer of fibrous tissue from the Fallopian canal separated the tumor from the facial nerve (this nerve was rarely affected directly). The medial wall (labyrinthine wall) of the tympanic cavity remained intact in the majority of examined cases. The tympanic sinus, the round window niche, and the oval window niche did not show tumor infiltration. In the epitympanum, the tumor grew and infiltrated the adjacent mastoid. Larger tumors affected the internal auditory canal and infiltrated the acoustic nerve and the labyrinth. CONCLUSION: Improving the poor prognosis of middle ear carcinoma requires early diagnosis based on axial computed tomography (CT). Important factors in patient selection include age (50-70 years), sex (mostly women), and especially clinical symptoms (otorrhea, pain, hearing loss). PMID- 10689677 TI - [An anesthesia technique for experimental studies and microsurgical ear interventions in newborn rodents]. AB - BACKGROUND: Experimental investigations on laboratory animals usually require sufficient anesthesia with adequate analgesia and sedation. The technique used should be reliable and easily controllable by the investigator. Here, we present a technique for anesthesia to facilitate invasive and noninvasive investigations in newborn rats and mice. METHODS: Using a custom made breathing mask, anesthesia was induced in these animals with inhalation of gaseous nitrous oxide-oxygen (equal volume at 1 l/min) and halothane (3% by volume). To maintain anesthesia, halothane insufflation was reduced to 1-1.5% by volume. After completion of the experimental procedure, the application of the inhalative gases was determined and substituted by oxygen at 2 l/min. Anesthesia was performed in spontaneously breathing animals. Heart frequency and oxygenation were monitored using a commercially available pulse oximeter. RESULTS: Using the above described technique in neonatal rodents, microsurgery of the ear was performed without signs of pain or major bleeding. Auditory brain stem responses were recorded clearly and reproducible. CONCLUSIONS: This method represents a noninvasive, well tolerated and easy controllable anesthetic procedure which has proven to provide a sufficient and reliable sedation in neonatal rodents for surgical and nonsurgical investigations. PMID- 10689678 TI - [Antibiotic impregnation of cartilage implants: diffusion kinetics of fluoroquinolones]. AB - BACKGROUND: Antibiotic impregnation of cartilage implants may reduce the risk of bacterial infection and subsequent absorption. The aim of this study was to investigate the penetration kinetics of two quinolone antibiotics into fresh cartilage and the concentrations in the core of lyophilized cartilage after rehydration. METHODS: Fresh human costal cartilage was impregnated with ofloxacin and ciprofloxacin (2 mg/ml) for 2, 15 and 90 min. Concentrations were measured in 6 levels (0.5 mm each) from the surface to 3 mm beneath the surface with high performance liquid chromatography (HPLC). Lyophilized human costal cartilage was rehydrated in ofloxacine and ciprofloxacine solutions (2 mg/ml, 0.2 mg/ml and 0.02 mg/ml) for 18 hours and concentrations in the core of the rib segment were measured. RESULTS: Quinolone antibiotics penetrate into cartilage by free diffusion. We found no evidence of significant binding to cartilage. After 2 and 15 min of impregnation, concentrations above the minimal inhibitory concentration (MIC90) for pseudomonas species are found from 0-0.5 mm below the surface of fresh cartilage. After 90 min concentrations above the MIC90 were found 1.0-1.5 mm below surface of the implant. In lyophilized rib grafts which were rehydrated in 1/10 diluted intravenous solutions (0.2 mg/ml), concentrations in the core of the specimen were above MIC90. Differences between the penetration characteristics of ofloxacine and ciprofloxacine were minor. CONCLUSION: Intraoperative impregnation of cartilage implants with ofloxacin or ciprofloxacin probably offers only short-term protection against bacterial infection. Rehydrated rib grafts, however, contain high quinolone concentrations which may be effective even in infected implant beds for several hours. PMID- 10689679 TI - [Value of multimedia educational software in training of the paranasal sinus surgeon]. AB - BACKGROUND: When starting sinus surgery, every surgeon has to pass through an individual learning curve. To avoid complications, costly, time-consuming surveillance is necessary. We wanted to analyse the impact of multimedial teaching software on the learning curve. METHODS: A total of 1104 operations performed by four surgeons were evaluated. The first consecutive 200 operations by each surgeon were analyzed according to their complications. After revaluating the general phases of surgical development, we compared the first 90 operations by another group of four surgeons who had undergone training with the teaching program. RESULTS: Cauterization of the anterior ethmoidal anterior was nearly the same in both groups (10/8). Injuries of the dura dropped from 5 to 2, and periorbital lesions were reduced significantly from 20 to 5 (p < 0.001). CONCLUSIONS: Multimedial learning programs can reduce complications effectively and form a valuable part training. PMID- 10689680 TI - [Endoscopic treatment of iatrogenic esophageal perforation]. AB - BACKGROUND: Esophageal perforations are the most frequent complications of endoscopy of the upper gastrointestinal tract. Life-threatening consequences such as mediastinitis, septic disease, or multiple organ failure are possible. Traditional surgical and conservative methods of treatment should be distinguished. In serious cases, thoracotomy in particular is a high-risk operation. PATIENTS AND RESULTS: This case demonstrates the successful endoscopic treatment of an esophageal perforation with mediastinal empyema by fibrin gluing. CONCLUSIONS: Esophageal perforations up to 20 cm aboral and a maximum diameter of 1.5 cm could be treated by rigid endoscopical fibrin gluing. High-risk patients could be managed effectively avoiding extensive surgery. PMID- 10689681 TI - [Animal experiment study of anastomosis healing after partial resection of the pre-irradiated thoracic esophagus]. AB - BACKGROUND: Multimodal therapeutic concepts in cases of neoplasms of the intestinal tract entail the risk of undesirable complications with respect to healing of wounds and anastomoses. The separate steps of a combined treatment consisting radiation therapy and partial resection of the thoracic esophagus were performed in animal experiments to study the effect of radiation therapy on the healing of anastomoses. METHOD: Adult non-purebred dogs were irradiated in a defined thoracic field with a Betatron (42 MeV) and subsequently underwent esophagectomy. After resection of a 2 cm segment of the esophagus end-to-end anastomosis was performed. Different methods of irradiation and postoperative observation times resulted in a total of 8 groups of 3 animals each. RESULTS: Fractionated irradiation was definitely better tolerated than irradiation with a high single doses. The temporary delay of the anastomotic healing was documented histologically. Only one case of anastomotic leakage occurred, and impaired wound healing was observed in only one animal. CONCLUSION: The mode of irradiation must be regarded as important for the clinical course. Fractionated preoperative irradiation in the area of the thoracic esophagus does not lead to any relevant disturbance of wound and anastomotic healing with meticulous surgical technique and adequate intensive postoperative care. The basic feasibility of surgical therapy combined with preoperative radiotherapy in tumors of the upper digestive tract was confirmed by our experimental work. PMID- 10689682 TI - [Lipoma-like liposarcoma of the tongue]. AB - BACKGROUND: Liposarcomas of the tongue are rare. To date only 7 cases have been reported. PATIENT: We report a new case of a well differentiated lipoma-like liposarcoma of the body of the tongue in a 71-year old woman. Complete uneventful resection was performed. Histologically the tumor consisted of fat cells with occasional lipoblasts and spindle cell areas. There is no sign of recurrence of the tumor 2 years later. RESULTS AND CONCLUSIONS: Liposarcomas of the tongue are rare neoplasms. All cases reported were histologically well differentiated. Liposarcomas have to be considered in the differential diagnosis of solid masses of the tongue. PMID- 10689684 TI - [Tracheal surgery. Tracheotomy]. PMID- 10689683 TI - [Analgesic intolerance and nasal polyps]. PMID- 10689685 TI - [Clinicopathological diagnosis of the diseases in stalk and neurohypophysis]. PMID- 10689686 TI - [Study of the molecular bases of familial amyloidotic polyneuropathy]. PMID- 10689687 TI - [Analysis of segmental motor conduction in the median and the ulnar nerves: comparison between normal and diabetic individuals]. AB - We investigated characteristics of segmental motor conduction in the median and the ulnar nerves. Subjects were 55 individuals with normal conduction of the upper extremity and 71 patients with diabetes mellitus. Mean polyneuropathy index (PNI), which was determined as a mean percentage of the normal for 6 indices concerning to the conduction velocity in the upper limb, was 99.0% in the normal group and 85.6 % in the diabetic group on the mean. In the normal group distal latency was longer in the median nerve than in the ulnar nerve, and the conduction time between Erb's point and the wrist was longer in the ulnar nerve than the median nerve both in men and women. In the diabetic group these differences were accentuated; that means the distal latency was relatively more prolonged in the median nerve and the conduction time between Erb's point and the wrist was much longer in the ulnar nerve. Prolonged distal latency in the median nerve of women and conduction delay between Erb's point and the wrist in the ulnar nerve of men altogether resulted in the gender difference in the median minus ulnar F-wave latency after wrist stimulation in the diabetic group. Carpal tunnel segment of the median nerve and the elbow segment of the ulnar nerve are known to be common entrapment sites. This phenomenon of accentuated conduction delay in these common entrapment sites might be considered as a sort of 'double crush syndrome'. PMID- 10689688 TI - [Effect of number of syllables in word repetition]. AB - We report a case of non-fluent conduction aphasia. The patient was a 59-year-old right-handed male. He suffered from aphasia after a left internal carotid artery occlusion. MRI study revealed subcortical lesions in the left inferior frontal gyrus and cortical lesion in the anterior part of the left insular gyrus and the left postcentral gyrus. The patient showed good comprehension of words and daily conversation but had a common difficulty in the following tasks; naming of pictures, repetition of words, reading of kanji and kana letters. In these tasks, the phonological output of the patient contained many literal paraphasias and there was a tendency that errors appeared more in the posterior portion of a target word. We analyzed the position of errors in the target word on word repetition tasks. The result confirmed the above observation. We speculate that the length of a target word may have played a critical rote in this patient's repetition capability. PMID- 10689689 TI - [A case of cerebellar meningo-encephalitis caused by Epstein-Barr virus(EBV): usefulness of Gd-enhanced MRI for detection of the lesions]. AB - We report a patient with cerebellar meningo-encephalitis by Epstein-Barr virus(EBV) in which the responsible lesions were detected by Gd-enhanced MRI. A 61-year-old woman with a history of liver cirrhosis and diabetes mellitus presented with cerebellar signs such as ataxia of the trunk, bilateral upper and lower extremities and slurred speech two weeks after the acute upper respiratory inflammation for several days. Serum IgM antibody(Ab) to EBV viral capsid antigen(VCA) was 1:10, Ab to EBV(VCA) IgG was 1:1280, Ab to early antigen diffuse and restricted (EADR) IgG was 1:40, Ab to EBV nuclear antigen (EBNA) was 1:80. Other viral antibody titers were not elevated significantly in serum. Cerebrospinal fluid (CSF) pressure was 195 mmH2O, containing 464 cells/mm3, protein 68 mg/dl and glucose 43 mg/dl. Only CFS Ab to EBV(VCA) IgG elevated significantly (1:16). In acute phase plain MRI was normal except for swelling of the cerebellar hemispheres while Gd-enhanced MRI showed a leptomeningeal enhancement of bilateral cerebellar hemispheres and of vermis disappeared within one month. A homogeneously enhanced lesion in the left dentate nucleus appeared one month after the onset of illness. This lesion had been detected on Gd enhanced MRI for three months after clinical symptoms were improved. No abnormal finding was shown in the supratentorial region during the whole clinical course. In the literature, EBV encephalitis has a wide range of MR findings which may vary in a short period. We emphasize that frequent MR examinations including Gd enhanced MRI is useful to evaluate inflammatory or demyelinating diseases in the posterior fossa. PMID- 10689690 TI - [A case of herpes zoster encephalitis with Ramsay-Hunt syndrome, herpes zoster generalisatus and acute pancreatitis]. AB - We describe here a 71-year-old man who had herpes zoster encephalitis. He developed high fever, headache and disturbance of consciousness on 1st, May, 1998. On admission, neurological examination revealed disturbance of consciousness with restlessness and meningeal signs. Brain MRI (T 1 and T 2 weighted images) demonstrated high signal lesions in the left temporal lobe and cerebellar vermis. VSV encephalitis was diagnosed based on CSF pleocytosis, high serum and CSF titers of VZV antibody and EEG abnormality. During hospitalization, Ramsay-Hunt syndrome, herpes zoster generalisatus and acute pancreatitis developed. To our knowledge, the characteristic combination of the clinical signs in this case is very rare. We discussed the pathogenic mechanisms of these conditions, and this case was considered to have VZV encephalitis, and to be associated with right facial nerve palsy and pancreatitis, in spite of the absence of immunological deficiency. PMID- 10689691 TI - [Transient improvement in motor function and hemineglect by vestibular stimulation in a patient with right middle cerebral artery embolism]. AB - A 62-year-old Japanese man presented left hemiparesis and left visuospatial hemineglect following a right hemispheric stroke. His CTs and MRIs of the brain revealed a large embolic infarction of the middle cerebral artery territory. A month after the cerebrovascular event, his weakness of the left lower limb almost recovered fully. However, his upper limb motor function was still disabled; in particular, his ability of finger flexion in the left hand was almost lost. Then, vestibular stimulation using either a cold caloric stimulation to the left ear or a warm caloric stimulation to the right ear was performed, and the effect on the hemineglect symptoms were assessed by a line bisection task. After vestibular stimulation, not only his hemineglect symptoms but also his motor functions of left upper limb transiently improved; he became able to make a fist. The improvement of his hemineglect symptoms was obtained by vestibular stimulation using either a cold or a warm caloric stimulation. However, the effect on the motor function was obtained only by the cold caloric stimulation applied to the left ear. Based on the effect of the vestibular stimulation, we postulates that the impairment of the motor function in the present patient is not only a paresis caused by the pyramidal tract lesion but also symptoms related to the hemineglect syndrome. PMID- 10689692 TI - [A case of rhabdomyolysis with water intoxication confirmed by muscle biopsy]. AB - A 32-year-old woman with chronic schizophrenia who took 8-10 liters of water for three years due to thirsty, admitted to our hospital because of convulsion and muscle weakness. Neurological finding on admission showed a mild disturbance of consciousness, moderate proximal muscle weakness, and muscle pain. Laboratory examination revealed marked serum hyponatremia(102 mEq/l) and high value of creatin kinase (1,259 IU/l). The level of creatin kinase reached a peak(39,700 IU/l) at the 5th hospital day. An analysis of the muscle biopsy specimen showed necrotic muscle fibers and opaque fibers, that was compatible with rhabdomyolysis. T 2 weighted magnetic resonance imaging of the brain showed a transient high signals in bilateral putamen but not in pons. She was diagnosed to have rhabdomyolysis due to water intoxication. The present case is the first rhabdomyolysis in Japan that was confirmed by muscle biopsy at an acute stage of water intoxication related with schizophrenia. PMID- 10689693 TI - [A case of Wernicke-Korsakoff syndrome with dramatic improvement in consciousness immediately after intravenous infusion of thiamine]. AB - A 68-year-old man was hospitalized on March 4, 1998 for disturbances in consciousness. In 1995, he had received proximal subtotal gastrectomy and reconstructive surgery of the jejunal interposition for gastric cancer. Thereafter he had been taking enough food without the habit of taking liquor. In October 1997, his short term memory was becoming gradually worse. On February 12, 1998, he suffered from numbness in the feet, and then dysphagia, unsteady gait, and diplopia developed gradually. On February 26, brain MRI showed no abnormalities. On March 3, he had a fever of 38.5 degrees C and his consciousness became unclear. Neurological examination revealed semi-coma, total ophthalmoplegia, and absence of doll's eye movement. Deep tendon reflexes were absent. The serum thiamine level was 9 ng/ml (normal range: 20-50). Brain MRI demonstrated symmetrical high intensity lesions in the periaqueductal area of the midbrain, dorsomedial nuclei of bilateral thalami, and vestibular nuclei. About 30 seconds after intravenous infusion of thiamine, his consciousness improved dramatically, but returned to semi-coma after about two minutes. Wernicke Korsakoff syndrome usually occurs acutely. In the present case, however, the disease showed slow onset, chronic progression, and then rapid worsening after fever. Reconstructive surgery of the jejunal interposition might have caused the slow onset of Wernicke-Korsakoff syndrome, and fever might have facilitated the rapid progression of the disease. An immediate high concentration of thiamine modifies the kinetics of acetylcholine receptor ion channels, thereby maintaining wakefulness, and the level of consciousness may change dramatically. PMID- 10689694 TI - [An autopsy case of catastrophic antiphospholipid syndrome presenting with recurrent multiple cerebral infarction associated with lung cancer]. AB - We reported an autopsy case of cerebral infarction with primary lung cancer. The patient was a 50-year-old man. Despite having been treated with warfarin potassium and ticlopidine hydrochloride, he relapsed cerebral infarction. His laboratory data on admission showed that lupus anticoagulant was positive, together with a high value of beta-thromboglobulin, thrombin-antithrombin III complex, markers of platelet and coagulation activation, CEA and CA 19-9. The autopsy finding revealed a primary papillary adenocarcinoma in the right lower lung, multiple cerebral infarction, renal infarction, pulmonary infarction and splenic infarction. The atherosclerotic changes were mild in the whole tissues and findings of vasculitis were not observed. Recurrence of cerebral infarction was effectively suppressed with the addition of steroid therapy to antithrombotic therapy. This case was considered as catastrophic antiphospholipid syndrome. It is necessary to differentiate antiphospholipid syndrome in case of the abnormal coagulation and fibrinolytic factors with recurrent cerebral infarction. Moreover, systemic examinations are important, because malignant tumor may exist on the background of the case. PMID- 10689695 TI - [Visual functional MRI: a case of intracranial meningioma with hemianopsia]. PMID- 10689696 TI - [Metastatic bone tumor of the clivus]. PMID- 10689698 TI - A serological survey of bovine babesiosis in northern and eastern Zimbabwe. AB - The geographical distribution of Babesia bovis and Babesia bigemina antibodies in communal herds in northern and eastern Zimbabwe was determined using the ELISA technique. The animals in different herds in the study region had different levels of natural exposure to B. bovis (mean 32%, range 0-79%) and B. bigemina (mean 52%, range 5-92%) infections. The majority of herds (90%) were endemically unstable for B. bigemina and 62% were unstable for B. bovis. Natural region 5 and Manicaland province had the highest seroprevalence of B. bovis infection, while natural region 5 and Masvingo province had the highest seroprevalence of B. bigemina infection. PMID- 10689697 TI - [An autopsy case of bilateral carotid artery occlusion with repetitive epilepsy and brain atrophy in a senile patient]. AB - A 67-year-old man was referred to us for tonic-clonic convulsions. A review of his history revealed that he had been hospitalized for loss of consciousness, hypotension, and suspected apoplexy at age 67. He had experienced prior tonic clonic convulsions at age 72 and age 74. He had malaria and tuberculosis in his history but had been otherwise generally well. Physical examination was normal, and his blood pressure was 100/80 mmHg. Laboratory findings were normal except alcalinephosphatase (292 U/l) and gamma-glutamyl transpeptidase(60 U/l). Neurological examination showed alert consciousness, mild upper gaze palsy, slight right-side hemiparesis and left Babinski signs was present. Cranial magnetic resonance imaging showed no abnormality, but cerebral angiography revealed bilateral carotid artery occlusion. There were abundant leptomeningeal anastomoses, and the posterior communicating artery was supplied by the left vertebral artery. Electroencephalography showed a spike wave in the temporal lobe and rebuild-up phenomenon in the right hemisphere. Brain atrophy in the anterior and temporal lobes progressed, and the patient experienced gradual disorientation, delirium and hypobulia. He was eventually bedridden. He also demonstrated repetitive tonic-clonic convulsions. After one convulsion, he remained unconscious and died of pneumonia. Autopsy revealed thickening of the intima and internal elastic lamina in the occluded internal carotid artery. The anterior and middle cerebral arteries showed the same pathological changes. Multiple small infarctions restricted to grey matter were present in the cerebral cortex and may have caused the progressive brain atrophy. There was no myelin pallor in the white matter of the cerebrum. Atherosclerotic changes, senile plaque, and neurofibrillary tangles were seen but were within normal limits. These pathological findings were strongly suggestive of moyamoya disease. PMID- 10689699 TI - The production and evaluation of Pasteurella haemolytica leukotoxin in the supernatant of submerged cultures in fermenters. AB - The optimal production of P. haemolytica leukotoxin in the culture supernatant of a fluid medium is dependent on a number of factors. The leukotoxin has to be produced by using a strain that is known for its ability to produce high quantities of leukotoxin, inoculated into the most suitable type of medium at the correct culture density containing the necessary supplements and harvested after a certain growth period. The volume in which it is produced may also have an influence. Two different procedures are described to produce the leukotoxin in 5 to 15-l quantities in RPMI 1640 medium. The first method used to produce leukotoxin is one that has been repeatedly described since the presence of the leukotoxin was first established in 1978. Using this method seven batches of leukotoxin were produced in litre quantities with leukotoxin activity ranging from 23-67 u/ml. The seed culture inoculum is prepared in brain heart infusion broth, which is centrifuged before the organisms are inoculated into RPMI 1640 medium containing 3.5% foetal calf serum and incubated for only 1 h in a fermenter, after, which the leukotoxin is harvested. An improved alternative method was devised which yielded higher levels of leukotoxin activity by utilising the ability of the P. haemolytica organisms to grow and produce leukotoxin during the logarithmic growth phase in a fermenter. A seed culture harvested in the log phase was prepared in brain heart infusion broth by means of a series of cultures and inoculated into RPMI 1640 containing 3.5% foetal calf serum. Three hours of active growth were allowed during which the leukotoxin was measured by its biological activity and an ELISA assay, and the increase in cell mass by means of the optical density every 30 min. The average leukotoxin biological activity measured 260 u/ml and by means of the ELISA test the leukotoxin concentration measured 315 u/l which is a substantial increase in leukotoxin production. In comparison the average optical density only measured 0.469 at 650 nm. Previous findings were substantiated that the highest cell density was not reflected in the highest leukotoxin activity. It is possible to induce high levels of leukotoxin secretion in submerged cultures with RPMI 1640 medium containing foetal calf serum in the controlled environment of a fermenter in large enough quantities for use as a vaccine by the improved preparation of the seed culture inoculum. PMID- 10689700 TI - Anthelmintic resistance in South Africa: surveys indicate an extremely serious situation in sheep and goat farming. AB - Surveys to determine the prevalence and degree of resistance of Haemonchus spp. of sheep and goats to the available anthelmintics in South Africa indicate that small ruminant production is entering a crisis situation. Three surveys employing the faecal egg count reduction (FECR) test to determine resistance were conducted in some of the main sheep-producing areas in the summer rainfall region of South Africa, where H. contortus is the principal worm species in sheep. After analyzing the data recorded in the surveys by six different methods, including the RESO test at two different levels of confidence, the results obtained in the least stringent one (geometric mean reduction of the worm egg counts of drenched, vs untreated group of sheep) are reported in this paper, so that if any bias was obtained it would be in the favour of the anthelmintic. In Mpumalanga and KwaZulu Natal there was anthelmintic resistance in Haemonchus spp. on all the 52 farms surveyed. Sixteen percent of the strains of H. contortus were < 60% susceptible to three of the four anthelmintics tested, and 8% of the strains were < 40% susceptible to all four of the anthelmintics. FECR tests of sheep in six localities in the Lebowa district of Northern Province indicated that even in previously disadvantaged communities where anthelmintic treatment is less intensive, anthelmintic resistance is developing, and is possibly at the level at which the situation on commercial sheep and goat farms in South Africa was 25 years ago. From the data it appears that the level of anthelmintic resistance of H. contortus in South Africa is possibly the highest that has so far been recorded in the world and that strains of it are emerging that may soon not be controllable by treatment with any of the existing anthelmintics. Farmers in the summer rainfall region, if not the whole country, must be alerted to the immediate need for testing the parasite burdens of their sheep for susceptibility to preparations in all four groups of anthelmintic compounds currently available. Alternative methods of integrated worm control, including biological, must be sought and implemented with urgency, to reduce further selection for resistance and to induce reversion of the resistance that has already developed. PMID- 10689702 TI - Evaluation of coloured targets for the attraction of Glossina brevipalpis and Glossina austeni (Diptera: Glossinidae) in South Africa. AB - Studies on the attractiveness of various coloured targets for Glossina brevipalpis and G. austeni in South Africa showed black and pthalogen blue (p.blue) combinations to be the most effective for both species. A 2 m wide (all targets 1 m high) black/p.blue/black (colour ratio 1:2:1) conformation caught nearly three times more G. brevipalpis and nearly five times more G. austeni than a 1.5 m wide black standard control target. For G. brevipalpis the black/p.blue/black (1:2:1) target should be at least 2 m wide in order to increase catches significantly while a 1.5-2.0 m wide target is optimal for G. austeni. The p.blue section of a 2 m black/p.blue/black target should not make up less than 20% of the total target width for either species. The most effective combination of practical target sizes and colour ratios were a 1.75 m wide black/p.blue/black (1:1.5:1) or 2 m wide target (1.5:1:1.5). Between 61-95% of G. brevipalpis and 34-90% of G. austeni that were attracted, settled first on the black section of black/p.blue targets (> 1 m wide). Further studies revealed that for G. brevipalpis only the black parts of the 2 m wide target need to be treated with insecticide, while the entire 1.75 m wide target should be treated. For G. austeni the total width of either target should be treated with insecticide since this species readily settles on both blue and black. PMID- 10689701 TI - A comparison of the infectivity of cryopreserved versus unfrozen infective larvae of Haemonchus contortus, Trichostrongylus colubriformis and Trichostrongylus axei. Results of the Onderstepoort Veterinary Institute and collaborators from 1977 to the present. AB - The infectivity for sheep of cryopreserved infective larvae (L3) of various strains of Haemonchus contortus, Trichostrongylus colubriformis and Trichostrongylus axei is compared using previously published results of trials conducted at the Onderstepoort Veterinary Institute laboratories, and of collaborators. The means and ranges of development were similar for both frozen and unfrozen larvae of two of the three worm species reviewed. A mean of 33.4% (range, 12.7-63.0%) of cryopreserved H. contortus L3 developed, compared to a mean of 43.7% (range 2.4-78.7%) of unfrozen L3 of this worm species. The corresponding values for T. colubriformis were 33.0% (range 10.3-62.7%), and 33.5% (range 8.3-52.2%), respectively. In the case of T. axei, the development of the cryopreserved L3 (tested in only three trials) was markedly lower than that of unfrozen L3 in the single trial in which the latter was evaluated. It is concluded that development of cryopreserved L3 is probably similar to that of unfrozen L3 and that, for several reasons, maintaining nematode larvae in the frozen state in liquid nitrogen is a much superior method to that of one which entails cycling worm strains continually in their final hosts. PMID- 10689703 TI - Evaluation of conventional odour attractants for Glossina brevipalpis and Glossina austeni (Diptera: Glossinidae) in South Africa. AB - The components of the synthetic ox-odour used in Zimbabwe against Glossina pallidipes and G. m. morsitans were evaluated for the attraction of G. brevipalpis and G. austeni in South Africa. The Zim babwe mixture (Zim-mix), which consisted or acetone and a 1:4:8 mixture of 3-n-propyl phenol, 4-methyl phenol and 1-octen-3-ol, increased the catches of G. brevipalpis by c. 2.1-4.4 times compared to when no odours were used. One of the odour components, namely 3 n-propyl phenol, did not significantly increase the size of the catches. Acetone was an essential component for G. brevipalpis, especially during the warm and wet season when it acted synergistically with high doses of 1-octen-3-ol and 4-methyl phenol. The most attractive odour combination for G. brevipalpis was 1-octen-3-ol released at 2.3-9.1 mg/h with 4-methyl phenol at c. 15.5 mg/h and acetone at c. 350 mg/h. This combination increased the catches by another 2.3-2.8 times when compared to the Zim-mix and 10.1-12.3 times compared to 'no odour'. None of the odour components was attractive for G. austeni. None of the components was repellent for either species. PMID- 10689704 TI - An anatomical study of the respiratory air sacs in ostriches. AB - An accurate description of the number, location and relative position of the air sacs and their diverticula in the ostrich is essential for a better understanding of the pathogenesis of air sacculitis in this bird. The air sacs were studied in ten ostriches of varying ages by latex or silicone casting of the respiratory tract and dissection. Results revealed that the air sacs of the ostrich conform to the general pattern in birds. Cervical, lateral and medial clavicular, cranial and caudal thoracic, and abdominal air sacs are present. The left and right medial clavicular air sacs fuse with each other ventrally to the trachea to form a single, median compartment. A unique, large gastric diverticulum which covers the caudal aspects of the proventriculus and gizzard originates from the median compartment of the clavicular air sac. The lateral clavicular air sacs and their diverticula are similar to those of other bird species, with the exception that humeral diverticula are absent. Both abdominal air sacs are relatively small, with the left sac being the larger. Perirenal and femoral diverticula, similar to those found in other bird species, are present. However, the entire femur is aerated by the femoral diverticulum which also forms a large, subcutaneous division caudally and caudo-laterally to the femur. The presence of this subcutaneous part has practical implications for injury and intramuscular injections. The number and location of ostia connecting the air sacs to the bronchial tree are generally similar to those reported in other bird species. PMID- 10689705 TI - Evaluation of a proposed odour-baited target to control the tsetse flies Glossina brevipalpis and Glossina austeni (Diptera: Glossinidae) in South Africa. AB - The most effective odour attractant for G. brevipalpis Newstead, namely a combination of octenol released at c. 9.1 mg/h, 4-methyl phenol released at c. 15.5 mg/h and acetone released at c. 350 mg/h, when used together with the smallest recommended colour target (as determined in previous studies), namely a 1.75 m wide x 1 m high black/pthalogen-blue/black target, was evaluated for the control of G. brevipalpis and G. austeni Newstead. This combination increased the catches of G. brevipalpis by 3.5 fold when compared to the number of those caught on a 1.5 m wide x 1 m high black target baited with a synthetic ox-odour as was used in a trial to control this species in the Hluhluwe-Umfolozi Game Reserve in 1992. There was an indication that odour (olfaction) plays a far more important role in attracting G. brevipalpis than does colour (vision). For G. austeni visual attraction appears to play the major role as the odours used were relatively unattractive to them. The odour-baited target should, however, attract G. austeni in sufficient numbers (visually) to achieve control to the fly. PMID- 10689706 TI - Detection of Mycoplasma gallisepticum and Mycoplasma synoviae antibodies in the sera of indigenous chickens by rapid serum agglutination test at Mmopane, Gaborone, Botswana. AB - The mean flock size was ten chickens per rural farmer. Antibodies to Mycoplasma gallisepticum and Mycoplasma synoviae were detected in 57.88% and 67.33% of the chicken sera respectively. PMID- 10689707 TI - The helminths of ranch calves in the North-eastern Mountain Grassland of South Africa. AB - The cumulative total helminth parasite burdens of ranch calves during their first seven months of life on the North-eastern Mountain Grassland of South Africa were determined during two consecutive years. Trichostrongylus axei was the most abundant nematode parasite followed by Cooperia spp. and Ostertagia ostertagi. Haemonchus spp. occurred in relatively low numbers and its development was significantly inhibited. The total helminth parasite burdens of the calves ranged from 681 to 7,269 with a mean of 4,405. PMID- 10689708 TI - DNA ploidy and prognosis of neuroblastoma. PMID- 10689709 TI - "To the heart of the matter...". PMID- 10689710 TI - Lymphokines post autologous peripheral blood stem cell transplantation in children. PMID- 10689711 TI - Pediatric hematology and oncology at the University Children's Hospital, Basel, Switzerland. AB - The department offers the management and research of benign and malignant pediatric hematology and oncology at a comparable European standard. It serves as a referring department for BMT, thoracic and visceral pediatric surgery, and pediatric orthopedic surgery and is equipped with all facilities of modern oncology and surgery. Psychooncology is a continuing interest at our department and may help children and their families to better understand and accept their situation and to receive the support necessary for coping with the threat of malignant diseases. Our department aims at practicing a highly sophisticated communication culture essential for a multidisciplinary approach to the child with cancer, representing a central condition for the success of the treatment of malignant diseases. PMID- 10689712 TI - Persistent altered spermatogenesis in long-term childhood cancer survivors. AB - This study evaluated male gonadal function in long-term survivors of childhood cancer and assessed the suitability of offering sperm analysis to all those patients independently of the diagnosis and treatment received. A total of 43 survivors of acute lymphoblastic leukemia (21), acute myeloid leukemia (1), neuroblastoma (8), ganglioneuroblastoma (1), ganglioneuroma (2), Wilms' tumor (9), and mesoblastic nephroma (1) underwent sperm analysis at a mean age of 20.2 years, after a mean time off treatment of 13.6 years. Eight of the patients (19%) were azoospermic, 2 (5%) were severely oligo-asthenozoospermic, and only 16 (37%) were normozoospermic. A control group of healthy volunteers aged < or = 30 years included no azoospermic subjects, 7% severely oligo-asthenozoospermic, and 67% normozoospermic. Comparisons were also made with patients treated at our Human Reproductive Unit aged < or = 30 years (n = 373) whose percentages for the above parameters were 4, 9, and 42%, respectively. Cumulated cyclophosphamide dose and basal follicle-stimulating hormone (FSH) levels were identified as independent factors associated with azoospermia or severe oligo-asthenozoospermia. Azoospermic and severely oligo-asthenozoospermic survivors had significantly smaller mean testicular volume and higher basal FSH levels than the other survivors, but small testicles (sum of both testicular volume < or = 20 mL) and/or abnormally high basal FSH (> 10 mIU/mL) were present in only half of the azoospermic survivors. Male long-term survivors of childhood cancer constitute a high-risk subpopulation for altered sperm analysis. It seems justified to offer sperm analysis to all long-term survivors. PMID- 10689713 TI - Clinical ineffectiveness of IL-2 and/or IFN alpha administration after autologous PBSC transplantation in pediatric oncological patients. AB - Clinical impact of s.c. administration of IL-2 and/or IFN alpha was studied in 23 pediatric patients with Hodgkin lymphoma (IFN alpha group) and sarcoma, non Hodgkin lymphoma, peripheral neuroepitelioma, neuroblastoma, and embryonic carcinoma (IL-2 + IFN alpha group) after autologous PBSC transplantation. Expression of CD3, CD4, CD8, CD25, CD38, CD56, CD71, CD122, and HLA-DR antigens, serum level of the soluble IL-2R alpha, and NK activity against K562 cell line were evaluated in 11 patients representative for both types of immunotherapy. T and, more markedly, NK cell proliferation, induction of activation markers on the surface of T and NK subsets, and elevation of sIL-2R alpha concentrations were seen in the IL-2 + IFN alpha subgroup. In the IFN alpha subgroup, the total number of lymphocytes and expression of activation markers remained unchanged, but the number of CD8+ T cells increased at the expense of CD4+ T and NK cells during the therapy. Cytotoxic activity against K562 cells was not influenced by the immunotherapy in either subgroup. No significant clinical benefit of the immunotherapy was seen in these patients compared to 27 control patients with relevant diagnoses who did not receive immunotherapy. PMID- 10689714 TI - DNA content and proliferative activity in children with neuroblastoma. AB - Many studies have reported that neuroblastoma patients with aneuploid DNA content and a low cellular proliferative activity have better outcome; other studies have reported contradictory results. Formalin-fixed, paraffin-embedded archival pretreatment specimens of 56 neuroblastomas were studied. Thick sections from paraffin blocks were deparaffinized, and rehydrated. Following enzymatic digestion and filtration, cellular suspensions were analyzed by flow cytometry. Six tumors were aneuploid (13.3%) and 39 samples were diploid (86.7%). S-phase fraction (SPF) ranged from 1 to 78% with a median of 31%. DNA ploidy and proliferative activity results showed no correlation with the prognostic variables. There was no significant difference between the 5-year overall and event-free survival rates of the aneuploid and the diploid neuroblastomas or between the neuroblastomas with a high and low proliferative activity. The results revealed the prognostic significance of neither DNA ploidy nor the cellular proliferative activity in neuroblastoma in contrast to other studies. PMID- 10689715 TI - Use of alternative therapy among pediatric oncology patients in Taiwan. AB - Both alternative medicine and western medicine have been commonly used to treat pediatric cancer patients in Taiwan. Each has its own intrinsic strengths and weaknesses and they can be complementary. Little is known about medical help seeking behaviors of parents of pediatric cancer patients, especially those related to alternative therapies. This study investigated the extent and parental expectations on use of alternative therapies. All primary caregivers of 63 eligible patients were interviewed. Use of alternative therapies, regardless of education level or social status of their families, is prevalent (n = 46, 73%) in Taiwan. Commonly used alternative therapies included, in order of popularity, formulated functional food (n = 22, 48%), temple worship/shamanism (n = 19, 40%), traditional Chinese medicine (n = 9, 20%), secret recipes/herbs (n = 13, 28%), and diet supplements (n = 9, 19%). Such practices generally occur without medical guidance from oncologists, largely because of poor interactions between parents and oncologists. Future efforts should be made to encourage both parents and oncologists to discuss this issue. Nurses may serve as mediators by developing mutual trust and a sharing relationship between these groups. PMID- 10689716 TI - Changes in the policies of the department of hematology, 1982-1998, designed to promote the mental health of children with leukemia and enhance their quality of life. AB - A perceived personal control (PPC) preventive intervention model that had earlier received empirical and theoretical verification was applied to a population of pediatric leukemia patients to promote their mental health and enhance their quality of life. The PPC model entails intervention on two complementary levels. On the personal interaction level, preventive intervention is administered by a network of natural and organized support systems, while the social action level leads to the introduction of changes in policies, structures, allocation of resources, and services. Preventive intervention in both domains is discussed with respect to changes in policies introduced between 1982 and 1998 in the hematology department of Rambam Medical Center in Haifa, Israel, as well as in other relevant departments catering to children with leukemia, to foster the positive mental health of these children. PMID- 10689717 TI - A longitudinal study of cardiac function in children with cancer over 40 months. AB - A previous study demonstrated impaired systolic function in 29% of patients treated with anthracycline as part of their therapy for malignant disease. A follow-up echocardiographic study was performed to determine whether there had been further deterioration of cardiac function. At least 40 months after the first study, those patients in whom abnormal systolic function had been detected and who had not received further anthracycline were studied by echocardiography using the same protocol as the initial study (group A). A second group of pediatric oncology patients who had not been given anthracycline but who had previously had cardiac assessment was selected as a control group (group N). The age and sex distributions of the two groups were comparable. Group A comprised 29 patients assessed on 2 occasions at mean times of 46 months and 89 months from the last dose of anthracycline. The mean dose of anthracycline received was 233 mg/m2 (range 20-400). Nine of 16 patients and 4 of 5 patients who had abnormal ejection fraction (EF) and fractional shortening (FS) at first assessment had normal EF and FS at the second assessment. There were no significant changes in EF, FS, and left ventricular wall stress (LVWS) between the two examinations. In group N, 20 patients were assessed after a mean interval of 43 months. There were no significant changes in EF, FS, or LVWS between the two examinations. At the first but not the second examination there were significant differences in the left ventricular internal diameters, EF, FS, and LVWS between group A and group N. Mildly abnormal cardiac indices detected in children after cessation of treatment with anthracycline did not deteriorate in 3 to 4 years follow-up. A longer cardiac follow-up study is indicated to assess the late outcome. PMID- 10689718 TI - Pediatric thyroid carcinoma: 22 years of experience at the Northern Israel Oncology Center (1973-1995). AB - Over the past 22 years, 16 children with thyroid carcinoma were referred to the Northern Israel Oncology Center. All patients had undergone surgical procedures, either total or subtotal thyroidectomy, and 7 patients had undergone cervical lymph node dissections. Postoperatively, 5 patients underwent thyroid ablation with radioactive 131I as first treatment. Two patients received postoperative external radiation therapy to a field encompassing the cervical region, superior mediastinum, and both supraclavicular grooves. After a median follow-up of 60 months (range, 5-169 months), all patients are alive with no evidence of recurrent disease. Two patients who had recurrences, one in the submaxillary lymph nodes and one in the lungs, were salvaged successfully with retreatment with 131I therapy. No severe acute or long-term side effects were exhibited. The long-term results of treatment of pediatric thyroid carcinoma are excellent, but there remains disagreement over the extent of surgical and postsurgical treatment required. PMID- 10689719 TI - Antibiotic sequential therapy for febrile neutropenia in pediatric patients with malignancy. AB - Children suffering malignant diseases can experience phases of bone marrow depression during intensive chemotherapy. The influence of antibiotic sequence therapy on the course of diseases was examined in 41 pediatric patients with malignant diseases. Inclusion criteria were neutropenia (ANC < 500/microL), rectal body temperature over 38.5 degrees C, and increased C-reactive protein (CRP, cutoff > 5.0 mg/L). The first stage of therapy comprised the following antibiotics: piperacillin, teicoplanin, and gentamicin. In stage 2 imipenem, teicoplanin, and tobramycin were administered. Fluconazole was the antifungal drug of choice in stages 1 and 2. In the first level of antibiotic therapy 68% of the patients showed a positive reaction. The C-reactive protein was a sensitive parameter, which significantly decreased with 3 days of therapy. A total of 72% of the bacteriological smears were sterile. All patients survived the septic phase. PMID- 10689720 TI - Neutropenic enterocolitis in children with acute lymphoblastic leukemia. AB - Neutropenic enterocolitis is an acute, life-threatening inflammation of the small and large bowel, often seen in children with malignancies during periods of prolonged or severe neutropenia. During the period 1990-1995, 180 children were treated at the authors' center for acute lymphoblastic leukemia using a standard chemotherapy protocol. Among them, 11 children (6.1%) aged 4 to 12 years, were diagnosed clinically to have neutropenic enterocolitis. Eight had severe neutropenia (absolute neutrophil count < 10(8)/L and 5 had prolonged neutropenia (> 7 days duration). The symptoms included diffuse abdominal pain (10 children), oral mucositis (7), hematochezia (7), diarrhea (6), hematemesis (5), and right lower quadrant tenderness (4). Three children had radiological evidence of free intraperitoneal gas and an additional 3 children were found on surgical exploration to have cecal perforation. Laparotomy was performed on 8 children (73%), 4 of whom survived. Among the 3 children managed conservatively, 1 died awaiting surgical exploration, while the other 2 did well. The overall survival was 55%. The authors recommend an approach to management that respects the heterogeneity of the disease. PMID- 10689721 TI - Testicular germ cell tumors in prepubertal children. AB - Pediatric testicular germ cell tumors are rare. Fifteen children, all less than 5 years of age, were evaluated and treated during February 1987 to July 1996. The median age was 18 months (range, 4-60 months). All were staged according to the Pediatric Oncology Group/Children's Cancer Study Group staging system. Seven patients had stage III disease. Histologically, 9 patients had pure endodermal sinus tumor, 1 had endodermal sinus tumor with embryonal carcinoma, 1 had embryonal carcinoma alone, 2 had immature teratoma, and 2 had mature teratoma. Six children were kept on surveillance. All others received chemotherapy with cisplatin, bleomycin, and vinblastine. The 10-year actuarial overall survival rate was 86.7%. PMID- 10689722 TI - Pneumomediastinum, subcutaneous emphysema, and pulmonary fibrosis in a patient with idiopathic pneumonia syndrome after bone marrow transplantation. AB - An adolescent female underwent bone marrow transplantation for relapsed leukemia and developed acute and chronic graft-versus-host disease and idiopathic pneumonia syndrome. Her lung disease responded to large doses of methylprednisolone but evolved to pulmonary fibrosis and pneumomediastinum and subcutaneous emphysema in the convalescent period. Pulmonary function tests revealed a restrictive pattern. Pneumomediastinum and subcutaneous emphysema are complications not only of obstructive but also of restrictive lung disease and vary with respect to time of onset. PMID- 10689723 TI - Nutritional strategies in cardiovascular disease control: an update on vitamins and conditionally essential nutrients. AB - Several nutritional interventions for cardiovascular disease (CVD) prevention and therapy have recently appeared in the biomedical literature. These include appropriate use of several vitamins (E, C, B6, folate) and conditionally essential nutrients (CoQ10, L-arginine, propionyl L-carnitine). Possible undesirable consequences of long term nutritional supplementation with vitamin E and of adverse drug-nutrient interactions between the statins and CoQ10 are also considered. Although additional intervention studies are needed, current scientific evidence generally supports nutritional supplementation with these nutrients as an effective adjunctive strategy for CVD control. PMID- 10689724 TI - Reliability and validity of the Food Pyramid Self Efficacy Scale: use in coronary artery bypass patients. AB - Inappropriate dietary intake is associated with 5 of the 10 leading causes of U.S. death; coronary artery disease (CAD) ranks highest regardless of gender in people over the age of 65. Of the modifiable risk factors for CAD, two of four pertain to food choices. Although lifestyle habits can enhance or impair health, people's beliefs that they can motivate and regulate their own behavior (self efficacy) plays a crucial role in whether they even consider changing detrimental health habits. The Food Pyramid Self Efficacy Scale (FPSES) is an instrument to measure an elder's confidence in his/her ability to choose healthy food items in a variety of situations. The purpose of this study was to determine the reliability and validity of the FPSES. Thirty postoperative CABG patients participated (mean age 70.4). FPSES test-retest (r = 0.78, p = 0.008); coefficient alpha = 0.92. Six content experts judged the FPSES (content validity index [CVI] = 0.85). Construct validity of the instrument was achieved through hypothesis testing, supporting the statement that the higher the nutritional risk, the greater the functional decline (r = 0.37, p = 0.05). As many of the health problems associated with the elderly are preventable or controllable through health promotion, it is vital that measures exist to determine a person's confidence that one believes in the capability to change to healthy eating behaviors. PMID- 10689725 TI - Suppose a Perclose. AB - The development of coronary interventional strategies and devices has provided an alternative to surgery for coronary artery disease. The femoral artery is the most common access site for therapeutic cardiac interventions. The methods to achieve hemostasis post procedure vary from conventional, where the arterial puncture site is externally compressed, to novel, where the femoral artery is sealed with a device. One such device is a suture mediated percutaneous vascular closure system developed by Perclose, Incorporated. It provides a safe, effective method to achieve immediate hemostasis of the femoral artery. The vascular complication rate is only 0.78%, time to hemostasis is within 1.3 minutes, and time to ambulation in within 5.5 minutes. These advantages provide improved patient comfort and have a positive economic impact. PMID- 10689726 TI - Collaboration, problem solving, reevaluation: foundation for the Heart Center of Excellence. AB - Today's rapidly changing health care environment has thrust upon the caregivers, patients, and their families many challenges. A multidisciplinary team continuously analyzes strengths, weaknesses, and threats to the success of heart centers. Prolonged hospitalization for cardiac surgery patients is consistently identified as a priority with multiple opportunities for improvement. All patients are monitored for variables known to impact length of stay including same day admission, intubation time, out of bed interval following extubation, and delay in postoperative transfer to the progressive cardiac care unit (PCCU). Utilizing collaboration, problem solving, and reevaluation, known as CPR techniques, postoperative intubation time, and the out of bed interval following extubation, have both dramatically decreased. Despite declines in elective, same day admissions and fewer first day postoperative transfers, length of stay following cardiac surgery has also declined. The continuum of patient care has been established from preadmission to postdischarge and up to one year following cardiac surgery. Utilizing CPR techniques allowed us to achieve a best practice model for successfully improving outcomes in heart centers. PMID- 10689727 TI - Effective treatment of severe hypertension. PMID- 10689728 TI - New guidelines for the management of hypertension: latest perspective on an old problem. PMID- 10689729 TI - The Working Group on Cardiovascular Nursing: a European forum for international collaboration in cardiac care. PMID- 10689730 TI - What is the most common arrhythmia following cardiac revascularization? PMID- 10689731 TI - Is aggressive lipid lowering as effective as angioplasty in some cardiac patients? PMID- 10689732 TI - Differentiation and proliferation of pulmonary neuroendocrine cells. AB - In this review article the morphological profiles of pulmonary neuroendocrine cells (PNEC) in experimental animals and humans are described. Although the mechanisms of differentiation and proliferation of neuroendocrine cells in the airway epithelium remain to be solved, several experimental studies using explant culture and cell culture systems of fetal animal lungs have been performed to clarify fundamental phenomena associated with neuroendocrine differentiation and proliferation. Experimental animal studies using chronic hypoxia, toxic substances and carcinogens have succeeded in inducing alterations in PNEC systems, and these studies have elucidated the reactions of PNEC in cell injury and inflammation, and functional aspects of PNEC in disease conditions. Human pulmonary neuroendocrine tumors include various histological subtypes, and show divergent morphological and biological varieties. Molecular abnormalities of small cell carcinoma, the most aggressive subtype of pulmonary neuroendocrine tumors, have been extensively studied, but the mechanism of neuroendocrine differentiation of this tumor is still largely unknown. PNEC share common phenotypes with neuronal cells, and developmental studies have begun contributed evidence that similar transcriptional networks, including active and repressive basic helix-loop-helix (bHLH) factors, function in the differentiation of both PNEC and neuronal cells. Such a bHLH network may also play a central role in determining cell differentiation in lung carcinomas. Further studies of the neuronal bHLH network, its regulatory system and related signal transduction pathways, will be required for understanding the mechanisms of neuroendocrine differentiation and proliferation in normal and pathological lung conditions. PMID- 10689733 TI - [The Wiener klinishce Wochenschrift at the beginning of the 3rd century]. PMID- 10689734 TI - [Pathophysiology of acute renal failure at the cellular level]. AB - The influence of inflammation on post-ischemic acute renal failure (ARF) has only recently be appreciated. In this review we therefore discuss the cellular events occurring in ARF with special emphasis on the impact of inflammatory processes on the pathogenesis of ARF. Furthermore, the spectrum of injury leading to sublethal or lethal cell damage and the time course, occurrence and regulation of the two distinct forms of cell death, necrosis and apoptosis will be described extensively. Especially apoptosis and its regulation has been studied only marginally in the setting of ischemic ARF. This overview is mainly focused on tubular cell injury since tubular epithelial cells are the major victims of ischemia whereas cells inside the glomerular tuft show only little pathology. The models of tubular injury described in this paper are ranging from primary cultures of isolated human tubular epithelial cells to experimental ischemic renal failure in rats, and to clinical settings of human ischemic ARF. The cellular events highlighted in this review are the influence of the expression of cellular adhesion molecules on the pathophysiology of ARF, and the regulation and time course of apoptosis. Examples of these processes are being illustrated by figures exhibiting morphology and immunohistochemistry of cell proliferation and cell death regulatory proteins. PMID- 10689735 TI - Normalization of orbital arterial blood flow in non-ischemic central retinal vein occlusion after 6 months. AB - INTRODUCTION: Central retinal vein occlusion (CRVO) is a common vascular disorder and may lead to blindness. The aim of the study was to obtain information about the possible imbalance and recovery of orbital arterial blood flow in non ischemic CRVO. METHODS: Vascular resistance (pulsatility index-PI) in the orbital arteries of 14 patients with non-ischemic CRVO was examined within 3 weeks after onset of CRVO and 6 months later. The control group consisted of 14 age- and sex matched healthy control eyes. RESULTS: PI was increased in all orbital arteries of CRVO eyes measured within 3 weeks after the onset. Normal PI values were recorded in the same retrobulbar arteries, and re-measured 6 months later. DISCUSSION: There is an increase in vascular resistance in all orbital arteries at the onset of non-ischemic CRVO, followed by a recovery of vascular resistance to normal levels 6 months later. The increase at the onset may be caused by arterial vasospasm or by intraocular hemostasis affecting the afferent arteries. PMID- 10689736 TI - Histochemical changes in the rectal mucosa of diabetic patients with and without diarrhea or constipation. AB - Sixty-four diabetic patients, 35 with diarrhea, 15 with constipation and 14 without stool problems, and forty healthy subjects, were subjected to rectosigmoidoscopy. During rectosigmoidoscopy, rectal biopsy specimens for histological and histochemical analysis were obtained. Histological findings of nonspecific colitis in 25 out of 64 diabetic patients were uniformly distributed among the three groups (p = 0.959). However, the finding was slightly more common in diabetic patients than in controls (eight out of 40 control subjects, p = 0.043). A positive PAS reaction was observed in 30 out of 64 diabetic patients and was also uniformly distributed among the three groups (p = 0.508), but was significantly more common among diabetic patients than controls (three out of 40, p < 0.001). A positive reaction to cholesterol was found in 46 out of 64 diabetic patients, also uniformly distributed among the three groups (p = 0.773). It was significantly more common in diabetic patients than in controls (nine out of 40, p < 0.001). Reactions of the rectal mucosa histological specimens to glycogen and triglycerides were negative, both in diabetic patients and in controls. In conclusion, it appears that stool problems among our diabetic patients were not related to the positivity of PAS or to the positive cholesterol reaction in the rectal mucosa histological specimens. Since positive findings of both reactions were more common in specimens taken from diabetic patients than in controls, positive reactions might be related to metabolic disturbances in diabetic patients. PMID- 10689737 TI - [Pericardial effusion in celiac disease--an incidental finding?]. AB - OBJECTIVE: Ultrasound revealed evidence of pericardial effusion in 13 out of 26 children with coeliac disease. In a prospective study, we tried to analyse the causes underlying this high incidence of pericardial effusion. PATIENTS AND METHODS: Twenty-six patients were evaluated. Coeliac disease was diagnosed by intestinal biopsy. All children underwent sonography and a laboratory work-up including endomysial antibodies and serum selenium and iron concentrations. RESULTS: Patients with pericardial fluid showed no difference compared to those without effusion in regard to ECG, chest x-ray, red and white blood cell count, serum enzymes, serum protein as well as iron levels. The mean value of serum selenium was lower and endomysial antibody titre was higher in patients with pericardial effusion. However, due to the wide range, a clear distinction between the two groups was impossible. In all other investigated parameters there was no difference between patients with and without pericardial effusion. Patients with effusion had a higher frequency of viral infection. CONCLUSION: The high incidence of pericardial effusion in patients with coeliac disease appears to be governed by a multifactorial mechanism. A high endomysial antibody titre as well as selenium deficiency may play a role as a predisposing factor. Viral infection due to reduced immunological competence in conjunction with a hampered ability to eliminate toxic free radicals might cause blood vessel dysfunction, resulting in (asymptomatic) pericardial effusion. The fact that most of these patients were diagnosed during the cold season, with anamnestic evidence of viral infection shortly before the diagnosis, and the fact that adult patients with dilative cardiomypathy show a greater prevalence of coeliac disease, supports the view that coeliac disease is systemic in nature. PMID- 10689738 TI - Pancreatic elastase 1 in stool: variations within one stool passage and individual changes from day to day. AB - Concentration of fecal pancreatic elastase 1 has been claimed to be a highly sensitive and specific noninvasive test for exocrine pancreatic function. The aim of our study was to investigate variations in elastase concentration within one stool passage and from day to day. For the analysis of the variation of fecal elastase within one stool passage, we utilized 3 different samples collected from 8 patients. Further, we assessed the individual day to day variation of fecal elastase using stools collected on 3 consecutive days from 40 patients. For the determination of pancreatic elastase 1 in stool we used an ELISA kit. We found a relatively considerable variation of fecal elastase concentration within one stool passage (n = 8, mean CV = 22%, range 4.6-83.1%) and from day to day (n = 40; mean CV = 26%, range 2.4-61.1%). Therefore, we recommend routine analysis of more than 1 stool sample collected on different days for fecal elastase and to use a borderline area of +/- 25% of the recommended cut off of 200 micrograms/g stool for the diagnosis of pancreatic insufficiency. PMID- 10689739 TI - [Josef Weinlechner (1829-1906). Pioneer in Vienna pediatric surgery]. AB - The history of paediatric surgery in Vienna has not been clearly documented so far, especially its development during the second half of the nineteenth century. Josef Weinlechner (1829-1906) is one of the outstanding paediatric surgeons in Vienna during this time. We present an overview of his biography, his position as head of the department of surgery at the St. Anna Children's Hospital, and also discuss his publications concerning various aspects of paediatric surgery. Of particular interest was the discovery of his application for the qualification of a postdoctoral lecturer ("Habilitation") in the archives of the University of Vienna. In this application, Weinlechner refers to his specialisation as that of a paediatric surgeon. PMID- 10689740 TI - [Austrian Society of Lung Diseases and Tuberculosis: Consensus on Management of Chronic Obstructive Lung Diseases (COPD). 1999 Revision]. PMID- 10689741 TI - Sponsored congress attendance--does it happen, does it matter? PMID- 10689742 TI - Complimentary journeys to the World Congress of Gastroenterology--an inquiry of potential sponsors and beneficiaries. AB - One of the most effective tools of pharmaceutical marketing is the distribution of gifts to physicians whose magnitude remains ill defined. This anonymous survey determines the frequency with which physicians receive travel awards from drug companies to attend International Medical Conventions and attempts to obtain the recipients' opinion on ethical and legal issues related to such sponsorships. METHODS: A questionnaire was mailed to all members of the German Gastroenterological Association who had attended the most recent World Congress of Gastroenterology and to 30 pharmaceutical companies. Questions concerned the physician's role at the congress, the mode of payment for travel, lodging and convention fees as well as the attendees' opinion on ethical and legal issues related to sponsoring by pharmaceutical companies. RESULTS: 78% of the contacted physicians returned the questionnaire. 67% (95% CI [55, 80]) of them received compensation for their travel expenses by industry, and the majority of them stated that they would not have attended the congress if such sponsoring had not occurred. More than two thirds believed that sponsoring by drug companies neither interferes with ethical and legal issues nor affects prescribing behavior. Such opinions were more frequently expressed by sponsored than nonsponsored attendees (p = 0.003). 20% of the contacted drug companies returned the questionnaire, one of whom expressed concerns regarding the ethics of sponsorships. CONCLUSIONS: International conventions would suffer from a significant deprivation of attendance if the attendees' expenses were not subsidized by industry. Recognition of ethical and legal issues related to such sponsoring appears to be limited and requires further discussion within the medical community. PMID- 10689743 TI - Accuracy of 13C-urea breath test in clinical use for diagnosis of Helicobacter pylori infection. AB - The 13C-urea breath test (UBT) is a noninvasive test for diagnosis of Helicobacter pylori infection of gastric mucosa. The aim of this prospective study was to assess the accuracy of a simple UBT in clinical routine use. METHODS: The study population comprised of 100 patients (49 f, 51 m) requiring diagnostic upper GI endoscopy. One biopsy specimen was taken from the gastric antrum, body and fundus, respectively, for standard histological examination and one additional specimen from each location was transformed into transport medium for cultivation of H. pylori. After vaccination of the culture plates the biopsies were tested for urease activity (UAT). After recovery from endoscopy the patients had to pass an one liter endexspiratory breath sample before and 15 min after drinking 200 ml orange juice, pH 3.6, containing 75 mg of 13C-urea. 13CO2 was measured in the breath samples using isotope-selective nondispersive infrared spectrometry. RESULTS: Defining gold standard groups with all biopsy tests (from antrum and corpus) positive or negative the 13CO2 delta over baseline (DOB) cut off level of UBT was set at 6.5/1000 in order to best discriminate positive from negative patients (ROC analysis). UBT was positive in 37% of all subjects. Taken UAT and histological examination together (positive when both tests were positive) UBT displayed a sensitivity of 92%, a specificity of 94%, a positive predictive value of 89%, and a negative predictive value of 94%. When including the results of culture sensitivity and negative predictive value reached almost 100%. The mean of the 13CO2-DOB values from H. pylori-positive duodenal or gastric ulcer patients did not differ from controls (H. pylori-positive patients without lesions). The 13CO2-DOB values of the ulcer group were correlated significantly with the active inflammatory component of gastritis in antrum, corpus, and fundus. CONCLUSION: UBT with this setup detects H. pylori infection in clinical routine use with high accuracy. The increase of exhaled 13CO2 does not predict ulcer disease but reflects the degree of active inflammation of gastric mucosa. PMID- 10689744 TI - [Fluorescence endoscopy in gastroenterology]. AB - Fluorescence endoscopy is a new technique which allows a better endoscopic detection of nonvisible or difficult detectable malignant or premalignant lesions. Exogenously applied sensitizers accumulate selectively in malignant lesions and induce fluorescence after illumination with light of adequate wavelength. However, also endogenous fluorophores, different located in malignant or benign lesions, induce a different autofluorescence in these tissues. Meanwhile fluorescence endoscopy is a widely spread technique in urology using 5 aminolevulinic acid sensitization. In gastroenterology this technique seems promising in the detection of early cancers or dysplasia in patients with Barrett's esophagus or ulcerative colitis. This paper describes the current status and future development of fluorescence endoscopy in gastroenterology. PMID- 10689745 TI - [The liver and hereditary hemorrhagic telangiectasia]. AB - Hereditary hemorrhagic teleangiectasia, or Rendu-Osler-Weber syndrome, is an autosomal dominant inherited disease characterized by vascular derangement in many organs. The vascular derangement includes teleangiectases, arteriovenous fistulas and aneurysms. Liver involvement in hereditary hemorrhagic teleangiectasia is a rare and sometimes severe disease which was unknown and mostly detected at autopsy until a few decades ago. Typical findings are vascular malformations and connective tissue formation with fibrosis and atypical cirrhosis. In the last years we observed five Osler patients with exclusive or prevailing involvement of the liver. An unambiguous diagnosis can be ascertained by means of a hazardous liver puncture with typical histological findings. Angiography allows a reliable identification of even minor vascular deformities. The present study was undertaken to demonstrate the courses of disease, the techniques of examination and the therapeutical options of this rare manifestation of Osler-disease. In every case one could observe hepatic malformations which were established as typical Osler findings with the assistance of histological and above all sonographical/color-Doppler sonographical devices. These special sonographic/color-Doppler-sonographic features make it possible to give up the histological diagnostics or exhaustive investigations by means of angiography, computer-tomography or MRI. PMID- 10689746 TI - Mechanic intestinal obstruction--a possible presentation of perforated appendicitis. AB - A 61-year-old man presented with diffuse abdominal pain, diarrhea, vomiting and fever. On the initial diagnosis of gastroenteritis the patient received the antibiotic ofloxacine for one week. On admission plain abdominal radiograph suggested a mechanic intestinal obstruction. In computed tomography a conglomerate tumor in the ileocecal region was seen and the patient underwent laparotomy. The conglomerate tumor was mobilized and an abscess opened, which was caused by a perforated appendicitis. After the operation the patient improved immediately and had an uneventful postoperative course. He was released and did not suffer from gastrointestinal symptoms the following 16 months of follow-up. The present case shall set forth that perforated appendicitis can clinically present as intestinal obstruction. Although a rare complication, perforated appendicitis should therefore even be considered in cases of mechanic intestinal obstruction of unknown cause. PMID- 10689747 TI - [Pneumopericardium and retroperitoneal and scrotal emphysema after endoscopic papillotomy]. AB - Retroduodenal perforation is considered a rare but serious complication of endoscopic papillotomy. We report on a patient suffering from a stenotic occlusion of the ductus hepatico-choledochus on whom a pre-cut via needle-knife was performed in order to place a stent. Hours after the procedure, the patient developed an extensive pneumoretroperitoneum, pneumopericardium and scrotal emphysema. The later have not been reported in relevant literature so far. PMID- 10689748 TI - [Colorectal carcinoma: prevention and early detection in an asymptomatic population--prevention in patients at risk--endoscopic diagnosis, therapy and after-care of polyps and carcinomas. German Society of Digestive and Metabolic Diseases/Study Group for Gastrointestinal Oncology]. PMID- 10689749 TI - [New developments in therapy of chronic hepatitis B. When are nucleoside analogs indicated?]. AB - Nucleoside analogues are promising agents for the treatment of chronic hepatitis B infection (HBV-DNA-positive by hybridization assay). The drug being studied most intensively is Lamivudine (Zeffix) which has recently been approved in Germany. When given orally once daily (100 mg) Lamivudine is well-tolerated and suppresses HBV-DNA to undetectable levels in the majority of patients. Since relapse is frequent when medication is stopped long-term treatment (at least until seroconversion of HBeAg) is warranted. Indications for lamivudine monotherapy are patients with chronic hepatitis B in which interferon (IFN) is contraindicated or patients who did not respond to a previous course of interferon. Further indications are the HBV-DNA-positive cirrhosis prior to liver transplantation (OLT) and the HBV-reinfection after OLT. The main problem of long term monotherapy with lamivudine is viral resistance. The clinical impact of the resistant mutants is often not clear. Withdrawal or even continuation of the medication may be acceptable approaches. Other nucleoside analogues like Entecavir or Adefovir are currently being tested in clinical studies. Famciclovir was investigated preferably in patients with decompensated liver disease or HBV reinfection after OLT. Because of conflicting results the drug should only be used under study conditions. In IFN-naive patients with chronic hepatitis B (and compensated liver disease) alpha-interferon is still the first-line therapy. With a standard course of interferon 30-40% of the patients become seronegative for HBeAg as compared with 16-17% when treated with lamivudine for twelve months. Combination of lamivudine and interferon is not more effective than IFN alone. In the future combined antiviral treatment is likely to replace monotherapy. PMID- 10689750 TI - [The MACH2 Study: value of inhibiting gastric acid secretion in eradication of Helicobacter pylori with a one week triple therapy protocol]. PMID- 10689751 TI - [The value of botulinum toxin injection in treatment of achalasia]. PMID- 10689752 TI - [High fiber diet or not, that is the question here--comments on prevention of colorectal carcinoma by dietary fiber]. PMID- 10689753 TI - [Comment on consensus report: Irritable bowel syndrome--definition, verifying the diagnosis, pathophysiology and therapeutic options]. PMID- 10689754 TI - Silica and lung cancer: hazard or risk. PMID- 10689755 TI - Epidemiological evidence on the carcinogenicity of silica: factors in scientific judgement. AB - In view of the extended debate and differing opinions on whether crystalline silica is a human carcinogen, we have reviewed a selection of epidemiological reports, to identify the areas of uncertainty and disagreement. We have chosen to examine the papers which in a recent review were considered to provide the least confounded examinations of an association between silica exposure and cancer risk. We also refer to a study of the mortality of coalminers very recently reported by ourselves and colleagues. We find that parts of the evidence are coherent but there are contradictions. On examination this resolves mostly into differences between types of studies. The three types of epidemiological study included are: (i) exposure-response studies, the most powerful for the confirmation of a relationship between a specific exposure and a health effect; (ii) descriptive studies in which incidence of disease in an exposed population is compared with that in a reference population; and (iii) studies of incidence of disease in subjects on silicosis case-registers. Descriptive studies frequently though not invariably suggest an excess lung cancer risk in silica exposed workers compared with the general population, but exposure-response studies consistently fail to confirm that the cause is exposure to quartz. A single exposure-response study of cristobalite suggests a positive relation. Both sets of evidence have weaknesses. There are uncertainties on whether the excess risks in the descriptive studies are related to silica exposure or to lifestyle, including smoking habits. There are doubts on whether the exposure estimates in some of the exposure-response studies were sufficiently reliable to detect a small risk or weak association, though they are unlikely to have missed a strong effect. Studies of subjects on silicosis case registers consistently show an excess of lung cancer, but it is not clear to what extent these increased risks represent a direct effect of silica exposure, a secondary effect of the silicosis, preferential inclusion of subjects suffering from the effects of smoking, or bias in diagnostic accuracy. This not unnaturally leads to differences in opinion, exacerbated by variations in the strength of proof required by different experts. The main scientific uncertainties in the evidence are: 1. Whether, in the descriptive studies, the excess lung cancer rates in silica-exposed workers are explicable in terms of smoking habits, socio-economic class differences and inappropriate comparison populations. Better smoking information and more carefully chosen comparison populations are needed; 2. Whether the exposure-response studies could have missed a real relationship between silica exposure and lung cancer, if one exists. Many of the exposure response studies were conducted with great care, but weaknesses, in the available data on which the exposure estimations were based, could have caused a real relationship of lung cancer and silica exposure to be missed. These studies were sufficiently powerful to demonstrate relationships of silica exposure with silicosis and silico-tuberculosis, so it is unlikely that they would have missed any but a small risk, or weak relationship, for lung cancer. Our own recent study of coalminers used uniquely detailed and reliable exposure data, and failed to demonstrate convincingly an increased risk. This negative finding, though, applies only to a dust in which the proportion of quartz in the dust is usually less than 10%. Exposure-response studies are needed, with high quality exposure estimates, in populations exposed to respirable dust of which crystalline silica comprises more than 10%; 3. Whether the excess cancer risks in subjects on silicosis registers are the result of selection and diagnostic bias. Given these difficulties, case-register studies may not be capable of giving a reliable answer to the central question, though they have been useful in pointing to the possibility of a can PMID- 10689756 TI - Inhalation study on exposure to bitumen fumes. Part 1: Development and validation of the equipment. AB - Bitumen fumes emitted during road paving or roofing contain polycyclic aromatic hydrocarbons (PAHs). Experimental studies have been previously performed to test the carcinogenic potency of bitumen fumes. Some of them have been criticised either on the grounds that the fume condensates were not representative of fumes to which humans are exposed or because the fumes were never characterised in terms of particle size and poorly in terms of composition and concentration in the chambers. For a nose-only inhalation study, we have evaluated the ability of a new fume generation system to deliver stable and reproducible atmospheres of bitumen fumes to an inhalation chamber and investigated the representativity of the fumes generated at a concentration level of 5 mg/m3. The fume generator comprises: (1) an insulated 20 l heated kettle (200 degrees C for bitumen); (2) an insulated inlet pipe with a needle valve to adjust the flow of the test compound from the kettle; (3) a fume generation chamber equipped with a series of interchangeable channels of different width. The fume concentration in the exposure chamber can be controlled by changing the channel width or by restricting the evaporation surface with aluminium foil, and/or by changing the flow rate. Samples of the atmosphere in the chamber were collected and analysed for quantitative determination of total particulate matter (TPM), soluble matter, benzo[a]pyrene (B[a]P) content of the fumes and other PAHs, and evaluation of the particle size distribution. The representativeness of the fumes has been tested by comparison with fumes generated in the Shell small-scale fume rig, which was previously validated against field fumes collected during paving operations. Evaporative losses from the filters during sampling, transport and storage have been also assessed. At 5 mg/m3 TPM, the agreement between laboratories was quite good for the TPM analyses and was good for the soluble matter and B[a]P. Evaporative losses may lead to underestimation of the true exposure level in the inhalation chambers but the use of an XAD-2 cartridge backup is one approach to partially recover losses which occur on the filter. The particle size distributions are somewhat different from those reported for fumes associated with roofing and indoor mastic laying works, in that we found more than 85% of particles to be smaller than 1 micron, compared with 40% particles in the previous analyses. In conclusion, this equipment allows reproducible generation of fumes at the 5 mg/m3 TPM that are fairly representative of those produced in the field with the same bitumen. PMID- 10689758 TI - Statistical modelling of the determinants of historical exposure to bitumen and polycyclic aromatic hydrocarbons among paving workers. AB - INTRODUCTION: An industrial hygiene database has been constructed for the exposure assessment in a study of cancer risk among asphalt workers. AIM: To create models of bitumen and polycyclic aromatic hydrocarbons (PAH) exposure intensity among paving workers. METHODS: Individual exposure measurements from pavers (N = 1581) were collected from 8 countries. Correlation patterns between exposure measures were examined and factors affecting exposure were identified using statistical modelling. RESULTS: Inhalable dust appeared to be a good proxy of bitumen fume exposure. Bitumen fume and vapour levels were not correlated. Benzo(a)pyrene level appeared to be a good indicator of PAH exposure. All exposures steadily declined over the last 20 years. Mastic laying, re-paving, surface dressing, oil gravel paving and asphalt temperature were significant determinants of bitumen exposure. Coal tar use dictated PAH exposure levels. DISCUSSION: Bitumen fume, vapour and PAH have different determinants of exposure. For paving workers, exposure intensity can be assessed on the basis of time period and production characteristics. PMID- 10689757 TI - Inhalation study on exposure to bitumen fumes. Part 2: Analytical results at two exposure levels. AB - During the hot application of bitumen-containing materials, e.g. in road paving or roofing, fumes are emitted that contain traces of polycyclic aromatic compounds (PACs). Although worker's exposure to these fumes is low, it might lead to health problems. For studying DNA adduct formation as a consequence of inhalation of bitumen fumes we developed and validated an inhalation system (a dynamic fume generator plus a nose only inhalation chamber). This paper presents and discusses the analytical results from the different laboratories involved in this study on the fumes sampled in the inhalation chamber during three series of experiments where the animals were exposed to fumes at the 5 mg/m3 and 50 mg/m3 level, coming from bitumen heated at 200 degrees C and, as a positive control, fumes from coal tar, heated to 110 degrees C at the 5 mg/m3 level. The following parameters were controlled: temperatures at different key places in the generator; humidity of the chamber; the bitumen or coal tar flow rate; and Total Particulate Matter (TPM). Analyses were performed for Benzene Soluble Matter (BSM), the EPA polycyclic aromatic hydrocarbon (PAH) mixture and for a number of heteroatom-containing PACs. The data show that the coal tar fumes produced at 110 degrees C were very volatile and that most of the differences in particulate matter found between the laboratories can be attributed to evaporative losses. The bitumen fumes boil 25-50 degrees C higher and contain higher boiling compounds. A comparison is made between the PAC exposure profiles for bitumen experiments aimed at 5 and 50 mg/m3. Although the same molecules are found in both fumes their proportion is dramatically different. This effect is largest with the 2- and 3-ring PACs, the ratio of the concentrations found in the 50 mg/m3 TPM concentration to that in the 5 mg/m3 experiment gradually declines from 5500 for acenaphthene to 500 for pyrene, for the 5-ring PACs this ratio is 20-30. As function of their vapour pressure, the ratios of the concentrations of the hetero PACs follow the same trend as that of the 16 EPA PAHs and are of the same order of magnitude. In conclusion, for the compounds investigated, the equipment delivers a fume atmosphere in a reproducible manner. The 50 mg/m3 bitumen fumes are not representatives of field fumes. The reason for these quantitative differences is unclear and further work would be needed to clarify this. Nevertheless it was felt that these fumes at 50 mg/m3 might be a useful tool for qualitative detection of DNA adducts in an animal exposure study. PMID- 10689759 TI - Designing an international industrial hygiene database of exposures among workers in the asphalt industry. AB - OBJECTIVES: The objective of this project was to construct a database of exposure measurements which would be used to retrospectively assess the intensity of various exposures in an epidemiological study of cancer risk among asphalt workers. METHODS: The database was developed as a stand-alone Microsoft Access 2.0 application, which could work in each of the national centres. Exposure data included in the database comprised measurements of exposure levels, plus supplementary information on production characteristics which was analogous to that used to describe companies enrolled in the study. RESULTS AND DISCUSSION: The database has been successfully implemented in eight countries, demonstrating the flexibility and data security features adequate to the task. The database allowed retrieval and consistent coding of 38 data sets of which 34 have never been described in peer-reviewed scientific literature. We were able to collect most of the data intended. As of February 1999 the database consisted of 2007 sets of measurements from persons or locations. The measurements appeared to be free from any obvious bias. CONCLUSIONS: The methodology embodied in the creation of the database can be usefully employed to develop exposure assessment tools in epidemiological studies. PMID- 10689761 TI - Confusion about the precision of asbestos fibres counting by electron microscopy. PMID- 10689760 TI - In vitro and in vivo tests for determination of the pathogenicity of quartz, diatomaceous earth, mordenite and clinoptilolite. AB - The effects of samples of crystalline quartz, diatomaceous earth, mordenite and clinoptilolite were investigated in vitro (as concerns erythrocyte haemolysis and lactate dehydrogenase (LDH) release from peritoneal macrophages) and in vivo (on LDH, protein and phospholipids in rat bronchoalveolar lavage (BAL), and phospholipids in rat lung tissue). The respirable mineral samples were instilled intratracheally. Determinations in the BAL were carried out after 15, 60 and 180 days, and in the lung tissue after 90, 180 and 360 days. Quartz DQ and quartz FQ induced acute, subacute and chronic inflammation and progressive fibrosis. However, due to the Al2O3 contamination on the surface of the particles quartz FQ caused a delayed response in vivo. Diatomaceous earth produced acute/subacute inflammation that gradually became more moderate after 60 days. Clinoptilolite was inert, whereas the other zeolite sample, mordenite, was cytotoxic in vivo. The reason for this was presumably the needle and rod-shaped particles in the mordenite samples. The investigation revealed that different in vitro and in vivo methods canprovide valuable data concerning the pulmonary toxicity of minerals. PMID- 10689762 TI - An unusual case of self-treatment. PMID- 10689763 TI - Lack of respect for practitioners alarming. PMID- 10689764 TI - Calculating the impact of clinical governance. PMID- 10689765 TI - Tea breaks could help calm fluoride debate. PMID- 10689766 TI - Retention of permanent incisors by mesiodens: a family affair. AB - The term mesiodens refers to a supernumerary tooth that is present in the midline of the maxilla between the two central incisors. One or two mesiodentes may be present. We present a rare case of two sisters, in both of whom a pair of mesiodentes caused the retention of permanent incisors. They were referred to our hospital with asymptomatic delayed eruption of upper incisors. This article is written to point out genetic factors as the possible origin of supernumerary teeth. PMID- 10689767 TI - Videoconferencing: what are the benefits for dental practice? AB - For more than 3 years members of the TeleDent team from Bristol University have been looking at the potential of videoconferencing technology for dentistry. Here they explain what videoconferencing is and how it can help the GDP. They discuss examples of its most promising uses for the profession, which include professional updating and providing diagnostic support at a distance. They describe the equipment that is needed, the different types of system available and give an indication of costs. A suggested procedure for using the technology for remote referrals is outlined. 'Store and forward' techniques are also discussed. These do not involve a live video but involve the sending of static electronic files. This approach is compared with videoconferencing, and the article looks at the question of which will be best suited to the GDP, and for what purposes. PMID- 10689768 TI - Delayed and immediate hypersensitivity reactions associated with the use of amalgam. AB - Hypersensitivity to the constituents of dental amalgam is uncommon. When it occurs it typically manifests itself as a lichenoid reaction involving a delayed, type IV, cell-mediated hypersensitivity response. Rarely, a more acute and generalised response can occur involving both the oral mucosa and skin. We describe two cases that illustrate the presentation and management of these two types of reaction. PMID- 10689769 TI - Dental care for the patient with a cleft lip and palate. Part 1: From birth to the mixed dentition stage. AB - This is the first of two articles looking at dental care for the patient with a cleft lip and palate. Part 1 looks at the needs of the child with a cleft lip and palate from birth through to the mixed dentition stage. PMID- 10689770 TI - A study of the career development of male and female dental practitioners. AB - AIM: The aim of the study was to determine differences between male and female dental practitioners in the positions they occupy within their employment, and to analyse the correlates of such differences. METHOD: Postal questionnaire survey of a 1 in 10 sample of individuals taken from the General Dental Council register. RESULTS: Female dental practitioners occupy lower positions in the employment hierarchies of the Community Dental Service and the Hospital Dental Service. Women general dental practitioners are significantly less likely to be sole proprietor of, or a partner in, a general practice. Ownership of a general practice is related to: sex, age, number of years qualified, number of children, and hours worked. Consultants in the Hospital Dental Service are more likely to be male, older, to have been qualified longer and (obviously) to hold more additional qualifications than their non-consultant colleagues. Senior Dental Officers and Directors of the Community Dental Service are more likely to be male, work longer hours and (again obviously) to hold more additional qualifications than Community Dental Officers. CONCLUSIONS: Differences exist between male and female dental practitioners in the positions they occupy within employment hierarchies. Age, length of time since qualification and the acquisition of additional qualifications are consistently found to differentiate dental practitioners' status. PMID- 10689771 TI - Intensity of bacteraemia associated with conservative dental procedures in children. AB - OBJECTIVES: To explore the individual dento-gingival manipulative procedures that together lead to the placement of a restoration and to estimate the associated intensity of bacteraemia. PATIENTS AND METHODS: Healthy children receiving dental treatment under general anaesthesia provided blood samples 30 seconds after one of four dento-gingival manipulative procedures: 1. Placement of rubber dam, 2. Use of the high speed drill, 3. Use of the slow speed drill, and 4. Placement of matrix band and wedge. Blood cultures were processed to give the percentage prevalence of bacteraemia, the intensity of organisms per millilitre of blood and the identity of the organisms cultured. RESULTS: A total of 257 children were recruited to the study. The percentage positive prevalence of blood cultures was baseline--9.3%, rubber dam placement--31.4%, slow drill--12.2%, fast drill--4.3%, and matrix band and wedge--32.1%. The intensity of bacteraemia was baseline--1.2 cfu, rubber dam placement--1,962 cfu, slow drill--0.3 cfu, fast drill--1.9 cfu, matrix band and wedge--4.8 cfu. CONCLUSIONS: These data indicate that dento gingival manipulative procedures comprising a simple dental restoration can lead to a bacteraemia comparable to that from dental extractions. It is suggested that these data may indicate the need for antibiotic prophylaxis for some aspects of conservative dentistry. PMID- 10689772 TI - Perceptions of general dental practitioners of a local secondary care service in restorative dentistry. AB - AIM: A major role of the hospital based secondary care service in restorative dentistry is to accept referrals in order to formulate treatment plans. The aim of this survey was to improve service provision at Newcastle Dental Hospital by establishing baseline quality perceptions from referring general dental practitioners (GDPs). METHOD: A postal questionnaire was sent to 393 randomly selected local GDPs, with a response rate of 67%. RESULTS: Results indicated great demand for advice and treatment which was higher than expected for temporomandibular joint problems. 42% of GDPs felt that treatment plans were not helpful and the period of waiting for a response following consultation too long. The reasons for this perception are discussed. CONCLUSIONS: The need for better communication between primary and secondary care was highlighted, as was the distinct preference of GDPs for hospital consultants rather than registered specialist practitioners to carry out specialist treatment. PMID- 10689773 TI - Me gums made me do it. PMID- 10689774 TI - Surveillance of surgical site infections. PMID- 10689775 TI - Enhanced surveillance of neonatal group B streptococcal disease. PMID- 10689776 TI - Mechanism of action of CpG DNA. PMID- 10689777 TI - Oligodeoxyribonucleotides with 5'-ACGT-3' or 5'-TCGA-3' sequence induce production of interferons. PMID- 10689778 TI - Macrophage activation by immunostimulatory DNA. AB - Macrophage/dendritic cells and B cells remain the only cell types where direct responses to CpG DNA are well established. The role of macrophages in vivo in DNA clearance and the potent cytokine induction in macrophages and dendritic cells places them in the central role in the in vivo response to foreign DNA. Although responses to DNA are unlikely to evolve and be retained if they are not significant in the immune response to infection, the relative contributions of DNA and other stimulators of the innate immune recognition of foreign organisms is difficult to assess. Although CpG DNA and LPS have similar actions, significant differences are emerging that make the use of DNA as a therapeutic immunostimulatory molecule feasible. The macrophage response to DNA generates cytokines favouring the development of Th1-type immunity, and active oligonucleotides now show promise as Th1-promoting adjuvants and as allergy treatments. PMID- 10689779 TI - Consequences of bacterial CpG DNA-driven activation of antigen-presenting cells. PMID- 10689780 TI - Signal transduction pathways activated by CpG-DNA. AB - While more and more attention has been paid to CpG-DNA with respect to its usefulness as an adjuvant, its molecular mechanism of action is less well defined. Over the last few years, at least two major signalling pathways have been shown to be utilized by CpG-DNA: the NF-kappa B activation pathway and the stress-kinase pathway. Direct downstream events of these pathways are induction of transcriptional activity of NF-kappa B and transcriptional activity of AP-1. As far as investigated, CpG-DNA uses signal transduction pathways originally described for other stimuli, such as LPS, IL-1 or TNF. Therefore, to us, the prime question is: where does CpG-DNA-induced signalling enter these known pathways? This raises questions about the existence of a CpG-DNA-sequence specific receptor. Several points of evidence support the probability of the existence of a cellular receptor: There is a strong motif (unmethylated CpG) dependency for CpG-DNA-induced signalling. There is cell-type specificity. Dendritic cells, macrophages and B cells respond to CpG-DNA, but other cell types, such as fibroblasts and T cells, do not. In addition, classic signal transduction pathways are rapidly switched on in a parallel manner, as is known for other receptors. Using competing non-CpG ODNs and inhibitors of endosomal acidification, some evidence has been obtained that CpG ODNs are taken up into endosomes by a CpG-independent receptor, followed by a pH-dependent step before signalling starts. A model based on these findings is proposed in Fig. 4. Nevertheless, other receptor-independent activities of CpG-DNA cannot yet be ruled out. Although unlikely, we should consider the possibility that CpG-DNA directly interacts with cellular nucleic acids either by direct hybridization with complementary nucleotides or by formation of DNA triplexes (VASQUEZ and WILSON 1998). While these possibilities have been explored by antisense technology, using a huge variety of ODNs, there is no experimental evidence that such interactions are important for the activity of CpG-DNA. In this context, it is important to note that DNA, especially phosphothioate-stabilized ODNs with poly-G stretches, have substantial CpG-independent activities, although these activities seem not to depend on specific, antisense-like DNA-DNA interactions (PISETSKY 1996). One good example comes from experiments using ODNs on primary T cells. Co-stimulation of CD3-primed T cells with CpG ODN leads to a significant increase of IL-2 secretion and proliferation; however, these effects are CpG independent (K. Heeg, personal communication). Remarkably, these poly-G stretches seem to be inactive when transferred to double-stranded DNAs, such as plasmid DNA (WLOCH et al. 1998). In contrast, to my knowledge, no immune-stimulatory effect of bacterial DNA has been described that can not be abolished by CpG-specific methylation. Taken together, CpG-dependent and CpG-independent activities must be distinguished from one another. Among these effects, CpG-dependent signalling is better defined. Much effort is going into the investigation of the pharmacological applications of CpG-DNA. Once CpG-receptor-like structures are known, the question of the physiological role of CpG-DNA can be tackled. PMID- 10689781 TI - CpG DNA co-stimulates antigen-reactive T cells. PMID- 10689782 TI - Role of type I interferons in T cell activation induced by CpG DNA. PMID- 10689783 TI - Hematopoietic remodeling triggered by CpG DNA. PMID- 10689784 TI - CpG DNA augments the immunogenicity of plasmid DNA vaccines. PMID- 10689785 TI - The role of bacterial DNA in autoantibody induction. AB - Bacterial DNA has potent immunological properties that can stimulate the immune system in SLE in both specific and non-specific ways. As such, this molecule may play an important role in disease pathogenesis, because it can exert immunomodulatory activity and function as a molecular mimic. Future studies will hopefully both determine the role of foreign nucleic acids in the induction of autoantibodies and lead to strategies for their elimination. PMID- 10689786 TI - CpG DNA in cancer immunotherapy. PMID- 10689787 TI - Use of CpG DNA for enhancing specific immune responses. AB - CpG ODN, owing to its wide range of immunostimulatory effects has been found to be a potent Th1-type adjuvant that is effective with virtually any type of antigen, although responses are less impressive with PS than protein antigens. The use of CpG ODN as an adjuvant may allow the development of vaccines against a wider range of diseases, which could include therapeutic vaccines for chronic infections or cancer, effective pediatric vaccines for newborns, and easily delivered mucosal vaccines. PMID- 10689788 TI - Immunostimulatory-sequence DNA is an effective mucosal adjuvant. PMID- 10689789 TI - CpG DNA switches on Th1 immunity and modulates antigen-presenting cell function. PMID- 10689790 TI - Effects of CpG DNA on Th1/Th2 balance in asthma. AB - Thus, in our studies, we demonstrated that CpG ODN are effective in preventing the development of eosinophilic airway inflammation and bronchial hyper reactivity in a murine model of asthma. Antigen-associated elevation of serum IgE levels is also suppressed. CpG ODN, administered in conjunction with antigen, is also effective in down-regulation of established Th2 responses. This protection is neither murine strain-dependent nor model-dependent. Although these effects of CpG ODN are associated with the induction of the Th1 cytokines IFN-gamma and IL 12, neither cytokine is absolutely required for the protection. These results suggest that CpG ODN may be an effective immunomodulatory agent in the treatment, and possibly prevention, of asthma. PMID- 10689791 TI - Responses of human B cells to DNA and phosphorothioate oligodeoxynucleotides. AB - Emerging information has documented that certain DNA and sODNs can be both immunogenic and immunostimulatory. sODNs, but not DNA, induce T-cell-independent polyclonal activation of human B cells by engaging cell-surface receptors. Manifestations of sODN-induced human B-cell activation include expression of activation markers, proliferation, Ig production and anti-DNA antibody production. IL-2 and intact T cells enhanced B-cell responses to sODNs but were not required. Monocytes also provided a modest enhancement of human B-cell responses induced by sODNs. The chemical nature of sODNs capable of stimulating human B cells and the specific cell-surface receptors involved have not been completely delineated. Further studies will be necessary to elucidate the potential role of stimulatory sODNs in disease pathogenesis and to develop a means to employ ODNs as therapeutic agents in humans. PMID- 10689792 TI - [Prevalence of Chlamydia infections in breeding sows and their importance in reproductive failure]. AB - To investigate the prevalence of chlamydial infection and their significance for reproductive disorders in sow breeding herds in Germany, blood samples of 1493 pigs were taken for a serological survey by enzyme-linked-immunosorbent-assay (ELISA). Antibodies to Chlamydiae were found in 33% of the samples, in all herds investigated responders could be detected. The rate of seropositive animals in different farms varied from 4.3% to 72.7%. The percentage of positive responders in the farms correlated positively with the occurrence of MMA-syndrome (mastitis, metritis, agalactia), return to oestrus and diseases of the piglets. Also these herds showed less weaned piglets per sow and litter. Comparison of seronegative and seropositive sows within single farms revealed also worse reproductive data for seropositive sows. A significant relationship could be found between farms with a high quota of sero-positive sows and poor hygiene status as well as poultry keeping. As a second step 124 cervical swabs and 9 aborted piglets were investigated for chlamydial antigen by capture-ELISA and polymerase chain reaction (PCR). Using the capture-ELISA for investigation only 3 probes with chlamydial antigen could be detected, however, examination by the more sensible PCR gave positive results in 50% of the probes. 20% of the PCR-positive animals were clinically healthy sows, 80% of the PCR-positive probes originated from sows with reproductive disorders. A significant relationship could be shown between PCR-positive probes and the incidence of abortion and litters with stillborn piglets and piglets with low viability. Swabs from 93 of the 124 sows were investigated as well for other bacterial pathogens of reproductive disorders. A high degree of micro-organisms of different species could be detected in 70% of the samples of sows with reproductive disorders and in 35% of the samples of clinically healthy sows. Species differentiation of the chlamydial antigen positive samples was done by southern blot hybridisation. Herewith C. psittaci could be diagnosed in all positive samples. Additionally 8 probes revealed a mixed infection with C. psittaci and C. trachomatis. The results of the present study show, that the prevalence of chlamydial infections in breeding herds is high and underline the importance of chlamydial infections for reproductive disorders. Single chlamydial infections as well as mixed infections with other pathogens must be considered. PMID- 10689793 TI - [Significance of respiratory compensation in acidosis in calves]. AB - The respiratory component PvCO2 of acid-base-status was observed in n = 36 calves (age: x +/- s = 8.7 +/- 5.0 d) with neonatal diarrhea and an acidosis (venous blood-pH: < 7.30; x +/- s = 7.08 +/- 0.15). In n = 10 (28%) calves with a severe metabolic acidosis (pH: x +/- s = 7.03 +/- 0.12; BE: x +/- s = -22.1 +/- 5.3 mmol/l) the PvCO2 was decreased < 5.3 kPa (x +/- s = 4.5 +/- 0.5 kPa) and showed a distinct respiratory compensation. A PvCO2 between 5.3-6.7 kPa (x +/- s = 6.0 +/- 0.4 kPa) was observed in n = 16 (44%) acidotic calves (pH: x +/- s = 7.11 +/- 0.13; BE: x +/- s = -15.2 +/- 7.4 mmol/l). These n = 26 (72%) calves showed a simple metabolic acidosis which is well known for calves with neonatal diarrhea. The remaining n = 10 (28%) calves showed an increase of the PvCO2 > 6.7 kPa (x +/ s = 8.0 +/- 1.5 kPa). These animals had a mixed respiratory-metabolic acidosis (pH: x +/- s = 7.08 +/- 0.20; BE: x +/- s = -13.9 +/- 10.3 mmol/l), as the decrease of the pH could not be determined by the decreased metabolic component HCO3- of acid-base-status alone. Calves which died during hospitalization and calves with a PvCO2 > 6.7 kPa tended to be younger and showed partially significant lower values for the parameters of oxygen-supply PvO2 and SvO2. Lactate was significantly higher in dying calves but not in calves with a mixed acidosis which on the other hand were more dehydrated. The functional capacity of respiratory compensation of acidotic disorders in the calves studied promised to be almost the same as in dog and man. One reason for the failure of respiratory compensation in some calves could be a more severe hypovolemia. With the use of "venous hypoxemia" (decrease PvO2 and decrease SvO2) the detection of tissue hypoxia was easier than with lactate concentration. PMID- 10689794 TI - [Possibilities of oral iron supplementation for maintaining health status in calves]. AB - Oral supply of 100 mg iron as bivalent cations per day effectively prevented clinical and subclinical symptoms of anaemia in calves. Additionally, male and female calves reached optimal growth values. The supplementation of iron to alleviate neonatal masked anaemia ("physiological anaemia") should start on the first day of life. This measure can be terminated at the end of the praeruminal stage of the development, as young calves are able to take up all ferric ions contained in the diet. The real extend of the iron deficiency of calves could not be determined by clinical symptoms. As a practical measure it is recommended to use the packed cell volume test. PMID- 10689795 TI - Tissue distribution and disposition kinetics of enrofloxacin in healthy and E. coli infected broilers. AB - Concentrations of enrofloxacin equivalent activity were determined by microbiological assay in the plasma of healthy and E. coli-infected broilers following single intravenous and oral administrations at 10 mg/kg. Tissue distribution and residue-depletion following multiple oral doses (10 mg/kg for 3 successive days) were investigated. Pharmacokinetic variables were determined using compartmental and non-compartmental analytical methods. Plasma enrofloxacin concentrations after intravenous dosing to healthy and infected birds were best described by a two-compartments model. Enrofloxacin concentrations in plasma of infected birds were lower than those of healthy ones. The disposition kinetics of intravenously administered drug in healthy and infected birds were somewhat different. The elimination half-life (t1/2 beta) was 4.75 vs. 3.63 h; mean residence time (MRT) was 6.72 vs 4.90 h; apparent volume of the central compartment (Vc) was 1.11 vs 1.57 l/kg; rate constant for transfer from peripheral to central compartment (k21) was 1.15 vs 1.41 h-1 and total body clearance (ClB) was 0.35 vs 0.53 l/h/kg in healthy and infected birds, respectively. After oral administration, the absorption half-life (t1/2abs) in the infected birds was significantly longer than in healthy birds, while elimination half-life (t1/2el) and MRT were significantly shorter. Bioavailability was higher in infected birds (72.50%) as compared to healthy ones (69.78%). Enrofloxacin was detected in the tissues of healthy and infected birds after daily oral dosing of 10 mg/kg for 3 days. It was more concentrated in liver, kidney, and breast muscle. The minimal inhibitory concentration (MIC) of enrofloxacin against E. coli was 0.064 microgram/ml. On the basis of maintaining enrofloxacin plasma concentrations over the MIC, a dose of 10 mg/kg given intravenously every 20.14 hrs or orally every 20.86 hrs should provide tissue concentrations effective against E. coli infection in chickens. PMID- 10689796 TI - Relationship between certain physical properties of cervical mucus and fertility in cows. AB - The object of the present investigation was to determine several physical properties of cervical mucus in spontaneous oestrus cows in relation to fertility. Because, it is very difficult to determine the exact time of the beginning of oestrus in cows, the aim of the present study was to investigate if physical properties of cervical mucus at the time of artificial insemination (AI) are related to conception or not. A total of 93 cows of Friesian breed were used. The animals exhibited spontaneous oestrus, without being submitted to any hormonal treatment. Samples of cervical mucus were collected 5-30 min before AI and pH, viscosity, spinnbarkeit (spinability), crystallization and penetration were measured. Pregnancy diagnosis was performed by rectal palpation 3 months later. The results obtained from cows that conceived (44 animals), compared to those obtained from cows that did not conceive (49 animals), were the following: a) viscosity had been significantly lower (p < 0.05), b) crystallization had been significantly higher (p < 0.05), and c) pH, spinnbarkeit and penetration of spermatozoa into cervical mucus did not differ. In conclusion, the best time for AI is when viscosity is below 20 mm H2O and crystallization is above 3. Viscosity and crystallization could be related to ovulation time, but this needs further investigation. PMID- 10689797 TI - Seroprevalence of Toxoplasma gondii antibodies among domesticated ruminants at AI Qassim Region, Saudi Arabia. PMID- 10689798 TI - The evolutionary genetics of adaptation: a simulation study. AB - It is now clear that the genetic basis of adaptation does not resemble that assumed by the infinitesimal model. Instead, adaptation often involves a modest number of factors of large effect and a greater number of factors of smaller effect. After reviewing relevant experimental studies, I consider recent theoretical attempts to predict the genetic architecture of adaptation from first principles. In particular, I review the history of work on Fisher's geometric model of adaptation, including recent studies which suggest that adaptation should be characterized by exponential distributions of gene effects. I also present the results of new simulation studies that test the robustness of this finding. I explore the effects of changes in the distribution of mutational effects (absolute versus relative) as well as in the nature of the character studied (total phenotypic effect versus single characters). The results show that adaptation towards a fixed optimum is generally characterized by an exponential effects trend. PMID- 10689799 TI - Neutral additive genetic variance in a metapopulation. AB - For neutral, additive quantitative characters, the amount of additive genetic variance within and among populations is predictable from Wright's FST, the effective population size and the mutational variance. The structure of quantitative genetic variance in a subdivided metapopulation can be predicted from results from coalescent theory, thereby allowing single-locus results to predict quantitative genetic processes. The expected total amount of additive genetic variance in a metapopulation of diploid individual is given by 2Ne sigma m2 (1 + FST), where FST is Wright's among-population fixation index, Ne is the eigenvalue effective size of the metapopulation, and sigma m2 is the mutational variance. The expected additive genetic variance within populations is given by 2Ne sigma e2(1-FST), and the variance among demes is given by 4FSTNe sigma m2. These results are general with respect to the types of population structure involved. Furthermore, the dimensionless measure of the quantitative genetic variance among populations, QST, is shown to be generally equal to FST for the neutral additive model. Thus, for all population structures, a value of QST greater than FST for neutral loci is evidence for spatially divergent evolution by natural selection. PMID- 10689800 TI - Clines in polygenic traits. AB - This article outlines theoretical models of clines in additive polygenic traits, which are maintained by stabilizing selection towards a spatially varying optimum. Clines in the trait mean can be accurately predicted, given knowledge of the genetic variance. However, predicting the variance is difficult, because it depends on genetic details. Changes in genetic variance arise from changes in allele frequency, and in linkage disequilibria. Allele frequency changes dominate when selection is weak relative to recombination, and when there are a moderate number of loci. With a continuum of alleles, gene flow inflates the genetic variance in the same way as a source of mutations of small effect. The variance can be approximated by assuming a Gaussian distribution of allelic effects; with a sufficiently steep cline, this is accurate even when mutation and selection alone are better described by the 'House of Cards' approximation. With just two alleles at each locus, the phenotype changes in a similar way: the mean remains close to the optimum, while the variance changes more slowly, and over a wider region. However, there may be substantial cryptic divergence at the underlying loci. With strong selection and many loci, linkage disequilibria are the main cause of changes in genetic variance. Even for strong selection, the infinitesimal model can be closely approximated by assuming a Gaussian distribution of breeding values. Linkage disequilibria can generate a substantial increase in genetic variance, which is concentrated at sharp gradients in trait means. PMID- 10689801 TI - Quantitative genetics in conservation biology. AB - Most of the major genetic concerns in conservation biology, including inbreeding depression, loss of evolutionary potential, genetic adaptation to captivity and outbreeding depression, involve quantitative genetics. Small population size leads to inbreeding and loss of genetic diversity and so increases extinction risk. Captive populations of endangered species are managed to maximize the retention of genetic diversity by minimizing kinship, with subsidiary efforts to minimize inbreeding. There is growing evidence that genetic adaptation to captivity is a major issue in the genetic management of captive populations of endangered species as it reduces reproductive fitness when captive populations are reintroduced into the wild. This problem is not currently addressed, but it can be alleviated by deliberately fragmenting captive populations, with occasional exchange of immigrants to avoid excessive inbreeding. The extent and importance of outbreeding depression is a matter of controversy. Currently, an extremely cautious approach is taken to mixing populations. However, this cannot continue if fragmented populations are to be adequately managed to minimize extinctions. Most genetic management recommendations for endangered species arise directly, or indirectly, from quantitative genetic considerations. PMID- 10689802 TI - Estimating genetic correlations in natural populations. AB - Information on the genetic correlation between traits provides fundamental insight into the constraints on the evolutionary process. Estimates of such correlations are conventionally obtained by raising individuals of known relatedness in artificial environments. However, many species are not readily amenable to controlled breeding programmes, and considerable uncertainty exists over the extent to which estimates derived under benign laboratory conditions reflect the properties of populations in natural settings. Here, non-invasive methods that allow the estimation of genetic correlations from phenotypic measurements derived from individuals of unknown relatedness are introduced. Like the conventional approach, these methods demand large sample sizes in order to yield reasonably precise estimates, and special precautions need to be taken to eliminate bias from shared environmental effects. Provided the sample consists of at least 20% or so relatives, informative estimates of the genetic correlation are obtainable with sample sizes of several hundred individuals, particularly if supplemental information on relatedness is available from polymorphic molecular markers. PMID- 10689803 TI - Artificial selection on phenotypically plastic traits. AB - Many phenotypes respond physiologically or developmentally to continuously distributed environmental variables such as temperature and nutritional quality. Information about phenotypic plasticity can be used to improve the efficiency of artificial selection. Here we show that the quantitative genetic theory for 'infinite-dimensional' traits such as reaction norms provides a natural framework to accomplish this goal. It is expected to improve selection responses by making more efficient use of information about environmental effects than do conventional methods. The approach is illustrated by deriving an index for mass selection of a phenotypically plastic trait. We suggest that the same approach could be extended directly to more general and efficient breeding schemes, such as those based on general best linear unbiased prediction. Methods for estimating genetic covariance functions are reviewed. PMID- 10689804 TI - A classical setting for associations between markers and loci affecting quantitative traits. AB - We examine the relationships between a genetic marker and a locus affecting a quantitative trait by decomposing the genetic effects of the marker locus into additive and dominance effects under a classical genetic model. We discuss the structure of the associations between the marker and the trait locus, paying attention to non-random union of gametes, multiple alleles at the marker and trait loci, and non-additivity of allelic effects at the trait locus. We consider that this greater-than-usual level of generality leads to additional insights, in a way reminiscent of Cockerham's decomposition of genetic variance into five terms: three terms in addition to the usual additive and dominance terms. Using our framework, we examine several common tests of association between a marker and a trait. PMID- 10689805 TI - Estimating the genetic architecture of quantitative traits. AB - Understanding and estimating the structure and parameters associated with the genetic architecture of quantitative traits is a major research focus in quantitative genetics. With the availability of a well-saturated genetic map of molecular markers, it is possible to identify a major part of the structure of the genetic architecture of quantitative traits and to estimate the associated parameters. Multiple interval mapping, which was recently proposed for simultaneously mapping multiple quantitative trait loci (QTL), is well suited to the identification and estimation of the genetic architecture parameters, including the number, genomic positions, effects and interactions of significant QTL and their contribution to the genetic variance. With multiple traits and multiple environments involved in a QTL mapping experiment, pleiotropic effects and QTL by environment interactions can also be estimated. We review the method and discuss issues associated with multiple interval mapping, such as likelihood analysis, model selection, stopping rules and parameter estimation. The potential power and advantages of the method for mapping multiple QTL and estimating the genetic architecture are discussed. We also point out potential problems and difficulties in resolving the details of the genetic architecture as well as other areas that require further investigation. One application of the analysis is to improve genome-wide marker-assisted selection, particularly when the information about epistasis is used for selection with mating. PMID- 10689806 TI - Linkage disequilibrium mapping of molecular polymorphisms at the scabrous locus associated with naturally occurring variation in bristle number in Drosophila melanogaster. AB - We evaluated the hypothesis that the Drosophila melanogaster second chromosome gene scabrous (sca), a candidate sensory bristle number quantitative trait locus (QTL), contributes to naturally occurring variation in bristle number. Variation in abdominal and sternopleural bristle number was quantified for wild-derived sca alleles in seven genetic backgrounds: as homozygous second chromosomes (C2) in an isogenic background, homozygous lines in which approximately 20 cM including the sca locus had been introgressed into the isogenic background (sca BC), as C2 and sca BC heterozygotes and hemizygotes against a P element insertional sca allele and a P-induced sca deficiency in the same isogenic background, and as sca BC heterozygotes against the wild-type sca allele of isogenic strain. Molecular restriction map variation was determined for a 45 kb region including the sca locus, and single-stranded conformational polymorphism (SSCP) was examined for the third intron and parts of the third and fourth exons. Associations between each of the 27 molecular polymorphisms and bristle number were evaluated within each genotype and on the first principal component score determined from all seven genotypes, separately for each sex and bristle trait. Permutation tests were used to assess the empirical significance thresholds, accounting for multiple, correlated tests, and correlated markers. Three sites in regulatory regions were associated with female-specific variation in abdominal bristle number, one of which was an SSCP site in the region of the gene associated with regulation of sca in embryonic abdominal segments. PMID- 10689807 TI - Mapping quantitative trait loci for murine growth: a closer look at genetic architecture. AB - Over 20 years ago, D. S. Falconer and others launched an important avenue of research into the quantitative of body size growth in mice. This study continues in that tradition by locating quantitative trait loci (QTLs) responsible for murine growth, such as age-specific weights and growth periods, and examining the genetic architecture for body weight. We identified a large number of potential QTLs in an earlier F2 intercross (Intercross I) of the SM/J and LG/J inbred mouse strains. Many of these QTLs are replicated in a second F2 intercross (Intercross II) between the same two strains. These replicated regions provide candidate regions for future fine-mapping studies. We also examined body size and growth QTLs using the combined data set from these two intercrosses, resulting in 96 microsatellite markers being scored for 1045 individuals. An examination of the genetic architecture for age-specific weight and growth periods resulted in locating 20 separate QTLs, which were mainly additive in nature, although dominance was found to affect early growth and body size. QTLs affecting early and late growth were generally distinct, mapping to separate chromosome locations. This QTL pattern indicates largely separate genetic and physiological systems for early and later murine growth, as Falconer suggested. We also found sex-specific QTLs for body size with implications for the evolution of sexual dimorphism. PMID- 10689808 TI - Testing the correspondence between map positions of quantitative trait loci. AB - There are several instances in which quantitative trait locus (QTL) mapping experiments have been independently carried out for similar traits in different laboratories. We develop a permutation test of the correspondence between the test statistics obtained from genome-wide QTL scans in two such experiments to test whether the same QTLs are segregating in the experimental pair. In simulations, we show that the permutation test has the desired properties if chromosomes are of equal length, but bias can occur if chromosomes are of unequal length, a problem connected with autocorrelation of test statistic values. We apply the test to data from three recent mouse body weight QTL mapping experiments. The results from the test are non-significant, and imply a lack of overall concordance between the QTLs that were segregating in these experiments. PMID- 10689809 TI - The genetic basis of inbreeding depression. AB - Data on the effects of inbreeding on fitness components are reviewed in the light of population genetic models of the possible genetic causes of inbreeding depression. Deleterious mutations probably play a major role in causing inbreeding depression. Putting together the different kinds of quantitative genetic data, it is difficult to account for the very large effects of inbreeding on fitness in Drosophila and outcrossing plants without a significant contribution from variability maintained by selection. Overdominant effects of alleles on fitness components seem not to be important in most cases. Recessive or partially recessive deleterious effects of alleles, some maintained by mutation pressure and some by balancing selection, thus seem to be the most important source of inbreeding depression. Possible experimental approaches to resolving outstanding questions are discussed. PMID- 10689810 TI - Properties of spontaneous mutations affecting quantitative traits. AB - Recent mutation accumulation results from invertebrate species suggest that mild deleterious mutation is far less frequent than previously thought, implying smaller expressed mutational loads. Although the rate (lambda) and effect (s) of very slight deleterious mutation remain unknown, most mutational fitness decline would come from moderately deleterious mutation (s approximately 0.2, lambda approximately 0.03), and this situation would not qualitatively change in harsh environments. Estimates of the average coefficient of dominance (h) of non-severe deleterious mutations are controversial. The typical value of h = 0.4 can be questioned, and a lower estimate (about 0.1) is suggested. Estimated mutational parameters are remarkably alike for morphological and fitness component traits (excluding lethals), indicating low mutation rates and moderate mutational effects, with a distribution generally showing strong negative asymmetry and little leptokurtosis. New mutations showed considerable genotype-environment interaction. However, the mutational variance of fitness-component traits due to non-severe detrimental mutations did not increase with environmental harshness. For morphological traits, a class of predominantly additive mutations with no detectable effect on fitness and relatively small effect on the trait was identified. This should be close to that responsible for standing variation in natural populations. PMID- 10689811 TI - Role of growth hormone in the genetic change of mice divergently selected for body weight and fatness. AB - To elucidate the involvement of growth hormone (GH) in the genetic change produced by long-term selection in growth and fatness, a 'GH knock-out study' on over 900 mice was undertaken. Lines used had been selected for more than 50 generations for high (PH) and low (PL) body weight (initially protein mass) at 70 d(ays) and for high (F) and low fat content (L) at 98 d, producing a 3-fold difference in body weight and a 5-fold difference in fat content. GH deficiency was achieved by repeated backcrossing into each line a recessive mutant gene (lit) which has a defective GH releasing factor receptor. In the absence of GH, the P lines still differ in body weight (21 d to 98 d): e.g. at 98 d homozygous lit/lit: PH = 24.2 g, PL = 10.0 g; wild-type (wt): PH = 57.4 g, PL = 18.7 g. The effect of the GH deficiency on body weight (untransformed) was very much larger in the PH than in the PL line, but the interaction was much smaller, although still significant, on the log scale. This indicates that changes in the GH system contribute only a small part of the selection response in growth. GH deficiency increased fat percentage in all lines (including P), especially in males (99 d, males lit/lit: F = 26.4%, L = 6.9%; wt: F = 22.0%, L = 4.8%; females: 20.2%, 5.2%, 20.7%, 3.0%) with significant genotype x line and genotype x sex interactions. The interactions between the effects of the lit gene and the genetic background were, however, relatively small compared with these main effects and again indicate that other systems contributed most of the selection response. PMID- 10689812 TI - [BCG vaccination in the world]. AB - BCG vaccination programme and BCG vaccination coverage in the world were summarized mainly based on the published informations from official organizations, such as World Health Organization (WHO), International Union Against Tuberculosis and Lung Disease (IUATLD) and Centers for Disease Control and Prevention (CDC). From this review, we can see how widely BCG has been used for the prevention of tuberculosis in the world. In most of the developing countries, especially in Africa, the Americas, and Pacific Region, BCG vaccination is carried out to newborn babies soon after birth by intradermal injection according to the recommendations from WHO, but some of the developing countries in Asia and Europe have their own modified BCG vaccination programmes. In economically developed countries, BCG vaccination programme has been established according to the tuberculosis status of each countries. Some countries have general vaccination policy, and other countries have selected vaccination policy, but there is no country where BCG vaccination is not carried out at all. Among G8 contries, as representatives of the economically developed countries, Japan, United Kingdom, France and Russian Federation have BCG general vaccination policy for the specified age group. In these 4 countries revaccination (s) of BCG are still carried out. In Germany, some provinces have general vaccination policy and some others have selected vaccination policy. In the United States of America, BCG vaccination is recommended to selected high risk infants and health care workers by CDC. There are many debates as for the efficacy and safety of BCG vaccination, and the development of new vaccine better than BCG has been actively discussed and some encouraging results in animal models have been reported from several laboratories. But, there is almost no possibility to be able to use a new vaccine in the routine practice within a couple of years. From the practical point of view, therefore, the operational researches for the better and more appropriate usage of BCG are equally important and more practical than the researches for the development of new vaccines. PMID- 10689813 TI - [Effect of serotype specific glycopeptidolipid (GPL) isolated from Mycobacterium avium complex (MAC) on phagocytosis and phagosome-lysosome fusion of human peripheral blood monocytes]. AB - Mycobacterium avium complex (MAC) is a typical intracellular parasite similar to M. tuberculosis and is one of the most important pathogens that coinfects AIDS patients. Attention has been focused on M. avium infection causing immunosuppression of hosts. Specific serotype-subspecies such as 1, -4 or -8 serotypes can be isolated frequently in humans infected with HIV. Furthermore, the prognosis after infection differs depending on the serotype. Serotype-4 in general shows unfavourable prognosis, while serotype-16 yields rapid recovery. Therefore, we have been interested in the immunomodifying activity of the surface glycopeptidolipid (GPL) antigen. However, no information has been available to date dealing on the virulent factor of MAC that is directly related with intracellular bactericidal activity. Recently, we have tried to test the effect of various GPLs purified from MAC on phagocytic processes of human peripheral blood monocytes (PBMC). We have used GPL-coated heat-killed staphylococcal cells to be phagocytosed by PBMC, and phagosome-lysosome fusion (P-L fusion) was estimated by the acridine orange staining of fused vesicles and bacteria. Results showed strong promotion of phagocytosis and marked inhibition of P-L fusion by serotype-4 GPL, while neither promotion of phagocytosis nor inhibition of P-L fusion in phagocytic cells were shown by serotype-16 GPL. Serotype-8 GPL showed concomitant stimulation of both phagocytosis and P-L fusion. These effects may be due to some unknown interaction between specific carbohydrate chain and organella membranes and serotype-4 GPL may be one of the possible virulent factors in MAC. Comparison with known possible virulent factors such as trehalose 6,6'-dimycolate (TDM), trehalose 6-monomycolate (TMM), glucose 6-monomycolate (GM) or sulfatide was also reported. PMID- 10689814 TI - [Isolation of Mycobacterium avium complex from the "24-hour bath"]. AB - The "24-HOUR BATH" is an apparatus which circulates the bath water, keeps it clean and warm, and makes it possible to take a bath at any time during the day or night. It consists of apparatus for cleaning (sponge or mesh filter and filter material), heating (ceramic heater), and sterilizing (UV lamp). Recently, three cases of skin disease due to M. avium infection in private homes, in which "24 HOUR BATH" water was suspected to be the source of infection, have been reported. We attempted to isolate M. avium complex from the water (32 specimens), sponge filter (29 specimens), and filter material (32 specimens) of the "24-HOUR BATH". One hundred-ml samples of bath water, and 50-ml samples of rinse from a sponge filter or filter material were centrifuged at 3000 rpm for 20 min. Sediment was suspended in distilled water and a smear was prepared, and then digested and decontaminated with 2% sodium hydroxide. The processed specimens were cultured on 2% Ogawa medium containing ofloxacin (1 microgram/ml) and ethambutol (2.5 micrograms/ml) for 8 weeks at 37 degrees C. Positive smears were 3 (9.4%), 25 (86.2%) and 25 (78.1%) specimens from the water, sponge and filter material, respectively. A few bacterial clumps were observed, especially in the sponge specimens. The number of positive culture was 5 (15.6%), 24 (82.8%) and 25 (78.1%) from the water, sponge and filter material, respectively. Among them the number of Runyon's Group III-positive cultures was 5 (100%), 22 (91.7%) and 20 (80%) in the water, sponge, and filter material specimens, respectively. In most cases, cultures were positive for both the sponge and filter material specimens. All of the Group III mycobacteria were smooth, grew at 28, 37, 42, and 45 degrees C, negative for niacin, nitrate reductase, semiquantitative catalase, urease and Tween80 hydrolysis, and positive for 68 degrees C catalase. All of the strains reacted with M. avium complex AccuProbe and M. avium AccuProbe, but none of the strains reacted with M. intracellulare AccuProbe. Therefore, all the Group III isolates were identified as M. avium by the culture, biochemical and genetical characteristics. PMID- 10689815 TI - [Two-step tuberculin skin test in nurse students and hospital employees]. AB - Booster phenomenon (recall effect) of tuberculin skin test, which disturbs diagnosis of tuberculous infection, is prevalent among BCG vaccinated population. We retested 34 nurse students whose initial tuberculin reaction was smaller than 30 mm by erythema (Group A) and 53 hospital employees whose initial reaction was smaller than 20 mm by erythema (Group B). Among the people whose diameter of erythema was less than 10 mm by the first test, 88 percent (8/9) of group A and 43% (6/14) of group B showed reaction 10 mm < or = by erythema and among those whose induration was < 10 mm, 54% (6/11) of group A and 48% (12/25) of group B showed reaction 10 mm < or = in the second testing. Mean and standard deviation of [the difference between the diameter of the 2nd and the 1st testing] was +7.3 +/- 11.8 mm in group A, +9.8 +/- 11.1 mm in group B by erythema and +2.6 +/- 5.9 mm in group A, +2.9 +/- 5.1 mm in group B by induration. These results indicate that booster phenomenon is highly prevalent among the tested group and there can be no appropriate criteria to distinguish new infection and booster phenomenon. Though two-step tuberculin skin test is recommended to get rid of booster phenomenon. Only a little is known about the value of this test to diagnose new infection in Japanese population, majority of whom being BCG vaccinated. Further investigations are required to apply two-step tuberculin skin test for diagnosis of new infection among hospital employees and health care workers in Japan. PMID- 10689816 TI - [A case of pulmonary tuberculosis complicated with subcutaneous phaeohyphomycosis]. AB - A 78-year-old male was admitted to our hospital because of fever, sputum and cough. Chest X-ray showed infiltrative shadows in the right lung field. Smears of his sputum were positive for acid-fast bacilli. We found multiple subcutaneous abscesses on the right distal forearm. Microscopic examination of skin biopsy specimens revealed granulation tissues with the proliferation of epitheloid cells with the scattering infiltrations of neurophils, giant cells and histiocytic cells. The examination of the PAS stained specimen revealed fungal elements and a black fungus, Exophiala jeanselmei, was isolated by the cultures of pus from the abscess. He was diagnosed as pulmonary tuberculosis complicated with subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei and was successfully treated with anti-tuberculosis drugs and anti-fungal agent, 5-fluorocytosine. PMID- 10689817 TI - [Endoscopic approach to pulmonary diseases: Transbronchial needle aspiration]. AB - Transbronchial needle aspiration (TBNA) is a bronchoscopic technique to obtain cytologic and histologic specimen from deep site of bronchial wall. We investigated the utility and safety of TBNA in 200 patients who had mass lesions in the lung or in the mediastinum. 101 patients had submucosal or peribronchial malignant lesions (central malignancy) and TBNA was the only diagnostic method in 28 patients (28%). 39 patients had peripheral malignant lesions (peripheral malignancy) and TBNA was the only diagnostic method in 12 patients (31%). The other 60 patients had benign lesions and TBNA was diagnostic in only 5 patients (8%); bronchogenic cyst in 2, tuberculous lymph adenitis in 1, parathyroid adenoma in 1 and lung abscess in 1. In central malignancy, the yield of TBNA as exclusive means of diagnosis was higher for mediastinal tumor than for lung cancer. In peripheral malignancy it was higher for metastatic lung tumor than for primary lung tumor. In order to stage patients of lung cancer, we sampled 39 lymph nodes and 21 of them were proved to be positive. TBNA was thought to be of greatest value in the diagnosis of peritracheal mediastinal tumor and peribronchial metastatic lung tumor and in the staging of lung cancer. We used 19 gauge transbronchial histology needle in 8 patients and 2 cases were diagnostic. Low diagnostic yields were probably due to the lack of our experience and it was expected that training on this technique would increase diagnostic utility of the histology needle. No significant complications occurred and all patients tolerated TBNA well. PMID- 10689818 TI - [Endoscopic approach to pulmonary diseases: Bronchoscopy for critical respiratory care in neonates, infants, and children]. AB - Of the 605 pediatric patients admitted to our intensive care unit during the past 6 years, 90 underwent 380 bronchoscopies for diagnostic and therapeutic purposes. The indications for bronchoscopy were atelectasis/retention of airway secretion (n = 52), pneumonia (n = 31), airway bleeding (n = 14), pulmonary edema (n = 11), tracheobronchomalacia/airway stenosis (n = 11), and airway foreign body/aspiration (n = 7). Visualization of the airway was helpful for the diagnosis of respiratory problems; in 9 infants, bronchoscopy revealed tracheobronchial stenosis which other diagnostic modalities failed to detect. In 25 of the 31 patients with pneumonia, specimens taken by bronchoscopy were positive for specific pathogens. Bronchoscopy also proved to have significant therapeutic value, especially for airway cleaning; bronchial suctioning resulted in immediate reexpansion of the collapsed lung in 25 of 34 cases of physiotherapy resistant atelectasis. A rigid bronchoscope was used to remove airway foreign body in 2 cases. Prototype channeled-ultrathin fiberscopes were utilized in 99 of 380 bronchoscopies. No complications were noted except for severe hypoxia and bradycardia in one infant. We conclude that bronchoscopy is a safe and useful modality for the critical respiratory care of infants and children. PMID- 10689819 TI - [Endoscopic approach to pulmonary diseases: Clinical utility of medical thoracoscopy in diagnosis of pleural diseases]. AB - Thoracoscopy is useful for diagnosis of a number of lung diseases. We report our recent experience of medical thoracoscopy performed under local anesthesia in 142 cases. Of 124 patients with pleural effusion, 46 had pleuritis carcinomatosa, 11 had pleuritis tuberculosa, and 10 had malignant mesothelioma. We evaluated the utility of thoracoscopic observation and pleural biopsy in these three diseases. Almost of patients with malignant pleural effusion initially undiagnosed by the cytology of pleural effusion were diagnosed by thoracoscopy. Especially in malignant mesothelioma, thoracoscopy allowed accurate diagnosis. No serious complication was observed. Since medical thoracoscopy under local anesthesia is a rapid, easy, safe, and well-tolerated procedure with an excellent diagnostic yield, it is recommended as a diagnostic procedure for cases with pleural effusion. PMID- 10689820 TI - [Endoscopic approach to pulmonary diseases: Usefulness of the mediastinoscopy]. AB - The purposes of this study are to show the diagnostic values and the role of the mediastinoscopy for the respiratory diseases. From 1971 to 1998, mediastinoscopy were performed on 1664 patients admitted to our hospital with respiratory diseases. For the superior mediastinal diseases, mediastinal tumor and lymphadenopathies without cancer, two or three samples were obtained by mediastinoscopy. For lung cancer, biopsy was routinely performed at the 6 nodal stations, right and left paratracheal (#2), right and left tracheobronchial (#4), pretracheal (#3), and subcarinal (#7) lymphnodes. From 1994, we have used video mediastinoscopy, which was combined with scope and TV-camera. Using video mediastinoscopy, many staffs could observe the mediastinal findings on TV-monitor during mediastinal manipulation. The positive findings were observed in 17% (221/1299) for lung cancer, 100% (32/32) for sarcoidosis, 100% (2/2) for malignant lymphoma, 65% (11/17) for mediastinal tumor, 9.8% (13/132) for pulmonary tuberculosis. The positive rate according to the histological types of lung cancer were 20.5% (148/721) for adenocarcinoma, 9.4% (39/415) for squamous cell carcinoma, 31.6% (24/76) for small cell carcinoma, 21.3% (10/47) for large cell carcinoma. Complications developed in a total of 3.6%, and these were bronchial arterial damage(1.8%), recurrent nerve paralysis(0.7%), azygos vein damage (0.4%), pleural rupture(0.4%), superior vena cava damage(0.2%) and tracheal laceration(0.1%). However, there were no severe complications and operative deaths in this series. Mediastinoscopy is a minimal invasive and safety surgical procedure that is widely used as a diagnostic method for investigating the superior mediastinum, mediastinal tumor and lymphadenopathies. It is useful for obtaining histological diagnosis, as well as for staging lung cancer. Video mediastinoscopy is more safety and educational, because many staffs could observe the findings. PMID- 10689821 TI - Leprosy elimination campaigns (LEC). PMID- 10689822 TI - Guidelines for carrying out leprosy elimination campaigns 1996. WHO, Action Programme for the Elimination of Leprosy. AB - A Leprosy Elimination Campaign (LEC) is an initiative which aims to detect leprosy cases, particularly the more serious ones referred to as 'cases of consequence', that remain undetected in the community, and to treat them with MDT. This will subsequently reduce the delay in managing such cases in the community and ensure that the existing health services are able to treat them. It is a time-bound, one-time activity. Leprosy of consequence is defined as cases with more than five skin lesions and skin smear positive cases. Such cases are regarded as being of consequence because they act as a major source of infection in the community and are either already disabled or at high risk of becoming disabled. LEC is a focused combination of activities which includes: orientation courses for local health workers and volunteers; community awareness creation activities; case-finding and treating every detected case immediately with MDT. These campaigns are to be carried out primarily in endemic regions where the prevalence and new case detections are high and where the gap between estimated and registered cases is large. It is a national activity, implemented by the national staff with technical cooperation from WHO and other agencies. PMID- 10689823 TI - Leprosy elimination--sprint or marathon? PMID- 10689824 TI - Progress towards elimination of leprosy as a public health problem in India and role of modified leprosy elimination campaign. AB - India (population 943 million) has seen a highly significant decrease in the prevalence of leprosy since the introduction of multi-drug therapy (MDT) in 1981. From a prevalence rate of 57/10,000 of the population in March 1981, the figure has declined to 5.2/10,000 in March 1999. This was possible due to the creation of a completely vertical (specialized) infrastructure for leprosy control in the 218 endemic districts of the country and skeleton vertical staff in the remaining districts, coupled with the recruitment of additional staff on contract basis to provide MDT through vertical staff in endemic districts and mobile treatment units in the moderate and low endemic districts. Despite all efforts, however, new case detection has not shown a decline over the last 14 years due to the presence of hidden (and undiagnosed) cases. Therefore, in order to intensify and hasten progress towards elimination (less than 1 case per 10,000 of the population) in the whole country, it was decided to implement a massive leprosy elimination campaign (LEC) in all the States/Union Territories (UTs). The reports of 22 States/UTs indicate that 415 out of the total of 490 districts in the country were covered by modified LEC (MLEC), with 85% coverage of the population. The campaign used in India was modified from the pattern previously described by the World Health Organization. The detection of hidden or suspected cases took place within a short, intensive period of 6-7 days and relied heavily on house-to house searches by General Health Care staff trained in leprosy detection and confirmation was made by appropriately trained staff. This MLEC received widespread Government and public support, resulting in the detection of 454,290 hidden cases of leprosy, whilst providing training to a large number of General Health Care staff and volunteers and creating widespread awareness about leprosy and the availability of treatment free of charge for all cases. This programme proved to be one of the most successful health care interventions undertaken in India in recent years, particularly in the states of Bihar and Orissa. Although a few states in India are unlikely to reach the current WHO goal of elimination before end of the year 2000, the results of the MLEC strongly support the possibility that elimination levels will be achieved in the majority of states by the end of the year 2000 and at national level by the end of the year 2002. PMID- 10689825 TI - Modified leprosy elimination campaign (MLEC) in the State of Orissa, India. AB - As part of a country-wide modified leprosy elimination campaign (MLEC) carried out in 21 selected States in India in 1998, the State of Orissa launched activities in early January of that year, during which 28.9 million people were examined, giving 85% coverage of the enumerated population. Using general health care staff and volunteers, 416,604 suspect cases were identified and 62,804 of these were confirmed as leprosy by experience observers. The period of intensive search activity lasted 1 week only, but this was preceded by several months of community mobilization and involvement, health education, training of government and voluntary staff, media messages and the involvement of all relevant health departments, officials and politicians. Both this and the intensive search period were characterized by a high level of interest and cooperation by all concerned. The total of new cases detected and put on treatment (multi-drug therapy; MDT) during the period of only 7 days was approximately equal to that which, on routine population survey by the leprosy services, would be recorded over a period of 2 years. The MLEC in Orissa is judged to have been not only an historic step forward in the control of leprosy in a State previously classified as highly endemic for leprosy, but also one of the most successful State health interventions ever mounted. In the 5 months after completion of the campaign, the voluntary reporting rate increased from 50 to 90%. As a direct result of the campaign, facilities for the diagnosis and treatment of leprosy are now available daily in an additional 1639 institutions, over and above those in existence before the campaign was launched. The achievements in terms of detecting hidden (and thus undiagnosed and untreated) cases exceeded the outset predictions, underlining the importance of continued vigilance and the need to maintain involvement of general health care staff. It is anticipated that the rise in prevalence due to the addition of 62,884 cases will be reduced by the implementation of MDT by 80% by about March 1999. Overall the results of the MLEC in Orissa strongly support the likelihood that an elimination level of less than 1 case per 10,000 of the population will be reached in this State by the year 2000. PMID- 10689826 TI - Leprosy elimination campaign in a metropolitan leprosy project, Bombay, India. PMID- 10689827 TI - Pace of leprosy elimination and support teams in Bihar state, India. AB - Despite the extensive implementation of multiple drug therapy (MDT) in most leprosy-endemic countries world-wide since 1982, bringing about a remarkable reduction in prevalence, there are still regions at the sub-national level where the implementation of MDT remains difficult. The state of Bihar (population 86.3 million) in India is a good example of such a region. Previously rated as one of the most highly endemic states, it still contributes about 21% of the total caseload in India and about 12% of the global caseload. For various reasons, case finding and drug treatment have lagged behind the progress made in most other states in the country and in 1996, the Damien Foundation India Trust (DFIT) volunteered technical support to increase the pace of elimination. Sixteen out of the 39 districts in the state were allocated, with a population of 41.8 million. Support teams, including a Medical Advisor and a Non-Medical Supervisor, both with over 10 years experience of leprosy work and control programmes, were provided to assist and work alongside government staff in case detection, treatment delivery, case-holding and discharge in their respective areas of operation. New case detection by intensive survey increased by 394% and total new case detection by 226% during the year 1996-1997, with similar trends in the following year. Striking improvements were also observed in MDT coverage, treatment regularity, monitoring and discharge of patients and in the training of local staff. This collaboration between a non-government agency (DFIT) and the staff of the National Leprosy Eradication Programme in 16 out of 39 districts in the State of Bihar has clearly been extremely successful. Similar approaches in the remaining districts of Bihar, and in other parts of India, where the infrastructure is available but inadequate, may contribute significantly to achieving the elimination goal at national and sub-national levels. PMID- 10689828 TI - Modified leprosy elimination campaign in Mumbai (Bombay), India--a report. AB - With appropriate planning and preparation, a modified leprosy elimination campaign (MLEC) was undertaken in Brihan Mumbai (Bombay), which has a population of around 11 million. For the campaign, 4879 non-leprosy paramedical and non medical personnel were trained and utilized as searchers. The MLEC revealed 1410 new leprosy cases, with a new case detection rate of 1.83/10,000. Over 80% of all cases detected were either single-lesion or paucibacillary (PB), and thus of limited significance with regard to transmission. Further efforts are required to detect and treat cases of consequence (those with more than five lesions and those with positive skin smears) and to identify reservoirs of infection. PMID- 10689829 TI - Leprosy elimination campaigns: the Nigerian experience. PMID- 10689830 TI - Report of the national leprosy elimination campaign (NLEC) of Bangladesh, 1999. AB - A national leprosy elimination campaign (NLEC) was implemented country-wide in all the 64 districts of Bangladesh for 6 days from 7 to 12 February 1999. The campaign was jointly funded by the Government of Bangladesh (GOB)/World Bank (US$250,000) and the remaining US$381,000 was provided by other international non governmental organizations (NGOs). A total of 44,400 health workers and community volunteers were directly involved in the campaign. In all, 60,878 suspected leprosy cases were identified during the campaign, of whom 31,433 were examined and 2435 were confirmed as leprosy cases. The remaining suspects are expected to be examined within the next 2 months. Details of the new cases detected are given in Table 8. The impact of NLEC has been significant, the number of cases detected during NLEC being 20% of the annual case detection in 1998. About 52% of the total population were directly contacted through a rapid house-to-house survey and over 90% of the population was targeted through extensive use of electronic/print media and various information, education and communication (IEC) activities. PMID- 10689831 TI - Leprosy elimination campaign (LEC) in Myanmar, 1997 to May 1999. PMID- 10689832 TI - Leprosy elimination campaign (LEC) in the Philippines. PMID- 10689833 TI - The Dharavi story--saga of LECs over 2 decades. PMID- 10689834 TI - Future scope and expectations: why, when, and how LECs should continue. AB - There is a strong case to continue to use LEC approaches, as they are a comprehensive and cost effective means of delivering the key elements of leprosy control. LECs should be conducted when there is evidence of large numbers of hidden cases. Probably a minimum of two LECs is required but where large number of new cases continue to be detected they could be run on an annual basis. The methodology of LECs needs to be improved through experience, evaluation and from LECs conducted elsewhere; feedback from the community is also important. There is room to improve all aspects of LECs: planning, training, education, diagnosis and treatment completion. PMID- 10689835 TI - [Propofol potentiates the neuromuscular blocking effects of vecuronium in man]. AB - The possible interaction between vecuronium and propofol has been investigated in 40 healthy (ASA I-II) patients. They were randomly allocated to two groups according to the method of anesthesia; continuous propofol infusion group (propofol) and droperidol and fentanyl group (control). The electromyographic response of abductor digiti minimi was monitored at 20-s interval after train-of four stimulations of the ulnar nerve. The ED50 and ED95 (dose required to produce a 50% and 95% depression of twitch tension, respectively,) of vecuronium in the propofol group (n = 20) were 29.4 +/- 0.5 and 56.6 +/- 2.1 micrograms.kg-1 (mean +/- SEM), and in the control group (n = 20), 36.7 +/- 1.8 and 73.6 +/- 5.2 micrograms.kg-1, respectively. Under propofol anesthesia, the cumulative dose response curves of vecuronium were shifted to the left when compared with control ED50 and the slope showed that propofol had potentiated the action of vecuronium. PMID- 10689836 TI - [Lack of communication between anesthesiologists and surgeons: comparison of questionnaire survey among anesthesiologists with that among surgeons concerning pre-anesthetic evaluation of surgical patients]. AB - We conducted a survey to examine surgeons' opinions and criticisms of patient evaluations done by anesthesiologists prior to surgery. We sent questionnaires to 117 departments of general surgery in Japanese university hospitals. We received answers from 77% of the departments. We analyzed their answers, and compared the answers with those from a similar survey done in 1995 by Japan Society of Anesthesiology, one in which anesthesiologists were asked to evaluate themselves. Our most significant findings were as follows. (1) Although most of surgeons were satisfied with their own preanesthetic evaluation of their patients, 30% of the departments reported postponement of surgery due to the need of further examination during recent 2 months and the occurrence of peri-operative myocardial infarction during recent 2 years, (2) The 1995 survey indicated that 46 percent of anesthesiology departments had explained the major perioperative risk, but a half of the 46% had done so without discussing the risk with responsible surgeons. Furthermore, the present survey showed that only 17% of the surgery departments had been aware of such explanation done by anesthesiologists. (3) One-third of the anesthesiology departments did not document the text of the preanesthetic explanation to patients. In our opinion, the final responsibility for the patient's peri-operative care is primarily the surgeon's at present in Japan, although each specialist including the anesthesiologist and the cardiologist should share the responsibility. If the anesthesiologist explain the major peri-operative risk to the patient without first obtaining the surgeon's permission to do so, the patient may become confused about who is responsible for his or her care. It should be made clear to the patient what responsibility each doctor has. It is also important that all explanations given to a patient and the consent to anesthesia given by a patient be properly documented. Japanese anesthesiologists and surgeons need to work far more closely together with regard to pre-anesthetic evaluation and explaining patients about their peri-operative risk. PMID- 10689837 TI - [Factors influencing the level of spinal anesthesia: (I). Characteristics of anesthetic solutions]. AB - The practitioner of spinal anesthesia needs to know the minimum block he or she can expect in order to guarantee the adequate anesthesia to perform a given operation. At the same time, the anesthesiologist needs to know the maximum extent of block, for which he or she must be prepared to avoid being caught unaware. The height of sensory block is determined by the cephalad distribution of the local anesthetic in the cerebrospinal fluid and uptake by neuronal tissue in sufficient amounts to produce the block. Out of many factors that have been considered to affect the distribution, this paper discusses characteristics of anesthetic solutions. It has been suggested that solutions that are marginally hyperbaric can safely produce relatively consistent blocks with an extent that is suitable for many operations performed under spinal anesthesia. PMID- 10689838 TI - [A case of water intoxication during transcervical resection of the uterine myoma]. AB - A 28-year-old woman underwent trans-cervical resection (TCR) of the uterine myoma. She had no history of complications except for anemia. Anesthesia was maintained with inhalation anesthesia. After 120 min from introducing the resectoscope, her serum sodium dropped to 86.1 mEq.l-1. But her heart rate and the systolic arterial pressure were stable. She was treated with steroid, mannitol, isotonic saline, and 7% NaHCO3. Soon after the end of the operation, she recovered consciousness. At two hours postoperatively, her serum sodium was 119.6 mEq.l-1. On the 1st postoperative day, her serum sodium returned to the normal range (137 mEq.l-1). We should be aware of asymptomatic water intoxication during TCR. PMID- 10689839 TI - [Anesthetic management of a patient with hemophilia A for left modified Blalock Taussig shunt]. AB - We gave anesthesia to a patient with hemophilia A for left modified Blalock Taussig shunt. The patient was a twenty five-day-old boy with pulmonary atresia. We performed the bolus injection test of factor VIII concentrate in preoperative period. His factor VIII activity increased from 9.3 to 113.3% after a bolus injection of 165 units. To keep his factor VIII activity above 80% in perioperative period, a bolus of 125 units of recombinant factor VIII concentrate was injected at anesthesia induction, 125 units 2 hours after the start of the operation, and 125 units 6 hours after the end of the operation. Factor VIII activity 2 hours after anesthesia induction increased only 37.8%, and we had to infuse recombinant factor VIII concentrate additionally. We measured factor VIII activity during operation, and he finally received total of 415 units of factor VIII concentrate. Hydroxyethyl starch infusion, blood transfusion and bleeding in the perioperative period might have caused the factor VIII activity to decrease beyond our expectation. We should infuse factor VIII concentrate properly measuring the factor VIII activity during this operation. PMID- 10689840 TI - [The anesthetic management for elective or emergent cesarean section in patients with intracranial arteriovenous malformation]. AB - Subarachnoid hemorrhage secondary to ruptured intracranial arteriovenous malformation (AVM) during pregnancy, although rare, is a grave complication. We experienced 3 patients with AVM for cesarean section. Case 1: A 24-year-old woman suffered sudden vomiting and headache during the 22nd week of her first pregnancy. She was diagnosed as having the intracranial hemorrhage due to AVM. Because the patient was bleeding again at 29th week of pregnancy, emergency operation was performed. Her neurological symptom improved. Cesarean section was performed under general anesthesia at 34th week of pregnancy. Case 2: A 42-year old woman of her first pregnancy had past history of subarachnoid hemorrhage due to AVM at the ages of 23, 28, 29 and 36. The malformation was not corrected surgically. Her neurological status was normal. Cesarean section was performed under spinal anesthesia. Case 3: A 29-year-old woman suffered sudden hemiplegia, vomiting and headache during the 40th week of her first pregnancy. She was diagnosed as having intracranial hemorrhage. Cesarean section immediately followed by the removal of an intra cranial hematoma under general anesthesia. Better perinatal outcome is expected when AVM rerupture is prevented by first performing cesarean section. PMID- 10689841 TI - [Iatrogenic extrapleural hematoma]. AB - We encountered a rare case of complications at the time of central venous catheterization due to extrapleural hematoma. A 71-year-old woman was scheduled to undergo subtotal gastrectomy. After introduction of general anesthesia, a CVP catheter was inserted from the right jugular vein, but it was removed intraoperatively, because of poor dropping of the infusion fluid. A few minutes later, the blood pressure started to decrease. We considered that this symptom was derived from the surgical procedure, and rapid blood transfusion associated with administration of a vasopressor was performed. Postoperative chest X ray revealed poorly delineated right lung field, and hemothorax was suspected. However thoracic drainage resulted in an extremely small amount of blood-like fluid. The abnormal defect in the right pulmonary field was found to be an extrapleural hematoma by thoracic CT on the first postoperative day. The hematoma was reduced by subsequent management in 7 days, and the patient was discharged from the ICU without any further complications. PMID- 10689842 TI - [Malfunctioning of cerebral function monitors in three cases of carotid endarterectomy]. AB - In three patients, EEG, jugular venous oxygen saturation (Sjvo2) and near infrared spectroscopy (NIRS) were monitored to detect cerebral ischemia during carotid endarterectomy. In all cases, no changes in Sjvo2 and NIRS were observed during carotid artery occlusion, but in two patients EEG showed changes when carotid artery was clamped. It is important to know the precise mechanism of cerebral monitors to assess the cerebral ischemia in patients with preexisting neurological disorder during carotid endarterectomy. PMID- 10689843 TI - [Anesthetic management in a patient with severe acute pancreatitis during pregnancy]. AB - Continuous epidural anesthesia was used in a 34 year-old pregnant woman with acute pancreatitis related to hypertriglyceridemia. She underwent an emergency cesarean section due to severe pancreatitis under spinal anesthesia. After delivery, extended incision was made to examine the pancreas and to perform drainage. Epidural infusion using 1% mepivacaine and buprenorphine was started to reduce pain and improve microcirculation. After starting epidural infusion with other therapies, clinical feature and data improved. This case suggests that reduction of severe pain and improvement of microcirculation are important in therapies of severe pancreatitis. PMID- 10689844 TI - [Heart rate reduction using edrophonium during coronary artery bypass grafting without cardiopulmonary bypass]. AB - Coronary artery bypass grafting without the use of cardiopulmonary bypass (CPB) is performed with increasing frequency. Performing revascularization on a beating heart is technically more demanding than performing revascularization on the arrested heart, especially in high-risk patients. beta-Blockers and calcium channel antagonists have been used for the reduction of heart rate (HR) for the local immobilization of the anastomotic site. However, their negative inotropic actions often lead to serious hypotension. Therefore, we investigated the effect of edrophonium on HR reduction in high-risk patients undergoing CABG without CPB. Ten high-risk patients undergoing CABG without CPB were selected. To reduce HR during anastomosis, edrophonium was administered during the procedure. Systemic blood pressure (sBP), HR, and cardiac index (CI) were measured from the induction of anesthesia to the end of surgery. All surgeries were successfully performed without serious complications. To keep the rate under 60 bpm, edrophonium was administered at the time of anastomosis and this decreased the cardiac index from 2.19 to 1.95, while the sPB was maintained easily over 90 mmHg with the infusion of methoxamine. Edrophonium may be useful for the reduction of HR during coronary anastomosis in high-risk patients undergoing CABG without CPB. PMID- 10689845 TI - [The effect of prophylactic nicorandil infusion to reduce intraoperative myocardial ischemia during abdominal surgery in patients with risk factors of ischemic heart disease]. AB - Three hundred patients who had undergone abdominal surgery with risk factors of ischemic heart disease (IHD), such as hypertension, diabetes mellitus, hyperlipidemia, smoking, advanced age, obesity, familial history, electrocardiographic abnormality, other perivascular disease, and male, were included in this study. Patients older than forty years were included in the study. They were divided into two groups for a randomized double blind trial: nicorandil was administered in one group (group N) and placebo in another (group C), and we investigated the effect of prophylactic nicorandil infusion to reduce intraoperative myocardial ischemia. ST segment in leads II and Vs was recorded by ST trend monitor from the time of patient's entering the operating room to the time to exit. Intraoperative myocardial ischemia occurred in 11 patients in group N and in 32 patients in group C (P < 0.0001). Prophylactic nicorandil infusion was effective to reduce intraoperative myocardial ischemia in patients with risk factors of IHD during abdominal surgery. PMID- 10689846 TI - [Sudden cardiac arrest after induction of general anesthesia: a case report]. AB - We experienced sudden cardiac arrest after induction of general anesthesia using isoflurane. The patient had had paroxysmal atrial fibrillation for one year and had been treated with digoxin and cibenzoline succinate. Sinus rhythm appeared soon after the start of closed chest compression. However cardiac arrest recurred, and we inserted a temporary pacemaker catheter to stabilize the circulatory status. She awoke from anesthesia without any complications. The diagnosis of sick sinus syndrome (SSS) was made postoperatively and she had a permanent pacemaker implanted. We thought that the hidden SSS had been the cause of this sudden cardiac arrest. PMID- 10689847 TI - [Anesthetic management for a patient with spondyloepiphyseal dysplasia congenita]. AB - An 11-year-old girl with spondyloepiphyseal dysplasia congenita in the spine was scheduled for virectomy under general anesthesia. She had slight scoliosis in the thoracic and lumbar spine, moderate funnel chest and slight thoracic kyphosis. Preoperative laboratory data were within normal range and her intelligence was normal. Her Mallanpathi's score, however, was Grade 3 and effective mandibular length/posterior depth of mandible ratio was 3.48 on the lateral view of head X ray. From these data, difficult intubation was expected. Although anesthesia was induced uneventfully using thiamylal and vecuronium, her vocal cord was not visualized under laryngoscope. Using cricoid pressure procedure, slightly left shifted vocal cord was exposed. However a 26 Fr. size endotracheal tube without cuff was too large and finally a 22 Fr. tube was inserted. During the operation patient's general condition was stable and the operation was finished without any episodes. She did not have any complications in the postoperative period. In a case of SDC, pathological changes in laryngotracheal resion should be examined and evaluated preoperatively and difficult intubation should also be always taken into account. PMID- 10689848 TI - [Tracheal mucosal bulla found on tracheal extubation in a patient with pemphigus vulgaris--a case report]. AB - A 49-year-old female with pemphigus vulgaris underwent the removal of a meningioma under general anesthesia. Neither bulla nor erosion was observed on her skin and oral cavity mucosa. She had been on prednisolone 15 mg for six years daily to avoid the recurrence of skin lesion. Anesthesia was induced and maintained with total intravenous anesthesia with propofol and fentanyl. No adverse episodes were encountered during the operative procedure. We checked the tracheal mucosa using bronchofiberscope before extubation. A small bulla was found on the tracheal mucosa, where the cuff of the tracheal tube was located. The trachea was extubated slowly under bronchofiberscopic observation, and no other bullae were found. It would have been formed by mechanical stimulation of the tracheal tube. This case suggests that we have to pay careful attention to the formation of bullae at any part of the body by mechanical stimuli during anesthetic management of patients with pemphigus vulgaris. PMID- 10689849 TI - [Anesthetic management of a patient with West syndrome]. AB - A 21-year-old female with West syndrome was scheduled for resection of hordeolum. She had an episode of convulsion at three months of age, and was diagnosed as having West syndrome at one year of age. She had epileptic seizures twice a week in spite of administration of phenytoin, clonazepam and sodium valproate. These drugs had been administered till the morning of the surgery. After premedication with atropine 0.25 mg, anesthesia was induced with propofol (12-->10-->8 mg.kg 1.h-1). The tracheal intubation was performed with vecuronium 0.1 mg.kg-1 and anesthesia was maintained with continuous infusion of propofol 6-8 mg.kg-1.h-1 and local infiltration with 1.0% lidocaine 5 ml. We administered phenytoin to prevent epileptic seizures during the surgery. No epileptic seizures occurred perioperatively. We conclude that propofol may be useful for a patient with West syndrome, and we should be careful not to lower the threshold for convulsion during the perioperative period. PMID- 10689850 TI - [Respiratory muscle weakness after prolonged use of hydrocortisone and pancuronium bromide]. AB - A 30-year-old man was admitted because of status asthmaticus. He required 7 days of artificial ventilation and was treated with hydrocortisone 1.2 g.day-1 and bronchodilaters. Pancuronium bromide 0.08 mg.kg-1.hr-1 was given for 64 hours for the ease of artificial ventilation. On day 3, severely elevated airway pressure resulted in left pneumothorax and isoflurane 1% was given for the following 2 days. Respiratory muscle weakness was evident 24 hours after discontinuation of pancuronium infusion on day 5 while full 4 twitches of TOF on the adductor pollicis muscle were seen at the time. The respiratory muscle weakness continued for another 3 days and he was extubated on day 8. Serum creatine kinase concentration rose to 2178 U.l-1 on day 6 and returned to normal on day 11. Hematurea, hyperpyrexia and metabolic acidosis were never seen during the course. Acute corticosteroid myopathy was suspected to be the cause of the prolonged respiratory muscle weakness. PMID- 10689851 TI - [A nationwide survey of anesthesia for laparoscopic and thoracoscopic surgeries]. AB - This is the report the first nationwide survey of anesthetic management for laparoscopic and thoracoscopic surgeries. We mailed a questionnaire to anesthetists of 133 hospitals in Japan and 74 completed questionnaires were returned. The number of intra-abdominal and thoracic surgical procedures has been increasing. General anesthesia was used in all cases for endoscopic surgery. The double lumen tube was selected in 79% of patients with pneumothorax for the endobronchial intubation. Patients for the laparoscopic cholecystectomy (LC) were given various types of anesthetics. Thirty-two percent of patients who underwent LC was anesthetized with inhalation anesthetics combined with epidural anesthesia for the early ambulance and postoperative pain control. The maximum length of time for LC surgery was 12.5 hr. Complications related to laparoscopic surgery included bile duct injuries in 72 patients, postoperative bleeding in 32 patients, vascular injuries in 29 patients, pneumothorax in 26 patients, bronchial intubation in 17 patients, gas embolism in 11 patients, bowel injuries in 9 patients and postoperative ileus in 7 patients. Administering anesthesia for endoscopic procedures requires precise knowledge of the surgical procedures, physiologic changes and complications of the pneumoperitoneum, and one lung ventilation. PMID- 10689852 TI - [Manual chest compressor for cardiac massage for patients transported on a litter -Part 2: Return to the air balloon method and the future direction of the study]. AB - The prior idea of sternal compression with an inflatable balloon inserted between the chest wall and the precordial metal plate was not realized due to the compressibility of gases. Based on the thoracic pump theory of cardiac massage, an accordion type balloon with a wider contact area to the chest wall was made after many years' of unsuccessful trials of various energy transmission methods. Since the compression power to the chest with this device does not exert extra weight underneath the patient, it can be used on a patient being transported on a litter. It is a simple, safe and light weight device and can be used on a helicopter as well, because the balloon can be inflated from an adjacent seat in the helicopter. Applying the principle of lever, it demands less muscle work compared with the conventional method of cardiac massage. Therefore, it may be useful even in hospitals. For the push-pull method of cardiac massage, however, an electric device may be promising when a.c. electricity or a light weight battery is available. PMID- 10689853 TI - Looking to the past for the sake of the future. PMID- 10689854 TI - From ABO to DNA... PMID- 10689855 TI - A profession in flux. PMID- 10689856 TI - The role of the medical examiner in the euthanasia notification procedure in The Netherlands. AB - The objective of the study was to provide an insight into the role of the medical examiner in the euthanasia notification procedure in The Netherlands. At the beginning of 1996 a representative group of 116 medical examiners was interviewed. The study found that there was a considerable variation in the way in which the medical examiners fulfilled their tasks. In all cases, after the physician-assisted death had taken place, the medical examiner investigated whether the attending physician had met the requirements for prudent practice, and in approximately 75% of the cases he reported his findings to the Public Prosecutor. In 78% of cases the attending physician was well known to the medical examiners, who were general practitioners, and in a third of the cases this influenced their assessment. Seventy-six per cent of the medical examiners, 61% of the members of the public prosecution and 47% of the physicians thought that it is the task to the medical examiner to review whether the requirements for prudent practice have been met by the attending physician. In conclusion further specification of the task of the medical examiner would appear to be beneficial to increase uniformity in the procedure. In particular, it should be determined whether it is the medical examiner's responsibility to review whether the requirements for prudent practice have been met. It should also be taken into consideration whether the position of the general practitioner medical examiner is sufficiently independent to make an objective report. PMID- 10689857 TI - Autopsy in elderly psychiatric inpatients: a retrospective review of autopsy findings of deceased elderly psychiatric inpatients in north Cheshire 1980-1996. AB - In this 17-year review of death certificates of elderly inpatients of a large psychiatric hospital in North Cheshire, the frequency, trend and value of performing autopsies were examined. Details of death certificates were compared with certificates issued after post-mortem examination to see whether an autopsy yielded any additional or relevant information about conditions that are not directly related to death but might well be of importance to public health. The rate of post-mortem examination, at 9.5% of total hospital deaths, did not show any significant trend over most of the review period. The vast majority of autopsies examined had been requested by the coroner and not by the clinicians. The review showed that an autopsy may be of some value in providing more information regarding any underlying causes of death in elderly psychiatric patients, but has no value in ensuring higher rate of the recording of conditions such as dementia, in particular Alzheimer's disease. Selective hospital autopsy in elderly psychiatric patients to verify, neuropathologically, the clinical diagnosis of Alzheimer's disease, will improve our diagnostic accuracy and provide valid statistics to be used in estimating prevalence, trends, risk factors and for use in all aspects of future research into Alzheimer's disease. PMID- 10689858 TI - Russian forensic psychiatry. AB - Two forensic psychiatric clinics visited on an exchange trip to Russia are described. The legal system for mentally disordered offenders in these institutions is also summarized, as is the training of psychiatrists in Russia. PMID- 10689859 TI - An exploration of the concept of loneliness in forensic psychiatry. AB - Subjective aspects of social support and loneliness were examined in a group of 30 forensic inpatients in secure units and were compared to a control group of 31 surgical inpatients. The secure-unit patients were more lonely and experienced the deficiencies in their social relationships in a different way to the controls. Loneliness for the secure-unit patients was a more unpleasant experience and more associated with difficulties in forming close attachments. There was a high correlation between loneliness and the personality trait of psychoticism in the secure-unit group which was not found in the controls. These results underline the serious problems of these patients with relationships and suggest the development of intervention programmes. PMID- 10689860 TI - Sudden death during restraint: a study to measure the effect of restraint positions on the rate of recovery from exercise. AB - A small number of mental health patients have died suddenly following violent behaviour and restraint by staff. The safety of certain restraint positions has been questioned. This study evaluates two control and restraint (C & R) positions commonly used by health service staff. A repeated measures design was used to study rate of recovery from exercise in volunteer staff, measured by pulse oximetry, comparing the restraint positions with a seated (control) position. It was found that the recovery time for pulse rate of subjects restrained in a face down position was significantly longer than for subjects restrained in a face-up position. No significant findings were made in terms of comparison between the control position and the restraint positions, and no significant changes in oxygen saturation were noted during restraint. It is concluded that restraint position may be a factor in death during restraint, but only where other factors contribute to the overall situation. PMID- 10689861 TI - Sudden unexplained death syndrome. AB - A retrospective investigation of 51 cases of sudden unexplained death syndrome (SUDS) reported to the Medico-Legal Centre in Dammam during the period January 1995 to June 1997 was carried out. The vast majority of SUDS victims were Indians (43%). The study reports SUDS in non-East Asian subjects, including indigenous Saudis, for the first time. Autopsy examination was carried out on 22 subjects and did not reveal any significant pathological lesions which could fully explain the sudden death. However, seven cases showed mild to moderate cardiac hypertrophy (of whom two had mild to moderate coronary stenosis), and another four showed a similar degree of coronary narrowing without any evidence of myocardial hypertrophy. Severe pulmonary congestion and alveolar haemorrhage were noticed in 18 of the autopsied cases. Microbiological tests were performed on different specimens from 27 subjects and showed significant bacterial growth in seven cases. The paper reviews different hypotheses explaining SUDS and suggests areas of further study in such deaths. PMID- 10689862 TI - Increasing the power of psychiatric court diversion: a new model of supra district diversion centre. AB - The object was to develop and evaluate a new concentrated model of psychiatric diversion scheme at the magistrates' court, designed to maximize the potential of such interventions. A one-year prospective study was undertaken of a consecutive series of 264 referrals to the new diversion project at an Inner London magistrates' court, with concurrent examination of police station custody records, magistrates' court returns, hospital admission data and remand prison transfer records for an area with a population of 500,000. The results showed that this one scheme originated 12.8% of all the unrestricted hospital orders in England under section 37 of the Mental Health Act 1983, 4.2% of section 35 orders, and 6.4% of section 48 and 48/49 remand prisoner transfers. Of all arrests in the central London area, 0.46% were referred to the scheme, with 0.28% being admitted. The seriousness of the charge did not have a significant effect on whether or not admission was achieved (p = 0.5365). The new model is a powerful intervention in the assessment and diversion of mentally disordered offenders. Similar supra-district diversion centres may have a role to play in other areas, complementing other local diversion exercises, some of which might better be relocated to the police station. PMID- 10689863 TI - Methadone-related deaths: data from 18 coroners' jurisdictions in England. AB - This paper examines the methadone-related deaths (MRD) among entire cases of inquest on drug-related cases in 18 coroners' jurisdictions in England which were entered on the National Programme on Substance Abuse Deaths during a six-month period in 1997. In 154 deaths methadone, either prescribed or not prescribed, was reported to be the substance directly implicated in the death of 40 individuals. The MRD between the prescribed and non-prescribed cases were compared in respect to various demographic variables and associated risks. The main findings reported are that the majority of deaths in this sample were in cases where methadone had not been prescribed (72%) and that there was a significant difference in age between the methadone prescribed (median = 22 years) and non-prescribed groups (median = 37 years) (Mann-Whitney U = 57.5, p = 0.01). Also significant differences amongst the two groups in respect of the coroners' verdicts are reported. The authors suggest more stringent controls around the prescription and dispensing of methadone, along with measures to alert the population at risk of the hazards of using methadone in a non-controlled fashion. PMID- 10689864 TI - Observation on the recent examination of bones from St David's Cathedral. AB - Bones discovered in 1866 walled up in St David's Cathedral, West Wales were thought possibly to be those of St David and his companion St Justinian, both of whom died in the late 6th or early 7th century. Examination and radio carbon dating of the bones suggested that these were not from St David nor St Justinian. Some of the bones could be the remains of St Caradoc, a 12th century hermit. It is likely, however, that the bones are remains of clergy who, for reasons not yet understood, were re-interred into the wall. PMID- 10689865 TI - Forensic medicine in the Rivers State of Nigeria: experience in four rural general hospitals. AB - A retrospective study of 375 consecutive medicolegal cases seen in four peripheral general hospitals in the Rivers State of Nigeria over a five-year period (March 1984 to February 1989) was undertaken. The most common indications for forensic medical consultation were assaults (78.6%) using clubs, sticks, fists and machetes as weapons, road traffic accidents (9.1%) and sexual offences (7.7%). The proportions of accidental deaths (4.3%), homicidal deaths (2.9%), sudden natural deaths (0.5%), maternal deaths (0.5%) and suicidal deaths (0.3%) were much lower. The male-to-female ratio was 1.4 to 1. The patients' ages ranged from 10 months to 75 years, with a mean of 31.6 years. Twenty-three cases (6.1%) were children, while the remaining 352 cases (93.9%) were adults. The study showed that for those cases which do come to the pathologist's attention, forensic personnel and laboratory services are inadequate in the peripheral parts of Nigeria. The study also highlighted the possible range of medicolegal problems of which the medical practitioner should be aware, even if he is practising in the rural non-urbanized areas of Nigeria. The study shows that not all deaths are registered in Nigeria. PMID- 10689866 TI - Maternal mortality following the use of misoprostol. AB - Three cases of maternal death following the use of misoprostol are presented. In two of the cases the drug was used to procure illegal abortion, and in the third case maternal death occurred following uterine rupture, after misoprostol was used clinically for induction of labour. A brief review of the usage of misoprostol is presented, including recommendations for its safe use. PMID- 10689868 TI - The HIV positive mother. PMID- 10689867 TI - A fatal nail gun injury--an unusual ricochet? AB - An 18-year-old construction worker suddenly collapsed while handling a power actuated nail gun and died shortly after. A neat, almost circular puncture wound was found on the front of his left chest. No fire-arm residues were detected on the surrounding skin. The police stated that it was an accidental injury, at a construction site, where a nail fired from a nail gun by the deceased had deflected off the wall and struck him on the front of the chest. Since the entry wound appeared to be a neat hole, and that too on the front of the left chest overlying the heart area, there was reluctance on the part of the pathologist to accept it as an accidental injury due to a ricochet. A visit to the scene, interrogation of witnesses, examination of the alleged tool and post-mortem X-ray of the deceased were undertaken prior to autopsy. A bent nail was found in the heart. The scene visit and the subsequent autopsy revealed that the nail took a roughly circular flightpath after it had struck the wall, all the while travelling with its pointed end directed forward. Within the body too, the nail maintained the same path. Various medicolegal issues are discussed pertaining to nail-gun injuries. The importance of a visit to the scene, examination of the alleged tool, interrogation of witnesses and the X-ray of the body, all prior to autopsy, are emphasized. The conclusion was: accidental death due to the unusual ricochet of a nail. PMID- 10689869 TI - Use of granulocyte colony-stimulating factor for treatment of aplastic anemia. AB - Over the last ten years, recombinant human granulocyte colony-stimulating factor (rh G-CSF) has been widely used in the treatment of aplastic anemia (AA). It has been shown to facilitate the recovery of neutrophil count and useful for complicated bacterial or fungal infections. However, recent randomized clinical trials showed that the addition of rh G-CSF to immunosuppressive therapy had no clinical benefit for the prophylaxis of severe infections. These results suggested that rh G-CSF should be used for the treatment of infectious complications, not for the prophylaxis of infections in patients with AA. PMID- 10689870 TI - Progress in diagnosing herpesvirus infections. AB - As molecular biology has developed, several new diagnostic techniques have found application in the clinical setting. The use of the polymerase chain reaction (PCR) assay to study the molecular biology of microbial organisms has led to unparalleled advances, largely due to the rapidity with which results can be obtained. The sensitivity and specificity of PCR detection of viral DNA for diagnostic purposes are remarkable. With such excellent sensitivity, PCR is destined to become a useful diagnostic tool in herpesvirus infections. However, it is well known that herpesviruses establish latency after primary infections and that they can often be reactivated under various conditions. Because of the high sensitivity of PCR, detection of virus sequences by this method does not necessarily imply a disease state. We must be careful not to overdiagnose conditions in a clinical setting based on a PCR assay. PMID- 10689871 TI - NK and NK-related neoplasms. AB - Owing to the immunological progress in recent years, differentiation and maturation process of NK cells or NK-related cells has been made clear. Also, their neoplastic counterparts, namely NK cell related lymphoma/leukemia have been characterized. In this paper, we will report NK cell neoplasms based on the new classification of malignant lymphomas. Among these, nasal lymphoma is one of the most clearly defined entities, followed by NK cell leukemia/lymphoma, and blastic/blastoid NK cell leukemia/lymphoma. PMID- 10689872 TI - A new operation for noncorrectable biliary atresia. AB - An improved operative technique to transect the fibrous cord by dividing the ligamentum venosum (Arantius' canal) is described for noncorrectable biliary atresia. The Arantius' canal is situated cranial and posterior side to the bifurcation of the umbilical portion and the portal branch of the Quinous' segment 3. The portal vein is fully mobile and the porta hepatis can be widely exposed by dividing the Arantius' canal. The fibrous cord of the porta hepatis can easily be dissected posteriorly and laterally where there is an extensive number of bile ducts. Eight patients with biliary atresia underwent this procedure. Jaundice resolved completely (serum total bilirubin concentration: < or = 1 mg/dl) in 7 patients within 40 days. Postoperative cholangitis did not occur. By dividing the Arantius' canal, the portal vein comes free from the portal fissure to make the hepatic hilum wider, and surgeons are able to work within a larger porta hepatic space without causing portal vein compression. Free drainage of the bile from the porta hepatis may prevent postoperative cholangitis and promote resolution of jaundice. PMID- 10689873 TI - Effects of sodium hyaluronate on experimental osteoarthritis in rabbit knee joints. AB - The aim of this study was to examine the effects of intraarticular administration of hyaluronan (HA) on cartilage degradation. Using a partial menisectomy model of osteoarthritis (OA) in the rabbit knee, the authors investigated the catabolic and anabolic changes induced by intraarticular injection of HA. To analyze anabolic changes, the authors assessed cell proliferation by measuring [3H] thymidine uptake, and proteoglycan biosynthesis by noting [35S] sulfate incorporation. For catabolic changes, messenger ribonucleic acid (mRNA) expression of interstitial collagenase (MMP-1), stromelysin-1 (MMP-3), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in cartilage and synovium were detected with reverse transcriptase polymerase chain reaction (RT-PCR). Of significance for blocking the development of early OA in chondrocytes was the finding that total proteoglycan synthesis in the HA treatment group was significantly higher than in the controls. At the mRNA level in cartilage and synovium, HA inhibited MMP-3 and TIMP-1 production in the same way in the HA treatment group, while not affecting MMP-1 production. Thus it can be concluded that HA affects cartilage catabolism and anabolism to prevent the progress of OA. PMID- 10689874 TI - [Empiric and mechanistic evaluation model for lactate concentration time curves in endurance tests]. PMID- 10689875 TI - Acute myeloid leukemia in the elderly:--159 Nagoya case studies--Nagoya Cooperative Study Group for Elderly Leukemia. AB - To obtain background information on elderly acute myeloid leukemia (AML), unselected data covering 159 patients aged 60 years or over with AML from 14 hospitals in Nagoya, Japan was analyzed retrospectively. Among these patients, 119 had de novo acute AML, 32 had AML which evolved from myelodysplastic syndrome (MDS-AML), and 8 had other types of leukemia. The survey showed that MDS-AML tended to be more prevalent in patients aged 70 years and older and that MDS-AML showed a significantly more severe degree of leukopenia and anemia than de novo AML. MDS-AML also showed a significantly lower complete remission (CR) rate than that of de novo AML [6.9% (2/29) vs 58.3% (67/11), P < 0.01] and significantly shorter survival times than those of de novo AML [median: 3.6 months vs 9.6 months, P < 0.01 (generalized Wilcoxon test; GW]. In de novo AML, the proportion of patients treated with conventional therapy (CT group) decreased significantly, and that of those with attenuated therapy (AT group) increased significantly as age elevated (P < 0.01). The CT group showed a significantly higher CR rate (65.4% vs 41.2%, P < 0.05) and a significantly longer survival period than those of the AT group [median: 11.6 months vs 4.8 months, P < 0.05 (GW)]. Overall survival rates of the older age groups became significantly shorter with aging [P < 0.01 (GW)]. PMID- 10689876 TI - [The result of mass screening of 1997 for prostatic cancer in Isesaki City]. AB - PURPOSE: Screening by only prostate specific antigen (PSA) for prostate cancer was started since PSA had been added to mass screening as one of check lists in 1997 in Isesaki city, Gunma pref. We expected PSA screening to be introduced into other areas. We therefore studied how to perform a screening procedure for prostate cancer as well as discussed our result of the screening conducted lately. MATERIALS AND METHODS: 1,382 out of 1,423 Isesaki citizens who took mass screening aged 40 to 64 were chosen. Regardless of age, men with a serum PSA level equal or larger than 4.1 ng/ml (Tandem R) were selected for second screening since we determined it was a cut-off level for further check-up. Of those men, 38 were requested for second screening and actually only 24 took it. All these men took PSA check-up again, furthermore 23 took transrectal examination (TRE) and/or transrectal ultra sonography (TRUS) except for one of them. The next screening was requested for sixteen of them. Prostate biopsy was conducted for all of them. RESULTS: More old men took screening and were diagnosed prostate cancer. The findings derived from such diagnosis showed one of them aged 50 to 59 and six of them aged 60 to 64 had the cancer. Moreover, four out of twenty with PSA level ranging 4.1 to 10.0 ng/ml and all of three with PSA level over 20.0 ng/ml had the cancer. Five out of sixteen with a positive sign for further PSA check-ups had the cancer. All the three suspect of the cancer by TURS and DRE had prostate cancer. Two of seven with PSA negative showed suspicion of prostate cancer and had the cancer. No neo-adjuvant and total prostatectomy was conducted for four with 4.0 to 10.0ng/ml diagnosed T2N0 M0. One of them with PSA equal or over 20.0 ng/ml was diagnosed T3N0M0. After hormone therapy its PSA decreased to that equal or under 0.5 ng/ml. Total prostatectomy was conducted for it. CONCLUSION: It is not proved that only PSA mass screening for prostatic cancer contributes to detect early cancer and better prognosis cure case. For the proof, it will be nessary that PSA mass screening is examined more people in the wide area. We conclude men aged 65 to 69 also should take PSA check-up based on epidemiological feature of prostatic cancer. PMID- 10689877 TI - [Autologous blood transfusion in surgery of urologic malignant tumor]. AB - BACKGROUND: Autologous blood transfusion has been widely endorsed, because of the adverse effects attributed to homologous blood transfusion. So we employed autologous blood transfusion to avoid homologous blood transfusion in operation of urological malignant patients. We reviewed our experience with autologous blood transfusion in 48 patients. METHODS: A total of 48 patients underwent operation with 400 to 1,200 ml preoperative autologous blood donation, in 41 patients with administration of erythropoietin and 7 patients without erythropoietin. The details of operations are radical nephrectomy in 18 cases (2 cases were bilateral), radical nephro-ureterectomy in 2 cases, retroperitoneal lymph node dissection (RPLND) in 2 cases, radical prostatectomy in 12 cases and radical cystectomy in 14 cases. RESULTS: The volume of surgical blood loss were 381 +/- 522 ml in nephrectomy (1,158 +/- 202 ml in bilateral case), 517 +/- 5 ml in radical nephro-ureterectomy 636 +/- 574 ml in RPLND, 665 +/- 291 ml in radical prostatectomy and 1,123 +/- 417 ml in radical cystectomy. Only three cases needed homologous blood transfusion. CONCLUSION: We can avoid homologous blood transfusion in 94% of patients. Autologous blood transfusion is recommended as safe and convenient. PMID- 10689878 TI - [Kinetics of peripheral blood CD34-positive cells and the optimum timing for harvesting peripheral blood stem cells during BEP chemotherapy in patients with testicular germ cell tumor]. AB - PURPOSE: Recently, high-dose chemotherapy with peripheral blood stem cell (PBSC) rescue has been developed for poor risk testicular germ cell cancer. In this study, we investigated the optimum timing for harvesting PBSCs with the use of bleomycin + etoposide + cisplatin (BEP) chemotherapy, which is a well known first line regimen for the testicular cancer. MATERIAL AND METHOD: Peripheral blood CD34-positive cell ratios were measured during a total of 10 courses of BEP chemotherapy in 6 patients with metastatic germ cell cancer between 1996 and 1998. We performed 4 apheresis in 3 patients during this period. Recombinant human granulocyte-colony stimulating factor (rhG-CSF) was administrated from the day on which the neutrophil count decreased less than 1,000/microliter. RESULTS: The peripheral blood CD34-positive cell ratios became maximum (3.0-24.6%; average 10.0%) on the day 18 to 21 (median day 19) of BEP chemotherapy with rhG-CSF administration. The maximum ratios of peripheral blood CD34 positive cells were achieved when the number of leukocyte were 6,880-23,600/microliter and exceeded 6,000/microliter after the 18th day of BEP chemotherapy. The average number of collected CD34 positive cells was 9.5 x 10(6)/kg at a single apheresis, and 12.6 x 10(6)/kg per patient. CONCLUSION: Efficient hematopoietic progenitor cells were mobilized by BEP chemotherapy with rhG-CSF administration of first-line setting. Our results suggest that the optimum timing of PBSCs harvest is the day when the numbers of leukocyte exceed 6,000/microliter after the 18th day of BEP chemotherapy and the following day. PMID- 10689879 TI - [The quality of life after radical prostatectomy measured by general health questionnaire and visual analogue scales]. AB - OBJECTIVES: The impact of radical prostatectomy on the quality of life (QOL) of patients were evaluated. MATERIALS AND METHODS: A total of 22 patients who underwent radical prostatectomy for clinically localized prostate cancer entered this study. Patients were asked to complete a questionnaire containing the general health questionnaire (GHQ) and a series of questions evaluating voiding function, incontinence and sexual dysfunction before and after the operation. In addition, the visual analogue scaled (VAS) questionnaire containing incontinence and sexual dysfunction was applied. RESULTS: No significant differences in GHQ were found between pre- and post operative status, but disease-targeted QOL such as sexual function was affected after the radical prostatectomy. In the points of incontinence and sexual dysfunction, VAS questionnaire significantly correlated with those of categorical questionnaires. CONCLUSIONS: There results suggest that GHQ is not affected, but disease-targeted QOL in some categories of sexual function is affected by radical prostatectomy, and that VAS questionnaires are not only useful for assessing the disease-targeted QOL but also easy to quantify QOL of the patients. PMID- 10689880 TI - [Suicidal attempts in three postoperative patients with renal cancer after alpha interferon withdrawal]. AB - We report 3 cases of suicide attempts in postoperative patients with renal cancer after alpha interferon withdrawal. In the first patient, depression occurred during interferon therapy, and remained after interferon withdrawal. A suicide attempt occurred 7 months after interferon withdrawal. In the second and third patients, depression did not occur during interferon therapy, but suicide attempts occurred 40 days and 7 months after interferon withdrawal, respectively. Depression does not always disappear after interferon is discontinued. Psychiatric supervision should be continued even more frequently after interferon withdrawal. The increased risk of psychiatric side effects due to interferon, as well as their severity, suggest that interferon should be administered with caution. PMID- 10689881 TI - [Familial isolated hyperparathyroidism: a report of two cases]. AB - A family (a brother and a sister) of the familidal isolated hyperparathyroidism (FIH) was reported. The older brother with age of 58 year-old was pointed out hypercalcemic while examining his hypertension and proteinuria. He had high levels of serum total and ionized calcium, intact-PTH and gastrin, and hypophosphatemia. His neck CT scan revealed swelling of the two parathyroid glands in each side. He underwent resection of the tumors and the auto implantation of the glands under diagnosis of primary hyper parathyroidism. Histopathology was diagnosed to be hyperplasia of the parathyroid glands. The younger sister with age of 52 year-old was referred to our clinic because she was suffering from recurrent urolithiasis. Biochemical examination of her blood sampling resulted in very resemble values of her brother mentioned above. Her neck CT scan showed three tumors consisting of each one at the bilateral parathyroid glands and one in the thymic region. She underwent resection of the tumors and the auto-implantation of the glands and histopathological diagnosis was hyperplasia as same as her brother's one. The postoperative courses of these cases have been uneventful for four years. FIH is a low significant disease of which ten lineages have been reported in Japanese literature although it should be differentiate with such a disease of multiple endocrine neoplasms. PMID- 10689882 TI - [Renomedullary interstitial cell tumor: a case report]. AB - Renomedullary interstitial tumor is a common tumor in the renal medulla, present in 26-41% of consecutive autopsy specimens. However clinically evident case is infrequent because this lesion is usually small (less than 3 mm). We report a case of renomedullary interstitial tumor in a 76-year-old woman. Ultrasonogram incidentally revealed a mass in the left kidney while she visited to the hospital for hypertension and unstable angina. A CT scan showed a 2 cm mass that was not clearly enhanced. MR images showed low signal intensity in both T1 and T2 images. Arteriography demonstrated no neo-vascularity. Those findings showed that this lesion was benign one or hypovascular carcinoma. So Left nephrectomy was performed and histological examination revealed a renomedullary interstitial tumor. PMID- 10689883 TI - [Diagnostic radiology in acute pediatric abdomen]. AB - Acute pediatric abdomen is a very common clinical problem. Clinical and laboratory findings, however, are nonspecific or confusing in many instances. This review article focuses on strategy in diagnosing acute pediatric abdomen. A variety of diseases such as appendicitis, gastroenteritis, mesenteric adenitis, intestinal intussusception, Henoch-Scholein purpura, Crohn's disease, Meckel's diverticulitis, duodenal ulcer, congenital biliary dilatation, ovarian torsion, and anomaly of the internal genitalia are discussed in this article. Selection of an appropriate imaging modality is essential to ensure prompt management. In the majority of cases, ultrasound can provide specific diagnoses, whereas in others valuable supplemental information can be obtained. CT will be reserved for selected patients in whom further information is needed. Indications of MR imaging in the management of acute pediatric abdomen are currently limited. MR imaging, however, is indicated on an emergency or semi-emergency basis in selected conditions including anomaly of the internal genitalia, ovarian torsion, and congenital biliary dilatation. PMID- 10689884 TI - Initial clinical application of cone-beam CT scan in pulmonary imaging. PMID- 10689885 TI - [Three-dimensional imaging of hepatic and intrahepatic portal veins with helical CT: determination of optimal volume of contrast medium by intravenous injection using MIP technique]. AB - In this study, the optimal volume of contrast medium in the liver for three dimensional (3D) imaging of the hepatic and portal veins by helical CT were determined by intravenous injection using the MIP technique. In the 48 cases examined, CT images of the liver were obtained following the administration of contrast medium (90, 120, or 150 ml and 1.0 <, < or = 1.5 ml; 1.5 <, < or = 2.0 ml; 2.0 <, < or = 2.5 ml or 2.5 <, < or = 3.0 ml/kg) for determination of the optimal volume. The mean body weight of the patients was 59 kg. Contrast medium (Iopamidol 300 mgl/ml) was injected at a rate of 3 ml/sec, and scanning was initiated 70 sec after the beginning of injection. Images were obtained throughout the entire liver using 5-mm collimation. MIP images were graded from poor to excellent based on their visualization of the hepatic vessels. Images produced with 120 ml of contrast medium presented excellent images of hepatic vessels, superior to those produced with 90 ml (hepatic vein: p < .001, portal vein: p < .001). Images produced with 2.0 <, < or = 2.5 ml/kg of contrast medium presented excellent images of the portal vein, superior to those produced with 1.5 <, < or = 2.0 ml/kg ml (p < 0.05). It is evident from the present data that a contrast medium volume of more than 120 ml or 2.0 <, < or = 2.5 ml/kg is sufficient for three-dimensional imaging of hepatic vessels. These images may be a useful diagnostic tool in patients with hepatic disease. PMID- 10689886 TI - [Percutaneous mechanical thrombectomy for thrombosed vessels with a hydrolyser (hydrodynamic thrombectomy catheter): clinical experience]. AB - A hydrodynamic thrombectomy catheter was prospectively evaluated for the treatment of thrombosed vessels. Seven patients (7 males: age range from 56 to 82 years; mean age: 79 years) presenting with acute or chronic occlusion of peripheral native arteries (n = 6) and dialysis shunt (n = 1) were treated with the hydrolyser (Cordis, Johnson and Johnson, Japan). Mean occlusion time was 135 days (range: 2-300 days), and mean thrombus length 16 cm (range: 5-20 cm). Removal of the thrombus was successful in five patients (71%), regardless of the length of the thrombus. Mean procedure time was 20 minutes (range: 15-30 minutes). No major complications occurred. Adjunctive thrombolysis was required for persistence of the residual thrombosed distal vessel in one patient. Adjunctive balloon angioplasty was performed in two patients (one native vessel and one dialysis shunt), and stent placement was performed in one patient (dialysis shunt). In two unsuccessful cases, the hydrolyser could not be advanced to the distal side because of the solid thrombus. Therefore, thrombolytic therapy was chosen. However, this therapy failed because the guidewire did not pass within the thrombus. Surgery was performed in these two patients. We conclude from this clinical experience that percutaneous thrombectomy with a hydrolyser is a promising technique for the treatment of thrombosed vessels. Especially in the acute stage of thrombosed occlusion, percutaneous thrombectomy with a hydrolyser is superior to thrombectomy with a Fogarty balloon catheter because of its shorter procedure time and fewer complications. PMID- 10689887 TI - [Usefulness of selective cerebral angiography by transradial approach]. AB - Transradial angiography has recently emerged as an alternative to the transfemoral or transbrachial approach, especially for coronary procedures. However, there have been few studies on cerebral angiography using the transradial approach. The purpose of this study was to assess the outcomes, complications, and limitations of selective cerebral angiography via the transradial approach. Selective cerebral angiography by the right transradial approach using 100-cm-long 4-F catheters was performed in 83 patients. Using five types of catheters, the success rates of selective catheterization to the right vertebral artery, right common carotid artery, left common carotid artery, and left vertebral artery were 40/44 (91%), 68/68 (100%), 62/62 (100%), and 14/25 (56%), respectively. Puncture failed in one patient, and a guidewire could not be introduced beyond the radial artery loop in one patient. Radial artery spasm occurred in one patient, but was relieved immediately after nitroglycerin injection through the sheath with side holes. Subcutaneous bleeding occurred in six patients, but no obvious hematomas were noted. Occlusion or stenosis of the radial artery occurred in five patients, but no ischemic symptoms were observed in any of the cases. This study suggested that selective cerebral angiography can be performed safely using the transradial approach. PMID- 10689888 TI - [Assessment of non-ionic contrast agents in reducing the risk of side effects: analysis on the basis of voluntary willingness-to-pay measured by the contingent valuation method]. AB - The benefit of replacing ionic contrast agents with non-ionic ones was assessed by employing cost-benefit analysis, a method of medical economic analysis. The benefit derived from replacing ionic with non-ionic contrast agents was assessed by a questionnaire survey of patients using the willingness-to-pay method based on the contingent valuation method. This questionnaire survey was conducted on 204 patients in Shimane Medical University Hospital during the period from October to December 1998. The result of analysis showed that when ionic contrast agents are replaced with non-ionic ones, patients' willingness-to-pay stands at a median value of 12,500 yen and a mean value of 17,082 +/- 1,049 yen. These figures are identical with or larger than the NHI-price differences (12,266 14,234 yen; average 13,287 yen), suggesting that patients think the benefit of reduced side effects from non-ionic contrast agents has a value that is equal to or higher than the actual NHI-prices of these agents. Further, analysis of patient profiles indicated that patients' willingness-to-pay went up with age and income but decreased when age exceeded 60 years, a finding which also suggests that the willingness-to-pay amount is closely related to the economic strength of patients. PMID- 10689889 TI - [Preliminary study of calculating cerebral arterial blood oxygen saturation using MRI]. AB - To assess whether cerebral arterial blood oxygen saturation (SaO2) can be calculated by EPI, we examined the relationship between peripheral SaO2 and T2+ signal intensity (SI) changes in the brain in three normal subjects, using 1.5 Tesla MRI. To decrease SaO2, hypoxia was induced by 100% helium-gas inhalation (60 sec). SI declined as SaO2 decreased during helium inhalation, while rapid recovery of SI to the baseline was noted with recovery from hypoxia. The differential effective transverse relaxation rate was closely correlated with SaO2 (r > 0.94). Consequently, using MRI, we were able to calculate arterial SaO2. PMID- 10689890 TI - [Evaluation of in vitro human blood transparentized using near-infrared light]. AB - We developed a new technique for the in vitro imaging of transparent human blood and examined the resolution of a test chart in transparent blood. We utilized mainly three devices: a laser diode (wavelength 833 nm) that served as the light source, a near-infrared camera, and a fiberscope adapted for the camera. Blood was collected from a human femoral artery. We observed the images of transparent blood and the fiberscope image of a target in the blood using the light. These results indicate that further improvements in this system can be expected and real-time viewing by angioscope may be realized in vivo. PMID- 10689891 TI - [Development of DICOM image browser for Macintosh computer]. AB - Although many medical image browsers are available today, most are extremely expensive. To solve this problem, we developed a Digital Imaging and Communications in Medicine (DICOM) image browser (Medical Image Browser) for Macintosh computers. A comparison between this software, Advantage Workstation, NIH Image, and Graphic Converter suggested that this software is useful for diagnosis on the Macintosh desktop. This software is available at wwwl.odn.ne.jp/cak42860. PMID- 10689892 TI - [Pneumonia in the older people]. PMID- 10689893 TI - [Insulin receptor and aging]. AB - The potential link between aging and insulin signaling has attracted substantial attention since several decades ago, on the basis of evidence including age related increase in incidence of insulin resistance, insulin resistance and type 2 diabetes in accelerated aging syndromes and lifespan extension by caloric restriction in rodents. In addition, the intensive investigations in C. elegans in the 1990's, which have identified insulin signaling components including daf 2, age-1 and daf-16 as the genes whose mutations lead to lifespan extension, shed new light on molecular mechanisms underlying aging. As suggested by the genetic studies in C. elegans, it was recently demonstrated that FKHR, FKHRL1 and AFX, which are mammalian homologues of daf-16 forkhead transcription factor, function downstream of insulin signaling and Akt/PKB under cellular conditions. However, it is an open question whether insulin signaling components, including forkhead transcription factors, play a critical role in aging and longevity in mammals as well as in C. elegans. Increasing evidence concerning C. elegans indicates that augmented resistance to stress, in particular, that to oxidative stress is involved in lifespan extension by genetic mutations of insulin signaling components. The intriguing finding that signals from the reproductive system regulate lifespan by modulating the activities of insulin signal transduction pathway in C. elegans suggests a possibility of co-evolution of reproduction and aging. The significance of studies on C. elegans with regard to human aging is discussed. PMID- 10689895 TI - [Palliative care]. PMID- 10689894 TI - [Neurological diseases in the elderly]. PMID- 10689896 TI - [Diabetes mellitus in the elderly]. PMID- 10689897 TI - [Happiness and background factors in community-dwelling older persons]. AB - In order to maintain and improve mental health of elderly people living in the community, a cross sectional survey was conducted to elucidate their happiness and background factors. The subjects were elderly persons living in the community who were able to fill in the questionnaire themselves. The study employed the self-recording questionnaire forms used in Kahoku Longitudinal Aging Study by Matsubayashi et al. Happiness was assessed using a visual analogue scale. Out of 2,379 elderly persons who were able to fill in the questionnaire by themselves in 2,361 (99.2%) returned the questionnaire sheets. After removing inadequate responses, analysis was possible for 1,873 (78.7%) (860 men (average age 72.7 years) and 1,013 women (average age 72.8 years). Among those with greater happiness, the ratio of those living with others (p = 0.0051) was high as well as those with spouses (p = 0.0240), without a history of hypertension (p = 0.0096) and apoplexy (p = 0.0039), not receiving medication regularly (p = 0.0039), with regular habit of walking (p < 0.001), or with work (p < 0.001). As for the relationship between happiness and various scores, the higher the happiness scale became, the scores for ADL, information-related function, functional and emotional support network, healthy condition, appetite condition, sleep condition, memory condition, family relationships, friendship, economic condition became significantly higher (p < 0.001, respectively). On multiple regression analysis using the background factors for happiness as explanatory variables, factors such as functional support network (p < 0.001), emotional support network (p = 0.0254), healthy condition (p < 0.001), good memory condition (p = 0.0027), friendship (p < 0.001), good economic condition (p < 0.001) were significant independent contributing factors. As for the relation between SDS and happiness, the more serious the SDS score (higher score) became, the scores for the feeling of happiness became significantly smaller (p < 0.001). For amelioration of the happiness of elderly persons living in the community, attempts should be made to improve the background factors clarified by the present study by efficiently utilizing health, medical and welfare services. PMID- 10689898 TI - [Sex differences in subjective well-being and related factors in elderly people in the community aged 75 and over]. AB - Sex differences in factors related to subjective well-being were evaluated in people in their late old age by interviewing individuals aged 75 years and over living in 2 regions of Enzan City, Yamanashi Prefecture. The 17-item revised "Philadelphia Geriatric Center (PGC) Morale Scale" was used for evaluation of subjective well-being. Factors related to family status, employment, health related factors, activities of daily living, and results of physical examinations were each classified into two or more categories, and PGC Morale Scale points were compared among the categories for all subjects and separately for males and females using the Mann-Whitney test and the Kruskal-Wallis test. 1) Although the mean age of males (80.0 +/- 4.4 years) and females (80.4 +/- 4.3 years) was no significantly difference, PGC Morale Scale points were significantly higher in males than in females (p < 0.05), indicating a sex difference in subjective well being. 2) In elderly females, subjective well-being was related to many factors, and there was a particularly strong relationship between subjective well-being and both health related factors and activities of daily living. 3) In elderly males, the factors related to subjective well-being were fewer than in females, consisting only of "social opportunities", "hobbies", and "grip strength". Since factors related to subjective well-being differ between the sexes, these differences must be taken into account when evaluating subjective QOL of the elderly. PMID- 10689899 TI - [Effect of age, gender, and body fat distribution on serum leptin concentrations]. AB - The aim of this study was to clarify the relationship between serum leptin concentration and age, gender and body fat distribution. Serum leptin concentrations were determined 267 subjects (138 men and 129 women), aged 30 to 91. The thicknesses of the preperitoneal fat layer (Pmax) and subcutaneous fat layer (Smin) in the abdomen were measured by ultrasonography. Fat mass and percent fat were measured by the bioelectrical impedance analysis method. Women had higher leptin and leptin/fat mass values than men in all BMI groups (BMI < 20, 20-23.9, 24-25.9, > or = 26). The leptin concentration correlated significantly with BMI, fat mass, percent fat, waist, hip, waist/hip ratio (W/H), Pmax, Smin and serum Cr in both men and women. The leptin concentration correlated significantly with age in men only and P/S in female only. Leptin/fat mass values significantly correlated with age, fat mass, and percent fat, but not with BMI, waist, hip, W/H, Pmax, Smin and P/S in men. In women, leptin/fat mass values significantly correlated with BMI, fat mass, percent fat, waist, hip, Pmax, Smin and P/S, but not with age or W/H. Multiple regression analysis showed that fat mass and serum creatinine were significant determinants of the leptin and leptin/fat mass values both in men and women, but that age was a significant determinant of these values only in men. These results suggest that the influence of aging on serum leptin concentration exists only in men. PMID- 10689900 TI - [A case of elderly-onset systemic lupus erythematosus (SLE) complicated with severe liver dysfunction and pancytopenia due to myelofibrosis]. AB - A 67-year-old woman was admitted to our hospital with a fever. She had been experiencing arthralgia for about one month. On admission, she had a fever of 38.5 degrees C, was anemic and was experiencing tenderness in the joints of both hands, elbows and feet. Laboratory data revealed proteinuria, urinary cylinders, pancytopenia (WBC 900/mm3, Hb 9.5 g/dl, Plt 7.8 x 10(4)/mm3), liver dysfunction (GOT 414 IU/l, GPT 140 IU/l), and hyper-gamma globulinemia. Antibiotics and granulocyte-colony stimulating factor were administered intravenously. Bone marrow aspiration was unsuccessful, but a bone marrow biopsy revealed bone marrow fibrosis. Immunological examinations were positive for antinuclear antibodies, anti-deoxyribonucleic acid (DNA) antibodies, anti-double stranded anti- DNA antibodies, as well as a decreased level of serum complement and an increased level of serum immune complexes. Tests for viral antigens and antibodies known to cause hepatitis were negative. Based on these findings, a diagnosis of SLE accompanied by liver dysfunction and bone marrow fibrosis was made. Steroid pulse therapy was initiated, but her liver function deteriorated on the first day of steroid therapy, and she died three days later. SLE accompanied by myelofibrosis is extremely rare, and only 17 and cases have been reported to date. Among these reports, the present case is the second oldest subject and the first SLE patient to suffer from both myelofibrosis and severe liver dysfunction. PMID- 10689901 TI - [Interstitial pneumonitis associated with Sweet's syndrome in the elderly]. AB - Sweet's syndrome occurring during the course of interstitial pneumonitis in a 70 year-old woman was encountered. She was admitted because of dyspnea on exercise, dry cough and interstitial shadow on chest x-ray. Lung biopsy, together with other findings confirmed interstitial pneumonitis. Five days after admission, genital ulcer and aphtha on the oral mucosa were detected and exudative erythema appeared on her right shoulder, chest and back. Histological examination of the skin lesions demonstrated numerous nutrophilic infiltration in the upper dermis, indicating Sweet's syndrome. The skin eruption rapidly disappeared on treatment with colchicine. Although six months after admission interstitial pneumonitis caused respiratory failure, treatment with prednisolone and cyclophosphamide was effective. Serological and immunological tests demonstrated hyper gammaglobulinemia and positive reaction for anti SS-A antibody. Pathological examination of the lip revealed numerous lymphocyte infiltrates around the duct of the minor salivary gland, suggesting Sjogren's syndrome as the background disease of Sweet's syndrome and interstitial pneumonitis. This evidence indicating that even in elderly patients, skin lesions of Sweet's syndrome may reveal the background disease. PMID- 10689902 TI - [An extremely elderly patient with choledocholithiasis and many complications]. AB - It was very difficult to treat a 90-year-old woman for choledocholithiasis with acute obstructive suppurative cholangitis, gallbladder perforation, and a pool of bile in the right perirenal spase. Extracorporeal shockwave lithotripsy (ESWL) was performed after emergency percutaneous transhepatic biliary drainage (PTBD), but we could not perform lithotripsy successfully because of large and hard stones. Although Endoscopic sphincterotomy (EST) was performed using an ultratome by rendezvous method. Lithotripsy was finally successful, after three times endoscopic mechanical lithotripsy (EML) and procedure using an endotriptor for basket impaction. It is very important in advanced aged patients that endoscopic treatment should be performed step by step. PMID- 10689903 TI - [A case of medial medullary infarction without Dejerine syndrome]. AB - A 67-year-old man with right hemiparesis and dysarthria was admitted with right hemiparesis involving the face, hyperpathia, numbness and pain of the right body and limb except the face, and had hyperreflexia and pathological reflex in the right limb. Brain MRI on the day after admission disclosed no lesion which might explain the symptoms. Short latency somatosensory evoked potential showed a low amplitude after P14 when the right side was stimulated. Cerebral angiography revealed occlusion of the left vertebral artery. Brain MRI on the 18th hospital day disclosed left medial medullary infarction, so we diagnosed medial medullary syndrome. This case was hard to diagnose, because of the atypical features and the absence of an abnormal lesion on the initial MRI. PMID- 10689904 TI - [Otolaryngological Clinic at the Medical Academy in Poznan in 1979-1999]. PMID- 10689905 TI - [Neck reoperations in patients with laryngeal, lingual and tonsillar neoplasms]. AB - One of the most common reasons of failure in head and neck cancer treatment are metastases in regional lymph nodes. Preoperative assessment of the neck lymphatic system is an important task, but even more crucial goal is to monitor the patients after initial operation or combined treatment. The frequency of nodal recurrences ranged from 9% to 23%. It constitute nearly 50% of all treatment failures in larynx, tongue and tonsil malignancies. There are different methods of therapy in nodal recurrence treatment, but the most recommended, easily accessible and widely used is surgery. The group of 2134 patients (1580--larynx cancer, 286--tongue and floor of mouth cancer, 268--tonsil cancer) treated between 1987-1997 were operated in ENT Dept. of K. Marcinkowski University of Medical Sciences in Poznan. In 269 patients were detected the nodal recurrence. In 149 cases the recurrence was homolateral, in 48 heterolateral, in 71 patients the neck was not previously treated. 152 patients were irradiated on the neck fields just after primary surgery. The rate of recurrences was 12.6%. Authors analysed and compared the percentage of recurrences for different primary lesions: larynx, tongue and tonsil neoplasms. The period of time from the last control examination to the moment of recurrence detection, its frequency on particular nodal levels, number of recurrences and trends between years 1992-1997 were assessed. 208 patients who developed nodal recurrence had surgical salvage, 59 patients were not qualified to surgery because of the lesion extension. Neck re-operations were divided into 3 main types: selective, modified radical and radical neck dissection. The frequency of particular neck dissection types, curative rates and difficulties of performing the salvage surgery were discussed. PMID- 10689906 TI - [20-year-long experience in the large-scale fronto-lateral laryngectomy with a simultaneous reconstruction by muco-cartilagenous nasal septum flap: results of 460 cases]. AB - The late results of the larynx reconstruction by means of the mucoseptal nasal graft after an enlarged fronto-lateral laryngectomy were described. The analysis of 460 patients showed a satisfying larynx lumen in almost all cases. In 21 cases due to the neoplasm recurrence a total laryngectomy were performed. The voice after surgery was socially good, and the wide larynx lumen was obtained. PMID- 10689907 TI - [Chromosome damage in the course of laryngeal squamous cell carcinoma]. AB - The aim of the article is a review of own cytogenic studies on laryngeal cancer confronted with the literature data. Spontaneous and bleomycin-induced chromosome instability was analysed in peripheral blood lymphocytes in relation to genetic risk of cancer incidence and progression. Comparative genome hybridization (CGH) was applied to demonstrate gains and losses of DNA copy number in tumour and non tumour laryngeal mucosa. The profiles of imbalances of DNA copy number were shown to differ between metastazing and non-metastazing tumours. Preliminary data indicate a frequent loss of Y chromosome in tumour cells. The loss of heterozygosity at chromosome p53 locus (17p) has been shown to be more frequent than at chromosome locus coding 16 gene (9p). Altogether, the experiments have proven that a dynamics of chromosome aberrations is highest at the stage of metastasis. PMID- 10689908 TI - [Adenoid cystic carcinomas of the head and neck (cylindromas)]. AB - In the years 1958-1997 111 patients with adenoid cystic carcinoma in the head and neck of different localization were treated at the Department of Otolaryngology- Head and Neck Surgery. Majority of the tumors originated of the major salivary glands, mainly in the parotid gland (47 cases). The most common location among the tumor of the minor salivary glands was the palate (14 persons). The tumors arising from other mucous glands, most frequently has located in the maxillo ethmoideal complex (20 cases). The authors stress diagnostic difficulties, the necessity of a proper selection of the surgical method and postoperative radiotherapy, describing the treatment results in the patients. Radial surgical procedure of the adenoid cyctic carcinoma yelds optimal local control and survival rates. PMID- 10689909 TI - [Pleomorphic adenomas in the materials of the Department of Otolaryngology of the Academy of Medical Sciences in Poznan]. AB - In the years 1958-1997 were surgically treated in the Department of Otolaryngology Karol Marcinkowski University of Medical Sciences in Poznan 365 patients with pleomorphic adenomas. Majority of the tumors originated of the major salivary glands was parotid gland (273 cases). The given materials confirms the viewpoint that enucleation in the pleomorphic adenomas of the parotid salivary gland--based on total removal of the tumor together with the capsule and pouch constitute a completely sure surgical procedure. PMID- 10689910 TI - [Diagnostic difficulties in cases of laryngeal cancer]. AB - The span of time between the appearance of the first symptoms of larynx cancer and the moment it is diagnosed in Poland amounts to approximately 6 months. It is the result primarily of the fact that patients tend to consult a doctor late after the symptoms show as they underestimate them due to their lack of knowledge concerning their nature. In some cases the diagnosis may be difficult to give even for a specialist. The paper presents the cases of three patients with an unusual course of larynx cancer. PMID- 10689911 TI - [Otitis media with effusion]. AB - The authors present monographic data as regards causes, epidemiology, patomechanism and treatment of otitis media with effusion. The attention is paid to connections between this pathology and acute, recurrent otitis media and chronic eustachian tube insufficiency. The authors underline occult onset, asymptomatic and chronic character of this disease, irreversible consequences and hearing impairment. Among treatment methods the most effective seemed to be the surgery one: adenotomy with simultaneous tympanostomy. The authors give the schedule of management of otitis media with effusion and indications to ventilation tube insertion. PMID- 10689912 TI - [Critical evaluation of the audiological test results in children]. AB - Factors influencing results of objective hearing examination in children, i.e. auditory evoked potentials, based on own experiences are presented. "Cross-check" principle as well as observation of dynamic changes concerning development of communicative process should be taken into the consideration. PMID- 10689914 TI - [Phoniatric interpretation of evaluation principles in professional voice disorders]. AB - The critical assessment of the existing regulations of professional voice evaluation are presented. The necessity of the use of essential phoniatric principles is stressed in order to make respective legal decision more realistic and responsible. PMID- 10689913 TI - [Hearing loss, disorders of sound localization with the preservation ofsmell and taste in diabetics]. AB - Diabetes is often complicated by serious medical conditions which could be related to the development of auditory system and cranial nerves lesions, disorder of sound localisation and decreased olfactory and taste ability. Cranial nerve palsies in diabetes are considered as an integral part of the main disease. Twenty nine children with diabetes and non-diabetics control group, without a history of exposure to noise, ototoxic drugs, or ear disease aged 4-19 years old were examined. Hearing impairment may be present in children with disease duration above 3-5 years and in children with other complications. Sound localisation tests were performed by the method of Zakrzewski from a free auditory field, measuring the angle of directional hearing acuity in 35 diabetics age from 16 to 78 years. Longer duration of diabetes was associated with higher directional hearing acuity. Higher hearing loss was observed with higher directional values in diabetics. The authors investigated smell and taste in 35 diabetics. All patients were treated with insulin. Impairment value of smell identification thresholds was much more frequent than impairment of smell perception. In no case raised threshold of taste perception. PMID- 10689915 TI - [Phoniatric remarks on microsurgery of vocal cords]. AB - Own experience in microsurgery of vocal folds are presented. The necessity and importance of photographic and acoustic documentation (MDVP) as well as phoniatric pre- and postoperative rehabilitation are stressed. PMID- 10689916 TI - [The electrophysiological examination of hearing with some metabolic disorders in children]. AB - Electrophysiological examinations of hearing (ABR, SP) in 60 children with insulin-dependent diabetes mellitus and 39 children with obesitas simplex were done. In both groups with normal hearing threshold in pure tone audiometry disturbances in intrastem conduction were noticed. PMID- 10689917 TI - [Electrophysiological hearing examination in children and teenagers with insulin dependent diabetes mellitus]. AB - In 60 children and youngsters aged 14-19 (average 16.5) with insulin dependent diabetes (lasting on average 5.5 years) and normal hearing in pure tone audiometry electrophysiological examinations (ABR, SP) were performed; 48% of the cases revealed changes in latencies of I, III, V waves, especially as an elongation of I and V waves. In the peripheral interlatencie (III-I; V-I) shortening and in V-III elongation were observed. In SP examination most often shortening of N1 (67%) and P2 (87%) were found. Statistically significant dependence of latency changes on the state of metabolic balance was stated. It was not found referring to the duration of diabetes, age and gender of the patients. PMID- 10689918 TI - [The use of fibrin glue in otorhinolaryngology]. AB - Biochemical and physiological principles of biological sealing are presented. Various methods of application of biological seal and indications for its use in otorhinolaryngology are discussed. PMID- 10689919 TI - [The treatment of tracheal stenosis in Wegener's granulomatosis by laser surgery]. AB - A case of Wegener's disease is described. In eight years of remission a patient, treated with cyclophosphamid and steroids demonstrated sterosis of the trachea and upper way obstruction. We have performed carbon dioxide laser photoresection of granulotion mucosis and scars with an excellent results. PMID- 10689920 TI - [Tracheoesophageal fistula after the removal of esophageal foreign body]. AB - A case of 18 years old male with tracheoesophageal fistula as a result of esophageal foreign body (chestnut) is described. Foreign body was removed using esophagoscopy. Bronchofiberoscopy performed because of difficulties with swallowing and frequent airway infections revealed tracheoesophageal fistula. Fistula was closed from intratracheal access. Symptoms of dysfagia disappeared after surgery. PMID- 10689921 TI - [Pleomorphic adenoma of rare localization in the nose]. AB - Pleomorphic adenoma localised in nasal cavity is extremely rare. Authors described a case of mixed salivary gland tumor in the lateral wall of nasal cavity. We are presented, on the base of our experience and literature data, pathogenesis, clinical course and treatment of this tumors situated in atypical places. PMID- 10689922 TI - [Amyloidosis of the larynx]. AB - The paper presents three cases of localised amyloidosis of the larynx. The study was focused on case histories, findings of larynx examination and method of treatment was discussed. Amyloidosis is a disease characterised by gathering of amyloid deposits in extracellular space. The amyloid fibres consist of different kinds of proteins which have the ability of forming laminae. This disease may be secondary to numerous general processes in which amyloid proteins forms. Etiopathogenesis of this disease has not been explained yet. In the following paper the latest theories and classification of amyloidosis are presented. PMID- 10689923 TI - [Heterotopic salivary gland imitating laryngeal cyst]. AB - Patient aged 56 with heterotropic salivary gland in vestibule of the larynx was described. Localization of heterotropic salivary tissue in the larynx is very rare. PMID- 10689924 TI - [Thoracic duct cyst]. AB - Patient aged 48 with the cyst of the cervical part of thoracic duct was described. Cyst appeared as the result of inflammation. Histopathological examination excluded the cystic lymphangioma. PMID- 10689925 TI - [Sarcoidosis presenting as isolated nodules of the parotid and submandibular glands]. AB - The authors documented the case of healthy, 50 year old women with a sarcoid lesions of the parotid and submandibular gland without any other manifestations of the disease. The presence of noncaseating granulomas in salivary tissue is a specific feature of sarcoidosis. Primary involvement of the intraglandular lymph nodes with a secondary involvement of the salivary gland parenchyma was detected. Etiopathogenesis, diagnostics and clinical features of sarcoidosis were presented. PMID- 10689926 TI - [Craniofacial bone dysplasia]. AB - The case of fibrous dysplasia localised in the maxilla in 7 years old girl was described. Attention was paid to etiology, symptoms and treatment. The differential diagnosis was discussed. The diagnosis was wet set on the bases of clinical examination, X-ray, scyntygraphy and histological examination. PMID- 10689927 TI - [Mixed hearing loss in the case of Goldenhar-Gorlin syndrome]. AB - The authors describe a rare case of Goldenhar-Gorlin Syndrome, also called oculoauriculovertebral dysplasia (OAV) or hemifacial microsomia. Goldenhar's syndrome was accompanied by hearing loss caused by deformity of the auricle and atresia of the external auditory canal. PMID- 10689928 TI - [Nitric oxide: an important regulator of ciliary epithelial function of the upper respiratory tract]. AB - Importance of nitric oxide in regulation of ciliary activity and secretion of the mucin was discussed. Advantage of measurement of nasal nitric oxide concentration in course of maxillary sinuses diseases was emphasized. PMID- 10689929 TI - [The clinical features of cervical spondylosis with boule musculaire]. AB - To clarify the pathophysiology of boule musculaire associated with cervical spondylosis, we investigated the 13 patients (11 males and 2 females) with cervical radiculopathy caused by cervical spondylosis. None of the patients had any subjective symptoms such as sensory disturbance, weakness in the boule musculaire, and muscle atrophy of the insurround region near the boule musculaire. Among 21 tendon reflexes of the biceps muscle of the arm, 9 were normal, 8 were decreased and 4 were increased. Among the 13 patients there was only 2 clinical cases of cervical myelopathy. MRI obtained from 3 patients demonstrated a compressed cervical cord. Electromyography obtained from 7 patients demonstrated mild neurogenic changes such as polyphasic or prolonged motor unit potential (MUP) in the boule musculaire and in the atrophic regions. However, there was no giant MUP, fasciculation, fibrillation, nor positive sharp waves suggesting denervating lesions below the anterior horn in any patient. It was considered that the boule musculaire observed in cervical spondylosis gradually developed from a relatively mild disturbance of the ventral root. Moreover, the boule musculaire developed more often in the male patients, and predominantly on the right side. Cervical spondylosis accompanying boule musculaire may be a prodromal stage or a mild type of cervical spondylotic amyotrophy. PMID- 10689930 TI - [Coagulation and fibrinolytic activation in lacunar infarct patients]. AB - Lacunar infarcts are related to occlusion of penetrating arteries. Lipohyalinosis affects the smaller arteries 40-200 microns in diameter, and atherosclerosis involves larger arteries 200-850 microns in diameter. We hypothesized that the processes of thrombus formation might be different among these two kinds of lacuner infarcts, including those caused by lipohyalinosis and atherosclerosis. We studied acute coagulation and fibrinolytic activation in lacunar infarct patients which were divided into two groups according to their size: smaller lacunar group and larger lacunar group. Then we divided lacunar infarct patients into two groups in terms of the progression of motor deficits: those who showed the progression and those did not. And coagulation and fibrinolytic activation were compared each other. One hundred and twenty four patients were enrolled in this study, including 34 control subjects, 39 patients with smaller lacune (3 mm 10 mm in diameter), 28 patients with large lacune (10 mm-20 mm), and 23 patients with atherothrombotic infarcts confirmed by angiography. The levels of TAT activity in large lacune and atherothrombotic infarcts were significantly higher than those in control subjects (p = 0.009, p < 0.0001, respectively), whereas those in small lacune were not. Also, the levels of D-dimer activity in large lacune and atherothrombotic infarcts were significantly higher than those in control subjects (p = 0.0003, p < 0.0001, respectively), whereas those in small lacune were not. The progression of motor deficits were more frequently recognized in large lacune than in small lacune: three patients out of 39 small lacune patients and 22 patients out of 28 large lacune patients (difference was significant, p = 0.001). The level of TAT activity in patients who showed progression of motor deficits was significantly higher than that in those who did not (p = 0.0002), whereas the difference of the levels of D-dimer activity in two groups did not reach significant differencial levels. The process of thrombin and fibrin formation in large lacunar infarcts which are related to microatheroma and atheroscrelosis appears to be different from that in small lacunar infarcts. Antiplatelet and anticoagulation therapy should be tailored to large lacunar infarct patients. PMID- 10689931 TI - [Effect of prednisolone on apoptosis and cellular infiltration in mdx mouse muscle]. AB - Myonuclear apoptosis might precede the muscular degeneration of Duchenne muscular dystrophy. We examined influence of prednisolone (PSL) on apoptosis in the skeletal muscle of mdx mouse. Amount of apoptotic nuclei of the tibial anterior or quadriceps muscle was assessed by Tdt-mediated dUTP biotin nick end-labeling (TUNEL) method. When nuclei were classed into myonuclei and interstitial ones, apoptotic nuclei were observed mainly in the latter. In comparison between 7 and 33 week-old mice, interstitial apoptosis increased with age in both control and PSL group. Treatment with PSL decreased number of interstitial apoptotic nuclei. To the contrary, apoptotic myonuclei were increased with PSL treatment. Treatment with PSL significantly reduced focal accumulation of infiltrating mononuclear cells, possibly contributing to the suppression of muscle degeneration. Relationship between decrease of interstitial apoptosis and reduced cellular infiltration was unclear yet. Present study showed that PSL differently affected apoptosis between muscle cells and their interstitium. PMID- 10689932 TI - [Diagnosis of inflammatory myopathy; usefulness of 99mTc MDP scintigraphy and muscle MRI for determination of affected sites]. AB - We studied the effectiveness of 99mTc-MDP (methylendiphosphate) scintigraphy in imaging inflammatory myopathy. The three subjects including 1 male and 2 female patients had high creatine kinase (CK) levels and proximal dominant muscle weakness. In whole body muscle surveillance by 99mTc-MDP scintigraphy, abnormal 99mTc-MDP accumulation was found in the extremities of all patients. The sites with high 99mTc-MDP accumulation showed high intensity on T2 weighted MR imaging, suggesting an inflammatory process. Muscle biopsy was performed on two patients from the muscles with the abnormal MRI findings, which showed the diagnostic finding of inflammatory changes. Because muscle involvement in inflammatory myopathy differs from muscle to muscle, it is sometimes difficult to choose appropriate muscle biopsy sites for diagnostic purposes. Affected muscles are more easily identified by using 99mTc-MDP muscle scintigraphy and muscle MRI, therefore, a correct diagnosis and choice of biopsy site can be made. 99mTc-PYP scintigraphy is permitted for use in myocardial infarction alone and 111In antimyosin scintigraphy is not available in Japan. Therefore, we recommend 99mTc MDP scintigraphy for diagnosis of inflammatory myopathy and for determination of muscle biopsy sites. PMID- 10689933 TI - [An autopsy case of amyotrophic lateral sclerosis/parkinsonism-dementia complex with family history and living history in the Kii Peninsula focus]. AB - The patient was 68 year-old woman with an 8-year history of parkinsonism which was followed by psychiatric symptoms and neurogenic amyotrophy 5 years after the onset. She had a family history of parkinsonism associated with dementia in all of her three siblings. They grew up in Hobara village, a focus of amyotrophic lateral sclerosis (ALS) in the Kii Peninsula of Japan, in their childhood. Their parents were neither consanguineous nor natives of the Kii Peninsula. The brain weight was 1,040 g, and there were mild frontal lobe atrophy, moderate atrophy of pes hippocampi, decoloration of the substantia nigra and locus ceruleus, and anterior spinal root atrophy. The microscopic examinations revealed degeneration of CA 1 portion of the hippocampus to parahippocampal gyrus, substantia nigra, locus ceruleus and spinal anterior horn with Bunina body. The spinal pyramidal tracts also degenerated mildly. The neurofibrillary tangles (NFT) were observed in the cerebral cortex, especially in the cortices through hippocampus to lateral occipitotemporal gyri, basal nucleus of Meynert, basal ganglia, thalamus, substantia nigra and widespread regions of the central nervous system through the brainstem to the spinal cord including the nucleus of Onufrowicz. In spite of a few amount of the senile plaques in the cerebral cortex and Lewy bodies in the substantia nigra and locus ceruleus, abundant NFT distributed mainly in the third layer of the cerebral cortex, which is the characteristic finding of amyotrophic lateral sclerosis/Parkinsonism-dementia Complex (ALS/PDC) in the island of Guam. Thus this was verified ALS/PDC case outside the Guam island. The high incidence of neurological disease in her siblings and their history of living in the focus region in childhood suggest the genetic factor of ALS/PDC which is sensitive to certain environmental agents in the early stage of the life. PMID- 10689934 TI - [The cerebral peduncle lesion in multiple system atrophy]. AB - Atrophy of the cerebral peduncle and the pons can be seen on radiologic examination in patients of multiple system atrophy. This radiologic finding is one of the landmarks for the diagnosis of multiple system atrophy, but its detailed pathological background has not been thoroughly assessed. To clarify the pathological features of the cerebral peduncle lesion, samples from 28 autopsied cases (male 16, female 12; age 50-76 yr) were semiquantitatively examined after staining by HE, KB, Holzer, GFAP, Bodian and Gallyas methods. Atrophy of the cerebral peduncle was symmetric in most cases and resulted from the loss of small sized nerve fibers. The glial cytoplasmic inclusion (GCI) in the cerebral peduncle increased significantly in severe atrophic cases compared with mild atrophic cases. However, in the most severe atrophic cases, in which this tissue was severely damaged, the GCI was decreased. The atrophy of the cerebral peduncle correlated significantly with the degree of degeneration in the olivo-ponto cerebellar system and tended to correlate with a decrease in brain weight. The duration of disease is significantly longer in severe atrophic cases compared with mild atrophic cases. PMID- 10689935 TI - [A case with HTLV-I associated myelopathy (HAM) accompanied by primary biliary cirrhosis (PBC) and autoimmune hepatitis (AIH)]. AB - We reported a 60-year-old female patient with HTLV-I associated myelopathy (HAM) accompanied by primary biliary cirrhosis (PBC) and autoimmune hepatitis (AIH). The diagnosis of PBC and AIH was confirmed by liver biopsy. HAM is considered to be mediated by cellular immune mechanisms, while humoral immune mechanisms may play a predominant role in the development of PBC and AIH. Flowcytometric analysis of lymphocyte subset of peripheral blood was within normal limits. We then collected CD4 positive cells from the patient. These cells expressed T helper 2 (Th 2) cytokine mRNA such as IL-4 and IL-10, but did not express Th 1 cytokines, indicating the predominance of Th 2 in this patient. This case suggested the possibility that disease associated Th 2 might develop in the course of Th1-mediated disease like HAM. PMID- 10689936 TI - [A family with probable autosomal dominant bulbospinal muscular atrophy with gynecomastia]. AB - We reported a 52-year-old man and his family with bulbospinal muscle atrophy (BSMA) and gynecomastia. The propositus presented with the clinical picture of late onset progressive bulbospinal muscular atrophy including postural tremor, general hyporeflexia, mild maturity onset diabetes, gynecomastia and sexual impotence. One of his brother and his two sons had gynecomastia. His elder son had ocular movement abnormality, associated movement of facial muscle and finger tremor. One of his brothers showed tongue fasciculation without gynecomastia. None of members examined had abnormal expansion of CAG repeats in the androgen receptor gene. We speculate that this family has a new clinical entity characterized by bulbospinal muscular atrophy with an autosomal dominant inheritance. PMID- 10689937 TI - [A novel splice-site mutation in the STA gene in a Japanese patient with Emery Dreifuss muscular dystrophy]. AB - Emery-Dreifuss muscular dystrophy (EDMD) is an X-linked recessive or autosomal dominant progressive muscular dystrophy characterized by progressive muscle wasting and weakness with scapulo-humero-peroneal distribution, early contracture and cardiomyopathy with conduction block. The responsible gene for EDMD, designated as 'STA', has been mapped to Xq 28 and cloned. It encodes a serine rich protein of 254-amino-acid, called 'emerin', localized in the inner nuclear rim. We performed genetic analysis of a 23-year-old male clinically diagnosed as EDMD and found a novel point mutation. Total RNA was extracted from skeletal muscle and reverse-transcription and polymerase chain reaction amplification was performed using a set of oligonucleotide primers between 5'-flanking site of exon 1 and exon 4. Our patient gave a smaller PCR product (about 30 bp) than normal control. The determined cDNA sequence revealed a deletion of 29 bp, spanning position 164 to 192 in exon 1. To clarify the mutant allele, we performed genomic DNA sequence. Genomic DNA sequence from the initiation of exon 1 to the upstream lesion of exon 2 confirmed a novel point mutation G to C, at nucleotide 197 in the donor splice site of intron 1. This point mutation may interfere with the correct splicing of the mRNA and cause frameshift, resulted in truncation of predicted protein by premature stop. We report a novel point mutation G to C, at nucleotide 197 in the intron 1 of STA gene corresponding the truncation of predicted protein, which differs from any of the previously reported mutations. PMID- 10689938 TI - [A case of adult cerebral X-linked adrenoleukodystrophy (X-ALD) accompanying typical hypertrophic neuropathy with marked onion-bulb formation]. AB - We report a case of adult cerebral X-linked adrenoleukodystrophy (X-ALD). His illness started with minimally unsteady gait and slight difficulty in speaking at age 43; then mental deterioration progressed rapidly within two years, which was the characteristic clinical feature of this case. He had not shown any signs and symptoms of spastic paraparesis and adrenal dysfunction. Sural nerve biopsy revealed marked demyelination and moderate loss of nerve fibers, showing typical hypertrophic neuropathy with marked onion-bulb formation, while trilaminar inclusions considered as pathognomonic to X-ALD were not detected in our case. Although onion-bulb formation has sometimes been referred to in the peripheral nerve specimens from X-ALD patients, such a remarkable onion-bulb formation as seen in this case is extremely rare. It suggests that severe destruction of myelin in the peripheral nerve have persisted for a fairly long period without drawing any attention from medical staff. PMID- 10689939 TI - [A case of neuroleptic malignant syndrome associated with reversible leukoencephalopathy]. AB - We report a case of neuroleptic malignant syndrome associated with reversible leukoencephalopathy. The patient was a 60-year-old woman. Soon after ingesting an antipsychotic drug, the patient developed neuroleptic symptoms. After hydration and dantrolene sodium were administered, muscular rigidity gradually improved and serum levels of CK became normal. On the 7th hospital day, however, she fell into coma and showed tetraplegia. Although brain CT was normal on admission, diffuse low density areas were observed on the parieto-occipital cerebral white matter. The same lesions were observed on T2-weighted MRI. On the 20th hospital day, after giving her glycerol and adrenocorticosteroid, not only abnormal neurological findings but also abnormal CT and MRI findings disappeared. There were only two case reports of leukoencephalopathy with malignant syndrome in the literature. This is a rare case of reversible leukoencephalopathy with neuroleptic malignant syndrome due to the antipsychotic drug. PMID- 10689940 TI - [A case of cerebellar degeneration showing amaurosis fugax due to primary angle closure glaucoma]. AB - We reported a 76-year-old woman with cerebellar degeneration who had transient monocular visual loss following the acute attacks of angle-closure glaucoma. The episodes occurred only at night approximately every ten days. She denied pain or any other associated symptoms. Ophthalmological examinations including intraorbital pressure, ocular fundus, visual acuity and visual field showed no abnormalities between the attacks. Provisional diagnosis on admission was amaurosis fugax from retinal embolization. After admission, she developed a typical acute attack of glaucoma accompanied by severe pain in her left eye. Intraorbital pressures were 12 mmHg in the right eye and 58 mmHg in the left, and the diagnosis of primary angle-closure glaucoma was made gonioscopically. Following peripheral iridotomy by laser therapy, her visual acuity recovered and episodes of visual loss disappeared. In this case, the attacks of glaucoma were unusually painless, so it is very difficult to distinguish between glaucoma and amqurosis fugax from retinal embolization. The transient visual loss always occurred at night, and retrospectively, this characteristic feature might indicate that these episodes were acute attacks of angle-closure glaucoma. Glaucoma is one of the diseases that can cause painless amaurosis fugax. PMID- 10689941 TI - [A Japanese case of Poland-Mobius syndrome]. AB - We herein report a Japanese case of Poland-Mobius syndrome. The patient was a 19 year-old female. She was the product of a full-term forceps delivery. Birth weight was 2500 g. She had a defect of the right pectoral muscle, and syndactyly of the right hand. When she was 10 days old, facial diplegia, bilateral abducens nerve palsy, and bilateral ptosis were also noted. She was admitted to our hospital at 19 years of age. On physical examination, she had microsyndactyly of the right hand, and her right pectoralis major muscle was absent. Neurological examination revealed bilateral abducens nerve paresis, mild impairement of the upward and adducting movement of both eyes and bilateral facial weakness and atrophy of the left side of her tongue. Her karyotype was normal. Neither R 1 nor R 2 response was evoked in the blink reflex on either side. Brain MRI disclosed thin facial nerves and atrophy of the pons and medulla. Therefore, she was diagnosed as a case of Poland-Mobius syndrome. In this case, the facial nerves were considered to be hypoplastic. PMID- 10689942 TI - [A case of Sweet's syndrome (acute febrile neutrophilic dermatosis) showing transient jargon aphasia]. AB - A 54-year-old woman visited our emergency service complaining of a severe language disturbance. She was fluently speaking something but most of the words were merely meaningless syllables. This jargon state lasted only four hours, then her abnormal speech rapidly and completely recovered within 24 hours. She had also suffered from painful oral ulcers, fever and erythema-like eruptions on her face for about three weeks. Skin biopsy of a facial lesion revealed a dense infiltrate of neutrophils in the dermis and minimal features of vasculitis, which, with other typical clinical findings, led us to the diagnosis of Sweet's syndrome. Although head CT scans, MRIs, MRA or SPECT could not detect any brain lesions, a cerebrospinal fluid examination showed a mild pleocytosis of 38/mm3 with 47% polymorphonuclear cells. There have been a few case reports on Sweet's syndrome accompanying neurologic symptoms, most of which are mild meningitis. We speculate that the transient aphasia was due to a focal lesion in the central nervous system attributable to Sweet's syndrome. Sweet's syndrome might bring inflammatory or ischemic lesions to the cerebrum as Behcet's disease. PMID- 10689943 TI - [A case of non-Hodgkin lymphoma in the central nervous system, developing during treatment of galactorrhea amenorrhea syndrome]. AB - We report a 27-year-old woman with a non-Hodgkin lymphoma in the central nervous system (CNS), showing monoparesis of the right upper extremity during treatment for the galactorrhea amenorrhea syndrome. The MRI demonstrated an infiltrative lesion with an obvious gadolinium-enhancement effect, extending from the pituitary stalk and hypothalamus through the 4th ventricle to the dorsal part of the medulla and upper cervical spinal cord. Because no tumors were detected in any other organ in either a physical examination or radiological investigations, including CT for the chest and abdomen, she was diagnosed as having primary CNS non-Hodgkin lymphoma (B cell origin) on the basis of an open brain biopsy. After the irradiation of the whole brain, followed by chemotherapy (methotrexate + CHOP), the infiltrative tumor disappeared on the MRI with a slight improvement for monoparesis of the right upper extremity. In this patient, primary CNS lymphoma might originate around the hypothalamus and infiltrate the medulla, inducing not only monoparesis of the right upper extremity but the galactorrhea amenorrhea syndrome. PMID- 10689944 TI - [Direct Gram staining of blood culture sample enabled the early diagnosis of brain abscess due to Listeria monocytogenes]. AB - A 58 year old woman had a long history of immunocompromised state. Since age 28 she had multiple endocrine neoplasm type 2A: her thyroid gland and bilateral adrenal glands were removed because of pheochromocytoma and thyroid medullary carcinoma. Corticosteroid and levothyroxine were supplemented. At age 57 she was afflicted with systemic lupus erythematodes and nephrotic syndrome. Prednisolone therapy was started. Two months later she developed fever, lethergy, headache and left hemiparesis. MRI revealed multiple ring-enhancing lesions in the right cerebrum. CSF was negative for microorganisms. Blood culture hemolysed after 24 hours. Direct gram staining of the blood culture sample revealed gram-positive short rods without spore, suggested listeria. This enabled prompt initiation of high dose penicillin therapy before the official report of listseria infection. Neurological abnormality including left hemiparesis disappeared completely within one month. Enhancement of abscess wall decreased every month, but it persisted for five months despite continuous intravenous penicillin therapy. Listeria monocytogenes is well-recognized as an opportunistic pathogen. It requires prolonged therapy with antibiotics, since it is the intracellular organism. Monitoring of the brain abscess wall by the enhanced MRI is useful to determine the completion of therapy. Since listerial contamination is common among raw meat and unpasteurized milk, immunocompromised patients should be alarmed not to eat uncooked food products. PMID- 10689945 TI - [Appropriate method of apheresis in Guillain-Barre syndrome]. PMID- 10689946 TI - [Dementia with Lewy bodies and Parkinson's disease with dementia]. PMID- 10689947 TI - [Helicobacter and malignant diseases of the ventricle]. PMID- 10689948 TI - [Influenza--drug therapy and prevention]. PMID- 10689949 TI - [Use of glucocorticoids in rheumatoid arthritis]. AB - OBJECTIVES: To determine whether short-term (i.e. as recorded within the first month of therapy), oral low-dose corticosteroids (corresponding to a maximum of 15 mg prednisolone daily) is superior to placebo and nonsteroidal, antiinflammatory drugs in patients with rheumatoid arthritis. SEARCH STRATEGY: Medline Silverplatter, The Cochrane Controlled Trials Register, reference lists and a personal archive. SELECTION CRITERIA: All randomised studies comparing an oral corticosteroid (not exceeding an equivalent of 15 mg prednisolone daily) with placebo or a nonsteroidal, antiinflammatory drug were eligible if they reported clinical outcomes within one month after start of therapy. DATA COLLECTION AND ANALYSIS: Decisions on which trials to include were made independently by two observers based on the methods sections of the trials only. Standardised effect measures were used for the statistical analyses; the random effects model was used if p < 0.10 for the test of heterogeneity. MAIN RESULTS: Ten studies, involving 320 patients, were included in the meta-analysis. Prednisolone had a marked effect over placebo on joint tenderness (standardised effect size 1.31, 95% confidence interval 0.78 to 1.83), pain (standardised effect size 1.75, 0.87 to 2.64) and grip strength (standardised effect size 0.41, 0.13 to 0.69). Measured in the original units, the differences were 12 tender joints (6 to 18) and 22 mmHg (5 to 40) for grip strength. Prednisolone also had a greater effect than nonsteroidal, antiinflammatory drugs on joint tenderness (standardised effect size 0.63, 0.11 to 1.16) and pain (standardised effect size 1.25, 0.26 to 2.24), whereas the difference in grip strength was not significant (standardised effect size 0.31, -0.02 to 0.64). Measured in the original units, the differences were 9 tender joints (5 to 12) and 12 mmHg (-6 to 31). The risk of adverse effects, also during moderate- and long-term use, seemed acceptable. CONCLUSIONS: Prednisolone in low doses (not exceeding 15 mg daily) may be used intermittently in patients with rheumatoid arthritis, particularly if the disease cannot be controlled by other means. Since prednisolone is highly effective, short-term placebo controlled trials studying the clinical effect of low-dose prednisolone or other oral corticosteroids are no longer necessary. PMID- 10689950 TI - [Invasive aspergillosis in hematological patients]. AB - An increasing incidence of invasive aspergillosis over the last decades is well documented in patients with haematological malignancies and is the most significant fungal infection in patients undergoing bone marrow transplantation and in aplastic anaemia. The diagnosis is difficult as clinical signs and symptoms usually are non-specific, but can be supported by frequent radiological examinations of the chest and sinuses and successive demonstrations of Aspergillus antigen in serum. The prognosis depends on the course of the underlying disease. A regeneration of the neutrophil granulocyte number is a condition for successful treatment. Early antifungal therapy is often necessary in neutropenic patients with fever and a lung infiltrate that does not remit following broad spectrum antibacterial treatment. Because of the risk of relapse after successful treatment these patients should receive prophylactic antifungal treatment during subsequent neutropenic episodes. PMID- 10689951 TI - [Prognosis after late versus early nonfatal myocardial infarction]. AB - Many recent studies have identified nonfatal recurrent myocardial infarction (RNMI) as the most significant predictor for later outcome. Almost all of these studies have been based on the studies of RNMI in the first year after the index infarction. The prognosis after late RNMI has not been studied properly. In 3,867 nonselected patients below 76 years of age with an acute myocardial infarction we studied the prognosis after a first RNMI depending on the year of its occurrence after the index infarction. Mortality was estimated by the method of Kaplan-Meier and the differences were evaluated by means of the Tarone-Ware test. Four hundred and ninety-three (13.6%) patients had a first RNMI in the first, 151 (5.4%) in the second, 105 (4.2%) in the third, and 71 (3.8%) in the fourth year after the index infarction (group 1-4). One-year mortality rate after RNMI was 23.7% in the first group, 24.1% in the second, 17.5% in the third, and 22.8 in the fourth group. When all the groups were compared with each other no significant difference was found between the mortality rates (p = 0.12) or Standardised Mortality Rates. We concluded that late and early RNMIs have almost the same grave prognosis. PMID- 10689952 TI - [Detection of cholangiocarcinoma in primary sclerosing cholangitis by positron emission tomography]. AB - Primary sclerosing cholangitis (PSC) predisposes to cholangiocarcinoma (CC). PET scanning can assess metabolism in vivo. The glucose analogue [18 F]fluoro-2-deoxy D-glucose (FDG) accumulates in malignant tumours because of high glucose metabolism. PET scanning of the liver was performed after intravenous FDG in nine patients with PSC, six with PSC + CC, and five controls. "Hot spots" with radioactivity accumulation were seen in each PSC + CC patient, but not in the two other groups. Values of net metabolic clearance of FDG, K (ml min-1 100 ml-1 tissue), was in CC hot spots 1.59 to 4.17 (median, 2.34; n = 6); in reference liver tissues of these patients 0.40 to 0.69 (0.49); in PSC 0.23 to 0.53 (0.36); in controls 0.20 to 0.34 (0.31). The difference between K in CC hot spots and the other groups was statistically significant (P < 0.001). FDG-PET may detect small CC tumours and be useful in therapeutic management of PSC. PMID- 10689953 TI - [The staff's knowledge of patients' social function and needs--in connection with discharge planning]. AB - The aim of the study was to assess the staff's knowledge about the social functioning and needs of the patients. A cross-sectional study interviewing 48 staff members using the Camberwell Assessment of Need--CAN was carried out. In nine of CAN's 26 areas more than 5% of the staff did not know if the patient had a problem. Among the patients with a problem more than 5% of the staff could not assess the need for care in four areas. The staff generally had a good knowledge concerning the patients' need for care. CAN seem to be a relevant instrument for the purpose of discharge planning. PMID- 10689954 TI - [Pseudolymphoma and ventricular maltoma in patients with chronic gastritis, ulcer and Helicobacter pylori infection]. AB - Among 128 patients with malignant B-lymphoproliferative disorders, 19 patients had long lasting dyspepsia and gastroscopy showed chronic active gastritis or gastric ulcer. PCR analysis for TCR and IgH clonality in biopsies showed local involvement of the malignant lymphocyte clone in four patients out of eight indicating presence of these cells in the inflammatory infiltrate. Weak B-cell clonality was found in four patients. A close relationship was seen between lymphocytic clonality and immune response to H. pylori Cag A, and all patients had parietal cell antibodies. Thus, the malignant clone may participate in the local inflammatory reaction, and continued local stimulation by H. pylori as well as parietal cell antigens may lead both to autoimmunity as well as a clonal development of lymphocytes. PMID- 10689955 TI - [Invasive aspergillosis in two hematological patients]. AB - We present two cases of invasive aspergillosis in patients with acute myeloid leukemia. In neutropenic patients with antibiotic resistent fever, without specific symptoms or signs, invasive aspergillosis should be considered. Diagnostic approaches such as X-ray/CT-scan of the thorax and sinuses, relevant cultures and antigen detection should be performed. Due to diagnostic difficulties and the rapid progression of the infection empirical antifungal therapy should be given. During subsequent neutropenic episodes prophylactic antifungal therapy can possibly preempt recurrence. PMID- 10689956 TI - [Massive gastrointestinal hemorrhage from the jejunum caused by Dieulafoy's lesion]. AB - A 54-year old woman was admitted to the hospital because of massive haematochezia. Emergency surgical exploration was performed and by a simple method the source of bleeding was localized and treated. We discuss one of the major problems in all GI-bleeding that lies in establishing the diagnosis. With localization in the small bowel the problem is even bigger because this part of the bowel isn't examined by conventional endoscopy. This case-story should be a reminder of the small bowel as a source of bleeding when conventional upper and lower endoscopy does not reveal the bleeding source. PMID- 10689957 TI - [Dampness in an electric plug as a cause of electricity failure in an operation theatre]. AB - Two cases of electricity failure in an operation theatre during open heart surgery are discussed. The fuse for the patient monitor, ventilator, surgery instruments and heart lung machine was blown. Short-circuit was established because of humidity in the plug of the heater for fluid and blood. We recommend sealed or founded plugs and that anaesthesia equipment should not be used as an electrical supply for other electronic apparatus. PMID- 10689958 TI - [Ethnicity and the course of psychosis]. PMID- 10689959 TI - [The difficult medical profession]. PMID- 10689960 TI - [Pagers and breast cancer among female physicians]. PMID- 10689961 TI - [Total synthesis of crassostreaxanthin B directed toward the biomimetic synthesis of carotenoids]. AB - Marine carotenoids halocynthiaxanthin 2, mytiloxanthin 3 and crassostreaxanthin B 4 have characteristic structures, commonly possessing a monoacetylenic end group. The cyclopentyl end group of 3 is believed to be formed in nature from the epoxide end group of 5,6-epoxy carotenoids such as 2 (Chart 1, route a). It is also conceivable that 4 including a novel tetrasubstituted olefinic end group arises from epoxy carotenoids by the opening of the C-6-oxygen bond of the oxirane ring and the subsequent migration of the methyl group at the C-1 position (route b). We found that treatment of the epoxide 6a having a partial structure of epoxy carotenoids with Lewis acids gave the cyclopentyl ethyl ketone 9 possessing the same configuration as 3, and the acyclic tetrasubstituted olefinic methyl ketone 11 including a partial structure of 4 (Chart 2). It supported the proposed metabolic pathway of 5,6-epoxy carotenoids. Toward the biomimetic synthesis of 4, we examined the reaction of epoxides having several substituents at C-6 position with Lewis acids. Among these epoxides, an epoxide 60 was found to provide a tetrasubstituted compound 61 as a major product. This could be converted into an aldehyde 73 in 8 steps which was transformed into a compound 75 through the coupling reaction with vinyllithium 63. Then, the first total synthesis of 4 was accomplished via the double Wittig condensation of 75 with phosphonium salts 76 and 77. PMID- 10689962 TI - [Syntheses and properties of conformationally restrained nucleosides and oligonucleotides analogues]. AB - This review summarizes our efficient syntheses of novel bicyclic nucleoside analogues, 3'-O,4'-C-methyleneribonucleosides (1) (4-BC type nucleoside analogue), 2'-O,4'-C-methyleneribonucleosides (2) (5-BC), 3'-amino-3'-deoxy-3' N,4'-C-methyleneribonucleosides (3) (aza 4-BC), and 3'-azido- and 3'-amino-3' deoxy-2'-O,4'-C-methyleneribonucleosides (4, 5) (aza 5-BC). From 1H-NMR and X-ray crystallographic analyses, the 4-BC and aza 4-BC type nucleoside analogues (1, 3) were found to have a S-conformation predominantly, while the conformations of 5 BC and aza 5-BC type nucleoside analogues (2, 4, 5) were exclusively locked in N form. The 4-BC and 5-BC type nucleoside analogues (1, 2) were effectively introduced into oligonucleotides using a DNA synthesizer. Furthermore, unprecedented hybridizing ability towards complementary RNA and DNA, RNA selectivity, potent triplex forming ability, and sufficient enzymatic stability of these modified oligonucleotides were also confirmed. These results should reveal a promising route to the development of antisense/antigene methodology. PMID- 10689963 TI - [Total analysis system for tumor promoter microcystins produced by cyanobacteria]. AB - Microcystins, produced by freshwater cyanobacteria, are cyclic peptide hepatotoxins and tumor promoters. An outbreak of human poisoning attributed to microcystins has been reported in Caruaru, Brazil in 1996, where exposure through renal dialysis led to the death of 50 patients. Although such severe acute effects on human health seem to be rare, microcystins poses problems to human health which could result from low-level, chronic exposure to microcystins in drinking water. It is therefore important to monitor the levels of microcystins in water reservoirs where cyanobacterial blooms occur. We have developed a total analysis system for microcystins using GC-MS and LC-MS. This comprises initial screening of samples to check for the presence of microcystins by detecting 2 methyl-3-methoxy-4-phenylbutyric acid as an ozonolysis product using thermospray interface LC-MS and electron ionization/GC-MS. If a sample is positive in a screening test, it will be necessary to follow through with identification and quantification. Frit-FAB interface LC-MS allowed the rapid identification of microcystins in cyanobacteria and lake water, and also enabled us to identify microcystins and their metabolites formed in vivo in mouse liver. Finally, Frit FAB/LC-MS using selected ion monitoring could be used for quantitative analysis of microcystins in lake water in the low nanogram range. The total analysis system proposed in the present study should be applicable to studies of the metabolism of microcystins, of their detoxification, and those of the mechanism(s) of the accumulation in the food chain. PMID- 10689964 TI - [Preparation of chemically modified carbon electrodes by anodization in 1 alkanols and their application to electrochemical analysis]. AB - Anodization of a glassy carbon (GC) in a 1-alkanol in a cycled or constant potential mode serves as a useful tool for preparing a chemically modified GC electrode. By this treatment, 1-alkanol molecules are fixed on the GC surface via ether linkage. As the 1-alkanol in the anodic modification, CH3(CH2)nOH (1: n = 0 7), HO(CH2)nOH (2: n = 1-5), and HO(CH2CH2O)nR (3: n = 1-4, R = H; 4: n = 1-3, R = CH3) are utilized. The surface of a GC electrode anodized in the 1-alkanol remarkably reflects the identities of the modifiers. Some of the modified GC electrodes exhibit surface characteristics useful for electroanalytical application as follows: (1) the surface of the GC electrode anodized in 3 or 4 is hydrophilic and resists protein adsorption. An HPLC system equipped with an electrochemical detector employing the GC plate anodized in triethylene glycol as a working electrode has proven to provide a useful analytical method for a protein-containing sample; (2) in the course of anodization of the GC electrode in 2, the diol molecules are first fixed on the surface via ether linkage with one of hydroxyl groups, and the remaining terminal hydroxyl groups in some of the fixed molecules are then oxidized to carboxyl groups. Thus, the GC electrode anodically modified with 2 has carboxyl groups on the surface, which allow dopamine to be voltammetrically discriminated from ascorbic acid in a large excess; (3) when the GC electrode is anodized in triethylene glycol containing HOCH2CH2SO3Na, carboxyl groups are effectively introduced on the surface. On the basis of the formation of an amide bond formation through chemical reaction between the functional groups and amino compounds, electrochemical catalysts such as 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO) and catechol are immobilized on the surface of the GC electrode. The obtained electrodes shows stable voltammetric and electrochemically catalytic performance probably because the catalytic molecules are confined on the electrode surfaces via a hydrophilic linker instead of a hydrophobic one. PMID- 10689965 TI - [Molecular design and control of functional expression of ate complexes]. AB - As a new type of zincate, "highly coordinated" zincates, Me3Zn(R)Li2 (R = Me, CN, SCN) were designed. On the basis of their excellent chemical yields and chemoselectivities, these species were considered to be differentiated from ordinary triorganozincates, R3ZnLi. The structures of the newly designed zincates were discussed on their spectroscopic studies. All results strongly support the fact that these newly designed zincates are new category of zincate species. Various dialkylzinc hydride "ate" complexes were also designed and the reactivities of these zincates toward the carbonyl compounds investigated. The results clearly reveal that dimethylzinc hydrides are the most powerful and selective zincate for the reduction of the carbonyl group. PMID- 10689967 TI - [Studies on the syntheses and reaction of nitrogen-containing heterocyclic compounds centered the naphthyridines]. AB - 1, X-Naphthyridines (X = 5, 6, and 8) (1-3) were synthesized in a high yield by the Skraup reactions of 4-, 3-, and 2-aminopyridines with glycerol, in the presence of ferrous sulfate and boric acid. The improved syntheses were applied to the syntheses of 1,5-, 1,6-, and 1,8-naphthyridines (4-28), pyridonaphthyridines (31, 32), benzonaphthyridines (38-42), naphthonaphthyridines (44, 48, 49, 59). New synthetic methods of naphtho[1,2-b or 2,1 b][1,8]naphthyridine (62, 63), benzo[b][1,8]naphthyridine (41) and benzo[g]quinoline (67) were developed and their compounds obtained conveniently. Reissert reaction of 1,6-naphthyridine (2) using triethylbenzylammonium chloride (TEBA) produced Reissert compounds and ring-opened compounds. Reissert reaction of 1,7-naphthyridine (29), 4,7- (87), 4,6-phenanthroline (38) gave the Reissert compounds, and the compounds (2, 38) gave their pseudo bases (79, 90). Reissert type reactions of benzo[f]quinoline (68), and 1,7-phenanthroline (105) with acyl chloride and phosphite, gave alpha- and gamma-phosphonate, and 87 produced alpha,alpha'-(108a-c) and alpha,gamma'-diphosphonates (109a-c). The structure of 108c-1 was determined to be trans-type by an X-ray analysis. Compounds 1-3, 29, and benzo[f or h]quinoline were treated with dimetyl sulfoxide in the presence of sodium hydride to give mainly methylated compounds at the position para to the ring-nitrogen. When benzo[f or h]quinoline N-oxide was treated with methylsulfinylmethyl carbanion in the usual procedure, a new reaction took place to produce phenanthrene in an excellent yield. The same reaction of 1,10-, 1,7-, or 4,7-phenanthroline N-oxides (143-145) resulted in the liberation of the N oxide group to form benzo[f or h]quinoline, but isoquinoline N-oxide afforded to benzazepine derivatives (161). Reaction of quinaldine with methylsulfinylmethyl carbanion gave novel tricyclic compound (121a). The oxidation of 41, 62, and 63 with peroxy acids afforded novel products such as seven-membered 1,4-oxazepine derivatives. PMID- 10689966 TI - [A novel synthetic approach of tryptophan-containing cystine peptides by regioselective disulfide bond-forming reaction using the silyl chloride-sulfoxide system]. AB - A general scheme for the efficient synthesis of Trp-containing cystine peptide by the successive treatment with methyltrichlorosilane-diphenylsulfoxide in trifluoroacetic acid (TFA) and trifluoromethanesulfonic acid (TFMSA)-thioanisole in TFA, is described. A disulfide bond-forming reaction by silyl chloride sulfoxide system is completed within 10-15 min without modifications at sensitive residues (Tyr, His, Met) in peptide chain, except for a Trp residue. In order to synthesize a Trp-containing cystine peptide using silyl chloride, the indole moiety of Trp has to be protected since the chlorination of the indole ring proceeded predominantly. A formyl group has been the only protecting group employed for this purpose in practical syntheses of cystine peptides, although it was clarified that a side reaction derived from the formyl group migration was inevitable in the synthesis of somatostatin. Firstly, we examined the application of the following Nin-protecting groups, mesitylene-2-sulfonyl (Mts), cyclohexyloxycarbonyl (Hoc), and 2,4-dimethylpent-3-yloxycarbonyl(Doc) for an efficient synthesis of the Trp-containing cystide peptide by the silyl chloride method. In order to find a feasible scheme of the successive treatment with CH3SiCl3-PhS(O)Ph/TFA and TFMSA-thianisole in TFA, we synthesized somatostatin using Trp(Mts), Trp(Hoc) or Trp(Doc) derivative. The Doc group was found to be the most suitable as an indole protecting group, since the protecting group was cleaved under mild conditions (4 degrees C, 30 min) via the corresponding Nin carboxylic acid intermediate. We then applied the above procedure to the synthesis of endothelin-1 (ET-1), a peptide containing 21-amino acid residues having a C-terminal Trp residue and two disulfide bonds, by regioselective disulfide formation. The combination of the silyl chloride method with iodine oxidation using S-acetamidomethyl (Acm) and S-tBu groups for the regioselective double disulfide formation was successfully applied to give a highly purified ET 1. These results also show that the Nin-Doc group would be useful for the efficient syntheses of complex cystine-peptides by the silyl chloride method. PMID- 10689968 TI - [Drug Compliance Scale. I. Development of the Drug Compliance Scale]. AB - The failure of patients to comply with treatment regimens recommended by their physicians is a significant clinical problem. Researches on the assessment of compliance have, however, been precluded by methodological difficulties such as lack of adequate measures. The purpose of this study was to develop a self administered questionnaire to evaluate drug compliance. First, questionnaire containing a 52-items complied by two doctors, a pharmacist and a nurse, was tested on 81 outpatients, all volunteers, attending the departments of psychosomatic medicine and internal medicine. Four items were temporarily removed for later analysis because they directly inquired about drug compliance (drug compliance items). The other 48 items were analyzed and three factors consisting of 26 items were further studied: expectation on taking medicine, rejection to taking medicine and seeking knowledge of drugs. Chronback's alpha coefficients representing internal consistency of the three factors were sufficiently high (ranging from .75 to .84). Furthermore, we preformed a simplified pill count to validate the 4 drug compliance items. There was a weak to moderate correlation between the result of pill count and each of 4 drug compliance items. A new self administered questionnaire of 30 items was thus developed and named the Drug Compliance Scale. PMID- 10689969 TI - [Drug Compliance Scale. II. Psychological factors affecting drug compliance in the department of psychosomatic medicine]. AB - The purpose of this study was to investigate psychological factors affecting drug compliance in the department of psychosomatic medicine. Seventy-four outpatients were asked to answer a battery of self-administered questionnaire including the Drug Compliance Scale (DCS) that we had recently developed and other questionnaire evaluating psychological and vegetative symptoms, self-efficacy and attributional styles on the promotion of health and personality closely related to interpersonal relationships. Results of path analysis indicated that attributional styles and self-efficacy mainly affected three factors of DCS such as expectation on taking medicine, rejection to taking medicine and seeking knowledge of drugs, through which they influenced drug compliance, and also indicated that personality and self-efficacy mainly affected the stability of mood state, suggesting a further influence on drug compliance. PMID- 10689970 TI - [Investigation on the marker substances of crude drugs in formulations. II. Marker substances for the identification of Cistanchis Herba in drinkable preparations]. AB - There are few reports about marker substances for the identification of Cistanchis Herba in formulations. First, constituent analysis was performed by HPLC for screening of a marker substance, using several lots of Cistanchis Herba and its fluid extract. As a result, two components were clearly detected, which were thought to be good marker substances and identified to be echinacoside and acteoside by structural analysis. Stability testing of these two marker substances in various pH and temperature conditions was carried out, which suggested that they are stable and suitable enough for the identification. Therefore, the identification methods of Cistanchis Herba and its fluid extract in formulations were investigated using five different kinds of commercial drinkable preparations with authentic standard of echinacoside and acteoside as marker substances. Consequently, echinacoside and acteoside were clearly detected in all formulations investigated, using an HPLC-photodiode array detector system. Echinacoside and acteoside will be successfully used as marker substances for the identification of Cistanchis Herba and its fluid extract in formulations. PMID- 10689971 TI - Hundreds of RNs across the country showed that nurses CAN. PMID- 10689972 TI - Initiating a dialogue with the adolescent patient. PMID- 10689973 TI - New law eases Medicare interim payment system burden on home health care. PMID- 10689974 TI - Nurse-midwife empowers inner-city women through birth center [interivew by Michelle Slattery]. PMID- 10689975 TI - ANA media outreach makes a difference. PMID- 10689976 TI - TNA "gun safe" campaign unites community with common goal. PMID- 10689977 TI - The human connection: nurses and their patients. PMID- 10689978 TI - Protecting the patient by protecting the worker. PMID- 10689979 TI - Why don't more African Americans go into nursing? One experience. PMID- 10689980 TI - Providing primary care the NP way. PMID- 10689981 TI - Holistic nursing: the goal is the whole person. PMID- 10689982 TI - Workplace violence affects one-third of nurses. PMID- 10689983 TI - Workplace violence: one nurse's night of fear. PMID- 10689984 TI - RNs confront causes, consequences of school violence. PMID- 10689985 TI - Nevada nurses secure patient protections. PMID- 10689986 TI - Health care community faces year 2000. ANA joins group addressing patient care concerns. PMID- 10689987 TI - Perceived ability to manage juvenile chronic arthritis among adolescents and parents: development of a questionnaire to assess medical issues, exercise, pain, and social support. AB - OBJECTIVE: To develop a questionnaire to assess perceived ability to manage juvenile chronic arthritis (JCA) among adolescents and parents. METHODS: The questionnaire contained 24 (parents' version) and 23 (adolescents' version) questions accompanied by visual analog scales in the areas of knowledge, skill, behavior, attitudes, and self-efficacy. One hundred seven persons participated in the examination of validity and 25 in the examination of test-retest reliability. RESULTS: Factor analyses indicated that the questionnaire, now termed the MEPS (abbreviation for "medical issues, exercise, pain, and social support") questionnaire, contained 4 underlying dimensions: medical issues, exercise, pain, and social support, including 9, 4, 7, and 4 questions, respectively. The content of the questionnaire was judged mainly to be easily understood, relevant, and exhaustive. Intraclass correlation coefficients for the test-retest reliability of the questionnaire answers over a week ranged from 0.68 to 0.96 for single questions. CONCLUSION: The MEPS questionnaire appears to be a valid and reliable tool for assessing the perceived ability to manage JCA. Whether it is sufficiently responsive to interventions remains to be investigated. PMID- 10689988 TI - Isometric strength measurements in children with arthritis: reliability and relation to function. AB - OBJECTIVE: To examine the reliability of testing strength in children with juvenile arthritis (JA), and to determine the relationship between strength and function. METHODS: Children with JA were tested for grip and pinch strength (n = 32). Isometric force produced by hip abductors and knee extensors was tested with a hand-held dynamometer (n = 29). Two therapists both performed each of the tests twice so that intrarater and interrater reliability could be examined. Function was measured by means of the Childhood Health Assessment Questionnaire (CHAQ) and performance on a 50-meter run. Reliability was examined with intraclass correlations (ICC). The relationships of strength and function were determined with Pearson and Spearman correlations. RESULTS: All measures demonstrated good intrarater (ICC = 0.92-0.97) and interrater (ICC = 0.80-0.95) reliability. Grip strength and pinch were correlated with the CHAQ (r = -0.45 and -0.33, respectively), while hip abduction and knee extension torque were correlated with rankings on the 50-meter run (rho = -0.34 and -0.38, respectively). CONCLUSION: Isometric strength can be reliably measured in children with arthritis in a clinical setting. PMID- 10689989 TI - Health-related quality of life in ankylosing spondylitis: a survey of 175 patients. AB - OBJECTIVE: To identify aspects of health-related quality of life that are most commonly affected in patients with ankylosing spondylitis (AS). METHODS: One hundred seventy-five participants in a longitudinal study of health status in AS completed a cross-sectional survey that asked them to rate the presence and importance of problems in 23 aspects of quality of life, including symptoms, disability, mood, relations with others, and concerns about treatments and the future. Participants also completed the Medical Outcomes Study Short Form 36 Health Survey (SF-36). RESULTS: The mean age of the participants was 51.1 years, and the mean duration of AS was 23.7 years; 119 (68%) were men. The most prevalent quality of life concerns included stiffness (90.2%), pain (83.1%), fatigue (62.4%), poor sleep (54.1%), concerns about appearance (50.6%), worry about the future (50.3%), and medication side effects (41%). Compared with those who had some college education, participants with 12 years of education or less were 2 to 4 times more likely to have problems or concerns with medication side effects, mobility, housework and self-care tasks, coping with illness, anxiety, payment for treatment, and relationships with spouses, family, and friends. Mean scores on the 8 domains of the SF-36 (range 0-100; higher scores indicate better function) ranged from 49 (energy/fatigue) to 77 (role limitations due to emotional problems). Patients with 12 years of education or less had significantly lower scores than those with some college on all domains except general health. CONCLUSIONS: In addition to pain and stiffness, fatigue and sleep problems are important concerns in patients with AS, while few reported problems with mood or social relationships. Less educated patients had lower quality of life in many different aspects. PMID- 10689990 TI - Systemic lupus erythematosus in three ethnic groups. IV. Factors associated with self-reported functional outcome in a large cohort study. LUMINA Study Group. Lupus in Minority Populations, Nature versus Nurture. AB - OBJECTIVE: To identify features of systemic lupus erythematosus (SLE) associated with poor functional outcome as measured by the 36-item Medical Outcomes Study Short Form 36 Health Survey (SF-36). METHODS: Two hundred twenty-four patients with early SLE (70 Hispanic, 83 African American, and 71 white) enrolled in a longitudinal study of outcomes were evaluated at study entry. The 8 composite scales and 2 summary measures (physical and mental) of the SF-36 were the dependent variables. Independent variables--1) sociodemographic, 2) clinical features, 3) immunologic, 4) global scores, and 5) behavioral/cultural--were examined for each of the scales and summary measures and for each ethnic group. Significant variables in these analyses were then used to construct models to determine their association with each of the scales and the 2 summary measures for the entire population and the 3 ethnic groups. RESULTS: Self-reported physical and mental functioning were most consistently associated with abnormal illness-related behaviors, helplessness, fatigue, and pain at study entry. Helplessness was more strongly associated with functioning in the Hispanics than in the African American or white patients. Pain was strongly associated with physical but not mental health. The models were quite robust, accounting for 41% to 68% of the variance for the two summary measures. CONCLUSION: Patients' attitudes toward their disease, fatigue, and pain have greater impact on self perceived functional levels, as measured by the SF-36, than do more objective measures of disease activity and damage such as the presence of specific autoantibodies and/or the occurrence of specific organ involvement. Interventions designed to improve outcome may need to include ethnic-specific as well as general strategies. PMID- 10689991 TI - Systemic lupus erythematosus in three ethnic groups. V. Acculturation, health related attitudes and behaviors, and disease activity in Hispanic patients from the LUMINA cohort. LUMINA Study Group. Lupus in Minority Populations, Nature versus Nurture. AB - OBJECTIVE: To assess the relationship between acculturation and clinical, socioeconomic-demographic, and behavioral/psychosocial features in Hispanic patients with systemic lupus erythematosus (SLE) from the LUMINA (Lupus in Minority Populations, Nature versus Nurture) cohort. METHODS: An empirically derived questionnaire was administered to 67 Mexican American SLE patients participating in a longitudinal study of outcome. This questionnaire inquired about place of birth, upbringing and length of stay in the United States, language (proficiency, usage, and preferences; English/bilingual versus Spanish), type of neighborhood, self-identity, and social interactions. Responses to this questionnaire and an informal interaction with a single bilingual, bicultural Mexican American research assistant were used to generate a score on a 10-cm anchored visual analog scale (VAS) (0 = no acculturation and 10 = maximum acculturation). The responses to the questionnaire were then quantified and scored by a physician who was unaware of the VAS. A composite score was then obtained utilizing 4 of the 6 components of the instrument. The VAS was found to have adequate sensitivity (91%), specificity (88%), and overall predictive value (89%) when the composite score was used as the validity criterion. Therefore, the VAS was used in all subsequent analyses; the median in this VAS separated patients into high and low acculturation groups. The relationship between acculturation and sociodemographic, behavioral/psychosocial (social support, abnormal illness-related behaviors, and helplessness) and clinical variables (disease duration, onset type, number of American College of Rheumatology criteria met, disease activity, and damage) at study entry was then explored. RESULTS: Patients in the low acculturation group had fewer years of education, were less likely to have private health insurance, and had less social support as compared with those in the high acculturation group; they also exhibited less disease activity as determined by the overall physician and patient global assessments of the Systemic Lupus Activity Measure. Abnormal illness-related behaviors and helplessness were not increased in the low acculturation group. CONCLUSIONS: Low levels of acculturation were associated with indicators of low socioeconomic status, but also with less disease activity at enrollment into LUMINA; they were, however, not associated with more abnormal illness-related behaviors or with helplessness, as measured in this study. The possible impact of acculturation and of its mediators in the course and outcome of SLE among Hispanic patients needs to be determined longitudinally. PMID- 10689992 TI - Determinants of shoulder and elbow flexion range: results from the San Antonio Longitudinal Study of Aging. AB - OBJECTIVE: To gain a knowledge of factors associated with impaired upper extremity range of motion (ROM) in order to understand pathways that lead to disability. METHODS: Shoulder and elbow flexion range was measured in a cohort of 695 community-dwelling subjects aged 65 to 74 years. Associations between subjects' shoulder and elbow flexion ranges and their demographic and anthropometric characteristics, as well as the presence of diabetes mellitus or self-reported physician-diagnosed arthritis, were examined using multivariate regression models. The relationship between shoulder or elbow flexion range and subjects' functional reach was examined to explore the functional significance of ROM in these joints. RESULTS: The flexion range for the 4 joints studied was at least 120 degrees in nearly all subjects (> or = 99% of the subjects for each of the 4 joints). Multivariate models revealed significant associations between male sex, Mexican American ethnic background, the use of oral hypoglycemic drugs or insulin to treat diabetes mellitus, and a lower shoulder flexion range. A lower elbow flexion range was associated with male sex, increasing body mass index, and the use of oral hypoglycemic drugs or insulin. A higher shoulder or elbow flexion range was associated with a lower likelihood of having a short functional reach. CONCLUSIONS: The great majority of community-dwelling elderly have a flexion range of shoulder and elbow joints that can be considered functional. Diabetes mellitus and obesity are two potentially treatable factors associated with reduced flexion range of these two functionally important joints. PMID- 10689993 TI - Translation to Spanish, reproducibility, and cross-cultural adaptation of the Miller-Rahe Recent Life Change Questionnaire in Venezuela. AB - OBJECTIVE: To translate into Spanish a version of the Miller-Rahe Recent Life Change Questionnaire and to adapt it to Venezuelan cultural values. METHODS: The Spanish version and cross-cultural adaptation of the Miller-Rahe Recent Life Change Questionnaire was done following recently proposed guidelines to preserve semantic, idiomatic, and conceptual equivalence in translations of health assessment instruments. We performed one or more translations into the new language, as well as back-translation, test-retest reliability, and weighting score for the translated instrument. RESULTS: A Spanish version of the Recent Life Change Questionnaire was obtained. Validity of translations was demonstrated with a significant agreement for the nonliteral translations (kappa value = 0.97, P < 0.05). The conceptual equivalence was demonstrated by significant agreement in the back-translations (kappa value = 0.84, P < 0.05). The translated instrument met acceptable levels of reliability, as assessed by Spearman's rank correlation coefficients (> 0.60 for all categories of the questionnaire). The cross-cultural adaptation of the translated instrument required addition and exclusion of items as well as changes of the ranking and scaling of life units in the original questionnaire. CONCLUSION: A valid and reliable Spanish version of the Miller-Rahe Recent Life Change Questionnaire was produced and adapted to Venezuelan cultural values. PMID- 10689994 TI - A history of early investigation in polyarteritis nodosa. PMID- 10689995 TI - Neurologic symptoms in simple neck sprain: comment on the review by Ferrari and Russell. PMID- 10689996 TI - Building nursing's future. Expanding AWHONN's expertise. PMID- 10689997 TI - Stopping neural tube defects. PMID- 10689998 TI - Genetics and discrimination. How nurses can help women make smart choices. PMID- 10689999 TI - Diagnosis infertility. PMID- 10690000 TI - The business of women's health. Building a successful women's health practice. Part Two. PMID- 10690001 TI - Building blocks. How one hospital designed the core components of a new NICU. PMID- 10690002 TI - Cardiopulmonary resuscitation in pregnancy. What all nurses caring for childbearing women need to know. PMID- 10690003 TI - Evidence! Show me the evidence! Untangling the web of evidence-based health care. PMID- 10690004 TI - Managing your career. Staying flexible in an evolving nursing industry. PMID- 10690005 TI - Dealing with difficult people. Tips and techniques for enhancing communication. PMID- 10690006 TI - Preventing neural tube defects: my personal story and mission. PMID- 10690007 TI - What's all the fuss about "older women?". PMID- 10690008 TI - Sign on the dotted line. Exploring your professional contract. PMID- 10690009 TI - Balancing the scales. Nutritional counseling for women with eating disorders. PMID- 10690010 TI - Charting in record time. Setting guidelines for documenting FHR enhancing care for laboring women. PMID- 10690011 TI - Postpartum depression. Stopping the thief that steals motherhood. PMID- 10690012 TI - Continuing education gets wired. PMID- 10690013 TI - Capitalizing on nursing creativity. PMID- 10690014 TI - Positive outcomes. Sharing good news in a caring way. PMID- 10690015 TI - So what's the scoop about the certificate program in holistic nursing? PMID- 10690016 TI - Dharma and the open heart. PMID- 10690017 TI - President's report of the State of the Association. PMID- 10690018 TI - A closer look at the consumer bill of rights and responsibilities. PMID- 10690019 TI - Reclaiming your own creativity. PMID- 10690020 TI - Springtime. Awakening of holism in our lives. PMID- 10690021 TI - Fragrances: friend or foe? PMID- 10690022 TI - Program report for nurses certificate program in Interactive Imagery. PMID- 10690023 TI - Watsu-aquatic bodywork. PMID- 10690024 TI - The crystal within. PMID- 10690025 TI - An open letter to my friends and colleagues at AHNA. From the Holistic Nurse of the Year, 1998. PMID- 10690026 TI - The way changes are made: incrementalism. PMID- 10690027 TI - Healing touch certificate program. PMID- 10690028 TI - What makes research holistic? PMID- 10690029 TI - Tri-level affirmations: the key. PMID- 10690030 TI - Insurance reimbursement. PMID- 10690031 TI - The Nightingale moment. Interview by Marie Fasano-Ramos. PMID- 10690032 TI - Dolphins and a path of healing. PMID- 10690033 TI - Holistic healing: heritage to vision. Report of the 1999 Annual Conference Scottsdale, Arizona. PMID- 10690034 TI - A new healing path PMID- 10690035 TI - Creating environmentally friendly spaces--Part I: Detoxification oasis. PMID- 10690036 TI - Building bridges--a holistic model for practice. PMID- 10690037 TI - Think this can't happen to you? PMID- 10690038 TI - Readers question whether drug increases side effects of chemotherapy. PMID- 10690039 TI - Leukocyte reduction filters may cause hypotension. PMID- 10690040 TI - A critical pathway for patients undergoing one-day breast cancer surgery. AB - As the trend of surgical procedures shifting from inpatient to outpatient settings continues, outpatient-focused standardized care processes will become more of a necessity. A multidisciplinary critical pathway (CP) for breast cancer surgery can assist care providers in meeting patients' educational and psychosocial needs. The CP document discussed in this article takes into account the expedient nature of outpatient surgery and spans the continuum of care from the surgical clinic to the postoperative homecare visit. Integrating homecare nursing improves the quality and consistency of care. PMID- 10690041 TI - A review of Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia (WM) is an uncommon B cell malignancy that resembles other B cell malignancies, such as multiple myeloma and chronic lymphocytic leukemia. WM's clinical course varies widely, with survival ranging from 1-16 years. WM is diagnosed when a bone marrow biopsy reveals malignant B lymphocytes arrested at the plasmacytoid lymphocytic stage of the maturation process and when high levels of IgM are found in the serum blood. Common clinical features include anemia, abnormal bleeding, and hyperviscosity, and 20%-40% of patients present with lymphadenopathy or splenomegaly. Alkylating agents have been the most common chemotherapy agents used to treat WM. However, nucleoside analogues are being used more frequently with promising results. Nursing care includes educating the patient about the disease trajectory, providing symptom management, and monitoring the patient's response to treatment. PMID- 10690042 TI - Closing the gap in prophylactic antiemetic therapy: patient factors in calculating the emetogenic potential of chemotherapy. AB - The ability to provoke emesis is defined by the emetogenic potential of each antineoplastic agent and by individual prognostic factors that determine the risk for each patient. The risk of chemotherapy-induced emesis is increased for females, patients between the ages of 6 and 50, and patients who drink little or no alcohol. Other risk factors include susceptibility to motion sickness and high levels of anxiety. Patients with one or more risk factors may require antiemetic treatment typically prescribed for a highly emetogenic regimen, even when a chemotherapy regimen is considered moderately emetogenic. The 5-HT3 receptor antagonists are the most effective agents against chemotherapy-induced nausea and should become standard antiemetic therapy for high-risk patients. Knowledge of factors affecting emesis and the antiemetic agents available for treating high risk patients are the keys to successful nursing management of emesis in patients receiving chemotherapy. PMID- 10690043 TI - Discrimination in the workplace. PMID- 10690044 TI - Advances in symptom management: lymphedema. PMID- 10690045 TI - Disseminated intravascular coagulation. PMID- 10690046 TI - Amifostine. PMID- 10690047 TI - Picture this. Oral candidiasis. PMID- 10690048 TI - Dyspnea and palliative care. PMID- 10690049 TI - Poster and podium sessions provide insight into various oncologic side effects and treatments. PMID- 10690050 TI - Prescriptive privileges, authority, or responsibility? PMID- 10690051 TI - Two reports of expert practice: nursing matters. PMID- 10690052 TI - Changing pain management practice and impacting on patient outcomes. AB - Concerns about acute pain management prompted the development of a pain management program. The program, with assessment, intervention, and evaluation components, was introduced by the clinical nurse specialist (CNS) in collaboration with a multidisciplinary team. The assessment phase included a descriptive study of postoperative pain and a baseline audit of nursing documentation related to pain assessment and management. Interventions, including a focused educational program and new routes for postoperative analgesia, were then introduced. Evaluations at 3 months and 2 years indicated improvements in documentation of pain assessments and improved management of pain. Additionally, at each follow-up evaluation, patients reported decreased postoperative pain. The assessment and management of pain is an important domain of nursing practice. The CNS is in a unique position to influence nursing practice and to impact on patient outcomes in this area of pain management. PMID- 10690053 TI - The price of medical progress: the advance directive. PMID- 10690054 TI - Case management of the frail elderly in the community. AB - Nurse case management has become a popular strategy for coordinating healthcare services to high-risk populations. This article describes the characteristics and advantages of a unique case management approach to manage the healthcare of frail elderly living in the community. At the heart of this approach are the nurse's role in the engagement of the client and family, prevention, continuity of care, and hospital, community, and caregiver team facilitator. Specific interventions of the nurse case manager are highlighted by case studies. PMID- 10690055 TI - So where are we? PMID- 10690056 TI - A description of the roles, activities, and skills of clinical nurse specialists in the United States. AB - Clinical nurse specialists (CNSs) frequently adapt to meet the challenging and changing needs of patients, families, nurses, physicians, and institutions, thus creating an advance practice role that is problematic in definition and description. The two dilemmas associated with CNSs have been role confusion and ambiguity, and the inability to explicate CNSs' value in economic terms. The purpose of this study was to describe the roles, activities, skills, and the cost saving and revenue-generating activities of Master's-prepared nurses who function in traditional CNS roles in the United States. A descriptive research design was employed, using Role Theory as a framework to guide the study. The tool used to measure CNS practice included a 68-item instrument. It was pretested and used in two pilot studies. Content validity was supported by three experienced CNSs who were, at the time, in a doctoral nursing program. Instrument reliability was 0.89. Surveys were mailed to all individuals who subscribed (n = 2379) to the Clinical Nurse Specialist Journal. From the convenience sample, 724 CNSs participated, providing a margin of error of +/- 4 percentage points with a 99% confidence level. Regarding the five role components, CNSs reported (listed from most frequently to least frequently) spending time in the role of expert practitioner, educator, consultant, administrator, and researcher. Most of the activities listed in each of the roles were typical of CNS practice. Of the advanced practice roles, the two with the most surprising results were the expert practitioner and administrator roles. The results indicated a trend toward performing advanced skills that have been in the past considered solely medical practice and toward increasing administrative responsibilities. A small number of CNSs were able to identify cost-saving and revenue-generating activities, including the monetary value of the activity. PMID- 10690057 TI - Political influence: a model for advanced nursing education. AB - Involvement in shaping healthcare policy is an expected outcome for the leadership standard of advanced nursing practice as outlined by American Nurses Association's Scope and Standards of Advanced Practice Registered Nursing (1996). From personal experience teaching a healthcare policy course over a 10-year period, the author has found that beginning graduate students are primarily in an apolitical state, reluctant to participate in such activity, and have not considered the ramifications of inactivity for the profession and for healthcare policy. The skills for political influence are an essential component of graduate education to increase awareness of, and foster activity in, the political arena. A model for the development of these skills is presented. The model consists of four interrelated dimensions: information, commitment, initiative, and involvement. Model dimensions are described and made operational through student activity. Ultimately, students move from an apolitical state to a moderate level of involvement. The model has been intended for educational purposes; however, it is applicable for any advanced practice nurse (APN). PMID- 10690058 TI - Strategies for working with a state board of nursing. PMID- 10690059 TI - Is Louisiana a reflection on the state of master's-prepared nurses nationally? PMID- 10690060 TI - Hospital and parish (county) utilization of master's-prepared nurses in Louisiana. AB - Recent regulation in Louisiana involving advanced practice nursing, combined with new educational standards at the Master's level established by the Louisiana State Board of Nursing, prompted a statewide survey of hospitals and parish health units. The purpose of the survey was to determine whether and to what extent Master's-prepared nurses were being used in Louisiana and to identify the importance placed on role functions and activities generally associated with the advanced practice nurse (APN). The survey included open-ended questions concerning expectations about future hirings and perceptions of future roles and duties of the Master's-prepared nurse as well as suggestions to educators on what is needed to better prepare nursing students at the Master's level. Results from this exploratory survey provide information on APN utilization with implications for nursing educators and administrators as well as APNs. PMID- 10690061 TI - Creating "asset-rich" environments. PMID- 10690062 TI - Partners in psychiatric-mental health nursing. PMID- 10690063 TI - Homeless parents: parenting education to prevent abusive behaviors. AB - PROBLEM: This study sought to measure the effects of a short-term parenting course for homeless parents to decrease their parenting stress and potential for abusive parenting behaviors. SAMPLE: Nineteen homeless parents lived in a family emergency shelter for homeless families in Hillsborough County, FL. Their average length of shelter stay was 3 weeks. METHODS: Residents were invited to participate in a parenting education course consisting of three 1-hour classes weekly for 3 consecutive weeks. Class content focused on the parent, the child, and discipline. Subjects' potential for child abuse was measured with the Adult Adolescent Parenting Inventory and their parenting stress with the Parenting Stress Index. A quasi-experimental pre/posttest design was used, with a t test statistic at the .05 level of significance. Descriptive statistics were used to analyze selected demographic data. FINDINGS: Results revealed significant differences in the scores of parenting stress originating from the child domain of the PSI and in the scores on the construct of unrealistic expectations of the child in the AAPI. Both scores had decreased on the posttest. CONCLUSIONS: Short term parenting education courses may be a useful intervention strategy. PMID- 10690064 TI - Parental murder and suicide: post-traumatic stress disorder in children. AB - TOPIC: Post-traumatic stress disorder (PTSD) diagnosed in a 9-year-old who witnessed her parents' murder and suicide. PURPOSE: To provide a historical overview of PTSD in children in general and to discuss one case study in particular. SOURCES: Personal observations and experiences, and a review of both classic and contemporary literature in child psychiatry and developmental psychology. CONCLUSIONS: Supportive psychotherapy for a traumatized child with clinical supervision for the care-giver showed evidence of the recovery process in one PTSD case study. PMID- 10690065 TI - Beyond cause and effect: some thoughts on research and practice in child psychiatric nursing. AB - TOPIC: A comprehensive conceptual framework that informs about the complexity of childhood psychopathology. PURPOSE: To discuss the importance of a unifying conceptual framework and multiple partnerships in research and practice that can inform critical events and effects of interventions at various system levels in the treatment of children. SOURCES: Literature review. CONCLUSIONS: The most informative research and intervention will occur by using multiple measures and sources of information. As understanding of children and their problems in development and in context grows, intervention research that will be both useful, timely, and cost-effective must include cross-discipline teams of researchers and practitioners who will speak to these complexities. PMID- 10690066 TI - Kaleidoscope of excellence. An interview with Judy Coucouvanis, MA, RN, CS, NP. Interview by Linda M Finke. PMID- 10690067 TI - The therapeutic use of music for dyspnea and anxiety in patients with COPD who live at home. AB - The purposes of this repeated measures study were to examine the feasibility of using music as an intervention for dyspnea and anxiety in patients with chronic obstructive pulmonary disease (COPD) who live in their homes and to examine the effect of music on anxiety and dyspnea. Twenty-four participants who experienced dyspnea at least once a week were studied over a 5-week period. Baseline data were collected on Week 1. Measures of anxiety and dyspnea were taken on Week 2, prior to and immediately following the use of music. These measures were repeated on Week 5. There was a significant decrease in dyspnea following the use of music as reported in the music diary (p < .001). There was a significant decline in anxiety (p < .05) and dyspnea (p < .01) following the use of music on Week 2. There was no significant change in anxiety or dyspnea over the 5-week period. PMID- 10690068 TI - Self-transcendence and activities of daily living. The woman with the pink slippers. AB - The number of older adults in our population is steadily increasing. Many older adults continue to remain active and care for themselves. However, differences exist in older adults' ability to perform activities of daily living. The purpose of the study was to explore relationships among self-transcendence (ST), health status (SHS), and ability to perform activities of daily living (ADL) in noninstitutionalized older adults. The 88 participants were primarily widowed, White women, 65 years of age and older (M = 73.4), who perceived their health positively, and had 12 years or more of education. Findings included statistically significant relationships between ST and ADL and SHS and ADL. Twenty-two percent of the variance in ability to perform ADL was explained by SHS, and an additional 6% was explained by ST. Nurses are encouraged to explore factors that contribute to older adults' ability to remain independent. PMID- 10690069 TI - A research-based use of Tai Chi/movement therapy as a nursing intervention. AB - Tai Chi has been widely practiced in China for centuries as an art form, religious ritual, relaxation technique, exercise, and a method of self-defense for people of all ages. It has been used to improve balance; promote postural stability; decrease falls; enhance cardiovascular and ventilatory functions; rehabilitate persons with acute myocardial infarction and rheumatoid arthritis; and reduce pain, stress, and nightmares. The purpose of this article is to summarize, synthesize, and critically evaluate the research-based use of Tai Chi presented in the current literature and give implications and directions for future research. Additional studies about the effects of Tai Chi from a nursing perspective are needed to make clear when it is beneficial as a nursing intervention. PMID- 10690070 TI - Cultural and spiritual meanings of childbirth. Orthodox Jewish and Mormon women. AB - This descriptive, phenomenological study investigated the cultural and spiritual meanings of the childbirth experience from the personal perspectives of 30 Canadian Orthodox Jewish and 30 American Mormon women. Fewer Jewish women had childbirth education and attendance of their partners during childbirth than did Mormon women. Participants in the study, having codified belief systems, expressed the primary importance of bearing children in obedience to religious law. Birth was articulated as a bittersweet paradox, often accompanied by a sense of empowerment. Women described the importance of personal connectedness with others and with God, the importance of childbearing, and the spiritual and emotional dimensions of their childbirth experiences. Religious beliefs help women define the meaning of childbirth and may provide coping mechanisms for the intensity of giving birth. It is essential for holistic nurses to value and acknowledge the cultural and spiritual dimensions of the childbirth experience. PMID- 10690071 TI - A collaborative research project on Therapeutic Touch. AB - The current shift toward granting funds for collaborative research proposals means that graduate nursing students must be prepared to participate in the collaborative research process. The authors describe how master's and doctoral students worked together with faculty to establish group norms, investigate, design, and disseminate a research proposal on Therapeutic Touch. Project goals, description of group members, and the benefits and limitations of collaborative research also are included. Evaluating the collaboration as a challenging yet enjoyable learning experience, students and faculty shared a successful scholarly endeavor that ultimately led to funding and implementation of a research proposal on Therapeutic Touch. PMID- 10690072 TI - Shared governance: time to consider the cons as well as the pros. AB - AIMS: This paper aims to provide a critical appraisal of an approach to the management and organization of nursing work known as shared governance (SG). BACKGROUND: This approach has its origins in the USA, where, during the past 20 years it has become increasingly influential. The advocates of SG claim that it can, inter alia, improve recruitment and retention rates, boost morale, and help raise clinical skills. Little wonder that SG in now beginning to make significant inroads into the NHS. ORIGIN OF INFORMATION: However, a trawl through the extensive US literature, using printed and online (e.g. BIDS, CINHAL, MEDLINE, etc.) bibliographical sources, suggests that the claimed benefits of SG should be treated with caution. KEY ISSUES: Much of the existing published research appears to be both methodologically flawed and lacking in any critical edge. While many researchers and commentators appear only too willing to highlight what they see as the promise of SG, they shy away from exploring any potential pitfalls. One consequence of this is that many of the putative benefits SG is said to confer, may in fact be more apparent than real. CONCLUSIONS: Nurses and nurse managers need to be apprised of and consider seriously, the possible cons as well as the potential pros of SG, if any promise it may have is to be realized. PMID- 10690073 TI - Managing the unmanageable: risk assessment and risk management in contemporary professional practice. AB - AIM: This study sets out to investigate the theories and practices of risk assessment and management in the context of contemporary mental health practice. BACKGROUND: Although risk assessment and management policies are well established for those working in the field of community mental health care, there are noticeable anomalies and regional variations, in the criteria, procedures and decision-making strategies used. METHODS: Focus group taped interviews were conducted with over 100 mental health professionals in one NHS Trust. These were compared with an extensive literature review on the topic. FINDINGS: The main theme to emerge was lack of resources, which included time and staff in the context of a changing and increasing workload. Another important theme was the lack of access to centralized and accurate information about mental health service provision. CONCLUSION: It is essential that professionals, clients, their families and the public feel confident in professional judgements and practices to avoid a 'back to the asylum' lobby, for the care and treatment of seriously mentally ill individuals. PMID- 10690074 TI - Job satisfaction and stress in staff working in a specialist psychiatric unit for the elderly following relocation from a traditional psychiatric hospital setting. AB - AIM: This study examines self-reported stress and job satisfaction of staff working in a specialist psychiatric unit for the elderly (n = 79) following relocation from a traditional psychiatric hospital setting (n = 66). The levels of reported stress were examined in relation to the two staff groups before and after the relocation, and in relation to data available for similar occupational groups. METHODS: Stress and job satisfaction levels were surveyed using the Occupational Stress Indicator. The analysis of data included t-test for independent groups. FINDINGS: The results suggested that that there were no significant changes in the patterns of the staff groups' experience of stress at the traditional psychiatric hospital and later at the new purpose-built unit. However, it was felt that there may have been different factors contributing to staff stress at the two time-points. The staff groups in this survey reported similar, and in some areas more positive levels of occupational stress than both community and other health authority samples. However, it was felt that, due to the limited response rate of staff at both times (63 and 59%), the results may be an underestimate of stress levels. In the absence of a measure of general health and symptomatology it was not possible to assess the impact of stress levels on the psychological well-being of staff. CONCLUSIONS: Routine audit of staff stress would be beneficial to identify potential for staff training, and individual professional development plans. This is particularly important in view of the current difficulties both locally and nationally in the recruitment and retention of qualified staff in this speciality. PMID- 10690075 TI - Quality in post-basic nurse education: the need for evidence-based provision. AB - BACKGROUND: As one means of addressing the problem of providing high quality health care within a context of diminishing resources, the British National Health Service (NHS) has promoted the concept of evidence-based clinical care. In order to integrate this concept effectively within routine practice, nurses need a sound knowledge of fundamental research methods. Unfortunately, the research skills courses that have been provided have typically relied on assumption to determine course content and chance factors for recruitment. Unsurprisingly, such haphazard provision is highly wasteful of resources, targeting neither the real skill deficits of the workforce nor the personnel most in need of this type of training. AIM: To demonstrate a more systematic approach to identifying the skill deficits of any specified workforce. METHODS: A survey was conducted with a random sample of nurses, who covered a range of grades and clinical specialities, using a psychometrically valid and reliable training needs analysis tool. FINDINGS: The results revealed both common training needs as well as skill deficits pertinent to a given locality and clinical area. CONCLUSIONS: Using more objectively derived information affords the commissioning of customized research skills courses that have the capacity to meet the needs of both the local organization and its employees. In this way, limited training budgets can be more effectively targeted. PMID- 10690076 TI - Preparing adult branch students for their management role as staff nurses: an action research project. AB - AIM: This action research project aimed to investigate students', newly qualified staff nurses' and ward managers' views about the management skills and knowledge required by staff nurses, and how best students could be prepared for their management role. BACKGROUND: The importance of the staff nurse's management role is increasingly being recognized but the literature highlights the difficulties in preparing students for management, and personal experience confirmed this. METHODS: Focus group interviews were held with senior students and newly qualified staff nurses. A questionnaire was then developed which was completed by 23 ward managers. FINDINGS: A good insight into the management skills and knowledge expected of newly qualified staff nurses, and useful ideas about preparation for the role emerged. An extra 2 weeks in practice, supernumerary and with specific management learning outcomes, was implemented and evaluated well. CONCLUSIONS: Preparation of students for their management role as staff nurses benefits from being closely linked to practical experience, with clear learning outcomes and supportive clinical staff. Using an action research approach to investigate the problem and develop a strategy was found to be an appropriate methodology. PMID- 10690077 TI - Outcomes of implementing primary nursing in the care of people with chronic lung diseases: the nurses' experience. AB - AIM: This study describes the outcomes of an action research project carried out to implement primary nursing in the care of people with chronic lung diseases at two hospital units in Iceland. METHODS: The methodological approach was the interpretative perspective. Data from transcribed interviews with 21 nurses and a research journal written by the author were analysed. Themes were generated according to dialectical procedures of interpretation. FINDINGS: The following themes were identified: close relationships with patients, continuity of care, reports of satisfied and secure patients, centrality of individual patient's needs, constant refinement of the system, sensitivity to staffing load and ambitious and responsible nurses. CONCLUSIONS: The nurses participating in this study clearly valued the possibilities that primary nursing brought in terms of higher quality of care for their patients. One of the most important components of high quality care is to know and understand patients' experiences, which is the main outcome of this study. This indicates that implementing primary nursing in the care of people with chronic lung diseases was beneficial from the point of view of the nurses. However, concerns were raised that primary nursing is more sensitive to low staffing than the system it was meant to replace. PMID- 10690078 TI - Helping families make time. PMID- 10690079 TI - What is new in asthma: new dry powder inhalers. AB - Asthma affects an estimated 4.8 million children. The pressurized metered-dose inhaler (pMDI), despite problems associated with its use and concern that most of these inhalers contain ozone-damaging chlorofluorocarbons (CFCs), is currently the device most frequently used to deliver inhaled medication. Concerns regarding pMDIs that contain CFCs have led to further development of alternative delivery devices, including CFC-free pMDIs, breath-actuated devices, and dry powder inhalers (DPIs). Advantages and disadvantages of these devices are discussed briefly, with emphasis on the new DPIs. A brief overview of their safety, efficacy, and acceptance by patients is presented. DPIs have the potential to become important devices for administration of inhaled medication in pediatric asthma management. PMID- 10690080 TI - The President Clinton crisis and the Starr report: children's perceptions and parents' awareness. AB - INTRODUCTION: The media have suggested that the President Clinton crisis, as publicized by the Starr Report, has had detrimental effects on school-age children. Parents, too, have been concerned that their children are confused about the presidential controversies involving dishonesty, mistrust, betrayal, infidelity, and misuse of authority. The purpose of this study was to explore the perceptions and reactions of school-age children to information surrounding the President Clinton situation, as well as parents' perceptions of their children's knowledge and reactions. METHOD: A descriptive, qualitative design was used in this study. Data collection took place during the 2-week period following publication of the 1998 Starr Report. Fifty-one parents and 67 school-age children were interviewed using semi-structured interview guides with 5 open ended questions addressing the President Clinton situation. RESULTS: Major themes that emerged from children's interviews were lying, getting caught, infidelity, and role modeling. Parents' awareness of their children's knowledge regarding the President Clinton situation varied; few were aware of the depth of knowledge their children had, nor had they discussed this situation with their children. DISCUSSION: Findings have relevance for nurse practitioners as they support parents in listening to and talking with their children about tough issues such as morality and sexuality in the context of real life events. PMID- 10690081 TI - Infant cranial molding deformation and sleep position: implications for primary care. AB - Infant positional plagiocephaly, a cranial molding deformity expressed by asymmetrical head shape, is on the rise. The increase correlates with the recommendation by the American Academy of Pediatrics that infants be placed on their backs to sleep to reduce the risk of sudden infant death syndrome. The majority of misshapen heads develop because infants are placed to sleep in the same supine position, without head rotation. Infant molding deformities will generally improve with repositioning and cranial growth, but permanent deformation can occur, especially without early treatment. This article informs pediatric practitioners about positional plagiocephaly, offers preventative and treatment interventions, and reviews treatment options. PMID- 10690082 TI - A descriptive study of missed appointments: families' perceptions of barriers to care. AB - INTRODUCTION: When clinic appointments are missed, families deprive their child of health care opportunities and contribute to rising health care costs. The purpose of this study was to determine families' perceptions of barriers to attending clinic appointments. METHOD: Two hundred participants with a history of missed appointments were randomly selected to participate in a telephone survey. Ninety-five of the families selected did not have telephones. Of the 105 families contacted by telephone, 101 consented to participate in a survey. For the group without phones (n = 95), demographic information was collected to use as comparison data with families that were reached. Patterns of missed appointments were also analyzed. RESULTS: The majority of families were headed by young single mothers. The families identified transportation problems, wait times, and not knowing the reason for the appointment as barriers. DISCUSSION: Clearly, many issues have an impact on decisions related to attending clinic appointments. In the interest of health, interventions to decrease barriers and increase attendance should be a priority for health care professionals. Further research of the effectiveness of these interventions will delineate the appropriate focus of health care professional's efforts. PMID- 10690083 TI - New challenges, new answers: pediatric nurse practitioners and the care of adolescents. AB - INTRODUCTION: Since the 1950s, patterns of morbidity and mortality among adolescents have shifted to social and environmental causes. This study examines pediatric nurse practitioners' (PNPs') self-assessed competencies in addressing the common health concerns of adolescents. METHOD: The analysis used a sample of 257 PNPs drawn from a larger national data set of 637 nurses randomly sampled from 3 nursing organizations. Factors associated with self-perceived knowledge or skill and interest in training for 28 common health concerns of adolescents were analyzed using Chi square, t test, and Pearson's correlation. Barriers and attractions to working with adolescents were also investigated. RESULTS: The greatest deficits in self-perceived knowledge or skill, as well as low interest in training and low perceived relevance to practice, were around issues of gangs, gay/lesbian/bisexual/transgender youth, HIV/AIDS, and counseling about a positive pregnancy test. Also, PNPs identified the lack of resources appropriate for adolescent referrals as the greatest barrier to working with this population. DISCUSSION: PNPs assessed their lowest competencies in some of the areas that present the greatest threats to adolescents' health and well-being. These deficits suggest needed curricular shifts in entry-level and advanced-level preparation of PNPs, as well as new priorities for continuing education. PMID- 10690084 TI - Palivizumab for respiratory syncytial virus prophylaxis. AB - Palivizumab, a humanized monoclonal antibody, has been approved by the FDA to prevent severe lower respiratory tract infections caused by RSV in high-risk patients. Prophylaxis of RSV infections with palivizumab requires monthly injections (15 mg/kg) during the RSV season. In the IMpact-RSV study, hospitalizations resulting from RSV decreased by 55% in the palivizumab treatment group. Palivizumab has also been shown to decrease the number of days with moderate or severe RSV infection, with an increased oxygen requirement, and ICU admissions. Palivizumab has been shown to be well tolerated with minimal adverse effects. The most frequently reported adverse effects were fever and minor injection site reactions. Determination of which patients should receive RSV prophylaxis should take into consideration all risk factors. Recommendations for RSV prophylaxis with RSV-IGIV and palivizumab have been published by the American Academy of Pediatrics. To date, no studies directly comparing RSV-IGIV and palivuzumab have been conducted. Neither product is recommended in children with congenital heart disease. PMID- 10690085 TI - Girl with a recurrent cough. PMID- 10690087 TI - Disease management: what does it mean for nurse practitioners? PMID- 10690086 TI - NAPNAP supports new children's health bill. PMID- 10690088 TI - Selective serotonin reuptake inhibitors: implications for advanced nursing practice. AB - The efficacy of SSRIs in treating depression as compared to other classes of antidepressants is well documented. There is a growing body of knowledge documenting differences and similarities among these drugs, including half-life elimination, side effects, drug interactions, dosing and cost. These data must be reviewed before prescribing. Providers must take caution when giving SSRIs to patients who have been on other antipsychotics, especially MAO inhibitors, elderly patients, patients who are pregnant, and patients on type 1C antiarrhythmia agents. PMID- 10690089 TI - Abdominal aortic aneurysm: diagnosis, treatment, and implications for advanced practice nursing. AB - The nurse practitioner needs to acknowledge that not all patients are insightful. The patient who is naive, stoical, or in denial may not return to his previous level of health. Behavioral or environmental changes may be a necessary part of recovery. The advanced practice nurse, through her research and subsequent knowledge, can identify and implement holistic changes necessary for the maintenance of health and the development of appropriate health-seeking behaviors that lower the morbidity and mortality for such conditions as abdominal aortic aneurysms. Nurse practitioners play a vital role in research, prevention and early detection of major threats to wellness (Lawler and Schmidt, 1992). Gender sensitive research regarding factors affecting recovery are also necessary as females respond differently to such conditions as renal failure (Carlson and Eisenstat, 1995). PMID- 10690090 TI - Medical chemical sensitivities: an overview. PMID- 10690091 TI - Hepatitis C. AB - Hepatitis C virus infection is responsible for significant morbidity and mortality worldwide. Advances in detection and monitoring of hepatitis C virus infection, as well as treatment protocols, have contributed to the medical focus on this high profile disease. Presence of risk factors should increase the clinicians index of suspicion for this symptomatically nonspecific disease. PMID- 10690092 TI - Missed immunization opportunities: a comparison of nurse practitioners and physicians. PMID- 10690093 TI - Giardia lamblia in adults: a case study. PMID- 10690094 TI - Priori outcomes: oxymoron or practice plan. AB - This assessment helped shape the practice of a nurse administered mobile clinic serving three rural communities. Needed services were identified and existing services were customized to improve the overall health of the communities. With the dynamic state of health care systems, each practice is subject to influence by managed care and populations who, in turn, respond with changes in need, resources, and adaptation to the system dynamics. Nurse practitioners struggling to match resources and patient needs could use a similar research design to assess their practice, discover significant relationships, and redesign practice plans to fit specific practice settings. PMID- 10690095 TI - Fragile X syndrome. AB - Fragile X syndrome is the most common inherited condition causing mental retardation in males. Females with the full mutation expansion can have milder signs of the disorder. Families with members who have been diagnosed with fragile X syndrome face concerns about the health of their newborn infant, decisions regarding family planning, and questions about the possibility that other family members could have this disorder. Neonatal nurses participate in assessment, health care management, counseling, and referral of the families regarding this syndrome. PMID- 10690096 TI - Nonimmune hydrops fetalis. AB - Hydrops fetalis is a relatively rare phenomenon that presents itself in the delivery room in an extremely acute manner. Prompt resuscitation, an understanding of the condition and its presentation, and immediate treatment can make the difference between life and death in these cases. The focus of this article is on nonimmune hydrops, the type of hydrops seen in the clinical setting. Maternal and fetal pathophysiology, current theories, diagnostic evaluations, neonatal pathophysiology, clinical manifestations, treatment options, pertinent research, and the needs of the family are discussed. PMID- 10690097 TI - Percutaneously inserted polyurethane central catheters in the NICU: one unit's experience. AB - This article describes the authors' experiences with using polyurethane percutaneously inserted central catheters from June 1993, when these catheters were introduced in the NICU at Children's Hospital, Omaha, Nebraska, through September 1997. Indications for line placement and anatomy are reviewed. Patient demographics, success rates, and complications are analyzed. Line cares, including dressing changes, management of infusions, and troubleshooting, are also discussed. PMID- 10690098 TI - Efficacy of saline vs heparin in maintaining 24-gauge intermittent intravenous catheters in a rabbit model. AB - PURPOSE: The purpose of this study was to compare saline and heparin flushes in 24-gauge i.v. catheters. DESIGN: A double-blind experiment in four recorded trials was conducted using a rabbit model. Twenty-four-gauge catheters were placed in both auricle veins, and all catheters were secured in the same manner. Ampicillin was infused into each ear at the same interval, with each ear randomized to receive either the heparin solution or the saline solution. SAMPLE: The subjects were ten white New Zealand rabbits. Data were collected on 76 catheter sites. MAIN OUTCOME VARIABLE: Patency for each catheter was measured in hours and evaluated based on presence of warmth, erythema, induration, leakage, or occlusion. RESULTS: There was no significant difference in the duration of patency of the catheters between the two groups. PMID- 10690099 TI - Use of biomagnetic therapy to encourage growth in preterm neonates. PMID- 10690100 TI - Thimerosal in vaccines: a joint statement of the American Academy of Pediatrics and the Public Health Service. PMID- 10690101 TI - Visitation patterns: parents who visit "too little". PMID- 10690102 TI - Erythromycin. PMID- 10690103 TI - Keeping score in case management. PMID- 10690104 TI - Nurse-managed wound clinic. A case study in success. AB - The wound Care Clinic at Naval Hospital Charleston is a nurse-managed ambulatory clinic that has demonstrated the successful application of nursing case management in caring for patients with chronic and complex wounds. Nursing case management is an outcomes-based system of assessment, planning, provision of nursing services, coordination of interdisciplinary efforts, education, and referral. Nursing case management has been shown, in the literature and at Naval Hospital Charleston, to be an extension of role of professional nursing practice and results in decreased costs, improved quality of care, faster wound healing times, decreased complications, and greater coordination of care between specialty disciplines. These positive results are illustrated in several case studies. Nursing case management has many implications for the successful implementation of any healthcare delivery system where decreased costs and improved quality of care are valued, and it has special benefit in the complex management of chronically ill patients. PMID- 10690105 TI - Selecting and implementing a care management system. PMID- 10690106 TI - Outcomes measurement in healthcare. New imperatives for professional nursing practice. AB - Quality of healthcare is measurable and nurses in all settings are increasingly being expected to actively participate in identifying and measuring patient outcomes. To do so requires an understanding of the tools and processes currently being used to measure the quality of healthcare in this country. This article provides an overview of current efforts in the healthcare field to develop a shared outcome language and standardized national quality indicators. The challenges professional nurses face in identifying and measuring nursing sensitive patient outcomes and the responsibility to use that data to shape nursing practice are emphasized. PMID- 10690107 TI - An experience of community. AB - This is a personal experience in case management. The current surge of case management programs across the country has made a significant impact on the lives of many patients. But, does anyone ask, "What is the impact on those who are case managing?" As we share the struggles, griefs, and pains of our patients, we should never lose sight of the fact that who we are and what we did made a difference. PMID- 10690108 TI - Developing a patient education video as a tool to case manage patients who have had strokes. PMID- 10690109 TI - Efficacy of primary care in a nursing center. AB - Nursing opportunities have expanded beyond the traditional bedside role. Nurses serve in a variety of roles such as administrators, teachers, or primary care givers in a variety of settings. The role of primary care giver is a more recent role; it involves relatively independent nursing practice with clients who have acute or chronic illnesses. Client groups may include the elderly in high rise buildings, mothers and children at schools, or homeless and low-income populations at homeless shelters. This care is often provided in a nursing center. Nursing centers are nurse-managed centers in which nurses are accountable and responsible for care of clients; they are the primary provider of care and the one most seen by clients. Case managers may be in a position to refer patients to nursing centers or to work directly with nurse practitioners in nursing centers. However, questions about the primary care provided in nursing centers must be addressed for healthcare providers, insurance companies, and patients to be confident in the efficacy of this delivery system. Is the primary care comprehensive? Is it of high quality? Is it cost effective? Is it satisfactory to clients? These and other questions about the primary care provided in nursing centers must be answered to effect political and other changes needed to fulfill the role of nursing centers envisioned by early leaders of the movement. This article addresses questions related to the efficacy of primary care provided in nursing centers by family nurse practitioners. After defining efficacy, the discussion focuses on the components identified and studied in one nursing center and includes information on opportunities for case managers to utilize nursing centers for referral and appropriate follow-up of their patients. PMID- 10690110 TI - Nursing education and practice: revisiting the reunification challenge. PMID- 10690111 TI - A comparison of pain perception of elderly African Americans and Caucasians. AB - This study explored the differences and similarities in pain perception reported by 32 elderly African Americans and 32 elderly Caucasian subjects. Using the McGill-Melzack Pain Questionnaire, the study revealed that both groups chose the word nagging most frequently to describe their pain. A 2 by 2 analysis of variance indicated a statistically significant difference between the subjects in terms of the present pain intensity (PPI) (F = 6.30, df = 1, P = .015). Pearson's Product Moment Correlation revealed a moderate correlation (r = .36, P = .01) between PPI and ethnicity. PMID- 10690112 TI - Nursing faculty's handling of academic dishonesty. PMID- 10690113 TI - Integrating multidimensional stress management into a baccalaureate nursing curriculum. AB - The mandate for self-care for holistic nurses can be satisfied with a multidimensional stress management program. This article describes a quasiexperimental pilot study that assessed the need for such a program for senior students in a baccalaureate degree nursing program. Strategies are suggested to integrate these techniques into nursing education. PMID- 10690114 TI - Collaboration between community nurses and nursing faculty using substance abuse prevention focus groups. AB - Collaboration between community nurses and nurses from a university who conducted focus groups is discussed. The focus groups explored why low-income, inner-city, white women of childbearing age did not abuse drugs. This partnership effort resulted in positive, successful outcomes for both groups of nurses and yielded culturally sensitive information that may be useful in preventing substance abuse. Methods of facilitating this collaboration and results of our joint endeavors are explored. PMID- 10690115 TI - A student-developed tool for assessing safety in schizophrenic patients. AB - Schizophrenia is a chronic, disabling disease of mind and behavior. Since some schizophrenics are prone to violence, nursing students devised a safety risk assessment tool to help health care personnel screen clients in acute care settings who may be at risk for violent behavior. The tool is accurate, quick, and user-friendly, and it enhances communication among members of the multidisciplinary health care team. The results obtained from the assessment tool guide nurses and other health care team members in implementing appropriate interventions. Future research and pilot studies are warranted to increase the reliability and validity of this tool. PMID- 10690116 TI - Connecting across the miles: interdisciplinary collaboration in the evaluation of critical thinking. AB - A pilot study in which faculty from nursing and English departments at two universities in different states shared a common evaluation tool and collaborated through e-mail to evaluate the evidence of critical thinking in writing portfolios of baccalaureate and masters' nursing students. Loxley's (1997) four processes--assessment, building, managing the process, and evaluating--are used as a framework for describing collaboration among the disciplines in the two universities. Social exchange theory was used to explain the collaboration between participants. All six professors learned a great deal from studying and scoring the writings, but they learned most from each other through their e-mail dialogue. PMID- 10690117 TI - The shortage to beat all shortages. PMID- 10690118 TI - Critical thinking. The spirit of inquiry. PMID- 10690119 TI - Organ donation and the critical care nurse. AB - CACCN supports the process of organ donation. Nurses working in critical care are in a privileged position to positively influence organ donation success; however, the process can be emotionally very difficult. Facility commitment, agency culture and medical practice are also crucial to the process of organ donation. Despite our commitment to support organ donation as an option, the critical care nurse's primary responsibility is to the potential donor and their family. Throughout the process, the critical care nurse must remain non-judgmental and supportive of the family, regardless of their decision. Ultimately, the nurse must balance organ donation with the needs of the family who is experiencing the tragic and untimely loss of a loved one. Finding this balance is never an easy task. PMID- 10690120 TI - CACCN position statement. Advance directives [practice guideline]. PMID- 10690121 TI - Sucrose as analgesia for neonates experiencing "mild" pain. AB - Health care professionals who care for neonates have few treatment options for the management of mild, sporadic painful events, such as those associated with venipuncture. A number of research studies have demonstrated sucrose to be an efficacious analgesic for mild procedural pain in neonates. The historical therapy of the "sugar nipple" has even been replaced with sucrose. A discussion of the pharmacologic principles, available research regarding dose-response relationships and implications for nursing care is presented in this article to allow the reader to consider how this adjunctive therapy may be incorporated into care of the neonate. Rather than considering sucrose as a replacement for traditional analgesics, this easily administered and seemingly safe intervention could be used as another adjunctive therapy in treating mild pain for neonates. Future directions of research may identify the precise mechanism of action that sucrose takes in the neonate, the gestational and chronological ages when sucrose is most efficacious, and the consequences of frequent or repeated dosing with term and low birth weight infants. PMID- 10690122 TI - Challenging restricted visiting policies in critical care. AB - The need for family members to visit their loved ones when they have been admitted into the critical care unit was identified in 1979 by Molter in the critical care family needs inventory (CCFNI). This need has been the centre of controversy for critical care units for many years. This article provides an overview of literature that refutes some of the rationales that have been used to restrict family visiting in the critical care unit. An overview of a liberalized (open, contract, inclusive or structured) visiting policy is discussed as an option to the restricted visiting policy. PMID- 10690123 TI - Who was Cherry Ames ... and where is she now? PMID- 10690124 TI - Introduction to sinus disease: I. Anatomy and physiology. AB - Chronic rhinosinusitis is the most common chronic illness in the United States. An understanding of the anatomy of the paranasal sinuses, their functioning in health and in disease, and the contributing factors that are critical to the pathogenesis of rhinosinusitis is essential for nurses caring for patients with this prevalent disease. This paper will provide the otorhinolaryngology (ORL) nurse with an overview of the scientific principles important in rhinosinusitis, as well as presenting a framework for the understanding of rhinosinusitis and its treatment. (This paper is the first in a series of two articles. The second part will review the diagnosis and treatment of chronic rhinosinusitis.) PMID- 10690125 TI - Patient's Bill of Rights Act 1998. AB - For nurses, patient advocacy has always been a responsibility of practice. Advocacy is rooted in the concept of individual rights. Consumers of the product of health care now have increased protection because of the Patients' Bill of Rights Act of 1998. Legislation has created a template to outline important rights and processes to assure quality health care. PMID- 10690126 TI - Writing for publication Part II--The writing process. AB - You have selected a topic, gathered resources, and identified your target audience. The next step is to begin to write and organize your ideas. Initiating the actual writing process can be intimidating, especially for a novice author. This portion of the writing for publication series focuses on helping the writer to organize ideas and get started. PMID- 10690127 TI - Measuring sedation in the ICU: guidelines on the scales? PMID- 10690128 TI - Orexins: a new family of neuropeptides. PMID- 10690129 TI - The Brussels sedation scale: use of a simple clinical sedation scale can avoid excessive sedation in patients undergoing mechanical ventilation in the intensive care unit. AB - Sedation is an important component of patient comfort in the intensive care unit (ICU), especially in those undergoing mechanical ventilation. Sedation that is too light or too deep can have important consequences, and therefore assessment of the degree of sedation should be an important part of patient management. Although there are many methods available to assess the degree of sedation, none is ideal. Therefore, we developed a new sedation scale and analysed its clinical impact in the management of patients undergoing mechanical ventilation. The study comprised two consecutive phases. In the first phase, the medical team did not use a sedation scale. In the second phase, the medical staff used the new sedation scale, comprising five levels, depending on the perceived degree of sedation: levels 1 and 2 = oversedation; levels 3 and 4 = correct sedation; and level 5 = undersedation. There were no significant differences in mean or highest levels between patients in the two phases (mean 2.89 (SD 0.11) vs 2.67 (0.13), P = 0.22; highest 3.16 (0.11) vs 3.10 (0.14), P = 0.78). However, the lowest level was significantly greater in patients in the second phase than in those in the first phase (2.61 (0.11) vs 2.16 (0.13); P = 0.011), indicating that the number of patients with excessive sedation was significantly reduced with the introduction of this scale. Thus the use of this scale can have a real clinical impact for patients undergoing mechanical ventilation, principally by avoiding excessive sedation. PMID- 10690130 TI - Haemodynamic effects of diaspirin crosslinked haemoglobin (DCLHb) given before abdominal aortic aneurysm surgery. AB - We studied 34 patients undergoing elective repair of an abdominal aortic aneurysm under combined general anaesthesia and epidural block to evaluate the acute effects of diaspirin crosslinked haemoglobin (DCLHb) 50, 100 and 200 mg kg-1 i.v. Haemodynamic variables were measured continuously using pulmonary and radial artery catheters, and oxygen delivery and consumption were calculated at regular intervals. DCLHb was shown to be vasoactive, producing an increase in mean arterial pressure of approximately 25% with each dose, with small decreases in cardiac index and calculated oxygen delivery. These effects persisted beyond the end of infusion and provided a degree of cardiovascular stability during the operative procedure. The effects of DCLHb on oxygen consumption at these doses were minimal. PMID- 10690131 TI - High frequency jet ventilation and gas trapping. AB - We have compared three types of high frequency jet ventilation (HFJV) with conventional positive pressure ventilation in patients recovering from elective coronary artery bypass surgery. Twelve patients were allocated randomly to receive HFJV at ventilatory frequencies of 60, 100, 150 and 200 bpm from a standard jet ventilator at either the proximal or distal airway (HFJV.p and HFJV.d), or from a valveless high frequency jet ventilator acting as a pneumatic piston (VPP). Trapped gas volume (Vtr), cardiac index (CI) and right ventricular ejection fraction (RVEF) were measured. Vtr was related to the type of HFJV used (P < 0.05) and ventilatory frequency (P < 0.05). CI decreased with increasing rate of HFJV (P < 0.05) and there were significant differences between the three types of HFJV (P < 0.05). RVEF showed a linear relationship with ventilatory frequency (P < 0.05) decreasing most with the VPP. The decrease in RVEF was associated with an increase in right ventricular end-systolic volume (P < 0.05) suggesting that an increase in right ventricular afterload was the cause. The same three types of HFJV were compared using a lung model with variable values of compliance and resistance, to assess the impact of lung mechanics on gas trapping (Vtr, ml). Lung model compliance (C) was set at 50 or 25 ml cm H2O-1 and resistance (R) at 5 or 20 cm H2O litre-1 s, where values of 50 and 5, respectively, are normal. Vtr increased with ventilatory frequency for all types of jet ventilation (P < 0.05), varying with the type of jet ventilation used (P < 0.05). PMID- 10690132 TI - Effect of omitting regular ACE inhibitor medication before cardiac surgery on haemodynamic variables and vasoactive drug requirements. AB - Adverse events during coronary artery bypass graft (CABG) surgery have been described in patients receiving angiotensin converting enzyme (ACE) inhibitors, including hypotension on induction of anaesthesia and an increase in vasoconstrictor requirements after cardiopulmonary bypass (CPB). Omitting regular ACE inhibitor medication before surgery may improve cardiovascular stability during anaesthesia. We evaluated prospectively the effect of omitting regular ACE inhibitor medication before CABG surgery on haemodynamic variables and use of vasoactive drugs. We studied 40 patients with good left ventricular function, allocated randomly to omit or continue ACE inhibitor medication before surgery. Arterial pressure, cardiac output, systemic vascular resistance and use of vasoactive drugs were recorded during anaesthesia and in the early postoperative period. Patients who omitted their ACE inhibitors had greater mean arterial pressure during the study and required less vasopressors during CPB. However, these patients required more vasodilators to control hypertension after CPB and in the early postoperative period. There was no difference in hypotension on induction of anaesthesia or in the use of vasoconstrictors after CPB. We conclude that omitting ACE inhibitors before surgery did not have sufficient advantage to be recommended routinely. PMID- 10690133 TI - Haemodynamic effects of rapacuronium in adults with coronary artery or valvular disease. AB - We have assessed the haemodynamic effects of rapacuronium (Org 9487) in adults undergoing cardiac surgery and compared these with vecuronium and placebo. We studied 56 adult patients undergoing coronary artery bypass grafting or valve replacement surgery using a fentanyl-based anaesthetic technique. A pulmonary artery flotation catheter was inserted before induction of anaesthesia. After induction, tracheal intubation and stabilization of haemodynamic measurements, subjects were allocated randomly to receive rapacuronium 1.5 mg kg-1 vecuronium 0.1 mg kg-1 or saline placebo. Haemodynamic measurements were made before drug administration and 1, 3, 5 and 10, and if possible, 15 min after administration. Rapacuronium was associated with statistically significant increases in heart rate (17%) and cardiac index (15%) and decreases in mean arterial pressure (11%) and systemic vascular resistance (18%), whereas vecuronium and placebo were associated with significant decreases in heart rate only (14-15%) (P < 0.05). No cutaneous signs of histamine release were observed. Clinically, the results were within acceptable limits. Our results suggest that administration of rapacuronium may be associated with significant changes in heart rate and arterial pressure in patients undergoing coronary artery bypass grafting. PMID- 10690134 TI - Antagonism of rapacuronium using edrophonium or neostigmine: pharmacodynamics and pharmacokinetics. AB - We have studied the pharmacodynamics and pharmacokinetics of rapacuronium (Org 9487) in 70 healthy patients. Neuromuscular transmission was monitored using TOF stimulation of the ulnar nerve and mechanomyography of the adductor pollicis muscle. Half of the patients were given a single dose of rapacuronium 1.5 mg kg-1 and the remainder rapacuronium 1.5 mg kg-1 with three incremental doses of 0.5 mg kg-1, each given when T1/T0 had recovered to 25%. In all patients, neuromuscular block was antagonized using neostigmine 0.05 mg kg-1 or edrophonium 1.0 mg kg-1 (allocated randomly), 2 min after the final dose of rapacuronium. All patients developed complete block after rapacuronium 1.5 mg kg-1. Mean onset time was 66 (SD 24) s. In patients who received an antagonist 2 min after the first dose of rapacuronium, time to recovery of T1/T0 to 25% was similar after neostigmine (9.8 (3.8) min) and edrophonium (10.3 (4.3) min): in patients who received incremental doses of rapacuronium, spontaneous recovery of T1/T0 to 25% after the first dose was 18.9 (4.7) min. In those who received an antagonist 2 min after the first dose of rapacuronium, times to recovery of T4/T1 to 0.7 were also similar after neostigmine (23.7 (7.7) min) and edrophonium (29.1 (10.7) min). After three incremental doses of rapacuronium, there was a longer time to recovery of T1/T0 = 25% after neostigmine compared with edrophonium (5.1 (1.0) vs 3.3 (1.3) min; P < 0.05) but more rapid recovery to T1/T0 = 75% (10.1 (2.9) vs 16.8 (10.1) min; P < 0.05) and T4/T1 = 0.7 (19.8 (6.3) vs 35.1 (10.4) min; P < 0.05). A three compartment pharmacokinetic model was justified. Typical values for clearance and initial volume of distribution (V1) were 4.4 ml kg-1 min-1 and 94.8 ml kg-1, respectively. In females, clearance was decreased by 38.5% compared with males and V1 was decreased by 25% in patients aged more than 65 yr. PMID- 10690135 TI - Sevoflurane anaesthesia causes a transient decrease in aquaporin-2 and impairment of urine concentration. AB - Sevoflurane anaesthesia is occasionally associated with polyuria, but the exact mechanism of this phenomenon has not been clarified. Aquaporin-2 (AQP2) is an arginine vasopressin (AVP)-regulated water channel protein localized to the apical region of renal collecting duct cells and is involved in the regulation of water permeability. To elucidate the effect of sevoflurane anaesthesia on urine concentration and AQP2, we have compared serum and urinary concentrations of AVP, AQP2 and osmolar changes during sevoflurane and propofol anaesthesia. General anaesthesia was induced with sevoflurane or propofol in 30 patients for a variety of major surgical procedures. Blood and urine samples were obtained from patients at baseline, and 90 and 180 min after induction of anaesthesia. AVP and AQP2 concentrations were measured by radioimmunoassay. In both groups, plasma and urinary concentrations of AVP increased similarly during anaesthesia although plasma osmolality remained unchanged. Although urinary AQP2 excretion in the propofol group increased together with changes in plasma and urinary AVP, urinary AQP2 was significantly lower at 90 min in the sevoflurane group. Urine osmolality in the sevoflurane group also showed a transient but significant decrease in parallel with suppression of AQP2. Our data suggest that sevoflurane anaesthesia transiently produced an impaired AQP2 response to an increase in intrinsic AVP. PMID- 10690136 TI - The Haldane effect--an alternative explanation for increasing gastric mucosal PCO2 gradients? AB - When venous oxygen saturation increases as a result of increased blood flow, changes in venous blood PCO2 and carbon dioxide content may differ because of the Haldane effect. The Haldane effect may also explain increases in gastric mucosal arterial PCO2 gradient despite major increases in splanchnic blood flow. We re analysed data from 22 patients after cardiac surgery who were randomized to receive either dobutamine or placebo, and a separate group of patients who received dobutamine for low cardiac output (n = 6). Three different values of gastric mucosal oxygen extraction at baseline were assumed (0.3, 0.5 and 0.7). In nine of 14 patients with both increasing splanchnic blood flow and mucosal arterial PCO2 gradient, an equal increase in mucosal and total splanchnic blood flow, oxygen consumption and carbon dioxide production together with the Haldane effect would have caused an increase in mucosal-arterial PCO2 gradients from a mean value of 0.53 (SD 0.88) kPa at baseline to 0.68-0.82 (0.89-0.90) kPa (P < 0.01). In the remaining patients, disproportionate changes in flow and metabolism must have been involved in addition to the Haldane effect. We conclude that whenever major changes in mucosal tissue oxygen extraction are likely to occur, an increase in the mucosal-arterial PCO2 gradient may be explained in part or completely by the Haldane effect, and may therefore not reflect worsening perfusion. PMID- 10690137 TI - Continuous auditory monitoring--how much information do we register? AB - We have studied response times of 30 anaesthetists to a standardized episode of arterial oxygen desaturation in a simulated patient, randomized to the use of either a fixed or variable pitch pulse oximeter. We wished to determine if a variable auditory signal was important in detecting adverse events. A variable pitch pulse signal had a shorter time to recognition of desaturation (P < 0.0001), with a mean response time of 32 s, compared with 129 s for the fixed pitch signal. PMID- 10690138 TI - Comparison of 0.25% S(-)-bupivacaine with 0.25% RS-bupivacaine for epidural analgesia in labour. AB - We have compared the efficacy of 0.25% S(-)-bupivacaine with 0.25% RS-bupivacaine in providing epidural analgesia for labour in a randomized, multicentre, double blind study. Analgesia was initiated with 10 ml of the study solution and maintained with 10-ml top-ups. We studied 137 women and treatments were found to be equivalent for onset, duration and quality of block. Median onset of pain relief was 12 min for both drugs and median duration was 49 (range 3-129) min and 51 (7-157) min for S(-)-bupivacaine and RS bupivacaine, respectively. The estimated treatment difference for duration of pain relief was -4 (90% CI -13, 6) min. Thirty patients failed to achieve pain relief after the first injection (20 patients after S(-)-bupivacaine and 10 after RS-bupivacaine; P = 0.039). However, median duration of pain relief from the first top-up was 82 (range 3-164) min for S(-)-bupivacaine and 76 (22-221) min for RS-bupivacaine. There were no significant differences in the quality of analgesia, as assessed by the investigators. There were no significant differences in the extent of sensory block, percentage of patients with motor block or incidence of adverse events. PMID- 10690139 TI - Comparison of low-dose epidural with combined spinal-epidural analgesia for labour. AB - We have performed a randomized comparison of two low-dose epidural regimens for analgesia in labour, differing only in the manner in which initial analgesia was established. In the epidural (EPI) group, 484 women received a loading dose of 20 ml of 0.1% bupivacaine with fentanyl 2 micrograms ml-1. In the combined spinal epidural (CSE) group, 524 women received a spinal injection of plain bupivacaine 2.5 mg with fentanyl 25 micrograms. In both groups, these initial doses were followed by 0.1% bupivacaine with fentanyl 2 micrograms ml-1 infused at a rate of 12 ml h-1, with 20-ml top-ups for breakthrough pain. The groups were compared for midwife assessment of analgesic efficacy, delivery mode, patient assessments of first stage analgesia, second stage analgesia, overall analgesia, motor block and complications. Midwives, who were not blinded to the treatment groups, assessed 61.6% of CSE as providing 'excellent' analgesia compared with 56.4% of epidurals (P = 0.02). Patients assessed overall analgesia as 'excellent' in 74.8% of CSE compared with 71.7% of epidurals (P = 0.14). Other comparisons between groups revealed no differences. These findings may have been affected by an uneven distribution of multiparous women between the groups (25% in the EPI group and 34.2% in the CSE group; P = 0.002). However, subgroup analysis of primiparous and multiparous women did not alter the results. PMID- 10690140 TI - Metoclopramide in the prevention of postoperative nausea and vomiting: a quantitative systematic review of randomized, placebo-controlled studies. AB - Metoclopramide has been used for almost 40 yr to prevent postoperative nausea and vomiting (PONV). We have reviewed the efficacy and safety of metoclopramide for the prevention of PONV. A systematic search (MEDLINE, EMBASE, manufacturers' databases, hand searching, bibliographies, all languages, up to June 1998) was performed for full reports of randomized comparisons of metoclopramide with placebo in surgical patients. Relevant end-points were prevention of early PONV (within 6 h after operation), late PONV (48 h) and adverse effects. Combined data were analysed using relative benefit/risk and number-needed-to-treat/harm. In 66 studies, 3260 patients received 18 different regimens of metoclopramide, and 3006 controls received placebo or no treatment. There was no evidence of dose responsiveness with oral, i.m., intranasal or i.v. metoclopramide in children and adults. In adults, the best documented regimen was 10 mg i.v. There was no significant anti-nausea effect. The numbers-needed-to-treat to prevent early and late vomiting were 9.1 (95% confidence intervals 5.5-27) and 10 (6-41), respectively. In children, the best documented regimen was 0.25 mg kg-1 i.v. The number-needed-to-treat to prevent early vomiting was 5.8 (3.9-11). There was no significant late anti-vomiting effect. Minor drug-related adverse effects (sedation, dizziness, drowsiness) were not significantly associated with metoclopramide. There was one adult who experienced extrapyramidal symptoms with metoclopramide. PMID- 10690141 TI - Dexamethasone reduces nausea and vomiting after laparoscopic cholecystectomy. AB - We have evaluated the antiemetic effect of i.v. dexamethasone compared with saline in the prevention of nausea and vomiting after laparoscopic cholecystectomy. We studied 90 patients requiring general anaesthesia for laparoscopic cholecystectomy, in a randomized, double-blind, placebo-controlled study. The dexamethasone group (n = 45) received dexamethasone 8 mg i.v. and the saline group received saline 2 ml i.v. at induction of anaesthesia. Anaesthesia was maintained with isoflurane in oxygen. We found that 10% of patients in the dexamethasone group compared with 34% in the saline group reported vomiting (P < 0.05). Of note, the total incidence of nausea and vomiting was 23% in the dexamethasone group and 63% in the saline group (P < 0.001). We conclude that dexamethasone 8 mg significantly decreased the incidence of nausea and vomiting after laparoscopic cholecystectomy. PMID- 10690142 TI - Hospital admission after day-case gynaecological laparoscopy. AB - We have examined aspects of the anaesthetic technique that may influence the likelihood of unplanned overnight hospital admission after ambulatory gynaecological laparoscopy and have determined if any anaesthetically controllable factors were involved. The retrospective audit involved 300 patients. All patients attended the day-case unit at the Liverpool Women's Hospital between September 1996 and May 1997. One hundred ASA I-II patients who had unplanned overnight admissions during this time were evaluated. For every admitted patient, two similar patients who did not require admission were studied. Variables such as patient age and anaesthetic technique were evaluated by logistic regression. Our results indicated that postoperative emesis was the commonest cause for admission. Significant factors increasing the likelihood of unplanned admission included returning from the recovery unit after 15:00, use of a laryngeal mask airway and undergoing diagnostic laparoscopy. Significant factors reducing the likelihood of admission were the use of fentanyl and rectally administered diclofenac. PMID- 10690143 TI - Effects of tramadol on minimum alveolar concentration (MAC) of isoflurane in rats. AB - It has been suggested previously that tramadol increases central nervous system activity and 'lightens' anaesthesia with volatile agents. We assessed the effects of tramadol on the minimum alveolar concentration (MAC) of isoflurane in 56 Wistar rats, instrumented chronically with an arterial and central venous catheter. The MAC of isoflurane was determined using the tail clamp method under three conditions: (1) after injection of saline (control); (2) after administration of tramadol 10 mg kg-1 i.v.; and (3) after administration of morphine 1 mg kg-1 i.v. The studies were repeated after treatment with the antagonists naloxone or yohimbine. Tramadol and morphine both reduced the MAC of isoflurane from mean 1.38 (SEM 0.05)% to 1.22 (0.06)% and 1.17 (0.06)%, respectively (P < 0.05). Concomitant administration of yohimbine did not abolish this reduction in MAC. In contrast, after pretreatment with naloxone, tramadol (1.47 (0.04)%) or morphine (1.38 (0.07)%) did not cause a reduction in the MAC of isoflurane compared with controls (1.39 (0.06)%). We conclude that tramadol and morphine reduced the MAC of isoflurane to a small but significant extent. For both drugs, this effect was related to their action at opioid receptors. PMID- 10690144 TI - Catecholaminergic activity and 3',5'-cyclic adenosine monophosphate concentrations in the right ventricle after acute and chronic morphine administration in the rat. AB - We have examined possible regulation of norepinephrine and dopamine concentrations and turnover in the right ventricle of the rat after acute administration of saline i.p. or morphine 30 mg kg-1 i.p. to placebo (naive) or morphine (tolerant) pretreated rats. We also assessed concentrations of 3',5' cyclic adenosine monophosphate (cAMP) in the right ventricle after the same treatments. Concentrations of catecholamines and their metabolites in the heart were measured by high-pressure liquid chromatography with electrochemical detection (HPLC/DE). Concentrations of cAMP in the heart were measured by radioimmunoassay (RIA). Administration of morphine to naive rats did not modify concentrations of norepinephrine (NE), normetanephrine (NMN) or NMN/NE ratio in the right ventricle. However, dopamine concentrations increased whereas dopamine turnover decreased. In addition, cAMP concentrations decreased after acute administration morphine to naive rats. In rats pretreated with morphine chronically, there was an increase in norepinephrine concentrations with no change in normetanephrine concentrations or norepinephrine turnover after acute injection of morphine. In contrast, dopamine turnover increased in the tolerant groups after acute injection of saline or morphine compared with the nave group given morphine, indicating that tolerance develops to the acute effects of the opioid. Concentrations of cAMP increased after chronic morphine administration. Our results demonstrate that chronic morphine pretreatment leads to up-regulation of the cAMP system in the heart and suggest that this up-regulation may be involved in the cellular mechanisms implicated in the adaptive changes of dopaminergic neurones in the heart observed during chronic treatment with morphine. PMID- 10690145 TI - Ultrasound: an emerging role in anaesthesia and intensive care. PMID- 10690146 TI - Oxygen administration and explicit memory: no improvement found in healthy volunteers. AB - It has been suggested that oxygen administration to healthy volunteers could improve their memory. We tested this hypothesis with a twin, double crossover, placebo-controlled study in 20 healthy non-smokers, allocated randomly to one of two groups. Blinded to the nature of the gas, group A breathed air first then oxygen on day 1, and then oxygen first, followed by air on day 2. Group B had all exposures in reverse order. After each gas exposure a written memory test with a list of 20 words was carried out and evaluated by a blinded observer. Recall after oxygen exposure (mean 8.3 words) was not significantly different from that after air exposure (mean 9 words). PMID- 10690147 TI - Evaluation of pressure changes in a new design tracheal tube cuff, the Portex Soft Seal, during nitrous oxide anaesthesia. AB - We have measured pressure changes in a newly designed tracheal tube cuff, the Portex Soft Seal, during nitrous oxide anaesthesia compared with a Mallinckrodt Lo-Contour tube and a Portex Profile tube. The pressure increases in both control groups were significantly greater than those with the new design (P < 0.0001 in each case). The mean increase in pressure in the Mallinckrodt Lo-Contour tube cuff was 9.9 (SD 3.4) mm Hg compared with 10.3 (1.8) mm Hg in the Portex Profile tube cuff and 2.1 (1.5) mm Hg in the Portex Soft Seal tube cuff. We conclude that the Portex Soft Seal cuff prevented a significant increase in intracuff pressure during nitrous oxide anaesthesia. PMID- 10690148 TI - Posterior epidural space depth: safety of the loss of resistance and hanging drop techniques. AB - We have compared skin to epidural space distance (SED) and tip to tip distance (TTD), a measure of posterior epidural space depth (PESD), in 40 patients with a 27-gauge Whitacre needle after identification of the epidural space using the hanging drop (HD) or loss of resistance (LOR) to air technique. After the LOR technique, TTD was found to be 2 mm greater than that after the HD technique, whereas SED was the same. We conclude that identification of the epidural space can be performed successfully with both techniques, but with a diminished risk of dural damage after LOR compared with the HD technique. PMID- 10690149 TI - Self-prepared heparinized syringes for measuring ionized magnesium in critical care patients. AB - We have compared ionized magnesium assays in the Nova 8 electrolyte analyser using dry balanced heparinized syringes and self-prepared heparinized syringes. Thirty blood specimens were obtained into syringes either operator-prepared with liquid sodium heparin or commercially manufactured dry balanced heparinized syringes. There was a good correlation between results from the two syringes. The mean difference between sampling methods was 0.01 mmol litre-1 (95% confidence index -0.05 to 0.08 mmol litre-1). The correlations for sodium, potassium and ionized calcium assays were similarly close. The relationship between sampling methods was close enough to justify the clinical use of self-prepared syringes, with potential economies in clinical costs. PMID- 10690150 TI - Ondansetron and droperidol in the prevention of postoperative nausea and vomiting. AB - We have performed a prospective, randomized, double-blind clinical study to assess the efficacy of ondansetron, droperidol, or both, in preventing postoperative emesis. We studied 242 patients undergoing biliary or gynaecological surgery under general anaesthesia. Shortly before induction of anaesthesia, patients received: saline i.v. (group I, n = 62); droperidol 2.5 mg i.v. (group 2, n = 60); ondansetron 4 mg i.v. (group 3, n = 57); or droperidol 2.5 mg with ondansetron 4 mg i.v. (group 4, n = 63). Nausea occurred in 45%, 37%, 32% and 29% (P = 0.234) and vomiting in 23%, 17%, 9% and 5% (P = 0.016) of patients in groups 1, 2, 3 and 4, respectively, during the first 24 h. Groups 2 and 4 had greater sedation scores than group 1 during the first 3 h (P < 0.01). We conclude that both droperidol and ondansetron showed a significant antiemetic effect, ondansetron was not significantly better than droperidol, and the combination of droperidol and ondansetron was better than droperidol but no better than ondansetron alone. PMID- 10690151 TI - Development of rapid atrial fibrillation with a wide QRS complex after neostigmine in a patient with intermittent Wolff-Parkinson-White syndrome. AB - We report the case of a 67-yr-old man with intermittent Wolff-Parkinson-White (WPW) syndrome in whom neostigmine produced life-threatening tachyarrhythmias. The patient was scheduled for microsurgery for a laryngeal tumour. When he arrived in the operating room, the electrocardiogram showed normal sinus rhythm with a rate of 82 beat min-1 and a narrow QRS complex which remained normal throughout the operative period. On emergence from anaesthesia, the sinus rhythm (87 beat min-1) changed to atrial fibrillation with a rate of 80-120 beat min-1 and a normal QRS complex. We did not treat the atrial fibrillation because the patient was haemodynamically stable. Neostigmine 1 mg without atropine was then administered to antagonize residual neuromuscular block produced by vecuronium. Two minutes later, the narrow QRS complexes changed to a wide QRS complex tachycardia with a rate of 110-180 beat min-1, which was diagnosed as rapid atrial fibrillation. As the patient was hypotensive, two synchronized DC cardioversions of 100 J and 200 J were given, which restored sinus rhythm. No electrophysiological studies of anticholinesterase drugs have been performed in patients with WPW syndrome. We discuss the use of these drugs in this condition. PMID- 10690152 TI - Paravertebral block in the management of liver capsule pain after blunt trauma. AB - We present a case of liver capsule pain after blunt abdominal trauma. The patient was unable to tolerate patient-controlled i.v. opioids, and epidural infusion of local anaesthetic was considered undesirable because of the potential risk of complications. Pain was managed successfully with paravertebral infusion of local anaesthetic at the right T10 level. Innervation of the liver and possible mechanisms of visceral pain processing are discussed. PMID- 10690153 TI - Opioid-induced pruritus: repeated vs single dose ondansetron administration in preventing pruritus after intrathecal morphine. PMID- 10690154 TI - Effect of rocuronium compared with succinylcholine on IOP. PMID- 10690155 TI - Effect of rocuronium compared with succinylcholine on IOP. PMID- 10690156 TI - Intraoperative therapeutic suggestions. PMID- 10690157 TI - Glucose utilization dynamics and food intake. PMID- 10690158 TI - Wheat bran supplementation does not affect biochemical markers of bone turnover in young adult women with recommended calcium intake. AB - We investigated the effect of wheat bran on biochemical indicators of Ca and bone metabolism in nineteen healthy women, aged 25.5 (SE 0.9) years. Subjects received six wheat bran biscuits or six white flour biscuits per day for a period of 4 weeks (crossover). Wheat bran consumption increased fibre intake from 17.7 (SE 1.3) to 29.6 (SE 1.3) g/d (7 d food record) and enhanced P intake from 1225 (SE 59) mg/d to 1663 (SE 65) mg/d; P < 0.001. Mean daily Ca intake during wheat bran consumption (1110 (SE 82) mg/d) significantly (P = 0.008) exceeded Ca ingestion during the white flour period (955 (SE 67) mg/d). Wheat bran increased the number of defecations per week from 7.9 (SE 0.8) to 12.2 (SE 1.4) (P = 0.0018). Urinary Ca excretion over 24 h significantly (P = 0.021) decreased from 473 (SE 53) mumol/mmol creatinine (control period) to 339 (SE 37) mumol/mmol creatinine (wheat bran period). Serum 25-hydroxyvitamin D, 2 h fasting urinary Ca/creatinine excretions and 24 h urinary P excretion remained constant. No differences in serum levels of carboxy-terminal propeptide of type I procollagen (biomarker of bone formation) or in 2 h fasting urinary hydroxyproline/creatinine excretions (biomarker of bone resorption) were observed at the end of the two cycles of dietary supplementation. We conclude that a high fibre intake of approximately 30 g/d has no significant adverse effects on bone turnover in subjects with Ca intakes above 1000 mg/d and that the reduction in 24 h urinary Ca excretion is most probably the result of an adaptation process, induced by a decrease in net absorbed Ca. PMID- 10690159 TI - Blood glucose and meal patterns in time-blinded males, after aspartame, carbohydrate, and fat consumption, in relation to sweetness perception. AB - In a study of the impact of aspartame, fat, and carbohydrate on appetite, we monitored blood glucose continuously for 431 (SE 16) min. Ten healthy males (19 31 years) participated in three time-blinded visits. As blood glucose was monitored, appetite ratings were scored at randomized times. On the first meal initiation, volunteers consumed one of three isovolumetric drinks (aspartame, 1 MJ simple carbohydrate, and 1 MJ high-fat; randomized order). High-fat and high carbohydrate foods were available ad libitum subsequently. Blood glucose patterns following the carbohydrate drink (+1.78 (SE 0.28) mmol/l in 38 (SE 3) min) and high-fat drink (+0.83 (SE 0.28) mmol/l in 49 (SE 6) min) were predictive of the next intermeal interval (R 0.64 and R 0.97 respectively). Aspartame ingestion was followed by blood glucose declines (40% of subjects), increases (20%), or stability (40%). These patterns were related to the volunteers' perception of sweetness of the drink (R 0.81, P = 0.014), and were predictive of subsequent intakes (R -0.71, P = 0.048). For all drinks combined, declines in blood glucose and meal initiation were significantly associated (chi 2 16.8, P < 0.001), the duration of blood glucose responses and intermeal intervals correlated significantly (R 0.715, P = 0.0001), and sweetness perception correlated negatively with hunger suppression (R -0.471, P = 0.015). Effects of fat, carbohydrate, and aspartame on meal initiation, meal size, and intermeal interval relate to blood glucose patterns. Varied blood glucose responses after aspartame support the controversy over its effects, and may relate to sweetness perception. PMID- 10690160 TI - The effects of sucrose and maize oil on subsequent food intake and mood. AB - The effects of sucrose and oil preloads were explicitly compared in a single blind controlled trial using a between-subjects design. Eighty adult subjects (forty-three male, thirty-seven female) aged 18-50 years received at 11.00 hours one of four yoghurt preloads. All were 80 g low-fat, unsweetened yoghurt (188 kJ), containing additionally (1) saccharin (control, 23 kJ), or (2) 40 g sucrose (859 kJ), (3) 40 g maize oil (1569 kJ), (4) 20 g sucrose, 20 g maize oil (1213 kJ). Subjects were normal eaters and of normal weight (male mean weight: 68.8 (SD 3.2) kg, BMI 21.8 (SD 1.6) kg/m2; female mean weight: 53.7 (SD 5.1) kg, BMI 20.4 (SD 1.2) kg/m2). Food intake was measured with a food diary and mood with ten single Likert scales. ANOVA was conducted using preload type (saccharin, sucrose, oil, sucrose + oil), sex (male, female) and early v. late breakfast times as factors. Mood was analysed using the same design, with time of rating (immediate, 60 min, 120 min) as an additional factor. Men ate more after the saccharin preload than after the other preloads, but did not vary the time of their next solid food. Women increased the intermeal interval only after the oil preload, which also had the highest energy content value, but did not vary the energy content of their next solid food. The saccharin preload decreased rated tiredness at 2 h compared with the sucrose preload, possibly due to its lower energy content. The preloads containing sucrose or sucrose + oil increased calmness between 1 and 2 h afterwards, compared with the saccharin preload. It is concluded that both sucrose and oil increase the intermeal interval in men, but that women are less sensitive to preloading. The mood effects suggest that tiredness after carbohydrate at 2 h may in part be a decrease in rated energy compared with the increased rated energy found after a preload with low energy content. Carbohydrate may genuinely increase calmness. These effects apply to non restrained eaters of normal weight. PMID- 10690161 TI - Carbohydrate intake improves cognitive performance of stress-prone individuals under controllable laboratory stress. AB - Cognitive performance has been found to decline after exposure to stress, particularly in stress-prone subjects. The present study investigated whether a carbohydrate-rich, protein-poor (CR/PP) diet, which may enhance cerebral serotonin function in stress-prone subjects due to increases in the available tryptophan, improves the performance of stress-prone subjects after exposure to acute laboratory stress. Twenty-two high-stress-prone (HS) subjects and twenty one low-stress-prone (LS) subjects aged between 19 and 26 years performed a memory scanning task after controllable and uncontrollable stress, following either a CR/PP diet or a protein-rich, carbohydrate-poor (PR/CP) isoenergetic diet. Uncontrollable stress reduced feelings of control (F(1,38) 9.30; P = 0.004), whereas pulse rate and skin conductance increased after both stress tasks (F(1,38) 78.34; P = 0.0005 and F(1,37) 83.16; P = 0.0004). Diet, stress-proneness and stress-controllability interacted (F(1,36) 9.46; P = 0.004) in such a way that performance in HS subjects was better with the CR/PP diet than with the PR/CP diet, but only after controllable stress. As the CR/PP diet has been found to increase the plasma tryptophan:large neutral amino acids ratio, indicating an increased availability of cerebral tryptophan and, thus, higher serotonin levels, it appears that there may be an increased availability of brain serotonin in HS subjects after controllable laboratory stress. PMID- 10690162 TI - The effect of palm oil, lard, and puff-pastry margarine on postprandial lipid and hormone responses in normal-weight and obese young women. AB - Only a few studies have been published on the postprandial effects of different fatty acids in obese subjects. Therefore, the present study investigated the effects of three test meals containing palm oil (PO), lard (LD), or puff-pastry margarine (PPM), all normal dietary ingredients, on postprandial lipid and hormone responses in normal-weight and obese young women. The study was performed as a randomized, crossover design. The fats differed in the content of palmitic acid, stearic acid, and trans monounsaturated fatty acids allowing a dietary comparison of different 'solid' fatty acids. The obese women had significantly higher fasting concentrations and postprandial responses of plasma total triacylglycerol (TAG), chylomicron-TAG, and insulin compared with the normal weight women but there was no significant difference in the postprandial responses between the three test meals. The obese women had fasting concentrations of leptin four times greater than the normal-weight women. There were no postprandial changes in the concentrations of leptin. The fasting concentrations of HDL-cholesterol were significantly lower in the obese women than in the normal-weight women, whereas there was no significant difference between the two groups in the concentrations of total cholesterol or LDL cholesterol. These results provide evidence that obese women have exaggerated lipid and hormone responses compared with normal-weight women but the different contents of saturated and trans monounsaturated fatty acids provided by PO, LD, and PPM have no effect in either group. PMID- 10690163 TI - Incorporation and washout of orally administered n-3 fatty acid ethyl esters in different plasma lipid fractions. AB - The aim of the present study was to quantify the incorporation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) into plasma lipids after oral administration of n-3 fatty acid ethyl esters, since little is known about the rate and pattern of incorporation into plasma lipid fractions. In addition, we aimed to obtain preliminary information regarding EPA half-life, which is needed to establish an optimal dosing schedule. Five healthy volunteers ingested two 8.5 g doses of n-3 fatty acid ethyl esters daily for 7 d, supplying 6.0 g EPA/d and 5.3 g DHA/d. The fatty acid compositions of plasma phospholipids (PL), cholesteryl esters (CE) and triacylglycerols (TAG) were determined during supplementation and during a washout period of 7 d. Half-lives of EPA and DHA were calculated. The proportion of EPA in PL showed a 15-fold increase after 7 d (P < 0.001), while DHA showed a smaller increase (P < 0.01). In CE, EPA also increased (P < 0.05), while DHA did not increase at all. Remarkably, incorporation of DHA into TAG was even higher than that of EPA. Half-life of EPA in PL ranged from 1.63 to 2.31 d (mean 1.97 (SE 0.15) d), whereas mean half-life of EPA in CE was 3.27 (SE 0.56) d. In three subjects, washout of EPA and DHA from TAG seemed to follow a bi-exponential pattern, with a short half-life (< 1 d) in the initial phase and a half-life of several days in the second phase. In conclusion, EPA ethyl esters are rapidly incorporated into plasma lipids, especially into PL. The relatively long half-life of EPA in plasma would permit a dosing schedule with intervals of > or = 12 h in supplementation studies. PMID- 10690164 TI - The effect of a model melanoidin mixture on faecal bacterial populations in vitro. AB - The Maillard reaction produces coloured, macromolecular materials (melanoidins) in a variety of foods, on heating. Significant quantities may enter the human gut on a daily basis, but there is little information on their metabolism in the human colon. As the large bowel contains a diverse population of bacteria involved in normal bowel function, it is possible that melanoidins are metabolized therein. Depending on the bacteria involved, there may be disease or health implications. The aim of the present study was to use in vitro models to determine the digestibility of melanoidins and the effect of melanoidins on colonic bacteria in the gastrointestinal tract. Melanoidins were prepared and the effects of simulated upper-gut secretions on their stability determined in a model system. The effects of faecal bacteria were also determined, in batch culture, with a combination of phenotypic and genotypic (probes) criteria being used to identify the microbial diversity involved. Simulation of peptic and pancreatic digestion showed that the melanoidins did not produce detectable amounts of low-molecular-mass degradation products. However, melanoidins affected the growth of gut bacteria during mixed culture growth. The effect was to cause a non-specific increase in the anaerobic bacteria enumerated. This in vitro study indicates that melanoidins can affect the growth of human large-bowel bacteria and serves to demonstrate possible effects that may occur in vivo. Given the large and varied number of food items that contain Maillard reaction products, this may have relevance for lower-gut health. PMID- 10690165 TI - Involvement of lipoic acid in plasma metabolites, hepatic oxygen consumption, and metabolic response to a beta-agonist in broiler chickens. AB - The present study was conducted to determine the role of alpha-lipoic acid (LA) in plasma metabolites, hepatic O2 consumption, and beta-adrenergic response in broilers. In Expt 1, 12-d-old female broiler chicks were divided into three dietary groups and fed on diets with or without LA (5 or 50 mg/kg) until 4 or 6 weeks of age, as a 2 x 3 factorial arrangement. The dietary LA had no effect on growth rates (body weight, abdominal fat, breast muscle, and liver). The higher level of LA increased plasma non-esterified fatty acid and decreased plasma triacylglycerol concentrations only at 6 weeks of age. A 42% increase in hepatic respiration was observed in the 4-week-old chickens given 50 mg LA/kg diet. In Expt 2, 3-d-old female broiler chicks were treated with or without dietary LA at 50 mg/kg. At 30 and 31 d old, isoproterenol (2 mg/kg body weight per h) was continuously infused into a wing vein for 2 h, and changes in plasma glucose, triacylglycerol, and non-esterified fatty acid concentrations were analysed. Isoproterenol increased plasma glucose over basal levels maximally at 60 min. Furthermore, the glucose increase in the LA-treated chickens was 35% greater than that of the controls at this time. Plasma non-esterified fatty acid and triacylglycerol concentrations were decreased by the isoproterenol infusion, regardless of LA administration. Therefore, the present study suggests that dietary LA has repartitioning effects on energy metabolism in chickens (although this depends on age-related metabolic state) and is a possible facilitator in the beta-adrenergic response of plasma glucose to a beta-agonist. PMID- 10690166 TI - Prostate cancer prevention: review of target populations, pathological biomarkers, and chemopreventive agents. PMID- 10690167 TI - The neuropathology of the vegetative state after head injury. PMID- 10690168 TI - Clinical application of the Quantiplex HCV RNA 2.0 and Amplicor HCV Monitor assays for quantifying serum hepatitis C virus RNA. AB - AIM: To compare the performance characteristics and clinical application of two different technologies for quantifying serum hepatitis C virus (HCV) RNA levels. METHODS: HCV RNA was quantified by Amplicor HCV Monitor assay (Amplicor) and Quantiplex HCV RNA 2.0 assay (bDNA-2) in 119 sera from 107 HCV infected patients. RESULTS: Both assays had similar sensitivity (79.4% for Amplicor; 86.0% for bDNA 2), acceptable coefficients of variation (5.3% in Amplicor; 2.6% in bDNA-2), and good linearity (r2 > or = 0.98). There was a positive correlation between quantification values of both methods (r = 0.683, p < 0.001). The Amplicor values were on an average 1.76 log lower than bDNA-2 results. Male subjects and HCV genotype 1b were significantly associated with higher viral load determined by Amplicor, but not with viral load measured by bDNA-2. In 70 chronic HCV infected patients treated with interferon alfa, mean (SD) pretreatment viral load in 27 complete responders (3.47 (0.84) logs for Amplicor, 5.63 (0.58) for bDNA-2) was significantly lower than in non-responders (4.43 (1.01) logs for Amplicor, 6.10 (0.67) logs for bDNA-2; p < 0.001). Cut off points of 3.9 logs for Amplicor and 5.8 logs for bDNA-2 were determined to be the best for predicting response to interferon alfa, giving acceptable sensitivity (70.4%, 74.1%), specificity (72.1%, 65.1%), and accuracy (71.4%, 68.6%), respectively. CONCLUSIONS: Both the Amplicor and bDNA-2 assays are clinically useful methods for HCV RNA quantification and are reliable for predicting the outcome of treatment, despite differences in absolute quantification values and in the correlation between HCV genotypes and viral load. PMID- 10690169 TI - Drug resistance in Campylobacter jejuni, C coli, and C lari isolated from humans in north west England and Wales, 1997. AB - AIMS: To test the sensitivity of strains of Campylobacter species isolated from humans in England and Wales against a range of antimicrobial agents for the purpose of monitoring therapeutic efficacy and as an epidemiological marker. METHODS: An agar dilution breakpoint technique was used to screen isolates against ampicillin, chloramphenicol, gentamicin, kanamycin, neomycin, tetracycline, nalidixic acid, ciprofloxacin, and erythromycin. Minimal inhibitory concentrations (MIC) were also determined for a sample of quinolone resistant strains. RESULTS: Approximately 50% of strains tested were resistant to at least one drug. Strains which were resistant to four or more of the drugs tested were classified as multiresistant; this occurred in 11.3% of C jejuni, 19.9% of C coli, and 63.6% of C lari. Resistance to erythromycin occurred in 1.0% of C jejuni and 12.8% of C coli. Resistance to quinolones occurred in 12% of strains, with a ciprofloxacin MIC of > 8 mg/l and a nalidixic acid MIC of > 256 mg/l; a further 4% of strains had intermediate resistance with a ciprofloxacin MIC of between 0.5 and 2 mg/l (fully sensitive strains, 0.25 mg/l or less) and a nalidixic acid MIC of between 32 and 64 mg/l (fully sensitive strains, 8 mg/l or less). CONCLUSIONS: Resistance to quinolones in campylobacters from human infection may relate to clinical overuse or use of fluoroquinolones in animal husbandry. Both veterinary and clinical use should be reconsidered and fluoroquinolone drugs used only as a treatment for serious infections requiring hospital admission. Erythromycin resistance is still rare in C jejuni but much more common in C coli. PMID- 10690170 TI - The pattern of involvement of the gastric mucosa in lymphocytic gastritis is predictive of the presence of duodenal pathology. AB - AIM: To determine whether the pattern of involvement of the gastric mucosa in lymphocytic gastritis is predictive of the presence or absence of duodenal pathology. METHODS: 50 cases (M:F, 26:24; median age 57 years) diagnosed as lymphocytic gastritis between 1986 and 1998 with concurrent duodenal (D2) biopsies were identified from a computer search of the pathology records and validated by counting gastric intraepithelial lymphocytes. Gastric and duodenal intraepithelial lymphocyte counts were performed on haematoxylin and eosin (H&E) and anti-CD3 stained sections. D2 biopsies were assessed for villous atrophy and chronic inflammatory cell infiltration by subjective grading, and gastritis was classified and graded according to the updated Sydney system. A case was designated corpus predominant when the corpus chronic inflammation grade exceeded that of the antrum. If it was less, then the case was antrum predominant, and if they were equal it was diffuse (pan-) gastritis. The ratio between the corpus and antral intraepithelial lymphocyte count in individual patients was calculated. RESULTS: Of 50 cases of lymphocytic gastritis, 21 were classified as corpus predominant. With one exception (a case of mild villous atrophy), all were accompanied by normal duodenal morphology. Cases with a corpus predominant gastritis had median duodenal intraepithelial lymphocyte counts of 19 (H&E) and 14.1 (CD3), whereas 29 subjects with an antrum predominant or diffuse gastritis had median counts of 39.9 (H&E) and 37.9 (CD3). Fifteen of these 29 cases (52%) showed villous atrophy; all were graded as moderate or severe. Patients with any degree of villous atrophy had a mean corpus/antrum intraepithelial lymphocyte ratio (H&E) of 0.59 (representing antral predominance), while those with normal duodenal morphology had a ratio of 2.39 (p < 0.0001). CONCLUSIONS: The pattern of involvement of gastric mucosa in lymphocytic gastritis is closely related to the associated duodenal pathology. Those with the corpus predominant form are unlikely to have duodenal pathology, while those with an antral predominant or diffuse form should have distal duodenal biopsies taken to exclude villous atrophy. PMID- 10690171 TI - The contribution of MIB 1 in the accurate grading of vulvar intraepithelial neoplasia. AB - AIM: To determine the interobserver variation in scoring presence and grade of vulvar intraepithelial neoplasia (VIN) in haematoxylin/eosin (H/E) slides, MIB 1 slides, and the combined use of H/E and MIB 1 slides. METHODS: 10 slides were stained with H/E and MIB 1 with each of the following diagnoses: normal vulvar skin, VIN 1, VIN 2, and VIN 3. Six observers first scored the H/E slides separately from the MIB 1 slides and second the combined H/E and MIB 1 slides. RESULTS: Unweighted group kappa for MIB 1 was 0.62 and the weighted group kappa was 0.91. This was significantly better than the unweighted group kappa for H/E slides (0.47, p = 0.023) as well as the weighted group kappa for H/E slides (0.82, p = 0.014). There was no improvement by the combined use of H/E and MIB 1 slides. VIN 2 is far less confused with VIN 3 in the combined use of H/E and MIB 1 slides (9%) than in H/E slides (38%) (p = 0.007). There is a tendency to grade VIN in a two tailed grading system rather than a three tailed grading system, which became more apparent with the combined use of H/E and MIB 1 slides. CONCLUSIONS: The interobserver variation with sole use of MIB 1 is better than with the use of H/E stain in VIN. The use of MIB 1 in grading VIN diminishes confusion between VIN 2 and VIN 3 fourfold. A two tailed grading system for VIN seems already to work in daily practice. PMID- 10690172 TI - Effect of using templates on the information included in histopathology reports on specimens of uterine cervix taken by loop excision of the transformation zone. AB - AIM: To determine the change in information relayed from histopathologists to clinicians by using templates for reporting specimens of uterine cervix sampled by loop excision of the transformation zone (LETZ). METHODS: Minimum datasets for the information required from LETZ specimens received from the colposcopy clinic, Royal Infirmary, Edinburgh, were incorporated into templates on the clinical service computer (Pinnacle) of the pathology department, University of Edinburgh. Pathologists completed hard copy versions, which were transcribed into the computer templates for report generation. The effect of the changes on the quality of the pathology reports was studied. The number of cases in which each item in the dataset received comment in template generated reports was compared with that in traditional prose reports compiled before the use of the templates and in prose reports issued after the introduction of the templates. Questionnaire studies were undertaken of clinicians' and pathologists' opinions of the template reports. RESULTS: In the template reports nearly all items received comment in almost 100% of cases. In the prose reports issued both before and after the templates were in use, most items were mentioned in a significantly lower proportion of cases. Clinicians thought the template reports were clearer and the information could be more readily assimilated than from the prose versions. CONCLUSIONS: The use of template reports in these types of specimen allowed more consistent and detailed information transfer. The change appeared to result from the use of the templates rather than from increased awareness of the items to be reported. PMID- 10690173 TI - Papillary hidradenoma: immunohistochemical analysis of steroid receptor profile with a focus on apocrine differentiation. AB - AIM: To make a quantitative evaluation by image analysis of oestrogen receptors, progesterone receptors, and androgen receptors in papillary hidradenomas and anogenital sweat glands. METHODS: 20 papillary hidradenomas and the anogenital sweat glands detected in surgical specimens selected from 10 vulvectomies for squamous carcinoma, eight haemorrhoidectomies, and one anal polypectomy, all from female patients, were investigated by the avidinstreptavidin peroxidase testing system. RESULTS: 90% of papillary hidradenomas and almost all the anogenital sweat glands showed immunoreactivity for oestrogen receptor and, more weakly, for progesterone receptor, with immunolabelled nuclear area ranging from 10% to 90%. Conversely conventional sweat glands did not show any nuclear staining. Overexpression of androgen receptors occurred in 20% of papillary hidradenomas, with nuclear staining strictly bordering papillary epithelium with apocrine differentiation. There was no immunoreactivity for androgen receptors in anogenital sweat glands. CONCLUSIONS: Oestrogen and progesterone receptors seem to represent reliable markers for differentiating between anogenital sweat glands and conventional sweat glands, and a further link to explain why papillary hidradenomas occur almost exclusively in the female anogenital region. Positivity for oestrogen/progesterone receptors suggests that epithelia either of anogenital sweat glands or of papillary hidradenomas are controlled by ovarian steroid hormones. Androgen receptor nuclear staining of the epithelium with apocrine differentiation in vulvar papillary hidradenoma strengthens its homology with breast duct papilloma. PMID- 10690174 TI - Possible involvement of IL-12 expression by Epstein-Barr virus in Sjogren syndrome. AB - AIM: To determine the correlation between interleukin 12 (IL-12) expression and Epstein-Barr virus (EBV) in Sjogren syndrome. METHODS: Indirect immunohistochemical technique, enzyme linked immunosorbent assay (ELISA), and immunoblot analysis were used to investigate IL-12 expression by EBV activation, using 13 surgical specimens and four B cell lines. RESULTS: Marked expression of IL-12 was found in the epithelial cells and the infiltrating B cells of salivary gland tissues from patients with Sjogren syndrome (six of 10 cases), but not in those from normal individuals (none of three cases). A striking topographic correlation between IL-12 and EBV was found. In addition, levels of IL-12 production by B cell lines were clearly enhanced by EBV activation in vitro. CONCLUSIONS: IL-12 expression closely reflects the intracellular event of EBV activation in Sjogren syndrome, and may contribute to the T helper cell type 1 (Th1) cytokine overexpression seen in this disease. PMID- 10690175 TI - Immunohistochemical localisation of androgen receptor in apocrine metaplasia and apocrine adenosis of the breast: relation to oestrogen and progesterone receptors. AB - AIM: To investigate the receptor status of the sex steroid hormones in apocrine metaplasia of the breast. METHODS: 82 cases of apocrine metaplasia, including 18 of the rare lesion apocrine adenosis, were studied immunohistochemically for the expression of androgen receptor, oestrogen receptor, and progesterone receptor proteins on formalin fixed, paraffin embedded tissue sections. The standard avidin biotin complex (ABC) technique was followed and appropriate positive and negative controls were used. RESULTS: All the studied cases (82/82) were positive for androgen receptor, but were negative for oestrogen receptor and progesterone receptor. CONCLUSIONS: Apocrine metaplastic epithelium, unlike the normal breast epithelium, is responsive to androgens, through androgen receptors, rather than to the female sex hormones. This may have clinical implications. PMID- 10690176 TI - A primary care evaluation of three near patient coagulometers. AB - AIM: To compare the reliability and relative costs of three international normalised ratio (INR) near patient tests. MATERIALS: Protime (ITC Technidyne), Coaguchek (Boehringer Mannheim), and TAS (Diagnostic Testing). METHODS: All patients attending one inner city general practice anticoagulation clinic were asked to participate, with two samples provided by patients not taking warfarin. A 5 ml sample of venous whole blood was taken from each patient and a drop immediately added to the prepared Coaguchek test strip followed by the Protime cuvette. The remainder was added to a citrated bottle. A drop of citrated blood was then placed on the TAS test card and the remainder sent to the reference laboratory for analysis. Parallel INR estimation was performed on the different near patient tests at each weekly anticoagulation clinic from July to December 1997. RESULTS: 19 patients receiving long term warfarin treatment provided 62 INR results. INR results ranged from 0.8-8.2 overall and 1.0-5.7 based on the laboratory method. Taking the laboratory method as the gold standard, 12/62 results were < 2.0 and 2/62 were > 4.5. There were no statistical or clinically significant differences between results from the three systems, although all near patient tests showed slightly higher mean readings than the laboratory, and 19 24% of tests would have resulted in different management decisions based on the machine used in comparison with the laboratory INR value. The cost of the near patient test systems varied substantially. CONCLUSIONS: All three near patient test systems are safe and efficient for producing acceptable and reproducible INR results within the therapeutic range in a primary care setting. All the systems were, however, subject to operator dependent variables at the time of blood letting. Adequate training in capillary blood sampling, specific use of the machines, and quality assurance procedures is therefore essential. PMID- 10690177 TI - Cutaneous Scedosporium apiospermum infection in an immunocompromised patient. AB - Scedosporium apiospermum infection occurred in the left forearm of a patient who was taking oral prednisolone for pulmonary fibrosis. The infection appeared to follow a scratch from a blackcurrant bush. This is the first reported case in the United Kingdom of a cutaneous infection from Scedosporium apiospermum in an immunocompromised patient. PMID- 10690178 TI - CD10 positive thyroid marginal zone non-Hodgkin lymphoma. AB - A 72 year old woman presented with swelling of the right lobe of her thyroid gland. Fine needle aspiration and flow cytometry showed a clonal population of B cells expressing CD10 and a diagnosis of follicle centre cell lymphoma was made. Subsequent excision of the thyroid showed the typical histological features of a marginal zone non-Hodgkin lymphoma. Polymerase chain reaction showed no evidence of t (14;18). Immunohistochemistry confirmed CD10 positivity and LN1 (CDw75) expression. This is only the second report of aberrant expression of CD 10 by a marginal zone lymphoma. PMID- 10690179 TI - Deterioration in performance in obtaining bone marrow trephine biopsy cores from children. European Neuroblastoma Study Group. AB - AIM: To complete an audit of bone marrow trephine biopsy adequacy in children MATERIAL: 605 specimens from children with neuroblastoma submitted by 25 centres were reviewed centrally. This reassessment ran between January 1995 and August 1998. RESULTS: 25% of specimens (95% confidence interval (CI) 21% to 29%) were inadequate compared with 17% (95% CI 14% to 20%) in a previous study. Variation between individual centres' performance remains high (5-54% of specimens inadequate). Had five centres performed as well as previously, the inadequate biopsy rate would have been unchanged from that found in the previous study. There was no important improvement in any centre's performance. Earlier suggestions about change in practice have had no discernible impact on centres' ability to obtain adequate bone marrow trephine biopsies from children. CONCLUSIONS: The responsibility for improving the rate of adequate biopsies lies with individual centres. Reporting pathologists might help by making even more positive attempts to influence operators within their own centres. PMID- 10690180 TI - Post-test probability that men in the community with raised plasma ferritin concentrations are hazardous drinkers. AB - BACKGROUND: Raised plasma ferritin concentrations occur unexpectedly during iron studies done by primary care physicians. Plasma ferritin concentration has been positively associated with alcohol use among men. AIM: To determine the post-test probability that men in the community with raised plasma ferritin concentrations are hazardous drinkers. METHODS: The subjects were 152 men, randomly selected from a city's electoral roll. Nineteen (12.5 (2.7)%, mean (SEM)) admitted to drinking hazardously. The pretest probability of a man being a hazardous drinker was 0.125. This was converted to pretest odds of 0.14. The likelihood ratio (the ratio of the probability of obtaining a raised plasma ferritin concentration in a hazardous drinker (sensitivity) to the probability of obtaining a raised plasma ferritin concentration in a non-hazardous drinker (1-specificity)) was calculated for different plasma ferritin cut off points. RESULTS: A plasma ferritin level of > 652 micrograms/l gave the largest likelihood ratio, 4.16. Post-test odds were obtained by multiplying the pretest odds (0.14) by the likelihood ratio (4.16). A plasma ferritin level of > 652 micrograms/l had a post-test odds for a man being a hazardous drinker of 0.58. This was converted to a post-test probability of 0.37. CONCLUSIONS: Inquiries could usefully be made into the alcohol consumption of men with a plasma ferritin concentration > 652 micrograms/l, as approximately one in three would admit to drinking hazardously. PMID- 10690181 TI - Information before coronial necropsy: how much should be available? AB - AIM: To assess the amount and quality of information supplied before undertaking a coroner's necropsy, based on the supposition that insufficient information may adversely affect the quality of the necropsy. METHODS: For a one year period (947 cases), sudden death reports from the coronial jurisdiction of South Yorkshire (West) were audited to assess the quality of information supplied. Seven specific items of information were sought: age, sex, occupation, date of death, location of the body, position of the body, date of last seeing a general practitioner, and relevant medical history. The results from necropsy and non-necropsy cases were compared. RESULTS: Only 22.1% of reports contained all seven items of information. There was no difference between the amount of information supplied in necropsy and non-necropsy cases except about when the general practitioner last saw the deceased. An occupational history was not available in 40.4% of all deaths. CONCLUSIONS: The quality of information supplied to the pathologist before necropsy may be suboptimal and could affect the thoroughness of the necropsy itself. PMID- 10690182 TI - Molecular epidemiology of ocular isolates of adenovirus 8 obtained over nine years. AB - Twenty nine strains of adenovirus 8 have been isolated over nine years in Strasbourg, France, 22 of which were from one private ophthalmologist. To assess a possible relation between these strains, the DNA of adenovirus was analysed by restriction fragment length polymorphism using eight different enzymes. Among these, three proved discriminant (Xba I, Bgl II, Eco RI) and made it possible to define 13 genotypes differing from each other by one to three DNA bands. Seven genotypes were unique isolates, while three, representing 16 strains, were isolated over five to eight years. All the genotypes but one were closely related, with 87% homology. All 13 differed from an adenovirus 8 strain from Lyon (homology 68-76%). This study confirmed the stability of adenovirus 8 in a given population. PMID- 10690183 TI - CD44s as a surrogate marker for distinguishing intraductal papilloma from papillary carcinoma of the breast. AB - BACKGROUND: It has been shown that CD44 variants are differentially expressed in normal and neoplastic breast tissues. The diagnostic value of these markers in distinguishing benign from malignant breast lesions has not been well examined. AIMS: To evaluate the diagnostic value of CD44s in distinguishing between intraductal papillomas and papillary carcinomas of the breast, which may be difficult morphologically. METHODS: Expression of CD44s detected by immunohistochemistry was studied in a series of intraductal papillomas (11) and papillary carcinomas (10). The normal breast tissues surrounding the lesions of these cases served as a control. The number of CD44s positive epithelial cells was scored and categorised as < 10%, 10-70%, or > 70%. RESULTS: Normal breast epithelial cells and all intraductal papillomas (11 of 11) expressed CD44s in a high proportion of cells (> 70%). In contrast, the majority of papillary carcinoma cases (eight of 10) expressed this marker in < 10% of the cells. In the remaining two papillary carcinoma cases, positivity was seen in more than 10% but still less than 70% of the cells. CONCLUSIONS: CD44s detection by immunohistochemistry is useful in distinguishing intraductal papillomas from papillary carcinomas of the breast. PMID- 10690184 TI - Granulocytic sarcoma of the rectum: a rare complication of myelodysplasia. AB - A 67 year old man with myelodysplasia was admitted as an emergency with a six week history of rectal bleeding and diarrhoea. Barium enema showed an irregular polypoid filling defect in the lateral wall of the proximal rectum near the rectosigmoid junction. Histology showed this to be a granulocytic sarcoma (extramedullary granulocytic leukaemia; chloroma) infiltrating the bowel. A low index of suspicion of this lesion results in an incorrect diagnosis in many such cases. A chloroacetate esterase immunoperoxidase stain will confirm the granulocytic nature of the tumour cells. PMID- 10690185 TI - Reporting basal cell carcinoma: a survey of the attitudes of histopathologists. AB - AIMS: To investigate the histopathological reporting of basal cell carcinoma. METHODS: Methods of classification and attitudes to excision margins were ascertained from histopathologists in 130 centres; 82 replies were obtained (63% response rate). RESULTS: 24% of those replying did not use any classification system for basal cell carcinoma. The remainder (76%) used a wide variety of different classification systems. A small number (9%) of those questioned felt reporting on completeness of excision was not important. The majority of histopathologists considered the excision margin was worth reporting but there were differences in methods of processing and reporting biopsies. CONCLUSIONS: There is considerable variation in histopathological reporting of basal cell carcinoma. There is a need for uniformity of histopathological reporting to allow both improved management decisions and comparative audit of this extremely common skin cancer. PMID- 10690186 TI - Effects of sulphapyridine on sperm transport through the rat epididymis and contractility of the epididymal duct. AB - This study was undertaken to investigate the effects of sulphapyridine on the transport of spermatozoa through different regions of the epididymis and on the contractility of the epididymal duct in the rat. Sperm transport was investigated by labelling testicular spermatozoa with [3H]thymidine and measuring intraluminal pressures of the epididymis by micropuncture, using a servo-nulling pressure transducer system. In control rats, the transit times of epididymal spermatozoa from the initial segment to the caput, from the caput to the proximal cauda, and from the proximal cauda to the distal cauda were 2, 6 and 3 days, respectively, giving a total transit time of 11 days. The total transit time was shortened to 8 days after treatment with sulphapyridine at a dosage of 450 mg kg-1 for 38-52 days. The rate of sperm transport was most affected in the caput epididymidis. Measurements of intraluminal pressures showed that sulphapyridine had no effect on spontaneous contractions in any regions of the epididymis. However, the frequency of contraction of the corpus and cauda epididymides in response to administration of 10 micrograms noradrenaline kg-1 in the sulphapyridine-treated rats was significantly higher (P < 0.05) than it was in the controls. Methacholine, at a dose of 20 micrograms kg-1, produced a smaller increase in basal pressure in the caput epididymidis of sulphapyridine-treated rats (P < 0.05) compared with controls. The results led to the conclusion that sulphapyridine increases the rate of sperm transport from the caput through the cauda epididymidis, in part, by changes in the responsiveness of the epididymis to the autonomic nervous system. PMID- 10690187 TI - Effect of oxytocin on concentration of PGF2 alpha in the uterine lumen and subsequent endometrial responsiveness to oxytocin in pigs. AB - The aim of this study was to determine the effect of oxytocin on PGF2 alpha secretion into the uterine lumen of pigs and subsequent endometrial responsiveness to oxytocin in vitro. Cyclic, pregnant and oestradiol-induced pseudopregnant gilts were injected i.v. with vehicle or 20 iu oxytocin 10 min before hysterectomy on day 16 after oestrus. Concentrations of PGF2 alpha and 13,14-dihydro-15-keto PGF2 alpha (PGFM) were significantly increased in uterine flushings collected at hysterectomy (P < 0.05) in pregnant oxytocin-injected gilts. Concentrations of PGF2 alpha and PGFM were greater (P < 0.001) in pregnant than in pseudopregnant and cyclic gilts, and greater (P < 0.01) in pseudopregnant than in cyclic gilts. The ratio of PGFM:PGF2 alpha tended to be greater in cyclic (P < 0.06) and pseudopregnant gilts (P < 0.1) than in pregnant gilts. At 85 +/- 5 min after oxytocin injection, endometrium from each gilt was incubated for 3 h for determination of phosphoinositide hydrolysis and PGF2 alpha secretion in response to treatment with 0 or 100 nmol oxytocin l-1. Endometrial phosphoinositide hydrolysis in response to 100 nmol oxytocin l-1 in vitro was greater (P < 0.05) in cyclic oxytocin-injected gilts than in cyclic vehicle injected gilts. Treatment with oxytocin in vitro did not stimulate phosphoinositide hydrolysis significantly in vehicle- or oxytocin-injected pregnant gilts or pseudopregnant gilts. Endometrial PGF2 alpha secretion increased after treatment with 100 nmol oxytocin l-1 in vitro in cyclic vehicle injected (P < 0.01), cyclic oxytocin-injected (P < 0.01), pregnant vehicle injected (P = 0.06), pseudopregnant vehicle-injected (P < 0.05) and pseudopregnant oxytocin-injected (P < 0.05) gilts, but not in pregnant oxytocin injected gilts. The increase in PGF2 alpha in pseudopregnant oxytocin-injected gilts was less (P < 0.05) than that in cyclic oxytocin-injected gilts. These results indicate that oxytocin increases the concentration of PGF2 alpha and PGFM in the uterine lumen during pregnancy and may upregulate endometrial responsiveness to oxytocin during late dioestrus in pigs, but does not have the latter effect during early pregnancy or oestradiol-induced pseudopregnancy. PMID- 10690188 TI - Effect of transport on pulsatile LH release in ovariectomized ewes with or without prior steroid exposure at different times of year. AB - The initial aim of the present study was to test whether the stress of transport suppresses LH pulsatile secretion in ewes. In a pilot experiment in the late breeding season, transport resulted in an unexpected response in three out of five transported, ovariectomized ewes pretreated with oestradiol and progesterone. Before transport, seasonal suppression of LH pulses had occurred earlier than anticipated, but LH pulsatility suddenly restarted for the period of transport. This finding was reminiscent of unexplained results obtained in ovariectomized ewes infused centrally with high doses of corticotrophin-releasing hormone after pretreatment with low doses of oestradiol with or without progesterone. Hence, an additional aim of the present study was to examine whether these latter results with corticotrophin-releasing hormone could be reproduced by increasing endogenous corticotrophin-releasing hormone secretion by transport. Subsequent experiments used groups of at least eight ovariectomized ewes at different times of the year with or without prior exposure to steroids to assess whether these unexpected observations were associated with season or the prevailing endocrine milieu. In the mid-breeding season, transport for 4 h in the absence of steroid pretreatment for 8 months reduced LH pulse frequency from 7.5 +/- 0.3 to 6.3 +/- 0.4 pulses per 4 h (P < 0.05) and LH pulse amplitude from 2.6 +/- 0.5 to 1.8 +/- 0.3 ng ml-1 (P < 0.05). Similarly, in the mid-breeding season, 34 h after the cessation of pretreatment with oestradiol and progesterone, transport suppressed LH pulse frequency from 6.1 +/- 0.4 to 5.5 +/- 0.3 pulses per 4 h (P < 0.05) with a tendency of effect on amplitude (6.2 +/- 2.7 to 2.61 +/ 0.6 ng ml-1; P = 0.07; note the large variance in the pretransport data). During mid-anoestrus, evidence of a suppressive effect of transport was only observed on LH pulse amplitude (4.7 +/- 0.6 versus 3.0 +/- 0.5 pulses per 4 h; P < 0.05) in ovariectomized ewes that had not been exposed to ovarian steroids for 4 months. Repetition of the pilot experiment with 12 ewes during the transition into anoestrus resulted in one ewe with LH pulses seasonally suppressed but increased by transport; 11 ewes had a distinct pulsatile LH pattern which was decreased by transport in six ewes. In anoestrus, there was no effect of transport on LH pulse frequency or amplitude in intact ewes, or those ovariectomized 2-3 weeks previously, with or without prior oestradiol and progesterone treatment. However, basal concentrations of cortisol were greater in anoestrus than in the breeding season, and the increment in cortisol during transport was similar in anoestrus and the breeding season but greater during the transition into anoestrus (P < 0.05). Progesterone concentrations increased from 0.31 +/- 0.02 ng ml-1 before transport to 0.48 +/- 0.05 ng ml-1 during the second hour of transport (P < 0.05). In conclusion, transport reduced LH pulse frequency and amplitude in ovariectomized ewes that had not been exposed to exogenous steroids for at least 4 months. In most animals, the previously observed increase in LH pulsatility induced by exogenous CRH was not reproduced by increasing endogenous CRH secretion by transport. However, in four ewes, transport did increase LH pulsatility, but only during the transition into anoestrus in ewes with seasonally suppressed LH profiles after withdrawal of steroid pretreatment. PMID- 10690189 TI - Reproductive function in male rats with chronic nephrosis. AB - Endocrine dysfunction has been associated with renal diseases. The present study was conducted to explore reproductive function in male rats with chronic nephrosis. Experimental chronic nephrosis was induced by the administration of 7.5, 5.0 and 5.0 mg per 100 g body weight of puromycin aminonucleoside on days 0, 21 and 35, respectively. Reproductive function was evaluated on the basis of hormonal concentrations, mass of accessory sex organs and fertility during an 84 day period. Circulating LH, FSH, testosterone and oestradiol concentrations were measured by specific radioimmunoassays, while fertility was estimated by the rate of pregnancy induction. Samples were collected on days 7, 14, 28, 56 and 84. The results showed an important endocrine dysfunction characterized by low concentrations of LH and FSH during the first month, after which concentrations were similar to control values or even increased on days 56 and 84. Testosterone and oestradiol decreased significantly at all time points evaluated. The mass of the testes did not alter. However, the mass of the prostate and seminal vesicle decreased only during the first 2 weeks, and became essentially normal thereafter. The reproductive capacity of nephrotic males was eliminated on day 7, whereas on day 14, 16% of the group was able to mate successfully and subsequently most animals recovered their normal reproductive function. This study demonstrates for the first time that rats with experimental chronic nephrosis develop an important endocrine dysfunction, characterized mainly by persistent reduction in testosterone concentrations, which impairs reproductive capacity only transiently. PMID- 10690190 TI - Characterization of endoglin on mouse uterine stromal cells. AB - During the oestrous cycle and early pregnancy, the uterus undergoes a variety of morphological and physiological modifications involving uterine cell proliferation and differentiation as well as extensive tissue remodelling. Transforming growth factor beta (TGF-beta) has powerful effects on these events and thus is thought to have a critical role in uterine physiology. Endoglin is a transmembrane glycoprotein that binds TGF-beta 1 and -beta 3 and interacts with TGF-beta signalling receptors to modulate many effects of this growth factor in different types of cell. Studies in mice revealed the highest concentrations of endoglin in the reproductive tract, notably on stromal cells of cyclic and pregnant uteri. The objective of the present study was to investigate the role of endoglin expressed on uterine stromal cells in binding TGF-beta and in the cellular responses induced by this growth factor. Highly purified populations of uterine stromal cells were isolated by cell affinity to the monoclonal antibody MJ7/18, which is specific to mouse endoglin. Affinity labelling of these cells with 125I-labelled TGF-beta followed by immunoprecipitation with endoglin specific polyclonal 1256:4b antiserum indicated that endoglin expressed at the surface of uterine stromal cells binds TGF-beta 1 and interacts with TGF-beta signalling receptors. Treatment of uterine stromal cells with different concentrations of TGF-beta 1 induced a biphasic proliferative response and addition of MJ7/18 as well as neutralizing TGF-beta antibodies showed endoglin to be a modulator of TGF-beta-induced stromal cell proliferation. Given the importance of TGF-beta in the regulation of uterine physiology, these results indicate a role for endoglin during uterine tissue remodelling and decidualization. PMID- 10690191 TI - Comparison of the steroidogenic capacity of bovine follicular and luteal cells, and corpora lutea originating from dominant follicles of the first or second follicular wave. AB - This study, compared the endocrine function of dominant follicles of the first and second follicular waves (DF1 and DF2, respectively) and the corpora lutea that were subsequently formed. In the experiments conducted in vitro, ovaries were collected from dairy cows on day 6.1 +/- 0.2 or day 14.8 +/- 0.2 of the oestrous cycle to obtain steroidogenically active DF1 (n = 8) and DF2 (n = 7). Granulosa and thecal cells were isolated, dispersed and incubated for 16 h with testosterone (granulosa cells) or forskolin or bLH (thecal cells). Both types of cell were subsequently cultured for 9 days with forskolin and insulin. The viability of the granulosa cells was similar in DF1 and DF2, but the concentration of oestradiol in the follicular fluid was higher in DF1 than in DF2. Production of oestradiol and progesterone by granulosa cells was similar in DF1 and DF2, but androstenedione and progesterone production by thecal cells were 3.5-6.5-fold higher in DF1 than in DF2. During the 9 days of luteinization, progesterone production was similar in DF1- and DF2-derived granulosa cells, but was two- to three-fold higher in DF1- than in DF2-derived thecal cells. Experiments were also conducted in vivo. In Expt 1 in vivo, lactating cows that were assigned to ovulate DF1 or DF2 (n = 9 and 13 in replicate 1 and 2, respectively) were injected with PGF2 alpha on days 6 and 7 or on days 14 and 15 of the oestrous cycle, respectively. A wave by replicate interaction was detected for plasma progesterone concentration in the subsequent cycle: in the first replicate, progesterone production was approximately 40% higher in cows that ovulated DF1; in the second replicate, progesterone production was similar in cows that ovulated DF1 or DF2. In Expt 2, pooled plasma progesterone in the mid luteal phase (days 12-15) after insemination of pregnant and non-pregnant cows was approximately 30% higher in cows that had ovulated DF1 (n = 32) than in cows that had ovulated DF2 (n = 22). This study showed DF1 had a higher steroidogenic capacity compared with DF2, which may be related to the hormonal environment in which the follicles developed. PMID- 10690192 TI - Effect of FSH infusion on follicle development in GnRH agonist-treated gilts. AB - The aim was to investigate the effect of infusion of purified FSH alone on follicle development in hypogonadotrophic GnRH agonist-treated gilts. Large-White hybrid gilts (n = 12) were treated during the mid-luteal phase and again after 28 days (day 0) with a potent slow releasing GnRH agonist. On day 3, seven gilts were infused for 168 h with 1.5 S1 units oFSH h-1 (equivalent to 1.5 units of bioactivity of NIH-FSH-S1 standard) and blood samples were collected. Ovaries were then recovered and all follicles > or = 1 mm in diameter were dissected and incubated for 2 h in 1 ml Eagle's minimum essential medium. The ovaries were recovered from the remaining five GnRH agonist-treated gilts on day 10 and also from five cyclic gilts during the late follicular phase (controls). Plasma FSH concentrations in GnRH agonist-treated gilts were lower (P < 0.01) than in follicular phase controls, increased (P < 0.001) after 1 h of FSH infusion and reached a plateau similar (P > 0.1) to that of controls after 8 h. Basal LH concentrations were similar (P > 0.1) between GnRH agonist-treated and control gilts and remained unchanged (P > 0.1) throughout the infusion period. GnRH agonist treatment reduced (P < 0.01) basal oestradiol concentrations compared with control gilts. Infusion with FSH alone increased (P < 0.001) plasma oestradiol concentrations after 96 h compared with those before infusion; when the animals were killed oestradiol concentrations were higher (P < 0.01) in GnRH agonist-treated gilts infused with FSH than in controls. This was also apparent by vulval swelling and behavioural oestrus. There were more follicles > or 1 mm in diameter in the GnRH agonist-treated groups than in the controls (184, 153 and 86 per animal; P < 0.01). Infusion with FSH increased the maximum follicle diameter (GnRH agonist: < 4 mm; FSH infused: < 12 mm; controls: < 10 mm) and tended to increase (P < 0.07) the mean number of follicles > or = 6 mm diameter per animal (FSH infused: 53; controls: 21). Total oestradiol production in vitro by follicles > or = 1 mm was higher (P < 0.01) in GnRH agonist-treated gilts infused with FSH and in follicular phase controls than in animals treated with GnRH agonist alone. However, oestradiol and testosterone secretion in vitro per follicle > or = 6 mm in diameter was lower (P < 0.05) in FSH-infused animals than in controls. In summary, although infusion of FSH alone stimulated the growth of multiple follicles of preovulatory size in GnRH agonist-treated gilts, steroidogenic output by individual follicles was impaired. PMID- 10690193 TI - Direct effects of ovine follicular fluid on ovarian steroid secretion and expression of markers of cellular differentiation in sheep. AB - The objective of this study was to assess the effect of ovine follicular fluid (FF) treatment (with or without FSH replacement) during the late follicular phase on plasma concentrations of gonadotrophins and the development of the ovulatory follicle. Ovarian steroid secretion and expression of mRNA encoding inhibin alpha and beta A, beta B subunits, P450 aromatase and P450 17 alpha-hydroxylase were used as endpoints. After induction of luteolysis by injection of 100 micrograms cloprostenol on days 10-12, Scottish Blackface ewes were allocated to one of three groups: (1) control (n = 7): no further treatment; (2) FF (n = 9): subcutaneous injections of 3 ml steroid-free ovine follicular fluid at 9 h intervals, 18 and 27 h after cloprostenol injection; (3) FF + FSH (n = 8): injections of follicular fluid as above plus subcutaneous injections of 0.36 iu ovine FSH at 6 h intervals, 18, 24, and 30 h after cloprostenol injection. Jugular venous blood samples were obtained via indwelling cannulae at 6 h intervals from 0 to 36 h after cloprostenol injection, and at 10 min intervals from 12 to 18 h (control phase) and from 30 to 36 h after cloprostenol injection (treatment phase). At laparotomy, 36 h after cloprostenol injection, ovarian venous blood was collected and ovaries were removed and processed for in situ hybridization. Plasma concentrations of FSH, luteinizing hormone (LH) and oestradiol were determined by radioimmunoassay. Follicular fluid treatment resulted in a decrease (P < 0.001) in FSH concentrations associated with an acute decrease in ovarian steroid secretion (P < 0.01) and a specific depression in P450 aromatase, (P < 0.001), inhibin-activin beta B subunit (P < 0.05) and thecal LH receptor (P < 0.001) expression. Follicular fluid treatment had no effect on inhibin-activin alpha and beta A, subunit or P450 17 alpha-hydroxylase expression. FSH co-treatment with follicular fluid restored circulating FSH concentrations to normal values and reversed some of the effects of follicular fluid (androstenedione, testosterone and progesterone secretion, and inhibin beta B and thecal LH receptor expression) but not oestradiol secretion or P450 aromatase expression. It was concluded that the actions of follicular fluid are mediated via both central effects on pituitary FSH secretion and by direct ovarian effects on granulosa cell aromatase activity. The results indicate that follicular fluid contains a factor that inhibits aromatase activity of granulosa cells directly and may play a role in the selection of the dominant follicle. PMID- 10690194 TI - Sources of variation in the morphological characteristics of sperm subpopulations assessed objectively by a novel automated sperm morphology analysis system. AB - There is evidence that the mammalian ejaculate contains distinct subpopulations of spermatozoa and that the variability among these subpopulations may have adaptive and functional significance. This study investigated the precision, reproducibility and operating characteristics of a novel automated sperm morphology analysis system, the Hobson Morphology package, establishing protocols to investigate boar sperm characteristics. Five ejaculates were collected from each of three boars from different genetic lines: Landrace-Meishan introgression, Sireline Large White and Damline Large White. Five semen smears per ejaculate were stained with haematoxylin and eosin. Two hundred spermatozoa per slide were analysed. No significant differences among slides within an ejaculate were detected for sperm tail length (P = 0.770), head width (P = 0.736) and head length (P = 0.615), indicating that both staining and morphology analysis were precise and reproducible. Among the boars, variability in tail length was detected (P = 0.001), but head width (P = 0.114) and length (P = 0.069) did not differ significantly. Multivariate pattern analysis (PATN computer package) highlighted three sub-populations of spermatozoa objectively on the basis of tail length (10.0-22.0 microns, 22.1-73.0 microns and 73.1-130.0 microns). The Landrace-Meishan introgression boar possessed more spermatozoa (P < 0.0001) with tails 73.1-130 microns long. Subsequent analysis of morphology parameters in a pure-bred Meishan boar showed similar measurements for tail length (mean +/- SD; 66.36 +/- 24.70 microns) to the Landrace-Meishan introgression boar (mean +/- SD; 67.09 +/- 21.80 microns). Sperm subpopulations originate during spermatogenesis, when heterogeneous genotypic effects determine the structural features of spermatozoa. The findings of this study confirm that tail length differs between boars and that subpopulations of spermatozoa can be detected within a single ejaculate. PMID- 10690195 TI - Immune cell populations in the equine corpus luteum throughout the oestrous cycle and early pregnancy: an immunohistochemical and flow cytometric study. AB - Recent evidence indicates that the cells of the immune system and their large network of secretory products, or cytokines, play an active role in the ovary throughout the oestrous cycle. In the present study, immune cell populations (T and B lymphocytes, macrophages, granulocytes and eosinophils) and expression of major histocompatibility complex (MHC) class II were investigated in corpora lutea from mares in early (days 2-4), mid- (days 7-10) and late (days 12-14) dioestrus, the post-luteolytic phase (days 16-17) and early pregnancy. The number of T lymphocytes within the corpus luteum increased in the late luteal phase. CD4+ cells did not increase until day 16, whereas the number of CD8+ cells increased before functional luteolysis; an apparently selective luteal infiltration of CD8+ cells was observed. MHC class II expression by non steroidogenic cells was increased in samples from days 16-17, as was the number of infiltrating macrophages. Flow cytometry revealed very low expression of MHC class II by large luteal cells at all stages of the oestrous cycle. In early pregnancy, the number of CD4+ and CD8+ cells and macrophages decreased, as did MHC class II expression, compared with mid-dioestrous samples. B cells were present in very small numbers in all samples examined. Eosinophils were similarly sparsely distributed and numbers decreased further in pregnancy. After exogenous PGF2 alpha administration, populations of CD4+ cells and non-specific esterase staining cells were significantly smaller than after natural luteolysis, whereas eosinophil numbers were increased compared with samples from days 16-17. However, the number of CD8+ and CD5+ cells and MHC class II expression were not significantly different from those observed after natural luteolysis. These findings indicate that populations of immune cells in the equine corpus luteum vary during its lifespan. The selective increase in CD8+ cells before functional luteolysis indicates that they have a physiological role in the regression of the corpus luteum. PMID- 10690196 TI - Differentiation of donor primordial germ cells into functional gametes in the gonads of mixed-sex germline chimaeric chickens produced by transfer of primordial germ cells isolated from embryonic blood. AB - This study was carried out to elucidate whether primordial germ cells, obtained from embryonic blood and transferred into partially sterilized male and female recipient embryos, could differentiate into functional gametes and give rise to viable offspring. Manipulated embryos were cultured until hatching and the chicks were raised until maturity, when they were mated. When the sex of the donor primordial germ cells and the recipient embryo was the same, 15 out of 22 male chimaeric chickens (68.2%) and 10 out of 16 female chimaeric chickens (62.5%) produced donor-derived offspring. When the sex of the donor primordial germ cells and the recipient embryo was different, 4 out of 18 male chimaeric chickens (22.2%) and 2 out of 18 female chimaeric chickens (11.1%) produced donor-derived offspring. The rates of donor-derived offspring from the chimaeric chickens were 0.6-40.0% in male donor and male recipient and 0.4-34.9% in female donor and female recipient. However, the rates of donor-derived offspring from the chimaeric chickens were 0.4-0.9% in male donor and female recipient and 0.1-0.3% in female donor and male recipient. The presence of W chromosome-specific repeating sequences was detected in the sperm samples of male chimaeric chickens produced by transfer of female primordial germ cells. These results indicate that primordial germ cells isolated from embryonic blood can differentiate into functional gametes giving rise to viable offspring in the gonads of opposite-sex recipient embryos and chickens, although the efficiency was very low. PMID- 10690197 TI - Effects of oxytocin and vasopressin on sperm transport from the cauda epididymis in sheep. AB - This study was performed to determine whether oxytocin or vasopressin affect the transport of spermatozoa from the epididymis of rams in vivo. Under general anaesthesia, cannulae were inserted into each ductus deferens and passed into the cauda epididymis of 24 Oxford Down cross rams and the luminal fluid was collected at 10 min intervals for 2-3 h. Animals were divided into seven groups and received either (i) 2 ml 0.9% saline, (ii) 10 micrograms oxytocin, (iii) 100 micrograms oxytocin, (iv) 100 micrograms oxytocin antagonist, (v) 300 micrograms oxytocin antagonist followed by 100 micrograms oxytocin, (vi) 100 micrograms vasopressin, or (vii) 100 micrograms vasopressin followed by 100 micrograms oxytocin, all by i.v. injection. The mass of fluid and number of spermatozoa in each 10 min sample was measured and the motility of the spermatozoa was assessed. Treatment with saline did not affect the mass or the number of spermatozoa in the fluid collected. Oxytocin at 10 micrograms significantly increased both the output of fluid and the number of spermatozoa by twofold. Oxytocin at 100 micrograms produced a greater increase in both fluid output and the number of spermatozoa within 10 min of administration of the peptide. Treatment with oxytocin antagonist had no immediate effect, but subsequently caused a significant reduction in both fluid output and the number of spermatozoa. Pretreatment with oxytocin antagonist inhibited the stimulatory effect of oxytocin. Vasopressin did not increase the number or concentration of spermatozoa in the fluid and appeared to decrease fluid output. No significant changes in the morphology or motility of the spermatozoa collected was observed in any of the samples. These data demonstrate that oxytocin has specific actions on the epididymis to increase sperm transport. They indicate that local oxytocin may be involved in regulating basal contractility of the cauda epididymidis and that augmentation by the peptide in the peripheral circulation, as occurs around the time of ejaculation, may promote a significant increase in the transport of spermatozoa into the vas deferens and ejaculate. PMID- 10690198 TI - Variability in the size of the nucleus in spermatozoa from Houbara bustards, Chlamydotis undulata undulata. AB - Semen collected from 3-year-old male Houbara bustards contained large proportions (6-40%) of spermatozoa with large nuclei. In these spermatozoa, the length of the nucleus was up to twice the mean length of the nucleus in normal spermatozoa. The lengths of the acrosome, midpiece and flagella were all normally distributed, but the length of the nucleus formed a bimodal distribution. The proportion of spermatozoa with large nuclei varied among males, but not among different semen samples collected from the same male throughout the breeding season. The proportion of motile spermatozoa with large nuclei was half that of normal spermatozoa, but their velocity was significantly greater. After insemination into females, spermatozoa with large nuclei were observed in the outer perivitelline layer of eggs laid, indicating that they were stored and transported within the oviduct and reached the egg at about the time of fertilization. Furthermore, there was no difference in the ability to produce viable progeny in females that were mated with males producing greater proportions of spermatozoa with large nuclei compared with those producing 'normal' spermatozoa. Thus, the abnormal spermatozoa did not appear to impede fertility. There were no signs of triploidy in the males that produced spermatozoa with large nuclei, or in their progeny, as demonstrated by the size of erythrocytes. Therefore, it appears that the spermatozoa with large nuclei were the result of aberrant spermatogenesis. PMID- 10690199 TI - Correlation of increased concentration of ovine endometrial cyclooxygenase 2 with the increase in PGE2 and PGD2 in the late luteal phase. AB - Ovine endometrium showed transient expression of high concentrations of the inducible isoform of cyclooxygenase, cyclooxygenase 2 (COX-2), whereas the constitutive isoform, cyclooxygenase 1 (COX-1), was expressed at much lower concentrations and did not change. In this study, the pattern of prostaglandin synthesis in endometrial luminal cells was investigated in relation to their COX 2 content. Endometrial cells from cyclic or pregnant ewes at days 9, 12, 14 and 16 were isolated and analysed for the presence of COX-1 and COX-2 proteins using western blot analysis. Freshly isolated cells were incubated with 0.5 microCi [3H]arachidonic acid ml-1. Radioactive cyclooxygenase metabolites were analysed by reverse-phase HPLC. Luminal cells produced mainly PGF2 alpha, PGE2, PGD2 and 13,14-dihydro-15-keto PGF2 alpha and to a lesser extent 6-keto PGF1 alpha, thromboxane B2 and 13,14-dihydro-15-keto PGE2. The production of PGE2 and PGD2 was proportional to the cellular concentration of COX-2. PGE2 and PGD2 release was low on day 9 when COX-2 was not expressed, whereas high concentrations of PGE2 and PGD2 were synthesized on days 12-14 when COX-2 was highly expressed, reaching 100 ng microgram-1 cellular protein. In contrast, the basal production of PGF2 alpha did not appear to be related to COX-2 concentration and was greatest on day 16. Moreover, the release of PGF2 alpha was maintained at steady state values between days 9 and 14 by the production of 13,14-dihydro-15-keto PGF2 alpha. Although PGF2 alpha output was lower at day 16 of pregnancy compared with the oestrous cycle, no difference was observed in the pattern of prostaglandin synthesis between pregnant and non-pregnant ewes. PMID- 10690200 TI - Sequence of apoptosis and inflammatory necrosis within the formative ovulatory site of sheep follicles. AB - The aim of this study was to define the temporal and spatial patterns of apoptosis, necrosis and inflammation within preovulatory ovine follicles. A gonadotrophin surge was induced in pro-oestrous ewes by GnRH, and isolated follicles were hemisected into apical and basal segments at 0, 10, 18 and 22 h (the time of ovulatory stigma development) after GnRH. Ovarian surface epithelial and granulosa cells were isolated and assessed by fluorescence microscopy for membrane phosphatidylserine translocation-annexin V (early-stage apoptosis), oligonucleosomal DNA nick endlabelling (advanced apoptosis), and nuclear propidium iodide incorporation (necrotic membrane disruption). Thecal shells were analysed for interstitial blood cells. Preovulatory follicles were also hemisected and subjected to electrophoretic DNA degradation analysis. Annexin V binding and in situ DNA fragmentation among ovarian surface epithelial and granulosa cells along the follicular apex were high 18 and 22 h after GnRH. Propidium iodide staining of apical ovarian surface and granulosa cells was apparent at 22 h. There was a coincident increase within the apical theca as the time of ovulation approached in extravasated leucocytes (18 and 22 h) and erythrocytes (22 h). Apoptotic DNA laddering and necrotic DNA smears within the follicular apex were evident on agarose gels at 18 and 22 h, respectively. In contrast, ovarian surface epithelium not associated with the ovulation site and the basal follicular wall were largely unafflicted. It is suggested that both modalities of cellular death, apoptosis and necrosis (with acute inflammation and vascular injury), contribute progressively to follicular stigma formation and ovarian rupture. PMID- 10690201 TI - Effect of exogenous melatonin on neuroendocrine-reproductive function of middle aged female rats. AB - The possible role of melatonin in the regulation of the reproductive system of female rats during ageing was investigated in middle-aged female rats showing irregular duration of the oestrous cycle (n = 30). Blood samples were obtained by jugular venepuncture during the oestrous cycle in control rats. After this experiment was completed, the female rats were treated with melatonin for 2 months and blood samples were obtained at different stages of the oestrous cycle. Plasma LH, FSH and prolactin concentrations were significantly increased in the afternoon of the day of pro-oestrus after melatonin treatment compared with control rats. Moreover, FSH concentrations too were significantly increased on the morning of pro-oestrus and oestrus in melatonin treated rats compared with control rats. Similarly, oestradiol concentrations were significantly higher on the morning of pro-oestrus in melatonin treated rats compared with controls. Another group of rats showing irregular duration of the oestrous cycle was used to study the possible effect of melatonin treatment on the timing of pro-oestrous surges of LH and FSH. The results showed that LH and FSH peak values occurred at 5 h after melatonin treatment. Pituitary responsiveness to LHRH in a 90 min test was also studied in middle-aged rats showing irregular duration of the oestrous cycle that had been injected for 1 month with either melatonin or saline. Prolactin response was unaffected by exogenous melatonin, but a stimulatory effect of melatonin on LH and FSH pituitary responsiveness to LHRH was observed. The results indicate an improved function of the neuroendocrine-reproductive axis in middle-aged rats after melatonin treatment. PMID- 10690202 TI - Single-section counting error when distinguishing between primordial and early primary follicles in sections of rat ovary of different thickness. AB - The number of primordial follicles within an ovary is frequently determined by counting 5, 7 or 10 microns thick sections and multiplying by the fraction of sections counted and a correction factor to adjust for duplicate counts. The objectives of the present study were: (i) to evaluate the accuracy of the correction factor developed by Abercrombie (1946); (ii) to evaluate the accuracy of the classification of primordial follicles from single tissue sections; and (iii) to determine the incorporation rate of 5-bromo-2-deoxyuridine into primordial follicles. In Expt 1, rat ovaries were sectioned at a thickness of 5, 7 or 10 microns. Primordial follicles were counted and classified across ten adjacent ovarian sections. The percentage of primordial follicles from single sections that were counted twice was 10, 9 and 2% in 5, 7 and 10 microns sections, respectively. This was lower than predicted by Abercrombie's method. The major error in counting from single sections was classification of early primary follicles as primordial follicles (55, 33 and 3% in 5, 7 and 10 microns sections, respectively). In Expt 2, a mean of 12 +/- 7% of primordial follicles incorporated 5-bromo-2-deoxyuridine after infusion for 7 days (four of seven rats had no labelled primordial follicles). IN CONCLUSION: (i) Abercrombie's correction factor should not be used for adjusting counts of follicles; (ii) evaluation of primordial follicles from single sections gives inaccurate counts and incorrect classification is of greater importance than duplicate counting, particularly in thinner sections; (iii) for evaluation of the number of follicles, 10 microns is the optimal thickness; and (iv) primordial follicles incorporated 5-bromo-2-deoxyuridine infrequently. PMID- 10690203 TI - Reproductive features of the eastern mole (Scalopus aquaticus) and star-nose mole (Condylura cristata). AB - Since moles are closely related to shrews, the gametes and reproductive tracts of the star-nose mole (Condylura cristata) and the eastern mole (Scalopus aquaticus) were examined to gain further insight into unusual reproductive traits of the Soricidae. Moles display many of these soricid traits, but with some important differences. The cumulus oophorus of Scalopus, ovulated about 16 h after hCG injection, was largely dispersed by hyaluronidase and, though quite dense, was nevertheless more similar to that of higher mammals than to the compact 'ball of the soricid cumulus. Within the female tract in these moles, approximately 85% of the length of the oviduct comprises a narrow ampulla with numerous differentiated crypts that, in shrews, house spermatozoa. However, in contrast to shrews, moles produce considerably larger numbers of spermatozoa, which challenges the proposal that, in shrews, oviductal sperm crypts specifically permit lower sperm production by the males. In the sperm head of these two moles, the acrosome displays the long rostrum that is typical of other Insectivora, and the perforatorium has the barbs by which soricid spermatozoa probably bind to the zona pellucida. Perhaps allied to this, immunoblots indicated that the immunoreactive acrosomal matrix of Scalopus spermatozoa is simpler than the polypeptide complex of the bovine and hamster acrosomal matrix. PMID- 10690204 TI - Examination of the relative role of FSH and LH in the mechanism of ovulatory follicle selection in sheep. AB - The GnRH-antagonist suppression-ovarian autotransplant model (n = 18) was used to examine the relative roles of temporal changes in FSH and LH stimulation on follicle development and selection. Follicle development was stimulated by infusion with oFSH for 3 days and treatments applied for 60 h after progestagen sponge withdrawal and before delivery of an ovulatory stimulus. In Expt 1, there was continuous infusion of FSH with or without small amplitude high frequency LH pulses, or withdrawal of FSH with or without pulsatile LH. In Expt 2, there was acute or gradual withdrawal of FSH at sponge withdrawal with pulsatile LH. The patterns of follicle development and basal and pulsatile ovarian hormone secretion were determined. The maintenance of FSH throughout the artificial follicular phase resulted in multiple follicle development and ovulation (3.3 +/- 0.3). Pulsatile LH stimulated steroid secretion (P < 0.001) but had little effect on ovulation rates (3.8 +/- 0.8) when FSH was maintained. However, withdrawal of FSH in the absence of LH resulted in atresia of the ovulatory follicles and anovulation whereas, when FSH was withdrawn in the presence of LH, preovulatory follicle development was maintained in some animals (3/6 and 5/9 in Expts 1 and 2, respectively) and these ewes had lower (P < 0.05) ovulation rates (1-2 ovulations per ewe). When FSH was withdrawn gradually in the presence of pulsatile LH, 9/9 animals ovulated with ovulation rates in the normal range. These results indicate that ovulatory follicles can transfer their gonadotrophic dependence from FSH to LH. It is hypothesized that the ability of a follicle to respond to this switch in gonadotrophic support is central to the mechanism of follicle selection. PMID- 10690205 TI - Growth and cellular proliferation of pig corpora lutea throughout the oestrous cycle. AB - Corpora lutea were obtained from gilts on days 2, 4, 8, 12, 15 or 18 after oestrus. Luteal fresh masses and DNA contents increased linearly (P < 0.01) from day 2 to day 12 and day 2 to day 15, respectively. Changes in the ratio of protein:DNA were greatest between days 2 and 4 and days 15 and 18, whereas changes in DNA content were relatively small during the same intervals. Thus, a major component of changes in the size of the corpus luteum during the early and late periods of the luteal phase was cellular hypertrophy. Proliferation of luteal cells in vivo (nuclear incorporation of 5-bromo-2-deoxyuridine, a thymidine analogue) was greatest on day 2 and decreased exponentially (P < 0.01) throughout the oestrous cycle. Results from co-localization of 5-bromo-2 deoxyuridine and factor VIII (von Willebrand factor), a marker of endothelial cells, or 5-bromo-2-deoxyuridine and 3 beta-hydroxysteroid dehydrogenase, a marker of steroidogenic cells, indicated that some of the luteal steroidogenic cells proliferated early in luteal development. However, during early and mid cycle, most of the luteal cell proliferation occurred in the endothelial cells. Thus, during growth of the pig corpus luteum, which is extremely rapid, most of the proliferating luteal cells are vascular endothelial cells. This observation is consistent with the high vascularity and blood flow of the mature corpus luteum and implies a critical role for angiogenesis in luteal development in the pig, as has been proposed for several other mammalian species. PMID- 10690206 TI - Inhibition of ovulation by tyrosine kinase inhibitors in the in vitro perfused rat ovary. AB - Protein tyrosine kinase activity, leading to tyrosine phosphorylation of the intracellular domains of receptors or non-receptor proteins, is an important feature of downstream signalling after receptor binding of a variety factors, such as growth factors and cytokines. Since several members of these classes of paracrine-autocrine mediator may be involved in the intraovarian events of ovulation, the present study was designed to evaluate the effect of protein tyrosine kinase inhibition on the in vitro perfused rat ovary. Immature rats were primed with 20 iu pregnant mares' serum gonadotrophin 48 h before surgical isolation of the right ovary with connecting vasculature. The ovary was placed in a perfusion system for either 10 h, to examine ovarian concentrations of the established ovulatory mediators plasminogen activator, prostaglandins E2 and F2 alpha, or for 20 h, enabling a complete ovulatory process to occur in vitro. Ovulation was induced by ovine LH (0.2 microgram ml-1) in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.2 mmol l-1) and the effects of two different protein tyrosine kinase inhibitors, genistein and tyrphostin A25, were studied. Unstimulated control ovaries did not ovulate and showed low secretion of progesterone and oestradiol. Addition of LH + 3-isobutyl 1-methylxanthine resulted in a marked stimulation of steroid release, and ovulations occurred in all ovaries (9.0 +/- 0.9; mean +/- SEM). The protein tyrosine kinase inhibitors, genistein and tyrphostin A25, significantly inhibited ovulation at the higher concentrations tested (3.0 +/- 0.3 at 100 mumol genistein l-1; 5.8 +/- 1.0 at 500 mumol tyrphostin A25 l-1) but no effect was seen at lower concentrations. The presence of genistein and tyrphostin A25 at any concentration used did not significantly decrease the LH + 3-isobutyl-1-methylxanthine-induced progesterone or oestradiol concentrations. The intraovarian concentrations of plasminogen activator activity, and prostaglandin E2 and F2 alpha were not altered by the presence genistein (100 mumol l-1). In conclusion, the results of the present study indicate that protein tyrosine kinase signalling pathways are integral parts of the mammalian ovulatory process but do not involve actions on the synthesis of steroids, plasminogen activator or prostaglandins. PMID- 10690207 TI - Bromocriptine-induced premature oestrus is associated with changes in the pulsatile secretion pattern of follicle-stimulating hormone in beagle bitches. AB - The secretory profiles of LH and FSH were investigated before and during the administration of bromocriptine in six beagle bitches. Plasma samples were obtained via jugular venepuncture at 10 min intervals for 6 h every 2 weeks until the next ovulation. Bromocriptine treatment was started 100 days after ovulation. Both before and after bromocriptine treatment, LH and FSH pulses occurred together. The mean duration of the FSH pulse (120 min) was significantly longer than that of the LH pulse (80 min). The interoestrous interval in the bitches treated with bromocriptine was significantly shorter than that of the preceding cycle (160 +/- 3 versus 206 +/- 24 days). The mean basal plasma FSH concentration (7.4 +/- 0.6 versus 6.1 +/- 0.7 iu l-1) and the mean area under the curve for FSH (46.6 +/- 4.7 versus 40.4 +/- 4.4 iu l-1 in 6 h) increased significantly after the start of the bromocriptine treatment. In contrast, the differences in mean basal plasma LH concentration (2.1 +/- 0.2 versus 2.0 +/- 0.2 micrograms l-1) and the mean area under the curve for LH (19.0 +/- 3.1 versus 19.5 +/- 2.5 micrograms l-1 in 6 h) between the day before and 14 days after the start of the bromocriptine treatment were not significant. Bromocriptine administration also lowered the mean amplitude of the FSH pulse and shortened the mean duration of the FSH pulse, without influencing the LH pulse. In addition to demonstrating the concurrent pulsatile secretion of LH and FSH, the results of the present study demonstrate that the bromocriptine-induced shortening of the interoestrous interval in the bitch is associated with an increase in plasma FSH concentration without a concomitant increase in plasma LH concentration. This finding indicates that treatment with the dopamine agonist bromocriptine increase plasma FSH to a concentration that results in the enhancement of follicle development. PMID- 10690208 TI - Disulfide formation in bovine zona pellucida glycoproteins during fertilization: evidence for the involvement of cystine cross-linkages in hardening of the zona pellucida. AB - The time for solubilization of the bovine zona pellucida in a hypotonic buffer containing 5% (v/v) beta-mercaptoethanol and 7 mol urea l-1 increased by 10% after fertilization. Coupling with a specific fluorescent thiol probe, monobromobimane (mBBr), was markedly greater in the zona pellucida of ovarian eggs compared with fertilized eggs, indicating that the cysteine residues in the zona pellucida of unfertilized eggs are oxidized to cystines during fertilization. After endo-beta-galactosidase digestion to remove N acetyllactosamine repeats of the carbohydrate chains, three zona pellucida glycoproteins (ZPA, ZPB and ZPC) coupled with the fluorescent bimane groups were fractionated efficiently by reverse-phase HPLC. Estimation of bimane groups in the three components and SDS-PAGE revealed that intramolecular disulfide bonds in ZPA and intra- and intermolecular disulfide bonds in ZPB were formed during fertilization, but oxidation of cysteine residues in ZPC was low. Specific proteolysis of ZPA during fertilization was also observed. These results indicate that the formation of disulfide linkages together with specific proteolysis result in the construction of a rigid zona pellucida structure, which is responsible for hardening of the zona pellucida. PMID- 10690209 TI - Sex ratio bias in postpartum-conceived Norway rat litters is produced by embryonic loss in midpregnancy. AB - In rats, dams that conceive in their postpartum oestrus and then lose their firstborn litter bias the sex ratio of the litter toward females in utero. The present study identifies the source of litter sex ratio bias in these postpartum pregnant non-lactating dams. The female bias arises first through the postconception loss of embryos, and second, the loss occurs in midpregnancy between the attachment of the blastocyst to the uterine wall on day 5 and full metrial gland development on day 14. Some pregnancies were restricted to one uterine horn to see if this loss (and thus the opportunity for litter sex ratio biasing) was influenced by local factors operating within the uterine horn. Embryonic loss was more closely associated with the number of embryos implanting in a single horn than with the number implanting in the litter, demonstrating that local crowding within a horn is sufficient for the preferential loss of male embryos. This loss did not cause an obvious decrease in the size of the live-born litter because only those horns with a surfeit of embryos lost them. This process was the same in the right and left horns; both carried and lost the same numbers of embryos. A dam that conceives in her postpartum oestrus and then loses her suckling litter forgoes the implantation delay and uterine healing caused by lactation. Male embryos are less successful at implanting in a uterus only recently vacated by a previous litter. PMID- 10690210 TI - Rotavirus, poliovirus top 1999 immunization changes. PMID- 10690211 TI - Newly approved abacavir to carry hypersensitivity warning. PMID- 10690212 TI - Medicare pays twice the VA cost for drugs, report says. PMID- 10690213 TI - Preparing pharmacy technicians for patient-focused care. PMID- 10690215 TI - Evidence-based medicine for children. PMID- 10690214 TI - St. John's wort: Hypericum perforatum. PMID- 10690216 TI - Stability of rifabutin in two extemporaneously compounded oral liquids. AB - The stability of rifabutin 20 mg/mL in two oral liquids was studied. Powder from 100 150-mg rifabutin capsules was placed in a glass mortar. Cherry syrup (pH 2.9) or a 1:1 mixture of Ora-Sweet and Ora-Plus (Paddock Laboratories) was added to produce 750 mL of each formulation, which was then stored in 2-oz plastic prescription bottles. Three bottles of each formulation were stored at 4, 25, 30, and 40 degrees C. At 0, 1, 2, 4, 8, and 12 weeks, the bottles were collected and allowed to remain at room temperature for one hour; samples of about 1 mL were collected from each bottle, weighed, and assayed for rifabutin content by high performance liquid chromatography. The rifabutin liquids prepared with cherry syrup and stored at 4, 25, and 30 degrees C lost a mean of < 8% of the initial drug concentration during the 12-week study; at 40 degrees C, the liquids lost > 10% of the initial drug concentration by 12 weeks. There was a mean loss of < 5% of the initial rifabutin concentration in all the liquids prepared with Ora-Sweet and Ora-Plus. The liquid prepared with cherry syrup, upon standing, showed a tendency for some of the ingredients to float. The suspension prepared with Ora Sweet and Ora-Plus had a tendency to retain bubbles after it was shaken, but the ingredients did not settle upon standing. Rifabutin 20 mg/mL in two extemporaneously compounded oral liquids prepared from capsules and sweetened vehicles was stable for at least 12 weeks at 4, 25, 30, and 40 degrees C with the exception of rifabutin in cherry syrup, which was stable for only 8 weeks at 40 degrees C. PMID- 10690217 TI - Clinical research: national survey of U.S. pharmacy-based investigational drug services--1997. AB - The results of a survey on the status of pharmacy-based investigational drug services and ways in which some institutions have adapted to recent changes that may affect research are presented. A 99-item survey on investigational drug services was sent in February 1997 to pharmacy directors at 1495 hospitals affiliated with teaching institutions throughout the United States. The survey covered workload, inpatient and outpatient pharmaceutical services, marketing of services, quality assurance, committee involvement, funding sources, computerization, and educational activities. The response rate was 21%; 68% of the respondents were from sites involved in dispensing drugs used for clinical research, with pharmacies at larger hospitals being more likely to dispense investigational drugs than pharmacies at smaller hospitals. The pharmacies participated in a mean 51 active protocols for every full-time-equivalent employee budgeted to research protocols. The correlation between staffing level and number of protocols was high. The service provided most often was the maintenance of drug accountability records for study drugs dispensed to inpatients and outpatients. There was wide variation in the types of services provided to researchers by the pharmacies. Basic services, such as dispensing and inventory control, were provided by nearly all the pharmacies, whereas more specialized services tended to be provided more often at institutions more heavily committed to research. Pharmacy-based investigational drug services at hospitals affiliated with teaching institutions offered basic services, such as dispensing and inventory control, almost universally and offered specialized services to varying degrees. PMID- 10690218 TI - Clinical research: ASHP guidelines and future directions for pharmacists. AB - ASHP guidelines on the pharmacist's role in clinical drug research and future directions for pharmacists in clinical drug research are described. Health-system pharmacists have a responsibility to the patient and to the institution to ensure that clinical research systems are sound, that patients are protected, and that research is conducted in a safe, effective way. ASHP guidelines list minimum standards that are essential for improving performance. The ASHP Guidelines on Clinical Drug Research, approved in November 1997, update information previously found in the ASHP Guidelines for the Use of Investigational Drugs in Organized Health-Care Settings, approved in 1990, but have an expanded focus. Additions include recognition of relevant business practices, implications of technology, and the expansion of clinical research beyond the academic health center. At a minimum, all pharmacists involved in clinical research should handle the record keeping for drug accountability, provide drug information to patients and to other health care providers, ensure the appropriate care of patients at sites not involved in the study, and provide accountability at nonpharmacy locations. Managing and coordinating clinical drug research is an area of growth that represents a great opportunity for clinical drug research. By providing baseline and higher-level pharmaceutical services for clinical research projects, pharmacists can help to ensure data accuracy and completeness and patient safety. PMID- 10690219 TI - ASHP Therapeutic Guidelines on Stress Ulcer Prophylaxis. ASHP Commission on Therapeutics and approved by the ASHP Board of Directors on November 14, 1998. PMID- 10690220 TI - Azithromycin-induced hearing loss. PMID- 10690221 TI - Incompatibility of ceftriaxone sodium with lactated Ringer's injection. PMID- 10690222 TI - [Early detection and intervention of hypoacusia in childhood. Is it a time to change?]. PMID- 10690223 TI - [Ideals, plans and values in adolescence]. PMID- 10690224 TI - [Plan for the early detection and intervention of childhood hypoacusia]. PMID- 10690225 TI - [Testosterone treatment of delayed puberty: a longitudinal study in relation to a control group]. AB - OBJECTIVE: Delayed puberty is a very common clinical situation that affects a great number of adolescents. We analyzed the effects that testosterone therapy produces in this situation, including the start of puberty and, therefore, lessening the psychological effects that this delay causes. PATIENTS AND METHODS: We carried out a longitudinal study, in which we followed the growth and maturation of 32 boys from the age of 14 to 19 years. The sample was divided into a control group (n = 17) and a treatment group (n = 15). The treatment group received 50 mg/month of testosterone enantate depot during 6 months. None of the subjects, neither in the control group nor in the treatment group, had started puberty or if so, they had started it in an insufficient way for their age. RESULTS: The boys treated with testosterone developed a greater growth velocity compared to the control group during the first year of observation (9.07 +/- 1.11 cm/year vs 6.9 +/- 1.76, respectively, p < 0.0001). They had a higher increment in the muscular area of the arm (p < 0.005) and pubertal stage G changes occurred more quickly. On the other hand, the growth of the testicular volume was similar in both groups. At 19 years of age, no significant difference between the groups was observed in any of the clinical parameters studied. CONCLUSIONS: Treatment wit testosterone at the dose used promotes a significant response that leads to the start of puberty, but without stopping the maturation of the hypothalamic pituitary axis that is produced in normal puberty, allowing a normal testicular evolution. The treatment does not show any long-term effects. It is, therefore, an effective treatment of delayed puberty. PMID- 10690226 TI - [Surgical correction of symptomatic ventricular septal defects in patients less than 6 months of age]. AB - OBJECTIVE: Our objective was to evaluate the efficiency of a single surgical intervention in patients with symptomatic interventricular septal defects during the first six months of life. PATIENTS AND METHODS: Between 1989 and 1997, 42 patients, 20 males and 22 females with an average age of 3.9 +/- 0.3 months and an average weight of 4 +/- 0.4 kg, were operated. Seven suffered from Down's syndrome. All of the patients became symptomatic during the first two months of life. The defect was localized by using Echo-Doppler in all of the cases. Thirty six had perimembranous ventricular septal defects, 2 were muscular, 3 multiple and 1 was infundibular. The average defect size was 8 +/- 1.2 mm. A catheter was placed in 34 patient with the following results: Left to right shunt with 2.2 +/- 1.2, right ventricle systolic pressure of 57 +/- 20 mmHg (16 with systemic pulmonary pressure) and an average pulmonary pressure of 38 +/- 1.8 mmHg. The average pulmonary vascular resistance was 28 +/- 1.8 U/m2. Deep hypothermia (18 degrees C was applied during the surgery and the average cardiac arrest time was 31 +/- 4 minutes. RESULTS: None of the patients died during or after the surgical procedure. Patients required minimum ionotropic support during the first hours. The average time in the intensive care unit was 3.5 +/- 0.6 days, with an average hospitalization time of 11.2 +/- 2.1 days. Immediate complications included one hypertensive crisis, four junctional ectopic tachycardias, two atrio-ventricular blocks, 1 transient arrhythmia, two atelectasia-pneumonias, two patients with stridor and two sternal infections. During the follow-up period, two patients required a second intervention to repair the patch. CONCLUSIONS: We believe that one-time surgery is adequate to correct symptomatic ventricular septal defects. PMID- 10690227 TI - [Polydactyly of the hand and foot]. AB - OBJECTIVE: Our aim was to establish the differences between polydactyly of the hand, the foot and those affecting both the hands and feet. PATIENTS AND METHODS: One hundred twenty-five cases of hand polydactyly (HP), 105 of foot polydactyly (FP) and 25 cases of combined hand and foot polydactyly (HFP) were reviewed. We differentiated between preaxial, postaxial, axial and peculiar polydactyly and a group constituted by all other non-preaxial location (OTHERL) was also formed. In all patients the following parameters were analyzed: sex, laterality, antecedence of malformation in the family (FAANT) and the existence of other malformations (OTHERM). RESULTS: Polydactyly was commonly preaxial in the hand (72%), OTHERL in the foot (64.7%) and equally located in hands and feet in HFP, with the most frequent being postaxial/postaxial combination (36% of the cases). Bilaterality is rare in preaxial HP (3.3% versus 77.7% of right unilaterality) and remarkable in OTHERL (54.2%). Bilaterality is greater in preaxial FP (64.8%) and strongly marked in HFP, which accounts for 72% of hands and 80% of feet. There is a slight global dominance of males in all forms. The existence of FAANT is higher in OTHERL in the hand (45.7% versus 34.4% in preaxial) and in preaxial of the foot (45.9% versus 25% in OTHERL), being very high (48%) in HFP. The coexistence with other malformations, either in the hand or foot, is higher in OTHERL. PMID- 10690228 TI - [Reference values for thyroid hormones, thyrotropin and thyroglobulin in healthy children of Zaragoza]. AB - OBJECTIVE: The purpose of this study was to estimate the basal serum concentration reference values for total T3 (T3), total T4 (T4), free T4 (FT4), thyrotropin (TSH) and thyroglobulin (Tg) in healthy children of Zaragoza. PATIENTS AND METHODS: Healthy children aged 0 to 14, with normal weight and height, living in the metropolitan area of Zaragoza (Spain) were the reference population of this transversal study. Basal serum concentrations of T3, T4 and FT4 were measured by radioimmunoassay and of TSH and Tg by immunoradiometric assay. Reference values and ranges were estimated according to the recommendations of the International Federation of Clinical Chemistry. RESULTS: Reference values have been classified according to age, sex and pubertal stage. There are differences in T3, T4, FT4 TSH and Tg concentrations during the prepubertal period according to age, but not to sex, and between the prepubertal period and puberty. Sex and Tanner stage influence T3 and T4 concentrations during puberty. CONCLUSIONS: Since there are differences in T3, T4, FT4, TSH and Tg reference values according to age, sex, pubertal stage and immunoassays, it is necessary to establish reference values for every population and laboratory in accordance with these parameters. PMID- 10690229 TI - [Prevalence of symptoms suggestive of allergic rhinitis and atopic dermatitis in adolescents (Spanish ISAAC Study Group)]. AB - OBJECTIVE: There is a great concern about the worldwide prevalence of allergic disturbances in children and their assumed increase. In order to clarify these facts, the International Study of Asthma and Allergy in Children (ISAAC) group carried out a similar survey in 56 countries, including 463,801 adolescents. We report results of rhinoconjunctival and cutaneous allergies obtained from the 9 centers of the Spanish ISAAC group. PATIENTS AND METHODS: The questionnaire was directly translated from the English text and self-answered by 27,407 schoolchildren between 13-14 years of age from Almeria, Barcelona, Bilbao, Cadiz, Cartagena, Castellon, Pamplona, Valencia and Valladolid. An Epi-Info software was used to assess contingency tables and odds ratios. RESULTS: Only 9.4% of the adolescents said that they had ever suffered hay fever or allergic rhinitis, although 31.3% had nasal symptoms with allergic characteristics during the last 12 months and they were associated with conjunctival disorders in 15.4%. Of these teenagers, 10.3% had ever had atopic dermatitis and 6.2% continued to have active symptoms during the last 12 months. Severe forms were uncommon (0.7%). The female gender had high risk for nasal symptoms during the last 12 months (OR: 1.11; IC 95%: 1.11-1.17) and even higher for atopic dermatitis (OR: 1.3; IC 95%: 1.28 1.37). The frequency of rhinoconjunctivitis and cutaneous disturbances was different in the 9 centers. CONCLUSIONS: We found differences in the prevalence of rhinoconjunctival and cutaneous atopic disorders between centers throughout Spain and now an investigation of the causes is needed. The diagnosis of atopic dermatitis was well recognized by adolescents (10.3%), on the contrary, they frequently (31.3%) reported nasal symptoms with the allergic characteristics but nevertheless do not identify them as such. PMID- 10690230 TI - [Long-term development of children with Schonlein-Henoch purpura associated with anti-neutrophil cytoplasmic antibodies]. AB - OBJECTIVE: The purpose of this study was to assess the frequency of antineutrophil cytoplamic antibodies (ANCA) in Schonlein-Henoch purpura (SHP) and its long-term significance. PATIENTS AND METHODS: IgG and IgA classes of ANCA were studied by indirect immunofluorescence (IIF) and IgG-ANCA against myeloperoxidase (MPO) and against proteinase-3 (PR-3) were determined by ELISA in 50 children with SHP. Eight (16%) of the patients had renal involvement during the acute phase, but none had a permanent nephropathy after a 7-17 year follow up. RESULTS: Positive IgG ANCA were found in 5 (10%) of the cases and only one of these children also had IgA ANCA. The ELISA against MPO and P-3 was negative in all patients. None of the 5 patients with ANCA showed nephropathy during the acute phase nor relapses or permanent nephropathy. CONCLUSIONS: A minority of children with SHP are positive for ANCA and, in the absence of nephropathy, this is not associated with a bad long-term prognosis. PMID- 10690231 TI - [Invasive catheters in neonates]. AB - OBJECTIVE: The care of very sick babies requires the use of invasive catheters in the neonatal intensive care unit. Our objective was to review the invasive catheters placed (umbilical and epicutaneous) between 1994 and 1998 and describe the guidelines used to take care of the intravenous lines. PATIENTS AND METHODS: Two periods were compared (January 1994 until June 1997 and July 1997 until September 1998) and characteristics of the patient and catheter were analyzed. During the first period, sepsis related to the catheter was diagnosed according to clinical and analytical criteria and required a positive blood culture. The same criteria were required in the second period, but coincidence of the organisms in the peripheral and catheter blood culture was also needed. RESULTS: A total of 1,285 central catheters were studied in 958 newborn admissions. Umbilical catheter were used in 6% of the cases and epicutaneous in 23%, most of which were in the upper extremities. The most frequent reason to remove the catheter was the end of the indication. The incidence of catheter related sepsis in the first period was 1% and during the second period 6%. Strict diagnostic criteria used in the second period were more predictive for sepsis. If premature babies were considered alone, the incidence increased to 14%. The most frequent organism isolated was Staphylococcus epidermidis. CONCLUSIONS: To decrease the incidence of sepsis related to catheters, a strict protocol for placement and maintenance must be followed. PMID- 10690232 TI - [Epidemiological study of Steinert's congenital myotonic dystrophy: dysmorphological characteristics]. AB - OBJECTIVE: Steinert's congenital myotonic dystrophy (CMD) is a systemic disease with autosomal mother-to-child transmission and characterized by generalized hypotonia, areflexia, facial diplegia, respiratory and alimentary diseases, arthrogryposis, polyhydramnios, etc. We present the study of the clinical and epidemiological characteristics of Steinert's CMD in our population, with special attention to its dysmorphological features. PATIENTS AND METHODS: In this study we present the analysis of 12 cases of Steinert's CMD identified among 26,956 infants with congenital defects registered by the Spanish Collaborative Study of Congenital Malformations (ECEMC) between April 1976 and March 1998. RESULTS: The minimum estimation of the prevalence in our population is 0.08 per 10,000 live births. We have epidemiologically observed in newborns with Steinert's CMD the presence of a statistically significant difference in the following variables: lower gestational age and birth weight, more polyhydramnios, more feet presentations and Cesarean sections, and a higher frequency of similar congenital defects in first degree relatives. The congenital defects most frequently associated to our population of CMD are located in the extremities, the head and face. CONCLUSIONS: It is important to recognize the congenital defects associated with neuromuscular disorders in the neonatal period, and particularly, the wide spectrum of Steinert's CMD that results in a fetal hypokinesia deformation sequence. PMID- 10690233 TI - [Fibrous hamartoma in children. Report of one case]. PMID- 10690234 TI - [Macrocephaly-cutis marmorata telangectasica congenita: another case of a newly recognized entity]. PMID- 10690235 TI - [Gastric bezoar of cotton swabs]. PMID- 10690236 TI - [Unilateral megalencephaly]. PMID- 10690237 TI - [Intra-thoracic cystic tumors in a newborn]. PMID- 10690238 TI - [Guidelines of basic, advanced and neonatal cardiopulmonary resuscitation (II): basic cardiopulmonary resuscitation in pediatrics. Spanish group of Pediatric and Neonatal Cardiopulmonary Resuscitation]. PMID- 10690239 TI - Should nutrition be considered as a supplementary measure in schistosomiasis control? AB - The most important data on the relationships between nutritional status and schistosomiasis mansoni are reviewed. The probable impact of such findings on the traditional strategies for control of the disease are discussed. In endemic areas, malnutrition and schistosomiasis seem to be associated. Malnutrition impairs the biological development of the parasite. However, like the parasite, it also depresses the host's immune system, and malnutrition and infection can be mutually aggravating. Recent schistosomiasis-control activities, although apparently well designed, have frequently seemed ineffective because of the multiplicity of factors involved, and have not offered a realistic promise of sustainable and definitive control. However, these actions must be continued and even encouraged because they do lead to reductions in the prevalence of infection and, of particular importance, to reductions in the incidence of the more severe forms of the disease. Improvement of the nutritional status of those who inhabit endemic areas, particularly those on low incomes (who are at relatively high risk of malnutrition and of schistosomiasis), is recommended as a supplementary measure. PMID- 10690240 TI - Circulating receptors implicated in the cyto-adherence occurring in severe Plasmodium falciparum malaria in Thailand. AB - The kinetic profiles of soluble chondroitin-sulphate A (CSA), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E selectin were investigated in 17 patients hospitalized with Plasmodium falciparum malaria. The aim was to see if these circulating adhesion molecules could be considered as markers for the severity of P. falciparum malaria. The levels of all the adhesion molecules were found to be higher in the sera from all the malaria cases, both severe and uncomplicated, than in those from uninfected controls. The elevation in plasma CSA, reported for the first time, was statistically very significant (P = 0.00001). However, when severe cases were compared with the uncomplicated, there were no significant differences in the level of any of the receptors except ICAM-1, which was highest in those with the severe disease (P = 0.01). The absence of any significant correlation between the plasma concentration of CSA and malaria severity indicates that this adhesion molecule could not be used to predict the severity of malaria, although its role in sequestration of the parasites in pregnant women is well established. PMID- 10690241 TI - Anaemia in pregnancy: Plasmodium falciparum infection is an important cause in primigravidae in Hoima district, western Uganda. AB - Infection with Plasmodium falciparum is a major cause of anaemia in pregnancy, especially in primigravidae. Of 853 primigravidae visiting an antenatal clinic in Hoima district, western Uganda, for the first time, 530 (62.1%) were found to have P. falciparum parasitaemias and 305 (57.5%) of these had at least 1000 parasites/microliter blood. Plasmodium falciparum parasitaemia was significantly associated with anaemia (relative risk = 0.84, with 95% confidence limits = 0.74 0.96; P = 0.01). Malarial parasites were detected in > 80% of the women who had severe anaemia (P = 0.0008) and haemoglobin concentrations decreased with increasing intensity of infection (P = 0.03). Malarial hyper-reactive splenomegaly was associated with high parasite density (P = 0.01) and low haemoglobin level (P < 0.0001). Effective measures aimed at prevention of malaria and anaemia in pregnancy, especially in primigravidae, would significantly reduce anaemia and its deleterious effects on both the mother and the baby. PMID- 10690242 TI - Re-emergence of Plasmodium malariae in Trinidad, West Indies. AB - A focus of Plasmodium malariae infection has recently occurred on the island of Trinidad, some 30 years after a successful eradication programme. Examination of bloodsmears revealed 22 cases of P. malariae in the Nariva-Mayaro area of Trinidad between August 1994 and September 1995. Most (77%) of the cases were male and, as seven were aged < 25 years of age, it appeared that transmission had been renewed, probably by the vector Anopheles bellator. However, none of the 3000 mosquitoes tested by ELISA for the circumsporozoite protein of P. malariae proved positive. Use of IFAT to check blood samples for P. malariae appeared more sensitive than direct examination of bloodsmears, indicating that 42 (13%) of the 325 samples tested were seropositive (at titres of 1:256 or greater). The levels of transmission of the parasite may therefore be even higher than indicated by examination of bloodsmears. The surveillance measures adopted to understand the epidemiology of this outbreak of P. malariae in Trinidad are described. The need to maintain malaria surveillance in all the countries where P. malariae parasites once existed (prior to eradication) is emphasised. PMID- 10690243 TI - Malaria and the Narmada-river development in India: a case study of the Bargi dam. AB - The largest river-valley development to be proposed in India is that in the Narmada valley. The building of the Bargi dam, a multi-purpose irrigation and hydro-electric project, in Jabalpur, in central India, formed part of the first phase of the development of this valley (1974-1988). Many villages and several hectares of land in three districts were submerged as the waters rose behind the dam, the worst affected area being the catchment area of the primary health centre (PHC) at Narayanganj, in Mandla district. Until recently, cases of malaria were relatively rare in Narayanganj. However, an epidemic of malaria in late 1996 claimed hundreds of lives in the area and the outbreak spread, during 1997, to new villages in the region. A review of the records collected by the National Malaria Eradication Programme (NMEP) not only indicated that the slide positivity rate (SPR) for Narayanganj increased > 7.45-fold between 1979 and 1997 but also that the slide falciparum rate (SFR) increased > 32-fold over the same period. The NMEP data available for Mandla district as a whole indicated a doubling in mean SPR and SFR between 1979 and 1997. There is no evidence that a new species of vector has established since 1979. In fact, indoor-resting densities of anophelines and of the most established vector, Anopheles culicifacies, have fallen since the dam was built, but densities of another vector, An. fluviatilis, have increased. PMID- 10690244 TI - Schistosoma japonicum infection and serum and tissue concentrations of retinol and zinc in pigs. AB - The effects of Schistosoma japonicum infection on the concentrations of zinc in serum, liver, spleen and muscle and on the concentrations of retinol in serum and liver were studied in 48 pigs. Twenty-four of the pigs were each infected by intramuscular inoculation with 2000 cercariae of S. japonicum in medium and the rest were similarly inoculated with parasite-free medium, as controls. On each of weeks 4, 11, 17 and 24 post-inoculation (PI), 12 pigs (six of which were infected) were killed. Tissue samples were collected at necropsy. Blood samples were taken prior to infection and at necropsy from all pigs, and bi-weekly from the pigs killed 24 weeks post-infection. In an analysis of variance in which serum retinol was the dependent variable, the interaction infection x time was found to be significant (P = 0.009). The main reason for this significance was that the concentration of retinol in the sera collected from infected pigs at necropsy at 11 weeks PI was significantly lower than in the control pigs killed at the same time (P = 0.02). Although, overall, infection led to higher zinc concentrations in the liver (P = 0.04) and spleen tissue (P = 0.01), it had no apparent effect on liver retinol, muscle zinc or serum zinc. However, among the pigs which were tested bi-weekly, serum zinc was consistently lower in the infected pigs than in the controls (P = 0.01). The transient declines seen in the concentrations of retinol and zinc in sera from the infected pigs were not accompanied by similar changes in the tissue concentrations, and may reflect an acute-phase response to infection. Schistosoma japonicum infection in pigs is considered a useful model of S. japonicum infection (and probably also of S. mansoni infection) in humans. Similar effects, if they occur in the human infections, may lead to misclassification of vitamin-A and zinc status in endemic populations if this status is based on serum retinol and serum zinc. PMID- 10690245 TI - Efficacy of albendazole and its combinations with ivermectin or diethylcarbamazine (DEC) in the treatment of Trichuris trichiura infections in Sri Lanka. AB - The efficacy of the drugs currently available for treatment of infection with Trichuris trichiura is low compared with that of the drugs used against roundworm and hookworm. Single-dose combinations of albendazole with ivermectin or of albendazole with diethylcarbamazine (DEC) have recently been seen to produce raid and sustained reductions in Wuchereria bancrofti microfilaraemia. This observation prompted the present study, on the efficacy of these combinations against trichuriasis. The drug regimens tested were albendazole (400 mg) alone, albendazole (400 mg) with ivermectin (200 micrograms/kg), and albendazole (400 mg) with DEC (6 mg/kg). Most (155) of the 176 children (4-14 years of age) who each provided a single, pre-treatment, stool sample were found positive for Trichuris ova. These 155 were each randomly allocated to one of the three treatment groups and checked for infection 3 weeks post-treatment, again by a single stool examination. Single-dose therapy with albendazole plus ivermectin produced a 'cure rate' (79.3%) and an egg-reduction rate (93.8%) which were significantly higher than the corresponding rates produced by albendazole alone or albendazole plus DEC (P < 0.01 for each). The efficacies of albendazole with DEC and of albendazole alone were statistically equivalent. Single-dose treatment with the albendazole-ivermectin combination appears to be highly effective against trichuriasis and could prove valuable for routine use. PMID- 10690246 TI - Preventive immunisation could reduce the risk of meningococcal epidemics in the African meningitis belt. AB - Control of meningitis epidemics is based on early case detection followed by mass campaigns of immunisation. However, this strategy showed severe inadequacies during recent outbreaks in Africa. In Niamey, Niger, meningococcal vaccinations began in 1978 and detailed bacteriological and epidemiological surveillance of meningitis started in 1981. When vaccine coverage rates were higher than 50%, the prevalences of Neisseria meningitidis A meningitis were low in Niamey, although there was a concurrent epidemic in rural Niger. A massive outbreak of meningitis in Niamey in 1994-1995 followed a 6-year period during which the mean rate of vaccine coverage remained < 25%. The data indicate that, in the meningitis belt, preventive immunization should avoid a great number of deaths and be less expensive than mass immunisation campaigns performed after epidemics have begun. PMID- 10690247 TI - The molluscicidal properties of Apodytes dimidiata (Icacinaceae): geographical variation in molluscicidal potency. AB - Synthetic molluscicides have proved too expensive for most countries wanting to include snail control in their anti-schistosomiasis programmes. An alternative, which is not only cheaper but also promotes self-reliance and empowerment of the affected communities, is the use of molluscicidal plants. An often-cited limitation to using such natural products is the geographical variation in the toxicity of candidate species. The geographical variation in the molluscicidal activity of Apodytes dimidiata was investigated in South Africa. Leaves of this plant were collected from six, widely separate localities within the areas of the country where schistosomiasis is endemic. The results of bio-assays using the intermediate host snail, Bulinus africanus, clearly showed that variation in toxicity did exist and appeared to be correlated with the range in mean annual temperatures and altitude where the plants grew. Whether the variation was a phenoplastic response to the environment or genetically determined still has to be investigated. Nevertheless, a thorough knowledge of the geographical variation in the level of the active compound(s) in the candidate plants in endemic areas will be needed prior to the implementation of plant-propagation and snail-control programmes. PMID- 10690248 TI - Gross histopathological effects of an extract of Agave attenuata on the epithelium of the digestive tract of Bulinus africanus. AB - A lethal concentration of a crude, aqueous extract of Agave attenuata was applied as a contact poison to Bulinus africanus, the intermediate host of Schistosoma haematobium, for a 24-h period. The gross histopathological effects of the extract on the epithelium of the digestive tract were then studied. A graded series of cellular injuries to the epithelial layer was observed along the length of the tract. These included the loss of cilia and brush border, disruption of the epithelial layer, cellular vacuolation, swelling and rupture, and the discharge of secretory products from mucous gland cells. The results of the microscopy show that epithelial tissue is probably a primary target of the molluscicide. The cytological injuries induced by extracts of A. attenuata indicate that the molluscicide acts by disrupting the osmoregulatory mechanisms of the epithelial cells, but further, detailed studies are required to confirm this. PMID- 10690249 TI - A prospective study of combination antimalarial therapy and of anaemia in Zambian children with cerebral malaria. PMID- 10690250 TI - Plasmodium falciparum infection following allogeneic bone-marrow transplantation. PMID- 10690251 TI - High seropositivity against listeriolysin O in humans. PMID- 10690252 TI - Measles: a disease that has to be eradicated. AB - The incidence of measles is on the decline but it still claims the lives of one million children annually worldwide. The devastating effects of the disease on the health and nutrition of children in developing countries and its high mortality are well documented. The rapid decay of maternal antibodies in infants in developing countries results in early susceptibility to the disease and hence the general recommendation to vaccinate at the age of 9 months. Sustained international efforts have raised global vaccination coverage rates to around 80% at which level it has remained static. Many countries in the western hemisphere have eliminated the disease by adopting aggressive strategies, which include one off 'catch-up' mass campaigns to vaccinate all children aged 1-14 years, 'mop-up' campaigns targeting children who were missed during the 'catch-up' campaign, efficient routine vaccination services capable of reaching 90% of infants, strong surveillance activities, prompt outbreak response, and 'follow-up' campaigns every 2-4 years which target 1-4-year-old children. This success story coupled with the fact that measles has many biological features favouring eradication, and considering that it is a severe and lethal disease still prevailing in many areas, calls for immediate international adoption of eradication goals within a specified period of time. PMID- 10690253 TI - Paediatric hospital admissions at a South African urban regional hospital: the impact of HIV, 1992-1997. AB - Rates of infection by the human immunodeficiency virus (HIV) have been increasing rapidly in South Africa over the last decade. This study documents the changes over time in prevalence of HIV infection amongst hospitalized children, and its effects on the profile of disease and in-hospital mortality over the period 1992 1997. Admissions to the paediatric medical wards between January 1992 and April 1997 were obtained from the routine computerized database held in the Department of Paediatrics at Chris Hani Baragwanath Hospital. HIV tests were performed on clinical indications only. Over the study period there were 22,633 admissions involving 19,918 children. Total annual admissions increased by 23.6% between 1992 and 1996. Prevalence of HIV infection increased from 2.9% in 1992 to 20% in 1997. HIV-infected children had a younger age distribution, longer median length of stay and more readmissions (p < 0.001) compared with HIV-negative and untested children. HIV-infected children accounted for the increased number of admissions for pneumonia, gastro-enteritis, malnutrition and tuberculosis, and the rise in in-hospital mortality by 42% from 4.3% in 1992 to 6.1% in 1997. Paediatric HIV infection has changed the profile of paediatric admission diagnoses and increased in-hospital mortality in the relatively short time between 1992 and 1997. Over the same period, HIV-negative children showed declining rates of malnutrition, vaccine-preventable diseases and admission to the intensive care unit. PMID- 10690254 TI - Knowledge of and attitudes to HIV/AIDS of senior secondary school pupils and trainee teachers in Udupi District, Karnataka, India. AB - A cross-sectional descriptive study using a questionnaire with mostly closed ended questions was carried out on 990 pupils and 46 trainee teachers to investigate their knowledge of and attitudes to HIV/AIDS. Pupils in one school were reassessed after a health talk and distribution of a handout. Despite having had no formal sex education, most respondents were reasonably well informed about the transmission of HIV. However, there were many misconceptions about transmission and prevention and 16.9% of pupils were found to possess very little knowledge of HIV/AIDS. Mass media, teachers and health workers were quoted as the main sources of knowledge. It was found that 24.3% pupils and 6.3% of trainee teachers thought there was a cure, and 27.4% of pupils and 14% of trainee teachers thought there was a vaccine to prevent HIV infection. Schools that were rural, private and English-speaking scored better, as did male students and schools teaching science. The necessity of formal sex education was expressed by 98.5% of pupils and all the trainee teachers. The pupils who were reassessed after receiving a talk and handout showed significant improvement in their knowledge and a change in attitude (p < 0.01). The mass media are important in disseminating knowledge on HIV/AIDS in India but due to the lack of inter personal approaches to the education system, knowledge is inadequate and misconceptions exist. PMID- 10690255 TI - Risk factors for an adverse outcome in bacterial meningitis in the tropics: a reappraisal with focus on the significance and risk of seizures. AB - The relationship of presentation to outcome in children with meningitis was analysed. The relative risk (95% confidence interval) of an adverse outcome (death or neurological sequelae) associated with presentation with at least three of ten features (age < or = 2 yrs, ill for > 7 days, antibiotic treatment, focal nerve deficits, abnormal posturing, abnormal muscle tone, lack of typical meningeal signs, shock, unrousable coma and seizures) was 4.9 (2.7, 8.8), p < 0.0001. The first six features were particularly associated with neurological sequelae, and shock and coma with death. Seizures were associated with either outcome. Two seizure types could be distinguished: seizures which occurred before or on diagnosis only (type I seizures) and seizures which occurred before and/or after diagnosis (type II seizures). Death occurred in 0/41 children without seizures and in 14/34 and 11/34 children with type I and type II seizures, respectively (p < 0.0001). Neurological sequelae occurred in 3/42 children without seizures and in 5/20 and 14/23 with type I and type II seizures, respectively (p < 0.0001). PMID- 10690256 TI - Sydenham's chorea: risk factors and the role of prophylactic benzathine penicillin G in preventing recurrence. AB - To determine the effect of prophylactic long-acting penicillin G in preventing recurrence of Sydenham's chorea and to discover the risk factors associated with occurrence of symptoms, 18 children with symptoms over a 5-year period were prospectively identified. Of these, ten were boys and eight were girls. The majority occurred between the ages of 8 and 10 years [mean (SD) 9.10 (2.62) years]. Sydenham's chorea was generalized in 14 children and one-sided in four. There was no difference in the incidence of right- and left-sided hemichorea. Among the risk factors examined, only a history of chorea in relatives had a significant association with the occurrence of Sydenham's chorea (OR = 6.39; 95% CI 1.30-31.3). A comparison of recurrence between those given prophylactic long acting penicillin G and those who had none showed a statistically significant difference in the recurrence experience between the two groups (p < 0.02). PMID- 10690257 TI - Intravenous immunoglobulin in very severe childhood Guillain-Barre syndrome. AB - To evaluate intravenous immunoglobulin (IVIG) therapy in children with very severe Guilain-Barre syndrome (GBS) with reference to the need for respiratory support, ICU stay and long-term outcome, we studied 33 children with very severe GBS and quadriparesis and/or respiratory muscle weakness admitted to the Pediatric Intensive Care Unit (PICU) of PGIMER, Chandigarh. Cases (n = 22, IVIG group) were enrolled prospectively, and controls (n = 11), similar to cases in age and severity of illness, retrospectively. All children received similar supportive and respiratory care. In addition, cases were given IVIG (Sandoglobulin, Sandoz) 0.4 g/kg bodyweight per day for 5 days. The mean age, duration of symptoms prior to admission and severity of illness in the two groups were similar. In the IVIG group, onset of recovery of muscle power was significantly earlier (day 14.8 (6.8) of illness vs day 20.9 (8.6), p < 0.05) and the length of PICU stay significantly shorter (20.5 (13.0) days vs 50.5 (33.3) days, p < 0.01). Sixteen (72.7%) children in the IVIG group had improved by at least one functional grade after 1 month and 15 (68%) were walking independently after 3 months compared with two (18%) and four (36%) controls, respectively (p < 0.05). The number of children who needed endotracheal intubation and mechanical ventilation and the duration of mechanical ventilation was significantly less in the IVIG-treated group. We conclude that in very severe GBS in children IVIG therapy improves outcome to a remarkable extent, reduces the need for intubation and mechanical ventilation, shortens the length of stay in ICU, and promotes ambulation sooner. PMID- 10690258 TI - Serum levels of measles IgG antibody activity in children under 5 years in Dar-es Salaam, Tanzania. AB - Measles IgG antibody levels were estimated in sera from 685 Tanzanian children, 374 (54.6%) boys and 311 (45.4%) girls aged 18 months to 5 years, using an enzyme linked immunosorbent assay (ELISA). The children were screened for HIV-1 and 2 antibodies using ELISA, and reactive sera were confirmed by Western blot. Nutritional status was assessed by anthropometry. Overall measles vaccination coverage was 98.8%. Measles antibody activity was not detected in 41 (6.0%) children, and ten (1.5%) had antibody levels below 200 mIU/ml, the cut-off level considered to be protective. The non-reactive samples were from one unvaccinated child, one child with unknown vaccination status and 39 vaccinated children. Measles IgG antibody levels were higher in girls (3452.1 mIU/ml) than in boys (2928.2 mIU/ml) (p = 0.02). Higher mean levels were found in children with a history of low birthweight (< 2.5 kg) (p = 0.03). There were no significant differences in measles antibody levels with regard to variations in nutritional status. No correlation (r2 = 0.002) was found between antibody levels and time elapsed since vaccination. In a multivariable logistic regression analysis, children who were HIV-seropositive (n = 9) were more likely to have non protective antibody levels < 200 mIU/ml (OR = 5.85; 95% CI: 1.37-24.93). PMID- 10690259 TI - Comparative study of autonomic nervous system activity in malnourished and normal children in India. AB - Information on the relationship between autonomic functions and malnutrition in children is scant. In the present study, autonomic function tests were conducted in 30 normal subjects and 30 malnourished children aged between 5 and 10 years. The tests performed included tests for parasympathetic functions (resting heart rate, standing-to-lying ratio, lying-to-standing ratio and Valsalva ratio) and tests to assess sympathetic function (hand grip test, galvanic skin resistance). The malnourished children had significantly lower mean weights-for-age (-2.6 Z vs -1.5 Z; p = 0.001), heights-for-age (-2.5 Z vs -1.5 Z; p = 0.001) and weights-for height (-1.6 Z vs -0.8 Z; p = 0.001). Parasympathetic function tests evaluated were significantly affected in malnourished children. Resting heart rate was significantly higher in the malnourished group (90.6 vs 82.5/min; p = 0.001). The other parasympathetic function tests had significantly lower mean values than in the control group, namely, standing-to-lying ratio (1.25 vs 1.32; p = 0.026), lying-to-standing ratio (1.23 vs 1.29; p = 0.021) and Valsalva ratio (1.26 vs 1.28; p = 0.037). Of the sympathetic function tests conducted, there were no differences between the two groups for hand grip test but galvanic skin resistance was significantly higher in the malnourished subjects (190.1 vs 149.73; p = 0.001). It is concluded that autonomic nervous system function is significantly compromised in malnourished children. PMID- 10690260 TI - Hydatid disease of the liver in children. AB - Between 1986 and 1997, 21 children (ten boys and 11 girls) had surgery for hydatid disease of the liver. Their mean age was 6.5 years (range 3-12). Abdominal distention with a mass was the commonest presenting symptom (71.4%), followed by abdominal pain (38%). Hepatomegaly with a palpable mass was present in 12 (57%). Three children had concomitant pulmonary and brain hydatid disease. The diagnosis was established clinically and by skin testing, serology and imaging techniques. All patients received a pre-operative course of mebendazole (50 mg/kg/day) for between 1 and 8 weeks. At surgery, 11 children had a single cyst, eight of which were in the right lobe of the liver. Ten children had multiple cysts occupying both liver lobes. Three forms of surgical treatment were used: capitonnage + partial excision of fibrous capsule; total excision of the cyst; and external drainage of the cyst cavity. Three children required re operation. Mean follow-up time was 24 months. There were no deaths, but five children developed post-operative complications. Surgical treatment in the form of primary closure of the cyst cavity without drainage seems to offer the best therapeutic option for patients with large hydatid cysts. PMID- 10690261 TI - Sotos syndrome (cerebral gigantism): a clinical and radiological study of 14 cases from Saudi Arabia. AB - Fourteen children (of Arab ethnic origin) with Sotos syndrome are described. They were referred to King Khalid University Hospital, Riyadh between July 1992 and June 1997. Their phenotypic characteristics were compared with established diagnostic criteria. There was a male:female ratio of 1.3:1 and a high rate of consanguinity (36%) among parents. At birth, 54% were large and about one-third showed increased height and occipitofrontal head circumference (OFHC). The neonatal histories revealed respiratory and feeding problems in 21%, followed later by delayed motor milestones and speech development in 57%. During childhood, weight, height and OFHC increased further to > 97th centile in 71%, 71% and 93%, respectively. A seizure disorder affected 43%, and 75% had mental retardation (IQ < 70). A non-specific EEG abnormality was found in half of those with seizures. Cranial CT/MRI showed ventricular dilatation in 15% and one patient had corpus callosum dysgenesis. Abdominal ultrasound revealed hydronephrosis in two patients. Radiological cephalometric measurements showed relative prognathism in cases of Sotos syndrome compared with controls (p = 0.003). The study highlights the importance of considering Sotos syndrome in children who present with psychomotor delay. PMID- 10690262 TI - Cholecystitis in children in Zaria, Nigeria. AB - Much has been published on cholecystitis in childhood from Europe and North America but reports from tropical Africa are few. This is a report of seven children with cholecystitis seen over a 10-year period in Zaria, northern Nigeria. Six of the children had acalculous cholecystitis but a predisposing condition (adenomatous hyperplasia) was identifiable in only one. Five presented with complications (perforation, three; gangrene, one; empyema, one). One child without haemolytic disease had calculous cholecystitis and choledocholithiasis. Pre-operative diagnosis of cholecystitis was made in only three cases. Cholecystectomy, two as interval procedures, was safely performed in all patients. In environments such as ours, where cholecystitis in childhood is uncommon, awareness and a high index of suspicion are necessary for early diagnosis and prompt treatment in order to avoid potentially life-threatening complications. PMID- 10690263 TI - Salmonella cholecystitis in a neonate. PMID- 10690264 TI - Benign recurrent intrahepatic cholestasis in a Saudi child. AB - We report a case of benign recurrent intrahepatic cholestasis (BRIC) in an 11 year-old Saudi girl who developed three episodes of pruritus and jaundice at the ages of 4, 8, and 9 years. These episodes were almost stereotypic and lasted 5-8 weeks. Although she had elevated liver enzymes and serum bile acids in her blood during the attacks, they returned to normal between attacks. Thorough investigation excluded other causes of liver disease and her recurrent attacks were shortened by cholestyramine therapy. A diagnosis of BRIC should be kept in mind in patients with cholestasis. PMID- 10690265 TI - Influence of fibrinogen and haematocrit on erythrocyte sedimentation kinetics. AB - This study investigates the influence of haematocrit, fibrinogen concentration and fibrinogen availability (amount of fibrinogen per red blood cell) on erythrocyte sedimentation. The Westergren technique was applied to blood samples from 36 subjects and to their blood manipulated to haematocrits of 10, 20, 30 and 40%. Readings were taken every 10 minutes for 300 minutes. Previous studies indicate that erythrocyte sedimentation occurs in three phases. In this study, we show that haematocrit has little influence on either the rate of fall of particles in the first phase (m1) or the duration of the first phase. This is also true for fibrinogen availability and for fibrinogen concentration at low haematocrits. At high haematocrits m1 increases with fibrinogen concentration. The rate of fall of rouleaux during phase 2 (m2) and ESR60 both decrease exponentially with haematocrit and increase linearly with fibrinogen concentration. While m2 is more closely correlated to fibrinogen availability than to fibrinogen concentration or to haematocrit, this is not the case for ESR60. Thus haematocrit, fibrinogen concentration and fibrinogen availability are more important to the velocity of sedimentation in the second phase than to the sedimenting velocity during phase 1 or to the duration of phase 1. PMID- 10690266 TI - Distributions of rheological parameters in populations of human erythrocytes. AB - We have previously proposed the osmofiltration method based on a modified Hanss hemorheometer to analyze distributions of erythrocytes in their ability to pass through membrane filters with 3 microns pores. Upon decrease in medium osmolality (u) the erythrocyte volume increases. When cell volume becomes V = Vcr at u = ucr, such cell loses its ability to pass through a 3 microns pore. The flow rate of erythrocyte suspension containing cells with different ucr through a filter gradually decreases with decreasing medium osmolality. This rate becomes zero at some u = omega, when the number of non-filterable cells in the applied sample approaches the number of pores in filter. Experimental determination of the dependencies of the filtration rate on medium osmolality for various hematocrit values allows to obtain omega for each hematocrit and, thereby, to assess the distribution of erythrocytes in ucr. Here, we propose a simplified version of this method, which allows screening of the erythrocytes in heterogeneous suspensions for the distribution in ucr by measuring omega for only two hematocrit values, 0.1% and 1%. Applications of the proposed method are exemplified by analysing the erythrocyte populations of healthy donors, of patients with microspherocytosis, hemochromatosis and normal erythrocyte populations in an acidic environment. PMID- 10690267 TI - A new capillary viscometer for whole blood viscosimetry. AB - A new capillary system was developed, incorporating infrared sensors, which allowed the determination of whole blood viscosity over a wide range of shear stresses. Flow conditions were defined by the geometry of the capillary and the sample pressure head. Whole blood was considered to be a power law fluid and a modified Mooney's formula was used for the calculation of the related invariants. The new viscometer proved to be very simple in use, requiring one run, had a short measuring time and utilised a small test sample volume. However it can be used for whole blood viscosity measurements only at medium and high shear stresses. PMID- 10690268 TI - Mucus liquid crystallinity: is function related to microstructural domain size? AB - The size of liquid crystalline domains formed in partially dried giraffe saliva is found to be an order of magnitude greater than that previously documented for slug pedal mucus. A correlation between (a) intrinsic liquid crystalline domain size and (b) the scale of surface topography over which the mucus is required to provide lubrication is postulated. The scale of mucus microstructure can be related, via a simple model, to two significant material constants: the elastic constant K for distortion of molecular alignment in the liquid crystalline state, and the anchoring energy I at the liquid crystal/substrate interface. In turn, the quantity K is expected to depend on fundamental molecular characteristics of the constituent mucin, such as molecular weight and degree of glycosylation. The possibility that observations of mucus microstructure might serve as a diagnostic tool for mucus defects at the molecular level is suggested. PMID- 10690269 TI - Theoretical study on flow-dependent concentration polarization of low density lipoproteins at the luminal surface of a straight artery. AB - It is suspected that physical and fluid mechanical factors play important roles in the localization of atherosclerotic lesions and intimal hyperplasia in man by affecting the transport of cholesterol in flowing blood to arterial walls. Hence, we have studied theoretically the effects of various physical and fluid mechanical factors such as wall shear rate, diffusivity of low density lipoproteins (LDL), and filtration velocity of water at the vessel wall on surface concentration of LDL at an arterial wall by means of a computer simulation of convective and diffusive transport of LDL in flowing blood to the wall of a straight artery under conditions of a steady flow. It was found that under normal physiologic conditions prevailing in the human arterial system, due to the presence of a filtration flow of water at the vessel wall, flow-dependent concentration polarization (accumulation or depletion) of LDL occurs at a blood/endothelium boundary. The surface concentration of LDL at an arterial wall takes higher values than that in the bulk flow in that vessel, and it is affected by three major factors, that is, wall shear rate, gamma w, filtration velocity of water at the vessel wall, Vw, and the distance from the entrance of the artery, L. It increases with increasing Vw and L, and decreasing gamma w hence the flow rate. Thus, under certain circumstances, the surface concentration of LDL could rise locally to a value which is several times higher than that in the bulk flow, or drop locally to a value even lower than a critical concentration for the maintenance of normal functions and survival of cells forming the vessel wall. These results suggest the possibility that all the vascular phenomena such as the localization of atherosclerotic lesions and intimal hyperplasia, formation of cerebral aneurysms, and adaptive changes of lumen diameter and wall structure of arteries and veins to certain changes in hemodynamic conditions in the circulation are governed by this flow-dependent concentration polarization of LDL which carry cholesterol. PMID- 10690270 TI - Flow-dependent concentration polarization of plasma proteins at the luminal surface of a cultured endothelial cell monolayer. AB - Flow-dependent concentration or depletion of atherogenic low density lipoproteins which has been theoretically predicted to occur at a blood/endothelium boundary may play an important role in the genesis, progression, and regression of atherosclerosis in man and intimal hyperplasia in vascular grafts implanted in the arterial system in man and experimental animals. Hence to explore such a possibility, we have studied the effect of a steady shear flow on concentration polarization of plasma proteins and lipoproteins at the luminal surface of a cultured bovine aortic endothelial cell (BAEC) monolayer which served as a model of the vessel wall of an artery or an implanted vascular graft. The study was carried out by circulating a cell culture medium containing fetal calf serum or bovine plasma lipoproteins in steady flow through a parallel-plate flow cell in which a cultured BAEC monolayer was installed, over the physiologic ranges of wall shear rate and water filtration velocity at the BAEC monolayer. The water (cell culture medium) filtration velocity at the BAEC monolayer was determined to provide a measure of the change in concentration of plasma protein particles at the luminal surface of the BAEC monolayer. It was found that for perfusates containing plasma proteins and/or lipoproteins, water filtration velocity varied as a function of flow rate, being lowest in the absence of flow. Water filtration velocity increased or decreased as flow rate increased or decreased from an arbitrarily set non-zero value, indicating that surface concentration of protein particles varied as a direct function of flow rate, and the process was reversible. It was also found that at particle concentrations equivalent to those found in a culture medium containing serum at 20% by volume, plasma lipoproteins which were much smaller in number and lower in concentration but larger in size than albumin, showed almost the same effect as observed with serum which contained both lipoproteins and albumin, indicating that the substance responsible for this phenomenon is not albumin but lipoprotein whose diffusivity is much smaller than that of albumin. The results strongly support our hypothesis that flow-dependent concentration polarization of lipoproteins occurs at a blood endothelium boundary, and this in turn promote the localization of various vascular diseases which develop in our arterial system. PMID- 10690271 TI - Flow-dependent concentration polarization of plasma proteins at the luminal surface of a semipermeable membrane. AB - The effect of steady shear flow on concentration polarization of plasma proteins and lipoproteins at the luminal surface of a semipermeable vessel wall was studied experimentally using suspensions of these molecules in a cell culture medium and a semipermeable membrane dialysis tube which served as a model of an implanted vascular graft or an artery. The study was carried out by flowing a cell culture medium containing fetal calf serum or bovine plasma lipoproteins or bovine albumin through a 7.5 mm diameter, 60 mm-long dialysis tube in steady flow under a physiologic mean arterial perfusion pressure of 100 mmHg, and measuring the filtration velocity of water (cell culture medium) at the vessel wall which varied as a consequence of the change in concentration of plasma protein particles at the luminal surface of the semipermeable membrane dialysis tube. It was found that for perfusates containing plasma proteins and/or lipoproteins, filtration velocity of water was the lowest in the absence of flow, and it increased or decreased as the flow rate (hence wall shear rate) increased or decreased from a certain non-zero value, indicating that surface concentration of protein particles varied reversibly as a direct function of flow rate. It was also found that at particle concentrations equivalent to those found in a culture medium containing serum at 5% by volume, plasma lipoproteins which were much smaller in number and lower in concentration but larger in size than albumin, had a much larger effect on the filtration velocity of water than albumin. These findings were very much the same as those previously obtained with a cultured endothelial cell monolayer, strongly suggesting that the flow-dependent variation in filtration velocity of water at a vessel wall results from a physical phenomenon, that is, flow-dependent concentration polarization of low density lipoproteins at the luminal surface of the endothelial cell monolayer. PMID- 10690272 TI - Effect of alterations in femoral artery flow on abdominal vessel hemodynamics in swine. AB - In support of an in vivo investigation in swine of the influence of changes in fluid dynamic wall shear on arterial macromolecular permeability, a procedure has been developed to alter the flows in the porcine posterior arterial vasculature by opening and closing a reversible arteriovenous shunt placed on one of the femoral arteries. Laparoscopic techniques were used to place appropriately modified Transonic Systems ultrasonic flow probes on both external and circumflex iliac arteries, and on the terminal aorta. Flow measurements were made prior to shunt placement, and with the shunt open and closed, to measure the influence of altered external iliac artery flow on the distribution to the infrarenal abdominal vessels. Similar experiments were carried out to relate the flow rates in the external iliac arteries to those in the femoral arteries, which are more accessible. Based on the relationships among the measured flow rates, rules have been developed to estimate the major infrarenal flows in the pig, at baseline and with the shunt opened and closed, from only the flow rates measured at the two femoral arteries. PMID- 10690273 TI - Linear viscoelastic model of a maturing gelatin solution. AB - The linear viscoelastic model proposed in this work considers the viscoelastic nature of maturing gelatin solutions through a relaxation modulus that depends on temperature and maturation. This modulus is defined in the conceptual contexts of the classical rubber elasticity theory and the rheometric gel theory. An analysis of the relationship between the equilibrium elastic modulus and the percolation variable around the gel point is also included yielding a percolation exponent close to 1.7 as expected from previous theoretical predictions. Additionally, a simple kinetic model is proposed to follow the microstructural changes obtained as a consequence of the generation of junction zones, the number of which vary with time during the dynamic rheometric tests used in this work. Thus, the storage and loss moduli are measured at different temperatures and frequencies, during the period of gelatin maturation. The theoretical aspects of the rheological model are presented emphasizing the quantitative changes of rheological parameters with the maturation. PMID- 10690274 TI - Wasting away: what a waste: Part 3. PMID- 10690275 TI - Na+/H+ exchange hyperactivity and myocardial hypertrophy: are they linked phenomena? PMID- 10690276 TI - Passive ventricular restraint. PMID- 10690277 TI - The mitochondrial permeability transition and the calcium, oxygen and pH paradoxes: one paradox after another. PMID- 10690278 TI - Oxidative stress, F2-isoprostanes and endothelial dysfunction in hypercholesterolemia. PMID- 10690279 TI - Coming full circle: membrane potential, sarcolemmal calcium influx and excitation contraction coupling in heart muscle. AB - In heart muscle, strong evidence shows that excitation-contraction coupling involves Ca-induced Ca-release. However, under some conditions, single heart cells show Ca release and contraction which is not correlated with Ca entry via the Ca channel, suggesting a second Ca-independent release mechanism. Similar observations were made in early, pioneering studies using voltage-clamped multi cellular preparations. We review the influence that experimental preparations and conditions have had on excitation-contraction coupling theory over the last 20 years. PMID- 10690280 TI - Functional heterogeneity of oxygen supply-consumption ratio in the heart. AB - In this review, the regional heterogeneity of the oxygen supply-consumption ratio within the heart is discussed. This is an important functional parameter because it determines whether regions within the heart are normoxic or dysoxic. Although the heterogeneity of the supply side of oxygen has been primarily described by flow heterogeneity, the diffusional component of oxygen supply should not be ignored, especially at high resolution (tissue regions << 1 g). Such oxygen diffusion does not seem to take place from arterioles or venules within the heart, but seems to occur between capillaries, in contrast to data recently obtained from other tissues. Oxygen diffusion may even become the primary determinant of oxygen supply during obstructed flow conditions. Studies aimed at modelling regional blood flow and oxygen consumption have demonstrated marked regional heterogeneity of oxygen consumption matched by flow heterogeneity Direct, non-invasive indicators of the balance between oxygen supply and consumption include NADH videofluorimetry (mitochondrial energy state) and microvascular PO2 measurement by the Pd-porphyrin phosphorescence technique. These indicators have shown a relatively homogeneous distribution during physiological conditions supporting the notion of regional matching of oxygen supply with oxygen consumption. NADH videofluorimetry, however, has demonstrated large increases in functional heterogeneity of this ratio in compromised hearts (ischemia, hypoxia, hypertrophy and endotoxemia) with specific areas, referred to as microcirculatory weak units, predisposed to showing the first signs of dysoxia. It has been suggested that these weak units show the largest relative reduction in flow (independent of absolute flow levels) during compromising conditions, with dysoxia initially developing at the venous end of the capillary. PMID- 10690281 TI - Gene therapy in the cardiovascular system: an update. AB - This update reviews the remarkable progression in several cardiovascular gene transfer domains. The first chemical gene therapy protocols to stimulate angiogenesis in ischemic myocardium are discussed and both the great expectations as well as remaining hurdle are highlighted. In experimental models of restenosis and heart failure gene therapy shows promising results. Important question regarding vector-related limitations and suboptimal in vivo delivery systems will require expeditious attention for gene therapy to become a more widely applicable option in cardiovascular diseases. PMID- 10690282 TI - Human SCN5A gene mutations alter cardiac sodium channel kinetics and are associated with the Brugada syndrome. AB - BACKGROUND: Primary dysrhythmias other than those associated with the long QT syndrome, are increasingly recognized. One of these are represented by patients with a history of resuscitation from cardiac arrest but without any structural heart disease. These patients exhibit a distinct electrocardiographic (ECG) pattern consisting of a persistent ST-segment elevation in the right precordial leads often but not always accompanied by a right bundle branch block (Brugada syndrome). This syndrome is associated with a high mortality rate and has been shown to display familial occurrence. METHODS AND RESULTS: Pharmacological sodium channel blockade elicits or worsens the electrocardiographic features associated with this syndrome. Hence, a candidate gene approach directed towards SCN5A, the gene encoding the alpha-subunit of the cardiac sodium channel, was followed in six affected individuals. In two patients missense mutations were identified in the coding region of the gene: R1512W in the DIII-DIV cytoplasmic linker and A1924T in the C-terminal cytoplasmic domain. In two other patients mutations were detected near intron/exon junctions. To assess the functional consequences of the R1512W and A1924T mutations, wild-type and mutant sodium channel proteins were expressed in Xenopus oocytes. Both missense mutations affected channel function, most notably a 4-5 mV negative voltage shift of the steady-state activation and inactivation curves in R1512W and a 9 mV negative voltage shift of the steady state activation curve in A1924T, measured at 22 degrees C. Recovery from inactivation was slightly prolonged for R1512W channels. The time dependent kinetics of activation and inactivation at -20 mV were not significantly affected by either mutation. CONCLUSIONS: Two SCN5A mutations associated with the Brugada syndrome, significantly affect cardiac sodium channel characteristics. The alterations seem to be associated with an increase in inward sodium current during the action potential upstroke. PMID- 10690283 TI - Effects of imidapril on NOS expression and myocardial remodelling in failing heart of Dahl salt-sensitive hypertensive rats. AB - OBJECTIVES: To elucidate the relationship between renin-angiotensin system and nitric oxide in hypertensive heart failure, we evaluated the effects of long-term treatment with imidapril, angiotensin-converting enzyme inhibitor, on endothelial cell nitric oxide synthase (eNOS) and inducible NOS (iNOS) expression in the left ventricle (LV) and its relation to myocardial remodelling in failing heart of Dahl salt-sensitive hypertensive rats (DS) fed a high-salt diet. METHODS: In DS rats fed an 8% NaCl diet after the age of 6 weeks, a stage of concentric left ventricular hypertrophy at 11 weeks (DSLVH) was followed by a distinct stage of fatal left ventricular failure with chamber dilatation at 18 weeks (DSCHF). Imidapril (DSCHF-I, n = 7, 1 mg/kg/day, subdepressor dose) or vehicle (DSCHF-V, n = 7) were given from DSLVH to DSCHF stage for 7 weeks, and age-matched (18 weeks) Dahl salt-resistant rats fed the same diet were served as control group (DR-C, n = 7). RESULTS: Markedly increased left ventricular end-diastolic diameter and reduced fractional shortening in DSCHF-V was significantly ameliorated in DSCHF-I using transthoracic echocardiography. The level of eNOS mRNA and protein in the LV was significantly suppressed in DSCHF-V compared with DR-C, and significantly increased in DSCHF-I compared with DR-C and DSCHF-V. The iNOS mRNA and protein and the fibrosis factor expression of type I collagen mRNA were significantly increased in DSCHF-V compared with DR-C, and significantly decreased in DSCHF-I compared with DSCHF-V. DSCHF-V demonstrated a significant increase in wall-to lumen ratio, perivascular fibrosis, and myocardial fibrosis. These changes in the microvasculature were improved significantly by imidapril. CONCLUSIONS: Subdepressor dose of imidapril may ameliorate the endothelial damage not only by inhibiting production of angiotensin II but also by promoting eNOS and inhibiting iNOS mRNA and protein expression in the LV, and this increased eNOS mRNA and protein level may have a role in the improvement of congestive heart failure and myocardial remodelling. PMID- 10690284 TI - Hemodynamic effects of nitric oxide synthase inhibition at steady state and following tumor necrosis factor-alpha-induced myodepression. AB - OBJECTIVE: Nitric oxide (NO) has been proposed as a common mediator of tumor necrosis factor-alpha (TNF alpha)-induced vasodilation and myocardial dysfunction. Accordingly, we performed an extensive assessment of the influence of NO synthase inhibition on left ventricle (LV) and circulatory performance in conscious dogs at steady state and after establishment of TNF alpha mediated myodepression. METHODS: Autonomically blocked, chronically instrumented dogs were studied at steady state and 6 h after initiation of a 1-h rhTNF alpha infusion (40 micrograms/kg). Ventricular performance was evaluated using the pressure volume framework. Dogs were then treated with either NG-nitro-L-arginine methylester (L-NAME, 40 mg/kg bolus) or angiotensin II (250-500 ng/kg). RESULTS: L-NAME, under control conditions or following recombinant human (rh) TNF alpha induced ventricular dysfunction, produced marked increases in afterload with attendant increases in LV pressure, volume, and prolonged isovolumic relaxation without adversely influencing coronary blood flow. regardless of whether the dogs received rhTNF alpha, L-NAME did not affect the slopes of the end-systolic pressure-volume and stroke-work (SW)-end-diastolic volume (EDV) relations (force based measure of contractility), whereas the slope of the dP/dtmax-EDV relation, a velocity dependent parameter of LV systolic function, declined. Overall ventricular performance, as seen by the circulation, was reduced by L-NAME in control as well as rhTNF alpha-treated dogs, evidenced by rightward shifts of the SW-EDV and dP/dtmax-EDV relations. Similar findings were observed in the separate cohorts of dogs, at steady state and 6 h after rhTNF alpha, following angiotensin II at matched systolic pressure. CONCLUSIONS: Systemic NO synthase inhibition with L-NAME does not acutely reverse rhTNF alpha-induced myocardial dysfunction. The detrimental influence of L-NAME on LV size, relaxation, and velocity-based measures of contractility is likely attributable to its effects on increasing afterload. PMID- 10690285 TI - Ischemic preconditioning attenuates ischemia/reperfusion-induced activation of caspases and subsequent cleavage of poly(ADP-ribose) polymerase in rat hearts in vivo. AB - Recently, we have demonstrated that ischemic preconditioning (IP) both limits infarct size and decreases internucleosomal DNA fragmentation in rat hearts in vivo, and that there was a direct correlation between myocardial infarct size and DNA fragmentation even after IP. In this study, we examined the ability of IP to attenuate processing and activation of caspase-1 and caspase-3, and cleavage of poly(ADP-ribose) polymerase (PARP), after prolonged ischemia and reperfusion using the same in vivo animal model. Rats that underwent IP and controls (Ctrl) were subjected to 30 min of left coronary artery occlusion followed by 180 min of reperfusion. IP was accomplished by five 5-min cycles of ischemia, each followed by 5 min of reperfusion. The amount of soluble nucleosomes was measured by enzyme linked immunosorbent assay. Cleavage of caspases-1 and -3, and of one of their substrates PARP, was analyzed by Western blotting. Nucleosomal DNA fragmentation was significantly reduced in ischemic left ventricular (LV) tissue obtained from IP compared with Ctrl animals. The proforms of caspases-1 and -3, and the active form of PARP were not cleaved in the nonischemic LV region of both IP and Ctrl hearts. In contrast, the proform of caspase-3 and the active form of PARP were cleaved in the ischemic LV region of Ctrl hearts, while processing of caspase-1 was increased. Cleavages of caspases-1 and -3, and inactivation of PARP were prevented by IP. The results of this study indicate that IP attenuates both internucleosomal DNA fragmentation and caspases processing, and suggest that the prevention of caspases activation by IP may be important steps in protecting the heart against ischemia/reperfusion injury in vivo. PMID- 10690286 TI - Chronic antisense therapy for angiotensinogen on cardiac hypertrophy in spontaneously hypertensive rats. AB - OBJECTIVE: We examined the effect of the suppression of plasma angiotensinogen (AGT) by the intravenous injection of antisense oligodeoxynucleotides (ODNs) against AGT targeted to the liver on cardiac remodeling in spontaneously hypertensive rats (SHR). The ODNs against rat AGT were coupled to asialoglycoprotein (ASOR) carrier molecules, which serve as an important method for regulating liver gene expression. METHODS: Male SHR (n = 18), and age-matched male Wistar-Kyoto rats (WKY, n = 6) were used for this study. At 10 weeks of age, the SHR were divided into three groups (each group n = 6), and the systolic blood pressure (SBP) did not significantly change among them. The control SHR and WKY groups received saline, the sense SHR group was injected with the sense ODNs complex and the antisense SHR group was injected with the antisense ODNs complex, from 10 to 20 weeks of age. ASOR-poly(L)lysine-ODNs complex was injected via the tail veins twice a week. RESULTS: At the end of the treatment, a reduction of hepatic AGT mRNA, cardiac angiotensin II type 1 receptor mRNA and the plasma AGT concentration was only observed in the antisense-injected SHR but not in the other groups of SHR and WKY. This antisense therapy did not significantly change the mRNA expression for angiotensin converting enzyme, angiotensin II type 2 receptor and AGT in the left ventricle (LV) among all three groups. Although the plasma angiotensin II (Ang II) concentration significantly decreased to the level of WKY after the antisense therapy, the SBP, LV to body weight ratio and % collagen volume fraction also showed a reduction, however, these findings were still larger than in the WKY than in either the sense-injected SHR or control SHR. CONCLUSION: The plasma AGT is considered to play a role in the development of cardiac hypertrophy in SHR, but it has not a complete effects on cardiac remodeling even if the plasma Ang II levels are inhibited because of an insufficient suppression of hypertension. PMID- 10690287 TI - Passive ventricular constraint amends the course of heart failure: a study in an ovine model of dilated cardiomyopathy. AB - OBJECTIVE: Dilated cardiomyopathy (DCM) is associated with a progressive deterioration in cardiac function. We hypothesised that some of the deleterious effects of DCM could be reduced by mechanically limiting the degree of cardiac dilatation. METHODS: A Transonic 20A cardiac output (CO) flow-probe was implanted in the pulmonary artery of 12 adult (52 +/- 4 kg) sheep. Early heart failure was created by rapid right ventricular (RV) pacing for 21 days at a rate which resulted in an initial 10% decrease in CO (to a maximum of 190 bpm). A custom polyester jacket (Acorn Cardiovascular, St Paul, MN) was then placed, via a partial lower sternotomy, on the ventricular epicardium of all sheep. Animals were randomised either to jacket retention (wrap) or removal (sham). Pacing was recommenced at a higher rate (that initiated a further 10% decrease in CO) for 28 days. Haemodynamic and echocardiographic parameters were determined at baseline, implant and at termination. RESULTS: At termination, the left ventricular fractional shortening was significantly higher (p = 0.03), the degree of mitral valve regurgitation lower (scaled 0-3) (p = 0.03) and the left ventricular long axis area smaller (p = 0.02) in the wrap animals compared with sham. CONCLUSIONS: In this model of heart failure, ventricular constraint with a polyester jacket diminished the deterioration in cardiac function associated with progressive dilated cardiomyopathy. These results suggest that maintainance of a more normal cardiac size and shape may be beneficial in patients with dilated cardiomyopathy. PMID- 10690288 TI - On the mechanism of the failure of mitochondrial function in isolated guinea-pig myocytes subjected to a Ca2+ overload. AB - OBJECTIVE: The influence of agents that inhibit the movement of Ca2+ across the mitochondrial membrane or Ca2+ dependent changes to this membrane upon the response of isolated ventricular myocytes to a Ca2+ overload has been investigated. METHODS: The changes of intracellular Ca2+ and Mg2+ ([Ca2+]i, [Mg2+]i) (as reflected by cellular ATP), mitochondrial membrane potential (psi m) and NADH was measured upon the response of isolated ventricular myocytes to a Ca2+ overload. RESULTS: A slow depolarization of psi m during Ca2+ depletion and its prompt recovery on Ca2+ repletion were unaffected by ruthenium red, clonazepam, CGP-37157 which is a high potent inhibitor of the mitochondrial Na+/Ca2+ antiport or cyclosporin A but a large delayed sustained depolarization was inhibited. The slow small fall in [Mg2+]i on Ca2+ depletion and a rapid recovery on Ca2+ repletion were unaffected by ruthenium red, clonazepam, CGP 37157 or cyclosporin A. A delayed sustained larger rise in [Mg2+]i was inhibited. The marked sustained fall in NADH autofluorescence that occurs on Ca2+ overload was attenuated and transient in the presence of ruthenium red, CGP-37157 and cyclosporin A. CONCLUSION: These results are consistent with an increase in Ca2+ cycling across the mitochondrial membrane provoked by the combined Na+ and Ca2+ overload of cardiac myocytes, causing a depolarization sufficient to uncouple respiration and lead to the depletion of cellular ATP. PMID- 10690289 TI - Norpropoxyphene-induced cardiotoxicity is associated with changes in ion selectivity and gating of HERG currents. AB - OBJECTIVE: Norpropoxyphene (NP) is a major metabolite of propoxyphene (P), a relatively weak mu-opioid receptor agonist. Toxic blood concentrations ranging from 3 to 180 mumol/l have been reported and the accumulation of NP in cardiac tissue leads to naloxone-insensitive cardiotoxicity. Since several lines of evidence suggest that not only block of INa but also IK block may contribute to the non-opioid cardiotoxic effects of P and NP, we investigated the effects of P and NP on HERG channels. HERG presumably encodes IKr, the rapidly-activating delayed rectifier K+ current, which is known to have an important role in initiating repolarization of action potentials in cardiac myocytes. METHODS: Using the 2-microelectrode voltage clamp technique we investigated the interaction of P and NP with HERG channels, expressed in Xenopus oocytes. RESULTS: Our experiments show that low drug concentrations (5 mumol/l) facilitate HERG currents, while higher drug concentrations block HERG currents (IC50-values of approx. 40 mumol/l) and dramatically shift the reversal potential to a more positive value because of a 30-fold increased Na(+)-permeability. P and NP also alter gating of HERG channels by slowing down channel activation and accelerating channel deactivation kinetics. The mutant S631C nullifies the effect of P and NP on the channel's K(+)-selectivity. CONCLUSION: P and NP show a complex and unique drug-channel interaction, which includes altering ion-selectivity and gating. Site-directed mutagenesis suggests that an interaction with S631 contributes to the drug-induced disruption of K(+)-selectivity. No specific role of the minK subunit in the HERG block mechanism could be determined. PMID- 10690290 TI - The angiotensin-converting enzyme inhibitor, fosinopril, and the angiotensin II receptor antagonist, losartan, inhibit LDL oxidation and attenuate atherosclerosis independent of lowering blood pressure in apolipoprotein E deficient mice. AB - OBJECTIVE: To investigate the possible mechanisms of the antiatherosclerotic effects of the angiotensin-converting enzyme (ACE) inhibitor, fosinopril, in apolipoprotein (apo) E deficient mice. METHODS: Apo E deficient (E0) mice at the age of 8 weeks received either placebo or a high dose (25 mg/kg/d) of fosinopril supplemented in their drinking water. RESULTS: After 12 weeks of treatment, fosinopril reduced the aortic lesion size by 70%, compared with the placebo group. At this dosage, fosinopril significantly reduced blood pressure from 93 +/ 2 mmHg before treatment to 70 +/- 2 mmHg at the end of the treatment period (P < 0.005). Fosinopril also increased the resistance of the mice plasma low density lipoprotein (LDL) to CuSO4-induced oxidation, as shown by a 90% reduction in the LDL content of malondialdehyde (MDA) and also by a prolongation of the lag time required for the initiation of LDL oxidation (from 100 min in the placebo-treated mice to more than 240 min in the fosinopril-treated mice; P < 0.001). In addition, fosinopril inhibited CuSO4-induced oxidation of LDL that was obtained from the aortas of the treated mice, as shown by an 18% and 37% reduction in the LDL content of lipid peroxides and hydroperoxy-cholesterol linoleate, respectively, compared with the placebo-treated mice (P < 0.01). A low dosage of fosinopril (5 mg/kg/d) that was still adequate to reduce their plasma ACE activity and LDL propensity to lipid peroxidation was insufficient to lower their blood pressure. This dosage also reduced the aortic lesion size in the apo E deficient mice by 40% (P < 0.01). CONCLUSIONS: The antiatherogenic effects of fosinopril in apo E deficient mice are due not only to blood pressure reduction but also to the direct inhibition of angiotensin II-dependent effects, which are probably also associated with the inhibition of LDL oxidation. PMID- 10690291 TI - Peroxisome proliferator-activated receptor gamma C161-->T polymorphism and coronary artery disease. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPAR gamma) as a transcription factor plays an important role in lipid metabolism, glucose homeostasis, insulin sensitivity, obesity, diabetes, foam cell formation and atherogenesis. METHODS AND RESULTS: We have studied distribution of the PPAR gamma C161-->T substitution at exon 6 in 647 Australian Caucasian patients aged < or = 65 years (484 men and 163 women) recruited consecutively, with or without angiographically documented coronary artery disease (CAD). The frequencies of the CC, CT and TT genotypes were 69.8%, 27.7% and 2.5% and the 'T' allele frequency 0.163. They were in Hardy-Weinberg equilibrium and not different between men and women. The BMI and waist to hip ratio (WHR) among patients with CC, CT + TT genotypes were not different (P = 0.878, P = 0.677). However there was a significant association between the polymorphism and CAD. The 'T' allele carriers (CT + TT) had significantly reduced CAD risk compared to the CC homozygotes (odds ratio: 0.457, 95% CI: 0.273-0.763, P = 0.0045) in a logistic regression model after controlling other known risk factors. This reduced risk was particularly evident among CT heterozygotes (odds ratio: 0.466, 95% CI: 0.291-0.746, P = 0.0015), who also had lower apo B and total cholesterol to HDL-C ratios (P < 0.05). CONCLUSION: We report that the PPAR gamma C161-->T substitution is associated with a reduced CAD risk, particularly among CT heterozygous patients, but not with obesity in Australian Caucasian patients. It implicates that the PPAR gamma may have a significant role in atherogenesis, independent of obesity and of lipid abnormalities, possibly via a direct local vascular wall effect. PMID- 10690292 TI - Effects of brain natriuretic peptide on forearm vasculature: comparison with atrial natriuretic peptide. AB - OBJECTIVE: The aim of the present study was to determine the vasoactive effects of brain natriuretic peptide (BNP) as compared to those of atrial natriuretic peptide (ANP) in normal man. METHODS: Ten healthy male subjects (median age 21 (20-23) year) were studied twice. In the first study equimolar doses (1, 3, and 10 pmol/dl/min) of both BNP and ANP (in random order and double blind) were infused into the brachial artery of the non-dominant arm with a 1-h wash-out period in between. In the second study two BNP (n = 5) or ANP (n = 5) dose response curves were performed in order to assess the repeatability of the BNP/ANP infusions. To this end, BNP and ANP were infused in the same equimolar doses as in the first protocol. Forearm blood flow (FBF) was determined by venous occlusion plethysmography before and during infusions. RESULTS: BNP increased the FBF ratio (infused/contralateral arm) by 6%, 17%, and 48%, respectively (p < 0.05), while ANP increased the FBF ratio by 4%, 58%, and 133% (p < 0.001). The slopes of the BNP dose-response curves differed significantly from those of the ANP curves (18.1 versus 43.2; p = 0.022). No differences were observed between the repeated dose-response curves of either BNP or ANP. CONCLUSIONS: The present data demonstrate that BNP induces a dose-dependent vasodilatation in man. On a molar basis, however, this vasodilatation is significantly less than the vasodilatation induced by ANP. These differences may be related to differences in natriuretic-peptide-receptor affinity. Furthermore, our data show that the vasoactive effects of both BNP and ANP are repeatable in time. PMID- 10690293 TI - Enhanced coronary vasoconstriction to oxidative stress product, 8-epi prostaglandinF2 alpha, in experimental hypercholesterolemia. AB - OBJECTIVES: The F2-isoprostanes are a family of novel prostaglandin isomers and a stable product of in vivo oxidative stress. 8-epi-prostaglandinF2 alpha, a member of this isoprostane family, is a vasoconstrictor and its local release may contribute to the abnormal vasomotor tone associated with hypercholesterolemia. We therefore aimed to outline the role of 8-epi-prostaglandinF2 alpha as a coronary vasoconstrictor in experimental hypercholesterolemia. METHODS AND RESULTS: Pigs were randomized to two experimental groups (each n = 9): normal (N) and high cholesterol (HC) diet. To determine the vasoconstrictive effects of 8 epi-prostaglandinF2 alpha in vitro, doses from 10(-9) to 10(-5) M were used to constrict coronary epicardial rings. Plasma total and LDL cholesterol levels were significantly higher in the HC group compared with the N group (P < 0.005) as were plasma 8-epi-prostaglandinF2 alpha levels (P < 0.001). 8-epi-prostaglandinF2 alpha immunoreactivity was present in the vessel wall in both groups. Normal vessels with intact endothelium (n = 8 rings) contracted to 8-epi-prostaglandinF2 alpha (maximal contraction 15.5 +/- 8.74%). In the HC group, rings with intact endothelium had a greater contractile response to 8-epi-prostaglandinF2 alpha compared to normals (72.3 +/- 7.9%; n = 8; P < 0.0001). This was reversed by preincubation with NOR-3, a NO donor (maximal contraction 6.7 +/- 1.56%; n = 5; P < 0.0001). Enhanced contraction in normal vessels occurred with endothelial denudation (98.4 +/- 3.56%; n = 6; P < 0.0001) and with preincubation of the endothelium-intact rings with L-NMMA (N-monomethyl-L-arginine), an NO synthase inhibitor (85.5 +/- 10.3%, n = 6, P < 0.001). The enhanced contraction seen with hypercholesterolemia did not occur with other prostanoid vasoconstrictors. CONCLUSION: Experimental hypercholesterolemia leads to a significant increase in 8-epi-prostaglandinF2 alpha levels in addition to enhanced 8-epi-prostaglandinF2 alpha-induced coronary vasoconstriction, in vitro. These findings support a role for the F2-isoprostanes in the regulation of coronary vasomotor tone in pathophysiologic states. PMID- 10690294 TI - Estrogen attenuates the adventitial contribution to neointima formation in injured rat carotid arteries. AB - OBJECTIVE: This study tested, in ovariectomized rats, whether (1) adventitial activation plays a role in the vascular injury response, and (2) inhibition of adventitial activation and the subsequent wave of cell proliferation moving from adventitia to neointima contributes to the estrogen-induced attenuation of neointima formation in balloon injured carotid arteries. METHODS: Ovariectomized Sprague-Dawley rats were treated with either 17 beta-estradiol or vehicle beginning 72 h prior to balloon injury of the right common carotid artery and were sacrificed at 0, 3, 7, 14 and 28 days after injury. BrdU was administered 18 h and 12 h prior to sacrifice in order to quantitate mitotic activity in adventitia, media and neointima of the damaged vessel at specified times post injury. RESULTS: Adventitial activation, evidenced by positive BrdU staining, was evident on the day of injury, peaked on day 3 and was resolved by day 7, thus preceding neointima formation. Numbers of BrdU labeled cells in adventitia on day 3 were significantly reduced in estrogen treated rats compared to controls. BrdU labeled cells were undetectable in media on the day of injury, appeared at day 3 and disappeared by day 14. Neointima appeared at day 7 and increased in area throughout the period of observation. Neointimal area and numbers of BrdU labeled cells in neointima were significantly reduced in estrogen treated rats compared to controls. These findings suggest that there is a wave of cell proliferation moving in an adventitia-to-lumen direction following endoluminal injury of the rat carotid artery and that estrogen modulates this proliferative response to injury. CONCLUSION: These results support the hypothesis that adventitial activation contributes to the vascular injury response and that estrogen reduces this contribution. PMID- 10690295 TI - Chronic cardiac denervation affects the speed of coronary vascular regulation. AB - OBJECTIVE: We tested the hypothesis that the rate of adaptation of coronary metabolic vasodilatation and autoregulation is modulated by the cardiac nerves. METHODS: Anaesthetised dogs (seven innervated (control) and seven with denervated hearts) were subjected to controlled pressure perfusion of the left main coronary artery. Heart rate was controlled by pacing. RESULTS: The steady state autoregulation curves and metabolic regulation curves were similar in the two groups. A sudden increase or decrease in heart rate was associated with a faster response (22% shorter half-times) in the innervated than the denervated dogs (P < 0.001). A sudden increase or decrease in coronary arterial perfusion pressure was associated with a slower response (24% longer half-times) in the innervated than the denervated hearts (P < 0.005). CONCLUSIONS: We conclude that the speed of response to metabolic and perfusion pressure changes is partly mediated by cardio cardiac reflexes. Reflex coronary vasodilatation appears to reinforce the metabolic vasodilatation of a heart rate increase and oppose the vasoconstriction in response to increased perfusion pressure. PMID- 10690296 TI - Involvement of myosin light-chain kinase in chloride-sensitive Ca2+ influx in porcine aortic endothelial cells. AB - OBJECTIVES: This study was designed to investigate the involvement of myosin light-chain kinase (MLCK) in bradykinin- and thapsigargin-induced changes in intracellular Cl- and Ca2+ concentrations ([Cl-]i; [Ca2+]i) in porcine aortic endothelial cells. METHODS: Using the fluorescent probes N-ethoxycarbonylmethyl-6 methoxyquinolinium bromide (MQAE) and fura-2/AM, the effects of different MLCK inhibitors on bradykinin- and thapsigargin-induced changes in [Cl-]i and [Ca2+]i were assessed. RESULTS: Bradykinin and thapsigargin significantly decreased the MQAE fluorescence intensity, which indicates increased [Cl-]i; these changes were reversed by removal of extracellular chloride (Cl-o) and were significantly inhibited by Cl(-)-channel inhibitor N-phenylanthranilic acid but not by Na(+) K(+)-Cl- cotransport inhibitor furosemide. Pretreatment with ML-9 and wortmannin, two different selective inhibitors of MLCK, significantly reduced these changes in a dose-dependent manner. The inhibitory effects of ML-9 and wortmannin on the Cl- responses were not significantly different and were not additive. Bradykinin and thapsigargin provoked large increases in [Ca2+]i, which were significantly diminished by removal of Cl-o and by pretreatment with the Cl(-)-channel inhibitor N-phenylanthranilic acid. CONCLUSIONS: The study shows that an increase in [Cl-]i may be involved in the Ca2+ influx in response to bradykinin and thapsigargin and that MLCK might be involved in the Cl- response. We suggest that MLCK might be involved in the Cl(-)-sensitive endothelial Ca2+ responses to bradykinin and thapsigargin. PMID- 10690297 TI - Wasting away. What a waste. Part 2. PMID- 10690298 TI - The infarcted myocardium: simply dead tissue, or a lively target for therapeutic interventions. PMID- 10690299 TI - A plethora of mechanisms in the HERG-related long QT syndrome. Genetics meets electrophysiology. PMID- 10690300 TI - Rapid estimation of myocardial infarct size. PMID- 10690301 TI - Rate control in atrial fibrillation: role of atrial inputs to the AV node. PMID- 10690302 TI - Is there a local renin-angiotensin system in the heart? AB - The existence of a local renin-angiotensin system in the heart is still a controversial issue. This review discusses the evidence, obtained from studies in cardiac cells, in isolated perfused hearts and in intact animals and humans, both under normal and pathological conditions, for local production of prorenin, renin, angiotensinogen, angiotensin-converting enzyme, angiotensin I and angiotensin II at cardiac tissue sites. In addition, the role of alternative angiotensin-generating enzymes (cathepsin, chymase) and the possibility of (pro)renin uptake from the circulation is evaluated. PMID- 10690303 TI - Actin-myosin interaction. AB - Recent advances in the study of muscle physiology was made possible by the application of novel experimental techniques including in vitro motility assay, molecular biology, and X-ray crystallography. A similar approach was successfully applied in studying the properties of cardiac actin-myosin interaction. Implication in clinical cardiology is also reviewed. PMID- 10690304 TI - Role of endothelial and smooth muscle cells in the physiopathology and treatment management of pulmonary hypertension. AB - Pulmonary hypertension can occur either as primary or secondary disease following cardiac or pulmonary illnesses. In either cases, histological lung biopsies reveal vascular remodelling i.e. smooth muscle cells proliferation with medial hypertrophy, arteriolar muscularization and endothelial cell proliferation. Subsequent intimal thickening, fibrosis and in situ thrombosis, altogether lead to vaso-occlusive alterations referred to as plexiform lesions. Theories concerning the detailed physiopathology of pulmonary hypertension have focused on endothelial and smooth muscle cells' chemical factors production and response to different mediators. The endothelium produces vasoconstrictor and growth promoting factor such as endothelin-1 (ET-1) as well as vasodilator and growth inhibitor factors like prostacyclin and nitric oxide (NO). ET-1 has been noted in high concentrations in some clinical cases and experimental models of pulmonary hypertension, coupled with ET-1 receptors' modulation and altered endothelin converting enzyme activities, suggesting their active role in both arteriolar vasoconstriction and occlusion. Vascular thrombosis which has been noted by pathologists in pulmonary hypertension, could be related to an imbalance between thrombotic inducing factors (such as anti-phospholipid antibodies, ET-1 and thromboxane) and decreased fibrinolytic activity and antiaggregant endothelial factors (like prostacyclin, NO, thrombomodulin). The discovery of an endothelial cells' monoclonal proliferation in plexiform lesions of patients with primary pulmonary hypertension may reinforce the cellular proliferation hypothesis to understand the histopathology of this disease. In view of these new findings, the treatments available for pulmonary hypertension have been expanded from the previously employed oxygen therapy, calcium-channel blockers and anticoagulants, to intravenous prostacyclin analogues (epoprostenol) and inhaled nitric oxide. PMID- 10690305 TI - Voltage-shift of the current activation in HERG S4 mutation (R534C) in LQT2. AB - OBJECTIVE: Recently, a novel missense mutation (R534C) in the S4 region of human ether-a-go-go-related gene (HERG) was identified in one Japanese LQT2 family. The S4 region presumably functions as a voltage sensor. However, it has not yet been addressed whether the S4 region of HERG indeed functions as a voltage sensor, and whether these residues play any role in abnormal channel function in cardiac repolarization. METHODS: We characterized the electrophysiological properties of the R534C mutation using the heterologous expression system in Xenopus oocytes. Whole cell currents were recorded in oocytes injected with wild-type cRNA, R534C cRNA, or a combination of both. RESULTS: Clinical features--QTc intervals of all affected patients with R534C mutation in HERG are prolonged ranging from 460 to 680 ms (averaged QTc interval > 540 ms). One member of this family had experienced sudden cardiac arrest, and other suffered from recurrent palpitation. Electrophysiology--Oocytes injected with R534C cRNA did express functional channels with altered channel gating. Kinetic analyses revealed that the R534C mutation shifted the voltage-dependence of HERG channel activation to a negative direction, accelerated activation and deactivation time course, and reduced steady-state inactivation. Quantitative analyses revealed that this mutation did not cause apparent dominant-negative suppression. Computer simulation- Incorporating the kinetic alterations of R534C, however, did not reproduce prolonged action potential duration (APD). CONCLUSIONS: The data revealed that arginine at position 534 in the S4 region of HERG is indeed involved in voltage dependence of channel activation as a voltage sensor. Our examination indicated that HERG current suppression in R534C mutation was the least severe among other mutations that have been electrophysiologically examined, while affected patients did show significant QT prolongation. This suggest that another unidentified factor(s) that prolong APD might be present. PMID- 10690306 TI - Adenoviral-mediated gene transfer induces sustained pericardial VEGF expression in dogs: effect on myocardial angiogenesis. AB - OBJECTIVE: Angiogenic peptides like VEGF (vascular endothelial growth factor) and bFGF (basic fibroblast growth factor) have entered clinical trials for coronary artery disease. Attempts are being made to devise clinically relevant means of delivery and to effect site-specific delivery of these peptides to the cardiac tissue, in order to limit systemic side-effects. We characterized the response of the pericardium to delivery of a replication-deficient adenovirus carrying the cDNA for AdCMV.VEGF165, and assessed the effect of pericardial VEGF165 on myocardial collateral development in a canine model of progressive coronary occlusion. METHODS: Ameroid constrictors were placed on the proximal left circumflex coronary artery of mongrel dogs. Ten days later, 6 x 10(9) pfu AdCMV.VEGF165 (n = 9). AdRSV.beta-gal (n = 9), or saline (n = 7) were injected through an indwelling pericardial catheter. Transfection efficiency was assessed by X-gal staining. Pericardial and serum VEGF levels were measured serially by ELISA. Maximal myocardial collateral perfusion was quantified with radiolabeled or fluorescent microspheres 28 days after treatment. RESULTS: In AdRSV.beta-gal treated dogs, there was extensive beta-gal staining in the pericardium and epicardium, with minimal beta-gal staining in the mid-myocardium and endocardium. Pericardial delivery of AdCMV.VEGF165 resulted in sustained (8-14 day) pericardial transgene expression, with VEGF levels peaking 3 days after infection (> 200 ng/ml) and decreasing thereafter. There was no detectable increase in serum VEGF levels. Maximal collateral perfusion, a principal correlate of collateral development and angiogenesis, was equivalent in all groups. CONCLUSION: Adenoviral-mediated gene transfer is capable of inducing sustained VEGF165 expression in the pericardium; however, locally targeted pericardial VEGF delivery failed to improve myocardial collateral perfusion in this model. PMID- 10690307 TI - Effects of chronic treatment by amiodarone on transmural heterogeneity of canine ventricular repolarization in vivo: interactions with acute sotalol. AB - OBJECTIVE: The present study was designed to examine the effects of chronic amiodarone on the different ventricular cell subtypes in situ and to evaluate its interactions with sotalol. METHODS: Three groups of dogs were studied. Group I (n = 8) received no treatment. Group II (n = 7) and group III (n = 8) received, respectively, 100 and 200 mg amiodarone orally twice a day for 6 weeks to 8 months. In vivo studies were performed under halothane anesthesia 14 h after the last administration of amiodarone. Three leads ECG, femoral blood pressure and left ventricular intramural monophasic action potentials (MAP) were continuously recorded. Bradycardia was obtained by clamping the sinus node and beta-blockade and the heart was driven by atrial pacing. Three weeks before the in vivo experiments, the cellular electrophysiologic properties of right ventricular tissues obtained by cardiac biopsy in six treated and six control dogs were studied with standard microelectrodes. RESULTS: Amiodarone produced a dose dependent decrease in plasma levels of triiodothyronine (T3; 5.9 +/- 0.4 pM in control dogs, 3.1 +/- 0.2 pM in group III, P < 0.001) without affecting thyroxine (T4). Under anesthesia, the QT interval was 14% larger in group III compared to group I at a paced cycle length (PCL) of 1500 ms (P < 0.05). This is consistent with the 10% increase in endocardial MAP duration in group III at the same PCL (P < 0.05). There was no significant increase in transmural dispersion of MAP duration. In group I, sotalol induced a significant reverse use-dependent increase in MAP duration. This effect was reduced in group II and completely suppressed in group III. Amiodarone prevented the sotalol-induced increase in transmural dispersion of ventricular repolarization which was 69 +/- 12 ms in untreated dogs, 41 +/- 8 ms in group II (P < 0.05) and 34 +/- 8 ms (P < 0.05) in group III at PCL = 1500 ms. Amiodarone also prevented the sotalol-induced ventricular tachyarrhythmias. In vitro, the action potential duration was longer in amiodarone-treated dogs that in control ones (208 +/- 5 ms versus 188 +/- 9 ms at PCL = 1000 ms, P < 0.05). The sotalol-induced prolongation of repolarization was reduced in amiodarone-treated dogs. CONCLUSION: Chronic treatment of dogs with amiodarone induced a moderate prolongation of the QT interval and MAP duration without affecting transmural dispersion of repolarization and inhibited the effects of acute sotalol, including the prolongation of repolarization, the increase in transmural dispersion of repolarization and the induction of arrhythmias. PMID- 10690308 TI - Measurement of myocardial infarct size from plasma fatty acid-binding protein or myoglobin, using individually estimated clearance rates. AB - OBJECTIVE: In patients with acute myocardial infarction (AMI), estimation of infarct size from the early markers, fatty acid-binding protein (FABP) and myoglobin (MYO), usually assumes average (fixed) rate constants (FCR) for protein clearance from plasma. However, individual variation in FCR is large. Renal dysfunction causes slower clearance of FABP and MYO from plasma and, hence, overestimation of infarct size in 20-25% of patients. We investigated whether or not more accurate values of infarct size could be obtained with individually estimated clearance rates. METHODS: Concentrations of FABP and MYO and, for comparison, activities of the established cardiac markers, creatine kinase (CK) and alpha-hydroxybutyrate dehydrogenase (HBDH), were assayed in serial plasma samples from 138 patients with AMI. Individual FCR values of FABP and MYO were estimated from plasma creatinine concentrations, sex and age. RESULTS: Individual FCR values varied from 0.4 to 2.4 h-1. Use of these individual FCR values significantly improved the correlation between infarct size, as estimated from FABP or MYO on the one hand, and from CK and HBDH on the other. Approximately equal estimates of infarct size were obtained for all four marker proteins. CONCLUSIONS: Using individually estimated clearance rates, renal insufficiency no longer hampers calculation of infarct size from FABP and MYO, and reliable estimates of total myocardial damage can be obtained within 24 h after first symptoms. PMID- 10690309 TI - Nitric oxide controls cardiac substrate utilization in the conscious dog. AB - OBJECTIVES: The aim of this study was to determine whether the acute inhibition of nitric oxide (NO) synthase causes changes in cardiac substrate utilization which can be reversed by a NO donor. METHODS: NO synthase was blocked by giving 30 mg/kg of nitro-L-arginine (NLA) i.v. to 15 chronically instrumented dogs. Hemodynamics and blood samples from aorta and coronary sinus were taken at control and at 1 and 2 h after NLA. In five dogs, 0.4 mg/kg of the NO donor 3754 was given i.v. 1 h after NLA. In six dogs, angiotensin II was infused over 2 h (20-40 ng/kg/min) to mimic the hemodynamic effects of NLA. RESULTS: Two h after NLA: mean arterial pressure was 153 +/- 4 mmHg; MVO2 increased by 38%; cardiac uptake of lactate and glucose increased, respectively, from 20.0 +/- 5.0 to 41.0 +/- 9.3 mumol/min and from 1.1 +/- 0.7 to 6.8 +/- 1.5 mg/min (all P < 0.05 vs. control). Cardiac uptake of free fatty acids decreased by 43% after 1 h (P < 0.05) and returned to control values at 2 h. Cardiac respiratory quotient increased from 0.76 +/- 0.03 to 1.05 +/- 0.07, indicating a shift to carbohydrate oxidation. All these changes were reversed by the NO donor. In the dogs receiving angiotensin II infusion, MVO2 increased by 28% and lactate uptake doubled (both P < 0.05), but no other metabolic changes where observed. CONCLUSIONS: The acute inhibition of NO synthase by NLA causes a switch from fatty acids to lactate and glucose utilization by the heart which can be reversed by a NO donor, suggesting an important regulatory action of NO on cardiac metabolism. PMID- 10690310 TI - Increased hypoxic stress decreases AMP hydrolysis in rabbit heart. AB - OBJECTIVE: AMP conversion to adenosine by cytosolic 5'nucleotidase (5NT) or to IMP by AMP deaminase determines the degree of nucleotide degradation, and thus ATP resynthesis, during reoxygenation. To elucidate the regulation of AMP hydrolysis during ischemia, data from 31P NMR spectroscopy and biochemical analyses were integrated via a mathematical model. Since 5NT is downregulated during severe underperfusion (5% flow), we tested 5NT regulation during less severe underperfusion (10% flow) and then made the perfusate hypoxic to see if the greater stress reactivated 5NT. METHODS: 31P NMR spectra and coronary venous effluents were obtained from Langendorff-perfused rabbit hearts subjected to two 30-min periods of underperfusion (10% flow); the second period with or without additional hypoxia (30% O2). Data were analyzed with a mathematical model describing the kinetics of myocardial energetics and metabolism. RESULTS: A single 30-min period of 10% flow causes downregulation of AMP hydrolysis and the data from the second period of underperfusion are best described by lower 5NT activity, even in the presence of extra hypoxia. Thirty percent less purines appear in the venous effluent than predicted by the phosphoenergetics (PCr and ATP) when IMP is not allowed to accumulate by the model, however the model indicates that a constant accumulation of IMP via AMP deaminase could explain the discrepancy between expected and measured purines in the venous effluent. CONCLUSIONS: While AMP hydrolysis to adenosine is prominent in early ischemia and acts to preserve cellular energy potential, during a second ischemic period, nucleotides are conserved by the stable inhibition of AMP hydrolysis. Furthermore, during 10% flow conditions, nucleotides are conserved, possibly via an IMP-accumulatory pathway. PMID- 10690311 TI - Role of the differential bombardment of atrial inputs to the atrioventricular node as a factor influencing ventricular rate during high atrial rate. AB - OBJECTIVES: The role of the atrial inputs for the conduction through the atrioventricular node (AVN) at slow rates and during reentrant tachycardia is well acknowledged, although still controversial. However, the relationship between the sequence and rate of atrial engagement of the AVN inputs and the resulting ventricular rate during high atrial rate remains unclear. This study provides quantitative description of complex AVN input-output correlations determining the ventricular rate during random high atrial rate. METHODS AND RESULTS: 12 rabbit heart preparations were used to evaluate the ventricular rate during programmed regular high atrial rate pacing or random pacing from eight atrial sites. Electrograms were recorded at the posterior (P) and anterior (A) AVN inputs, and at the bundle of His along with nodal cellular action potentials. Lorenz-plots and input-output-rate correlations were used to quantify the ventricular rate under different pacing protocols. Small alternations in the sequence of activation of P and A resulted in substantial changes of the organization of the intranodal cellular responses and the ventricular rate. The ventricular rate was shown to be significantly dependent on the site of high rate pacing (P < 0.01) and on the resulting mean rate of inputs activation. Furthermore, the asymmetry between P- and A-bombardment was an important determinant, so that high ventricular rate was associated with large difference between the inputs rates and vice versa (P < 0.05). CONCLUSIONS: The prevailing ventricular rate during high atrial rate is a complex dynamic parameter that depends not only on the global mean atrial rate but, in a major part, on the differential bombardment of the AVN inputs and on the site of initiation of the atrial wave fronts. PMID- 10690312 TI - Identification and properties of ATP-sensitive potassium channels in myocytes from rabbit Purkinje fibres. AB - OBJECTIVE: Our goal was to identify the ATP-sensitive potassium (KATP) channels in cardiac Purkinje cells and to document the functional properties that might distinguish them from KATP channels in other parts of the heart. METHODS: Single Purkinje cells and ventricular myocytes were isolated from rabbit heart. Standard patch-clamp techniques were used to record action potential waveforms. and whole cell and single-channel currents. RESULTS: The KATP channel opener levcromakalim (10 microM) caused marked shortening of the Purkinje cell action potential. Under whole-cell voltage-clamp, levcromakalim induced an outward current, which was blocked by glibenclamide (5 microM), in both Purkinje cells and ventricular myocytes. Metabolic poisoning of Purkinje cells with NaCN and 2-deoxyglucose caused a significant shortening of the action potential (control 376 +/- 51 ms; 6 min NaCN/2-deoxyglucose 153 +/- 21 ms). This effect was reversed with the application of glibenclamide. Inside-out membrane patches from Purkinje cells showed unitary current fluctuations which were inhibited by cytoplasmic ATP with an IC50 of 119 microM and a Hill coefficient of 2.1. This reflects approximately five-fold lower sensitivity to ATP inhibition than for KATP channels from ventricular myocytes under the same conditions. The slope conductance of Purkinje cell KATP channels, with symmetric, 140 mM K+, was 60.1 +/- 2.0 pS (mean +/- SEM). Single-channel fluctuations showed mean open and closed times of 3.6 +/- 1.5 ms and 0.41 +/- 0.2 ms, respectively, at -60 mV and approximately 21 degrees C. At positive potentials. KATP channels exhibited weak inward rectification that was dependent on the concentration of internal Mg2+. Computer simulations, based on the above results, predict significant shortening of the Purkinje cell action potential via activation of KATP channels in the range 1-5 mM cytoplasmic ATP. CONCLUSIONS: Purkinje cell KATP channels may represent a molecular isoform distinct from that present in ventricular myocytes. The presence of KATP channels in the Purkinje network suggests that they may have an important influence on cardiac rhythm and conduction during periods of ischemia. PMID- 10690313 TI - Connexin expression in cultured neonatal rat myocytes reflects the pattern of the intact ventricle. AB - OBJECTIVE: Primary cultures of neonatal rat ventricular myocytes have become a widely used model to examine a variety of functional, physiological and biochemical cardiac properties. In the adult rat, connexin43 (Cx43) is the major gap junction protein present in the working myocardium. In situ hybridization studies on developing rats, however, showed that Cx40 mRNA displays a dynamic and heterogeneous pattern of expression in the ventricular myocardium around birth. The present studies were performed to examine the expression pattern of the Cx40 protein in neonatal rat heart, and to examine the connexins present in cultures of ventricular myocytes obtained from those hearts. METHODS: Cryosections were made of hearts of 1-day-old Wistar rats. Cultures of ventricular myocytes obtained from these hearts by enzymatic dissociation were seeded at various densities (to obtain > 75, approximately 50%, and < 25% confluency) and cultured for 24, 48 or 96 h. Cx40 and Cx43 were detected by immunofluorescence and immunoblotting. RESULTS: Immunohistochemical stainings confirmed that gap junctions in the atrium and His-Purkinje system were composed of at least Cx43 and Cx40. From the subendocardium towards the subepicardium Cx40 expression gradually decreased, resulting in the sole expression of Cx43 in the subepicardial part of the ventricular wall. In ventricular myocytes cultured at high density (> 75% confluency) Cx43 and Cx40 immunoreactivity could be detected. In contrast to Cx43 immunolabeling which showed a homogeneous distribution pattern, Cx40 staining was heterogeneous, i.e. in some clusters of cells abundant labeling was present whereas in others no Cx40 staining could be detected. The pattern of Cx43 immunoreactivity was not altered by the culture density. In contrast, in isolated ventricular myocytes cultured at low density (< 25% confluency) the relative number of cell-cell interfaces that were Cx40 immunopositive decreased as compared to high density cultures (35 vs. 70%). Western blots did not reveal significant differences in the level of Cx40 and Cx43 expression at different culture densities. CONCLUSIONS: These results show that cultured ventricular myocytes retained typical features of the native neonatal rat ventricular myocardium with regard to their composition of gap junctions. This implicates that these cultures may serve as a good model for studying short-term and long-term regulation of cardiac gap junction channel expression and function. PMID- 10690314 TI - Use-dependent facilitation and depression of L-type Ca2+ current in guinea-pig ventricular myocytes: modulation by Ca2+ and isoprenaline. AB - OBJECTIVE: An increase in stimulation frequency can facilitate or depress cardiac Ca2+ current (ICa). The aim was to examine the Ca2+ dependence of these effects, to determine if facilitation is sustained, and to elucidate the mechanism by which isoprenaline modulates facilitation. METHODS: We examined the effects of increasing the stimulation frequency for 1 min, from 0.05 to 1 Hz, on ICa recorded from guinea-pig ventricular myocytes, using the whole-cell, voltage clamp technique. RESULTS: 1 Hz stimulation caused a facilitation of ICa that peaked in 5 s and was followed by depression towards the basal level. Metabolic inhibitors or replacement of extracellular Ca2+ with Ba2+ abolished facilitation without affecting depression, implying that they are independent processes and that facilitation required ATP and Ca2+. Subtraction of the depression observed in either condition, from the response to 1 Hz stimulation recorded under control conditions, revealed that ICa facilitation was well maintained during 1 Hz stimulation. Increased intracellular Ca2+ buffering reduced both phases of the response. Furthermore, varying the extracellular Ca2+ concentration ([Ca2+]o) revealed a Ca(2+)-dependent enhancement of depression and a bell-shaped dependence of facilitation on [Ca2+]o. Facilitation increased with [Ca2+]o up to 1 mM, then declined at higher concentrations due to partial masking by the overlaping depression. Isoprenaline produced concentration-dependent inhibition of facilitation and enhancement of depression when pipettes contained 2 mM EGTA, but not BAPTA. For an equivalent increase in ICa amplitude, the effects of isoprenaline and elevated [Ca2+]o on the response to 1 Hz stimulation were quantitatively the same. CONCLUSIONS: Facilitation is sustained during increased activity, but appears transient due to overlapping depression. Both responses are promoted by increased submembrane [Ca2+]. Isoprenaline appears to modulate facilitation and depression as a consequence of increased Ca2+ influx, rather than cAMP-dependent phosphorylation. The apparent block of facilitation by isoprenaline may result from masking by the enhanced depression. PMID- 10690315 TI - Stimulation of L-type Ca2+ current in human atrial myocytes by insulin. AB - OBJECTIVE: The L-type calcium current (ICa,L) in isolated human atrial myocytes was investigated as a possible target of insulin in the regulation of cardiac function. METHODS: Atrial myocytes were obtained from patients undergoing cardiac surgery. Using the whole-cell configuration of the patch-clamp technique, we investigated the stimulation of ICa,L by insulin in single human atrial myocytes. RESULTS: We found a dose-dependent stimulation of ICa,L by insulin at concentrations of 100 nM, 1 microM and 10 microM. Maximum stimulation of ICa,L over basal ICa,L was 140 +/- 12% (n = 11) at 10 microM insulin. The maximum conductance of ICa,L was increased by 10 microM insulin from 4.0 +/- 0.3 nS to 8.3 +/- 1.0 nS (n = 6). The stimulation of ICa,L by insulin was dose-dependent and reversible. Isoproterenol (10 nM) that stimulates ICa,L by 271 +/- 48% (n = 10) over basal ICa,L acted faster than insulin. The half-maximum stimulation of ICa,L by isoproterenol and insulin (10 microM) was reached after 31 +/- 2 s and 52 +/- 5 s, respectively. The insulin effect shown was totally reversed by acetylcholine (3 microM) which is known to inhibit adenylyl cyclase activity/cAMP production via Gi-proteins. Also, the selective insulin receptor tyrosine kinase inhibitor (hydroxy-2-naphthanelyl-methyl)phosphonic acid completely inhibited the insulin induced effect. CONCLUSION: Our data show that insulin stimulates the L type calcium current in isolated human atrial myocytes in a dose-dependent and reversible manner which appears to involve the insulin receptor tyrosine kinase. Insulin regulation of ICa,L in human atrial myocytes may be an interesting system for the analysis of the metabolic syndrome in man. PMID- 10690316 TI - Role of Ca2+ availability to myofilaments and their sensitivity to Ca2+ in myocyte contractile dysfunction in heart failure. AB - OBJECTIVE: Contractile function is depressed at the isolated myocyte level in heart failure (HF), which could result from the decreased availability of intracellular calcium ([Ca2+]i) to the myofibrils and/or the depressed sensitivity of myofilaments to [Ca2+]i. However, the cellular basis of contractile dysfunction remains unestablished. METHODS: We isolated left ventricular myocytes from dogs with rapid pacing-induced HF. Cell shortening and [Ca2+]i transients were measured by indo-1 fluorescence and the myofilament Ca2+ sensitivity was analyzed by the shortening-[Ca2+]i relation in intact myocytes as well as by the pCa tension relation in skinned cells. RESULTS: Peak cell shortening magnitude was depressed in HF, associated with a parallel decrease of [Ca2+]i transient amplitude. There was a significant positive correlation between these two variables (r = 0.71, P < 0.01). In contrast, myofibrillar sensitivity to Ca2+, determined by both intact and skinned myocytes, was comparable between control and HF. Further, there was no significant difference in Ca2+ sensitivity between control and HF even at shorter (1.8 microns) or longer (2.2 microns) sarcomere length. CONCLUSIONS: Using both intact and skinned cellular preparations, a potential defect in myocyte contractile function in HF was a reduction in Ca2+ availability to the myofilaments, rather than the inherent defects in myofilament sensitivity to Ca2+. PMID- 10690317 TI - Vasoconstriction by in situ formed angiotensin II: role of ACE and chymase. AB - OBJECTIVE: To assess the importance, for vasoconstriction, of in situ angiotensin (Ang) II generation, as opposed to ang II delivery to AT receptors via the organ bath fluid. METHODS: Ang I and II concentration-response curves in human and porcine coronary arteries (HCAs, PCAs) were constructed in relation to estimates of the clearances of Ang I and II (ClAngI, ClAngII) from the organ bath and the release of newly formed Ang II (RAngII) into the bath fluid. HCAs were from 25 heart valve donors (age 5-54 years), and PCAs from 14 pigs (age 3 months). RESULTS: Ang I- and II-evoked constrictions were inhibited by the AT1 receptor antagonist, irbesartan, and were not influenced by the AT2 receptor antagonist, PD123319. In HCAs Ang II was only three times more potent than Ang I, wheres, in the experiments with Ang I, comparison of ClAngI with ClAngII and RAngII indicated that most of the arterially produced Ang II did not reach the bath fluid. Also in PCAs Ang I and II showed similar potency. In HCAs both the ACE inhibitor, captopril, and the chymase inhibitor, chymostatin, inhibited Ang I evoked vasoconstriction, while only chymostatin had a significant effect on ClAngI. In PCAs Ang I-evoked vasoconstriction was almost completely ACE dependent. CONCLUSIONS: This study points towards the functional importance of in situ ACE- and chymase-dependent Ang II generation, as opposed to Ang II delivery via the circulation. It also indicates that functionally relevant changes in local Ang I-II conversion are not necessarily reflected by detectable changes in circulating Ang II. PMID- 10690318 TI - Maturation of the response to bradykinin in resistance and conduit pulmonary arteries. AB - OBJECTIVE: Immaturity of the endothelial-dependent relaxation is thought to be characteristic of the newborn pulmonary elastic arteries. In adulthood, the reactivity of different pulmonary arterial segments varies. Therefore, we investigated the presence of endothelial heterogeneity in perinatal porcine pulmonary arteries and compared it with the adult by studying the bradykinin-, substance P- and acetylcholine-induced relaxations in different arteries. METHODS: Three types of pulmonary arteries (large conduit elastic, distal branching and resistance-sized; in situ diameters 0.7-1.7, 0.3-0.5 and 0.1-0.2 mm, respectively) were isolated from lungs of adult (nine months), young (60-84 h), newborn (4 min) and near-term foetal pigs. They were mounted for isometric force recording, contracted first with K+ = 125 mmol/l (reference contraction). Cumulative concentration-response curves to acetylcholine, substance P or bradykinin were obtained from prostaglandin F2 alpha (30 mumol/l) precontracted vessels. The effects of captopril and O2(95 or 8%) were also determined. Experiments were terminated by adding 100 mumol/l papaverine, obtaining maximal relaxation, which was used for normalising relaxations. RESULTS: (i) Acetylcholine: In resistance arteries, relaxations were absent in the newborn and the adult. In conduit arteries, they were present from 60-84 h onward. (ii) Substance P: In resistance arteries, relaxations were only present in the adult. In the other two types of arteries, rudimentary relaxations were present from the mature foetal stage onward. (iii) Bradykinin: In resistance arteries, identical relaxations were present at all ages which, in the foetus and the adult, were insensitive to changes in O2 levels (95 to 8%). In conduit arteries, concentration-dependent relaxations were present from birth, increasing in amplitude with age and these were potentiated by captopril. Foetal conduit arteries relaxed to the single application of 0.1 mumol/l bradykinin, indicating age-dependent tachyphylaxis. CONCLUSIONS: (i) Bradykinin is unique among endothelium-dependent vasodilators in being able to relax all vascular segments, at all ages, subject to tachyphylaxis and bradykinin-breakdown but independent of the prevailing O2 concentration. (ii) Heterogeneity of the relaxations between conduit and resistance arteries is evident from the mature foetal stage onward. (iii) The type of agonist, the type of vessel and the age each independently determine the presence or absence of endothelial relaxations. Therefore, the perinatal pulmonary circulation is not immature with respect to endothelial dependent relaxation; rather, the nature of this process changes within the perinatal period and between birth and adulthood. PMID- 10690319 TI - A role for a glibenclamide-sensitive, relatively ATP-insensitive K+ current in regulating membrane potential and current in rat aorta. AB - OBJECTIVE: ATP-sensitive K+ channels have been classified based on their inhibition by cytoplasmic ATP. Recent evidence in vascular smooth muscle has suggested that these channels show weak sensitivity to intracellular ATP. However, it is not known whether these channels regulate the resting K+ conductance in vascular smooth muscles. Therefore, the aim of the present investigation was to characterize this current in rat aorta myocytes and to examine whether it contributes to setting the membrane potential. METHODS: The conventional and nystatin-permeablised whole cell patch clamp techniques were used to characterize the effect of glibenclamide on membrane potential and K+ current in enzymatically dispersed rat aorta myocytes. RESULTS: The mean resting potential measured in current clamp mode using the permeabilized patch approach was -54 +/- 5 mV (n = 8). Glibenclamide (10 microM) caused a reversible 24-mV depolarization in these cells. In symmetrical K+ (135 mM) solution an inward glibenclamide-sensitive (10 microM) current (-4.1 +/- 0.7 pA/pF; n = 5), hereafter termed Iglib, was observed at a membrane potential of -80 mV when cells held at -60 mV were ramped from -80 to +80 mV. In the absence of any nucleotide in the pipette solution, Iglib measured by the conventional whole-cell method was -23.69 +/- 4.65 pA/pF (n = 9). With 1 and 3 mM ATP in the pipette, the average current density was -25 +/- 6.3 pA/pF (n = 8), and -9.4 +/- 2.7 pA/pF (n = 9), respectively. In the absence of ATP, 1 mM GDP significantly (P < 0.01) increased Iglib (-44.8 +/- 8.4 pA/pF; n = 13). Inclusion of 1 mM ATP in the GDP-containing pipette solution had no significant effect on the current amplitude (-56.4 +/- 10.7 pA/pF; n = 7). Iglib fell to -11.0 +/- 2.9 pA/pF (n = 10) if 1 mM GDP and 3 mM ATP were present. In symmetrical K+, the Iglib observed in the presence of 1 mM ATP in the pipette was increased by more than two-fold in the presence of 10 microM levcromakalim. In PSS containing 5 mM K+, a significant glibenclamide sensitive current was observed at -45 mV membrane potential when cells dialyzed with 1 mM ATP were ramped between -80 to 30 mV. CONCLUSION: These results demonstrate that Iglib channels in rat aorta myocytes differ from classical KATP channels, being relatively insensitive to intracellular ATP. Iglib therefore appears to have an important role in contributing to the maintenance of the resting potential in rat aortic smooth muscle. PMID- 10690320 TI - Alterations in c-fos expression, cell proliferation and apoptosis in pressure distended human saphenous vein. AB - OBJECTIVES: Saphenous vein graft failures, resulting from thrombosis and the abnormal proliferation, migration and apoptosis of vascular smooth muscle cells (VSMC) are major limitations of coronary artery bypass surgery. We investigated whether surgical trauma of human saphenous vein induces the early response gene c fos and causes alterations in rates of proliferation and apoptosis. METHODS: Surgically prepared human vein consisted of distended (at 350 mmHg for 2 min) or non-distended segments of vein maintained in serum free RPMI at 37 degrees C and 5% CO2 for various time intervals. c-fos expression was detected by Northern analysis. Cell proliferation and apoptosis were determined by [3H]thymidine incorporation combined with proliferation cell nuclear antigen (PCNA) immunostaining and TUNEL, respectively. Labelling indices for proliferation and apoptosis were correlated with vessel was thicknesses. RESULTS: Control, freshly isolated vein expressed no c-fos. Surgically prepared vein synthesized c-fos 1 h following harvesting. There was a significant increase in c-fos in distended compared to non-distended vein. c-Fos protein increased in surgically prepared vein 24 h after harvesting. There was a significant increase in vascular cell proliferation in the non-distended compared to the distended vein: mean (S.E.M.) 1279 (218) vs. 863 (155) dpm/microgram DNA, P < 0.05, n = 6. In addition, the apoptotic index was significantly lower in the media of non-distended vs. distended vein 0.82 (0.2) vs. 5.5 (1.5), P < 0.05, n = 5. CONCLUSIONS: These findings demonstrate that surgical preparation of human saphenous vein increases expression of c-fos mRNA and apoptosis and reduces proliferation when compared with non-distended vein. These changes may influence the failure of saphenous vein grafts. PMID- 10690321 TI - Comment on "Interactions between endothelin-1 and the renin-angiotensin aldosterone system". PMID- 10690322 TI - NO in the cardiovascular system. PMID- 10690323 TI - Molecular control of nitric oxide synthases in the cardiovascular system. AB - Nitric oxide plays an important role in cardiovascular homeostasis. In this review, the regulation of the three nitric oxide synthase isoforms in the cardiovascular system are examined at molecular and cellular levels. In addition, recent information gleaned from the use of NOS knockout mice are discussed. PMID- 10690324 TI - Enzymatic function of nitric oxide synthases. AB - Nitric oxide (NO) is synthesised from L-arginine by the enzyme NO synthase (NOS). The complex reaction involves the transfer of electrons from NADPH, via the flavins FAD and FMN in the carboxy-terminal reductase domain, to the haem in the amino-terminal oxygenase domain, where the substrate L-arginine is oxidised to L citrulline and NO. The haem is essential for dimerisation as well as NO production. The pteridine tetrahydrobiopterin (BH4) is a key feature of NOS, affecting dimerisation and electron transfer, although its full role in catalysis remains to be determined. NOS can also catalyse superoxide anion production, depending on substrate and cofactor availability. There are three main isoforms of the enzyme, named neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS), which differ in their dependence on Ca2+, as well as in their expression and activities. These unique features give rise to the distinct subcellular localisations and mechanistic features which are responsible for the physiological and pathophysiological roles of each isoform. PMID- 10690325 TI - Signal transduction of eNOS activation. AB - Consistent with its classification as a Ca2+/calmodulin-dependent enzyme the constitutive endothelial nitric oxide (NO) synthase (eNOS) can be activated by receptor-dependent and -independent agonists as a consequence of an increase in the intracellular concentration of free Ca2+ ([Ca2+]i) and the association of the Ca2+/calmodulin complex with eNOS. Additional post-translational mechanisms regulate the activity of eNOS, including the interaction of eNOS with caveolin-1, heat shock protein 90 (Hsp90), or membrane phospholipids, as well as enzyme translocation and phosphorylation. In response to fluid shear stress the maintained production of NO by native and cultured endothelial cells is associated with only a transient increase in [Ca2+]i. In the absence of extracellular Ca2+ and in the presence of calmodulin antagonists, shear stress stimulates a maintained production of NO which is insensitive to the removal of extracellular Ca2+, but sensitive to tyrosine kinase inhibitors, Hsp90-binding proteins and phosphatidylinositol 3-kinase inhibitors. A pharmacologically identical activation of eNOS can be induced by protein tyrosine phosphatase inhibitors suggesting that the phosphorylation of eNOS, and possibly that of an associated regulatory protein(s), is crucial for its Ca(2+)-independent activation. PMID- 10690326 TI - Biological significance of endogenous methylarginines that inhibit nitric oxide synthases. AB - The guanidino-methylated arginine analogue NG monomethyl-L-arginine (L-NMMA) has been the standard nitric oxide synthase inhibitor used to evaluate the role of the L-arginine:nitric oxide pathway. However, L-NMMA and other methylated arginine residues are also synthesised in vivo by the action of a family of enzymes known as protein arginine methyltransferases. Proteolysis of proteins containing methylated arginine residues releases free methylarginine residues into the cytosol from where they may pass out of the cell into plasma. Of the three known methylarginine residues produced in mammals only asymmetrically methylated forms (L-NMMA and asymmetric dimethylarginine (ADMA)) but not symmetrically methylated arginine (symmetric dimethylarginine (SDMA)) inhibit nitric oxide synthase (NOS). We and others have proposed that endogenously produced asymmetrically methylated arginines may modulate NO production and that the accumulation of these residues in disease states may contribute to pathology. The activity of the enzyme dimethylarginine dimethylaminohydrolase that metabolises asymmetric methylarginines may be of critical importance in affecting NO pathways in health or disease. PMID- 10690327 TI - Interactions among ACE, kinins and NO. PMID- 10690328 TI - Interactions between NO and reactive oxygen species: pathophysiological importance in atherosclerosis, hypertension, diabetes and heart failure. AB - There is a growing body of evidence suggesting that numerous pathological conditions are associated with increased vascular production of reactive oxygen species. This form of vascular oxidant stress and particularly interactions between NO and oxygen-derived radicals represent a common pathological mechanism present in many so-called risk factors for atherosclerosis. Furthermore, reactive oxygen species seem to serve important cellular signalling mechanisms responsible for many of the features of vascular lesion formation. The mechanisms whereby vascular cells produce reactive oxygen species are only presently coming to light, and almost certainly will prove to be a focus for future therapies. PMID- 10690329 TI - Nitric oxide and coronary endothelial dysfunction in humans. PMID- 10690330 TI - Nitric oxide and the proliferation of vascular smooth muscle cells. PMID- 10690331 TI - NO and cardiac diastolic function. PMID- 10690332 TI - Regulation of cardiac beta-adrenergic response by nitric oxide. PMID- 10690333 TI - Nitric oxide, nitrates and ischaemic preconditioning. PMID- 10690334 TI - Inhaled nitric oxide in cardiology practice. AB - Inhaled nitric oxide allows selective pulmonary vasodilatation with rapidity of action. It is effective in the acute post-operative management of pulmonary hypertension in cardiac surgical patients and is also valuable in assessing the pulmonary vasodilator capacity in patients with chronic pulmonary hypertension. This review examines the current role of inhaled nitric oxide in cardiac medicine, discussing issues concerning its administration and toxicity, as well as a summary of clinical studies in cardiac patients. New roles, as a modifier of platelet and leukocyte function, post-thrombolysis and following lung transplantation are described. New agents and alternative therapies, which prolong pulmonary activity, are also discussed. PMID- 10690335 TI - Role of nitric oxide in the neural control of cardiovascular function. AB - The discovery in 1990 that nitric oxide (NO) acts as a neuromodulator within the central and peripheral nervous system triggered intensive research which considerably extended our understanding how this factor regulates cardiovascular functions. In addition to its direct effects on blood vessels NO has additional targets at all levels of the neural control of circulation. When not scavenged by hemoglobin, NO is relatively stable and diffuses over large distances (> 500 microns) so that one NO-producing cell can influence several thousands of adjacent cells in vivo. In different brain regions, NO and its metabolites have excitatory as well as inhibitory effects. The modulation of autonomic functions by these factors is therefore highly complex and often variable between the different levels from the brain to postganglionic nerve endings. This review is focused on the available evidence derived from animal studies and will summarize the current discussion about (i) the modulation of the generation of sympathetic and parasympathetic activities within the brain stem by NO; (ii) the actions of NO on cardiovascular reflexes and (iii) the role of NO as a modulator of autonomic functions within the target organs. Finally, the available evidence from human studies and some pathophysiological implications of altered NO mediated modulation of the neural control of circulation will be discussed. PMID- 10690336 TI - Inducible nitric oxide synthase and vascular injury. AB - The role nitric oxide (NO) plays in the cardiovascular system is complex and diverse. Even more controversial is the role that the inducible NO synthase enzyme (iNOS) serves in mediating different aspects of cardiovascular pathophysiology. Following arterial injury, NO has been shown to serve many vasoprotective roles, including inhibition of platelet aggregation and adherence to the site of injury, inhibition of leukocyte adherence, inhibition of vascular smooth muscle cell (VSMC) proliferation and migration, and stimulation of endothelial cell (EC) growth. These properties function together to preserve a normal vascular environment following injury. In this review, we discuss what is known about the involvement of iNOS in the vascular injury response. Additionally, we discuss the beneficial role of iNOS gene transfer to the vasculature in preventing the development of neointimal thickening. Lastly, the pathophysiology of transplant vasculopathy is discussed as well as the role of iNOS in this setting. PMID- 10690337 TI - Nitric oxide and penile erection: is erectile dysfunction another manifestation of vascular disease? AB - There is convincing evidence that the prevalence of erectile dysfunction is increased among men with ischaemic heart disease. This association may be attributed to the fact that both erectile dysfunction and ischaemic heart disease share similar risk factors (e.g. hypertension, dyslipidaemia, diabetes and smoking). Nitric oxide (NO) activity is adversely affected, in penile and vascular tissue, by these risk factors. It is therefore not surprising that a defect in NO activity is thought to play a role in the pathogenesis of both erectile dysfunction and ischaemic heart disease. We consider this evidence and propose that defective NO activity provides a unifying explanation for the association between these two conditions. Further research in this area may improve our understanding of the pathogenesis of cardiovascular diseases as a whole. PMID- 10690338 TI - 17 Beta-estradiol stimulates expression of endothelial and inducible NO synthase in rat myocardium in-vitro and in-vivo. AB - OBJECTIVES: NO production has been attributed to play a major role in cardiac diseases such as cardiac hypertrophy and cardiac remodeling after myocardial infarction which display significant gender-based differences. Therefore we assessed the effect of 17 beta-estradiol (E2) on estrogen receptor (ER) alpha and beta and endothelial and inducible NO synthase in neonatal and adult rat cardiomyocytes. METHODS: The presence of ER alpha and ER beta was demonstrated by immunofluorescence and western blot analysis as well as the expression pattern of inducible NO synthase (iNOS) and endothelial NOS (eNOS) in isolated cardiomyocytes from neonatal and adult rats. Furthermore, regulation of myocardial iNOS and eNOS expression by estrogen was evaluated in the myocardium from ovariectomized or sham-operated adult Wistar-Kyoto rats. RESULTS: Incubation with E2 led to translocalization of the ER into the nucleus and increased receptor protein expression. E2 stimulated expression of iNOS and eNOS in both neonatal and adult cardiac myocytes. Coincubation with the pure anti-estrogen ICI 182,780 inhibited upregulation of ER and NOS expression. In ovariectomized rats myocardial iNOS and eNOS protein levels were significantly lower compared to sham operated female animals. CONCLUSION: Taken together, these results show that E2 stimulates the expression of iNOS/eNOS in neonatal and adult cardiomyocytes in vivo and in-vitro. These novel findings provide a potential mechanism of how estrogen may modulate NOS expression and NO formation in the myocardium. PMID- 10690339 TI - Nitric oxide synthase expression and role during cardiomyogenesis. AB - OBJECTIVE: The aim of the present study was the investigation of the expression of NOS during cardiomyogenesis and its functional role. DESIGN: The qualitative and quantitative expression of NOS isoforms during different stages of cardiac development was evaluated using immunocytochemistry and dot blots, respectively. The functional relevance of NOS expression during cardiomyogenesis was investigated using the in vitro ES cell-differentiation model and selective pharmacological agents. RESULTS: On day 7.5 of embryonic development (E7.5) none of the NOS isoforms were expressed in the embryo, whereas the inducible (iNOS), as well as the endothelial (eNOS) isoforms were detected in the extraembryonic parts. In contrast, starting from E9.5 rat and murine embryos displayed prominent iNOS and eNOS expression. This was correlated with high expression of soluble guanylylcyclase (sGC) as well as high cyclic GMP (cGMP) content. During further development after E14.5 both, iNOS as well as eNOS, started to be downregulated and shortly prior to birth reduced staining for eNOS was found, whereas iNOS was hardly detectable. We further investigated whether NO plays a role for cardiomyogenesis, using in vitro ES cell-derived cardiomyocytes differentiating within embryoid bodies (EBs). The NOS expression pattern in these cells paralleled the one detected in vivo. We demonstrate that continuous incubation of EBs with the NOS inhibitors L-NMMA (2-10 mM) or L-NA (2-10 mM) for 4 to 9 days after plating resulted in a pronounced differentiation arrest of cardiomyocytes, whereas this effect could be reversed by coapplication of the NO-donor spermine NONOate (10 microM). CONCLUSIONS: Both, iNOS and eNOS isoforms are prominently expressed during early stages of cardiomyogenesis. Around E14.5 NOS expression starts to decline. Moreover, the NO-generation is required for cardiomyogenesis since NOS inhibitors prevent the maturation of terminally differentiated cardiomyocytes using the ES cell system. PMID- 10690340 TI - Repetitive myocardial stunning in pigs is associated with the increased expression of inducible and constitutive nitric oxide synthases. AB - OBJECTIVES: Nitric oxide (NO) has complex effects on myocardial function particularly following ischaemia-reperfusion. The goal of this study was to examine the result of repetitive myocardial stunning on myocardial NO release and expression of inducible (iNOS) and constitutive (eNOS) NO synthases. METHODS AND RESULTS: Propofol anaesthetised pigs underwent ten, 2-min episodes of circumflex artery occlusion (n = 6) or acted as sham operated controls (n = 4). Measurements of segment shortening demonstrated a fall in function in the ischaemic territory to 52.5 +/- 7.3% (mean +/- S.E.M.) of baseline shortening 30 min after the stunning stimulus, recovering to 92 +/- 8.7% 5.5 h later. Function remained stable in sham controls. The change in venous-arterial [NO] between baseline and 6 h reperfusion was found to be significantly different between the two groups (0.2 +/- 0.7 in stunned vs. -4.3 +/- 1.6 microM in shams; P < 0.02). Western blotting and band optical density used to compare tissue from stunned territory (S), non-stunned territory (IC) and sham control animals (SC) demonstrated this was associated with an increase in the expression of both iNOS (S: 93 +/- 13.4, IC: 37 +/- 2.4 and SC: 25 +/- 4 [arbitrary units], P < 0.01 and P = 0.031) and eNOS (S: 104 +/- 7.4, IC; 62.5 +/- 7.4 and SC; 75.7 +/- 0.6, P < 0.03 and P < 0.01) in stunned myocardium. Immunocytochemistry localised iNOS reactivity to vascular smooth muscle cells and cardiomyocytes in stunned tissue and eNOS reactivity to endothelial cells. CONCLUSION: Recovery from repetitive myocardial stunning is associated with the increased expression of both iNOS and eNOS and would be compatible with a protective role for both these enzymes. This finding has possible relevance for both the late window of ischaemic preconditioning and myocardial hibernation. PMID- 10690341 TI - Inducible nitric oxide synthase activation after ischemia/reperfusion contributes to myocardial dysfunction and extent of infarct size in rabbits: evidence for a late phase of nitric oxide-mediated reperfusion injury. AB - BACKGROUND: Ischemia/reperfusion (I/R) leads to the induction of inducible nitric oxide synthase. The present study investigated the effects of selective and continuous inhibition of iNOS on myocardial performance, infarct size and histomorphological changes after I/R in rabbits. METHODS AND RESULTS: Ischemia/reperfusion (I/R) was induced by occlusion of the circumflex coronary artery for 30 min followed by 48 h of reperfusion. Sham animals (group A) served as control. Three groups were subjected to I/R: (B) placebo; (C) aminoguanidine (AMG; 10 mg/kg bolus) given prior to and 48 h after I/R to test its acute effects; (D) AMG (300 mg/kg/day s.c.) to test effects of continuous treatment. Hemodynamics, myocardial blood flow, infarct size, iNOS activity, cGMP levels, immunohistochemical analysis of iNOS expression and AMG tissue levels were determined. Continuous AMG treatment improved myocardial performance (hemodynamics and blood flow) compared to placebo group. iNOS was highest in placebo-treated animals. AMG tissue levels were highest in tissues affected by I/R. Infarct size (% of the circumflex region) was significantly smaller in group D when compared to group B. CONCLUSIONS: This is the first study showing that activation of myocardial iNOS isozyme during 48 h of reperfusion contributes to a late phase of I/R-induced injury in rabbits. Selective and continuous modulation of iNOS by AMG over this time period exerts protective effects with respect to myocardial performance, coronary blood flow, cellular infiltration and reduction of infarct size; this may be a novel therapeutic approach in the clinical situation to limit irreversible myocardial injury associated with ischemia and reperfusion. PMID- 10690342 TI - Changes in extracellular pH mediate the chronotropic responses to L-arginine. AB - We have recently shown that exogenous nitric oxide (NO) elicits a positive chronotropic response by stimulating the hyperpolarization activated current, I(f). OBJECTIVE: To examine whether L-arginine (L-Arg) can mimic the chronotropic effect of NO by enhancing its endogenous production. METHODS: In spontaneously beating guinea pig atria we evaluated the heart rate (HR) response to increasing concentrations of L-Arg (1 mumol/l to 10 mmol/l), and compared it with that for D Arg or L-lysine (L-Lys) (all in free base (FB) or hydrochloride (HCl) formulation). RESULTS: L-ArgFB > 100 mumol/l caused a reversible dose-dependent increase in HR (peak effect +64 +/- 7 bpm at 10 mmol/l, P < 0.05, n = 8). However, a similar HR response occurred with D-ArgFB (n = 7) or L-LysFB (n = 6). All FB formulations increased the perfusate pH (peak [pH]o = 8.61 +/- 0.03). Although alkalinization can stimulate NO release from the endothelium, this is unlikely to have contributed to HR changes in our preparation, since neither NG methyl-L-arginine, (100-500 mumol/l, which per se reduced HR by 8 +/- 1%, P < 0.05, n = 9) nor NO scavenging (fresh 5% red blood cells, n = 9) caused a rightward shift of the concentration-response curve to L-ArgFB. Furthermore, as opposed to FB formulations, L-ArgHCl, D-ArgHCl or L-LysHCl > 1 mmol/l significantly decreased HR and [pH]o (n = 17). The chronotropic effects of L-ArFB or L-ArgHCl were reproduced by changing [pH]o with NaOH (n = 8) or HCl (n = 7), whereas the HR increase with L-ArgFB was prevented by clamping [pH]o at 7.42 +/- 0.07 (n = 10). CONCLUSIONS: In vitro, L-Arg can markedly affect HR through a pH mediated, NO-independent mechanism. Our data show that the opposing changes in [pH]o induced by different formulations of L-Arg can importantly confound the assessment of the biological effects of this amino acid. PMID- 10690343 TI - Metabolic vasodilation in the human forearm is preserved in hypercholesterolemia despite impairment of endothelium-dependent and independent vasodilation. AB - OBJECTIVE: Hypercholesterolemia has been shown to impair endothelium-mediated, nitric oxide (NO)-dependent responses to acetylcholine (ACh), serotonin, substance P and flow-mediated dilation. We have recently shown that NO contributes to metabolic vasodilation in the human forearm. We sought to determine whether metabolic vasodilation is impaired in healthy subjects with hypercholesterolemia. METHODS: We compared the forearm blood flow (FBF) responses to isotonic exercise, ACh and the endothelium-independent vasodilator sodium nitroprusside in young, otherwise healthy volunteers with hypercholesterolemia and controls before and after the NO inhibitor NG-monomethyl-L-arginine (L-NMMA). FBF was measured using venous occlusion plethysmography. Hypercholesterolemic (n = 20) and control (n = 20) subjects were age- and gender-matched. RESULTS: Total cholesterol (6.9 +/- 0.3 vs. 4.6 +/- 0.1 mmol/l, P < 0.0001), low density lipoprotein (4.9 +/- 0.4 vs. 2.7 +/- 0.1 mmol/l, P < 0.001) and triglyceride (1.3 +/- 0.2 vs. 0.8 +/- 0.1 mmol/l, P = 0.005) levels were higher in the hypercholesterolemic group. Basal FBF and resistance were similar in the two groups. Hypercholesterolemia impaired the peak FBF response to ACh (11.1 +/- 1.9 vs. 17.6 +/- 2.2 ml/100 ml/min, P = 0.03), and reduced the peak response to sodium nitroprusside (6.0 +/- 0.4 vs. 8.1 +/- 0.6 ml/100 ml/min, P < 0.01). However, hypercholesterolemia did not affect peak hyperemic FBF (13.1 +/- 1.0 vs. 13.2 +/- 1.0 ml/100 ml/min, P = 1.0) or the FBF volume repayment during the 1 or 5 min after exercise. Resting FBF was reduced by L-NMMA to a similar degree (by 33% vs. 40%, P = 0.17) in both groups. Although L-NMMA reduced peak hyperemic FBF (by 16% vs. 17%, P = 0.93) and the volume repaid after exercise in both groups, there were no differences between the two groups. CONCLUSIONS: Exercise-induced metabolic vasodilation is in part dependent on NO release. Hypercholesterolemia impairs NO-mediated vasodilation, but is not associated with a reduction in exercise-induced hyperemia. This may indicate that multiple compensatory mechanisms are operative in skeletal muscle metabolic vasodilation. PMID- 10690344 TI - L-arginine and L-NAME have no effects on the reendothelialization process after arterial balloon injury. AB - OBJECTIVE: Growth regulatory properties of nitric oxide (NO) in cultured endothelial cells is controversial. The aim of our study was to investigate the effect of L-arginine, the endogenous NO precursor, and L-NAME, an inhibitor of NO synthase on the reendothelialization process after angioplasty. METHODS: Fifty five New Zealand White rabbits underwent denudation of the left iliac artery. After injury the rabbits were randomized in three groups: L-arginine 2.25% (L arginine, n = 19); NG-nitro-L-arginine methyl ester 15 mg/kg/day (L-NAME, n = 19); and placebo (controls, n = 17). Treatment was solubilized in drinking water. Reendothelialization was evaluated at 4 weeks by macroscopic evaluation of Evans blue staining and endothelial-specific immunostaining (CD-31) on cross sections. Intimal hyperplasia was evaluated by morphometric analysis. RESULTS: Despite a significant increase in plasma arginine (P = 0.001) and a reduction in intimal hyperplasia (P = 0.003) with L-arginine, neither agent had a significant effect on reendothelialization at 4 weeks (controls = 36 +/- 4%, L-arginine = 43 +/- 3%, L-NAME = 33 +/- 4%; NS). CONCLUSION: These results suggest that, in spite of previously demonstrated effects on neointimal hyperplasia, the NO pathway does not influence the regrowth of macrovascular endothelial cells in vivo. PMID- 10690345 TI - Sympathectomy potentiates the vasoconstrictor response to nitric oxide synthase inhibition in humans. AB - OBJECTIVE: Nitric oxide exerts its cardiovascular actions at least in part by modulation of the sympathetic vasoconstrictor tone. There is increasing evidence that nitric oxide inhibits central neural sympathetic outflow, and preliminary evidence suggests that it may also modulate peripheral sympathetic vasoconstrictor tone. METHODS: To test this latter concept, in six subjects having undergone thoracic sympathectomy for hyperhydrosis, we compared the vascular responses to systemic L-NMMA infusion (1 mg/kg/min over 10 min) in the innervated and the denervated limb. We also studied vascular responses to the infusion of the non-nitric-oxide-dependent vasoconstrictor phenylephrine. RESULTS: L-NMMA infusion evoked a roughly 3-fold larger increase in vascular resistance in the denervated forearm than in the innervated calf. In the denervated forearm, vascular resistance increased by 58 +/- 10 percent (mean +/- SE), whereas in the innervated calf it increased only by 21 +/- 6 percent (P < 0.01, forearm vs. calf). This augmented vasoconstrictor response was specific for L-NMMA, and not related to augmented non-specific vasoconstrictor responsiveness secondary to sympathectomy, because phenylephrine infusion increased vascular resistance similarly in the denervated forearm and the innervated calf (by 24 +/- 7, and 29 +/- 8 percent, respectively). The augmented vasoconstrictor response was related specifically to denervation, because in control subjects, the vasoconstrictor responses to L-NMMA were comparable in the forearm and the calf. CONCLUSIONS: These findings indicate that in the absence of sympathetic innervation, the vasoconstrictor responses to nitric oxide synthase inhibition are augmented. PMID- 10690346 TI - Inducible nitric oxide synthase colocalizes with signs of lipid oxidation/peroxidation in human atherosclerotic plaques. AB - OBJECTIVE: Advanced human atherosclerotic plaques are characterized by the abundant presence of the autofluorescent non-soluble lipid pigment ceroid, consisting of oxidized lipoproteins. The aim of the present study was to examine the topographical and cellular distribution of inducible nitric oxide synthase (iNOS or NOS II) within different stages of atherosclerosis and its colocalization with ceroid deposits and nitrotyrosine. METHODS AND RESULTS: Different stages of atherosclerosis were studied by immunohistochemistry on whole mount longitudinal sections of carotid endarterectomy specimens. In the adaptive intimal thickening the predominant cell type were smooth muscle cells. The fatty streaks contained both smooth muscle cells and macrophages with an extremely low NOS II immunoreactivity. The advanced atherosclerotic plaques however, showed a very dense infiltration by macrophages, of which a subpopulation expressed NOS II as a vesicular immunoreactivity in their cytoplasm. These were mainly present around the necrotic core, in association with ceroid accumulation and nitrotyrosine. Fluorescence quenching microscopy showed the presence of NOS II on autofluorescent ceroid vesicles in the macrophages. Large extracellular ceroid granules were not NOS II immunoreactive. NOS II mRNA was detected by RT-PCR and the protein by Western blot in the plaque tissue but not in mammary arteries used as controls. CONCLUSION: Ceroid, nitrotyrosine and NOS II colocalized in late stages of atherosclerosis and were found around the necrotic core in the plaque. This could suggest that NOS II expression in macrophages is involved in oxidation and peroxidation of lipids, leading to ceroid formation. PMID- 10690347 TI - Effects of endothelin receptor antagonists and nitric oxide on myogenic tone and alpha-adrenergic-dependent contractions of rabbit resistance arteries. AB - Regulation of vascular contractions by endothelium-derived endothelin-1 (ET-1) and nitric oxide (NO) may vary depending on the stimulus. OBJECTIVES: To investigate if ET-1 receptor stimulation and NO contributed to a similar extent to the regulation of pressure- and alpha-adrenergic receptor (AR) agonist-induced smooth muscle contraction. METHODS: Rabbit mesenteric arteries (150-200 microns) were isolated, cannulated and pressurized. Changes in diameter were recorded as a function of the perfusion pressure (PP) or alpha-AR agonist addition at a PP of 60 mmHg. All experiments were performed in the presence of indomethacin (10 mumol 1(-1)). RESULTS: At a PP of 60 mmHg, myogenic tone (MT) developed to represent 17 +/- 1% of the minimal diameter. The magnitude of the MT was increased by 140% (P < 0.05) by the inhibition of NO production with N omega-nitro-L-arginine (L NOARG). Bosentan, an ETA/B receptor antagonist, and BQ 123, a selective ETA receptor antagonist, decreased (P < 0.05) MT either alone or in combination with L-NOARG by approximately 30%. Phenylephrine (Phe), an alpha 1-AR agonist, induced contraction; the sensitivity to Phe (pD2, 6.2 +/- 0.2) was unaffected by bosentan or BQ 123 alone but increased (P < 0.05) by L-NOARG (pD2, 7.3 +/- 0.5). Further addition of bosentan or BQ 123 restored the sensitivity to Phe to its control value. Oxymetazoline (OXY), an alpha 2-AR agonist, induced contraction; the sensitivity to OXY (pD = 2, 7.7 +/- 0.2) was unaffected by L-NOARG, bosentan or BQ 123. CONCLUSION: Our results indicate that pressure-induced tone is independently regulated by endothelium-derived NO and ET-1. In contrast, alpha 1 AR stimulation-induced tone is sensitive to ET-1 in the absence of NO, whereas occupation of alpha 2-AR mediates a contraction unregulated by the endothelium. PMID- 10690348 TI - NOS inhibition potentiates norepinephrine but not sympathetic nerve-mediated co transmission in resistance arteries. AB - OBJECTIVE: The in vitro interaction between sympathetic nerves and basal nitric oxide release was studied in a resistance artery, since these interact powerfully in large vessels. METHODS: The pharmacological interaction between L-NAME and vasoconstriction to field stimulation of sympathetic nerves or exogenous norepinephrine was studied in rabbit cutaneous resistance arteries in wire myographs. RESULTS: Relaxation of norepinephrine-induced tone by acetylcholine, but not sodium nitroprusside, was blocked by N omega-nitro-L-arginine methyl ester (L-NAME: 100 microM), indicating that the agonist-induced release of nitric oxide could oppose the vasoconstrictor effect of norepinephrine and confirming that L-NAME had no effect on endothelium-independent vasodilatation. L-NAME increased norepinephrine potency indicating basal NO release. With short bursts of electrical field stimulation purinergic transmission was dominant at low frequencies and adrenergic at high frequencies. L-NAME had no effect on nerve mediated responses, even after blocking the purinergic component with alpha,beta methylene ATP (3 microM), suggesting that the influence of spontaneously released nitric oxide does not extend to the vascular smooth muscle cells under adrenergic nervous control. CONCLUSION(S): This resistance artery exhibits a highly effective nitric oxide-mediated vasodilatation to acetylcholine. It has basal release of nitric oxide which antagonises exogenous norepinephrine. However, basal nitric oxide did not influence adrenergic nerve transmission, which contrasts with previous studies of larger arteries and veins. We speculate that in small resistance arteries there may be a spatial limitation to the zones of vascular smooth muscle influenced by the adrenergic nerves and by basal nitric oxide from the endothelium, respectively. The role of endogenous nitric oxide in modulating vascular tone may thus be less in resistance arteries than in conducting arteries or capacitance vessels and purinergic transmission appears to be particularly resistant. PMID- 10690349 TI - Nitroglycerin-mediated vasorelaxation is modulated by endothelial calcium activated potassium channels. AB - OBJECTIVE: Recent in vitro data suggest, large conductance calcium-activated K+ channels (BKCa) modulate the vascular response to nitric oxide (NO). The in vivo implications and the characteristics of this interaction are not clear. This study firstly investigates whether modulation of BKCa affects the vascular response to nitroglycerin (NTG)-derived NO in vivo and in the isolated heart and secondly examines the influence of endothelial BKCa on NTG-mediated vasodilation in vitro. METHODS: The hypotensive effect of NTG was measured in conscious, chronically catheterized rats during i.v. infusions of iberiotoxin (IbTX, a selective inhibitor of BKCa) or placebo. Similarly, NTG-induced flow-changes in the isolated perfused rat heart were examined before and after IbTX treatment (0.1 microM). Concentration-relaxation curves to NTG in the presence of various K+ channel modulating agents were performed in vitro on porcine coronary arteries with and without intact endothelium. RESULTS: I.v. infusion of IbTX reduced the in vivo hypotensive effect of NTG by 55% (before IbTX: 32.0 +/- 3.0 mmHg, vs. after IbTX: 14.5 +/- 3.2 mmHg, P < 0.05) and nearly abolished NTG-induced increase in coronary flow in the isolated perfused heart (P < 0.05). In vitro, this effect depended on an intact endothelium (endothelium intact segments; NTG: pD2 = 5.8 +/- 0.1, Emax = 97.6 +/- 3.2% vs. NTG + IbTX: pD2 = 4.9 +/- 0.2, Emax = 49.7 +/- 6.2%, P < 0.05; endothelium denuded segments; NTG: pD2 = 6.9 +/- 0.1, Emax = 104.0 +/- 1.4% vs. NTG + IbTX: pD2 = 6.7 +/- 0.1, Emax = 100 +/- 1.2%, P > 0.05). CONCLUSION: The results suggest, that modulation of endothelial BKCa significantly affects NTG-induced vasorelaxation in vitro, in the isolated perfused heart and in vivo. PMID- 10690350 TI - Role of basal nitric oxide synthesis in vasoconstrictor hyporeactivity in the perfused rat hindlimb after myocardial infarction: effect of captopril. AB - OBJECTIVES: The contribution of vascular changes to the development of heart failure is largely unknown. In the present study, we evaluated endothelial and vascular contractile function in the rat hindlimb vascular bed after myocardial infarction (MI), including the modulatory role of basal nitric oxide (NO) production and the effects of treatment with the angiotensin converting enzyme inhibitor captopril on vascular function. METHODS: MI was induced in male Wistar rats by ligation of the left coronary artery. Acetylcholine-induced dilatations were assessed in the ex vivo perfused hindlimb at various time points. At 2 and 5 weeks post-MI, vascular contractile function in the perfused hindlimb was assessed from resistance changes induced by 35 mM and 125 mM potassium (K+) and the maximum increase in resistance (delta Rmax, 125 mM K+ and 3 mg phenylephrine). Basal NO synthesis was blocked for 2 weeks with L-nitro-arginine methylester (L-NAME) in sham and MI rats and similar contractility experiments were performed. The effect of captopril treatment from 2 to 5 weeks post-MI on vasoconstrictor responses was also tested. RESULTS: Acetylcholine-induced dilatations in the presence of 10 microM indomethacin were not different between sham and MI rats. Vasoconstrictor responses to K+ and delta Rmax were reduced at 2 weeks after MI. This reduction in vasoconstrictor ability was similar to that seen in L-NAME-treated sham rats, while chronic L-NAME treatment did not affect vasoconstrictor reactivity in MI rats. Similarly, L-NAME induced an increase in mean arterial pressure in sham rats, but not in MI rats. At 5 weeks after MI, vasoconstriction to 125 mM K+ and delta Rmax were still reduced in MI rats; this response was however partially restored after captopril treatment. CONCLUSION: The development of vascular contractile hyporeactivity in the rat hindlimb after MI may be due to reduced basal NO production. Delayed treatment with captopril improves peripheral vascular contractile function in this setting. PMID- 10690351 TI - Estrogen receptor-alpha gene transfer into bovine aortic endothelial cells induces eNOS gene expression and inhibits cell migration. AB - OBJECTIVES: It has been suggested that estrogen may improve endothelial cell function to delay the onset of atherosclerosis in pre-menopausal females, though its mechanism of action is not fully understood. We examined the hypothesis that human estrogen receptor-alpha (ER alpha) gene transfection improves the endothelial cell function. METHODS: A replication deficient adenoviral vector was used to transfect the ER alpha gene into bovine aortic endothelial cells (BAEC) and a GFP gene containing vector was used as control. Expression of the eNOS gene was determined by Northern blot analysis and enzyme activity assay; cell migration was assayed using a Transwell apparatus; and tyrosine phosphorylation of FAK was estimated by Western blot analysis. RESULTS: ER alpha gene transfection of endothelial cells produced a 2-3-fold increase in eNOS mRNA and protein levels as well as a significant increase (P < 0.05) in NOS activity as measured by citrulline assay and nitrite accumulation in the media in response to bradykinin stimulation. Treatment of cells with estrogen blocking agent ICI 182780 inhibited eNOS induction in response to ER alpha transfection. ER alpha gene transfection significantly inhibited (P < 0.05) bFGF-induced chemotactic migration of endothelial cells but increased cell attachment to fibronectin, laminin, and type I and IV collagens. ER alpha gene transfer also inhibited bFGF stimulated tyrosine phosphorylation of FAK. CONCLUSION: Our results suggest that the atheroprotective effects of estrogen may in part be mediated by ER alpha induced upregulation of eNOS gene expression and maintenance of endothelial cell function and integrity. PMID- 10690352 TI - Gene transfer of endothelial nitric oxide synthase improves nitric oxide dependent endothelial function in a hypertensive rat model. AB - OBJECTIVE: We have shown previously that there is a relative nitric oxide deficiency at the level of vascular endothelium in the stroke-prone spontaneously hypertensive rat (SHRSP), a model of human essential hypertension, as compared to its normotensive reference strain Wistar Kyoto (WKY) rat. The aim of the current study was to investigate whether adenoviral-mediated gene transfer of an endothelial nitric oxide synthase (eNOS) cDNA (AdCMVeNOS) into carotid arteries of the SHRSP may improve endothelial function. METHODS: Enzyme activity of the recombinant eNOS protein encoded by AdCMVeNOS was tested using a Griess assay in endothelial cells in culture. Left carotid arteries of SHRSP were surgically isolated and exposed to either the AdCMVeNOS or control beta-galactosidase containing virus, (2 x 10(9) pfu/ml) ex vivo and in vivo. The vessels were harvested 24 h after surgery and analysed by Western blotting, immunohistochemistry and by examining endothelial function ex vivo. RESULTS: Cultured endothelial cells showed almost 100% transduction with both viruses and a dose response of eNOS expression showed a five-fold increase in nitrite production for AdCMVeNOS with no change for beta-galactosidase-containing virus. Western blotting demonstrated a significant increase of eNOS expression in vessels infused with AdCMVeNOS when compared to controls. Immunohistochemistry showed highly positive staining with monoclonal antibodies against eNOS in the intact endothelial cells of the AdCMVeNOS infused vessels. The areas under the curve of the concentration responses to phenylephrine (10(-9) to 3 x 10(-6) M) in the absence and presence of NG-nitroarginine methyl ester (100 microM) showed increased basal nitric oxide bioavailability in the carotid arteries infused with AdCMVeNOS compared to the control (n = 6 for each; P = 0.0069; 95% CI, 0.864 to 3.277). CONCLUSIONS: Our results show that AdCMVeNOS is an effective tool for vascular gene transfer and that it can improve endothelial NO availability in the SHRSP, a genetic model of essential hypertension and endothelial dysfunction. PMID- 10690354 TI - A pedigree-based study of mitochondrial D-loop DNA sequence variation among Arabian horses. AB - Through DNA sequence comparisons of a mitochondrial D-loop hypervariable region, we investigated matrilineal diversity for Arabian horses in the United States. Sixty-two horses were tested. From published pedigrees they traced in the maternal line to 34 mares acquired primarily in the mid to late 19th century from nomadic Bedouin tribes. Compared with the reference sequence (GenBank X79547), these samples showed 27 haplotypes with altogether 31 base substitution sites within 397 bp of sequence. Based on examination of pedigrees from a random sampling of 200 horses in current studbooks of the Arabian Horse Registry of America, we estimated that this study defined the expected mtDNA haplotypes for at least 89% of Arabian horses registered in the US. The reliability of the studbook recorded maternal lineages of Arabian pedigrees was demonstrated by haplotype concordance among multiple samplings in 14 lines. Single base differences observed within two maternal lines were interpreted as representing alternative fixations of past heteroplasmy. The study also demonstrated the utility of mtDNA sequence studies to resolve historical maternity questions without access to biological material from the horses whose relationship was in question, provided that representatives of the relevant female lines were available for comparison. The data call into question the traditional assumption that Arabian horses of the same strain necessarily share a common maternal ancestry. PMID- 10690353 TI - Highly efficient liposome-mediated gene transfer of inducible nitric oxide synthase in vivo and in vitro in vascular smooth muscle cells. AB - OBJECTIVE: The efficient introduction of regulatory genes into vascular smooth muscle cells (SMCs) is one of the most promising options for gene therapy of cardiovascular diseases. Cationic liposome-mediated gene transfer may become a favorable transfection technique with regard to patient's safety for in vivo administration. However, this method until now has its limitation in a low transfection efficiency. Therefore, the present study was designed to improve cationic liposome-mediated transfection of rabbit vascular SMCs in vitro and in vivo, in order to enhance transfection efficiency and present an optimized system which may offer a potential therapeutic benefit for in vivo application. METHODS AND RESULTS: Optimized lipofection of rabbit SMCs with the mammalian expression vector pE-N1 and the reporter gene green fluorescent protein resulted in a mean transfection efficiency of about 50%. The unique transfection of rabbit SMCs in vitro and in vivo with the inducible isoform of human nitric oxide synthase (NOSII), using the same vector, resulted in a successful transient transcription and translation of a functionally active human NOSII in rabbit SMC, persisting 5 6 days. We could further demonstrate that the transfection procedure and the transgene product did neither induce necrosis nor apoptosis under the conditions chosen and did not result in the induction of endogenous NOSII of transfected SMCs. CONCLUSION(S): These findings indicate potential therapeutic relevance for this nonviral gene transfer system for in vivo gene therapy for cardiovascular diseases. PMID- 10690355 TI - Construction and evaluation of a porcine bacterial artificial chromosome library. AB - A porcine bacterial artificial chromosome (BAC) library consisting of 103,488 clones has been constructed. The average insert size in the BAC vector was calculated to be 133 kb based on the examination of 189 randomly selected clones, indicating that the library contained 4.4 genome equivalents. The library can be screened by two-step PCR. The first screening step is performed on 22 superpools, each containing 4704 clones (49 x 96 well plates). In the second screening step, 49 plates comprising a superpool are arrayed in a 7 x 7 matrix and 4D-PCR is performed. Screening of the library superpools by PCR for 125 marker sequences selected from different regions of swine genome revealed 123 sequences, indicating that the library is not biased. Subsequent screenings (4D-PCR) were successfully applied for identification of clones containing each marker sequence. This porcine BAC library and the PCR screening system are useful for isolation of genomic DNA fragments containing desired sequences. PMID- 10690357 TI - Integration of the genetic and physical maps of the chicken macrochromosomes. AB - A large amount of genetic mapping information has been obtained in the chicken from the East Lansing, Compton and Wageningen reference populations. Physical mapping information has however, been more limited. We have mapped 14 new clones, both genetically and physically, and all 14 have been assigned to macrochromosomes. The orientation of linkage groups E01C01C11W01 (Chr 1), E06C02W02 (Chr 2), E02C03W03 (Chr 3), E05C04W04 (Chr 4), E07E34C05W05 (Chr 5), E11C10W06 (Chr 6), E45C07W07 (Chr 7) and E43C12W11 (Chr 8) has been established. Here we present integrated maps of the eight macrochromosomes and the Z chromosome of the chicken and correlate genetic with physical distances for chromosomes 1-3 and the Z sex chromosome. PMID- 10690356 TI - Chediak-Higashi syndrome mutation and genetic testing in Japanese black cattle (Wagyu). AB - Chediak-Higashi Syndrome (CHS) is an autosomal recessive disorder that affects several species including mice, humans, and cattle. Evidence based on clinical characteristics and somatic cell genetics suggests that mutations in a common gene cause CHS in the three species. The CHS locus on human chromosome 1 and mouse chromosome 13 encodes a lysosomal trafficking regulator formerly known as LYST, now known as CHS1, and is defective in CHS patients and beige mice, respectively. We have mapped the CHS locus to the proximal region of bovine chromosome 28 by linkage analysis using microsatellite markers previously mapped to this chromosome. Furthermore, we have identified a missense A:T-->G:C mutation that results in replacement of a histidine with an arginine residue at codon 2015 of the CHS1 gene. This mutation is the most likely cause of CHS in Wagyu cattle. In addition, we describe quick, inexpensive, PCR based tests that will permit elimination of the CHS mutation from Wagyu breeding herds. PMID- 10690358 TI - Identification and characterization of a new allele for the beta subunit of follicle-stimulating hormone in Chinese pig breeds. AB - During evaluation of follicle-stimulating hormone-beta (FSHB) expression in anterior pituitary glands by an RNase protection assay (RPA), the expected fragment of 205 nucleotides at positions 759-963 was not detected in one boar that had moderate plasma and pituitary FSH concentrations. After subcloning and sequencing, mRNA from this boar lacked an 11-bp fragment (5'-CATTTGGAAAC-3') at nucleotide positions 807-817 of the 3'-untranslated region (3'-UTR, D allele). Wild-type FSHB (WT allele) was present in pituitary RNA and genomic DNA in both Meishan (MS) and White Composite (WC) pigs; whereas the D allele was present only in MS pigs (P < 0.01; 5/6 MS vs. 0/6 WC). Also, we found the D allele in five other Chinese breeds but absent in ten American Landrace, 11 Yorkshire and 17 Berkshire pigs. Additionally, the D allele had one silent nucleotide change in the coding region plus six, single nucleotide changes in the 3'-UTR. PMID- 10690359 TI - Microsatellite analysis reveals substantial genetic differentiation between contemporary New World and Old World Holstein Friesian populations. AB - Genotypic data from 39 microsatellite loci typed in 211 animals were used to assess the genetic differentiation between Old World and New World Holstein Friesian cattle populations. Gene diversities were similar in all five Holstein Friesian populations surveyed, ranging from 0.43 to 0.48. A tree of individuals based on the proportion of shared alleles indicated a clear distinction between Old World and New World Holstein Friesian populations. Similarly, genetic differentiation between populations, as measured by FST, was highly significant. Using the split decomposition method, we were able to visualize the significant introgression of New World Holstein Friesian into European Holstein Friesian populations. PMID- 10690360 TI - The genetic structure of Spanish Celtic horse breeds inferred from microsatellite data. AB - Partition of the genetic variability, genetic structure and relationships among seven Spanish Celtic horse breeds were studied using PCR amplification of 13 microsatellites on 481 random individuals. In addition, 60 thoroughbred horses were included. The average observed heterozygosity and the mean number of alleles were higher for the Atlantic horse breeds than for the Balearic Islands breeds. Only eight percentage of the total genetic variability could be attributed to differences among breeds (mean FST approximately 0.08; P < 0.01). Atlantic breeds clearly form a separate cluster from the Balearic Islands breeds and among the former only two form a clear clustering, while the rest of Atlantic breeds (Jaca Navarra, Caballo Gallego and Pottoka) are not consistently differentiated. Multivariate analysis showed that Asturcon populations, Losina and Balearic Islands breeds are clearly separated from each other and from the rest of the breeds. In addition to this, the use of the microsatellites proved to be useful for breed assignment. PMID- 10690361 TI - Biochemical, molecular and physiological characterization of a new beta-casein variant detected in Korean cattle. AB - There are seven known genetic variants of bovine beta-casein (beta-CN)--A1, A2, A3, B, C, D and E. In this study, we identified a new genetic variant (named beta CN H) which migrates slower than the other variants in acidic starch gel electrophoresis. We confirmed through protein and DNA sequence analyses that the H variant differs at five residues from the A2 sequence: Arg25/Cys, Leu88/Ile, Gln117/Glu, Glu175/Gln and Gln195/Glu. Of these substitutions the 25th residue was contained in the casein phosphopeptide (CPP) region. In rats, calcium solubilizing effect of the CPP of bovine variant H was increased by approximately 23% compared with that of the CPP of non-H. Using extensive Korean Bos taurus pedigrees, we confirmed that beta-CN H was controlled by a codominant allele. PMID- 10690362 TI - Nomenclature for factors of the dog major histocompatibility system (DLA), 1998: first report of the ISAG DLA Nomenclature Committee. AB - A Nomenclature committee for Factors of the Dog Major Histocompatibility System or Dog Leukocyte Antigen (DLA) has been convened under the auspices of the International Society for Animal Genetics (ISAG) to define a sequence based nomenclature for the genes of the DLA system. The remit of this committee includes: assignment of gene names rules for naming alleles assignment of names to published alleles assignment of names to new alleles rules for acceptance of new alleles DLA Nomenclature Committee, rules for acceptance, DLA genes and alleles, sequence based nomenclature. PMID- 10690363 TI - Development and characterization of expressed sequence tags for the turkey (Meleagris gallopavo) genome and comparative sequence analysis with other birds. AB - Twenty-one randomly selected clones from a turkey (Meleagris gallopavo) pituitary complementary DNA (cDNA) library were sequenced to develop expressed sequence tags (ESTs) for this economically important avian species whose genome is among the least understood. Primers specific for the ESTs were used to produce amplicons from the genomic DNA of turkey, chicken (Gallus gallus), guinea fowl (Numidia meleagris), pigeon (Columba domestica), and quail (Corturnix japonica). The amplicons were sequenced and analyzed for sequence variation within- and similarity among-species and with GenBank database sequences. The proportion of shared bases between the turkey sequence and the consensus sequence from each of the other species ranged from 72% to 93% between turkey and pigeon and quail and between turkey and chicken, respectively. The total number of single nucleotide polymorphisms (SNPs) observed ranged from 3 in quail to 18 in chicken out of 4898 and 5265 bases analyzed, respectively. The most frequent nucleotide variation observed was a C-->T transition. Linkage analysis of one such SNP in the backcross progeny of the East Lansing reference DNA panel, localized TUS0005, the chicken sequence derived from primers specific for turkey TUT2E EST, to chromosome 4. The ESTs reported, as well as the SNPs may provide a useful resource for ongoing efforts to develop high utility genome maps for the turkey and chicken. The primers described can also be used as a tool in future investigations directed at further understanding the biology of the guinea fowl, pigeon and quail and their relatedness to the turkey. PMID- 10690364 TI - Tth111I PCR/RFLP marker in the canine rod transducin alpha (GNAT1) gene. PMID- 10690365 TI - Ten equine microsatellite loci: TKY25, TKY26, TKY27, TKY28, TKY29, TKY267, TKY268, TKY269, TKY270 and TKY271. PMID- 10690366 TI - Physical assignment of the porcine erythropoietin receptor gene to SSC2. PMID- 10690367 TI - Identification of a MaeI RFLP in the insulin-like growth factor-1 (IGF1) gene of swamp buffaloes (Bubalus b. bubalis kerebau). PMID- 10690368 TI - A DNA polymorphism in the bovine c-kit gene. PMID- 10690369 TI - Variable microsatellites in the Pacific oyster Crassostrea gigas and other cupped oyster species. PMID- 10690370 TI - Four microsatellite markers in the Japanese flounder, Paralichthys olivaceus. PMID- 10690371 TI - Two bovine dinucleotide repeat polymorphisms: RM084 and RM171. PMID- 10690372 TI - Four highly polymorphic dinucleotide microsatellites in rainbow trout (Oncorhynchus mykiss). PMID- 10690373 TI - A diallelic short tandem repeat (CCCCG)4 or 5, located in intron 1 of rabbit alpha-globin gene. PMID- 10690374 TI - South American camelid microsatellite amplification in Camelus dromedarius. PMID- 10690375 TI - A single nucleotide (T-->G) polymorphism within intron 23 of the canine BRCA1 gene. PMID- 10690376 TI - Seven novel cosmid-derived canine microsatellites. PMID- 10690377 TI - Characterization of ten equine dinucleotide microsatellite loci: NVHEQ21, NVHEQ54, NVHEQ67, NVHEQ70, NVHEQ75, NVHEQ77, NVHEQ79, NVHEQ81, NVHEQ82 and NVHEQ83. PMID- 10690378 TI - New mutation in exon 2 of the bovine leptin gene. PMID- 10690379 TI - 'Old Kerr's almanac'. PMID- 10690380 TI - The control of acute menopausal symptoms in breast cancer survivors. PMID- 10690381 TI - Recurrent squamous-cell carcinoma of the head and neck: overview of current therapy and future prospects. AB - Locoregional recurrence is the most common cause of failure after head and neck cancer surgery. It is a disease which causes significant morbidity especially on speech and swallowing. There are many different treatments available including surgery, reirradiation and chemotherapy. However, none of these have produced any significant survival benefit. Because of this, there has been considerable interest in the development of new biological therapies such as gene therapy and immunotherapy for this disease. The objectives of this article are to provide an overview of the currently available therapies for recurrent head and neck cancer including gene therapy and immunotherapy. Prevention of recurrent disease by the detection and treatment of minimal residual disease is also discussed. PMID- 10690382 TI - A pilot trial assessing the efficacy of paroxetine hydrochloride (Paxil) in controlling hot flashes in breast cancer survivors. AB - BACKGROUND: Many breast cancer survivors suffer debilitating hot flashes. Estrogen, the drug of choice in perimenopausal women, is generally not recommenced to breast cancer survivors. Nonhormonal treatments are mostly disappointing. Anecdotal reports in our institution suggested that the selective serotonin-reuptake inhibitor, paroxetine hydrochloride, might be efficacious in alleviating hot flashes. PATIENTS AND METHODS: Thirty women with prior breast cancer who were suffering at least two hot flashes a day entered a single institution pilot trial to evaluate paroxetine's efficacy in reducing the frequency and severity of hot flashes. After completing daily diaries for one week on no therapy, the women received open-label paroxetine, 10 mg daily for one week, followed by four weeks of paroxetine, 20 mg daily. The women completed hot flash daily diaries throughout the study period, and a health-related symptom assessment questionnaire and a quality-of-life rating scale in the first and sixth week of the study. RESULTS: Twenty-seven women completed the six-week study period. The mean reduction of hot flash frequency was 67% (95% confidence interval (95% CI): 56%-79%). The mean reduction in hot flash severity score was 75% (95% CI: 66%-85%). There was a statistically significant improvement in depression, sleep, anxiety, and quality of life scores. Furthermore, 25 (83%) of the study participants chose to continue paroxetine therapy at the end of study. The most common adverse effect was somnolence, resulting in drug discontinuation in two women, and dose reduction in two women. One woman discontinued drug due to anxiety. CONCLUSIONS: Paroxetine hydrochloride is a promising new treatment for hot flashes in breast cancer survivors, and warrants further evaluation in a double-blind randomized placebo-controlled trial. PMID- 10690383 TI - A clinical model for quality of life assessment in cancer patients receiving chemotherapy. AB - BACKGROUND: The pattern of symptoms experienced by cancer patients during chemotherapy is very complex. Consequently, quality of life (QOL) assessment has to be carefully planned to capture clinically relevant changes. PATIENTS AND METHODS: A clinical model of changes in symptoms experienced by symptomatic metastatic patients during several courses of chemotherapy has been developed. The model differentiates cancer-related symptoms, acute side-effects, chronic side-effects and symptoms not related to cancer. The model was used to predict changes in each of these four symptom groups. Three time points were selected (post-cycle 2, pre-cycle 3, post-cycle 5) and an appropriate window around each time point was set. The model predictions were tested empirically with 56 patients with advanced ovarian cancer who completed the EORTC QLQ-C30 plus disease specific items during a six-cycle course of chemotherapy. RESULTS: The changes observed in the sample were in accordance with the changes predicted by the clinical model. Results from patients who did not complete the questionnaire within the specified time windows tended to dilute the findings from the group who did. CONCLUSIONS: A clinical model is useful in the planning of QOL assessments in order to capture clinically relevant effects. Such models also facilitate the interpretation of QOL studies, particularly when cyclic short-term effects and chronic side-effects are overlaid on disease symptoms, as is the case with chemotherapy for cancer. PMID- 10690384 TI - Assessment of hospitalised cancer patients' needs by the Needs Evaluation Questionnaire. AB - BACKGROUND: Cancer disease modifies the order and the nature of needs connected with the state of health. The aim of this study was to evaluate the informative, psychological, social and practical needs of hospitalised cancer patients by means of the Needs Evaluation Questionnaire (NEQ), an instrument designed concisely for the convenience of patients and medical staff. PATIENTS AND METHODS: Different samples of consecutive hospitalised cancer patients were involved in the various phases of designing the instrument: 30 patients for items identification, 101 patients for completeness and acceptability evaluation, 423 patients for construct validity and prevalence of needs; content and reliability analysis were performed on 2 subsamples of, respectively, 60 and 88 of the patients from the last sample. RESULTS: The validation analysis showed rather good reliability, structure validity and internal consistency of the questionnaire. The prevalence analysis showed that the most common needs were: 'more information about my future conditions' (74%); 'more information about my diagnosis' (56%); 'more information about the exams I am undergoing' (52%); 'more explanations on treatments' (51%); 'to have a better dialogue with clinicians' (57%); 'better services from the hospital' (bathrooms, meals, cleaning) (56%). CONCLUSIONS: The NEQ, self-completed by patients, has proven to be a useful clinical tool for obtaining a systematic and undistorted overview of the principal needs with respect to the state of health of patients. This instrument, which can also be administered by persons not belonging to the health care system such as volunteers, and inserted into the patients' hospital charts, could be used by the medical staff to identify the real needs of patients at an early stage. PMID- 10690385 TI - Patient participation in medical decision-making: a French study in adjuvant radio-chemotherapy for early breast cancer. AB - BACKGROUND: Shared decision-making is increasingly advocated as an ideal model. However, very few studies have tested the feasibility of giving patients the opportunity to participate in the choice of treatment. PATIENTS AND METHODS: Women, with non-metastatic breast cancer, eligible for non-intensified adjuvant chemotherapy attending our hospital were proposed two administrations of chemotherapy and radiotherapy: a sequential and a concomitant one. Two patient questionnaires were used to elicit motivations for their choice and their degree of comfort with the process of decision-making and one questionnaire to test physicians' ability to predict patients' choice. RESULTS: Participation rate in the study was 75.3% (n = 64). Majority (64%) of patients chose the concomitant treatment. Multivariate analysis revealed that patients with a lower level of education, who discussed the choice with social circle, and who most feared side effects were more likely to choose the sequential treatment. Physicians were able to predict patients' choice in 66% of cases. 89% of patients declared that they were fully satisfied with having participated in the choice of treatment and 79% supported shared decision-making. CONCLUSIONS: Results are in favour of promoting active participation of cancer-patients in medical decision-making. The adequate degree of such participation remains however to be elicited and tested for therapeutic choices implying more difficult trade-offs between quantity and quality of life. PMID- 10690386 TI - Mature B-cell lymphoma/leukemia in children and adolescents: intergroup pathologist consensus with the revised European-American Lymphoma Classification. AB - BACKGROUND: The Revised European-American Lymphoma (R.E.A.L.) Classification criteria were evaluated in the international protocol FAB LMB 96 Treatment of Mature B-cell Lymphoma/Leukemia: A SFOP LMB 96/CCG-5961/UKCCSG NHL 9600 Cooperative Study. This includes B-lineage lymphomas: Burkitt's lymphoma (including ALL-L3); high-grade B-cell lymphoma, Burkitt-like; diffuse large B cell lymphoma (excluding anaplastic large cell Ki-1 lymphoma). PATIENTS AND METHODS: Cases were independently reviewed by eight hematopathologists from the three cooperative national groups (two SFOP, two CCG, four UKCCSG), without prior discussion of classification criteria or guidelines for case rejection. Consensus diagnosis was determined by each national cooperative group, and final consensus diagnosis established when at least two national consensus diagnoses were in agreement, or following group agreement at a multiheaded microscope. RESULTS: Two hundred eight cases were reviewed, with final consensus diagnosis established in two hundred three. The percent agreement of each group's national consensus diagnosis with final consensus diagnosis was 86%, 86% and 71%. The percent agreement of the group's national consensus diagnosis with final consensus diagnosis for Burkitt's and diffuse large B-cell lymphoma were 88% and 80%, respectively, but only 42% for Burkitt-like lymphoma. CONCLUSIONS: International panel review of mature B-cell lymphoma/leukemia in children and adolescents highlighted difficulties in subclassification, particularly with Burkitt-like, which is a 'provisional entity' in the R.E.A.L. Classification. The absence of previous discussion of classification and guidelines for case rejection may in part explain the discrepancy between pathologists. These results underline that morphology may need to be complemented by other studies, such as molecular genetic and cytogenetics, to discriminate between the mature B-cell lymphomas. PMID- 10690388 TI - Hodgkin's disease: correlation between causes of death at autopsy and clinical diagnosis. AB - PURPOSE: The causes of mortality in Hodgkin's disease patients are insufficiently known. Autopsy study is the fundamental procedure in the investigation of these causes. The present study analyzes the autopsies performed in a series of patients diagnosed as having Hodgkin's disease, determining the cause of death in each case and comparing the premortem clinical data and the postmortem findings. PATIENTS AND METHODS: A total of 486 patients diagnosed as having Hodgkin's disease between 1967 and 1996 were assessed. Autopsy was performed in 40 of the 144 deceased patients (28%). We reviewed the pathological findings, effects of treatment, discordance between the clinical diagnosis and the outcome of autopsy, and cause of death in each case. RESULTS: The most common clinical causes of death in those patients with autopsy study were tumor progression (37%) and infections (43%) in those patients with autopsy study. The rate of discordance between the clinical and autopsy diagnoses in this study was 43%. The most frequent location of residual Hodgkin's disease was in the lymph nodes. CONCLUSIONS: Autopsy study in Hodgkin's disease confirms a high rate of discrepancy between final clinical diagnosis and postmortem lesions despite advances in diagnostic methods. Autopsy revealed causes of death directly related to the treatment, as well as some lesions directly related to patient death and secondary to treatment. Infectious processes are likely to remain undetected and their symptoms can mimic tumor progression. PMID- 10690387 TI - Relapses of childhood anaplastic large-cell lymphoma: treatment results in a series of 41 children--a report from the French Society of Pediatric Oncology. AB - PURPOSE: To study response to chemotherapy and the outcome of children treated for a relapsed anaplastic large-cell lymphoma (ALCL) and to evaluate the role of bone marrow transplantation (BMT) in these patients. PATIENTS AND METHODS: Clinical data concerning the 41 relapses that occurred in 119 patients with ALCL enrolled in 3 consecutive studies since 1975 were analysed. First-line treatment consisted of intensive chemotherapy according to the COPAD protocol for the first series of 12 patients treated between 1975 and 1989 and to the SFOP (French Society of Pediatric Oncology) HM protocols for the 30 patients treated between 1989 and 1997. Twenty-eight patients were treated with CV(B)A (CCNU, vinblastine, ara-C with or without bleomycin), and the others with miscellaneous protocols for recurrent disease. Fifteen patients underwent autologous BMT and 1 allogeneic BMT while in CR2. RESULTS: Thirty-six of forty-one (88%) patients achieved CR2. With a median follow-up of 5 years, 12 patients died, 9 of their disease and 29 patients are alive in CR2 (20 patients), CR3 (5 patients), CR4 (2 patients), CR5 (1 patient) or CR6 (1 patient). Overall and disease-free survival are respectively 69% (53%-82%) and 44% (29%-61%) at three years. In univariate analysis, patients treated with ABMT while in CR2 did not appear to have a better outcome than the other. Remarkably, a long-lasting remission was obtained in 8 of 13 patients treated with weekly vinblastine for a relapse including 6 relapses occurring after ABMT. CONCLUSIONS: Relapsed ALCL are highly chemosensitive but over 40% of the patients experience several relapses. Prolonged conventional chemotherapy based on vinblastine might, in some cases, be as efficient as short intensive treatment with ABMT. PMID- 10690389 TI - Second primary malignancies following the treatment of early stage ovarian cancer: update of a study by the National Cancer Institute of Canada--Clinical Trials Group (NCIC-CTG). AB - BACKGROUND: Ovarian cancer is the leading cause of death from gynecological malignancies and the fourth most frequent fatal malignancy in women. Despite improved surgical techniques as many as 20% of women with early stage disease will eventually relapse and die from their disease. The post-operative management of these women remains controversial. Here we present the long term follow-up data of our previously published study, as well as the incidence of second primary malignancies in these women. PATIENTS AND METHODS: Two hundred fifty seven eligible patients with stage I, IIA 'high risk' ovarian carcinoma and IIB, IIIO (disease confined to pelvis) were randomized to either whole abdominal radiotherapy 2.250 rads in ten fractions (107 patients), melphalan 8 mg/m2/d x 4 weeks x 18 courses (106 patients) or intraperitoneal chromic phosphate 10-20 mCi (44 patients). All patients were initially treated with pelvic radiotherapy. RESULTS: Overall survival estimates at 10 years were: 45% in the whole abdominal radiotherapy arm; 49% in the melphalan arm and 50% in the intraperitoneal chromic phosphate arm (P = 0.30). Relapse-free survival estimates at 10 years were: 50% in the whole abdominal radiotherapy arm, 62% in the melphalan arm and 51% in the chromic phosphate arm (P = 0.147). Long term follow-up has not demonstrated a significant difference between treatment arms. Second primary malignancies developed in 29 women (11%) after 2,229 person years of follow-up. This compares to 18.7 second primary malignancies which would have been expected in this group of age-matched controls and was statistically significant (P = 0.018). There was no significant difference in the total number of second primary malignancies between treatment arms. Melphalan appeared to be associated with an increased risk of developing leukemia/myelodysplastic syndrome compared to the whole abdominal radiotherapy arm (P = 0.06). CONCLUSIONS: Long-term follow-up has not demonstrated a significant difference in overall or disease free survival between treatment arms. An excess of second primary malignancies (35%) was observed suggesting that lifelong surveillance is required in this population. Further research with newer treatment programs are needed to improve the cure rates in this population. PMID- 10690392 TI - BRCA2 germ-line mutations in Spanish male breast cancer patients. AB - BACKGROUND: Mutations in the BRCA2 gene account for the majority of the families with male and female breast cancer cases, and a number of BRCA2 mutations have been reported in males with breast cancer. The aim of this study was to characterise BRCA2 germ-line mutations in Spanish male breast cancer patients. PATIENTS AND METHODS: We screened DNA from 11 affected men and 6 women with breast cancer (BC) who had an affected male relative (father or brother). Exons 2 9 and 12-27 were screened by SSCP, and exons 10 and 11 were screened by PTT. PCR products with a variant band were sequenced. RESULTS: Three BRCA2 frameshift mutations were identified (17.6%): the 3374delA in codon 1049 (exon 11), 6857delAA in codon 2010 (exon 11), and 9254delATCAT in codon 3009 (exon 23). These mutations were present in patients with affected first-degree relatives (3 of 9, 33%). The proportion of male patients with a family history of BC in at least one first-degree relative was 53%. CONCLUSIONS: There is an association between BRCA2 mutations and male breast cancer, especially in those with a family history of BC. The high prevalence of BRCA2 mutations among males should be considered when estimating risk for female relatives. All new male cases of BC should be regarded as being possibly inherited and should be fully investigated. PMID- 10690391 TI - A dose-finding study of gemcitabine and vinorelbine in advanced previously treated malignancies. AB - PURPOSE: Gemcitabine and vinorelbine are active drugs with broad spectrum of activity and manageable toxicity in clinical trials. The aims of this study were to describe the toxicity, to determine the dose-limiting toxicity, and to define the doses of gemcitabine and vinorelbine to be recommended for phase II studies in patients with advanced cancers. PATIENTS AND METHODS: Drugs were given as 30 min infusions on day 1 and 8 (vinorelbine before gemcitabine) every 3 weeks. Thirty-six patients (male:female ratio 25:11; mean age 54, PS > 60) were treated including 1 retroperitoneal sarcoma, 7 head and neck, 10 lung, 4 thyroid, 6 pancreatic, 1 bladder, 2 ovary, 2 gastric, 1 rectum, 1 unknown primary, and 1 renal cell carcinoma. Doses of gemcitabine/vinorelbine ranged from 800/20 mg/m2 to 1500/30 mg/m2. RESULTS: The dose-limiting toxicity was neutropenia. A transient grade 2-3 elevation of transaminases was frequently observed at several dose-levels, although this toxicity did not appear to be dose dependant and was reversible at day 21 before the next cycle. Other toxicities were mild and easily manageable, consisting of fatigue and flu-like syndromes. Since the MTD was not reach at the higher dose-level, the recommended dose level of the gemcitabine vinorelbine combination was 1500/30 mg/m2. One toxic death due to hematologic toxicity was reported in a heavily pretreated patient who underwent prior chemotherapy and pelvic radiotherapy. A total of 12 patients were treated at the recommended dose level which was associated with grade 3-4 neutropenia in 3 of 12 patients and in 22.9% of cycles. CONCLUSIONS: This study estimates that the recommended dose for phase II studies of gemcitabine-vinorelbine is 1500/30 mg/m2 at day 1 and 8 every three weeks. A careful monitoring of the hematologic toxicity is recommended in heavily pretreated patients and in patients who received pelvic radiotherapy. Partial responses observed in a patient with an advanced cisplatin-5-fluorouracil-resistant pancreatic adenocarcinoma and in a patient with mesothelioma support further evaluation of this combination in patients with tumors refractory to classical antitumor agents. PMID- 10690390 TI - Liposomal vincristine in relapsed non-Hodgkin's lymphomas: early results of an ongoing phase II trial. AB - OBJECTIVE: Vincristine is an active agent in lymphomas, but is often neurotoxic, and the resulting dose reductions have been associated with lower remission and survival rates in Hodgkin's disease. Liposomal vincristine (Onco-TCS) has prolonged half-life, reaches higher concentration in tumors and lymph nodes than in nerves, and administered at full doses appears to be less neurotoxic, and more active then free vincristine in mice bearing L-1210 and P-388 leukemias. We therefore explored its activity in relapsed non-Hodgkin's lymphomas (NHL) and acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: Eligible patients had histologically proven relapse, age > or = 16 years, normal renal function, neutrophils > 500/microliter, platelets > 50,000/microliter, and no HIV infection, central nervous system disease, or serious neuropathy. Patients were treated with 2.0 mg/m2 of liposomal vincristine i.v. over 60 minutes q 14 days. Responders received up to 12 injections. RESULTS: Of the 51 registered patients, 35 are currently evaluable for response. Median age was 62 years (range 19-86), and 21 were male. The median number of prior regimens was 3 (range 1-10) and had included vincristine in all patients, of whom 51% were refractory to their last regimen. Serum LDH was high in 46%, and beta 2-microglobulin > 3.0 mg/l in 63% of patients. Of the 155 administered injections, 138 (89%) were at the 2.0 mg/m2 level. The median injected dose was 3.8 mg (range 2.6-4.8 mg), and median number of injections was 4 (range 1-12). Responses were seen in 14 of 34 (41%) patients with NHL (95% confidence intervals (95% CI) 25%-59%). Response rates were 10% for indolent, 71% for transformed, and 47% for aggressive NHL, but the 95% confidence intervals overlapped. Median progression-free survival was 5.5 months for responders. Grade 3-4 motor or sensory neuropathy was seen in 11, and caused termination of therapy in five patients. All five had prior neuropathy, two had previously received paclitaxel, one platinum, and two paclitaxel and platinum. Fever was detected in three patients, but there were no toxic deaths. CONCLUSIONS: Liposomal vincristine is active and well tolerated in this heavily pretreated population with relapsed NHL, but can be neurotoxic in a fraction of patients heavily exposed to prior neurotoxic agents. These data, if confirmed, would suggest a potential role for liposomal vincristine in the combination therapy of previously untreated patients with NHL. PMID- 10690393 TI - Paclitaxel plus vinorelbine: an active regimen in metastatic breast cancer patients with prior anthracycline exposure. AB - PURPOSE: To evaluate the anti-tumour activity and tolerance of the combination of paclitaxel plus vinorelbine in metastatic breast cancer (MBC) patients previously treated with anthracyclines. PATIENTS AND METHODS: Fifty-six MBC patients who have had at least one previous anthracycline-containing chemotherapy regimen were enrolled in this phase II trial. Patients received paclitaxel (135 mg/m2 over one hour infusion) and vinorelbine (30 mg/m2) both on day 1 of each three-week course of therapy (maximum eight courses or until disease progression was evident). RESULTS: Six complete and nineteen partial responses were observed among the fifty-four assessable patients (response rate of 46%, 95% CI: 33%-60%). Responses were observed in all disease sites and in all subsets of patients. The response rates when paclitaxel plus vinorelbine were used as first, second and third-line chemotherapy for metastases were 67%, 41% and 35%, respectively. The response rate among anthracycline-refractory patients was 46% (6 of 13). Median time to progression in the overall patient group was 28 weeks. The main toxicities (CTC grade 2 or more) were alopecia, myelosuppression and peripheral neuropathy (85%, 46% and 19% of patients, respectively). Nine patients (17%) had neutropenic fever in fifteen of the three hundred twenty-eight courses administered (5%). CONCLUSIONS: The combination of paclitaxel and vinorelbine on day 1 every three weeks is active in MBC patients with prior anthracycline exposure. The regimen is safe, well tolerated and convenient for the patients. PMID- 10690394 TI - Epstein-Barr virus association in classical Hodgkin's disease provides survival advantage to patients and correlates with higher expression of proliferation markers in Reed-Sternberg cells. AB - BACKGROUND: Most Epstein-Barr virus (EBV) associated lymphoproliferative disorders have high proliferation indices. However, classical Hodgkin's disease (cHD) is heterogeneous, with respect to proliferation index of the Reed-Sternberg cell (RS cell), and EBV association. Hence, we investigated whether cHD with and without EBV-association differ with respect to the proliferation index of the RS cells. Further we investigated whether this would have a bearing on patients survival. PATIENTS AND METHODS: We investigated 110 cases of cHD for: a) EBV association by immunohistochemical demonstration of EBV-latent membrane protein-1 and EBV encoded nuclear RNA 1 by mRNA in situ hybridisation; b) Proliferating cell nuclear antigen (PCNA) expression in the RS cells. RESULTS: EBV association was noted in 86 of 110 cases (78%). Higher PCNA expression (P = 0.004) and younger age (P = 0.001) correlated independently with EBV association. The 10 year relapse free survival (RFS) of EBV+ and EBV- patients were 60% and 44%, respectively (P = 0.03). The 10 year overall survival (OS) of EBV+ and EBV- patients were 85% and 64%, respectively (P = 0.03). EBV association maintained its significant impact on RFS and OS within Cox proportional hazard model. CONCLUSIONS: Our study suggests that EBV is likely to confer a higher PCNA expression and also contribute towards maintaining the RS cells of cHD in cell cycle. Hence, RS cells in EBV associated cHD would be more responsive to chemotherapy and radiotherapy associated DNA damage. Thus, EBV-association provides survival advantage to cHD patients treated with standard chemotherapy and radiotherapy protocols. PMID- 10690395 TI - Parathyroid hormone-related protein in metastatic breast cancer induced hypercalcemia: a case report. PMID- 10690396 TI - Phase II study of the multitargeted antifolate LY231514 (ALIMTA, MTA, pemetrexed disodium) in patients with advanced pancreatic cancer. AB - PURPOSE: To determine the safety and activity of LY231514 (ALIMTA, MTA, pemetrexed disodium, Eli Lilly and Co., Indianapolis, IN) in chemotherapy-naive patients with advanced pancreatic cancer. PATIENTS AND METHODS: Patients with unresectable or metastatic pancreatic cancer received LY231514 600 mg/m2 as a 10 minute infusion every three weeks. RESULTS: Forty-two patients were enrolled in this phase II trial. The median age was 60.3 (range 37-77) years; 79% had metastatic disease. Neutropenia was common (40% of patients > or = grade 3) but infectious complications were rare. Significant anemia or thrombocytopenia occurred in < 20% of patients. Non-hematologic toxicities included grade 2 or 3 skin reaction which was ameliorated by dexamethasone. Elevations of bilirubin or transaminases were infrequent (< 25% of patients) and did not require dose reductions or treatment delays. Thirty-five patients received two cycles of therapy and were evaluable for response. One complete (duration 16.2 months) and one partial (duration 6.9 months) were observed resulting in an objective response rate of 5.7% for evaluable patients. In addition, 17 patients (40%) had stable disease that lasted > or = 6 months in 5 patients. The median survival was 6.5 months, with 28% of patients alive at one year. CONCLUSIONS: LY231514 is a well-tolerated agent with minimal objective antitumor activity in pancreatic cancer. The median and one year survival times, which may be important indicators in phase II trials of new agents, are of interest. Combination trials of LY231514 in pancreatic cancer are planned. PMID- 10690397 TI - Socio-economic deprivation and stage of disease at presentation in women with breast cancer. AB - BACKGROUND: This study describes and compares the pathological prognostic factors and surgeon assessment of stage of breast cancer of women living in affluent and deprived areas to assess whether clinical stage at presentation may explain the known poorer survival outcomes for deprived women. PATIENTS AND METHODS: A population-based review of the case records of 417 women with breast cancer was carried out. RESULTS: No difference in pathological criteria was found between the 88% of women living in affluent and deprived areas for whom such data were available. Clinical assessment of the remaining 50 cases showed that women living in deprived areas were more likely to present with locally advanced or metastatic disease. CONCLUSION: The poorer survival of women from deprived areas with breast cancer may be explained by more deprived women presenting with advanced cancers. PMID- 10690398 TI - A phase I-II study of gemcitabine and paclitaxel in advanced non-small-cell lung cancer patients. AB - Thirty patients with chemotherapy-naive advanced non-small-cell lung cancer (NSCLC) were given escalating doses of paclitaxel (150, 175, 200 mg/m2) on day 1 in three consecutive cycles, together with a fixed dose of gemcitabine 1000 mg/m2 on days 1 and 8; cycles were repeated every three weeks. The dose escalation of paclitaxel was feasible in the majority of patients. Subsequently, 30 other NSCLC patients received a dose of 200 mg/m2 paclitaxel with gemcitabine 1000 mg/m2 in a phase II study. The major side effect was mild myelosuppression. A response rate of 24% was achieved in 49 fully evaluable patients. This regimen proved to be safe and easy to administer on an out-patient setting, and constitutes now one of the arms of the current EORTC randomized study for advanced NSCLC. PMID- 10690399 TI - A phase II study of pegylated liposomal doxorubicin for treatment of advanced hepatocellular carcinoma. AB - BACKGROUND: Pegylated liposomal doxorubicin has an enhanced efficacy and reduced toxicity compared with free doxorubicin. The efficacy and toxicity of pegylated liposomal doxorubicin was investigated in patients with hepatocellular carcinoma. PATIENTS AND METHODS: Patients with histologically confirmed, locally advanced or metastatic hepatocellular carcinoma and a Karnofsky index > 60% were included in this prospective single-arm study. Exclusion criteria were liver cirrhosis stage Child-Pugh C, previous chemotherapy, or chemoembolization. Pegylated liposomal doxorubicin was given in a dose of 30 mg/m2 every three weeks until progression of disease. After inclusion of five patients the dose could be escalated to 40 mg/m2 in absence of toxicity grade 3 and 4. RESULTS: Sixteen patients were evaluable for response. No objective response was achieved. The median survival time was 140 days (95% confidence interval: 126-154 days). Treatment toxicities grade > or = 3 comprised increased liver enzymes in patients with preexisting grade 1 or 2 elevation (n = 6), hematologic toxicity (n = 5), and hypersensitivity (n = 2). CONCLUSIONS: Pegylated liposomal doxorubicin is not effective for treatment of advanced hepatocellular carcinoma. The favorable toxicity profile was confirmed even in patients with underlying liver disease. PMID- 10690400 TI - Combination chemotherapy with irinotecan and adriamycin for refractory and relapsed non-Hodgkin's lymphoma. AB - Twenty-five patients with relapsed or refractory non-Hodgkin's lymphoma were treated by combination chemotherapy with irinotecan hydrochloride (CPT-11) and adriamycin (ADM): CPT-11, 25 mg/m2 on days 1 and 2; ADM, 40 mg/m2 on day 3. Nine (36%) of twenty-five patients achieved CR. Fairly good responses were seen in relapsed B-cell lymphomas (4 of 8 in diffuse large B-cell lymphoma and 2 of 2 in follicular lymphoma grade 1), and substantial responses in T-cell lymphomas (1 of 4 in peripheral T-cell lymphoma and 2 of 7 in adult T-cell leukemia/lymphoma). Leukopenia was frequent but tolerable, and diarrhea minimal. Combination chemotherapy with a reduced dose CPT-11 and ADM was useful in the treatment of relapsed non-Hodgkin's lymphoma. PMID- 10690401 TI - Incidence of venous thromboembolism in breast cancer patients during chemotherapy with vinorelbine, cisplatin, 5-fluorouracil as continuous infusion (ViFuP regimen): is prophylaxis required? PMID- 10690402 TI - History of plant population genetics. AB - This review of plant population genetics focuses on the genetic foundations of the processes that have led to documentable improvements in cultivated plants since the earliest domestications took place perhaps 13,000 years ago. Nearly all human civilizations have depended heavily on inbreeding plants (particularly wheat, barley, soybeans and other inbreeding legumes), as well as outbreeding vegetatively propagated species (white potatoes, yams) as their dietary standbys. The principal exception is maize (corn), an annual seed-produced outbreeder in nature. It is noteworthy that maize joined wheat, rice, and barley as a truly major crop worldwide only after its conversion to self-pollination combined with hybridization between favorably interacting inbred lines increased yield of maize several-fold in the twentieth century. PMID- 10690403 TI - Fas ligand-induced apoptosis. AB - The immune response is regulated not only by cell proliferation and differentiation, but also by programmed cell death, or apoptosis. In response to various stimuli, death factors bind to their respective receptors and activate the apoptotic death program in target cells. A cascade of specific proteases termed caspases mediates the apoptotic process. The activated caspases cleave various cellular components, a process that leads to morphological changes of the cells and nuclei, as well as to degradation of the chromosomal DNA. Loss-of function mutations in the signaling molecules involved in apoptosis cause hyper proliferation of cells in mouse and human. In contrast, exaggeration of this death cascade causes the destruction of various tissues. PMID- 10690405 TI - Molecular genetics of human retinal disease. AB - The past decade has witnessed extraordinary progress in retinal disease gene identification, the analysis of animal and tissue culture models of disease processes, and the integration of this information with clinical observations and with retinal biochemistry and physiology. During this period over twenty retinal disease genes were identified and for many of these genes there are now significant insights into their role in disease. This review presents an overview of the basic and clinical biology of the retina, summarizes recent progress in understanding the molecular mechanisms of inherited retinal diseases, and offers an assessment of the role that genetics will play in the next phase of research in this area. PMID- 10690406 TI - Lentivirus replication and regulation. AB - Lentiviruses are associated with chronic diseases of the hematological and neurological systems in animals and man. In particular, human immunodeficiency virus type 1 (HIV-1) is the etiological agent of the global AIDS epidemic. The genomes of lentiviruses are complex, encoding a number of regulatory and accessory proteins not found in other retroviruses. This complexity is reflected in their replication cycle, which reveals intricate regulatory pathways and unique mechanisms for viral persistence. In this review, we highlight some of these unique features for HIV-1, with particular focus on the transcriptional and posttranscriptional control of gene expression. Although our understanding of the biology of HIV-1 is far from complete, the knowledge gained thus far has already led to novel strategies for both virus intervention and exploiting the lentiviruses for therapeutic applications. PMID- 10690404 TI - Mechanisms of stationary phase mutation: a decade of adaptive mutation. AB - A decade of research on adaptive mutation has revealed a plethora of mutagenic mechanisms that may be important in evolution. The DNA synthesis associated with recombination could be an important source of spontaneous mutation in cells that are not proliferating. The movement of insertion elements can be responsive to environmental conditions. Insertion elements not only activate and inactivate genes, they also provide sequence homology that allows large-scale genomic rearrangements. Some conjugative plasmids can recombine with their host's chromosome, and may acquire chromosomal genes that could then spread through the population and even to other species. Finally, a subpopulation of transient hypermutators could be a source of multiple variant alleles, providing a mechanism for rapid evolution under adverse conditions. PMID- 10690407 TI - Shufflons: multiple inversion systems and integrons. AB - Conservative site-specific recombination functions to create biological diversity in prokaryotes. Simple site-specific recombination systems consist of two recombination sites and a recombinase gene. The plasmid R64 shufflon contains seven recombination sites, which flank and separate four DNA segments. Site specific recombinations mediated by the product of the rci gene between any two inverted recombination sites result in the inversion of four DNA segments independently or in groups. The shufflon functions as a biological switch to select one of seven C-terminal segments of the PilV proteins, which is a minor component of R64 thin pilus. The shufflon determines the recipient specificity in liquid matings of plasmid R64. Other multiple inversion systems as well as integrons, which are multiple insertion systems, are also described in this review. PMID- 10690408 TI - Messenger RNA stability and its role in control of gene expression in bacteria and phages. AB - The stability of mRNA in prokaryotes depends on multiple factors and it has not yet been possible to describe the process of mRNA degradation in terms of a unique pathway. However, important advances have been made in the past 10 years with the characterization of the cis-acting RNA elements and the trans-acting cellular proteins that control mRNA decay. The trans-acting proteins are mainly four nucleases, two endo- (RNase E and RNase III) and two exonucleases (PNPase and RNase II), and poly(A) polymerase. RNase E and PNPase are found in a multienzyme complex called the degradosome. In addition to the host nucleases, phage T4 encodes a specific endonuclease called RegB. The cis-acting elements that protect mRNA from degradation are stable stem-loops at the 5' end of the transcript and terminators or REP sequences at their 3' end. The rate-limiting step in mRNA decay is usually an initial endonucleolytic cleavage that often occurs at the 5' extremity. This initial step is followed by directional 3' to 5' degradation by the two exonucleases. Several examples, reviewed here, indicate that mRNA degradation is an important step at which gene expression can be controlled. This regulation can be either global, as in the case of growth rate dependent control, or specific, in response to changes in the environmental conditions. PMID- 10690409 TI - Mechanisms of mRNA surveillance in eukaryotes. AB - A conserved mRNA degradation system, referred to as mRNA surveillance, exists in eukaryotic cells to degrade aberrant mRNAs. A defining aspect of aberrant transcripts is that the spatial relationship between the termination codon and specific downstream sequence information has been altered. A key, yet unknown, feature of the mRNA surveillance system is how this spatial relationship is assessed in individual transcripts. Two views have emerged to describe how discrimination between proper and improper termination might occur. In the first view, a surveillance complex assembles onto the mRNA after translation termination, and scans the mRNA in a 3' to 5' direction for a limited distance. If specific downstream sequence information is encountered during this scanning, then the surveillance complex targets the transcript for rapid decay. An alternate view suggests that the downstream sequence information influences how translation termination occurs. This view encompasses several ideas including: (a) The architecture of the mRNP can alter the rate of key steps in translation termination; (b) the discrimination between a proper and improper termination occurs via an internal, Upf1-dependent, timing mechanism; and (c) proper termination results in the restructuring of the mRNP to a form that promotes mRNA stability. This proposed model for mRNA surveillance is similar to other systems of kinetic proofreading that monitor the accuracy of other biogenic processes such as translation and spliceosome assembly. PMID- 10690410 TI - Ribosome synthesis in Saccharomyces cerevisiae. AB - The synthesis of ribosomes is one of the major metabolic pathways in all cells. In addition to around 75 individual ribosomal proteins and 4 ribosomal RNAs, synthesis of a functional eukaryotic ribosome requires a remarkable number of trans-acting factors. Here, we will discuss the recent, and often surprising, advances in our understanding of ribosome synthesis in the yeast Saccharomyces cerevisiae. These will underscore the unexpected complexity of eukaryotic ribosome synthesis. PMID- 10690411 TI - The French school of genetics: from physiological and population genetics to regulatory molecular genetics. AB - French genetics had unusual beginnings. There are clear indications that the French biological establishment resisted Mendelian genetics strenuously from about 1910 to 1940. From about 1930 to 1950 several unconventional research programs with a strongly physiological orientation paved the way for the full entrance of French biology into genetics after World War II. This review examines some salient features of this history to clarify the strengths, weaknesses, and distinctive features of French genetics until about 1965. We suggest that after that data French genetics slowly merged into the international mainstream as genetics has become a largely molecular discipline. PMID- 10690412 TI - Mitochondrial genome evolution and the origin of eukaryotes. AB - Recent results from ancestral (minimally derived) protists testify to the tremendous diversity of the mitochondrial genome in various eukaryotic lineages, but also reinforce the view that mitochondria, descendants of an endosymbiotic alpha-Proteobacterium, arose only once in evolution. The serial endosymbiosis theory, currently the most popular hypothesis to explain the origin of mitochondria, postulates the capture of an alpha-proteobacterial endosymbiont by a nucleus-containing eukaryotic host resembling extant amitochondriate protists. New sequence data have challenged this scenario, instead raising the possibility that the origin of the mitochondrion was coincident with, and contributed substantially to, the origin of the nuclear genome of the eukaryotic cell. Defining more precisely the alpha-proteobacterial ancestry of the mitochondrial genome, and the contribution of the endosymbiotic event to the nuclear genome, will be essential for a full understanding of the origin and evolution of the eukaryotic cell as a whole. PMID- 10690413 TI - Genetics of chemotaxis and thermotaxis in the nematode Caenorhabditis elegans. AB - Molecular genetic analysis of chemotaxis and theramotaxis in Caenorhabditis elegans has revealed the molecular bases of olfaction, taste, and thermosensation, which, in turn, has demonstrated that sensory signaling in C. elegans is very similar to that in vertebrates. A cyclic nucleotide-gated channel (TAX-2/TAX-4) that is highly homologous to the olfactory and photoreceptor channels in vertebrates is required for taste and thermosensation, in addition to olfaction. A cation channel (OSM-9) that is closely related to a capsaicin receptor channel is required for olfactory adaptation in one olfactory neuron and olfactory sensation in the other olfactory neuron. A novel G alpha protein (ODR 3) is essential for olfactory responses in all olfactory neurons and aversive responses in a polymodal sensory neuron. A G protein-coupled seven-transmembrane receptor (ODR-10) is the first olfactory receptor whose ligand was elucidated. Using chemotaxis and thermotaxis as behavioral paradigms, neural plasticity including learning and memory can be studied genetically in C. elegans. PMID- 10690414 TI - Bacterial cell division. AB - Formation of the bacterial division septum is catalyzed by a number of essential proteins that assemble into a ring structure at the future division site. Assembly of proteins into the cytokinetic ring appears to occur in a hierarchial order that is initiated by the FtsZ protein, a structural and functional analog of eukaryotic tubulins. Placement of the division site at its correct location in Escherichia coli requires a division inhibitor (MinC), that is responsible for preventing septation at unwanted sites near the cell poles, and a topological specificity protein (MinE), that forms a ring at midcell and protects the midcell site from the division inhibitor. However, the mechanism responsible for identifying the position of the midcell site or the polar sites used for spore septum formation is still unclear. Regulation of the division process and its coordination with other cell cycle events, such as chromosome replication, are poorly understood. However, a protein has been identified in Caulobacter (CtrA) that regulates both the initiation of chromosome regulation and the transcription of ftsZ, and that may play an important role in the coordination process. PMID- 10690415 TI - Toward an integrated genetic epidemiology of parasitic protozoa and other pathogens. AB - Due to the increase of human migrations, the appearance of emerging and reemerging endemies, growing antibiotic resistance, and climatic changes, infectious diseases most probably constitute the major challenge for medicine in the next century. The advent of molecular methods of pathogen characterization has considerably improved our knowledge of the epidemiology of these diseases. However, the use of concepts of evolutionary genetics for interpreting "molecular epidemiology" data remains limited, although the application of such methods would broaden considerably the scope of this field of research, and allow epidemiologic and taxonomic approaches to be ascertained on a much firmer basis. In turn, pathogens, hosts, and vectors provide fascinating models for basic research. The artificial character of the border between "basic" and "applied" research is especially apparent with regard to the "integrated genetic epidemiology of infectious diseases" concept. The goal of this chapter is to evaluate the respective impact, on the transmission and pathogenicity of infectious diseases, of the host's, the pathogen's, and the vector's (for vector borne diseases) genetic diversity, and the interactions between these three parameters (coevolution phenomena). PMID- 10690416 TI - Plant retrotransposons. AB - Retrotransposons are mobile genetic elements that transpose through reverse transcription of an RNA intermediate. Retrotransposons are ubiquitous in plants and play a major role in plant gene and genome evolution. In many cases, retrotransposons comprise over 50% of nuclear DNA content, a situation that can arise in just a few million years. Plant retrotransposons are structurally and functionally similar to the retrotransposons and retroviruses that are found in other eukaryotic organisms. However, there are important differences in the genomic organization of retrotransposons in plants compared to some other eukaryotes, including their often-high copy numbers, their extensively heterogeneous populations, and their chromosomal dispersion patterns. Recent studies are providing valuable insights into the mechanisms involved in regulating the expression and transposition of retrotransposons. This review describes the structure, genomic organization, expression, regulation, and evolution of retrotransposons, and discusses both their contributions to plant genome evolution and their use as genetic tools in plant biology. PMID- 10690417 TI - Mammalian DNA mismatch repair. AB - DNA mismatch repair (MMR) is one of multiple replication, repair, and recombination processes that are required to maintain genomic stability in prokaryotes and eukaryotes. In the wake of the discoveries that hereditary nonpolyposis colorectal cancer (HNPCC) and other human cancers are associated with mutations in MMR genes, intensive efforts are under way to elucidate the biochemical functions of mammalian MutS and MutL homologs, and the consequences of defects in these genes. Genetic studies in cultured mammalian cells and mice are proving to be instrumental in defining the relationship between the functions of MMR in mutation and tumor avoidance. Furthermore, these approaches have raised awareness that MMR homologs contribute to DNA damage surveillance, transcription coupled repair, and recombinogenic and meiotic processes. PMID- 10690418 TI - Family values in the age of genomics: comparative analyses of temperate bacteriophage HK022. AB - HK022 is a temperate coliphage related to phage lambda. Its chromosome has been completely sequenced, and several aspects of its life cycle have been intensively studied. In the overall arrangement, expression, and function of most of its genes, HK022 broadly resembles lambda and other members of the lambda family. Upon closer view, significant differences emerge. The differences reveal alternative strategies used by related phages to cope with similar problems and illuminate previously unknown regulatory and structural motifs. HK022 prophages protect lysogens from superinfection by producing a sequence-specific RNA binding protein that prematurely terminates nascent transcripts of infecting phage. It uses a novel RNA-based mechanism to antiterminate its own early transcription. The HK022 protein shell is strengthened by a complex pattern of covalent subunit interlinking to form a unitary structure that resembles chain-mail armour. Its integrase and repressor proteins are similar to those of lambda, but the differences provide insights into the evolution of biological specificity and the elements needed for construction of a stable genetic switch. PMID- 10690419 TI - Meiotic chromosomes: integrating structure and function. AB - Meiotic chromosomes have been studied for many years, in part because of the fundamental life processes they represent, but also because meiosis involves the formation of homolog pairs, a feature which greatly facilitates the study of chromosome behavior. The complex events involved in homolog juxtaposition necessitate prolongation of prophase, thus permitting resolution of events that are temporally compressed in the mitotic cycle. Furthermore, once homologs are paired, the chromosomes are connected by a specific structure: the synaptonemal complex. Finally, interaction of homologs includes recombination at the DNA level, which is intimately linked to structural features of the chromosomes. In consequence, recombination-related events report on diverse aspects of chromosome morphogenesis, notably relationships between sisters, development of axial structure, and variations in chromatin status. The current article reviews recent information on these topics in an historical context. This juxtaposition has suggested new relationships between structure and function. Additional issues were addressed in a previous chapter (551). PMID- 10690420 TI - [Pathogen microevolution in the course of an infectious process]. PMID- 10690421 TI - [Inhibition of the reproduction of strains of the herpes simplex virus type 1 with drug resistance]. AB - Inhibition of type-1 herpes simplex strains resistant to acyclovir, phosphonoacetic acid and their combination by combined use of three drugs with different mechanisms of action capable of suppressing reproduction of the acyclovir resistant strain was studied. The combinations used were the following: Ara-A + ribavirin + phosphonoformic acid, Xylo-A + ribavirin + phosphonoformic acid and Ph-ACH + Ara-A + ribavirin. The former two combinations had a synergistic action on the standard strain L2 whose drug susceptibility had not undergone changes as well as on the acyclovir resistant strain. As for the strain resistant to phosphonoacetic acid and to acyclovir + phosphonoacetic acid the effect was additive. Ph-ACH + Ara-A + ribavirin had a marked synergistic action on all the strains tested. PMID- 10690422 TI - [The effect of heat shock on the growth and mycelial morphology of Streptomyces chrysomallus]. AB - The effect of various conditions of heat shock (1 hour at 35, 38, 40, 42, 45 and 50 degrees C) on the growth and morphological features of Streptomyces chrysomallus, an organism producing actinomycin, was studied. A definite regularity in the mycelium morphological changes at high temperatures was observed. After the shock at 35 and 38 degrees C the biomass volume and morphological features of the streptomycete did not markedly differ from those in the control. The shock at 40 degrees C induced the growth inhibition with decreasing the biomass volume by 50 per cent and appearance of submerged spores. When the shock conditions were more rigid (42, 45 and 50 degrees C) the mycelium growth lacked. It is of interest that the temperature of 42 degrees C induced abundant formation of the spores. With further increasing of the temperature to 45 and 50 degrees C the spore formation was not so abundant. The changes in the growth and development of the streptomycete are discussed in relation to the molecular mechanism of the cell protection from temperature shock. PMID- 10690423 TI - [The in-vitro effect of the leukocytic cationic protein preparation Intercide on Escherichia coli]. AB - Intercide is a cationic protein with the molecular weight of 11.0-11.5 kD from human leukocytes. The in vitro effect of its different concentrations (0.6 to 1.8 mg/ml) on populations of Escherichia coli M17 and K12 and 120 E.coli isolates from various sources such as water, feces of healthy humans and patients with extraintestinal escherichiosis was studied. The experiments with the bacterial suspensions and broth cultures demonstrated that Intercide had an antibacterial action on both the stationary and growing cells. However, some strains of E.coli were resistant to the lethal effect of Intercide. It was observed for the first time that in a concentration of 1.8 mg/ml Intercide was able to stimulate the biomass growth of some E.coli strains in broth culture. The factor analysis showed that the Intercide stimulating effect was more often evident with respect to extraintestinal escherichiosis pathogens with high anti-Intercide and antilysozyme activities. PMID- 10690424 TI - [A comparative analysis of the properties of bacteria in the genus Salmonella isolated from children in Latvia]. AB - Since social and economic changes in Latvia in 1991 the incidence of salmonellosis dramatically increased: more than 500 pediatric cases are registered every year. Specification of the properties of the Salmonella strains isolated in Latvia was undertaken. The study demonstrated that acute salmonellosis in pediatric inpatients was mainly due to S.typhimurium (78 per cent) and only in 22 per cent of the patients it was due to S.enteritidis. All the S.typhimurium isolates showed high antibiotic resistance defined by specific extended spectrum beta-lactamase (CTX-M-5). PMID- 10690425 TI - [New natural immunosuppressants. A comparison by their mechanism of action with cyclosporin A]. PMID- 10690426 TI - [The current approaches to enhancing the efficacy of antitumor chemotherapy by individual treatment optimization and the selective decrease in the toxic side effects of cytostatics]. PMID- 10690427 TI - [A broad-spectrum antimicrobial preparation--lomefloxacin (Maxaquin): the results of 10 years of its use in the clinics of Russia]. PMID- 10690428 TI - Comparison of biochemical polymorphisms and short tandem repeat (STR) DNA markers for paternity testing in rhesus monkeys (Macaca mulatta). AB - Genetic markers are indispensable for molecular and statistical genetic research involving nonhuman primates. Genetic markers must be used to ascertain parentage and to confirm the accuracy of pedigrees based solely on housing or demographic records; otherwise, the results of pedigree, linkage, or quantitative genetic analyses may be unreliable. Until recently, most genetic markers used in nonhuman primates were plasma proteins or isozyme polymorphisms, which were required in large numbers, because levels of genetic variation revealed by these markers were rather low. We compared the newer, PCR-amplified short tandem repeat markers (STRs) with a panel of classical biochemical polymorphic markers, for paternity determination among captive-bred rhesus monkeys. The STR markers exhibited an average genetic diversity of 64% and an expected paternity exclusion probability of 0.443. Both of these were greater than the average 54.5% genetic diversity and 0.298 exclusion probability exhibited by the biochemical markers. The STRs were much more efficient than the biochemical markers for parentage determination, since they required only half the amount of genetic typing data to resolve an average paternity case. Thus, the results of applying these two classes of genetic markers in paternity tests were somewhat different than expected on the basis of theoretical exclusion probabilities. These differences were probably due to inbreeding and other genetic differences among breeding colonies. Because they are more informative and provide rapid and efficient genetic data, STRs are now the method of choice for parentage determination and pedigree corroboration among nonhuman primates. PMID- 10690429 TI - Mitochondrial genetic variation in Chinese pigs and wild boars. AB - The mitochondrial DNAs (mtDNAs) from 30 pig breeds (29 Chinese native breeds and 1 European breed) and wild boars were investigated for restriction fragment length polymorphisms (RFLPs) to determine the phylogenetic relationships and genetic diversity among pig breeds and wild boars. Of the 24 enzymes used, 8 (AvaI, BclI, BglII, EcoRI, EcoRV, ScaI, StuI, and XbaI) detected polymorphisms. By combining the cleavage patterns for each enzyme, 108 individuals were sorted into eight mtDNA mitotypes. There are two haplotype lineages in domestic pigs, i.e., Chinese and European lineages. The pairwise nucleotide sequence divergence was calculated to be 0.56% between Chinese pigs and European pigs, suggesting that they might have diverged from a common ancestor approximately 280,000 years ago. The wild boars showed more extensive genetic variation, four mitotypes were detected in six wild boars. In addition, one of the Zhejiang wild boars was found to share the same mitotype with Chinese native pigs. A UPGMA tree based on genetic distance among mitotypes indicated that mtDNAs of Chinese pigs and European pigs are clearly divided into two clusters, and Chinese wild boars are more closely related to the Chinese pigs. Our results provide molecular evidence to support the previous hypothesis that pigs may be derived from two maternal origins, Asian and European wild boars. Chinese native pig breeds may have a single origin. PMID- 10690430 TI - Is the difference in alpha-amylase activity in the strains of Drosophila melanogaster with different allozymes due to transcriptional or posttranscriptional control? AB - The alpha-amylase in Drosophila melanogaster is a highly polymorphic enzyme, at both the allozyme level and the specific activity level. This enzyme changes its specific activity drastically depending on both food conditions and developmental stages, and it has been suggested that the ability to change its activity depending on the source of food has positive correlation with fitness. But the cause of the difference of specific activity among strains and food compositions is not known. In order to investigate the cause of these differences, we measured both the specific activity of amylase and the relative amount of Amy mRNA in eight strains of D. melanogaster with different electromorphs, in two food environments and two developmental stages. We found the following. First, the food-dependent activity change is regulated at the transcription level. Second, there was a significant correlation between specific activity and mRNA level among strains. So 80 to 90% of the specific activity difference can be explained by differences in the level of mRNAs, but the remaining part cannot. Finally, there were significant differences in specific activity per mRNA both among strains and between developmental stages. This suggests that there are differences in the catalytic efficiency of each allozyme, strain- or stage specific translation rate, enzyme stability, or differential use of two Amy loci. PMID- 10690431 TI - Apolipoprotein E polymorphism and plasma lipid levels in Native Mongolian sheep. AB - Apolipoprotein E (apoE) phenotypes were determined in 199 unrelated native sheep (Khalkhas line) of Central Mongolia, using a polyacrylamide gel isoelectric focusing-immunoblotting technique, and the plasma lipid levels in different phenotypes were assayed enzymatically. Twenty-eight phenotypes were identified in this sheep. In addition to all the previously detected seven apoE variants composing the phenotypes, four new variants were discovered, which were called E8, E9, E10, and E11. From the population data, these were found to be genetically controlled by four codominant alleles, designated APOE8, APOE9, APOE10, and APOE11, based on the same mode of inheritance as in the seven variants. These alleles were detected at a low frequency, in the range of 0.005 to 0.0126. The Khalkhas sheep differed most significantly from the Baruwal and Lampuchhre sheep of Nepal and the Vietnamese sheep with respect to the allele frequencies found in some Asian local sheep previously examined. Type 1/1 and/or 2/7 sheep had significantly higher plasma levels of total cholesterol and low density lipoprotein cholesterol than type 7/7 sheep (P < 0.05 and/or P < 0.02). PMID- 10690432 TI - Inheritance of isozyme variants in seed tissues of Pinus merkusii Jungh. & De Vriese. PMID- 10690433 TI - Career directions. Real-world guidance from successful biomedical professionals. AB - Talking to employers and successful biomedical professionals reveals a profile of traits in common. Successful biomedical professionals exhibit a desire to excel and a willingness to work hard. They push beyond personal comfort zones to identify weaknesses and build skills that may not come easily. Whether on the job or in the classroom, they are always learning. Short-term sacrifices are endured for the long-term payoffs provided by academic degrees and certification. Successful biomedical professionals challenge assumptions, especially their own, in order to advance their careers. When it comes to important career decisions, they solicit advice and do research. Ultimately, however, the decisions they make must be in accord with their own internal guidance systems. Career management is important to get right. Aside from the financial security and emotional satisfaction provided by career choices, there is the practical matter that, in a sense, the journey lasts a lifetime. PMID- 10690434 TI - Impact of CDMA wireless phone power output and puncture rate on hearing aid interference levels. AB - Interference between digital wireless phones and hearing aids occurs when the radiofrequency bursts from the phone transmission are demodulated by the hearing aid amplifier. The amplified interference signal is heard as a "buzz" or "static" by the hearing aid wearer. Most research and standards development activity has focused on worst-case scenarios with the phone operating at its maximum power. Since this power level is often not typical in urban and suburban settings, it is of value to determine the impact of lower power levels on the overall level of audible interference. Using a frequency analyzer, and several hearings aids and code division multiple access (CDMA) phones, the audio frequency spectrum of interference was recorded for each phone-aid combination and for a range of power levels producing from no interference to maximum interference. As phone power is increased, the interference signal becomes distinguishable from the ambient noise level and a linear response region is observed in which a specified increase in power output results in a proportional increase in the overall input referenced interference level (OIRIL). As power is increased beyond the linear region, the hearing aid enters a saturation region where an additional power increase results in a reduction or no increase in the OIRIL. The numeric differences in interference documented in this study were used in conjunction with the results of a previous study by the authors to determine the impact of reduced power on speech intelligibility and annoyance. The amount of improvement for a given power reduction depends on the radiofrequency immunity of the hearing aid and is substantial for hearing aids with poor immunity. For high-immunity aids, the level of audible interference remains low even at high phone power levels. PMID- 10690435 TI - Evolution of the optimum bidirectional (+/- biphasic) wave for defibrillation. AB - Introduction of the asymmetric bidirectional (+/- biphasic) current waveform has made it possible to achieve ventricular defibrillation with less energy and current than are needed with a unidirectional (monophasic) waveform. The symmetrical bidirectional (sinusoidal) waveform was used for the first human heart defibrillation. Subsequent studies employed the underdamped and overdamped sine waves, then the trapezoidal (monophasic) wave. Studies were then undertaken to investigate the benefit of adding a second identical and inverted wave; little success rewarded these efforts until it was discovered that the second inverted wave needed to be much less in amplitude to lower the threshold for defibrillation. However, there is no physiologic theory that explains the mechanism of action of the bidirectional wave, nor does any theory predict the optimum amplitude and time dimensions for the second inverted wave. The authors analyze the research that shows that the threshold defibrillation energy is lowest when the charge in the second, inverted phase is slightly more than a third of that in the first phase. An ion-flux, spatial-K+ summation hypothesis is presented that shows the effect on myocardial cells of adding the second inverted current pulse. PMID- 10690436 TI - A software tool for fetal blood flow analysis. AB - Doppler ultrasonography is a widely used technique for determination of the fetal blood flow pattern. Determination of the waveform qualities was done manually, with considerable inter- and intraobserver variations. In order to limit the variations and the time-consuming data entry, a Fetal Blood Flow Analysis software program was developed to facilitate ease of determination of Doppler signals. This article describes the development and unique features of the software program, made specifically to meet the obstetric and gynecology department's needs. PMID- 10690437 TI - Leadership and personality types. PMID- 10690438 TI - Internet addressing schemes. PMID- 10690439 TI - Mutual recognition agreements and what they mean to industry and regulatory bodies. PMID- 10690440 TI - Exercise in cardiac rehabilitation. PMID- 10690441 TI - The stigmatisation and denial of mental illness in athletes. PMID- 10690442 TI - Exercise and the prevention of back pain disability. PMID- 10690443 TI - The social patterning of exercise behaviours: the role of personal and local resources. PMID- 10690444 TI - Exercise in preventing falls and fall related injuries in older people: a review of randomised controlled trials. AB - OBJECTIVE: To assess the effectiveness of exercise programmes in preventing falls (and/or lowering the risk of falls and fall related injuries) in older people. DESIGN: A review of controlled clinical trials designed with the aim of lowering the risk of falling and/or fall injuries through an exercise only intervention or an intervention that included an exercise component. MAIN OUTCOME MEASURES: Falls, fall related injuries, time between falls, costs, cost effectiveness. SUBJECTS: A total of 4933 men and women aged 60 years and older. RESULTS: Eleven trials meeting the criteria for inclusion were reviewed. Eight of these trials had separate exercise interventions, and three used interventions with an exercise programme component. Five trials showed a significant reduction in the rate of falls or the risk of falling in the intervention group. CONCLUSIONS: Exercise is effective in lowering falls risk in selected groups and should form part of falls prevention programmes. Lowering fall related injuries will reduce health care costs but there is little available information on the costs associated with programme replication or the cost effectiveness of exercise programmes aimed at preventing falls in older people. PMID- 10690445 TI - Effects of one year of resistance training on the relation between muscular strength and bone density in elderly women. AB - OBJECTIVES: There is a paucity of long term studies on exercise training in elderly women. The purpose of this study was to investigate the effects of one year of progressive resistance exercise (PRE) on dynamic muscular strength and the relations to bone mineral density (BMD) in elderly women. METHODS: Forty four healthy sedentary women (mean age 68.8 years) volunteered for this study and were randomly assigned to either an exercise group or a control group. The exercise group were involved in three one hour sessions a week for 52 weeks of supervised PRE to strengthen the large muscle groups of the body, while the control group were instructed to continue their normal lifestyle. The exercise circuit included three sets of eight repetitions at 75% of one repetition maximum focused on the large muscle groups. BMD was measured by dual energy x ray absoptiometry (Lunar DPX) at the lumbar spine and at three sites in the proximal femur. Other selected parameters of physical fitness were also measured. RESULTS: Statistical analyses (analysis of covariance) showed significant strength gains (p < 0.01) in bilateral bench press (> 29%), bilateral leg press (> 19%), and unilateral biceps curl (> 20%). No significant difference between groups was evident in body weight, grip strength, flexibility, waist to hip ratio, or the sum of eight skinfolds. Significant relations (p < 0.05) were recorded between dynamic leg strength and the BMD of the femoral neck, Ward's triangle, and the lumbar spine. CONCLUSIONS: Significant strength changes, after one year of PRE, were evident in elderly women, and the muscle increases may parallel changes in BMD; however, correlation coefficients were moderate. PMID- 10690446 TI - Neutrophil function response to aerobic and anaerobic exercise in female judoka and untrained subjects. AB - OBJECTIVES: Recent studies have indicated reduced immunity in trained athletes. AIM: To assess the effects of aerobic and anaerobic exercise on the phagocytic process in 18-26 year old trained female judoka (n = 8) and untrained controls (n = 7). METHODS: Each subject participated randomly in two different testing sessions (aerobic, 20 minutes of treadmill running at 70-80% of maximal heart rate; anaerobic, Wingate anaerobic test). Venous blood samples were drawn before, immediately after, and 24 hours after each session. RESULTS: There were no significant differences in basal values of net chemotaxis (chemotaxis--random migration), bactericidal activity, and superoxide anion release between the judoka and the untrained women. There was a significant decrease in net chemotaxis 24 hours after the aerobic exercise in both the judoka (from 64 (19) to 39 (13) cells/field, p < 0.02) and the untrained controls (from 60 (7) to 47 (12) cells/field, p < 0.05). Bactericidal activity and superoxide anion release did not change significantly after aerobic exercise in either group. There were no significant changes in net chemotaxis, bactericidal activity, and superoxide anion release after anaerobic exercise in either the judoka or untrained women. CONCLUSIONS: The decrease in net chemotaxis after aerobic, but not after anaerobic, exercise, suggests that net chemotaxis is affected by the combination of exercise intensity and duration, and not by the exercise intensity itself. Similar effects of both exercise sessions in the judoka and the untrained women suggest that training had no effect on neutrophil function response to aerobic and anaerobic exercises. PMID- 10690447 TI - Balance control, flexibility, and cardiorespiratory fitness among older Tai Chi practitioners. AB - BACKGROUND: Tai Chi Chuan (TTC) exercise has beneficial effects on the components of physical condition and can produce a substantial reduction in the risk of multiple falls. Previous studies have shown that short term TCC exercise did not improve the scores in the single leg stance test with eyes closed and the sit and reach test. There has apparently been no research into the effects of TCC on total body rotation flexibility and heart rate responses at rest and after a three minute step test. METHODS: In this cross sectional study, 28 male TCC practitioners with an average age of 67.5 years old and 13.2 years of TCC exercise experience were recruited to form the TCC group. Another 30 sedentary men aged 66.2 were selected to serve as the control group. Measurements included resting heart rate, left and right single leg stance with eyes closed, modified sit and reach test, total body rotation test (left and right), and a three minute step test. RESULTS: Compared with the sedentary group, the TCC group had significantly better scores in resting heart rate, three minute step test heart rate, modified sit and reach, total body rotation test on both right and left side (p < 0.01), and both right and left leg standing with eyes closed (p < 0.05). According to the American Fitness Standards, the TCC group attained the 90th percentile rank for sit and reach and total body rotation test, right and left. CONCLUSION: Long term regular TCC exercise has favourable effects on the promotion of balance control, flexibility, and cardiovascular fitness in older adults. PMID- 10690448 TI - A comparison of lactate concentration in plasma collected from the toe, ear, and fingertip after a simulated rowing exercise. AB - OBJECTIVE: To examine the validity of using blood taken from the toe for the assessment of plasma lactate concentration in rowers. To achieve this, values were compared with those taken from the fingertip and earlobe. METHODS: Nine subjects exercised at two separate submaximum workloads on the Concept II rowing ergometer. The loads, each lasting four minutes, elicited mean (SD) heart rate responses of 160.1 (8.5) and 180.1 (5.7) beats/min, which corresponded to 76.4 (6.1)% and 91.9 (4.7)% of the estimated heart rate maximum of the subjects. Blood was simultaneously removed after the cessation of exercise by three experimenters and was analysed for plasma lactate concentration. RESULTS: At 76.4% of estimated heart rate maximum, the mean (SD) plasma lactate concentrations sampled from the fingertip, toe, and earlobe were 6.36 (1.58), 5.81 (1.11), and 5.29 (1.24) mmol/l respectively. At 91.9% of estimated heart rate maximum, respective values were 8.81 (2.30), 8.53 (1.37), and 8.41 (2.35) mmol/l. No significant differences (p > 0.05) were found between any of the sites at either work intensity. CONCLUSIONS: The toe may offer a practical alternative for assessing the concentration of lactate during rowing, having the advantage that repeated blood samples can be removed without interruption of the rowing action. PMID- 10690449 TI - Isokinetic performance and shoulder mobility in elite volleyball athletes from the United Kingdom. AB - OBJECTIVES: To evaluate the differences in strength and mobility of shoulder rotator muscles in the dominant and non-dominant shoulders of elite volleyball players. METHODS: Isokinetic muscle strength tests were performed at speeds of 60 and 120 degrees/s, and shoulder mobility was examined in ten players from the England national men's volleyball squad. The subjects also completed a questionnaire that included a visual prompt and analogue pain scale. RESULTS: The range of motion of internal rotation on the dominant side was less than that on the non-dominant side (p < 0.01). The average peak strength at 60 degrees/s external eccentric contraction was lower than that of internal concentric contraction in the dominant arm, but was higher in the non-dominant arm. Six of the ten subjects reported a shoulder problem, described as a diffuse pain located laterally on the dominant shoulder. CONCLUSIONS: These elite volleyball players had a lower range of motion (internal rotation) and relative muscle imbalance in the dominant compared with the non-dominant shoulder. PMID- 10690450 TI - Factors associated with hip joint rotation in former elite athletes. AB - OBJECTIVES: To study factors associated with passive hip rotation range of motion (ROM) in former elite male athletes. METHODS: Athletes were interviewed about hip pain, disability, lifetime occupational loading, and athletic training. The passive hip rotation was measured with a Myrin inclinometer in 117 former elite male long distance runners, soccer players, weight lifters, and shooters aged 45 68 years. Magnetic resonance imaging was used to detect hip osteoarthritis. RESULTS: There were no differences in passive hip rotation ROM between the four athlete groups nor between diverging lifetime loading patterns associated with occupational or athletic activities. Among the subjects without hip osteoarthritis, hip pain, and hip disability according to a stepwise linear regression analysis, the only factor that was associated with the passive hip rotation ROM was body mass index (BMI), explaining about 21% of its variation. Subjects with high BMI had lower passive hip rotation ROM than those with low BMI. There was no right-left difference in the mean passive hip rotation ROM in subjects either with or without hip osteoarthritis as determined by magnetic resonance imaging. Nevertheless, hip rotation ROM was clearly reduced in a few hips with severe caput deformity. CONCLUSIONS: Long term loading appears to have no association with passive hip rotation ROM. On the other hand, the hip rotation value was lower in subjects with high BMI than in those with low BMI. A clear right-left difference in hip rotation was found only in those subjects who, according to our magnetic resonance imaging criteria, had severe hip osteoarthritis. These findings should be taken into account when hip rotation ROM is used in the clinical assessment of hip joints. PMID- 10690451 TI - Correlation of bone scintigraphy and histological findings in medial tibial syndrome. AB - OBJECTIVE: To correlate bone scintigraphy and histopathological findings in patients with medial tibial syndrome. METHODS: Twenty patients (32 limbs) with a clinical diagnosis of medial tibial syndrome had surgery. Bone scintigraphy before the operation was compared with the histological appearance of bone and periosteal specimens obtained at surgery. RESULTS: Delayed bone scintigraphy showed normal appearance in 11 limbs, characteristic diffuse tubular pattern uptake in 16 limbs, and focal uptake in five. Periosteal histology disclosed fibrous thickening as the most common finding associated with increased vascularity, occasionally with chronic inflammatory cell infiltration, haemosiderin, and acid mucopolysaccharide deposition. Loss of osteocytes was the main finding of bone histology associated with some enlargement of lacunae and lamellar structure disruption. A grading system was used to score normal and abnormal histological appearance. For analysis the findings were regrouped to provide tables using Fisher's exact test. There was no correlation between bone scintigraphy and the histology of bone and periosteum, but two interesting observations were noted. Those cases with periosteal thickening had mostly normal bone scan appearance (p = 0.0028). Those cases with low levels of osteocyte loss had mostly abnormal bone scintigraphy. CONCLUSION: Abnormal histological appearance of bone and periosteum is a feature of medial tibial syndrome. These histological findings show poor correlation with bone scintigraphy. The exact pathogenesis of this syndrome remains unclear. PMID- 10690452 TI - Sport related proximal femoral fractures: a retrospective review of 31 cases treated in an eight year period. AB - In an eight year period, 31 patients with proximal femoral fractures resulting from sports accidents were treated by implantation of either a Gamma nail or a dynamic hip screw. Return to work or sports and the time to bone healing did not differ very much between the treatments. Gamma nailing was clearly the best with regard to stability and time to full mobilisation (4.5 days), but required 39 minutes to perform compared with insertion of a dynamic hip screw (27 minutes). The incidence of complications and malalignments did not differ very much between the two, although, when Gamma nailing was first used in the authors' clinic, more intraoperative complications occurred than with the dynamic hip screw. Stable pertrochanteric fractures may be treated with a dynamic hip screw. Unstable pertrochanteric or subtrochanteric fractures are treated with a Gamma nail at the authors' institution. PMID- 10690453 TI - Stages in the development of a research project: putting the idea together. AB - We have considered some of the most important factors involved in designing a viable study that will adequately address the research question. Although we do not profess to be experts in all aspects of the above, we have learned through experience that attention to many of the above points will help to avoid frustration during the experimental process and when the study is presented for external review and subsequent presentation and publication. Good luck in your research. PMID- 10690454 TI - Giant retinal tears resulting from eye gouging in rugby football. AB - A 29 year old myopic man sustained two separate giant retinal tears in his right eye following deliberate eye gouging during a rugby tackle. These were successfully repaired by vitrectomy and intraocular silicone oil injection. Although the postoperative course was complicated by pupil block glaucoma, he regained corrected visual acuity of 6/5 after oil removal. This injury highlights the potentially sight threatening nature of this type of rugby injury and the importance of early referral for specialist treatment. PMID- 10690455 TI - Redefining the overtraining syndrome as the unexplained underperformance syndrome. PMID- 10690456 TI - Upper airways obstruction. PMID- 10690457 TI - Use of imaging data for predicting clinical outcome. PMID- 10690458 TI - What is sports medicine? Medical students don't know. PMID- 10690459 TI - [Gertrude Belle Elion (1918-1999): brilliant chemist, discoverer of modern antivirals and Nobel Prize for medicine]. PMID- 10690460 TI - [Bacterial meningitis in the adult. Study of 85 cases observed in the infectious disease unit of the Fondation Jeanne Ebori (F.J.E.), Libreville, Gabon]. AB - We conducted a retrospective review to specify the frequency, identify the aetiological factors of bacterial meningitis in adults (BMA) and to evaluate the therapeutic protocol used. This study was conducted on 85 (BMA) cases of hospitalised patients between January 1991 and December 1995 (5 years) on our service. The BMA represented 3% of all admissions for infectious diseases at the Foundation Jeanne Ebori in Libreville. It occurred in an endemosporadic fashion. All patients were Black Africans with an average age of 33 years (range: 16-60 years). Males predominated by a ratio of 2.4. Tha patients were seen late in the evolution of the disease, as shown by the folloxing clinical signs: neuropsychic problems (100%), 25 patients (29%) were in a profound coma, 5 (6%) had a hemiplegia, 2 (2%) an hypoacousie and 1 (1%) seizure. Aetiological factors were found in 17 cases (20%) to be in the ORL sphere (sinusitis: n = 8, ear infection: n = 4), pneumopathies (n = 4) and one case of breach dure-mere. The predominant germ was pneumocoque, isolated in 55 cases (65%), 15 cases had a LCR clear (18%). Bacteria gram negative (6%) were identified in the immunocompromised HIV. Third generation cephems had an efficiency higher than beta lactamines: 83% against 73%. The mortality was 18%; 3% of the remaining patients had neurological deafness. The seriousness of the results of this survey calls for the urgent implementation of a surveillance programme. PMID- 10690461 TI - [Genotypic diversity of Mycobacterium tuberculosis in the Guiana-Antilles region]. AB - This investigation dealt with 226 strains (1 isolate/patient) of Mycobacterium tuberculosis isolated in the French West Indies and French Guiana over a three year period (1994-1996). The genotypic diversity of the isolates was investigated using various molecular markers; essentially two PCR-based rapid methods, namely spoligotyping and double-repetitive-element (DRE)-PCR, as well as three restriction fragment length polymorphism (RFLP)-based methods, namely IS6110 RFLP, DR-RFLP and PGRS-RFLP. Out of 226 isolates investigated, a total of 166 isolates were distributed in 31 spoligotype-defined clusters containing 2-31 strains, which corresponded to a rate of 73% of primary clustering. After secondary typing with DRE-PCR, IS6110-RFLP, DR-RFLP and/or PGRS-RFLP, molecular clonality was established for 73 isolates organised in 25 clusters (32% of clustered isolates). Considering one reactivation case per cluster, the rate of recent transmission was estimated to a minimal rate of 21%, however the available epidemiologic information led to the positive conclusion for only 14% of cases. The data obtained demonstrated the presence of common genotypes of M. tuberculosis among the three overseas French territories, i.e. Guadeloupe, Martinique and French Guiana. The results obtained during this retrospective study clearly indicate the importance of future prospective epidemiological investigations around the clustered cases of tuberculosis, so as to detect the persisting foci of endemic disease and characterize the chain of transmission as well as the subpopulations which are at an increased risk of contracting and/or propagating the disease. Last but not least, the present study also deals with a first phylogenetic approach of M. tuberculosis based on a comparison of the spoligotyping results obtained locally with those reported elsewhere in the world. PMID- 10690462 TI - [Dientamoeba fragilis: pathogenic flagellate?]. AB - INTRODUCTION: D. fragilis is an intestinal protozoa whose pathogenic characteristics are increasingly recognized. The aim of this study is to specify the epidemiologic, biological and clinical aspects of this protozoa. MATERIAL USED AND METHODOLOGY: Survey conducted on 27,058 parasitological test of stools in parasitology-mycology laboratory of the Sfax University Hospital over a period of 5 years. RESULTS: 11,254 parasitological test of stools were positive (41.6%) of which 89.3% comprised protozoa. D. fragilis was found in 1497 cases (13.3% of the positive cases). In 65% of these cases, it was associated with other intestinal parasites in particular Blastocystis hominis (40.3%), Endolimax nanus (24%), Entamoeba coli (6%), Giardia intestinalis (5.7%) and Enterobius vermicularis (5%). Those patients having a parasitism with isolated D. fragilis were predominantly female and young subjects (< 20 years). Clinical signs included abdominal pain (88.5%), anorexia (50%), alternating diarrhoea and constipation (40.4%) and diarrhoea (21%) with mucus in 7.6%. DISCUSSION: D. fragilis is today classified in the group of flagellates and we share the opinion of the majority of the authors as to its real pathogenic capacity. PMID- 10690463 TI - [Ecology of Lymnaea truncatula Muller, intermediate host of Fasciola hepatica Linne in the microclimate of Tozeur (southeast of Tunisia)]. AB - Systematic visits were carried out in the traditional Tozeur' oasis (Tunisia), in 1997/1998 in order to study the ecology of Lymnaea truncatula as well as its density in relation to habitat characteristics in the hydrographical networks of the oases. Lymnaea truncatula was enumerated on 1 m2 surface by habitat, according to the quadrat method. Samples of soil, irrigation and draining water were also analysed. The density of Lymnaea truncatula detected in autumn was constantly higher. It depends on the nature of the habitat: 54.8% in the secondary irrigation canals, 34.8% in those of draining, 7.6% in the seguia and 2.9% in the principal draining canal. Ecological factors--permanent dampness, suitable luminosity, basic pH of soil and water, temperature between 10 and 28 degrees C--contribute to the multiplicity of snails. PMID- 10690464 TI - [Human dirofilariasis in Corsica: a new local case. Review of reported cases]. AB - A case of human dirofilariasis with Dirofilaria (Nochtiella) repens, located in a subcutaneous nodule of the arch of the foot is reported in a 64-year-old man living in the surroundings of Porto Vecchio in Corsica. The histological examination of this nodule has revealed a gravid female nematode, dead some weeks or months before its surgical excision. It is the second case of human dirofilariasis observed in a patient living on the island. The authors are surprised by the number of patients reported in medical literature (10 altogether) as having contracted the infection probably during a short stay in Corsica. This paper seeks to collect the fragmented available data on the existence in Corsica of nematodes in dogs and its possible vectors. PMID- 10690465 TI - [Malaria transmission in an area of future vaccination in equatorial forest of south Cameroon]. AB - In order to describe malaria transmission in a future antigamete vaccine trial area, a longitudinal entomological study was conducted, together with parasitological and immunological surveys, from June 1997 to May 1998 in two nearby villages in a tropical rain forest area 100 km east of Yaounde. Koundou is located along the main road in an open and degraded environment combining cultivated lands and forests; Ebolakounou is located 5 km from the road in forest surroundings. Indoor mosquito night catches no human volunteers were performed twice a month, in ten houses. We determined the entomological infection rate as 176 infected bites per human per year in Koundou (47.7% for An. moucheti, 47.3% for An. gambiae and 5% for An. funestus) and only 17.7 infected bites/human/year in Ebolakounou, with An. gambiae only. Transmission appears to be ten times higher in the village situated in a degraded environment than in the village situated in the rainforest. PMID- 10690466 TI - [Variability of in vitro activity of proguanil and cycloguanil on erythrocyte stages of Plasmodium falciparum as a function of culture conditions]. AB - The in vitro activity of proguanil, cycloguanil (active metabolite of proguanil), pyrimethamine, and chloroquine was determined for 14 isolates of Plasmodium falciparum and the chloroquine-resistant W2 clone. In vitro assays were performed by using different types of RPMI 1640 culture medium and incubation period. The use of the standard RPMI medium or RPMI medium containing low concentrations of folate and para-aminobenzoic acid increases the 50% inhibitory concentrations of cycloguanil and pyrimethamine, as compared with the use of folate- and para aminobenzoic acid-free RPMI medium. The concentrations of folate and para aminobenzoic acid did not affect the in vitro activity of proguanil and chloroquine. However, prolongation of the incubation period from 42 to 66 hours decreased the 50% inhibitory concentrations of all test compounds. The weak antagonism in vitro between chloroquine and proguanil or cycloguanil does not seem to have any repercussion on the in vivo efficacy of chloroquine-proguanil combination. PMID- 10690467 TI - [Chloroquine sensitivity of Plasmodium falciparum at the Gamkalley Clinic and the Nigerian armed forces PMI (Niamey, Niger)]. AB - In vivo tests for Plasmodium falciparum were carried out in 1998 during the rainy season among children in Niamey, in the Republic of Niger. Chloroquine was prescribed at 25 mg/kg for 3 days in febrile patients with uncomplicated P. falciparum malaria. Forty-five 1-5 year-olds and thirty-three 6-15 year-olds were included in the study. A group of 53 adult patients was also surveyed to evaluate the efficacy of chloroquine in semi-immune persons. Body temperature and blood smears including parasitemia were recorded on days 0, 3, 7 and 14. Less than 10% of the patients were delinquent. Around 75% of the patients were successfully treated in the 1-5 year-olds and 6-15 year old age groups. Relapses were observed in 20% of the 1-5 year-olds (early relapses 8.9%, late relapses 11.1%) and in 16.7% in the 1-15 year-olds (early relapses 6.4%, late relapses 10.3%). Among adults, successful treatment was obtained in 86.8% of the cases and early and late relapses were respectively observed in 3.8% and 1.9% of the cases. All the patients with malaria relapses were cured with second-line treatments (pyrimethamine-sulfadoxine or quinine). According to these results, chloroquine resistance appears to be moderate in Niamey. Therefore chloroquine should remain the first line treatment of uncomplicated P. falciparum malaria in this population. PMID- 10690468 TI - [Sleeping sickness in children at Bobo-Dioulasso Hospital Center: apropos of 3 cases]. AB - The authors report three observations of trypanosomiasis in children aged 3 to 13 years from Ivory Coast and Burkina Faso. Two cases were imported from Cote d'Ivoire and one originated from an old endemic area of Bobo-Dioulasso region in Burkina Faso. Clinical features were comparable to classical descriptions in adults but neurological findings were dominant. Two children were at the lymphatic stage. Treatment with melarsoprol in two cases and eflornithine in one case led to complete recovery. Active epidemiologic surveillance of this zoonosis should be maintained and the devastating pandemic of the beginning of the century should be remembered. PMID- 10690469 TI - [Public hospitals in Sub-Saharan African countries and their perspectives]. AB - Hospitals have a very important role to play within the care system of Sub Saharan African countries, not only because they care for patients sent by other health services, but also because they participate in the training of health professionals. In spite of many reforms, they are trapped in a vicious circle, which, to be broken, means moving from too highly stratified administrative system to a system of enterprise. This system must focus on performance, while guaranteeing necessary public service. Such change requires adopting new methods and ways of thinking for providing a complete health service for the patients, that is adequate access to quality health care. This will entail hospitals being given a free hand in their management, effective measurements of quality and expenditure evaluation being devised and supervision by the state Health department. By allowing hospitals to attract more affluent members of society, they will be better able to meet their obligations towards the poor. PMID- 10690470 TI - [University and tropical medicine]. AB - Two "tropical medicine" coexist: a "clinical tropical medicine" concerning imported tropical diseases in North countries who is worked at clinical departments and travellers clinics and a multidisciplinary "international health", concerning tropical countries development, who is the concern of South and North Institutes cooperation. International health expansion compel to collaborate with foreign universities and institutes in and beyond the French speaking area. The scattering of practitioners, teachers and research workers in the midst of various French institutions and distance from field limit the academic policy of development. Harmonizing national and European tropical medicine teachings belong to inter-academic gathering. Programming and financing of international health come under an inter-ministerial agency. PMID- 10690471 TI - [Reemergence of yellow fever in West Africa: lessons from the past, advocacy for a control program]. AB - In French speaking West Africa, yellow fever vaccine became compulsory in 1941 for the entire African and European population. From 1941 to 1960, 146 million doses were distributed and the number of yellow fever cases declined sharply. No case was reported from 1954 to 1960. As a result of an interruption in systematic immunization after 1960, ten major epidemics broke out in West Africa between 1965 and 1995 (over 200,000 cases and 40,000 deaths). In 1967, the WHO programme for eradication of smallpox was initiated and it mobilized WHO's energy and finances. The expanded programme of immunization (EPI) was initiated in 1977 but it did not include the yellow fever vaccine. In 1978, Primary Health Care advocated an immunization strategy through fixed health facilities. In 1986, to amend this strategy, WHO recommended accelerating EPI progress and instituting National Immunization Days (NIDs). In 1990, a recommendation was made to include the yellow fever vaccine in the EPI. In 1997, the target of global poliomyelitis eradication by the year 2000 reinforced the NID programme and led to the use of mobile teams. At a time when a measles eradication programme is going to take over from the poliomyelitis programme, we must firmly advocate not omitting the yellow fever vaccine as was the case in 1977. Indeed, in yellow fever endemic areas, WHO recommends a simultaneous association of yellow fever and measles vaccines for nine month-old infants. This opportunity must be seized to initiate a yellow fever control programme. PMID- 10690472 TI - [Protective effect of clothing impregnated with permethrin against D. reticulatus and D. marginatus in an open biotope of central western France]. AB - During the period of major tick activity, in April and May 1998, in the Saint Maixent l'Ecole area of Poitou, a comparative study was carried out in order to evaluate the protective effect of garments impregnated with permethrin cis/trans 25/75. Three groups of soldiers made up respectively of 208, 218 and 427 men were involved for 2 or 3 days in this experiment. Respectively 107, 107 and 215 wore impregnated uniforms. Out of a total of 319 ticks, 3 were lxodes ricinus, 305 Dermacentor marginatus and 11 D. reticulatus. There was a significant difference in both the intensity (number of ticks per individual, P < 0.0001) and prevalence (number of individuals with ticks, P < 0.001) of ticks on individuals between impregnated and non impregnated uniforms. The repellent effect of permethrin on ticks was observed at the site of preferential tick attachment (normally the head of the host for these two species of Dermacentor in France) where the number of ticks was significantly lower in impregnated uniforms. PMID- 10690473 TI - [Oral receptivity of Aedes aegypti formosus from Franceville (Gabon, central Africa) for type 2 dengue virus]. AB - Dengue is widely distributed in the tropics but epidemic activity was rarely reported in Africa before the 1980's. In the past 15 years, increased epidemic dengue fever has been reported both in East and West Africa, raising concern about the ability of local populations of Aedes aegypti to transmit dengue viruses. Ae. aegypti is present in two forms in Africa: Ae. aegypti aegypti and Ae. aegypti formosus. This latter form, much darker, was not originally a local species but is now colonizing artificial breeding sites within cities. We have been able to demonstrate the oral susceptibility for dengue type 2 virus of Ae. aegypti formosus collected in Franceville, Gabon (Central Africa). However, these mosquitoes sampled exhibited lower infection rates than those of a control colony of Ae. aegypti aegypti originating from French Polynesia. PMID- 10690474 TI - [Present status of an arbovirus infection: yellow fever, its natural history of hemorrhagic fever, Rift Valley fever]. AB - In the early 20th century, when it was discovered that the yellow fever virus was transmitted in its urban cycle by Aedes aegypti, measures of control were introduced leading to its disappearance. Progressive neglect of the disease, however, led to a new outbreak in 1927 during which the etiological agent was isolated; some years later a vaccine was discovered and yellow fever disappeared again. In the 1960s, rare cases of encephalitis were observed in young children after vaccination and the administration of the vaccine was forbidden for children under 10 years. Five years later, a new outbreak of yellow fever in Diourbel, Senegal, was linked to the presence of Aedes aegypti. In the late 1970s, the idea of a selvatic cycle for yellow fever arose. Thanks to new investigative techniques in Senegal and Cote d'Ivoire, the yellow fever virus was isolated from the reservoir of virus and vectors. The isolated virus was identified in monkeys and several vectors: Aedes furcifer, Aedes taylori, Aedes luteocephalus. Most importantly, the virus was isolated in male mosquitoes. Until recently, the only known cycle had been that of Haddow in East Africa. The virus circulate in the canopea between monkeys and Aedes africanus. These monkeys infect Aedes bromeliae when they come to eat in banana plantations. This cycle does not occur in West Africa. Vertical transmission is the main method of maintenance of the virus through the dry season. "Reservoirs of virus" are often mentioned in medical literature, monkeys having a short viremia whereas mosquitoes remain infected throughout their life cycle. In such a selvatic cycle, circulation can reach very high levels and no child would be able to escape an infecting bite and yet no clinical cases of yellow fever have been reported. The virulence--as it affects man--of the yellow fever virus in its wild cycle is very low. In areas where the virus can circulate in epidemic form, two types of circulation can be distinguished. Intermediate yellow fever--a term coined to define epidemia which do not correspond exactly to urban yellow fever. The cycle involves men and monkeys through wild vectors as Aedes furcifer but also through Aedes aegypti and the mortality rate is much lower than for urban epidemics. In urban yellow fever, man is the only vertebrate host involved in the circulation of the virus, the vector being generally Aedes aegypti. This vector maintains a selective pressure, increasing the transmission of virus capable of producing high viremia in man. In the selvatic cycles, two cycles can be distinguished: one of maintenance which does not increase the quantity of virus in circulation and one of amplification which does increase this quantity. As we shall see, it develops into an epizootic form but also in an epidemic form in man. When the decrease in yellow fevers across Africa is considered, it appears that all major epidemics occur in West Africa inspite of the presence of wild cycles of the yellow fever virus in Central and East Africa. For the rare epidemics that have occurred there, the vector has never been Aedes aegypti. In a recent outbreak in Kenya, the vector was Aedes bromeliae. The examination of part of the gene encoding for envelope protein showed the presence of two geographical types corresponding to West-Africa and Central East-Africa. Clinically speaking, yellow fever is an haemorrhagic fever with hepatitis similar to other haemorrhagic fevers such as Rift Valley fever. When, in 1987, an outbreak of haemorrhagic fever occurred in southern Mauritania, for several days it was thought to be yellow fever. Four days later, the diagnosis was corrected by isolating and identifying the virus as that of Rift Valley fever (RVFV). RVFV causes several pathogenic syndromes in human beings: acute febrile illness, haemorrhagic fever, haemorrhagic fever with hepatitis, nervous syndromes or ocular disease. Mortality rate was high for haemorrhagic fever with hepatitis, reaching 36%. (ABST PMID- 10690475 TI - [Poland: cholera to typhus, 1831-1950]. AB - In this article devoted to Poland's direct and indirect role in the elaboration of contemporary international health structures and to her reputation as an epidemic reservoir of Europe, we consider how Poland came to be perceived as the cordon sanitaire of the West. Traditionally seen as upholding Western values, in the 19th and 20th centuries the country became increasingly associated with "Eastern plagues"-cholera and then typhus-coming from Russia and which could spread to the rest of Europe if Poland did not manage to contain them. When Poland was reconstituted as a nation-state in 1918, the new country won international recognition through her successful attempts to contain a typhus epidemic sweeping westwards from Russia. The Polish government convened the first European, League sponsored, health conference following the First World War. A Polish doctor, L. RAJCHMAN, was chosen to head up the League of Nations Health Organisation (forerunner of the WHO) and later (1946) founded UNICEF. Finally, we examine the key issue of exanthematous typhus in both world wars, exemplifying how a disease can come to be "ideologized", in this case by Nazi Germany. Typhus was the pretext used- in the name of "public health"-for segregating Polish citizens of Jewish origin and even killing them. Paradoxically, typhus was in the process of being eradicated when the war began and German policy of mass resettlements, sequestration, and starvation only spurred the epidemic they supposedly wished to control. PMID- 10690476 TI - [Filariasis in Haiti: a century of history]. AB - Wuchereria bancrofti and Mansonella ozzardi are both endemic in Haiti. Over the last hundred years, these human parasites have been by turn investigated and disregarded. Between 1894 and 1914, Haitian physicians encouraged by Dr. Leon AUDAIN studied the clinical and biological impact of W. bancrofti in the numerous infested patients in Port-au-Prince. During the American occupation (1915-1934), the presence of M. ozzardi was recognized by a Rockefeller mission and a first investigation of filariasis distribution in the country was carried out. Between 1935 and 1971, interest in the parasites ceased. However, many studies of W. bancrofti and M. ozzardi and their vectors have been conducted from 1972 until today. Lymphatic filariasis remains a great health hazard in localized leeward foci, where climatic conditions favourise the survival of the vector Culex quinque-fasciatus. Urban foci have been remarkably stable for the last 70 years in northern Haiti and along the Gulf of the Gonave coast. Parasitological indices are high and the impact on public health is great. Ozzardiasis is prevalent in the rural coastal areas of northern and southern Haiti, where the principal vectors Culicoides furens and C. barbosai breed in abundance. The control of lymphatic filariasis is now possible and should be a public health priority in Haiti. PMID- 10690478 TI - Modern medicine. Too much of a good thing? PMID- 10690477 TI - [Apropos of the presumed place of transmission in Haiti for a child with conjunctival bancroftosis]. PMID- 10690479 TI - Statement concerning euthanasia and physician-assisted suicide. Ethics Committee of the College of Family Physicians of Canada. PMID- 10690480 TI - Viagra and broken hearts. PMID- 10690481 TI - Where are the Canadian data? PMID- 10690482 TI - Bosnia and Herzegovina. The challenge of change. PMID- 10690483 TI - Role and image of family physicians. Then and now. PMID- 10690484 TI - Managing women with nausea and vomiting of pregnancy. Canadian consensus. AB - QUESTION: I have a 30-year-old pregnant patient who is asthmatic and is taking prednisone. Whenever she decreases her prednisone dose, her nausea increases tremendously even though she is taking two tablets of doxylamine and pyridoxine combination (Diclectin) daily. She is already at 26 weeks' gestation, and I really do not want her to continue the prednisone beyond what is needed for her asthma, but two attempts to taper prednisone off failed because of unbearable nausea. ANSWER: Several controlled trials show the efficacy of prednisone for nausea and vomiting of pregnancy (NVP), but your case is fascinating in proving the point by challenge-rechallenge. You should try to decrease the prednisone while increasing doxylamine and pyridoxine to its recommended dose of two tablets before sleep, one in the morning, and on in the afternoon. PMID- 10690485 TI - Ophthaproblem. Acute angle-closure glaucoma. PMID- 10690486 TI - Palpitations. Deadly hens teeth. PMID- 10690487 TI - Practice tips. Endometrial biopsy. PMID- 10690488 TI - Walking or vigorous exercise? Which best helps prevent coronary heart disease in women? PMID- 10690489 TI - Olanzapine. Keep an eye on this neuroleptic. AB - Olanzapine (Zyprexa), a neuroleptic, has obtained marketing authorization for treatment of schizophrenia. The clinical file is satisfactory, but in the absence of relevant trials, it has not yet been demonstrated that olanzapine has a specific activity on the positive or negative symptoms of schizophrenia. The global efficacy of olanzapine was not substantially different from that of haloperidol in two of the three comparative trials published to date. The only relevant comparative trial fails to demonstrate the superiority of olanzapine over risperidone. Olanzapine has fewer adverse neurologic effects than haloperidol, but there is no evidence that it differs from other recent neuroleptics in this respect. Olanzapine can have anticholinergic adverse effects and frequently causes weight gain. Active surveillance is required because subclinical cases of elevated transaminase levels, increased blood pressure, and QT prolongation were observed in clinical trials (2500 patients treated). PMID- 10690490 TI - CPR or DNR? End-of-life decision making on a family practice teaching ward. AB - OBJECTIVE: To determine the proportion of patients on a family practice ward who had "code status" orders and end-of-life discussions documented on their charts in the first week of admission. To examine the correlation between a tool predicting the likelihood of benefit from cardiopulmonary resuscitation (CPR) and actual end-of-life decisions made by family physicians and their patients. DESIGN: Cross-sectional descriptive study using a retrospective chart review. SETTING: A 14-bed teaching ward where family physicians admit and manage their own patients in an urban tertiary care teaching hospital. PARTICIPANTS: Patients admitted to the ward for 7 or more days between December 1, 1995, and August 31, 1996. MAIN OUTCOME MEASURES: Frequency of documented "do not resuscitate" (DNR) or "full code" orders and documented end-of-life discussions. Prognosis-after resuscitation (PAR) score. RESULTS: In the 103 charts reviewed, code status orders were entered within 7 days for 60 patients (58%); 31 were DNR, and 29 were full code. Discussion of code status was documented in 25% of charts. The PAR score for 40% of patients was higher than 5, indicating they were unlikely to survive to discharge from hospital should they require CPR. There was a significant association between PAR scores done retrospectively and actual code status decisions made by attending family physicians (P < .005). CONCLUSIONS: End of-life discussions and decisions were not fully documented in patients' charts, even though patients were being cared for in hospital by their family physicians. A PAR score obtained during the first week of admission could assist physicians in discussing end-of-life orders with their patients. PMID- 10690491 TI - Preventive care for the elderly. Do family physicians comply with recommendations of the Canadian Task Force on Preventive Health Care? AB - OBJECTIVE: To assess to what extent family physicians perform the maneuvers for elderly patients recommended by the Canadian Task Force on Preventive Health Care (CTF), and to compare physicians' performance among patients who had structured periodic health examinations with performance among those who did not. DESIGN: Retrospective chart audit. SETTING: Family practice unit in a secondary care, university-affiliated hospital in Toronto, Ont. PARTICIPANTS: Records of 136 community-dwelling patients aged 70 and older. Of 340 randomly selected charts, 108 were excluded and 51 were inaccessible; 100 had had PHEs, and a random sample of 36 who had attended the clinic three or more times was chosen from the remaining 81 [corrected]. MAIN OUTCOME MEASURES: Proportion of patients who received the recommended screening maneuvers. RESULTS: Charts were audited for 100 patients who had structured periodic health examinations and 36 who did not but who attended the clinic three or more times during an 18-month period. Screening rates among patients who had structured examinations ranged from 28% of patients screened for hearing impairment to 100% screened for hypertension. Patients who did not have structured examinations were significantly less likely to receive screening maneuvers. CONCLUSIONS: Screening rates were below desirable levels in patients older than 70 years. Screening during structured health examinations seems to be more effective than opportunistic screening for patients 70 and older. PMID- 10690492 TI - Sons as sole caregivers for their elderly parents. How do they cope? AB - OBJECTIVE: To examine the experiences of men who are sole caregivers for their elderly parents. DESIGN: Semistructured in-depth interviews. SETTING: Family practice clinic attached to a large tertiary care centre in north central Toronto. PARTICIPANTS: A convenience sample of 10 men who identified themselves as sole caregivers in that they had no particular women assisting them with caregiving. METHOD: Interviews were analyzed by standard qualitative methods. MAIN FINDINGS: Emerging themes were the spectrum of caregiving, the experience of caregiving, and the use of formal support systems. Scope of care varied from very little to total care, including personal care. Participants described positive and negative aspects of and the nature of their relationships with those for whom they cared. Avoiding institutionalization was seen as positive; effects on work and social life were negative. Use of more than homemaking services was associated with previous hospitalization; participants complained about difficulties accessing services. CONCLUSIONS: The nature of sons' relationships with their parents and the amount of time they have available can predict how much caregiving they can undertake. Information about community support services is not readily accessible to these men. PMID- 10690493 TI - Surgical treatments for Parkinson's disease. AB - OBJECTIVE: This article reviews surgical treatments for Parkinson's disease, emphasizing aspects pertinent to family physicians: rationale for and description of surgeries, patient selection issues, and outcome expectations. QUALITY OF EVIDENCE: No published series describes long-term follow up of a randomized controlled study of any surgery for Parkinson's disease. Some reports, however, describe thorough but brief follow up of functioning in small numbers of patients following surgery. MEDLINE articles were identified using Parkinson's disease, surgery, pallidotomy, thalamotomy, stimulation, grafting, and transplantation as search words. Articles chosen for this paper described patients with systematic follow up using accepted validated rating scales. MAIN MESSAGE: Reported series show impressive improvements to patients undergoing lesioning, stimulation, and grafting surgery for Parkinson's disease. These patients are typically severely disabled but highly selected, and follow up is brief. Stereotactic lesioning (pallidotomy and thalamotomy), deep brain stimulation (thalamic, and elsewhere) and grafting (striatal) can be performed safely, but results vary greatly among centres. CONCLUSIONS: Certain Parkinson's disease patients might benefit from surgery. Ideal candidates for pallidotomy experience motor fluctuations with disabling levodopa-induced dyskinesias. Tremors resistant to antiparkinsonian medications sometimes respond to thalamotomy or thalamic stimulation. Other parkinsonian syndromes, dementias, and difficulties with gait and balance respond poorly to unilateral pallidotomy. Bilateral deep brain stimulation procedures could benefit "midline" dysfunction. PMID- 10690494 TI - Caregivers for people with dementia. What is the family physician's role? AB - OBJECTIVE: To examine the role of family physicians in providing support and care to caregivers for people with dementia. QUALITY OF EVIDENCE: Data were obtained from Alzheimer Society guidelines, published consensus statements, and guidelines for family physicians caring for people with dementia and their caregivers. Most of the reported findings and recommendations are based on information from expert consensus statements and opinion. MAIN MESSAGE: Caring for people with dementia causes substantial psychological and physical morbidity. Services developed for caregivers (in-home respite and individual psychological interventions) and comprehensive support programs are helpful in relieving caregiver distress. There is a role for family physicians in following caregivers longitudinally to assess their physical and emotional health and coping skills, to provide information and assistance in dealing with problems as they arise, to assist caregivers in mobilizing family and friends, and to facilitate referrals to appropriate services and resources. CONCLUSIONS: Family physicians have an important role in identifying caregiver problems and providing direct and ongoing support to caregivers in their day-to-day role. PMID- 10690495 TI - Managing benign prostatic hyperplasia in primary care. Patient-centred approach. AB - PROBLEM ADDRESSED: Management of benign prostatic hyperplasia (BPH) is changing from a surgical approach to a medical approach, and the role of primary care physicians is expanding. OBJECTIVE OF PROGRAM: To introduce a patient-centred approach to managing BPH in primary care through a continuing medical education (CME) program. MAIN COMPONENTS OF PROGRAM: A practice-based, small group, peer led CME program focused on application of the International Prostate Symptom Score and Quality of Life Assessment in four case studies on prostatism, including BPH. At 86 workshops held across Canada, 658 physicians participated in discussions with case materials that included videos and a handbook. A before after practice behaviour questionnaire was administered at each workshop to evaluate "intent to change." CONCLUSIONS: Participating physicians showed willingness to learn new skills for patient-centred management of BPH. These results suggest that peer-led, small group CME can successfully encourage use of new practice guidelines in primary are and teach physicians practical steps for developing therapeutic alliances with their patients. PMID- 10690496 TI - Managing sexual dysfunction. Using sildenafil for patients with cardiovascular disease. Heart and Stroke Foundation of Canada. Canadian Cardiovascular Society. PMID- 10690497 TI - Making choices. PMID- 10690498 TI - Hugh Davson--his contribution to the physiology of the cerebrospinal fluid and blood-brain barrier. PMID- 10690499 TI - Neural induction of the blood-brain barrier: still an enigma. AB - 1. The study of the blood-brain barrier and its various realms offers a myriad of opportunities for scientific exploration. This review focuses on two of these areas in particular: the induction of the blood-brain barrier and the molecular mechanisms underlying this developmental process. 2. The creation of the blood brain barrier is considered a specific step in the differentiation of cerebral capillary endothelial cells, resulting in a number of biochemical and functional alterations. Although the specific endothelial properties which maintain the homeostasis in the central nervous system necessary for neuronal function have been well described, the inductive mechanisms which trigger blood-brain barrier establishment in capillary endothelial cells are unknown. 3. The timetable of blood-brain barrier formation is still a matter of debate, caused largely by the use of varying experimental systems and by the general difficulty of quantitatively measuring the degree of blood-brain barrier "tightness." However, there is a general consensus that a gradual formation of the blood-brain barrier starts shortly after intraneural neovascularization and that the neural microenvironment (neurons and/or astrocytes) plays a key role in inducing blood brain barrier function in capillary endothelial cells. This view stems from numerous in vitro experiments using mostly cocultures of capillary endothelial cells and astrocytes and assays for easily measurable blood-brain barrier markers. In vivo, there are great difficulties in proving the inductive influence of the neuronal environment. Also dealt with in this article are brain tumors, the least understood in vivo systems, and the induction or noninduction of barrier function in the newly established tumor vascularization. 4. Finally, this review tries to elucidate the question concerning the nature of the inductive signal eliciting blood-brain barrier formation in the cerebral microvasculature. PMID- 10690500 TI - Barriers in the immature brain. AB - 1. The term "blood-brain barrier" describes a range of mechanisms that control the exchange of molecules between the internal environment of the brain and the rest of the body. 2. The underlying morphological feature of these barriers is the presence of tight junctions which are present between cerebral endothelial cells and between choroid plexus epithelial cells. These junctions are present in blood vessels in fetal brain and are effective in restricting entry of proteins from blood into brain and cerebrospinal fluid. However, some features of the junctions appear to mature during brain development. 3. Although proteins do not penetrate into the extracellular space of the immature brain, they do penetrate into cerebrospinal fluid by a mechanism that is considered in the accompanying review (Dziegielewska et al., 2000). 4. In the immature brain there are additional morphological barriers at the interface between cerebrospinal fluid and brain tissue: strap junctions at the inner neuroependymal surface and these and other intercellular membrane specializations at the outer (piaarachnoid) surface. These barriers disappear later in development and are absent in the adult. 5. There is a decline in permeability to low molecular weight lipid insoluble compounds during brain development which appears to be due mainly to a decrease in the intrinsic permeability of the blood-brain and blood-cerebrospinal fluid interfaces. PMID- 10690501 TI - The nature and composition of the internal environment of the developing brain. AB - 1. The fetal brain develops within its own environment, which is protected from free exchange of most molecules among its extracellular fluid, blood plasma, and cerebrospinal fluid (CSF) by a set of mechanisms described collectively as "brain barriers." 2. There are high concentrations of proteins in fetal CSF, which are due not to immaturity of the blood-CSF barrier (tight junctions between the epithelial cells of the choroid plexus), but to a specialized transcellular mechanism that specifically transfers some proteins across choroid plexus epithelial cells in the immature brain. 3. The proteins in CSF are excluded from the extracellular fluid of the immature brain by the presence of barriers at the CSF-brain interfaces on the inner and outer surfaces. These barriers are not present in the adult. 4. Some plasma proteins are present within the cells of the developing brain. Their presence may be explained by a combination of specific uptake from the CSF and synthesis in situ. 5. Information about the composition of the CSF (electrolytes as well as proteins) in the developing brain is of importance for the culture conditions used for experiments with fetal brain tissue in vitro, as neurons in the developing brain are exposed to relatively high concentrations of proteins only when they have cell surface membrane contact with CSF. 6. The developmental importance of high protein concentrations in CSF of the immature brain is not understood but may be involved in providing the physical force (colloid osmotic pressure) for expansion of the cerebral ventricles during brain development, as well as possibly having nutritive and specific cell development functions. PMID- 10690502 TI - Tight junctions of the blood-brain barrier. AB - 1. The blood-brain barrier is essential for the maintenance and regulation of the neural microenvironment. The blood-brain barrier endothelial cells comprise an extremely low rate of transcytotic vesicles and a restrictive paracellular diffusion barrier. The latter is realized by the tight junctions between the endothelial cells of the brain microvasculature, which are subject of this review. Morphologically, blood-brain barrier-tight junctions are more similar to epithelial tight junctions than to endothelial tight junctions in peripheral blood vessels. 2. Although blood-brain barrier-tight junctions share many characteristics with epithelial tight junctions, there are also essential differences. However, in contrast to tight junctions in epithelial systems, structural and functional characteristics of tight junctions in endothelial cells are highly sensitive to ambient factors. 3. Many ubiquitous molecular constituents of tight junctions have been identified and characterized including claudins, occludin, ZO-1, ZO-2, ZO-3, cingulin, and 7H6. Signaling pathways involved in tight junction regulation comprise, among others, G-proteins, serine, threonine, and tyrosine kinases, extra- and intracellular calcium levels, cAMP levels, proteases, and TNF alpha. Common to most of these pathways is the modulation of cytoskeletal elements which may define blood-brain barrier characteristics. Additionally, cross-talk between components of the tight junction- and the cadherin-catenin system suggests a close functional interdependence of the two cell-cell contact systems. 4. Recent studies were able to elucidate crucial aspects of the molecular basis of tight junction regulation. An integration of new results into previous morphological work is the central intention of this review. PMID- 10690503 TI - Transferrin and transferrin receptor function in brain barrier systems. AB - 1. Iron (Fe) is an essential component of virtually all types of cells and organisms. In plasma and interstitial fluids, Fe is carried by transferrin. Iron containing transferrin has a high affinity for the transferrin receptor, which is present on all cells with a requirement for Fe. The degree of expression of transferrin receptors on most types of cells is determined by the level of Fe supply and their rate of proliferation. 2. The brain, like other organs, requires Fe for metabolic processes and suffers from disturbed function when a Fe deficiency or excess occurs. Hence, the transport of Fe across brain barrier systems must be regulated. The interaction between transferrin and transferrin receptor appears to serve this function in the blood-brain, blood-CSF, and cellular-plasmalemma barriers. Transferrin is present in blood plasma and brain extracellular fluids, and the transferrin receptor is present on brain capillary endothelial cells, choroid plexus epithelial cells, neurons, and probably also glial cells. 3. The rate of Fe transport from plasma to brain is developmentally regulated, peaking in the first few weeks of postnatal life in the rat, after which it decreases rapidly to low values. Two mechanisms for Fe transport across the blood-brain barrier have been proposed. One is that the Fe-transferrin complex is transported intact across the capillary wall by receptor-mediated transcytosis. In the second, Fe transport is the result of receptor-mediated endocytosis of Fe-transferrin by capillary endothelial cells, followed by release of Fe from transferrin within the cell, recycling of transferrin to the blood, and transport of Fe into the brain. Current evidence indicates that although some transcytosis of transferrin does occur, the amount is quantitatively insufficient to account for the rate of Fe transport, and the majority of Fe transport probably occurs by the second of the above mechanisms. 4. An additional route of Fe and transferrin transport from the blood to the brain is via the blood-CSF barrier and from the CSF into the brain. Iron-containing transferrin is transported through the blood-CSF barrier by a mechanism that appears to be regulated by developmental stage and iron status. The transfer of transferrin from blood to CSF is higher than that of albumin, which may be due to the presence of transferrin receptors on choroid plexus epithelial cells so that transferrin can be transported across the cells by a receptor-mediated process as well as by nonselective mechanisms. 5. Transferrin receptors have been detected in neurons in vivo and in cultured glial cells. Transferrin is present in the brain interstitial fluid, and it is generally assumed that Fe which transverses the blood-brain barrier is rapidly bound by brain transferrin and can then be taken up by receptor-mediated endocytosis in brain cells. The uptake of transferrin-bound Fe by neurons and glial cells is probably regulated by the number of transferrin receptors present on cells, which changes during development and in conditions with an altered iron status. 6. This review focuses on the information available on the functions of transferrin and transferrin receptor with respect to Fe transport across the blood-brain and blood-CSF barriers and the cell membranes of neurons and glial cells. PMID- 10690505 TI - Can molecular assessment improve classification of head and neck premalignancy? PMID- 10690504 TI - The blood-brain barrier and bilirubin encephalopathy. AB - 1. The pathogenesis of bilirubin encephalopathy is multifactorial, involving the transport of bilirubin or albumin/bilirubin across the blood-brain barrier and delivering bilirubin to target neurons. 2. The relative importance of the blood brain barrier, unconjugated bilirubin levels, serum binding, and tissue susceptibility in this process is only partially understood. Even at dangerously high serum levels, bilirubin traverses the intact blood-brain barrier slowly, requiring time for encephalopathy to occur, although deposition of bilirubin can be rapid if a surge in plasma unbound bilirubin is produced by administering a drug which competes with bilirubin for binding to albumin. 3. There may be maturational changes in permeability both in the fetus and postnatally which protect the brain from bilirubin. 4. Disruption or partial disruption of the blood-brain barrier by disease or hypoxic ischemic injury will facilitate transport of bilirubin/albumin into brain, but the relative affinities of albumin and target neurons will determine whether the tissue bilirubin load is sufficient for toxicity to occur. PMID- 10690506 TI - Modulation of molecular targets to enhance radiation. PMID- 10690507 TI - Targeted toxins. AB - Targeted toxins, consisting of tumor-selective ligands coupled to polypeptide toxins, represent a new class of cancer therapeutics that kills malignant cells by inactivating cytosolic protein synthesis and inducing apoptosis. A number of these molecules have been produced under good manufacturing practice conditions and given systemically to patients with a variety of neoplasms. The promising results to date and the remaining pharmacological hurdles are discussed. PMID- 10690508 TI - Molecular determinants of response to TRAIL in killing of normal and cancer cells. AB - The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L) is a potent inducer of death of cancer but not normal cells, which suggests its potential use as a tumor-specific antineoplastic agent. TRAIL binds to the proapoptotic death receptors DR4 and the p53-regulated proapoptotic KILLER/DR5 as well as to the decoy receptors TRID and TRUNDD. In the present studies, we identified a subgroup of TRAIL-resistant cancer cell lines characterized by low or absent basal DR4 or high expression of the caspase activation inhibitor FLIP. Four of five TRAIL-sensitive cell lines expressed high levels of DR4 mRNA and protein, whereas six of six TRAIL-resistant cell lines expressed low or undetectable levels of DR4 (chi 2; P < 0.01). FLIP expression appeared elevated in five of six (83%) TRAIL-resistant cell lines and only one of five (20%) TRAIL sensitive cells (chi 2; P < 0.05). Two TRAIL-resistant lines that expressed DR4 contained an A-to-G alteration in the death domain encoding arginine instead of lysine at codon 441. The K441R polymorphism is present in 20% of the normal population and can inhibit DR4-mediated cell killing in a dominant-negative fashion. The expression level of KILLER/DR5, TRID, TRUNDD or TRID, and TRUNDD did not correlate with TRAIL sensitivity (P > 0.05). These results suggest that the major determinants for TRAIL sensitivity may be the expression level of DR4 and FLIP. TRAIL-resistant cells became susceptible to TRAIL-mediated apoptosis in the presence of doxorubicin. In TRAIL-sensitive cells, caspases 8, 9, and 3 were activated after TRAIL treatment, but in TRAIL-resistant cells, they were activated only by the combination of TRAIL and doxorubicin. Our results suggest: (a) evaluation of tumor DR4 and FLIP expression and host DR4 codon 441 status could be potentially useful predictors of TRAIL sensitivity, and (b) doxorubicin, in combination with TRAIL, may effectively promote caspase activation in TRAIL resistant tumors. PMID- 10690509 TI - Genetic progression and clonal relationship of recurrent premalignant head and neck lesions. AB - We constructed a preliminary genetic progression model for head and neck squamous cell carcinoma (HNSC) based on the frequency of genetic alterations in preneoplastic and neoplastic lesions from single biopsy specimens. To firmly establish the temporal order of established genetic events in HNSC, we sampled serial biopsies from five patients with recurrent premalignant lesions at a single anatomic site over a period of time (1 month to 144 months). These lesions were examined by microsatellite analysis of the minimal regions of loss on the 10 most frequently lost chromosomal arms in HNSC. Each set of serial biopsies from all five patients demonstrated LOH (loss of heterozygosity) of identical alleles at multiple loci with identical boundaries between areas of LOH and retention of heterozygosity, indicating a common clonal origin for each set. Three patients demonstrated genetic progression (new regions of LOH) over time correlating with histopathological progression, one patient demonstrated lack of genetic progression associated with unchanged histopathological morphology, and one patient demonstrated histopathological progression without detection of a corresponding genetic progression event. For one of these patients with a laryngeal tumor, at least four separate steps in progression to malignancy could be determined, accompanied by spatial expansion of an increasingly altered clonal population from the ipsilateral to the contralateral side, ultimately resulting in a malignancy. Microsatellite-based genetic analysis of recurrent premalignant lesions indicates that these lesions arise from a common clonal progenitor, followed by outgrowth of clonal populations associated with progressive genetic alterations and phenotypic progression to malignancy. PMID- 10690510 TI - Characterization of intracellular prostate-specific antigen from laser capture microdissected benign and malignant prostatic epithelium. AB - The proportion of unbound serum prostate-specific antigen (PSA; percent-free PSA) is reported to be lower in men with prostate cancer compared to men with benign prostates (U. H. Stenman et al., Cancer Res., 51: 222-226, 1991; H. Lilja et al., Clin. Chem., 37: 1618-1625, 1991; D. L. Woodrum et al., J. Urol., 159: 5-12, 1998; W. J. Catalona et al., J. Am. Med. Assoc., 279: 1542-1547, 1998). The majority of immunoreactive PSA in serum is complexed to alpha-1-antichymotrypsin (ACT). Two major mechanistic questions have previously been unknown: (a) Does PSA in human prostate cancer cells in tissue exist in a free or bound form? and (b) Is PSA produced by malignant cells in the free form because it has lost the ability to form a complex with ACT? Laser capture microdissection (LCM) enables the acquisition of pure populations of defined cell types from tissue (M. R. Emmert-Buck et al., Science, 274: 998-1001, 1996; R. F. Bonner et al., Science, 278: 1481-1483, 1997). This technology provides a unique opportunity to study intracellular protein composition and structure from human cells. In this study, we used LCM to assess the bound versus free form of intracellular PSA in both benign and malignant epithelium procured from prostate tissue. One-dimensional and two-dimensional PAGE were performed on cellular lysates from LCM-procured benign and malignant prostate epithelium from frozen tissue specimens. Western blotting analysis of one-dimensional PAGE gels revealed a strong band at M(r) 30,000 (expected molecular weight of unbound PSA) in all cases demonstrating that the vast majority of intracellular tumor and normal PSA exists within cells in the "free" form. Binding studies showed that PSA recovered from LCM-procured cells retained the full ability to bind ACT, and two-dimensional PAGE Western analysis demonstrated that the PSA/ACT complex was stable under strong reducing conditions. We conclude that intracellular PSA exists in the "free" form and that binding to ACT occurs exclusively outside of the cell. PMID- 10690511 TI - Use of allelic loss to predict malignant risk for low-grade oral epithelial dysplasia. AB - One of the best approaches to identifying genetic changes critical to oral cancer progression is to compare progressing and nonprogressing oral premalignant lesions. However, such samples are rare, and they require long-term follow-up. The current study used the large archive network and clinical database in British Columbia to study loss of heterozygosity (LOH) in cases of early oral premalignancies, comparing those with a history of progression to carcinoma in situ or invasive cancer and those without a history of progression (referred to as nonprogressing cases). Each of 116 cases was analyzed for LOH at 19 microsatellite loci on seven chromosome arms (3p, 4q, 8p, 9p, 11q, 13q, and 17p). The progressing and nonprogressing cases showed dramatically different LOH patterns of multiple allelic losses. An essential step for progression seems to involve LOH at 3p and/or 9p because virtually all progressing cases showed such loss. However, LOH at 3p and/or 9p also occurred in nonprogressing cases. Individuals with LOH at 3p and/or 9p but at no other arms exhibit only a slight increase of 3.8-fold in relative risk for developing cancer. In contrast, individuals with additional losses (on 4q, 8p, 11q, or 17p), which appeared uncommon in nonprogressing cases, showed 33-fold increases in relative cancer risk. In conclusion, analysis of LOH at 3p and 9p could serve as an initial screening for cancer risk of early premalignancies. Follow-up investigation for additional losses would be essential for predicting cancer progression. PMID- 10690512 TI - Successful surgical removal of occult metastases of medullary thyroid carcinoma recurrences with the help of immunoscintigraphy and radioimmunoguided surgery. AB - Patients with recurrent or metastatic medullary thyroid carcinoma (MTC) were referred for pretargeted immunoscintigraphy (Affinity Enhancement System; AES) and radioimmunoguided surgery (RIGS). Data collected from 13 patients establish that whole-body AES immunoscintigraphy revealed metastases < 360 mg and RIGS detected micrometastases (5-15 mg). All tissue samples removed by the surgeon were diagnosed by histology and immunohistochemistry of calcitonin to check the accuracy of IS and RIGS results. AES immunoscintigraphy is very sensitive. Of 34 metastases or recurrences detected, 22 had escaped physical examination or conventional imaging. The accuracy of RIGS was 86%, its sensitivity 75%, and its specificity was 90% (n = 208). IS and RIGS detected occult tumors that would have escaped surgery, clearly demonstrating clinical benefit. Serum calcitonin (normal, 10 pg/ml) and carcinoembryonic antigen (normal, 5 ng/ml) of two patients were restored to normal. In patients whose tumors were discovered, progression of their disease was slowed, as evidenced by the large decrease in serum calcitonin and carcinoembryonic antigen, an important prognostic factor. Surgery was canceled in one case where IS detected distant metastases out of surgical reach. Thus, AES immunoscintigraphy and RIGS might be of valuable help for the surgical management of medullary thyroid carcinoma. PMID- 10690513 TI - Molecular remission induction with retinoic acid and anti-CD33 monoclonal antibody HuM195 in acute promyelocytic leukemia. AB - Despite achieving complete remission with retinoic acid (RA), most patients with acute promyelocytic leukemia (APL) have minimal residual disease detectable by reverse transcription-PCR (RT-PCR) amplification. HuM195, a humanized monoclonal antibody reactive with the cell surface antigen CD33, specifically targets and kills myeloid leukemia cells. We studied whether HuM195 could eliminate minimal residual disease in patients with APL by using RT-PCR. After attaining clinical complete remission with RA and/or chemotherapy, patients received HuM195 twice weekly for 3 weeks. Patients in first remission were given consolidation chemotherapy, generally with three cycles of idarubicin and cytarabine. Patients in second or greater remission did not receive chemotherapy. All patients received six monthly courses of maintenance with two doses of HuM195. Twenty-five of 27 patients treated in first remission had positive RT-PCR determinations before HuM195 treatment. Of the 22 patients evaluable for conversion of positive RT-PCR assays, 11 (50%) became RT-PCR negative after HuM195 treatment without additional therapy. Within the subset of patients who received RA alone as induction, 8 of 18 evaluable patients (44%) had negative RT-PCR determinations after HuM195 treatment but before chemotherapy. Among similar patients treated on earlier studies, 7 of 34 patients (21%) induced into remission with RA and then maintained on the drug for 1 month were RT-PCR negative before chemotherapy (P = 0.07). Twenty-five of 27 patients with newly diagnosed APL (93%) remain in clinical complete remission for 7+ to 58+ months, with median follow-up of 29 months. Seven patients in second or third remission and one patient in molecular relapse were also treated. Only one of these patients became RT-PCR negative after treatment with HuM195. These data suggest that HuM195 has activity against minimal residual disease in APL, particularly in newly diagnosed patients. PMID- 10690514 TI - Angiogenesis, thymidine phosphorylase, and resistance of squamous cell head and neck cancer to cytotoxic and radiation therapy. AB - Thymidine phosphorylase (TP), an enzyme involved in the thymidine synthesis and degradation, has been shown to promote tumor angiogenesis. Both TP expression and tumor vascularization are putative postoperative prognostic markers of cancer. Because of its bifunctional role, TP may have interactions with cytotoxic drugs or radiation via pathways requiring thymidine or prodrug activation. The microvessel score and TP expression were examined immunohistochemically on paraffin-embedded bioptical material from 94 locally advanced squamous cell head and neck carcinomas. All patients were treated with conventionally fractionated radiotherapy combined with induction (platinum- and 5-fluorouracil-based) or concurrent platinum chemotherapy. The follow-up of patients ranged from 6 to 108 months (median, 48 months). Nuclear TP expression was significantly associated with increased microvessel score (P < 0.0001, r = 0.45). A low percentage of cancer cells with nuclear TP expression in pretreatment biopsies was associated with a high rate of CR after combined chemoradiotherapy (P = 0.006) and induction chemotherapy (0.01). A better local relapse-free and overall survival was also observed in these patients (P = 0.001 and P = 0.0005, respectively). Biospies on the day after the delivery of 20 Gy of conventionally fractionated radiotherapy showed residual cancer cell nests, frequently of high vascularization and of intense nuclear TP reactivity. It is concluded that thymidine phosphorylase is associated with angiogenesis, with resistance to radiotherapy and cytotoxic therapy, and with poorer survival in squamous cell head and neck cancer. A strong rationale is provided for subsequent clinical trials of concurrent radiotherapy and chemotherapy with antiangiogenic agents or with specific TP inhibitors. PMID- 10690515 TI - Phase I clinical and pharmacokinetic study of perillyl alcohol administered four times a day. AB - We conducted a phase I dose-escalation trial of perillyl alcohol (POH; NSC 641066) given p.o. on a continuous four times a day basis to characterize the maximum tolerated dose, toxicities, pharmacokinetic profile, and antitumor activity. Sixteen evaluable patients with advanced refractory malignancies were treated at the following doses: level 1 (L1), 800 mg/m2/dose; L2, 1200 mg/m2/dose; L3, 1600 mg/m2/dose. POH was formulated in soft gelatin capsules containing 250 mg of POH and 250 mg of soybean oil. The predominant toxicities seen were gastrointestinal (nausea, vomiting, satiety, and eructation), which were dose limiting. There appeared to be a dose-dependent increase in levels of the two main metabolites, perillic acid and dihydroperillic acid. No significant differences were seen whether the drug was taken with or without food. There was a trend toward decreasing metabolite levels on day 29 compared with days 1 and 2. Peak metabolite levels were seen 1-3 h post ingestion. Metabolite half-lives were approximately 2 h. Approximately 9% of the total dose was recovered in the urine in the first 24 h, the majority as perillic acid. Evidence of antitumor activity was seen in a patient with metastatic colorectal cancer who has an ongoing near complete response of > 2 years duration. Several other patients were on study for > or = 6 months with stable disease. The maximum tolerated dose of POH given continuously four times a day was 1200 mg/m2/dose. Gastrointestinal toxicity was dose limiting, although significant interpatient variability in drug tolerance was seen. PMID- 10690516 TI - Phase I study of ONO-4007, a synthetic analogue of the lipid A moiety of bacterial lipopolysaccharide. AB - ONO-4007 is a synthetic analogue of the lipid A moiety of bacterial lipopolysaccharide, which exhibits antitumor activity by the induction of intratumoral tumor necrosis factor alpha, the potentiation of tumor-infiltrating macrophages, and the inhibition of angiogenesis. Interleukin (IL)-1 alpha, IL-6, and IL-12 induction by ONO-4007 activates cytotoxic natural killer cells to up regulate IFN-gamma and nitric oxide synthase activity. ONO-4007 was given to 24 patients (13 males and 11 females; median age, 53 years) as a 30-min i.v. infusion on day 1, followed on day 15 by a first treatment cycle consisting of three weekly infusions at the same dose, followed by a rest period of 1 week. Cohorts of six patients received up to a maximum of four treatment cycles at increasing dose levels (75, 100, and 125 mg). The maximum tolerated dose was 125 mg, with grade 3 National Cancer Institute Common Toxicity Criteria toxicity (rigors with cyanosis) occurring in two of six patients at this dose level. An additional six patients were treated at 100 mg, the dose below the maximum tolerated dose. Other toxicities included grade 2 National Cancer Institute Common Toxicity Criteria myalgia, nausea, and hypotension. The pharmacokinetics of ONO-4007 appeared to be independent of dose and showed linearity with respect to time. ONO-4007 has a low systemic clearance (approximately 1.3 ml/min) and a small volume of distribution (5-8 liters) with a long t1/2 of 74-95 h. The administration of ONO-4007 was shown to result in a significant increase in circulating levels of tumor necrosis factor alpha and IL-6. No objective antitumor responses were observed. Seven patients maintained stable disease for at least two cycles, whereas five patients maintained stable disease for the full four-cycle duration of the study. Additional studies are required to determine the antitumor activity of ONO-4007. PMID- 10690517 TI - Phase II trial of yttrium-90-DOTA-biotin pretargeted by NR-LU-10 antibody/streptavidin in patients with metastatic colon cancer. AB - A Phase II study of yttrium-90-tetra-azacyclododecanetetra-acetic acid-biotin (90Y-DOTA-biotin) pretargeted by NR-LU-10 antibody/streptavidin (SA) was performed. The primary objectives of the study were to evaluate the efficacy and safety of this therapy in patients with metastatic colon cancer. Twenty-five patients were treated with a single dose of 110 mCi/m2 (mean administered dose, 106.5 +/- 10.3 mCi/m2) of 90Y-DOTA-biotin. There were three components of the therapy. Patients first received NR-LU-10/SA on day 1. A clearing agent (biotin galactose-human serum albumin) was administered approximately 48 h after the NR LU-10/SA to remove residual circulating unbound NR-LU-10/SA. Lastly, 24 h after administration of clearing agent, patients received biotin-DOTA-labeled with 110 mCi/m2 90Y. All three components of the therapy were administered i.v. Both hematological and nonhematological toxicities were observed. Diarrhea was the most frequent grade 4 nonhematological toxicity (16%; with 16% grade 3 diarrhea). Hematological toxicity was less severe with 8% grade 3 and 8% grade 4 neutropenia and 8% grade 3 and 16% grade 4 thrombocytopenia. The overall response rate was 8%. Two partial responders had freedom from progression of 16 weeks. Four patients (16%) had stable disease with freedom from progression of 10-20 weeks. Despite the relatively disappointing results of this study in terms of therapeutic efficacy and toxicity, proof of principle was obtained for the pretargeting approach. In addition, valuable new information was obtained about normal tissue tolerance to low-dose-rate irradiation that will help to provide useful guidelines for future study designs. PMID- 10690518 TI - Modulation of clinical drug resistance in a B cell lymphoma patient by the protein kinase inhibitor 7-hydroxystaurosporine: presentation of a novel therapeutic paradigm. AB - Emerging evidence suggests that apoptosis is an important mechanism of tumor cell death from antineoplastic therapy. 7-hydroxystaurosporine (UCN-01) is a novel protein kinase inhibitor that increases chemotherapy-induced apoptosis in vitro and is in early phases of clinical development. In this report, we present a 68 year-old patient with chemotherapy-resistant lymphoma treated with UCN-01 and chemotherapy. He had a stage IV plasmacytoid lymphoma that failed to enter remission with high-dose EPOCH II (etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin) chemotherapy. Due to disease progression and transformation to large cell lymphoma in the liver and bone marrow, he received UCN-01. Four weeks later, he received "standard-dose" EPOCH because of progression, developed severe neutropenia for 9 days, and expired from Candida sepsis on day 23. At autopsy, there was no histological evidence of residual lymphoma, although PCR for immunoglobulin gene rearrangement analysis revealed a faint clonal band in two of six nodes but none in the liver. Significantly, no B cells were detected by immunohistochemistry in lymph nodes, and a polyclonal ladder pattern associated with the presence of normal B cells was not seen in the immunoglobulin gene rearrangement PCR assay. Profound peripheral lymphopenia (50 cells/microliter) was also observed. Pharmacokinetics showed UCN-01 salivary concentrations, a surrogate for free drug concentrations, to be within an effective range in vitro (45 nmol/L) as a modulator of DNA-damaging agent cytotoxicity. In vitro, UCN-01 is synergistic with multiple cytotoxic agents and increases fludarabine-induced apoptosis in a human breast cell line. These results suggest that UCN-01 sensitized the lymphoma to the cytotoxic effects of EPOCH, possibly by modulating the "threshold" for apoptosis, and may illustrate a new paradigm for reversal of drug resistance. PMID- 10690519 TI - A neoadjuvant clinical trial in colorectal cancer patients of the human anti idiotypic antibody 105AD7, which mimics CD55. AB - Thirty-five patients received 105AD7 human anti-idiotype vaccination prior to surgery for colorectal carcinoma. Patients were immunized before and also received one to two immunizations after surgical resection of their colorectal cancer. The vaccine was well tolerated with no associated toxicity. Lymphocytic infiltration within the resected tumors was quantified by immunohistochemistry and image analysis. Enhanced infiltration of helper T cells (CD4) and natural killer (NK) cells (CD56) were observed in the tumors from immunized patients when compared with tumors from stage, grade, site, age, and sex matched unimmunized patients. NK activity was increased in the blood, peaking 7-10 days post immunization and then dropping rapidly and correlating with NK extravasation within the tumor. Comparison of the amino acid sequences of 105AD7 anti-idiotype and the antigen it mimics, CD55, has predicted that patients with HLA-DR1, HLA DR3, and HLA-DR7 haplotypes should show helper T cell responses following 105AD7 vaccination. Eighty-three percent of patients expressing these haplotypes responded to 105AD7, whereas 88% of patients who failed to express these haplotypes were nonresponders. With a median follow-up of 4 years (range, 2.5-6 years) 65% of patients remained disease free. This trial shows that 105AD7 stimulates antitumor inflammatory responses allowing extravasation within tumor deposits of both helper T cells and NK cells. This represents a way of evaluating immune responses in patients both within the blood and at the tumor site. The study confirms that immunization with a human anti-idiotypic antibody results in immune responses in 83% of patients with a permissive haplotype. PMID- 10690520 TI - Effect of food on the pharmacokinetics of oral MMI270B (CGS 27023A), a novel matrix metalloproteinase inhibitor. AB - MMI270B is a matrix metalloproteinase inhibitor (MMPI) with in vitro and in vivo activity. To exert optimal target inhibition, MMPI must be given chronically, and therefore, oral bioavailability is important. We analyzed the effect of food intake on AUC0-8 h, Cmax, and Tmax. Seventeen patients were entered into the study. Doses of MMI270B were 150, 400, and 600 mg. The first day, patients ingested the drug in a fasted state and were not allowed to eat for 2 h. The second day, patients ingested the drug 30 min after a light breakfast. Mean AUC0 8 h was not significantly influenced by food intake. Plasma concentrations were well above the IC50 of several MMPs at all doses tested. Mean Cmax was significantly decreased after food intake. Mean Tmax was significantly delayed after food intake. Food intake did not result in a significant change in exposure to MMI270B (AUC0-8 h) but did result in a significant, although not clinically relevant, decrease in peak plasma levels and time to reach peak plasma levels. No specific guidelines concerning the ingestion of MMI270B in either a fed or a fasted state are recommended. PMID- 10690521 TI - Detection and analysis of cancer cells in blood and bone marrow using a rare event imaging system. AB - An automated rare event detection system (Rare Event Imaging System) is described for the recognition of cancer cells that appear at low frequencies (1 in 1 million) in peripheral blood (PB) or bone marrow (BM). The instrumentation includes an automated fluorescence microscope (Nikon Microphot-FXA) with a cooled charge coupled device camera and a 60-MHz Pentium personal computer. Main features of the system are rapid analysis of large microscopic fields, including a total cell count, detection of fluorescently labeled cells, and a display of digitally stored images of the detected cells. Furthermore, the X,Y coordinates of each identified object are stored and can be recalled for morphological analysis of the cell using higher magnification or different fluorescent filter sets. The preparation of the blood or BM samples for automated analysis consists of lysis of the RBCs, attachment of sample cells onto adhesion slides, fixation, and fluorescent labeling with anticytokeratin antibodies. Cytokeratin-positive cells, however, were detected in 17% of the samples from healthy blood donors using this procedure (mean number, approximately 7/10(6) mononuclear cells in positive samples). To improve the specificity of the rare event detection, a double-labeling protocol combining intracellular cytokeratin with epithelial cell adhesion molecule (Ep-CAM) (breast, ovarian, colon, and lung carcinoma antigen) or disialo-ganglioside (GD2) antigen (small cell lung carcinoma, neuroblastoma, melanoma antigen) was developed. Examples of doubly labeled cultured cells and cancer cells from breast and small cell lung cancer patients are shown. Using the double-labeling protocol, no "positive" cells were seen in samples of healthy blood donors. Automated rare event detection (cytokeratin single-staining) was applied to 355 PB, BM, and stem cell (SC) samples from breast cancer patients before autologous BM transplantation. Cytokeratin-positive cells were found in 52% of BM, 35% of PB, and 27% of SC samples at frequencies of 1-1020 positive cells/10(6) mononuclear cells, thereby establishing the efficacy of the technique in the detection of rare cancer cells in hematopoietic tissue samples of cancer patients. PMID- 10690522 TI - Prognostic significance of mutations to different structural and functional regions of the p53 gene in breast cancer. AB - Alteration to the p53 tumor suppressor gene is associated with more aggressive disease in breast cancer, as evidenced by the shortened survival of patients with mutation. Data obtained from in vitro experiments suggest that mutations to different structural and functional domains of p53 may give rise to different effects on its biological activities, notably transactivational and apoptotic properties. We evaluated the prognostic significance of various types of p53 mutation in a series of 178 tumors identified by PCR-single-strand conformational polymorphism screening as containing a mutant gene. Mutations within exon 4 were associated with particularly poor prognosis, possibly relating to the importance of this region in apoptosis. Mutations that caused denaturation of the protein structure were also associated with poor survival, again perhaps because of effects on apoptosis. In contrast, patients with mutations in the DNA contact region showed similar survival to that of patients with normal p53, suggesting a less important role for p53-mediated transactivation in determining tumor aggressiveness. Other mutation groups associated with poor prognosis were single base substitutions and transversion mutations. Mutations in exon 6, exon 7, or the "hotspot" codons (175, 245, 248, 273) were associated with only a small reduction in patient survival compared with normal p53. These results allow some insight to be gained into the functional importance of various p53 domains in terms of their influence on overall patient survival. Further work is required to determine whether these domains are also important in influencing the response of breast tumors to adjuvant therapies. PMID- 10690523 TI - Quantitation of hTERT gene expression in sporadic breast tumors with a real-time reverse transcription-polymerase chain reaction assay. AB - Recent observations support the notion that telomerase expression is essential for the formation of human tumor cells [W-C. Hahn et al., Nature (Lond.), 400: 464-468, 1999]. The expression pattern of hTERT, the human telomerase catalytic subunit gene, is a rate-limiting determinant of the enzymatic activity of human telomerase. We have developed a real-time quantitative RT-PCR assay based on Taq Man fluorescence methodology to quantify the full range of hTERT mRNA copy numbers. We validated the method on a series of 134 unilateral invasive primary breast cancer patients with known long-term outcome. Three-quarters of the breast tumors (75.4%; 101 of 134) were hTERT positive, i.e., contained detectable and quantifiable hTERT mRNA. hTERT-positive patients had significantly shorter relapse-free survival (P = 0.017) after surgery compared with hTERT-negative patients. The prognostic significance of hTERT status persisted in Cox multivariate regression analysis. When we subdivided hTERT-positive patients (n = 101) into three equal groups (tumors showing small, intermediate, or high increase in hTERT mRNA content), we observed statistical (or a trend toward) links between high hTERT mRNA levels and Scarff-Bloom-Richardson histopathological grade III (P = 0.066), and negative estrogen (P = 0.002) and progesterone (P = 0.048) receptor status, and therefore with higher aggressiveness of breast tumors. High hTERT mRNA levels were also linked to MYC gene overexpression (P = 0.007). These findings show that the quantitative evaluation of hTERT mRNA can have important prognostic significance in human breast cancer. In addition, our simple, rapid, and semiautomated assay method is suitable for routine hTERT mRNA detection and quantification and will be a powerful tool in large, randomized, prospective, cooperative group trials and in the hTERT-based therapy project. PMID- 10690524 TI - Altered expression of beta-catenin in renal cell cancer and transitional cell cancer with the absence of beta-catenin gene mutations. AB - Loss of normal beta-catenin expression and the beta-catenin gene mutations have been shown to contribute to the malignant character of various cancers. Using PCR single-strand conformation polymorphism and DNA direct sequencing, we examined the presence of genetic alterations within the third exon of beta-catenin, which are frequently observed in other tumors, in transitional cell cancer (TCC) and renal cell cancer (RCC) cell lines, and in tumor specimens. The degrees of expression and intracellular distribution of beta-catenin were detected by immunohistochemical staining in 77 primary and 12 metastatic RCCs and in 81 primary TCCs. Western blot analysis was also applied to confirm the degree of beta-catenin expression in the cell lines and some tumor samples. We failed to reveal any genetic alterations, at least in the third exon of the beta-catenin gene, in RCC and TCC. Reduced membranous immunoreactivity of beta-catenin was observed in portions of RCC (15.5%) and TCC (24.7%) and was correlated with advanced stages and nodal involvement in RCC and with advanced stages and multiple tumors in TCC. Within the power limitations of this small study, beta catenin abnormal expression was not correlated with recurrence or survival in either RCC or TCC. Interstitial deletions and mutations in the third exon of beta catenin do not play a significant role in RCC or TCC tumorigenesis. Down regulation of normal beta-catenin expression might contribute to the malignant character of RCC and TCC and result in tumor progression. However, this event is not an independent prognostic factor for recurrence or tumor specific survival. PMID- 10690525 TI - Serum total and free prostate-specific antigen for breast cancer diagnosis in women. AB - Prostate-specific antigen (PSA) is a serine protease expressed at high levels in prostate epithelium, and elevated PSA in serum is a well-established marker of prostate cancer. Recently, the relative proportions of free PSA and PSA complexed to the serine protease inhibitor alpha 1-antichymotrypsin have become important variables in distinguishing between prostate cancer and benign prostatic hyperplasia. Numerous studies have demonstrated the production of PSA in female tissues such as the breast, and low levels of PSA are present in female sera. The objective of this study was to measure and compare the relative proportions of free PSA and PSA complexed to the serine protease inhibitor alpha 1 antichymotrypsin in the serum of women with breast cancer or benign breast disease or women with no known malignancies. PSA was measured with an established immunoassay for total PSA and a novel immunoassay for free PSA, both of which had a detection limit of 0.001 microgram/liter (1 ng/liter). The percentage of breast cancer patients with free PSA as the predominant molecular form (> 50% of total PSA) in serum was five times higher than that of healthy women or women with benign breast disease, and PSA decreased in the serum of breast cancer patients after surgery. The diagnostic use of free PSA for breast cancer is limited at this point, due to the low diagnostic sensitivity (approximately 20%); however, free PSA as the predominant molecular form shows a high diagnostic specificity (approximately 96%) in comparison to women free of breast cancer or with benign breast disease. These results suggest that the clinical applicability of free PSA for breast cancer diagnosis and the biological mechanism behind its increase should be further investigated. PMID- 10690526 TI - Loss of imprinting of the IGF-II and H19 genes in epithelial ovarian cancer. AB - To establish a possible role of genomic imprinting in the carcinogenesis of epithelial ovarian cancer, we determined the imprinting status of both IGF-II and H19 genes in 43 ovarian cancers, 7 low malignant potential ovarian tumors, and their matched normal tissues. In ovarian cancer, loss of heterozygosity (LOH) of IGF-II, H19 RsaI, and H19 AluI was found in 4 of 24 (16.7%), 3 of 20 (15%), and 1 of 16 (6.3%) samples, respectively. All patients with tumor specimens exhibiting LOH are of advanced clinical stages. Loss of imprinting (LOI) was found in 5 of 20 (25%) for IGF-II and in 4 of 17 (23.5%) and 1 of 15 (6.7%) for the RsaI and AluI sites of H19 gene with no LOH. However, no LOH was found in low malignant potential tumors, and only one of them showed LOI in H19 AluI site. Overexpression of IGF-II was demonstrated in all five LOI samples with normal expression of H19. Three of the five tumor specimens exhibiting LOI were transcribed from P1 promoter, whereas the remaining two were from the P3 promoter. These results suggested that LOH of both IGF-II and H19 genes was associated with advanced ovarian cancer. LOI of IGF-II and H19 genes may be involved in the development of ovarian cancer. Transcription of IGF-II from the P1 promoter may account for the biallelic expression of the IGF-II gene. PMID- 10690527 TI - Epigenetic regulation of gene expression in cervical cancer cells by the tumor microenvironment. AB - Evidence is accumulating that the adverse tumor microenvironment both modifies the malignant progression of tumor cells and contributes to chemotherapy and radiation resistance. We hypothesized that some of the effects on malignant progression are mediated through the transcriptional regulation of genes responsive to the stresses of the microenvironment, such as low oxygen or low glucose conditions. To determine epigenetic changes in gene expression that were consistent with that hypothesis, we used an in vitro subtractive hybridization method, representational difference analysis, to identify hypoxia-induced cDNAs from cultured human cervical epithelial cells. We identified 12 induced genes: two novel genes (HIG1 and HIG2), three genes known to be hypoxia-inducible (tissue factor, GAPDH, thioredoxin), and seven genes not previously identified as hypoxia-inducible [HNRNP(a1), ribosomal L7, annexin V, lipocortin 2, Ku(70), PRPP synthase, and acetoacetyl-CoA thiolase]. In cultured cells, HIG1 and HIG2 expression is induced by hypoxia and by glucose deprivation, but their expression is not induced by serum deprivation, UV, or ionizing radiation. The putative HIG1 and HIG2 open reading frames are expressed in cells, as confirmed by epitope tagging. In addition, tumor xenografts derived from human cervical cancer cells display increased expression of HIG1 and HIG2 when they are deprived of oxygen. Taken together, these data suggest a coordinated transcriptional response of eukaryotic cells to microenvironmental stresses found in the solid tumor. PMID- 10690528 TI - Immunohistochemically detected thymidylate synthase in colorectal cancer: an independent prognostic factor of survival. AB - Intratumoral thymidylate synthase (TS) expression and M(r) 53,000 phosphoprotein (p53) overexpression were studied immunohistochemically in sections from stored paraffin-embedded primary colorectal cancers in 70 patients who had undergone surgery during the years 1987-1990. These cancers were classified according to Dukes' stage A-D, using monoclonal antibodies TS 106 and DO-7. In patients with Dukes' stage A-C tumors, univariate analyses showed that there was a significant correlation (P = 0.048) between disease-free survival and TS expression and between TS expression and time to death with colorectal cancer (P = 0.038). In patients with Dukes' stage A-D tumors, overall survival was correlated to TS expression (P = 0.015), Dukes' stage (P < 0.001), and level of tumor differentiation (P = 0.044) but not to p53 overexpression. Patients with low intratumoral TS expression survived significantly longer than patients with high expression. Cox multivariate analysis showed that Dukes' stage (P < 0.001) and TS expression (P = 0.043) could independently serve as prognostic factors for time to death with colorectal cancer in patients with Dukes' stage A-D tumors. PMID- 10690529 TI - Enhanced GBX2 expression stimulates growth of human prostate cancer cells via transcriptional up-regulation of the interleukin 6 gene. AB - Previous studies demonstrated that the GBX2 homeobox gene is consistently overexpressed in cultured human prostate cancer cell lines. In this study, the human GBX2 cDNA was cloned and a quantitative reverse transcription-PCR method used to demonstrate that GBX2 mRNA expression is enhanced in approximately 70% of human prostate cancer tissues compared with normal human prostate tissues. Purified recombinant GBX2 protein binds specifically to an ATTA motif within the promoter of the interleukin 6 (IL-6) gene. Using an antisense approach, down regulation of the expression of GBX2 correlated with decreased expression of IL-6 and an inhibition of tumorigenicity of PC3 human prostate cancer cells. In addition, in vitro growth of the antisense clones was partially restored by exogenous addition of recombinant IL-6 protein to the culture media. These data demonstrated that enhanced GBX2 expression results in a stimulation of malignant growth of prostate cancer cells and that part of this stimulation involves up regulation in the transcription of the IL-6 gene. PMID- 10690530 TI - Vitamin D receptor polymorphisms are associated with altered prognosis in patients with malignant melanoma. AB - Calcitriol [1,25(OH)2D3], the hormonal derivative of vitamin D3, is an antiproliferative and prodifferentiation factor for several cell types, including cultured melanocytes and malignant melanoma (MM) cells. Several polymorphisms of the vitamin D receptor (VDR) gene have been described including a FokI RFLP in exon 2, BsmI, and ApaI polymorphisms in intron 8 and an adjacent TaqI RFLP in exon 9. Alterations in vitamin D/1,25(OH)2D3 levels and polymorphisms of the VDR have been shown to be associated with several systemic malignancies. We hypothesize that polymorphism in this gene may be associated with altered susceptibility and outcome in patients with MM. A hospital-based case-control study, using 316 MM cases and 108 controls, was used to assess associations with MM susceptibility. Breslow thickness, the most important single prognostic factor in MM, was used as the outcome measure. Polymorphisms at the FokI and TaqI restriction sites were determined using PCR-based methods. Polymorphism at the FokI, but not TaqI, RFLP was associated with an altered risk of MM (P = 0.014). More importantly, variant alleles were associated with increased Breslow thickness. Thus, homozygosity for variant alleles at both RFLP (ttff genotype combination) was significantly associated with thicker tumors. (> or = 3.5 mm; P = 0.001; odds ratio = 31.5). Thus, polymorphisms of the VDR gene, which would be expected to result in impaired function, are associated with susceptibility and prognosis in MM. These data suggest that 1,25(OH)2D3, the ligand of the VDR, may have a protective influence in MM, as has been proposed for other malignancies. PMID- 10690531 TI - Cysteine proteinase inhibitors stefin A, stefin B, and cystatin C in sera from patients with colorectal cancer: relation to prognosis. AB - The levels of cysteine proteinase inhibitors stefin A, stefin B, and cystatin C were determined using ELISAs in sera obtained preoperatively from 345 patients with colorectal cancer and in control sera from 125 healthy blood donors. The levels of stefin A and cystatin C were found to be moderately increased in patient sera (1.4-fold and 1.6-fold, respectively; P < 0.0001), whereas the level of stefin B remained statistically unchanged when compared with controls. The medians were 4.3 ng/ml versus 3.2 ng/ml for stefin A, 1.2 ng/ml versus 1.7 ng/ml for stefin B, and 679 ng/ml versus 425 ng/ml for cystatin C. In patient sera, a weak correlation of cystatin C with age (r = 0.34; P < 0.001) and gender (P = 0.01) was found. Stefin A and cystatin C levels were independent of Dukes' stage, whereas stefin B correlated significantly with Dukes' stage, its level being the highest in stage D (P < 0.007). Stefin B and cystatin C correlated with survival, whereas stefin A was not a significant prognostic factor in this study. Using medians as cutoff values, patients with high levels of stefin B and patients with high levels of cystatin C exhibited a significantly higher risk of death than those with low levels of inhibitors (hazard ratio = 1.6; 95% confidence interval, 1.2-2.2; P = 0.002 for stefin B; hazard ratio = 1.3; 95% confidence interval, 1.0 1.8; P = 0.04 for cystatin C). Our results reveal a correlation between high levels of extracellular cysteine proteinase inhibitors and short survival in patients with colorectal cancer, and the data thus support previous studies suggesting a contributing role of protease inhibitors in the progression of cancer. PMID- 10690532 TI - Expression levels of estrogen receptor-alpha, estrogen receptor-beta, coactivators, and corepressors in breast cancer. AB - Recent studies have indicated that a complex machinery of transactivation of target genes by estrogen or antiestrogen through estrogen receptor (ER) exists. However, the substantial roles of ER-beta, coactivators, and corepressors in the development and progression of breast cancer remain to be elucidated. To obtain some clue to these roles, we screened the expression levels of ER-alpha, ER-beta, coactivators (SRC-1, TIF2, AIB1, CBP, and P/CAF) and corepressors (N-CoR and SMRT) in 6 normal mammary glands, 6 intraductal carcinomas, 22 invasive ductal carcinomas, and 7 breast cancer cell lines using a multiplex reverse transcription-PCR. ER-alpha mRNA expression levels significantly correlated with ER-alpha protein levels measured by enzyme immunoassay in the breast cancer tissues and cell lines. A significant correlation of expression levels was observed between ER-alpha and TIF2, AIB1, P/CAF, and N-CoR, and between ER-beta and AIB1 and CBP in the tissue samples. A significant correlation was also observed between ER-alpha and ER-beta and between ER-beta and CBP in the cell lines. The expression levels of ER-alpha, TIF2, and CBP were significantly higher in the intraductal carcinomas than those in the normal mammary glands. In addition, the expression levels of ER-alpha and N-CoR were significantly higher in the intraductal carcinomas than those in the invasive ductal carcinomas. These findings suggest a positive correlation of expression levels among ER-alpha and cofactors and among ER-beta and cofactors, an up-regulation of expression levels of ER-alpha and cofactors during the development of intraductal carcinomas from normal mammary glands, and a decrease in their expression levels during the progression of breast cancer. PMID- 10690533 TI - Increased expression of cyclooxygenase-2 protein in human gastric carcinoma. AB - Gastric adenocarcinoma is one of the most common malignancies in the world, and yet little is known about its molecular process of development and progression. Recent studies have suggested that ingestion of nonsteroid anti-inflammatory drugs reduces the risk of colon cancer, presumably by inhibiting the cyclooxygenase (COX) enzyme. COX-2, one isoform of the COX enzyme, is the rate limiting enzyme in prostaglandin synthesis, and the function of this enzyme is thought to relate to inflammatory processes and carcinogenesis. To understand the role of COX enzyme in gastric cancer, we measured COX-2 expression in 104 human gastric carcinoma tissues by immunohistochemical analysis. We obtained tissue specimens from 104 surgically resected gastric adenocarcinoma patients. We performed immunohistochemical stain for human COX-2 with polyclonal antibody in gastric carcinoma. After curative resection and extensive lymph node dissection, all patients received adjuvant chemotherapy containing 5-fluorouracil. Expression of COX-2 showed cytoplasmic staining, not only in cancer cells but also in precancerous lesions such as metaplastic and adenomatous cells. We confirmed up regulation of COX-2 in gastric cancer tissues compared with normal paired mucosa using Western blot analysis. There was no correlation between clinicopathological characteristics of gastric cancer patients and intensity of COX-2 protein expression. This study indicates that COX-2 protein over-expression may contribute to an early event of gastric cancer development, and it further suggests that selective inhibition of COX-2 may provide a chemopreventive effect against gastric carcinogenesis. PMID- 10690534 TI - p53 gene mutations are associated with shortened survival in patients with advanced non-small cell lung cancer: an analysis of medically managed patients. AB - Mutations in the p53 gene are common in many cancers. Nevertheless, the relationship between mutations of this tumor suppressor gene and patient survival in non-small cell lung cancer (NSCLC) remains unclear. Interpretation of prior studies of patient outcomes are complicated by the inclusion of both surgical and nonsurgical patients. To better isolate the potential effects of p53 gene mutations per se on tumor progression, we chose to examine patients with advanced disease in whom surgery was not performed (stages IIIA, IIIB, and IV). We have used PCR-denaturing gradient gel electrophoresis, a sensitive and specific method for the detection of a variety of p53 mutations in cytology or biopsy specimens, to evaluate the prognostic significance of p53 gene mutations in nonsurgical patients with advanced NSCLC. In 70 consecutive medical patients, p53 mutations were found in 29 cases (41%) at the time of initial diagnosis. Followed prospectively, patients with p53 mutations had a significantly reduced survival time after diagnosis than those without mutations (median survival, 17 versus 39 weeks; P = 0.0003) independent of other clinical factors. This abbreviated survival occurred in both patients who received chemotherapy (n = 39, P = 0.002) or best supportive care (n = 31, P = 0.018). These results indicate that mutations of the p53 gene in patients with NSCLC who do not undergo surgical resection portends a significantly worse prognosis. PMID- 10690535 TI - Serum tumor marker CA 125 is an early and sensitive indicator of veno-occlusive disease in children undergoing bone marrow transplantation. AB - Veno-occlusive disease (VOD) is a potentially lethal complication of patients undergoing bone marrow transplantation (BMT). The diagnosis of VOD is currently based on clinical signs and unspecific laboratory findings. CA 125 is an oncofetal antigen used as a tumor marker in various malignancies, especially in those originating from the female reproductive tract or gastrointestinal organs, whereas serum CA 125 levels are not increased in hematological malignancies. Several pathophysiological alterations occurring in VOD may lead to elevations in serum CA 125 levels. Therefore, we explored the behavior of this marker as a diagnostic tool in VOD. Twenty-nine pediatric transplant patients were studied. Eight patients (28%) developed clinical VOD, and a significant increase in serum CA 125 was noted in all of them. During the 7 days preceding the diagnosis of VOD, an increase of at least 57% in serum CA 125 from the pre-BMT value was observed in 6 (86%) of 7 of the evaluable patients with VOD. In contrast, a similar increase was noted in only 6 of the 21 non-VOD patients during the post BMT period of 30 days. Accordingly, the sensitivity and specificity of serum CA 125 for predicting or detecting VOD were 86% and 71%, respectively. The serum levels of CA 125 were not affected by the presence of Graft-versus-Host Disease (GvHD) or a septic infection. In conclusion, serum CA 125 is of value as an early marker of VOD in children undergoing BMT. PMID- 10690536 TI - Mutational analysis of the transforming growth factor beta receptor type II gene in hereditary nonpolyposis colorectal cancer and early-onset colorectal cancer patients. AB - Somatic mutations in the transforming growth factor beta receptor type II (TGF beta RII) gene have been observed in various human cancers showing microsatellite instability. Most of the mutations observed were additions or deletions of the mononucleotide repeat sequence present in TGF-beta RII coding region, suggesting that the TGF-beta RII may be a target gene of genomic instability in tumorigenesis. Recently, we reported germ-line frameshift mutations in the mononucleotide repeat sequence of the hMSH6 gene, which is believed to be one of the target genes of genomic instability in tumorigenesis, suggesting the possibility of germ-line mutation in mononucleotide repeat sequences. Moreover, one case of germ-line mutation in the TGF-beta RII gene was identified in a hereditary nonpolyposis colorectal cancer (HNPCC) kindred, indicating the involvement of TGF-beta RII inactivation in tumorigenesis of HNPCC. However, germ line mutation analysis of all of the coding sequences and the mononucleotide repeat sequence of the TGF-beta RII in HNPCC patients has not yet been fully elucidated. Therefore, to further investigate the presence of germ-line mutations, we screened all of the coding region sequences and mononucleotide repeat sequence of TGF-beta RII from 35 HNPCC, 44 suspected HNPCC, and 45 sporadic early-onset colorectal cancer patients. However, no pathogenic mutations other than silent mutations, introgenic mutation, and polymorphisms were identified. Two silent mutations at codons 309 (ACG to ACA) and 340 (CAT to CAC) in the kinase domain located in exon 4 were detected. A 1-bp cytidine deletion was observed 6 bases from the 3' end of intron. Two polymorphisms were identified at codon 389 (AAC to AAT) and at the fourth-to-last base in intron 3. The polymorphism at codon 389 was more frequent in HNPCC (20%; 7 of 35) and suspected HNPCC patients (18%; 8 of 44) than in nonmalignant control group (10%; 5 of 50). Moreover, the frequency was significantly higher in early-onset colorectal cancer patients (31%; 14 of 45). This is the first report of a different frequency of polymorphism in HNPCC, suspected HNPCC, early-onset colorectal cancer patients, and healthy normal individuals. This result suggests that: (a) germ-line mutation of the TGF-beta RII gene may be a rare event during tumorigenesis in HNPCC and sporadic early-onset colorectal cancer; (b) the mononucleotide repeat sequence of the TGF-beta RII gene is an apparent target of genomic instability but not of germ-line mutation; and (c) the polymorphism of codon 389 (AAC to AAT) is frequent, especially in early-onset colorectal cancer patients, in which it is more frequent than in control group. PMID- 10690537 TI - Topological analysis of p21WAF1/CIP1 expression in esophageal squamous dysplasia. AB - In the normal stratified squamous epithelium of the esophagus, only the third to the fifth layers of cells express the cyclin-dependent kinase inhibitor p21WAF1/CIP1 (p21). Using immunohistochemical staining, we examined the topological distribution of cells expressing p21, p53, Ki67, and cytokeratin 10 (CK10), a differentiation marker of esophageal squamous cell carcinoma (SCC), in 25 superficial SCCs and 72 dysplastic lesions of the esophagus. Image analysis of p21, p53, and Ki67 expression was also performed in 48 dysplastic lesions. In superficial SCCs, although Ki67- and p53-expressing cells were mainly distributed in the deep layers of tumors despite tumor differentiation, the distribution of p21 correlated with tumor differentiation. In dysplastic lesions, p53- and Ki67 coexpressing cells tended to locate in the same layers and expand in the lower layers of epithelium with the progression of dysplasia. p21-expressing cells shifted to the upper layers of the epithelium with the progression of dysplasia. However, this change was heterogeneous; in some lesions, p21-expressing cells were confined to the superficial layers of atypical cells (confined type), whereas in others, p21-overexpressing cells were scattered among atypical cells (scattered type). CK10 expression was observed in 25% of dysplastic lesions, and the frequency of CK10 expression was significantly higher in the scattered than in the confined type. Our results suggest that esophageal squamous dysplasia represents the earliest pathological process in esophageal squamous carcinogenesis. Our results also suggest that differentiation of esophageal SCC is determined at the stage of dysplasia, and that p21 plays a critical role in the differentiation process. PMID- 10690538 TI - Expression of protein gene product 9.5 and tyrosine hydroxylase in childhood small round cell tumors. AB - Small round cell tumors of childhood can be histologically ambiguous, can require tumor markers for an accurate diagnosis, and include neuroblastoma, peripheral primitive neuroectodermal tumor (pPNET), Ewing's sarcoma (ES), lymphoma, and rhabdomyosarcoma. Because the cell type of origin for ES remains controversial, characterizing gene expression in ES can provide diagnostic markers and lead to better understanding of tumor biology. We studied RNA expression of the neuronal genes protein gene product 9.5 (PGP 9.5) and tyrosine hydroxylase (TH) by Northern analysis in cell lines and tissue from small round cell tumors. PGP 9.5 showed strong expression in 17 of 17 neuroblastoma cell lines, 9 of 9 pPNET cell lines, and 11 of 11 ES cell lines. PGP 9.5 was weakly expressed in 1 of 1 alveolar rhabdomyosarcoma cell lines but not in 1 of 1 embryonal rhabdomyosarcomas, and weak expression was seen in 1 of 7 leukemia cell lines. In tumor tissue, all 12 neuroblastomas expressed PGP 9.5, as did all 7 pPNET and all 7 ES. PGP 9.5 was very weakly expressed in 6 of 9 rhabdomyosarcomas and 1 of 9 lymphomas. TH was expressed only in neuroblastomas, and no TH expression was seen in cell lines or tissue from other tumors. As high expression of PGP 9.5 was only found in neural tumors; PGP 9.5 expression by ES provides further evidence for a neural origin of this tumor, whereas TH expression is highly specific for neuroblastomas. PGP 9.5 expression should allow sensitive detection of minimal residual disease for ES and pPNET using reverse transcription-PCR, and the variability in TH and PGP 9.5 expression levels in neuroblastomas indicates that expression of both genes should be used for monitoring minimal residual disease by reverse transcription-PCR. PMID- 10690539 TI - Prognostic implication of microsatellite alteration profiles in early-stage non small cell lung cancer. AB - Development of non-small cell lung cancer (NSCLC) is a result of multiple accumulated genetic abnormalities. Profiles of genetic abnormalities may determine tumor behavior and impact on patient outcome. We used microsatellite markers at 3p14, 9p21, and 10q24 to analyze tumor samples from 91 patients with pathologically confirmed stage I NSCLC for microsatellite alterations. Loss of heterozygosity at any single locus was not significantly associated with length of survival. However, patients whose tumors had microsatellite instability (MI) at 10q24 had shortened disease-specific survival. Among 31 such patients, 32% (10 of 31 patients) had died of the disease within 5 years after surgery compared with 16% (9 of 58 patients) without MI at 10q24 (P = 0.07). Interestingly, in the adenocarcinoma subtype, 71% (5 of 7 patients) of the patients with MI at 10q24 succumbed to the disease as compared with only 12% (3 of 26) of the adenocarcinoma patients without such MI (P < 0.001), suggesting the presence of distinct mechanisms in tumorigenesis among different subtypes of lung cancer. It has been noticed that certain microsatellite alteration profiles provide additional values for risk assessment. Of 23 patients who had MI at 10q24 and an alteration at 3p14, 39% (9 of 23 patients) died of the disease within 5 years as compared with only 15% (10 of 66 patients) of the patients without such a profile (P = 0.02). Strikingly, among the 22 patients with no alteration at any loci tested or with loss of heterozygosity at 10q24 and retention of at least one of the other two loci, none died of lung cancer within 5 years after surgery, whereas 28% (19 of 67 patients) of the patients outside these profiles did so (P = 0.01). Our results support the hypothesis that microsatellite alterations can be used as biomarkers for the genetic classification of pathological stage I NSCLC, which may in turn influence treatment decisions dependent on an accurate forecast of patient survival time. PMID- 10690540 TI - Neurotensin receptor-1 mRNA analysis in normal pancreas and pancreatic disease. AB - By autoradiography, neurotensin (NT) binding is specifically detectable in pancreatic cancer, but not in the normal pancreas, chronic pancreatitis (CP), or other pancreatic disorders. In the present study, we investigated whether this is due to NT receptor-1 (NTR-1) mRNA up-regulation and whether NTR-1 mRNA could also be used as a specific diagnostic marker and treatment target in pancreatic cancer. Fifteen normal pancreas tissue samples, 20 CP samples, and 30 pancreatic cancer samples were studied. Expression and localization of NTR-1 mRNA was investigated by Northern blot analysis and in situ hybridization. Furthermore, consecutive tissue sections were analyzed for NTR-1 mRNA expression and NT binding. By Northern blot analysis, NTR-1 mRNA expression was 4.4-fold (P < 0.01) and 3.0-fold (P < 0.01) higher in pancreatic cancer and CP tissue samples, respectively, compared with normal controls. There was no difference in NTR-1 mRNA levels between CP and cancer samples (P > 0.05). In pancreatic cancer, the NTR-1 mRNA levels were higher in advanced tumor stage (stages III and IV) than early tumor stage (stages I and II; P < 0.05), but no difference was found between well/moderately differentiated (grades 1 and 2) and poorly differentiated/undifferentiated cancers (grades 3 and 4; P > 0.05). By in situ hybridization, NTR-1 mRNA signals were weakly present in the cytoplasm of acinar and ductal cells of the normal pancreas. Moderate to intense NTR-1 mRNA signals were present in the cytoplasm of acinar cells dedifferentiating into tubular complexes and degenerating acinar cells of CP samples. In the cancer samples, NTR 1 mRNA was moderately to intensely expressed in the cytoplasm of cancer cells. When on consecutive tissue sections NTR-1 mRNA expression was compared with the presence of NTR-1, measured by receptor autoradiography, a correlation was found in carcinomas but not in CP samples, in which no receptors were detectable by autoradiography. The enhanced expression of NTR-1 mRNA in pancreatic cancer cells further suggests that neuroendocrine hormones might modulate pancreas cancer cell behavior. However, its relatively high levels in CP excludes NTR-1 mRNA as a specific parameter for pancreatic cancer and for the differentiation of pancreatic cancer from CP. PMID- 10690541 TI - Vascular endothelial growth factor expression in untreated osteosarcoma is predictive of pulmonary metastasis and poor prognosis. AB - To investigate the clinical significance of vascular endothelial growth factor (VEGF) in osteosarcoma, we immunohistochemically stained biopsy specimens of 27 primary osteosarcomas using an antibody against VEGF and evaluated the correlation between the expression of VEGF and local density of CD34-positive microvessels, clinicopathological variables, and survival of patients. VEGF staining was positive in 17 tumors (63.0%) in which the density of CD34-positive microvessels was significantly higher than that in VEGF-negative 10 tumors (P < 0.05). In terms of clinicopathological variables, there was no correlation between the expression of VEGF and histological subtype, stage, or response to neoadjuvant chemotherapy, or, strikingly, to the development of pulmonary metastasis (89% of VEGF-positive tumors versus 10% of VEGF-negative tumors; P < 0.0003). Moreover, patients with a VEGF-positive tumor were poorer in both disease-free survival (P < 0.001) and overall survival (P < 0.03) compared to those with a VEGF-negative tumor. These findings strongly suggest that VEGF expression in untreated osteosarcoma is predictive of pulmonary metastasis and poor prognosis in patients who underwent aggressive therapy and also provide the basis for a therapeutic strategy targeting angiogeneic property of osteosarcoma. PMID- 10690542 TI - Cathepsin B, a prognostic indicator in lymph node-negative breast carcinoma patients: comparison with cathepsin D, cathepsin L, and other clinical indicators. AB - New prognosticators are needed for breast cancer patients after the initial surgical treatment to make therapeutic decisions that ultimately will affect their DFS. These consist of specific proteolytic enzymes including lysosomal endopeptidases. In this study, the activity and protein concentrations of cathepsins (Cats) D, B, and L were measured in 282 invasive breast tumor cytosols. These potential biological prognostic indicators were compared with other histopathological parameters, such as tumor size, lymph node involvement, tumor-node-metastasis stage, histological grade, DNA analysis, and steroid receptors. CatD protein concentration correlated with lymph node involvement. CatB and CatL levels correlated significantly with Scarf-Bloom-Richardson histological grade and were also higher in estrogen-negative tumors, and CatB was higher in larger tumors. As prognostic markers, CatB concentration was significant for increased risk for recurrence in the entire patient population and specifically also in lymph node-negative patients as follows: high CatB concentration (above 371 micrograms/g) in tumor cytosols was significant (P < 0.00) for high risk of recurrence but was of only borderline prognostic significance (P < 0.06) for overall survival of all patients. In lymph node negative patients, CatB (above 240 micrograms/g, P < 0.003) was highly significant for recurrence-free survival, followed by CatL (above 20 micrograms/g, P < 0.049) and CatD (above 45 nmol/g, P < 0.044) concentrations. For overall survival of node-negative patients, only CatB was a significant (P < 0.014) prognosticator. We conclude that CatB is useful as a prognostic indicator in lymph node-negative patients. This suggests that selective adjuvant therapy should be applied in this lower risk group of patients when high levels of CatB are determined. PMID- 10690543 TI - In multiple myeloma, circulating hyperdiploid B cells have clonotypic immunoglobulin heavy chain rearrangements and may mediate spread of disease. AB - DNA aneuploidy characterizes a proportion of malignant bone marrow (BM)-localized plasma cells in multiple myeloma (MM). This analysis shows that for most MM patients, circulating clonotypic B cells in MM are also hyperdiploid. Although all normal B cells and some malignant B cells are diploid, hyperdiploidy is likely to be exclusive to those that are malignant. Hyperdiploid MM B cells express CD34 and have clonotypic IgH transcripts, confirming them as part of the malignant clone. For MM, 92% (70/76) of patients had a DNA hyperdiploid subset [5 30% of peripheral blood mononuclear cells (PBMCs)] of CD19+ B cells. All CD19+ PBMCs in MM expressed CD19 and IgH variable diversity joining (VDJ) transcripts, confirming them as B cells. DNA aneuploid cells were undetectable in T or B lymphocytes from normal blood, spleen or thymus, or in blood from patients with B chronic lymphocytic leukemia. In MM, untreated patients had the highest DNA index (1.12). DNA hyperdiploid PBMCs were most frequent among untreated patients and were significantly reduced after chemotherapy. Diploid B cells were significantly more frequent after chemotherapy than at diagnosis. Of the hyperdiploid PBMCs, 81 +/- 3% expressed CD34 and CD19. In contrast to circulating CD34+ B cells, CD34- B cells in MM are diploid. In MM, unlike hyperdiploid PBMC B cells, hyperdiploid BM plasma cells lack both CD34 and CD19, suggesting that loss of CD34 correlates with differentiation and BM anchoring. In situ reverse transcription-PCR of the CD34+ (hyperdiploid) and CD34- (diploid) PBMC B-cell subsets was performed using patient-specific primers to amplify clonotypic IgH VDJ transcripts. Confirming previous work, CD34+ hyperdiploid MM PBMCs were clonotypic (86 +/- 5%). In contrast, CD34- diploid MM PBMCs had few monoclonal cells (4.8 +/- 2%). The lack of hyperdiploidy, together with the relative absence of cells having clonotypic transcripts, suggests these polyclonal CD34- B cells are normal. After culture in colchicine to arrest mitosis, hyperdiploid B cells were reduced and MM B cells accumulated in a diploid G2-M, suggesting that hyperdiploid in MM may represent a transient S-phase arrest rather than an aneuploid G0 phase. The DNA hyperdiploidy of CD34+ clonotypic B cells suggests these cells may be clinically important constituents of the myeloma clone and that they may play a direct role in the spread of myeloma. PMID- 10690544 TI - Significance of neuron-specific enolase levels before and during therapy for small cell lung cancer. AB - The level of serum neuron-specific enolase (NSE) has been implicated as a prognostic factor for patients with small cell lung cancer (SCLC). A prospective evaluation was undertaken to assess the prognostic significance of pretreatment NSE and treatment-induced minimum NSE values in patients with SCLC. Patients from two Phase III North Central Cancer Treatment Group trials [one for patients with extensive stage SCLC and one for patients with limited stage SCLC] were asked to enter this laboratory correlational trial. Both trials included treatment with four to six cycles of etoposide and cisplatin, and 121 patients (71 extensive stage SCLC and 50 limited stage SCLC) were entered into the present study of NSE. Pretreatment NSE values and treatment-induced minimum NSE values were independent predictors of time to progression and survival in multivariate analysis. Hazard rate modeling allowed the formulation of specific relationships of NSE to time to progression and survival. Pretreatment NSE levels inversely correlated with time to progression and survival in these patients with SCLC. Pretreatment NSE accounted for 28% of the variance in survival. Both pretreatment NSE and treatment-induced minimum NSE were independent prognostic predictors of time to progression and survival. PMID- 10690545 TI - Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. AB - The DNA base excision repair pathway is responsible for the repair of alkylation and oxidative DNA damage. A crucial step in the base excision repair pathway involves the cleavage of an apurinic/apyrimidinic (AP) site in DNA by an AP endonuclease (APE). The major AP endonuclease in mammalian cells is APE/ref-1, a multifunctional enzyme that acts not only as an AP endonuclease but as a redox modifying factor for a variety of transcription factors. The purpose of this study was to determine the expression of APE/redox factor-1 (ref-1) in ovarian tissues, particularly ovarian cancers. Formalin-fixed, paraffin-embedded specimens of ovarian tissues (normal, various benign conditions, and epithelial cancers) were studied using both polyclonal and monoclonal antibodies to APE/ref 1. The relationship between APE/ref-1 protein levels and DNA repair activity was studied in ovarian Hey and Hey-C2 cell lines using Western blot and a specific AP site oligonucleotide cleavage assay. Hey and Hey-C2 cells were fractionated, and the nuclear and cytoplasmic extracts were quantitated for protein levels and assessed for APE/ref-1 with Western blot. Normal ovarian tissues consistently demonstrated strong nuclear staining of the surface epithelium, epithelial inclusions, corpora lutea and albicantia, and stroma. Cytoplasmic staining was absent. A similar pattern was seen for benign conditions including endometriosis. Low malignant potential ovarian cancers stained in a pattern similar to normal ovarian and nonneoplastic tissues; however, two specimens also had areas of cytoplasmic staining. Epithelial ovarian cancers were remarkably different from all other ovarian tissues studied. Both nuclear and cytoplasmic staining of the malignant epithelium were seen and ranged from strong to weak, often with considerable staining heterogeneity within the same tumor. The AP-site oligonucleotide cleavage assay indicated that APE/ref-1 protein levels correlate well with DNA repair activity. The increased levels of APE/ref-1 in the Hey-C2 cells was mainly attributable to increased cytoplasmic enzyme. APE/ref-1 immunoreactivity is altered in malignant ovarian tumors. Further studies will determine whether the altered expression and subcellular location reflect changes in redox regulatory functions. PMID- 10690546 TI - Bladder cancer: allelic deletions at and around the retinoblastoma tumor suppressor gene in relation to stage and grade. AB - Inhibition of the retinoblastoma tumor suppressor gene (RB) is probably important in the pathogenesis of urinary bladder cancer. Little information is available concerning allelic loss on 13q11 to 13q32 and its relation to grade and stage. In a population-based study, freshly frozen tissue was collected from all new cases of urinary bladder cancer in the Stockholm region during 1995-1996. Here we report the occurrence of loss of heterozygosity (LOH) at seven sites in 13q11 to 13q32 as analyzed in 236 cases by a fluorescent multiplex PCR-based on tumor DNA and peripheral blood. For each site, about 30% of the cases were not informative because of homozygosity. Replication errors were detected in 4% (17 cases). LOH was found in 21 (at 13q11-12.1) to 32% (at 13q14.3 in RB) of the informative cases. A correlation was found between the prevalence of LOH at all observed loci and stage and grade, respectively, and it was statistically significant for 13q14.3. LOH at RB was found in Ta as well as grade 1 tumors. Also, a statistically significant correlation was found between the number of loci with LOH at 13q and tumor stage and grade, respectively. Typically an altered RB function is related to the expected clinical course of urinary bladder cancer, but allelic loss including the gene also occurs in low grade and low stage tumors. An altered RB function probably is not necessary for a malignant transformation of urothelial cells. The causal direction of the relation between the quantity of the deleted DNA and tumor aggressiveness is not clear. PMID- 10690547 TI - Prediction of clinical outcome from primary tamoxifen by expression of biologic markers in breast cancer patients. AB - The aim of this study was to evaluate pretreatment clinical features and biological markers together with changes in these factors as predictors of response and relapse in patients receiving tamoxifen for primary breast cancer. Fine-needle aspiration cytology of the primary breast cancer was performed before tamoxifen treatment in 54 patients and repeated after therapy on day 14, day 60, or on both days in a subset of 35 patients. These samples were evaluated for estrogen receptor (ER), progesterone receptor (PgR), Ki67, S-phase fraction and ploidy. The overall response to tamoxifen was 57% (31 of 54 patients). Pretreatment ER and PgR significantly predicted for response by univariate analysis (P < 0.0001 and P < 0.003, respectively). By multivariate analysis, ER expression was the only independent predictor of response, and it was associated with 27 times the likelihood of response (95% confidence interval, 6-136). Increase in PgR and decrease in Ki67 on day 14 significantly predicted for response to tamoxifen (P < 0.03 and P < 0.04, respectively). Lack of ER, clinical node-positive disease, and failure to decrease Ki67 on day 14 were significantly associated with increased risk of relapse (P < 0.05). By multivariate analysis, ER expression was the only independent predictor of relapse (P < 0.005). Pretreatment and early changes in molecular marker expression may assist in the prediction of response and clinical outcome in primary breast cancer patients receiving tamoxifen. PMID- 10690548 TI - Enhancement of angiogenesis, tumor growth, and metastasis by transfection of vascular endothelial growth factor into LoVo human colon cancer cell line. AB - The expression of vascular endothelial growth factor (VEGF), a highly potent angiogenic molecule, is thought to be correlated with the development of colon cancer; however, direct evidence for a role of VEGF in metastasis is lacking. This study was designed to more directly establish the role of VEGF in the growth and metastasis of human colon cancer using a genetically engineered cancer cell line. A stable VEGF gene-transfected human colon cancer cell line, LoVo, was made by genetic manipulation using eukaryotic expression constructs designed to express the complete VEGF121 cDNA in the sense orientation. Transfected clones were screened for VEGF121 mRNA expression by Northern blot analysis and for VEGF121 protein expression by Western blot analysis. Consequently, we obtained S17 cells that expressed a high level of both VEGF mRNA and VEGF protein. A vector-transfected clone (V7 cell) was used as a control. The experiment with the dorsal air sac method revealed that S17 cells elicited a stronger directional out growth of capillaries than V7 cells. S17 cells formed faster-growing tumors than did V7 cells when xenografted s.c. into nude mice, although there was no significant difference in their in vitro proliferation. Tumors derived from S17 cells had more vascularity, as assessed by counting capillary vessels after staining with factor VIII, than did tumors derived from V7 cells (P < 0.05). With regard to the metastatic potential, S17 cells exhibited a higher capacity for both hepatic metastasis after the splenic portal inoculation and peritoneal dissemination after i.p. injection than did V7 cells. However, S17 cells showed no apparent metastasis, despite their rapid growth after orthotopic implantation. In conclusion, the present study showed clearly that VEGF plays an important role in cancer growth due to stimulation of angiogenesis by accelerating cell growth after reaching the target organs. PMID- 10690549 TI - The combined use of an immunotoxin and a radioimmunoconjugate to treat disseminated human B-cell lymphoma in immunodeficient mice. AB - Immunoconjugates (ICs) consist of a targeting moiety and a toxic moiety and have the specificity that traditional cancer therapy lacks. At appropriate doses, ICs are safe and effective in treating various cancers in experimental animals and in humans. However, because cures are rarely achieved using single agents, regimens involving combinations of agents with different mechanisms of action must be evaluated. In this study, we explored the efficacy and toxicity of a combination of two IC therapies, radioimmunotherapy (RIT) and immunotoxin (IT) therapy, to treat advanced, disseminated human lymphoma in immunodeficient mice. We proposed to use the bystander effect of RIT to reduce large tumor burdens, followed by an IT to eliminate residual tumor cells. Our results indicate that, when used alone, both RIT and IT therapy were safe and effective, but not curative. When the two therapies were combined, efficacy and toxicity became dependent on the temporal order of administration. Thus, with the doses used in this study, when RIT was administered after IT therapy, the regimen was curative. In contrast, when RIT was administered before IT therapy, the combination was highly toxic or even lethal. Both RIT and IT therapy induced pulmonary vascular leak, but with different kinetics. When RIT was given prior to IT therapy, the pulmonary vascular leak became life-threatening but not when the two agents were administered in the reverse order. PMID- 10690550 TI - Methionine depletion enhances the antitumoral efficacy of cytotoxic agents in drug-resistant human tumor xenografts. AB - Efficacy of chemotherapy is limited in numerous tumors by specific cellular mechanisms that inactivate cytotoxic antitumoral drugs, such as ATP-dependent drug efflux and/or drug detoxification by glutathione. In reducing ATP pools and/or glutathione synthesis, it might be possible to enhance the efficacy of drugs affected by such resistance mechanisms. Reduction of the ATP pool and glutathione content is achievable in cancer cells by depleting the exogenous methionine (Met) supply and ethionine. Thus, the rationale for the present study was to use Met depletion to decrease the ATP and glutathione pools so as to sensitize tumors refractory to cytotoxic anticancer drugs. Met depletion was achieved by feeding mice a methionine-free diet supplemented with homocysteine. The effects of Met depletion combined with ethionine and/or chemotherapeutic agents were studied using human solid cancers xenografted into nude mice. TC71-MA (a colon cancer) SCLC6 (a small cell lung cancer), and SNB19 (a glioma) were found to be refractory to cisplatin, doxorubicin, and carmustine, respectively. These three drugs are used to treat such tumors and are dependent for their activity on the lack of cellular ATP- or glutathione-dependent mechanisms of resistance. TC71-MA, SCLC6, and SNB19 were Met dependent because their proliferation in vitro and growth in vivo were reduced by Met depletion. Cisplatin was inactive in the treatment of TC71-MA colon cancer, whereas a methionine-free diet, alone or in combination with ethionine, prolonged the survival of mice by 2-fold and 2.8-fold, respectively. When all three approaches were combined, survival was prolonged by 3.3-fold. Doxorubicin did not affect the growth of SCLC6, a MDR1-MRP-expressing tumor. A Met-deprived diet and ethionine slightly decreased SCLC6 growth and, in combination with doxorubicin, an inhibition of 51% was obtained, with survival prolonged by 1.7-fold. Combined treatment produced greater tumor growth inhibition (74%) in SCLC6-Dox, a SCLC6 tumor pretreated with doxorubicin. Growth of SNB19 glioma was not inhibited by carmustine, but when it was combined with Met depletion, survival duration was prolonged by 2-fold, with a growth inhibition of 80%. These results indicate the potential of Met depletion to enhance the antitumoral effects of chemotherapeutic agents on drug-refractory tumors. PMID- 10690551 TI - Prolonged response to antisense cyclin D1 in a human squamous cancer xenograft model. AB - Local recurrence of squamous cell cancer (SCC) causes high morbidity and is often readily accessible, making such patients potential candidates for gene therapy. Cyclin D1 (CD1), critical in the G1-S transition in the cell cycle, is amplified in 20-50% and overexpressed in up to 80% of head and neck SCC. Our earlier studies indicated that CD1 expression increased with progression from low grade to high grade dysplasia, and that treatment of established tumors with antisense cyclin D1 (AS-cyclin D1) led to tumor regression during a one week evaluation period. We hypothesized that: 1) CD1 expression increases with disease progression to advanced SCC, and 2) AS-cyclin D1 therapy would lead to prolonged tumor regression in a xenograft model of human SCC. CD1 expression, evaluated by immunostain in 30 stage III/IV head and neck SCC, increased in the basal layer from normal-dysplasia (P = 0.06) and from dysplasia-carcinoma (P = 0.004). In the germinative layer CD1 expression increased from dysplasia-carcinoma (P = 0.002) but not from normal-dysplasia. Western blotting of eight SCC and two transformed keratinocyte cell lines demonstrated CD1 overexpression in 8/10 (80%) lines. An 11th cell line (A431) had previously been shown to overexpress cyclin D1. 8/9 (89%) cell lines overexpressing CD1 formed tumors in immunodeficient mice, whereas 0/2 cell lines without CD1 overexpression formed a tumor. Three established SCCs, one fast growing, one with moderate growth rate (with CD1 overexpression) and one slow growing (without increased CD1), shrank significantly for 2-4 weeks after AS-cyclin D1 treatment, while tumors transduced with control vector grew. Cyclin D1 expression increases in frequency with disease progression, and antisense cyclin D1 was effective in a xenograft model of human cancer, independent of tumor growth rate. PMID- 10690552 TI - Characterization of an ovarian carcinoma cell line resistant to cisplatin and flavopiridol. AB - Flavopiridol, the first inhibitor of cyclin-dependent kinases to enter clinical trials, has shown promising antineoplastic activity and is currently undergoing Phase II testing. Little is known about mechanisms of resistance to this agent. In the present study, we have characterized an ovarian carcinoma cell line [OV202 high passage (hp)] that spontaneously developed drug resistance upon prolonged passage in tissue culture. Standard cytogenetic analysis and spectral karyotyping revealed that OV202 hp and the parental low passage line OV202 shared several marker chromosomes, confirming the relatedness of these cell lines. Immunoblotting demonstrated that OV202 and OV202 hp contained similar levels of a variety of polypeptides involved in cell cycle regulation, including cyclin dependent kinases 2 and 4; cyclins A, D1, and E; and proliferating cell nuclear antigen. Despite these similarities, OV202 hp was resistant to flavopiridol and cisplatin, with increases of 5- and 3-fold, respectively, in the mean drug concentrations required to inhibit colony formation by 90%. In contrast, OV202 hp and OV202 displayed indistinguishable sensitivities to oxaliplatin, paclitaxel, topotecan, 1,3-bis(2-chloroethyl)-1-nitrosourea, etoposide, doxorubicin, vincristine, and 5-fluorouracil, suggesting that the spontaneously acquired resistance was not attributable to altered P-glycoprotein levels or a general failure to engage the cell death machinery. After incubation with cisplatin, whole cell platinum and platinum-DNA adducts measured using mass spectrometry were lower in OV202 hp cells than OV202 cells. Similarly, after flavopiridol exposure, whole cell flavopiridol concentrations measured by a newly developed high performance liquid chromatography assay were lower in OV202 hp cells. These data are consistent with the hypothesis that acquisition of spontaneous resistance to flavopiridol and cisplatin in OV202 hp cells is due, at least in part, to reduced accumulation of the respective drugs. These observations not only provide the first characterization of a flavopiridol-resistant cell line but also raise the possibility that alterations in drug accumulation might be important in determining sensitivity to this agent. PMID- 10690553 TI - Mismatch repair and p53 independently affect sensitivity to N-(2-chloroethyl)-N' cyclohexyl-N-nitrosourea. AB - The contributions of defective mismatch repair (MMR) and the p53-response to cell killing by N-(2-chloroethyl)-N'-cyclohexyl-N-nitrosourea (CCNU) were evaluated. MMR defects were previously shown to be associated with CCNU sensitivity (G. Aquilina et al., Cancer Res., 58: 135-141, 1998). Unexpectedly, eight MMR deficient variants of the A2780 human ovarian carcinoma cell line were 3-fold more resistant to CCNU than the MMR-proficient parental cells. The variants were members of a preexisting subpopulation of drug-resistant A2780 cells. In addition to deficient expression of the MMR protein hMLH1, an essential component of the hMutL alpha repair complex, the variants exhibited alterations in the expression of other genes that influence drug sensitivity. Although A2780 cells possess a wild-type p53 gene, all of the clones contained a heterozygous G to T tranversion at codon 172. This change resulted in a Val to Phe substitution and was associated with a constitutive production of high levels of p53, which was inactive as a transcriptional activator of bax and p21. The hMLH1/p53 defective variants displayed a less prominent cell cycle arrest and reduced apoptosis after CCNU treatment. In contrast, MMR-defective A2780 variants, which had a similar hMutL alpha defect but retained a wild-type p53, did exhibit the expected CCNU sensitivity. Expression of a dominant-negative p53val135 increased CCNU resistance of both MMR-proficient and MMR-deficient A2780 cells. Thus, defective MMR and p53 influence CCNU sensitivity in opposite directions. Their effects are independent, and sensitization by defective MMR does not require a functional p53 response. PMID- 10690554 TI - Induction of apoptosis in malignant B cells by phenylbutyrate or phenylacetate in combination with chemotherapeutic agents. AB - Phenylacetate (PA) and phenylbutyrate (PB) are aromatic fatty acids that are presently undergoing evaluation as potential antineoplastic agents. In vitro, PA and PB cause differentiation or growth inhibition of malignant cells. Clinical trials of these drugs as single agents indicate that they are not myelosuppressive; therefore, combinations with other chemotherapy agents may be possible. The goals of this study were to determine whether PA and PB (a) are cytotoxic to malignant B cells from patients with non-Hodgkin's lymphoma and B cell chronic lymphocytic leukemia and (b) exhibit additive or synergistic induction of apoptosis when administered to myeloma cell lines in combination with conventional drugs. In the clinical specimens, cytotoxicity was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and percent apoptosis was measured using 7-aminoactinomycin D and flow cytometry. Viability was decreased by > 50% in 7% (1/15) of non-Hodgkin's lymphoma samples treated with 5 mM PA, 27% treated with 1 mM PB, and 60% treated with 2 mM PB. Likewise, viability was decreased by > 50% in 44% (4/9) of chronic lymphocytic leukemia samples treated with 5 mM PA, 67% treated with 1 mM PB, and 100% treated with 2 mM PB. Studies in the myeloma cell lines demonstrated that PB treatment induced activation of caspases 3, 7, and 9 accompanied by cleavage of their substrates and internucleosomal DNA degradation. Combinations of PA or PB with conventional drugs (cytarabine, topotecan, doxorubicin, etoposide, chlorambucil, melphalan, fludarabine, carboplatin, and cisplatin) were examined for synergism (combination index < 1 in median effect analysis) in inducing apoptosis of both the MY5 and 8226 human myeloma cell lines. At concentrations that killed > 50% of cells, most combinations were additive; however, PB was synergistic with cytarabine, etoposide, and topotecan, with the combination index < 1 at each of the 50, 75, and 95% apoptosis levels. These observations indicate that PA and PB can induce apoptosis in malignant B cells and enhance the cytotoxicity of agents used in the treatment of these malignancies. PMID- 10690555 TI - Hairy cell leukemia, a B-cell neoplasm that is particularly sensitive to the cytotoxic effect of anti-Tac(Fv)-PE38 (LMB-2). AB - Anti-Tac(Fv)-PE38 (LMB-2) is a recombinant, single-chain immunotoxin composed of the variable domains of the anti-Tac (anti-CD25) monoclonal antibody fused to a truncated form of Pseudomonas exotoxin (PE). Until now, this agent has been reported to be very cytotoxic toward T-cell but not B-cell leukemic cells freshly obtained from patients and is being tested clinically in patients with CD25+ malignancies of both B- and T-cell origin. Hairy cell leukemia (HCL) is a B-cell malignancy in which the cells are usually CD25+ and their ex vivo sensitivity to LMB-2 was unknown. Malignant cells from the first HCL patient to be tested were very sensitive to the cytotoxic effect of LMB-2 in vitro (IC50, 1.1 ng/ml), and this patient responded clinically to LMB-2 administered systemically. Therefore, we decided to assess the potential clinical utility of LMB-2 in other patients with HCL. We tested fresh leukemic cells from nine additional CD25+ HCL patients. LMB-2 was very cytotoxic ex vivo in all patients with IC50s as low as 0.5 ng/ml. Malignant cells freshly obtained from patients with chronic lymphocytic leukemia were also sensitive to LMB-2 but not as sensitive as cells from HCL patients. These results indicate that CD25+ HCL is a B-cell neoplasm that is particularly sensitive to LMB-2, and this agent may be useful in patients who have failed standard therapies. PMID- 10690556 TI - In vivo enhancement of tumor radioresponse by C225 antiepidermal growth factor receptor antibody. AB - Overexpression of epidermal growth factor receptor (EGFR) has been correlated with tumor resistance to cytotoxic agents, including radiation (T. Akimoto et al., Clin. Cancer Res., 5: 2884-2890, 1999), and thus is a candidate target for anticancer treatment. This study investigated whether treatment with C225 anti EGFR antibody would improve tumor response to radiotherapy. Nude mice bearing 8 mm-diameter A431 tumor xenografts in the hind leg were treated with C225 antibody, 18 Gy of single-dose local tumor irradiation, or both. C225 was given i.p. at a dose of 1 mg/mouse 6 h before irradiation or 6 h before and 3 and 6 days after irradiation. Delay in tumor growth was the treatment end point. C225 dramatically improved the efficacy of local tumor irradiation, particularly when multiple injections of C225 were administered. Tumor radioresponse was enhanced by a factor of 1.59 by a single dose and by a factor of 3.62 by a doses of C225. Histological analyses of tumors revealed that C225 caused a striking increase in central tumor necrosis associated with hemorrhage and vascular thrombosis when combined with radiotherapy. In addition, C225 induced heavy tumor infiltration with granulocytes, increased tumor cell terminal differentiation, and inhibited tumor angiogenesis. We conclude that C225 anti-EGFR antibody enhances tumor radioresponse by multiple mechanisms that may involve direct and indirect actions on tumor cell survival. PMID- 10690557 TI - Regression of U-87 MG human glioblastomas in nude mice after treatment with a cytotoxic somatostatin analog AN-238. AB - Receptors for somatostatin (SST) found on brain tumors could be used for targeting of chemotherapeutic agents. This study was conducted to investigate the effects of targeted cytotoxic SST analogue AN-238, consisting of 2 pyrrolinodoxorubicin (AN-201), a potent derivative of doxorubicin (DOX) linked to somatostatin analogue RC-121, on the growth of SST receptor-positive U-87 MG human glioblastomas. Nude mice bearing U-87 MG xenografts received i.v. saline or equimolar doses of AN-238 or AN-201 (150 nmol/kg). Experiments also included groups that were administered RC-121 prior to the injection of AN-238, and groups injected with AN-162, a cytotoxic SST analogue similar to AN-238 but containing DOX instead of AN-201. Tumor volume, weight, and burden were determined. The effect of AN-238 and AN-201 on the survival time of nude mice bearing orthotopically implanted U-87 MG tumors was also evaluated. The binding of AN-238 to U-87 MG tumors was determined by radioreceptor assay and SST receptor (SSTR) subtype by reverse transcription-PCR. Nineteen days after a single administration of AN-238 the growth of U-87 MG tumors in nude mice was significantly inhibited (P = 0.00168), whereas two injections of AN-238 produced the regression of tumors (P = 0.0046). AN-201 was toxic and ineffective at the same dose. The antitumor effect on AN-238 could be blocked by pretreatment of the tumor-bearing mice with RC-121. The mean survival time of nude mice inoculated orthotopically with U-87 MG cells into the brain was significantly prolonged by treatment with AN-238 (P = 0.0099). AN-162 failed to inhibit significantly the growth of U-87 MG xenografts. High affinity binding sites for SST and mRNA for SST-2 receptor subtype were detected in U-87 MG tumors. Cytotoxic SST analogue AN-238 can be targeted to SST receptors on U-87 MG human glioblastomas to produce powerful inhibition of growth. PMID- 10690558 TI - Overexpression of Bax enhances antitumor activity of chemotherapeutic agents in human head and neck squamous cell carcinoma. AB - Overexpression of the Bax protein in human head and neck squamous cell carcinoma A253 cells was reported to result in an increased sensitivity to various chemotherapeutic agents in vitro (Guo et al., Oncol. Res., 11: 91-99, 1999). In the present study, the relationship between Bax expression and response to chemotherapy was further investigated in vitro and in vivo model systems. For in vitro study, A253, A253/Vec (pcDNA3 vector transfectant), and A253/Bax (pcDNA3/Bax transfectant, expressing 50-fold higher Bax protein than A253 and A253/Vec) cells were exposed to various concentrations of raltitrexed (a specific thymidylate synthase inhibitor) and SN-38 (a topoisomerase I inhibitor) for 2 h, and cell growth inhibition was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide clonogenic assay. Compared to A253/Vec, A253/Bax cells exhibited 9.5- and 13.5-fold increases in sensitivity to raltitrexed and SN 38, respectively. For in vivo study, A253/Vec and A253/Bax tumor xenografts were established by s.c. injection of tumor cells into nude mice. The antitumor activity and toxicity of raltitrexed (i.v. push daily for 5 days) and irinotecan (a prodrug of SN-38; i.v. push daily for 3 days) were evaluated. The maximum tolerated doses of raltitrexed and irinotecan were 30 and 100 mg/kg/day, respectively. At the maximum tolerated doses, minimal antitumor activity was observed with raltitrexed, although irinotecan was more active than raltitrexed against A253 or A253/Vec tumors. In contrast, both raltitrexed and irinotecan were significantly more active against A253/Bax xenografts than against A253/Vec xenografts; the yield for complete tumor regression (cure) was 40% and 100% with raltitrexed and irinotecan, respectively, with no significant toxicity. Furthermore, the observed increase of antitumor activity in A253/Bax tumors was associated with an enhanced induction of apoptosis in vivo. The in vivo results demonstrated a proof of the principal concept that selecting up-regulation of the proapoptosis gene Bax can provide the basis for a greater therapeutic efficacy to a variety of chemotherapeutic agents with different structures and mechanisms of action. PMID- 10690559 TI - Treatment of isografted 9L rat brain tumors with beta-5-o-carboranyl-2' deoxyuridine neutron capture therapy. AB - beta-5-o-Carboranyl-2'-deoxyuridine (D-CDU) is a nontoxic pyrimidine nucleoside analogue designed for boron neutron capture therapy of brain tumors. In vitro studies indicated that D-CDU accumulates to levels 92- and 117-fold higher than the extracellular concentration in rat 9L and human U-251 glioma cells, respectively, and persists for several hours at levels 5-fold higher than the extracellular concentration. Furthermore, D-CDU was not toxic to rats injected i.p. with up to 150 mg/kg. On the basis of these studies, D-CDU was evaluated as a neutron capture therapy agent using rats bearing stereotactically implanted intracranial 9L tumors at single i.p. doses of 30 mg/kg and 150 mg/kg of D-CDU (20% 10B enriched), given 2 h before irradiation with thermal neutrons. Boron concentrations in tumors 2 h after dosing were 2.3 +/- 1.6 and 7.4 +/- 1.3 micrograms boron/g tissue (mean +/- SD), corresponding to tumor/brain ratios of 11.5 +/- 3.6 and 6.8 +/- 2.0 micrograms boron/g tissue for the low and high doses, respectively. All untreated animals died within 28 days, whereas half survived at days 32, 55, and 38 for groups receiving neutrons only, 30 mg/kg D CDU, and 150 mg/kg D-CDU, respectively. Odds ratios of all treatment groups differed significantly from the untreated group (P < 0.002; logrank test). The median survival time for the 30 mg/kg-treated group but not for the 150 mg/kg treated group was significantly longer than for rats treated with neutrons only (P = 0.036), which may correlate with the decreased tumor selectivity for D-CDU observed at the higher dose. Additional pharmacodynamic studies are warranted to determine optimal dosing strategies for D-CDU. PMID- 10690560 TI - The topoisomerase I inhibitor DX-8951f is active in a severe combined immunodeficient mouse model of human acute myelogenous leukemia. AB - The severe combined immunodeficient (SCID) mouse model of human acute myelogenous leukemia (AML) is a unique system for preclinical in vivo evaluation of the activity and toxicity of new agents. The topoisomerase I (topo I) inhibitor topotecan is active in patients with AML and myelodysplastic syndromes. DX-8951f is a novel topo I inhibitor with more potent antitumor effects than topotecan or CPT-11 in vitro. To study the in vivo activity of DX-8951f, 6-week-old female SCID mice received injections into the tail vein with 2 x 10(7) exponentially growing KBM-3 cells. In each experiment, three to five sets of five mice were treated with DX-8951f doses ranging from 7.5 to 80 mg/kg and at schedules of 1, 3, and 5 days; a control set of five mice was treated with the drug vehicle alone. One group received DX-8951f on day 7 of the inoculation with KBM-3 (early treatment group). To study the activity of DX-8951f in advanced disease, a second group was treated 1 month after the inoculation, when the animals were developing symptoms (late-treatment group). The study end point was the duration of survival until death from leukemia, which was assessed clinically and by the presence of the human DQ alpha gene in tissue samples by PCR. Six experiments were conducted with 170 animals. Survival was higher in both the early- and late-treatment groups than in untreated controls, and the treated groups had significantly less central nervous system disease. Significantly improved survival was observed in animals treated early with 60 and 80 mg/kg as a single injection, with 15 and 20 mg/kg over 3 days, and with 7.5 and 10 mg/kg over 5 days. In the late-disease model (treatment starting on days 28-35), improved survival was observed with a single dose of 80 or 20 mg/kg over 5 days. Dose escalation was limited by dilution problems at the 1-day schedule and by toxicity (mainly gastrointestinal) of the prolonged schedules. Both efficacy and toxicity were dose schedule dependent, increasing with higher doses and prolonged exposure. By establishing the antileukemic activity of DX-8951f against human AML transplanted into SCID mice at doses below the LD10, our data provide a rationale for clinical evaluation of the drug in patients with AML and favor the use of prolonged administration. PMID- 10690562 TI - Glucose homeostasis in the micropremie. AB - This article evaluates the current knowledge of the kinetics of glucose homeostasis in the micropremie. Glucose production, glucose use, and glucose oxidation are reviewed in detail. This article also evaluates the developmental regulation of glucose homeostasis relative to some of the fundamental differences known to exist in the neonate compared to the adult. PMID- 10690561 TI - In vitro and in vivo activity of protein kinase C inhibitor chelerythrine chloride induces tumor cell toxicity and growth delay in vivo. AB - Although clonogenic or divisional death is the main mechanism by which DNA damaging agents demonstrate antitumor activity, recent data indicate that strategies specifically designed to trigger apoptosis may also prove to be useful antitumor agents. Protein kinase C (PKC) isoenzymes are involved in the regulation of cell proliferation, differentiation, and survival. Whereas pharmacological inhibition of PKC activity triggers apoptosis in most mammalian cells, cell line and tissue differences in sensitivities to these inhibitors remain. Whereas PKC inhibitors have potential as antitumor agents, issue of kinase specificity and solubility have remained obstacles to their clinical use. In this report, we investigated the antitumor activity of the PKC inhibitor chelerythrine chloride (chelerythrine), a selective inhibitor of group A and B PKC isoforms. Chelerythrine exhibited cytotoxic activity against nine human tumor cell lines tested in vitro. On the basis of the finding that radioresistant and chemoresistant squamous cell carcinoma lines (HNSCC) undergo apoptosis rapidly after treatment with chelerythrine in vitro, we assessed the effects of this agent on p53-deficient SQ-20B HNSCC cells in vivo. The results demonstrate that chelerythrine treatment of nude mice bearing SQ-20B is associated with significant tumor growth delay. Significantly, treatment with chelerythrine resulted in minimal toxicity. These findings demonstrate a potential for chelerythrine as an antitumor drug against squamous cell carcinoma. PMID- 10690563 TI - Protein metabolism in the extremely low-birth weight infant. AB - Although extensive data are available on the impact of nutrient and protein administration on growth, plasma amino acids, and nitrogen balance in the newborn and growing infants, relatively few studies have carefully examined the dynamic aspects of protein metabolism in vivo and particularly in the micropremie or ELBW infant. These studies show that the very preterm infants, either because of immaturity or because of the intercurrent illness, have high rates of protein turnover and protein breakdown. This high rate of proteolysis is not as responsive to nutrient administration. Intervention strategies aimed at promoting nitrogen accretion, such as insulin, human growth hormone, or glutamine, have not thus far resulted in enhanced protein accretion and growth. This may be, in part, due to limitations in delivery of adequate calorie and nitrogen. PMID- 10690564 TI - Lipid metabolism of the micropremie. AB - Intravenous lipid emulsions often provide substance for the very low-birth weight or extremely low-birth weight infant that need total parenteral nutrition. The process used in this type of treatment as well as the effects of such treatment are discussed at length in this article. Some of the main compounds of representative lipid emulsions are listed and evaluated and the benefits and consequences of their use are presented. PMID- 10690565 TI - Essential fatty acid metabolism in the micropremie. AB - Lipids are structural components of all tissues and are indispensable for cell membrane synthesis. The brain, retina, and other neural tissues are particularly rich in LCPUFAs, affecting neural structural development and function. LCPUFAs serve also as specific precursors for eicosanoid production (prostaglandins, prostacyclins, thromboxanes, and leukotrienes). These autocrine and paracrine mediators are powerful regulators of numerous cell and tissue functions (e.g., thrombocyte aggregation, inflammatory reactions, and leukocyte functions, vasoconstriction and vasodilatation, blood pressure, bronchial constriction, uterine contraction). Dietary lipid intake affects cholesterol metabolism at an early age and is associated with cardiovascular morbidity and mortality in later life. Over recent years, the role of fatty acids in modulating signal transduction and regulating gene expression have been described, emphasizing the complex of fatty acid effects. Dietary fatty acids, especially LCPUFA, can have significant effects in the modulation of developmental processes affecting the clinical outcomes of extremely premature infants. PMID- 10690566 TI - Vitamin metabolism and requirements in the micropremie. AB - Vitamin metabolism and requirements are reviewed for the micropremie (1000 Pounds g birthweight), for parenteral and enteral feedings. Recommendations are presented in table format. Human milk fortifiers and special formulas for the preterm infant are reviewed. For parenteral nutrition, only MVI Pediatric is currently available in the United States. Two millimeters per kilogram is recommended for the micropremie as the most satisfactory method of providing supplemental vitamins in total parenteral nutrition solutions. PMID- 10690567 TI - Trace elements of the micropremie. AB - The current limited understanding of the ontogeny and mechanisms of the metabolism of iron, zinc, copper, selenium, iodine, and manganese in the micropremie are reviewed. The effects of early delivery on these processes are considered, as are the suggested amounts of these trace elements required for micropremies. PMID- 10690568 TI - Water and electrolyte metabolism of the micropremie. AB - There are five problem schemas presented in this article that indicate potential contradictions in therapeutic goals: (1) shock and edema presenting upon premature birth; (2) the hyperosmolar state, problematic in patients less than 750 g birth weight; (3) the respiratory distress syndrome and respiratory failure, often complicated by patent ductus arteriosus; (4) bronchopulmonary dysplasia, resulting from prematurity and mechanical ventilation; and (5) late onset of hyponatremia, sometimes accompanied by growth failure in the recovering premature. These five problems considered together comprise a developmental continuum of illness and recovery, where appropriate fluid management has recently been demonstrated to benefit outcomes greatly. Clinicians over the past decade have encountered all of the fluid and electrolyte nightmares. Although there are many different formulations for treating each of these scenarios, recommending one approach for all patients is likely to be incorrect much of the time. PMID- 10690569 TI - Bone mineral metabolism in the micropremie. AB - Environmental factors, nutritional supplies, hormonal status, diseases, and treatments appear to affect postnatal skeletal growth and mineralization in VLBW infants. Compared with their term counterparts, ELBW infants are at risk of postnatal growth deficiency and osteopenia at the time of hospital discharge. From recent data, DXA is becoming one of the reference techniques to evaluate mineral status, whole-body composition, and effects of dietary manipulations on weight gain composition and mineral accretion in preterm infants. Weight gain and length increases need to be evaluated carefully during the first weeks of life, in the intensive care unit and out of it, in the step down unit. Nutritional survey is required to improve the nutritional supply and to maximize linear growth. As the critical epoch of growth extends, during the first weeks or months after discharge, follow-up and nutritional support need to be provided during the first years to promote early catch-up growth and mineralization. Further studies need to determine precisely the most optimal feeding regimen during this period but also need to evaluate the long-term implications of such a policy on stature, peak bone mass, and general health at adulthood. PMID- 10690570 TI - Bilirubin and jaundice in the micropremie. AB - Although it has been customary to treat neonatal jaundice at lower serum bilirubin levels in low-birth weight infants than in term infants, the threshold bilirubin levels and long-term benefits for early treatment of preterm infants have not been validated. This article summarizes and evaluates existing evidence and strategies for early treatment of bilirubin in micropremies and recommends a conservative but flexible approach to early monitoring and phototherapy for jaundice in extremely low-birth weight infants. PMID- 10690571 TI - Energy expenditure in the extremely low-birth weight infant. AB - Information about energy requirements of extremely low-birth weight infants is sparse, despite the rapidly improving survival rates of this population. Metabolizable energy intake can be estimated from energy balance studies and the percentage of caloric intake that is actually absorbed by these infants is approximately 87%. Data on energy expenditure in extremely premature infants is limited; however, energy expenditure has been shown to increase with postnatal age. Because both intake and expenditure are affected by multiple factors, there is significant variability in estimates of the energy requirements in extremely low-birth weight infants. At present, no valid recommendations can be made regarding optimal energy requirements for the extremely low-birth weight infant, except that their requirements probably exceed those of stable, growing very low birth weight infants, currently estimated at 105 to 135 kcal.kg-1d-1. PMID- 10690572 TI - Intravenous nutrition and postnatal growth of the micropremie. AB - There is a growing body of evidence that early nutritional practices may affect short-term growth and developmental outcome. In addition, they may play a role in determining adult health and disease. There is much that needs to be learned about safe and efficacious nutrient administration in the ELBW population; about techniques to assess the effect of different nutritional strategies; and about the long-term effects of these regimen or development outcome, growth, and disease. PMID- 10690573 TI - Enteral feeding of the micropremie. AB - Clinical practice demands knowledge of gastrointestinal ontogeny and the factors that affect our ability to use enteral feeding in the micropremie. The decisions regarding milk type (when and how it should be given) are considered in the light of current physiologic and clinical evidence. Special considerations apply in the micropremie who is also small for gestational age and NEC must be avoided. Trophic feeding now has an established role, allowing the infant to benefit from enteral feeds even when full nutritive milk feeding is not possible. PMID- 10690574 TI - Human milk feeding of the micropremie. AB - There is increasing evidence that mother's milk is an appropriate feeding even for LBW and VLBW infants. During early neonatal life, supplements in the form of human milk fortifiers or single nutrients may be necessary to maintain an adequate biochemical status especially for sodium, phosphorus, and protein. The ideal amount and balance of supplemental nutrients to add to mother's milk for small premature infants remain unknown. From research to date, it is clear that growth responses to fortified mother's milk fed in early life may not be the most important outcomes in relation to long-term growth and development. Infants who receive human milk in early life may be at reduced risk for developing infections and allergy as well as osteopenia and growth failure. Further studies are needed to provide a better understanding of the role of human milk as the sole source of nutrition in premature infants, especially the micropremie. PMID- 10690575 TI - Personality disorders and depression: comorbidity. AB - Depression and comorbid personality disorders relate to one another in three distinct ways: 1) personality disorders may precede the development of depression and render an individual vulnerable to depression; 2) depression may precede the personality disorder and foster the development of the personality disorder; 3) there may be an interface between personality disorders and depression, which has been deemed depressive personality disorder. This article reviews data on the comorbidity of depression, particularly chronic depression, and personality disorders. This article also reviews data on the effect of comorbid personality disorders on treatment for depression, and the effect that treatment for depression has on personality disorders. Comorbid personality disorders generally do not impede treatment for depression. Successful treatment for depression is associated with improvement in personality disorders. PMID- 10690576 TI - Factors differentiating personality-disordered individuals with and without a history of unipolar mood disorder. AB - The relationship between mood disorders and personality disorders has been of longstanding interest to clinicians. Despite theoretical reasons to do so, virtually no studies have examined factors that discriminate personality disordered subjects with a history of mood disorder (PD/HMD) from personality disordered subjects without a history of mood disorder (PD). This study examined demographic variables, patterns of comorbidity, measures of life functioning, personality traits, and early life experiences differentiating PD/HMD (n = 83) from PD (n = 214). Diagnoses were assigned using structured clinical interviews and a best-estimate procedure. The results suggest that subjects with borderline personality disorder are more likely to have a life history of mood disorder than are subjects with other personality disorders. In addition, PD/HMDs are more likely to receive a diagnosis of anxiety disorder or alcoholism, to have lower Global Assessment of Functioning (GAF) scores, and to have sought treatment than PDs. On self-report measures of personality, PD/HMDs endorse higher levels of trait anxiety and affective lability (e.g., Harm Avoidance, Neuroticism) than do PDs. PD/HMDs are also more likely to report childhood physical and emotional abuse than are PDs, and to describe their parents as using affectionless control. No differences were found between Axis II clusters as a function of mood disorder history. The discussion suggests a potential model in which early environmental stress interacts with constitutional vulnerabilities to put individuals at an increased risk for both mood and anxiety disorders as well as personality disorders. PMID- 10690577 TI - Relationship between depression and borderline personality disorder. AB - The frequent occurrence of depressive symptoms in patients with borderline personality disorder has generated considerable interest in the nature of the relationship between borderline personality disorder and the depressive disorders. Data from the perspectives of phenomenology, biology, family history, course of illness, comorbidity patterns, and treatment response have been brought to bear on the question. Reviews based on research available by 1985 and 1991, respectively, arrived at differing conclusions: (1) that both disorders shared common but non-specific sources, and (2) that the two disorders were unrelated but co-occurred because of the high prevalence of each. Since the time of these reviews, additional evidence has become available from a wider range of biological investigations, better controlled comorbidity studies, studies of the relationship of psychosocial stressors to the course of each disorder and neuroimaging studies. In reviewing the more recent findings, we propose the less parsimonious hypothesis that the disorders co-occur, both because they share some common biological features and because the psychosocial sequella of each can contribute to the development of the other. PMID- 10690578 TI - Defense mechanisms and personality in depression. AB - There is a longstanding belief that personality represents a structure that is stable over time, and changes, if at all, very slowly. Nonetheless, clinical and empirical evidence suggests that in patients with some Axis I disorders, the rate of personality disorders using DSM criteria decreases after treatment, suggesting that personality as assessed by phenomenological systems is state-dependent. An alternative to the DSM phenomenological system of conceptualizing personality is the dynamic concept of character, that is, a predictable pattern of both adaptive and pathological defense mechanisms, and personality organization comprised of object relations, ego strengths, and superego development. Data from this study address the hypothesis that defense mechanisms and personality organization remain relatively stable in patients treated for Axis I disorders, irrespective of clinical improvement. Patients meeting DSM-IV criteria for major depressive disorder (MDD) entered randomized, controlled medication trials. Defensive functioning was evaluated with the Defense Style Questionnaire (DSQ) [Bond et al., 1983: Arch Gen Psychiatry 40:333-338], and personality organization was assessed with the Inventory of Personality Organization (IPO; Clarkin et al., unpublished), both at baseline and at the completion of the clinical trial. Data were analyzed for whether an individual's pattern of defense mechanisms and personality organization were stable over time regardless of response to treatment of MDD. The question was also asked whether a predominant pattern of defense mechanisms or level of personality organization predicts response to treatment or dropout rate. Among treatment responders, nonresponders and drop outs, baseline DSQ scores were similar except for "image-distorting" defenses, which were significantly more prevalent among drop-outs compared to responders (P = .016). Post-treatment DSQ values revealed a significant decrease in "maladaptive" defenses (P = .01) in the entire sample, while intermediate and "adaptive" defenses remained unchanged. This same pattern was found to hold true in treatment responders. When comparing treatment responders and nonresponders at the end of the trial, medication responders used significantly less "maladaptive" defenses than did nonresponders (P = .003), and had a significantly higher, or healthier level of "overall defensive functioning" (P = .04). Baseline and post treatment IPO values did not show significant differences. Results of the study address the question of whether there are personality characteristics that are enduring and that can be appreciated irrespective of an Axis I disorder. PMID- 10690579 TI - Co-occurrence of mood and personality disorders: a report from the Collaborative Longitudinal Personality Disorders Study (CLPS). AB - The purpose of this study was to examine the relationship of subtypes and particular clinical features of mood disorders to co-occurrence with specific personality disorders. Five hundred and seventy-one subjects recruited for the Collaborative Longitudinal Personality Disorders Study (CLPS) were assessed with the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I) and the Diagnostic Interview for DSM-IV Personality Disorders (DIPD-IV). Percent co occurrence rates for current and lifetime mood disorders with personality disorders were calculated. Logistic regression analyses examined the effects of clinical characteristics of depressive disorders (e.g., age at onset, recurrence, symptom severity, double depression, and atypical features) on personality disorder co-occurrence. In comparison with other DSM-IV personality disorders, avoidant, borderline, and dependent personality disorders (PDs) were most specifically associated with mood disorders, particularly depressive disorders. Severity and recurrence of major depressive disorder and comorbid dysthymic disorder predicted co-occurrence with borderline and to a lesser extent research criteria depressive personality disorders. The results are consistent with the view that a mood disorder with an insidious onset and recurrence, chronicity, and progression in severity leads to a personality disorder diagnosis in young adults. PMID- 10690580 TI - Inflammatory bowel disease at the end of its first century. PMID- 10690581 TI - The genetic background of inflammatory bowel disease. AB - Available evidence indicates that genetic factors are essential in providing the susceptibility to the majority of the various forms of inflammatory bowel disease occurring in man. It is also clear that the genetic susceptibility to these diseases is complex, and that more than one gene may predispose (the concept of multilocus/oligogenic inheritance), and likely in different etiologic combinations (the concept of genetic heterogeneity). Paradigms are now available that should lead to the identification of a number of these predisposing genes. These paradigms include the candidate gene approach, systematic genome wide scans, and mouse human synteny. While genome wide scans are currently limited to multiplex family linkage studies, both candidate genes and mouse human synteny can be approached in either linkage or association paradigms. Eventually whole genome association studies will be available as well. Identification of inflammatory bowel disease predisposing genes should lead to their incorporation in studies of natural history, investigation of environmental risk factors, and especially utilization of genetic markers in clinical trials. This will allow us to identify the best therapy available for the individual patient based on their unique genetic constitution. With advances in molecular technology, the search for genes influencing traits and diseases with a complex genetic background, such as the inflammatory bowel diseases, has become a realistic task. Although exogenous or infectious agents may contribute to the pathogenesis or may trigger the onset of disease, and the immune system almost certainly mediates the tissue damage, it is clear from available data that genetic factors determine the susceptibility of a given individual to inflammatory bowel disease (reviewed below). Thus, genetic studies are essential for the delineation of the basic etiologies of the various forms of inflammatory bowel disease and thus can aid in the development of radically new and specific therapies. In this review, we will discuss the importance and complexity of genetic factors in inflammatory bowel disease, methods and problems in the genetic dissection of complex traits, and future directions of genetic studies in inflammatory bowel disease. PMID- 10690582 TI - Inflammatory bowel disease: immunologic concepts. AB - The past decade has been characterized by an enormous progress in the understanding of the pathophysiology of chronic intestinal inflammation. Novel drugs, which include tumor necrosis factor alpha binding proteins (i.e., the monoclonal antibody infliximab), have been introduced into the therapeutic management of Crohn's disease. With our increasing knowledge of disease pathophysiology and the identification of key mediators, it is to be expected that the efficacy of anti-inflammatory therapy will further increase. The exploration of the genetic etiology of inflammatory bowel disease holds great future promises. PMID- 10690583 TI - Etiology and pathophysiology of inflammatory bowel disease--environmental factors. AB - Environmental factors play an important role in the pathophysiology of inflammatory bowel disease. There is a strong and consistent association between smoking and Crohn's disease, and between nonsmoking and ulcerative colitis. Despite extensive research, the exact pathophysiological mechanisms for these associations remain unclear. In spite of this, some clinical trials with nicotine patches showed beneficial effects for the treatment of ulcerative colitis. Associations of Crohn's disease and ulcerative colitis with other environmental factors are weaker like the association with use of oral contraceptives or those less well investigated such as the association with childhood hygiene. Most studies suggesting a potential pathogenetic role of Mycobacterium paratuberculosis or an effect of tuberculostatic therapy in Crohn's disease could not be reproduced by others. Perinatal or childhood infections by viruses like measles are heavily debated, but not proven to be causal for inflammatory bowel disease. Coagulation disorders have been described as protecting from inflammatory bowel disease, suggesting hypercoagulability to be a pathogenetic factor. Some studies described that appendectomy may prevent the onset of ulcerative colitis in man and mice. Other environmental factors such as hydrogen sulfide, tonsillectomy, diet, blood transfusions, and Listeria also require confirmation. There are, however, convincing data from genetic animal models and twin studies that environmental factors as the intestinal bacterial flora interact with susceptible hosts to cause inflammatory bowel disease. Inflammatory bowel diseases have multifactorial etiologies, which require a differentiated approach for treatment and prevention. PMID- 10690584 TI - Diagnostic approach to IBD. AB - Inflammatory bowel disease, with Crohn's disease and ulcerative colitis as the two main disorders, is a heterogeneous group of disorders of unknown etiology. Clinical initial presentation is sometimes misleading and causing diagnostic delay which may be important. Identification of subgroups of patients on the basis of genetic, immunologic and clinical markers will be important for exact diagnosis, but also for new drug trials. The current diagnosis depends on clinical, radiographic, endoscopic and laboratory data. The introduction of serological markers such as pANCA and ASCA will allow an increase in the diagnostic accuracy at initial diagnosis of inflammatory bowel disease and may play a role in defining subgroups of the disease. PMID- 10690585 TI - Complications of inflammatory bowel disease. AB - Complications in inflammatory bowel disease determine the severity of disease as well as the complexities of medical or surgical treatment opportunities. Therefore, in known inflammatory bowel disease, the prevention, the early detection and the adequate therapeutic response to certain complications are important goals in the follow-up of inflammatory bowel disease patients. Disease complications are separated into intestinal and extraintestinal complications. Intestinal complications are somewhat disease specific, which means that they occur exclusively in either Crohn's disease or ulcerative colitis (e.g., enteric fistulas are particularly found in Crohn's disease and toxic megacolon in ulcerative colitis). Most extraintestinal complications occur in both forms of inflammatory bowel disease (e.g., anemia, thromboembolic events or osteoporosis). The current knowledge on pathogenesis, diagnostic tools, prevention and treatment of certain intestinal and extraintestinal complications is reviewed. PMID- 10690586 TI - Chronic inflammatory bowel disease and cancer. AB - Colorectal cancer represents the major cause for excess morbidity and mortality by malignant disease in ulcerative colitis as well as in Crohn's disease. The risk for ulcerative colitis associated colorectal cancer is increased at least 2 fold compared to the normal population and colorectal cancer is observed in 5.5 13.5% of all patients with ulcerative colitis and 0.4-0.8% of patients with Crohn's disease. Established risk factors include long duration of the disease, large extent of the disease, low activity of the disease, young age at onset, presence of complicating primary sclerosing cholangitis or stenotic disease and possibly lack of adequate surveillance, inadequate pharmacological therapy, folate deficiency and non-smoking. Crohn's disease is associated with an increased risk of colorectal carcinoma in patients with long-standing disease, strictures and fistulae under the condition that the colon is involved, tumors of the small intestine may occur occasionally. Extracolonic malignancies are rare, with the exception of biliary tract cancer. Ulcerative colitis associated colorectal cancer typically can occur in the entire colon, is often multifocal and of undifferentiated histology. Stage distribution and prognosis of ulcerative colitis associated colorectal cancer appears to be similar to that of sporadic colorectal cancer with an overall survival of about 40% (15-65%) after 5 years with tumor stage at diagnosis being the most important predictive parameter for survival. Tumor markers helpful for the diagnosis of sporadic colorectal cancer fail to differentiate between inflammatory response and malignant transformation. In contrast the histologic evidence of dysplasia was shown to be a strong indicator of underlying carcinoma or developing malignant transformation. The presence of a surface projection termed dysplasia associated lesion or mass is highly indicative of underlying or associated cancer. While the routinely performed search for dysplasia is hampered by high interobserver variation the demonstration of DNA-aneuploidy or genetic changes which may confirm the ongoing malignant transformation has not yet become clinical routine. The genetic alterations found in ulcerative colitis associated colorectal cancer involve many of the same targets found in sporadic colorectal tumors and include multiple sites of allelic deletion, microsatellite instabilities, and mutations of APC, p53, Ki-ras as well as MSH2 and other genes. The progression of dysplasia to carcinoma is generally accompanied by an accumulation of these mutations and the similarities in the biology of colorectal cancer associated with ulcerative colitis and sporadic colorectal cancer appear to outweigh their difference. In regard to the management of dysplasia and cancer, the role of surveillance programs for the early detection of ulcerative colitis associated colorectal cancer at a curable stage is still under debate. Although these programs failed at tumor prevention and lethal carcinomas are still found inadvertently in patients under surveillance, the majority of surveillance programs could reduce mortality by detecting more cancers at a still curable stage. Current recommendations for surveillance include, therefore, biennial colonoscopy with extensive biopsies after 8-10 years of total colitis or after 15-20 years of left sided colitis. In the presence of cancer or unequivocal high-grade dysplasia and/or dysplasia associated lesion or mass proctocolectomy is considered adequate. The evidence of low-grade dysplasia should be confirmed before proctocolectomy is considered. PMID- 10690587 TI - Immunopharmacology of 5-aminosalicylic acid and of glucocorticoids in the therapy of inflammatory bowel disease. AB - Glucocorticoids as well as 5-aminosalicylic acid have been used successfully in different formulations during the past 40 years for the treatment of both acute and chronic inflammation in inflammatory bowel disease. The mechanism by which the drugs exert their actions are only partially known. Recent studies of the immunoregulation in the lamina propria provide evidence that numerous therapeutic mechanisms contribute to the efficacy of these drugs including the inhibition of arachidonic acid metabolism, a decrease in radical formation by oxygen radical scavenging, an inhibition of both in vivo and in vitro activation of peripheral and intestinal lymphocytes. Moreover direct immunoregulatory effects exerted by the drug may be important in influencing the complex balance of pro-inflammatory mechanisms during active intestinal inflammation. Such effects are the inhibition of both peripheral and intestinal B lymphocyte immunoglobulin secretion as well as the inhibition of pro-inflammatory cytokine production and their binding to receptors. Some of these immunoregulatory effects appear to be mediated by an inhibition of the activation of the nuclear factor kappa B transcription factor family by steroids and (less potent) aminosalicylic acid. Activation of nuclear factor kappa B appears to be pivotal for the sustained upregulation of inflammation molecule expression in many inflammatory diseases. It seems, therefore, most likely that the enormous therapeutic potency of steroids, as well as the anti-inflammatory properties of 5-aminosalicylic acid, are not achieved by a single action of the drug. The complex orchestration of numerous inhibitory interactions with pro-inflammatory principles will add to the therapeutic potential of steroids and of 5-aminosalicylic acid in the treatment of both acute and chronic intestinal inflammation. PMID- 10690588 TI - Treatment of ulcerative colitis. AB - In recent years new standards for the treatment of ulcerative colitis have evolved. This review updates evidence based therapy for the various clinical situations as well as some novel approaches. The literature search was based on Medline, Cochrane database (CD-ROM) and handsearch of relevant papers including quoted literature. There is clear-cut evidence-based support for the use of local 5-aminosalicylates in mild/moderate distal and oral 5-aminosalicylates in extensive ulcerative colitis. The administration of corticosteroids is definitely indicated in severe disease. Fulminant attacks are treated by intravenous cyclosporine or colectomy. In chronic active disease azathioprine is probably helpful. Relapse prevention again is a domain of 5-aminosalicylates or, as a novel development, E. coli Nissle. The various meta-analyses as well as the controlled trials performed in the various clinical situations typical for the manifestations of ulcerative colitis form a solid base of evidence to guide individual treatment decisions. PMID- 10690589 TI - Treatment of Crohn's disease. AB - The treatment of Crohn's disease depends on disease location and disease activity. It can be divided into medical and surgical treatment. While surgery is reserved for complications such as abscesses or failure of pharmacological treatment (fistulae, perianal disease, or strictures) medical treatment aims at induction and maintenance of remission. In order to achieve these goals supportive and therapeutic strategies must be used. Supportive measures include substitution of vitamins, particularly fat-soluble vitamins, and minerals in deficiencies due to resection or disease involvement of the small bowel. All patients on long-term steroids should receive calcium and vitamin D in order to prevent osteoporosis. Therapeutic options include drug treatment (corticosteroids, antibiotics, salicylates, and immunosuppressives), nutrition (parenteral or enteral), and endoscopy (dilatation of strictures). Depending on disease location different pharmacologic preparations of salicylates or corticosteroids should be used, e.g., enemas for distal colitis. The most potent drugs for long-term control are immunosuppressive agents, particularly azathioprine. It is the most widely investigated immunosuppressive agent in Crohn's disease and should be the first line treatment for patients with steroid refractory, chronic steroid dependent, fistulating, and stenosing courses. In the future, more potent drugs and better risk stratification criteria should improve the treatment of Crohn's disease. PMID- 10690590 TI - Future developments in diagnosis and treatment of inflammatory bowel disease. AB - The inflammatory bowel diseases ulcerative colitis and Crohn's disease are probably syndromes rather than single entities. Neither the susceptibility genes nor definite environmental factors have been found thus far. The "immune concept" of these disorders might not include all patients. Consequently immune based new approaches on alternative etiological/pathophysiological pathways may be necessary. New developments in diagnostic techniques will probably improve patient acceptance and may even help to prevent carcinoma development. PMID- 10690591 TI - Trypsin level in gallbladder bile and ductitis and width of the cystic duct. AB - BACKGROUND/AIMS: The change from laparotomy to laparoscopy for cholecystectomy has raised the question of how to manage concomitant bile duct stones. The present-day interest--and controversy--has focused on a transcystic approach reported to be feasible in 66-96% of cases, but without explaining the necessary prerequisite: the widening of the cystic duct. The cystic duct, wide mainly in patients with bile duct stones, has been reported to be highly variable: from strictured to very wide. The present study aims at comparing the trypsin level in the gallbladder bile and the cystic duct morphology and width in patients with and without bile duct stones. METHODOLOGY: A prospective series of 63 gallstone patients, 30 with and 33 without bile duct stones (controls), underwent cholecystectomy and bile duct clearance. The study includes the trypsin level in the gallbladder bile, the width and morphology of the cystic duct, and the size of the gallstones. RESULTS: The patients with bile duct stones had, in contrast to the controls, higher trypsin levels in the gallbladder bile (P < 0.001) and wider cystic ducts (P < 0.001) with more pronounced signs of chronic ductitis. CONCLUSIONS: The obtained results strongly suggest that the increased trypsin level, a sign of reflux of pancreatic juice, caused changes in the cystic duct that facilitate gallstone migration, which also ought to render a transcystic stone extraction feasible. PMID- 10690592 TI - Synchronous carcinoma of the gallbladder in a patient with intrahepatic bile duct carcinoma. AB - An 83-year-old woman, diagnosed as having cholelithiasis, was admitted to the Department of Surgery, Nippon Medical School, with right hypochondrial pain. Ultrasonography and computed tomography revealed a mass in the gallbladder fundus and a hypovascular tumor in the anterior segment of the liver. Magnetic resonance imaging showed stenosis of the intrahepatic bile duct and dilatation of its proximal portion. She was diagnosed as having intrahepatic bile duct carcinoma combined with gallbladder carcinoma. At laparotomy, there was evidence of multiple peritoneal metastases and intraoperative histological examination of the gallbladder tumor revealed adenocarcinoma. Accordingly, only cholecystectomy and needle biopsy of the liver tumor was performed. Histological examination of the gallbladder revealed papillary adenocarcinoma invading the muscularis propria with medullary growth or intermediate stroma. There was no microvessel invasion, no perineural invasion and no lymph node involvement. On the other hand, the liver tumor was a cholangiocarcinoma with a well-differentiated tubular pattern. Therefore, this was a rare case of synchronous carcinoma of the gallbladder associated with intrahepatic bile duct carcinoma. PMID- 10690593 TI - Coexistence of primary biliary cirrhosis and myasthenia gravis: a case study. AB - We present a case that suggests a relationship between primary biliary cirrhosis and myasthenia gravis. A 43-year-old Japanese woman was admitted to the Nagoya City University Medical School, First Department of Internal Medicine with abnormal liver function in August 1991. She had had ptosis of the right eye since 1990. She had not been treated for liver disease. Ptosis of the right eye and hepatomegaly were present. Serum laboratory examinations revealed elevated biliary enzymes and IgM levels; tests were positive for antimitochondrial antibody and antiacetylcholine antibody. Liver histology revealed chronic non suppurative destructive cholangitis and led to a diagnosis of primary biliary cirrhosis. The tensilon test was positive. Electromyography with repetitive motor nerve stimulation revealed a neuromuscular junction defect; i.e., the primary characteristic of myasthenia gravis. The patient was diagnosed with myasthenia gravis. Although the development of myasthenia gravis has previously been reported in patients with primary biliary cirrhosis during D-penicillamine administration, this is a very rare case of the coexistence of both diseases before such treatment. PMID- 10690594 TI - A case of segmental primary sclerosing cholangitis. AB - A 74-year-old man was admitted to the Yokohama City University School of Medicine for investigation of high values of ALP and Y-GTP. Radiographic examinations, including abdominal computed tomography and percutaneous transhepatic cholangiography, strongly suggested bile duct cancer in the hepatic hilus. After left lobectomy, pathological examination disclosed segmental primary sclerosing cholangitis. Clinical examination cannot always distinguish primary sclerosing cholangitis from cancer. We report a case of segmental primary sclerosing cholangitis and discuss the diagnosis and the treatment of this disease. PMID- 10690595 TI - Adenomyoma of the common bile duct: report of a case. AB - We report a case of adenomyoma in the common bile duct accompanied by obstructive jaundice. A 64-year-old woman presented with abdominal pain, fever, appetite loss and jaundice. Endoscopic retrograde cholangiopancreatography revealed possible stenosis in the distal common bile duct. We could not distinguish whether the tumor was benign or malignant based on the clinical presentation, or biochemical, radiographic, or endoscopic investigations. Pancreatoduodenectomy was performed. The histological diagnosis was adenomyoma. The natural history of and optimal treatment for, adenomyoma have not been established. PMID- 10690596 TI - Primary carcinoid tumor of the gallbladder: resection of a case metastasizing to the liver and analysis of outcomes. AB - Gallbladder carcinoid tumor is a rare and aggressive neoplasm, usually lacking specific symptoms, as they typically are unassociated with the carcinoid syndrome, despite frequent hepatic spread. The patient was an 81-year-old man with right upper quadrant pain who underwent radical surgery for carcinoid tumor of the gallbladder with liver metastasis (preoperative diagnosis, carcinoma). We analyzed the outcome of previously reported cases of gallbladder carcinoid. Increasing tumor size and depth of invasion progressively compromised the 2-year survival. These findings underscore the importance of early detection. PMID- 10690597 TI - A new examination for both biliary and gastrointestinal function after pancreatobiliary surgery--single-isotope two-day method. AB - BACKGROUND/AIMS: There are no established tests for both biliary and gastrointestinal function after pancreatobiliary surgery. In this study, mixing of ingested food with bile after long-term postoperative period was evaluated. METHODOLOGY: Sixteen patients having undergone Imanaga pancreatoduodenectomy (postoperative years = 6.2 +/- 2.0) were evaluated concerning physiologic function of biliary tract and residual stomach using gastric emptying and hepatobiliary scintigraphy. RESULTS: At least 24 months after Imanaga pancreatoduodenectomy, postoperative patients had reached 95.8 +/- 4.9% of the pre-illness bodyweight. Gastric emptying half-time (GET1/2) ranged from 9-147 min (mean: 60.6 +/- 35.0). The time of bile excretion from liver to jejunum ranged from 5-45 min (mean: 11.3 +/- 7.8). Asynchrony time ranged from -5-40 min (mean: 11.3 +/- 7.8). There was no significant correlation between GET1/2 and recovery of bodyweight (r = -0.042, P = 0.8809). Similarly, there was no significant correlation between the time of bile excretion and recovery of bodyweight (r = 0.042, P = 0.8791). On the other hand, asynchrony time had a significant inverse correlation with recovery of bodyweight (r = -0.590, P = 0.0146). CONCLUSIONS: Single-isotope two-day method is an original and useful technique to evaluate biliary and gastrointestinal physiologic function after long-term postoperative period. PMID- 10690598 TI - Surgical anatomy of the medial segment (S4) of the liver with special reference to bile ducts and vessels. AB - BACKGROUND/AIMS: Resection of the inferior area of the medial segment (S4a) plus S5 with preservation of the superior area of the medial segment (S4b) is being performed to manage hilar bile duct carcinoma and pT2 type gallbladder carcinoma, and thus, attention has been focused on the surgical anatomy of the medial segment of the liver to identify the specific vessels and bile ducts of the areas of that segment to be resected and to be preserved. METHODOLOGY: Anatomical study of the bile duct, portal vein, middle hepatic vein, and middle hepatic artery to the medial segment branches of the liver (S4) was performed in a total of 171 specimens comprised of 71 adult cadavers, and 100 liver casts. RESULTS: 1) Two main types of bile duct branches of the medial segment (B4) were recognized. Type I included the branches which joined to the left hepatic duct on the hilar duct side (35.5%), and type II included the branches that joined on the peripheral side (54.6%). Several subtypes were also found in both types. The B2-B3 confluence was mostly on the left (41.7%) or posterior (42.7%) to the umbilical portion (UP) of the portal vein, and to the right of the UP (hilar side) in only 15.6%. 2) The portal vein of the medial segment branches (P4): P4a branched from the right angle and upper right border of the UP in every specimen. The most common morphology was 1 large and 2-3 small branches (41%). P4b was almost always found to branch posterior to the UP and lower than P4a, and the most common morphology was 1 large and 0-1 small branches (57.8%). 3) The middle hepatic vein: In 83.2% a common trunk was observed at the confluence with the inferior vena cava, and 8 types of the middle hepatic vein were recognized. 4) The middle hepatic artery: It arose from the left hepatic artery in 61.5%, from of the right hepatic artery in 27.5%, from the proper hepatic artery in 5.5%, and from both the left and the right hepatic artery in 5.5%. CONCLUSIONS: The detailed vascular and bile duct anatomy of S4 is described. This study should be helpful in identifying the specific vessels and bile ducts of the areas of the medial segment to be resected and to be preserved, thereby facilitating resection of the medial segment. PMID- 10690599 TI - Gastric pseudotumor. AB - The authors present a case report of a pseudotumor of the stomach and a brief discussion about this very unusual entity. A 75-year-old female patient was admitted with melena and a large epigastric tumor; she underwent upper gastrointestinal endoscopy, abdominal ultrasound, magnetic resonance imaging, guided needle aspiration and angiography. Preoperative diagnostic hypothesis included a partially thrombosed aneurysm of the splenic artery, pancreatic cystic neoplasm with gastric invasion and pancreatic pseudocyst complicated with hemorrhage. Laparotomy revealed a gastric tumor and the patient was submitted to a radical subtotal Billroth II gastrectomy. Only the pathologic examination revealed the unexpected definitive diagnosis of an organized intramural gastric hematoma. There were no postoperative complications and she remains asymptomatic 10 months after surgery. PMID- 10690600 TI - Factors influencing bowel function after low anterior resection and sigmoid colectomy. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the subjective bowel function after low anterior resection and sigmoid colectomy and to clarify the clinicopathologic factors influencing postoperative bowel habits. METHODOLOGY: Eighty-six patients who underwent low anterior resection and sigmoid colectomy replied to the questionnaire which consisted of 8 categories of bowel symptoms. The patients were divided into 2 groups: good bowel function showing less than half of symptoms (< 4) and poor bowel function showing more than half of symptoms (> or = 4). RESULTS: After low anterior resection, patients were often complicated with incomplete evacuation (75%), bowel movement at night (60%), defecation more than twice a day (46%), and soiling (27%). The mean number of defecation/day and frequency of patients with night stools was significantly higher after low anterior resection than sigmoid colectomy (2.81 vs. 2.18, P < 0.05; and 60% vs. 29%, P < 0.05). Poor bowel function after low anterior resection was frequent in patients with high ligation of the inferior mesenteric artery (82%, P < 0.05), injury to the pelvic autonomic nerve (82%, P < 0.05), and blood transfusion; while poor bowel function after sigmoid colectomy was frequent in patients with resected colon measuring 25 cm or more (81%, P < 0.05). CONCLUSIONS: These results indicate that poor bowel function after low anterior resection is associated with high ligation of the inferior mesenteric artery and injury to the pelvic autonomic nerve; while poor bowel function after sigmoid colectomy correlates with length of the resected colon. Less aggressive surgery is needed to preserve good bowel function. PMID- 10690602 TI - Elevated carcinoembryonic antigen (CEA) levels in a patient with no malignancy. AB - Carcinoembryonic antigen rarely exceeds serum levels of 10-12 ng/mL in benign diseases and has never been found above 24 ng/ml. We report a case in which carcinoembryonic antigen serum levels reached the value of 44.9 ng/ml without any overt reason (after 22 months of follow-up). A decline of the carcinoembryonic antigen to normal ranges was noticed after a radiolabeled anti-carcinoembryonic antigen monoclonal antibody scan was performed. The reason for this phenomenon is unclear. PMID- 10690601 TI - BCL2 and BAX expression in hyperplastic and dysplastic rectal polyps. AB - BACKGROUND/AIMS: Members of the gene family that includes BCL2 and BAX are functionally antagonists in the apoptosis process and they have been observed in normal and neoplastic tissues. The aim of this study is to investigate the combined effects of BCL2 and BAX protein in normal mucosa, dysplastic and hyperplastic polyps of the rectum. METHODOLOGY: We studied BCL2 and BAX protein expression in 40 cases of adenomatous polyps all located in the rectum, with different dysplastic gradings, and the mean time in 10 cases of normal rectal mucosa. RESULTS: BCL2 expression was found more frequently in hyperplastic and in low dysplastic polyps with moderate and strong positivity compared to moderate and severe dysplasia. BAX expression was found in normal mucosa in hyperplastic and dysplastic polyps, the immunoreactivity was prevalently moderate and strong. CONCLUSIONS: These preliminary data suggest that BCL2 and BAX confirm a probably different role in apoptosis. Nevertheless, it is important to know the relation between the molecular pathways of apoptosis, the defective mismatch repair and the tumor suppressor genes associated with an increased mutation rate in cancerogenesis of the colorectum. PMID- 10690603 TI - Occult colon cancer in a patient with an unexplained episode of pulmonary embolism. AB - The association between venous thromboembolism and cancer has been widely documented and the main factor responsible for cancer-induced venous thromboembolism is considered mostly linked to a hypercoagulation state induced by the cancer itself. There is no consensus on investigative strategies for occult cancer in a patient with a thrombophilic condition. We report a patient who manifested an isolated episode of pulmonary embolism without specific evident sources of venous thromboembolism. The routine clinical and laboratory work-up to detect an occult cancer did not reveal any malignancy. A history of duodenal ulcer in association with a recent slight alteration in bowel habits led us to perform an esophagogastroduodenoscopy which was negative for malignancy, and a barium enema followed by colonoscopy, which revealed the presence of a tumor limited to the large intestine. An unexplained clinically evident hypercoagulation state, even in the presence of mild clinical symptoms, needs more thorough diagnostic strategies when simple methods of screening for occult cancer are negative. PMID- 10690604 TI - A prospective randomized trial from Turkey comparing octreotide versus injection sclerotherapy in acute variceal bleeding. AB - BACKGROUNDS/AIMS: Bleeding from gastroesophageal varices continues to be a life threatening complication of chronic liver diseases and portal hypertension. The purpose of this randomized prospective study is to compare the efficacy of octreotide administration and emergency injection sclerotherapy for the control of actively bleeding esophageal varices and prevention of early rebleeding in patients with cirrhosis. METHODOLOGY: A total of 66 episodes of endoscopically proven active variceal bleeding in 52 patients were included in this study. Following admission to the hospital, the patients were resuscitated with blood and plasma, and fiberoptic endoscopy was performed within 2 hours. Thirty-six bleeds in 28 patients and 30 bleeds in 24 patients were randomized to endoscopic variceal sclerotherapy (1% polidocanol) and to octreotide infusion (at 50 micrograms/h for 12 hours following the initial 50 micrograms i.v. bolus), respectively. RESULTS: Bleeding was initially controlled within 6 hours in 75% of episodes by endoscopic variceal sclerotherapy and in 73.3 by octreotide infusion (P > 0.05). There were no significant differences between the 2 groups in early rebleeding (within 72 hours of randomization) (22% vs. 22.7%), blood transfusion (4.2 +/- 1.8 units vs. 4.8 +/- 2.9 units), or hospital mortality (3.6% vs. 3.3%). Treatment failed in 9 episodes (25%) in the sclerotherapy group and in 8 episodes (26.7%) in the octreotide group. CONCLUSIONS: We consider that Octreotide would appear to be as effective as sclerotherapy in both the early control of variceal hemorrhage and in the prevention of early recurrent bleeding and should therefore be considered the treatment of choice in those centers where 24-hour endoscopy is not available. Furthermore, even in hospitals that do have a 24-hour endoscopy service there is good evidence that octreotide therapy should be commenced as soon as a patient enters hospital with a suspected variceal bleed to achieve rapid homeostasis. When initial hemostasis is achieved, elective endoscopic therapies can be undertaken with greater success. PMID- 10690605 TI - Comparison between digestive endoscopy and 24-hour esophageal pH monitoring for the diagnosis of gastroesophageal reflux esophagitis: "presentation of 100 cases". AB - BACKGROUND/AIMS: We present the results obtained from 100 new cases of clinical esophagitis caused by gastroesophageal reflux at the Hospital of Caldas and at the Service of Gastroenterology of VIME (Endoscopical Video Medicine) in Manizales, Caldas, Colombia; between the months of June and November of 1996, evaluated by digestive endoscopy and classified based on the New Savary-Miller 5 Grade Classification. METHODOLOGY: The patients were selected based on the presence of symptomatology suggestive of esophagitis caused by gastroesophageal reflux; an endoscopy was performed followed by 24-hour esophageal pH monitoring. The patients were grouped according to their grade of esophagitis in the New Savary-Miller Classification. The central analysis was focused on determining the existing relationship between the observed esophagitis and the results obtained by the 24-hour esophageal pH monitoring. RESULTS: Findings show that 51% and 48% of patients with esophagitis grades 1 and 2 had a normal DeMeester's score (< 14.7) in channel 1. In channel 2 we found normal scores in 86% and 82% of esophagitis grades 1 and 2, respectively. CONCLUSIONS: We ask whether the average level of pathological reflux of 14.7 can be extrapolated to our population; also whether endoscopical overdiagnosis of esophagitis caused by gastroesophageal reflux exists, or if non-recognized causes of esophagitis exist. Another question is if it is justified to order 24-hour esophageal pH monitoring in patients with grades 1 and 2 esophagitis. PMID- 10690606 TI - Diverticular disease of the small bowel. AB - BACKGROUND/AIMS: The clinical picture of small bowel diverticula has not been well-clarified because of its infrequent incidence and limited case number in most published reports. We evaluated a large series of small bowel diverticula and further compared the clinical picture of the diverticula at different parts of small bowel. METHODOLOGY: The medical records of 88 patients with diverticular disease of the small bowel were reviewed from 1979-1997. This study included all diverticula from duodenum to ileum. RESULTS: The most common small bowel diverticulum was duodenal diverticulum (45%), followed by Meckel's diverticulum (23%). The most common clinical presentation was abdominal pain (48.9%), followed by gastrointestinal bleeding (29.5%). However, among the Meckel's diverticula, gastrointestinal bleeding (60%) was the most common presentation. The accurate diagnostic rate for diverticula, overall, was 25.0%. Thirty-nine (44.3%) of the diverticula were found incidentally. Twenty-three cases (26.1%) were diagnosed by gastrointestinal barium study. Thirty-eight (42.1%) diverticula were left alone without any sequela, and the remaining 55 (56.8%) diverticula needed either bowel resection (30.7%) or diverticulectomy (26.1%). All of the Meckel's diverticula were treated by surgery. Postoperative complication occurred in 9 (10.3%) patients. Hospital mortality rate was 3.4%. CONCLUSIONS: Duodenal diverticulum was the most common small bowel diverticulum. Abdominal pain and gastrointestinal bleeding were the most common clinical presentations. The small bowel diverticula, except for Meckel's diverticulum, did not need to be treated if there were no significant symptoms. PMID- 10690607 TI - Changing pattern of intestinal obstruction in Accra, Ghana. AB - BACKGROUND/AIMS: Over the past 50 years acute intestinal obstruction has remained among the commonest causes of the acute abdomen, along with peritonitis, appendicitis and gastrointestinal perforations. However several observers have noticed over the past 2 decades a shift in the etiological spectrum. The study aims at ascertaining the precise nature of the change. METHODOLOGY: Published data on intestinal obstruction from the Korle Bu Teaching Hospital over the past 50 years were reviewed, as the consistent report format permitted ready comparison of the series. Emphasis was placed on clinical features, established cause and resulting complications. RESULTS: The case load of intestinal obstruction has dwindled over the past 3 decades, accounting for 0.7% of all hospital admissions compared with 1.4% 30 years earlier. External hernias together with adhesive bands still constitute the bulk of presenting cases, but the proportions have changed with strangulated hernias accounting for 59.8% instead of 77.6%. The change has paralleled a rise in elective hernia day case surgery. The incidence of intussusception has almost doubled (7.4% from 4.0%) and it still afflicts the very young. Colonic neoplasms have been commoner over the past 2 decades although the incidence (3.3%) falls short of Western figures. Overall mortality has remained unchanged at 9.4% and this has been associated with a rise in resection rates to 18.3%. CONCLUSIONS: The patterns of intestinal obstruction have been much influenced by changing attitudes regarding elective hernia surgery and evolving financial policies. PMID- 10690608 TI - Reduced levels of coagulation factor XIII in patients with advanced tumor disease. AB - BACKGROUND/AIMS: Coagulation factor XIII, which induces the stabilization of fibrin the final step in the coagulation cascade, has various physiological effects. Among these, its beneficial effect in gastrointestinal bleeding episodes is well known. With the exception of inflammatory bowel disease, however, few data are available about this effect, particularly with regard to its role in diffuse bleeding in tumor patients. The study was designed to carry out prospective follow-up investigations, gathering data concerning factor XIII levels in patients with advanced gastrointestinal tumors and evaluating the course of the disease as well as the incidence of bleeding. METHODOLOGY: Sixty patients (22 women, 38 men; median age: 60; range: 29-79) with advanced gastrointestinal tumors were followed-up prospectively. Factor XIII levels were measured using chromogenic substrate. The correlation between the FXIII level and the patients' survival was analyzed using the Cox model. RESULTS: Factor XIII deficiency (below 70%) was seen in only 7 patients (11.6%), 6 of whom died within a median of 1.5 months after the measurement. In all patients however, there was a significant correlation (P = 0.0133) between FXIII levels and the risk of death. Four bleeding episodes occurred in 3 patients, three times with FXIII levels being below the lower normal range. When substitution was attempted, it was only successful in 1 patient in whom the FXIII level was reduced. CONCLUSIONS: FXIII may have predictive value as a marker for the prognosis in these patients with advanced tumor disease. Bleeding episodes were rarely seen, but when they do occur they may be associated with reduced levels of FXIII, and substitution may be beneficial as an adjunct or even as the sole therapeutic intervention. PMID- 10690609 TI - Management of patients with HCV infection poorly tolerant to recombinant interferon alpha. AB - BACKGROUND/AIMS: To evaluate leukocyte interferon-alpha tolerability and efficacy in the retreatment of patients poorly tolerant to recombinant interferon-alpha. METHODOLOGY: Patients with chronic hepatitis C, poorly tolerant to a previous interferon-alpha treatment (118 patients; 73 "relapsers": Group I; 45 "non responders": Group II) were retreated with 6 MU tiw of leukocyte interferon-alpha for 6 months and then followed-up for 12-34 months. Only patients with complete regression of any previous interferon-related adverse event were included. RESULTS: Three patients dropped out due to recurrence of a severe depressive syndrome. In 86/115 patients (75%) no significant lifestyle changes versus baseline were observed during retreatment, while 29 subjects experienced a moderately negative interference on their living habits. The different influence on the patients' quality of life of leukocyte interferon in comparison with the previous treatment was significant (P < 0.001). In 98 patients the interferon related adverse events significantly decreased. After 12 months of follow-up, a sustained biochemical response was observed in 40 patients (Group I:31; Group II:9), and a persistent virological response in 28 (Group I:23; Group II:5). CONCLUSIONS: The good compliance with leukocyte interferon administration shown by poorly tolerant patients, non-responders/relapsers to recombinant interferon, permitted a retreatment with full doses, so increasing the chance to obtain a larger number of sustained responses. PMID- 10690610 TI - Surgical outcome in cirrhotic patients with hepatitis C-related hepatocellular carcinoma. AB - BACKGROUND/AIMS: The aim of this study was to clarify the surgical outcome in cirrhotic patients with hepatitis C-related hepatocellular carcinoma (HCC). METHODOLOGY: The surgical outcome of 26 HCVAb-positive cirrhotic patients with hepatitis C antibody (the C-related HCC group) and 18 HCVAb-negative cirrhotic patients with (the non-C-related HCC group) undergoing hepatectomy for HCC were compared. The C-related HCC group was HCVAb[+], HBsAg[-] for hepatitis B surface antigen in 25 patients and HCVAb[+], HBsAg[+] in 1, and the non C-related HCC group was HCVAb[-], HBsAg[+] in 15 and HCVAb[-], HBsAg[-] in 3. RESULTS: Preoperative aspartate and alanine aminotransferase in the C-related HCC group were significantly (P < 0.01) higher than in the non-C-related HCC group. There were no significant differences in the operative method, intraoperative blood loss and weight of resected liver or pathological data between the 2 groups. In the recurrence pattern, the incidence of multicentric occurrence in the C-related HCC group (53.3%) was significantly (P < 0.05) higher than in the non-C-related HCC group (7.7%). The mortality rate in both groups was 0% and no operative death was encountered. The crude survival and disease-free survival rates at 3 years were similar: 80.8% and 57.7% in the C-related HCC group and 77.8% and 55.6% in the non-C-related HCC group, respectively. CONCLUSIONS: Although surgically treated cirrhotic patients with C-related HCC showed worse preoperative hepatitis status and a higher incidence of recurrence due to multicentricity compared with non-C-related HCC, the mortality and prognosis of patients with C-related HCC did not differ from that of non-C-related HCC. The indication of hepatic resection and consideration for the high incidence of postoperative multicentric occurrence in the patients with C-related HCC should therefore be more careful than in patients with non-C-related HCC. PMID- 10690611 TI - Management and outcome of liver recipients with post-transplant lymphoproliferative disease. AB - BACKGROUND/AIMS: The possibility of development of post-transplant lymphoproliferative disease by patients receiving immunosuppressive therapy is well known. However, elective treatment and outcome remain controversial. We reviewed the management and outcome of our patients with post-transplant lymphoproliferative disease. METHODOLOGY: Records of 457 patients who underwent orthotopic liver transplantation from 1986 to 1997 were analyzed. Patients who developed post-transplant lymphoproliferative disease were reviewed retrospectively. Incidence, clinical presentation, risk factors and outcomes were examined with special emphasis on ductopenic rejection and hilum involvement. RESULTS: Eleven patients developed a post-transplant lymphoproliferative disease (2.4%). These were B-cell non-Hodgkins lymphoma, Epstein-Barr virus-associated in all cases. Five patients (45.5%) received monoclonal antibodies or antithymocyte globulin. Seven patients (63.6%) developed a lymphoproliferative disease before 9 months post-transplant and 4 recipients (36.4%) after 20 months. No late lymphomas regressed after withdrawal from immunosuppression. Six patients (54.5%) were treated with chemotherapy. Eight patients (72.7%) had a tumoral remission. Five patients (45.5%) developed chronic rejection after immunosuppressant discontinuation. Four of them died as a consequence of ductopenic rejection and retransplantation was required in another; 2 died due to graft hilum infiltration. Five patients (45.5%) are alive after a follow-up of 36.5 +/- 32 months (range: 4-77 months). CONCLUSIONS: Patients with post-transplant lymphoproliferative disease require a close follow-up in order to promptly treat conditions that could lead to death. In our series, these were more closely associated with a failing transplanted organ than with the lymphoma itself. PMID- 10690612 TI - Role of iron load on fibrogenesis in chronic hepatitis C. AB - BACKGROUND/AIMS: In chronic viral hepatitis, an enhanced iron load is related to lower response to interferon. Furthermore, iron, through the production of oxygen radicals, may stimulate hepatocyte necrosis and the activation of cells responsible for synthesis and deposition of extracellular matrix. We investigated the relationship between iron load, evaluated by serum assays, and liver fibrogenesis in chronic active viral hepatitis. METHODOLOGY: Serum iron, ferritin, transferrin saturation and serum markers of hepatic fibrogenesis (Laminin and the amino-terminal peptide of procollagen III-NPIIIP-) were assayed in 102 patients (47 females, 55 males, mean age 42.48 years) affected by chronic hepatitis C virus and in 81 healthy controls (47 males, 34 females). In hepatitis C virus patients (studied before alpha-interferon treatment) a semiquantitative score for portal inflammation, necrosis and fibrosis was applied to liver biopsy. RESULTS: Serum indices of iron load were higher in hepatitis C virus patients than in controls, and were higher in cirrhotic than in chronic hepatitis cases. Ferritin and serum iron were positively correlated with NPIIIP and laminin; moreover cases with ferritin levels over the normal limit for sex and age had higher levels of NPIIIP and laminin than cases with normal or poor iron status. CONCLUSIONS: Our data suggest that even a mild increase of iron load stimulates hepatic fibrogenesis, probably adding oxygen free radical injury to the damage of viral infection. PMID- 10690613 TI - Experimental and clinical studies on liver regeneration following transcatheter portal embolization. AB - BACKGROUND/AIMS: We studied compensatory hypertrophy following transcatheter portal embolization experimentally in dogs and clinically under the condition of cholestasis. METHODOLOGY: Experimental study: Sixteen dogs were used for this study. Transcatheter portal embolization was performed in the left lobes (70% of the total liver) using Gelfoam powder in dogs with 2-week obstructive jaundice. Liver weight, liver blood flow and the intracellular adenosine triphosphate content of isolated hepatocytes were measured after transcatheter portal embolization. Clinical Study: transcatheter portal embolization of the right portal branch was performed in 13 patients with cancer of the biliary tract and 3 patients with hepatocellular carcinoma before (extended) right lobectomy, using Gelfoam powder and thrombin. Six patients who had a total bilirubin level > 5 mg/dLunderwent a percutaneous transhepatic biliary drainage before transcatheter portal embolization. Liver function tests, a volumetric study with computed tomography and immunohistochemical staining for profilerating cell nuclear antigen and apoptosis in the resected livers were performed. RESULTS: Experimental study: The weight ratio of the non-embolized lobes to the total liver, 2 weeks after transcatheter portal embolization in the dogs with jaundice, was significantly lower than that of the normal dogs with transcatheter portal embolization (40.5 +/- 4.5% vs. 47.6 +/- 3.2%), but significantly larger than that of the dogs without transcatheter portal embolization. The cellular adenosine triphosphate content and tissue blood flow in the embolized lobes were significantly lower than those in the nonembolized lobes in the normal and cholestatic livers. Clinical study: The postoperative course in all patients was uneventful, with no serious complication or liver dysfunction. Extended right lobectomy with caudate lobectomy was equivalent to 65% before transcatheter portal embolization and to 56% after, transcatheter portal embolization owing to compensatory hypertrophy of the left lobe. However, there was no significant difference in liver volume in the patients with or without obstructive jaundice. Apoptosis was observed in the embolized lobe. CONCLUSIONS: Preoperative transcatheter portal embolization with percutaneous transhepatic biliary drainage for the purpose of liver regeneration would be useful for treating extended hepatectomy with obstructive jaundice. PMID- 10690614 TI - 13CO2 excretion in breath of normal subjects and cirrhotic patients after 13C aminopyrine oral load. Comparison with MEGX test in functional differentiation between chronic hepatitis and liver cirrhosis. AB - BACKGROUND/AIMS: Liver function can be evaluated using 13C breath tests that explore liver Cytochrome P450 activity. Aminopyrine is one of the first compounds used in liver function testing. Lidocaine metabolism to monoethylglycinexylidide is also a valid tool to assess liver function. Although liver Cytochrome P450 metabolizes both compounds, lidocaine metabolism is flow-dependent while aminopyrine metabolism does not depend on liver blood flow. METHODOLOGY: The 1st part of the study evaluated the appearance and disappearance rate of 13CO2 in the breath of both normal subjects and in cirrhotic patients, so as to establish optimal sampling times and to evaluate the amount of time needed before performing a subsequent breath test. The 2nd part of the study compared the aminopyrine breath test with the monoethylglycinexylidide test in patients with chronic hepatitis or cirrhosis. RESULTS: Complete 13CO2 disappearance was recorded 24 hours after the test in normal subjects, while it took 3 days to disappear from the breath of cirrhotic patients. Breath sampling at 60, 120 and 180 min were equally valid in differentiating chronic hepatitis from cirrhosis. The aminopyrine breath test and monoethylglycinexylidide test showed a good yet not close correlation. CONCLUSIONS: This study showed that in cirrhotic patients a 13C breath test can be performed 3 days after the previous one. In chronic hepatitis and cirrhotic patients, the aminopyrine breath test and the monoethylglycinexylidide test evaluated similar, but not identical, hepatic subfunctions, suggesting that multiple 13C breath test using different substrates could explore liver function better. PMID- 10690615 TI - Prognostic factors following liver resection for hepatic metastases from colorectal cancer. AB - BACKGROUND/AIMS: We aimed to identify prognostic factors that may allow better patient selection for liver resection for colorectal liver metastases. METHODOLOGY: A retrospective analysis of the files of 120 patients undergoing liver resection for colorectal metastases between 9/85 and 12/96 was performed. Survival and disease-free survival were calculated, and a uni- and multivariate analysis for the prognostic impact of various perioperative factors on survival was performed. RESULTS: Perioperative morbidity and mortality were 28.3% and 5.8% respectively. Median overall survival was 30 months with a 5-year survival rate of 31%. Radicality was the prime prognostic determinant. In patients with R0 resection, a liver metastasis of > 3.5 cm in diameter was the only independent factor associated with an adverse prognosis. CONCLUSIONS: Liver resection for colorectal liver metastases should be attempted if complete resection with clear margins is feasible and may be especially beneficial in patients with small (< or = 3.5 cm) lesions. PMID- 10690616 TI - The effect of albendazole on the prevention of secondary hydatidosis. AB - BACKGROUND/AIMS: Secondary hydatidosis and recurrence are serious complications in hydatid surgery. Although medical treatment and current surgical techniques are more effective in the prevention of cyst formation resulting from spillage of cystic liquid, secondary hydatidosis is still surgically important. Albendazole, a derivative of benzoimidazole, is the most commonly used drug in the medical treatment of echinococcosis. The effectiveness of pre-operative prolonged or single dose applications is supported by the literature. METHODOLOGY: Twenty-two cases of hepatic hydatidosis are evaluated and treated by surgery. Perioperative albendazole treatment was given in a dose of 12-15 mg/kg/day in 4 divided doses. The treatment began 5-20 days before the surgery and continued 3-7 months in a cyclic monthly form, until latex agglutination tests were negative. In the postoperative period, hematological, ultrasonography and computed tomography scan evaluation was carried out. The follow-up period for 21 patients was 6-31 months (mean: 20.52 months). RESULTS: There was no secondary hydatidosis, recurrence or mortality in this study. Early and late morbidity rates were 4.54% and 13.63% respectively. CONCLUSIONS: Our results support that perioperative albendazole treatment is effective in the prevention of secondary hydatidosis. PMID- 10690617 TI - Immunological analysis in xenogeneic fetal porcine liver transplantation into a beagle. AB - BACKGROUND/AIMS: While allogeneic organ transplantation has been performed safely, a major barrier in xenogeneic transplantation is how to inhibit hyperacute rejection. METHODOLOGY: We challenged xenogeneic fetal liver transplantation from pig to dog. The graft was investigated by immunohistochemical analysis on recipient's IgG, IgM and C3. RESULTS: In 1 of 4 cases, the graft escaped hyperacute rejection for about 4 hours after transplantation, however, the recipient died next day due to hemorrhage from the torn capsule of the liver due to the arterial blood pressure of the recipient. Histologically, the parenchyma showed good countenance and no congestion nor hemorrhage was shown in the vessels. On immunohistochemical analysis, canine IgG, IgM and C3 were deposited on the sinusoidal epithelium of the fetal liver more moderately than that of adult control. Fetal porcine liver showed less expression of major histocompatability complex class I antigen than that of the adult one. CONCLUSIONS: We consider that the hyperacute rejection occurred more slowly in xenogeneic fetal liver transplantation than in the adult one due to not only less expression of major histocompatability complex class I, but also lower expression of the epitope recognized by a natural antibody of the recipient. PMID- 10690618 TI - Marginal donors in liver transplantation. AB - The use of marginal donors has become very common in many liver transplantation units due to the increase in the number of possible recipients. Experience has shown that previous donor protocols were too restrictive. Meanwhile, formerly unknown factors influence current donor evaluation. Different donor factors affecting the outcome of transplantation have been studied. Current absolute contraindications are severe macrosteatosis, long ischemia, sepsis, some viral infections and extra-CNS malignancy. Old age, mild to moderate steatosis, long ICU stay, altered liver function tests, hypernatremia, hypotension and pressors, moderately prolonged ischemia and sex mismatch are usually considered relative contraindications. The result of this wider acceptance policy has been an increasing number of usable livers without deleterious influences on graft and patients survival. PMID- 10690619 TI - Fat absorption after pylorus-preserving pancreatoduodenectomy reconstructed with Billroth II pancreaticojejunostomy or Billroth I pancreaticogastrostomy. AB - BACKGROUND/AIMS: The aim of this study was to determine whether Billroth I pancreaticogastrostomy (PG-I) or Billroth II pancreaticojejunostomy (PJ-II) after pylorus-preserving pancreatoduodenectomy is associated with better postoperative fat absorption, based on residual pancreatic exocrine function. Several reconstructive operations have been employed after pylorus-preserving pancreatoduodenectomy to maximize postoperative nutrition. However, no single institution study has been published comparing the reconstructive procedures with respect to digestion and absorption of fat. METHODOLOGY: Fat absorption was studied using the 13C-trioctanoin breath test in patients who were grouped according to the degree of fibrosis of the pancreatic remnant, which was determined by histologic examination of the resection specimen. The fibrosis was graded: grade 0, < 10% fibrosis; grade 1, 10-30% fibrosis; and grade 2, > 30% fibrosis. There were 22 patients in the PG-I group and 22 patients in the PJ-II group. RESULTS: There were no significant differences between the PG-I and PJ-II groups in the cumulative excretion of labeled carbon dioxide in the patients with grade 0 pancreatic fibrosis. The cumulative excretion in the PG-I group was better than in the PJ-II group in the patients with grade 1 and grade 2 pancreatic fibrosis. CONCLUSIONS: Fat absorption after PG-I is superior to that after PJ-II in patients with disordered exocrine function of the pancreatic remnant. Billroth I pancreaticogastrostomy allows more effective utilization of the exocrine enzymes of the pancreatic remnant due to elimination of the blind loop characteristic of the Billroth II pancreaticojejunostomy. PMID- 10690620 TI - Appraisal of two-staged pancreatoduodenectomy: its technical aspects and outcome. AB - BACKGROUND/AIMS: Leakage from the pancreaticoenteric anastomosis after pancreatoduodenectomy is closely associated with intraabdominal hemorrhage, thus contributing to mortality. Recently, two-staged pancreatoduodenectomy including exteriorization of the pancreatic juice and second-look pancreaticojejunostomy was performed in high-risk patients. METHODOLOGY: The authors reviewed 24 patients who underwent two-staged pancreatoduodenectomy from November 1994 to April 1999. RESULTS: Oral intake could be instituted on the 6th (mean) postoperative day. In 23 of the 24 patients, the pancreatic juice leakage stopped within a mean of 10 days without any complications. In the remaining 1, the leakage lasted over 4 weeks and intraabdominal bleeding from the gastroduodenal artery occurred. The median interval between pancreatoduodenectomy and the second operation was 124 days (range: 93-323 days). In 15 patients, a stent tube was placed at the site of pancreaticojejunostomy: 1 patient developed acute pancreatitis due to dislocation of the stent tube, in 3, pancreatic juice leakage necessitated exteriorization of the juice, and the remaining 11 recovered uneventfully. In the other 9 patients, the pancreatic juice was exteriorized: 1 patient had leakage and the other 8 recovered uneventfully. Overall, there was no mortality. CONCLUSIONS: Our two-staged pancreatoduodenectomy is considered to make pancreatoduodenectomy performable safely without any mortality. This procedure is recommended for selected patients, including those who require concomitant major hepatectomy or resection of other organs or who have liver cirrhosis, and may be indicated for patients who have a soft and fragile pancreas or pancreatic trauma. PMID- 10690621 TI - Intraductal papillary-mucinous tumors: an entity which is infrequent and difficult to diagnose. AB - BACKGROUND/AIMS: Intraductal papillary-mucinous tumor of the pancreas is currently considered to be a tumor which is an entity of its own, different from classic pancreatic ductal carcinoma. It is basically characterized by slow growth and low malignancy potential, as well as by the production of mucin. The aim of this study is to contribute to world literature some clarification of its natural history, clinical presentation, the most useful diagnostic tests, methods of detection of stromal invasion and handling of treatment. METHODOLOGY: Of 297 pancreatectomies undertaken at the "12 de Octubre" hospital between May 1985 and January 1998, only 1 case of Intraductal papillary-mucinous tumor was found. We have revised 127 cases published in 10 series over the last 10 years. We also contribute a review of our own case. RESULTS: These tumors, which are very infrequent, produce non-specific symptoms, with long latency periods from the first symptom up to stromal invasion. Endoscopic retrograde cholangiopancreatography showed alterations in 100% of cases where this was undertaken. Tumor-related mortality was zero amongst patients with non-invasive tumor who underwent surgery. None of the cases published presented upper gastrointestinal hemorrhage. This indicated the correct surgery and led us to our diagnosis. CONCLUSIONS: We confirm the low frequency and difficulty of diagnosis, the sensitivity of endoscopic retrograde cholangiopancreatography, the difficulty of early detection of stromal invasion, and the high survival rate in cases where resection is done before this occurs. Early diagnosis and treatment is therefore of utmost importance. PMID- 10690622 TI - Introduction of proton pump inhibitors--consequences for surgical treatment of peptic ulcer. AB - BACKGROUND/AIMS: This retrospective study analyzes the influence of different factors on morbidity and mortality after surgical treatment of peptic ulcer. METHODOLOGY: At the Municipal Hospital of Offenbach, Germany, from 1985-1996, 485 patients underwent surgery. RESULTS: Of the 485 patients, 70.7% (343) were diagnosed to have duodenal ulcer and 29.2% (142) had suffered from gastric ulcer. During this period, 79.2% (384) of the operations were performed under emergency conditions because of acute complications (56% of these with perforation, 20% with penetration, 24% with ulcer bleeding), whereas the rest was done electively. Two hundred and ninety-one (60%) patients were male, the average age was 59 years and 71.7% (348) of the patients had certain concomitant diseases. We observed complications in 48% of the cases with a total postoperative mortality of 21%. CONCLUSIONS: Between 1985 and 1996 the total number of ulcer surgeries performed at the Municipal Hospital Offenbach per year has stayed almost constant. However, a definite increase of acute operations in addition to a decrease of elective interventions was noticed. The dissatisfying results of surgical treatment of peptic ulcer after the introduction of proton pump inhibitors seems to be the consequence of the negative selection of patients mentioned above. A connection could be proved between the age and condition of the patient, the type of the surgical intervention (acute or elective) and the morbidity and mortality after the surgery. PMID- 10690623 TI - Neutrophil functions and cytokine production in patients with gastric cancer. AB - BACKGROUND/AIMS: One of the most important factors in the prevention of postoperative infection is the patient's own capacity to protect against infection. Neutrophils play a major role in this protection through phagocytosis and superoxide generation. Inflammatory cytokines are suitable for estimating the degree of surgical stress. The present study was designed to elucidate whether neutrophil functions are impaired in gastric cancer patients, and are related with cytokine production after surgery. METHODOLOGY: Phagocytosis and superoxide generation by neutrophils was studied in 84 patients with gastric cancer by flow cytometry. IL-6, IL-8 and tumor necrosis factor alpha were studied in 18 patients with gastric cancer by enzyme-linked immunosolubent assay. RESULTS: In gastric cancer patients phagocytosis was not impaired, whereas superoxide generation was lower than benign diseases and it was inhibited relative to the clinical stage. Moreover, superoxide generation was correlated with the nutritional parameters and was more suppressed in 7 patients who suffered from postoperative infection than in 40 patients whose postoperative course were uneventful. The fluctuation of superoxide generation correlated well with the serum cytokine levels in the postoperative course and its correlation was clarified in vitro. Nine patients with gastric cancer received intravenous hyperalimentation, and their superoxide generation was increased. CONCLUSIONS: Superoxide generation by neutrophils was suppressed in gastric cancer patients and it is suggested that nutritional support prevents postoperative infection via the augmentation of superoxide generation. PMID- 10690624 TI - Angiogenesis inhibitor, TNP-470, suppresses growth of peritoneal disseminating foci. AB - BACKGROUND/AIMS: Angiogenesis is critical not only for growth of primary tumors but also for cells established at distant organs. We investigated the effects of angiogenesis inhibitor, TNP-470, on the establishment and growth of intraperitoneally inoculated human gastric cancer cell line, MKN-45, and survival of nude mice with this tumor. METHODOLOGY: Human gastric cancer cell line, MKN 45, were injected into the peritoneal cavity of an ICR nude mouse and a model of peritoneal dissemination was developed. TNP-470 was injected subcutaneously every other day from day 1 until sacrifice or death. The effects of TNP-470 on MKN-45 cells were also examined in vitro. RESULTS: Although the number of disseminated foci was not significantly different, the maximum size was significantly smaller in a TNP-treated group than those of a control. Survival time was significantly longer in a TNP-treated group. TNP-470 demonstrated no growth inhibition of MKN45 cells in vitro. CONCLUSIONS: Those results suggested that anti-angiogenic agent, TNP-470, might be effective in treating peritoneal dissemination of gastric cancer by inhibiting growth of the seeded tumor cells on the peritoneum. PMID- 10690625 TI - CAg 25: a novel tumor-associated mucin antigen. PMID- 10690626 TI - Measles IgA in the nasal washings of adult volunteers and children immunized intranasally with measles vaccine L-16. AB - Earlier it was determined that inspiration of aerosolized measles vaccines may be as effective as its injection in induction of measles antibodies formation. In the beginning of measles infection the measles virus penetrates through mucosa of nose, mouth or eyes of children. So it seems rational to use a nasal spray of measles vaccine to induce "mucosal immunity" in the nasopharinx. Local IgA measles antibodies formation was observed only after measles vaccine spray immunization of 6-7 year old children or adult volunteers. The same level of sera measles antibodies was observed in immunized people after intranasal or subcutaneous measles vaccination. Special investigation of measles vaccination side effects in adults revealed that the injected vaccine suppressed lymphocytes functional capacity much more than when intranasally introduced. Intranasal measles vaccine spray introduction may be a useful method of child revaccination in the process of measles eradication. This method is useful for investigation of "mucosal immunity" in children or adults. PMID- 10690627 TI - Characterization of monoclonal antibody CIBCNSH3 generated to the human EGF receptor. AB - Monoclonal antibody CIBCNSH3 of IgG1 isotype has been generated against human epidermal growth factor receptor (EGFR) using MDA MB 468 breast carcinoma cell line as immunogen. Earlier studies have revealed that this MAb blocked growth factor-receptor interaction and thus inhibited cell proliferation and tumor growth. In the present paper, this MAb has been extensively characterized to evaluate its application in the study of human cancers. The results were compared with those obtained using a control MAb ICR 62 specific to EGFR. Competitive assay showed that this MAb bound to an epitope in the extracellular domain of the EGFR to which MAb ICR 62 also bound. This MAb immunoprecipitated the 170 kD glycoprotein. The specificity was further confirmed by the formation of a single discrete band in western blot analysis. By flow cytometric analysis this monoclonal antibody revealed high binding affinity with MDA MB 468 cells. By immunocytochemical assay, out of 35 breast tumors studied, 40% were found to exhibit strong cell membrane staining and in the case of 25 oral cancers studied, 56% were strong positive. High expression of EGFR was observed in MDA MB 468 cells and HN 5 cells. These studies clearly indicate that MAb CIBCNSH3 might prove useful to identify tumors with high level of expression of EGFR associated with poor prognosis. PMID- 10690628 TI - Evaluation of the BT-1 serum assay for breast cancer. AB - The BT-1 assay which identifies a novel breast tumor associated serum analyte was performed for 143 patients previously diagnosed with breast cancer. Mucin tumor markers CA15-3/CA27-29 values were available for 50 patients and there was very minor overlap between patients positive by both tests. Patients' follow-up clinical status at sample draw was compared to BT-1 assay results. 27% of patients originally diagnosed as Stage II and 20% patients originally diagnosed as Stage III who were evaluated 'no disease' had positive BT-1 values. 8% patients diagnosed as Stage II had negative BT-1 results in samples drawn within 90 days of chemotherapy initiation, whereas 23% of patients diagnosed as Stage III cancer were BT-1 test positive within 90 days of chemotherapy initiation. 50% of patients tested before initial breast cancer surgery had positive BT-1 values, suggesting that the BT-1 assay may be useful in identification women with more advanced disease at diagnosis. PMID- 10690629 TI - Limited usefulness of antithyroperoxidase and antithyroglobulin assays in Jamaicans with Graves' disease. AB - The clinical usefulness of commercially prepared haemagglutination kits for thyroperoxidase (TPO) antibody and thyroglobulin (TG) antibody was evaluated in 145 consecutive Jamaicans with Graves' disease. Sixty two (43%) of the patients were newly diagnosed, 12 (8%) were in remission and 71 (49%) had relapsed. Sera from 65 (45%) patients were positive for thyroid antibodies. The TPO antibodies were found in 43% (63/145), thyroglobulin antibodies in 12% (17/145) fifteen (10%) patients had both anti-TPO and TG antibodies. Similar prevalences of TPO antibody were found in newly diagnosed patients and those who had relapsed (44% v. 41%) but the prevalence in the patients in remission was significantly higher (75%; X2 = 4.8, P < 0.05). The prevalence of TPO antibody increased significantly with age at onset before age 35 years compared to later onset (56% v. 32%; X2 = 8.09, P < 0.005). The presence of both TPO antibody (64% v. 31%; X2 = 13.1, P < 0.001) and TG antibody (22% v. 6%; X2 = 8.8, P < 0.005) correlated positively with Graves' ophthalmopathy. Neither of the tests was adequately sensitive in detecting GD in Jamaicans but we recommend testing for TPO antibody without TG antibody as a cost-effective approach. The presence and titres of TPO antibody and TG antibody do not correlate with disease activity and are not reliable enough for monitoring antithyroid drug therapy in GD. There is a need for antibody tests which are efficacious in diagnosing and monitoring antithyroid drug therapy in GD, and suitable for use in developing countries. PMID- 10690630 TI - From IgG monoclonals to IgM-like molecules. AB - One problem in blood group testing is that IgG monoclonal antibodies, in contrast to IgM, do not usually agglutinate erythrocytes. One of the reasons is the high zeta potential induced by the negative charge of the cell surface. During the last few years, we have produced a series of human monoclonal antibodies by the conventional fusion technique directed against antigens of the Rh blood group system. Some of these monoclonals, especially those directed against Rh-subgroups such as the c-antigen, were mainly of the IgG-subtype and unsuitable for agglutination tests. We have therefore tried to establish a molecular biological method to make IgM-like molecules from IgG monoclonals. From the c-antigen specific human hybridoma BS 240 (IgG subtype), we isolated mRNA that was transcribed into cDNA and then amplified by PCR using family specific primers. The heavy and light chain products were cloned into the pHen vector containing a DNA linker fragment, a myc-tag for identification and a His-tag for purification. After transformation in E.coli and phage rescue with helper phage, the culture supernatant was screened for antigen positive recombinant phage antibodies as a first control for specificity using c-antigen positive erythrocytes and anti-M13 antibodies as bridging antibodies (Coombs technique). Erythrocytes being negative for the c-antigen served as a negative control. After changing the culture conditions, soluble single chain fragments (scFv) were obtained from the periplasmatic extract. Specificity was shown using the c-antigen positive and negative erythrocytes and the 9E10 antibody (anti-myc) as a bridging antibody. To obtain IgM-like molecules, DNA coding for the specific scFv was cloned into the vector pSTE containing DNA coding for the monomer of core streptavidin. After expression, purification and refolding of the monomer, the core streptavidin combines to form tetrameric structures, termed scFv::strep, that are able to bind biotin as shown using ELISA plates coated with biotinylated BSA. Binding was detected with 9E10 and a peroxidase conjugated secondary antibody. In the agglutination assay, the construct was able to agglutinate c-antigen positive erythrocytes but not the negative erythrocytes. These experiments show that it is possible to construct IgM-like agglutinating molecules from cells containing secreting IgG antibodies. Experiments employing human antibody libraries instead of hybridoma cell lines are now in progress. PMID- 10690631 TI - Studies on p53 immunolocalisation in breast cancer and its prognostic significance. AB - Immunocytochemical localisation of mutant p53 in breast tumours serves as a potential prognostic molecular marker. In order to study the expression of p53 protein in breast cancer which constitutes the second most common malignancy in the South Indian female population, MAb CIBCVMC12 has been generated against human p53 protein isolated and purified from bacterial cell lysate of E.coli carrying the plasmid T 7-7 Hup53 grown in Luria broth to induce the expression of p53. The positive clones selected by ELISA were found to exhibit strong staining of nuclear p53 in both fresh and archival paraffin embedded breast tumour tissue sections. Commercial MAb D 07 against p53 was used as control. In immunoprecipitation, this MAb of IgG2b isotype was found to bind specifically to a protein of 53 kD. Immuno cyto chemical assay of normal, benign and malignant breast tissues of different histological types revealed that the majority of tumour cells were strong positive in the case of infiltrating ductal and lobular carcinomas, the staining being less intense for in situ carcinoma. The test for normal and benign tissues was negative. The staining patterns were comparable with those of control antibody. These results suggest that the MAb generated is specific to p53. The p53 protein expression was compared with the estrogen receptor (ER) status for 50 breast tumours which revealed that 38% of these were p53 positive and of these two were ER+. Among the p53 negative tumours, 48% were found to be ER+. A comparison of the p53 expression for 100 breast cancer patients indicated that 57% of the tumours were p53 negative and these patients had a longer overall 5 year survival rate and recurrent free interval which is statistically very significant. These results might suggest that p53 positive tumours are more aggressive biologically with poor prognosis. PMID- 10690632 TI - Analysis of VH and VL genes of a monospecific human anti-myosin antibody produced by a B cell from the primary repertoire. AB - Epstein-Barr virus (EBV) transformation of B lymphocytes from a Glanzmann's thrombasthenia patient with a serum antibody to the integrin alpha IIb beta 3, led to the immortalization of a B cell secreting a monospecific IgM monochonal antibody (MAb), B7, reactive with platelet myosin. Analysis of B7 V genes revealed minimally mutated sequences: the immortalized B cell issued from the primary repertoire, with no evidence of an in vivo selection by myosin. The V genes were here compared with sequences of human MAbs available on databases to more clearly understand the monospecificity of the B7 MAb. B7 V genes were closely identical to rearranged V genes in clones with self-specificities, often secreting polyreactive antibodies. In contrast, B7 is an unmutated monoreactive human MAb able to recognize myosin with a high avidity. Comparison of the CDR3H sequence with that of MAbs in databases supports a central role for the CDR3H subdomain in determining monospecificity. Our results suggest the existence of a monospecific autoreactive B cell compartment, besides the well-known polyspecific one, susceptible to be the template of pathogenic autoreactivity, characterized by antibodies of high affinity and specificity. PMID- 10690633 TI - Is the HemK family of putative S-adenosylmethionine-dependent methyltransferases a "missing" zeta subfamily of adenine methyltransferases? A hypothesis. AB - Previous comparative studies revealed close similarity among various groups of S adenosyl-L-methionine (AdoMet)-dependent methyltransferases (MTases), indicating their common evolutionary origin. We present evidence for a remarkable similarity between the sequence and predicted structure of HemK (a widespread family of putative proteins encoded in genomes from bacteria to humans) and the catalytic domain of the gamma-subfamily of adenine-specific DNA MTases (N6mA MTases). We predict the structure and function of the putative catalytic domain of HemK proteins and speculate that the target-recognizing function may be conferred by the N-terminal variable region. PMID- 10690634 TI - Speaking out of turn: a role for silent synapses in pain. AB - Severe tissue or nerve injury can result in a chronic and inappropriate sensation of pain, mediated in part by the sensitization of spinal dorsal horn neurons to input from primary afferent fibers. Synaptic transmission at primary afferent synapses is mainly glutamatergic. Although a functioning excitatory synapse contains both alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors in the postsynaptic membrane, recent evidence suggests that dorsal horn neurons contain some "silent" synapses, which exhibit purely NMDA receptor-mediated evoked postsynaptic currents and do not conduct signals at resting membrane potential. Serotonin, which is released onto dorsal horn neurons by descending fibers from the rostroventral medulla, potentiates sensory transmission by activating silent synapses on those neurons, i.e., by recruiting functional AMPA receptors to the postsynaptic membrane. This phenomenon may contribute to the hyperexcitability of dorsal horn neurons seen in chronic pain conditions. PMID- 10690635 TI - G beta gamma-mediated signaling: new therapeutic target for proliferative vascular disease. AB - Proliferation of vascular smooth muscle (VSM) severely affects the outcome of coronary artery bypass and angioplasty procedures, causing the failure of venous grafts or restenosis of the reopened vessel. Investigation into the mechanisms underlying the process of VSM cellular proliferation has provided evidence that intracellular signaling mechanisms triggered by extracellular hormonal factors acting through G protein-coupled receptors, can mediate and sustain this pathological process. Inhibition of common pathways of G protein-coupled receptor signaling has recently proven effective in preventing VSM cellular activation and proliferation. In particular, inhibition of the G beta gamma-mediated mitogen activated protein (MAP) kinase signaling pathway results in the inhibition of VSM proliferation in vitro. Moreover, use of adenoviral vectors to deliver a peptide inhibitor of G beta gamma signaling in vivo has resulted in inhibition of intimal hyperplasia in experimental models of vein-graft failure and restenosis. PMID- 10690636 TI - Insulin-induced oxidative neuronal injury in cortical culture: mediation by induced N-methyl-D-aspartate receptors. AB - While effectively attenuating neuronal apoptosis in mouse cortical culture, insulin paradoxically induced neuronal necrosis with 48 h of exposure. The insulin neurotoxicity was blocked by an antioxidant but not by caspase inhibitors. Exposure to insulin led to tyrosine phosphorylation of the insulin receptor and the insulin-like growth factor-1 (IGF-1) receptor and activation of protein kinase C (PKC) and phosphoinositide 3-kinase (PI3-kinase). Inhibitors of tyrosine kinase and PKC, but not PI3-kinase, attenuated the insulin neurotoxicity. Conversely, the inhibitor of PI3-kinase but not PKC reversed the antiapoptotic effect of insulin. Suggesting that the gene activity-dependent emergence of excitotoxicity contributed to insulin neurotoxicity, macromolecule synthesis inhibitors and N-methyl-D-aspartate (NMDA) antagonists blocked it. Consistently, exposure to insulin increased the level of the NR2A subunit of the NMDA receptor without much altering NR1 or NR2B levels. The present study suggests that insulin can be both neuroprotective and neurotoxic in the same cell system but by way of different signaling cascades. PMID- 10690637 TI - Direct transcriptional control of the chloroplast genes psbA and psaAB adjusts photosynthesis to light energy distribution in plants. AB - Two photosystems, I and II, absorb and convert light energy in photosynthesis in chloroplasts of green plants. The genes psbA and psaAB of the cytoplasmic chloroplast genome encode core components of photosystem II and photosystem I, respectively. Here we show that the absolute amounts of photosystem I and photosystem II respond, in a complementary manner, to changes in light quality that preferentially excite each photosystem in mustard seedlings. We also show that the initial response to altered energy distribution is a change in the rates of transcription of psbA and psaAB. Changes in chlorophyll fluorescence emission in vivo suggest that the signal initiating this change is the oxidation-reduction state of plastoquinone, a component of the photosynthetic electron transport chain that connects photosystem I and photosystem II. The results are consistent with transcriptional effects observed previously with chloroplasts isolated in vitro and demonstrate that redox control of chloroplast transcription initiates long-term adjustments that compensate for imbalance in energy distribution and adapt the whole plant to altered light environments. PMID- 10690638 TI - Identification of nuclear matrix and associated proteins that bind the haptoglobin gene cis-element. AB - To identify the major nuclear matrix proteins that bind to the rat haptoglobin gene cis-element, we isolated a soluble nuclear matrix protein fraction and analysed it by gel retardation. Two major DNA-binding proteins exhibiting different types of protein-DNA interactions were detected: a DNA sequence specific 32-kDa isoform of transcription factor C/EBP beta, and a nuclear matrix protein p55 that bound to the DNA nonspecifically. During increased transcription of the haptoglobin gene in the course of the acute-phase reaction, the DNA binding affinities and concentrations of these proteins in the soluble nuclear matrix fraction were increased. These data lend further evidence that the nuclear matrix is an active support structure that localizes gene regulatory proteins and participates in transcriptional regulation. PMID- 10690639 TI - Immunochemical determination of cellular content of translation release factor RF4 in Escherichia coli. AB - The biosynthesis of proteins in prokaryotes is terminated when a stop codon is present in the A-site of the 70S ribosomal complex. Four different translation termination factors are known to participate in the termination process. Release factor RF1 and RF2 are responsible for the recognition of the stop codons, and RF3 is known to accelerate the overall termination process. Release factor RF4 is a protein involved in the release of the mRNA and tRNA from the ribosomal complex. Furthermore, RF4 is involved in the proofreading in the elongation step of protein biosynthesis. The cellular contents of RF1, RF2, and RF3 were determined earlier. Here we report the cellular content of RF4 in Escherichia coli to be approximately 16,500 molecules per cell. The cells were grown in a rich medium and harvested in the beginning of the exponential growth phase. The quantifications were performed by using Western immunoblotting with radioactive iodinated streptavidin and biotinylated rabbit anti-mouse immunoglobulins plus a highly specific monoclonal antibody against RF4 as first antibody. PMID- 10690640 TI - Activation of the acidic isoform of phospholipase A2 from Naja mossambica mossambica venom by oleoyl imidazolide requires the cooperative action of two ionizing groups. AB - The acidic phospholipase A2 isoform from the spitting cobra Naja mossambica mossambica is activated irreversibly by treatment with a molar equivalent of oleoyl imidazolide. The kinetics of the chemical modification of the enzyme can also be monitored by measuring the large reduction of tryptophan fluorescence, which is accompanied by a distinct red shift. The addition of a single molar equivalent of oleic acid to the enzyme produces an instantaneous reduction in fluorescence but with a barely detectable red shift, confirming that the response to oleoyl imidazolide results from covalent modification of the protein rather than hydrolysis of the reagent. The pH dependence of both activation and fluorescence reduction by oleoyl imidazolide has an optimum rate near pH 8.0. We propose that long-chain fatty acids and long-chain acyl imidazolides bind at a single activation site and that the reaction of the imidazolides involves two protein residues, one of which is a nonessential histidine residue and the other a primary amino group. PMID- 10690641 TI - 5'-upstream structure of the gene coding for chicken riboflavin-binding protein and its relation to estrogen induction. AB - To elucidate the mechanism of the estrogen-dependent induction of chicken riboflavin-binding protein (RfBP), we analyzed the 5'-upstream structure of its gene. A noncoding exon exists there, and around this sequence, 9 widely spaced half-palindromic estrogen-response element (ERE) motifs (5'-GGTCA or 5'-TGACC) were found. Furthermore, an imperfect ERE-like palindromic sequence (5' ATGTCANNNTGACAT-3') was also found at the 2.25 kb upstream region. No consensus palindromic ERE was observed. By luciferase reporter assay, the regions containing the half ERE motifs and the imperfect ERE showed estrogen-dependent enhancer activities, suggesting that these two characteristic sequences might confer estrogen-inducibility upon the chicken RfBP gene. However the activities were lower than that of a consensus ERE. It remains uncertain whether these sequences act cooperatively. PMID- 10690642 TI - Existence of multiple novel Gs alpha splice variants in acute leukemia patients. AB - The alpha subunit of the stimulatory G protein, Gs alpha, is involved in stimulation of the adenylate cyclase pathway of signal transduction. In this study, we investigated the status of the Gs alpha gene in 29 acute leukemia patients and identified three novel splice variants (designated Gs alpha L-1, Gs alpha L-2, and Gs alpha L-3), possibly derived from aberrant splicing. All of the splice variants have in-frame deletions, removing the functional domain responsible for GTPase activity of Gs alpha, and would encode truncated proteins of 160(Gs alpha L-1), 90(Gs alpha L-2) and 70(Gs alpha L-3) amino acids, respectively. The data suggest that these novel products may be implicated in an as-yet-unidentified signal transduction pathway in hematopoietic cells. PMID- 10690643 TI - Rearrangement in the coding and 5' region of p53 gene in human oral tumors. AB - Different postreplicative, transcriptional, and translational mechanisms responsible for p53 gene inactivation are slowly unfolding. Rearrangement of the p53 gene is a very rare event in human solid tumors and has been reported only in osteosarcomas. From our laboratory we have recently reported rearrangement only in the coding region of the p53 gene in breast tumors. In this report we have undertaken a systemic investigation of p53 in oral tumors. Results have shown rearrangement in the coding region of p53 in 20% of cases and in the 5' region of p53 gene in approximately 8% of cases. No allelic loss and amplification of p53 gene was observed in these tumor samples. In our earlier studies on breast tumors, we found no abnormality in the 5' region of p53. However, in the present study we report for the first time rearrangement in the 5' region of p53 in oral tumors. Correlation of p53 gene rearrangement with p53 expression of RNA and protein indicates that rearrangement in the 5' region of p53 might not have a role in p53 expression. However, rearrangement in the coding region of p53 might play a critical role in controlling p53 gene activity in the process of tumorigenesis. PMID- 10690644 TI - Blockage of HIV-1 production through inhibition of proviral DNA synthesis by N,O didecanoyl serinal dimethylacetal. AB - Six serinal derivatives were synthesized and tested for their anti-human immunodeficiency virus type-1 (HIV-1) activity against HIV-1-infected cells. Of the 6 serinal derivatives tested, only N,O-didecanoyl serinal dimethylacetal (DDSD) was found to strongly suppress progeny virus production from acute HIV-1 infected CEM cells, while not suppressing the HIV-1 p24 production from latent HIV-1-infected ACH-2 cells after stimulation with phorbol 12-myristate 13 acetate. DDSD also inhibited the synthesis of HIV-1 proviral DNA at 20-50 microM, not only 1 h but also 24 h after HIV-1 infection. Taken together, DDSD is a potent inhibitor of HIV-1 production, and may become a unique leading compound for chemotherapy of acquired immunodeficiency syndrome. PMID- 10690645 TI - Real-time oligonucleotide hybridization kinetics monitored by resonant mirror technique. AB - The kinetics of hybridization of 11-meric and 14-meric oligonucleotides, dTGGGAAGAGGG (ODN-11) and dTGGGAAGAGG GTCA (ODN-14), with 14-meric oligonucleotide dpTGACCCTCT TCCCA (p14) attached to the surface of a cuvette was studied by the resonant mirror method. The treatment of the experimental curves with exponential equations leads to the following values for association (kas) and dissociation (kdis) rate constants at 25 degrees C: kas = 219 +/- 39 and 183 +/- 162 M-1 s-1, kdis = (2.0 +/- 0.4) x 10(-3) and (4 +/- 1) x 10(-4) s-1 for the duplexes (p14) x (ODN-11) and p14 x (ODN-14), respectively. The oligonucleotide dTGCCTTGAATGGGAA GAGGGTCA (ODN-23), which forms a hairpin structure, does not associate with p14. The data were compared with the results of melting curve detection and temperature-jump experiments. The association rate constants for ODN-11 and ODN-14 are much slower than those values in homogeneous aqueous solution. The dissociation rate constants have the same magnitude values as estimated by using association constants measured from melting curves but differ from the values estimated in temperature-jump experiments. PMID- 10690646 TI - Effect of rooperol on collagen synthesis and cell growth. AB - The effect of rooperol on type I collagen synthesis in normal skin and lung fibroblasts and cell growth in normal and transformed fibroblasts was investigated. Low concentrations of rooperol selectively inhibited the growth of transformed cells while stimulating collagen synthesis in normal fibroblasts. Elevated collagen synthesis and deposition could impede tumour cell invasion and metastasis, implying that rooperol may be useful as an antimetastatic agent in the treatment of cancer. PMID- 10690647 TI - Substitution of surface-exposed framework residues alters secretion of recombinant fusion protein Fv/tumor necrosis factor in Escherichia coli. AB - Substitution of hydrophobic residues with hydrophilic ones at surface-exposed positions may influence the yield of antibody fragment expression in Escherichia coli by reducing its aggregation potential. We introduced such substitutions at 8 surface-exposed framework region residues of a fusion protein Fv/Tumor Necrosis Factor, which resulted in the expression of 10 mutant fusion proteins (Mut 1-10) in E. coli. Our results showed that expression levels of Mut 1-3, with mutations of A9S, T18K, and G41D, respectively, decreased by 4- to 8-fold, whereas expression levels of Mut 4, 9, and 10, with mutations of S60D/S70D, T28D, and G65D, respectively, were not affected. In contrast, mutation of F83A at light chain residue 83, which is usually buried at the variable/constant domain interface of antibody molecules but becomes surface-exposed in recombinant Fv molecules, increased the expression level of Mut 5 by 4-fold. Our results suggest that this important substitution of a hydrophobic residue with a hydrophilic one may also be applied to increase the secretion of other recombinant Fv molecules in E. coli periplasm. PMID- 10690648 TI - Use of chimeric forms of neuronal nitric-oxide synthase as dominant negative mutants. AB - Because the functional form of neuronal nitric-oxide synthase (nNOS) is a homodimer, we investigated whether we could disrupt dimer formation with inactive nNOS chimeras acting as dominant negative mutants. To test this hypothesis, we either expressed the heme and reductase regions of rat nNOS as single domains or produced fusion proteins between the rat nNOS heme domain and various other electron-shuttling proteins. A dominant negative potential of these constructs was demonstrated by their ability to reduce NOS activity when transfected into a cell line stably expressing rat nNOS. In the presence of these nNOS mutant proteins, cellular levels of inactive nNOS monomers were significantly increased, indicating that their mechanism of action is through the disruption of nNOS dimer formation. These dominant negative mutants should prove valuable in analyzing the role of nNOS in biological systems. PMID- 10690649 TI - Biochemical properties of the protein tyrosine kinase of the bacterium Acinetobacter johnsonii. AB - The biochemical properties of the autophosphorylating protein tyrosine kinase of Acinetobacter johnsonii were analyzed in vitro. The study shows that the optimal pH value for the phosphorylation reaction is approximately 7. The enzyme activity is stimulated by magnesium and, to a lesser extent, by manganese ions, whereas calcium ions have no effect. The phosphorylation process is rapid reaching a maximum in < 2 min, and the enzyme is modified at multiple sites. Interestingly, the bacterial enzyme is insensitive to a series of molecules known to affect the activity of eukaryotic protein tyrosine kinases: genistein, quercetin, tosyllysine chloromethyl ketone, and vanadate. We concluded that, even though the overall phosphorylation reaction catalyzed by the A. johnsonii enzyme is identical to that occurring in eukaryotes, this bacterial kinase exhibits a number of specific properties and therefore probably belongs to a separate group in the general family of protein tyrosine kinases. PMID- 10690650 TI - Cloning and characterization of manganese superoxide dismutase gene from Vibrio parahaemolyticus and application to preliminary identification of Vibrio strains. AB - The sodA gene coding for manganese superoxide dismutase (Mn-SOD) from the marine microorganism Vibrio parahaemolyticus was cloned, sequenced, and overexpressed in Escherichia coli by use of the pET20b (+) expression vector. The full-length gene consisted of a 588-bp open reading frame and encoded a polypeptide of 196 amino acid residues, with a calculated molecular mass of 21,713 Da. The recombinant enzyme was efficiently purified from crude E. coli cell lysate by metal ion affinity chromatography. The recombinant VPMn-SOD resisted thermo-denaturation up to 60 degrees C and was insensitive to such inhibitors as EDTA, NaN3 and diethyldithiocarbamic acid. The specificity of V. parahaemolyticus Mn-SOD gene probe was analyzed by cross-species polymerase chain reaction to provide information for Vibrio strain identification. PMID- 10690651 TI - Characterization of cytostatically active glycosphingolipids isolated from thioglycollate-elicited murine macrophages. AB - Two acidic glycosphingolipids (gangliosides) derived from mouse macrophage membranes and separated by thin-layer chromatography have a strong cytostatic effect on human and mouse tumor cells. The structure of the two gangliosides, named M phi G1 and M phi G2, was elucidated by application of physicochemical and immunochemical methods. Gas chromatography and mass spectrometry of M phi G1 and M phi G2 classified them as isomeric monosialogangliosides with ceramide moieties composed of sphingosine as the long-chain base, C16 and C18 fatty acids, respectively, and a lacto-tetraose backbone. For M phi G1, additional immunochemical findings led to the proposed structure IV3NeuAc-nLcOse4Cer. The immunochemical reactions of M phi G2 suggest a branched structure for the oligosaccharide moiety. PMID- 10690652 TI - Production of superoxide and nitric oxide by granulocytes in non-insulin dependent diabetic patients with and without diabetic nephropathy. AB - Production of the superoxide radical anion O2-. and the nitric oxide radical NO-. by granulocytes was studied in 14 patients with type 2 diabetes without nephropathy, 21 patients with type 2 diabetes and diabetic nephropathy, and 19 healthy subjects, both without and after stimulation with opsonized zymosan. O2-. production by both resting and stimulated granulocytes was increased in type 2 diabetes patients without nephropathy but decreased in type 2 diabetes patients with nephropathy, compared with healthy subjects. NO. generation was highly augmented in type 2 diabetes patients without nephropathy by both resting and stimulated cells; values for type 2 diabetes patients with nephropathy were intermediate between the type 2 diabetes patients without nephropathy and the healthy subjects. These data point to granulocytes as one of possible sources of oxidative stress in type 2 diabetes. PMID- 10690653 TI - Failure to attract and retain clinician/scientist faculty puts our profession at risk. PMID- 10690654 TI - The road to preventive dentistry--the personal scientific experience of a Japanese dentist. PMID- 10690655 TI - Her name is "Lucy", our three-million-year-old ancestor. AB - Dental anthropology is a key discipline in studies to determine the evolutionary history of our hominid ancestors, to identify the origin and dispersal of modern humans, and to reconstruct the source of observed dental variation. A survey of hominid and modern human evolutionary history, emphasizing results from powerful multivariate dental morphometric methodologies, suggests a single African origin of modern humans > 150,000 years before present from a Homo heidelbergensis ancestor. A continuum among modern humanity is described, with, first, sub Saharan Africans, then southeast Asian Negrito, and Australian aborigines at its extant root. Other interpretations of the available data are possible. Examinations of the progress of the evolution of teeth through time give significant insight into dental morphogenetics and variation, and the biology of dental evolution. The mechanisms of evolution which fashion a phenotype and the methods of molecular and dental phylogenetics are reviewed and evaluated. This is an exciting time for dental anthropology, with fascinating and challenging questions to address, but anthropologists, not dentists, dominate the field. The perspective of a dentist can meaningfully add to the dynamics of dental anthropology. PMID- 10690656 TI - Architecture of intact natural human plaque biofilms studied by confocal laser scanning microscopy. AB - Determination of the structure of human plaque will be of great benefit in the prediction of its formation and also the effects of treatment. However, a problem lies in the harvesting of undisturbed intact plaque samples from human volunteers and the viewing of the biofilms in their natural state. In this study, we used an in situ device for the in vivo generation of intact dental plaque biofilms on natural tooth surfaces in human subjects. Two devices were placed in the mouths of each of eight healthy volunteers and left to generate biofilm for 4 days. Immediately upon removal from the mouth, the intact, undisturbed biofilms were imaged by the non-invasive technique of confocal microscopy in both reflected light and fluorescence mode. Depth measurements indicated that the plaque formed in the devices was thicker round the edges at the enamel/nylon junction (range = 75-220 microm) than in the center of the devices (range = 35-215 microm). The reflected-light confocal images showed a heterogeneous structure in all of the plaque biofilms examined; channels and voids were clearly visible. This is in contrast to images generated previously by electron microscopy, suggesting a more compact structure. Staining of the biofilms with fluorescein in conjunction with fluorescence imaging suggested that the voids were fluid-filled. This more open architecture is consistent with recent models of biofilm structure from other habitats and has important implications for the delivery of therapeutics to desired targets within the plaque. PMID- 10690657 TI - Electron microscopy, micro-analysis, and X-ray diffraction characterization of the mineral-like tissue deposited by human cementum tumor-derived cells. AB - The nature and characteristics of the mineralized-like tissue deposited by cementoblasts are not well-known due to the difficulties in obtaining and culturing cells representing the cementum phenotype. We hypothesized that a putative cementoblastic cell line derived from a human cementoblastoma could serve as a suitable model to study the physical, chemical, and morphological features of the cementum-like tissue deposited in vitro. The cementoblastoma cell line was studied by transmission electron, high resolution, scanning, and atomic force microscopy and compared with human cellular cementum, human osteoblasts, and human alveolar bone. The analyses of the crystals and mineral-like tissue in the cell line were performed by x-ray diffraction microscopy and energy dispersive x-ray micro-analysis. TEM examination of cementoblastoma cells revealed the presence of electron-dense intracellular vesicles surrounded by a membrane that contained filaments and needle-like structures. The diffraction patterns obtained from the intracellular material and human cellular cementum were similar, with D-spacings of 3.36 and 2.8, consistent with those of hydroxyapatite (3.440 and 2.814). The composition of the mineral-like tissue had a Ca/P ratio of 1.60 for cementoblastoma cells and 1.97 for human cellular cementum. Na (5.29%) and Cl (1.47%) were present in the composition of cementoblastoma cells. Human cellular cementum additionally contained Mg (4.95%). Osteoblastic cells showed a Ca/P ratio of 1.6280. Na represented 4.52% and Cl 1.22% of its composition. Human alveolar bone had a Ca/P ratio value of 2.01. Na (6.63%), Mg (2.10%), and Cl (0.84%) were also present. All samples examined represented biological-type hydroxyapatite. Based on the compositional and morphological features, these findings indicate that cementoblastoma-derived cells express the human cellular cementum phenotype. PMID- 10690658 TI - Quantification of periapical bone destruction in mice by micro-computed tomography. AB - Bacterial infections of the dental pulp result in tissue destruction and periapical bone resorption. The availability of genetically engineered mouse strains is a major advantage in the use of this model system for studies of periapical pathogenesis. The main limitation of the mouse model is its small size, and the necessity for laborious histologic analyses to quantify periapical bone destruction. In the present study, we evaluated the use of a new technology, high-resolution micro-computed tomography (micro-CT), for the rapid and non invasive quantification of periapical bone destruction. Periapical lesions were induced in the lower first molars of mice by exposing the pulp to the oral environment. Mandibles were harvested on day 21 after pulp exposure, and were subjected to micro-CT analysis, with 17-microm-thick radiographic sections. Samples were then decalcified, embedded, and sectioned for histology. The cross sectional area of periapical lesions was determined by image analysis of corresponding micro-CT and histologic sections. The results showed a highly significant correlation between micro-CT and histology (p < 0.0001), with mean differences of 4. 1% (range, 0.9 to 7.2%) between the two methods. The mean error associated with image analysis was 4.9% for images obtained by both micro-CT and histology. The variability of replicate (n = 5) independent micro-CT determinations was 3.4%, less than that associated with the image analysis error. These results demonstrate that micro-CT imaging is a rapid, reproducible, and non invasive method, that gives results that are closely comparable with those obtained by histology. Micro-CT appears to have utility for the accurate quantification of changes in bone architecture in small biological specimens. PMID- 10690659 TI - Nominal shear or fracture mechanics in the assessment of composite-dentin adhesion? AB - This study addresses the anticipated problem of discriminating among high performing dentin adhesives. The simplicity of the nominal shear bond test, despite being heavily criticized, has made it a routine procedure for the determination of bonding efficacy. A fracture mechanics approach has been suggested as a better assessment of bonding efficacy (Versluis et al., 1997). However, experimental complexity is a major limitation. It is hypothesized that a new, simplified interfacial fracture toughness test (Lin, 1994) will evaluate bonding agents differently if compared with the traditional shear bond test. Therefore, the objective of this study was to compare the performances of six dentin bonding agents subjected to the interfacial fracture toughness test (critical plane strain energy release rate) or to the nominal shear bond test (shear bond strength). Their performances were also characterized by scanning electron micrography of the fracture surfaces for evidence of dentin cohesive failure. Statistical analyses showed only marginal differences between these determinants of the two tests. However, when the analysis was applied only to the materials that had 100% frequency of dentin cohesive failure in shear testing, which also had high bonding efficacy, the difference in adhesive strengths between the two tests became significant. The reliability of the nominal shear test is questioned when dentin cohesive failure occurs, which usually is associated with high bonding efficacy. Since it is expected that bonding efficacy will increase further, the interfacial fracture toughness test is the preferred methodology to distinguish among high-performing dentin adhesives. PMID- 10690660 TI - Acute-phase inflammatory response to periodontal disease in the US population. AB - Moderate elevation of serum C-reactive protein (CRP) is a risk factor for cardiovascular disease among apparently healthy individuals, although factors that create this inflammatory response in the absence of systemic illness have not been clarified. This study aimed to: (1) evaluate associations among periodontal disease, established risk factors for elevated CRP, and CRP levels within the US population; and (2) determine whether total tooth loss is associated with reduced CRP. Data were obtained from the third National Health and Nutrition Examination Survey. A random sample of the US population was interviewed in their homes and examined at mobile examination centers. CRP was quantified from peripheral blood samples and analyzed as a continuous variable and as the prevalence of elevated CRP (> or = 10 mg/L). Some 12,949 people aged 18+ years who had periodontal examinations and an additional 1,817 edentulous people aged 18+ years were included in the analysis. Dentate people with extensive periodontal disease (> 10% of sites with periodontal pockets 4+ mm) had an increase of approximately one-third in mean CRP and a doubling in prevalence of elevated CRP compared with periodontally healthy people. Raised CRP levels among people with extensive periodontal disease persisted in multivariate analyses (P < 0.01), with established risk factors for elevated CRP (diabetes, arthritis, emphysema, smoking, and anti-inflammatory medications) and sociodemographic factors controlled for. However, CRP levels were similarly raised in edentulous people. Furthermore, the established risk factors for elevated CRP modified relationships between oral status and CRP levels. Periodontal disease and edentulism were associated with systemic inflammatory response in the US population, most notably among people who had no established risk factors for elevated CRP. PMID- 10690662 TI - A controlled study of the morphometric changes in the primary dentition of pre term, very-low-birthweight children. AB - The aim of the present investigation was to compare the dimensions of the primary incisors from pre-term children and full-term controls. One hundred and eleven pre-term children, consisting of 86 very-low-birthweight (< 1,500 g), 25 low birthweight (from 1,500 to 2,500 g), and 169 full-term, normal-birthweight (> 2,500 g) children, donated a total of 572 maxillary and mandibular primary central and lateral incisors for study. The teeth were measured by means of a digital micrometer. The results showed that there was a dose-response effect of birthweight on tooth size. The very-low-birthweight teeth showed the smallest dimensions, the normal-birthweight controls the largest, and the low-birthweight teeth intermediate dimensions (p < 0.001). In the maxillary primary central and lateral incisors, and the mandibular primary central incisors, very-low birthweight teeth were from 6 to 11% smaller in both mesiodistal and faciolingual dimensions compared with normal-birthweight teeth (p < 0.001). The largest differences were observed in the maxillary lateral incisors, where mean decreases of 0.58 +/- 0.45 mm in mesiodistal and 0.50 +/- 0.40 mm in faciolingual dimensions (11% reduction in both measurements) were observed. In addition, in the mandibular and maxillary lateral incisors of very-low-birthweight children, the left-sided teeth were significantly smaller than those on the right side in both mesiodistal and faciolingual dimensions (p < 0.03). PMID- 10690661 TI - Composition of plaque and saliva following use of an alpha-tricalcium-phosphate containing chewing gum and a subsequent sucrose challenge. AB - Previous studies demonstrated that the chewing of a 2.5% (mass fraction) alpha tricalcium-phosphate-fortified (alpha-TCP) experimental chewing gum released sufficient calcium and phosphate to eliminate any fall in the tooth mineral saturation of plaque fluid after a sucrose rinse (Vogel et al., 1998). In contrast, the chewing of a conventional sugar-free gum did not eliminate this decrease in saturation. The purpose of this study was to examine if the release of ions from plaque calcium-phosphate pools induced by this gum could provide protection during subsequent exposure to cariogenic conditions. Fourteen subjects accumulated plaque for 48 hrs, fasted overnight, chewed a control or experimental gum for 15 min, and subsequently rinsed 1 min with a mass fraction 10% sucrose solution. Before gum chewing, and at 7 min and 15 min afterward, whole plaque, plaque fluid, and salivary samples were obtained and analyzed by micro-analytical techniques. Additional samples were collected and analyzed at 25 min (7 min after the sucrose rinse). Although the results confirmed the deposition of large amounts of calcium and phosphates in plaque seen in the previous study, only a small increase was seen in plaque-fluid-free calcium and phosphate before sucrose administration. This suggests that few of the mineral ions were mobilized under non-cariogenic conditions. However, 7 min after the sucrose rinsing, an increase in these concentrations was seen which, based on hydroxyapatite ion activity product calculations, indicated a decrease in the driving force for demineralization compared with that seen with the control gum. These results suggest that the chewing of the experimental gum deposits a labile mineral reservoir in plaque that can resist a subsequent cariogenic challenge. PMID- 10690664 TI - The expression of MMP-8 in human odontoblasts and dental pulp cells is down regulated by TGF-beta1. AB - Recent findings show that matrix metalloproteinase-8 (MMP-8) is expressed, in addition to neutrophils, by human chondrocytes, cultured fibroblasts, and endothelial cells. We investigated the expression of MMP-8 in other human mesenchyme-derived cells, odontoblasts, and pulp tissue. Odontoblasts and pulp tissue were collected from extracted human teeth for MMP-8 mRNA analysis with reverse-transcription/polymerase chain-reaction (RT-PCR) and Southern blot. The expression, localization, and secretion of MMP-8 protein were studied with Western blot, immunohistochemistry, and immunofluorometric assay. The effect of TGF-beta1 (10 ng/mL) on the expression, secretion, and concentration of secreted MMP-8 was studied by odontoblast and pulp tissue culture methods (Tjaderhane et al., 1998a). RT-PCR demonstrated MMP-8 mRNA expression in native and cultured odontoblasts and pulp tissue and cultured pulp fibroblasts, with a 522-bp transcript comparable with that of bone marrow cells. The specificity of PCR was confirmed with Southern blot. Western blot with MMP-8-specific antibody detected 65- and 50-kDa proteins in native samples, representing latent and active forms of mesenchymal-type MMP-8, and in the conditioned odontoblast culture media, 50 kDa protein was observed. TGF-beta down-regulated the MMP-8 mRNA and concentration of secreted protein in both cultures. Immunohistochemical staining detected MMP-8 in odontoblasts. These findings indicate that mesenchyme-derived cells of the dentin-pulp complex express, synthesize, and activate MMP-8, which may, in concert with odontoblast-derived gelatinases, participate in organization of dentin organic matrix prior to mineralization. PMID- 10690663 TI - Localization of EMSP1 expression during tooth formation and cloning of mouse cDNA. AB - Enamel matrix serine proteinase 1 (EMSP1) is a proteolytic enzyme that has been isolated from the developing enamel of pig teeth. Its apparent function is to degrade the organic matrix in preparation for enamel maturation. The expression of EMSP1 has never been investigated in another organism besides the pig, and EMSP1 expression in the enamel organ has never been specifically demonstrated in ameloblasts. Here we report the expression of recombinant pig EMSP 1 (rpEMSP 1), the generation of rabbit polyclonal antibodies against rpEMSP1, the characterization of the antibodies and EMSP1 expression by Western blot and immunohistochemical analyses, the cloning and characterization of a full-length cDNA encoding mouse EMSP1, and the localization of EMSP1 expression in ameloblasts in mouse day 14 first and second molars by in situ hybridization. The full-length mouse EMSP1 cDNA clone has 1,237 nucleotides, excluding the poly(A+) tail, and encodes a preproprotein of 255 amino acids. Mouse EMSP1 shares 75% amino acid identity with pig EMSP1 and has three potential N-linked glycosylation sites, two of which are conserved in the pig homologue. Western blot analysis shows that the polyclonal antibodies are specific for EMSP1 and do not cross react with trypsin. Immunohistochemistry of pig incisors shows discrete staining in the surface enamel at the earliest part of the maturation stage. In mouse molars, in situ hybridization gives a distinct and specific signal in maturation stage ameloblasts, and in the junctional epithelium following tooth eruption. We conclude that EMSP1 is expressed by pig and mouse ameloblasts during the early maturation stage of amelogenesis. PMID- 10690665 TI - Regional differences of metabolism in human masseter muscle by two-dimensional 31P-chemical shift imaging. AB - Many reports have demonstrated significant region-dependent differences in the fiber-type composition of the human masseter muscle. Therefore, it is considered that there is intramuscular heterogeneity of metabolic activity in the muscle. The present study was carried out, with two-dimensional Chemical Shift Imaging, to detect differences between the deep and superficial parts of the human masseter muscle at rest. Masseter muscle from 11 volunteers, from 20 to 27 years old, was examined, and characteristic spectra of the inorganic phosphate (Pi), phosphocreatine (PCr), and alpha-, beta-, and gamma-adenosine triphosphate (ATP) from each part of the muscles were obtained. In this study, the deep and superficial parts of the masseter muscle were distinguished by the existence of aponeurosis. The Pi/PCr, PCr/beta-ATP, and Pi/beta-ATP ratios as well as the pH in the deep and superficial parts were calculated from the peak spectra. Compared with the deep part, the Pi/PCr of the superficial part was lower (p < 0.05) and the PCr/beta-ATP was higher (p < 0.01). The Pi/beta-ATP and pH showed no significant differences between the two parts. The results indicate that the superficial part of the masseter muscle contains more PCr than the deep part, and this may be related to functional differences between these two parts. In future examinations of the metabolic activity of the human masseter muscle, the deep and superficial parts must be measured separately. PMID- 10690666 TI - Effects of organic acid anions on growth, glycolysis, and intracellular pH of oral streptococci. AB - Oral streptococci produce large quantities of organic acids as the end-products of carbohydrate fermentation. In an approach to determine if oral streptococci exhibit differential sensitivities to organic acid anions, we determined the effects of formate, lactate, and acetate on intracellular pH maintenance, glycolysis, and growth of Streptococcus mutans and Streptococcus sanguis. Growth was determined as maximum culture optical density in the presence of the organic acid anions at pH 7.1, 6.7, 6.3, and 6.1, and the effects of the anions on glycolytic activity and intracellular pH were determined at pH 7.0 and 5.0. At pH 7.1, the organic acid anions had little effect on growth of either species. At the lower pH values, all of the anions reduced the maximum culture optical density of both species in a pH- and concentration-dependent manner, with S. sanguis being more sensitive to growth inhibition than S. mutans. The organic acid anions had little or no effect on glycolytic activity of either species at pH 7.0. However, all of the organic acid anions tested reduced glycolytic activity at pH 5.0 in a concentration-dependent manner, with S. sanguis being more sensitive than S. mutans. The inhibition of glycolysis could be related to the pKa of the organic acid, with formate and lactate being more inhibitory than acetate. The organic acid anions decreased the intracellular pH of S. mutans and S. sanguis, glycolyzing at an extracellular pH of 5.0, such that the reduction in glycolytic activity caused by the organic acid anions could be directly attributed to the fall in intracellular pH. In conclusion, the production of lactic acid in plaque would not only lower pH, thereby having a disadvantageous effect on less aciduric oral streptococci, such as S. sanguis, but would also increase their sensitivity to the effects of low pH, helping S. mutans to become more dominant. PMID- 10690667 TI - Is laparoscopy the gold standard for the diagnosis of endometriosis? PMID- 10690668 TI - Leptin, the ob gene product, in female health and disease. AB - Leptin is a recently discovered hormone which is involved in the regulation of body weight. It provides a molecular basis for the lipostatic theory of the regulation of energy balance. White adipose tissue is the main site of leptin synthesis and there is some evidence of ob gene expression in brown fat. Leptin seems to play a key role in the control of body fat stores by coordinated regulation of feeding behaviour, metabolic rate, autonomic nervous system regulation and body energy balance in rodents, primates and humans. Apart from the function of leptin in the central nervous system on the regulation of energy balance, it may well be one of the hormonal factors that signal the body's readiness for sexual maturation and reproduction to the brain. During late pregnancy and at birth when maternal fat stores have been developed leptin levels are high. Leptin could then be a messenger molecule signaling the adequacy of the fat stores for reproduction and maintenance of pregnancy. At later stages of gestation leptin could signal the expansion of fat stores in order to prepare the expectant mother for the energy requirements of full term gestation, labour and lactation. This overview focuses on those topics of leptin research which are of particular interest in reproductive medicine and gynecology. PMID- 10690669 TI - Appendix perforation by an intrauterine contraceptive device. AB - Perforation of the uterus by an intrauterine contraceptive device (IUD) is a rare, and serious complication, occurring in 1/350 to 1/2500 insertions. Perforation by IUDs can involve several neighboring organs such as the bladder and rectosigmoid. We report two cases of IUD perforations involving the appendix, both inserted during lactation. The first case is an asymptomatic patient in early pregnancy and the second is a woman whose original presentation was chronic lower abdominal pain. The presence of copper in the abdominal cavity can lead to adhesion formation and subsequent abdominal pain, bowel obstruction or infertility. Thus, we believe that when an IUD is located in the abdominal cavity it should be removed even in an asymptomatic patient. In addition, these cases might suggest postponing the use of this contraceptive method in lactating women. PMID- 10690670 TI - Obstetric brachial plexus injury: risk factors related to recovery. AB - OBJECTIVE: To investigate if multivariate risk calculation can discriminate those infants who do not recover after an obstetric brachial plexus injury (OBPI). STUDY DESIGN: All liveborn infants without lethal congenital abnormalities from 1988 through 1996 with a gestational age > or =30 weeks were included. Outcome variables were all OBPI and non-recovered OBPI. Risk calculation was performed by univariate analysis for all infants and by multivariate logistic analysis for all singleton infants delivered vaginally in cephalic presentation. RESULTS: A total of 62 of 13 366 liveborn infants sustained an OBPI (0.46%). Seventeen (27%) did not recover completely. Birth weight, female sex, second stage >60 min, diabetes, multiparity, maternal age and non-Caucasian origin were important risk factors for non-recovered OBPI. A model without birth weight, which can not be measured accurately antepartum, is considerably less effective. Risk factors for all OBPI and for non-recovered OBPI were similar. CONCLUSION: A predictive multivariate model is of limited value due to the low incidence of non-recovered OBPI. However, it may be useful to discriminate individual cases with exceptional risk. PMID- 10690671 TI - Preliminary experience with twenty perineal repairs using Indermil tissue adhesive. AB - OBJECTIVE: To assess the use of Indermil tissue adhesive for perineal repair. SETTING: Leeds General Infirmary, a teaching hospital with 4500 deliveries annually. METHOD: Over a period of five months, 20 women who sustained either a second degree tear or who had an episiotomy at vaginal delivery had their perineal skin repaired with Indermil tissue adhesive. They were followed up prior to discharge and then by telephone once discharged. RESULTS: Ten repairs followed normal vaginal deliveries, six were after ventouse deliveries, three after midcavity forceps delivery and one after a rotational forceps delivery. Three women noticed a burning sensation during application of adhesive. At follow up, 13 women were completely without problems, two complained of a sharp sensation from excess adhesive, one had silver nitrate applied at the six week check, two had small defects in the skin which healed well and in two women the skin edges broke down completely but did not need resuturing. CONCLUSION: Indermil tissue adhesive appears to be a safe and effective method of skin closure for episiotomies and second degree tears. The skin closure is quick and painless. PMID- 10690672 TI - Diclofenac in the treatment of pain after caesarean delivery. An opioid-saving strategy. AB - OBJECTIVE: Pain relief of good quality after caesarean section (CS) results in early mobilization and good early mother-child interaction. Patient-controlled analgesia (PCA), with systemic opioids, gives a very high level of patient satisfaction. However, opioids have well documented side-effects i.e. sedation, nausea and respiratory depression. To minimize the risk of such negative effects we studied how far the required dose of opioid could be decreased with a multimodal strategy adding diclofenac. STUDY DESIGN: In a randomized double-blind study, 50 parturients scheduled for elective CS under spinal anaesthesia, received rectally either diclofenac (Suppositorium diclofenac) 50 mgx3 or placebo 1x3 during the first 24 h postoperatively. All patients had PCA with the possibility of self-administered doses of ketobemidone 1 mg/6 min. RESULTS: In the group receiving diclofenac rectally the consumption of ketobemidone was reduced with 39% compared to the placebo group. CONCLUSION: A multimodal analgetic strategy with the addition of 150 mg diclofenac during the first 24 h after CS reduces the need for opioids significantly with maintained or improved analgetic effect. This is expected to reduce the risk of negative side-effects of systemic opioids. PMID- 10690673 TI - Maternal neonatal outcome in quadruplet and quintuplet versus triplet gestations. AB - OBJECTIVE: Examination and comparison of the natural histories of triplet versus quadruplet and quintuplet gestations. STUDY DESIGN: A retrospective study of sixty-four multifetal pregnancies (fifty-two sets of triplets, nine sets of quadruplets and three sets of quintuplets) cared for during past 12 years in our department. Quintuplets and quadruplets were compared with triplet pregnancies according to gestational age, birthweight, pregnancy complications and perinatal outcome. Student's t-test, Fisher exact test and chi2 test were used for statistical analysis, considering P value of <0.05 as statistically significant. RESULTS: Although mean gestational age at delivery between triplets and higher order gestations was not significantly different, birthweight of quadruplets and quintuplets was significantly lower. Pregnancy complications, including intrauterine growth retardation, were equally distributed between the groups. Early neonatal and perinatal mortality were significantly higher in quadruplets and quintuplets than in triplets. Surprisingly, survival of growth retarded fetuses was better than survival of their eutrophic counterparts. The spontaneous loss rate was 11.5% for entire triplet gestation and 16.7% for quadru- and quintuplet pregnancies. CONCLUSIONS: As the spontaneous loss rate of triplets and higher order pregnancies observed in our study is quite similar to pregnancy loss rate caused by multifetal pregnancy reduction, conservative management of multifetal pregnancies in specialised tertiary centers seems to be a prudent solution. PMID- 10690674 TI - Mature cystic teratomas of the ovary: case series from one institution over 34 years. AB - OBJECTIVE: To evaluate bilaterality, complications and malignant changes of mature cystic teratomas of the ovary. STUDY DESIGN: Retrospective study of 501 patients operated at Hacettepe University Hospital between the years of 1964 and 1998. RESULTS: The median age was 35 years (range 13-76). One hundred and six cases (21.1%) were asymptomatic. The mean tumor diameter was 7.0+/-4.5 cm. The decision for cystectomy or oophorectomy was related with the patient age, gravidity and parity. The bilaterality rate when both ovaries were evaluated histopathologically was 13.2% (44/331). Total complication rate was 10.7%, torsion being the most frequent (4.9%). The rate of malignant transformation was 1.4%. CONCLUSION: Ovarian mature cystic teratomas are common tumors especially during the reproductive period with low rates of covert bilaterality, complications and malignant transformation. The treatment should be directed on the basis of age, fertility desire or presence of another pelvic pathology rather than the size or bilaterality. PMID- 10690675 TI - Role of appendectomy in predicting lymph node metastases in patients with ovarian cancer. AB - OBJECTIVE: The aim of this study was to assess the diagnostic accuracy of appendectomy in predicting lymph node metastases in women undergoing cytoreductive procedures for ovarian cancer. STUDY DESIGN: In 127 consecutive patients with ovarian carcinoma appendectomy was performed in 30 patients over a period of 5 years. Eight of them were found to have metastases to the appendix. Pelvic and paraaortic lymphadenectomy was performed in 34 patients, in 24 of them the appendix was removed during primary surgery. RESULTS: Among 19 patients without metastases to the appendix the lymph nodes were positive in five cases (26.3%) and among five patients with metastases to the appendix the lymph nodes were positive in four cases (80.0%), which is not a significant difference. Evaluation of the appendiceal metastases as a predictor of lymph node metastases in patients with ovarian cancer gives a sensitivity of 44%, a specificity of 93%, a positive predictive value of 80%, a negative predictive value of 74% and an accuracy of 75%. CONCLUSION: The possibility of predicting retroperitoneal lymph node metastases in ovarian cancer on the basis of histological examination of the appendix is limited. PMID- 10690676 TI - Inhibitory G protein alpha subunit (Gi alpha) expression and localization during human trophoblast differentiation. AB - OBJECTIVE: Cyclic adenosine monophosphate (cAMP) participates in the regulation of processes associated with trophoblast syncytialization. As guanosine triphosphate (GTP)-binding regulatory proteins (G-proteins) modulate adenylate cyclase activity, the present study investigated the expression and regulation of the alpha subunit of inhibitory G protein (Gi alpha) during human trophoblast differentiation in vitro. STUDY DESIGN: Protein levels and the immunolocalization of the protein at a subcellular-level were assessed. RESULTS: Expression of Gi alpha protein decreased during syncytialization. Moreover, Gi alpha transmigrated from a predominantly cell membrane-associated to a predominantly perinuclear location during the process. CONCLUSION: These results suggest that Gi alpha is regulated and may be implicated in cAMP-dependent events during the terminal differentiation of human trophoblasts. PMID- 10690677 TI - Two month glucocorticoid treatment increases estradiol-induced stromal and myometrial cell proliferation in the uterus of ovariectomized rats. AB - The present work was undertaken to examine the effect of 2 month glucocorticoid treatment on estradiol-induced proliferation in the uterus. Ovariectomized rats were treated with long-acting triamcinolone acetonide or saline for 60 days followed by a single injection of estradiol dipropionate or vehicle. Proliferation in the uterus was estimated by the mitotic index, bromodeoxyuridine labelling index and proliferating cells nuclear antigen-labelling index 24, 36 and 48 h after the injection of estradiol or vehicle. Two month glucocorticoid treatment resulted in an initial decrease in all the parameters for luminal and glandular epithelia followed by an increase to 48 h after estradiol injection, and in a large increase in all proliferative parameters for stromal and myometrial cells at all periods of observation, compared with control rats untreated with glucocorticoid. The 2 month glucocorticoid treatment protocol had no significant effect on the parameters without estradiol administration. Analysis of proliferation for myometrial cells was also performed after 1 month treatment with glucocorticoid followed by estradiol or saline. Results showed no significant influence of 1 month treatment with glucocorticoid on myometrial cells. PMID- 10690678 TI - Significance of chromosomal aberrations for the unsuccessful procedures of assisted reproduction. AB - Lymphocyte cultures from patients who had previously undergone at least three unsuccessful procedures of assisted reproduction were analysed for cytogenetic abnormalities. A total of 12,657 metaphases from 33 individuals (15 patients and 18 healthy controls with two normal offsprings) were studied. A significantly higher incidence (P<0.001) of chromosome aberrations was found in the patients (6.79+/-0.68%; x+/-SD) as compared to the controls (1.72+/-0.3%; P<0.001). Chromosomal breakages, particularly at the centromere region, were also observed with significantly increased frequency in the patients than in the controls (6.18+/-0.65 vs. 1.42+/-0.27%, respectively; P<0.001). It is possible that the high rate of centromere breakages in the ART patients (3.18+/-0.47 vs. 0.26+/ 0.12%, P<0.001) may predispose to meiotic chromosomal abnormalities. A single cell aberration was demonstrated in 0.61+/-0.21% of ART patients' lymphocytes versus 0.3+/-0.12% in the controls (P<0.01). Structural rearrangements and chromosomal breaks predominantly affected the bands containing genes for the immune response and the cell cycle. Mosaic karyotypes were found in six patients. One of them had a karyotype 46,XX/46,XX,r(14) and the others had sex chromosomal mosaicisms. The prenatal diagnosis could be essential in these cases. It is concluded that chromosomal aberrations could play a role in the repeated failure of ART procedures. PMID- 10690679 TI - EPH-gestosis (pre-eclampsia)-induced decrease of gelatinase activity may promote an accumulation of collagen in the umbilical cord artery. AB - It was found in our previous paper that edema, proteinuria, hypertension (EPH) gestosis-associated accumulation of collagen in the umbilical cord artery (UCA) is a result of increased biosynthesis and decreased degradation of this protein. It is known that the activity of collagenolytic enzymes is a main factor regulating collagen degradation rate in various tissues. For this reason it was decided to evaluate the effect of EPH-gestosis on the activity of proteolytic enzymes which may be involved in collagen degradation in the UCA wall. Proteolytic activity against bovine serum albumin, reconstituted collagen fibres and gelatin were evaluated. Latent forms of proteolytic enzymes were activated by the action of trypsin, p-chloromercuric benzoate (PCMB) and p-aminophenylmercuric acetate (APMA). A low activity of gelatinase (type IV collagenase) was detected in the extracts from the wall of the umbilical cord artery. This enzyme increased its activity several times after the action of trypsin, PCMB and APMA. EPH gestosis results in a distinct reduction in gelatinase activity. Despite the action of activating agents the gelatinase from EPH-gestosis UCAs was considerably lower in comparison to control UCAs. It can be concluded that gelatinase of the umbilical cord artery forms an inactive complex with a tissue inhibitor of metalloproteinases. Such a complex dissociates under the action of trypsin, PCMB or APMA or sodium dodecyl sulphate. The decrease of gelatinolytic activity in the umbilical cord artery may be a factor that reduces the breakdown of collagen in the arterial wall and promotes an accumulation of this protein. The accumulation of collagen with simultaneous reduction in elastin content in the UCA may be the factors which reduce the elasticity of arterial wall and decrease the blood flow in the fetus of woman with EPH-gestosis. PMID- 10690680 TI - Treatment outcome in women with a single ovary versus patients with two ovaries undergoing in vitro fertilization and embryo transfer (IVF/ET). AB - OBJECTIVE: To compare ovarian response and pregnancy rate between women with one and two ovaries undergoing in vitro fertilization and embryo transfer (IVF/ET). STUDY DESIGN: 20 IVF/ET treatment cycles in ten women with a single ovary were compared with 60 IVF/ET cycles in 47 women with two ovaries. Both groups were matched for age and treated for mechanical infertility. In both groups treatment protocol included gonadotropin releasing hormone/human menopausal gonadotropin/human chorionic gonadotropin (GnRH/hMG/hCG). RESULTS: Effective daily dose of gonadotropins (3.7+/-0.7 vs. 3.6+/-1.0), mean 17beta-estradiol levels on day of hCG administration (1136+/-467 vs. 1343+/-776), number of retrieved oocytes (6.4+/-3.7 vs. 8.3+/-4.2) and number of embryos per transfer (3.0+/-0.7 vs 2.9+/-1.2) were not statistically different between the groups. A significantly higher pregnancy rate was observed among women with one ovary (52.9%) as compared with those with two ovaries (20.8%), (P=0.015). Multivariate logistic regression analysis demonstrated an odds ratio of 5.73 for patients with a single ovary. CONCLUSION: Treatment outcome in patients with a single ovary undergoing IVF/ET is comparable to those with two ovaries. The unexpected significantly higher pregnancy rate observed among these patients need to be further evaluated. PMID- 10690681 TI - The Mirror syndrome. AB - This is a case report illustrating a patient who developed pre-eclampsia with non immunological hydrops fetalis associated with fetal tachycardia. It illustrates how successful treatment of a fetal disorder can lead to resolution of the maternal disorder. PMID- 10690682 TI - A case report of acute pelvic thrombophlebitis missed by magnetic resonance imaging of the pelvic veins. AB - A 29-year-old woman presented post-natally with pulmonary hypertension. Peripheral venous thrombosis was not detected by duplex ultrasound or conventional MRI. Despite anticoagulation, the patient arrested. Autopsy revealed right iliac vein thrombosis. The ability of conventional MRI to detect acute pelvic thrombophlebitis depends on obtaining appropriate views. PMID- 10690683 TI - Perinatal outcome in adolescent pregnancies: a case-control study from a Turkish university hospital. AB - OBJECTIVE: To determine whether adolescent pregnancy is associated with increased risk for adverse pregnancy outcome. STUDY DESIGN: Retrospective case-control study which enrolled 1460 singleton adolescent pregnancies and 2980 controls, delivered at Hacettepe University Hospital between January 1990 and January 1998. RESULTS: Significantly higher rate of perinatal and severe perinatal complications were noted in adolescents. The presence of historical risks, multiparity, young age and lack of prenatal care were significant predictors of these complications. Exclusion of them except for age, revealed comparable perinatal and severe perinatal complications in both groups. CONCLUSION: Adolescent women who receive adequate prenatal care are at no greater risk of an adverse obstetric outcome than adult women of a similar sociodemographic background. PMID- 10690684 TI - Borderline form of empty follicle syndrome: is it really an entity? AB - We report two patients who were administered erroneously 1000 IU HCG instead of 10,000 IU for ovulation triggering in an assisted reproductive technology (ART) cycle. In one case no oocyte was retrieved from eight mature follicles after several washing attempts and retrieval day serum beta-HCG level was 21 mIU/ml. As there was no follicle in the other ovary we did not try a rescue protocol. In the other case two immature and one mature oocytes were retrieved from 15 mature follicles located in both ovaries. Retrieval day serum beta-HCG level was 12 mIU/ml. ICSI was performed in one mature and two in vitro maturated oocytes and the embryo transfer that was done 72 h after the retrieval yielded a healthy singleton pregnancy. Our experience shows that a borderline form of EFS may be an entity and it may be possible to see the formation of periovulatory events and even to obtain a pregnancy in an ART cycle where the ovulation was triggered by 1000 IU HCG. PMID- 10690685 TI - Epileptogenic activity of folic acid. PMID- 10690686 TI - Anticoagulation of older patients. PMID- 10690687 TI - Adverse drug reactions. PMID- 10690688 TI - What's the use of cranberry juice? PMID- 10690689 TI - Pacemaker syndrome in older people. PMID- 10690690 TI - Beta-blocker use in elderly patients with coronary heart disease. PMID- 10690691 TI - Non-drug strategies to resolve psycho-social difficulties after stroke. PMID- 10690692 TI - A comparison of a low-dose warfarin induction regimen with the modified Fennerty regimen in elderly inpatients. AB - OBJECTIVES: To compare a new low-dose warfarin induction regimen with the Fennerty regimen in elderly inpatients. DESIGN: Age-stratified, randomized prospective study. SUBJECTS: 120 age-stratified elderly inpatients. INTERVENTIONS: Each patient was randomized to either the new induction regimen or to a modified Fennerty regimen. MAIN OUTCOMES MEASURES: Days to therapeutic International Normalized Ratio (INR >2); days in the therapeutic range (INR 2-3) during induction; number of patients with INR >4.5; ability of day 4 INR to predict day 8 warfarin dose. RESULTS: The mean time to therapeutic INR was longer for the new induction regimen than modified Fennerty regimen in patients aged 65 75 years [4.6 (mean) +/- 1.6 (SD) days vs 3.8 +/- 0.8 days; P = 0.03] and in patients aged >75 years (4.5 +/- 1.4 days vs 3.5 +/- 0.7 days; P = 0.003). Patients spent more time in the therapeutic INR range with the new induction regimen [3.0 +/- 1.3 days vs 2.7 +/- 1.3 days (P = 0.03) for those aged 65-75 years and 2.9 +/- 1.1 days vs 2.4 +/- 1.3 days (P = 0.04 for those aged >75 years]. Fewer patients using the new regimen had INRs >4.5 in the first 8 days [1 (3%) vs 6 (20%) for 65-75 years (P < 0.05) and 1 (3%) vs 11 (37%) for >75 years (P < 0.01)]. The ability to predict the maintenance dose to within 1 mg was 55% for both regimens. CONCLUSION: The low-dose regimen has important clinical advantages over the Fennerty regimen for anticoagulating elderly inpatients. PMID- 10690693 TI - Contribution of adverse drug reactions to hospital admission of older patients. AB - OBJECTIVE: To describe the severity of adverse drug reactions as a factor in hospital admission of older patients, and to identify risk indicators for severe adverse drug reactions in these patients. DESIGN: Observational cross-sectional study. SETTING: Five wards in a university hospital in The Netherlands. SUBJECTS: Patients aged 70 and over admitted to general medical wards. METHODS: Use of statistical comparison and Kramer's algorithm. RESULTS: A severe adverse drug reaction was present in 25 (24%) of 106 patients. Thirteen patients (12%; 95% confidence interval 6.1-18.6%) were admitted probably because of an adverse drug reaction. Risk indicators for a severe adverse drug reaction were a fall before admission (odds ratio 51.3, P = 0.006), gastrointestinal bleeding or haematuria (odds ratio 19.8, P < 0.001) and the use of three or more drugs (odds ratio 9.8, P = 0.04). CONCLUSION: Adverse drug reactions are an important cause of hospital admissions in older people. A fall before admission may indicate a severe adverse drug reaction. PMID- 10690694 TI - Measuring symptom change in patients with Parkinson's disease. AB - BACKGROUND: The 39-item Parkinson's disease questionnaire (PDQ-39) is more sensitive to functional change than other measures of health and disability. AIM: To determine the ability of this scale to change over time and the concurrent validity of some of its subscales. METHODS: We assessed a cohort of 67 Parkinson's patients for 18 months, using the PDQ-39, the GHQ-28 general health questionnaire and the Office of Population and Census Surveys disability instrument. RESULTS: The Office of Population and Census Surveys disability instrument and GHQ-28 recorded no significant change, but the PDQ-39 showed marked changes in levels of functioning. We also analysed changes on the PDQ-39 subscales as well as concurrent validity data for several subscales. This showed concurrent validity with the Beck depression and anxiety inventories, the Barthel index and the Royal Postgraduate Medical School severity scale. There was a high level of concurrent validity for all comparisons except for the Barthel index. CONCLUSION: The PDQ-39 is a sensitive tool for monitoring change in patients with Parkinson's disease. It has high levels of concurrent validity with established measures of mood and motor function. PMID- 10690695 TI - The association between atrophic glossitis and protein-calorie malnutrition in old age. AB - AIM: To examine the relationship between atrophic glossitis (absence of papillae in more than 50% of the tongue) and nutritional status. DESIGN: A randomized population survey. SETTING: The medical department of Aker University Hospital, and people living at home. PARTICIPANTS: 310 old people recently admitted to hospital and 106 randomly selected elderly people at home. MEASURES: The presence of atrophic glossitis and nutritional indices. RESULTS: Atrophic glossitis occurred in 13.2% of men and 5.6% of women at home and in 26.6% of men and 37% of women in hospital. The atrophic glossitis was related to reduced weight, body mass index, triceps skinfold thickness, arm-muscle circumference, muscular strength, activities of daily living and serum concentrations of cholesterol, ascorbic acid, cholecalcidiol and B12, but not to levels of zinc or folate. In a multiple logistic regression model, atrophic glossitis was related only to cholesterol (P = 0.032), muscular strength (P = 0.018) and activities of daily living (P = 0.03). CONCLUSION: Atrophic glossitis is common in elderly people and is a marker for malnutrition and reduced muscle function. PMID- 10690696 TI - Protein-energy oral supplementation in malnourished nursing-home residents. A controlled trial. AB - OBJECTIVES: To validate a nutritional intervention programme for elderly people living in nursing homes. DESIGN: In a prospective, randomized, controlled study of 88 residents, we determined nutritional status at day 0 and day 60 using a record of dietary intake, anthropometry, hand-grip strength and mini-nutritional assessment. Dietary intake, grip strength and body weight were also recorded at day 30. We divided subjects into four groups according to their mini-nutritional assessment score. Those with a score 24 received no oral supplementation. Those at risk of malnutrition (with a score of 17-23.5) were randomized to oral supplementation. Those with a score <17 received oral supplementation. We recorded the amount of oral supplements consumed daily. RESULTS: Compliance with oral supplementation was good, and daily intake averaged about 400 kcal. The total energy intake on day 60 was significantly higher in both of the groups that received supplements. Following supplementation, most subjects at risk of malnutrition improved their mini-nutritional assessment score and increased their weight (by 1.4 +/- 0.5 kg). Neither the mini-nutritional assessment score nor weight improved in subjects at risk of malnutrition who did not receive supplements. Supplementation in the malnourished group resulted in a mean mini nutritional assessment score increase (from 13.9 +/- 2.6 to 17.1 +/- 3.9) and a mean weight gain of 1.5 +/- 0.4 kg. CONCLUSION: Oral nutritional supplements are well accepted and result in increased daily protein and energy intake, body weight and nutritional status in most malnourished patients and in those at risk of malnutrition. PMID- 10690697 TI - Hip protectors improve falls self-efficacy. AB - OBJECTIVES: To investigate the effect of use of external hip protectors on subjects' fear of falling and falls self-efficacy (belief in their own ability to avoid falling). DESIGN: Randomized controlled trial. SETTING: Aged-care health services in Sydney, Australia. PARTICIPANTS: 131 women aged 75 years or older, who had two or more falls or one fall requiring hospital admission in the previous year and who live at home. Sixty-one subjects were in the intervention group and 70 in the control group. INTERVENTION: Use of external hip protectors and encouragement to use the protectors by an adherence nurse. MEASUREMENTS: At the time of enrolment into a wider study examining the effect of hip protectors on hip fractures, participants recruited at home completed an assessment of fear of falling and falls efficacy as measured by the Falls Efficacy Scale and the Modified Falls Efficacy Scale. At 4-month follow-up, these scales were readministered by an observer who was not aware of the allocation of the participant to intervention or control groups. RESULTS: Fear of falling and falls self-efficacy, as measured by the Falls Efficacy and Modified Falls Efficacy Scales, were similar at baseline in both groups. Fear of falling was present at follow-up in 43% of subjects using hip protectors and 57% of the control group (chi2 = 2.58, P = 0.11). Hip protector users had greater improvement in falls self-efficacy at follow-up as measured by the Falls Efficacy Scale (t = 2.44, P = 0.016) and the Modified Falls Efficacy Scale (t = 2.08, P = 0.039). CONCLUSION: Hip protectors improve falls self-efficacy. As users of hip protectors feel more confident that they can complete tasks safely, they may become more physically active and require less assistance with activities of daily living. PMID- 10690698 TI - The Dutch pressure sore assessment score or the Norton scale for identifying at risk nursing home patients? AB - OBJECTIVE: To investigate the usefulness of a Dutch pressure sore risk assessment scale--the Centraal Begeleidingsorgaan voor de Intercollegiale Toetsing (CBO; National Organization for Quality Assurance in Hospitals) score--in the detection of patients at risk of developing pressure sores after admission to a nursing home. As the Norton score is the standard method of risk assessment, we also investigated which score (Norton or CBO) has the stronger relationship to the development of pressure sores. DESIGN: Longitudinal cohort design. PATIENTS: 220 nursing home patients, 80 men, 140 women, mean age 79 years (standard deviation 3). MEASURES: Admission assessments for the presence of pressure sores, CBO and Norton scores, preventive measures and demographic characteristics. We made observations every week for 4 weeks. MAIN OUTCOME MEASURE: Presence or absence of pressure sores. MAIN RESULTS: 54 patients (25%) developed a pressure sore. A significant, nonlinear relationship was found between the CBO score on admission and the development of pressure ulcers for the first 2 weeks after admission. Multiple logistic regression analysis showed that only mobility (odds ratio = 3.6, P = 0.0001) and mental state (odds ratio = 2.0, P = 0.03) showed a significant relationship with the development of pressure ulcers. The CBO score was no better in risk assessment than the Norton score. CONCLUSIONS: The CBO score can be used for assessment of the risk of developing pressure ulcers in the first 2 weeks after admission to a nursing home, but is no better than the Norton score. Since the Norton score is easier to use, it is slightly preferable for use in this setting. However, neither score is a good indicator of patients at risk. Physicians should not depend solely on risk scores when prescribing preventive measures. PMID- 10690699 TI - Serum cholesterol concentrations and all-cause mortality in older people. AB - OBJECTIVE: To study the impact of serum cholesterol concentrations on the total risk of mortality in older people. DESIGN: Retrospective cohort study with a follow-up of 8-10 years. SUBJECTS: A total of 989 subjects (367 men and 622 women) aged 65 and over, living in the Marshfield Epidemiologic Study Area at the time of their first complete serum lipid assessment. METHODS: We calculated sex specific mean levels of serum total cholesterol, low-density lipoprotein, high density lipoprotein and triglycerides, and the ratio of total cholesterol to high density lipoprotein, for subjects who died of all causes and for those who survived to the end of follow-up, with adjustment for relevant covariates. We obtained estimates of the risk factor-adjusted sex-specific relative risk for all cause mortality with approximate quartiles of serum cholesterol concentrations by proportional hazards regression models. We also evaluated the possible combined effects of age, sex and cholesterol on all-cause mortality. RESULTS: A high level of high-density lipoprotein was significantly associated with a low total risk of mortality in older men. Conversely, an elevated ratio of total cholesterol to high-density lipoprotein was directly related to an increased total risk of mortality in older men. Age and high-density lipoprotein level had a significant synergistic effect on all-cause mortality for the elderly men. We found little or no association in women between all-cause mortality and any of the lipid measures studied. CONCLUSIONS: An increased ratio of total cholesterol to high-density lipoprotein appears to be associated with an increase in risk for all-cause mortality in men aged 65 and over, while an elevated level of high-density lipoprotein, considered alone, seems to be protective against mortality from all causes in men aged 65-74 years, but this effect diminishes over the age of 75. PMID- 10690700 TI - Reproductive longevity and increased life expectancy. AB - BACKGROUND: Female life expectancy in developed countries has increased by 30 years in the twentieth century. AIM: To determine if there has been an increase in reproductive longevity. METHODS: We analysed age-specific fertility data from birth statistics for the USA, Canada, Japan, France, Sweden, the UK and Australia. RESULTS: Since 1940, birth rates for women aged 35 and over have declined. Among women aged 50 years and older, there has been no increase in births. Fertility rates in 1990 were 0.0 to 0.044 per 1000 women, with total numbers ranging from 0 to 60 births. CONCLUSION: The fertile years have not been prolonged in the cohort of women whose life expectancy has increased so dramatically this century. This suggests that reproductive senescence is tightly controlled and not extended by factors that enhance female longevity. Other physiological mechanisms may also be fixed within narrow age limits. PMID- 10690701 TI - Do older hospital patients recognize adverse drug reactions? AB - OBJECTIVE: To establish the relationship between subjective complaints of side effects of drugs and the objective presence of adverse drug reactions in older patients. DESIGN: Observational cross-sectional study. SETTING: Five medical wards at the University Hospital Rotterdam Dijkzigt. SUBJECTS: Patients aged 70 and over admitted to the general medical wards over a 3-month period. METHODS: Statistical comparison and Kramer's algorithm. RESULTS: Of 106 patients, 102 used medication, and 93 of these were able to report whether they believed they were experiencing drug side effects. Thirty-six [39% (95% confidence interval 28.8 48.6)] believed that they were experiencing side effects and the number of diagnoses per patient and the proportion of patients with chronic obstructive pulmonary disease was higher in these 36 'complainers' than in the group of the 'non-complainers'. We found a correct opinion (true positive and negative) about the objective presence or absence of mild or severe adverse drug reactions in 79% (95% confidence interval 70.2-86.8). Asking the patient about side effects of drugs had a sensitivity of 0.70 and a specificity of 0.85 patients. The severe adverse drug reactions in 21 patients were not recognized by 14 of them. CONCLUSION: At hospital admission, older patients should be asked about drug side effects because they are often correct in recognizing them. However, severe adverse drug reactions are not easily recognized. PMID- 10690702 TI - Myasthenia gravis and recurrent falls in an elderly patient. AB - PRESENTATION: An elderly man had recurrent hospital admissions with falls. OUTCOME: Acetylcholine receptor antibodies and single-fibre electromyogram were useful in the diagnosis of myasthenia gravis. Treatment prevented further hospital admissions. PMID- 10690703 TI - Carotid sinus hypersensitivity--a modifiable risk factor for fractured neck of femur. PMID- 10690704 TI - Proximal femoral fracture while wearing correctly applied hip protectors. PMID- 10690705 TI - Why do so few older people with aortic stenosis have valve replacement therapy? PMID- 10690706 TI - Evaluation of an integrated care pathway for stroke unit rehabilitation. PMID- 10690707 TI - Fetal biometry in different ethnic groups. AB - OBJECTIVE: The aim of this study was to determine whether differences in ultrasound-measured fetal biometry exist between pregnant woman of autochthonous Belgian origin and migrant women from Morocco and Turkey. METHOD: A prospective cross-sectional study was performed in which fetal biparietal diameter, head circumference, abdominal circumference and femur length were measured in pregnant women presenting between 18 and 40 weeks of gestation. Fetal weight was calculated using the formulae by Shepard and Hadlock. Only uncomplicated singleton pregnancies with a certain date of the last menstrual period, confirmed by early ultrasound, were included. The father of the child had to be of the same ethnic origin as the mother. Polynomial regression of the different measurements was performed for women of autochthonous Belgian origin and for migrant women from Morocco and from Turkey. RESULTS: Singleton fetuses numbering 524 were examined, including 369 Belgian, 78 Moroccan and 77 Turkish. Polynomial regression was performed for the three groups for the biparietal diameter, head circumference, abdominal circumference, femur length and estimated fetal weight. No significant difference between the three different ethnic groups could be demonstrated for the biparietal diameter (P = 0.39). There was a significant difference for the head circumference (P = 0.017), the abdominal circumference (P = 0.0015), the femur length (P = 0.0014) and the estimated fetal weight for both formulae (Shepard P = 0.047; Hadlock P = 0.0006). CONCLUSION: In this set of cross-sectional data no significant difference for ultrasound-measured fetal biparietal diameter between autochthonous Belgian women and migrant women from Morocco and from Turkey could be demonstrated. Differences do exist for the head circumference, the abdominal circumference, the femur length and the estimated fetal weight. The use of adapted charts of fetal size for pregnant women of Turkish or Moroccan origin should be considered. PMID- 10690708 TI - Developmental changes of vitamin K epoxidase and reductase activities involved in the vitamin K cycle in human liver. AB - We examined the developmental changes in activities of vitamin K epoxidase, and vitamin K-2,3-epoxide reductase and vitamin K reductase in the human autopsied liver. The activity of epoxidase, which converts vitamin K hydroquinone to its epoxide, showed a high value in the early prenatal period of 10-30 gestational weeks but decreased rather rapidly in contrast with the reductase activities. After birth, a significant decrease of the epoxidase activity was observed but no such change was seen during the postnatal period. On the other hand, the activities of vitamin K-2,3-epoxide reductase and vitamin K reductase, which convert vitamin K-2,3-epoxide to its hydroquinone, showed a significantly low value in the early prenatal period. The highest activity of vitamin K epoxidase in the early prenatal period may be essential to the production of vitamin K dependent ligands for growth factors expressed in the embryo. PMID- 10690709 TI - Variation of the ganglioside compositions of human milk, cow's milk and infant formulas. AB - The ganglioside compositions of human milk, cow's milk and infant formulas were compared. The results showed that there was a drastic change in the ganglioside composition from the colostrum to later human milk, and that both the patterns and contents of gangliosides in human milk, cow's milk and infant formulas differed markedly. In human milk, the total lipid-bound sialic acid level was two times higher than those in cow's milk and infant formulas. The major ganglioside in the later human milk, GM3 (27.7%), was only a minor component in the colostrum, cow's milk and infant formulas (3.3, 2.8 and 0.4-2.6%, respectively). GD3 represented 49.0, 61.0 and 72.4-86.6%, respectively, of the colostrum, cow's milk and infant formulas, compared to 31.8% of the later human milk gangliosides. Another four gangliosides, which were assumed to be c-series gangliosides, were detected in the colostrum and the later human milk. They represented 33-38% of total lipid-bound sialic acid, and were tentatively designated as GX1, GX2, GX3 and GX4, respectively. However, only GX1 and GX2 were observed in cow's milk and infant formulas. The variation of the gangliosides in human and cow's milk, and infant formulas might have some biological significance regarding neonatal brain development, allergies, infant growth and non-immunoglobulin prophylactic activities against some bacterial toxins. PMID- 10690710 TI - Visual-motor function of very low birth weight and full-term children at 3 1/2 to 4 years of age. AB - Improvements in perinatal and neonatal management have not only led to a higher survival rate of very low birth weight infants (VLBW; < or = 1,500 g or < 32 weeks gestational age), but also to a better outcome of these children. However the percentage of VLBW children who need special education because of later school problems remains high even in children considered neurologically normal during infancy. We assessed 40 VLBW children and 83 healthy full-term children at age 3 to 4 years by means of a simple and short test for visual-motor deficits. The test included the copying and cutting-out of geometric shapes, the building of models, the recognition of colours and the observation of the concentration and cooperation during the test. All VLBW children had had a good perinatal outcome and had been considered neurologically normal at one year of age. Most VLBW children scored within 1 standard deviation (S.D.) of the test mean, but on average the VLBW children scored significantly lower than the full-term infants in the copying of figures, the cutting-out of geometric forms, the building of models and in the overall concentration and cooperation during the test. Children who attended a nursery school achieved significantly better test results. Girls tended to have better results, but this was not statistically significant. Social factors and age had a significantly greater impact on results than perinatal factors. In summary, VLBW children scored significantly less in almost every test item compared to their term peers. Our test battery could serve as a short introductory test to screen for deficits in visual-motor skills, especially in VLBW children. PMID- 10690711 TI - Efficiency of left ventricular diastolic function increases in healthy full-term infants during the first months of life. A prospective follow-up study. AB - Postnatal changes in left ventricular diastolic filling and systolic cardiac performance were studied monthly by serial echocardiographic measurements from days 3-5 up to six months in 20 healthy full-term infants. The fractional shortening area (FSA = (left ventricular end-diastolic area - end-systolic area)/end-diastolic area) was assessed for systolic performance, and transmitral pulsed-wave Doppler flow velocity patterns were analysed for diastolic function. FSA remained stationary during the observation. After birth, left ventricular peak early (E) and atrial (A) velocities and the respective integrals were lower than at one month of age (47+/-5 vs. 63+/-6 cm/s and 44+/-6 vs. 57+/-5 cm/s and 3.33+/-0.40 vs. 4.05+/-0.53 cm and 2.74+/-0.40 vs. 3.18+/-0.53 cm; P < 0.05). During the next five months, the early parameters (E velocity and E integral) increased but the atrial indices (A velocity and A integral) did not change. During the whole observation the E/A velocity ratio, the E/A integral ratio and the early filling fraction (EFF) increased. The early filling deceleration time was longer during the first month than later (87+/-10 vs. 72+/-13 ms; P < 0.05). In conclusion, age-related changes were observed in the diastolic but not in the systolic performance in healthy full-term infants during the first six months. The most intensive changes took place in the early and atrial transmitral parameters during the first month of life, suggesting an improvement in both left ventricular relaxation and compliance. During the following five months, the early mitral parameters increased but the atrial diastolic values remained stable. These changes may mainly be determined by the improvement in left ventricular relaxation. PMID- 10690712 TI - Effect of epidermal growth factor on lung growth in experimental fetal pulmonary hypoplasia. AB - The purpose of this study was (1) to compare the expression of epithelial growth factor receptor (EGFR) in the lung tissues of human fetuses with or without pulmonary hypoplasia, and (2) to investigate the effects of EGF on lung growth in experimental pulmonary hypoplasia in rabbits. Firstly, we investigated the expression of EGFR in lung tissues of human fetuses with or without pulmonary hypoplasia by immunohistochemistry. Secondly, the amniotic fluid was shunted into the maternal abdominal cavity in a group of 12 fetal rabbits, another group (n = 12) received EGF injection (5 microg, i.p.) at day 25 of gestation. The third group (n = 12) was only treated with EGF while littermates not operated on served as the control group (n = 12). On day 29 of gestation, fetuses were delivered by Cesarean section and the lungs removed. The body weight and wet lung and liver weights were measured. As a measure of fetal lung growth, we determined the size of lung acini, the number of terminal airspaces, and diameter of alveoli (n = 6, each groups). We also measured the concentration of phosphatidylcholine (PC) and the lecithin/sphingomyelin (L/S) ratio in lung lavage fluid at birth in some fetuses (n = 6, each groups). In human fetuses with pulmonary hypoplasia, there was a significant decrease in radial alveolar count and expression of EGFR compared with fetuses without pulmonary hypoplasia. Amniotic shunt significantly decreased fetal lung/body weight ratio compared with control. Injection of EGF in the shunted group significantly increased lung/body weight ratio to the control level. The concentration of PC and L/S ratio in lung fluid lavage from rabbit fetuses with hypoplastic lungs was significantly higher than the other three groups. Histopathological examination of fetuses with hypoplastic lungs treated with EGF showed no significant change in the size of acini, number of terminal airspaces or the diameter of alveoli compared with the control group. Our results suggested that EGF was associated with lung growth and maturation of human lung and that treatment of rabbit fetuses with hypoplastic lungs with EGF facilitated lung growth and development. PMID- 10690713 TI - Hepatic taurine, glycine and individual bile acids in early human fetus. AB - OBJECTIVE: To examine 3alpha-hydroxy individual bile acid and the association between taurine and taurine-conjugated bile acid in human fetal liver. METHODS: Taurine, glycine and the individual bile acids in liver samples taken from 17 human fetuses whose abortion was induced from 12 to 23 weeks' gestation were determined by high-performance liquid chromatography. RESULTS: The concentrations of taurine, glycine and bile acids in early fetal life were not related to gestational age. Hepatic taurine was statistically correlated with total taurine conjugates (r = 0.798; P < 0.0001), but hepatic glycine was not correlated with glycine-conjugates (r = -0.103; P > 0.05). Lithocholic acid was conjugated exclusively with taurine. CONCLUSIONS: Our results supported that in human fetal life there is a considerable bile acid pool in liver, and that taurine works effectively on the bile acid metabolism depending on the hepatic taurine concentration. PMID- 10690714 TI - Conquering pain. PMID- 10690715 TI - Microsurgical resection of brainstem, thalamic, and basal ganglia angiographically occult vascular malformations. AB - OBJECTIVE: To evaluate the clinical results for patients who underwent resection of angiographically occult vascular malformations (AOVMs) of the brainstem, thalamus, or basal ganglia, successfully resected after it exhibited rebleeding and presented to a pial surface. METHODS: Between January 1990 and May 1998, 56 patients with 57 deep AOVMs underwent 63 operations, at Stanford University Medical Center, to treat AOVMs of the brainstem (42 AOVMs), thalamus (5 AOVMs), or basal ganglia (10 AOVMs). The surgical approach was suboccipital midline (27 operations), far lateral suboccipital (10 operations), transsylvian (9 operations), interhemispheric transcallosal or infracallosal (8 operations), infratentorial supracerebellar (6 operations), or subtemporal (3 operations). Four patients experienced recurrent bleeding from the same lesion after surgical resection, requiring a second operation. One patient required a planned second operation, using a different approach, to completely resect the lesion, and one patient underwent two surgical procedures to resect two separate brainstem AOVMs. One patient initially underwent exploration but not resection of her AOVM, because it did not present to a pial or ependymal surface. The AOVM was successfully resected after it exhibited rebleeding and presented to a pial surface. RESULTS: The immediate outcomes after surgery were unchanged for 31 patients (55%), worsened for 16 (29%), and improved for 9 (16%). The long-term outcomes were unchanged for 24 patients (43%), compared with their presenting grade, worse for 3 (5%), and improved for 29 (52%). Patients who had undergone previous radiotherapy or radiosurgery to treat these lesions experienced more difficult postoperative courses, and radiation necrosis was observed for two patients. CONCLUSION: AOVMs of the brainstem, thalamus, and basal ganglia can be safely removed, with a long-term neurological morbidity rate of only 5% and a complete lesion resection rate of 93% after the initial planned resection. The use of cranial base surgical approaches and intraoperative electrophysiological monitoring contributes to successful clinical outcomes. PMID- 10690716 TI - Factors associated with intracranial hemorrhage in cases of cerebral arteriovenous malformation. AB - OBJECTIVE: The standard categorization of arteriovenous malformations (AVMs) involves the Spetzler-Martin grading system, which uses a simple analysis of size, location (superficial or deep), and the presence of deep or superficial drainage. Hemodynamic risk factors are also thought to play important roles in the pathogenesis of these lesions and to be associated with the intracranial hemorrhage (ICH) rate. The actual hemodynamic factors for AVMs cannot be easily measured, but angioarchitectural features can be assessed and used as surrogate parameters. METHODS: The AVM angioarchitectural features for 662 patients were analyzed, and their associations with ICH as a presenting sign were studied. A cross-sectional analysis was used to qualify the strength of associations among clinical features, angioarchitectural characteristics, and ICH before treatment. RESULTS: The multivariate analysis indicated that arterial stenosis and arterial ectasia were associated with lower ICH rates, whereas venous stenosis increased the rate of ICH. The presence of angiogenesis modified the effects of arterial and venous stenosis. Furthermore, the effect of venous stenosis depended on the location of the nidus. The presented data do not support a direct positive association between associated aneurysms and ICH. CONCLUSION: Certain angiographic features seem to have prognostic potential with respect to the occurrence of ICH among patients with AVMs. A discriminatory prognostic index is proposed; its relevance must be proven in a future prospective study. PMID- 10690717 TI - Ischemic events associated with unruptured intracranial aneurysms: multicenter clinical study and review of the literature. AB - OBJECTIVE: To determine the prevalence, clinical characteristics, and long-term outcomes in cases involving transient ischemic attacks (TIAs) or ischemic strokes secondary to embolization from unruptured intracranial aneurysms. METHODS: We identified all available patients with intracranial aneurysms and ischemic strokes in three university-affiliated hospitals, using either International Classification of Diseases-9th Revision codes or local registries. Patients with clinically or radiologically detected cerebral infarctions distal to intracranial aneurysms, in the absence of other causes for the infarctions, were included. An aneurysmal embolic source was considered highly probable by the primary neurosurgeon/neurologist in all cases. Follow-up data for the patients were acquired through reviews of clinical visits or telephone interviews. A review of the literature was performed to identify characteristics of previously reported patients. RESULTS: Ischemic strokes or TIAs attributable to embolization from the aneurysmal sac were observed for 9 of 269 patients (3.3%) with unruptured aneurysms. Of these nine patients, five were women and four were men (mean age, 62 yr; age range, 45-72 yr). Symptomatic aneurysms were located in the middle cerebral artery (n = 4), internal carotid artery (n = 3), posterior cerebral artery (n = 1), or vertebral artery (n = 1). The mean maximal diameter was 12.5 mm (range, 5-45 mm). Six patients underwent surgical treatment, of whom two experienced postoperative cerebral infarctions referable to the distribution of the artery harboring the aneurysm. Two patients were treated with aspirin, and one patient received no treatment. The mean follow-up period was 38 months (range, 1-60 mo). None of the patients experienced additional ischemic events during the follow-up period. Among the 41 previously reported patients, conservative treatment was used for 20 patients (mean follow-up period, 50.7 +/- 44.5 mo). Four of the 20 patients experienced recurrent TIAs, 1 patient experienced worsening of symptoms, and 1 patient died during the follow-up period. A total of 21 patients underwent surgical treatment (mean follow-up period, 33.6 +/- 32.3 mo). Of these patients, only one experienced recurrent TIAs. Two patients experienced postoperative seizures, and one patient died during the follow-up period. All recurrent symptoms with either surgical or conservative treatment were transient, and no patient experienced a major or disabling stroke during the follow-up period. CONCLUSION: Ischemic events can occur distal to both small and large unruptured intracranial aneurysms (predominantly in the anterior circulation). The long-term risk of recurrent ischemic events, particularly major or disabling strokes, seems to be low with either surgical or conservative treatment. PMID- 10690718 TI - Long-term outcomes for surgically resected craniopharyngiomas. AB - OBJECTIVE: This retrospective study critically analyzed the long-term functional outcomes and tumor recurrence rates for surgically treated craniopharyngiomas. METHODS: This study used an outcome classification system that included functioning vision, independent versus dependent living, Karnofsky Performance Scale scores, academic levels, work status, and psychological status. Tumor recurrence rates were analyzed with respect to the extent of surgical resection and adjunctive radiotherapy. RESULTS: For 121 patients, with a mean follow-up period of 10 years, the overall "good outcome" rate was 60.3%. Factors associated with poor outcomes included lethargy at presentation, visual deterioration, papilledema, tumor calcification, hydrocephalus, and tumor adhesiveness at surgery. Gross total resection was associated with good outcomes (P = 0.017) and decreased risk of recurrence (P = 0.024). Subtotal resection was associated with increased risk of tumor recurrence (P = 0.0235). The highest risk of recurrence was in the subtotal resection/no radiation group (P = 0.0001). There were no differences in outcomes or recurrence rates between pediatric and adult patients. There were also no differences in outcomes or recurrence rates between papillary and adamantinous tumors. Approximately one-third of patients exhibited morbid obesity, and permanent diabetes insipidus was observed for 25 patients. CONCLUSION: A rigorous evaluation of outcomes for tumors such as craniopharyngiomas must consider not only the extent of resection, as judged by postoperative imaging, but also the long-term physical, intellectual, and psychological functioning of the patients. PMID- 10690719 TI - Using proton magnetic resonance spectroscopic imaging to predict in vivo the response of recurrent malignant gliomas to tamoxifen chemotherapy. AB - OBJECTIVE: Most patients with a malignant glioma spend considerable time on a treatment protocol before their response (or nonresponse) to the therapy can be determined. Because survival time in the absence of effective therapy is short, the ability to predict the potential chemosensitivity of individual brain tumors noninvasively would represent a significant advance in chemotherapy planning. METHODS: Using proton magnetic resonance spectroscopic imaging (1H MRSI), we studied 16 patients with a recurrent malignant glioma before and during treatment with high-dose orally administered tamoxifen. We evaluated whether 1H MRSI data could predict eventual therapeutic response to tamoxifen at the pretreatment and early treatment stages. RESULTS: Seven patients responded to tamoxifen therapy (three with glioblastomas multiforme; four with anaplastic astrocytomas), and nine did not (six with glioblastomas multiforme; three with anaplastic astrocytomas). Responders and nonresponders exhibited no differences in their age, sex, tumor type, mean tumor volume, mean Karnofsky scale score, mean number of weeks postradiotherapy, or mean amount of prior radiation exposure. Resonance profiles across the five metabolites measured on 1H MRSI spectra (choline containing compounds, creatine and phosphocreatine, N-acetyl groups, lactate, and lipids) differed significantly between these two groups before and during treatment. Furthermore, linear discriminant analyses based on patients' in vivo biochemical information accurately predicted individual response to tamoxifen both before and at very early treatment stages (2 and 4 wk). Similar analyses based on patient sex, age, Karnofsky scale score, tumor type, and tumor volume could not reliably predict the response to tamoxifen treatment at the same time periods. CONCLUSION: It is possible to accurately predict the response of a tumor to tamoxifen on the basis of noninvasively acquired in vivo biochemical information. 1H MRSI has potential as a prognostic tool in the pharmacological treatment of recurrent malignant gliomas. PMID- 10690720 TI - A preliminary study of the prognostic value of proton magnetic resonance spectroscopic imaging in gamma knife radiosurgery of recurrent malignant gliomas. AB - OBJECTIVE: The goal of this study was to investigate the use of proton magnetic resonance spectroscopic imaging as a prognostic indicator in gamma knife radiosurgery of recurrent gliomas. METHODS: Thirty-six patients with recurrent gliomas were studied with proton magnetic resonance spectroscopic imaging at the time of radiosurgery, and with conventional magnetic resonance imaging examinations at regular time intervals until the initiation of a new treatment strategy. Patients were categorized on the basis of their initial spectroscopic results, and their performance was assessed in terms of change in contrast enhancing volume, time to further treatment, and survival. RESULTS: The trends in the overall population were toward more extensive increase in the percent contrast-enhancing volume, a decreased time to further treatment, and a reduced survival time for patients with more extensive initial metabolic abnormalities. Statistical analysis of the subpopulation of patients with glioblastoma multiforme found a significant increase in relative contrast-enhancing volume (P < 0.01, Wilcoxon signed-rank test), a decrease in time to further treatment (P < 0.01, log-rank test), and a reduction in survival time (P < 0.01, log-rank test) for patients with regions containing tumor-suggestive spectra outside the gamma knife target, compared with patients exhibiting spectral abnormalities restricted to the gamma knife target. Further studies are needed to establish statistical significance for patients with lower-grade lesions and to confirm the results observed in this study. CONCLUSION: The pretreatment spectroscopic results provided information that was predictive of outcome for this patient pool, both in local control (change in contrast-enhancing volume) and global outcome (time to further treatment and survival). This modality may have an important role in improving the selection, planning, and treatment process for glioma patients. PMID- 10690721 TI - In vivo proton magnetic resonance spectroscopy of central neurocytomas. AB - OBJECTIVE: The authors report on the metabolic features of central neurocytomas observed during in vivo single-voxel proton magnetic resonance spectroscopy. METHODS: Volume-selective single-voxel proton magnetic resonance spectroscopy was performed with a 1.5-T unit using a point-resolved spectroscopy sequence (TR/TE = 2000 ms/135 and 270 ms) to obtain spectra of a single 8-cc voxel. The subjects were five patients in the Department of Neurosurgery of Seoul National University Hospital whose central neurocytomas had been diagnosed histologically. The peak intensities of compounds containing choline (Cho), N-acetylaspartate, creatine/phosphocreatine, and lactate were analyzed. RESULTS: The ratios of Cho to creatine/phosphocreatine and Cho to N-acetylaspartate were significantly higher than ratios in normal brains. A lactate signal was present, and an unidentified signal was also observed at 3.55 ppm, which might have been produced by inositol or glycine. CONCLUSION: A combination of the signal at 3.55 ppm and a prominent Cho peak seems to be a characteristic feature of central neurocytomas. Volume-selective single-voxel proton magnetic resonance spectroscopy could provide additional information to aid in diagnosing this condition. PMID- 10690722 TI - The influence of hyperglycemia on neurological outcome in patients with severe head injury. AB - OBJECTIVE: Traumatic brain injury is associated with a stress response that includes hyperglycemia, which has been shown to worsen neurological outcome during cerebral ischemia and hypoxia. To better examine the relationship between hyperglycemia and outcome after head injury, we studied the clinical course of 267 head-injured patients who were admitted for treatment in the neurosurgical department of Asclepeion Hospital of Athens between January 1993 and November 1997. METHODS: We prospectively studied 267 patients with moderate or severe craniocerebral injury (Glasgow Coma Scale scores, 3-12) who were treated surgically for evacuation of an intracranial hematoma and/or placement of a device for intracranial pressure monitoring under general anesthesia to determine the relationship between serum glucose levels, severity of injury, and neurological outcome. RESULTS: Patients with severe head injury had significantly higher serum glucose levels than did those with moderate injury. Patients who subsequently had an unfavorable outcome had significantly higher glucose levels than did those with a better prognosis. Among the patients with more severe head injury, a glucose level greater than 200 mg/dl was associated with a worse outcome. In the same group of patients, a significant relationship was found between postoperative glucose levels, pupillary reaction, and maximum intracranial pressure during the first 24 hours. Multivariate analysis showed that postoperative glucose levels were an independent predictor of outcome. CONCLUSION: Early hyperglycemia is a frequent component of the stress response to head injury, a significant indicator of its severity, and a reliable predictor of outcome. PMID- 10690723 TI - Outcome of unilateral and bilateral pallidotomy for Parkinson's disease: patient assessment. AB - OBJECTIVE: Pallidotomy has recently regained acceptance as a safe and effective treatment for Parkinson's disease symptoms. The goal of this study was to obtain the patients' perspective on their results after undergoing this procedure. Special attention was focused on the potential complications and the respective advantages and risks of unilateral versus bilateral pallidotomy. METHODS: Fifty six patients were studied during a 2-year period; 44 completed the evaluation, with a median follow-up of 7 months. Of these patients, 22 underwent unilateral pallidotomy, and 17 had bilateral simultaneous pallidotomy. Five patients who underwent staged bilateral pallidotomy were excluded from the statistical analysis, because the number of patients was considered too small for analysis. The procedures were performed with magnetic resonance imaging determination of the target, combined with physiological confirmation, including microelectrode recording. RESULTS: According to Visual Analog Scale scores, unilateral pallidotomy significantly improved dyskinesias (P < 0.05) but no other symptoms. Simultaneous bilateral pallidotomy improved slowness, rigidity, tremor, and dyskinesias (P < 0.05) but worsened speech function (P < 0.05). According to the patients' most frequently chosen answers to multiple-choice questions, unilateral pallidotomy improved night sleep, muscle pain, freezing, overall "on," overall "off," and the duration of "off periods," but it worsened the volume of the voice and articulation, increased drooling, and reduced concentration. Bilateral pallidotomy improved night sleep, muscle pain, freezing, overall "on," overall "off," duration of "off periods," and the amount of medication taken, but it increased drooling and worsened the volume of the voice, articulation, and writing. Subjective visual disturbance was noted in 36 and 41% of patients who underwent unilateral and simultaneous bilateral pallidotomy, respectively. Globally, the result of the procedure was rated "good" or "excellent" by 64% of the patients who underwent unilateral pallidotomy and by 76% of the patients who underwent bilateral pallidotomy. An age less than 70 years was a positive prognostic factor for the global outcome (P < 0.05), as were severe preoperative dyskinesias (P < 0.05). CONCLUSION: This study confirms that, from a patient standpoint, unilateral and simultaneous bilateral pallidotomy can reduce all the key symptoms of Parkinson's disease (i.e., akinesia, tremor, and rigidity) and the side effects of L-dopa treatment (i.e., dyskinesias). Preoperative severe dyskinesias and younger age are positive prognostic factors for a successful outcome. Simultaneous bilateral pallidotomy was more effective than unilateral pallidotomy regarding tremor, rigidity, and dyskinesias, but it conferred a higher risk of postoperative speech deterioration. PMID- 10690724 TI - Recurrent trigeminal neuralgia attributable to veins after microvascular decompression. AB - OBJECTIVE: To demonstrate the cause of and optimal treatment for recurrent trigeminal neuralgia (TN) in cases where veins were observed to be the offending vessels during the initial microvascular decompression (MVD) procedure. METHODS: An electronic search of patient records from 1988 to 1998 revealed that 393 patients were treated with MVD for TN caused by veins. The pain recurred in 122 patients (31.0%). Thirty-two (26.2%) of these patients underwent reoperations. Clinical presentations, recurrence intervals, surgical findings, and clinical outcomes were analyzed. RESULTS: Analysis of 32 consecutive cases of recurrent TN initially attributable to veins revealed a female predominance (female/male = 26:5), with one female patient exhibiting bilateral TN caused by venous compression. Patient ages ranged from 15 to 80 years, with a prevalence in the seventh decade. The V2 distribution of the face was involved more frequently than other divisions. For 24 patients (75%), recurrence occurred within 1 year after the initial operation. At the time of the second MVD procedure, development of new veins around the nerve root was observed in 28 cases (87.5%). After successful subsequent MVD procedures, the pain was improved in 81.3% of the cases. CONCLUSION: The recurrence rate for TN attributable to veins is high. If pain recurs, it is likely to recur within 1 year after the initial operation. The most common cause of recurrence is the development and regrowth of new veins. Even fine new veins may cause pain recurrence; these veins may be located beneath the felt near the root entry zone or distally, near Meckel's cave. Because of the variable locations of vein recurrence, every effort must be made to identify recollateralized veins. Given the high rate of pain relief after a second operation, MVD remains the optimal treatment for the recurrence of TN attributable to vein regrowth. PMID- 10690725 TI - Percutaneous retrogasserian glycerol rhizolysis for treatment of chronic intractable cluster headaches: long-term results. AB - OBJECTIVE: To analyze the long-term effectiveness and safety of percutaneous retrogasserian glycerol rhizolysis (PRGR) in the treatment of medically refractive chronic cluster headache (CH). The current mainstay of surgical intervention for these patients is percutaneous radiofrequency retrogasserian rhizotomy (PRFR). However, when performed for V1 distribution pathology, PRFR can lead to corneal anesthesia, which places the patient at risk for future visual loss. It also increases the risk of facial dysesthesia. METHODS: In a prospective, consecutive series, 18 patients with intractable CH were followed for a mean of 5.2 years (range, 40-78 mo) after they had undergone PRGR, performed using a standard technique. The significance of this technique as an alternative to PRFR is that it should result in a lower rate of both corneal and facial anesthesia and provide an acceptable degree of pain relief. RESULTS: Fifteen patients (83%) obtained immediate pain relief after one or two injections; the majority of them experienced relief after the first injection. CH recurred in seven patients (39%) over the course of the study. Two of these patients received a second injection, and both met with equal success. Two other patients underwent PRFR. Excluding those who underwent PRFR, the overall daily headache frequency decreased from 3.5 +/- 0.3 attacks per day preoperatively to 0.6 +/- 0.2 attacks per day at last follow-up. The severity of these headaches, as assessed by verbal pain scales, also decreased from 10 preoperatively to 4.4 +/- 1.4 at follow-up. None of the patients, including those who required a second procedure, experienced corneal anesthesia or facial dysesthesia. CONCLUSION: This study provides the first long-term evaluation of PRGR for the treatment of medically refractive chronic CH and lends support to both the safety and long term efficacy of this procedure. Further investigations are needed to compare directly the relative efficacy and safety of PRGR and PRFR. PMID- 10690726 TI - Cowden disease and Lhermitte-Duclos disease: characterization of a new phakomatosis. AB - OBJECTIVE: Lhermitte-Duclos disease, or dysplastic gangliocytoma of the cerebellum, is an unusual hamartomatous lesion that can cause progressive mass effects in the posterior fossa. Cowden disease, or multiple hamartoma-neoplasia syndrome, is a rare autosomal dominant disorder characterized by mucocutaneous hamartomas and high incidences of systemic malignancies. We recently treated a patient with manifestations of both Lhermitte-Duclos disease and Cowden disease, and we were intrigued by the occurrence of these two rare disorders in the same patient. The purpose of the present study was to examine the nature of the association between Lhermitte-Duclos disease and Cowden disease. METHODS: The records for all patients who had been diagnosed at our institution as having Lhermitte-Duclos disease were reviewed, to determine whether these patients also exhibited manifestations of Cowden disease. Data were obtained from multiple sources, including patient interviews, correspondence with treating physicians, and chart reviews. RESULTS: During the past 40 years, five patients were diagnosed at Case Western Reserve University as having Lhermitte-Duclos disease. All five patients exhibited manifestations of Cowden disease. Before this review, Cowden disease had not been diagnosed for three of the patients. In our most recent case, the diagnoses of both disorders were established preoperatively. That patient was observed to have a deletion in the critical portion of Exon 5 of the PTEN gene, the gene associated with Cowden disease. CONCLUSION: Inclusion of Lhermitte-Duclos disease in the Cowden disease spectrum suggests that Cowden disease is a true phakomatosis, with hamartomas arising from cutaneous and neural ectoderm. Recent advances in molecular genetics may help to refine the current descriptive classification of the phakomatoses. The association between Lhermitte Duclos disease and Cowden disease has been under-recognized and under-reported. Recognition of this association has direct clinical relevance, because diligent long-term follow-up monitoring of individuals with Lhermitte-Duclos disease and Cowden disease may lead to the early detection of malignancy. PMID- 10690727 TI - Extreme lateral supracerebellar infratentorial approach to the posterolateral mesencephalon: technique and clinical experience. AB - OBJECTIVE: Lesions situated posterolaterally along the mesencephalon present neurosurgeons with a special challenge. The midline and paramedian variations of the supracerebellar infratentorial approaches do not adequately expose this region. The subtemporal approach risks injury to the vein of Labbe. An extreme lateral supracerebellar infratentorial approach with more radical resection of bone superiorly and laterally, and skeletonization of the sigmoid and transverse sinuses, was used to approach lesions at this location in eight METHODS: Five cavernous malformations, two juvenile pilocytic astrocytomas, and one peripheral superior cerebellar artery aneurysm located in this region were approached in eight patients. In this extreme lateral approach, the sigmoid sinus is unroofed more superiorly and the bone flap includes not only a posterior fossa craniotomy but also a portion that extends just above the transverse sinus. The dural opening is based along the transverse and sigmoid sinuses. After the cerebrospinal fluid has been drained, the lateral aspect of the brainstem is approached via the cerebellar surface. A proximal tentorial incision offers additional rostral exposure where needed. RESULTS: Seven patients in this series underwent successful resection of their lesion. The remaining patient's aneurysm was clipped successfully with no major complications. CONCLUSION: The extreme lateral supracerebellar infratentorial approach differs from the midline and paramedian supracerebellar infratentorial variants in the area of exposure, patient positioning, and location of the craniotomy. The technique is effective for approaching the posterolateral mesencephalon. PMID- 10690728 TI - Microelectrode-guided thalamotomy for Parkinson's disease. AB - OBJECTIVE: To describe the outcomes in our first 40 microelectrode-guided thalamotomies for parkinsonian tremor. METHODS: Twenty-four left-sided and 16 right-sided thalamotomies were performed between October 1984 and January 1996; the mean follow-up period was 35.8 months (range, 1-152 mo). The results were evaluated retrospectively and semiquantitatively by a disinterested observer (MNL) and correlated with the quality of the microelectrode recording and the number and size of radiofrequency lesions made. The first 20 and second 20 procedures were evaluated separately. RESULTS: At the last follow-up, the Unified Parkinson's Disease Rating Scale showed no or virtually no tremor in the upper limb in 75% of patients or in the lower limb in 73% of patients. No significant persistent complications were found. These results were achieved at the expense of having to repeat the procedure on 11 sides (in 5 because of technical problems and in 6 for no obvious reason). Total or nearly total abolition of tremor occurred after the first procedure in 40% of the first 20 operations and in 65% of the second 20. Eight of the first 20 procedures and 2 of the second 20 failed for technical reasons. Lesions were made larger in the second 20 procedures than in the first 20. With the use of an electrode with a 1.1 x 3-mm bare tip for 60 seconds, it seems that lesions had to be created at 60 degrees C or more to produce a successful result. CONCLUSION: Thalamotomy with microelectrode recording is an effective procedure with which to treat tremor in patients with Parkinson's disease and may involve fewer complications than conventional techniques. The procedure appears to involve a learning curve. PMID- 10690729 TI - Laminectomy versus percutaneous electrode placement for spinal cord stimulation. AB - OBJECTIVE: The purpose of this study was to compare the long-term effectiveness of spinal cord stimulation using laminectomy-style electrodes versus that using percutaneously implanted electrodes. METHODS: Forty-one patients underwent an initial trial period of spinal cord stimulation with temporary electrodes at Duke Medical Center between December 1992 and January 1998. A permanent system was implanted if trial stimulation reduced the patient's pain by more than 50%. Median long-term follow-up after permanent electrode placement was 34 months (range, 6-66 mo). Severity of pain was determined postoperatively by a disinterested third party using a visual analog scale and a modified outcome scale. RESULTS: Twenty-seven (66%) of the 41 patients participating in the trial had permanent electrodes placed. Visual analog scores decreased an average of 4.6 among patients in whom electrodes were placed via laminectomy in the thoracic region (two-tailed t test, P < 0.0001). Patients who underwent percutaneous placement of thoracic electrodes had an average decrease of 3.1 in their visual analog scores (two-tailed t test, P < 0.001). Electrodes placed through laminectomy furnished significantly greater long-term pain relief than did those placed percutaneously, as measured by a four-tier outcome grading scale (P = 0.02). CONCLUSION: Spinal cord stimulation is an effective treatment for chronic pain in the lower back and lower extremities that is refractory to conservative therapy. Electrodes placed via laminectomy in the thoracic region appear to be associated with significantly better long-term effectiveness than are electrodes placed percutaneously. PMID- 10690730 TI - Reevaluation of syringosubarachnoid shunt for syringomyelia with Chiari malformation. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness of syringosubarachnoid (S-S) shunting for syringomyelia with Chiari malformation. The authors describe the technical methods of performing the S-S shunt and the clinical results, including shunt malfunction. METHODS: Forty-nine patients underwent S-S shunting. These patients were divided into three groups according to differences in the surgical technique used. Group I patients underwent laminectomy plus midline myelotomy and had a shunt tube placed in the dorsal subarachnoid space. Group II patients underwent laminectomy plus dorsal root entry zone myelotomy and had a shunt tube placed in the dorsolateral subarachnoid space. Group III patients underwent hemilaminectomy plus dorsal root entry zone myelotomy and had a shunt tube placed in the ventrolateral subarachnoid space. RESULTS: Clinical results were generally satisfactory, especially in terms of pain relief, in all three groups. However, 10 patients among Groups I and II required follow-up surgery because of shunt problems; no second surgery was necessary for any patient in Group III. CONCLUSION: The S-S shunt was very effective in deflating the syrinx, and the clinical results were satisfactory. Therefore, even though foramen magnum decompression is a very effective treatment, S-S shunting should be reevaluated and not rejected; it should be considered as one of the major surgical options. To prevent the possibility of cord injury by myelotomy or shunt malfunction, the dorsal root entry zone should be selected as the myelotomy site, and the shunt tube should be inserted into the ventral subarachnoid space at the cervical level. PMID- 10690731 TI - A new technique for cranioplasty with L-shaped titanium plates and combination ceramic implants composed of hydroxyapatite and tricalcium phosphate (Ceratite). AB - OBJECTIVE: The use of hydroxyapatite-based ceramics for cranioplasties has recently increased in Japan, because of the good cosmetic outcomes, biocompatibility, strength, osteoconductive properties, and lack of risk of disease transmission associated with these materials. However, miniplate fixation has not been possible for ceramic implants. We describe a new technique for miniplate fixation of ceramic implants. METHODS: Combination ceramic implants composed of hydroxyapatite and tricalcium phosphate (Ceratite; NGK Spark Plug Co., Aichi, Japan) were used for cranioplasties. A slot and a pair of holes were cut in each Ceratite implant, for use as a fixation unit. We have also developed a new L-shaped titanium plate (HOMS Engineering Inc., Nagano, Japan) that fits into the fixation unit. We first insert an L-shaped titanium plate through the slot from the back surface of the Ceratite implant. We then bend the plate outward at the front surface of the Ceratite implant and fix it to the cranium of the patient with titanium screws. The Ceratite implant is usually firmly fixed to the cranium of the patient with three L-shaped titanium plates. RESULTS: Using L shaped titanium plates and Ceratite implants, we successfully performed cranioplasties for seven patients with cranial defects resulting from external decompression craniotomies. The Ceratite implant exactly fit the bone window for each patient. Surgical maneuvers were simple and easy for all patients, permitting shorter operating times. All Ceratite implants were firmly fixed, and no postoperative infections have occurred. CONCLUSION: Our new technique for cranioplasty is simple and allows rigid fixation of Ceratite implants. PMID- 10690732 TI - Retinoids inhibit human glioma cell proliferation and migration in primary cell cultures but not in established cell lines. AB - OBJECTIVE: Retinoids are known to exhibit a broad spectrum of biological activities, and they participate in the onset of differentiation and the inhibition of growth in a wide variety of cancer cells. Some of these vitamin A derivatives are already in clinical use. However, data on retinoid actions in glial tumors are rather sparse. Therefore, we studied the effects of the natural retinoic acid (RA) forms all-trans-RA, 9-cis-RA, and 13-cis-RA on glioma cell lines and primary cultures from patients with glioblastomas multiforme. METHODS: Six human glioma cell lines, one rat glioma cell line, and 20 primary cultures established from biopsies from patients with glioblastomas multiforme were investigated. Tumor cell proliferation was assessed using 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide and cell-counting assays. Random migration out of tumor spheroids was quantified using a video-morphometry system. Invasion was investigated using a confrontational coculture test system. Retinoid receptor (RA receptor [RAR]alpha, -beta, and -gamma and retinoid X receptor [RXR]alpha, beta, and -gamma) expression status was determined using reverse transcription polymerase chain reaction studies. RESULTS: Treatment of five human glioma cell lines with the different retinoids at concentrations up to 10(-5) mol/L produced no reduction of proliferation, using various incubation times. For one human glioma cell line (U343MG-A) and one rat glioma cell line (C6), which were previously reported to be sensitive to retinoids, we could confirm strong inhibitory effects on proliferation and clear changes in morphological features after retinoid treatment. Application of the different retinoids to low-passage primary cultures of human glioblastomas resulted in marked inhibition of proliferation (30-95%) for all tested samples. Using three-dimensional spheroid cultures, we detected retinoid-induced decreases in cell migration (24-65%). Invasion was not affected by these vitamin A derivatives. In an analysis of the expression patterns for retinoid receptors (RARs and RXRs), all primary culture samples yielded positive results for RAR gamma and RXR alpha and negative results for RAR alpha, RAR beta, and RXR gamma, whereas the results of RXR beta expression were heterogeneous among different patients. The cell lines, irrespective of their RA sensitivities, did not exhibit any major differences in receptor expression. CONCLUSION: Retinoids strongly inhibit proliferation and migration in primary cultures of human glioblastomas multiforme. Our data support a clinical trial of retinoids for the treatment of human malignant gliomas. We observed that most established cell lines were not sensitive to RA. This difference between long-term cell lines and primary cultures cannot be explained by different retinoid receptor expression patterns. PMID- 10690733 TI - Induction of apoptosis in glioma cells by recombinant human Fas ligand. AB - OBJECTIVE: Fas ligand (FasL) belongs to the tumor necrosis factor family and has the ability to induce apoptosis in susceptible target cells by binding to its receptor, Fas. It has been demonstrated recently that the FasL/Fas system plays a pivotal role in the cytocidal activity of T lymphocytes in the immune system. FasL may act as a cytotoxic effector molecule to Fas-expressing malignant tumor cells. We reported previously that Fas is commonly expressed in human brain tumor cells. In this study, we examine the possible application of FasL to therapy for malignant brain tumors. METHODS: To develop an expression system yielding large amounts of FasL, we constructed a baculovirus vector containing complementary deoxyribonucleic acid of human FasL under the control of a polyhedrin promoter. We produced human FasL in Spodoptera frugiperda (Sf9) insect cells infected by the recombinant baculovirus carrying FasL complementary deoxyribonucleic acid and studied the cytocidal activity of FasL against the T98G human glioblastoma cell line. RESULTS: FasL expression in Sf9 cells was confirmed immunocytochemically with rabbit antibody raised against the cytoplasmic domain of human FasL. The FasL released into the supernatant of cultured Sf9 cells was also verified by Western blotting, and it specifically induced apoptosis in T98G cells. The induced apoptosis by recombinant human FasL was confirmed by annexin V fluorescein isothiocyanate staining. CONCLUSION: The present results suggest that the induction of apoptosis by the Fas/FasL system could be a new strategy for the treatment of malignant brain tumors, which are resistant to conventional therapies. PMID- 10690734 TI - Thin and thick filament regulation of contractility in experimental cerebral vasospasm. AB - OBJECTIVE: Cerebral vasospasm is a potentially fatal consequence of aneurysmal subarachnoid hemorrhage and influences the prognosis of the patient. The purpose of this study was to evaluate the status of thin (actin) and thick (myosin) filament regulation of smooth muscle contraction in the double-subarachnoid hemorrhage canine model of cerebral vasospasm and to determine the effects of a kinase inhibitor reported to be effective in vasospasm, HA1077, on thin and thick filament regulation. METHODS: Cerebral vasospasm was assessed by vertebral angiography. Myosin regulatory light chain phosphorylation was measured using glycerol-urea gels, whereas protein levels of the thin filament-associated protein calponin were measured by Western blot. RESULTS: The basilar arteries of dogs in which subarachnoid hemorrhage was induced narrowed to 36% +/- 2.0% of their size on the first day (n = 12). The phosphorylation of the regulatory light chain tended to increase, but the change did not reach statistical significance (35% +/- 5.9% [n = 12] versus 25% +/- 4.8% [n = 10] in control arteries). In contrast to this increase, significant degradation of calponin was observed in the samples from vasospastic dogs (85.4% +/- 5.45% [n = 5] versus 15.2% +/- 6.21% [n = 5]; P < 0.01). Prophylactic treatment with intravenous injections of HA1077 at 0.67 mg/kg b.i.d. significantly inhibited vasospasm (diameters, 65% +/- 10.2% of Day 1 diameters [n = 5]; P < 0.05), and calponin degradation (57.8% +/- 13.9% [n = 4]) was substantially reduced. CONCLUSION: These data suggest that degradation of the thin filament-associated protein calponin plays a role in cerebral vasospasm and that the antivasospastic action of HA1077 is, at least in part, due to prevention of calponin degradation. PMID- 10690735 TI - In vivo animal models of cerebral vasospasm: a review. AB - BACKGROUND: Cerebral vasospasm is delayed-onset cerebral arterial narrowing in response to blood clots left in the subarachnoid space after spontaneous aneurysmal subarachnoid hemorrhage (SAH). Ideally, studies on the pathogenesis and treatment of cerebral vasospasm in humans should be conducted using human cerebral arteries. Because in vivo experiments using human vessels are not possible, and postmortem pathological examination of human arteries in vasospasm provides only a limited amount of information, a number of animal models of vasospasm have been developed. METHODS: The literature was searched to find all references to in vivo animal models of SAH and vasospasm. An online search of the medical database MEDLINE was initially performed using the key words "cerebral," "vasospasm," "subarachnoid," "hemorrhage," "animal," and "model." References were checked to determine the first description of each in vivo animal model. RESULTS: Fifty-seven models of SAH and vasospasm were identified. These models used one of three techniques to simulate SAH: 1) an artery was punctured allowing blood to escape and collect around the artery and its neighbors; 2) an artery was surgically exposed, and autologous blood obtained from another site was placed around the artery; or 3) blood from another site was injected into the subarachnoid space and was allowed to collect around arteries. Each technique has advantages and disadvantages. The majority of animal models of SAH and vasospasm use intracranial arteries; however, extracranial arteries have also been used recently in vasospasm experiments. These studies seem easier and less costly to perform, but concerns exist regarding the physiological dissimilarity between systemic and cerebral arteries. CONCLUSION: The model of SAH and vasospasm used most frequently is the canine "two-hemorrhage" model, in which two injections of blood into the dog's basal cistern performed 48 hours apart result in greater arterial vasoconstriction than that effected by a single injection of blood. On the basis of its ability to accurately predict what occurs in human SAH, the best model of vasospasm seems to be the primate model in which a blood clot is surgically placed around the large cerebral vessels at the base of the monkey's brain. PMID- 10690736 TI - A tribute to Dr. Fedor A. Serbinenko, founder of endovascular neurosurgery. AB - From humble beginnings in the former Soviet Union, Fedor A. Serbinenko, M.D., Ph.D., became a leading figure at Moscow's famed Burdenko Neurosurgery Institute. While there, he invented and perfected the technique of balloon embolization, which was destined to change the practice of neurovascular surgery forever. We present the life and achievements of the father of endovascular neurosurgery. PMID- 10690737 TI - Origin and evolution of the Department of Neurosurgery, Neurological Institute, Faculty of Medicine, Kyushu University. AB - The Department of Neurosurgery at Kyushu University had its origins within the First Department of Surgery and was established as a subspecialty at the Neurological Institute more than 30 years ago under the leadership of Katsutoshi Kitamura. Further development of the neurosurgical department has proceeded during the chairmanship of Masashi Fukui. These leaders and many other dedicated physicians and surgeons, nurses, investigators from other countries, and staff members have contributed to the creation of a research-oriented neurosurgical environment that interacts fruitfully with the other components of the Neurological Institute. This article describes the development of neurosurgery within Kyushu, which has been a highly cosmopolitan area throughout its long history. More specifically, this account outlines the origin and growth of the Department of Neurosurgery at Kyushu University. PMID- 10690738 TI - Isolated cerebellar involvement in Rosai-Dorfman disease: case report. AB - OBJECTIVE AND IMPORTANCE: Sinus histiocytosis or Rosai-Dorfman disease (RDD) is a rare but well-recognized disorder characterized by an unusual proliferation of histiocytic cells. Intracranial localization is a rare manifestation of RDD. Only three cases of localization in the posterior fossa have been reported in the literature. The present report describes the first case, to our knowledge, of cerebellar localization of RDD. CLINICAL PRESENTATION: A 67-year-old woman was admitted to our institution with a 5-month history of cerebellar ataxia. Her medical history was unremarkable. The patient was alert and cooperative. No cranial nerve deficits were evident; Romberg positivity to the left side was recorded. No cutaneous abnormalities, lymphadenopathy, or hepatosplenomegaly were revealed by physical examination. Routine hematological and biochemical studies were normal except for the erythrocyte sedimentation rate, which was elevated. Radiologically, the lesion appeared as a well-defined and avascular mass in the right cerebellar lobe. Meningioma was considered the most likely diagnosis. TECHNIQUE: The patient underwent a suboccipital craniotomy with complete excision of the lesion. Microscopic examination of the operative specimen revealed the presence of a mixed cellular population with predominant mature histiocytes. A peculiar feature was the presence of lymphocytes and monocytes within the cytoplasm of histiocytes (emperipolesis). Immunohistochemical study of the histiocytes revealed strong positivity for S-100, CD-68 antigen, and vimentin. CONCLUSION: Involvement of the central nervous system in RDD appears to have a benign prognosis, especially in the absence of nodal diseases. Surgery is essential for diagnosis, and, when total removal is achieved, the outcome is generally good without risk of recurrence. PMID- 10690739 TI - Radiographic evidence and surgical confirmation of a saccular aneurysm on a hypoplastic duplicated A1 segment of the anterior cerebral artery: case report. AB - OBJECTIVE AND IMPORTANCE: True duplication of the A1 segment of the anterior cerebral artery is extremely rare, as is finding a true A1 segment saccular aneurysm. We report the angiographic and surgical findings of such a case with the additional association of a hypoplastic ipsilateral M1 segment of the middle cerebral artery. CLINICAL PRESENTATION: A 68-year-old man presented with a Hunt and Hess Grade II subarachnoid hemorrhage and symptoms of headache, nuchal rigidity, and facial paresis. INTERVENTION: Angiographic evaluation with superselective exploration revealed a small ruptured aneurysm located on a duplicated hypoplastic A1 segment of the left anterior cerebral artery with associated middle cerebral artery stenosis and secondary early moyamoya changes. Surgical clipping of the aneurysm was performed successfully while sparing the hypoplastic A1 segment. CONCLUSION: A1 aneurysms occurring on a duplicated anterior cerebral artery segment probably develop from a congenital weakness of the parent vessel and increased local shear stress. Superselective angiography was helpful in the preoperative planning and facilitated the decision to treat with surgical clipping instead of embolization. PMID- 10690740 TI - Spontaneous spinal cord herniation: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Spontaneous herniation of the spinal cord substance through a previously uninjured and/or untouched dura is a very exceptional occurrence. Spontaneous spinal cord herniation, which was first reported 25 years ago, is a cause of myelopathy that is treatable but difficult to diagnose. CLINICAL PRESENTATION: A 49-year-old female patient who presented with a 3-year history of a burning sensation and hyperesthesia in her right leg and a 3-month history of left leg stiffness was diagnosed as exhibiting signs of Brown-Sequard syndrome. Magnetic resonance imaging of the thoracic spinal canal demonstrated S shaped anterior kinking and transdural herniation of the spinal cord at the T3-T4 levels. INTERVENTION: The patient underwent surgery via a three-level laminectomy. The herniated part of the spinal cord was microsurgically reduced, and the dural defect was repaired with Gore-Tex membrane (WL Gore & Associates, Flagstaff, AZ). The outcome of surgery was excellent. CONCLUSION: Review of the world literature revealed 29 reported cases, with 27 of these cases being published since 1990. The clinical features, radiological diagnosis, and treatment options for this unique entity are summarized, with a synopsis of numerous misconceptions that appeared in the literature. With more familiarity with and increased awareness of this entity, more cases will be diagnosed. PMID- 10690741 TI - Progressive spontaneous herniation of the thoracic spinal cord: case report. AB - OBJECTIVE AND IMPORTANCE: We report one case of spontaneous thoracic spinal cord herniation. To our knowledge, this is the first case involving radiological documentation of the development of herniation. Clinical features and surgical techniques are also presented. CLINICAL PRESENTATION: We describe the case of a 51-year-old female patient who experienced progressive Brown-Sequard syndrome for 2 years. Three magnetic resonance imaging examinations were performed; they revealed the progressive development of anterolateral spinal cord herniation at the level of T6 during those 2 years. INTERVENTION: After laminectomy at T6, the herniated myelon was microsurgically removed and the neurological symptoms improved. CONCLUSION: We present the possible causes, the proposed pathophysiological mechanisms, and the clinical and radiological development of this rare entity, with a review of the literature published to date. We propose that a preexisting weakness of the ventral dural fibers, combined with abnormal adhesion of the spinal cord to the anterior dural sleeve, leads to progressive herniation throughout life. Microsurgical treatment may halt the exacerbation of the neurological symptoms. PMID- 10690742 TI - Endovascular treatment of a grade IV transverse sinus dural arteriovenous fistula by sinus recanalization, angioplasty, and stent placement: technical case report. AB - OBJECTIVE AND IMPORTANCE: The frequent association of dural arteriovenous fistulae (DAVFs) and dural sinus thrombosis may render the treatment of these complex lesions difficult. We report a case of DAVF eradicated by recanalization of the chronically thrombosed transverse sinus (TS) and sigmoid sinus followed by balloon angioplasty and stent deployment at the site of the fistula. CLINICAL PRESENTATION: A 52-year-old man presented with a Type IV DAVF of the left TS with widespread white matter changes secondary to venous hypertension. Arterial feeders arose from the left internal carotid, external carotid, and vertebral arteries. The distal segment of the left TS, the left sigmoid sinus, and the proximal segment of the right TS were occluded. Reverse flow was observed in the deep venous system and in the superior sagittal sinus. INTERVENTION: Endovascular access was gained through the left internal jugular vein. Mechanical recanalization of the thrombosed left TS and sigmoid sinus was followed by balloon angioplasty and placement of six overlapping stents extending from the TS to the proximal internal jugular vein. Angiograms performed after surgery showed resaturation of antegrade venous drainage as well as complete eradication of the fistulous connections. The patient was discharged with an improving clinical CONCLUSION: Recanalization of a chronically occluded dural venous sinus through a jugular approach is feasible. In addition to eradicating cerebral venous hypertension by reestablishing antegrade venous drainage, balloon angioplasty and stent deployment at the DAVF site produced complete closure of the fistula. This may prove to be a new therapeutic strategy for management of DAVF. PMID- 10690743 TI - Navigus trajectory guide. PMID- 10690744 TI - Prognostic value and determinants of ultraearly angiographic vasospasm after aneurysmal subarachnoid hemorrhage. PMID- 10690745 TI - Patients with spinal cord cavernous malformations are at an increased risk for multiple neuraxis cavernous malformations. PMID- 10690746 TI - Stereotactic targeting of the globus pallidus internus for Parkinson's disease: imaging versus electrophysiological mapping. PMID- 10690747 TI - History of the operating microscope: from magnifying glass to microneurosurgery. PMID- 10690748 TI - Flurothyl-induced seizures in rats activate Fos in brainstem catecholaminergic neurons. AB - Autonomic changes accompany seizures in both animals and humans. While ictal autonomic dysfunction can be life-threatening, the participating neural networks involved are poorly understood. In this study we examined the activation of Fos following generalized seizures in brainstem structures known to mediate autonomic function. Adult female rats were sacrificed 2 h after flurothyl-induced seizures. Double-immunostaining for c-Fos and dopamine-beta-hydroxylase (DBH), and c-Fos and phenylethanol-N-methyl-transferase (PNMT) were performed in brainstem slices. Numbers of DBH-labeled neurons expressing Fos-like immunoreactivity (FLI) (DBH/Fos) and PNMT labeled neurons expressing FLI (PNMT/Fos) were counted in the noradrenergic (A1, A2, A5, A7) and adrenergic (C1, C2) cell groups localized in pons and medulla oblongata. Among the experimental animals, the highest degree of co-localization of DBH/Fos neurons was observed in the locus coeruleus (A6; 87.7%), and in the A1(72.8%) cell group located in the caudal ventrolateral medulla (VLM). No co-localization of DBH/Fos neurons was observed in control animals. The highest degree of co-localization of PNMT/Fos neurons was observed in the C1 adrenergic cell group (84.2%) located in the rostral VLM. Control animals showed very few (5.5%) PNMT/Fos co-localized neurons in the C1 adrenergic cell group. Our results indicate that flurothyl-induced generalized seizures in rats activate catecholaminergic neurons in the pons and medulla oblongata. Further studies are necessary to determine whether activation of brainstem catecholaminergic neurons contribute to the autonomic manifestations that frequently accompany epileptic seizures. PMID- 10690749 TI - Neocortical hyperexcitability after GABA withdrawal in vitro. AB - The sharp interruption of the intracortical instillation of exogenous gamma aminobutyric acid (GABA), generates an epileptic focus in mammals. Seizures elicited by GABA withdrawal last several days or weeks. The present work reports that GABA withdrawal-induced hyperexcitability can be produced in vitro: a sudden withdrawal of GABA (5 mM; 120 min) or benzodiazepine (60 microM flunitrazepam) from the superfusion, induced a gradual increase in the amplitude of the evoked population spike (PS) recorded on neocortical slices. PS enhancement reached 150% above the control value 2.5 h after GABA withdrawal. GABA withdrawal-induced hyperexcitability was facilitated by progesterone. PS enhancement induced by GABA withdrawal was associated with an impairment of GABA transmission occurring before epileptiform discharges were fully established. Paired pulse inhibition and evoked [3H]-GABA release appear decreased; suggesting that cortical hyperexcitability as a result of GABA withdrawal involves pre-synaptic changes. Specific muscimol binding decreased during GABA superfusion but recovered after GABA withdrawal. However, the sensitivity of the post-synaptic response to 3alpha OH-5alpha-pregnan-20-one or allopregnanolone (alloP) was enhanced after GABA withdrawal, suggesting a functional change in the GABA(A) receptors. The changes described may be the cellular correlates of the withdrawal syndromes appearing after interruption of the administration of GABA(A) receptor agonists. PMID- 10690750 TI - No association found between polymorphisms in genes encoding mGluR7 and mGluR8 and idiopathic generalised epilepsy in a case control study. AB - The genes of two group III metabotropic glutamate receptors, mGluR7 and 8, are candidate susceptibility genes for epilepsy. The Tyr433Phe polymorphism of mGluR7 and a novel polymorphism in the mGluR8 gene located 29 bp after the termination codon (2756C/T) were investigated in case control association studies performed on DNA from more than 100 patients with idiopathic generalised epilepsy (IGE). No significant association was found with IGE for either polymorphism. PMID- 10690751 TI - Running down phenomenon and acute interictal psychosis in medial temporal lobe epilepsy. AB - PURPOSE: To examine the relationship between presurgical acute interictal psychosis and postsurgical running down phenomenon in a strictly homogeneous group with medial temporal lobe epilepsy. METHODS: Forty patients with mesial temporal sclerosis and an ultimate excellent surgical outcome, were divided into running down positive and negative groups. Various clinical and laboratory data, including presurgical psychotic episodes, were analyzed in the two groups. RESULTS: Patients in the running down positive group exhibited a significantly high incidence of presurgical history of acute interictal psychosis. CONCLUSION: We hypothesize that areas of secondary epileptogenesis beyond the scope of the primary epileptogenic zone, suggested by postsurgical running down phenomenon, may play an important role in the blockage and shift of the habitual propagation of seizure activity from the primary epileptogenic zone, to which Wolf attributed acute interictal psychosis. PMID- 10690752 TI - Acute behavioral and EEG effects of NW-1015 on electrically-induced afterdischarge in conscious monkeys. AB - NW-1015 is a novel Na+ and Ca2+ channel blocker with broad spectrum anticonvulsant activity and an excellent safety margin. As the compound also shows sigma-1 receptor ligand properties it was deemed important to determine whether it possesses anticonvulsant properties in primates without causing behavioral and EEG abnormalities. Thus, the effects of NW-1015 on limbic electrically-induced afterdischarge (AD) were evaluated in four cynomolgus monkeys, and its activity compared to a single effective dose of phenytoin (PHT). The four male cynomolgus monkeys were chronically implanted for EEG recordings, from cortex and limbic structures. AD was induced in limbic areas by electrical stimulation. The effects of NW-1015 on the duration and the behavioral component of the AD were randomly tested at doses from 25 to 75 mg/kg and compared with the effects of PHT 50 mg/kg. Similarly to PHT, 50 mg/kg of NW-1015 significantly shortened the EEG AD and almost abolished AD elicited behavioral seizure. Only the behavioral effects of AD were reduced after administration of 25 mg/kg p.o. NW-1015 did not cause EEG or interictal behavioral alterations at doses up to 75 mg/kg p.o. These data further confirm the broad-spectrum anticonvulsant activity and a good safety profile of NW-1015 even in a primate model of complex partial seizures and suggest that its affinity for sigma-1 receptors is behaviorally irrelevant. PMID- 10690753 TI - Felbamate block of recombinant N-methyl-D-aspartate receptors: selectivity for the NR2B subunit. AB - The anticonvulsant felbamate blocks N-methyl-D-asparate (NMDA) receptors but fails to exhibit the neurobehavioral toxicity characteristic of other NMDA receptor antagonists. To investigate the possibility that felbamate's favorable toxicity profile could be related to NMDA receptor subtype selectivity, we examined the specificity of felbamate block of recombinant NMDA receptors composed of the NR1a subunit and various NR2 subunits. Felbamate produced a rapid, concentration-dependent block of currents evoked by 50 microM NMDA and 10 microM glycine in human embryonic kidney 293 cells expressing the rat NR1a subunit, and either the NR2A, NR2B or NR2C subunits; the IC50 values for block were 2.6, 0.52 and 2.4 mM, respectively (holding potential, - 60 mV). The Hill coefficient values were < 1 and, in kinetic analyses, onset and recovery from block were well fit by double exponential functions, indicating binding to more than one blocking site on the NMDA receptor channel complex. The higher affinity of felbamate block of NMDA receptors containing the NR2B subunit could be accounted for by more rapid association and slower dissociation from these sites. We conclude that felbamate exhibits modest selectivity for NMDA receptors composed of NR1a/NR2B subunits. This selectivity could, in part, account for the more favorable clinical profile of felbamate in comparison with NMDA receptor antagonists that do not show subunit selectivity. PMID- 10690754 TI - Genetic variation of the human mu-opioid receptor and susceptibility to idiopathic absence epilepsy. AB - Pharmacological and autoradiological studies suggest that mu-opioid receptor (OPRM) mediated neurotransmission is involved in the generation of absence seizures. Mutation screening of the human OPRM gene identified a common amino acid substitution polymorphism (Asn40Asp) that differentially modulates the binding affinity of beta-endorphin and signal transduction of the receptor. The present association study tested the candidate gene hypothesis that the Asn40Asp substitution polymorphism in the N-terminal OPRM domain confers genetic susceptibility to idiopathic absence epilepsy (IAE). The genotypes of the Asn40Asp polymorphism were assessed by allele-specific polymerase chain reaction in 72 German IAE patients and in 340 ethnically matched control subjects. The frequency of the Asp40 allele was significantly increased in the IAE patients [f(Asp40) = 0.139] compared to the controls [f(Asp40) = 0.078; chi2 = 5.467, df = 1, P = 0.019; OR = 2.03; 95%-CI: 1.12-3.68]. This allelic association suggests that the functional Asp40 variant of OPRM modulates neuronal excitability underlying the epileptogenesis of IAE. PMID- 10690755 TI - CNS oxidative stress associated with the kainic acid rodent model of experimental epilepsy. AB - The role of oxidative stress in seizure-induced brain injury was investigated in a kainic acid model of experimental epilepsy. Kainic acid (12.5 mg/kg) or saline was injected intraperitoneally into 12-week-old male Fischer 344 rats and sacrificed by decapitation at 4 and 24 h after injection. Markers of oxidative stress including protein carbonyls, thiobarbituric acid reactive material (TBARs), glutathione (GSH) and glutathione disulfide (GSSG) were measured in hippocampus, cortex, cerebellum and basal ganglia. Four hours after treatment, protein carbonyls were elevated by 103, 55, 52 and 32% in cortex, hippocampus, basal ganglia and cerebellum, respectively. TBARs were increased by 30-45% in all areas. After 24 h, elevated protein and lipid oxidative markers persisted in the hippocampus and cerebellum; by contrast, in the cortex, TBARs almost normalized to control values and protein carbonyls trended downward by one-half compared with measurements at 4 h, although this reduction relative to the 4 h timepoint did not reach statistical significance. In the basal ganglia, protein carbonyls approached control values at 24 h. GSSG levels were only increased statistically in the cortex after 4 h, GSH levels in all the regions were unchanged after treatment with kainic acid. However, in cortex, GSH levels correlated negatively with increases in protein and lipid oxidation (r = -0.69, P < 0.002). In contrast, significant correlations between GSH, protein carbonyls and TBARs measured in the hippocampus or cerebellum were not observed. Our data suggests that kainic acid induced similar oxidative stress in all of the brain regions that were examined, and that GSH plays a major antioxidant role in the cerebral cortex but not the hippocampus. PMID- 10690756 TI - A questionnaire survey about doctor-patient communication, compliance and locus of control among south Indian people with epilepsy. AB - To date, very few studies have investigated patients' views on the information they receive from the doctor concerning epilepsy and its management. Little information is available about the influence of doctor-patient communication and locus of control on the compliance of persons with epilepsy. We investigated, through a questionnaire-interview design, among patients attending the epilepsy clinic of a tertiary referral center in South India, their views about the provision of information by the doctor, and their compliance and locus of control. We also determined the interrelation between doctor-patient communication, compliance and locus of control. Our subjects comprised 200 adult persons with epilepsy, 113 males and 87 females, mean age 30.5 (range 18-67) years. Over one-third of the subjects received from the doctor insufficient information about epilepsy and its treatment. There was a significant positive correlation between effective doctor-patient communication and compliance. A majority of our patients had an external locus of control, which negatively influenced the compliance. Even in a comprehensive epilepsy clinic of a model tertiary referral center in a developing country, a significant proportion of patients do not receive optimal information about epilepsy from the doctor. Knowledge about their disease will encourage people with epilepsy to make informed choices, and achieve better compliance and personal control of their problems. Educating primary and secondary care physicians about the importance of doctor-patient communication in the management of epilepsy and educating the public about the positive aspects of life in epilepsy cannot be overemphasized. PMID- 10690757 TI - Report of the Coding Committee of the Association for European Paediatric Cardiology. PMID- 10690758 TI - The European Paediatric Cardiac Code Long List: structure and function. PMID- 10690759 TI - Inhibitors on an elastase-like enzyme activity catalyzing Suc-Ala-Ala-Pro-Leu-pNA amidolysis in human seminal plasma. AB - The behavior of some proteinase inhibitors toward the Suc-Ala-Ala-Pro-Leu-pNA amidolytic enzyme activity in human seminal plasma (HSP) was tested. [(2S, 3R)-3 Amino-2-hydroxy-5-methyl-hexanoyl]L-valyl-L-valyl-L-aspartic acid (Amastatin) and 3-[1-[(2-(hydroxymethyl)- -pyrolidinyl)-2-methylpropyl]-carbamoyl] octanohydroxamic acid (Actinonin) showed strong inhibitory effects. No inhibition of this present enzyme activity was seen with anti-human serum (whole), anti human leukocyte elastase, phenyl-methyl sulfonyl fluoride, Elastatinal, ethyeneglycol bis(beta-aminoethyl ethyl)N,N,N:N'-tetra acetic acid, and [L-3 trans-ethoxycarbonyl-oxirane-2-carbonyl]1-L-leucine(3-methylbutyl)a mido (E-64). No relation was observed between human pancreatic elastase antigen and the Suc Ala-Ala-Pro-Leu-pNA amidolytic enzyme enzyme activity in HSP. Two peaks of Suc Ala-Ala-Leu-Pro-pNA amidolytic enzyme activity were separated by Cellulofine GCL 2000 gel filtration and these activities were completely abolished by addition of Amastatin. Suc-Ala-Ala-Pro-Leu-pNA amidolytic enzyme activity in HSP is not an elastase-like metalloproteinase but is rather an acyl amidase-like leucine aminopeptidase. PMID- 10690760 TI - Effect of immunization with synthetic peptide corresponding to region 1-17 of human seminal plasma inhibin on fertility of male rats. AB - Immunization of adult male rats with a synthetic peptide corresponding to the region 1-17 of human seminal plasma inhibin (hSPI) resulted in agglutination of epididymal sperm, severely affecting the fertility of the animals (75% reduction in fertility as compared to control). This effect was found to be dependent on the antibody titer to hSPI. There was a significant rise in circulating follicle stimulating hormone levels, with luteinizing hormone and testosterone levels remaining unaffected. The histology of the testes and other reproductive organs revealed that these organs remained unaltered. The N-terminal 1-17 amino acid peptide of hSPI may hold promise as an immunogen for male immunocontraception. PMID- 10690761 TI - Antioxidative effect of melatonin on human spermatozoa. AB - The ability of melatonin to suppress experimentally induced lipid peroxidation (LPO) in sperm membrane was investigated in 41 samples of infertile men. Iron/ascorbate (0.04/0.2 mmol)-induced LPO was measured by the formation of malondialdehyde (MDA) using the thiobarbituric acid method. Sperm incubated in the presence of melatonin (2-6 mmol) exhibited a concentration-dependent decrease of MDA generated from hydroperoxide of the sperm plasma membrane in the presence of promoter system. Addition of 6 mmol of melatonin significantly reduced the rate of lipid peroxidation in sperm of unselected donors (mean +/- SE in control samples = 26.4 +/- 2.9 vs. 6.5 +/- 1.1 nmol MDA/10(8) sperm in melatonin-treated samples: n = 16, p < .005). Inhibitory effect of melatonin was also significant in the presence of 0.015 mmol of ferrous ions (20.5 +/- 1.7 vs. 7.9 +/- 1.6 nmol MDA/10(8) sperm in melatonin-treated samples: n = 7, p < .02) and (.005 mmol of ferrous ions (20.2 +/- 2.8 vs. 9.9 +/- 2.4 nmol MDA/ 10(8) sperm: n = 6, p < .05). Comparing the effect of melatonin with that of Trolox, an analog of vitamin E. a similar effect at concentration of 0.1-0.2 mmol of Trolox was found (25.2 +/ 2.9 vs. 11.8 +/- 1.2 nmol MDA/10(8) sperm in Trolox-treated samples: n = 7, p < .005). The obtained data of in vitro experiments show that melatonin is 40-fold less efficient than Trolox in achieving the 50% reduction in LPO (4 vs. 0.1 mmol). Since the physiological concentration of melatonin in human semen is at the nanomolar level, its antioxidative role in vivo is probably of minor importance. PMID- 10690762 TI - Autostage sperm tracing system for semen evaluation. AB - To overcome the limitation of the microscope field, the study proposed an autostage sperm tracing system (ASTS), which could trace a particular sperm for a long time and distance. The ASTS was constructed by assembling a commercial microscope, an image frame grabber, a personal computer, and a motorized stage. Its performance was tested by evaluating 6 semen samples and by comparing the evaluation with those of other semen evaluations. The ASTS broke through the limitation of the microscope field and traced a particular sperm as long as possible. It analyzed the sperm track and calculated the motility parameters, such as curvilinear velocity (Vcl), straight-line velocity (Vsl), and linearity (L(in)). The sperm quality was then evaluated in real time, and the user could decide to capture or abandon a particular sperm in the IVF The ASTS enables users to evaluate sperm progression for a long time and to have the global quality of a particular sperm in real time. Its open structure has the flexibility for micromanipulating a semen sample, and has the potential application associated with a modern IVF technique. PMID- 10690763 TI - A study of to determine if limiting the contact of sperm with zona pellucida reduces the rate of spontaneous abortions. AB - A recent study suggested that oligoasthenozoospermia may be an etiologic factor for spontaneous abortion (SAB) after in vitro fertilization-embryo transfer (IVF ET). However, IVF-ET with intracytoplasmic sperm injection (ICSI) did not seem to be associated with an increased SAB rate. The study presented herein compared the rate of SAB in pregnancies achieved by IVF-ET according to the type of oocyte insemination process. The 3 types evaluated were conventional insemination, which exposed the oocyte to 25,000 sperm with prolonged contact (16-24 h), intermediate contact with a short insemination protocol where contact with 25,000 sperm was limited to 2 h, and very limited contact with ICSI, where only 1 sperm was injected into the oocyte thus not exposing the zona pellucida to any sperm. The patients were further subdivided into age groups of < or =39 or > or =40. SAB rates after frozen ET were also evaluated. The clinical pregnancy and SAB rates following fresh or frozen ET for conventional, ICSI, and short insemination techniques for the 2 age groups were comparable. These data question whether oligoasthenozoospermia may be a factor in causing SAB, and whether avoidance of contact with the zona pellucida by using ICSI can negate this effect. A larger study is needed. PMID- 10690764 TI - Separation of sperm through a 12-layer percoll column decreases the percentage of sperm staining with quinacrine. AB - Previous methods of enriching sperm with a higher percentage of Y-bearing sperm have been questioned because the claims that Y enrichment was present were based on quinacrine staining of the Y chromosome, and the enrichment was not confirmed by polymerase chain reaction (PCR) or fluorescent in situ hybridization (FISH) techniques. A technique was evaluated that theoretically could increase the percentage of X-bearing sperm by isolating a fraction of the "heaviest" sperm by passing them through 12 layers of discontinuous Percoll gradient. Initially 12 specimens were checked both before and then after separation with 12 layers of Percoll for percentage of Y sperm. The median for baseline Y percentage was 49% and after processing the percentage of Y dropped to 10%. An additional 19 specimens were checked after separation only. The median was 19%. The sample with the lowest preseparation % of quinacrine staining sperm was 45% and the highest was 54%. After 12-layer Percoll, the lowest percentage was 3% and the highest was 24%. There have been claims that quinacrine staining can falsely increase apparent Y-bearing sperm enrichment following certain separation procedures, e.g.. albumin separation, by nonspecific staining of autosomal chromosomes. If anything, then, it should falsely decrease X-bearing sperm enrichment. Thus, 12 layer Percoll separation may actually enrich for X-bearing sperm or possibly this procedure somehow nonspecifically inhibits the ability of quinacrine to stain the Y chromosome. PMID- 10690765 TI - Ultrastructural investigation of human sperm using atomic force microscopy. AB - Ultrastructural investigation of human sperm in its natural environment (without fixation, dehydration, embedding, sectioning, etc.) was carried out by using atomic force microscope (AFM) in its tapping mode. This technique permits the examination of fine structural details of undamaged sperm and its topography with precision. Moreover, it allows 3D reconstruction of images and enhances the contrast to resolve details such as mitochondria that surround the axoneme at the sperm middle piece. An organized structure has been found in the flageller axoneme region. Ultrastructure also reveals folding and details of the depression of the membrane that cannot be examined with conventional techniques. PMID- 10690766 TI - A modified acrosome induction test. AB - Many types of acrosome induction tests require special equipment and reagents that are not available to most clinicians; thus, simpler tests seem desirable. A modified acrosome induction test has been developed that uses basic reagents and a light microscope, which are available in most office settings. A hypoosmotic swelling test and a double stain (Bismark brown and rose Bengal) were combined to evaluate the viable acrosome reaction (AR) among 74 infertile men and 42 control men. The study included 34 infertile males without varicoceles, 20 with nonrepaired varicoceles and 20 with repaired varicoceles. On each test day, a specimen from a fertile donor was run as a control. The spontaneous acrosome reaction was recorded in semen before and after capacitation. The final % viable acrosome reaction equaled the capacitated value minus the spontaneous value for whole semen. The mean % viable AR among the control specimens was 16% with no values less than 10%. This mean value for controls was significantly greater than the mean % viable AR in each patient group. There were no overlaps in the 95% confidence intervals. When the study group was stratified according to normal acrosome induction tests or >10% viable AR, 30 patients had a normal test and 44 had abnormal tests. Six patients with varicoceles and an abnormal acrosome induction test had a varicocelectomy, and 2 (33%) converted their acrosome induction test to normal after at least 6 months of follow-up. Nine patients had in vitro fertilization (IVF), 3 had a poor result, and all had an abnormal acrosome induction test. Six had a good result with IVF and all 6 had a normal acrosome induction test. Thus, the acrosome induction test described in this report may be performed in any office laboratory to detect subtle male factor problems. The results may be helpful for planning IVF, intracytoplasmic sperm injection, or varicocele surgery for infertile men. PMID- 10690767 TI - Semen analysis at the turn of the century: an evaluation of potential uses of new sperm function assays. AB - Semen analysis is a critical assay in the evaluation of infertility and may yield critical information regarding the etiology and prognosis of many types of reduced male fertility. However, basic semen analysis does not directly measure sperm fertilizing capacity, or many of the biochemical events both prior to and subsequent to fertilization. In the last two decades numerous assays of sperm function have been developed. These assays can be classified as: 1) Assays of general biochemistry and ultrastructure, 2) Assays of zona binding and oocyte penetration, and 3) Assays of postpenetration events. Sperm function assays not only allow an accurate diagnosis of many infertilities not diagnosed by the semen analysis, but can also lead to improved treatment modalities. In this review, basic semen analysis and many sperm function assays are briefly reviewed. Novel uses of sperm function are demonstrated in brief case studies. PMID- 10690768 TI - Mechanism of ejection during ejaculation: identification of a urethrocavernosus reflex. AB - The ejaculatory mechanism involves 2 reflexes: the "glans-vasal," which seems to bring the semen to the posterior urethra (emission phase of ejaculation), and the "urethromuscular" which ejects it to the exterior (ejection phase). This study investigated the mechanism of bulbocavernosus muscle (BCM) contraction, once the seminal fluid reaches the bulbous urethra. The study included 14 healthy male volunteers (mean age 37 +/- 10.2 SD years). To test the response of the BCM to urethral distension, a 10F balloon-tipped catheter was introduced into the prostatic urethra and filled with saline in increments of 0.25 mL: a needle electrode recorded the response. The balloon was then withdrawn to lie in the membranous. bulbous, and pendulous urethra and the test was repeated at each site. The latency of the muscle response was calculated. The BCM response to each of the anesthetized bulbous urethra and anesthetized BCM was recorded. Distension of the prostatic, membraneous, or pendulous urethra effected no BCM EMG response. Bulbous urethral distension with 0.25 mL of saline also produced no muscle response, whereas distension with 0.5 mL and up to 1.5 mL caused increased EMG activity of the BCM. The muscle response augmented with the increase of the distending volume. The mean latency was 10 +/- 1.3 ms and showed no significant change (p > .05) with the different distending volumes. Neither the anesthetized bulbous urethra nor the anesthetized BCM responded to bulbous urethral distension. The BCM contraction upon distension of the bulbous urethra is probably reflex and mediated through the urethrocavernosus reflex. Small-volume distension did not effect BCM contraction. The latter presumably propels the semen from the posterior to the pendulous urethra. It is suggested that the urethrocavernosus reflex be included in current andrologic investigations for patients with ejaculatory disorders. PMID- 10690769 TI - Porcine circoviruses: a review. AB - Porcine circoviruses (PCV) are small nonenveloped DNA viruses containing a unique single-stranded circular genome. Previously, no recognized link was found between PCV infection of pigs and disease, and PCV was considered a nonpathogenic agent. Over the last 5 years, a "novel" PCV, designated PCV2, has been associated with various disease syndromes in pigs, primarily postweaning multisystemic wasting syndrome (PMWS). Pigs with PMWS have a variety of clinical signs, including debility, dyspnea, palpable lymphadenopathy, diarrhea, and pallor or icterus. Lesions associated with the presence of PCV2 in a variety of cell types include lymphohistiocytic to granulomatous interstitial pneumonia, hepatitis, nephritis, myocarditis, enteritis, and pancreatitis. The lesions of PMWS have been reproduced experimentally after inoculation of piglets with PCV2 cell culture isolates, although the full expression of the disease syndrome may require the presence of other agents such as porcine parvovirus or porcine reproductive and respiratory syndrome (PRRS) virus. Recent reports have linked PCV2 to other disorders in pigs, ranging from abortion and reproductive failure to "atypical" PRRS. Available data indicate high seroprevalence of antibodies to PCV2 worldwide. The diagnosis of PCV2-associated disease is based on the direct demonstration of PCV2 antigens or nucleic acid in affected tissues. PCV2 is now regarded as an important emerging pathogen. Although vertical transmission has been documented, the epidemiology of PCV2 infections is poorly understood, as is the role of the immune response in controlling or augmenting disease. PMID- 10690770 TI - Gas chromatography/mass spectrometry identification and quantification of isazophos in a famphur pour-on and in bovine tissues after a toxic exposure. AB - A sample identified as "Warbex pour-on," expected to contain 13.2% famphur, and bovine tissue samples from 2 heifers that died after exhibiting signs of organophosphate intoxication were analyzed by gas chromatography/mass spectrometry (GC/MS). A product formulation problem was suspected because brain cholinesterase activities were depressed in both animals. Electron impact (EI) GC/MS of the pour-on revealed 9.7% famphur and an unidentified peak with approximately 76% of the peak area of the famphur. The unidentified peak showed a molecular ion at m/z 313, with a single Cl isotope cluster. Methane chemical ionization (MeCI) MS confirmed the molecular weight at 313 (1 Cl). A search on the molecular formula C9H17N3O3PSCl yielded a single match, isazophos. EI and MeCI GC/MS of reference isazophos confirmed the identity of the suspect peak. The concentration of isazophos in the pour-on was determined to be 6.0%. Famphur and isazophos were identified by their EI spectra and GC retention times in extracts of liver and brain from the 2 deceased animals. A GC/MS procedure utilizing selected ion monitoring (SIM) was developed for quantification of isazophos in liver, kidney, muscle, and fat of additional affected animals sacrificed at various times after exposure. Isazophos remained in animal tissues for as long as 94 days after topical exposure. Isazophos was present in fetal liver 70 days after exposure of the dam. High levels (6-3,500 ppm) of isazophos and famphur remained on the skin at 39 days postexposure. PMID- 10690771 TI - Coinfection by porcine circoviruses and porcine parvovirus in pigs with naturally acquired postweaning multisystemic wasting syndrome. AB - Postweaning multisystemic wasting syndrome (PMWS) is an emerging disease in swine. Recently, the disease has been reproduced with inocula containing a newly described porcine circovirus (PCV), designated PCV 2, and porcine parvovirus (PPV). In order to determine if these viruses interact in naturally acquired PMWS, affected tissues from field cases were examined by immunohistochemistry (IHC) and polymerase chain reaction (PCR) for PCV 2 and PPV, as well as by PCR for the other recognized porcine circovirus, PCV 1. Porcine circovirus 2 was detected by PCR or IHC in affected fixed or frozen tissues from 69 of 69 cases of PMWS collected over 3 years from 25 farms. Porcine parvovirus was detected in 12 of the same cases, and PCV 1 was detected in 9 of 69; however, an apparent decrease was found in the sensitivity of the PCRs used to detect the latter 2 viruses when fixed tissue from the same cases were compared with the use of frozen tissues. Porcine circovirus 2 was not detected by PCR in affected tissues from 16 age-matched pigs that had Streptococcus suis-associated disease. Electron microscopic examination of plasma pooled from 15 pigs with PMWS revealed the presence of PCV and PPV, whereas these viruses were not observed in pooled plasma from 5 age-matched clinically normal pigs. These results confirm and extend previous findings documenting a consistent association of PCV 2 with PMWS. As well, infection by PPV or PCV 1 or both may be an important cofactor in the pathogenesis of some, but apparently not all, cases of PMWS. PMID- 10690772 TI - Improvement of western blot test specificity for detecting equine serum antibodies to Sarcocystis neurona. AB - Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by the apicomplexan protozoan parasite Sarcocystis neurona. The purposes of this study were to develop the most stringent criteria possible for a positive test result, to estimate the sensitivity and specificity of the EPM Western blot antibody test, and to assess the ability of bovine antibodies to Sarcocystis cruzi to act as a blocking agent to minimize false-positive results in the western blot test for S. neurona. Sarcocystis neurona merozoites harvested from equine dermal cell culture were heat denatured, and the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in a 12 20% linear gradient gel. Separated proteins were electrophoretically transferred to polyvinylidene fluoride membranes and blocked in 1% bovine serum albumin and 0.5% Tween-Tris-buffered saline. Serum samples from 6 horses with S. neurona infections (confirmed by culture from neural tissue) and 57 horses without infections (horses from the Eastern Hemisphere, where S. neurona does not exist) were tested by Western blot. Horses from both groups had reactivity to the 62-, 30-, 16-, 13-, 11-, 10.5-, and 10-kD bands. Testing was repeated with another step. Blots were treated with bovine S. cruzi antibodies prior to loading the equine samples. After this modification of the Western blot test, positive infection status was significantly associated with reactivity to the 30- and 16 kD bands (P<0.001, Fisher's exact test). The S. cruzi antibody-blocked Western blot had a sample sensitivity of 100% and sample specificity of 98%. It is concluded that the specificity of the Western blot test is improved by blocking proteins not specific to S. neurona and using reactivity to the 30- and 16-kD bands as the criterion for a positive test. PMID- 10690773 TI - Bovine viral diarrhea virus cytopathic and noncytopathic biotypes and type 1 and 2 genotypes in diagnostic laboratory accessions: clinical and necropsy samples from cattle. AB - One hundred three bovine samples submitted to the Oklahoma Animal Disease Diagnostic Laboratory (OADDL) that were positive for bovine viral diarrhea virus (BVDV) were typed by a nested reverse transcription-polymerase chain reaction for BVDV genotypes. These BVDV samples included supernatants from virus isolation (79), serums (17), and buffy coats (7). The biotype, cytopathic (CP) or noncytopathic (NCP), was determined by cell culture virus isolation. Twenty-eight of 103 samples were submitted for herd screening for BVDV, 32 from OADDL necropsy cases, and 43 from live cattle with varied clinical conditions. Two samples contained 2 bands indicating presence of both BVDV types 1 and 2. Of the 105 BVDV samples, 26 were type 1 CP strains (24.8%), 38 were type 1 NCP strains (36.2%), 10 were type 2 CP strains (9.5%), and 31 were type 2 NCP strains (29.5%). From the 105 BVDV isolates, NCP biotypes were isolated more frequently (69, 65.7%) than CP biotypes (36, 34.3%), and type 1 genotypes were more frequently isolated (64, 61.00%) than type 2 genotypes (41, 39.0%). The NCP strains were more common than CP in herd screening samples. Cattle with respiratory disease history at time of sampling had more NCP than CP biotypes and more type 1 than type 2 genotypes. Of the necropsy cases, more were type 1 than type 2 genotypes for the respiratory cases with fibrinous pneumonia, more were type 1 than type 2 genotypes in cattle with enteritis/colitis without systemic lesions, and more were CP than NCP biotypes in cattle with enteritis/colitis with systemic lesions. No CP biotype was isolated from serum samples. PMID- 10690774 TI - Common variable immunodeficiency in miniature dachshunds affected with Pneumonocystis carinii pneumonia. AB - Seven miniature dachshunds, all under the age of 1 year, were presented with polypnea, tachypnea, and exercise intolerance as a result of Pneumocystis carinii pneumonia, which was diagnosed on transtracheal aspirate cytology. In all of the dogs, historical and clinical signs were suggestive of immune incompetence. Immunological studies undertaken were leukogram parameters, serum immunoglobulin fraction quantification, lymphocyte transformation assay. CD3 and CD79a lymphocyte markers on lymphoid tissue, and anti-canine immunoglobulin G immunoperoxidase staining. The immunological studies showed hypogammaglobulinemia, deficiency of serum immunoglobulins A, G, and M, decreased lymphocyte transformation response to phytohemagglutinin and pokeweed mitogens and absence of B lymphocytes with presence of T lymphocytes in the lymphoid tissue stained with CD3 and CD79a lymphocyte markers. The preceding findings suggest that P. carinii pneumonia occurring in the miniature dachshund is a result of both a T- and B-cell deficiency. This presentation is not the classic primary severe combined immunodeficiency syndrome but rather combined variable immunodeficiency, which has been well documented in humans but never in the dog. PMID- 10690775 TI - Procedurally similar competitive immunoassay systems for the serodiagnosis of Babesia equi, Babesia caballi, Trypanosoma equiperdum, and Burkholderia mallei infection in horses. AB - Procedurally similar competitive enzyme-linked immunoassay (cELISA) methods were developed for the serodiagnosis of Babesia equi and Babesia caballi (piroplasmosis), Trypanosoma equiperdum (dourine), and Burkholderia mallei (glanders) infections in horses. Apparent test specificities for the B. equi, B. caballi, T. equiperdum, and B. mallei cELISAs were 99.2%, 99.5%, 98.9%, and 98.9%, respectively. Concordances and kappa values between the complement fixation (CF) and the cELISA procedures for the serodiagnosis of B. equi, B. caballi, T. equiperdum, and B. mallei infections in experimentally exposed horses were 76% and 0.55, 89% and 0.78, 97% and 0.95, and 70% and 0.44, respectively. The cELISA method may be a technically more reproducible, objective, and convenient approach for piroplasmosis, dourine, and glanders serodiagnosis in qualifying animals for international movement and disease eradication programs than the CF systems currently in use. Use of the cELISA method also obviated the problems associated with testing hemolyzed or anticomplementary sera. PMID- 10690776 TI - Correlation of two nonradioactive immunoassays to a radioimmunoassay technique for thyroxine measurement in equine serum. AB - The purpose of this study was to compare 2 different nonradioactive assay methods with a conventional radioimmunoassay (RIA) measuring the concentration of serum thyroxine (T4) in horses. Serum was obtained from 85 adult standardbred horses. The T4 concentration of each sample was analyzed by RIA, chemiluminescent enzyme immunoassay (CEI), and homogeneous enzyme immunoassay (HEI). The correlation between the HEI method and RIA method was significantly greater (r = 0.89) than the correlation between the CEI and the reference method (r = 0.53). In addition, the precision of the HEI method was significantly greater than the CEI method; within-run percentage coefficients of variation were 4.5% and 15.9%, respectively, at mean T4 concentrations of 19-20 nmol/liter. On the basis of these findings, the HEI method was evaluated further. Both between-run precision and linearity were deemed adequate upon dilution by the HEI method. In addition, recovery of L-thyroxine added to equine serum was linear over 6 concentrations tested and averaged 79.6% with a manufacturer recommended data correction factor. An in-house correction factor was calculated by linear regression analysis of the RIA and HEI results from the original equine serum samples. Use of this correction factor improved the average recovery to 94.2% while maintaining excellent linearity (r2 = 0.9978). Although both nonradioactive methods of T4 analysis could likely substitute for the RIA reference method, the HEI method had the highest correlation and precision. The HEI technique also yielded adequately accurate results after correction of the data with an appropriate correction factor. Individually derived in-house correction factors may improve the accuracy of the HEI method to a greater extent than manufacturer suggested correction factors. PMID- 10690777 TI - Detection and differentiation of Mycobacterium avium and Mycobacterium genavense by polymerase chain reaction and restriction enzyme digestion analysis. PMID- 10690778 TI - Immunohistochemical detection of Mycobacterium paratuberculosis in formalin fixed, paraffin-embedded bovine tissue sections. PMID- 10690779 TI - Influence of storage and temperature on endospore and enterotoxin production by Clostridium perfringens in dogs. PMID- 10690780 TI - Isolation of avian paramyxovirus serotype 3 from domestic fowl in Israel: close antigenic relationship with the psittacine strain of avian paramyxovirus serotype 3. PMID- 10690781 TI - Pituitary abscess in young calves associated with the use of a controlled suckling device. PMID- 10690782 TI - Extradural myelolipoma in a dog. PMID- 10690783 TI - Diagnostic performance of a reverse transcription-polymerase chain reaction test for porcine reproductive and respiratory syndrome virus. PMID- 10690785 TI - Isolation of Jamestown Canyon virus (California virus group) from vesicular lesions of a horse. PMID- 10690784 TI - Companion animals as reservoirs of eaeA+ Escherichia coli. PMID- 10690786 TI - Improved plating media for the detection of Salmonella species with typical and atypical hydrogen sulfide production. AB - The Salmonella detection ability of 2 surfactant-supplemental media, xylose lysine-tergitol (Nia-proof) 4 (XLT4) and Miller-Mallinson (MM) agar, was compared against that of several commonly used plating media. XLT4 and MM appeared to be the most efficient in detecting Salmonella in meat products and food animal environments. MM was superior to XLT4 in detecting those increasingly more prevalent strains of Salmonella possessing weak to ultraweak H2S production characteristics. PMID- 10690787 TI - Canine distemper virus infection in binturongs (Arctictis binturong). PMID- 10690788 TI - Intrapelvic hemangiosarcoma in a horse. PMID- 10690789 TI - Pneumococcal vaccines for the world. PMID- 10690790 TI - A brief history of pneumococcal vaccines. AB - Attempts to control pneumococcal infection by vaccination, undertaken initially in 1911, have gone through 3 phases during the subsequent 8 decades. Initially, vaccines of killed pneumococcal cells prepared in a variety of ways were used in epidemic settings with inconclusive results, although administered to approximately 1 million recipients. The discovery that adults injected with small amounts of purified capsular polysaccharide developed antibodies to the homologous capsular type led to the trial of a tetravalent vaccine that showed conclusively its ability to prevent infection by the types represented in it. With the advent of penicillin and other effective antipneumococcal drugs, interest in prophylaxis waned. Interest in vaccination was revived only after demonstration that some segments of the population remained at high risk of death if infected and after the emergence of multidrug-resistant pneumococci. Infants and young children, among whom the incidence of pneumococcal infection is high, respond poorly to purified bacterial polysaccharides but develop satisfactory responses to bacterial polysaccharides when these are linked chemically to a protein. The early results of trials with such polysaccharide protein conjugate vaccines give promise that control of a significant portion of pneumococcal infection in the paediatric population will soon be feasible. PMID- 10690791 TI - Epidemiology of pneumococcal infections in the elderly. AB - The risk of invasive Streptococcus pneumoniae infection (primarily bacteraemia and meningitis) is greatest among the very young and the very old. Persons in certain racial groups, including African-Americans, American Indians, Native Alaskans and Australian Aborigines, are also at increased risk of disease. Other factors that appear to increase the risk of pneumococcal infection are lower socioeconomic status, recent infection with influenza and possibly other viral respiratory tract infections, chronic medical conditions, and immunosuppressive medications. Reported annual incidences of invasive pneumococcal disease among persons aged > or = 65 years in North America and Europe range from 25 to 90 cases/100,000 persons. In the US and Canada, these rates represent between 15,000 and 30,000 cases annually among the elderly. Mortality caused by pneumococcal infections is highest among the elderly, with nearly 1 in 5 cases resulting in death. Worldwide, S. pneumoniae is the leading cause of community-acquired pneumonia requiring hospitalisation. The high fatality rates, as well as recent outbreaks of pneumococcal infection among unvaccinated nursing home residents and the emergence of drug-resistant pneumococcal strains, highlight the importance of preventing invasive infection by vaccination. PMID- 10690792 TI - Pneumococcal vaccination for older adults: the first 20 years. AB - During the 20 years following its licensure, pneumococcal vaccine has not been widely used. Although the vaccine was shown to be efficacious in South African gold miners, clinical trials of 'pneumonia vaccine' in older adults that have attempted to demonstrate vaccine efficacy in preventing pneumonia have been inconclusive. Retrospective studies have convincingly demonstrated the effectiveness of vaccination in preventing invasive pneumococcal disease, but these findings have only gradually gained acceptance, largely because some observers reject the findings of observational studies or fail to appreciate the importance of invasive disease. In the 1980s, pneumococcal vaccine was used only in the US, but other countries began vaccination in the mid-1990s, in part due to a better understanding of the disease and the vaccine, but also because of concern about antimicrobial resistance. With greater understanding of the global importance of pneumococcal disease and the promise of conjugate and protein vaccines, during the next 20 years pneumococcal vaccines will become the most important vaccines for adults and children worldwide. PMID- 10690793 TI - Assessing the potential cost effectiveness of pneumococcal vaccines in the US: methodological issues and current evidence. AB - Pneumococcal disease imposes a notable burden on society, particularly in the elderly and those at high risk of complications. Preventive strategies, especially vaccines, are possibly the best way to minimise such a burden. We report on the conduct and results of a preliminary exploratory review of the economics of pneumococcal vaccines in the elderly population in the US. After extensive electronic and manual searches, we identified 5 economic evaluations that fulfilled our study criteria. From these we extracted key economic variables and assessed the quality of the studies against the criteria in the checklist for authors and peer reviewers of economic submissions to the British Medical Journal. We found variation of quality of study design such as a lack of clarity in the treatment of indirect costs and a failure to present the data on resource use and costs separately. We carried out supplementary searches to assess the quality of the epidemiological and efficacy evidence upon which the economic models were based and found contradictory evidence of effects of the vaccines, which included the results of 2 meta-analyses. One of these meta-analyses reported that retrospective studies, especially case-control studies, tended to underestimate the protective efficacy of the vaccine by as much as 20%. We believe that a well resourced Cochrane review of the clinical evidence of the effects of the vaccines should be carried out before any further economic studies. No more economic modelling should take place before such a review is undertaken. PMID- 10690795 TI - Safety of simultaneous pneumococcal and influenza vaccination in elderly patients in Brazil. PMID- 10690794 TI - Pneumococcal vaccine in the elderly: the Norwegian experience. AB - Pneumococcal infections have increased during the last 10 to 15 years in Norway. Their incidence is now about 20 per 100,000 population for all age groups, but is 2 to 3 times higher among the elderly. In 1996, the Advisory Board of Infectious Disease Control, National Institute of Public Health, Norway, recommended that pneumococcal polysaccharide vaccine should be administered to all individuals aged > or = 65 years. This recommendation has led to an increased use of pneumococcal vaccine, with a marked peak during the influenza vaccination season. PMID- 10690796 TI - Potential biochemical markers of uterine receptivity. AB - Implantation in humans is a complex process that involves embryo apposition and attachment to the maternal endometrial epithelium, traversing adjacent cells of the epithelial lining, and invasion into the endometrial stroma. These processes involve a variety of molecules which are not unique in themselves, but play unique roles in the process of implantation. The molecular dialogue that occurs between the implanting conceptus and the endometrium involves cell-cell and cell extracellular matrix interactions, mediated by lectins, integrins, matrix degrading enzymes and their inhibitors, prostaglandins, and a variety of growth factors, cytokines, and angiogenic peptides, their receptors and modulatory proteins. It is likely that each of these, when appropriately expressed or inhibited, contributes to endometrial receptivity or non-receptivity to an implanting conceptus. Currently, a working definition of a receptive versus a non receptive endometrium is incomplete. While histological normality of the endometrium does not necessarily imply functional normality, temporal and spatial expression of particular biochemical principles in the endometrium are highly suggestive of functional roles of these principles in implantation and endometrial receptivity. These potential markers of endometrial receptivity are discussed herein. It is envisioned that as regulation of these markers is elucidated, their expression may be manipulated to improve implantation rates and fertility or to limit implantation for successful contraception. PMID- 10690797 TI - MUC-1 glycosylation in endometrium: possible roles of the apical glycocalyx at implantation. AB - MUC-1 is a major epithelial apical surface glycoprotein in human endometrium. It has a large, extended and highly glycosylated ectodomain that contains keratan sulphate chains. MUC-1 is abundant at the luminal epithelial surface in the receptive phase, but keratan sulphate disappears at this time. MUC-1 has been shown experimentally to inhibit cell-cell interactions by steric hindrance of binding interactions mediated by receptors, including integrins and cadherins, so its high abundance at the time of implantation is unexpected. Here, various models for MUC-1 function in implantation are considered and its expression in different species compared. The possible evolutionary advantages of a maternal 'barrier' to implantation are discussed. PMID- 10690798 TI - Immunological aspects of implantation and implantation failure. AB - The human endometrium contains a significant proportion of leukocytes (8-35% of all cells), the absolute numbers and proportions varying during both the menstrual cycle and early in pregnancy. T cells, macrophages and a population of phenotypically unusual large granular lymphocytes (LGL) are commonly present, although B cells are absent. Relative T cell numbers decrease significantly in first trimester decidua, and hence are unlikely to play an important role in maintenance of human pregnancy, but T cells could be important in implantation where their relative numbers are greater. In addition to producing cytokines, local tissue macrophages may provide an immediate antigen non-specific host defence to infection. Most attention has, nevertheless, focused on a role for LGL in implantation and maintenance of pregnancy since, at the time of implantation, LGL comprise 70-80% of the total endometrial leukocyte population. Although endometrial LGL have been shown to express natural killer (NK) cell-type cytotoxicity against classical NK cell targets, such cytotoxicity against trophoblast is induced only after activation by interleukin (IL)-2. Selective expression of the unusual class I human leukocyte antigen (HLA) molecule, HLA-G, by extravillous cytotrophoblast may assist in protecting invasive cytotrophoblast from potential maternal NK cell attack, probably via interactions with killer inhibitory receptor molecules on LGL. Many cytokines have been demonstrated to be expressed at the maternal-fetal interface although, currently, in mice only two (IL-11 and leukaemia inhibitory factor) appear to be absolutely essential for successful pregnancy outcome. Immune effector cells and cytokines may also play a role in human pregnancy pathologies, such as recurrent early pregnancy loss. PMID- 10690799 TI - Cell-surface morphological events relevant to human implantation. AB - Morphological evidence on early stages of human implantation is limited to very few sporadic observations. The nature of implantation which requires the presence of both maternal and embryonic tissues, combined with the currently existing ethical constraints on human studies, appear to preclude generation of new data. However, research on relevant animal and in-vitro models as well as studies on human endometrium and in-vitro embryos, allow some indirect insights to this phenomenon. This review summarizes information on cell-surface morphological events relevant to implantation initiation, derived from scanning electron microscopy studies on the above systems. A central part of this article deals with the formation of epithelial cell projections known as pinopodes, for there is increasing evidence suggesting that these structures are closely associated with the development of endometrial receptivity for blastocyst implantation in humans. PMID- 10690800 TI - Blastocyst invasion and the stromal response in primates. AB - One of the most remarkable processes associated with the establishment of pregnancy in the primate is the process of decidualization. This transformation of a stromal fibroblast to a fully differentiated decidual cell is required for implantation and embryo survival in early pregnancy. Although the morphological and biochemical characteristics of the primate decidual cell have been extensively studied, the precise cellular, biochemical and molecular signals required for this transformation have yet to be elucidated. During decidualization, stromal cells first proliferate and then differentiate. Based on our extensive in-vivo and ongoing in-vitro studies, we have suggested that the process of decidualization in the baboon can be divided into two distinct phases. The initial proliferative phase is characterized by the expression of the cytoskeletal protein alpha smooth muscle actin (alphaSMA) in the stromal fibroblasts and is independent of the presence of the conceptus. The second phase of differentiation is characterized by the expression of insulin-like growth factor binding protein-1 (IGFBP-1) and the down-regulation of alphaSMA in the decidualized stromal fibroblast. The expression of IGFBP-1 is dependent on the presence of the conceptus in vivo and is regulated by hormones and cAMP in vitro. We have postulated that, during the initial phase of stromal cell differentiation, alphaSMA expression is regulated by the interaction between stromal cell integrins with the secreted extracellular matrix proteins (ECM). In response to pregnancy a trophoblast 'factor', mediated by cAMP signal transduction, induces IGFBP-1 expression in decidualizing stromal fibroblasts. This induction of IGFBP-1 is associated with the disappearance of alphaSMA and de novo protein synthesis. Our comparative studies suggest that the process of decidualization in the human and baboon involve similar mechanisms. However, the metabolic pathways required for decidualization in the two species appear to differ in their degree of sensitivity to external stimuli. This review focuses on the cellular events that may potentially regulate decidualization in the primate and its role in regulating trophoblast migration. PMID- 10690801 TI - Invasive cytotrophoblast apoptosis in pre-eclampsia. AB - Pre-eclampsia is a serious pregnancy complication diagnosed by signs of widespread maternal endothelial dysfunction. In normal pregnancy, a subpopulation of placental cytotrophoblast stem cells executes a differentiation programme that leads to invasion of the uterus and its vasculature. This process attaches the conceptus to the uterine wall and starts the flow of maternal blood to the placenta. In pre-eclampsia, cytotrophoblasts fail to differentiate along the invasive pathway. The functional consequences of this abnormality negatively affect interstitial and endovascular invasion, thereby compromising blood flow to the maternal-fetal interface. To determine whether abnormal differentiation and/or hypoxia leads to apoptosis of invasive cytotrophoblasts, we used the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling) method to label DNA strand breaks in tissue sections of the placenta and the uterine wall to which it attaches. Control samples (n = 9) showed little or no apoptosis in any location, but in samples from patients with pre-eclampsia, 15-50% of the cytotrophoblast subpopulation that invaded the uterine wall was labelled (8/9 samples). These same cells failed to stain for Bcl-2, a survival factor normally expressed by trophoblasts in both the placenta and the uterine wall. Our results show that pre-eclampsia is associated with widespread apoptosis of cytotrophoblasts that invade the uterus. The magnitude of programmed cell death in this population may account for the sudden onset of symptoms in some patients, as well as the associated coagulopathies. PMID- 10690802 TI - The relevance of the peritoneal fluid in endometriosis-associated infertility. AB - Endometriosis, one of the most prevalent gynaecological disorders, may affect fertility. Extensive research has been done in an attempt to understand the pathogenesis of endometriosis and its association with reproductive failure. It has been suggested that the disease affects almost any step of reproduction, but data are mostly controversial so it is difficult to draw clear conclusions from studies that have been done so far. Not only is peritoneal fluid in close proximity to endometriotic lesions, but it is also the environment in which early reproductive events take place. Studies on the peritoneal fluid in endometriosis have provided significant data towards an understanding of this disease. Immunological factors play a key role in determining the occurrence of endometriosis as well as its heterogenous symptoms. Since data also indicate that there are immunological differences between infertile and fertile women with endometriosis, recent studies have been designed to take these differences into consideration. This review will discuss the mechanisms by which endometriosis may affect fertility, and an emphasis will be placed on the relevance of the peritoneal fluid. PMID- 10690803 TI - Is the endometrium or oocyte/embryo affected in endometriosis? AB - One of the most puzzling problems of endometriosis is determining which mechanisms link this spectrum of conditions to infertility. There is conflicting evidence about the effect of endometriosis on the endometrium and on oocyte/embryo quality. Clinical studies reveal that implantation rates seem to be lower in women with endometriosis, while spontaneous abortion rates show variable results which are difficult to interpret due to the design of the studies. Biochemical markers (integrins and other cell adhesion molecules), morphological markers (pinopodes), apoptosis and ultrasound studies confirm that not only does the endometrium from women with endometriosis behave differently from the endometrium of women without endometriosis, but ectopic endometrium also behaves differently from eutopic endometrium. Data from oocyte donation programmes suggest that oocyte quality may be hampered in women with endometriosis. Recent reports have focused on the molecular mechanisms that may be altered, such as ovarian steroid production, or inadequate luteal function. In this review, we analyse the most recent literature dealing with the different mechanisms which affect the endometrium and oocyte/embryo quality and which thereby might cause infertility. PMID- 10690804 TI - Role of the IGF system in trophoblast invasion and pre-eclampsia. AB - Insulin-like growth factor-II (IGF-II) and IGF binding protein-1 (IGFBP-1) appear to play an important role in paracrine interactions at the maternal-fetal interface in human pregnancy. Patterns of expression of IGF-II and IGFBP-1 at the decidual-trophoblast interface suggest paracrine interactions occur between the IGF-II-expressing invading cytotrophoblast and maternal decidua-derived IGFBP-1. Autocrine/paracrine actions of trophoblast-derived IGF-II may be important in invasion, and for both trophoblast and decidual function. The actions of IGFBP-1 in binding IGF, and as an integrin ligand, suggest it may have multiple roles in the interactions between the invading trophoblast and the maternal decidua. Abundant decidual IGFBP-1 may interact with the IGF-II-expressing, protease secreting trophoblast to modulate invasion. In-vitro studies of trophoblast decidual cell interactions in invasion, and clinical observations in a gestational disorder with shallow placental invasion such as pre-eclampsia, have provided new insights into the possible role(s) of IGFBP-1 in trophoblast invasion. The precise mechanisms underlying IGF and IGFBP-1 action at the decidual-trophoblast interface remain to be elucidated. The potential predictive value of serum IGFBP-1 concentrations in pre-eclampsia also remains to be established. PMID- 10690805 TI - Pinopodes as markers of endometrial receptivity in clinical practice. AB - Clinical evidence indicates the existence in the human of a narrow window of uterine receptivity which opens during the mid-luteal phase. At the same time, formation of pinopodes on the apical membranes of the endometrial epithelial cells occurs. To develop a specific marker for receptivity in clinical practice, the kinetics of pinopode formation has been investigated through sequential endometrial biopsying in natural, hormone replacement and stimulated cycles. The results show that pinopodes last <48 h in all cycles, and the timing of their formation depends both on the hormonal treatment applied and on the patient's individual response. On average, pinopodes form earlier in stimulated cycles and later in hormone replacement cycles, compared with natural cycles. Pinopode expression is strongly correlated with implantation following embryo transfer and furthermore, detection of pinopodes in assessment cycles may be extremely useful for the assessment of receptivity on an individual basis to optimize embryo transfer, resulting in increased implantation rates. Finally, pinopodes seem to be correlated well with other cellular and molecular events occurring in the mid luteal endometrium. PMID- 10690806 TI - Ovarian stimulation and endometrial receptivity. AB - Endometrial receptivity is a limiting step in the success of in-vitro fertilization (IVF). To investigate this issue, we selected a specific population of high responder patients in whom implantation was impaired, even when good quality embryos were transferred. We present a series of studies showing that high oestradiol concentrations on the day of human chorionic gonadotrophin (HCG) administration are detrimental to uterine receptivity. In addition, we suggest clinical strategies to improve endometrial receptivity in high responder patients using a step-down regimen. PMID- 10690807 TI - Use of co-culture of human embryos on Vero cells to improve clinical implantation rate. AB - Co-culture of human embryos (n = 384 cycles) to the blastocyst stage using Vero cell monolayers was carried out between August 1995 and December 1997. A total of 2868 zygotes were co-cultured and 1027 embryos reached the blastocyst stage (blastocyst formation rate 35.8%). The blastocysts were frozen in 43.7% of patients. A mean of 1.8 blastocysts was transferred per patient and 95 pregnancies were obtained (pregnancy rate/cycle 24.7%). The blastocyst implantation rate was 23.6%. Miscarriage occurred in 15 patients (15.7%) and ectopic pregnancy in three (3.1%) patients. The multiple pregnancy rate was 32.6%. No differences were observed in the blastocyst rate between poor, normal or high response patients. Blastocyst formation was significantly lower when frozen donor spermatozoa were used. Significantly higher pregnancy rates per transfer and blastocyst implantation rates were attained when embryos were transferred on days 5 or 6 compared with day 7. No advantage was observed when co culture was used in first cycle IVF patients, in comparison with conventional day 2 replacements. The use of blastocysts for preimplantation genetic diagnosis (PGD) increases the diagnostic reliability and widens diagnostic possibilities. A total of 215 cycles with frozen-thawed co-cultured blastocysts were carried out, with a pregnancy rate of 22.7% per replacement. PMID- 10690808 TI - The mechanism of accommodation in primates. PMID- 10690809 TI - Underestimation of intraocular lens power for cataract surgery after myopic PRK. PMID- 10690810 TI - Augmentation of filtering blebs with perfluoropropane gas bubble. PMID- 10690811 TI - Augmentation of filtering blebs with perfluoropropane gas bubble. PMID- 10690812 TI - Sclerosing sweat duct carcinoma of the eyelid margin: unusual presentation of a rare tumor. PMID- 10690813 TI - Acute corneal necrosis after excimer laser keratectomy for hyperopia. PMID- 10690814 TI - Intraocular lens power calculation after corneal refractive surgery remains challenging. PMID- 10690815 TI - To my dismay PMID- 10690816 TI - The National Eye Institute's low vision education program: improving quality of life. PMID- 10690817 TI - The South Asian cataract management study: complications, vision outcomes, and corneal endothelial cell loss in a randomized multicenter clinical trial comparing intracapsular cataract extraction with and without anterior chamber intraocular lens implantation. AB - OBJECTIVE: To determine clinical outcomes of primary intracapsular cataract surgery with and without implantation of anterior chamber lenses. DESIGN: A multicenter randomized clinical trial. PARTICIPANTS: One thousand two hundred twenty-nine male and female patients 40-75 years of age with senile cataract. METHODS: Study patients were recruited from screening eye camps and outpatient clinics. Randomization to the two treatment groups was performed after screening for predetermined inclusion and exclusion criteria. Demographics, visual acuity, intraocular pressures, and corneal endothelial cell data were recorded before surgery and at 6 weeks, 12 months, and 24 months after surgery. Monitoring of the study was secured by a standardized image documentation procedure on all patients using the IMAGEnet digital imaging system. Analysis of corneal endothelial cell images was performed with the Cell Soft software (Topcon Corporation, Japan). MAIN OUTCOME MEASURES: Visual acuity and central corneal endothelial cell loss. RESULTS: The patients were randomized to intraocular lens (IOL; n = 616) and no IOL (n = 613) implantation. Surgical complications were reported in 177 (14.4%) patients (IOL = 14.8%; no IOL = 14.0%). The most frequent complication observed was vitreous loss which occurred in 10.3% of eyes (IOL = 11.2%; no IOL = 9.5%). At the final examination (2 years after surgery), 88% of the operated eyes had a best corrected vision of 6/18 or better (IOL = 88.8%; no IOL = 86.6%). Analysis of corneal endothelial cell data showed a small but significantly greater cell loss 6 weeks after surgery in eyes with IOL compared with those without IOL, but no overall difference was found between the treatment groups in the long term follow-up. CONCLUSIONS: The findings indicate that there is a rationale for the use of anterior chamber intraocular lenses in primary intracapsular cataract surgery. PMID- 10690818 TI - Inflammation after sclerocorneal versus clear corneal tunnel phacoemulsification. AB - OBJECTIVE: To compare the postoperative inflammation after phacoemulsification followed by intraocular lens (IOL) implantation by means of sclerocorneal versus clear corneal tunnel incision. DESIGN: Randomized controlled clinical trial. PARTICIPANTS: One hundred eyes of 100 patients were examined at a German University eye hospital. INTERVENTION: One hundred eyes with cataract necessitating phacoemulsification with posterior chamber IOL implantation were randomly assigned to receive a temporal sclerocorneal or clear corneal tunnel incision by a single surgeon. MAIN OUTCOME MEASURES: Preoperative and postoperative inflammation was evaluated by measurement of flare using laser flare photometry. Statistical inference was mainly based on nonparametric group comparisons by use of two sample Wilcoxon tests. RESULTS: Mean anterior chamber flare in the group with sclerocorneal tunnel increased from 7.5 photon counts/ms preoperatively to 19.6 at 6 hours postoperatively and decreased to 11.1 (day 1), 11.7 (day 2), 11.6 (day 3), and 9.2 (5 months) during the postoperative course. The mean flare in the clear corneal tunnel incision group increased from 7.7 preoperatively to 12.9 at 6 hours postoperatively and then decreased to 9.2 (day 1), 9.8 (day 2), 9.1 (day 3), and 9.2 (5 months). Individual postoperative flare changes were significantly lower in the clear corneal tunnel group at the day of surgery (P<0.0001), as well as at day 1 (P = 0.0011), day 2 (P = 0.0079), and day 3 (P = 0.0020). After 5 months, no statistically significant difference was found. CONCLUSIONS: After phacoemulsification and foldable IOL implantation, postoperative alteration in the blood-aqueous barrier was statistically significantly lower with the clear corneal tunnel incision group compared with the sclerocorneal incision group, in the first 3 days postoperatively. PMID- 10690819 TI - Subjective visual experience during phacoemulsification and intraocular lens implantation under topical anesthesia. AB - OBJECTIVE: The aim of this study was to investigate the subjective visual experience of patients during phacoemulsification and intraocular lens (IOL) implantation under topical anesthesia. DESIGN: Postoperative questionnaire survey. PARTICIPANTS: The study cohort consisted of 52 patients with cataracts. There were 18 male (34.6%) and 34 female (65.4%) patients. Their mean (+/- SD) age was 67.5 (+/-10.8) years. INTERVENTION: The patients underwent routine phacoemulsification and IOL implantation under topical anesthesia. MAIN OUTCOME MEASURES: The patients were interviewed on the same day after their operation regarding their visual experience in the operated eye during surgery. RESULTS: All patients (100%) reported that they could see at least some light during the surgery. Some patients reported they could also see one or more colors (50 patients, 96.2%), movements (32 patients, 61.5%), flashes (24 patients, 46.2%), the surgeon's fingers/hands (13 patients, 25%), instruments (12 patients, 23.1%), and/or the surgeon (4 patients, 7.7%). The colors seen included red (24 patients, 46.2%), yellow (23 patients, 44.2%), blue (12 patients, 23.1%), green (7 patients, 13.5%), and orange (6 patients, 11.5%). Eight patients (15.4%) saw the spectrum of colors similar to that of the rainbow. Twenty-four patients (46.2%) reported that the brightness of light changed during the course of the operation. Eight patients (15.4%) found their visual experience frightening. There was no statistically significant association between those who found the visual experience frightening and the sex or age of the patient, a history of cataract operation in the fellow eye, the type of visual sensation experienced, or the presence of coexisting ocular pathology. CONCLUSION: All patients undergoing phacoemulsification under topical anesthesia experience a variety of visual sensations that may be frightening in a small proportion of patients. PMID- 10690820 TI - Photorefractive keratectomy versus laser in situ keratomileusis: a control matched study. AB - OBJECTIVE: Photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) outcomes were compared at 1, 3, 6, and 12 months after surgery. DESIGN: Retrospective, nonrandomized, comparative study. PARTICIPANTS: One hundred seven LASIK-treated eyes (58 patients) and 107 PRK-treated eyes (91 patients) having myopia between -1 and -9.50 diopters (D). All LASIK-treated eyes were analyzed retrospectively and matched with PRK-treated eyes having sphere within +/-0.25 D, +/-1 D of cylinder, and +/-7 years of age. INTERVENTION: For PRK and LASIK, the refractive surgery was performed with the slit-scanning excimer laser Nidek EC 5000, (Nidek Co. Tokyo, Japan) MAIN OUTCOME MEASURES: Manifest refraction, best spectacle and uncorrected Snellen visual acuity, haze, halos, and keratometry were evaluated before surgery and up to 12 months after surgery. RESULTS: Seventy percent of eyes were evaluated at the 12-month postoperative exam. Of these eyes, 83% of LASIK cases and 86% of PRK cases had uncorrected visual acuities of 20/20 or better. Refractions within +/-0.5 D represented 78% of the LASIK eyes and 83% of the PRK eyes at that follow-up. Patients who underwent LASIK reported halos twice as often as patients who underwent PRK using a subjective scale. The odds ratio of high halos for LASIK versus PRK was 3.50 (95% confidence interval, 1.89 6.46; P<0.0001). At 1 month after surgery, 64% of the LASIK eyes were within +/ 0.50 D compared with 77% of the PRK eyes. No eye lost 2 Snellen lines of best corrected visual acuity at 6 or 12 months after surgery. Ten PRK eyes (9.3%) and three LASIK eyes (2.8%) were retreated. CONCLUSIONS: PRK and LASIK achieved equal refractive outcomes at all postoperative follow-ups, but LASIK patients were twice as likely to experience halos. PMID- 10690821 TI - Incidence of vitreoretinal pathologic conditions within 24 months after laser in situ keratomileusis. AB - OBJECTIVE: To report for the first time a case series of vitreoretinal pathologic conditions after laser in situ keratomileusis (LASIK) and to determine its incidence. DESIGN: Case series. PARTICIPANTS: Five refractive surgeons and 29,916 eyes that underwent surgical correction of ametropia (83.2% were myopic) ranging from -0.75 to -29.00 diopters (D; mean: -6.19 D) and from +1.00 to +6.00 D (mean: +3.23 D) participated in this retrospective study. MAIN OUTCOME MEASURES: Vitreoretinal complications after LASIK. RESULTS: The clinical findings of 20 eyes (17 patients) with LASIK-related vitreoretinal pathologic conditions are presented. Fourteen eyes experienced rhegmatogenous retinal detachments (RDs). Two eyes experienced corneoscleral perforations with the surgical microkeratome when a corneal flap was being performed (one experienced a vitreous hemorrhage and the other later experienced an RD). In four eyes, retinal tears without RDs were found. In one eye, a juxtafoveal choroidal neovascular membrane (CNVM) developed. Retinal tears were treated with argon laser retinopexy or cryotherapy. Corneoscleral perforations were sutured, and the RD was managed with vitrectomy. The remaining RDs were managed with vitrectomy, cryoretinopexy, scleral buckling, argon laser retinopexy, or pneumatic retinopexy techniques. The CNVM was surgically removed. The incidence of vitreoretinal pathologic conditions determined in our study was 0.06%. CONCLUSIONS: Serious complications after LASIK are infrequent. Vitreoretinal pathologic conditions, if managed promptly, will still result in good vision. It is very important to inform patients that LASIK only corrects the refractive aspect of myopia. Complications of the myopic eye will persist. PMID- 10690822 TI - A randomized trial of low-dose, topical mitomycin-C in the treatment of severe vernal keratoconjunctivitis. AB - OBJECTIVE: To evaluate the efficacy and safety of low-dose, topical mitomycin-C in patients with severe vernal keratoconjunctivitis. DESIGN: Placebo-controlled, double-masked, randomized clinical trial. PARTICIPANTS: Twenty-six patients with vernal keratoconjunctivitis refractory to combination of steroid and mast-cell stabilizer treatment. INTERVENTION: Patients were randomly assigned (2:1) to treatment with topical 0.01% mitomycin-C eye drops (n = 17) or placebo (n = 9) three times daily for 2 weeks. MAIN OUTCOME MEASURES: Symptoms (itching, tearing, photophobia, ropy mucous discharge, foreign body sensation) and signs (conjunctival hyperemia, epithelial punctate keratitis, Trantas' dots, limbal edema, and palpebral conjunctival giant papillae) of vernal keratoconjunctivitis recorded on the day of enrollment and at the end of the treatment period. RESULTS: There was a statistically significant decrease in ropy mucous discharge, photophobia, conjunctival hyperemia, and limbal edema in the mitomycin-C treated group compared with the placebo group at the end of the 2-week treatment period. In addition, none of the 17 treated patients, but all 9 of the placebo patients, required medication during the 4-week posttreatment follow-up period. No adverse effects of treatment with mitomycin-C were observed. CONCLUSIONS: Short-term, low dose, topical mitomycin-C may be considered in the acute exacerbation periods of patients with severe vernal keratoconjunctivitis refractory to conventional treatment. PMID- 10690823 TI - Limbal-conjunctival autograft transplantation for the treatment of recurrent pterygium. AB - OBJECTIVE: Different investigators have recently emphasized the importance of the limbus and its stem cells in the pathogenesis of the pterygium. In this article we examine the usefulness of limbal-conjunctival autograft transplantation for the treatment of advanced recurrent pterygium. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Seven patients with advanced recurrent pterygium. All had previously been treated a minimum of two times by simple excision (two of them with intraoperative mitomycin C). INTERVENTION: Limbal-conjunctival autograft transplantation after pterygium excision was performed in all cases. MAIN OUTCOME MEASURES: Pterygium recurrences and complications with a minimal follow-up period of 14 months. RESULTS: There were no recurrences of pterygial growth beyond the limbal edge. In addition, no significant complications were noted. Only one case of limited pseudopterygium in the donor site and one case of graft retraction were recorded. No further surgical interventions were needed in any case. CONCLUSIONS: Limbal-conjunctival autograft transplantation is a promising technique for the treatment of advanced recurrent pterygium. PMID- 10690824 TI - Conjunctival vasculature in the assessment of anemia. AB - OBJECTIVE: To assess the correlation of the bulbar conjunctival blood column (BCBC) with anemia. DESIGN: A prospective, randomized, masked, two observer case series. PARTICIPANTS: Inpatients on hospital wards; outpatients in both the Hematology-Oncology and Ophthalmology Clinics. METHODS: Observations of the palpebral conjunctival hue (PCH) and BCBC by two observers masked to the patient's diagnosis, laboratory test results, and other's observations. The PCH and BCBC were correlated by slit-lamp examination with serum hemoglobin values. Different threshold levels for anemia were defined as hemoglobin <10, <11, and <12 mg/dl. MAIN OUTCOME MEASURES: The parameters included determination of (1) the conjunctival hue, assessed as pink or pale and (2) the bulbar conjunctival blood column, assessed as full (normal), granular, or discontinuous. These data were compared against the patient's hemoglobin level. RESULTS: Mean hemoglobin was 11.0+/-2.2 mg/dl. Sensitivity of the BCBC and PCH for anemia was 83%-94% and 38%, respectively, regardless of the definition of anemia. Specificity of BCBC improved with increasing hemoglobin threshold levels for anemia: 56% (hemoglobin <10 mg/dl) to 73% (hemoglobin <12 mg/dl); specificity for PCH ranged from 82% to 94%. The BCBC was significantly (P<0.03) associated with anemia for hemoglobin <11 mg/dl for both observers (logistic regression, Spearman correlation). There was a significant (P<0.05) association of PCH with anemia only for hemoglobin <10 mg/dl with logistic regression (one observer only) and with Spearman correlation (both observers). CONCLUSIONS: The BCBC is significantly associated with anemia, with higher sensitivity and only slightly less specificity than PCH. PMID- 10690825 TI - Oral acyclovir for the management of herpes simplex virus keratitis in children. AB - PURPOSE: To evaluate the use of oral acyclovir in pediatric patients with herpes simplex virus (HSV) keratitis. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Seven pediatric patients seen at the University of Minnesota Hospitals and Clinics with herpes simplex virus (HSV) infectious epithelial keratitis between January 1992 and October 1998. Patient ages ranged from 6 weeks to 5 years at time of presentation with a median of 1.7 and mean of 1.9 years. INTERVENTION: All patients received oral acyclovir; six of seven patients also received topical antiviral medications. Three of seven patients had topical antiviral therapy fail before being placed on oral acyclovir, and the remaining four patients were placed on oral acyclovir primarily. RESULTS: All patients showed resolution of HSV infectious epithelial keratitis. Three patients have been maintained on prophylactic dosage of oral acyclovir because of recurrent disease or because they have been chronically treated with topical corticosteroids for immune stromal keratitis. All patients tolerated acyclovir well, and there were no adverse reactions. CONCLUSIONS: Oral acyclovir is useful in treating HSV infectious epithelial keratitis in pediatric patients. It is beneficial in treating infectious epithelial keratitis and prophylactically either while treating with topical corticosteroids for immune stromal keratitis or for preventing recurrent infectious epithelial keratitis. PMID- 10690826 TI - Comparison of the efficacy of betaxolol-brinzolamide and timolol-dorzolamide as suppressors of aqueous humor flow in human subjects. AB - OBJECTIVE: To compare the efficacy of combinations of betaxolol-brinzolamide and timolol-dorzolamide as suppressors of aqueous humor flow and ocular hypotensive agents. DESIGN: Placebo-controlled, masked comparison of the two drug combinations. PARTICIPANTS: Twenty-five normal human volunteers with the fellow eye serving as control. METHODS OR TESTING: Fluorophotometric measurement of aqueous humor flow and pneumatonometric measurement of intraocular pressure. MAIN OUTCOME MEASURES: Aqueous humor flow and intraocular pressure. RESULTS: The betaxolol-brinzolamide combination lowered aqueous flow 39% to 44%, and the timololdorzolamide combination lowered aqueous flow 51%. The betaxolol brinzolamide combination lowered intraocular pressure 14% to 19%, and the timolol dorzolamide combination lowered it 18% to 24%. CONCLUSIONS: Both drug combinations were effective; the timolol-dorzolamide combination appeared to be the more effective of the two after short-term exposure (24 hours). PMID- 10690827 TI - Frequency doubling technique in patients with ocular hypertension and glaucoma: correlation with octopus perimeter indices. AB - PURPOSE: To ascertain whether frequency doubling technique (FDT) (Welch-Allyn, Skaneateles, NY; Zeiss-Humphrey, San Leandro, CA) indices provide results comparable with those of standard Octopus threshold perimeters (Interzeag AG, CH 8952 Schlieren, CH) in patients with glaucoma and in patients suspected of having ocular hypertension, glaucoma, or both. DESIGN: A comparative, consecutive, case series. PARTICIPANTS: Thirty-nine glaucomatous patients and 41 patients with ocular hypertension or suspected glaucoma were recruited consecutively. METHODS: The visual field of the study participants were assessed by FDT program C-20 full threshold and Octopus program dG1X. Only one eye of each participant was selected randomly. Pearson's r correlation coefficient was calculated among the FDT and Octopus indices. MAIN OUTCOME MEASURES: Using Octopus perimeter, mean defect (MD), mean sensitivity (MS), loss variance (LV), and corrected loss variance (CLV) were calculated and used for correlation. For the FDT, mean deviation (FDT MD) and pattern standard deviation (FDT-PSD) were calculated and used for correlation. Also, the time required to perform the visual field test was considered. RESULTS: In the entire population, a statistically significant correlation (Pearson's r, P<0.001) was found between FDT-MD and both MS (0.77) and MD (-0.80) and between FDT-PSD and both LV (0.50) and CLV (0.45). When the glaucoma group was considered alone, similar significant correlation was found between the indices. In the suspected ocular hypertension and glaucoma suspect group, no significant correlation was found. A significant (P<0.001) difference was found between FDT and Octopus for the time needed to perform the visual field test. CONCLUSIONS: This new technique could be used both to screen populations and to observe glaucomatous visual field progression in early and moderate stages. The FDT is a faster way to analyze the visual field and captures threshold values for each point, but it is important to remember that this is a new technique and its limits are still unknown. PMID- 10690828 TI - Outcome of trabeculectomy with mitomycin-C in the iridocorneal endothelial syndrome. AB - PURPOSE: Eyes with iridocorneal endothelial (ICE) syndrome have a high risk of failure in glaucoma filtering surgery failing. We investigated the efficacy of trabeculectomy with intraoperative mitomycin-C application in these patients. DESIGN: Retrospective nonrandomized comparative trial with historical controls. PARTICIPANTS AND CONTROLS: Ten patients with unilateral iridocorneal endothelial (ICE) syndrome were reviewed. Their intraocular pressures could not be controlled medically. In five eyes, this was the primary surgery performed. Five of the patients had undergone prior intraocular pressure-(IOP) lowering surgery that had failed at the time enrolled. Results were compared with previously published case series of similar patients treated with trabeculectomy alone or trabeculectomy and subconjunctival 5-fluorouracil injections. INTERVENTION: Intervention consisted of trabeculectomy with a limbus-based conjunctival flap and mitomycin-C application. The dosage of mitomycin-C was 0.4 mg/ml for 1 to 4 minutes (mean, 1.9 min). MAIN OUTCOME MEASURES: Adequate control of IOP (without medication lower than 21 mm Hg). RESULTS: In eight eyes the IOP remained well controlled (mean IOP, 12.1 mm Hg) over the entire length of available of follow-up (mean, 14.9 months). Two eyes required implantation of an aqueous tube shunt at 4 and 11 months, respectively, after trabeculectomy with mitomycin-C. One eye experienced visual loss of 3 Snellen lines because of hypotony maculopathy. CONCLUSIONS: Trabeculectomy with mitomycin-C application offers a reasonable intermediate-term success rate in ICE patients, who are otherwise at high risk for failure of filtering surgery. PMID- 10690829 TI - Simultaneous subconjunctival and subscleral mitomycin-C application in trabeculectomy. AB - OBJECTIVE: To establish the efficacy and safety of simultaneous subconjunctival and subscleral application of mitomycin-C in trabeculectomy. DESIGN: A prospective, randomized study. PARTICIPANTS: Sixty-eight patients (68 eyes) with refractory glaucomas were included in the study. INTERVENTION: Eyes were randomly assigned to receive intraoperative mitomycin-C (0.3 mg/ml) applied under the conjunctival flap (group 1), scleral flap (group 2), or under both flaps (group 3). MAIN OUTCOME MEASURES: Mean intraocular pressure (IOP), postoperative medications, visual acuity, filtering bleb appearance, and complications. RESULTS: There was a significant difference in IOP at 6, 9, and 12 months after surgery among the three groups (P = 0.021, 0.026, and 0.033, respectively ANOVA). At 12 months, the mean IOP in group 3 was 9.8+/-3.7 mm Hg compared with 13.4+/ 5.5 mm Hg in group 2. (P = 0.015) and 12.4+/-4.4 mm Hg in group 1 (P = 0.039). Success rate (21 mm Hg or less), number of antiglaucoma medications, and complications showed no statistical significant difference between the three groups at each postoperative visit. CONCLUSIONS: Mitomycin-C applied under the scleral flap may have an additional beneficial effect when combined with simultaneous subconjunctival exposure. PMID- 10690830 TI - Determinants of glaucoma awareness in a general eye clinic. AB - PURPOSE: Heightened public awareness about glaucoma may increase the chance of identifying undetected cases. To ascertain determinants of glaucoma awareness, we surveyed a population visiting a general eye clinic. DESIGN: Cross-sectional study. PARTICIPANTS: 1197 general eye clinic patients and their companions. METHODS: We designed and administered a questionnaire about glaucoma to general eye clinic patients and their companions. We created multivariate logistic regression models to ascertain the effect of demographic and clinical features on the likelihood of being unaware of glaucoma. MAIN OUTCOME MEASURES: Adjusted odds ratio (OR) with 95% confidence intervals of survey attributes associated with self-perceived unfamiliarity with glaucoma. RESULTS: Glaucoma awareness overall (72%) approached that found in the subgroup self-reporting a diagnosis of glaucoma (80%). Survey attributes associated with an increased likelihood of being unaware of glaucoma were African American race (OR = 1.69 [1.28-2.20], Hispanic ethnicity (OR = 2.13 [1.46-3.02]), and less than a college education (OR = 1.67 [1.37-2.05]). Age was also a determinant of glaucoma awareness (for ages 50-64 years, OR = 0.60 [0.44-0.80] and for ages 65-79 years, OR = 0.56 [0.41 0.75] compared with ages less than 35 years). A self-report of glaucoma was not a determinant of glaucoma awareness (OR = 0.63 [0.33-1.17]), although there was a trend toward enhanced glaucoma awareness in this subgroup. Finally, respondents with a history of employment in the health field (OR = 0.63 [0.49-0.82]) myopia (OR = 0.68 [0.56-0.82]), glaucoma in a first-degree relative (OR = 0.68 [0.53 0.87]), and respondents who reported having a dilated eye examination (OR = 0.53 [0.42-0.66]) were less likely to be unaware of glaucoma than those who did not have these attributes. CONCLUSIONS: Although glaucoma awareness in this population was high, Hispanics, African Americans, and those with less than a college education were more likely to be unfamiliar with the disease. Interestingly, a self-report of having glaucoma was not a statistically significant determinant of glaucoma awareness. PMID- 10690831 TI - Outcomes of sequential tube shunts in complicated glaucoma. AB - OBJECTIVE: To evaluate intraocular pressure (IOP) control, change in visual acuity, and complications in eyes that have undergone a second glaucoma tube shunt procedure. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Twenty-two eyes of 22 patients that have undergone sequential tube implants for management of glaucoma. METHODS: Parameters analyzed included IOP, visual acuity, and number of hypotensive agent before each shunt procedure and at last follow-up visit. The overall IOP lowering effect attributable to each tube shunt was calculated. Any ocular complications after the second tube shunt were recorded. Success was defined as an IOP between 6 and 21 mm Hg and a 20% reduction in IOP from the second tube shunt procedure. Qualified successes met one of these two requirements at the last follow-up visit. Total failures did not meet any of the above criteria, required additional surgical intervention to lower IOP, or both. MAIN OUTCOME MEASURES: Intraocular pressure control, visual acuity preservation, and complications. RESULTS: At the last follow-up visit, the average percent reduction in IOP from both tube shunt procedures was 42+/-21%. The average percent IOP reduction from the second tube shunt was 33+/-17%. Eleven (50%) patients met the criteria for success, 8 (36.4%) patients were qualified successes, and 3 (13.6%) were failures. The median number of hypotensive agents decreased from two to one. Ten patients experienced new or worse pseudophakic bullous keratopathy after the second tube shunt, six of whom underwent penetrating keratoplasty. Thirteen (59%) patients maintained visual acuity within one line of their second tube shunt pre-operative Snellen visual acuity. Seven (32%) patients lost more than 2 lines, and one patient lost light perception. CONCLUSIONS: Although corneal morbidity is a common complication, a second tube shunt does not cause higher-than-expected rates of other complications associated with tube shunt surgery. Eyes that undergo a second tube shunt procedure can achieve pressure control, require fewer hypotensive agents, and may maintain stable visual acuity. PMID- 10690832 TI - Prevention of cellulitis after open lacrimal surgery: a prospective study of three methods. AB - OBJECTIVE: Soft-tissue cellulitis after open lacrimal surgery, almost exclusively caused by infection, is associated with a reduced surgical success rate, inconvenience for the patient, and may predispose to secondary hemorrhage. Although postoperative oral antibiotics have been shown to significantly reduce the infection rate, this investigation was designed to compare this regimen with two other methods for the prevention of postoperative infection. DESIGN: A prospective nonrandomized comparative trial of three methods for prevention of postoperative infection. PARTICIPANTS: Patients, recruited over a 16-month period from the Lacrimal Clinic at Moorfields Eye Hospital, who required primary open dacryocystorhinostomy. METHOD: Two hundred and sixty-five patients were assigned, on the basis of hospital number, to one of three groups: to receive either an intravenous broad-spectrum antibiotic immediately after induction of anesthesia (group 1), intraoperative lavage of the rhinostomy site just after suture of the anterior mucosal flaps (group 2), or a postoperative course of oral antibiotics (group 3). OUTCOME MEASURES: Cellulitis was considered to be present when there was marked tenderness along a swollen and erythematous incision line, evidence of frank pus from the suture line, or late postoperative hemorrhage occurred. Patients were reviewed within a week of surgery. RESULTS: Signs of infective cellulitis occurred in 1 of 127 (0.8%) cases in group 1, 7 of 41 (18%) cases in group 2, and 2 of 127 (1.5%) cases in group 3. Whereas the infection rate in group 2 was significantly greater than that in group 1 (P << 0.001) or group 3 (P<0.001), no significant difference was found between that in groups 1 and 3 (0.75>P>0.5; chi-square test with Yates' correction). CONCLUSION: Compared with intraoperative saline lavage, intraoperative or postoperative broad-spectrum antibiotics have comparable efficacy in the prevention of postoperative soft tissue cellulitis after open lacrimal surgery. Intraoperative administration of antibiotics has the advantages of compliance and economics. PMID- 10690833 TI - The surgical management of hypotony. AB - OBJECTIVE: To study the effect of dissecting epiciliary proliferative tissue in eyes that are hypotonous. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Eight patients (nine eyes). MAIN OUTCOME MEASURES: Normalization of intraocular pressure and preservation of vision. METHODS: A chart review was conducted to locate all surgical procedures performed primarily for hypotony. Nine procedures on eight eyes of eight patients seen in the vitreoretinal service of the Wilmer Ophthalmological Institute were identified and included in this study. Six of these eyes had undergone prior surgery for retinal detachment in association with proliferative vitreoretinopathy, one had undergone surgery for retinal detachment in the setting of a ruptured globe, and two had undergone cataract surgery and coined the diagnoses of uveitis (juvenile rheumatoid arthritis and sarcoid). Dissection and removal of the epiciliary proliferative tissue and lens capsule was undertaken by two surgeons (EdJ and MSH) using either a pars plana or limbal approach. The main outcome measures were intraocular pressure (IOP) and visual acuity after an average follow-up of 26 months (range, 9-48 months). RESULTS: In the immediate postoperative period, all patients showed an increase in IOP (average increase of 6.3 mm Hg; range, 3-14 mmHg). This average increase in IOP decreased to 5.2 mmHg at 6 months and 4.2 mm Hg at 12 or more months of follow-up. The rate of the IOP decrease appeared to lessen with time, suggesting long-term stabilization. Visual acuity remained stable in all patients, although the final level of vision was low. CONCLUSIONS: Surgical intervention for hypotony with greater than 1 year follow-up continues to show elevated IOP in some patients, despite the fact that the large initial increase in IOP is not sustained. No eye had phthisis develop or became painful. However, no eye had marked improvement in vision. Further study is needed to uncover the main contributing factors that result in hypotony before IOP can be maintained for prolonged periods in more hypotonous eyes and before these eyes can regain more vision. PMID- 10690834 TI - A comparison of dense versus less dense diode laser photocoagulation patterns for threshold retinopathy of prematurity. AB - OBJECTIVE: To determine if the density of diode laser photocoagulation for the treatment of zone 1 or zone 2 threshold retinopathy of prematurity (ROP) affects the rate of progression of the disease. DESIGN: Retrospective, nonrandomized, comparative trial (n = 12) and prospective, randomized, clinical trial (n = 46). PARTICIPANTS: Two surgeons treated a total of 107 eyes from 58 patients with zone 1 or zone 2 threshold ROP within 72 hours of diagnosis. The two consecutive groups of patients were treated with two different diode laser photocoagulation patterns between May 1995 and October 1997 and were observed for at least 3 months. INTERVENTION: All patients underwent diode laser photocoagulation of the peripheral avascular retina extending from the ridge of extraretinal proliferation to the ora serrata. One cohort received a near confluent laser pattern, whereas the second cohort received a pattern of laser spots placed 1 to 1.5 burn widths apart. MAIN OUTCOME MEASURES: Anatomic outcome, rate of progression to stage 4 or 5 retinopathy of prematurity, postoperative complications, and timing and frequency of retreatment. RESULTS: For analysis, the retrospective and randomized outcome data were grouped. The rate of progression in the near confluent laser treatment group was 3.6% overall, 0% of zone 1 eyes, and 3.8% of zone 2 eyes. The rate of progression in the less dense treatment group was 29% overall, 44% of zone 1 eyes, and 21 % of zone 2 eyes. Mean time to retreatment was 16 days in cohort 1 and 24 days in cohort 2. CONCLUSIONS: A dense pattern of diode laser treatment for threshold ROP and prompt retreatment for residual plus disease significantly reduce the rate of progression in eyes with zone 2 disease (P = 0.02) and may be beneficial in eyes with zone 1 disease. PMID- 10690835 TI - Laser photocoagulation spot-size errors stemming from the refractive state of the surgeon's eye. AB - PURPOSE: Meaningful errors in photocoagulation spot size may result from several factors. In this article we discuss one major factor, namely, fluctuations in the surgeon's accommodative state, coupled with an inaccurate setting of the slit lamp oculars. DESIGN: Experimental study. METHODS: We analyzed and tested the optics of slit-lamp mounted lasers. Varying the ocular setting is correlated with measurements of the actual spot size obtained with each system. MAIN OUTCOME MEASURE: The spot size obtained. RESULTS: Three distinct, but related, phenomena that may lead to spot size errors are defined: (1) focusing the laser spot as opposed to focusing the retinal image; (2) instrument misalignment; (3) inadvertent accommodation. CONCLUSION: The ocular setting must be meticulously calibrated to produce a true spot-sized burn. At the 50 microm setting, each diopter of induced accommodation, or erroneous ocular setting, almost doubles the actual spot size obtained. With large (500 microm) spot size settings, the defocused delivery system is more prone to spot-size errors in contrast with parfocal lasers. PMID- 10690836 TI - Peripapillary chorioretinal atrophy: Bruch's membrane changes and photoreceptor loss. AB - PURPOSE: To determine relationships among Bruch's membrane ultrastructure, photoreceptor degeneration, and choriocapillaris atrophy with respect to zones of retinal pigment epithelium (RPE) degeneration and atrophy adjacent to the optic nerve head, as a function of age. DESIGN: Human tissue study using clinicopathologic correlation. TISSUES: Eyes from patients 36 to 93 years of age lacking clinical evidence of glaucoma, optic nerve abnormalities, severe myopia, age-related macular degeneration, or other macular or peripapillary chorioretinal pathologic condition. METHODS: Sections through the retina-choroid complex at the temporal aspect of the optic nerve head were used for light microscopic histopathologic analysis (n = 17), electron microscopy (n = 9), carbonic anhydrase histochemical analysis (n = 7), and lipid histochemical analysis (n = 22). Retinal whole mounts were used for photoreceptor counts (n = 5). MAIN OUTCOME MEASURES: We determined the width of RPE degeneration and atrophy, the number of eyes with abnormalities of inner Bruch's membrane, and the number of rod and cone photoreceptors within 1 mm of the disc margin. We determined whether Bruch's membrane changes, photoreceptor degeneration, and choriocapillaris atrophy were associated with RPE degeneration and atrophy. RESULTS: All eyes had peripapillary RPE atrophy, degeneration, or both. The zone of RPE atrophy widened significantly after age 75. Thickening of inner Bruch's membrane and abnormalities of the RPE basal lamina were associated with degenerating and atrophic RPE in all eyes. The RPE basal lamina was narrow, reduplicated, or thickened as a basal laminar deposit. All eyes exhibited degeneration and loss of rods but not cones at the peripapillary termination of Bruch's membrane. Diminution of choriocapillaris coverage of Bruch's membrane was associated with RPE degeneration. Complete loss of the choriocapillaris was associated with RPE atrophy. CONCLUSIONS: Our results suggest that peripapillary chorioretinal atrophy is an age-related degeneration of the RPE-Bruch's membrane complex that resembles that found in the macula and periphery of normal eyes. PMID- 10690837 TI - Circumscribed choroidal hemangioma: characteristic features with indocyanine green videoangiography. AB - OBJECTIVE: To determine the characteristic features of indocyanine green videoangiography (ICG-V) of circumscribed choroidal hemangioma. DESIGN: Prospective, observational case series. PARTICIPANTS: Twenty-five eyes of 25 consecutive patients with circumscribed choroidal hemangioma. INTERVENTION: Indocyanine green videoangiography and intravenous fluorescein angiography (IVFA) were prospectively performed and reviewed. The specific features on ICG-V were compared with features of IVFA. MAIN OUTCOME MEASURES: The behavior of circumscribed choroidal hemangioma cases was observed with ICG-V and IVFA. RESULTS: On ICG-V, earliest hyperfluorescence of circumscribed choroidal hemangioma was achieved at a mean of 27.6 seconds (range, 13-62 seconds), whereas maximum hyperfluorescence occurred at 222 seconds (range, 33-707 seconds). In the late frames, all eyes demonstrated a relative decrease in fluorescence, including 18 eyes (72%) that demonstrated "washout" of the dye. Other findings on ICG-V included intrinsic vessels in 19 eyes (76%), a late hyperfluorescent rim in 19 eyes (76%), and late frame hot spots in 14 eyes (56%). On IVFA, the earliest hyperfluorescence was achieved at a mean of 24 seconds (range, 10-66 seconds), whereas maximum hyperfluorescence occurred at a mean of 76.3 seconds (range, 21 720 seconds). Increasing hyperfluorescence in the late frames was found in all cases. Other findings included intrinsic vessels in 12 eyes (48%) and hot spots in the late frames in 9 eyes (36%). CONCLUSIONS: Circumscribed choroidal hemangioma have specific characteristics on ICG-V that are not visualized with IVFA. We believe that ICG-V may become an important noninvasive tool for the diagnosis of choroidal hemangioma. PMID- 10690838 TI - Transpupillary thermotherapy for circumscribed choroidal hemangiomas. AB - PURPOSE: Various treatments have been described for circumscribed choroidal hemangiomas. We present our results with transpupillary thermotherapy (TTT) with infrared laser that indicate that the therapy is a new, effective treatment for this pathologic condition. DESIGN: Noncomparative, prospective, interventional case series. PARTICIPANTS: The treatment and follow-up of eight eyes of eight patients with circumscribed choroidal hemangiomas are presented. Each case was treated when the visual acuity (VA) decreased because of serous retinal detachment that affected the macula. METHODS: Infrared diode laser was used at 810 nm and power between 800 and 1200 mW with a beam diameter of 2 or 3 mm depending on the diameter of the hemangioma, with 3 to 6 minutes of exposure time. MAIN OUTCOME MEASURES: The final VA, the presence of serous subretinal fluid, funduscopic appearance of the tumor, and the degree of hyperfluorescence observed during fluorescein angiography were the main parameters. RESULTS: In all eight cases, total reabsorption of the serous retinal detachment was achieved after one or repeated applications of TTT. Mild choroidal atrophy and minimal hyperpigmentation of the retinal pigment epithelium were observed in the treated eyes. CONCLUSIONS: TTT can be considered an acceptable therapeutic option for choroidal hemangiomas and serous retinal detachment, and we believe that the role of this therapy will expand in the management of these patients. PMID- 10690839 TI - Stereotactic radiation therapy for malignant choroidal tumors: preliminary, short term results. AB - PURPOSE: To evaluate the role of stereotactic radiation therapy (SRT) in the treatment of malignant choroidal tumors. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Ten patients with unifocal choroidal metastasis (three lung carcinoma, three breast carcinoma, three colon carcinoma, one cutaneous melanoma) and five patients with primary choroidal melanoma underwent single-dose or fractionated SRT. METHODS: Before treatment, computed tomography (CT) scans of the orbit were obtained with the patient wearing an individualized immobilization mask. An integrated macro-CCD-camera system viewed the eye for detection of movements. Three-dimensional computer-based treatment planning was carried out. Dose distribution was calculated and displayed in isodose lines on the CT data set. For SRT, a dedicated stereotactic linear accelerator (6 MV) was used. Total doses for choroidal metastases were 12 to 20 Gy in a single dose or 30 Gy over 10 days (3 Gy each session), and total doses for choroidal melanoma were 50 Gy over 5 or 10 days (10 or 5 Gy each session). MAIN OUTCOME MEASURES: Best corrected visual acuity (ETDRS-chart), biomicroscopy, ultrasound examination, fluorescein angiography, and magnetic resonance imaging (MRI) were performed before treatment and at regular intervals after completion of SRT. RESULTS: During a follow-up period from 1 to 34 months (median, 6.5 months), local tumor control was achieved in all eyes. A decrease in tumor size on ultrasonography or MRI was noted in eight patients. No persistent side effects were observed during follow-up. CONCLUSIONS: Stereotactic radiation therapy allows steep dose gradients outside the target volume by minimizing the field of exposure. Thus only low radiation doses affect surrounding radiosensitive ocular structures. Our initial findings suggest that this technique may be effective in controlling tumor growth. Further studies are needed to compare treatment efficacy and safety with conventional treatment methods. PMID- 10690840 TI - Determination of surgeon's absorbed dose in iodine 125 and ruthenium 106 ophthalmic plaque surgery. AB - OBJECTIVE: Validation of dosimetry and exposition time to an ophthalmic surgeon during radioactive plaque operations. DESIGN: Experimental study in which videotaped operations and dosimetric measurements were used to model dosimetry. METHODS: We used thermoluminescence detectors for high sensitivity readings in radiation fields. Typical intersurgical mobility was videotaped and distances to the plaque was evaluated. MAIN OUTCOME MEASURE: Estimated radiation received by surgeons. RESULTS: All simulated plaque operations have a maximum dose rate of 6 mGy per minute (value in the inner eyeball). Mean dose rate is 2 mGy per minute (average of approximately all measurements). The surgeon's fingers receive a dose from 2 to 6 mSv from plaque operation. CONCLUSIONS: Results suggest that radioactive plaque operations are safe for the surgeon but that the time for plaque handling should be minimized. A surgeon should not exceed 100 to 200 operations per year. PMID- 10690841 TI - Endothelial cell loss in irradiated optic nerves. AB - OBJECTIVE: Radiation optic neuropathy usually occurs months to years after exposure of the anterior visual pathways to ionizing radiation. It is characterized by high signal on gadolinium-enhanced T1-weighted magnetic resonance imaging. Radiation-induced endothelial cell damage resulting in blood nerve barrier breakdown is hypothesized to produce this pattern, but histologic evidence of this in the optic nerve is lacking. We attempted to evaluate the effect of radiation on endothelial cells in the optic nerve. DESIGN: Case controlled histologic study. METHODS: We studied the optic nerves of 16 enucleated eyes from patients with uveal melanoma treated with proton beam irradiation, 6 from normal eyes and 5 from eyes with unirradiated uveal melanomas. Binding of Ulex europaeus agglutinin I (UEA-I) lectin was used to identify endothelial cells in single paraffin sections. Transverse and longitudinal sections of vessels were counted in masked fashion. RESULTS: There were 49.4+/-6.9 transversely sectioned endothelial cells per millimeter of nerve in 6 optic nerves exposed to 0 to 1000 cGyE ("low-dose") compared with 17.3+/-5.3 in 10 nerves exposed to 5500 to 7000 cGyE ("high-dose") (P = 0.002). Longitudinally sectioned vessels stained with UEA-I were separately identified, with 11.5+/-2.1 in the low-dose group and 5.6+/-1.6 in the high-dose group (P = 0.044). The thickness and staining of the endothelial cell layer appeared greater in the high-dose group. Endothelial cell counts did not correlate with age, gender, acuity, or interval after irradiation. CONCLUSIONS: Increased radiation dosage to the optic nerve correlates with smaller numbers of endothelial cells. PMID- 10690842 TI - Infrared scanning laser tomography of macular cysts. AB - OBJECTIVE: To perform three-dimensional, noninvasive, quantitative analysis of cystoid macular edema and macular cysts using infrared scanning laser tomography and to correlate findings with visual acuity (VA) as a basis for interventional studies. DESIGN: Cross-sectional, nonrandomized study. PARTICIPANTS: Seventeen patients (29-86 years of age) with macular cysts associated with a broad spectrum of diseases. INTERVENTION: Confocal infrared imaging with scanning laser tomography with the TopSS (790 nm) (Laser Diagnostic Technologies, San Diego, CA) with digitized images was used to perform three-dimensional, quantitative analysis of cysts in the central 5 degrees of the macula. MAIN OUTCOME MEASURES: Measurements of macular cyst number, area, volume, depth, slope, height of the surrounding macular elevation, and correlation with VA. RESULTS: Scanning laser tomography detected macular cysts in all patients. The number per patient ranged from 1 through 15. Cysts were accompanied by surrounding macular elevation in 16 patients (mean macular height, 216 microm). The area covered by cysts in the central 5 degrees was 0.087 to 0.969 mm2, and volume was 0.007 to 0.549 mm3. Visual acuity was significantly poorer in patients with greater cyst area (P = 0.0007), cyst volume (P = 0.0009), macular thickening (P = 0.0002), and cyst depth (P = 0.0013). Cyst number, average slope, and maximum slope, however, did not correlate significantly with VA. Grouping of macular cysts according to macular height and average cyst depth revealed that cysts in a more thickened retina were significantly deeper, had steeper slopes, and corresponded to worse VA. Macular height and average cyst depth were highly associated with each other, suggesting that in eyes with surrounding macular edema, cysts were deeper and may reflect more widespread tissue destruction. Individual confocal tomographic images provided additional information. Neither ophthalmoscopy nor fluorescein angiography delineated features such as retinal folds that suggested vitreous traction or changes in deeper layers that suggested occult choroidal new vessels. CONCLUSIONS: Infrared scanning laser tomography is a rapid and noninvasive imaging method that provides quantitative analysis of macular cysts in addition to qualitative information not seen clinically. Because poor VA is related to severe involvement of the central retina, scanning laser tomography could provide an objective outcome measure for interventional studies. PMID- 10690843 TI - A longitudinal study of visual function in carriers of X-linked recessive retinitis pigmentosa. AB - OBJECTIVE: This study was carried out to evaluate the progression of visual function impairment in carriers of X-linked recessive retinitis pigmentosa. We also assessed the relationship between the retinal findings at presentation and the extent of deterioration. DESIGN: Observational, retrospective, case series. PARTICIPANTS: Twenty-seven carriers of X-linked recessive retinitis pigmentosa. METHODS: Each carrier was clinically categorized into one of four grades (grades 0 through 3) depending on the presence or absence of a tapetal-like retinal reflex and the extent of peripheral pigmentary degeneration. A complete ophthalmologic examination was performed and data for visual acuity, visual field area, and electroretinographic measurements were collected on the most recent visit in both eyes. These were then compared with similar data obtained on their initial visits. MAIN OUTCOME MEASURES: A comparison of visual function was carried out between the initial visit and the most recent visit on each carrier. The visual acuity was measured with Snellen's acuity charts. The visual fields to targets V-4-e and II-4-e were planimeterized and used for the analysis. The electroretinographic (ERG) measures used were light-adapted single-flash b-wave amplitudes and 30-Hz red flicker for cone function, dark-adapted maximal b-wave amplitudes, and response to a low intensity blue-flash for rod function. RESULTS: None of the 11 carriers with a tapetal-like reflex only (grade 1) showed any significant change in visual acuity or fields as compared with 3 of 7 (43%) carriers with diffuse peripheral pigmentary findings (grade 3) who showed significant deterioration in visual acuity in at least one eye, and 6 of 7 (86%) who showed a significant decrease in visual field area with at least one target size in at least one eye. By comparison, only 1 of 10 carriers with a grade 1 fundus finding demonstrated a significant decrease in maximal dark-adapted ERG function as compared with 5 of 6 (83%) carriers with grade 3 in response to a single-flash stimulus and with 4 of 5 (80%) carriers in response to a single flash blue stimulus. For the single-flash photopic response, none of the 10 carriers with grade 1 showed any significant deterioration, whereas 2 of 4 (50%) with grade 3 did show such deterioration. The ERG responses for carriers with grade 2 were in between the extent of decrease in ERG amplitudes of those in carriers with grades 1 and 3. CONCLUSIONS: In our cohort of X-linked retinitis pigmentosa carriers, those with only a tapetal-like retinal reflex at presentation had a better prognosis to retain visual function than those with peripheral retinal pigmentation. These data are useful in counseling such carriers as to their visual prognosis. PMID- 10690844 TI - HSV-1--induced acute retinal necrosis syndrome presenting with severe inflammatory orbitopathy, proptosis, and optic nerve involvement. AB - OBJECTIVE: To present a unique case in which orbital inflammation, proptosis, and optic neuritis were the initial symptoms of acute retinal necrosis (ARN). The clinical presentation of ARN, as well as the currently recommended diagnostic procedures and guidelines for medical treatment of ARN, are summarized. DESIGN: Interventional case report. TESTING: Polymerase chain reaction (PCR) techniques were made on the vitreous for cytomegalovirus, Epstein-Barr virus, herpes simplex virus (HSV), varicella zoster virus, and toxoplasmosis. A full laboratory evaluation was made together with HLA-typing and serologic tests measuring convalescent titers for HSV and other micro-organisms. Magnetic resonance imaging scan, computed tomography (CT) scan, and fluorescein angiographic examination were performed. The patient was treated with acyclovir and oral prednisone. MAIN OUTCOME MEASURES: The patient was evaluated for initial and final visual acuity and for degree of proptosis, periocular edema, and vitreitis. RESULTS: The first symptoms and signs of ARN were eye pain, headache, proptosis, and a swollen optic nerve on CT scan. Other than increased C-reactive protein, all blood samples were normal. PCR was positive for HSV-type I in two separate vitreous biopsies. The patient had the strongly ARN-related specificity HLA-DQ7. CONCLUSIONS: This is the first report of HSV-induced ARN presenting with inflammatory orbitopathy and optic neuritis. Polymerase chain reaction for HSV-1 was positive more than 4 weeks after debut of symptoms, which is a new finding. The combination of severe vitreitis and retinal whitening, with or without proptosis, should alert the clinician to the possibility of herpes infection and treatment with intravenous acyclovir started promptly. PMID- 10690845 TI - Impact of large angle horizontal strabismus on ability to obtain employment. AB - OBJECTIVE: To determine if large angle esotropia and exotropia could impact a person's ability to obtain employment. DESIGN: Laboratory experiment. PARTICIPANTS: Seventy-nine respondents unaware of the purpose of the study. METHODS: Photographs of two men and two women were digitally altered to create photographs of the same individual in an orthotropic, esotropic, and exotropic state. The photographs were then randomly affixed to similarly qualified job resumes. The 79 study respondents, unaware of the purpose of the study, were asked to (1) rate each individual applicant on selected job qualification variables, and (2) rank the applicants against each other in order of hiring preference. MAIN OUTCOME MEASURES: Individual applicant rating and hiring preference scores. RESULTS: Women with normal ocular alignment received greater hiring preference scores than did strabismic women (P = 0.007). No difference in hiring preference scores was noted between strabismic and non-strabismic male applicants (P = 0.47). CONCLUSIONS: Large angle horizontal strabismus appeared to be vocationally significant for female applicants, reducing a strabismic female applicant's ability to obtain employment. The presence of strabismus did not appear to influence hiring decisions of male applicants. PMID- 10690846 TI - Clinical review 111: familial sex reversal: a review. PMID- 10690847 TI - Therapeutic controversy: prevention and treatment of diabetes in children. PMID- 10690848 TI - Reversal of hypoglycemia unawareness in a long-term type 1 diabetic patient by improvement of beta-adrenergic sensitivity after prevention of hypoglycemia. AB - The purpose of this study was to assess the effect of strict avoidance of hypoglycemia on beta-adrenergic sensitivity in a type 1 diabetic patient with hypoglycemia unawareness and a diabetes duration of 55 yr. beta-Adrenergic sensitivity was determined by an isoproterenol test and was expressed as the lowest dose of isoproterenol that increases the heart rate by 25 beats/min (IC25). Plasma epinephrine and symptom responses to hypoglycemia were determined during a 3-h hypoglycemic (3 mmol/L) clamp. Initially, the patient had a near normal counterregulatory plasma epinephrine response to hypoglycemia but reduced beta-adrenergic sensitivity (IC25, 2 microg) compared to 10 hypoglycemia aware, type 1 diabetic patients (0.65 +/- 0.14 microg) and 10 normal control subjects (1.13 +/- 0.21 microg). After 1 yr of strict avoidance of blood glucose levels below 4 mmol/L, the IC25 decreased to 0.25 microg, reflecting improved beta adrenergic sensitivity. In conclusion, the reduced beta-adrenergic sensitivity in this patient was probably the reason for hypoglycemia unawareness and was reversed by strict avoidance of hypoglycemia. PMID- 10690849 TI - Criteria for cure of acromegaly: a consensus statement. AB - In February 1999, a workshop was held in Cortina, Italy to develop a consensus defining the criteria for cure of acromegaly. The workshop was sponsored by the University of Brescia and hosted by the Italian Society of Endocrinology. Invited international participants included endocrinologists, neurosurgeons, and radiotherapists skilled in the management of acromegaly. This statement summarizes the consensus achieved in these discussions. PMID- 10690850 TI - Tumor necrosis factor-alpha inhibits leptin production in subcutaneous and omental adipocytes from morbidly obese humans. AB - This study was undertaken to examine the regulation of leptin production from human adipocytes by tumor necrosis factor-alpha (TNFalpha). Adipocytes were isolated from adipose tissue obtained during bariatric surgical procedures (17 women and 3 men; body mass index, 52.5 +/- 2.4 kg/m2; age, 40 +/- 3 yr) and cultured in suspension. Leptin release from sc adipocytes was inhibited 17.7 +/- 5.2% (P < 0.01), 21.6 +/- 4.3% (P < 0.005), and 37.1 +/- 7.2% (P < 0.05) by 1, 10, and 100 ng/mL TNFalpha, respectively, after 48 h in culture. At 100 ng/mL, significant inhibition of leptin release (25.8 +/- 9.7%; P < 0.05) was detected by 24 h. TNFalpha (10 ng/mL) had no effect on dexamethasone (0.1 micromol/L) stimulated leptin production in sc adipocytes. In omental adipocytes TNFalpha inhibited leptin release 21.0 +/- 9.6% and 40.8 +/- 6.3% at 10 and 100 ng/mL by 48 h (P < 0.05). Significant inhibition ofleptin release from omental adipocytes was observed at 24 h with 100 ng/mL TNFalpha (P < 0.05). Anti-TNFalpha antibody completely blocked TNFalpha inhibition of leptin release. The ob messenger ribonucleic acid was significantly reduced (23.6 +/- 5.9%) after 48 h of TNFalpha (100 ng/mL) treatment (P < 0.025). TNFalpha had no effect on glucose uptake or lactate production in sc and omental adipocytes. The data suggest that the direct paracrine effect of adipose-derived TNFalpha is inhibition of leptin production. PMID- 10690851 TI - A population-based study of chronic autoimmune hypothyroidism in Danish twins. AB - Hashimoto's thyroiditis (HT), atrophic thyroiditis (AT), and Graves' disease are autoimmune thyroid diseases in which genetic factors are suspected to play an important role in disease susceptibility. In a recent population-based twin study we rendered it probable that a substantial part of the susceptibility to Graves' disease is attributable to genetic factors. At present there are no population based twin studies supporting such a genetic influence in the etiology of HT/AT. To elucidate whether there is a genetic influence in the etiology of HT/AT, we studied the distribution of HT/AT in a population-based sample of 2945 Danish female-female twin pairs (5890 individuals) born between 1953 and 1972. Information on hypothyroidism was obtained from a nationwide questionnaire survey in 1994. Information from hospitals, out-patient clinics, general practitioners, and specialists was sought to verify the diagnosis. The overall prevalence of autoimmune hypothyroidism was 0.41% (24 of 5890). The prevalence did not differ between monozygotic and dizygotic twins (0.42% and 0.40%, respectively). The crude proband-wise concordance rates were significantly higher for monozygotic compared to dizygotic twin pairs: 0.55 (95% confidence interval, 0.23-0.83) vs. 0.0 (95% confidence interval, 0.0-0.25; P = 0.01). All of the healthy cotwins (n = 15) of twins with clinically overt autoimmune hypothyroidism were biochemically euthyroid. Overall, regardless of zygosity 53% (8 of 15) of the healthy cotwins were positive for antithyroid antibodies. The prevalence of autoantibodies among the monozygotic cotwins was 80% (4 of 5) and 40% (4 of 10) among dizygotic cotwins (P = 0.36). In conclusion, the higher concordance rate in monozygotic compared to dizygotic pairs indicates that genetic factors play a role in the etiology of HT/AT among Caucasian women living in areas with borderline iodine deficiency. However, the fact that the concordance rate among MZ twins was below 1 suggests that environmental factors also are of etiological importance. PMID- 10690852 TI - Detection of minimal levels of serum anti-Mullerian hormone during follow-up of patients with ovarian granulosa cell tumor by means of a highly sensitive enzyme linked immunosorbent assay. AB - Granulosa cell tumors (GCT) are ovarian neoplasms that tend to recur and spread in the pelvis and the abdomen several years after the initial treatment. Anti Mulerian hormone (AMH) is a reliable serum marker of these tumors. To enhance the availability and the sensitivity of serum AMH determination, we developed an ultrasensitive enzyme-linked immunosorbent assay. In this work we compare the results of serum AMH levels, obtained using the ultrasensitive and the traditional assays, in 31 patients with ovarian GCT followed up for up to 7 yr. The ultrasensitive enzyme-linked immunosorbent assay has a significantly higher sensitivity than the traditional one. This resulted in the detection of low serum AMH levels, which were undetectable with the traditional assay, in several cases including one patient in whom a recurrence of a GCT had developed and two patients in whom the treatment had not been completely successful. These cases highlight the importance of the availability of a highly sensitive assay allowing evaluation with high precision of the results of treatment and to detect the recurrences of GCT at an early, preclinical stage. PMID- 10690853 TI - Decrease of free thyroxine levels after controlled ovarian hyperstimulation. AB - Controlled ovarian hyperstimulation could lead to opposing effects on thyroid function. Therefore, in a prospective study of 65 women undergoing controlled ovarian hyperstimulation, thyroid hormones, T4-binding globulin, TPO antibodies, gonadotropins, estradiol, and PRL were measured before and after controlled ovarian hyperstimulation. After ovarian stimulation (mean +/- SE of mean): free T4 decreased, 14.4 +/- 0.2 vs. 12.9 +/- 0.2 pmol/L (P < 0.0001); thyroid stimulating hormone increased, 2.3 +/- 0.3 vs. 3.0 +/- 0.4 mU/L (P < 0.0001); T4 binding globulin increased, 25.2 +/- 0.7 vs. 33.9 +/- 0.9 mg/L (P < 0.0001); total T4 increased, 98.1 +/- 2.3 vs. 114.6 +/- 2.5 nmol/L (P < 0.0001); total T3 increased, 2.0 +/- 0.04 vs. 2.3 +/- 0.07 nmol/L (P < 0.0001); TPO antibodies decreased, 370 +/- 233 U/mL vs. 355 +/- 224 U/mL (P < 0.0001); LH decreased, 8.1 +/- 1.1 vs. 0.4 +/-0.1 U/L (P < 0.0001); FSH did not change, 6.5 +/- 0.6 vs. 7.9 +/- 0.9 U/L (P = 0.08); human CG increased, <2 +/- 0.0 vs. 195 +/- 16 U/L (P < 0.0001); estradiol increased, 359.3 +/- 25.9 pmol/L vs. 3491.8 +/-298.3 pmol/L (P < 0.0001); and PRL increased, 0.23 +/- 0.02 vs. 0.95 +/- 0.06 U/L (P < 0.0001). Because low maternal free T4 and elevated maternal thyroid-stimulating hormone levels during early gestation have been reported to be associated with impaired psychomotor development in the offspring, our findings indicate the need for additional studies in the children of women who where exposed to high levels of estrogens around the time of conception. PMID- 10690854 TI - Use of long-term intravenous phosphate infusion in the palliative treatment of tumor-induced osteomalacia. AB - Tumor-induced osteomalacia is characterized by paraneoplastic defects in vitamin D metabolism, proximal renal tubular functions, and phosphate transport. The resulting hypophosphatemia can cause generalized pain and muscle weakness, which significantly affect the quality of life of the patients. Palliative treatment with calcium, vitamin D, and phosphate replacement is indicated for patients in whom the causative tumor cannot be completely resected. In this report we describe a case of tumor-induced osteomalacia in whom adequate oral doses of phosphate could not be used because of gastrointestinal side-effects. Long term (3-6 months) iv phosphate infusion delivered by ambulatory infusion pumps in combination with oral calcium and vitamin D was used successfully to decrease pain and increase muscle strength. Careful monitoring of serum calcium, phosphate, and creatinine levels and reliable microinfusion technology have allowed the long term use of iv phosphate infusion without serious morbidity. This patient received repeated (three times) phosphate infusions over 8 yr, resulting in laboratory and symptomatic improvement after each course. However, this patient did suffer two episodes of central venous catheter-related infection. Because of potentially serious complications, such as severe hypocalcemia, calcified right ventricular thrombi, and nephrocalcinosis, long term iv phosphate infusion should be reserved for patients who cannot tolerate adequate doses of oral phosphate and for whom the benefits outweigh the risks. PMID- 10690855 TI - Successful use of pulsatile gonadotropin-releasing hormone (GnRH) for ovulation induction and pregnancy in a patient with GnRH receptor mutations. AB - GnRH receptor mutations have recently been identified in a small number of familial cases of nonanosmic hypogonadotropic hypogonadism. In the present report we studied a kindred in which two sisters with primary amenorrhea were affected with GnRH deficiency due to a compound heterozygote mutation (Gln(106)Arg, Arg(262)Gln) and performed extensive phenotyping studies. Baseline patterns of gonadotropin secretion and gonadotropin responsiveness to exogenous pulsatile GnRH were examined in the proband. Low amplitude pulses of both LH and free alpha subunit (FAS) were detected during 24 h of every 10 min blood sampling. The proband then received exogenous pulsatile GnRH i.v. for ovulation induction, and daily blood samples for gonadotropins and sex steroids were monitored. At the conventional GnRH replacement dose for women with hypogonadotropic hypogonadism (75 ng/kg), no follicular development occurred. At a GnRH dose of 100 ng/kg, the level and pattern of gonadotropin secretion more closely mimicked the follicular phase of normal women; a single dominant follicle was recruited, and an endogenous LH surge was elicited. However, the luteal phase was inadequate, as assessed by progesterone levels. At a GnRH dose of 250 ng/kg, the gonadotropin and sex steroid dynamics reproduced those of normal ovulatory women in both the follicular and luteal phases, and the proband conceived. The FAS responses to both conventional and high dose GnRH were within the normal range. The following conclusions were made: 1) Increased doses of GnRH may be used effectively for ovulation induction in some patients with GnRH receptor mutations. 2) Higher doses of GnRH are required for normal luteal phase dynamics than for normal follicular phase function. 3) Hypersecretion of FAS in response to exogenous GnRH, which is a feature of congenital hypogonadotropic hypogonadism, was not seen in this patient with a GnRH receptor mutation. PMID- 10690856 TI - Anemia in children with cartilage-hair hypoplasia is related to body growth and to the insulin-like growth factor system. AB - Cartilage-hair hypoplasia (CHH) is a metaphyseal chondrodysplasia characterized by severe short-limbed short stature, hypoplastic hair, and defective immunity. The patients also have anemia. As GH may regulate both body growth and erythropoiesis, we used CHH as a clinical model to study their interrelationships. Retrospective analysis of hematological data of 114 patients showed that the severity of the anemia and macrocytosis in CHH varies with age. The anemia was most severe in early childhood. A prospective study of 21 patients with CHH showed that height correlates with hemoglobin (P = 0.006) and mean corpuscular volume of red blood cells (P < 0.0001). The individual hemoglobin levels correlated with the GH parameters [P = 0.035 for insulin-like growth factor I (IGF-I) and P = 0.002 for IGF-binding protein-3], and the mean corpuscular volume of red blood cell values correlated with fetal hemoglobin. Bone marrow cultures obtained from six patients with CHH showed reduced or totally absent erythroid colony formation, which was not influenced by GH or IGF I in vitro or by GH treatment in vivo. In patients with CHH, we observed an association between erythropoiesis and growth. We conclude that body growth and erythropoiesis share common regulators. One of these is the GH-IGF-I axis; other factors, as not yet identified, may also be important. PMID- 10690857 TI - Near final height in pubertal growth hormone (GH)-deficient patients treated with GH alone or in combination with luteinizing hormone-releasing hormone analog: results of a prospective, randomized trial. AB - To study the effects of delaying puberty in GH-deficient (GHD) children, we studied 21 GHD (9 boys, 14 girls), treatment-naive, pubertal patients in a prospective, randomized trial. Their chronological age was 14.3 +/- 1.6 yr, and their bone age was 11.3 +/- 1.1 yr (mean +/- SD) at the beginning of the study. Four patients who developed hypogonadotropic hypogonadism were subsequently excluded from the study. Patients were randomly assigned to receive GH + LH releasing hormone analog (LHRH-A) (n = 7), or GH alone (n = 10). GH and LHRH-A treatment started simultaneously in each patient. GH (Nutropin) was administered at a dose of 0.1 U/kg x day sc, until patients reached a bone age (BA) of 14 yr in girls and 16 yr in boys, and LHRH-A (Lupron depot) was administered at a dose of 300 microg/ kg every 28 days in during 3 yr. We defined GH deficiency as patients with a growth velocity less than 4 cm/yr, BA delay more than 1 yr in relationship to chronological age, GH response to two stimulation tests less than 7 microg/L, associated with low serum insulin-like growth factor I and insulin like growth factor binding protein 3 levels. Statistical analysis was performed by ANOVA or Kruskall Wallis when variances were not homogeneous. We observed a significant decrease in the rate of BA maturation in the group treated with GH+LHRH-A (1.5 +/- 0.2 yr) compared with the group treated with GH alone (4.2 +/ 0.5 yr) during the 3 years of LHRH-A therapy (P < 0.05). This delay in BA maturation produced a significant gain in final height in the group treated with GH+LHRH-A, which reached - 1.3 +/- 0.5 SD score compared with -2.7 +/- 0.3 SD score (P < 0.05) in the group treated with GH alone. These results indicate that delaying puberty with LHRH-A in GHD children during treatment with GH increases final height. PMID- 10690858 TI - Hypopituitary females have a high incidence of cardiovascular morbidity and an increased prevalence of cardiovascular risk factors. AB - We recently reported that female patients with hypopituitarism receiving controlled thyroid and steroid hormone substitution, but without GH replacement, had a more than 2-fold increase in cardiovascular mortality compared to the general population. In the present study we investigated the incidence of cardiovascular disease as well as the prevalence of cardiovascular risk factors in 33 females with hypopituitarism for 6-46 yr (median, 18) compared to those in 33 control subjects recruited from the general population in the same geographical area and matched for sex, age, smoking habits, educational level, and residence location. The patients were with a very high probability GH deficient, as 29 had subnormal serum insulin-like growth factor I levels, and the other 4 were GH deficient, as assessed by an insulin tolerance test. The incidence of cardiovascular disease was significantly higher among the hypopituitary patients (incidence ratio, 3.7; 95% confidence interval, 1.2-11.3), and the consumption of cardioactive drugs was also significantly higher (P = 0.002). Hypopituitary patients had a lower degree of physical exercise during their spare time (P = 0.02), a higher waist/hip ratio (P = 0.01), lower high density lipoprotein cholesterol (P = 0.002), and higher low density/high density lipoprotein ratio (P = 0.009). Furthermore, the patients had a significantly increased left atrium size (P = 0.05), but no difference was observed for other cardiac measures. In the patients, serum insulin-like growth factor I levels significantly correlated with left ventricular mass index (r = 0.48; P = 0.006), suggesting that GH has a strong impact on cardiac size. More episodes of bradycardia (P = 0.05), but no increased occurrence of extrasystolies, were encountered in the patients during 24-h continuous electrocardiogram monitoring. Carotid artery intima-media thickness and plaque numbers did not differ between patients and controls. In conclusion, hypopituitary females exhibit an increased incidence of cardiovascular disease, higher cardioactive drug consumption, and an increased prevalence of cardiovascular risk factors. The increased cardiovascular morbidity could not be ascribed to inadequate estrogen or thyroid hormone treatment, and unsubstituted GH deficiency is probably an important contributing factor. PMID- 10690859 TI - Decreased prorenin processing develops before autonomic dysfunction in type 1 diabetes. AB - It is well documented that diabetic patients with chronic complications have decreased renin secretion and elevations in the renin precursor prorenin. It is uncertain, however, whether the abnormal processing of prorenin is reflective of microvascular disease, hypertension, or autonomic neuropathy. Dechaux et al. (Transplant Proc. 18:1598-1599, 1986) observed abnormalities in prorenin processing in uncomplicated diabetes and suggested that it was the result of subclinical autonomic neuropathy. To test this hypothesis, we measured renin, prorenin, and autonomic function in early type 1 diabetes at a time when there is little or no microvascular disease or hypervolemia. Thirty-seven patients (10 males, 27 females) enrolled 2-22 months after diagnosis in a longitudinal study in which renin, prorenin, and autonomic function were measured annually for 3 years. Forty-one age-matched control subjects were also studied. PRA in the diabetic patients at the time of the second and third evaluations was 1.71 +/- 0.24 ng angiotensin I/mL x h and 1.67 +/- 0.24 ng angiotensin I/mL x h, respectively, significantly lower (P < 0.05) than that of the control subjects in whom PRA was 2.96 +/- 0.38 ng angiotensin I/mL x h. Prorenin was not different in the diabetic patients in comparison with controls. The renin to prorenin ratio in the diabetic patients at the time of the first, second, and third evaluations was 0.260 +/- 0.03, 0.235 +/- 0.05, and 0.227 0.05, respectively, significantly lower (P < 0.01) than in control subjects in whom the renin to prorenin ratio was 0.475 +/- 0.08. Despite this, at the time of the first and second evaluations, there was no evidence of autonomic dysfunction and no correlation between any test of autonomic function and the renin to prorenin ratio. At the time of the third evaluation, however, the intermediate frequency (0.04-0.15 Hz) power spectra while patients were supine (an index of sympathetic modulation of heart rate variability) showed a highly significant (P < .001) correlation with the renin to prorenin ratio. High frequency (0.15-0.40 Hz) spectra from supine patients at the third evaluation also correlated with the renin to prorenin ratio (P < 0.01). We conclude abnormal processing of prorenin develops in diabetic patients prior to microvascular disease, even before the first evidence of autonomic dysfunction. Although the latter may play a contributory role, additional as yet unidentified mechanisms seem to interrupt the processing of prorenin in early diabetes. PMID- 10690860 TI - The influence of thinness and smoking on bone loss and response to hormone replacement therapy in early postmenopausal women. AB - We studied the influence of thinness and smoking in 153 early postmenopausal women from a 3-yr period, comparing treatments of 1 or 2 mg estradiol with placebo. The baseline body mass index (BMI) was significantly associated with bone resorption (r = -0.26, P < 0.01 for urinary CrossLaps with 21% difference between extreme tertiles) with a consequent association between BMI and bone mineral density (BMD; r = 0.38, P < 0.001 for BMD of hip with 10% difference between extreme tertiles). A low BMI led to an increased rate of loss (r = 0.45, P < 0.01 for BMD of spine), whereas response to treatment with either 1 or 2 mg estradiol was independent of BMI. Smoking was associated with a 4% lower BMD at baseline compared with that in nonsmokers. This effect was additive with that of BMI. For smokers the increase in serum estradiol during hormone replacement therapy was only half of that in nonsmokers. For women treated with placebo or 2 mg estradiol, serum FSH levels were similar in smokers and nonsmokers, but during treatment with 1 mg estradiol, serum FSH was significantly less suppressed in smokers. This was mirrored in the BMD response, where smokers responded similarly to 2 mg estradiol and placebo as did nonsmokers, whereas smokers receiving 1 mg estradiol experienced only half the increase compared to nonsmokers. In conclusion, thinness and smoking are important risk factors for osteoporosis development, but are counteracted by hormone replacement therapy. PMID- 10690861 TI - Carrier analysis and prenatal diagnosis of congenital adrenal hyperplasia caused by 21-hydroxylase deficiency in Chinese. AB - Congenital adrenal hyperplasia (CAH) is a common autosomal recessive disorder mainly caused by defects in the steroid 21-hydroxylase (CYP21) gene. We screened 1,000 healthy people, using a previously developed differential PCR method combined with single-strand conformation polymorphism and amplification-created restriction site methods for the carrier detection of the CYP21 gene deficiency. Our results indicated that the rate of occurrence of the heterozygous CAH carrier was about 12 in 1,000, with a gene frequency of 0.0060 and an incidence frequency of 1 in 28,000 in the Chinese population. In addition, 9 cases of CAH families were performed with prenatal diagnosis. Among them, 3 cases were diagnosed as the severe form, 4 cases carried the heterozygous mutation, and 2 were normal. This is the first report of carrier frequency analysis and prenatal diagnosis of 21 hydroxylase deficiency in Chinese. PMID- 10690862 TI - Exogenous 20K growth hormone (GH) suppresses endogenous 22K GH secretion in normal men. AB - The physiological and pharmacological functions of the 20-kDa human GH (20K-hGH) isoform are unknown. We conducted a pharmacokinetic study of recombinant 20K-hGH in human subjects (Phase I clinical trial). Placebo or 20K-hGH was administered sc to normal men (20-31 yr of age, n = 6-8 per group) at 2100 h. Serum 20K- and 22K-hGH levels were monitored every 30 min for 24 h by specific enzyme-linked immunosorbent assays. Serum free fatty acid, insulin-like growth factor I, insulin, and glucose levels were measured for 24 h. In the placebo group, the secretion profiles of endogenous 20K- and 22K-hGH were pulsatile and similar to each other. The proportion of 20K- to 22K-hGH was fairly constant. In the 20K-hGH treated groups, serum 20K-hGH levels increased in a dose-dependent manner over the dose range of 0.01-0.1 mg/kg. Maximum serum 20K-hGH levels were reached at 3 4 h and decreased with half-lives of 2-3 h. Marked suppression of endogenous 22K hGH secretion was observed in a time-dependent manner. Serum free fatty acid and insulin-like growth factor I levels were significantly elevated (P < 0.01) at 4, 8, and 12 h and at 8, 12, and 24 h after 20K-hGH administration, respectively. Serum insulin and glucose levels did not change significantly within 24 h. These results suggested that: 1) regulation of 20K-hGH secretion is physiologically the same as that of 22K-hGH; 2) the pharmacokinetics after sc injection of 20K-hGH are comparable with those of 22K-hGH; 3) 20K-hGH regulates hGH secretion through "GH-induced negative feedback mechanisms"; and 4) administration of 20K-hGH is expected to exert GH actions (growth-promoting activity and lipolytic activity). Monitoring of serum 20K- and 22K-hGH levels may be useful in evaluating the effects of administered GH isoforms on their own release from the pituitary. PMID- 10690863 TI - Relationship between serum inhibin A and B and ovarian follicle development after a daily fixed dose administration of recombinant follicle-stimulating hormone. AB - The aim of this study was to investigate the relationship of serum inhibin A and inhibin B to ovarian follicular development in women undergoing pituitary down regulation and ovarian stimulation with a fixed daily dose of recombinant human FSH in an in vitro fertilization program. Thirty-eight patients were treated randomly with either 100 or 200 IU/day recombinant human FSH (Puregon) for a period of 9-14 days. Serum FSH, inhibin A, inhibin B, 17beta-estradiol, and follicular size and number were determined before FSH treatment and every second day from days 4-6 throughout FSH treatment. Serum FSH increased in a dose-related manner to reach a maximum by days 4-6 and remained unchanged over the duration of treatment. Serum inhibin A and 17beta-estradiol also increased with increasing FSH dose and continued to rise throughout the FSH treatment period. By contrast, serum inhibin B was increased by days 4-6 at both doses of FSH to reach a maximum by days 7-8, remaining unchanged thereafter. Serum inhibin B and, to a lesser extent, inhibin A correlated significantly with the number of oocytes retrieved even when assessed early (days 4-6) in the treatment period (inhibin B vs. number of oocytes: r = 0.89; P < 0.001; inhibin A vs. number of oocytes: r = 0.61; P < 0.05). Serum inhibin A, inhibin B, and 17beta-estradiol were weakly correlated with the number of follicles less than 11 mm when assessed on a daily basis; stronger correlations were observed with the greater than 11-mm follicles during the late stages of treatment. It is concluded that serum inhibin B levels determined during the early stages (e.g. days 4-6) of fixed dose FSH treatment provide an early indicator of the number of recruited follicles that are destined to form mature oocytes. In this context, serum inhibin B may be of predictive value in monitoring ovarian hyperstimulation treatment for in vitro fertilization. PMID- 10690864 TI - The effect of hormone replacement therapy on cardiovascular hemodynamics in women with Turner's syndrome. AB - Women with Turner's syndrome, the majority of whom are estrogen deficient, have an increased incidence of coronary artery disease. The aim of this study was to assess the effects of hormone replacement therapy (HRT) on central arterial hemodynamics, insulin sensitivity, and lipids in adults with Turner's syndrome. Twenty-one women with Turner's syndrome were studied prospectively, on and off 3 months of estradiol valerate in combination with levonorgestrel. The following measurements were made: body mass index, waist/hip ratio, serum lipids, fasting insulin and glucose, and mean arterial blood pressure. Aortic root pressure and waveforms were estimated noninvasively and the augmentation index (AI), a measure of aortic stiffness, was calculated. The AI was significantly lower during estrogen therapy (22% vs. 15%; P = 0.008), suggesting a reduction in central arterial stiffness. Fasting insulin and glucose concentrations were also significantly lower during HRT (P = 0.01 and P = 0.0004, respectively). There was no difference in body mass index, serum lipids, or brachial and aortic blood pressures on and off treatment. Total cholesterol was correlated with the AI (r = 0.4; P = 0.03). These results suggest that HRT in women with Turner's syndrome has a favorable effect on central arterial hemodynamics and insulin sensitivity. The lack of effect on serum lipids suggests that the effects of HRT on aortic compliance may be mediated by an improvement in endothelial function. PMID- 10690865 TI - Adult height in short normal girls treated with gonadotropin-releasing hormone analogs and growth hormone. AB - Combined treatment with GH and GnRH analogs (GnRHa) has been proposed to improve final adult height in true precocious puberty, GH deficiency, and short normal subjects with early or normal timing of puberty with still controversial results. We treated 12 girls with idiopathic short stature and normal or early puberty with GH and GnRHa and followed them to adult height; 12 girls comparable for auxological and laboratory characteristics treated with GH alone served to better evaluate the efficacy of addition of GnRHa. At the start of combined treatment, the chronological age of the girls (CA; mean +/- SD) was 10.2 +/- 0.9 yr, bone age (BA) was 10.6 +/- 1.9 yr, height SD score for BA was - 1.81 +/- 0.8, PAH was 146.3 +/- 5.0 cm. PAH was significantly lower than target height (TH 152.7 +/- 3.6 cm; P < 0.005). GH was given at a dose of 0.3 mg/kg x week, sc, 6 days weekly, and GnRHa (depot-triptorelin) was given at a dose of 100 microg/kg every 21 days, im. The 12 girls were treated with GH alone at the same dose; at the start of therapy their CA was 10.7 +/- 1.0, BA was 10.1 +/- 1.4 yr, height SD score for BA was - 1.65 +/- 0.8, PAH was 145.6 +/- 4.4 cm, and TH was 155.8 +/- 4.6 cm. Pubertal Tanner stage in both groups was B2P2 or B3P3. LHRH test and pelvic ultrasound showed the beginning of puberty. The GH response to standard provocative tests was 10 g/L or more. The mean period of treatment was 4.6 +/- 1.7 yr in the group treated with GH plus GnRHa and 4.9 +/- 1.4 yr in the group treated with GH alone; both groups discontinued treatment at comparable CA and BA. Adult height was considered to be attained when growth during the preceding year was less than 1 cm, with a BA of over 15 yr. Patients in the group treated with GH plus GnRHa showed an adult height significantly higher (P < 0.001) than the pretreatment PAH (156.3 +/- 5.9 vs. 146.3 +/- 5 cm); the gain in centimeters calculated between pretreatment PAH and adult height was 10 +/- 2.9 cm, and 7 of 12 girls had a gain over 10 cm. Target height was significantly exceeded. Height SD score for BA increased from - 1.81 +/-0.8 to -0.85 +/- 1.0. The GH alone group reached an adult height higher than the pretreatment PAH (151.7 +/- 2.7 vs. 145.6 +/- 4.4 cm); the gain in final height vs. pretreatment PAH was 6.1 +/- 4.4 cm, and 5 of 12 girls did not gain more than 4 cm. TH was even not reached. The height SD score did not significantly change. No adverse effects were observed in either group. All of the girls showed good compliance and were satisfied with the results. Our experience suggests that the combination of GH and GnRHa is significantly more effective in improving adult height than GH alone in girls with idiopathic short stature, early or normal onset of puberty, and low PAH well below the third percentile and TH. As the cost-benefit of such invasive treatment must be seriously considered, further studies are needed due to the small sample of our patients as well as in other studies reported to date. PMID- 10690866 TI - Opposite variations in maternal and neonatal thyroid function induced by iodine supplementation during pregnancy. AB - Whereas the consequences of extremes in iodine intake are well described, much less is known about the effect of more moderate variations in maternal iodine intake on fetal thyroid function. The present study performed in Denmark with mild to moderate iodine deficiency dealt with the effect of maternal iodine supplementation on thyroid function in the mother at term and in the fetus/neonate. Serum was collected consecutively from pregnant women at term (n = 144) and from cord blood (n = 139). Forty-nine women had a regular intake of vitamin and mineral tablets with iodine (150 microg/day) during pregnancy, and 95 took no artificial iodine supplementation. Iodine supplementation (+I) induced opposite variations in thyroid function in the mother and the fetus. In +I mothers, TSH was 7.6% lower than in mothers with no supplementation (P < 0.05). In cord blood, on the contrary, TSH was 27.3% higher in the +I group (P < 0.05). The variations were caused by opposite shifts in TSH frequency distribution in mothers and neonates. The association between iodine supplementation and high serum TSH in the neonates was further substantiated by an inverse correlation between thyroglobulin and TSH in cord blood (P < 0.001), whereas no specific pattern was observed in the mothers. High serum thyroglobulin was a marker of low iodine intake in both mothers and neonates. The results suggest that the fetal thyroid, at least in areas of mild iodine deficiency, is more sensitive to the inhibitory effect of iodine than hitherto anticipated. PMID- 10690867 TI - Abnormal heart rate variability in adults with growth hormone deficiency. AB - GH-deficient (GHD) patients have increased risk of cardiovascular death and may have cardiac structural abnormalities. In non-GHD patients these are associated with cardiac autonomic dysfunction, and it is possible that autonomic dysfunction is also present in GHD patients. Power spectral analysis (PSA) of heart rate variability (HRV) indirectly measures cardiac autonomic tone and generates peaks at 3 frequency bands, very low frequency (VLF), low frequency (LF) and high frequency (HF). The area under the LF curve is considered to reflect predominantly cardiac sympathetic activity, whereas HF indicates parasympathetic activity. PSA of HRV was performed in 14 normotensive GHD patients (5 men and 9 women; mean age, 35.2 yr) and 19 healthy controls (9 men and 10 women; mean age, 38.3 yr). GHD patients had 26% lower normalized LF power (P < 0.004), 39% higher normalized HF power (P < 0.001), 28% lower normalized VLF power (P < 0.046), and 51% lower LF/HF ratio (an index of sympathovagal balance; P < 0.001) compared to controls. These data indicate that heart rate variability is abnormal in patients with GHD. The decreased sympathetic tone could be a consequence of reduced central sympathetic tone or altered cardiac responsiveness to autonomic control and may contribute to the increased cardiovascular risk in GHD patients. PMID- 10690868 TI - Growth hormone replacement therapy is not associated with retinal changes. AB - GH and/or growth factors are thought to play a role in the pathogenesis of diabetic retinopathy. In addition, the occurence of retinal changes mimicking diabetic retinopathy in two GH-deficient (GHD) patients receiving GH replacement therapy (GHRT) has recently been reported. The present study was performed to evaluate whether this was a coincidence or whether GHRT might regularly induce retinal changes. Sixty-one GHD patients on GHRT with a mean age of 42.5 +/-17.3 yr were examined by one ophthalmologist (AR). The mean duration of GHRT was 8.4 +/- 3.7 yr in childhood onset and 3.5 +/- 2.1yr in adult onset patients. Plasma insulin-like growth factor I concentrations were 76.4 +/- 49.6 ng/mL before GHRT and 244.3 +/- 119.2 ng/mL while receiving GHRT with a dose of 1.7 +/- 0.7 IU/day. After pupil dilatation with tropicamide, fundus examinations of both eyes were performed using a Volk 90 diopter fundus lens with a slit lamp (Haag Streit, Bern, Switzerland). In none of the patients were vascular or retinal changes like macular edema, microaneurysms, hemorrhages, hard exsudates, cotton wool spots, preproliferative signs, or proliferations found. The optic discs were also normal in all patients. We conclude, therefore, that long-term GHRT can be administered safely in GHD patients without an increased risk of retinal changes. PMID- 10690869 TI - A survey on adrenal incidentaloma in Italy. Study Group on Adrenal Tumors of the Italian Society of Endocrinology. AB - The aim of this study was to perform a national survey on occasionally discovered adrenal masses [adrenal incidentalomas (AI)] under the auspices of the Italian Society of Endocrinology. This multicentric and retrospective evaluation of patients with AI includes 1096 cases collected in 26 centers between 1980 and 1995. Relevant information was obtained by means of a specifically tailored questionnaire. Of the 1096 forms received, 1004 were retained for final analysis. Patients were 420 males and 584 females, aged between 15-86 yr (median, 58 yr). Mass size (computed tomography measurement) ranged from 0.5-25 cm (median, 3.0 cm). Hormonal work-up demonstrated that 85% of the masses were nonhypersecretory, 9.2% were defined as subclinical Cushing's syndrome, 4.2% were pheochromocytomas, and 1.6% were aldosteronomas. Adrenalectomy was performed in 380 patients with removal of 198 cortical adenomas (52%), 47 cortical carcinomas (12%), 42 pheochromocytomas (11%), and other less frequent tumor types. Patients with carcinoma were significantly younger than patients with adenoma (median, 46; range, 17-84; vs. 57, 16-83 yr; P = 0.05). Adenomas were significantly smaller than carcinomas (3.5, 1-15 vs. 7.5, 2.6-25 cm; P < 0.001), and a cut-off at 4.0 cm had the highest sensitivity (93%) in differentiating between benign and malignant tumors. Hormonal work-up of patients with subclinical Cushing's syndrome showed low baseline ACTH in 79%, cortisol unsuppressibility after 1 mg dexamethasone in 73%, above normal urinary free cortisol in 75%, disturbed cortisol rhythm in 43%, and blunted ACTH response to CRH in 55%. Only 43% of patients with pheochromocytoma were hypertensive, and 86% showed elevated urinary catecholamines. All patients with aldosteronoma were hypertensive and had suppressed upright PRA. These results indicate that mass size is the most reliable variable in separating benign from malignant AI. Adrenalectomy should be recommended for AI greater than 4.0 cm because of the increased risk of malignancy, especially in young patients. Endocrine evaluation should be performed in all patients to identify silent states of hormone excess. PMID- 10690870 TI - Hysterectomy, oophorectomy, and endogenous sex hormone levels in older women: the Rancho Bernardo Study. AB - This study examines the cross-sectional association of hysterectomy and oophorectomy status, chronological age, and years since menopause with plasma levels of total and bioavailable testosterone and estradiol, androstenedione, estrone, and sex hormone-binding globulin (SHBG) in community-dwelling postmenopausal women who were not using estrogen replacement therapy. Six hundred and eighty-four women, aged 50-89 yr, were surveyed for hysterectomy and oophorectomy status and had plasma obtained between 1984-1987. Of these, 438 (67%) had not undergone hysterectomy or oophorectomy (intact), 123 (18%) reported hysterectomy with bilateral oophorectomy, and 123 (18%) reported hysterectomy with conservation of 1 or both ovaries. After adjustment for age and body mass index, both total and bioavailable testosterone levels were reduced by more than 40% (P < 0.001) in hysterectomized women with bilateral oophorectomy compared to those in intact women, with intermediate levels observed in hysterectomized women with ovarian conservation. Androstenedione levels were about 10% lower in hysterectomized women with or without ovarian conservation compared to those in intact women (P = 0.039). Total estradiol levels tended to be lower (P = 0.095) in bilaterally oophorectomized women. Levels of bioavailable estradiol, estrone, and SHBG did not differ by hysterectomy and oophorectomy status. Among intact women, total, but not bioavailable, testosterone levels increased with age (P = 0.015), reaching premenopausal levels for the 70-79 decade with relatively stable levels thereafter. Among oophorectomized women, total and bioavailable testosterone levels did not vary with age and were 40-50% lower than those in intact women throughout the 50-89 yr age range. Androstenedione levels decreased 27% and SHBG levels increased 30% (P < 0.001) with age in intact, but not oophorectomized, women. Levels of other hormones did not vary with age. Stratification by years since menopause or surgery yielded similar results. These results demonstrate that the postmenopausal ovary remains a critical source of androgen throughout the lifespan of older women. The clinical consequences of lower testosterone levels years after oophorectomy are unknown. Reconsideration of prophylactic oophorectomy and clinical trials to evaluate the effects of androgen replacement after oophorectomy are needed. PMID- 10690871 TI - Adrenal suppression, evaluated by a low dose adrenocorticotropin test, and growth in asthmatic children treated with inhaled steroids. AB - The aim of the present study was to evaluate the prevalence of adrenal suppression and growth retardation in children using moderate doses of budesonide or fluticasone propionate. Seventy-five asthmatic children were randomly divided into three treatment groups: 30 to the fluticasone propionate (FP), 30 to the budesonide (BUD), and 15 to the cromone (CROM) group. FP doses were 500 microg/day during the first 2 months and 200 microg/day thereafter. The respective BUD doses were 800 and 400 microg/day. A low dose ACTH (0.5 microg/1.73 m2) test was performed before treatment and 2, 4, and 6 months later. The test was considered abnormal if the stimulated serum cortisol concentration was more than 2 SD lower than the pretreatment mean (<330 nmol/L). The low dose ACTH test was abnormal after both the high and low steroid doses in 23% of the children. At the 4 month measurement there were more abnormal tests in the BUD (n = 9) than in the FP (n = 5) group (P < 0.05). At that time also the stimulated concentration of serum cortisol was lower in the BUD than in the CROM group (P < 0.01), whereas the difference between the FP and CROM groups was not significant. During the study year the mean decrease in height SD score was 0.23 in the children treated with BUD, 0.03 in the children treated with FP, and 0.09 in the children treated with CROM; the difference between the BUD and FP groups was significant (P < 0.05). In conclusion, the low dose ACTH test revealed mild adrenal suppression in a quarter of the children using moderate doses of inhaled steroids. A FP dose of 200 microg/day caused less adrenal and growth suppression than did a BUD dose of 400 microg/day. PMID- 10690872 TI - Phenotypic features, androgen receptor binding, and mutational analysis in 278 clinical cases reported as androgen insensitivity syndrome. AB - Androgen insensitivity syndrome (AIS) is the most common single entity that results in male under-masculinization, but large cohort studies of AIS have rarely been performed. Over the last decade, nationwide cooperation between pediatric endocrinologists in the United Kingdom has allowed the creation of a database of cases of intersex and ambiguous genitalia where detailed clinical information on every notified case has been collected via a questionnaire. Among the 816 entries recorded by January 1999, there were 105 clinically diagnosed cases of complete AIS (CAIS) and 173 cases of partial AIS (PAIS). A masculinization score was devised by scoring the external phenotype, and a score of 12 represented normal masculinization. Androgen receptor (AR) binding was determined by studying binding capacity (Bmax) and receptor affinity (K(d)), and cases were classified as either zero, abnormal, or normal binding. Mutation screening of all eight exons of the AR gene was performed by single-strand conformational polymorphism analysis, followed by direct DNA sequencing. All cases of PAIS presented within the first month of birth. The median age at presentation of children with CAIS was 1 yr (P10,P90: 0.1,10.4). The testes were palpable in the labioscrotal folds or the inguinal region in 77% and 41% of cases of CAIS and PAIS, respectively. There was marked overlap between the masculinization score of those children with PAIS reared as girls [2.5(P10,P90:1, 6)] and those reared as boys [3(P10,P90:2, 7.5)]. Gonadectomy was performed prepubertally in 66% and postpubertally in 29% of the cases of CAIS. The median age of the latter group was older at 14 yr (P10,P90:0.1,18). No cases of malignancy or carcinoma in situ were reported in the 121 cases of AIS where histology results were available. Biochemical endocrine investigations were reported to have been performed in a greater number of cases of PAIS than CAIS (98% vs. 48%). AR binding was abnormal in 44 of 51 (86%) and 40 of 113 (35%) cases of CAIS and PAIS, respectively. Zero binding was encountered in 29 of 43 (67%) and 1 of 55 (2%) cases of CAIS and PAIS, respectively. Mutational analysis of the AR gene, performed in 102 index cases was positive in 57 of 69 (83%) cases of CAIS and 12 of 43 (28%) cases of PAIS. In 24 of these cases, the mutation identified was novel. The mutations in PAIS cases were all missense, whereas in CAIS the mutations were more diverse. AR binding was only normal in 3 of 69 mutation-positive cases. In the PAIS group, mutation-positive cases had a significantly higher Kd and Bmax compared to the mutation negative cases. The clinical diagnosis of AIS can be confirmed in a significant number of cases by a combination of androgen-binding studies and mutational analysis. There is some correlation between the phenotypic features and the abnormalities discovered on mutational analysis of the AR gene, but there is a need to improve this further by developing optimal bioassays of AR function. The phenotypic heterogeneity among clinically diagnosed cases of AIS emphasizes the need for appropriate comprehensive evaluation of male under-masculinization. PMID- 10690873 TI - Measurement of urinary melatonin: a useful tool for monitoring serum melatonin after its oral administration. AB - The relevance of measuring urinary melatonin (MLT) for human pineal research is sometimes questioned, and the relationship among serum levels of MLT, urinary excretion of the unmetabolized hormone, and excretion of MLTs main metabolite, 6 hydroxymelatonin sulfate (aMT6s), is still uncertain. We applied a well established RIA for measuring MLT in serum to urine samples, characterized its criteria of performance in this body fluid, and used it for human studies. In 16 adolescents, the endogenous overnight MLT secretion, expressed as the area under the concentration time curve, correlated significantly with the amounts of urinary aMT6s (r = 0.86; P < 0.0001) and urinary MLT (r = 0.70; P = 0.0027) excreted during a 16-h observation period. Oral administration of 3 mg exogenous MLT in 17 healthy volunteers resulted in peak MLT serum levels differing 28-fold among subjects (940-27,240 pg/ mL; range). In this study urinary MLT, but not aMT6s, excretion was associated with blood MLT concentrations (r = 0.76; P = 0.0004 vs. r = 0.02; P = 0.93, respectively). Thus, endogenous MLT production can be assessed accurately by measuring either aMT6s or MLT excretion. After oral application of MLT, however, only measurement of MLT excretion is a reliable marker of serum concentrations. Determination of MLT in urine may prove to be a useful tool for drug monitoring after oral administration of the pineal hormone. PMID- 10690874 TI - Diagnosis of thyroid malignant lymphoma by reverse transcription-polymerase chain reaction detecting the monoclonality of immunoglobulin heavy chain messenger ribonucleic acid. AB - Distinguishing between thyroid malignant lymphoma and lymphocytic thyroiditis (Hashimoto's thyroiditis) is quite difficult and problematic. Molecular techniques to detect clonal lymphoid proliferation based on Ig heavy chain (IgH) gene rearrangement may be used to facilitate more accurate diagnosis of malignant lymphoma. We recently established a method for diagnosing thyroid tumors by analyzing ribonucleic acids (RNAs) extracted from the needles used for fine needle aspiration biopsy (aspiration biopsy-RT-PCR). By applying the aspiration biopsy-RT-PCR method to detection of the monoclonality of IgH messenger RNA (mRNA), an accurate molecular-based diagnosis of malignant lymphoma can be established as an adjunct to cytological diagnosis. We first studied RNAs from fresh tissues samples of 8 cases of Hashimoto's thyroiditis and 18 malignant lymphomas to detect the monoclonality of IgH mRNA by seminested RT-PCR. Monoclonality was detected in 8 of 18 (44.4%) malignant lymphomas, but in none of the 8 cases of Hashimoto's thyroiditis. We then studied aspirates from 10 cases of thyroid malignant lymphoma, 4 cases of Hashimoto's thyroiditis, and 1 case each of adenomatous goiter and papillary carcinoma. Monoclonality was detected in the aspirates from 4 of 10 malignant lymphomas (40%), but not from other tissues. Thus, RT-PCR detection of monoclonality of IgH mRNA in addition to cytological examination may be useful in diagnosing thyroid malignant lymphoma. PMID- 10690875 TI - Free androgen index and leptin are the most prominent endocrine predictors of ovarian response during clomiphene citrate induction of ovulation in normogonadotropic oligoamenorrheic infertility. AB - We have previously demonstrated that obese hyperandrogenic amenorrheic women are less likely to ovulate after clomiphene citrate (CC) medication. The present study was designed to identify whether additional endocrine screening characteristics, all potentially involved in ovarian dysfunction in 182 normogonadotropic oligoamenorrheic infertile women, are associated with ovarian response, which may improve overall prediction of CC-resistant anovulation. Standardized endocrine screening took place before initiation of CC medication (50 mg/day; increasing doses up to 150 mg/day if required) from cycle days 3-7. Screening included serum assays for fasting insulin and glucose, insulin-like growth factor I (IGF-I), IGF-binding protein-1 (IGFBP-1), IGFBP-3, free IGF-I, inhibin B, leptin, and vascular endothelial growth factor. Forty-two women (22% of the total group) did not ovulate at the end of follow-up (a total number of 325 cycles were analyzed). Fasting serum insulin, insulin/glucose ratio, IGFBP-1, and leptin were all significantly different in univariate analyses (P < or = 0.02), comparing CC responders vs. nonresponders. Forward stepwise multivariate analyses in combination with factors reported earlier for prediction of patients remaining anovulatory after CC revealed a prediction model including 1) free androgen index (FAI = testosterone/sex hormone-binding globulin ratio), 2) cycle history (oligomenorrhea or amenorrhea), 3) leptin level, and 4) mean ovarian volume. These data suggest that decreased insulin sensitivity, hyperandrogenemia, and obesity, all associated with polycystic ovary syndrome, are prominent factors involved in ovarian dysfunction, preventing these ovaries from responding to stimulation by raised endogenous FSH levels due to CC medication. By using leptin instead of body mass index or waist to hip ratio, the previous model for prediction of patients remaining anovulatory after CC medication could be slightly improved (area under the curve from 0.82-0.85). This may indicate that leptin is more directly involved in ovarian dysfunction in these patients. The capability of insulin and IGFBP-1 to predict patients who remain anovulatory after CC disappears when FAI enters into the model due to a significant correlation between FAI and these endocrine parameters. This suggests that markers for insulin sensitivity (e.g. IGFBP-1 and insulin) are associated with abnormal ovarian function through its correlation with androgens, whereas leptin is directly involved in ovarian dysfunction. PMID- 10690876 TI - Is leptin associated with hypertensive retinopathy? AB - Previous studies have demonstrated that elevated plasma leptin concentrations are associated with essential hypertension. It has also recently been shown that leptin plays a promoting role in angiogenesis, and the vascular endothelium expresses the long form of leptin receptor. Those data led us to hypothesize that leptin might contribute to end-organ damage in hypertension. Thus, in the present study we evaluated the relationship between plasma leptin concentrations and hypertensive retinopathy (HR). One hundred and eleven patients newly diagnosed with essential hypertension [EHT; mean age, 43.5 +/-10.7 yr; body mass index (BMI), 28.1 +/- 4.4 kg/m2; male/female ratio, 71/40] and 79 healthy normotensive control subjects (NT; mean age, 43.6 +/- 9.2 yr; BMI, 28.2 +/- 3.3 kg/m2; male/female ratio, 50/29) were enrolled in the study. For the assessment of retinopathy according to the Keith-Wagener classification, direct and indirect ophthalmoscopy were performed in all subjects after dilatation of the pupils. Plasma leptin levels were significantly higher in EHT (11.8 +/- 11.1 ng/mL) than in NT (7.2 +/- 5.1 ng/mL) (P = 0.003). Plasma leptin concentrations were strongly correlated with BMI in both EHT (r = 0.45; P = 0.001) and NT (r = 0.38; P = 0.001) groups. Plasma leptin in patients with grade 2 HR (24.8 +/- 15.8 ng/mL; n = 22) was significantly higher than that in patients with grade 1 HR (16.1 +/- 4.9 ng/mL; n = 29; P = 0.001), grade 0 HR (5.1 +/- 3.1 ng/mL; n = 60; P = 0.001), and NT (P = 0.001). Plasma leptin in patients with grade 1 HR was also significantly higher than that in patients without retinopathy (P = 0.001) or in NT (P = 0.001). The estimated threshold of plasma leptin concentration for HR was 10.2 ng/mL. This critical leptin level served largely to separate patients with retinopathy from those without retinopathy. In summary, our results show that plasma leptin concentrations increase progressively with higher grades of hypertensive retinopathy even after correction for BMI, suggesting that a critical leptin level is needed for the development of retinopathy. Elevated concentrations of plasma leptin might be secondary to release of leptin by the vascular endothelium damaged by high blood pressure, as an epiphenomenon. However, a pathogenic role for leptin in hypertensive retinopathy cannot be excluded. PMID- 10690877 TI - Association analysis of the cytotoxic T lymphocyte antigen-4 (CTLA-4) and autoimmune regulator-1 (AIRE-1) genes in sporadic autoimmune Addison's disease. AB - Although autoimmune Addison's disease (AAD) may occur as a component of the monogenic autoimmune polyendocrinopathy type 1 syndrome (APS1), it is most commonly found as an isolated disorder or associated with the autoimmune polyendocrinopathy type 2 syndrome (APS2). It is likely that sporadic (non-APS1) AAD is inherited as a complex trait; however, apart from the major histocompatibility complex, the susceptibility genes remain unknown. We have examined polymorphisms at two non-major histocompatibility complex candidate susceptibility loci in sporadic (non-APS1) AAD: the cytotoxic T lymphocyte antigen-4 (CTLA-4) gene and the autoimmune regulator (AIRE-1) gene. DNA samples from AAD subjects (n = 90) and local controls (n = 144 for CTLA-4; n = 576 for AIRE-1) were analyzed for the CTLA-4A/G polymorphism in exon 1 of the CTLA-4 gene and for the common mutant AIRE-1 allele (964de113) in United Kingdom subjects with APS1, by using the restriction enzymes Bst7II and BsrBI, respectively. There was an association of the G allele at CTLA-4A/G in AAD subjects (P = 0.008 vs. controls), which was stronger in subjects with AAD as a component of APS2 than in subjects with isolated AAD. In contrast, the mutant AIRE-1 964del13 allele was carried in one each of the 576 (0.2%) control subjects and the 90 (1.1%) AAD subjects as a heterozygote (P = 0.254, not significant), suggesting that this common AIRE-1 gene abnormality does not have a major role in sporadic (non-APS1) AAD. PMID- 10690878 TI - Disturbed neuroendocrine-immune interactions in chronic fatigue syndrome. AB - The present study was designed to investigate the interaction between neuroendocrine mediators and the immune system in chronic fatigue syndrome (CFS). We examined the sensitivity of the immune system to the glucocorticoid agonist dexamethasone and the beta2-adrenergic agonist terbutaline in 15 adolescent girls with CFS and 14 age- and sex-matched controls. Dexamethasone inhibits T-cell proliferation in healthy controls and in CFS patients. However, the maximal effect of dexamethasone on T-cell proliferation is significantly reduced in CFS patients as compared with controls. The beta2-adrenergic receptor agonist terbutaline inhibits tumor necrosis factor-alpha production and enhances interleukin-10 production by monocytes. Our data demonstrate that the capacity of a beta2-adrenergic agonist to regulate the production of these two cytokines is also reduced in CFS patients. We did not observe differences in baseline or CRH induced cortisol and ACTH between CFS patients and controls. Baseline noradrenaline was similar in CFS and controls, whereas baseline adrenaline levels were significantly higher in CFS patients. We conclude that CFS is accompanied by a relative resistance of the immune system to regulation by the neuroendocrine system. Based on these data, we suggest CFS should be viewed as a disease of deficient neuroendocrine-immune communication. PMID- 10690879 TI - Response of leucine metabolism to hyperinsulinemia in hypothyroid patients before and after thyroxine replacement. AB - We have investigated the effect of hypothyroidism and insulin on protein metabolism in humans. Six hypothyroid patients were studied in a postabsorptive state before and after 5 months of regular treatment for hypothyroidism (153 +/- 17 microg/day of L-T4). The effect of insulin was assessed under hyperinsulinemic euglycemic and eukalemic conditions. Insulin was infused for 140 min at 0.0063 +/ 0.0002 nmol/kg x min. An amino acid infusion was used to blunt insulin-induced hypoaminoacidemia. Whole body protein turnover was measured using L-[1-13C] leucine. When compared to L-T4-induced subclinical thyrotoxic state, hypothyroidism induced a significant decrease (P < 0.05) in leucine endogenous appearance rate (a reflection of proteolysis; 0.89 +/- 0.09 vs. 1.33 +/- 0.05 micromol/kg x min), oxidation (0.19 +/- 0.02 vs. 0.25 +/- 0.03 micromol/kg x min), and nonoxidative disposal (a reflection of protein synthesis; 0.87 +/- 0.11 vs. 1.30 +/- 0.05 micromol/ kg x min). Insulin lowered proteolysis during both the subclinical thyrotoxic and hypothyroid states. Hypothyroidism impaired the antiproteolytic effects of insulin. Thyroid hormones are, therefore, essential for the normal antiproteolytic action of insulin. PMID- 10690880 TI - Isolated familial somatotropinomas: establishment of linkage to chromosome 11q13.1-11q13.3 and evidence for a potential second locus at chromosome 2p16-12. AB - The majority of somatotropinomas are sporadic, although a small number occur with a familial aggregation, either as a component of an endocrine neoplasia complex that includes multiple endocrine neoplasia type 1 (MEN-1) and Carney complex (CNC) or as isolated familial somatotropinomas (IFS). IFS is defined as the occurrence of at least two cases of acromegaly or gigantism in a family that does not exhibit MEN-1 or CNC. This rare disease is associated with loss of heterozygosity (LOH) on chromosome 11q13, the locus of the MEN-1 gene, although the MEN-1 sequence and expression appear normal. These data suggest the presence of another tumor suppressor gene located at 11q13 that is important in the control of somatotrope proliferation. To establish linkage of IFS to 11q13 and to define the candidate interval of the IFS gene, we performed haplotype and allelotype analyses on two families with IFS. Collectively, allelic retention in one tumor and a recombinant haplotype in an affected individual mapped the tumor suppressor gene involved in the pathogenesis of IFS to a region of 8.6 cM between polymorphic microsatellite markers D11S1335 and INT-2 located at chromosome 11q13.1-13.3. Maximum two-point LOD scores for five markers within this region were 3.0 or more at theta = 0.0. As somatotropinomas are the predominant pituitary tumor subtype associated with CNC and arise before 30 yr of age, which is strikingly similar to the age at diagnosis for IFS, we explored the possibility that the putative CNC genes might also contribute to the pathogenesis of IFS. Although the genetic defect responsible for the complex is unknown, CNC has been mapped by linkage analysis to chromosomes 2p15-16 and 17q23-24 in different kindreds. Two-point LOD scores less than -2.0 were obtained using marker D17S949 from chromosome 17q23-24, excluding linkage. However, LOD scores of 2.5 were obtained for markers within 2p16-12; therefore, linkage of IFS to chromosome 2p cannot be excluded. This report establishes linkage of the tumor suppressor gene involved in the pathogenesis of IFS to chromosome 11q13.1-13.3 and identifies a potential second locus at chromosome 2p16-12. PMID- 10690881 TI - Normal reproductive function in leptin-deficient patients with lipoatropic diabetes. AB - To further examine the relationships between leptin and female reproductive axis, we conducted hormonal studies in two patients with lipoatropic diabetes that occurred before puberty. Despite complete atrophy of sc and visceral adipose tissue, menarche occurred in these two patients between 11-12 yr of age, followed by regular menstrual cycles. One patient had been pregnant three times, giving birth to children who did not develop the disease. In our two patients, repeated analysis revealed leptin levels below 1 ng/mL (normal range for 20 insulin treated diabetic women, 2-23 ng/mL for body mass index of 14-39 kg/m2; personal data). We measured peripheral levels of estradiol, progesterone, FSH, LH, free testosterone, and androstenedione within the first 5 days of the menstrual cycle, and we tested the reactivity of pituitary after iv injection of 100 microg GnRH. The variation in body temperature in the morning before arising was also analyzed. We showed that 1) all measured levels of hormones were in the normal range for both patients; and 2) low levels of leptin did not impair the development of reproductive function in one patient and was associated with normal gonadal function in both patients. We conclude that puberty and fertility can occur despite chronic low serum levels of leptin. This suggests that leptin is not fundamental to the maintenance of normal reproductive function in humans. PMID- 10690882 TI - Alendronate and estrogen effects in postmenopausal women with low bone mineral density. Alendronate/Estrogen Study Group. AB - The bisphosphonate alendronate and conjugated equine estrogens are both widely used for the treatment of postmenopausal osteoporosis. Acting by different mechanisms, these two agents decrease bone resorption and thereby increase or preserve bone mineral density (BMD). The comparative and combined effects of these medications have not been rigorously studied. This prospective, double blind, placebo-controlled, randomized clinical trial examined the effects of oral alendronate and conjugated estrogen, in combination and separately, on BMD, biochemical markers of bone turnover, safety, and tolerability in 425 hysterectomized postmenopausal women with low bone mass. In addition, bone biopsy with histomorphometry was performed in a subset of subjects. Treatment included placebo, alendronate (10 mg daily), conjugated equine estrogen (CEE; 0.625 mg daily), or alendronate (10 mg daily) plus CEE (0.625 mg daily) for 2 yr. All of the women received a supplement of 500 mg calcium daily. At 2 yr, placebo-treated patients showed a mean 0.6% loss in lumbar spine BMD, compared with mean increases in women receiving alendronate, CEE, and alendronate plus CEE of 6.0% (P < 0.001 vs. placebo), 6.0% (P < 0.001 vs. placebo), and 8.3% (P < 0.001 vs. placebo and CEE; P = 0.022 vs. alendronate), respectively. The corresponding changes in total proximal femur bone mineral density were +4.0%, +3.4%, +4.7%, and +0.3% for the alendronate, estrogen, alendronate plus estrogen, and placebo groups, respectively. Both alendronate and CEE significantly decreased biochemical markers of bone turnover, specifically urinary N-telopeptide of type I collagen and serum bone-specific alkaline phosphatase. The alendronate plus CEE combination produced slightly greater decreases in these markers than either treatment alone, but the mean absolute values remained within the normal premenopausal range. Alendronate, alone or in combination with CEE, was well tolerated. In the subset of patients who underwent bone biopsies, histomorphometry showed normal bone histology with the expected decrease in bone turnover, which was somewhat more pronounced in the combination group. Thus, alendronate and estrogen produced favorable effects on BMD. Combined use of alendronate and estrogen produced somewhat larger increases in BMD than either agent alone and was well tolerated. PMID- 10690883 TI - Behavioral effects of prenatal versus postnatal androgen excess in children with 21-hydroxylase-deficient congenital adrenal hyperplasia. AB - Systematic behavioral studies show that females with congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH) are masculinized and defeminized in several ways; compared to their sisters, they play more with boys' toys, are more likely to use aggression when provoked, and show less interest in infants. We studied the extent to which these behavioral changes could be attributed to high levels of androgens in the prenatal vs. postnatal periods in 23 girls with CAH, aged 3-12 yr. Sex-atypical behavior was significantly associated with degree of inferred prenatal, but not postnatal, androgen excess; marked boy-typical play was associated with severe salt-wasting CAH, early age at diagnosis, and moderate genital masculinization at birth, but not with bone age advance, concurrent or cumulative high levels of 17-hydroxyprogesterone, or accelerated growth velocity in early childhood. Aggression and interest in infants were not consistently associated with indicators of prenatal or postnatal androgen excess, probably because those behaviors were measured less reliably than was toy play. The results are consistent with the idea that behavioral masculinization in girls with CAH results from high levels of androgens during fetal development and not in postnatal life. PMID- 10690884 TI - Interleukin-1 receptor antagonist ribonucleic acid and protein expression by cultured Graves' and normal orbital fibroblasts is differentially modulated by dexamethasone and irradiation. AB - Recent data have indicated that orbital fibroblasts (OF) can be stimulated to produce marked quantities of interleukin-1 receptor antagonist (IL-1RA), a powerful inhibitor of the proinflammatory activities of interleukin-1 in the orbital tissues in Graves' ophthalmopathy (GO). We examined whether the beneficial effects of dexamethasone or irradiation, the two main therapeutic modalities applied in patients with active GO, may be related to their capacity to alter IL-1RA ribonucleic acid (RNA) and protein expression in OF. Early passages of cultured OF were obtained from orbital connective tissue and extraocular muscle of patients with severe active GO and five control subjects. Modulation of the two variants of IL-1RA, intracellular IL-1RA (icIL-1RA) and soluble IL-1RA (sIL-1RA), was studied after exposure of OF to increasing concentrations of dexamethasone (10(-10)-(10(-6) mol/L)), the glucocorticoid receptor antagonist RU 38486 (10(-3) mol/L), or combinations thereof. Alternatively, cell monolayers were exposed to increasing doses of UV irradiation (0.1-1 J/cm2) or ionizing irradiation (0.2-2 Gy). The IL-1RA gene and protein variants were analyzed by RT-PCR, immunocytochemistry, immunoblotting, and enzyme linked immunosorbent assay. Dexamethasone inhibited IL-1RA RNA steady state levels in GO OF and control OF in a dose-dependent manner. Combined exposure of OF to dexamethasone and RU 38486 completely restored baseline levels of IL-1RA RNA. By contrast, low doses of UV and ionizing irradiation dose dependently up regulated IL-1RA-specific transcripts in GO OF and control OF, whereas higher doses were less effective. Immunoblotting and enzyme-linked immunosorbent assay revealed suppression of IL-1RA immunoreactivity after treatment with dexamethasone and enhanced expression of IL-1RA by GO OF and normal OF after low doses of UV and ionizing irradiation. Our results indicate that, in contrast to dexamethasone, low doses of irradiation stimulate expression of the IL-1RA gene and protein variants in OF. Induction by irradiation of IL-1RA expression in target cells of the orbital immune process represents an as yet unrecognized mechanism by which orbital radiotherapy may exert some of its beneficial therapeutic effects in patients with active GO. PMID- 10690885 TI - Asp361Val Mutant of alkaline phosphatase found in patients with dominantly inherited hypophosphatasia inhibits the activity of the wild-type enzyme. AB - Hypophosphatasia is characterized by the hypomineralization of bone associated with the mutation of the tissue-nonspecific alkaline phosphatase (TNSALP) gene. Although the disease is usually autosomal recessive, an autosomal dominant form is also recognized. Approximately 50 mutations have been found in the TNSALP gene in patients with hypophosphatasia. However, the mutations identified to date do not seem to account for the dominantly inherited form of the disease. We have examined a German family in which the father and all 4 children were affected with hypophosphatasia, whereas the mother was healthy. The affected members of this family showed premature loss of deciduous teeth at or shortly before 2 yr of age and low levels of serum ALP with elevated levels of urinary phosphoethanolamine. DNA analysis by direct sequencing revealed a heterozygous missense mutation that caused the conversion of amino acid Asp to Val at position 361 (D361V) in the patients. Another substitution was detected in exon 12 (Val to Ala conversion at codon 505: V505A) in 1 allele of the mother and 3 children, indicating no association of the substitution with the disease. Reconstruction experiments demonstrated that the D361V mutant protein lost its enzymatic activity and that it inhibited the function of wild-type enzyme when coexpressed in COS-7 cells. On the other hand, the V505A mutant exhibited enzymatic activities equal to those of the wild-type ALP. It is likely that the mutant D361V protein forms dimers with the wild-type protein, and the protein-protein interaction contributes to the dominant effect of the mutant D361V. The mutation that causes D361V is the first one proven to be associated with the dominant form of hypophosphatasia. PMID- 10690886 TI - Intramuscular glycogen and intramyocellular lipid utilization during prolonged exercise and recovery in man: a 13C and 1H nuclear magnetic resonance spectroscopy study. AB - Depletion of muscle glycogen is considered a limiting performance factor during prolonged exercise, whereas the role of the intramyocellular lipid (IMCL) pool is not yet fully understood. We examined 1) intramyocellular glycogen and lipid utilization during prolonged exercise, 2) resynthesis of muscle glycogen and lipids during recovery, and 3) changes in glycogen content between nonexercising and exercising muscles during recovery. Subjects ran on a treadmill at submaximal intensity until exhaustion. Glycogen concentrations were assessed in thigh, calf, and nonexercising forearm muscle, and IMCL content was measured in soleus muscle using magnetic resonance spectroscopy techniques. At the time of exhaustion, glycogen depletion was 2-fold greater in calf than in thigh muscles, but a significant amount of glycogen was left in both leg muscles. The glycogen concentration in nonexercising forearm muscle decreased during the initial 5 h of recovery to 73% of the baseline value. Duringthe exercise, the IMCL content decreased to 67% and subsequently during recovery increased to 83% of the baseline value. In summary, we found during prolonged running 1) significantly greater muscle glycogen utilization in the calf muscle group than in the thigh muscle group, 2) significant utilization of IMCL in the soleus muscle, and 3) a decrease in glycogen content in nonexercising muscle and an increase in glycogen content in recovering muscles during the postexercise phase. These latter data are consistent with the hypothesis that there is transfer of glycogen by the glucose-lactate and the glucose-->alanine cycle from the resting muscle (forearm) to recovering muscles (thigh and calf) after running exercise. PMID- 10690887 TI - Processing of procorticotropin-releasing hormone (pro-CRH): molecular forms of CRH in normal and preeclamptic pregnancy. AB - This study examined the different molecular forms of CRH in normal and preeclampsia maternal plasma and protease-blocked placental extracts using antibodies to different regions of the CRH precursor, pro-CRH. In the absence of protease inhibitors, chromatographed normal placental extracts contained four peaks of immunoreactivity corresponding to unprocessed approximately 19-kDa pro CRH, its approximately 8-kDa intermediate metabolite, pro-CRH125-194, its approximately 2.8-kDa midportion fragment, pro-CRH125-151, and 4.75-kDa CRH1-41. However, if protease inhibitors were included in the extraction medium, only pro CRH and pro-CRH125-194 were found. Pro-CRH processing was more extensive in protease-blocked preeclampsia placentas than in those from normal pregnancy, with three peaks corresponding to pro-CRH, proCRH125-194, and mature CRH1-41 peptide found. Using quantitative competitive PCR, the messenger ribonucleic acid levels of CRH precursor in preeclampsia placentas were 1.7-fold higher than those in normal placentas (37.83 +/- 3.48 vs. 21.83 +/- 2.59 attomoles/microg total ribonucleic acid, respectively; P < 0.005). Preeclampsia placentas contained significantly more CRH1-41 cross-reactivity (4.72 +/- 1.22 pmol/g) than normal term placentas (1.52 +/- 0.39 pmol/g; P < 0.048) extracted in medium containing protease inhibitors. The content of pro-CRH(125+/-151)-reactive species in these extracts followed the same pattern, with more immunoreactivity detected in preeclampsia placentas (4.23 +/- 1.39 pmol/g) than in those from normal term pregnancies (1.44 +/- 0.32 pmol/g; P < 0.01). Sequential plasma samples from 10 women with normal pregnancy and 5 women with preeclampsia were assayed for pro CRH(125-151)- and CRH(1-41)-immunoreactive species In normal pregnancy, maternal plasma CRH(1-41) immunoreactivity rose with increasing gestational age, reaching 460 +/- 48 pmol/L at term. In women with preeclampsia, CRH(1-41) levels at each gestational age point were higher than those at the equivalent stage of normal pregnancy. In contrast, the levels of pro-CRH(125-151)-immunoreactive species remained barely detectable throughout normal and preeclamptic pregnancy. Both pro CRH and CRH(1-41), but not pro-CRH(125-151), were shown to bind to the plasma CRH binding protein. Our findings highlight the importance of protection of placental tissue from degrading enzymes during extraction and show that most of the CRH in the human placenta exists as unprocessed pro-CRH, with very little in the form of CRH(1-41) except in preeclampsia. Our studies using maternal plasma indicate that CRH(1-41) is the only one of the pro-CRH fragments studied to be maintained in significant amounts in the maternal circulation and also the only fragment studied for which a specific plasma binding protein exists. PMID- 10690888 TI - Expression of oncofetal fibronectin messenger ribonucleic acid in fibroblasts in the thyroid: a possible cause of false positive results in molecular-based diagnosis of thyroid carcinomas. AB - Oncofetal fibronectin (onfFN) messenger ribonucleic acid (mRNA) is abundantly expressed in thyroid papillary and anaplastic carcinomas. These carcinomas can be preoperatively diagnosed by the detection of onfFN mRNA in fine needle aspiration biopsies (FNABs). However, previous reports have noted that the expression of onfFN mRNA was observed in 3.7% of the FNABs that were diagnosed as negative cytology. To clarify this discrepancy, we examined the expression of onfFN mRNA in fibroblasts in the thyroid. By RT-PCR and real-time quantitative RT-PCR analyses, we detected a high copy number of onfFN mRNA in cultured fibroblasts obtained from the normal thyroid tissues dissected surgically. Thus, we conclude that the contaminated fibroblasts in FNABs due to tumor necrosis or acute or chronic inflammation may be a cause of false positive results in molecular-based diagnosis of thyroid carcinomas. PMID- 10690889 TI - Carbohydrate metabolism during long-term growth hormone (GH) treatment and after discontinuation of GH treatment in girls with Turner syndrome participating in a randomized dose-response study. Dutch Advisory Group on Growth Hormone. AB - To assess possible side-effects of GH treatment with supraphysiological doses on carbohydrate (CH) metabolism in girls with Turner syndrome (TS) during long term GH treatment and after discontinuation of GH treatment, the results of oral glucose tolerance tests and hemoglobin A1c measurements were analyzed in 68 girls with TS participating in a randomized dose-response trial. These previously untreated girls, aged 2-11 yr, were randomly assigned to 1 of 3 GH dosage groups: group A, 4 IU/m2 x day (-0.045 mg/kg x day); group B, first year ,4 IU/m2 day; thereafter, 6 IU/m2 x day (approximately 0.0675 mg/kg x day); group C, first year, 4 IU/m2 x day; second year, 6 IU/m2 x day; thereafter, 8 IU/m2 x day (approximately 0.090 mg/kg x day). After the first 4 yr, girls 12 yr of age or older started with 5 microg/kg BW-day 17beta-estradiol for induction of puberty. To assess the effects of long term high dose GH treatment on CH metabolism, the 7 yr data from the oral glucose tolerance tests in 9 girls of group C were evaluated (group C1). To determine whether the changes in CH metabolism during GH treatment would persist after discontinuation of GH treatment, the data for 28 girls who had reached adult height (group A, n = 9; group B, n = 10; group C, n = 9) were evaluated at baseline, after 4 yr of GH treatment, and 6 months after discontinuation of GH. Seven-year data for group C1 showed that glucose levels did not significantly change during GH treatment, whereas fasting insulin levels as well as glucose-induced insulin levels increased significantly. The data for the 28 girls who were treated with GH for a mean (SD) period of 85.3 (13.3) months demonstrated that the GH-induced higher insulin levels decreased to values close to or equal to pretreatment values after discontinuation of GH treatment. Changes in CH variables were not significantly related to the GH dose. Hemoglobin A1c levels never showed an abnormal value. The prevalence of impaired glucose tolerance was low, and none of the girls developed diabetes mellitus. In conclusion, long term GH treatment with dosages up to 8 IU/m2 x day in girls with TS has no adverse effects on glucose levels, but induced higher levels of insulin, indicating relative insulin resistance. The increased insulin levels during long term GH treatment decreased after discontinuation of GH treatment to values close to or equal to pretreatment values. Although the reversibility of the effects of long term GH is reassuring, the consequence of long term hyperinsulinism is still unknown. PMID- 10690890 TI - Relationship between disease duration and predominant orbital T cell subset in Graves' ophthalmopathy. AB - We sought to determine whether the predominant orbital T helper (T(H)) cell subset in orbital T cell clones established from patients with Graves' ophthalmopathy (GO) might be related to disease duration. A total of 117 clones were established from orbital adipose/connective tissues of 6 GO patients, and cytokine production was measured in 57 CD3+CD4+ clones. T(H)1-type clones were predominant in cultures from patients with recent onset (<2 yr) Graves' hyperthyroidism (n = 44; TH1/TH0/TH2 = 57/29/14%) or GO (n = 53 clones; TH1/TH0/TH2 = 47/30/23%). In contrast, TH2-type clones predominated in cultures from patients with more remote onset (>2 yr) hyperthyroidism (n = 13; TH1/TH0/TH2 = 0/31/69%; P < 0.005) or GO (n = 4; TH1/TH0/TH2 = 0/25/75%; P = 0.05). In addition, we established T cell clones from 1 TH1-dominant patient with recent onset thyroid and eye disease using either IL-2 (12.5 ng/mL) alone or IL-2 plus IL-4 (5 ng/mL) and found no shift toward recovery of TH2-type clones in the latter. In conclusion, although the CD3+CD4+ clones characterized were not necessarily tissue antigen specific, our findings suggest that cell-mediated (TH1 type) immune reactions may predominate in the orbit in early GO, whereas humoral immunity (TH2-type) might play the greater role in later stages of the disease. PMID- 10690891 TI - Human somatostatin receptor subtypes in acromegaly: distinct patterns of messenger ribonucleic acid expression and hormone suppression identify different tumoral phenotypes. AB - Recently, studies using somatostatin (SRIF) analogs preferential for either the SRIF receptor 2 (SSTR2) or the SSTR5 subtype demonstrated a variable suppression of GH and PRL release from GH-secreting human adenomas. These data suggested the concept of SSTR subtype specificity in such tumors. In the present study the quantitative expression of messenger ribonucleic acid (mRNA) for the 5 SSTR subtypes and the inhibitory effects of SRIF14; SRIF28; octreotide; the SSTR2 preferential analog, BIM-23197; and the SSTR5-preferential analog, BIM-23268, on GH and PRL secretion were analyzed in cells cultured from 15 acromegalic tumors. RT-PCR analysis revealed a consistent pattern of SSTR2 and SSTR5 mRNA expression. SSTR5 mRNA was expressed at a higher level (1052 +/- 405 pg/pg glyceraldehyde-3 phosphate dehydrogenase) than SSTR2 mRNA (100 +/- 30 pg/pg glyceraldehyde-3 phosphate dehydrogenase). However, only SSTR2 mRNA expression correlated with the degree of GH inhibition induced by SRIF14, SRIF28, and BIM-23197. The SSTR5 preferential compound inhibited GH release in only 7 of 15 cases. In cells cultured from the 10 mixed adenomas that secreted both GH and PRL, RT-PCR analysis revealed a consistent coexpression of SSTR5, SSTR2, and SSTR1 mRNA. In all cases SRIF14, SRIF28, and the SSTR5-preferential analog, BIM-23268, significantly suppressed PRL secretion, with a mean maximal inhibition of 48 +/- 4%. In contrast, the SSTR2-preferential analogs, BIM-23197 and octreotide, were effective in suppressing PRL in only 6 of 10 cases. In cells cultured from adenomas taken from patients partially responsive to the SRIF analog, octreotide, partial additivity in suppressing both GH and PRL secretion was observed when the SSTR2- and SSTR5-preferring analogs, BIM-23197 and BIM-23268, were tested in combination. Our data show a highly variable ratio of the SSTR2 and SSTR5 transcripts, according to tumors. The SSTR2-preferring compound consistently inhibits GH release, whereas the SSTR5-preferring compound is the main inhibitor of PRL secretion. When both drugs are combined, the partial additivity observed in mixed GH- plus PRL-secreting adenomas may be of interest in the therapeutic approach of such tumors. PMID- 10690892 TI - Plasma free fatty acids and endothelium-dependent vasodilation: effect of chain length and cyclooxygenase inhibition. AB - Free fatty acids (FFA) are known to interfere with glucose metabolism. Moreover, it has been shown that they are able to impair the endothelium-dependent vasodilation. Therefore, we sought to determine whether their negative effect on endothelial function depends on their chain length or on their ability to modify PG production. Fourteen normal volunteers were studied under baseline conditions and then randomly allocated to two of the following four studies: 1) long chain triglyceride (LCT) emulsion and heparin infusion (n = 7), 2) infusion of an emulsion containing 56% medium chain triglycerides (MCT) and 44% LCT plus heparin (n = 7), 3) infusion of LCT and heparin preceded by an i.v. bolus of 900 mg lysine-salicylate (ASA; n = 7), and 4) after an i.v. bolus of ASA (n = 7). Basal forearm blood flow (FBF), endothelium-dependent vasodilation in response to intraarterial acetylcholine (Ach), and endothelium-independent vasodilation in response to intraarterial nitroprusside were assessed by venous occlusion plethysmography. Both LCT and MCT infusions significantly increased basal FBF from 1.58 +/- 0.35 to 2.60 +/- 0.76 and 2.28 +/- 0.56 mL/min 100 mL tissue, respectively (both P < 0.05). This increase was also observed for LCT plus heparin, but not after ASA alone. The percent increase in FBF during Ach was lowered during both LCT (252 +/- 34% of the ratio infused/control arm at maximal Ach dose) and MCT (255 +/- 41%) compared to the baseline conditions (436 +/- 44%; both P < 0.05). The response to Ach was also lower during LCT plus ASA, whereas it was similar to baseline with ASA alone. No differences were observed in the response to nitroprusside among the experimental conditions. In conclusion, 1) the effect of FFA on endothelium-dependent vasodilation is independent of their chain length; 2) both LCT and MCT increase baseline FBF, independently from cyclooxygenase inhibition; and 3) acute ASA administration does not affect endothelium-dependent vasodilation. The FFA effect on the endothelial response to Ach may contribute to altered endothelial function and, hence, to the development and progression of atherosclerotic cardiovascular disease. PMID- 10690893 TI - Effects of cortisol and growth hormone on lipolysis in human adipose tissue. AB - The in vitro effects of cortisol and GH on basal and stimulated lipolysis in human adipose tissue were studied using a tissue incubation technique. After preincubation for 3 days in control medium containing insulin, adipose tissue pieces were exposed to cortisol for 3 days. GH was added to the cortisol containing medium during the last 24 h (day 6). Adipocytes were then isolated, and lipolysis was studied in the absence and presence of isoprenaline, noradrenaline, forskolin, and N-6-monobutyryl-cAMP. Cortisol reduced the basal rate of lipolysis (P < 0.01) and the sensitivity to isoprenaline compared to the control values (P < 0.01). Addition of GH to the cortisol-containing medium increased the basal rate of lipolysis (P < 0.01) and the sensitivity to isoprenaline (P < 0.01) to the control level and increased the maximum isoprenaline-induced lipolytic activity (P < 0.01). Similar effects were obtained in the presence of noradrenaline. Maximum forskolin-induced lipolytic activity was reduced after exposure of the tissue to cortisol (P < 0.05), whereas addition of GH antagonized this effect (P < 0.01). Induction of the maximum lipolytic activity with N-6-monobutyryl-cAMP was not influenced by the preceding hormone exposure. Addition of GH alone during the last 24 h of incubation increased the basal rate of lipolysis (P < 0.05) and resulted in a borderline significant increase in the maximum isoprenaline-induced lipolytic activity (P = 0.055), suggesting that GH induces lipolysis also in the absence of glucocorticoids. Thus, cortisol and GH have opposite effects on the basal lipolytic activity in human adipose tissue in vitro as well as on the sensitivity to catecholamines, GH being the lipolytic and cortisol the antilipolytic agent. The present findings are in agreement with in vivo observations. PMID- 10690894 TI - Leptin, nutrition, and reproduction: timing is everything. PMID- 10690895 TI - Circulating concentrations of nocturnal leptin, growth hormone, and insulin-like growth factor-I increase before the onset of puberty in agonadal male monkeys: potential signals for the initiation of puberty. AB - The factor(s) responsible for initiating the developmental increase in nocturnal gonadotropin-releasing hormone secretion, defining the onset of puberty, are not known. Although signals regulating prepubertal growth seem to be obvious candidates to control such a process, it is unclear whether prepubertal alterations occur in these growth-related factors such that they might provide the brain information on changing body size. Using samples analyzed previously describing the initiation of nocturnal pulsatile LH secretion in agonadal male monkeys (Endocrinology 139: 2774-2783, 1998), developmental changes in plasma concentrations of leptin, GH, and insulin-like growth factor I (IGF-I) were determined to test the hypothesis that an increase in circulating levels of one or all of these growth-derived signals precedes the onset of puberty. Hormone concentrations were determined in five juvenile males at 10-day intervals from approximately 60 days before and 50 days after the initiation of pulsatile nocturnal LH secretion. Leptin concentrations were determined in samples obtained at 1000 and 2200 h, 36 and 48 h before the nocturnal assessment of pulsatile LH. Mean nocturnal GH concentrations were determined from the sequential samples collected at night. IGF-I was determined in the 1000- or 2200-h presequential samples. Although daytime leptin concentrations did not increase developmentally, nocturnal leptin levels increased significantly during the 30 days before the onset of puberty. Furthermore, both nocturnal GH and IGF-I concentrations showed a significant sustained increase from the early prepubertal period to the 30 days preceding the onset of puberty. These data are the first to demonstrate an increase in nocturnal leptin and GH-induced IGF-I secretion prior to the onset of puberty in the agonadal male monkey and that these developmental changes occur independent of the gonadal influences. These findings provide justification for empirical investigation of the role of leptin and the GH axis, in particular IGF I, in regulating developmental increases in pulsatile nocturnal gonadotropin releasing hormone secretion initiating puberty in primates. PMID- 10690896 TI - The skin produces urocortin. AB - Since the skin produces POMC peptides, in the present work we investigated local production of urocortin, a peptide related to CRH, the normal endogenous stimulant for POMC. Urocortin immunoreactivity was detected by direct RIA in extracts of human skin, mouse skin (C57BL-6 strain), cultured cells from established lines of human melanoma and squamous cell carcinoma, human keratinocytes (Ha-CaT), and hamster melanomas. Addition of a reverse phase high performance liquid chromatography step before the RIA confirmed the presence of urocortin, as the immunoreactivity eluted at the same retention time as urocortin standard in extracts from HaCaT keratinocytes and mouse skin. Using the tandem technique of liquid chromatography-mass spectrometry, we identified a peptide with the same mass and retention time as the urocortin standard in human skin extracts. The urocortin antigen could be immunolocalized to normal keratinocytes of the epidermis and hair follicle, epithelium of sweat and sebaceous glands, dermal skeletal muscle, and nevocytes; it was also detected in melanoma and basal cell carcinoma cells. RT-PCR amplification of ribonucleic acid from human skin, cultured keratinocytes, and melanoma cells showed a 145-kb fragment from the coding region of exon 2 of the urocortin gene in all of the tested sources. Lastly, sequencing of the amplified fragment confirmed 100% homology with the known sequence of the urocortin gene. In conclusion, we now demonstrate that human skin and mouse skin as well as cultured keratinocytes and melanoma cells exhibit functional expression of the urocortin gene with actual production of urocortin peptide. PMID- 10690897 TI - Tumor necrosis factor-alpha promotes proliferation of endometriotic stromal cells by inducing interleukin-8 gene and protein expression. AB - Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. We and others showed that several cytokine levels, including interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNFalpha), are elevated in the peritoneal fluid of women with endometriosis compared with those in women without endometriosis. We also demonstrated that the addition of IL-8 to the culture medium stimulated the proliferation of cultured endometriotic stromal cells. TNFalpha is a multipotent cytokine that induces IL-8 production in various cell types. Therefore, we hypothesized that TNFalpha may also contribute to the pathogenesis of endometriosis by inducing the production of IL-8. To test this hypothesis, we analyzed the peritoneal fluid concentrations of IL-8 and TNFalpha using enzyme linked immunosorbent assay (ELISA). We observed a significant correlation between the levels of TNFalpha and IL-8 in the peritoneal fluid of endometriosis patients. We also obtained the endometriotic stromal cells from chocolate cyst linings of the ovary. The expression of the receptors for TNFalpha (TNFR) was examined by RT-PCR. We observed the expression of both TNFR-I and TNFR-II genes in endometriotic stromal cells. The expression of IL-8 gene and protein was analyzed by Northern blot hybridization and enzyme-linked immunosorbent assay, respectively. TNFalpha induced the gene and protein expression of IL-8 in endometriotic stromal cells in a dose-dependent fashion. The addition of TNFalpha promoted the proliferation of the endometriotic stromal cells, and the stimulatory effects of TNFalpha were abolished by adding anti-IL-8 antibody. We demonstrated for the first time that TNFalpha stimulated proliferation of endometriotic stromal cells through induction of IL-8 gene and protein expression. We concluded that the TNFalpha may be one of the essential factors for the pathogenesis of endometriosis. PMID- 10690898 TI - Interferon-gamma induces interleukin-1 converting enzyme expression in pancreatic islets by an interferon regulatory factor-1-dependent mechanism. AB - Whereas nitric oxide (NO) production is associated with the toxic effect of cytokines on rodent pancreatic beta-cells, cytokine-induced apoptosis in human islets may occur independently of NO. The cysteine protease interleukin (IL)-1 converting enzyme (ICE) is a key proapoptotic caspase. Our aim was therefore to analyze the effect of cytokines on ICE expression in human, rat, and mouse islets and rat insulinoma cells. ICE messenger RNA (mRNA) expression was highly up regulated after 6-, 24-, and 72-h exposure of human islets to interferon (IFN)gamma, tumor necrosis factor (TNF)alpha + IFNgamma or IL-1beta + TNFalpha + IFNgamma, paralleled by increased iNOS (the inducible form of NO synthase) expression and NO production after exposure to the combined cytokines but not to IFNgamma or TNFalpha + IFNgamma. Cytokine-induced NO-independent ICE transcription was confirmed using iNOS inhibitors. Exposure of rat and mouse islets, or rat insulinoma cells, for 24 h to IFNgamma alone or in combination with the two other cytokines also resulted in a highly significant ICE mRNA expression. ICE transcription was not inducible in islets from IFN regulatory factor-1 knock-out mice, suggesting a key-role of this transcription-factor in cytokine-mediated ICE expression in pancreatic islets. In conclusion, cytokines and IFNgamma in particular increase ICE mRNA expression in pancreatic islet cells and beta-cell lines, independently of NO synthesis, suggesting that ICE up regulation may be involved in cytokine-induced NO-independent apoptosis of human islets. PMID- 10690899 TI - Regulated CYP19 aromatase transcription in breast stromal fibroblasts. AB - Extraglandular estrogen synthesis mediates the proliferation of estrogen responsive breast cancer in postmenopausal women. Aromatase, the cytochrome P450 Cyp19 enzyme, catalyzes the rate-limiting step in estrogen biosynthesis. Activity is present in both normal and neoplastic breast tissue, and Cyp19 protein is localized by immunohistochemistry predominantly in breast stromal fibroblasts. In cultured breast stromal fibroblasts, both activity and Cyp19 messenger ribonucleic acid are increased to a substantial degree by hormonal and growth factor regulators of transcription. Transcriptional regulation of CYP19 is complex in breast tissues, in which exon switching in the usage of alternative first exons occurs from predominantly EI.4 in breast tissue from cancer-free women to predominantly EI.3 and PII in breast tumors and quadrants with or without tumor. The present study questioned whether the first exon switch occurs as a result of an inherent difference between fibroblasts in normal and tumor tissues or because of differences in local regulators between these tissues. To distinguish between these two possibilities, we examined fibroblasts cultured from breast tumor, benign breast, and reduction mammoplasty tissues for the ability of various CYP19 transcriptional regulators to modulate first exon EI.3, EI.4, and PII usage. A semiquantitative RT-PCR method was used to identify transcripts containing six of the nine known CYP19 first exons. Combinations of cAMP and Dex regulated transcription from first exons EI.3, EI.4, and PII in fibroblasts cultured from all tissues, but not in reduction mammoplasty epithelial cells. These results provide evidence that the fibroblasts from these breast tissues are not inherently different in transcriptional regulation of CYP19 first exon usage and that transcriptional regulatory molecules are likely to mediate the exon switch phenomenon. PMID- 10690900 TI - Androgen influences transforming growth factor-beta1 gene expression in human adrenocortical cells. AB - Sex steroid hormones have been shown to affect adrenocortical function and trophism, yet little is known about androgen action in human adrenocortical gland. In this study we examined the effects of androgens on transforming growth factor-beta1 (TGF/beta1) production by the human adrenocortical cell line, NCI H295, which we recently demonstrated to express androgen receptor and whose growth is significantly reduced by dihydrotestosterone (DHT) treatment. TGFbeta1 is an important regulator of human adrenal development, with marked effects on steroid-producing cell function, and the production of distinct TGFbeta subtypes has been suggested to be regulated by steroid hormones in several tissues. To address potential TGFbeta1 induction by DHT, quantitative PCR and enzyme-linked immunoadsorbent assay were performed in NCI-H295 cells treated with DHT (from 10( 12)-10(-9) mol/L). DHT led to a significant dose-dependent increase in TGFbeta1 messenger ribonucleic acid expression and in biologically active TGFbeta1 protein levels in the conditioned media of NCI-H295 cells, demonstrating that androgen can induce TGFbeta1 expression and production. TGFbeta1 (10(-7)-10(-6) mol/L) was capable of significantly reducing cell proliferation (P < 0.05) after 24 h of treatment, as assessed by measuring [3H]thymidine incorporation in NCI-H295 cells. The addition of TGFbeta1-neutralizing antibody to cell cultures treated with different DHT concentrations (10(-9) and 10(-10) mol/L) blocked the inhibitory effect of TGF/beta1 on adrenocortical cell proliferation. These findings suggest that TGFbeta1 exerts an inhibitory action on adrenocortical cell proliferation. Therefore, it might be reasonable to suppose that DHT could also influence human adrenocortical cell growth by involving TGFbeta1. PMID- 10690901 TI - No evidence for linkage at candidate type 2 diabetes susceptibility loci on chromosomes 12 and 20 in United Kingdom Caucasians. AB - Several studies have identified evidence for linkage between type 2 diabetes and the regions on chromosomes 12 and 20 containing the maturity-onset diabetes of the young (MODY) genes, hepatocyte nuclear factor-1alpha (HNF-1alpha) and HNF 4alpha. Two studies examining the HNF-1alpha region have demonstrated evidence for linkage at genome-wide levels of significance, whereas four studies examining the HNF-4alpha locus have resulted in evidence for linkage at more suggestive levels of significance. The demonstration of linkage to these regions in additional patient series will strengthen the evidence that susceptibility alleles exist at these loci. We therefore assessed the evidence for linkage to these regions using a large cohort of United Kingdom Caucasian type 2 diabetes affected sibling pairs. A maximum total of 315 affected full sibling pairs were typed for microsatellite markers across the MODY regions and, in a subset of families, for markers spanning the whole of chromosome 20. Evidence for linkage was assessed using a multipoint, mode of inheritance-free method. Linkage analysis did not reveal any significant evidence for excess allele sharing at any of the regions studied. Loci contributing sibling recurrence risks, relative to the general population risk, of 1.75 and 1.25 could be excluded for the HNF 1alpha and HNF-4alpha regions, respectively. We have not confirmed in United Kingdom Caucasians the evidence for linkage previously reported on 12q and 20q. Our results highlight further the problems of replicating previous positive linkage results across different ethnic groups. PMID- 10690902 TI - Increased major histocompatibility complex (MHC) expression in nontoxic goiters is associated with iodide depletion, enhanced ability of the follicular thyroglobulin to increase MHC gene expression, and thyroid autoantibodies. AB - Recent studies suggest that thyroglobulin (TG) accumulated in the follicular lumen of colloid nodular goiters can increase major histocompatibility complex (MHC) class I gene expression in FRTL-5 thyrocytes. Iodide deficiency, also present in these patients, was separately suggested to enhance thyroidal MHC class I and class II gene expression in vivo and in vitro. To test the clinical relevance of these observations, we examined 41 nontoxic goiters surgically removed from patients who had compression problems. Northern analysis revealed that there was a mean 3.9-fold increase in MHC class I expression and a 8.3-fold increase in class II expression by comparison to 9 normal glands. In situ hybridization showed that thyrocytes were the main source of class I and class II transcripts; histological examination revealed that lymphocytic infiltration was minimal to non-existent. The iodine content of the 41 nontoxic goiters was significantly lower than in normal glands, consistent with increased MHC class I and class II. There is also a profound accumulation of TG in the follicles of the nontoxic goiters, and TG purified from the follicles of these glands increased MHC class I gene expression in FRTL-5 thyroid cells significantly more than TG from normal glands per mg protein. Nearly all patients with nontoxic goiter had low, but significantly elevated, levels of antibodies against thyroid peroxidase and/or against TG in their sera compared with those in normal individuals. Moreover, there was a positive correlation between the titer of the serum antibodies against thyroid peroxidase and against TG and MHC class I and class II expression in the thyroid. The data support the possibility that the TG accumulated in the follicular lumen of nontoxic goiters together with relative iodine deficiency contributes to increased MHC expression in thyroid cells in vivo and that increased MHC gene expression contributes to the ability of thyroid antigens to trigger an autoimmune reaction. PMID- 10690903 TI - Vitamin D receptor as a candidate tumor-suppressor gene in severe hyperparathyroidism of uremia. AB - Most chronic renal failure patients with severe refractory hyperparathyroidism harbor at least one monoclonal parathyroid tumor, but the specific acquired genetic defects that confer this clonal selective advantage remain poorly understood. Somatic inactivation of the vitamin D receptor (VDR) gene could contribute to clonal outgrowth, because a parathyroid cell containing this lesion would have an impaired response to the antiproliferative influence of 1,25 dihydroxyvitamin D3. Furthermore, diminished expression of VDR protein has been described in uremia-associated parathyroid tumors. Therefore, to assess VDR gene inactivation's potential pathogenetic role in this disease, we rigorously analyzed the VDR gene in 59 parathyroid tumors surgically resected from uremic patients. First, Southern blotting and/or PCR analyses of 29 tumor samples from 14 genetically informative patients revealed no allelic losses at the VDR locus. Next, direct DNA sequencing of all VDR splice junctions, associated intronic sequences, and virtually the entire VDR-coding region for all 59 tumors revealed no acquired mutations. Last, 37 tumor DNA samples were subjected to comparative genomic hybridization, and no chromosomal losses in the VDR region (12cen-q12) were observed. These observations suggest that inactivating defects within the VDR gene do not commonly contribute to the primary pathogenesis of severe refractory hyperparathyroidism in uremia. PMID- 10690904 TI - Ovarian hyperthecosis in the setting of portal hypertension. AB - Hepatocellular dysfunction and perturbed portal hemodynamics alter steroid metabolism. Men with liver disease have gynecomastia, although women similarly affected rarely show virilization. We report a 10-yr-old girl with portal hypertension and shunting associated with precocious puberty and ovarian hyperandrogenism. This was one of premature twin girls; neither had clitoromegaly or genital ambiguity. In one child, neonatal respiratory problems led to umbilical vein catheterization with subsequent development of portal hypertension. Pubic hair was first noted at age 6 yr, breasts at 7 yr, and severe acne and clitoromegaly at 10 yr. Baseline sex hormones were elevated: androstenedione (A), 413 ng/dL; testosterone (T), 226 ng/dL; and estradiol (E2), 160 pg/mL. Liver transaminases were within the normal range, however, the coagulation profile was mildly abnormal. Cosyntropin adrenal stimulation revealed no steroidogenic defect. Dexamethasone suppression reduced A and T slightly. LH releasing hormone stimulation produced a pubertal rise in LH and FSH. Pelvic sonography showed a large right ovary with numerous follicles. Surgical exploration revealed symmetrically enlarged ovaries with dense capsules. Histology of ovarian wedge resections showed hyperthecosis; immunohistochemistry showed stromal cells expressing steroidogenic enzymes and proteins. One month postoperatively, A and T were unchanged from baseline, whereas E2 decreased to 56 pg/mL. A single dose of depot leuprolide acetate significantly reduced T. Subsequent treatment with oral contraceptives reduced T to 50 ng/dL, and cyclical menses occurred. We conclude that precocious puberty and ovarian hyperthecosis were induced in this young girl by elevated circulating levels of sex hormones, a consequence of portasystemic shunting and impaired hepatic steroid metabolism. PMID- 10690905 TI - Molecular basis off hurthle cell papillary thyroid carcinoma. AB - Among thyroid neoplasms, Hurthle cell tumors (HCTs) have traditionally been a distinct diagnostic category. Hurthle cell adenomas are encapsulated follicular lesions with benign behavior. Hurthle cell carcinomas exhibit unequivocal capsular and/or vascular invasion; they are aggressive tumors with a poor prognosis. Recently, Hurthle cell papillary thyroid carcinomas (PTCs) have been identified on morphological grounds. We hypothesize that a subset of HCTs represent PTC with clinical, histological, and immunohistochemical features based on specific molecular events. ret/PTC gene rearrangements give rise to novel oncogenes that are unique to PTC. We studied a group (n = 50) of HCTs for ret/PTC gene rearrangements. Tumors were examined for papillary differentiation by light microscopic evaluation of nuclear features, by RT-PCR for ret/PTC gene rearrangements, and by immunohistochemistry for ret. Among 24 noninvasive tumors, 13 contained ribonucleic acid for ret/PTC-1, -2, or -3, and 9 of these were immunoreactive for ret. Among 19 Hurthle cell carcinomas, 15 had focal nuclear hypochromasia with grooves and/or inclusions; expressed transcripts of ret/PTC-1, -2, or-3; and exhibited ret positivity. Tumors with ret/PTC gene rearrangements tended to have lymph node metastases rather than hematogenous spread. Our results indicate that a subset of HCTs exhibit features of PTC that are attributable to specific gene rearrangements, resulting in expression of ret/PTC oncogenes. These data support subclassification of HCTs into three groups: Hurthle cell adenomas, Hurthle cell carcinomas, and Hurthle cell PTC. PMID- 10690906 TI - The localization of the functional glucocorticoid receptor alpha in human bone. AB - Glucocorticoids have well-documented effects on the skeleton, although their mechanism of action is still poorly understood. The actions of glucocorticoids on bone cells are mediated, in part, directly via specific receptors. The presence of these receptors has been demonstrated in both rodent and human osteoblastic cells in vitro, but their presence in human bone in vivo has not been reported. In this study, we have used specific affinity purified polyclonal antibodies to the functional glucocorticoid receptor alpha (GRalpha) to investigate its expression in both developing and adult human bone using sections of neonatal rib, calvarial, and vertebral bones, tibial growth plates from adolescents, and iliac crest biopsies from adults who were to undergo liver transplantation. In the tibial growth plates, GRalpha was predominantly expressed in the hypertrophic chondrocytes within the cartilage. In the primary spongiosa, the receptor was highly expressed by osteoblasts at sites of bone modeling. Within the bone marrow, receptors were also detected in mononuclear cells and in endothelial cells of blood vessels. In the neonatal rib and vertebrae, GRalpha was widely distributed at sites of endochondral bone formation in resting, proliferating, mature, and hypertrophic chondrocytes. They were also highly expressed in osteoblasts at sites of bone modeling. At sites of intramembranous ossification in neonatal calvarial bone and rib periosteum, GRa was widely expressed in cells within the fibrous tissue and in osteoblasts at both the bone-forming surface and at modeling sites. In the iliac crests from adults, GRalpha was predominantly expressed in osteocytes. The receptors were not detected in osteoclasts. Our results show for the first time the presence of the functional GRalpha in human bone in situ and suggest that the actions of glucocorticoids on bone may be mediated, in part, directly via the GR at different stages of life. The absence of receptor expression in osteoclasts also suggests that the effects of glucocorticoids on bone resorption may be mediated indirectly. PMID- 10690907 TI - Alpha1 connexin 43 gap junctions are decreased in human adrenocortical tumors. AB - Gap junctional communication disorders have been implicated in the etiology of benign and malignant tumors. Understanding the type, distribution, and frequency of gap junctions in adrenal disorders should provide insight into the role of gap junctions in adrenal carcinogenesis as well as information that may be useful in developing improved diagnosis and treatment of adrenal diseases. Using immunocytochemical techniques, we have characterized and compared alpha1 connexins 43 gap junction protein levels in normal adrenal glands to those in benign and malignant adrenocortical human tumors. In addition, gap junction protein levels were studied in a human adrenal cancer cell line (H295). In both normal and neoplastic adrenal tissues, only alpha1 connexin 43 could be detected, whereas beta1 connexin 32 and beta2 connexin 26 were not found. In the normal adrenal gland, the zona fasciculata was demonstrated to have the highest number of gap junctions per cell (mean +/- SEM, 13.78 +/- 1.93). In contrast, in benign adrenocortical adenomas, the number of gap junctions per cell compared to that detected in normal adrenal glands was significantly reduced (mean +/- SEM, 4.6 +/ 1.17; P < or = 0.05), and the lowest number was found in malignant adrenocortical tumors (1.42 +/- 0.58; P < or = 0.05). Similarly, there were few or no alpha1 connexin 43 gap junctions in the H295 population. There was a progressive decrease in gap junction plaques in adrenocortical cancer cell populations compared to those in normal cell populations. Therefore, analysis of gap junction protein may be helpful for the differential diagnosis of benign and malignant adrenal tumors. The induction of gap junctions in malignant cells may provide a novel therapeutic strategy for adrenal cancer. PMID- 10690908 TI - Estradiol amplifies interleukin-1-induced monocyte chemotactic protein-1 expression by ectopic endometrial cells of women with endometriosis. AB - Endometriosis, one of the most frequently occurring gynecological disorders, is estrogen dependent and is often associated with immunological changes. These include increased macrophage activation and infiltration into the endometriotic implants themselves as well as the peritoneal cavity where the implants often develop. Despite the critical role estrogens play in the development of endometriosis, the biochemical mechanisms of their action remain unclear. In the present study we report that estradiol (E2) enhances endometriotic cell responsiveness to the proinflammatory cytokine interleukin-1beta by up-regulating interleukin-1-induced monocyte chemotactic protein-1 (MCP-1) expression at the level of both protein secretion and messenger ribonucleic acid (mRNA) synthesis, whereas progesterone had no significant effects. According to mRNA half-life experiments, E2 action does not seem to be due to increased MCP-1 mRNA stability but, rather, to a higher level of transcription, as shown by run-on analysis. Interestingly, immunohistochemical analysis of MCP-1 expression in endometriotic tissue showed intense immunostaining in both epithelial glands and stroma regardless of the menstrual cycle phase, which is consistent with the cell culture data and indicates that MCP-1 expression is not subject to cyclic variation. The findings of the present study for the first time provide evidence that E2 up-regulates, although in an indirect way, the expression of a potent chemotactic and activating factor by ectopic endometrial cells, which may occur locally in the inflammatory site and contribute to peritoneal macrophage recruitment and activation, and reveal a new means of E2 action in the pathophysiology of endometriosis. PMID- 10690909 TI - Association of impaired phosphatidylinositol 3-kinase activity in GLUT1 containing vesicles with malinsertion of glucose transporters into the plasma membrane of fibroblasts from a patient with severe insulin resistance and clinical features of Werner syndrome. AB - The purpose of this study was to examine the molecular mechanism responsible for the defective insulin-stimulated glucose transport in cultured fibroblasts from a patient (VH) with clinical features of Werner syndrome and severe insulin resistance. Thus, in cells derived from VH, the subcellular distribution, structure, functional activity, as well as plasma membrane insertion of GLUT1 glucose transporters were analyzed. Furthermore, the insulin signal transduction pathway leading to activation of phosphatidylinositol (PI) 3-kinase as well as components of GLUT1-containing membrane vesicles were characterized. In fibroblasts derived from VH, GLUT1 glucose transporters were overexpressed by 8 fold in plasma membranes (PM) and by 5-fold in high density microsomes, respectively. Exofacial photolabeling revealed that only 14% of the overexpressed PM-GLUT1 transporters were properly inserted into the plasma membrane. The complementary DNA structure of the patient's insulin receptor and the GLUT1 glucose transporter, the intrinsic activity of plasma membrane glucose transporters, the tyrosine phosphorylation, as well as the protein expression of insulin receptor substrate-1/2 and p85 alpha/beta- and p110 alpha/beta-subunits of PI 3-kinase were normal. However, insulin-stimulated association of the p85 subunit of PI 3-kinase was defective in fibroblasts derived from VH compared to those from controls, and this defect was associated with a reduced IRS-1 dependent activation of PI 3-kinase by 50.2% and 63.6% after incubation for 5 and 10 min with 100 nmol/L insulin, respectively. Furthermore, immunodetection of small GTP-binding Rab proteins in subcellular membrane fractions indicated a decreased expression of Rab4 in total cellular homogenates as well as in high density microsomes by 70% and 58%, respectively. After preparation of GLUT1 containing vesicles, Rab4 was not detected to be a component of these vesicles. Analysis of the PI 3-kinase in GLUT1-containing membrane vesicles revealed insulin-dependent targeting of the p85 subunit to the vesicles immunoadsorbed from VH and control fibroblasts. Importantly, the association of the p85 subunit as well as the p85-immunoprecipitable PI 3-kinase activity were markedly reduced in GLUT1-vesicles derived from the patient. In conclusion, impaired PI 3-kinase activity in GLUT1-containing membrane vesicles derived from fibroblasts of VH is associated with a defective docking and/or fusion process of glucose transporters with the plasma membrane and thus might contribute to the molecular defect causing insulin resistance in this patient. PMID- 10690910 TI - Preoperative calcitonin levels are predictive of tumor size and postoperative calcitonin normalization in medullary thyroid carcinoma. Groupe d'Etudes des Tumeurs a Calcitonine (GETC). AB - Medullary thyroid carcinoma (MTC) is a calcitonin (CT)-secreting endocrine tumor. Although plasma CT level is a specific and sensitive marker of MTC, its preoperative usefulness in predicting tumor size and postoperative CT normalization has not been documented. From a nationwide database set up by the French CT Tumor Study Group, 226 MTC patients were selected according to the following criteria: preoperative CT level determination by an immunoradiometric assay (normal value, < 10 pg/mL) within the 6 months prior to surgery, total thyroidectomy and diagnosis of MTC ascertained by histological report including tumor size. Patients were 129 females and 97 males (female/male ratio, 1.3). One hundred and twelve patients (49.6%) had the sporadic variety of the disease, 74 (32.7%) had multiple endocrine neoplasia 2A, three (1.3%) had multiple endocrine neoplasia 2B, and 37 (16.4%) had familial MTC. Median age at diagnosis was 44.8 yr (range, 4.9-80.1 yr). Complete neck dissection was performed in 159 patients (70.4%). Postoperative CT normalization was ascertained by negative response of CT to pentagastrin stimulation (< 10 pg/mL) in 94 patients. Seventy-one patients were considered as not cured because of residual tumor tissue and/or elevated CT levels. Median tumor size was 11.0 mm (range, 0.2-80.0 mm), significantly larger in females (15.0 vs. 8.0 mm, P < 0.05), and in sporadic forms (15.0 vs. 7.0 mm, P < 0.05). Tumor size was significantly correlated (r2 = 0.52, P < 0.01) with preoperative CT levels, the relationship being more straight in familial (r2 = 0.71) than in sporadic (r2 = 0.36) forms. Furthermore, preoperative CT levels under 50 pg/mL appeared to be predictive of postoperative CT normalization (44 of 45 patients). However, higher CT levels did not mean absence of postoperative CT normalization (50 of 120 patients). We conclude that low preoperative CT levels are predictive of tumor size and postoperative CT normalization. PMID- 10690911 TI - Comment on growth hormone therapy and retinal changes mimicking diabetic retinopathy. PMID- 10690912 TI - Emotion and psychopathy: startling new insights. AB - Abnormal affective response in psychopaths is conceptualized within a broad theory of emotion that emphasizes reciprocal appetitive and defensive motivational systems. The startle response is proposed as a specific measure of the directional component of emotional activation. I review the literature that indicates that criminal psychopaths do not show the expected potentiation of the startle reflex that normally occurs during processing of aversive stimuli such as unpleasant photographs or punishment cues. Evidence is presented to demonstrate that this deviant response pattern is specific to individuals who display the classic affective symptoms of psychopathy. The core emotional deviation in psychopathy could be a deficit in fear response, which is defined as a failure of aversive cues to prime normal defensive actions. This emotional deficit may represent an extreme variant of normal temperament. PMID- 10690913 TI - The effect of D-amphetamine, clonidine, and yohimbine on human information processing. AB - Twelve subjects were tested with D-amphetamine, yohimbine, clonidine, and a placebo on a task with two levels of stimulus and two levels of response complexity. The purpose of this study was to test the hypothesis that noradrenergic drugs affect early stimulus processes. D-amphetamine speeded reaction time (RT), clonidine slowed it, and yohimbine had no effect. D amphetamine and yohimbine decreased N1 latency and clonidine increased it. D amphetamine and yohimbine decreased P3 latency and clonidine increased it but, in each case, only when latency estimates were based on single trials, not on averages. D-amphetamine's effect on RT, not P3, as measured by the average, is consistent with previous results. Single trial measures appear more sensitive. Speeding of N1 and single-trial P3 data indicate that noradrenergic drugs affect processing of early (visual) information. D-amphetamine's speeding of single trial P3 estimates was attributed to its noradrenergic actions. Yohimbine's speeding of P3 without changing RT is consistent with neural net (parallel) simulations but not with a serial model. These findings support the assumption that different neurotransmitters modulate specific cognitive processes. PMID- 10690914 TI - Energy expenditure and motor performance relationships in humans learning a motor task. AB - The ability of human subjects to learn minimum energy-demanding variants of biological motion was examined in three adult males trained to walk on hands and feet (creep) on a motor-driven treadmill at constant speed (0.64 m/s) for 16 3 min trials. Two subjects systematically decreased oxygen consumption and heart rate over trials. Following this acquisition phase, subjects completed walking and creeping trials at positive and negative treadmill grades and selected a freely chosen creeping grade that felt "most comfortable." One subject selected a grade that was more efficient than those imposed. Oxygen-consumption curves for walking and creeping converged with increasing positive grade, indicating that increased grade influences the metabolic energy viability of the task (creeping or walking). The acquisition data provide empirical support for the "principle of least effort" and lend support to the concept of a "comfort mode" in the execution of motor tasks. PMID- 10690915 TI - Focusing on the N400: an exploration of selective attention during reading. AB - In a series of two experiments, subjects read sentences wherein words were flanked in the lower visual field by irrelevant words (i.e., flankers). The visual angle between the words in the sentence and the flanker words was manipulated (i.e., 0.57 degrees, 0.97 degrees, 1.37 degrees). Sentence endings were either congruent or incongruent; incongruent endings elicited a large N400 component. Flanker effects were observed for sentence final words on electrophysiological measures during the reading task and on subsequent recognition memory performance for sentence final and new words. For both measures, the flanker effect interacted with the congruency of the sentence ending as well as the visual angle between the sentence final word and its flanker. The largest and earliest flanker effects were observed for congruent endings at the smallest visual angle (0.57 degrees); congruent endings and flankers in intermediate visual angle (0.97 degrees) conditions displayed a similar flanker-related negativity but with a longer onset latency (490 vs. 280 ms). Congruent endings and their flankers in the largest visual angle (1.57 degrees) conditions revealed no flanker effect. PMID- 10690916 TI - Suspense and surprise: on the relationship between expectancies and P3. AB - Few theories of the P3 component have emphasized the distinction between its parietal and frontocentral parts. This study used a new paradigm for testing the predictions that the parietal P3 is evoked by awaited stimuli (suspense) and the frontal P3 by unexpected stimuli (surprise). Subjects had to make simple responses whenever a yellow ring appeared. This signal appeared on the screen within a clock, most frequently when the pointer was at 12 o'clock (every 6 s) but sometimes also at other times. The suspense process should therefore have its minimum shortly after 12 o'clock and then steadily increase until 12 o'clock, and the parietal P3 should accordingly be smallest with stimuli shortly after 12 o'clock, then gradually increase and be largest with 12 o'clock stimuli. Further, the stimuli presented at times other than 12 o'clock should evoke large frontal P3s because they were unexpected. The results confirmed parts of these predictions. A frontocentral and a parietocentral component could indeed be discerned. The frontal P3 was largest with non-12 o'clock stimuli, whereas the parietal P3 was large with all stimuli. The parietal result was not predicted, but these results taken together pose more problems for the usual view, which assumes that the parietal P3 is evoked by unexpected stimuli, than for our assumption that the parietal P3 reflects suspense, and the frontocentral P3 reflects surprise. Generalizing to other paradigms, we assume that different topographies of P3 in different paradigms or in different groups of subjects might be due to different mixtures of these two components. PMID- 10690917 TI - The effect of monocular viewing on heartbeat discrimination. AB - In monocular viewing conditions, an activational imbalance between the cerebral hemispheres was assumed to develop, the direction of which depends on the side of the viewing eye. This assumption was based on the morphological differences between the nasal and the temporal hemiretinas and on physiological data. It was assumed that the hemisphere receiving visual information via the nasal optic fibers, that is, the hemisphere contralateral to the viewing eye, would be the more activated one. Because heartbeat perception is regarded as a predominantly right hemispheric function, it was predicted that during right hemispheric activation created by left monocular viewing heartbeat discrimination performance would be better than during left hemispheric activation created by right monocular viewing. This hypothesis was tested on 30 male right-handed university students who performed a Whitehead-type heartbeat discrimination task while viewing only with the left or with the right eye. Heartbeat perception was more accurate when viewing with the left eye. Additionally, respiratory manipulation during heartbeat discrimination improved performance on this task. PMID- 10690918 TI - Nonconscious associative learning: Pavlovian conditioning of skin conductance responses to masked fear-relevant facial stimuli. AB - We examined the possibility of nonconscious associative learning in a context of skin conductance conditioning, using emotional facial expressions as stimuli. In the first experiment, subjects were conditioned to a backwardly masked angry face that was followed by electric shock, with a masked happy face as the nonreinforced stimulus. In spite of the effectively masked conditioned stimuli, differential conditioned skin conductance responses were observed in a subsequent nonmasked extinction phase. This effect could not be attributed to differential sensitization or pseudo-conditioning. In the second experiment, the differential responding during extinction was replicated with angry but not with happy faces as conditioned stimuli. It was concluded that with fear-relevant facial expressions as the conditioned stimulus, associative learning was possible even in conditions where the subjects remained unaware of the conditioned stimulus and its relationship to the unconditioned stimulus. PMID- 10690919 TI - Developmental dyslexia and attention dysfunction in adults: brain potential indices of information processing. AB - Event-related brain potentials (ERPs) were recorded from a group of 13 men with severe developmental dyslexia and 15 matched normal controls. Auditory and visual stimuli, presented in separate reaction time tasks of graded difficulty, were used to elicit ERPs. No group differences in P300 were seen under relatively undemanding task conditions. However, as task demands increased, visual P300 was reduced in the dyslexic men as compared with the normal readers. An Abbreviated Conners Parent Rating Scale was used to assess retrospectively childhood symptoms of attention-deficit hyperactivity disorder (ADHD). Additional analyses revealed that the dyslexics with a history of many symptoms of ADHD in childhood (high ADHD) accounted for the group differences in P300; the dyslexics with a history of few or no such symptoms (low ADHD) were indistinguishable from the controls at all electrode sites. Furthermore, whereas the low-ADHD dyslexics showed the same hemispheric asymmetry in auditory P300 as did the controls (right > left), auditory P300 was more symmetrically distributed in the high-ADHD dyslexics. The results are interpreted as suggesting that a distinct brain organization may characterize dyslexic men with a history of concomitant deficits in attention. PMID- 10690920 TI - The roles of stimulus novelty and significance in determining the electrodermal orienting response: interactive versus additive approaches. AB - Two competing approaches for the roles of stimulus significance and novelty in determining orienting responses (ORs) were tested and compared. According to the first, ORs to significant stimuli are determined by sets and expectations and are relatively independent of the contrast between the test stimulus and preceding stimuli, whereas responses to nonsignificant test stimuli are products of a Sokolovian match/mismatch process. According to an alternative approach, novelty and significance contribute independently and additively to the OR. Stimulus significance and novelty were independently manipulated in two experiments, and the electrodermal component of the OR was measured while sequences comprising compound pictorial or verbal stimuli were presented. Each sequence included a test stimulus that was either significant or neutral and was preceded by several control stimuli. Novelty was manipulated by varying the contrast between the test and the control stimuli. The results revealing no interaction between stimulus significance and novelty were interpreted as supporting the theory proposed by Cati and Ben-Shakhar (1990). PMID- 10690921 TI - Individual differences in the autonomic origins of heart rate reactivity: the psychometrics of respiratory sinus arrhythmia and preejection period. AB - Heart rate reactivity has been conceptualized, at least implicitly, as a unidimensional construct ranging from low to high, reflecting individual differences in adrenergic reactivity to daily stressors. However, an individual's classification as high in heart rate reactivity ignores possible individual differences in the autonomic origins of this reactivity. Sixty-eight women were exposed to orthostatic and speech stressors to determine the psychometric properties (postural stability, convergent and discriminant validity) of heart rate, preejection period, and respiratory sinus arrhythmia. Results revealed that (a) basal, stress, simple reactivity (stress - baseline), and residualized change indices of heart rate, preejection period, and respiratory sinus arrhythmia were stable across postures and (b) heart rate reactivity was significantly related to preejection period and respiratory sinus arrhythmia reactivity, whereas the latter two measures were unrelated. Reactivity classifications may therefore be significantly improved by attention to concurrent estimates of the activity of both autonomic branches. PMID- 10690922 TI - Statistical mechanic prediction of non-Gomperzian ageing in extremely aged populations. AB - Observations of large populations of ageing organisms have suggested that the exponential increase in the rate of age-related mortality (Gompertz rule) declines in extreme old age, and that the mortality of the extremely long-lived is independent of age. I show that this phenomenon is an expected outcome of ageing being the effect of stochastic failure of highly interconnected networks of elements with constant failure rates, and does not require the postulation of a separate 'long-lived' population of individuals. PMID- 10690923 TI - Satellite cell numbers in senile rat levator ani muscle. AB - Ageing in skeletal muscle results in motor frailty and a reduced capacity for self repair after injury. The contractile characteristics of muscle are determined principally by the myosin heavy chain (MHC) composition of its myofibers. During the restorative process, satellite cells play a central role. The present study compares the levator ani muscle of very old (32 months) and young (4 months) male WI/HicksCar rats in terms of structural integrity, MHC and satellite cell content. Myofiber typing was carried out by indirect immunohistochemistry using a panel of anti-MHC antibodies. Single myofibers for nuclear enumeration were isolated by an enzymatic technique while fiber cross sectional areas and satellite cell frequencies were determined by computerized planimetry and electron microscopy. In both groups of rats, the myofiber population was homogeneously MHC type IIb-reactive. Cross-sectional data reflected a marked degree of atrophy in the muscle of the senile rats (710.05 +/- 63.6 microm2, compared with 1519.98 +/- 79.0 microm2 in young). The myofiber population was reduced by only about 6.7% with ageing and the representation of satellite cells, as a fraction of total sublaminal nuclei, was relatively stable (1.15 versus 1.91% in young; P > 0.05). The results indicate that ageing had a considerable atrophic effect on the levator ani muscle but induced neither MHC isoform transition nor massive depletion of the satellite cell pool. They suggest that the well-documented impairment of the restorative capacity of senile muscle could be due more to alterations in the nature of microenvironmental cues than to quantitative aspects of its cellular capacity to respond. PMID- 10690924 TI - Aging affects the regeneration of the CD8+ T cell compartment in bone marrow transplanted mice. AB - A chimeric mouse model has been used to determine the effect of aging on the differentiation of CD8+ T cells and on the regeneration capacity of the mature peripheral T cell pool after radiation induced depletion. Bone marrow cells from Thy 1.1+ mice were transplanted into lethally irradiated young or aged mice (Thy 1.2+). After 6 weeks, splenic CD8+ T cells were subjected to phenotypic and functional examinations by flow cytometry. Both young and aged mice were able to develop donor derived (Thy 1.1+) CD8+ T cells. Although the absolute number of T cells was reduced in aged recipients, the ratio of CD4+ to CD8+ T cells of donor origin was the same in young Thy 1.1+ control mice as it was in both young and aged chimeric mice, indicating that aging has no effect on the ratio of CD4+ to CD8+ T cells produced by the thymus. However, the percentage of CD8+ cells in the total Thy 1.2+ (host-origin) T cell population was significantly higher in young chimeric mice than in age-matched Thy 1.2+ control mice (P < 0.01), suggesting that a significant over expansion of the Thy 1.2+ CD8+ subset occurred in young mice during regeneration. The Thy 1.1+ CD8+ T cells that developed in young hosts were of a naive phenotype with a majority of cells expressing a low level of CD44. In contrast, the majority of those that developed in the aged host displayed a memory phenotype with a high percentage of cells being CD44hi. In addition, the production of IL-4 and IFN-gamma by Thy 1.1+ CD8+ T cells was affected by the age of the host. A greater fraction of aged Thy 1.1+ CD8+ T cells could be induced to produce either IFN-gamma or IL-4 than young CD8+ T cells. These results suggested that the aged microenvironment has a significant effect on newly developed CD8+ T cells and that the age of the microenvironment also influences the regeneration capacity of CD8+ T cells. PMID- 10690925 TI - Evidence for increased nitric oxide production in the auditory brain stem of the aged dwarf hamster (Phodopus sungorus): an NADPH-diaphorase histochemical study. AB - Age-related changes of the auditory system such as presbyacusis are believed to be due, at least in part, to alterations of central structures. The superior olivary complex (SOC), a group of interrelated brain stem nuclei, projects to a variety of neuronal structures including the cochlea and the inferior colliculus (IC). The soluble gas nitric oxide (NO), believed to function as a neuroactive substance within the SOC and cochlea, is thought to be involved in ageing processes. Since it is unknown whether NO-production is altered in the ageing auditory system, the present study was conducted to investigate whether the number of NO-producing cells in the SOC is changed with increasing age. The histochemical detection of NADPH-diaphorase activity (NADPH-d), a marker for neurons containing NOS, was utilized to investigate the numbers of NO-producing cells in the SOC of adult and senile Djungarian dwarf hamsters (Phodopus sungorus). Our results demonstrate that the number of stained neurons was almost doubled in the SOC of senile hamsters. The most distinct changes were observed in the medial nucleus of the trapezoid body. In contrast, NO-producing preganglionic sympathetic neurons of the spinal intermediolateral nucleus, which was studied for comparison, did not exhibit significant differences between adult and senile animals. It is concluded that the increase of NO-production in the ageing auditory brain stem, as revealed by our data, may be related to hearing impairments with increasing age. PMID- 10690926 TI - A beta and perlecan in rat brain: glial activation, gradual clearance and limited neurotoxicity. AB - A beta1-40 and perlecan (A beta + perlecan) were infused into rat hippocampus for 1 week via osmotic pumps. At the end of the infusion a deposit of A beta immunoreactive material was found surrounding the infusion site. No neurons could be identified within this A beta deposit. The neuron-free area resulting from A beta + perlecan was significantly larger than that found after infusions of A beta40-1 and perlecan (reverse A beta + perlecan), perlecan alone or phosphate buffered saline vehicle. Following infusion of A beta + perlecan, the glial cells segregated in a manner similar to that associated with compacted amyloid plaques in Alzheimer's disease (AD). Activated microglia/macrophages were prevalent within the A beta deposit while the perimeter of the deposit was delimited by reactive astrocytes. Thioflavin S and Congo red staining indicated a beta-pleated sheet conformation of the A beta deposits, implying formation of fibrils. Intact, apparently healthy neurons were found immediately adjacent to the A beta + perlecan deposit. In contrast, reverse A beta peptide did not form congophilic deposits despite the presence of perlecan. Apoptotic profiles visualized with bisbenzamide or TUNEL staining of fragmented DNA were not seen at any of the infusion sites, yet were readily seen in hippocampal sections from animals treated with kainic acid. At 8 weeks, A beta immunoreactivity, Thioflavin S and Congo red staining was reduced, indicating that A beta was being cleared. There also was no evidence of neuron loss by Nissl or TUNEL staining. The zone of apparent necrosis did not expand between 1 and 8 weeks, and in some instances appeared to contract. The consistency of the A beta + perlecan infusion method in producing reliable A beta amyloid deposits permits estimates of the rate at which fibrillar A beta amyloid can be removed from the brain, and may provide a useful model to study this process in vivo. However, the absence of clearly identifiable degenerating/dying neurons at the 1 or 8 week survival times suggests that either fibrillar A beta + perlecan slowly displaced the brain parenchyma during infusion, or neurons were killed very gradually during the process of clearing the A beta. PMID- 10690927 TI - Age-related changes in myelopoietic response to lipopolysaccharide in senescence accelerated (SAM) mice. AB - The effects of in vivo lipopolysaccharide (LPS) administration on myelopoiesis were examined in senescence-accelerated (SAM) mice. Young mice injected with LPS exhibited: (a) increased femoral proliferative pool size; (b) transient reduction in femoral non-proliferative pool size and number of femoral colony forming unit granulocyte macrophages (CFU-GMs); (c) marked increase in splenic CFU-GMs; and (d) transient increase in S-phase of femoral CFU-GMs. The responses of old mice after LPS administration differed from those of young mice in the following points: (a) no recovery of the femoral non-proliferative pool or femoral CFU-GMs, (b) less significant augmentation of the femoral proliferative pool and splenic CFU-GMs, and (c) prolonged reduction in S-phase of femoral CFU-GM. Injection of LPS into mice resulted in a hyperproduction of colony-stimulating activity (CSA) in bone followed by production of colony-inhibitory activity (CIA) in young mice and in contrast, an excessive CIA secretion from bone without an increase in CSA levels in old mice. These imbalances in the regulatory factors derived from non hemopoietic cells in the bones may lead to an inappropriate response of myelopoiesis in aged SAM mice after LPS administration, which may play a key role in infections. PMID- 10690928 TI - The immunology of animal papillomaviruses. AB - Papillomaviruses are species- and tissue-specific double-stranded DNA viruses. These viruses cause epithelial tumours in many animals, including man. Typically, the benign warts undergo spontaneous, immune-mediated regression, most likely effected by T-cells (especially CD4, but also CD8 subsets), whereas humoral immunity can prevent new infections. Some papillomavirus infections fail to regress spontaneously and others progress to malignant epithelial tumours. Additionally, the impact of these lesions is greater in immunosuppressed individuals. Many therapies are ineffective, and there is much interest in the potential for immunological intervention in papillomavirus infections of man and animals. Vaccination can be achieved with 'live' virus, formalin-inactivated virus, synthetic virus-like particles, and DNA vaccination. There has been much recent progress in the development of such vaccines for papillomavirus infections in the rabbit, ox and dog. Success in these animal models suggests that similar approaches may prove useful for prophylactic or therapeutic vaccination against the important human papillomaviruses involved in the development of cutaneous and anogenital warts, laryngeal papillomatosis, and cervical cancer. PMID- 10690929 TI - Differential cytochemical staining characteristics of channel catfish leukocytes identify cell populations in lymphoid organs. AB - This is one of the first characterizations of channel catfish (Ictalurus punctatus) leukocytes by enzyme cytochemistry. Leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, alpha-naphthyl butyrate esterase, beta-glucuronidase, alpha-naphthyl acetate esterase, Sudan Black B and anti-immunoglobulin specific immunohistochemistry. Lymphocytes, monocytes, macrophages, neutrophils, and surface immunoglobulin positive (surface Ig+) cells were present in channel catfish renal hematopoietic tissue and spleen and demonstrated distinctive cytoplasmic foci staining patterns, cytoplasmic blushing or cell membrane staining. Monocytes, macrophages, lymphocytes and surface Ig+ cells were present in the thymus. Thymic and splenic cellular organization appeared very similar to these same mammalian tissues. In the thymus, acid phosphatase positive cells were distributed throughout the parenchyma, while alpha-naphthyl butyrate esterase and beta-glucuronidase positive cells were concentrated in the cortex and the medulla, respectively. Surface immunoglobulin positive cells occurred in the cortex. In the spleen, acid phosphatase positive cells were scattered throughout the parenchyma, while alpha-naphthyl butyrate esterase positive cells were scattered throughout the parenchyma and adjacent to splenic arterioles. Beta glucuronidase and surface immunoglobulin positive cells were restricted to immediately adjacent to splenic arterioles. Sudan Black B positive cells were scattered throughout the parenchyma, while alpha-naphthyl acetate esterase positive cells occurred adjacent to peri-arteriole lymphoid sheaths and appear very similar to mammalian metallophils. PMID- 10690930 TI - Identification of Ostertagia ostertagi specific cells in bovine abomasal lymph nodes. AB - To investigate the contribution of different bovine cell subpopulations in the development of in vitro induced responses by Ostertagia ostertagi third larval antigen extract (L3), bovine abomasal lymph node cell suspensions were depleted of specific cell populations. The depleted cell suspensions were subsequently assayed for their proliferative responses to O. ostertagi L3 antigen extract. Proliferative responses to O. ostertagi L3 antigen extract were restricted to a CD2+ CD4- CD8- cell population and MHC II+ cells different from B-cells were of major importance. Depletion of CD4, CD8, CD4CD8, IgM or CD21 positive cells did not decrease proliferation to L3 antigen extract. Depletion of gammadelta T cells, which also comprise a subpopulation of CD2+ CD4- CD8- cells, reduced proliferation to L3 antigen extract only in one animal. The results suggest that either gammadelta T-cells could be involved in the proliferation or that another as yet unidentified population is important for proliferation. The precise role of these populations during infection with O. ostertagi and the mechanism by which these cells may influence the host immune response are important issues that remain to be elucidated. PMID- 10690931 TI - Characterisation of monoclonal antibodies to ovine interleukin-6 and the development of a sensitive capture ELISA. AB - A purified recombinant ovine (rOv) interleukin-6 (IL-6) was used to generate specific murine monoclonal antibodies (mAbs) and a polyclonal rabbit antisera to this cytokine. From the 31 initial hybridoma cell lines generated, three stable clones were established which secreted mAbs to rOvIL-6, as judged by a direct enzyme-linked immunosorbent assay (ELISA) and Western blotting. Their specificity was further confirmed by demonstrating that none of the mAbs recognised any of the six other irrelevant recombinant ovine cytokines tested by direct ELISA. All three mAbs displayed cross-reactivity with human and African green monkey IL-6 as demonstrated by direct ELISA and Western blotting. In contrast, the polyclonal antibodies only cross-reacted with bovine IL-6 and not with either of the human or monkey homologues. By combining a mAb with the polyclonal antisera a sensitive, IL-6-specific, capture ELISA was developed that had a sensitivity of 150 pg/ml. This detection system was unequivocally validated by demonstrating that native OvIL-6 could be detected in efferent lymph draining from a stimulated popliteal lymph node. In addition, one of the mAbs was shown to allow the detection of OvIL-6 by intracellular cytokine staining and flow cytometry. PMID- 10690932 TI - A standardized gating technique for the generation of flow cytometry data for normal canine and normal feline blood lymphocytes. AB - Flow cytometry is becoming a commonly used technique to characterize a variety of cells. It provides a powerful application to rapidly determine the relative percentages of T-lymphocyte subsets and B-lymphocytes. The effectiveness of its application, however, is dependent on standardization, especially in a clinical setting. Application of flow cytometry to veterinary diagnostics has been limited by the unavailability of reagents and by the unstandardized characterization of normal values using antibodies not commercially available, but typically provided through the generosity of other researchers. This paper presents a standardized gating protocol, and average values and ranges observed for normal canine and feline blood lymphocytes using commercially available antibodies to cell surface markers for CD5, CD3, CD4, CD8, MHC II, and B lymphocytes. The averages for these markers on gated lymphocytes were as follows: Canine CD5 83.3%, Canine CD4 45.0%, Canine CD8 28.8%, Canine MHC II 98.0%, Canine B Cell 12.9%, Canine CD4/CD8 ratio 1.87, Feline T lymphocytes 77.3%, Feline CD4 44.5%, Feline CD8 25.7%, Feline B Cell 24.1%, Feline CD4/CD8 Ratio 1.75. Normal values were also established for a mixed breed group of dogs, and old versus young dogs. This information will provide researchers and clinicians with a standardized protocol for gating, which establishes a basis for comparison between techniques, and a measure of phenotypic percentages for flow cytometry in normal dogs and cats based on this standardization and commercially available antibodies. PMID- 10690933 TI - Cloning, sequencing, and analysis of cDNA encoding bovine granulocyte-colony stimulating factor. AB - Neutrophils play a critical role in defending against bacterial infections. Hematopoietic growth factors are a class of regulatory cytokines that are required for stimulation, proliferation, and differentiation of blood cells. Granulocyte colony stimulating factor (G-CSF) is a cytokine that induces proliferation and maturation of precursor myeloid cells in the bone marrow into fully differentiated neutrophils. G-CSF also modulates the functional activity of mature neutrophils. Treatment with G-CSF significantly enhances neutrophil phagocytic activity and killing of bacteria and fungi. We have isolated and sequenced a cDNA clone encoding bovine G-CSF (bG-CSF) from an endothelial cell cDNA library using primers designed from ovine G-CSF. The full length cDNA is 1460 nucleotides with 585 nucleotides comprising the open reading frame. Sequence analysis shows 95% identity with ovine, 89% with porcine, 85% with human, and 76% with murine G-CSF. The deduced G-CSF protein consists of 174 amino acids with 95% identity to ovine, 86% to porcine, 81% to human, and 71% to murine. The signal peptide of G-CSF is 21 amino acids long which is nine amino acids shorter than that of human and murine G-CSF. RT-PCR analysis shows that neither freshly isolated nor ConA stimulated neutrophils express G-CSF mRNA. Mononuclear cells, however, expressed G-CSF mRNA after 48 h incubation with or without ConA stimulation. PMID- 10690935 TI - Effect of insulin on human aortic endothelial nitric oxide synthase. AB - It has recently been shown that insulin induces vasodilation in human arteries and veins in vivo. This effect of insulin has been shown to be a direct one on the human vein. In view of these observations and the fact that insulin-induced vasodilation is impaired in insulin-resistant states like type 2 diabetes and obesity, we have investigated the hypothesis that insulin may induce the expression of endothelial nitric oxide synthase (e-NOS) in endothelial cells grown from human aortae (HAECs), human lower-limb veins, and human umbilical veins (HUVECs), and microvascular endothelial cells (MVECs) from human adipose tissue. The expression of e-NOS was maximal in HAECs, and therefore, further experiments were performed on these cells. When cells reached 90% confluence, they were induced with different concentrations of insulin (0, 25, 100, and 1,000 microU/mL) for 6 days. The cells were homogenized and e-NOS expression was examined by Western blotting. A dose-dependent induction by insulin of e-NOS in the endothelial cells was clearly demonstrated. There was no detectable level of the inducible NOS isoform (i-NOS), and this effect of insulin was independent of cell proliferation. We conclude that insulin induces a dose-dependent induction of e-NOS in human aortic cells (and possibly arterial/endothelial cells), and this effect may contribute to the overall vasodilatory effect of insulin. PMID- 10690934 TI - Positive reactions to common allergens in 42 atopic dogs in Japan. AB - Clinically important allergens for the diagnosis and treatment of atopic dermatitis vary geographically. In order to identify the most prevalent allergens in atopic dogs in Japan, 42 dogs with a clinical diagnosis of atopy were tested using both in vivo (intradermal skin test (IDST)) and in vitro (antigen-specific IgE assay) allergy tests. Allergens used for IDST included 26 allergen extracts from eight allergen groups: trees, weeds, grasses, house dust mites (HDM), molds, foods, epithelia, and arthropods. Immunodot assay was used to measure antigen specific IgE against 24 allergens from these eight groups and against fish such as cod and sole. In the 42 dogs, the most common positive allergen reaction was to HDM on both IDST (29/42 dogs or 69%) and in vitro testing (23/42 or 54.8%). The second most frequent positive allergen reaction was to Japanese cedar pollen (21/42 or 50.0% for IDST and 7/42 or 16.7% for in vitro testing). In both tests, less than 20% of dogs had positive reactions to molds or foods. Positive reactions to cat epithelia were frequently found on IDST, but rarely found on in vitro testing. Agreement between the two tests was found in 26 instances: HDM (21 dogs), Japanese cedar pollen (five dogs) and wheat (one dog). In this study, the two most common allergens involved in atopic dermatitis in dogs in Japan were HDM and Japanese cedar pollen. PMID- 10690936 TI - Insulin regulation of plasma free fatty acid concentrations is abnormal in healthy subjects with muscle insulin resistance. AB - This study evaluated the ability of insulin to regulate free fatty acid (FFA) concentrations in healthy nondiabetic subjects selected to be either insulin resistant or -sensitive on the basis of insulin-mediated glucose disposal by muscle. Comparisons of steady-state plasma glucose (SSPG), insulin (SSPI), and FFA concentrations were made at the end of 3 infusion periods: (1) under basal insulin conditions (approximately 10 microU/mL), (2) in response to isoproterenol induced stimulation of lipolysis at the same basal insulin concentration, and (3) following inhibition of isoproterenol-induced lipolysis by a 2-fold increase in the insulin concentration. The results showed that steady-state FFA concentrations were significantly higher under basal conditions (360 +/- 73 v 158 +/- 36 microEq/L, P = .02), in response to isoproterenol-induced lipolysis (809 +/- 92 v433 +/- 65 microEq/L, P = .005), and following insulin inhibition of isoproterenol-induced lipolysis (309 +/- 65 v 159 +/- 37 microEq/L, P = .06). These differences were found despite the fact that SSPG concentrations were also higher in insulin-resistant individuals during all 3 infusion periods. These results demonstrate that the ability of insulin to regulate plasma FFA concentrations is impaired in healthy subjects with muscle insulin resistance, indicating that insulin-resistant individuals share defects in the ability of insulin to stimulate muscle glucose disposal and to inhibit adipose tissue lipolysis. PMID- 10690937 TI - Effects of galanin on growth hormone and prolactin secretion in anorexia nervosa. AB - Galanin (GAL) elicits growth hormone (GH) release in normal subjects through interaction with hypothalamic somatostatin. GAL also stimulates GH-releasing hormone (GHRH) secretion in vitro. In rats, GAL is able to stimulate prolactin (PRL) release, but this effect is not clear in humans. We have thus investigated GAL effects on GH and PRL release in patients with anorexia nervosa (AN), known to have altered regulation of the GH-insulin-like growth factor axis and PRL dynamics, and compared the effects of GHRH and GAL on GH and PRL secretion in AN and normal healthy subjects. Eight women with AN (15 to 27 years; body mass index [BMI], 17 to 19.5 kg/m2) were treated with (1) GHRH 50 microg intravenous (IV) injection, (2) porcine GAL 500 microg infusion from -10 to +30 minutes, and (3) 135-minutes saline infusion as a control, respectively. Both peptides induced a significant increase in plasma GH in AN patients (peak level, 27.41 +/- 5.50 microg/L after GAL and 18.97 +/- 2.67 microg/L after GHRH). When data for AN patients and the control group were compared, GH peak levels after GAL were significantly higher in AN patients (27.41 +/- 5.50 v 13.64 +/- 2.32 microg/L), while GH peak levels after GHRH were not different between the 2 groups (18.97 +/ 2.67 v 15.98 +/- 3.88 microg/L). PRL levels significantly increased after both GHRH (peak, 11.70 +/- 2.80 microg/L) and GAL (peak, 18.02 +/- 5.10 microg/L) treatment in AN patients, but not in normal subjects. We conclude that GAL stimulates exaggerated GH release in AN patients as compared with normal controls, suggesting a dual hypothalamic interaction via both an increase in endogenous GHRH and a decrease in somatostatin secretion. Finally, GAL may act as a PRL secretagogue in AN patients. PMID- 10690938 TI - Evaluation of oxidative stress before and after control of glycemia and after vitamin E supplementation in diabetic patients. AB - The present study evaluates the presence of oxidative stress in the uncontrolled diabetic state. Glycemic control reduced the oxidative stress, but total normalization of the parameters of oxidative stress was not achieved, indicating continued oxidant injury despite optimal control of the diabetes. Vitamin E supplementation for 4 weeks in these patients further reduced the oxidative stress, suggesting that vitamin E supplementation might be helpful in reducing free-radical-induced oxidant injury. PMID- 10690939 TI - Inhibitory effect of troglitazone on tumor necrosis factor alpha-induced expression of monocyte chemoattractant protein-1 in human mesangial cells. AB - Insulin resistance is one of the risk factors for the progression of atherosclerosis and glomerulosclerosis. Recently, the new oral insulin sensitizing agent troglitazone has been thought to offer potential in the treatment of diabetes. If adopted for this use, it might be helpful in protecting against the development of atherosclerosis and microvascular complications via its improvement of insulin resistance. However, it has not yet been clarified whether troglitazone acts directly on the vascular cells and inhibits the progression of atherosclerosis, including glomerulosclerosis. Meanwhile, monocyte chemoattractant protein-1 (MCP-1) is known to play an important role in the pathogenesis of atherosclerosis and glomerulosclerosis through the induction of monocyte migration. Therefore, we investigated the effect of troglitazone on the expression of MCP-1 in human mesangial cells (HMCs). HMCs were treated with or without troglitazone (1 or 10 micromol/L) in the presence or absence of tumor necrosis factor alpha (TNF-alpha) at various concentrations (50 or 500 ng/mL), and then MCP-1 secretion from the HMCs was measured. We found that TNF-alpha increased the secretion of MCP-1 by 55-fold versus the control and troglitazone significantly inhibited this TNF-alpha-induced increase in MCP-1 secretion (49.3%). Moreover, Northern blot analysis showed that troglitazone decreased the MCP-1 mRNA level in HMCs. We demonstrated that alpha-tocopherol also inhibited TNF-alpha-induced MCP-1 production in HMCs, although its effects were not as strong as troglitazone. The present study indicates that troglitazone may prevent the progression of atherosclerosis by inhibiting MCP-1 expression in mesangial cells. PMID- 10690941 TI - Metabolism of [1,3-13C]glycerol-1,2,3-tris(methylsuccinate) and glycerol-1,2,3 tris(methyl[2,3-13C]succinate) in rat hepatocytes. AB - Hepatocytes prepared from overnight-fasted rats were incubated for 120 minutes in the presence of 2.5 mmol/L [1,3-13C]glycerol-1,2,3-tris(methylsuccinate) or glycerol-1,2,3-tris(methyl[2,3-13C]succinate). The identification and quantification of 13C-enriched metabolites by a recently developed method for the deconvolution of nuclear magnetic resonance (NMR) spectra with multiplet structures and constraints documented a virtually complete recovery of [1,3 13C]glycerol-1,2,3-tris(methylsuccinate) in 13C-labeled glycerol, lactic acid, and glucose. In hepatocytes exposed to [1,3-13C]glycerol-1,2,3 tris(methylsuccinate), glucose was symmetrically labeled, with the vast majority of hexose molecules being enriched with 13C on both C1 and C3 and/or C6 and C4. The respective abundance of glucose isotopomers labeled either on both C3 and C4 or on only 1 of these 2 C atoms indicated that the triose phosphates generated from [1,3-13C]glycerol represented 44% +/- 1% of the total amount of triose phosphates incorporated into the hexose. In hepatocytes exposed to glycerol-1,2,3 tris(methyl[2,3-13C]succinate), the recovery of [2,3-13C]succinate, [2,3 13C]fumarate, and either double- or single-labeled malate, lactate, alanine, and glucose accounted for about half the initial 13C content of the ester. The majority of the glucose molecules were now labeled in both C, and C2 or C6 and C5, with a preferential labeling of C6-C5 relative to C1-C2, the paired C6/C1 and C5/C2 ratios averaging 1.33 +/-0.04. These findings show that glycerol-1,2,3 tris(methylsuccinate) is efficiently and extensively metabolized in hepatocytes. They reinforce the concept that the asymmetry of glucose 13C-labeling by triose phosphates generated from Krebs cycle intermediates is modulated by the availability of glycerol-derived triose phosphates. Lastly, the present study indicates that the latter triose esters, under the present experimental conditions which do not aim at duplicating the physiological in vivo situation, are largely directly channelled in the gluconeogenic pathway, with only a limited intrahepatic contribution of the "indirect" pathway involving their back-and forth interconversion to and from pyruvate. PMID- 10690940 TI - Lipid and apolipoprotein levels and distribution in patients with hypertriglyceridemia: effect of triglyceride reductions with atorvastatin. AB - Atorvastatin is a new hepatic hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase inhibitor that has been demonstrated to be efficacious in reducing both triglyceride (TG) and cholesterol (CHOL) levels in humans. Twenty-seven (N = 27) patients with primary hypertriglyceridemia (TG > 350 mg/dL) were studied before and after 4 weeks on atorvastatin treatment at a dosage of either 20 (n = 16) or 80 (n = 11) mg/d. The present report examines changes in the plasma levels of several apolipoproteins, including apolipoprotein C-II (apoC-II), apoC-III, and apoE, after atorvastatin. Dose-dependent reductions in both CHOL (20.3% v 43.1%) and TG (26.5% v 45.8%) for the low and high dose, respectively, have been reported in these individuals. In addition to the reductions in apoB commonly associated with the use of HMG-CoA reductase inhibitors, significant reductions in apoE (37% and 49%), apoC-II (28% and 42%), and apoC-III (18% and 30%) were observed with this agent at the 20- and 80-mg/d dosage, respectively. Using fast protein liquid chromatography (FPLC) to fractionate whole plasma according to particle size, the effect of atorvastatin on lipid and apolipoprotein distribution in 20 lipoprotein fractions was also determined. Our results indicate that after 4 weeks on atorvastatin, (1) there was a 2-fold increase in the CHOL content as assessed by the CHOL/apoB ratio for 13 subfractions from very low-density lipoprotein (VLDL) to small low-density lipoprotein (LDL); (2) there was a statistically significant reduction in the percentage of plasma apoB associated with VLDL-sized particles (30.5% v 26.8%); (3) there was a preferential reduction in plasma apoE from non-apoB-containing lipoproteins with treatment; (4) the losses of apoC-II and apoC-III, on the other hand, were comparable for all lipoprotein fractions; and (5) the fraction of plasma TG associated with HDL was increased after treatment. These changes in lipids and apolipoproteins did not depend on the dose of atorvastatin. There was, on the other hand, a dose-dependent reduction in cholesteryl ester transfer protein (CETP) activity, defined as the percentage of 3H-cholesteryl oleate transferred from high-density lipoprotein (HDL) to LDL. CETP activity was reduced by 10.3% and 26.4% with the low and high dose of atorvastatin. Together, these composition data would be consistent with a net reduction in the number of TG-rich lipoproteins that may be explained by (1) a reduction in VLDL synthesis, (2) a preferential removal of VLDL without conversion to LDL, and (3) a preferential accelerated removal of a subpopulation of LDL. PMID- 10690942 TI - Pathogenic factors of glucose intolerance in obese Japanese adolescents with type 2 diabetes. AB - We attempted to identify the pathogenic factors involved in the progression to type 2 diabetes in obese Japanese adolescents. Subjects included 18 nondiabetic obese adolescents, 12 obese adolescents with type 2 diabetes on diet therapy, 10 obese adolescents with type 2 diabetes manifesting ketosis at onset or with a history of treatment with hypoglycemic agents, and 26 non-obese adolescent control subjects. The first-phase insulin response (FPIR), glucose disappearance constant (Kg), glucose effectiveness (Sg), and insulin sensitivity (S(I)) were obtained using an insulin-modified frequently sampled intravenous glucose tolerance test (FSIGT) and a minimal model analysis. The disposition index (DI, by FPIR x S(I)) was determined to assess any endogenous insulin effect. The results showed that Kg was decreased significantly (P = .0006) with the progression to severe diabetes in the obese groups. Although S(I) and Sg did not differ significantly among the 3 obese groups, both parameters were significantly lower in each obese group versus the non-obese controls. As a result of the significant decrease in FPIR (P < .0001), the DI decreased (P = .0006) with the progression to severe diabetes in the obese groups. In conclusion, an early manifestation of type 2 diabetes with occasional ketosis at onset may result from beta-cell dysfunction to glucose stimulation. This finding is demonstrated by the relatively low FPIR to decreased S(I) in obese Japanese adolescents, as well as the low Sg as a synergic role in glucose intolerance. The present findings from a Japanese population for pathogenic factors aside from obesity may help us to gain a better understanding of the progression to adolescent, early-onset, obese type 2 diabetes and its severity. PMID- 10690943 TI - Effect of parathyroid adenoma excision on interleukin-6 (IL-6) and IL-2 receptor levels. AB - 1,25-Dihydroxycholecalciferol [1,25(OH)2D], besides its role in calcium and phosphorus homeostasis, is also an important immunoregulatory molecule. Plasma levels of this hormone may be normal or elevated in patients with primary hyperparathyroidism. 1,25(OH)2D has been reported to inhibit production of the cytokines interleukin-2 (IL-2) and IL-6. In the present study, we examined the effect of parathyroid adenoma excision on serum IL-2 receptor (IL-2R) levels and the release and production of IL-2R and IL-6 by peripheral blood lymphocytes (each measurement was performed twice). Ten patients (5 females and 5 males aged 45 to 78 years) with primary hyperparathyroidism were enrolled in the study. The diagnosis of primary hyperparathyroidism was based on the presence of asymptomatic hypercalcemia, hypophosphatemia, and elevated serum intact PTH levels. Three weeks after removal of the parathyroid adenoma, there was a significant increase in the serum level of IL-2R, as well as the PHA-stimulated peripheral blood lymphocyte production of IL-6 and release of IL-2R. The results indicate that the removal of a parathyroid adenoma in patients with primary hyperparathyroidism causes a significant increase in IL-2R and IL-6 levels. The mechanism by which hyperparathyroidism may affect these cytokines and how they seem related to the levels of vitamin D is discussed. PMID- 10690944 TI - Low-dose epinephrine supports plasma glucose in fasted elderly patients with type 2 diabetes. AB - Recent studies indicate that endogenous epinephrine provides protection against hypoglycemia in fasted elderly patients with type 2 diabetes treated with sulfonylureas. To establish a dose-response relationship and further characterize this hormonal action, 10 subjects with type 2 diabetes aged 67 +/- 1.3 years and receiving glyburide 20 mg daily were studied on 3 separate occasions. Saline placebo, half dose epinephrine ([Epi] 0.375 microg/min), and full dose Epi (0.75 microg/min) were infused during the final 10 hours of a 28-hour fast in a paired, randomized single-blind study to simulate physiologic epinephrine levels. Substrate and hormonal parameters and glucose production (Rd), disposal (Rd), and metabolic clearance rates were determined every 30 minutes. In the placebo study, the mean decline in plasma glucose during the final 10 hours of fasting was -2.7 +/- 0.6 mmol/L, compared with -0.3 +/- 0.3 mmol/L in the half dose Epi study and an actual increase in glucose of 1.0 +/- 0.8 mmol/L in the full dose Epi study (P < .001). There was a similar decline in the glucose Ra in all 3 studies, and the glucose Rd was not significantly different among the 3 study conditions. The baseline-adjusted metabolic clearance rate of glucose was significantly decreased during the epinephrine studies compared with the placebo study (P = .01). The concentration of other counterregulatory hormones did not differ between the studies. We conclude that low physiologic concentrations of epinephrine prevent the progressive decline in plasma glucose observed during fasting in elderly sulfonylurea-treated patients with type 2 diabetes. This finding may be attributable to a relative insulin resistance induced by epinephrine, resulting in a decreased rate of glucose clearance by cells. PMID- 10690945 TI - Genome-wide search for genes related to the fat-free body mass in the Quebec family study. AB - Fat-free mass (FFM) consists mostly of skeletal muscle and bone tissues, and identification of the genes and molecular mechanisms involved in the control of FFM would have implications for the understanding of sarcopenia and potentially osteoporesis associated with aging, as well as the response to starvation, refeeding, anorexia, and any other conditions in which lean body mass is important. A genome-wide search for genes related to body leanness has been completed in the Quebec Family Study (QFS). Microsatellite markers (N = 292) from the 22 autosomal chromosomes were typed. The mean spacing of the markers was 11.9 centimorgans (cM) (range, <0.1 to 41). FFM was calculated from percent body fat, derived from underwater weighing, and body weight and was adjusted by regression for age and sex effects before analysis. A maximum of 336 sib pairs or 609 pairs of extended relatives were analyzed using single-point Haseman-Elston regression (SIBPAL and RELPAL) and multipoint variance component (SEGPATH) linkage analyses. Significant linkages were observed on chromosomes 15q25-q26 for a CA repeat within the insulin-like growth factor 1 receptor (IGF1R) gene (Lod score = 3.56) and at 18q12 with D18S877 (Lod score = 3.53) and D18S535 (Lod score = 3.58), 2 markers located 10 cM apart. A moderately significant linkage was also observed on chromosome 7p15.3 with the marker D7S1808 (Lod score = 2.72). The most obvious candidate genes within the regions identified by these linkages include the IGF1R on 15q and neuropeptide Y (NPY) and growth hormone-releasing hormone (GHRH) receptor on 7p. On 18q, the melanocortin receptor 4 (MC4R) is not likely the candidate gene for the observed linkage. This study represents the first genome wide search for genes that may be involved in the regulation of the lean component of body mass in humans. PMID- 10690946 TI - Stevioside acts directly on pancreatic beta cells to secrete insulin: actions independent of cyclic adenosine monophosphate and adenosine triphosphate sensitive K+-channel activity. AB - The natural sweetener stevioside, which is found in the plant Stevia rebaudiana Bertoni, has been used for many years in the treatment of diabetes among Indians in Paraguay and Brazil. However, the mechanism for the blood glucose-lowering effect remains unknown. To elucidate the impact of stevioside and its aglucon steviol on insulin release from normal mouse islets and the beta-cell line INS-1 were used. Both stevioside and steviol (1 nmol/L to 1 mmol/L) dose-dependently enhanced insulin secretion from incubated mouse islets in the presence of 16.7 mmol/L glucose (P < .05). The insulinotropic effects of stevioside and steviol were critically dependent on the prevailing glucose concentration, ie, stevioside (1 mmol/L) and steviol (1 micromol/L) only potentiated insulin secretion at or above 8.3 mmol/L glucose (P < .05). Interestingly, the insulinotropic effects of both stevioside and steviol were preserved in the absence of extracellular Ca2+. During perifusion of islets, stevioside (1 mmol/L) and steviol (1 micromol/L) had a long-lasting and apparently reversible insulinotropic effect in the presence of 16.7 mmol/L glucose (P < .05). To determine if stevioside and steviol act directly on beta cells, the effects on INS-1 cells were also investigated. Stevioside and steviol both potentiated insulin secretion from INS-1 cells (P < .05). Neither stevioside (1 to 100 micromol/L) nor steviol (10 nmol/L to 10 micromol/L) influenced the plasma membrane K+ adenosine triphosphate ((K+)ATP) sensitive channel activity, nor did they alter cyclic adenosine monophosphate (cAMP) levels in islets. In conclusion, stevioside and steviol stimulate insulin secretion via a direct action on beta cells. The results indicate that the compounds may have a potential role as antihyperglycemic agents in the treatment of type 2 diabetes mellitus. PMID- 10690947 TI - A tale of two homocysteines--and two hemodialysis units. AB - Pharmacologic doses of folic acid are commonly used to reduce the hyperhomocysteinemia of end-stage renal disease (ESRD). Vitamin B12 acts at the same metabolic locus as folic acid, but information is lacking about the specific effects of high doses of this vitamin on homocysteine levels in renal failure. We therefore compared the plasma homocysteine concentrations of maintenance hemodialysis patients in two McGill University-affiliated urban tertiary-care medical centers that differed in the use of vitamin B12 and folic acid therapy. Patients in the first hemodialysis unit are routinely prescribed high-dose folic acid (HI-F, 6 mg/d), whereas those in the second unit receive high-dose vitamin B12 in the form of a monthly 1-mg intravenous injection, along with conventional oral folic acid (HI-B12, 1 mg/d). Predialysis homocysteine was 23.4 +/- 6.8 micromol/L (mean +/- SD) in the HI-F unit and 18.2 +/- 6.1 micromol/L in the HI B12 unit (P < .002). Postdialysis homocysteine was 14.5 +/- 4.1 in the HI-F unit and 10.6 +/- 3.4 micromol/L in the HI-B12 unit (P = .0001). Multiple regression analysis indicated that high-dose parenteral vitamin B12 was associated with a lower homocysteine concentration even after controlling for the potential confounders of sex, serum urea, serum creatinine, urea reduction ratio, and plasma cysteine. Because this was a cross-sectional observational study, we cannot exclude the possibility that unidentified factors, rather than the different vitamin therapies, account for the different homocysteine levels in the two units. Careful prospective studies of the homocysteine-lowering effect of high-dose parenteral vitamin B12 in ESRD should be undertaken. PMID- 10690948 TI - Insulin resistance directly correlates with increased saturated fatty acids in skeletal muscle triglycerides. AB - A close relationship between elevated plasma free fatty acid (FFA) levels and insulin resistance is commonly reported in obese subjects. The aim of the present study was to evaluate the role of intramuscular triglyceride (mTG) and FFA levels in insulin sensitivity in 30 nondiabetic normal-weight or obese subjects (18 with body mass index [BMI] = 21.8 +/- 3.3 kg/m2 and 12 with BMI = 34.6 +/- 2.7 kg/m2) who underwent minor abdominal surgery. Body composition was estimated by isotopic dilution, substrate oxidation by indirect calorimetry, and whole-body glucose uptake by euglycemic-hyperinsulinemic clamp (EHC). Glucose uptake (M) value negatively correlated with the MTG level (R2 = -.56, P < .0001), which was increased in obese patients (11.6 +/- 2.2 v 6.2 +/- 1.4 micromol/g wet weight muscle tissue, P < .0001). The TG fatty acid profile was significantly different in the 2 groups: an increased concentration of saturated fat was present in obese patients (unsaturated to saturated ratio, 1.89 +/- 0.40 v2.19 +/- 0.07, P < .0001). Stepwise linear regression analysis of total mTGs and palmitic and oleic fractions on the M value showed that only TGs and palmitic acid were significantly related to glucose uptake (R2 = .66, P < .0001). Furthermore, among the other anthropometric variables, only the BMI was significantly correlated with MTGs (R2 = .71, P < .0001). In conclusion, not only the MTG concentration but also the FFA pattern seems to affect insulin-mediated glucose uptake. A pivotal role might be played by a high saturated fatty acid content in the TGs. PMID- 10690949 TI - Peroxidation indices and total antioxidant capacity in plasma during hyperhomocysteinemia induced by methionine oral loading. AB - Hyperhomocysteinemia is a risk factor for vascular disease, although its mechanism of action is not fully clear. Different experimental studies have suggested that homocysteine (Hcy) exerts a pro-oxidant effect in the presence of metal ions (Fe and Cu). To test for a similar effect in vivo, we studied plasma markers of lipid and protein oxidation during hyperhomocysteinemia induced by an oral methionine load. Twenty-nine subjects (aged 61 +/- 25 years; 17 women), 25 of whom underwent oral methionine (100 mg/kg) loading, were studied; in every case, we measured total plasma Hcy, malondialdehyde (MDA), conjugated dienes (DIE), and oxidized protein ([PTOX] carbonylic groups) in basal conditions and 4, 6, 8, and 24 hours after methionine loading. Four participants acted as controls. In every case, we also measured total plasma antioxidant capacity (ANTOX) in basal conditions and 8 hours after methionine loading. Eight hours after methionine loading, plasma Hcy increased from 17.6 +/- 11.4 to 54.3 +/- 31.6 nmol/mL, PTOX from 0.33 +/- 0.18 to 0.71 +/- 0.33 nmol/mg protein, DIE from 493 +/- 163 to 590 +/-202 optical density units, and MDA from 1.66 +/- 0.81 to 2.1 +/ 0.93 nmol/mL. There was a significant correlation (Spearman's r) between Hcy and both PTOX (r = .86, P = .01) and MDA (r = .47, P < .05) 8 hours after methionine loading. No significant modifications of the plasma parameters were found during the observation period in controls. ANTOX at 8 hours was significantly (paired ttest) reduced in probands (from 1.74 +/- 0.59 to 1.14 +/- 0.55 mmol/mL, P = .014); no significant difference was observed for plasma ANTOX in controls. Hyperhomocysteinemia due to oral methionine loading induced an increase in plasma oxidation markers. In the absence of hyperhomocysteinemia, no significant modifications were observed. These findings, together with the decrease in ANTOX and the corresponding increase in total plasma Hcy, are consistent with a pro oxidant effect of acute hyperhomocysteinemia in vivo. PMID- 10690950 TI - production rates of testosterone in patients with Cushing's syndrome. AB - Testosterone production rates were determined in 16 patients with Cushing's syndrome (4 men and 12 women) using the stable-isotope dilution technique and mass spectrometry. 1alpha,2alpha-D-Testosterone was infused for 10 hours at a dose of 20 microg/h (men) and 0.4 microg/h (women) and blood samples were obtained at 20-minute intervals during the last 4 hours of the observation period. Estimated production rates in men with Cushing's syndrome were 27, 73, 150, and 180 microg/h (mean, 106 +/- 70 microg/h; healthy men [n = 12], 210 +/- 70 microg/h). In the 12 women with Cushing's syndrome, testosterone production rates were 0.3 to 22.3 microg/h (healthy women [n = 5], 4.3 +/- 1.9 microg/h). There was no difference in testosterone production rates in female patients with central (n = 8) versus adrenal (n = 4) Cushing's syndrome. In summary, testosterone production rates are subnormal or low-normal in male patients with endogenous hypercortisolism, but not in female patients with the same disorder. We conclude that testosterone production in men, but not in women, is predominantly of gonadal origin and hence susceptible to a glucocorticoid-induced suppression of gonadotropin secretion. PMID- 10690951 TI - Changes in phosphatidylcholine fatty acid composition are associated with altered skeletal muscle insulin responsiveness in normal man. AB - The fatty acid composition of skeletal muscle cell membrane phospholipids (PLs) is known to influence insulin responsiveness in man. We have recently shown that the fatty acid composition of phosphatidylcholine (PC), and not phosphatidylethanolamine (PE), from skeletal muscle membranes is of particular importance in this relationship. Efforts to alter the PL fatty acid composition in animal models have demonstrated induction of insulin resistance. However, it has been more difficult to determine if changes in insulin sensitivity are associated with changes in the skeletal muscle membrane fatty acid composition of PL in man. Using nicotinic acid (NA), an agent known to induce insulin resistance in man, 9 normal subjects were studied before and after treatment for 1 month. Skeletal muscle membrane fatty acid composition of PC and PE from biopsies of vastus lateralis was correlated with insulin responsiveness using a 3-step hyperinsulinemic-euglycemic clamp. Treatment with NA was associated with a 25% increase in the half-maximal insulin concentration ([ED50] 52.0 +/- 7.5 to 64.6 +/- 9.0 microU/mL, P < .05), consistent with decreased peripheral insulin sensitivity. Significant changes in the fatty acid composition of PC, but not PE, were also observed after NA administration. An increase in the percentage of 16:0 (21% +/- 0.3% to 21.7% +/- 0.4%, P < .05) and decreases in 18:0 (6.2% +/- 0.5% to 5.1% +/- 0.4%, P = .01), long-chain n-3 fatty acids (1.7% +/- 0.2% to 1.4% +/- 0.1%, P < .01), and total polyunsaturated fatty acids ([PUFAs] 8.7% +/- 0.8% to 8.0% +/- 0.8%, P < .05) are consistent with a decrease in fatty acid length and unsaturation in PC following NA administration. The change in ED50 was significantly correlated with the change in PUFAs (r = -.65, P < .05). These studies suggest that the induction of insulin resistance with NA is associated with changes in the fatty acid composition of PC in man. PMID- 10690952 TI - Regulation of lipolysis and leptin biosynthesis in rodent adipose tissue by growth hormone. AB - The present study examined the effects of growth hormone (GH) on lipolysis and leptin release by cultured adipose tissue from rats and mice incubated for 24 hours in primary culture. A stimulation of leptin release by GH in rat adipose tissue was found in the presence of 25 nmol/L dexamethasone, and this was accompanied by a 28% increase in leptin mRNA content. GH stimulated lipolysis in rat adipose tissue in the presence of 0.1 nmol/L CL 316,243. In contrast, basal lipolysis in mouse adipose tissue was stimulated by GH, but this was not accompanied by an increase in leptin release. However, in the presence of insulin plus triiodothyronine (T3), the stimulation of lipolysis by GH was abolished and GH increased leptin release. These results indicate that GH can stimulate leptin release by both mouse and rat adipose tissue in the absence of a stimulation of lipolysis. In contrast, under conditions in which lipolysis is stimulated by GH, there is no effect on leptin release. PMID- 10690953 TI - Relation of plasma lipids to insulin resistance, nonesterified fatty acid levels, and body fat in men from three ethnic groups: relevance to variation in risk of diabetes and coronary disease. AB - Afro-Caribbean men in the United Kingdom have a favorable lipoprotein profile and are at low risk of coronary heart disease (CHD) compared with Europeans and South Asians, but are at high risk of non-insulin-dependent diabetes mellitus (NIDDM) compared with Europeans. To investigate these differences, a cross-sectional comparison was undertaken for measures of lipoprotein metabolism, body composition, and insulin's glucoregulatory and antilipolytic actions in 92 healthy men (42 to 61 years) of Afro-Caribbean, South Asian, or European origin. Afro-Caribbean men were more insulin-resistant than Europeans (insulin sensitivity [Si], 1.96 v3.01 min(-1) x microU(-1) x mL, P < .01). They nevertheless had a more favorable lipoprotein profile, with lower levels of very low-density lipoprotein (VLDL) cholesterol (0.21 v 0.40 mmol/L, P < .01) and triglycerides (0.34 v 0.74 mmol/L, P < .01), lower serum total triglycerides, higher high-density lipoprotein 2 (HDL2) cholesterol, and larger low-density lipoprotein (LDL) particle size. These differences were not accounted for by differences in nonesterified fatty acid (NEFA) levels, the sensitivity of suppression of NEFA levels to insulin, or body composition. South Asians were also more insulin-resistant than Europeans but had a less favorable lipoprotein profile. Afro-Caribbean men in the United Kingdom are as insulin-resistant as South Asian men but less susceptible to the lipid disturbances that characteristically accompany insulin resistance. This favorable lipid pattern may relate to more effective VLDL metabolism rather than a reduced supply of NEFA as substrate for triglyceride synthesis. PMID- 10690954 TI - Age-related differences in the secretion of calcitonin in male rats. AB - The mechanism of hypercalcitoninemia associated with aging was investigated in male rats. To mimic some of the hormonal changes with aging, orchidectomized (Orch) and hyperprolactinemic rats were used to mimic the physiological status of aging. Orch and haloperidol-induced hyperprolactinemic rats aged 3, 8, and 17 months were infused with CaCl2 and then bled from a jugular catheter following the CaCl2 challenge. Rat thyroid gland was incubated with Locke's medium at 37 degrees C for 30 minutes. Compared with 8- and 3-month-old rats, 17-month-old rats exhibited the lowest levels of plasma testosterone and the highest levels of plasma prolactin (PRL) and calcitonin (CT). The release of CT in the thyroid glands in vitro was highest in 17-month-old rats. Orchidectomy decreased rat plasma CT and thyroid CT release in vitro. Hyperprolactinemic rats had higher levels of plasma PRL and CT compared with control animals. The release of thyroid CT in vitro was greater in hyperprolactinemic rats. These results suggest that the hypersecretion of CT in 17-month-old rats may be due in part to hyperprolactinemia. PMID- 10690955 TI - Leptin during and after preeclamptic or normal pregnancy: its relation to serum insulin and insulin sensitivity. AB - Hyperleptinemia may be part of the insulin resistance syndrome. We studied serum leptin in preeclampsia, which is an insulin-resistant state, and sought associations between leptin and insulin or insulin sensitivity during and after pregnancy. Twenty-two proteinuric preeclamptic women and 16 normotensive controls were studied during the third trimester. Leptin was higher in preeclampsia (mean +/- SE, 34.6 +/- 3.9 v 20.0 +/- 3.3 microg/L, P = .002) and correlated directly with the level of proteinuria (r = .47, P = .03) and normal pregnancy (r = .52, P = .04), whereas insulin sensitivity as assessed by an intravenous glucose tolerance test showed no relationship to leptin. Leptin was 19.0 +/- 3.6 microg/L in 14 preeclamptic women and 10.1 +/- 2.0 microg/L (P = .11) in 11 controls 3 months after delivery. Leptin correlated directly with insulin both in preeclamptic puerperal women (r = .63, P = .02) and in controls (r = .81, P = .003). Leptin and insulin sensitivity correlated only in preeclamptic puerperal women (r = -.59, P = .02). In conclusion, (1) serum leptin is elevated in preeclampsia, (2) insulin is an important determinant of serum leptin in preeclamptic and normotensive women both during pregnancy and in the puerperium, and (3) hyperleptinemia may be part of the insulin resistance syndrome also in women with prior preeclampsia. PMID- 10690956 TI - Endothelin-1 (ET-1)-potentiated insulin secretion: involvement of protein kinase C and the ET(A) receptor subtype. AB - Endothelin-1 (ET-1), a potent vasoconstrictor peptide of endothelial origin, is capable of influencing hormone secretion from endocrine tissues, eg, pancreatic islet cells. We have shown a direct stimulatory effect of ET-1 on insulin secretion from isolated mouse islets of Langerhans. However, it is unknown as to whether the peptide acts through specific receptors on the islet cells and which mechanisms are involved in this insulinotropic action. We have therefore used the specific ET(A) receptor antagonist BQ123, the ET(B) receptor agonist BQ3020, and classic alpha- and beta-adrenergic and cholinergic antagonists. ET-1 (100 nmol/L) stimulated insulin secretion from islets incubated at 8.3, 11.1, 16.7, and 25 mmol/L glucose (P < .05). At 3.3 mmol/L glucose, no alteration in insulin secretion was found. The cholinergic receptor antagonist atropine (5 micromol/L) or the adrenergic receptor antagonists propranolol (5 micromol/L) or phentolamine (5 micromol/L) did not affect ET-1 (100 nmol/L)-stimulated insulin secretion. BQ123 (10 pmol/L to 10 nmol/L) and BQ3020 (1 nmol/L to 1 micromol/L) had no effect on glucose (16.7 mmol/L)-stimulated insulin secretion, but BQ123 counteracted the stimulatory effect of ET-1 (100 nmol/L) at concentrations of 1 nmol/L to 10 micromol/L (P < .01). We also studied the relative role of protein kinase C (PKC) and a Wortmannin-sensitive pathway for ET-1-induced insulin secretion using 12-O-tetradecanoyl phorbol-13-acetate (TPA), Calphostin C, and Wortmannin, respectively. At 5.6 mmol/L glucose, ET-1 (100 nmol/L) had no effect per se, whereas in the presence of 1 micromol/L TPA, which acutely stimulates PKC, the peptide did potentiate insulin secretion (P < .05). Furthermore, the insulinotropic effect of ET-1 at 16.7 mmol/L glucose was counteracted by the PKC inhibitor Calphostin C (P < .05) and by downregulation of PKC by 24 hours of exposure of islets to TPA (0.5 micromol/L, P < .05). Wortmannin (1 micromol/L) did not alter ET-1-potentiated insulin secretion. In conclusion, our results suggest that ET-1 acts through specific ET-1 receptors, most likely the ETA subtype. Furthermore, PKC plays an essential role in the insulinotropic action of ET-1 in mouse islets. PMID- 10690957 TI - Selective consumption of thyroxine-binding globulin during cardiac bypass surgery. AB - A study of serum thyroid hormone binding proteins and thyroid hormone concentrations during and after coronary artery bypass graft (CABG) surgery shows a marked difference in the response of thyroxine binding globulin (TBG) and transthyretin (TTR). The effects of CABG on TBG and TTR were compared in 32 patients during the day of surgery. In a few of these patients, additional determinations were performed to 5 days. When corrected for dilution, TTR concentrations decline gradually after surgery, with no significant decrease over the first 24 hours. In contrast, a rapid decrease of TBG to a mean level of 60% of the preoperative control at 12 hours after the start of surgery appears to account for the concomitant decrease of serum T4. The rate at which the TBG concentration decreased far exceeds the reported fractional clearance of TBG and therefore implies accelerated consumption rather than inhibition of production. TBG is a member of the serine protease inhibitor (SERPIN) superfamily. We propose that its rapid consumption is due to protease cleavage at inflammatory sites. This may explain the previously observed accumulation of thyroxine iodine at such sites. PMID- 10690958 TI - Changes in adenylyl cyclase isoforms as a mechanism for thyroid hormone modulation of cardiac beta-adrenergic receptor responsiveness. AB - Although thyroid hormones are known to modulate cardiac beta-adrenergic receptor expression, the physiologic implications of these changes in the cardiac manifestations of altered thyroid hormone metabolism have been disputed. This study examined whether thyroid hormone modulates signaling via the cyclic adenosine monophosphate (cAMP) pathway by regulating cardiac adenylyl cyclase (AC) isoform expression. Northern blot analyses and AC enzyme assays were performed on preparations from hypothyroid, euthyroid, and hyperthyroid rat ventricles. Steady-state levels of cardiac AC mRNA types V and VI in hypothyroid ventricles were 173% +/- 8% and 149% +/- 12%, respectively, of the values in euthyroid ventricles (P < .01). This increase in AC mRNA isoforms was accompanied by a 1.5-fold increase (P < .05) in the activation of catalytic AC by forskolin and Mn. In contrast, the relative abundance of transcripts for types V and VI AC was similar in hyperthyroid and euthyroid ventricles, but catalytic AC activation by forskolin and Mn was significantly reduced by 35% in membranes obtained from hyperthyroid ventricles. AC activation through beta-adrenergic receptor stimulation by isoproterenol was not altered by thyroid hormone status. Thus, the effect of thyroid hormone to repress AC catalytic activity would be anticipated to offset the increase in beta-adrenergic receptor expression in hyperthyroidism. These studies identify cardiac AC enzymes as important targets for thyroid hormone-dependent regulation of signaling via the cAMP pathway, and support the finding that cardiac adrenergic responsiveness is unaltered in thyroid disease states. PMID- 10690959 TI - Molecular scanning analysis of hepatocyte nuclear factor 1alpha (TCF1) gene in typical familial type 2 diabetes in African Americans. AB - Type 2 diabetes mellitus (T2DM) is strongly inherited, but the major genes for this disease have been elusive. In contrast, early-onset, autosomal-dominant diabetes results from at least 5 loci, of which hepatocyte nuclear factor 1a (HNF1alpha or TCF1) is the most common cause. Mutations in HNF1alpha also cause later-onset diabetes in some Caucasian populations, but the role of these mutations has not been tested in African American populations. We used a variety of screening methods, including both single-strand conformation polymorphism (SSCP) analysis and dideoxy fingerprint analysis, to search for mutations in 51 African American subjects with onset of diabetes before age 50 years. Potential mutations were confirmed by direct sequencing. We identified 21 different variants, of which 11 were unique to African Americans. Four mutations either altered the amino acid sequence (Gly52Ala and Gly574Ser) or were close to a splice site (intron 1 and intron 10). A 5-nucleotide insertion in intron 1 was present in both diabetic members of a small family, but Gly52Ala, Gly574Ser, and the intron 10 mutation did not segregate with diabetes. Gly574Ser was present in 2 large families and 5% of controls, all of which appeared to share the same common HNF1alpha haplotype. Surprisingly, radioactive SSCP analysis under 2 room temperature conditions performed as well as methods using fluorescent labeling that were expected to be more sensitive. We conclude that in African American individuals under age 50, variation in the HNF1a gene is common but unlikely to be a significant cause of T2DM. PMID- 10690961 TI - Roll over Pick and tell Alzheimer the news! PMID- 10690960 TI - Brain atrophy as a surrogate marker in MS: faster, simpler, better? PMID- 10690962 TI - A stimulating view of human visual cortex. PMID- 10690963 TI - Neurology in the next two decades: report of the Workforce Task Force of the American Academy of Neurology. PMID- 10690964 TI - HMG-CoA reductase inhibitors (statins): a promising approach to stroke prevention. AB - Statins represent a promising class of agents to prevent stroke. In randomized trials of middle-aged patients with coronary artery disease, statins reduce the incidence of stroke. The reduction in stroke may not be solely related to cholesterol or low-density lipoprotein reduction but may involve nonsterol mechanisms effects on endothelial cells, macrophages, platelets, and smooth muscle cells. Statins also reduce the size of cerebral infarction in a murine stroke model, suggesting a neuroprotective effect. The best current evidence for stroke prevention is with pravastatin and simvastatin. Pravastatin reduces the risk of stroke in patients with coronary artery disease and average cholesterol levels; simvastatin reduces the risk of the combined endpoint of stroke and transient ischemic attack in hypercholesterolemic patients with coronary artery disease. Future studies of statins are needed in stroke populations, particularly the elderly. PMID- 10690965 TI - Brain embolism is a dynamic process with variable characteristics. PMID- 10690966 TI - Intrarater and interrater reliability of the MS functional composite outcome measure. AB - OBJECTIVE: To assess practice effects, and intrarater and interrater reliability of the MS functional composite (MSFC) outcome measure. BACKGROUND: To address the poor reliability and insensitivity to change of available MS clinical rating scales, the National MS Society's Clinical Outcomes Assessment Task Force developed the MSFC, a multidimensional quantitative clinical outcome measure that includes tests of leg function/ambulation (Timed 25-Foot Walk), arm function (Nine-Hole Peg Test), and cognitive function (Paced Auditory Serial Addition Test). METHODS: Ten patients with secondary progressive MS underwent six testing sessions over a 2-week period. The MSFC was administered by the same examining technician in the first five sessions and by the other technician in the sixth. Patients were reassessed by both technicians after 6 months (sessions 7 and 8). The MSFC score was calculated as the mean of the Z scores of the three components. A pooled dataset derived from secondary progressive MS patients in the placebo arms of previous clinical trials and natural history studies served as the reference population to standardize scores. RESULTS: Practice effects were evident initially but stabilized by the fourth administration. The intraclass correlation coefficient (ICC) was 0.97 for the MSFC for session 4 versus session 5 (intrarater reliability). The ICC was 0.95 for session 5 versus session 6 (interrater reliability), and was 0.96 for session 7 versus session 8 when patients were reassessed 6 months later. CONCLUSIONS: The MS functional composite (MSFC) outcome measure had excellent intrarater and interrater reliability when standardized procedures were used to train examining technicians and to assess patients. Prebaseline testing sessions should be included in clinical trials employing the MSFC to compensate for practice effects. PMID- 10690967 TI - Progressive cerebral atrophy in MS: a serial study using registered, volumetric MRI. AB - OBJECTIVE: To assess the potential of registered volumetric MRI in measuring rates of atrophy in MS. BACKGROUND: Pathologic and imaging studies suggest that the development of permanent neurologic impairment in MS is associated with progressive brain and spinal cord atrophy. Atrophy has been suggested as a potential marker of disease progression. Conventional atrophy measurements requiring manual outlining are time-consuming and subject to reproducibility problems. Registration of serial MRI may offer a useful alternative in that cerebral losses may be measured directly from automated subtraction of brain volumes. METHODS: Twenty-six patients with MS and 26 age- and gender-matched controls had two volumetric brain MR studies 1 year apart. Baseline brain and ventricular volumes were measured using semiautomated techniques, and follow-up scans were registered to baseline. Rates of cerebral atrophy were calculated directly from the registered scans. RESULTS: Baseline brain volumes in the MS group were smaller (mean difference 78 mL [95% CI 13 to 143; p = 0.02]) and ventricular volumes greater (mean difference 12 mL [95% CI 6 to 18; p < 0.001]) than controls. The rate of cerebral atrophy in the MS group (0.8% per year) was over twice that of controls (0.3%), and the rate of ventricular enlargement was five times greater than the controls (1.6 versus 0.3 mL/year). CONCLUSION: Progressive cerebral atrophy is an important feature of MS. Registration-based measurements are sensitive and reproducible, allowing progressive atrophy to be detected within 1 year and may have potential as a marker of progression in monitoring therapeutic trials. PMID- 10690968 TI - Glatiramer acetate (Copaxone) treatment in relapsing-remitting MS: quantitative MR assessment. AB - OBJECTIVE: To evaluate the efficacy of glatiramer acetate (GA, Copaxone; Teva Pharmaceutical Industries, Ltd., Petah Tiqva, Israel) by MRI-based measures in patients with relapsing-remitting (RR) MS. METHODS: Twenty-seven patients with clinically definite RR-MS were treated with either 20 mg of GA by daily subcutaneous self-injection (n = 14) or placebo (n = 13) for approximately 24 months. Axial dual-echo fast-spin-echo T2-weighted images and T1-weighted images before and after gadolinium (Gd) were acquired at 1.5 tesla and transferred into an image processing computer system. The main outcome measures were the number of Gd-enhanced T1 and T2 lesions and their volume as well as brain parenchyma volume. RESULTS: The values of age, disease duration, Expanded Disability Status Scale (EDSS) score, the number of T1- and T2-weighted lesions, and their volume were similar between GA- and placebo-receiving groups at the entry of this study. There was a decrease in the number of T1-enhanced lesions (p = 0.03) and a significant percent annual decrease of their volume in GA recipients compared with those of placebo recipients. There were no significant differences between changes in the two groups in the number of T2 lesions and their volume. The loss of brain tissue was significantly smaller in the GA group compared with that of the placebo group. CONCLUSIONS: These results show that GA treatment may decrease both lesion inflammation and the rate of brain atrophy in RR-MS. PMID- 10690969 TI - Mitral annulus calcareous brain emboli. PMID- 10690970 TI - Familial frontotemporal dementia with ubiquitin-positive, tau-negative inclusions. AB - OBJECTIVE: To describe the clinical features, neuropathology, and genetic studies in a family with autosomal dominant frontotemporal dementia (FTD). BACKGROUND: Clinical Pick's disease, or FTD with parkinsonism, has been described in several families linked to chromosome 17 (FTDP-17). Most of these have shown tau protein mutations. The clinical and pathologic variations in these families resemble the spectrum of sporadic FTD or "Pick complex." METHODS: Clinical and behavioral analysis of the affected members with extensive histochemical and neuropathologic description of three cases, genetic analysis of three clinically affected members and seven at risk members to assess linkage to chromosome 17, and sequencing of the tau gene in two patients were performed. RESULTS: The clinical pattern shows a highly stereotypic disinhibition dementia with late extrapyramidal features, progressive mutism, and terminal dysphagia in three generations of affected individuals. Neuropathology showed frontotemporal atrophy, and microscopically tau- and synuclein-negative and ubiquitin-positive neuronal inclusions, in the background of superficial cortical spongiosis, neuronal loss, and gliosis. Tau expression was restricted to oligodendroglia. All exons and surrounding introns of the tau gene were sequenced, and no mutation or disease-related polymorphisms were detected in either of two affected pedigree members. CONCLUSION: This family with autosomal dominant frontotemporal dementia (FTD) shows no tau expression in neurons. The ubiquitin-positive, tau-negative inclusions have been described before in FTD with and without motor neuron disease, but not in a familial form. The clinical and some pathologic features are similar to those of several of the families included in descriptions of FTD with parkinsonism linked to chromosome 17, but the linkage to tau has been excluded. The defect in this family, however, could be functionally related to tau mutations. PMID- 10690971 TI - Memory impairment on free and cued selective reminding predicts dementia. AB - OBJECTIVE: To estimate the relative rates of dementia in initially nondemented subjects with and without memory impairment defined by baseline free recall from the Free and Cued Selective Reminding (FCSR) test. BACKGROUND: Our approach to identifying persons at high risk for future dementia is to show the presence of memory impairment not caused by other cognitive deficits by using a memory test that controls attention and cognitive processing. When the conditions of testing are not adequately controlled, prediction is reduced because age-associated memory deficits due to other cognitive deficits are confused with dementia associated memory deficits. METHODS: Longitudinal evaluation of 264 initially nondemented, elderly community volunteers from the Einstein Aging Study with clinical and psychometric examinations every 12 to 18 months for up to 10 years. MAIN OUTCOME MEASURES: Dementia was defined by an algorithmic definition that required a Blessed Information Memory and Concentration score >8 and clinical evidence of functional decline. RESULTS: Thirty-two incident cases of dementia developed during follow-up. Survival analyses indicated that subjects with impaired free recall at baseline had dementia develop (relative risk = 75.2, 95% CI = 9.9 to 567) over 5 years of follow-up at dramatically higher rates than subjects with intact free recall after adjusting for age, gender, and education. CONCLUSION: Poor performance on free recall from FCSR predicts future dementia. These findings support the existence of a preclinical phase of dementia characterized by memory impairment, which is present for at least 5 years before diagnosis. PMID- 10690972 TI - Endogenous estrogen levels and Alzheimer's disease among postmenopausal women. AB - BACKGROUND: Although several studies have suggested that hormone replacement therapy lowers the risk of AD among postmenopausal women, few studies have evaluated the relationship of endogenous estrogen levels and AD. The current study investigated whether serum estrone and estradiol levels were related to the presence of AD among postmenopausal women not currently taking hormone replacement therapy. METHODS: Using a case-control design, we examined an ethnically diverse sample of postmenopausal women who met National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria for AD (n = 50) and nondemented controls (n = 93). All women were participants in a study of aging and dementia and were seen consecutively between August 1997 and October 1998. RESULTS: Patients with AD had lower estradiol (F[1,141] = 8.3, p = 0.005) levels than did normal controls. Patients also had lower estrone levels; however, this comparison did not quite meet significance criteria (F[1,141] = 3.6, p = 0.06). Compared to estradiol levels >20 pg/mL, women with AD were four to six times more likely to have levels <20 pg/mL after adjusting for age, years of education, presence of an APOE-epsilon4 allele, ethnicity, and body mass index. There were no significant differences in frequency of AD among women within different quartiles of estrone after adjusting for potential confounds. CONCLUSIONS: The results of this preliminary case-control study suggest that estradiol levels may decline significantly in women in whom AD develops. PMID- 10690973 TI - White matter volumes and periventricular white matter hyperintensities in aging and dementia. AB - OBJECTIVE: To determine the relationship between MRI periventricular white matter hyperintensities, cerebral white matter volumes, neuropathologic findings, and cognitive status in aged individuals. BACKGROUND: The significance of periventricular white matter hyperintensities seen on MR images in aged individuals remains controversial. The Nun Study is a longitudinal cohort aging study in which all 678 initially enrolled participants agreed to autopsy neuropathologic examination. METHODS: We used MRI to measure white matter volumes of the cerebral hemispheres in 52 formaldehyde-fixed brains for correlation with white matter and neocortical pathology, postmortem MRI observations, and cognitive measures. RESULTS: Reduced white matter volume is associated with dementia, but periventricular white matter hyperintensities were not related to white matter volume, stroke, or dementia. CONCLUSIONS: Our results do not support the hypothesis that periventricular hyperintensities seen on MR images have deleterious consequences in these aged individuals. PMID- 10690974 TI - Pure hippocampal sclerosis: a rare cause of dementia mimicking Alzheimer's disease. AB - OBJECTIVES: To identify patients with pure hippocampal sclerosis (HS) as a cause of dementia, to determine whether they have had histories of hypotension or hypoxia, and to compare the clinical features of patients with pure HS with a control group of AD patients without HS. METHODS: In a retrospective study, the authors reviewed all 1771 cases received in their dementia brain bank from 1978 through 1996 to identify those patients with pure HS, defined as severe degeneration and gliosis of the CA1 sector and subiculum of the hippocampal formation in the absence of other significant dementing disease such as Alzheimer's changes. The control group included all patients received during the same period with severe AD without HS, infarcts, or other dementing disease. RESULTS: Seven pure HS cases (0.4%) were identified. None had any episodes of syncope, hypotension, or hypoxia reported in association with dementia onset. Six had memory loss as the primary presenting symptom, and all became progressively demented. Forty-five AD patients without HS were identified for the control group. There were no clear clinical differences between the two groups with regard to sex, age at onset, risk factors for vascular disease, symptoms of cerebrovascular disease, treatment with tranquilizing medications, treatment for depression, or nursing home placement. There was a tendency for heart disease to be more prevalent and the duration of illness to be shorter in the patients with pure HS. CONCLUSIONS: Pure hippocampal sclerosis (HS) occurred in only 0.4% of our dementia patients. Clinically, the seven patients with pure HS were similar to our AD control group. Further research is needed to determine the causes of HS and why HS appears to mimic AD. PMID- 10690975 TI - Mapping of functional organization in human visual cortex: electrical cortical stimulation. AB - OBJECTIVES: To investigate the pattern of functional organization in the human visual cortex through electrical cortical stimulation. METHODS: Electrical cortical stimulation was applied to the occipital cortex and adjacent cortices using subdural grid electrodes in 23 epilepsy patients. Diverse visual responses were recorded. These responses were divided into different categories according to the specific response modalities, such as form, color, and motion. Form visual responses were further subdivided into simple, intermediate, and complex responses. The cortical localization of subdural electrodes was identified using MRI-CT coregistration. The cortical distribution of different visual responses was projected into three-dimensional surface renderings of the brain. The distribution and frequency of subdural electrodes showing different visual responses were quantified by calculating the percentage of the number of electrodes showing one specific type of visual response at the corresponding anatomic region to the total number of electrodes in all brain regions that produced the same response. RESULTS: Simple form responses were obtained mostly at the occipital pole and the inferior occipital gyrus (47.4%) and the striate cortex (42.4%). Intermediate form responses occurred mainly on the peristriate cortex (52.5%) and the lateral occipital (28.0%) and fusiform gyri (19.5%). Complex forms were produced by stimulation of the basal temporo-occipital region (57.6%) and the lateral temporal or lateral temporo-occipital junctional region (42.4%). Color responses occurred on the basal occipital area, mostly at the fusiform (40.0%) and lingual gyri (36.0%). Moving sensations were evoked by stimulation of the basal temporo-occipital (28.4%) and the mesial parieto occipital or temporo-parieto-occipital junctional regions (23.9%). CONCLUSIONS: Different modalities of vision, such as form, color, and moving sensation, appeared to be distributed and organized in different areas of the human visual cortex. PMID- 10690976 TI - Palinopsia and polyopia in the absence of drugs or cerebral disease. AB - OBJECTIVE: To report the occurrence of palinopsia and polyopia in patients who neither used drugs nor had diseases of the cerebral hemispheres, a group in which these visual symptoms have not been reported. METHOD: The patient records in the database of an academic neuro-ophthalmology unit were reviewed. RESULTS: Seventeen patients were identified in the database with the diagnosis of palinopsia or polyopia, of whom eight had diseases of the cerebral hemispheres, leaving nine patients for analysis. No patients with a history of drug toxicity were identified. In one patient the symptoms presented during an initial episode of demyelinative optic neuritis in the absence of clinical or laboratory evidence of cerebral lesions. In another patient they developed immediately after laser treatment of diabetic macular edema. A third patient developed the symptoms in association with visual loss from Leber's hereditary optic neuropathy. The other six patients were healthy individuals. CONCLUSION: Palinopsia and related visual symptoms can occur in otherwise healthy individuals and in patients with disease apparently confined to the eye or the optic nerve. PMID- 10690977 TI - Dissociation of smooth pursuit and vestibulo-ocular reflex cancellation in SCA-6. AB - OBJECTIVE: To study gaze in SCA-6 patients during pursuit and passive whole-body rotation. BACKGROUND: Smooth pursuit and vestibularly induced eye movements interact to maintain the accuracy of eye movements in space (i.e., gaze). Previous studies have implicated the cerebellum, particularly the floccular lobe and dorsal vermis, in the control of gaze velocity during pursuit and vestibulo ocular reflex (VOR) cancellation. SCA-6 has recently been identified genetically and characterized as pure cerebellar ataxia that affects the cerebellar cortex selectively. METHODS: Using infrared oculography, eye movements of five SCA-6 patients and five age-matched normal control subjects were recorded during sinusoidal pursuit and passive whole-body rotation in the horizontal plane (amplitude, +/- 10 deg; frequency, 0.2 Hz). Eye and gaze gain (eye and gaze velocity/stimulus velocity) were calculated after deleting saccades. RESULTS: Eye gain of all SCA-6 patients during pursuit was significantly lower than those of the control subjects (mean +/- SD, 0.26+/-0.06 versus 0.91+/-0.07). In contrast, eye gain of the patients was not significantly different from that of the control subjects either during VOR cancellation, when the subjects tracked a target that moved with the same amplitude and phase, like a chair (0.21+/-0.05 versus 0.12+/ 0.07), or during visually enhanced VOR (x1), when the target remained stationary in space (0.85+/-0.06 versus 0.95+/-0.05). Moreover, there was no significant difference in mean VOR gain in total darkness between the two groups. Gaze gain of patients (0.26+/-0.06 versus 0.81+/-0.06) but not control subjects (0.91+/ 0.07 versus 0.88+/-0.08), was significantly different during pursuit and VOR cancellation. CONCLUSION: SCA-6 patients show dissociation in the control of gaze tracking during smooth pursuit and VOR cancellation. PMID- 10690978 TI - Neuropsychological and clinical correlates of antisaccade task performance in schizophrenia. AB - OBJECTIVES: To elucidate pathophysiologic mechanisms involved in abnormal antisaccade task performance in schizophrenia by investigating a possible relationship among antisaccade task performance, neuropsychological test results, and symptomatology in a group of young patients with recent-onset schizophrenia; to compare the effects of olanzapine and risperidone on antisaccades and reflexive saccades. BACKGROUND: Patients with schizophrenia consistently perform worse than controls on the antisaccade task in which the subject is required to inhibit a reflexive saccade to a suddenly appearing visual target and look in the opposite direction. METHODS: In 37 young (mean age 21 years), medicated patients with recent-onset schizophrenia the authors assessed antisaccades, reflexive saccades, neuropsychological test performance, and symptomatology. A subgroup of 18 patients was treated with olanzapine, and 15 patients were treated with risperidone. Reflexive-saccade and antisaccade task results were compared with those obtained in 13 control subjects. RESULTS: The antisaccade error rate was significantly higher in the patients than in the control subjects. In the patients, poor working memory function was related to increased antisaccade error rate. Severity of disorganization symptoms at intake was related to prolonged mean latency of the correct antisaccades. Patients on risperidone had a prolonged mean latency in the reflexive saccade task compared with patients using olanzapine. CONCLUSIONS: Abnormal antisaccade task performance is already present in early schizophrenia and may reflect working memory dysfunction. In future studies, medication effects should be considered in interpreting eye movement test results of patients with schizophrenia. PMID- 10690979 TI - Randomized pilot trial of postoperative aspirin in subarachnoid hemorrhage. AB - OBJECTIVE: To assess the safety and feasibility of a clinical trial on the effectiveness of acetylsalicylic acid (ASA) in subarachnoid hemorrhage (SAH). BACKGROUND: Several studies have indicated that increased platelet activity might be involved in the pathogenesis of delayed cerebral ischemia (DCI) after SAH. METHOD: Fifty patients who had early surgery (< or =4 days) for a ruptured aneurysm were enrolled in this randomized, double-blind, placebo-controlled trial. Trial medication, consisting of suppositories with 100 mg ASA versus placebo, was started immediately after surgical clipping of the aneurysm and continued for 21 days. End points were functional outcome and quality of life at 4 months, clinical deterioration after operation, development of DCI, hypodense lesion on postoperative CT, and hemorrhagic complications. RESULTS: One-third of all patients with aneurysmal SAH were eligible for the trial. Fifteen of 26 patients receiving placebo deteriorated clinically versus 10 of 24 patients receiving ASA; 4 patients in each group deteriorated from DCI. Postoperative hypodensities on CT were observed in 27 patients, distributed equally in both groups. Functional outcome and quality-of-life scores were slightly in favor of patients who had received ASA, but not to a significant degree (p = 0.22). Two patients in the ASA group had an asymptomatic hemorrhagic complication, and one patient in the placebo group had a fatal and another a symptomatic hemorrhagic complication. CONCLUSION: This pilot study shows that a clinical trial of acetylsalicylic acid (ASA) in subarachnoid hemorrhage (SAH) is feasible and probably safe. The effectiveness of ASA on functional outcome and delayed cerebral ischemia has to be studied in a larger trial. PMID- 10690980 TI - Benign prognosis of never-symptomatic carotid occlusion. AB - OBJECTIVE: To determine the prognosis of asymptomatic carotid artery occlusion. BACKGROUND: As opposed to symptomatic carotid occlusion, little information is available on the prognosis of asymptomatic carotid occlusion. METHOD: Thirty never-symptomatic and 81 symptomatic patients with carotid occlusion underwent baseline assessment of 15 risk factors together with PET measurements of oxygen extraction fraction (OEF). Every 6-month telephone contact recorded interval medical treatment and subsequent stroke occurrence during an average follow-up of 32 months. Patients, treating physicians, and an end point adjudicator were blinded to PET results. RESULTS: Ischemic stroke occurred in 1 of 30 of never symptomatic patients (3.3%) and 15 of 81 of symptomatic patients (18.5%; p = 0.03). No strokes in the carotid territory distal to the occluded vessel occurred in the never-symptomatic patients. Multivariate analysis of baseline risk factors for all 111 patients revealed that age, plasma fibrinogen level, and PET findings of high OEF distal to the occluded carotid artery were the only independent predictors of subsequent stroke (p < 0.05). Previous ipsilateral hemispheric or retinal symptoms was not a significant predictive variable. The lower risk of stroke in never-symptomatic patients was associated with a lower incidence of high OEF (4 of 30) as opposed to symptomatic patients (39 of 81; p = 0.002), but there was no significant difference in age or fibrinogen level. CONCLUSIONS: Never-symptomatic carotid occlusion carries a very low risk of subsequent ischemic stroke. This benign prognosis is associated with a low incidence of cerebral hemodynamic compromise in these patients. These data support further the importance of hemodynamic factors in the pathogenesis of ischemic stroke in patients with carotid occlusion. PMID- 10690981 TI - Early lacunar strokes complicating polyarteritis nodosa: thrombotic microangiopathy. AB - OBJECTIVE: To determine the patterns and mechanisms of polyarteritis nodosa (PAN) associated strokes (PANAS). BACKGROUND: Strokes are reputed to be rare complications of PAN and to occur at a late stage (2 to 3 years). The cause of stroke is unknown but may be related either to atherosclerosis-like occlusive vasculopathy, caused possibly by hypertension or corticosteroid (CS) use, or to vasculitic arterial occlusion. METHODS: Clinical and radiologic patterns, latencies, and current therapy at onset in 15 PANAS patients (4 of the authors' and 11 published cases) were analyzed. RESULTS: A lacunar stroke syndrome (11/15 cases, 73%) was the most frequent stroke pattern in PANAS (multiple, small, deep infarcts in 6, [55%], pontine lacunae in 3 [27%], and leukoaraiosis in 2 [18%]), followed by pure lobar hematoma and bilateral, possibly cardioembolic, large ischemic infarcts (2 cases each). A stroke latency shorter than that previously established (within 8 months in 73% of cases; mean latency, 6.5 months) and a close relationship between the use of CS and stroke in PAN also were found. Of the 77% of first-time or recurrent lacunar strokes that developed despite CS therapy, 80% appeared within 6 months and 50% within 3 weeks of CS initiation. CONCLUSION: Early lacunar stroke syndrome, related to deep small- or pontine penetrating artery thrombotic microangiopathy rather than vasculitis, was the most frequent PANAS pattern. This vasculopathy may be aggravated by corticosteroid (CS) therapy enhancement of either platelet thromboxane A2 production or arterial wall fibrosis. Thus, antiplatelet drugs in association with CS may be advisable for preventing stroke occurrence or recurrence in PAN. PMID- 10690982 TI - Somatosensory potentials, CSF creatine kinase BB activity, and awakening after cardiac arrest. AB - OBJECTIVE: To examine the utility of somatosensory evoked potential (SEP) peaks and CSF creatine kinase BB isoenzyme activity (CKBB) in predicting nonawakening from coma due to cardiac arrest. BACKGROUND: Accurate predictors of neurologic outcome in patients comatose after cardiac arrest are needed to improve medical decision making. METHODS: A total of 72 comatose patients had bilateral median SEPs, and of these, 52 had CSF and CKBB. Awakening was defined as following commands or having comprehensible speech. Both short (N1) and long (N3) latency SEP peaks were analyzed. Nonparametric analyses were used. RESULTS: For patients who had both tests, CKBB > or = 205 U/L predicted nonawakening with a sensitivity of 49% and a specificity of 100%. Bilateral absence of the N1 peak predicted nonawakening with a sensitivity of 53% and a specificity of 100%. Using CKBB > or = 205 U/L, bilaterally absent SEP N1 peaks, or both predicted nonawakening with a sensitivity of 69% and a specificity of 100%. Using CKBB > or = 205 U/L, bilaterally absent N1 peaks, bilateral N3 > or = 176 msec or absent, or some combination predicted nonawakening with a sensitivity of 78% and a specificity of 100%. CONCLUSION: The combination of an absent N1 peak and elevated CKBB performs better than either alone in predicting nonawakening after cardiac arrest. Prolonged or absent N3 latency may increase sensitivity. These results should be interpreted with caution given the small number of patients and the possibility of a self-fulfilling prophecy. PMID- 10690983 TI - Neuropsychological effects of valproate in traumatic brain injury: a randomized trial. AB - OBJECTIVES: To examine the neuropsychological side effects of valproate (VPA) given to prevent posttraumatic seizures. METHODS: In a randomized, double-masked, parallel group clinical trial, we compared the seizure prevention and neuropsychological effects of 1 or 6 months of VPA to 1 week of phenytoin. We studied 279 adult subjects who were randomized within 24 hours of injury and examined with a battery of neuropsychological measures at 1, 6, and 12 months after injury. We examined drug effects cross-sectionally at 1, 6, and 12 months and longitudinally by examining differential change from 1 to 6 months and from 6 to 12 months as a function of protocol-dictated changes in treatment. RESULTS: No significant adverse or beneficial neuropsychological effects of VPA were detected. CONCLUSIONS: Valproate (VPA) appears to have a benign neuropsychological side effects profile, making it a cognitively safe antiepileptic drug to use for controlling established seizures or stabilizing mood. However, based on this study, VPA should not be used for prophylaxis of posttraumatic seizures because it does not prevent posttraumatic seizures, there was a trend toward more deaths in the VPA groups, and it did not have positive effects on cognition. PMID- 10690984 TI - Is geomagnetic activity a risk factor for sudden unexplained death in epilepsies? AB - OBJECTIVE: To test the hypothesis that sudden unexplained death (SUD) in epilepsy is related to geomagnetic activity. BACKGROUND: Prior studies presume that geomagnetic activity (with average amplitudes above 50 nanotesla [nT]) is associated with SUD in epileptic human patients and in epileptic laboratory rats. METHODS: In a retrospective study, 46 epileptic patients with definite SUD were compared with 108 epileptic patients with known cause of death (KCD) who died between 1981 and 1992. A complete postmortem examination was performed in all cases. The time of the day and date of death, as well as two international geomagnetic indices concerning Bartels' planetary 3-hour signs (Kp) and the mean planetary daily amplitudes (Ap) at time of death, were assessed. RESULTS: Among 45 SUD individuals, the local time (37.8%) and the universal time of death (35.6%) peaked within the critical period between 3 to 9 AM. However, the SUD and KCD group did not substantially differ in regard to the distribution of local or universal time of death (p > 0.2, Fisher test). Neither the Kp signs at death and 2 hours before death nor the Ap values showed considerable differences between the SUD and KCD series (p > 0.2, Mann-Whitney test). Merely 4.3% of SUD patients and 3.7% of KCD patients were associated with Ap indices above 50 nT (p > 0.2, Fisher test). CONCLUSION: The results do not support the hypothesis that geomagnetic activity is related to occurrence of sudden unexplained death in epileptic patients. PMID- 10690985 TI - Bilateral frontal polymicrogyria: a newly recognized brain malformation syndrome. AB - BACKGROUND AND OBJECTIVE: Polymicrogyria is a brain malformation characterized by abnormal cortical lamination, excessive cortical folding, and fusion of the cortical molecular layer. Two distinct bilateral localized forms have been described: bilateral perisylvian polymicrogyria, which has proved to be genetically heterogeneous, and bilateral parasagittal parieto-occipital polymicrogyria, which has been described only in sporadic patients. We describe 13 patients with symmetric polymicrogyria of both frontal lobes back to the precentral sulcus: bilateral frontal polymicrogyria (BFP). METHODS: Review of clinical records, brain MRI, and EEG results of 13 patients; correlation with other regional polymicrogyrias. RESULTS: The abnormal cortex extended from the frontal poles anteriorly to the precentral gyrus posteriorly and to the frontal operculum inferiorly and was relatively symmetric in all 13 patients. All patients presented with developmental delay and mild spastic quadriparesis, but variably impaired language development (12/13), mental retardation (11/13), and epilepsy (5/13) also occurred. BFP was sporadic in 13 of 13 patients, but 2 of 13 had consanguineous parents. CONCLUSIONS: BFP extends the spectrum of the recognized bilateral symmetric regional polymicrogyria syndromes. PMID- 10690986 TI - Brain volume in children with neurofibromatosis type 1: relation to neuropsychological status. AB - OBJECTIVE: To determine characteristics of brain morphology in children and adolescents with neurofibromatosis type 1 and relate these characteristics to neuropsychological functioning. BACKGROUND: Neurofibromatosis type 1 is associated with numerous CNS abnormalities and cognitive impairment. Abnormal high signal intensity visible on brain MRI, brain tumors, and macrocephaly are common. Research into links between neuroanatomic and cognitive features has been inconclusive. METHODS: Fifty-two children and adolescents with neurofibromatosis type 1 were compared with 19 control subjects on several quantitative neuroanatomic and neuropsychological measures. RESULTS: Total brain volume, especially gray matter, was significantly greater for neurofibromatosis type 1 subjects than the control subjects. Group differences in the ratio of gray matter to white matter were more prominent in younger than in older subjects. Volume of gray matter in the subjects with neurofibromatosis type 1 was related to their degree of learning disability. Corpus callosum size was significantly larger for subjects in the neurofibromatosis type 1 group, and diminished performance on measures of academic achievement and visual-spatial and motor skills were associated with greater regional corpus callosum size. CONCLUSIONS: Neuroanatomic morphology and the developmental pattern of gray matter and white matter in subjects with neurofibromatosis type 1 differed from in control subjects. Some of these differences are related to the neuropsychological status of the neurofibromatosis type 1 group. We propose that delayed developmental apoptosis results in macrocephaly and a delay in the development of appropriate neuronal connections in children with neurofibromatosis type 1. We further propose that these morphologic delays are related to the cognitive profile of neurofibromatosis type 1. PMID- 10690987 TI - Cerebrovascular changes in the basal ganglia with HIV dementia. AB - BACKGROUND: HIV dementia is a form of subcortical dementia. Clinical, radiologic, pathologic, and biochemical studies suggest a major contribution of basal ganglia dysfunction to the pathogenesis of this disorder. Many investigators have proposed a contribution of a disrupted blood-brain barrier (BBB) to the pathogenesis of HIV dementia. OBJECTIVE: To identify microvascular abnormalities in vivo in basal ganglia or white matter of persons with HIV dementia. METHODS: Time course of MRI postcontrast enhancement was determined in basal ganglia and white matter of HIV-infected persons without dementia (Memorial Sloan Kettering [MSK] score of 0; n = 4); HIV-infected persons with mild dementia (MSK score of 0.5; n = 2); and HIV-infected persons with moderate-to-severe dementia (MSK > or = 1.0; n = 6). RESULTS: Increased basal ganglia enhancement was observed in individuals with moderate-to-severe dementia relative to nondemented individuals, both immediately and 30 minutes after contrast administration. Decline of basal ganglia enhancement was slower in the moderately to severely demented patients and, when normalized to intravascular enhancement of sagittal sinus, suggested leakage of contrast agent, consistent with increased permeability of BBB. A significant correlation between the postcontrast fractional enhancement at 30 minutes (FE30) and the MSK score was noted. White matter showed no significant differences in postcontrast enhancement among the three groups. CONCLUSION: Increased early enhancement in basal ganglia of the HIV dementia group is consistent with increased regional cerebral blood volume (rCBV). Increased late enhancement is strongly suggestive of BBB disruption. Similar abnormalities were absent in the white matter adjacent to the caudate nucleus. PMID- 10690988 TI - Cerebrospinal fluid HIV RNA originates from both local CNS and systemic sources. AB - OBJECTIVE: To identify the sources of HIV virions in CSF by modeling treatment associated HIV dynamics. BACKGROUND: We postulated a model in which cell-free CSF virions originate from two major sources, namely, systemic non-CNS and CNS tissues, the latter including brain parenchyma and meninges. The model predicted that with initiation of antiretroviral therapy, the acute-phase decline in CSF HIV RNA levels would be controlled by the kinetics of the dominant virion source (systemic versus CNS). Based on prior observations, we hypothesized that the dominant source of CSF virions would shift from systemic to CNS in more advanced disease. METHODS: Three patient groups were studied: Group 1 (n = 5): nondemented, with early HIV disease (CD4+ lymphocytes > or = 400/microL) or pleocytosis (CSF leukocytes > or = 4/microL); Group 2 (n = 5): nondemented, with advanced HIV disease (CD4+ < 400/microL) and no pleocytosis; Group 3 (n = 2): patients with HIV-associated dementia (HAD). All patients began a new, highly active antiretroviral treatment regimen and underwent serial lumbar punctures and phlebotomies. RESULTS: For patients in Group 2, the rate of decline in CSF HIV RNA was slower than in plasma (p < 0.00001). For Group 1, the rate of decline in CSF was not different from plasma (p > 0.25). Patients with HAD showed high CSF HIV RNA after 5 to 6 weeks of treatment despite a 100-fold decrease in plasma HIV RNA. CONCLUSIONS: CSF and plasma HIV dynamics became increasingly independent in advanced HIV disease, and the compartmental discrepancy was largest in HAD. Our findings suggest that viral replication in CNS tissues may constitute a major, independent source of CSF HIV RNA. In patients with HAD, brain parenchyma itself may be the principal CNS tissue source, and CNS-targeted treatment strategies may be required to eradicate this infection. PMID- 10690989 TI - Functional consequences of chloride channel gene (CLCN1) mutations causing myotonia congenita. AB - OBJECTIVE: To determine the functional consequences of missense mutations within the skeletal muscle chloride channel gene CLCN1 that cause myotonia congenita. BACKGROUND: Myotonia congenita is a genetic muscle disease associated with abnormalities in the skeletal muscle voltage-gated chloride (ClC-1) channel. In order to understand the molecular basis of this inherited disease, it is important to determine the physiologic consequences of mutations found in patients affected by it. METHODS: The authors used a mammalian cell (human embryonic kidney 293) expression system and the whole-cell voltage-clamp technique to functionally express and physiologically characterize five CLCN1 mutations. RESULTS: The I329T mutation shifted the voltage dependence of open probability of ClC-1 channels to the right by 192 mV, and the R338Q mutation shifted it to the right by 38 mV. In addition, the I329T ClC-1 channels deactivated to a lesser extent than normal at negative potentials. The V165G, F167L, and F413C ClC-1 channels also shifted the voltage dependence of open probability, but only by +14 to +20 mV. CONCLUSIONS: The functional consequences of these mutations form the physiologic argument that these are disease-causing mutations and could lead to myotonia congenita by impairing the ability of the skeletal muscle voltage-gated chloride channels to maintain normal muscle excitability. Understanding of genetic and physiologic defects may ultimately lead to better diagnosis and treatment of patients with myotonia congenita. PMID- 10690990 TI - Psychometric evaluation of a new sensory scale in immune-mediated polyneuropathies. Inflammatory Neuropathy Cause and Treatment (INCAT) Group. AB - OBJECTIVE: To perform a psychometric evaluation of the inflammatory neuropathy cause and treatment (INCAT) sensory sumscore (ISS) in sensory-motor immune mediated polyneuropathies. This new sensory scale was evaluated to strive for uniformity in assessing sensory deficit in these disorders. METHODS: The ISS comprises vibration and pinprick sense plus a two-point discrimination value and ranges from 0 (normal sensation) to 20 (maximum sensory deficit). Before its clinical use, a panel of expert neurologists concluded that the ISS has face and content validity. The construct validity of the ISS was investigated by correlation and regression studies with additional scales (Nine-Hole Peg Test, 10 Meter Walking Test, a disability sumscore). All scales were applied in 113 patients with a stable neurologic condition (83 patients who experienced Guillain Barre syndrome [GBS] in the past, 22 with chronic inflammatory demyelinating polyneuropathy [CIDP], 8 patients with a monoclonal gammopathy associated polyneuropathy), and 10 patients with recently diagnosed GBS or CIDP with changing clinical conditions. Reliability of the ISS was evaluated in the stable patients. Its responsiveness was investigated in the patients examined longitudinally. RESULTS: A moderate to good validity was obtained for the ISS (stable group: r = 0.38 to 0.56, p < or = 0.006; longitudinal group: R = 0.60 to 0.82, p < or = 0.007, except for the association with the 10-Meter Walking Test [p = 0.08]). Acceptable internal consistency, and inter- and intraobserver reliability were demonstrated for the ISS (alpha = 0.68 to 0.87; R = 0.85 to 0.89, p < 0.0001). Standardized response mean scores for the ISS were high (> or =0.8), indicating good responsiveness. CONCLUSIONS: All psychometric requirements are provided for the the inflammatory neuropathy cause and treatment sensory sumscore. The use of this scale is therefore suggested for bedside evaluation of sensory deficit in the individual patient with a sensory-motor immune-mediated polyneuropathy as well as in clinical trials. PMID- 10690991 TI - Molecular and clinical analyses of spinocerebellar ataxia type 8 in Japan. AB - OBJECTIVE: To clarify the molecular and clinical features of the newly identified spinocerebellar ataxia type 8 (SCA8). METHODS: We analyzed the CTG repeat region of the SCA8 gene in a series of Japanese patients with cerebellar ataxia. We also investigated the frequency of the CTG repeat length in Japanese normal elderly subjects older than age 79. Morphometric measurements on the cerebral MRI were compared between patients with SCA8 and SCA6. RESULTS: The number of the combined CTA/CTG repeats of six affected SCA8 alleles was 106.3+/-24.4 (mean +/- SD) ranging from 89 to 155 and that of normal elderly subjects was 24.3+/-4.4 (n = 104 alleles) ranging from 15 to 34. The mean age at onset of the SCA8 cases was 53.8+/-19.7 years, with a range from 20 to 73 years. One father and daughter from an SCA8 family showed remarkable paternal anticipation. The number increase from father to daughter was + 16 CTG repeats, with a 31-year acceleration of onset. The six identified SCA8 patients were clinically characterized by high frequencies of incoordination of trunk and limbs, ataxic dysarthria, impaired smooth pursuit, and horizontal nystagmus, and the MRI showed significant atrophy of the cerebellar vermis and hemispheres compared with that of normal controls. There was no significant difference between SCA8 and SCA6 on the morphometric MRI study. CONCLUSIONS: The CTG repeat expansions in the SCA8 alleles were much greater than the range of repeats in normal elderly subjects. The SCA8 phenotype manifested by cerebellar symptoms and atrophy corresponded to features of the autosomal dominant cerebellar ataxia type III (ADCA III). PMID- 10690992 TI - Lasting cortical activation after repetitive TMS of the motor cortex: a glucose metabolic study. AB - OBJECTIVE: Cerebral [18F]fluorodeoxy-D-glucose PET ([18F]FDG-PET) was used to visualize the lasting neuronal activation after repetitive transcranial magnetic stimulation (rTMS) over the left hand area of the primary motor cortex (M1HAND). BACKGROUND: Applied over M1HAND, rTMS has been shown to produce a modulation of corticomotor excitability beyond the time of stimulation itself. METHODS: Eight right-handed subjects underwent nonquantitative [18F]FDG-PET measurements during two experimental conditions: at rest and after focal subthreshold 5-Hz rTMS over the left M1HAND. In the post-rTMS condition, [18F]FDG was injected immediately after the administration of 1,800 magnetic pulses over the left M1HAND. Relative differences in normalized regional cerebral metabolic rate of glucose (normalized rCMRglc) between conditions were determined using a voxel-by-voxel Student's t test and volume-of-interest (VOI) analysis. Analysis was a priori restricted to the M1HAND, the supplementary motor area (SMA), and the primary auditory cortex of both hemispheres. RESULTS: A 5-Hz rTMS of the left M1HAND caused a lasting relative increase in normalized rCMRglc within the M1HAND bilaterally and the SMA. The magnitude and the topographic pattern of persisting relative rCMRglc increases within these motor cortical areas demonstrated considerable interindividual variations. CONCLUSIONS: Subthreshold 5-Hz repetitive transcranial magnetic stimulation (rTMS) over the hand area of the primary motor cortex is associated with a persisting neuronal activation in a distinct set of motor cortical areas beyond the time of stimulation. The current findings demonstrate that [18F]FDG-PET can localize and quantify regional net changes in synaptic cortical activity after rTMS and thus might elucidate the mechanisms underlying rTMS-associated therapeutic effects. PMID- 10690994 TI - Psychogenic status epilepticus in children: psychiatric and other risk factors. AB - The authors studied six children with repetitive psychogenic seizures severe enough to mimic status epilepticus. All received IV antiepileptic drugs in an emergency setting. Most had a family history of epilepsy. Affective and anxiety disorders predominated as comorbid psychiatric diagnoses. Acutely stressful situations precipitated all episodes of nonepileptic status epilepticus. With aggressive psychotherapeutic intervention and pharmacologic treatment of their underlying psychiatric diagnosis, the patients improved. PMID- 10690993 TI - Involvement of the ventrolateral medulla in parkinsonism with autonomic failure. AB - OBJECTIVE: To determine whether patients with PD and autonomic failure (AF), manifested primarily with orthostatic hypotension (OH), have a consistent loss of tyrosine hydroxylase (TH) neurons in the rostral ventrolateral medulla (RVLM), similar to that occurring in patients with multiple system atrophy (MSA) and AF, and to determine whether there is loss of nicotinamide, adenine dinucleotide phosphate (NADPH) diaphorase (NADPH-d) RVLM neurons in both groups of patients. METHODS: The numbers of TH and NADPH-d neurons in the RVLM was assessed in brain sections obtained at autopsy from five patients with suspected PD and OH, six patients with MSA, two patients with corticobasal ganglionic degeneration and no AF, and 10 control subjects with no history of neurologic disease. Cell numbers were compared among groups and correlated with their final neuropathologic diagnosis. RESULTS: The number of TH neurons in the RVLM of patients with PD and OH were not significantly different from control subjects, and there were marked individual variations. The TH cell numbers in the RVLM were significantly higher (p < 0.06) in patients with PD than in patients with MSA, despite a similar degree of severity of OH. As a group, patients with PD and OH had reduced numbers of NADPH-d cells in the RVLM compared with control subjects, but again there were marked individual variations. NADPH-d cell numbers were reduced consistently and more markedly in patients with MSA. CONCLUSION: Unlike the case in patients with MSA, the number of TH neurons in the RVLM is highly variable in patients with PD and is unlikely to contribute significantly to the pathophysiology of OH. As a group, patients with PD have reduced numbers of NADPH-d neurons in the RVLM, but some patients had cell counts similar to control subjects. On the other hand, NADPH-d cell depletion in the RVLM is a consistent finding in MSA and may contribute to cardiorespiratory dysfunction in this disorder. PMID- 10690995 TI - "Pressure to laugh": an unusual epileptic symptom associated with small hypothalamic hamartomas. AB - Gelastic seizures are the hallmark of the epilepsy syndrome associated with hypothalamic hamartomas. Patients typically develop cognitive deterioration and refractory seizures. The authors describe three patients with small hypothalamic hamartomas without these features and thus identify a mild end to the clinical spectrum. All had the unusual symptom of "pressure to laugh," often without actual laughter. This symptom could be dismissed as psychogenic but should be recognized as a clue to the presence of this unusual lesion. PMID- 10690996 TI - Stable weight during lamotrigine therapy: a review of 32 studies. AB - A side effect associated with the use of some antiepileptic drugs (AEDs) is change in body weight. To evaluate the effect of lamotrigine on body weight in adult patients with epilepsy, we conducted a retrospective review of data from 463 patients treated with lamotrigine in 32 clinical trials. Mean daily dose was 259 (+/-155) mg and duration of therapy was 318 (+/-87) days. The mean change in body weight was 0.5 (+/-5) kg. Lamotrigine was associated with stable body weight in patients with epilepsy. PMID- 10690997 TI - Bilateral focal motor status epilepticus with retained consciousness after stroke. AB - Bilateral motor seizures with retained consciousness are rare and often mistaken for pseudoseizures. In the few reported cases, the seizures were brief and the underlying lesion usually was a tumor. Here the authors describe a patient with bilateral focal motor status epilepticus with retained consciousness after a stroke. A seizure should be considered as the possible cause of continuous bilateral limb movements with retained consciousness. PMID- 10690998 TI - Amnesic confabulatory syndrome after focal basal forebrain damage. AB - A 73-year-old woman developed amnesic confabulatory syndrome after a right focal basal forebrain hemorrhage. The confabulation, despite persistent antegrade amnesia, gradually subsided with improvement of the frontal executive function. The lesion appeared to disrupt connections of the medial and lateral limbic circuits important for memory. Simultaneous dysfunctioning of the two circuits involving the medial temporal and frontal lobes may be necessary for the development of this syndrome. PMID- 10690999 TI - Relationship between motor and language activation using fMRI. AB - The authors examined laterality ratios (i.e., [L-R]/[L+R]) from functional MRI (fMRI) scans obtained in 12 healthy volunteers during unimanual left- and right hand finger movements and during a verb generation language task. The language and right-hand motor asymmetry ratios were correlated (rho = 0.71, p = 0.005) as were the left- and right-hand ratios (rho = -0.68, p = 0.008). Subjects with greater relative left hemisphere lateralization of language exhibit greater relative unilateral hemisphere activation during right-hand movements. PMID- 10691000 TI - Residual function in motor cortex contralateral to amputated hand. AB - OBJECTIVE: To investigate residual function of the motor cortex corresponding to the hand of the amputated arm (MCamp). METHODS: Focal transcranial magnetic stimulation (TMS) of MCamp was performed in 10 patients 22 to 52 years after arm amputation to inhibit tonic muscle contraction in the intact hand ipsilateral to cortex stimulation. RESULTS: In all patients, onset latency, degree, and duration of this inhibition were normal. CONCLUSION: The presence of motor inhibition in the residual hand of amputees originating from the hand motor representation of MCamp indicates residual cortical motor representation of the lost hand irrespective of whether the effect is mediated by commissural or ipsilateral corticospinal connections. PMID- 10691001 TI - Cerebral CO2 reactivity, cholesterol, and high-density lipoprotein cholesterol in the elderly. AB - Cholesterol and its subfractions play a role in the development of atherosclerosis. Cerebral CO2 reactivity reflects the compensatory capacity of cerebral arterioles. The authors investigated the relationship between total cholesterol, high-density lipoprotein (HDL), their ratio, and cerebral CO2 reactivity in 826 participants from the Rotterdam Study. Cerebral CO2 reactivity increased significantly with increasing levels of HDL and decreased significantly with an increasing total cholesterol/HDL ratio. This suggests that blood lipids may also affect smaller cerebral blood vessels. PMID- 10691002 TI - Language disturbances in corticobasal degeneration. AB - To characterize the language deficits in corticobasal degeneration (CBD) and their relation to neuroradiologic findings, the authors administered a standardized battery of neurobehavioral tests and performed MRI studies on 15 patients with CBD. Eight patients (53%) had classifiable aphasias, including anomic, Broca's and transcortical motor aphasias. The aphasias were associated primarily with left frontal and parietal cortical damage and subcortical white matter and corpus callosum abnormalities. Our findings demonstrate that language disturbances in CBD are more frequent than previously considered. PMID- 10691003 TI - Role of brain biopsy in the management of focal brain lesions in HIV-infected patients. Gruppo Italiano Cooperativo AIDS & Tumori. AB - In this multicenter, retrospective study of 160 brain biopsies in the assessment of HIV-related focal brain lesions, diagnostic sensitivity was acceptable (87%), but the procedure carried considerable morbidity (7.5%) and mortality (3.1%). Moreover, it is not always possible to initiate the changes in therapy indicated by the results, and overall survival remains poor, with a median of 2 months. Criteria for brain biopsy for the diagnosis of focal brain lesions should be redefined to include selected patients for whom a less invasive approach does not yield a definitive diagnosis. PMID- 10691004 TI - Longitudinal study of neurotoxicity with occupational exposure to aluminum dust. AB - Two cross-sectional studies were conducted at a German aluminum (Al) powder plant to evaluate possible nervous system effects from occupational Al exposure. The investigation included biological monitoring, a neuropsychological test battery, and event-related P300 potentials. Neurophysiologic findings in workers chronically exposed to Al dust did not differ from non-Al-exposed controls from the same plant. The authors suggest that chronic exposure to Al dust, at the levels documented in this study, does not induce measurable cognitive decline. PMID- 10691005 TI - Chronic ophthalmoplegia with anti-GQ1b antibody. AB - Anti-GQ1b antibodies are typically found in patients with the Miller Fisher syndrome, all of whom will have, by definition, acute ophthalmoplegia. The authors describe three patients with chronic ophthalmoplegia in the presence of persistently high titers of immunoglobulin G anti-GQ1b antibody detected in an ELISA, one of whom improved with immunotherapy. Anti-GQ1b antibodies may be associated with some cases of chronic ophthalmoplegia of unknown cause. PMID- 10691006 TI - Potential role of LIF as a modifier gene in the pathogenesis of amyotrophic lateral sclerosis. AB - Leukemia inhibitory factor (lif) is a potent survival factor for motoneurons in cell culture and in vivo. The authors screened 104 patients with ALS and 338 control subjects for mutations in the LIF gene. In four ALS patients, but in no control subject, a G-to-A point mutation at position 3400 was identified, which leads to an amino acid exchange of valine to methionine at position 64 of the mature lif protein. This region of the lif protein (AB loop) interacts with the lif receptor. The authors suggest that LIF could act as a modifier gene which, in combination with other genetic predispositions, might lead to motoneuron disease. PMID- 10691007 TI - mtDNA A3243G MELAS mutation is not associated with multigenerational female migraine. AB - The authors searched for mitochondrial DNA (mtDNA) A3243G mutation in peripheral blood leukocytes from female migraine patients with pure matrilinear history of migraine along two or three generations. The current study was designed to exclude any male transmission of the disease. The mutation was absent in all patients. We conclude that mtDNA A3243G mutation does not contribute to the pathogenesis of pure matrilinear multigenerational migraine with or without aura. PMID- 10691008 TI - A successful pregnancy in a heterozygote for OTC deficiency treated with sodium phenylbutyrate. PMID- 10691009 TI - Immunoglobulin therapy for idiopathic chronic sensory ataxic neuropathy. PMID- 10691010 TI - Stroke treatment with tissue plasminogen activator in the setting of aortic dissection. PMID- 10691011 TI - Delayed myelopathic presentation of the acquired hepatocerebral degeneration syndrome. PMID- 10691012 TI - Cerebellar degeneration associated with HIV infection. PMID- 10691013 TI - Acute deterioration from thrombosis and rerupture of a giant intracranial aneurysm. PMID- 10691014 TI - Charcot-Marie-Tooth disease type 2 with restless legs syndrome. PMID- 10691015 TI - Dementia is a major predictor of death among the Italian elderly. PMID- 10691016 TI - Contrast agent neurotoxicity presenting as subarachnoid hemorrhage. PMID- 10691017 TI - Detecting drug effects on short-term memory function using a combined delayed matching and non-matching to position task. AB - Operant delayed non-matching-to-position (DNMTP) and delayed matching-to-position (DMTP) have become standard techniques to investigate drug effects on short-term memory function in rats. However, these two tasks are normally conducted in isolation. Using two standard drugs, the 5HT1A agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), and the muscarinic antagonist scopolamine, this study looked at a two-choice operant task that essentially involved a mixed DNMTP/DMTP paradigm. Thus, DNMTP trials were interspersed with DMTP trials in a random sequence for the duration of a session. 8-OH-DPAT (0.03 mg/kg) slightly but significantly improved response accuracy in a delay-dependent fashion during DMTP but not DNMTP trials. The highest dose of 8-OH-DPAT (0.1 mg/kg) impaired accuracy during DNMTP trials independent of delay and had no significant effect during DMTP trials. Scopolamine (0.1 mg/kg) produced delay-dependent deficits in accuracy during DMTP trials but delay-independent impairments during DNMTP trials. Because both 8-OH-DPAT and scopolamine produced delay-dependent effects with DMTP trials types and either had no effect (8-OH-DPAT) or produced delay independent impairments (scopolamine) during DNMTP trials types, it is suggested that DMTP trials had a greater dependence on short-term working memory function than DNMTP trials that probably relied more on positional (mediating) strategies for solving the task. Therefore, we believe that this mixed DNMTP/DMTP task offers greater potential for more reliable and discerning interpretation of data regarding short-term memory function in rodents than either of the paradigms performed in isolation. PMID- 10691018 TI - Analysis of dextrorphan, a metabolite of dextromethorphan, using gas chromatography with electron-capture detection. AB - Dextromethorphan, a constituent of many over-the-counter cough syrups, is used as a probe drug for phenotyping subjects for their cytochrome P450 2D6 (CYP2D6) enzyme activity and for measuring CYP2D6 activity of preparations such as microsomes. In such studies, formation of the metabolite dextrorphan is used as indicator of the activity of this CYP enzyme. The present report describes an electron-capture gas chromatographic procedure developed for detection and quantification of dextrorphan in human liver microsomal preparations in vitro. After basification of the incubation mixture, dextrorphan was derivatized with pentafluorobenzoyl chloride under aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column, an electron capture detector, and a printer-integrator. Para-hydroxymephenytoin was carried through the procedure as internal standard. The procedure, which involves the derivatization of dextrorphan under aqueous conditions, is rapid and involves the use of the relatively economical procedure of electron-capture gas chromatography. The derivative is stable and possesses excellent chromatographic properties. PMID- 10691019 TI - Evaluation of erectile response by continuous measurement of penile diameter in rats. AB - The present study was performed to determine whether measurement of penile diameter in an in vivo rat model is useful for pharmacologic and physiologic investigations on penile erection. Penile erection induced by electrical stimulation of the cavernous nerve was monitored by measuring the penile diameter sonomicrometrically with a pair of 10-MHz piezoelectric crystals glued to the opposite surfaces of the adventitia of the penile erectile chamber in anesthetized rats. Using this method, we examined the effects of a nitric oxide (NO) synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME), and a well-known phosphodiesterase 5 (PDE5) inhibitor, zaprinast, on the maximal developed penile diameter (D-max) and the time from the maximum response to 50% recovery (T50%) of the maximum response as an index of the duration of penile erection. An intravenous injection of L-NAME at a dose of 10 mg/kg significantly inhibited D-max produced by cavernous electrical stimulation at 5 to 50 V, without affecting T50%. Sequential intravenous infusions of 10, 30, 100, and 300 microg/kg/min of zaprinast at 30-min intervals did not show any effect on D-max, heart rate, and systolic arterial pressure, although doses of 100 and 300 microg/kg/min significantly prolonged T50% and the maximum dose decreased diastolic arterial pressure. Moreover, zaprinast produced a more prominent increase in cyclic guanosine monophosphate (cGMP) levels than cyclic adenosine monophosphate levels in the plasma taken at the end of the maximum dose infusion. Measurement of murine penile diameter with a sonomicrometrical device, indicating that a NO-cGMP-PDE5 pathway plays a pivotal role in the penile diameter increase and its maintenance, would be useful for pharmacologic and physiologic investigations on penile erection. PMID- 10691020 TI - Methodologic aspects of acetylcholine-evoked relaxation of rabbit aorta. AB - The acetylcholine-evoked relaxation of rabbit isolated thoracic aorta precontracted by phenylephrine was studied. Phenylephrine caused a steady contraction that was maintained for 6 h. In the presence of calcium disodium ethylenediaminetetraacetate (EDTA) and ascorbic acid the contraction decreased with time. N(G)-Nitro-L-arginine abolished the inhibitory effect of EDTA and ascorbic acid. Acetylcholine evoked a rapid concentration-dependent relaxation that recovered spontaneously and slowly, but fully, with time. Relaxation evoked by equieffective concentrations of carbachol and acetylcholine had the same time course. Cumulative addition of acetylcholine (10(-7)-3 x 10(-5) M) caused a marked relaxation that was reverted slightly at high concentrations. The relaxation was the same with rings derived from the upper, middle, and lower part of the thoracic aorta. Two consecutive concentration-response curves for acetylcholine obtained at a 2-h interval demonstrated a slight development of tachyphylaxis. The relaxation was inversely related to precontractile tension evoked by phenylephrine when expressed as a percentage, but independent when expressed as g tension. Storage of aorta in cold salt solution for 24 h did not alter the relaxation. EDTA and ascorbic acid did not alter the relaxation. It is concluded that (1) EDTA and ascorbic acid can not be used with impunity to stabilize catecholamines used as preconstriction agents; (2) the reversal of the acetylcholine-evoked relaxation is not due to hydrolysis of acetylcholine; (3) the relaxation is uniform in all segments of thoracic aorta; (4) cold storage of aorta does not alter the relaxation; and (5) acetylcholine releases the same amount of relaxing factor, irrespective of the precontractile tension. PMID- 10691021 TI - Wall stress-induced dysrhythmias in the isolated working rat heart perfused through a cannula placed in the left ventricle via aorta. AB - The purpose of the present study was to determine if our recently introduced novel working rat heart preparation could be used to study wall stress-induced dysrhythmias. A double cannula, which consisted of an outer cannula that, was inserted in the aorta and an inner cannula that was advanced into the left ventricle was used. The perfusion flowed through the inner cannula into the left ventricle and was ejected from there into the aorta. Afterload was changed suddenly from 60 to 160 Hg of pressure by turning a valve so that the fluid was diverted to a column set at a different height. A sudden increase of aortic pressure that lasted for 10 sec caused cardiac ectopic beats. Wall stress-induced dysrhythmias were more sustained during perfusion with low potassium and low magnesium Krebs-Henseleit solution. Bradykinin (1 microg) or epinephrine (10 microg) was injected as a bolus via an in-line injection port placed at the inner cannula. Bradykinin significantly reduced the incidence of ectopic beats and epinephrine increased the incidence of nonsustained runs of VT. This "working" heart preparation is a convenient tool to study wall stress-induced dysrhythmias. PMID- 10691022 TI - A new in vitro model for ethanol-induced gastric mucosal damage. AB - A new in vitro model for ethanol-induced gastric damage is described. The stomach was dissected from the rat, a polyethylene cannula introduced into the remnants of the esophagus, and the pyloric end tied off. With the cardiac and pyloric regions of the stomach secured by thread to a vertical glass rod or tube, the whole was suspended in an organ bath containing aerated Krebs solution. Fifteen minutes later, ethanol was introduced via the esophageal cannula. After an additional 60 min, the stomach was removed from the Krebs solution, opened along the mid line, and the lesions studied. Comparisons were made with a conventional in vivo model. Results show that the lesion number, length, and total lesion area obtained by the in vitro model were comparable to those obtained in the older in vivo model. Histopathologically, lesions induced by both models were also comparable. Clonazepam, a drug previously used in the in vivo model, was tested in this model. Results indicate that clonazpam protected against ethanol-induced gastric damage in vitro. The new model provides a method to study the action of drugs on the stomach alone and to exclude in indirect actions of drugs via other sites in the body. PMID- 10691023 TI - The mechanism of thrombin-induced prostacyclin synthesis in human endothelial cells with reference to the gene transcription of prostacyclin-related enzymes and Ca2+ kinetics. AB - This study was designed to evaluate the effect of thrombin on prostacyclin (PGI2) production in cultured human vascular endothelial cells in association with intracellular Ca2+ and with the gene expression of prostaglandin H2 synthase (PGHS) and phospholipase A2 (PLA2) using competitive polymerase chain reaction. Thrombin enhanced the PGI2 synthesis dependent with time. Additionally, thrombin increased the intracellular Ca2+, which stimulates PLA2, resulting in arachidonic acid cleavage from membrane phospholipids and its subsequent conversion into PGI2 through the PGHS pathway. The elevation of intracellular Ca2+ was a result of Ca2+ influx and Ca2+ release from its intracellular storage sites. In this study, PGHS-1 mRNA was constitutively expressed, whereas PGHS-2 mRNA was not. With the stimulation of thrombin, cytosolic PLA2 (cPLA2) mRNA increased 9-fold at 15 min, PGHS-1 mRNA increased 3.4-fold at 180 min, and PGHS-2 mRNA increased 38-fold at 60 min. These results suggest that the elevation of intracellular Ca2+ and the expression of cPLA2, PGHS-1, and PGHS-2 mRNA cause PGI2 generation. PMID- 10691024 TI - A time course study for the development of an immunocompromised wound model, using hydrocortisone. AB - Although wound healing is essentially a physiologic process, some chronic wounds exhibit considerable delay in healing. Often these do not heal perfectly in individuals with low immune profiles. Thus, the present study was undertaken to develop an excision wound model in the immunocompromised state induced by pretreatment with hydrocortisone (HC) 40 mg/kg intramuscularly in male rats. Wounds of 8-mm diameter were made on the preshaved dorsal surface of rats using an Acuderm biopsy punch, following pretreatment with HC. After 14 days HC-treated animals exhibited atrophy of spleen and adrenal glands and a significant reduction of circulating lymphocytes and increase in neutrophils; these changes are indicative of immunosuppressive state of animals. The cell proliferation was significantly affected as shown by decreases in DNA (23%) and protein (11%). Furthermore, there were also significant reductions in tensile strength (37%) and hydroxyproline (33%) contents. These results were further supported by lack of contraction of wound edges. It is concluded that animals primed with HC 1 week prior to wounding developed prolonged immunosuppression, which significantly impaired the wound healing as compared with other groups. Thus, this can be experimentally employed as an immunocompromised wound model for evaluating compounds as novel wound healers suitable for immunocompromised subjects. PMID- 10691025 TI - A new method for recording surface compound potentials in sympathetic ganglia from mouse, rat, and guinea pig--application to muscarinic and nicotinic depolarizations. AB - We present a new method for electrophysiologic investigations in isolated autonomous ganglia of a variety of laboratory animals. This method enables determination of surface compound potentials in ganglia and changes induced by pharmacologic compounds. Advantages of our methods are as following: (1) the method is relatively simple and does not require sophisticated experimental setups, with minor modifications it is adaptable to investigate ganglia of varying sizes; (2) the signal amplitude is comparable or even higher when compared with signals obtained by other methods: (3) the apparatus allows fast addition and removal of the investigational compounds and thus the determination of acute and subacute desensitizing effects; and (4) fast preparation and minor tissue injuries during preparation of the ganglia allow determination of surface potential changes over a time period of up to 2 days without qualitative changes of the parameters. In this report we demonstrate the validity of this method using superior cervical ganglia from rat, mouse, and guinea pig. Agonists used to trigger potential changes are the cholinergic agonists acetylcholine, muscarine, nicotine, and carbachol. The possibility of receptor desensitization by these compounds is investigated by repeated application over 5 h. PMID- 10691026 TI - Inhibition of the action of the topoisomerase II poison amsacrine by simple aniline derivatives: evidence for drug-protein interactions. AB - The action of the anticancer drug amsacrine appears to involve molecular interactions with both DNA and topoisomerase II. It has been shown previously that DNA intercalators can inhibit the action of amsacrine and several other topoisomerase II poisons, presumably as a result of interference with the DNA binding sites for the enzyme. We show here that drug molecules such as N phenylmethanesulfonamide, which mimic the anilino side chain of amsacrine, inhibit the cytotoxicity against cultured Lewis lung murine carcinoma of amsacrine, amsacrine analogues including asulacrine and DACA (N-[2 (dimethylamino)-ethyl]acridine-4-carboxamide dihydrochloride), and etoposide. In contrast, the cytotoxicity of doxorubicin was slightly increased by co-incubation with N-phenylmethanesulfonamide. The cytotoxicity of amsacrine was also modulated in human Jurkat leukemia, HCT-8 colon, and HT-29 colon cell lines. Because o AMSA, an amsacrine analogue containing a methoxy group in the ortho rather than in the meta position, is known to be inactive as an antitumor drug, the abilities of the ortho and meta methoxy-substituted derivatives of methyl-N-phenylcarbamate to reverse the cytotoxicity of amsacrine, asulacrine, and DACA were compared. The ortho substitution decreased activity while meta substitution slightly increased it, suggesting that the side chains were binding to a similar site to that occupied by amsacrine. To determine whether the side chain variants actively inhibited the formation of DNA-topoisomerase II covalent complexes, cultured cells were treated with amsacrine or asulacrine, harvested, and lysed directly on acrylamide gels before electrophoresis and Western blotting to identify non-DNA bound topoisomerase II. Extractable topoisomerase II was depleted in cells incubated with amsacrine but partially restored by coculture with methyl-N phenylcarbamate. The findings are consistent with the hypothesis that low molecular weight molecules can modulate the effects of topoisomerase II poisons by directly interacting with the enzyme. PMID- 10691028 TI - Influence of static magnetic field on the antiproliferative effects of vitamin D on human breast cancer cells. AB - We describe the effect of a 0.2 tesla (T) static magnetic field generated by a magnetic resonance tomograph and of vitamin D treatment on a human breast cancer cell line (MCF-7). Cell damage and proliferation were monitored by measuring the incorporation of [3H]thymidine in duplicating DNA and by the clonogenic assay. [3H]Thymidine incorporation in MCF-7 was stimulated by vitamin D at low doses (10(-12)-10(-10) M), whereas it was inhibited at higher concentrations (10(-9) 10(-6) M). Magnetic field treatment (0.2 T) decreased [3H]thymidine incorporation in human breast cancer cells, eliminating the proproliferative effect of low doses of vitamin D, and enhanced the vitamin D antiproliferative effect, further reducing [3H]thymidine incorporation, from -12.5% (P < 0.05) to -66.7% (P < 0.001), over the range of 10(-9) to 10(-6) M. In the clonogenic assay, ability of MCF-7 to form colonies was inhibited by vitamin D 10(-9) M and above, whereas 3-h exposure to 0.2 T magnetic field had no effect on the number of cell colonies formed. In conclusion, vitamin D treatment yields a permanent antiproliferative effect, while magnetic field exposure only temporarily slows down cellular growth. These findings suggest that therapy with vitamin D may prove beneficial for chemoprevention or treatment of breast cancer. Static magnetic field, alone or in combination, does not appear to represent an effective candidate for breast cancer therapy, at least at the intensity used in the present study. PMID- 10691027 TI - The protective effect of estrogen against chemically induced murine colon carcinogenesis is associated with decreased CpG island methylation and increased mRNA and protein expression of the colonic vitamin D receptor. AB - Epidemiological studies suggest that estrogen prevents neoplastic transformation in the intestinal mucosa. Estrogen was shown to increase the expression of vitamin D receptors (VDR) in a variety of tissues. 1,25-Dihydroxyvitamin D [1,25 (OH)2D] and several of its analogues are known as potent antineoplastic and prodifferentiative in many cell types, including colon-derived cells. The present study was designed to examine the effect of estradiol (E2) on dimethylhydrazine (DMH)-induced colon cancer in rats, and the possibility that E2 may exert its protective effect on the colon through modulation of the vitamin D-endocrine system. The in vivo effect of E2 on DMH-induced colorectal cancer was studied in four groups of ovariectomized female rats: (I) untreated control, (II) E2 treated, (III) DMH treated, and (IV) combined E2 and DMH treated. Significantly higher uterine weights and higher colonic estrogen receptor content confirmed the effectiveness of ovariectomy and E2 replacement. The number of malignant tumors in group IV was 2.3+/-1.1 (mean +/- SE) per rat, compared with 8.1+/-1.9 in group III (P < 0.001). Exposure to estrogen was associated with a marked increase in VDR mRNA content and VDR protein expression in the normal colonic mucosa. In tumor extracts VDR protein expression was considerably lower compared with normal mucosa. Estrogen treatment did not affect serum levels of 25(OH)D, 1,25(OH)2D, and PTH. Significant CpG island methylation in the VDR gene was observed in colonic tissue DNA harvested from rats treated with DMH, but not in colonic mucosae from rats treated with DMH + E2. The highest frequency of CpG methylation in the VDR gene was detected in DNA extracted from cancer tissue rims. In summary, the protective effect of estrogen against chemically induced colonic carcinogenesis is associated with reduced methylation of the VDR gene and with upregulation of both VDR gene transcription and protein expression. We suggest that estrogen may interfere with the process of CpG DNA methylation in the colonic mucosa to prevent silencing of the VDR gene. Increased VDR activity could be one of the mechanisms by which estrogen protects against neoplastic transformation in the colon. PMID- 10691029 TI - Growth inhibition of a human ovarian tumor by a novel paclitaxel derivative in SCID mice. AB - We report here the toxicity and therapeutic effects of 2'-alpha-bromohexadecanoyl paclitaxel (BrC16HT), a prodrug form of paclitaxel, in mice. Paclitaxel is the active ingredient of Taxol. The maximum tolerated dose, at a one dose per day for 5-day schedule, was 37.5 mg/kg for BrC16HT compared to 12.5 for Taxol administered IP, and was 12.5-25 mg/kg for either agent administered IV. Dose dependent therapeutic effects were found for BrC16HT against a human ovarian tumor (OVCAR-3) grown in SCID mice. IP treatments with BrC16HT against early or established IP-implanted OVCAR-3 tumor increased mean survival times more than treatment with Taxol. Long-term survivors were found only in groups treated with BrC16HT. Intravenously administered BrC16HT was more effective than Taxol against SC OVCAR-3 tumor. Early treatment (25 mg/kg x 5) completely inhibited tumor growth through 120 days after tumor implantation. Pharmacokinetic studies suggest that BrC16HT is slowly hydrolyzed to paclitaxel and circulates longer than paclitaxel from Taxol. Thus, BrC16HT may provide sustained levels of paclitaxel, which may contribute to the increased efficacy of BrC16HT compared to Taxol. PMID- 10691030 TI - Identification of seven genes regulated by wild-type p53 in a colon cancer cell line carrying a well-controlled wild-type p53 expression system. AB - We applied a differential display method to screen mRNAs isolated from a newly established cell line that carried a wild-type p53 transgene under control of the lactose operon. To investigate the p53 signaling pathway, we looked for genes whose expression was significantly induced or suppressed by induction of wild type p53 protein, and identified seven. DNA sequence analyses revealed that the two genes that were upregulated encoded isozyme 6 of aldehyde dehydrogenase (ALDH6) and subunit I of cytochrome c oxidase (COI). The five genes that were downregulated encoded protein-tyrosine kinase (Syk), high mobility group chromosomal protein 17 (HMG-17), transferrin receptor, human alpha-tubulin, and sds22-like protein. The results indicated that genes related to cell cycle regulation, cell respiration, and cytoskeletal structure are involved in the process of growth arrest induced by wild-type p53. PMID- 10691031 TI - Neuroprotective interactions in rats between paclitaxel and cisplatin. AB - Paclitaxel and cisplatin are associated with dose-limiting neurotoxicity that may result from their differing effects on microtubule stability in peripheral nerves. We hypothesized that such different actions of paclitaxel and cisplatin could be exploited to minimize their neurotoxicity by giving them in combination. Paclitaxel (9-18 micromol/kg/week or 7.7-15.4 mg/kg/week) and cisplatin (5-10 micromol/kg/week or 1.5-3 mg/kg/week) were given alone and in combination to female Wistar rats. Treatment was given once per week for a total of 7-10 weeks. Paclitaxel and cisplatin were given 24 h apart when they were given in combination. Changes in sensory nerve conduction velocity (SNCV) and dorsal root ganglia (DRG) morphology were measured. The nature of their interaction was analyzed using an isobologram. Their antitumor activity alone or in combination was also determined in C57B1/6 mice bearing colon 38 tumors. Reductions in SNCV occurred with paclitaxel alone (P = 0.009), cisplatin alone (P = 0.012), and cisplatin given 24 h before paclitaxel (P < 0.0001). In contrast, there was no significant change in SNCV with paclitaxel given 24 h before cisplatin (P = 0.11). An isobologram showed that the SNCV effects of the drug combinations were less than additive or antagonistic. Cisplatin-induced morphometric changes in DRG neurons were less marked when cisplatin was given with paclitaxel (P = 0.004). Concentrations of platinum in dorsal root ganglia, sural nerves, and sciatic nerves were not altered by giving paclitaxel before cisplatin. Tumor growth delays (TGD) were greater after treatment with paclitaxel (23.4 micromol/kg or 20 mg/kg) given 24 h before cisplatin (23.3 micromol/kg or 7 mg/kg) (TGD = 7.5 days) than after paclitaxel (23.4 micromol/kg or 20 mg/kg) (TGD = 2.0 days) or cisplatin (23.3 micromol/kg or 7 mg/kg) (TGD = 3.5 days) alone. Paclitaxel and cisplatin antagonized each other's neurotoxicity in Wistar rats. Combining cytotoxic agents with opposing effects on peripheral nerves has potential for minimizing neurotoxicity in patients. PMID- 10691032 TI - Supplementation with soybean lipids reduces goat serum-induced apoptosis in the B cell hybridoma CC9C10. PMID- 10691033 TI - In situ labeling of adherent cells with PKH26. PMID- 10691034 TI - Primary cultures of midgut cells from Heliothis virescens can be frozen and stored. PMID- 10691035 TI - Activation of the epidermal platelet-activating factor receptor results in ICAM-1 expression. PMID- 10691036 TI - The effect of retinoic acid on the proportion of insulin cells in the developing chick pancreas. AB - We assessed the potential role of all-trans-retinoic acid on the developing chick pancreas, specifically with regard to the proportions of insulin cells. The endodermal component of the dorsal pancreatic bud of 5-d-old chick embryos was cultured on Matrigel. Retinoic acid (10(-6) or 10(-5) M) was added to a standard serum-free medium, Ham's F12 containing insulin, transferrin and selenium (F12.ITS). Control grafts were cultured in F12.ITS alone or in F12.ITS with DMSO (the diluent for retinoic acid). After 7 d the explants were retrieved, freeze dried, vapor-fixed, and embedded in resin. Endocrine cell types were identified by immunocytochemistry. The numbers of insulin cells were expressed as a proportion of the sum of insulin plus glucagon cells. Retinoic acid had a dose related effect; the proportion of insulin cells in explants treated with the lower dose of retinoic acid (10(-6) M) was more than twice the proportion of insulin cells in explants treated with the higher dose (10(-5) M) of retinoic acid and more than three times that of the control grafts. PMID- 10691037 TI - Partial cloning and sequencing of chick fibrillin-1 cDNA. AB - The recent identification of numerous matrix genes and gene products has allowed a detailed examination of their roles in development. Two of these extracellular matrix proteins, fibrillin-1 and fibrillin-2, are components of the elastin associated microfibrils. Given what is known about the distribution of the fibrillins in normal tissues and the abnormalities that result when mutations occur, a basic hypothesis has emerged: fibrillin-1 is primarily responsible for load bearing and providing structural integrity, whereas fibrillin-2 may be a director of elastogenesis. Nevertheless, examination of phenotypes in disorders caused by mutations in fibrillin-1 or fibrillin-2 suggests some common functions. To better understand these similar and diverse roles, it would be helpful to examine these proteins during chick development. To accomplish this goal, it is first necessary to characterize the chick homologs of the known fibrillins. In this study, the partial chick FBN1 cDNA was identified by polymerase chain reaction-aided cloning as a first step toward elucidating these goals. Sequence analysis indicated that there is striking conservation between chick and mammalian fibrillin-1 at the DNA and protein levels. Antisense and sense riboprobes were synthesized and used in in situ hybridization in stage 14 chick embryos and high levels of FBN1 transcripts were observed in the heart. PMID- 10691038 TI - Establishment and characterization of a caprine mammary epithelial cell line (CMEC). AB - We describe the establishment of a continuous, nontransformed cell line obtained from primary culture of a lactating (114 days postparturition) Anglo-Nubian (Capra hircus) goat mammary gland biopsy. These cells (CMEC), have been cultured in the presence of supraphysiologic concentrations of insulin and hydrocortisone for more than 560 population doublings (over 80 passages) without any sign of senescence while maintaining a normal/near-normal diploid chromosome modal number of 2n = 60 and are responsive to contact inhibition of proliferation. Cytoskeletal analysis indicates that CMECs are epithelial, without detectable fibroblastic or myoepithelial cells. When grown at low density on plastic substratum, the cells tend to form island monolayer aggregates with the characteristic cobblestone morphology of epithelial cells. With increasing density, the cells organize into lumen-like structures with various morphology consisting of large and small vacuolized and nonvacuolized cells. Postconfluent cultures form epithelial raised dome-like structures, implying a process of contact-induced differentiation. This is corroborated by positive immunocytochemistry to lactation-specific proteins: beta-casein and alpha lactalbumin, which were predominantly expressed in dome-forming cells. We also observed an overall modulation of cytokeratin 18/19 expression associated with number of days post subculture and with the expression of lactation-specific proteins. Postconfluent cultures which contain lactation-specific, antibody reactive, dome-like structures showed a decreased expression of keratin 18 and no (null) expression for keratin 19. Lastly, cells cultured within a collagen matrix show morphological differentiation with the organization of branching duct-like and acini-like structures. This study suggests that CMECs are a useful in vitro model for study of mammary gland development and differentiation, in particular, direct modulation of epithelial cells grown on plastic substratum or extracellular matrix without the influence of stromal elements or the necessity and variability associated with primary cell culture or tissue explants. PMID- 10691039 TI - Organization of extracellular matrix components during differentiation of adipocytes in long-term culture. AB - Scanning electron microscopy (SEM) observation showed that fully differentiated spherical adipocytes were embraced by a network of collagens and fibroblastic preadipocytes. The properties of both the collagen networks and the preadipocytes allow the adipocytes to be interconnected, forming a fat-cell cluster, which can anchor to the bottom of a culture dish. In this network structure, collagen fibrils and fibrillar bundles were closely arranged and stratified. We found that immunostained collagens appeared to form extracellular network structures, which can be observed by SEM. The extracellular network of fibronectin was the first to develop among the extracellular matrix (ECM) components, though it became degraded with the progress of adipocyte differentiation. The type I collagen network was the last to develop and remained well organized through the late stage of adipocyte differentiation. The extracellular networks of type III, V, and VI collagen developed by the mid-stage and remained in the late stage of adipocyte differentiation. The network structures of type IV collagen and laminin became degraded during the differentiation process and localized at the surface of spherical cells. In addition to these basement membrane components, types III, V, and VI collagens also showed pericellular spherical staining patterns. These results demonstrated that the constitution and distribution of the ECM are altered during adipocyte differentiation, suggesting that the organization of each ECM component into a suitable structure is a requirement for the differentiation and maintenance of unilocular adipocytes. PMID- 10691040 TI - Establishment of astrocyte cell lines from sheep genetically susceptible to scrapie. AB - Primary cultures of the brain from sheep embryos were used to establish cell lines after transfection by the simian virus 40 (SV40) large T gene. Two of the lines (A15 and 4A6) displayed astroglial properties. They expressed the glial fibrillary acidic protein (GFAP), intermediate filament protein vimentin, and S 100 (beta-subunit) protein. While numerous rodent and human glial cell lines are available, this is to our knowledge the first description of ovine cell lines with astrocyte features. In addition, these cell lines were derived from sheep embryos chosen for their genetic susceptibility to scrapie (PrP genotype: VV136, QQ171). Therefore, they could be attractive tissue culture models for the study of propagation and pathogenesis of the scrapie agent ex vivo. PMID- 10691041 TI - Simultaneous measures of contraction and intracellular calcium in single, cultured smooth muscle cells. AB - Simple methods are presented for quantitating contraction and intracellular calcium simultaneously in single, cultured smooth muscle cells. These methods are the first to demonstrate that reliable velocities of cell shortening can be measured in cultured smooth muscle cells and that cells in vitro exhibit shortening velocities comparable to those measured in the fastest phasic muscles in situ. Temporal relationships between changes in intracellular calcium and shortening within single cells were determined with a resolution of 100 ms and were consistent with measures in more "classical" preparations. Intracellular calcium rose quickly and transiently 10-fold above the basal level of 80-90 nM in response to the muscarinic agonist, carbachol. Shortening of the cells occurred 200 ms after intracellular calcium began to rise. The sensitivity and reliability of these methods allowed the effects of different stimuli to be easily resolved. The present report demonstrates that genuine contractility need not be ignored in cultured smooth muscle cells and that the temporal relations between shortening and intracellular calcium mobilization can be quantitatively assessed in controlled in vitro environments. PMID- 10691042 TI - Modulatory effect of rat small intestinal epithelial cell-conditioned medium on lymphocyte proliferation. AB - The small intestinal epithelium plays an important role in the mucosal host defense. Intestinal epithelial cells have been known to release substances that suppress lymphocyte proliferation, suggesting an immunoregulatory function. We investigated how intestinal epithelial cells affect lymphocyte proliferation. Serum-free medium that was conditioned by incubating epithelial cells, particularly crypt cells, of the rat small intestine affected proliferation of allogeneic spleen lymphocytes stimulated with concanavalin A, as assessed by measuring cellular [3H]thymidine incorporation. Less than 1% and greater than 2% of the conditioned medium enhanced and suppressed, respectively, lymphocyte proliferation. The causative substances found in the conditioned medium were dialyzable and heat-stable. Suppression was not due to toxicity to splenocytes. Exposure of splenocytes to a suppressive concentration of the conditioned medium beginning at 30 min before an onset of lectin stimulation decreased the suppression of lymphocyte proliferation. Splenocyte exposure to the suppressive concentration of the conditioned medium beginning at 30 min to 4 h after the onset of the stimulation inversely strengthened the suppression. A brief exposure of splenocytes to the conditioned medium for the last 4 h during a total 72-h culture period still suppressed lymphocyte proliferation. Thus, intestinal epithelial cells produce low-molecular-weight lymphocyte proliferation-modulating substances that suppress the proliferation of lectin-activated lymphocytes, but not resting ones, by affecting earlier intracellular events and the following DNA synthesis when incubated in culture medium. PMID- 10691043 TI - Cytotoxicity of TNT and its metabolites. AB - The production and storage of explosives has resulted in the environmental accumulation of 2,4,6-trinitrotoluene (TNT). The biotransformation products of the nitroaromatic compound TNT and metabolites in mammalian cells in culture and their cytotoxicity are studied. We report after our analysis by reverse phase high performance liquid chromatography (HPLC) that the most prevalent biotransformation product of TNT in the NG108 neuroblastoma cells is primarily monoamino-dinitrotoluene (2Am-DNT). It causes toxic effects based on trypan blue exclusion and LDH-release colorimetric assays. PMID- 10691044 TI - Diagnosis of myocardial infarction: integration of serum markers and clinical descriptors using information theory. AB - OBJECTIVE: We examine the use of information theory applied to a single cardiac troponin T (cTnT) (first generation monoclonal; Boehringer Mannheim Corp., Indianapolis, Indiana) used with the character of chest pain, electrocardiography (ECG) and serial ECG changes in the evaluation of acute myocardial infarction (AMI). We combined a single measure of cTnT (blinded to the investigators) with a creatine kinase MB isoenzyme (CK-MB) measurement to discover the best decision value for this test in a study of 293 consecutive patients presenting to the emergency department with symptoms warranting exclusion of AMI. METHODS: The decision value for determining whether cTnT is positive or negative was determined independently of the final diagnosis by examining the information in the cTnT and CKMB data. Using information theory, an autocorrelation matrix with a one-to-one pairing of the CKMB and troponin T was constructed. The effective information, also known as Kullback entropy, assigned the values for troponin T and for CKMB that have the lowest frequency of misclassification error. The Kullback entropy is determined by subtracting the data entropy from the maximum entropy of the data set in which the information has been destroyed. The assignment of the optimum decision values was made independently of the clinical diagnoses without the construction of a receiver-operator characteristic curve (ROC). The final diagnosis of AMI was independently determined by the clinicians and entered into the medical record. RESULTS: The decision value for cTnT was 0.1 ng/ml as determined by the the information in the data. The method was validated within the same study by mapping the results so obtained into the diagnoses obtained independently by the clinicians using all of the methods at their disposal. The cTnT was different in AMI (n = 60) compared with non-AMI patients (n = 233) (2.08 +/- 0.21 vs. 0.07 +/- 0.10; p < .0001). CONCLUSION: Information theory provides a strong framework and methodology for determining the decision value for cTnT which minimizes misclassification errors at 0.1 ng/ml. The result has a strong correlation with other features in detecting AMI in patients presenting with chest pain. PMID- 10691045 TI - A 70-year-old man with isolated weight loss and a pellagra-like syndrome due to celiac disease. AB - An elderly man was diagnosed with celiac disease, which presented with three notable features: first, presentation at the age of 70 with no prior gastrointestinal symptomatology or positive family history; second, triggering of all symptoms following recent myocardial infarction and infective endocarditis; third, presentation with marked (more than 20 percent) weight loss and pellagra like skin lesions despite nearly normal examination and laboratory tests. Thus, celiac disease may present as a pellagra-like syndrome in the elderly with predominant weight loss that is enhanced by the related taste disturbances. PMID- 10691046 TI - Advancing technologies in clinical medicine: the Yale-Mount Everest telemedicine project. PMID- 10691047 TI - Technical and cultural challenges of remote health care on Everest. PMID- 10691048 TI - For once then, something... PMID- 10691049 TI - Relationships among serum cystatin C, serum creatinine, lean tissue mass and glomerular filtration rate in healthy adults. AB - In an effort to increase our knowledge of the optimal use of serum cystatin C and creatinine as glomerular filtration rate (GFR) markers, these variables, as well as lean tissue mass and GFR, were determined in a population of 42 healthy young adults (men and women with normal GFR). Dual-energy X-ray absorptiometry and measurement of the plasma clearance of iohexol were used to measure lean tissue mass and GFR, respectively. Serum creatinine was significantly correlated to lean tissue mass (r=0.65; p < 0.0001) but not to GFR (1/creatinine vs. GFR: r=0.11; p=0.106). In contrast, serum cystatin C correlated with GFR (1/cystatin C vs. GFR: r=0.32; p=0.0387), especially in men (1/cystatin C vs. GFR: r=0.64; p=0.0055), but not to lean tissue mass. These results might explain previous observations that serum cystatin C seems to be a better marker for GFR than serum creatinine, particularly for individuals with small to moderate decreases in GFR. However, the results also show that the serum concentrations of both creatinine and cystatin C are determined not only by GFR, but also by other factors. Since these additional factors differ for cystatin C and creatinine, it seems justified to use serum creatinine and cystatin C in conjunction to estimate GFR, at least until it is known in what situations serum creatinine or cystatin C is the preferable marker. PMID- 10691050 TI - Importance of assay conditions in visualization and quantitation of serum alkaline phosphatase isoenzymes separated by electrophoresis. AB - The importance of separation and identification of serum alkaline phosphatase (ALP; E.C. 3.1.3.1) fractions/isoenzymes has been frequently reported. Each serum ALP fraction/isoenzyme quantitation has both practical and theoretical importance. In the present work, serum was collected from Wistar rats and, in identical experimental conditions, total serum ALP activity and serum ALP electrophoretic fractions/isoenzymes activities were quantified. Different results for both kinds of ALP activity were obtained when different buffers or mixture of these buffers (carbonate/bicarbonate; 2-amino-2-methyl-1-propanol/HCl; Veronal, sodium diethylbarbiturate/HCl), pH conditions (9.4 and 10.4) and substrates (alpha- and beta-naphthyl phosphates) were used. Higher total serum ALP activity was always observed with beta-naphthyl phosphate, independently of the buffer (or mixture of buffers) and pH used. Electrophoresis allowed the separation of two serum ALP fractions. Activity of both serum ALP electrophoretic fractions was always higher with beta-naphthyl phosphate, except with carbonate/bicarbonate pH 10.4. The effect of a change in pH was buffer- (or mixture of buffers) and substrate-dependent; the addition of a second buffer (to that previously used) was not always accompanied by an increase or decrease (of the same magnitude) in our results. The results obtained with different buffers (or mixture of buffers) were not identical with substrates and pH values. It is concluded that (i) from the same electrophoretic separation of serum ALP fractions/isoenzymes, different values for its activity can be obtained by changing the assay conditions used for ALP visualization (revelation, staining); (ii) the same assay conditions for quantitation of total serum ALP and serum ALP electrophoretic fractions/isoenzymes should be used; (iii) the choice of assay conditions should take into account the biochemical problem being studied in each case. PMID- 10691051 TI - Metabolic syndrome is associated with changes in D-mannose metabolism. AB - Serum mannose concentration increases in diabetic patients and correlates closely with blood glucose. In patients with glomerulonephritis, serum mannose and mannose/glucose ratio positively correlate with dyslipidemia and the extent of urinary protein excretion. We investigated whether changes in serum mannose mark subjects with features of metabolic syndrome, including obesity, hypertension, glucose intolerance, and dyslipidemia. The study comprised 20 patients with mean age of 68 (SD 11) years, body mass index 27.2 (SD 5.1) kg/m2, blood glucose 6.2 (SD 1.6) mmol/L, serum total cholesterol 6.3 (SD 1.2) mmol/L, triglyceride 2.0 (SD 0.08) mmol/L, uric acid 320 (SD 109) micromol/L, mannose 60.0 (SD 17) micromol/L, and mannose/glucose ratio 9.7 (SD 1.8) micromol/mmol. Serum mannose correlated with blood glucose (r=0.758, p=0.012), triglyceride (r=0.478, p=0.023), and HDL-cholesterol (r = approximately 0.427, p=0.022). Mannose/glucose ratio correlated with BMI (r=0.581, p=0.033), mannose (r=0.491, p=0.035), and uric acid (r=0.608, p=0.027). The rate of VLDL lipoprotein turnover may be instrumental in the regulation of serum mannose concentration. We conclude that an altered mannose metabolism is a novel consideration among the metabolic abnormalities in the metabolic syndrome. PMID- 10691052 TI - Variant transthyretin (TTR) amyloidosis in Argentina. Detection of the trait by electrospray ionization mass spectrometry of lyophilized TTR immunoprecipitate. AB - We have developed a quick and reliable diagnostic method for detecting variant forms of transthyretin (TTR); namely, centrifugal concentration followed by electrospray ionization mass spectrometry (ESI-MS). Argentinian patients from three families with neuropathic amyloidosis and their relatives were screened for mutated TTR by ESI-MS. In order to facilitate transportation, we investigated the impact storage had on lyophilized anti-TTR-antibody precipitates' mass spectra. For this investigation, plasma samples from three Swedish patients with known TTR amyloidosis were analysed. We detected identical, additional peaks corresponding to a variant form of TTR in 10 members of the families, and also in a lyophilized sample sent unfrozen by mail from Argentina. All except one symptomatic subject had additional peaks, the exception having undergone a liver transplantation for the disease. All patients were early onset cases, i.e. below 35 years of age, and family history suggests an aggressive, rapidly progressing disease. Lyophilized anti-TTR-antibody precipitates stored at room temperature for 1 week exhibited only minor differences compared with plasma samples stored at -70 degrees C. In a new Argentinian study on familial amyloidotic polyneuropathy, the variant TTR was quickly identified and typed by ESI-MS. To facilitate transportation, dry-frozen samples can be used and the quality of the spectra is similar to that of samples stored at -70 degrees C. PMID- 10691053 TI - Accuracy of pleural fluid pH and PCO2 measurement in a blood gas analyser. Analysis of bias and precision. AB - Pleural fluid pH measurements are used in the management of parapneumonic effusions. This study evaluated the accuracy of the pleural fluid pH and PCO2 measurement in a blood gas analyser (BGA) and the error induced by altering the aerobic environment and temperature of sample storage prior to analysis. The pH electrodes in a pH meter and a BGA were initially evaluated using aqueous reference solutions. The accuracy of pH and PCO2 electrodes was then assessed in pleural fluid tonometered to a known PCO2. The effects of aerobic compared to anaerobic storage and storage temperature, 0 degrees C vs 37 degrees C of pleural fluid were evaluated for errors in pH and PCO2 measurement. The BGA and the pH meter measured pleural pH with negligible bias and imprecision (0.01+/-0.01 pH units). The PCO2 measurement bias and imprecision of the BGA was small: 1.8+/ 0.57 torr (0.2+/-0.08 kPa) at 2% CO2, 0.1+/-0.93 torr (-0.01+/-0.1 kPa) at 5% CO2 and -3.77+/-0.81 (-0.5+/-0.1 kPa) torr at 10% CO2 level. Aerobic storage resulted in a clinically important overestimation of pleural fluid pH by both the pH meter and the BGA of 0.14-0.16 pH units (p<0.05). No significant differences were detected when storage at 0 degrees C was compared to that at 37 degrees C. A BGA is reliable for pleural fluid pH measurements. Pleural fluid PCO2 measurements in a BGA have a concentration-dependent bias and imprecision. Anaerobic storage of specimens for pleural pH and PCO2 analysis is mandatory. PMID- 10691054 TI - Hepatic and renal extraction of circulating type I procollagen aminopropeptide in patients with normal liver function and in patients with alcoholic cirrhosis. AB - The circulating level and splanchnic and renal extraction of serum type I procollagen aminoterminal propeptide (PINP) was studied in 20 patients with normal liver function and in 15 patients with alcoholic liver cirrhosis. In patients with alcoholic cirrhosis, the concentration of PINP in the femoral artery blood was significantly higher than in the group of patients with normal liver function (median 145 microg/l, 95% CI 98-195 versus 57 microg/l, 95% CI 42 92, p<0.001). A significant decrease in the concentration of PINP between the femoral artery (median 57 microg/l, 95% CI 42-92) and the hepatic vein (median 45 microg/l, 95% CI 40-70, p<0.001) was found in patients with normal liver function. In this group we also observed a significantly higher concentration of PINP in femoral artery blood (median 60 microg/l, 95% CI 45-87) as compared with that in renal vein (median 50 microg/l, 95% CI 40-65, p<0.001). In contrast, serum-PINP did not differ between arterial and hepatic or venous venous blood in patients with alcoholic cirrhosis. Size-chromatography revealed no significant change in the ratio of the high and low molecular forms of PINP following extraction in liver and kidney. It is concluded that circulating PINP is extracted in the normal liver and kidney, and that the serum concentration of PINP is significantly higher in patients with alcoholic cirrhosis than in patients with normal liver function. Both the hepatic and the renal clearance of PINP are seriously impaired/reduced in patients with alcoholic cirrhosis. PMID- 10691055 TI - Sensitive and quantitative one-step polymerase chain reaction using capillary electrophoresis and fluorescence detection for measuring cytokeratin 19 expression. AB - An improved quantitative assay to measure cytokeratin 19 (CK19) expression has been developed. The assay utilizes reverse transcription and a one-step polymerase chain reaction (PCR), with capillary electrophoresis and fluorescent labelling, to separate and detect the PCR products. Calibration curves were constructed from a serial dilution of CK19 cDNA coamplified with a fixed amount of CK19 internal standard, which was found to be linear between 10 and 500 molecules. Quantitative measurement of CK19 in samples was carried out by coamplifying the cDNA with a fixed amount of internal standard. The values were calculated from the calibration curve. The integrity of RNA and cDNA synthesis was checked by quantitative measurement of the breakpoint cluster region (BCR) gene expression. The assay is sensitive, detecting < 10 CK19 transcripts, and reproducible with a coefficient of variation of approximately 10%. CK19 expression showed overlapping values when measured in samples from peripheral blood and bone marrow in operable breast cancer patients, in healthy volunteers or patients without epithelial cancer and in blood samples from patients with metastatic breast cancer. As the assay is easier to perform than traditional quantitative competitive PCR assays, it might be useful for quantitative measurement of other specific transcripts. PMID- 10691056 TI - Rapid MEN 2A gene carrier identification using primer-specific PCR amplification. AB - DNA testing is of great importance in families with multiple endocrine neoplasia (MEN) type 2A to identify non-mutant carrying family members and asymptomatic mutation carriers, and also to confirm the diagnosis in patients who already show clinical or biochemical signs of disease. Several point mutations of the RET proto-oncogene on exons 10 and 11 are associated with the disease, which is characterized by medullary thyroid carcinoma, pheochromocytoma and hyperparathyroidism. The aim of the present study was to develop and evaluate a simple method, which indicates the mutational status of members of families where the site of the point mutation is known. The method is illustrated by the detection of mutation TGC-->TAC of codon 611, which is one of the well-known mutations associated with MEN 2A. The method involves the PCR technique with allele-specific primers and detection of the amplified sequences with biotinylated probes. There was a clear-cut difference between the readings from affected and unaffected subjects. The subjects had been evaluated independently and all subjects harboring the mutation also had clinical disease. The method provides a simple and reliable diagnostic tool for DNA screening of members of families with a known mutation of the RET-gene. PMID- 10691057 TI - Computerized measurement of LDL particle size in human serum. Reproducibility studies and evaluation of LDL particle size in relation to metabolic variables and the occurrence of atherosclerosis. AB - OBJECTIVES: The main aims of the present research project were to develop and evaluate a new software program for evaluation of LDL particle size applied to the gradient gel electrophoresis methodology without the use of previous ultracentrifugation, and to investigate the relationships among LDL particle size, metabolic variables and atherosclerosis, as measured by ultrasound, in subjects with different degrees of insulin resistance. METHODS: LDL particle size was determined by polyacrylamide gradient gel electrophoresis. RESULTS: Coefficient of variation for between-assay experiments was 0.3% (r = 0.99) for measurement of LDL peak particle size. LDL peak particle size was negatively correlated to serum triglycerides, apolipoprotein B, fasting insulin, BMI and diastolic blood pressure and positively correlated to HDL. Furthermore, subjects with moderate to large plaques in the carotid artery had smaller LDL particles compared to subjects without plaques. CONCLUSIONS: This project resulted in a highly reproducible, computerized method for the analysis of LDL particle size. The data suggest that it is possible to assess LDL particle size in serum without the use of previous ultra-centrifugation. LDL particle size was associated with metabolic variables and the occurrence of moderate to large plaques in the carotid artery. PMID- 10691058 TI - Improved method for non-denaturing polyacrylamide gradient gel electrophoresis for detection of small-sized LDL produced during postprandial hypertriglyceridaemia. AB - A reliable method for detection of small-sized low-density lipoprotein (LDL) produced during transient hypertriglyceridaemia induced by ingestion of fat is described. Electrophoresis using a commercial non-denaturing 2.0-16% polyacrylamide gradient gel is commonly utilized to determine the size of LDL particles, but it failed to detect a minor change in LDL size induced during postprandial hypertriglyceridaemia. Detection of small-sized LDL was achieved by adjusting the polyacrylamide concentration. Electrophoresis using a non denaturing 1.5-10% polyacrylamide gradient gel gave the best resolution for detecting small-sized LDL induced during postprandial hypertriglyceridaemia. This improved method may be useful for elucidating the underlying mechanism of production of small-sized LDL (small dense LDL) in subjects with chronic hypertriglyceridaemia. PMID- 10691059 TI - The disease profile of Texas prison inmates. PMID- 10691060 TI - The disease profile of Texas prison inmates. AB - PURPOSE: Whereas prison inmates are reported to exhibit poorer overall health status and higher rates of health care utilization than the general population, no current information exists on the overall disease profile of the U.S. prison population. The present study examined the prevalence of major acute and chronic conditions in one of the nation's largest prison populations. METHODS: The study population consisted of 170,215 Texas Department of Criminal Justice (TDCJ) inmates who were incarcerated between August 1997 and July 1998. Information on medical conditions and sociodemographic factors was obtained from an institution wide medical information system. RESULTS: Infectious diseases (29.6%) constituted the most prevalent major disease category among inmates. This was followed by diseases of the musculoskeletal system and connective tissue (15.3%), diseases of the circulatory system (14.0%), mental disorders (10.8%), and diseases of the respiratory system (6.3%). Among the specific conditions examined, evidence of tuberculosis infection without active pulmonary disease (20.1%) was found to be the most prevalent condition, followed by hypertension (9.8%), asthma (5.2%), low back pain (5.1%), and viral hepatitis (5.0%). CONCLUSIONS: The present study shows that for a number of conditions, the prison population exhibited prevalence rates that were substantially higher than those reported for the general population. Moreover, estimates for a number of diseases varied substantially according to age, race, and gender. Understanding the disease profile in U.S. incarcerated populations will permit correctional administrators to develop more efficient health care delivery systems for prison inmates. PMID- 10691061 TI - Application of computer tomography-oriented criteria for stroke subtype classification in a prospective study. AB - PURPOSE: To apply stroke subclassification criteria based on computer tomography (CT) to strokes in the Nurses' Health Study and to assess reliability and validity of the criteria. METHODS: Among 121,701 women aged 30-55 years at entry, subclassification criteria were applied to 1369 incident strokes which occurred from 1976 to 1994. Reproducibility of the subclassification criteria was assessed in a systematic sample of 100 strokes reviewed by two independent reviewers. As a validation, relative risks (RR) for stroke subtypes were examined in a prospective analysis of 112,282 women aged 34 to 59 years, free of cardiovascular disease and cancer in 1980, with follow-up through 1994. RESULTS: Strokes were subclassified as follows: 226 subarachnoid hemorrhages, 135 intraparenchymal hemorrhages, 103 embolic, 217 large-artery occlusive, 309 lacunar, and 97 other thrombotic infarctions, as well as 282 strokes of undetermined type. No intercoder discrepancies were found in classification of subarachnoid hemorrhage, intraparenchymal hemorrhage, or embolic infarction, whereas there were four discrepancies relative to subclassification of thrombotic infarction, mostly due to inconsistent documentation of CT findings. In multivariate models, associations of older age (> 60 years), current smoking, history of diabetes, and high cholesterol with stroke subtypes were consistent with previous epidemiologic, clinical, or pathologic findings. CONCLUSIONS: Stroke diagnostic criteria based primarily on CT were found to have a high rate of reliability and validity. These stroke subclassification criteria may be useful in examining whether associations for various lifestyle and health behaviors differ with stroke subtypes. PMID- 10691062 TI - Measuring the projected public health impact of lung cancer through lifetime and age-conditional risk estimates. AB - PURPOSE: Lifetime and age-conditional probability (risk) estimates of developing lung cancer in the United States are presented by age, race, and gender. Effects on the risk estimates of an aging population and changing tobacco use are identified. METHODS: Risk estimates were derived by applying cross-sectional, population-based incidence rates of malignant lung cancer and mortality rates from other causes to a hypothetical cohort. The cohort was aged through a double decrement life table to determine the expected proportion of the population that would develop the disease across age intervals. Incidence and mortality data were obtained from the Surveillance, Epidemiology, and End Results (SEER) Program and the National Centers for Health Statistics, respectively. RESULTS: Among all cancers, risk estimates of developing lung cancer within 10 years, conditioned on being free of the disease at age 50, 60, or 70, ranked second to prostate cancer for men and second to breast cancer for women. For men, despite higher incidence rates of lung cancer for blacks than whites across most age groups, the risk of developing this disease over the life-span becomes similar, because white men are more likely to live to older ages where lung cancer is common. For women, lung cancer incidence rates are similar between Whites and Blacks, but an older age distribution among white women explains their greater lifetime risk of being diagnosed with the disease. Changes in the age distribution between the mid 1970s and the mid 1990s had little impact on the short-term risk estimates of developing lung cancer for younger ages but had a large influence on long-term risk estimates, particularly for the older age groups. CONCLUSIONS: Declining lung cancer among younger age groups may be attributed to declining tobacco use among the cohorts, but several more years may be required before the trends begin to fall in older age groups, particularly in women. In the meantime, an aging population is contributing to more people being diagnosed with lung cancer. Consequently, the projected risk of developing lung cancer will remain high for several years to come. PMID- 10691063 TI - Endometrial cancer in Olmsted County, MN: trends in incidence, risk factors and survival. AB - PURPOSE: We updated an earlier study in this community from 1945-1974 in order to assess trends in the incidence of, risk factors for, and survival from endometrial cancer in 1975-1991. METHODS: Incidence rates were based on all new cases of endometrial cancer diagnosed among Olmsted County, Minnesota, women during the years 1975-1991, with the population denominator from decennial census data. Risk factors were assessed with conditional logistic regression comparing the incidence cases to age- and gender-matched controls with intact uteri seen the same year the case was diagnosed. Survival was assessed using the Kaplan Meier method. RESULTS: The incidence of endometrial cancer (age-adjusted to 1970 United States total) in 1975-1991 was 14.3 per 100,000 person-years, which is slightly increased from 1965-74. The rate was 21.7 per 100,000 person-years after adjustment for hysterectomy prevalence. As in the previous study, conjugated estrogen use for six months or more (odds ratio [OR] 2.71; 95% confidence interval [CI] 1.14-6.46) and body mass index (OR 1.06; 95% CI 1.01-1.11) increased the risk of endometrial cancer. The five-year relative survival rate (82%) was not improved over the earlier study. CONCLUSIONS: A small increase in endometrial cancer incidence was linked to the same risk factors identified in an earlier study in this community. No improvement in survival was seen. PMID- 10691064 TI - Serum vitamins, carotenoids, and angina pectoris: findings from the National Health and Nutrition Examination Survey III. AB - PURPOSE: Whether various vitamins and carotenoids can protect against ischemic heart disease remains an unsettled question. METHODS: We performed a cross sectional analysis of data from National Health and Nutrition Examination Survey III (1988-1994) and examined the associations between serum vitamins A, C, E, and B12, serum folate, red blood cell folate, serum carotenoids, and angina pectoris in a representative population-based sample of 11,327 men and women aged 35->90 years. RESULTS: After adjusting for age, sex, race or ethnicity, education, smoking status, systolic blood pressure, serum cholesterol, high-density lipoprotein cholesterol, history of diabetes mellitus, body mass index, and physical activity with multiple logistic regression analysis, no significant associations were present for any of the serum vitamin concentrations and angina pectoris. Significant linear trends were observed for serum concentrations of alpha-carotene (p < 0.001), beta-carotene (p = 0.026), and beta-cryptoxanthin (p = 0.003). Compared with participants with carotenoid concentrations in the lowest quartile, participants with concentrations in the highest quartile had odds ratios for angina pectoris of 0.45 (95% confidence interval (CI) 0.31-0.65), 0.57 (95% CI 0.38-0.86), and 0.57 (95% CI 0.38-0.84) for alpha-carotene, beta carotene, and beta-cryptoxanthin, respectively. CONCLUSIONS: These results provide little support for a cross-sectional association between angina pectoris and serum and red blood cell folate concentrations or concentrations of vitamins A, C, E, and B12. Several serum carotenoid concentrations were associated with a reduced risk for angina pectoris, however. PMID- 10691065 TI - Effects on subject response of information brochures and small cash incentives in a mail-based case-control study. AB - PURPOSE: To investigate the impact on subject response of an information brochure and cash incentives included with mailed questionnaires in case-control studies. METHODS: A randomized trial was carried out within a case-control study investigating cancer in the Province of Ontario. Brochures were included with half of the mailed questionnaires sent to 7487 cases and 2561 controls. Controls were also sent cash incentives of $2, $5, or no money. RESULTS: With the brochure, response changed from 75.0% to 75.8% in cases, and from 70.3% to 71.1% in controls. Adjusting for differences in age, residence, sex, and cancer site/status, the change was 0.2% [95% confidence interval (CI) = -1.7-2.1] in cases, and 0.6% (95% CI = -3.1-4.3) in controls. The $2 and $5 incentives increased overall response in controls from 61.9% to 72.8% and 77.2%, respectively, i.e., by 10.9% (95% CI = 6.1-15.6) and 15.1% (95% CI = 10.4-19.7), after adjustment. This effect was largely confined to urban areas (for $2 and $5, respectively: 5.5% and 14.2% in Toronto, 15.3% and 20.4% in other urban areas vs. 2.7% and 1.0% in rural areas; p = 0.02). Response time showed little or no improvement when the brochure was included, but was markedly reduced for both the $2 and $5 incentives. CONCLUSIONS: Cash incentives can improve subject response in epidemiologic studies, whereas information brochures do not appear to have an effect. PMID- 10691067 TI - Bier block exsanguination: a volumetric comparison and venous pressure study. AB - OBJECTIVES: Intravenous regional anesthesia (IVRA) is a useful ED anesthetic technique. However, venous pressure elevation during injection can cause anesthetic leakage and toxicity. This is minimized by preinjection limb exsanguination. Although standard, Esmarch exsanguination is intolerable with limb trauma. Thus, the authors' objective was to study alternative methods. METHODS: Volunteers had upper limb exsanguination performed by Esmarch bandage, arm elevation/arterial compression (AE/AC), and a pneumatic vinyl splint. Resultant volume changes, measured by volume displacement, were normalized, and expressed as percent decreases from baseline. Volume changes of all three methods were compared. The physiologic effectiveness of the AE/AC method was tested by measuring IV pressures during simulated IVRA. Attainment of maximum venous pressure (MVP) indicated leakage under the tourniquet. RESULTS: All methods reduced limb volume compared with baseline (p < 0.05). No difference occurred between AE/AC and vinyl splint exsanguination (p > 0.99), but neither method was as effective as Esmarch (p < 0.05). Gender differences were noted in absolute volumes exsanguinated, but there was no difference in percent exsanguination. The AE/AC method was the simplest procedure to perform. Peak IV pressure during simulated IVRA after AE/AC was 85 mm Hg (males), and 199 mm Hg (females) (p < 0.05). The MVP was not reached. CONCLUSIONS: While Esmarch was the most effective exsanguinating method, the two alternatives provided significant and equivalent decreases in limb volume. The AE/AC technique was physiologically effective in preventing attainment of MVP. Further studies are indicated to determine the clinical effectiveness of this technique in providing anesthesia for patients with limb trauma. PMID- 10691066 TI - Design of Physicians' Health Study II--a randomized trial of beta-carotene, vitamins E and C, and multivitamins, in prevention of cancer, cardiovascular disease, and eye disease, and review of results of completed trials. AB - PURPOSE: To assess the balance of benefits and risks of supplementation with beta carotene, vitamin E, vitamin C, and multivitamins on cancer, cardiovascular (CVD), and eye diseases. DESIGN: Physicians' Health Study II (PHS II) is a randomized, double-blind, placebo-controlled trial enrolling 15,000 willing and eligible physicians aged 55 years and older. PHS II will utilize a 2 x 2 x 2 x 2 factorial design to test alternate day beta-carotene, alternate day vitamin E, daily vitamin C, and a daily multivitamin, in the prevention of total and prostate cancer, CVD, and the age-related eye diseases, cataract and macular degeneration. PRIOR RESULTS: The final results of the recently completed Physicians' Health Study I (PHS I), a randomized, double-blind, placebo controlled trial in 22,071 healthy US male physicians, indicated that beta carotene supplementation (50 mg on alternate days) had no significant benefit or harm on cancer or CVD during more than 12 years of treatment and follow-up. In regards to cancer, there were possible benefits on total and prostate cancer in those with low baseline levels assigned to beta-carotene, a finding compatible with the Chinese Cancer Prevention Study for combined treatment with beta carotene, vitamin E, and selenium in a poorly nourished population. Further, with respect to CVD, there were apparent benefits of beta-carotene supplementation on subsequent vascular events among a small subgroup of 333 men with prior angina or revascularization. The currently available data from randomized trials of primary prevention are sparse and inconsistent for vitamin E and non-existent for vitamin C and multivitamins. For eye diseases, namely cataract and age-related macular degeneration, there are no completed large-scale randomized trials of antioxidant vitamins. CONCLUSIONS: PHS II is unique in several respects. PHS II is the only primary prevention trial in apparently healthy men testing the balance of benefits and risks of vitamin E on cancer and CVD. In addition, PHS II is the only primary prevention trial in apparently healthy men to test the balance of benefits and risks of vitamin C, multivitamins, as well as any single antioxidant vitamin, alone and in combination, on cancer, CVD, and eye diseases. Finally, PHS II is the only trial testing a priori the hypotheses that beta-carotene and vitamin E may reduce the risks of prostate cancer. Thus, PHS II will add unique as well as importantly relevant and complementary information to the totality of evidence from other completed and ongoing large-scale randomized trials on the balance of benefits and risks of beta-carotene, vitamin E, vitamin C, and multivitamins alone and in combination on prevention of cancer, CVD and eye diseases. PMID- 10691068 TI - The effects of epidermal debridement of partial-thickness burns on infection and reepithelialization in swine. AB - OBJECTIVE: Early postburn debridement of burn blisters is controversial. This study was conducted to compare rates of infection and reepithelialization in debrided vs nondebrided second-degree burns in swine. METHODS: This was a prospective, blinded, controlled, experimental trial using isoflurane anesthetized swine. Standardized partial-thickness burns were inflicted by applying an aluminum bar preheated to 80 degrees C to the backs and flanks of two young pigs for 20 seconds. In half of the burns the necrotic epidermis was manually debrided. All burns were randomly treated with octylcyanoacrylate spray (OCA) or dry gauze (C). Full-thickness biopsies were taken at 7, 10, and 14 days for blinded histopathologic evaluation. The primary outcomes were the proportions of infected burns at days 7 and 10 and the proportion of completely reepithelialized burns at day 14. Burns were considered infected in the presence of intradermal neutrophils containing bacteria (intraobserver agreement, K = 1.00). A secondary outcome was the proportion of burns with the presence of scar tissue (abnormal collagen under polarized light; intraobserver correlation, K = 0.93). Chi-square tests were used for group comparisons. This study had 90% power to detect a 40-percentage-point difference in infection rates (alpha = 0.05). RESULTS: A total of 126 biopsies from 42 burns were available for review. Infection rates were higher in the debrided burns both at day 7 (55% vs 4.5%, p < 0.001) and at day 10 (65% vs 9%, p < 0.001) after injury. The proportion of nondebrided burns that were completely reepithelialized was higher at days 10 (68% vs 0%, p < 0.001) and 14 (100% vs 65%, p = 0.003). The presence of scar tissue was more common in debrided burns (75% vs 4.5%, p < 0.001). Burns treated with OCA had fewer infections than controls (4% vs 55%, p < 0.001). Fewer OCA treated debrided burns were reepithelialized at 14 days than those that were not debrided (30% vs 100%, p = 0.001). CONCLUSIONS: Under the current study conditions, early postburn epidermal debridement of second-degree burns resulted in more infections and slower reepithelialization rates in swine. The effects of early postburn epidermal debridement in humans should be explored. PMID- 10691069 TI - Lower-extremity Doppler for deep venous thrombosis--can emergency physicians be accurate and fast? AB - Clinical diagnosis of lower-extremity (LE) deep venous thrombosis (DVT) requires confirmation by an imaging study before committing the patient to anticoagulation therapy. Studies have shown that demonstrating compressibility of leg veins under ultrasound is accurate for ruling out DVTs when performed by vascular specialists. Although LE Doppler has become the preferred test for diagnosing DVTs, it is not always available 24 hours per day. OBJECTIVES: To evaluate the accuracy and speed with which emergency physicians (EPs) could perform LE color duplex ultrasonography for the detection of DVT. METHODS: Patients presenting to an urban community emergency department (ED) between August 1, 1998, and March 3, 1999, were enrolled into this prospective study. The EPs, who underwent brief and standardized training, scanned patients at high risk for DVT with leg pain, swelling, or both. Physicians performed color duplex ultrasound examinations with compression at the common femoral and popliteal veins. The time until completion of the ED scan was recorded with a standardized method. The vascular laboratory performed a complete duplex ultrasound examination within eight hours. RESULTS: One hundred twelve patients were enrolled in the study, with 34 positive for DVT. The median examination time was 3 minutes 28 seconds (95% CI = 2 min 45 sec to 4 min 2 sec; IQR 3 min 9 sec). Times ranged from 1:02 to 18:20 minutes. The ED results had a high correlation with vascular laboratory studies, giving a kappa of 0.9 and a 98% agreement (95% CI = 95.4% to 100%). CONCLUSION: Emergency physicians can perform LE duplex ultrasound examinations accurately and quickly. PMID- 10691070 TI - Out-of-hospital intravenous cannulation: the perspective of patients treated by London Ambulance Service paramedics. AB - OBJECTIVES: Previous research has highlighted concern about infection rates in field-placed intravenous (IV) cannulae. In a study of IV placement by London Ambulance Service (LAS) paramedics, 17% of placements were judged to be inappropriate. Large variations in rates of IV placement between LAS paramedics were found. The authors' hypothesis was that placement of an IV carries disadvantages-pain, discomfort, distress, and infection-which may be unacceptable to patients. METHODS: This was a survey of all patients having an IV placed by LAS paramedics and transported to one of three London emergency departments (EDs) over a three-week period in December 1996. Patients were excluded if they had a self-inflicted injury/illness, were less than 14 years old, had no known address, or were visitors to the UK, or if their family doctor suggested it was not appropriate to contact the patient. Pain, discomfort, and distress; infection; satisfaction; understanding of the reason for cannulation; and out-of-hospital cannula use were all ascertained and analyzed with chi-square analysis. RESULTS: Thirty-nine percent of the respondents experienced some discomfort, 39% some pain, and 17% some distress. No patient reported an infection. Distress was more likely to be reported if there was no understanding of why the IV cannula was placed (chi2 [1] 6.1; p < 0.05). Further unstructured information revealed satisfaction with the IV cannulation and with general care. CONCLUSIONS: Despite the disadvantages of IV placement being reported by some respondents, overall levels of satisfaction were high, suggesting that these disadvantages were not unacceptable to patients. However, in the context of the 24,000 patients cannulated each year by LAS paramedics, "costs" to the patient are considerable. PMID- 10691071 TI - Traumatic brain injuries evaluated in U.S. emergency departments, 1992-1994. AB - OBJECTIVE: To describe the incidence and patient characteristics of traumatic brain injuries (TBIs) treated in U.S. emergency departments (EDs). METHODS: A secondary analysis was performed on data from the National Hospital Ambulatory Medical Care Survey administered from 1992 to 1994. An ED visit was determined to represent a case of TBI if the case record contained ICD-9-CM codes of 800.0 801.9, 803.0-804.9, or 850.0-854.1. RESULTS: The average annual estimate of new TBI treated in U.S. EDs was 1,144,807, equaling 444 per 100,000 persons (95% CI = 390 to 498), which represents approximately 3,136 new cases of TBI per day and accounts for 1.3% of all ED visits. Males were 1.6 times as likely as females to suffer TBI until the age of 65 years, when the female rate exceeded the male. The rate for blacks was 35% higher than that for whites. The highest overall incidence rate of TBI occurred in the less-than-5-year age group (1,091 per 100,000), closely followed by the more-than-85-year age group (1,026 per 100,000). Falls represented the most common mechanism of TBI injury, followed by motor vehicle-related trauma. CONCLUSIONS: This study underscores the ongoing need for effective surveillance of all types of TBI and evaluation of prevention strategies targeting high-risk individuals. It serves as a clinically grounded and ED-based corroboration of prior survey research, providing a basis for comparison of incidence rates over time and a tool with which to measure the efficacy of future interventions. PMID- 10691072 TI - Pantomography vs mandibular series for the detection of mandibular fractures. AB - OBJECTIVE: The two primary radiographic techniques used for the evaluation of mandible injury are a pantomographic series (PS) and the standard four-view mandibular series (MS). Despite a tenuous foundation, there is apparent bias in favor of PS compared with MS. Many emergency departments do not have ready access to the specialized equipment necessary to perform a pantomographic study. The hypothesis of this study was that a high-quality standard MS is as sensitive and specific as a PS in the detection of mandibular fractures. METHODS: This was a prospective, blinded study of 54 patients presenting with acute mandibular injury comparing MS and PS. The study design used two board-certified emergency physicians and a single staff radiologist who read a series of MS and PS films in a randomized fashion without access to clinical information or identifying patient data. The absolute number of fractures present was determined by a neuroradiologist with access to both MS and PS simultaneously as well as pertinent clinical information. RESULTS: Thirty patients had 47 mandibular fractures. The sensitivity for fracture detection for each physician was 0.85, 0.77, and 0.89 with MS and 0.79, 0.74, and 0.83 with PS (p > or = 0.51, p > or = 1.00, and p > or = 0.51, respectively, McNemar's binomial test). The specificity for fracture detection for each physician was 0.88, 0.92, and 0.96 for MS and 0.96, 1.00, and 0.92 for PS (p > 0.625, p > 0.50, and p = 1.00, respectively, McNemar's binomial test). CONCLUSIONS: A standard mandibular series is as sensitive and specific as pantomography in the detection of mandibular fractures. PMID- 10691074 TI - Primary closure of mammalian bites. AB - OBJECTIVE: Suturing of bite wounds remains controversial. The authors evaluated the incidence of wound infection in 145 mammalian bite wounds treated with primary closure. METHODS: Consecutive patients with bite wounds receiving primary closure at a university hospital ED had structured closed-question data sheets completed at the time of wound management and suture removal. Infection was determined at the time of suture removal using a previously validated definition. Data included demographics; medical history; time from injury to evaluation; wound characteristics and location; details of wound cleansing methods, debridement, foreign body removal, and wound closure methods; use of antibiotics; and follow-up wound evaluation. Proportions and 95% confidence intervals were calculated. RESULTS: One hundred forty-five mammalian bite patients were enrolled: 88 dog, 45 cat, and 12 human bites. Patients had a mean (+/-SD) age of 21 +/- 20 years; 58% were male; 86% were white; and they presented a mean (+/-SD) of 1.8 +/- 1.2 hours after injury. Bites occurred on the head and neck (57%), upper extremity (36%), and lower extremity (6%). Wounds had a mean length and width of 2.5 cm and 4.8 mm, respectively. Twelve percent involved structures deep to subcutaneous tissue. After primary wound closure, wound infections occurred in eight patients (5.5%; 95% confidence interval = 1.8% to 9.2%). CONCLUSIONS: The data suggest that carefully selected mammalian bite wounds can be sutured with approximately a 6% rate of infection. This infection rate may be acceptable in lacerations where cosmesis is a primary concern. PMID- 10691073 TI - Evaluation of the physician's ability to recognize the presence or absence of anemia, fever, and jaundice. AB - OBJECTIVE: The evaluation of the patient through a comprehensive history and physical examination is considered the cornerstone of medical diagnosis, but many studies suggest that physicians have inadequate physical examination skills. It is unknown whether these skills are reliable and whether they can be adequately acquired through training. The objective of this study was to evaluate the ability of the clinician to detect the presence and discriminate the extent of clinical anemia, fever, and jaundice in an ED or hospitalized patient. METHODS: This was a prospective observational study of a convenience sample of patients presenting to the ED or admitted to the hospital who had a rectal temperature measurement within 30 minutes prior to the observation, serum hematocrit measurement on the day of observation, or serum bilirubin measurement one day prior to the day of observation. Observers' (emergency medicine attending physicians', resident physicians', and rotating medical students') estimated serum hematocrit, rectal temperature, and serum bilirubin values were obtained after each observation. Sensitivity, specificity, positive predictive value, negative predictive value, and mean absolute difference between actual and estimated values were calculated for each observer. RESULTS: The physicians detected the presence or absence of anemia, fever, and jaundice in patients with sensitivities and specificities of approximately 70%. Their predictions varied from the measured value on average by 6.0 +/- 4.6% for serum hematocrit, 1.3 + 1.1 degrees F for rectal temperature, and 3.4 +/- 5.3 mg/dL for serum bilirubin. Observer accuracy decreased when evaluating patients with high and low measured values. CONCLUSIONS: The ability to correctly perform and interpret the physical examination appears to be independent of the observer level of training, patient ethnicity, or patient gender. The examination for pallor, warmth, and jaundice is unreliable in predicting the corresponding laboratory or electronic measurement. Certain anemic, febrile, or jaundiced patients may not be reliably detected solely by a focused physical examination. PMID- 10691075 TI - The focused abdominal sonography for trauma (FAST) examination: considerations and recommendations for training physicians in the use of a new clinical tool. AB - Focused abdominal sonography for trauma (FAST) is being used by growing numbers of emergency physicians and surgeons because it has proven to be an accurate, rapid, and repeatable bedside test for evaluating abdominal trauma victims. Controversy exists about the optimal means of FAST education and the number of examinations necessary to demonstrate competency. Most FAST educators agree that FAST education should consist of three phases: didactic, practical, and experiential. This article summarizes options and preliminary recommendations suitable for developing a FAST curriculum. PMID- 10691076 TI - Clinicopathological conference: multisystem failure in a child. PMID- 10691077 TI - Do emergency medical services dispatch nature and severity codes agree with paramedic field findings? AB - Emergency medical services (EMS) systems increasingly seek to triage patients to alternative EMS resources. Emergency medical services dispatchers may be asked to perform this triage. New protocols may be necessary. Alternatively, existing protocols may be sufficient for this task. For an existing dispatch protocol to be sufficient, it at least must accurately categorize patient condition and severity based on an external standard. OBJECTIVE: To examine the extent to which nature codes (NCs), or patient condition codes, and severity codes (SCs) currently assigned in one urban 911 center agree with paramedic field findings. The null hypothesis was that there is no routine agreement (75%) between dispatcher-assigned NC or SC and paramedic-assigned NC or SC for the same patient using the same protocol. METHODS: Emergency medical services dispatch nature and severity code data and matching out-of-hospital data were prospectively gathered over six months. Dispatch data included the NC: caller-identified problem, and the SC: dispatcher-assessed severity. Each NC is modified by one of three SCs (1, 3, or 9): 1 is emergent, 3 is urgent, and 9 is neither. Paramedics verified and/or corrected dispatcher-assigned NCs and SCs using the same dispatch protocol. RESULTS: One thousand forty usable cases fell into 33 unique NC/SC combinations. The designation of SC 1 was assigned 275 times, SC 3 was assigned 736 times, and SC 9 was assigned 24 times. The SC was missing five times. The overall NC agreement was 0.70 (95% CI = 0.697 to 0.703). The overall SC agreement was 0.65 (95% CI = 0.645 to 0.655). The NC agreement exceeded 75% for ten (59%) NC/SC combinations. The SC agreement exceeded 75% for five (29%) NC/SC combinations. There was both NC and SC agreement for four (24%) combinations: urgent breathing problems, urgent diabetic problems, urgent falls, and urgent overdoses. The greatest NC/SC disagreement occurred within emergent and urgent traffic crashes. Paramedics adjusted SC toward lower severity 29% of the time and toward higher severity 5.4% of the time. There was no upward SC adjustment for eight (47%) combinations. CONCLUSIONS: Certain dispatcher-assigned NC and SC codes and NC/SC combinations achieved the study threshold. Overall agreement failed to achieve the threshold. The lowest SC level was rarely assigned, preventing a meaningful analysis of all severity levels. PMID- 10691078 TI - Rattlesnake venom-induced thrombocytopenia response to Antivenin (Crotalidae) Polyvalent: a case series. AB - OBJECTIVE: To test the hypothesis that rattlesnake venom-induced thrombocytopenia would improve following Antivenin (Crotalidae) Polyvalent administration, and that the degree of platelet increase would correlate with the dosage of antivenom. METHODS: The authors conducted a retrospective review of all patients admitted for rattlesnake envenomation at two southern California hospitals between 1980 and 1998. Patients were included if platelet count was less than 150 x 10(9)/L following a rattlesnake bite. Patients were excluded if they received platelet transfusion. The relationship between Antivenin (Crotalidae) Polyvalent administration and venom-induced thrombocytopenia was evaluated by linear regression and paired t-test. RESULTS: The authors identified 103 cases of rattlesnake envenomation. Seventeen cases met inclusion criteria for thrombocytopenia. Two patients were excluded because they received platelet transfusions. One additional patient was excluded from paired t-test only because no antivenom was given. Thrombocytopenia usually improved between presentation and discharge (mean difference, 44 x 10(9)/L), although complete resolution was often not achieved. A statistically significant partial improvement in platelet counts immediately after antivenom administration was observed in a subset of patients with severe thrombocytopenia (platelet count <100 x 10(9)/L) (mean difference, 64 x 10(9)/L). Using regression analysis, the authors did not detect a linear relationship between the amount of Antivenin (Crotalidae) Polyvalent administered and the degree of improvement. CONCLUSIONS: Although rattlesnake venom-induced thrombocytopenia usually improves immediately after Antivenin (Crotalidae) Polyvalent administration and by the time of discharge, the degree of improvement is frequently incomplete and of uncertain clinical significance in the absence of life-threatening bleeding. The authors found no correlation between the degree of improvement and the dosage of Antivenin (Crotalidae) Polyvalent. PMID- 10691079 TI - Whose turf is it, anyway? Diagnostic ultrasonography in the emergency department. PMID- 10691080 TI - Outcomes research and emergency medical services: the time has come. PMID- 10691081 TI - The clinicopathological conference. PMID- 10691082 TI - Research fundamentals: statistical considerations in research design: a simple person's approach. AB - A basic understanding of statistical methodology is essential, both for designing quality research projects and for evaluating the medical literature. Careful statistical planning, including the selection of study endpoints, the determination of the required sample size, and the selection of statistical tests to be used in the data analysis, is important to ensure a successful research project. The purpose of this article is to provide a basic review of statistical terms and methods for both the researcher and the clinician, as well as to clarify questions that need to be answered prior to embarking on an experimental study. The advantages of collaborating with statistical consultants, and some guidelines for such collaborations, are discussed as well. PMID- 10691083 TI - Gestational outcome in patients with first-trimester pregnancy complications and ultrasound-confirmed live intrauterine pregnancy. PMID- 10691084 TI - A truly emergent problem: button battery in the nose. PMID- 10691085 TI - Radial head subluxation. PMID- 10691086 TI - Resident self-scheduling: a painful method of reinventing the wheel. PMID- 10691087 TI - Pneumocystis carinii infection in young non-immunosuppressed rabbits. Kinetics of infection and of the primary specific immune response. AB - The aim of this study was to determine the kinetics, the dissemination of the infection and the immunological response to Pneumocystis carinii primary infection in a non-immunosuppressed rabbit model. For this purpose, we developed a nested PCR that amplified a portion of the mitochondrial large-subunit rRNA gene of rabbit-derived P. carinii. The PCR detected P. carinii DNA in lung and bronchoalveolar lavage fluids from 14- to 45-day-old rabbits but not in their serum. No P. carinii DNA was detected in extrapulmonary organs from 28-day-old rabbits with P. carinii pneumonia. ELISA and immunoblotting analysis showed that 5-day-old pups had elevated specific IgG. The IgG concentration sharply decreased, reaching a trough on day 21, and from then onwards progressively increased as the infection cleared. Conversely, the specific IgM concentration increased during the infection and peaked on day 28. IgG mainly recognized a 50 kDa subunit of P. carinii organisms; IgM recognized first a 45-kDa subunit on day 21, whereas from day 28 onwards it also recognized the 50-kDa subunit. A P. carinii-specific splenocyte proliferative response was observed on day 45. These findings suggest that P. carinii primary infection is a time-limited and a lung limited event and contribute new information on the relationship between the kinetics of primary P. carinii infection and the immunological response in a model that mimics the primary infections in humans. PMID- 10691088 TI - The role of tumor necrosis factor (TNF)-alpha in the antitumor effect of intrapleural injection of Lactobacillus casei strain Shirota in mice. AB - The involvement of several cytokines in the antitumor effect induced by intrapleural (i.pl.) injection of heat-killed cells of Lactobacillus casei strain Shirota (LC 9018) in mice was investigated. Injection of LC 9018 i.pl. into Meth A fibrosarcoma (Meth A)-bearing mice not only significantly prolonged the survival of the mice, but also effectively inhibited the accumulation of malignant pleural fluid in the thoracic cavity. In the thoracic cavity of tumor bearing mice treated with LC 9018, we observed large amounts of several cytokines including interleukin (IL)-1beta, interferon (IFN)-gamma, IL-12 and tumor necrosis factor (TNF)-alpha. Both anti-IFN-gamma and anti-IL-12 monoclonal antibody (mAb) treatments partially diminished the antitumor activity of LC 9018 in vivo, while the treatment of anti-IL-1beta mAb did not influence the survival of the mice. However, anti-TNF-alpha mAb treatment completely abolished the antitumor effect of LC 9018 in vivo, suggesting that in this model LC 9018 has a survival-prolonging effect involving certain cytokines. Moreover, i.pl. injection of mouse recombinant TNF-alpha into Meth A-bearing mice pretreated with anti-TNF alpha mAb partially restored the survival-enhancing effect of LC 9018. These results led us to conclude that TNF-alpha induced by i.pl. injection of LC 9018 plays an important role in the antitumor effect of LC 9018 in vivo. PMID- 10691089 TI - Rat dorsal root ganglia neurons as a model for Listeria monocytogenes infections in culture. AB - Neurotropism of Listeria monocytogenes was studied in rat dorsal root ganglia (DRG) and hippocampal neurons in culture. Using a system in which the DRG neurons can grow relatively free from other cells, it was observed that such DRG neurons, in contrast to hippocampal neurons, can be effectively infected by L. monocytogenes. The bacteria aligned along DRG axons, but not along hippocampal neurites. A mutant deficient in internalin, a protein required for entry into E cadherin-expressing cells, did not interact with DRG neurons. Axonal migration of bacteria was studied in the DRG neurons grown in a double-chamber system, where either the neurites or the nerve cell bodies were exposed to the bacteria. The data suggest that L. monocytogenes can infect both axons and DRG nerve cell bodies, and that the bacteria can migrate in a retrograde as well as anterograde direction. These results support the notion that L. monocytogenes can spread via primary sensory neurons to the central nervous system. Infection of DRG primary sensory neurons, as employed in the present study, provides a model for analysis of bacterial and neuronal factors of importance for neurovirulence of L. monocytogenes. PMID- 10691090 TI - A membrane-located glycosphingolipid of monocyte/granulocyte lineage cells induces growth arrest and triggers the lytic viral cycle in Epstein-Barr virus genome-positive Burkitt lymphoma lines. AB - Gangliosides are known to influence cell growth and differentiation. The neolacto series ganglioside IV3NeuAc-nLc4 (2-->3-sialosylparagloboside) is present in members of the monocyte/granulocyte lineage, but is not found in cells that belong to the lymphocyte lineage. In this study we demonstrated that IV3NeuAc nLc4 inhibits the proliferation of Epstein-Barr virus (EBV) genome-positive Burkitt lymphoma cells of the lines Raji and P3HR-1K. IV3NeuAc-nLc4-induced growth inhibition is associated with an increase in G0/G1 phase cells and a reduced expression of CD21 and HLA-DR antigens on Raji cells. These data suggest that IV3NeuAc-nLc4 may affect differentiation of lymphoma cells. Additionally, the increased expression of viral mRNA species which are characteristic for the lytic viral cycle in the non-producer line Raji and the enhanced release of virions from the producer line P3HR-1K demonstrate that IV3NeuAc-nLc4 activates the replication of EBV. Growth inhibition and termination of the viral latency suggest that IV3NeuAc-nLc4 present in monocyte/granulocyte lineage cells may be an effector of the natural defense against EBV persistency and transformation. PMID- 10691091 TI - The ferric uptake regulator (Fur) homologue of Helicobacter pylori: functional analysis of the coding gene and controlled production of the recombinant protein in Escherichia coli. AB - A homologue of the ferric uptake regulator protein Fur has recently been identified within the Helicobacter pylori genome. The promoterless gene on a plasmid did partially complement a fur-negative mutant of Escherichia coli, and was strongly positive in the Fur titration assay (FURTA). The genetic and functional characterization of the complete fur homologue performed in this study revealed that the gene is conserved among H. pylori strains ( > 95% identity), and does not carry nucleotide transitions in iron-resistant mutants of H. pylori. The fur homologue on a plasmid mediated full iron-dependent ferric uptake regulator activity in the fur-deficient mutant strains H1681 and H1780 of E. coli. Immunoblot analysis revealed that Fur from H. pylori cross-reacts with antibodies raised against Fur from E. coli. The fact that inactivation of the fur gene abolished the FURTA-positive phenotype in the E. coli indicator strain H1717, indicated that this phenotype is rather caused by the encoded protein than by real Fur titration. Subcloning of the fur gene into an expression vector allowed controlled production in E. coli, and purification of a recombinant version of the H. pylori Fur protein. In summary, the results confirm the function of the H. pylori Fur homologue as iron-dependent transcriptional repressor by its ability to interact with the Fur-regulated promoters of the genes fiu and fhuF in E. coli. PMID- 10691092 TI - Influence of the genetic background on the pattern of lesions developed by resistant and susceptible mice infected with Paracoccidioides brasiliensis. AB - To compare the sequential evolution of lesions developed by resistant (A/Sn) and susceptible (B10.A) mice to Paracoccidioides brasiliensis infection we inoculated a virulent isolate of the fungus and collected the pancreas/peripancreatic omentum monthly (from 1 to 6 months) post infection. After fixation, tissue sections were stained by conventional methods for light microscopy to investigate the cellular composition, the extracellular matrix (ECM) patterns and the morphology of the yeasts in the lesions. In both strains, the fungal lesions were localized mostly in the omentum; a few lesions in the pancreatic parenchyma were observed, mostly in B10.A mice. In both strains, macrophages and plasmocytes were the predominant cells in all lesions, followed by neutrophils (PMN) and macrophages transformed into giant and epithelioid cells. Remarkable differences were observed between resistant and susceptible mice, specially related to the ECM structure of the granulomatous lesions. In A/Sn mice, from the 1st month on, the coexistence of two types of lesions was observed: one type showed a well defined encapsulated nodule, constituted mainly of type I collagen. Neutrophils were abundant in areas of massive fungal destruction and few viable yeasts were observed. The other type showed residual characteristics, with sparse collagen deposits and presence of xantomatous-like macrophages, containing degenerated fungi. Such residual lesions predominated after the 2nd month and were the only type observed from the 4th month on, indicating the control of the infection. In B10.A mice, on the contrary, only one type of lesion was observed, showing less tendency to encapsulation and the formation of multiple small granulomatous foci, individualized by reticular type III collagen fibers. There were many plasmocytes in the periphery and large numbers of budding yeasts, with no evidence of fungal destruction. In the course of the infection the lesions progressively increased in number and size. Altogether, the comparative histopathological analysis demonstrates the influence of the genetic pattern of the host on the lesions developed by resistant and susceptible mice to P. brasiliensis infection. PMID- 10691093 TI - Hantavirus Dobrava infection with pulmonary manifestation. AB - Dobrava virus infection was diagnosed serologically by enzyme-linked immunosorbent and immunofluorescence assays. To determine which hantavirus serotype was involved, sera were analyzed by a focus reduction neutralization test. The clinical data indicated that only pulmonary manifestation was present. Our data support the presence of Dobrava virus infection outside the Balkan region. In conclusion, a previously healthy adult with unexplained pulmonary perfusion failure should be investigated for hantavirus infection. PMID- 10691094 TI - Soluble fibrin species in arterial thrombi. AB - The aim of this study was to characterize soluble fibrin(ogen) species in human, arterial, in-vivo-formed thrombi, using the immunoblotting technique. Specimens were collected via intra-arterial catheters in six patients scheduled for catheter-directed thrombolysis. Unreduced and reduced samples of the supernatants from the arterial thrombi-derived specimens were electrophoresed on polyacrylamide gels and immunoblotted, using specific mono- and polyclonal anti fibrin(ogen) antibodies. The reduced samples disclosed substantial amounts of high molecular weight material, consistent with alpha-chain polymers and gammagamma-dimers, as well as lower molecular weight material, such as alpha-, beta- and gamma-chains. No fibrinogen with intact fibrinopeptide A was detectable, and des-AABB fibrin represented a major fibrin derivative in the soluble part of the arterial thrombi. The alpha-chains were C-terminally degraded, most of them distal to position 259. In conclusion, we have demonstrated the presence of cross-linked fibrin derivatives in soluble, arterial thrombus-related material, without signs of fibrinogen-fibrin hybrids. The fibrin derivatives were C-terminally degraded, thus representing X-oligomeric material, most probably originating from plasmin degradation of insoluble thrombus fibrin. The present study supports the hypothesis of a dynamic equilibrium between clotting and lysis in thrombi. PMID- 10691095 TI - Absorption and antithrombotic activity of unfractioned heparin after intraduodenal administration in rats. AB - In the search for a more acceptable route of heparin administration that can also be used for long-term treatment, we evaluated the bioavailability and antithrombotic activity of intraduodenal administration of unfractioned heparin (UFH) in rats. A radioiodinate derivative of UFH was administered intraduodenally in rats in conjunction with unlabeled UFH. We found that radioactivity increases very rapidly in plasma, as well as on the surface of aortic and caval segments, so that peak radioactivity was already achieved within 5 min of drug administration. Subsequently, plasma radioactivity declined rapidly, although 1.5 2.5% of the total radioactivity administered was still circulating 3 h after drug administration. The plasma anti-Xa activity was much lower and longer lasting than expected from the radioactivity counts in both peripheral and portal blood, and its level was very similar in the two circulatory districts, never exceeding 023 U/ml. This suggests that extensive degradation of the drug already occurs during its gastrointestinal absorption. Nevertheless, in a stasis-induced venous thrombosis model, intraduodenal UFH prevented thrombus formation in a dose dependent way (ED50, 2000 IU/kg). The maximum antithrombotic effect was observed when the drug was administered 30-60 min before ligature of the vena cava, and a 40% reduction of thrombus weight was still present at 180 min. Since antithrombotic kinetics does not match the kinetics of either the plasma or vessel-bound radioactivity but approaches what is found in anti-Xa activity, the antithrombotic activity of oral heparin may be dependent on the release of unlabeled endogenous glycosaminoglycans deposited in the vessels. PMID- 10691096 TI - Characterization of immortalized human umbilical and iliac vein endothelial cell lines after transfection with SV40 large T-antigen. AB - Most in vitro studies of human endothelial cells have relied on cells derived from human umbilical veins (HUVEC); however, heterogeneity of primary cultured endothelial cells can make critical interpretation of results difficult. Several endothelial cell lines have been produced to serve as a more constant source of endothelial cells. In this study, we characterized the endothelial cell lines EVLB3 and EVLC2 derived from HUVEC, and EVLK1 and EVLK2 derived from human iliac vein endothelial cells (HIVEC). These cell lines maintained the typical endothelial cell cobblestone morphology and appeared to be growth factor independent. They lost PECAM-1 and von Willebrand factor, GP96 was reduced to the level of vascular smooth muscle cells (SMC), but aSMC-actin was far less than in vascular SMC. Antigen levels of tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAI-1) were comparable with young endothelial cells, and mRNA was present for tPA, PAI-1, tissue factor (TF), tissue factor pathway inhibitor and thrombomodulin. This study revealed that mRNA and protein expression of coagulation and fibrinolytic factors was influenced by the stage of cell confluence. No differences could be detected between the endothelial cell lines derived from HUVEC and HIVEC. These cell lines may be a useful tool for studies on cellular interactions of fibrinolytic components or exploring the regulation of TF expression. PMID- 10691097 TI - Use of phage display for the generation of human antibodies that neutralize factor IXa function. AB - The use of libraries of phage-displayed human single-chain antibody fragments (scFv) has become a new, powerful tool in rapidly obtaining therapeutically useful antibodies. Here, we describe the generation of human scFv and F(ab')2 directed against the gamma-carboxyglutamic acid (Gla) domain of coagulation factor IX. A large library of human scFv, displayed either on M13 phage or expressed as soluble proteins, was screened for binding to human Gla-domain peptide (Tyr1-Lys43). Among a panel of scFv that bound to the factor IX-Gla domain, six scFv clones recognized full-length factor IX and exhibited strong inhibitory activity of factor IX in vitro. After reformatting as F(ab')2, the affinity for factor IX of three selected clones was determined: 10C12 Kd = 1.6 nmol/l, 13D1 Kd = 2.9 nmol/l, and 13H6 Kd = 0.46 nmol/l. The antibodies specifically bound to factor IX and not to other coagulation factors, as assessed by enzyme-linked immunosorbent-type and human plasma clotting assays. The complementarity determining region amino acid sequences of clones 10C12 and 13D1 only differed at a single residue, whereas 13H6 showed little homology, suggesting that 13H6 binds to a different epitope within the factor IX-Gla domain. Despite the slightly lower affinity of 10C12 F(ab')2 versus 13H6 F(ab')2, 10C12 was consistently more potent than 13H6 in prolonging the activated partial thromboplastin time (APTT), in inhibiting platelet-mediated plasma clotting, and in inhibiting factor X activation by the intrinsic Xase complex. Finally, 10C12 F(ab')2 also recognized and neutralized factor IX/factor IXa of different species, as demonstrated by the specific APTT prolongation of dog, mouse, baboon and rabbit plasma. In summary, the results validate the usefulness of scFv phage displayed libraries to rapidly generate fully human antibodies as potential new therapeutics for thrombotic disorders. PMID- 10691098 TI - Plasma fibrinogen, haemostatic factors and prediction of peripheral arterial disease in the Edinburgh Artery Study. AB - The role of fibrinogen and other haemostatic factors in prediction of peripheral arterial disease (PAD) has not been established. We examined the associations of plasma fibrinogen, von Willebrand Factor (vWF), tissue plasminogen activator (t PA) antigen, fibrin D-dimer, and factor VII with the development and clinical progression of PAD. In the Edinburgh Artery Study, 1592 men and women, aged between 55 and 74 years, were followed prospectively over 5 years to detect the onset of PAD, and the deterioration of established PAD. At baseline, 418 individuals had evidence of PAD and 60 (14.4%) subsequently deteriorated. 1080 subjects had no baseline disease, but 59 (5.5%) developed PAD during follow-up. Median levels of fibrinogen and vWF were higher in the group developing disease compared with the group which did not (2.78 g/l versus 2.57 g/l, P< or =0.01; 116 IU/dl versus 104 IU/dl, P< or =0.05; respectively). After adjusting for age and sex, fibrinogen (P< or =0.01) and vWF (P< or =0.05) were significantly associated with the risk of developing PAD. The association between fibrinogen and development of disease remained after adjusting for cardiovascular risk factors and baseline ischaemic heart disease (relative risk, 1.35, 95% confidence interval, 1.05, 1.73; P< or =0.05). None of the haemostatic factors were significantly associated with progression of PAD. In conclusion, plasma fibrinogen levels are related to the future onset of PAD, providing further evidence of a possible role of elevated fibrinogen in the development of atherosclerotic disease. PMID- 10691099 TI - Pharmacologic modulation of thrombin generation associated with human clots by human purified antithrombin alone or in the presence of low molecular weight heparin or unfractionated heparin. AB - Procoagulant activities associated with human clots may contribute to thrombus extension. We investigate the inhibition of clot-associated factor Xa and thrombin activities by purified human antithrombin either alone or as combination with a low molecular weight heparin (enoxaparin) as compared with unfractionated heparin (UFH). The standard clots were prepared by recalcification of frozen platelet-poor human plasma. Clot-associated thrombin was measured on the clot after clot incubation in recalcified buffer or recalcified prothrombin solution. The enzymatic reaction was measured using a specific substrate for thrombin (CBS 3447). The thrombin concentration was determined both on the clots and in the reaction mixtures. In parallel, prothrombin fragment 1.2 and thrombin antithrombin complexes (TAT) were measured using enzyme-linked immunosorbent assay methods. We demonstrated that in the presence of purified human prothrombin and antithrombin (AT), a partial inhibition of clot associated thrombin activity correlated with an increase of TAT complexes. However, antithrombin was unable to inhibit thrombin generation induced by the clot-associated factor Xa. Enoxaparin (low molecular weight heparin) and UFH did not enhance clot-bound thrombin inhibition induced by AT. We conclude that clot-bound thrombin is accessible to human antithrombin alone. AT is also able to inhibit thrombin generated by factor Xa-associated clot. However, neither a low molecular weight heparin or UFH enhanced the effect of AT alone. PMID- 10691100 TI - Platelet activation during angiotensin II infusion in healthy volunteers. AB - The present study was undertaken to evaluate the effects of an intravenous infusion of angiotensin II (10 ng/kg per min) on platelet function and coagulation in vivo in 18 healthy males. The infusion increased mean arterial pressure by 23+/-2 mm Hg. Plasma beta-thromboglobulin levels, reflecting platelet secretion, increased by 66+/-24% during the infusion, as did also platelet surface expression of P-selectin as measured by flow cytometry. Platelet fibrinogen binding increased, whereas platelet aggregability, assessed by ex vivo filtragometry, was unaltered. Angiotensin II caused mild activation of the coagulation cascade with increases in plasma levels of thrombin-antithrombin complex and prothrombin fragment F1 + 2. In conclusion, angiotensin II has mild platelet-activating effects in vivo and also enhances coagulation. Markers of platelet secretion are significantly increased, whereas markers of platelet aggregability are less affected. The clinical relevance of these findings remains to be clarified. PMID- 10691101 TI - Plasma viscosity, fibrinogen and the metabolic syndrome: effect of obesity and cardiorespiratory fitness. AB - The association between both plasma viscosity and fibrinogen concentration with clustering of metabolic risk markers was examined within a cross-sectional study of employed middle-aged men. Analyses were performed on a subsample of 629 non smokers (46.7+/-7.8 years) without diabetes. The effect of obesity and cardiorespiratory fitness on these haemorheological parameters and their association with the metabolic syndrome was also investigated. The cohort was grouped by the number of metabolic markers present. Metabolic markers included high-density lipoprotein-cholesterol (<1.13 mmol/l), triglycerides (> or =1.805 mmol/l), glucose (> or =5.5 mmol/l) and diastolic blood pressure (> or =90 mm Hg). The age-adjusted odds ratio for hyperviscosity (> or =1.67 mPa/s) was 2.08 [95% confidence interval (CI), 1.06-4.05; P = 0.031] for the subjects with the metabolic syndrome (three or more metabolic markers) when compared with those with no metabolic abnormalities. The comparable age-adjusted odds ratio for hyperfibrinogenaemia (> or = 3.47 g/l) was non-significantly higher at 1.69 (95% CI, 0.87-3.27; P = 0.119). The mean age-adjusted plasma viscosity level and the prevalence of hyperviscosity increased significantly from 1.629 to 1.692 mPa/s (P = 0.0005) and from 21.0 to 36.0% with accumulating metabolic markers (P = 0.006). Plasma viscosity and fibrinogen concentration both increased with higher quartiles of skinfolds (P = 0.003 and P = 0.01, respectively) following adjustment for age, lipids and leucocyte count. Plasma viscosity was also significantly lower with higher levels of predicted maximum oxygen consumption (VO2max) (P = 0.0005). The odds ratio for hyperviscosity in subjects with the metabolic syndrome as compared with those with no metabolic markers was attenuated following adjustment for age, sum of skinfolds and predicted maximum oxygen consumption (VO2max) (1.44; 95% CI, 0.72-2.90; P = 0.307). These cross sectional results suggest that plasma viscosity is associated with increased clustering of metabolic markers in middle-aged men of high socio-economic status. Obesity and poor cardiorespiratory fitness may be important in the development of haemorheological abnormalities associated with the metabolic syndrome. PMID- 10691102 TI - A kinetic model of the circulatory regulation of tissue plasminogen activator during orthotopic liver transplantation. AB - To better understand the regulation of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) during liver transplantation, we used a computer model of the human circulatory system to simultaneously evaluate the effect of t-PA secretion, t-PA inhibition by PAI-1, hepatic clearance of t-PA, blood loss, transfusion and hemodynamics on t-PA and PAI-1 levels during liver transplantation in three patients that differed in severity of liver disease, blood loss and anhepatic changes in t-PA. Higher preoperative t-PA levels were primarily related to underlying liver disease and reduced hepatic clearance. During the anhepatic stage, when hepatic t-PA clearance was eliminated: (1) the expected rise in t-PA was modulated by the extent of bleeding, which acted as an alternate t-PA clearance mechanism; and (2) the ratio of t-PA:PAI-1 was increased due both to lower t-PA clearance and reduced PAI-1 secretion. Recirculation of the new liver was associated with renewed clearance of t-PA, an acute phase increase in PAI-1 and a drop in the t-PA:PAI-1 ratio. Understanding fibrinolytic regulation required simultaneous analysis of t-PA secretion, inhibition and clearance. Anhepatic t-PA levels could be predicted based on preoperative liver function and surgical blood loss, which acted as an alternate t-PA clearance mechanism. PMID- 10691103 TI - Mutations in a potential phospholipid binding loop in the C2 domain of factor V affecting the assembly of the prothrombinase complex. AB - Activated factor V (FVa) serves as a cofactor to activated factor X in the prothrombinase complex. FVa is homologous to activated factor VIII (FVIIIa), the light chains of both proteins being formed by similar domains (A3-C1-C2). Interaction of FVa and FVIIIa with negatively charged phospholipid membranes is crucial for the function of both cofactors. In both proteins, the C2 domains are important for membrane binding but a detailed understanding of the binding mechanisms is missing. Recently, knowledge has been gained into the three dimensional structures of the C domains facilitating studies of structure function relationships. Structural analysis of the C2 domain in FVa predicted a surface-exposed loop (K2060, K2061, S2062, W2063, W2064) to be involved in membrane binding. Three double mutants were created, K2060Q-K2061Q, W2063Y-W2064Y and W2063A-W2064A, and expressed in a transient expression system. In addition, a FV variant in which all four residues were mutated, K2060Q-K2061Q-W2063A-W2064A, was produced. Mutagenesis of the two lysines showed no functional consequences, whereas mutagenesis of the two tryptophanes yielded FVa with impaired ability to interact with the phospholipid, as demonstrated by a poor functional activity at limiting phospholipid concentrations. A molecular model of FVa, anchored at the surface of a phospholipid membrane, was developed and used as a template for the interpretation of the mutagenesis experiments. PMID- 10691104 TI - Deep venous thrombosis prophylaxis in trauma: cost analysis. AB - Deep vein thrombosis and pulmonary embolism are major risks in patients experiencing major trauma. Currently, the American College of Chest Physicians recommends low molecular weight heparin as prophylaxis in trauma patients with identifiable risk factors in the absence of contraindications. Enoxaparin is the only low molecular weight heparin available in the US that has been evaluated to date in this indication. The purpose of this study was to perform incremental cost-effectiveness ratio calculations for enoxaparin versus no prophylaxis as thromboembolic prophylaxis in trauma patients. These calculations demonstrate that a cost of $279.43 would be incurred for each thromboembolic event avoided if enoxaparin 30 mg every 12 h were routinely used as prophylaxis in this population, compared with no prophylaxis. Sensitivity analyses demonstrate that if the incidence of proximal vein thrombosis in patients prophylaxed with enoxaparin approached 1.8%, if the actual rate of these thrombi exceeded 19.4% in untreated patients, or if the cost of the drug was decreased to $15.25 per dose, a cost saving would be experienced in routine prophylaxis with this agent. PMID- 10691105 TI - Right atrial and ventricular thrombi in Behcet's disease: a case report and review of literature. AB - Behcet's disease is a chronic multi-system disease presenting with recurrent oral and genital ulceration, and relapsing uveitis. Cardiac involvement is an extremely rare manifestation of this disorder. We report an unusual case of Behcet's disease characterized by a mural cardiac thrombi in the right atrium and right ventricle along with transient protein C and S deficiency. PMID- 10691106 TI - Guinea pig platelets do not respond to GYPGKF, a protease-activated receptor-4 activating peptide: a property distinct from human platelets. PMID- 10691107 TI - Felbamate in experimental model of status epilepticus. AB - PURPOSE: To examine the putative seizure-protective properties of felbamate in an animal model of self-sustaining status epilepticus (SSSE). METHODS: SSSE was induced by 30-min stimulation of the perforant path (PPS) through permanently implanted electrodes in free-running male adult Wistar rats. Felbamate (FBM; 50, 100, and 200 mg/kg), dizepam (DZP; 10 mg/kg), or phenytoin (PHT; 50 mg/kg) were injected i.v. 10 min after SSSE induction. Electrographic manifestations of SSSE and the severity of SSSE-induced neuronal injury were analyzed. RESULTS: Felbamate injected during the early stages of SSSE (10 min after the end of PPS), shortened the duration of seizures in a dose-dependent manner. Total time spent in seizures after FBM and 290 +/- 251 min (50 mg/kg), 15.3 +/- 9 min (100 mg/kg), and 7 +/- 1 min (200 mg/kg), whereas control animals spent 410 +/- 133 min seizing. This effect of FBM was stronger than that of DZP (10 mg/kg, 95 +/- 22 min) and comparable to that of PHT (50 mg/kg, 6.3 +/- 2.5 min). In the applied doses, FBM (200 mg/kg) was more effective than PHT (50 mg/kg) or DZP (10 mg/kg) in shortening seizure duration and decreasing spike frequency, when administered on the pleateau of SSSE (injection 40 min after the end of PPS). Anticonvulsant action of FBM was confirmed by milder neuronal injury compared with control animals. CONCLUSIONS: Felbamate, a clinically available AED with a moderate affinity for the glycine site of the NMDA receptor, displayed a potent seizure protective effect in an animal model of SSSE. These results suggest that FBM might be useful when standard AEDs fail in the treatment of refractory cases of SE. PMID- 10691108 TI - Effects of vigabatrin on sleep-wakefulness cycle in amygdala-kindled rats. AB - PURPOSE: Our aim was to study the effect of prolonged administration of vigabatrin (VGB) on sleep-wakefulness cycle in kindled seizure-induced rats. METHODS: Adult male Wistar rats were implanted stereotaxically with electrodes for kindling and polysomnography. The rats were divided into two groups, kindled and VGB-treated kindled rats. VGB was administered intraperitonially every day for 21 days, and polysomnographic recordings were taken after doses 1, 7, 14, and 21. The drug effects were evaluated by comparing the records of kindled and drug treated kindled rats. RESULTS: The VGB-administered kindled rats showed an increase in total sleep time (TST) due to an increase in total non-rapid eye movement (NREM) and light slow-wave sleep stage I (SI) with a decrease in wakefulness. The number of episodes and REM onset latencies were found to be decreased after drug treatment. CONCLUSIONS: It can therefore be concluded that VGB has a somnolence-inducing effect and that it might mediate its anticonvulsant effect by altering sleep architecture through sleep-regulating areas. PMID- 10691109 TI - Evidence favoring genetic heterogeneity for febrile convulsions. AB - PURPOSE: Two large Canadian kindreds appearing to segregate febrile convulsions as an autosomal dominant trait were evaluated for linkage to three known FC loci, as well as other epilepsy loci. METHODS: Members of the two families were genotyped with microsatellite markers linked to the previously identified febrile convulsion loci, FEB1, FEB2, and GEFS+, and we performed two-point linkage analyses by assuming an autosomal dominant mode of inheritance. RESULTS: We report the exclusion of the FC trait in our families to FEB1 on 8q13-21 and to a second febrile convulsion locus on 19p13. Furthermore, we also excluded the GEFS+ locus on 19q13.1 as the cause of febrile convulsions in both kindreds. Microsatellite markers linked to juvenile myoclonic epilepsy (EJM1), benign neonatal familial convulsions EBN1 and EBN2, autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), idiopathic generalized epilepsy (EGI), progressive myoclonic epilepsy of Unverricht-Lundborg (EPM1), and partial epilepsy with auditory features (EPT), were also excluded as potential loci linked to the FC trait in our families. CONCLUSIONS: These findings favor considerable genetic heterogeneity for febrile convulsions. PMID- 10691110 TI - Expression of 5alpha-reductase and 3alpha-hydroxisteroid oxidoreductase in the hippocampus of patients with chronic temporal lobe epilepsy. AB - PURPOSE: The hippocampus is one of the principal target areas for neurosteroidal action, and the major neuroendocrine conversion of progesterone appears to be 5alpha-reduction and 3alpha-hydroxysteroid oxidoreduction, leading to the potent neurosteroid 3alpha,5alpha-tetrahydroxyprogesterone. To investigate whether the human hippocampus is equipped with the enzymes 5alpha-reductase and 3alpha hydroxysteroid oxidoreductase (3alpha-HSOR), we studied the expression of 5alpha reductase types 1 and 2 and 3alpha-HSOR types 1 and 2 in the resected hippocampi of patients with medically intractable chronic temporal lobe epilepsy. METHODS: We studied tissue specimens from the hippocampi of 13 women, 25 men, and four children. Quantification of different mRNAs was achieved by competitive reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: 5Alpha-reductase 1 mRNA and 3alpha-HSOR 2 mRNA were expressed in hippocampi of children and adults, whereas 5alpha-reductase 2 mRNA and 3alpha-HSOR 1 mRNA were not expressed. Neither 5alpha-reductase 1 mRNA nor 3alpha-HSOR 2 mRNA concentrations in hippocampal tissue showed any statistically significant differences between women and men or between children and adults. CONCLUSIONS: This study demonstrates for the first time mRNA expression of the type 1 isozyme of 5alpha-reductase and the type 2 isozyme of 3alpha-HSOR in the human hippocampus. The finding that both 5alpha-reductase and 3alpha-HSOR are present in the hippocampus leads us to assume the synthesis of neuroactive steroids in this human brain area. PMID- 10691111 TI - The potential for vigabatrin-induced intramyelinic edema in humans. AB - PURPOSE: Vigabatrin (Sabril, Hoechst Marion Roussel) is an antiepilepsy drug (AED) presently marketed in 64 countries for the treatment of partial and secondarily generalized seizures. Vigabatrin (VGB) is marketed in a subset of these countries for the treatment of infantile spasms. Clinical experience in humans has shown that VGB provides effective seizure control with a wide margin of safety. However, animal toxicity studies raised concern when prolonged administration of VGB was shown to induce intramyelinic edema (IME) in some laboratory animal species. METHODS: Animal and human data were reviewed with respect to the potential for VGB-induced IME. Surveillance of patients receiving VGB in clinical trials or by prescription has been conducted for >15 years to identify patients developing clinical abnormalities that might be IME related. RESULTS: The histologic lesions of VGB-induced IME in animals are reliably reproduced and correlate with changes in multimodality evoked potentials (EPs) and magnetic resonance imaging (MRI). Numerous studies of the effects of VGB on EP and MRI in epilepsy patients have demonstrated no clear-cut IME-related changes in these modalities. Additionally, autopsy and surgical brain samples from VGB-treated patients have been scrutinized for potential IME histopathology. In an estimated 350,000 patient-years of VGB exposure (approximately 175,000 patients exposed for 2 years at an average dose of 2 g/day), no definite case of VGB-induced IME has been identified. CONCLUSIONS: Comprehensive review of a variety of sources of data failed to identify any definite case of IME in humans treated with VGB. PMID- 10691112 TI - Subacute electrical stimulation of the hippocampus blocks intractable temporal lobe seizures and paroxysmal EEG activities. AB - PURPOSE: To investigate the clinical, electroencephalographic (EEG), and histopathologic effects of subacute electrical stimulation of the hippocampal formation or gyrus (SAHCS) on 10 patients with intractable temporal lobe seizures. METHODS: Bilateral, depth, hippocampal or unilateral, subdural, basotemporal electrodes were implanted in all 10 patients for a topographic diagnosis of the site and extent of the epileptic focus before a temporal lobectomy. In all patients, antiepileptic drugs (AEDs) were discontinued from 48 to 72 h before a program of continuous SAHCS, which was performed for 2-3 weeks. Stimulation parameters were biphasic Lilly wave pulses, 130/s in frequency, 450 micros in duration, and 200-400 microA in amplitude. The stimuli were delivered 23 of every 24 h for the 2-3-week SAHCS period. The effects of SAHCS on the number of clinical seizures per day and the percentage of interictal EEG spikes per 10-second samples of maximal paroxysmal activity at the epileptic focus were determined daily during the 16 days of SAHCS. At the completion of this program, patients underwent an en bloc temporal lobectomy, and the histopathologic effects of SAHCS on the stimulated tissue were analyzed by means of light-microscopy studies. RESULTS: In seven patients whose stimulation electrode contacts were placed within the hippocampal formation or gyrus and who experienced no interruption in the stimulation program, SAHCS abolished clinical seizures and significantly decreased the number of interictal EEG spikes at the focus after 5 6 days. The most evident and fast responses were found by stimulating either the anterior pes hippocampus close to the amygdala or the anterior parahippocampal gyrus close to the entorhinal cortex. Other surface, hippocampal, and basotemporal EEG signs predicted and accompanied this antiepileptic response. These included an electropositive DC shift and monomorphic delta activity at the medial hippocampal and parahippocampal regions, and a normalization of the background EEG activity and signs of slow-wave sleep in surface. depth, and subdural regions. In contrast, no evident antiepileptic responses or no responses at all were found in three patients when stimulation was either interrupted or when it was administered outside the hippocampus. Light microscopy analysis of the stimulated hippocampal tissue showed histopathological abnormalities attributable to the depth-electrode penetration damage or to the pial surface reaction to the subdural, Silastic electrode plate. However, no evident histopathological differences were found between the stimulated and nonstimulated hippocampal tissue. CONCLUSIONS: SAHCS appears to be a safe procedure that can suppress temporal lobe epileptogenesis with no additional damage to the stimulated tissue. PMID- 10691113 TI - The impact of a single seizure on health status and health care utilization. AB - PURPOSE: To assess the health status of patients after a single seizure. METHODS: We compared single-seizure patients (SS) with patients who had well-controlled epilepsy (WC), and uncomplicated hypertension (HT). Patients were adults screened from emergency and outpatient units of two urban teaching hospitals using predefined criteria. The 83 patients (SS, 30; WC, 29; HT, 24) were interviewed by phone about functional status (SF-36), comorbid illness, cause of illness, number of visits to health providers, and drug side effects. RESULTS: No significant differences were found among groups for health status, SF-36 domain, or occurrence of drug side effects. SS patients had significantly lower scores on vitality (p < 0.03) and a trend toward lower role physical function (p < 0.07) compared with age-adjusted population norms. SS reported more visits to health providers than WC or HT, and the number of visits remained high at interview 1 year later. Patient knowledge of the "reason" for the seizure was not associated with health status or number of visits. CONCLUSIONS: Health status of patients within 1 year of a single seizure is similar to that of patients with well controlled epilepsy or hypertension, but SS patients have greater health care utilization. PMID- 10691114 TI - Moderators of the effect of preoperative emotional adjustment on postoperative depression after surgery for temporal lobe epilepsy. AB - PURPOSE: Other outcome measures besides seizure control must be considered when assessing the benefit of epilepsy surgery. We investigated the effect of preoperative psychosocial adjustment on postoperative depression in epilepsy patients followed up prospectively for 2 years after temporal lobectomy. METHODS: The Washington Psychosocial Seizure Inventory (WPSI) evaluated psychosocial functioning; the Centre for Epidemiological Studies Depression Scale (CES-D) measured depression. Both were completed at baseline and follow-up. RESULTS: Follow-up occurred in 39 temporal lobectomy patients at 2 years after surgery. Greatest improvement in depression scores was limited to patients with good seizure outcomes (seizure free, or marked reduction in seizure frequency), and seizure outcome was a significant predictor of postoperative depression. Despite this, preoperative scores on the emotional adjustment scale of the WPSI were most highly correlated with depression 2 years after surgery. To clarify this relation, moderated hierarchic regression suggested that good preoperative emotional adjustment (WPSI) was generally associated with less depression after surgery. Moreover, poorer preoperative adjustment combined with older age, generalized seizures, the finding of preoperative neurologic deficits, a family history of psychiatric illness, and/or a family history of seizures was related to higher depression scores 2 years after surgery. CONCLUSIONS: Depression after temporal lobectomy is dependent on a complex interaction of variables and can have a significant effect on indices of postoperative adjustment. The WPSI emotional adjustment scale may help to predict which patients are likely to be chronically depressed after surgery. PMID- 10691115 TI - Induction of partial epileptic seizures by flumazenil. AB - PURPOSE: This study addressed the efficacy of flumazenil (FMZ) to induce or activate interictal or ictal epileptic discharges in patients with medically intractable partial epilepsies. METHODS: Flumazenil, 1 mg, was injected intravenously in 67 patients undergoing presurgical monitoring for epilepsy surgery, 49 of whom had been treated with benzodiazepines (BZDs) before flumazenil was given. Continuous video electroencephalogram (EEG) monitoring with surface or intracranial electrodes was used to evaluate interictal EEG activity, ictal discharges, and the occurrence and semiology of clinically manifest epileptic seizures. RESULTS: Interictal epileptiform potentials did not change in frequency or distribution after FMZ. In patients not pretreated with BZDs, epileptic seizures could not be provoked. In eight of the 49 patients pretreated with BZDs, epileptic seizures occurred within 30 min of FMZ application. Seizure semiology and regional EEG onset were identical to seizures recorded without FMZ. Patients operated on according to seizure-onset localization with FMZ had a >75% reduction in seizure frequency or became seizure free. CONCLUSIONS: Seizure induction by FMZ seems to be a valid method for evaluating seizure semiology and localization of the seizure-onset zone during presurgical monitoring of patients with medically intractable localization-related epilepsies. PMID- 10691116 TI - Planned ictal FDG PET imaging for localization of extratemporal epileptic foci. AB - PURPOSE: This work demonstrates the feasibility of planned ictal positron emission tomography (PET) with [18F]fluoro-2-deoxy-glucose (FDG) for localization of epileptic activity in patients with frequent partial seizures of extratemporal origin. METHODS: Ictal PET imaging was performed in four patients (two men and two women, ages 28-61) with continuous or very frequent (every 3-15 min) partial seizures. All patients had abnormalities apparent on magnetic resonance (MR) or computed tomographic (CT) imaging, two with extensive brain lesions that precluded precise localization of the seizure focus with interictal PET or single photon emission tomography (SPECT) imaging. RESULTS: Ictal PET imaging demonstrated a restricted area of focal hypermetabolism concordant with surface electroencephalographic (EEG) recording in all cases. The PET images were registered to MR imaging data for further anatomic localization of hypermetabolic regions in three cases. The ictal PET data were used to guide neurosurgical intervention in one case. CONCLUSIONS: We conclude that planned ictal PET imaging may be a useful and potentially superior approach to ictal SPECT for identifying the epileptic focus in a selected group of patients with continuous or frequent simple partial seizures. PMID- 10691117 TI - Spindles-inducing mechanism modulates sleep activation of interictal epileptiform discharges in the Landau-Kleffner syndrome. AB - PURPOSE: Landau-Kleffner syndrome (LKS) is characterized by a marked increase of interictal epileptiform discharges (IEDs) during sleep. During nonrapid eye movement (NREM) sleep, neuronal membrane potential oscillations lead to the appearance of spindles and delta waves in the surface EEG and might develop into paroxysmal synchronization. Spectral analysis allows the quantitative description of the dynamics of delta (slow-wave activity, SWA, 0.5-4.5 Hz) and sigma activity (SA, 12.0-16.0 Hz) and can be used to assess the relation between SA, SWA, and IEDs during sleep. METHODS: We performed six overnight continuous EEG polysomnographic studies in three patients with LKS. The temporal series of SWA and SA were obtained from a spike-free derivation lead. The IEDs count was performed on the most active lead. Relations between sigma and SWA and time series of IEDs were tested by means of correlation techniques after data normalization. RESULTS: Our results revealed a significantly higher correlation between IEDs and SA with respect to SWA in all the subjects, in total sleep time. The same analysis limited to NREM sleep highlights the better correlation between SA and IEDs. CONCLUSIONS: Our data suggest that neural mechanisms involved in the generation of sleep spindles facilitate IEDs production in LKS. PMID- 10691118 TI - Contralateral EEG slowing and amobarbital distribution in Wada test: an intracarotid SPECT study. AB - PURPOSE: To relate the occurrence of contralateral electroencephalogram slowing (CES) to amobarbital distribution, we performed electroencephalogram (EEG) monitoring and intracarotid single photon emission computed tomography (SPECT) during an intracarotid amobarbital procedure (IAP). METHODS: IAP was performed on 22 patients with temporal lobe epilepsy. CES was defined as the occurrence of significant EEG slowing on the contralateral hemisphere (>50% of the ipsilateral hemisphere slowing) after amobarbital injection. To map the distribution of the amobarbital, we injected a mixture of amobarbital and (99m)technetium ethylcysteinate dimer (99mTc-ECD) into the internal carotid artery and performed a brain SPECT 2 h later. In the SPECT images, regions of interest were determined by ipsilateral and contralateral anterior cerebral artery territories (iACA, cACA), ipsilateral and contralateral middle cerebral artery territories (iMCA, cMCA), and ipsilateral and contralateral posterior cerebral artery territories (iPCA, cPCA), as well as ipsilateral and contralateral anterior and posterior mesial temporal regions (iAMT, cAMT, iPMT, cPMT). The perfusion of amobarbital was interpreted visually in each region. RESULTS: Amobarbital was distributed in the iMCA in all the patients; in the iACA in 20 (90.9%) patients; in the iAMT in 14 (63.5%); and in the iPCA and iPMT in only two (9.1%). CES was observed in 13 (59.1%) patients. Cross-perfusion of amobarbital in limited areas of the cACA were observed in only four of 13 patients. Wada retention memory scores (WRMS) showed no significant difference between the CES- (n = 9) and CES+ (n = 13) groups. CONCLUSIONS: Amobarbital rarely perfused the iPCA territory and the iPMT region and was rarely delivered to the contralateral hemisphere. The occurrence of CES was not related to the cross-perfusion of amobarbital. CES appears to be produced by a transient functional disconnection from the ipsilateral hemisphere. PMID- 10691119 TI - Frequency of bitemporal independent interictal epileptiform discharges in temporal lobe epilepsy. AB - PURPOSE: Bitemporal interictal epileptiform discharges (IEDs) occur in < or =42% of scalp EEGs in patients with temporal lobe epilepsy (TLE) studied with routine EEGs or partial analysis of long-term recordings. METHODS: Twenty-eight patients with TLE demonstrating exclusively unilateral temporal IEDs on routine EEGs underwent 24-h continuous recording. The entire record was visually inspected for epileptiform discharges. We used continuous EEG to assess the significance of long-term recording in detecting bilateral IEDs. RESULTS: Twenty-two patients had left temporal IEDs; 21 had right temporal IEDs. Seventeen (61%) patients had IEDs originating from both the right and left temporal lobes. The probability of detecting bilateral independent IEDs was correlated with the duration of continuous EEG recording. There was no correlation between the number of IEDs originating from one side and the probability of detecting independent IEDs on the other side. The frequencies of IEDs were not correlated with the length of time since onset of epilepsy. CONCLUSIONS: The findings suggest that when long term recordings are performed, the incidence of bilateral discharges in TLE is higher than previously reported in the literature and supports the view that TLE is commonly a bilateral disease. PMID- 10691120 TI - The value of early postictal EEG in children with complex febrile seizures. AB - PURPOSE: To assess the usefulness of an early postictal EEG in neurologically normal children with complex febrile seizures. METHODS: We conducted a retrospective chart review of all neurologically normal children who were hospitalized over a period of 2.5 years after complex febrile seizures, and had an EEG up to 1 week after the seizure. RESULTS: Thirty-three patients (mean age, 17.8 months) qualified for inclusion into the study. Twenty-four patients were qualified as complex cases based on one factor (prolonged in 9, repetitive in 13, and focal in 2). Nine other patients had two complex factors: in six patients, the seizures were long and repetitive; in two patients, the seizures were focal and repetitive; and in one patient, the seizures were long, focal, and repetitive. Thirteen (39%) patients experienced prior febrile seizures. All 33 patients had a normal postictal sleep EEG. Our results indicate with a 95% probability that the true rate of abnormalities in an early postictal EEG performed on otherwise normal children with complex febrile seizures is 8.6% or less. CONCLUSIONS: The yield of abnormalities of an early postictal EEG in this population is low and similar to the reported rate of abnormalities in children with simple febrile seizures. The routine practice of obtaining an early EEG in neurologically normal children with complex febrile seizures is not justified. PMID- 10691121 TI - A multicenter randomized controlled trial on the clinical impact of therapeutic drug monitoring in patients with newly diagnosed epilepsy. The Italian TDM Study Group in Epilepsy. AB - PURPOSE: To assess the clinical impact of monitoring serum concentrations of antiepileptic drugs (AEDs) in patients with newly diagnosed epilepsy. METHODS: One-hundred eighty patients with partial or idiopathic generalized nonabsence epilepsy, aged 6 to 65 years, requiring initiation of treatment with carbamazepine (CBZ), valproate (VPA), phenytoin (PHT), phenobarbital (PB), or primidone (PRM) were randomly allocated to two groups according to an open, prospective parallel-group design. In one group, dosage was adjusted to achieve serum AED concentration within a target range (10-20 microg/ml for PHT, 15-40 microg/ml for PB, 4-11 microg/ml for CBZ, and 40-100 microg/ml for VPA), whereas in the other group, dosage was adjusted on clinical grounds. Patients were followed up for 24 months or until a change in therapeutic strategy was clinically indicated. RESULTS: Baseline characteristics did not differ between the two groups. Most patients with partial epilepsy were treated with CBZ, whereas generalized epilepsies were most commonly managed with PB or VPA. PHT was used only in a small minority of patients. A total of 116 patients completed 2 year follow-up, and there were no differences in exit rate from any cause between the monitored group and the control group. The proportion of assessable patients with mean serum drug levels outside the target range (mostly below range) during the first 6 months of the study was 8% in the monitored group compared with 25% in the control group (p < 0.01). There were no significant differences between the monitored group and the control group with respect to patients achieving 12 month remission (60% vs. 61%), patients remaining seizure free since initiation of treatment (38% vs. 41%), and time to first seizure or 12-month remission. Frequency of adverse effects was almost identical in the two groups. CONCLUSIONS: Only a small minority of patients were treated with PHT, the drug for which serum concentration measurements are most likely to be useful. With the AEDs most commonly used in this study, early implementation of serum AED level monitoring did not improve overall therapeutic outcome. and the majority of patients could be satisfactorily treated by adjusting dose on clinical grounds. Monitoring the serum levels of these drugs in selected patients and in special situations is likely to be more rewarding than routine measurements in a large clinic population. PMID- 10691122 TI - Remote memory in epilepsy. AB - PURPOSE: There is now a considerable amount of research relating to memory functioning in epilepsy. The majority of studies have focused on the retention of new information, and few reports have measured memory for past events. This study aims to redress this and measure the efficiency of remote memory in epilepsy. METHODS: A remote memory questionnaire was prepared and administered to three groups of patients with epilepsy and a control group without epilepsy. The questionnaire assessed knowledge of public events that occurred between 1980 and 1991, inclusive. The epilepsy groups comprised 33 patients with temporal lobe epilepsy (TLE), 33 with extratemporal epilepsy (ExTE), and 10 with primary generalized epilepsy (PGE). Thirty control subjects were tested. RESULTS: Patients with TLE performed significantly less well on the questionnaire than all other groups (p = 0.001), but no effect of laterality was recorded; patients with extratemporal or primary generalised epilepsy did not differ from controls. Performance on the questionnaire was not determined by verbal IQ, educational achievement, social class, or drug treatment, but was related to the number of generalised convulsions that had occurred since 1980. The strongest neuropsychological predictors of performance on this questionnaire were measures of verbal memory. CONCLUSIONS: The study demonstrated weak memory for past events in patients with TLE, thereby providing evidence of a broader memory disturbance in this group than has been previously highlighted. A test of remote memory, such as the one designed for this study, is easy to administer and provides clinically important information not available from conventional neuropsychological tests. PMID- 10691123 TI - Slow-frequency repetitive transcranial magnetic stimulation in a patient with focal cortical dysplasia. AB - PURPOSE: To evaluate the effect of slow-frequency repetitive transcranial magnetic stimulation (SF-rTMS) on interictal epileptiform activity and seizure frequency in a patient with medically refractory partial seizures due to focal cortical dysplasia. METHODS: A 9-cm circular coil was positioned over the area of cortical dysplasia. One hundred stimuli given at 0.5 Hz at 5% below motor threshold were given biweekly for four consecutive weeks. The EEG was recorded for 30 min before and after the first 100 stimuli. The number of seizures during the month of stimulation was compared with that of the month before stimulation. RESULTS: Stimulation was associated with a 70% reduction in the frequency of seizures and a 77% reduction in the frequency of interictal spikes. No seizures occurred during stimulation. CONCLUSIONS: SF-rTMS was safe and well tolerated in this patient. The reduction in seizures and interictal spikes associated with SF rTMS supports the concept of SF-rTMS-induced cortical inhibition. PMID- 10691124 TI - Seizures provoked by blows to the head. AB - The nonepileptic nature of convulsions occurring within seconds of head injury has been commented on. However, we have recently seen a patient in whom relatively trivial blows to the head did seem to give rise to epileptic seizures. PMID- 10691125 TI - Can early postnatal closed head injury induce cortical dysplasia. AB - PURPOSE: Increased availability of surgically resected epileptogenic tissues reveals often unsuspected cortical dysplasia (CD). There is some controversy about the ontogenic stages in which these occur. Although most take place during neuroblast proliferation and migration, there is some evidence for some CD occurring during postmigrational intrinsic cortical organization. It has been shown that various kinds of focal cortical manipulations in rats, if performed within 3-4 postnatal days, lead to the genesis of various cortical malformations including a four-layered microgyrus or an unlayered CD. It is not known whether such events also might occur in the human brain. METHODS: Two children sustained minor head trauma within 4 postnatal days and later developed intractable epilepsy, which was relieved by surgery. Neuropathologic analysis of the resected tissues revealed an unsuspected microdysplastic cortex immediately adjacent to a focal, modest meningeal fibrosis, presumably secondary to the old closed head trauma. RESULTS: The main histologic features were a disorganized, unlayered cortex; abnormal clusters of neurons, often with complex, randomly oriented proximal dendritic patterns with absent apical orientation; the presence of a number of heterotopic small and large neurons in the white matter; absence of inflammatory infiltrates, of hemosiderine, of reactive gliosis, or of an excessive number of blood vessels. The morphologic features in these surgical specimens suggest that these focal malformations occur because of a regional disorder of postmigrational intrinsic cortical remodeling. CONCLUSIONS: The clinical histories and the pathologic findings lend some support to the hypothesis that minor morbid events occuring in the immediate postnatal period may lead to microdysplasia in the human similar to those induced in rat pups. The animal model could be helpful to clarify the genesis of some cases of CD and of the epileptogenicity often manifesting later in life. PMID- 10691126 TI - Hemicranial volume deficits in patients with temporal lobe epilepsy with and without hippocampal sclerosis. PMID- 10691127 TI - Hemicranial volume deficits in patients with temporal lobe epilepsy with and without hippocampal sclerosis. PMID- 10691128 TI - Chromosome of the enterohemorrhagic Escherichia coli O157:H7; comparative analysis with K-12 MG1655 revealed the acquisition of a large amount of foreign DNAs. AB - A complete Xba I and Bln I cleavage map was constructed for the chromosome of an enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain isolated from an outbreak in Sakai City, Japan, in 1996. A comparative chromosome analysis with E. coli K-12 strain MG1655 was made. The EHEC chromosome was approximately 5600 kb in length, 1 Mb larger than that of MG1655. Despite the marked difference in chromosome length, the location and direction of seven rRNA operons of the EHEC strain were similar to those for MG1655. Overall organization of genes common in both strains is also highly conserved. Chromosome expansion was observed throughout the EHEC chromosome, albeit in an uneven manner. A large portion of the chromosome enlargement was observed in the region surrounding the replication terminus, particularly in a segment containing the terA locus. Sample sequencing of 3627 random shotgun clones suggested the presence of approximately 1550 kb strain-specific DNAs on the EHEC chromosome, most of which are likely to be of foreign origin. PMID- 10691129 TI - A large scale structural analysis of cDNAs in a unicellular green alga, Chlamydomonas reinhardtii. I. Generation of 3433 non-redundant expressed sequence tags. AB - To understand genetic information carried in a unicellular green alga, Chlamydomonas reinhardtii, normalized and size-selected cDNA libraries were constructed from cells at photoautotrophic growth, and a total of 11,571 5'-end sequence tags were established. These sequences were grouped into 3433 independent EST species. Similarity search against the public non-redundant protein database indicated that 817 groups showed significant similarity to registered sequences, of which 140 were of previously identified C. reinhardtii genes, but the remaining 2616 species were novel sequences. The coverage of full length protein coding regions was estimated to be over 60%. These cDNA clones and EST sequence information will provide a powerful source for the future genome wide functional analysis of uncharacterized genes. PMID- 10691130 TI - Isolation and mapping of rFUS6, a rice orthologue of Arabidopsis thaliana FUS6. AB - COP9 complex is one of the most important components that act in repressing photomorphogenesis in Arabidopsis thaliana. FUS6 has been identified as one of eight subunits of the COP9 complex in Arabidopsis. Using Arabidopsis Fus6 cDNA as a probe, we screened a rice root cDNA library and a rice genomic library. A 1730 bp cDNA was obtained, which has an open reading frame corresponding to 441-amino acid. This 441 amino acids putative protein has 67% identity with Arabidopsis COP11/FUS6 (AtFUS6) and 40% identity with human GPS1, an AtFUS6 orthologue. So we designated this novel gene as rFUS6. The 6.2-kb genomic sequence of rFUS6 was also obtained. Sequence comparison showed that the rFUS6 gene had six exons and five introns. Sequence inspection of the 5'-flanking region revealed the presence of some potential light-regulated cis-elements such as a G-box, GT-1 binding sites, and a TGACG motif. Southern hybridization with rice total DNA showed that rFUS6 was perhaps a single copy gene. The rFUS6 locus was mapped by hybridization with a rice BAC library membrane and the results showed that rFUS6 had a locus at 16.3 cM of chromosome 1. PMID- 10691131 TI - Structural analysis and complete physical map of Arabidopsis thaliana chromosome 5 including centromeric and telomeric regions. AB - Previously, we have reported a fine physical map of Arabidopsis thaliana chromosome 5, except for the centromeric and telomeric regions, by ordering clones from YAC, P1, TAC, and BAC libraries of the genome consisting of the two contigs of upper arm and lower arm, 11.6 M bases and 14.2 M bases, respectively. Here, the remaining centromeric and telomeric regions of chromosome 5 are completely characterized by the ordering of clones and PCR amplifications. Chromosome 5 of Arabidopsis thaliana ecotype Columbia is about 28.4 M bases long. The centromeric region is estimated at about 2 M bases long between two 5S-rDNA clusters. The 180-bp repeat region mainly consists of blocks of 180-bp tandem family and various type retroelements dispersed over a 500-kb region. The telomeric regions of chromosome 5 are characterized by PCR cloning, sequencing and hybridization. The telomere repeats at both ends are about 2.5-kb long and interestingly, telomere-associated repeats (approximately 700 bp) are found near both ends of chromosome 5. PMID- 10691132 TI - Sequence analysis of a total of three megabases of DNA in two regions of chromosome 8p. AB - Large-scale sequencing of genomic regions and in silico gene trapping together represent a highly efficient and powerful approach for identifying novel genes. We performed megabase-level sequence analyses of two genomic regions on human chromosome 8p (8p11.2 and 8p22-->p21.3), after covering those segments with sequence-ready contigs composed of 74 cosmids, 14 BACs, and three PAC clones. We determined continuous nucleotide sequences of 1,856,753 bases on 8p11.2 and 1,210,381 bases on 8p22-->p21.3 by combining the shotgun and primer-walking methods. In silico gene trapping identified four novel genes in the 8p11.2 region and, in the 8p22-->p21.3 region, six known genes (PRLTS, PCM1, MTAMR7, HCAT2, HFREP-1 and PHP) and three novel genes. The distribution of Alu and LINE1 repetitive elements and the densities of predicted exons were different in each region, and Alu-rich portions contained more exonic sequences than LINE1-rich areas. PMID- 10691133 TI - Sequence-ready 1-Mb YAC, BAC and cosmid contigs covering the distal imprinted region of mouse chromosome 7. AB - We have constructed approximately 1-Mb contigs of yeast artificial chromosome (YAC), bacterial artificial chromosome (BAC) and cosmid clones covering the imprinted region in mouse chromosome band 7F4/F5. This region is syntenic to human chromosome 11p15.5, which is associated with Beckwith-Wiedemann syndrome (BWS) and certain childhood and adult tumors. These contigs provide the basis for genomic sequencing, identification of genes and their regulatory elements, and functional studies in transgenic and knockout mice, which should be of help to understand not only the mechanisms of imprinting but also the molecular events involved in the genesis of BWS and tumors. PMID- 10691134 TI - A binary vector plasmid for gene expression in plant cells that is stably maintained in Agrobacterium cells. PMID- 10691135 TI - Computer-assisted three-dimensional surgical planning and simulation: 3D color facial model generation. AB - A scheme for texture mapping a 3D individualized color photo-realistic facial model from real color portraits and CT data is described. First, 3D CT images including both soft and hard tissues should be reconstructed from sequential CT slices, using a surface rendering technique. Facial features are extracted from 3D soft tissue. A generic mesh is individualized by correspondence matching and interpolation from those feature vertices. Three digitized color portraits with the "third" dimension from reconstructed soft tissue are blended and texture mapped onto the 3D head model (mesh). A color simulated human head generated from frontal, right and left real color portraits can be viewed from an arbitrary angle in an inexpensive and user-friendly conventional personal computer. This scheme is the basic procedure in 3D computer-assisted simulation surgery. PMID- 10691137 TI - Laser scanning of the ear identifying the shape and position in subjects with normal facial symmetry. AB - The objective of the present study was to discover if dimensional measurements of the ear could be determined with a laser scanning technique and whether or not the location of landmarks of the ear could be reliably measured with respect to those on the midline of the face. Computer-generated images were created from laser scans of 20 subjects. Dimensional measurements were made between landmarks on the ear and face. Differences between repeated dimensional measurements of the ear were very small, as were those measurements made between landmarks on the ear to the midline of the face. Differences between dimensions of the left and right ears were observed, but were of a small magnitude. The results suggest that the dimensions of the ear and its position with respect to landmarks in the midline of the face can be reliably measured on normal subjects and that laser scanning is a useful technique for planning and monitoring facial reconstruction of the ear. PMID- 10691136 TI - Computer-assisted three-dimensional surgical planning and simulation: 3D virtual osteotomy. AB - A computer-assisted three-dimensional virtual osteotomy system for orthognathic surgery (CAVOS) is presented. The virtual reality workbench is used for surgical planning. The surgeon immerses in a virtual reality environment with stereo eyewear, holds a virtual "scalpel" (3D Mouse) and operates on a "real" patient (3D visualization) to obtain pre-surgical prediction (3D bony segment movements). Virtual surgery on a computer-generated 3D head model is simulated and can be visualized from any arbitrary viewing point in a personal computer system. PMID- 10691138 TI - Three-layer closure of an oroantral-cutaneous defect. AB - Reconstruction of oroantral defects, which are usually caused by tumor resection, is challenging. These defects become an even more difficult problem when they comprise multiple layers including oral mucosa, subcutaneous tissue, muscle and skin. This paper describes such a case in which a three-layer closure using a palatal flap, a buccal fat pad flap and a local skin flap was successfully performed. PMID- 10691139 TI - Tooth-borne distraction to widen the mandible. Technical note. AB - Based on reported animal studies, the case report presented demonstrates the use of tooth-borne distraction osteogenesis for mandibular widening of 1 cm in order to avoid extraoral distraction devices. Detrimental side effects on healthy anchoring teeth are not likely. PMID- 10691140 TI - Traumatic optic neuropathy. A case report. AB - A case of visual loss following cranio-maxillofacial trauma is reported. The patient had a sudden partial blindness associated with a fracture of the roof, medial and lateral orbital walls. Access to the orbit was achieved through a transethmoidal approach using the Howarth-Lynch medial incision and resecting the bone fragments which impinged on the optic nerve. The patient had total return of visual acuity, without surgical complications. The role of orbital and optic decompression in the management of patients with traumatic optic neuropathy is discussed. Its indications are controversial and the procedure should be considered only within the context of the specific needs of the individual patient. PMID- 10691141 TI - The role of vital tissue staining in the marginal control of oral squamous cell carcinoma. AB - Since vital staining has been advocated as a sensitive method of displaying epithelial atypia, 14 oral squamous cell carcinomas were stained immediately preoperatively with toluidine blue and then resected 1 cm outside clinically abnormal or positively staining tissue. The integrity of the entire tissue margin was histologically examined in each case. Whilst vital staining delineated all 14 invasive carcinomas at the centre of each resected specimen, 10 foci of carcinoma in-situ or severe dysplasia were identified not to have stained at the resection margins. Toluidine blue may, therefore, be an adjunct in identifying invasive tumour at mucosal resection margins. However, it would appear to be of no benefit in delineating positive resection margins due to carcinoma-in-situ or severe dysplasia, and hence it may be of little value in reducing the incidence of local recurrences. PMID- 10691142 TI - Nuclear DNA content, an adjunct to p53 and Ki-67 as a marker of resistance to radiation therapy in oral cavity and pharyngeal squamous cell carcinoma. AB - Factors of prognosis and radioresistance in oral cavity and pharyngeal squamous cell carcinoma (OCPSCC) are limited. In the present study, the usefulness of tumor DNA content in predicting radioresistance in patients with OCPSCC has been investigated. Radioresistance has been defined as local recurrence or tumor persistence after radiation therapy. DNA-ploidy analysis was performed by static cytometry on smears of cell suspensions obtained from formalin-fixed paraffin embedded material and stained with Feulgen. DNA-ploidy was correlated with the proliferation rate (Ki-67) and p53 protein accumulation obtained by immunohistochemistry. The follow-up of patients ranged from 8 to 62 months. Radioresistance was more common in non-diploid tumors; 14/28 (50%) non-diploid tumors recurred, whereas only 3 (10.7%) out of 28 diploid tumors had local failure (P=0.0019). Proliferation rate and p53 accumulation, evaluated by immunohistochemistry, also added prognostic information. Twelve out of 14 failures were from non-diploid tumors with a low proliferation rate (Ki-67<20%), whereas none of 20 p53-negative diploid tumors developed recurrences. This study showed that non-diploid tumors responded poorly to radiotherapy. DNA content appeared, therefore, as a significant prognostic marker for the evaluation of OCPSCC in patients receiving radiation therapy. This study also showed that DNA content adds information to p53 accumulation and the proliferation rate (Ki-67) for the purposes of determining patient management. PMID- 10691143 TI - Chromatic analysis of autofluorescence emitted from squamous cell carcinomas arising in the oral cavity: a preliminary study. AB - Chromatic analysis was carried out to characterize the color quality of autofluorescence emitted from oral squamous cell carcinomas (SCC) and to objectively compare autofluorescence among various tissues. The following specimens were studied: 33 SCC, 3 epithelial dysplasias, 39 benign lesions, 31 dorsa of the tongue and 18 dental plaques. Autofluorescence depicted on fluorescence photographs was measured with a chroma meter. Chromatic values of autofluorescence differed significantly between SCC and non-cancerous tissues and between different stages of SCC. Autofluorescence of SCC tended to shift from orange to reddish orange with advancement of stage. These results suggest that autofluorescence of oral SCC chromatically differs from that of other tissues and depends on the stage of cancer. PMID- 10691144 TI - Traumatic cervical cystic hygroma. AB - An unusual case of traumatic cervical cystic lymphangioma in an adult man is presented. PMID- 10691145 TI - Diffuse chronic sclerosing osteomyelitis and the synovitis, acne, pustolosis, hyperostosis, osteitis (SAPHO) syndrome in two sisters. AB - Two sisters with diffuse chronic sclerosing osteomyelitis of the mandible and the humerus and the synovitis, acne, pustolosis, hyperostosis and osteitis syndrome (SAPHO syndrome) are presented. The diagnoses of diffuse chronic sclerosing osteomyelitis at the age of 12 years and 27 years, respectively, were based on typical medical history, clinical symptoms and radiographic, histologic and scintigraphic findings. Because skin lesions and scintigraphic enhancement of the sternoclavicular joints with hyperostosis were present, a SAPHO syndrome was diagnosed in both sisters. Microbiological cultures of biopsy specimens revealed coagulase-negative Staphylococcus aureus at the humerus and Haemophilus parainfluenzae, Streptococcus, Actinomyces and Veilonella species at the mandible. Repeated operative procedures, including decortications, resection and reconstruction, and multiple histologic and microbiologic studies were performed over a period of up to 20 years. Since HLA typing yielded identical gene loci, we suggest that hereditary and autoimmune factors may play a role in the pathogenesis of these cases. PMID- 10691146 TI - Biomechanical and histological studies of particulate hydroxylapatite implanted in femur bone defects of adult dogs. AB - The purpose of this study was to investigate the biomechanical behavior and the histology of particulate hydroxylapatite (HA) implanted in bone defects created in femurs of 48 adult mongrel dogs. The bone defects, measuring 10 x 10 x 25 mm with or without HA implantation, were allowed to heal until the end of the 1st, 2nd, 3rd, 6th, 9th and 12th months after surgery. The harvested tissue specimens from the implant sites were processed into long cubic or rectangular prismatic forms. Their mechanical strengths were assessed using compression and shearing tests to measure the maximal compressive and shear stress by force loading on the middle portions of the specimens. Histological sections of each stage were processed with both decalcified and nondecalcified methods. The results showed that the mechanical strength of the tested specimens was closely related to the tissue regeneration within the bone defect. Progressive regeneration of new bone was observed at each stage, with complete bone formation at the 9-month period. A consistent increase in both maximal compressive stress and maximal shear stress was noted at each stage, responsive to the ossification and maturation of the regenerating tissue within the bone defect. This animal model provides a novel approach to directly estimate the tissue strength of the HA-implanted bone defect. PMID- 10691147 TI - Osteoinduction by recombinant human bone morphogenetic protein-2 at intramuscular, intermuscular, subcutaneous and intrafatty sites. AB - To compare the osteoinductive activity of recombinant human bone morphogenetic protein-2 (rhBMP-2) at various sites in rats, 5 microg of rhBMP-2 were implanted into various sites, using atelopeptide type-I collagen (CL) as a carrier (BMP groups). CL implantation was used as a control. Forty Wistar rats were divided into intramuscular, intermuscular, subcutaneous and intrafatty site groups (IrM, IeM, SC and IF, respectively). Bone formation was evaluated radiographically, histologically and biochemically 21 days after implantation. In the BMP groups, the alkaline phosphatase activity and calcium content at all sites were higher than those in the control groups. Among the BMP groups, the new bone formation was highest in the IrM and lowest in the IF radiographically, histologically and biochemically. Five microg of rhBMP-2, a relatively low dose, induced adequate new bone formation in all sites. The variations of osteoinductive activity of rhBMP-2 in various sites may be due to differences in the blood supply. PMID- 10691148 TI - An image-based approach for designing and manufacturing craniofacial scaffolds. AB - Bone tissue engineering (BTE), which combines biomaterial scaffolds with biologically active factors, holds tremendous promise for reconstructing craniofacial defects. A significant challenge in craniofacial reconstructive BTE applications is the complex patient-specific geometry that must be reconstructed. In this paper, we present an image-based approach for designing and manufacturing patient-specific craniofacial biomaterial scaffolds directly from CT or MRI data. In this approach, voxel density distribution is used to define scaffold topology. The scaffold design topology is created using image processing techniques. This voxel density distribution is then converted to data that can be used to drive a Solid Free-Form Fabrication machine to either directly build the scaffold or build a mold for the scaffold. Several preliminary applications for craniofacial surgery, including a mandibular condyle scaffold, an orbital floor scaffold, and a general mandibular defect scaffold, are illustrated. Finally, we show applications to in vivo models. PMID- 10691149 TI - Unusual periosteal reaction caused by an accidentally displaced dental root. PMID- 10691150 TI - The effect of suction drainage on microvascular anastomosis. PMID- 10691151 TI - Standardization of HbA1c measurement--the issues. PMID- 10691152 TI - Standardization of HbA1c measurements--a consensus statement. PMID- 10691153 TI - Freezing method affects the concentration and variability of urine proteins and the interpretation of data on microalbuminuria. The Oxford Regional Prospective Study Group. AB - AIMS: Microalbuminuria and, to a lesser extent, renal tubular proteins are widely used in the early detection of incipient nephropathy in diabetes mellitus. Recent reports have indicated detrimental effects of storage at -20 degrees C on urine proteins. This study investigated the effects of storage on the measurement of urine proteins and discusses implications for the interpretation of data. METHODS: Two-hundred and sixty-eight specimens, collected from children with Type 1 diabetes, split into duplicate aliquots and stored at -20 degrees C and -70 degrees C, respectively, for 6-8 months, were analysed for albumin, retinol binding protein, N-acetyl glucosaminidase and creatinine, in the same assays to eliminate inter-assay variability. Two independent non-diabetic cohorts of children provided urine specimens, which were stored at -20 degrees C for one cohort and -70 degrees C for the other, to determine normal ranges for urine proteins. RESULTS: Storage at -20 degrees C led to a variable underestimation of all three urine proteins in 20% of specimens. Creatinine was unaffected. This underestimation was greater in more concentrated urine (r2 = 0.38, P < 0.001, n = 262). Consequently storage at -20 degrees C increased the variance of the albumin/ creatinine ratio more than the variance of albumin concentration. Temperature of storage affected the normal range, which was 0.1-2.1 mg/mmol at 20 degrees C compared to 0.3-3.1 mg/mmol at -70 degrees C. The prevalence of microalbuminuria (> 2SD above the geometric mean in non-diabetic specimens stored at -20 degrees C) was 27% after storage at -70 degrees C vs. 24% after -20 degrees C. The prevalence of microalbuminuria (>2SD above the geometric mean in nondiabetic specimens stored at -70 degrees C) was 21% after storage at -70 degrees C vs. 17% after -20 degrees C. CONCLUSIONS: Urine proteins are significantly but variably underestimated after storage at -20 degrees C. These effects account for increased variance and differences in the normal range, but have less effect on the detection of microalbuminuria than might be predicted. PMID- 10691154 TI - Predicting the occurrence of diabetes mellitus in recipients of heart transplants. AB - AIMS: To establish the incidence of post-transplant diabetes mellitus (PTDM) and factors predictive of its development. METHODS: This was a retrospective review (using hospital records and transplant database) of 97 consecutive adult patients who underwent cardiac transplantation at St Vincent's Hospital, Sydney, Australia. RESULTS: Mean follow-up was 27 months. Excluding five patients who had pre-existing diabetes, the cumulative incidence of PTDM was 15.7%. Pre-transplant random blood glucose (5.6 +/- 0.8 vs. 5.2 +/- 0.6 mmol/l, P<0.05), family history (46% vs. 15%, P<0.05) and a continuing requirement for insulin on the second post transplant day (54% vs. 15%, P< 0.01) differed in those who developed PTDM as opposed to those who remained free of diabetes. Patients who developed PTDM had received slightly higher mean doses of prednisolone at three months (0.21 +/- 0.03 vs. 0.19 +/- 0.03 mg. kg(-1)/day(-1), P<0.01). Of the factors identifiable prior to initial hospital discharge, only family history of diabetes mellitus and second post-transplant day insulin requirement independently predicted the occurrence of PTDM. CONCLUSIONS: A family history of diabetes and the need for insulin beyond the first 24 h after transplantation are factors identifiable prior to hospital discharge, which predict patients at risk of developing PTDM. In such patients, consideration to minimizing the dose of glucocorticoids should be given where possible. PMID- 10691155 TI - Acarbose treatment does not change the habitual diet of patients with Type 2 diabetes mellitus. The Finnish Acargbos Study Group. AB - AIMS: It has been speculated that acarbose treatment in patients with Type 2 diabetes mellitus might induce changes in diet as a result of its adverse gastrointestinal effects. The aim of this study was to determine whether poor metabolic control can be improved by acarbose, and whether this might be because the acarbose supplementation provokes changes in diet. METHODS: Poorly controlled Type 2 diabetic patients treated with oral hypoglycaemic agents (OHA) were randomized into either acarbose (100 mg t.d.s.) or placebo treatment. The double blind treatment lasted for 24 weeks. Four-day food diaries and blood samples for efficacy analysis were collected at 0, 4, 12, and 24 weeks. Thirty-six acarbose and 39 placebo-treated patients completed the trial and were included in the final analyses. RESULTS: At 24 weeks the baseline adjusted means of fasting, 1 and 2-h postprandial blood glucose values were 9.3 vs. 10.5 (P=0.02), 11.6 vs. 14.5 (P<0.001) and 11.0 vs. 13.7 mmol/l (P<0.001) and HbA1 9.3% vs. 10.2% (P=0.002) in the acarbose and placebo groups, respectively. No significant differences in nutrient intakes between groups were observed. The energy intake and energy proportion of fat and carbohydrates remained unchanged in both groups. CONCLUSIONS: Acarbose significantly improves metabolic control in Type 2 diabetic patients poorly controlled with oral hypoglycaemic agents. This effect seems not to be a result of concomitant involuntary dietary changes, since acarbose did not induce modifications in diet during the study. PMID- 10691156 TI - Universal vs. risk factor-based screening for gestational diabetes mellitus: detection rates, gestation at diagnosis and outcome. AB - AIMS: Gestational diabetes mellitus (GDM) is associated with adverse maternal and fetal outcome. Screening for GDM is therefore recommended but the best screening method remains controversial. This prospective, randomized study compared a risk factor-based screening programme with a universally based one. METHODS: Subjects were randomized at booking to one of two groups: the risk factor group had a 3-h 100-g oral glucose tolerance test (OGTT) at 32 weeks if any risk factor for GDM was present; the universal group had a 50-g glucose challenge test performed and if their plasma glucose at 1 h was > or = 7.8 mmol/l, a formal 3-h 100-g OGTT was then performed. RESULTS: Universal screening detected a prevalence of GDM of 2.7%, significantly more than the 1.45% detected in the risk factor screened group (P<0.03). Universal screening facilitated earlier diagnosis than risk factor screening - mean gestation 30 +/- 2.6 weeks vs. 33 +/- 3.7 weeks (P<0.05). A higher rate of spontaneous vaginal delivery at term, and lower rates of macrosomia, Caesarean section, prematurity, pre-eclampsia and admission to neonatal intensive care unit were observed in the universally screened, early diagnosis group. CONCLUSIONS: Universal screening for GDM is superior to risk factor based screening-detecting more cases, facilitating early diagnosis and is associated with improved pregnancy outcome. PMID- 10691157 TI - Perinatal mortality in Type 2 diabetes mellitus. AB - AIMS: In many parts of the world the number of pregnancies in women with Type 2 diabetes mellitus (DM) now exceeds that in women with Type 1 DM, but there are few data published on perinatal mortality in Type 2 DM. This study reports observational data on perinatal mortality in Type 2 DM from a population with a high background rate of this disorder. METHODS: Over a 12-year period (1985-1997) at the Diabetes Clinic at National Women's Hospital, Auckland, there were 434 pregnancies in women with Type 2 DM (256 known and 178 diagnosed with gestational diabetes mellitus (GDM), but confirmed to have Type 2 DM early post-partum), 160 pregnancies in women with Type 1 DM and 932 in women with GDM. Perinatal mortality was classified as either intermediate fetal death (20-28 weeks' gestation), late fetal death (28 weeks' gestation to term) or early neonatal death (up to 1 month post-partum). RESULTS: The perinatal mortality in Type 2DM was 46.1/1,000, significantly higher than the rates for the general population (12.5), Type 1 DM (12.5) and GDM (8.9) (P < 0.0001). Congenital malformations accounted for only 10% of the perinatal mortality. There was a seven-fold increase in the rate of late fetal death and 2.5-fold increase in the rates of intermediate fetal and late neonatal death. Subjects with Type 2 DM were significantly older and more obese than subjects with Type 1 DM, and presented later to the diabetes service. CONCLUSIONS: Perinatal mortality in Type 2 DM is significantly increased, mainly owing to an excess of late fetal death. Maternal factors such as obesity may be important contributors to the high perinatal mortality. Women diagnosed with GDM who have unrecognized Type 2 DM are also at high risk, but perinatal mortality is low in women with milder degrees of glucose intolerance in pregnancy. PMID- 10691158 TI - Addition of low-dose rosiglitazone to sulphonylurea therapy improves glycaemic control in Type 2 diabetic patients. AB - AIMS: This study was designed to test the efficacy and safety of low-dose rosiglitazone, a potent, insulin-sensitizing thiazolidinedione, in combination with sulphonylurea in Type 2 diabetic patients. METHODS: For the intention-to treat analysis, 574 patients (59% male, mean age 61 years) were available, randomized to receive 26 weeks of twice-daily placebo (n = 192), rosiglitazone 1 mg (n = 199) or rosiglitazone 2 mg (n = 183) in addition to existing sulphonylurea treatment with gliclazide (47.6% of patients), glibenclamide (41.8%) or glipizide (9.4%) (two patients were taking carbutamide or glimepiride). Change in haemoglobin A1c (HbA1c), fasting plasma glucose (FPG), fructosamine, insulin, C-peptide, albumin, and lipids were measured, and safety was evaluated. RESULTS: Mean baseline HbA1c was 9.2% and FPG was 11.4 mmol/l. Rosiglitazone at doses of 1 and 2 mg b.d. plus sulphonylurea produced significant decreases, compared with sulphonylurea plus placebo, in HbA1c (-0.59% and -1.03%, respectively; both P<0.0001) and FPG (1.35 mmol/l and 2.44 mmol/l, respectively; both P<0.0001). Both HDL-cholesterol and LDL-cholesterol increased and potentially beneficial decreases in non-esterified fatty acids and gamma glutamyl transpeptidase levels were seen in both rosiglitazone groups. The overall incidence of adverse experiences was similar in all three treatment groups, with no significant cardiac events, hypoglycaemia or hepatotoxicity. CONCLUSIONS: Overall, the combination of rosiglitazone and a sulphonylurea was safe, well tolerated and effective in patients with Type 2 diabetes. PMID- 10691159 TI - Placental size and large-for-gestational-age infants in women with abnormal glucose tolerance in pregnancy. AB - AIMS: To determine if the placental size is disproportionately increased in the large-for-gestational age infants in pregnancies complicated by impaired glucose tolerance. METHODS: A retrospective study was performed on 568 consecutive singleton pregnancies complicated by gestational impaired glucose tolerance controlled with diet and who delivered within a 15-month period. The cases were categorized by the infant birthweight percentile into three groups, i.e. small for-gestational age (< 10th percentile), appropriate-for-gestational age (10th to 90th percentile) and large-for-gestational age (> 90th percentile). Maternal and infant anthropometric data, glycaemic status, and placental weight-to-birthweight ratio were compared among the three groups. RESULTS: The infant body mass index and placental weight showed a significantly increasing trend from the small-for gestational age to the large-for-gestational age groups, but there was no significant difference in the placental weight-to-birthweight ratio, values of the oral glucose tolerance test, or haemoglobin A1c among the three groups. On the other hand, the maternal body mass index before pregnancy and at delivery were significantly higher in the large-for-gestational age group. The placental weight, but not the ratio, was significantly correlated with the maternal body mass index before pregnancy and at delivery (P < 0.001). CONCLUSIONS: The results indicate that the placenta is not disproportionately bigger, and therefore unlikely to be the cause, in large-for-gestational age infants. Maternal size appeared to be the major determinant of birthweight percentile ranking in pregnancies with gestational impaired glucose tolerance. PMID- 10691160 TI - Leisure-time physical activity at weekends and the risk of Type 2 diabetes mellitus in Japanese men: the Osaka Health Survey. AB - AIMS: To investigate association between leisure-time physical activity at weekends and the risk of developing Type 2 diabetes mellitus (DM). METHODS: Prospective examination of 6,013 Japanese men aged 35-60 years who were free of DM, impaired fasting glycaemia, or hypertension at study entry. Type 2 DM was defined by a fasting plasma glucose level > or = 7.0 mmol/l or a 2-h post-load plasma glucose level > or =11.1 mmol/l. Data on physical activity obtained from questionnaires consisted of overall leisure-time physical activity weekly and leisure-time physical activity at weekends. RESULTS: During the 59,966 person years follow-up, 444 cases developed Type 2 DM. Regular physical exercise at least once a week was associated with a reduced risk of Type 2 DM. After adjustments for age, body mass index, daily alcohol consumption, smoking habits, blood pressure levels and a parental history of Type 2 DM, men who engaged in regular physical exercise at least once a week had a relative risk of Type 2 DM of 0.75 (95% CI, 0.61-0.93) compared with men engaging in exercise less often. Even vigorous activity only once a week at weekends was associated with a reduced risk of Type 2 DM. Men who engaged in vigorous activity at least once a week at weekends had a multiple-adjusted relative risk of Type 2 DM of 0.55 (95% CI, 0.35 0.88) compared with sedentary men. CONCLUSIONS: Regular physical exercise at least once a week and vigorous activity even only once a week at weekends are associated with a decreased risk of Type 2 DM. PMID- 10691161 TI - Incidence of Type 1 diabetes mellitus in children aged 0-14 in Japan, 1986-1990, including an analysis for seasonality of onset and month of birth: JDS study. The Data Committee for Childhood Diabetes of the Japan Diabetes Society (JDS). AB - AIMS: To detect the incidence of childhood Type 1 diabetes mellitus (DM) (0-14 years) in Japan and to find out whether there is a seasonal pattern in the onset of disease and month of birth of children with diabetes. METHODS: Ascertained data for the period 1986-1990 could be collected in 35 out of 47 local government areas representing 69.4% of the childhood population (aged 0-14 years) of Japan. RESULTS: A total of 1,260 children with Type 1 DM were identified (738 girls, 522 boys). With age there was a progressive increase in incidence from 0.7 to 2.1/10(5) in boys and from 0.6 to 3.5/10(5) in girls. With the exception of 1987, when a coxsackie B3 virus epidemic was registered, no seasonal variation in the month of onset was observed, nor was a seasonal pattern of the month of birth registered in this cohort. CONCLUSIONS: Compared to European countries, the USA and Israel, the Japanese cohort of children with diabetes presents the following differences: the incidence is much lower, there is a preponderance of girls and there is (with one exception) no seasonal pattern. PMID- 10691162 TI - Type 1 diabetes mellitus in Czech children diagnosed in 1990-1997: a significant increase in incidence and male predominance in the age group 0-4 years. Collaborators of the Czech Childhood Diabetes Registry. AB - AIMS: To overview total, age-and sex-specific incidence rates of type 1 diabetes mellitus and their trends in Czech children 0-14 years of age in the period of 1990-1997. METHODS: Type 1 DM cases were ascertained by two independent sources, data of general population were obtained from the annual demographic reports of the State Statistic Bureau. Incidence rates were computed using both ascertainment sources combined. RESULTS: In the study period 1.1.1990-31.12.1997, the total incidence was 10.1 (95% CI 9.6-10.6) per 100,000/year in both sexes, 10.0 (95% CI 9.4-10.7) in boys, and 10.2 (95% CI 9.5-11.0) in girls. The total age-standardized incidence was 9.9 (95% CI 9.4-10.4). The total incidence had a significant increasing trend over the study period (P= 10(-4), annual increment 4.3%). A significant increasing trend was also found in the groups of children 0 4 (P = 0.033, increment 6.9%) and 5-9 years at diagnosis (P = 0.038, increment 4.8%). Statistically significant male predominance was observed in the group diagnosed at age 0-4 years (boys/girls ratio of incidence 1.33, P = 0.035). CONCLUSIONS: We report the first population-based epidemiological data on incidence of childhood Type 1 DM in the Czech Republic. The incidence has increased significantly during the last 8 years. The present incidence is at an intermediate level compared to other European countries. PMID- 10691163 TI - The care of students with insulin-treated diabetes mellitus living in university accommodation: scope for improvement? AB - Concern has been expressed about the welfare of young adults with Type 1 diabetes mellitus who leave home to attend university or college for tertiary education. This has been highlighted by the local experience in Edinburgh of two male students with Type 1 diabetes, both of whom died from metabolic complications of diabetes during their first term at universities distant from their homes. One student died following the development of cerebral oedema secondary to diabetic ketoacidosis, which was probably precipitated by prolonged coma after an episode of severe hypoglycaemia. Another student, who was found 'dead in bed', had a history of previous severe hypoglycaemia. At a Fatal Accident Inquiry in Edinburgh, held following the death of the first student, recommendations were made to improve the care and personal safety of students with diabetes living in university accommodation. Despite the report being circulated to all Scottish universities, the second student died within three years of the inquiry. Further efforts to protect the welfare of students with Type 1 diabetes who are attending centres for tertiary education away from their home environment may require the more active participation by diabetes healthcare professionals. PMID- 10691164 TI - 'Sausage toe': a reliable sign of underlying osteomyelitis. AB - AIMS: To follow-up patients with a 'sausage' deformity of the toe associated with local neuropathic ulceration to confirm the diagnosis of underlying osteomyelitis. This was based on our observation that some diabetic patients with suspected pedal osteomyelitis with a local neuropathic ulcer have a 'sausage' deformity of a toe. METHODS: Over a period of 2 years, 14 patients with foot ulcers, who were observed to have the 'sausage' deformity of a toe in the diabetic foot clinic were followed up and investigated. RESULTS: Underlying osteomyelitis was confirmed in six on the very first X-ray examination. A further seven had osteomyelitis diagnosed on bone scanning. Both the X-ray and the bone scan were equivocal in one patient, whose ulcer only healed after an 8-week course of antibiotics. Antibiotic therapy was successful in 11 patients and three patients required amputation of the affected toe. Following successful treatment, there was full resolution of the 'sausage toe' in the majority. CONCLUSIONS: The appearance of a 'sausage toe' should alert the physician of the possibility of underlying osteomyelitis in diabetic foot, so that prompt treatment can be commenced with antibiotics. PMID- 10691165 TI - Adult learning and continuing education. PMID- 10691166 TI - Persistent vomiting in patients with diabetes. PMID- 10691167 TI - Metformin: a useful adjunct to insulin therapy? PMID- 10691168 TI - Type 1 diabetes mellitus and hepatic sinusoidal fibrosis. PMID- 10691169 TI - Electrocardiographic findings in a middle-aged African population in the Seychelles islands. AB - This study describes major electrocardiogram (ECG) measurements and diagnoses in a population of African individuals; most reference data have been collected in Caucasian populations and evidence exists for interethnic differences in ECG findings. This study was conducted in the Seychelles islands (Indian Ocean) and included 709 black individuals (343 men and 366 women) aged 25 to 64 years randomly selected from the general population. Resting ECG were recorded by using a validated ECG unit equipped with a measurement and interpretation software (Cardiovit AT-6, Schiller, Switzerland). The epidemiology of 14 basic ECG measurements, 6 composite criteria for left ventricular hypertrophy and 19 specific ECG diagnoses including abnormal rhythms, conduction abnormalities, repolarization abnormalities, and myocardial infarction were examined. Substantial gender and age differences were found for several ECG parameters. Moreover, tracings recorded in African individuals of the Seychelles differed from those collected similarly in Caucasian populations in many respects. For instance, heart rate was approximately 5 beats per minute lower in the African individuals than in selected Caucasian populations, prevalence of first degree atrio-ventricular block was especially high (4.8%), and the average Sokolow-Lyon voltage was markedly higher in African individuals of the Seychelles compared with black and white Americans. The integrated interpretation software detected "old myocardial infarction" in 3.8% of men and 0% of women and "old myocardial infarction possible" in 6.1% and 3%, respectively. Cardiac infarction injury scores are also provided. In conclusion, the study provides reference values for ECG findings in a specific population of people of African descent and stresses the need to systematically consider gender, age, and ethnicity when interpreting ECG tracings in individuals. PMID- 10691170 TI - Influence of autonomic tone on the filtered QRS duration from signal averaged electrocardiograms in healthy volunteers. AB - We recently reported that signal averaged electrocardiograms (SAECG) measurements possess a circadian rhythm and are closely related to heart rate or heart rate variability in healthy volunteers. This study determines the influence of autonomic tone on the filtered QRS (f-QRS) duration from SAECG by using pharmacologic autonomic blockade and exercise in healthy volunteers. Eleven healthy male volunteers were studied. Three protocols were designed to study the effects of exercise (Ex) under control conditions, beta adrenergic blockade or double blockade. SAECGs and heart rate variability (LF and HF: low and high frequency power, LF/HF ratio) were determined from Holter recordings. Ex significantly decreased the f-QRS duration and HF and significantly increased heart rate and LF/HF. Ex during beta adrenergic blockade significantly increased heart rate and decreased f-QRS duration and HF, but did not change LF/HF. Ex during double blockade did not affect the f-QRS duration, HF, or LF/HF. The changes in f-QRS duration induced during Ex, autonomic blockade, or both were inversely correlated with changes in heart rate and LF/HF and positively correlated with changes in HF. These data suggest that f-QRS duration in healthy subjects is shortened by Ex with increased sympathetic tone or decreased parasympathetic tone or the combination. PMID- 10691171 TI - Positional changes of spatial QRS- and ST-segment variables in normal subjects: implications for continuous vectorcardiography monitoring during myocardial ischemia. AB - Electrocardiographic QRS- and ST-segment changes are to be expected during changes in body posture. We prospectively analyzed the influence of changes in body position on continuous vectorcardiography monitoring of QRS-vector difference (QRS-VD) and ST change-vector magnitude (STC-VM) according to the currently used criteria of myocardial ischemia in 21 normal subjects. Fifteen (71%) and 6 (29%) subjects had significant positional QRS-VD and STC-VM changes, respectively. Vectorcardiography changes were most frequent and pronounced in the left lateral position. An alternative to the existing criterion of ischemia is proposed to improve the specificity of STC-VM. Subjects with positional QRS-VD changes had higher mean STC-VM values as compared with those without such changes. Otherwise no characteristics among those with positional vectorcardiography changes could be identified. There was no statistically significant association between positional QRS-VD and STC-VM changes (R = .13, P = .57). We conclude that the clinical use of QRS-VD in its present form for continuous vectorcardiography monitoring of myocardial ischemia seems to be of limited practical value, because of the presence of frequent "pseudo-ischemic" changes. STC-VM seems to have a significant potential of continuous vectorcardiography monitoring. However, an indicator of body position change or even an algorithm enabling on-line correction for positional vectorcardiography changes seems to be essential to improve the accuracy of this technique in identifying myocardial ischemia. PMID- 10691172 TI - Age-related changes in the magnitude of ventricular depolarization vector: analyses by magnetocardiogram. AB - The magnetocardiogram has the beneficial feature that permits the strength and location of the current dipole to be estimated. This study examines the issue of whether the magnitude of the heart current during depolarization phase was influenced by the age of healthy subjects. The magnetocardiograms were recorded by means of a second-derivative SQUID (superconducting quantum interference device) magnetometer (BT Corp, Model BMP, San Diego, CA) in 150 healthy subjects. The subjects were subgrouped into 5 age-based categories according to the age. The current dipole of the maximum QRS complex was determined from isofield contour maps during the ventricular depolarization phase, and no significant differences were observed in the magnitude in the current source for any age category. However, the amplitudes of the RV5 and SV1 + RV5 in the standard electrocardiogram were larger in 65 to 74-year-old women than other age groups, and the SV1 + RV5 was smaller for the 45 to 74-year-old men than for the men aged 25 to 44 years. These findings suggest that the age-associated changes in the QRS complex observed by the electrocardiogram are caused by increased electric resistance and not by the heart current itself. The results additionally suggest that no effects of aging were observed in the actual heart current of the heart during the depolarization phase. PMID- 10691173 TI - Temporal evolution of traditional versus transformed ECG-based indexes in patients with induced myocardial ischemia. AB - The time course of changes in the electrocardiogram as a result of myocardial ischemia induced during prolonged coronary angioplasty has been studied. We have analyzed the electrocardiogram evolution during the occlusion in terms of the Ischemic Changes Sensor, which is a parameter that describes the capacity of different indexes to detect induced changes. Traditional indexes at specific time locations (ST level, T wave amplitude and position, and durations of QT interval and QRS complex) and global indexes (based on the Karhunen-Loeve transform as applied to the QRS complex, ST-T complex, ST segment and T wave) have been considered. The global indexes better detected ischemic changes than the traditional indexes. The most sensitive were the index for the ST-T complex (89%) in the Karhunen-Loeve transform-derived group and for the ST level (61%) in the traditional group. Changes in the ventricular repolarization period usually appeared earlier (77% of patients) than changes in the depolarization period (23% of patients). A similar percentage of patients exhibited the earliest ischemic changes in the T wave (41%) and in the ST segment (36%). The evolution of the Ischemic Changes Sensor parameters showed that the majority (60%) of the total changes occurred during the first minute of occlusion. The results suggest that the use of global electrocardiogram indexes better reflect ischemic changes than do traditional indexes, such as the ST segment deviation. PMID- 10691174 TI - Cardiac rupture and admission electrocardiography in acute anterior myocardial infarction: implication of ST elevation in aVL. AB - This study determines the usefulness of electrocardiography in the emergency room for assessing the risk of cardiac rupture after acute anterior myocardial infarction (MI). The presence of ST segment elevation on the admission 12-lead electrocardiography was evaluated in 325 consecutive anterior MI patients. A forward-stepwise logistic regression analysis for cardiac rupture was performed with the covariates of age, gender, hypertension, history of MI, reperfusion therapy by coronary angioplasty, and ST segment elevations in leads I, aVL, V1 V6. Cardiac rupture occurred in 16 patients, including 7 with left ventricular free wall rupture (FWR) and 9 with ventricular septal perforation (VSP). For FWR, ST elevation in lead aVL was the only independent predictor (odds ratio = 12.1, P = .0215). For VSP, female gender (odds ratio = 5.32, P = .0201) was the independent predictor. In conclusion, in patients with acute anterior MI, ST segment elevation in lead aVL on the admission electrocardiography is a significant risk factor for left ventricular FWR. PMID- 10691175 TI - Simulated torsade de pointes--the role of conduction defects and mechanism of QRS rotation. AB - A possible mechanism of torsade de pointes consisting of moving sites of reentry in the presence of disparate recovery of excitability has been previously proposed. This study evaluates the role of conduction defects in that mechanism. A computer model that simulated propagation, cycle length dependent recovery of excitability, and slow propagation during incomplete recovery and in conduction defects was used. Localized conduction defects consisting of slow propagation were shown to allow reentry at changing locations in the presence of uniform recovery properties. Later activation within defects resulted in later recovery, which permitted independent antegrade propagation adjacent to the defects. Retrograde propagation in the defects then resulted in reentry. The location of serial reentry changed because retrograde propagation and antegrade recovery had opposing directions and met distal to the origin of antegrade excitation. This mechanism was similar to that produced by disparate recovery and the combination of conduction defects and disparate recovery permitted the mechanism to occur with less marked disparity than otherwise required. The study also showed bidirectional serial reentry around a localized conduction defect or region of disparate recovery, which resulted in rotation of QRS peaks around the isoelectric line. The study provided evidence that either conduction defects or disparate recovery of excitability may be a substrate for torsade de pointes. It also indicated that combination of these factors might permit torsade de pointes when neither alone does so. This provides a possible explanation for the special propensity of quinidine and other drugs that slow conduction as well as prolong recovery to result in torsade de pointes. Findings also suggested a more explicit mechanism for rotation of QRS peaks about the electrocardiogram baseline than was previously available. PMID- 10691176 TI - Heart transplantation: influence of the surgical technique on sinus function. AB - This study analyzes the influence of the surgical technique on the development of sinus dysfunction. Cycle length, corrected sinus node recovery time, and sinoatrial conduction time were determined in 4 groups of dogs under the following conditions: group 1, controls; group 2, subjected to heterotopic heart transplantation with conservation of atrial anatomy; group 3, subjected to orthotopic heart transplantation by using a standard technique; and group 4, treated as in group 3, but with an ischemic time of 24 hours. The cycle length was prolonged in all the treated groups when compared with the controls (P < .01), an effect attributed to the disconnection of the autonomic nervous system during the transplantation procedure. The corrected sinus node recovery time and sinoatrial conduction time were significantly longer in all the animals in groups 3 and 4 when compared with those of groups 1 and 2 (P < .01); however, there were no significant differences between groups 3 and 4 or between groups 1 and 2. These results suggest that the atrial anatomy play a more relevant role than the ischemic time in the origin of sinus dysfunction occurring after orthotopic heart transplantation. PMID- 10691177 TI - Marked anterograde decremental conduction over a rapidly conducting accessory pathway after radiofrequency ablation. AB - We report on a patient with the Wolff-Parkinson-White syndrome who temporarily exhibited a marked anterograde decremental conduction over a rapidly conducting accessory atrioventricular pathway after successful radiofrequency ablation. By recording the intracardiac electrogram via the ablation catheter placed at the successful ablation site, we were able to exclude the possibility of the occurrence of anterograde decremental conduction in the atrial or ventricular myocardium between the accessory pathway and the recording electrodes. PMID- 10691178 TI - Simultaneous alterations of QRS configuration and tachycardia cycle length during radiofrequency ablation of idiopathic left ventricular tachycardia. AB - Idiopathic left ventricular tachycardia is characterized by a QRS morphology of right bundle branch block pattern and left axis deviation. Alterations in the QRS configuration and tachycardia cycle length, as well as shifting of the earliest activation site occurred after eliminating the original tachycardia by radiofrequency current in an 18-year-old man with idiopathic left ventricular tachycardia. Activation mapping and entrainment mapping during tachycardia identified 2 putative tachycardia exits, 15 mm apart. Elimination of both tachycardias was accomplished after applying radiofrequency current to each exit separately. We proposed that the first radiofrequency application might have altered the exit site and the zone of slow conduction adjacent to the exit site, such that the ventricular tachycardia had a different QRS morphology and became slower in this patient. PMID- 10691179 TI - Sinus escape-capture bigeminy and sinus extrasystolic bigeminy. AB - Blocking conduction between the sinus node and the atria (SA block) can be responsible for symptomatic rhythm problems. However, in atrial escape-capture bigeminy with SA block, when atrial escape P waves originate in a site within or close to the sinus node, the diagnosis of SA block is not easy. Electrocardiograms were selected from 7 people with atrial bigeminy because (1) all atrial deflections (P waves) were almost the same in shape and in length of PR intervals, (2) comparatively long PP intervals alternated with comparatively short PP intervals, and (3) occasionally the atrial bigeminy changed to normal regular sinus rhythm in which 2 or more sinus P waves were found in succession. An attempt is made to clarify the mechanism for these cases. When regular sinus rhythm changed to bigeminal rhythm, the long PP interval introduced the bigeminy in 3 cases, indicating the presence of "sinus" escape-capture bigeminy; whereas the short PP interval introduced the bigeminy in the other 4 cases, indicating the presence of "sinus" extrasystolic bigeminy. In cases of sinus escape-capture bigeminy associated with SA block, the cases may occasionally be diagnosed wrongly as ordinary sinus arrhythmia not associated with SA block. Therefore, it seems that sinus escape-capture bigeminy is not so rare as is generally believed. Patients with SA block often require implantation of the artificial pacemaker. Thus, the authors believe that differentiation of sinus escape-capture bigeminy from other forms of "sinus" bigeminy is clinically important. PMID- 10691180 TI - Concealed conduction in the reentrant pathway as a mechanism of stable ventricular quadrigeminy. AB - This is the first report on the stable occurrence of ventricular quadrigeminy as a manifestation of concealed bigeminy in a case of fixed and late coupled ventricular extrasystoles. A 46-year-old man is reported in whom the period of ventricular bigeminy alternated with the period of ventricular quadrigeminy. Coupling intervals of the extrasystoles were fixed and much longer than sinus QT intervals. When the heart rate is decreased, the period of bigeminy changed to the period of quadrigeminy without gradual decrease in coupling of the preceding extrasystoles. Once such a change occurred, stable quadrigeminy is maintained for a period. These findings suggest the possibility that concealed electrotonic conduction of blocked impulses and interference of conducted impulses may occur in the reentrant extrasystolic pathway as a mechanism of stable ventricular quadrigeminy. PMID- 10691181 TI - Affinity labeling of oxaloacetate decarboxylase by novel dichlorotriazine linked alpha-ketoacids. AB - The 4-aminophenyloxanilic acid and beta-mercaptopyruvic acid linked to the reactive diclorotriazine ring, were studied as active site-direct affinity labels towards oxaloacetate decarboxylase (EC 4.1.1.3, OXAD). Oxaloacetate decarboxylase when incubated with 4-aminophenyloxanilic-diclorotriazine (APOD) or beta mercaptopyruvic-diclorotriazine (MPD) at pH 7.0 and 25 degrees C shows a time dependent and concentration-dependent loss of enzyme activity. The inhibition was irreversible and activity cannot be recovered either by extensive dialysis or gel filtration chromatography. The enzyme inactivation following the Kitz & Wilson kinetics for time-dependent irreversible inhibition. The observed rate of enzyme inactivation (k(obs)) exhibits a non-linear dependence on APOD or MPD concentration with maximum rate of inactivation (k3) of 0.013 min(-1) and 0.0046 min(-1) and K(D) equal to 20.3 and 156 microM respectively. The inactivation of oxaloacetate decarboxylase by APOD and MPD is competitively inhibited by OXAD substrate and inhibitors, such as oxaloacetate, ADP and oxalic acid whereas Mn+2 enhances the rate of inactivation. The rate of inactivation of OXAD by APOD shows a pH dependence with an inflection point at 6.8, indicating a possible histidine derivatization by the label. These results show that APOD and MPD demonstrate the characteristics of an active-site probe towards the oxaloacetate binding site of oxaloacetate decarboxylase. PMID- 10691182 TI - Kinetics of inhibition of green crab (Scylla serrata) alkaline phosphatase by sodium (2,2'-bipyridine) oxodiperoxovanadate. AB - Green crab (Scylla serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, which catalyzes the nonspecific hydrolysis of phosphate monoesters. The kinetics of inhibition of the enzyme by sodium (2, 2'-bipyridine) oxodiperoxovanadate, pV(bipy), has been studied. The time course of the hydrolysis of p-nitrophenyl phosphate catalyzed by the enzyme in the presence of different pV(bipy) concentrations showed that at each pV(bipy) concentration, the rate decreased with increasing time until a straight line was approached, the straight line slopes are the same for all concentrations. The results suggest that the inhibition of the enzyme by pV(bipy) is a slow, reversible reaction with fractional remaining activity. The microscopic rate constants are determined for the reaction of inhibitor with the enzyme. PMID- 10691183 TI - Kinetics of formation of antibody-ferric porphyrin complex with peroxidase activity. AB - The antibody 2B4 combines with ferric mesoporphyrin to form an antibody-ferric mesoporphyrin complex which has a peroxidase activity. Formation of the complex was investigated by measuring the absorption in the Soret region after mixing the antibody and ferric mesoporphyrin. A rapid increase and a gradual decrease in the absorption were observed, and the respective first-order rate constants were obtained. From the dependence of values of the rate constants on the concentration of ferric mesoporphyrin, the complex formation was explained by a plausible mechanism, in which the antibody associated with ferric mesoporphyrin to form the first complex followed by a conformational change to the second complex. The first complex had almost the same peroxidase activity as that of the second complex. Our results suggests that the antibody acquires the peroxidase activity as soon as ferric mesoporphyrin is incorporated into its binding site, and that there will be no protein ligand to the iron center of ferric mesoporphyrin in the complex. PMID- 10691184 TI - Determination of amino acid sequences of two subunits in sarcosine oxidase from Corynebacterium sp. U-96. AB - The primary structures of the C and D subunits of sarcosine oxidase from Corynebacterium sp. U-96 were determined by sequencing the peptide fragments derived from their enzymatic digestions. The C and D subunits were shown to be composed of 199 and 92 residues, respectively. Each amino acid sequence showed a high homology with the sequence of the corresponding subunit from Corynebacterium sp. P-1. However, there were some differences between these two species, that is, four N-terminal residues were truncated in the C subunit, but six C-terminal residues were truncated in the D subunit. The D subunit contained three cysteine residues, but no disulfide bonds are in the subunit. Overall sequences of both subunit showed no homology with any other protein in the data base. PMID- 10691185 TI - Molecular composition of progenitor toxin produced by Clostridium botulinum type C strain 6813. AB - The molecular composition of the purified progenitor toxin produced by a Clostridium botulinum type C strain 6813 (C-6813) was analyzed. The strain produced two types of progenitor toxins (M and L). Purified L toxin is formed by conjugation of the M toxin (composed of a neurotoxin and a non-toxic nonhemagglutinin) with additional hemagglutinin (HA) components. The dual cleavage sites at loop region of the dichain structure neurotoxin were identified between Arg444-Ser445 and Lys449-Thr450 by the analyses of C-terminal of the light chain and N-terminal of the heavy chain. Analysis of partial amino acid sequences of fragments generated by limited proteolysis of the neurotoxin has shown to that the neurotoxin protein produced by C-6813 was a hybrid molecule composed of type C and D neurotoxins as previously reported. HA components consist of a mixture of several subcomponents with molecular weights of 70-, 55-, 33-, 26 through 21- and 17-kDa. The N-terminal amino acid sequences of 70-, 55-, and 26 through 21-kDa proteins indicated that the 70-kDa protein was intact HA-70 gene product, and other 55- and 26 through 21-kDa proteins were derived from the 70-kDa protein by modification with proteolysis after translation of HA-70 gene. Furthermore, several amino acid differences were exhibited in the amino acid sequence as compared with the deduced sequence from the nucleotide sequence of the HA-70 gene which was common among type C (strains C-St and C-468) and D progenitor toxins (strains D-CB16 and D-1873). PMID- 10691186 TI - Synthesis of aminobenzyltriethylenetetraaminohexaacetic acid: conjugation of the chelator to protein by an alkylamine linkage. AB - The conjugation of a chelating agent to an antibody as an anchoring site for a radionuclide is the first step in the successful preparation of a radiolabeled antibody for a diagnostic and therapeutic application. The high affinity of the protein bound chelator towards radionuclide ensures a higher selectivity in the delivery of the radionuclide to the targeted tissue. 4 Aminobenzylderivativetriethlenetetraaminohexaacetic acid (TTHA), a hexadentate chelating agent has been now prepared for conjugation with proteins in view of the higher affinity of TTHA metal ions as compared to DTPA. The latent crosslinking potential of alpha-hydroxy aldehydes has been used to conjugate the new chelating agent to proteins through an alkylamine linkage. On incubation of amino benzyl TTHA with glycoladehyde at neutral pH and room temperature, the reagent is converted to oxo ethyl amino benzyl TTHA. On addition of albumin to this reaction mixture, the oxo ethylamino benzyl TTHA generates reversible schiff base adducts with the amino groups of albumin. The reduction of the Schiff base adducts of the chelator with the protein by sodium cyanoborohydride stabilizes the schiff base adducts as stable alkylamine linkages. 4-Thiocyanatobenzyl TTHA has also been prepared and conjugated to albumin through a thiocarbamoyl linkage. Both preparations of TTHA conjugated albumin complexed with 99mTc and 111In, with high affinity and no decomposition of the complex was noticed for at least up to 6 hrs after the preparation. The radiolabels complexed with these TTHA -albumin conjugates could not be 'chased' out by free DTPA. A comparison of the biodistribution of 111In, bound to the TTHA conjugated through an alkylamine and a thiocarbamoyl linkage showed that 111In complexed with alkylamine linked TTHA was retained in blood to a level nearly 17% higher compared to that seen with thicarbamoyl linked TTHA, one hr after the injection into mice. Thus, the alkylamine linkage appears to be more stable under the in vivo conditions. The glycolaldehyde mediated alkylation procedure offers a mild, simple and rapid method for preparation of drug-protein (antibody) conjugates. PMID- 10691187 TI - Unfolding of hirudin characterized by the composition of denatured scrambled isomers. AB - The native core structure of hirudin, a thrombin specific inhibitor, contains 24 hydrogen bonds, two stretches of beta-sheet and three disulfide bonds. Hirudin unfolds in the presence of denaturant and thiol catalyst by shuffling its native disulfide bonds and converting to scrambled structures that consist of 11 identified isomers. The composition of scrambled isomers, which characterizes the structure of denatured hirudin, varies as a function of denaturing conditions. The unfolding pathway of hirudin has been constructed by quantitative analysis of scrambled isomers unfolded under increasing concentrations of various denaturants. The results demonstrate a progressive expansion of the polypeptide chain and the existence of a structurally defined stable intermediate along the pathway of unfolding. PMID- 10691188 TI - Identification of protein C epitopes altered during its nanoencapsulation. AB - Protein C is a plasmatic inhibitor which regulates the blood coagulation mechanism by modulating the anticoagulant response. The improvement of its bioavailability would be beneficial for the treatment of the disorders caused by its homozygous deficiency or by an other plasmatic inhibitor deficiency. In this context, the protein C encapsulation into biodegradable nanoparticles could be used to approach the problem. However, the method used to prepare the nanoparticles requires the use of ultrasonication and of an organic solvent such as methylene chloride which interferes with protein activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that neither ultrasonication nor methylene chloride, singly or in combination, led to protein C aggregation or cleavage. Thus, a binding study using an ELISA assay with four characterized monoclonal antibodies was carried out to identify the epitopes damaged by these experimental constraints. The correlation between the immunological assay and a functional one i.e. by the means of the assay of its anticoagulant activity (activated partial thromboplastin time) made it possible to show that protein C amino acids 166-169 of the activation peptide were probably altered after ultrasonication and methylene chloride treatment. Indeed, it is likely that these residues were no longer surface-exposed but had moved inside the protein core. PMID- 10691189 TI - Conformational changes at the active site of pantetheine hydrolase during denaturation by guanidine hydrochloride. AB - Conformational changes at the active site of pantetheine hydrolase (EC3.5.1.-) during guanidine hydrochloride (GndHCl) denaturation were investigated by UV and circular dichroism spectroscopy and by electron spin resonance spectroscopy, following the spectral behaviour of the nitroxide radicals (N-(1-oxyl-2,2,5,5, tetramethyl-3-pyrrolidinyl) iodacetamide) covalently linked to the two active site cysteine residues. At low denaturant concentrations (0.2 M) no conformational changes may be observed, whereas the catalytic activity, is strongly affected. The results indicate that the active site of pantetheine hydrolase is labile and unfolds under conditions in which no global tertiary structure modifications can be observed. PMID- 10691190 TI - Attractor control of the redox reactions of bovine cytochrome c. AB - Although the conformational changes accompanying the oxidation of ferrocytochrome c by the transfer of an electron to cytochrome a are small, they may contribute to the regulation of the electron transfer by transient storage of the liberated energy as strain and atomic vibrations. Both the electron transfer and the conformational changes seem to be controlled by an attractor, i.e. by a manifestation of a deterministic chaos. The putative attractor is regular and is, for the reaction involving the inner monomer of ferricytochrome c (I), of the order of 3.03 +/- 0.03. The conformational changes involving the outer monomer of ferricytochrome c (O) seem also to be controlled by a regular attractor, but its order is 4.2 +/- 0.2. The low order of the coupled reactions of electron transfer and conformational change suggests that it is essential to the electron transfer process in the respiratory chain. Since the order of attractors of other proteins correlates with the vectorial description of the function (1.0 for myoglobin, 2.0 for chymotrypsin and lysozyme, 3.0 for an abenzyme), the value for cyt. c indicates that not only the electron transfer, but also an additional reaction, e.g. the conformational change, are essential for the function of this protein. Hence, the study of protein attractors may yield information on important details, which could not be obtained by other methods. PMID- 10691191 TI - Differences of reconstitution process between tobacco mosaic virus and cucumber green mottle mosaic virus by synchrotron small angle X-ray scattering using low temperature quenching. AB - The differences of the reconstitution process of tobacco mosaic virus (TMV) and its mutant, cucumber green mottle mosaic virus (CGMMV) were investigated by the solution X-ray scattering measurements with the synchrotron radiation source using low-temperature quenching. The reconstitution in an aqueous solution is completely stopped below 5 degrees C. The TMV and CGMMV assembly was traced by the small-angle X-ray scattering (SAXS) measurements at 5 degrees C on a series of solutions prepared by low-temperature quenching after incubation at 20 degrees C for an appropriate interval between 0 and 60 min. The SAXS results were analyzed by the Guinier plot, the Kratky plot and the distance distribution function. The incubation of RNA and protein of CGMMV did not reconstitute at the initial reaction stages below 5 min and then began to reconstitute gradually. After 60 min, the radius of gyration for CGMMV reconstitution process reached almost the value for the initial stage of TMV reconstitution process. This is due to the fact the formation of double-layered disk in CGMMV protein is much slower than in TMV protein. PMID- 10691192 TI - Pivot residue: an analysis of domain motion in proteins. AB - In this study, we present an approach to identify some residues that represent the pivot points to experience conformational changes between open (unligand) and closed (ligand) forms of a protein. First, an angle, theta, formed by 4 consecutive Ca atoms in polypeptide backbones was introduced. The difference of this angle, deltatheta, from the equivalent residues between the open and the closed form was used to represent the local torsion changes in the protein structure, and the residue with the maximum among deltatheta was identified to be a pivot residue. We demonstrate the ability of our method by identifying the pivot residues from five proteins, Lysozyme mutates, Lactoferrin, Lay/Arg/Orn binding protein, Calmodulin and Catabolit gene activator protein. These pivot residues are located at the hinges in the proteins, they are hinge points for the domain motion. These examples also show that the pivot residues are useful to distinguish the mechanism between shear motion and hinge motion in a protein. PMID- 10691193 TI - New aspects in the molecular basis of polymer-associated infections due to staphylococci. AB - Coagulase-negative staphylococci, particularly Staphylococcus epidermidis, cause the majority of infections associated with both temporarily inserted and permanently implanted foreign bodies. In recent years, the pathogenesis of polymer-associated staphylococcal infection has become better understood, due in part to the characterization of further associated factors. The bacterial factors involved in the two phases of biofilm formation, i.e. the rapid adherence of bacteria to the polymer surface and the subsequent, more prolonged, accumulation phase, are presented in this review. The biofilm present on infected devices plays an important role in the pathogenicity of the infecting organism by protecting the embedded staphylococci and reducing the efficacy of host defenses and antimicrobial killing. PMID- 10691194 TI - New directions for future studies of community-acquired pneumonia: optimizing impact on patient care. PMID- 10691195 TI - Prospective study of community-acquired pneumonia of bacterial etiology in adults. AB - The aim of this study was to prospectively analyze the bacterial etiology of community-acquired pneumonia in adults in Spain. From May 1994 to February 1996, 392 episodes of CAP diagnosed in the emergency department of a 600-bed university hospital were studied. An etiological diagnosis based on noninvasive microbiological investigations was achieved in 228 cases (58%); 173 of these diagnoses were definitive and 55 probable. Streptococcus pneumoniae, which caused 23.9% of the episodes, was the predominant pathogen observed, followed by Chlamydia pneumoniae (13.5%) and Legionella pneumophila (12.5%). Other less frequent pathogens found were Haemophilus influenzae (2.3%), Pseudomonas aeruginosa (1.5%), Mycoplasma pneumoniae (1.3%), Coxiella burnetii (1%), Moraxella catarrhalis (2 cases), Nocardia spp. (2 cases), and Staphylococcus aureus (2 cases). Streptococcus pneumoniae was significantly more frequent in patients with underlying disease and/or age > or =60 years (28% vs. 13%, P = 0.002), while Legionella pneumophila was more frequent in patients below 60 years of age and without underlying disease (20% vs. 9%, P = 0.006). Likewise, Streptococcus pneumoniae and Legionella pneumophila were the most frequent etiologies in patients requiring admission to the intensive care unit, occurring in 29% and 26.3% of the patients, respectively. In addition to Streptococcus pneumoniae, other microorganisms such as Chlamydia pneumoniae and Legionella spp. should be seriously considered in adults with community-acquired pneumonia when initiating empiric treatment or ordering rapid diagnostic tests. PMID- 10691196 TI - Multiple mycoplasmal infections detected in blood of patients with chronic fatigue syndrome and/or fibromyalgia syndrome. AB - The aim of this study was to investigate the presence of different mycoplasmal species in blood samples from patients with chronic fatigue syndrome and/or fibromyalgia syndrome. Previously, more than 60% of patients with chronic fatigue syndrome/fibromyalgia syndrome were found to have mycoplasmal blood infections, such as Mycoplasma fermentans infection. In this study, patients with chronic fatigue syndrome/fibromyalgia syndrome were examined for multiple mycoplasmal infections in their blood. A total of 91 patients diagnosed with chronic fatigue syndrome/fibromyalgia syndrome and with a positive test for any mycoplasmal infection were investigated for the presence of Mycoplasma fermentans, Mycoplasma pneumoniae, Mycoplasma hominis and Mycoplasma penetrans in blood using forensic polymerase chain reaction. Among these mycoplasma-positive patients, infections were detected with Mycoplasma pneumoniae (54/91), Mycoplasma fermentans (44/91), Mycoplasma hominis (28/91) and Mycoplasma penetrans (18/91). Multiple mycoplasmal infections were found in 48 of 91 patients, with double infections being detected in 30.8% and triple infections in 22%, but only when one of the species was Mycoplasma pneumoniae or Mycoplasma fermentans. Patients infected with more than one mycoplasmal species generally had a longer history of illness, suggesting that they may have contracted additional mycoplasmal infections with time. PMID- 10691197 TI - Meropenem alone and in combination with vancomycin in experimental meningitis caused by a penicillin-resistant pneumococcal strain. AB - In a rabbit model of meningitis caused by a pneumococcus highly resistant to penicillin (MIC, 4 microg/ml), meropenem, a broad-spectrum carbapenem, was bactericidal (-0.48+/-0.14 deltalog10 cfu/ml h) and slightly superior to ceftriaxone (-0.34+/-0.23 deltalog10 cfu/ml x h) and vancomycin (-0.39+/-0.19 deltalog10 cfu/ml x h). Although the combination of vancomycin with ceftriaxone was significantly more active than ceftriaxone alone (-0.55+/-0.19 deltalog10 cfu/ml x h), only an insignificant gain was observed by the addition of vancomycin to meropenem (-0.55+/-0.28 deltalog10 cfu/ml x h). PMID- 10691198 TI - Mutations in the basal core promoter and precore/core gene of hepatitis B virus in patients with chronic active but not acute hepatitis B. AB - Around 5-10% of adults infected with hepatitis B virus (HBV) develop a chronic liver disease such as chronic active hepatitis (CAH), and it is unclear whether the clinical outcome depends solely on the immune response or whether viral factors also play a role. In this study, a search was therefore made for nucleotide mutations in the basic core promoter (BCP) and amino-acid substitutions in the precore/core region of HBV infecting patients with CAH or with acute hepatitis. The nucleotide sequences of the BCP and of the precore/core region were determined in virus from ten patients with CAH and ten with acute hepatitis. The precore/core sequences were also analysed in 14 additional patients (6 with CAH, 8 with acute hepatitis). In seven of the ten patients with CAH, five types of mutations were found in the BCP. Deletions in the precore/core region were observed in six patients. In all six patients where only the precore/core region was studied, amino-acid substitutions were present. In contrast, in the ten patients with acute hepatitis studied for BCP, a mutation was found in the BCP of one patient only. Of the 18 patients in whom the precore/core was studied, three had an amino-acid substitution in this region. The results show a clear link between CAH and both HBV BCP and precore/core region mutations, suggesting these mutations may play a role in the persistence of HBV infection. PMID- 10691199 TI - Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. AB - The aim of this study was to identify a sustainable cell line and culture method that could continuously provide a sufficient quantity of Toxoplasma gondii tachyzoites to serve the needs of a general hospital laboratory. Three continuous cell lines (HeLa, LLC and Vero) and three cell-culture methods (culture in conventional flasks, culture in membrane-based flasks and an automated culture system) were investigated. In multiplicity-of-infection and time-course experiments, HeLa was the cell line of choice. Harvests from HeLa cells had significantly higher tachyzoite yields than those from LLC cells (P<0.00005) or Vero cells (P<0.05). Membrane-based flasks gave higher yields (6.15x10(6) tachyzoites/ml) than conventional flasks (1-2x10(6) tachyzoites/ml) initially, but these were not sustained. The automated cell-culture system was unsuitable for parasite culture. Continuous passage in 25 cm2 flasks was successful, yielding 1x10(6) tachyzoites/ml; viability exceeded 90% after 96-120 h of infection throughout 38 passes, during which time the viability improved and the time to harvest became more consistent. Toxoplasma gondii grown in continuous culture in HeLa cells can provide a regular supply of viable tachyzoites. Demonstration that HeLa-derived tachyzoites could be used for the dye test confirms the potential of this in vitro system for use in general hospital laboratories. PMID- 10691200 TI - Efficacy of treatment with paromomycin, azithromycin, and nitazoxanide in a patient with disseminated cryptosporidiosis. AB - A 24-year-old HIV-positive heterosexual woman with disseminated cryptosporidiosis was monitored from January 1998 to May 1999. During this period, consecutive stool, sputum, and bile examinations showed the constant presence of Cryptosporidium oocysts. Although the patient was repeatedly treated with oral paromomycin and azithromycin and, finally, nitazoxanide, her condition continued to deteriorate. In order to monitor the in vitro susceptibility of the parasite, specimens from various sites were collected periodically. When the first clinical isolate was tested, the antimicrobial agents used (azithromycin at a concentration of 8 mg/l, paromomycin at of 1 mg/ml, and nitazoxanide at 10 mg/l) produced a decrease in parasite counts of 26.5%, 63.4%, and 67.2%, respectively. Subsequent isolates of Cryptosporidium parvum showed similar susceptibilities. This case demonstrates that failure of clinical treatment corresponded to inadequate growth inhibition of the parasite in vitro. PMID- 10691201 TI - Role of Haemophilus influenzae in intra-amniotic infection in patients with preterm rupture of membranes. AB - Haemophilus spp. were isolated from the amniotic fluid of eight of 110 consecutive women with preterm premature rupture of membranes (PROM) between 1992 and 1998. Isolates were nontypeable and classified according to biochemical test results as Haemophilus influenzae biotype I (n = 1), biotype II (n = 4), biotype III (n = 1) or biotype IV (n = 2). Primers recognizing specific sequences in the 16S rRNA of the cryptic genospecies of Haemophilus were employed to amplify the DNA of the eight isolates. One isolate classified as Haemophilus influenzae biotype II was confirmed as belonging to the genital cryptic species. Infectious morbidity occurred in five women and two newborns and was associated in most cases with biotype II. PMID- 10691202 TI - Pulmonary adiaspiromycosis in a patient with acquired immunodeficiency syndrome. AB - Adiaspiromycosis is a noninfectious, nonarthropod-transmitted fungal infection that occurs worldwide in lower vertebrates, especially rodents. However, humans may become accidental hosts. Reported here is a case of adiaspiromycosis of the lung in an HIV-positive, 40-year-old, bisexual man who first presented with cough and dyspnea. Cultures of a bronchoalveolar lavage and protected brush specimen revealed the presence of fungal elements that were identified as Emmonsia parva var. parva. The patient was successfully treated with amphotericin B and thereafter with fluconazole. This organism should be added to the list of pathogens that cause pulmonary infection in AIDS patients. PMID- 10691203 TI - Evaluation of a rapid immunochromatographic assay for the detection of Legionella antigen in urine samples. AB - A new immunochromatographic membrane assay for detecting Legionella pneumophila serogroup 1 antigen in urine samples (Binax Now Legionella Urinary Antigen Test; Binax, USA) was evaluated. Its sensitivity, specificity and level of agreement with the Binax enzyme immunoassay were compared using nonconcentrated and concentrated urine samples. The overall agreement between the two tests was 98.1%; the specificity of both was 100%. The sensitivity of the immunochromatographic assay was 55.5% in nonconcentrated urine and 97.2% in concentrated urine in comparison with the enzyme immunoassay, using concentrated urine as the reference test. This immunochromatographic assay screens successfully for Legionella pneumophila serogroup 1 soluble antigen in concentrated urine samples. PMID- 10691204 TI - Role of blood culture systems in the evaluation of epidemiological features of coagulase-negative staphylococcal bloodstream infection in critically ill patients. AB - The impact of blood culture systems on the detection of coagulase-negative staphylococcal bloodstream infections in critically ill patients prior to and following the introduction of the Bactec 9240 blood culture system (Becton Dickinson Diagnostic Instrument Systems, USA), which replaced the Bactec NR 730 (Becton Dickinson Diagnostic Instrument Systems), was investigated over a 3-year period. Following the introduction of the new culture system, the incidence of bloodstream infections doubled (P<0.001). Patient demographics, severity of illness, and mortality remained unchanged, while the annual standardized mortality ratio decreased significantly. These data suggest that blood culture systems may have a major impact on the perceived incidence of coagulase-negative staphylococcal bloodstream infections in this population. PMID- 10691205 TI - Evaluation of an automated system for identification of Enterobacteriaceae and nonfermenting bacilli. AB - The performance of the Vitek 2 (bioMerieux, France), a new fully automated system allowing rapid identification of microorganisms and susceptibility testing, and the Vitek 2 ID-GNB card (bioMerieux) was evaluated using 502 clinical isolates and stock collection strains of gram-negative rods belonging to 70 taxa. The number of isolates correctly identified to species and genus levels was 430 (85.7%) and 485 (96.6%), respectively. Clinical isolates of both Enterobacteriaceae and non-Enterobacteriaceae were better identified at the species level (95.3% and 74%, respectively) than stock collection strains (86.4% and 52.2%, respectively). The Vitek 2 ID-GNB card provides after 3 h a highly acceptable level of accuracy for identification of Enterobacteriaceae and non Enterobacteriaceae, including most atypical strains encountered in clinical situations. PMID- 10691206 TI - Comparative in vitro activity of quinolones against Stenotrophomonas maltophilia. AB - The susceptibility of 109 Stenotrophomonas maltophilia isolates, all characterized by pulsed-field gel electrophoresis, to nine quinolones was studied. Grepafloxacin, trovafloxacin, and moxifloxacin displayed similar intrinsic activities (MIC90, 0.5 microg/ml), which were lower than those of ofloxacin and ciprofloxacin (MIC90, 4 microg/ml), norfloxacin (MIC90, 64 microg/ml), and nalidixic acid (MIC90, 32 microg/ml). Nalidixic acid was generally one- to twofold dilutions more active than norfloxacin. According to the criteria of the National Committee for Clinical Laboratory Standards (NCCLS), the percentage of isolates susceptible to ciprofloxacin (breakpoint < or = 1 microg/ml) was 76.1%. Using the NCCLS breakpoint for comparative purposes, the percentage of isolates susceptible to grepafloxacin, moxifloxacin, and trovafloxacin was 95.4, 96.4, and 96.4%, respectively. These results indicate that new quinolones may potentially be used for the management of Stenotrophomonas maltophilia infections. PMID- 10691207 TI - Comparison of two automated systems for the isolation of mycobacteria from clinical specimens. AB - The Bactec MGIT 960 (Becton-Dickinson, UK) automated mycobacterial liquid culture system was compared with the Bactec 9000 MB (Becton-Dickinson) in order to assess ease of use, diagnostic reliability and safety features. One thousand twenty-nine clinical specimens were cultured in parallel, yielding a total of 125 (12.1%) mycobacterial isolates, including 71 Mycobacterium tuberculosis and 18 Mycobacterium avium. The Bactec MGIT 960 demonstrated a mycobacterial recovery rate and speed of detection equivalent to that of the Bactec 9000 MB for clinically important isolates. The Bactec MGIT 960 integrates smoothly into laboratory workflow, does not require needle inoculation and has a much larger capacity than the Bactec 9000 MB. PMID- 10691208 TI - Antimicrobial susceptibilities and biotypes of Arcanobacterium haemolyticum blood isolates. AB - Isolates obtained from the blood of ten patients with Arcanobacterium haemolyticum septicaemia were biotyped as smooth or rough using morphological and biochemical criteria, and their susceptibilities to 18 antibacterial agents were determined. Nine of the clinical cases included here have not been reported previously and are discussed in brief. One of the strains was highly resistant to macrolides and clindamycin. With one exception, the strains belonged to the smooth biotype. The data presented here indicates that the treatment of systemic Arcanobacterium haemolyticum infections should be based on the antibacterial susceptibility profiles of individual strains and on the site of the infection. PMID- 10691209 TI - Bacteriology and clinical course of camel-bite wound infections. PMID- 10691210 TI - Lack of effect of antibiotic resistance on susceptibility of microorganisms to chlorhexidine gluconate or povidone iodine. PMID- 10691211 TI - Rostral spread of epidural morphine: the expected and the unexpected. PMID- 10691212 TI - The direct search procedure: a new approach to evaluating clinical regimens. PMID- 10691213 TI - Dopamine: one size does not fit all. PMID- 10691214 TI - Improving splanchnic perfusion during cardiopulmonary bypass. PMID- 10691215 TI - Preemptive hyperalgesia, not analgesia? PMID- 10691216 TI - Lumbar epidural morphine in humans and supraspinal analgesia to experimental heat pain. AB - BACKGROUND: Epidural administration of morphine is a common analgesic technique to manage pain. Morphine spreads from the epidural space to the cerebrospinal fluid and then rostrally, causing side effects mediated by the brain stem. However, data on the rostral spread of morphine-mediated analgesia are sparse. This study examined the rostral spread of analgesic effects on heat and electrical pain after epidural administration of morphine. METHODS: In a randomized, double-blinded, placebo-controlled, crossover study, 5 mg morphine or saline placebo were injected into the lumbar epidural space in nine healthy volunteers. Correct needle placement was confirmed with fluoroscopy. Analgesia to experimental nociceptive heat and electrical stimuli was measured at lumbar (L4), thoracic (T10), cervical (C2), and trigeminal (V2) levels before and 2, 5, 10, and 24 h after epidural injection. Plasma samples for assaying morphine concentrations were drawn before and after each analgesic evaluation. RESULTS: Epidural morphine significantly attenuated experimental heat pain at all dermatomes tested compared with saline placebo. Analgesic effects were significant at L4 after 2, 5, and 10 h, at T10 after 5, 10, and 24 h, and at V2 after 10 h. Electrical pain was attenuated at the lumbar and thoracic but not at the cervical dermatome. Analgesic effects were significant at L4 after 2, 5, and 10 h and at T10 after 5 and 10 h. Morphine plasma concentrations were below the detection limit (1 ng/ml) in eight of the nine subjects 10 h after epidural injection. CONCLUSIONS: Lumbar epidural injection of morphine attenuated cutaneous heat pain up to the trigeminal dermatome during a 24-h observation period. In a clinical context, this implies that some types of pain may be attenuated up to the supraspinal level after lumbar epidural administration of morphine. PMID- 10691217 TI - A direct search procedure to optimize combinations of epidural bupivacaine, fentanyl, and clonidine for postoperative analgesia. AB - BACKGROUND: The authors applied an optimization model (direct search) to find the optimal combination of bupivacaine dose, fentanyl dose, clonidine dose, and infusion rate for continuous postoperative epidural analgesia. METHODS: One hundred ninety patients undergoing 48-h thoracic epidural analgesia after major abdominal surgery were studied. Combinations of the variables of bupivacaine dose, fentanyl dose, clonidine dose, and infusion rate were investigated to optimize the analgesic effect (monitored by verbal descriptor pain score) under restrictions dictated by the incidence and severity of side effects. Six combinations were empirically chosen and investigated. Then a stepwise optimization model was applied to determine subsequent combinations until no decrease in the pain score after three consecutive steps was obtained. RESULTS: Twenty combinations were analyzed. The optimization procedure led to a reduction in the incidence of side effects and in the mean pain scores. The three best combinations of bupivacaine dose (mg/h), fentanyl dose (microg/h), clonidine dose (microg/h), and infusion rate (ml/h) were: 9-21-5-7, 8-30-0-9, and 13-25-0-9, respectively. CONCLUSIONS: Given the variables investigated, the aforementioned combinations may be the optimal ones to provide postoperative analgesia after major abdominal surgery. Using the direct search method, the enormous number of possible combinations of a therapeutic strategy can be reduced to a small number of potentially useful ones. This is accomplished using a scientific rather than an arbitrary procedure. PMID- 10691218 TI - Pharmacokinetics of dopamine in healthy male subjects. AB - BACKGROUND: Dopamine is an agonist of alpha, beta, and dopaminergic receptors with varying hemodynamic effects depending on the dose of drug being administered. The purpose of this study was to measure plasma concentrations of dopamine in a homogeneous group of healthy male subjects to develop a pharmacokinetic model for the drug. Our hypothesis was that dopamine concentrations can be predicted from the infusion dose using a population-based pharmacokinetic model. METHODS: Nine healthy male volunteers aged 23 to 45 yr were studied in a clinical research facility within our academic medical center. After placement of venous and arterial catheters, dopamine was infused at 10 microg x kg(-1) x min(-1) for 10 min, followed by a 30-min washout period. Subsequently, dopamine was infused at 3 microg x kg(-1) x min(-1) for 90 min, followed by another 30-min washout period. Timed arterial blood samples were centrifuged, and the plasma was analyzed by high-performance liquid chromatography. Mixed-effects pharmacokinetic models using NONMEM software (NONMEM Project Group, University of California, San Francisco, CA) were used to determine the optimal compartmental pharmacokinetic model for dopamine. RESULTS: Plasma concentrations of dopamine varied from 12,300 to 201,500 ng/l after 10 min of dopamine infusion at 10 microg x kg(-1) x min(-1). Similarly, steady-state dopamine concentrations varied from 1,880 to 18,300 ng/l in these same subjects receiving 3-microg x kg(-1) x min(-1) infusions for 90 min. A two-compartment model adjusted for body weight was the best model based on the Schwartz-Bayesian criterion. CONCLUSIONS: Despite a homogeneous population of healthy male subjects and weight-based dosing, there was 10- to 75-fold intersubject variability in plasma dopamine concentrations, making standard pharmacokinetic modeling of less utility than for other drugs. The data suggest marked intraindividual and interindividual variability in dopamine distribution and/or metabolism. Thus, plasma dopamine concentrations in patients receiving dopamine infusion at identical rates may vary profoundly. Our data suggest that dosing dopamine based on body weight does not yield predictable blood concentrations. PMID- 10691219 TI - Efficacy of neurolytic celiac plexus block in varying locations of pancreatic cancer: influence on pain relief. AB - BACKGROUND: Neurolytic celiac plexus block (NCPB) is an effective way of treating severe pain in some patients with pancreatic malignancy. However, there are no studies to date that evaluate the effectiveness of NCPB related to the site of primary pancreas cancer. The aim of the study was to assess the effectiveness of NCPB in pancreatic cancer pain, depending on the location of the pancreatic tumor. METHODS: The prospective study was conducted in 50 consecutive patients diagnosed with pancreatic cancer. The patients were categorized into two different groups depending on tumor localization: group 1: patients with the cancer of the head of the pancreas and group 2: patients with the cancer of the body and tail of the pancreas. The qualitative and quantitative pain analyses were performed before and after NCPB. The patients underwent prognostic celiac plexus block with bupivacaine, followed by neurolysis during fluoroscopic control within the next 24 h. RESULTS: After NCPB, 37 patients (74%) had effective pain relief during the first 3 months or until death. Of the 37 patients who had effective pain relief, 33 (92%) were from group 1 and 4 (29%) were from group 2. In the remaining 13 patients (3 patients from group 1 and 10 patients from group 2), pain relief after NCPB was not satisfactory. Those patients were scheduled for repeated retrocrural neurolysis during computed tomography control. Computed tomography showed massive growth of the tumor around the celiac axis with metastases. After repeated neurolysis, pain relief clinically still was not satisfactory, necessitating additional opioid treatment. CONCLUSION: In this study, unilateral transcrural celiac plexus neurolysis has been shown to provide effective pain relief in 74% of patients with pancreatic cancer pain. Neurolysis was more effective in cases with tumor involving the head of the pancreas. In the cases with advanced tumor proliferation, regardless of the technique used, the analgesic effects of NCPB were not satisfactory. PMID- 10691220 TI - Preemptive intravenous morphine-6-glucuronide is ineffective for postoperative pain relief. AB - BACKGROUND: Morphine-6-glucuronide (M-6-G), a major metabolite of morphine, is reported to be more potent than morphine when administered intrathecally; however, its efficiency remains under debate when administered intravenously. This study was designed to assess the analgesic efficiency of intravenous M-6-G for the treatment of acute postoperative pain. METHODS: After informed consent was obtained, 37 adults (American Society of Anesthesiologists physical status I II) who were scheduled for elective open knee surgery were enrolled in the study. General anesthesia was induced with thiopental, alfentanil, and vecuronium and was maintained with a mixture of nitrous oxide/isoflurane and bolus doses of alfentanil. At skin closure, patients were randomized into three groups: (1) morphine group (n = 13), which received morphine 0.15 mg/kg; (2) M-6-G group (n = 12), which received M-6-G 0.1 mg/kg; and (3) placebo group (n = 12), which received saline. At the time of extubation, plasma concentration of morphine and M-6-G was measured. Postoperative analgesic efficiency was assessed by the cumulative dose of morphine delivered by patient-controlled analgesia. Opioid related side effects were also evaluated. RESULTS: No difference was noted in patient characteristics and opioid-related side effects. Morphine requirements (mean +/- SD) during the first 24 h in the M-6-G group (41+/-9 mg) and the placebo group (49+/-8 mg) were significantly greater (P<0.05) compared with the morphine group (29+/-8 mg). CONCLUSION: A single intravenous bolus dose of M-6-G was found to be ineffective in the treatment of acute postoperative pain. This might be related to the low permeability of the blood-brain barrier for M-6-G. PMID- 10691221 TI - Low-dose clonidine and neostigmine prolong the duration of intrathecal bupivacaine-fentanyl for labor analgesia. AB - BACKGROUND: Intrathecal (IT) opioid and local anesthetic combinations are popular for labor analgesia because of rapid, effective pain relief, but the duration of analgesia is limited. This study was undertaken to determine whether the addition of clonidine and neostigmine to IT bupivacaine-fentanyl would increase the duration of analgesia without increasing side effects for patients in labor. METHODS: Forty-five healthy parturients in active labor were randomized to receive a 2-ml IT dose of one of the following dextrose-containing solutions using the combined spinal-epidural technique: (1) bupivacaine 2.5 mg and fentanyl 25 microg (BF); (2) BF plus clonidine 30 microg (BFC); or (3) BFC plus neostigmine 10 microg (BFCN). Pain, sensory levels, motor block, side effects, maternal vital signs, and fetal heart rate were systematically assessed. RESULTS: Patients administered BFCN had significantly longer analgesia (165+/-32 min) than those who received BF (90+/-21 min; P<0.001) or BFC (123+/-21 min; P<0.001). Pain scores, block characteristics, maternal vital signs, Apgar scores, maternal satisfaction, and side effects were similar among groups except for nausea, which was significantly greater in the BFCN group (P<0.05 as compared with BFC). CONCLUSIONS: The addition of clonidine and neostigmine significantly increased the duration of analgesia from IT bupivacaine-fentanyl during labor, but neostigmine caused more nausea. Although serious side effects were not observed in this study, safety must be further addressed before the routine use of multiple IT drugs is advocated. PMID- 10691222 TI - Bedside assessment of cerebral blood flow by double-indicator dilution technique. AB - BACKGROUND: Currently, quantitative measurement of global cerebral blood flow (CBF) at bedside is not widely performed. The aim of the present study was to evaluate a newly developed method for bedside measurement of CBF based on thermodilution in a clinical setting. METHODS: The investigation was performed in 14 anesthetized patients before coronary bypass surgery. CBF was altered by hypocapnia, normocapnia, and hypercapnia. CBF was measured simultaneously by the Kety-Schmidt inert-gas technique with argon and a newly developed transcerebral double-indicator dilution technique (TCID). For TCID, bolus injections of ice cold indocyanine green were performed via a central venous line, and the resulting thermo-dye dilution curves were recorded simultaneously in the aorta and the jugular bulb using combined fiberoptic thermistor catheters. CBF was calculated from the mean transit times of the indicators through the brain. RESULTS: Both methods of measurement of CBF indicate a decrease during hypocapnia and an increase during hypercapnia, whereas cerebral metabolic rate remained unchanged. Bias between CBF(TCID) and CBFargon was -7.1+/-2.2 (SEM) ml x min(-1) x 100 g(-1); precision (+/- 2 x SD of differences) between methods was 26.6 ml x min(-1) x 100 g(-1). CONCLUSIONS: In the clinical setting, TCID was feasible and less time-consuming than alternative methods. The authors conclude that TCID is an alternative method to measure global CBF at bedside and offers a new opportunity to monitor cerebral perfusion of patients. PMID- 10691223 TI - Pharmacokinetics of rapacuronium in infants and children with intravenous and intramuscular administration. AB - BACKGROUND: A nondepolarizing muscle relaxant with an onset and offset profile similar to succinylcholine is desirable for pediatric anesthesia. The onset and offset of rapacuronium are rapid in children. In the current study, the authors determined its pharmacokinetic characteristics in children. In addition to administering rapacuronium by the usual intravenous route, the authors also gave rapacuronium intramuscularly to determine uptake characteristics and bioavailability. METHODS: Forty unpremedicated patients aged 2 months to 3 yr were anesthetized with halothane, 0.82-1.0% end-tidal concentration. When anesthetic conditions were stable, rapacuronium was injected either into a peripheral vein (2 mg/kg for infants, 3 mg/kg for children) or a deltoid muscle (2.8 mg/kg for infants, 4.8 mg/kg for children). Four venous plasma samples were obtained from each subject 2-240 min after rapacuronium administration. A mixed effects population pharmacokinetic analysis was applied to these values to determine bioavailability, absorption rate constant, and time to peak plasma concentration with intramuscular administration. RESULTS: Plasma clearance was 4.77 ml x kg(-1) x min(-1) + 8.48 ml/min. Intramuscular bioavailability averaged 56%. Absorption from the intramuscular depot had two rate constants: 0.0491 min( 1) (72.4% of absorbed drug) and 0.0110 min(-1) (27.6% of the absorbed drug). Simulation indicated that plasma concentration peaks 4.0 and 5.0 min after intramuscular rapacuronium in infants and children, respectively, and that, at 30 min, less than 25% of the administered dose remains to be absorbed from the intramuscular depot. CONCLUSIONS: In infants and children, rapacuronium's clearance and steady state distribution volume are less than in adults. After intramuscular administration, bioavailability is 56%, and plasma rapacuronium concentrations peak within 4 or 5 min. PMID- 10691224 TI - Walking with labor epidural analgesia: the impact of bupivacaine concentration and a lidocaine-epinephrine test dose. AB - BACKGROUND: Regional analgesia techniques for labor that permit ambulation are popular among parturients. This study evaluated the influence of bupivacaine bolus concentration and a 3-ml 1.5% lidocaine-epinephrine test dose, on analgesic effectiveness and the ability to walk after block placement. METHODS: Using a randomized double-blind study design, epidural analgesia was initiated in 60 parturients undergoing labor as follows: Group TD/B.0625 received a 3-ml lidocaine-epinephrine test dose + 12 ml bupivacaine, 0.0625%; group TD/B.125 received a 3-ml test dose + 12 ml bupivacaine, 0.125%; group B.0625 received 15 ml bupivacaine, 0.0625% (no test dose); and group B.125 received 15 ml bupivacaine, 0.125% (no test dose). Initial boluses in all groups contained 10 microg sufentanil. Bupivacaine, 0.0625%, with 0.33 microg/ml sufentanil was infused throughout labor at 13.5-15 ml/h. Analgesia balance, proprioception, motor block, and patient ability to stand and walk were evaluated at various intervals. RESULTS: A bolus of 0.125% bupivacaine containing sufentanil, without a previous test dose, proved to be optimal with respect to analgesia and early ambulation. When a test dose was given before bupivacaine, 0.125%, fewer women walked within 1 h of block placement. Bupivacaine, 0.0625%, with sufentanil, with or without a test dose, provided inadequate analgesia, necessitating additional bupivacaine, which impaired the ability to walk. A high percentage of women in all groups (73-93%) walked at some stage during labor. CONCLUSIONS: Omitting a lidocaine-epinephrine test dose and using 0.125% bupivacaine for the initial bolus should permit ambulation in the early postblock period for most parturients who elect this option. PMID- 10691225 TI - The pharmacodynamic effect of a remifentanil bolus on ventilatory control. AB - BACKGROUND: In doses typically administered during conscious sedation, remifentanil may be associated with ventilatory depression. However, the time course of ventilatory depression after an initial dose of remifentanil has not been determined previously. METHODS: In eight healthy volunteers, the authors determined the time course of the ventilatory response to carbon dioxide using the dual isohypercapnic technique. Subjects breathed via mask from a to-and-fro circuit with variable carbon dioxide absorption, allowing the authors to maintain end-tidal pressure of carbon dioxide (PET(CO2)) at approximately 46 or 56 mm Hg (alternate subjects). After 6 min of equilibration, subjects received 0.5 microg/kg remifentanil over 5 s, and minute ventilation (V(E)) was recorded during the next 20 min. Two hours later, the study was repeated using the other carbon dioxide tension (56 or 46 mm Hg). The V(E) data were used to construct two point carbon dioxide response curves at 30-s intervals after remifentanil administration. Using published pharmacokinetic values for remifentanil and the method of collapsing hysteresis loops, the authors estimated the effect-site equilibration rate constant (k(eo)), the effect-site concentration producing 50% respiratory depression (EC50), and the shape parameter of the concentration response curve (gamma). RESULTS: The slope of the carbon dioxide response decreased from 0.99 [95% confidence limits 0.72 to 1.26] to a nadir of 0.27 l x min(-1) x mm Hg(-1) [-0.12 to 0.66] 2 min after remifentanil (P<0.001); within 5 min, it recovered to approximately 0.6 l x min(-1) x mm Hg(-1), and within 15 min of injection, slope returned to baseline. The computed ventilation at PET = 50 mm Hg (VE50) decreased from 12.9 [9.8 to 15.9] to 6.1 l/min [4.8 to 7.4] 2.5 min after remifentanil injection (P<0.001). This was caused primarily by a decrease in tidal volume rather than in respiratory rate. Estimated pharmacodynamic parameters based on computed mean values of VE50 included k(eo) = 0.24 min(-1) (T1/2 = 2.9 min), EC50 = 1.12 ng/ml, and gamma = 1.74. CONCLUSIONS: After administration of 0.5 microg/kg remifentanil, there was a decrease in slope and downward shift of the carbon dioxide ventilatory response curve. This reached its nadir approximately 2.5 min after injection, consistent with the computed onset half-time of 2.9 min. The onset of respiratory depression appears to be somewhat slower than previously reported for the onset of remifentanil-induced electroencephalographic slowing. Recovery of ventilatory drive after a small dose essentially was complete within 15 min. PMID- 10691226 TI - Comparison of plasma compartment versus two methods for effect compartment- controlled target-controlled infusion for propofol. AB - BACKGROUND: Target-controlled infusion (TCI) systems can control the concentration in the plasma or at the site of drug effect. A TCI system that targets the effect site should be able to accurately predict the time course of drug effect. The authors tested this by comparing the performance of three control algorithms: plasmacontrol TCI versus two algorithms for effect-site control TCI. METHODS: One-hundred twenty healthy women patients received propofol via TCI for 12-min at a target concentration of 5.4 microg/ml. In all three groups, the plasma concentrations were computed using pharmacokinetics previously reported. In group I, the TCI device controlled the plasma concentration. In groups II and III, the TCI device controlled the effect-site concentration. In group II, the effect site was computed using a half-life for plasma effect-site equilibration (t1/2k(eo)) of 3.5 min. In group III, plasma effect-site equilibration rate constant (k(eo)) was computed to yield a time to peak effect of 1.6 min after bolus injection, yielding a t1/2keo of 34 s. the time course of propofol was measured using the bispectral index. Blood pressure, ventilation, and time of loss of consciousness were measured. RESULTS: The time course of propofol drug effect, as measured by the bispectral index, was best predicted in group III. Targeting the effect-site concentration shortened the time to loss of consciousness compared with the targeting plasma concentration without causing hypotension. The incidence of apnea was less in group III than in group II. CONCLUSION: Effect compartment-controlled TCI can be safely applied in clinical practice. A biophase model combining the Marsh kinetics and a time to peak effect of 1.6 min accurately predicted the time course of propofol drug effect. PMID- 10691227 TI - Critical oxygen delivery in conscious humans is less than 7.3 ml O2 x kg(-1) x min(-1). AB - BACKGROUND: The "critical" level of oxygen delivery (DO2) is the value below which DO2 fails to satisfy the metabolic need for oxygen. No prospective data in healthy, conscious humans define this value. The authors reduced DO2 in healthy volunteers in an attempt to determine the critical DO2. METHODS: With Institutional Review Board approval and informed consent, the authors studied eight healthy, conscious volunteers, aged 19-25 yr. Hemodynamic measurements were obtained at steady state before and after profound acute isovolemic hemodilution with 5% albumin and autologous plasma, and again at the reduced hemoglobin concentration after additional reduction of DO2 by an infusion of a beta adrenergic antagonist, esmolol. RESULTS: Reduction of hemoglobin from 12.5+/-0.8 g/dl to 4.8+/-0.2 g/dl (mean +/- SD) increased heart rate, stroke volume index, and cardiac index, and reduced DO2 (14.0+/-2.9 to 9.9+/-20 ml O2 x kg(-1) x min( 1); all P<0.001). Oxygen consumption (VO2; 3.0+/-0.5 to 3.4+/-0.6 ml O2 x kg(-1) x min(-1); P<0.05) and plasma lactate concentration (0.50+/-0.10 to 0.62+/-0.16 mM; P<0.05; n = 7) increased slightly. Esmolol decreased heart rate, stroke volume index, and cardiac index, and further decreased DO2 (to 7.3+/-1.4 ml O2 x kg(-1) x min(-1); all P<0.01 vs. before esmolol). VO2 (3.2+/-0.6 ml O2 x kg(-1) x min(-1); P>0.05) and plasma lactate (0.66+/-0.14 mM; P>0.05) did not change further. No value of plasma lactate exceeded the normal range. CONCLUSIONS: A decrease in DO2 to 7.3+/-1.4 ml O2 x kg(-1) min(-1) in resting, healthy, conscious humans does not produce evidence of inadequate systemic oxygenation. The critical DO2 in healthy, resting, conscious humans appears to be less than this value. PMID- 10691228 TI - Female gender associates with increased duration of intubation and length of stay after coronary artery surgery. CABG Clinical Benchmarking Database Participants. AB - BACKGROUND: Females have worse outcome than do males after coronary artery bypass grafting; however, gender effects on length of stay (LOS) outcomes, such as duration of intubation or intensive care unit (ICU) LOS, have not been evaluated previously. The authors hypothesized that adjustment for pertinent preoperative covariates would eliminate any significant effect of gender on duration of intubation, LOS in the ICU after extubation, total ICU LOS, postoperative (exclusive of ICU) LOS, or total postoperative LOS. METHODS: Patients undergoing elective or urgent primary coronary artery bypass grafting surgery at 51 academic health centers in 1995 and 1997 were studied. Unique multivariable statistical models were developed for duration of intubation, ICU LOS after extubation, total ICU LOS, and postoperative (exclusive of ICU and total) LOS to test for independent associations with gender. Preoperative but not intraoperative or postoperative variables were included in the model. P> or =0.01 was considered significant. RESULTS: All LOSs were of significantly longer duration in females than in males in both the 1995 (n = 1,064) and 1997 (n = 910) data collections. After covariate adjustment, female sex remained associated with significantly longer duration ICU LOS and total postoperative LOS in both the 1995 (female:male ratios 1.30:1 and 1.13:1, respectively) and the 1997 (female:male ratios 1.19:1 and 1.12:1, respectively) data sets. After covariate adjustment, duration of intubation and ICU LOS after extubation were of significantly longer duration in women than men in 1995 (female:male ratios 1.22:1 and 1.39:1, respectively), but the differences were not significant in 1997. CONCLUSIONS: Even in the context of accelerated recovery programs, these analyses show that female sex has powerful associations with increased LOS intervals for coronary artery bypass grafting surgery, even after adjustment for preoperative covariates. These effects could result from differences in the ways in which men and women respond to coronary artery disease, anesthesia, and coronary artery bypass grafting surgery, or to bias on the part of healthcare workers. PMID- 10691229 TI - Risk of surgery and anesthesia for ischemic stroke. AB - BACKGROUND: The goal of this study was to determine if the combination of surgery and anesthesia is an independent risk factor for the development of incident (first-time) ischemic stroke. METHODS: All residents of Rochester, MN, with incident ischemic stroke from 1960 through 1984 (1,455 cases and 1,455 age- and gender-matched controls) were used to identify risk factors associated with ischemic stroke. Cases and controls undergoing surgery involving general anesthesia or central neuroaxis blockade before their stroke/index date of diagnosis were identified. A conditional logistic regression model was used to estimate the odds ratio of surgery and anesthesia for ischemic stroke while adjusting for other known risk factors. RESULTS: There were 59 cases and 17 controls having surgery within 30 days before their stroke/index date. After adjusting for previously identified risk factors, surgery within 30 days before the stroke/index date (perioperative period) was found to be an independent risk factor for stroke (P<0.001; odds ratio, 3.9; 95% confidence interval, 2.1-7.4). In an analysis that excluded matched pairs where the case and/or control underwent surgery considered "high risk" for stroke (cardiac, neurologic, or vascular procedures), "non-high-risk surgery" was also found to be an independent risk factor for perioperative stroke (P = 0.002; odds ratio, 2.9; 95% confidence interval, 1.5-5.7). CONCLUSION: Our results suggest that there is an increased risk of ischemic stroke in the 30 days after surgery and anesthesia. This risk remains elevated even after excluding surgeries (cardiac, neurologic, and vascular surgeries) considered to be high risk for ischemic stroke. PMID- 10691230 TI - Comparison of intravenous or epidural patient-controlled analgesia in the elderly after major abdominal surgery. AB - BACKGROUND: Patient-controlled analgesia (PCA) with intravenous morphine and patient-controlled epidural analgesia (PCEA), using an opioid either alone or in combination with a local anesthetic, are two major advances in the management of pain after major surgery. However, these techniques have been evaluated poorly in elderly people. This prospective, randomized study compared the effectiveness on postoperative pain and safety of PCEA and PCA after major abdominal surgery in the elderly patient. METHODS: Seventy patients older than 70 yr of age and undergoing major abdominal surgery were assigned randomly to receive either combined epidural analgesia and general anesthesia followed by postoperative PCEA, using a mixture of 0.125% bupivacaine and sufentanil (PCEA group), or general anesthesia followed by PCA with intravenous morphine (PCA group). Pain intensity was tested three times daily using a visual analog scale. Postoperative evaluation included mental status, cardiorespiratory and gastrointestinal functions, and patient satisfaction scores. RESULTS: Pain relief was better at rest (P = 0.001) and after coughing (P = 0.002) in the PCEA group during the 5 postoperative days. Satisfaction scores were better in the PCEA group. Although incidence of delirium was comparable in the PCA and PCEA groups (24% vs. 26%, respectively), mental status was improved on the fourth and fifth postoperative days in the PCEA group. The PCEA group recovered bowel function more quickly than did the PCA group. Cardiopulmonary complications were similar in the two groups. CONCLUSION: After major abdominal surgery in the elderly patient, patient controlled analgesia, regardless of the route (epidural or parenteral), is effective. The epidural route using local anesthetics and an opioid provides better pain relief and improves mental status and bowel activity. PMID- 10691231 TI - Hypoxic brain tissue following subarachnoid hemorrhage. AB - BACKGROUND: Subarachnoid hemorrhage can lead to cerebral ischemia and irreversible brain injury. The purpose of this study was to determine whether subarachnoid hemorrhage produces changes in brain tissue oxygen pressure, carbon dioxide pressure, or pH during surgery for cerebral aneurysm clipping. METHODS: After institutional review board approval and patient consent, 30 patients undergoing craniotomy for cerebral aneurysm clipping were studied, 15 without and 15 with subarachnoid hemorrhage. Patients with subarachnoid hemorrhage were prospectively separated into groups with modest (Fisher grade 1 or 2; n = 8) and severe bleeds (Fisher grade 3; n = 7). After a craniotomy, a probe was inserted into cortex tissue supplied by the artery associated with the aneurysm. Baseline measures were made in the presence of a 4% end-tidal desflurane level. The end tidal desflurane level was increased to 9% before clipping of the aneurysm, and a second tissue measurement was made. RESULTS: The median time of surgery after subarachnoid hemorrhage was 2 days, ranging from 1 to 13 days. During baseline anesthesia, brain tissue oxygen pressure was 17+/-9 mm Hg (mean +/- SD) in control patients, 13+/-9 mm Hg in those with Fisher grade 1 or 2 hemorrhage, and 7+/-6 mm Hg in those with Fisher grade 3 hemorrhage (P<0.05 compared with control). Brain tissue pH was 7.10+/-0.10 in control patients, 7.14+/-0.13 in those with Fisher grade 1 or 2 hemorrhage, and 6.95+/-0.18 in those with with Fisher grade 3 hemorrhage (P<0.05). At a 9% end-tidal desflurane level, brain tissue oxygen pressure increased to 19+/-9 mm Hg and brain tissue pH increased to 7.11+/-0.11 in patients with Fisher grade 3 hemorrhage (P<0.05 for both increases). CONCLUSION: These results show that subarachnoid hemorrhage can significantly decrease brain tissue oxygen pressure and pH related to the severity of the bleed. Increasing the desflurane concentration to 9% increased brain tissue oxygen pressure in all patients and brain tissue pH in patients with subarachnoid hemorrhage with baseline acidosis. PMID- 10691232 TI - Efficacy of two methods for reducing postbypass afterdrop. AB - BACKGROUND: Afterdrop, defined as the precipitous reduction in core temperature after cardiopulmonary bypass, results from redistribution of body heat to inadequately warmed peripheral tissues. The authors tested two methods of ameliorating afterdrop: (1) forced-air warming of peripheral tissues and (2) nitroprusside-induced vasodilation. METHODS: Patients were cooled during cardiopulmonary bypass to approximately 32 degrees C and subsequently rewarmed to a nasopharyngeal temperature near 37 degrees C and a rectal temperature near 36 degrees C. Patients in the forced-air protocol (n = 20) were assigned randomly to forced-air warming or passive insulation on the legs. Active heating started with rewarming while undergoing bypass and was continued for the remainder of surgery. Patients in the nitroprusside protocol (n = 30) were assigned randomly to either a control group or sodium nitroprusside administration. Pump flow during rewarming was maintained at 2.5 l x m(-2) x min(-1) in the control patients and at 3.0 l x m(-2) x min(-1) in those assigned to sodium nitroprusside. Sodium nitroprusside was titrated to maintain a mean arterial pressure near 60 mm Hg. In all cases, a nasopharyngeal probe evaluated core (trunk and head) temperature and heat content. Peripheral compartment (arm and leg) temperature and heat content were estimated using fourth-order regressions and integration over volume from 18 intramuscular needle thermocouples, nine skin temperatures, and "deep" hand and foot temperature. RESULTS: In patients warmed with forced air, peripheral tissue temperature was higher at the end of warming and remained higher until the end of surgery. The core temperature afterdrop was reduced from 1.2+/-0.2 degrees C to 0.5+/-0.2 degrees C by forced-air warming. The duration of afterdrop also was reduced, from 50+/-11 to 27+/-14 min. In the nitroprusside group, a rectal temperature of 36 degrees C was reached after 30+/-7 min of rewarming. This was only slightly faster than the 40+/-13 min necessary in the control group. The afterdrop was 0.8+/-0.3 degrees C with nitroprusside and lasted 34+/-10 min which was similar to the 1.1+/-0.3 degrees C afterdrop that lasted 44+/-13 min in the control group. CONCLUSIONS: Cutaneous warming reduced the core temperature afterdrop by 60%. However, heat-balance data indicate that this reduction resulted primarily because forced-air heating prevented the typical decrease in body heat content after discontinuation of bypass, rather than by reducing redistribution. Nitroprusside administration slightly increased peripheral tissue temperature and heat content at the end of rewarming. However, the core-to peripheral temperature gradient was low in both groups. Consequently, there was little redistribution in either case. PMID- 10691233 TI - Relative importance of flow versus pressure in splanchnic perfusion during cardiopulmonary bypass in rabbits. AB - BACKGROUND: Decreased gastrointestinal perfusion has been reported during cardiopulmonary bypass (CPB). Conflicting results have been published concerning thresholds of pressure and flow to avoid splanchnic ischemia during CPB. This study compared splanchnic perfusion during independent and randomized variations of CPB pump flow or arterial pressure. METHODS: Ten rabbits were studied during mild hypothermic (36 degrees C) nonpulsatile CPB using neonatal oxygenators. Simultaneous measurements of tissue blood flow in four different splanchnic areas (gastric, jejunum, ileum, and liver) were performed by laser Doppler flowmetry (LDF) before CPB (T0) and during a 4-step factorial experimental block design. Pressure and flow were alternatively high or low in random order. RESULTS: Laser Doppler flowmetry was significantly lower than pre-CPB value but was better preserved (analysis of covariance) in all organs, except liver, when CPB flow was high, whatever the pressure. Splanchnic LDF values in the low- versus high-flow groups expressed as perfusion unit were (mean +/- SD): stomach, 94+/-66 versus 137+/-75; jejunum, 118+/-78 versus 172+/-75; ileum, 95+/-72 versus 146+/-83; and liver, 79+/-72 versus 108+/-118. No significant difference of LDF was observed between the high- and low-pressure groups, whatever the flow, except for liver: stomach, 115+/-64 versus 117+/-83; jejunum, 141+/-80 versus 148+/-83; ileum, 127+/-87 versus 114+/-76; liver, 114+/-88 versus 73+/-70. CONCLUSION: Prevention of splanchnic ischemia during CPB should focus on preservation of high CPB blood flow rather than on high pressure. PMID- 10691234 TI - Long-lasting hyperalgesia induced by fentanyl in rats: preventive effect of ketamine. AB - BACKGROUND: It has been reported that mu-opioid receptor activation leads to a sustained increase in glutamate synaptic effectiveness at the N-methyl-D aspartate (NMDA) receptor level, a system associated with central hypersensitivity to pain. One hypothesis is that postoperative pain may result partly from the activation of NMDA pain facilitatory processes induced by opiate treatment per se. The authors tested here the effectiveness of the opiate analgesic fentanyl for eliciting a delayed enhancement in pain sensitivity. METHODS: The consequences of four bolus injections (every 15 min) of fentanyl (20 100 microg/kg per injection, subcutaneously) on immediate (for several hours) and long-term (for several days) sensitivity to nociceptive stimuli in the rat (paw pressure vocalization test) were evaluated. The effects of the combination of the NMDA-receptor antagonist ketamine (10 mg/kg, subcutaneously) with fentanyl also were assessed. RESULTS: Fentanyl administration exhibited a biphasic time dependent effect: first, an early response (for 2-5 h) associated with a marked increase in nociceptive threshold (analgesia), and second, a later response associated with sustained lowering of the nociceptive threshold (5 days for the longest effect) below the basal value (30% of decrease for the maximal effect) indicative of hyperalgesia. The higher the fentanyl dose used, the more pronounced was the fentanyl-induced hyperalgesia. Ketamine pretreatment, which had no analgesic effect on its own, enhanced the earlier response (analgesia) and prevented the development of long-lasting hyperalgesia. CONCLUSIONS: Fentanyl activates NMDA pain facilitatory processes, which oppose analgesia and lead to long-lasting enhancement in pain sensitivity. PMID- 10691235 TI - Norepinephrine facilitates inhibitory transmission in substantia gelatinosa of adult rat spinal cord (part 1): effects on axon terminals of GABAergic and glycinergic neurons. AB - BACKGROUND: The activation of descending norepinephrine-containing fibers from the brain stem inhibits nociceptive transmission at the spinal level. How these descending noradrenergic pathways exert the analgesic effect is not understood fully. Membrane hyperpolarization of substantia gelatinosa (Rexed lamina II) neurons by the activation of alpha2 receptors may account for depression of pain transmission. In addition, it is possible that norepinephrine affects transmitter release in the substantia gelatinosa. METHODS: Adult male Sprague-Dawley rats (9 10 weeks of age, 250-300 g) were used in this study. Transverse spinal cord slices were cut from the isolated lumbar cord. The blind whole-cell patch-clamp technique was used to record from neurons. The effects of norepinephrine on the frequency and amplitude of miniature excitatory and inhibitory postsynaptic currents were evaluated. RESULTS: In the majority of substantia gelatinosa neurons tested, norepinephrine (10-100 microM) dose-dependently increased the frequency of gamma-aminobutyric acid (GABA)ergic and glycinergic miniature inhibitory postsynaptic currents; miniature excitatory postsynaptic currents were unaffected. This augmentation was mimicked by an alpha1-receptor agonist, phenylephrine (10-60 microM), and inhibited by alpha1-receptor antagonists prazosin (0.5 microM) and 2-(2,6-dimethoxyphenoxyethyl) amino-methyl-1,4 benzodioxane (0.5 microM). Neither postsynaptic responsiveness to exogenously applied GABA and glycine nor the kinetics of GABAergic and glycinergic inhibitory postsynaptic currents were affected by norepinephrine. CONCLUSION: These results suggest that norepinephrine enhances inhibitory synaptic transmission in the substantia gelatinosa through activation of presynaptic alpha1 receptors, thus providing a mechanism underlying the clinical use of alpha1 agonists with local anesthetics in spinal anesthesia. PMID- 10691236 TI - Norepinephrine facilitates inhibitory transmission in substantia gelatinosa of adult rat spinal cord (part 2): effects on somatodendritic sites of GABAergic neurons. AB - BACKGROUND: It has been reported previously that norepinephrine, when applied to the spinal cord dorsal horn, excites a subpopulation of dorsal horn neurons, presumably inhibitory interneurons. In the current study, the authors tested whether norepinephrine could activate inhibitory interneurons, specifically those that are "GABAergic." METHODS: A transverse slice was obtained from a segment of the lumbar spinal cord isolated from adult male Sprague-Dawley rats. Whole-cell patch-clamp recordings were made from substantia gelatinosa neurons using the blind patch-clamp technique. The effects of norepinephrine on spontaneous GABAergic inhibitory postsynaptic currents were studied. RESULTS: In the majority of substantia gelatinosa neurons tested, norepinephrine (10-60 microM) significantly increased both the frequency and the amplitude of GABAergic inhibitory postsynaptic currents. These increases were blocked by tetrodotoxin (1 microM). The effects of norepinephrine were mimicked by the alpha1-receptor agonist phenylephrine (10-80 microM) and inhibited by the alpha1-receptor antagonist WB-4101 (0.5 microM). Primary-afferent-evoked polysynaptic excitatory postsynaptic potentials or excitatory postsynaptic currents in wide-dynamic-range neurons of the deep dorsal horn were also attenuated by phenylephrine (40 microM). CONCLUSION: The observations suggest that GABAergic interneurons possess somatodendritic alpha1 receptors, and activation of these receptors excites inhibitory interneurons. The alpha1 actions reported herein may contribute to the analgesic action of intrathecally administered phenylephrine. PMID- 10691237 TI - Intrathecally administered cGMP-dependent protein kinase Ialpha inhibitor significantly reduced the threshold for isoflurane anesthesia: implication for a novel role of cGMP-dependent protein kinase Ialpha. AB - BACKGROUND: Inhalational anesthetics have been shown to inhibit the nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway. Previous studies indicated that inhibition of the NO-cGMP pathway decreased the level of consciousness and augmented anesthesia, analgesia, or sedation. The current study investigated the possible involvement of cGMP-dependent protein kinases (PKGs) as major effectors for the NO-cGMP pathway in the anesthetic state. METHODS: After initial baseline determination of the minimum alveolar concentration (MAC), a selective cGMP dependent protein kinase Ialpha inhibitor, Rp-8-p-CPT-cGMPS, or an NO donor, (NOC 12), were injected intrathecally. Ten minutes later, MAC measurement was repeated. The rats also were evaluated for the presence of locomotor dysfunction by intrathecal administration of Rp-8-p-CPT-cGMPS and NOC-12 in conscious rats. RESULTS: Rp-8-p-CPT-cGMPS at 25, 50, 100, and 200 microg/10 microl produced a significant decrease from isoflurane control MAC of -4+/-3.1%, 16+/-4.5%, 30+/ 5.0%, and 21+/-2.2%, respectively, which was not accompanied by significant changes in either blood pressure or heart rate. In contrast, NOC-12 at 100 microg/10 microl caused an increase from isoflurane control MAC of 23+/-5.8%, which was accompanied by significant decrease in blood pressure but not in heart rate. Rp-8-p-CPT-cGMPS (100 microg/10 microl) produced a significant reversal of isoflurane MAC increase induced by NOC-12 (100 microg/10 microl), which was accompanied by significant reversal of the reduction of blood pressure induced by NOC-12. Locomotor activity was not changed. CONCLUSIONS: The results indicate that cGMP-dependent protein kinase Ialpha inhibitor not only markedly reduces MAC for isoflurane, but also completely blocks the NO-induced increase in isoflurane MAC, which suggests that cGMP-dependent protein kinase Ialpha may mediate the action for the NO-cGMP pathway in anesthetic mechanisms at the spinal cord level. PMID- 10691238 TI - Synergistic effect between intrathecal non-NMDA antagonist and gabapentin on allodynia induced by spinal nerve ligation in rats. AB - BACKGROUND: Glutamate and non-N-methyl-D-aspartate (NMDA) receptors have been implicated in the development of neuroplasticity in the spinal cord in neuropathic pain. The spinal cord has been identified as one of the sites of the analgesic action of gabapentin. In the current study, the authors determined the antiallodynic effect of intrathecal 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) in a rat model of neuropathic pain. Also tested was a hypothesis that intrathecal injection of CNQX and gabapentin produces a synergistic effect on allodynia in neuropathic rats. METHODS: Allodynia was produced in rats by ligation of the left L5 and L6 spinal nerves. Allodynia was determined by application of von Frey filaments to the left hind paw. Through an implanted intrathecal catheter, 10-100 microg gabapentin or 0.5-8 microg CNQX disodium (a water-soluble formulation of CNQX) was injected in conscious rats. Isobolographic analysis was performed comparing the interaction of intrathecal gabapentin and CNQX using the ED50 dose ratio of 15:1. RESULTS: Intrathecal treatment with gabapentin or CNQX produced a dose-dependent increase in the withdrawal threshold to mechanical stimulation. The ED50 for gabapentin and CNQX was 45.9+/-4.65 and 3.4+/-0.22 microg, respectively. Intrathecal injection of a combination of CNQX and gabapentin produced a strong synergistic antiallodynic effect in neuropathic rats. CONCLUSIONS: This study shows that intrathecal administration of CNQX exhibits an antiallodynic effect in this rat model of neuropathic pain. Furthermore, CNQX and gabapentin, when combined intrathecally, produce a potent synergistic antiallodynic effect on neuropathic pain in spinal nerve-ligated rats. PMID- 10691239 TI - Actions of midazolam on GABAergic transmission in substantia gelatinosa neurons of adult rat spinal cord slices. AB - BACKGROUND: Although intrathecal administration of midazolam has been found to produce analgesia, how midazolam exerts this effect is not understood fully at the neuronal level in the spinal cord. METHODS: The effects of midazolam on either electrically evoked or spontaneous inhibitory transmission and on a response to exogenous gamma-aminobutyric acid (GABA), a GABA(A)-receptor agonist, muscimol, or glycine were evaluated in substantia gelatinosa neurons of adult rat spinal cord slices by using the whole-cell patch-clamp technique. RESULTS: Bath applied midazolam (1 microM) prolonged the decay phase of evoked and miniature inhibitory postsynaptic currents (IPSCs), mediated by GABA(A) receptors, without a change in amplitudes, while not affecting glycine receptor-mediated miniature inhibitory postsynaptic currents in both the decay phase and the amplitude. Either GABA- or muscimol-induced currents were enhanced in amplitude by midazolam (0.1 microM) in a manner sensitive to a benzodiazepine receptor antagonist, flumazenil (1 microM); glycine currents were, however, unaltered by midazolam. CONCLUSIONS: Midazolam augmented both the duration of GABA-mediated synaptic current and the amplitude of GABA-induced current by acting on the GABA(A) benzodiazepine receptor in substantia gelatinosa neurons; this would increase the inhibitory GABAergic transmission. This may be a possible mechanism for antinociception by midazolam. PMID- 10691240 TI - Xenon does not alter cardiac function or major cation currents in isolated guinea pig hearts or myocytes. AB - BACKGROUND: The noble gas xenon (Xe) has been used as an inhalational anesthetic agent in clinical trials with little or no physiologic side effects. Like nitrous oxide, Xe is believed to exert minimal unwanted cardiovascular effects, and like nitrous oxide, the vapor concentration to achieve 1 minimum alveolar concentration (MAC) for Xe in humans is high, i.e., 70-80%. In the current study, concentrations of up to 80% Xe were examined for possible myocardial effects in isolated, erythrocyte-perfused guinea pig hearts and for possible effects on altering major cation currents in isolated guinea pig cardiomyocytes. METHODS: Isolated guinea pigs hearts were perfused at 70 mm Hg via the Langendorff technique initially with a salt solution at 37 degrees C. Hearts were then perfused with fresh filtered (40-microm pore) and washed canine erythrocytes diluted in the salt solution equilibrated with 20% O2 in nitrogen (control), with 20% O2, 40% Xe, and 40% N2, (0.5 MAC), or with 20% O2 and 80% Xe (1 MAC), respectively. Hearts were perfused with 80% Xe for 15 min, and bradykinin was injected into the blood perfusate to test endothelium-dependent vasodilatory responses. Using the whole-cell patch-clamp technique, 80% Xe was tested for effects on the cardiac ion currents, the Na+, the L-type Ca2+, and the inward rectifier K+ channel, in guinea pig myocytes suffused with a salt solution equilibrated with the same combinations of Xe, oxygen, and nitrogen as above. RESULTS: In isolated hearts, heart rate, atrioventricular conduction time, left ventricular pressure, coronary flow, oxygen extraction, oxygen consumption, cardiac efficiency, and flow responses to bradykinin were not significantly (repeated measures analysis of variance, P>0.05) altered by 40% or 80% Xe compared with controls. In isolated cardiomyocytes, the amplitudes of the Na+, the L-type Ca2+, and the inward-rectifier K+ channel over a range of voltages also were not altered by 80% Xe compared with controls. CONCLUSIONS: Unlike hydrocarbon-based gaseous anesthetics, Xe does not significantly alter any measured electrical, mechanical, or metabolic factors, or the nitric oxide dependent flow response in isolated hearts, at least partly because Xe does not alter the major cation currents as shown here for cardiac myocytes. The authors' results indicate that Xe, at approximately 1 MAC for humans, has no physiologically important effects on the guinea pig heart. PMID- 10691241 TI - Bupivacaine inhibits acylcarnitine exchange in cardiac mitochondria. AB - BACKGROUND: The authors previously reported that secondary carnitine deficiency may sensitize the heart to bupivacaine-induced arrhythmias. In this study, the authors tested whether bupivacaine inhibits carnitine metabolism in cardiac mitochondria. METHODS: Rat cardiac interfibrillar mitochondria were prepared using a differential centrifugation technique. Rates of adenosine diphosphate stimulated (state III) and adenosine diphosphate-limited (state IV) oxygen consumption were measured using a Clark electrode, using lipid or nonlipid substrates with varying concentrations of a local anesthetic. RESULTS: State III respiration supported by the nonlipid substrate pyruvate (plus malate) is minimally affected by bupivacaine concentrations up to 2 mM. Lower concentrations of bupivacaine inhibited respiration when the available substrates were palmitoylcarnitine or acetylcarnitine; bupivacaine concentration causing 50% reduction in respiration (IC50 +/- SD) was 0.78+/-0.17 mM and 0.37+/-0.03 mM for palmitoylcarnitine and acetylcarnitine, respectively. Respiration was equally inhibited by bupivacaine when the substrates were palmitoylcarnitine alone, or palmitoyl-CoA plus carnitine. Bupivacaine (IC50 = 0.26+/-0.06 mM) and etidocaine (IC50 = 0.30+/-0.12 mM) inhibit carnitine-stimulated pyruvate oxidation similarly, whereas the lidocaine IC50 is greater by a factor of roughly 5, (IC50 = 1.4+/-0.26 mM), and ropivacaine is intermediate, IC50 = 0.5+/-0.28 mM. CONCLUSIONS: Bupivacaine inhibits mitochondrial state III respiration when acylcarnitines are the available substrate. The substrate specificity of this effect rules out bupivacaine inhibition of carnitine palmitoyl transferases I and II, carnitine acetyltransferase, and fatty acid beta-oxidation. The authors hypothesize that differential inhibition of carnitine-stimulated pyruvate oxidation by various local anesthetics supports the clinical relevance of inhibition of carnitine-acylcarnitine translocase by local anesthetics with a cardiotoxic profile. PMID- 10691242 TI - Differential effects of anesthetic and nonanesthetic cyclobutanes on neuronal voltage-gated sodium channels. AB - BACKGROUND: Despite their key role in the generation and propagation of action potentials in excitable cells, voltage-gated sodium (Na+) channels have been considered to be insensitive to general anesthetics. The authors tested the sensitivity of neuronal Na+ channels to structurally similar anesthetic (1-chloro 1,2,2-trifluorocyclobutane; F3) and nonanesthetic (1,2 dichlorohexafluorocyclobutane; F6) polyhalogenated cyclobutanes by neurochemical and electrophysiologic methods. METHODS: Synaptosomes (pinched-off nerve terminals) from adult rat cerebral cortex were used to determine the effects of F3 and F6 on 4-aminopyridine- or veratridine-evoked (Na+ channel-dependent) glutamate release (using an enzyme-coupled spectrofluorimetric assay) and increases in intracellular Ca2+ ([Ca2+]i) (using ion-specific spectrofluorimetry). Effects of F3 and F6 on Na+ currents were evaluated directly in rat lumbar dorsal root ganglion neurons by whole-cell patch-clamp recording. RESULTS: F3 inhibited glutamate release evoked by 4-aminopyridine (inhibitory concentration of 50% [IC50] = 0.77 mM [approximately 0.8 minimum alveolar concentration (MAC)] or veratridine (IC50 = 0.42 mM [approximately 0.4 MAC]), and veratridine-evoked increases in [Ca2+]i (IC50 = 0.5 mM [approximately 0.5 MAC]) in synaptosomes; F6 had no significant effects up to 0.05 mM (approximately twice the predicted MAC). F3 caused reversible membrane potential-independent inhibition of peak Na+ currents (70+/-9% block at 0.6 mM [approximately 0.6 MAC]), and a hyperpolarizing shift in the voltage-dependence of steady state inactivation in dorsal root ganglion neurons (-21+/-9.3 mV at 0.6 mM). F6 inhibited peak Na+ currents to a lesser extent (16+/-2% block at 0.018 mM [predicted MAC]) and had minimal effects on steady state inactivation. CONCLUSIONS: The anesthetic cyclobutane F3 significantly inhibited Na+ channel mediated glutamate release and increases in [Ca2+]i. In contrast, the nonanesthetic cyclobutane F6 had no significant effects at predicted anesthetic concentrations. F3 inhibited dorsal root ganglion neuron Na+ channels with a potency and by mechanisms similar to those of conventional volatile anesthetics; F6 was less effective and did not produce voltage-dependent block. This concordance between anesthetic activity and Na+ channel inhibition supports a role for presynaptic Na+ channels as targets for general anesthetic effects and suggests that shifting the voltage-dependence of Na+ channel inactivation is an important property of volatile anesthetic compounds. PMID- 10691243 TI - Propofol-induced modifications of cardiomyocyte calcium transient and sarcoplasmic reticulum function in rats. AB - BACKGROUND: Propofol is considered to be an anesthetic agent with few or no negative inotropic effects. This study evaluated a possible direct depressant effect of propofol on sarcoplasmic reticulum Ca2+ accumulation and cardiomyocytes. METHODS: The effects of propofol on intracellular Ca2+ transients were evaluated in isolated rat cardiomyocytes using a microfluorometric technique with Indo-1. Sarcoplasmic reticulum function was also assessed by measuring the oxalate-stimulated Ca2+ uptake from homogenates of rat ventricles. RESULTS: The Ca2+ uptake capacity of the sarcoplasmic reticulum was decreased by propofol (10( 4) M). Large concentrations of propofol decreased the rate of decrease of the intracellular Ca2+ transient, which resulted in an increase of diastolic Ca2+ when the diastolic interval was decreased. The increased diastolic Ca2+ also resulted in a decrease in Ca2+ transient. This effect appeared for lower doses (10(-5) M) after a short diastolic pause rather than after a long (2- to 3-min) rest (appearing at 10(-4) M). CONCLUSIONS: For doses more than 10(-5) M, propofol induces a Ca2+ uptake capacity impairment of the sarcoplasmic reticulum. This is probably responsible for a slowing of the decrease of the Ca2+ transient, which in turn increases the diastolic Ca2+ for high heart rate. These diastolic modifications may participate in the slight negative inotropic effect of the drug. PMID- 10691244 TI - Primate pleuroesophageal tissue barrier frequency response and esophageal pressure waveform bandwidth in health and acute lung injury. AB - BACKGROUND: Dynamic intraesophageal pressure (Pes) is used to estimate intrapleural pressure (Ppl) to calculate lung compliance and resistance. This study investigated the nonhuman primate Ppl-Pes tissue barrier frequency response and the dynamic response requirements of Pes manometers. METHODS: In healthy monkeys and monkeys with acute lung injury undergoing ventilation, simultaneous Ppl and Pes were measured directly to determine the Ppl-Pes tissue barrier amplitude frequency response, using the swept-sine wave technique. The bandwidths of physiologic Pes waveforms acquired during conventional mechanical ventilation were calculated using digital low-pass signal filtering. RESULTS: The Ppl-Pes tissue barrier is amplitude-uniform within the bandwidth of conventional Pes waveforms in healthy and acute lung injury lungs, and does not significantly attenuate Ppl-Pes signal transmission between 1 and 40 Hz. At Pes frequencies higher than conventional clinical regions of interest the Ppl-Pes barrier resonates significantly, is pressure amplitude dependent at low-pressure offsets, and is significantly altered by acute lung injury. Allowing for 5% or less Pes waveform error, the maximum Pes bandwidths during conventional ventilation were 1.9 Hz and 3.4 Hz for physiologic and extreme-case waveforms in healthy lungs and 4.6 Hz and 8.5 Hz during acute lung injury. CONCLUSIONS: In monkeys, the Ppl-Pes tissue barrier has a frequency response suitable for Ppl estimation during low frequency mechanical ventilation, and Pes manometers should have a minimum uniform frequency response up to 8.5 Hz. However, the Ppl-Pes tissue barrier adversely affects the accurate estimation of dynamic Ppl at high frequencies, with varied airway pressure amplitudes and offsets, such as the Ppl encountered during high-frequency oscillatory ventilation. PMID- 10691245 TI - Isoflurane action in the spinal cord blunts electroencephalographic and thalamic reticular formation responses to noxious stimulation in goats. AB - BACKGROUND: Isoflurane depresses the electroencephalographic (EEG) activity and exerts part of its anesthetic effect in the spinal cord. The authors hypothesized that isoflurane would indirectly depress the EEG and subcortical response to noxious stimulation in part by a spinal cord action. METHODS: Depth electrodes were inserted into the midbrain reticular formation (MRF) and thalamus of six of seven isoflurane-anesthetized goats, and needle-electrodes were placed into the skull periosteum. In five of seven goats, an MRF microelectrode recorded single unit activity. The jugular veins and carotid arteries were isolated to permit cranial bypass and differential isoflurane delivery. A noxious mechanical stimulus (1 min) was applied to a forelimb dewclaw at each of two cranial-torso isoflurane combinations: 1.1+/-0.3%-1.2+/-0.3% and 1.1+/-0.3-0.3+/-0.1% (mean +/- SD). RESULTS: When cranial-torso isoflurane was 1.1-1.2%, the noxious stimulus did not alter the EEG. When torso isoflurane was decreased to 0.3%, the noxious stimulus activated the MRF, thalamic, and bifrontal-hemispheric regions (decreased high-amplitude, low-frequency power). For all channels combined, total (-33+/-15%), delta(-51+/-22%), theta (-33+/-19%), and alpha (-26+/-16%) power decreased after the noxious stimulus (P<0.05); beta power was unchanged. The MRF unit responses to the noxious stimulus were significantly higher when the spinal cord isoflurane concentration was 0.3% (1,286+/-1,317 impulses/min) as compared with 1.2% (489+/-437 impulses/min, P<0.05). CONCLUSIONS: Isoflurane blunted the EEG and MRF-thalamic response to noxious stimulation in part via an action in the spinal cord. PMID- 10691246 TI - Midazolam selectively potentiates the A(2A) - but not A1- receptor--mediated effects of adenosine: role of nucleoside transport inhibition and clinical implications. AB - BACKGROUND: Inhibition of adenosine metabolism offers a unique approach to harness the cardioprotective properties of adenosine in a site- and event specific manner. Benzodiazepines inhibit adenosine metabolism by blocking nucleoside transporter. Therefore, the authors studied the binding affinities of structurally different benzodiazepines to nucleoside transporter and benzodiazepine-induced potentiation of A1-adenosine (negative dromotropy) and A2A adenosine (coronary vasodilation) receptor-mediated effects. METHODS: In membranes from porcine striatum and guinea pig ventricle, competition binding assays to displace [3H]nitrobenzylmercaptopurine riboside ([3H]NBMPR) from nucleoside transporter were performed using alprazolam, chlorodiazepoxide, diazepam, flurazepam, and midazolam. The augmentation by the most potent benzodiazepine of A1- and A2A-adenosine receptor-mediated responses, elicited by exogenous administration of adenosine or brief periods of global hypoxia, was subsequently studied in guinea pig Langendorff-perfused hearts. RESULTS: All benzodiazepines completely displaced [3H]NBMPR in a concentration-dependent manner with Hill coefficients not significantly different from unity in both striatal and ventricular membranes. Midazolam was the most potent inhibitor of nucleoside transporter (ventricle:pKi = 5.22+/-0.41, Ki = 6 microM). In isolated hearts, midazolam (5, 10, 20 microM) significantly augmented coronary flow in a concentration-dependent manner in the presence of adenosine (30 nM), an effect reversed by ZM 241385, a selective A2A-receptor antagonist. In contrast, midazolam did not increase the effect of adenosine (30 nM) on atrioventricular conduction. Similarly, midazolam potentiated A2A- but not A1-receptor-mediated effects of endogenous adenosine released during hypoxia. CONCLUSIONS: Structurally distinct benzodiazepines inhibit nucleoside transporter to different degrees. Midazolam selectively augments A2A- but not A1-receptor-mediated effects of adenosine by inhibiting nucleoside transporter. PMID- 10691247 TI - Perioperative heat balance. AB - Hypothermia during general anesthesia develops with a characteristic three-phase pattern. The initial rapid reduction in core temperature after induction of anesthesia results from an internal redistribution of body heat. Redistribution results because anesthetics inhibit the tonic vasoconstriction that normally maintains a large core-to-peripheral temperature gradient. Core temperature then decreases linearly at a rate determined by the difference between heat loss and production. However, when surgical patients become sufficiently hypothermic, they again trigger thermoregulatory vasoconstriction, which restricts core-to peripheral flow of heat. Constraint of metabolic heat, in turn, maintains a core temperature plateau (despite continued systemic heat loss) and eventually reestablishes the normal core-to-peripheral temperature gradient. Together, these mechanisms indicate that alterations in the distribution of body heat contribute more to changes in core temperature than to systemic heat imbalance in most patients. Just as with general anesthesia, redistribution of body heat is the major initial cause of hypothermia in patients administered spinal or epidural anesthesia. However, redistribution during neuraxial anesthesia is typically restricted to the legs. Consequently, redistribution decreases core temperature about half as much during major conduction anesthesia. As during general anesthesia, core temperature subsequently decreases linearly at a rate determined by the inequality between heat loss and production. The major difference, however, is that the linear hypothermia phase is not discontinued by reemergence of thermoregulatory vasoconstriction because constriction in the legs is blocked peripherally. As a result, in patients undergoing large operations with neuraxial anesthesia, there is the potential of development of serious hypothermia. Hypothermic cardiopulmonary bypass is associated with enormous changes in body heat content. Furthermore, rapid cooling and rewarming produces large core-to peripheral, longitudinal, and radial tissue temperature gradients. Inadequate rewarming of peripheral tissues typically produces a considerable core-to peripheral gradient at the end of bypass. Subsequently, redistribution of heat from the core to the cooler arms and legs produces an afterdrop. Afterdrop magnitude can be reduced by prolonging rewarming, pharmacologic vasodilation, or peripheral warming. Postoperative return to normothermia occurs when brain anesthetic concentration decreases sufficiently to again trigger normal thermoregulatory defenses. However, residual anesthesia and opioids given for treatment of postoperative pain decreases the effectiveness of these responses. Consequently, return to normothermia often needs 2-5 h, depending on the degree of hypothermia and the age of the patient. PMID- 10691248 TI - Awareness during anesthesia. PMID- 10691249 TI - Erroneous conclusion from processed electroencephalogram with changing anesthetic depth. PMID- 10691251 TI - Angina as an indication for noncardiac surgery: the case of the coronary subclavian steal syndrome. PMID- 10691250 TI - Uneventful propofol anesthesia in a patient with coexisting hereditary coproporphyria and hereditary angioneurotic edema. PMID- 10691252 TI - Ulnar neuropathy in medical patients. PMID- 10691253 TI - Reversible catecholamine-induced cardiomyopathy by subcutaneous injections of epinephrine solution in an anesthetized patient. PMID- 10691254 TI - Pharyngeal mucosa pressures. PMID- 10691255 TI - Spinal anesthesia in preeclamptic patients--"supportive" evidence. PMID- 10691256 TI - Saline infusion, acidosis, and the Stewart approach. PMID- 10691257 TI - Article supports findings of previous comparison. PMID- 10691258 TI - Avoiding latrogenic hyperchloremic acidosis--call for a new crystalloid fluid. PMID- 10691259 TI - Dynamic response to volatile anesthetics has been examined before. PMID- 10691260 TI - Chemical skinning artifact appears to increase sensitivity of masseter muscle to halothane and succinylcholine. PMID- 10691261 TI - Fatal hydrocephalus in a patient with neurofibromatosis. PMID- 10691262 TI - Security system for transducer holders. PMID- 10691263 TI - An easy formula to remember the laryngeal mask airway size-patient weight relationship. PMID- 10691264 TI - Paraplegia after cystectomy and epidural anesthesia. PMID- 10691265 TI - Extrahepatic biliary obstruction: magnetic resonance imaging compared with endoscopic ultrasonography. AB - BACKGROUND AND STUDY AIMS: The aim of this study was to compare prospectively the diagnostic efficacy of magnetic resonance (MR) imaging and endoscopic ultrasonography (EUS) in extrahepatic biliary obstruction. PATIENTS AND METHODS: A total of 50 patients with suspected benign or malignant extrahepatic biliary obstruction underwent MR imaging, including MR cholangiopancreatography, and EUS, within a median time delay of 1 day. The final diagnosis was established by endoscopic retrograde cholangiopancreatography in 37 cases, intraoperative cholangiography in nine cases, and clinical and biochemical follow-up in four cases. RESULTS: In total, 33 patients had extrahepatic biliary obstruction, of benign origin in 21 cases and of malignant origin in 12 cases, whereas 17 had no evidence of obstruction. The sensitivity and specificity of MR imaging were 91% and 94 %, respectively. There were one false-positive and three false-negative results, all related to choledochal sludge. The corresponding values for EUS were 97% and 88%. There were two false-positive results and one false-negative result. False-positive diagnoses were related to the presumed presence of biliary sludge and choledocholithiasis, whereas the false-negative diagnosis occurred in one patient with a final diagnosis of sludge. No significant difference in sensitivity and specificity was observed between the two imaging methods (P>0.05). CONCLUSION: In our study MR imaging was as accurate as EUS in the diagnosis of extrahepatic biliary obstruction. PMID- 10691266 TI - Analysis of the risk factors associated with endoscopic sphincterotomy techniques: preliminary results of a prospective study, with emphasis on the reduced risk of acute pancreatitis with low-dose anticoagulation treatment. AB - BACKGROUND AND STUDY AIMS: The aim of the present study was to analyze the risk factors associated with complications of endoscopic sphincterotomy (ES). PATIENTS AND METHODS: In all consecutive endoscopic sphincterotomies carried out between September 1994 and December 1996, the possible risk factors (12 patient-related factors and 12 procedure-related ones), as well as the concomitant medical treatment, indications, techniques, and success of endoscopic sphincterotomy were evaluated prospectively. Risk factors were analyzed on an exploratory basis using univariate methods. "Potential risk factors" (univariate, P<0.1) underwent multivariate analysis to determine independent "risk factors" (multivariate, P<0.05). In addition, the complication rate was calculated according to the number of potential risk factors present. RESULTS: A total of 438 patients who underwent ES were analyzed. Complications occurred in 7.5% (n = 33; acute pancreatitis 4.3%, hemorrhage 2.3 %, cholangitis 0.9%, technical 0.2%). Statistical analysis of the complications identified three independent risk factors (coagulopathy, patient age (< or =60 years, pancreas divisum), and one protective factor (pancreatic duct obstruction). The frequency of acute pancreatitis was increased by two independent risk factors (pancreas divisum, ES frequency <40 procedures/year) and was reduced if low-dose anticoagulation (unfractionated heparin or low molecular weight heparin) was administered (0.9%, one of 115 vs. 5.8%, 18 of 313; P<0.05). The effect of anticoagulation was not confounded by the presence or absence of other potential risk factors for acute pancreatitis. Neither the risk nor the severity of hemorrhage were increased by low-dose anticoagulation. Due to the low number of events, only potential risk factors for hemorrhage were identified (coagulopathy, intensive-care treatment). The overall complication rate and the incidence of pancreatitis and hemorrhage increased significantly depending on the number of simultaneous potential risk factors present (P<0.0001). CONCLUSIONS: Patients at risk for complications after endoscopic sphincterotomy can be identified by risk factor analysis. These data suggest the hypothesis that low-dose anticoagulation prior to endoscopic sphincterotomy reduces the risk of acute pancreatitis after sphincterotomy. PMID- 10691267 TI - Video manometry of the sphincter of Oddi: a new aid for interpreting manometric tracings and excluding manometric artefacts. AB - BACKGROUND AND STUDY AIMS: Endoscopic sphincter of Oddi manometry (ESOM) allows direct assessment of motor function in the sphincter of Oddi. However, variations in examination conditions and duodenal motility may have a critical effect on the results of ESOM. The aim of the present study was to develop a new method sphincter of Oddi video manometry-based on simultaneous ESOM and real-time endoscopic image analysis, and to investigate the usefulness of video manometry for detecting manometric artefacts during ESOM. PATIENTS AND METHODS: Seven consecutive patients who had undergone cholecystectomy and were referred with a suspicion of sphincter of Oddi dysfunction were investigated. Sphincter of Oddi pressure and endoscopic images (20 frames/s) were recorded simultaneously on a Synectics PC Polygraf computer system with a time-correlated basis, and then compared. RESULTS: On ESOM, 69 sphincter of Oddi phasic contractions were identified, with an average amplitude of 153.9+/-85.0 mm Hg and a duration of 7.9+/-1.2 seconds. Visual analysis of the real-time endoscopic images, replayed in cine loop by the computer, revealed 236 separate duodenal contractions, with an average frequency of 3.5+/-2.4/min (range: 1-12/min). On the ESOM tracing, 78% of the duodenal contractions had a corresponding pressure wave with an average duration of 2.8+/-0.4 seconds and an amplitude of 71.9+/-16.7 mm Hg. Other artefacts on the ESOM tracings, such as catheter movements, pseudocontractions, hyperventilation, or retching, were also easily recognized using simultaneous ESOM and real-time endoscopic image analysis. CONCLUSIONS: Video manometry of the sphincter of Oddi is a promising new method for improving the analysis and documentation of ESOM tracings. It has several advantages over the conventional technique, allowing visual detection of duodenal activity and enabling enhanced recognition of other manometric artefacts. PMID- 10691268 TI - Extension of squamous epithelium into the proximal stomach: a newly recognized mucosal abnormality. AB - BACKGROUND AND STUDY AIMS: The presence of squamous epithelium in the stomach has been rarely noted in the past with only sporadic case reports of squamous cell carcinoma of the stomach and a variety of other pathological processes. We report the documentation, over a period of 9 months, of squamous epithelium extending beyond the esophagogastric junction and into the proximal stomach in 16 patients who underwent upper endoscopy. PATIENTS AND METHODS: This newly recognized mucosal abnormality was systematically identified by both antegrade and retrograde views of the esophagogastric junction during routine upper endoscopy. Other associated mucosal abnormalities were also evaluated (Barrett's esophagus, erosive esophagitis, etc.). Biopsies were obtained from the distal esophagus, cardia, antrum and the squamous extension. Additionally, all patients underwent a detailed interview. RESULTS: Of the patients, 14 were Caucasian and two Hispanic; all were males, with a mean age 61.2+/-4.2. Indications for upper endoscopy included dysphagia, Barrett's esophagus surveillance, failure of antireflux treatment and anemia. Heartburn was reported by 12 patients (75%). None of the patients reported a history of corrosive injury, foreign body ingestion or surgery. A total of 12 patients had a solitary tongue of squamous cell extension, three had two tongues and in addition, two had squamous islands. Hiatal hernia was present in all patients, Barrett's esophagus in six (37.5 %), and esophageal stricture in four. CONCLUSION: Squamous cell extension into the proximal stomach is a newly recognized mucosal abnormality with presently unknown clinical significance. This mucosal abnormality may represent an esophageal mucosal response to proximal gastric injury. PMID- 10691269 TI - Cardiopulmonary complications during gastroscopy in patients with chronic respiratory failure undergoing long-term home oxygen therapy. AB - BACKGROUND AND STUDY AIMS: Gastric ulcer and hemorrhage are major complications in patients with chronic respiratory failure, but upper GI endoscopy tends to be avoided because of possible cardiopulmonary events. This study was designed to evaluate hypoxemia and subsequent cardiac complications during gastroscopic procedures in patients with chronic respiratory failure undergoing long-term home oxygen therapy (LHOT). PATIENTS AND METHODS: Gastroscopy was carried out in 10 patients undergoing LHOT and 10 age-matched control subjects without pulmonary diseases. Oxygen saturation and cardiac arrhythmias before and during gastroscopy were monitored. Patients were given 10 mg intramuscular scopolamine butylbromide and local anesthesia using 100-300 mg lidocaine gel 15 minutes before the procedure. Each patient continued to receive oxygen via a nasal cannula in the same dosage as their daily use. RESULTS: Decrease in oxygen saturation during endoscopic procedure was significantly greater in patients undergoing LHOT (from 95.9+/-0.9 to 93.4+/-1.7%) compared with control subjects (from 96.7+/-0.4 to 96.2+/-0.4%). There was a significant correlation between the degree of hypoxemia and the oxygen dosage required for their daily treatment in the patients (r = 0.727, P<0.02). CONCLUSIONS: These results indicate that the degree of respiratory failure influences the degree of decrease in oxygen saturation during gastroscopy. It is suggested that use of the nasal route for oxygen supply may be one of the major causes of the hypoxemia. PMID- 10691270 TI - A blinded, randomized comparison of a novel, low-dose, triple regimen with fleet phospho-soda: a study of colon cleanliness, speed and success of colonoscopy. AB - BACKGROUND AND STUDY AIMS: A clean colon is essential for an efficient examination. The aim of this study was to compare a novel low-dose, low volume triple regimen with Fleet Phospho-soda. METHODS: A blinded, experienced colonoscopist examined 132 consecutive patients randomly allocated to receive either a triple regimen consisting of senna syrup (sennoside B), Picolax (sodium picosulphate), and Klean Prep (polyethylene glycol 3350), or Fleet Phospho-soda (sodium dihydrogen phosphate and disodium phosphate dodecahydrate). The colonoscopist recorded cleanliness according to a scoring system (1-very clean to 4-solid stools), and time taken to reach the caecum. RESULTS: In the triple regimen group (n = 81), 73% scored 1 or 2 compared with 57% in the Fleet Phospho soda group (n = 51, p = 0.037 Mann-Whitney U-test). Examination to caecum was achieved in 95% of the triple regimen group and 89% of the Fleet Phospho-soda group. Among those examined as far as the caecum, the time to reach the caecum was 11 minutes (range 5-50) in the triple regimen group compared with 16 minutes (range 5-65) in the Fleet Phospho-soda group (p = 0.08, Mann-Whitney U-test). Patient tolerability was not assessed in this study. CONCLUSIONS: This novel triple regimen produces a cleaner colon than Fleet Phospho-soda, is associated with a trend towards a quicker and more efficient colonic examination, and is also 30% cheaper per patient. PMID- 10691271 TI - Does hyperbaric oxygen enhance the effect of photodynamic therapy in patients with advanced esophageal carcinoma? A clinical pilot study. AB - BACKGROUND AND STUDY AIMS: Experimental studies have shown that the cytotoxicity of porphyrins and related substances is mediated mainly by singlet oxygen and that hypoxic cells are less affected by porphyrins and light. In a clinical pilot study we assessed the use of photodynamic therapy (PDT) under hyperbaric oxygen (HBO), compared with PDT under normobaric conditions, in patients with advanced esophageal carcinoma. PATIENTS AND METHODS: After diagnostic work-up and staging, photosensitization in all patients was carried out using hematoporphyrine derivate (HpD) (2 mg/kg bodyweight 48 hours prior to PDT). We then applied light at 630 nm (KTP-Nd: YAG laser with DYE box) at dose of 300 J/cm, delivered by a fiber with a radial light-diffusing cylinder (length 1 cm), inserted through the biopsy channel of the endoscope. Of the patients, 14 (12 with stage III cancers, and two with stage IV cancers) were treated by PDT alone, and 17 patients (15 with stage III cancers, and two with stage IV cancers) received PDT under HBO at a level of 2 absolute atmospheric pressures (ATA). Transcutaneous PO2 levels of 500-750 mm Hg under HBO, compared with transcutaneous PO2 levels of 60-75 mm Hg under normobaric conditions, were measured. RESULTS: Improvements regarding dysphagia and stenosis diameter were obtained in both treatment arms with no significant differences (P = 0.36 and 0.14, respectively). The tumor length also decreased in both groups and showed a significant difference in favour of the PDT/ HBO group (P = 0.002). Kaplan-Meier statistics showed median overall survival for the PDT group and the PDT/HBO group as 7.0 and 12 months respectively. The 12-month survival rate was 28.6% for the PDT group and 41.2% for the PDT/HBO group. Logrank test showed a difference in survival in favor of the PDT/HBO group (P = 0.059). No major treatment-related complication occurred, and the 30-day mortality rate was 0%. CONCLUSIONS: Combined PDT/HBO represents a new approach in the treatment of esophageal cancer which, in this pilot study, appears to have enhanced the efficiency of PDT. PMID- 10691272 TI - Thermoplastic stents: a new concept for endoluminal prosthesis. AB - BACKGROUND AND STUDY AIMS: Intraluminal stenting of organs with stenoses or fistulae in anatomically difficult locations (for instance cardia, pylorus, large bowel), with a tendency to kinking or increased motility, still carries a high risk of stent dislocation. In the search for a solution, we report on the use of a new thermoplastic stent in animal experiments. MATERIAL AND METHODS: The new stent consists of a plastic-coated wire mesh which can be heated electrically. Once it is warmed up to 55 C, its size and shape can be changed. After being expanded by a dilatation balloon across the stenosed area, the stent can be fitted onto the inner organ surface. This guarantees a low dislocation risk and high stability. In an animal experiment, stents were endoscopically placed in the trachea and the surgically stenosed esophagus of two dogs. The animals were observed for 3 months. RESULTS: The thermostents were implanted easily and without complications. It was possible to mold the thermostent evenly onto the intraluminal wall. No stent dislocation, bleeding or perforation was observed. Upon histologic evaluation, granulation tissue was found to be growing through the wire mesh of the stent. CONCLUSION: It was shown that the stent described here can be implanted without major problems. The greater effort of the implantation procedure, in comparison with self-expanding stents, is compensated by the special mechanical characteristics of the stent. These characteristics may permit implantation in anatomically difficult locations where up to now stenting has been impossible or inadequate. PMID- 10691273 TI - Sigmoid stiffener for decompression tube placement in colonic pseudo-obstruction. AB - BACKGROUND AND STUDY AIMS: Decompression tube placement improves outcome in colonic pseudo-obstruction (CP) which is refractory to conservative measures, especially if the decompression tube is placed proximal to the hepatic flexure. We evaluate the ability of a sigmoid stiffener to facilitate more proximal colonoscopy and decompression tube placement. PATIENTS AND METHODS: A sigmoid stiffener is used in the standard fashion during colonoscopic decompression for pseudo-obstruction. After cecal wire placement, the colonoscope is withdrawn, leaving the stiffener and wire in place. By passing through the stiffener, an over-wire decompression tube can avoid sigmoid looping. We compared proximal extent of colonoscopy, tube position, endoscopy time, and patient outcomes using a sigmoid stiffener, with a control group of patients treated previously. Patients with colonic ischemia were excluded. RESULTS: Using this technique, nine consecutive colonoscopies and decompression tube placements reached the right colon. Significantly, only three of seven control colonoscopies and two control decompression tubes did so. However, improvements in procedural time and patient outcome did not reach statistical significance. No complications occurred. CONCLUSION: The use of a sigmoid stiffener during colonic decompression allows more proximal colonoscopy and decompression tube placement, with possible clinical benefit. We do not use this technique in the setting of left colon ischemia. PMID- 10691274 TI - Development of a speech-based dialogue system for report dictation and machine control in the endoscopic laboratory. AB - BACKGROUND AND STUDY AIMS: Reporting and machine control based on speech technology can enhance work efficiency in the gastrointestinal endoscopy laboratory. MATERIALS AND METHODS: The status and activation of endoscopy laboratory equipment were described as a multivariate parameter and function system. Speech recognition, text evaluation and action definition engines were installed. Special programs were developed for the grammatical analysis of command sentences, and a rule-based expert system for the definition of machine answers. A speech backup engine provides feedback to the user. Techniques were applied based on the "Hidden Markov" model of discrete word, user-independent speech recognition and on phoneme-based speech synthesis. Speech samples were collected from three male low-tone investigators. RESULTS: The dictation module and machine control modules were incorporated in a personal computer (PC) simulation program. Altogether 100 unidentified patient records were analyzed. The sentences were grouped according to keywords, which indicate the main topics of a gastrointestinal endoscopy report. They were: "endoscope", "esophagus", "cardia", "fundus", "corpus", "antrum", "pylorus", "bulbus", and "postbulbar section", in addition to the major pathological findings: "erosion", "ulceration", and "malignancy". "Biopsy" and "diagnosis" were also included. We implemented wireless speech communication control commands for equipment including an endoscopy unit, video, monitor, printer, and PC. The recognition rate was 95%. CONCLUSIONS: Speech technology may soon become an integrated part of our daily routine in the endoscopy laboratory. A central speech and laboratory computer could be the most efficient alternative to having separate speech recognition units in all items of equipment. PMID- 10691275 TI - Interventional endoscopic ultrasonography: state of the art at the new millenium. PMID- 10691277 TI - Guidelines on cleaning and disinfection in GI endoscopy. Update 1999. The European Society of Gastrointestinal Endoscopy. PMID- 10691276 TI - A little girl with pancreatitis. PMID- 10691278 TI - Protocol for reprocessing endoscopic accessories. European Society of Gastrointestinal Endoscopy. PMID- 10691279 TI - Simple hemostatic procedure for hemorrhagic duodenal ulcer: two cases of arterial hemorrhage quickly controlled by balloon compression. AB - We utilized balloon compression in two cases of arterial hemorrhage from duodenal ulcers. The bleeding was quickly controlled in both cases. The advantages of this technique are its simplicity and ease of performance, and the fact that it does not require precise identification of the bleeding point in the duodenal bulb. No serious complications, such as perforation or stenosis, are associated with this technique. During the healing of the ulcer, balloon expansion may result in decreased duodenal bulb deformity. The following points, however, should be clarified in future studies: a) the stability of the duodenal bulb after longer term balloon compression, and b) the optimum amount of cold water to be injected into the balloon and the optimum compression time. PMID- 10691280 TI - Gut perforation caused by biliary endoprosthesis. AB - Two cases are reported of perforation of the gut caused by biliary endoprosthesies in the three-year period 1993-1995. The first patient was an 81 year-old man who had perforation of the terminal ileum caused by a straight 10 French/7 cm stent which had been dislodged from the bile duct; he underwent laparotomy but did not recover. The second patient was an 86-year-old man who had perforation of the sigmoid colon caused by a straight 7 French/5 cm stent left in the duodenum during a stent exchange procedure; he was successfully treated laparoscopically. Two cases of gut perforation in a three-year period is a rather high rate of this rare complication of placement of biliary endoprostheses. PMID- 10691281 TI - Multiple pulmonary glue emboli following gastric variceal obliteration. PMID- 10691282 TI - Laparoscopic repair of diaphragmatic iatrogenic hernia. PMID- 10691283 TI - Surgical clips incorporated into a duodenal ulcer: a rare complication after elective laparoscopic cholecystectomy. PMID- 10691284 TI - Gastroileocutaneous fistula: an unusual complication of percutaneous endoscopic gastrostomy. PMID- 10691285 TI - Experience of laparoscopic cecoplication for mobile cecum. PMID- 10691286 TI - Composite carcinoma-carcinoid tumor of the rectum treated by endoscopic removal. PMID- 10691287 TI - Esophageal aspergillosis: an unusual endoscopic finding. PMID- 10691289 TI - Par-4: an emerging pivotal player in neuronal apoptosis and neurodegenerative disorders. AB - Prostate apoptosis response-4 (Par-4) is a 38-kDa protein initially identified as the product of a gene upregulated in prostate tumor cells undergoing apoptosis. Par-4 contains both a death domain and a leucine zipper domain, and has been shown to interact with several proteins known to modulate apoptosis, including protein kinase Czeta, Bcl-2, and caspase-8. A rapid increase in Par-4 levels occurs in neurons undergoing apoptosis in a variety of paradigms, including trophic factor withdrawal, and exposure to oxidative and metabolic insults. Par 4, which can be induced at the translational level, acts at an early stage of the apoptotic cascade prior to caspase activation and mitochondrial dysfunction. The mechanism whereby Par-4 promotes apoptosis may involve inhibition of the antiapoptotic transcription factor NF-kappaB and suppression of Bcl-2 expression and/or function. Studies of postmortem tissues from patients and animal models of neurodegenerative disorders, including Alzheimer's, Parkinson's, and Huntington's diseases, amyotrophic lateral sclerosis (ALS), and HIV encephalitis, have documented increased levels of Par-4 in vulnerable neurons. Manipulations that block Par-4 expression or function prevent neuronal cell death in models of each disorder, suggesting a critical role for Par-4 in the neurodegenerative process. Interestingly, Par-4 levels rapidly increase in synaptic terminals following various insults, and such local increases in Par-4 levels appear to play important roles in synaptic dysfunction and degeneration. A better understanding of the molecular and cellular biology of Par-4 will help clarify mechanisms of neuronal apoptosis, and may lead to the development of novel preventative and therapeutic strategies for neurodegenerative disorders. PMID- 10691288 TI - Inherited and experimentally induced changes in gating kinetics of muscle nicotinic acetylcholine receptor. AB - Ligand-gated ion channels (LGIC) allow rapid responses in the nervous system. The nicotonic acetylcholine receptor (AChR) has been the model for structure-function relationship studies on this superfamily. The AChR undergoes the following functional events: 1. Binding of the neurotransmitter. 2. Opening of the ion channel. 3. Conduction of ions across the pore. 4. Desensitization. The equilibrium among these processes can be perturbed by alteration in the primary structure of the AChR or by the presence of pharmacological agents. Changes in the primary sequence leading to modifications in gating kinetics may occur in association with physiological or pathological processes. Such changes can also be genetically engineered to gain insights into structure-function relationships. PMID- 10691290 TI - The regulation of hippocampal nicotinic acetylcholine receptors (nAChRs) after a protracted treatment with selective or nonselective nAChR agonists. AB - In rats, 1 mg/kg twice daily for 10 d of nicotine, a nonselective agonist of nicotinic acetylcholine receptors (nAChRs), fails to change alpha4 and beta2 nAChR subunit mRNA but significantly decreased alpha7 nAChR subunit mRNA and protein expression, which is associated with a 35-40% decrease in the number of 125I-alpha-Bgtx binding sites in hippocampus. In addition, this schedule of nicotine treatment produced a 40% increase in the number of high- (K(D) 1 nM), but decreased by 25% the number of low-affinity (K(D) 30 nM) binding sites for 3H epibatidine in hippocampus. In contrast, repeated treatment with lobeline (2.7 mg/kg twice daily for 10 d), which selectively binds to high-affinity binding nAChRs, fails to change the expression of high- or low-affinity nAChRs. These data suggest that a simultaneous upregulation of high-affinity nAChRs and downregulation of low-affinity nAChRs is elicited by ligands that can bind to both low- and high-affinity nAChRs, but not by selective agonists of high affinity nAChRs. One might infer that in hippocampus, high- and low-affinity nAChRs may be located in the same cells. When these two receptor types are stimulated simultaneously by nonselective ligands for high- and low-affinity nAChRs, they interact, bringing about an increase in binding site density of the high-affinity nAChRs. PMID- 10691291 TI - Expression of aspartoacylase activity in cultured rat macroglial cells is limited to oligodendrocytes. AB - N-acetyl-L-aspartate (NAA) is an important osmolyte in the vertebrate brain and eye, and its cyclical metabolism is accomplished in two separate compartments. In the brain, NAA is synthesized primarily in neurons, and after its regulated release, NAA is hydrolyzed by aspartoacylase, which is present in a glial associated compartment. However, the precise nature of this hydrolytic compartment has remained obscure. It has been proposed that one role of aspartoacylase in the central nervous system (CNS) is as part of a molecular water pump (MWP) that uses the NAA intercompartmental cycle to remove nerve cell metabolic water against a water gradient and that oligodendrocytes comprise the second compartment in this metabolic sequence. The absence of aspartoacylase activity in Canavan disease (CD), a rare early onset genetic spongiform leukodystrophy, is associated with CNS edema, intramyelinic swelling and a progressive loss of oligdendrocytes. In order to evaluate the MWP hypothesis and its possible relationship to the etiology of CD further, both oligodendrocytes and astrocytes obtained from neonatal rat brain were grown in culture and tested for the presence of aspartoacylase activity. The results of this study show for the first time that aspartoacylase activity is expressed only in oligodendrocytes. The meaning of this observation in understanding the function of the NAA metabolic cycle is discussed. PMID- 10691292 TI - Anandamides inhibit binding to the muscarinic acetylcholine receptor. AB - Loss of memory and cholinergic transmission are associated with both Alzheimer's disease (AD) and marijuana use. The human brain muscarinic acetylcholine receptor (mAChR), which is involved in memory function and is inhibited by arachidonic acid, is also inhibited by anandamides. Two agonists of the cannabinoid receptor derived from arachidonic acid, anandamide (AEA) and R-methanandamide, inhibit ligand binding to the mAChR. Binding of the mAChR antagonist [3H]quinuclidinyl benzilate ([3H]QNB) is inhibited up to 89% by AEA (half-maximal inhibition at 50 microM). Binding of the more polar antagonist [N-methyl-3H]scopolamine ([3H]NMS) is inhibited by AEA up to 76% (half-maximal inhibition at 44 microM). R methanandamide inhibits more than 90% of both [3H]QNB binding (I50 = 34 microM) and [3H]NMS binding (I50 = 15 microM) to the mAChR. Both AEA and R-methanandamide stimulate mAChR binding of the agonist [3H]oxotremorine-M at low concentrations (25-75 microM), but significantly inhibit agonist binding at higher concentrations (I50 = 150 microM). The cannabinoid antagonist SR141716A did not alter AEA or R-methanandamide inhibition of [3H]NMS binding to the mAChR, even at concentrations as high as 1 microM. Further, the cannabinoid agonist WIN 55212-2 does not alter antagonist binding to the mAChR. This demonstrates that mAChR inhibition by the anandamides is not mediated by the cannabinoid receptor. Since AEA and R-methanandamide are structurally similar to arachidonic acid, they may interact with the mAChR in a similar manner to inhibit receptor function. PMID- 10691293 TI - Reactive oxidant species in piriform cortex extracellular fluid during seizures induced by systemic kainic acid in rats. AB - Kainic acid (KA) administered systemically to rats produces seizures and brain damage. We measured an increase in reactive oxidant species (ROS) during KA induced seizures in the extracellular fluid (ECF) of the piriform cortex, a brain region known to be subsequently damaged. Intracerebral microdialysis samples were collected and assayed for isoluminol-dependent chemiluminescence before and after injection of KA (16 mg/kg, i.p.). Hydrogen peroxide (H2O2) concentrations were calculated from catalase-sensitive chemiluminescence, the difference between total and catalase-resistant chemiluminescence. During generalized tonic-clonic seizures, both total and catalase-resistant chemiluminescence increased significantly in samples from brain ECF. Catalase-resistant chemiluminescence, most likely produced by ascorbic acid, increased for a full hour during sustained seizure activity. H2O2 concentrations showed a trend towards elevation during seizures. Increased ROS suggest that oxidative stress occurs in brain ECF during sustained seizure activity. PMID- 10691294 TI - Region-specific changes in prodynorphin mRNA and ir-dynorphin A levels after kindled seizures. AB - The opioid peptide dynorphin is thought to be implicated in specific types of seizures. In particular, complex partial seizures have been shown to cause release of dynorphin, activation of prodynorphin gene expression, and new peptide synthesis in the hippocampus. In this study, the kinetics of the seizure-induced changes in prodynorphin mRNA and ir-dynorphin A levels in the hippocampus have been compared with those induced in the temporal and frontal cortex, i.e., in other regions involved in the pathophysiology of complex partial seizures. Experiments have been run using kindling, one of the most valuable models of partial epilepsy. In the hippocampus (1) prodynorphin mRNA levels transiently increase (threefold) 1 h after kindled seizures, and return to baseline by 2 h, and (2) dynorphin A levels are slightly decreased at 1 h, but increase (twofold) at 2 h and return to baseline by 6 h. In the temporal and in the frontal cortex, a late (beginning at 2 h) and prolonged (up to 24 h) decrease in both prodynorphin mRNA and ir-dynorphin A levels have been observed. These data suggest that differential changes in dynorphin metabolism occur in different brain areas after seizures. The mechanisms and functional implications of this observation remain to be investigated. PMID- 10691295 TI - Increased vulnerability of neuronal cell lines to sodium nitroprusside-mediated toxicity is caused by the decreased level of nitric oxide metabolites. AB - Nitric oxide (NO) is an unstable radical produced during the oxidative deamination catalyzed by NO synthase (NOS) that converts L-arginine to L citrulline. NO is also generated nonenzymatically from a group of compounds, called NO donors, such as sodium nitroprusside (SNP). NO directly or through its metabolites has been implicated in several disorders, including Alzheimer's disease (AD). Since NO is a highly labile unstable free gas, we measured the stable end products, nitrite and nitrate (NOx). Here, we investigated the effect of SNP-mediated NO release in different cell types and its effect on the beta amyloid precursor protein (betaAPP). When different cell types were induced with SNP, a significant level of NOx was detected in a time and dose-dependent manner over the spontaneous release of NOx by SNP. The astrocytes, glial, and epithelial cell lines released significantly higher level of NOx as compared to neuronal cells following the exposure of SNP. The latter group of cells was more sensitive to NO-mediated cytotoxicity, as demonstrated by the lactate dehydrogenase assay. The SNP-mediated toxicity is known to be caused by the accumulation of cyanide ions and we report that the ability of cells to protect against it depends on the levels of nitric oxide metabolites. Cell lines, such as astrocytic and epithelial, that produce more NOx are better protected against the SNP-induced toxicity than the less NOx-protecting neuronal cell lines. The possibility of differential susceptibility of neurons and astrocytes resulting from the different content of reduced glutathione is also discussed. The release of NOx was prevented by cotreatment with a NO scavenger and superoxide dismutase but not by a NOS inhibitor. The activity of NOS was decreased when cytosolic extracts were incubated with SNP. In the conditioned medium of SNP-induced cells, the level of soluble betaAPP (sAPP) was decreased, and this decrease was more apparent in neuronal than astrocytic cell lines. Taken together, these results suggest that the SNP-derived NO release is independent of the NOS pathway, that various cell types metabolize SNP differently, and that neuronal cell lines are more vulnerable with SNP treatment with lowered sAPP secretion. Since the neuronal cell lines lack a nitric-oxide-generated protective mechanism, we speculate that these cells may be the first targets of neurodegeneration by several toxic agents, including the cyanides and peroxynitrites. PMID- 10691296 TI - Histamine release from mast cells of EAE rats by Gi protein-dependent and IgE dependent pathways. AB - We investigated both Gi protein-dependent and IgE-dependent pathways that control release of histamine by PMCs derived from EAE or Complete Freund's Adjuvant (CFA) immunized rats. The number and histamine content of MCs per rat were the same between normal and EAE rats. Activation of Gi pathway by substance P (SP), DSA, 48/80, and mastoparan resulted in a dose-dependent increase in release of histamine by PMCs in normal, EAE-, and CFA-immunized rats. In EAE and CFA rats, however, the induction was decreased by 10-20% compared to normal rats. The histamine release induced by MP was decreased at a concentration of 3 microM, but not at 10 microM in severe active EAE rats. Activation of the IgE pathway by MAM and concanavalin A (Con A) in the presence of phosphatidylserine led to dose dependent histamine release in normal rats, and a 10-25% lower level of induction was observed in EAE rats. In CFA rats, the induction of histamine release was equivalent to normal rats. There was an increase in intracellular calcium stores following activation of both pathways in normal rats, whereas depletion of calcium stores by ryanodine reduced the level of induction by 48/80 and MP by 9 11% in normal rats. In EAE rats, 48/80, Con A, and MAM induced a smaller increase, but SP and MP induced larger or similar increases in calcium stores compared to normal rats. It was unlikely that the calcium stores of the PMCs from EAE rats were depleted, because MP stimulated calcium movement subsequent to the release of histamine. These results suggested that the Gi pathway may not be correlated to clinical manifestation of EAE, but cold be involved in the inflammatory process, and that the IgE pathway is better associated with clinical symptoms of EAE and may be more directly related to disease outcome. PMID- 10691297 TI - The growth-associated protein GAP-43 is increased in the hippocampus and in the gyrus cinguli in schizophrenia. AB - Schizophrenia is a common and severe psychiatric disorder of unknown etiology. Numerous neuropathological studies have found subtle structural changes in limbic structures, especially medial temporal lobe structures and the gyrus cinguli. To test the hypothesis that synaptic disturbances are involved in the pathogenesis of schizophrenia, we studied the growth-associated protein 43 (GAP-43), a protein localized to presynaptic terminals, suggested to be involved in establishment and remodeling of synaptic connections, in postmortem brain tissue, using quantitative Western blotting immunohistochemistry. The material consisted of brain tissue from 17 schizophrenics (80 +/- 11 yr), diagnosed according to the DSM-III-R criteria, and 20 age-matched controls (75 +/- 13 yr). Quantitative analyses showed increased GAP-43 protein levels in schizophrenic compared to control brains, both in the hippocampus (2.43 +/- 0.78 vs 1.00 +/- 0.29; p < 0.0001) and in the gyrus cinguli (1.52 +/- 0.21 vs 1.00 +/- 0.35; p < 0.0001). Also by immunohistochemistry, increased GAP-43 staining was found in schizophrenic compared with control brains, throughout all layers of the gyrus cinguli and the hippocampus. Anomalous synaptic sprouting and reorganization, with resultant "miswiring," as well as a defect in synaptic pruning have been hypothesized to be pathogenetic factors in schizophrenia. We suggest that a decreased synaptic density, whether caused by disturbed development or damage/degeneration, may elicit a reactive synaptogenesis (reflected by an increase in GAP-43), which may be functional or anomalous. Synaptic pathology in the limbic system may be of importance in the development of psychotic symptoms in schizophrenia. PMID- 10691299 TI - Increased expression of prion protein is associated with changes in dopamine metabolism and MAO activity in PC12 cells. AB - Prion diseases of humans and animals occur following infection with infectious agents containing PrP(Sc) or in situations in which there is a mutation of the prion protein (PrP) gene. The cellular prion protein (PrP(C)) is a sialoglycoprotein that is expressed predominantly in neurons. PrP(C) is converted into a pathogenic form of PrP (PrP(Sc)), which is distinguishable from PrP(C) by its relative resistance to protease digestion. A number of postulates have been advanced for the function of normal PrP (PrP(C)), but this issue has not been resolved. To investigate the function(s) of PrP(C), we established clonal PC12 cell lines, which have elevated PrP(C) expression. The results show that there were alterations in dopamine metabolism and in monoamine oxidase (MAO) activity in transfected PC12 cells that overexpress PrP(C). There was an increase in concentration of DOPAC, a metabolite of dopamine, and in MAO activity in cells overexpressing PrP(C). MAO is involved in oxidative degradation of dopamine (DA). Our data suggest that PrP(C) plays a role in DA metabolism by regulating MAO activity. PMID- 10691298 TI - Mice transgenic for a human amyloid precursor protein promoter-lacZ reporter construct. AB - Transgenic mouse lines were generated that expressed a 2-kb amyloid precursor protein (APP) promoter/beta-galactosidase reporter gene construction. In brain, hippocampal pyramidal neurons, neurons in the deeper layers of cerebral cortex, and neurons in several thalamic nuclei were heavily labeled by beta-galactosidase histochemistry. In general, molecular layers and white matter regions did not express the reporter gene. When compared with in situ hybridization for endogenous murine APP RNA, the striatum and outer layers of cerebral cortex had little reporter expression. Thus, the match between reporter expression and endogenous APP expression in brain was not perfect. A similar mismatch between the relative expression of the reporter gene and endogenous APP RNA distribution was found in homogenates from several organs. Although prior work in transgenic mice found similar mismatches in reporter gene distribution, none had tested the APP promoter construct in response to neuronal injury. Kainic acid injections successfully increased murine APP expression in the transgenic mice, but had no effect on the reporter gene expression. Based on these data and those collected by others, we conclude that the 2-kb region upstream of the APP transcription initiation site contains some elements responsible for the tissue-specific expression of this gene, but does not contain all the cis-acting elements sufficient for either the differential tissue distribution of this gene or the regulation of this gene subsequent to neural damage. PMID- 10691300 TI - Chronic lead intoxication affects the myelin membrane status in the central nervous system of adult rats. AB - The aim of the experiments presented here was to discern whether prolonged consumption of leaden water, which imitates an environmental exposure, affects the structure of myelin in the central nervous system of adult rats and whether the observed morphological destruction is reflected in biophysical and/or biochemical changes. The results indicated that during chronic lead (Pb) intoxication, the Pb level of the myelin fraction increases significantly. Electron microscopy studies show that myelin in control experiments is built up of ordered layers, whereas in a Pb-intoxicated sample, this order is destroyed in large areas of all preparations. Morphological disturbances in Pb-intoxicated in vivo myelin were reflected by changes in myelin membrane fluidity measured by spectrofluorometry and electron paramagnetic resonance (EPR). Prolonged Pb toxicity also caused significant changes in the morphological structure of oligodendrocytes, an increase of phosphatidylethanolamine, and decrease of protein SH group levels. Simultaneously, we found that the protein and total phospholipid content and levels of phosphatidylinositol, sphingomyelin, phosphatidyloserine, cholesterol, and the pattern of total myelin protein obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in Pb-intoxicated myelin did not change compared to control values. Also, Pb intoxication did not induce peroxidation by itself and did not accelerate peroxidation produced by iron (Fe) in brain myelin. PMID- 10691301 TI - Activity-dependent interaction of the intracellular domain of rat trkA with intermediate filament proteins, the beta-6 proteasomal subunit, Ras-GRF1, and the p162 subunit of eIF3. AB - Many responses to nerve growth factor (NGF) are regulated through the receptor tyrosine kinase trkA. To understand more fully the functions of trkA in NGF responsive cells, we have expressed the intracellular domain of rat trkA as a fusion protein with the yeast gal4 transcription factor, and used the fusion protein to probe rat and mouse cDNA libraries by the yeast two-hybrid system. We have identified a direct interaction between the intracellular domain of trkA and two members of the intermediate filament (IF) family of proteins, the guanine nucleotide exchange protein Ras-GRF1, the p162 subunit of eIF3, and the beta-6 proteasome subunit. The interactions are dependent on an active trkA kinase, and RasGRF1, the beta-6 proteasomal subunit, and peripherin are directly phosphorylated by trkA. The interaction with trkA is not affected by mutations at either Tyr499 or Tyr794, the two major phosphotyrosine residues essential to the activation and receptor binding of Shc, FRS-2/SNT, and phospholipase Cgamma-1, and it is highly specific in vitro for trkA, with little or no binding observed with trkB and/or trkC. The results show that trkA may play a regulatory role in a variety of cellular functions in addition to neuritogenesis, including regulated protein degradation and transcriptional activation. PMID- 10691302 TI - Aberrant stress response associated with severe hypoglycemia in a transgenic mouse model of Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) exhibit alterations in glucose metabolism and dysregulation of the stress-responsive hypothalamic-pituitary-adrenal (HPA) neuroendocrine system. The mechanisms responsible for these alterations and their possible contributions to the neurodegenerative process in AD are unknown. We now report that transgenic mice expressing a mutant form of human amyloid precursor protein (APP) that causes inherited early-onset AD exhibit increased sensitivity to physiological stressors, which is associated with aberrancies in HPA function and regulation of blood glucose levels. Specifically, APP mutant mice exhibit severe hypoglycemia and death following food restriction, and sustained elevations of plasma glucocorticoid levels and hypoglycemia following restraint stress. The alterations in HPA function and glucose regulation were evident in relatively young mice prior to overt deposition of amyloid beta-peptide (A beta). However, diffuse accumulations of A beta were present in the hypothalamus of older mice, suggesting a role for soluble forms of A beta in dysregulation of HPA function. Our data demonstrate disturbances in neuroendocrine function in APP mutant mice similar to those seen in AD patients. These impairments in stress response, glucocorticoid signaling, and regulation of blood glucose should be considered in interpretations of data from past and future studies of APP mutant mice. PMID- 10691303 TI - Immunolocalization of tenascin-C in human type II fiber atrophy. AB - Tenascin-C is a multifunctional extracellular matrix glycoprotein with stimulatory and anti-adhesive or inhibitory properties for axon growth. Its location and discontinuous expression are restricted in innervated muscle tissues. Tenascin-C accumulated interstitially among human denervated muscle fibers and close to normal-sized fibers. To expand our knowledge of the expression of tenascin-C in human neuromuscular disorders, we investigated immunohistologically 20 human muscle specimens with type II myofiber atrophy of children and adults. Tenascin-C immunoreactivity in adult type II atrophy was frequent, and accumulation in children was sparse and weak. In both groups, tenascin-C immunoreactivity was found: 1. Interstitially around normal-sized type II muscle fibers. 2. Around atrophic type II muscle fibers. 3. Around small caliber myofibers with centrally located nuclei. These results indicate that tenascin-C immunoreactivity: (1) is detectable around early denervated and reinnervated muscle fibers and, therefore, (2) may reflect in part the molecularly ongoing process of denervation and reinnervation in human type II fiber atrophy. PMID- 10691304 TI - Expression of vascular endothelial growth factor (VEGF) and platelet-derived growth factor receptor-beta (PDGFR-beta) in human gliomas. AB - The growth of solid tumors is highly dependent on vascular proliferation. Vascular endothelial growth factor (VEGF), the main mediator of angiogenesis, and platelet-derived growth factor receptor-beta (PDGFR-beta), receptor for the potent mitogen PDGF, are two indicators of the angiogenic potential of human gliomas. We studied a series of 57 surgical biopsies of astrocytic neoplasms by immunohistochemistry to elucidate the relationship between tumor proliferation, quantified as Ki67-LI, and the expression of these two proteins. Ki67-LI increases throughout histological malignancy, although staining in endothelial cells has rarely been recorded. Elevated amounts of VEGF-positive tumor cells (VEGF-LI) were found in anaplastic astrocytomas and glioblastomas, mainly around areas of necrosis, cysts, or edema. Endothelium of blood vessels was consistently stained. PDGFR-beta positivity was found in glomeruloid formations and in tumor cells, excluding pilocytic astrocytomas. Multinucleated giant cells and perivascular tumor cells were positive in glioblastomas. In addition, peritumoral microglia-like cells were also stained in some cases. Statistical correlation was only found between PDGFR-beta and Ki67 LIs. In conclusion, VEGF as permeability factor is involved in the development of secondary neoplastic changes, whereas PDGFR-beta is directly correlated to proliferation indexes. Strong expression of VEGF and PDGFR-beta found in endothelium and tumor cells would seem to support a combined role in tumoral neoangiogenesis. PMID- 10691305 TI - Fibroblast growth factor receptor 4 (FGFR4) is expressed in adult rat and human retinal photoreceptors and neurons. AB - The fibroblast growth factor (FGF) family, with its prototype members acidic FGF (FGF-1) and basic FGF (FGF-2), binds to four related receptor tyrosine kinases, termed FGFR1, R2, R3, and R4, expressed on most types of cells in tissue culture. In many respects, the FGFR appear similar to other growth factor receptors; thus, dimerization of receptor monomers on ligand binding is likely to be a requisite for activation of the kinase domains, leading to receptor trans-phosphorylation. Within the central nervous system (CNS), including retina, FGFR1 and R2 have been widely described as the predominant forms. FGFR4 is reported to be strongly expressed only during early stages of development, and apart from one small region (the lateral habenular nucleus) is not detectable in adult CNS. Screening of different neural and nonneural tissues by reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that whereas FGFR1 and R2 were strongly expressed in adult cortex, cerebellum, retina, and kidney, robust FGFR4 expression was only seen in retina and kidney. FGFR4 mRNA was present within fractions of the outer and inner nuclear layers isolated from adult rat retinas, and could also be detected in pure photoreceptor cultures prepared from young rat retinas. On the contrary, FGFR4 mRNA could not be detected in primary cultures of retinal Muller glia or pigment epithelium, indicating specific enrichment in retinal neurons. In situ hybridization studies of adult rat retina showed FGFR4 expression in all retinal cellular layers, especially prominent in the outer nuclear layer. FGFR4 protein was detected by immunoblotting of homogenates of rat retina, with specific antibody binding to bands at 115, 47, and 30 kDa. FGFR4 mRNA and protein were also reliably detected in postmortem adult human retina. The potential roles of these signal transduction molecules in FGF-induced biological responses in the retina are discussed. PMID- 10691307 TI - Substitutions of tyrosine 601 in the human thyrotropin receptor result in increase or loss of basal activation of the cyclic adenosine monophosphate pathway and disrupt coupling to Gq/11. AB - Constitutively activating mutations of the thyrotropin (TSH) receptor have been identified as a molecular cause of toxic adenomas, nonautoimmune familial hyperthyroidism, and sporadic congenital hyperthyroidism. By analyzing genomic DNA from a toxic adenoma, we detected a novel somatic mutation in codon 601, tyrosine to asparagine (Y601N), a residue located in the carboxyterminal part of the fifth transmembrane helix. This codon is also notable for the presence of a polymorphic variant, Y601H. These two naturally occurring substitutions (Y601N and Y601H) were analyzed together with an artificial mutation, Y601F, to study the role of this residue for receptor function further. Transient transfection assays revealed that the Y601N mutation results in constitutive activation of the cyclic adenosine monophosphate (cAMP) pathway, but that it is unable to couple to Gq/11. Y601H and Y601F do not display basal activity while retaining responsiveness to TSH, but also lose the ability to induce inositol phosphate accumulation in response to TSH. These studies define Y601N as a mutation that selectively activates the cAMP pathway, and they confirm that Y601H is not a silent polymorphism. In conclusion, residue Y601 has an important role for the characteristic constitutive basal activity of the TSH receptor and coupling to Gq/11. PMID- 10691306 TI - Activity-dependent neurotrophic factor-14 requires protein kinase C and mitogen associated protein kinase kinase activation to protect the developing mouse brain against excitotoxicity. AB - Activity-dependent neurotrophic factor (ADNF) is a newly identified compound that prevents in vitro neuronal death when present in fentomolar concentrations. ADNF 14, a 14 amino acid peptide derived from ADNF, has the same effects on growth as the parent molecule. However, the transduction pathways and target cells for these highly potent trophic factors are still unknown. We previously described a mouse model of excitotoxic lesions of the developing neocortex mimicking several hypoxic or hypoxic-like brain lesions observed in human fetuses and neonates. In this model, cotreatment with the excitotoxin ibotenate and ADNF-14 prevented both neuronal death in pups injected on the day of birth and white matter cystic lesions in pups treated 5 d after birth. In the present study, coadministration of ibotenate, ADNF-14, and selective transduction pathway inhibitors showed that activation of protein kinase C (PKC) and mitogen-associated protein kinase kinase was critical for neuroprotection. Immunocytochemistry revealed that ADNF-14 activated PKC and mitogen-associated protein kinase in cortical neurons on the day of birth and in white matter astrocytes on the fifth postnatal day. Taken in concert, these data identify PKC and mitogen-associated protein kinase pathways as critical to ADNF-14-induced neuroprotection of the developing brain against excitotoxic damage. PMID- 10691308 TI - Quantitative analysis of DNA binding affinity and dimerization properties of wild type and mutant thyroid hormone receptor beta1. AB - Thyroid hormone (triiodothyronine [T3]) actions are mediated through binding of thyroid hormone receptors (TRs) to specific DNA sequences (thyroid hormone response elements [TREs]) as monomers, homodimers, and heterodimers with thyroid hormone receptor auxiliary proteins (TRAPs). We quantitatively characterized dimerization of wild-type (WT) and mutant TRbetas by coimmunoprecipitation, and binding to DNA by electrophoretic gel mobility shift assays (EMSA). Binding affinities of TR retinoid X receptor-alpha (RXRalpha) heterodimers to DNA were determined by competing with excess nonradiolabeled TREs in EMSA. TRs in vitro synthesized in reticulocyte lysates (RL), and human RXRalpha expressed in a Sf9 cell-baculovirus system (BAC), were coincubated with 32P-labeled rat malic enzyme (ME), palindromic (PAL), or chicken lysozyme F2 (F2) TREs. The mutant TRbetas tested were R316H and G345R, which have nondetectable T3 binding and have previously been reported to show weak and potent dominant negative effect, respectively. Scatchard analysis showed no significant differences in Kas between WT and mutant TR-RXRalpha heterodimers binding to DNA. We measured affinity of heterodimerization between TRs and RXRalpha in solution in the absence of DNA, and by coimmunoprecipitation using anti-TRbeta1WT specific antibodies. 35S labeled RL-RXRalpha was incubated with BAC-WT or TRbeta or R316H in the absence or presence of increasing amounts of nonlabeled BAC-RXRalpha. Displacement curves were obtained by counting radioactivity of precipitated 35S-RXRalpha, that was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. Kds of WT and TRbeta R316H heterodimerizing with RXRalpha were approximately the same. Binding affinity of TR homodimers for F2-TRE was studied because this TRE binds homodimers strongly. Scatchard analysis clearly showed that DNA binding affinity of BAC-WT homodimers did not differ with or without 100 nM T3, but maximal binding capacity (MBC) was reduced three-fold to fourfold in the presence of 100 nM T3. In contrast, BACTRbeta-R316H homodimers showed a fivefold reduction in DNA binding affinity for F2, both in the presence and absence of T3, and approximately the same MBC as WT in the absence of T3. Mutant RL-G345R homodimers showed approximately the same Ka as RL-WT homodimers for binding to F2 and the same MBC in the presence and absence of T3. These results indicate that (1) T3 reduced TRbeta homodimerization in solution but does not effect DNA binding of formed homodimers; (2) T3 does not influence DNA binding affinity of TR/RxR heterodimers; and (3) TRbeta mutant R316H homodimers have reduced DNA binding affinity but homodimerization and heterodimerization in solution does not differ from WT TRbeta. PMID- 10691309 TI - N-ras mutation in poorly differentiated thyroid carcinomas: correlation with bone metastases and inverse correlation to thyroglobulin expression. AB - Codon 61 of the N-ras oncogene was screened for mutations in 99 surgically resected thyroid carcinomas by a polymerase chain resection (PCR)-based method (PCR-primer introduced restriction with enrichment of mutant alleles [PCR PIREMA]). A point mutation of the N-ras oncogene at the codon 61 was detected in 16 of 99 (16.2%) thyroid carcinomas examined by this method. No RAS alteration was detected in the group of 11 medullary thyroid carcinomas, while 3 of 31 (10.0%) papillary carcinomas, 2 of 5 (40%) follicular carcinomas, 8 of 44 (18.2%) poorly differentiated carcinomas, and 3 of 5 (60%) undifferentiated carcinomas showed an activation of N-RAS proto-oncogene. Interestingly, two primary follicular tumors and their corresponding bone metastases, showed N-ras mutations. In the same cases we evaluated the expression of thyroglobulin by immunohistochemical analysis. Although the majority of well-differentiated carcinomas expressed a high level of thyroglobulin, the expression of the same antigen was absent or only occasional weakly positive in 33 of 44 poorly differentiated carcinomas. Interestingly, N-ras mutation was restricted to the group of tumours with low or absent thyroglobulin expression, suggesting that this genetic change is prevalent in less differentiated tumors. PMID- 10691310 TI - Molecular alterations involving p53 codons 167 and 183 in papillary thyroid carcinomas from chernobyl-contaminated regions of belarus. AB - After the Chernobyl accident in 1986, there was a significant increase in the incidence of papillary thyroid carcinoma in fallout-exposed children from Belarus. We studied the p53 gene from 24 papillary thyroid carcinoma cases presenting in 1996. All subjects lived in contaminated regions of Belarus at the time of the accident and were under age 20 when exposed to fallout. Exons 5 through 9 of p53 were amplified from genomic tumor DNA using the polymerase chain reaction (PCR). The PCR products were analyzed by direct DNA sequencing using an automated sequencer. Five cases each exhibited two molecular alterations within exon 5. Alterations were confirmed by sequencing in both directions. One alteration, involving codon 167 (CAG-->CAT) in all five cases, resulted in the substitution of HIS for GLN. The second alteration, involving codon 183 (TCA- >TGA) in all five cases, resulted in a premature termination codon. Leukocyte DNA from each of the positive cases was analyzed and found to contain only wild-type p53 sequence. These results suggest that mutations involving codons 167 and 183 in the p53 locus are important in the pathogenesis of a subset (21%) of radiation induced papillary thyroid carcinomas from Belarus. PMID- 10691311 TI - Accuracy of free thyroxine measurements across natural ranges of thyroxine binding to serum proteins. AB - Systemic inaccuracies, proportional to the concentrations of serum proteins and the thyroxine (T4) they carry, have been reported in direct free T4 immunoassays. However, analytical recoveries of free T4 have not been carefully examined in most current methods, and they have not previously been examined across the pathophysiological range of serum T4 binding. In the present study we investigated ranges of serum T4 binding using free and total T4 measurements from 1359 individuals. Carefully characterized, gravimetrically calibrated, serum based free T4 test solutions were then prepared with a constant normal free T4 concentration (12 ng/L) and varied serum T4 binding (approximately 300:1 to 24,000:1, ng protein bound T4: ng free T4). These standardized test solutions were analyzed using five T4 analog based free T4 methods. Analytical recoveries were calculated as ratios of actual free T4 measurements to the target value, and expressed as a percent of the target. Analytical recoveries were directly proportional to the extent of serum T4 binding and ranged 2% to 155%, 25% to 131%, 53% to 106%, 37% to 93%, and 37% to 73%, lowest to highest, in different methods. These systemic inaccuracies will confound interpretations of free T4 test results in clinical conditions with altered T4 binding. Future investigations into free T4 status must examine the analytical recovery of the free T4 method(s) used, as they relate to the extent of serum T4 binding in the clinical condition(s) studied. PMID- 10691312 TI - Functions of thyroid hormone receptors in mice. AB - Thyroid hormone receptors (TRs) play a central role in mediating the actions of thyroid hormone in development and homeostasis in vertebrate species. The TRs are nuclear receptors that act as ligand-regulated transcription factors. There are two TR genes (TRalpha and TRbeta), each capable of generating different variant products, suggesting a potentially complex array of TR pathways. Targeted mutagenesis in the mouse has indicated that there are specific individual functions for the TR genes in vivo. The deletion of combinations of TRalpha and TRbeta variants has revealed that additional functions are convergently regulated by both TR genes and indicates that control of an extended range of functions is facilitated by a network of specific and common TR pathways. The TR-deficient mouse models have allowed investigation of the TR pathways underlying many functions of thyroid hormone and provide a unique perspective on receptor mediated mechanisms of biological control. PMID- 10691313 TI - Preparation and culture of a serum-free human thyroid follicle system and its application for measuring thyroid hormone secretion, iodide uptake and organification, cyclic adenosine monophosphate formation, gene expression, and cell growth. AB - We describe a system of human thyroid follicles cultured in collagen suspended in serum-free medium. The method allows measurement of thyroid hormone secretion, iodide uptake and organification, cyclic adenosine monophosphate (cAMP) formation, gene expression, as well as cell growth. The system is superior to follicles freely suspended in cultured medium and also, as demonstrated by parallel experiments, to monolayer culture systems. A detailed description of the optimal conditions of the method is provided that, over a period of 8 years, has proven to be a powerful tool for measuring thyroid cell function, gene expression and cell proliferation. PMID- 10691314 TI - Potential repercussions for the progeny of maternal hypothyroxinemia during pregnancy. PMID- 10691315 TI - Fine-needle aspiration of thyroid nodules in radiation-exposed patients. AB - External radiation used to treat benign conditions in the head and neck area results in an increased risk of thyroid cancer in exposed individuals. Fine needle aspiration (FNA) biopsy is the standard procedure used to evaluate suspicious thyroid nodules. Its accuracy has been extensively studied, but little is known about FNA in irradiated patients. We analyzed the FNA experience of 136 irradiated subjects. Fifty-two had surgery enabling a comparison of the histologic diagnosis with the FNA results. In these 52 patients with a total of 53 FNAs, 20 were reported as benign, 14 as follicular neoplasms, 6 as papillary cancer, and 13 as inadequate samples. Seven malignant nodules were aspirated; 4 were reported as papillary cancer, 1 was reported as benign and 2 had inadequate specimens. An additional 11 patients had thyroid cancer in foci that were not subjected to FNA. For the nodules that were aspirated, and considering an FNA report of follicular neoplasm as a false-positive when a follicular adenoma or a colloid nodule was found at surgery, the calculated sensitivity was 80%, specificity 54%, positive predictive value 20%, and negative predictive value 95%. Of the 14 follicular neoplasm FNA diagnoses, 10 were colloid nodules (71%), and 4 only were follicular adenomas. We conclude that the sensitivity of FNA in irradiated patients is similar to what is reported for the general population. However, smaller malignant nodules are common and are not diagnosed by the FNA. Also, the FNA diagnosis of follicular neoplasm is often inaccurate and inadequate aspirations are frequent in this patient group. PMID- 10691316 TI - Postpartum thyroiditis: epidemiology and clinical evolution in a nonselected population. AB - Postpartum thyroiditis (PPT) presents in approximately 5% of women. Its incidence, clinical characteristics, and evolution were studied in a nonselected population of Mediterranean women. Six hundred five healthy women, recruited between the 36th week of pregnancy and the 4th postpartum day, underwent initial clinical and biological evaluation and postpartum at 1 (n = 605), 3 (n = 552), 6 (n = 574), 9 (n = 431), and 12 (n = 444) months. PPT was diagnosed in women with transient hyperthyroidism between 1 and 3 months postpartum and/or hypothyroidism between 3 and 6 months postpartum. Permanent hypothyroidism was considered if it was overt and persisted one year after diagnosis. The incidence rate of PPT was 7.8%. Eighty-two percent of PPT patients had hormone abnormalities at the 6th month postpartum, 8.8% showed depression and 51% goiter. PPT was manifest as hyperthyroidism plus hypothyroidism in 35.5% of patients, because only transient hyperthyroidism in 22.2% and as hypothyroidism alone in 42.3%. Five patients with hypothyroidism during PPT (0.82% of the initial population, 11.1% of PPT patients, and 15.6% of hypothyroidism PPT patients) presented permanent hypothyroidism after a follow-up of 39.8 (4.2) months. PPT was found in 7.8% of general Mediterranean population. We recommend evaluation at the 6th postpartum month to diagnose the majority of PPT women and indefinite follow-up of hypothyroid PPT patients to detect permanent hypothyroidism. PMID- 10691317 TI - The blood spot thyrotropin method is not adequate to screen for hypothyroidism in the elderly living in abundant-iodine intake areas: comparison to sensitive thyrotropin measurements. AB - We investigated whether the blood spot thyrotropin (TSH) method was adequate for screening elderly subjects with abundant iodine intake (median excretion 330 microg/g creatinine) for hypothyroidism. In 97 healthy adults (group A), 210 nursing home residents (group B) and 265 elderly subjects living at home (group C) serum (sensitivity < 0.02 mU/L, cost 1.2 U.S. dollars [USD]) and blood spot TSH (sensitivity < 1.0 mU/L, cost 0.4 USD) were measured, and the sensitivity and specificity of different blood spot TSH cutoff points to detect cases with elevated serum TSH were calculated. Elevated (> 3.5 mU/L) serum TSH levels (group A, 6.2%; group B, 16.2%; group C, 22.3%; B > A, p = 0.025; C > A, p < 0.001) were detected with the required sensitivity of greater than 0.9 only if the cutoff point of the blood spot TSH was set as low as 2.5 mU/L, but this led to a considerable loss of specificity. At cutoff point 2.5 mU/L, the rate of positivity was 39.3% and the cost of blood spot screening/person increased to 0.88 USD, considering that positive cases have to be rechecked by serum TSH to exclude false positivity. Cases with significantly elevated (> 10.0 mU/L) serum TSH (group A, 1.03%; group B, 2.85%; group C, 2.20%) were detected at blood spot cutoff points 10.0-4.0 mU/L with a sensitivity of 1.0 and without considerable loss of specificity. We conclude that while screening for hypothyroidism in the elderly population with abundant iodine intake is justified by the high prevalence of elevated ultrasensitive serum TSH values, the sensitivity of the blood spot method is insufficient to detect the subclinical hypothyroidism accurately and would, therefore, fail to detect most affected subjects. PMID- 10691318 TI - Administration of interferon-gamma in healthy subjects does not modulate thyroid hormone metabolism. AB - Cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interleukin (IL-2), IL-6, and interferon-alpha (IFN-alpha), alter human thyroid hormone metabolism and may be involved in the pathogenesis the euthyroid sick syndrome. Experimental data suggest that interferon-gamma (IFN-gamma) could be another cytokine that might influence thyroid hormone metabolism. To evaluate whether IFN-gamma can be involved in the pathogenesis of the alterations in thyroid hormone metabolism in humans with nonendocrine illness, we measured thyroid hormone concentrations in six healthy volunteers during 24 hours in a placebo controlled trial: once after subcutaneous administration of IFN-gamma (Immukine, [Boehringer Ingelheim GmbH, Ingelheim/Rheim, Germany] 100 microg/m2 subcutaneous) and once after the administration of saline (control). In addition, we measured cytokine concentrations in plasma (TNF-alpha and IL-6). IFN-gamma did not induce effects on any of the measured thyroid hormone and thyroid-stimulating hormone (TSH) plasma concentrations. Moreover, IFN-gamma did not affect TNF-alpha plasma levels. Only a modest but significant elevation of plasma IL-6 levels was detected after administration of IFN-gamma (p < 0.05 vs. control). It is concluded that IFN-gamma administration to healthy humans does not result in short term alterations of thyroid hormone metabolism. These data do therefore not support a role of IFN-gamma in the pathogenesis of the euthyroid sick syndrome in humans as might be deduced from in vitro and in vivo animal studies. PMID- 10691319 TI - Unusual malabsorption of levothyroxine. AB - We report a 50-year-old woman, with overt hypothyroidism undergoing thyrotropin (TSH)-stimulating hormone suppressive levothyroxine (LT4) treatment after subtotal thyroidectomy. At her first visit to our department, the laboratory results revealed a borderline low free thyroxine (FT4) level accompanied by a clearly elevated TSH level. Both parameters did not significantly change during therapy with an oral dose of 500 microg of LT4. Investigations revealed malabsorption of oral administrated LT4. Thyroid serum hormone levels only became normal during parenteral therapy with LT4. PMID- 10691320 TI - Managed care and the practicing thyroidologist: an overview. PMID- 10691321 TI - Unilateral ophthalmopathy in a patient with Hashimoto's thyroiditis. PMID- 10691322 TI - Thyrotoxicosis after interferon-alpha therapy. PMID- 10691323 TI - Use of colony-stimulating factors for the treatment of antithyroid drug-induced agranulocytosis: a retrospective study in twelve patients. PMID- 10691324 TI - Bucharioside and buchariol from Salvia bucharica. AB - A new monoterpene-glycoside (2-exo-beta-D-glucopyranosyl-1,8-cineol) named bucharioside from the methanol-soluble part and a new sesquiterpenoid (4,10-epoxy 6alpha-hydroxyguaiane) named buchariol from the hexane-soluble part of Salvia bucharica were obtained. Their structures were elucidated with the help of NMR spectroscopy including 1D and 2D experiments. PMID- 10691325 TI - [Treatment of gastric phytobezoars with cellulase]. PMID- 10691326 TI - Metabolic interactions between mibefradil and HMG-CoA reductase inhibitors: linking in vitro with in vivo information. PMID- 10691327 TI - Oochroconis gallopava endophthalmitis in fludarabine treated chronic lymphocytic leukaemia. PMID- 10691328 TI - Vancomycin and ceftazidime incompatibility upon intravitreal injection. PMID- 10691329 TI - Acute zonal occult outer retinopathy. PMID- 10691330 TI - Linear naevus sebaceous syndrome, optic disc staphyloma, and non-rhegmatogenous retinal detachment. PMID- 10691331 TI - Can leucocoria be the first manifestation of protrin C deficiency? PMID- 10691332 TI - Proceedings of the Sheffield Meeting of the Society for Research into Hydrocephalus and Spina Bifida. 1999. PMID- 10691333 TI - Photo quiz. Infection due to Exophiala jeanselmei. PMID- 10691334 TI - Role of muscle-derived cells in hematopoietic reconstitution of irradiated mice. PMID- 10691335 TI - Inheritance of chromosomally integrated viral DNA? PMID- 10691336 TI - Detection of vascular endothelial growth factor in AIDS-related primary effusion lymphomas. PMID- 10691337 TI - X-linked thrombocytopenia identified by flow cytometric demonstration of defective Wiskott-Aldrich syndrome protein in lymphocytes. PMID- 10691338 TI - Proceedings of the 5th International Potsdam Symposium on Tick-borne Diseases: Tick-borne Encephalitis and Lyme Borreliosis. Berlin, Germany, February 26-27, 1999. PMID- 10691339 TI - Commentary on Garety and Freeman II: 'Cognitive approaches to delusions--A critical review of theories and evidence'. PMID- 10691341 TI - Leishmania braziliensis promastigotes and amastigotes interact differently with host macrophages. AB - In vitro and in vivo ultrastructral studies reveal that the parasite entrance into the macrophage occurs by phagocytosis. The early stage of phagocytosis exhibited different ultrastructural characteristics in both forms of the parasite. Long and prominent projections from peritoneal exudate macrophages made focal contacts with the promastigote surface. The amastigotes, in turn, laid on cup-shaped extensions of the macrophage membrane. Later stages of the phagocytosis are characterized by progressive and complete engulfment of both promastigotes and amastigotes. PMID- 10691340 TI - Nuclear industry family study:methods and description of a United Kingdom study linking occupational information held by employers to reproduction and child health. AB - OBJECTIVE: To describe the methods used in the nuclear industry family study for which a comprehensive database has been assembled that links employment in the nuclear industry and dosimetry records to information on employees' reproductive health and the health of their children. To discuss the response rates and characteristics of the study population. METHODS: Occupational cohort design leading to a retrospective cohort study of reproductive outcomes reported by 46 396 current and former employees of both sexes in the nuclear industry. Employees of nuclear establishments in the United Kingdom operated by the Atomic Energy Authority, the Atomic Weapons Establishment, and British Nuclear Fuels were surveyed with postal questionnaires ot collect information on pregnancies, children,and periods of infertility. Information on employment and monitoring for ionising radiation was supplied by the employing nuclear authority and was linked to pregnancies and periods of infertility with unique personal identification numbers. RESULTS: The design and completion of this study resulted in high quality data on a representative population of the Atomic Energy Authority, Atomic Weapons Establishment, and British Nuclear Fuels workforces. The response to the survey was extremely good (82% for male workers and 88% for female workers, excluding undelivered questionnaires), and a unique relational database has been created which will enable infertility, pregnancy, and child health outcomes to be examined with respect to the employment and radiation monitoring characteristics of parents. CONCLUSION: This is the first United Kingdom study to link detailed reproductive history data to occupational information held by employers. The methods developed for the study were found to be feasible and successful. The design can be adapted for other investigations of reproductive hazards to men and women in the workplace and is currently in use to survey over 100 000 armed forces personnel in an investigation of reproductive outcome among veterans of the Gulf war. PMID- 10691342 TI - Current awareness in geriatric psychiatry. PMID- 10691343 TI - Synthesis of (E)-9-(1-pyrenyl)-4-hydroxynon-2-enal, a fluorescent probe of the (E)-4-hydroxynon-2-enal with retained biological properties. AB - (E)-9-(1-pyrenyl)-4-hydroxynon-2-enal (FHNE), a fluorescent probe of (E)-4 hydroxynon-2-enal (HNE) is synthesised in seven steps and in 35% overall yield, starting from commercially available 1-pyrencarboxyaldehyde. When incubated with cultured HeLa cells this fluorescent probe penetrates cells and particularly concentrates in the region surrounding the nucleus. As the parent compound, HNE it is able to induce the activation of heat shock factor (HSF) and is able to induce the binding of HSF to heat shock element (HSE). PMID- 10691344 TI - Alar again: science, the media, and the public's right to know. PMID- 10691345 TI - Transitional cell carcinoma of the bladder in a patient on clean intermittent catheterization. PMID- 10691346 TI - Vegans, vegetarians and BPH. PMID- 10691347 TI - The pubofascial anchor sling procedure for recurrent genuine urinary stress incontinence. PMID- 10691348 TI - Extra-urethral urinary incontinence after incompetent vaginal obstetrics. PMID- 10691349 TI - Penile surgical techniques described by Oribasius (4th century CE) PMID- 10691350 TI - [Sampling studies in treatment follow-up. Evaluation of patient intake within the scope of comprehensive quality management at the Alt/Neuotting District Hospital]. PMID- 10691351 TI - [Treatment of mentally incompetent patients. Legal requirements in surgery]. PMID- 10691352 TI - [The current status of endoscopy in surgery]. PMID- 10691353 TI - [Continuous patient information during wound management]. PMID- 10691354 TI - [What is the outlook for January 1, 2000?]. PMID- 10691355 TI - [Distal radius fractures. II]. PMID- 10691356 TI - [Comment on B. Kreklau and G. Muhr: Preventive surgery on the musculoskeletal system]. PMID- 10691357 TI - [Comment on the position paper "Transplantation surgery in visceral surgery"]. PMID- 10691358 TI - [Deviation stoma. Comment on the contribution by H. R. Roosen and R. Schissel]. PMID- 10691359 TI - [The future of medicine at the threshold of the third millennium]. PMID- 10691360 TI - A less polluting pig. PMID- 10691361 TI - Protein protection from the sun. PMID- 10691362 TI - NRC: not enough data. PMID- 10691363 TI - Vitamin E vs. PCBs. PMID- 10691364 TI - Sins of the father. Parental smoking and childhood cancer. PMID- 10691365 TI - Catch the drift. Assessing risk from pesticide spraying. PMID- 10691366 TI - Malformed frogs. Making the leap to humans. PMID- 10691367 TI - The complement system has associations also with glucose metabolism. PMID- 10691368 TI - A pH-sensitive channel regulates urea access to Helicobacter pylori urease. PMID- 10691369 TI - C. difficile epidemic raises difficult questions about antibiotic-prescribing practices. PMID- 10691370 TI - Early negative results not the last word on secretin/autism story. PMID- 10691371 TI - Image of the month. Adherent, yellow exudate speckled with black spots in the distal two thirds of the esophagus. Diagnosis: This distinctive endoscopic image shows an acute necrotizing esophagitis, also known as the black esophagus. PMID- 10691372 TI - Abraham Vater of the ampulla (papilla) of Vater. PMID- 10691373 TI - Sensitivity of cold-preserved rat liver to single-pass reperfusion. PMID- 10691374 TI - H. pylori and risk of ulcer bleeding among users of NSAIDS. PMID- 10691375 TI - Enterokinase in cerulein pancreatitis. PMID- 10691376 TI - Lack of effect of acid suppression therapy on gastric atrophy. PMID- 10691377 TI - Role of the CDP-choline and the alternative pathway for phosphatidylcholine biosynthesis. PMID- 10691378 TI - Amino acid sequence of EC2 domain in CD81 is highly conserved in Japanese subjects. PMID- 10691379 TI - Lamivudine treatment for children with interferon refractory chronic hepatitis B. PMID- 10691380 TI - Herbal products for liver diseases. PMID- 10691381 TI - Inhibition of the cyclooxygenase/lipoxygenase pathways to improve interferon alfa efficacy in chronic hepatitis C: a wrong track. PMID- 10691382 TI - Immune therapy of hepatitis B virus chronic infection. PMID- 10691383 TI - Iron and fibrosis in nonalcoholic fatty liver disease. PMID- 10691384 TI - Gastroenterology on the Internet--I. PMID- 10691385 TI - Lung volume reduction for patients with severe COPD. PMID- 10691386 TI - Selective decontamination of the digestive tract in critically ill patients. PMID- 10691387 TI - Decompressive craniectomy to treat intracranial hypertension in head injury patients. PMID- 10691388 TI - Alcohol, thiamin and fibromyalgia. PMID- 10691389 TI - The power of grassroots commitment. PMID- 10691391 TI - Eight faces of validity. PMID- 10691390 TI - Folic acid and Down syndrome. PMID- 10691392 TI - Surgical-site complications associated with a morphine nerve paste used for postoperative pain control after laminectomy. PMID- 10691393 TI - Proceedings of the 4th International Conference of the Hospital Infection Society. Edinburgh, 13-17 September 1998. PMID- 10691394 TI - Risk aversion and costs: a comparison of family physicians and general internists. PMID- 10691395 TI - [Resection of osteoid osteoma]. PMID- 10691396 TI - [Clinical medical surveillance strategy of persons exposed to asbestos. French Occupational Health Society, Society of French-speaking Pneumologists, Society of Thoracic Imaging]. PMID- 10691397 TI - [Case no.1. Diagnosis: Balo concentric sclerosis]. PMID- 10691398 TI - [Case no. 2. Congenital portocaval anastomosis]. PMID- 10691399 TI - [Case no.3. Diagnosis: cervical vertebral calcinosis associated with systemic scleroderma]. PMID- 10691400 TI - [Case no. 4. Subdural sarcoma]. PMID- 10691402 TI - [Case no. 1. Diagnosis: mucoepidermoid carcinoma of the left bronchus]. PMID- 10691401 TI - [Case no. 5. Diagnosis: dural arteriovenous fistula of the posterior fossa with perimedullary venous drainage]. PMID- 10691403 TI - [Case no. 2. Diagnosis: ileal, retroperitoneal and pulmonary metastases from a malignant melanoma]. PMID- 10691404 TI - [Case no. 3. Diagnosis: urinoma]. PMID- 10691405 TI - [Case no. 4. Diagnosis: bilateral breast lymphoma type B]. PMID- 10691406 TI - [Case no. 5. Diagnosis: disseminated pulmonary and medullary tuberculosis in chronic renal insufficiency with pathologic signs associated with secondary hyperparathyroidism]. PMID- 10691407 TI - [Case no. 6. Diagnosis: multiple endocrine neoplasia type 1]. PMID- 10691408 TI - [Case no. 7. Diagnosis: acute retrococecal appendicitis]. PMID- 10691409 TI - [Case no. 8. Diagnosis: esophageal metastasis from treated breast cancer (infiltrating lobular type)]. PMID- 10691410 TI - Risk of multisystem disease in isolated ocular angioma (haemangioblastoma) PMID- 10691411 TI - Confirmation of the assignment of the Sanjad-Sakati (congenital hypoparathyroidism) syndrome (OMIM 241410) locus to chromosome lq42-43. PMID- 10691412 TI - Molecular diagnosis is important to confirm suspected pseudoachondroplasia. PMID- 10691413 TI - Biallelic expression of IGFBP1 and IGFBP3, two candidate genes for the Silver Russell syndrome. PMID- 10691414 TI - Mutational analysis of the human pancreatic secretory trypsin inhibitor (PSTI) gene in hereditary and sporadic chronic pancreatitis. PMID- 10691415 TI - A case of inv dup(8p) with early onset breast cancer. PMID- 10691416 TI - Appendiceal carcinoma complicating adenomatous polyposis in a young woman with a de novo constitutional reciprocal translocation t(5;8)(q22;p23.1) PMID- 10691417 TI - NF2 gene deletion in a family with a mild phenotype. PMID- 10691418 TI - Absence of fragile X syndrome in Nova Scotia. PMID- 10691419 TI - The mystery of addiction. PMID- 10691420 TI - The globalization of Chinese medicine. PMID- 10691421 TI - The development of dentistry, 1000-2000. PMID- 10691422 TI - Crowning achievements in dentistry. PMID- 10691423 TI - Diagnosis: the doctor and the urine glass. PMID- 10691424 TI - Medical diagnosis in the Internet age. PMID- 10691425 TI - Disease and stigma. PMID- 10691426 TI - The "geneticisation" of disease stigma. PMID- 10691427 TI - Doctors' status: changes in the past millennium. PMID- 10691428 TI - Status of the doctor--present and future. PMID- 10691429 TI - Economic development and disease. PMID- 10691430 TI - Addiction and the truth in magic realism. PMID- 10691431 TI - Economics and health. PMID- 10691432 TI - Early observations of genetic diseases. PMID- 10691433 TI - Gene therapy and beyond. PMID- 10691434 TI - Magic and the origins of modern science. PMID- 10691435 TI - Traditional healing practice using medicinal herbs. PMID- 10691436 TI - Health care in hospitals: the past 1000 years. PMID- 10691437 TI - Hospitals in developing countries: a weak link in a weak chain. PMID- 10691438 TI - Child mortality: social and medical responses. PMID- 10691439 TI - Evidence-based and value-based paediatrics. PMID- 10691440 TI - Influenza--a continuing detective story. PMID- 10691441 TI - Ageing: old visions, new times? PMID- 10691442 TI - Prevention and control of influenza. PMID- 10691443 TI - The long road to malaria eradication. PMID- 10691444 TI - Antimalarial drug policy: making systematic change. PMID- 10691445 TI - The history of medical teaching. PMID- 10691446 TI - Medical education for the doctors of tomorrow. PMID- 10691447 TI - Nine centuries of cranial surgery. PMID- 10691448 TI - The next 100 years of neurosurgery. PMID- 10691449 TI - The Potato Eaters. PMID- 10691450 TI - Food and nutrition for all. PMID- 10691451 TI - A millennium of obstetrics and gynaecology. PMID- 10691452 TI - Ageing in the 21st century. PMID- 10691453 TI - The brave new world of making babies. PMID- 10691454 TI - Origin and evolution of medical oncology. PMID- 10691455 TI - The future of oncology: more of the same? PMID- 10691456 TI - Old news in ophthalmology. PMID- 10691457 TI - A mission for vision. PMID- 10691458 TI - "Orthopaedics"--before and after the word. PMID- 10691459 TI - Computer-assisted tools and interventional technologies. PMID- 10691461 TI - Today's patient: passive or involved? PMID- 10691460 TI - Subjectivity and ethics: the patient. PMID- 10691462 TI - Perception of disease and its meanings. PMID- 10691463 TI - The hope of anaesthesia. PMID- 10691464 TI - The concept of disease: from Newton back to Aristotle. PMID- 10691465 TI - "Pharmacocentricity": from elixirs to magic bullets. PMID- 10691466 TI - New directions in pharmacology. PMID- 10691468 TI - Can plagues be predicted, prevented? PMID- 10691467 TI - Poisoners and "plague-smearers". PMID- 10691469 TI - The "alter ego" in psychiatry. PMID- 10691470 TI - Changing boundaries in psychiatry. PMID- 10691471 TI - Transformations in social medicine. PMID- 10691473 TI - Trials and errors in clinical research. PMID- 10691472 TI - Public health: the outlook for contrasting populations. PMID- 10691474 TI - The two pillars of anaesthesia. PMID- 10691475 TI - Biomedical research: crossing the 1 s Rubicon. PMID- 10691476 TI - The wages of sin. PMID- 10691477 TI - The staying power of sexually transmitted diseases. PMID- 10691478 TI - Pandora's box opened: 1000 years of war and disease. PMID- 10691479 TI - Weapons for the future. PMID- 10691480 TI - Women and doctors in medicine. PMID- 10691481 TI - Finale. PMID- 10691482 TI - The changing heart. PMID- 10691483 TI - Cardiology: where to go from here? PMID- 10691484 TI - The past 1000 years of Chinese medicine. PMID- 10691485 TI - Domestic violence. PMID- 10691486 TI - Domestic violence. PMID- 10691487 TI - Domestic violence. PMID- 10691488 TI - Domestic violence. PMID- 10691489 TI - The long-QT syndrome. PMID- 10691490 TI - AIDS in the 21st century. PMID- 10691491 TI - AIDS in the 21st century. PMID- 10691492 TI - Racial differences in the treatment of early-stage lung cancer. PMID- 10691493 TI - Racial differences in the treatment of early-stage lung cancer. PMID- 10691494 TI - Racial differences in the treatment of early-stage lung cancer. PMID- 10691495 TI - The unintended consequences of measuring quality on the quality of medical care. PMID- 10691496 TI - The unintended consequences of measuring quality on the quality of medical care. PMID- 10691497 TI - The unintended consequences of measuring quality on the quality of medical care. PMID- 10691498 TI - Diagnosis of scombroid poisoning by measurement of plasma histamine. PMID- 10691499 TI - Disclosure of authors' conflicts of interest: a follow-up. PMID- 10691500 TI - Behcet's disease. PMID- 10691501 TI - Behcet's disease. PMID- 10691502 TI - Behcet's disease. PMID- 10691503 TI - Behcet's disease. PMID- 10691504 TI - Behcet's disease. PMID- 10691505 TI - Behcet's disease. PMID- 10691506 TI - Absence of benefit of eradicating Helicobacter pylori in patients with nonulcer dyspepsia. PMID- 10691507 TI - Absence of benefit of eradicating Helicobacter pylori in patients with nonulcer dyspepsia. PMID- 10691508 TI - Electrocardiographic artifact. PMID- 10691509 TI - Electrocardiographic artifact. PMID- 10691510 TI - Electrocardiographic artifact. PMID- 10691511 TI - Electrocardiographic artifact. PMID- 10691512 TI - Lamivudine for the treatment of chronic hepatitis B. PMID- 10691513 TI - Myocardial infarction in a patient with hypertrophic cardiomyopathy. PMID- 10691514 TI - Canadian biomedical collaboration keeps on growing. PMID- 10691515 TI - [Accidental fall of an infant from a baby carrier]. PMID- 10691516 TI - Hemolytic uremic syndrome associated with invasive Streptococcus pneumoniae infection. PMID- 10691517 TI - Urinary tract infection guidelines questioned. PMID- 10691518 TI - Urinary tract infection guidelines questioned. PMID- 10691519 TI - Urinary tract infection guidelines questioned. PMID- 10691520 TI - Urinary tract infection guidelines questioned. PMID- 10691521 TI - Benign enlargement of the mandibulofacial lymph node. PMID- 10691522 TI - Response to the joint statement on HIV screening. PMID- 10691524 TI - Programmed proteolysis and the control of cell division. A discussion meeting, November 4-5, 1998. PMID- 10691523 TI - Article captures the essence and meaning of alopecia. PMID- 10691525 TI - 21st Annual Winter Neuropeptide Conference. Breckenridge, Colorado, USA. January 29-February 1, 2000. Abstracts. PMID- 10691526 TI - 2001 budget. Clinton seeks 'major lift' in U.S. research programs. PMID- 10691527 TI - Patents. Company gets rights to cloned human embryos. PMID- 10691528 TI - Cell biology. Generating new yeast prions. PMID- 10691529 TI - Scientific publishing. Publishers discuss European e-print site. PMID- 10691530 TI - Restoration ecology. Bringing the Salton Sea back to life. PMID- 10691531 TI - FDA halts all gene therapy trials at Penn. PMID- 10691532 TI - Intellectual property. NIH cuts deal on use of OncoMouse. PMID- 10691533 TI - Pasteur Institute. New chief promises renewal and openness. PMID- 10691534 TI - Meeting. Society for Integrative and Comparative Biology. An integrative science finds a home. PMID- 10691535 TI - Restoration ecology. Returning America's forests to their 'natural' roots. PMID- 10691536 TI - Unseemly competition. PMID- 10691537 TI - E-knowledge hullabaloo--or when will the glass spill over? PMID- 10691538 TI - Gene therapy on trial. PMID- 10691539 TI - Sickle cell anemia therapy: progress since Pauling. PMID- 10691540 TI - Oversight mechanisms for clinical research. PMID- 10691541 TI - Perspectives: epidemiology. Simple rules with complex dynamics. PMID- 10691542 TI - HIV infection and dementia. PMID- 10691543 TI - Perspectives: earth science and evolution. Genomics and the geosciences. PMID- 10691544 TI - 2001 budget. How NSF came up with the biggest boost in history. PMID- 10691545 TI - Biotechnology. Both sides claim victory in trade pact. PMID- 10691546 TI - A face-off over tumor blood supply. PMID- 10691547 TI - Plant biotechnology. Consumer power heralds hard times for researchers. PMID- 10691548 TI - Neuroscience. Cold numbers unmake the quantum mind. PMID- 10691549 TI - Ecology. How climate change alters rhythms of the wild. PMID- 10691550 TI - Lunch selections expanding. PMID- 10691551 TI - Antibiotic rotation. PMID- 10691552 TI - Herbicide use on roundup ready crops. PMID- 10691553 TI - Benefits of membership. PMID- 10691554 TI - From Turin to Stockholm via St. Louis and Rio de Janeiro. PMID- 10691555 TI - A life in science. PMID- 10691556 TI - Perspectives: neurobiology. Regeneration in the Nogo zone. PMID- 10691557 TI - Perspectives: drug delivery. Regulating export of ER cargo. PMID- 10691558 TI - Perspectives: behavior. Measuring beelines to food. PMID- 10691559 TI - Perspectives: biosynthetic pathways. Biosynthesis meets bioinformatics. PMID- 10691560 TI - HIV transmission. AIDS researchers look to Africa for new insights. PMID- 10691561 TI - Scientific advice. Academies get together to tackle the big issues. PMID- 10691562 TI - Neurobiology. A new clue to how alcohol damages brains. PMID- 10691563 TI - Stem cells. Wisconsin to distribute embryonic cell lines. PMID- 10691564 TI - Stem cells. Report would open up research in Japan. PMID- 10691565 TI - Human genetics. Start-up claims piece of Iceland gene pie. PMID- 10691567 TI - Plan to reduce number of new grants tempers enthusiasm for NIH budget hike. PMID- 10691566 TI - Balancing the science budget. PMID- 10691568 TI - AIDS research. Vaccine studies stymied by shortage of animals. PMID- 10691569 TI - Veterinary perspective on gene therapy with adenoviruses. PMID- 10691570 TI - Luzia is not alone. PMID- 10691571 TI - Economics of bushmeat. PMID- 10691572 TI - Perspectives: signal transduction. Signals to move cells. PMID- 10691573 TI - Perspectives: evolution. Is bigger better in cricket? PMID- 10691574 TI - Proceedings of the 11th Japanese-German Cooperative Symposium on Protozoan Diseases. August 29-31, 1998. PMID- 10691575 TI - Morphology of Toxoplasma tachyzoites in the brain of an AIDS patient. PMID- 10691576 TI - [Advance directives: words pass, writing remains?]. PMID- 10691577 TI - [Tumescence local anesthesia. Interview with Prof. Dr. Werner Mang on the developmental state of this new local anesthesia method. Interview by Werner Rossling/Hinrich Kuster]. PMID- 10691578 TI - [Gene therapy. Basis and preliminary therapy concept for the treatment of malignant diseases of the urogenital system]. PMID- 10691579 TI - [Therapy of common PCA (acinar adenocarcinoma). German Society of Urology]. PMID- 10691580 TI - [Research support for urology. A guideline--II. German Society of Urology]. PMID- 10691581 TI - [Aleksandr Vasil'evich Mel'nikov (1889-1958)]. PMID- 10691582 TI - Inhaled corticosteroids? PMID- 10691583 TI - Dizziness among older adults: a possible geriatric syndrome. AB - BACKGROUND: In previous studies of dizziness, the prevalence of specific causes has varied widely and either no or multiple causes have been identified. Dizziness might be better considered a geriatric syndrome that results from impairment or disease in multiple systems. OBJECTIVE: To determine the predisposing characteristics and situational factors associated with dizziness. DESIGN: Population-based, cross-sectional study. SETTING: Community. PARTICIPANTS: Probability sample of 1087 community-living persons in New Haven, Connecticut, who were at least 72 years of age. MEASUREMENTS: Episodes of dizziness that occurred for at least 1 month; manifestations of dizziness; and predisposing demographic, medical, neurologic, sensory, and psychological characteristics. RESULTS: 261 participants (24%) reported dizziness; 56% of dizzy persons described several sensations and 74% reported several triggering activities. The adjusted relative risks for characteristics associated with dizziness were 1.69 (95% CI, 1.24 to 2.30) for anxiety, 1.36 (CI, 1.02 to 1.80) for depressive symptoms, 1.27 (CI, 0.99 to 1.63) for impaired hearing, 1.30 (CI, 1.01 to 1.68) for five or more medications, 1.31 (CI, 0.92 to 1.87) for postural hypotension, 1.34 (CI, 0.95 to 1.90) for impaired balance, and 1.31 (CI, 1.00 to 1.71) for past myocardial infarction. The adjusted relative risk for dizziness was 1.38 (CI, 1.27 to 1.49) for each additional characteristic. CONCLUSIONS: The association among characteristics in multiple domains (cardiovascular, neurologic, sensory, psychological, and medication-related) and dizziness, coupled with the multiplicity of sensations and triggering activities, suggests that dizziness may be a geriatric syndrome, similar to delirium and falling. If so, an impairment reduction strategy, proven effective for other geriatric syndromes, may be effective in reducing the symptoms and disabilities associated with dizziness. PMID- 10691584 TI - Low fractional calcium absorption increases the risk for hip fracture in women with low calcium intake. Study of Osteoporotic Fractures Research Group. AB - BACKGROUND: Decreased ability to absorb calcium with age limits adaptation to low calcium intake and is thought to lead to secondary hyperparathyroidism and increased risk for hip and other fractures. However, the associations between fractional calcium absorption, dietary calcium intake, and risk for fracture have never been studied. OBJECTIVE: To determine whether low fractional calcium absorption in women with low calcium intake increases the risk for subsequent hip and other nonspine fractures. DESIGN: Prospective cohort study. SETTING: Four clinical centers in Baltimore County, Maryland; Portland, Oregon; Minneapolis, Minnesota; and the Monongahela Valley, Pennsylvania. PARTICIPANTS: 5452 nonblack women 69 years of age or older participating in the fourth examination of the Study of Osteoporotic Fractures. MEASUREMENTS: Fractional calcium absorption was measured by using a 3-hour single isotope (45Ca) technique. Incident fractures were identified prospectively and were confirmed by radiographic report. RESULTS: During an average of 4.8 years, 729 women (13%) experienced at least one nonspine fracture; 153 of these women had hip fractures. After adjustment for age, women with lower fractional calcium absorption were at increased risk for hip fracture (relative risk per 1-SD [7.7%] decrease in fractional calcium absorption, 1.24 [95% CI, 1.05 to 1.48]). Women with low fractional calcium absorption and low calcium intake were at greatest risk for subsequent hip fracture; among women whose dietary calcium intake was less than 400 mg/d, those who had fractional calcium absorption at or below the median value of 32.3% had a 2.5-fold (CI, 1.29 fold to 4.69-fold) increase in risk for hip fracture compared with those who had greater absorption efficiency. Fractional calcium absorption was not related to risk for other nonspine fractures (relative risk per 1-SD [7.7%] decrease in fractional calcium absorption, 1.05 [CI, 0.96 to 1.14]). CONCLUSIONS: In elderly women, low fractional calcium absorption in the setting of low calcium intake increases the risk for hip fracture. Our findings support the hypothesis of type II osteoporosis, which postulates that decreased calcium absorption is an important risk factor for hip fracture in older persons. PMID- 10691585 TI - Effects of fexofenadine, diphenhydramine, and alcohol on driving performance. A randomized, placebo-controlled trial in the Iowa driving simulator. AB - BACKGROUND: Sedating antihistamines may impair driving performance as seriously as alcohol. OBJECTIVE: To compare the effects of fexofenadine, diphenhydramine, alcohol, and placebo on driving performance. DESIGN: Randomized, double-blind, double-dummy, four-treatment, four-period crossover trial. SETTING: The Iowa Driving Simulator. PARTICIPANTS: 40 licensed drivers with seasonal allergic rhinitis who were 25 to 44 years of age. INTERVENTION: One dose of fexofenadine (60 mg), diphenhydramine (50 mg), alcohol (approximately 0.1% blood alcohol concentration), or placebo, given at weekly intervals before participants drove for 1 hour in the Iowa Driving Simulator. MEASUREMENTS: The primary end point was coherence, a continuous measure of participants' ability to match the varying speed of a vehicle that they were following. Secondary end points were drowsiness and other driving measures, including lane keeping and response to a vehicle that unexpectedly blocked the lane ahead. RESULTS: Participants had significantly better coherence after taking alcohol or fexofenadine than after taking diphenhydramine. Lane keeping (steering instability and crossing the center line) was impaired after alcohol and diphenhydramine use compared with fexofenadine use. Mean response time to the blocking vehicle was slowest after alcohol use (2.21 seconds) compared with fexofenadine use (1.95 seconds). Self-reported drowsiness did not predict lack of coherence and was weakly associated with minimum following distance, steering instability, and leftlane excursion. CONCLUSIONS: Participants had similar performance when treated with fexofenadine or placebo. After alcohol use, participants performed the primary task well but not the secondary tasks; as a result, overall driving performance was poorer. After participants took diphenhydramine, driving performance was poorest, indicating that diphenhydramine had a greater impact on driving than alcohol did. Drowsiness ratings were not a good predictor of impairment, suggesting that drivers cannot use drowsiness to indicate when they should not drive. PMID- 10691586 TI - Recombinant human thrombopoietin attenuates carboplatin-induced severe thrombocytopenia and the need for platelet transfusions in patients with gynecologic cancer. AB - BACKGROUND: Thrombocytopenia is a significant problem in the treatment of cancer. OBJECTIVE: To assess the clinical safety of therapy with recombinant human thrombopoietin (rhTPO) and its ability to ameliorate chemotherapy-induced severe thrombocytopenia. DESIGN: Phase I/II clinical cohort study. SETTING: The University of Texas M.D. Anderson Cancer Center, Houston, Texas. PATIENTS: 29 patients with gynecologic cancer. INTERVENTION: Recombinant human thrombopoietin was given before chemotherapy and after a second cycle of carboplatin therapy. MEASUREMENTS: Peripheral blood counts and platelet transfusions. RESULTS: Administration of rhTPO after chemotherapy significantly reduced the degree and duration of thrombocytopenia and enhanced platelet recovery. In patients who received the optimal biological dose of rhTPO (1.2 microg/kg of body weight) in cycle 2 (carboplatin plus rhTPO), the mean platelet count nadir was higher (44x10(9) cells/L and 20x10(9) cells/L; P = 0.002) and the duration of thrombocytopenia was shorter (days with a platelet count <20x10(9) cells/L, 1 and 4 [P = 0.002]; days with a platelet count <50x10(9) cells/L, 4 and 7 [P = 0.006]) than in cycle 1 (carboplatin only). The need for platelet transfusion in this group was reduced from 75% of patients in cycle 1 to 25% of patients in cycle 2 (P = 0.013). CONCLUSIONS: Therapy with rhTPO seems to be safe and may attenuate chemotherapy-induced severe thrombocytopenia and reduce the need for platelet transfusions. PMID- 10691587 TI - Increased susceptibility to pulmonary emphysema among HIV-seropositive smokers. AB - BACKGROUND: Previous uncontrolled reports have suggested that HIV-seropositive persons develop an accelerated form of emphysema. OBJECTIVE: To characterize the risk for emphysema in a stable HIV-seropositive outpatient population. DESIGN: Controlled, cross-sectional analysis. SETTING: Midwestern urban community. PARTICIPANTS: HIV-seropositive persons (n = 114) without AIDS-related pulmonary complications and HIV-seronegative controls (n = 44), matched for age and smoking history. MEASUREMENTS: Measurement of pulmonary function, bronchoalveolar lavage, and high-resolution computed tomography of the chest. RESULTS: The incidence of emphysema was 15% (17 of 114) in the HIV-seropositive group compared with 2% (1 of 44) in the HIV-seronegative group (P = 0.025). The incidence of emphysema in participants with a smoking history of 12 pack-years or greater was 37% (14 of 38 persons) in the HIV-seropositive group compared with 0% (0 of 14 persons) in the HIV-seronegative group (P = 0.011). The percentage of cytotoxic lymphocytes in lavage fluid was much higher in HIV-seropositive smokers with emphysema. CONCLUSIONS: Infection with HIV accelerates the onset of smoking-induced emphysema. The results of this study support the emerging concept that cytotoxic lymphocytes may have an important role in emphysema pathogenesis. PMID- 10691588 TI - Developing and implementing computerized protocols for standardization of clinical decisions. AB - Humans have only a limited ability to incorporate information in decision making. In certain situations, the mismatch between this limitation and the availability of extensive information contributes to the varying performance and high error rate of clinical decision makers. Variation in clinical practice is due in part to clinicians' poor compliance with guidelines and recommended therapies. The use of decision-support tools is a response to both the information revolution and poor compliance. Computerized protocols used to deliver decision support can be configured to contain much more detail than textual guidelines or paper-based flow diagrams. Such protocols can generate patient-specific instructions for therapy that can be carried out with little interclinician variability; however, clinicians must be willing to modify personal styles of clinical management. Protocols need not be perfect. Several defensible and reasonable approaches are available for clinical problems. However, one of these reasonable approaches must be chosen and incorporated into the protocol to promote consistent clinical decisions. This reasoning is the basis of an explicit method of decision support that allows the rigorous evaluation of interventions, including use of the protocols themselves. Computerized protocols for mechanical ventilation and management of intravenous fluid and hemodynamic factors in patients with the acute respiratory distress syndrome provide case studies for this discussion. PMID- 10691589 TI - Update in hospital medicine. PMID- 10691590 TI - Prevention of intravascular catheter-related infections. AB - PURPOSE: To review the literature on prevention of intravascular catheter-related infections. DATA SOURCES: The MEDLINE database, conference proceedings, and bibliographies of review articles and book chapters were searched for relevant articles. Primary authors were contacted directly if data were incomplete. STUDY SELECTION: Studies met the following criteria unless otherwise stated: Trials were prospective and randomized; catheters were inserted into new sites, not into old sites over guidewires; catheter cultures were done by using semi-quantitative or quantitative methods; and, for prospective studies, catheter-related bloodstream infection was confirmed by microbial growth from percutaneously drawn blood cultures that matched catheter cultures. DATA EXTRACTION: Data on population, methods, preventive strategy, and outcome (measured as catheter related bloodstream infections) were gathered. The quality of the data was graded by using preestablished criteria. DATA SYNTHESIS: The recommended preventive strategies with the strongest supportive evidence are full barrier precautions during central venous catheter insertion; subcutaneous tunneling short-term catheters inserted in the internal jugular or femoral veins when catheters are not used for drawing blood; contamination shields for pulmonary artery catheters; povidone-iodine ointment applied to insertion sites of hemodialysis catheters; specialized nursing teams caring for patients with short-term peripheral venous catheters, especially at institutions with a high incidence of catheter-related infection; no routine replacement of central venous catheters; antiseptic chamberfilled hub or hub-protective antiseptic sponge for central venous catheters; and use of chlorhexidine-silver sulfadiazine-impregnated or minocycline-rifampin-impregnated short-term central venous catheters if the rate of infection is high despite adherence to other strategies that do not incorporate antimicrobial agents (for example, maximal barrier precautions). CONCLUSIONS: Simple interventions can reduce the risk for serious catheter related infection. Adequately powered randomized trials are needed. PMID- 10691591 TI - Occam's razor, geriatric syndromes, and the dizzy patient. PMID- 10691592 TI - Nonsedating antihistamines should be preferred over sedating antihistamines in patients who drive. PMID- 10691593 TI - Responding to intractable terminal suffering: the role of terminal sedation and voluntary refusal of food and fluids. ACP-ASIM End-of-Life Care Consensus Panel. American College of Physicians-American Society of Internal Medicine. AB - When provided by a skilled, multidisciplinary team, palliative care is highly effective at addressing the physical, psychological, social, and spiritual needs of dying patients and their families. However, some patients who have witnessed harsh death want reassurance that they can escape if their suffering becomes intolerable. In addition, a small percentage of terminally ill patients receiving comprehensive care reach a point at which their suffering becomes severe and unacceptable despite unrestrained palliative efforts; some of these patients request that death be hastened. This paper presents terminal sedation and voluntary refusal of hydration and nutrition as potential last resorts that can be used to address the needs of such patients. These two practices allow clinicians to address a much wider range of intractable end-of-life suffering than physician-assisted suicide (even if it were legal) and can also provide alternatives for patients, families, and clinicians who are morally opposed to physician-assisted suicide. This paper will define the two practices, distinguish them from more standard palliative care interventions and from physician-assisted suicide, illustrate them with a real clinical scenario, provide potential guidelines and practicalities, and explore their moral and legal status. Although medicine cannot sanitize dying or provide perfect answers for all challenging end of-life clinical problems, terminal sedation and voluntary refusal of hydration and nutrition substantially increase patients' choices at this inherently challenging time. PMID- 10691594 TI - Trends in tuberculosis transmission. PMID- 10691595 TI - Mycophenolate mofetil-induced dyshidrotic eczema. PMID- 10691596 TI - Venlafaxine-associated hepatitis. PMID- 10691597 TI - Hydroxyurea-induced leg ulcers treated with Apligraf. PMID- 10691598 TI - Importance of proper identification of stinging insects. PMID- 10691599 TI - Understanding adherence to HIV medication. PMID- 10691600 TI - Use of statins and fibrates in hyperlipidemic patients with neuromuscular disorders. PMID- 10691602 TI - Editor's change of address PMID- 10691601 TI - Richard Lower: anatomist and physiologist. PMID- 10691603 TI - Biotransformation of beta-myrcene by the larvae of common cutworm (Spodoptera litura). AB - beta-Myrcene was mixed in an artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. beta-Myrcene was transformed mainly to myrcene-3(10)-glycol and myrcene-1,2-glycol. Each pair of double bonds of beta-myrcene was converted to the corresponding diol by oxidation, respectively. The 3,10- and 1,2-double bonds of beta-myrcene were respectively oxidized. PMID- 10691604 TI - Determination of polycyclic aromatic hydrocarbons in commercial liquid smoke flavorings of different compositions by gas chromatography-mass spectrometry. AB - The presence of polycyclic aromatic hydrocarbons (PAHs) in five commercial liquid smoke flavorings, used in the European food industry, was studied. The samples were subjected to an alkaline treatment, extracted with cyclohexane, cleaned up by means of solid-phase extraction tubes, and analyzed by gas chromatography-mass spectrometry. Three different procedures for the cleanup were tested. The results revealed the presence of 34 PAHs, some of them with methyl substituents. In all cases, the concentrations of compounds of low molecular weight were much higher than those of high molecular weight. Relationships between smoke flavoring compositions and PAH levels were also studied. Three of the samples contained high levels of both total and carcinogenic PAHs. Benzo[a]pyrene was also detected in these three samples, but its concentration did not exceed the 10 microg/kg level fixed by the FAO/WHO. Finally, a relation was found, first between the concentrations of total carcinogenic PAHs and benzo[a]pyrene and also between the concentrations of pyrene and benzo[a]pyrene. The latter ratio reveals that pyrene concentration could be very useful in predicting the level of benzo[a]pyrene and, consequently, in estimating the carcinogenicity arising from the presence of benzo[a]pyrene and other carcinogenic PAHs. PMID- 10691605 TI - Product and redox potential analysis of sauerkraut fermentation. AB - The relationships between the redox potential of the brine, during fermentation of white cabbage into sauerkraut of two early and two late fermentation processes, and the changes in the amount of sugars, organic acids, the redox potential of the brine and of the ascorbic acid redox couple, and pH are described. The trend in the change of the redox potential of the brine is the same for all four fermentation processes studied. In the first phase a sharp decrease in redox potential is followed by an increase in redox potential. In the second phase the redox potential is rather constant. This second phase is followed by another decrease in redox potential, which stabilizes at a minimum value, the third phase. It was observed that sugar fermentation and acid production mainly took place during the first and third phases, probably representing, respectively, the heterogeneous and homogeneous fermentation processes. PMID- 10691606 TI - Antioxidant activity in fruits and leaves of blackberry, raspberry, and strawberry varies with cultivar and developmental stage. AB - Fruits and leaves from different cultivars of thornless blackberry (Rubus sp.), red raspberry (Rubus idaeus L.), black raspberry (Rubus occidentalis L.), and strawberry (Fragaria x ananassa D.) plants were analyzed for total antioxidant capacity (oxygen radical absorbance capacity, ORAC) and total phenolic content. In addition, fruits were analyzed for total anthocyanin content. Blackberries and strawberries had the highest ORAC values during the green stages, whereas red raspberries had the highest ORAC activity at the ripe stage. Total anthocyanin content increased with maturity for all three species of fruits. Compared with fruits, leaves were found to have higher ORAC values. In fruits, ORAC values ranged from 7.8 to 33.7 micromol of Trolox equivalents (TE)/g of fresh berries (35. 0-162.1 micromol of TE/g of dry matter), whereas in leaves, ORAC values ranged from 69.7 to 182.2 micromol of TE/g of fresh leaves (205.0-728.8 micromol of TE/g of dry matter). As the leaves become older, the ORAC values and total phenolic contents decreased. The results showed a linear correlation between total phenolic content and ORAC activity for fruits and leaves. For ripe berries, a linear relationship existed between ORAC values and anthocyanin content. Of the ripe fruits tested, on the basis of wet weight of fruit, cv. Jewel black raspberry and blackberries may be the richest source for antioxidants. On the basis of the dry weight of fruit, strawberries had the highest ORAC activity followed by black raspberries (cv. Jewel), blackberries, and red raspberries. PMID- 10691607 TI - Low-density lipoprotein antioxidant activity of phenolic compounds and polyphenol oxidase activity in selected clingstone peach cultivars. AB - The antioxidant potential of eight clingstone peach cultivars was investigated by determining phenolic compounds and inhibition of low-density lipoprotein (LDL) oxidation. Cultivars low in polyphenol oxidase (PPO) were also selected to minimize enzymatic browning. Inhibition of LDL oxidation varied from 17.0 to 37.1% in peach flesh extract, from 15.2 to 49.8% in whole peach extract, and from 18.2 to 48.1% in peel extract. Total phenols were 432.8-768.1 mg/kg in flesh extract, 483.3-803.0 mg/kg in whole extract, and 910.9-1922.9 mg/kg in peel extract. The correlation coefficient between relative LDL antioxidant activity and concentration of total phenols was 0.76. Peel PPO activity was higher than flesh activity in most cultivars. The lowest PPO and specific activities were found in the Walgant cultivar, followed by Kakamas and 18-8-23. These three cultivars combine the desirable characteristics of strong antioxidant activity, low PPO activity, and lower susceptibility to browning reactions. PMID- 10691608 TI - Transglycosylation of neohesperidin dihydrochalcone by Bacillus stearothermophilus maltogenic amylase. AB - Neohesperidin dihydrochalcone (NHDC), a sweet compound derived from citrus fruits, was modified to a series of its oligosaccharides by transglycosylation activity of Bacillus stearothermophilus maltogenic amylase (BSMA). Maltotriose as a donor was reacted with NHDC as an acceptor to glycosylate for the purpose of increasing the solubility of NHDC. Maltosyl-NHDC was a major transglycosylation product among the several transfer products by TLC analysis. The structure of the major transglycosylation product was determined to be maltosyl-alpha-(1,6) neohesperidin dihydrochalcone by MALDI-TOF/MS and (1)H and (13)C NMR. Maltosyl NHDC was 700 times more soluble in water and 7 times less sweet than NHDC. PMID- 10691609 TI - Selectivity of celite-immobilized patatin (lipid acyl hydrolase) from potato (Solanum tuberosum L.) tubers in esterification reactions As influenced by water activity and glycerol analogues as alcohol acceptors. AB - Lipid acyl hydrolase (LAH; patatin) was purified from potato tubers by ammonium sulfate fractionation followed by anion-exchange and affinity chromatography. The major protein band of 40-43 kDa on SDS-PAGE appeared to be patatin, and it stained positive for lipase activity on native PAGE. Selectivity of a Celite immobilized potato LAH in esterification reactions with n-acyl fatty acids (FA; C4, C6, C8, C10, C12, C14, C16, and C18) and alcohol acceptors (n-propanol, 2 propanol, 1,3-propanediol, and glycerol; 1,2-propanediol was not sufficiently reactive) was studied in isooctane. Immobilized LAH was highly selective for medium chain FAs (C8/C10) with a secondary optimum for chain lengths of C14/16. Water activity (a(w)) influenced activity and FA selectivity of the enzyme. Initial rates of ester synthesis were greatest at a(w) of 0.90 for all alcohol acceptors except for glycerol, where greatest initial rates were observed at a(w) of 0.19. Immobilized LAH preparations exhibited a bell-shape pH profile with optimum activity at pH 6-7 for ester synthesis, and no effect of pH on FA selectivity was observed. PMID- 10691610 TI - Rapid separation of lysozyme from chicken egg white by reductants and thermal treatment. AB - Reductants (0.1-2.0% ascorbic acid, cysteine, or cystine and 0.04-1. 0% beta mercaptoethanol) were added to 5-fold diluted, salted duck egg whites (commercially and laboratory prepared) and fresh egg whites (chicken and duck), and subsequently the mixtures were heated at 70 degrees C for 1-10 min. The maximal recovery and purification fold of lysozyme obtained from fresh chicken egg whites added with 1. 0% ascorbic acid were 78% and 2.4, respectively. Storage tests showed that the obtained lyophilized lysozyme powder after dialysis was stable when refrigerated at 4 degrees C for 3 months. PMID- 10691611 TI - Temperature and exposure time during ethylene conditioning affect ripening of Bartlett pears. AB - Freshly harvested early- and mid-season Bartlett pears (Pyrus communis) were treated with ethylene (air plus 10 Pa C(2)H(4)) or air at 5, 10, and 20 degrees C for 24 and 48 h (experiment 1) and at 5 and 10 degrees C for 48, 72, and 96 h and at 20 degrees C for 24 h (experiment 2). Following C(2)H(4) or air treatment at different temperatures and durations, pears were transferred to 20 degrees C in air for ripening. Bartlett pears were evaluated for firmness, color, respiration, C(2)H(4) production, and activities of 1-aminocyclopropane-1-carboxylic acid synthase (ACC-S) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACC-O). Ethylene action was temperature dependent. The duration of C(2)H(4) conditioning needed to fully induce ripening was longer at lower temperatures: 72 h at 5 degrees C, 48 h at 10 degrees C, and 24 h at 20 degrees C. Cold storage in air for as little as 3-4 days at 5 or 10 degrees C appeared to hasten subsequent ripening, but to a lesser extent than pears kept for 2 weeks at -1 degrees C in air. Despite a significant increase in ACC-S activity in pears treated with C(2)H(4) at 5 degrees C, there was not a simultaneous increase in ACC-O activity, resulting in low C(2)H(4) production that was insufficient to generate the threshold endogenous levels of C(2)H(4) required for ripening. Contrary to previous findings with pears, these data indicate that ACC-O could be a rate limiting step in C(2)H(4) biosynthesis. PMID- 10691612 TI - Purification and characterization of a stable cysteine protease ervatamin B, with two disulfide bridges, from the latex of Ervatamia coronaria. AB - Latex of the medicinal plant Ervatamia coronaria was found to contain at least three cysteine proteases with high proteolytic activity, called ervatamins. One of these proteases, named ervatamin B, has been purified to homogeneity using ion exchange chromatography and crystallization. The molecular mass of the enzyme was estimated to be 26 000 Da by SDS-PAGE and gel filtration. The extinction coefficient (epsilon(1%)(280 nm)) of the enzyme was 20.5 with 7 tryptophan and 10 tyrosine residues per molecule. The enzyme hydrolyzed denatured natural substrates such as casein, azoalbumin, and azocasein with a high specific activity. In addition, it showed amidolytic activity toward N-succinyl-alanine alanine-alanine-p-nitroanilide with an apparent K(m) and K(cat) of 6.6 +/- 0.5 mM and 1.87 x 10(2) s(-)(1), respectively. The pH optima was 6.0-6.5 with azocasein as substrate and 7.0-7.5 with azoalbumin as substrate. The temperature optimum was around 50-55 degrees C. The enzyme was basic with an isoelectric point of 9.35 and had no carbohydrate content. Both the proteolytic and amidolytic activity of the enzyme was strongly inhibited by thiol-specific inhibitors. Interestingly, the enzyme had only two disulfide bridges versus three as in most plant cysteine proteases of the papain superfamily. The enzyme was relatively stable toward pH, denaturants, temperature, and organic solvents. Polyclonal antibodies raised against the pure enzyme gave a single precipitin line in Ouchterlony's double immunodiffusion and typical color in ELISA. Other related proteases do not cross-react with the antisera to ervatamin B showing that the enzyme is immunologically distinct. The N-terminal sequence showed conserved amino acid residues and considerable similarity to typical plant cysteine proteases. PMID- 10691613 TI - Antioxidative and anti-glycation activity of garcinol from Garcinia indica fruit rind. AB - Garcinol, a polyisoprenylated benzophenone derivative, was purified from Garcinia indica fruit rind, and its antioxidative activity, chelating activity, free radical scavenging activity, and anti-glycation activity were studied. Garcinol exhibited moderate antioxidative activity in the micellar linoleic acid peroxidation system and also exhibited chelating activity at almost the same level as citrate. It also showed nearly 3 times greater DPPH (1, 1-diphenyl-2 picrylhydrazyl) free radical scavenging activity than DL-alpha-tocopherol by weight in aqueous ethanol solution. In a phenazine methosulfate/NADH-nitroblue tetrazolium system, garcinol exhibited superoxide anion scavenging activity and suppressed protein glycation in a bovine serum albumin/fructose system. Thus, garcinol might be beneficial as a potent antioxidant and a glycation inhibitor under specified conditions. PMID- 10691614 TI - Protein insolubilization and thiol oxidation in sulfite-treated wheat gluten films during aging at various temperatures and relative humidities. AB - Gluten films were prepared by casting an acidic and ethanolic solution of gluten previously treated with sodium sulfite. The effects of sulfitolysis on proteins were investigated by SE-HPLC and thiol/disulfide content measurements. During sulfitolysis, insoluble glutenin macropolymer was converted into its constitutive subunits. About 10% of gluten disulfide bonds were cleaved, of which three fourths originated from interchain disulfide bonds. Oxidation of thiol groups released during sulfitolysis was followed for various temperatures (T) and relative humidities. Oxidation was shown to be a second-order rate process occurring below the glass transition temperature (T(g)) and related to T - T(g). Thiol oxidation ensured the formation of interchain bonds between specific classes of gluten proteins according to an ordered process. Intrachain bonds were also formed and through thiol/disulfide-exchange reactions were finally converted to interchain bonds. Thus, fully oxidized gluten films had more insoluble glutenin macropolymers than native gluten. PMID- 10691615 TI - Development of oxidized odor and volatile aldehydes in fermented cucumber tissue exposed to oxygen. AB - Changes in volatile compounds in fermented cucumber tissue during exposure to oxygen were investigated by purge and trap sampling, followed by GC-MS. Hexanal and a series of trans unsaturated aldehydes, (E)-2-pentenal, (E)-2-hexenal, (E)-2 heptenal, and (E)-2-octenal, increased in fermented cucumber slurries exposed to oxygen. Sensory evaluation of oxidized odor was correlated with the increase in aldehyde concentrations. Other identified volatile components present after fermentation did not show major changes during exposure to oxygen. There was no decrease in the formation of aldehydes in fermented cucumber samples that were heated to inactivate enzymes before exposure to oxygen. These results indicated that the formation of aldehydes in oxygen was due to nonenzymatic reactions. PMID- 10691616 TI - Orthokinetic flocculation of caseinate-stabilized emulsions: influence of calcium concentration, shear rate, and protein content. AB - Calcium-induced flocculation of caseinate-stabilized soybean oil-in-water emulsions in conditions of Couette flow was studied. A concentrated emulsion (20% oil, 0.5-2.0% sodium caseinate in 20 mM imidazole, pH 7) was diluted 20 times in buffer containing concentrations of CaCl(2) between 9 and 17 mM and sheared at rates between 335 and 1340 s(-)(1). The average particle size (d(43)), measured by integrated light scattering, increased in a sigmoidal manner with shearing time. An increased shear rate resulted in an increased flocculation rate, because of the increased number of collisions between particles, but a decreased value of the maximum d(43), because higher shear rates increasingly disrupted the flocs. The flocculation rate was increased by increasing the calcium concentration, indicating an increased collision efficiency. The orthokinetic stability of the emulsions was increased with increased protein content, and it is postulated that the increased surface coverage and hydrodynamic thickness of the adsorbed protein layer increased steric repulsion between droplets, so that higher calcium concentrations were necessary to induce sufficient conformational change of the proteins to allow flocculation. At high caseinate concentrations, calcium may also induce precipitation of unadsorbed caseins from the serum to the oil-water interface, thereby increasing steric repulsion and hence increasing orthokinetic stability. PMID- 10691617 TI - Protein binding in deactivation of ferrylmyoglobin by chlorogenate and ascorbate. AB - Kinetics of reduction of iron(IV) in ferrylmyoglobin by chlorogenate in neutral or moderately acidic aqueous solutions (0.16 M NaCl) to yield metmyoglobin was studied using stopped flow absorption spectroscopy. The reaction occurs by direct bimolecular electron transfer with (2.7 +/- 0.3) x 10(3) M(-)(1).s(-)(1) at 25.0 degrees C (DeltaH( )(#) = 59 +/- 6 kJ.mol(-)(1), DeltaS(#) = 15 +/- 22 J. mol( )(1).K(-)(1)) for protonated ferrylmyoglobin (pK(a) = 4.95) and with 216 +/- 50 M(-)(1).s(-)(1) (DeltaH( )(#) = 73 +/- 8 kJ. mol(-)(1), DeltaS( )(#) = 41 +/- 30 J.mol(-)(1).K(-)(1)) for nonprotonated ferrylmyoglobin in parallel with reduction of a chlorogenate/ferrylmyoglobin complex by a second chlorogenate molecule with (8.6 +/- 1.1) x 10(2) M(-)(1).s(-)(1) (DeltaH( )(#) = 74 +/- 8 kJ.mol(-)(1), DeltaS( )(#) = 59 +/- 28 J.mol(-)(1).K(-)(1)) for protonated ferrylmyoglobin and with 61 +/- 9 M(-)(1).s(-)(1) (DeltaH( )(#) = 82 +/- 12 kJ.mol(-)(1), DeltaS( )(#) = 63 +/- 41 J. mol(-)(1).K(-)(1)) for nonprotonated ferrylmyoglobin. Previously published data on ascorbate reduction of ferrylmyoglobin are reevaluated according to a similar mechanism. For both protonated and nonprotonated ferrylmyoglobin the binding constant of chlorogenate is approximately 300 M(-)(1), and the modulation of ferrylmyoglobin as an oxidant by chlorogenate (or ascorbate) leads to a novel antioxidant interaction for reduction of ferrylmyoglobin by ascorbate in mixtures with chlorogenate. PMID- 10691618 TI - Iron-accelerated cumene hydroperoxide decomposition in hexadecane and trilaurin emulsions. AB - Free radicals arising from lipid peroxides accelerate the oxidative deterioration of foods. To elucidate how lipid peroxides impact oxidative reactions in food emulsions, the stability of cumene hydroperoxide was studied in hexadecane or trilaurin emulsions stabilized by anionic (sodium dodecyl sulfate; SDS), nonionic (Tween 20), and cationic (dodecyltrimethylammonium bromide; DTAB) surfactants. Fe(2+) rapidly (within 10 min) decomposed between 10 and 31% of the cumene hydroperoxide in Tween 20- and DTAB-stabilized emulsions at pH 3.0 and 7.0 and in the SDS-stabilized emulsion at pH 7.0 with no further decomposition of peroxides occurring for up to 3 h. In SDS-stabilized emulsions at pH 3.0, Fe(2+) decreased peroxides by 90% after 3 h. Decomposition of peroxides in the absence of added iron and by Fe(3+) was observed only in SDS-stabilized emulsions at pH 3.0. These results suggest that peroxide decomposition by iron redox cycling occurs when iron emulsion droplet interactions are high. PMID- 10691619 TI - Relationship among antioxidant activity, vasodilation capacity, and phenolic content of red wines. AB - The relationship among antioxidant activity, based on the electron-spin resonance determination of the reduction of Fremy's radical, vasodilation activity, and phenolic content was investigated in 16 red wines. The wines were selected to provide a range of origins, grape varieties, and vinification methods. Sensitive and selective HPLC methods were used for the analysis of the major phenolics in red wine: free and conjugated myricetin, quercetin, kaempferol, and isorhamnetin; (+)-catechin, (-)-epicatechin, gallic acid, p-coumaric acid, caffeic acid, caftaric acid, trans-resveratrol, cis-resveratrol, and trans-resveratrol glucoside. Total anthocyanins were measured using a colorimetric assay. The total phenolic content of the wines was determined according to the Folin-Ciocalteu colorimetric assay and also by the cumulative measurements obtained by HPLC. The 16 wines exhibited a wide range in the values of all parameters investigated. However, the total phenol contents, measured both by HPLC and colorimetrically, correlated very strongly with the antioxidant activity and vasodilation activity. In addition, the antioxidant activity was associated with gallic acid, total resveratrol, and total catechin. In contrast, only the total anthocyanins were correlated with vasodilation activity. The results demonstrate that the different phenolic profiles of wines can produce varying antioxidant and vasodilatant activities, which opens up the possibility that some red wines may provide enhanced health benefits for the consumer. PMID- 10691620 TI - Unsaponifiable lipid constituents of some underutilized tropical seed oils. AB - Sterols, triterpene alcohols, and hydrocarbons present in the unsaponifiable fraction of some underutilized tropical seed oils have been examined. The seeds include Telfairia occidentalis (TLO), Andenopus breviflorus (ADB), Cucumeropsis edulis (CME), Antiaris africana (ATF), and Monodora tenuifolia (MNT). The oil content of the seeds was high (34.7-68.8%), whereas triacylglycerols comprised the dominant lipid group in the oils (65.4-73.9%). The percentage of unsaponifiables ranged from 1.1 to 7.9%. Ten sterols were identified in the fractions. In the Cucurbitaceae oils (TLO, CME, and ADB), Delta(7)-sterols constituted the dominant sterols. These include 24-ethylcholesta-7,22E,25-trienol (7), 24-ethylcholesta-7,25-dienol (9), 24Z-ethylidenecholes-7-enol (10), and 24 ethylcholesta-7, 24-dienol (11). However Delta(5)-sterols (1-5) occurred at the highest concentration in the other two samples (ATF and MNT). Fifteeen triterpene alcohols were detected in the fractions. Olean-12-enol (16), isomultiflorenol (8), and lupeol (23) were the dominant alcohols in the Cucurbitaceae family, whereas alpha-amyrin (urs-12-enol) (20) was the dominant triterpene alcohol in ATF and MNT. A mixture of C(18)-C(34) n-alkanes, squalene, and some monoterpenes was detected in the hydrocarbon fraction. PMID- 10691621 TI - Isolation and structural elucidation of two new glycosides from sage (Salvia officinalis L.). AB - Six compounds, 1-O-(2,3, 4-trihydroxy-3-methyl)butyl-6-O-feruloyl-beta-D glucopyranoside, ethyl beta-D-glucopyranosyl tuberonate, p-hydroxybenzoic acid, ( )-hydroxyjasmonic acid, caffeic acid, and 4-hydroxyacetophenone 4-O-[5-O-(3, 5 dimethoxy-4-hydroxybenzoyl)-beta-D-apiofrunosyl]-(1-->2)-beta-D- glu copyranoside, were isolated from the n-butanol-soluble fraction of sage leaf extracts. Their structures were determined by spectral methods (MS, NMR, and 2D NMR), and their antioxidant activities were measured. Among them, two new glycosides were elucidated. All of these compounds showed DPPH free radical scavenging activity at the concentration of 30 mM, and caffeic acid was the most active compound. PMID- 10691622 TI - Use of two-dimensional electrophoresis to evaluate proteolysis in salmon (Salmo salar) muscle as affected by a lactic fermentation. AB - Two-dimensional electrophoresis was used to study proteolysis in salmon fillets inoculated or not with the starter culture Lactobacillus sake LAD. Protein fragments appeared increasingly with time in both samples, indicating that the main quantitative changes were due to endogenous enzymes. In the most acidic zone (pI = 4-6. 20) particularly, proteolysis was overall independent from processing. In contrast, fermentation had a significant effect in the pH range 6.20-8.35, suggesting a specificity of the bacterial proteases of L. sake toward alkaline to slightly acidic proteins. Furthermore, fragments surrounding tropomyosin (apparent pI = 4.70) appeared in fermented samples, indicating that the protein may be a suitable substrate for the metabolism of L. sake LAD. PMID- 10691623 TI - Distribution and characterization of enzymes causing starch degradation in rice (Oryza sativa cv. koshihikari). AB - The thermal dependency and stability of enzymes producing reducing sugar (RS) were examined in bran, the exterior 13% part (outer endosperm), and the remaining inner endosperm of rice grains. RS-producing enzymes in the inner endosperm showed a higher optimum temperature than those in other parts of the rice grain. Diethylaminoethyl-Sephacel chromatography of crude extracts revealed two peaks of RS-producing activity with different optimum temperatures (60 and 37 degrees C) in all three parts. alpha-Glucosidase (EC 3.2.1.20) and alpha-amylase (EC 3.2.1.1) isoform G were thought to be major components of the RS-producing activities with high and low optimum temperatures, respectively. The peak with a high optimum temperature was a more abundant component in the inner endosperm, compared with other parts of the rice grain. Thus, different parts of rice were found to have distinct enzyme sets having different thermal dependency and to be involved in starch degradation to various sugars. PMID- 10691624 TI - Isolation and identification of stilbenes in two varieties of Polygonum cuspidatum. AB - The roots of two varieties of Polygonum cuspidatum (Hu Zhang and Mexican Bamboo) were analyzed for resveratrol and analogues. The roots of each variety were dried and ground into a powder. The powdered roots were then extracted with methanol and ethyl acetate. The ethyl acetate fraction of the Mexican Bamboo was then subjected to fractionation and purification using silica gel column chromatography and semipreparative HPLC. In addition to resveratrol (3,5,4' trihydroxystilbene), three stilbene glucosides were identified by (1)H NMR, (13)C NMR, and MS. The stilbene glucosides were shown to be a piceatannol glucoside (3,5,3', 4'-tetrahydroxystilbene 4'-O-beta-D-glucopyranoside), resveratroloside (3,5,4'-trihydroxystilbene 4'-O-beta-D-glucopyranoside), and piceid (3,5,4' trihydroxystilbene 3-O-beta-D-glucopyranoside). The levels of the piceatannol glucoside and piceid were twice as high in the Mexican Bamboo as compared to the Hu Zhang. PMID- 10691625 TI - Purification and partial characterization of a second cysteine proteinase inhibitor from ungerminated barley (Hordeum vulgare L.). AB - It was previously shown that ungerminated barley contains inhibitors that suppress the activities of green malt cysteine proteinases. This paper reports the purification and partial characterization of a second barley cysteine endoproteinase inhibitor, a protein called lipid transfer protein 2 (LTP2). The chromatographically purified inhibitor had a molecular mass of 7112. The amino acid composition and sequence data of the purified inhibitor indicated that it was a protein whose gene, but not the protein itself, was isolated earlier from barley aleurone tissue. The purified protein inhibited the activities of electrophoretically separated green malt cysteine proteinases but not the activities of the serine- or metalloproteinases. The purified LTP2 inhibited the same proteases as the LTP1 that was characterized previously but was present in the mature seed in much smaller amounts. Neither LTP1 nor LTP2 has been proven to transport lipids in vivo, and it seems possible that both serve to keep cysteine endoproteinases that are synthesized during barley seed development inactive until the plant needs them. The small amount of LTP2 in the seed made it impossible to determine whether it, like LTP1, is involved in beer foam formation. Because of its proteinase-inhibiting ability and its resistance to heat inactivation, some of the LTP2 may persist in beer. PMID- 10691626 TI - Lipophilization of lysozyme by short and middle chain fatty acids. AB - Hen egg white lysozyme was lipophilized with short and middle chain saturated fatty acids (caproic, capric, or myristic acid). The yield, bactericidal properties, and structural properties of lipophilized lysozymes were investigated. The yield of lipophilization of lysozyme greatly increased with the decrease in the chain length of fatty acid. Lipophilization broadened the bactericidal action of lysozyme to Gram-negative bacteria with little loss of enzymatic activity. The bactericidal activity increased in proportion to the number of bound short chain fatty acids. The thermal stability of lipophilized lysozyme decreased in proportion to the chain length and number of bound fatty acids. PMID- 10691627 TI - Isolation of homogeneous fractions from wheat water-soluble arabinoxylans. Influence of the structure on their macromolecular characteristics. AB - Water-soluble arabinoxylans from wheat flour were purified and fractionated by graded ethanol precipitation. Six fractions were obtained at 20% (F20), 30% (F30), 40% (F40), 50% (F50), 60% (F60), and 70% (F70) saturation with ethanol. Neutral sugars and (1)H NMR analyses revealed differences in structural characteristics. The Ara/Xyl ratio and the amount of Xylp residues disubstituted increased with ethanol concentration. Ferulic acid content was higher in fractions precipitated at low ethanol percentage. Fractions were refractionated by SEC, leading to 46 subfractions with low polydispersity index. Substitution degree was apparently linearly related to the amount of disubstituted Xylp. Macromolecular characteristics (M(w), [eta], R(G), q, nu) determined by multiangle laser light scattering and viscosimetry were similar among all fractions. A rather flexible conformation was determined for the arabinoxylans, in conflict with the admitted rodlike conformation. The substitution degree had no influence on the conformation or on the rigidity of the polymers. Evidence for the presence of ferulic acid dimers in the water-soluble arabinoxylans is provided, which probably explains the unexpected conformation and macromolecular characteristics. PMID- 10691628 TI - Kinetics and mechanism of the primary steps of degradation of carotenoids by acid in homogeneous solution. AB - The kinetics of reaction between trifluoroacetic acid as an acid of medium strength and the carotenoids beta-carotene, zeaxanthin, canthaxanthin, and astaxanthin has been examined in detail including the effects of dioxygen, acid concentration, and carotenoid structure. Reaction between acid and carotenoid leads to species absorbing in the red and near-infrared (NIR) spectral regions, intermediates that subsequently disappear. ESR experiments clearly show that these species are not carotenoid radicals, although their NIR absorption is similar to the absorption of carotenoid radical cations. Under most reaction conditions, the disappearance of carotenoids follows pseudo-zero-order kinetics, whereas the reaction order is >1 with respect to acid, and the long-lived (hours) intermediates are suggested to be mono- (700 nm) and diprotonated carotenoid ( approximately 950 nm). Acid induces cis/trans-isomerization via the protonated intermediates, which also decay to nonradical species with shorter conjugated systems-most probably carotenoid esters. Slow protonization of the methine carbon is the primary step in the degradation, but dioxygen increases the rate as a result of formation of a charge-transfer complex with the carotenoids as indicated by a red-shift of the NIR absorption bands. Carotenoids with carbonyl groups (astaxanthin and canthaxanthin) have slower rates of degradation than beta carotene and zeaxanthin, indicating preferential nondegradative protonation of the carbonyl groups. PMID- 10691629 TI - HPLC detection of soluble carbohydrates involved in mannitol and trehalose metabolism in the edible mushroom Agaricus bisporus. AB - A convenient and sensitive method was developed to separate and detect various types of carbohydrates (polyols, mono- and disaccharides, and phosphorylated sugars) simultaneously using high-performance liquid chromatography (HPLC). The method consists of a chromatographic separation on a CarboPac PA1 anion-exchange analytical column followed by pulsed amperometric detection. In a single run (43 min) 13 carbohydrates were readily resolved. Calibration plots were linear over the ranges of 5-25 microM to 1. 0-1.5 mM. The reliable and fast analysis technique, avoiding derivatization steps and long run times, was used to determine the levels of carbohydrates involved in mannitol and trehalose metabolism in the edible mushroom Agaricus bisporus. Moreover, the method was used to study the trehalose phosphorylase reaction. PMID- 10691630 TI - Effect of calcium on the oxidation of linoleic acid by potato (Solanum tuberosum var. Desiree) tuber 5-lipoxygenase. AB - When the effect of calcium on the oxidation of linoleic acid by potato tuber 5 lipoxygenase (LOX) was investigated, it was seen to promote the enzyme's activity at pH values higher than the optimum pH of 6.3, resulting in an enzyme activation at alkaline pH. Kinetic analysis of calcium activation at different pH values revealed that the cation abolished the inhibition by high substrate concentration, which occurs in the absence of Ca(2+), thus leading to activation at high substrate concentration. Studies were conducted to investigate the influence of Ca(2+) on the physicochemical nature of the substrate and its effect on the LOX activity expression. It was concluded that the aggregation mode rather than the aggregation state of linoleic acid is responsible for potato 5-LOX changes. PMID- 10691631 TI - Enzymatic assay for the determination of olive oil polyphenol content: assay conditions and validation of the method. AB - A new spectrophotometric assay for the determination of the polyphenolic content of olive oil is presented. It is a substrate-recycling assay for phenolic compounds that employs tyrosinase in the presence of excess NADH. The reaction of various phenols with the enzyme produces an o-quinone, which is detected by recycling between reactions with the enzyme and NADH. The method offers some advantages over the classical methods employed to determine the polyphenolic content of olive oil, that is, ease and reproducibility of the analysis, highly increased sensitivity, and selectivity toward phenolic compounds. The amount of total polyphenols was determined in virgin olive oils both with the Folin Ciocalteu reagent and with the proposed enzymatic method. The results suggest a better estimation of the polyphenol content, as compared with the colorimetric method. This has to be attributed to the different reactivities of the two methods toward phenols and catechols. Finally, the enzymatic method demonstrates that there is a linear relationship between the olive oil phenolic content and the antioxidative capacity of oil extracts. PMID- 10691632 TI - Effect of electron microscopy fixation pH on the ultrastructure of soybean protein bodies. AB - Cotyledon tissues from mature soybeans were systematically prepared for transmission electron microscopy employing fixation buffer pH's of 7.2, 6.4, and 5.6. Tissue fixed at pH 7.2 showed few membrane-bound internal protein body structures. Portions of the same tissue fixed at pH 6.4 revealed numerous membrane-bound crystalloid structures and stacks of membranous sacks. Tissue prepared at pH 5.6 also contained numerous membrane-bound crystalloid structures and examples of membrane-bound globoid structures. This is the first investigation to document the effect of fixation pH on the subcellular structure of soybean protein bodies and the first to show membrane-bound crystalloid or globoid structures in soybeans. PMID- 10691633 TI - Synthesis of haptens and conjugates for ELISAs of phytoestrogens. Development of the immunological tests. AB - Seven carboxylic acid haptens of isoflavonoids were synthesized, with the spacer arm on the oxygen atom at the C7 position for one series, with formononetin, daidzein, equol, biochanin A, and genistein, and at the C8 position for a second series, with only formononetin and daidzein. The different haptens were coupled to bovine serum albumin (BSA) and to swine thyroglobulin (Thyr). Polyclonal antibodies were generated against the BSA conjugates. Enzyme-linked immunosorbent assays (ELISAs) were developed based on competition between free phytoestrogens and the Thyr-hapten conjugates for specific antibodies. IC(50) values of the standard curves ranged between 0.8 and 20 ng/mL that is, 0.3 and 9.2 pmol/well. The antibodies obtained should be useful for assays in vegetable matter as well as in biological fluids after a separation step. These ELISAs should be valuable also in the food industry to control phytoestrogen concentrations prior to and after processing. PMID- 10691634 TI - Simultaneous stopped-flow determination of butylated hydroxyanisole and propyl gallate using a T-format luminescence spectrometer. AB - A simple and fast luminescent method is used for the first time to resolve a mixture of two synthetic antioxidants, propyl gallate (PG) and butylated hydroxyanisole (BHA), by the joint use of the stopped-flow mixing technique and a T-format luminescence spectrometer. The determination of these compounds involves two different and independent reactions. On the one hand, PG determination is based on an energy transfer process that involves the formation of a lanthanide chelate with terbium in the presence of Triton X-100 and tri-n-octylphosphine oxide. On the other hand, BHA is determined using a reaction between the oxidized form of Nile Blue and the antioxidant. Both systems are excited at the same excitation wavelength (310 nm), and the emission wavelengths are 545 and 665 nm for PG and BHA, respectively. The absence of overlap in the emission spectra makes it possible to measure separately the analytes in each channel of the instrument. Initial rate and equilibrium signal are used as analytical parameters and measured in 0.1 and 1 s for PG and BHA, respectively. Calibration graphs are linear over the range 0.09-3.5 microg mL(-)(1) for PG and 0.3-15 microg mL(-)(1) for BHA. The relative standard deviations of both systems are close to 2%. The proposed method is applied to the determination of these two antioxidants in several commercial food samples with recoveries ranging between 94.8 and 102.9% for PG and between 94.1 and 102.1% for BHA. PMID- 10691635 TI - Acetonitrile as a buffer additive for free zone capillary electrophoresis separation and characterization of maize (Zeamays L. ) and sorghum (Sorghum bicolor L. Moench) storage proteins. AB - An improved method for separating and characterizing maize (Zea mays L.) and sorghum (Sorghum bicolor L. Moench) storage proteins by free zone capillary electrophoresis (FZCE) was developed. Previous electrophoretic methods for analyzing these proteins required high concentrations of urea to maintain protein solubility during separation. To overcome disadvantages of urea, we developed a FZCE method that mimicked reversed-phase high-performance liquid chromatography (RP-HPLC) in that it used high levels of acetonitrile (ACN) at low pH. The optimized FZCE buffer system consisted of 80 mM phosphate-glycine buffer, nominal pH 2.5, containing 60% ACN and a cellulose derivative to dynamically coat capillary walls. Resolution was similar to or higher than that previously achieved by FZCE buffers utilizing 8 M urea as a buffer additive. ACN concentrations of at least 50% were necessary to achieve acceptable separations; this ACN concentration is approximately that necessary to extract these storage proteins. ACN was equally effective as traditional ethanol solvents and 8 M urea for solubilizing maize and sorghum proteins. The ACN-based FZCE buffer system gave high repeatability (<0.3% relative standard deviation, measured over 15 consecutive injections) for migration time. Subclasses of maize and sorghum storage proteins were identified, and genotypes of each cereal were successfully differentiated using ACN-containing buffers. This FZCE method may be applicable for the analysis of other hydrophobic proteins without the use of urea. PMID- 10691636 TI - Comparison of protein surface hydrophobicity measured at various pH values using three different fluorescent probes. AB - The influence of type of fluorescent probe on the surface hydrophobicity values determined for three native and heated proteins was assessed using uncharged [6 propionyl-2-(N, N-dimethylamino)naphthalene or PRODAN] versus anionic aliphatic (cis-parinaric acid or CPA) and aromatic (1-anilinonaphthalene-8-sulfonic acid or ANS) probes. Surface hydrophobicities of whey protein isolate, beta lactoglobulin, and bovine serum albumin under heated (80 degrees C for 30 min) and unheated conditions and at varying pH values (3.0, 5.0, 7.0, and 9. 0) were measured using ANS, CPA, and PRODAN. ANS and CPA yielded opposing results for the effects of pH and heating on protein hydrophobicity. Hydrophobicity was lower at pH 3.0 than at other pH values for all proteins measured by PRODAN, whereas the values measured by ANS and CPA at pH 3.0 were quite high compared to those at other pH values, suggesting the influence of electrostatic interactions on anionic probe-protein binding. These results suggest that the presence or absence of a permanent charge as well as the aromatic and aliphatic nature of fluorescent probes can affect protein hydrophobicity values measured under various pH conditions. PMID- 10691637 TI - Functional scFv antibody sequences against the organophosphorus pesticide chlorpyrifos. AB - Two functional single-chain Fv (scFv) antibodies that recognize specifically the widely used organophosphorus pesticide chlorpyrifos-ethyl were derived from two murine hybridoma cell lines. It is shown that the functional scFvs could be isolated without any rounds of selection, with a success rate dependent on the efficiency of amplification of the functional light chain gene family by the specific primers. Besides four new functional immunoglobulin variable gene sequences, the isolation of a new pseudogene is reported. PMID- 10691638 TI - Separation and purification of anthocyanins by high-speed countercurrent chromatography and screening for antioxidant activity. AB - The all-liquid chromatographic technique of high-speed countercurrent chromatography (HSCCC) has been applied for separations of anthocyanins. The biphasic mixture of tert-butyl methyl ether/n-butanol/acetonitrile/water (2:2:1:5) acidified with trifluoroacetic acid was found to be a suitable solvent system for anthocyanin separation. In some cases, enrichment of the pigments on Amberlite XAD-7 resin prior to HSCCC has been carried out. The anthocyanin mixtures from red cabbage, black currant, black chokeberry, and roselle were successfully fractionated using HSCCC. Peak purity control was done by nuclear magnetic resonance spectroscopy as well as electrospray ionization ion trap multiple mass spectrometry. Finally, antioxidant activity of the purified pigments was determined using the Trolox equivalent antioxidant capacity test. PMID- 10691639 TI - Ultrafast capillary electrophoretic analysis of cereal storage proteins and its applications to protein characterization and cultivar differentiation. AB - Free zone capillary electrophoresis conditions have been improved to allow rapid (2-8 min) separations of grain proteins from several cereals (wheat, oats, rice, barley, and rye) with high resolution and reproducibility. This new method utilized the isoelectric compound iminodiacetic acid (IDA) in conjunction with 20% acetonitrile and 0.05% hydroxypropylmethylcellulose. Cultivars of all cereals tested could be differentiated in 3 min, including wheat, using either prolamin or glutelin protein patterns. Resolution was similar to or higher than that of separations in other acidic buffers. Migration time repeatability was excellent with run-to-run variability <1% RSD, day-to-day <1.4% RSD, and capillary-to capillary <3.3% RSD. Because larger inner diameter capillaries (50 microm) could be used with this buffer, sensitivity was improved and capillary rinse times could be reduced when compared to smaller capillaries (25 microm i.d.). This also served to reduce total separation time so that the majority of cereal storage protein from several types of cereals could be analyzed with total analysis times of 2-8 min with extremely high resolution and repeatability. This method would allow unattended, high-throughput ( approximately 180-400 samples/24 h) analysis of cereal proteins without the generation of much organic solvent waste as well as automated data analysis and storage. PMID- 10691640 TI - LC-ESI-MS study of the flavonoid glycoside malonates of red clover (Trifolium pratense). AB - High-performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was applied to the analysis of the flavonoids and their glycoside malonates of the flowers and leaves of red clover (Trifolium pratense). Through LC-MS comparative studies on the plant extracts and their malonate-free extracts, approximately 20 flavonoid glycoside malonates were detected in the flower extract. Eight were identified as genistin 6' '-O-malonate (39), formononetin 7-O beta-D-glucoside 6' '-O-malonate (40), biochanin A 7-O-beta-D-glucoside 6' '-O malonate (41), trifoside 6' '-O-malonate (42), irilone 4'-O-beta-D-glucoside 6' ' O-malonate (43), pratensein 7-O-beta-D-glucoside 6' '-O-malonate (44), isoquercitrin 6' '-O-malonate (45), and 3-methylquercetin 7-O-beta-D-glucoside 6' '-O-malonate (46). About 15 other flavonoids and clovamides were proved to be present in this extract. The study also found that the flowers contained flavones as the major flavonoids, whereas the leaves had isoflavones as the major flavonoids. This is the first detection of the six malonates (39 and 42-46) in the extracts of red clover, and among them, 42, 43, and 46 are new compounds. PMID- 10691641 TI - Determination of spinosad and its metabolites in citrus crops and orange processed commodities by HPLC with UV detection. AB - Spinosad is an insect control agent that is derived from a naturally occurring organism and is effective on a wide variety of crops, including citrus crops. A method is described for the determination of spinosad and its metabolites in citrus crops and orange processed commodities. The method determines residues of the active ingredients (spinosyns A and D) and three minor metabolites (spinosyn B, spinosyn K, and N-demethylspinosyn D). For dried orange pulp and orange oil, the method has a limit of quantitation (LOQ) of 0.02 microg/g and a limit of detection (LOD) of 0.006 microg/g. For all other sample matrices (whole fruit, edible fruit, juice, and peel), the method has an LOQ of 0.01 microg/g and an LOD of 0.003 microg/g. The analytes are extracted from the various sample types using appropriate solvents, and the extracts are purified by liquid-liquid partitioning and/or solid-phase extraction. All five analytes are determined simultaneously in the purified extracts by reversed-phase high-performance liquid chromatography with ultraviolet detection at 250 nm. PMID- 10691642 TI - Isolation of novel glucosides related to gingerdiol from ginger and their antioxidative activities. AB - Two novel glucosides of 6-gingerdiol were isolated from fresh ginger (Zingiber officinale Roscoe). Their structures were determined as 1-(4-O-beta-D glucopyranosyl-3-methoxyphenyl)-3,5-dihydroxydecane (1) and 5-O-beta-D glucopyranosyl-3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)deca ne (2) by HRFAB-MS and NMR analyses, and the absolute configurations of both aglycons were identified as (3S,5S) by a comparison with synthetic compounds. After incubating these glucosides with acetone powder prepared from fresh ginger, they were confirmed to have been hydrolyzed to 6-gingerdiol by HPLC. This result suggests that these glucosides are the precursors or intermediates of 6-gingerdiol. To recognize their function, their antioxidative activities were investigated and compared to that of their aglycon, 6-gingerdiol, by a linoleic acid model system and by their DPPH radical-scavenging ability. Although 1 did not indicate any activity, 2 had as strong activity as the aglycon in both measurements. PMID- 10691643 TI - First report of the characterization of the threatened plant species Amaranthus pumilus (Seabeach amaranth). AB - This paper reports the first ever investigation of the chemical components/composition of the seeds of Amaranthus pumilus (a threatened amaranth species) and compares the results to those of the more commonly cultivated Amaranthus hypochondriacus (Plainsman). This study clearly revealed that much genetic diversity exists between these species, indicating that potential breeding possibilities for the improvement of more commonly cultivated amaranth lines do exist. A. pumilus offers a much larger and more desirable seed size and weight (2-3-fold higher), permitting greater biomass production, in addition to lower levels (half) of free carbohydrate for improved value/performance in diabetic-type diets. In addition to the higher edible oil content found in A. pumilus, its lower saturated/unsaturated ratio (one of the lowest thus far measured) makes it potentially a better source of nutritional oil. In addition to the 2-fold-higher quantity of vitamin E found in A. pumilus, the higher levels of squalene also found may one day serve as a renewable crop source of this compound and may diminish the world's dependence upon marine animals. Considering the imminent danger posed to the survival of the seabeach amaranth in its native environment, it is hoped this study will raise public awareness of the importance of this species and thereby protect it from reaching extinction. PMID- 10691644 TI - Composition of suberin extracted upon gradual alkaline methanolysis of Quercus suber L. cork. AB - The monomeric composition of suberin extracts obtained by gradual alkaline methanolysis of Quercus suber cork was determined by gas chromatography-mass spectrometry (GC-MS). Results show that 1-alkanols and alkanoic and alpha,omega alkanedioic acids are preferentially removed upon mild alkaline conditions, whereas mid-chain-modified omega-hydroxyalkanoic acids are preferentially removed under stronger alkaline conditions. Saturated omega-hydroxyalkanoic acids are found to be abundant in all suberin extracts. These results are consistent with two distinct suberin fractions with different locations in cork cell walls and/or esterification degrees. It is proposed that these fractions correlate with the two main suberin peaks in the solid state (13)C NMR spectra of cork and suberin extracts. Quantitative GC-MS analysis showed that suberin monomers comprise approximately 30% (w/w) of the suberin extracts, the remaining comprising nonvolatile structures with high M(n) values, as measured by vapor pressure osmometry. The presence of a large fraction of high molecular weight aliphatic structures in suberin extracts is supported by the corresponding NMR spectra. PMID- 10691645 TI - N-Nitrosation of myosmine yields HPB (4-hydroxy-1-(3-pyridyl)-1-butanone) and NNN (N-nitrosonornicotine). AB - N-Nitrosonornicotine (NNN) is formed by synthetic or biological N-nitrosation of the tobacco alkaloid nornicotine. Following metabolic activation of NNN, DNA and protein adducts are formed releasing 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB), an actual biomarker to differentiate between tobacco smokers and passive smokers. NNN and HPB can be prepared in a new one-step reaction by N-nitrosation of the nicotinoid myosmine which has been found not only in tobacco but also in nut products. The reaction was tested also in human gastric juice. The formation rate of NNN and HPB depends on the pH value in the reaction solutions. This is important under the aspect of myosmine uptake by humans from other biological sources and subsequent biological activation. The new reaction pathway indicates that human exposure to nicotinoid nitrosation products seems to be not restricted exclusively to tobacco. PMID- 10691646 TI - Phase behavior and component partitioning in low water content amorphous carbohydrates and their potential impact on encapsulation of flavors. AB - The compositions at which amorphous ethanol-maltose-water mixtures exhibit liquid liquid separation have been determined in the temperature range from 20 to 80 degrees C. At water contents below approximately 20% w/w two phases were observed, with the maltose-rich phase slightly richer in water. Partition coefficients of organic nonelectrolytes ranging in hydrophobicity from 1, 2 ethanediol and 1,2-propanediol to benzyl alcohol and propyl acetate have been measured for octanol/sorbitol, benzyl alcohol/sorbitol, and 1-butanol/sorbitol mixtures. Linear correlations were found between the log partition coefficients in the various solvent systems. Replacing water with sorbitol results in more organic partitioning into the octanol. Replacing octanol with benzyl alcohol or 1 butanol also results in more organic partitioning into the hydrophobic phase. The results establish a relationship with partition coefficients for octanol/water mixtures, which are well studied experimentally and for which predictive approaches exist. The implications of these results for flavor retention and encapsulation are discussed. PMID- 10691647 TI - Identification of impact odorants in Bordeaux red grape juice, in the commercial yeast used for its fermentation, and in the produced wine. AB - The aroma extract dilution analysis method was used to detect the impact odorants of Bordeaux Cabernet Sauvignon and Merlot wines extracts, as well as those of the extracts of the corresponding Cabernet Sauvignon juice and dry yeasts used for its fermentation. The wines and the yeasts were extracted using dichloromethane, and the juice was extracted using Amberlite XAD-2. Structural identification of the impact odorants using gas chromatography-mass spectrometry and atomic emission detection (sulfur acquisition) was achieved after enrichment of these extracts by silica gel and Affi-Gel 501 chromatography. The same odorants (with the exception of dimethyl sulfide among 48) were detected in both wine extracts, with about the same flavor dilution (FD) factors. The 18 impact odorants detected in the Cabernet Sauvignon juice and dry yeast extracts were also found in the wine extracts. The odorants with the highest FD factors were 3 (methylsulfanyl)propanal, (E,Z)-nona-2, 6-dienal, and decanal in the juice extract, 2-methyl-3-sulfanylfuran, 3-(methylsulfanyl)propanal, 2-/3 methylbutanoic acids, and phenylethanal in the dry yeast extract, and 2-/3 methylbutanols, 2-phenylethanol, 2-methyl-3-sulfanylfuran, acetic acid, 3 (methylsulfanyl)propanal, 2-/3-methylbutanoic acids, beta-damascenone, 3 sulfanylhexan-1-ol, Furaneol, and homofuraneol in the wine extracts. Determination of the odor thresholds of some of these impact odorants was carried out. PMID- 10691649 TI - Volatile compounds from Escherichia coli O157:H7 and their absorption by strawberry fruit. AB - Volatile compounds emitted by cultures of two strains of the pathogenic bacterium Escherichia coliO157:H7 and a nonpathogenic strain of E. coli were trapped on Super-Q porous polymer and identified by GC-MS. The predominant compound produced by all three strains was indole with lesser amounts of other components including methyl ketones, 2-heptanone, 2-nonanone, 2-undecanone, and 2-tridecanone. The vapor-phase profiles of these strains were similar for most chemicals identified but differed with regard to ketones. Strawberry fruit was shown to be a suitable host for E. coli O157:H7 with the population of the bacterium either increasing or remaining stable after 3 days depending on inoculation level. Headspace analysis of the volatile compounds from inoculated fruit yielded no detectable quantity of indole. Strawberry fruit readily absorbed indole and other volatile compounds produced by the bacteria and in some cases metabolized the compounds to new volatile products. Thus, headspace "marker" compounds indicating possible bacterial contamination of fruit were largely removed from the vapor phase by the strawberries. PMID- 10691648 TI - Study of interactions between food phenolics and aromatic flavors using one- and two-dimensional (1)H NMR spectroscopy. AB - Changes in flavor release and aroma characteristics in a medium including food phenolics may be attributed to an intermolecular interaction between flavor compounds and phenolics. To investigate the interaction, one- and two-dimensional NMR studies have been carried out on the binding of two phenolics, gallic acid and naringin, with three aroma compounds, 2-methylpyrazine, vanillin, and ethyl benzoate. Evaluation of thermodynamic parameters and intermolecular nuclear Overhauser effects reveals that gallic acid can interact more strongly with aromatic flavors than naringin. The supramolecular complexation is also dependent on the structural nature of the flavors, with 2-methylpyrazine and vanillin interacting more strongly than ethyl benzoate. The interaction is principally pi pi stacking between the galloyl ring and the aromatic ring of the aroma compounds, but secondary hydrogen-bonding effects help to stabilize the complex and enhance the specificity. PMID- 10691650 TI - 3-Mercapto-2-methylpentan-1-ol, a new powerful aroma compound. AB - 3-Mercapto-2-methylpentan-1-ol was first detected in a complex thermally processed flavor and finally isolated from raw onions. The chemical structure of this new compound was identified by MS and (1)H NMR measurement and synthesis of the proposed structure. Sensory evaluation at different concentrations indicated that the flavor quality is strongly dependent on concentration. At low concentration (0.5 ppb) a pleasant meat broth, sweaty, onion, and leek-like odor can be perceived. On the basis of some isolation experiments and volatiles occurring in raw onions, a formation pathway is proposed. As one intermediate 3 mercapto-2-methylpentanal, another new strong flavor compound, was suggested. The presence of this compound in raw onions was confirmed by synthesis and comparison of MS and chromatographic data. PMID- 10691651 TI - Enantioselective syntheses and sensory properties of the 3-mercapto-2 methylpentanols. AB - 3-Mercapto-2-methylpentanol, a new powerful flavor compound, exhibits two stereocenters giving rise to two pairs of diastereomers. To determine differences in the sensory properties, all four diastereomers and enantiomers were stereo- and enantioselectively synthesized. A highly diastereoselective aldol reaction using a chiral auxiliary was one of the key steps in the synthesis. Further derivatization yielded the enantiopure compounds. Odor thresholds in air and water were determined. PMID- 10691652 TI - Distribution of S-Alk(en)ylcysteine sulfoxides in some Allium species. Identification Of a new flavor precursor: S-ethylcysteine sulfoxide (Ethiin). AB - The content of S-alk(en)ylcysteine sulfoxides, important nonvolatile flavor precursors, was determined in 15 different Allium species by means of gas chromatography. The method employed is based on derivatization of S alk(en)ylcysteine sulfoxides with ethyl chloroformate followed by their reduction with sodium iodide. The total content of S-alk(en)ylcysteine sulfoxides varied considerably in the wide range between 0.02 and 1.3% fresh weight. Not only the total content but also relative proportions of individual derivatives varied to a great extent. A novel S-alkylcysteine derivative, S-ethylcysteine sulfoxide (ethiin), not previously reported to occur in Allium species, was found in most of the samples examined in trace amounts. None of the other S-alk(en)ylcysteine sulfoxides, for example, isopropyl, (Z)-1-propenyl, butyl, or pentyl, were detected in any of the samples analyzed, limiting possible levels of each of these components to 0. 0006). This was attributed to the coextraction of cross-reactive SDM related residues that were not quantified by the HPLC method. The presence of these residues should be considered during data interpretation when ELISA methods coupled with rapid aqueous extraction of samples are used in SDM residue monitoring programs. PMID- 10691674 TI - Effect of temperature and/or pressure on tomato pectinesterase activity. AB - The activity of tomato pectinesterase (PE) was studied as a function of pressure (0.1-900 MPa) and temperature (20-75 degrees C). Tomato PE was rather heat labile at atmospheric pressure (inactivation in the temperature domain 57-65 degrees C), but it was very pressure resistant. Even at 900 MPa and 60 degrees C the inactivation was slower as compared to the same treatment at atmospheric pressure. At atmospheric pressure, optimal catalytic activity of PE was found at neutral pH and a temperature of 55 degrees C. Increasing pressure up to 300 MPa increased the enzyme activity as compared to atmospheric pressure. A maximal enzyme activity was found at 100-200 MPa combined with a temperature of 60-65 degrees C. The presence of Ca(2+) ions (60 mM) decreased the enzyme activity at atmospheric pressure in the temperature range 45-60 degrees C but increased enzyme activity at elevated pressure (up to 300 MPa). Maximal enzyme activity in the presence of Ca(2+) ions was noted at 200-300 MPa in combination with a temperature of 65-70 degrees C. PMID- 10691675 TI - Effect of gamma-irradiation on phenolic compounds and phenylalanine ammonia-lyase activity during storage in relation to peel injury from peel of Citrus clementina hort. Ex. tanaka. AB - The influence of gamma-irradiation on the content of phenolic compounds was evaluated on Moroccan Citrus fruits (Citrus clementina Hort. ex. Tanaka) treated at a mean dose of 0.3 kGy and stored for 49 days at 3 degrees C. The results show that irradiation has enhanced the synthesis of total phenolic compounds and is correlated with phenylalanine ammonia-lyase activity (PAL) during storage. Accumulation of phenolic compounds in cells is demonstrated and may be explained by the enhancement of PAL activity. HPLC/UV (diode array detector) analysis demonstrated that hesperidin was the major flavanone and nobiletin and heptamethoxyflavone were the major polymethoxylated flavones. Hesperidin is also the major phenolic compound in clementines. Irradiation stimulates the biosynthesis of hesperidin after 14 days of storage, corresponding to the maximum of PAL activity. p-Coumaric acid was also identified, and its content was particularly high in irradiated fruits after 49 days of storage. Accumulation of flavonoids and p-coumaric acid could be related to a better resistance. The percentage of losses due to peel injury "pitting" during storage was between 1 and 5% after 49 days of storage. The connections between irradiation, enzyme activity, phenolic content, and peel injury are briefly discussed. PMID- 10691676 TI - Specificity of papaya lipase in esterification with respect to the chemical structure of substrates. AB - Esterification, catalyzed by papaya (Carica papaya) lipase (CPL), was studied with various alcohols and carboxylic acids under competitive conditions. Acids studied were straight-chain saturates of different chain lengths, with octanoic acid as the reference. Alcohols chosen were aliphatic straight-chain, branched, secondary, tertiary, terpene, and aromatic alcohols of different chain lengths, using 1-hexanol as the reference. The initial reaction rate increased with increasing chain length of the acid from C4:0 to C18:0, followed by a slight decrease with C20:0. In the case of alcohols, an optimum chain length of 8 carbon atoms was obtained for the straight-chain aliphatic group (C2 to C16). Ethanol, 1 propanol, and secondary and tertiary alcohols showed rather low reactivity. Branching of the alcohols was found not to affect the reactivity in esterification; among the terpenes, beta-citronellol [(2E)-3, 7-dimethyl-6 octenol] and geraniol [(2E)-3,7-dimethylocta-2, 6-dien-1-ol] were found to be more reactive than nerol [(2Z)-3, 7-dimethylocta-2,6-dien-1-ol]. The highest reaction rate was found for the aromatic benzyl alcohol (phenylmethanol). PMID- 10691677 TI - Immunological characterization of recombinant soy protein allergen produced by Escherichia coli expression system. AB - To elucidate the molecular mechanism of the allergenicity of soybean P34 protein recognized as the most allergenic protein in soybean, the protein was expressed in Escherichia coli transformed with a plasmid carrying P34 cDNA. SDS-PAGE pattern showed that the molecular weight of the recombinant P34 was approximately 2 kDa less than that of the native soybean P34. The difference in the molecular mass between these two proteins could be due to the native P34 in soybean being glycosylated at position Asn(170), whereas the recombinant protein generated in E. coli lacks this post-translational modification. Immunoblot analysis showed that both soybean and recombinant P34 proteins cross-reacted not only with polyclonal and monoclonal antibodies produced against P34 and crude soybean protein but also with patients' sera. The results suggest that the recombinant P34 is immunologically reactive, indicating that both proteins have similar epitope structures. Thus, the recombinant P34 produced by the E. coli expression system can be used as a standard allergen for molecular design to reduce the allergenic structure. PMID- 10691678 TI - Interaction of curcumin with phosphatidylcholine: A spectrofluorometric study PMID- 10691679 TI - Structure-physicochemical function relationships of soybean beta-conglycinin constituent subunits PMID- 10691680 TI - The PPARs: from orphan receptors to drug discovery. PMID- 10691681 TI - Novel ligands lacking a positive charge for the delta- and mu-opioid receptors. AB - Recently we reported using minilibraries to replace Lys(9) [somatostatin (SRIF) numbering] of the potent somatostatin agonist L-363,301 (c[-Pro-Phe-D-Trp-Lys-Thr Phe-]) to generate the potent neurokinin receptor (NK-1) antagonist c[-Pro-Phe-D Trp-p-F-Phe-Thr-Phe-]. This novel cyclic hexapeptide did not bind the SRIF receptor. Thus, a single mutation converted L-363,301, a SRIF agonist with potency ca. 2-8 times the potency of SRIF in laboratory animals,(24) into a selective NK-1 receptor antagonist with an IC(50) of 2 nM in vitro. During the screening of the same libraries for ligands of the delta-opioid receptor, we identified four compounds (1-4) which represent a new class of delta-opioid antagonists, some of which were also NK-1 receptor antagonists. The most potent delta-opioid antagonist, c[-Pro-1-Nal-D-Trp-Tyr-Thr-Phe-] (2), showed a K(e) value of 128 nM in the mouse vas deferens assay and a delta-receptor binding affinity constant of 152 nM in the rat brain membrane binding assay. These results are of interest because they represent a novel class of delta-opioid antagonists and, like two previously reported delta-opioid antagonists, they lack a positive charge. To examine further the requirement for a positive charge in the delta-opioid ligands, we prepared two analogues of the beta-casomorphin derived mixed mu-agonist/delta-antagonist, H-Dmt-c[-D-Orn-2-Nal-D-Pro-Gly-] (7), in which we eliminated the positive charge either through formylation of the primary amino group (5) or by the deletion of this N-terminal amino group (6). These latter compounds proved to be delta-opioid antagonists with K(e) values in the 16-120 nM range, as well as fairly potent mu-opioid antagonists (K(e) approximately 200 nM). These six compounds provide the most convincing evidence to date that there is no requirement for a positive charge in mu- and delta opioid receptor antagonists. In addition, cyclic hexapeptide 4 lacks a phenolic hydroxyl group. Taken together, these data suggest that the prevailing assumptions about delta- and mu-opioid receptor binding need revision and that the receptors for these opioid ligands have much in common with the NK-1 and somatostatin receptors. PMID- 10691682 TI - Phenyl beta-methoxyacrylates: a new antimalarial pharmacophore. AB - Phenyl beta-methoxyacrylates, linked to an aromatic ring via an olefinic bridge, have been identified as novel, potentially inexpensive, antimalarial agents. The compounds are believed to exert their activity by inhibition of mitochondrial electron transport at the cytochrome bc(1) complex. A series of compounds have been synthesized to define structure-activity relationships affecting antimalarial activity. It was found that the beta-methoxyacrylate was required ortho to the linker and the optimal bridge was (E,E)-butadiene. Compounds in which the second aromatic ring was ortho-substituted or ortho,para-disubstituted gave optimal potency. Several compounds were identified with potency that is superior to that of chloroquine both in culture and in a murine malaria model. PMID- 10691683 TI - Opiate aromatic pharmacophore structure-activity relationships in CTAP analogues determined by topographical bias, two-dimensional NMR, and biological activity assays. AB - Topographically constrained analogues of the highly mu-opioid-receptor-selective antagonist CTAP (H-D-Phe-c[Cys-Tyr-D-Trp-Arg-Thr-Pen]-Thr-NH(2), 1) were prepared by solid-phase peptide synthesis. Replacement of the D-Phe residue with conformationally biased beta-methyl derivatives of phenylalanine or tryptophan (2R,3R; 2R,3S; 2S,3R; 2S,3S) yielded peptides that displayed widely varying types of biological activities. In an effort to correlate the observed biological activities of these analogues with their structures, two-dimensional (1)H NMR and molecular modeling was performed. Unlike the parent (1), which is essentially a pure mu antagonist with weak delta agonist activities in the MVD bioassay, the diastereomeric beta-MePhe(1)-containing peptides exhibited simultaneous delta agonism and mu antagonism by the (2R,3R)-containing isomer 2; mu antagonism by the (2R,3S)-containing isomer 3; weak mu agonism by the (2S,3R)-containing isomer 4; and delta agonism by the (2S,3S)-containing isomer 5. Incorporation of beta MeTrp isomers into position 1 led to peptides that were mu antagonists (2R,3R), 8; (2R,3S), 9, or essentially inactive (<10%) in the MVD and GPI assays (2S,3R), 10; (2S,3S), 11. Interestingly, in vivo antinociceptive activity was predicted by neither MVD nor GPI bioactivity. When D-Trp was incorporated in position 1, the result (7) is a partial, yet relatively potent mu agonist which also displayed weak delta agonist activity. Molecular modeling based on 2D NMR revealed that low energy conformers of peptides with similar biological activities had similar aromatic pharmacophore orientations and interaromatic distances. Peptides that exhibit mu antagonism have interaromatic distances of 7.0-7.9 A and have their amino terminal aromatic moiety pointing in a direction opposite to the direction that the amino terminus points. Peptides with delta opioid activity displayed an interaromatic distance of <7 A and had their amino terminal aromatic moiety pointing in the same direction as the amino terminus. PMID- 10691684 TI - Chiral nonsteroidal affinity ligands for the androgen receptor. 1. Bicalutamide analogues bearing electrophilic groups in the B aromatic ring. AB - A series of chiral analogues of bicalutamide bearing electrophilic groups (isothiocyanate, N-chloroacetyl, and N-bromoacetyl) on aromatic ring B of the parent molecule were synthesized. These compounds were designed as affinity ligands for the androgen receptor (AR). We prepared the (R)- and (S)-optical isomers of these compounds as pure enantiomers. The AR binding affinities of these compounds were measured in a competitive binding assay with the radiolabeled high-affinity AR ligand, [(3)H]mibolerone. In accordance with our previous results for the enantiomers of bicalutamide, we found that all (R) isomers demonstrated much higher binding affinity to the AR as compared to their corresponding (S)-isomers. The para-substituted affinity ligands in ring B bound the AR with higher affinities than the corresponding meta-substituted analogues. Oxidation of thioester affinity ligands to their sulfonyl analogues for the para substituted compounds decreased AR binding affinities and similar modification increased binding affinities for corresponding meta-analogues. The least potent para-substituted sulfonyl compounds had higher AR binding affinities than the most potent meta-substituted sulfonyl compounds. Overall, the para-substituted unoxidized molecules demonstrated the highest AR binding affinity. Subsequent research using AR exchange assays and Scatchard analyses showed that the isothiocyanate affinity ligands (R)-7, (R)-9, and (R)-10 reported herein are the first specific chemoaffinity ligands for the AR. PMID- 10691685 TI - Synthesis and human beta-adrenoceptor activity of 1-(3,5-diiodo-4- methoxybenzyl) 1,2,3,4-tetrahydroisoquinolin-6-ol derivatives in vitro. AB - Trimetoquinol (1, TMQ) is a potent nonselective beta-adrenergic receptor (AR) agonist and a thromboxane A(2)/prostaglandin endoperoxide (TP) receptor antagonist, while 3',5'-diiodo-TMQ (2) exhibits beta(3)-AR selectivity. In search of selective beta(3)-AR agonists as potential drugs for the treatment of human obesity and type II diabetes mellitus, a series of 1-(3, 5-diiodo-4 methoxybenzyl)-1,2,3,4-tetrahydroisoquinolin-6-ols has been prepared and evaluated for their biological activities at human beta(1)-, beta(2)-, and beta(3)-ARs expressed in Chinese hamster ovary (CHO) cells. The compounds have been synthesized by the Bischler-Napieralski cyclization of corresponding amides followed by NaBH(4) reduction, and the halogens in the aromatic ring A were introduced by direct halogenation of protected compound 11. Whereas halogen substitution in ring A reduced either potency or intrinsic activity on beta(3) AR, the non-halogen-substituted compounds 8 and 10 were potent, selective, nearly full agonists for beta(3)-AR. PMID- 10691686 TI - Binding and preliminary evaluation of 5-hydroxy- and 10-hydroxy-2,3, 12,12a tetrahydro-1H-[1]benzoxepino[2,3,4-ij]isoquinolines as dopamine receptor ligands. AB - The N-methyl, N-ethyl, and N-n-propyl derivatives of 5-hydoxy- and 10-hydroxy 2,3,12,12a-tetrahydro-1H-[1]benzoxepino[2,3, 4-ij]isoquinolines were prepared as monophenolic ligands for the dopamine receptor and evaluated for their affinity at D(1)-like and D(2)-like subtypes. All compounds showed very low D(1) affinities. This could be ascribed to the absence of a catechol nucleus or of the beta-phenyldopamine pharmacophore. Only the N-methyl-5-hydroxy- (5a), N-methyl-10 hydroxy- (6a), and N-methyl-4-bromo-10-methoxy-2,3, 12,12a-tetrahydro-1H [1]benzoxepino[2,3,4-ij]isoquinolines (26a) bound the D(2) receptors with low affinity, in the same range as dopamine. In compounds 5a and 6a, the 2-(3 hydroxyphenyl)ethylamine moiety does not meet the requirements of the D(2) agonist pharmacophore: namely, the 2-(3-hydroxyphenyl)ethylamine does not reach the trans, fully extended conformation. The three compounds did not interact with recombinant human D(4) receptors, and only 5a showed low affinity for rat recombinant D(3) receptors. Analysis of the influence of Na(+) on [(3)H]spiperone binding showed that 5a displays a potential dopamine D(2) agonist profile, whereas 6a probably has a dopamine D(2) antagonist activity. The D(2) agonist activity of 5a was proved by the effects on prolactin release from primary cultures of rat anterior pituitary cells. PMID- 10691687 TI - Hypocholesterolemic and hypolipidemic activity of some novel morpholine derivatives with antioxidant activity. AB - In this investigation, we study the synthesis and the evaluation of antioxidant and hypocholesterolemic activity of a number of 2-biphenylyl morpholine derivatives, which are structurally similar to some substituted morpholines possessing antioxidant activity, as well as to hypocholesterolemic 3-biaryl quinuclidines. The novel derivatives are found to inhibit the ferrous/ascorbate induced lipid peroxidation of microsomal membrane lipids, the most potent derivative, 2-(4-biphenyl)-4-methyl-octahydro-1,4-benzoxazin-2-ol (compound 7), having an IC(50) value of 250 microM. In addition, these compounds demonstrate hypocholesterolemic and hypolipidemic action. The most active compound (7) decreases total cholesterol, low density lipoprotein, and triglycerides in plasma of Triton WR-1339 induced hyperlipidemic rats by 54%, 51%, and 49%, respectively, at 28 micromol/kg (ip). The above results indicate that the new molecules may be proven useful as leads for the design of novel compounds as potentially antiatherogenic factors. PMID- 10691688 TI - Steroidal affinity labels of the estrogen receptor alpha. 4. Electrophilic 11beta aryl derivatives of estradiol. AB - Ten electrophilic estradiol 11beta-aryl derivatives were synthesized, with three different types of 11beta-substituent: (i) pOO(CH(2))(2)X (compounds: 6, X = OSO(2)CH(3); 7, X = I; 13, X = NHCOCH(2)Cl; 15, X = N(CH(3))COCH(2)Br; and 16, X = N(CH(3))COCH(2)Cl); (ii) pOO(CH(2))(5)X (compounds: 17, X = I; 20, X = NHCOCH(2)Br; and 22, X = N(CH(3))COCH(2)Br); and (iii) pOC(triple bond)CCH(2)X (compounds: 27, X = NHCOCH(2)Cl; and 29, X = N(CH(3))COCH(2)Cl). The range of their apparent affinity constants for binding the lamb uterine estrogen receptor alpha (ERalpha) was 3-40% that of estradiol. Six electrophiles, chloroacetamides 13, 16, 27, and 29, iodide 17, and bromoacetamide 20 (whose arm linking the electrophilic carbon to the 11beta-phenyl group includes at least six bonds), were able to irreversibly inhibit the binding of [(3)H]estradiol to ER (25-60% decrease in binding sites), with the following compound effectiveness order: 17 < 13 < 16 approximately 20 approximately 27 approximately 29. Mesylate 6, iodide 7 (whose linking arm includes only three bonds), and bromoacetamides 15 and 22 (which differ from 16 by the Cl to Br change and from 20 by the NH to NCH(3) change, respectively) were much less effective (<10% decrease in binding sites, if any). The fact that the inactivation of estradiol-binding sites by the six electrophiles was totally prevented by estradiol indicated that they were ER affinity labeling agents. When ER was modified by methyl methanethiosulfonate, an SH-specific reagent, the different compounds led to very contrasting results in ER affinity labeling. With modified ER, iodide 17 and chloroacetamides 27 and 29 were practically inactive, chloroacetamides 13 and 16 and bromoacetamide 20 were still active but less effective than on the native ER, whereas tertiary bromoacetamides 15 and 22, found to be practically inactive on native ER, became the most effective electrophiles ( approximately 45% and approximately 65% binding sites inactivated, respectively). The results indicate that in the steroid-filled hormone-binding pocket: (i) nucleophilic residues are localized on the beta-side but relatively remote from the steroid nucleus (distance from C-11 > "seven bonds"); (ii) relatively discrete changes in the electrophilic functionality, such as Cl to Br or NH to NCH(3) of haloacetamido compounds, can markedly modify the positioning of the electrophilic center which could no longer react with the nucleophilic residues; and (iii) cysteine residues (probably homologues of human ERalpha cysteine 381 and/or cysteine 530) are, at least partly, the covalent attachment sites of the electrophiles. Moreover, modification of cysteine residues by methyl methanethiosulfonate changes the structure of the hormone-binding pocket, whose labeling by the various electrophiles is profoundly altered. PMID- 10691689 TI - Inhibitors of topoisomerase II based on the benzodiimidazole and dipyrroloimidazobenzimidazole ring systems: controlling DT-diaphorase reductive inactivation with steric bulk. AB - Described herein are the synthesis, cytotoxic properties, and topoisomerase II inhibition assays of benzodiimidazole and dipyrroloimidazobenzimidazole structural variants of the pyrrolo[1, 2-a]benzimidazole or APBI ring system. These ring variants were designed to inhibit topoisomerase II, much as the APBIs are able to do. Since only the quinone form of the APBIs can intercalate DNA, two electron reduction to the hydroquinone by DT-diaphorase is known to deactivate these compounds. Indeed, the APBIs possess a high inverse correlation with the cellular concentration of DT-diaphorase. Therefore one feature of the ABPI structural variants is the excessive bulk about the quinone ring, which was predicted to diminish DT-diaphorase substrate activity. Another feature is the presence of one or two alkylating centers, which would permit alkylation of DNA and/or topoisomerase II. Inhibition assays for topoisomerase II-mediated relaxation of supercoiled DNA indicate that the benzodiimidazole and dipyrroloimidazobenzimidazole quinone ring systems are catalytic inhibitors of topoisomerase II. Both quinone systems exhibit cytotoxicity perhaps due to the lack of inactivation by DT-diaphorase as well as topoisomerase II inhibition. One quinone displayed the novel feature of cytotoxicity selectively against melanoma cell lines. In conclusion, the benzodiimidazole and dipyrroloimidazobenzimidazole quinone ring systems will be subjected to future analogue development and structure-activity studies. PMID- 10691690 TI - Synthesis, biodistribution, and primate imaging of fluorine-18 labeled 2beta carbo-1'-fluoro-2-propoxy-3beta-(4-chlorophenyl)tr opanes. Ligands for the imaging of dopamine transporters by positron emission tomography. AB - 2beta-(R)-Carbo-1-fluoro-2-propoxy-3beta-(4-chlorophenyl) tro pane ((R)-FIPCT, R 6) and 2beta-(S)-carbo-1-fluoro-2-propoxy-3beta-(4-chlorophenyl) tro pane ((S) FIPCT, S-6) were prepared and evaluated in vitro and in vivo for dopamine transporter (DAT) selectivity and specificity. High specific activity [(18)F](R) FIPCT and [(18)F](S)-FIPCT were synthesized in 5% radiochemical yield (decay corrected to end of bombardment (EOB)) by preparation of the precursors 2beta carbo-R-1-mesyloxy-2-propoxy-3beta-(4-chlorop hen yl)tropane (R-12) and 2beta carbo-S-1-mesyloxy-2-propoxy-3beta-(4-chlorop hen yl)tropane (S-12) followed by treatment with no carrier-added potassium[(18)F]fluoride and kyrptofix K222 in acetonitrile. Competition binding in cells stably expressing the transfected human DAT and serotonin transporter (SERT) labeled by [(3)H]WIN 35428 and [(3)H]citalopram, respectively, demonstrated the following order of DAT affinity (K(i) in nM): GBR 12909 (0.36) > CIT (0.48) > (S)-FIPCT (0.67) >> (R)-FIPCT (3.2). The affinity of (S)-FIPCT and (R)-FIPCT for SERT was 127- and 20-fold lower, respectively, than for DAT. In vivo biodistribution studies were performed in male rats and demonstrated that the brain uptake of [(18)F](R)-FIPCT and [(18)F](S)-FIPCT were selective and specific for DAT rich regions (caudate and putamen). PET brain imaging studies in monkeys demonstrated high [(18)F](R)-FIPCT and [(18)F](S)-FIPCT uptake in the caudate and putamen which resulted in caudate to-cerebellum and putamen-to-cerebellum ratios of 2.5-3.5 at 115 min. [(18)F](R) FIPCT uptake in the caudate/putamen achieved transient equilibrium at 75 min. In an imaging experiment with [(18)F](S)-FIPCT in a rhesus monkey with its left hemisphere lesioned with MPTP, radioactivity was reduced to background in the caudate and putamen of the lesioned hemisphere. The high specific activity one step radiolabeling preparation and high specificity and selectivity of [(18)F](R) FIPCT and [(18)F](S)-FIPCT for DAT indicate [(18)F](R)-FIPCT and [(18)F](S)-FIPCT are potential radioligands for mapping brain DAT in humans using PET. PMID- 10691691 TI - Diamino benzo[b]thiophene derivatives as a novel class of active site directed thrombin inhibitors. 5. Potency, efficacy, and pharmacokinetic properties of modified C-3 side chain derivatives. AB - A systematic investigation of the structure-activity relationships of the C-3 side chain of the screening hit 1a led to the identification of the potent thrombin inhibitors 23c, 28c, and 31c. Their activities (1240, 903, and 1271 x 10(6) L/mol, respectively) represent 2200- and 2900-fold increases in potency over the starting lead 1a. This activity enhancement was accomplished with an increase of thrombin selectivity. The in vitro anticoagulant profiles of derivatives 28c and 31c were determined, and they compare favorably with the clinical agent H-R-1-[4aS, 8aS]perhydroisoquinolyl-prolyl-arginyl aldehyde (D-Piq Pro-Arg-H; 32). The more potent members of this series have been studied in an arterial/venous shunt (AV shunt) model of thrombosis and were found to be efficacious in reducing clot formation. However, their efficacy is currently limited by their rapid and extensive distribution following administration. PMID- 10691692 TI - Inhibitors of tripeptidyl peptidase II. 2. Generation of the first novel lead inhibitor of cholecystokinin-8-inactivating peptidase: a strategy for the design of peptidase inhibitors. AB - The cholecystokinin-8 (CCK-8)-inactivating peptidase is a serine peptidase which has been shown to be a membrane-bound isoform of tripeptidyl peptidase II (EC 3.4.14.10). It cleaves the neurotransmitter CCK-8 sulfate at the Met-Gly bond to give Asp-Tyr(SO(3)H)-Met-OH + Gly-Trp-Met-Asp-Phe-NH(2). In seeking a reversible inhibitor of this peptidase, the enzymatic binding subsites were characterized using a fluorimetric assay based on the hydrolysis of the artificial substrate Ala-Ala-Phe-amidomethylcoumarin. A series of di- and tripeptides having various alkyl or aryl side chains was studied to determine the accessible volume for binding and to probe the potential for hydrophobic interactions. From this initial study the tripeptides Ile-Pro-Ile-OH (K(i) = 1 microM) and Ala-Pro-Ala-OH (K(i) = 3 microM) and dipeptide amide Val-Nvl-NHBu (K(i) = 3 microM) emerged as leads. Comparison of these structures led to the synthesis of Val-Pro-NHBu (K(i) = 0.57 microM) which served for later optimization in the design of butabindide, a potent reversible competitive and selective inhibitor of the CCK-8-inactivating peptidase. The strategy for this work is explicitly described since it illustrates a possible general approach for peptidase inhibitor design. PMID- 10691693 TI - Palladium-catalyzed cross-coupling reactions for the synthesis of 6, 8 disubstituted 1,7-naphthyridines: a novel class of potent and selective phosphodiesterase type 4D inhibitors. AB - Recently, four subtypes of the human phosphodiesterase type 4 (PDE4A-D) enzyme have been described. So far, only very few PDE4 subtype-selective inhibitors are known. Herein, we describe the synthesis of 6,8-disubstituted 1,7-naphthyridines and their characterization as potent and selective inhibitors of PDE4D which suppress the oxidative burst in human eosinophils with IC(50) values as low as 0.7 nM. SAR development and the extended use of palladium-catalyzed cross coupling reactions led to compound 11 which inhibited human PDE4D with an IC(50) value of 1 nM. Thus, compound 11 was 55, 175, and 1000 times more potent in inhibiting PDE4D over PDE4B, PDE4A, and PDE4C. In a Brown Norway rat model of allergic asthma, compound 11 when given by the oral route (1 mg/kg) reduced by more than 50% the influx of eosinophils, T-cells, and neutrophils into bronchoalveolar lavage fluid (BALF) samples obtained from antigen-challenged animals. Thus, PDE4D-selective inhibitors of the 1,7-naphthyridine class have the potential as an oral therapy for treating asthma. PMID- 10691694 TI - Benzyl derivatives of 2,1,3-benzo- and benzothieno[3,2-a]thiadiazine 2,2 dioxides: first phosphodiesterase 7 inhibitors. AB - The synthesis of a new family of benzyl derivatives of 2,1,3-benzo- and benzothieno[3,2-a]thiadiazine 2,2-dioxides was achieved. The biological data revealed the first heterocyclic family of compounds with PDE 7 inhibitory properties appearing to be a new objective for the treatment of T-cell-dependent disorders. The IC(50) values or percent inhibition values of the compounds against PDE 7 were calculated by testing them against human recombinant PDE 7 expressed in S. cerevisiae. In this expression system the only cyclic nucleotide hydrolyzing activity present in cell extracts corresponded to human PDE 7. Isoenzyme selectivity PDE 7 versus PDE 4 and PDE 3 was also measured. Considering simultaneously inhibition of the three different isoenzymes, monobenzyl derivatives 15 and 23 showed interesting PDE 7 potency (around 10 microM); although not statistically significant, a trend toward selectivity with respect to PDE 3 and PDE 4 was obtained. Benzothiadiazine 16, although less potent at PDE 7 (IC(50) = 25 microM), also showed a trend of selectivity toward PDE 3 and PDE 4. These compounds are considered the best leads for further optimization. PMID- 10691695 TI - Heteroarylmethoxyphenylalkoxyiminoalkylcarboxylic acids as leukotriene biosynthesis inhibitors. AB - A novel series of heteroarylmethoxyphenylalkoxyiminoalkylcarboxylic acids was studied as leukotriene biosynthesis inhibitors. A hypothesis of structure activity optimization by insertion of an oxime moiety was investigated using REV 5901 as a starting point. A systematic structure-activity optimization showed that the spatial arrangement and stereochemistry of the oxime insertion unit proved to be important for inhibitory activity. The promising lead, S-(E)-11, inhibited LTB(4) biosynthesis in the intact human neutrophil with IC(50) of 8 nM and had superior oral activity in vivo, in a rat pleurisy model (ED(50) = 0.14 mg/kg) and rat anaphylaxis model (ED(50) = 0.13 mg/kg). In a model of lung inflammation, S-(E)-11 blocked LTE(4) biosynthesis (ED(50) of 0.1 mg/kg) and eosinophil influx (ED(50) of 0.2 mg/kg). S-(E)-11 (A-93178) was selected for further preclinical evaluation. PMID- 10691696 TI - A new class of conformationally rigid analogues of 4-amino-5-halopentanoic acids, potent inactivators of gamma-aminobutyric acid aminotransferase. AB - Recently, we found (Qiu, J.; Pingsterhaus, J. M.; Silverman, R. B. J. Med. Chem. 1999, 42, 4725-4728) that conformationally rigid analogues of the GABA aminotransferase (GABA-AT) inactivator vigabatrin were not inactivators of GABA AT. To determine if this is a general phenomenon of GABA-AT inactivators, several mono- and di-halogen-substituted conformationally rigid analogues (7-15) of other GABA-AT inactivators, 4-amino-5-halopentanoic acids, were synthesized as potential inactivators of GABA-AT. Four of them, (+)-7, (-)-9, (+)-10, and (+) 15, were inactivators, although not as potent as the corresponding open-chain analogues. The maximal inactivation rate constants, k(inact), for the fluoro- and bromo-substituted analogues were comparable, indicating that cleavage of the C-X bond is not rate determining. Consistent with that observation is the finding that [3-(2)H]-10 exhibits a deuterium isotope effect on inactivation of 3.3, suggesting that C-H bond cleavage is the rate-determining step. The rate of inactivation of GABA-AT by the fluorinated analogue 7 is 1/15 that of inactivation by the corresponding open-chain analogue, 4-amino-5-fluoropentanoic acid (3a). Whereas inactivation by 3a releases only one fluoride ion, inactivation by 7 releases 148 fluoride ions, accounting for the less efficient inactivation rate. Inactivation leads to covalent attachment of 2 equiv of inactivator after gel filtration; upon urea denaturation, 1 equiv of radioactivity remains bound to the enzyme. This suggests that, unlike the open chain anlogue, the conformationally rigid analogue becomes, at least partially, attached to an active-site residue. It appears that the conformational constraint has a larger effect on inactivators that inactivate by a Michael addition mechanism than by an enamine mechanism. PMID- 10691697 TI - Structure-activity relationship studies on 1-[2-(4 Phenylphenoxy)ethyl]pyrrolidine (SC-22716), a potent inhibitor of leukotriene A(4) (LTA(4)) hydrolase. AB - Leukotriene B(4) (LTB(4)) is a pro-inflammatory mediator that has been implicated in the pathogenesis of a number of diseases including inflammatory bowel disease (IBD) and psoriasis. Since the action of LTA(4) hydrolase is the rate-limiting step for LTB(4) production, this enzyme represents an attractive pharmacological target for the suppression of LTB(4) production. From an in-house screening program, SC-22716 (1, 1-[2-(4-phenylphenoxy)ethyl]pyrrolidine) was identified as a potent inhibitor of LTA(4) hydrolase. Structure-activity relationship (SAR) studies around this structural class resulted in the identification of a number of novel, potent inhibitors of LTA(4) hydrolase, several of which demonstrated good oral activity in a mouse ex vivo whole blood assay. PMID- 10691698 TI - The cyclohexene ring system as a furanose mimic: synthesis and antiviral activity of both enantiomers of cyclohexenylguanine. AB - Both enantiomers of cyclohexenylguanine were synthesized in a stereospecific way starting from the same starting material: R-(-)-carvone. Both compounds showed potent and selective anti-herpesvirus activity (HSV-1, HSV-2, VZV, CMV). The binding of both cyclohexene nucleosides in the active site of HSV-1 thymidine kinase was investigated, and a model for the binding of both enantiomers is proposed. The amino acids involved in binding of the optical antipodes are the same, but the interaction energy of both enantiomers is slightly different. This may be attributed to the interaction of the secondary hydroxyl function of the nucleoside analogues with Glu-225. Structural analysis has demonstrated the flexibility of the cyclohexenyl system, and this may be considered as an important conformational characteristic explaining the potent antiviral activity. PMID- 10691699 TI - Acyclic analogues of deoxyadenosine 3',5'-bisphosphates as P2Y(1) receptor antagonists. AB - P2Y(1) receptors are activated by ADP and occur on endothelial cells, smooth muscle, epithelial cells, lungs, pancreas, platelets, and in the central nervous system. With the aid of molecular modeling, we have designed nucleotide analogues that act as selective antagonists at this subtype. The present study has tested the hypothesis that acyclic modifications of the ribose ring, proven highly successful for nucleoside antiviral agents such as gancyclovir, are generalizable to P2Y receptor ligands. Specifically, the binding site of the P2Y(1) receptor was found to be sufficiently accommodating to allow the substitution of the ribose group with acyclic aliphatic and aromatic chains attached to the 9 position of adenine. Three groups of adenine derivatives having diverse side chain structures, each containing two symmetrical phosphate or phosphonate groups, were prepared. Biological activity was demonstrated by the ability of the acyclic derivatives to act as agonists or antagonists in the stimulation of phospholipase C in turkey erythrocyte membranes. An acyclic N(6)-methyladenine derivative, 2-[2-(6-methylamino-purin-9-yl)-ethyl]-propane-1, 3 bisoxy(diammoniumphosphate) (10), containing an isopentyl bisphosphate moiety, was a full antagonist at the P2Y(1) receptor with an IC(50) value of 1.60 micro?. The corresponding 2-Cl derivative (11) was even more potent with an IC(50) value of 0.84 microM. Homologation of the ethylene group at the 9-position to 3-5 methylene units or inclusion of cis- or trans-olefinic groups greatly reduced antagonist potency at the P2Y(1) receptor. Analogues containing a diethanolamine amide group and an aryl di(methylphosphonate) were both less potent than 10 as antagonists, with IC(50) values of 14 and 16 microM, respectively, and no agonist activity was observed for these analogues. Thus, the ribose moiety is clearly not essential for recognition by the turkey P2Y(1) receptor, although a cyclic structure appears to be important for receptor activation, and the acyclic approach to the design of P2 receptor antagonists is valid. PMID- 10691700 TI - Conjugated enynes as nonaromatic catechol bioisosteres: synthesis, binding experiments, and computational studies of novel dopamine receptor agonists recognizing preferentially the D(3) subtype. AB - To evaluate nonaromatic catechol bioisosteres, the conformationally restrained enynes 1 and enediynes 2 were synthesized via palladium-catalyzed coupling as the key reaction step. Subsequent receptor binding studies at the dopamine receptor subtypes D(1), D(2 long), D(2 short), D(3), and D(4) showed highly interesting binding profiles for the enynes 1a and 1b when compared to dopamine. At the guanine nucleotide-sensitive high-affinity binding site of the D(3) receptor, the target compound 1b (K(i) = 5.2 nM) was 10-fold more potent than dopamine but less potent at the D(2) and D(4) subtypes. In contrast to dopamine the agonists 1a and 1b showed strong selectivity for the receptors of the D(2) family (D(2)-D(4)). As far as we know, this study represents the first report on nonaromatic dopamine agonists. Comparison of molecular electrostatic potentials, derived from semiempirical molecular orbital calculations, and lipophilicity maps was performed. PMID- 10691701 TI - Identification of deoxymiroestrol as the actual rejuvenating principle of "Kwao Keur", Pueraria mirifica. The known miroestrol may be an artifact. AB - Miroestrol (1) has been isolated previously as an active principle from "Kwao Keur" (Pueraria mirifica), a rejuvenating folk medicine from Thailand. Reinvestigation using bioassay-guided purification has resulted in the isolation of a new potent phytoestrogen, deoxymiroestrol (2). The facile aerial oxidation of 2 into 1 suggests the possibility that 1 may be an artifact. PMID- 10691702 TI - New circulin macrocyclic polypeptides from Chassalia parvifolia. AB - Four new macrocyclic polypeptides were isolated and identified from an extract of the tropical tree Chassalia parvifolia. Circulins C-F are 29-30 amino acid cyclic peptides in which the entire primary amino acid chain is covalently cyclized via peptide bonds. Their structures were deduced from a combination of FABMS analyses, N-terminal Edman degradation, endoproteinase digestion, and amino acid analyses. All the peptides share a high degree of sequence homology and contain six cysteine residues forming three intramolecular disulfide bridges. Circulins C F inhibited the cytopathic effects of in vitro HIV-1 infection with EC(50) values of 50-275 nM. PMID- 10691703 TI - Rearrangement of O-cinnamoyltaxicin I to a novel C-13 spiro-taxane. AB - During the large-scale synthesis of an O-cinnamoyltaxicin I acetonide, an intermediate for the semisynthesis of 7-deoxypaclitaxel derivatives, side-product 3 was formed via a vinylogous retro-aldol reaction and a long-range hydride shift from O-cinnamoyltaxicin I (1) under alkaline reaction conditions. Compound 3 has two hemi-acetal bridges at C-1,C-9 and C-10,C-13. Compound 4 was formed from side product 3 under acidic reaction conditions and is the first C-13 spiro-taxane described in the literature. This spiro-taxane has two acetal bridges between C 1, C-13 and C-10,C-13. PMID- 10691704 TI - Secobotrytriendiol and related sesquiterpenoids: new phytotoxic metabolites from Botrytis cinerea. AB - Six new sesquiterpenoid metabolites (1, 3-7) have been isolated from Botrytis cinerea. Their structures were elucidated by means of MS and extensive NMR studies. The phytotoxic activities of these new products have been evaluated. PMID- 10691705 TI - Biosynthesis of 4'-O-methylpyridoxine (Ginkgotoxin) from primary precursors. AB - Cell suspension cultures of Ginkgo biloba and Albizia tanganyicensis were investigated for the presence of 4'-O-methylpyridoxine (ginkgotoxin, 2), the 4'-O methyl derivative of vitamin B(6) (pyridoxine, 1). The cultures produced the toxin even in the absence of vitamin B(6) (a common additive to plant cell culture media). This indicates that the pyridoxine ring system of ginkgotoxin is synthesized de novo by the cultured cells. A feeding experiment with D-[U (13)C(6)]glucose revealed that the mode of incorporation of label into the pyridoxine moiety of 2 matched that observed for 1 in Escherichia coli. Thus, the data obtained in this investigation provide independent proof supporting the current hypothesis on vitamin B(6) biosynthesis. The 4'-O-methyl group of ginkgotoxin (2) was labeled from L-[methyl-(13)C(1)]methionine. This indicates that ginkgotoxin is likely to be derived by 4'-O-methylation of pyridoxine (1). The G. biloba cell suspension culture may be a suitable system to get further insight into vitamin B(6) and/or ginkgotoxin biosynthesis. PMID- 10691706 TI - Quinolizidine alkaloids from Sophora alopecuroides. AB - A new matrine-type alkaloid, 7alpha-hydroxysophoramine (1), was isolated from the aerial parts of Sophora alopecuroides together with eight known alkaloids, 12beta hydroxysophocarpine (2), sophoramine (3), 14beta-hydroxymatrine, matrine, sophoridine, sophocarpine, adenocarpine, and baptifoline. The structures of compounds 1-3 were confirmed through single-crystal X-ray diffraction analysis. PMID- 10691707 TI - Novel polycyclic guanidine alkaloids from two marine sponges of the genus Monanchora. AB - Two marine sponges of the genus Monanchora (Poecilosclerida, Crambeidae) have been found to contain new polycyclic guanidine alkaloids bearing the (5,6,8b) triazaperhydroacenaphthylene skeleton. Their structures have been determined by detailed spectroscopic analysis. Dehydrobatzelladine C (1) has been isolated from M. arbuscula and crambescidins 359 (2) and 431 (3) from M. unguiculata. The chemotaxonomic implications of these findings are discussed. PMID- 10691708 TI - Yanucamides A and B, two new depsipeptides from an assemblage of the marine cyanobacteria Lyngbya majuscula and Schizothrix species. AB - Yanucamides A (1) and B (2) were isolated from the lipid extract of a Lyngbya majuscula and Schizothrixsp. assemblage collected at Yanuca Island, Fiji. The structures of compounds 1 and 2 were determined by spectroscopic methods. Both compounds contain a unique 2,2-dimethyl-3-hydroxy-7-octynoic acid, which has previously been described only as a component of kulolide-1 (3) and kulokainalide 1 (4), metabolites from the marine mollusk Philinopsis speciosa. Thus, the isolation of the yanucamides from this cyanobacterial assemblage supports the hypothesis that the kulolides and related metabolites are of cyanobacterial origin. PMID- 10691709 TI - New quinolizidine alkaloids from Ulex jussiaei. AB - From an acidic extract of Ulex jussiaei four new quinolizidine alkaloids, jussiaeiines A (1), B (2), C (3), and D (4), were isolated and characterized by spectroscopic methods together with (-)-cytisine, (-)-N-methylcytisine, and (-) anagyrine. The proposed biosynthetic origin of these new alkaloids is briefly discussed. PMID- 10691710 TI - New isomalabaricane triterpenes from the marine sponge Stelletta globostellata that induce morphological changes in rat fibroblasts. AB - Three new isomalabaricane triterpenes, 29-hydroxystelliferin D (2), 3-epi-29 hydroxystelliferin E (3), and 3-epi-29-hydroxystelliferin A (4), were isolated from the marine sponge Stelletta globostellata. Their structures, including absolute stereochemistry, were determined on the basis of spectral data and chemical methods. Rat fibroblasts treated with 0.2 microM of 2-4 exhibited unusual morphological characteristics, followed by death in 5 days. PMID- 10691711 TI - New triterpenoid sulfates from the red alga Tricleocarpa fragilis. AB - Ten new sulfated terpenoids, including six cycloartenol sulfates (1-6), two 29 nor-cycloartenol sulfates (7,8), and two 29-nor-lanosterol sulfates (9,10), were isolated from brine shrimp-toxic fractions of the methanolic extract of the red alga Tricleocarpa fragilis collected in Hawaiian waters. Structures 1-10 were elucidated by spectral methods, and the absolute stereochemistry for compound 1 at C23 was determined by Mosher analysis. Compounds 7 and 10 showed brine shrimp toxicity at 50 microg/mL, while 1 and 3 showed substantial activity at 17 microg/mL. Compounds 2, 4, 5, and 9 were inactive. In cytotoxicity assays, compounds 1-10 were inactive at concentrations tested. PMID- 10691712 TI - Isolation and biochemical characterization of a new topoisomerase I inhibitor from Ocotea leucoxylon. AB - In a continuation of our search for potential tumor inhibitors from plants, we found that a crude extract from Ocotea leucoxylon showed selective activity typical of inhibitors of the enzyme topoisomerase I in a yeast assay for DNA damaging agents. Using a bioassay-directed fractionation approach, the major bioactive compound was isolated and identified as the known aporphine alkaloid dicentrinone (4); the inactive alkaloid dicentrine (3) was also isolated. Compound 4 showed selective bioactivity against the rad52 repair-deficient yeast strain RS322 (IC(12) 49 microg/mL) and was inactive against the rad52- and topo1 deficient strain RS321 (IC(12) > 2000 microg/mL) and against the repair proficient strain RJ03 (IC(12) > 2000 microg/mL). Biochemical studies with recombinant human topoisomerase I indicated that dicentrinone (4) is an inhibitor of the human enzyme. Colony formation studies suggest that it is weakly cytotoxic, but that its mechanism of toxicity differs from that of camptothecin and its derivatives. PMID- 10691713 TI - Three new diterpenoids based on the novel sarcopetalane skeleton from Croton sarcopetalus. AB - The roots of Croton sarcopetalus afforded three new diterpenoids (2-4) with a novel carbon skeleton that seems to be derived biosynthetically from a pimarane precursor. The essential oil of the roots gave trans-methylisoeugenol as the main constituent, along with 22 further compounds. PMID- 10691714 TI - Grindelane diterpenoids from Stevia subpubescens. AB - Four new 9R,13R-epoxylabdane diterpenes (1-4) and a known clerodane derivative, 3,4beta-epoxy-5beta,10beta-cis-17alpha, 20alpha-clerod-13(14)-en-15,16-olide, were isolated from the leaves of Stevia subpubescens. The structures, which correspond to the grindelane class of diterpenoids, were elucidated by NMR data, chemical correlation, and single-crystal X-ray diffraction analysis of monoacetate 5. The absolute configuration of 1-4 is based on the optical activity of ketone 3 as compared with data of closely related substances. PMID- 10691715 TI - Ferrocene-based electroactive derivatizing reagents for the rapid selective screening of alcohols and phenols in natural product mixtures using electrospray tandem mass spectrometry. AB - The alcohols and phenols of oil of cloves, lemon oil, rose absolut, and oil of peppermint were derivatized with ferroceneoyl azide to generate their ferroceneoyl carbamates. These derivatives are selectively detected at the attomole level, in nanomolar concentrations by electrospray-tandem mass spectrometry (ES-MS/MS) without the need for sample cleanup. The ES-MS/MS analyses of the four essential oils revealed all the expected alcohols, and, in the case of lemon oil, it detected alpha-terpineol as a trace component that was not readily observed by GC-MS. The ES-MS/MS analyses complements the more conventional GC-MS analysis. The ES-MS method has the advantage of speed, selectivity, and sensitivity over GC-MS for detection of a targeted alcohol of a specific mass or structural type. The ES-MS method does not require a chromatographic separation of the components to accomplish its task. In contrast, GC-MS remains the preferred method for the determination of the total constituents of an oil. The ES-MS method may produce artifact ions, especially if the sample is wet and an excess of the ferroceneoyl azide is used; however, the artifacts did not interfere with the analyses. PMID- 10691716 TI - Inhibitory effects on HIV-1 protease of constituents from the wood of Xanthoceras sorbifolia. AB - From a methanolic extract of the wood of Xanthoceras sorbifolia, two new compounds, 29-hydroxy-3-oxotirucalla-7,24-dien-21-oic acid (3, xanthocerasic acid) and epigallocatechin-(4beta-->8, 2beta-->O-7)-epicatechin (6), were isolated, together with 11 known compounds. Of the isolated compounds, 3 oxotirucalla-7, 24-dien-21-oic acid (2), oleanolic acid (4), and 6 were found to be inhibitory substances against human immunodeficiency virus (HIV-1) protease, with their 50% inhibitory concentrations (IC(50)) being 20, 10, and 70 microg/mL, respectively. Condensed tannins of high molecular weights with epicatechin and epiafzelechin as the main extender units were found to be the most active principles of this plant (IC(50) values ca. 6.0 microg/mL). PMID- 10691717 TI - Artoindonesianin C, a new xanthone derivative from Artocarpus teysmanii. AB - A new xanthone derivative, artoindonesianin C (1), was isolated from Artocarpus teysmanii, together with two known prenylated flavonoids, cycloartobiloxanthone and artonin J. The structure of artoindonesianin C (1) was determined on the basis of MS and NMR evidence and by comparison with known related compounds. PMID- 10691718 TI - Efficient synthesis of octandrenolone and related dipyranoacetophenones. AB - Octandrenolone (1) was prepared in high yield by condensation of 2', 4',6' trihydroxyacetophenone with 3-chloro-3-methylbut-1-yne in the presence of a catalytic amount of copper(I) iodide. Methylation of 1 afforded O methyloctandrenolone (2). Oxidation of 2 with m-chloroperoxybenzoic acid followed by hydrolysis gave the racemic trans-(+)-1-(9,10-dihydro-9,10-dihydroxy-5-methoxy 2,2,8, 8-tetramethyl-2H,8H-benzo[1,2-b:3,4-b']dipyran-6-yl)ethanone (3), which confirmed the structure of the natural product previously isolated from Melicope erromangensis. PMID- 10691719 TI - Retamatrioside, a new flavonol triglycoside from Retama sphaerocarpa. AB - A new flavonol triglycoside, retamatrioside (1), has been isolated from the aerial parts of Retama sphaerocarpa. The structure of 1 has been determined as rhamnazin 3-O-beta-D-glucopyranosyl-(1-->5)-[beta-D-apiofuranosyl(1-->2)]-al pha L-arabinofuranoside, using spectroscopic methods. PMID- 10691720 TI - Triterpenoid saponins from Bongardia chrysogonum. AB - Two new triterpenoid saponins, 3-O-[beta-D-glucopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->4)-alph a-L-arabinopyranosyl]-hederagenin (1) and 3-O-[beta-D glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-alph a-L-arabinopyranosyl] hederagenin 28-O-[beta-L-glucopyranosyl-(1-->6)-beta-L-glucopyranosyl] ester (2), together with five known saponins, were isolated from an ethanolic extract of the tubers of Bongardia chrysogonum. The structures of 1 and 2 were determined on the basis of spectroscopic studies. PMID- 10691721 TI - New xenicane diterpenoids from the gorgonian Acalycigorgia inermis. AB - Acalycixeniolides D-G [corrected] (1-4), four new diterpenoids of the xenicane class, have been isolated from the gorgonian Acalycigorgia inermis. The structures of these compounds have been determined by combined spectroscopic methods. These compounds exhibited cytotoxicity against a human leukemia cell line. PMID- 10691722 TI - A new bioactive triene aldehyde from the marine sponge Leucetta microraphis. AB - A new triene aldehyde, (2E,6Z,9Z)-2-methyl-2,6,9-icosatrienal (1), was isolated from a MeOH extract of the Okinawan marine sponge Leucetta microraphis. The structure of 1 was determined by spectroscopic analysis. Three imidazole alkaloids, leucettamine B, preclathridine A, and (9E)-clathridine 9-N-(2 sulfoethyl)imine, were also obtained and identified. Compound 1 showed moderate growth-inhibitory activity toward HeLa S3 cells. PMID- 10691723 TI - 1,9-Dimethylhypoxanthine from a southern Australian marine sponge spongosorites species. AB - A Spongosorites sp. collected off southern Australia has yielded 1, 9 dimethylhypoxanthine (4). The structure for 4 was solved by spectroscopic analysis. PMID- 10691724 TI - Chemistry of verongida sponges. 10. Secondary metabolite composition of the caribbean sponge Verongula gigantea. AB - A detailed analysis of the secondary metabolites of the Caribbean sponge Verongula gigantea has been performed. A number of bromotyrosine derivatives, 1, 2, and 6-17, were identified, one of which (17) is a novel compound. Its structure was determined on the basis of spectroscopic evidence. Additionally, aureol (18) and 5, 6-dibromo-N,N-dimethyltryptamine (19) were isolated from one of the five analyzed specimens. PMID- 10691725 TI - Nonpolar components of the latex of Euphorbia peplus. AB - The less polar fractions of the latex of Euphorbia peplus were found to contain obtusifoliol, cycloartenol, 24-methylenecycloartanol, lanosterol, and 24 methylenelanosterol in the free and esterified triterpene alcohol fractions; 9 cis-tricosene as the major component of the hydrocarbon fraction; and a new acyclic triterpene alcohol named peplusol (1). The structure of 1 was determined as the R-isomer of (all-E)-2-(5,9-dimethyl-1-methylene-4,8-decadienyl)-5,9, 13 trimethyl-4,8,12-tetradecatrien-1-ol by spectral and chemical methods. PMID- 10691726 TI - Structure and antibacterial activity ofp6 new labdane diterpenoid from Salvia leriaefolia. AB - Phytochemical investigation of the choloroform extract of Salvia leriaefolia afforded 8(17),12E,14-labdatrien-6,19-olide (1), and its structure was determined by a combination of spectral methods. Compound 1 was found to possess antibacterial activity against Staphylococcus aureus. PMID- 10691727 TI - Polyhalogenated monoterpenes from a tasmanian collection of the red seaweed Plocamium cartilagineum. AB - Two new polyhalogenated monoterpenes (3E, 7E)-8-bromo-(2E)-chloromethylene-(5R, 6R)-dichloro-6-methyloctadien-1-al (1) and (1Z,3E,7E)-8,9-dibromo-(1Z,5R*, 6R*,9) tetrachloro-6-methyloctatriene [corrected] (2), together with two known compounds (3 and 4), were isolated and identified from the red alga Plocamium cartilagineum collected along the eastern coast of Tasmania. The structures were established by spectroscopic techniques. PMID- 10691728 TI - Cernuosides A and B, two sucrase inhibitors from Pulsatilla cernua. AB - Two oleanane-type oligoglycosides, cernuosides A (1) and B (2), were isolated from the roots of Pulsatilla cernua (Ranunculaceae). Structure elucidation was accomplished by 1D and 2D NMR (DQF-COSY, TOCSY, HMQC, HMBC, and ROESY) methods, FABMS, and hydrolysis. Both compounds showed moderate activity against sucrase. PMID- 10691729 TI - Dolastatin 3 and two novel cyclic peptides from a palauan collection of Lyngbya majuscula. AB - A collection of Lyngbya majuscula from Palau contained the peptides dolastatin 3 (1), homodolastatin 3 (2), and kororamide (3), together with aplysiatoxin (4), debromoaplysiatoxin (5), and oscillatoxin A (6). The structures of the new peptides homodolastatin 3 (2) and kororamide (3) were determined by interpretation of spectroscopic data and chemical degradation. PMID- 10691730 TI - Isolation of the pharmacologically active saponin ginsenoside Rb1 from ginseng by immunoaffinity column chromatography. AB - Immunoaffinity column chromatography using an anti-ginsenoside Rb1 monoclonal antibody has made possible a single-step separation of ginsenoside Rb1 from a crude extract of ginseng roots (Panax ginseng). The combination of immunoaffinity column chromatography and an enzyme-linked immunosorbent assay (ELISA) was also investigated. PMID- 10691731 TI - Loss of PTEN facilitates HIF-1-mediated gene expression. AB - In glioblastoma-derived cell lines, PTEN does not significantly alter apoptotic sensitivity or cause complete inhibition of DNA synthesis. However, in these cell lines PTEN regulates hypoxia- and IGF-1-induced angiogenic gene expression by regulating Akt activation of HIF-1 activity. Restoration of wild-type PTEN to glioblastoma cell lines lacking functional PTEN ablates hypoxia and IGF-1 induction of HIF-1-regulated genes. In addition, Akt activation leads to HIF 1alpha stabilization, whereas PTEN attenuates hypoxia-mediated HIF-1alpha stabilization. We propose that loss of PTEN during malignant progression contributes to tumor expansion through the deregulation of Akt activity and HIF-1 regulated gene expression. PMID- 10691732 TI - ATR disruption leads to chromosomal fragmentation and early embryonic lethality. AB - Although a small decrease in survival and increase in tumor incidence was observed in ATR(+/-) mice, ATR(-/-) embryos die early in development, subsequent to the blastocyst stage and prior to 7.5 days p.c. In culture, ATR(-/-) blastocysts cells continue to cycle into mitosis for 2 days but subsequently fail to expand and die of caspase-dependent apoptosis. Importantly, caspase independent chromosome breaks are observed in ATR(-/-) cells prior to widespread apoptosis, implying that apoptosis is caused by a loss of genomic integrity. These data show that ATR is essential for early embryonic development and must function in processes other than regulation of p53. PMID- 10691734 TI - Translation by ribosome shunting on adenovirus and hsp70 mRNAs facilitated by complementarity to 18S rRNA. AB - Translation initiation on eukaryotic mRNAs involves 40S ribosome association with mRNA caps (m(7)GpppN), mediated by initiation factor eIF4F. 40S eukaryotic ribosomes and initiation factors undergo 5' scanning to the initiation codon, with no known role for complementarity between eukaryotic 18S rRNA and the 5' noncoding region of mRNAs. We demonstrate that the 5' noncoding region of human adenovirus late mRNAs, known as the tripartite leader, utilizes a striking complementarity to 18S rRNA to facilitate a novel form of translation initiation referred to as ribosome shunting, in which 40S ribosomes bind the cap and bypass large segments of the mRNA to reach the initiation codon. Related elements are also shown to promote ribosome shunting in adenovirus IVa2 intermediate phase mRNA during virus infection and in human heat shock protein 70 (hsp70) mRNA for selective translation during heat shock. The importance of mRNA complementarity to 18S rRNA suggests that ribosome shunting may involve either specific RNA structural features or a prokaryotic-like interaction between mRNA and rRNA. PMID- 10691733 TI - The carboxyl terminus of vertebrate poly(A) polymerase interacts with U2AF 65 to couple 3'-end processing and splicing. AB - Although it has been established that the processing factors involved in pre-mRNA splicing and 3'-end formation can influence each other positively, the molecular basis of this coupling interaction was not known. Stimulation of pre-mRNA splicing by an adjacent cis-linked cleavage and polyadenylation site in HeLa cell nuclear extract is shown to occur at an early step in splicing, the binding of U2AF 65 to the pyrimidine tract of the intron 3' splice site. The carboxyl terminus of poly(A) polymerase (PAP) previously has been implicated indirectly in the coupling process. We demonstrate that a fusion protein containing the 20 carboxy-terminal amino acids of PAP, when tethered downstream of an intron, increases splicing efficiency and, like the entire 3'-end formation machinery, stimulates U2AF 65 binding to the intron. The carboxy-terminal domain of PAP makes a direct and specific interaction with residues 17-47 of U2AF 65, implicating this interaction in the coupling of splicing and 3'-end formation. PMID- 10691735 TI - The p23 molecular chaperones act at a late step in intracellular receptor action to differentially affect ligand efficacies. AB - Multiple molecular chaperones, including Hsp90 and p23, interact with members of the intracellular receptor (IR) family. To investigate p23 function, we compared the effects of three p23 proteins on IR activities, yeast p23 (sba1p) and the two human p23 homologs, p23 and tsp23. We found that Sba1p was indistinguishable from human p23 in assays of seven IR activities in both animal cells and in yeast; in contrast, certain effects of tsp23 were specific to that homolog. Transcriptional activation by two IRs was increased by expression of any of the p23 species, whereas activation by five other IRs was decreased by Sba1p or p23, and unaffected by tsp23. p23 was expressed in all tissues examined except striated and cardiac muscle, whereas tsp23 accumulated in a complementary pattern; hence, p23 proteins might contribute to tissue-specific differences in IR activities. Unlike Hsp90, which acts on IR aporeceptors to stimulate ligand potency (i.e., hormone-binding affinity), p23 proteins acted on IR holoreceptors to alter ligand efficiencies (i.e., transcriptional activation activity). Moreover, the p23 effects developed slowly, requiring prolonged exposure to hormone. In vitro, p23 interacted preferentially with hormone-receptor-response element ternary complexes, and stimulated receptor-DNA dissociation. The dissociation was reversed by addition of a fragment of the GRIP1 coactivator, suggesting that the two reactions may be in competition in vivo. Our findings suggest that p23 functions at one or more late steps in IR-mediated signal transduction, perhaps including receptor recycling and/or reversal of the response. PMID- 10691736 TI - Homeodomain and winged-helix transcription factors recruit activated Smads to distinct promoter elements via a common Smad interaction motif. AB - We have investigated the regulation of the activin-inducible distal element (DE) of the Xenopus goosecoid promoter. The results show that paired-like homeodomain transcription factors of the Mix family, Mixer and Milk, but not Mix.1, mediate activin/TGF-beta-induced transcription through the DE by interacting with the effector domain of Smad2, thereby recruiting active Smad2/Smad4 complexes to the Mixer/Milk-binding site. We identify a short motif in the carboxyl termini of Mixer and Milk, which is demonstrated to be both necessary and sufficient for interaction with the effector domain of Smad2 and is required for mediating activin/TGF-beta-induced transcription. This motif is not confined to these homeodomain proteins, but is also present in the Smad2-interacting winged-helix proteins Xenopus Fast-1, human Fast-1, and mouse Fast-2. We demonstrate directly that transcription factors of different DNA-binding specificity recruit activated Smads to distinct promoter elements via a common mechanism. These observations, together with the temporal and spatial expression patterns of Mixer and Milk, lead us to propose a model for mesoendoderm formation in Xenopus in which these homeodomain transcription factor/Smad complexes play a role in initiating and maintaining transcription of target genes in response to endogenous activin-like signals. PMID- 10691737 TI - Yeast heterochromatin is a dynamic structure that requires silencers continuously. AB - Transcriptional silencing of the HM loci in yeast requires cis-acting elements, termed silencers, that function during S-phase passage to establish the silent state. To study the role of the regulatory elements in maintenance of repression, site-specific recombination was used to uncouple preassembled silent chromatin fragments from silencers. DNA rings excised from HMR were initially silent but ultimately reactivated, even in G(1)- or G(2)/M-arrested cells. In contrast, DNA rings bearing HML-derived sequence were stably repressed due to the presence of a protosilencing element. These data show that silencers (or protosilencers) are required continuously for maintenance of silent chromatin. Reactivation of unstably repressed rings was blocked by overexpression of silencing proteins Sir3p and Sir4p, and chromatin immunoprecipitation studies showed that overexpressed Sir3p was incorporated into silent chromatin. Importantly, the protein was incorporated even when expressed outside of S phase, during G(1) arrest. That silencing factors can associate with and stabilize preassembled silent chromatin in non-S-phase cells demonstrates that heterochromatin in yeast is dynamic. PMID- 10691738 TI - Mammalian hepatocyte differentiation requires the transcription factor HNF 4alpha. AB - HNF-4alpha is a transcription factor of the nuclear hormone receptor family that is expressed in the hepatic diverticulum at the onset of liver development. Mouse embryos lacking HNF-4alpha fail to complete gastrulation due to dysfunction of the visceral endoderm. This early embryonic lethality has so far prevented any analyses of the contribution of HNF-4alpha toward liver development and hepatocyte differentiation. However, we have shown that complementation of HNF 4alpha(-/-) embryos with a tetraploid embryo-derived wild-type visceral endoderm rescues this early developmental arrest and allows HNF-4alpha(-/-) embryos to proceed normally through midgestation stages of development. Examination of these rescued embryos revealed that HNF-4alpha was dispensable for specification and early development of the liver. However, HNF-4alpha(-/-) fetal livers failed to express a large array of genes whose expression in differentiated hepatocytes is essential for a functional hepatic parenchyma, including genes encoding several apolipoproteins, metabolic proteins, and serum factors. In addition, we have demonstrated that HNF-4alpha is essential for expression of the transcription factors HNF-1alpha and PXR within the fetal liver. We therefore conclude that HNF 4alpha is both essential for hepatocyte differentiation during mammalian liver development and also crucial for metabolic regulation and liver function. PMID- 10691739 TI - A Kruppel-like zinc finger protein is involved in nitrogen-fixing root nodule organogenesis. AB - Mechanisms regulating plant host differentiation of the nitrogen-fixing root nodules remain mostly unknown. Sinorhizobium meliloti induces this process in Medicago sativa in which the Mszpt2-1 gene is expressed in vascular bundles of roots and nodules. This gene codes for a Kruppel-like zinc finger protein, a class of transcription factors involved in many animal developmental processes. Expression of Mszpt2-1 in yeast cells conferred osmotic tolerance. Antisense plants grew normally but developed nonfunctional nodules, in which differentiation of the nitrogen-fixing zone and bacterial invasion were arrested. Hence, a vascular bundle-associated Kruppel-like gene is required for the formation of the central nitrogen-fixing zone of the root nodule. PMID- 10691740 TI - The stringent response in Myxococcus xanthus is regulated by SocE and the CsgA C signaling protein. AB - Myxococcus xanthus fruiting body development is induced by amino acid limitation. The decision to grow or develop is established by the RelA-dependent stringent response and A-signaling. We identified two new members of this regulatory hierarchy, socE and the C-signaling gene csgA. SocE depletion arrests growth and induces sporulation under conditions that normally favor growth as well as curtailing DNA and stable RNA synthesis, inhibiting cell elongation, and inducing accumulations of the stringent nucleotides ppGpp and pppGpp [(p)ppGpp]. This system separates C-signaling, which does not occur under these conditions, from CsgA enzyme activity. Amino acid substitutions in the CsgA coenzyme binding pocket or catalytic site eliminate growth arrest. relA mutation also eliminates growth arrest. Eleven pseudorevertants selected for growth following SocE depletion contained mutations in csgA or relA. These results suggest that CsgA induces the stringent response and while SocE inhibits it. Unlike the csgA mutant, wild-type and socE csgA cells maintained high levels of (p)ppGpp throughout development. We suggest that CsgA maintains growth arrest throughout development to divert carbon from A-signaling and other sources into developmental macromolecular synthesis. PMID- 10691742 TI - A hard road: finding ways to reduce teen tobacco use. PMID- 10691741 TI - Progression of meiotic DNA replication is modulated by interchromosomal interaction proteins, negatively by Spo11p and positively by Rec8p. AB - Spo11p is a key mediator of interhomolog interactions during meiosis. Deletion of the SPO11 gene decreases the length of S phase by approximately 25%. Rec8p is a key coordinator of meiotic interhomolog and intersister interactions. Deletion of the REC8 gene increases S-phase length, by approximately 10% in wild-type and approximately 30% in a spo11Delta background. Thus, the progression of DNA replication is modulated by interchromosomal interaction proteins. The spo11 Y135F DSB (double strand break) catalysis-defective mutant is normal for S-phase modulation and DSB-independent homolog pairing but is defective for later events, formation of DSBs, and synaptonemal complexes. Thus, earlier and later functions of Spo11 are defined. We propose that meiotic S-phase progression is linked directly to development of specific chromosomal features required for meiotic interhomolog interactions and that this feedback process is built upon a more fundamental mechanism, common to all cell types, by which S-phase progression is coupled to development of nascent intersister connections and/or related aspects of chromosome morphogenesis. Roles for Rec8 and/or Spo11 in progression through other stages of meiosis are also revealed. PMID- 10691743 TI - From social taboo to "torch of freedom": the marketing of cigarettes to women. PMID- 10691744 TI - Asia: choppy seas for BAT butt boat. PMID- 10691745 TI - India: movie shoots at women. PMID- 10691747 TI - Australia: philip morris exploits SIDS research PMID- 10691746 TI - Czech Republic: Gauloises dates for students. PMID- 10691748 TI - Litigation: a tobacco lawyer's view PMID- 10691749 TI - South africa: one life, waste it! PMID- 10691751 TI - UK: no cigarette under the stiff upper lip PMID- 10691752 TI - Holy smoke! PMID- 10691750 TI - It's true. It kills. It's great! PMID- 10691753 TI - Hong kong: down at the fair. PMID- 10691754 TI - Smoke signs: patterns of tobacco billboard advertising in a metropolitan region. AB - OBJECTIVE: To use geographic information systems data and analyses to describe locations and characteristics of tobacco billboards in a large metropolitan area, and to assess the extent to which tobacco companies are locating billboards in close proximity to minority neighbourhoods and schools. DESIGN: Observational study of billboards in a large metropolitan region. SETTING: City and county of St Louis, Missouri. PARTICIPANTS: All stationary billboards in the city and county of St Louis were eligible to be observed, with the exception of bus stop and street side retail advertising signs (for example, cigarette advertising at gas stations). A total of 1239 non-blank billboards were observed. All data were collected in early 1998. MAIN OUTCOME MEASURES: Tobacco and non-tobacco billboard geographic distribution; billboard type and product brand frequencies; and billboard neighbourhood characteristics. RESULTS: Almost 20% of the billboards contained tobacco advertising. Four of the top five and nine of the top 22 brands displayed on billboards were tobacco products. Billboards were located in all areas of St Louis except for the communities with the highest average incomes. Tobacco billboards were more likely to be found in low income areas and areas with a higher percentage of African Americans. Images of African American figures on tobacco billboards were concentrated in the most heavily African American populated regions of the city. Approximately 74% of all billboards in the city of St Louis were within 2000 feet (700 metres) of public school property. CONCLUSIONS: Tobacco products were the single most heavily advertised type of product on billboards in St Louis. The geographic distribution of tobacco billboards, as well as the types of images found on these billboards, is consistent with the hypothesis that tobacco companies are targeting poor and minority communities with their advertising. Methods employing geographic information systems are a powerful technique for examining outdoor tobacco advertising. PMID- 10691755 TI - A randomised controlled trial of a community intervention to prevent adolescent tobacco use. AB - OBJECTIVE: Experimental evaluation of comprehensive community wide programme to prevent adolescent tobacco use. DESIGN: Eight pairs of small Oregon communities (population 1700 to 13 500) were randomly assigned to receive a school based prevention programme or the school based programme plus a community programme. Effects were assessed through five annual surveys (time 1-5) of seventh and ninth grade (ages 12-15 years) students. INTERVENTION: The community programme included: (a) media advocacy, (b) youth anti-tobacco activities, (c) family communications about tobacco use, and (d) reduction of youth access to tobacco. MAIN OUTCOME MEASURE: The prevalence of self reported smoking and smokeless tobacco use in the week before assessment. RESULTS: The community programme had significant effects on the prevalence of weekly cigarette use at times 2 and 5 and the effect approached significance at time 4. An effect on the slope of prevalence across time points was evident only when time 2 data points were eliminated from the analysis. The intervention affected the prevalence of smokeless tobacco among grade 9 boys at time 2. There were also significant effects on the slope of alcohol use among ninth graders and the quadratic slope of marijuana for all students. CONCLUSION: The results suggest that comprehensive community wide interventions can improve on the preventive effect of school based tobacco prevention programmes and that effective tobacco prevention may prevent other substance use. PMID- 10691756 TI - Competence skills help deter smoking among inner city adolescents. AB - OBJECTIVE: To test whether higher levels of general competence are linked to more frequent use of refusal assertiveness that is in turn related to less subsequent smoking among inner city adolescents. METHODS: Longitudinal study conducted during three year middle school or junior high school period. A sample of 1459 students attending 22 middle (ages 11-14 years) and junior high (ages 12-15 years) schools in New York City participated. Students completed surveys at baseline, one year follow up, and two year follow up. The students self reported smoking, decision making skills, personal efficacy, and refusal assertiveness. Teams of three to five data collectors administered the questionnaire following a standardised protocol. These data were collected in school during a regular 40 minute class period. RESULTS: Based on the tested structural equation model, decision making and personal efficacy (that is, general competence) predicted higher refusal assertiveness and this greater assertiveness predicted less smoking at the two year follow up. The tested model had a good fit and was parsimonious and consistent with theory. CONCLUSIONS: Adolescent smoking prevention programmes often teach refusal skills in order to help youth resist peer pressure to smoke. The present findings suggest that teaching general competence skills as well may help to reduce smoking because youth with better personal efficacy and decision making skills are better able to implement smoking refusal strategies. PMID- 10691757 TI - Effect of an eight week smoking ban on women at US navy recruit training command. AB - OBJECTIVE: To examine the effect of a unique organisational smoking ban on female United States Navy recruits, a population with historically high smoking rates. SETTING AND DESIGN: Study participants were female recruits (n = 5503) entering the Navy recruit training command between March 1996 and March 1997 (12 consecutive months). Participants completed smoking surveys at entry to recruit training (baseline) and again at graduation from training after exposure to an eight week, 24 hour a day smoking ban. Effects of the ban on baseline to graduation changes in perceptions of being a smoker were examined, and relapse rates among baseline ever smokers was assessed three months after leaving recruit training. RESULTS: Among all recruits, 41.4% reported being smokers at entry (that is, reported any smoking in the 30 days before entering recruit training). As a result of the ban, there was a significant reduction (from about 41% to 25%, p < 0.001) in the percentage of all women recruits who reported themselves as smokers, a much larger change than expected had no ban been in place. Relapse at the three month follow up varied according to the type of smoker at entry into the Navy, with rates ranging from 89% relapse among baseline daily smokers to 31% among baseline experimenters. CONCLUSIONS: Findings suggest that the ban provides some smokers who desire to quit with an external impetus and support to do so. However, high relapse rates indicate that more than an organisationally mandated smoking ban during recruit training is needed to help younger smokers, more regular smokers, and those who intend to continue smoking to quit after joining the Navy. PMID- 10691759 TI - Exposure of black youths to cigarette advertising in magazines. AB - OBJECTIVE: To estimate the potential exposure of black adolescents to brand specific advertising in magazines. DESIGN: A probit regression analysis was conducted of pooled 1990 and 1994 data on brand specific advertising in 36 popular US magazines to examine the relationship between the presence or absence of advertising in each magazine for each of 12 cigarette brands, and the proportion of each magazine's youth (ages 12-17 years) readers who were black. MAIN OUTCOME MEASURES: The presence or absence of advertising in each magazine in 1990 and 1994, for each of 12 cigarette brands. RESULTS: After controlling for total magazine readership and the percentage of young adult, Hispanic, and female readers, black youth cigarette brands (those whose market share among black youths exceeded their overall market share) were more likely than other brands to advertise in magazines with a higher percentage of black youth readers. Holding all other variables constant at their sample means, the probability of a non black youth brand advertising in a magazine decreased over the observed range of percentage black youth readership from 0.65 (95% confidence interval (CI) 0.55 to 0.75) for magazines with 5% black youth readers to 0.33 (95% CI 0.00 to 0.69) for magazines with 91% black youth readers. In contrast, the probability of a black youth brand advertising in a magazine increased from 0.40 (95% CI 0.17 to 0.62) at 5% black youth readership to 1.00 (95% CI 0.97 to 1.00) at 91% black youth readership. CONCLUSIONS: Black youths are more likely than white youths to be exposed to magazine advertising by cigarette brands popular among black adolescents. PMID- 10691758 TI - Investing in youth tobacco control: a review of smoking prevention and control strategies. AB - OBJECTIVE: To provide a comprehensive review of interventions and policies aimed at reducing youth cigarette smoking in the United States, including strategies that have undergone evaluation and emerging innovations that have not yet been assessed for efficacy. DATA SOURCES: Medline literature searches, books, reports, electronic list servers, and interviews with tobacco control advocates. DATA SYNTHESIS: Interventions and policy approaches that have been assessed or evaluated were categorised using a typology with seven categories (school based, community interventions, mass media/public education, advertising restrictions, youth access restrictions, tobacco excise taxes, and direct restrictions on smoking). Novel and largely untested interventions were described using nine categories. CONCLUSIONS: Youth smoking prevention and control efforts have had mixed results. However, this review suggests a number of prevention strategies that are promising, especially if conducted in a coordinated way to take advantage of potential synergies across interventions. Several types of strategies warrant additional attention and evaluation, including aggressive media campaigns, teen smoking cessation programmes, social environment changes, community interventions, and increasing cigarette prices. A significant proportion of the resources obtained from the recent settlement between 46 US states and the tobacco industry should be devoted to expanding, improving and evaluating "youth centred" tobacco prevention and control activities. PMID- 10691760 TI - Case study of attempts to enact self service tobacco display ordinances: a tale of three communities. AB - OBJECTIVE: To examine self service tobacco displays (SSTDs) and youth retail tobacco access by comparing longitudinal illegal tobacco sales rates in three communities in Santa Barbara County, California, that considered or implemented ordinances banning SSTDs. A confirmatory survey was also conducted to substantiate the longitudinal data. DESIGN: A longitudinal case study design was utilised. Five undercover tobacco buys were conducted between 1994 and 1997 (n = 332). In addition, one confirmatory survey was conducted in a geographically separated area, which had no ordinances banning SSTDs (n = 57). RESULTS: Decreases in youth buy rates were reported in all three communities. Most notably, the first city to enact a SSTD ban, Carpinteria, achieved a 0% sales rate, which was maintained throughout the study period. In contrast, Santa Barbara and Goleta experienced considerable drops in their illegal sales rates, but neither community obtained results as dramatic as those found in Carpinteria. The confirmatory survey showed that 32.1% of stores with SSTDs sold cigarettes to minors; this compares to a sales rate of 3.4% in stores without SSTDs (chi(2) (1) = 8.11, p = 0.004). CONCLUSIONS: Efforts to enact self service bans are likely to meet with retail and tobacco industry opposition, as was the case in this study's three communities. The process of community debate, resultant publicity surrounding the issue, and enactment of SSTD ordinances may serve to not only increase merchant awareness of youth tobacco laws and their penalties but also may contribute to reduced youth cigarette sales rates. Implications and limitations of the findings are discussed. PMID- 10691761 TI - The economics of tobacco: myths and realities. PMID- 10691762 TI - Reflections on the saga of tar content: why did we measure the wrong thing? PMID- 10691763 TI - Banning smoking outdoors is seldom ethically justifiable. PMID- 10691764 TI - Banning outdoor smoking is scientifically justifiable. PMID- 10691765 TI - Outdoor smoking bans: more than meets the eye. PMID- 10691766 TI - How I nearly became a Marlboro man. PMID- 10691767 TI - Abreast of the West: German effort to distract poles from the truth about smoking. PMID- 10691769 TI - Tobacco and women's health PMID- 10691768 TI - Only naughty the first time. PMID- 10691771 TI - Smoke and mirrors: a history of denial PMID- 10691770 TI - Double indemnity: making sense of the US settlement PMID- 10691772 TI - Up from the ashes: the fight for a new tobacco Act PMID- 10691774 TI - Introduction to the Fourth Acta Physiologica Scandinavica International Symposium on vasodilators in the development of hypertension: NO and dopamine. PMID- 10691773 TI - Catecholamine biosynthesis and physiological regulation in neuroendocrine cells. AB - The catecholamines are widely distributed in mammals and their levels and physiological functions are regulated at many sites. These include their release from neuroendocrine cells, the type and sensitivity of the multiple receptors in target cells, the efficacy of the reuptake system in the secretory cells, and the rates of catecholamine biosynthesis and degradation. In the present review the main focus will be on the more recent studies on the biosynthesis in neuroendocrine cells which involves a specific set of enzymes, with special reference to physiologically important regulatory mechanisms. Eight enzymes of the biosynthetic pathway have now been identified, cloned, expressed as recombinant proteins, characterized with respect to catalytic and regulatory properties, and some of them also crystallized. The identification of the tyrosine hydroxylase catalysed reaction as the rate-limiting step in the normal catecholamine biosynthesis has attracted most attention, both in terms of transcriptional and post-translational regulation. In certain human genetic disorders of catecholamine biosynthesis other enzymes in the pathway may become rate-limiting, notably those involved in the biosynthesis/regeneration of the natural co-factor tetrahydrobiopterin in the tyrosine hydroxylase reaction. The enzymes involved seem to be regulated by a variety of physiological factors, both on a long-term scale and a short-term basis, and include the relative rates of synthesis, degradation and state of activation of the biosynthetic enzymes, notably of tyrosine hydroxylase. Multiple surface receptors and signalling pathways are activated in response to extracellular stimuli and play an essential role in the regulation of catecholamine biosynthesis. PMID- 10691775 TI - Biological effects of arginine metabolites. AB - Arginine and its metabolites exert physiological effects on the vasculature and on the kidney and also provide important influences on the regulation of cell proliferation. We summarize the known information regarding two major metabolites of arginine: (a) nitric oxide (NO) and (b) agmatine, decarboxylated arginine. Both agents appear to interact in producing vasodilation and increases in glomerular filtration rate (GFR) in the kidney. There is evidence for inter regulation of arginine pathways in the sense that agmatine is capable of inhibiting inducible nitric oxide synthase (iNOS), the inflammatory NOS isoform. Both NO and agmatine influence cell proliferation via effects on polyamine synthesis. In addition, both NO and agmatine exert inhibitory effects on ornithine decarboxylase (ODC) and the putrescine transporter by significantly different mechanisms. Therefore, arginine and arginine metabolites exert both vascular regulatory functions and impact on the regulation of cell proliferation. Significant inter-regulation among arginine pathways occurs within the three metabolic major pathways within the cell: (1) nitric oxide synthase (2) arginase and ornithine decarboxylase, and (3) arginine decarboxylase. PMID- 10691776 TI - Protein-protein interactions controlling nitric oxide synthases. AB - Nitric oxide (NO) biosynthesis is tightly regulated by a variety of mechanisms ranging from transcriptional to post-translational controls. Calmodulin has long been known to be an allosteric modulator of the three major NO synthases (NOS). Recent studies indicate that other proteins directly associate with NOS isoforms and regulate their activity or spatial distribution in the cell. Several proteins residing in or recruited to plasmalemmal caveolae of endothelial cells serve as allosteric regulators of endothelial NOS (eNOS). Caveolins, the resident scaffolding proteins of caveolae, and calmodulin undergo reciprocal Ca2+ dependent association and dissociation with eNOS in the caveolar membrane that inhibits (caveolins) and activates (calmodulin) eNOS activity. Other caveolar proteins appear to contribute to the eNOS-membrane complex, including the bradykinin B2 receptor, the angiotensin AT1 receptor, the CAT1 arginine transporter, and Hsp90. Direct interactions of a variety of proteins bearing PDZ domains with the PDZ domain of neuronal NOS (nNOS) have been shown to influence the subcellular distribution and/or activity of the enzyme in brain and muscle. One of these proteins, PSD-93, co-localizes with a subpopulation of nNOS in the macula densa. Although considerable emphasis has been placed on transcription as the principal step of regulation for inducible NOS (iNOS), our laboratory has recently defined a regulatory interaction of iNOS with Rho family GTPases. While the role of protein-eNOS interactions in the control of vascular tone has been increasingly clarified, the interactions and regulatory importance of protein association with nNOS and iNOS in the vasculature and kidney remains to be explored. PMID- 10691777 TI - A pivotal role of nitric oxide in endothelial cell dysfunction. AB - The functional role of the vascular endothelium is a subject of growing interest and appreciation. Some of the key functions of the endothelium are modulated by the activity and expression of endothelial nitric oxide synthase (eNOS), suggesting a role for this enzyme in endothelial dysfunction. Several well-known angiogenic stimulators exert their effect only in the presence of the functional eNOS. In this setting NO production is responsible for the scalar podokinetic cell motility, which is a prerequisite for the acquisition of vectorial movement when guidance cues are applied. The mode of this NO action appears to lie in the accelerated turnover of focal adhesions through the process of activation/inactivation of protein tyrosine phosphatases. Localization of eNOS to the caveolar domains, in the proximity of clustered beta1 integrins, provides an additional level of regulatory complexity through the modulation of caveolar dynamics and the state of caveolin oligomerization. Therefore, eNOS serves various important functions in the endothelium and is a putative target for therapeutic interventions. PMID- 10691778 TI - Mode of nitric oxide action on the renal vasculature. AB - Our study aimed to characterize the essential cellular pathways along which nitric oxide (NO) exerts its well-known vasodilatatory properties in the kidney. Using the isolated perfused rat kidney model we examined the roles of potassium channels, cGMP-protein kinase activity and cAMP-phosphodiesterases (PDE) in the effect of NO on renovascular resistance. We found that neither potassium channel activity nor G-kinase activity was essential for the vasodilatatory effect of NO. The effect of NO, however, was essentially mimicked by pharmacological inhibition of PDE-3, which is a cGMP-inhibitable PDE. As PDE-3 is strongly expressed in renal preglomerular vessels and NO stimulates cGMP formation in renal vessels, it appears likely that inhibition of cAMP degradation and consequently the cAMP pathway are crucially involved in mediating the effects of NO on renal vascular resistance. PMID- 10691779 TI - Interactions of the renin-angiotensin system and neuronal nitric oxide synthase in regulation of cyclooxygenase-2 in the macula densa. AB - Cyclooxygenase-2 (COX-2) expression in rat kidney is localized to the macula densa and the immediately proximal cTALH and increases after salt restriction. Either ACE inhibitors or AT1 receptor blockers increase COX-2 expression in both control and salt-restricted animals, suggesting that the RAS activation feedback inhibits renal cortical COX-2 expression. To determine whether increased COX-2 expression in response to ACE inhibition mediated increases in renin production, rats were treated with Captopril for 1 week with or without the specific COX-2 inhibitor, SC58236. Plasma renin activity increased significantly in the Captopril group. This increase was partially reversed by simultaneous treatment with SC58236. Kidney renin activity also increased in the Captopril group compared with control, which was also significantly inhibited by SC58236 treatment. Because of the localization of bNOS to MD and surrounding cTALH, the current study investigated the role of NO in the regulation of COX-2 expression. Rats were fed a normal diet, low salt diet or low salt diet combined with captopril and half of them were treated with the neuronal NOS inhibitor, 7-NI, and half with vehicle. After 7 days, mRNA was extracted and the microsome proteins purified from renal cortex. COX-2 mRNA expression was measured by Northern-blot and normalized with GAPDH. 7-NI treatment decreased COX-2 mRNA and immunoreactive COX-2 expression in each group. In summary, these studies indicate that COX-2 from macula densa/cTALH is a regulator of renin production and release. Angiotensin II may be a negative regulator of cTALH/macula densa COX-2 expression, and NO may mediate increased renal cortical COX-2 expression seen in volume depletion. These studies suggest important interactions between the NO and COX-2 systems in the regulation of arteriolar tone and the renin-angiotensin system by the macula densa. PMID- 10691780 TI - The autoinhibitory control element and calmodulin conspire to provide physiological modulation of endothelial and neuronal nitric oxide synthase activity. AB - NO production by the endothelial and neuronal isoforms of nitric oxide synthase (cNOS) is regulated on a moment-to-moment basis by calmodulin binding, triggered by transient elevations in intracellular-free calcium levels. Nonetheless, additional modes of cNOS regulation are implicit in the discoveries of stimuli that elicit a sustained increase in cNOS activity despite undetectable or transient increases in intracellular Ca2+ in endothelial cells; such stimuli include shear-stress, oestrogen, insulin or insulin-like growth factor treatment of endothelial cells. Recently, we identified a peptide insertion within the FMN binding domain of mammalian NOSs that is unique to calcium-dependent isoforms, and not shared with inducible NOS or ancestral flavoproteins. Evidence suggests that this insertion serves as a fundamental control element, analogous to intrinsic autoinhibitory peptides that have been demonstrated to regulate activity of other calmodulin-dependent enzymes. Thus, the peptide insertion of cNOSs appears to function as structural element that is displaced upon calmodulin binding, resulting in dysinhibition of NO synthesis. Once displaced, the peptide may also be subject to transient chemical modifications and protein-protein interactions that modulate autoinhibitory function. Herein we summarize our present knowledge and speculate on mechanisms by which calmodulin and the autoinhibitory peptide conspire to regulate cNOS activity. PMID- 10691781 TI - Nitric oxide: a physiological mediator of the type 2 (AT2) angiotensin receptor. AB - Virtually all of the biological actions of angiotensin II (ANG II) have been thought to be mediated by the type 1 (AT1) angiotensin receptor and the function of the type 2 (AT2) receptor is unknown. We now describe a novel physiological action of ANG II to release nitric oxide (NO) mediated by the AT2 receptor in both the kidney and gastrointestinal tract. We present an integrated model for a counter-regulatory protective action of the AT2 receptor mediated by nitric oxide. In the kidney, ANG II at the AT2 receptor stimulates a vasodilator cascade of bradykinin (BK), NO and cyclic GMP which is tonically activated only during conditions of increased ANG II, such as sodium depletion. In the absence of the AT2 receptor, pressor and antinatriuretic hypersensitivity to ANG II is associated with BK and NO deficiency. In angiotensin-dependent hypertension, the hypotensive effect at AT1 receptor blockade is due at least in part to AT2 receptor stimulation and consequent increased activity of the vasodilator cascade. In the gastrointestinal tract, physiological quantities of ANG II stimulate the AT2 receptor releasing NO and cGMP leading to increased sodium and water absorption. In conclusion, NO is an important physiological mediator of ANG II at the AT2 receptor. PMID- 10691782 TI - Intracellular localization of dimethylarginine dimethylaminohydrolase overexpressed in an endothelial cell line. AB - Methylarginines are endogenous inhibitors of nitric oxide synthase (NOS) and have been implicated in the regulation of the nitric oxide pathway in health and disease. Cellular concentrations of free methylarginines are determined in part by the activity of dimethylarginine dimethylaminohydrolase (DDAH). There are two isoforms of DDAH which have distinct tissue distributions with some relationship to NOS isoforms. We have determined the intracellular localization of both DDAH isoforms by overexpression of epitope-tagged DDAH in an immortalized endothelial cell line. Immunofluorescence confocal microscopy and immunoblotting indicate that both isoforms are predominantly cytosolic with no specific association with organelles or the plasma membrane. These data suggest that the key role for DDAH may be to ensure that under normal conditions the levels of methylarginines are kept low throughout the whole cell. PMID- 10691783 TI - Phosphorylation and activation of the endothelial nitric oxide synthase by fluid shear stress. AB - Fluid shear stress activates the endothelial nitric oxide (NO) synthase (eNOS) by a mechanism which does not require an increase in the intracellular concentration of free Ca2+ ([Ca2+]i), and is sensitive to several kinase inhibitors. Although phosphorylation of eNOS has been suggested to regulate enzyme activity, the mechanism of eNOS activation is still unclear. Here we demonstrate that fluid shear stress elicits the phosphorylation of eNOS on tyrosine and serine residues. Inhibition of phosphatidylinositol 3-kinase (PI3K), using wortmannin or a dominant negative mutant of its downstream target, Akt (protein kinase B), prevented the maintained serine phosphorylation and activation of eNOS. Enhancing eNOS phosphorylation by inhibiting serine/threonine phosphatases, increased eNOS activity by approximately twofold, as assessed by the accumulation of intracellular cyclic GMP, without increasing the intracellular concentration of free Ca2+. These data suggest that shear stress activates a pathway involving PI3K and the serine/threonine kinase Akt, which phosphorylates eNOS. This phosphorylation directly increases eNOS activity at resting [Ca2+]i, thus rendering the shear stress-induced activation of eNOS apparently Ca2+ independent. PMID- 10691785 TI - Exocytosis and endocytosis in juxtaglomerular cells. AB - The cellular events related to secretion of renin are not well understood. Here we review some of the evidence that has led to the current understanding of renin secretion as a process that involves exocytosis as the predominant mode of secretion. This is based on the observation of occasional fusion events between secretory granules and cell membrane and measurement of intermittent secretion of renin from single afferent arterioles, with a renin content of each secretion episode that corresponds to the renin content of one secretory granule. More recently it has been demonstrated that the afferent arterioles lose a large number of renin granules after acute stimulation without changing the average granular volume. Current electrophysiological techniques have now permitted direct measurements of cell membrane capacitance in juxtaglomerular (JG) cells as a measure of net addition (exocytosis) or removal (endocytosis) of membrane material. With this technique we have shown that cAMP, which is a vasodilator and stimulates renin secretion, enhances net exocytosis at low concentrations, while at higher concentrations membrane retrieval processes are also stimulated. We suggest that both exocytosis and endocytosis are regulated processes in the JG cells and both may be important for the long-term control of renin secretion at the single cell level. PMID- 10691784 TI - Impaired effect by NO synthase inhibition on tubuloglomerular feedback in rats after chronic renal denervation. AB - Acute unilateral renal denervation (aDNX) is associated with reduced tubuloglomerular feedback (TGF) sensitivity. Six days after denervation (cDNX) TGF sensitivity is somewhat restored, but TGF reactivity increased. This study aimed to investigate if the increased TGF reactivity that was seen in cDNX kidneys was owing to reduced production of nitric oxide (NO). TGF characteristics were determined with micropuncture experiments in anaesthetized rats, using the stop-flow pressure (PSF) technique. Maximal drop in PSF (DeltaPSF) was used as an index of TGF reactivity and the loop of Henle perfusion rate that elicited half maximal DeltaPSF, the turning point (TP) was used as a measure of TGF sensitivity. In cDNX kidneys, TP was higher than in control rats (25.4 +/- 1.5 nL min-1 vs. 19.1 +/- 1.1 nL min-1), but clearly lower than in aDNX rats (37. 3 +/- 3.1 nL min-1). TGF was more reactive in cDNX rats (DeltaPSF=14. 7 +/- 1.1 mmHg) than in aDNX (7.9 +/- 1.1 mmHg) and control rats (9. 6 +/- 0.9 mmHg). Intratubular inhibition of NO synthase N omega-nitro-L-arginine (L-NA) in sham DNX animals, decreased TP to 13.9 +/- 2.2 nL min-1 and DeltaPSF was increased with 92%. In cDNX kidneys TP was not significantly reduced by L-NA, and TGF reactivity was only moderately increased by 31%. Intratubular infusion of L arginine (L-Arg) reduced DeltaPSF from 10.2 +/- 0.7 to 6.5 +/- 0.6 mmHg in sham DNX kidneys, but TP was unaffected. In cDNX kidneys, there was no effect on either DeltaPSF or TP by the addition of L-Arg. However, when NO was delivered via sodium nitroprusside in the tubular perfusate, a clear reduction of DeltaPSF was seen in both sham-DNX and cDNX kidneys (from 9.9 +/- 0.5 to 4.4 +/- 1.0 and from14.9 +/- 1.3 to 8.1 +/- 1.5 mmHg, respectively). This indicates that cDNX is a state of low renal NO production and that this low level of NO resets TGF to a higher sensitivity and more pronounced reactivity. PMID- 10691786 TI - Advanced glycosylation end-products and NO-dependent vasodilation in renal afferent arterioles from diabetic rats. AB - Systemic pressor responses to acetylcholine (ACh) are reduced in DM, an effect thought to be related to quenching of nitric oxide (NO) by advanced glycosylation end-products (AGE). We studied the effects of AGE in juxtamedullary (JM) afferent arterioles (AA) from rats with 40-50 days diabetes mellitus (DM) induced via streptozotocin. JM AA were perfused in vitro with solutions containing fresh RBCs suspended in either 6% bovine albumin or 6% AGE-albumin in euglycaemic Krebs Ringer. Autoregulatory responses were evident in the DM vessels: AA constricted 31 +/- 2% (n=9) when perfusion pressure (PP) was raised from 60 to 140 mmHg. ACh (10 microM) caused a 43 +/- 15% dilation and Ca2+-channel blockade elicited a 95 +/- 14% dilation at 100 mmHg PP, indicating substantial basal vascular tone in DM AA. L-NAME (0.1 mM) constricted DM AA by 21 +/- 2% (n=9) at 100 mmHg PP, indicating significant basal NO production in DM vessels. Segments of renal resistance arteries from DM rats perfused in vitro responded to muscarinic stimulation and elevated glucose levels with significant increments in NO production, as measured with an NO-sensitive electrode. This observation shows that the renal endothelial NO system is intact in DM. While AGE in the perfusate dilated control AA, they had no effect on DM AA at all PP levels, although they blunted ACh-induced dilation. Hence, although AGE do appear to have vasoactive properties in the absence of hyperglycaemia, the results of this study are inconsistent with substantial NO quenching by AGE. PMID- 10691787 TI - Endogenous nitric oxide and epoxyeicosatrienoic acids modulate angiotensin II induced constriction in the rabbit afferent arteriole. AB - Nitric oxide (NO) and epoxyeicosatrienoic acids (EETs), cytochrome P450 epoxygenase metabolites of arachidonic acid, are released by the vascular endothelium and play important roles in the control of glomerular haemodynamics. We examined whether endogenous NO or EETs modulate angiotensin II- (AngII) induced constriction in isolated microperfused afferent arteriole (Af-Art) of the rabbit kidney. When Af-Arts were treated with NG-nitro-L-arginine methyl ester (L NAME, an inhibitor of NO synthese; 10-4 mol L-1) or miconazole (an inhibitor of P450 epoxygenase; 10-6 mol L-1), basal diameter was decreased by 34.5 +/- 2.2 and 13.9 +/- 3.2%, respectively. AngII added to both the bath and lumen decreased the diameter of Af-Arts in a dose-dependent manner. Pretreatment with either L-NAME or miconazole also augmented the constrictor response to AngII. AngII at 10-8 mol L-1 decreased the diameter to 39.2 +/- 1.4, 32.9 +/- 3.6, and 12.7 +/- 4.6%, in control, L-NAME-, and miconazole-treated group, respectively. In order to study whether the AngII type2 (AT2) receptor modulates AngII action via NO or EETs, we repeated the experiments in the presence of PD123319 (an AT2 receptor antagonist; 10-7 mol L-1). In the presence of PD123319, L-NAME still augmented the constrictor response to AngII, however, miconazole had no effect. In the presence of PD123319, AngII at 10-8 mol L-1 decreased the diameter to 25.0 +/- 4.6, 9.4 +/ 4.0, and 26.0 +/- 3.3%, in control, L-NAME-, and miconazole-treated group, respectively. These results suggest that (1) tonic release of NO and EETs attenuates the vasoconstrictor response to AngII in Af-Arts and (2) AT2 receptor seems to be coupled to EETs rather than the NO pathway. PMID- 10691788 TI - Chronic increases in transmural pressure reduce NO-mediated dilations in isolated resistance arteries of the hamster. AB - It is unclear whether the impairment of NO-mediated dilation in hypertension is the cause or the consequence of high blood pressure. We therefore studied in isolated resistance arteries whether elevated transmural pressure affects NO mediated dilation. Arteries (n=5-7) were perfused at hydrostatic pressures of either 45, 120 or 160 mmHg for 48 h. Subsequently, diameter and calcium responses (fura 2) were studied at a transmural pressure of 45 mmHg. Pre-exposure to 120 and 160 mmHg reduced resting diameters and minimal diameters after stimulation with noradrenaline and significantly increased corresponding intracellular free calcium levels in vascular smooth muscle. Moreover, the NO-mediated dilation in response to acetylcholine was significantly reduced although the increase in endothelial calcium was not altered. Dilations induced by the NO donor SNP were not affected. It is concluded that chronically elevated pressure per se impairs endothelial NO production by a mechanism distal to receptor-dependent calcium increases. PMID- 10691789 TI - Interaction between nitric oxide and oxygen radicals in regulation of tubuloglomerular feedback. AB - NADPH oxidase, nitric oxide synthase (NOS) and cyclooxygenase are oxidases that are expressed in the juxtaglomerular apparatus (JGA) or blood vessels and can generate oxygen radicals (O-2) during partial reduction of molecular oxygen. O-2 interacts rapidly and irreversibly with nitric oxide (NO) to yield peroxynitrite (ONOO-), thereby restricting the half-life, diffusion distance and bioactivity of NO in tissues. NO generated by a neuronal (n) NOS isoform that is heavily expressed in macula densa (MD) cells, is generated during NaCl reabsorption at the MD and blunts the expression of the tubuloglomerular feedback (TGF) response. Therefore, we tested the hypothesis that O-2 formed in the JGA of the normal rat limits NO signalling. Tempol is a membrane-permeable superoxide dismutase (SOD) mimetic. Maximal TGF responses were assessed from the fall in proximal stop flow pressure during orthograde perfusion of artificial tubular fluid (ATF) into the loop of Henle. Microperfusion of tempol (10-4 M) into the efferent arteriole (EA) of Wistar-Kyoto rats blunted maximal TGF response (8. 2 +/- 0.4 vs. 6.4 +/- 0.4 mmHg; n=8; P < 0.05). Graded doses of the NO donor compound, S-nitroso acetylpenicillamine (SNAP; 10-7-10-4 M) microperfused into the lumen of the MD produces graded buffering of TGF. During EA microperfusion of tempol, responses to luminal SNAP at 10-6 M and greater were enhanced significantly (P < 0.05 or <0. 01). In conclusion, O-2 generated in the JGA can be metabolized by a membrane permeable SOD mimetic. O-2 enhances the basal TGF response and limits NO signalling from the macula densa. Therefore, O-2 and NO interact in the JGA to modulate the TGF response. PMID- 10691790 TI - Concerted actions of renal endothelial and macula densa NO systems in the maintenance of extracellular fluid volume. AB - It is now clear that nitric oxide (NO) exerts a substantial influence on renal function and that the kidney has a high capacity to produce NO. However, there are at least two different NO systems in the kidney. The interplay between NO generated by the endothelium and by the macula densa is considered in this review. It seems that endothelial NO increases in response to an increase in perfusion pressure and an increase in distal delivery, whereas macula densa NO decreases upon a sustained increase in distal delivery. Furthermore, evidence is accumulating that macula densa NO may well mediate renin release. Though seemingly in contrast, both the response of the endothelial NO and of the macula densa NO system seem appropriate to restore a perturbation of fluid balance. The function of the tubuloglomerular feedback (TGF) mechanism is likely to be influenced by both sources of NO, because of the close proximity of these NO producing cells to the vascular smooth muscle cells of the afferent arteriole. The endothelial NO system seems to be responsible for short-term, dampening actions to increased afferent arteriolar tone elicited by activation of the TGF system. The macula densa NO system, on the other hand, is probably adapting TGF responses to sustained increases in distal delivery. The analysis presented in this paper is an attempt to integrate the function of the two NO systems into physiological regulation. The exact role of the medullary NOS enzymes remains to be further elucidated. PMID- 10691791 TI - Nitric oxide and preglomerular vascular lesions in lyon spontaneously hypertensive rats. AB - Accumulation of Sudan black-stainable (SB+) lipids is a hallmark of the focal inflammato-proliferative lesions that develop along preglomerular vessels in N G nitro-L-arginine methyl ester (L-NAME) and angiotensin II hypertensive rats. We extended our findings to genetically hypertensive Lyon (LH) rats aged 14 and 30 weeks and to age-matched normotensive (LN) rats. Vessels were isolated by HCl maceration. Despite high systolic blood pressure (SBP), hypercholesterolaemia, albuminuria and increased interlobular and afferent arteriolar media thickness, SB+ lesions were rarely found in LH rats, regardless of age. To probe nitric oxide as a potential source of vascular protection, 14-week-old LN and LH rats received L-NAME for 10 days (20 mg kg-1 day-1, per os), which increased SBP to 174 +/- 5 and to >200 mmHg, respectively. It induced formation of focal SB+ lesions less frequently in LN than LH rats, in which they affected 39 +/- 7, 44 +/- 5 and 15 +/- 5% of arcuate arterial branches, interlobular arteries and afferent arterioles, respectively. Immunoreactive endothelin-1 was found to accumulate at the level of SB+ lesions. Co-administered with L-NAME, hydralazine (15 mg kg-1 day-1, per os) limited SBP rise to approximately 10 mmHg in both LN and LH rats. As a result, SB+ lesions were rare in LN rats, but were frequent in LH rats. In conclusion, preglomerular SB+ lesions are spontaneously lacking in LH rats. Endogenous nitric oxide production provides protection against vascular barotrauma. Endothelin-1 likely plays an autocrine/paracrine role in vascular lesion formation. PMID- 10691792 TI - Nitric oxide-angiotensin II interactions in angiotensin II-dependent hypertension. AB - Many studies indicate that renal haemodynamic function in angiotensin II- (ANG II) dependent hypertension is not reduced as much as would be predicted from the elevated ANG II levels suggesting that counteracting renoprotective mechanisms are activated. One important renoprotective effect is mediated by increased levels of nitric oxide. Recent studies using the ANG II-infused hypertensive rat model have shown that inhibition of nitric oxide synthesis (NOS) causes greater decreases in renal blood flow and glomerular filtration rate in ANG II-infused hypertensive rats than in control rats. This augmented nitric oxide-dependent influence is localized primarily in the cortex and to the preglomerular vasculature. The differential effects on the renal cortex and medulla are also reflected by the differences in NOS activities and protein expression. Ca2+ dependent NOS activity was significantly greater in the cortex but not the medulla of the ANG II-infused hypertensive rats compared with control rats. This was associated with marked activation of endothelial NOS protein levels and smaller increases in neuronal NOS protein levels in the cortex but not in the medulla. In contrast, the Ca2+-independent NOS activity and the inducible NOS protein levels in the cortex were significantly lower in the ANG II-infused hypertensive rats. These data support the hypothesis that cortical Ca2+-dependent NOS, primarily endothelial NOS, is stimulated during the early phases of ANG II induced hypertension and exerts a renoprotective effect on cortical haemodynamics. PMID- 10691793 TI - Control of arterial blood pressure and renal sodium excretion by nitric oxide synthase in the renal medulla. AB - Work in our laboratory has focused on the role of nitric oxide synthase (NOS) in the regulation of renal medullary function. Biochemical studies demonstrated that the renal medulla is enriched in immunoreactive NOS protein and NOS enzymatic activity when compared with the renal cortex. Further experiments showed large amounts of NOS activity in the inner medullary collecting ducts, while moderate NOS activity was found in glomeruli and vasa recta and minimal NOS activity was detected in other nephron segments examined. In subsequent functional studies, selective renal medullary infusion of NOS stimulators (bradykinin or acetylcholine) or inhibitors (L-NAME) preferentially altered medullary blood flow. The alterations in medullary flow were associated with parallel changes in sodium and water excretion. Similar to the effects observed in anaesthetized rats, chronic infusion of L-NAME directly into the renal medullary interstitial space of conscious, uninephrectomized SD rats selectively decreased renal medullary blood flow throughout a 5-day L-NAME infusion period. The decrease in medullary blood flow was associated with retention of sodium and the development of hypertension, and the effects were reversible. In contrast to the effects of chronic NOS inhibition, renal medullary infusion of NOS substrate L-arginine prevented the development of sodium-sensitive hypertension in the Dahl salt sensitive rat placed on a high sodium diet. The data reviewed in this paper indicate that NOS isoforms expressed in the renal medulla have a potent influence on renal medullary tubular and vascular function with consequential effects on fluid and electrolyte homeostasis and arterial blood pressure. PMID- 10691794 TI - Blood pressure control in eNOS knock-out mice: comparison with other species under NO blockade. AB - Changes in arterial blood pressure (ABP) lead to changes in vascular shear stress. This mechanical stimulus increases cytosolic Ca2+ in endothelial cells, which in turn activates the endothelial isoform of the nitric oxide synthase. The subsequently formed NO reaches the adjacent vascular smooth muscle cells, where it reduces vascular resistance in order to maintain ABP at its initial level. Thus, NO may play an important role as a physiological blood pressure buffer. Previous data on the importance of eNOS for blood pressure control are reviewed with special emphasis on the fact that endogenous nitric oxide can buffer blood pressure variability (BPV) in dogs, rats and mice. In previous studies where all isoforms of the nitric oxide synthase were blocked pharmacologically, increases in blood pressure and variability were observed. Thus, we set out to clarify which isoform of the nitric oxide synthase is responsible for this BPV controlling effect. Hence, blood pressure control was studied in knock-out mice lacking specifically the gene for endothelial nitric oxide synthase with their respective wild-type controls. One day after surgery, under resting conditions, blood pressure was increased by 47 mmHg (P < 0.05), heart rate was lower (-77 beats min-1, P < 0.05), and BPV doubled (P < 0.05). Based on these results, we conclude that chronic blood pressure levels are influenced by eNOS and that there is a blood pressure buffering effect of endogenous nitric oxide which is mediated by the endothelial isoform of the nitric oxide synthase. PMID- 10691795 TI - Abnormal pressure-natriuresis in hypertension: role of nitric oxide. AB - The kidneys have a critical role in long-term control of arterial pressure by regulating extracellular fluid and plasma volume. According to the renal body fluid feedback mechanism for long-term control, persistent hypertension can only occur as a result of a reduction in renal sodium excretory function or a hypertensive shift in the pressure-natriuresis relationship. Although an abnormal relationship between renal perfusion pressure and renal sodium excretion has been identified in every type of hypertension where it has been sought, factors responsible for this effect are still unclear. Nitric oxide (NO) is produced within the kidney and plays an important role in the control of many intrarenal processes which regulate the renal response to changes in perfusion pressure and thus, help determine plasma volume and blood pressure. Numerous studies have shown that long-term inhibition of NO synthesis results in a chronic rightward shift and marked attenuation in renal pressure-natriuresis. Recent studies have shown that certain animal models of genetic hypertension and forms of human hypertension areas are associated with a decrease in NO synthesis. Reductions in NO synthesis reduces renal sodium excretory function not only through direct actions on the renal vasculature, but through modulation of other vasoconstrictor processes and through direct and indirect alterations in tubular sodium transport. The causes and consequences of the dysregulation of NO in hypertension and other renal disease processes remain an important area of investigation. PMID- 10691796 TI - Renal NO production and the development of hypertension. AB - The juxtaglomerular apparatus (JGA) has the very important functions of detecting the fluid flow rate to the distal tubule and thus controlling the glomerular filtration rate (GFR) (tubuloglomerular feedback mechanism [TGF]) and renin release from the afferent arteriole. In studies of the TGF it has been evident that the sensitivity of this mechanism can be reset. Volume expansion will reset it to a low sensitivity leading to a high GFR and urine excretion rate, while dehydration will sensitize the TGF mechanism, giving rise to a low GFR and low urine excretion rate. Furthermore, we have found that in animals that spontaneously develop hypertension there is initially a sensitization of the TGF, leading to a reduced GFR and urine excretion rate, with fluid volume retention in the body and a consequent rise in blood pressure. When the pressure is raised, the TGF characteristics are normalized. In the macula densa (MD) cells in the JGA, there is a large production of NO from neuronal NOS. This production continuously reduces TGF sensitivity and is apparently impaired in animals that spontaneously develop hypertension. When we added an nNOS inhibitor to the drinking water for several weeks while measuring blood pressure, we found an increase in blood pressure after 3-4 weeks of treatment. This effect was abolished by a high salt diet. From these investigations, it also appeared as if nNOS-derived NO inhibited renin release. Experiments have also indicated that NO may resensitize inhibited G-protein coupled purinergic receptors. PMID- 10691797 TI - Plasma from ESRD patients inhibits nitric oxide synthase activity in cultured human and bovine endothelial cells. AB - Our recent observations of reduced total nitric oxide synthesis in renal failure patients on peritoneal dialysis and haemodialysis suggest that hypertension in end-stage renal disease involves lack of vasodilatory endothelial NO. To directly test this, uraemic plasma was obtained from dialysis patients and incubated with cultured vascular endothelial cells, to determine the effect on nitric oxide synthase (NOS) activity in comparison with plasma from subjects with normal renal function. After incubation for 6 h with 20% uraemic plasma from peritoneal dialysis and immediately prehaemodialysis patients, NOS activity was reduced in human dermal microvascular endothelial cells. Haemodialysis did not remove the NOS-inhibitory activity of uraemic plasma nor did it activate inducible NOS, as NOS activity was always similar in control and dexamethasone pretreated cells. Nitric oxide production (accumulation of nitrite and nitrate) was lower in cells incubated with uraemic vs. normal plasma and excess arginine increased nitric oxide production by cells previously exposed to uraemic medium. This inhibitory effect was not associated with co-factor deficiency but did correlate with plasma concentrations of endogenous NOS inhibitors. These in vitro findings suggest that low endothelial NOS activity may contribute to hypertension in end stage renal disease patients. PMID- 10691798 TI - Role of nitric oxide in haemodialysis hypotension. AB - To investigate the possible involvement of nitric oxide (NO) in haemodialysis hypotension, we measured plasma concentrations of nitrate anion (NO3-), a metabolite of NO, in 114 patients undergoing maintenance haemodialysis. Mean plasma NO3- concentrations before dialysis were greater in subjects with lower blood pressure (155 +/- 16 micromol L-1) than in those with middle (117 +/- 8 micromol L-1) or higher blood pressure (105 +/- 12 micromol L-1) before dialysis. Further, mean plasma NO3- concentrations before dialysis were greater in subjects with lower blood pressure (186 +/- 13 micromol L-1) than in those with middle (112 +/- 7 micromol L-1) or higher blood pressure (64 +/- 11 micromol L-1) after dialysis. Plasma NO3- concentrations before dialysis were inversely correlated with mean blood pressure before dialysis (r=0.318, P=0.0006), and showed a strong inverse correlation with mean blood pressure after dialysis (r=0.608, P=0.0001). In the selected participants who had equal range of mean blood pressure before dialysis, mean plasma NO3- concentrations were greater in subjects with severe hypotension during dialysis (180 +/- 14 micromol L-1) than in those with mild hypotension (99 +/- 11 micromol L-1) or without hypotension (53 +/- 12 micromol L 1); plasma NO3- concentrations before dialysis were inversely correlated with changes in mean blood pressure during dialysis and mean blood pressure after dialysis. Results indicate that enhanced NO production may be involved in acute hypotension during dialysis, and suggest the possible involvement of NO in the pathogenesis of chronic hypotension associated with maintenance haemodialysis. PMID- 10691799 TI - Genetic mechanisms underlying the regulation of urinary sodium excretion and arterial blood pressure: the role of adducin. PMID- 10691800 TI - Differentiation of vasoactive renal sympathetic nerve fibres. AB - Activation of renal sympathetic nerves produces marked changes in renal haemodynamics, tubular ion and water transport and renin secretion. This review examines information indicating that these effects are mediated by functionally specific groups of renal sympathetic nerve fibres separately innervating the renal vessels, tubules and juxtaglomerular granular cells. PMID- 10691801 TI - Vasoconstrictor responses in thromboxane receptor knockout mice: tubuloglomerular feedback and ureteral obstruction. AB - The role of thromboxane (TP) in the vasoconstriction induced by tubuloglomerular feedback or 18-h ureteral obstruction was studied in wild type mice (TP +/+), and in heterozygous (TP +/-) and homozygous TP receptor knockout mice (TPR -/-). TGF function was assessed from the response of stop flow pressure (PSF) to a maximum increase in loop of Henle flow rate (0-30 nL min-1). PSF fell by 6.4 +/- 0.4 mmHg in wild-type mice, by 6.1 +/- 0.6 mmHg in TP +/-, and by 7.9 +/- 0.7 mmHg in TP /- mice. In the presence of the TP receptor agonist U46,619 (10-5 M) the PSF reduction increased to 10. 4 +/- 0.8 mmHg in TP +/+, and to 10.6 +/- 2.8 mmHg in TP +/-, but was unchanged at 7.7 +/- 0.7 mmHg in TP -/-. Mean arterial blood pressures were comparable between groups (103 +/- 3 mmHg in TP +/+, 113 +/- 4.6 in TP +/- and 113 +/- 2.4 mmHg in TP -/- mice). Intratubular pressure following unilateral ureteral obstruction was significantly higher in TP -/- than in TP +/+ mice both in the early phase (0-3 h) and late phase (18 h) of obstruction. These results indicate that chronic TP receptor deficiency does not significantly alter maximum TGF responses in mice, and that it is accompanied by exaggerated vasodilatation during short-term unilateral ureteral obstruction and attenuated vasoconstriction during longer lasting obstruction. We conclude that thromboxane is primarily a regulator of renal vascular tone under pathophysiological conditions. PMID- 10691802 TI - Renal sodium/calcium exchange; a vasodilator that is defective in salt-sensitive hypertension. AB - The Na+ : Ca2+ exchanger is an important plasma membrane ion transport pathway that plays a major role in controlling [Ca2+]i. In smooth muscle cells, it may function as a Ca2+ extrusion pathway and may help lower [Ca2+]i in response to vasoconstrictor-induced increases in [Ca2+]i. It may also extrude [Ca2+]i and lead to vasodilation in response to vasodilators. Our recent studies have been performed to determine the existence and regulation of the Na+ : Ca2+ exchanger in renal contractile cells which include afferent and efferent arterioles and mesangial cells. Exchanger activity is present in all three of these contractile elements but is higher in afferent arterioles vs. efferent arterioles. We have also examined the role of altered regulation of the exchanger in the SHR and in salt-sensitive hypertension. With the establishment of high blood pressure, Na+ : Ca2+ exchanger activity is reduced in afferent but not in efferent arterioles in both models of hypertension. Other works in cultured mesangial cells and freshly dissected afferent arterioles, have shown that protein kinase C (PKC) up regulates the Na+ : Ca2+ exchanger from Dahl/Rapp salt-resistant rats while it fails to do so in arterioles and mesangial cells from salt-sensitive rats. This defect in PKC regulation of Na+ : Ca2+ exchange is the result of a loss of PKC mediated translocation of the exchanger to the plasma membrane in S mesangial cells. Thus, a defect in the PKC-Na+ : Ca2+ exchanger-translocation pathway may cause dysregulation of [Ca2+]i and help explain the dramatic decrease in GFR that occurs in this model of hypertension. PMID- 10691803 TI - Intrarenal dopamine coordinates the effect of antinatriuretic and natriuretic factors. AB - The precision by which sodium balance is regulated suggests an intricate interaction between modulatory factors released from intra- and extrarenal sources. Intrarenally produced dopamine has a central role in this interactive network. Dopamine, produced in renal tubular cells acts as an autocrine and paracrine factor to inhibit the activity of Na+,K+-ATPase as well as of a number of sodium influx pathways. The natriuretic effect of dopamine is most prominent under high salt diet. The antinatriuretic effects of noradrenaline, acting on alpha-adrenoceptors and angiotensin II are opposed by dopamine as well as by atrial natriuretic peptide (ANP). Several lines of evidence have suggested that ANP acts via the renal dopamine system and recent studies from our laboratory have shown that this effect is attributed to recruitment of silent D1 receptors from the interior of the cell towards the plasma membrane. Taken together, the observations suggest that dopamine coordinates the effects of antinatriuretic and natriuretic factors and indicate that an intact renal dopamine system is of major importance for the maintenance of sodium homeostasis and normal blood pressure. PMID- 10691804 TI - Dopamine D(3) receptors in the rat kidney: role in physiology and pathophysiology. AB - It is well accepted that dopamine receptors play an important role in the regulation of cardiovascular and kidney function. Most of the knowledge on the renal actions of dopamine has been accumulated focussing on the prototypes of the two known dopamine receptor subfamilies, i.e. D1 and D2. The dopamine D3 receptor is a member of the D2-like subfamily and has been intensively studied in the neurosciences. Recently, the peripheral actions of this receptor subtype have also raised considerable interest as well because its effects on kidney function appear to be different from that of the other dopamine receptors. This short overview will summarize the data reported and add new results on the role of D3 receptors in the regulation of renal function as well as their potential pathophysiological implications. PMID- 10691805 TI - Effect of salt intake on jejunal dopamine, Na+,K+-ATPase activity and electrolyte transport. AB - The present study addresses the question of the relevance of salt intake on jejunal dopamine, Na+,K+-ATPase activity and electrolyte transport. Low salt, but not high salt, intake for 2 weeks increased dopamine levels in the jejunal mucosa accompanied by a marked decrease in L-3,4-dihydroxyphenylalanine tissue levels. By contrast, in rats fasted for 72 h the effect of refeeding for 24 h with a low salt diet failed to change dopamine tissue levels, although it significantly increased those of L-3,4-dihydroxyphenylalanine. By contrast, high salt intake markedly increased the tissue levels of both dopamine and L-3,4 dihydroxyphenylalanine, without changes in dopamine/L-3,4-dihydroxyphenylalanine tissue ratios. Tissue levels of both L-3,4-dihydroxyphenylalanine and dopamine in control conditions (normal salt intake for 2 weeks) were markedly higher (P < 0.05) than in rats submitted to 72 h fasting plus 24 h refeeding. The effect of fasting for 72 h followed by 24 h refeeding was a marked decrease in jejunal Na+,K+-ATPase activity, particularly evident for rats fed a normal salt and high salt diets during the refeeding period. Basal short circuit current was similar in rats fed a normal salt diet for 2 weeks and 24 h, and the type of diet failed to alter basal short circuit current after refeeding with normal, low and high salt diets. On the other hand, the effect of prolonged low salt intake was a marked decrease in jejunal Na+, K+-ATPase activity and basal short circuit current, whereas high salt intake failed to alter enzyme activity and basal short circuit current. In rats fed for 2 weeks a high salt diet ouabain was found to be more potent in reducing jejunal short circuit current than in rats fed normal and low salt diets. The effect of furosemide was more marked in rats fed for 2 weeks high and low salt diets than in animals receiving a normal salt intake. Dopamine (up to 1 micromol L-1) was found not to alter Na+,K+-ATPase and basal short circuit current in jejunal epithelial sheets, in rats fed with normal, low and high salt diets for 2 weeks and 24 h. PMID- 10691806 TI - Chemical hypoxia-induced increases in dopamine D1A receptor mRNA in renal epithelial cells are mediated by nitric oxide. AB - Nitric oxide (NO) and dopamine (DA) have similar effects on renal function, with both having natriuretic and diuretic effects mediated by vascular and tubular mechanisms. Renal ischaemia or hypoxia have been shown to influence the activity of both systems. However, it is not known whether there is any crosstalk between the NO and dopaminergic systems in the kidney. Here using the porcine proximal tubule-like renal epithelial LLC-PK1 cell line as a model system, we determined whether exposure of cells to chemical hypoxia altered the steady-state levels of D1A receptor mRNA and whether the changes involved the NO system. Exposure of LLC PK1 cells to chemical hypoxia resulted in a marked increase in D1A receptor mRNA levels as measured by reverse transcription-polymerase chain reaction (RT-PCR). The increased levels of D1A receptor mRNA following hypoxia were blocked by the NO synthase inhibitors NG-nitro-L-arginine methylester (L-NAME) or NG-monomethyl L-arginine (L-NMMA). Further evidence that the NO system exerted positive effects on D1A receptor gene expression came from finding that the NO donor sodium nitroprusside, the NO precursor L-arginine and the guanosine 3', 5'-cyclic monophosphate (cyclic GMP) analogue 8-Br-cGMP all increased D1A receptor mRNA levels in LLC-PK1 cells. These results indicate that expression of the D1A receptor in LLC-PK1 cells can be positively regulated by the NO system. Such an interaction between the renal NO and DA systems may contribute to the reported protective effects that NO and DA exert upon the kidney under conditions of ischaemia. PMID- 10691807 TI - The collecting duct, dopamine and vasopressin-dependent hypertension. AB - AVP not only increases osmotic water permeability (Pf) in the rat cortical collecting duct (CCD), but also acts synergistically with aldosterone to augment sodium reabsorption (JNa). These effects are inhibited by catecholamines via alpha2 adrenergic receptors, and by dopamine. We review here studies designed to determine the mechanism and receptor involved in dopamine action. The inhibitory effect of dopamine on Na+ and water transport was found to be reversible, and was not produced by agonists specific to D1A and D1B receptors. D2-type (D2, D3 or D4) receptors and activation of the GTP-binding protein Gi were implicated by the observation that dopamine had no inhibitory effect when JNa and Pf were stimulated by a cyclic AMP analogue plus isobutylmethylxanthine. The only dopaminergic antagonist that reversed the inhibitory effect of dopamine was clozapine, which is relatively D4-specific. We also found that dopamine or D1 specific agonists by themselves had no effect on cAMP production. However, dopamine inhibited the high rate of AVP-dependent cAMP production, and this effect of dopamine was reversed by clozapine but not other antagonists or by inhibitors of protein kinase C. The D4 receptor was observed in western blots of renal cortical proteins, and it was localized to the collecting duct by RT-PCR and immuno-histochemistry using a D4-specific antibody. These results show that at least a portion of the natriuretic effect of dopamine can be attributed to inhibition of AVP-dependent Na+ reabsorption by the CCD, and they introduce another signalling system as a candidate in the aetiology of low-renin, salt dependent hypertension. PMID- 10691808 TI - D1 dopamine receptor signalling defect in spontaneous hypertension. AB - Dopamine modulates cardiovascular function by actions in the central and peripheral nervous system, by altering the secretion/release of prolactin, pro opiomelanocortin, vasopressin, aldosterone, and renin, and by directly affecting renal function. Dopamine produced by the renal proximal tubule exerts an autocrine/paracrine action via two classes of dopamine receptors, D1-like (D1 and D5) and D2-like (D2, D3, and D4), that are differentially expressed along the nephron. The autocrine/paracrine function of dopamine, manifested by tubular rather than by haemodynamic mechanisms, becomes most evident during extracellular fluid volume expansion. This renal autocrine/paracrine function is lost in essential hypertension and in some animal models of genetic hypertension. The molecular basis for the dopaminergic dysfunction in hypertension may involve an abnormal post-translational modification of dopamine receptors. PMID- 10691809 TI - Defective renal dopamine D1-like receptor signal transduction in obese hypertensive rats. AB - It is reported that dopamine promotes renal sodium excretion via activation of D1 like dopamine receptors located on the proximal tubules. In spontaneously hypertensive rats the natriuretic and diuretic response to exogenously administered and endogenously produced dopamine is reduced, which results from a diminished dopamine-induced inhibition of the enzyme, Na+,K+-ATPase. The present study was designed to examine dopamine-receptor mediated inhibition of Na+,K+ ATPase and its associated signal transduction pathway in the proximal tubules of Zucker obese and lean control rats. The obese animals were hypertensive, hyperinsulinaemic and hyperglycaemic compared with the lean rats. While dopamine caused inhibition of Na+,K+-ATPase activity in lean rats, this effect was significantly attenuated in the obese animals. There was significant reduction in D1-like receptor numbers in the basolateral membranes of obese rats compared with lean rats with no change in the affinity to the ligand [3H]SCH 23390 between the two groups of rats. Dopamine failed to stimulate G proteins as measured by [35S]GTPgammaS binding in the obese rats. Also, dopamine was unable to cause phospholipase-C activation in obese rats, but it did activate phospholipase-C in lean rats. These results show that reduction in D1-like receptor numbers and a defect in receptor-G protein coupling may account for the inability of dopamine to activate the D1-like receptor-coupled signal transduction pathway and cause inhibition of Na+,K+-ATPase in the obese hypertensive rats. PMID- 10691810 TI - Renal dopamine and noradrenaline excretion during CNS-induced natriuresis in spontaneously hypertensive rats: influence of dietary sodium. AB - Abnormalities in dopamine (DA) and noradrenaline (NA) activities and sodium handling may be involved in the pathogenesis of hypertension. The present study was designed to investigate whether any differences exist between normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) in urinary excretion of DA, NA and sodium after 15 weeks on a low, medium or high sodium diet and during a subsequent elevation of the cerebroventricular fluid sodium concentration (CNS-induced natriuresis). Seven features were noted: (1) Basal sodium and DA excretion after the diet regimen was correlated to the dietary sodium content in both strains, except that sodium and DA excretion in SHR showed no further increase after the high sodium diet over and above that after medium sodium diet. (2) For any given sodium diet, SHR excreted more DA and NA as compared with WKY. (3) Blood pressure in SHR, as opposed to that in WKY, was higher after medium and high sodium diet than after low sodium diet. (4) During CNS-induced natriuresis NA excretion decreased or remained unchanged in WKY, but increased in SHR. (5) The DA/NA excretion ratio during CNS-induced natriuresis increased in WKY while decreased in SHR, which would not favour a natriuretic/vasodilatory response in the latter. (6) The ability of SHR to respond with CNS-induced natriuresis was attenuated after high sodium diet. (7) The magnitude of CNS-induced natriuresis was in both strains correlated to the sodium diet; the higher the dietary sodium content, the greater the natriuretic response. In conclusion, the study shows some clear differences in the catecholamine and sodium handling between WKY and SHR which may be involved in the pathogenesis of hypertension in SHR. Furthermore, increased sodium in the diet sensitizes the brain and kidney to increase the ability to respond with natriuresis for a given sodium stimulus. PMID- 10691811 TI - Do we do what they say we do? coding errors in urology. AB - OBJECTIVE: To determine the accuracy of routine data coding in a large multispeciality urological unit. Materials and methods From the clinical records, the diagnosis and procedure codes were ascribed to 106 finished consultant episodes (FCEs) in urology, by two urological trainees. The codes were compared with those ascribed by professional hospital coders (and of which the trainees were unaware) from information written on the audit form by junior medical staff. Where there were discrepancies in codes an error was recorded and the stage in the coding process in which it occurred was determined. RESULTS: Forty-eight coding errors were found in 38 of the 106 (36%) FCEs; 34 (71%) were caused by inaccurate coding and 14 (29%) were the result of the incorrect completion of audit forms. CONCLUSION: The clinical codes generated from the authors' department do not accurately reflect the clinical practice. If coding errors of this magnitude are typical of urology units in general, the concept of hospital performance tables (which will be generated using routine clinical data) is untenable unless data recording is given higher priority. PMID- 10691812 TI - Statistical problems with 'optimal' thresholds in studies of new prognostic factors in urology. PMID- 10691813 TI - Ethical and legal aspects of clinical hydration and nutritional support. PMID- 10691814 TI - Reduction of radiation exposure to patients in the follow-up of shockwave lithotripsy. AB - OBJECTIVES: To assess, in patients undergoing extracorporeal shock wave lithotripsy (ESWL), if a policy of using unilateral X-rays of the kidney, ureter and bladder (hemi-KUB) whenever possible and appropriate during diagnosis and follow-up, was successful in reducing the radiation exposure associated with ESWL. PATIENTS AND METHODS: Two groups of patients of statistically comparable size and demography were assessed retrospectively before and after the implementation of the policy. All had undergone ESWL for radio-opaque upper urinary tract stones and all were finally rendered stone-free. The number and type of all radiological procedures from initial diagnosis of the stone to documented stone-free status were recorded and the dose calculated. RESULTS: The appropriate use of hemi-KUB X-rays resulted in a significant mean reduction of radiation exposure after treatment of 2.28 mSv per patient (P<0.05). Furthermore, as expected, the radiation dose was clearly but not closely correlated with stone size (r = 0.419). CONCLUSIONS: The appropriate use of hemi-KUB X-rays during the follow-up after ESWL is a simple and effective way of significantly reducing the radiation exposure of such patients. PMID- 10691815 TI - The indwelling ureteric stent: a 'friendly' procedure with unfriendly high morbidity. AB - OBJECTIVE: To review the morbidity and complications of ureteric stent insertion and to evaluate specifically the effect of an indwelling ureteric stent on the changes in hydronephrosis after stenting. PATIENTS AND METHODS: In a prospective study, 110 renal units with a stent in place were evaluated in 90 patients. Of the 110 stents, 52 were left in place for 3 months, 23 for 6, 11 for 9, 19 for 12 and five (forgotten stents) for 13-30 months. The patients were followed using plain abdominal X-ray at 1 and 30 days after stenting. They were further followed using ultrasonography and plain films every 3 months until the scheduled date for stent removal or the appearance of complications. RESULTS: Thirty-four patients had fever and bacteriuria after stent insertion. Of the 110 stents, 11 (10%) fragmented and nine (8%) migrated. Seventeen patients complained of flank pain on voiding. In 21 renal units (19%) there was no change in the severity of hydronephrosis, whereas in six (5.5%) hydronephrosis developed or worsened after stenting. CONCLUSION: Although ureteric stenting is undoubtedly an important procedure to relieve ureteric obstruction, the indications for stent insertion should be considered carefully in every patient. The close follow-up of stented patients is valuable for the early detection of morbidity or complications and in such cases the stent should be removed or exchanged as soon as possible. PMID- 10691816 TI - Evidence for the autoregulation of vesical circulation by intravesical potassium chloride and distension in the normal human bladder. AB - OBJECTIVES: To determine the influence of distension and intravesical KCl on vesical blood flow in the normal human bladder. Subjects and methods Nine normal volunteers underwent comparative cystometry (NaCl vs 0.2 mol/L KCl; filling rate 50 mL/min). Peak systolic blood flow velocity (PSBFV) and end-diastolic blood flow velocity (EDBFV) were measured in several intramural arteries at a filling volume of 50 mL and at maximum cystometric capacity (Cmax). For these measurements a colour Doppler unit fitted with an endorectal probe was used. The resistance index (RI) was defined as (PSBFV-EDBFV)/PSBFV. RESULTS: In the presence of NaCl, the mean PSBFV increased significantly from 9 cm/s at 50 mL to 17 cm/s at Cmax (512 mL). Compared with NaCl, KCl induced a significantly higher mean PSBFV at 50 mL (15 cm/s). With increasing distension the rise in PSBFV with KCl filling (Cmax 478 mL) was nearly parallel to that obtained on NaCl filling (mean 22 cm/s); the RI did not change. CONCLUSIONS: Bladder distension and intravesical KCl significantly increased the PSBFV. The unchanged RI indicated a concomitant increase in perfusion rates. Hence, the composition of urine (high potassium concentrations and hyperosmolarity) and its storage govern the autoregulation (independent of cardiac output) of vesical circulation, probably by reflexive pathways. These findings provide further evidence for prevesical arteriovenous shunts. PMID- 10691817 TI - Oxybutynin for detrusor instability with adjuvant salivary stimulant pastilles to improve compliance: results of a multicentre, randomized controlled trial. AB - OBJECTIVE: To test the hypothesis that compliance with oxybutynin would be improved if the severity of dry mouth could be reduced, thus leading to improved urinary symptom response and improved outcome, in a randomized, controlled trial of oxybutynin with or without salivary stimulant pastilles in patients with detrusor instability. PATIENTS AND METHODS: Sixty-seven women with detrusor instability were randomized to a variable dose regimen of oxybutynin with (37) or without (30) salivary stimulant pastilles for 8 weeks. Patients were asked to complete a baseline voiding diary. In weeks 1 and 2, patients were encouraged to adjust the dose of oxybutynin themselves to achieve optimum symptomatic control. A second diary was completed in the sixth week and patients were reviewed at 8 weeks. The outcome measures were the compliance rate, follow-up attendance rate, maximum dose of medication, changes in voiding and incontinence episodes, and changes in severity of urgency and of dry mouth symptoms between the first and sixth week. RESULTS: Of the 67 women, 32 (47%) completed the study; the proportion completing was the same in both groups. Four patients had stopped the medication and there was no difference in the distribution of maximum dosage achieved between the groups. Both groups reported a reduced severity of urgency symptoms and increased severity of dry mouth. There were no differences in reported symptom change between the groups during the study. CONCLUSIONS: The combination of oxybutynin and salivary stimulant pastilles does not improve compliance or symptom relief compared with oxybutynin alone; it does not allow a greater dose of oxybutynin to be tolerated. PMID- 10691818 TI - A randomized double-blind placebo-controlled crossover trial of the efficacy of L arginine in the treatment of interstitial cystitis. AB - OBJECTIVES: To determine, in a double-blind placebo-controlled crossover study, whether L-arginine improves the symptoms of interstitial cystitis (IC), a chronic condition in which nitric oxide (NO) may be important, as previous open pilot studies suggested that L-arginine reduced the pain and frequency associated with IC. PATIENTS AND METHODS: Patients fulfilling the standard diagnostic criteria for IC were randomized to receive L-arginine (2.4 g/day) or placebo for one month. After a 2-week 'washout' period they received the other medication. Patients were assessed at each stage using a validated symptom index, a voiding diary, urine analysis and records of adverse events. Patients were asked about overall efficacy at the close of the study. The results were compared using a t test, with significance indicated at P<0.05. RESULTS: Sixteen (16) patients (mean age 51.3 years) were enrolled; the mean duration of IC was 5.4 years, the IC symptom index score 29.1, their nocturnal frequency 3.5 (voided volume 182 mL) and daytime frequency 12.7 (124 mL). Patients on placebo showed no differences in any recorded variable over the baseline values. L-arginine caused a statistically significant reduction in the overall symptom score of 2.2 over baseline, but there was no difference in voided volume, frequency or nocturia. As there was no significant difference for any variable between L-arginine and placebo, this reduction in score should be regarded with caution. Three patients withdrew because of side-effects (severe headaches, night sweats and flushing). CONCLUSION: Oral L-arginine produces a statistically significant improvement in the IC symptom index in patients with IC, but the effect is small. This effect may not be clinically significant as there were no improvements in the other variables assessed and no significant difference between the response to L arginine and placebo. From these results the use of L-arginine cannot be recommended for treating IC. PMID- 10691819 TI - The changing pattern of mortality and morbidity from radical cystectomy. AB - OBJECTIVES: To examine the morbidity and mortality of radical cystectomy as currently practised, and to compare the findings with historical data. PATIENTS AND METHODS: The operative mortality and early and late complications were recorded in 101 consecutive patients (median age 65 years, range 38-81; 33 aged >70 years) undergoing radical cystectomy between April 1992 and October 1997. Fifteen patients had relapsed after previous radical radiotherapy. RESULTS: The median postoperative stay was 14 days (range 8-44). There were two deaths within 60 days of surgery (of patients aged 46 and 59 years) from respiratory failure and sepsis, respectively. The mortality in the elderly was not more than in other age groups. The early morbidity included two cases of lower limb insufficiency, both in the salvage cystectomy group, where the morbidity was significantly higher than in those undergoing primary cystectomy (chi-squared, P<0.01). Three patients underwent early re-exploration. There were four clinically significant episodes of deep vein thrombosis and two pulmonary emboli that were not fatal. CONCLUSION: As currently practised, radical cystectomy is associated with a lower mortality (<2%) and morbidity than described previously. The added morbidity of salvage cystectomy and the acceptable mortality of primary cystectomy suggests that the treatment of choice for muscle-invasive disease is primary cystectomy, with external beam radiotherapy reserved for those patients unfit for major surgical intervention. Age alone should no longer be considered a contraindication to cystectomy. PMID- 10691820 TI - Surveillance for bladder cancer: the management of 4.8 million people. South-west Urologists. AB - OBJECTIVE: To document the workload of bladder cancer surveillance on the British urologist. Methods Thirty-one consultant urologists serving a population of 4.8 million were sent postal questionnaires eliciting their views on the management of superficial bladder cancer. The number, type and outcome of cystoscopies performed over a 6-week period throughout the region was then assessed prospectively. Results One person in 1450 in the South-west region is undergoing follow-up for bladder cancer. Of the responding consultants, 36% would give a single dose of intravesical chemotherapy within 24 h of resection for a G1/2 pTa tumour and 84% would perform the first check cystoscopy at 3-4 months. Over the 6 week period of the study, 696 cystoscopies were performed; there was considerable variation among centres in the choice of cystoscopy type, with 3-80% being rigid cystoscopies. Overall, there was a positive finding in 31% of the assessments. CONCLUSION: This study documents the practice of a significant number of UK urologists in the management of superficial bladder cancer. There are considerable variations among individuals in the type and timing of check cystoscopy. PMID- 10691821 TI - Diagnosis and prediction of recurrence and progression in superficial bladder cancers with DNA image cytometry and urinary cytology. AB - OBJECTIVE: To investigate the roles of urinary cytology and image cytometric analysis of nuclear DNA ploidy pattern in the diagnosis and prediction of recurrence and/or progression of superficial bladder cancers. PATIENTS AND METHODS: Aliquots of catheterized urine from 92 patients with primary (23) or previous (69) superficial bladder cancers were assessed using urine cytology and image-analysis cytometry independently. RESULTS: Of the 23 primary superficial transitional cell carcinomas (TCCs), 11 (48%) were detected by urinary cytology while 12 (52%) were detected by image-analysis cytometry (P>0.05) and 13 (57%) were revealed by combined cytology and cytometry. Of 42 recurrent superficial TCCs, 29 (69%) were detected by urinary cytology, whilst 19 (45%) were diagnosed by cytometry (P<0.05) and 29 (69%) by combined cytology and cytometry. The degree of ploidy in relation to pathological stage and/or grade showed an increasing frequency of aneuploid pattern in more invasive and undifferentiated tumours, but with no statistical significance (P>0.05). The positivity of DNA image cytometry had no significant association (P>0.05) with tumour recurrence and/or progression. CONCLUSIONS: DNA image cytometry can provide a limited but not significant advantage over urinary cytology in the detection of primary superficial TCCs, but it does not seem to be indicated for the prediction of tumour recurrence and/or progression. PMID- 10691822 TI - Does endoscopic laser ablation of the prostate stand the test of time? Five-year results from a multicentre randomized controlled trial of endoscopic laser ablation against transurethral resection of the prostate. AB - OBJECTIVE: To determine the long-term objective and subjective outcome of patients with benign prostatic enlargement (BPE) treated by endoscopic laser ablation of the prostate (ELAP), as part of a multicentre randomized controlled trial of ELAP against TURP. PATIENTS AND METHODS: Initially, 151 patients with BPE were randomized to undergo either ELAP or TURP, starting in March 1992. ELAP was performed using the Urolasetrade mark fibre (Bard, Covington, GA, USA) in conjunction with a Nd:YAG laser source. All patients who had originally participated in the study were approached 5 years later to obtain a urological history, American Urological Association (AUA) symptom score and two measurements of urinary flow rate, with an ultrasonographic assessment of the postvoid residual urine volume (PVR). RESULTS: The mean duration of follow-up was 61 months; 109 patients were traced, comprising 69 who were alive and well, and had undergone no further bladder outlet surgery, 26 who had required revision surgery, 12 who were dead or terminally ill and three who had dementia. Both ELAP and TURP produced sustained improvements in mean AUA score, maximum flow rate and PVR, with respective values at 5 years of 6.3, 17.8 mL/s and 76 mL, and 6.5, 20.0 mL/s and 55 mL. Eighteen of 47 ELAP patients (38%) and eight of 51 (16%) TURP patients underwent revision surgery within the follow-up. CONCLUSION: ELAP and TURP produced similar subjective and objective outcomes at 5 years. The re operation rate after ELAP was more than double that after TURP and suggests that ELAP should not be used routinely in the management of men with BPE. PMID- 10691823 TI - The development and validation of a quality-of-life measure to assess partner morbidity in benign prostatic enlargement. AB - OBJECTIVE: To investigate morbidity in the partners of patients with benign prostatic enlargement (BPE) by developing and validating a disease-specific questionnaire. PATIENTS AND METHODS: Phase 1 of the study comprised preliminary interviews with 15 patients who had newly diagnosed BPE, and with their partners, to determine the relevant issues for the partners. In phase 2, using these issues, a questionnaire was produced and tested on the 15 partners. In phase 3 the questionnaire was completed by 90 further partners, 50 at interview and the next 40 by post. As part of the validation process, the partners were also asked to complete the Short-Form-36 questionnaire, and the patients the Internation Prostate Symptom Score (IPSS) and the ICSmale questionnaires. RESULTS: In phase 1 all 15 of the partners were affected by the patients' disease and nine issues were identified. In phase 2, of the 90 partners, only one had no morbidity from the patient's symptoms; 71% were worried that the patient may have cancer and 69% concerned that the patient may require an operation. Only six partners were present at the patients' urological consultation. The partners' questionnaire scores were related significantly to the Mental Health and Vitality domains of the SF36 and with the patients' IPSS. CONCLUSIONS: The study confirmed the presence of significant morbidity in the partners of patients with BPE. The degree of partner morbidity was related to the severity of the patients' symptoms. Many of the questionnaire issues can be addressed in the consulting room by open discussion with patient and partner. PMID- 10691824 TI - The pharmacovigilance of tamsulosin: event data on 12484 patients. AB - OBJECTIVE: To determine drug effectiveness and adverse effects in a noninterventional observational cohort study of over 10 000 patients treated with tamsulosin in general medical practice. METHODS: Using prescription-event monitoring, data were collected of all prescriptions for tamsulosin issued nationally during June 1996 to January 1998. For each patient entered into the cohort a computerized longitudinal record of exposure was constructed. The outcome data, patient information and an opinion about the effectiveness of the drug were provided by the prescriber, using a standard questionnaire sent 6 months after the initial prescription for tamsulosin. The incidence of each of almost 2000 events listed in the Drug Safety Research Unit computerized dictionary was calculated and scrutinized by medical assessors for possible adverse reactions, and any difference determined between the incidence of each event in the first month and subsequent months of exposure. All deaths were followed up to detect possibly drug-related causes. RESULTS: Event data were obtained on 12484 patients, from the 52.9% of questionnaires returned and that contained valid event data. Tamsulosin was reported to have been effective in 7428 (78.3%) of the 9487 patients in whom the general practitioners expressed an opinion about effectiveness. Suspected adverse drug reactions were reported in only 171 (1.4%) of the cohort. Dizziness, headache, malaise and hypotension were common to the reported adverse reactions, reasons for stopping the drug and events of greatest incidence density. None of the 282 deaths that occurred in this elderly cohort were attributed to the drug. CONCLUSION: This study suggests that tamsulosin has a highly acceptable benefit-to-risk ratio. No untoward features not already mentioned in the prescribing guidance were identified. PMID- 10691825 TI - The role of prostaglandin synthesis in prostate cancer. PMID- 10691826 TI - Rising incidence of prostate cancer in Scotland: increased risk or increased detection? AB - OBJECTIVE: To assess the extent to which the increasing incidence of prostate cancer in Scotland can be explained by increased detection, particularly through transurethral resection of the prostate (TURP) and use of the prostate-specific antigen (PSA) test. Subjects and methods This population-based study was confined to men resident in Scotland and aged > or =50 years. Temporal trends were examined in age-specific and age-standardized incidence, mortality and TURP rates, and PSA testing rates during 1981-1996. Also analysed were the geographical variations in age-standardized incidence and mortality rates during two distinct periods, 1984-1986 (before PSA testing) and 1994-1996 (after PSA testing). Finally, incidence rates and relative survival at 5 years were calculated by age group and 5-year periods of diagnosis during 1968-1992. RESULTS: The incidence of prostate cancer in men aged > or = 50 years increased from an age-standardized rate of 142.0 per 100 000 in 1981 to 240.9 in 1996, with the steepest increase occurring between 1992 and 1993. The mortality rate increased similarly until 1993, but was relatively stable thereafter, falling slightly in 1996. In 1981-1988, incidence rates were closely correlated with TURP rates (r = 0.98, P<0.001). In 1989-1996, incidence was closely correlated with PSA testing rates (r = 0.98, P<0.001). By 1994-1996, incidence rates varied substantially between Scottish mainland health boards (range 167.7-303.0 per 100 000), with much less variation in mortality rates (90.7-110.0). Relative survival has increased recently in all age groups although, in the era before PSA testing, survival was reasonably stable despite increasing incidence. CONCLUSION: Although there may have been a true increase in risk, much of the observed increase in the incidence of prostate cancer in Scotland between 1981 and 1996 has been caused by increased detection, leading recently to considerable variation among different areas of the country. The extent to which this represents the early diagnosis of tumours which would eventually cause symptoms or be life-threatening, or detection of latent disease which would never have become symptomatic, is not clear. There is no evidence so far that the increased incidence is associated with any substantial reduction in mortality. PMID- 10691827 TI - Trends in the curative treatment of localized prostate cancer after the introduction of prostate-specific antigen: data from the Rotterdam Cancer Registry. AB - OBJECTIVE: To investigate changes in the incidence and treatment of prostate cancer over the period in which new diagnostic tools were introduced and the attitude towards treatment was changing. PATIENTS AND METHODS: Information on the extent of disease and treatment of patients diagnosed with prostate cancer within the Rotterdam region was retrieved from the Rotterdam Cancer Registry. RESULTS: In the period 1989-95, 4344 patients were diagnosed with prostate cancer and the age-standardized incidence increased from 62 to 125 per 100 000 men. This increase mainly comprised tumours localized to the prostate, while the incidence of advanced cancers remained stable. The proportion of poorly differentiated tumours decreased from 33% in 1989 to 24% in 1995. In the same period the number of patients receiving radiotherapy increased from 80 to 258, while the annual number of radical prostatectomies rose from 17 to 159. Radiotherapy was the preferred type of treatment in patients over 70 years of age, whereas radical prostatectomy was used more frequently in younger patients with localized tumours. CONCLUSION: While the value of screening for prostate cancer remains in debate, incidence and treatment patterns are changing rapidly. Information on patterns of care is needed to interpret future mortality data and to plan resources for adequate health care. PMID- 10691828 TI - PC-SPES, a dietary supplement for the treatment of hormone-refractory prostate cancer. AB - OBJECTIVE: To assess the effectiveness of PC-SPES, a dietary supplement containing eight herbal extracts, which is a popular alternative therapy among patients with hormone-refractory prostate cancer; anecdotal reports claim that this agent provides relief of metastatic pain, improvements in quality of life and reduction of prostatic specific antigen (PSA) level. PATIENTS AND METHODS: Sixteen men treated for advanced metastatic prostate cancer (stage D3) with either orchidectomy or a luteinizing-hormone releasing hormone agonist, with or without anti-androgen, were enrolled into a prospective clinical trial to evaluate the possible toxic and beneficial effects of PC-SPES. After hormone ablative therapy had failed, and with established disease progression, all patients received supplemental treatment with PC-SPES (2.88 g daily) for 5 months. Hormonal therapy was continued throughout the trial to avoid the known withdrawal effect of anti-androgen on PSA levels. RESULTS: The supplemental intake of PC-SPES was associated with significant (P<0.05-0.01) improvements in quality-of-life measures, reductions in patient's pain ratings (P<0.05-0.01), and a decline in PSA levels (P<0.01), with no major side-effects. CONCLUSIONS: These results support the anecdotal reports of the beneficial effects of PC-SPES as a comparable alternative to current management regimens in hormone-refractory prostate cancer. However, no conclusions can be drawn about the long-term effects of this new herbal therapy. PMID- 10691829 TI - The transverse colonic reservoir: the Unicamp technique. AB - OBJECTIVE: To present the results of a continent and nonrefluxing transverse colonic urinary reservoir technique. PATIENTS AND METHODS: Twenty patients who had received high doses of irradiation underwent construction of transverse colonic reservoir as a primary form of urinary diversion. Fourteen patients had a vesicovaginal fistula after definitive radiation therapy for gynaecological tumours and six had radiation therapy for invasive bladder cancer as a definitive treatment. They were followed for a median (range) of 4.5(1-8) years. Intravenous pyelography before diversion showed mild hydronephrosis in 10 patients. RESULTS: After diversion, hydronephrosis improved in four patients and no upper tract deteriorated. All but one of the pouchograms showed no ureteric reflux. All the patients required clean intermittent self-catheterization every 3-4 h. Persistent asymptomatic bacteriuria was present in 14 patients, although clinical urinary tract infections were not reported. A moderate metabolic acidosis was present in 12 patients, but none required treatment. The urodynamic evaluation revealed a median (range) reservoir capacity of 450 (350-600) mL, with no contractions or contractions of <35 cm H2O. CONCLUSION: These results suggest that the Unicamp technique for constructing a transverse colonic reservoir is a safe and effective diversion, and is recommended as an alternative method for patients treated by pelvic irradiation. PMID- 10691830 TI - Artificial sphincter insertion after radiotherapy: is it worthwhile? AB - OBJECTIVE: To determine the influence of radiotherapy on the outcome of artificial urinary sphincter implantation. PATIENTS AND METHODS: A series of 72 men who had an artificial urinary sphincter inserted were reviewed retrospectively, analysing in detail the information from 15 patients with a past history of pelvic radiotherapy. RESULTS: In those who had undergone radiotherapy, the complication rate was higher, both for re-operation (eight of 15) and infection (three); 11 of the 15 patients were continent after surgery, compared with 51 (89%) of the 57 in the unirradiated group. CONCLUSIONS: An artificial sphincter can be inserted after pelvic radiotherapy reasonably successfully, but at the cost of a high complication and re-operation rate. Patients with a previous history of radiotherapy should be informed of the higher risk of surgical revision associated with insertion of the prosthesis. PMID- 10691831 TI - Priapism in adult Nigerians. AB - OBJECTIVES: To review the presentation and treatment of adult Nigerians with priapism. PATIENTS AND METHODS: Thirty-five men (mean age 35 years, range 20-54) were seen over a 12-year period. All patients underwent an immediate modified or conventional cavernospongiosus shunt and were assessed at 2, 6 and 12 weeks after discharge for erection, orgasm and fibrosis of corpora cavernosa. RESULTS: Ten patients initially presented to traditional healers and 13 presented early to qualified medical practitioners, and were managed conservatively. As a result, 21 of the 35 patients presented to the author's hospital 6-10 days after the onset of erection. Many did not realise that priapism was abnormal, or had no money to attend hospital. After surgical treatment detumescence was obtained in all patients and maintained. At 12 weeks, 14 patients had normal erections, 13 reduced erections and eight no erections. A local aphrodisiac was identified as the commonest predisposing factor, followed by sickle-cell disease. CONCLUSION: This study shows clearly that even if a patient with priapism presents late, vigorous treatment in the form of an adequate shunt should be undertaken as soon as possible, as this is the only chance the patient has to regain potency. PMID- 10691832 TI - The Mitrofanoff procedure: does it last? AB - OBJECTIVE: To examine the long-term effectiveness of the Mitrofanoff principle and establish if the catheterizing channel is sufficiently robust for long-term use. PATIENTS AND METHODS: Ten patients who had undergone Mitrofanoff reconstruction between 1989 and 1991 (minimum follow-up 10 years) were offered reinterview by one of the authors (J.F.), which involved a structured questionnaire assessing catheterization, continence and complications. RESULTS: One patient had died; nine patients were alive and eight agreed to the structured interview. All the patients had their original stoma and all were completely continent. Four of the patients had experienced stenosis, four had had stones and four had been ill with urinary tract infection(s). CONCLUSION: Despite the complications of infection, stones and some episodic stenosis, the Mitrofanoff channel remains functional for long periods without sustaining structural damage. PMID- 10691833 TI - Bladder neck reconstruction in classic bladder exstrophy: the role of osteotomy in the development of continence. AB - OBJECTIVE: To assess the role of osteotomy at the time of bladder neck reconstruction (BNR) for continence in classic bladder exstrophy, in which closure of the pelvic ring and reconstitution of the pelvic diaphragm may affect eventual continence. PATIENTS AND METHODS: The results of using osteotomy at the time of BNR in 29 children were reviewed. The mean bladder capacity before BNR was 76 mL. The indications for osteotomy were a wide pubic diastasis and a soft intersymphyseal bar. After osteotomy, all children were maintained in external fixation and lower-extremity traction for 6-8 weeks. RESULTS: Complications of osteotomy were limited to a partial femoral nerve palsy (one patient) and delayed union of fragments (one patient). Complications of BNR included urethral stricture (five patients) and bladder calculi (six patients). Continence results were modest, with 11 of 29 children (38%) dry during the day (dry interval >3 h) and eight of 29 (28%) dry at night. Eight children had daytime dry intervals of approximately 3 h. The mean preoperative bladder capacity in children who were dry both day and night was 101 mL. CONCLUSIONS: The preoperative bladder capacity remains a key determinant for the attainment of continence after BNR in the reconstruction of classic bladder exstrophy. Osteotomy allows pelvic closure and thus improves cosmesis of the mons and stabilizes the BNR in patients with a soft intersymphyseal bar, but seems to have no effect on continence when performed at the time of bladder neck plasty. PMID- 10691834 TI - Is only meatoplasty a legitimate surgical solution for extreme distal hypospadias? A long-term follow-up after adolescence. AB - OBJECTIVE: To review the long-term outcome of glanular hypospadias repair for extreme distal hypospadias based on simple dorsal Browne meatoplasty and circumcision, with particular attention to patient satisfaction. PATIENTS AND METHODS: A review of medical records identified 124 patients who underwent surgery for hypospadias between 1970 and 1979. Their age at operation ranged from 3 months to 13 years. The operation, performed as a day-case, included dorsal Browne meatoplasty for meatal advancement, based upon the Heineke-Mikulicz principle, using three absorbable sutures and often circumcision. There were no immediate complications. A questionnaire was sent to 111 patients whose present address was available. Two patients had died since the operation and of the 109 remaining patients, 43 (39%) replied. RESULTS: The patients were aged 16-31 years at the time of assessment, giving a follow-up of 18 (13-23) years. Seven of the 43 patients were married. Seven patients had complained of some difficulties with urination, four reporting a poor urinary flow. The stream was directed forward in 32 patients and to some degree downwards in 11, still enabling all the patients to urinate while standing with a single stream. Thirty-two patients had had sexual experience, with no functional problems. Twenty-three defined their penile appearance as normal and 20 as abnormal. Overall, 24 (56%) reported complete satisfaction, 11 (25%) partial satisfaction and eight (19%) were dissatisfied with the surgical result. CONCLUSION: As there are few published reports about the long-term outcome and patient satisfaction after hypospadias repair, it is difficult to draw clear conclusions. However, this simple procedure is a legitimate surgical option for extremely distal glanular hypospadias. PMID- 10691835 TI - Comparison of marker protein expression in benign prostatic hyperplasia in vivo and in vitro. AB - OBJECTIVE: To use multiple immunofluorescence to compare the in vivo and in vitro expression of tissue-specific proteins in BPH. Materials and methods Pure populations of prostate epithelial and stromal cells were produced using standard methods. Serum-free media for epithelial cells were compared. Co-localization of proteins was compared in frozen-tissue sections and cultured cells by simultaneous multiple immunofluorescence, and recorded using a high-resolution charge-coupled device camera. RESULTS: In contrast to the other serum-free media tested, epithelial cells grew without squamous differentiation or vacuolation in prostate epithelial growth medium (PrEGM, Clonetics, BioWhittaker UK Ltd., Berks, UK). These cells were predominantly of a basal phenotype, with some cells showing a luminal phenotype. Most of the stromal cells had features of myofibroblasts, but smooth muscle cells and fibroblasts also were present. CONCLUSION: PrEGM is a commercially available serum-free medium in which primary cultures of prostate epithelial cells can be propagated reproducibly. This study provides a comprehensive description of tissue-specific protein expression in BPH in vivo and in vitro. The use of simultaneous multiple immunofluorescence to study co localization has resulted in a more precise definition of phenotype than has previously been possible, thereby establishing the relevance of the in vitro model system BPH. PMID- 10691836 TI - Analysis of CD44 isoform v10 expression and its prognostic value in renal cell carcinoma. AB - OBJECTIVE: To examine the expression of CD44 isoform v10 (CD44v10) in patients with renal cell carcinoma (RCC), analyse its role in RCC and its relationship with conventional clinical-histopathological experience. Materials and methods Sixty-four RCC specimens and five metastatic specimens were analysed immunohistochemically using a CD44v10 specific antibody. The expression of CD44v10 was compared with the histological grade and clinical or pathological stage of the tumours. RESULTS: Of the 64 primary tumour specimens, 22 (34%) expressed CD44v10 protein; all of these positive specimens were clear-cell and mixed-cell RCC. Staining was also positive in four of five metastatic specimens and negative in all four cases of granular cell carcinoma. CD44v10 expression was significantly correlated with the histological grade (P<0.0001), clinical stage (P = 0.0050) and pathological stage (P = 0.0143) of the tumours. The prognosis for patients with clear-cell RCC who were CD44v10-positive was worse than for patients who were CD44v10-negative (P<0.0001). In subgroups with different tumour stage (< or =pT2 or > or =pT3), the prognosis for patients with positive CD44v10 expression was also worse than for those with no expression (P<0.05). CONCLUSION: The expression of CD44v10 correlated significantly with histological grade, clinical and pathological stage, and with survival in patients with clear-cell RCC. CD44v10 protein may play a role in the progression of clear-cell and mixed cell RCC, and thus the analysis of CD44v10 expression may provide useful prognostic information. PMID- 10691837 TI - Comparative physiology and biochemistry of rat and rabbit urinary bladder. AB - OBJECTIVE: To compare directly the biochemistry and contractile responses of rat and rabbit bladder to different stimuli. Materials and methods Sexually mature male New Zealand White rabbits and Sprague Dawley rats were compared. Each bladder was excised while the animal was anaesthetized; longitudinal bladder strips were cut and then mounted in an organ bath. Tension (2 g) was placed on all strips and each underwent field stimulation (FS) for a total of 20 s at 1-32 Hz, 1 ms and 80 V and was exposed to carbachol (100 micromol/L), ATP (2 mmol/L) and KCl (120 mmol/L). The tension was monitored continually using a polygraph and data stored digitally in a computer. The responses to each stimulus were determined as the maximum tension generated, maximum rate of tension generation and duration to a maximum response. The Ca2+- ATPase activity of the rat and rabbit bladder was determined. Bladder pressures were then predicted from the strip data using Laplace's law and compared with published values. RESULTS: Contractile responses (per unit tissue mass) of rat bladder strips were significantly greater than those of rabbit bladder strips at all frequencies of FS and to carbachol, KCl and ATP. The rate of contractile force generated by rat bladder strips in response to all stimuli were significantly greater than that generated by rabbit strips. Rabbit bladder strips took significantly longer to generate maximum tension than did rat bladder strips in response to pharmacological stimuli. In response to FS, rat strips took significantly longer than rabbit strips to generate maximum tension. Although the predicted rat bladder pressures were significantly greater than those for rabbit, the predicted pressures for both the rat and rabbit were significantly lower than the pressure responses of the isolated whole bladder model. The contractile data correlated well with the Ca2+-ATPase activity data; rat bladder had seven times the enzyme activity of rabbit bladder. CONCLUSION: Per unit mass, rat bladder is capable of generating more than five times the tension of rabbit bladder. Similarly, the rate of tension generation by rat bladder is three to five times greater than that by rabbit bladder. The duration to maximum tension generated in response to FS compared with pharmacological stimuli was affected by the inherent difference in the rate of contractile response to electrical activation compared with agents which diffuse through tissue, and by the difference in size between rat and rabbit bladder smooth muscle cells. PMID- 10691838 TI - Telomerase activity as a potential marker in preneoplastic bladder lesions. AB - OBJECTIVE: To assess telomerase activity (involved in cell immortalization and detectable in most malignant tumours but not in normal somatic tissues) as a marker in cancer diagnosis. PATIENTS AND METHODS: Tissue telomerase activity was assayed by two different techniques, the telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and a telomerase PCR-enzyme linked immunosorbent assay. Malignant and inflammatory bladder lesions and their adjacent normal tissues were assessed for telomerase activity in a group of 18 patients, 14 of whom had urothelial carcinoma and four a nonspecific inflammatory lesion of the bladder. RESULTS: Eleven of the 14 tumour samples analysed were telomerase-positive and two of the three telomerase-negative tumour samples had a detectable 'telomerase inhibitor'. In the apparently normal tissues next to bladder tumours, four of the 14 specimens were telomerase-positive. Interestingly, these lesions were always next to high-grade muscle-invasive bladder tumours (pT2G3). Two of the four nonspecific inflammatory lesions (one of cystitis glandularis and one of severe dysplasia), known to be preneoplastic lesions, were also telomerase-positive. CONCLUSION: These results strongly suggest that the reactivation of telomerase may be an early event in bladder carcinogenesis, preceding morphological changes related to malignant transformation. Telomerase activity may therefore be useful both as an indicator of malignant potential in preneoplastic lesions, e.g. cystitis glandularis and severe dysplasia, and as a prognostic marker of bladder tumour relapse or progression. PMID- 10691839 TI - Gastrinaemia and G-cell density in the antral gastrocystoplasty: an experimental study in rats. AB - OBJECTIVE: To evaluate gastrinaemia and G-cell density in the antrum incorporated into the bladder of rats after antrocystoplasty. Materials and methods Thirty-two adult, female Wistar EPM-1 rats (body weight 200-220 g) were divided into four equal groups that underwent; group 1, no treatment (controls); group 2, a sham operation; group 3, antrectomy; and group 4, antrocystoplasty. The rats were assessed 2 months after treatment, and gastrinaemia and the G-cell density determined in the antrum mucosa incorporated into the bladder. RESULTS: Compared with group 1, serum gastrin was significantly lower in group 3 (P<0.05) and the G cell density lower in group 4 (P<0.05), although there was no decrease in gastrinaemia in group 4. CONCLUSION: Antrocystoplasty in rats did not affect gastrinaemia but reduced the number of G cells in the antrum incorporated into the bladder. PMID- 10691840 TI - Heat-induced apoptosis in human prostatic stromal cells. AB - OBJECTIVE: To determine whether heat, used in transurethral microwave thermotherapy (TUMT) for benign prostatic hyperplasia and which causes necrotic lesions within the adenoma, induces apoptosis in benign human prostatic stromal cells. Materials and methods Prostatic stromal cells were cultured from benign human prostatic tissue. The origin of the cells was identified by immunohistochemical staining and transmission electron microscopy. Cell cultures were exposed to moderate hyperthermia (47 degrees C) for 1 h and any apoptosis detected by light microscopy, transmission electron microscopy and the measurement of induced caspase-3-like activity. RESULTS: The cultures contained a mixed population of smooth muscle cells and myofibroblasts. Twenty-four hours after heat exposure, 76% of the cells were apoptotic and the caspase activity had increased, whereas only 14% of the cells were necrotic. CONCLUSION: Moderate hyperthermia induces apoptosis in cultured human prostatic stromal cells. PMID- 10691841 TI - Prostate selectivity of JTH-601-G1, an active metabolite of JTH-601, in dogs. AB - OBJECTIVE: To evaluate the effect of JTH-601-G1, an active metabolite and glucuronide conjugate of JTH-601 (an alpha1-adrenoceptor antagonist), on smooth muscle contraction in canine prostate and artery, and to examine the effect of JTH-601-G1 on prostatic urethral pressure and blood pressure in anaesthetized dogs. Materials and methods Male beagle dogs were used in both an in vitro and an in vivo study. In the former, the prostate and right common carotid artery were isolated, and smooth muscle strips from the prostate and open-ring strips from the carotid artery prepared. The effects of JTH-601-G1 on phenylephrine- and noradrenaline-induced contraction were assessed in these tissues. In the in vivo study, four dogs were anaesthetized and the change in urethral pressure, blood pressure and heart rate measured continuously. Vehicle (saline) and JTH-601-G1 were then infused intravenously in increasing doses (0.33-3.3 microg/kg/min for 30 min). In three other dogs, the effect of JTH-601-G1 infusion at a higher rate (25 microg/kg/min for 3 h) on blood pressure was evaluated, and the plasma concentration of JTH-601-G1 measured using high-performance liquid chromatography mass spectrometry. RESULTS: Of the distinct metabolites of JTH-601, JTH-601-G1 had the most potent alpha1-adrenoceptor antagonistic effect in isolated canine prostate. JTH-601-G1 also antagonized alpha1-adrenoceptor agonist-induced contraction in common carotid artery, but the pA2 value in the artery was approximately 25 times higher than that in the prostate. In anaesthetized dogs, JTH-601-G1 decreased urethral pressure in a dose-dependent manner; at the highest dose, urethral pressure decreased by 24.5% and blood pressure by 7.0%. However, there was no significant change in heart rate at any dose. The plasma concentration of JTH-601-G1 increased with the dose of JTH-601-G1, but the concentration of both JTH-601 and other metabolites was below the detection limit. The higher JTH-601-G1 infusion rate caused blood pressure to decrease by only 6-10% even at JTH-601-G1 plasma concentrations of approximately 1500 ng/mL during the infusion. Although there was a negative correlation between mean blood pressure and plasma JTH-601-G1 concentration, the decrease in blood pressure was small compared with the reduction in urethral pressure. CONCLUSION: JTH-601-G1 appears to be a major active metabolite of JTH-601 but with a higher selectivity for canine prostate than artery. The results also indicate that in addition to the alpha1A-adrenoceptor, the alpha1L-adrenoceptor plays an important prostatic selective role in smooth muscle contraction via the alpha1-adrenoceptor. PMID- 10691842 TI - Using an instructional model to teach clean intermittent catheterization to children. PMID- 10691843 TI - Development of a urethrorectal fistula after transurethral microwave thermotherapy for benign prostatic hyperplasia. PMID- 10691844 TI - Perivesical inflammation after early mitomycin C instillation. PMID- 10691845 TI - The management of high-risk lymphoblastic leukaemia in children. PMID- 10691846 TI - DDAVP is not a panacea for children with bleeding disorders. PMID- 10691847 TI - Immunoplatelet counting: a proposed new reference procedure. AB - Given the high degree of interoperator error and poor precision of manual platelet counting, it has recently been proposed that an immunoplatelet counting method could become the new reference procedure. Platelets are identified immunologically with a suitable monoclonal antibody, and the platelet count is derived from the ratio of fluorescent platelet events to collected red blood cell (RBC) events that are also counted by a reliable and calibrated standard impedance counter (RBC ratio). In this study, we have set up a rapid and simple method for immunoplatelet counting and simultaneously compared the RBC ratio with the bead ratio derived from two different preparations of commercial calibration beads (Trucount and FlowCount beads). Comparison of the level of imprecision of the RBC ratio with either the manual count or bead ratios revealed a superior coefficient of variation of < 5% even in samples with a platelet count < 20 x 10(9)/l. The RBC ratio correlated extremely well with the existing manual phase reference method (r2 = 0.93) and especially well with three different commercial impedance counters and a dual-angle optical counter (r2 = 0.98-0.99). However, at < 100 x 10(9)/l, the correlation of the RBC ratio with the dual-angle optical count (ADVIA 120) (r2 = 0.96) was superior to all impedance counters. This suggests that automated optical counting methods may be more accurate at determining platelet counts in thrombocytopenic samples. As the RBC ratio is rapid, cheap and relatively easy to perform, we propose that this method could replace the manual count as a new international reference method. PMID- 10691848 TI - Defective fibrinogen polymerization associated with a novel gamma279Ala-->Asp mutation. AB - A woman with menorrhagia was investigated for a suspected fibrinogen mutation when coagulation tests revealed prolonged thrombin (55 s) and reptilase (43 s) times together with a functional and an antigenic fibrinogen concentration of 0.7 and 2.8 mg/ml respectively. Heterozygosity for a gamma-chain mutation was suggested by a doublet gamma band on SDS-PAGE and an increased negative charge was observed on isoelectric focusing of HPLC-isolated gamma-chains. Electrospray ionization mass spectrometry revealed a gamma-chain mass of 48 411 Da, which was 20 Da more than the control value of 48 391 Da. Because the normal and variant gamma-chains were not resolved, this implied a 40-Da increase in 50% of the gamma molecules. An increased negative charge and a 44-Da increase in mass was verified when DNA sequencing showed heterozygosity for an Ala (GCC)-->Asp (GAC) substitution at codon 279 of the gamma-gene. Fibrin polymerization curves indicated a delay in the onset, and a decrease in the rate, of polymerization. Examination of crystal structures showed that the adjacent Tyr-gamma280 side chain is involved in bonding across the D-D interface, and from the proximity of the gamma279Ala-->Asp mutation it would appear that this perturbs the end-to-end DD interactions between fibrin units of the growing polymer. PMID- 10691849 TI - Intracellular accumulation of factor VIII induced by missense mutations Arg593- >Cys and Asn618-->Ser explains cross-reacting material-reduced haemophilia A. AB - Patients with cross-reacting material (CRM)-reduced haemophilia A exhibit reduced levels of factor VIII antigen. In this study, we determined the molecular basis of the genetic defect in the factor VIII gene induced by either the Arg593-->Cys or the Asn618-->Ser missense mutation, identified in two CRM-reduced haemophilia A patients. We introduced either the Arg593-->Cys or the Asn618-->Ser mutation into a B-domain-deleted factor VIII cDNA and expressed the modified cDNAs in C127 cells. Reduced levels of factor VIII activity and factor VIII antigen in conditioned medium of transfected cells indicated that the secretion of both factor VIII variants was impaired. The ratio of factor VIII antigen present in cell extract to that in conditioned medium was 1.9 and 2.4 times higher for rFVIII-R593C and rFVIII-N618S, respectively, than for rFVIII. Metabolic labelling and immunoprecipitation of transfected cells revealed that rFVIII-R593C and rFVIII-N618S persisted somewhat longer inside the cell than factor rFVIII. Intracellular accumulation and subsequent degradation of factor VIII-R593C and factor VIII-N618S may explain the reduced levels of both factor VIII activity and antigen in plasma of mild haemophilia A patients with corresponding genetic defects. PMID- 10691850 TI - A novel polymorphism in intron 1a of the human factor VII gene (G73A): study of a healthy Italian population and of 190 young survivors of myocardial infarction. AB - We have identified a novel polymorphism located in intron 1a of the human factor VII gene, caused by the nucleotide change G to A at position + 73. In a population of 128 healthy individuals from northern Italy, the variant A73 allele had a frequency of 0.21, whereas the frequency of the previously reported 10 bp insertion allele located at -323 in the promoter region was 0.17 and that of the Q353 allele in the catalytic region of the factor VII gene was 0. 20. In 75% of the healthy individuals, the A73 allele was present together with the 10 bp insertion and the Q353 alleles, indicating a strong linkage disequilibrium. The concomitant presence of A73 with both the 10 bp and the Q353 alleles was associated with the lowest factor VII levels, measured as coagulant activity, activated factor VII and factor VII antigen. The G73A polymorphism was also evaluated in 190 survivors of myocardial infarction who had experienced the event before the age of 45 years and in 179 individuals with a negative exercise test matched with patients for sex, age and geographical origin. Patients carrying the A73 allele associated with lower factor VII levels tended to have a lower risk of myocardial infarction (adjusted odds ratio 0.54; 95% confidence intervals 0.29 0.99). In conclusion, we found a novel variant allele in intron 1a of the human factor VII gene that is often associated in healthy individuals with the 10 bp and Q353 alleles in the promoter and catalytic region of the same gene. This intronic mutation, alone or in association with other factor VII gene polymorphisms, might confer protection against myocardial infarction in the young. PMID- 10691851 TI - Combined alpha interferon and ribavirin for the treatment of hepatitis C in patients with hereditary bleeding disorders. AB - Patients with hereditary bleeding disorders who received non-virally inactivated plasma-derived clotting factor concentrates before the mid-1980s invariably became infected with hepatitis C virus (HCV). Therapy with interferon alpha (IFN alpha) alone has been disappointing in this group. We conducted an open-label study, using a combination of IFN-alpha2b (3 million units three times per week) and ribavirin 1-1.2 g/d in 28 patients with hereditary bleeding disorders. Twenty one of the 28 patients had liver biopsy-confirmed chronic hepatitis (median histological activity index 5; range 1-10) and all patients were HCV RNA positive by PCR. Virological response rate to therapy at 3 months was 82% (23 out of 28). Three HIV co-infected patients showed an early virological response with loss of HCV RNA, but two subsequently relapsed after 3 and 6 months of therapy. Four patients stopped treatment early (one at 4, one at 7 and two at 9 months) because of treatment-related side effects, although three of these have maintained a virological response. Seventeen patients completed the 48-week course. Twenty of the 28 (71%) treated have had a durable virological response with a median follow up of 16 months (range 1-24). Combination therapy represents a significant advance in the treatment of hepatitis C in patients with hereditary bleeding disorders. PMID- 10691852 TI - Type 1 von Willebrand disease - a clinical retrospective study of the diagnosis, the influence of the ABO blood group and the role of the bleeding history. AB - This clinical retrospective study investigated the difficulties in diagnosing type 1 von Willebrand disease (VWD). A total of 246 patients previously diagnosed with type 1 VWD were reclassified into 'possible' type 1 VWD (patients with low levels of VWF adjusted for the blood group and either a significant bleeding history or family history) and 'definite' type 1 VWD, requiring low levels of von Willebrand factor (VWF), a bleeding history and inheritance. On reclassification, only 144/246 (59%) patients had low VWF levels adjusted for blood group, 88/246 (36%) patients met all the criteria for 'definite' type 1 VWD and 51/246 (21%) patients were 'possible' type 1 VWD. A significant proportion of patients, 102/246 (42%), remained an indeterminate group with blood type O, VWF levels between 35 and 50 U/dl and personal and/or family bleeding history. This subgroup might require reclassification as 'not VWD'. However, a similar bleeding tendency was found in two matched groups of patients of blood groups O and non-O and VWF levels between 35 and 50 U/dl. These results suggest that the use of ABO adjusted ranges for VWF levels might not be essential for diagnosis, because bleeding symptoms may depend on the VWF level regardless of the ABO type. Of the diagnostic criteria, the bleeding history was of prime importance in the clinical decision to diagnose and treat type 1 VWD. These observations could help in the reconsideration of how the criteria for diagnosing type 1 VWD could be adjusted in order to maximize their clinical relevance. PMID- 10691853 TI - Type II protein C deficiency: identification and molecular modelling of two natural mutants with low anticoagulant and normal amidolytic activity. AB - Two mutations in exons 3 and 9 of the protein C gene were identified by amplification and sequencing from symptomatic probands referred for venous thromboembolism and thrombophilia screening. The phenotype associated with the mutations is a type II protein C deficiency with normal amidolytic activity. In one family, the mutation in exon 3 (G3545-->A), which predicts an R9 to H substitution in the Gla domain, was identified. A mutation in exon 9 (G10899- >A), which predicts an R352 to W substitution in the catalytic site, was identified in the second family and has been reported previously in association with type II deficiency with low amidolytic activity. Western blotting of the purified proteins from the probands' plasma did not show any abnormal migratory pattern. Molecular modelling suggested a possible impairment in the recently described Na+ binding pocket for the R352-->W mutant. No conclusions could be drawn relative to the R9-->H mutant. PMID- 10691854 TI - Prevalence of the post-thrombotic syndrome in young women with previous venous thromboembolism. AB - The prevalence of mild, moderate and severe post-thrombotic syndrome (PTS) among 43 young women with a previous single episode of deep vein thrombosis (DVT) was 67%, 7% and 0% respectively. Subjects were assessed at a mean 51 months after the event. Moderate PTS was more common in women with recurrent (n = 9) DVT (44%, P < 0.001). Chronic venous insufficiency, assessed by light reflection rheography (LRR), was significantly (P < 0.05) more prevalent in women with single previous DVT (n = 40), recurrent DVT (n = 9) and isolated pulmonary embolism (PE) (n = 19) compared with healthy age-matched controls (odds ratios 10.9, 52.4 and 3.8 respectively). LRR findings correlated with moderate, but not mild, PTS. There was no correlation between development of PTS and body mass index. PMID- 10691855 TI - Human osteoblast-like cells and osteosarcoma cell lines synthesize macrophage inhibitory protein 1alpha in response to interleukin 1beta and tumour necrosis factor alpha stimulation in vitro. AB - Recent investigations have demonstrated that macrophage inhibitory protein 1alpha (MIP-1alpha) plays a critical role in haematopoiesis. In part, MIP-1alpha limits the differentiation of early haematopoietic cells, thereby ensuring that sufficient quantities of blood precursors are available to meet haematopoietic demands. MIP-1alpha is produced by cells of the marrow microenvironment (marrow stromal cells) in response to a variety of stimuli, including interleukin 1beta (IL-1beta) and tumour necrosis factor alpha (TNF-alpha). Our recent investigations demonstrated that normal human osteoblast-like cells (HOBs) maintain the early phenotype of haematopoietic precursors, like other members of the bone marrow stroma. Although the precise molecular mechanisms for these observations have not been determined, the production of MIP-1alpha remains one such possibility. In the present study, we investigated whether cells of the osteoblast lineage under basal, IL-1beta and/or TNF-alpha stimulation produce MIP 1alpha. We observed that IL-1beta and TNF-alpha stimulated HOBs and human osteosarcoma cells to rapidly express MIP-1alpha mRNA and to secrete large quantities of the protein. MIP-1alpha mRNA and protein was not, however, detected under basal conditions. Perhaps more importantly, enriched human CD34+ bone marrow cells in co-culture may be capable of stimulating the expression of MIP 1alpha mRNA by HOBs in vitro. These findings suggest that human osteoblast-like cells may produce MIP-1alpha in vivo to support haematopoiesis at sites where osteoblasts and haematopoietic cells are closely associated. PMID- 10691856 TI - Oral magnesium pidolate: effects of long-term administration in patients with sickle cell disease. AB - Prevention of erythrocyte dehydration by specific blockade of the transport pathways promoting loss of potassium (K) is a potential therapeutic strategy for sickle cell (SS) disease. Dietary magnesium (Mg) pidolate supplementation over a 4-week period has been shown to inhibit K-Cl co-transport and reduce dehydration. We report here the results in 17 of 20 patients with SS disease treated in an open-label unblinded study of the effects of long-term (6 months) oral Mg pidolate administration (540 mg Mg/d). A significant decrease (P < 0.0025) was observed with Mg therapy in the distribution widths for red cell mean cell haemoglobin concentration (MCHC) (haemoglobin distribution width; HDW), reticulocyte mean cell volume (red cell distribution width of reticulocytes; RDWr) and MCHC (reticulocyte HDW; HDWr), activity of red cell K-Cl co-transport, Na/Mg exchanger and Ca2+-activated (Gardos) K+ channel, whereas red cell K and Mg contents were significantly increased. Hb levels and absolute reticulocyte counts did not change with Mg therapy. Two patients did not complete the trial because of diarrhoea and one did not complete the trial for unrelated reasons. Although the median number of painful days in a 6-month period decreased from 15 (range 0 60) in the year before the trial to 1 (range 0-18; P < 0.0005) during the period of Mg therapy, no firm conclusion on therapeutic efficacy could be drawn from this unblinded open-label trial. PMID- 10691857 TI - Jamaican Sbeta+-thalassaemia: mutations and haematology. AB - The sickling disorders are a common cause of morbidity and mortality in Jamaica. Sickle cell beta+-thalassaemia is the fourth commonest form, occurring in one in every 3000 births. This is a heterogeneous condition, producing HbS, HbF and HbA2 with variable amounts of HbA, depending on the mutation and, within a defined population, only a few beta-thalassaemia mutations occur at high frequency. This study establishes the frequency of beta-thalassaemia mutations in Sbeta+ thalassaemia patients in Jamaica. In addition, comparison of the haematological phenotypes is possible by looking at the 'average steady-state haematology' of the different mutational groups. Blood samples from 132 unrelated Sbeta+ thalassaemia patients attending the MRC Sickle Cell Unit at the University of the West Indies were analysed by amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) or sequencing to determine the nature and frequencies of the underlying beta-thalassaemia mutations. Ten mutations were identified, four of which accounted for 93% of patients studied. These were -29(A --> G) in 71 (54%), -88(C --> T) in 27 (20%), polyA(T --> C) in 17 (13%) and IVS1 5(G --> C) in nine (7%). The six remaining mutations found at low frequency were C24(T --> A) in two patients and one each of IVS2-848(C --> A), -90(C --> T), IVS1-5(G --> T), IVS1-5(G --> A) and IVS1-6 (T --> C). In one individual, no mutation was found. The three commonest mutations were all associated with haemoglobin levels of greater than 10 g/dl, whereas IVS1-5 (G --> C) had a more severe haematological phenotype. The predominance of -29(A --> G) and -88(C --> T) is in keeping with other studies on populations of African origin. IVS1-5(G - > C) is found chiefly in Indian populations, and all affected families acknowledged Indian ancestry, reflecting the prominent Indian community in Jamaica. PMID- 10691858 TI - Rapid detection of alpha-thalassaemia deletions and alpha-globin gene triplication by multiplex polymerase chain reactions. AB - We describe a sensitive, reliable and reproducible method, based on three multiplex PCR assays, for the rapid detection of seven common alpha-thalassaemia deletions and one alpha-globin gene triplication. The new assay detects the alpha0 deletions - -SEA, - (alpha)20.5, - -MED, - -FIL and - -THAI in the first multiplex PCR, the second multiplex detects the -alpha3.7 deletion and alphaalphaalphaanti3.7 variant, the third multiplex detects the -alpha4.2 deletion. This simple multiplex method should greatly facilitate the genetic screening and molecular diagnosis of these determinants in populations where alpha-thalassaemias are prevalent. PMID- 10691859 TI - Dyserythropoiesis associated with a fas-deficient condition in childhood. AB - Defective lymphocyte apoptosis caused by mutations of the Fas gene can result in an autoimmune lymphoproliferative syndrome (ALPS) in humans. We report two cases of dyserythropoiesis associated with a Fas-deficient condition in childhood. In both cases, dyserythropoiesis predominated on the more mature erythroblasts, and was associated with a lymphoproliferative syndrome as well as with haemolytic anaemia, hypergammaglobulinaemia and the expansion of an unusual population of CD4- CD8- T cells that express the alpha/beta T-cell receptor. The regression of dyserythropoiesis under steroid therapy suggested that it resulted from an autoimmune mechanism, itself secondary to the lymphocyte Fas apoptosis deficiency. Fas-defective apoptosis may be a new aetiology for childhood dyserythropoiesis. PMID- 10691860 TI - Safety profile of the oral iron chelator deferiprone: a multicentre study. AB - In previous trials, the orally active iron chelator deferiprone (L1) has been associated with sporadic agranulocytosis, milder forms of neutropenia and other side-effects. To determine the incidence of these events, we performed a multicentre prospective study of the chelator. Blood counts were performed weekly, and confirmed neutropenia mandated discontinuation of therapy. Among 187 patients with thalassaemia major, the incidence of agranulocytosis (neutrophils < 0.5 x 109/l) was 0.6/100 patient-years, and the incidence of milder forms of neutropenia (neutrophils 0.5-1.5 x 109/l) was 5.4/100 patient-years. All cases of neutropenia resolved after interruption of therapy. Neutropenia occurred predominantly in non-splenectomized patients. Nausea and/or vomiting occurred early in therapy, was usually transient and caused discontinuation of deferiprone in three patients. Mild to moderate joint pain and/or swelling did not require permanent cessation of deferiprone and occurred more commonly in patients with higher ferritin levels. Mean alanine transaminase (ALT) levels rose during therapy. Increased ALT levels were generally transient and occurred more commonly in patients with hepatitis C. Persistent changes in immunological studies were infrequent, although sporadic abnormalities occurred commonly. Mean zinc levels decreased during therapy. Ferritin levels did not change in the overall group but decreased in those patients with baseline levels > 2500 microgram/l. This study characterized the safety profile of deferiprone, and, under the specific conditions of monitoring, demonstrated that agranulocytosis is less common than previously predicted. PMID- 10691861 TI - Excessive apoptosis of bone marrow erythroblasts in a patient with autoimmune haemolytic anaemia with reticulocytopenia. AB - We report a patient with autoimmune haemolytic anaemia (AIHA) with reticulocytopenia, who showed excessive apoptosis of erythroblasts. Ultrastructural analysis of bone marrow cells showed that 50% of erythroblasts had characteristic features of apoptosis, which was confirmed by staining with Annexin-V. In addition, in contrast to normal erythroblasts, Ig staining of the entire erythroblast population could be shown. These data show that apoptosis may contribute to the mechanism of reticulocytopenia in AIHA. PMID- 10691862 TI - High FUS/TLS expression in acute myeloid leukaemia samples. AB - Retinoic acid has the ability to induce differentiation in some myeloid leukaemia cell lines and has been used to induce remission in acute promyelocytic leukaemia patients. We have analysed changes in gene expression, by differential display, in HL60 cells exposed to all-trans retinoic acid (ATRA) for only 1 h. Only about 0.4% of the genes examined by this technique showed changes in expression level, and all four of the gene fragments identified were downregulated during the short 1 h exposure. Two of the fragments were novel, a third was MYC and the fourth was the FUS proto-oncogene. Northern analysis showed that FUS was downregulated within 1 h only during induced neutrophil differentiation but not at all during induced monocyte differentiation. Unlike the sensitive cell lines, ATRA-resistant cell lines did not show a downregulation of FUS over a 24 h period of exposure to ATRA. Using a semiquantitative PCR analysis, no difference in FUS levels was observed between ATRA-sensitive and -resistant cell lines. A similar analysis was carried out on primary acute myeloid leukaemia (AML), peripheral stem cell harvests (PBSC) and cord blood samples. The PBSC and cord blood samples had FUS levels that were similar or generally less than the cell lines. However, much higher levels were seen in 63% of the AML samples examined. The data presented are consistent with previous reports for a role for FUS in the promotion and maintenance of cellular proliferation. PMID- 10691863 TI - Constitutive activation of FLT3 in acute myeloid leukaemia and its consequences for growth of 32D cells. AB - The receptor tyrosine kinase Flt3 is expressed on leukaemic blasts of most cases with acute myeloid leukaemia (AML). In order to evaluate the presence and significance of constitutive activation of Flt3 for leukaemogenesis, we (1) analysed the expression and activation status of the receptor in AML blasts; and (2) evaluated the functional consequences of constitutively active Flt3 in a myeloid progenitor cell line. Immunoprecipitation studies revealed Flt3 expression in a high proportion of AML cases (27/32) with ligand-dependent Flt3 autophosphorylation in 18, constitutive autophosphorylation in three and no autophosphorylation in six cases. Only one out of three samples with constitutively active Flt3 but 3/18 samples with ligand-dependent autophosphorylated Flt3 contained the recently described internal tandem repeat (ITR) mutations. To test the significance of Flt3 activation in myeloid cell function, we also characterized the biochemical and biological effects of the activating mutation D838V of Flt3 (FLt3D838V) on the factor-dependent myeloid progenitor cell line 32Dcl3: cells transfected with wild-type Flt3 (32D/Flt3) grew FLt3 ligand (FL) dependent, and the receptor was ligand dependently autophosphorylated. In contrast, the receptor was constitutively autophosphorylated in 32D/Flt3D838V cells, which grew independently of FL. We conclude that, in some AML samples, Flt3 is constitutively activated and that this does not correlate with ITR mutations in the juxtamembrane domain. Furthermore, constitutively active Flt3 confers factor independence to the myeloid progenitor cell line 32D. It remains to be determined whether activation of Flt3 is leukaemogenic in vivo and whether strategies aimed at inhibition of Flt3 activation could inhibit leukaemogenesis. PMID- 10691864 TI - Cell density-dependent VP-16 sensitivity of leukaemic cells is accompanied by the translocation of topoisomerase IIalpha from the nucleus to the cytoplasm. AB - The resistance of several leukaemic and myeloma cell lines (CCRF, L1210, HL-60, KG-1a and RPMI 8226) to VP-16 was found to increase with cell density and to be maximal (3.5- to 39-fold) in plateau phase cell cultures, as measured by clonogenic and MTT assays. Non-transformed confluent Flow 2000 human fibroblasts and Chinese hamster ovary (CHO) cells were also five- and 15-fold resistant to VP 16 respectively. The transition from log to plateau phase was accompanied by a drastic decrease in topoisomerase (topo) IIalpha content in CHO cells and human fibroblasts, while the leukaemic cells maintained constant cellular levels of topo IIalpha and topo IIbeta. However, the nuclear topo IIalpha content was found to decrease as a result of translocation of the enzyme to the cytoplasmic compartment in the leukaemic cells. This was confirmed by subcellular fractionation experiments, Western blotting analyses and immunocytochemistry studies. The quantity of topo IIalpha in plateau phase cytoplasmic fractions ranged from 18% in L1210 cells to 50% in HL-60 and 8226 cells, as measured by both immunoblotting and quantification of the label in immunofluorescent images. The cytoplasmic fraction from plateau phase cells retained topo II catalytic activity, as measured by the decatenation of kinetoplast DNA. The nuclear cytoplasmic ratio of topo IIalpha may be critical in determining the sensitivity of leukaemic cells to topo II inhibitors. Cytoplasmic trafficking of topo IIalpha was observed in plasma cells obtained from patients with multiple myeloma, and perhaps contributes to drug resistance in this disease. PMID- 10691865 TI - Incidence, characterization and prognostic significance of chromosomal abnormalities in 640 patients with primary myelodysplastic syndromes. Grupo Cooperativo Espanol de Citogenetica Hematologica. AB - Recently, a consensus International Prognostic Scoring System (IPSS) for predicting outcome and planning therapy in the myelodysplastic syndromes (MDS) has been developed. However, the intermediate-risk cytogenetic subgroup defined by the IPSS includes a miscellaneous number of different single abnormalities for which real prognosis at present is uncertain. The main aims of this study were to evaluate in an independent series the prognostic value of the IPSS and to identify chromosomal abnormalities with a previously unrecognized good or poor prognosis in 640 patients. In univariate analyses, cases with single 1q abnormalities experienced poor survival, whereas those with trisomy 8 had a higher risk of acute leukaemic transformation than the remaining patients (P = 0.004 and P = 0.009 respectively). Patients with single del(12p) had a similar survival to patients with a normal karyotype and showed some trend for a better survival than other cases belonging to the IPSS intermediate-risk cytogenetic subgroup (P = 0.045). Multivariate analyses demonstrated that IPSS cytogenetic prognostic subgroup, proportion of bone marrow blasts and haemoglobin level were the main prognostic factors for survival, and the first two characteristics and platelet count were the best predictors of acute leukaemic transformation risk. A large international co-operative study should be carried out to clarify these findings. PMID- 10691866 TI - Cladribine with or without prednisone in the treatment of previously treated and untreated B-cell chronic lymphocytic leukaemia - updated results of the multicentre study of 378 patients. AB - Between January 1992 and January 1999, we treated 378 B-chronic lymphocytic leukaemia (CLL) patients with cladribine (2-CdA), and 255 of the patients were also treated with prednisone. A total of 194 patients were previously untreated, and 184 had relapsed or refractory disease after previous other therapy. Complete response (CR) was obtained in 111 (29.4%) and partial response (PR) in 138 (36.5%) patients, giving an overall response (OR) rate of 65.9%. CR and OR were achieved more frequently in patients in whom 2-CdA was a first-line treatment (45.4% and 82.5% respectively) than in the pretreated group (12.5% and 48.4% respectively) (P < 0.0001). The median duration of OR for previously untreated patients was 14.7 months and for pretreated patients 13.5 months (P = 0.09). The median survival evaluated from the beginning of 2-CdA treatment was shorter in the pretreated group (16.3 months) than in the untreated group (19.4 months) (P < 0.0001). A total of 117 (63.9%) patients died in the pretreated group and 63 (32.6%) in the untreated group. In pretreated patients, 2-CdA + prednisone (P) and 2-CdA alone resulted in similar OR (51.0% and 45.0% respectively; P = 0.4). In contrast, in untreated patients, 2-CdA + P produced a higher OR (85.4%) than 2 CdA alone (72.1%) (P = 0.04). Infections and fever of unknown origin, observed in 91 (49.4%) pretreated and 74 (38.1%) untreated patients (P = 0.03), were the most frequent toxic effects. Our results indicate that 2-CdA is an effective, relatively well-tolerated drug, especially in previously untreated CLL. PMID- 10691867 TI - The diagnosis of low-grade peripheral B-cell neoplasms in bone marrow trephines. AB - The aim of this study was to establish how effective is the use of immunohistochemistry on formalin-fixed bone marrow in diagnosing low-grade B-cell neoplasms. We investigated a series of 41 consecutive patients with bone marrow involvement for whom no other diagnostic tissues were available. The sections were stained with the following antibodies: CD3, CD20, CD79a, CD5, CD10, CD23, anti-cyclin D1 and kappa and lambda light chains. Antigen retrieval was performed using either a microwave oven or a pressure cooker. Labelling was performed with an avidin-biotin-peroxidase labelling system. A final diagnosis was reached in 37 out of 41 cases (90%): B-chronic lymphocytic leukaemia (15 cases), follicular lymphoma (10 cases), mantle-cell lymphoma (eight cases) and lymphoplasmacytoid lymphoma/immunocytoma (four cases). In the remaining four cases, a generic diagnosis of low-grade B-cell neoplasm was made. The immunophenotyping of formalin-fixed marrow is a useful technique for diagnosing most of the low-grade B-cell neoplasms. PMID- 10691868 TI - Recurrent B-cell non-Hodgkin's lymphoma in two brothers with X-linked lymphoproliferative disease without evidence for Epstein-Barr virus infection. AB - We present two male siblings suffering from recurrent manifestations of B-cell non-Hodgkin's lymphoma (NHL) and recurrent infections of the lower respiratory tract associated with bronchiectasis. Immunodeficiency could not be demonstrated by any laboratory investigation. In both patients, lymphomas developed without evidence for Epstein-Barr virus (EBV) infection, i.e. no antibody response to EBV specific antigens, negative EBV-PCR (polymerase chain reaction) in peripheral blood cells, and absence of latent membrane protein (LMP) and EBV-encoded RNA (EBER) in lymphoma cells. Molecular analysis of the SH2D1A, the gene for X-linked lymphoproliferative disease (XLP) led to the identification of a deletion in the first exon in both patients. Therefore, we postulate that the genetic defect and the following dysregulation of the B-/T-cell interaction rendered these patients susceptible to the early onset of B-cell NHL and that EBV infection is not an obligate prerequisite. PMID- 10691869 TI - Human myeloma cells promote the production of interleukin 6 by primary human osteoblasts. AB - Interleukin-6 (IL-6) is an important growth and survival factor for myeloma cells. However, the identity of the cells producing IL-6 in vivo remains unclear. Myeloma cells are found closely associated with sites of active bone turnover, and cells of the osteogenic lineage, including bone marrow osteoprogenitors, osteoblasts and bone lining cells, may therefore be ideally placed to synthesize IL-6. We have examined the possibility that human osteogenic cells may produce IL 6 in response to stimulation by myeloma cells. Primary human osteoblasts (hOBs) were isolated from normal donors, co-cultured with the human myeloma cell lines, JJN-3, RPMI-8226 and NCI-H929, and the amount of IL-6 released was determined by enzyme-linked immunosorbent assay (ELISA). All myeloma cells stimulated a significant increase in the production of IL-6 when cultured with hOBs (P < 0.05). Prior fixation of hOBs completely abrogated release of IL-6 in the co cultures. In contrast, fixed myeloma cells retained the ability to induce IL-6 production, suggesting that hOBs were the principal source of IL-6. Physical separation of myeloma cells from hOBs using transwell inserts caused a partial inhibition of IL-6 release (P < 0.05), whereas the addition of media conditioned by myeloma cells to cultures of hOBs stimulated a significant increase in IL-6 production (P < 0.05). hOBs secreted greater amounts of IL-6 than human bone marrow stromal cells (hBMSCs) (2.2- to 3.5-fold, P < 0.05), but incubating hBMSCs with dexamethasone to stimulate osteoblastic differentiation resulted in an increase in their ability to produce IL-6 (1.7- to 4. 8-fold, P < 0.05) and to respond to myeloma cells (P < 0.05). These data clearly indicate that cells of the osteoblast lineage release significant amounts of IL-6 in response to stimulation by myeloma cells and may contribute to the IL-6 that promotes the proliferation and survival of myeloma cells in vivo. PMID- 10691870 TI - Therapy with thalidomide in refractory multiple myeloma patients - the revival of an old drug. AB - We have treated 17 refractory or relapsed multiple myeloma patients resistant to chemotherapy with thalidomide at a dose of 200-800 mg/day. Eleven patients responded, five of whom had a very good partial response (> 75% decline in M protein) and another five exhibited a partial response (> 50% decline in M protein). Except for one patient, treatment was well tolerated with only mild side-effects. Thalidomide should be included in the therapeutic options for refractory myeloma. PMID- 10691871 TI - Lamivudine allows completion of chemotherapy in lymphoma patients with hepatitis B reactivation. AB - Reactivation of hepatitis B virus in patients receiving chemotherapy for non Hodgkin's lymphoma (NHL) may give rise to hepatitis, hepatic failure and death, and prevent further chemotherapy. We report four patients with NHL in whom hepatitis flare-up was observed after two (three patients) and six (one patient) cycles of chemotherapy. After spontaneous recovery, they were treated with Lamivudine (100 mg/day), which enabled completion of chemotherapy without further hepatitis B reactivation. In one patient, high-dose chemotherapy and autologous stem cell transplantation was also performed. These data suggest a possible role for Lamivudine in preventing hepatitis B reactivation during chemotherapy administration to chronic carriers of the hepatitis B virus. Moreover, it enabled the completion of both standard and high-dose chemotherapy in patients with previous hepatitis B reactivation. PMID- 10691872 TI - T-prolymphocytic leukaemia with spontaneous remission. AB - T-prolymphocytic leukaemia (T-PLL) is a rare dis-order with a poor prognosis. A 69-year-old man was diagnosed as having a small-cell variant of T-PLL according to the French-American-British classification by haematological, immunological and ultrastructural studies, although the cells had a CD7- phenotype and no chromosomal abnormality. He had no symptoms or organomegaly. The number of his lymphocytes, 53.7 x 109/l at the time of diagnosis, gradually decreased without therapy, and he was in complete remission 39 months later. A rearranged band in the T-cell antigen receptor-beta gene, which was detected at the time of diagnosis, decreased or disappeared. This is the first report of a T-PLL case with spontaneous complete remission. PMID- 10691873 TI - Second allogeneic haematopoietic stem cell transplantation in relapsed acute and chronic leukaemias for patients who underwent a first allogeneic bone marrow transplantation: a survey of the Societe Francaise de Greffe de moelle (SFGM). AB - Although recurrent malignancy is the most frequent indication for second stem cell transplantation (2nd SCT), there are few reports that include sufficiently large numbers of patients to enable prognostic factor analysis. This retrospective study includes 150 patients who underwent a 2nd SCT for relapsed acute myeloblastic leukaemia (n = 61), acute lymphoblastic leukaemia (n = 47) or chronic myeloid leukaemia (n = 42) after a first allogeneic transplant (including 26 T-cell-depleted). The median interval between the first transplant and relapse, and between relapse and second transplant was 17 months and 5 months respectively. After the 2nd SCT, engraftment occurred in 93% of cases, 32% of patients developed acute graft-vs.-host disease (GVHD) >/= grade II and 38% chronic GVHD. The 5-year overall and disease-free survival were 32 +/- 8% and 30 +/- 8%, respectively, with a risk of relapse of 44 +/- 12% and a transplant related mortality of 45 +/- 9%. In a multivariate analysis, five factors were associated with a better outcome after 2nd SCT: age < 16 years at second transplant; relapse occurring more than 12 months after the first transplant; transplantation from a female donor; absence of acute GVHD; and the occurrence of chronic GVHD. The best candidates for a second transplant are likely to be patients with acute leukaemia in remission before transplant, in whom the HLA identical donor was female and who relapsed more than 1 year after the first transplant. PMID- 10691874 TI - CD34+ cell dose predicts relapse and survival after T-cell-depleted HLA-identical haematopoietic stem cell transplantation (HSCT) for haematological malignancies. AB - Seventy-eight patients with haematological malignancies, received T-cell-depleted stem cell transplants and cyclosporin followed by delayed add-back of donor lymphocytes to prevent leukaemia relapse. The source of stem cells was bone marrow in 50 patients and granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood in 28 patients. In univariate analysis, only the CD34+ cell dose (but not the stem cell source or the T lymphocyte dose) and disease status were predictive for transplant-related mortality, relapse and survival. Patients receiving >/= 3 x 106 CD34+ cells/kg had an overall actuarial survival of 68% compared with 52%, 35% and 10%, respectively, for cell doses of 2-2.99, 1-1.99 and < 1 x 106/kg. Multivariate analysis of risk factors for relapse identified disease risk and CD34+ cell dose as the only factors. Relapse was 62.5% in 38 patients at high risk of relapse vs. 25% for 40 patients at intermediate or low risk. CD34+ cell doses of >/= 3 x 106/kg were associated with a 13.5% relapse vs. 48% for recipients of lower doses. This favourable effect of CD34+ cell dose on relapse was apparent in both high- and intermediate- plus low-risk groups. Our results support the potential benefit of a high stem cell dose in lowering transplant-related mortality (TRM) and in reducing relapse after allogeneic marrow or blood stem cell transplants. PMID- 10691875 TI - Reconstitution of the cellular immune response after autologous peripheral blood stem cell transplantation in patients with non-Hodgkin's lymphoma. AB - Peripheral blood stem cell (PBSC) transplants may be depleted of lymphoid progenitors, thereby disabling the cellular immune response against viral pathogens after autologous PBSC transplantation (PBSCT). To monitor the cellular immune reconstitution after autologous PBSCT, we investigated the cytolytic activity (CLA) of peripheral blood T lymphocytes against Epstein-Barr virus (EBV) in 13 patients with non-Hodgkin's lymphoma or multiple myeloma. The individual EBV-directed CLA (EBV-CLA) was determined by calculating the number of cytolytic effector cells in 106 T cells needed to lyse 25% of autologous EBV-transformed B lymphoblastoid cells, expressed as lytic units (LU25). During the first 6 months after PBSCT, the EBV-CLA was only 14.6% of the response of healthy controls (median 4. 8 vs. 32.9 LU25). Thereafter, the EBV-CLA increased to 28.15 LU25 (median) or 86% of healthy controls. Monthly follow-up analyses in five selected patients showed that the EBV-CLA was barely detectable at 4 weeks and recovered at 8-12 weeks after PBSCT in four out of five patients. Effector cells consisted mostly of CD8-positive T lymphocytes, with small CD4- and CD3/CD56-positive lymphocyte fractions. These results suggest that the reconstitution of the cellular immune response against EBV takes 8-12 weeks after autologous PBSCT. PMID- 10691876 TI - Incubation of murine bone marrow cells in hypoxia ensures the maintenance of marrow-repopulating ability together with the expansion of committed progenitors. AB - We developed previously a hypoxic culture system in which progenitors endowed with marrow-repopulating ability (MRA), unlike committed progenitors, were selected and maintained better than in air. We report here an improvement to this system targeted at combining the maintenance of progenitors sustaining MRA with the numerical expansion of multipotent and committed progenitors. Murine bone marrow cells were incubated at 1% oxygen in liquid medium supplemented with stem cell factor, granulocyte colony-stimulating factor, interleukin-6 and interleukin 3. In day 8 hypoxic cultures, the numbers of high proliferative potential and granulocyte/macrophage colony-forming cells (HPP-CFC and CFU-GM) were increased with respect to time zero. Colonies generated by HPP-CFC derived from hypoxic cultures exhibited a high replating ability, whereas colonies generated by HPP CFC derived from control cultures exhibited a low replating ability. MRA was fully maintained in hypoxia and markedly reduced in air. Thus, severe hypoxia is able to ensure a full maintenance of progenitors sustaining MRA, together with a significant expansion of in vitro-detectable clonogenic progenitors, including those endowed with replating ability. This system could contribute to the improvement of current techniques for the in vitro treatment of human haematopoietic cell populations before transplantation. PMID- 10691877 TI - Induction of remission after donor leucocyte infusion for the treatment of relapsed chronic idiopathic myelofibrosis following allogeneic transplantation: evidence for a 'graft vs. myelofibrosis' effect. AB - A 54-year-old man showed evidence of disease progression and a reduction in donor chimaerism by molecular microsatellite analysis 6 months after an allogeneic peripheral blood stem cell transplant for chronic idiopathic myelofibrosis. He was treated with a single infusion of donor leucocyte infusions (DLI), which led to the development of mild acute graft versus host disease (GVHD) and the rapid restoration of full donor haemopoiesis. This subsequently led to a progressive reduction in marrow fibrosis from grade IV to grade I over the following 6 months. We believe that this is the first report to provide clear evidence for the efficacy of DLI in this setting, which also provides evidence for the existence of a T-cell-mediated 'graft vs. myelofibrosis' effect similar to that seen against other haematological malignancies. PMID- 10691878 TI - A study of the iron and HFE status of blood donors, including a group who failed the initial screen for anaemia. AB - A complete data set (age, weight, diet and recent donation history; venous blood cell count, serum ferritin and soluble transferrin receptor concentrations and transferrin saturation; HFE genotype) was obtained from 113 male and 122 female blood donors. Progressive iron depletion and deficiency - most apparent from serum concentrations of soluble transferrin receptor divided by the logarithm of ferritin concentrations (the TfR-F index) - developed in men donating up to six times in 2 years, although the serum ferritin alone was also informative; however, no prediction could be made for those iron-depleted individuals who will develop iron deficiency after donation. Iron stores in the groups of donors with 'low-normal' haemoglobin (Hb) concentrations were indistinguishable from those in donors with higher Hb values, whereas donors failing the anaemia screen had reduced stores. This supports the UK policy of accepting donations from people whose Hb concentration is up to 0. 5 g/dl below the recommended European threshold. Women eating red meat once a week sustained higher ferritin concentrations, and the iron status of first-time women donors resembled that of men donating twice each year. Homozygosity for either HFE variant allowed greater iron retention in the face of regular donation, but among heterozygotes the findings were inconclusive. PMID- 10691879 TI - A rapid one-stage whole-blood HPA-1a phenotyping assay using a recombinant monoclonal IgG1 anti-HPA-1a. AB - Severe neonatal alloimmune thrombocytopenia and patients with HPA-1a-specific antibodies require transfusion of HPA-1a-negative platelets. Identifying HPA-1a negative donors requires simple and reliable typing methods. Most existing techniques use polyclonal antibodies, are time consuming and involve platelet isolation. We have used a horseradish peroxidase (HRP)-conjugated recombinant IgG1 anti-HPA-1a (CAMTRAN007) to develop a rapid and reliable enzyme-linked immunosorbent assay (ELISA), which eliminates sample preparation and reduces the incubation and wash steps associated with traditional sandwich ELISAs. The assay uses simultaneous incubation of the monoclonal antibody RFGP56 to capture GPIIbIIIa from whole blood and the recombinant IgG1 antibody to detect captured HPA-1a antigen. It allows 96 samples to be typed in less than 1 h and can be used on stored samples. Initial testing of 85 samples of known HPA-1a genotype demonstrated that HPA-1a-negative samples had OD values of < 0.266, whereas HPA 1a-positive samples had OD values of > 0.6. Testing of 1862 random donor samples in two blood centres confirmed these OD cut-off values and identified 45 HPA-1a negative samples (2.4%), all except one giving OD values of < 0.2. The remaining HPA-1a-negative sample had an OD value of 0.303. The HPA-1a status on all the negative samples and an equivalent number of randomly selected positive samples was confirmed by flow cytometry and polymerase chain reaction with sequence specific primers (PCR- SSP). PMID- 10691880 TI - New genotypes in Fy(a-b-) individuals: nonsense mutations (Trp to stop) in the coding sequence of either FY A or FY B. AB - Duffy blood group antigens are carried on a glycoprotein that is predicted to pass through the erythrocyte membrane seven times and is a promiscuous chemokine receptor. The Fy(a- b-) phenotype is present in two-thirds of African-American Blacks but is rare in Caucasians. In Blacks, the phenotype is due to a non functional GATA-1 motif in the FY B, which silences the gene in erythrocytes but not in other tissues, and these patients do not generally make anti-Fyb or anti Fy3. We describe here the molecular analysis of FY in three unrelated Caucasians who were studied because they had strong anti-Fy3 in their serum. Each was found to have a point mutation that was predicted to change a tryptophan to a premature stop codon in the coding sequence. In one patient (patient 1), the nonsense mutation was at nucleotide 287 of the major transcript in FY A; in another (patient 2), it was at nucleotide 407 in the major transcript of FY B; and in a third (patient 3), it was at nucleotide 408 of the major transcript of FY A. PMID- 10691881 TI - Prevention of worsening of severe thrombocytopenia after red cell transfusions by the use of leucocyte-depleted blood. AB - Platelet counts were measured before and after red cell transfusions in 30 patients with anaemia and severe thrombocytopenia resulting from haematological diseases. There was a mean reduction of 1.1 x 109/l (P = 0.43) in the platelet count after transfusions of 2-3 units of leucocyte-depleted red cell concentrates (20 patients). However, there was a mean reduction of 2.7 x 109/l (P = 0.03), approximately 10%, in the platelet count after transfusions of non-leucocyte depleted red cell concentrates (10 patients). The findings suggest that the forthcoming introduction of universal leucocyte depletion of red cell concentrates will minimize the worsening of thrombocytopenia that occurs in severely thrombocytopenic patients receiving standard non-leucocyte-depleted red cell concentrates. PMID- 10691882 TI - Comparing near misses with actual mistransfusion events: a more accurate reflection of transfusion errors. AB - In a retrospective review of transfusion errors in a large teaching hospital, we found the true incidence of errors to be at least four times the actual mistransfusion events detected. Seventy-five per cent of the errors were detected as near misses. The mistransfusions equated to 1/8610 compatibility procedures, and 1/27 007 units of blood issued, whereas the number of true transfusion errors equates to 1/2153 compatibility procedures and 1/6752 units of blood issued. The major error-prone activities included patient identification at phlebotomy and the final infusion of the blood product at the bedside. Of the cases, 95.2% were due to non-compliance with existing guidelines. Potential disasters were avoided only by the vigilance of the blood bank staff and the systems in place to detect errors. PMID- 10691883 TI - Changes in interferon-gamma production following specific allergen immunotherapy: biology vs methodology. PMID- 10691884 TI - Fibroblasts: a cell type central to eosinophil recruitment? PMID- 10691885 TI - Immunoglobulin E and allergic disease in Africa. PMID- 10691886 TI - Oilseed rape - allergen or irritant? PMID- 10691887 TI - Airway inflammatory and antioxidant responses to oxidative and particulate air pollutants - experimental exposure studies in humans. PMID- 10691888 TI - gammadelta T cells in allergic airway diseases. PMID- 10691889 TI - Complementary DNA cloning and expression of a newly recognized high molecular mass allergen phl p 13 from timothy grass pollen (Phleum pratense). AB - BACKGROUND: Grass pollen extracts contain a range of different allergenic components that can be classified as having low, middle or high molecular mass. Almost 75% of patients allergic to grass pollen display immunoglobulin (Ig) E reactivity to allergens in the high molecular mass range of 55-60 kDa. These proteins have not yet been fully characterized on the protein and DNA level. OBJECTIVE: The aim of this study was to identify and characterize an allergen of the high molecular mass fraction of Phleum pratense pollen by N-terminal protein sequencing and molecular cloning. METHODS: A previously uncharacterized allergen which migrates as a double band with a molecular mass of 55-60 kDa was biochemically purified and investigated by N-terminal sequencing. Subsequently, a DNA primer was designed to amplify the corresponding cDNA using PCR. The cloned cDNA and deduced amino acid sequence were compared with sequence data bases. Immunoblots carrying the recombinant expression product were developed with monoclonal antibodies and sera derived from allergic subjects. The IgE-binding capacity of natural and recombinant allergen was determined using EAST. RESULTS: The nucleic acid sequence as well as the deduced amino acid sequence consisting of 394 amino acids indicated homology with pollen specific polygalacturonases. Four potential sites for glycosylation and 16 cysteine residues were found. The recombinant expression product exhibited the same molecular size as the natural allergen and was clearly IgE-reactive. CONCLUSION: The newly characterized allergen Phl p 13, which shows homology with polygalacturonases, is clearly different from the allergen designated as Phl p 4 and therefore the high molecular mass fraction is composed of at least two different allergens. A possible reason why this important allergen has not been detected until now is that Phl p 13 and Phl p 4 are hardly separable by one dimensional SDS-PAGE. PMID- 10691890 TI - Allergen-specific production of interferon-gamma by peripheral blood mononuclear cells and CD8 T cells in allergic disease and following immunotherapy. AB - BACKGROUND: CD8 T cells are important immunoregulatory cells in animal models of allergic disease, but their role in human allergic immune responses has not been defined. With the development of novel immunotherapeutic reagents, it is clearly important to ascertain whether CD8 T-cell responses are altered following conventional allergen-specific immunotherapy (SIT) and hence targets for future developments/strategies. OBJECTIVE: To study the allergen-specific cytokine release of freshly isolated CD8 T cells from the blood of separate groups of house dust mite- (HDM) allergic patients, patients post-SIT and control nonatopic donors. METHODS: CD8 T cells were isolated by positive selection with immunomagnetic beads and cultured with the affinity purified major mite allergen Der p 1 or with different mitogens, using irradiated autologous peripheral blood mononuclear cells as antigen-presenting cells (APCs). Supernatants were collected at a number of time points and assayed by ELISA for the cytokines interleukin (IL) -4, IL-5 and interferon-gamma (IFNgamma). RESULTS: CD8 T cells stimulated with Der p 1 produced significant quantities of IFNgamma with cells from HDM allergic subjects releasing considerably more IFNgamma than cells from nonatopic subjects, an average of 804 +/- 283 pg/mL of supernatant compared with 30.2 +/- 18.8 pg/mL (P = 0.006). The cytokine was detected in cultures of 16/17 of the allergic subjects and 4/7 of the nonatopic. CD8 T cells from 6/10 patients who had received HDM-SIT released IFNgamma at an average of 363 +/- 202 pg/mL, which was less than the allergic group but still higher than the nonatopic (P = 0.05). Equivalent levels of IFNgamma were detected when the cells were stimulated with the mitogen PHA and this was the same in all groups. Reliable allergen-specific release of significant quantities of IL-4 or IL-5 was not detected from CD8 T cells. CONCLUSION: Allergen-specific IFNgamma is produced at far greater levels from CD8 T cells of HDM-sensitive allergic patients than from nonatopic control individuals and this level is reduced following SIT. PMID- 10691891 TI - T-cell cytokine pattern at three time points during specific immunotherapy for mite-sensitive asthma. AB - BACKGROUND: Several lines of evidence indicate that specific immunotherapy may act by modifying the patterns of cytokines produced by helper T cells. However, different protocols have been used and different results obtained. OBJECTIVES: To quantify the effect of specific immunotherapy on the TH1/TH2 T-cell cytokine pattern at the single cell level. METHODS: We examined the interferon gamma/interleukin-4 ratio in peripheral blood CD4+ and CD8+ T cells from 12 subjects with house dust mite-sensitive asthma using a flow cytometric method of intracellular cytokine detection. Cytokine production was determined following stimulation with phorbol myristate acetate/ionomycin, a policlonal activator. Subjects were examined at three occasions: before specific immunotherapy, after the 3-months dose increase phase and after 1 year of treatment. During the treatment year patients kept a diary in which they recorded: (a) symptoms of asthma according to a 0-3 grading (0 = absent, 1 = mild, 2 = moderate, 3 = severe); (b) number of puffs (100 microg) per day of salbutamol required to control symptoms; and (c) peak expiratory flow. RESULTS: Specific immunotherapy improved clinical indices of disease activity including symptom scores and medication use during the treatment year, and had a marked effect in increasing the interferon-gamma/interleukin-4 ratio in peripheral blood CD4+ T cells already after the dose increase phase (5.47 +/- 1.5 vs 4.07 +/- 1.49%, P = 0.03) with and a further rise after 1 year's treatment (16.12 +/- 2.8 vs 4.07 +/- 1.49 and 16.12 +/- 2.8 vs 5.47 +/- 1.5%, P = 0.001 and P = 0.002, respectively). There were no significant changes in the interferon-gamma/interleukin-4 ratio in peripheral blood CD8+ T cells at the three times of the study. CONCLUSIONS: These data add to view that the efficacy of specific immunotherapy may be attributed to a modified cytokine secretion of CD4+ T cells. PMID- 10691892 TI - Interleukin-13 and tumour necrosis factor-alpha synergistically induce eotaxin production in human nasal fibroblasts. AB - BACKGROUND: There is increasing evidence that eotaxin is a key mediator in the development of tissue eosinophilia. However, the mechanism involved in the production of eotaxin has yet to be clarified. Most recently, it has been shown that interleukin (IL) -4 induces eotaxin in dermal fibroblasts. A novel cytokine termed IL-13, which binds to the alpha-chain of the IL-4 receptor, shares many biological activities with IL-4. It is known that fibroblasts express the IL-4 receptor and produce collagen type I upon stimulation with IL-4. OBJECTIVE: We investigated whether IL-13, as well as IL-4, are able to induce eotaxin production in human nasal mucosal fibroblasts (HNMFs). Furthermore, we investigated the effect of costimulation of IL-13 and TNFalpha on eotaxin production. METHODS: HNMFs, isolated from inferior nasal mucosa samples, were stimulated by various kind of cytokines for 1-36 h at 37 degrees C in 5% CO2. The change in the expression of eotaxin mRNA was then evaluated by reverse transcriptase-polymerase chain reaction and the Southern blot analysis. The amount of eotaxin in the culture media was measured by ELISA. RESULTS: IL-13 as well as IL-4 dose-dependently induced eotaxin expression in HNMFs. Furthermore, IL-13 and TNFalpha synergistically induced eotaxin expression in HNMFs, while they hardly induced eotaxin expression in endothelial cells, epithelial cells or eosinophils. The synergy was observed when pre-incubation of HNMFs with IL-13 was followed by a stimulation with TNFalpha, or HNMFs were simultaneously stimulated with IL-13 and TNFalpha. CONCLUSION: These results strongly indicate that IL-13, as well as IL-4, may be important in eotaxin-mediated eosinophilic inflammation in nasal mucosa. In addition, in nasal mucosa, fibroblasts are the major cell source for eotaxin. PMID- 10691893 TI - Total and specific IgE (house dust mite and intestinal helminths) in asthmatics and controls from Gondar, Ethiopia. AB - BACKGROUND: The role, if any, of parasitosis in the development of asthma remains incompletely understood; both 'protective' and 'predictive' associations have been reported. We report a study which examined immunoglobulin (Ig) E responses to two common helminths in asthmatics living in Ethiopia. OBJECTIVE: To compare the frequencies of specific IgE antibodies to Ascaris and Necator species and to Der p 1 among 84 adult asthmatics and a referent group of nonasthmatics. METHODS: A case-control analysis. RESULTS: Total IgE levels were not different between the two groups. The presence of specific IgE to Der p 1 was strongly associated with asthma (P = 0.001). Raised levels of Ascaris-(P = 0.010) and Necator- (P = 0.001) specific IgE antibodies were more common among referents; there were no associations between specific IgE production to Der p 1 and either of the two parasites. CONCLUSION: These findings confirm the association between Der p 1 sensitization and asthma among urban, adult Ethiopians. While they also indicate a negative relationship with two indicators of helminth infestation it appears that this is not mediated through the immunological response to common aeroallergens. PMID- 10691894 TI - Detection of immunoglobulin antibodies in the sera of patients using purified latex allergens. AB - BACKGROUND: Latex allergy is largely an occupational allergy due to sensitization to natural rubber latex allergens present in a number of health care and household products. Although several purified allergens are currently available for study, information on the usefulness of these purified, native or recombinant allergens in the demonstration of specific immunoglobulin (Ig) E in the sera of patients is lacking. OBJECTIVE: To evaluate the purified latex allergens and to demonstrate specific IgE antibody in the sera of health care workers and spina bifida patients with clinical latex allergy. METHODS: Two radioallergosorbent and an enzyme-linked immunosorbent assay (ELISA) using latex proteins Hev b 1, 2, 3, 4, 6 and 7 along with two glove extracts and Malaysian nonammoniated latex (MNA) were evaluated to demonstrate IgE in the sera of health care workers and spina bifida with latex allergy and controls with no history of latex allergy. RESULTS: ELISA using the purified latex allergens demonstrated specific IgE in 32-65% health care workers and 54-100% of spina bifida patients with latex allergy. The corresponding figures for RAST were 13-48 and 23-85 for RAST-1 and 19-61 and 36 57 for RAST-2. These results were comparable with the results obtained with glove extracts and crude rubber latex proteins. CONCLUSIONS: When used simultaneously, latex proteins Hev b 2 and Hev b 7 reacted significantly with specific serum IgE in 80% of health care workers and 92% of spina bifida patients with latex allergy by ELISA technique, while this combination gave lower positivity when the RASTs were used. By the addition of Hev b 3, specific IgE was detected in all spina bifida patients with latex allergy. Both RASTs failed to show specific IgE in the control subjects, while the ELISA showed significant latex-specific IgE in 22% of controls. PMID- 10691896 TI - Indoor determinants of Der p 1 and Der f 1 concentrations in house dust are different. AB - BACKGROUND: Exposure to mite allergens is a major risk factor for sensitization and the development of asthma. Der p 1 and Der f 1 content in homes and probably the proportion of both antigens is highly variable even in the same geographical area. OBJECTIVE: We investigated specific indoor determinants of Der p 1 and Der f 1 concentrations in house dust of two German cities, Erfurt and Hamburg (n = 405 homes). METHODS: Mite allergen levels were determined using monoclonal antibodies against Der p 1 and Der f 1 by the ELISA method. Indoor relative humidity and temperature were monitored continuously in the homes over 1 week. The characteristics of homes and occupants were assessed by questionnaire to obtain information on factors which may have an impact on the mite antigen concentration in house dust. These determinants were studied by multivariate regression analysis. RESULTS: The correlation between concentrations of Der p 1 and Der f 1 inside the homes was weak (r = 0.29-0.35), indicating that different determinants are relevant. Concentrations of the allergens were significantly higher on lower floors (ratios 2-8 times, Der p 1, Der f 1), on old mattresses (ratios 3-13 times, Der p 1, Der f 1), in post-war buildings (ratio 6 times, Der p 1), for non-central heating (ratio 2 times, Der p 1), for old carpets (ratio 3 times, Der p 1) and for the presence of a dog in the house (ratio 3 times, Der f 1). Furthermore, mite concentration increases with raising relative humidity (ratio 1.03 per 1%, Der p 1) and with decreasing temperature (ratio 0.86 per 1 degrees C, Der p 1) indoors. CONCLUSION: Both Der p 1 and Der f 1 concentrations should be measured in house dust, since they are only weakly correlated and have different determinants. PMID- 10691895 TI - Sensitization to oilseed rape is not due to cross-reactivity with grass pollen. AB - BACKGROUND: Oilseed rape is an important crop grown in the UK which can cause specific immunological sensitization with clinical symptoms in a relatively small number of the general population. Individuals with immunoglobulin (Ig) E-mediated allergy to oilseed rape have also been found to be sensitized to other pollen allergens, most frequently being grass pollen. Cross-reactivity between common grass and oilseed rape would have important implications, especially as their flowering period coincides. OBJECTIVE: We have investigated whether the cosensitization found in individuals sensitized to both oilseed rape and grass pollen is due to cross-reactivity. METHODS: Cross-reactivity between oilseed rape and grass pollen was determined using RAST, RAST inhibition, Western blotting and inhibition studies with Western blotting. RESULTS: Competitive RAST inhibition studies between pollen of oilseed rape and grass failed to show any cross reactivity between the pollen types. Self-inhibition with oilseed rape resulted in 90% inhibition, whereas there was less than 10% inhibition with grass pollen. Western blotting revealed allergens of similar molecular weight in both oilseed rape and grass pollen. Despite allergens of similar molecular weights being present in both pollen types, inhibition immunoblot studies confirmed that the allergens in the two allergens were immunologically distinct. CONCLUSION: The allergens of oilseed rape and grass pollen, although similar in molecular weights, are immunologically distinct and there is no evidence of cross reactivity between them. Individuals allergic to grass pollen will not necessarily develop a specific nasal or airway response to inhaled oilseed rape pollens. PMID- 10691897 TI - The role of costimulatory molecules (B7-1 and B7-2) on allergen-stimulated B cells in cedar pollinosis subjects. AB - BACKGROUND: B7-1 (CD80) and B7-2 (CD86), which are costimulatory molecules in T cell activation, play important roles in the differentiation of TH1- or TH2 phenotypes. These molecules were also suggested to play important roles in sensitization to a cedar pollen antigen by blocking studies using neutralizing antibodies, but there have been very few studies concerning the effects following induction by antigen. OBJECTIVE: In this study, we investigated the roles of B7-1 and B7-2 in the differentiation of TH1 and TH2 subsets after stimulation with the antigen in subjects with cedar pollinosis. METHODS: Skin-prick test was performed in nine subjects with pollinosis and in nine normal controls. Peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with Japanese cedar pollen extract. After in vitro stimulation, the expression of CD80 and CD86 on CD19+ cells was analysed by two-colour flow cytometry. Culture supernatants were collected for all subjects and the production of type 1 and type 2 cytokines was measured by ELISA. RESULTS: After in vitro stimulation, the expression of CD80 (B7-1) was upregulated in both pollinosis and control subjects, but no significant difference was observed between the two groups. On the other hand, CD86 (B7-2) was significantly upregulated following stimulation in pollinosis subjects (P = 0.02). A significantly higher level of IL-5 (P = 0.04) was produced by PBMCs of pollinosis subjects than by those of normal controls. A significantly higher level of interferon (IFN)-gamma (P = 0.03) was produced by PBMCs of normal controls than by those of pollinosis subjects. CONCLUSION: These results indicated that TH2 response was predominant in pollinosis subjects, and that CD19+ cells of pollinosis subjects expressed higher levels of B7-2 than those of control subjects after in vitro stimulation. In pollinosis subjects, B7-2 rather than B7-1 may be the costimulatory molecule involved in allergen-induced activation of PBMCs. PMID- 10691898 TI - Family patterns of asthma, atopy and airway hyperresponsiveness: an epidemiological study. AB - BACKGROUND: The patterns of inheritance of asthma have largely been explored using data of symptom history collected by questionnaires which are subject to bias and which may therefore distort the measured relationship. OBJECTIVE: The purpose of this study was to examine family patterns of allergic disease using objective measurements of atopy and of airway hyperresponsiveness (AHR). METHODS: A large random sample of children aged 8-11 years was studied and 3 months later, their parents were also invited for study. Of the sample of 1655 children, both parents of 661 children were studied. In all subjects, respiratory illness history was measured by questionnaire, atopy by skin tests and AHR by responsiveness to histamine. RESULTS: The odds ratio for a child to have AHR if either parent had the same condition was approximately 2. 0, which was the same as the odds ratio for wheeze or diagnosed asthma in the presence of the same condition in either parent. The odds ratio for atopy was smaller (approximately 1.4, NS) but the risk of a nonatopic child having AHR if the parent had AHR was 3.0 (P = 0.01). The correlation between weal size in the child and parent was poor and the severity of AHR in the child was only modestly correlated with the severity of AHR in the parent (R = 0.51, P = 0.04). CONCLUSION: The use of objective measurements did not strengthen the association between atopic or asthmatic conditions in the parent and child, but did suggest that atopy and AHR are inherited independently. PMID- 10691899 TI - Infant feeding patterns affect the subsequent immunological features in cow's milk allergy. AB - BACKGROUND: The first exposure to food antigens provokes an immune reaction in an infant, its type depending on the quantity and frequency of doses and the age at introduction, and also being influenced by genetic factors. Most infants develop tolerance to food antigens, but in a small minority they provoke adverse symptoms. OBJECTIVE: To study the effects of breast and formula feeding and other environmental and genetic factors on the subsequent type of cow's milk allergy classified by the presence or absence of immunoglobulin (Ig) E antibodies to cow's milk. METHODS: A cohort of 6209 infants was followed prospectively from birth for symptoms of cow's milk allergy. The infant-feeding regimen was recorded at the maternity hospital and at home. At a mean age of 6.7 months, a total of 118 infants (1.9%) reacted adversely to a challenge with cow's milk. Before the challenge, the response to a skin-prick test with cow's milk and serum IgE cow's milk antibodies was measured. RESULTS: At challenge, 75 (64%) infants showed IgE positive reactions to cow's milk, their most common symptom being acute-onset urticaria. Significant risk factors for the presence of IgE cow's milk antibodies in allergic infants were long breast-feeding (odds ratio [OR] 3.9, 95% confidence interval [CI] 1.6-9.8), exposure to cow's milk at the maternity hospital (OR 3.5, 95% CI 1.2-10.1) and breast-feeding during the first 2 months at home either exclusively (OR 5.1, 95% CI 1.6-16.4) or combined with infrequent exposure to small amounts of cow's milk (OR 5.7, 95% CI 1.5-21.6). Fifty infants had their first adverse symptoms during exclusive breast-feeding, and 32 infants were sensitized during exclusive breast-feeding. Most of the infants in both cases were IgE-positive: 37 and 23, respectively. CONCLUSIONS: In infants who are prone to developing cow's milk allergy, prolonged breast-feeding exclusively or combined with infrequent exposure to small amounts of cow's milk during the first 2 months of life induces development of IgE-mediated response to cow's milk. PMID- 10691900 TI - Clinical relevance of food additives in adult patients with atopic dermatitis. AB - BACKGROUND: Adverse reactions to food play an important role in the pathogenesis of atopic dermatitis (AD). In infancy and childhood, food allergies are observed in up to 30%, whereas nonallergic hypersensitivity reactions (pseudoallergic reactions) towards food additives have been reported to occur between 2 and 7%. By contrast, sensitizations towards food allergens are rarely of clinical relevance in adults and little data is available on nonallergic hypersensitivity reactions. To date the role of pseudoallergic reactions as an aggravating factor in AD of adult patients remains controversial. However, many adult patients report on food-related aggravation of the disease and nonallergic hypersensitivity reactions have been incriminated repeatedly. OBJECTIVE: To elucidate the relevance of food additives in adult patients suffering from AD. METHODS: Fifty patients were monitored over 4 weeks under regular diet followed by 6 weeks of a diet omitting known pseudoallergens. Skin status of patients was assessed every 2 weeks by a standardized scoring, and serum eosinophilic cationic protein (ECP) was determined before and after diet. RESULTS: Nine of fifty patients dropped out, 26 showed a significant improvement of the Costa-score by 57%. In 23/26 patients a corresponding reduction of serum ECP level by 52% on average was determined. Responder patients (24/26) were orally challenged with food rich in pseudoallergens followed by double-blind exposure to food additives (n = 15). A worsening of the eczema was seen in 19/24 patients after intake of pseudoallergen-rich food and in 6/15 patients after exposure to food additives. CONCLUSION: These results indicate that a subgroup of adult patients with AD clinically improve on low-pseudoallergen diet but only a small subgroup respond to oral provocation with food additives. PMID- 10691901 TI - Late asthmatic reaction with airway inflammation but without airway hyperresponsiveness. PMID- 10691902 TI - Window pane condensation and high indoor vapour contribution - markers of an unhealthy indoor climate? AB - OBJECTIVE: The aim of this study was to investigate whether window pane condensation and indoor vapour contribution >/= 3 g/m3 could be used as indicators of defective air change rate, high indoor humidity and high mite allergen concentration in mattress dust. METHODS: Actual ventilation rate, indoor temperature, air humidity (AIH/RH) and concentrations of mite allergen were measured in 59 houses and compared with received outdoor temperatures and air humidity. Indoor vapour contribution defined as the difference between the indoor and the outdoor vapour concentration was calculated. Sensitivity, specificity, predictive values and accuracy were calculated for window pane condensation and high vapour contribution (>/= 3 g/m3), as indicators of defective ventilation (< 0.5 ACH), high indoor humidity (>/= 7 g/kg and >/= 45% RH) and high mite allergen concentration in mattress dust (>/= 2 microg/g). RESULTS: All houses with high humidity and high mite allergen concentrations were positive for the two indicators (high sensitivity), but with a specificity of only 50% so that half of the houses with reported condensation and high vapour contribution turned out to be low pollution houses with adequate high ventilation levels. Both indicators had high negative predictive values and absence of the two indicators almost certainly (97-100%) excluded high indoor pollution with high humidity and high mite concentrations. Overall more than 70% of the dwellings were correctly classified by the two indicators. CONCLUSION: Absence of window pane condensation on double-glazed windows and low indoor vapour contribution (< 3 g/m3) during the winter are true markers of a dwelling without high indoor air humidity and without high mite allergen concentrations in mattress dust in houses in a cold temperate climate with subzero outdoor temperatures. The presence of the two indicators is associated with a 18-45% risk of high humidity and mite allergen concentrations so in this latter group further measurements are needed for correct classification. PMID- 10691903 TI - Airborne endotoxin exposure and the development of airway antigen-specific allergic responses. AB - BACKGROUND AND OBJECTIVE: Repeated exposure of aerosolized antigen via respiratory tract can induce immunoglobulin (Ig) E isotype-specific tolerance to this antigen. However, the atopic individuals often produce a higher titre of IgE in response to airborne environmental allergens. The mechanisms of this differential regulation of airway allergen-specific immune responses are not fully understood. This study investigated the role of airborne endotoxin on the initiation of antigen-specific airway allergic responses. METHODS: ELISA methods for detection of isotypes of antigen-specific antibodies and competitive reverse transcription polymerase chain reaction for detection mRNA of cytokines were used. In addition, Liu stain method was used to analyse the amounts of eosinophils in bronchoalveolar lavage fluid. RESULTS: Mice pre-exposed with airborne endotoxin mounted significantly higher amounts of OVA-specific IgE antibody responses to inhaled OVA than those OVA-only sensitized mice. Inhaled endotoxin could downregulate repeated airway antigen exposure-induced IgE isotype specific tolerance and increase antigen-induced lung eosinophils infiltration. CONCLUSIONS: These data show that airborne endotoxin exposure could potentiate allergen-specific airway inflammation. The results should have potential implications for understanding the development of allergen-induced airway allergic responses. PMID- 10691904 TI - Altered in vitro apoptosis of cultured mast cells prepared from an inbred strain of mice, NC/Kuj. AB - BACKGROUND: An inbred strain of mice, NC, develops dermatitis associated with highly elevated serum IgE and dermal mast cell hyperplasia. OBJECTIVES AND METHODS: To clarify the mechanisms for the dermal mast cell hyperplasia in NC, we prepared bone marrow-derived mast cells (BMMCs) from three strains of mice, NC/Kuj, C57BL/6 and BALB/c, and compare histamine contents, histamine release abilities, adhesive properties and apoptosis of the BMMCs. RESULTS: Compared with BMMCs obtained from C57BL/6 and BALB/c, NC/Kuj BMMC possessed higher histamine content and higher adhesive ability to plastic plates, although histamine release from BMMCs was found to be similar in the three strains. The most intriguing finding is the lack of apoptosis in the BMMCs from NC/Kuj upon growth factor deprivation as determined by DNA ladder formation and loss of membrane phospholipid asymmetry. CONCLUSION: The altered in vitro properties of mast cells in NC/Kuj partially account for an increase of dermal mast cells, which might be involved in the development of skin lesions in NC. PMID- 10691905 TI - Bronchoprotective effects of atrial natriuretic peptide against propranolol induced bronchoconstriction after allergic reaction in guinea pigs. AB - OBJECTIVE: To study the effects of intravenous atrial natriuretic peptide (ANP) on antigen-induced bronchoconstriction, propranolol-induced bronchoconstriction (PIB) after antigen challenge, and histamine-induced bronchoconstriction in guinea pigs. METHODS: Allergic bronchoconstriction was evoked by inhalation of ovalbumin (OA) and PIB was caused when 10 mg/mL of propranolol was inhaled 20 min after OA challenge in passively sensitized and artificially ventilated guinea pigs. 25, 50, 100 and 200 microg/mL of histamine were inhaled for 20 s at 5-min intervals in non-sensitized guinea pigs. RESULTS: Pretreatment with ANP in doses of 0.1 and 1.0 nmol/kg injected intravenously 15 min after antigen challenge reduced PIB in a dose-dependent manner, and 5 min before antigen challenge significantly attenuated PIB but not antigen-induced bronchoconstriction. Intravenous ANP significantly reduced bronchial responses to increasing concentrations of inhaled histamine in a dose-dependent manner. CONCLUSION: These results suggest that ANP possesses protective effects against propranolol-induced and histamine-induced bronchoconstriction, albeit by a non-specific mechanism in guinea pig in vivo. PMID- 10691906 TI - Possible mechanism of bronchoprotection by SIN-1 in anaesthetized guinea pigs: roles of nitric oxide and peroxynitrite. AB - BACKGROUND: S-morpholinosydnonimine (SIN-1) is thought to generate peroxynitrite. Recent reports suggested that peroxynitrite possessed a potent vascular relaxant activity via guanylate cyclase activation. However, no previous studies have examined the relaxant effect of peroxynitrite on airway smooth muscle. OBJECTIVE: To determine the mechanism of bronchoprotection by SIN-1, considering in particular the involvement of nitric oxide (NO) and peroxynitrite. METHODS: Peroxynitrite formation was assayed by monitoring the oxidizing activity of dihydrorhodamine 123, and NO was measured polarographically as a redox current in vitro. We examined the effect of SIN-1 delivered to the airway by ultrasonic nebulization against bronchoconstriction induced by acetylcholine in anaesthetized guinea pigs. RESULTS: SIN-1 produced peroxynitrite in a time- and concentration-dependent manner, but did not produce NO in vitro. However, when mixed with glutathione (GSH) and bronchoalveolar lavage fluid (BALF), peroxynitrite formation by SIN-1 was inhibited and SIN-1 induced the release of NO. SNAP (S-nitroso-N-acetyl-penicillamine) and SIN-1 each inhibited acetylcholine-induced bronchoconstriction in a dose-dependent manner in vivo. Though GSH alone did not have any effect on baseline airway resistance and acetylcholine-induced bronchoconstriction, pretreatment with GSH significantly enhanced SNAP- and SIN-1-induced bronchoprotection. In addition, pretreatment with carboxy-PTIO, a NO scavenger, completely inhibited bronchoprotective effect of SNAP on acetylcholine-induced bronchoconstriction, but partially inhibited SIN 1-induced bronchoprotection. CONCLUSION: These findings demonstrated that SIN-1 is a potent peroxynitrite-releasing compound and caused significant bronchoprotection against acetylcholine. The mechanism of bronchoprotection by SIN-1 appears to be mediated by peroxynitrite but also at least in part through NO regeneration, which may involve GSH and airway thiols as a consequence of exposure to peroxynitrite. PMID- 10691907 TI - Early B cell defects. PMID- 10691908 TI - Clonal expansion of T cells infiltrating in the airways of non-atopic asthmatics. AB - CD4+ T cells are thought to play an important role in airway inflammation in both atopic and non-atopic asthma. However, the mechanism by which T cells are activated in non-atopic asthma, where there is no causative antigen identified, is unknown. To elucidate this issue, we analysed T cell receptor (TCR) Vbeta gene clonotypes of T cells in the bronchoalveolar lavage fluids (BALF) of non-atopic asthmatics using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis and a sequencing method. We found that the numbers of TCR Vbeta gene clonotypes of T cells in the BALF of non-atopic asthmatics were significantly increased compared with those of peripheral blood lymphocytes (PBL). We also found that there were several shared amino acid motifs in complementarity-determining region 3 (CDR3) of TCR Vbeta genes from those T cell clones in BALF of non-atopic asthmatics, whereas these shared motifs were not found in the same Vbeta family genes from PBL in the patients. Moreover, a conserved amino acid sequence was detected in two patients who shared a common HLA-DR allele. These results indicate that the infiltrating T cells in the airways of non-atopic asthmatics recognize relatively limited epitopes of antigens and are clonally expanded by antigen-driven stimulation. PMID- 10691909 TI - Targeted Fc2'-3-PE40 chimeric protein abolishes passive cutaneous anaphylaxis in mice. AB - The alarming increase in the incidence of allergic diseases in the past decade has led to a clear call for more effective treatment. Recently, we reported on the construction of a chimeric protein for targeted elimination of cells expressing FcepsilonRI receptors. This chimeric protein, designated Fc2'-3-PE40, is composed of a Fc fragment of mouse IgE attached to a truncated form of Pseudomonas exotoxin. The Fc2'-3-PE40 chimeric protein was found to be highly cytotoxic to mouse mast cell lines and primary mouse mast cells. We now demonstrate that Fc2'-3-PE40 successfully prevents the development of passive cutaneous anaphylaxis reaction (PCA) in mice. Treatment with Fc2'-3-PE40 for 7 days prevented the PCA reaction in mice by 80% compared with that in control mice given only PBS. Fc2'-3-PE40M, the mutated, enzymatically inactive analogue of Fc2'-3-PE40, did not display this activity. Fc2'-3-PE40 was also effective when given as a single dose 16 h before antigen exposure, resulting in complete inhibition of the PCA reaction. Moreover, treatment with Fc2'-3-PE40 did not cause mast cell degranulation, as the serum histamine values of mice treated with Fc2'-3-PE40 were within the range obtained for control, untreated mice. Thus, the Fc2'-3-PE40 chimeric protein offers a novel approach to the treatment of allergic disorders. PMID- 10691910 TI - CD4-CD8- T cells bearing invariant Valpha24JalphaQ TCR alpha-chain are decreased in patients with atopic diseases. AB - Atopic disorders are caused by disregulated activation of T helper 2 (Th2) cells that produce IL-4 and IL-5. Because the presence of IL-4 potently augments the differentiation of naive T cells into Th2 cells, it is important to seek the cell population which provides IL-4 for naive T cells. Recently, a unique subpopulation of T cells, natural killer (NK) T cells, has been shown to produce a large amount of IL-4 upon activation, suggesting their regulatory role in initiation of Th2 cell differentiation. To determine whether NK T cells play a regulatory role in human Th2 cell-mediated atopic diseases, we analysed the frequency of invariant Valpha24JalphaQ CD4-CD8- double-negative (DN) T cells, human NK T cells, in patients with atopic asthma and atopic dermatitis. We also studied cytokine production from Valpha24+ Vbeta11+ DN T cells, which comprise most of Valpha24JalphaQ DN T cells. We found that the invariant Valpha24JalphaQ DN T cells were greatly diminished in patients with asthma and atopic dermatitis. On the other hand, there was no significant difference in Valpha24+ CD4+ T cells possessing invariant Valpha24JalphaQ TCR between healthy subjects and atopic patients. We also found that Valpha24+ Vbeta11+ DN T cells from healthy subjects predominantly produced interferon-gamma (IFN-gamma) but not IL-4 upon activation. These results suggest that NK T cells may not be essential for human atopic disease and that the disappearance of NK T cells, most of which produce IFN gamma, may be involved in the pathogenesis of atopic diseases. PMID- 10691911 TI - Interferon-alpha (IFN-alpha) stimulates anti-melanoma cytotoxic T lymphocyte (CTL) generation in mixed lymphocyte tumour cultures (MLTC). AB - IFN-alpha administration after primary tumour resection improves the survival of melanoma patients at high risk of relapse. To investigate whether this response might be due to stimulation of anti-tumour immunity, the effect of IFN-alpha on anti-melanoma CTL generation in MLTC was measured. IFN-alpha increased both allogeneic and autologous anti-melanoma CTL generation from peripheral blood lymphocytes stimulated with irradiated primary melanoma cultures. IFN-alpha up regulated MHC class I expression on primary melanoma cultures, whereas IFN-gamma up-regulated both MHC class I and II expression. However, the effect of IFN-alpha on anti-melanoma CTL generation was often more potent than that of IFN-gamma, equalling the effect of the optimal combination of IL-2 and IL-12. Pre-treatment of primary melanoma cultures with IFN-gamma was sufficient for CTL generation in MLTC, whereas IFN-alpha needed to be present during the MLTC. While direct anti proliferative effects of IFN-alpha on some tumour cells have been described, IFN alpha did not inhibit proliferation of primary melanoma cultures. These results suggest that the clinical effects of IFN-alpha in melanoma patients may be immune mediated. PMID- 10691912 TI - Autoimmunity to glutamic acid decarboxylase in patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). AB - Antibodies to glutamic acid decarboxylase (GAD) occur frequently in patients with APECED, although clinical insulin-dependent diabetes mellitus (IDDM) is seen only in a subgroup of the patients. We studied the cellular immunity to GAD, antibodies to GAD and their association with the HLA DQB1 risk alleles for IDDM in patients with APECED. Proliferation responses to GAD were enhanced in the patients with APECED when compared with the control subjects (P = 0.004), but autoimmunity to GAD was not associated with IDDM in APECED. The levels of interferon-gamma (IFN-gamma) secreted by GAD-stimulated T cells were higher in the patients than in control subjects (P = 0. 001). A negative correlation (r = - 0.436, P = 0.03) existed between the antibody levels and the stimulation indices (SIs) to GAD. In 14 non-diabetic patients no difference in insulin secretion was observed in intravenous glucose tolerance test (IVGTT) between the patients with and without T cell reactivity to GAD. We conclude that cellular immunity to GAD detected as T cell proliferation response to GAD or IFN-gamma secretion by GAD stimulated T cells was frequent in patients with APECED (69%) and was not restricted to the patients with clinically detectable beta-cell damage. PMID- 10691913 TI - IL-10 enhances IL-2-induced proliferation and cytotoxicity by human intestinal lymphocytes. AB - IL-10 modulation of human intestinal T lymphocyte functions was studied for the first time. Lymphocyte proliferation was determined by 3H-thymidine incorporation; cytokine production, by ELISA; expression of surface markers, by immunofluorescence and flow cytometric analysis; and cytotoxicity, by lysis of 51Cr-labelled target cells. IL-10 blocked phytohaemagglutinin (PHA)-induced activation and proliferation of CD8+ T cells from the epithelium and lamina propria. It was a greater inhibitor of IL-2, interferon-gamma, and tumour necrosis factor-alpha production than were IL-4 or transforming growth factor beta. In contrast, IL-10 enhanced IL-2-stimulated proliferation of both CD4+ and CD8+ T cells by increasing cell division after activation. It also augmented IL-2 but not IL-15-induced cytotoxicity of intestinal lymphocytes against colon cancer by a mechanism independent of natural killer cells. In conclusion, IL-10 blocking of proinflammatory cytokine secretion probably reduces intestinal inflammation. IL-10 augmentation of IL-2-induced cytotoxicity may help to maintain host defence. PMID- 10691914 TI - Proliferative responses of blood mononuclear cells (BMNC) in a cohort of elderly humans: role of lymphocyte phenotype and cytokine production. AB - Age-related impaired T cell function is associated with increased mortality risk. The purpose of the present study was therefore to identify factors associated with the age-related decreased phytohaemagglutinin (PHA)-induced proliferative response of lymphocytes in a cohort of 174 81-year-old humans and in 91 young controls. Decreased proliferation was associated with a reduced number of true naive CD4+ cells (CD62L+CD45RO-). Furthermore, a low IL-2-stimulated proliferation was correlated with a decreased PHA response in the elderly cohort, whereas reciprocal interactions of IL-10- and IL-2-producing cells were of importance in both elderly and young subjects. Accordingly, a minimum of true naive CD4+ cells was required for a normal proliferative response to PHA, perhaps by providing sufficient IL-2 which is critical for growth of naive as well as memory cells. PMID- 10691915 TI - Prednisolone inhibits cytokine-induced adhesive and cytotoxic interactions between endothelial cells and neutrophils in vitro. AB - We assessed whether prednisolone influenced the ability of human polymorphonuclear neutrophils (PMN) to adhere to and cause lysis of human umbilical vein endothelial cells (HUVEC) in vitro (as measured by the release of 51Cr). Pretreatment of the endothelium with IL-1beta or tumour necrosis factor alpha (TNF-alpha) caused prominent endothelial E-selectin expression and endothelial hyperadhesiveness for neutrophils, as well as PMN-mediated cytotoxicity. All these processes were dose-dependently reduced when prednisolone was added to the assay system. This protective effect remained when HUVEC alone were pretreated with the drug prior to washing and cytokine activation. Likewise, when HUVEC cytotoxicity was induced by the nitric oxide (NO) donor S-nitroso acetyl-penicillamine (SNAP), prednisolone reduced cell injury significantly. In contrast, prednisolone did not interfere with signalling systems between TNF alpha-stimulated HUVEC and quiescent PMN such as IL-8 generation and release of cytosolic Ca2 + in the PMN. Thus, in this in vitro model of vasculitis, prednisolone dose-dependently reduced cytokine-induced E-selectin expression and HUVEC hyperadhesiveness for neutrophils, as well as reducing neutrophil-dependent cytotoxicity against HUVEC via NO-dependent steps. PMID- 10691916 TI - PPD-specific IgG1 antibody subclass upregulate tumour necrosis factor expression in PPD-stimulated monocytes: possible link with disease pathogenesis in tuberculosis. AB - Cachexia is a prominent feature of advanced tuberculosis, in association with increased expression of the monokine tumour necrosis factor (TNF)-alpha. Monocytes, have high affinity receptors (mannose, complement and Fc gamma1 and gamma111) which mediate antigen uptake and subsequent cytokine activation. Several mycobacterial proteins, including PPD, can stimulate TNF-alpha secretion from monocytes. However, the role of various receptors in stimulating or regulating TNF-alpha secretion is still unclear. We have previously shown selective augmentation of opsonic antibodies (IgG1 and IgG3) in tuberculosis patients with advanced pulmonary disease. We now analyse the role of opsonizing antibodies in modulating TNF-alpha expression in antigen stimulated monocytes. PPD was used as the prototypic mycobacterial antigen to stimulate monocytes from PPD skin test negative donors (n = 7) in the presence of plasma from tuberculosis patients (n = 8), containing known amounts of IgG1 and IgG3 anti-PPD antibodies. TNF-alpha secretion was enhanced in the presence of TB plasma (4/8) but not in the presence of control plasma. Using Spearman Rank analysis (two-tailed Fisher exact test), a significant correlation (rho = 0.762; P = 0. 04) was observed between IgG1 antibodies and enhancement of TNF-alpha secretion. No significant association was observed with IgG2 (rho = 0.310; P = 0.41), IgG3 (rho = 0.089; P = 0.81) or IgG4 (rho = - 0.357; P = 0.347) subclass antibodies. Absorption of IgG1 with protein 'A' removed the enhancement of TNF-alpha secretion activity from the plasma samples. Our results therefore indicate that IgG1 antibodies may enhance the chronic release of TNF-alpha in TB patients with progressive disease and, for the first time, show a direct link between disease pathogenesis and raised antibody levels. PMID- 10691917 TI - Bacteroides fragilis enterotoxin induces the expression of IL-8 and transforming growth factor-beta (TGF-beta) by human colonic epithelial cells. AB - Bacteroides fragilis toxin (BFT) has been shown to be capable of inducing intestinal mucosal inflammation in animals. Such inflammation may be responsible for diarrhoea, which occurs in some, but not all human carriers of enterotoxigenic strains of B. fragilis (ETBF). We have studied responses to BFT by different human intestinal epithelial cell lines and subsequently investigated the expression of IL-8 and TGF-beta by T84 cells. The latter were selected because their responses to BFT, characterized by morphological changes and cell death by apoptosis, were similar to those we have recently observed in primary human colonocytes. We show that BFT dose-dependently increased the expression of transcripts and protein of the polymorphonuclear cell chemoattractant IL-8. BFT also dose-dependently induced the release of TGF-beta, which has been shown to enhance the repair of the injured intestinal epithelium. However, the secreted TGF-beta was almost exclusively in the biologically inactive form, as determined by Mv1Lu bioassay. Our studies therefore suggest that exposure of colonic epithelial cells in vivo to high concentrations of BFT can initiate an inflammatory response via secreted IL-8. BFT-induced release of latent TGF-beta may facilitate the subsequent repair of the injured epithelium, following its activation by proteases from neighbouring cells. Variation in cytokine responses by colonic epithelial cells in vivo could be an important determinant in the development of mucosal disease and symptoms in response to ETBF. PMID- 10691918 TI - Expression of HLA-DR, costimulatory molecules B7-1, B7-2, intercellular adhesion molecule-1 (ICAM-1) and Fas ligand (FasL) on gastric epithelial cells in Helicobacter pylori gastritis; influence of H. pylori eradication. AB - There is evidence that Helicobacter pylori infection up-regulates the expression of HLA class II molecules by gastric epithelial cells (GEC). In this study we evaluated whether GEC are capable of expression of costimulatory molecules in H. pylori gastritis. The expression of FasL by GEC, before and after eradication of H. pylori, was also studied. Thirty patients (23 men) aged 27-81 years (53.67 +/- 13.99 years (mean +/- s.d.)) with dyspepsia were studied. Upper gastrointestinal endoscopy was performed and six biopsies were obtained (antrum, n = 3; corpus, n = 3) for Campylobacter-Like Organisms (CLO) test and histology; 23 (16 men) were H. pylori+ and seven (all men) were H. pylori- by both methods and served as controls. Helicobacter pylori eradication therapy was given to H. pylori+ patients and all patients were re-endoscoped after 116 +/- 9 days. Formalin-fixed paraffin-embedded tissue sections were stained by the ABC immunoalkaline phosphatase method. In H. pylori gastritis HLA-DR was expressed and correlated with disease activity (P < 0.01). No HLA-DR was observed in controls. In H. pylori-eradicated patients significant decrease of HLA-DR was found (antrum, P < 0. 001). ICAM-1 was expressed by GEC in 80% of H. pylori+ patients; ICAM-1 expression did not correlate with gastritis parameters and decreased significantly after eradication (antrum, P < 0.01). B7-1 and B7-2 were expressed on H. pylori+ samples and their expression decreased after eradication, albeit not significantly. Weak epithelial expression of both B7 molecules was observed in all the controls. FasL was steadily expressed by GEC in both H. pylori+ and H. pylori- patients and remained almost unchanged after eradication. These findings suggest that GEC may acquire antigen-presenting cell properties in H. pylori infection through de novo expression of HLA-DR and costimulatory molecules. This seems to be attenuated after eradication and resolution of mucosal inflammation. The same cells exhibit the capacity to control the inflammatory process, probably by inducing apoptotic cell death to Fas-bearing infiltrating lymphocytes. PMID- 10691919 TI - Mechanisms of the in vitro fungicidal effects of human neutrophils against Penicillium marneffei induced by granulocyte-macrophage colony-stimulating factor (GM-CSF). AB - We examined the in vitro fungicidal activity of human neutrophils against conidia and yeast cells of Penicillium marneffei. Neutrophils showed a small but significant anti-fungal effect against the yeast form of P. marneffei. Treatment of neutrophils with GM-CSF significantly augmented their anti-fungal activity. In contrast, the conidia form resisted killing even by stimulated neutrophils. Neutrophil fungicidal effect was not inhibited by superoxide dismutase (SOD), while the same treatment significantly suppressed the killing of Candida albicans by GM-CSF-stimulated neutrophils. For effective killing of P. marneffei yeast cells by GM-CSF-stimulated neutrophils, direct contact between the two was essential; interference in such interaction by separation using a 0. 45-microm pored membrane prevented such an effect. Addition of colchicine attenuated GM-CSF stimulated neutrophil fungicidal activity in a dose-dependent manner. This effect did not appear to be mediated by interference with neutrophil mobility toward yeast cells, because similar results were obtained when the cultures were set in round-bottomed wells which facilitate their direct contact. Finally, granular extracts derived from unstimulated neutrophils significantly suppressed the growth of microorganisms. Pretreatment of neutrophils with GM-CSF markedly enhanced this effect. The fungicidal activity of granular lysates was strongly, but not completely, reduced by heat treatment. Considered together, our results indicate that GM-CSF-stimulated neutrophils killed the yeast form of P. marneffei present in close proximity, probably in a superoxide anion-independent mechanism, but through exocytosis of granular enzymes which were largely heat-labile. PMID- 10691920 TI - Interferons and interferon (IFN)-inducible protein 10 during highly active anti retroviral therapy (HAART)-possible immunosuppressive role of IFN-alpha in HIV infection. AB - Interferons play an important, but incompletely understood role in HIV-related disease. We investigated the effect of HAART on plasma levels of IFN-alpha, IFN gamma, neopterin and interferon-inducible protein 10 (IP-10) in 41 HIV-infected patients during 78 weeks of therapy. At baseline HIV-infected patients had raised levels of both IP-10 and IFN-alpha compared with healthy controls (n = 19), with particularly high levels in advanced disease. HAART induced a marked decrease in levels of both IFN-alpha, neopterin and IP-10, though not to normal concentrations. In contrast, IFN-gamma levels were low throughout the study, and not different from controls. While neopterin and IP-10 remained significantly decreased compared with baseline levels throughout the study, IFN-alpha levels returned to baseline at the end of the study. Persistently high IP-10 and IFN alpha levels were associated with immunological treatment failure and even high baseline levels of IFN-alpha appeared to predict immunological relapse. Furthermore, we found a markedly suppressive effect of exogenously added IFN alpha on phytohaemagglutinin-stimulated lymphocyte proliferation in both patients and controls, and this suppressive effect seemed not to involve enhanced lymphocyte apoptosis. Our findings suggest a pathogenic role of IFN-alpha in HIV infection, which may be a potential target for immunomodulating therapy in combination with HAART. PMID- 10691921 TI - HIV-1 co-receptor expression on trophoblastic cells from early placentas and permissivity to infection by several HIV-1 primary isolates. AB - We examined CD4 and major HIV-1 co-receptor expression by trophoblast cells (TC) from early placentas, and the permissiveness of TC for infection by several natural HIV-1 isolates in vitro. Ten early placentas (4-6 weeks of gestation) from HIV- women were obtained after elective abortion. CD4 and HIV-1 co-receptor expression by TC was examined in terms of both mRNA and protein. The same TC were then challenged with three clinical HIV isolates of known phenotype, two originating from mothers who transmitted the virus to their child and one from a vertically infected newborn. TC infection was detected by polymerase chain reaction. CD4 expression was detected in five of the 10 placentas, while membrane protein expression of CCR3, CXCR4 and CCR5 was detected in every case, despite quantitative differences among individuals. Bonzo, GPR1 and ChemR23 mRNAs were detected in all TC preparations. TC from seven out of eight placentas were permissive to HIV entry, but no productive viral replication was detected (reverse transcriptase activity in culture supernatants). Interestingly, the addition of chemokine(s) or a CD4-blocking antibody to the cultures failed to inhibit TC virus entry. These data point to marked interindividual variability in HIV co-receptor expression by trophoblast cells and show that TC from early placentas can be infected in vitro by clinical HIV-1 isolates. They also suggest that viral entry in vitro might occur through a mechanism independent of both CD4 and chemokine receptors. PMID- 10691922 TI - CD4+ and CD8+ T lymphocyte regeneration after anti-retroviral therapy in HIV-1 infected children and adult patients. AB - Previous studies have shown a slow recovery of naive CD4+ T cell counts after anti-retroviral therapy in HIV-1-infected adults, which is in accordance with thymus atrophy after puberty. Here we investigate whether or not different patterns of naive CD4+ and CD8+ T cell repopulation are present in adult and child patients undergoing anti-retroviral treatment. Thus, 25 adults under highly active anti-retroviral therapy and 10 children under combined anti-retroviral therapy were retrospectively analysed for T cell subpopulations at baseline (T0) and around week 12 (T1) and week 24 (T2) of anti-retroviral treatment. Mean serum HIV-1 RNA levels dropped in both groups. Recovery of T cells in adults was characterized by a heterogeneous response between patients, with only 44% of them increasing their naive CD4+ and CD8+ T cell counts at T1, and changes in mean total CD4+ T cells were mainly shaped by memory cells. Otherwise, children were characterized by an early increase in naive T cells. Thus, at T1, all children analysed had a strong rise in CD4+ (from 389 +/- 116 to 569 +/- 121 cells/microl; P < 0.01), and nine out of 10 also in naive CD8+ T cells (from 244 +/- 58 to 473 +/- 85 cells/microl; P < 0.05). However, no significant correlation between age and naive repopulation was observed (P = 0. 22) in children. Thus, children had the earlier and greater increases in naive T cell subsets than adults, probably due to a more active thymus, with the potential for immune reconstitution when HIV-1 replication is controlled. PMID- 10691923 TI - CD28 costimulation and CD28 expression in T lymphocyte subsets in HIV-1 infection with and without progression to AIDS. AB - In a prospective study of 152 HIV-1 patients (with and without progression to AIDS) we examined CD28 MoAb costimulation and CD3 MoAb response using whole blood culture at baseline and up to either the time of AIDS diagnosis or the end of the observation period. CD28 antigen expression on both CD4+ and CD8+ T lymphocytes was also studied in both groups of patients. In patients who progressed to AIDS, CD28 MoAb costimulation was found to be decreased. Univariate time-dependent analysis showed that decreases in (i) absolute numbers of either CD4+, CD4+CD28+, CD8+CD28+ T cells, (ii) CD28 MoAb costimulation, and (iii) CD3 MoAb response, and an increase in CD8+CD28- %, are significant predictors for progression to AIDS. In addition, multivariate time-dependent analysis demonstrated that a decrease in CD28 MoAb costimulation (but not a decrease in CD3 MoAb response) was predictive for progression to AIDS, as were decreases in the percentage of CD4+ T cells and the absolute number of CD4+CD28+ T cells. Thus, CD28 MoAb costimulation can be considered a useful assay for monitoring HIV-1 infection. Furthermore, apart from the early increase in the percentage of CD8+CD28- T cells and an increase in the percentage of CD28- on CD8+ T cells in both groups of patients at baseline compared with normal controls, a negative correlation was found to exist between the percentages of CD4+ or CD4+CD28+ T cells and the percentage of CD8+CD28- T cells; this suggests that these cells are probably mutually regulated. PMID- 10691924 TI - Trypanosoma cruzi-induced immunosuppression: B cells undergo spontaneous apoptosis and lipopolysaccharide (LPS) arrests their proliferation during acute infection. AB - Acute infection with Trypanosoma cruzi is characterized by multiple manifestations of immunosuppression of both cellular and humoral responses. B cells isolated at the acute stage of infection have shown marked impairment in their response to polyclonal activators in vitro. The present work aims at studying the B cell compartment in the context of acute T. cruzi infection to provide evidence for B cell activation, spontaneous apoptosis and arrest of the cell cycle upon mitogenic stimulation as a mechanism underlying B cell hyporesponse. We found that B cells from acutely infected mice, which fail to respond to the mitogen LPS, showed spontaneous proliferation and production of IgM, indicating a high level of B cell activation. Furthermore, these activated B cells also exhibited an increase in Fas expression and apoptosis in cultures without an exogenous stimulus. On the other hand, B cells from early acute and chronic infected mice did not present activation or apoptosis, and were able to respond properly to the mitogen. Upon in vitro stimulation with LPS, B cells from hyporesponder mice failed to progress through the cell cycle (G0/G1 arrest), nor did they increase the levels of apoptosis. These results indicate that B cell apoptosis and cell cycle arrest could be the mechanisms that control intense B cell expansion, but at the same time could be delaying the emergence of a specific immune response against the parasite. PMID- 10691925 TI - Cross-reactivity of anti-galactocerebroside autoantibodies with a Trypanosoma brucei proteolipidic epitope. AB - Pathogenic mechanisms of the demyelinating encephalopathy featuring the nervous phase of human African trypanosomiasis (HAT) are largely unknown. They might include autoimmune disorders. A variety of autoantibodies is detected during the disease and we have previously evidenced anti-galactocerebroside (GalC) antibodies in the serum and cerebrospinal fluid (CSF) from patients in the nervous stage (stage II) of HAT. We now show that anti-GalC antibodies recognize an antigen located on the parasite membrane and common to different strains of trypanosomes. By using affinity chromatography with a rabbit anti-GalC antiserum, a 52-kD proteolipid was isolated from the membrane of Trypanosoma brucei (T. b.) brucei AnTat 1.9, AnTat 1. 1E, and T. b. rhodesiense Etat 1.2/R and Etat 1.2/S. Antibodies directed against this antigen were found in the CSF from patients with nervous stage HAT. These CSF also contained anti-GalC antibodies and adsorption with the proteolipid decreased anti-GalC reactivity. Immunization of mice with this antigen induced the production of antibodies which cross-reacted with GalC but no protection from experimental infection with T. b. brucei. These data support the hypothesis that anti-GalC antibodies detected in the CSF from HAT patients might be induced by molecular mimicry with a parasite antigen. PMID- 10691926 TI - The development of post-kala-azar dermal leishmaniasis (PKDL) is associated with acquisition of Leishmania reactivity by peripheral blood mononuclear cells (PBMC). AB - PKDL develops in about 50% of Sudanese patients treated for visceral leishmaniasis (kala-azar). Patients with kala-azar were entered into this study and followed for a period of up to 2 years. During follow up 12 patients developed PKDL and eight did not. Proliferative responses and cytokine production to Leishmania donovani and control antigens were measured in vitro using PBMC isolated at the time of diagnosis of kala-azar, after treatment of visceral leishmaniasis, during follow up, and at the time of diagnosis of PKDL. Proliferative responses and interferon-gamma (IFN-gamma) production were low at diagnosis and increased after treatment of kala-azar in both patients who developed (group 1) and those who did not develop PKDL later (group 2). In group 1, development of PKDL was always associated by an increased PBMC response to Leishmania antigen in proliferation and IFN-gamma production assays. There were no differences in Leishmania antigen-induced production of IL-4, IL-5 and IL-10 between or within the two groups. We have previously shown that Leishmania parasites spread to the skin during visceral leishmaniasis and proposed that PKDL was the result of an immunological attack on parasites, which have survived in the skin despite the drug treatment. The finding that PKDL develops after treatment of kala-azar as Leishmania-reactive T cells start to circulate in peripheral blood in sufficient numbers to be detected in in vitro assays supports this hypothesis. PMID- 10691927 TI - Autoantibodies to DEK oncoprotein in systemic lupus erythematosus (SLE). AB - Autoantibodies against the transcriptional DEK protein have been considered characteristic of the pauciarticular onset subtype of juvenile rheumatoid arthritis (JRA) associated with iridocyclitis in young girls. In this study we investigated the presence of anti-DEK autoantibodies in the sera of 288 patients with SLE using a recombinant DEK protein as autoantigenic target. Thirty sera (10.4%) were positive against DEK protein by immunoblotting. Patients with anti DEK autoantibodies show a lower frequency of cutaneous manifestation, exhibit more frequently certain markers of a chronic inflammatory status like anaemia and positivity for C-reactive protein, as well as a higher frequency of anti-double stranded DNA autoantibodies. In contrast to JRA patients positive for anti-DEK autoantibodies, no association with erosive arthritis nor iridocyclitis were found in SLE. In conclusion, our results show that 10.4% of SLE patients from our area show antibodies against DEK protein, although this feature did not clearly establish a clinical subset of the disease. PMID- 10691928 TI - Human monoclonal anti-phospholipid antibodies selectively bind to membrane phospholipid and beta2-glycoprotein I (beta2-GPI) on apoptotic cells. AB - The ability of an anti-phospholipid (LJ1) and an anti-beta2-GPI (RSP-57) human MoAb to bind to apoptotic but not viable cells was demonstrated in this study. Both MoAbs were derived from patients with systemic lupus erythematosus and anti phospholipid antibody syndrome. The parallel analysis of the specificity and affinity of four anti-phospholipid human MoAbs suggests that the binding of LJ1 MoAb to apoptotic cells is a specific property of this MoAb. RSP-57 MoAb recognizes apoptotic cells through beta2-GPI which becomes available for binding after the interaction with negatively charged phospholipids. This observation provides evidence that the binding of human anti-phospholipid antibodies to apoptotic cells occurs in both a beta2-GPI-dependent and independent way and involves a restricted group of epitopes. The finding that LJ1 and RSP-57 MoAbs bind apoptotic cells underlines the property of these MoAbs to act as cell membrane markers of apoptosis. Major pathological implications derive from the observation that LJ1 and RSP-57 MoAbs recognize epitopes expressed on 'early' apoptotic cells. The interference with the in vivo clearance and processing of apoptotic cells is a potential pathogenic mechanism of these antibodies. PMID- 10691929 TI - In the rheumatoid pannus, anti-filaggrin autoantibodies are produced by local plasma cells and constitute a higher proportion of IgG than in synovial fluid and serum. AB - IgG anti-filaggrin autoantibodies (AFA) are the most specific serological markers of rheumatoid arthritis (RA). They include the so-called 'anti-keratin antibodies' (AKA) and anti-perinuclear factor (APF), and recognize human epidermal filaggrin and other (pro)filaggrin-related proteins of various epithelial tissues. In this study we demonstrate that AFA are produced in rheumatoid synovial joints. In 31 RA patients, AFA levels were assayed at equal IgG concentrations in paired synovial fluids (SF) and sera. AFA titre-like values determined by indirect immunofluorescence and immunoblotting and AFA concentrations determined by ELISA were non-significantly different in serum and SF, clearly indicating that AFA are not concentrated in SF. In contrast, we demonstrated that AFA are enriched in RA synovial membranes, since the ELISA determined AFA in low ionic-strength extracts of synovial tissue from four RA patients represented a 7.5-fold higher proportion of total IgG than in paired sera. When small synovial tissue explants from RA patients were cultured for a period of 5 weeks, the profile of IgG and AFA released in the culture supernatants was first consistent with passive diffusion of the tissue infiltrating IgG (including AFA) over the first day of culture, then with a de novo synthesis of IgG and AFA. Therefore, AFA-secreting plasma cells are present in the synovial tissue of RA patients and AFA can represent a significant proportion of the IgG secreted within the rheumatoid pannus. PMID- 10691930 TI - PERB11 (MIC): a polymorphic MHC gene is expressed in skin and single nucleotide polymorphisms are associated with psoriasis. AB - The susceptibility genes for psoriasis remain to be identified. At least one of these must be in the major histocompatibility complex (MHC) to explain associations with alleles at human leucocyte antigen (HLA)-A, -B, -C, -DR, -DQ and C4. In fact, most of these alleles are components of just two ancestral haplotypes (AHs) designated 13.1 and 57.1. Although relevant MHC gene(s) could be within a region of at least 4 Mb, most studies have favoured the area near HLA-B and -C. This region contains a large number of non-HLA genes, many of which are duplicated and polymorphic. Members of one such gene family, PERB11.1 and PERB11.2, are expressed in the skin and are encoded in the region between tumour necrosis factor and HLA-B. To investigate the relationship of PERB11.1 alleles to psoriasis, sequence based typing was performed on 97 patients classified according to age of onset and family history. The frequency of the PERB11.1*06 allele is 44% in type I psoriasis but only 7% in controls (Pc = 0.003 by Fisher's exact test, two-tailed). The major determinant of this association is a single nucleotide polymorphism (SNP) within intron 4. In normal and affected skin, expression of PERB11 is mainly in the basal layer of the epidermis including ducts and follicles. PERB11 is also present in the upper keratin layers but there is relative deficiency in the intermediate layers. These findings suggest a possible role for PERB11 and other MHC genes in the pathogenesis of psoriasis. PMID- 10691931 TI - Cytomegalovirus immune globulin intravenous (human) administration modulates immune response to alloantigens in sensitized renal transplant candidates. AB - One of the important parameters for prolonged waiting time for potential renal transplant recipients is the presence of preformed antibodies to human leucocyte antigen (HLA) antigens, which is often caused by previous transplants, pregnancy or transfusions. In vivo administration of specific and unselected polyclonal intravenous immunoglobulin (IVIGs) preparations have been shown to inhibit anti HLA alloantibodies in highly sensitized patients. We sought to determine whether Cytogam (Medimmune Inc., Gaithersburg, MD, USA), a hyperimmune anticytomegalovirus immunoglobulin would (1) effect either in vitro or in vivo alloreactivity, and (2) whether Cytogam therapy could reduce the titre of preformed anti-HLA antibodies in highly sensitized patients. Alloreactivity was assessed by mixed lymphocyte reaction (MLR) and cytotoxic T lymphocyte (CTL) assay. A complement dependent microlymphocytotoxicity assay was done to assess for panel reactive antibody (PRA) status and the presence of anti-idiotypic antibodies in the Cytogam preparation. The MLR was inhibited by Cytogam in vitro in a dose-dependent fashion ranging from 31-92%. Significant inhibition of the MLR responses was not observed in recipients who received Cytogam in vivo (50 mg/kg). This could be a result of adminstration of a low dose of IVIG. However, CTL activity against the alloantigens in all individuals assessed was significantly inhibited after in vivo administration of Cytogam. Three of five individuals experienced a decrease of 5-32% in the PRA status at 4 weeks post administration of Cytogam. Cytogam also blocked the anti-HLA antibody titres in a microlymphocytotoxicity assay, suggesting the presence of anti-idiotypic antibodies. Our study was based on a single prophylactic dose of Cytogam (50 mg/kg), however, higher dose administration could be feasible by removing more fluid at dialysis, but should be given intradialytically to avoid volume overload. Overall, our results suggest that Cytogam can modulate the in vivo and in vitro T cell responses against the alloantigens. PMID- 10691932 TI - Staphylococcal acid phosphatase binds to endothelial cells via charge interaction; a pathogenic role in Wegener's granulomatosis? AB - The majority of patients with Wegener's granulomatosis (WG) are chronic nasal carriers of Staphylococcus aureus. Chronic nasal carriage of S. aureus is associated with an increased risk of developing a relapse of the disease. The mechanism by which this occurs is still unknown. We hypothesized that a cationic protein of S. aureus, staphylococcal acid phosphatase (SAcP), acts as a planted antigen and initiates glomerulonephritis and vasculitis in patients with WG. In order to test the hypothesis that SAcP can act as a planted antigen in WG, we studied the ability of SAcP to bind to human umbilical vein endothelial cells (HUVEC) and human glomerular endothelial cells. We also studied whether this binding can be prevented by preincubation with an anionic protein, and whether binding of SAcP activates endothelial cells. We also evaluated whether antibodies in sera of patients with WG are able to bind to endothelial cell-bound SAcP. The results show that SAcP can act as a planted antigen by binding to both types of endothelial cells in a concentration-dependent manner. Binding of concentrations as low as 4 microg/ml can be detected on HUVEC within 5 min of incubation. Binding of SAcP to endothelial cells was charge-dependent but did not activate endothelial cells. Finally, endothelial cell-bound SAcP was recognized by sera of patients with WG. The data suggest a possible pathogenic role for SAcP by acting as a planted antigen thereby initiating glomerulonephritis and vasculitis in patients with WG. PMID- 10691933 TI - Neutrophil FcgammaRIIIb allelic polymorphism in anti-neutrophil cytoplasmic antibody (ANCA)-positive systemic vasculitis. AB - Neutrophils constitutively express FcgammaRIIa and FcgammaRIIIb receptors. Both receptors exhibit allelic variants which have different quantitative functional capacities: the biallelic FcgammaRIIa-R131 and -H131 alleles, and the neutrophil antigen (NA) NA1/NA2 alleles. ANCA activation of neutrophils requires ligation of FcgammaRIIa receptor, but recent data have shown that ANCA can also bind FcgammaRIIIb receptor. The aim of this study was to determine whether the FcgammaRIIIb polymorphism was a risk factor for the development of ANCA associated systemic vasculitis, or the associated nephritis. FcgammaRIIIb receptor genotyping was determined by allele-specific polymerase chain reaction. Genomic DNA was extracted from 101 Caucasian patients with ANCA+ vasculitis (of whom 84 had renal disease) and 100 ethnically matched controls. Of the patients with ANCA+ systemic vasculitis, 71 had ANCA with specificity for proteinase 3 and 30 with specificity for myeloperoxidase (MPO). Overall no significant difference in genotype distribution or allele frequencies was found between patients and controls, or between patients with renal disease and controls. However, there was a trend for an increase in homozygosity for the NA1 allele in patients with a vasculitis and this was significant in patients who had anti-MPO antibodies. The FcgammaRIIIb receptor polymorphism is not a major factor predisposing to the development of ANCA+ systemic vasculitis or the associated nephritis. The over representation of the FcgammaRIIIb homozygous NA1 allele in patients with anti MPO antibodies may have implications for disease susceptibility. PMID- 10691934 TI - History of dermabrasion. PMID- 10691935 TI - Laser skin resurfacing of the face with a combined CO2/Er:YAG laser. AB - BACKGROUND: A combined, dual-wavelength CO2/Er:YAG laser system having the ability to deliver both clean ablation of skin with the erbium wavelength and a simultaneous deeper penetrating subablative thermal pulse of CO2 was developed for full-face resurfacing. The CO2 component can be pulsed from 1 to 100 msec at a power of 1-10 W with the Er:YAG component pulsed at 350 microsec at 1.7 J/cm2 through either a computer pattern generator with 3 mm diameter spot size or through a noncollimated spot ranging from 0.2 to 8 mm in diameter. Our previous study using this laser on the neck using a 4-8 mm diameter spot with Er:YAG fluence at 1.7 J and the CO2 at 5 W with a 50 msec pulse at a frequency of 10 Hz showed a higher degree of overall patient satisfaction, as well as improvement in skin texture and skin color, compared to patients treated with an Er:YAG laser alone. OBJECTIVE: This study evaluated the CO2/Er:YAG laser treatment modality in facial resurfacing. METHODS: Ten patients were treated with four passes at 1.7 J with a 4 mm diameter spot and the CO2 at 5 W with a 50-msec pulse at a frequency of 10 Hz. Photoaging scores as well as thermal damage and new collagen formation were compared immediately before and after treatment as well as at 2 weeks and 3 months postoperatively. RESULTS: The average pretreatment periorbital score was 6.2 The average posttreatment periorbital scores were 4.2 (P =.0239) at 2 weeks postoperatively (32% improvement) and 3.8 (P =.0028) at 3 months postoperatively (38% improvement). The average pretreatment perioral score was 5.9. The average posttreatment perioral scores were 3.0 (P =.0001) at 2 weeks postoperatively (49% improvement) and 3.3 (P =.0009) at 3 months postoperatively (44% improvement). The average pretreatment cheek score was 4.7. The average posttreatment cheek scores were 2.7 (P =.0066) at 2 weeks postoperatively (43% improvement) and 3.8 (P =. 0152) at 3 months postoperatively (36% improvement). The average pretreatment forehead score was 4.7. The average posttreatment forehead scores were 3.8 (P =.0340) at 2 weeks postoperatively (33% improvement) and 3.6 (P =.0147) at 3 months postoperatively (37% improvement). The average depth of collagen measured in the dermis pretreatment was 29 microm. The average depth of collagen 3 months posttreatment was 54 microm. This is an average increase of 25 microm or an 86% increase in collagen (P =.006). The average thermal damage immediately after treatment was 20 microm. CONCLUSION: The CO2/Er:YAG laser utilized with four passes at the above-mentioned parameters results in a similar degree of improvement as other forms of laser resurfacing with high-energy, short pulsed CO2 lasers. PMID- 10691936 TI - Sclerotherapy for treatment of hemangiomas. AB - BACKGROUND: While sclerotherapy in chronic venous insufficiency and in hemorrhoids is well established, the use of sclerotherapy for hemangiomas of the skin is widely unknown. OBJECTIVE: The aim of the study was to analyze the clinical value of sclerotherapy with polidocanol in a larger population. METHODS: Over a period of 20 years (1975-1995) we performed sclerotherapy of hemangiomas and vascular malformations in a total of 157 patients. Their ages ranged from 3 months to 75 years. Among these were 87 (55.4%) children and adolescents (up to the age of 18). Sclerotherapy with polidocanol was carried out mostly at monstrous or rapidly growing cavernous hemangiomas mainly localized in the face. RESULTS: One to three injections was usually sufficient to obtain the sclerosis effect; aesthetically, long-term results were convincing. Severe complications were not observed. CONCLUSION: Sclerotherapy of hemangiomas is a relatively simple, effective, and inexpensive method that is a valuable and promising treatment. PMID- 10691937 TI - Evaluation of a long-pulse Q-switched Nd:YAG laser for hair removal. AB - BACKGROUND: Hirsutism and hypertrichosis are common problems for which a permanent solution has been elusive. Laser-assisted hair removal is a promising technique. However, the optimal wavelength, pulse duration, and fluence continue to require further investigation. OBJECTIVE: To determine if the long-pulse Q switched Nd:YAG laser is safe and effective in reducing facial and non-facial trunk hair. METHODS: Fifteen patients were treated with a 30 msec pulsed Q switched Nd:YAG laser at fluences between 125 and 150 J/cm2. The reduction of hair density was assessed at baseline and at 7, 30, and 90 days after treatment. Potential complications were also evaluated. RESULTS: The average hair reduction was 36% at 7 days, 52% at 30 days, and 59% at 90 days. No significant complications or adverse events were reported. CONCLUSION: The long-pulse Q switched Nd:YAG laser provides a safe and effective means of hair removal. PMID- 10691938 TI - Treatment of facial skin using combinations of CO2, Q-switched alexandrite, flashlamp-pumped pulsed dye, and Er:YAG lasers in the same treatment session. AB - BACKGROUND: Many patients who seek facial CO2 laser resurfacing for improvement of photodamage are also concerned with "dark circles" under their eyes (periorbital hyperpigmentation) and/or telangiectasia as well as various types of deep scars on their faces. CO2 laser resurfacing alone provides limited improvement for these problems. OBJECTIVE: The purpose of this study was to demonstrate the conjunctive therapeutic effects of the CO2, Q-switched alexandrite, Er:YAG, and/or flashlamp-pumped pulsed dye lasers on facial skin treatments. METHOD: Thirty patients who underwent CO2 laser resurfacing were treated with additional lasers specific for their cosmetic concerns. Twenty patients with facial telangiectasias were treated with the pulsed dye laser immediately prior to CO2 laser resurfacing. Eleven patients with periorbital hyperpigmentation were treated with the Q-switched alexandrite laser immediately following use of the pulsed CO2 laser. Eight patients having sharply defined acne scars were treated with the Er:YAG laser following use of the CO2 laser. All patients had peripheral feathering performed with the Er:YAG laser. Nine patients were treated with all four lasers. RESULTS: In addition to significant improvement of the wrinkle scores from the CO2 laser resurfacing, patients had 75 100% clearing of the periorbital hyperpigmentation. All patients with facial telangiectasia showed virtually 100% improvement. All deep wrinkles and sharply defined scars responded with combined CO2/Er:YAG laser better than with CO2 laser resurfacing alone. All feathering was more uniform, with a more subtle transition to nontreated skin. There were no complications that could be attributed to the simultaneous use of multiple lasers. CONCLUSIONS: For patients who present with multiple cosmetic complaints, combined treatment using appropriate lasers offers excellent therapeutic outcome. PMID- 10691939 TI - The use of conscious sedation for outpatient dermatologic surgical procedures. AB - BACKGROUND: Dermatologic surgery has undergone increasing levels of sophistication over the past few decades. Commensurate with this demand, an established anesthesia technique called conscious sedation has been employed. OBJECTIVES: Methods for performing office-based conscious sedation are described. Recommendations are made regarding prerequisites for conscious sedation in an office setting, patient selection, complications management, and postoperative discharge requirements. CONCLUSION: The goals of anesthesia are to provide for patient safety and comfort, to increase patient acceptance of the procedure, and to enhance the surgeon's efficiency and satisfaction. PMID- 10691940 TI - Pedunculated malignant melanoma. AB - BACKGROUND: The pedunculated melanoma is an unusual variant of nodular melanoma that presents a challenge in staging and management. OBJECTIVE: We discuss the clinical and histopathologic characteristics of a case of pedunculated melanoma and present a brief review of the literature. METHODS: Routine stain with hematoxylin and eosin was performed on tissue specimens. RESULTS: The pedunculated melanoma was excised. Sentinel lymph node dissection was performed and was negative for the presence of melanoma. CONCLUSIONS: Pedunculated melanoma is a rare type of melanoma. Conventional staging methods for melanoma may not be reliable in this type of tumor. Complete workup, possibly including sentinel lymph node dissection, should be performed in all patients with pedunculated melanomas. PMID- 10691941 TI - Alexandrite laser hair removal is safe for Fitzpatrick skin types IV-VI. AB - BACKGROUND: Various lasers have been developed for epilation of unwanted hair. Most studies, however, have been done in white patients with minimal reference to dark-skinned individuals. OBJECTIVE: To determine the safety profile of a long pulsed alexandrite laser for hair removal in patients with Fitzpatrick skin types IV-VI exclusively. METHODS: Prospective clinical evaluation conducted from June 1998 to April 1999 at a referral private clinic. Prelaser skin testing was performed starting at 16 J/cm2 and energy fluence selected according to response. Complications were recorded at each visit. RESULTS: One hundred and fifty patients are reported (18 men and 132 women) ranging in age from 15 to 50 years, for a total of 550 treatment sites. Complications occurred in only 2% of cases. CONCLUSION: The long-pulsed alexandrite laser is safe for hair removal in darker skin tones. Prelaser skin testing was not helpful in this study, as there was no relationship between skin reaction and the incidence of complications. PMID- 10691942 TI - Cutaneous hypersensitivity reaction to injectable hyaluronic acid gel. AB - BACKGROUND: Injectable hyaluronic acid gel is a non-animal biomaterial used for soft tissue augmentation. OBJECTIVE: The dermal implantation of this naturally occurring polysaccharide is reported to be well tolerated by patients, with a longer duration in tissue than bovine collagen without any major local or systemic side effects. We report a case of an acute hypersensitivity reaction in a woman after her third injection for improvement of melolabial fold wrinkles. METHODS: An adverse granulomatous-like response to the intradermal injection of a modified hyaluronic acid gel is described. RESULTS: The patient developed indurated and erythematous papulocystic nodules in the melolabial folds bilaterally at the sites of injection. CONCLUSION: Injectable hyaluronic acid gel can be associated with severe allergic reactions and patients should be warned of this possible treatment side effect. PMID- 10691943 TI - Applicability of the sentinel node technique to Merkel cell carcinoma. AB - BACKGROUND: Merkel cell carcinoma (MCC) resembles malignant melanoma in several ways. Both are cutaneous lesions of the same embryonic origin. Both have an unpredictable biologic behavior, early regional lymph node involvement, early distant metastases, and high recurrence rate. OBJECTIVE: To apply the sentinel node technique described for melanoma to MCC in light of the common biologic features of these two tumors. METHODS: Preoperative lymphoscintigraphy, intraoperative lymphatic mapping, and sentinel node biopsy and frozen section histology were performed to guide the surgical treatment of three patients with MCC. RESULTS: Application of this approach in patients with MCC is feasible, reproducible, and seems reliable. CONCLUSION: The use of the sentinel node technique for MCC will reduce the number of unnecessary lymphadenectomies, will enable identification of microscopic metastases to lymph nodes, and will improve the stratification and accrual of patients into adjuvant treatment protocols. It may even lead to a survival benefit. PMID- 10691944 TI - Electrosurgical suspension apparatus. AB - BACKGROUND: Dermatologic surgeons commonly employ electrosurgery. OBJECTIVE: We describe a novel, yet simple, electrosurgical suspension apparatus and variations that facilitate the performance of excision and repair, Mohs micrographic surgery, cosmetic surgery, and other forms of dermatologic surgery. METHODS: The described techniques result from more than a decade of use and refinements in electrosurgical suspension apparatuses. RESULTS: The use of an electrosurgical suspension device has eliminated electrosurgical needle stick injuries, facilitated surgery, and reduced the risk of surgical field contamination via the electrosurgical handpiece or wiring. CONCLUSION: The use of a suspended electrosurgical handpiece results in reduced surgical time, eliminates many of the inconveniences associated with the current use of electrosurgery, and facilitates the use of electrosurgery as a tool of the dermatologic surgeon. PMID- 10691945 TI - Wire scalpel for surgical correction of soft tissue contour defects by subcutaneous dissection. AB - BACKGROUND: Increasing demand exists for cosmetic correction of soft tissue contour defects. Treatments include simple tissue augmentation techniques or more complex surgeries with consequent relevant recuperation time for the patient. The search for new simple techniques to correct scars and age-related wrinkles and folds is therefore one of the main goals of cosmetic dermatologic surgery. OBJECTIVE: To improve the cosmetic outcome of patients suffering from soft tissue contour defects by the use of a novel surgical instrument and technique, subcutaneous dissection by wire scalpel. METHODS: Fifty-four patients were treated with the wire scalpel technique with no skin incisions to correct a total of 132 depressed cosmetic defects of the face. Forehead lines, glabellar, nasolabial and oral commissure folds, upper lip wrinkles, and acne scars were treated. A 2-month to 4-year follow-up allowed subjective and photographic evaluation of results. RESULTS: Good or satisfactory results were obtained in 79.7% and 16.6% of the cases, respectively. Minor complications did not change the overall positive outcome of the surgery. CONCLUSION: Subcutaneous dissection by wire scalpel is a simple, safe, and effective method to improve the contour appearance of patients affected with scars or age-related contour defects. PMID- 10691946 TI - Sporadic Bazex-Dupre-Christol-like syndrome: early onset basal cell carcinoma, hypohidrosis, hypotrichosis, and prominent milia. AB - BACKGROUND: We present the case of a 32-year-old woman with a large recurrent multifocal basal cell carcinoma on the scalp. Conspicuous accompanying symptoms were multiple periorbital milia, hypotrichosis of the body and the scalp, and hypohidrosis. The sparse hair of the scalp showed further abnormalities such as pili torti, as well as flattened, irregularly curly hairs. OBJECTIVE: In 1964, Bazex et al. described a syndrome characterized by congenital hypotrichosis, follicular atrophoderma, and basocellular neoplasms that included basal cell nevi and early onset basal cell carcinomas. The Bazex-Dupre-Christol syndrome is a rare X-linked dominant disease. A sporadic occurrence with the typical constellation of these symptoms has not yet been reported. The lack of a positive family history and no signs of follicular atrophoderma argues for a sporadic occurrence of a Bazex-Dupre-Christol-like syndrome. The case reported shares several features with the classic Bazex-Dupre-Christol syndrome. CONCLUSION: Our report documents the necessity to look for early development of basal cell carcinomas in patients who show signs of the epidermal malformations described. PMID- 10691948 TI - Follicular unit hair transplanting--end of the evolution or a good thing taken too far? PMID- 10691947 TI - Placement of intravenous cannulae prior to ambulatory phlebectomy. AB - BACKGROUND: Ambulatory phlebectomy (AP) is a popular outpatient procedure for the removal of varicose veins. One of the major obstacles of AP is the disappearance and shift of the veins to be removed when the patient is positioned horizontally. OBJECTIVE: We aimed to verify the usefulness of preoperative placement of intravenous cannulae. METHODS: Forty-three vein segments of four consecutive patients with varicose veins were treated by AP with preoperative placement of intravenous cannulae. The procedures including anesthesia, incision, and exteriorization of the veins were performed with intravenous cannula in situ. We compared the courses of cannula and preoperative conventional marking of the veins. The time spent with extracting a vein was also compared with that spent with conventional procedure. RESULTS: The preoperative cannulation provides excellent fixation of varicose veins to their original position, at least at the puncture sites. The disagreement of courses between cannula and preoperative conventional marking was noticed in 47.8% of the vein segments from the knees and thighs, and in 15% from the lower legs. The time spent for extracting a vein was reduced by more than half with our method. CONCLUSION: Intravenous cannulation prior to AP is a simple procedure, but significantly improves the technique of AP by more accurately guiding the site of anesthesia and incision. PMID- 10691949 TI - In support of follicular unit transplantation. PMID- 10691950 TI - Sharquie's metal ring in skin surgery. PMID- 10691951 TI - Is curettage useful prior to performing Mohs or excisional surgery? When and how do you use it? PMID- 10691952 TI - Commentary PMID- 10691953 TI - Lessons on dermoscopy. Diagnosis: dysplastic nevus (so-called Clark nevus). PMID- 10691954 TI - Regarding successful treatment of spider leg veins. PMID- 10691955 TI - Response PMID- 10691956 TI - Regarding invasion of the lacrimal system by basal cell carcinoma. PMID- 10691957 TI - Regarding the use of topical retinoid acid after dermasanding. PMID- 10691958 TI - Regarding TCA-based Blue Peel. PMID- 10691959 TI - Response PMID- 10691960 TI - The art but not the science... PMID- 10691961 TI - Laser skin resurfacing: perspectives at the millennium. PMID- 10691962 TI - Reactive oxygen species released from granulocytes stimulate 5-lipoxygenase activity in a B-lymphocytic cell line. AB - B-lymphocytes express 5-lipoxygenase (5-LO) protein but cellular leukotriene production is suppressed by selenium-dependent peroxidases. Thus it was of interest to check whether reactive oxygen species (ROS) which are released under inflammatory conditions can stimulate B-lymphocyte 5-LO and counteract peroxidase mediated suppression of cellular 5-LO activity. It was found that 5-LO in the Epstein-Barr virus-transformed B-lymphocytic cell line BL41-E95-A is activated by addition of hydrogen peroxide or xanthine/xanthine oxidase and after increasing the oxidative state of the cell by azodicarboxylic acid bis(dimethylamide). Generation of endogenous ROS from mitochondria by antimycin A also lead to a threefold upregulation of 5-LO activity in B-cells. There was almost no detectable endogenous superoxide formation in BL41-E95-A cells after stimulation with 4beta-phorbol 12-myristate 13-acetate. Co-incubation experiments with BL41 E95-A cells and granulocytes demonstrated that granulocyte-derived ROS can activate B-lymphocyte 5-LO. Addition of superoxide dismutase and/or catalase to the B-lymphocyte/granulocyte co-incubations and to B-lymphocyte homogenates revealed that the 5-LO activation is due to the superoxide-derived release of hydroperoxides or hydrogen peroxide from granulocytes. The data suggest that ROS formation plays an important role in the regulation of cellular 5-LO activity in B-lymphocytes. As leukotrienes affect B-cell functions like cell proliferation, activation and maturation, this finding provides a new link between the formation of ROS and the regulation of immune responses. PMID- 10691963 TI - Kinetic characterization of the substrate specificity and mechanism of mushroom tyrosinase. AB - This paper reports a quantitative study of the effect of ring substituents in the 1-position of the aromatic ring on the rate of monophenol hydroxylation and o diphenol oxidation catalyzed by tyrosinase. A possible correlation between the electron density of the carbon atom supporting the oxygen from the monophenolic hydroxyl group and the V Mmax values for each monophenol was found. In the case of o-diphenols the same effect was observed but the size of the side-chain became very important. NMR studies on the monophenols justified the sequence of the V Mmax values obtained. As regards the o-diphenols, on the other hand, only a fair correlation between NMR and V Dmax values was observed due to the effect of the molecular size of the ring substituent. From these data, it can be concluded that the redox step (k33) is not the rate-determining step of the reaction mechanism. Thus, the monophenols are converted into diphenols, but the order of specificities towards monophenols is different to that of o-diphenols. The rate limiting step of the monophenolase activity could be the nucleophilic attack (k51) of the oxygen atom of the hydroxyl group on the copper atoms of the active site of the enzyme. This step could also be similar to or have a lower rate of attack than the electrophilic attack (k52) of the oxygen atom of the active site of oxytyrosinase on the C-3 of the monophenolic ring. However, the rate-limiting step in the diphenolase activity of tyrosinase could be related to both the nucleophilic power of the oxygen atom belonging to the hydroxyl group at the carbon atom in the 3-position (k32) and to the size of the substituent side chain. On the basis of the results obtained, kinetic and structural models describing the monophenolase and diphenolase reaction mechanisms for tyrosinase are proposed. PMID- 10691964 TI - Stabilization of NAD-dependent formate dehydrogenase from Candida boidinii by site-directed mutagenesis of cysteine residues. AB - The gene of the NAD-dependent formate dehydrogenase (FDH) from the yeast Candida boidinii was cloned by PCR using genomic DNA as a template. Expression of the gene in Escherichia coli yielded functional FDH with about 20% of the soluble cell protein. To confirm the hypothesis of a thiol-coupled inactivation process, both cysteine residues in the primary structure of the enzyme have been exchanged by site-directed mutagenesis using a homology model based on the 3D structure of FDH from Pseudomonas sp. 101 and from related dehydrogenases. Compared to the wt enzyme, most of the mutants were significantly more stable towards oxidative stress in the presence of Cu(II) ions, whereas the temperature optima and kinetic constants of the enzymatic reaction are not significantly altered by the mutations. Determination of the Tm values revealed that the stability at temperatures above 50 degrees C is optimal for the native and the recombinant wt enzyme (Tm 57 degrees C), whereas the Tm values of the mutant enzymes vary in the range 44-52 degrees C. Best results in initial tests concerning the application of the enzyme for regeneration of NADH in biotransformation of trimethyl pyruvate to Ltert leucine were obtained with two mutants, FDHC23S and FDHC23S/C262A, which are significantly more stable than the wt enzyme. PMID- 10691965 TI - The role of the membrane-bound tumour antigen, melanotransferrin (p97), in iron uptake by the human malignant melanoma cell. AB - Melanotransferrin (MTf) is a membrane-bound transferrin (Tf) homologue with several characteristics in common with serum Tf. MTf is found at high levels in melanoma cells and previous studies have shown that MTf can bind Fe. In addition, Chinese hamster ovary cells transfected with MTf transport Fe from 59Fe-citrate at greater rates than control cells. However, the role of MTf in the Fe uptake process of human melanoma cells remains unknown. In the present study we have characterized the role of MTf in Fe uptake by SK-Mel-28 melanoma cells in order to understand its function. Initial studies examined whether modulation of intracellular Fe levels using the Fe chelator desferrioxamine (DFO) or the Fe donor ferric ammonium citrate (FAC) could change MTf mRNA levels. In contrast to transferrin receptor (TfR) mRNA that increased after exposure to DFO and decreased after incubation with FAC, there was no change in MTf mRNA levels. In addition, compared to control cells, there was no alteration of 125I-labelled anti-MTf mAb-binding in cells exposed to DFO or FAC, suggesting no change in the number of MTf sites. Further studies examined the ability of DFO and FAC to modulate Fe uptake from 59Fe-citrate which is bound by MTf. In contrast to the effect of DFO or FAC at increasing and decreasing Fe uptake from 59Fe-Tf, respectively, DFO had no influence on 59Fe-citrate uptake, whereas FAC markedly increased it. Collectively, these studies suggest that MTf is not regulated in a manner similar to the TfR in response to cellular Fe levels. MTf can be removed from the membrane by phosphatidylinositol-specific phospholipase C (PtdIns-PLC). Preincubation of melanoma cells with PtdIns-PLC reduced anti-MTf mAb binding to 3% of the control, while PtdIns-PLC only slightly reduced 59Fe uptake from 59Fe citrate. These results suggest that MTf played only a minor role in Fe uptake from 59Fe-citrate by these cells. The expression of MTf mRNA (poly A+) was also examined in 50 human tissues and found to be markedly different to Tf mRNA or TfR mRNA. Surprisingly, MTf mRNA expression was widespread in normal tissues, and was observed at its highest levels in the salivary gland. In contrast to expectations, MTf mRNA expression was generally greater in adult than fetal tissues. PMID- 10691966 TI - N-Linked glycans of proteins from mitral valves of normal pigs and pigs affected by endocardiosis. AB - Endocardiosis, a degenerative and dystrophic process affecting cardiac valves and described in many mammalian species, is characterized by the accumulation of glycosaminoglycans, in particular hyaluronic acid, in the extracellular matrix. The glycoprotein patterns of pig mitral valves in normal animals and animals affected by endocardiosis were investigated. A different N-linked glycosylation pattern of glycoproteins was detected in affected valves compared with normal ones. In either normal or pathological species, the detected N-linked glycans were of the complex type. However, in samples from affected valves, sialic acid showed a prevalence of the alpha2,6 linkage to the galactosyl residue, whereas in normal samples the most frequent linkage was of the alpha2,3 type. In normal valves, the majority of complex oligosaccharides presented two outer branches with different degrees of fucosylation and sialylation, whereas in pathological samples we noted an increased number of glycans having up to four outer branches. PMID- 10691967 TI - Immunological detection of alkaline-diaminobenzidine-negativeperoxisomes of the nematode Caenorhabditis elegans purification and unique pH optima of peroxisomal catalase. AB - We purified catalase-2 of the nematode Caenorhabditis elegans and identified peroxisomes in this organism. The peroxisomes of C. elegans were not detectable by cytochemical staining using 3, 3'-diaminobenzidine, a commonly used method depending on the peroxidase activity of peroxisomal catalase at pH 9 in which genuine peroxidases are inactive. The cDNA sequences of C. elegans predict two catalases very similar to each other throughout the molecule, except for the short C-terminal sequence; catalase-2 (500 residues long) carries a peroxisomal targeting signal 1-like sequence (Ser-His-Ile), whereas catalase-1 does not. The catalase purified to near homogeneity from the homogenate of C. elegans cells consisted of a subunit of 57 kDa and was specifically recognized by anti (catalase-2) serum but not by anti-(catalase-1) serum. Subcellular fractionation and indirect immunoelectron microscopy of the nematode detected catalase-2 inside vesicles judged to be peroxisomes using morphological criteria. The purified enzyme (220 kDa) was tetrameric, similar to many catalases from various sources, but exhibited unique pH optima for catalase (pH 6) and peroxidase (pH 4) activities; the latter value is unusually low and explains why the peroxidase activity was undetectable using the standard alkaline diaminobenzidine-staining method. These results indicate that catalase-2 is peroxisomal and verify that it can be used as a marker enzyme for C. elegans peroxisomes. PMID- 10691968 TI - A novel alpha-amylase gene is transiently upregulated during low temperature exposure in apple fruit. AB - An alpha-amylase gene product was isolated from apple fruit by reverse transcriptase PCR using redundant primers, followed by 5' and 3' RACE. The gene is a member of a small gene family. It encodes a putative 46.9 kDa protein that is most similar to an alpha-amylase gene from potato (GenBank accession M79328). In apple fruit this new gene was expressed at low levels, as detected by reverse transcriptase PCR, in a number of plant tissues and during fruit development. Highest levels of mRNA for this transcript were observed 3 to 9 days after placing apple fruit at 0.5 degrees C. Phylogenetic analysis of amino acid sequence places the potato and apple proteins as a distinct and separate new subgroup within the plant alpha-amylases, which appears to have diverged prior to the split between monocotyledonous and dicotyledonous plants. These two divergent alpha-amylases lack the standard signal peptide structures found in all other plant alpha-amylases, and have sequence differences within the B-domain and C domain. However, comparisons with structures of known starch hydrolases suggest that these differences are unlikely to affect the enzymatic alpha-1,4-amylase function of the protein. This is the first report of upregulation of a dicotyledonous alpha-amylase in response to low temperature, and confirms the presence of a new family of alpha-amylases in plants. PMID- 10691969 TI - High resolution x-ray analysis of two mutants of a curaremimetic snake toxin. AB - A previous mutational analysis of erabutoxin a (Ea), a curaremimetic toxin from sea snake venom, showed that the substitutions S8G and S8T caused, respectively, 176-fold and 780-fold affinity decreases for the nicotinic acetylcholine receptor (AchR). In view of the fact that the side-chain of Ser8 is buried in the wild type toxin, we wondered whether these affinity changes reflect a direct binding contribution of S8 to the receptor and/or conformational changes that could have occurred in Ea as a result of the introduced mutations. To approach this question, we solved X-ray structures of the two mutants S8G and S8T at high resolution (0.18 nm and 0.17 nm, with R factors of 18.0% and 17.9%, respectively). The data show that none of the mutations significantly modified the toxin structure. Even within the site where the toxin binds to the receptor the backbone conformation remained unchanged. Therefore, the low affinities of the mutants S8T and S8G cannot be explained by a large conformational change of the toxin structure. Although we cannot exclude the possibility that undetectable structural changes have occurred in the toxin mutants, our data support the view that, although buried between loop I and II, S8 is part of the functional epitope of the toxin. PMID- 10691970 TI - Expression and regulation of alkaline phosphatases in human breast cancer MCF-7 cells. AB - The effect of retinoic acid and dexamethasone on alkaline phosphatase (AP) expression was investigated in human breast cancer MCF-7 cells. Cellular AP activity was induced significantly by retinoic acid or dexamethasone in a time dependent and dose-dependent fashion. A marked synergistic induction of AP activity was observed when the cells were incubated with both agents simultaneously. Two AP isozymes, tissue-nonspecific (TNAP) and intestinal (IAP), were shown to be expressed in MCF-7 cells as confirmed by the differential rate of thermal inactivation of these isozymes and RT-PCR. Based on the two-isozyme thermal-inactivation model, the specific activities for TNAP and IAP in each sample were analyzed. TNAP activity was induced only by retinoic acid and IAP activity was induced only by dexamethasone. Whereas dexamethasone conferred no significant effect on TNAP activity, retinoic acid was shown to inhibit IAP activity by approximately 50%. Interestingly, TNAP was found to be the only isozyme activity superinduced when the cells were costimulated with retinoic acid and dexamethasone. Northern blot and RT-PCR analysis were then used to demonstrate that the steady-state TNAP mRNA level was also superinduced, which indicates that the superinduction is regulated at the transcriptional or post transcriptional levels. In the presence of the glucocorticoid receptor antagonist RU486, the dexamethasone-mediated induction of IAP activity was blocked completely as expected. However, the ability of RU486 to antagonize the action of glucocorticoid was greatly compromised in dexamethasone-mediated superinduction of TNAP activity. Furthermore, in the presence of retinoic acid, RU486 behaved as an agonist, and conferred superinduction of TNAP gene expression in the same way as dexamethasone. Taken together, these observations suggest that the induction of IAP activity by dexamethasone and the superinduction of TNAP by dexamethasone were mediated through distinct regulatory pathways. In addition, retinoic acid plays an essential role in the superinduction of TNAP gene expression by enabling dexamethasone to exert its agonist activity, which otherwise has no effect. PMID- 10691971 TI - Ligand binding inhibitors of A1 adenosine receptor from Rana rugosa are phospholipase A2s. AB - Inhibitors of the A1 adenosine receptor were isolated from the skin extract of Korean frog, Rana rugosa. The frog-skin extract was prepared by an electrical shock and fractionated with C4 followed by C18 reverse-phase HPLC. Two A1 receptor inhibitors were isolated using a filter binding assay and the molecular masses of the proteins were estimated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry to be 15 347 and 15 404 Da, respectively. The inhibitory activity was also measured against other membrane receptors, such as the A2 adenosine receptor, muscarinic acetylcholine receptor and capsaicin receptor. Ligand binding to the A2 and muscarinic receptors was also severely inhibited by these proteins. However, they did not inhibit the functional activation of the capsaicin receptor by its ligand, capsaicin, suggesting that inhibition of ligand-receptor binding occurs specifically. Their N-terminal sequences were determined by Edman degradation. Surprisingly, they showed sequence similarity to the secretory protein, phospholipase A2 from various organisms. The phospholipase A2 activity of both proteins was tested using Dole's assay technique. Both proteins showed phospholipase A2 activity, and therefore, they were designated as PLA2-R1 and PLA2-R2, respectively. In addition, their ligand-binding inhibitory activity depended on their phospholipase A2 activity. This is the first finding that the frog secretes a phospholipase A2 similar to that of snake venoms, which posess inhibitory activity against the adenosine A1, adenosine A2 and muscarinic receptors. PMID- 10691972 TI - Characterization of the 5'-flanking region of the human multidrug resistance protein 2 (MRP2) gene and its regulation in comparison withthe multidrug resistance protein 3 (MRP3) gene. AB - The multidrug resistance proteins MRP2 (symbol ABCC2) and MRP3 (symbol ABCC3) are conjugate export pumps expressed in hepatocytes. MRP2 is localized exclusively to the apical membrane and MRP3 to the basolateral membrane. MRP2 mRNA is expressed at a high level under normal conditions, whereas MRP3 mRNA expression is low and increases only when secretion across the apical membrane by MRP2 is impaired. We studied some of the regulatory properties of the two human genes using transient transfection assays with promoter-luciferase constructs in HepG2 cells and cloned fragments of 1229 nucleotides and 1287 nucleotides of the MRP2 and MRP3 5' flanking regions, respectively. The sequence between nucleotides -517 and -197 was decisive for basal MRP2 expression. Basal promoter activity of MRP3 was only 4% of that measured for MRP2. At submicromolar concentrations, the histone deacetylase inhibitor trichostatin A reduced the MRP2 reporter gene activity and expression of the protein. Disruption of microtubules with nocodazole decreased gene and protein expression of MRP2 and increased MRP3 reporter gene activity. The genotoxic 2-acetylaminofluorene decreased the activity of the human MRP2 reporter gene construct, but increased MRP3 gene activity and enhanced the amounts of mRNA and protein of MRP2 and MRP3. Thus, regulation of the expression of these ATP-dependent conjugate export pumps is not co-ordinate, but in part inverse. The inverse regulation of the two MRP isoforms is consistent with their distinct localization, their different mRNA expression under normal and pathophysiological conditions, and their different directions of substrate transport in polarized cells. PMID- 10691973 TI - Purification, cloning and sequence analyses for pro-metalloprotease-disintegrin variants from Deinagkistrodon acutus venom and subclassification of the small venom metalloproteases. AB - Acidic and basic hemorrhagic metalloproteases were purified from the venom of Deinagkistrodon acutus (from Fujian Province, China) using gel filtration and anion exchange on FPLC and reversed-phase HPLC. Their hemorrhagic activities and N-terminal sequences were characterized. Extensive screening of the venom gland cDNA after PCR amplification resulted in the identification and sequencing of a total of seven cDNA clones encoding the multidomain precursors of six acidic and one alkaline low molecular mass metalloproteases. Two of the precursors contain a processable disintegrin domain. Disintegrins of 5 kDa were also purified from the venom. The partial amino-acid sequences and molecular masses determined by electrospray ionization mass spectrometry of the purified proteins specifically match those deduced from two of the cDNA sequences. Moreover, phylogenetic analyses based on 30 complete sequences of low molecular mass venom metalloproteases revealed that they may be classified into three functional subtypes: acidic hemorrhagins, basic and moderate hemorrhagins, and nonhemorrhagic enzymes. Subtype-specific amino-acid substitutions in the C terminal regions of the enzymes were highlighted to explore the structure activity relationships of the enzymes. PMID- 10691974 TI - Structural investigations of the CuA centre of nitrous oxide reductase from Pseudomonas stutzeri by site-directed mutagenesis and X-ray absorption spectroscopy. AB - Nitrous oxide reductase is the terminal component of a respiratory chain that utilizes N2O in lieu of oxygen. It is a homodimer carrying in each subunit the electron transfer site, CuA, and the substrate-reducing catalytic centre, CuZ. Spectroscopic data have provided robust evidence for CuA as a binuclear, mixed valence metal site. To provide further structural information on the CuA centre of N2O reductase, site directed mutagenesis and Cu K-edge X-ray absorption spectroscopic investigation have been undertaken. Candidate amino acids as ligands for the CuA centre of the enzyme from Pseudomonas stutzeri ATCC14405 were substituted by evolutionary conserved residues or amino acids similar to the wild type residues. The mutations identified the amino acids His583, Cys618, Cys622 and Met629 as ligands of Cu1, and Cys618, Cys622 and His626 as the minimal set of ligands for Cu2 of the CuA centre. Other amino acid substitutions indicated His494 as a likely ligand of CuZ, and an indirect role for Asp580, compatible with a docking function for the electron donor. Cu binding and spectroscopic properties of recombinant N2O reductase proteins point at intersubunit or interdomain interaction of CuA and CuZ. Cu K-edge X-ray absorption spectra have been recorded to investigate the local environment of the Cu centres in N2O reductase. Cu K-edge Extended X-ray Absorption Fine Structure (EXAFS) for binuclear Cu chemical systems show clear evidence for Cu backscattering at approximately 2.5 A. The Cu K-edge EXAFS of the CuA centre of N2O reductase is very similar to that of the CuA centre of cytochrome c oxidase and the optimum simulation of the experimental data involves backscattering from a histidine group with Cu-N of 1.92 A, two sulfur atoms at 2.24 A and a Cu atom at 2. 43 A, and allows for the presence of a further light atom (oxygen or nitrogen) at 2.05 A. The interpretation of the CuA EXAFS is in line with ligands assigned by site directed mutagenesis. By a difference spectrum approach, using the Cu K-edge EXAFS of the holoenzyme and that of the CuA-only form, histidine was identified as a major contributor to the backscattering. A structural model for the CuA centre of N2O reductase has been generated on the basis of the atomic coordinates for the homologous domain of cytochrome c oxidase and incorporating our current results and previous spectroscopic data. PMID- 10691975 TI - Use of proteoliposomes to generate phage antibodies against native AMPA receptor. AB - To isolate antibodies against ionotropic glutamate receptors (GluRs), we prepared a phage antibody library from mice immunized with proteoliposomes containing purified alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), a selective GluRD receptor. Specific binders were selected by repeated rounds of affinity panning against immobilized GluRD liposomes. Using this approach, we obtained a panel of high-affinity antibody fragments that immunoprecipitated both recombinant and native GluRD receptors, but not GluR6, a kainate receptor subunit with a 40% sequence similarity. The antibody fragments showed subunit selectivity, some being strictly specific for GluRD, whereas others also recognized the GluRB and GluRC but not GluRA subunits. Further experiments indicated that the epitopes recognized were conformational in nature and reside in the N-terminal extracellular 400-residue X domain of GluRD. Our results suggest that proteoliposomes, in combination with phage display technology, provide an effective tool for the generation of high-affinity conformation sensitive monoclonal antibodies against predetermined membrane proteins. PMID- 10691976 TI - Structure and dynamics of lipid-associated states of apocytochrome c. AB - Apocytochrome c (apocyt c), which in aqueous solution is largely unstructured, acquires an alpha-helical conformation upon association with lipid membranes. The extent of alpha-helix induced in apocyt c is lipid-dependent and this folding process is driven by both electrostatic and hydrophobic lipid-protein interactions. The structural and dynamic properties of apocyt c in lipid membranes were investigated by attenuated total reflection Fourier transform infrared spectroscopy combined with amide H-D exchange kinetics. Apocyt c acquires a higher content of alpha-helical structure with negatively charged membranes than with zwitterionic ones. For all membranes studied here, the helices of these partially folded states of apocyt c have a preferential orientation perpendicular to the plane of the lipid membrane. The H-D exchange revealed that a small fraction of amide protons of apocyt c, possibly associated with a stable folded domain protected by the lipid, remained protected from exchange over 20 min. However, a large fraction of amide protons exchanged in less than 20 min, indicating that the helical states of apocyt c in lipid membranes are very dynamic. PMID- 10691977 TI - The gene encoding polyneuridine aldehyde esterase of monoterpenoid indole alkaloid biosynthesis in plants is an ortholog of the alpha/betahydrolase super family. AB - The biosynthesis of the anti-arrhythmic alkaloid ajmaline is catalysed by more than 10 specific enzymes. In this multistep process polyneuridine aldehyde esterase (PNAE) catalyses a central reaction by transforming polyneuridine aldehyde into epi-vellosimine, which is the immediate precursor for the synthesis of the ajmalane skeleton. PNAE was purified from cell suspension cultures of Rauvolfia serpentina. The N-terminal sequence and endoproteinase LysC fragments of the purified protein were used for primer design and for the amplification of specific PCR products leading to the isolation of PNAE-encoding cDNA from a R. serpentina library. The PNAE cDNA was fused with a C-terminal His-tag, expressed in Escherichia coli and purified to homogeneity using Ni-affinity chromatography. The pure enzyme shows extraordinary substrate specificity, completely different to other esterases. Sequence alignments indicate that PNAE is a new member of the alpha/beta hydrolase super family. PMID- 10691978 TI - Inactivation of calcineurin by hydrogen peroxide and phenylarsine oxide. Evidence for a dithiol-disulfide equilibrium and implications for redox regulation. AB - Calcineurin (CaN) is a Ca2+-and calmodulin (CaM)-dependent serine/threonine phosphatase containing a dinuclear Fe-Zn center in the active site. Recent studies have indicated that CaN is a possible candidate for redox regulation. The inactivation of bovine brain CaN and of the catalytic CaN A-subunit from Dictyostelium by the vicinal dithiol reagents phenylarsine oxide (PAO) and melarsen oxide (MEL) and by H2O2 was investigated. PAO and MEL inhibited CaN with an IC50 of 3-8 microM and the inactivation was reversed by 2, 3-dimercapto-1 propane sulfonic acid. The treatment of isolated CaN with hydrogen peroxide resulted in a concentration-dependent inactivation. Analysis of the free thiol content performed on the H2O2 inactivated enzyme demonstrated that only two or three of the 14 Cys residues in CaN are modified. The inactivation of CaN by H2O2 could be reversed with 1,4-dithiothreitol and with the dithiol oxidoreductase thioredoxin. We propose that a bridging of two closely spaced Cys residues in the catalytic CaN A-subunit by PAO/MEL or the oxidative formation of a disulfide bridge by H2O2 involving the same Cys residues causes the inactivation. Our data implicate a possible involvement of thioredoxin in the redox control of CaN activity under physiological conditions. The low temperature EPR spectrum of the native enzyme was consistent with a Fe3+-Zn2+ dinuclear centre. Upon H2O2 mediated inactivation of the enzyme no significant changes in the EPR spectrum were observed ruling out that Fe2+ is present in the active enzyme and that the dinuclear metal centre is the target for the oxidative inactivation of CaN. PMID- 10691979 TI - DNase I hypersensitive sites and transcriptional activation of the lamin A/C gene. AB - The lamin A/C gene encodes subtypes of nuclear lamins, which are involved in nuclear envelope formation, and was recently identified as the responsible gene for the autosomal dominant Emery-Dreifuss muscular dystrophy. Expression of the lamin A/C gene is developmentally regulated but little is known about the regulatory mechanism. Previous studies of lamin A/C expression suggested that the chromatin structure is important for the regulation of its expression. To elucidate the regulatory mechanism of the lamin A/C gene expression, we have analysed the functional region of the mouse lamin A/C promoter and the chromatin structure of the gene in terms of nucleosome structure and DNase I hypersensitivity. Our analyses revealed disruption of the nucleosome array at the promoter region and the presence of multiple DNase I hypersensitive sites (HSs) which were specifically associated with expression of the lamin A/C gene. Inclusion of a segment which contained the HSs in a lamin A/C promoter-luciferase reporter plasmid showed no effect on the transfected promoter activity in transient expression assays. On the other hand, substantial enhancement of the promoter activity was detected when the transfected DNA was stably integrated into the genome, suggesting the importance of the HSs in the regulation of lamin A/C expression. PMID- 10691980 TI - Molecular mode of interaction of plant amine oxidase with the mechanism-based inhibitor 2-butyne-1,4-diamine. AB - 2-Butyne-1,4-diamine (DABI) is a mechanism-based inhibitor of copper-containing plant amine oxidases; the number of turnovers that leads to enzyme inactivation is approximately 20. The product of DABI oxidation is a very reactive aminoallene that reacts with an essential nucleophilic group at the enzyme active site, forming a covalently bound pyrrole and producing an inactive enzyme. The inactivated enzyme shows a new absorption maximum at 295 nm and gives coloured derivatives with p-dimethylaminobenzaldehyde and p-dimethylaminocinnamaldehyde that are spectrally similar to the products of pyrrole treated with the above reagents. Resonance Raman spectra of the p-dimethylaminobenzaldehyde adduct of pyrrole and the inactivated enzyme show very high degree of similarity, supporting the idea that the product of inactivation is indeed a bound pyrrole. The bound pyrrole is formed already in the anaerobic step of the reaction, while the topa semiquinone radical is not affected, as shown by the EPR and stopped flow absorption measurements. Peptides containing the DABI binding site were obtained by proteolysis of inactivated enzyme, isolated by HPLC and analysed by amino acid sequencing and MS. The crystal structure of the amine oxidase from pea has been determined; inhibition is caused mainly by the highly reactive DABI product, 4-amino-2-butynal, binding to a nucleophilic residue at the entrance to the substrate channel. As other DABI labelled peptides were also found and no free DABI product was detected by MS after complete inhibition of the enzyme, it is likely that the DABI product binds also to other solvent exposed nucleophilic residues on the enzyme surface. PMID- 10691981 TI - Biochemical and functional characterization of the Tn-specific lectin from Salvia sclarea seeds. AB - SSL, the lectin isolated from Salvia sclarea seeds, recognizes the Tn antigen (GalNAcalpha-O-Ser/Thr), a specific marker of many human carcinomas. Two dimensional electrophoresis, amino-acid and amino-sugar analysis, and MALDI-TOF MS showed that SSL is an acidic (pI 5.5), 60-61-kDa dimeric glycoprotein composed of apparently identical subunits linked by a single disulfide bond. The apparent molecular mass of SSL in solution determined by equilibrium sedimentation analytical ultracentrifugation was 59 +/- 9 kDa. This value did not change in the pH range 2.5-8.5, indicating that SSL does not associate into higher order structures. Tandem mass spectrometry and methylation analysis of N-glycans released from SSL by hydrazinolysis indicated that SSL possesses 2-3 glycosylation sites occupied with the typical plant glycans Manalpha1 6[(Manalpha1-3)(Xylbeta1-2)]Manbeta1-4 -GlcNAcbeta1-4(Fucalp ha1-3)GlcNAc and [(Manalpha1-3/6)(Xylbeta1-2)]Manbeta1-4-GlcNAcbeta1 -4(Fucalpha1-3)Glc NAc. The influence of adjacent Tn structures on the binding of two Tn-specific lectins (SSL and the isolectin B4 from Vicia villosa) and an anti-Tn monoclonal antibody (mAb 83D4) was evaluated using synthetic Tn glycopeptides. The binding of both lectins to the synthetic Tn glycopeptides was independent of the density of Tn structures. On the other hand, mAb 83D4 only reacted with glycopeptides displaying two or three consecutive Tn structures. PMID- 10691982 TI - His230 of serine hydroxymethyltransferase facilitates the proton abstraction step in catalysis. AB - The three-dimensional structures of rabbit and human liver cytosolic serine hydroxymethyltransferase revealed that H231 interacts with the O3' of pyridoxal 5'-phosphate and other residues at the active site such as S203, K257, H357 and R402 (numbering as per the human enzyme). This and the conserved nature of H231 in all serine hydroxymethyltransferases highlights its importance in catalysis and/or maintenance of oligomeric structure of the enzyme. In an attempt to decipher the role of H230 (H231 of the human enzyme) in the catalytic mechanism and/or maintenance of oligomeric structure of sheep liver serine hydroxymethyltransferase, the residue was mutated to arginine, phenylalanine, alanine, asparagine or tyrosine. Our results suggest that the nature of the amino acid substitution has a marked effect on the catalytic activity of the enzyme. H230R and H230F mutant proteins were completely inactive, dimeric and did not bind pyridoxal-5'-phosphate. On the other hand, mutation to alanine and asparagine retained the oligomeric structure and ability to bind pyridoxal-5' phosphate. These mutants had only 2-3% catalytic activity. The side reactions like transamination and 5,6,7, 8-tetrahydrofolate independent aldol cleavage were much more severely affected. They were able to form the external aldimine with glycine and serine but the quinonoid intermediate was not observed upon the addition of 5,6,7,8-tetrahydrofolate. Mutation to tyrosine did not affect the oligomeric structure and pyridoxal-5'-phosphate binding. The H230Y enzyme was 10% active and showed a correspondingly lower amount of quinonoid intermediate. The kcat / Km values for L-serine and Lallothreonine were 10-fold and 174-fold less for this mutant enzyme compared to the wild-type protein. These results suggest that H230 is involved in the step prior to the formation of the quinonoid intermediate, possibly in orienting the pyridine ring of the cofactor, in order to facilitate effective proton abstraction. PMID- 10691983 TI - Structure-function relationships of temporins, small antimicrobial peptides from amphibian skin. AB - Temporins, antimicrobial peptides of 10-13 residues, were isolated from secretions of Rana temporaria [Simmaco, M., Mignogna, G., Canofeni, S., Miele, R., Mangoni, M.L. & Barra, D. (1996) Eur. J. Biochem. 242, 788-792]. These molecules are specific to this amphibian species, which is also able to secrete on its skin other antimicrobial peptides similar to those found in different Rana species. The effect of temporins A, B and D (13 residues, net charge +2), and H (10 residues, net charge +1 and +2, respectively) against both artificial membranes of differing lipid composition and bacteria has been investigated in order to gain insight into their mechanisms of action. The results indicate that: the lytic activity of temporins is not greatly affected by the membrane composition; temporins A and B allow the leakage of large-size molecules from the bacterial cells; temporin H renders both the outer and inner membrane of bacteria permeable to hydrophobic substances of low molecular mass; and temporin D, although devoid of antibacterial activity, has a cytotoxic effect on erythrocytes. The results allow important conclusions to be drawn about the minimal structural requirements for lytic efficiency and specificity of temporins. PMID- 10691984 TI - Characterization of a novel type VII beta-turn conformation for a bio-active tetrapeptide rigin A synergy between theoretical and experimental results. AB - The conformational analysis of an immunomodulating tetrapeptide rigin (H-Gly-Gln Pro-Arg-OH), shown to possess diverse immunological activity, has been investigated both theoretically and experimentally for its conformational preferences. Unrestrained molecular dynamics simulation studies in implicit dimethylsulfoxide provide strong support for the existence of a significant population of ordered reverse turn structures for the major trans isomer. Of the three different energy minimized families, generated from computer molecular modelling, only one could be complemented by most of the 1D and 2D 1H NMR parameters obtained in dimethylsulfoxide-d6. A variable temperature NMR experiment in dimethylsulfoxide-d6 revealed that the preferred conformation is not stabilized by an intramolecular hydrogen bonding interaction. An analysis of the 2D ROESY experiment provides evidence in favour of an uncommonly observed, rather ill-defined type VII beta-turn structure. A survey of the observed specific inter-and intra-residue NOE connectivities and their comparison with one of the predicted low-energy conformations, demonstrates synergy between the theoretical molecular modelling and experimentally determined NMR spectral data. The primary structure, rather than long-range interactions, appears to be critical in determining the folding behaviour of the bio-active rigin. The present structural attributes may be valuable in peptide drug design and development of the rigin analogs having more effective stimulating activity. PMID- 10691985 TI - Trypanosoma brucei contains a 2,3-bisphosphoglycerate independent phosphoglycerate mutase. AB - Assays of phosphoglycerate mutase (PGAM) activity in lysates of bloodstream form Trypanosoma brucei appeared not to require exogenous 2,3-bisphosphoglycerate, thus suggesting that this protist contains an enzyme belonging to the class of cofactor-independent PGAMs. A gene encoding a polypeptide with motifs characteristic for this class of enzymes was cloned. The predicted T. brucei PGAM polypeptide contains 549 amino acids, with Mr 60 557 and pI 5.5. Comparison with 15 cofactor-independent PGAM sequences available in databases showed that the amino-acid sequence of the trypanosome enzyme has 59-62% identity with plant PGAMs and 29-35% with eubacterial enzymes. A low 28% identity was observed with the only available invertebrate sequence. The trypanosome enzyme has been expressed in Escherichia coli, purified to homogeneity and subjected to preliminary kinetic analysis. Previous studies have shown that cofactor-dependent and -independent PGAMs are not homologous. It has been inferred that the cofactor independent PGAMs are in fact homologous to a family of metalloenzymes containing alkaline phosphatases and sulphatases. Prediction of the secondary structure of T. brucei PGAM and threading the sequence into the known crystal structure of E. coli alkaline phosphatase (AP) confirmed this homology, despite the very low sequence identity. Generally, a good match between predicted (PGAM) and actual (AP) secondary structure elements was observed. In contrast to trypanosomes, glycolysis in all vertebrates involves a cofactor-dependent PGAM. The presence of distinct nonhomologous PGAMs in the parasite and its human host offers great potential for the design of selective inhibitors which could form leads for new trypanocidal drugs. PMID- 10691986 TI - The transbilayer distribution of phospholipids in disc membranes is a dynamic equilibrium evidence for rapid flip and flop movement. AB - We studied the transbilayer redistribution of phospholipids in bovine rod outer segment membranes on thoroughly washed, Ficoll-floated osmotically intact disc vesicles; freshly prepared membranes separated from the disc stack by osmotic shock; and intact disc stacks with a permeabilized plasma membrane (A-discs, B discs C-discs, respectively). In all cases, spin-labelled phospholipid analogues (SL-PL) with choline, serine and ethanolamine head groups (PtdCho, PtdSer and PtdEtn, respectively) were taken up into the outer leaflet of the membranes by > 90% and within less than 30 s after SL-PL addition, as deduced from the disappearance of spin-label from the suspension medium and from the specific ESR spectrum of membrane-associated spin-label. Using BSA extraction, the amount of SL-PL in the outer leaflet of the bilayer was determined. It decreased with a mean half-time of < 5 min at 25 degrees C, indicating rapid redistribution of all spin-labelled phospholipids into the inner leaflet of the disc membranes. After 1 h, PtdCho and PtdEtn were distributed almost symmetrically, whereas PtdSer was 35 : 65% (in/out). Using subsequent incubation with BSA, the outward movement (flop) of the analogues was observed directly, demonstrating that inward and outward movements proceed in thermodynamic equilibrium. No effect of N-ethylmaleimide or ATP on the redistribution could be measured, which makes it unlikely that energy consuming translocase or flippase processes are involved in the redistribution in the dark. We reason that the solubilization zone around the photoreceptor rhodopsin may be the locus of rapid redistribution of the highly unsaturated disc phospholipid. PMID- 10691987 TI - Ribonucleases expressed by human pancreatic adenocarcinoma cell lines. AB - Human ribonucleases have been considered as a possible tumor marker for pancreatic cancer, and elevated serum levels of ribonuclease activity in patients with pancreatic cancer have been reported by many authors. The reason for this elevation is unknown. In this study, we demonstrate that human pancreatic adenocarcinoma cell lines synthesize and secrete different ribonucleases. We isolated and characterized human pancreatic, or secretory, ribonuclease (RNase 1) from the conditioned media of the human pancreatic adenocarcinoma cell lines Capan-1, MDAPanc-3, IBF-CP3 and Panc-1, and the ampullary adenocarcinoma cell line MDAAmp-7, which represent a wide range of differentiation stages. Only one of these cell lines, Panc-1, produces significant amounts of nonsecretory ribonuclease. We then established a purification procedure for both secretory and nonsecretory ribonucleases, consisting of concentration of the supernatant by tangential filtration, anion-exchange and cation-exchange liquid chromatography and C4 RP-HPLC. Ribonuclease activity fractions were monitored using both the spectrophotometric and negative-staining zymogram techniques. The results of N terminal sequence analysis, kinetic analysis and endoglycosidase digestion studies indicate that the main ribonuclease secreted by all the cell lines is the secretory-type ribonuclease and that it is composed of several differently N glycosylated forms. Northern blot analyses confirm that some of the cell lines express secretory ribonuclease mRNA. The mRNA levels produced by Panc-1 and MDAPanc-28 are too low to be detected. Similar levels of expression of nonsecretory ribonuclease are found by Northern blot analysis in all the cell lines except Panc-1, which expresses higher levels. Here, we describe, for the first time, that several human pancreatic cancer cell lines with different degrees of differentiation express and secrete ribonucleases. This fact indicates that one origin of the elevated serum RNase levels in patients with pancreatic cancer are tumor cells. Analysis of the oligosaccharide moiety of the RNase 1 secreted by Capan-1 shows that it is highly glycosylated and its N-glycan chains are significantly different from that of the RNase 1 produced by normal pancreas. These results renew the possibility of using human serum RNase 1 determination as a tumor marker. PMID- 10691988 TI - Assessment of amino-acid substitutions at tryptophan 16 in alpha-galactosidase. AB - The tryptophan residue at position 16 of coffee bean alpha-galactosidase has previously been shown to be essential for enzyme activity. The potential role of this residue in the catalytic mechanism has been further studied by using site directed mutagenesis to substitute every other amino acid for tryptophan at that site. Mutant enzymes were expressed in Pichia pastoris, a methylotrophic yeast strain, and their kinetic parameters were calculated. Only amino acids containing aromatic rings (phenylalanine and tyrosine) were able to support a significant amount of enzyme activity, but the kinetics and pH profiles of these mutants differed from wild-type. Substitution of arginine, lysine, methionine, or cysteine at position 16 allowed a small amount of enzyme activity with the optimal pH shifted towards more acidic. All other residues abolished enzyme activity. Our data support the hypothesis that tryptophan 16 is affecting the pKa of a carboxyl group at the active site that participates in catalysis. We also describe an assay for continuously measuring enzyme kinetics using fluorogenic 4 methylumbelliferyl substrates. This is useful in screening enzymes from colonies and determining the enzyme kinetics when the enzyme concentration is not known. PMID- 10691989 TI - Contribution of C-tail residues of potato carboxypeptidase inhibitor to the binding to carboxypeptidase A A mutagenesis analysis. AB - The role of each residue of the potato carboxypeptidase inhibitor (PCI) C terminal tail, in the interaction with carboxypeptidase A (CPA), has been studied by the analysis of two main kinds of site-directed mutants: the point substitution of each C-terminal residue by glycine and the sequential deletions of the C-terminal residues. The mutant PCI-CPA interactions have been characterized by the measurement of their inhibition constant, Ki, in several cases, by their kinetic association and dissociation constants determined by presteady-state analysis, and by computational approaches. The role of Pro36 appears to be mainly the restriction of the mobility of the PCI C-tail. In addition, and unexpectedly, both Gly35 and Pro36 have been found to be important for folding of the protein core. Val38 has the greatest enthalpic contribution to the PCI-CPA interaction. Although Tyr37 has a minor contribution to the binding energy of the whole inhibitor, it has been found to be essential for the interaction with the enzyme following the cleavage of the C-terminal Gly39 by CPA. The energetic contribution of the PCI secondary binding site has been evaluated to be about half of the total free energy of dissociation of the PCI CPA complex. PMID- 10691990 TI - Nucleotide and amino-acid sequences of a new-type pectate lyase from an alkaliphilic strain of Bacillus. AB - A pectate lyase (pectate transeliminase; EC 4.2.2.2), designated Pel-15E, was purified to homogeneity from a culture broth of alkaliphilic Bacillus sp. strain KSM-P15. The purified enzyme had a molecular mass of approximately 33 kDa, as determined by SDS/PAGE, and a pI of approximately pH 9.2. Pel-15E exhibited optimum activity at pH 10.5 and 50-55 degrees C in glycine/NaOH buffer. Pel-15E had an absolute requirement for Ca2+ ions for manifestation of the enzymatic activity and trans-eliminated poly(galacturonic) acid, most likely by endo-type cleavage. A gene for the enzyme, which was cloned using the shotgun method and sequenced, contained a 960-bp ORF encoding 320 amino acids. The mature enzyme (286 amino acids, 32 085 Da) from the deduced amino-acid sequence showed quite low homology to known Pels from various microorganisms with 16.1-20.4% identity. Furthermore, we were not able to find any conserved regions in the sequence of Pel-15E when aligned with the sequences of other enzymes from the established Pel superfamily. However, Pel-15E had some regions that were homologous to PelA from Azospirillum irakense with 39.8% identity. Based on their amino-acid sequence homology, Pel-15E and PelA appear to belong to a new class of Pel family, although the enzymatic properties of both enzymes were quite different. PMID- 10691991 TI - Amino-acid sequence and glycan structures of cysteine proteases with proline specificity from ginger rhizome Zingiber officinale. AB - The ginger proteases (GP-I and GP-II), isolated from the ginger rhizome Zingiber officinale, have an unusual substrate specificity preference for cleaving peptides with a proline residue at the P2 position. The complete amino-acid sequence of GP-II, a glycoprotein containing 221 amino acids, and about 98% that of GP-I have been determined. Both proteases, which are 82% similar, have cysteine residues at positions 27 and histidines at position 161, corresponding to the essential cysteine-histidine diads found in the papain family of cysteine proteases, and six corresponding cysteine residues that form the three invariant disulfide linkages seen in this family of proteins. The sequence homology with other members (papain, bromelain, actinidin, protease omega, etc.) of this family is approximately 50%. GP-II has two predicted glycosylation sites at Asn99 and Asn156. Analyisis by electrospray and collision-induced dissociation MS showed that both sites were occupied by the glycans (Man)3(Xyl)1(Fuc)1(GlcNAc)2 and (Man)3(Xyl)1(Fuc)1(GlcNAc)3, in a ratio of approximately 7 : 1. Both glycans are xylose containing biantennary complex types that share the common core structural unit, Man1-->6(Man1-->3) (Xyl1-->2)Man1-->4GlcNAc1-->4(Fuc1-->3)GlcNAc for the major form, with an additional N-acetylglucosamine residue being linked, in the minor form, to one of the terminal mannose units of the core structure. PMID- 10691992 TI - In vitro study of proteolytic degradation of rat histidine decarboxylase. AB - Mammalian ornithine decarboxylase (ODC) is a very unstable protein which is degraded in an ATP-dependent manner by proteasome 26S, after making contact with the regulatory protein antizyme. PEST regions are sequences described as signals for protein degradation. The C-terminal PEST region of mammalian ODC is essential for its degradation by proteasome 26S. Mammalian histidine decarboxylase (HDC) is also a short-lived protein. The full primary sequence of mammalian HDC contains PEST-regions at both the N- and C-termini. Rat ODC and different truncated and full versions of rat HDC were expressed in vitro. In vitro degradation of rat ODC and rat 1-512 HDC were compared. Like ODC, rat 1-512 HDC is degraded mainly by an ATP-dependent mechanism. However, antizyme has no effect on the degradation of 1 512 HDC. The use of the inhibitors MG-132 and lactacystine significantly inhibited the degradation of 1-512 HDC, suggesting that a ubiquitin-dependent, proteasome 26S proteolytic pathway is involved. Results obtained with the different modifications of rat HDC containing all three PEST regions (full version, 1-656 HDC), only the N-terminal PEST region (1-512 HDC), or no PEST region (69-512 HDC), indicate that the N-terminal (1-69) fragment, but not the C terminal fragment, determines that the HDC protein is a proteasome substrate in vitro. PMID- 10691993 TI - Exhaled nitric oxide during incremental and constant workload exercise in chronic cardiac failure. AB - BACKGROUND: Nitric oxide (NO) is present in exhaled breath and produced by the pulmonary vascular endothelium as a potent vasodilator. Exercise is normally associated with pulmonary vasodilatation and a decrease in pulmonary vascular resistance to accommodate the increase in cardiac output. If production of NO is impaired in patients with chronic congestive cardiac failure (CCF), this might contribute to their exercise intolerance. PATIENTS AND METHODS: We quantified NO production (V NO) in 12 patients with chronic stable CCF and 12 controls, at rest and during incremental cardiopulmonary exercise on a treadmill, and at a later date during constant workload exercise. RESULTS: Patients had reduced V NO compared with controls during incremental exercise [381 (180) vs. 777 (275) nL min-1; mean (SD); P < 0.0001] but at constant workload V NO was similar between the two groups [353 (124) vs. 389 (189) nL min-1; P = 0.25]. Plasma levels of nitrate, the stable end-product of NO production, were significantly higher in patients [resting value 46.1 (21.6) vs. 23.0 (10.0) microM; P = 0.004] and were not influenced by exercise. CONCLUSION: Impaired NO-mediated pulmonary vasodilatation does not appear to contribute to exercise limitation in CCF. Alternatively, the lower NO production observed during maximal exercise in the patient group compared with controls may reflect a reduced incremental response of a system that is already abnormally activated in heart failure. PMID- 10691994 TI - Platelet factor 4 and beta-thromboglobulin in inflammatory bowel disease and giant cell arteritis. AB - BACKGROUND: As platelet factors are important in the inflammatory response, we examined the course of platelet factor 4 and beta-thromboglobulin in relation to disease activity in inflammatory bowel disease and in giant cell arteritis. PATIENTS AND METHODS: In a prospective study, the platelet count, platelet factor 4 and beta-thromboglobulin were measured in 20 patients with Crohn's disease, 18 with ulcerative colitis and 19 with giant cell arteritis, during active and inactive disease, as well as in 51 controls without inflammation. RESULTS: Platelet counts were significantly higher in active vs. inactive Crohn's disease, ulcerative colitis and giant cell arteritis. Levels of platelet factor 4 and beta thromboglobulin were significantly higher in active inflammatory bowel disease and giant cell arteritis, as well as in inactive inflammatory bowel disease and giant cell arteritis, than in the non-inflammatory controls. A positive correlation was found between the Crohn's disease activity index and the platelet count, platelet factor 4 and beta-thromboglobulin. Also, a positive correlation was found between the ulcerative colitis activity index and beta-thromboglobulin. However, even after 12 months of follow-up, in Crohn's disease and ulcerative colitis the mean levels of platelet factor 4 and beta-thromboglobulin were significantly higher than the levels of the controls. CONCLUSION: Platelet factors were correlated with inflammatory bowel disease activity. Levels of platelet factor 4 and beta-thromboglobulin, however, were markedly raised for a long time in clinically inactive inflammatory bowel disease, which might point to a pre-thrombotic state of disease. PMID- 10691995 TI - A metabolic syndrome of hypertension, hyperinsulinaemia and hypercholesterolaemia in the New Zealand obese mouse. AB - BACKGROUND: New Zealand obese (NZO) mice exhibit a polygenic obesity associated with hyperinsulinaemia and hyperglycaemia. Here we show that the strain presents additional features of a metabolic syndrome, i.e. elevated blood pressure, serum cholesterol and serum triglyceride levels. MATERIALS AND METHODS: A back-cross model of NZO mice with the lean Swiss Jackson Laboratory (SJL) strain was established in order to investigate further the correlation between hypertension, obesity, serum insulin and hyperglycaemia. RESULTS: Systolic blood pressure was significantly elevated at 6 weeks of age and appeared to parallel the weight gain of the animals. Serum insulin levels, presumably reflecting insulin resistance, and systolic blood pressure values were significantly correlated with the body mass index (r2 = 0.707 and 0.486, respectively) in the back-cross mice. In contrast, blood pressure was only weakly correlated with serum insulin (r2 = 0.288) in non-diabetic mice, and was independent of serum insulin levels in diabetic animals. CONCLUSION: The data are consistent with the concept that hypertension and insulin resistance are a characteristic consequence of the genetic constellation leading to obesity in the NZO strain, and that these traits reflect related mechanisms. It appears unlikely, however, that hypertension is a direct consequence of hyperinsulinaemia. PMID- 10691996 TI - Effect of tauroursodeoxycholic acid on bile acid-induced apoptosis in primary human hepatocytes. AB - BACKGROUND/AIMS: The accumulation of endogenous bile acids contributes to hepatocellular damage during cholestatic liver disease. To evaluate the potential role of apoptotic cell death due to increased concentrations of bile acids, primary human hepatocytes were treated with hydrophobic and hydrophilic bile acids. Because the Fas receptor-ligand system may mediate apoptosis in human liver cells, the effect of toxic bile acids on hepatocellular Fas receptor expression was evaluated. MATERIALS AND METHODS: Primary human hepatocytes were incubated with 50 and 100 microM glycochenodeoxycholic acid (GCDCA) and co incubated with equimolar concentrations of tauroursodeoxycholic acid (TUDCA). To evaluate cytolytic and apoptotic effects, morphological alterations, hepatocellular enzyme release, nuclear DNA fragmentation and hepatocellular Fas receptor expression were evaluated. RESULTS: Apoptotic cell death was significantly increased after exposure to 50 microM GCDCA. Bile acid-induced apoptosis was not accompanied by hepatocellular Fas receptor overexpression. Tauroursodeoxycholic acid reduced apoptosis, as indicated by a significant reduction of oligonucleosomal DNA cleavage. Fas receptor expression was not significantly affected by tauroursodeoxycholic acid. At higher concentrations, direct cytolytic cell destruction was observed. CONCLUSION: Primary human hepatocytes represent a suitable model to study bile acid-induced apoptotic cell death. In these hepatocytes, already low bile acid concentrations might induce apoptotic cell death, which is not triggered by hepatocellular Fas receptor overexpression. Apoptotic DNA fragmentation was significantly reduced by co incubation with tauroursodeoxycholic acid. The reduction of bile acid-induced apoptosis by ursodeoxycholic acid and its conjugates may contribute to the beneficial effects of these hydrophilic bile acids used for medical treatment of several cholestatic liver diseases. PMID- 10691997 TI - Speed of sound, bone mineral density and bone strength in oophorectomized rats. AB - BACKGROUND: The aim of this study was to determine the sensitivity of bone mineral density (BMD), ultrasounds (SOS) and resistance to torsion (T) to detect experimental osteopenia induced in rats 3 and 6 months after ooforectomy. MATERIALS AND METHODS: Seventy-four rats were used, divided into four groups, ooforectomized rats analysed 3 and 6 months after the operation and their respective control groups, in which BMD (Hologic QDR 1000 S/N 277), SOS (DBM Sonic 1200) and T (adapted test machine) were determined in the right femur. RESULTS: The results of the three techniques distinguished the ooforectomized groups from the controls, both 3 and 6 months after the ooforectomy, obtaining more significant differences with BMD (P = 0.0006, P = 0. 001, respectively) than SOS and T, where a significance of only P = 0.05 was obtained. In the correlation study among the three techniques, a significant correlation was observed between BMD and SOS (r = 0.39, P = 0.0008), as well as between BMD and T (r = 0.31, P = 0.03). However, significance was not observed between the SOS and T tests. CONCLUSION: In the study of sensitivity and specificity of the techniques used to detect the osteopenia caused by the ooforectomy, by means of calculation of the area under the receiver operation characteristic (ROC) curve, it was proven that although the three techniques distinguished between the two analysed populations, BMD presented an area under the ROC curve that was superior (0.87, 0.85) to that obtained with SOS (0.73, 0.67) and T (0.73, 0.68), both 3 and 6 months after the operation. PMID- 10691998 TI - Potential aetiological factors concerning the development of osteonecrosis of the femoral head. AB - BACKGROUND: The aetiology and pathogenesis of non-traumatic osteonecrosis (ON) of the femoral head have not been fully elucidated. The present study was conducted to evaluate the possible correlation of relevant haematological and biochemical factors with the development of ON. PATIENTS AND METHODS: Our investigation consisted of measurement of haematological indices and assessment of the biochemical and lipid profile of a study population of 68 patients with non traumatic ON of the femoral head and 36 healthy controls. The disease was considered idiopathic in 17 and secondary in 51 patients. RESULTS: There were no statistically significant differences in the parameters measured among the idiopathic ON, secondary ON and control groups, except for globulins alpha1, alpha2 and beta, which were significantly increased in both patient groups, and apolipoprotein B (Apo B), which was increased in patients with idiopathic disease compared with the control group. Both patient groups presented increased von Willebrand factor (VWF) and lipoprotein (a) [Lp(a)] levels and decreased protein C and S concentrations, but without statistical significance. However, both patient groups exhibited a greater proportion of abnormal values of any of these parameters, in 58.9% of the idiopathic and in 62.7% of the secondary ON patients, compared with 8.3% of the controls. CONCLUSION: Our study underlines the potential association of abnormal values of protein C, protein S, VWF and Lp(a) with ON. To our knowledge this is the first reported association of VWF with the disease. The majority of both idiopathic and secondary ON patients in our series exhibits a thrombotic potential that adds further support to the postulation that intravascular coagulation is a major pathogenetic mechanism leading to the disease. PMID- 10691999 TI - Expression of keratinocyte growth factor and its receptor in colorectal cancer. AB - BACKGROUND AND AIMS: The mesenchymal derived keratinocyte growth factor stimulates growth, differentiation and migration of intestinal epithelial cells. In the human gastrointestinal tract an overexpression of this growth factor has been reported in inflammatory bowel disease and pancreatic cancer. In the present study we investigated expression patterns of keratinocyte growth factor and receptor in normal and neoplastic colonic mucosa and in metastases. Furthermore, biological effects on normal intestinal and colorectal cancer cell lines were determined. MATERIALS AND METHODS: Expression patterns were analysed at the mRNA level by reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. Localization of ligand and receptor in normal intestinal mucosa and cancer tissue was investigated by immunohistochemistry. Mitogenic effects of keratinocyte growth factor were assayed by [3H]thymidine incorporation in normal (Intestine-407, IEC-6, IEC-18) and colorectal cancer cell lines (Colo320, LoVo, SW403, SW707). RESULTS: mRNA expression of keratinocyte growth factor and receptor was detected in the majority of normal and cancer samples without significant alterations. At the protein level keratinocyte growth factor expression did not differ between normal and malignant specimens, whereas protein expression of the receptor was increased up to twofold in well- to moderately differentiated colorectal cancers. DNA synthesis was significantly stimulated by keratinocyte growth factor in all three normal intestinal cell lines, whereas this growth factor did not significantly alter the [3H]thymidine incorporation in the colorectal cancer cell lines. CONCLUSION: Keratinocyte growth factor and its receptor were detected in the majority of samples from normal and neoplastic colonic mucosa, with an overexpression of the receptor seen in the more differentiated tumour samples. Keratinocyte growth factor is a strong mitogen for normal intestinal cells, whereas it is less effective in neoplastic cells. PMID- 10692000 TI - Lymphocyte responses following open and minimally invasive thoracic surgery. AB - BACKGROUND: Immunosuppression associated with surgery may predispose to increased tumour growth or recurrence. Lymphocytes are central components of the immune network, signalling specific and non-specific responses in tumour immunosurveillance. This study was therefore designed to compare the effects of minimally invasive and conventional approaches to major thoracic surgery on lymphocyte populations and oxidative activity. PATIENTS AND METHODS: The effects of conventional and minimally invasive video-assisted thoracic surgery (VATS) on the numbers and types of circulating lymphocytes and on lymphocyte oxidation were compared in a prospective randomized study of 41 patients undergoing lobectomy for peripheral bronchogenic carcinoma. Blood taken pre-operatively and on days 2 and 7 post-operatively was analysed for T (CD4, CD8), B (CD19) and natural killer (NK) (CD56, CD16) cell counts and for lymphocyte oxidative activity. Leucocyte numbers were compared with pre-surgical values and oxidative rate with healthy donor controls. RESULTS: Lymphocyte counts fell after surgery; VATS was associated with less effect on circulating T (CD4) cells at 2 days and on NK lymphocytes at 7 days post-surgery. Lymphocyte oxidation was less suppressed in the VATS group 2 days after surgery. In general, post-surgical changes in key cells of cellular immunity were smaller in the VATS group, and recovery to normal levels was more rapid. CONCLUSION: The degree of invasiveness of thoracic surgery may influence the extent of immunosuppression in patients undergoing pulmonary lobectomy for pulmonary neoplasm. PMID- 10692001 TI - Assessment of bilirubin toxicity to erythrocytes. Implication in neonatal jaundice management. AB - BACKGROUND: Neonatal hyperbilirubinaemia remains one of the most common clinical conditions requiring therapeutic intervention. Nevertheless, reliable indicators of bilirubin toxicity are still missing. This prompted us to investigate (a) the progression of cytotoxic events produced by increasing concentrations of bilirubin; (b) the relevance of the membrane lipid package on bilirubin binding to erythrocytes; and (c) the reliability of chloroform extraction compared with albumin extraction to evaluate erythrocyte-bound bilirubin and cytotoxicity. MATERIALS AND METHODS: Morphological alterations, free bilirubin, erythrocyte bound bilirubin (albumin- and chloroform-extractable), haemolysis and membrane released lipids, were determined in human erythrocytes at 4 degrees C or 37 degrees C, after 4 h incubation at pH 7.4, with increasing molar ratios of bilirubin to albumin (0.5-5). The reversibility of cytotoxicity by albumin washing was assessed by morphological analysis. RESULTS: Decreased free bilirubin, lower erythrocyte-bound bilirubin concentration by albumin extraction (superficial/non-aggregated bilirubin) and higher values by chloroform extraction (deep/aggregated bilirubin) were observed for 37 degrees C vs. 4 degrees C, at molar ratios > 1. Echinocytosis increased with bilirubin concentration and temperature and was not fully reversed by albumin washing. Haemolysis was already significant at a molar ratio of 1, and was enhanced by temperature at molar ratios 3 and 5 (P < 0.01). The loss of membrane lipids was remarkable at molar ratios > or = 0.5, both at 4 degrees C and 37 degrees C (P < 0.01), although correlation with bilirubin concentration was only significant at 37 degrees C (r = 0.971; P < 0.01). CONCLUSIONS: These results suggest that increased lipid fluidity and high bilirubin concentrations promote membrane bilirubin translocation and toxicity. They also show that albumin is not able to displace the bilirubin located deeply or aggregated within the membrane, which in turn is removed by chloroform. Accordingly, chloroform-extractable rather than albumin extractable bilirubin is a more accurate parameter to assess erythrocyte-bound bilirubin during severe hyperbilirubinaemia. PMID- 10692002 TI - Non-transferrin-bound iron is present in serum of hereditary haemochromatosis heterozygotes. AB - BACKGROUND: Hereditary haemochromatosis (HH) is a common autosomal recessive disease. Recently, HH heterozygosity has been identified as an independent risk factor for myocardial infarction and cardiovascular mortality. Iron may play an important role in atherogenesis by catalyzing peroxidation of low-density lipoprotein (LDL), an essential step in atherogenesis. In iron overload conditions, non-transferrin-bound iron (NTBI) is found in serum, which can catalyze lipid peroxidation. We investigated whether sera of HH heterozygotes contain more NTBI than sera of normal controls. METHODS: In 27 treated HH homozygotes, 22 HH heterozygotes and 17 healthy control subjects, conventional parameters of iron status (serum iron, transferrin saturation, serum ferritin) were measured. NTBI was detected using HPLC after addition of nitrilotriacetic acid and pretreatment with cobalt. RESULTS: The conventional parameters of iron status were similar in the HH heterozygous group and the control group. NTBI was significantly higher in homozygotes compared to heterozygotes (1.79 micromol L-1 vs. 0.51 micromol L-1, 95% CI of the difference = 0.6-1.95, P < 0.001), and controls (1.79 micromol L-1 vs. - 0.3 micromol L-1, 95% CI of the difference = 1.36-2.81, P < 0.001). The difference in NTBI between the heterozygous subjects and control subjects was also significant (0.51 micromol L-1 vs. - 0. 3 micromol L-1, 95% CI of the difference = 0.05-1.57, P < 0.05). CONCLUSION: Phlebotomy treated HH homozygotes maintain a high and potentially harmful serum NTBI. HH heterozygotes have a higher serum NTBI than normal controls. The reported increased risk of cardiovascular events in heterozygous haemochromatosis may be explained by NTBI-catalyzed LDL peroxidation. PMID- 10692003 TI - Decreased vitamin A levels in common variable immunodeficiency: vitamin A supplementation in vivo enhances immunoglobulin production and downregulates inflammatory responses. AB - BACKGROUND: Vitamin A has a broad range of immunological effects, and vitamin A deficiency is associated with recurrent infections. Common variable immunodeficiency (CVI) is a group of B-cell deficiency syndromes with impaired antibody production and recurrent bacterial infections as the major manifestations, but the immunological dysfunctions may also include T cells and macrophages. In the present study we examined the possible role of vitamin A deficiency in CVI. PATIENTS AND METHODS: We analysed plasma vitamin A levels in 20 CVI patients and 16 controls, and examined the relationships between vitamin A and clinical, immunological and metabolic parameters in CVI. In the six CVI patients with the lowest vitamin A levels we also studied the effect of vitamin A supplementation in vivo on several immunological functions in these patients. RESULTS: (i) The majority of CVI patients had decreased vitamin A levels compared with healthy controls, as found in both cross-sectional and longitudinal testing. (ii) Low vitamin A levels were associated with the occurrence of chronic bacterial infections and splenomegaly as well as high neopterin levels. Decreased levels of carrier protein and malabsorption were not observed. (iii) Vitamin A supplementation in patients with low vitamin A levels resulted in increased interleukin-10 (IL-10) and decreased tumour necrosis factor-alpha (TNFalpha) levels, as found in both plasma and monocyte supernatants, possibly favouring anti-inflammatory net effects. (iv) Vitamin A supplementation in vivo also enhanced anti-CD40-stimulated IgG production, serum IgA levels and phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cell (PBMC) proliferation. CONCLUSION: A considerable subgroup of CVI patients appears to be characterized by low vitamin A levels. Our findings support a possible role for vitamin A supplementation in CVI, perhaps resulting in enhanced immunoglobulin synthesis and downregulated inflammatory responses. PMID- 10692004 TI - Encapsulated fish oil enriched in alpha-tocopherol alters plasma phospholipid and mononuclear cell fatty acid compositions but not mononuclear cell functions. AB - BACKGROUND: Several studies have reported that dietary fish oil (FO) supplementation alters cytokine production and other functional activities of peripheral blood mononuclear cells (PBMC). However, few of these studies have been placebo controlled and few have related the functional changes to alterations in PBMC fatty acid composition PATIENTS AND METHODS: Healthy subjects supplemented their diets with 9 g day-1 of encapsulated placebo oil (3 : 1 mix of coconut and soybean oils), olive oil (OO), safflower oil (SO), evening primrose oil (EPO) or FO [providing 2.1 g eicosapentaenoic acid (EPA) plus 1.1 g docosahexaenoic acid (DHA) per day] for 12 weeks; the capsules also provided 205 mg alpha-tocopherol per day. Blood was sampled at 4-weekly intervals and plasma and PBMC prepared. Plasma phospholipid and PBMC fatty acid composition, plasma alpha-tocopherol and thiobarbituric acid-reactive substance concentrations, plasma total antioxidant capacity, the proportions of different PBMC subsets, the proportions of PBMC expressing the adhesion molecules CD2, CD11b and CD54, and PBMC functions (lymphocyte proliferation, natural killer cell activity, cytokine production) were measured. All measurements were repeated after a 'washout' period of 8 weeks. RESULTS: The placebo, OO and SO capsules had no effect on plasma phospholipid or PBMC fatty acid composition. The proportion of dihomo gamma-linolenic acid in plasma phospholipids was elevated in subjects taking EPO and was decreased in subjects taking FO. There was no appearance of gamma linolenic acid in the plasma phospholipids or PBMC in subjects taking EPO. There was a marked increase in the proportion of EPA in the plasma phospholipids (10 fold) and PBMC (four-fold) of subjects taking FO supplements; this increase was maximal after 4 weeks of supplementation. There was an increase in the proportion of DHA in plasma phospholipids and PBMC, and an approximately 20% decrease in the proportion of arachidonic acid in plasma phospholipids and PBMC, during FO supplementation. Plasma concentrations of alpha-tocopherol were significantly elevated during supplementation in all subjects and returned to baseline values after the washout period. There were no effects of supplementation with any of the capsules on total plasma antioxidant activity or plasma thiobarbituric acid reactive substances or on the proportion of different PBMC subsets, on the proportion of PBMC expressing adhesion molecules, on natural killer cell activity, on the proliferation of mitogen-stimulated whole blood cultures or PBMC, or on the ex vivo production of a range of cytokines by whole blood cultures or PBMC cultures stimulated by either concanavalin A or lipopolysaccharide. CONCLUSION: Supplementation of the diet with 3.2 g EPA plus DHA per day markedly alters plasma phospholipid and PBMC fatty acid compositions. The lack of effect of FO upon PBMC functions may relate to the level of alpha tocopherol included in the supplements. PMID- 10692005 TI - The evolutionary ecology of resistance to parasitoids by Drosophila. AB - Parasitoids are the most important natural enemies of many insect species. Larvae of many Drosophila species can defend themselves against attack by parasitoids through a cellular immune response called encapsulation. The paper reviews recent studies of the evolutionary biology and ecological genetics of resistance in Drosophila, concentrating on D. melanogaster. The physiological basis of encapsulation, and the genes known to interfere with resistance are briefly summarized. Evidence for within- and between-population genetic variation in resistance from isofemale line, artificial selection and classical genetic studies are reviewed. There is now firm evidence that resistance is costly to Drosophila, and the nature of this cost is discussed, and the possibility that it may involve a reduction in metabolic rate considered. Comparative data on encapsulation and metabolic rates across seven Drosophila species provides support for this hypothesis. Finally, the possible population and community ecological consequences of evolution in the levels of host resistance are examined. PMID- 10692006 TI - Spatial models for hybrid zones. AB - We introduce a spatially explicit model of natural hybrid zones that allows us to consider how patterns of allele frequencies and linkage disequilibria change over time. We examine the influence of hybrid zone origins on patterns of variation at two loci, a locus under selection in a two-patch environment, and a linked neutral locus. We consider several possible starting conditions that represent explicit realizations of two alternative scenarios for hybrid zone origins: primary intergradation and secondary contact. Our results indicate that in some circumstances, differences in hybrid zone origins will result in substantially different patterns of variation that may persist for thousands of generations. Our conclusions are generally similar to those previously derived from partial differential equations, but there are also some important differences. PMID- 10692007 TI - Reduction in fitness of flea beetles which are homozygous for an autosomal gene conferring resistance to defences in Barbarea vulgaris. AB - Major resistance genes are present in Danish flea beetle (Phyllotreta nemorum) populations, enabling the beetles to utilize a defended plant, Barbarea vulgaris ssp. arcuata, as a host plant, whereas this plant is unsuitable for beetles lacking the resistance genes. Two lines of beetles carrying a resistance gene have been established which are near-isogenic with a susceptible line. Larval survival of offspring from crosses between flea beetles carrying resistance genes and susceptible beetles, tested in bioassays on the defended B. vulgaris, and sex ratios of the survivors, were consistent with the presence of a dominant, autosomal resistance gene in each of the lines. An attempt to produce pure breeding lines for the autosomal genes revealed that beetles that are homozygous for the resistance gene suffer a high mortality. This result was repeatable for both lines, and when both resistant males and females were used in the crosses. The high mortality was also independent of the plant (defended B. vulgaris or suitable radish) on which the beetles were reared. The results suggest that the time of death of homozygous resistant beetles is variable. The spread and maintenance of resistance genes in flea beetle populations are discussed. PMID- 10692008 TI - Absence of single-locus complementary sex determination in the braconid wasps Asobara tabida and Alysia manducator. AB - In species with single-locus complementary sex determination (sl-CSD), sex is determined by multiple alleles at a single locus. In the haplodiploid Hymenoptera, sl-CSD results in females, if individuals are heterozygous at the sex locus, and in males, if individuals are hemizygous (haploid males) or homozygous (diploid males). Several hymenopteran species have been shown to have sl-CSD, but in several others sl-CSD is absent and the phylogenetic distribution remains unclear. In the family Braconidae, all four species tested so far were shown to possess sl-CSD. In this study, inbreeding experiments were used to test for the presence of sl-CSD in two species belonging to a subfamily of the Braconidae, Asobara tabida and Alysia manducator (Alysiinae). In both species inbreeding experiments showed no difference in brood size or sex ratio compared to the (outbred) control group. Furthermore, the sex ratios found in the inbreeding treatment differed significantly from the sex ratios expected under sl CSD. Therefore, we conclude that sl-CSD is absent in these species. This study is the first to show the lack of sl-CSD in species of the Braconidae family and that hymenopteran sex-determining mechanisms can vary, even within a family. PMID- 10692009 TI - Phylogenetic information in inter-SINE and inter-SSR fingerprints of the artiodactyla and evolution of the bov-tA SINE. AB - Various interspersed repeated sequences and elements (IRSs) can be utilized to generate PCR-based multilocus fingerprint profiles by amplifying the interelement segments, using primers matching the elements themselves. We assessed the utility of inter-IRS fingerprinting in phylogenetic comparisons among six artiodactyl species using several primers derived from two abundant genomic components: the Bov-tA short interspersed nuclear elements (SINEs) and simple sequence repeats or microsatellites (SSRs). Character- and distance-based analyses of the fingerprint data produced trees conforming to the established phylogenetic relationships of species. The strength of phylogenetic signal from different primers varied; combining data from different experiments resulted in robust trees. Within the Cervidae, the hierarchical relationship [(Odocoileus, Rangifer) Alces] was strongly supported. Both methods appear useful tools for systematic studies at time scales <30 Myr. To elucidate the material basis of inter-SINE fingerprints, we obtained the first sequences of the 'bovid' Bov-tA element also from two cervids (reindeer and white-tailed deer) and analysed their relationship to a number of paralogous bovid elements. The differences among sequences, both intra- and interspecific, were relatively high (mean 18.5%); the sequences showed no clear clustering with the species from which they had been isolated. Most individual elements probably date back to the cervid-bovid ancestor >25 Myr ago, which is in line with the observed fingerprint distributions. PMID- 10692010 TI - Chromosomal localization of cotransformed transgenes in the hexaploid cultivated oat Avena sativa L. using fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) was used to localize two transgenes (gus and bar), carried on plasmids pACT-1F and pUBA, respectively, on mitotic metaphase squashes of T1 plants of the cultivated hexaploid oat Avena sativa L. cotransformed by microprojectile bombardment of embryogenic callus. Among the eight progeny analysed by FISH in each of two lines, we detected plants null, hetero- and homozygous for the two genes in one line, and plants null and heterozygous for the two genes in the other line. Our results demonstrated that in the two independent transformation events, the gus and bar genes had inserted in the same position relative to each other. In each transformation event, the insertions occurred on D satellite (SAT) chromosomes bearing a C genome translocation. PMID- 10692011 TI - Do Wolbachia influence fecundity in Nasonia vitripennis? AB - This paper reports the influence of a vertically transmitted symbiont, Wolbachia, on host fitness in the parasitic wasp, Nasonia vitripennis. We measured fecundities of uninfected strains and strains infected with either two Wolbachia variants (wAv,wBv) or one (wAv or wBv). Preliminary tests suggested that double infected females produce more offspring on average than uninfected females. However, further studies failed to yield consistent fitness effects. To control for host genetic effects, the genotype of the double-infected and uninfected strain was 'replaced' with three different host genetic backgrounds by introgression. Contrary to previous results, we found no convincing evidence for positive fitness effects of Wolbachia in Nasonia vitripennis, once host genetic background was controlled for. It can be concluded that under the experimental design used here, the fecundity effects associated with Wolbachia in N. vitripennis are small or absent. PMID- 10692012 TI - MCE activities and malathion resistances in field populations of the australian sheep blowfly (Lucilia cuprina). AB - Malathion resistance has been shown to be the result of a single point mutation in the LcalphaE7 gene in four independently isolated chromosomes of Lucilia cuprina. The resultant amino acid substitution specifies high malathion carboxylesterase (MCE) activity. We have assayed MCE activities and resistance to malathion in three sets of field-derived samples, two sets of isogenic lines and five mass populations, and show that resistance to malathion in these samples is associated with high MCE activity in both sets of isogenic lines and four of the five mass populations. Additional mechanisms contributing to MCE activity or malathion resistance may be present in one of the mass populations. A second point mutation in LcalphaE7 is responsible for conferring diazinon resistance by encoding an increased organophosphate (OP) hydrolase activity. We also assayed diazinon resistances from the same three samples and show that diazinon and malathion resistances were in complete disequilibrium, with two exceptions. One exception involves the mass population with additional resistance mechanism(s) and the other involves three isogenic lines that are resistant to both insecticides. The molecular data for these lines suggest that they carry a duplication of the LcalphaE7 gene. PMID- 10692013 TI - Quantitative genetics of female choice in an ultrasonic pyralid moth, Achroia grisella: variation and evolvability of preference along multiple dimensions of the male advertisement signal. AB - The mating system of Achroia grisella (Lepidoptera: Pyralidae) is characterized by male ultrasonic advertisement signalling to which females orientate. Although males provide no direct, somatic benefits to their mates, females prefer males whose signal characters are more exaggerated than the population means. Previous studies showed that the signal characters influencing mate attraction are highly repeatable and heritable. We measured the phenotypic and additive genetic variances (heritability) of female preference in A. grisella, as this additive genetic variance is one of the genetic assumptions of indirect models of sexual selection. We determined the preference index of female A. grisella by repeated phonotaxis trials in which a choice of simulated male signals was presented. These playback experiments showed that female preference indices varied but were repeatable within individuals. Specifically, females differ in the relative importance of the several signal characters during mate assessment. A subsequent half-sib breeding design revealed an amount of additive genetic variance for the female preference index (h(s)2 = 0.212, SE = 0.1347, P = 0.0611; CVA = 0.1826). Our study highlights the importance of careful preparation of test signals and experimental design for quantifying individual variation in (female) preference along multiple signal dimensions. PMID- 10692014 TI - Virulence and molecular diversity of parthenogenetic root-knot nematodes, Meloidogyne spp. AB - Root-knot nematodes (RKN) are sedentary endoparasites causing severe damage to a wide variety of crops, including tomato. Among them, the parthenogenetic species Meloidogyne arenaria, M. incognita and M. javanica are of particular economic importance. The genetic diversity and relationships of 17 populations belonging to these three major species, either avirulent or virulent against the tomato Mi resistance gene, were investigated in order to determine whether (a)virulence of the nematodes could be related to their molecular fingerprints. Genomic polymorphisms between populations were assessed by using amplified fragment length polymorphism (AFLP) markers, and data were treated by means of a multiple correspondence analysis. A total of 1550 polymorphic amplified DNA fragments were identified and used to compute the relationships between the populations. As expected, the three RKN species were clearly distributed into distinct groups, but combination of data for virulence phenotypes and DNA markers showed that clustering of populations was not associated with their (a)virulence against the tomato Mi resistance gene. Such a lack of correlation indicates that most of the observed DNA polymorphism is independent of virulence, which is presumably under host selection. This result demonstrates that virulent populations do not share a common origin, and strongly suggests that they might have appeared late after the establishment of these clonal lineages, as the result of independent mutational events. PMID- 10692015 TI - Reproductive characteristics of the flower-breeding Drosophila hibisci Bock (Drosophilidae) in eastern Australia: within-population genetic determinants of ovariole number. AB - Genetic variation for ovariole number in the flower-breeding Drosophila hibisci was studied within populations obtained from three separate sites in the centre of the species distribution along the east coast of Australia. Heritability for ovariole number (adjusted for body size), derived from isofemale lines from each site, was estimated to be h2+/-SE = 0.564+/-0.160. The variance of ovariole number within sites (sigma2(within) = 2.039) was comparable to the variance between sites (sigma2(between) = 2.048) obtained from an earlier study of populations sampled over 14 degrees of latitude. Two isofemale lines (from within one site) that differed by an average of 4.6 ovarioles were used to generate F1, F2 and backcross generations. Analysis of mean ovariole number for these generations showed that only additive gene effects were important and that dominance, digenic epistasis and maternal effects were not significant. This within-population result contrasted with earlier results between populations that revealed additive and digenic epistasis for the same trait. High heritability within populations and the relatively large within-population variation for ovariole number suggest that substantial microhabitat variation is influencing this fitness-related trait. PMID- 10692016 TI - Are the same genes responsible for intra- and interspecific variability for sex comb tooth number in Drosophila? AB - The identification of genes contributing to speciation has the potential to provide important insights into the mechanisms of evolution. One of the most interesting unresolved puzzles is the relationship between intraspecific variability in morphological traits and their interspecific divergence. Intraspecific polymorphisms without major detrimental side-effects may serve as a substrate for selection response during speciation. The same quantitative trait loci (QTLs) may, then, account for the trait variability both within and between species. In contrast, the vast majority of intraspecific variants could be deleterious mutations that have not yet been selected out. In this case intraspecific variation would not ultimately generate interspecific trait differences. In previous work, QTLs responsible for morphological differences between animal species, including those for the secondary sexual trait sex comb tooth number, have been mapped with the resolution of chromosome segments. Here, we mapped QTLs for which alleles segregated between two laboratory lines of Drosophila melanogaster. The two QTLs identified mapped to the X chromosome and accounted for only 8% of the between-line differences, implying that a large number of small-effect genes modify sex combs. One intraspecific QTL mapped to the same interval as the QTL for interspecific differences between D. simulans and D. mauritiana. Whether or not these effects result from the same genes requires further examination. PMID- 10692017 TI - Differences in genetic structure between two Japanese beech (Fagus crenata Blume) stands. AB - To examine the effects of forest cutting on within-population genetic structure, the genetic structure and variability of two Japanese beech (Fagus crenata Blume) stands with contrasting histories in relation to cutting were investigated. Six hundred and sixty beech trees, covering two hectares in total, were mapped and genetically analysed using nine isozyme loci encoding eight enzyme systems. The proportion of polymorphic loci, the average number of alleles per locus, the effective number of alleles per locus, the expected heterozygosity and the observed heterozygosity were 78, 3.3, 1.31, 0.200 and 0.189, respectively, in a secondary stand (designated AK) cut during the 1920s. Corresponding figures were 78, 3.3, 1.33, 0.203 and 0.193, respectively, in a primary stand designated KU. The inbreeding coefficient and the grand mean of the number of alleles in common (NAC) were 0.055 and 1.684 in AK, and 0. 042 and 1.649 in KU, respectively. The genetic variability was slightly but significantly lower in AK. The genetic structure of the two stands was strikingly different. The proportions of positively significant Moran's I and SND values found in the shortest distance class were 0.86 and 0.38 for AK, and 0.14 and 0.29 for KU, respectively. Furthermore, significant linkage disequilibrium was observed in AK, but none at all in KU. To examine which, if any, differences in the genetic structure would be likely to influence succeeding generations, we simulated a self-thinning process. The simulation suggested that reduced genetic variability and linkage disequilibrium would have significant influence in the AK stand for several generations. PMID- 10692018 TI - The effect of ecological factors on the mating system of a South American shrub species (Helicteres brevispira). AB - Mating systems are influenced by several ecological factors, including plant density, number of flowers per plant, and pollinator movements. In this study, we investigated the simultaneous effects of these three factors on the mating system of a self-compatible Brazilian shrub species: Helicteres brevispira St. Hil. Outcrossing rate is directly correlated with plant density. Changes in the number of flowers per plant affect outcrossing rate through their effect on the density of flowers. Variation in foraging behaviour of hummingbird pollinators is a consequence of the interaction between plant density and number of flowers per plant. Territorialist pollinators are common in high density areas but visit few flowers on each plant, thereby promoting outcrossing. In areas of low plant density, trapliners and rare territorialists visit several flowers per plant, thus increasing selfing. Our results indicate that outcrossing rate is a dynamic parameter, with the extent of variation depending on a number of ecological factors. In successional species such as those in the genus Helicteres, demographic changes may be accompanied by alterations in mating system parameters, with concomitant effects on the genetic structure of populations. PMID- 10692019 TI - MtDNA variation in Apis cerana populations from the Philippines. AB - The cavity-nesting honeybee Apis cerana occurs in Asia, from Afghanistan to China and from Japan to southern Indonesia. Based on morphometric values, this species can be grouped into four subspecies: A. c. cerana, A. c. indica, A. c. japonica and A. c. himalaya. In order to analyse the geographical variability of A. c. indica from the Philippine Islands, 47 colonies from different locations in three of the larger islands (Mindanao, Luzon and Palawan) and four of the Visayan Islands (Panay, Negros, Cebu and Leyte) were studied. Genetic variation was estimated by restriction and sequence analysis of PCR-amplified fragments of the tRNAleu-COII region. We found four different haplotypes, Ce1, Ce2, Ce3 and Ce4, that discriminate among the bee populations from different islands. The Ce1 haplotype is present in Mindanao and Visayan Islands, Ce2 is restricted to Luzon, and both Ce3 and Ce4 are only present in Palawan. Phylogenetic analysis of the sequences shows a great intraspecific variability, is in accordance with the geological history of these islands and partially agrees with some previous morphological and molecular studies. PMID- 10692020 TI - Surgical pathology of renal epithelial neoplasms: recent advances and current status. PMID- 10692021 TI - Metaplastic carcinoma of the breast arising within complex sclerosing lesion: a report of five cases. AB - AIMS: This study presents a series of five cases in which metaplastic carcinoma, predominantly low-grade adenosquamous carcinoma, of the breast is seen arising within a background of a complex sclerosing lesion. This association has been recognized previously but has not been documented in detail. This study describes the characteristics of the components present in each case and discusses the existing literature. This observation adds further evidence to support an association between some types of invasive breast carcinoma and sclerosing lesions of the breast. METHODS AND RESULTS: Four of these cases were received as referral cases for opinion. The fifth was received as part of the routine surgical workload within our own institution. Two patients presented following mammographic screening and three symptomatically; their mean age was 62 years (range 49-68). The mean lesion size was 16 mm (range 7-24). All five lesions showed features of a complex sclerosing lesion/radial scar in the form of central sclerosis with elastosis and radiating benign entrapped tubules. One had associated benign papillary structures and two had focal benign squamous metaplasia. Four cases showed coexisting but distinct areas of low-grade adenosquamous carcinoma with glandular and squamous epithelial differentiation in a spindle cell background. One case had associated undifferentiated spindle cell carcinoma. Detailed immunophenotypic characteristics of two cases are presented. CONCLUSIONS: This series illustrates a postulated but previously unconfirmed association between an unusual form of metaplastic breast carcinoma (adenosquamous carcinoma) and complex sclerosing lesions. The mechanisms of induction of breast carcinoma are poorly understood but these observations further emphasize the potential for sclerosing lesion of the breast to be associated with, and possibly give rise to, invasive carcinoma of different types. The precise nature of the interaction between the pathological processes remains unclear. PMID- 10692022 TI - Leiomyosarcomas of the oral cavity: an unusual topographic subset easily mistaken for nonmesenchymal tumours. AB - AIMS: Oral leiomyosarcoma is rare and poorly documented. We aimed to characterize these lesions clinicopathologically in order to facilitate their distinction from other spindle cell neoplasms in the oral cavity. METHODS AND RESULTS: Ten cases of oral leiomyosarcoma were retrieved and studied histologically and immunohistochemically. Clinical data were obtained from referring pathologists and prior literature concerning 46 comparable cases was reviewed. Nine out of 10 cases occurred in adults; 50% arose in the jaws and four showed bone involvement. Histological appearances were similar to leiomyosarcomas elsewhere. In addition to myogenic markers, two cases were also keratin-positive. Four patients developed local recurrence or metastatic disease and three died of tumour (median follow-up 37 months). CONCLUSIONS: Leiomyosarcoma is under-recognized in the mouth, often being mistaken for a spindle-celled epithelial neoplasm. Aside from an unusual but infrequent tendency to spread to lymph nodes and a location specific differential diagnosis, its clinicopathological features are comparable to leiomyosarcomas at other locations. PMID- 10692023 TI - Paratesticular liposarcoma with smooth muscle differentiation mimicking angiomyolipoma. AB - AIMS: To discuss the differential diagnosis of a case of well-differentiated liposarcoma which had areas resembling angiomyolipoma-a feature which, to our knowledge, has not been reported previously. METHODS AND RESULTS: A tumour in the paratesticular region had apparently been present for 40 years, but had grown recently. A fat component containing lipoblasts was admixed with areas resembling angiomyolipoma, i.e. desmin positive, but HMB45-negative smooth muscle proliferation with atypia and thick-walled blood vessels devoid of elastin. CONCLUSION: The diagnosis of liposarcoma, rather than angiomyolipoma with adipose atypia, in this case is based on the fact that smooth muscle differentiation is documented in liposarcoma, lack of HMB45 staining and recent clonality studies which suggest that the fat in angiomyolipoma is not neoplastic. PMID- 10692024 TI - Multinucleated stromal cells of the anal mucosa: a common finding. AB - AIMS: To document the presence, morphology, immunophenotype and ultrastructure of multinucleated stromal cells within the anal mucosa and to discuss possible pathogenetic mechanisms for this occurrence. METHODS AND RESULTS: Multiple sections of normal anal mucosa from 30 abdominoperineal resection specimens were analysed by light microscopic, electron microscopic and immunohistochemical methods. Multinucleated stromal cells were found in 22 cases (73%). They contained two to five nuclei, arranged in a linear fashion or in a rosette or grape-like pattern. They stained positive for vimentin and negative for actin, desmin and oestrogen/progesterone receptors. Ultrastructural examination confirmed their fibroblastic lineage. Mast cells were frequently observed in the immediate vicinity of mono- and multinucleated cells. CONCLUSIONS: Multinucleated stromal cells are a common occurrence in the normal anal mucosa. They should not be misinterpreted as neoplastic cells. Mast cells may play a role in their morphogenesis. PMID- 10692025 TI - CD101 expression by Langerhans cell histiocytosis cells. AB - AIMS: Our objective was to study the expression of a recently identified cell surface molecule, CD101 and in Langerhans cell histiocytosis (LCH) patients as CD101 has been shown to be present on dendritic cells. We wanted to determine if CD101 expression could be helpful for the diagnosis of LCH in conjunction with other markers (CD1a, S100 protein), and could be predictive of the evolution and dissemination of the disease. METHODS AND RESULTS: The expression of CD101 was studied by immunohistochemical technique in 11 cases of Langerhans cell histiocytosis on frozen sections. The expression of CD101 was positive in nine cases, high in six cases and low in three cases. There was no expression in the other two cases. No correlation with the evolution, the localization or the dissemination of the disease could be evidenced. CONCLUSIONS: CD101 is a new phenotypic marker that might be useful in combination with other markers for the diagnosis of LCH. However, as the anti-CD101 antibody works only in frozen sections, its value is limited compared to anti-CD1a antibody. PMID- 10692026 TI - Variation in the histological pattern of nodal involvement by gamma/delta T-cell lymphoma. AB - AIMS: Gamma-delta (gammadelta) T-cell non-Hodgkin's lymphomas (NHLs) usually present with liver, spleen and marrow infiltration. Lymph node involvement by gammadelta T-cell NHL has been rarely documented so far; its histological pattern needs to be further defined. METHODS AND RESULTS: Two cases of nodal gammadelta T cell NHL are reported: case 1, a 44-year-old man, presented with cytomegalovirus retinitis and superficial lymphadenopathies. Histological analysis of an inguinal lymph node showed complete destruction by a diffuse pleomorphic lymphoid proliferation, which was positive for CD2, CD3, CD43, CD45, TIA-1 and granzyme B, and displayed a gammadelta phenotype (deltaTCR1+, Vdelta1+, Vdelta2-, Vdelta3-, betaF1-). Bone marrow was normal. Case 2, a male 24-year-old patient with a history of renal transplantation, presented with hepatosplenomegaly and supraclavicular lymph node enlargement. Lymph node architecture was globally preserved. Peripheral sinuses contained scattered nests of medium-sized irregular lymphoid cells. Bone-marrow was infiltrated. Phenotype showed positivity for CD2, CD3, CD45 and TIA1 and expression of gammadelta TCR (deltaTCR1+, deltaV1+, deltaV2-, deltaV3-, betaF1-). Both patients died a short time after diagnosis. CONCLUSIONS: These observations suggest that at least two forms of nodal gammadelta T-cell NHL may be encountered: one mimicking classical alphabeta T cell NHL, with diffuse pleomorphic cell proliferation, and one displaying sinusoidal neoplastic infiltration suggesting a close relationship with hepatosplenic gammadelta T-cell NHL. PMID- 10692027 TI - Giant lamellar bodies as a feature of pulmonary low-grade MALT lymphomas. AB - AIMS: Giant lamellar bodies (GLBs) are rare pulmonary inclusions, most frequently described in sclerosing haemangiomas. Following a recent report of their presence in a case of pulmonary lymphoma of MALT origin, our aims were to determine their frequency in pulmonary lymphoproliferative disorders, examine their structure and investigate their aetiology further. METHODS AND RESULTS: We reviewed a series of 29 pulmonary lymphomas (23 low-grade, six high-grade) and 18 cases of reactive pulmonary lymphoid hyperplasia. Five of 23 (22%) low-grade lymphomas contained GLBs, 4/4 of which stained for surfactant apoprotein A but not for surfactant apoprotein B. No GLBs were seen in 18 cases of reactive pulmonary lymphoid hyperplasia or six high-grade primary pulmonary lymphomas. Ultrastructural examination revealed concentrically arranged extracellular material forming roughly spherical structures up to 25 microm in diameter. The GLBs were often surrounded by foamy cells and cholesterol clefts, supporting an origin, at least in part, from products of cell breakdown and surfactant degradation. CONCLUSION: These findings support the idea that the presence of lamellar bodies is in part due to stasis of products arising from degradation of surfactant, in association with certain types of chronic pulmonary pathology. Given their absence in reactive pulmonary lymphoid hyperplasia, the presence of GLBs as an epiphenomenon in a pulmonary lymphoid infiltrate should warrant careful investigation with regard to the diagnosis of low-grade MALT lymphoma. PMID- 10692028 TI - Reduced expression of the cell-cycle inhibitor p27Kip1 is associated with progression and lymph node metastasis of gastric carcinoma. AB - AIMS: p27Kip1 (p27), a cyclin-dependent kinase inhibitor, plays an important role as inhibiting the progression of the cell cycle. Decreased expression of p27 is associated with high histological grade and aggressiveness of several human tumours. We aimed to evaluate the role of p27 in the progression and metastasis of gastric carcinoma. METHODS AND RESULTS: We analysed the expression of p27 in 67 primary gastric carcinomas and 31 lymph node metastases by immunohistochemistry. Reduced expression of p27 was found more frequently in advanced gastric cancer (40.9%) than in early gastric cancer (15.6%) (P < 0.001). Decreased p27 expression correlated with large tumour size, high histological grade, lymphatic invasion, advanced stage, deep invasion, lymph node metastasis and recurrence. The expression of p27 showed an inverse correlation with the Ki67 labelling index. There was a significant reduction of p27 expression in metastatic tumour cells in lymph nodes (mean positive cells: 3. 7%) when compared to the corresponding primary gastric carcinomas (mean positive cells: 8.1%) (P = 0.008). CONCLUSIONS: Alterations of p27 expression may play an important role in the progression and metastasis to lymph node of tumour cells in human gastric carcinoma. PMID- 10692029 TI - Epstein-Barr virus and gastric carcinoma in Western patients: comparison of pathological parameters and p53 expression in EBV-positive and negative tumours. AB - AIMS: The presence of Epstein-Barr virus (EBV) was studied in 56 gastric carcinomas from Western patients by in-situ hybridization for EBV-encoded RNAs (EBER). EBV-positive and negative carcinomas were compared for various pathological parameters including p53 overexpression. METHODS AND RESULTS: EBERs transcripts were detected in seven cases overall: four cases of 52 conventional carcinomas (7. 7%) and three cases of four gastric carcinomas with lymphoid stroma (75%). EBER positivity was diffuse in five cases and restricted to a localized area of the tumour in two cases of conventional carcinoma. A monoclonal EBV genomic pattern was demonstrated in the case tested by Southern blot analysis. By immunohistochemical analysis, neither EBV latent or lytic cycle proteins nor C3d/EBV receptor were expressed by neoplastic cells. EBER positivity was significantly correlated with prominent lymphoid reaction (P = 0.0002) which was associated with numerous PS100-positive dendritic cells and with HLA-DR expression by tumour cells (P = 0.03). p53 immunoreactivity in more than 30% of tumour cells was detected in 25 out 49 EBV-negative cases and was absent in EBV positive cases except in one case with focal EBER-positivity. CONCLUSIONS: Focal staining for EBER is an unusual finding in the setting of gastric carcinoma and these results suggest that there might be two types of EBV-associated gastric carcinoma in which the viral infection will play a different role. The presence of a stromal lymphoid reaction which is strongly correlated with EBV positivity, is associated with antigen-presenting ability by HLA-DR-positive tumour cells or abundant dendritic cells. The function of p53 appears preserved in all EBV associated carcinomas except in one case with focal EBER expression whereas the immunohistochemical pattern of p53 is suggestive of a mutational phenomenon in 51% of EBV-negative cases. PMID- 10692030 TI - Delayed appearance of decorin in healing burn scars. AB - AIMS: We have previously shown that hypertrophic scar tissue from burn patients contains abnormally high amounts of the proteoglycans versican and biglycan and reduced amounts of decorin, in comparison with normal dermis or mature scar. The lack of decorin may account for the poor organization of collagen fibrils in the nodular areas of these scars. Decorin has also been reported to neutralize the fibrogenic growth factor TGF-beta1. This study was conducted to monitor the time course of expression of decorin in healing burn wounds by in-situ hybridization to determine whether its absence from hypertrophic scars could result from reduced synthesis. METHODS AND RESULTS: Scar tissue from 19 patients and normal dermis from six patients, was fixed in paraformaldehyde, embedded in paraffin and sectioned. Digoxigenin-labelled cRNA probes were prepared from a plasmid containing a 622-bp insert of human decorin cDNA and used for in-situ hybridization. Total numbers of connective tissue cells and cells positive for decorin mRNA were counted in 10 random fields in the upper (papillary), middle and lower (reticular) one-thirds of the dermis. In all regions the number and percentages of cells with decorin mRNA were low during the first 12 months after injury (eight samples), much higher between 12 and 36 months (seven samples) and low and similar to those in normal skin after 36 months (five samples). The differences between intermediate and early or late stage samples were statistically significant (one-way ANOVA). Immunohistochemistry showed little staining for decorin in early stage samples and much stronger staining in mid stage. Late stage tissue showed intense staining for decorin, almost comparable to that in normal dermis. CONCLUSION: Expression of decorin in burn wounds is suppressed for about 12 months and then increases at a time when resolution of hypertrophic scarring is generally considered to occur. PMID- 10692031 TI - Specific markers for pulmonary tumours. PMID- 10692032 TI - Antiviral protection after DNA vaccination is short lived and not enhanced by CpG DNA. AB - In this study, we investigated the potential of a DNA vaccine expressing the minimal cytotoxic T lymphocyte (CTL) epitope gp33 of the lymphocytic choriomeningitis virus glycoprotein to protect against infection of a non lymphoid organ and compared this to protection against a systemic infection. Furthermore, since immune stimulatory sequences have been shown to augment CTL responses, we examined the capacity of CpG DNA to enhance CTL memory. The data show that DNA vaccination with a gp33-based gene construct induced short-lived gp33-specific CTL which protected against a systemic infection but not against a peripheral infection. Immune stimulatory sequences were incapable of either prolonging CTL memory or promoting protection against infection of a peripheral organ. PMID- 10692033 TI - CpG-oligodeoxynucleotides enhance T-cell receptor-triggered interferon-gamma production and up-regulation of CD69 via induction of antigen-presenting cell derived interferon type I and interleukin-12. AB - Bacterial cytidine-phosphate-guanosine (CpG-DNA) activates antigen-presenting cells (APC) and drives T helper 1 (Th1)-polarized immune responses in the mouse. Claims have been made that CpG-DNA costimulates murine T cells. We examined the direct and indirect effects of CpG-oligodeoxynucleotides (CpG-ODN) on human T cell activation. CpG-ODN failed to costimulate purified human T cells activated with alpha-CD3 or alpha-T-cell receptor (TCR)alphabeta antibodies. In contrast, CpG-ODN sequence-specifically caused increased expression of CD69 on CD4 and CD8 T cells when peripheral blood mononuclear cells (PBMC) were stimulated via alpha CD3. CpG-ODN and alpha-CD3 stimulation synergized to induce interferon-gamma (IFN gamma) in T cells and natural killer (NK) cells, as shown by intracellular fluorescence-activated cell sorter (FACS) staining. These effects of CpG-ODN on human T cells were caused by the release of IFN type I (IFN-I) and interleukin-12 (IL-12) from PBMC. Enhancement of CD69 expression on alpha-CD3-triggered T cells could be reproduced in a coculture transwell system of purified T cells and PBMC, was inhibited by neutralizing antibodies to IFN-I and could be mimicked by adding exogenous IFN-I. Furthermore, neutralization of either IFN-I or IL-12 diminished, and in combination abolished, IFN-gamma production. These findings show that CpG ODN potentiate TCR-triggered activation of human T cells in an APC-dependent manner. PMID- 10692034 TI - Inhibition of immunoglobulin E response to Japanese cedar pollen allergen (Cry j 1) in mice by DNA immunization: different outcomes dependent on the plasmid DNA inoculation method. AB - To develop a new immunotherapy for Japanese cedar (Cryptomeria japonica; CJ) pollinosis, we evaluated the use of DNA immunization by inoculating mice with plasmid DNA encoding Cry j 1 as a CJ pollen major allergen (pCACJ1). Repeated intramuscular (i.m.) inoculation of BALB/c mice with pCACJ1 produced anti-Cry j 1 antibody responses, which were predominately of the immunoglobulin G2a (IgG2a) type. Furthermore, this inoculation suppressed immunoglobulin E (IgE) and IgG1 antibody responses to subsequent alum-precipitated Cry j 1 injections. Splenic T cells isolated from mice inoculated with pCACJ1 i.m. secreted interferon-gamma (IFN-gamma), but not interleukin (IL)-4, in vitro upon stimulation with Cry j 1 as well as with p277-288, a peptide corresponding to the T-cell epitope of Cry j 1. In contrast, inoculation of BALB/c mice with pCACJ1 by gene gun injection caused response predominantly of the IgG1 type, and enhanced production of anti Cry j 1 IgE antibodies to subsequent alum-precipitated Cry j 1 injections. Splenic T cells isolated from pCACJ1-innoculated mice by gene gun injection secreted both IFN-gamma and IL-4 in vitro, upon stimulation with Cry j 1 as well as with p277-288. These findings suggest that i.m. inoculation with pCACJ1 effectively elicits Cry j 1-specific T helper 1 (Th1)-type immune responses, resulting in inhibition of the IgE response to Cry j 1. PMID- 10692035 TI - Vgamma1+ gammadelta T cells play protective roles at an early phase of murine cytomegalovirus infection through production of interferon-gamma. AB - Cytomegalovirus (CMV) causes severe opportunistic infection in immunocompromised hosts. The importance of conventional alphabeta T cells in protection against CMV infection has been well documented. However, the role of the second T-cell population (which express the gammadelta T-cell receptor) in CMV infection is not known. In the present study, we analysed the function and protective role of gammadelta T cells in a murine cytomegalovirus (MCMV) infection model. After intraperitoneal infection with MCMV, the number of gammadelta T cells increased in the liver and peritoneal cavity from day 3, and reached a peak on day 5. The gammadelta T cells showed an activated T-cell phenotype and predominantly expressed Vgamma1, which is known to be expressed by heat-shock protein 65 (hsp 65)-specific gammadelta T cells. Analysis of cytokine expression demonstrated that the MCMV-induced gammadelta T cells expressed interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) but not interleukin-4 (IL-4), implying their participation in the cell-mediated immune response against MCMV. Depletion of gammadelta T cells by anti-T-cell receptor (TCR) gammadelta monoclonal antibody (mAb) treatment resulted in significant increase of virus titre and decrease of IFN-gamma in the liver on day 3 after MCMV infection, which further supports the importance of gammadelta T cells in early protection against infection. Finally, the MCMV-induced gammadelta T cells produced IFN-gamma in vitro in response to hsp 65. Our results suggest that gammadelta T cells participate in early protection against MCMV infection through recognition of hsp 65 and production of IFN-gamma. PMID- 10692036 TI - Bovine tuberculosis: immune responses in the peripheral blood and at the site of active disease. AB - This report describes a comparison of immune responses in the peripheral blood and at the site of active disease in cattle 20 weeks after experimental infection with Mycobacterium bovis. Lymphocyte proliferation, and the production of interferon-gamma (IFN-gamma) and interleukin (IL)-2 were measured in response to tuberculin and a number of mycobacterial antigens, including ESAT-6, MPB64, MPB70, MPB83, hsp 16.1, hsp 65, hsp 70 and the 38 000 MW lipoprotein antigen. The level of transforming growth factor-beta (TGF-beta) was measured following stimulation of cells with tuberculin. Our results suggest little difference in the responses of peripheral blood and lymph node cells to most of the antigens used. However, tuberculin purified protein derivative (PPD) and ESAT-6 elicited stronger responses in the peripheral blood compared with lymph node cells. Investigation of the responding T-cell subpopulations in the peripheral blood showed that both CD4+ and, to a lesser extent, gammadelta T-cell receptor positive (TCR+) T cells contributed to these responses. This is the first report to compare peripheral and local immune responses in bovine tuberculosis. Unlike cases of human tuberculosis where immune activity at the site of disease and anergy in the peripheral blood have been reported, our results suggest that for bovine tuberculosis immune responses occurring in the peripheral blood reflect those at the site of disease. PMID- 10692037 TI - Minor role played by type I tumour necrosis factor receptor in the control of Mycobacterium avium proliferation in infected mice. AB - Control of mycobacterial growth depends on the concerted activity of different cytokines acting in different stages of the development of innate and adaptive immune responses. Tumour necrosis factor-alpha (TNF-alpha) has been shown to play a protective role in Mycobacterium avium infections. Here we assessed the growth of this mycobacterial species in wild-type mice and in mice with a genetically engineered disruption of the type I receptor for TNF-alpha (p55-KO mice). p55-KO mice infected with a low-virulence strain of M. avium exhibited a slightly delayed capacity to eliminate the micro-organisms from the liver as compared with wild-type animals. However, either the growth of this strain in the other organs studied (spleen and lung) or the growth of two other strains of M. avium with intermediate or high virulence, failed to be affected by mutation of the TNF alpha receptor. p55-KO mice were also as protected by the administration of recombinant interleukin-12 as the heterozygous p55 +/- mice. We conclude that signalling through the type I TNF receptor plays a small role in vivo in the induction of mycobacteriostasis during M. avium infection but may improve survival during infection with virulent mycobacteria, independently of the extent of their proliferation. PMID- 10692038 TI - Relationship between disease severity and responses by blood mononuclear cells from patients with rheumatoid arthritis to human heat-shock protein 60. AB - The hypothesis that T-cell responses to the 60 000 MW family of heat-shock proteins (hsp) may be related to the severity of rheumatoid arthritis (RA) was examined. Peripheral blood mononuclear cells (PBMC) from most normal individuals and both early and established RA patients proliferated in vitro in response to human hsp 60 and mycobacterial hsp 65 as well as tetanus toxoid (TT) and mycobacterial purified protein derivative (PPD). PBMC from some patients with established RA gave responses to hsp 60 that were above the normal range and/or peaked earlier than PBMC from normal individuals. The responses of PBMC from established RA to hsp 65, but not PPD or TT, were also higher than those from normal individuals, but the peak responses to all three antigens appeared delayed. Thus a selective increase in responsiveness to hsp 60 develops with disease duration in many RA patients. Six assessments of disease activity and severity were made but apart from rheumatoid factor titre, they were unrelated to the proliferative response. Similarly, disease activity and severity did not differ between those RA patients whose hsp 60 stimulated cells produced interferon-gamma and those who did not, although patients whose hsp 60-stimulated T cells produced interleukin-4 (IL-4) and/or IL-10, appeared to have less disease activity and severity than those who did not. Significant negative correlations were found between IL-10 production by hsp 60-stimulated cells and disease assessments. It is considered that RA is less severe in those patients whose hsp 60-stimulated cells produce T-helper 2 type cytokines. PMID- 10692039 TI - Repeat bacterial challenge in a subcutaneous chamber model results in augmented tumour necrosis factor-alpha and interferon-gamma response, and suppression of interleukin-10. AB - The present study compared the effect of a single or a repeat challenge with the Gram-negative pathogen Porphyromonas gingivalis on the local inflammatory response within subcutaneous chamber model in mice. Subcutaneous chambers were implanted 2 weeks prior to the final challenge. The repeat-challenge (REP) group received two intrachamber bacterial injections 14 days apart, while the single injection group (SIN) received only a single bacterial challenge. Injection of saline was used as the control. The cellular contents of the chamber exudates were used for differential cell counts, and the supernatants were analysed for tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and interleukin (IL)-10 levels. Immunoglobulin G1 (IgG1) and IgG2a levels to P. gingivalis in the exudates were also determined. The results showed that the leucocyte counts increased significantly post-challenge, and the REP group showed the highest number of lymphocytes and neutrophils. Both P. gingivalis-challenged groups exhibited significant increase in TNF-alpha and IL-10 levels at day 1 post challenge. TNF-alpha levels in the chamber exudate were threefold higher in the REP group compared with the SIN group on day 1 post-challenge (P < 0.05). In contrast, IL-10 levels were significantly lower in the REP group 1 day post challenge compared with the SIN group. The REP group had significantly higher levels of IFN-gamma at baseline, and this difference remained significant 1 day post-challenge. Analysis of antibody levels to P. gingivalis showed that while the control and the SIN groups had no anti-P. gingivalis IgG in the chamber exudate during the 7-day study period, the REP group showed high anti-P. gingivalis IgG levels. In addition, the titres of IgG2a were fivefold higher than the IgG1 titres. The results showed that a repeat local challenge with P. gingivalis augmented the proinflammatory cytokines TNF-alpha and IFN-gamma, while inhibiting the accumulation of the anti-inflammatory cytokine IL-10. This shift towards a T helper 1 (Th1)-dominant response was reflected in the relatively high anti-P. gingivalis IgG2a titres in the local inflammatory environment 7 days post challenge. PMID- 10692040 TI - Recessive expression of the H2A-controlled immune response phenotype depends critically on antigen dose. AB - Major histocompatibility complex (MHC) alleles acting as immune response genes are coexpressed in heterozygous individuals and therefore control of immune responses is usually codominant. As an exception to this rule, however, several examples of recessive immune responses have been ascribed to regulatory, e.g. suppressive, interactions. We report here that the recessive phenotype of both antibody and T-cell responses to the mycobacterial 16 000-MW antigen depends critically on a low antigen dose for immunization. On the basis of similar responses in hemi- and heterozygous mice, we suggest that the mechanism of recessive MHC control does not involve regulation by the low-responder allele. We also demonstrated mixed haplotype restriction of peptide recognition for a significant fraction of high-antigen-dose primed T cells. Their paucity under limiting antigen dose conditions may lead to the recessive expression of MHC control. In conclusion, our results suggest that recessive MHC control can be explained as a simple gene dosage effect under conditions where antigen is limiting, without a need for regulatory mechanisms. PMID- 10692041 TI - Human invariant valpha24+ natural killer T cells activated by alpha galactosylceramide (KRN7000) have cytotoxic anti-tumour activity through mechanisms distinct from T cells and natural killer cells. AB - Human Valpha24 + NKT cells, a subpopulation of natural killer cell receptor (NKR P1A) expressing T cells with an invariant T-cell receptor (TCR; Valpha24JalphaQ) are stimulated by the glycolipid, alpha-galactosylceramide (KRN7000), in a CD1d dependent, TCR-mediated fashion. Little is known about Valpha24 + NKT-cell function. The murine counterpart, Valpha14 + NKT cells, appear to have an important role in controlling malignancy. There are no human data examining the role of Valpha24 + NKT cells in controlling human malignancy. We report that Valpha24 + NKT cells have perforin-mediated cytotoxicity against haemopoietic malignancies. Valpha24 TCR, CD1d and alpha-galactosylceramide may all play a role in cytotoxicity but are not absolute requirements. The greatest cytotoxicity was observed against the U937 tumour cell line (95 +/- 5% lysis). THP-1, Molt4, C1R cells and allogeneic mismatched dendritic cells were also sensitive to Valpha24 + NKT cytotoxicity but neither the NK target, K562, nor lymphokine-activated killer sensitive Daudi cells, were sensitive. These results indicate a killing pattern distinct from conventional major histocompatibility complex-restricted T cells, NK cells and other cytotoxic lymphoid cells previously described. We conclude that human Valpha24 + NKT cells have cytotoxic anti-tumour activity against haemopoietic malignancies through effector mechanisms distinct from conventional T cells and NK cells and that their specific stimulator KRN7000 may have therapeutic potential. PMID- 10692042 TI - Increasing the frequency of T-cell precursors specific for a cryptic epitope of hen-egg lysozyme converts it to an immunodominant epitope. AB - Efforts to understand the mechanisms that govern how immunodominant T-cell epitopes are selected from protein antigens have focused mostly on differences in the efficiency of processing and presentation of peptide/major histocompatibility complex (MHC) complexes by antigen-presenting cells, while little attention has been directed at the role of the T-cell repertoire. In this report, the influence of the T-cell repertoire on immunodominance was investigated using transgenic mice that express the beta chain from a T-cell receptor specific for a cryptic Ek restricted epitope of hen-egg lysozyme, HEL85-96. In these mice, the frequency of HEL85-96-specific T-cell precursors is increased 10-20-fold over non-transgenic mice. Transgenic mice respond as well as non-transgenic controls to intact HEL, even though they respond poorly or not at all to a variety of other antigens, including the dominant H-2k restricted epitopes of HEL. Following immunization with native HEL, the only HEL peptide that could recall a response in vitro in the transgenic mice was HEL85-96. Therefore, this normally cryptic epitope is the sole immunodominant epitope in the transgenic mice, and this alteration in immune response is due solely to an increase in the frequency of specific T-cell precursors. An analysis of four additional H-2k restricted cryptic epitopes of HEL suggests that three are similarly limited by T-cell frequency, and that only one is consistent with a defect in efficient antigen presentation. This indicates that there are at least two different types of cryptic epitopes, one in which crypticity is caused by inefficient processing or presentation, and another in which the frequency of specific T-cell progenitors is limiting. PMID- 10692043 TI - In vivo rapid reduction of alloantigen-activated CD8+ mature cytotoxic T cells by inhibitors of acidification of intracellular organelles, prodigiosin 25-C and concanamycin B. AB - Prodigiosin (PrG) 25-C and concanamycin B (CMB) are immunosuppressants that specifically inhibit the induction of cytotoxic T cells (CTL) without affecting the function of B cells and helper T cells in vivo. Both compounds inhibit acidification of intracellular organelles and induce destruction of cytotoxic granules and degradation of perforin in vitro. Here we show that a single intraperitoneal (i.p.) injection of PrG 25-C, and of CMB, into mice eliminates cytotoxic activity 7 days after alloantigen stimulation (when mature CTL activity has been detected in control mice), with minimal effect on the alloantigen specific antibody titre in serum. FK506 did not suppress the cytotoxic activity with this administration schedule. Suppression was accompanied by a decrease in the CD8+ population and in perforin expression of spleen cells induced by alloantigen stimulation. The suppression of CTL activity and decrease in CD8+ cell number was detected as early as 7 hr after the injection of compounds. These results suggest that inhibitors of acidification of intracellular organelles suppress CTL activity in vivo by reducing the number of mature CD8+ CTL. PMID- 10692044 TI - T-cell apoptosis and differential human leucocyte antigen class II expression in human thymus. AB - Relatively little is known of the details of human leucocyte antigen (HLA) expression and thymocyte selection in human thymus. In both humans and mice major histocompatibility complex (MHC) molecules have been described which show a highly restricted thymic expression. Such patterns may offer clues about cellular interactions in thymic selection because transgenic mice with MHC expression targeted to specific thymic sites show altered T-cell receptor (TCR) repertoire selection. We have analysed human thymic HLA class II expression, relating the expression pattern to sites of thymocyte apoptosis. While HLA-DQ is poorly expressed by most peripheral antigen-presenting cells (APC), thymus stains strongly for HLA-DQ as well as for HLA-DR. HLA-DM is abundant in medulla but weakly expressed by cortical cells. Class II expression in Hassall's corpuscles (HC) is unusual in several respects: we have previously shown them to be encircled by HLA-DO+ epithelial cells and here further demonstrate that HC are negative for HLA-DR and HLA-DP, but often positive for HLA-DQ and HLA-DM. Transcriptional control of HLA class II products at this site is thus unlike cells that have previously been studied. Apoptotic thymocytes are restricted to the cortex and the corticomedullary junction. However, a minority of apoptotic cells are visible in the medulla, these being found in the HLA-DQ positive HC. The apoptotic thymocytes in HC can be CD4+ single positive (SP), CD8+ SP or CD4+CD8+ double-positive (DP). This study thus shows that the HC within human thymic medulla are noteworthy both for their unusual hierarchy of HLA class II expression and because they are the only medullary site of thymocyte apoptosis. We propose that HC are a site at which mature thymocytes receive activation/tolerization signals from peptides reprocessed from apoptotic cells. The differential HLA transcriptional control at this site may indicate that specific T-cell subpopulations are affected. PMID- 10692045 TI - Characterization and role in experimental systemic lupus erythematosus of T-cell lines specific to peptides based on complementarity-determining region-1 and complementarity-determining region-3 of a pathogenic anti-DNA monoclonal antibody. AB - Peptides based on the complementarity-determining region 1 (CDR1) and CDR3 of an anti-DNA monoclonal antibody (mAb) carrying the 16/6 idiotype (Id) were shown to induce experimental systemic lupus erythematosus (SLE) in susceptible mouse strains. In the present study, T-cell lines specific to the pCDR1 and pCDR3 peptides were established in BALB/c and in SJL mice, respectively. The T-cell lines were characterized and analysed for their pathogenicity upon administration to syngeneic mouse strains. Both T-cell lines expressed the alphabeta T-cell receptor (TCR) and the CD4+ CD8- phenotype. Additionally, both cell lines secreted interleukin (IL)-4 and IL-10 upon stimulation with their specific peptide, thus belonged to the T helper 2 (Th2) subset. Upon immunization, the pCDR3-specific T-cell line induced experimental SLE in SJL mice. The animals produced high levels of autoimmune anti-DNA and antinuclear protein antibodies, as well as anti-16/6 Id antibodies (Abs). Furthermore, the mice developed clinical manifestations, including leukopenia, proteinuria and accumulation of immune complex deposits in their kidneys. The pCDR1-specific T-cell line failed to induce SLE when injected into BALB/c mice. It is thus suggested that pCDR3 is an immunodominant epitope in experimental SLE and that pCDR3-specific T cells initiate autoimmunity, leading to SLE, probably via epitope spreading. PMID- 10692046 TI - Human B-cell lines constitutively express and secrete interleukin-16. AB - Interleukin-16 (IL-16), produced by activated CD8+ T lymphocytes, is inhibitory to human immunodeficiency virus-1 (HIV-1) replication. In an attempt to determine whether human B cells express and secrete IL-16, a wide panel of B-cell lines derived from patients with acquired immune deficiency syndrome (AIDS)-associated B-cell lymphomas (AABCL) (n = 5) and from non-AABCLs (n = 8) were studied. Using reverse transcription-polymerase chain reaction (RT-PCR) analysis, we were able to observe ubiquitous expression of IL-16 mRNA. Kinetic studies on constitutive mRNA turnover and secretion for IL-16 suggests that the optimum expression is at 24 hr. Interestingly, we report, for the first time, IL-16 secretion by human B cell lines. PMID- 10692047 TI - Activation of bovine B cells via surface immunoglobulin M cross-linking or CD40 ligation results in different B-cell phenotypes. AB - Experiments reported herein demonstrate that activation of bovine B cells via surface immunoglobulin M (sIgM) cross-linking, analogous to T-cell independent (TI-2) antigenic stimulation, results in the expression of CD5. Interestingly, in the presence of CD40 ligand, sIgM-mediated induction of CD5 on B cells was inhibited. These findings indicate that activation of bovine B cells via B-cell receptor (BCR) cross-linking results in a CD5+ B-cell phenotype and that CD40 signalling is inhibitory to this process. Analysis of cytokine mRNA indicates that bovine B cells constitutively express tumour necrosis factor-alpha (TNF alpha) and interleukin (IL)-1beta transcripts in vitro, while IL-10 mRNA expression is induced following sIgM cross-linking. IL-12 p40 transcripts were produced by B cells activated by CD40, but not by BCR, ligation. Analysis of cytokine receptor mRNA indicates that activation through CD40, in the presence or absence of IgM cross-linking, results in increased IL-4 receptor-alpha (IL 4Ralpha), IL-13Ralpha1 and interferon-alpha receptor 1 (IFN-alphaR1) mRNA levels. Overall, these findings suggest that activation of bovine B cells through BCR cross-linking yields an activation phenotype that differs substantially from that of B cells activated through CD40. PMID- 10692048 TI - CD40-CD40 ligand (CD154) engagement is required but may not be sufficient for human T helper 1 cell induction of interleukin-2- or interleukin-15-driven, contact-dependent, interleukin-1beta production by monocytes. AB - To investigate whether antigen-independent, interleukin-2 (IL-2) or IL-15 activation of polarized T helper (Th) cells would result in contact-dependent activation of monocytes, living Th1 and Th2 cell clones were co-cultured with THP 1 cells or fresh peripheral blood monocytes. Under these conditions IL-1beta production was induced almost exclusively by Th1 cells and was dependent on the presence and dose of IL-2 or IL-15, and on cell-cell contact, as demonstrated by double-chamber cultures. Low levels of IL-1 receptor antagonist (IL-1Ra) were induced by Th1 and higher levels by Th2 cells. IL-10 production was similar in Th1/monocyte and Th2/monocyte co-cultures, thus arguing against preferential down regulation of IL-1beta production by anti-inflammatory IL-10 in Th2 co-cultures. In addition, IL-4 and IL-10 neutralization did not result in enhanced IL-1beta production in Th2/monocyte co-cultures. Preferential expression on Th1 cells of CD11b correlated with their capacity to induce IL-1beta production by THP-1 cells in the presence of IL-2 or IL-15, but anti-CD11b monoclonal antibody could not inhibit this activity. Blockade of the CD40-CD40 ligand interaction resulted in inhibition of IL-1beta-inducing capacity while IL-1Ra induction was unaffected, a result previously unknown. This differential effect indicates the selective relevance of CD40-CD40 ligand engagement in inflammatory monocyte responses upon activation by T cells. CD40 ligand expression levels did not differ in Th1 and Th2 cell clones, thus indicating that additional, unidentified molecule(s) preferentially expressed by Th1 cells are involved in their IL-1beta induction capacity. PMID- 10692049 TI - Inhibition of contact sensitivity in human CD4+ transgenic mice by human CD4 specific monoclonal antibodies: CD4+ T-cell depletion is not required. AB - Clenoliximab and keliximab are monkey/human chimeric monoclonal antibodies (mAbs) of the immunoglobulin G4 (IgG4) and IgG1 isotypes, respectively, that recognize the same epitope on human CD4. The two mAbs possess identical idiotypes and exhibit equal affinities for CD4. Upon administration of these mAbs to mice that express a human CD4 transgene, but not mouse CD4 (HuCD4/Tg mice), clenoliximab and keliximab exhibited similar kinetics of binding to CD4, and induced the same degree of CD4 modulation from the cell surface, although only keliximab mediated CD4+ T-cell depletion. Epicutaneous sensitization and challenge of HuCD4/Tg mice with the hapten oxazolone resulted in a contact sensitivity response characterized by tissue swelling, and the presence of interferon-gamma (IFN gamma) and interleukin-4 (IL-4) in the local tissue. Administration of a single 2 mg dose of either clenoliximab or keliximab to HuCD4/Tg mice prior to sensitization significantly reduced post-challenge tissue swelling, and levels of IFN-gamma and IL-4, indicating that CD4+ T-cell depletion is not required for anti-CD4 mAb-mediated inhibition of contact sensitivity. Administration of either mAb prior to challenge failed to inhibit the contact sensitivity response, indicating differential sensitivity of the afferent and efferent phases of the response to inhibition by CD4-specific mAbs. Collectively, these data indicate that CD4 functions as a positive regulatory molecule in the contact sensitivity response. PMID- 10692050 TI - Studies on delayed systemic effects of ultraviolet B radiation on the induction of contact hypersensitivity, 3. Dendritic cells from secondary lymphoid organs are deficient in interleukin-12 production and capacity to promote activation and differentiation of T helper type 1 cells. AB - Ultraviolet-B radiation (UVR) of mouse skin promotes both local and systemic immune aberrations that are thought to be important in the pathogenesis of cutaneous malignancies. Acute, low-dose UVR regimens inhibit the induction of contact hypersensitivity (CH) in genetically susceptible mice by TNF-alpha dependent mechanisms. In addition, these regimens also promote the development of tolerance when hapten is applied to the UVR-exposed site at the completion of the radiation treatment protocol. A third immune abnormality is also observed in mice exposed to acute, low-dose UVR. This abnormality, which develops within 48-72 hr of the completion of the UVR regimen, has been described among antigen-presenting cells within secondary lymphoid organs, including lymph nodes that do not drain the site of irradiation. Dendritic cells (DCs) from lymph nodes and spleens of mice exposed to UVR lack the capacity to induce CH if they are derivatized with hapten and injected intracutaneously into naive mice. The DC defect is related to the production of and systemic dissemination of interleukin-10 (IL-10) by keratinocytes within the epidermis of the UVR-exposed skin. We have now examined the nature of the functional aberration that exists among DCs within the secondary lymphoid organs of UVR-exposed mice by examining the capacity of DCs to express co-stimulatory molecules, and their ability to activate ovalbumin (OVA) specific DO11.10 T-cell receptor transgenic T cells in vitro. Our results indicate that DCs from UVR-exposed mice produced insufficient amounts of IL-12. When pulsed with OVA, these cells were capable of inducing proliferation among DO11.10 T cells in vitro, but the responding cells produced neither IFN-gamma nor IL-10 and IL-4. A similar antigen-presenting cell defect was generated in mice treated with a subcutaneous injection of IL-10. We conclude that acute, low-dose UVR creates an IL-10-dependent functional deficit in DCs in secondary lymphoid organs, and that this defect robs UVR-exposed mice of the capacity to develop CH when hapten is painted epicutaneously. PMID- 10692051 TI - Modulation of CD11C+ splenic dendritic cell functions in murine visceral leishmaniasis: correlation with parasite replication in the spleen. AB - BALB/c mice resolve Leishmania donovani infection in the liver over an 8-12-week period. However, after an initial phase of 2-4 weeks where increases in parasite load are not readily detectable, parasite numbers in the spleen begin to increase reaching maximum levels at 16 weeks post-infection. Thereafter, parasite replication in the spleen is controlled and BALB/c mice maintain this residual parasite load in the spleen for many months, without further increase. We evaluated functions of CD11C+ splenic dendritic cells throughout the course of L. donovani infection in the spleen of BALB/c mice. Unlike the dendritic cell (DC) specific antigen DEC-205, CD11C was not up-regulated on macrophages during visceral leishmaniasis. No appreciable impairment of splenic DC functions was observed when this antigen-presenting cell subset was purified from 30-day post infected mice. Significant impairment in inducing allogeneic mixed lymphocyte reaction (MLR) and presenting L. donovani antigens or keyhole limpet haemocyanin (KLH) to specific T cells was observed with CD11C+ splenic DC purified from 60 day post-infected mice. Functional impairment of splenic DC at 60 days post infection correlated with their reduced surface expression of major histocompatibility complex (MHC) class II molecules, impairment of interleukin-12 (IL-12) production and to their ability to suppress interferon-gamma (IFN-gamma) production by Leishmania antigen-primed T cells. Of interest, the impairment of splenic DC in presenting Leishmania antigens or KLH to specific T cells was corrected at 120 days post-infection, and correlated with their up-regulation of MHC class II expression, IL-12 production, induction of IFN-gamma by Leishmania antigen-primed T cells and the onset of control over splenic parasite replication in vivo. These results indicate that functional integrity of DC may be important in controlling L. donovani infection. PMID- 10692052 TI - Demonstration that platelet-activating factor is capable of activating mast cells and inducing a chemotactic response. AB - Platelet-activating factor (PAF) is generated in a variety of inflammatory conditions in which mast cells accumulate. However, little is known about the ability of PAF to influence mast cell function directly. In this study we examine the ability of PAF to activate mast cells and regulate mast cell chemotaxis. PAF was found to induce intracellular calcium mobilization and chemotactic responses in both murine and human mast cells. PAF induced transient increases in intracellular Ca2+ concentrations with a 50% effective dose of 1 nM and induced significant migratory responses at PAF concentrations of 1 nM to 1 microM in the human leukaemia mast cell line (HMC-1). Using signal transduction inhibitors, both PAF-induced calcium mobilization and migration of mast cells were shown to require activation of pertussis toxin-sensitive G proteins. PAF-induced calcium and chemotactic responses were cross-desensitized by C5a. Together, these data demonstrate that PAF is capable of activating distinct signalling pathways in mast cells associated with calcium mobilization and cell migration; and that PAF may thus contribute to the regulation of mast cell responses and hyperplasia at sites of inflammation. PMID- 10692053 TI - Soluble L-selectin levels in type I diabetes mellitus: a surrogate marker for disease activity? AB - L-selectin (CD62L) is a cell adhesion molecule which plays a key role in the initiation of leucocyte migration from blood vessels to sites of local inflammation. The aim of this study was to investigate T-lymphocyte expression of CD62L antigen and serum levels of soluble L-selectin (sL-selectin) in subjects with clinical and preclinical type I diabetes to determine whether they could provide surrogate markers for disease activity. CD62L selectin expression on memory T lymphocytes was studied by cytometric analysis in 22 patients with newly diagnosed type I diabetes, 20 first-degree relatives of patients with type I diabetes, 14 patients with Graves' disease, and 22 healthy controls. sL-selectin levels were measured by enzyme-linked immunosorbent assay (ELISA) in enlarged groups of subjects in these categories, as well as in patients with long-standing type I diabetes, treated Graves' disease and type II (non-insulin dependent) diabetes. L-selectin levels were also related to islet autoantibodies, human leucocyte antigen (HLA) genotype and L-selectin T668C gene polymorphisms. L selectin expression on memory T lymphocytes was reduced in newly diagnosed diabetes and islet autoantibody positive siblings compared with controls. sL selectin levels were significantly raised in newly diagnosed type I diabetes compared with controls, with intermediate levels in family members, both with and without islet autoantibodies, and in long-standing type I diabetes. Levels were also raised in patients with untreated Graves' disease. Patients with type II diabetes had sL-selectin levels which did not differ from controls. sL-selectin levels correlated with the presence of diabetes-associated HLA alleles in both family members and controls; levels also fell with increasing age in family members. Multiple regression analysis showed that HLA genotype and age were independent determinants of sL-selectin levels. sL-selectin levels are raised at the time of diagnosis of type I diabetes and Graves' disease and appear to be modulated by disease activity, but levels are determined predominantly by HLA associated genetic susceptibility and age. sL-selectin may provide a late marker of autoimmune destruction of islets and sequential measurement may be useful in monitoring disease activity and the effect of interventions preceding type I diabetes. PMID- 10692055 TI - Review article: a therapeutic update on dermatomyositis/polymyositis. PMID- 10692054 TI - Production and functional characterization of a soluble recombinant form of mouse CD59. AB - This report describes the engineering, expression, purification and functional characterization of a soluble recombinant form of murine CD59 (srMoCD59). We report the expression in Chinese hamster ovary (CHO) cells of a modified mouse CD59 cDNA that had been truncated at D-74, resulting in the loss of the glycosylphosphatidyl inositol (GPI) anchor, and containing six additional C terminal histidines. The expressed srMoCD59 was purified from tissue culture supernatant by means of its poly-histidine tag using immobilized metal affinity chromatography. In comparison with CD59 on mouse erythrocytes, the srMoCD59 had a reduced molecular weight (18-20 000 as compared with 20-28 000 for GPI-anchored srMoCD59). The terminal complement inhibitory capacity of this soluble recombinant protein was assessed using two methods: a cobra venom factor (CVF) triggered 'reactive-lysis' system and a C5b-7 site assay. In both assays, srMoCD59 inhibited lysis by the sera from all three species tested in the rank order mouse > rat >> human. The amount of srMoCD59 required to produce 50% inhibition of lysis in the C5b-7 site assay, using purified terminal components to develop lysis, was 10-fold less than that required in the same assay when EDTA serum was used as a source of C8 and C9, or in the CVF reactive lysis system. These data indicate that the presence of serum markedly interfered with the activity of srMoCD59 and have important implications for the use of recombinant soluble CD59 analogues as therapeutic agents in complement-mediated diseases. PMID- 10692056 TI - Review article: endocrinopathies and the skin. PMID- 10692057 TI - Commentary: onychomycosis: an epidemio-etiologic perspective. PMID- 10692058 TI - Epidemiology of pemphigus in Sofia, Bulgaria. A 16-year retrospective study (1980 1995). AB - BACKGROUND: Pemphigus is a disease showing an uneven geographical distribution. In Bulgaria pemphigus has always represented a substantial part of diagnosed bullous diseases, but previous epidemiological data are incomplete. Our purpose was to evaluate retrospectively the incidence and prevalence of pemphigus in the district of Sofia (the capital of Bulgaria; population 1 200 000) for a sixteen year period. METHODS: The files of all the newly registered patients with pemphigus in the City Hospital of Dermatology in Sofia during the period Jan 1 1980 to Dec 31 1995, were collected and analysed with regard to personal statistics, ethnic origin, profession, history of the disease including age and season of onset, symptoms, clinical diagnosis, severity, laboratory findings, associated illnesses, therapy, and cure rate. Special attention was paid to smoking, alcohol abuse, and the presence of triggering factors such as emotional stress, drug intake, underlying diseases, neoplasias, or others. RESULTS: During the 16-year period studied, 74 newly diagnosed cases of pemphigus occurred in the district of Sofia, giving a prevalence of 0.38 per 100 000 inhabitants and a mean incidence of 0.47/100 000/year for the overall population and 0.51/100 000/year for the population aged above 20 years. The most common clinical variant is pemphigus vulgaris, frequently occurring in the fifth-sixth decades. The vast majority of the patients are workers or professionals. CONCLUSIONS: The results of the present retrospective study reveal a relatively high prevalence and incidence of pemphigus in Bulgaria, compared to that encountered in other countries. Our data is similar to that reported from Greece. Whether the Balkan Peninsula represents a focus of population groups with high susceptibility to pemphigus is a problem which could be highlighted by further epidemiological studies in this geographic area. PMID- 10692059 TI - Search for evidence of a Th2 profile in HIV+ patients. AB - BACKGROUND: Hypersensitivity dermatoses are common in human immunodeficiency virus-positive (HIV+) patients, particularly as the disease progresses. Studies have shown that a switch to T-helper 2 (Th2) might represent a turning point in HIV. This study investigated whether increases in the number of skin mast cells, immunoglobulin E (IgE) serum levels, and eosinophilia, involved in the Th2 response in allergic disease, might also be present in HIV+ patients. If so, these alterations might explain one of the mechanisms of skin hypersensitivity in these patients. METHODS: Forty-five skin biopsies from the normal skin of the upper arm of HIV+ patients and 15 controls were included in the study. HIV+ individuals were classified into three equal categories according to their immunologic status: Category I (< 200/microL), Category II (200-499/microL), and Category III (> 500/microL). Anti-tryptase antibody was employed in tissue sections to show mast cells; IgE serum levels and eosinophils in peripheral blood count were investigated; delayed-type hypersensitivity (DTH) skin tests (candidin, trichophytin, and PPD 2U) were evaluated. RESULTS: Normal cutaneous mast cell and eosinophil counts were the same in all categories and in the control group, but increased IgE levels (P < 0. 01) and DTH skin test anergy (P < 0.006) were observed among acquired immunodeficiency syndrome (AIDS) patients. CONCLUSIONS: The density of skin mast cells in HIV infection was not modified in the course of the disease. Mast cells do not seem to be primarily responsible for triggering hypersensitivity dermatoses among AIDS patients, although data in support of the Th2 response, as seen in increased IgE serum levels and DTH anergy, are present. PMID- 10692060 TI - Cytodiagnosis of cutaneous basal and squamous cell carcinoma. AB - OBJECTIVE: We performed a prospective study to evaluate the diagnostic accuracy of cytologic examination in basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs), in order to assess its clinical value. Study design Samples were taken by the "scraping" technique which involves scraping with a scalpel blade directly over the skin tumor surface, smearing the cells onto several glass slides, and fixing them with "citospray." The specimens were stained with the Papanicolaou stain. Punch biopsies were taken to confirm the clinical and cytologic impression. RESULTS: We collected 45 skin tumors in total, clinically presumed to be either BCC (n = 15) or SCC (n = 30). Imprint cytology demonstrated to be of help in the rapid diagnosis of skin tumors. CONCLUSIONS: Cytologic examination is easy to perform, saves time, provides a rapid diagnosis, and can be considered, under experienced hands, reliable in the confirmation of malignant skin tumors. Cytology does not give much information about tumor patterns or subtypes which can be related to aggressive behavior and can be very important in further therapeutic decisions. Therefore, histopathologic confirmation is mandatory before any therapeutic maneuver. PMID- 10692061 TI - Comparison of histopathologic and clinical evaluations of pathergy test in Behcet's disease. AB - BACKGROUND: The pathergy test, an important test in the diagnosis of Behcet's disease, is currently applied with disposable/sharp needles and evaluated only clinically (no histopathologic evaluation). In this study, the usefulness of the pathergy test conducted intradermally and intravenously with disposable/sharp needles in the diagnosis and determination of the activation of the disease is studied in comparison with the test conducted with nondisposable/blunt needles. In addition, histopathologic evaluation of the pathergy test is compared with the clinical evaluation. METHODS: The study group consists of 43 Behcet's disease patients together with 15 patients with dermatosis as the control group. The pathergy test was applied to the Behcet's disease patients and the control group intradermally and intravenously with disposable/sharp and nondisposable/blunt needles. RESULTS: The results of the pathergy test on the patients and the control group were evaluated clinically and histopathologically. CONCLUSIONS: Clinical evaluation of the pathergy test conducted intradermally with nondisposable/blunt needles is sufficient for both the diagnosis and determination of the activation of Behcet's disease. Histopathologic evaluation of the test is not found to be more sensitive than the clinical evaluation. PMID- 10692062 TI - Lichenoid and granulomatous dermatitis. AB - BACKGROUND: The prototypic lichenoid eruptions, lichen planus (LP), lichenoid drug eruptions, secondary syphilis, and collagen vascular disease, are defined histologically by a band-like lymphocytic infiltrate in close apposition to the epidermis. We describe a novel form of lichenoid dermatitis with a granulomatous component. DESIGN: Skin biopsies from 40 patients demonstrating a band-like lymphocytic infiltrate with concomitant granulomatous inflammation were encountered over 4 years. Clinicians were contacted to elucidate underlying triggers and medical illnesses. RESULTS: A lichenoid dermatitis, a linear eruption, vasculitis, annular erythema, and erythroderma were among the clinical presentations. A drug-based etiology was implicated in 14 cases: the drugs included antibiotics, lipid-lowering agents, anti-inflammatory drugs, antihistamines, hydroxychloroquine sulfate, and angiotensin-converting enzyme inhibitors. Over one-third of patients with drug-related eruptions had other medical illnesses associated with cutaneous granulomatous inflammation, namely rheumatoid arthritis (RA), Crohn's disease, hepatitis C, diabetes mellitus, and thyroiditis. A microbial trigger was implicated in 12 patients in the context of infective id reactions to herpes zoster, Epstein-Barr virus (EBV), or streptococci, or active infections by Mycobacterium tuberculosis, M. leprae, fungi, and spirochetes. The remainder had hepatobiliary disease and RA without obvious exogenous triggers, cutaneous T-cell lymphoma (CTCL), and idiopathic lichenoid eruptions (i.e. LP, lichen nitidus, and lichen striatus). One patient with LP had underlying multicentric reticulohistiocytosis. The histiocytic infiltrate assumed one or more of five light microscopic patterns: (i) superficially disposed loose histiocytic aggregates; (ii) cohesive granulomata within zones of band-like lymphocytic infiltration with or without deeper dermal extension; (iii) a diffuse interstitial pattern; (iv) scattered singly disposed giant cells; and (v) granulomatous vasculitis. Additional features included lymphocytic eccrine hidradenitis in those patients with drug reactions, hepatobiliary disease, and antecedent viral illnesses, tissue eosinophilia and erythrocyte extravasation in drug hypersensitivity, granulomatous vasculitis in patients with microbial triggers, drug hypersensitivity or RA, and lymphoid atypia in lesions of CTCL or drug hypersensitivity. CONCLUSIONS: The cutaneous lichenoid and granulomatous reaction may reflect hepatobiliary disease, endocrinopathy, RA, Crohn's disease, infection, or a drug reaction. One-fifth of cases represent idiopathic lichenoid disorders. Lymphoproliferative disease or pseudolymphomatous drug reactions must be considered in those cases showing lymphoid atypia. PMID- 10692063 TI - Oral lichen planus: different clinical features in HCV-positive and HCV-negative patients. AB - BACKGROUND: Hepatitis C virus (HCV) infection induces variable dermatologic manifestations. OBJECTIVE: To determine whether differences exist in the clinical features and behavior of oral lichen planus (OLP) between HCV-positive (HCV+ve) and HCV-negative (HCV-ve) patients. METHODS: Two hundred and sixty three patients (156 women and 107 men), with a mean age of 55.5 years, with OLP (76 HCV+ve and 187 HCV-ve) were clinically evaluated. Previously, all local factors that could modify the clinical characteristics were removed and were monitored carefully following morphology. RESULTS: In both groups, the prevalent clinical form of OLP was the mixed form (33.1% in HCV-ve and 35.5% in HCV+ve patients), in which reticular-plaque lesions coexist with atrophic-erosive ones. The reticular form was more frequent in HCV+ve (25%) than in HCV-ve (18. 7%) patients, whereas plaque lesions were more prevalent in HCV-ve (15.5%) than in HCV+ve (5.2%) patients (P < 0.01, chi-squared test). There were no significant differences in the frequency of erosive (27.2% in HCV-ve and 27.6% in HCV+ve) and atrophic (5.3% in HCV-ve and 5.2% in HCV+ve) forms between the two groups. CONCLUSIONS: Our findings show that there were statistically significant differences between OLP HCV-ve and OLP-HCV+ve groups for reticular and plaque clinical forms. These findings underline the importance of liver examination in all OLP patients, including cases with mild, asymptomatic keratotic forms of the disease. PMID- 10692064 TI - Dermatomyositis presenting as panniculitis. PMID- 10692065 TI - T-cell chronic lymphocytic leukemia mimicking dermatomyositis. PMID- 10692066 TI - Sympathectomy-induced ichthyosis-like eruption. PMID- 10692067 TI - Recessive dystrophic epidermolysis bullosa complicated with nephrotic syndrome due to secondary amyloidosis. PMID- 10692068 TI - An epidemic of porphyria cutanea tarda? PMID- 10692069 TI - Pityriasis lichenoides-like exanthem and primary infection by Epstein-Barr virus. PMID- 10692070 TI - Effects of environmental conditions on microbial proteolysis in a pork myofibril model system. AB - A number of bacterial strains used for meat fermentations were screened for proteolytic activity. A strain of Micrococcus which was found to be proteolytic was evaluated for the effects of environmental conditions on its proteolytic activity against pork myofibrillar proteins using response surface methodology. Three strains of micrococci were also tested for the ability to produce free amino acids from pork myofibrils. Analysis of the effects of environmental conditions showed that proteolytic activity would be minimal under conditions normally found in fermented sausages, thereby suggesting that proteolysis in these products is largely due to endogenous meat enzymes. The three strains of micrococci were shown to produce free amino acids from pork myofibrils, thereby demonstrating the presence of peptidase activity in these strains. PMID- 10692071 TI - Influence of water activity and nutrients on growth and production of squalestatin S1 by a Phoma sp. AB - This study investigated the effects of temperature, nutrient status and water activity (aW) on the production of squalestatin S1 by a Phoma sp. The fungus was grown on malt extract (MEA), wheat extract (WEA), oat extract (OEA) and oil seed rape extract (OSREA) agars at 15, 20 and 25 degrees C and 0.998, 0.995, 0.990, 0.980 and 0.960 aW levels. The growth rate and secondary metabolite formation were followed over a total of 30 d. The maximum growth rate was observed at 25 degrees C and 0.998-0.990 aW for all media types, which was significantly reduced (P = 0.05) for most media at 0.96 aw. The growth rate was greatest for WEA and OEA but the growth form was an effuse exploitative type compared with the dense assimilative type on the richer MEA. The lipid-based OSREA appeared to be a poor growth substrate for this fungus. In contrast to the growth rate data, squalestatin S1 production was maximal for all media types at slightly reduced aw in the range 0.990-0.980. There was greater production of the secondary metabolite under significant water stress (0.960 aW) compared with that with freely available water (0.998 aW). Maximum production was observed in WEA. Production began earlier in WEA and OEA compared with MEA. Squalestatin S1 production was not significantly affected by incubation temperature (P = 0.05). This study has shown that nutritionally depleted substrates may be usefully employed in the production of squalestatin S1 and perhaps also for other secondary metabolites. PMID- 10692072 TI - Diversity among lactococci isolated from ewes' raw milk and cheese. AB - The technological and genetic characteristics of lactococci present in ewes' raw milk and 1-d-old ewes' raw milk cheeses sampled over a 1-year period were investigated. The proportion of lactic acid bacteria isolates from milk samples able to decrease milk pH by more than 1.25 units after 6 h incubation at 30 degrees C reached 14.5% in spring vs 10.7% in summer, 8.3% in autumn and 3.0% in winter. In 1-d-old cheese samples, the proportion of lactic acid bacteria able to lower milk pH by more than 1.25 units increased up to 32.3% in spring vs 23.4% in summer, 8.0% in autumn and 10.3% in winter. Fast acid-producing lactic acid bacteria mainly belonged to the genus Lactococcus. Using polymerase chain reaction protocols, fast acid-producing lactococci were grouped as 61 Lactococcus lactis subsp. lactis, 13 L. lactis subsp. cremoris and 14 L. lactis subsp. lactis biovar diacetylactis. Randomly amplified polymorphic DNA (RAPD) fingerprinting of fast acid-producing lactococci, using two primers, resulted in 21 different RAPD patterns for L. lactis subsp. lactis isolates, nine RAPD patterns for L. lactis subsp. cremoris isolates and three RAPD patterns for L. lactis subsp. lactis biovar diacetylactis isolates. Up to 19 different RAPD patterns were found for L. lactis isolates from cheeses made in a particular month. PMID- 10692073 TI - Response to NaCl of nitrate assimilation and nitrogenase activity of the cyanobacterium Anabaena sp. PCC 7120 and its mutants. AB - The presence of NaCl in the nutrient solution promoted nitrate uptake in parent Anabaena sp. PCC 7120, mutants SP7 (defective in nitrate reductase activity) and SP17 (partially defective in nitrate reductase activity), but not in the mutant SP9 (defective in nitrate transport and reduction). Nitrate reductase activity of the parent and mutant SP17 increased with increasing concentration of nitrate in saline medium, while mutants SP7 and SP9 did not respond to the altered salinity. Although Na+ was not required for nitrate reductase activity, its presence in the nutrient solution enhanced nitrate reduction. Complete removal of Na+ from the nutrient solution markedly reduced nitrogenase activity in all the strains, while raising the concentration of NaCl to 50 mmol l-1 or above, was equally toxic to nitrogenase activity. External NaCl at 200 mmol l-1 brought down the nitrogenase activity to the same residual level as observed without Na+. PMID- 10692074 TI - Evaluation of different PCR-based DNA fingerprinting techniques for assessing the genetic variability of isolates of the fungus Epicoccum nigrum. AB - Thirty-six strains of the fungus Epicoccum nigrum, isolated from different substrata and ecosystems of Europe, America and Africa, were analysed using 14 molecular markers included in 5 different genetic fingerprinting techniques: AP PCR, tDNA-PCR, microsatellite-primed PCR, ARDRA and AFLP. All of the techniques used were able to differentiate the isolates, showing a high genetic diversity within the species. However, the different techniques detected different levels of similarity among the strains; ARDRA shows the most homogeneous results whilst AP-PCR shows the most heterogeneous. The similarity indices achieved for each strain were compared for the different techniques. The distribution obtained by microsatellite-primed PCR was similar to those shown by AP-PCR techniques. tDNA PCR and AFLP rendered similar distributions, and ARDRA showed remarkably different results from the other techniques. The results also reveal the lack of an overall correlation between geographical or ecological origin of the isolates and their genotypes. PMID- 10692075 TI - Studies on strong and weak killer phenotypes of wine yeasts: production, activity of toxin in must, and its effect in mixed culture fermentation. AB - Two different killer phenotypes were detected among K+ (killer) yeasts isolated from spontaneous wine fermentations using a plate bioassay. The two phenotypes differed in their degree of killer activity, and were designated as SK+(strong killer) and WK+(weak killer). Strains showing either phenotype were assayed for expression of killer activity under different growth conditions. Growth in must negatively affected expression of the killer activity of both phenotypes. The supernatant fluids from must cultures showed a lower killing effect than those from yeast phosphate dextrose broth (YPDB) cultures. The ability of the two K+ phenotypes to prevail on K-sensitive yeasts was studied in mixed-culture fermentation experiments. Under these conditions, only strains showing SK+ phenotype were able to prevail on the K-sensitive yeasts. These results suggest that the K+ phenotype could play a relevant role in spontaneous fermentations provided that the strain exhibits an SK+ phenotype, and that the latter phenotype should be preferred when selected K + strains are to be used as fermentation starters. PMID- 10692076 TI - Cluster organization of the genes of Streptomyces pristinaespiralis involved in pristinamycin biosynthesis and resistance elucidated by pulsed-field gel electrophoresis. AB - Streptomyces pristinaespiralis synthesizes pristinamycin, a member of the streptogramin antibiotic family which consists of a mixture of two types of chemically unrelated compounds named pristinamycins I and pristinamycins II. In order to estimate the size of the Strep. pristinaespiralis chromosome and to elucidate the organization of the pristinamycin biosynthetic and resistance genes already identified, it was decided to use the pulsed-field gel electrophoresis technique. Results indicate that the Strep. pristinaespiralis chromosome is linear and about 7580 kb, as previously shown for several other Streptomyces species. By hybridization, it could be shown that the biosynthetic and resistance genes for pristinamycins I and pristinamycins II, except for the multidrug resistance gene ptr, are interspersed and seem to be organized as a single large cluster, covering less than 200 kb corresponding to 2.6% of the total size of the chromosome. The consequences and significance of such a genetic organization are discussed. PMID- 10692077 TI - Rapid detection and quantification of yeast species during spontaneous wine fermentation by PCR-RFLP analysis of the rDNA ITS region. AB - PCR-RFLP analysis of the rDNA-ITS (internal transcribed spacer) region was applied to 174 yeast strains belonging to 30 species of oenological significance and including 27 type strains in order to define a rapid identification protocol for yeast colonies. DraI-or HaeIII-PCR-RFLP patterns were species-specific with the exception of teleomorphic and anamorphic forms. An improved protocol taking about 30 h was used for the detection and quantification of yeast species occurring in the course of a spontaneous wine fermentation at industrial level. Wine samples were taken and plated daily on an agar medium and the developed colonies were analysed by PCR-RFLP after 24 h of incubation. A representative sample of these colonies was also identified by traditional methods. Both procedures gave identical results. However, PCR-RFLP analysis allowed a more precise enumeration of the yeast populations, proving to be a reliable and simple method for monitoring the development of the yeast community throughout wine fermentation. PMID- 10692078 TI - 21st Century: Review: Ageing and medicine. AB - Throughout the world, populations are ageing. The response of the health services needs to be based on a knowledge of the nature of human ageing and the principles of rational health care for older people. Ageing comes about from interactions between intrinsic (genetic) and extrinsic (environment and lifestyle) factors. Health care has to be responsive to the general needs of older people, but also to recognize the heterogeneity produced by different rates and patterns of individual ageing. There are now real possibilities of improving the course of human ageing through modulation of both intrinsic and extrinsic processes. There is also a need to adapt social institutions to what is a permanent change in demography. PMID- 10692079 TI - Molecular aspects of the inherited porphyrias. AB - The porphyrias are diseases due to marked deficiencies of enzymes of the haem biosynthetic pathway (Fig. 1). Except for the first enzyme of the pathway, delta aminolevulinate synthase (ALAS), deficiencies in seven other enzymes are associated with the various forms of porphyria (Fig. 2). Porphyrias can be classified as either hepatic or erythroid, depending on the major site of production of porphyrins or their precursors. The pathogenesis of all inherited porphyrias has now been defined at the molecular level, and it is clear that there is a great deal of genetic heterogeneity in each porphyria [1]. PMID- 10692080 TI - Cortisol axis abnormalities early after stroke--relationships to cytokines and leptin. AB - OBJECTIVE: To assess the relationships between circulating levels of proinflammatory cytokines and adrenocortical hormones and leptin early after stroke. DESIGN: Blood samples were collected four times daily the first two days after stroke, twice daily the next 4 days and four times at day 7. Cognitive function and functional outcome was measured at admittance and at day 7. SETTING: Consecutive inclusion of patients admitted to the stroke unit at Umea University Hospital. SUBJECTS: Eight men and 4 women with acute stroke and 10 healthy volunteers. MAIN OUTCOME MEASURES: Levels and diurnal variations of plasma proinflammatory cytokines interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha), serum adrenocortical hormones (cortisol and DHEA) and leptin, and MMSE, SSS, and ADL scores. RESULTS: A significant correlation was present between IL-6 and cortisol levels the first two days after stroke (P < 0.05). In patients with a disturbed diurnal rhythm of cortisol, cortisol and leptin levels were increased (68% and 81% increase, respectively), whilst DHEA levels were unaltered. Half of the patients displayed an abnormal diurnal rhythmicity of leptin at the end of the week. Median TNF-alpha levels for the first two days after stroke also correlated to median leptin levels at the end of the week (P < 0.05). Median IL-6 levels correlated to severity of paresis on days 1 and 7 and to MMSE scores on day 7 (P < 0.05). CONCLUSIONS: Neuroendocrine disturbances are common and often profound early after stroke. Cytokines seem to be important modulators of these disturbances, including diurnal rhythmicity of cortisol and leptin. PMID- 10692081 TI - The hypothalamic-pituitary-adrenal axis activity as a predictor of cardiovascular disease, type 2 diabetes and stroke. AB - OBJECTIVES: The hypothalamic-pituitary-adrenal (HPA) axis, the mediator of cortisol, plays a central role in the homeostatic processes. In this study, we addressed the potential impact of HPA axis activity on established anthropometric, metabolic and haemodynamic risk factors for cardiovascular disease (CVD), type 2 diabetes mellitus and stroke. DESIGN: A cross-sectional study. SUBJECTS: A subgroup of 284 men from a population sample of 1040 at the age of 51 years. MAIN OUTCOME MEASURES: Anthropometric measurements included body mass index (BMI, kg m-2), waist/hip circumference ratio (WHR) and abdominal sagittal diameter (D). Overnight fasting values of blood glucose, serum insulin, triglycerides, total, low (LDL) and high density (HDL) lipoprotein cholesterol, as well as resting heart rate and blood pressure, were also determined. By using repeated diurnal salivary cortisol measurements during everyday conditions, methods were developed to characterize the status of the HPA axis, and set in relation to the anthropometric, metabolic and haemodynamic measurements. RESULTS: In bivariate analyses, risk factors intercorrelated in clusters of anthropometric (BMI, WHR, D), metabolic (insulin, glucose and their ratio, triglycerides, cholesterol [total and LDL], HDL cholesterol [negative]) and haemodynamic (systolic and diastolic blood pressure and heart rate) measurements. This was also the case in the two-dimensional scaling analysis, where, however, HDL separated out. A normal HPA axis status, characterized by high variability and morning cortisol values, as well as a clear response to a standardized lunch and dexamethasone suppression test, was then introduced by a statistical weighting procedure. This did not essentially change the results of either the bivariate correlation matrix or the two-dimensional scaling analysis. A similar introduction of a pathological HPA axis, characterized by low variability and morning cortisol values, a poor lunch-induced cortisol response and a blunted dexamethasone suppression of cortisol, changed the results markedly. Now strong and consistent correlations were found not only within but also between different clusters of risk factors, which also congregated into one distinct cluster, again except for HDL cholesterol. CONCLUSIONS: These results disclose the prospect of an overriding function of a pathological HPA axis on other, established risk factors for CVD, type 2 diabetes and stroke. Its close association to HPA axis dysfunction may explain the previously reported powerful risk indication of abdominal obesity for the diseases mentioned. The HPA axis abnormality has been reported to be a characteristic consequence of frequently repeated or chronic environmental stress challenges. PMID- 10692082 TI - Independent effects of obesity and cortisol in predicting cardiovascular risk factors in men and women. AB - OBJECTIVES: Recent data suggest that higher plasma cortisol may be associated with hypertension and insulin resistance in otherwise healthy men, as it is in Cushing's syndrome. However, obesity in women is associated with lower plasma cortisol concentrations. This study sought to establish whether plasma cortisol is associated with cardiovascular risk factors in women as it is in men, and whether these relationships in either sex are confounded by obesity. DESIGN: A population-based cross-sectional study. SETTING: The MONICA study in northern Sweden. SUBJECTS: From a target cohort of 2500, 1921 subjects took part and 226 were randomly selected because they attended between 07.00 and 09.00 h after an overnight fast. A 75 g oral glucose tolerance test was performed and blood sampled at baseline and 2 h after glucose. RESULTS: Plasma cortisol was lower in relatively obese subjects: in men, this was observed only in the 2 h sample (r = 0.23, P = 0.02) and in women only in the fasting sample (r = -0.26, P < 0.01). Simple regression analysis did not identify relationships between plasma cortisol and blood pressure, serum lipids, fasting insulin or glucose tolerance. However, after adjusting for the effect of obesity by multiple regression, higher plasma cortisol was independently associated with higher diastolic blood pressure in men (r = 0.21, P = 0.04) but not in women, and higher fasting serum triglyceride levels in women (r = 0.28, P < 0. 001) but not in men. CONCLUSIONS: Increasing obesity and plasma cortisol concentrations make independent and sex-specific contributions to variations in blood pressure and aspects of the insulin resistance syndrome. Adverse cardiovascular risk is greatest in those with the combination of obesity and failure to downregulate plasma cortisol levels. PMID- 10692083 TI - Tissue-type plasminogen activator and C-reactive protein in acute coronary heart disease. A nested case-control study. AB - OBJECTIVES: To study the importance of inflammation and fibrinolysis for evolution of ischaemic heart disease in a cohort of initially healthy subjects. DESIGN: Nested case-control study. Follow-up periods 7-15 years. SUBJECTS: Included in the study were 133 cases with coronary heart disease and 258 controls. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Subjects with ischaemic heart disease identified in 1991 by the Danish National Hospital Register. Protein concentration of C-reactive protein (CRP) and tissue-type plasminogen activator (t-PA) were measured with ELISA methods in stored serum samples. RESULTS: CRP and t-PA concentrations were both significantly higher in cases than in controls (P < 0.001 and P < 0. 001). This difference between cases and controls for CRP and t-PA was present in both men (CRP: P = 0.022; t-PA: P = 0.001) and women (CRP: P = 0.013; t-PA: P = 0.005) and it was present in both the 7-9 years follow-up cohort (CRP: P = 0.014; t-PA: P = 0.001) and the 15 years follow-up cohort (CRP: P = 0.027; t-PA: P = 0.012). The best predictor of CRP was t-PA, whilst the best predictor of t-PA was triglycerides. In a logistic regression analysis model, t-PA still came out as independent predictor of coronary heart disease, whilst such a significance disappeared for CRP. With the use of ROC curves we determined that AUC for t-PA was 0.62, and for CRP 0.59, indicating that none of these two analytes has a high prognostic power in predicting future coronary events in an initially healthy population. CONCLUSION: We conclude that moderate increases in serum concentrations of CRP and t-PA are present for up to 15 years before the presence of clinical overt coronary heart disease; that a low-grade inflammation is determined by other risk factors and that t-PA is an independent risk factor for evolution of coronary heart disease. PMID- 10692084 TI - Helicobacter pylori CagA seropositivity does not influence inflammatory parameters, lipid concentrations and haemostatic factors in healthy individuals. AB - BACKGROUND: H. pylori CagA seropositivity has been recently associated with ischaemic heart disease. OBJECTIVE: To evaluate whether H. pylori virulence has any effect on certain circulating coagulation factors and on markers of systemic inflammation in healthy individuals. DESIGN: Prospective cohort study. SETTING: Haematology and gastroenterology unit at a university teaching hospital. SUBJECTS: A total of 494 consecutive asymptomatic blood donors attending a blood bank. MEASUREMENTS: Blood analysis for haemostatic factors, lipids concentrations, inflammatory parameters as well as determination of anti H. pylori IgG and CagA reactivity by ELISA assayes. RESULTS: The overall prevalence of H. pylori infection was 53%; 56% of H. pylori positive sera expressed CagA reactivity. CagA seropositive subjects did not differ significantly from CagA negative or H. pylori negative subjects in values for lipids, haemostatic factors, or inflammatory parameters. CONCLUSIONS: CagA seropositivity is not associated with increased systemic inflammation or with raised concentrations of haemostatic factors - predictors of ischaemic heart disease - in healthy individuals. PMID- 10692085 TI - Coronary artery disease and arterial hypertension: clinical, angiographic and follow-up data. AB - OBJECTIVES: To evaluate how the presence of arterial hypertension affects coronary atherosclerosis and prognosis in patients with, or at high risk of, ischaemic heart disease. DESIGN: Retrospective analysis of clinical records and follow-up data. SETTINGS: Single referral centre for ischaemic heart disease. SUBJECTS: All consecutive patients (n = 1700, 38% with hypertension) undergoing coronary angiography for the evaluation of ischaemic heart disease during 1983 92. RESULTS: On angiography, the likelihood of having three-vessel disease was higher amongst hypertensives (odds ratio = 1.41; 95% confidence interval [CI] = 1.08-1.85) after adjustment for age, sex, and angina symptoms. The sum of all visible stenoses (an index of overall atherosclerotic involvement) was 19% higher in hypertensives (262 +/- 204 vs. 220 +/- 194 units, P < 0.005). By multivariate analysis, the presence of hypertension made a modest (+ 28 units), albeit statistically significant, independent contribution to the total atherosclerosis score. On follow-up (median = 96 months), cardiovascular mortality was slightly higher in the hypertensive patients than in the normotensive group (P < 0.05 in a Kaplan-Meier analysis), but a proportional hazard analysis adjusting for age and gender showed no significant independent contribution of hypertension. Hypertensive patients, however, remained at higher risk of non-fatal myocardial infarction following discharge (adjusted odds ratio = 1.21, 95% CI = 1.03-1.46; P < 0.05). CONCLUSIONS: In this referral population, hypertension is a risk factor for presence of three-vessel disease. Distribution, severity and extension of coronary stenosis are similar to those of normotensive patients, and prognosis is only marginally affected. PMID- 10692086 TI - Heart rate and mortality. AB - OBJECTIVES: Increased heart rate has shown to be associated with risk of mortality from cardiovascular diseases in some studies, but not in others. Increased heart rate has also been linked to causes of death other than cardiovascular. To clarify the role of heart rate as a predictor of death we studied its predictive value in a large population study. DESIGN: A prospective population study with a follow-up time of 23 years. SUBJECTS AND METHODS: The study population comprised 5598 men and 5119 women 30-59 years of age on entry. Heart rate was measured from resting ECGs. MAIN OUTCOME MEASURE: Mortality from specified causes. RESULTS: A total of 1848 men and 840 women died during the follow-up period. Increased heart rate was significantly associated with death from all causes, cardiovascular causes, and natural noncardiovascular, nonmalignant causes of death. Increased heart rate was associated with death from cancer in men with heart disease but not in men without heart disease on entry into the study. The increase in cardiovascular mortality with high heart rate was explained by the close association between heart rate and blood pressure. Adjustment for risk factors did not alter the significance of the association between increased heart rate and mortality from noncardiovascular causes. CONCLUSIONS: High heart rate is simple to observe clinically and a significant if nonspecific predictor of mortality. Increased risk of mortality from cardiovascular diseases can be explained by association with high blood pressure. The increased mortality risk associated with high heart rate related mainly to a group of diseases of noncardiovascular or nonmalignant origin. PMID- 10692087 TI - Psychosocial variables in relation to various risk factors in patients with stable angina pectoris. AB - OBJECTIVES: To evaluate relationships between psychosocial variables and common risk factors such as age, concomitant diseases (hypertension, diabetes mellitus, myocardial infarction, heart failure) and smoking habits in patients with stable angina pectoris. SETTING: University Hospital. SUBJECTS: Participants in the Angina Prognosis Study in Stockholm (APSIS), which comprised 809 patients (248 females) <70 years of age, with chronic stable angina pectoris, of whom 767 (236 females) participated in the present report. Patients with angina pectoris occurring only at rest constituted one group, patients with angina pectoris on effort with or without angina at rest were stratified according to signs of marked ischaemia on exercise and/or clinical signs of heart failure. METHODS: Psychosomatic symptoms, job strain, Type-A behaviour, sleep disturbances and overall life satisfaction were evaluated by a structured interview, which also included questions regarding how the patients usually felt, and health related problems, according to a standardized check-list. RESULTS: Age correlated with several psychosomatic symptoms and tendency to worry. When adjusted for age and sex, patients with previous myocardial infarction and heart failure described more psychosomatic symptoms, but worried less about the future than patients without these diseases. In the group with angina pectoris at rest only there were fewer smokers than amongst other groups, regardless of risk stratification. CONCLUSIONS: Smoking habits and concomitant diseases influence psychosocial variables in patients with stable angina pectoris. The severity of angina pectoris does not seem to relate to life satisfaction and attitudes towards the future. PMID- 10692088 TI - Clinical assessment of suspected deep vein thrombosis: comparison between a score and empirical assessment. AB - OBJECTIVES: To assess the accuracy and agreement of two methods of clinical evaluation: a formal score based on a number of items of fixed value (the so called Wells' score), and an empirical assessment based on a predefined list of items that can be weighted individually. Clinical probability is essential to manage suspected deep vein thrombosis (DVT) and should be assessed before any diagnostic test. DESIGN: An open, nonrandomised, one-centre study. SETTING: One centre in Switzerland (a university hospital delivering primary-tertiary care). SUBJECTS: Two hundred and seventy outpatients with a prevalence of DVT of 21.1% (final diagnosis), out of an initial population of 328 patients, of which 52 had to be excluded because of a history of DVT (score not applicable) or because of insufficient clinical information (n = 6). RESULTS: Agreement between the two assessment tools was poor (kappa value of 0.32), but accuracy was excellent, with a prevalence of DVT of 1.3%, 18.1%, and 100% (empirical assessment), and 3.2%, 19.4%, and 73.9% (Wells' score), for a low, intermediate or high clinical probability estimate, respectively. The main differences between the two methods were that (i) the empirical method performed slightly better in categorizing patients in the high probability class, and (ii) Wells' score categorized more patients in the low probability class. When applied to two validated diagnostic strategies, the empirical assessment required slightly fewer phlebograms in both strategies, and Wells' score required fewer repeat ultrasonograms (in the strategy that requires this procedure). CONCLUSIONS: Clinical probability assessment can be done with a similar accuracy either empirically or using a score. Institutions should incorporate clinical probability assessment with either method depending upon their diagnostic strategy for suspected DVT. PMID- 10692089 TI - Silent cerebral infarction: a potential risk for pneumonia in the elderly. AB - OBJECTIVE: To determine whether patients who have silent cerebral infarction are more likely to develop pneumonia than are controls without silent cerebral infarction. DESIGN: We examined 269 community-residing participants of the senior day-care centre without history of previous stroke, and then followed them over a two-year period to assess pneumonia. On the basis of computerized tomography scans, they were divided into two groups: no infarction (n = 102) and cerebral hemispheric infarction (n = 167). Cerebral infarcts were further divided into deep and superficial infarcts. RESULTS: The incidence of pneumonia was significantly higher in subjects with silent cerebral infarction (19.8%) than in controls (4. 9%) (odds ratio, 4.67 [95% CI, 1.87-11.67]; P < 0.01). Deep infarcts were more closely associated with the incidence of pneumonia (29.1%) than superficial infarcts (7.6%) (odds ratio, 5.00 [CI, 1.91-13.08]; P < 0.01). CONCLUSIONS: Elderly subjects with silent cerebral infarction were more likely to develop pneumonia than were controls without silent cerebral infarction. Amongst hemispheric silent cerebral infarcts, those located in the deep brain structures may be an important predictor of the development of pneumonia. PMID- 10692090 TI - Commonly recommended daily intake of vitamin D is not sufficient if sunlight exposure is limited. AB - OBJECTIVES: Sunlight exposure of the skin is known to be the most important source of vitamin D. The aims of this study were: (i) to estimate vitamin D status amongst sunlight-deprived individuals (veiled Arab women, veiled ethnic Danish Moslem women and Danish controls); and (ii) through food intake analysis to estimate the oral intake of vitamin D necessary to keep a normal vitamin D status in sunlight-deprived individuals. DESIGN: Cross-sectional study amongst randomly selected Moslem women of Arab origin living in Denmark. Age-matched Danish women were included as controls. To control for racial differences, a group of veiled ethnic Danish Moslem women (all Caucasians) was included. SETTING: Primary Health Care Centre, City Vest and Department of Endocrinology and Metabolism C, University Hospital of Aarhus, Aarhus Amtssygehus, Aarhus, Denmark. SUBJECTS: Sixty-nine Arab women (60 veiled, nine non-veiled) and 44 age matched Danish controls were randomly selected amongst patients contacting the primary health care centre for reasons other than vitamin D deficiency. Ten ethnic Danish Moslem women were included through a direct contact with their community. MAIN OUTCOME MEASURES: Serum levels of 25-hydroxyvitamin D were used as estimates of vitamin D status. Intact parathyroid hormone (PTH) was used to control for secondary hyperparathyroidism. Alkaline phosphatase and bone-specific alkaline phosphatase were used as markers for osteomalacic bone involvement. Oral intake of vitamin D and calcium were estimated through a historical food intake interview performed by a trained clinical dietician. RESULTS: Veiled Arab women displayed extremely low values of 25-hydroxyvitamin D: 7.1 +/- 1.1 nmol L-1, compared with 17.5 +/- 2. 3 (P < 0.002) in ethnic Danish Moslems and 47.1 +/- 4.6 (P < 10-17) in Danish controls. PTH was increased amongst veiled Arab women: 15. 6 +/- 1.8 pmol L-1, compared with 5.7 +/- 1.4 in ethnic Danish Moslems and 2.7 +/ 0.3 (P < 10-6) in Danish controls. The vitamin D intake (including food supplementation) was very low amongst Arab women: 1.04 microg day-1, compared with 13.53 amongst ethnic Danish Moslems and 7.49 amongst Danish controls (P < 0.0005). CONCLUSIONS: Severe vitamin D deficiency is prevalent amongst sunlight deprived individuals living in Denmark. In veiled Arab women, vitamin D deficiency is the result of a combination of limitations in sunlight exposure and a low oral intake of vitamin D. The oral intake of vitamin D amongst veiled ethnic Danish Moslems was, however, very high, at 13.53 microgram (approximately 600 IU), but they were still vitamin D-deficient. Our results suggest that the daily oral intake of vitamin D in sunlight-deprived individuals should exceed 600 IU; most probably it should be 1000 IU day-1 to secure a normal level of 25 hydroxyvitamin D. This finding is in contrast with the commonly used RDA (recommended daily allowance) for adults in Europe: 200 IU day-1. PMID- 10692091 TI - Calcaneal ultrasound measurements are determined by age and physical activity. Studies in two Swedish random population samples. AB - AIM: To present reference values and correlations with body composition, blood variables and lifestyle factors. SUBJECTS: Two random population samples from Goteborg, Sweden, one comprising 184 men and 455 women aged 25-64 years (MONICA) and the other 860 women aged 55-82 years (BEDA) were studied. METHODS: Calcaneal ultrasound measurement (LUNAR Achilles) and bioimpedance were measured. Smoking habits, coffee consumption, physical activity, psychological stress, education and marital status, as well as blood lipids, blood pressure, and fractures were studied. RESULTS: Broadband ultrasound attenuation and stiffness were higher in men than in women (P < 0. 001), but speed of sound did not differ between sexes. Speed of sound, broadband ultrasound attenuation and stiffness decreased with age (P < 0.001). In both sexes speed of sound, broadband ultrasound attenuation and stiffness correlated positively to body size variables, and negatively with smoking in women after adjustment for age. Speed of sound, broadband ultrasound attenuation and stiffness were positively related to physical activity in both sexes, and these relationships were the only ones that remained in multivariate analyses in addition to age (negative). Osteoporotic fractures increased with age. Speed of sound, broadband ultrasound attenuation and stiffness were lower amongst women with osteoporotic fractures. CONCLUSION: Speed of sound, broadband ultrasound attenuation and stiffness decreased with age and increased with physical activity, but body weight and height were not correlated in multivariate analyses. Osteoporotic fractures increased with age and were associated with lower calcaneal ultrasound values. PMID- 10692092 TI - Increased plasma concentrations of TGF-beta1 after hormone replacement therapy. AB - OBJECTIVES AND DESIGN: Hormone replacement therapy (HRT) in postmenopausal women may reduce the cardiovascular risk. A dominant protective role of transforming growth factor beta (TGF-beta1) on coronary arteries has been proposed. Lp(a) lipoprotein may block the activation of latent TGF-beta1. Given this background, we examined the effects of HRT on TGF-beta1 and Lp(a) lipoprotein in 99 postmenopausal women. The women had angiographically documented coronary heart disease (CHD) and were randomized to either sequential transdermal 17beta oestradiol for 14 weeks and then medroxyprogesterone (MPA) for 14 days (HRT) or to a control group (C). RESULTS: Serum levels of TGF-beta1 were increased in the HRT group compared with the C group after 3 months' treatment and this effect was sustained after 12 months. There was a significant reduction in Lp(a) lipoprotein serum levels after 3 months' treatment in the HRT group compared with the C group. However, after 12 months, no significant difference in changes in Lp(a) lipoprotein serum levels was detected between the two groups. CONCLUSION: The novel observation that transdermal 17beta-oestradiol in postmenopausal women increases levels of TGF-beta1 and lowers the concentration of Lp(a) lipoprotein suggests yet another possible mechanism for the cardioprotective effect of HRT. Whereas combination therapy of oestradiol and MPA preserves the beneficial effect on TGF-beta1, it reduces the unopposed oestradiol effects on Lp(a) lipoprotein. PMID- 10692093 TI - Plasma total homocysteine levels in subjects with hyperinsulinemia. AB - OBJECTIVES: Hyperhomocysteinemia as well as insulin resistance are considered to be risk factors for the development of coronary artery disease. This study was aimed at determining whether any relationship between plasma insulin and glucose levels and total plasma homocysteine (tHcy) concentrations exists in a population based survey performed 10 years apart. DESIGN AND SETTING: A cross-sectional study was undertaken during the years 1986-87 to examine risk factors for diabetes and for coronary artery disease (CAD) in the Jewish population of Jerusalem. Ten years later two groups of individuals were invited for re examination. SUBJECTS: Two groups of individuals were examined: the first one consisted of nondiabetic subjects (n = 86), who had hyperinsulinemia 10 years previously (at the first visit), the second group consisted of normoinsulinemic nondiabetic individuals (n = 265) who had initially normal glucose and insulin levels. MAIN OUTCOME MEASURES: Metabolic, biochemical and anthropomorphic features were determined. Fasting and post load glucose, as well as insulin concentrations on fasting and 2 h post glucose load were measured at the first and second visits. Plasma tHcy and folic acid were determined only at the second visit. RESULTS: The results demonstrated a significant negative correlation between plasma tHcy levels and insulin levels at the second visit. No difference was found in folic acid levels between these two groups. CONCLUSIONS: In general, hyperinsulinemia and hyperhomocysteinemia are both related to an increased incidence of CAD. In our population most of the subjects examined had tHcy levels within the normal range and only a few demonstrated very high levels. However, negative association between insulin levels and tHcy concentrations was found. Possible explanations for this finding are discussed. PMID- 10692094 TI - Coronary vasospasm associated with uncontrolled carcinoid tumour. AB - We report on a 74-year-old carcinoid patient who, following acute myocardial infarction (MI) and percutaneous transluminal coronary angioplasty, suffered recurrent episodes of chest pain and ST-segment elevation on ECG. This was accompanied by elevation of urinary 5-hydroxy-indole acetic acid. A review of the patient's file revealed that during the 3 weeks prior to the MI, she had been treated inadvertently with a fivefold lower dosage of octreotide. Following the correction of octreotide dosage, episodes of chest pain resolved immediately. We therefore suggest that this patient suffered from recurrent coronary vasospasm due to uncontrolled carcinoid tumour. PMID- 10692095 TI - In this issue PMID- 10692096 TI - Antibodies to human papillomavirus type 5 are generated in epidermal repair processes. AB - We reported previously that patients with psoriasis harbored at a very high frequency DNA sequences of the oncogenic human papillomavirus type 5 (HPV5) associated with epidermodysplasia verruciformis. Moreover anti-HPV5 antibodies were detected in 25% of the cases. Our aim was to find out whether keratinocyte hyperproliferation and/or autoimmunity could be responsible for HPV5 expression in psoriasis. We found that epidermal repair in patients with extensive second degree burns (n = 19) is frequently associated with the generation of anti-HPV5 antibodies. In patients with autoimmune bullous diseases (n = 118), a condition in which keratinocyte proliferation is involved in repair mechanisms, the prevalence of anti-HPV5 antibodies (15%-25%) was similar to that reported in psoriasis and significantly higher than that (5%) observed in individuals with no known history of human papillomavirus infection (n = 119). A high detection rate (57.9%) of HPV5 DNA was observed in patients with bullous diseases. Anti-HPV5 antibodies were found in patients with autoimmune connective tissue disorders with cutaneous involvement (n = 40) as frequently as in patients with bullous diseases. HPV5 DNA was detected in one of the 10 patients studied. In contrast, the prevalence of anti-HPV5 antibodies in patients with autoimmune neurological disorders (n = 47) and in patients with common warts (n = 28) or invasive carcinomas of the skin (n = 40) was as low as in the general population. It is worth stressing that a similar prevalence of antibodies against HPV1 was found in all groups studied. Our data strongly suggest that extensive keratinocyte proliferation is a major factor for the generation of anti-HPV5 antibodies and that autoimmunity may contribute to this phenomenon. It remains to be determined whether HPV5 and other human papillomavirus genotypes associated with epidermodysplasia verruciformis contribute to the hyperproliferation of keratinocytes occurring in epidermal repair and in psoriasis. PMID- 10692097 TI - Signaling of mechanical stretch in human keratinocytes via MAP kinases. AB - Cells within human skin are permanently exposed to mechanical stretching. Here we present evidence that alterations in cell shape trigger biochemical signaling via MAP kinases in human keratinocytes. In an in vitro attempt we demonstrate a fast but transient activation of extracellular signal-regulated kinases 1/2 in response to cell stretch. This activation is reversed by preincubation with functional blocking antibodies directed towards beta1-integrins. As a second member of MAP kinases, stress-activated protein kinase/c-JUN NH2-terminal kinase was activated in a slower fashion, peaking at 1 h after the initial stimulus. The delay in signal transmission suggests that extracellular signal-regulated kinases 1/2 and stress-activated protein kinase/c-JUN NH2-terminal kinase do not share the same signaling pathway. p38 was not activated by cell stretching. The contribution of cytoskeletal elements in signal perception and transduction was evaluated by selective disruption of either actin filaments, microtubules, or keratin filaments but showed no clear effect on stretch-induced activation of extracellular signal-regulated kinases 1/2 and stress-activated protein kinase/c JUN NH2-terminal kinase. In conclusion we found evidence of a cell-shape dependent activation of MAP kinases in human keratinocytes disclosing beta1 integrins as putative mechano-transducers. It is likely that alterations of skin mechanics in vivo underlying pathogenic processes like wound formation and healing trigger physiologic responses via the MAP kinase pathway. PMID- 10692098 TI - Adhesive properties of human basal epidermal cells: an analysis of keratinocyte stem cells, transit amplifying cells, and postmitotic differentiating cells. AB - The basal layer of human epidermis is a heterogeneous population of proliferative and differentiating cells that can be divided into at least three functionally discrete compartments: keratinocyte stem cells, transit amplifying cells, and postmitotic differentiating cells. Basal cells adhere to the underlying basement membrane via integrins, and although decreased adhesion is a key event in epidermal differentiation, the specific role of particular integrins is poorly understood. We report here on the comparative expression and function of the beta1 versus alpha6beta4 integrins in keratinocyte stem cells, transit amplifying cells, and postmitotic differentiating cells of neonatal human foreskin epidermis. Adhesion assays demonstrate that both keratinocyte stem cells and transit amplifying cells comprise rapidly adhering cells that exhibit high levels of functional beta1 and alpha6beta4 integrins. Interestingly, a proportion of basal cells that have begun to differentiate in vivo within the basal layer as determined by their expression of the differentiation-specific markers K10 and involucrin also retain high levels of activated beta1 integrin, but downregulate alpha6beta4 expression selectively (termed alpha6dimbeta1bri). These cells also retain their adhesive capacity, indicating that induction of differentiation in vivo does not correlate with decreased beta1 integrin expression or function. We have previously reported on the use of alpha6 integrin in conjunction with a proliferation associated marker (10G7 ag) to separate keratinocyte stem cells (phenotype alpha6bri10G7dim) from other basal cells (Li et al. Proc Natl Acad Sci 95:3902-3907 1998). A comparison of the long-term proliferative potential of beta1bri10G7dim cells with alpha6bri10G7dim showed that selection of alpha6bri10G7dim allows the isolation of a purer fraction of keratinocyte stem cells. PMID- 10692099 TI - Kinesin and kinectin can associate with the melanosomal surface and form a link with microtubules in normal human melanocytes. AB - Microtubuli play an important role in the organization of organelles and membrane traffic. They are present in melanocytic dendrites through which melanosomes are transported towards keratinocytes. Besides the actin-based motility systems, microtubuli-associated motor proteins also play a critical role in melanosome movement, as has recently been confirmed in mouse melanocytes. We investigated the in vitro expression of two forms of human conventional kinesin and its receptor kinectin in normal human epidermal melanocytes, keratinocytes, and dermal fibroblasts by reverse transcription polymerase chain reaction and northern blot analysis. In an attempt to gain insight into the subcellular distribution of kinesin and kinectin in melanocytes, double immunofluorescent staining and immunogold electron microscopy were performed. In all studied skin cells ubiquitous and neuronal kinesin are expressed, as well as the kinectin receptor. Immunofluorescent staining shows distinct but partially overlapping distributions for kinesin heavy chain and melanosomes, suggesting that kinesin is associated with some but not all of the melanosomes. Similar observations for kinectin indicate that this receptor can colocalize with melanosomes, which was confirmed by immunoelectron microscopy. The latter technique allowed us to demonstrate a close association between kinesin heavy chain, microtubuli, and melanosomes. The combined data from reverse transcription polymerase chain reaction, northern blot analysis, double immunofluorescent staining, and immunogold electron microscopy suggest that kinesins and kinectin have an important role in microtubuli-based melanosome transport in human melanocytes. PMID- 10692100 TI - Pro-opiomelanocortin-related peptides, prohormone convertases 1 and 2 and the regulatory peptide 7B2 are present in melanosomes of human melanocytes. AB - Recently, it has been shown that alpha-melanocyte stimulating hormone can directly activate tyrosinase by removing the allosteric regulator 6(R)-L-erythro 5,6,7,8 tetrahydrobiopterin resulting in a stable alpha-melanocyte stimulating hormone/6(R)-L-erythro 5,6,7,8 tetrahydrobiopterin complex. As melanin production occurs in the melanosome, a specific organelle of the melanocyte, it seemed important to investigate whether these organelles themselves actually produce pro opiomelanocortin-related peptides in their acidic environment. The presence of alpha-melanocyte stimulating hormone and adrenocorticotropin in the epidermis and melanocytes has been shown by several investigators. In order to follow possible pro-opiomelanocortin processing in the melanosome, human melanocytes were established in MCDB 153 medium and utilized for immunohistochemistry, immunogold electron microscopy, and western blotting. For this purpose antibodies against alpha-melanocyte stimulating hormone, adrenocorticotropin, prohormone convertases 1 and 2 (PC1 and PC2) and the PC2 regulatory protein 7B2 were used. Our results demonstrated the presence of the entire system for pro-opiomelanocortin processing in the melanosome. Considering the pH optima of these convertases, the results are in agreement with an autocrine intramelanosomal production of pro- opiomelanocortin-related peptides and an autocrine production and recycling of the cofactor 6(R)-L- erythro 5,6,7,8 tetrahydrobiopterin in melanocytes. Based on these novel observations, we would like to propose that the pigmentation process may not necessarily involve a melanocortin-1 receptor-mediated mechanism. PMID- 10692101 TI - Kinesin participates in melanosomal movement along melanocyte dendrites. AB - Movement of melanosomes along melanocyte dendrites is necessary for the transfer of melanin pigment from melanocytes to basal and suprabasal keratinocytes, an event critical to epidermal photoprotection and maintenance of normal skin color. Recent murine data suggest that in melanocyte dendrites the microtubule associated melanosome movement is bidirectional and that actin-associated myosin V secures the peripheral melanosomes, preparing them to be transferred to surrounding keratinocytes. We now report that human melanocytes express high levels of kinesin, a molecule that participates in microtubule-associated transport of organelles in other cell types, and that ultrastructurally kinesin molecules are closely associated with melanosomes. To determine whether kinesin participates in melanosomal transport, cultured melanocytes were treated with sense or antisense oligonucleotides complementary to kinesin heavy chain sequences. Antisense oligonucleotides decreased kinesin protein levels and inhibited the bidirectional movement of the melanosomes, promoting their backward movement. Furthermore, guinea pigs were exposed to ultraviolet B irradiation, known to enhance transport of melanosomes from melanocytes to epidermal keratinocytes, and then were treated with kinesin sense or antisense oligonucleotides. The areas that were treated with kinesin antisense oligonucleotides showed significantly less pigmentation clinically and histologically than control (sense) oligonucleotide-treated areas. As observed ultrastructurally, in antisense-treated areas melanosomes remained in melanocyte dendrites but over several days were not transferred to the surrounding keratinocytes. Our study supports a major role for kinesin in microtubule associated anterograde melanosomal transport in human melanocyte dendrites. PMID- 10692102 TI - Normal growth and differentiation in a spontaneously immortalized near-diploid human keratinocyte cell line, NIKS. AB - We report the isolation and characterization of a spontaneously immortalized human keratinocyte cell line, NIKS. The cell line is not tumorigenic in athymic nude mice and maintains cell-type-specific requirements for growth in vitro. NIKS cells express steady-state levels of transforming growth factor-alpha, transforming growth factor-beta1, epidermal growth factor receptor, c-myc, and keratin 14 mRNAs comparable with the parental BC-1-Ep keratinocyte strain. BC-1 Ep and NIKS keratinocytes produce similar levels of cornified envelopes and nucleosomal fragmentation in response to loss of substrata attachment. DNA fingerprinting results confirm that the NIKS cells originated from the parental BC-1-Ep keratinocytes. NIKS cells contain 47 chromosomes due to an extra isochromosome of the long arm of chromosome 8, and the near-diploid karyotype appears to be stable with repeated passage. A fully stratified squamous epithelium is formed by the NIKS keratinocytes in organotypic culture. Ultrastructural analysis of both the parental and immortalized keratinocytes reveals abundant desmosomes, hemidesmosomes, and the production of a basal lamina. Our findings with the NIKS cells support the observation that spontaneous immortalization is not linked to alterations in squamous differentiation or the ability to undergo apoptosis. The NIKS human keratinocyte cell line is an important new tool for the study of growth and differentiation in stratified squamous epithelia. PMID- 10692103 TI - Dual-mode regulation of hair growth cycle by two Fgf-5 gene products. AB - As the result of alternative mRNA splicing, Fgf-5, the gene encoding fibroblast growth factor-5, translates to both long and short forms of the protein, respectively, designated fibroblast growth factor-5 and fibroblast growth factor 5S. We previously showed that localization of fibroblast growth factor-5 and the level of fibroblast growth factor-5S in murine skin are hair-cycle dependent. In this study, we examined the effect of fibroblast growth factor-5 and fibroblast growth factor-5S on the hair growth cycle in mice. Once the anagen phase of the hair growth cycle was induced in the dorsal skin by depilation during telogen, and effects of subcutaneous injection of fibroblast growth factor-5 and fibroblast growth factor-5S into the affected region were analyzed. We found that fibroblast growth factor-5 inhibited hair growth during anagen and promoted the transition from anagen to catagen. Interestingly, whereas fibroblast growth factor-5S alone exerted no effect on hair growth, it significantly inhibited the catagen-promoting activity of fibroblast growth factor-5 when the two proteins were injected simultaneously. Because neither fibroblast growth factor-5 nor fibroblast growth factor-5S affected skin thickness, it is postulated that changes in skin thickness during hair cycle are separately regulated by factors other than those regulating hair and follicle growth. The present results, together with our earlier findings that fibroblast growth factor-5-producing cells gather around dermal papillae during catagen, whereas fibroblast growth factor-5S is abundantly expressed in the hair follicles only during the latter half of anagen, suggests that the mouse hair growth cycle is regulated by the two Fgf-5 gene products acting in concert: fibroblast growth factor-5 induces catagen, whereas fibroblast growth factor-5S antagonizes this activity during anagen. PMID- 10692104 TI - Characterization of a 300 kbp region of human DNA containing the type II hair keratin gene domain. AB - Screening of an arrayed human genomic P1 artificial chromosome DNA library by means of the polymerase chain reaction with a specific primer pair from the human type II hair keratin hHb5 yielded two P1 artificial chromosome clones covering approximately 300 kb of genomic DNA. The contig contained six type II hair keratin genes, hHb1-hHb6, and four keratin pseudogenes psihHbA-psihHbD. This hair keratin gene domain was flanked by type II epithelial keratins K6b/K6hf and K7, respectively. The keratin genes/pseudogene are 5-14 kbp in size with intergenic distances of 5-19 kbp of DNA and do not exhibit a single direction of transcription. With one exception, type II hair keratin genes are organized into nine exons and eight introns, with strictly conserved exon-intron boundaries. The functional hair keratin genes are grouped into two distinct subclusters near the extremities of the hair keratin gene domain. One subcluster encodes the highly related hair keratins hHb1, hHb3, and hHb6; The second cluster encodes the structurally less related hair keratins hHb2, hHb4, and hHb5. Reverse transcription-polymerase chain reaction shows that all hair keratin genes are expressed in the hair follicle. Pseudogene psihHbD is also transcriptionally expressed, albeit with alterations in splicing and frameshift mutations, leading to premature stop codons in the splice forms analyzed. Evolutionary tree analysis revealed a divergence of the type II hair keratin genes from the epithelial keratins, followed by their segregation into the members of the two subclusters over time. We assume that the approximately 200 kbp DNA domain contains the entire complement of human type II hair keratin genes. PMID- 10692105 TI - New function for NF1 tumor suppressor. AB - The expression and subcellular localization of neurofibromatosis type 1 tumor suppressor was studied in keratinocytes induced to differentiate by increased Ca2+ concentration of the culture medium. Differentiating keratinocytes became intensely immunoreactive for neurofibromatosis type 1 protein, which was apparently associated with cellular fibrils. Double immunolabeling with antibodies to cytokeratin 14 and neurofibromatosis type 1 protein suggested an association of intermediate type cytoskeleton and neurofibromatosis type 1 protein. The presence of neurofibromatosis type 1 protein in cell preparations treated with cytoskeletal buffer indicated a high affinity interaction between intermediate filaments and neurofibromatosis type 1 protein. Further studies utilizing double immunolabelings revealed that the intense neurofibromatosis type 1 tumor suppressor signal on intermediate filaments was temporally limited to the period in keratinocyte differentiation in which the formation of desmosomes takes place. Keratinocytes were also cultured from nine patients with type 1 neurofibromatosis and were studied with respect to cell morphology, and association of neurofibromatosis type 1 protein with intermediate cytoskeleton. The results showed that keratinocytes cultured from patients with neurofibromatosis type 1 displayed a highly variable cell size and morphology compared to controls. The latter findings represent predicted alterations in a situation where cytoskeletal organization is disturbed. Furthermore, differentiating neurofibromatosis type 1 keratinocytes were characterized by a reduced number of cytokeratin bundles that were decorated neurofibromatosis type 1 protein. The results of this study suggest that neurofibromatosis type 1 tumor suppressor exerts its effects in part by controlling organization of cytoskeleton during the formation of cellular contacts. PMID- 10692106 TI - Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. AB - Damage to human skin due to ultraviolet light from the sun (photoaging) and damage occurring as a consequence of the passage of time (chronologic or natural aging) are considered to be distinct entities. Photoaging is caused in part by damage to skin connective tissue by increased elaboration of collagen-degrading matrix metalloproteinases, and by reduced collagen synthesis. As matrix metalloproteinase levels are known to rise in fibroblasts as a function of age, and as oxidant stress is believed to underlie changes associated with both photoaging and natural aging, we determined whether natural skin aging, like photoaging, gives rise to increased matrix metalloproteinases and reduced collagen synthesis. In addition, we determined whether topical vitamin A (retinol) could stimulate new collagen deposition in sun-protected aged skin, as it does in photoaged skin. Sun-protected skin samples were obtained from 72 individuals in four age groups: 18-29 y, 30-59 y, 60-79 y, and 80+ y. Histologic and cellular markers of connective tissue abnormalities were significantly elevated in the 60-79 y and 80+ y groups, compared with the two younger age groups. Increased matrix metalloproteinase levels and decreased collagen synthesis/expression were associated with this connective tissue damage. In a separate group of 53 individuals (80+ y of age), topical application of 1% vitamin A for 7 d increased fibroblast growth and collagen synthesis, and concomitantly reduced the levels of matrix-degrading matrix metalloproteinases. Our findings indicate that naturally aged, sun-protected skin and photoaged skin share important molecular features including connective tissue damage, elevated matrix metalloproteinase levels, and reduced collagen production. In addition, vitamin A treatment reduces matrix metalloproteinase expression and stimulates collagen synthesis in naturally aged, sun-protected skin, as it does in photoaged skin. PMID- 10692107 TI - Keratin 4 upregulation by retinoic acid in vivo: a sensitive marker for retinoid bioactivity in human epidermis. AB - Retinoids affect keratinocyte differentiation and modulate the expression of many epidermal proteins, among them cellular retinoic acid-binding protein II and the family of cytokeratins. The upregulation of the former protein is a well-known phenomenon, whereas the retinoid-induced regulation of epidermal keratin expression is more complex and only partially understood. We studied the effect of topical retinoids on the expression in healthy skin of cellular retinoic acid binding protein II, tazarotene-induced genes 1 and 2, several epidermal keratins (K1, K2e, and K10), and two mucous keratins (K4 and K13) known to appear in epidermis under certain abnormal conditions. Reverse transcription-polymerase chain reaction experiments showed that the K4 expression was the one most overtly induced by 2 wk of open treatment with 0.05% of retinoic acid and tazarotene. Using real-time quantitative polymerase chain reaction (TaqMan) and normalization of the mRNA values to beta-actin, the increase in K4 was found to be 100-1000 fold. In comparison, the expression of K13 and cellular retinoic acid-binding protein II was increased 10-50-fold, the K1 and K10 mRNA levels remained unchanged, and the K2e level decreased by a factor of 100-1000. In parallel biopsies, immunohistochemistry showed no change in K1, K2e, or K10 staining, but a strong de novo appearance of K4 in the granular layer after retinoid treatment. In a separate study, occlusive application of 0.025% retinoic acid in four healthy subjects produced a maximal K4 mRNA signal after 48 h and strong K4 staining after 80 h. Finally, a dose-response study showed that the de novo appearance of K4 can be utilized as a sensitive test for retinoid bioactivity in epidermis in vivo. PMID- 10692108 TI - Suppression of vitamin D receptor and induction of retinoid X receptor alpha expression during squamous differentiation of cultured keratinocytes. AB - To gain more insight in the role of the vitamin D system in epidermal differentiation, we studied the expression of the vitamin D receptor and its heterodimeric partner retinoid X receptor alpha in cultured normal human keratinocytes during squamous differentiation, as triggered by different approaches. Northern and western blot analysis allowed us to investigate mRNA and protein levels of these nuclear receptors and of markers for growth control (c myc, cyclin D1, p21WAF1) and differentiation (keratinocyte transglutaminase, small proline rich proteins). Growing cells to postconfluence was a potent stimulus for growth arrest and differentiation with concomitant suppression of vitamin D receptor and induction of retinoid X receptor alpha, at both the mRNA and the protein level. These changes could be prevented by concomitant treatment with epidermal growth factor or keratinocyte growth factor. Subjecting the cells to a calcium switch leading to stratification and differentiation lowered vitamin D receptor protein levels without affecting vitamin D receptor mRNA and induced both retinoid X receptor alpha mRNA and protein. Interferon-gamma and the phorbolester 12-O-tetradecanoyl phorbol 13-acetate, two well-known inducers of keratinocyte differentiation, both inhibited vitamin D receptor expression but only interferon-gamma induced retinoid X receptor alpha. The decreased vitamin D receptor expression was accompanied by reduced vitamin D responsiveness (as assessed by 24-hydroxylase mRNA induction) in postconfluent, high calcium, and 12 O-tetradecanoyl phorbol 13-acetate treated keratinocytes but not with interferon gamma treatment. Taken together, our results associate vitamin D receptor expression with undifferentiated, proliferating keratinocytes, whereas retinoid X receptor alpha expression appears to be related to the differentiated phenotype. Therefore, proliferating and differentiating keratinocytes may be differentially targeted by active vitamin D metabolites. PMID- 10692109 TI - Protective role of copper, zinc superoxide dismutase against UVB-induced injury of the human keratinocyte cell line HaCaT. AB - On the basis of our recent observation that copper, zinc-superoxide dismutase and manganese-superoxide dismutase change differently following a single exposure to ultraviolet-B irradiation in the human keratinocyte cell line HaCaT, we have examined the possible role of endogenous copper,zinc-superoxide dismutase or manganese-superoxide dismutase against ultraviolet-B-induced reactive-oxygen- species-mediated keratinocyte injury in vitro. To evaluate the individual defensive roles of copper, zinc-superoxide dismutase and manganese-super-oxide dismutase, we treated HaCaT cells with diethyldithiocarbamate, a chelating agent of ionic copper that inactivates copper,zinc-superoxide dismutase activities, tumor necrosis factor alpha, which enhances manganese-superoxide dismutase levels, or transforming growth factor beta1, which inhibits manganese-superoxide dismutase levels. After the treatment with each reagent, HaCaT cells in the three different conditions were exposed to a single dose of ultraviolet-B irradiation. We assessed ultraviolet-B-induced cytotoxicity by measuring both lactate dehydrogenase leakage and cell viability using trypan blue dye exclusion assay. The lactate dehydrogenase leakage in the supernatant from damaged HaCaT cells whose copper,zinc-superoxide dismutase levels were inactivated by diethyldithiocarbamate was significantly increased and the cell viability was significantly decreased in comparison with untreated groups at 8 and 24 h after ultraviolet-B irradiation. On the other hand, the lactate dehydrogenase release and cell viability for HaCaT cells whose manganese-superoxide dismutase levels were enhanced by tumor necrosis factor alpha or inhibited by transforming growth factor beta1 showed no significant difference from untreated groups. Furthermore, increased production of intracellular peroxides in HaCaT cells treated with diethyldithiocarbamate was observed by flow cytometric analysis at 8 h after ultraviolet-B irradiation. These results suggest that copper,zinc-superoxide dismutase may play a primary protective role against ultraviolet-B-induced injury of the human keratinocyte cell line HaCaT. PMID- 10692110 TI - Ultraviolet B-induced suppression of immune responses in interleukin-4-/- mice: relationship to dermal mast cells. AB - Ultraviolet B radiation is immunosuppressive by multiple mechanisms. In interleukin-4-/- mice, ultraviolet B radiation was not able to suppress delayed type hypersensitivity or contact hypersensitivity responses when the sensitizing antigen was applied to nonirradiated sites. In contrast, ultraviolet B significantly suppressed contact hypersensitivity responses to haptens applied to irradiated sites in interleukin-4-/- mice. In mast cell depleted Wf/Wf mice, ultraviolet B radiation also significantly suppressed contact hypersensitivity responses to sensitizing antigens applied to irradiated but not to unirradiated sites. In both interleukin-4-/- mice and Wf/Wf mice, the mast cell product, histamine, was immunosuppressive implicating mast cells as the dysfunctional cell in interleukin-4-/- mice. The prevalence of dermal mast cells was similar in wild type and interleukin-4-/- mice. Dermal mast cells of interleukin-4-/- mice, however, express very low levels of c-kit and did not significantly degranulate in response to ultraviolet B. Ultraviolet radiation induced significant and similar levels of serum interleukin-10 in wild-type and interleukin-4-/- mice. We conclude that interleukin-4 indirectly affects ultraviolet B suppression of contact hypersensitivity and delayed-type hypersensitivity responses to sensitizing antigens applied at sites other than those irradiated by providing a critical differentiative signal for dermal mast cells. This study further emphasizes the central role of mast cells in the initial processes by which ultraviolet B radiation is immunomodulatory for immune responses to sensitizing antigens applied to nonirradiated sites. PMID- 10692111 TI - p53-dependent apoptosis in melanoma cells after treatment with camptothecin. AB - Cutaneous malignant melanoma is a life-threatening cancer with poor prognosis due to a high metastasis potential. The main obstacle in treatment of metastatic melanoma is the resistance to chemotherapy. Recent studies indicated that apoptosis is a common mechanism of action for various cytotoxic agents. As p53 plays an important part in apoptosis, we investigated the role of p53 in chemosensitivity of melanoma cells. Previously, we found that melanoma cell lines containing wild-type p53 have significantly higher response rates to chemotherapy than cell lines with a mutant p53 gene. To confirm the role of p53 in melanoma chemosensitivity further, we transfected an expression vector, pED1, which carries a mutant p53 gene, into a wild-type p53 melanoma cell line, MMAN. We examined the effect of mutant p53 on camptothecin-induced apoptosis and the expression of genes which are known to be involved in apoptosis or drug resistance, such as bcl-2, bax, bak, p21waf1, and P-glycoprotein. Our results indicate that overexpression of the mutant p53 increased the growth rate of MMAN cells, reduced the sensitivity to camptothecin, and lowered drug-induced apoptosis by 2-3-fold. Flow cytometry indicated that the camptothecin-induced apoptosis is not associated with G1 arrest. Furthermore, camptothecin treatment reduced bcl-2 and P-glycoprotein expression in wild-type p53 MMAN cells, but not cells overexpressing mutant p53. These results demonstrate that p53 mutational status is a determinant of melanoma chemosensitivity. p53 may downregulate bcl-2 and P-glycoprotein to induce apoptosis in melanoma cells after chemotherapy. PMID- 10692112 TI - alpha-melanocyte-stimulating hormone modulates nitric oxide production in melanocytes. AB - We have previously observed that melanocytes produce nitric oxide in response to ultraviolet radiation and lipopolysaccharide and in this study have examined how these responses are affected by alpha-melanocyte-stimulating hormone. Nitric oxide production by cultured cells was measured electrochemically in real time using an ISO-nitric oxide sensor probe. B16 mouse melanoma cells released nitric oxide in response to lipopolysaccharide and the effects were enhanced in cells that had been grown in the presence of 10-11-10-9 M alpha-melanocyte-stimulating hormone prior to stimulation. At concentrations in excess of 10-9 M alpha melanocyte-stimulating hormone decreased nitric oxide production. Preincubation with lipopolysaccharide, a well-known inducer of inducible nitric oxide synthase, also increased nitric oxide production but this response was reduced by alpha melanocyte-stimulating hormone. alpha-Melanocyte-stimulating hormone also increased the levels of nitric oxide produced in response to ultraviolet radiation (20-100 mJ per cm2) in B16 cells. The same effect was seen in human melanocytes and as this was inhibited by aminoguanidine would appear to involve an induction of inducible nitric oxide synthase. Reverse transcription-polymerase chain reaction showed that melanocytic cells express inducible nitric oxide synthase mRNA. Western blotting analysis and immunocytochemistry confirmed the presence of inducible nitric oxide synthase protein in B16 cells and FM55 human melanoma cells and that the levels were increased in response to alpha-melanocyte stimulating hormone. alpha-Melanocyte-stimulating hormone, however, decreased inducible nitric oxide synthase protein expression, which occurred in response to lipopolysaccharide. These results suggest that alpha-melanocyte-stimulating hormone regulates nitric oxide production in melanocytic cells by modulating the induction of inducible nitric oxide synthase. Additional experiments showed that nitric oxide increased melanin production by B16 cells and human melanocytes. This is in keeping with a melanogenic role for nitric oxide but whether its production by melanocytes in response to alpha-melanocyte-stimulating hormone is associated with such a role or whether it has some other significance relating to melanocyte differentiation or in mediating immunomodulatory actions of alpha melanocyte-stimulating hormone remains to be seen. PMID- 10692113 TI - beta-endorphin stimulates cytokeratin 16 expression and downregulates mu-opiate receptor expression in human epidermis. AB - It has been reported that opioid peptides modulate the differentiation of normal human keratinocytes and that mu-opiate receptors are expressed in human epidermis. The regulation of keratinocyte differentiation is particularly important in psoriasis, and one of the markers for hyperproliferative and differentiating skin diseases is cytokeratin 16. The finding that the endogenous mu-opiate receptor ligand beta-endorphin is increased in serum of patients with psoriasis indicates that the mu-opiate system may play an important role in the pathophysiology of the skin. In this study, we addressed the question whether there is a link between mu-opiate receptor regulation and cytokeratin 16 expression in normal and psoriatic skin. Firstly, we demonstrate that beta endorphin concentrations between 16 and 1000 nM significantly downregulate mu opiate receptor expression in epidermis of cultured human skin after 48 h. Secondly, we show that beta-endorphin regulates cytokeratin 16 expression in the epidermis of skin organ cultures exposed to 41-125 nM beta-endorphin for 48 h, leading to elevated cytokeratin 16 production. As expected, the expression of cytokeratin 16 was detected primarily in the suprabasal layer. The same pattern was observed in psoriatic lesional skin, i.e., mu-opiate receptor expression was significantly downregulated and cytokeratin 16 expression upregulated. These results suggest that the mu-opiate receptor system and its ligand beta-endorphin are involved in the pathogenesis of psoriasis, especially in terms of differentiation. PMID- 10692114 TI - The transepidermal oxygen flux from the environment is in balance with the capillary oxygen supply. AB - It has been known since the nineteenth century that oxygen is taken up by the human skin. With a newly developed sensor it became possible to examine the influence of the vascular supply on the oxygen flux into the skin, tcJ(O2). tcJ(O2) was measured optically by determining the oxygen partial pressure difference, DeltapO2 across a diffusion test membrane, which itself was brought into close contact to the skin surface. Under these conditions DeltapO2 is proportional to the tcJ(O2). The skin perfusion was varied by the application of a hyperemizing ointment on the abdomen of 12 volunteers and by suprasystolic occlusion at the thigh of 20 volunteers. The tcJ(O2) was measured at a temperature of 33 degrees C of the humid skin. It was compared with the skin perfusion monitored by laser Doppler flow, and the capillary oxygen supply measured by transcutaneous partial pressure of oxygen, tcpO2, at an electrode temperature of 37 degrees C. The transcutaneous O2 flux produced a distinct DeltapO2 of 81.8 +/- 8.2 Torr (abdomen) and 72.8 +/- 12.3 Torr (ankle). In hyperemic skin on the abdomen the O2 flux was reduced (DeltapO2 = 57.7 +/- 10.6 Torr). The tcpO2 increased from 8.7 +/- 10.7 to 35.1 +/- 16.9 Torr. During suprasystolic occlusion, DeltapO2 increased by 6.4 +/- 2.3 Torr, whereas laser Doppler flow and tcpO2 decreased significantly. These results indicate that the total oxygen supply of the epidermis and the upper dermis is guaranteed even if the perfusion varies. PMID- 10692115 TI - Regulation of cutaneous allergic reaction by odorant inhalation. AB - Olfactory stimuli modulate emotional conditions and the whole body immune system. Effects of odorant inhalation on cutaneous immune reaction were examined. Contact hypersensitivity to 2,4, 6-trinitrochlorobenzene was elicited in C57BL/6 mice. The reaction was suppressed at both the induction and elicitation phases by exposure to an odorant, citralva. Topical application of citralva or lyral/lilial did not affect the reaction. The suppressive effect of citralva was more potent than that of another odorant, lyral/lilial. Citralva decreased the number of epidermal Langerhans cells, whereas lyral/lilial had a weak effect. Citralva but not lyral/lilial induced plasma corticosterone. Glucocorticoid receptor antagonist abrogated the suppressive effect of citralva on contact hypersensitivity. Serum interleukin-12 was downregulated by exposure to citralva or lyral/lilial. These data demonstrate that olfactory stimuli regulate the cutaneous immune system. PMID- 10692116 TI - Oxysterols induce differentiation in human keratinocytes and increase Ap-1 dependent involucrin transcription. AB - Ligands and activators of the nuclear hormone receptor superfamily are important in the regulation of epidermal development and differentiation. Previously, we showed that naturally occurring fatty acids, as well as synthetic ligands for the peroxisome proliferator-activated receptor, induce keratinocyte differentiation in vitro. Here we asked whether oxysterols, another class of lipids formed de novo in the epidermis and that activate liver X-activated receptor, regulate keratinocyte differentiation. mRNA and protein levels of involucrin and transglutaminase 1, markers of differentiation, increased 2- to 3-fold in normal human keratinocytes incubated in the presence of 25- or 22R-hydroxycholesterol in low calcium. In high calcium, which alone induces differentiation, mRNA levels were further increased by oxysterols. Rates of cornified envelope formation, an indicator of terminal differentiation, also increased 2-fold with oxysterol treatment. In contrast, the rate of DNA synthesis was inhibited approximately 50% by oxysterols. Transcriptional regulation was assessed in keratinocytes transfected with either transglutaminase 1 or involucrin promoter-luciferase constructs. 22R-hydroxycholesterol increased transglutaminase 1 and involucrin promoter activity 2- to 3-fold. Either deletion of the -2452 bp to -1880 bp region of the involucrin promoter, or mutation of the AP-1 site within this region, abolished oxysterol responsiveness. Moreover, increased AP-1 DNA binding was observed in oxysterol-treated keratinocytes by gel shift analyses. Finally, we demonstrated the presence of liver X-activated receptor alpha and beta mRNAs, and showed that oxysterols stimulate a liver X-activated receptor response element transfected into keratinocytes. These data suggest that oxysterols induce keratinocyte differentiation, in part through increased AP-1-dependent transcription of the involucrin gene, an effect that may be mediated by liver X activated receptor. PMID- 10692117 TI - Stereoselective biosynthesis of hepoxilin B3 in human epidermis. AB - We previously reported that normal human epidermis forms 12-oxo-eicosatetraenoic acid and hepoxilin B3 as major eicosanoids and that hepoxilins and trioxilins are dramatically elevated in psoriatic lesions. We also observed that normal epidermis only synthesized one of the two possible 10-hydroxy- epimers of hepoxilin B3, suggesting its enzymatic origin. This study investigated the enzymatic pathways involved in the formation of hepoxilin B3 in human epidermis. Human epidermal fragments or cell fractions were incubated with [14C]-arachidonic acid or authentic 12(S)-hydroperoxyeicosatetraenoic acid. Products were analyzed by high-performance liquid chromatography, gas chromatography-mass spectrometry or a combination of both techniques. Esculetin and nordihydroguaiaretic acid inhibited formation of hepoxilin B3, 12-oxo-eicosatetraenoic acid, trioxilins, and 12-hydroxyeicosatetraenoic acid in a concentration-dependent manner. 12 Lipoxygenase activity was mainly located in the microsomal fraction (100,000 x g pellet) and 12-hydroxyeicosatetraenoic acid, hepoxilin B3, and 12-oxo eicosatetraenoic acid were formed. The hepoxilin B3-synthesizing activity was not observed in subcellular fractions incubated with authentic 12(S) hydroperoxyeicosatetraenoic acid, although it was located at least in the microsomal fraction when incubated with arachidonic acid. Similar results were obtained using preparations of recombinant platelet-type 12-lipoxygenase that yielded 12-oxo-eicosatetraenoic acid and hepoxilin B3 in addition to 12 hydroxyeicosatetraenoic acid, when incubated with arachidonic acid but not when incubated with 12-hydroperoxyeicosatetraenoic acid. Nevertheless, recombinant 12 lipoxygenase produced a lower ratio of 12-oxo-eicosatetraenoic acid and hepoxilin B3-12-hydroxyeicosatetraenoic acid than epidermis. Our results support the concept that 12-lipoxygenase catalyzes the formation of hepoxilin B3 and 12-oxo eicosatetraenoic acid. PMID- 10692118 TI - Langerhans cells migrate to local lymph nodes following cutaneous infection with an arbovirus. AB - Whereas there has been recent interest in interactions between dendritic cells and pathogenic viruses, the role of dendritic cells in the initiation of protective immunity to such organisms has not been elucidated. The aim of this study was to examine whether a resident dendritic cell population in the skin, Langerhans cells, respond to cutaneous viral infections which are effectively cleared by the immune system. We therefore characterized the ability of Langerhans cells to migrate to local draining lymph nodes following infection with the arthropod-borne viruses, West Nile virus or Semliki Forest virus. The data show that major histocompatibility complex class II+/NLDC145+/E-cadherin+ Langerhans cell numbers are increased in the draining lymph nodes of infected mice and this increase is accompanied by a concomitant decrease in the Langerhans cell density in the epidermis. Langerhans cell migration is associated with an accumulation of leukocytes in the lymph node, which is one of the earliest events in the initiation of an immune response. Both the migratory response and the draining lymph node leukocyte accumulation were abrogated if ultraviolet inactivated instead of live viruses were used, suggesting the activation and subsequent migration of Langerhans cells requires a live, replicating antigen. Our findings are likely to have wider implications for the development of epidermally delivered vaccines and suggest that mobilization of dendritic cells may be involved in the development of immune responses to arthropod-borne viruses. PMID- 10692119 TI - Formation of antigenic quinolone photoadducts on Langerhans cells initiates photoallergy to systemically administered quinolone in mice. AB - Quinolone antibacterial agents are well known to cause photoallergy as a side effect. Murine photoallergy to fluoroquinolones is a T cell-mediated immune response, evoked either by systemic fluoroquinolone and subsequent exposure of skin to ultraviolet A light or by subcutaneous injection of fluoroquinolone photomodified epidermal cells. In this photosensitivity, epidermal Langerhans cells may be photomodified initially with the drug and thus present photohaptenic moieties to sensitize and restimulate T cells. Although we have shown that Langerhans cells photocoupled in vitro with fluoroquinolones are capable of stimulating sensitized T cells, it remains unclear whether systemically given fluoroquinolone photomodifies Langerhans cells upon ultraviolet A irradiation of the skin and the Langerhans cells become photohapten-bearing, T cell-stimulatory cells. In a murine model of fleroxacin photoallergy induced by intraperitoneal injection of the drugs plus ultraviolet A irradiation of skin, we found that Langerhans cells as well as keratinocytes are photoderivatized with fleroxacin as demonstrated with a fluoroquinolone-specific monoclonal antibody. Langerhans-cell enriched epidermal cells prepared from mice treated with fleroxacin and ultraviolet A induced proliferation of sensitized T cells, indicating that photomodified Langerhans cells are functional. There was an optimal range of ultraviolet A dose to quantitatively and qualitatively form fleroxacin photomodified Langerhans cells, as excess ultraviolet A rather reduced the photoantigen-presenting capacity of Langerhans cells presumably because of drug phototoxicity. Our study suggests that Langerhans cells serve as photoantigen presenting cells in drug photoallergy. PMID- 10692120 TI - Skin CD4+ T cells produce interferon-gamma in vitro in response to streptococcal antigens in chronic plaque psoriasis. AB - Recently, we have demonstrated that group A streptococcal antigen reactive T cells are present in the skin lesions of chronic plaque psoriasis. To determine the cytokine profile (interferon-gamma, interleukin-4 and interleukin-10) of these T cells in response to streptococcal antigens, T cell lines were cultured from untreated lesional skin of 13 patients with chronic plaque psoriasis and 12 patients with other inflammatory skin diseases. T cell lines were incubated with or without a sonicated heat-killed mixture of group A streptococcal isolates for 18 h in the presence of a transport inhibitor, stained for surface CD4 or CD8 and intracellular cytokine expression, and analyzed by flow cytometry. Psoriatic T cell lines were grown from 10 of 13 patients and were predominately CD4+ (64% 85%) with 10%-32% CD8+ T cells. Variable numbers of CD4+ T cells produced interferon-gamma (0.8%-35%, median 13.9) in eight of 10 T cell lines (p < 0.02). In contrast, CD4+ T cells in five of 12 T cell lines obtained from disease controls did not produce or produced minimal interferon-gamma in response to group A streptococcal isolates; this was significantly different from the psoriatic T cell lines (p < 0.05). Small numbers of interleukin-10 positive (0.8% 1.3%) and interleukin-4 positive (2.1%-2.5%) CD4+ T cells induced by group A streptococcal isolates were also present in two out of five and three out of five psoriatic T cell lines, respectively. This was significantly less in each case than the numbers of CD4+/interferon-gamma+ T cells (p < 0.05). Cytokine-positive CD8+ T cells were rarely observed. These findings demonstrate that a subpopulation of CD4+ T cells in chronic plaque psoriasis skin lesions produces interferon-gamma in response to streptococcal antigens and may be relevant to the pathogenesis of psoriasis. PMID- 10692121 TI - CD40 ligation alters the cell cycle of differentiating keratinocytes. AB - CD40 is expressed in normal human keratinocytes, especially in the basal cell layer. We have recently reported that CD40 ligation strongly inhibits keratinocyte proliferation and induces their differentiation. In this study, the CD40 pathway that prevents keratinocyte growth was investigated. We first reported that interferon-gamma treatment potentiated the CD40-mediated inhibition of keratinocyte proliferation. CD40-CD40 ligand interactions, in the presence or absence of interferon-gamma, neither enhanced spontaneous keratinocyte apoptosis, nor did it enhance apoptosis induced by various agents. More importantly, we showed that CD40 signaling altered the keratinocyte cell cycle, as demonstrated by a decreasing number of cells in the G1 and S phases and an accumulation in G2/M phase of the cell cycle. Furthermore, western blot analysis of cell cycle regulatory proteins, showed a decrease in cyclin A and E expression in CD40 activated keratinocytes. Collectively, these results indicate that CD40 ligation inhibits keratinocyte renewal by a mechanism independent of cell apoptosis and that modulation of the keratinocyte cell cycle is an additional outcome of CD40 signaling. PMID- 10692122 TI - Psoriasis, its treatment, and cancer in a cohort of Finnish patients. AB - This study was designed to estimate the relative cancer risk of patients with moderate to severe psoriasis, with reference to different treatments. A cohort of 5687 hospitalized patients with psoriasis obtained from the Finnish Hospital Discharge Register in 1973-84 was linked with the records of the Finnish Cancer Registry. Standardized incidence ratios for cancer were calculated by dividing the observed number of cases by the expected cases, which were based on the national sex-specific and age-specific cancer incidence rates. By the end of 1995, 533 cancer cases were observed in the cohort. The overall cancer incidence was increased (standardized incidence ratio 1.3, 95% confidence interval 1.2 1.4). The estimated relative risks were highest for Hodgkin's disease (standardized incidence ratio 3.3, 95% confidence interval 1.4-6.4), squamous cell skin carcinoma (standardized incidence ratio 3.2, 95% confidence interval 2.3-4.4), non-Hodgkin's lymphoma (standardized incidence ratio 2.2, 95% confidence interval 1.4-3.4), and laryngeal cancer (standardized incidence ratio 2.9, 95% confidence interval 1.5-5.0). The role of prior oral antipsoriatic medications or phototherapy on the development of these cancers was assessed in a nested case-control study, for which 67 cases and 199 sex and age matched controls were selected from the psoriasis cohort. The relative risks were estimated using conditional logistic regression analysis. Oral 8-methoxy-psoralen plus ultraviolet-A radiation therapy and the use of retinoids were associated with an increased risk of squamous cell skin carcinoma (relative risk adjusted for the other treatment variables 6.5, 95% confidence interval 1.4-31, and 7.4, 95% confidence interval 1.4-40, respectively), whereas none of the treatments could be linked with the occurrence of non-Hodgkin's lymphoma. PMID- 10692123 TI - Mutation report: a novel partial deletion of exons 2-10 of the STS gene in recessive X-linked ichthyosis. AB - X-linked ichthyosis is an inherited disease due to steroid sulfatase deficiency. Onset is at birth or early after birth with dark, regular, and adherent scales of skin. Approximately 85%-90% of X-linked ichthyosis patients have large deletions of the STS gene and flanking sequences. Three patients have been identified with partial deletions of the gene. Two deletions have been found at the 3' extreme and the other one implicating exons 2-5. This study describes a novel partial deletion of the STS gene in an X-linked ichthyosis patient. The subject was classified through steroid sulfatase assay in leukocytes using 7-[3H] dehydroepiandrosterone sulfate as a substrate. Exons 1, 2, 5, and 7-10, and 3' flanking sequences DXS1131, DXS1133, DXS237, DXS1132, DXF22S1, and DXS278 of the STS gene were analyzed through polymerase chain reaction. The DNA analysis showed that exon 1 and 3' flanking sequences from DXS237 to DXS278 were present. In this study we report the fourth partial deletion of the STS gene and the first spanning exons 2-10 in X-linked ichthyosis patients. PMID- 10692124 TI - Interleukin-1 and cutaneous inflammation: a crucial link between innate and acquired immunity. AB - As our primary interface with the environment, the skin is constantly subjected to injury and invasion by pathogens. The fundamental force driving the evolution of the immune system has been the need to protect the host against overwhelming infection. The ability of T and B cells to recombine antigen receptor genes during development provides an efficient, flexible, and powerful immune system with nearly unlimited specificity for antigen. The capacity to expand subsets of antigen-specific lymphocytes that become activated by environmental antigens (memory response) is termed "acquired" immunity. Immunologic memory, although a fundamental aspect of mammalian biology, is a relatively recent evolutionary event that permits organisms to live for years to decades. "Innate" immunity, mediated by genes that remain in germ line conformation and encode for proteins that recognize conserved structural patterns on microorganisms, is a much more ancient system of host defense. Defensins and other antimicrobial peptides, complement and opsonins, and endocytic receptors are all considered components of the innate immune system. None of these, however, are signal-transducing receptors. Most recently, a large family of cell surface receptors that mediate signaling through the NF-kappaB transcription factor has been identified. This family of proteins shares striking homology with plant and Drosophila genes that mediate innate immunity. In mammals, this family includes the type I interleukin 1 receptor, the interleukin-18 receptor, and a growing family of Toll-like receptors, two of which were recently identified as signal-transducing receptors for bacterial endotoxin. In this review, we discuss how interleukin-1 links the innate and acquired immune systems to provide synergistic host defense activities in skin. PMID- 10692125 TI - A wavefront generator for complex pupil function synthesis and point spread function engineering. AB - We describe a simple method to produce an arbitrary complex optical field using a ferroelectric liquid crystal spatial light modulator. The system is configured so as to act as a pupil plane filter in a confocal microscope. The ability to tune the complex pupil function permits the system to be used both to modify the imaging performance by effectively engineering the point spread function as well as to remove optical aberrations present in the optical system. PMID- 10692126 TI - Image processing experiments for computer-based three-dimensional reconstruction of neurones from electron micrographs from serial ultrathin sections. AB - This study examined an image processing technique that uses a computer to reconstruct a three-dimensional image of neurones from electron micrographs of serial ultrathin sections. The major problems involved were: (a) a distortion of features in electron micrographs; (b) a significant change of cross-section features of neurones in electron micrographs of neighbouring sections; and (c) disagreement between the electron microscopic section face and the coordinate plane desired for the reconstruction. Electron micrographs of a retinal bipolar cell stained with a biotinylated tracer were used. We corrected the distortion of features by means of a warp, a widely used algorithm in morphing image processing. The change of features between neighbouring electron micrographs was minimized by filling the gaps with an interpolated image produced by a dissolve, another algorithm in morphing, as well as the warp. The distortion of the three dimensional reconstructed image made by piling up features was corrected by making the image with a wire frame model. Furthermore, in order to estimate a closed contour of features, an active contour model, Snakes, was applied to the electron microscope features. Snakes successfully detected the contour of the target feature, but in some electron microscope images broke into the target feature. PMID- 10692127 TI - ER-Tracker dye and BODIPY-brefeldin A differentiate the endoplasmic reticulum and golgi bodies from the tubular-vacuole system in living hyphae of Pisolithus tinctorius. AB - Two fluorochromes, ER-TrackerTM Blue-White DPX dye and the fluorescent brefeldin A (BFA) derivative, BODIPY-BFA, label the endoplasmic reticulum (ER) in hyphal tips of Pisolithus tinctorius and allow its differentiation from the tubular vacuole system at the light microscope level in living cells. The ER-Tracker dye labels a reticulate network similar in distribution to ER as seen in electron micrographs of freeze-substituted hyphae. BODIPY-BFA stains a thicker axially aligned structure with an expanded region at the apex, which is similar to that seen when hyphae are stained with ER-Tracker dye in the presence of unconjugated BFA. This structure is considered to be ER modified by BFA, a view supported by ultrastructural observations of the effect of BFA on the fungal ER. Both fluorescent probes also stain punctate structures, which are most likely to be Golgi bodies. Neither probe labels the tubular-vacuole system. PMID- 10692128 TI - Unsupervised approval criteria for automated EBSP investigation of deformed metals. AB - Unsupervised approval criteria have been investigated for orientations gathered from cold deformed samples (medium to high strain range) using the electron backscattering pattern technique. For such samples, the dislocation cell-size is on the order of the available step-size and pattern quality is generally low. Approval criteria for assessing the validity of measured orientations under these conditions were determined using, as a calibration, channel die cold deformed single crystals of stable orientations. In all cases, approval criteria based on an indexing confidence measure are found to be preferable. Different criteria are suggested, depending on whether the orientation data are subsequently to be used for texture analysis, or for a misorientation angle-based analysis. The latter is illustrated by an investigation of the number of deformation generated high angle boundaries introduced during a 90% cold reduction of a polycrystalline sample. PMID- 10692129 TI - Grain size distribution analysis in polycrystalline LiF thin films by mathematical morphology techniques on AFM images and X-ray diffraction data. AB - The influence of the deposition temperature on the grain size of polycrystalline lithium fluoride (LiF) thin films is studied using a mathematical morphology method. On atomic force microscopy images of the LiF surface, the grain sizes and shapes are determined by applying the watershed technique, together with a shape factor algorithm. Also, the domain size of the film structure, determined by an X ray diffraction data analysis, is compared and correlated with the mean grain size as a function of the deposition temperature. In both cases a linear increase with temperature and a very good agreement among the two structural parameters (grain and domain size) was found. PMID- 10692130 TI - Compact water-window transmission X-ray microscopy. AB - We demonstrate sub-100 nm resolution water-window soft X-ray full-field transmission microscopy with a compact system. The microscope operates at lambda = 3.37 nm and is based on a 100 Hz table-top regenerative debris-free droplet target laser-plasma X-ray source in combination with normal-incidence multilayer condenser optics for sample illumination. High-spatial-resolution imaging is performed with a 7.3% efficiency nickel zone plate and a 1024 x 1024 pixel CCD detector. Images of dry test samples are recorded with exposure times of a few minutes and show features smaller than 60 nm. PMID- 10692131 TI - Analysis of spherical aberration of a water immersion objective: application to specimens with refractive indices 1.33-1.40. AB - The method of using immersion medium to correct spherical aberration for water immersion objectives when the samples are not water is investigated. Spherical aberration is measured by an interferometer converted from a confocal microscope for samples with different refractive indices. When the proper refractive index of the immersion medium and thickness of cover slip are selected, the measured spherical aberration approaches zero. A theoretical model can be used for prediction of the immersion medium to correct spherical aberration for various samples. Using the thinnest available cover slip (100 microm), the zero spherical aberration condition can be applied to samples with refractive index as high as 1.40. Confocal images in the condition of almost no spherical aberration are included to demonstrate the improvement of axial resolution due to this correction. PMID- 10692132 TI - Retrospective shading correction based on entropy minimization. AB - Shading is a prominent phenomenon in microscopy, manifesting itself via spurious intensity variations not present in the original scene. The elimination of shading effects is frequently necessary for subsequent image processing tasks, especially if quantitative analysis is the final goal. While most of the shading effects may be minimized by setting up the image acquisition conditions carefully and capturing additional calibration images, object-dependent shading calls for retrospective correction. In this paper a novel method for retrospective shading correction is proposed. Firstly, the image formation process and the corresponding shading effects are described by a linear image formation model, which consists of an additive and a multiplicative parametric component. Secondly, shading correction is performed by the inverse of the image formation model, whose shading components are estimated retrospectively by minimizing the entropy of the acquired images. A number of tests, performed on artificial and real microscopical images, show that this approach is efficient for a variety of differently structured images and as such may have applications in and beyond the field of microscopical imaging. PMID- 10692133 TI - Iterative thresholding for segmentation of cells from noisy images. AB - We introduce an iterative thresholding algorithm for the segmentation of cells from noisy cell images. The thresholding image, which is initially a constant, changes iteratively with both the previous segmentation and image local activity. Experimental results for both synthesized and real cell images are provided to demonstrate the performance of the algorithm. PMID- 10692134 TI - An investigation of segmentation methods and texture analysis applied to tomographic images of human vertebral cancellous bone. AB - The goal of this study is to determine architectural and textural parameters on computed tomographic (CT) images, allowing us to explain the mechanical compressive properties of bone. Although the resolution (150 microm) is of the same order of magnitude as the trabecular thickness, this method enables the possibility of perfecting an in vivo peripheral CT system with an acceptable radiation dose for the patient. This study was performed on L2 vertebrae cancellous bone specimens taken after necropsy in 22 subjects aged 47-95 years (mean: 79 years). The segmentation process is a crucial point in the determination of accurate architectural parameters. In this paper the use of two different segmentation methods is investigated, based on an edge enhancement and a region growing approach. The images are compared and the architectural parameters extracted from the images segmented by both methods lead to a quantitative evaluation. The parameters are found to be globally robust towards the segmentation process, although some of them are much more sensitive to the approach used. Highly significant correlations (P < 0.0005) have been obtained between the two segmentation methods for all the parameters, with rho ranging from 0.70 to 0.93. In order to improve the assessment of bone architecture, texture analysis (run length method) was investigated. New features are obtained from an image reduced to 16 grey-levels. Textural parameters in addition to architectural parameters in a multivariate regression model increase significantly (P = 0.01) the prediction of the maximum compressive strength (variation of r2 from 0.75 up to 0.89). PMID- 10692136 TI - New Developments for the Journal of Neuroendocrinology. PMID- 10692135 TI - A simplified method of cleaning ixodid ticks for microscopy. AB - The cleaning of ixodid ticks for microscopy can be achieved quickly and efficiently using a combination of a wax solvent and an ultrasonic cleaner. The technique involves minimum handling of specimens, produces no detectable damage and is suitable for cleaning many specimens at the same time. PMID- 10692137 TI - Environmental oestrogens: a hazard to human reproductive health? PMID- 10692138 TI - Influence of neonatal rearing conditions on stress-induced adrenocorticotropin responses and norepinepherine release in the hypothalamic paraventricular nucleus. AB - Postnatal rearing conditions influence the development of hypothalamic-pituitary adrenal (HPA) responses to stress in the rat. Thus, postnatal handling dampens HPA responsivity to stress, while prolonged periods of maternal separation have the opposite effect. HPA responses to stress are initiated by the release of corticotropin-releasing factor and/or arginine vasopressin from the neurones of the paraventricular nucleus of the hypothalamus (PVNh). A major source of input to the PVNh arises from brainstem noradrenergic neurones with signalling occurring via alpha1 adrenoreceptors. We examined the noradrenergic response to stress in the PVNh in adult animals exposed to daily periods of handling or maternal separation over the first 2 weeks of life using microdialysis in conscious animals. Maternal separation increased, while handling greatly decreased and norepinepherine responses to restraint stress in the PVNh as compared to non-handled controls; the same pattern was observed for plasma adrenocorticotropic hormone (ACTH) responses to stress. Rearing condition did not affect either alpha1 or alpha2 receptor levels in the PVNh. However, alpha2 receptor binding levels in the noradrenergic cell body regions of the locus coeruleus and the n. tractus solitarius were significantly increased in handled animals. These alpha2 receptors are principally located on noradrenergic neurones (i.e. autoreceptors) and inhibit noradrenaline release at terminal sites. The effects on alpha2 receptor levels could serve as a mechanism for the differences in stress-induced noradrenaline levels in the PVNh and in HPA activity among handled vs non-handled and maternal separation animals. Thus, early life events may serve to influence the differentiation of noradrenergic neurones and thus alter HPA responses stress in adulthood. PMID- 10692139 TI - Effects of chronic stress on hypothalamic lnterleukin-1beta, interleukin-2, and gonadotrophin-releasing hormone gene expression in ovariectomized rats. AB - The influence of chronic stress on the expression of interleukin (IL)-1beta and IL-2 mRNAs in ovariectomized rat brains, and the physiological consequences of the expression of these cytokines on hypothalamic-pituitary-gonadal (HPG) activity were investigated. Using polymerase chain reaction (PCR)-assisted semiquantitative analysis, we demonstrated alterated expression of IL-1beta and IL-2 mRNA during repeated cold stress; the expression of both IL-beta and IL-2 mRNA increased in the medial preoptic area and ventromedial hypothalamus, and decreased in the lateral hypothalamic area. In the arcuate nucleus/median eminence, IL-2 mRNA expression was dramatically decreased, in contrast to the increase in IL-1beta mRNA expression. Concomitant analysis of GnRH mRNA expression indicated significant suppression of GnRH synthesis in the chronic phase, and a strong negative correlation with cytokine expression in the medial preoptic area. Similar results were obtained in intact females exposed to this stress. These results, together with previous pharmacological studies, suggest that chronic stress may induce reproductive dysfunction through the effects of stress-induced expression of endogenous cytokines. PMID- 10692140 TI - Interleukin-3 and interleukin-6 stimulate bovine adrenal cortisol secretion through different pathways. AB - Several interleukins have been reported to play a major role in the regulation of steroid secretion at all three levels of the hypothalamic-pituitary-adrenal axis. The objective of this study was to investigate the effect of interleukin-3 (IL-3) and interleukin-6 (IL-6) on cortisol secretion of bovine adrenocortical cells in primary culture under serum-free conditions. Both IL-3 and IL-6 stimulated basal cortisol secretion dose-dependently to a similar extent at a similar time course. After incubation with IL-3 or IL-6 at concentrations of 100 microg/l, a maximum 4.1-fold increase of the cortisol secretion was reached after 12 h (P<0.01). Coincubation of IL-3 and IL-6 (100 microg/l) revealed no significant synergism. To elucidate a possible involvement of arachidonic acid metabolites in the signal transduction, we coincubated IL-3 or IL-6 together with the cyclo-oxygenase inhibitor indometacin or the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA). Coincubation with indometacin completely abolished the stimulatory effect of IL-6 but had no effect on IL-3 stimulated cortisol secretion. In contrast, specific inhibition of the lipoxygenase system by nordihydroguaiaretic acid blocked IL-3 stimulated steroidogenesis while the effect of IL-6 was not affected. Neither IL-3 nor IL-6 altered cAMP levels significantly, whereas ACTH significantly induced cAMP levels in parallel to its steroidogenic effect. In conclusion, our data indicate that IL-3 and IL-6 stimulate the steroid secretion of bovine adrenocortical cells to a similar extent and with a similar time course. However, the effects of IL-3 and IL-6 are mediated through different, cAMP-independent pathways. While the stimulatory effect of IL-3 seems to be dependent on the lipoxygenase pathway, the effect of IL-6 on adrenocortical cortisol secretion is mediated through the cyclo-oxygenase pathway. PMID- 10692141 TI - Anatomically specific changes in the expression of somatostatin, growth hormone releasing hormone and growth hormone receptor mRNA in diabetic rats. AB - Growth hormone (GH) secretion is altered in poorly controlled diabetic animals. However, modifications in the hypothalamic neuropeptides that control GH secretion, somatostatin and GH-releasing hormone (GHRH), as well as changes in the sensitivity of the hypothalamus and pituitary to the feedback effects of GH, are less clear. We have used RNase protection assays and in-situ hybridization to address whether the mRNA expression of GH, somatostatin and GHRH, as well as of the GH receptor (GHR) in the hypothalamus and anterior pituitary, are altered in streptozotocin-induced diabetic rats. After induction of diabetes, rats were treated with insulin twice daily for 3 weeks to obtain either poorly controlled (mean plasma glucose >300 mg/dl) or well-controlled diabetic rats. Although no significant change in pituitary GH mRNA expression was found, the hypothalamic expression of GHRH and somatostatin mRNA was reduced in poorly-controlled diabetic rats and returned to control values with normalisation of plasma glucose concentrations (P<0.0001 and P<0.002, respectively). Somatostatin mRNA expression was reduced only in the central portion of the periventricular nucleus, with no change being seen in the other areas of the periventricular nucleus or in the arcuate, suprachiasmatic or paraventricular nuclei. A significant decline in GHRH mRNA expression was observed in both the arcuate nucleus and ventromedial hypothalamus. Anterior pituitary GHR mRNA expression was significantly reduced in both well and poorly-controlled diabetic rats, while there was no change in the hypothalamus. To examine whether the evolution time of the diabetes influences these parameters, in a subsequent experiment, diabetic rats received no insulin for 2 months. A significant decline in GHRH and somatostatin mRNA expression was also observed in these rats. In addition, pituitary GH mRNA expression declined significantly in long-term diabetic rats. These results demonstrate that: (1) the expression of both GHRH and somatostatin declines specifically in anatomical areas involved in anterior pituitary hormone control; (2) GHR mRNA expression is decreased in the pituitary of diabetic rats, but not in the hypothalamus, and does not return to control values with normalisation of mean blood glucose concentrations; and (3) the evolution time of the diabetes is important for detecting some changes, including the decrease in pituitary GH mRNA expression. PMID- 10692142 TI - Regulation of GABAA receptor by protein tyrosine kinases in frog pituitary melanotrophs. AB - The effects of protein tyrosine kinase (PTK) and PTK inhibitors on the GABAA receptor function were studied in cultured frog pituitary melanotrophs by using the patch-clamp technique. Extracellular application of the PTK inhibitors genistein (10-9 to 10-5 M) or lavendustin A (10-12 to 10-7 M) provoked a bell shaped potentiation of the whole-cell current induced by GABA (3x10-6 M). In contrast, at high concentrations, genistein (10-4 M) and lavendustin A (10-5 M) reversibly reduced the GABA-evoked current. Daidzein and lavendustin B, the inactive analogs of genistein and lavendustin A, respectively, did not modify the current induced by GABA. In the inside-out configuration, bath application of the recombinant PTK pp60c-src (75 U/ml) inhibited the GABA-activated chloride current, and the inhibitory effect of pp60c-src was prevented by genistein (10-7 M). Immunoblotting revealed that genistein, at doses of 10-7 M or 10-4 M, markedly inhibited tyrosine phosphorylation of the beta2/beta3 subunits of the GABAA receptor. Extracellular application of the PKA activator Bt2cAMP (10-3 M), the PKA/PKC inhibitor H7 (10-5 M) and the Cam KII inhibitor W7 (10-5 M) reversibly diminished the whole-cell GABA-induced current. Internal application of H7 and W7 (10-4 M) did not modify the dose-dependent effects of genistein. Internal application of sodium orthovanadate (10-4 M), a protein tyrosine phosphatase inhibitor, decreased the GABA-evoked current and markedly reduced the potentiating effect of genistein. The present study provides the first evidence that, in frog pituitary melanotrophs, the GABAA receptor is phosphorylated at least on its beta2/beta3 subunits by an endogenous PTK. Our data also demonstrate that tyrosine phosphorylation exerts an inhibitory effect on GABAA receptor function. PMID- 10692143 TI - The influence of gonadal steroids on pre-pro melanin-concentrating hormone mRNA in female rats. AB - Melanin-concentrating hormone (MCH) may have a regulatory role in the control of luteinizing hormone (LH) release. We have investigated if gonadal steroids induce changes in the expression of pre-pro MCH (ppMCH) that are associated with changes in the pattern of LH release. Using quantitative in-situ hybridization histochemistry we have determined the effect of administration of either oestradiol benzoate (5 microg/rat) or oestradiol benzoate followed 44 or 48 h later by progesterone (0.5 mg/rat) to ovariectomized rats on the expression of ppMCH in the medial and lateral zona incerta and the lateral hypothalamus. The prevalence of ppMCH transcripts in the intact female rat at 12.00 and 19.00 h on proestrus and the first day of dioestrus was also examined. Oestrogen reduced the intensity of hybridization signal for ppMCH mRNA and this was associated with both a decrease in the number of cells in which the message was detected in the medial zona incerta and a negative feedback effect on LH release in ovariectomized rats. Progesterone administration to oestradiol benzoate-primed rats did not alter the reduced expression in the medial zona incerta in spite of its positive feedback effect on LH release. We suggest that progesterone may act only on post-translational events. Expression in the MCH cell bodies of the lateral zona incerta were not affected but there was a transient decrease 4 h after progesterone treatment in the oestradiol benzoate-primed rats in expression in the lateral hypothalamus. No changes in ppMCH mRNA were seen in intact animals on proestrus or the first day of dioestrus indicating that gonadal steroids are not important in the modulation of ppMCH gene expression during the oestrous cycle. In other steroid-dependent physiological situations, however, oestrogen may influence the expression of ppMCH in a subpopulation of cell bodies in the medial zona incerta. PMID- 10692144 TI - Potentiation effect of vasopressin on melatonin secretion as determined by trans pineal microdialysis in the Rat. AB - The mammalian pineal gland is known to receive a noradrenergic innervation originating from the superior cervical ganglion which corresponds to the primary regulatory input for melatonin synthesis. However, many peptidergic fibers containing peptides such as vasopressin and oxytocin have also been found in the rat pineal gland. The present study was performed to investigate the possible role of vasopressin and oxytocin on melatonin secretion in vivo. Therefore, both neuropeptides were delivered for 2 h through a trans-pineal microdialysis probe directly into the gland at different times during the nocturnal phase of the light:dark cycle. At the same time pineal dialysates were collected continuously. Melatonin concentrations were measured by radioimmunoassay. Melatonin synthesis potentiation was achieved when vasopressin was infused locally in the pineal, during the onset of nocturnal melatonin secretion. In order to assess the possible role of a physiological increase of endogenous circulating vasopressin on pineal metabolism, melatonin synthesis was recorded in the same animals before and after a prolonged dehydration period. Night time melatonin concentration was increased after the water deprivation vs control conditions. Contrary to that, oxytocin seems not to affect pineal metabolism in the rat since no significant change was observed on melatonin secretion in response to a local oxytocin infusion. These results show that vasopressin can modulate melatonin synthesis in the rat pineal whereas no effect was obtained with oxytocin, at least under the present experimental conditions. PMID- 10692145 TI - Metabolic influences on circadian rhythmicity in Siberian and Syrian hamsters exposed to long photoperiods. AB - Calorie restriction and other situations of reduced glucose availability in rodents alter the entraining effects of light on the circadian pacemaker located in the suprachiasmatic nuclei. Siberian and Syrian hamsters are photoperiodic species that are sexually active when exposed to long summer-like photoperiods, while both species show opposite changes in body mass when transferred from long to short or short to long days. Because metabolic cues may fine tune the photoperiodic responses via the suprachiasmatic nuclei, we tested whether timed calorie restriction can alter the photic synchronization of the light-entrainable pacemaker in these two hamster species exposed to long photoperiods. Siberian and Syrian hamsters were exposed to 16 h:8 h light:dark cycles and received daily hypocaloric (75% of daily food intake) or normocaloric diet (100% of daily food intake) 4 h after light onset. Four weeks later, hamsters were transferred to constant darkness and fed ad libitum. The onset of the nocturnal pattern of locomotor activity was phase advanced by 1.5 h in calorie-restricted Siberian hamsters, but not in Syrian hamsters. The lack of phase change in calorie restricted Syrian hamsters was also observed in individuals exposed to 14 h:10 h dim light:dark cycles and fed with lower hypocaloric food (i.e. 60% of daily food intake) 2 h after light onset. Moreover, in hamsters housed in constant darkness and fed ad lib., light-induced phase shifts of the locomotor activity in Siberian hamsters, but not in Syrian hamsters were significantly reduced when glucose utilization was blocked by pretreatment with 500 mg/kg i.p. 2-deoxy-D-glucose. Taken together, these results show that the photic synchronization of the light entrainable pacemaker can be modulated by metabolic cues in Siberian hamsters, but not in Syrian hamsters maintained on long days. PMID- 10692146 TI - Differential effects of placental restriction on IGF-II, ACTH receptor and steroidogenic enzyme mRNA levels in the foetal sheep adrenal. AB - We have investigated the effects of restriction of placental growth on foetal adrenal growth and adrenal expression of mRNAs for Insulin-like Growth Factor II (IGF-II), the IGF binding protein IGFBP-2, Steroidogenic Factor 1 (SF-1) and adrenocorticotrophic hormone (ACTH) receptor (ACTH-R) and the steroidogenic cytochrome P-450 enzymes: cholesterol side chain cleavage (CYP11A1), 17alpha hydroxylase (CYP17) and 21-hydroxylase (CYP21A1); and 3beta-hydroxysteroid dehydrogenase/Delta5Delta4 isomerase (3betaHSD). Endometrial caruncles were removed from non-pregnant ewes before mating (placental restriction group; PR). The total adrenal: foetal weight ratio was higher in PR (n=6 foetuses) than in control foetuses (n=6 foetuses). There was no difference in plasma ACTH concentrations between the PR and control foetuses between 130 and 140 days gestation. Adrenal IGF-II mRNA levels were lower (P<0.05) in the PR group, however, adrenal IGFBP-2 mRNA levels were not different between the PR and control groups. Adrenal ACTH-R mRNA levels were also lower whilst CYP11A1 mRNA levels were increased (P<0.005) in the PR group. We conclude that foetal adrenal growth and steroidogenesis are stimulated as a consequence of foetal growth restriction and that factors other than ACTH are important in foetal adrenal activation during chronic, sustained hypoxaemia. PMID- 10692147 TI - Thyroid hormone acts centrally to programme photostimulated male american tree sparrows (Spizella arborea) for vernal and autumnal components of seasonality. AB - Thyroid hormone and long days interact to programme American tree sparrows (Spizella arborea) for seasonality (i.e. thyroid hormone-dependent photoperiodic gonadal growth, photorefractoriness, and postnuptial moult). This study explored in radiothyroidectomized (THX) males given thyroid hormone replacement therapy whether thyroid hormone acts within the brain and, additionally, the identity of the putative tissue-active thyroid hormone. The minimum dose (30 ng) of L thyroxine (T4) that restored all components of seasonality when given i.c.v. daily during the first 21 days of photostimulation restored no component of seasonality when given s.c. The same dose of L-triiodothyronine (T3) also was ineffective when administered s.c., but restored photoperiodic testicular growth (though neither photorefractoriness nor postnuptial moult) when admiministered i.c.v. Three of seven birds given a 10-fold lower dose of T4 (3 ng) exhibited thyroid hormone-dependent photoperiodic testicular growth, albeit damped. The other four birds given 3 ng T4 and all birds given 3 ng T3 responded like THX controls, exhibiting only slight thyroid hormone-independent photoperiodic testicular growth. The highest dose (300 ng) of T3 restored all components of seasonality only when administered i.c.v. daily during the first 49 days of photostimulation. This demonstration in American tree sparrows is the first in any species that the thyroid-dependent transition from the breeding season to the non-breeding season can be effected by T3. The same dose of reverse T3 administered daily over the same 49 days restored photoperiodic testicular growth in only half of 10 subjects and photorefractoriness and moult in none. Collectively, the data support the hypothesis that thyroid hormone acts centrally to programme photostimulated male American tree sparrows for all components of seasonality. The most parsimonious interpretation of the data, including the threshold-like effect of 3 ng T4, favours T4 as the tissue-active thyroid hormone for vernal as well as autumnal events, but does not entirely exclude T3. PMID- 10692148 TI - Families of transmembrane sugar transport proteins. AB - We describe here 20 families of secondary (pmf-driven) carriers which, in addition to nine families within the ATP-dependent ABC superfamily, and seven families of Gram-negative bacterial outer membrane porins, largely account for the stereospecific transport of sugars and their derivatives into and out of all living cells on earth. Family characteristics as well as struc-tural and functional properties of the family constituents are described. By reference to our website (http://www-biology.ucsd.edu/ approximately msaier/transport/), phylogenetic relationships, detailed substrate specificity information and both primary and secondary references are also available. This review provides a comprehensive guide to the diversity of carriers that mediate the transport of sugar-containing molecules across cell and organellar membranes. PMID- 10692149 TI - Outer-membrane phospholipase A: known structure, unknown biological function. AB - Outer-membrane phospholipase A (OMPLA) is one of the few enzymes present in the outer membrane of Gram-negative bacteria. The enzymatic activity of OMPLA is strictly regulated to prevent uncontrolled breakdown of the surrounding phospholipids. The activity of OMPLA can be induced by membrane perturbation and concurs with dimerization of the enzyme. The recently elucidated crystal structures of the inactive, monomeric and an inhibited dimeric form of the enzyme provide detailed structural insight into the functional properties of the enzyme. OMPLA is a serine hydrolase with a unique Asn-156-His-142-Ser-144 catalytic triad. Only in the dimeric state, complete substrate binding pockets and functional oxyanion holes are formed. A model is proposed for the activation of OMPLA in which membrane perturbation causes the formation of non-bilayer structures, resulting in the presentation of phospholipids to the active site of OMPLA and leading to the formation of the active dimeric species. Possible roles for OMPLA in maintaining the cell envelope integrity and in pathogenicity are discussed. PMID- 10692150 TI - Lipopolysaccharide core phosphates are required for viability and intrinsic drug resistance in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is an opportunistic pathogen that is notorious for its intrinsic drug resistance. We have used chemical and genetic techniques to characterize three putative kinase genes that are involved in the addition of phosphate to the inner core region of P. aeruginosa lipopolysaccharide. The first gene is a waaP homologue, whereas the other two (wapP and wapQ) are unique to P. aeruginosa. Repeated attempts using a variety of membrane-stabilizing conditions to generate waaP:Gm (Gm, gentamicin) or wapP:Gm mutants were unsuccessful. We were able to generate a chromosomal waaP mutant that had a wild-type copy of either waaPPa or waaPEc in trans, but were unable to cure this plasmid-borne copy of the gene. These results are consistent with the fact that P. aeruginosa mutants lacking inner core heptose (Hep) or phosphate have never been isolated and demonstrate the requirement of Hep-linked phosphate for P. aeruginosa viability. A wapQ:Gm mutant was isolated and it had an unaltered minimum inhibitory concentration (MIC) for novobiocin and only a small decrease in the MIC for sodium dodecyl sulphate (SDS), suggesting that the loss of a phosphate group transferred by WapQ may only be having a small impact on outer-membrane permeability. Nuclear magnetic resonance and methylation linkage analysis showed that WaaPPa could add one phosphate to O4 of HepI in a Salmonella typhimurium waaP mutant. The expression of WaaPPa increased the outer-membrane integrity of these complemented mutants, as evidenced by 35-fold and 75-fold increases in the MIC for novobiocin and SDS respectively. The S. typhimurium waaP mutant transformed with both waaP and wapP had over 250-fold and 1000-fold increases, respectively, in these MICs. The inner core phosphates of P. aeruginosa appear to be playing a key role in the intrinsic drug resistance of this bacterium. PMID- 10692151 TI - DNA methylation-dependent regulation of pef expression in Salmonella typhimurium. AB - Plasmid-encoded fimbriae (Pef) expressed by Salmonella typhimurium mediate adhesion to mouse intestinal epithelium. The pef operon shares features with the Escherichia coli pyelonephritis-associated pilus (pap) operon, which is under methylation-dependent transcriptional regulation. These features include conserved DNA GATC box sites in the upstream regulatory region as well as homologues of the PapI and PapB regulatory proteins. Unlike Pap fimbriae, which are expressed in a variety of laboratory media, Pef fimbriae were expressed only in acidic, rich broth under standing culture conditions. Analysis of S. typhimurium grown under these conditions indicated that Pef production was regulated by a phase variation mechanism, in which the bacterial population was skewed between fimbrial expression (phase ON) and non-expression (phase OFF) states. Leucine-responsive regulatory protein (Lrp) and DNA adenine methylase (Dam) were required for pef transcription. In contrast, the histone-like protein (H-NS) and the stationary-phase sigma factor (RpoS) repressed pef transcription. Methylation of the pef GATC II site appeared to be required for pef fimbrial expression based on analysis of a GCTC II mutant that did not express Pef fimbriae. Analysis of the DNA methylation states of pef GATC sites indicated that, under acidic growth conditions, which induced Pef production, most GATC I sites were non-methylated, whereas GATC II and GATC X were predominantly methylated. The methylation protection at GATC I and GATC II was dependent upon Lrp and was modulated by PefI. Together, these results indicate that Pef production is regulated by DNA methylation, which is the first example of methylation-dependent gene regulation outside of E. coli. PMID- 10692152 TI - Phosphorylation of the flagellar regulatory protein FlrC is necessary for Vibrio cholerae motility and enhanced colonization. AB - The human pathogen Vibrio cholerae specifically expresses virulence factors within the host, including cholera toxin (CT) and the toxin co-regulated pilus (TCP), which allow it to colonize the intestine and cause disease. V. cholerae is a highly motile organism by virtue of a polar flagellum, and motility has been inferred to be an important aspect of virulence, yet the exact role of motility in pathogenesis has remained undefined. The two-component regulatory system FlrB/FlrC is required for polar flagellar synthesis; FlrC is a sigma54-dependent transcriptional activator. We demonstrate that the transcriptional activity of FlrC affects both motility and colonization of V. cholerae. In a purified in vitro reaction, FlrB transfers phosphate to the wild-type FlrC protein, but not to a mutant form in which the aspartate residue at amino acid position 54 has been changed to alanine (D54A), consistent with this being the site of phosphorylation of FlrC. The wild-type FlrC protein, but not the D54A protein, activates sigma54-dependent transcription in a heterologous system, demonstrating that phospho-FlrC is the transcriptionally active form. A V. cholerae strain containing a chromosomal flrCD54A allele did not synthesize a flagellum and had no detectable levels of transcription of the critical sigma54-dependent flagellin gene flaA. The V. cholerae flrCD54A mutant strain was also defective in its ability to colonize the infant mouse small intestine, approximately 50-fold worse than an isogenic wild-type strain. Another mutation of FlrC (methionine 114 to isoleucine; M114I) confers constitutive transcriptional activity in the absence of phosphorylation, but a V. cholerae flrCM114I mutant strain, although flagellated and motile, was also defective in its ability to colonize. The strains carrying D54A or M114I mutant FlrC proteins expressed normal levels of CT and TCP under in vitro inducing conditions. Our results show that FlrC 'locked' into either an inactive (D54A) or an active (M114I) state results in colonization defects, thereby demonstrating a requirement for modulation of FlrC activity during V. cholerae pathogenesis. Thus, the sigma54-dependent transcriptional activity of the flagellar regulatory protein FlrC contributes not only to motility, but also to colonization of V. cholerae. PMID- 10692153 TI - Two roles for integration host factor at an enhancer-dependent nifA promoter. AB - Control of transcription in prokaryotes often involves direct contact of regulatory proteins with RNA polymerase. For the sigma54 RNA polymerase, regulatory proteins bound to distally located enhancers engage the polymerase via DNA looping. The sigma54-dependent nifA promoter of Herbaspirillum seropedicae (Hs) is activated under nitrogen-limiting growth conditions. Potential enhancers for the nitrogen control activators NTRC and NIFA and binding sites for integration host factor (IHF) and sigma54-holoenzyme were identified. DNA footprinting experiments showed that these sites functioned for protein binding. Their involvement in the promoter regulation was explored. In vitro, activation of the Hs nifA promoter by NTRC is stimulated by the DNA bending protein IHF. In marked contrast, activation by NIFA is greatly reduced by IHF, thus diminishing potentially destabilizing autoactivation of the nifA promoter by NIFA. Additionally, high levels of NIFA appear to limit NTRC-dependent activation. This inhibition is IHF dependent. Therefore, IHF acts positively and negatively at the nifA promoter to restrict transcription activation to NTRC and one signal transduction pathway. PMID- 10692154 TI - A new mechanism for the control of a prokaryotic transcriptional regulator: antagonistic binding of positive and negative effectors. AB - MalT, the transcriptional activator of the Escherichia coli maltose regulon, self associates, binds promoter DNA and activates initiation of transcription only in the presence of ATP and maltotriose, the inducer. In vivo studies have revealed that MalT action is negatively controlled by the MalY protein. Using a biochemical approach, we analyse here the mechanism whereby MalY represses MalT activity. We show that MalY inhibits transcription activation by MalT in a purified transcription system. In vitro, a constitutive MalT variant (which is partially active in the absence of maltotriose) is less sensitive than wild-type MalT to repression by MalY, as observed in vivo. We demonstrate that MalY forms a complex with MalT only in the absence of maltotriose and that, conversely, MalY inhibits maltotriose binding by MalT. Together, these results establish that MalY acts directly upon MalT without the help of any factor, and that MalY is a negative effector of MalT competing with the inducer for MalT binding. PMID- 10692155 TI - In vivo and in vitro studies of transmembrane beta-strand deletion, insertion or substitution mutants of the Escherichia coli K-12 maltoporin. AB - LamB of Escherichia coli K12, also called maltoporin, is an outer membrane protein, which specifically facilitates the diffusion of maltose and maltodextrin through the bacterial outer membrane. Each monomer is composed of an 18-stranded antiparallel beta-barrel. In the present work, on the basis of the known X-ray structure of LamB, the effects of modifications of the beta-barrel domain of maltoporin were studied in vivo and in vitro. We show that: (i) the substitution of the pair of strands beta13-beta14 of the E. coli maltoporin with the corresponding pair of strands from the functionally related maltoporin of Salmonella typhimurium yielded a protein active in vivo and in vitro; and (ii) the removal of one pair of beta-strands (deletion beta13-beta14) from the E. coli maltoporin, or its replacement by a pair of strands from the general porin OmpF of E. coli, leads to recombinant proteins that lost in vivo maltoporin activities but still kept channel formation and carbohydrate binding in vitro. We also inserted into deletion beta13-beta14 the portion of the E. coli LamB protein comprising strands beta13 to beta16. This resulted in a protein expected to have 20 beta-strands and which completely lost all LamB-specific activities in vivo and in vitro. PMID- 10692156 TI - A positive feedback mechanism controls expression of AlkS, the transcriptional regulator of the Pseudomonas oleovorans alkane degradation pathway. AB - The AlkS regulator, encoded by the alkS gene of the Pseudomonas oleovorans OCT plasmid, activates the expression of a set of enzymes that allow assimilation of alkanes. We show that the AlkS protein regulates, both negatively and positively, the expression of its own gene. In the absence of alkanes, alkS is expressed from promoter PalkS1, which is recognized by sigmaS-RNA polymerase, and whose activity is very low in the exponential phase of growth and considerably higher in stationary phase. AlkS was found to downregulate this promoter, limiting expression of alkS in stationary phase when alkanes were absent. In the presence of alkanes, AlkS repressed PalkS1 more strongly and simultaneously activated a second promoter for alkS, named PalkS2, located 38 bp downstream from PalkS1. Activation of PalkS2 allowed efficient transcription of alkS when alkanes were present. Transcription from PalkS2 was modulated by catabolite repression when cells were provided with a preferred carbon source. We propose that the expression of alkS is regulated by a positive feedback mechanism, which leads to a rapid increase in alkS transcription when alkanes are present. This mechanism should allow a rapid induction of the pathway, as well as a fast switch-off when alkanes are depleted. An improved model for the regulation of the pathway is proposed. PMID- 10692157 TI - CtsR controls class III heat shock gene expression in the human pathogen Listeria monocytogenes. AB - Stress proteins play an important role in virulence, yet little is known about the regulation of stress response in pathogens. In the facultative intracellular pathogen Listeria monocytogenes, the Clp ATPases, including ClpC, ClpP and ClpE, are required for stress survival and intracellular growth. The first gene of the clpC operon of L. monocytogenes encodes a homologue of the Bacillus subtilis CtsR repressor of stress response genes. An L. monocytogenes ctsR-deleted mutant displayed enhanced survival under stress conditions (growth in the presence of 2% NaCl or at 42 degrees C), but its level of virulence in the mouse was not affected. The virulence of a wild-type strain constitutively expressing CtsR is significantly attenuated, presumably because of repression of the stress response. Regulation of the L. monocytogenes clpC, clpP and clpE genes was investigated using transcriptional fusions in B. subtilis as a host. The L. monocytogenes ctsR gene was placed under the control of an inducible promoter, and regulation by CtsR and heat shock was demonstrated in vivo in B. subtilis. The purified CtsR protein of L. monocytogenes binds specifically to the clpC, clpP and clpE regulatory regions, and the extent of the CtsR binding sites was defined by DNase I footprinting. Our results demonstrate that this human pathogen possesses a CtsR regulon controlling class III heat shock genes, strikingly similar to that of the saprophyte B. subtilis. This is the first description of a stress response regulatory gene in a pathogen. PMID- 10692158 TI - The asgE locus is required for cell-cell signalling during Myxococcus xanthus development. AB - In response to starvation, Myxococcus xanthus undergoes a multicellular developmental process that produces a dome-shaped fruiting body structure filled with differentiated cells called myxospores. Two insertion mutants that block the final stages of fruiting body morphogenesis and reduce sporulation efficiency were isolated and characterized. DNA sequence analysis revealed that the chromosomal insertions are located in open reading frames ORF2 and asgE, which are separated by 68 bp. The sporulation defect of cells carrying the asgE insertion can be rescued phenotypically when co-developed with wild-type cells, whereas the sporulation efficiency of cells carrying the ORF2 insertion was not improved when mixed with wild-type cells. Thus, the asgE insertion mutant appears to belong to a class of developmental mutants that are unable to produce cell cell signals required for M. xanthus development, but they retain the ability to respond to them when they are provided by wild-type cells. Several lines of evidence indicate that asgE cells fail to produce normal levels of A-factor, a cell density signal. A-factor consists of a mixture of heat-stable amino acids and peptides, and at least two heat-labile extracellular proteases. The asgE mutant yielded about 10-fold less heat-labile A-factor and about twofold less heat-stable A-factor than wild-type cells, suggesting that the primary defect of asgE cells is in the production or release of heat-labile A-factor. PMID- 10692159 TI - Interactions between Pho85 cyclin-dependent kinase complexes and the Swi5 transcription factor in budding yeast. AB - Pho85 is a cyclin-dependent protein kinase (Cdk) in budding yeast with roles in cell metabolism and cell cycle progression. Activation of Pho85 occurs through association with Pho85 cyclins (Pcls), of which 10 are known. When complexed with the G1 cyclins, Pcl1 and Pcl2, Pho85 is required for cell cycle progression in the absence of the Cdc28-dependent cyclins, Cln1 and Cln2. To identify potential targets of Pcl2-Pho85, we performed a two-hybrid screen using the Pcl2 cyclin as bait and recovered the transcription factor Swi5 as a Pcl2-interacting protein. We performed both biochemical and genetic tests to discover the biological significance of the interaction between Pcl2 and Swi5 seen in the two-hybrid assay. We found that Swi5 interacts in vitro with Pho85 cyclins and is phosphorylated in vitro by the Pho80-Pho85 kinase. We discovered that a subset of genes that are controlled by Swi5 and a homologous transcription factor, Ace2, was misregulated in a pho85 deletion strain; expression of the ASH1 and CTS1 genes was reduced in an ace2 deletion strain, whereas expression of both genes was increased in an ace2Delta pho85Delta double mutant. We also found that overexpression of SWI5 caused cell lethality in a pho85 deletion strain. Our results are consistent with misregulation of Swi5 activity in vivo in the absence of Pho85 and implicate Swi5 as a potential substrate of Pho85 cyclin-dependent kinase complexes. PMID- 10692160 TI - IHF redistributes bound initiator protein, DnaA, on supercoiled oriC of Escherichia coli. AB - In Escherichia coli, initiation of chromosome replication requires that DnaA binds to R boxes (9-mer repeats) in oriC, the unique chromosomal replication origin. At the time of initiation, integration host factor (IHF) also binds to a specific site in oriC. IHF stimulates open complex formation by DnaA on supercoiled oriC in cell-free replication systems, but it is unclear whether this stimulation involves specific changes in the oriC nucleoprotein complex. Using dimethylsulphate (DMS) footprinting on supercoiled oriC plasmids, we observed that IHF redistributed prebound DnaA, stimulating binding to sites R2, R3 and R5(M), as well as to three previously unidentified non-R sites with consensus sequence (A/T)G(G/C) (A/T)N(G/C)G(A/T)(A/T)(T/C)A. Redistribution was dependent on IHF binding to its cognate site and also required a functional R4 box. By reducing the DnaA level required to separate DNA strands and trigger initiation of DNA replication at each origin, IHF eliminates competition between strong and weak sites for free DnaA and enhances the precision of initiation synchrony during the cell cycle. PMID- 10692161 TI - Involvement of differential efficiency of transcription by esigmas and esigma70 RNA polymerase holoenzymes in growth phase regulation of the Escherichia coli osmE promoter. AB - Transcription of the gene osmE of Escherichia coli is inducible by elevated osmotic pressure and during the decelerating phase of growth. osmE expression is directed by a single promoter, osmEp. Decelerating phase induction of osmEp is dependent on the sigmas (RpoS) factor, whereas its osmotic induction is independent of sigmas. Purified Esigmas and Esigma70 were both able to transcribe osmEp in vitro on supercoiled templates. In the presence of rpoD800, a mutation resulting in a thermosensitive sigma70 factor, a shift to non-permissive temperature abolished induction of osmEp after an osmotic shock during exponential phase, but did not affect the decelerating phase induction. Point mutations affecting osmEp activity were isolated. Down-promoter mutations decreased transcription in both the presence and the absence of sigmas, indicating that the two forms of RNA polymerase holoenzyme recognize very similar sequence determinants on the osmE promoter. Three up-promoter mutations brought osmEp closer to the consensus of Esigma70-dependent promoters. The two variant promoters exhibiting the highest efficiency became essentially independent of sigmas in vivo. Our data suggest that Esigmas transcribes wild-type osmEp with a higher efficiency than Esigma70. A model in which an intrinsic differential recognition contributes to growth phase-dependent regulation is proposed. Generalization of this model to other sigmas-dependent promoters is discussed. PMID- 10692162 TI - Identification of novel VirR/VirS-regulated genes in Clostridium perfringens. AB - Novel genes that are regulated in Clostridium perfringens by the two-component regulatory system, VirR/VirS, were identified using a differential display method. A plasmid library was constructed from C. perfringens chromosomal DNA, and the plasmids were hybridized with cDNA probes prepared from total RNA of wild type strain 13 and its virR mutant derivative TS133. Three clones were identified that carry newly identified VirR/VirS-regulated genes, two of which were positively regulated and one of which was negatively regulated. Genes located on the identified clones were deduced by nucleotide sequencing, and the target genes of the VirR/VirS system were identified with a set of Northern hybridizations. A 4.9 kb mRNA transcribing the metB (cystathionine gamma-synthase), cysK (cysteine synthase) and ygaG (hypothetical protein) genes was negatively regulated, whereas 1.6 and 6.0 kb transcripts encoding ptp (protein tyrosine phosphatase) and cpd (2',3'-cyclic nucleotide 2'-phosphodiesterase) respectively, were shown to be positively regulated by the VirR/VirS system. The other gene, hyp7, whose transcript was positively regulated by the VirR/VirS system, was shown to activate the transcription of the colA (kappa-toxin) and plc (alpha-toxin) genes, but not the pfoA (theta-toxin) gene in C. perfringens. These results suggested that the global regulatory system VirR/VirS could regulate various genes, other than toxin genes, both positively and negatively and that the hyp7 gene might encode a novel regulatory factor for toxin production in C. perfringens. PMID- 10692163 TI - Evolutionary conservation of prion-forming abilities of the yeast Sup35 protein. AB - Saccharomyces cerevisiae prion [PSI ] is a self-propagating isoform of the eukaryotic release factor eRF3 (Sup35p). Sup35p consists of the evolutionary conserved release factor domain (Sup35C) and two evolutionary variable regions - Sup35N, which serves as a prion-forming domain in S. cerevisiae, and Sup35M. Here, we demonstrate that the prion form of Sup35p is not observed among industrial and natural strains of yeast. Moreover, the prion ([PSI + ]) state of the endogenous S. cerevisiae Sup35p cannot be transmitted to the next generations via heterologous Sup35p or Sup35NM, originating from the distantly related yeast species Pichia methanolica. This suggests the existence of a 'species barrier' in yeast prion conversion. However, the chimeric Sup35p, containing the Sup35NM region of Pichia, can be turned into a prion in S. cerevisiae by overproduction of the identical Pichia Sup35NM. Therefore, the prion-forming potential of Sup35NM is conserved in evolution. In the heterologous system, overproduction of Pichia Sup35p or Sup35NM induced formation of the prion form of S. cerevisiae Sup35p, albeit less efficiently than overproduction of the endogenous Sup35p. This implies that prion induction by protein overproduction does not require strict correspondence of the 'inducer' and 'inducee' sequences, and can overcome the 'species barrier'. PMID- 10692164 TI - Phase variation of Ag43 in Escherichia coli: Dam-dependent methylation abrogates OxyR binding and OxyR-mediated repression of transcription. AB - It has been shown previously that phase variation of the outer membrane protein Antigen43 (Ag43) of Escherichia coli requires the DNA-methylating enzyme deoxyadenosine methyltransferase (Dam) and the global regulator OxyR. In this study, we analysed the regulation of the Ag43 encoding gene (agn) using isolates containing a fusion of the agn regulatory region to the reporter gene lacZ. Our results indicate that phase variation of Ag43 is regulated at the level of transcription. Repression of agn'-lacZ transcription required OxyR, whereas activation required Dam. The regulatory region of agn contains three GATC sequences, which are target sites for Dam-dependent methylation. In vivo, the methylation state of these GATC sequences correlated with the transcription state of agn'-lacZ. These GATC sequences were not protected from Dam-dependent methylation in an oxyR background, suggesting that OxyR binding results in Dam dependent methylation protection in OFF cells. In vitro, both oxidized OxyR and OxyR(C199S), which is locked in the reduced conformation, bound to the agn regulatory region, but methylation of the three GATC sequences abrogated this binding. In vivo, OxyR(C199S) was sufficient to repress Ag43 transcription. Our data support a model in which OxyR-mediated repression of transcription is alleviated by methylation of three GATC sequences in its binding site. In addition, we show that, in an oxyR background, Dam was still required for full activation, suggesting that the model concerning the role of Dam in agn regulation is incomplete. These results show that Dam-dependent phase variation in E. coli is not limited to the previously identified regulatory system of the family of pap-like fimbrial operons. PMID- 10692165 TI - Identification of a unique domain essential for Escherichia coli DNA topoisomerase III-catalysed decatenation of replication intermediates. AB - A 17-amino-acid residue domain has been identified in Escherichia coli DNA topoisomerase III (Topo III) that is essential for Topo III-mediated resolution of DNA replication intermediates in vitro. Deletion of this domain reduced Topo III-catalysed resolution of DNA replication intermediates and decatenation of multiply linked plasmid DNA dimers by four orders of magnitude, whereas reducing Topo III-catalysed relaxation of negatively supercoiled DNA substrates only 20 fold. The presence of this domain has been detected in multiple plasmid-encoded topoisomerases, raising the possibility that these enzymes may also be decatenases. PMID- 10692166 TI - Delineation of pilin domains required for bacterial association into microcolonies and intestinal colonization by Vibrio cholerae. AB - The toxin-co-regulated pilus (TCP), a type 4 pilus that is expressed by epidemic strains of Vibrio cholerae O1 and O139, is required for colonization of the human intestine. The TCP structure is assembled as a polymer of repeating subunits of TcpA pilin that form long fibres, which laterally associate into bundles. Previous passive immunization studies have suggested that the C-terminal region of TcpA is exposed on the surface of the pilus fibre and has a critical role in mediating the colonization functions of TCP. In the present study, we have used site-directed mutagenesis to delineate two domains within the C-terminal region that contribute to TCP structure and function. Alterations in the first domain, termed the structural domain, result in altered pilus stability or morphology. Alterations in the second domain, termed the interaction domain, affect colonization and/or infection by CTX-bacteriophage without affecting pilus morphology. In vitro and in vivo analyses of the tcpA mutants revealed that a major function of TCP is to mediate bacterial interaction through direct pilus pilus contact required for microcolony formation and productive intestinal colonization. The importance of this function is supported by the finding that intragenic suppressor mutations that restore colonization ability to colonization deficient mutants simultaneously restore pilus-mediated bacterial interactions. The alterations resulting from the suppressor mutations also provide insight into the molecular interactions between pilin subunits within and between pilus fibres. PMID- 10692167 TI - A novel mechanism for control of antigenic variation in the haemagglutinin gene family of mycoplasma synoviae. AB - High-frequency phase and antigenic variation of homologous lipoprotein haemagglutinins has been seen in both the major avian mycoplasma pathogens, Mycoplasma synoviae and Mycoplasma gallisepticum. The expression and, hence, antigenic variation of the pMGA gene family (encoding these lipoproteins in M. gallisepticum) is controlled by variation in the length of a trinucleotide repeat motif 5' to the promoter of each gene. However, such a mechanism was not detected in preliminary observations on M. synoviae. Thus, the basis for control of variation in the vlhA gene family (which encodes the homologous haemagglutinin in M. synoviae) was investigated to enable comparison with its homologue in M. gallisepticum and with other lipoprotein gene families in mycoplasmas. The start point of transcription was identified 119 bp upstream of the initiation codon, but features associated with control of transcription in other mycoplasma lipoprotein genes were not seen. Comparison of three copies of vlhA revealed considerable sequence divergence at the 3' end of the gene, but conservation of the 5' end. Southern blot analysis of M. synoviae genomic DNA revealed that the promoter region and part of the conserved 5' coding sequence occurred as a single copy, whereas the remainder of the coding sequence occurred as multiple copies. A 9.7 kb fragment of the genome was found to contain eight tandemly repeated regions partially homologous to vlhA, all lacking the putative promoter region and the single-copy 5' end of vlhA, but extending over one of four distinct overlapping regions of the 3' coding sequence. Examination of sequential clones of M. synoviae established that unidirectional recombination occurs between the pseudogenes and the expressed vlhA, with duplication of pseudogene sequence and loss of the corresponding region previously seen in the expressed gene. Expression of the 5' end of two variants of the vlhA gene showed that they differed in their reaction with monoclonal antibodies specific for this region. These data suggest that the control of vlhA antigenic variation in M. synoviae is achieved by multiple gene conversion events using a repertoire of coding sequences to generate a chimeric expressed gene, with the greatest potential for variation generated in the region encoding the haemagglutinin. Thus, completely distinct mechanisms have been adopted to control antigenic variation in homologous gene families. PMID- 10692168 TI - Transfer RNA modification, temperature and DNA superhelicity have a common target in the regulatory network of the virulence of Shigella flexneri: the expression of the virF gene. AB - Full expression of the virulence genes of Shigella flexneri requires the presence of two modified nucleosides in the tRNA [queuosine, Q34, present in the wobble position (position 34) and 2-methylthio-N6-isopentenyladenosine (ms2i6A37, adjacent to and 3' of the anticodon)]. The synthesis of these two nucleosides depends on the products of the tgt and miaA genes respectively. We have shown that the intracellular concentration of the virulence-related transcriptional regulator VirF is reduced in the absence of either of these modified nucleosides. The intracellular concentration of VirF is correlated with the expression of the virulence genes. Overproduction of VirF in the tgt and the miaA mutants suppressed the less virulent (tgt) or the avirulent (miaA) phenotypes respectively, caused by the tRNA modification deficiency. This suggests that the primary result of undermodification of the tRNA is a poor translation of virF mRNA and not of any other mRNA whose product acts downstream of the action of VirF. Shigella showed no virulence phenotypes at 30 degrees C, but forced synthesis of VirF at 30 degrees C induced the virulence phenotype at this low temperature. In addition, removal of the known gene silencer H-NS by a mutation in its structural gene hns increased the synthesis of VirF at low temperature and thus induced a virulent phenotype at 30 degrees C. Conversely, decreased expression of VirF at 37 degrees C induced by the addition of novobiocin, a known inhibitor of gyrase, led to an avirulent phenotype. We conclude that tRNA modification, temperature and superhelicity have the same target - the expression of VirF - to influence the expression of the central regulatory gene virB and thereby the virulence of Shigella. These results further strengthen the suggestion that the concentration of VirF is the critical factor in the regulation of virulence in Shigella. In addition, they emphasize the role of the bacterial translational machinery in the regulation of the expression of virulence genes which appears here quantitatively as important as the well established regulation on the transcriptional level. PMID- 10692169 TI - The essential protein fap7 is involved in the oxidative stress response of Saccharomyces cerevisiae. AB - Pos9 (Skn7) is an important transcription factor that, together with Yap1, induces the expression of oxidative stress target genes in Saccharomyces cerevisiae. The activation of Pos9 upon an oxidative stress signal occurs post translationally. In a mutant screen for factors involved in the activation of a Pos9-dependent reporter gene upon oxidative stress, we identified the mutant fap7 1 (for factor activating Pos9). This point mutant failed to activate a Gal4-Pos9 hybrid transcription factor, assayed by hydrogen peroxide-induced GAL1-lacZ reporter gene activities. Additionally, the fap7-1 mutant strain was sensitive to oxidative stress and revealed slow growth on glucose compared with the wild type. The fap7-1 mutation also affected the induction of the Pos9 target gene TPX1 and of a synthetic promoter previously identified to be regulated in a Yap1- and Pos9 dependent manner. This lack of induction was specific as the fap7-1 mutant response to other stresses such as sodium chloride or co-application of both hydrogen peroxide and sodium chloride was not affected, as tested with the Pos9 independent expression pattern of a TPS2-lacZ reporter system. We identified the gene YDL166c to be allelic to the FAP7 gene and to be essential. Fluorescence microscopy of Fap7-GFP fusion proteins indicated a nuclear localization of the Fap7 protein. Our data suggest that Fap7 is a nuclear factor important for Pos9 dependent target gene transcription upon oxidative stress. PMID- 10692170 TI - The putative invasion protein chaperone SicA acts together with InvF to activate the expression of Salmonella typhimurium virulence genes. AB - SigD and SigE (Salmonella invasion gene) are proteins needed for optimal invasion of Salmonella typhimurium into eukaryotic cells in vitro. SigD is a secreted protein and SigE is a putative chaperone required for SigD stability and/or secretion. SigD is secreted by a type III secretion apparatus encoded within a pathogenicity island on the Salmonella chromosome known as Salmonella pathogenicity island 1 (SPI1). The expression of sigDE, which is not linked to SPI1, is co-ordinately regulated with the SPI1 genes and is dependent on the transcriptional regulators SirA, HilA and InvF. These three proteins alone are unable to activate transcription from the sigD promoter in Escherichia coli, therefore it is likely that other factors are needed for expression. A screen for genes required for the expression of a sigD-lacZYA reporter fusion found a mutant with a transposon insertion in spaS, an SPI1 gene which encodes a putative inner membrane component of the type III secretion system. The expression of a SPI1 operon encoding a putative chaperone (SicA) and several secreted proteins (Sips B, C, D and A) was also reduced in this mutant. The regulation defect of the spaS mutant was complemented by sicA and not by spaS. Because sicA is encoded immediately downstream of spaS, the mutation in spaS was likely to be polar on the expression of sicA. In addition, a sicA disruption mutant was as defective as an invF deletion mutant for the expression of sigD, sicA and sipC reporter fusions. The introduction of plasmids encoding invF and sicA into a non pathogenic E. coli K-12 strain stimulated the transcription of both a sicA- and a sigD-lacZYA promoter fusion. This result suggests that InvF and SicA are sufficient for the expression of these genes. This is the first demonstration of a positive regulatory role for a putative type III secretion system chaperone in the expression of virulence genes. PMID- 10692171 TI - Application of short tandem repeat of genomic DNA and mitochondrial DNA for identification of mixed-up tissue specimens. AB - Deoxyribonucleic acid (DNA) typing methods, short tandem repeat of genomic DNA and mitochondrial DNA with the use of polymerase chain reaction amplification, were applied to formalin-fixed, paraffin-embedded tissues submitted for diagnosis, to identify and sort out mixed-up tissue specimens. These techniques were found to be reliable, reproducible and specific for personal identification, and thus to eliminate the need for further examinations and to prevent unnecessary surgery. PMID- 10692172 TI - Immunohistochemical detection of cytokeratin and epithelial membrane antigen in leiomyosarcoma: a systematic study of 100 cases. AB - Although 'aberrant' expression of the epithelial markers, cytokeratin (CK) and epithelial membrane antigen (EMA), in leiomyosarcoma has been described previously, there has not been a study of this phenomenon with clinicopathological correlation in a large series of lesions at different anatomical sites. We investigated systematically the immunohistochemical reactivity for CK and EMA in 100 cases of leiomyosarcoma. CK and EMA were positive in 38% and 44% of the cases, respectively. Although staining was usually focal, extensive immunoreactivity was observed in 11% with CK and 6% with EMA. There was no correlation between immunoreactivity for CK and EMA in leiomyosarcomas and non-neoplastic smooth muscle at the same location. Immunoreactivity for CK and EMA was not correlated with the location, age, sex, histological grade, or histological features, except for more frequent EMA positivity in vascular and uterine tumors than in soft tissue cases. These results indicate that CK and/or EMA-positive leiomyosarcomas do not have distinctive clinicopathological features differing from those of negative cases. However, the considerable frequency of immunoreactivity for these epithelial markers in leiomyosarcoma, occasionally with diffuse and strong immunopositivity, should be recognized as a potentially serious diagnostic pitfall in the differential diagnosis of other malignant spindle cell neoplasms. PMID- 10692173 TI - Existence and distribution of melanocytes and HMB-45-positive cells in the human minor salivary glands. AB - The existence and distribution of melanocytes in the human minor salivary glands were investigated in a series of autopsy and biopsy materials. The cells with the following characteristics were regarded as melanocytes; spindle-shaped or dendritic cells with fine granular pigments: (i) stained brownish-black by hematoxylin-eosin stain, and black by Masson-Fontana's silver impregnation method; and (ii) disappeared after treatment with peroxide and potassium permanganate solution. In addition, the expression of antigen identified by anti HMB-45 antibody in serial sections with melanocytes was examined. Melanocytes were found in eight (1.8%) of 445 cases, and there was no relationship between the existence of melanocytes and significant diseases of the subjects. Various numbers of melanocytes were distributed in fibrous tissue around the interlobular ducts, intralobular ducts and acini, but were not in direct contact with the epithelia. Neither melanocytes nor melanin granules were found in the salivary gland epithelia. HMB-45-positive cells without intracytoplasmic fine granules were found solitarily or in small groups in periductal and periacinar fibrous tissues with or without slight infiltration of small mononuclear cells. PMID- 10692174 TI - Functional aspects of megamitochondria isolated from hydrazine- and ethanol treated rat livers. AB - It is essential to analyze functions of megamitochondria (MG) to elucidate the mechanism of the formation of MG induced under various pathological conditions. The MG fraction obtained by a routine isolation procedure for normal mitochondria always consists of a mixed population of mitochondria enlarged to various degrees and also normal-sized ones. The purpose of the present study is to answer the question of whether or not data obtained from the MG fraction consisting of such a heterogeneous population of mitochondria with respect to their sizes really reflect functions of MG. In the present study mitochondria were obtained from the livers of rats treated with a 1% hydrazine diet for 8 days and those given 32% ethanol in drinking water for up to 2 months using various isolation procedures. Results obtained are summarized as follows: (i) mitochondria enlarged to various degrees and normal-sized ones are sometimes connected with each other by a narrow stalk in the hepatocyte of hydrazine-treated animals, and such connections are maintained to some extent when mitochondria are isolated; and (ii) mitochondria obtained from experimental animals by a routine isolation procedure for mitochondria ((700-7000)gR2"') and those obtained by alternative isolation procedure yielding the heavy ((500-2000)gR2"') and light ((2000-7000)gR2"') fractions show some functional similarities: decreases in the content of cytochrome a + a3; decreases in oxygen consumptions and phosphorylating abilities; decreases in monoamine oxidase and cytochrome c oxidase activities; lowered membrane potential of mitochondria; decreases in the rate of the generation of reactive oxygen species. These results may suggest that mitochondria enlarged to various degrees and normal-sized ones are functionally similar to each other and that the MG fraction obtained by a routine isolation procedure for normal mitochondria can be applied to the study of the function of MG. PMID- 10692175 TI - Endotoxin priming and liver damage by experimental duodenal obstruction in the rat. AB - To verify whether endotoxin (LPS) might act as a priming cofactor of liver injury caused by obstructing the duodenum, four groups of male Wistar rats were studied. The first two groups comprised rats in which a closed duodenal loop (CDL) was created: CDL, n = 6 and CDL + LPS, n = 7; the next two groups comprised sham operated animals: Sham n = 6 and Sham + LPS, n = 6. LPS, 400 microg/kg bodyweight, was administered i.p. to the rats belonging to groups CDL + LPS and Sham + LPS, 24 h before laparotomy. Twenty-four hours after laparotomy the animals were killed. Damage to bile ducts, extent and grading of coagulative and lytic spotty necrosis in liver tissue were evaluated morphologically. Coagulative necrosis was severe in four of seven rats of the group CDL + LPS, mild in six of six rats of group CDL, and absent in four of six and five of six rats of groups Sham and Sham + LPS (chi2 32.8, P = 0.0001). The animals of group CDL + LPS had more frequently diffuse lytic spotty necrosis than the animals in the three other groups (chi2 9.57 P<0.01). The results of our study indicate that, in rodents subjected to a closed duodenal loop, priming with LPS exacerbates liver injury due to cholate stasis. PMID- 10692176 TI - Primary hepatic leiomyosarcoma in a patient with hepatitis C virus-related liver cirrhosis. AB - We describe an autopsy case of primary hepatic leiomyosarcoma in a 68-year-old man with hepatitis C virus-related liver cirrhosis. The patient, who had a history of acute hepatitis 20 years previously, died of a ruptured hepatic tumor. At autopsy, a well-circumscribed 14 x 16 x 15 cm tumor replaced the medial site of the right hepatic lobe with multiple intrahepatic and distant metastases. Histologically the tumor, which had extensive central necrosis, consisted predominantly of well or moderately differentiated spindle-shaped cells, which were positive for smooth muscle actin and vimentin on immunohistochemical staining. In addition, clusters of markedly atypical cells and myxoid change of the matrix were discretely found in the focal and small areas of the tumor. These findings indicated that many sections were necessary for the histologically accurate estimation of primary hepatic smooth muscle tumor. The histological examination of a non-tumorous lesion showed liver cirrhosis. Hepatitis C virus was detected in the cytoplasm of cirrhotic hepatocytes by immunohistochemistry and reverse transcriptase-polymerase chain reaction, but not in the tumor cells. This suggested that the virus was not directly involved in the development of primary hepatic leiomyosarcoma. PMID- 10692177 TI - Adenocarcinoma of the renal pelvis with vimentin-positive intracytoplasmic inclusions. AB - A case of mucus-producing papillary adenocarcinoma of the renal pelvis associated with multiple renal calculi in a 16-year-old male is reported. The majority of the tumor cells contained large, round or cone-shaped, vimentin-positive intracytoplasmic inclusions, which exhibited some morphological and immunohistochemical resemblance to inclusions found in malignant rhabdoid tumor of the kidney. An intracytoplasmic accumulation of mucus was also noted in some tumor cells. The patient is free from recurrence and metastasis after a nephrectomy. PMID- 10692178 TI - Micropapillary variant of transitional cell carcinoma of the ureter. AB - Micropapillary variant of transitional cell carcinoma (TCC) is a rare entity, having a distinct micropapillary component mimicking papillary serous carcinoma of the ovary and has been reported exclusively in the urinary bladder. We experienced a case of micropapillary variant of TCC in the ureter. The tumor showed a predominant proportion of micropapillary component and accompanied a TCC in situ lesion and a high-grade TCC. A metastatic lesion in the regional lymph node also showed an entirely micropapillary pattern. Initially, our case was confused with adenocarcinoma, especially metastatic, because the micropapillary architecture resembled an abortive glandular structure and tumor cell nests were predominantly located in empty spaces mimicking vascular invasion. The patient died with peritoneal metastases 20 months after the initial diagnosis. We report the first case of a micropapillary variant of TCC occurring in the ureter. PMID- 10692179 TI - Anaplastic carcinoma of the pancreas with rhabdoid features. AB - The malignant rhabdoid tumor (MRT) is histologically characterized by the invasive proliferation of polygonal to ovoid cells with abundant eosinophilic cytoplasm and eccentric vesicular nuclei with a prominent nucleolus. MRT frequently occurs in the kidney, but may also arise in other organs. However, MRT should be strictly distinguished from carcinomas with rhabdoid features. A post mortem examination of a 68-year-old woman found an anaplastic carcinoma of the pancreas with rhabdoid features displaying extensive invasion into the neighboring tissues. To the best of our knowledge, this is the first case of a pancreatic tumor with rhabdoid features. Pathologists should consider that carcinomas showing rhabdoid features may also appear in the pancreas. As pancreatic tumors with rhabdoid features have characteristic histopathological features and poor prognosis compared to other pancreatic tumors, careful histopathological differential diagnosis is important. PMID- 10692180 TI - Sebaceous carcinoma of the breast. AB - We report on a rare distinctive variant of infiltrating ductal carcinoma characterized by sebaceous differentiation of tumor cells. The neoplasm was identified in a lumpectomy specimen from a 45-year-old woman with extensive metastatic disease. In addition to conventional in situ and invasive ductal components, approximately half of the tumor cells exhibited a phenotype resembling tumors of the sebaceous skin appendage with coarsely vacuolated cytoplasm and peripherally displaced nuclei. The sebaceous moiety was also present in the distant metastatic deposits. There was no evidence of mucin production by tumor cells. Ultrastructurally, empty-appearing non-membrane bound vacuoles attested to the sebaceous cells' lipid content. The immunoprofile of the lesion included positivity for cytokeratin and epithelial membrane antigen. Vimentin, S100 protein and carcinoembryonic antigen were not expressed. Most tumor cell nuclei reacted with antibodies to oestrogen and progesterone receptors but failed to show overexpression of the HER2/neu protein. The MIB-1 labeling index averaged 16%. At variance with sebaceous breast carcinomas on record, the present case is notable for its prolonged clinical course. PMID- 10692181 TI - Signet-ring cell carcinoma of the breast. AB - Primary signet-ring cell carcinoma of the breast is a very rare tumor and is not recognized as an independent entity of the World Health Organization classification of breast tumor. Primary signet-ring cell carcinoma of the breast is usually considered as a variant of mucinous carcinoma or lobular carcinoma and usually originates from the lobular epithelium. A case of primary signet-ring cell carcinoma of the breast in a 68-year-old woman is presented. Histologically, the majority of neoplastic cells had an intracytoplasmic mucin collection. The histological presence of ductal carcinoma in situ, absence of lobular lesion and immunoreactivity for estrogen and progesterone receptors implicated the tumor cells arising from ductal epithelium. The papillary or organoid growth pattern is characteristic in this case. The patient underwent a modified radical mastectomy and was subsequently followed up for 6 months. PMID- 10692182 TI - Blastomatous tumor with teratoid features of nasal cavity: report of a case and review of the literature. AB - A case of blastomatous tumor with teratoid features is presented. The polypoid mass was observed in the left nasal cavity of a 72-year-old man. Histologically, the lesion was composed of neuroepithelial cells with blastomatous appearance, cystic squamous nests filled with keratin materials, many mucous glands, complex tubular and glandular structures with edematous fibroblastic stroma. Sinonasal neoplasms including teratoid components and immature neuroepithelium are exceedingly rare. We suggest that the term 'immature teratoma' is more suitable than blastoma or blastomatous tumor when there is no carcinomatous or sarcomatous component besides the immature neuroepithelium and teratoid elements. PMID- 10692183 TI - Uterus-like mass of the small bowel mesentery. AB - A case of a uterus-like mass arising from the mesentery is reported. A mass measuring 14x11 cm was noted in the small bowel mesentery of a 59-year-old woman. Histologically, the lesion consisted of endometrial-type and fallopian tube-type mucosa surrounded by thick bundles of smooth muscle cells. Since the first report by Cozzutto in 1981, 10 cases of uterus-like mass, that included seven ovarian and three extraovarian cases, have been reported. To our knowledge, the present lesion was the first case originating from the mesenteric region. Three hypotheses of this rare lesion: (i) congenital anomaly; (ii) metaplasia; and (iii) heterotopia theories are reviewed. PMID- 10692185 TI - We Are Marching, Marching ellipsis. PMID- 10692184 TI - Proliferative activity and subtyping of calcifying odontogenic cyst. PMID- 10692186 TI - Prediction of short- and long-term outcomes by troponin T levels in low-risk patients evaluated for acute coronary syndromes. AB - STUDY OBJECTIVE: Recent reports suggest a short series of cardiac troponin (cTnT) testing effectively identifies patients at risk for cardiac events. However, there are few studies validating this strategy. The purpose of this study was to determine the ability of cTnT levels to predict short- and long-term outcomes in low-risk patients with suspected acute coronary syndromes. METHODS: This prospective longitudinal study was conducted in a 20-bed emergency department observation unit. Patients at low risk for acute coronary ischemia, with a normal creatine kinase-isoenzyme subunit MB (CKMB) index, were admitted to an observation unit for chest pain evaluation. Serum cTnT levels were measured at baseline and at 4, 8, and 16 hours after admission. The main outcome measures were adverse cardiac events (death, acute myocardial infarction, unstable angina, revascularization) during the index visit and within 6 months after discharge. Using manufacturer's recommendations, the cTnT level was considered abnormal if it exceeded 0.2 microg/L. RESULTS: Two hundred sixty-six patients were evaluated. Twenty-one (7.9%) had an adverse event during their index hospitalization. Troponin testing identified only 2 (9.5%) of these patients. Twenty (7.5%) had a cardiac event within 6 months; none were identified by cTnT testing. The sensitivity and specificity were 9.5% and 99.2%, respectively, at the index visit, and 0% and 98.4% at 6 months. The positive and negative predictive values were 50% and 93%, respectively, at the index visit; and 0% and 92% at 6 months. CONCLUSION: Determination of troponin T levels has a low sensitivity and high specificity for predicting outcomes in low-risk patients evaluated for suspected acute coronary syndromes. This study does not support a strategy of relying solely on troponin testing for disposition decisions. PMID- 10692187 TI - Emergency airway management in penetrating neck injury. AB - STUDY OBJECTIVES: Airway management in the context of penetrating neck injury is a challenging scenario. Management decisionmaking has not been well studied and the initial airway approach remains controversial. We examined various initial emergency airway techniques and their success in the setting of penetrating neck trauma. METHODS: A retrospective study was conducted of emergency department intubations in penetrating neck injury from January 1, 1993, to December 31, 1996, at a Level I trauma center. Cases of out-of-hospital traumatic arrest or out-of-hospital intubation were excluded. Successful airway management was defined as endotracheal tube placement confirmed by clinical evaluation, pulse oximetry, chest radiography, and end-tidal CO(2) detection. RESULTS: During the study period, 748 consecutive patients with penetrating neck injury were evaluated in the ED. Of these, 82 (11%) were deemed to require immediate airway management. Twenty-four of the 82 were excluded because of out-of-hospital traumatic arrest or out-of-hospital intubation, resulting in a study population of 58 patients. Of these 58 patients, 39 had initial rapid sequence intubation using succinylcholine with a 100% success rate. Five comatose patients had successful orotracheal intubation without paralysis, and 2 patients underwent successful emergency tracheostomy. The remaining 12 patients had initial fiberoptic intubation by otolaryngology clinicians, which was unsuccessful in 3 patients. All 3 of these patients were subsequently successfully orotracheally intubated using the rapid sequence intubation technique. Therefore, oral endotracheal intubation was the definitive method of airway management in 47 (81%) of the 58 patients and was successful in all cases. CONCLUSION: Rapid sequence intubation was the most commonly performed initial technique by emergency physicians and was safe and effective in all cases attempted. Furthermore, rapid sequence intubation methodology resulted in successful intubation of the fiberoptic intubation failures. Physicians with airway expertise should consider using rapid sequence intubation as an initial airway technique in managing patients with penetrating neck injury who require airway control. PMID- 10692188 TI - Use of alternative therapies among emergency department patients. AB - STUDY OBJECTIVE: To assess emergency department patients' use of alternative therapies. METHODS: This study used a cross-sectional observational survey of a convenience sample of ED patients. A trained research assistant administered a written questionnaire asking patients about alternative therapies. RESULTS: Of the 139 patients surveyed, 78 (56%) had tried alternative therapies in the past, 68 (87%) of whom believed that they were effective. The most frequently tried alternative therapies were massage therapy (31%), chiropractic (30%), herbs (24%), meditation (19%), and acupuncture (15%). Most patients (70%) who tried alternative therapies did not inform their physicians of such practice. CONCLUSION: Most ED patients in our sample had tried alternative therapies and among these patients, most did not inform their physicians. Herbal therapy in particular had been tried by about 1 in 4 patients. Emergency physicians should routinely question their patients regarding the use of alternative therapies, particularly herbal preparations, which may cause adverse effects. PMID- 10692189 TI - Does adjunctive midazolam reduce recovery agitation after ketamine sedation for pediatric procedures? A randomized, double-blind, placebo-controlled trial. AB - STUDY OBJECTIVE: Despite widespread use of adjunctive benzodiazepines during ketamine sedation, their efficacy in reducing recovery agitation in children has never been studied. We wished to characterize the nature and severity of recovery agitation after ketamine sedation in children treated in the emergency department and to determine whether the addition of adjunctive midazolam reduces the magnitude of such recovery agitation. METHODS: The study was a randomized, double blind, clinical trial of adjunctive midazolam versus placebo during ketamine sedation. We enrolled 104 children aged 12 months to 15 years (median age, 6 years) at a combined university medical center and children's hospital. Subjects received either intravenous midazolam (0.05 mg/kg up to 2 mg) or placebo after intravenous administration of a ketamine loading dose (1.5 mg/kg). Treating physicians and nurses independently noted the presence of crying, hallucinations, and nightmares during recovery and graded recovery agitation by using a 100-mm visual analog scale. Preprocedure agitation and external stimulation during recovery were also graded. The time from ketamine injection until each subject met the recovery criteria was recorded. RESULTS: Fifty-three subjects received midazolam, and 51 received placebo. Potentially confounding variables were similar between the groups. Sedation efficacy, adverse effects, and recovery time were also similar between groups. Interobserver agreement between physician and nurse assessments was substantial. Median physician assessment of recovery agitation was 4 mm (interquartile range, 2 to 19) in the midazolam group and 5 mm (interquartile range, 3 to 14) in the placebo group (difference -1; 95% confidence interval -3 to 2; P =.705). Recovery agitation was moderately correlated with preprocedure agitation (rho=0.486) but not with external stimulation during recovery (rho=0.147). CONCLUSION: Recovery agitation is common but generally of very low magnitude after ketamine sedation in children in the ED. We observed a median physician rating of 5 mm on a 100-mm visual analog scale, a score that we believe to be clinically insignificant. The degree of recovery agitation after ketamine sedation is significantly related to the degree of preprocedure agitation. In this study, concurrent midazolam did not diminish such agitation and had no measurably beneficial effect. Use of adjunctive benzodiazepines in pediatric ketamine sedation appears unnecessary. PMID- 10692190 TI - Prospective evaluation of mild to moderate pediatric acetaminophen exposures. AB - STUDY OBJECTIVE: To determine whether pediatric patients with acute, mild to moderate acetaminophen exposures, treated with home monitoring alone, develop systemic signs of hepatic injury. METHODS: A prospective, observational study of calls to a regional poison center over a 25-month period was performed. Patients were eligible for the study if they were younger than 7 years and had an acute maximum possible acetaminophen exposure of up to 200 mg/kg. Exclusion criteria included previous decontamination measures, possibility of ingestion of an extended-release preparation, health or medication issues that could increase susceptibility to hepatotoxicity, current symptoms of hepatotoxicity, and indeterminable ingestions. Study protocol included reviewing the signs and symptoms of early and late acetaminophen toxicity, a 4- to 6-hour follow-up call, and a 72-hour follow-up call. Outcome measures were defined as a verbal report by the patient's parent or guardian of the presence or absence of signs or symptoms of hepatotoxicity. RESULTS: A total of 1,039 patients were enrolled in the study, including 519 girls and 520 boys, with exposures ranging from 20 to 200 mg/kg. Eighteen patients were lost to follow-up; data were incomplete for 2 patients. At 72-hour follow-up, the remaining 1,019 patients were all doing well, without signs or symptoms of hepatotoxicity. CONCLUSION: On the basis of these data, pediatric patients with acute acetaminophen exposures of up to 200 mg/kg, treated with home monitoring alone, do not develop signs or symptoms of hepatic injury. PMID- 10692191 TI - Injury prevention and emergency medical services for children in a managed care environment. AB - Each year, 1 in 5 US children receives medical care as a result of injury. Injuries are the leading cause of medical spending for children ages 5 to 21 years, accounting for more than 20% of hospital admissions and days spent in the hospital. Pediatric injuries become an important issue for managed care organizations because of concern for member safety and increasing medical costs related to treatment. Because effective prevention decreases health care consumption, injury prevention often costs less than treating injuries. Simple devices, such as bicycle helmets, smoke detectors, and child safety seats, help keep children safe and save money. Appropriate emergency care at the scene of an injury, poison control centers that dispense expert advice over the telephone, and triaged regional trauma systems improve the outcome and save money at the same time. This article continues the white paper series by the Emergency Medical Services for Children Managed Care Task Force. PMID- 10692192 TI - Adolescent injury in the emergency department: opportunity for alcohol interventions? AB - STUDY OBJECTIVE: Alcohol, the most commonly used substance among adolescents, is frequently associated with injury. Little is known regarding the drinking characteristics of injured adolescents. Such data are critical for developing emergency department interventions to decrease alcohol-related injury among adolescents. We sought to describe the drinking characteristics of injured adolescents and to describe the relationship of injury severity and mechanisms with drinking characteristics. METHODS: This study was a prospective cohort study performed in a university hospital (sampled May 1, 1995, to July 15, 1995) and a large urban teaching hospital (sampled May 1, 1996, to August 1, 1996). The participants were aged 12 to 20 years, presenting within 6 hours of an injury. We performed a saliva alcohol test and self-administered questionnaire. Age, sex, E code, injury severity score (ISS), and ED disposition were recorded. An alcohol frequency/quantity index was calculated. Descriptive statistics and 95% confidence intervals were calculated. RESULTS: Two hundred sixty-three patients with a mean age of 17 years and a mean ISS of 2.1 (SD 3.5) were recruited. One hundred fifty-two (50%) were males, and 33 (13%) were admitted. Ten (4%) patients had a positive saliva alcohol test response. On average, within the last year, these adolescents had 1.7 adverse alcohol consequences. Sixty percent drank in unsupervised settings, and 36% reported drinking 5 or more drinks in a row. CONCLUSION: Alcohol use/misuse is a substantial problem among injured adolescents regardless of severity or mechanism of injury. ED physicians should consider screening/intervention or primary prevention of alcohol problems for all injured adolescents. PMID- 10692193 TI - Lethality of firearm-related injuries in the United States population. AB - STUDY OBJECTIVE: To characterize differences in the lethality of firearm-related injuries in selected demographic subgroups using national representative data on fatal and nonfatal firearm-related injuries. We also characterize the lethality of firearm-related injuries by intent of injury and anatomic location of the gunshot wound. METHODS: We analyzed case-fatality rates (CFRs) of firearm-related injuries in the United States by using death data from the National Vital Statistics System and data on nonfatal injuries treated in US hospital emergency departments from the National Electronic Injury Surveillance System. National estimates of crude and age-adjusted CFRs are presented by sex, race/ethnicity, age, intent, and primary body part affected. RESULTS: Each year during the study period (July 1992 through December 1995), an estimated 132,687 persons sustained gunshot wounds that resulted in death or treatment in an ED. The overall age adjusted CFR among persons who sustained firearm-related injuries was 31.7% (95% confidence interval [CI] 27.7 to 35.6). The age-adjusted CFR for persons who were alive when they arrived for treatment in an ED (11. 3%; 95% CI 9.4 to 13.2) was about one third as large as the overall CFR. The age-adjusted CFR varied by sex, race/ethnicity, and age, but these differences depended on intent of injury. For assaultive injuries, the age-adjusted CFR was 1.4 times higher for females (28. 7%) than males (20.6%). For intentionally self-inflicted injuries, the age adjusted CFR was 1.1 higher for males (77.7%) than females (69.1%). For assaults, the age-adjusted CFR was 1.5 times higher for whites (29.5%) than blacks (19.2%). For assaultive and intentionally self-inflicted injuries among persons 15 years and older, the age-specific CFR increased with age. Persons shot in the head (age adjusted CFR, 61.0%) were 3.3 times as likely to die as those shot in other body parts (age-adjusted CFR, 18.7%). CONCLUSION: The lethality of firearm-related injuries was influenced strongly by the intent of injury and body part affected. The high lethality of firearm-related injuries relative to other major causes of injury emphasizes the need to continue prevention efforts and efforts to improve access to care and treatment (including emergency medical and acute care services) to reduce the number and increase survivability of firearm-related injuries. PMID- 10692194 TI - Effect of state legislation prohibiting denial of emergency department patient claims. AB - STUDY OBJECTIVE: On July 1, 1996, two Florida state laws were implemented to prevent denial of legitimate patient claims. Our objective was to determine whether the laws have been effective in reducing inappropriate denials as measured by the proportion of claims and charges denied. METHODS: A comprehensive set of claims for in-state emergency physician services from a physician billing company were analyzed for the period January 1996 through June 1997, covering 6 months before and 12 months after the effective date of state legislation. The number of facilities included in the data varied from 55 to 67 (mean 63). Denials were classified into 6 categories by payer type. Gross denials were those claims that were completely not paid by the payer, net denials represented the amount denied after patient payments. Downcoding was not examined in this study. Main outcome measures were the proportion of claims and charges denied before and after July 1, 1996. RESULTS: The classification of relative proportions of primary payers did not change appreciably over the study period. The proportion of denied claims decreased significantly (Kruskal-Wallis P <.001), starting 2 months after implementation. CONCLUSION: After initiation of state legislation, payers continue to inappropriately deny claims, although the number of claims and total charges denied has decreased. In response to this legislation, payers are denying larger claims and patient copayments have increased. PMID- 10692195 TI - Managed care organization authorization denials: lack of patient knowledge and timely alternative ambulatory care. AB - STUDY OBJECTIVE: To assess patient knowledge of managed care organization (MCO) regulations, availability of alternative ambulatory care, and patient outcome after MCO insurance authorization denial for an emergency department visit. METHODS: A medical screening examination and a follow-up structured interview were conducted with patients denied authorization for ED visits. The study was conducted at a large urban hospital with 36,000 annual ED visits and 40% MCO patients. RESULTS: During a 7-month period, 151 patients did not receive MCO authorization for ED care. The interview response rate was 75% (104/138) with 13 patients excluded. Eighty-three percent (86/104) of respondents came to the ED because they believed their problem was an emergency. Four percent (4/104) of the respondents had been instructed to go to the ED but were later denied authorization, whereas 85.6% (89/104) did not know that the MCO could deny payment. Only 37% (38/104) of the respondents reported having received instruction on the MCO preauthorization process, whereas of the 19% who contacted their MCO as instructed, all resulted in scheduling difficulties. Although 57% (59/104) received follow-up within 24 hours, 11% (11/104) of the respondents had a subsequent return visit to the ED with a subsequent admission rate of 4% (4/104). CONCLUSION: Few patients are aware of the need for MCO preauthorization for ED care, and almost half do not receive alternative care within 24 hours. A significant number of patients (11%) returned to the ED with an admission rate of 4%. PMID- 10692196 TI - Inadequate hospital reimbursement for victims of motor vehicle crashes due to health reform legislation. AB - STUDY OBJECTIVE: Effective for 1997, health reform legislation in New York resulted in a change in hospital reimbursement for victims of motor vehicle crashes. We evaluated the impact of this change from no-fault to Medicaid rates on the financial viability of a regional trauma center within an academic medical center. METHODS: This study represents a retrospective review of the trauma registry for all motor vehicle-related injuries (meeting the statewide definition of trauma) admitted to a regional trauma center for a 9-month period just before the legislation implementation date. Charges, costs, and projected reimbursement were calculated by standard hospital accounting methods. Profit or loss (reimbursement minus costs) was calculated by standard hospital accounting methods for each admission using no-fault and Medicaid reimbursement rates. RESULTS: One hundred seventy-three cases during the 9-month period generated total charges of $4,112,174, total costs of $3,447,110, and estimated total profit of $800,084 ($4,625 per case) using no-fault reimbursement and a total loss of $184,154 ($1,064 per case) using Medicaid reimbursement. For the 31 patients with diagnosis-related groups (DRGs) that were specifically created in New York to ensure adequate reimbursement for multiple significant trauma (730 through 734 and 792 through 794), no-fault reimbursement resulted in an average profit of $371 per case and Medicaid generated a loss of $6,118 per case. Actual payments for the study population were almost $500,000 less than estimated. CONCLUSION: Changes in rates of no-fault insurance payments to hospitals will result in inadequate reimbursement for motor vehicle crash victims admitted to a regional trauma center, undermining the viability of the regional trauma system. PMID- 10692197 TI - Effect of a state definition of an "emergency medical condition" legislation on medicaid managed care organization reimbursement. AB - STUDY OBJECTIVE: The state of Michigan passed Public Act 136 of 1997 requiring Medicaid managed care organizations (MMCOs) to pay for emergency services whenever presenting symptoms constituted an "emergency medical condition." The objective of this study was to evaluate MMCO reimbursement before and after enactment of this state law. METHODS: We conducted a retrospective comparison of reimbursement for lacerations needing repair (identified using Current Procedural Terminology codes from computerized billing data) for 2 time periods (before the state law was applicable [January through March 1998] and after the state law was applicable [April through June 1998]) from MMCO enrollees in 7 different MMCOs presenting to 4 urban emergency departments. Three months after billing submission was allowed for payment. Only refusal of reimbursement was evaluated. Data were analyzed using chi(2) and Fisher's exact test (values of P <.05 were considered significant). RESULTS: The total number of MMCO patients evaluated/total number of ED patients evaluated for the 2 periods was 1,769/32,646 and 3, 376/30,901, respectively (P <.05). The number of MMCO lacerations with no reimbursement/total number of MMCO lacerations for the 2 periods was 4/135 (3%) and 78/196 (40%), respectively (P <.001). CONCLUSION: Reimbursement by MMCOs for a procedure chosen to reflect a state-defined "emergency medical condition" is inadequate and significantly decreased during the 2 periods, with a significant increase in MMCO patients evaluated. PMID- 10692198 TI - Reimbursement impact of medicaid managed care organizations replacing standard medicaid. AB - STUDY OBJECTIVE: To evaluate the reimbursement difference for Medicaid managed care organization (MMCO) enrollees compared with Medicaid enrollees for emergency department patients with disease conditions that appear to meet the "prudent layperson" definition of an emergency medical condition. METHODS: This study used a retrospective reimbursement review of computerized billing data of reimbursement denials for 4 procedures (using Current Procedural Terminology codes for endotracheal intubation, cardiopulmonary resuscitation, central line placement, and lumbar puncture) and 1 International Classification of Diseases, ninth revision condition (chest pain) on MMCO patients from 7 MMCOs compared with standard Medicaid patients presenting to 4 EDs during a 6-month period (January through June 1998). Exclusion criteria were late bills that did not allow at least 90 days for payment and bills submitted on behalf of patients that were not covered at the time of service by Medicaid or MMCO. Data were analyzed using Fisher's exact test. RESULTS: The total number of MMCO and Medicaid patients evaluated/total ED patients evaluated was 5,153/63,552 and 6,020/63, 552, respectively. The number of nonreimbursed procedures/total number of procedures performed on MMCO and Medicaid patients was 35/93 and 14/88, respectively (P <.05). The number of nonreimbursed chest pain patients/total chest pain patients evaluated for MMCO and Medicaid enrollees was 65/277 and 12/199, respectively (P <.05). CONCLUSION: MMCOs reimburse significantly less than Medicaid does for ED patients with conditions that a prudent layperson would consider an emergency. PMID- 10692199 TI - Analysis of insurance payment denials using the prudent layperson standard. AB - STUDY OBJECTIVES: To review a sample of emergency department payment denials characterized as "not a medical emergency" and to determine medical necessity for each visit using an arbitrary "prudent layperson" standard. METHODS: This study was conducted at a university hospital and was an analysis of a convenience sample of ED payment denials classified as "not a medical emergency" by 2 managed care providers. Each corresponding visit was analyzed if the bill was still outstanding in September 1998. ED records were analyzed for chief complaint and risk factors for morbidity. Any minor disorder lasting 1 day or more and with normal vital signs recorded was considered to not meet the prudent layperson standard of an emergency. Visits for minor trauma that occurred the same day that also required radiographs or suturing were considered emergencies. RESULTS: Two hundred ED visits were retrospectively reviewed. Payer 1 denied 44 visits, of which 38 (86%) met the prudent layperson standard; payer 2 denied 156 visits, of which 113 (62%) met the standard (P >.05). CONCLUSION: A large proportion of ED visits for which payment is denied as "not a medical emergency" may meet the prudent layperson definition of an emergency. PMID- 10692200 TI - The uninsured: emergency medicine's challenge to our political leaders. PMID- 10692201 TI - Midazolam with ketamine: who benefits? PMID- 10692202 TI - Cerebral arterial gas embolism by helium: an unusual case successfully treated with hyperbaric oxygen and lidocaine. AB - A 27-year-old man inhaled helium from an unregulated pressurized cylinder and underwent cerebral arterial gas embolism (CAGE), leaving him blind and with radiologic evidence initially suggesting cortical infarction. There was complete recovery of vision and substantial regression of the radiologic changes after 4 hyperbaric oxygen treatments and a 54-hour lidocaine infusion, which began 6 hours after the accident. This is the second reported case of CAGE occurring by this mechanism and the first case of unequivocal CAGE in which lidocaine has been used as an adjunctive treatment with hyperbaric oxygen. PMID- 10692203 TI - Controversial company: the prudent layperson standard and the patients' bill of rights. PMID- 10692204 TI - Use of pulsed-field gel electrophoresis for investigation of a cluster of invasive group A streptococcal illness - Spokane, Washington, 1999. PMID- 10692205 TI - Mooki and Ben. PMID- 10692206 TI - Methanol intoxication. PMID- 10692207 TI - Potential for duragesic patch abuse. PMID- 10692208 TI - Disaster medical education for all physicians and physician extenders. PMID- 10692209 TI - Disaster Medical Education for All Physicians and Physician Extenders. PMID- 10692211 TI - Esophageal cytology in the follow-up of patients with treated upper aerodigestive tract malignancies. AB - BACKGROUND: Patients with an history of carcinoma of the upper aerodigestive tract are at high risk for recurrence or the development of new tumors in this region. In the majority of follow-up protocols, these patients undergo radiologic and endoscopic evaluation as a means of surveillance for the early detection of recurrence. The brush biopsy-capsule technique represents a noninvasive and inexpensive screening device for this patient population. In the current study, the authors retrospectively assessed the sensitivity, specificity, and predictive value of esophageal brush-capsule cytology for the detection of malignant lesions of the upper aerodigestive tract in this high risk patient population. METHODS: Cytologic specimens from 334 patients with previously treated upper aerodigestive malignancies were available for review. The cytologic, endoscopic, and clinical follow-up of each case were studied over a follow-up period of 3 years. Gold standard was the clinical follow-up for the negative cases (who were not submitted to biopsy) and biopsy for the positive cases. Sensitivity, specificity, and predictive value were calculated. RESULTS: Using cytology 33 malignancies were detected in 25 patients during a 3-year follow-up period. The test was found to have a sensitivity of 88.7% and a specificity of 90.7%. In 66% of cases the malignancies were located in the oropharynx; the others were located in the esophagus. In 70% of cases the malignancies were detected at an early stage. CONCLUSIONS: Esophageal brush-capsule cytology is a simple noninvasive technique that has been proven to be useful in the early detection of metachronous and recurrent neoplasms in the follow-up of patients with previously treated carcinomas of the ear, nose, and throat. PMID- 10692210 TI - Molecular biology in cytopathology: current applications and future directions. AB - The use of molecular techniques in cytopathology has become an accepted and billable practice for certain applications. Other applications still are in the developmental or experimental stages but may soon be clinically useful. The current study provides a review of these techniques including molecular detection of clonality, in situ hybridization, loss of heterozygosity, and single base mutation detection. A number of new molecular techniques recently have been described that may have a dramatic impact on diagnostic pathology. These techniques, in situ detection of single base mutations and microarray technology, are introduced and discussed in the context of potential applications to cytopathology in the future. PMID- 10692212 TI - Epithelial cells and other cytologic features of pseudomyxoma peritonei in patients with ovarian and/or appendiceal mucinous neoplasms: a study of 12 patients including 5 men. AB - BACKGROUND: Pseudomyxoma peritonei (PP) is a rare condition. Cytologic evaluation of peritoneal fluid often is an initial diagnostic test for possible ovarian and/or appendiceal primary tumors. Previous studies suggest that patients with PP who have epithelial cells (ECs) in their peritoneal fluid usually have a less favorable prognosis than patients with acellular PP. To the authors' knowledge, few reports of PP in the cytologic literature cite the presence of ECs. METHODS: Twelve cases of PP diagnosed by cytologic examination at the University of Texas M. D. Anderson Cancer Center over 15 years were identified. In all cases, primary tumors were confirmed histologically. All available cytologic smears and cell block sections were reviewed for cytomorphology, with particular attention given to the presence of ECs. A correlation between the presence of ECs and patient outcome also was sought (median follow-up, 26 months). RESULTS: Two patients had ovarian neoplasms, six patients (one female and five males) had appendiceal neoplasms, and four patients had synchronous ovarian and appendiceal tumors. Cytologic features included mucin pools (12 of 12 patients), ECs (11 of 12 patients), mesothelial or mesothelial-like cells (10 of 12 patients), histiocytes (11 of 12 patients), and fibroblast-like or spindle cells (6 of 12 patients). ECs were columnar with mucinous features in the majority of cases, and the number of ECs in each case was variable, ranging from 1+ (rare) to 3+ (many). Of the 11 patients with available follow-up data, 6 had recurrent disease, 4 had persistent disease, and 1 patient with acellular PP was alive without clinical evidence of disease after 24 months of follow-up. CONCLUSIONS: Unlike previous PP cytology reports, the current study frequently identified ECs (92%). Because of the potential prognostic implication of ECs in patients with PP, a diligent search for ECs is warranted. Indication of the presence or absence of ECs in the cytology report may be useful when PP is diagnosed. PMID- 10692213 TI - Cytology of thymomas: emphasis on morphology and correlation with histologic subtypes. AB - BACKGROUND: Aspirates of thymomas are distinguishable from other lesions and fine needle aspiration (FNA) is a proven method for investigating mediastinal masses. METHODS: Thirty-four cytology specimens of thymomas from 31 patients were examined. Corresponding surgical materials were available in 32 cases. Ten cases were benign and 22 were malignant. Cytologic features of these thymomas were correlated with various histologic classification systems and with biologic behavior. Dual epithelial and lymphoid populations and irregular cohesive tissue fragments of varying proportions of lymphoid and epithelial cells were characteristic of all aspirates. RESULTS: Using the Lattes-Bernatz classification, 10 cases predominately were lymphocytic, 3 cases predominately were epithelial, 3 cases predominately were spindle, 15 cases predominately were mixed, and 1 case was a thymic carcinoma. In the Muller-Hermelink classification, 3 cases were medullary, 12 were mixed, 8 predominately were cortical, 2 were cortical, 6 were well differentiated thymic carcinoma, and 1 was a poorly differentiated thymic carcinoma. In the majority of the cases the epithelial cells were round to oval. Spindle cells and a mixture of round to oval and spindle cells also were observed. No cytologic feature was found to correlate significantly with any classification scheme. Necrosis was present in 5 of the 32 aspirates, most frequently in malignant tumors. Thymomas showing predominately spindle cells frequently were encapsulated. Tumors with predominantly round to oval cells or a mixed population behaved more aggressively than those with spindle cells. Tumors that were well encapsulated and benign clinically tended to possess benign-appearing nuclei. Among the 22 invasive or malignant lesions, 8 exhibited moderate to marked cytologic atypia and 14 showed little or no atypia. No atypia was observed in benign tumors. CONCLUSIONS: The presence of cytologic atypia of epithelial cells may be helpful in predicting aggressiveness. However, the absence of atypia and necrosis may not imply a benign course. Correlation with clinical and radiographic findings should be sought. PMID- 10692214 TI - Cytologic features of proliferative breast disease: a study designed to minimize sampling error. AB - BACKGROUND: Assessment of cytologic features that allow accurate classification of proliferative breast disease has been hampered by sampling errors when fine needle aspirations have been compared with their corresponding histologic sections. METHODS: To allow for optimal cytohistologic correlation, 2 smears (1 hematoxylin and eosin-stained and 1 Diff-Quik-stained) were prepared from each of 98 breast biopsies without mass lesions and compared with the corresponding histologic sections of the scraped area. Each smear was reviewed in a blinded fashion and assessed for cellularity, background elements, cytoarchitectural features of cell groups, and nuclear features by 2 reviewers. Smears were then classified as nonproliferative breast disease (NPBD), proliferative breast disease without atypia (PBD) or with atypia (PBDA), or DCIS, based on review of the corresponding histologic sections. RESULTS: When comparing NPBD/PBD (n = 86) with PBDA/DCIS (n = 12), smears from PBDA/DCIS were significantly (by the Fisher exact test or Wilcoxon rank sum P values with adjustment for multiple comparisons) more likely to be cellular; contain single cells and necrosis; exhibit nuclear overlap and cytoplasmic vacuoles; have large nuclei, macronucleoli, pleomorphism, clumped chromatin, and hyperchromasia; and were less likely to have complex cell groups, monolayers, swirling, cohesion, and myoepithelial cells in epithelial sheets and the smear background. When NPBD (n = 53) and PBD (n = 33) were similarly compared, smears from PBD were more likely to exhibit larger and more complex cell groups, but they were otherwise similar to smears from NPBD. CONCLUSIONS: There are many cytologic features that will allow a distinction of NPBD/PBD from PBDA/DCIS, but relatively few that can aid in separating NPBD from PBD. PMID- 10692215 TI - Intranuclear holes (cytoplasmic pseudoinclusions) in parathyroid neoplasms, or "holes happen". AB - BACKGROUND: A parathyroid adenoma demonstrating intranuclear holes on aspiration cytology prompted a review of parathyroid neoplasms to determine the frequency of this phenomenon. METHODS: Aspiration cytology slides from 30 parathyroid adenomas and 1 parathyroid carcinoma were reviewed. In addition, histologic slides from 136 parathyroid adenomas and 7 parathyroid carcinomas were reviewed. Twenty-two cases had both cytologic and histologic slides available for review. The presence and approximate frequency of intranuclear holes were recorded. RESULTS: On cytologic smears, intranuclear holes were found in 3 of the 31 cases reviewed (2 adenomas and 1 carcinoma). The holes were frequent in only one adenoma (the index case) and moderate-rare in the other two cases. Review of tissue slides showed holes in 9 of 136 adenomas and 3 of 7 carcinomas; 2 adenomas and 2 carcinomas had frequent holes, whereas the remainder had holes infrequently. CONCLUSIONS: Intranuclear holes do occur in parathyroid neoplasia, in rare cases with considerable frequency. When intranuclear holes are observed in aspiration cytology specimens from the thyroid region, various thyroid and parathyroid conditions must be considered. PMID- 10692216 TI - Fine-needle aspiration cytology of bone: accuracy and pitfalls of cytodiagnosis. AB - BACKGROUND: Fine-needle aspiration cytology has proved to be an accurate, cost effective, and safe technique for diagnosing inflammatory and neoplastic lesions at different body sites. Its applicability in bone pathology, however, has been controversial due to a high percentage of inadequate samples and nonspecific results in the diagnosis of primary bone lesions. In this study, the diagnostic accuracy of the technique and its capacity for diagnosing primary bone lesions were assessed. In addition, the authors analyzed the diagnostic limitations with focus on specimen adequacy. METHODS: The authors reviewed 314 consecutive fine needle aspirations of bone from 308 patients. Direct or cytospin smears from aspirated material were fixed in 95% alcohol and stained by a modified Papanicolaou technique. Ninety-seven smears (31%) initially were considered unsatisfactory and excluded from the study. A diagnosis was rendered in 217 cases (69%), which were classified into 4 categories: primary bone lesions (benign and malignant) (42%), metastatic bone tumors (37%), suspicious for malignancy (5%), and negative (16%). RESULTS: The overall accuracy was 95%. Seventy-eight percent of primary bone lesions were correctly diagnosed by cytology. All cases diagnosed as metastatic by cytology were correct. The authors encountered difficulties diagnosing fibro-osseous lesions. Thirteen percent of cases were erroneously diagnosed as "negative" or "inflammatory conditions." On review, the absence of adequate cytologic material was noted in all of them. This sampling error could have been avoided by the presence of an on-site cytopathologist. CONCLUSIONS: Fine-needle aspiration of bone is a simple, reliable, and accurate diagnostic technique that can facilitate patient management and preoperative decision-making and/or avoid unnecessary invasive procedures for patients with primary or metastatic bone lesions. However, the radiologist, cytopathologist, and orthopedic surgeon should work together for optimal results. Moreover, a definitive pathologic diagnosis should not be issued if diagnostic material is not adequate and/or clinicoradiologic information is incompatible. PMID- 10692217 TI - Cytologic differential diagnosis among reactive mesothelial cells, malignant mesothelioma, and adenocarcinoma: utility of combined E-cadherin and calretinin immunostaining. AB - BACKGROUND: The differential diagnosis between reactive mesothelial cells (RMs), malignant mesotheliomas (MMs), and adenocarcinomas (ACs) is often difficult in cytologic specimens, and the utility of various immunohistochemical markers have been explored. Because recent immunohistologic studies have suggested that E cadherin (E-cad) and calretinin (Cal) may be useful markers for epithelial and mesothelial differentiations, respectively, the authors investigated their utility in cytologic diagnosis. METHODS: In this retrospective study, immunostaining was performed on smears retrieved from Papanicolaou-stained slides of effusions using the labeled streptavidin-biotin method. Sixteen cases of RM, 9 cases of MM, and 52 cases of AC from various sites, including 13 pulmonary primaries, were examined with primary antibodies against E-cad and Cal. RESULTS: The positive rates for E-cad and Cal, respectively, were as follows: RM, 0/16 (0%) and 16/16 (100%); MM, 9/9 (100%) and 8/8 (100%); and AC, 45/52 (86.5%) and 0/51 (0%). The E-cad expression by neoplastic cells was strongest in the intercellular junctions, and poorly differentiated neoplastic cells in the single cell form showed the weakest expression. CONCLUSIONS: In contrast to the results of previous immunohistochemical studies, the current study indicates that MMs constantly express E-cad, whereas RMs lack its expression in cytologic specimens, which would be useful in the differential diagnosis between the two. On the other hand, E-cad expression is not reliable for distinguishing AC from MM. The Cal expression can be a very useful marker for the distinction between AC and the mesothelial lineage. The combined immunostaining for E-cad and Cal has utility in differential diagnosis among RM, MM, and AC. PMID- 10692218 TI - Comparison of the results of immunocytochemical assays for biologic variables on preoperative fine-needle aspirates and on surgical specimens of primary breast carcinomas. AB - BACKGROUND: Fine-needle aspiration biopsy (FNAB) is a well-documented procedure for the diagnosis and biologic characterization of breast carcinoma. In order to compare the immunocytochemical expression of biologic parameters on cytology and on histology, estrogen receptor (ER) and progesterone receptor (PgR) status, p53 protein expression, and Ki67 growth fraction were evaluated on presurgical fine needle aspirates (FNAs) from breast carcinoma patients and on the corresponding surgical samples prior to any systemic therapy. METHODS: FNAs were performed on 104 patients with primary breast carcinoma at the time of diagnosis and subjected to immunocytochemical evaluation of ER, PgR, p53, and Ki67. The same parameters were immunohistochemically evaluated on the corresponding paraffin embedded sections. RESULTS: ER, PgR, p53, and Ki67 were evaluable on FNAs and on paired tissue sections in 100, 97, 68, and 84 cases, respectively. Concordance between cytology and histology was 89% for ER, 78% for PgR, 79% for p53, and 70% for Ki67. CONCLUSIONS: The concordance between the results of immunocytochemical evaluation of ER, PgR, p53, and Ki67, on both cytology and histology, underscores the reliability of the biologic characterization of breast carcinoma by FNAB. This approach could be particularly useful in predicting prognosis and response to treatment in patients who are candidates for neoadjuvant chemotherapy and/or endocrine therapy. PMID- 10692219 TI - Utility of liquid-based cytology for cervical carcinoma screening. PMID- 10692220 TI - Author reply PMID- 10692221 TI - Multiplex polymerase chain reaction-based analysis of T-cell receptor gamma gene rearrangements for the determination of T-lymphocyte clonality. AB - Determination of the frequency of mutations at hprt or other loci in human lymphocytes provides a useful biomarker for human exposure to mutagens. One problem, however, is distinguishing between unique mutants and sibling mutants arising as progeny of an earlier mutant cell. We have developed a multiplex polymerase chain reaction (PCR)-based method to analyze T-cell receptor (TCR) gamma gene rearrangements for determination of T-cell clonality in mutational spectrum analysis. PCR primers for different subgroups of the V gene segment of the TCR gamma gene were selected at different sites in the TCR gamma gene so that the size of PCR products could define which V subgroup was involved in rearranged TCR gamma genes; gamma genes involving different V and J subgroups could be determined directly by PCR. Mutant T-lymphocytes with rearranged TCR gamma genes containing the same V and J subgroups were analyzed using PCR-based denaturing polyacrylamide gel electrophoresis. All of the 161 hprt mutant clones analyzed contained rearranged TCR gamma genes. Rearrangements among all subgroups of the V and J gene segments of the TCR gamma gene could be detected. VgammaI and Jgamma1/2 subgroups were involved in 69 and 71% of rearranged TCR gamma genes, respectively. This PCR-based analysis of TCR gamma gene rearrangements provides a simple and comprehensive method for identifying the clonality of mutant T lymphocytes in human hprt mutant lymphocyte assay and mutational spectrum analysis. PMID- 10692222 TI - Phthalates demonstrate genotoxicity on human mucosa of the upper aerodigestive tract. AB - Various phthalate compounds are used as softeners and plasticizers in a wide range of plastic materials. There has been a growing concern regarding a possible health hazard to humans. The mucosa of the upper aerodigestive tract is the organ of first contact for the majority of xenobiotics, such as phthalates, entering the body. Still, there is a lack of information concerning possible carcinogenicity of phthalates in the upper aerodigestive tract. This motivated us to investigate their genotoxic effects on human epithelia: human mucosal cells derived from biopsies harvested during surgery of the oropharynx and the inferior nasal turbinate, respectively. The alkaline version of the microgel electrophoresis assay was used to detect single-strand breaks in the DNA following incubation with dibutylphthalate (DBP) and diisobutylphthalate (DiBP). DNA damage was induced by both DBP and DiBP in oropharyngeal and nasal mucosa, though the effect of DiBP was more pronounced than that of DBP. Nasal mucosa proved to be more sensitive than oropharyngeal epithelia. The results demonstrate genotoxic effects of phthalates on human mucosal cells of the upper aerodigestive tract, in contrast to earlier findings in animal models. PMID- 10692223 TI - Use of catalytic topoisomerase II inhibitors to probe mechanisms of chemical induced clastogenicity in Chinese hamster V79 cells. AB - Determination of the clastogenic potential of new chemical entities, particularly pharmaceuticals, is an important part of the overall safety assessment of such drugs. It is appreciated that clastogenicity can arise from perturbation of many different cellular processes distinct from direct DNA/drug interactions. One such alternative clastogenic process is inhibition of DNA topoisomerase II, during which process the topoisomerase/DNA/drug ternary complex forms stable DNA double strand breaks (cleavable complex), which become templates for recombinational, mutagenic, and chromosomal fragmentation events. Without extensive experimentation, it is generally not possible to distinguish clastogenicity arising from direct drug/DNA interaction from that arising from inhibition of topoisomerase II. In the present investigation, we demonstrate that specific catalytic inhibitors of DNA topoisomerase II reduce the clastogenicity of topoisomerase poisons but not that arising via non-topoisomerase-dependent mechanisms. In particular, it is shown that catalytic topoisomerase II inhibitors such as chloroquine, sodium azide, and A-74932, as well as certain intercalating agents such as 9-aminoacridine and ethidium bromide, strongly antagonize the formation of micronuclei induced by the DNA gyrase inhibitor clinafloxacin and the antitumor topoisomerase II poison etoposide. These catalytic inhibitors are also shown to antagonize the clastogenicity of experimental compounds and novel pharmaceuticals presumed to be DNA intercalating agents by virtue of their response in a cell-based bleomycin amplification assay. We extend our previous hypothesis, suggesting that the clastogenicity of some nonstructurally alerting drugs may be due to an as yet unappreciated propensity for DNA intercalation. It is further proposed that intercalation-dependent inhibition of DNA topoisomerase II may be responsible for this clastogenicity and that this may be detected in intact mammalian cells with the use of catalytic topoisomerase inhibitors. PMID- 10692224 TI - Oxidative mutagenesis in Escherichia coli strains lacking ROS-scavenging enzymes and/or 8-oxoguanine defenses. AB - Escherichia coli strains with different combinations of null mutations in the katG, katE, sodA, sodB, fpg, and mutY genes were constructed to compare their spontaneous mutation frequencies and sensitivities to various oxidants with those of bacteria solely deficient in catalase (katG katE) or cytosolic superoxide dismutase (sodA sodB) and the parental strain possessing a full complement of these enzymes. The MutY DNA glycosylase represented the major protection against the mutagenic consequences of processes associated with normal aerobic metabolism. Spontaneous mutagenesis in MutY-lacking bacteria was not influenced by the absence of (A)BC excinuclease or the presence of MucAB proteins, a result consistent with 8-oxoguanine being a principal premutational lesion. In contrast, catalase and SOD represented the major protection against the genotoxic consequences of bursts of oxidative stress caused by reactive-oxygen-generating compounds. Therefore, only bacteria simultaneously defective in both katG and katE or sodA and sodB genes were hypersensitive with respect to mutability by peroxide and superoxide, respectively. These data suggest that oxidative lesions other than 8-oxoguanine contribute to mutagenesis by hydrogen peroxide and redox cycling chemicals. PMID- 10692225 TI - UV-induced mutagenesis of human p53: analysis using a double-selection method in yeast. AB - Comparison of the mutation patterns of p53 in human tumors with those of selectable genes in model systems is a powerful approach to identify potential etiological factors for specific tumor types. Recently, we validated use of a yeast assay to permit direct determination of the mutation spectrum induced in human p53 by carcinogens that would reduce uncertainties inherent in comparing spectra induced in different target genes. Here, we describe modifications in the assay designed to facilitate screening for mutants and to permit intracellular exposure of the gene instead of in vitro treatment. This was accomplished by introducing growth-based selection for transactivation-deficient p53 mutants into yeast already possessing red/white colony color selection. This improved model system was able to detect cells harboring p53 mutations among cells with wild type p53 at a frequency of 10(-4) or less. Additionally, UV light was used to verify that the majority of mutagenized cells with the appropriate phenotype on selective medium contained mutations in p53, not elsewhere in the genome. Sequence analysis of UV-induced mutations revealed that the nature of the mutations was similar to those obtained in previous studies of this mutagen. This system will prove useful in the determination of the ability of environmental agents to mutate the human p53 gene, and thus may contribute to hazard identification. PMID- 10692226 TI - Nitric oxide-induced mutations in the HPRT gene of human lymphoblastoid TK6 cells and in Salmonella typhimurium. AB - Characterization of mutations induced by NO in different experimental systems will facilitate elucidation of mechanisms underlying its genotoxicity. The mutagenic specificity of NO in human cells is of particular interest in view of its potential role in inflammation-associated carcinogenesis. We compared mutagenesis in human lymphoblastoid TK6 cells and in Salmonella typhimurium induced by exposure to NO delivered into the medium at rates approximating its production by activated macrophages. Exposure of TK6 cells continuously for 60 min decreased viability by 88%, and survivors exhibited a sixfold increase in mutant fraction in the hprt gene. Independent mutants were isolated and mutations characterized by RT-PCR and DNA sequencing. Among a total of 68 mutants analyzed, RT-PCR products were obtained in 41 (60%), and cDNA sequencing revealed that 26 (63%) of them contained mutations located in the hprt coding region. Base substitutions were present in 18 mutants, 12 occurring at A:T base pairs. Seven mutants contained deletions of 1-27 bp and one a 13-bp insertion; the 15 remaining RT-PCR products contained whole-exon deletions, 14 involving single exons. Six tester strains of S. typhimurium, each containing one of the six possible point mutations in the target codon of a gene in the histidine biosynthetic pathway, were similarly treated with NO and induction of mutation was detected by reversion to histidine auxotrophy. Significant increases were observed in frequencies of each of the six possible base mutations, with the highest occurring in G:C --> A:T transitions. The pattern of NO-induced hprt mutations in TK6 cells was similar to a recently published spectrum in spontaneous mutants, suggesting that reactive species derived from NO may contribute to spontaneous mutagenesis of the endogenous hprt gene in human cells. PMID- 10692227 TI - Mutagenic potential of adenine N(6) adducts of monoepoxide and diolepoxide derivatives of butadiene. AB - To determine the biological effects of specific DNA adducts resulting from the interaction of 1,3-butadiene metabolites with DNA, deoxyoligonucleotides have been synthesized with four different adducts at the N(6) position of adenine, centrally located within the human N-ras codon 61. The adducts are those arising from adduction by either the R or S stereoisomer of the monoepoxide (BDO) or the (R,R) or (S,S) isomer of the diolepoxide (BDE). The diolepoxide can arise from partial hydrolysis of the diepoxide (BDO(2)) or from epoxidation of hydrolyzed monoepoxide. These adducted oligonucleotides were used in in vivo and in vitro assays designed both to determine their mutagenic potency and to examine specific interactions with Escherichia coli polymerases. Each adducted oligonucleotide was ligated into a single-stranded vector M13mp7L2 that was subsequently used to transfect E. coli. The resulting mutagenic spectrum for these modified DNAs was stereoisomer specific. Both monoepoxide lesions were nonmutagenic, but the mutagenic spectra for the modified DNAs containing BDE adducts were stereoisomer specific. The mutations generated by adducts of the R,R enantiomer of the diolepoxide were exclusively A --> G, whereas adducts of the S,S enantiomer of the diolepoxide yielded exclusively A --> C mutations. None of the four modifications resulted in significant blocks to in vivo phage replication, as evidenced by no decrease in plaque-forming ability. Consistent with these data, when each of three purified E. coli polymerases was used to replicate DNAs containing these adducted deoxyoligonucleotides, the individual polymerases appeared to be virtually unaffected, such that all lesions were readily bypassed. Whereas previous animal model studies identified the mutagenic spectrum related to butadiene exposure, these studies begin to establish the specific lesions responsible for mutagenesis. This is the first report of stereoselectivity related to butadiene-induced mutagenesis. PMID- 10692228 TI - Genetically modified Chinese hamster ovary cells for investigating sulfotransferase-mediated cytotoxicity and mutation by 2-amino-1-methyl-6- phenylimidazo[4,5-b]pyridine. AB - To test the hypothesis that the sulfotransferase gene plays a role in the phase II bioactivation of PhIP, a heterocyclic amine found in cooked meats, we transfected the UV5P3 cell line with cDNA plasmids of human aryl sulfotransferases (HAST1 and HAST3). UV5P3 is a nucleotide excision repair deficient and P4501A2-expressing CHO cell line that we have previously developed. Functionally transformed clones were identified by the differential cytotoxicity (DC) assay that used PhIP as the cytotoxic agent. Two clones designated 5P3H1 and 5P3H3, expressing HAST1 and HAST3, respectively, were chosen for further characterization. Correct fragment sizes of the sulfotransferase cDNAs were identified in both cell lines by polymerase chain reaction. Immunoblot analysis confirmed the expression of the sulfotransferase proteins. The addition of the sulfotransferase inhibitor DCNP decreased the cytotoxic effects of PhIP in a dose dependent manner. The increase in cell growth was 6. 5-fold for 5P3H1 and 2.4 fold for 5P3H3, relative to values obtained without DCNP. Based on D(50) values, the dose that reduced the survival to 50% relative to untreated controls, the cytotoxic effect of PhIP was increased threefold for 5P3H1 and 1.87-fold for 5P3H3 cell lines over the parental UV5P3 line. There was also a small increase in the mutation response at the aprt locus. These newly established 5P3H1 and 5P3H3 sulfotransferase-expressing cells provide valuable mechanistic information of the bioactivation of PhIP and related compounds. Environ. Mol. Mutagen. 35:57-65, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10692229 TI - Effect of stable integration of the Escherichia coli ada gene on the sensitivity of Saccharomyces cerevisiae to the toxic and mutagenic effects of alkylating agents. PMID- 10692230 TI - New bioactive flavonoids and stilbenes in Cube resin insecticide PMID- 10692231 TI - Annocherin and (2,4)-cis- and trans-annocherinones, monotetrahydrofuran annonaceous acetogenins with a C-7 carbonyl group from annona cherimolia seeds PMID- 10692232 TI - Biased retellings of events yield biased memories. AB - When people retell events, they take different perspectives for different audiences and purposes. In four experiments, we examined the effects of this postevent reorganization of events on memory for the original events. In each experiment, participants read a story, wrote a biased letter about one of the story characters, and later remembered the original story. Participants' letters contained more story details and more elaborations relevant to the purpose of their retellings. More importantly, the letter perspective affected the amount of information recalled (Experiments 1, 3, and 4) and the direction of the errors in recall (Experiments 1 and 3) and recognition (Experiment 2). Selective rehearsal plays an important role in these bias effects: retelling involves selectively retrieving and using story information, with consequent differences in memory. However, biased memory occurred even when the biased letters contained little, if any, specific information (Experiment 4) or contained the same amount and kinds of story information as a neutral control condition (Experiment 3). Biased memory is a consequence of the reorganizing schema guiding the retelling perspective, in addition to the effects of rehearsing specific information in retelling. PMID- 10692233 TI - Representation versus process in simplicity of serial pattern completion. AB - This study deals with the question whether preferred extrapolations of serial patterns are determined by the extrapolation process or the representation of serial patterns. Three experiments are reported. The first deals with series of letters from the alphabet. Each series gave rise to extrapolations with varying frequencies, and we investigated how well these frequencies are predicted by the information loads, stemming from three models. The model of Klahr and Wallace (1970) is mainly concerned with the process of extrapolation. The model of Simon and Kotovsky (1963) describes both the process and the representation of extrapolated series. A third model is introduced here that exclusively deals with the representation of series. The outcome indirectly indicates that the preferred extrapolation is determined by the simplest representation of a series and not by the simplest process towards this representation. To strengthen the last conclusion two experiments are added dealing with bar graph stimuli. They are designed to experimentally disentangle representation and process complexity. In the second experiment the task was to judge the complexity of each series after having extrapolated the series. In the third experiment, subjects had to judge complexity of extrapolated series without the task to extrapolate the series. The predictions from the three models make plausible that the complexity judgments of second experiment reflect the process of extrapolation and the complexity judgments of the third experiment still reflect the representation of series. The latter results again support the assumption that the preferred extrapolation is determined by the simplest representation of a series. PMID- 10692234 TI - Low-dose IL-2 reduces lymphocyte apoptosis and increases naive CD4 cells in HIV-1 patients treated with HAART. AB - During HIV disease an increased in vitro apoptosis of peripheral blood mononuclear cells has been demonstrated. This can be reversed in vitro by interleukin (IL)-2. Recent trials with highly active antiretroviral therapy (HAART) and IL-2 in HIV-1-infected patients showed promising immunological and clinical results. Here we investigated the effects of subcutaneous low-dose IL-2 administration in combination with HAART on in vitro apoptosis and the relationship between apoptosis, CD4(+) counts, and HIV replication. Twenty-two asymptomatic HIV patients were randomized for HAART (arm I) and HAART plus IL-2 (arm II). Spontaneous apoptosis was decreased in both arms after 28 weeks of therapy but the reduction was highly significant only in arm II (P = 0.05 vs P = 0.001). As the percentage of apoptosis decreased, there was a significantly higher increase of both CD4(+) and CD4(+) naive T cells in arm II vs arm I. HIV plasma viremia was reduced in all patients after therapy. Our data suggest that intermittent therapy with low-dose subcutaneous IL-2 in addition to HAART induces a positive immunomodulation in asymptomatic HIV-infected patients. PMID- 10692235 TI - Functional subsets within clonally expanded CD8(+) memory T cells in elderly humans. AB - With advancing age, healthy humans frequently demonstrate large clonal expansions of CD8(+) T cells in the peripheral blood, which persist for long periods of time and appear to be maintained as a population of memory cells. We studied nine large T cell clones in five elderly individuals. We noted that in most cases the expanded clones were dominated by cells that did not express CD28, a pivotal molecule in T cell activation, and these clones proliferated poorly in culture. However, nearly all of the clonal expansions had CD28(+) fractions and some of these cells appeared to lose CD28 gene expression with stimulation in culture. CD28(+) cells demonstrated greater proliferation in both bulk and limiting dilution cultures compared to CD28(-) cells bearing the same TCR, whereas CD28(-) cells showed increased perforin expression. Together, these data suggest that loss of CD28 expression marks functional differentiation to cytotoxic memory cells within these clonal expansions and likely within CD8(+) memory populations in general. PMID- 10692236 TI - Human mast cells express the hyaluronic-acid-binding isoform of CD44 and adhere to hyaluronic acid. AB - CD44 is expressed in various isoforms on multiple cell lineages including those of hematopoietic origin and is believed in part to mediate cell adhesion to hyaluronic acid. Elevated levels of soluble CD44 (sCD44) have been identified in the serum of some patients with specific neoplasms. We thus sought to determine whether human mast cells express functional CD44 and whether sCD44 might be associated with systemic mast cell disease. Using a standard assay, CD34(+) derived cultured human mast cells were first demonstrated to adhere to hyaluronic acid-coated surfaces. Human mast cells were then found by flow cytometry to express CD44S, but not the v5, v6, v7, and v8 isoforms, and to shed CD44S following activation induced by PMA or aggregation of FcvarepsilonRI. However, CD44S was not found to be consistently elevated in serum obtained from patients with mastocytosis or individuals experiencing anaphylaxis. Thus, human cultured mast cells express and shed CD44S, which appears to mediate the attachment of these cells to hyaluronic acid. PMID- 10692238 TI - Fatal Mycobacterium bovis BCG infection in TNF-LT-alpha-deficient mice. AB - Neutralization of TNF or disruption of TNF-R1 leads to fatal Mycobacterium bovis BCG infection. Here we used TNF-LT-alpha-deficient mice to test whether a complete disruption of TNF and LT-alpha reduces further host resistance to BCG infection. The bacterial burden especially in the lungs of TNF-LT-alpha-deficient mice was significantly increased and the mice succumbed to infection between 8 and 10 weeks. In the absence of TNF-LT-alpha the granulomatous response was severely impaired and delayed. The cells in the granulomas of TNF-LT-alpha deficient mice expressed low levels of MHC class II and ICAM-1. They contained a few T cells and F4/80-positive macrophages expressing little iNOS and acid phosphatase activity. By contrast, the lethal action of endotoxin was dramatically reduced in BCG-infected TNF-LT-alpha-deficient mice. In summary, in the absence of TNF-LT-alpha the recruitment and activation of mononuclear cells in response to BCG infection were significantly delayed and reduced resulting in immature granulomas allowing uncontrolled fatal infection. PMID- 10692237 TI - NFAT1 enhances HIV-1 gene expression in primary human CD4 T cells. AB - Cyclosporin A (CsA) is a potent inhibitor of the NFAT family of transcription factors that enhance T cell activation. The observation that human immunodeficiency virus type 1 (HIV-1)-positive transplant recipients have a reduced HIV-1 viral burden during treatment with CsA suggested that NFAT may play a direct role in enhancing transcription of the HIV-1 viral genome. Two sets of NFAT binding sites were identified in the HIV-1 long terminal repeat (LTR) promoter by in vitro footprinting with full-length recombinant NFAT protein, and gel shift analysis of nuclear protein from polyclonally activated primary CD4 T cells revealed specific binding of NFAT1 to the NFkappaB binding sites of the HIV 1 LTR. Activation of primary CD4 T cells transiently transfected with a HIV-1 LTR luciferase reporter plasmid, lacking the NFAT binding sites in the upstream putative negative regulatory element but maintaining the NFkappaB/NFAT sites, demonstrated increased HIV-1 gene expression when cotransfected with a NFAT1 expression vector. Moreover, CsA, FK506, and a dominant-negative NFAT1 protein independently inhibited HIV-1 LTR promoter activity in CD4 T cells stimulated with phorbol ester and calcium ionophore. In primary human CD4 T cells, CsA also inhibited promoter activity directed by multimers of binding sites for NFAT, while having no effect on NFkappaB multimer-driven promoter activity. Increasing NFAT1 levels in CD4 T cells transiently transfected with a HIV-1 provirus also increased p24 protein expression. Thus, NFAT may be a target for prevention of HIV-1 LTR-directed gene expression in human CD4 T cells. PMID- 10692239 TI - IL-12-Dependent enhancement of CTL response to weak class I-restricted peptide immunogens requires coimmunization with T helper cell immunogens. AB - The effect of in vivo administration of rmIL-12 on the CTL response to immunization with a weakly immunogenic class I-restricted peptide emulsified in incomplete Freund's adjuvant was investigated. In the absence of IL-12, peptide specific CTL responses were significantly greater following coimmunization with class I-restricted peptide and T helper cell antigens than following immunization with the class I-restricted peptide alone. IL-12-dependent enhancement of the CTL response to peptide immunization was demonstrated in the presence of, but not in the absence of, coimmunization with T helper cell antigen. These findings indicate that IL-12 enhancement of the CTL response to weak class I-restricted immunogens is T helper cell dependent. Treatment with rmIL-12 also enhanced the CTL response to immunization with cDNA encoding both CTL and T helper cell epitopes. These findings are relevant to the design of vaccines containing tumor associated class I-restricted peptides currently being tested as an immunotherapy for cancer patients. PMID- 10692240 TI - Interleukin-6 release by cultured peripheral blood mononuclear cells inversely correlates with height velocity, bone age, insulin-like growth factor-I, and insulin-like growth factor binding protein-3 serum levels in children with perinatal HIV-1 infection. AB - Spontaneous and phytohemagglutinin (PHA)-stimulated interleukin (IL)-6 release by cultured peripheral blood mononuclear cells was related to height velocity, bone age, insulin-like growth factor-I (IGF-I), and IGF binding protein-3 (IGFBP-3) serum level standard deviation scores (SDS) of 32 children [aged 91 (median; range 13-151) months] with human immunodeficiency virus-type 1 (HIV-1) perinatal infection and severe disease. Spontaneous and PHA-stimulated IL-6 release inversely correlated with height velocity, bone age, IGF-I, and IGFBP-3 SDS. Ten children with height velocity SDS -2, showed higher spontaneous and PHA-stimulated IL-6 release and lower IGF-I and IGFBP-3 SDS (irrespective of CD4-positive T-lymphocyte counts, viral load, liver disease, or nutrition status). IL-6 overproduction may be a mechanism of IGF-I and IGFBP-3 down-regulation and impaired linear growth in children with perinatal HIV-1 infection. Growth-promoting strategies, including targeted anticytokine treatments, could be devised for such children. PMID- 10692245 TI - To be, or not to be, that is the question. Apoptosis in human trophoblast. AB - Apoptosis, the morphology of cell suicide, may result from programmed cell death or may be a response to exogenous stimuli. Apoptosis can be induced in cultured trophoblast and can be identified in the trophoblast of placental villi. The trophoblast regulates maternal-fetal gas, nutrient and waste product exchange; therefore, the presence of apoptosis in this key cellular interface highlights the importance of understanding what controls apoptosis in the placenta. In this review, we describe the signal transduction pathways that trigger apoptosis in other systems, identify key genetic controls for the process and outline the final common pathway which effects execution in cells committed to suicide. Multiplicity, redundancy and cross talk among pathways characterize the surface membrane signals and exogenous stimuli that trigger apoptosis in other cells. As each step in the apoptotic process is discussed, we describe what is known about the step in human placental villi. Recent studies suggest that a further understanding of receptor-mediated signalling pathways, the Bcl-2 regulators and the caspases and substrates involved in placental apoptosis will surely provide insights into both normal placental development and the placental dysfunction associated with some abnormal pregnancies. PMID- 10692246 TI - Glucose transporters in the human placenta. AB - The availability of antibodies and cDNA probes specific for the various members of the facilitated-diffusion glucose transporter (GLUT) family has enabled researchers to obtain a much clearer picture of the mechanisms for placental uptake and transplacental transport of glucose. This review examines studies of human placental glucose transport with the aim of providing a model which describes the transporter isoforms present in the placenta, their cellular localization and functional significance. The GLUT1 glucose transporter, present on both the microvillous and basal membranes of the syncytial barrier, is the primary isoform involved in the transplacental movement of glucose. Although GLUT3 mRNA is widely distributed, GLUT3 protein is localized to the arterial component of the vascular endothelium, where it may play a role in enhancing transplacental glucose transport. This data is in contrast to the situation in other mammalian species, such as the mouse, rat and sheep, where GLUT3 protein is not only present in those epithelial cells which carry out transplacental transport but becomes an increasingly prominent isoform as gestation progresses. The asymmetric distribution of GLUT1 in the human syncytiotrophoblast (microvillous>basal) means that basal GLUT1 acts as the rate limiting step in transplacental transfer. Changes in basal GLUT1 therefore have the potential to cause alterations in transplacental transport of glucose. Although there appear to be no changes in syncytial GLUT1 expression in intrauterine growth retardation, in diabetic pregnancies increases in basal GLUT1 expression and activity have been observed, with significant consequences for the maternal-fetal flux of glucose. Little is known of glucose transporter regulation in the placenta save for the effects of hyper- and hypoglycemia. GLUT1 expression and activity appear to be inversely related to extracellular glucose concentration, however within the physiological range, GLUT1 expression is relatively refractory to glucose concentration. Information is still needed on gestational development, on the expression and activity in well-defined conditions of intrauterine growth retardation, on the mechanisms and consequences of the changes observed in diabetic pregnancy and on the role of external agents other than glucose in regulating placental glucose transport. PMID- 10692247 TI - Expression of endothelial nitric oxide synthase by extravillous trophoblast cells in the human placenta. AB - Previous reports have documented the expression of endothelial nitric oxide synthase (eNOS) expression by the syncytiotrophoblast layer of the villus in the human placenta. In contrast, the underlying villous cytotrophoblast cells do not express the enzyme. Because extravillous cytotrophoblasts have not been as extensively investigated, our objective was to test whether these cells express eNOS. Using both a mouse monoclonal and a rabbit polyclonal antibody, we demonstrated immunoreactive eNOS in trophoblast cell columns emanating from anchoring villi in second trimester placentae. Cytokeratin positive trophoblast cells lying beneath remnant anchoring villi, lining decidual blood vessels and scattered throughout the basal plate of normal term and pre-eclamptic placentae also expressed immunoreactive eNOS. By Western analysis, the monoclonal and polyclonal antibodies were shown to be absolutely and relatively specific for eNOS, respectively. The finding of immunoreactive eNOS expression by extravillous trophoblast cells was substantiated by in situ hybridization. Using riboprobes generated from a bovine eNOS cDNA, we demonstrated specific hybridization in the endothelium of blood vessels in the umbilical cord, thus validating the in situ hybridization methodology, as well as specific hybridization in the extravillous trophoblast cells of the basal plate in normal term placenta. In conclusion, several different populations of extravillous trophoblast cells in the basal plate of the human placenta express eNOS. PMID- 10692248 TI - Human placenta, chorion, amnion and decidua express different variants of corticotropin-releasing factor receptor messenger RNA. AB - Human placenta is a major source of corticotropin-releasing factor (CRF), and local effects of CRF in fetal membranes and placenta have been shown, i.e., adrenocorticotropic hormone (ACTH) and oxytocin release from cultured placental cells, as well as prostaglandin release from amnion, chorion and decidua. Two distinct CRF receptors (CRF-R1 and CRF-R2) have been characterized: CRF-R1 consists of two isoforms (CRF-R1alpha and CRF-R1beta) while CRF-R2 has at least three different splice variants (CRF-R2alpha, CRF-R2beta and CRF-R2gamma). To date, CRF-R1 receptor has been identified in human placenta and in pregnant myometrium, while no evidence for placental CRF-R2 receptor isoforms has been provided. The present study investigated whether the different isoforms of CRF-R1 and CRF-R2 receptor mRNA are expressed in fetal membranes and placenta. Tissues were collected after spontaneous vaginal delivery (38-40 weeks) or elective caesarean section (39-41 weeks). The gene expression of CRF receptors was first studied by reverse transcriptase-polymerase chain reaction (RT-PCR), and the presence of CRF-R1alpha, but not of CRF-R1beta, in human placental trophoblast, amnion/chorion and decidua was shown. In addition, among the three CRF-R2 splice variants, only CRF-R2beta mRNA was expressed by trophoblast and fetal membranes. By using in situ hybridization, CRF-R1 and CRF-R2 probes positively hybridized trophoblast and related membranes. CRF-R1 was localized in the syncytiotrophoblast cells, chorionic trophoblast and decidua with a small amount in the amnion. CRF-R2 probe mainly hybridized syncytiotrophoblast cells, but cytotrophoblast also contained discreet amounts of CRF-R2 mRNA signal. The CRF-R2 hybridization signal was also observed within the structure of the villi (blood vessels), chorionic trophoblast and decidual cells, but it was faint or absent in the amniotic epithelium. There was no significant difference in the distribution of CRF-R1 or CRF-R2 mRNA signal between placentas collected from vaginal delivery or caesarean section. The evidence that intrauterine tissues differently express CRF-R1alpha and CRF-R2beta supports possible different local roles of CRF and related peptides into intrauterine tissues during pregnancy. PMID- 10692249 TI - Activin A exerts both pro- and anti-inflammatory effects on human term gestational tissues. AB - There is a growing appreciation of the importance of activin as a modulator of immune function. The aim of the present study was to determine whether activin A exerts any effects on cytokine and prostaglandin (PG) production by the tissues of pregnancy. Explant cultures were established for amnion, choriodecidual and placental tissues derived from pregnancies delivered at term by Caesarean section (n=5 placentae). Explants were treated with activin A (0.5, 5 and 50 ng/ml) in serum-free Ham's F12/DME media for 24 h (n=3-4 replicates). Production rates of interleukin (IL)-1beta, IL-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and PGE(2)were determined using immunoassay. Differences between treatment groups were analysed by ANOVA followed by Dunnett's test;P< 0.05 was considered to be significant. Amnion IL-6 production exhibited biphasic responses to activin A: at 5 ng/ml activin A, IL-6 production was significantly stimulated (to 246+/-74.6 per cent of control (mean+/-sem), while at 50 ng/ml it was significantly inhibited (to 46+/-7.4 per cent of control). IL-8 and PGE(2)production by amnion showed significant responses to activin A that were similar to those of IL-6. No significant effects of activin A were observed on choriodecidual and placental IL 6, IL-8 and PGE(2)production. However, TNF-alpha production was significantly inhibited by 50 ng/ml activin A in both choriodecidual and placental explants (to 43+/-9.7 per cent and 51+/-6.7 per cent of control, respectively). Placental IL 1beta production was not altered by treatment with activin A at any concentration. These findings support the concept of activin as an immune modulator in tissues of pregnancy. PMID- 10692250 TI - Myofibroblast differentiation in the connective tissues of the amnion and chorion of term human fetal membranes-implications for fetal membrane rupture and labour. AB - An area of the fetal membranes, within the rupture tear after spontaneous delivery at term, exhibits altered morphology compared to more distal sites. It is characterized by marked swelling of the amniotic and chorionic connective tissue layers, consistent with structural weakness, and a marked reduction of the thickness of both the cytotrophoblast and decidual layers. These features, albeit less extreme, have been identified in fetal membranes in the lower uterine pole in patients prior to labour. In this study of pre-labour, labour-affected and post-labour term fetal membranes, we report that these regions are associated with an alteration in the phenotype of the vimentin positive mesenchymal cell population of the chorionic connective tissue reticular layer, and are consistent with myofibroblastic differentiation, i.e. alpha-smooth muscle actin (alpha-sma) expression. In the reticular layer of the lower uterine pole biopsies in the labour-affected group the numbers and densities of alpha-sma immunoreactive positive cells were 17-fold (P=0.04) and 8.5-fold (P=0.02) higher than in mid zone biopsies. After delivery, in rupture line biopsies the numbers and densities were 50-fold (P=0. 002) and 36-fold (P=0.003) higher compared to mid zone biopsies. The percentage of the vimentin positive population positive for alpha sma was 2-5 per cent in mid-zone biopsies compared to 49 per cent (P=0.03) in the labour-affected 'cervical' biopsies and 69 per cent (P=0.05) in the rupture line biopsies. Within the tear sites, alpha-sma positive cells were also detected within the fibroblastic layer of the amniotic connective tissue. Although there was no significant difference between the numbers and density of alpha-sma cells in the reticular layers between mid and lower uterine pole biopsies in the pre labour group, in a proportion of patients the biopsies were similar to labour affected biopsies indicating that this alteration occurs prior to clinically apparent labour in these patients. The incidence of alpha-sma positive cells in the reticular layer correlated with morphological changes within the fetal membranes, for example thickness of reticular (r(2)=0.349, P=0.0006) and amniotic connective tissue layers (r(2)=0.389, P=0.0002). This suggests that cellular activities associated with myofibroblastic differentiation in the reticular layer of the chorion may be associated with the observed connective tissue changes, fetal membrane rupture and labour. PMID- 10692251 TI - The roles of the cyclo-oxygenases types one and two in prostaglandin synthesis in human fetal membranes at term. AB - The aim of this study was to determine the relative contributions of cyclo oxygenase (COX) types 1 and 2 to prostaglandin synthesis at term. METHODS: Fetal membranes were collected from 6 pregnancies after elective caesarean section at term, prior to labour. The presence of COX-1 and COX-2 protein was determined using Western analysis. The relative contributions of the two isoforms of COX to prostaglandin synthesis were determined by incubation of fetal membrane discs with either a COX-2 selective inhibitor, SC236, or a COX-1 selective inhibitor, SC560, and measurement of prostaglandin release during 24 h using enzyme-linked immuno-sorbent assay (ELISA). RESULTS: Both COX-1 and COX-2 protein were demonstrated in amnion and chorion-decidua. The COX-2 selective inhibitor, SC 236, significantly reduced prostaglandin synthesis, both in its COX-2 specific and higher, non-specific concentration ranges. The COX-1 selective inhibitor, SC 560, had no effect upon prostaglandin synthesis in its COX-1 specific concentration range, but did significantly reduce prostaglandin synthesis at higher, non-selective concentrations. CONCLUSIONS: Fetal membranes contain both COX-1 and COX-2 at term, but only COX-2 contributes towards prostaglandin synthesis. COX-2 selective NSAI drugs will be as effective as non-selective agents in inhibition of fetal membrane prostaglandin synthesis and may represent a new strategy for tocolysis. PMID- 10692252 TI - Expression of the p53 gene and apoptosis in gestational trophoblastic disease. AB - In order to understand the involvement of the p53 tumour suppressor gene in the pathogenesis of gestational trophoblastic disease (GTD), we investigated its genetic status, protein expression and its role in apoptosis in samples of complete and partial hydatidiform mole as compared with those of normal placenta. Direct sequencing of polymerase chain reaction (PCR) products of the coding and non-coding regions of the p53 gene demonstrated no mutations in any of the studied samples. Immunohistochemical studies revealed increased expression of the p53 protein predominantly in the nuclei of villous cytotrophoblasts. This over expression of p53 was found in all samples of complete mole, in 50 per cent of partial mole samples and in about 30 per cent of normal placenta cases, although no significant difference in the staining intensity and pattern was observed. An in situ detection of DNA nicking (TUNEL) staining, demonstrating apoptosis, was also detected predominantly in villous cytotrophoblasts and in stromal areas. The per centage of apoptotic cells in all studied samples, determined by flow cytometry, demonstrated a significant increase in apoptotic cells in samples of complete and partial hydatidiform mole compared with those of normal placenta (P< 0.0003 and P< 0.004, respectively). In conclusion, the current study may provide a possible explanation to the pathogenesis of GTD, probably associated with extensive p53-dependent apoptosis to modulate excessive trophoblastic proliferation. PMID- 10692253 TI - Expression of aminopeptidase A in human gestational choriocarcinoma cell lines and tissues. AB - Aminopeptidase A (AP-A), a cell-surface metallopeptidase hydrolyzing peptide with N-terminal acidic residues, has been proved to be identical to the B cell differentiation antigen BP-1 and to the kidney differentiation antigen gp160, suggesting recognition of AP-A as a differentiation-related marker on certain normal and transformed cells. AP-A has also been purified from human placenta and been shown to be localized in the trophoblasts. In the present study, we examined the expression and enzymatic activity of AP-A in human gestational choriocarcinoma, a neoplastic transformant from trophoblasts which comprises a heterogenous population of trophoblastic cells in different stages of differentiation. Flow cytometry and immunoblot analysis demonstrated that AP-A was expressed in five choriocarcinoma cell lines which were secreting low or moderate levels of human chorionic gonadotropin (hCG), while two high hCG secreting cell lines lacked AP-A expression. The AP-A enzymatic activity correlated with cell-surface levels of AP-A and was abrogated by amastatin, an inhibitor of AP-A. Immunohistochemical analysis revealed that AP-A was present in seven of eight choriocarcinoma tissues and was localized on the cell membrane of cytotrophoblastic choriocarcinoma cells, but not on cells with syncytiotrophoblast-like features. These results demonstrate that AP-A is expressed on most choriocarcinomas and its expression is restricted to low hCG secreting, cytotrophoblastic cells and down-regulated as a function of cell differentiation, suggesting an involvement of AP-A in the differentiation/maturation process of neoplastic trophoblasts. PMID- 10692254 TI - The cellular mechanism by which the human endogenous retrovirus ERV-3 env gene affects proliferation and differentiation in a human placental trophoblast model, BeWo. AB - The env region of the human endogenous retrovirus ERV-3 is expressed during differentiation of trophoblast and the choriocarcinoma BeWo. Stable transfectants with ERV-3 env exhibit most aspects of trophoblast differentiation, including inhibition of cell proliferation, changes in cell morphology, and increased production of beta-hCG mRNA. In this study, the cellular mechanism of induction of BeWo cell differentiation by ERV-3 env was investigated. In BeWo cells stably transfected with ERV-3 env, the production of beta-hCG mRNA and hCG protein was increased. Intracellular cAMP level was markedly increased over that of vector transfected cells. The effect on beta-hCG protein production was inhibited by H89, a protein kinase A (PKA) inhibitor, while protein kinase C (PKC) and protein tyrosine kinase (PTK) inhibitors had no effect. The expression of a major cell cycle promoter, cyclin B, was markedly reduced while expression of p21, a negative regulator of the cell cycle, was up-regulated. Inhibition of ERV-3 env induced hCG production with H89 had no significant effect on cell growth when compared with cells transfected with vector alone. PMID- 10692255 TI - The molar vesicle fluid contains the beta-core fragment of human chorionic gonadotropin. AB - The human chorionic gonadotropin (hCG) beta-core fragment (beta-CF) is a major molecular form of hCG beta subunit (hCGbeta) immunoreactivity in the urine of pregnant women and patients with trophoblastic disease. The majority of evidence supports the fact that the beta-CF is a degradative product of intact hCG and free hCGbeta in the kidneys. We found a beta-CF-like substance in the fluid of molar vesicles from a patient with complete hydatidiform mole. The molar fluid beta-CF (mbeta-CF) was indistinguishable from the beta-CF in the patient's urine (ubeta-CF) by immunoreactivity and by elution profile on gel chromatography. The binding study to lectins, however, showed that mbeta-CF contains a carbohydrate moiety that differs from that of ubeta-CF. Immunohistochemistry with anti-beta-CF antibody demonstrated a strong immunoreactivity in a large number of macrophages in the molar villous stroma. In vitro incubation of intact hCG with peritoneal macrophages showed a slow increase of intact hCG in the cell cytosol with the appearance of beta-CF-like substance in the cell supernatant. In conclusion, the source of beta-CF in molar fluid is likely to be macrophages existing in the villous stroma. Thus macrophages may ingest intact hCG and act as a local regulator of gonadotropic hormones. PMID- 10692256 TI - Ontogeny of aquaporins 1 and 3 in ovine placenta and fetal membranes. AB - A sensitive and highly reproducible method has been used to show that Aquaporin 3 (AQP(3)) mRNA is present in the ovine placenta and chorion from at least 60 days of gestation (term=145-150d) with levels increasing substantially (>16 fold) at 100 days, and remaining constant thereafter. By immuno- and hybridization histochemistry, the epithelial cells expressing AQP(3)were found to be the trophoblast cells. Some AQP(3)was expressed in fibroblasts of the amnion and allantois but none was expressed in the epithelia of these membranes. AQP(1)was expressed in endothelial cells of fetal and maternal blood vessels but not in any epithelial cell of the ovine placenta and fetal membranes. The level of AQP(3)expression is consistent with known ovine placental permeabilities to water, glycerol and urea. PMID- 10692257 TI - Relationship between nutritionally-mediated placental growth restriction and fetal growth, body composition and endocrine status during late gestation in adolescent sheep. AB - The aim was to investigate the consequences of nutritionally-mediated placental growth restriction on fetal organ growth, conformation, body composition and endocrine status during late gestation. Embryos recovered from superovulated adult ewes inseminated by a single sire were transferred in singleton to the uterus of peripubertal adolescent recipients. Post-transfer, adolescent dams were offered a high (H) or moderate (M) level of a complete diet to promote rapid or moderate maternal growth rates, respectively (n=7 per group). After day 100 of gestation the feed intake of the M dams was adjusted weekly to maintain body condition score. Liveweight gain during the first 100 days of gestation was 301+/ 24 and 90+/-4.6 g/day for the H and M groups, respectively. Maternal plasma concentrations of insulin, IGF-I and urea were significantly higher and non esterified fatty acid concentrations significantly lower in H compared with M dams prior to slaughter on day 128 of gestation. At this stage of gestation, total placentome weight was 50 per cent lower in H compared with M groups (P< 0.001) and was associated with a 37 per cent reduction in fetal weight (P< 0.01). All variables of fetal conformation and absolute fetal organ weights, with the exception of the adrenal glands, were lower (P< 0. 05) in the fetuses from H intake dams. However, relative fetal organ weights expressed as g/kg fetal body weight, with the exception of the gut, were not influenced by maternal dietary intake. Furthermore, fetal weight but not maternal nutritional group were predictive of individual organ weight for all organs dissected. Together these results imply that growth restriction in the fetuses derived from H intake dams was largely symmetrical. Fetal plasma concentrations of insulin, IGF-I and glucose were attenuated (P< 0.05) in fetuses from H compared with M groups. The lower fetal body weight in the former group was associated with a reduction in absolute but not relative crude protein (P< 0.01) and fat content (P< 0.05). Total fetal liver glycogen content but not concentration was (P< 0.05) reduced in H versus M groups. The lower mass of both the placenta and fetal liver was due to a reduction in cell number rather than an alteration in cell size. Thus, over nourishing adolescent sheep is associated with a major restriction in placental growth which mediates a gradual slowing of fetal growth during the final third of pregnancy. PMID- 10692258 TI - Transfer and metabolism of prostaglandin E(2)in the dual perfused human placenta. AB - Prostaglandins (PGs) are potent paracrine hormones that are important for the control of several functions in the uterus and fetus during pregnancy and parturition. PGs are rapidly metabolized to inactive metabolites by prostaglandin dehydrogenase (PGDH). However, the regulation of transfer and metabolism of PGs across the placenta is not well understood. This study used an in vitro dual perfused human placental cotyledon preparation to examine the production of the potent vasoactive and myometrial stimulants PGE(2)and PGF(2alpha), transfer of PGs from the maternal to the fetal circulation and the metabolism of PGs by PGDH. Secretion of PGE(2)was greater into the fetal compared to the maternal circulation. PGE(2)output was higher than PGF(2alpha)and concentrations of PGE(2)and PGF(2alpha)metabolites (PGEM and PGFM) were greater in both fetal and maternal outputs when compared to the primary prostaglandins. Infusion of PGE(2)into the maternal circulation did not result in increased PGE(2)efflux but PGEM was output was increased, demonstrating a rapid and efficient metabolism by the placenta. There was no significant transfer of PGE(2)across to the fetal circulation, although there was some transfer but in the form of inactivated PGEM. There was no significant interconversion of PGE(2)to PGF(2alpha)by the 9 keto-reductase pathway. Expression of PGDH as detected by immunoblot was high in placenta. This PGDH was localized throughout the syncytiotrophoblast at the fetal maternal interface and also in extravillous trophoblast cells. The presence of PGDH at this site acts to stabilize output of primary PG from the placenta and also as a barrier preventing transfer to the fetal circulation, resulting in the separation of PG homeostasis in the fetus and mother. PMID- 10692259 TI - Enhanced expression of intercellular adhesion molecule-1 (ICAM-1) in amnion with term and preterm labour. AB - To evaluate the association between intercellular adhesion molecule-1 (ICAM-1) in the amnion and preterm labour and delivery, we have assessed ICAM-1 mRNA abundance by Northern analysis and protein levels by enzyme-linked immunosorbent assay (ELISA), in samples of this tissue after term and preterm delivery. The median ICAM-1 mRNA expression following preterm delivery (PTD, n=30) was 24 times greater (P< 0.05) than following elective caesarean section prior to labour at term (CST, n=14). ICAM-1 expression following vaginal delivery after spontaneous labour at term (SLT, n=11) was seven times greater than in the CST group (P< 0.05). The concentration of ICAM-1 protein in the PTD samples (n=31) was four fold greater than (P< 0.05) in CST (n=14). It was also three-fold greater than in the SLT (n=15) samples (P< 0.05). The results were substantially the same when a preterm spontaneous labour group (PTL) (n=26), exclusive of deliveries complicated by pre-eclampsia (n=1) or intrauterine growth restriction (n=3), was compared to the CST and SLT groups. The ICAM-1 mRNA expression did not differ significantly (P=0.93) between PTL with (n=12) or without (n=14) indicators of intrauterine infection. The results were similar when ICAM-1 protein concentrations were compared (P=0.43) between these two groups. These findings indicate that ICAM-1 is expressed by the human amnion and that this expression is elevated with preterm labour and delivery. PMID- 10692260 TI - The human placenta is encircled by a ring of smooth muscle cells. AB - The marginal zone of the human term placenta was studied by transmission electron microscopy and immunohistochemistry using antibodies against cytoskeletal filaments, extracellular matrix molecules and endothelial markers. The marginal sinus of the intervillous space is separated from the chorionic and basal plates by a layer of cells expressing vimentin, desmin, alpha- and gamma-smooth muscle actins, and smooth muscle myosin. Also ultrastructurally, these cells share all features with smooth muscle cells. This muscular ring is continuous with the media of uteroplacental veins entering the marginal sinus. In the basal plate the muscle cells may extend far into the central parts of the placenta. The muscular ring is separated from the intervillous space by a layer of endothelial cells. They are continuous with the maternal endothelium of the marginal uteroplacental veins. Moreover this endothelium covers neighbouring parts of the chorionic and basal plates, locally extending to the surfaces of large stem villi. The data suggest (1) that the marginal zone of the intervillous space ('marginal sinus') represents the dilated and merged parts of uteroplacental veins and (2) that lateral growth of the human placenta partly takes place by expansion into the uteroplacental veins. The functional importance of this muscular ring remains unknown. PMID- 10692261 TI - The effect of oxytocin, prostaglandin E2 and acetylsalicylic acid on flow distribution and on the transfer of alanine, glucose and water in isolated perfused guinea pig placentae. AB - The influence of oxytocin (OXY), sulproston (SUL) and acetylsalicylic acid (ASA) on L-alanine- (ALA), D-glucose- (GLU) or water- (H(2)O) uptake (maternal side) in the isolated perfused guinea pig placenta was investigated. Uptake was measured with a single injection, paired tracer dilution method. 'T50' values were derived from venous concentration curves (extracellular marker) as the distance (sec) between two concentration values at 50 per cent of peak concentration. T50 values were regarded to reflect the change of flow distribution on the maternal side. On average, there was a significant apparent inhibition of GLU uptake (by 27.2 per cent from control values) by OXY as well as of ALA uptake by OXY (26. 0 per cent), by ASA (56.6 per cent), and by SUL (56.7 per cent). The respective mean T50 values decreased significantly in the above groups by 15.9 per cent, 18.7 per cent (ns), 42.2 per cent and 56.7 per cent. However, it was not possible to generate dose-response curves whereas significant correlations of uptake values with T50 values were found. There was no dose-response relationship between T50 values and OXY or ASA concentrations but decreased mean T50 values were found. For SUL a weak correlation of T50 and SUL concentration was found. The r -value of GLU uptake and T50 was 0.57, for H(2)O uptake this value was 0.70, for ALA uptake the r -values were 0.51 (OXY), 0.35 (SUL) and 0.31 (ASA). Correlation of uptake and concentrations were not significant. We conclude that the 'inhibitory' effects of OXY, ASA and probably SUL on placental transfer are unspecific and the consequence of flow shifts from the placental exchange area to the uterine muscle. PMID- 10692262 TI - Advanced glycation end products: a Nephrologist's perspective. AB - Advanced glycation end products (AGEs) are a heterogeneous group of molecules that accumulate in plasma and tissues with advancing age, diabetes, and renal failure. There is emerging evidence that AGEs are potential uremic toxins and may have a role in the pathogenesis of vascular and renal complications associated with diabetes and aging. AGEs are formed when a carbonyl of a reducing sugar condenses with a reactive amino group in target protein. These toxic molecules interact with specific receptors and elicit pleiotropic responses. AGEs accelerate atherosclerosis through cross-linking of proteins, modification of matrix components, platelet aggregation, defective vascular relaxation, and abnormal lipoprotein metabolism. In vivo and in vitro studies indicate that AGEs have a vital role in the pathogenesis of diabetic nephropathy and the progression of renal failure. The complications of normal aging, such as loss of renal function, Alzheimer's disease, skin changes, and cataracts, may also be mediated by progressive glycation of long-lived proteins. AGEs accumulate in renal failure as a result of decreased excretion and increased generation resulting from oxidative and carbonyl stress of uremia. AGE-modified beta(2)-microglobulin is the principal pathogenic component of dialysis-related amyloidosis in patients undergoing dialysis. Available dialytic modalities are not capable of normalizing AGE levels in patients with end-stage renal disease. A number of reports indicated that restoration of euglycemia with islet-cell transplantation normalized and prevented further glycosylation of proteins. Aminoguanidine (AGN), a nucleophilic compound, not only decreases the formation of AGEs but also inhibits their action. A number of studies have shown that treatment with AGN improves neuropathy and delays the onset of retinopathy and nephropathy. N Phenacylthiazolium bromide is a prototype AGE cross-link breaker that reacts with and can cleave covalent AGE-derived protein cross-links. Thus, there is an exciting possibility that the complications of diabetes, uremia, and aging may be prevented with these novel agents. PMID- 10692263 TI - ACE inhibition improves glomerular size selectivity in patients with idiopathic membranous nephropathy and persistent nephrotic syndrome. AB - Patients with idiopathic membranous nephropathy (IMN) and persistent nephrotic range proteinuria are at risk for progression to end-stage renal failure. Whether angiotensin-converting enzyme (ACE) inhibitors are also renoprotective in these patients remains elusive. In 14 patients with IMN (patients) and persistent proteinuria (protein > 3 g/24 h for >6 months), we studied mean arterial pressure (MAP), urinary protein excretion, glomerular filtration rate (GFR), renal plasma flow (RPF), and albumin and neutral dextran fractional clearance after 2 months washout from previous antihypertensive treatment (basal), after 2 months of enalapril (2.5 to 20 mg/d) therapy (posttreatment), and 2 months after enalapril withdrawal (recovery). MAP, proteinuria, and GFR were also measured at the same time points in 6 patients with IMN and persistent overt proteinuria maintained on conventional treatment throughout the study period (controls). Basal MAP, proteinuria, and GFR were similar in the two study groups. However, in patients at the end of the treatment period, MAP (posttreatment, 99.6 +/- 11.2 versus basal, 103.3 +/- 12.1 mm Hg; P < 0.05), proteinuria (posttreatment protein, 5.0 +/- 2.9 versus basal, 7.1 +/- 4.9 g/24 h; P < 0.05), albumin fractional clearance (posttreatment median, 1.7 x 10(-3); range, 0.2 to 22.7 x 10(-3) versus basal median, 4.1 x 10(-3); range, 0.4 to 22. 1 x 10(-3); P < 0.05), and fractional clearance of largest neutral dextrans (radii from 62 to 66 A) were significantly less than basal values. At recovery, MAP significantly increased to 106.6 +/- 11.7 mm Hg (P < 0.001 versus enalapril), but all other parameters remained less than basal values. GFR and RPF were similar at each evaluation. Changes in proteinuria after treatment withdrawal positively correlated (r = 0.72; P < 0.01) with baseline GFR. Theoretical analysis of dextran-sieving data indicated that ACE inhibitor treatment significantly improved glomerular membrane size-selective dysfunction. This effect persisted more than 2 months after treatment withdrawal. No patient had symptomatic hypotension, acute renal function deterioration, or hyperkalemia during enalapril treatment. Thus, in patients with IMN and long-term nephrotic syndrome, ACE inhibitor treatment, but not conventional therapy, improves glomerular barrier size selectivity. The antiproteinuric effect of ACE inhibition is long lasting, especially in patients with more severe renal insufficiency. This is the premise of a long-term renoprotective effect that may limit the need for treatment with more toxic drugs. PMID- 10692264 TI - Outcome of IgA nephropathy in adults graded by chronic histological lesions. AB - This prognostic study of primary immunoglobulin A (IgA) nephropathy focused on chronic irreversible glomerular sclerosis and interstitial fibrosis, based on the premise that this disease is characterized by a protracted and, for many, progressive course. We used a chronicity-based histological grading system to assess the biopsy specimens of 126 adults with IgA nephropathy over a median follow-up of 10 years. Our grading system included a glomerular grading (GG) of 1 to 3 based on the extent of glomerular sclerosis, a tubulointerstitial grading (TIG) of 1 to 3 based on the degree of tubular loss or interstitial fibrosis, and the evaluation of hyaline arteriolosclerosis (HA). These three histological parameters were correlated with each other and with serum creatinine level, degree of proteinuria, and blood pressure at the time of renal biopsy. Univariate analysis showed that these three histological and three clinical parameters were significantly correlated with renal survival. By multivariate analysis using the Cox regression model, GG, serum creatinine level, and degree of proteinuria represented independent prognostic factors of renal survival. For a subset of patients at a relatively early stage of disease with a serum creatinine level less than 130 micromol/L at the time of biopsy, all three histological features and degree of proteinuria were significantly correlated with renal survival, and GG was the only independent prognostic factor for renal outcome. This study shows that glomerular sclerosis represents the most important prognostic factor in adult patients with primary IgA nephropathy and has a strong predictive value. Our chronicity-based histological grading system not only correlates well with the natural history of IgA nephropathy but is also reproducible and relatively simple to apply. PMID- 10692265 TI - Complement activation through the lectin pathway in patients with Henoch Schonlein purpura nephritis. AB - Henoch-Schonlein purpura nephritis (HSPN) is considered a form of systemic vasculitis of the small blood vessels with immune pathogenesis. In this disorder, the complement system is recognized as an important mechanism of glomerular injury. The aim of this study is to determine whether the lectin pathway, a novel pathway of complement activation, is related to the pathogenesis of HSPN. Renal biopsy material from 10 patients with HSPN was studied immunohistochemically and examined for a clinicopathologic correlation. Serum levels of complement components, including mannose-binding lectin (MBL), and plasma levels of complement activation products were also evaluated in these patients and compared with levels in patients with immunoglobulin A (IgA) nephropathy or mesangial proliferative glomerulonephritis (GN) without IgA deposition (non-IgA GN). Glomerular deposition of components of the pathway, MBL and MBL-associated serine protease (MASP-1), as well as C3b/C3c, C5b-9, and C4-binding protein (C4-bp), was detected in 8 of 10 patients. Although no significant correlation was found between glomerular deposition of MBL/MASP-1 and histological or clinical findings, the biopsies on all patients with MBL/MASP-1 deposits were performed within 20 weeks from the onset of disease. Levels of plasma C4d, the activation fragment of C4, and C4-bp, a soluble regulatory protein of the pathway, were greater in patients with HSPN than in those with non-IgA GN. However, there was no difference in serum MBL levels between the three groups of patients (HSPN, IgA nephropathy, and non-IgA GN). These results suggest that complement activation through the lectin pathway was involved at the onset of HSPN, and this mechanism might be important in the disease pathogenesis. PMID- 10692266 TI - Role of T lymphocytes in renal disease in HIV-transgenic mice. AB - The pathogenesis of human immunodeficiency virus (HIV)-associated focal segmental glomerulosclerosis (FSGS) has remained obscure. It has been proposed that renal parenchymal cells may be infected with HIV-1. If such infection occurs, the target cells would be expected to express viral proteins and thus could be targets for cytotoxic T lymphocytes. We previously described mice transgenic for a gag-pol-deleted HIV-1 genome that developed FSGS. In the present study, we tested the requirement for functional T cells in the evolution of renal disease in this model. We bred the HIV-transgenic mice (T26) with athymic nude mice to produce athymic T26 mice. We confirmed by flow cytometry of peripheral blood, thymus, lymph node, and spleen that the athymic T26 mice lacked mature T cells. The athymic T26 mice developed renal disease characterized by FSGS, tubular atrophy and dilatation, and interstitial infiltrate that was qualitatively identical to that seen in the parental T26 mice. Quantitative assessment of the athymic T26 mouse kidneys showed that glomerulosclerosis, tubular injury, and interstitial infiltrate were less severe compared with the parental T26 mouse kidneys. Although T26 mouse kidneys had a mixed cellular infiltrate composed of CD4 cells, CD8 cells, and macrophages, interstitial infiltrates within the athymic T26 mouse kidneys included macrophages but lacked both CD4 and CD8 cells. The renal expression of the HIV transgene was 1. 7-fold greater in T26 mice compared with athymic T26 mice. We conclude that mature T cells are not absolutely required for the development of HIV-associated nephropathy in transgenic mice but that, in their absence, renal disease is significantly milder. These data suggest that T-cell-mediated cytotoxicity directed against renal cells expressing virally encoded proteins is not an essential feature of renal pathogenesis in this model. PMID- 10692267 TI - Characteristics of albumin processing during renal passage in anti-Thy1 and anti glomerular basement membrane glomerulonephritis. AB - Recent studies have shown that glomerular-filtered albumin appears to be processed by two distinct cellular pathways. The major pathway, a high-capacity retrieval pathway, returns most of the filtered albumin to the blood supply intact. The albumin not taken up by the retrieval pathway is degraded by lysosomes during renal passage and excreted as fragments in urine. We studied the interplay of the albumin retrieval pathway and the degradation pathway in the disease models of anti-Thy1 nephritis, a model of mild proteinuria, and anti glomerular basement membrane (anti-GBM) disease, a model of severe proteinuria. This is achieved by investigating the integrity of urinary albumin and its excretion rate. Total albumin excretion (intact plus fragments) did not change significantly in the rats with anti-Thy1 nephritis. However, it was established that intact albumin excretion had a strong positive correlation with increasing total-protein excretion, which showed that the degradation pathway was being predominantly affected in this disease. For the rats with anti-GBM disease, total protein excretion increased 26-fold compared with the control group, and intact albumin excretion increased 250-fold. The profound changes in albumin excretion in anti-GBM disease are consistent with inhibition primarily of the retrieval pathway. PMID- 10692268 TI - Effect of antihypertensive therapy on renal function and urinary albumin excretion in hypertensive patients with autosomal dominant polycystic kidney disease. AB - Hypertensive patients with autosomal dominant polycystic kidney disease (ADPKD) have a faster progression to end-stage renal disease (ESRD) than their normotensive counterparts. The aim of this prospective, randomized study is to compare the effects of the calcium channel blocker amlodipine and the angiotensin converting enzyme inhibitor enalapril as first-line therapy on blood pressure, renal function, and urinary albumin excretion in hypertensive patients with ADPKD. Twenty-four patients with ADPKD with hypertension with creatinine clearances (Ccrs) greater than 50 mL/min/1.73 m(2) were included in the study. Twelve patients received amlodipine (mean dose, 9 mg/d), and 12 patients received enalapril (mean dose, 17 mg/d). The patients were followed up for 5 years. Baseline mean arterial pressures, which were 109 +/- 3 mm Hg in the amlodipine group and 108 +/- 3 mm Hg in the enalapril group, decreased significantly after 1 year of follow-up (amlodipine, 96 +/- 3 mm Hg; P < 0.005; enalapril, 89 +/- 2 mm Hg; P < 0.0005) and remained stable at year 5 (amlodipine, 97 +/- 3 mm Hg; P < 0.0005 versus baseline; enalapril, 94 +/- 3 mm Hg; P < 0.005 versus baseline). Ccrs, which were 83 +/- 5 mL/min/1.73 m(2) in the amlodipine group and 77 +/- 6 mL/min/1.73 m(2) in the enalapril group, remained stable after 1 year of follow up and decreased significantly at year 3 in both groups (amlodipine, 67 +/- 5 mL/min/1.73 m(2); P < 0.01 versus year 1 and baseline; enalapril, 58 +/- 4 mL/min/1.73 m(2); P < 0.05 versus year 1 and P < 0.0005 versus baseline) with no significant change thereafter. No change was observed in urinary albumin creatinine ratio in the amlodipine group (baseline, 68 +/- 21 mg/g; year 1, 52 +/ 21 mg/g; year 5, 148 +/- 74 mg/g), whereas it decreased significantly in the enalapril group at year 1 (baseline, 23 +/- 4 mg/g; year 1, 13 +/- 3 mg/g; P < 0.05) and remained stable until the end of the study at year 5 (14 +/- 6 mg/g). The investigators concluded that blood pressure was similar in both groups but only enalapril had a significant effect to sustain decreased urinary albumin excretion for a 5-year follow-up. Although proteinuria has been considered a surrogate of renal disease progression, further studies will be necessary to confirm this hypothesis in ADPKD, because after 5 years, no differences in renal function were observed between the enalapril and amlodipine groups. In comparison with patients with ADPKD with uncontrolled hypertension, effective control of blood pressure, as undertaken in the present study, should delay the onset of ESRD by approximately 15 years. PMID- 10692269 TI - Etiologies and outcome of acute renal insufficiency in older adults: a renal biopsy study of 259 cases. AB - Acute renal insufficiency is a common problem, yet one that is frequently reversible with proper diagnosis and treatment. Although it has been argued that a renal biopsy is not needed for diagnosis in most cases of acute renal failure in the elderly, other studies have shown frequent disagreements between clinical and renal biopsy diagnoses in such cases. To investigate the causes of acute renal insufficiency in patients aged at least 60 years who underwent a renal biopsy and possible correlations between biopsy findings and renal survival, we first identified all native renal biopsy specimens from patients aged 60 years or older processed at The University of Chicago Medical Center (Chicago, IL) from 1991 through 1998 and reviewed the clinical records to determine the indication for the biopsy. We then reviewed again the records of those patients who underwent biopsy because of acute renal insufficiency, recorded the primary renal biopsy diagnosis in each of these cases, and obtained follow-up information for patients who underwent biopsy before July 1996. During the study period, 1,065 of 4,264 biopsy specimens (25.0%) received were obtained from patients aged 60 years or older, and acute renal insufficiency was the indication for biopsy in 259 of these patients (24.3%). The most frequent primary diagnoses on these latter biopsy specimens were pauci-immune crescentic glomerulonephritis (GN) with or without arteritis, 31.2% of biopsy specimens; acute interstitial nephritis, 18.6%; acute tubular necrosis (ATN) with nephrotic syndrome, 7.5%; atheroemboli, 7.1%; ATN alone, 6.7%; light chain cast nephropathy (LCCN), 5.9%; postinfectious GN, 5.5%; anti-glomerular basement membrane antibody nephritis, 4.0%; and immunoglobulin A (IgA) nephropathy and/or Henoch-Schonlein nephritis, 3.6%. Eight biopsy specimens (3.2%) showed only benign nephrosclerosis without an apparent cause of acute renal insufficiency, and another six specimens were inadequate. The renal biopsy diagnosis was in agreement with the prebiopsy clinical diagnosis (or differential diagnosis) in 107 of the 161 cases (67%) in which such information was provided. The distribution of diagnoses was similar in patients in the age groups of 60 to 69, 70 to 79, and 80 years or older, although younger age correlated significantly with improved renal and patient survival. The relative risk for progression to end-stage renal disease (ESRD) also increased according to diagnostic categories: LCCN (greatest risk) > GN other than pauci immune > atheroemboli congruent with pauci-immune crescentic GN > tubulointerstitial diseases other than LCCN (the latter category including ATN with nephrotic syndrome). Development of ESRD correlated significantly with decreased patient survival. In summary, renal biopsy in patients aged 60 years or older with acute renal insufficiency uncovered the cause in greater than 90% of the cases and provided clinically useful information with respect to expectation for renal survival and potential treatment options. PMID- 10692270 TI - Current indications for renal biopsy: a questionnaire-based survey. AB - Indications for renal biopsy are still ill defined. We recently sent a detailed questionnaire to 360 nephrologists in different areas of the world with the aim of providing information on this critical issue by evaluating the replies. The questionnaire was organized in four sections that included questions on renal biopsy indications in patients with normal renal function, renal insufficiency, and a transplanted kidney. In addition, the questions included methods applied to each renal biopsy procedure and to specimen processing. We received 166 replies; North Europe (50 replies), South Europe (47 replies), North America (31 replies), Australia and New Zealand (24 replies), and other countries (14 replies). In patients with normal renal function, primary indications for renal biopsy were microhematuria associated with proteinuria, particularly greater than 1 g/d of protein. In chronic renal insufficiency, kidney dimension was the major parameter considered before renal biopsy, whereas the presence of diabetes or serological abnormalities was not considered critical. In the course of acute renal failure (ARF) of unknown origin, 20% of the respondents would perform renal biopsy in the early stages, 26% after 1 week of nonrecovery, and 40% after 4 weeks. In a transplanted kidney, the majority of nephrologists would perform a renal biopsy in the case of graft failure after surgery, ARF after initial good function, slow progressive deterioration of renal function, and onset of nephrotic proteinuria. The last section provided comprehensive information on the technical aspects of renal biopsy. This survey represents the first attempt to provide a reliable consensus that can be used in developing guidelines on the use of kidney biopsy. PMID- 10692271 TI - Effect of 22-oxacalcitriol on bone histology of hemodialyzed patients with severe secondary hyperparathyroidism. AB - To examine the effectiveness of 22-oxacalcitriol (OCT) injection on the improvement of severe osteitis fibrosa, we studied 10 hemodialyzed patients (age, 59 +/- 12 years). The initial OCT dose was 5 microg and was administered three times weekly at the end of each hemodialysis session. OCT doses (1, 3, 5, 10, 15, and 20 microg) were changed in subsequent weeks to maintain serum calcium levels at less than 11.5 mg/dL. Administration of OCT significantly suppressed serum intact parathyroid hormone (PTH) from an initial level of 1,193 +/- 584 to 775 +/ 552 pg/mL in the 24th week (n = 10). OCT increased PTH levels again to 857 +/- 635 pg/mL in the 48th week (n = 7). Among the 10 patients, 5 patients (high responders) showed more than a 50% suppression of serum intact PTH levels at the end of the study. The rest of the patients had hypercalcemia and did not receive increased OCT doses (low responders). At the start of the treatment, the only difference between high and low responders was serum calcium level. Serum calcium levels (adjusted for serum albumin level) increased from 9.7 +/- 0.7 mg/dL (n = 10) at the beginning to 10.5 +/- 0.6 mg/dL (n = 10) in the 24th week and to 11. 1 +/- 0.7 mg/dL (n = 7) in the 48th week. Six patients (1 to 6) agreed to undergo a second bone biopsy in the 24th week of OCT administration. In bone histomorphometric measurements, OCT significantly changed bone marrow fibrosis, mineralization (labeled mineralizing surface and bone formation rate), and osteoid formation (osteoid volume and thickness). In conclusion, intravenous OCT effectively suppressed PTH secretion and improved the bone histological characteristics of severe osteitis fibrosa, especially in patients with initial serum calcium levels less than 10 mg/dL. With concerns about OCT causing adynamic bone, additional bone histological data are needed to ensure the long-term safety of OCT. PMID- 10692272 TI - Presence of sonographically detectable parathyroid glands can predict resistance to oral pulsed-dose calcitriol treatment of secondary hyperparathyroidism. AB - Oral pulsed-dose calcitriol administration has been shown to be effective therapy for patients with secondary hyperparathyroidism. However, this effect is not consistently observed in the clinical setting. This study was undertaken to examine whether enlarged parathyroid glands can serve as a clinical marker that predicts the suppressive effect of calcitriol. Thirty-five patients undergoing chronic hemodialysis (HD) were examined (age, 51.9 +/- 14.9 years; duration of HD, 72.0 +/- 56.0 months). Based on the volume of parathyroid glands measured using an ultrasonographic scanner, patients were divided into two groups: 15 patients with undetectable parathyroid glands (ND group) and 20 patients with detectable parathyroid glands (D group; mean volume of parathyroid glands, 261. 9 +/- 347.5 mm(3)). No significant differences were found in serum ionized calcium (Ca(++)) or parathyroid hormone (PTH) levels before calcitriol administration between the two groups. For each patient, 8 microg of calcitriol was administered orally at the end of the HD session, and the changes in serum PTH levels were determined 44 hours after dosing. No significant differences were found between the two groups in serum Ca(++) levels. However, a significant decrease in serum PTH levels was observed in the ND group, whereas no significant changes were found in the D group. The results of the study show that the suppressive effect on PTH through calcitriol therapy was reduced in patients with detectable enlarged parathyroid glands. This may indicate that the size of parathyroid glands is one factor determining the therapeutic potential of pulsed-dose calcitriol administration for secondary hyperparathyroidism. PMID- 10692273 TI - C-Reactive protein predicts all-cause and cardiovascular mortality in hemodialysis patients. AB - Hypoalbuminemia predicts death in dialysis patients. Although hypoalbuminemia has been attributed to malnutrition, evidence of inflammation (C-reactive protein [CRP] and cytokine levels) has recently been recognized to predict albumin concentration in dialysis patients. We measured CRP and albumin levels in October 1995 in 91 hemodialysis (HD) patients. During a 34-month follow-up period, we determined the incidence and cause of death. Patients were divided into four groups based on serum albumin levels (<3.5 [lowest quartile], 3.5 to 3.8, 3.9 to 4.0, and >4.0 g/dL [highest quartile]). Survival differed among the four groups (P = 0.0063). Patients with albumin levels greater than 4.0 g/dL had the greatest survival. Kaplan-Meier survival estimates of patients from varying CRP quartiles (<2.6, 2.6 to 5.2, 5.3 to 11.5, and >11.5 microg/mL) differed among the four groups (P < 0.0001). The group with the greatest CRP level (>11.5 microg/mL) had the lowest survival. Multivariate analysis using the Cox proportional hazards model showed that only CRP level (chi-square = 21.11; P < 0.0001) and age (chi square = 5.44; P = 0.020) predicted death. Albumin level (chi-square = 0.16; P = 0.69) was not predictive. Only when CRP was excluded from the model did low serum albumin level (chi-square = 12. 04; P = 0.0004) predict death. CRP level (chi square = 16.79; P < 0. 0001) and age (chi-square = 6.38; P = 0.012) also superceded albumin level (chi-square = 0.45; P = 0.51) in predicting cardiovascular mortality. Although values for blood urea nitrogen, creatinine, and normalized protein catabolic rate were significantly less among patients who died, these parameters, as well as cholesterol level and diabetes, were not important predictors of death in multivariate analysis. The acute-phase response or the cause of the acute-phase response is largely responsible for the effect of hypoalbuminemia on mortality in HD patients. PMID- 10692274 TI - New ultrasound approaches to dialysis access monitoring. AB - The failure of dialysis access is a frequent source of morbidity and hospitalization. Traditional methods of graft surveillance include: (1) clinical examination, (2) venous line pressure measurements during dialysis, (3) urea or tracer recirculation measurement, (4) continuous wave (CW) Doppler methods, (5) duplex ultrasonography, and (6) radiograph angiography. All these methods require special training and/or laboratory tests. The purpose of this study was to test a simple continuous-wave Doppler method that could be applied to measure the flow rate in dialysis access every time the patient undergoes dialysis. Twenty dialysis patients, 15 with polytetrafluoroethylene grafts and 5 with arteriovenous fistulae, were studied. Two hundred fifty-three examinations were performed over an 8-month period. Doppler waveforms of the access flow were obtained with the pump on, with the pump off, and with the pump on again. Systolic and diastolic Doppler frequency measurements were made, and the pump-on and pump-off measurements were compared. In an access functioning normally, the Doppler frequencies are higher with the pump off than with the pump on. In 22% of the cases, there were abnormal findings in which the Doppler frequencies were lower with the pump off than with the pump on. This occurs if the needles are incorrectly placed, suggesting that recirculation is occurring. Recirculation also occurs if there is stenosis of the access. Examining the hemodialysis access during each dialysis session with an inexpensive directional Doppler may identify a significant stenosis and improve the efficiency of dialysis by detecting those patients in whom the arterial and venous needles are reversed. PMID- 10692275 TI - Physical functioning and health-related quality-of-life changes with exercise training in hemodialysis patients. AB - The Renal Exercise Demonstration Project was designed to test the effects of two different approaches to exercise programming on the levels of physical activity, physical functioning, and self-reported health status in hemodialysis patients. Two hundred eighty-six patients were recruited for participation. Intervention patients were given individually prescribed exercise for 8 weeks of independent home exercise, followed by 8 weeks of incenter cycling during dialysis. Physical performance testing was performed at baseline and after each intervention using gait speed, sit-to-stand test, and 6-minute walk. The Medical Outcomes Study Short Form 36-item (SF-36) questionnaire was used to assess self-reported health status. The intervention group showed increased participation in physical activity. There were significant differences between the intervention and nonintervention groups in change over time in normal and fast gait speed, sit-to stand test scores, and the physical scales on the SF-36, including the physical component scale. The intervention group improved in these test results, whereas the nonintervention group either did not change or declined over the duration of the study. It is clear that improvements in physical functioning result from exercise counseling and encouragement in hemodialysis patients. Because self reported physical functioning is highly predictive of outcomes in hemodialysis patients, more attention to patients' levels of physical activity is warranted. PMID- 10692277 TI - Prolonged sulfonylurea-induced hypoglycemia in diabetic patients with end-stage renal disease. AB - Renal impairment is a recognized risk factor for prolonged hypoglycemia, but predisposing characteristics in patients with advanced renal impairment have not been studied. We observed prolonged hypoglycemia in a number of patients with end stage renal disease (ESRD) and conducted a case-control study at two Canadian centers to identify such risk factors. Through hospital, pharmacy, and dialysis program records, we retrospectively identified 7 case patients and 31 controls with ESRD and type 2 diabetes using oral hypoglycemic monotherapy. Control patients had no history of hospital admission for prolonged hypoglycemia. All case patients and 28 controls were receiving glyburide (glibenclamide in Europe); the remainder were treated with tolbutamide. Duration of intravenous treatment for hypoglycemia ranged from 28 to 256 hours, with 83 g to 2 kg of glucose administered per episode. Preceding treatment with glyburide varied from 2 days to 13 years. Univariate analyses showed a recent decline in oral intake (odds ratio [OR], 81; 95% confidence interval [CI], 3.6 to 1,840), previous hypoglycemic episodes (OR, 15; 95% CI, 0.77 to 297), longer duration of diabetes (22 versus 12 years; P = 0.008), and a history of cerebrovascular disease (OR, 7. 0; 95% CI, 1.0 to 47) to be associated with prolonged hypoglycemia. No association between prolonged hypoglycemia and age, sex, beta blockers, angiotensin-converting enzyme inhibitors, oral hypoglycemic dose, or duration of treatment was identified. This study describes the potentially devastating effect of sulfonylurea-based oral hypoglycemic therapy in ESRD. Patients at greatest risk appear to be those with reduced intake, previous hypoglycemic episodes, and longer duration of diabetes. We describe the mechanisms for observed hypoglycemia and suggest that alternative drugs may be considered in this patient group. PMID- 10692276 TI - Dialysate made from dry chemicals using citric acid increases dialysis dose. AB - A new dry dialysate concentrate acidified with citric acid (citrate dialysate) has been used in two separate clinical studies of hemodialysis patients. The first compared a single treatment using this dialysate, with one dialysis using regular standard dialysate acidified with acetic acid (regular dialysate) in a prospective, randomized, crossover study of 74 dialyses. Changes in blood levels of electrolytes and other blood constituents during dialysis were calculated by subtracting postdialysis from predialysis blood concentrations. Compared with acetic acid dialysate, citrate dialysate was associated with significantly greater decreases in total and ionized calcium, magnesium, and chloride levels. Citrate dialysate was also associated with greater increases in serum sodium and citrate concentrations, although their postdialysis concentrations remained within or just outside normal ranges. Changes in other blood constituents were similar with both dialysates. The second study used citrate dialysate exclusively for all dialyses over a 12-week period in 25 patients. Predialysis blood samples were drawn at the start of the study and at 4-week intervals thereafter, and postdialysis blood samples were obtained after the first and last dialysis. Repeated-measure analysis showed that although predialysis blood concentrations of magnesium, potassium, and citrate remained within the normal range, there was a significant declining trend over the course of the study. At the same time, predialysis serum bicarbonate levels increased, and significantly more patients had a predialysis bicarbonate concentration within the normal range at the end of the study than at the start (15 versus 8 patients; P = 0.001, chi-square). In 19 patients (excluding 3 patients for whom the type of dialyzer was changed during the study), the dose of dialysis for the first and last dialysis was calculated by urea reduction ratio and Kt/V. There was a significant increase in both measurements without changes in dialysis time, blood and dialysate flows, or dialyzer used. The urea reduction ratio increased from 68% +/- 5.9% to 73% +/- 5.3% (P < 0. 03), and the Kt/V from 1.23 +/- 0.19 to 1.34 +/- 0.20 (P = 0.01) from the first to last dialysis, respectively. In conclusion, this citric acid dialysate was well tolerated, and intradialytic changes in blood chemistries were similar to those seen with regular dialysate. Using dialysate containing citric instead of acetic acid increases the delivered dialysis dose. PMID- 10692278 TI - A multicenter study of noncompliance with continuous ambulatory peritoneal dialysis exchanges in US and Canadian patients. AB - Recent evidence suggested that noncompliance (NC) with continuous ambulatory peritoneal dialysis (CAPD) exchanges may be more common in US than in Canadian dialysis centers. This issue was investigated using a questionnaire-based method in 656 CAPD patients at 14 centers in the United States and Canada. NC was defined as missing more than one exchange per week or more than two exchanges per month. Patients were ensured of the confidentiality of their individual results. Mean patient age was 56 +/- 16 years, 52% were women, and 39% had diabetes. The overall admitted rate of NC was 13%, with a rate of 18% in the United States and 7% in Canada (P < 0.001). NC was more common in younger patients (P < 0.0001), those without diabetes (P < 0.001), and employed patients (P < 0.05). It was also more common in black and Hispanic than in Asian and white patients (P < 0.001). NC was more common in patients prescribed more than four exchanges daily (P < 0.0001) but was not affected by dwell volume. On multiple regression analysis, the independent predictors of NC, in order of importance, were being prescribed more than four exchanges per day, black race, being employed, younger age, and not having diabetes. Being treated in a US unit did not quite achieve significance as a multivariate independent predictor. These findings suggest that NC is not uncommon in CAPD patients and is more frequent in US than in Canadian patients. However, country of residence is less powerful as a predictor of NC than a variety of other demographic and prescription factors. PMID- 10692279 TI - Multicenter cross-sectional study for dialysis dose and physician's subjective judgment in Japanese peritoneal dialysis patients. Group for the Water and Electrocyte Balance Study in CAPD. AB - The purpose of this study was to investigate the state of dialysis and nutrition among Japanese peritoneal dialysis (PD) patients. Two hundred thirty-nine Japanese PD patients from 21 centers, 79 female and 160 male, were evaluated to determine their status of dialysis and nutrition. Mean age of the patients was 50 years; mean duration on PD, 4.2 years; mean body weight, 58 kg; and body surface area (BSA), 1.61 m(2). Sixty-three percent of the patients had no residual renal function. Mean daily delivered volume was 6.9 L for female continuous ambulatory peritoneal dialysis (CAPD), 8.1 L for male CAPD, 10.5 L for female automated peritoneal dialysis (APD), and 10.7 L for male APD. Total (dialysis and kidney) mean weekly values for creatinine clearance (Ccr), Kt/V, and beta2 microglobulin (beta2MG) clearance were 56 L/1.75 m(2), 1.80, and 11 L/1.73 m(2), respectively. Fifty percent of the patients who had no residual renal function (RRF) and 17% of the patients with RRF did not achieve 50 L/wk/1.73 m(2) of Ccr. With regard to nutritional parameters, mean values for plasma total protein, serum albumin, and normalized protein catabolic rate (nPCR) were 6.5 g/dL, 3.6 g/dL, and 0.97 g/kg BW/d. Mean daily protein loss was 5.8 g. Although a significant number of patients did not achieve 50 or 60 L/wk/1.73 m(2) of Ccr, the physicians determined that 72% of the patients received adequate dialysis and 71% were well nourished according to clinical and laboratory features. In conclusion, the daily prescribed volume and the delivered dialysis dose were lower than expected. The discrepancy between the actual delivered dialysis dose and the physicians' evaluation should be explored in the future. PMID- 10692280 TI - Ultrasound detection of microembolic signals in hemodialysis accesses. AB - Microembolic signals (MES) detected by ultrasound, thought to be of gaseous or solid origin, have been described with decompression illness and in the intracranial and cardiopulmonary circulation. We describe the first reported cases of MES occurring in hemodialysis accesses. Two hemodialysis patients, one with a synthetic graft and one with an arteriovenous fistula, showed MES during a dialysis session detected by duplex ultrasound. We postulate that these MES represent cavitation bubbles developing from turbulent blood flow around the venous needle in the access. However, other potential causes exist, including air introduced into the circulation from the dialysis circuit or microemboli arising from thrombus or atheroma. PMID- 10692281 TI - Access recirculation in a native fistula in spite of a seemingly adequate access flow. AB - True access recirculation (AR) measured by ultrasound dilution technique is usually absent in well-working shunts. It occurs with low access flows (Qa). High access flow rates are assumed to prevent AR. Two major exceptions to these rules are known: presence of intra-access strictures and inadvertently reversed blood lines. We present an additional exception in which true access recirculation occurred in a native arteriovenous (AV) fistula with correct placement of bloodlines. Surprisingly, access blood flow exceeded pump blood flow (Qb) almost threefold. The situation was clarified by a magnetic resonance angiogram showing a collateral forming a functional loop. This loop led to true access recirculation in one branch, although overall blood flow through both branches appeared to be adequate. The different findings in this shunt over time give insight into the often complex pathophysiology of native fistulae. This case proves that seemingly adequate access flow does not necessarily prevent access recirculation in native AV fistulae. We suggest monitoring both access flow and recirculation in hemodialysis accesses on a regular basis. PMID- 10692282 TI - Thin basement membrane disease and acute renal failure secondary to gross hematuria and tubular necrosis. AB - A patient with thin basement membrane disease (TBMD), macroscopic hematuria, and acute renal failure is described. A renal biopsy showed massive occlusion of renal tubules by red blood cells and casts. This was accompanied by tubular cell damage consistent with acute tubular necrosis. The patient was receiving warfarin because of a history of deep venous thrombosis at the time he developed the acute renal failure. The possible relationship of the warfarin therapy to the TBMD, intratubular hemorrhage, and acute renal failure are discussed. PMID- 10692283 TI - Thrombocytopenia after kidney transplantation. AB - We report a case of posttransplant malaria in which the patient developed progressive thrombocytopenia after receiving living related donor kidney transplantation. The donor, who flew in from Sri Lanka for the procedure, had suffered from malaria 18 months earlier. Malaria should be suspected in transplant patients receiving organs from donors originating from countries with a high prevalence of malaria. PMID- 10692284 TI - Long-term cyclophosphamide treatment for recurrent type I membranoproliferative glomerulonephritis after transplantation. AB - The incidence of recurrent type I membranoproliferative glomerulonephritis (MPGN) after renal transplant is approximately 30%, and the rate of graft loss due to recurrent MPGN type I is higher than 50%. The treatment of this disease has not been defined. We report a case of recurrent MPGN type diagnosed 4 months after a cadaveric renal transplantation. The patient was treated with cyclophosphamide and was able to maintain her graft function. Cyclophosphamide was interrupted three times during the course. Each time her renal function deteriorated and her serum albumin decreased. The patient currently has a functional renal graft 3 years after transplantation while receiving low-dose therapy with cyclophosphamide. We suggest treating recurrent type I MPGN with cyclophosphamide while continuing the calcineurin inhibitor and prednisone. PMID- 10692285 TI - Renal biopsy in the elderly. PMID- 10692286 TI - Hypertension, proteinuria, and progression of autosomal dominant polycystic kidney disease: where do we go from here? PMID- 10692287 TI - IgA1 glycosylation and the pathogenesis of IgA nephropathy. PMID- 10692288 TI - Acute renal failure with interstitial nephritis in a patient with AIDS. PMID- 10692289 TI - Risk for posttransplant diabetes mellitus with current immunosuppressive medications. PMID- 10692290 TI - Hypertension in the hemodialysis patient. PMID- 10692294 TI - Disappearance of nodular mesangial lesions in a patient with light chain nephropathy after long-term chemotherapy. AB - A 64-year-old man developed multiple myeloma (kappa light chain type), nephrotic syndrome, and renal insufficiency in 1993. A renal biopsy showed typical histological findings of light chain nephropathy: nodular glomerulosclerosis with deposition of kappa light chains in the mesangial area and subendothelial space of the glomerular capillary walls. Long-term intermittent MEVP chemotherapy (melphalan, 4 mg/d for 4 days; cyclophosphamide, 100 mg/d for 4 days; vincristine, 1 mg/d; prednisolone, 40 mg/d for 4 days) diminished proteinuria and improved renal function. In April 1999, a follow-up biopsy showed remarkable diminution of nodular lesions and disappearance of kappa light chain deposits. Although the prognosis of light chain nephropathy has been considered poor, long term successful chemotherapy can clear light chain deposits and restore renal function. PMID- 10692295 TI - De novo ANCA-associated vasculitis occurring 14 years after kidney transplantation. AB - A cadaveric kidney transplant recipient, with no history of a connective tissue disease, was admitted with malaise, arthralgias, diplopia, mild headache, and a painful left eye. The patient was on maintenance immunosuppression for 14 years with cyclosporine and methylprednisolone. Initial laboratory data indicated an elevated serum creatinine from baseline, 2+ proteinuria, and 50 to 100 red blood cells (RBCs)/high-power field (HPF) in the urine. Renal biopsy was consistent with necrotizing vasculitis involving glomerular capillaries, with crescent formation and an absence of immune complexes. Perinuclear antineutrophil cytoplasmic autoantibodies (P-ANCA) and anti-myeloperoxidase (MPO) were found to be elevated. To the best of our knowledge, this is the first reported case of an ANCA-associated small vessel vasculitis (SVV) developing in a renal transplant recipient without history of connective tissue disease. PMID- 10692296 TI - Extensive intraglomerular thrombi of monoclonal IgM-kappa in a patient with malignant lymphoma. AB - We describe an 80-year-old man who developed malignant lymphoma (ML) complicated by extensive intraglomerular thrombi of immunoglobulin M (IgM)-kappa monoclonal immunoglobulin. The clinical picture was characterized by nephrotic syndrome and systemic lymphadenopathy. Laboratory examination showed mild anemia and a small amount of monoclonal IgM-kappa in the blood. The histopathologic findings and surface immunoglobulin analysis of the lymph node biopsy specimen were consistent with CD5-positive diffuse large B-cell (type, IgM-kappa) lymphoma. The subsequent renal biopsy showed a massive deposition of amorphous material in the glomerular capillary lumens, subendothelial areas, and mesangium. Nodular glomerulosclerosis was not found. An immunofluorescent study showed that the deposits consisted of IgM-kappa monoclonal immunoglobulin. Ultrastructurally, the deposits were composed of granular electron-dense material. Chemotherapy was effective for both the ML and nephrotic syndrome, and the patient's urine analysis results returned to normal. The histopathologic manifestations of this case are rare, and the pathogenesis of these glomerular lesions was obviously associated with ML. PMID- 10692297 TI - Reactivation of tuberculosis after conversion from azathioprine to mycophenolate mofetil 16 years after renal transplantation. AB - The incidence of tuberculosis among transplant recipients is greater than in the general population. Mycophenolate mofetil (MMF) is a potent immunosuppressive agent that has become part of most standard immunosuppressive protocols after renal transplantation. We have recently shown that conversion from azathioprine (AZA) to MMF in patients with chronic allograft dysfunction may be beneficial. Here, we report a patient with a history of pulmonary tuberculosis during his childhood. This patient was converted from AZA to MMF therapy 16 years after allogenic renal transplantation because of chronic allograft dysfunction. Two months later, he developed axillary lymph node tuberculosis caused by Mycobacterium tuberculosis. Because he denied contact with infectious persons, we diagnosed reactivation of old dormant tuberculosis. After surgical extirpation, quadruple antituberculous therapy was administered for 3 months (isoniazid, rifampicin, ethambutol, and pyrazinamide), followed by dual therapy for 3 months (isoniazid and rifampicin), and monotherapy for another 3 months (isoniazid). In the follow-up period, he remained asymptomatic with stable graft function. We conclude that MMF therapy in renal allograft recipients may cause reactivation of old dormant tuberculosis, even in the very late posttransplantation period. In these patients, close monitoring and isoniazid prophylaxis may be useful. PMID- 10692298 TI - Physiologic and pathophysiologic renal consequences of H(+)-stimulated endothelin secretion. PMID- 10692299 TI - How yeast cells synchronize their glycolytic oscillations: a perturbation analytic treatment. AB - Of all the lifeforms that obtain their energy from glycolysis, yeast cells are among the most basic. Under certain conditions the concentrations of the glycolytic intermediates in yeast cells can oscillate. Individual yeast cells in a suspension can synchronize their oscillations to get in phase with each other. Although the glycolytic oscillations originate in the upper part of the glycolytic chain, the signaling agent in this synchronization appears to be acetaldehyde, a membrane-permeating metabolite at the bottom of the anaerobic part of the glycolytic chain. Here we address the issue of how a metabolite remote from the pacemaking origin of the oscillation may nevertheless control the synchronization. We present a quantitative model for glycolytic oscillations and their synchronization in terms of chemical kinetics. We show that, in essence, the common acetaldehyde concentration can be modeled as a small perturbation on the "pacemaker" whose effect on the period of the oscillations of cells in the same suspension is indeed such that a synchronization develops. PMID- 10692300 TI - Kinetics of desolvation-mediated protein-protein binding. AB - The role of desolvation in protein binding kinetics is investigated using Brownian dynamics simulations in complexes in which the electrostatic interactions are relatively weak. We find that partial desolvation, modeled by a short-range atomic contact potential, is not only a major contributor to the binding free energy but also substantially increases the diffusion-limited rate for complexes in which long-range electrostatics is weak. This rate enhancement is mostly due to weakly specific pathways leading to a low free-energy attractor, i.e., a precursor state before docking. For alpha-chymotrypsin and human leukocyte elastase, both interacting with turkey ovomucoid third domain, we find that the forward rate constant associated with a collision within a solid angle phi around their corresponding attractor approaches 10(7) and 10(6) M(-1)s(-1), respectively, in the limit phi approximately 2 degrees. Because these estimates agree well with experiments, we conclude that the final bound conformation must be preceded by a small set of well-defined diffusion-accessible precursor states. The inclusion of the otherwise repulsive desolvation interaction also explains the lack of aggregation in proteins by restricting nonspecific association times to approximately 4 ns. Under the same reaction conditions but without short range forces, the association rate would be only approximately 10(3) M(-1)s(-1). Although desolvation increases these rates by three orders of magnitude, desolvation-mediated association is still at least 100-fold slower than the electrostatically assisted binding in complexes such as barnase and barstar. PMID- 10692301 TI - Quantal potential fields around individual active zones of amphibian motor-nerve terminals. AB - The release of a quantum from a nerve terminal is accompanied by the flow of extracellular current, which creates a field around the site of transmitter action. We provide a solution for the extent of this field for the case of a quantum released from a site on an amphibian motor-nerve terminal branch onto the receptor patch of a muscle fiber and compare this with measurements of the field using three extracellular electrodes. Numerical solution of the equations for the quantal potential field in cylindrical coordinates show that the density of the field at the peak of the quantal current gives rise to a peak extracellular potential, which declines approximately as the inverse of the distance from the source at distances greater than about 4 microm from the source along the length of the fiber. The peak extracellular potential declines to 20% of its initial value in a distance of about 6 microm, both along the length of the fiber and in the circumferential direction around the fiber. Simultaneous recordings of quantal potential fields, made with three electrodes placed in a line at right angles to an FM1-43 visualized branch, gave determinations of the field strengths in accord with the numerical solutions. In addition, the three electrodes were placed so as to straddle the visualized release sites of a branch. The positions of these sites were correctly predicted on the basis of the theory and independently ascertained by FM1-43 staining of the sites. It is concluded that quantal potential fields at the neuromuscular junction that can be measured with available recording techniques are restricted to regions within about 10 microm of the release site. PMID- 10692302 TI - Effect of voltage drop within the synaptic cleft on the current and voltage generated at a single synapse. AB - In a model of a single synapse with a circular contact zone and a single concentric zone containing receptor-gated channels, we studied the dependence of the synaptic current on the synaptic cleft width and on the relative size of the receptor zone. During synaptic excitation, the extracellular current entered the cleft and flowed into the postsynaptic cell through receptor channels distributed homogeneously over the receptor zone. The membrane potential and channel currents were smaller toward the cleft center if compared to the cleft edges. This radial gradient was due to the voltage drop produced by the synaptic current on the cleft resistance. The total synaptic current conducted by the same number of open channels was sensitive to changes in the receptor zone radius and the cleft width. We conclude that synaptic geometry may affect synaptic currents by defining the volume resistor of the cleft. The in-series connection of the resistances of the intracleft medium and the receptor channels plays the role of the synaptic voltage divider. This voltage dividing effect should be taken into account when the conductance of single channels or synaptic contacts is estimated from experimental measurements of voltage-current relationships. PMID- 10692303 TI - Backbone dipoles generate positive potentials in all proteins: origins and implications of the effect. AB - Asymmetry in packing the peptide amide dipole results in larger positive than negative regions in proteins of all folding motifs. The average side chain potential in 305 proteins is 109 +/- 30 mV (2. 5 +/- 0.7 kcal/mol/e). Because the backbone has zero net charge, the non-zero potential is unexpected. The larger oxygen at the negative and smaller proton at the positive end of the amide dipole yield positive potentials because: 1) at allowed phi and psi angles residues come off the backbone into the positive end of their own amide dipole, avoiding the large oxygen; and 2) amide dipoles with their carbonyl oxygen surface exposed and amine proton buried make the protein interior more positive. Twice as many amides have their oxygens exposed than their amine protons. The distribution of acidic and basic residues shows the importance of the bias toward positive backbone potentials. Thirty percent of the Asp, Glu, Lys, and Arg are buried. Sixty percent of buried residues are acids, only 40% bases. The positive backbone potential stabilizes ionization of 20% of the acids by >3 pH units (-4.1 kcal/mol). Only 6.5% of the bases are equivalently stabilized by negative regions. The backbone stabilizes bound anions such as phosphates and rarely stabilizes bound cations. PMID- 10692304 TI - Transduction of intracellular and intercellular dynamics in yeast glycolytic oscillations. AB - Under certain well-defined conditions, a population of yeast cells exhibits glycolytic oscillations that synchronize through intercellular acetaldehyde. This implies that the dynamic phenomenon of the oscillation propagates within and between cells. We here develop a method to establish by which route dynamics propagate through a biological reaction network. Application of the method to yeast demonstrates how the oscillations and the synchronization signal can be transduced. That transduction is not so much through the backbone of glycolysis, as via the Gibbs energy and redox coenzyme couples (ATP/ADP, and NADH/NAD), and via both intra- and intercellular acetaldehyde. PMID- 10692305 TI - A model for the lipid pretransition: coupling of ripple formation with the chain melting transition. AB - Below the thermotropic chain-melting transition, lipid membrane c(P) traces display a transition of low enthalpy called the lipid pretransition. It is linked to the formation of periodic membrane ripples. In the literature, these two transitions are usually regarded as independent events. Here, we present a model that is based on the assumption that both pretransition and main transition are caused by the same physical effect, namely chain melting. The splitting of the melting process into two peaks is found to be a consequence of the coupling of structural changes and chain-melting events. On the basis of this concept, we performed Monte Carlo simulations using two coupled monolayer lattices. In this calculation, ripples are considered to be one-dimensional defects of fluid lipid molecules. Because lipids change their area by approximately 24% upon melting, line defects are the only ones that are topologically possible in a triangular lattice. The formation of a fluid line defect on one monolayer leads to a local bending of the membrane. Geometric constraints result in the formation of periodic patterns of gel and fluid domains. This model, for the first time, is able to predict heat capacity profiles, which are comparable to the experimental c(P) traces that we obtained using calorimetry. The basic assumptions are in agreement with a large number of experimental observations. PMID- 10692306 TI - Properties of the stochastic energization-relaxation channel model for vectorial ion transport. AB - A model for the primary active transport by an ion pump protein is proposed. The model, the "energization-relaxation channel model," describes an ion pump as a multiion channel that undergoes stochastic transitions between two conformational states by external energy supply. When the potential profile along ion transport pathway is asymmetrical, a net ion flux is induced by the transitions. In this model, the coupling of the conformational change and ion transport is stochastic and loose. The model qualitatively reproduces known properties of active transport such as the effect of ion concentration gradient and membrane potential on the rate of transport and the inhibition of ion transport at high ion concentration. We further examined the effect of various parameters on the ion transport properties of this model. The efficiency of the coupling was almost 100% under some conditions. PMID- 10692307 TI - Mechanism of lateral movement of filopodia and radial actin bundles across neuronal growth cones. AB - We investigated the motion of filopodia and actin bundles in lamellipodia of motile cells, using time-lapse sequences of polarized light images. We measured the velocity of retrograde flow of the actin network and the lateral motion of filopodia and actin bundles of the lamellipodium. Upon noting that laterally moving filopodia and actin bundles are always tilted with respect to the direction of retrograde flow, we propose a simple geometric model for the mechanism of lateral motion. The model establishes a relationship between the speed of lateral motion of actin bundles, their tilt angle with respect to the direction of retrograde flow, and the speed of retrograde flow in the lamellipodium. Our experimental results verify the quantitative predictions of the model. Furthermore, our observations support the hypothesis that lateral movement of filopodia is caused by retrograde flow of tilted actin bundles and by their growth through actin polymerization at the tip of the bundles inside the filopodia. Therefore we conclude that the lateral motion of tilted filopodia and actin bundles does not require a separate motile mechanism but is the result of retrograde flow and the assembly of actin filaments and bundles near the leading edge of the lamellipodium. PMID- 10692308 TI - Rigidity of triskelion arms and clathrin nets. AB - Statistical analysis is applied to a set of electron micrographic images (Kocsis, E., B. L. Trus, C. J. Steer, M. E. Bisher, and A. C. Steven. 1991. J. Struct. Biol. 107:6-14), from which quantitative measures are obtained to support the notion that the three arms of a triskelion have statistically identical properties and exhibit independent structural fluctuations. Additionally, a study of local contour fluctuations, which indicates that the elastic properties of a triskelion arm are approximately constant over the entire arm length, is used along with a small deformation statistical mechanics theory to derive an effective, average flexural rigidity for the arms. This result is used to estimate the bending energy necessary to deform a clathrin patch, and comparison is made with the deformation energy of an equivalent area of non-clathrin-coated membrane. We estimate that the rigidity of the clathrin lattice is at least comparable to that of a typical membrane. Hence, the natural curvature of a clathrin cage can stabilize, and perhaps propel, the formation of intracellular coated vesicles. PMID- 10692309 TI - Time and force dependence of the rupture of glycoprotein IIb-IIIa-fibrinogen bonds between latex spheres. AB - We studied the shear-induced breakup of doublets of aldehyde/sulfate (A/S) latex spheres covalently linked with purified platelet GPIIb-IIIa receptor, and cross linked by fibrinogen. Flow cytometry with fluorescein isothiocyanate-fibrinogen showed than an average of 22,500 molecules of active GPIIb-IIIa were captured per sphere, with a mean K(d) = 56 nM for fibrinogen binding. The spheres, suspended in buffered 19% Ficoll 400 containing 120 or 240 pM fibrinogen, were subjected to Couette flow in a counter-rotating cone-plate rheoscope. Doublets, formed by two body collisions at low shear rate (G = 8 s(-1)) for < or =15 min, were subjected to shear stress from 0.6 to 2.9 Nm(-2), their rotations recorded until they broke up or were lost to view. Although breakup was time dependent, occurring mostly in the first 2 rotations after the onset of shear, the percentage of doublets broken up after 10 rotations were almost independent of normal hydrodynamic force, F(n): at 240 pN, 15.6, 16.0, and 17.0% broke up in the force range 70-150 pN, 150-230 pN, and 230-310 pN. Unexpectedly, at both [fibrinogen], the initial rate of breakup was highest in the lowest force range, and computer simulation using a stochastic model of breakup was unable to simulate the time course of breakup. When pre-sheared at low G for >15 min, no doublets broke up within 10 rotations at 70 < F(n) < 310 pN; it required >3 min shear (>1110 rotations) at F(n) = 210 pN for significant breakup to occur. Other published work has shown that binding of fibrinogen to GPIIb-IIIa immobilized on plane surfaces exhibits an initial fast reversible process with relative low affinity succeeded by transformation of GPIIb-IIIa to a stable high-affinity complex. We postulate that most doublet breakups observed within 10 rotations were from a population of young doublets having low numbers of bonds, by dissociation of the initial receptor complex relatively unresponsive to force. The remaining, older doublets with GPIIb-IIIa in the high-affinity complex were not broken up in the time or range of forces studied. PMID- 10692310 TI - Response kinetics of tethered Rhodobacter sphaeroides to changes in light intensity. AB - Rhodobacter sphaeroides can swim toward a wide range of attractants (a process known as taxis), propelled by a single rotating flagellum. The reversals of motor direction that cause tumbles in Eschericia coli taxis are replaced by brief motor stops, and taxis is controlled by a complex sensory system with multiple homologues of the E. coli sensory proteins. We tethered photosynthetically grown cells of R. sphaeroides by their flagella and measured the response of the flagellar motor to changes in light intensity. The unstimulated bias (probability of not being stopped) was significantly larger than the bias of tethered E. coli but similar to the probability of not tumbling in swimming E. coli. Otherwise, the step and impulse responses were the same as those of tethered E. coli to chemical attractants. This indicates that the single motor and multiple sensory signaling pathways in R. sphaeroides generate the same swimming response as several motors and a single pathway in E. coli, and that the response of the single motor is directly observable in the swimming pattern. Photo-responses were larger in the presence of cyanide or the uncoupler carbonyl cyanide 4 trifluoromethoxyphenylhydrazone (FCCP), consistent with the photo-response being detected via changes in the rate of electron transport. PMID- 10692311 TI - Direct observation of trapping and release of nitric oxide by glutathione and cysteine with electron paramagnetic resonance spectroscopy. AB - While the biosynthesis of nitric oxide (NO) is well established, one of the key issues that remains to be solved is whether NO participates in the biological responses right after generation through biosynthesis or there is a "secret passage" via which NO itself is trapped, transported, and released to exert its functions. It has been shown that NO reacts with thiol-containing biomolecules (RSH), like cysteine (Cys), glutathione (GSH), etc., to form S-nitrosothiols (RSNOs), which then release nitrogen compounds, including NO. The direct observation of trapping of NO and its release by RSNO has not been well documented, as most of the detection techniques measure the content of NO as well as nitrite and nitrate. Here we use spin-trapping electron paramagnetic resonance (EPR) technique to measure NO content directly in the reaction time course of samples of GSH and Cys ( approximately mM) mixed with NO ( approximately microM) in the presence of metal ion chelator, which pertains to physiological conditions. We demonstrate that NO is readily trapped by these thiols in less than 10 min and approximately 70-90% is released afterward. These data imply that approximately 10-30% of the reaction product of NO does not exist in the free radical form. The NO release versus time curves are slightly pH dependent in the presence of metal ion chelator. Because GSH and Cys exist in high molar concentrations in blood and in mammalian cells, the trapping and release passage of NO by these thiols may provide a mechanism for temporal and spatial sequestration of NO to overcome its concentration gradient-dependent diffusion, so as to exert its multiple biological effects by reacting with various targets through regeneration. PMID- 10692312 TI - Coexpression of alpha and beta subunits of the rod cyclic GMP-gated channel restores native sensitivity to cyclic AMP: role of D604/N1201. AB - Coexpression of the betawt and alphawt subunits of the bovine rod channel restores two characteristics of the native channels: higher sensitivity to cAMP and potentiation of cGMP-induced currents by low cAMP concentrations. To test whether the increased sensitivity to cAMP is due to the uncharged nature of the asparagine residue (N1201) situated in place of aspartate D604 in the beta subunit as previously suggested (, Neuron. 15:619-625), we compared currents from wild-type (alphawt and alphawt/betawt) and from mutated channels (alphaD604N, alphaD604N/betawt, and alphawt/betaN1201D). The results show that the sensitivity to cAMP and cAMP potentiation is partly but not entirely determined by the charge of residue 1201 in the beta subunit. The D604N mutation in the alpha subunit and, to a lesser extent, coexpression of the betawt subunit with the alphawt subunit reduce the open probability for cGMP compared to that of the alphawt channel. Interpretation of the data with the MWC allosteric model (model of Monod, Wyman, Changeux;, J. Mol. Biol. 12:88-118) suggests that the D604N mutation in the alpha subunits and coassembly of alpha and beta subunits alter the free energy of gating by cAMP more than that of cAMP binding. PMID- 10692313 TI - Calcium currents in hair cells isolated from semicircular canals of the frog. AB - L-type and R-type Ca(2+) currents were detected in frog semicircular canal hair cells. The former was noninactivating and nifedipine-sensitive (5 microM); the latter, partially inactivated, was resistant to omega-conotoxin GVIA (5 microM), omega-conotoxin MVIIC (5 microM), and omega-agatoxin IVA (0.4 microM), but was sensitive to mibefradil (10 microM). Both currents were sensitive to Ni(2+) and Cd(2+) (>10 microM). In some cells the L-type current amplitude increased almost twofold upon repetitive stimulation, whereas the R-type current remained unaffected. Eventually, run-down occurred for both currents, but was prevented by the protease inhibitor calpastatin. The R-type current peak component ran down first, without changing its plateau, suggesting that two channel types generate the R-type current. This peak component appeared at -40 mV, reached a maximal value at -30 mV, and became undetectable for voltages > or =0 mV, suggestive of a novel transient current: its inactivation was indeed reversibly removed when Ba(2+) was the charge carrier. The L-type current and the R-type current plateau were appreciable at -60 mV and peaked at -20 mV: the former current did not reverse for voltages up to +60 mV, the latter reversed between +30 and +60 mV due to an outward Cs(+) current flowing through the same Ca(2+) channel. The physiological role of these currents on hair cell function is discussed. PMID- 10692314 TI - Histidine(118) in the S2-S3 linker specifically controls activation of the KAT1 channel expressed in Xenopus oocytes. AB - The guard cell K(+) channel KAT1, cloned from Arabidopsis thaliana, is activated by hyperpolarization and regulated by a variety of physiological factors. Low internal pH accelerated the activation kinetics of the KAT1 channel expressed in Xenopus oocytes with a pK of approximately 6, similar to guard cells in vivo. Mutations of histidine-118 located in the putative cytoplasmic linker between the S2 and S3 segments profoundly affected the gating behavior and pH dependence. At pH 7.2, substitution with a negatively charged amino acid (glutamate, aspartate) specifically slowed the activation time course, whereas that with a positively charged amino acid (lysine, arginine) accelerated. These mutations did not alter the channel's deactivation time course or the gating behavior after the first opening. Introducing an uncharged amino acid (alanine, asparagine) at position 118 did not have any obvious effect on the activation kinetics at pH 7.2. The charged substitutions markedly decreased the sensitivity of the KAT1 channel to internal pH in the physiological range. We propose a linear kinetic scheme to account for the KAT1 activation time course at the voltages where the opening transitions dominate. Changes in one forward rate constant in the model adequately account for the effects of the mutations at position 118 in the S2-S3 linker segment. These results provide a molecular and biophysical basis for the diversity in the activation kinetics of inward rectifiers among different plant species. PMID- 10692315 TI - Molecular cloning of cDNA encoding a drosophila ryanodine receptor and functional studies of the carboxyl-terminal calcium release channel. AB - Ryanodine is a plant alkaloid that was originally used as an insecticide. To study the function and regulation of the ryanodine receptor (RyR) from insect cells, we have cloned the entire cDNA sequence of RyR from the fruit fly Drosophila melanogaster. The primary sequence of the Drosophila RyR contains 5134 amino acids, which shares approximately 45% identity with RyRs from mammalian cells, with a large cytoplasmic domain at the amino-terminal end and a small transmembrane domain at the carboxyl-terminal end. To characterize the Ca(2+) release channel activity of the cloned Drosophila RyR, we expressed both full length and a deletion mutant of Drosophila RyR lacking amino acids 277-3650 (Drosophila RyR-C) in Chinese hamster ovary cells. For subcellular localization of the expressed Drosophila RyR and Drosophila RyR-C proteins, green fluorescent protein (GFP)-Drosophila RyR and GFP-Drosophila RyR-C fusion constructs were generated. Confocal microscopic imaging identified GFP-Drosophila RyR and GFP Drosophila RyR-C on the endoplasmic reticulum membranes of transfected cells. Upon reconstitution into the lipid bilayer membrane, Drosophila RyR-C formed a large conductance cation-selective channel, which was sensitive to modulation by ryanodine. Opening of the Drosophila RyR-C channel required the presence of microM concentration of Ca(2+) in the cytosolic solution, but the channel was insensitive to inhibition by Ca(2+) at concentrations as high as 20 mM. Our data are consistent with our previous observation with the mammalian RyR that the conduction pore of the calcium release channel resides within the carboxyl terminal end of the protein and further demonstrate that structural and functional features are essentially shared by mammalian and insect RyRs. PMID- 10692316 TI - Functional expression of the L-type calcium channel in mice skeletal muscle during prenatal myogenesis. AB - The densities of skeletal muscle intramembrane charge movement and macroscopic L type Ca(2+) current have been shown to increase during prenatal development. In the present work, the electrophysiological characteristics of L-type Ca(2+) channels were analyzed over the embryonic period E14 to E19 using the whole-cell and cell-attached procedures. At the macroscopic level, the whole-cell L-type Ca(2+) conductance increased 100% between E14 and E19. This enhancement was accompanied by a small negative shift of the voltage dependence and a marked acceleration of the inactivation kinetics. At the single-channel level, the unitary conductance decreased significantly from 13.2 +/- 0.1 pS (n = 8) at E14 to 10.7 +/- 0.3 pS (n = 7) at E18 and the open probability was multiplied by 2. No significant change of the density of functional channels was observed during the same period. In contrast to the density of intramembrane charge movement, which, under the same conditions, has been shown to increase between 16 and 19 days, L-type Ca(2+) channels properties change mostly between 14 and 16 days. Taken together, these results suggest that the two functions of the dihydropyridine receptor are carried by two different proteins which could be differentially regulated by subunit composition and/or degree of phosphorylation. PMID- 10692317 TI - Conformation, independent of charge, in the R domain affects cystic fibrosis transmembrane conductance regulator channel openings. AB - The R domain of cystic fibrosis transmembrane conductance regulator (CFTR), when phosphorylated, undergoes conformational change, and the chloride channel opens. We investigated the contribution of R domain conformation, apart from the changes induced by phosphorylation, to channel opening, by testing the effect of the peptidyl-prolyl isomerase, cyclophilin A, on the CFTR channel. When it was applied after the channel had been opened by PKA phosphorylation, cyclophilin A increased the open probability of wild-type CFTR (from P(o) = 0.197 +/- 0.010 to P(o) = 0.436 +/- 0. 029) by increasing the number of channel openings, not open time. Three highly conserved proline residues in the R domain, at positions 740, 750, and 759, were considered as candidate targets for cyclophilin A. Mutations of these prolines to alanines (P3A mutant) resulted in a channel unresponsive to cyclophilin A but with pore properties similar to the wild type, under strict control of PKA and ATP, but with significantly increased open probability (P(o) = 0.577 +/- 0.090) compared to wild-type CFTR, again due to an increase in the number of channel openings and not open time. Mutation of each of the proline residues separately and in pairs demonstrated that all three proline mutations are required for maximal P(o). When P3A was expressed in 293 HEK cells and tested by SPQ assay, chloride efflux was significantly increased compared to cells transfected with wild-type CFTR. Thus, treatments favoring the trans-peptidyl conformation about conserved proline residues in the R domain of CFTR affect openings of CFTR, above and beyond the effect of PKA phosphorylation. PMID- 10692318 TI - Kinetics studies of the cardiac Ca-ATPase expressed in Sf21 cells: new insights on Ca-ATPase regulation by phospholamban. AB - Kinetics studies of the cardiac Ca-ATPase expressed in Sf21 cells (Spodoptera frugiperda insect cells) have been carried out to test the hypotheses that phospholamban inhibits Ca-ATPase cycling by decreasing the rate of the E1.Ca to E1'.Ca transition and/or the rate of phosphoenzyme hydrolysis. Three sample types were studied: Ca-ATPase expressed alone, Ca-ATPase coexpressed with wild-type phospholamban (the natural pentameric inhibitor), and Ca-ATPase coexpressed with the L37A-phospholamban mutant (a more potent monomeric inhibitor, in which Leu(37) is replaced by Ala). Phospholamban coupling to the Ca-ATPase was controlled using a monoclonal antibody against phospholamban. Gel electrophoresis and immunoblotting confirmed an equivalent ratio of Ca-ATPase and phospholamban in each sample (1 mol Ca-ATPase to 1.5 mol phospholamban). Steady-state ATPase activity assays at 37 degrees C, using 5 mM MgATP, showed that the phospholamban containing samples had nearly equivalent maximum activity ( approximately 0.75 micromol. nmol Ca-ATPase(-1).min(-1) at 15 microM Ca(2+)), but that wild-type phospholamban and L37A-phospholamban increased the Ca-ATPase K(Ca) values by 200 nM and 400 nM, respectively. When steady-state Ca-ATPase phosphoenzyme levels were measured at 0 degrees C, using 1 microM MgATP, the K(Ca) values also shifted by 200 nM and 400 nM, respectively, similar to the results obtained by measuring ATP hydrolysis at 37 degrees C. Measurements of the time course of phosphoenzyme formation at 0 degrees C, using 1 microM MgATP and 268 nM ionized [Ca(2+)], indicated that L37A-phospholamban decreased the steady-state phosphoenzyme level to a greater extent (45%) than did wild-type phospholamban (33%), but neither wild-type nor L37A-phospholamban had any effect on the apparent rate of phosphoenzyme formation relative to that of Ca-ATPase expressed alone. Measurements of inorganic phosphate (P(i)) release concomitant with the phosphoenzyme formation studies showed that L37A-phospholamban decreased the steady-state rate of P(i) release to a greater extent (45%) than did wild-type phospholamban (33%). However, independent measurements of Ca-ATPase dephosphorylation after the addition of 5 mM EGTA to the phosphorylated enzyme showed that neither wild-type phospholamban nor L37A-phospholamban had any effect on the rate of phosphoenzyme decay relative to Ca-ATPase expressed alone. Computer simulation of the kinetics data indicated that phospholamban and L37A phospholamban decreased twofold and fourfold, respectively, the equilibrium binding of the first Ca(2+) ion to the Ca-ATPase E1 intermediate, rather than inhibiting rate of the E.Ca to E'.Ca transition or the rate of phosphoenzyme decay. Therefore, we conclude that phospholamban inhibits Ca-ATPase cycling by decreasing Ca-ATPase Ca(2+) binding to the E1 intermediate. PMID- 10692319 TI - Electrostatic interactions regulate desensitization of the nicotinic acetylcholine receptor. AB - To determine the importance of electrostatic interactions for agonist binding to the nicotinic acetylcholine receptor (AChR), we examined the affinity of the fluorescent agonist dansyl-C6-choline for the AChR. Increasing ionic strength decreased the binding affinity in a noncompetitive manner and increased the Hill coefficient of binding. Small cations did not compete directly for dansyl-C6 choline binding. The sensitivity to ionic strength was reduced in the presence of proadifen, a noncompetitive antagonist that desensitizes the receptor. Moreover, at low ionic strength, the dansyl-C6-choline affinities were similar in the absence or presence of proadifen, a result consistent with the receptor being desensitized at low ionic strength. Similar ionic strength effects were observed for the binding of the noncompetitive antagonist [(3)H]ethidium when examined in the presence and absence of agonist to desensitize the AChR. Therefore, ionic strength modulates binding affinity through at least two mechanisms: by influencing the conformation of the AChR and by electrostatic effects at the binding sites. The results show that charge-charge interactions regulate the desensitization of the receptor. Analysis of dansyl-C6-choline binding to the desensitized conformation using the Debye-Huckel equation was consistent with the presence of five to nine negative charges within 20 A of the acetylcholine binding sites. PMID- 10692320 TI - Protonation of lysine residues inverts cation/anion selectivity in a model channel. AB - A dimeric alamethicin analog with lysine at position 18 in the sequence (alm-K18) was previously shown to form stable anion-selective channels in membranes at pH 7.0 [Starostin, A. V., R. Butan, V. Borisenko, D. A. James, H. Wenschuh, M. S. Sansom, and G. A. Woolley. 1999. Biochemistry. 38:6144-6150]. To probe the charge state of the conducting channel and how this might influence cation versus anion selectivity, we performed a series of single-channel selectivity measurements at different pH values. At pH 7.0 and below, only anion-selective channels were found with P(K(+))/P(Cl(-)) = 0. 25. From pH 8-10, a mixture of anion-selective, non-selective, and cation-selective channels was found. At pH > 11 only cation selective channels were found with P(K(+))/P(Cl(-)) = 4. In contrast, native alamethicin-Q18 channels (with Gln in place of Lys at position 18) were cation selective (P(K(+))/P(Cl(-)) = 4) at all pH values. Continuum electrostatics calculations were then carried out using an octameric model of the alm-K18 channel embedded in a low dielectric slab to simulate a membrane. Although the calculations can account for the apparent pK(a) of the channel, they fail to correctly predict the degree of selectivity. Although a switch from cation- to anion-selectivity as the channel becomes protonated is indicated, the degree of anion-selectivity is severely overestimated, suggesting that the continuum approach does not adequately represent some aspect of the electrostatics of permeation in these channels. Side-chain conformational changes upon protonation, conformational changes, and deprotonation caused by permeating cations and counterion binding by lysine residues upon protonation are considered as possible sources of the overestimation. PMID- 10692321 TI - Amino acid residues 4425-4621 localized on the three-dimensional structure of the skeletal muscle ryanodine receptor. AB - We have localized a region contained within the sequence of amino acid residues 4425-4621 on the three-dimensional structure of the skeletal muscle ryanodine receptor (RyR). Mouse monoclonal antibodies raised against a peptide comprising these residues have been complexed with ryanodine receptors and imaged in the frozen-hydrated state by cryoelectron microscopy. These images, along with images of antibody-free ryanodine receptor, were used to compute two-dimensional averaged images and three-dimensional reconstructions. Two-dimensional averages of immunocomplexes in which the ryanodine receptor was in the fourfold symmetrical orientation disclosed four symmetrical regions of density located on the edges of the receptor's cytoplasmic assembly that were absent from control averages of receptor without added antibody. Three-dimensional reconstructions revealed the antibody-binding sites to be on the so-called handle domains of the ryanodine receptor's cytoplasmic assembly, near their junction with the transmembrane assembly. This study is the first to demonstrate epitope mapping on the three-dimensional structure of the ryanodine receptor. PMID- 10692322 TI - Protein-induced membrane disorder: a molecular dynamics study of melittin in a dipalmitoylphosphatidylcholine bilayer. AB - A molecular dynamics simulation of melittin in a hydrated dipalmitoylphosphatidylcholine (DPPC) bilayer was performed. The 19, 000-atom system included a 72-DPPC phospholipid bilayer, a 26-amino acid peptide, and more than 3000 water molecules. The N-terminus of the peptide was protonated and embedded in the membrane in a transbilayer orientation perpendicular to the surface. The simulation results show that the peptide affects the lower (intracellular) layer of the bilayer more strongly than the upper (extracellular) layer. The simulation results can be interpreted as indicating an increased level of disorder and structural deformation for lower-layer phospholipids in the immediate vicinity of the peptide. This conclusion is supported by the calculated deuterium order parameters, the observed deformation at the intracellular interface, and an increase in fractional free volume. The upper layer was less affected by the embedded peptide, except for an acquired tilt relative to the bilayer normal. The effect of melittin on the surrounding membrane is localized to its immediate vicinity, and its asymmetry with respect to the two layers may result from the fact that it is not fully transmembranal. Melittin's hydrophilic C-terminus anchors it at the extracellular interface, leaving the N-terminus "loose" in the lower layer of the membrane. In general, the simulation supports a role for local deformation and water penetration in melittin-induced lysis. As for the peptide, like other membrane-embedded polypeptides, melittin adopts a significant 25 degree tilt relative to the membrane normal. This tilt is correlated with a comparable tilt of the lipids in the upper membrane layer. The peptide itself retains an overall helical structure throughout the simulation (with the exception of the three N-terminal residues), adopting a 30 degree intrahelical bend angle. PMID- 10692323 TI - Cholesterol effects on the phosphatidylcholine bilayer polar region: a molecular simulation study. AB - A molecular dynamics (MD) simulation of a fully hydrated, liquid-crystalline dimyristoylphosphatidylcholine (DMPC)-Chol bilayer membrane containing approximately 22 mol% Chol was carried out for 4.3 ns. The bilayer reached thermal equilibrium after 2.3 ns of MD simulation. A 2.0-ns trajectory generated during 2.3-4.3 ns of MD simulation was used for analyses to determine the effects of Chol on the membrane/water interfacial region. In this region, 70% of Chol molecules are linked to DMPC molecules via short-distance interactions, where the Chol hydroxyl group (OH-Chol) is 1) charge paired to methyl groups of the DMPC choline moiety ( approximately 34%), via the hydroxyl oxygen atom (Och); 2) water bridged to carbonyl ( approximately 19%) and nonester phosphate ( approximately 14%) oxygen atoms, via both Och and the hydroxyl hydrogen atom (Hch); and 3) directly hydrogen (H) bonded to carbonyl ( approximately 11%) and nonester phosphate ( approximately 5%) oxygen atoms, via Hch ( approximately 17% of DMPC Chol links are multiple). DMPC's gamma-chain carbonyl oxygen atom is involved in 44% of water bridges and 51% of direct H bonds formed between DMPC and Chol. On average, a Chol molecule forms 0.9 links with DMPC molecules, while a DMPC molecule forms 2.2 and 0.3 links with DMPC and Chol molecules, respectively. OH Chol makes hydrogen bonds with 1.1 water molecules, preferentially via Hch. The average number of water molecules H bonded to the DMPC headgroup is increased by 7% in the presence of Chol. These results indicate that inclusion of Chol decreases interlipid links and increases hydration in the polar region of the membrane. PMID- 10692324 TI - Studies of archaebacterial bipolar tetraether liposomes by perylene fluorescence. AB - Membrane packing and dynamics of bipolar tetraether liposomes composed of the polar lipid fraction E (PLFE) from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius have been studied by perylene fluorescence. At a probe to-PLFE lipid ratio of 1:400, we have detected an unusual fluorescence intensity increase with increasing temperature, while the fluorescence lifetime changed little. As the ratio was decreased, the intensity anomaly was diminished. At 1:3200 and 1:6400, the anomaly disappeared. A remarkable perylene intensity anomaly was also observed in bilayers composed of saturated monopolar diester phosphatidylcholines at their main phase transition temperatures. These results suggest that the intensity anomaly may be due to probe aggregation caused by tight membrane packing. At the same probe-to-lipid ratio (1:400), however, 1, 2 diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and 1, 2-diphytanoyl-sn-glycero-3 phosphoglycerol (DPhPG) liposomes did not exhibit any intensity anomaly with increasing temperature. This suggests that DPhPC and DPhPG liposomes are more loosely packed than PLFE liposomes; thus the branched methyl groups are not the contributing factor of the tight membrane packing found in PLFE liposomes. Using a multiexcitation method, we have also determined the average (R), in-plane (R(ip)), and out-of-plane (R(op)) rotational rates of perylene in PLFE liposomes at various temperatures (20-65 degrees C). R and R(ip), determined at two different probe-to-lipid ratios (1:400 and 1:3200), both undergo an abrupt increase when the temperature is elevated to approximately 48 degrees C. These data suggest that PLFE liposomes are rigid and tightly packed at low temperatures, but they begin to possess appreciable "membrane fluidity" at temperatures close to the minimum growth temperature ( approximately 50 degrees C) of thermoacidophilic archaebacteria. PMID- 10692325 TI - Interaction of phosphatidylserine synthase from E. coli with lipid bilayers: coupled plasmon-waveguide resonance spectroscopy studies. AB - The interaction of phosphatidylserine (PS) synthase from Escherichia coli with lipid membranes was studied with a recently developed variant of the surface plasmon resonance technique, referred to as coupled plasmon-waveguide resonance spectroscopy. The features of the new technique are increased sensitivity and spectral resolution, and a unique ability to directly measure the structural anisotropy of lipid and proteolipid films. Solid-supported lipid bilayers with the following compositions were used: 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC); POPC-1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate (POPA) (80:20, mol/mol); POPC-POPA (60:40, mol/mol); and POPC-1-palmitoyl-2-oleoyl-sn glycero-3-[phospho-rac-(1-glycerol)] (POPG) (75:25, mol/mol). Addition of either POPA or POPG to a POPC bilayer causes a considerable increase of both the bilayer thickness and its optical anisotropy. PS synthase exhibits a biphasic interaction with the bilayers. The first phase, occurring at low protein concentrations, involves both electrostatic and hydrophobic interactions, although it is dominated by the latter, and the enzyme causes a local decrease of the ordering of the lipid molecules. The second phase, occurring at high protein concentrations, is predominantly controlled by electrostatic interactions, and results in a cooperative binding of the enzyme to the membrane surface. Addition of the anionic lipids to a POPC bilayer causes a 5- to 15-fold decrease in the protein concentration at which the first binding phase occurs. The results reported herein lend experimental support to a previously suggested mechanism for the regulation of the polar head group composition in E. coli membranes. PMID- 10692326 TI - Kinetic and structural study of the interaction of myelin basic protein with dipalmitoylphosphatidylglycerol layers. AB - The interaction of myelin basic protein (MBP) with dipalmitoylphosphatidylglycerol films has been investigated by means of a microgravimetric gauge sensitive to the changes in load and structural modifications of the layer deposited onto its surface. Fourier transform infrared spectroscopy, circular dichroism, and x-ray diffraction have confirmed protein uptake by the lipid phase along with a global disordering effect onto the lipid alkyl chains and have shown a temporal evolution of the structure of water penetrating the lipid phase together with the protein. These effects are clearly related to the temporal variation of the microgravimetric gauge signal. Finally, measurements carried out on pre-annealed samples point out the role of mesoscopic morphology in determining the pathways through which MBP penetrates the lipid multilayer. The results obtained in our model system could be useful in clarifying the mechanisms of the myelinating and demyelinating processes that take place in the natural membrane. PMID- 10692327 TI - Molecular and mesoscopic properties of hydrophilic polymer-grafted phospholipids mixed with phosphatidylcholine in aqueous dispersion: interaction of dipalmitoyl N-poly(ethylene glycol)phosphatidylethanolamine with dipalmitoylphosphatidylcholine studied by spectrophotometry and spin-label electron spin resonance. AB - Spin-label electron spin resonance (ESR) spectroscopy, together with optical density measurements, has been used to investigate, at both the molecular and supramolecular levels, the interactions of N-poly(ethylene glycol) phosphatidylethanolamines (PEG-PE) with phosphatidylcholine (PC) in aqueous dispersions. PEG-PEs are micelle-forming hydrophilic polymer-grafted lipids that are used extensively for steric stabilization of PC liposomes to increase their lifetimes in the blood circulation. All lipids had dipalmitoyl (C16:0) chains, and the polymer polar group of the PEG-PE lipids had a mean molecular mass of either 350 or 2000 Da. PC/PEG-PE mixtures were investigated over the entire range of relative compositions. Spin-label ESR was used quantitatively to investigate bilayer-micelle conversion with increasing PEG-PE content by measurements at temperatures for which the bilayer membrane component of the mixture was in the gel phase. Both saturation transfer ESR and optical density measurements were used to obtain information on the dependence of lipid aggregate size on PEG-PE content. It is found that the stable state of lipid aggregation is strongly dependent not only on PEG-PE content but also on the size of the hydrophilic polar group. These biophysical properties may be used for optimized design of sterically stabilized liposomes. PMID- 10692328 TI - The effect of removing the N-terminal extension of the Drosophila myosin regulatory light chain upon flight ability and the contractile dynamics of indirect flight muscle. AB - The Drosophila myosin regulatory light chain (DMLC2) is homologous to MLC2s of vertebrate organisms, except for the presence of a unique 46-amino acid N terminal extension. To study the role of the DMLC2 N-terminal extension in Drosophila flight muscle, we constructed a truncated form of the Dmlc2 gene lacking amino acids 2-46 (Dmlc2(Delta2-46)). The mutant gene was expressed in vivo, with no wild-type Dmlc2 gene expression, via P-element-mediated germline transformation. Expression of the truncated DMLC2 rescues the recessive lethality and dominant flightless phenotype of the Dmlc2 null, with no discernible effect on indirect flight muscle (IFM) sarcomere assembly. Homozygous Dmlc2(Delta2-46) flies have reduced IFM dynamic stiffness and elastic modulus at the frequency of maximum power output. The viscous modulus, a measure of the fly's ability to perform oscillatory work, was not significantly affected in Dmlc2(Delta2-46) IFM. In vivo flight performance measurements of Dmlc2(Delta2-46) flies using a visual closed-loop flight arena show deficits in maximum metabolic power (P(*)(CO(2))), mechanical power (P(*)(mech)), and flight force. However, mutant flies were capable of generating flight force levels comparable to body weight, thus enabling them to fly, albeit with diminished performance. The reduction in elastic modulus in Dmlc2(Delta2-46) skinned fibers is consistent with the N terminal extension being a link between the thick and thin filaments that is parallel to the cross-bridges. Removal of this parallel link causes an unfavorable shift in the resonant properties of the flight system, thus leading to attenuated flight performance. PMID- 10692329 TI - Dynamics at Lys-553 of the acto-myosin interface in the weakly and strongly bound states. AB - Lys-553 of skeletal muscle myosin subfragment 1 (S1) was specifically labeled with the fluorescent probe FHS (6-[fluorescein-5(and 6)-carboxamido]hexanoic acid succinimidyl ester) and fluorescence quenching experiments were carried out to determine the accessibility of this probe at Lys-553 in both the strongly and weakly actin-bound states of the MgATPase cycle. Solvent quenchers of varying charge [nitromethane, (2,2,6, 6-tetramethyl-1-piperinyloxy) (TEMPO), iodide (I( )), and thallium (Tl(+))] were used to assess both the steric and electrostatic accessibilities of the FHS probe at Lys-553. In the strongly bound rigor (nucleotide-free) and MgADP states, actin offered no protection from solvent quenching of FHS by nitromethane, TEMPO, or thallium, but did decrease the Stern Volmer constant by almost a factor of two when iodide was used as the quencher. The protection from iodide quenching was almost fully reversed with the addition of 150 mM KCl, suggesting this effect is ionic in nature rather than steric. Conversely, actin offered no protection from iodide quenching at low ionic strength during steady-state ATP hydrolysis, even with a significant fraction of the myosin heads bound to actin. Thus, the lower 50 kD subdomain of myosin containing Lys-553 appears to interact differently with actin in the weakly and strongly bound states. PMID- 10692330 TI - Detection of fluorescently labeled actin-bound cross-bridges in actively contracting myofibrils. AB - Myosin subfragment 1 (S1) can be specifically modified at Lys-553 with the fluorescent probe FHS (6-[fluorescein-5(and 6)-carboxamido]hexanoic acid succinimidyl ester) (Bertrand, R., J. Derancourt, and R. Kassab. 1995. Biochemistry. 34:9500-9507), and solvent quenching of FHS-S1 with iodide has been shown to be sensitive to actin binding at low ionic strength (MacLean, Chrin, and Berger, 2000. Biophys. J. 000-000). In order to extend these results and examine the fraction of actin-bound myosin heads within the myofilament lattice during calcium activation, we have modified skeletal muscle myofibrils, mildly cross linked with EDC (1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide) to prevent shortening, with FHS. The myosin heavy chain appears to be the predominant site of labeling, and the iodide quenching patterns are consistent with those obtained for myosin S1 in solution, suggesting that Lys-553 is indeed the primary site of FHS incorporation in skeletal muscle myofibrils. The iodide quenching results from calcium-activated FHS-myofibrils indicate that during isometric contraction 29% of the myosin heads are strongly bound to actin within the myofilament lattice at low ionic strength. These results suggest that myosin can be specifically modified with FHS in more complex and physiologically relevant preparations, allowing the real time examination of cross-bridge interactions with actin in in vitro motility assays and during isometric and isotonic contractions within single muscle fibers. PMID- 10692331 TI - Cross-bridge attachment during high-speed active shortening of skinned fibers of the rabbit psoas muscle: implications for cross-bridge action during maximum velocity of filament sliding. AB - To characterize the kinetics of cross-bridge attachment to actin during unloaded contraction (maximum velocity of filament sliding), ramp-shaped stretches with different stretch-velocities (2-40,000 nm per half-sarcomere per s) were applied to actively contracting skinned fibers of the rabbit psoas muscle. Apparent fiber stiffness observed during such stretches was plotted versus the speed of the imposed stretch (stiffness-speed relation) to derive the rate constants for cross bridge dissociation from actin. The stiffness-speed relation obtained for unloaded shortening conditions was shifted by about two orders of magnitude to faster stretch velocities compared to isometric conditions and was almost identical to the stiffness-speed relation observed in the presence of MgATPgammaS at high Ca(2+) concentrations, i.e., under conditions where cross-bridges are weakly attached to the fully Ca(2+) activated thin filaments. These data together with several control experiments suggest that, in contrast to previous assumptions, most of the fiber stiffness observed during high-speed shortening results from weak cross-bridge attachment to actin. The fraction of strongly attached cross-bridges during unloaded shortening appears to be as low as some 1 5% of the fraction present during isometric contraction. This is about an order of magnitude less than previous estimates in which contribution of weak cross bridge attachment to observed fiber stiffness was not considered. Our findings imply that 1) the interaction distance of strongly attached cross-bridges during high-speed shortening is well within the range consistent with conventional cross bridge models, i.e., that no repetitive power strokes need to be assumed, and 2) that a significant part of the negative forces that limit the maximum speed of filament sliding might originate from weak cross-bridge interactions with actin. PMID- 10692332 TI - CaATP as a substrate to investigate the myosin lever arm hypothesis of force generation. AB - In an effort to test the lever arm model of force generation, the effects of replacing magnesium with calcium as the ATP-chelated divalent cation were determined for several myosin and actomyosin reactions. The isometric force produced by glycerinated muscle fibers when CaATP is the substrate is 20% of the value obtained with MgATP. For myosin subfragment 1 (S1), the degree of lever arm rotation, determined using transient electric birefringence to measure rates of rotational Brownian motion in solution, is not significantly changed when calcium replaces magnesium in an S1-ADP-vanadate complex. Actin activates S1 CaATPase activity, although less than it does MgATPase activity. The increase in actin affinity when S1. CaADP. P(i) is converted to S1. CaADP is somewhat greater than it is for the magnesium case. The ionic strength dependence of actin binding indicates that the change in apparent electrostatic charge at the acto-S1 interface for the S1. CaADP. P(i) to S1. CaADP step is similar to the change when magnesium is bound. In general, CaATP is an inferior substrate compared to MgATP, but all the data are consistent with force production by a lever arm mechanism for both substrates. Possible reasons for the reduced magnitude of force when CaATP is the substrate are discussed. PMID- 10692333 TI - Structural implications of the chemical modification of Cys(10) on actin. AB - Cys(10) is located in subdomain 1 of actin, which has an important role in the interaction of actin with myosin- and actin-binding proteins. Cys(10) was modified with fluorescence probes N-(iodoacetyl)N'-(5-sulfo-1-naphthyl)ethylene diamine (IAEDANS), 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM), or monobromo bimane (MBB) by the method of, J. Biol. Chem. 266:5508-5513). The specificity of Cys(10) modification was verified by showing that the 33-kDa subtilisin fragment of actin (residues 48-375), which contains all of the actin thiols but Cys(10), is not fluorescent. Cys(10) modification exposed a new site on actin to subtilisin cleavage. Edman degradation revealed this site to be between Ala(19) and Gly(20). The modification slightly increased the rate of epsilonATP-ATP exchange and decreased the rates of G-actin ATPase and polymerization. The activation of S1 ATPase by Cys(10)-modified F-actin showed small probe-dependent changes in the values of V(max) and K(M). The sliding speed of actin filaments in the in vitro motility assay remained unchanged upon modification of Cys(10). These results indicate that although the labeling of Cys(10) perturbs the structure of subdomain 1, the modified actin remains fully functional. The binding of S1 to actin filaments decreases the accessibility of Cys(10) probes to acrylamide and nitromethane quenchers. Because Cys(10) does not participate directly in either actin polymerization or S1 binding, our results indicate that actin-actin and actin-myosin interactions induce dynamic, allosteric changes in actin structure. PMID- 10692334 TI - Morphology and transverse stiffness of Drosophila myofibrils measured by atomic force microscopy. AB - Atomic force microscopy was used to investigate the surface morphology and transverse stiffness of myofibrils from Drosophila indirect flight muscle exposed to different physiologic solutions. I- and A-bands were clearly observed, and thick filaments were resolved along the periphery of the myofibril. Interfilament spacings correlated well with estimates from previous x-ray diffraction studies. Transverse stiffness was measured by using a blunt tip to indent a small section of the myofibrillar surface in the region of myofilament overlap. At 10 nm indention, the effective transverse stiffness (K( perpendicular)) of myofibrils in rigor solution (ATP-free, pCa 4.5) was 10.3 +/- 5.0 pN nm(-1) (mean +/- SEM, n = 8); in activating solution (pCa 4.5), 5.9 +/- 3.1 pN nm(-1); and in relaxing solution (pCa 8), 4.4 +/- 2.0 pN nm(-1). The apparent transverse Young's modulus (E( perpendicular)) was 94 +/- 41 kPa in the rigor state and 40 +/- 17 kPa in the relaxed state. The value of E( perpendicular) for calcium-activated myofibrils (55 +/- 29 kPa) was approximately a tenth that of Young's modulus in the longitudinal direction, a difference that at least partly reflects the transverse flexibility of the myosin molecule. PMID- 10692335 TI - Dynamic bending rigidity of a 200-bp DNA in 4 mM ionic strength: a transient polarization grating study. AB - DNA may exhibit three different kinds of bends: 1) permanent bends; 2) slowly relaxing bends due to fluctuations in a prevailing equilibrium between differently curved secondary conformations; and 3) rapidly relaxing dynamic bends within a single potential-of-mean-force basin. The dynamic bending rigidity (kappa(d)), or equivalently the dynamic persistence length, P(d) = kappa(d)/k(B)T, governs the rapidly relaxing bends, which are responsible for the flexural dynamics of DNA on a short time scale, t < or = 10(-5) s. However, all three kinds of bends contribute to the total equilibrium persistence length, P(tot), according to 1/P(tot) congruent with 1/P(pb) + 1/P(sr) + 1/P(d), where P(pb) is the contribution of the permanent bends and P(sr) is the contribution of the slowly relaxing bends. Both P(d) and P(tot) are determined for the same 200 bp DNA in 4 mM ionic strength by measuring its optical anisotropy, r(t), from 0 to 10 micros. Time-resolved fluorescence polarization anisotropy (FPA) measurements yield r(t) for DNA/ethidium complexes (1 dye/200 bp) from 0 to 120 ns. A new transient polarization grating (TPG) experiment provides r(t) for DNA/methylene blue complexes (1 dye/100 bp) over a much longer time span, from 20 ns to 10 micros. Accurate data in the very tail of the decay enable a model independent determination of the relaxation time (tau(R)) of the end-over-end tumbling motion, from which P(tot) = 500 A is estimated. The FPA data are used to obtain the best-fit pairs of P(d) and torsion elastic constant (alpha) values that fit those data equally well, and which are used to eliminate alpha as an independent variable. When the relevant theory is fitted to the entire TPG signal (S(t)), the end-over-end rotational diffusion coefficient is fixed at its measured value and alpha is eliminated in favor of P(d). Neither a true minimum in chi-squared nor a satisfactory fit could be obtained for P(d) anywhere in the range 500-5000 A, unless an adjustable amplitude of azimuthal wobble of the methylene blue was admitted. In that case, a well-defined global minimum and a reasonably good fit emerged at P(d) = 2000 A and (1/2) = 25 degrees. The discrimination against P(d) values <1600 A is very great. By combining the values, P(tot) = 500 A and P(d) = 2000 A with a literature estimate, P(pb) = 1370 A, a value P(sr) = 1300 A is estimated for the contribution of slowly relaxing bends. This value is analyzed in terms of a simple model in which the DNA is divided up into domains containing m bp, each of which experiences an all-or-none equilibrium between a straight and a uniformly curved conformation. With an appropriate estimate of the average bend angle per basepair of the curved conformation, a lower bound estimate, m = 55 bp, is obtained for the domain size of the coherently bent state. Previous measurements suggest that this coherent bend is not directional, or phase-locked, to the azimuthal orientation of the filament. PMID- 10692336 TI - Unraveling photoexcited conformational changes of bacteriorhodopsin by time resolved electron paramagnetic resonance spectroscopy. AB - By means of time-resolved electron paramagnetic resonance (EPR) spectroscopy, the photoexcited structural changes of site-directed spin-labeled bacteriorhodopsin are studied. A complete set of cysteine mutants of the C-D loop, positions 100 107, and of the E-F loop, including the first alpha-helical turns of helices E and F, positions 154-171, was modified with a methanethiosulfonate spin label. The EPR spectral changes occurring during the photocycle are consistent with a small movement of helix C and an outward tilt of helix F. These helix movements are accompanied by a rearrangement of the E-F loop and of the C-terminal turn of helix E. The kinetic analysis of the transient EPR data and the absorbance changes in the visible spectrum reveals that the conformational change occurs during the lifetime of the M intermediate. Prominent rearrangements of nitroxide side chains in the vicinity of D96 may indicate the preparation of the reprotonation of the Schiff base. All structural changes reverse with the recovery of the bacteriorhodopsin initial state. PMID- 10692337 TI - Structural changes of the sarcoplasmic reticulum Ca(2+)-ATPase upon nucleotide binding studied by fourier transform infrared spectroscopy. AB - Changes in the vibrational spectrum of the sarcoplasmic reticulum Ca(2+)-ATPase upon nucleotide binding were recorded in H(2)O and (2)H(2)O at -7 degrees C and pH 7.0. The reaction cycle was triggered by the photochemical release of nucleotides (ATP, ADP, and AMP-PNP) from a biologically inactive precursor (caged ATP, P(3)-1-(2-nitrophenyl) adenosine 5'-triphosphate, and related caged compounds). Infrared absorbance changes due to ATP release and two steps of the Ca(2+)-ATPase reaction cycle, ATP binding and phosphorylation, were followed in real time. Under the conditions used in our experiments, the rate of ATP binding was limited by the rate of ATP release (k(app) congruent with 3 s(-1) in H(2)O and k(app) congruent with 7 s(-1) in (2)H(2)O). Bands in the amide I and II regions of the infrared spectrum show that the conformation of the Ca(2+)-ATPase changes upon nucleotide binding. The observation of bands in the amide I region can be assigned to perturbations of alpha-helical and beta-sheet structures. According to similar band profiles in the nucleotide binding spectra, ATP, AMP PNP, and ADP induce similar conformational changes. However, subtle differences between ATP and AMP-PNP are observed; these are most likely due to the protonation state of the gamma-phosphate group. Differences between the ATP and ADP binding spectra indicate the significance of the gamma-phosphate group in the interactions between the Ca(2+)-ATPase and the nucleotide. Nucleotide binding affects Asp or Glu residues, and bands characteristic of their protonated side chains are observed at 1716 cm(-1) (H(2)O) and 1706 cm(-1) ((2)H(2)O) and seem to depend on the charge of the phosphate groups. Bands at 1516 cm(-1) (H(2)O) and 1514 cm(-1) ((2)H(2)O) are tentatively assigned to a protonated Tyr residue affected by nucleotide binding. Possible changes in Arg, Trp, and Lys absorption and in the nucleoside are discussed. The spectra are compared with those of nucleotide binding to arginine kinase, creatine kinase, and H-ras P21. PMID- 10692338 TI - Induced fit in arginine kinase. AB - Creatine kinase (CK) and arginine kinase (AK) are related enzymes that reversibly transfer a phosphoryl group between a guanidino compound and ADP. In the buffering of ATP energy levels, they are central to energy metabolism and have been paradigms of classical enzymology. Comparison of the open substrate-free structure of CK and the closed substrate-bound structure of AK reveals differences that are consistent with prior biophysical evidence of substrate induced conformational changes. Large and small domains undergo a hinged 13 degrees rotation. Several loops become ordered and adopt different positions in the presence of substrate, including one (residues 309-319) that moves 15 A to fold over the substrates. The conformational changes appear to be necessary in aligning the two substrates for catalysis, in configuring the active site only when productive phosphoryl transfer is possible, and excluding water from the active site to avoid wasteful ATP hydrolysis. PMID- 10692339 TI - Properties and crystal structure of a beta-barrel folding mutant. AB - A mutant of a beta-barrel protein, rat intestinal fatty acid binding protein, was predicted to be more stable than the wild-type protein due to a novel hydrogen bond. Equilibrium denaturation studies indicated the opposite: the V60N mutant protein was less stable. The folding transitions followed by CD and fluorescence were reversible and two-state for both mutant and wild-type protein. However, the rates of denaturation and renaturation of V60N were faster. During unfolding, the initial rate was associated with 75-80% of the fluorescence and all of the CD amplitude change. A subsequent rate accounted for the remaining fluorescence change for both proteins; thus the intermediate state lacked secondary structure. During folding, one rate was detected by both fluorescence and CD after an initial burst phase for both wild-type and mutant. An additional slower folding rate was detected by fluorescence for the mutant protein. The structure of the V60N mutant has been obtained and is nearly identical to prior crystal structures of IFABP. Analysis of mean differences in hydrogen bond and van der Waals interactions did not readily account for the stability loss due to the mutation. However, significant average differences of the solvent accessible surface and crystallographic displacement factors suggest entropic destabilization. PMID- 10692340 TI - Spectral fluctuation of a single fluorophore conjugated to a protein molecule. AB - We have measured the fluorescence spectra of a single fluorophore attached to a single protein molecule in aqueous solution using a total internal reflection fluorescence microscope. The most reactive cysteine residue of myosin subfragment 1 (S1) was labeled with tetramethylrhodamine. The spectral shift induced by a change in solvent from aqueous buffer to methanol in both single-molecule and bulk measurements were similar, indicating that, even at the single molecule level, the fluorescence spectrum is sensitive to microenvironmental changes of fluorophores. The time dependence of the fluorescence spectra of fluorophores attached to S1 molecules solely showed a fluctuation with time over a time scale of seconds. Because the fluorescence spectra of the same fluorophores directly conjugated to a glass surface remained constant, the spectral fluctuation observed for the fluorophores attached to S1 is most likely due to slow spontaneous conformational changes in the S1 molecule. Thus, single-molecule fluorescence spectroscopy appears to be a powerful tool to study the dynamic behavior of single biomolecules. PMID- 10692341 TI - Spectroscopy of individual light-harvesting 2 complexes of Rhodopseudomonas acidophila: diagonal disorder, intercomplex heterogeneity, spectral diffusion, and energy transfer in the B800 band. AB - This paper reports a detailed spectroscopic study of the B800 absorption band of individual light-harvesting 2 (LH2) complexes of the photosynthetic purple bacterium Rhodopseudomonas acidophila at 1. 2 K. By applying single-molecule detection techniques to this system, details and properties can be revealed that remain obscured in conventional ensemble experiments. For instance, from fluorescence-excitation spectra of the individual complexes a more direct measure of the diagonal disorder could be obtained. Further spectral diffusion phenomena and homogeneous linewidths of individual bacteriochlorophyll a (BChl a) molecules are observed, revealing valuable information on excited-state dynamics. This work demonstrates that it is possible to obtain detailed spectral information on individual pigment-protein complexes, providing direct insight into their electronic structure and into the mechanisms underlying the highly efficient energy transfer processes in these systems. PMID- 10692342 TI - Scanning electrochemical microscopy as a local probe of oxygen permeability in cartilage. AB - The use of scanning electrochemical microscopy, a high-resolution chemical imaging technique, to probe the distribution and mobility of solutes in articular cartilage is described. In this application, a mobile ultramicroelectrode is positioned close ( approximately 1 microm) to the cartilage sample surface, which has been equilibrated in a bathing solution containing the solute of interest. The solute is electrolyzed at a diffusion-limited rate, and the current response measured as the ultramicroelectrode is scanned across the sample surface. The topography of the samples was determined using Ru(CN)(6)(4-), a solute to which the cartilage matrix was impermeable. This revealed a number of pit-like depressions corresponding to the distribution of chondrocytes, which were also observed by atomic force and light microscopy. Subsequent imaging of the same area of the cartilage sample for the diffusion-limited reduction of oxygen indicated enhanced, but heterogeneous, permeability of oxygen across the cartilage surface. In particular, areas of high permeability were observed in the cellular and pericellular regions. This is the first time that inhomogeneities in the permeability of cartilage toward simple solutes, such as oxygen, have been observed on a micrometer scale. PMID- 10692343 TI - One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins. AB - We have used one- (OPE) and two-photon (TPE) excitation with time-correlated single-photon counting techniques to determine time-resolved fluorescence intensity and anisotropy decays of the wild-type Green Fluorescent Protein (GFP) and two red-shifted mutants, S65T-GFP and RSGFP. WT-GFP and S65T-GFP exhibited a predominant approximately 3 ns monoexponential fluorescence decay, whereas for RSGFP the main lifetimes were approximately 1.1 ns (main component) and approximately 3.3 ns. The anisotropy decay of WT-GFP and S65T-GFP was also monoexponential (global rotational correlation time of 16 +/- 1 ns). The approximately 1.1 ns lifetime of RSGFP was associated with a faster rotational depolarization, evaluated as an additional approximately 13 ns component. This feature we attribute tentatively to a greater rotational freedom of the anionic chromophore. With OPE, the initial anisotropy was close to the theoretical limit of 0.4; with TPE it was higher, approaching the TPE theoretical limit of 0.57 for the colinear case. The measured power dependence of the fluorescence signals provided direct evidence for TPE. The general independence of fluorescence decay times, rotation correlation times, and steady-state emission spectra on the excitation mode indicates that the fluorescence originated from the same distinct excited singlet states (A*, I*, B*). However, we observed a relative enhancement of blue fluorescence peaked at approximately 440 nm for TPE compared to OPE, indicating different relative excitation efficiencies. We infer that the two lifetimes of RSGFP represent the deactivation of two substates of the deprotonated intermediate (I*), distinguished by their origin (i.e., from A* or B*) and by nonradiative decay rates reflecting different internal environments of the excited-state chromophore. PMID- 10692344 TI - High-resolution imaging of antibodies by tapping-mode atomic force microscopy: attractive and repulsive tip-sample interaction regimes. AB - A force microscope operated with an amplitude modulation feedback (usually known as tapping-mode atomic force microscope) has two tip-sample interaction regimes, attractive and repulsive. We have studied the performance of those regimes to imaging single antibody molecules. The attractive interaction regime allows determination of the basic morphologies of the antibodies on the support. More importantly, this regime is able to resolve the characteristic Y-shaped domain structure of antibodies and the hinge region between domains. Imaging in the repulsive interaction regime is associated with the irreversible deformation of the molecules. This causes a significant loss in resolution and contrast. Two major physical differences distinguish the repulsive interaction regime from the attractive interaction regime: the existence of tip-sample contact and the strength of the forces involved. PMID- 10692345 TI - Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling. AB - A new method for the size-distribution analysis of polymers by sedimentation velocity analytical ultracentrifugation is described. It exploits the ability of Lamm equation modeling to discriminate between the spreading of the sedimentation boundary arising from sample heterogeneity and from diffusion. Finite element solutions of the Lamm equation for a large number of discrete noninteracting species are combined with maximum entropy regularization to represent a continuous size-distribution. As in the program CONTIN, the parameter governing the regularization constraint is adjusted by variance analysis to a predefined confidence level. Estimates of the partial specific volume and the frictional ratio of the macromolecules are used to calculate the diffusion coefficients, resulting in relatively high-resolution sedimentation coefficient distributions c(s) or molar mass distributions c(M). It can be applied to interference optical data that exhibit systematic noise components, and it does not require solution or solvent plateaus to be established. More details on the size-distribution can be obtained than from van Holde-Weischet analysis. The sensitivity to the values of the regularization parameter and to the shape parameters is explored with the help of simulated sedimentation data of discrete and continuous model size distributions, and by applications to experimental data of continuous and discrete protein mixtures. PMID- 10692346 TI - Factors governing the assembly of cationic phospholipid-DNA complexes. AB - The interaction of DNA with a novel cationic phospholipid transfection reagent, 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EDOPC), was investigated by monitoring thermal effects, particle size, vesicle rupture, and lipid mixing. By isothermal titration calorimetry, the heat of interaction between large unilamellar EDOPC vesicles and plasmid DNA was endothermic at both physiological and low ionic strength, although the heat absorbed was slightly larger at the higher ionic strength. The energetic driving force for DNA-EDOPC association is thus an increase in entropy, presumably due to release of counterions and water. The estimated minimum entropy gain per released counterion was 1.4 cal/mole- degrees K (about 0.7 kT), consistent with previous theoretical predictions. All experimental approaches revealed significant differences in the DNA-lipid particle, depending upon whether complexes were formed by the addition of DNA to lipid or vice versa. When EDOPC vesicles were titrated with DNA at physiological ionic strength, particle size increased, vesicles ruptured, and membrane lipids became mixed as the amount of DNA was added up to a 1.6:1 (+:-) charge ratio. This charge ratio also corresponded to the calorimetric end point. In contrast, when lipid was added to DNA, vesicles remained separate and intact until a charge ratio of 1:1 (+:-) was exceeded. Under such conditions, the calorimetric end point was 3:1 (+:-). Thus it is clear that fundamental differences in DNA cationic lipid complexes exist, depending upon their mode of formation. A model is proposed to explain the major differences between these two situations. Significant effects of ionic strength were observed; these are rationalized in terms of the model. The implications of the analysis are that considerable control can be exerted over the structure of the complex by exploiting vectorial preparation methods and manipulating ionic strength. PMID- 10692347 TI - Detergent-induced conformational changes of Humicola lanuginosa lipase studied by fluorescence spectroscopy. AB - Detergent (pentaoxyethylene octyl ether, C(8)E(5))-induced conformational changes of Humicola lanuginosa lipase (HLL) were investigated by stationary and time resolved fluorescence intensity and anisotropy measurements. Activation of HLL is characterized by opening of a surface loop (the "lid") residing directly over the enzyme active site. The interaction of HLL with C(8)E(5) increases fluorescence intensities, prolongs fluorescence lifetimes, and decreases the values of steady state anisotropy, residual anisotropy, and the short rotational correlation time. Based on these data, we propose the following model. Already below critical micellar concentration (CMC) the detergent can intercalate into the active site accommodating cleft, while the lid remains closed. Occupation of the cleft by C(8)E(5) also blocks the entry of the monomeric substrate, and inhibition of catalytic activity at [C(8)E(5)] less than or equal to CMC is evident. At a threshold concentration close to CMC the cooperativity of the hydrophobicity driven binding of C(8)E(5) to the lipase increases because of an increase in the number of C(8)E(5) molecules present in the premicellar nucleates on the hydrophobic surface of HLL. These aggregates contacting the lipase should have long enough residence times to allow the lid to open completely and expose the hydrophobic cleft. Concomitantly, the cleft becomes filled with C(8)E(5) and the "open" conformation of HLL becomes stable. PMID- 10692348 TI - Growing an actin gel on spherical surfaces. AB - Inspired by the motility of the bacteria Listeria monocytogenes, we have experimentally studied the growth of an actin gel around spherical beads grafted with ActA, a protein known to be the promoter of bacteria movement. On ActA grafted beads F-actin is formed in a spherical manner, whereas on the bacteria a "comet-like" tail of F-actin is produced. We show experimentally that the stationary thickness of the gel depends on the radius of the beads. Moreover, the actin gel is not formed if the ActA surface density is too low. To interpret our results, we propose a theoretical model to explain how the mechanical stress (due to spherical geometry) limits the growth of the actin gel. Our model also takes into account treadmilling of actin. We deduce from our work that the force exerted by the actin gel on the bacteria is of the order of 10 pN. Finally, we estimate from our theoretical model possible conditions for developing actin comet tails. PMID- 10692349 TI - Signaling components in bacterial locomotion and sensory reception. PMID- 10692350 TI - Three new NifA-regulated genes in the Bradyrhizobium japonicum symbiotic gene region discovered by competitive DNA-RNA hybridization. AB - The so-called symbiotic region of the Bradyrhizobium japonicum chromosome (C. Kundig, H. Hennecke, and M. Gottfert, J. Bacteriol. 175:613-622, 1993) was screened for the presence of genes controlled by the nitrogen fixation regulatory protein NifA. Southern blots of restriction enzyme-digested cosmids that represent an ordered, overlapping library of the symbiotic region were competitively hybridized with in vitro-labeled RNA from anaerobically grown wild type cells and an excess of RNA isolated either from anaerobically grown nifA and rpoN mutant cells or from aerobically grown wild-type cells. In addition to the previously characterized nif and fix gene clusters, we identified three new NifA regulated genes that were named nrgA, nrgB, and nrgC (nrg stands for NifA regulated gene). The latter two probably form an operon, nrgBC. The proteins encoded by nrgC and nrgA exhibited amino acid sequence similarity to bacterial hydroxylases and N-acetyltransferases, respectively. The product of nrgB showed no significant similarity to any protein with a database entry. Primer extension experiments and expression studies with translational lacZ fusions revealed the presence of a functional -24/-12-type promoter upstream of nrgA and nrgBC and proved the NifA- and RpoN (sigma(54))-dependent transcription of the respective genes. Null mutations introduced into nrgA and nrgBC resulted in mutant strains that exhibited wild-type-like symbiotic properties, including nitrogen fixation, when tested on soybean, cowpea, or mung bean host plants. Thus, the discovery of nrgA and nrgBC further emphasizes the previously suggested role of NifA as an activator of anaerobically induced genes other than the classical nitrogen fixation genes. PMID- 10692351 TI - Functional analysis of PvdS, an iron starvation sigma factor of Pseudomonas aeruginosa. AB - In Pseudomonas aeruginosa, iron modulates gene expression through a cascade of negative and positive regulatory proteins. The master regulator Fur is involved in iron-dependent repression of several genes. One of these genes, pvdS, was predicted to encode a putative sigma factor responsible for the transcription of a subset of genes of the Fur regulon. PvdS appears to belong to a structurally and functionally distinct subgroup of the extracytoplasmic function family of alternative sigma factors. Members of this subgroup, also including PbrA from Pseudomonas fluorescens, PfrI and PupI from Pseudomonas putida, and FecI from Escherichia coli, are controlled by the Fur repressor, and they activate transcription of genes for the biosynthesis or the uptake of siderophores. Evidence is provided that the PvdS protein of P. aeruginosa is endowed with biochemical properties of eubacterial sigma factors, as it spontaneously forms 1:1 complexes with the core fraction of RNA polymerase (RNAP, alpha(2)betabeta' subunits), thereby promoting in vitro binding of the PvdS-RNAP holoenzyme to the promoter region of the pvdA gene. These functional features of PvdS are consistent with the presence of structural domains predicted to be involved in core RNAP binding, promoter recognition, and open complex formation. The activity of pyoverdin biosynthetic (pvd) promoters was significantly lower in E. coli overexpressing the multicopy pvdS gene than in wild-type P. aeruginosa PAO1 carrying the single gene copy, and pvd::lacZ transcriptional fusions were silent in both pfrI (the pvdS homologue) and pfrA (a positive regulator of pseudobactin biosynthetic genes) mutants of P. putida WCS358, while they are expressed at PAO1 levels in wild-type WCS358. Moreover, the PvdS-RNAP holoenzyme purified from E. coli lacked the ability to generate in vitro transcripts from the pvdA promoter. These observations suggest that at least one additional positive regulator could be required for full activity of the PvdS-dependent transcription complex both in vivo and in vitro. This is consistent with the presence of a putative activator binding site (the iron starvation box) at variable distance from the transcription initiation sites of promoters controlled by the iron starvation sigma factors PvdS, PfrI, and PbrA of fluorescent pseudomonads. PMID- 10692352 TI - The TetA(K) tetracycline/H(+) antiporter from Staphylococcus aureus: mutagenesis and functional analysis of motif C. AB - Conserved motif C, identified within members of the major facilitator superfamily (MFS) of transport proteins that mediate drug export, was examined in the tetracycline resistance efflux protein TetA(K) from Staphylococcus aureus; motif C is contained within transmembrane segment 5. Using site-directed mutagenesis, the importance of the conserved glycine (G151, G155, G159, and G160) and proline (P156) residues within this motif was investigated. Over 40 individual amino acid replacements were introduced; however, only alanine and serine substitutions for glycine at G151, G155, and G160 were found to retain significant levels of tetracycline resistance and transport activity in cells expressing mutant proteins. Notably, P156 and G159 appear to be crucial, as amino acid replacements at these positions either significantly reduced or abolished tetracycline/H(+) activity. The highly conserved nature of motif C and its distribution throughout drug exporters imply that the residues of motif C play a similar role in all MFS proteins that function as antiporters. PMID- 10692353 TI - A novel spore peptidoglycan hydrolase of Bacillus cereus: biochemical characterization and nucleotide sequence of the corresponding gene, sleL. AB - The exudate of germinated spores of B. cereus IFO 13597 in 0.15 M KCl-50 mM potassium phosphate (pH 7.0) contained a spore-lytic enzyme which has substrate specificity for fragmented spore cortex from wild-type organisms (cortical fragment-lytic enzyme [CFLE]), in addition to a previously characterized germination-specific hydrolase which acts on intact spore cortex (spore cortex lytic enzyme [SCLE]) (R. Moriyama, S. Kudoh, S. Miyata, S. Nonobe, A. Hattori, and S. Makino, J. Bacteriol. 178:5330-5332, 1996). CFLE was not capable of degrading isolated cortical fragments from spores of Bacillus subtilis ADD1, which lacks muramic acid delta-lactam. This suggests that CFLE cooperates with SCLE in cortex hydrolysis during germination. CFLE was purified in an active form and identified as a 48-kDa protein which functions as an N-acetylglucosaminidase. Immunochemical studies suggested that the mature enzyme is localized on a rather peripheral region of the dormant spore, probably the exterior of the cortex layer. A gene encoding the enzyme, sleL, was cloned in Escherichia coli, and the nucleotide sequence was determined. The gene encodes a protein of 430 amino acids with a deduced molecular weight of 48,136. The N-terminal region contains a repeated motif common to several peptidoglycan binding proteins. Inspection of the data banks showed no similarity of CFLE with N-acetylglucosaminidases found so far, suggesting that CFLE is a novel type of N-acetylglucosaminidase. The B. subtilis genome sequence contains genes, yaaH and ydhD, which encode putative proteins showing similarity to SleL. PMID- 10692354 TI - A gene cluster involved in metal homeostasis in the cyanobacterium Synechocystis sp. strain PCC 6803. AB - A gene cluster composed of nine open reading frames (ORFs) involved in Ni(2+), Co(2+), and Zn(2+) sensing and tolerance in the cyanobacterium Synechocystis sp. strain PCC 6803 has been identified. The cluster includes an Ni(2+) response operon and a Co(2+) response system, as well as a Zn(2+) response system previously described. Expression of the Ni(2+) response operon (nrs) was induced in the presence of Ni(2+) and Co(2+). Reduced Ni(2+) tolerance was observed following disruption of two ORFs of the operon (nrsA and nrsD). We also show that the nrsD gene encodes a putative Ni(2+) permease whose carboxy-terminal region is a metal binding domain. The Co(2+) response system is composed of two divergently transcribed genes, corR and corT, mutants of which showed decreased Co(2+) tolerance. Additionally, corR mutants showed an absence of Co(2+)-dependent induction of corT, indicating that CorR is a transcriptional activator of corT. To our knowledge, CorR is the first Co(2+)-sensing transcription factor described. Our data suggest that this region of the Synechocystis sp. strain PCC 6803 genome is involved in sensing and homeostasis of Ni(2+), Co(2+), and Zn(2+). PMID- 10692355 TI - Evolution of drug resistance in experimental populations of Candida albicans. AB - Adaptation to inhibitory concentrations of the antifungal agent fluconazole was monitored in replicated experimental populations founded from a single, drug sensitive cell of the yeast Candida albicans and reared over 330 generations. The concentration of fluconazole was maintained at twice the MIC in six populations; no fluconazole was added to another six populations. All six replicate populations grown with fluconazole adapted to the presence of drug as indicated by an increase in MIC; none of the six populations grown without fluconazole showed any change in MIC. In all populations evolved with drug, increased fluconazole resistance was accompanied by increased resistance to ketoconazole and itraconazole; these populations contained ergosterol in their cell membranes and were amphotericin sensitive. The increase in fluconazole MIC in the six populations evolved with drug followed different trajectories, and these populations achieved different levels of resistance, with distinct overexpression patterns of four genes involved in azole resistance: the ATP-binding cassette transporter genes, CDR1 and CDR2; the gene encoding the target enzyme of the azoles in the ergosterol biosynthetic pathway, ERG11; and the major facilitator gene, MDR1. Selective sweeps in these populations were accompanied by additional genomic changes with no known relationship to drug resistance: loss of heterozygosity in two of the five marker genes assayed and alterations in DNA fingerprints and electrophoretic karyotypes. These results show that chance, in the form of mutations that confer an adaptive advantage, is a determinant in the evolution of azole drug resistance in experimental populations of C. albicans. PMID- 10692356 TI - Molecular basis for the temperature sensitivity of Escherichia coli pth(Ts). AB - The gene pth, encoding peptidyl-tRNA hydrolase (Pth), is essential for protein synthesis and viability of Escherichia coli. Two pth mutants have been studied in depth: a pth(Ts) mutant isolated as temperature sensitive and a pth(rap) mutant selected as nonpermissive for bacteriophage lambda vegetative growth. Here we show that each mutant protein is defective in a different way. The Pth(Ts) protein was very unstable in vivo, both at 43 degrees C and at permissive temperatures, but its specific activity was comparable to that of the wild-type enzyme, Pth(wt). Conversely, the mutant Pth(rap) protein had the same stability as Pth(wt), but its specific activity was low. The thermosensitivity of the pth(Ts) mutant, presumably, ensues after Pth(Ts) protein levels are reduced at 43 degrees C. Conditions that increased the cellular Pth(Ts) concentration, a rise in gene copy number or diminished protein degradation, allowed cell growth at a nonpermissive temperature. Antibiotic-mediated inhibition of mRNA and protein synthesis, but not of peptidyl-tRNA drop-off, reduced pth(Ts) cell viability even at a permissive temperature. Based on these results, we suggest that Pth(Ts) protein, being unstable in vivo, supports cell viability only if its concentration is maintained above a threshold that allows general protein synthesis. PMID- 10692357 TI - The RofA binding site in Streptococcus pyogenes is utilized in multiple transcriptional pathways. AB - Understanding the regulation of adhesins defines a pathogenic bacterium's interaction with the local environment within the host. In certain strains of Streptococcus pyogenes, transcription of prtF, the gene which encodes the fibronectin-binding adhesin protein F, is activated by RofA under anaerobic conditions. RofA binds specifically to DNA in its target promoters and autoregulates its own expression. In this study, we have used DNase I protection assays to further investigate the interaction of RofA with its target promoters. In the region between rofA and the gene which encodes protein F (prtF), RofA binds to two distinct sites: a smaller site (17 bp) adjacent to the rofA promoter, and a larger site (40 bp) adjacent to the prtF promoter. Analysis of fusions to a novel reporter gene whose product consists of the fusion of the N terminal secretion domain of protein F with the C-terminal enzymatic domain of the enterococcal alkaline phosphatase (PhoZ) revealed that the small RofA binding site had no direct role in control of prtF transcription but contributed to regulation of rofA. Comparison in several strains representing different patterns of prtF expression indicated that the larger site was required for activation of rofA and of prtF in all strains by both RofA-dependent and -independent pathways. Thus, it would appear that a common recognition sequence provides separate entries to a final common pathway in S. pyogenes virulence gene expression. The identification of multiple RofA-like proteins and promoters with RofA binding sites implies the existence of a widespread interacting regulatory network. PMID- 10692358 TI - TraG from RP4 and TraG and VirD4 from Ti plasmids confer relaxosome specificity to the conjugal transfer system of pTiC58. AB - Plasmid conjugation systems are composed of two components, the DNA transfer and replication system, or Dtr, and the mating pair formation system, or Mpf. During conjugal transfer an essential factor, called the coupling protein, is thought to interface the Dtr, in the form of the relaxosome, with the Mpf, in the form of the mating bridge. These proteins, such as TraG from the IncP1 plasmid RP4 (TraG(RP4)) and TraG and VirD4 from the conjugal transfer and T-DNA transfer systems of Ti plasmids, are believed to dictate specificity of the interactions that can occur between different Dtr and Mpf components. The Ti plasmids of Agrobacterium tumefaciens do not mobilize vectors containing the oriT of RP4, but these IncP1 plasmid derivatives lack the trans-acting Dtr functions and TraG(RP4). A. tumefaciens donors transferred a chimeric plasmid that contains the oriT and Dtr genes of RP4 and the Mpf genes of pTiC58, indicating that the Ti plasmid mating bridge can interact with the RP4 relaxosome. However, the Ti plasmid did not mobilize transfer from an IncQ relaxosome. The Ti plasmid did mobilize such plasmids if TraG(RP4) was expressed in the donors. Mutations in traG(RP4) with defined effects on the RP4 transfer system exhibited similar phenotypes for Ti plasmid-mediated mobilization of the IncQ vector. When provided with VirD4, the tra system of pTiC58 mobilized plasmids from the IncQ relaxosome. However, neither TraG(RP4) nor VirD4 restored transfer to a traG mutant of the Ti plasmid. VirD4 also failed to complement a traG(RP4) mutant for transfer from the RP4 relaxosome or for RP4-mediated mobilization from the IncQ relaxosome. TraG(RP4)-mediated mobilization of the IncQ plasmid by pTiC58 did not inhibit Ti plasmid transfer, suggesting that the relaxosomes of the two plasmids do not compete for the same mating bridge. We conclude that TraG(RP4) and VirD4 couples the IncQ but not the Ti plasmid relaxosome to the Ti plasmid mating bridge. However, VirD4 cannot couple the IncP1 or the IncQ relaxosome to the RP4 mating bridge. These results support a model in which the coupling proteins specify the interactions between Dtr and Mpf components of mating systems. PMID- 10692359 TI - Association of the cytoplasmic membrane protein XpsN with the outer membrane protein XpsD in the type II protein secretion apparatus of Xanthomonas campestris pv. campestris. AB - An xps gene cluster composed of 11 open reading frames is required for the type II protein secretion in Xanthomonas campestris pv. campestris. Immediately upstream of the xpsD gene, which encodes an outer membrane protein that serves as the secretion channel by forming multimers, there exists an open reading frame (previously designated ORF2) that could encode a protein of 261 amino acid residues. Its N-terminal hydrophobic region is a likely membrane-anchoring sequence. Antibody raised against this protein could detect in the wild-type strain of X. campestris pv. campestris a protein band with an apparent molecular mass of 36 kDa by Western blotting. Its aberrant slow migration in sodium dodecyl sulfate-polyacrylamide gels might be due to its high proline content. We designated this protein XpsN. By constructing a mutant strain with an in-frame deletion of the chromosomal xpsN gene, we demonstrated that it is required for the secretion of extracellular enzyme by X. campestris pv. campestris. Subcellular fractionation studies indicated that the XpsN protein was tightly associated with the membrane. Sucrose gradient sedimentation followed by immunoblot analysis revealed that it primarily appeared in the cytoplasmic membrane fractions. Immune precipitation experiments indicated that the XpsN protein was coprecipitated with the XpsD protein. In addition, the XpsN protein was co-eluted with the (His)(6)-tagged XpsD protein from the metal affinity chromatography column. All observations suggested that the XpsN protein forms a stable complex with the XpsD protein. In addition, immune precipitation analysis of the XpsN protein with various truncated XpsD proteins revealed that the C terminal region of the XpsD protein between residues 650 and 759 was likely to be involved in complex formation between the two. PMID- 10692360 TI - ssrA (tmRNA) plays a role in Salmonella enterica serovar Typhimurium pathogenesis. AB - Escherichia coli ssrA encodes a small stable RNA molecule, tmRNA, that has many diverse functions, including tagging abnormal proteins for degradation, supporting phage growth, and modulating the activity of DNA binding proteins. Here we show that ssrA plays a role in Salmonella enterica serovar Typhimurium pathogenesis and in the expression of several genes known to be induced during infection. Moreover, the phage-like attachment site, attL, encoded within ssrA, serves as the site of integration of a region of Salmonella-specific sequence; adjacent to the 5' end of ssrA is another region of Salmonella-specific sequence with extensive homology to predicted proteins encoded within the unlinked Salmonella pathogenicity island SPI4. S. enterica serovar Typhimurium ssrA mutants fail to support the growth of phage P22 and are delayed in their ability to form viable phage particles following induction of a phage P22 lysogen. These data indicate that ssrA plays a role in the pathogenesis of Salmonella, serves as an attachment site for Salmonella-specific sequences, and is required for the growth of phage P22. PMID- 10692361 TI - Components of the RP4 conjugative transfer apparatus form an envelope structure bridging inner and outer membranes of donor cells: implications for related macromolecule transport systems. AB - During bacterial conjugation, the single-stranded DNA molecule is transferred through the cell envelopes of the donor and the recipient cell. A membrane spanning transfer apparatus encoded by conjugative plasmids has been proposed to facilitate protein and DNA transport. For the IncPalpha plasmid RP4, a thorough sequence analysis of the gene products of the transfer regions Tra1 and Tra2 revealed typical features of mainly inner membrane proteins. We localized essential RP4 transfer functions to Escherichia coli cell fractions by immunological detection with specific polyclonal antisera. Each of the gene products of the RP4 mating pair formation (Mpf) system, specified by the Tra2 core region and by traF of the Tra1 region, was found in the outer membrane fraction with one exception, the TrbB protein, which behaved like a soluble protein. The membrane preparation from Mpf-containing cells had an additional membrane fraction whose density was intermediate between those of the cytoplasmic and outer membranes, suggesting the presence of attachment zones between the two E. coli membranes. The Tra1 region is known to encode the components of the RP4 relaxosome. Several gene products of this transfer region, including the relaxase TraI, were detected in the soluble fraction, but also in the inner membrane fraction. This indicates that the nucleoprotein complex is associated with and/or assembled facing the cytoplasmic site of the E. coli cell envelope. The Tra1 protein TraG was predominantly localized to the cytoplasmic membrane, supporting its potential role as an interface between the RP4 Mpf system and the relaxosome. PMID- 10692362 TI - Identification of the active site of HetR protease and its requirement for heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120. AB - HetR is a serine-type protease required for heterocyst differentiation in heterocystous cyanobacteria under conditions of nitrogen deprivation. We have identified the active Ser residue of HetR from Anabaena sp. strain PCC 7120 by site-specific mutagenesis. By changing the S152 residue to an Ala residue, the mutant protein cannot be labeled by Dansyl fluoride, a specific serine-type protein inhibitor. The mutant protein showed no autodegradation in vitro. The mutant hetR gene was introduced into Anabaena strain 884a, a hetR mutant. The resultant strain, Anabaena strain S152A, could not form heterocysts under conditions of nitrogen deprivation even though the up-regulation of the mutant hetR gene was induced upon removal of combined nitrogen. The Anabaena strain 216, which carries a mutant hetR gene encoding S179N HetR and could not form heterocysts, also produced HetR protein upon induction. Sequence comparison shows that Ser152 is conserved in all cyanobacterial HetR. Immunoblotting was used to study HetR induction in both the wild-type and mutant strains. The amount of mutant HetR in strain S152A and in strain 216 increased continuously for 24 h after nitrogen step-down, while the amount of HetR in wild-type cells reached a maximum level within 6 h after nitrogen step-down. Our results show the Ser152 is the active site of HetR. The protease activity is required for heterocyst differentiation and might be needed for repression of HetR overproduction under conditions of nitrogen deprivation. PMID- 10692363 TI - EFG1 null mutants of Candida albicans switch but cannot express the complete phenotype of white-phase budding cells. AB - The Candida albicans gene EFG1 encodes a putative trans-acting factor. In strain WO-1, which undergoes the white-opaque transition, EFG1 is transcribed as a 3.2 kb mRNA in white-phase cells and a less-abundant 2.2-kb mRNA in opaque-phase cells. cDNA sequencing and 5' rapid amplification of cDNA ends analysis demonstrate that the major difference in molecular mass of the two transcripts is due to different transcription start sites. EFG1 null mutants form opaque-phase colonies and express the opaque-phase cell phenotype at 25 degrees C. When shifted from 25 to 42 degrees C, mutant opaque-phase cells undergo phenotypic commitment to the white phase, which includes deactivation of the opaque-phase specific gene OP4 and activation of the white-phase-specific gene WH11, as do wild-type opaque-phase cells. After the commitment event, EFG1 null mutant cells form daughter cells which have the smooth (pimpleless) surface of white-phase cells but the elongate morphology of opaque-phase cells. Taken together, these results demonstrate that EFG1 expression is not essential for the switch event per se, but is essential for a subset of phenotypic characteristics necessary for the full expression of the phenotype of white-phase cells. These results demonstrate that EFG1 is not the site of the switch event, but is, rather, downstream of the switch event. PMID- 10692364 TI - Expression of ykdA, encoding a Bacillus subtilis homologue of HtrA, is heat shock inducible and negatively autoregulated. AB - There are three members of the HtrA family of serine proteases, YkdA, YvtA, and YyxA, encoded in the chromosome of Bacillus subtilis. In this study, we report on the promoter structure and regulation of ykdA expression. The ykdA gene is heat inducible, exhibiting a biphasic pattern of expression during a 60-min interval after heat shock. Increased expression after heat shock occurs at the transcriptional level. The heat-shock-inducible promoter has a single mismatch with a SigA-type -10 motif, but does not exhibit similarity to a SigA -35 region. There are six octamer repeats with a consensus TTTTCACA positioned at, and upstream of, the normal position of a -35 region. While repeats V and VI appear dispensable, repeat IV is essential for normal thermoinducible expression. This promoter structure is also found in the control region of yvtA, encoding a second member of this family of proteases. Expression of ykdA is negatively autoregulated both during the growth cycle and during heat shock. Our evidence suggests that YkdA protease activity is not required for this form of regulation. Null mutants of ykdA display increased tolerance to heat and are 80-fold more resistant to 10 mM hydrogen peroxide than wild-type cells. However, ykdA expression is not induced by hydrogen peroxide. These results indicate that the regulon to which YkdA belongs is linked to the oxidative stress response in B. subtilis. PMID- 10692365 TI - Specificity mutants of the binding protein of the oligopeptide transport system of Lactococcus lactis. AB - The kinetic properties of wild-type and mutant oligopeptide binding proteins of Lactococcus lactis were determined. To observe the properties of the mutant proteins in vivo, the oppA gene was deleted from the chromosome of L. lactis to produce a strain that was totally defective in oligopeptide transport. Amplified expression of the oppA gene resulted in an 8- to 12-fold increase in OppA protein relative to the wild-type level. The amplified expression was paralleled by increased bradykinin binding activity, but had relatively little effect on the overall transport of bradykinin via Opp. Several site-directed mutants were constructed on the basis of a comparison of the primary sequences of OppA from Salmonella enterica serovar Typhimurium and L. lactis, taking into account the known structure of the serovar Typhimurium protein. Putative peptide binding-site residues were mutated. All the mutant OppA proteins exhibited a decreased binding affinity for the high-affinity peptide bradykinin. Except for OppA(D471R), the mutant OppA proteins displayed highly defective bradykinin uptake, whereas the transport of the low-affinity substrate KYGK was barely affected. Cells expressing OppA(D471R) had a similar K(m) for transport, whereas the V(max) was increased more than twofold as compared to the wild-type protein. The data are discussed in the light of a kinetic model and imply that the rate of transport is determined to a large extent by the donation of the peptide from the OppA protein to the translocator complex. PMID- 10692366 TI - Evolution of arginine biosynthesis in the bacterial domain: novel gene-enzyme relationships from psychrophilic Moritella strains (Vibrionaceae) and evolutionary significance of N-alpha-acetyl ornithinase. AB - In the arginine biosynthetic pathway of the vast majority of prokaryotes, the formation of ornithine is catalyzed by an enzyme transferring the acetyl group of N-alpha-acetylornithine to glutamate (ornithine acetyltransferase [OATase]) (argJ encoded). Only two exceptions had been reported-the Enterobacteriaceae and Myxococcus xanthus (members of the gamma and delta groups of the class Proteobacteria, respectively)-in which ornithine is produced from N-alpha acetylornithine by a deacylase, acetylornithinase (AOase) (argE encoded). We have investigated the gene-enzyme relationship in the arginine regulons of two psychrophilic Moritella strains belonging to the Vibrionaceae, a family phylogenetically related to the Enterobacteriaceae. Most of the arg genes were found to be clustered in one continuous sequence divergently transcribed in two wings, argE and argCBFGH(A) ["H(A)" indicates that the argininosuccinase gene consists of a part homologous to known argH sequences and of a 3' extension able to complement an Escherichia coli mutant deficient in the argA gene, encoding N alpha-acetylglutamate synthetase, the first enzyme committed to the pathway]. Phylogenetic evidence suggests that this new clustering pattern arose in an ancestor common to Vibrionaceae and Enterobacteriaceae, where OATase was lost and replaced by a deacylase. The AOase and ornithine carbamoyltransferase of these psychrophilic strains both display distinctly cold-adapted activity profiles, providing the first cold-active examples of such enzymes. PMID- 10692367 TI - Functionality of purified sigma(N) (sigma(54)) and a NifA-like protein from the hyperthermophile Aquifex aeolicus. AB - The genome sequence of the extremely thermophilic bacterium Aquifex aeolicus encodes alternative sigma factor sigma(N) (sigma(54), RpoN) and five potential sigma(N)-dependent transcriptional activators. Although A. aeolicus possesses no recognizable nitrogenase genes, two of the activators have a high degree of sequence similarity to NifA proteins from nitrogen-fixing proteobacteria. We identified five putative sigma(N)-dependent promoters upstream of operons implicated in functions including sulfur respiration, nitrogen assimilation, nitrate reductase, and nitrite reductase activity. We cloned, overexpressed (in Escherichia coli), and purified A. aeolicus sigma(N) and the NifA homologue, AQ_218. Purified A. aeolicus sigma(N) bound to E. coli core RNA polymerase and bound specifically to a DNA fragment containing E. coli promoter glnHp2 and to several A. aeolicus DNA fragments containing putative sigma(N)-dependent promoters. When combined with E. coli core RNA polymerase, A. aeolicus sigma(N) supported A. aeolicus NifA-dependent transcription from the glnHp2 promoter. The E. coli activator PspFDeltaHTH did not stimulate transcription. The NifA homologue, AQ_218, bound specifically to a DNA sequence centered about 100 bp upstream of the A. aeolicus glnBA operon and so is likely to be involved in the regulation of nitrogen assimilation in this organism. These results argue that the sigma(N) enhancer-dependent transcription system operates in at least one extreme environment, and that the activator and sigma(N) have coevolved. PMID- 10692368 TI - Transcriptional and mutational analysis of the uptake hydrogenase of the filamentous cyanobacterium Anabaena variabilis ATCC 29413. AB - A 10-kb DNA region of the cyanobacterium Anabaena variabilis ATCC 29413 containing the structural genes of the uptake hydrogenase (hupSL) was cloned and sequenced. In contrast to the hupL gene of Anabaena sp. strain PCC 7120, which is interrupted by a 10.5-kb DNA fragment in vegetative cells, there is no programmed rearrangement within the hupL gene during the heterocyst differentiation of A. variabilis. The hupSL genes were transcribed as a 2.7-kb operon and were induced only under nitrogen-fixing conditions, as shown by Northern blot experiments and reverse transcriptase PCR. Primer extension experiments with a fluorescence labeled oligonucleotide primer confirmed these results and identified the 5' start of the mRNA transcript 103 bp upstream of the ATG initiation codon. A consensus sequence in the promoter that is recognized by the fumarate nitrate reductase regulator (Fnr) could be detected. The hupSL operon in A. variabilis was interrupted by an interposon deletion (mutant strain AVM13). Under N(2) fixing conditions, the mutant strain exhibited significantly increased rates in H(2) accumulation and produced three times more hydrogen than the wild type. These results indicate that the uptake hydrogenase is catalytically active in the wild type and that the enzyme reoxidizes the H(2) developed by the nitrogenase. The Nif phenotype of the mutant strain showed a slight decrease of acetylene reduction compared to that of the wild type. PMID- 10692369 TI - Global regulatory mutations in csrA and rpoS cause severe central carbon stress in Escherichia coli in the presence of acetate. AB - The csrA gene encodes a small RNA-binding protein, which acts as a global regulator in Escherichia coli and other bacteria (T. Romeo, Mol. Microbiol. 29:1321-1330, 1998). Its key regulatory role in central carbon metabolism, both as an activator of glycolysis and as a potent repressor of glycogen biosynthesis and gluconeogenesis, prompted us to examine the involvement of csrA in acetate metabolism and the tricarboxylic acid (TCA) cycle. We found that growth of csrA rpoS mutant strains was very poor on acetate as a sole carbon source. Surprisingly, growth also was inhibited specifically by the addition of modest amounts of acetate to rich media (e.g., tryptone broth). Cultures grown in the presence of >/=25 mM acetate consisted substantially of glycogen biosynthesis (glg) mutants, which were no longer inhibited by acetate. Several classes of glg mutations were mapped to known and novel loci. Several hypotheses were examined to provide further insight into the effects of acetate on growth and metabolism in these strains. We determined that csrA positively regulates acs (acetyl coenzyme A synthetase; Acs) expression and isocitrate lyase activity without affecting key TCA cycle enzymes or phosphotransacetylase. TCA cycle intermediates or pyruvate, but not glucose, galactose, or glycerol, restored growth and prevented the glg mutations in the presence of acetate. Furthermore, amino acid uptake was inhibited by acetate specifically in the csrA rpoS strain. We conclude that central carbon flux imbalance, inhibition of amino acid uptake, and a deficiency in acetate metabolism apparently are combined to cause metabolic stress by depleting the TCA cycle. PMID- 10692370 TI - Substrate specificity of naphthalene dioxygenase: effect of specific amino acids at the active site of the enzyme. AB - The three-component naphthalene dioxygenase (NDO) enzyme system carries out the first step in the aerobic degradation of naphthalene by Pseudomonas sp. strain NCIB 9816-4. The three-dimensional structure of NDO revealed that several of the amino acids at the active site of the oxygenase are hydrophobic, which is consistent with the enzyme's preference for aromatic hydrocarbon substrates. Although NDO catalyzes cis-dihydroxylation of a wide range of substrates, it is highly regio- and enantioselective. Site-directed mutagenesis was used to determine the contributions of several active-site residues to these aspects of catalysis. Amino acid substitutions at Asn-201, Phe-202, Val-260, Trp-316, Thr 351, Trp-358, and Met-366 had little or no effect on product formation with naphthalene or biphenyl as substrates and had slight but significant effects on product formation from phenanthrene. Amino acid substitutions at Phe-352 resulted in the formation of cis-naphthalene dihydrodiol with altered stereochemistry [92 to 96% (+)-1R,2S], compared to the enantiomerically pure [>99% (+)-1R,2S] product formed by the wild-type enzyme. Substitutions at position 352 changed the site of oxidation of biphenyl and phenanthrene. Substitution of alanine for Asp-362, a ligand to the active-site iron, resulted in a completely inactive enzyme. PMID- 10692371 TI - Penicillin-binding protein-related factor A is required for proper chromosome segregation in Bacillus subtilis. AB - Previous work has shown that the ponA gene, encoding penicillin-binding protein 1 (PBP1), is in a two-gene operon with prfA (PBP-related factor A) (also called recU), which encodes a putative 206-residue basic protein (pI = 10.1) with no significant sequence homology to proteins with known functions. Inactivation of prfA results in cells that grow slower and vary significantly in length relative to wild-type cells. We now show that prfA mutant cells have a defect in chromosome segregation resulting in the production of approximately 0.9 to 3% anucleate cells in prfA cultures grown at 30 or 37 degrees C in rich medium and that the lack of PrfA exacerbates the chromosome segregation defect in smc and spoOJ mutant cells. In addition, overexpression of prfA was found to be toxic for and cause nucleoid condensation in Escherichia coli. PMID- 10692372 TI - Analysis of Escherichia coli RecA interactions with LexA, lambda CI, and UmuD by site-directed mutagenesis of recA. AB - An early event in the induction of the SOS system of Escherichia coli is RecA mediated cleavage of the LexA repressor. RecA acts indirectly as a coprotease to stimulate repressor self-cleavage, presumably by forming a complex with LexA. How complex formation leads to cleavage is not known. As an approach to this question, it would be desirable to identify the protein-protein interaction sites on each protein. It was previously proposed that LexA and other cleavable substrates, such as phage lambda CI repressor and E. coli UmuD, bind to a cleft located between two RecA monomers in the crystal structure. To test this model, and to map the interface between RecA and its substrates, we carried out alanine scanning mutagenesis of RecA. Twenty double mutations were made, and cells carrying them were characterized for RecA-dependent repair functions and for coprotease activity towards LexA, lambda CI, and UmuD. One mutation in the cleft region had partial defects in cleavage of CI and (as expected from previous data) of UmuD. Two mutations in the cleft region conferred constitutive cleavage towards CI but not towards LexA or UmuD. By contrast, no mutations in the cleft region or elsewhere in RecA were found to specifically impair the cleavage of LexA. Our data are consistent with binding of CI and UmuD to the cleft between two RecA monomers but do not provide support for the model in which LexA binds in this cleft. PMID- 10692373 TI - Transposon insertions in the Flavobacterium johnsoniae ftsX gene disrupt gliding motility and cell division. AB - Flavobacterium johnsoniae is a gram-negative bacterium that exhibits gliding motility. To determine the mechanism of flavobacterial gliding motility, we isolated 33 nongliding mutants by Tn4351 mutagenesis. Seventeen of these mutants exhibited filamentous cell morphology. The region of DNA surrounding the transposon insertion in the filamentous mutant CJ101-207 was cloned and sequenced. The transposon was inserted in a gene that was similar to Escherichia coli ftsX. Two of the remaining 16 filamentous mutants also carried insertions in ftsX. Introduction of the wild-type F. johnsoniae ftsX gene restored motility and normal cell morphology to each of the three ftsX mutants. CJ101-207 appears to be blocked at a late stage of cell division, since the filaments produced cross walls but cells failed to separate. In E. coli, FtsX is thought to function with FtsE in translocating proteins involved in potassium transport, and perhaps proteins involved in cell division, into the cytoplasmic membrane. Mutations in F. johnsoniae ftsX may prevent translocation of proteins involved in cell division and proteins involved in gliding motility into the cytoplasmic membrane, thus resulting in defects in both processes. Alternatively, the loss of gliding motility may be an indirect result of the defect in cell division. The inability to complete cell division may alter the cell architecture and disrupt gliding motility by preventing the synthesis, assembly, or functioning of the motility apparatus. PMID- 10692375 TI - Role of cell shape in determination of the division plane in Schizosaccharomyces pombe: random orientation of septa in spherical cells. AB - The establishment of growth polarity in Schizosaccharomyces pombe cells is a combined function of the cytoplasmic cytoskeleton and the shape of the cell wall inherited from the mother cell. The septum that divides the cylindrical cell into two siblings is formed midway between the growing poles and perpendicularly to the axis that connects them. Since the daughter cells also extend at their ends and form their septa at right angles to the longitudinal axis, their septal (division) planes lie parallel to those of the mother cell. To gain a better understanding of how this regularity is ensured, we investigated septation in spherical cells that do not inherit morphologically predetermined cell ends to establish poles for growth. We studied four mutants (defining four novel genes), over 95% of whose cells displayed a completely spherical morphology and a deficiency in mating and showed a random distribution of cytoplasmic microtubules, Tea1p, and F-actin, indicating that the cytoplasmic cytoskeleton was poorly polarized or apolar. Septum positioning was examined by visualizing septa and division scars by calcofluor staining and by the analysis of electron microscopic images. Freeze-substitution, freeze-etching, and scanning electron microscopy were used. We found that the elongated bipolar shape is not essential for the determination of a division plane that can separate the postmitotic nuclei. However, it seems to be necessary for the maintenance of the parallel orientation of septa over the generations. In the spherical cells, the division scars and septa usually lie at angles to each other on the cell surface. We hypothesize that the shape of the cell indirectly affects the positioning of the septum by directing the extension of the spindle. PMID- 10692374 TI - Biochemical and physical properties of the Methanococcus jannaschii 20S proteasome and PAN, a homolog of the ATPase (Rpt) subunits of the eucaryal 26S proteasome. AB - The 20S proteasome is a self-compartmentalized protease which degrades unfolded polypeptides and has been purified from eucaryotes, gram-positive actinomycetes, and archaea. Energy-dependent complexes, such as the 19S cap of the eucaryal 26S proteasome, are assumed to be responsible for the recognition and/or unfolding of substrate proteins which are then translocated into the central chamber of the 20S proteasome and hydrolyzed to polypeptide products of 3 to 30 residues. All archaeal genomes which have been sequenced are predicted to encode proteins with up to approximately 50% identity to the six ATPase subunits of the 19S cap. In this study, one of these archaeal homologs which has been named PAN for proteasome-activating nucleotidase was characterized from the hyperthermophile Methanococcus jannaschii. In addition, the M. jannaschii 20S proteasome was purified as a 700-kDa complex by in vitro assembly of the alpha and beta subunits and has an unusually high rate of peptide and unfolded-polypeptide hydrolysis at 100 degrees C. The 550-kDa PAN complex was required for CTP- or ATP-dependent degradation of beta-casein by archaeal 20S proteasomes. A 500-kDa complex of PAN(Delta1-73), which has a deletion of residues 1 to 73 of the deduced protein and disrupts the predicted N-terminal coiled-coil, also facilitated this energy dependent proteolysis. However, this deletion increased the types of nucleotides hydrolyzed to include not only ATP and CTP but also ITP, GTP, TTP, and UTP. The temperature optimum for nucleotide (ATP) hydrolysis was reduced from 80 degrees C for the full-length protein to 65 degrees C for PAN(Delta1-73). Both PAN protein complexes were stable in the absence of ATP and were inhibited by N ethylmaleimide and p-chloromercuriphenyl-sulfonic acid. Kinetic analysis reveals that the PAN protein has a relatively high V(max) for ATP and CTP hydrolysis of 3.5 and 5.8 micromol of P(i) per min per mg of protein as well as a relatively low affinity for CTP and ATP with K(m) values of 307 and 497 microM compared to other proteins of the AAA family. Based on electron micrographs, PAN and PAN(Delta1-73) apparently associate with the ends of the 20S proteasome cylinder. These results suggest that the M. jannaschii as well as related archaeal 20S proteasomes require a nucleotidase complex such as PAN to mediate the energy dependent hydrolysis of folded-substrate proteins and that the N-terminal 73 amino acid residues of PAN are not absolutely required for this reaction. PMID- 10692376 TI - Dual roles of Bradyrhizobium japonicum nickelin protein in nickel storage and GTP dependent Ni mobilization. AB - The hydrogenase accessory protein HypB, or nickelin, has two functions in the N(2)-fixing, H(2)-oxidizing bacterium Bradyrhizobium japonicum. One function of HypB involves the mobilization of nickel into hydrogenase. HypB also carries out a nickel storage/sequestering function in B. japonicum, binding nine nickel ions per monomer. Here we report that the two roles (nickel mobilization and storage) of HypB can be separated in vitro and in vivo using molecular and biochemical approaches. The role of HypB in hydrogenase maturation is completely dependent on its intrinsic GTPase activity; strains which produce a HypB protein that is severely deficient in GTPase activity but that fully retains nickel-sequestering ability cannot produce active hydrogenase even upon prolonged nickel supplementation. A HypB protein that lacks the nickel-binding polyhistidine region near the N terminus lacks only the nickel storage capacity function; it is still able to bind a single nickel ion and also retains complete GTPase activity. PMID- 10692377 TI - Identification of genes in the RosR regulon of Rhizobium etli. AB - RosR is a determinant of nodulation competitiveness and cell surface characteristics of Rhizobium etli and has sequence similarity to a family of transcriptional repressors. To understand how RosR affects these phenotypes, we mutagenized a rosR mutant derivative of R. etli strain CE3 with a mini-Tn5 that contains a promoterless gusA gene at one end, which acts as a transcriptional reporter. Using a mass-mating technique, we introduced rosR into each mutant in trans and screened for mutants that expressed different levels of beta glucuronidase activity in the presence and absence of rosR. A screen of 18,000 mutants identified 52 insertions in genes negatively regulated by RosR and 1 insertion in a gene positively regulated by RosR. Nucleotide sequence analysis of the regions flanking the insertions suggests that RosR regulates genes of diverse function, including those involved in polysaccharide production and in carbohydrate metabolism and those in a region containing sequence similarity to virC1 and virD3 from Agrobacterium tumefaciens. Two of the mutants produced colonies with altered morphology and were more competitive in nodulation than was CE3DeltarosR, the rosR parent. One mutant that contained an insertion in a gene with similarity to exsH of Sinorhizobium meliloti did not nodulate the plant host Phaseolus vulgaris without rosR. These results indicate that RosR directly or indirectly influences expression of diverse genes in R. etli, some of which affect the cell surface and nodulation competitiveness. PMID- 10692378 TI - Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli. AB - Although general physiological functions have been ascribed to the high-molecular weight penicillin binding proteins (PBPs) of Escherichia coli, the low-molecular weight PBPs have no well-defined biological roles. When we examined the morphology of a set of E. coli mutants lacking multiple PBPs, we observed that strains expressing active PBP 5 produced cells of normal shape, while mutants lacking PBP 5 produced cells with altered diameters, contours, and topological features. These morphological effects were visible in untreated cells, but the defects were exacerbated in cells forced to filament by inactivation of PBP 3 or FtsZ. After filamentation, cellular diameter varied erratically along the length of individual filaments and many filaments exhibited extensive branching. Also, in general, the mean diameter of cells lacking PBP 5 was significantly increased compared to that of cells from isogenic strains expressing active PBP 5. Expression of cloned PBP 5 reversed the effects observed in DeltadacA mutants. Although deletion of PBP 5 was required for these phenotypes, the absence of additional PBPs magnified the effects. The greatest morphological alterations required that at least three PBPs in addition to PBP 5 be deleted from a single strain. In the extreme cases in which six or seven PBPs were deleted from a single mutant, cells and cell filaments expressing PBP 5 retained a normal morphology but cells and filaments lacking PBP 5 were aberrant. In no case did mutation of another PBP produce the same drastic morphological effects. We conclude that among the low-molecular-weight PBPs, PBP 5 plays a principle role in determining cell diameter, surface uniformity, and overall topology of the peptidoglycan sacculus. PMID- 10692379 TI - Membrane topology of the NixA nickel transporter of Helicobacter pylori: two nickel transport-specific motifs within transmembrane helices II and III. AB - NixA, the high-affinity cytoplasmic membrane nickel transport protein of Helicobacter pylori, imports Ni(2+) into the cell for insertion into the active site of the urease metalloenzyme, which is required for gastric colonization. NixA fractionates with the cytoplasmic membrane, and protein cross-linking studies suggest that NixA functions as a monomer. A preliminary topological model of NixA with seven transmembrane domains was previously proposed based on hydropathy, charge dispersion, and homology to other transporters. To test the proposed topology of NixA and relate critical residues to specific structural elements, a series of 21 NixA-LacZ and 21 NixA-PhoA fusions were created along the entire length of the protein. Expression of reporter fusions was confirmed by Western blotting with beta-galactosidase- and alkaline phosphatase-specific antisera. The activities of reporter fusions near to and upstream of the predicted translational initiation demonstrated the presence of an additional amino-terminal transmembrane domain including a membrane localization signal. Activities of fusions immediately adjacent to motifs which have been shown to be requisite for Ni(2+) transport localized these motifs entirely within transmembrane domains II and III. Fusion activities localized six additional Asp and Glu residues which reduced Ni(2+) transport by >90% when mutated within or immediately adjacent to transmembrane domains II, V, VI, and VII. All fusions strongly support a model of NixA in which the amino and carboxy termini are located in the cytoplasm and the protein possesses eight transmembrane domains. PMID- 10692380 TI - Vibrio cholerae VibF is required for vibriobactin synthesis and is a member of the family of nonribosomal peptide synthetases. AB - A 7.5-kbp fragment of chromosomal DNA downstream of the Vibrio cholerae vibriobactin outer membrane receptor, viuA, and the vibriobactin utilization gene, viuB, was recovered from a Sau3A lambda library of O395 chromosomal DNA. By analogy with the genetic organization of the Escherichia coli enterobactin gene cluster, in which the enterobactin biosynthetic and transport genes lie adjacent to the enterobactin outer membrane receptor, fepA, and the utilization gene, fes, the cloned DNA was examined for the ability to restore siderophore synthesis to E. coli ent mutants. Cross-feeding studies demonstrated that an E. coli entF mutant complemented with the cloned DNA regained the ability to synthesize enterobactin and to grow in low-iron medium. Sequence analysis of the cloned chromosomal DNA revealed an open reading frame downstream of viuB which encoded a deduced protein of greater than 2,158 amino acids, homologous to Yersinia sp. HMWP2, Vibrio anguillarum AngR, and E. coli EntF. A mutant with an in-frame deletion of this gene, named vibF, was created with classical V. cholerae strain O395 by in vivo marker exchange. In cross-feeding studies, this mutant was unable to synthesize ferric vibriobactin but was able to utilize exogenous siderophore. Complementation of the mutant with a cloned vibF fragment restored vibriobactin synthesis to normal. The expression of the vibF promoter was found to be negatively regulated by iron at the transcriptional level, under the control of the V. cholerae fur gene. Expression of vibF was not autoregulatory and neither affected nor was affected by the expression of irgA or viuA. The promoter of vibF was located by primer extension and was found to contain a dyad symmetric nucleotide sequence highly homologous to the E. coli Fur binding consensus sequence. A footprint of purified V. cholerae Fur on the vibF promoter, overlapping the Fur binding consensus sequence, was observed using DNase I footprinting. The protein product of vibF is homologous to the multifunctional nonribosomal protein synthetases and is necessary for the biosynthesis of vibriobactin. PMID- 10692381 TI - CTXphi infection of Vibrio cholerae requires the tolQRA gene products. AB - CTXphi is a lysogenic filamentous bacteriophage that encodes cholera toxin. Filamentous phages that infect Escherichia coli require both a pilus and the products of tolQRA in order to enter host cells. We have previously shown that toxin-coregulated pilus (TCP), a type IV pilus that is an essential Vibrio cholerae intestinal colonization factor, serves as a receptor for CTXphi. To test whether CTXphi also depends upon tol gene products to infect V. cholerae, we identified and inactivated the V. cholerae tolQRAB orthologues. The predicted amino acid sequences of V. cholerae TolQ, TolR, TolA, and TolB showed significant similarity to the corresponding E. coli sequences. V. cholerae strains with insertion mutations in tolQ, tolR, or tolA were reduced in their efficiency of CTXphi uptake by 4 orders of magnitude, whereas a strain with an insertion mutation in tolB showed no reduction in CTXphi entry. We could detect CTXphi infection of TCP(-) V. cholerae, albeit at very low frequencies. However, strains with mutations in both tcpA and either tolQ, tolR, or tolA were completely resistant to CTXphi infection. Thus, CTXphi, like the E. coli filamentous phages, uses both a pilus and TolQRA to enter its host. This suggests that the pathway for filamentous phage entry into cells is conserved between host bacterial species. PMID- 10692382 TI - Glutathione is involved in environmental stress responses in Rhizobium tropici, including acid tolerance. AB - The isolation of rhizobial strains which exhibit an intrinsic tolerance to acidic conditions has been reported and has facilitated studies on the basic mechanisms underlying acid tolerance. Rhizobium tropici strain CIAT899 displays a high intrinsic tolerance to acidity and therefore was used in this work to study the molecular basis of bacterial responses to acid conditions and other environmental stresses. We generated a collection of R. tropici CIAT899 mutants affected in acid tolerance using Tn5-luxAB mutagenesis, and one mutant strain (CIAT899-13T2), which fails to grow under acid conditions, was characterized in detail. Strain CIAT899-13T2 was found to contain a single Tn5-luxAB insertion in a gene showing a high degree of similarity with the Escherichia coli gshB gene, encoding the enzyme glutathione synthetase. Intracellular potassium pools and intracellular pH levels were found to be lower in the mutant than in the parent. The glutathione deficient mutant was shown to be sensitive to weak organic acids, osmotic and oxidative stresses, and the presence of methylglyoxal. Glutathione restores responses to these stresses almost to wild-type levels. Our data show that in R. tropici the production of glutathione is essential for growth in extreme environmental conditions. The mutant strain CIAT899-13T2 induced effective nodules; however, it was found to be outcompeted by the wild-type strain in coinoculation experiments. PMID- 10692383 TI - AcrD of Escherichia coli is an aminoglycoside efflux pump. AB - AcrD, a transporter belonging to the resistance-nodulation-division family, was shown to participate in the efflux of aminoglycosides. Deletion of the acrD gene decreased the MICs of amikacin, gentamicin, neomycin, kanamycin, and tobramycin by a factor of two to eight, and DeltaacrD cells accumulated higher levels of [(3)H]dihydrostreptomycin and [(3)H]gentamicin than did the parent strain. PMID- 10692384 TI - Isolation of an inner membrane-derived subfraction that supports in vitro replication of a mini-RK2 plasmid in Escherichia coli. AB - Previous results have demonstrated that the inner, but not the outer, membrane fraction of Escherichia coli is the site of membrane-associated DNA replication of plasmid RK2, a broad-host-range plasmid capable of replication in a wide variety of gram-negative hosts (K. Michaels, J. Mei, and W. Firshein, Plasmid 32:19-31, 1994). To resolve the inner membrane replication site further, the procedure of Ishidate et al. (K. Ishidate, E. S. Creeger, J. Zrike, S. Deb, G. Glauner, T. J. MacAlister, and L. I. Rothfield, J. Biol. Chem. 261:428-443, 1986) was used to separate the inner membrane into a number of subfractions, of which only one, a small subfraction containing only 10% of the entire membrane, was found to synthesize DNA inhibited by antibody prepared against the plasmid encoded initiation protein TrfA. This is the same subfraction that was also found to bind oriV and TrfA to the greatest extent in filter binding assays (J. Mei, S. Benashski, and W. Firshein, J. Bacteriol. 177:6766-6772, 1995). PMID- 10692385 TI - Activation of SoxR by overproduction of desulfoferrodoxin: multiple ways to induce the soxRS regulon. AB - The soxRS response, which protects cells against superoxide toxicity, is triggered by the oxidation of SoxR, a transcription factor. Superoxide excess and NADPH depletion induce the regulon. Unexpectedly, we found that the overproduction of desulfoferrodoxin, a superoxide reductase from sulfate-reducing bacteria, also induced this response. We suggest that desulfoferrodoxin interferes with the reducing pathway that keeps SoxR in its inactive form. PMID- 10692386 TI - Nitrate assimilation genes of the marine diazotrophic, filamentous cyanobacterium Trichodesmium sp. strain WH9601. AB - A 4.0-kb DNA fragment of Trichodesmium sp. strain WH9601 contained gene sequences encoding the nitrate reduction enzymes, nirA and narB. A third gene positioned between nirA and narB encodes a putative membrane protein with similarity to the nitrate permeases of Bacillus subtilis (NasA) and Emericella nidulans (CrnA). The gene was shown to functionally complement a DeltanasA mutant of B. subtilis and was assigned the name napA (nitrate permease). NapA was involved in both nitrate and nitrite uptake by the complemented B. subtilis cells. napA is distinct from the nrt genes that encode the nitrate transporter of freshwater cyanobacteria. PMID- 10692387 TI - Membrane association of the Escherichia coli enterobactin synthase proteins EntB/G, EntE, and EntF. AB - The cytosolic proteins EntE, EntF, and EntB/G, which are Escherichia coli enzymes necessary for the final stage of enterobactin synthesis, are released by osmotic shock. Here, consistent with the idea that cytoplasmic proteins found in shockates have an affinity for membranes, a small fraction of each was found in membrane preparations. Two procedures demonstrated that the enzymes were enriched in a minor membrane fraction of buoyant density intermediate between that of cytoplasmic and outer membranes, providing indirect support for the notion that these proteins have a role in enterobactin excretion as well as synthesis. PMID- 10692388 TI - The right end of the vir region of an octopine-type Ti plasmid contains four new members of the vir regulon that are not essential for pathogenesis. AB - We sequenced the virD-virE, virE-virF, and virF-T-DNA intergenic regions of an octopine Ti plasmid. Four newly described genes were induced by the vir gene inducer acetosyringone, two of which are conserved in the nopaline-type Ti plasmid pTiC58. One gene resembles a family of phosphatase genes. Each of these genes is dispensable for tumorigenesis. PMID- 10692389 TI - Mechanism and cellular applications of a green fluorescent protein-based halide sensor. AB - We report the application of a targetable green fluorescent protein-based cellular halide indicator. Fluorescence titrations of the purified recombinant yellow fluorescent protein YFP-H148Q indicated a pK(a) of 7.14 in the absence of Cl(-), which increased to 7.86 at 150 mM Cl(-). At pH 7.5, YFP-H148Q fluorescence decreased maximally by approximately 2-fold with a K(D) of 100 mM Cl(-). YFP H148Q had a fluorescence lifetime of 3.1 ns that was independent of pH and [Cl( )]. Circular dichroism and absorption spectroscopy revealed distinct Cl(-) dependent spectral changes indicating Cl(-)/YFP binding. Stopped-flow kinetic analysis showed a biexponential time course of YFP-H148Q fluorescence (time constants <100 ms) in response to changes in pH or [Cl(-)], establishing a 1:1 YFP-H148Q/Cl(-) binding mechanism. Photobleaching analysis revealed a millisecond triplet state relaxation process that was insensitive to anions and aqueous-phase quenchers. The anion selectivity sequence for YFP-H148Q quenching (ClO(4)(-) approximately I(-) > SCN(-) > NO(3)(-) > Cl(-) > Br(-) > formate > acetate) indicated strong binding of weakly hydrated chaotropic ions. The biophysical data suggest that YFP-H148Q anion sensitivity involves ground state anion binding to a site close to the tri-amino acid chromophore. YFP-H148Q transfected mammalian cells were brightly fluorescent with cytoplasmic/nuclear staining. Ionophore calibrations indicated similar YFP-H148Q pH and anion sensitivities in cells and aqueous solutions. Cyclic AMP-regulated Cl(-) transport through plasma membrane cystic fibrosis transmembrane conductance regulator Cl(-) channels was assayed with excellent sensitivity from the time course of YFP-H148Q fluorescence in response to extracellular Cl(-)/I(-) exchange. The green fluorescent protein based halide sensor described here should have numerous applications, such as anion channel cloning by screening of mammalian expression libraries and discovery of compounds that correct the cystic fibrosis phenotype by screening of combinatorial libraries. PMID- 10692390 TI - p53 regulates the expression of the tumor suppressor gene maspin. AB - Maspin has been shown to inhibit tumor cell invasion and metastasis in breast tumor cells. Maspin expression was detected in normal breast and prostate epithelial cells, whereas tumor cells exhibited reduced or no expression. However, the regulatory mechanism of maspin expression remains unknown. We report here a rapid and robust induction of maspin expression in prostate cancer cells (LNCaP, DU145, and PC3) and breast tumor cells (MCF7) following wild type p53 expression from an adenovirus p53 expression vector (AdWTp53). p53 activates the maspin promoter by binding directly to the p53 consensus-binding site present in the maspin promoter. DNA-damaging agents and cytotoxic drugs induced endogenous maspin expression in cells containing the wild type p53. Maspin expression was refractory to the DNA-damaging agents in cells containing mutant p53. These results, combined with recent studies of the tumor metastasis suppressor gene KAI1 and plasminogen activator inhibitor 1 (PAI1), define a new category of molecular targets of p53 that have the potential to negatively regulate tumor invasion and/or metastasis. PMID- 10692391 TI - Phosphorylation of a Src kinase at the autophosphorylation site in the absence of Src kinase activity. AB - Exposure of cells to oxidants increases the phosphorylation of the Src family tyrosine protein kinase Lck at Tyr-394, a conserved residue in the activation loop of the catalytic domain. Kinase-deficient Lck expressed in fibroblasts that do not express any endogenous Lck has been shown to be phosphorylated at Tyr-394 following H(2)O(2) treatment to an extent indistinguishable from that seen with wild type Lck. This finding indicates that a kinase other than Lck itself is capable of phosphorylating Tyr-394. Because fibroblasts express other Src family members, it remained to be determined whether the phosphorylation of Tyr-394 was carried out by another Src family kinase or by an unrelated tyrosine protein kinase. We examined here whether Tyr-394 in kinase-deficient Lck was phosphorylated following exposure of cells devoid of endogenous Src family kinase activity to H(2)O(2). Strikingly, treatment of such cells with H(2)O(2) led to the phosphorylation of Tyr-394 to an extent identical to that seen with wild type Lck, demonstrating that Src family kinases are not required for H(2)O(2)-induced phosphorylation of Lck. Furthermore, this efficient phosphorylation of Lck at Tyr 394 in non-lymphoid cells suggests the existence of an ubiquitous activator of Src family kinases. PMID- 10692392 TI - VRAP is an adaptor protein that binds KDR, a receptor for vascular endothelial cell growth factor. AB - A protein that binds the intracellular domain of KDR (KDR-IC), a receptor for vascular endothelial cell growth factor (VEGF), was identified by two-hybrid screening. Two-hybrid mapping showed that the VEGF receptor-associated protein (VRAP) interacted with tyrosine 951 in the kinase insert domain of KDR. Northern blot analysis identified multiple VRAP transcripts in peripheral leukocytes, spleen, thymus, heart, lung, and human umbilical vein endothelial cells (HUVEC). The predominant VRAP mRNA encodes a 389-amino acid protein that contains an SH2 domain and a C-terminal proline-rich motif. In HUVEC, VEGF promotes association of VRAP with KDR. Phospholipase C gamma and phosphatidylinositol 3-kinase, effector proteins that are downstream of KDR and important to VEGF-induced endothelial cell survival and proliferative responses, associate constitutively with VRAP. These observations identify VRAP as an adaptor that recruits cytoplasmic signaling proteins to KDR, which plays an important role in normal and pathological angiogenesis. PMID- 10692393 TI - Inositol tetrakisphosphate as a frequency regulator in calcium oscillations in HeLa cells. AB - Cellular signaling mediated by inositol (1,4,5)trisphosphate (Ins(1, 4,5)P(3)) results in oscillatory intracellular calcium (Ca(2+)) release. Because the amplitude of the Ca(2+) spikes is relatively invariant, the extent of the agonist mediated effects must reside in their ability to regulate the oscillating frequency. Using electroporation techniques, we show that Ins(1,4,5)P(3), Ins(1,3,4, 5)P(4), and Ins(1,3,4,6)P(4) cause a rapid intracellular Ca(2+) release in resting HeLa cells and a transient increase in the frequency of ongoing Ca(2+) oscillations stimulated by histamine. Two poorly metabolizable analogs of Ins(1,4,5)P(3), Ins(2,4,5)P(3), and 2,3-dideoxy-Ins(1,4,5)P(3), gave a single Ca(2+) spike and failed to alter the frequency of ongoing oscillations. Complete inhibition of Ins(1,4,5)P(3) 3-kinase (IP3K) by either adriamycin or its specific antibody blocked Ca(2+) oscillations. Partial inhibition of IP3K causes a significant reduction in frequency. Taken together, our results indicate that Ins(1,3,4,5)P(4) is the frequency regulator in vivo, and IP3K, which phosphorylates Ins(1,4, 5)P(3) to Ins(1,3,4,5)P(4), plays a major regulatory role in intracellular Ca(2+) oscillations. PMID- 10692394 TI - Apaf-1 oligomerizes into biologically active approximately 700-kDa and inactive approximately 1.4-MDa apoptosome complexes. AB - Apaf-1, by binding to and activating caspase-9, plays a critical role in apoptosis. Oligomerization of Apaf-1, in the presence of dATP and cytochrome c, is required for the activation of caspase-9 and produces a caspase activating apoptosome complex. Reconstitution studies with recombinant proteins have indicated that the size of this complex is very large in the order of approximately 1.4 MDa. We now demonstrate that dATP activation of cell lysates results in the formation of two large Apaf-1-containing apoptosome complexes with M(r) values of approximately 1.4 MDa and approximately 700 kDa. Kinetic analysis demonstrates that in vitro the approximately 700-kDa complex is produced more rapidly than the approximately 1.4 MDa complex and exhibits a much greater ability to activate effector caspases. Significantly, in human tumor monocytic cells undergoing apoptosis after treatment with either etoposide or N-tosyl-l phenylalanyl chloromethyl ketone (TPCK), the approximately 700-kDa Apaf-1 containing apoptosome complex was predominately formed. This complex processed effector caspases. Thus, the approximately 700-kDa complex appears to be the correctly formed and biologically active apoptosome complex, which is assembled during apoptosis. PMID- 10692395 TI - Human PC4 is a substrate-specific inhibitor of RNA polymerase II phosphorylation. AB - The activity of cyclin-dependent protein kinases (cdks) is physiologically regulated by phosphorylation, association with the specific cyclin subunits, and repression by specific cdk inhibitors. All three physiological regulatory mechanisms are specific for one or more cdks, but none is known to be substrate specific. In contrast, synthetic cdk peptide inhibitors that specifically inhibit cdk phosphorylation of only some substrates, "aptamers," have been described. Here, we show that PC4, a naturally occurring transcriptional coactivator, competitively inhibits cdk-1, -2, and -7-mediated phosphorylation of the largest subunit of RNA polymerase II (RNAPII), but it does not inhibit phosphorylation of other substrates of the same kinases. Interestingly, the phosphorylated form of PC4 is devoid of kinase inhibitory activity. We also show that wild-type PC4 but not the kinase inhibitory-deficient mutant of PC4 represses transcription in vivo. Our results point to a novel role for PC4 as a specific inhibitor of RNAPII phosphorylation. PMID- 10692396 TI - Smad6 is a Smad1/5-induced smad inhibitor. Characterization of bone morphogenetic protein-responsive element in the mouse Smad6 promoter. AB - Smad6 is an inhibitory Smad that is induced by bone morphogenetic proteins (BMPs) and interferes with BMP signaling. We have isolated the mouse Smad6 promoter and identified the regions responsible for transcriptional activation by BMPs. The proximal BMP-responsive element (PBE) in the Smad6 promoter is important for the transcriptional activation by BMPs and contains a 28-base pair GC-rich sequence including four overlapping copies of the GCCGnCGC-like motif, which is a binding site for Drosophila Mad and Medea. We generated a luciferase reporter construct (3GC2-Lux) containing three repeats of the GC-rich sequence derived from the PBE. BMPs and BMP receptors induced transcriptional activation of 3GC2-Lux in various cell types, and this activation was enhanced by cotransfection of BMP-responsive Smads, i.e. Smad1 or Smad5. Moreover, direct DNA binding of BMP-responsive Smads and common-partner Smad4 to the GC-rich sequence of PBE was observed. These results indicate that the expression of Smad6 is regulated by the effects of BMP activated Smad1/5 on the Smad6 promoter. PMID- 10692397 TI - Coordinate copper- and oxygen-responsive Cyc6 and Cpx1 expression in Chlamydomonas is mediated by the same element. AB - Chlamydomonas reinhardtii activates the transcription of the Cyc6 and the Cpx1 genes (encoding cytochrome c(6) and coprogen oxidase) in response to copper deficiency. Mutational analysis of promoter regions of the Cyc6 and Cpx1 genes revealed a four nucleotide sequence, GTAC, which was absolutely essential for copper responsiveness. The Cyc6 promoter contains two copper response elements, each with a functionally important GTAC sequence, whereas the Cpx1 promoter contains only one. This may contribute to the stronger and more tightly regulated expression of the Cyc6 gene. Mutation or deletion of sequences flanking the GTACs implicates additional nucleotides contributing to copper-responsive expression, but none are absolutely essential. Metal ion selectivity of Cpx1 expression is identical to that described previously for Cyc6 and is restricted to the copper deficiency-induced Cpx1 transcript. The Cyc6 and Cpx1 genes are also induced by oxygen deficiency. Reporter gene constructs indicate that the induction occurs at the level of transcription and requires the same GTAC sequence that is critical for copper responsiveness. We suggest that components of the copper-responsive signal transduction pathway are used for some of the changes in gene expression in hypoxic cells. PMID- 10692398 TI - Molecular and functional properties of the human alpha(1G) subunit that forms T type calcium channels. AB - We describe here several novel properties of the human alpha(1G) subunit that forms T-type calcium channels. The partial intron/exon structure of the corresponding gene CACNA1G was defined and several alpha(1G) isoforms were identified, especially two isoforms that exhibit a distinct III-IV loop: alpha(1G a) and alpha(1G-b). Northern blot and dot blot analyses indicated that alpha(1G) mRNA is predominantly expressed in the brain, especially in thalamus, cerebellum, and substantia nigra. Additional experiments have also provided evidence that alpha(1G) mRNA is expressed at a higher level during fetal life in nonneuronal tissues (i.e. kidney, heart, and lung). Functional expression in HEK 293 cells of a full-length cDNA encoding the shortest alpha(1G) isoform identified to date, alpha(1G-b), resulted in transient, low threshold activated Ca(2+) currents with the expected permeability ratio (I(Sr) > I(Ca) >/= I(Ba)) and channel conductance ( approximately 7 pS). These properties, together with slowly deactivating tail currents, are typical of those of native T-type Ca(2+) channels. This alpha(1G) related current was inhibited by mibefradil (IC(50) = 2 microM) and weakly blocked by Ni(2+) ions (IC(50) = 148 microM) and amiloride (IC(50) > 1 mM). We showed that steady state activation and inactivation properties of this current can generate a "window current" in the range of -65 to -55 mV. Using neuronal action potential waveforms, we show that alpha(1G) channels produce a massive and sustained Ca(2+) influx due to their slow deactivation properties. These latter properties would account for the specificity of Ca(2+) influx via T-type channels that occurs in the range of physiological resting membrane potentials, differing considerably from the behavior of other Ca(2+) channels. PMID- 10692399 TI - Evidence for interactions between helices 5 and 8 and a role for the interdomain loop in tetracycline resistance mediated by hybrid Tet proteins. AB - An interdomain hybrid Tet protein consisting of a class C alpha domain and a class B beta domain (Tet(C/B)) lacks detectable efflux ability and provides only minimal levels of resistance to tetracycline (Tc) (3 microg/ml) compared with intact class B (256 microg/ml) and class C (64 microg/ml). Twenty-one independently isolated mutants of the Tet(C/B) protein with increased Tc resistance were generated by random chemical mutagenesis. Nine mutants with a Glu substitution for Gly-152 in helix 5 of the class C alpha domain produced a resistance of 48 microg/ml, whereas another 9 with an Asp replacement of Gly-247 in helix 8 of the class B beta domain mediated resistance at 32 microg/ml. The third type of mutation, found in 3 mutants expressing 24 microg/ml resistance, was a S202F replacement in the putative interdomain cytoplasmic loop of Tet(C/B). The latter underscores a previously unappreciated function of the interdomain cytoplasmic loop. All three types of Tet(C/B) mutant proteins were expressed in amounts comparable with that of the original protein and demonstrated restored energy-dependent efflux of tetracycline. Site-directed mutational analysis demonstrated that a Gly-247 to Asn mutation could also facilitate Tc resistance by the Tet(C/B) hybrid, and a negatively charged side chain at position 152 was required for Tet(C/B) activity. These mutations appear to promote the necessary functional interactions between the interclass domains that do not occur in the Tet(C/B) hybrid protein and suggest a direct association between helix 5 and helix 8 in the function of Tet efflux proteins. PMID- 10692400 TI - Identification of a key region of kinin B(1) receptor for high affinity binding of peptide antagonists. AB - To investigate the molecular basis for the specificity of ligand recognition in human kinin B(1) (B(1)R) and B(2) (B(2)R) receptors, we constructed a series of chimeric receptors by progressively replacing, from the N to the C terminus, the human B(2)R domains by their B(1) counterparts. The chimeric construct possessing the C-terminal tail and the transmembrane domain VII (TM VII) of the B(2)R (construct 6) displayed 7- and 20- fold decreased affinities for the B(1) agonist [(3)H]desArg(10)-kallidin (desArg(10)-KD) and the B(1) antagonist [(3)H]desArg(10)-[Leu(9)]-KD respectively, as compared with the wild-type B(1)R. Moreover, the substitution of the B(1) TM VII by its B(2) homologue TM increased the affinity for the pseudopeptide antagonists, Hoe140 and NPC 567. High affinity for desArg(10)-KD binding was fully regained when the B(2) residue Thr(287) was replaced in construct 6 by the corresponding B(1) Leu(294) residue. When the B(2) residue Tyr(295) was exchanged with the corresponding B(1) Phe(302), high affinity binding for both agonist and antagonist was recovered. Moreover, the L294T and F302Y mutant B(1)R exhibited 69- and 6.5-fold increases, respectively, in their affinities for the B(2) receptor antagonist, Hoe140. Therefore we proposed that Leu(294) and Phe(302) residues, which may not be directly involved in the binding of B(1)R ligands and, hence, their Thr(287) and Tyr(295) B(2) counterparts, are localized in a receptor region, which plays a pivotal role in the binding selectivity of the peptide or pseudopeptide kinin ligands. PMID- 10692401 TI - The multifunctional character of a geminivirus replication protein is reflected by its complex oligomerization properties. AB - Tomato golden mosaic virus (TGMV), a member of the geminivirus family, encodes one essential replication protein, AL1, and recruits the rest of the DNA replication apparatus from its plant host. TGMV AL1 is an oligomeric protein that binds double-stranded DNA and catalyzes cleavage and ligation of single-stranded DNA. The oligomerization domain, which is required for DNA binding, maps to a region that displays strong sequence and structural homology to other geminivirus Rep proteins. To assess the importance of conserved residues, we generated a series of site-directed mutations and analyzed their impact on AL1 function in vitro and in vivo. Two-hybrid experiments revealed that mutation of amino acids 157-159 inhibited AL1-AL1 interactions, whereas mutations at nearby residues reduced complex stability. Changes at positions 157-159 also disrupted interaction between the full-length mutant protein and a glutathione S transferase-AL1 oligomerization domain fusion in insect cells. The mutations had no detectable effect on oligomerization when both proteins contained full-length AL1 sequences, indicating that AL1 complexes can be stabilized by amino acids outside of the oligomerization domain. Nearly all of the oligomerization domain mutants were inhibited or severely attenuated in their ability to support AL1 directed viral DNA replication. In contrast, the same mutants were enhanced for AL1-mediated transcriptional repression. The replication-defective AL1 mutants also interfered with replication of a TGMV A DNA encoding wild type AL1. Full length mutant AL1 was more effective in the interference assays than truncated proteins containing the oligomerization domain. Together, these results suggested that different AL1 complexes mediate viral replication and transcriptional regulation and that replication interference involves multiple domains of the AL1 protein. PMID- 10692402 TI - Enhanced electron flux and reduced calmodulin dissociation may explain "calcium independent" eNOS activation by phosphorylation. AB - Bovine endothelial nitric oxide synthase (eNOS) is phosphorylated directly by the protein kinase Akt at serine 1179. Mutation of this residue to the negatively charged aspartate (S1179D eNOS) increases nitric oxide (NO) production constitutively, in the absence of agonist challenge. Here, we examine the potential mechanism of how aspartate at 1179 increases eNOS activity using purified proteins. Examination of NO production and cytochrome c reduction resulted in no substantial changes in the K(m)/EC(50) for L-arginine, calmodulin, and calcium, whereas there was a 2-fold increase in the rate of NO production for S1179D and a 2-4-fold increase in reductase activity (based on cytochrome c reduction). The observed increase in activity for both assays of NOS function indicates that a faster rate of electron flux through the reductase domain is likely the rate-limiting step in NO formation from eNOS. In addition, S1179D eNOS did show an increased resistance to inactivation by EGTA compared with wild type eNOS. These results suggest that a negative charge imposed at serine 1179, either by phosphorylation or by replacement with aspartate, increases eNOS catalytic activity by increasing electron flux at the reductase domain and by reducing calmodulin dissociation from activated eNOS when calcium levels are low. PMID- 10692403 TI - Free [ADP] and aerobic muscle work follow at least second order kinetics in rat gastrocnemius in vivo. AB - The relationship between free cytosolic [ADP] (and [P(i)]) and steady-state aerobic muscle work in rat gastrocnemius muscle in vivo using (31)P NMR was investigated. Anesthetized rats were ventilated and placed in a custom-built cradle fitted with a force transducer that could be placed into a 7-tesla NMR magnet. Muscle work was induced by supramaximal sciatic nerve stimulation that activated all fibers. Muscles were stimulated at 0.1, 0.2, 0.3, 0.4, 0.5, 0.8, 1.0, and 2.0 Hz until twitch force, phosphocreatine, and P(i) were unchanged between two consecutive spectra acquired in 4-min blocks (8-12 min). Parallel bench experiments were performed to measure total tissue glycogen, lactate, total creatine, and pyruvate in freeze-clamped muscles after 10 min of stimulation at each frequency. Up to 0.5 Hz, there was no significant change in muscle glycogen, lactate, and the lactate/pyruvate ratios between 8-12 min. At 0.8 Hz, there was a 17% fall in glycogen and a 65% rise in the muscle lactate with a concomitant fall in pH. Above this frequency, glycogen fell rapidly, lactate continued to rise, and ATP and pH declined. On the basis of these force and metabolic measurements, we estimated the maximal mitochondrial capacity (V(max)) to be 0.8 Hz. Free [ADP] was then calculated at each submaximal workload from measuring all the reactants of the creatine kinase equilibrium after adjusting the K'(CK) to the muscle temp (30 degrees C), pH, and pMg. We show that ADP (and P(i)) and tension-time integral follow a Hill relationship with at least a second order function. The K(0.5) values for free [ADP] and [P(i)] were 48 microM and 9 mM, respectively. Our data did not fit any form of the Michaelis-Menten equation. We therefore conclude that free cytosolic [ADP] and [P(i)] could potentially control steady state oxidative phosphorylation in skeletal muscle in vivo. PMID- 10692404 TI - Regulation of cloned cardiac L-type calcium channels by cGMP-dependent protein kinase. AB - We have studied the effect of 8-bromo-cyclic GMP (8-Br-cGMP) on cloned cardiac L type calcium channel currents to determine the site and mechanism of action underlying the functional effect. Rabbit cardiac alpha(1C) subunit, in the presence or absence of beta(1) subunit (rabbit skeletal muscle) or beta(2) subunit (rat cardiac/brain), was expressed in Xenopus oocytes, and two-electrode voltage-clamp recordings were made 2 or 3 days later. Application of 8-Br-cGMP caused decreases in calcium channel currents in cells expressing the alpha(1C) subunit, whether or not a beta subunit was co-expressed. No inhibition of currents by 8-Br-cGMP was observed in the presence of the protein kinase G inhibitor KT5823. Substitutions of serine residues by alanine were made at residues Ser(533) and Ser(1371) on the alpha(1C) subunit. As for wild type, the mutant S1371A exhibited inhibition of calcium channel currents by 8-Br-cGMP, whereas no effect of 8-Br-cGMP was observed for mutant S533A. Inhibition of calcium currents by 8-Br-cGMP was also observed in the additional presence of the alpha(2)delta subunit for wild type channels but not for the mutant S533A. These results indicate that cGMP causes inhibition of L-type calcium channel currents by phosphorylation of the alpha(1C) subunit at position Ser(533) via the action of protein kinase G. PMID- 10692405 TI - Transient translocation and activation of protein phosphatase 2A during mast cell secretion. AB - Okadaic acid inhibits secretion from mast cells, suggesting a regulatory role for protein Ser/Thr phosphatases type I (PP1) and/or 2A (PP2A) in the secretory process. In unstimulated RBL-2H3 cells, okadaic acid pretreatment inhibited PP2A activity in both cytosol and membrane fractions, but inhibition of secretion correlated with inhibition of membrane-bound rather than cytosolic PP2A activity. Okadaic acid had very little effect on PP1 activity. Stimulation of RBL-2H3 cells by antigen led to the activity and amount of PP2A in the membrane fraction increasing nearly 2-fold. In contrast, there was little change in the activity or distribution of PP1. Importantly, the translocation of PP2A was transient, coinciding with or marginally preceding the peak rate of secretion, suggesting a link between PP2A translocation, activity, and secretion. Phorbol 12-myristate 13 acetate plus the calcium ionophore A23187 induced a slower, prolonged rate of secretion that coincided with a similarly protracted translocation of PP2A to the membrane fraction. PP2A translocation is not the only event required for secretion as translocation was also induced by phorbol 12-myristate 13-acetate, without resulting in secretion. These results indicate that increased protein dephosphorylation in the membrane fraction mediated by PP2A is required for mast cell secretion. To our knowledge, this is the first demonstration of a signal mediated, rapid, transient translocation and activation of PP2A in membranes in any system. PMID- 10692406 TI - ARA9 modifies agonist signaling through an increase in cytosolic aryl hydrocarbon receptor. AB - The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that mediates the effects of agonists like 2,3, 7,8-tetrachlorodibenzo-p-dioxin. In the current model for AHR signaling, the unliganded receptor is found in the cytosol as part of a complex with a dimer of the 90-kDa heat shock protein and an immunophilin-like molecule, ARA9. In yeast, expression of ARA9 results in an increase in the maximal agonist response and a leftward shift in the AHR dose response curve. To better understand the mechanism by which ARA9 modifies AHR signal transduction, we performed a series of coexpression experiments in yeast and mammalian cells. Our results demonstrate that ARA9's influence on AHR signaling is not due to inhibition of a membrane pump or modification of the receptor's transactivation properties. Using receptor photoaffinity labeling experiments, we were able to show that ARA9 enhances AHR signal transduction by increasing the available AHR binding sites within the cytosolic compartment of the cell. Our evidence suggests that ARA9's effects are related to its role as a cellular chaperone; i.e. we observed that expression of ARA9 increases the fraction of AHR in the cytosol and also stabilized the receptor under heat stress. PMID- 10692407 TI - Membrane perturbation and fusion pore formation in influenza hemagglutinin mediated membrane fusion. A new model for fusion. AB - Low pH-induced fusion mediated by the hemagglutinin (HA) of influenza virus involves conformational changes in the protein that lead to the insertion of a "fusion peptide" domain of this protein into the target membrane and is thought to perturb the membrane, triggering fusion. By using whole virus, purified HA, or HA ectodomains, we found that shortly after insertion, pores of less than 26 A in diameter were formed in liposomal membranes. As measured by a novel assay, these pores stay open, or continue to close and open, for minutes to hours and persist after pH neutralization. With virus and purified HA, larger pores, allowing the leakage of dextrans, were seen at times well after insertion. For virus, dextran leakage was simultaneous with lipid mixing and the formation of "fusion pores," allowing the transfer of dextrans from the liposomal to the viral interior or vice versa. Pores did not form in the viral membrane in the absence of a target membrane. Based on these data, we propose a new model for fusion, in which HA initially forms a proteinaceous pore in the target, but not in the viral membrane, before a lipidic hemifusion intermediate is formed. PMID- 10692408 TI - HeLa cells are phenotypically limiting in cyclin E/CDK2 for efficient human papillomavirus DNA replication. AB - Human papillomaviral (HPV) origin-containing plasmids replicate efficiently in human 293 cells or cell extracts in the presence of HPV origin-recognition protein E2 and replication initiation protein E1, whereas cervical carcinoma derived, HPV-18-positive HeLa cells or cell extracts support HPV DNA replication poorly. We recently showed that HPV-11 E1 interacts with cyclin/cyclin-dependent kinase (cdk) complexes through an RXL motif and is a substrate for these kinases. E1 mutations in this motif or in candidate cdk phosphorylation sites are impaired in replication, suggesting a role for cdks in HPV replication. We now demonstrate that one limiting activity in HeLa cells is cyclin E/CDK2. Purified cyclin E/CDK2 or cyclin E/CDK3 complex, but not other cdks, partially complemented HeLa cell extracts. Cyclin E/CDK2 expression vectors also enhanced transient HPV replication in HeLa cells. HeLa cell-derived HPV-18 E1 protein is truncated at the carboxyl terminus but can associate with cyclin E/CDK2. This truncated E1 was replication-incompetent and inhibited cell-free HPV replication. These results indicate that HeLa cells are phenotypically limiting in cyclin E/CDK2 for efficient HPV replication, most likely due to sequestration by the endogenous, defective HPV-18 E1 protein. Further analyses of the regulation of HPV E1 and HPV replication by cyclin E may shed light on the roles of cyclin E/CDK2 in cellular DNA replication. PMID- 10692409 TI - Mutational analysis of basic residues in the rat vesicular acetylcholine transporter. Identification of a transmembrane ion pair and evidence that histidine is not involved in proton translocation. AB - The function of positively charged residues and the interaction of positively and negatively charged residues of the rat vesicular acetylcholine transporter (rVAChT) were studied. Changing Lys-131 in transmembrane domain helix 2 (TM2) to Ala or Leu eliminated transport activity, with no effect on vesamicol binding. However, replacement by His or Arg retained transport activity, suggesting a positive charge in this position is critical. Mutation of His-444 in TM12 or His 413 in the cytoplasmic loop between TM10 and TM11 was without effect on ACh transport, but vesamicol binding was reduced with His-413 mutants. Changing His 338 in TM8 to Ala or Lys did not effect ACh transport, however replacement with Cys or Arg abolished activity. Mutation of both of the transmembrane histidines or all three of the luminal loop histidines showed no change in acetylcholine transport. The mutant H338A/D398N between oppositely charged residues in transmembrane domains showed no vesamicol binding, however the charge reversal mutant H338D/D398H restored binding. This suggests that His-338 forms an ion pair with Asp-398. The charge neutralizing mutant K131A/D425N or the charge exchanged mutant K131D/D425K did not restore ACh transport. Taken together these results provide new insights into the tertiary structure in VAChT. PMID- 10692410 TI - RNA and DNA hydrolysis are catalyzed by the influenza virus endonuclease. AB - The influenza virus polymerase complex contains a metal ion-dependent endonuclease activity, which generates short capped RNA primer molecules from capped RNA precursors. Previous studies have provided evidence for a two-metal ion mechanism of RNA cleavage, and the data are consistent with a direct interaction of a divalent metal ion with the catalytic water molecule. To refine the model of this active site, we have generated a series of DNA, RNA, and DNA RNA chimeric molecules to study the role of the 2'-hydroxy groups on nucleic acid substrates of the endonuclease. We could observe specific cleavage of nucleic acid substrates devoid of any 2'-hydroxy groups if they contained a cap structure (m7GpppG) at the 5'-end. The capped DNA endonuclease products were functional as primers for transcription initiation by the influenza virus polymerase. The apparent cleavage rates were about 5 times lower with capped DNA substrates as compared with capped RNA substrates. Cleavage rates with DNA substrates could be increased to RNA levels by substituting the deoxyribosyl moieties immediately 5' and 3' of the cleavage site with ribosyl moieties. Similarly, cleavage rates of RNA substrates could be lowered to DNA levels by exchanging the same two ribosyl groups with deoxyribosyl groups at the cleavage site. These results demonstrate that the 2'-hydroxy groups are not essential for binding and cleavage of nucleic acids by the influenza virus endonuclease, but small differences of the nucleic acid conformation in the endonuclease active site can influence the overall rate of hydrolysis. The observed relative cleavage rates with DNA and RNA substrates argue against a direct interaction of a catalytic metal ion with a 2'-hydroxy group in the endonuclease active site. PMID- 10692411 TI - Diffusible ligand all-trans-retinal activates opsin via a palmitoylation dependent mechanism. AB - In rhodopsin's function as a photoreceptor, 11-cis-retinal is covalently bound to Lys(296) via a protonated Schiff base. 11-cis/all-trans photoisomerization and relaxation through intermediates lead to the metarhodopsin II photoproduct, which couples to transducin (G(t)). Here we have analyzed a different signaling state that arises from noncovalent binding of all-trans-retinal (atr) to the aporeceptor opsin and enhances the very low opsin activity by several orders of magnitude. Like with metarhodopsin II, coupling of G(t) to opsin-atr is sensitive to competition by synthetic peptides from the COOH termini of both G(t)alpha and G(t)gamma. However, atr does not compete with 11-cis-retinal incorporation into the Lys(296) binding site and formation of the light-sensitive pigment. Blue light illumination fails to photorevert opsin-atr to the ground state. Thus noncovalently bound atr has no access to the light-dependent binding site and reaction pathway. Moreover, in contrast to light-dependent signaling, removal of the palmitoyl anchors at Cys(322) and Cys(323) in the rhodopsin COOH terminus impairs the atr-stimulated activity. Repalmitoylation by autoacylation with palmitoyl-coenzyme A restores most of the original activity. We hypothesize that the palmitoyl moieties are part of a second binding pocket for the chromophore, mediating hydrophobic interactions that can activate a large part of the catalytic receptor/G-protein interface. PMID- 10692412 TI - cDNA cloning, purification, and characterization of mouse liver selenocysteine lyase. Candidate for selenium delivery protein in selenoprotein synthesis. AB - Selenocysteine lyase (SCL) (EC 4.4.1.16) is a pyridoxal 5'-phosphate-dependent enzyme that specifically catalyzes the decomposition of L-selenocysteine to L alanine and elemental selenium. The enzyme was proposed to function as a selenium delivery protein to selenophosphate synthetase in selenoprotein biosynthesis (Lacourciere, G. M., and Stadtman, T. C. (1998) J. Biol. Chem. 273, 30921-30926). We purified SCL from pig liver and determined its partial amino acid sequences. Mouse cDNA clones encoding peptides resembling pig SCL were found in the expressed sequence tag data base, and their sequences were used as probes to isolate full-length mouse liver cDNA. The cDNA for mouse SCL (mSCL) was determined to be 2,172 base pairs in length, containing an open reading frame encoding a polypeptide chain of 432 amino acid residues (M(r) 47, 201). We also determined the sequence of the N-terminal region of putative human SCL. These enzymes were shown to be distantly related in primary structure to NifS, which catalyzes the desulfurization of L-cysteine to provide sulfur for iron-sulfur clusters. The recombinant mSCL overproduced in Escherichia coli was a homodimer with the subunit M(r) of 47,000. The enzyme was pyridoxal phosphate-dependent and highly specific to L-selenocysteine (the k(cat)/K(m) value for L-selenocysteine was about 4,200 times higher than that for L-cysteine). Reverse transcriptase polymerase chain reaction and Western blot analyses revealed that mSCL is cytosolic and predominantly exists in the liver, kidney, and testis, where mouse selenophosphate synthetase is also abundant, supporting the view that mSCL functions in cooperation with selenophosphate synthetase in selenoprotein synthesis. This is the first report of the primary structure of mammalian SCL. PMID- 10692413 TI - Biosynthetic studies of the glycopeptide teicoplanin by (1)H and (13)C NMR. AB - The biosynthesis of the glycopeptide antibiotic teicoplanin was studied by growing a teicoplanin producing strain of Actinoplanes teichomyceticus (ATCC 31121) on glucose containing either 34.0% [1-(13)C]glucose or 9.7% [U (13)C]glucose. The fractional enrichment pattern of teicoplanin produced in the medium containing [1-(13)C]glucose was obtained from a one-dimensional (13)C spectrum. The enrichment pattern showed characteristic peaks indicating that amino acids 3 and 7 are derived from acetate, whereas amino acids 1, 2, 4, 5, and 6 are derived from tyrosine. Multiplet structures in heteronuclear single quantum coherence spectra of teicoplanin produced in the medium containing [U (13)C]glucose showed characteristic coupling patterns supporting these results. Fractional enrichment patterns and multiplet structures of the three sugars in teicoplanin showed that about 50% of the sugars have the same labeling pattern as the glucose substrate whereas the rest have a labeling pattern showing that they are reassembled, probably from precursors in the primary metabolism. PMID- 10692414 TI - Distant downstream sequence determinants can control N-tail translocation during protein insertion into the endoplasmic reticulum membrane. AB - We have studied the membrane insertion of ProW, an Escherichia coli inner membrane protein with seven transmembrane segments and a large periplasmic N terminal tail, into endoplasmic reticulum (ER)-derived dog pancreas microsomes. Strikingly, significant levels of N-tail translocation is seen only when a minimum of four of the transmembrane segments are present; for constructs with fewer transmembrane segments, the N-tail remains mostly nontranslocated and the majority of the molecules adopt an "inverted" topology where normally nontranslocated parts are translocated and vice versa. N-tail translocation can also be promoted by shortening of the N-tail and by the addition of positively charged residues immediately downstream of the first trasnmembrane segment. We conclude that as many as four consecutive transmembrane segments may be collectively involved in determining membrane protein topology in the ER and that the effects of downstream sequence determinants may vary depending on the size and charge of the N-tail. We also provide evidence to suggest that the ProW N tail is translocated across the ER membrane in a C-to-N-terminal direction. PMID- 10692415 TI - Hydrolysis of nucleoside triphosphates other than ATP by nitrogenase. AB - The hydrolysis of ATP to ADP and P(i) is an integral part of all substrate reduction reactions catalyzed by nitrogenase. In this work, evidence is presented that nitrogenases isolated from Azotobacter vinelandii and Clostridium pasteurianum can hydrolyze MgGTP, MgITP, and MgUTP to their respective nucleoside diphosphates at rates comparable to those measured for MgATP hydrolysis. The reactions were dependent on the presence of both the iron (Fe) protein and the molybdenum-iron (MoFe) protein. The oxidation state of nitrogenase was found to greatly influence the nucleotide hydrolysis rates. MgATP hydrolysis rates were 20 times higher under dithionite reducing conditions (approximately 4,000 nmol of MgADP formed per min/mg of Fe protein) as compared with indigo disulfonate oxidizing conditions (200 nmol of MgADP formed per min/mg of Fe protein). In contrast, MgGTP, MgITP, and MgUTP hydrolysis rates were significantly higher under oxidizing conditions (1,400-2,000 nmol of MgNDP formed per min/mg of Fe protein) as compared with reducing conditions (80-230 nmol of MgNDP formed per min/mg of Fe protein). The K(m) values for MgATP, MgGTP, MgUTP, and MgITP hydrolysis were found to be similar (330-540 microM) for both the reduced and oxidized states of nitrogenase. Incubation of Fe and MoFe proteins with each of the MgNTP molecules and AlF(4)(-) resulted in the formation of non-dissociating protein-protein complexes, presumably with trapped AlF(4)(-) x MgNDP. The implications of these results in understanding how nucleotide hydrolysis is coupled to substrate reduction in nitrogenase are discussed. PMID- 10692416 TI - Effects of interferon-gamma and lipopolysaccharide on macrophage iron metabolism are mediated by nitric oxide-induced degradation of iron regulatory protein 2. AB - Iron regulatory proteins (IRP-1 and IRP-2) control the synthesis of transferrin receptors (TfR) and ferritin by binding to iron-responsive elements, which are located in the 3'-untranslated region and the 5'-untranslated region of their respective mRNAs. Cellular iron levels affect binding of IRPs to iron-responsive elements and consequently expression of TfR and ferritin. Moreover, NO(*), a redox species of nitric oxide that interacts primarily with iron, can activate IRP-1 RNA binding activity resulting in an increase in TfR mRNA levels. Recently we found that treatment of RAW 264.7 cells (a murine macrophage cell line) with NO(+) (nitrosonium ion, which causes S-nitrosylation of thiol groups) resulted in a rapid decrease in RNA binding of IRP-2 followed by IRP-2 degradation, and these changes were associated with a decrease in TfR mRNA levels (Kim, S., and Ponka, P. (1999) J. Biol. Chem. 274, 33035-33042). In this study, we demonstrated that stimulation of RAW 264.7 cells with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) increased IRP-1 binding activity, whereas RNA binding of IRP-2 decreased and was followed by a degradation of this protein. Moreover, the decrease of IRP-2 binding/protein levels was associated with a decrease in TfR mRNA levels in LPS/IFN-gamma-treated cells, and these changes were prevented by inhibitors of inducible nitric oxide synthase. Furthermore, LPS/IFN-gamma stimulated RAW 264.7 cells showed increased rates of ferritin synthesis. These results suggest that NO(+)-mediated degradation of IRP-2 plays a major role in iron metabolism during inflammation. PMID- 10692417 TI - Reconstitution of the Ste24p-dependent N-terminal proteolytic step in yeast a factor biogenesis. AB - The yeast mating pheromone a-factor precursor contains an N-terminal extension and a C-terminal CAAX motif within which multiple posttranslational processing events occur. A recently discovered component in a-factor processing is Ste24p/Afc1p, a multispanning endoplasmic reticulum membrane protein that contains an HEXXH metalloprotease motif. Our in vivo genetic characterization of this protein has demonstrated roles for Ste24p in both the N-terminal and C terminal proteolytic processing of the a-factor precursor. Here, we present evidence that the N-terminal proteolysis of the a-factor precursor P1 can be accurately reconstituted in vitro using yeast membranes. We show that this activity is dependent on Ste24p and is abolished by mutation of the Ste24p HEXXH metalloprotease motif or by mutation of the a-factor P1 substrate at a residue adjacent to the N-terminal P1 cleavage site. We also demonstrate that N-terminal proteolysis of the P1 a-factor precursor requires Zn(2+) as a co-factor and can be inhibited by the addition of the metalloprotease inhibitor 1,10 orthophenanthroline. Our results are consistent with Ste24p itself being the P1- >P2 a-factor protease or a limiting activator of this activity. Interestingly, we also show that the human Ste24 homolog expressed in yeast can efficiently promote the N-terminal processing of a-factor in vivo and in vitro, thus establishing a factor as a surrogate substrate in the absence of known human substrates. The results reported here, together with the previously reported in vitro reconstitution of Ste24p-dependent CAAX processing, provide a system for examining the potential bifunctional roles of yeast Ste24p and its homologs. PMID- 10692418 TI - Substitution of leucine 28 with histidine in the Escherichia coli transcription factor FNR results in increased stability of the [4Fe-4S](2+) cluster to oxygen. AB - To understand the role of the [4Fe-4S](2+) cluster in controlling the activity of the Escherichia coli transcription factor FNR (fumarate nitrate reduction) during changes in O(2) availability, we have characterized a mutant FNR protein containing a substitution of Leu-28 with His (FNR-L28H) which, unlike its wild type (WT) counterpart, is functional under aerobic growth conditions. The His-28 substitution appears to stabilize the [4Fe-4S](2+) cluster of FNR-L28H in the presence of O(2) because air-exposed FNR-L28H did not undergo the rapid [4Fe 4S](2+) to [2Fe-2S](2+) cluster conversion or concomitant loss in site-specific DNA binding and dimerization, which are characteristic of WT-FNR under these conditions. This increased cluster stability was not a result of His-28 replacing the WT-FNR cluster ligands because substitution of any of these four Cys residues (cysteine 20, 23, 29, or 122) with Ser resulted in [4Fe-4S](2+) cluster-deficient preparations of FNR-L28H. The Mossbauer spectra of FNR-L28H indicated that the coordination environment of the [4Fe-4S](2+) cluster did not differ from that of WT-FNR. Whole cell Mossbauer spectroscopy showed that aerobically grown cells overexpressing FNR-L28H had levels of the FNR species containing the [4Fe-4S](2+) cluster similar to those of cells grown under anaerobic conditions. Thus, the increase in cluster stability is sufficient to allow accumulation of the [4Fe 4S](2+) cluster form of FNR-L28H under aerobic conditions and provides a reasonable explanation for why this mutant protein is functional under aerobic growth conditions. From these results, we present a model to explain how WT-FNR is normally inactivated under aerobic growth conditions. PMID- 10692419 TI - Identification and characterization of a novel cAMP receptor protein in the cyanobacterium Synechocystis sp. PCC 6803. AB - Three open reading frames of Synechocystis sp. PCC 6803 encoding a domain homologous with the cAMP binding domain of bacterial cAMP receptor protein were analyzed. These three open reading frames, sll1371, sll1924, and slr0593, which were named sycrp1, sycrp2, and sypk, respectively, were expressed in Escherichia coli as His-tagged or glutathione S-transferase fusion proteins and purified, and their biochemical properties were investigated. The results obtained for equilibrium dialysis measurements using these recombinant proteins suggest that SYCRP1 and SYPK show a binding affinity for cAMP while SYCRP2 does not. The dissociation constant of His-tagged SYCRP1 for cAMP is approximately 3 microM. A cross-linking experiment using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide revealed that His-tagged SYCRP1 forms a homodimer, and the presence or absence of cAMP does not affect the formation of the homodimer. The amino acid sequence reveals that SYCRP1 has a domain similar to the DNA binding domain of bacterial cAMP receptor protein in the COOH-terminal region. Consistent with this, His tagged SYCRP1 forms a complex with DNA that contains the consensus sequence for E. coli cAMP receptor protein in the presence of cAMP. These results strongly suggest that SYCRP1 is a novel cAMP receptor protein. PMID- 10692420 TI - Retroviral transfection of Madin-Darby canine kidney cells with human MDR1 results in a major increase in globotriaosylceramide and 10(5)- to 10(6)-fold increased cell sensitivity to verocytotoxin. Role of p-glycoprotein in glycolipid synthesis. AB - Retroviral infection of the Madin-Darby canine kidney (MDCK) renal cell line with human MDR1 cDNA, encoding the P-glycoprotein (P-gp) multidrug resistance efflux pump, induces a major accumulation of the glycosphingolipid (GSL), globotriaosylceramide (Galalpha1-4Galbeta1-4glucosylceramide-Gb(3)), the receptor for the E. coli-derived verotoxin (VT), to effect a approximately million-fold increase in cell sensitivity to VT. The shorter chain fatty acid isoforms of Gb(3) (primarily C16 and C18) are elevated and VT is internalized to the endoplasmic reticulum/nuclear envelope as we have reported for other hypersensitive cell lines. P-gp (but not MRP) inhibitors, e.g. ketoconazole or cyclosporin A (CsA) prevented the increased Gb(3) and VT sensitivity, concomitant with increased vinblastine sensitivity. Gb(3) synthase was not significantly elevated in MDR1-MDCK cells and was not affected by CsA. In MDR1-MDCK cells, synthesis of fluorescent N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-aminocaproyl (NBD)-lactosylceramide (LacCer) and NBD-Gb(3) via NBD-glucosylceramide (GlcCer) from exogenous NBD-C(6)-ceramide, was prevented by CsA. We therefore propose that P-gp can mediate GlcCer translocation across the bilayer, from the cytosolic face of the Golgi to the lumen, to provide increased substrate for the lumenal synthesis of LacCer and subsequently Gb(3). These results provide a molecular mechanism for the observed increased sensitivity of multidrug-resistant tumors to VT and emphasize the potential of verotoxin as an antineoplastic. Two strains (I and II) of MDCK cells, which differ in their glycolipid profile, have been described. The original MDR1-MDCK parental cell was not specified, but the MDR1 MDCK GSL phenotype and glycolipid synthase activities indicate MDCK-I cells. However, the partial drug resistance of MDCK-I cells precludes their being the parental cell. We speculate that the retroviral transfection per se, or the subsequent selection for drug resistance, selected a subpopulation of MDCK-I cells in the parental MDCK-II cell culture and that drug resistance in MDR1-MDCK cells is thus a result of both MDR1 expression and a second, previously unrecognized, component, likely the high level of GlcCer synthesis in these cells. PMID- 10692421 TI - Functional heterogeneity of small ubiquitin-related protein modifiers SUMO-1 versus SUMO-2/3. AB - Post-translational modification marked by the covalent attachment of the ubiquitin-like protein SUMO-1/SMT3C has been implicated in a wide variety of cellular processes. Recently, two cDNAs encoding proteins related to SUMO-1 have been identified in human and mouse. The functions and regulation of these proteins, known as SUMO-2/SMT3A and SUMO-3/SMT3B, remain largely uncharacterized. We describe herein quantitative and qualitative distinctions between SUMO-1 and SUMO-2/3 in vertebrate cells. Much of this was accomplished through the application of an antibody that recognizes SUMO-2 and -3, but not SUMO-1. This antibody detected multiple SUMO-2/3-modified proteins and revealed that, together, SUMO-2 and -3 constitute a greater percentage of total cellular protein modification than does SUMO-1. Intriguingly, we found that there was a large pool of free, non-conjugated SUMO-2/3 and that the conjugation of SUMO-2/3 to high molecular mass proteins was induced when the cells were subjected to protein damaging stimuli such as acute temperature fluctuation. In addition, we demonstrated that SUMO-2/3 conjugated poorly, if at all, to a major SUMO-1 substrate, the Ran GTPase-activating protein RanGAP1. Together, these results support the concept of important distinctions between the SUMO-2/3 and SUMO-1 conjugation pathways and suggest a role for SUMO-2/3 in the cellular responses to environmental stress. PMID- 10692422 TI - Transcriptional activation of the cyclooxygenase-2 gene in endotoxin-treated RAW 264.7 macrophages. AB - Cyclooxygenase-2 (COX-2), the enzyme primarily responsible for induced prostaglandin synthesis, is an immediate early gene induced by endotoxin in macrophages. We investigated the cis-acting elements of the COX-2 5'-flanking sequence, the transcription factors and signaling pathways responsible for transcriptional activation of the COX-2 gene in endotoxin-treated murine RAW 264.7 macrophages. Luciferase reporter constructs with alterations in presumptive cis-acting transcriptional regulatory elements demonstrate that the cyclic AMP response element and two nuclear factor interleukin-6 (CCAAT/enhancer-binding protein (C/EBP)) sites of the COX-2 promoter are required for optimal endotoxin dependent induction. In contrast, the E-box and NF-kappaB sites are not required for endotoxin-dependent induction. Inhibition of endotoxin-induced NF-kappaB activation by expression of an inhibitor-kappaB alpha mutant does not block endotoxin-dependent COX-2 reporter activity. Overexpression of c-Jun, C/EBPbeta, and C/EBPdelta enhances induction of the COX-2 reporter, while overexpression of cyclic AMP-response element-binding protein or "dominant negative" C/EBPbeta represses COX-2 induction. In addition, endotoxin rapidly and transiently elicits c-Jun phosphorylation in RAW 264.7 macrophages. Cotransfection of the COX-2 reporter with dominant negative expression vectors shows that endotoxin-induced COX-2 gene expression requires signaling through a Ras-independent pathway involving the adapter protein ECSIT and the signaling kinases MEKK1 and JNK. In contrast, endotoxin-induced COX-2 reporter activity is not blocked by overexpression of dominant-negative forms of Raf-1, ERK1, or ERK2. PMID- 10692423 TI - The catalytic subunit of phosphoinositide 3-kinase: requirements for oncogenicity. AB - The retroviral oncogene p3k (v-p3k) of avian sarcoma virus 16 (ASV16) codes for the catalytic subunit of phosphoinositide (PI) 3-kinase, p110alpha. The v-P3k protein is oncogenic in vivo and in vitro; its cellular counterpart, c-P3k, lacks oncogenicity. Fusion of viral Gag sequences to the amino terminus of c-P3k activates the transforming potential. Activation can also be achieved by the addition of a myristylation signal to the amino terminus or of a farnesylation signal to the carboxyl terminus of c-P3k. A mutated myristylation signal was equally effective; it also caused a strong increase in the kinase activity of P3k. Mutations that inactivate lipid kinase activity abolish oncogenicity. The transforming activity of P3k is correlated with the ability to induce activating phosphorylation in Akt. Point mutations and amino-terminal deletions recorded in v-P3k were shown to be irrelevant to the activation of oncogenic potential. Interactions of P3k with the regulatory subunit of PI 3-kinase, p85, or with Ras are not required for transformation. These results support the conclusion that the oncogenicity of P3k depends on constitutive lipid kinase activity. Akt is an important and probably essential downstream component of the oncogenic signal from P3k. PMID- 10692424 TI - Alkyl-dihydroxyacetonephosphate synthase. Presence and role of flavin adenine dinucleotide. AB - Alkyl-dihydroxyacetonephosphate synthase is a peroxisomal enzyme involved in ether lipid synthesis. It catalyzes the exchange of the acyl chain in acyl dihydroxyacetonephosphate for a long chain fatty alcohol, yielding the first ether linked intermediate, i.e. alkyl-dihydroxyacetonephosphate, in the pathway of ether lipid biosynthesis. Although this reaction is not a net redox reaction, the amino acid sequence of the enzyme suggested the presence of a flavin adenine dinucleotide (FAD)-binding domain. In this study we show that alkyl dihydroxyacetonephosphate synthase contains an essential FAD molecule as cofactor, which is evidenced by fluorescence properties, UV-visible absorption spectra and the observation that the enzyme activity is dependent on the presence of this cofactor in a coupled in vitro transcription/translation assay. Furthermore, we could demonstrate that the FAD cofactor directly participates in catalysis. Upon incubation of the enzyme with the substrate palmitoyl dihydroxyacetonephosphate, the flavin moiety is reduced, indicating that in this initial step the substrate is oxidized. Stopped flow experiments show that the reduction of the flavin moiety is a monophasic process yielding a oxygen stable, reduced enzyme species. Upon addition of hexadecanol to the reduced enzyme species, the flavin moiety is efficiently reoxidized. A hypothetical reaction mechanism is proposed that is consistent with the data in this paper and with previous studies. PMID- 10692425 TI - Molecular determinants of pH sensitivity of the type IIa Na/P(i) cotransporter. AB - Type II Na/P(i) cotransporters play key roles in epithelial P(i) transport and thereby contribute to overall P(i) homeostasis. Renal proximal tubular brush border membrane expresses the IIa isoform, whereas the IIb isoform is preferentially expressed in small intestinal brush border membrane of mammals. IIa and IIb proteins are predicted to contain eight transmembrane domains with the N- and C-terminal tails facing the cytoplasm. They differ in their pH dependences: the activity of IIa increases at higher pH, whereas the IIb shows no or a slightly opposite pH dependence. To determine the structural domains responsible for the difference in pH sensitivity, mouse IIa and IIb chimeras were constructed, and their pH dependence was characterized. A region between the fourth and fifth transmembrane domains was required for conferring pH sensitivity to the IIa-mediated Na/P(i) cotransport. Sequence comparison (IIa versus IIb) of the third extracellular loops revealed a stretch of three charged amino acids in IIa (REK) replaced by uncharged residues in IIb (GNT). Introduction of the uncharged GNT sequence (by REK) in IIa abolished its pH dependence, whereas introduction of the charged REK stretch in IIb (by GNT) led to a pH dependence similar to IIa. These findings suggest that charged residues within the third extracellular loop are involved in the pH sensitivity of IIa Na/P(i) cotransporter. PMID- 10692426 TI - Selenoprotein P expression, purification, and immunochemical characterization. AB - Most selenoproteins contain a single selenocysteine residue per polypeptide chain, encoded by an in-frame UGA codon. Selenoprotein P is unique in that its mRNA encodes 10-12 selenocysteine residues, depending on species. In addition to the high number of selenocysteines, the protein is cysteine- and histidine-rich. The function of selenoprotein P has remained elusive, in part due to the inability to express the recombinant protein. This has been attributed to presumed inefficient translation through the selenocysteine/stop codons. Herein, we report for the first time the expression of recombinant rat selenoprotein P in a transiently transfected human epithelial kidney cell line, as well as the endogenously expressed protein from HepG2 and Chinese hamster ovary cells. The majority of the expressed protein migrates with the predicted 57-kDa size of full length glycosylated selenoprotein P. Based on the histidine-rich nature of selenoprotein P, we have purified the recombinant and endogenously expressed proteins using nickel-agarose affinity chromatography. We show that the recombinant rat and endogenous human proteins react in Western blotting and immunoprecipitation assays with commercial anti-histidine antibodies. The ability to express, purify, and immunochemically detect the recombinant protein provides a foundation for investigating the functions and efficiency of expression of this intriguing protein. PMID- 10692427 TI - Functional heterodimerization of prolactin and growth hormone receptors by ovine placental lactogen. AB - Although homo- or heterodimerization are common mechanisms for activation of cytokine receptors, cross-talk between two distinct receptors in this superfamily has been never shown. Here we show a physiologically relevant example indicating that such an interaction does occurs, thus raising the hypothesis that heterodimerization between distinct cytokine receptors may be a novel mechanism contributing to the diversity of cytokine signaling. These findings were documented using both surface plasmon resonance and gel filtration experiments and show that ovine placental lactogen (PL) heterodimerizes the extracellular domains (ECDs) of ruminant growth hormone receptor (GHR) and prolactin receptor (PRLR). We also show that PL or PL analogues that exhibit little or no activity in cells transfected with PRLRs and no activity in cells transfected with ovine GHRs exhibit largely enhanced activity in cells cotransfected with both PRLRs and GHRs. Furthermore, chimeric receptors consisting of cytosolic and transmembrane part of ovine GHR or ovine PRLR and ECDs of human granulocyte-macrophage colony stimulating factor receptor (GM-CSFR) alpha or beta were constructed. Upon transfection into Chinese hamster ovary cells along with reporter luciferase gene and stimulation by GM-CSF, a significant increase in luciferase activity occurred when GM-CSFR-alpha-PRLR and GM-CSFR-beta-GHR or GM-CSFR-alpha-GHR and GM-CSRR beta-PRLR were cotransfected. In conclusion, we show that ovine PL is capable of functional heterodimerization of GHR and PRLR and that when their cytosolic parts, coupled to the ECD of GM-CSF receptors, are heterodimerized by GM-CSF, they are capable of transducing biological signal. PMID- 10692428 TI - Kinetic and pharmacological properties of human brain Na(+)/H(+) exchanger isoform 5 stably expressed in Chinese hamster ovary cells. AB - The recently cloned Na(+)/H(+) exchanger isoform 5 (NHE5) is expressed predominantly in brain, yet little is known about its functional properties. To facilitate its characterization, a full-length cDNA encoding human NHE5 was stably transfected into NHE-deficient Chinese hamster ovary AP-1 cells. Pharmacological analyses revealed that H(+)(i)-activated (22)Na(+) influx mediated by NHE5 was inhibited by several classes of drugs (amiloride compounds, 3-methylsulfonyl-4-piperidinobenzoyl guanidine methanesulfonate, cimetidine, and harmaline) at half-maximal concentrations that were intermediate to those determined for the high affinity NHE1 and the low affinity NHE3 isoforms, but closer to the latter. Kinetic analyses showed that the extracellular Na(+) dependence of NHE5 activity followed a simple hyperbolic relationship with an apparent affinity constant (K(Na)) of 18.6 +/- 1.6 mM. By contrast to other NHE isoforms, NHE5 also exhibited a first-order dependence on the intracellular H(+) concentration, achieving half-maximal activation at pH 6.43 +/- 0.08. Extracellular monovalent cations, such as H(+) and Li(+), but not K(+), acted as effective competitive inhibitors of (22)Na(+) influx by NHE5. In addition, the transport activity of NHE5 was highly dependent on cellular ATP levels. Overall, these functional features distinguish NHE5 from other family members and closely resemble those of an amiloride-resistant NHE isoform identified in hippocampal neurons. PMID- 10692429 TI - Dynamics of protein-tyrosine phosphatases in rat adipocytes. AB - Protein-tyrosine phosphatases (PTPases) play a key role in maintaining the steady state tyrosine phosphorylation of the insulin receptor (IR) and its substrate proteins such as insulin receptor substrate 1 (IRS-1). However, the PTPase(s) that inactivate IR and IRS-1 under physiological conditions remain unidentified. Here, we analyze the subcellular distribution in rat adipocytes of several PTPases thought to be involved in the counterregulation of insulin signaling. We found that the transmembrane enzymes, protein-tyrosine phosphatase (PTP)-alpha and leukocyte common antigen-related (LAR), were detected predominantly in the plasma membrane and to a lesser extent in the heavy microsomes, a distribution similar to that of insulin receptor. PTP-1B and IRS-1 were present in light microsomes and cytosol, whereas SHPTP2/Syp was exclusively cytosolic. Insulin induced a redistribution of PTP-alpha from the plasma membrane to the heavy microsomes in a parallel fashion with the receptor. The distribution of PTP-1B in the light microsomes from resting adipocytes was similar to that of IRS-1 as determined by sucrose velocity gradient fractionation. Analysis of the catalytic activity of partially purified rat adipocyte PTP-alpha and LAR and recombinant PTP-1B showed that all three PTPases dephosphorylate IR. When a mix of IR/IRS-1 was used as a substrate, PTP-1B was particularly effective in dephosphorylating IRS-1. Considering that IR and IRS-1 can be dephosphorylated in internal membrane compartments from rat adipocytes (Kublaoui, B., Lee, J., and Pilch, P.F. (1995) J. Biol. Chem. 270, 59-65) and that PTP-alpha and PTP-1B are the respective PTPases in these fractions, we conclude that these PTPases are responsible for the counterregulation of insulin signaling there, whereas both LAR and PTP-alpha may act upon cell surface insulin receptors. PMID- 10692430 TI - Reconstitution of virus-mediated expression of interferon alpha genes in human fibroblast cells by ectopic interferon regulatory factor-7. AB - Type I interferons constitute an important part of the innate immune response against viral infection. Unlike the expression of interferon (IFN) B gene, the expression of IFNA genes is restricted to the lymphoid cells. Both IFN regulatory factor 3 and 7 (IRF-3 and IRF-7) were suggested to play positive roles in these genes expression. However, their role in the differential expression of individual subtypes of human IFNA genes is unknown. Using various IFNA reporter constructs in transient transfection assay we found that overexpression of IRF-3 in virus infected 2FTGH cells selectively activated IFNA1 VRE, whereas IRF-7 was able to activate IFNA1, A2, and A4. The binding of recombinant IRF-7 and IRF-3 to these VREs correlated with their transcriptional activation. Nuclear proteins from infected and uninfected IRF-7 expressing 2FTGH cells formed multiple DNA protein complexes with IFNA1 VRE, in which two unique DNA-protein complexes containing IRF-7 were detected. In 2FTGH cells, virus stimulated expression of IFNB gene but none of the IFNA genes. Reconstitution of IRF-7 synthesis in these cells resulted, upon virus infection, in the activation of seven endogenous IFNA genes in which IFNA1 predominated. These studies suggest that IRF-7 is a critical determinant for the induction of IFNA genes in infected cells. PMID- 10692431 TI - The organization of aggrecan in human articular cartilage. Evidence for age related changes in the rate of aggregation of newly synthesized molecules. AB - The effect of age on the incorporation of newly synthesized aggrecan into the extracellular matrix of human articular cartilage was investigated. This property was measured in a pulse-chase explant culture system by determining the distribution of radiolabeled molecules ([(35)S]sulfate-labeled) between a nondissociating extract (phosphate-buffered saline), which extracts mainly nonaggregated macromolecules, and a dissociating extract (4 M GnHCl) containing mainly aggrecan that was complexed in situ with hyaluronan. The rate of incorporation of aggrecan into aggregates was much slower in mature cartilage than in tissue obtained from younger individuals. Furthermore, autoradiography showed that in mature cartilage, newly synthesized aggrecan is not transported from the pericellular environment within the first 18 h of chase culture, whereas in immature cartilage, it moves into the intercellular space during the same period, i.e. aggrecan is processed in the extracellular space very differently in young and adult articular cartilage. Experiments were also performed to show that the interaction of link protein with newly synthesized aggrecan depends on the maturity of the G(1) domain of aggrecan. This investigation has shown that the extracellular aggregation of aggrecan in adult human articular cartilage involves a number of intermediate structures. These have not been identified in the very young cartilage obtained from laboratory animals or in porcine and bovine articular cartilage obtained from the abattoir. PMID- 10692432 TI - The C terminus of SNAP25 is essential for Ca(2+)-dependent binding of synaptotagmin to SNARE complexes. AB - The plasma membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins syntaxin and synaptosome-associated protein of 25 kDa (SNAP25) and the vesicle SNARE protein vesicle-associated membrane protein (VAMP) are essential for a late Ca(2+)-dependent step in regulated exocytosis, but their precise roles and regulation by Ca(2+) are poorly understood. Botulinum neurotoxin (BoNT) E, a protease that cleaves SNAP25 at Arg(180)-Ile(181), completely inhibits this late step in PC12 cell membranes, whereas BoNT A, which cleaves SNAP25 at Gln(197)-Arg(198), is only partially inhibitory. The difference in toxin effectiveness was found to result from a reversal of BoNT A but not BoNT E inhibition by elevated Ca(2+) concentrations. BoNT A treatment essentially increased the Ca(2+) concentration required to activate exocytosis, which suggested a role for the C terminus of SNAP25 in the Ca(2+) regulation of exocytosis. Synaptotagmin, a proposed Ca(2+) sensor for exocytosis, was found to bind SNAP25 in a Ca(2+)-stimulated manner. Ca(2+)-dependent binding was abolished by BoNT E treatment, whereas BoNT A treatment increased the Ca(2+) concentration required for binding. The C terminus of SNAP25 was also essential for Ca(2+) dependent synaptotagmin binding to SNAP25. syntaxin and SNAP25.syntaxin.VAMP SNARE complexes. These results clarify classical observations on the Ca(2+) reversal of BoNT A inhibition of neurosecretion, and they suggest that an essential role for the C terminus of SNAP25 in regulated exocytosis is to mediate Ca(2+)-dependent interactions between synaptotagmin and SNARE protein complexes. PMID- 10692433 TI - Cdk2-dependent and -independent pathways in E2F-mediated S phase induction. AB - The transcription factor E2F plays an important role in G(1) to S phase transition in the higher eukaryotic cell cycle. Although a number of E2F inducible genes have been identified, the biochemical cascades from E2F to the S phase entry remain to be investigated. In this study, we generated stably transfected mouse NIH3T3 cells that express exogenous human E2F-1 under the control of a heavy metal-inducible metallothionein promoter and analyzed the molecular mechanism of the E2F-1-mediated initiation of chromosomal DNA replication. Ectopic E2F-1 expression in cells arrested in G(0)/G(1) by serum deprivation enabled them to progress through G(1) and to enter S phase. During the G(1) progression, mouse cyclin E, but little of cyclin D1, was induced to express, which subsequently activated Cdk2. Experiments using the Cdk inhibitory proteins p27, p18, and p19 proved that the activity of Cdk2, but not of Cdk4, was required for S phase entry mediated by E2F-1. Minichromosome maintenance proteins (MCM) 4 and 7, the components of the DNA-replication initiation complex (RC), were constitutively expressed during the cell cycle, although the MCM genes are well known E2F-inducible genes. However, tight association of these two proteins with chromatin depended upon ectopic E2F-1 expression. In contrast, the Cdc45 protein, another RC component, which turned out to be a transcriptional target of E2Fs, was induced to express and subsequently bound to chromatin in response to E2F-1. Experiments utilizing a chemical Cdk-specific inhibitor, butyrolactone I, revealed that Cdk2 activity was required only for chromatin binding of the Cdc45 proteins, and not for the expression of Cdc45 or chromatin binding of MCM4 and 7. These results indicate that at least two separate pathways function downstream of E2F to initiate S phase; one depends upon the activity of Cdk2 and the other does not. PMID- 10692434 TI - Dityrosine formation outcompetes tyrosine nitration at low steady-state concentrations of peroxynitrite. Implications for tyrosine modification by nitric oxide/superoxide in vivo. AB - Formation of peroxynitrite from NO and O-(*2) is considered an important trigger for cellular tyrosine nitration under pathophysiological conditions. However, this view has been questioned by a recent report indicating that NO and O-(*2) generated simultaneously from (Z)-1-(N-[3-aminopropyl]-N-[4-(3 aminopropylammonio)butyl]-amino) diazen-1-ium-1,2-diolate] (SPER/NO) and hypoxanthine/xanthine oxidase, respectively, exhibit much lower nitrating efficiency than authentic peroxynitrite (Pfeiffer, S. and Mayer, B. (1998) J. Biol. Chem. 273, 27280-27285). The present study extends those earlier findings to several alternative NO/O-(*2)-generating systems and provides evidence that the apparent lack of tyrosine nitration by NO/O-(*2) is due to a pronounced decrease of nitration efficiency at low steady-state concentrations of authentic peroxynitrite. The decrease in the yields of 3-nitrotyrosine was accompanied by an increase in the recovery of dityrosine, showing that dimerization of tyrosine radicals outcompetes the nitration reaction at low peroxynitrite concentrations. The observed inverse dependence on peroxynitrite concentration of dityrosine formation and tyrosine nitration is predicted by a kinetic model assuming that radical formation by peroxynitrous acid homolysis results in the generation of tyrosyl radicals that either dimerize to yield dityrosine or combine with (*)NO(2) radical to form 3-nitrotyrosine. The present results demonstrate that very high fluxes (>2 microM/s) of NO/O-(*2) are required to render peroxynitrite an efficient trigger of tyrosine nitration and that dityrosine is a major product of tyrosine modification caused by low steady-state concentrations of peroxynitrite. PMID- 10692435 TI - DNA methyltransferase inhibition induces the transcription of the tumor suppressor p21(WAF1/CIP1/sdi1). AB - Previous lines of evidence have shown that inhibition of DNA methyltransferase (MeTase) can arrest tumor cell growth; however, the mechanisms involved were not clear. In this manuscript we show that out of 16 known tumor suppressors and cell cycle regulators, the cyclin-dependent kinase inhibitor p21 is the only tumor suppressor induced in the human lung cancer cell line, A549, following inhibition of DNA MeTase by a novel DNA MeTase antagonist or antisense oligonucleotides. The rapid induction of p21 expression points to a mechanism that does not involve demethylation of p21 promoter. Consistent with this hypothesis, we show that part of the CpG island upstream of the endogenous p21 gene is unmethylated and that the expression of unmethylated p21 promoter luciferase reporter constructs is induced following inhibition of DNA MeTase. These results are consistent with the hypothesis that the level of DNA MeTase in a cell can control the expression of a nodal tumor suppressor by a mechanism that does not involve DNA methylation. PMID- 10692436 TI - Ca(2+)-dependent and Ca(2+)-independent calmodulin binding sites in erythrocyte protein 4.1. Implications for regulation of protein 4.1 interactions with transmembrane proteins. AB - In vitro protein binding assays identified two distinct calmodulin (CaM) binding sites within the NH(2)-terminal 30-kDa domain of erythrocyte protein 4.1 (4.1R): a Ca(2+)-independent binding site (A(264)KKLWKVCVEHHTFFRL) and a Ca(2+)-dependent binding site (A(181)KKLSMYGVDLHKAKDL). Synthetic peptides corresponding to these sequences bound CaM in vitro; conversely, deletion of these peptides from a 30 kDa construct reduced binding to CaM. Thus, 4.1R is a unique CaM-binding protein in that it has distinct Ca(2+)-dependent and Ca(2+)-independent high affinity CaM binding sites. CaM bound to 4.1R at a stoichiometry of 1:1 both in the presence and absence of Ca(2+), implying that one CaM molecule binds to two distinct sites in the same molecule of 4.1R. Interactions of 4.1R with membrane proteins such as band 3 is regulated by Ca(2+) and CaM. While the intrinsic affinity of the 30-kDa domain for the cytoplasmic tail of erythrocyte membrane band 3 was not altered by elimination of one or both CaM binding sites, the ability of Ca(2+)/CaM to down regulate 4. 1R-band 3 interaction was abrogated by such deletions. Thus, regulation of protein 4.1 binding to membrane proteins by Ca(2+) and CaM requires binding of CaM to both Ca(2+)-independent and Ca(2+)-dependent sites in protein 4.1. PMID- 10692437 TI - NBP-45, a novel nucleosomal binding protein with a tissue-specific and developmentally regulated expression. AB - Here we characterize a novel murine nuclear protein, which we named NBP-45, that is related to the ubiquitous nuclear proteins HMG-14/-17, binds specifically to nucleosome core particles, and can function as a transcriptional activator. NBP 45 mRNA is expressed at low levels and in variable amounts in all mouse tissues tested but is especially abundant in RNA extracted from 7-day-old mouse embryos, suggesting that it functions in early embryonic development. NBP-45 is composed of 406 amino acids and is encoded by a single size transcript. The region spanning the N-terminal 85 amino acids contains three segments that are highly homologous to functionally important domains in the HMG-14/-17 protein family: the nuclear localization signal, the nucleosome binding domain, and the chromatin unfolding domain. The protein region spanning the C-terminal 321 amino acids has a 42% content of negatively charged residues. The first 23 amino acids contain a region necessary for nuclear entry of the protein, the region spanning residues 12-40 is the main nucleosomal binding domain of the protein, and the negatively charged, C-terminal domain is necessary for transcription activation. The functional domains of NBP-45 are indicative of a nuclear protein that binds to nucleosomes, thereby creating a chromatin region of high local negative charge. Our studies establish the nucleosomal binding domain as a protein motif that is present in other than just the ubiquitous HMG-14/-17 proteins. We suggest that the nucleosomal binding domain motif is a protein module that facilitates binding to nucleosomes in chromatin. PMID- 10692438 TI - Importance of the hinge region between alpha-helix F and the main part of serpins, based upon identification of the epitope of plasminogen activator inhibitor type 1 neutralizing antibodies. AB - The serpin plasminogen activator inhibitor type 1 (PAI-1) is an important protein in the regulation of fibrinolysis and inhibits its target proteinases through formation of a covalent complex. In the present study, we have identified the epitope of two PAI-1 neutralizing monoclonal antibodies (MA-33H1F7 and MA 55F4C12). Based upon differential cross-reactivity data of these monoclonals with PAI-1 from different species and on a sequence alignment between these PAI-1s, combined with the three-dimensional structure, we predicted that the residues Glu(128)-Val(129)-Glu(130)-Arg(131) and Lys(154) (at the hinge region between alpha-helix F and the main part of the PAI-1-molecule) might form the major site of interaction. Therefore a variety of alanine mutants were generated and evaluated for their affinity toward both monoclonal antibodies. The affinity constants of MA-55F4C12 and MA-33H1F7 for PAI-1 were 2.7 +/- 1.6 x 10(9) M(-1) and 5.4 +/- 1.7 x 10(9) M(-1), respectively, but decreased between 13- and 270 fold upon mutation of Lys(154) to Ala(154) or Glu(128)-Val(129)-Glu(130)-Arg(131) to Ala-Ala-Ala-Ala. The combined mutations (PAI-1-EVER/K), however, resulted in an absence of binding to either of the antibodies. Both antibodies bound to PAI-1 wt/t-PA complexes with a similar affinity as to PAI-1-wt (K(A) = 4-5 x 10(9) M( 1)). The epitope localization reveals the molecular basis for the neutralizing properties of both monoclonal antibodies. In addition, it provides new insights into the validity of various models that have been proposed for the serpin/proteinase complex, excluding full insertion of the reactive-site loop. PMID- 10692439 TI - Cardiovascular basic helix loop helix factor 1, a novel transcriptional repressor expressed preferentially in the developing and adult cardiovascular system. AB - We have cloned a cardiovascular-restricted basic helix-loop-helix factor that interacts with arylhydrocarbon receptor nuclear translocator (ARNT) in a yeast two-hybrid screen. Cardiovascular helix-loop-helix factor 1 (CHF1) is distantly related to the hairy family of transcriptional repressors. We analyzed its expression pattern during mouse embryo development. At day 8.5, the expression of CHF1 is first detected in the primitive ventricle of the primordial heart tube and persists throughout gestation. In rat hearts, this expression is down regulated after birth, concurrent with terminal differentiation of cardiomyocytes. In the developing vasculature, CHF1 first appears in the dorsal aorta at day 9.0, which precedes the reported expression of smooth muscle cell markers, and persists into adulthood. In an in vitro system of smooth muscle cell differentiation, CHF1 mRNA was barely detectable in undifferentiated cells but was induced highly in differentiated smooth muscle cells. To determine whether CHF1 might affect the function of ARNT, we performed transfection studies. Co transfection of CHF1 inhibited ARNT/EPAS1-dependent transcription by 85%, and this inhibition is dose-dependent. In electrophoretic mobility studies, CHF1 inhibited the binding of the ARNT/EPAS1 heterodimer to its target site. Our data suggest that CHF1 functions as a transcriptional repressor and may play an important role in cardiovascular development. PMID- 10692440 TI - G protein modulation of N-type calcium channels is facilitated by physical interactions between syntaxin 1A and Gbetagamma. AB - The direct modulation of N-type calcium channels by G protein betagamma subunits is considered a key factor in the regulation of neurotransmission. Some of the molecular determinants that govern the binding interaction of N-type channels and Gbetagamma have recently been identified (see, i.e., Zamponi, G. W., Bourinet, E., Nelson, D., Nargeot, J., and Snutch, T. P. (1997) Nature 385, 442-446); however, little is known about cellular mechanisms that modulate this interaction. Here we report that a protein of the presynaptic vesicle release complex, syntaxin 1A, mediates a crucial role in the tonic inhibition of N-type channels by Gbetagamma. When syntaxin 1A was coexpressed with (N-type) alpha(1B) + alpha(2)-delta + beta(1b) channels in tsA-201 cells, the channels underwent a 18 mV negative shift in half-inactivation potential, as well as a pronounced tonic G protein inhibition as assessed by its reversal by strong membrane depolarizations. This tonic inhibition was dramatically attenuated following incubation with botulinum toxin C, indicating that syntaxin 1A expression was indeed responsible for the enhanced G protein modulation. However, when G protein betagamma subunits were concomitantly coexpressed, the toxin became ineffective in removing G protein inhibition, suggesting that syntaxin 1A optimizes, rather than being required for G protein modulation of N-type channels. We also demonstrate that Gbetagamma physically binds to syntaxin 1A, and that syntaxin 1A can simultaneously interact with Gbetagamma and the synprint motif of the N-type channel II-III linker. Taken together, our experiments suggest a mechanism by which syntaxin 1A mediates a colocalization of G protein betagamma subunits and N type calcium channels, thus resulting in more effective G protein coupling to, and regulation of, the channel. Thus, the interactions between syntaxin, G proteins, and N-type calcium channels are part of the structural specialization of the presynaptic terminal. PMID- 10692441 TI - An isoform of kalirin, a brain-specific GDP/GTP exchange factor, is enriched in the postsynaptic density fraction. AB - Communication between membranes and the actin cytoskeleton is an important aspect of neuronal function. Regulators of actin cytoskeletal dynamics include the Rho like small GTP-binding proteins and their exchange factors. Kalirin is a brain specific protein, first identified through its interaction with peptidylglycine alpha-amidating monooxygenase. In this study, we cloned rat Kalirin-7, a 7 kilobase mRNA form of Kalirin. Kalirin-7 contains nine spectrin-like repeats, a Dbl homology domain, and a pleckstrin homology domain. We found that the majority of Kalirin-7 protein is associated with synaptosomal membranes, but a fraction is cytosolic. We also detected higher molecular weight Kalirin proteins. In rat cerebral cortex, Kalirin-7 is highly enriched in the postsynaptic density fraction. In primary cultures of neurons, Kalirin-7 is detected in spine-like structures, while other forms of Kalirin are visualized in the cell soma and throughout the neurites. Kalirin-7 and its Dbl homology-pleckstrin homology domain induce formation of lamellipodia and membrane ruffling, when transiently expressed in fibroblasts, indicative of Rac1 activation. Using Rac1, the Dbl homology-pleckstrin homology domain catalyzed the in vitro exchange of bound GDP with GTP. Kalirin-7 is the first guanine-nucleotide exchange factor identified in the postsynaptic density, where it is positioned optimally to regulate signal transduction pathways connecting membrane proteins and the actin cytoskeleton. PMID- 10692442 TI - Chat, a Cas/HEF1-associated adaptor protein that integrates multiple signaling pathways. AB - Cas (Crk-associated substrate) and HEF1 (human enhancer of filamentation) are related adaptor proteins that function in integrin-mediated cell adhesion and antigen receptor signaling pathways. We report here a molecular cloning of Chat (Cas/HEF1-associated signal transducer) that associates with Cas and HEF1. Chat is a 78-kDa signaling molecule with an N-terminal SH2 domain and is expressed in a wide range of tissues. In hematopoietic cells, a 115-kDa isoform of Chat (Chat H) was specifically expressed. Chat is associated with Cas in brain, and Chat-H is associated with HEF1 in splenocytes. Deletion analyses revealed that Chat and Cas are associated with each other by their C-terminal domains. Treatment of PC12 cells with epidermal growth factor or nerve growth factor increased the phosphorylation level of Chat. This increase was suppressed by an inhibitor of mitogen-activated protein (MAP) kinase kinase, PD98059, suggesting the phosphorylation of Chat by MAP kinase. In Chat-overexpressed COS7 cells, the activity of c-Jun N-terminal kinase was up-regulated. After the epidermal growth factor stimulation, Chat and Cas were colocalized with actin filaments at ruffling membranes. These findings suggest that Chat transduces signals of tyrosine kinases and MAP kinase to Cas signaling pathway. PMID- 10692443 TI - Differential roles of the Src homology 2 domains of phospholipase C-gamma1 (PLC gamma1) in platelet-derived growth factor-induced activation of PLC-gamma1 in intact cells. AB - Upon stimulation of cells with platelet-derived growth factor (PDGF), phospholipase C-gamma1 (PLC-gamma1) binds to the tyrosine-phosphorylated PDGF receptor through one or both of its Src homology 2 (SH2) domains, is phosphorylated by the receptor kinase, and is thereby activated to hydrolyze phosphatidylinositol 4, 5-bisphosphate. Association of PLC-gamma1 with the insoluble subcellular fraction is also enhanced in PDGF-stimulated cells. The individual roles of the two SH2 domains of PLC-gamma1 in mediating the interaction between the enzyme and the PDGF receptor have now been investigated by functionally disabling each domain. A critical Arg residue in each SH2 domain was mutated to Ala. Both wild-type and mutant PLC-gamma1 proteins were transiently expressed in a PLC-gamma1-deficient fibroblast cell line, and these transfected cells were stimulated with PDGF. The mutant protein in which the COOH terminal SH2 domain was disabled bound to the PDGF receptor. Accordingly, it was phosphorylated by the receptor, catalyzed the production of inositol phosphates, and mobilized intracellular calcium to extents similar to (but slightly less than) those observed with the wild-type enzyme. In contrast, the mutant in which the NH(2)-terminal SH2 domain was impaired did not bind to the PDGF receptor and consequently was neither phosphorylated nor activated. These results suggest that the NH(2)-terminal SH2 domain, but not the COOH-terminal SH2 domain, of PLC gamma1 is required for PDGF-induced activation of PLC-gamma1. Functional impairment of the SH2 domains did not affect the PDGF-induced redistribution of PLC-gamma1, suggesting that recruitment of PLC-gamma1 to the particulate fraction does not involve the SH2 domains. PMID- 10692444 TI - Calcineurin is downstream of the inositol 1,4,5-trisphosphate receptor in the apoptotic and cell growth pathways. AB - The inositol 1,4,5-trisphosphate receptor (IP(3)R) is a calcium (Ca(2+)) release channel found on the endoplasmic reticulum of virtually all types of cells. Human T lymphocytes (Jurkat) that are made deficient in IP(3)R do not generate Ca(2+) signals in response to T cell receptor stimulation, fail to translocate the nuclear factor for activated T cells to the nucleus, and are remarkably resistant to induction of apoptosis with CD95 (Fas), dexamethasone, gamma irradiation, and T cell receptor stimulation using anti-CD3 antibody. Expression of constitutively active calcineurin A in IP(3)R-deficient T cells restored nuclear factor for activated T cells translocation to the nucleus and dephosphorylation of Bad and rendered the cells sensitive to apoptotic inducers. Induction of apoptosis required both active calcineurin A (DeltaCnA) and activation-dependent colocalization of CnA with its substrate. Thus, the Ca(2+)-dependent phosphatase calcineurin (CnA) is downstream of the IP(3)R in both the cell growth and apoptotic signaling pathways. PMID- 10692445 TI - NFkappaB mediates apoptosis through transcriptional activation of Fas (CD95) in adenoviral hepatitis. AB - NFkappaB is an essential survival factor in several physiological conditions such as embryonal liver development and liver regeneration. However, NFkappaB is also a main mediator of the cellular response to a variety of extracellular stress stimuli, and it has been shown that some viral-induced host cell apoptosis appears to be dependent on NFkappaB activation. The activation of NFkappaB upon viral infection may be a rapid way of initiating an innate immune response against the viral particles. We have assessed the role of NFkB during the early phase of adenoviral hepatitis in a nude mouse model using an adenoviral vector expressing a mutant form of IkappaBalpha. Administration of a LacZ-expressing adenoviral vector induces NFkB DNA and correlates with the up-regulation of Fas (CD95) mRNA, but not FasL (CD95L) mRNA, during the early phase of adenoviral hepatitis. The rapid increase in NFkappaB DNA binding after adenoviral infection of the liver could be very effectively inhibited by IkappaBalpha. Compared with the LacZ control virus, the IkappaBalpha-expressing adenoviral vector inhibits the increase of Fas (CD95) mRNA expression, in particular in the very early phase of the hepatitis. Reporter gene experiments in hepatoma cell lines with a Fas promoter-luciferase construct indicated that the repression of Fas (CD95) mRNA by IkappaBalpha was transcriptionally mediated. The functional relevance of the NFkappaB-dependent increase in Fas (CD95) transcription was assessed by caspase 3 assays and terminal dUTP nick-end labeling tests. Compared with the control, IkappaBalpha adenoviral infection resulted in reduced caspase 3 activity during the early phase of viral hepatitis and in a prevention of liver cell apoptosis 24 h after adenoviral administration. Therefore our study demonstrates a new pro apoptotic function of NFkappaB in Fas (CD95)-mediated apoptosis of hepatocytes. Interestingly, NFkappaB mediates liver cell apoptosis upon viral infection even in a phase where tumor necrosis factor-alpha is already induced, as shown by the time curves of tumor necrosis factor-alpha serum levels. Therefore, the pro- or anti-apoptotic role of NFkappaB appears to be more determined by the nature of the death stimulus than by the origin of the tissue. PMID- 10692446 TI - A novel phosphatidylinositol-phospholipase C of Trypanosoma cruzi that is lipid modified and activated during trypomastigote to amastigote differentiation. AB - The phosphoinositide (PI)-specific phospholipase C gene (TcPI-PLC) of the protozoan parasite Trypanosoma cruzi was cloned, sequenced, expressed in Escherichia coli, and the protein product (TcPI-PLC) was shown to have enzymatic characteristics similar to those of mammalian delta-type PI-PLCs. The TcPI-PLC gene is expressed at high levels in the epimastigote and amastigote stages of the parasite, and its expression is induced during the differentiation of trypomastigotes into amastigotes, where TcPI-PLC associates with the plasma membrane and increases its catalytic activity. In contrast to other PI-PLCs described so far, the deduced amino acid sequence of TcPI-PLC revealed some unique features such as an N-myristoylation consensus sequence at its amino terminal end, lack of an apparent pleckstrin homology domain and a highly charged linker region between the catalytic X and Y domains. TcPI-PLC is lipid modified in vivo, as demonstrated by metabolic labeling with [(3)H]myristate and [(3)H]palmitate and fatty acid analysis of the immunoprecipitated protein, and may constitute the first example of a new group of PI-PLCs. PMID- 10692447 TI - Cholesterol oxidation switches the internalization pathway of endothelin receptor type A from caveolae to clathrin-coated pits in Chinese hamster ovary cells. AB - We investigated the mechanism of endothelin receptor type A (ETA) internalization in Chinese hamster ovary cells using two assays; flow cytometric quantification of cell surface myc-ETA and in situ localization of Cy5-labeled ET-1. In both assays, agonist-dependent internalization of myc-ETA was inhibited by nystatin and filipin, both of which disrupt internalization via caveolae, whereas it was barely affected by chlorpromazine and hypertonic sucrose, both of which disrupt internalization via clathrin-coated pits. In addition to myc-ETA, ET-1 caused intracellular translocation of caveolin-1 and this translocation was also blocked by nystatin but not by chlorpromazine. These results strongly argue that ETA is internalized via caveolae but not clathrin-coated pits. Treatment of the cells with cholesterol oxidase reduced cellular cholesterol and caused intracellular translocation of caveolin-1 but did not affect cell surface localization of myc ETA. In cholesterol oxidase-treated cells, however, both chlorpromazine and hypertonic sucrose effectively blocked ET-1-induced myc-ETA internalization and nystatin was less effective than in untreated cells. Accordingly, expression of a dominant negative form of beta-arrestin blocked myc-ETA internalization in cholesterol oxidase-treated cells but not in untreated cells. These results suggest that, in Chinese hamster ovary cells, 1) agonist-occupied ETA can be internalized either via caveolae or clathrin-coated pits; 2) of the two, the former is the default pathway; and 3) the oxidative state of cell surface cholesterol is one of the factors involved in the pathway selection. PMID- 10692448 TI - The human 20-kDa 5'-(CGG)(n)-3'-binding protein is targeted to the nucleus and affects the activity of the FMR1 promoter. AB - Previous reports have described the human DNA CGG repeat-binding protein (CGGBP1 or p20), which binds specifically to nonmethylated, but not to methylated, 5' (CGG)(n)-3' repeats in the promoter of the fragile X mental retardation 1 (FMR1) gene. The results of transfection experiments into human HeLa cells using a p20 green fluorescent protein fusion construct indicate that the p20 protein is targeted to the nucleus. By deletion analyses, a nuclear localization signal has been found between amino acids 80 and 84. Deletions between amino acids 69 and 71 and between 95 and 167 interfere with 5'-(CGG)(n)-3' binding. The results of electrophoretic mobility shift assays using DNA with 5'-(CGG)(n)-3' repeats of different lengths render it likely that oligomers of the p20 protein bind to the repeat. In cotransfection experiments, the activity of the FMR1 promoter is reduced by the presence of p20. Upon transfection of the p20 cDNA construct into HeLa cells, transcription of the endogenous FMR1 gene is decreased. The green fluorescent protein-p20 fusion protein associates preferentially with the telomeres of the short arms of human chromosomes 13, 14, 15, 21, and 22. Their telomeres carry the genes for the 28 S rRNA, which contain 5'-(CGG)(n)-3' repeats. The translated region of the p20 gene from three healthy, five fragile X syndrome, and five premutation-carrying individuals has been sequenced, but mutations have not been detected. PMID- 10692449 TI - Cloning and functional characterization of novel large conductance calcium activated potassium channel beta subunits, hKCNMB3 and hKCNMB4. AB - We present the cloning and characterization of two novel calcium-activated potassium channel beta subunits, hKCNMB3 and hKCNMB4, that are enriched in the testis and brain, respectively. We compare and contrast the steady state and kinetic properties of these beta subunits with the previously cloned mouse beta1 (mKCNMB1) and the human beta2 subunit (hKCNMB2). Once inactivation is removed, we find that hKCNMB2 has properties similar to mKCNMB1. hKCNMB2 slows Hslo1 channel gating and shifts the current-voltage relationship to more negative potentials. hKCNMB3 and hKCNMB4 have distinct effects on slo currents not observed with mKCNMB1 and hKCNMB2. Although we found that hKCNMB3 does interact with Hslo channels, its effects on Hslo1 channel properties were slight, increasing Hslo1 activation rates. In contrast, hKCNMB4 slows Hslo1 gating kinetics, and modulates the apparent calcium sensitivity of Hslo1. We found that the different effects of the beta subunits on some Hslo1 channel properties are calcium-dependent. mKCNMB1 and hKCNMB2 slow activation at 1 microM but not at 10 microM free calcium concentrations. hKCNMB4 decreases Hslo1 channel openings at low calcium concentrations but increases channel openings at high calcium concentrations. These results suggest that beta subunits in diverse tissue types fine-tune slo channel properties to the needs of a particular cell. PMID- 10692450 TI - Thrombin inhibits tumor cell growth in association with up-regulation of p21(waf/cip1) and caspases via a p53-independent, STAT-1-dependent pathway. AB - Thrombin, a multifunctional protein, has been found to be involved in cellular mitogenesis, tumor growth, and metastasis, in addition to its well known effects on the initiation of platelet aggregation and secretion and the conversion of fibrinogen to fibrin to form blood clots. These properties of thrombin rely on its action as a serine protease, which cleaves the N-terminal region of a 7 transmembrane G protein receptor (protease-activated receptor, PAR-1), thus exposing a tethered end hexapeptide sequence capable of activating its receptor. Little is known about its effect on genes that regulate the cell cycle. This study was undertaken to investigate the possible mechanisms by which thrombin regulates tumor cell growth in several tumor cell lines: human CHRF megakaryocyte, DU145 prostate, MDAMB231 and MCF7 breast, U3A fibrosarcoma, and 2 murine fibroblast cell lines, MEFp53(-/-) and CD STAT(-/-). We have found that thrombin under the conditions of culture employed inhibits cell growth by both up regulation of p21(waf/cip1) and induction of caspases via its PAR-1 receptor. The increased expression of p21(waf/cip1) by thrombin was p53 independent, STAT1 dependent, and protein synthesis independent. This was associated with tyrosine phosphorylation of JAK2 and STAT1, and nuclear translocation of STAT1. Induction of apoptosis is also PAR-1-specific, STAT1-dependent, and associated with up regulation of caspases 1, 2, and 3. Our study establishes, for the first time, a link between PAR-1 receptor activation with the STAT signal pathway, which leads to cell cycle control and apoptosis. This observation broadens our understanding of the mechanism of PAR-1 activation and its effect on cell growth, and could possibly lead to therapeutic approaches for the treatment of cancer. PMID- 10692451 TI - Determination of the disulfide bond arrangement of Newcastle disease virus hemagglutinin neuraminidase. Correlation with a beta-sheet propeller structural fold predicted for paramyxoviridae attachment proteins. AB - Disulfide bonds stabilize the structure and functions of the hemagglutinin neuraminidase attachment glycoprotein (HN) of Newcastle disease virus. Until this study, the disulfide linkages of this HN and structurally similar attachment proteins of other members of the paramyxoviridae family were undefined. To define these linkages, disulfide-linked peptides were produced by peptic digestion of purified HN ectodomains of the Queensland strain of Newcastle disease virus, isolated by reverse phase high performance liquid chromatography, and analyzed by mass spectrometry. Analysis of peptides containing a single disulfide bond revealed Cys(531)-Cys(542) and Cys(172)-Cys(196) linkages and that HN ectodomains dimerize via Cys(123). Another peptide, with a chain containing Cys(186) linked to a chain containing Cys(238), Cys(247), and Cys(251), was cleaved at Met(249) with cyanogen bromide. Subsequent tandem mass spectrometry established Cys(186) Cys(247) and Cys(238)-Cys(251) linkages. A glycopeptide with a chain containing Cys(344) linked to a chain containing Cys(455), Cys(461), and Cys(465) was treated sequentially with peptide-N-glycosidase F and trypsin. Further treatment of this peptide by one round of manual Edman degradation or tandem mass spectrometry established Cys(344)-Cys(461) and Cys(455)-Cys(465) linkages. These data, establishing the disulfide linkages of all thirteen cysteines of this protein, are consistent with published predictions that the paramyxoviridae HN forms a beta-propeller structural fold. PMID- 10692452 TI - Epsin binds to clathrin by associating directly with the clathrin-terminal domain. Evidence for cooperative binding through two discrete sites. AB - Epsin is a recently identified protein that appears to play an important role in clathrin-mediated endocytosis. The central region of epsin 1, the so-called DPW domain, binds to the heterotetrameric AP-2 adaptor complex by associating directly with the globular appendage of the alpha subunit. We have found that this central portion of epsin 1 also associates with clathrin. The interaction with clathrin is direct and not mediated by epsin-bound AP-2. Alanine scanning mutagenesis shows that clathrin binding depends on the sequence (257)LMDLADV located within the epsin 1 DPW domain. This sequence, related to the known clathrin-binding sequences in the adaptor beta subunits, amphiphysin, and beta arrestin, facilitates the association of epsin 1 with the terminal domain of the clathrin heavy chain. Unexpectedly, inhibiting the binding of AP-2 to the GST epsin DPW fusion protein by progressively deleting DPW triplets but leaving the LMDLADV sequence intact, diminishes the association of clathrin in parallel with AP-2. Because the beta subunit of the AP-2 complex also contains a clathrin binding site, optimal association with soluble clathrin appears to depend on the presence of at least two distinct clathrin-binding sites, and we show that a second clathrin-binding sequence (480)LVDLD, located within the carboxyl-terminal segment of epsin 1, also interacts with clathrin directly. The LMDLADV and LVDLD sequences act cooperatively in clathrin recruitment assays, suggesting that they bind to different sites on the clathrin-terminal domain. The evolutionary conservation of similar clathrin-binding sequences in several metazoan epsin-like molecules suggests that the ability to establish multiple protein-protein contacts within a developing clathrin-coated bud is an important aspect of epsin function. PMID- 10692453 TI - The role of the conserved box E residues in the active site of the Escherichia coli type I signal peptidase. AB - Type I signal peptidases are integral membrane proteins that function to remove signal peptides from secreted and membrane proteins. These enzymes carry out catalysis using a serine/lysine dyad instead of the prototypical serine/histidine/aspartic acid triad found in most serine proteases. Site directed scanning mutagenesis was used to obtain a qualitative assessment of which residues in the fifth conserved region, Box E, of the Escherichia coli signal peptidase I are critical for maintaining a functional enzyme. First, we find that there is no requirement for activity for a salt bridge between the invariant Asp-273 and the Arg-146 residues. In addition, we show that the conserved Ser-278 is required for optimal activity as well as conserved salt bridge partners Asp-280 and Arg-282. Finally, Gly-272 is essential for signal peptidase I activity, consistent with it being located within van der Waals proximity to Ser-278 and general base Lys-145 side-chain atoms. We propose that replacement of the hydrogen side chain of Gly-272 with a methyl group results in steric crowding, perturbation of the active site conformation, and specifically, disruption of the Ser-90/Lys-145 hydrogen bond. A refined model is proposed for the catalytic dyad mechanism of signal peptidase I in which the general base Lys 145 is positioned by Ser-278, which in turn is held in place by Asp-280. PMID- 10692454 TI - A-Myb up-regulates Bcl-2 through a Cdx binding site in t(14;18) lymphoma cells. AB - In follicular lymphoma, bcl-2 is translocated to the immunoglobulin heavy chain locus leading to deregulation of bcl-2 expression. We examined the role of Myb proteins in the regulation of bcl-2 expression in lymphoma cells. We showed that A-Myb up-regulates bcl-2 promoter activity. Northern and Western analyses demonstrated that A-Myb was expressed in the DHL-4 t(14;18) cell line. In t(14;18) cells and mature B cells, A-Myb up-regulated bcl-2 expression, whereas B and c-Myb had little effect on bcl-2 gene expression. Deletion analysis of the bcl-2 5'-region identified a region responsive to A-Myb in t(14;18) cells. A potential binding site for the Cdx homeodomain proteins was located in this sequence. Analysis of the A-Myb-responsive region by UV cross-linking experiments revealed that a 32-kDa protein formed a complex with this region, but direct binding by Myb proteins could not be demonstrated. A-Myb could be recovered along with Cdx2 when nuclear extracts were passed over the Cdx site. Mutagenesis of the Cdx binding site abolished binding by the 32-kDa protein and significantly reduced the ability of A-Myb to induce bcl-2 expression. A strong induction of bcl-2 P2 promoter activity was observed in cotransfection studies of DHL-4 cells with the A-Myb and Cdx2 expression vectors, and increased endogenous Bcl-2 protein expression was observed in B cells transfected with A-Myb and/or Cdx2 expression constructs. PMID- 10692455 TI - Phosphatidylglycerol is involved in the dimerization of photosystem II. AB - Photosystem II core dimers (450 kDa) and monomers (230 kDa) consisting of CP47, CP43, the D1 and D2 proteins, the extrinsic 33-kDa subunit, and the low molecular weight polypeptides PsbE, PsbF, PsbH, PsbI, PsbK, PsbL, PsbTc, and PsbW were isolated by sucrose density gradient centrifugation. The photosystem II core dimers were treated with phospholipase A2 (PL-A2), which cuts phosphatidylglycerol (PG) and phosphatidylcholine molecules at the sn-2 position. The PL-A2-treated dimers dissociated into two core monomers and further, yielding a CP47-D1-D2 subcomplex and CP43. Thin layer chromatography showed that photosystem II dimers contained four times more PG than their monomeric counterparts but with similar levels of phosphatidylcholine. Consistent with this was the finding that, compared with monomers, the dimers contained a higher level of trans-hexadecanoic fatty acid (C16:1Delta3tr), which is specific to PG of the thylakoid membrane. Moreover, treatment of dimers with PL-A2 increased the free level of this fatty acid specific to PG compared with untreated dimers. Further evidence that PG is involved in stabilizing the dimeric state of photosystem II comes from reconstitution experiments. Using size exclusion chromatography, it was shown that PG containing C16:1Delta3tr, but not other lipid classes, induced significant dimerization of isolated photosystem II monomers. Moreover, this dimerization was observed by electron crystallography when monomers were reconstituted into thylakoid lipids containing PG. The unit cell parameters, p2 symmetry axis, and projection map of the reconstituted dimer was similar to that observed for two-dimensional crystals of the native dimer. PMID- 10692456 TI - Tissue specificity of E subunit isoforms of plant vacuolar H(+)-ATPase and existence of isotype enzymes. AB - Immunoblot analyses and partial amino acid sequencings revealed that both the 40- (E1) and 37-kDa (E2) subunits of V-ATPase in the pea epicotyl were E subunit isoforms. Similarly, both the 35- (D1) and 29-kDa (D2) subunits were D subunit isoforms, although the similarity of the amino acid sequences is still unknown. In immunoblot analyses, two or three E subunit isoforms with molecular masses ranging from 29 to 40 kDa were detected in other plants. Two isotypes of V-ATPase from the pea epicotyl were separated by ion exchange chromatography and had subunit compositions differing only in the ratio of E1 and E2. There was a difference in the V(max) and K(m) of ATP hydrolysis between the two isotypes. E1 was scarcely detected in crude membrane fractions from the leaf and cotyledon, while E2 was detected in fractions from all of the tissues examined. The compositions of D subunit isoforms in the leaf and epicotyl were different, and the vacuolar membrane in the leaf did not contain D2. The efficiency of H(+) pumping activity in the vacuolar membrane of the leaf was higher than that of the epicotyl. The results suggest that the presence of the isoforms of D and E subunits is characteristic to plants and that the isoforms are closely related to the enzymatic properties. PMID- 10692457 TI - Syntaxin 7 mediates endocytic trafficking to late endosomes. AB - The lysosome functions are ensured by accurate membrane trafficking in the cell. We found that mouse syntaxin 7 could complement yeast vam3 and pep12 mutants defective in docking/fusion to vacuolar and prevacuolar membranes, respectively. Immunohistochemical studies showed that syntaxin 7 is localized to late endosomes, but not to early endosomes. Induced expression of mutant syntaxin 7 blocked endocytic transport from early to late endosomes but did not block the transport of cathepsin D and lamp-2 from the trans-Golgi network to lysosomes. Thus, syntaxin 7 mediates the endocytic trafficking from early endosomes to late endosomes and lysosomes. These results also suggest that the biosynthetic pathway utilizes a different machinery from that of the endocytic pathway in the docking/fusion to late endosomes. PMID- 10692458 TI - The 8-nucleotide-long RNA:DNA hybrid is a primary stability determinant of the RNA polymerase II elongation complex. AB - The sliding clamp model of transcription processivity, based on extensive studies of Escherichia coli RNA polymerase, suggests that formation of a stable elongation complex requires two distinct nucleic acid components: an 8-9-nt transcript-template hybrid, and a DNA duplex immediately downstream from the hybrid. Here, we address the minimal composition of the processive elongation complex in the eukaryotes by developing a method for promoter-independent assembly of functional elongation complex of S. cerevisiae RNA polymerase II from synthetic DNA and RNA oligonucleotides. We show that only one of the nucleic acid components, the 8-nt RNA:DNA hybrid, is necessary for the formation of a stable elongation complex with RNA polymerase II. The double-strand DNA upstream and downstream of the hybrid does not affect stability of the elongation complex. This finding reveals a significant difference in processivity determinants of RNA polymerase II and E. coli RNA polymerase. In addition, using the imperfect RNA:DNA hybrid disturbed by the mismatches in the RNA, we show that nontemplate DNA strand may reduce the elongation complex stability via the reduction of the RNA:DNA hybrid length. The structure of a "minimal stable" elongation complex suggests a key role of the RNA:DNA hybrid in RNA polymerase II processivity. PMID- 10692459 TI - 5-hydroxyconiferyl aldehyde modulates enzymatic methylation for syringyl monolignol formation, a new view of monolignol biosynthesis in angiosperms. AB - S-Adenosyl-L-methionine-dependent caffeate O-methyltransferase (COMT, EC 2.1.1.6) has traditionally been thought to catalyze the methylation of caffeate and 5- hydroxyferulate for the biosynthesis of syringyl monolignol, a lignin constituent of angiosperm wood that enables efficient lignin degradation for cellulose production. However, recent recognition that coniferyl aldehyde prevents 5 hydroxyferulate biosynthesis in lignifying tissue, and that the hydroxylated form of coniferyl aldehyde, 5-hydroxyconiferyl aldehyde, is an alternative COMT substrate, demands a re-evaluation of the role of COMT during monolignol biosynthesis. Based on recombinant aspen (Populus tremuloides) COMT enzyme kinetics coupled with mass spectrometry analysis, this study establishes for the first time that COMT is in fact a 5-hydroxyconiferyl aldehyde O-methyltransferase (AldOMT), and that 5-hydroxyconiferyl aldehyde is both the preferred AldOMT substrate and an inhibitor of caffeate and 5-hydroxyferulate methylation, as measured by K(m) and K(i) values. 5-Hydroxyconiferyl aldehyde also inhibited the caffeate and 5-hydroxyferulate methylation activities of xylem proteins from various angiosperm tree species. The evidence that syringyl monolignol biosynthesis is independent of caffeate and 5-hydroxyferulate methylation supports our previous discovery that coniferyl aldehyde prevents ferulate 5 hydroxylation and at the same time ensures a coniferyl aldehyde 5-hydroxylase (CAld5H)-mediated biosynthesis of 5-hydroxyconiferyl aldehyde. Together, our results provide conclusive evidence for the presence of a CAld5H/AldOMT-catalyzed coniferyl aldehyde 5-hydroxylation/methylation pathway that directs syringyl monolignol biosynthesis in angiosperms. PMID- 10692460 TI - Identification and characterization of Elongin A2, a new member of the Elongin family of transcription elongation factors, specifically expressed in the testis. AB - The Elongin complex stimulates the rate of transcription elongation by RNA polymerase II by suppressing the transient pausing of the polymerase at many sites along the DNA template. Elongin is composed of a transcriptionally active A subunit and two small regulatory B and C subunits, the latter of which bind stably to each other to form a binary complex that interacts with Elongin A and strongly induces its transcriptional activity. To further understand the roles of Elongin in transcriptional regulation, we attempted to identify Elongin-related proteins. Here, we report on the cloning, expression, and characterization of human Elongin A2, a novel transcription elongation factor that exhibited 47% identity and 61% similarity to Elongin A. Biochemical studies have shown that Elongin A2 stimulates the rate of transcription elongation by RNA polymerase II and is capable of forming a stable complex with Elongin BC. However, in contrast to Elongin A, its transcriptional activity is not activated by Elongin BC. Northern blot analysis revealed that Elongin A2 mRNA was specifically expressed in the testis, suggesting that Elongin A2 may regulate the transcription of testis-specific genes. PMID- 10692461 TI - Norepinephrine- and phorbol ester-induced phosphorylation of alpha(1a)-adrenergic receptors. Functional aspects. AB - Maximal adrenergic responses in Rat-1 fibroblasts expressing alpha(1a)-adrenergic receptors are not blocked by activation of protein kinase C. In contrast, activation of protein kinase C induces the phosphorylation of alpha(1b) adrenoreceptors and blocks their actions. The effect of norepinephrine and phorbol esters on alpha(1a)-adrenoreceptor phosphorylation and coupling to G proteins were studied. Both stimuli lead to dose-dependent receptor phosphorylation. Interestingly, protein kinase C activation affected to a much lesser extent the actions of alpha(1a)-adrenergic receptors than those of the alpha(1b) subtype (norepinephrine elicited increases in calcium in whole cells and [(35)S]GTPgammaS binding to membranes). Basal phosphorylation of alpha(1a) adrenergic receptors was much less than that observed with the alpha(1b) subtype. The carboxyl terminus seems to be the main domain for receptor phosphorylation. Therefore, chimeric receptors, where the carboxyl-terminal tails of alpha(1a) and alpha(1b) adrenergic receptors were exchanged, were constructed and expressed. alpha(1a)-Adrenoreceptors wearing the carboxyl tail of the alpha(1b) subtype had a high basal phosphorylation and displayed a strong phosphorylation in response to norepinephrine and phorbol esters. Our results demonstrate that stimulation of alpha(1a)-adrenergic receptor, or activation of protein kinase C, leads to alpha(1a)-adrenergic receptor phosphorylation. alpha(1a)-Adrenoreceptors are affected to a much lesser extent than alpha(1b)-adrenoreceptors by protein kinase C activation. PMID- 10692462 TI - Deletion of the COOH terminus converts the ST5 p70 protein from an inhibitor of RAS signaling to an activator with transforming activity in NIH-3T3 cells. AB - Expression of the human protein ST5-p70 correlates with reduced tumorigenic phenotype in mammalian cells, reverts their transformed phenotype, and restores their contact-dependent growth. Furthermore, expression of p70 in COS-7 cells suppresses activation of mitogen activated protein kinase MAPK/ERK2 by the largest ST5 product, p126, in response to epidermal growth factor stimulation. Here we show that deletions of the COOH-terminal region of p70 transform NIH3T3 cells and induce their anchorage-independent growth. Analysis of signaling leading to MAPK/ERK2 stimulation revealed that in COS-7 cells, expression of either p70-DeltaC1 or p70-DeltaC2 markedly enhanced ERK2 activity in a growth factor-independent manner. Whereas wild-type p70 slightly inhibited ERK2 activation by RAS and MEK2, co-expression or p70-DeltaC1 or p70-DeltaC2 with either protein stimulated ERK2 cooperatively. This activity was completely blocked by the dominant negative mutants RAS17N or MEKAA, suggesting that p70 functions upstream of RAS. Unlike wild-type p70, expression of p70-DeltaC1 or p70 DeltaC2 mutant did not interfere with the ability of ST5-p126 to stimulate ERK2. Taken together, the data suggest that the COOH-terminal tail, residues 489-609, contains some of the critical determinants for the function of p70. Loss of this region converts the protein from an inhibitor to a constitutive activator of the RAS-ERK2 pathway. PMID- 10692463 TI - Structural determinants required for apical sorting of an intestinal brush-border membrane protein. AB - The distinct protein and lipid constituents of the apical and basolateral membranes in polarized cells are sorted by specific signals. O-Glycosylation of a highly polarized intestinal brush-border protein sucrase isomaltase is implicated in its apical sorting through interaction with sphingolipid-cholesterol microdomains. We characterized the structural determinants required for this mechanism by focusing on two major domains in pro-SI, the membrane anchor and the Ser/Thr-rich stalk domain. Deletion mutants lacking either domain, pro SI(DeltaST) (stalk-free) and pro-SI(DeltaMA) (membrane anchor-free), were constructed and expressed in polarized Madin-Darby canine kidney cells. In the absence of the membrane anchoring domain, pro-SI(DeltaMA) does not associate with lipid rafts and the mutant is randomly delivered to both membranes. Therefore, the O-glycosylated stalk region is not sufficient per se for the high fidelity of apical sorting of pro-SI. Pro-SI(DeltaST) does not associate either with lipid rafts and its targeting behavior is similar to that of pro-SI(DeltaMA). Only wild type pro-SI containing both determinants, the stalk region and membrane anchor, associates with lipid microdomains and is targeted correctly to the apical membrane. However, not all sequences in the stalk region are required for apical sorting. Only O-glycosylation of a stretch of 12 amino acids (Ala(37)-Pro(48)) juxtapose the membrane anchor is required in conjunction with the membrane anchoring domain for correct targeting of pro-SI to the apical membrane. Other O glycosylated domains within the stalk (Ala(49)-Pro(57)) are not sufficient for apical sorting. We conclude that the recognition signal for apical sorting of pro SI comprises O-glycosylation of the Ala(37)-Pro(48) stretch and requires the presence of the membrane anchoring domain. PMID- 10692464 TI - Biosynthesis and post-translational processing of lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). N-linked glycosylation affects cell surface expression and ligand binding. AB - LOX-1 (lectin-like oxidized low density lipoprotein receptor-1) is a type II membrane protein belonging to the C-type lectin family that can act as a cell surface receptor for atherogenic oxidized low density lipoprotein (Ox-LDL) and may play crucial roles in atherogenesis. In this study, we show, by pulse-chase labeling and glycosidase digestion, that LOX-1 is synthesized as a 40-kDa precursor protein with N-linked high mannose carbohydrate chains (pre-LOX-1), which is subsequently further glycosylated and processed into the 48-kDa mature form within 40 min. Furthermore, when treated with an N-glycosylation inhibitor, tunicamycin, both tumor necrosis factor-alpha-activated bovine aortic endothelial cells and CHO-K1 cells stably expressing bovine LOX-1 (BLOX-1-CHO) exclusively produced a 32-kDa deglycosylated form of LOX-1. Cell enzyme-linked immunosorbent assay, flow cytometry, and immunofluorescence confocal microscopy demonstrated that the deglycosylated form of LOX-1 is not efficiently transported to the cell surface, but is retained in the endoplasmic reticulum or Golgi apparatus in tumor necrosis factor-alpha-activated bovine aortic endothelial cells, but not in BLOX 1-CHO cells. Radiolabeled Ox-LDL binding studies revealed that the deglycosylated form of LOX-1 expressed on the cell surface of BLOX-1-CHO cells has a reduced affinity for Ox-LDL binding. Taken together, N-linked glycosylation appears to play key roles in the cell-surface expression and ligand binding of LOX-1. PMID- 10692465 TI - Binding of factor VIIa to tissue factor on keratinocytes induces gene expression. AB - Binding of the zymogen serine protease Factor VII (FVII) to its cellular cofactor tissue factor (TF) triggers blood coagulation. Several recent reports have suggested that the formation of this complex may serve additional functions. We have used cDNA arrays to study differential gene expression in response to the interaction of activated FVII (FVIIa) with TF on a human keratinocyte cell line. Of 931 mRNA species observed up to 6 h after FVIIa (10 nM) addition, 24 were significantly up-regulated in what may resemble a wound-type response. Responders included mRNA species coding for transcription regulators (c-fos, egr-1, ETR101, BTEB2, c-myc, fra-1, and tristetraproline), growth factors (amphiregulin, hbEGF, CTGF, and FGF-5), proinflammatory cytokines (IL-1beta, IL-8, LIF, and MIP2alpha), proteins involved in cellular reorganization/migration (RhoE, uPAR, and collagenases 1 and 3), and others (PAI-2, cyclophilin, GADD45, Jagged1, and prostaglandin E(2) receptor). The transcriptional response to FVIIa was abrogated by antibodies to TF and left unaffected by hirudin. The pattern of genes induced suggests that the FVIIa.TF complex may play an active role in early wound repair as well as hemostasis. The former is a novel function ascribed to the complex that may also be contributing to the pathophysiology of unwarranted TF expression. PMID- 10692466 TI - Spatial requirements for 15-(R)-hydroxy-5Z,8Z,11Z, 13E-eicosatetraenoic acid synthesis within the cyclooxygenase active site of murine COX-2. Why acetylated COX-1 does not synthesize 15-(R)-hete. AB - The two isoforms of cyclooxygenase, COX-1 and COX-2, are acetylated by aspirin at Ser-530 and Ser-516, respectively, in the cyclooxygenase active site. Acetylated COX-2 is essentially a lipoxygenase, making 15-(R)-hydroxyeicosatetraenoic acid (15-HETE) and 11-(R)-hydroxyeicosatetraenoic acid (11-HETE), whereas acetylated COX-1 is unable to oxidize arachidonic acid to any products. Because the COX isoforms are structurally similar and share approximately 60% amino acid identity, we postulated that differences within the cyclooxygenase active sites must account for the inability of acetylated COX-1 to make 11- and 15-HETE. Residues Val-434, Arg-513, and Val-523 were predicted by comparison of the COX-1 and -2 crystal structures to account for spatial and flexibility differences observed between the COX isoforms. Site-directed mutagenesis of Val-434, Arg-513, and Val-523 in mouse COX-2 to their COX-1 equivalents resulted in abrogation of 11- and 15-HETE production after aspirin treatment, confirming the hypothesis that these residues are the major isoform selectivity determinants regulating HETE production. The ability of aspirin-treated R513H mCOX-2 to make 15-HETE, although in reduced amounts, indicates that this residue is not an alternate binding site for the carboxylate of arachidonate and that it is not the only specificity determinant regulating HETE production. Further experiments were undertaken to ascertain whether the steric bulk imparted by the acetyl moiety on Ser-530 prevented the omega-end of arachidonic acid from binding within the top channel cavity in mCOX-2. Site-directed mutagenesis was performed to change Val 228, which resides at the junction of the main cyclooxygenase channel and the top channel, and Gly-533, which is in the top channel. Both V228F and G533A produced wild type-like product profiles, but, upon acetylation, neither was able to make HETE products. This suggests that arachidonic acid orientates in a L-shaped binding configuration in the production of both prostaglandin and HETE products. PMID- 10692467 TI - Proteolytic processing and assembly of the C5 subunit into the proteasome complex. AB - Assembly of mammalian 20 S proteasomes from individual subunits is beginning to be investigated. Proteasomes are made of four heptameric rings in the configuration alpha7beta7beta7alpha7. By using anti-proteasome and anti-subunit specific antibodies, we characterized the processing and assembly of the beta subunit C5. The C5 precursor (25 kDa) remains as a free non-assembled polypeptide in the cell. The conversion of the C5 precursor to mature C5 (23 kDa) occurs concomitantly with its incorporation into 15 S proteasome intermediate and 20 S mature proteasome complexes. This processing is dependent on proteasome activity and takes place in the cytosol. These results are not fully compatible with the hypothesis that postulates that assembly of proteasomes takes place via a "half proteasome" intermediate that contains one full alpha-ring and one full beta-ring of unprocessed beta subunit precursors. PMID- 10692468 TI - Characterization of a repressor element and a juxtaposed tissue-restricted activator element located on the distal neu gene promoter. AB - The proto-oncogene neu (HER2 or c-erbB2) is overexpressed with or without gene amplification in 20-30% of breast cancers. In patients, neu amplification or overexpression in breast and ovarian cancer correlates with poor prognosis and tumor resistance to chemotherapy. neu-induced transformation can be reversed by the suppression of neu gene transcription. To further understand how neu gene transcription is regulated and to identify a possible transcriptional repressor(s) of neu, we identified a negative regulatory element known previously to be located within a 1-kilobase (kb) DNA fragment of an unknown sequence, upstream of the proximal neu gene promoter. One of several DNA fragments subcloned from this region suppressed transcriptional activity of the proximal neu gene promoter. Sequencing of the 1-kb fragment confirmed the location of the repressor element to be between an AluI and a RsaI sites, around 1.4 kb upstream to the translation start site. Various deletions were introduced into the AluI RsaI fragment and subcloned into both the native neu promoter and a heterologous thymidine kinase promoter. Subsequent transfections and reporter gene assays in cell lines of various tissues of origin confirmed and narrowed the repressor activity to a 120-base pair NlaIV-MslI fragment located between -1385 and -1266. Importantly, specific protein binding activity to this element could be detected with nuclear extracts isolated from these cell lines. In contrast, a 28-base pair MslI-RsaI fragment (-1265 to -1238), located immediately 3' of the putative repressor element, was found to form protein-DNA complexes with only nuclear extracts isolated from a colon carcinoma cell line. This specific protein binding activity correlated with a previously unknown transcriptional stimulatory activity only in this cell line. PMID- 10692469 TI - Synergy of SF1 and RAR in activation of Oct-3/4 promoter. AB - The Oct-3/4 transcription factor is expressed in the earliest stages of embryogenesis, and is thus likely to play an important role in regulation of initial decisions in development. For the first time, we have shown that SF1 and Oct-3/4 are co-expressed in embryonal carcinoma (EC) P19 cells, and their expression is down-regulated with very similar kinetics following retinoic acid (RA) induced differentiation of these cells, suggesting a functional relationship between the two. Previously, we have shown that the Oct-3/4 promoter harbors an RA-responsive element, RAREoct, which functions in EC cells as a binding site for positive regulators of transcription, such as RAR and RXR. In this study we have identified in the Oct-3/4 promoter two novel SF1-binding sites: SF1(a) and SF1(b). The proximal site, SF1(a), is located within the RAREoct, and the distal site, SF1(b), is located between nucleotide -193 and -209 of the Oct-3/4 promoter. Both sites contribute to activation of Oct-3/4 promoter in EC cells, with SF1(a) playing a more crucial role. SF1, and its isoforms ELP2 and ELP3 bind to both SF1 sites and activate the Oct-3/4 promoter. This activation depends on the presence of SF1 DNA-binding domain. Thus, Oct-3/4 is the first EC-specific gene reported that is regulated by SF1. Interestingly, SF1 and RAR form a novel complex on the RAREoct sequence that synergistically activate the Oct-3/4 promoter. Both RARE and SF1 cis regulatory elements, as well as the SF1 DNA binding domain, are needed for this synergism. SF1 and Oct-3/4 transcription factors play a role in the same developmental regulatory cascade. PMID- 10692470 TI - Inherited defects of sodium-dependent glutamate transport mediated by glutamate/aspartate transporter in canine red cells due to a decreased level of transporter protein expression. AB - Canine red cells have a high affinity Na(+)/K(+)-dependent glutamate transporter. We herein demonstrate that this transport is mediated by the canine homologue of glutamate/aspartate transporter (GLAST), one of the glutamate transporter subtypes abundant in the central nervous system. We also demonstrate that GLAST is the most ubiquitous glutamate transporter among the transporter subtypes that have been cloned to date. The GLAST protein content was extremely reduced in variant red cells, low glutamate transport (LGlut) red cells characterized by an inherited remarkable decrease in glutamate transport activity. All LGluT dogs carried a missense mutation of Gly(492) to Ser (G492S) in either the heterozygous or homozygous state. The GLAST protein with G492S mutation was fully functional in glutamate transport in Xenopus oocytes. However, G492S GLAST exhibited a marked decrease in activity after the addition of cycloheximide, while the wild type showed no significant change, indicating that G492S GLAST was unstable compared with the wild-type transporter. Moreover, LGluT dogs, but not normal dogs, heterozygous for the G492S mutation showed a selective decrease in the accumulation of GLAST mRNA from the normal allele. Based on these findings, we conclude that a complicated heterologous combination of G492S mutation and some transcriptional defect contributes to the pathogenesis of the LGluT red cell phenotype. PMID- 10692471 TI - Transforming growth factor-beta1 enhances Ha-ras-induced expression of cyclooxygenase-2 in intestinal epithelial cells via stabilization of mRNA. AB - Oncogenic ras induces the expression of cyclooxygenase-2 (COX-2) in a variety of cells. Here we investigated the role of transforming growth factor-beta (TGF beta) in the Ras-mediated induction of COX-2 in intestinal epithelial cells (RIE 1). RIE-1 cells were transfected with an inducible Ha-Ras(Val12) cDNA and are referred as RIE-iRas cells. the addition of 5 mM isopropyl-1-thio-beta-D galactopyranoside (IPTG) induced the expression of Ha-Ras(Val12), closely followed by an increase in the expression of COX-2. Neutralizing anti-TGF-beta antibody partially blocked the Ras-induced increase in COX-2. Combined treatment with IPTG and TGF-beta1 resulted in a 20-50-fold increase in the levels of COX-2 mRNA. The t1/2 of COX-2 mRNA was increased from 13 to 24 min by Ha-Ras induction alone. The addition of TGF-beta1 further stabilized the COX-2 mRNA (t1/2 > 50 min). Stable transfection of a luciferase reporter construct containing the COX-2 3'-untranslated region (3'-UTR) revealed that TGF-beta1 treatment and Ras induction each stabilized the COX-2 3'-UTR. Combined treatment with IPTG and TGF beta1 synergistically increased the luciferase activity. Furthermore, a conserved AU-rich region located in the proximal COX-2 3'-UTR is required for maximal stabilization of COX-2 3'-UTR by Ras or TGF-beta1 and is necessary for the synergistic stabilization of COX-2 3'-UTR by oncogenic Ras and TGF-beta1. PMID- 10692472 TI - The insect salivary protein, prolixin-S, inhibits factor IXa generation and Xase complex formation in the blood coagulation pathway. AB - Prolixin-S is a salivary anticoagulant of the blood-sucking insect, Rhodnius prolixus, and known as an inhibitor of the intrinsic Xase. We report here its inhibitory mechanisms with additional important anticoagulation activities. We found prolixin-S specifically bound to factor IX/IXa in the presence of Ca(2+) ions. Light scattering and surface plasmon resonance studies showed that prolixin S interfered with factor IX/IXa binding to the phospholipid membrane, indicating that prolixin-S inhibit Xase activity of factor IXa by interference with its Xase complex formation. Furthermore, reconstitution experiments showed that prolixin-S binding to factor IX strongly inhibited factor IXa generation by factor XIa. We also found that prolixin-S inhibited factor IXa generation by factor VIIa-tissue factor complex and factor IXalpha generation by factor Xa. These results suggest that prolixin-S inhibits both intrinsic and extrinsic coagulations by sequential inhibition of all coagulation pathways in which factor IX participates. It was also suggested that prolixin-S may bind to factor IX/IXa by recognizing conformational change of the Gla domain induced by Ca(2+) binding. PMID- 10692473 TI - Correlated oscillations in glucose consumption, oxygen consumption, and intracellular free Ca(2+) in single islets of Langerhans. AB - Micron-sized sensors were used to monitor glucose and oxygen levels in the extracellular space of single islets of Langerhans in real-time. At 10 mM glucose, oscillations in intraislet glucose concentration were readily detected. Changes in glucose level correspond to changes in glucose consumption by glycolysis balanced by mass transport into the islet. Oscillations had a period of 3.1 +/- 0.2 min and amplitude of 0.8 +/- 0.1 mM glucose (n = 21). Superimposed on these oscillations were faster fluctuations in glucose level during the periods of low glucose consumption. Oxygen level oscillations that were out of phase with the glucose oscillations were also detected. Oscillations in both oxygen and glucose consumption were strongly dependent upon extracellular Ca(2+) and sensitive to nifedipine. Simultaneous measurements of glucose with intracellular Ca(2+) ([Ca(2+)](i)) revealed that decreases in [Ca(2+)](i) preceded increases in glucose consumption by 7.4 +/- 2.1 s during an oscillation (n = 9). Conversely, increases in [Ca(2+)](i) preceded increases in oxygen consumption by 1.5 +/- 0.2 s (n = 4). These results suggest that during oscillations, bursts of glycolysis begin after Ca(2+) has stopped entering the cell. Glycolysis stimulates further Ca(2+) entry, which in turn stimulates increases in respiration. The data during oscillation are in contrast to the time course of events during initial exposure to glucose. Under these conditions, a burst of oxygen consumption precedes the initial rise in [Ca(2+)](i). A model to explain these results is described. PMID- 10692474 TI - Ionizing radiation exposure results in up-regulation of Ku70 via a p53/ataxia telangiectasia-mutated protein-dependent mechanism. AB - Genome damaging events, such as gamma-irradiation exposure, result in the induction of pathways that activate DNA repair mechanisms, halt cell cycle progression, and/or trigger apoptosis. We have investigated the effects of gamma irradiation on cellular levels of the Ku autoantigens. Ku70 and Ku80 have been shown to form a heterodimeric complex that can bind tightly to free DNA ends and activate the protein kinase DNA-PKcs. We have found that irradiation results in an up-regulation of cellular levels of Ku70, but not Ku80, and that this enhanced level of Ku70 accumulates within the nucleus. Further, we uncovered that the postirradiation up-regulation of Ku70 utilizes a mechanism that is dependent on both p53 and damage response protein kinase ATM (ataxia-telangiectasia-mutated); however, the activation of DNA-PK does not require Ku70 up-regulation. These findings suggest that Ku70 up-regulation provides the cell with a means of assuring either proper DNA repair or an appropriate response to DNA damage independent of DNA-PKcs activation. PMID- 10692475 TI - Upstream tissue inhibitor of metalloproteinases-1 (TIMP-1) element-1, a novel and essential regulatory DNA motif in the human TIMP-1 gene promoter, directly interacts with a 30-kDa nuclear protein. AB - Elevated expression of the tissue inhibitor of metalloproteinases-1 (TIMP-1) protein and mRNA has been reported in human diseases including cancers and tissue fibrosis. Regulation of TIMP-1 gene expression is mainly mediated at the level of gene transcription and involves the activation of several well known transcription factors including those belonging to the AP-1, STAT, and Pea3/Ets families. In the current study, we have used DNase-1 footprinting to identify a new regulatory element (5'-TGTGGTTTCCG-3') present in the human TIMP-1 gene promoter. Mutagenesis and transfection studies in culture-activated rat hepatic stellate cells and the human Jurkat T cell line demonstrated that the new element named upstream TIMP-1 element-1 (UTE-1) is essential for transcriptional activity of the human TIMP-1 promoter. Electrophoretic mobility shift assay studies revealed that UTE-1 can form protein-DNA complexes of distinct mobilities with nuclear extracts from a variety of mammalian cell types and showed that induction of a high mobility UTE-1 complex is associated with culture activation of freshly isolated rat hepatic stellate cells. A combination of UV-cross-linking and Southwestern blotting techniques demonstrated that UTE-1 directly interacts with a 30-kDa nuclear protein that appears to be present in all cell types tested. We conclude that UTE-1 is a novel regulatory element that in combination with its cellular binding proteins may be an important component of the mechanisms controlling TIMP-1 expression in normal and pathological states. PMID- 10692477 TI - Functional expression of a multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster and its C-terminal deletion mutants. AB - The occurrence of a deoxyribonucleoside kinase in Drosophila melanogaster (Dm dNK) with remarkably broad substrate specificity has recently been indicated (Munch-Petersen, B., Piskur, J., and Sondergaard, L. (1998) J. Biol. Chem. 273, 3926-3931). To prove that the capacity to phosphorylate all four deoxyribonucleosides is in fact associated to one polypeptide chain, partially sequenced cDNA clones, originating from the Berkeley Drosophila genome sequencing project, were searched for homology with human deoxyribonucleoside kinases. The total sequence of one cDNA clone and the corresponding genomic DNA was determined and expressed in Escherichia coli as a glutathione S-transferase fusion protein. The purified and thrombin cleaved recombinant protein phosphorylated the four deoxyribonucleosides with high turnover and K(m) values similar to those of the native Dm-dNK, as well as the four ribonucleosides and many therapeutical nucleoside analogs. Dm-dNK has apparently the same origin as the mammalian kinases, thymidine kinase 2, deoxycytidine kinase, deoxyguanosine kinase, and the herpes viral thymidine kinases, but it has a unique C terminus that seems to be important for catalytic activity and specificity. The C-terminal 20 amino acids were dispensable for phosphorylation of deoxyribonucleosides but necessary for full activity with purine ribonucleosides. Removal of the C-terminal 20 amino acids increased the specific activity 2-fold, but 99% of the activity was lost after removal of the C-terminal 30 amino acids. PMID- 10692476 TI - Characterization of alpha-crystallin-plasma membrane binding. AB - Alpha-crystallin, a large lenticular protein complex made up of two related subunits (alphaA- and alphaB-crystallin), is known to associate increasingly with fiber cell plasma membranes with age and/or the onset of cataract. To understand better the binding mechanism, we developed a sensitive membrane binding assay using lens plasma membranes and recombinant human alphaA- and alphaB-crystallins conjugated to a small fluorescent tag (Alexa350). Both alphaA and alphaB homopolymer complexes, as well as a reconstituted 3:1 heteromeric complex, bind to lens membranes in a specific, saturable, and partially irreversible manner that is sensitive to both time and temperature. The amount of alpha-crystallin that binds to the membrane increases under acidic pH conditions and upon removal of exposed intrinsic membrane protein domains but is not affected at high ionic strength, suggesting that alpha-crystallin binds to the fiber cell plasma membranes mainly through hydrophobic interactions. The binding capacity and affinity for the reconstituted 3:1 heteromeric complex were measured to be 3. 45 +/- 0.11 ng/microg of membrane and 4.57 +/- 0.50 x 10(-4) microg(-1) of membrane, respectively. The present membrane binding data support the hypothesis that the physical properties of a mixed alpha-crystallin complex may hold particular relevance for the function of alpha-crystallin within the lens. PMID- 10692478 TI - Expression and function of calcium binding domain chimeras of the integrins alpha(IIb) and alpha(5). AB - To further identify amino acid domains involved in the ligand binding specificity of alpha(IIb)beta(3), chimeras of the conserved calcium binding domains of alpha(IIb) and the alpha subunit of the fibronectin receptor alpha(5)beta(1) were constructed. Chimeras that replaced all four calcium binding domains, replaced all but the second calcium binding domain of alpha(IIb) with those of alpha(5), or deleted all four calcium binding domains were synthesized but not expressed on the cell surface. Additional chimeras exchanged subsets or all of the variant amino acids in the second calcium binding domain, a region implicated in ligand binding. Cell surface expression of each second calcium binding domain mutant complexed with beta(3) was observed. Each second calcium binding domain mutant was able to 1) bind to immobilized fibrinogen, 2) form fibrinogen-dependent aggregates after treatment with dithiothreitol, and 3) bind the activation dependent antibody PAC1 after LIBS 6 treatment. Soluble fibrinogen binding studies suggested that there were only small changes in either the K(d) or B(max) of any mutant. We conclude that chimeras of alpha(IIb) containing the second calcium binding domain sequences of alpha(5) are capable of complexing with beta(3), that the complexes are expressed on the cell surface, and that mutant complexes are capable of binding both immobilized and soluble fibrinogen, suggesting that the second calcium binding domain does not determine ligand binding specificity. PMID- 10692479 TI - Cloning, expression, and substrate specificity of a fungal chymotrypsin. Evidence for lateral gene transfer from an actinomycete bacterium. AB - Unlike trypsins, chymotrypsins have not until now been found in fungi. Expressed sequence tag analysis of the deuteromycete Metarhizium anisopliae identified two trypsins (family S1) and a novel chymotrypsin (CHY1). CHY1 resembles actinomycete (bacterial) chymotrypsins (family S2) rather than other eukaryote enzymes (family S1) in being synthesized as a precursor species (374 amino acids, pI/MW: 5.07/38,279) containing a large N-terminal fragment (186 amino acids). Chy1 was expressed in Pichia pastoris yielding an enzyme with a chymotrypsin specificity for branched aliphatic and aromatic C-terminal amino acids. This is predictable as key catalytic residues determining the specificity of Streptomyces griseus chymotrypsins are conserved with CHY1. Mature (secreted) CHY1 (pI/MW: 8.29/18,499) shows closest overall amino acid identity to S. griseus protease C (55%) and clustered with other secreted bacterial S2 chymotrypsins that diverged widely from animal and endocellular bacterial enzymes in phylogenetic trees of the chymotrypsin superfamily. Conversely, actinomycete chymotrypsins are much more closely related to fungal proteases than to other eubacterial sequences. Complete genomes of yeast, gram eubacteria, archaebacteria, and mitochondria do not contain paralogous genes. Expressed sequence tag data bases from other fungi also lack chymotrypsin homologs. In light of this patchy distribution, we conclude that chy1 probably arose by lateral gene transfer from an actinomycete bacterium. PMID- 10692480 TI - Ca(2+) requirement for high-affinity gamma-aminobutyric acid (GABA) binding at GABA(B) receptors: involvement of serine 269 of the GABA(B)R1 subunit. AB - The gamma-aminobutyric acid (GABA) receptor type B (GABA(B)R) is constituted of at least two homologous proteins, GABA(B)R1 and GABA(B)R2. These proteins share sequence and structural similarity with metabotropic glutamate and Ca(2+)-sensing receptors, both of which are sensitive to Ca(2+). Using rat brain membranes, we report here that the affinity of GABA and 3-aminopropylphosphinic acid for the GABA(B)R receptor is decreased by a factor >10 in the absence of Ca(2+). Such a large effect of Ca(2+) is not observed with baclofen or the antagonists CGP64213 and CGP56999A. In contrast to baclofen, the potency of GABA in stimulating GTPgammaS binding in rat brain membranes is also decreased by a factor >10 upon Ca(2+) removal. The potency for Ca(2+) in regulating GABA affinity was 37 microM. In cells expressing GABA(B)R1, the potency of GABA, but not of baclofen, in displacing bound (125)I-CGP64213 was similarly decreased in the absence of Ca(2+). To identify residues that are responsible for the Ca(2+) effect, the pharmacological profile and the Ca(2+) sensitivity of a series of GABA(B)R1 mutants were examined. The mutation of Ser269 into Ala was found to decrease the affinity of GABA, but not of baclofen, and the GABA affinity was found not to be affected upon Ca(2+) removal. Finally, the effect of Ca(2+) on the GABA(B) receptor function is no longer observed in cells coexpressing this GABA(B)R1 S269A mutant and the wild-type GABA(B)R2. Taken together, these results show that Ser269, which is conserved in the GABA(B)R1 protein from Caenorhabditis elegans to mammals, is critical for the Ca(2+)-effect on the heteromeric GABA(B) receptor. PMID- 10692481 TI - Structure-activity relationship of staurosporine analogs in regulating expression of endothelial nitric-oxide synthase gene. AB - In human umbilical vein endothelial cells and in human umbilical vein endothelial cell-derived EA.hy 926 cells, staurosporine (Stsp) and its glycosidic indolocarbazole analogs 7-hydroxystaurosporine (UCN-01) and 4'-N-benzoyl staurosporine (CGP 41251) enhanced nitric-oxide synthase (NOS) III mRNA expression (analyzed by RNase protection assay), protein expression (determined by Western blot), and activity [measured by rat fetal lung fibroblast (RFL-6) reporter cell assay] in a concentration- and time-dependent manner. In contrast, the bisindolylmaleimide analogs GF 109203X, Ro 31-8220 and Go 6983 had no effect on NOS III expression, and Go 6976, a methyl- and cyanoalkyl-substituted nonglycosidic indolocarbazole derivative of Stsp, even reduced NOS III expression in a concentration-dependent fashion. The up-regulation of NOS III expression by Stsp and analogs appears to be a transcriptional event because Stsp, 7 hydroxystaurosporine, and CGP 41251 enhanced the activity of a 1.6-kb human NOS III promoter fragment transiently transfected into EA.hy 926 endothelial cells. Stsp and analogs did not affect the stability of the NOS III mRNA. Stsp is known as a potent protein kinase (PK) inhibitor. Data obtained with other kinase inhibitors (and stimulators) indicated, however, that the effect of Stsp and analogs on NOS III expression was unrelated to the activities of PKC, PKA, PKG, or tyrosine kinase(s). Stsp analogs such as CGP 41251 also counteracted the NOS III mRNA-decreasing effect of tumor necrosis factor-alpha. These findings demonstrate that Stsp analogs represent a new class of compounds positively interacting with the transcription of the endothelial NOS III gene. Such compounds may prove useful in the prophylaxis and therapy of vascular disease. PMID- 10692482 TI - Allosteric interactions between the antagonist prazosin and amiloride analogs at the human alpha(1A)-adrenergic receptor. AB - It has been demonstrated previously that amilorides can interact with a well defined allosteric site on the human alpha(2A)-adrenergic receptor. In this study, the question was explored as to whether the human alpha(1A)-adrenergic receptor also possesses an equivalent allosteric site. The six amilorides examined strongly increased the dissociation rate of the antagonist [(3)H]prazosin from the alpha(1A)-adrenergic receptor in a concentration dependent manner. With the parent amiloride, the dissociation data were well fitted by an equation derived from the ternary complex allosteric model, compatible with amiloride acting at a defined allosteric site on the alpha(1A) adrenergic receptor. In contrast, the dissociation data for [(3)H]prazosin in the presence of the amiloride analogs were not compatible with the equation derived from a one-allosteric-site model, but could be fitted well by an equation derived from a two-allosteric-site model. However, certain individual parameters could not be resolved. The observed dissociation rate constants increased steeply with increasing amiloride analog concentration, and in some cases the data could be fitted with a logistic equation. The slope factors calculated from such fits were 1.2 to 2.1. It is concluded that the structure-binding relationships of the amilorides at the alpha(1A)- and alpha(2A)-adrenergic receptors are different. The interactions of the five amiloride analogs, but not the parent amiloride, with the alpha(1A)-adrenergic receptor are compatible with the presence of two (but not one) allosteric sites, and is thus more complex than that found for the alpha(2A)-adrenergic receptor. PMID- 10692483 TI - Modulation of dopamine D(2) receptor signaling by actin-binding protein (ABP 280). AB - Proteins that bind to G protein-coupled receptors have recently been identified as regulators of receptor anchoring and signaling. In this study, actin-binding protein 280 (ABP-280), a widely expressed cytoskeleton-associated protein that plays an important role in regulating cell morphology and motility, was found to associate with the third cytoplasmic loop of dopamine D(2) receptors. The specificity of this interaction was originally identified in a yeast two-hybrid screen and confirmed by protein binding. The functional significance of the D(2) receptor-ABP-280 association was evaluated in human melanoma cells lacking ABP 280. D(2) receptor agonists were less potent in inhibiting forskolin-stimulated cAMP production in these cells. Maximal inhibitory responses of D(2) receptor activation were also reduced. Further yeast two-hybrid experiments showed that ABP-280 association is critically dependent on the carboxyl domain of the D(2) receptor third cytoplasmic loop, where there is a potential serine phosphorylation site (S358). Serine 358 was replaced with aspartic acid to mimic the effects of receptor phosphorylation. This mutant (D(2)S358D) displayed compromised binding to ABP-280 and coupling to adenylate cyclase. PKC activation also generated D(2) receptor signaling attenuation, but only in ABP-containing cells, suggesting a PKC regulatory role in D(2)-ABP association. A mechanism for these results may be derived from a role of ABP-280 in the clustering of D(2) receptors, as determined by immunocytochemical analysis in ABP-deficient and replete cells. Our results suggest a new molecular mechanism of modulating D(2) receptor signaling by cytoskeletal protein interaction. PMID- 10692484 TI - Characterization of protein kinase chk1 essential for the cell cycle checkpoint after exposure of human head and neck carcinoma A253 cells to a novel topoisomerase I inhibitor BNP1350. AB - Cellular topoisomerase I is an important target in cancer chemotherapy. A novel karenitecin, BNP1350, is a topoisomerase I-targeting anticancer agent with significant antitumor activity against human head and neck carcinoma A253 cells in vitro. As a basis for future clinical trials of BNP1350 in human head and neck carcinoma, in vitro studies were carried out to investigate its effect on DNA damage and cell cycle checkpoint response. The treatment of A253 cells with BNP1350 caused biphasic profiles of DNA fragmentation displayed from 0 to 48 h after 2-h exposure. Pulsed-field gel electrophoresis demonstrated that the first wave of DNA damage was mainly megabase DNA fragmentation, but the second wave of DNA damage was 50- to 300-kb DNA fragmentation in addition to megabase DNA damage. The cell cycle checkpoint response was characterized after exposure to 0.07 and 0.7 microM concentrations of BNP1350, the IC(50) and IC(90) values, respectively. After exposure to a low concentration of BNP1350 (IC(50)), A253 cells accumulated primarily in G(2) phase. In contrast, treatment with a high concentration of BNP1350 (IC(90)) resulted in S phase accumulation. The concentration-associated cell cycle perturbation by BNP1350 was correlated with different profiles of cell cycle-regulatory protein expression. When treated with the low concentration of BNP1350, cyclin B/cdc2 protein expression was up regulated, whereas with the high concentration, no significant change was observed at 24 and 48 h. In addition, increased phosphorylation of a G(2) checkpoint kinase chk1 was observed when cells were treated with a low concentration of BNP1350, whereas only slight inhibition of chk1 activity was found in the cells treated with the higher concentration. Altered chk1 phosphorylation after DNA damage appears to be associated with specific phases of cell cycle arrest induced by BNP1350. Because A253 cells do not express the p53 protein, the drug-induced alterations of the G(2) checkpoint kinase chk1 are not p53-dependent. PMID- 10692485 TI - Specific regulation of RGS2 messenger RNA by angiotensin II in cultured vascular smooth muscle cells. AB - The effects of angiotensin II (Ang II) are mediated primarily by Ang II type 1 receptors, which in turn are coupled to heterotrimeric G proteins. After receptor activation, the G(alpha) and G(betagamma) subunits dissociate, contributing to the signaling cascades involving protein kinase C (PKC) activation. Regulators of G protein signaling (RGS proteins) comprise a class of proteins that have been shown to negatively regulate the G(alpha) subunit. We examined which RGS sequences were expressed in vascular smooth muscle cells and which of these were regulated by Ang II. Reverse transcription-polymerase chain reaction showed that of 16 RGS sequences screened, six RGS transcripts (RGS2, 3, 10, 11, and 12 and GAIP) were present. Northern blot analysis demonstrated that RGS3, 10, and 12 and GAIP were not regulated by Ang II at the mRNA level. In contrast, RGS2 mRNA was rapidly and dose dependently increased (395 +/- 24% peak, 45 min) by Ang II but returned to baseline level by 6 to 8 h. Phorbol-12-myristate-13-acetate, a PKC activator, robustly increased RGS2. This signal was attenuated by the PKC inhibitor GF 109203X (50 +/- 4%) and by phorbol-12, 13-dibutyrate-mediated down regulation of PKC (48 +/- 13%). Tyrosine kinase inhibition and calcium deprivation did not affect the up-regulation of RGS2 mRNA after Ang II stimulation. Actinomycin D treatment inhibited both Ang II- and phorbol-12 myristate-13-acetate-stimulated RGS2 up-regulation, suggesting activation of transcription by these agonists. The stability of RGS2 mRNA did not appear to be affected by Ang II. Thus, RGS2 is a likely candidate for negative regulation of the G proteins coupled to the Ang II type 1 receptor in vascular smooth muscle cells. Regulation of this protein may be of critical importance in modulating the role of Ang II in vascular disease. PMID- 10692486 TI - Substrate-dependent regulation of human arylamine N-acetyltransferase-1 in cultured cells. AB - Arylamine N-acetyltransferase-1 (NAT1) is a polymorphically expressed enzyme that is widely distributed throughout the body. In the present study, we provide evidence for substrate-dependent regulation of this enzyme. Human peripheral blood mononuclear cells cultured in medium supplemented with p-aminobenzoic acid (PABA; 6 microM) for 24 h showed a significant decrease (50-80%) in NAT1 activity. The loss of activity was concentration-dependent (EC(50) approximately 2 microM) and selective because PABA had no effect on the activity of constitutively expressed lactate dehydrogenase or aspartate aminotransferase. PABA also induced down-regulation of NAT1 activity in several human cell lines grown at confluence. Substrate-dependent down-regulation was not restricted to PABA. Addition of other NAT1 substrates, such as p-aminosalicylic acid, ethyl-p aminobenzoate, or p-aminophenol to peripheral blood mononuclear cells in culture also resulted in significant (P <.05) decreases in NAT1 activity. However, addition of the NAT2-selective substrates sulfamethazine, dapsone, or procainamide did not alter NAT1 activity. Western blot analysis using a NAT1 specific antibody showed that the loss of NAT1 activity was associated with a parallel reduction in the amount of NAT1 protein (r(2) = 0.95). Arylamines that did not decrease NAT1 activity did not alter NAT1 protein levels. Semiquantitative reverse transcriptase polymerase chain reaction of mRNA isolated from treated and untreated cells revealed no effect of PABA on NAT1 mRNA levels. We conclude that NAT1 can be down-regulated by arylamines that are themselves NAT1 substrates. Because NAT1 is involved in the detoxification/activation of various drugs and carcinogens, substrate-dependent regulation may have important consequences with regard to drug toxicity and cancer risk. PMID- 10692487 TI - A single glycine residue at the entrance to the first membrane-spanning domain of the gamma-aminobutyric acid type A receptor beta(2) subunit affects allosteric sensitivity to GABA and anesthetics. AB - Site-directed mutagenesis of the gamma-aminobutyric acid type A (GABA(A)) receptor beta(2) subunit has demonstrated that conversion of a conserved glycine residue located at the entrance to the first transmembrane domain into the homologous rho(1) residue phenylalanine alters the modulating effects of four different i.v. anesthetics: pentobarbital, alphaxalone, etomidate, and propofol. Using the baculovirus expression system in Spodoptera frugiperda 9 cells, anesthetic-induced enhancement of [(3)H]muscimol and [(3)H]flunitrazepam binding in receptors containing the beta(2)(G219F) point mutation displayed a significantly reduced efficacy in modulation by all four i.v. anesthetics tested. Furthermore, GABA(A) receptors containing the alpha(1)(G223F) point mutation also significantly decreased the maximal effect of etomidate- and propofol-induced enhancement of ligand binding. Conversely, the homologous point mutation in rho(1) receptors (F261G) changed the i.v. anesthetic-insensitive receptor to confer anesthetic modulation of [(3)H]muscimol binding. Consistent with the binding, functional analysis of pentobarbital-enhanced GABA currents recorded with whole-cell patch clamp demonstrated the beta(2)(G219F) subunit mutation eliminated the potentiating effect of the anesthetic. Similarly, propofol enhanced GABA currents were potentiated less in alpha(1)beta(2)(G219F)gamma(2) receptors than in alpha(1)beta(2)gamma(2) receptors. Although ligand binding displayed comparable K(D) values for muscimol among wild-type, alpha(1)beta(2)gamma(2), and mutant receptors, patch-clamp recordings showed that alpha(1)beta(2)(G219F)gamma(2) receptors had a significantly more potent response to GABA than did alpha(1)beta(2)gamma(2) or alpha(1)(G223F)beta(2)gamma(2). The alpha(1)beta(2)(G219F)gamma(2) receptors also were more sensitive to direct channel activation by pentobarbital and propofol in the absence of GABA. These results suggest that the first transmembrane glycine residue on the beta(2) subunit may be important for conformational or allosteric interactions of channel gating by both GABA and anesthetics. PMID- 10692488 TI - Inhibition of c-myc expression in cells by targeting an RNA-protein interaction using antisense oligonucleotides. AB - Antisense oligodeoxynucleotides (ODNs) are designed to bind to and inhibit a target mRNA. We used a novel approach for the design of ODNs to the c-myc mRNA using protein binding sites as targets for ODN action. Our strategy was to identify ODNs that could interfere with the coding region determinant-binding protein (CRD-BP), a protein that binds to the CRD region of the c-myc mRNA. Using an in vitro gel shift assay, we show that ODN molecules can occlude the CRD-BP from the mRNA. The best ODN, CRD-ODN4, was able to inhibit RNA binding of the CRD BP by 75%. This effect was sequence-specific and concentration dependent. K562 cells treated with a 2'-O-methyl derivative of CRD-ODN4 showed a concentration dependent decrease in both c-myc mRNA and protein levels, with a maximal 65% inhibition of protein expression at 200 nM CRD-ODN4. In contrast, a 2'-O-methyl ODN derivative targeting the translation initiation codon (antimyc-aug) reduced c myc protein but actually increased mRNA levels, an effect resulting at least partly from stabilization of the c-myc mRNA. CRD-ODN4 treatment did not alter the c-myc mRNA half-life. CRD-ODN4 was more effective in inhibiting K562 cell growth than antimyc-aug, reducing cell number by approximately 70% after 48 h of exposure to 750 nM. The correlation between ODN effects on RNA-protein interactions in vitro and those observed in cells supports the hypothesis that CRD-ODN4 inhibits the interaction between the CRD-BP and the c-myc mRNA and that disrupting this RNA-protein interaction reduces c-myc expression in cells. PMID- 10692489 TI - Functional inactivation of the nociceptin receptor by alanine substitution of glutamine 286 at the C terminus of transmembrane segment VI: evidence from a site directed mutagenesis study of the ORL1 receptor transmembrane-binding domain. AB - A site-directed mutagenesis approach has been used to gain insight into the molecular events whereby the heptadecapeptide nociceptin binds and activates the opioid receptor-like 1 (ORL1) receptor, a G protein-coupled receptor. Alanine mutation, in the human ORL1 receptor, of transmembrane amino acid residues that are conserved in opioid receptors, Asp(130) and Tyr(131) in transmembrane segment (TM) III, Phe(220) and Phe(224) in TM V, and Trp(276) in TM VI, yields mutant receptors with reduced affinity, and proportionally decreased reactivity, toward nociceptin. Least to most deleterious in this respect are Ala substitutions of Phe(220) approximately W276A < Tyr(131) << Phe(224) 10,000 nM compared with 0.8 nM at the wild-type receptor). In all respects, this mutant receptor appears to be functionally inactive, indicating that residue Gln(286) may play a pivotal role in ORL1 receptor-mediated transduction of the nociceptin signal. PMID- 10692490 TI - Effect of p53 status on sensitivity to platinum complexes in a human ovarian cancer cell line. AB - Wild-type p53 is frequently mutated in late-stage ovarian cancer and has been proposed as a determinant of cisplatin chemosensitivity. We have therefore established a human ovarian cancer cell line differing only in p53 status and characterized its response after treatment with different platinum complexes. The wild-type p53-expressing cell line A2780 was stably transfected with HPV-16 E6 (E6) or an empty vector (VC) as control. Parental A2780 and VC had similar cisplatin sensitivities, whereas E6 was 3- to 4-fold more sensitive as measured by sulforhodamine B and clonogenic assay. E6 was 2- to 3-fold more sensitive to transplatin and the novel cisplatin analog ZD0473 than VC, whereas the trans platinum analog JM335 was approximately equitoxic. Platinum uptake was similar for all of the cell lines after cisplatin. The removal of platinum-DNA adducts, as measured by atomic absorption spectroscopy, was reduced in E6 compared with VC after cisplatin but similar after JM335. After 10 microM cisplatin, the G(1) population (0-96 h) was reduced in E6 cells compared with VC, whereas the S phase (8-48 h) and G(2) phase (48-96 h) were increased. Similar proportions of VC and E6 cells died by apoptosis, as detected by annexin V binding, but more E6 cells died by necrosis relative to VC. Our results suggest that the loss of functional p53 can increase cisplatin cytotoxicity in A2780, with loss of G(1)/S checkpoint control and decreased cisplatin-DNA adduct repair, but these effects can be circumvented by the use of JM335, which forms different DNA-platinum adducts. PMID- 10692491 TI - Hematotoxicity of the chinese herbal medicine Tripterygium wilfordii hook f in CD34-positive human bone marrow cells. AB - T2, a chloroform/methanol extract of the herb Tripterygium wilfordii Hook f, has been used in China for the treatment of autoimmune and inflammatory diseases for many years. Recent experimental evidence has confirmed that T2 has potent anti inflammatory and immunosuppressive activity, and a United States Food and Drug Administration-approved clinical trial is currently exploring the efficacy of T2 in the treatment of rheumatoid arthritis. Despite the potential therapeutic benefits of T2, there is ample documentation that T2 is toxic, targeting, among other things, the hematopoietic system, and its use has resulted in cases of leukopenia, thrombocytopenia, and aplastic anemia. This investigation was undertaken to characterize the in vitro effects of T2 on primary human CD34 positive (CD34+) bone marrow cells. Our results demonstrate that T2 has a potent inhibitory effect on the clonogenic response of human bone marrow cells to exogenously added hematopoietic growth factors. The inhibition of colony formation by T2 is not the result of direct cytotoxicity or increased apoptosis and indicates a functional suppression of hematopoiesis. Additional experiments demonstrate that T2 also alters transcriptional regulation in bone marrow cells by inhibiting nuclear factor-kappaB. This transcription factor is found in CD34+ bone marrow cells and has been recently shown to be a requirement for colony formation. These results demonstrate that therapeutic concentrations of T2 exert a significant hematotoxic effect by inhibiting growth factor response in CD34+ bone marrow cells and suggest that inhibition of nuclear factor-kappaB may play a role in the blood dyscrasias encountered with the use of this drug. PMID- 10692492 TI - Calcium-independent receptor for alpha-latrotoxin and neurexin 1alpha [corrected] facilitate toxin-induced channel formation: evidence that channel formation results from tethering of toxin to membrane. AB - alpha-Latrotoxin binding to the calcium-independent receptor for alpha-latrotoxin (CIRL-1), a putative G-protein-coupled receptor, stimulates secretion from chromaffin and PC12 cells. Using patch clamp techniques and microspectrofluorimetry, we demonstrate that the interaction of alpha-latrotoxin with CIRL-1 produces a high conductance channel that permits increases in cytosolic Ca(2+). alpha-Latrotoxin interaction with CIRL-1 transiently expressed in bovine chromaffin cells produced a 400-pS channel, which rarely closed under Ca(2+)-free conditions. The major effect of overexpressing CIRL-1 was to greatly increase the sensitivity of chromaffin cells to channel formation by alpha latrotoxin. alpha-Latrotoxin interaction with CIRL-1 transiently overexpressed in non-neuronal human embryonic kidney 293 (HEK293) cells produced channels that were nearly identical with those observed in chromaffin cells. Channel currents were reduced by millimolar Ca(2+). At alpha-latrotoxin concentrations below 500 pM, channel formation occurred many seconds after binding of toxin to CIRL-1 indicating distinct steps in channel formation. In all cases there was a rapid, sequential addition of channels once the first channel appeared. An analysis of CIRL-1 mutants indicated that channel formation in HEK293 cells is unlikely to be transduced by a G-protein-dependent mechanism. alpha-Latrotoxin interaction with a fusion construct composed of the extracellular domain of CIRL-1 anchored to the membrane by the transmembrane domain of vesicular stomatitis virus glycoprotein, and with neurexin 1alpha, an alpha-latrotoxin receptor structurally unrelated to CIRL-1, produced channels virtually identical with those observed with wild-type CIRL-1. We propose that alpha-latrotoxin receptors recruit toxin to facilitate its insertion across the membrane and that alpha-latrotoxin itself controls the conductance properties of the channels it produces. PMID- 10692493 TI - Effect of 6-aminonicotinamide and other protein synthesis inhibitors on formation of platinum-DNA adducts and cisplatin sensitivity. AB - The present study was undertaken to examine the mechanistic basis for the recent observation that the pyridine nucleotide derivative 6-aminonicotinamide (6AN, NSC 21206) enhances the accumulation and resulting cytotoxicity of cisplatin in a variety of tumor cell lines. When A549 lung cancer cells or K562 leukemia cells were treated with 62.5 microM 6AN for 21 h and then pulse-labeled with [(35)S]methionine for 1 h, increased labeling of five polypeptides, one of which corresponded to a M(r) approximately 78,000 glucose-regulated protein (GRP78), was observed. Two subsequent observations, however, suggested that up-regulation of these polypeptides was unlikely to explain the interaction between 6AN and cisplatin: 1) the concentration of 6AN required to induce GRP78 was 4-fold higher than the dose required to sensitize cells to cisplatin; and 2) simultaneous treatment of cells with 6AN and cycloheximide prevented the increase in GRP78 but not the sensitizing effect of 6AN. On the contrary, treatment with the protein synthesis inhibitors cycloheximide, anisomycin, or puromycin as well as prolonged exposure to the RNA synthesis inhibitor actinomycin D mimicked the biochemical modulating effects of 6AN on cisplatin action. Conversely, 6AN inhibited protein synthesis, whereas 18 6AN analogs that failed to enhance Pt-DNA adducts and cisplatin cytotoxicity failed to inhibit protein synthesis. These observations are consistent with a model in which 6AN and other inhibitors of protein synthesis act as modulating agents by increasing cisplatin accumulation, thereby enhancing the formation of Pt-DNA adducts and subsequent cisplatin-induced cell death. PMID- 10692494 TI - Identification of threonine residues controlling the agonist-dependent phosphorylation and desensitization of the rat A(3) adenosine receptor. AB - Activation of the A(3) adenosine receptor (A(3)AR) contributes to the cardioprotective, bronchoconstrictive, and hypotensive effects of adenosine. Agonist occupation of the A(3)AR results in a rapid desensitization of receptor function, which is associated with the phosphorylation of the receptor protein by one or more members of the G protein-coupled receptor kinase family of protein kinases. Although we demonstrated previously that phosphorylation of the C terminal 14 amino acids of the rat A(3)AR is crucial for rapid desensitization to occur, the identity of the critical phosphorylation sites has remained unknown. Here, we demonstrate that the simultaneous mutation of Thr(307), Thr(318), and Thr(319) to Ala residues dramatically reduces agonist-stimulated phosphorylation and rapid desensitization of the rat A(3)AR. Individual mutation of each residue demonstrated that Thr(318) and Thr(319) are the major sites of phosphorylation. Phosphorylation at Thr(318) appeared to be necessary to observe phosphorylation at Thr(319), but not vice versa. However, the replacement of Thr(318) with a glutamate residue demonstrated that the simple addition of negative charge at position 318 was not sufficient to rescue phosphorylation at position 319. In addition, the mutation of two predicted palmitoylation-site cysteine residues proximal to the regulatory domain resulted in the appearance of an agonist independent basal phosphorylation. Therefore, G protein-coupled receptor kinase mediated phosphorylation of the C-terminal tail of the A(3)AR in situ appears to follow a sequential mechanism, perhaps involving receptor depalmitoylation, with phosphorylation at Thr(318) being particularly important. PMID- 10692495 TI - Subtype-specific kinetics of inhibitory adenosine receptor internalization are determined by sensitivity to phosphorylation by G protein-coupled receptor kinases. AB - Despite coupling to the same class of inhibitory G proteins and binding the same physiological ligand, the human A(1) and rat A(3) adenosine receptors (ARs) desensitize at different rates in response to sustained agonist exposure. This is due to the ability of the A(3)AR, but not the A(1)AR, to serve as a substrate for rapid phosphorylation and desensitization by members of the G protein-coupled receptor kinase (GRK) family. The aim of this study was to investigate whether these differences were also manifested in their abilities to undergo agonist dependent receptor internalization. For the first time, we report that A(3)ARs internalize profoundly in response to short-term exposure to agonist but not activators of second messenger-regulated kinases. The A(3)AR-selective antagonist MRS1523 blocked both A(3)AR phosphorylation and internalization. Moreover, in contrast to the A(1)AR, which internalized quite slowly (t(1/2) = 90 min), A(3)ARs internalized rapidly (t(1/2) = 10 min) over a time frame that followed the onset of receptor phosphorylation. A nonphosphorylated A(3)AR mutant failed to internalize over a 60-min time course, suggesting that receptor phosphorylation was essential for rapid A(3)AR internalization to occur. In addition, fusion onto the A(1)AR of the A(3)AR C-terminal domain containing the sites for phosphorylation by GRKs conferred rapid agonist-induced internalization kinetics (t(1/2) = 10 min) on the resulting chimeric AR. In conclusion, these data suggest that GRK-stimulated phosphorylation of threonine residues within the C-terminal domain of the A(3)AR is obligatory to observe rapid agonist-mediated internalization of the receptor. PMID- 10692497 TI - Calmodulin increases the sensitivity of type 3 inositol-1,4, 5-trisphosphate receptors to Ca(2+) inhibition in human bronchial mucosal cells. AB - Inositol-1,4,5-trisphosphate (IP(3)) releases Ca(2+) from intracellular stores by binding to its receptor (IP(3)R), a multigene family of Ca(2+)-release channels consisting of IP(3)R1, IP(3)R2, and IP(3)R3. IP(3)R1 is stimulated by low cytoplasmic Ca(2+) concentrations and inhibited by high concentrations. Discrepant reports appeared about the effect of cytoplasmic Ca(2+) on IP(3)R3, showing either a bell-shaped dependence or only a stimulatory phase with no negative feedback by high Ca(2+) concentrations. We investigated how calmodulin interfered with the feedback of cytosolic Ca(2+) on the unidirectional IP(3) induced Ca(2+) release in permeabilized 16HBE14o- bronchial mucosal cells, where IP(3)R3 represents 93% of the receptors at the mRNA level and 81% at the protein level. Calmodulin inhibited the Ca(2+) release induced by 1.5 microM IP(3) with an IC(50) value of 9 microM. This inhibition was absolutely dependent on the presence of cytosolic Ca(2+). Ca(2+) inhibited the IP(3)R with an IC(50) value of 0.92 microM Ca(2+) in the absence of calmodulin and with an IC(50) value of 0.15 microM Ca(2+) in its presence. It is concluded that: 1) IP(3)R3 can be inhibited by calmodulin, 2) IP(3)R3 is inhibited by high Ca(2+) concentrations, and 3) calmodulin shifts the inhibitory part of the Ca(2+)-response curve toward lower Ca(2+) concentrations. PMID- 10692496 TI - RPR112378 and RPR115781: two representatives of a new family of microtubule assembly inhibitors. AB - A screening program aimed at the discovery of new antimicrotubule agents yielded RPR112378 and RPR115781, two natural compounds extracted from the Indian plant Ottelia alismoides. We report their isolation, structural determination, and mechanisms of action. RPR112378 is an efficient inhibitor of tubulin polymerization (IC(50) = 1.2 microM) and is able to disassemble preformed microtubules. Regarding tubulin activity, RPR115781 is 5-fold less active than RPR112378. Tubulin-RPR112378 complexes, when isolated by gel filtration, were able to block further tubulin addition to growing microtubules, a mechanism that accounts for the substoichiometric effect of the drug. RPR112378 was found to prevent colchicine binding but not vinblastine binding to tubulin. Although colchicine binding is known to induce an increase of tubulin GTPase activity, no such increase was observed with RPR112378. We show that RPR112378 is a highly cytotoxic compound and that RPR115781 is 10, 000-fold less active as an inhibitor of KB cell growth. Part of the cytotoxicity of RPR112378 is probably caused by a reaction of addition with sulfhydryl groups, an observation that has not been made with RPR115781. In conclusion, these molecules represent a new class of inhibitors of microtubule assembly with potential therapeutic value. PMID- 10692498 TI - A steroid derivative with paclitaxel-like effects on tubulin polymerization. AB - The endogenous estrogen metabolite 2-methoxyestradiol has modest antimitotic activity that may result from a weak interaction at the colchicine binding site of tubulin, but it nevertheless has in vivo antitumor activity. Synthetic efforts to improve activity led to compounds that increased inhibitory effects on cell growth, tubulin polymerization, and binding of colchicine to tubulin. This earlier work was directed at modifications in the steroid A ring, which is probably analogous to the colchicine tropolonic C ring. One of the most active analogs prepared was 2-ethoxyestradiol (2EE). We report here that different modifications in the steroid B ring of 2EE yield compounds with two apparently distinct modes of action. Simple expansion of the B ring to seven members resulted in a compound comparable to 2EE in its ability to inhibit tubulin polymerization and colchicine binding to tubulin. Acetylation of the hydroxyl groups in this analog and in 2EE essentially abolished these inhibitory properties. The introduction of a ketone functionality at C6, together with acetylation of the hydroxyls at positions 3 and 17, produced a compound with activity similar to that of paclitaxel, in that the agent enhanced tubulin polymerization into polymers that were partially stable at 0 degrees C. The acetyl group at C17, but not that at C3, was essential for this paclitaxel-like activity. PMID- 10692499 TI - Cloned human aquaporin-1 is a cyclic GMP-gated ion channel. AB - Aquaporin-1 (AQP1) is a member of the membrane intrinsic protein (MIP) gene family and is known to provide pathways for water flux across cell membranes. We show here that cloned human AQP1 not only mediates water flux but also serves as a cGMP-gated ion channel. Two-electrode voltage-clamp analyses showed consistent activation of an ionic conductance in wild-type AQP1-expressing oocytes after the direct injection of cGMP (50 nl of 100 mM). Current activation was not observed in control (water-injected) oocytes or in AQP5-expressing oocytes with osmotic water permeabilities equivalent to those seen with AQP1. Patch-clamp recordings revealed large conductance channels (150 pS in K(+) saline) in excised patches from AQP1-expressing oocytes after the application of cGMP to the internal side. Amino acid sequence alignments between AQP1 and sensory cyclic-nucleotide-gated channels showed similarities between the cyclic-nucleotide-gated binding domain and the AQP1 carboxyl terminus that were not present in AQP5. Competitive radioligand-binding assays with [(3)H]cGMP demonstrated specific binding (K(D) = 0.2 microM) in AQP1-expressing Sf9 cells but not in controls. These results indicate that AQP1 channels have the capacity to participate in ionic signaling after the activation of cGMP second-messenger pathways. PMID- 10692500 TI - Catechol-O-methyltransferase inhibition attenuates levodopa toxicity in mesencephalic dopamine neurons. AB - Inhibition of catechol-O-methyltransferase (COMT; EC 2.1.1.6) is a new therapeutic strategy in the treatment of Parkinson's disease. However, nothing is known about the effects of COMT inhibition on levodopa (L-dopa)-induced toxicity in dopamine (DA) neurons. Therefore we evaluated the effects of the selective COMT inhibitors Ro 41-0960, OR-486, and tolcapone alone and in combination with L dopa in primary mesencephalic cultures from rat. Neither COMT inhibitor affected the growth of tyrosine hydroxylase immunoreactive (THir) cells with concentrations up to 10 microM when studied alone. However, Ro 41-0960 reduced the L-dopa-induced THir cell loss after 24 h in a dose-dependent manner, shifting the TD(50) value from 21 microM in the absence to 71 microM in the presence of 1 microM Ro 41-0960 (P <.01) without affecting survival of non-DA neurons. OR-486 and the clinically used COMT inhibitor tolcapone showed similar effects. In contrast, toxicity induced by D-dopa was not altered by COMT inhibitors. Furthermore, the primary metabolite of L-dopa formed by COMT, 3-O-methyldopa, and the methyl group donor S-adenosyl-L-methionine used by COMT did not alter THir neuron survival and L-dopa-induced toxicity, respectively, with concentrations up to 100 microM. These data demonstrate that COMT inhibition attenuates L-dopa toxicity toward DA neurons in vitro, but probably not by preventing 3-O methyldopa production or cellular S-adenosyl-L-methionine depletion. PMID- 10692501 TI - myo-inositol 1,4,6-trisphosphorothioate and myo-inositol 1,3, 6 trisphosphorothioate: partial agonists with very low intrinsic activity at the platelet myo-inositol 1,4,5-trisphosphate receptor. AB - Racemic mixtures and enantiomerically pure D-isomers of both myo-inositol 1,3,6 trisphosphorothioate [Ins(1,3,6)PS(3)] and myo-inositol 1,4,6 trisphosphorothioate [Ins(1,4,6)PS(3)], prepared by total synthesis, were examined in Ca(2+) flux and binding assays. Both D-Ins(1,3,6)PS(3) and D Ins(1,4,6)PS(3) were shown to be low intrinsic activity partial agonists at the platelet myo-inositol 1,4, 5-trisphosphate [Ins(1,4,5)P(3)] receptor, releasing less than 20% of the Ins(1,4,5)P(3)-sensitive Ca(2+) store. D-Ins(1,4,6)PS(3) displaced specifically bound [(3)H]Ins(1,4,5)P(3) from rat cerebellar membranes, although displacement was some 34-fold weaker than by D-Ins(1,4,5)P(3). D Ins(1,4,6)PS(3) displaced [(3)H]Ins(1,4, 5)P(3) from cerebellar membranes with roughly twice the affinity of DL-Ins(1,4,6)PS(3) (IC(50) value = 1.4 +/- 0.35 microM compared with 2.15 +/- 0.13 microM), whereas D-Ins(1,3,6)PS(3) displaced [(3)H]Ins(1,4,5)P(3) with roughly twice the affinity of DL-Ins(1,3, 6)PS(3) (IC(50) value = 17.5 +/- 5.8 microM compared with 34 +/- 10 microM), confirming that the activity of both these phosphorothioates resides in their D-enantiomers. Increasing concentrations of either D-Ins(1,3,6)PS(3) or D-Ins(1,4,6)PS(3) were able to partially antagonize Ca(2+) release induced by submaximal concentrations of Ins(1,4,5)P(3), an inhibition that could be overcome by increasing the concentration of Ins(1,4,5)P(3), suggesting competition for binding at the Ins(1,4,5)P(3)-R. The only low-efficacy partial agonists at the Ins(1,4,5)P(3)-R discovered to date have been phosphorothioates; the novel D-Ins(1,3,6)PS(3) and D Ins(1,4,6)PS(3) can now be added to this small group of analogs. However, D Ins(1,4,6)PS(3) has a relatively high affinity for the Ins(1,4,5)P(3)-R but maintains the lowest efficacy of all the partial agonists thus far identified. As such, it may be a useful tool for pharmacological intervention in the polyphosphoinositide pathway and an important lead compound for the development of further Ins(1,4,5)P(3)-R antagonists. PMID- 10692502 TI - Potency of ligands correlates with affinity measured against agonist and inverse agonists but not against neutral ligand in constitutively active chemokine receptor. AB - ORF-74, a 7TM receptor oncogene encoded by human herpes virus 8, shows 50% constitutive activity in stimulating phosphatidylinositol turnover and binds a large variety of CXC chemokines. These endogenous ligands cover a full spectrum of pharmacological properties with growth-related oncogene (GRO)-alpha and -gamma functioning as full agonists; GRObeta as a partial agonist; interleukin (IL)-8, neutrophil-activating peptide (NAP)-2, and epithelial cell-derived activating peptide (ENA)-78 as neutral ligands; granulocyte colony-stimulating factor (GCP) 2 as a partial inverse agonist; and interferon-gamma inducible protein (IP)-10 and stromal cell-derived factor (SDF)-1alpha as full inverse agonists. The affinity for the agonists was independent of whether it was determined in competition binding against the agonist (125)I-GROalpha, against the inverse agonist (125)I-IP-10, or against the neutral ligand (125)I-IL-8. Similarly, the affinities of the inverse agonists were within 1 order of magnitude independent of the choice of radioligand. In contrast, the neutral ligands IL-8, NAP-2, and ENA-78, which all displaced (125)I-IL-8 with single-digit nanomolar affinity showed up to 1000-fold lower affinity against both the radioactive agonist and against the radioactive inverse agonist. A close correlation was observed between the EC(50) values for the ligands and their IC(50) values measured against either radioactive agonist or radioactive inverse agonist, but a poor correlation was found to the IC(50) value measured against the neutral ligand. It is concluded that in ORF-74, ligands compete for binding more according to pharmacological property than to structural homology and that both agonists and inverse agonists, in contrast to neutral ligands, apparently bind with high affinity either to a common conformation of the receptor or to readily interconvertible states, not available for the neutral ligands. PMID- 10692503 TI - Transcriptional induction of heme oxygenase-1 gene expression by okadaic acid in primary rat hepatocyte cultures. AB - Heme oxygenase (HO) catalyzes the rate-limiting enzymatic step of heme degradation and regulates the cellular heme content. The gene expression of the inducible isoform of HO, HO-1, is up-regulated in response to various agents causing oxidative stress. To investigate the regulatory role of protein phosphatases in the hepatic regulation of HO-1 gene expression, primary cultures of rat hepatocytes were treated with okadaic acid (OA), which specifically inhibits the serine threonine protein phosphatases 1 and 2A. Both protein synthesis and mRNA expression of HO-1 were induced by OA in cultured hepatocytes, but not in cultured tissue macrophages of rat liver. The HO-1 mRNA induction by OA occurred in a time- and concentration-dependent manner. Simultaneous treatment with OA plus dibutyryl cAMP caused a synergistic up-regulation of steady-state levels of HO-1 mRNA, and the specific protein kinase A inhibitor KT5720 markedly reduced the OA-dependent HO-1 mRNA induction. In contrast, the dibutyryl cAMP dependent induction of the phosphoenolpyruvate carboxykinase mRNA expression and enzyme activity was inhibited by simultaneous treatment with OA in hepatocytes. The induction of the HO-1 gene expression by OA was transcriptional as determined by studies with actinomycin D, nuclear run-off assay, and measurement of the half life of HO-1 mRNA. Luciferase reporter constructs containing DNA sequences of the rat HO-1 promoter 5'-flanking region were up-regulated by OA in transiently transfected hepatocytes. Mutation of the cAMP response element/activator protein 1 (-665/-654) site obliterated the OA-dependent induction, suggesting that this element is involved in the transcriptional induction of the rat HO-1 gene by OA. PMID- 10692504 TI - The human glutathione transferase P1-1 specific inhibitor TER 117 designed for overcoming cytostatic-drug resistance is also a strong inhibitor of glyoxalase I. AB - gamma-L-Glutamyl-S-(benzyl)-L-cysteinyl-R-(-)-phenylglycine (TER 117) has previously been developed for selective inhibition of human glutathione S transferase P1-1 (GST P1-1) based on the postulated contribution of this isoenzyme to the development of drug resistance in cancer cells. In the present investigation, the inhibitory effect of TER 117 on the human glyoxalase system was studied. Although designed as an inhibitor specific for GST P1-1, TER 117 also competitively inhibits glyoxalase I (K(I) = 0.56 microM). In contrast, no inhibition of glyoxalase II was detected. Reduced glyoxalase activity is expected to raise intracellular levels of toxic 2-oxoaldehydes otherwise eliminated by glyoxalase I. The resulting toxicity would accompany the potentiation of cytostatic drugs, caused by inhibition of the detoxication effected by GST P1-1. TER 117 was designed for efficient inhibition of the most abundant form GST P1 1/Ile105. Therefore, the inhibitory effect of TER 117 on a second allelic variant GST P1-1/Val105 was also studied. TER 117 was shown to competitively inhibit both GST P1-1 variants. The apparent K(I) values at glutathione concentrations relevant to the intracellular milieu were in the micromolar range for both enzyme forms. Extrapolation to free enzyme produced K(I) values of approximately 0.1 microM for both isoenzymes, reflecting the high affinity of GST P1-1 for the inhibitor. Thus, the allelic variation in position 105 of GST P1-1 does not affect the inhibitory potency of TER 117. The inhibitory effects of TER 117 on GST P1-1 and glyoxalase I activities may act in synergy in the cell and improve the effectiveness of chemotherapy. PMID- 10692505 TI - Functional importance of transmembrane helix 6 Trp(279) and exoloop 3 Val(299) of rat gonadotropin-releasing hormone receptor. AB - Previous studies have established that the interaction of gonadotropin-releasing hormone (GnRH) with its receptor (GnRHR) would require partial entry of the N- and C-terminal regions of ligand into the transmembrane core. The functional significance of the conserved aromatic residue Trp(279) present in the transmembrane helix 6, and Val(299) located in exoloop 3 of the rat GnRHR was investigated by mutagenesis followed by expression in Chinese hamster ovary-K1 cells. Compared with wild-type, substitution of Trp(279) with Ser or Arg resulted in a marked reduction or total abolition, respectively, of ligand binding and, in both cases, abrogation of GnRH-induced inositol phosphate production. A total absence of functionality was observed when Val(299) was simply replaced with Ala. Mention should be made that an expression of all mutated and wild-type receptor proteins was observed. Interestingly, the double mutant [Trp(279)Arg/Val(299)Ala]GnRHR restored B(max) to wild type (504 +/- 43 versus 541 +/- 41 fmol/mg protein), but with a diminished affinity (4.95 +/- 1.05 versus 0.94 +/- 0.35 nM), and GnRH failed to induce inositol phosphate. No influence of the mutations was seen on internalization of the receptor. The three-dimensional model of GnRH binding to the rat GnRHR was built predicting that Trp(279) is buried at 20 A in the transmembrane core of the receptor, directly in contact with Trp(3) of GnRH. In contrast, Val(299) is located in a region that cannot be precisely defined at the extracellular end of transmembrane helix 7. Although models cannot provide any clue concerning the observed interactivity between the two distal residues, altogether these data reveal the functional importance of both GnRHR Trp(279) and Val(299) and suggest that Trp(279), interacting with GnRH Trp(3), represents the bottom of the binding pocket. PMID- 10692506 TI - Transport function and hepatocellular localization of mrp6 in rat liver. AB - The multidrug resistance-associated proteins (Mrps) constitute a family of cellular export pumps of the ATP-binding cassette transporter superfamily and play an important role in hepatobiliary excretion. We investigated the transport function and subcellular localization of mrp6, a novel member of the mrp family, in rat liver. Transport studies in vesicles isolated from mrp6 expressing Sf9 cells identified the anionic cyclopentapeptide and endothelin receptor antagonist BQ-123 as a substrate of mrp6 (K(m) approximately 17 microM). Besides BQ-123, which is also a substrate of mrp2 (K(m) approximately 124 microM), no other common substrates were found for mrp2, mrp6, and the canalicular bile salt export pump Bsep. The cyclic peptides endothelin I and Arg(8)-vasopressin were transported by mrp2 but not by mrp6. Using a polyclonal antiserum raised against a C-terminal peptide, mrp6 was found to be localized at the lateral and, to a lesser extent, at the canalicular plasma membrane of hepatocytes. The limited overlap of the substrate specificity with the canalicular export pumps mrp2 and Bsep indicates that mrp6 does not play a major role in canalicular organic anion excretion. However, its dual localization at the lateral and canalicular plasma membrane suggests that mrp6 might fulfill a "housekeeping" transport function involved in the regulation of paracellular and/or transcellular solute movement from blood into bile. PMID- 10692507 TI - 5-Iodo-A-85380, an alpha4beta2 subtype-selective ligand for nicotinic acetylcholine receptors. AB - In an effort to develop selective radioligands for in vivo imaging of neuronal nicotinic acetylcholine receptors (nAChRs), we synthesized 5-iodo-3-(2(S) azetidinylmethoxy)pyridine (5-iodo-A-85380) and labeled it with (125)I and (123)I. Here we present the results of experiments characterizing this radioiodinated ligand in vitro. The affinity of 5-[(125)I]iodo-A-85380 for alpha4beta2 nAChRs in rat and human brain is defined by K(d) values of 10 and 12 pM, respectively, similar to that of epibatidine (8 pM). In contrast to epibatidine, however, 5-iodo-A-85380 is more selective in binding to the alpha4beta2 subtype than to other nAChR subtypes. In rat adrenal glands, 5-iodo-A 85380 binds to nAChRs containing alpha3 and beta4 subunits with 1/1000th the affinity of epibatidine, and exhibits 1/60th and 1/190th the affinity of epibatidine at alpha7 and muscle-type nAChRs, respectively. Moreover, unlike epibatidine and cytisine, 5-[(125)I]iodo-A-85380 shows no binding in any brain regions in mice homozygous for a mutation in the beta2 subunit of nAChRs. Binding of 5-[(125)I]iodo-A-85380 in rat brain is reversible, and is characterized by high specificity and a slow rate of dissociation of the receptor-ligand complex (t(1/2) for dissociation approximately 2 h). These properties, along with other features observed previously in in vivo experiments (low toxicity, rapid penetration of the blood-brain barrier, and a high ratio of specific to nonspecific binding), suggest that this compound, labeled with (125)I or (123)I, is superior to other radioligands available for in vitro and in vivo studies of alpha4beta2 nAChRs, respectively. PMID- 10692508 TI - Adrenomedullin as a renal regulator peptide. PMID- 10692509 TI - Connective tissue growth factor: just another factor in renal fibrosis? PMID- 10692510 TI - Cocaine use and kidney damage. PMID- 10692511 TI - Multiple myeloma and renal failure. PMID- 10692512 TI - At which stage of renal failure should dialysis be started? PMID- 10692513 TI - Comparison of survival on CAPD and haemodialysis: statistical pitfalls. PMID- 10692514 TI - Renal failure following cardiac transplantation. PMID- 10692515 TI - Should renal transplantation be offered to older patients? PMID- 10692516 TI - Tubular cell damage in acute renal failure-apoptosis, necrosis, or both. PMID- 10692517 TI - BK-virus nephropathy in renal transplants-tubular necrosis, MHC-class II expression and rejection in a puzzling game. AB - We review BK-virus nephropathy (BKN) as a new complication that increasingly affects renal allografts and causes dysfunction. Since starting in 1996, we have seen 11 cases. Currently, the prevalence of BKN is 3% in our graft biopsies. The diagnosis can only be made histologically. The virus affects tubular epithelial cells that show characteristic intranuclear inclusion bodies. The major reason for impaired graft function and a possible way for viral particles to gain access to the blood via peritubular capillaries is necrosis of infected epithelial cells. BK-virus DNA in the plasma, which can be detected by PCR, is closely associated with nephropathy. BK-virus does not stimulate tubular MHC-class II expression as judged by immunofluorescence double labelling. The inflammatory response is inconsistent and the frequency of rejection episodes is not increased during disease. Clinical manifestation of viral nephropathy evolves in several stages. (i) Initial, asymptomatic and reversible activation of the virus, judged by the presence of inclusion bearing cells in the urine. (ii) High dose immunosuppressive drug regimens, often including tacrolimus. (iii) Tubular injury and viraemia as additional promoting conditions. BKN nephropathy was associated with graft loss in 45% of our patients. The remaining patients with persistent viral nephropathy showed renal dysfunction (serum creatinine levels on average 150% above baseline readings). Currently, no established antiviral therapy is available. We discuss attempts to lower immunosuppression as a means to control viral replication. We propose a diagnostic algorithm for screening and monitoring the disease. PMID- 10692518 TI - In the queue for a cadaver donor kidney transplant: new rules and concepts in the Eurotransplant International Foundation. PMID- 10692519 TI - CyA and OxLDL cause endothelial dysfunction in isolated arteries through endothelin-mediated stimulation of O(2)(-) formation. AB - BACKGROUND: Cyclosporin A (CyA) and oxidized low-density lipoprotein (OxLDL) cause endothelial dysfunction, partly through stimulation of O(2)(-) formation (which can inactivate nitric oxide). We investigated whether CyA and OxLDL potentiate their influence on oxidative stress, whether endothelin (ET) is a mediator of CyA- and OxLDL-induced O(2)(-) formation, and whether enhanced oxidative stress results in further attenuation of endothelium-dependent vasodilation. METHODS AND RESULTS: Human LDL was oxidized by Cu(++). O(2)(-) formation of isolated rat aortic rings was measured using a chemiluminescence assay. Incubation (60 min) of aortic rings with CyA (10 ng-10 microg/ml) or with OxLDL (300 microg/ml) caused a significant, dosedependent increase of the basal O(2)(-) formation. Pretreatment of the aortic rings with CyA (10 ng/ml) further enhanced the OxLDL-induced O(2)(-) formation by factor 1.9. The enhancement of the OxLDL-induced stimulation of O(2)(-) formation by CyA could be completely blocked by BQ123, a selective endothelin-1 (ET-1) receptor antagonist. Likewise, exogenously applied ET-1 (1 nM) potentiated the OxLDL-induced O(2)(-) formation by factor 1.8. Endothelium-dependent dilation was measured in isolated rings of rabbit aorta superfused with physiological salt solution in an organ bath. Incubation of the aortic rings with CyA (10 microg/ml, 60 min) or with OxLDL (300 microg/ml, 60 min) alone did not attenuate endothelium-dependent dilations. However, coincubation of the aortic rings with CyA+OxLDL in the presence of diethyl-dithio-carbamate, an inhibitor of the endogenous superoxide dismutase, caused a 60% inhibition of acetylcholine-induced dilator responses. CONCLUSIONS: Coincubation of isolated aortic rings with CyA and OxLDL causes a potent enhancement of vascular O(2)(-) formation. ET-1 seems to be mediator of the CyA induced O(2)(-) formation. Enhanced oxidative stress results in further attenuation of endothelium dependent vasodilation. PMID- 10692520 TI - Aminoguanidine ameliorates changes in the IGF system in experimental diabetic nephropathy. AB - BACKGROUND: Formation of advanced glycation end-products (AGEs) has been implicated in the development of diabetic complications. As well as causing changes in structural proteins, AGEs may also alter gene expression of growth factors in vitro. The insulin-like growth factor (IGF) system, including IGF-I and modulatory IGF binding proteins (IGFBPs), is dysregulated during the development of diabetic nephropathy. METHODS: Quantitative in situ hybridization histochemistry and immunohistochemistry were used to determine the effects of aminoguanidine, an inhibitor of AGE formation, on gene expression of IGF-I and IGFBPs in kidneys of long-term (8 months duration) streptozotocin-diabetic rats. RESULTS: Diabetes was associated with increased renal expression of IGFBP-1 mRNA (diabetes 824+/-236 vs control 264+/-76 arbitrary units, P<0.01) and decreased expression of mRNAs for IGF-I (diabetes 39+/-7 vs control 185+/-23 arbitrary units, P<0.001) and IGFBP-4 (diabetes 139+/-25 vs control 383+/-54 arbitrary units, P<0.001). Aminoguanidine treatment inhibited the effects of diabetes on renal expression of mRNA for IGF-I, IGFBP-1 and IGFBP-4. The changes in IGF-I and IGFBP-1 mRNA levels were reflected in altered peptide levels. In diabetic kidneys, IGFBP-5 mRNA levels were slightly decreased to 75% of control levels (P<0.01); aminoguanidine had no effect on IGFBP-5 mRNA levels. CONCLUSIONS: These results suggest that amelioration of changes in the renal IGF system by aminoguanidine may contribute to the renoprotective effects of the latter, which have been previously shown to inhibit structural and functional aspects of diabetic nephropathy in the rat. PMID- 10692521 TI - Glycoxidative modification of AA amyloid deposits in renal tissue. AB - BACKGROUND: N(epsilon)-carboxymethyllysine (CML) is a product of the oxidative modification of glycated proteins, which damages proteins with ageing, diabetes, uraemia and Alzheimer's disease. In contrast, pyrraline is one of the advanced glycation end products, which is independent of oxidative processes. CML has been identified in beta-amyloid of Alzheimer's disease and beta(2)-microglobulin associated amyloid. We investigated whether CML and pyrraline are formed in AA and AL amyloid of the kidney. METHOD: Renal specimens from 19 cases of AA amyloidosis and 14 cases of AL amyloidosis were investigated for immunolocalization of CML, pyrraline, collagen type IV and laminin in amyloid deposits. Renal biopsies of 10 age-matched cases with thin basement membrane disease and normal renal function were used as controls. The fractional areas of amyloid, CML, laminin and collagen IV in glomeruli and interstitium (%amyloid, %CML, %laminin and %collagen, respectively) were calculated using the point counting method. The correlation between these parameters was evaluated using Spearman's rank correlation test. RESULTS: CML colocalized with AA amyloid, but not AL amyloid, except in two cases of the latter with a long history of nephropathy exceeding 14 years. In contrast, pyrraline was not observed in either type of amyloid. Mean %CML in AA amyloid was significantly higher than %collagen and %laminin in glomeruli and interstitium, indicating that AA amyloid is modified by CML independent of colocalized extracellular matrix. %CML significantly correlated with %amyloid both in glomeruli and interstitium in AA amyloidosis. AL amyloid cases with a long history of nephropathy showed positive staining for CML in glomeruli and interstitium but no staining for collagen IV and laminin in amyloid deposits. CONCLUSION: CML modification may occur in amyloid deposits of AA amyloidosis, independent of extracellular matrix components. Glycoxidative modification may have a functional link to AA amyloid deposition in renal tissues. PMID- 10692522 TI - Citrate determines calcium oxalate crystallization kinetics and crystal morphology-studies in the presence of Tamm-Horsfall protein of a healthy subject and a severely recurrent calcium stone former. AB - BACKGROUND: The aim of this study was to measure the effects of normal (nTHP) and abnormal stone former Tamm-Horsfall protein (SF-THP) on calcium oxalate (CaOx) nucleation and aggregation as well as on crystal morphology, in presence or absence of citrate. METHODS: Nucleation and aggregation of CaOx crystals from a supersaturated, stirred solution (200 mM NaCl, 10 mM Na-acetate, pH 5.70, 5 mM Ca and 0.5 mM Ox) were studied by spectrophotometric time-course measurements of OD at 620 nm (OD(620)). Measured parameters were induction time t(I) (time to induce formation of detectable particles), S(N), (slope of increase of OD(620), mainly due to crystal nucleation), and S(A), (slope of decrease of OD(620) after equilibrium has been reached, due to crystal aggregation). Effects of citrate, nTHP and SF-THP on these parameters were measured, and scanning electron microscopy (SEM) was performed. RESULTS: At 1.5, 2.5 and 3.5 mM, citrate increased t(I) and inhibited crystal nucleation (by 78-87%) as well as aggregation (by 63-70%), and smaller CaOx crystals (length/width ratio 1.7+/-0.1) than under standard conditions (length/width 3.9+/-0.5) were visible (P<0.001). Normal THP at 30 and 40 mg/l inhibited crystal nucleation and, more strongly, aggregation (inhibition 76-81%). SEM revealed a decrease in length/width ratio to 2.6+/-0.4 (P=0.051 vs standard conditions) and less aggregation than without nTHP. At all concentrations tested, SF-THP reduced t(I) (P=0.0001 vs standard conditions) and promoted aggregation (inhibition -48 to -33%); crystals were elongated with a length/width ratio of 4.3+/-0.6 (P<0. 05 vs nTHP). When simultaneously present with nTHP, citrate enhanced the inhibitory effects of nTHP, producing the smallest (length/width 1.5+/-0.1) and least aggregated crystals. Finally, 3.5 mM citrate turned promotory SF-THP into a crystallization inhibitor with abundant small and clustered, but not aggregated crystals. CONCLUSION: Citrate appears to be the main determinant of CaOx crystallization rates and crystal morphology in the presence of nTHP as well as SF-THP. Its effects appear to predominate over those of THP, since even promotory SF-THP is turned into a crystallization inhibitor in the presence of citrate. This re emphasizes at a morphological level what has been concluded from functional as well from clinical studies, namely that citrate is needed in urine at equimolar concentrations to calcium in order to prevent the formation of large crystal aggregates in presence of abnormal THP. PMID- 10692523 TI - Vitamin D metabolite requirements in dialysed children receiving recombinant human growth hormone. AB - BACKGROUND: The aim of the study was to assess the requirement of active vitamin D in dialysed children during treatment with recombinant human growth hormone (rhGH). METHODS: Twenty-six children (aged 5-15 years) were treated with rhGH for 6 months. The serum concentration of parathyroid hormone (PTH), alkaline phosphatase (AP), and calcium and phosphorus were measured in two groups of patients studied in the years 1994-1995 (group I) and 1995-1998 (group II) respectively. Group I received a constant dose of alfacalcidol that was sufficient to keep PTH below 200 pg/ml before rhGH treatment began. The serum PTH level was checked every 3 months. Alfacalcidol was administered to group II according to serum PTH levels checked on a monthly basis. RESULTS: In group I the PTH level increased after 3 and 6 months of rhGH treatment from mean level 73+/ 60; 155+/-156 and 344+/-249 pg/ml respectively; P<0.05. AP activity increased after 6 months of treatment from 206+/-99 to 325+/-124 U/l respectively; P<0.01. The calcium level decreased from baseline after 3 months of treatment from 2.36+/ 0.21 to 2.17+/-0.12 mmol/l respectively; P<0.05. In group II AP activity increased after 3 and 6 months of treatment from 272+/-169 to 332+/-192 and 404. 9+/-219.8 U/l respectively; P<0.01. The mean level of phosphorus decreased after 6 months from 2.15+/-0.28 to 1.70+/-0.39 mmol/l respectively; P<0.01. In group II the mean dose of alfacalcidol increased by 60.9%. CONCLUSIONS: In children with end-stage renal failure, higher doses of vitamin D are needed during rhGH treatment. During rhGH treatment, frequent control of serum PTH level is necessary. PMID- 10692524 TI - Effects of dialyser and dialysate on the acute phase reaction in clinical bicarbonate dialysis. AB - BACKGROUND: In chronic haemodialysis (HD), morbidity may result from repetitive induction of the acute phase response, caused by a bioincompatible dialysis membrane and/or contaminated dialysate. In the present study, cytokine release (interleukin-6, IL-6) and subsequent production of acute phase proteins (C reactive protein, CRP and secretory phospholipase A(2), sPLA(2)) were assessed to investigate whether the HD-induced acute phase reaction depends mainly on the type of membrane or on the sterility of the dialysate. METHODS: In 11 patients, IL-6, CRP and sPLA(2) levels were assessed in blood samples drawn before (t(0)), at the end (t(180)) and 24 h after the start of HD (t(1440)). All patients were dialysed on Cuprammonium (CU) and Polysulphon (PS) dialysers and seven patients underwent an additional HD session on CU plus a dialysate filter (CUf). RESULTS: IL-6 levels were increased significantly at t(180) compared with t(0) (P<0.02) with both CU and CUf. At t(1440), IL-6 levels had returned to baseline. In contrast, marked fluctuations did not occur during HD with PS. At t(180), IL-6 was significantly greater with CU and CUf devices, than with PS (P<0.02). Following HD with CU and CUf, a significant increase in CRP was observed at t(1440), compared with postdialysis values (P/=95% of liver microsomal N dechloroethylation, whereas with IFA as substrate, CYP3A4 catalyzed an average of approximately 70% of liver microsomal N-dechloroethylation (range = 40-90%), with the balance of this activity catalyzed by CYP2B6 (range = 10-70%, dependent on the CYP2B6 content of the liver). Because CYP2B6 can make a significant contribution to human liver microsomal IFA N-dechloroethylation, but only a minor contribution to IFA 4-hydroxylation, the selective inhibition of hepatic CYP2B6 activity in individuals with a high hepatic CYP2B6 content may provide a useful approach to minimize the formation of therapeutically inactive but toxic N dechloroethylated IFA metabolites. PMID- 10692562 TI - Phosphono analogs of glutathione: inhibition of glutathione transferases, metabolic stability, and uptake by cancer cells. AB - Glutathione transferases (GSTs) have been shown to play an important role in multiple drug resistance in cancer chemotherapy. The inactivation of GST isoforms could lead to an enhanced activity of cytotoxic drugs. Thus, we have developed glutathione phosphono analogs [(S)-gamma-glutamyl-(2RS)-(+/-)-2-amino (dialkoxyphosphinyl)-ac etylgl ycines], which were previously shown to be inhibitors of GSTP1-1. In the present study, the inhibition characteristics of these analogs, including isoenzyme specificities, type of inhibition, and determination of K(i) values, were determined. The inhibition of class alpha GSTs was competitive towards GSH. A mixed-type, non-competitive inhibition of class mu and pi GSTs was observed. The K(i) values varied between 880 +/- 210 and 0.45 +/- 0.1 microM. The inhibitors were most effective towards class mu GSTs. In order to investigate the potential use of these GST inhibitors in intact cellular systems, two additional approaches were examined. Firstly, the metabolic stability was tested with purified gamma-glutamyl transpeptidase and cell homogenates as well as during incubation of cell lines. No appreciable degradation was observed in any of the tested systems. Secondly, to facilitate cellular uptake, three derivatives were synthesized in which the glycine carboxylic group was esterified. Uptake and a possible intracellular cleavage to the corresponding free acids were monitored by HPLC analysis. The esters were effectively transported into HT29 (colon cancer) and EPG85-257P (gastric cancer) cells, respectively, and readily converted into the more active free acids. In conclusion, the tested inhibitors may be regarded as model compounds for the development of modulating agents in cancer chemotherapy. PMID- 10692563 TI - Triapine (3-aminopyridine-2-carboxaldehyde- thiosemicarbazone): A potent inhibitor of ribonucleotide reductase activity with broad spectrum antitumor activity. AB - Previous studies from our laboratories have shown that (a) Triapine() is a potent inhibitor of ribonucleotide reductase activity and (b) hydroxyurea-resistant L1210 leukemia cells are fully sensitive to Triapine. In an analogous manner, Triapine was similarly active against the wild-type and a hydroxyurea-resistant subline of the human KB nasopharyngeal carcinoma. Triapine was active in vivo against the L1210 leukemia over a broad range of dosages and was curative for some mice. This agent also caused pronounced inhibition of the growth of the murine M109 lung carcinoma and human A2780 ovarian carcinoma xenografts in mice. Optimum anticancer activity required twice daily dosing due to the duration of inhibition of DNA synthesis which lasted about 10 hr in L1210 cells treated with Triapine in vivo. DNA synthesis in normal mouse tissues (i.e. duodenum and bone marrow) uniformly recovered faster than that in L1210 leukemia cells, demonstrating a pharmacological basis for the therapeutic index of this agent. Triapine was more potent than hydroxyurea in inhibiting DNA synthesis in L1210 cells in vivo, and the effects of Triapine were more pronounced. In addition, the duration of the inhibition of DNA synthesis in leukemia cells from mice treated with Triapine was considerably longer than in those from animals treated with hydroxyurea. Combination of Triapine with various classes of agents that damage DNA (e.g. etoposide, cisplatin, doxorubicin, and 1-acetyl-1,2-bis(methylsulfonyl) 2-(2-chloroethyl)hydrazine) resulted in synergistic inhibition of the L1210 leukemia, producing long-term survivors of tumor-bearing mice treated with several dosage levels of the combinations, whereas no enhancement of survival was found when Triapine was combined with gemcitabine or cytosine arabinoside. The findings demonstrate the superiority of Triapine over hydroxyurea as an anticancer agent and further suggest that prevention by Triapine of repair of DNA lesions created by agents that damage DNA may result in efficacious drug combinations for the treatment of cancer. PMID- 10692564 TI - The relative importance of NADPH: cytochrome c (P450) reductase for determining the sensitivity of human tumour cells to the indolequinone EO9 and related analogues lacking functionality at the C-2 and C-3 positions. AB - Analogues of EO9 (3-hydroxymethyl-5-aziridinyl-1-methyl-2[1H-indole-4-7 dione]prop-2-e n-1-ol) which lack functionality at either the C-2 or C-3 position were synthesised. The aim was to establish the importance of each group towards toxicity and to give an indication as to whether substitution at either position altered activation and toxicity after metabolism by cellular NADPH: cytochrome c (P450) reductase (P450R). MDA231 breast cancer cells were transfected with the cDNA for human P450R and stable clones were isolated. These high P450R-expressing clones were used to determine the aerobic and hypoxic toxicity of EO9 and the two analogues that lacked functionality at either C-2 or C-3. The results showed that P450R was strongly implicated in the bioactivation of EO9 and its analogues under both of these conditions. This data also showed that the C-3 functionality was primarily implicated in hypoxic toxicity. PMID- 10692565 TI - Activation of transcription factors activator protein-1 and nuclear factor-kappaB by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD; dioxin), the prototype agonist of the aromatic hydrocarbon (Ah) receptor, is a potent tumor promoter as well as a complete liver carcinogen that produces an oxidative stress response in rodents and in cultured cell lines. It has been proposed that TCDD promotes neoplastic transformation through oxidative signal transduction pathways, which results in activation of immediate-early response transcription factors. To set the stage for a test of this hypothesis, we evaluated the effect of TCDD treatment on the activation of several transcription factors, including those in the nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) families, which are activated by changes in the redox state of cells. In an extension of prior results, we found that TCDD treatment produced a sustained overexpression of AP-1 for at least 72 hr in wild-type mouse hepatoma Hepa-1 cells, but not in the Ah receptor-deficient derivative c35 or in cytochrome P450-1A1 (CYP1A1)-negative c37 cells. In addition, TCDD treatment caused a significant increase in the DNA binding activity of NF-kappaB, but not in the activities of the other transcription factors tested. AP-1 and NF-kappaB activation were blocked by the thiol antioxidant N-acetylcysteine and by nordihydroguaiaretic acid, an antioxidant and lipooxygenase inhibitor and an inhibitor of the epoxygenase activity of CYP1A1, and did not take place in c35, c37, or in Ah nuclear translator-deficient c4 cells. Hence, sustained activation of these two transcription factors by TCDD is likely to result from a CYP1A1-dependent and Ah receptor complex-dependent oxidative signal. Electrophoretic mobility supershift analyses with specific antibodies showed that most of the increase in NF-kappaB binding activity could be accounted for by increases in p50/p50 complexes. Since these complexes are known to repress NF-kappaB-dependent gene transcription, our results delineate a second molecular mechanism, in addition to the recently found block of tumor necrosis factor-alpha-mediated p50/p65 activation, that may be responsible for the immunosuppresive effects of TCDD. PMID- 10692566 TI - No effect of carvedilol on nitric oxide generation in phagocytes but modulation of production of superoxide ions. AB - Since carvedilol has been claimed to possess antioxidative effects, this drug might affect functional responses, including nitric oxide (NO) generation, of polymorphonuclear neutrophils (PMN) and macrophages. When we assessed the effects of carvedilol on PMN responses in vitro, we observed that carvedilol dose dependently modulated generation of superoxide ions by NADPH oxidase when induced by the formylpeptide formyl-methionyl-leucyl-phenylalanine (fMLP) or the phorbol ester phorbol myristate acetate. This effect was not coupled to diminished phospholipase C activity. In contrast to the effect on NADPH oxidase, neither the fMLP-elicited NO generation by PMN nor the response of the murine macrophage cell line J774 to lipopolysaccharide was affected. There was no evidence from cell free assay systems that carvedilol is a scavenger for superoxide ions or NO. Moreover, carvedilol did not affect other reactions dependent on NO, e.g. spontaneous or fMLP-stimulated PMN migration or lipoxin A(4)-, fMLP-, or A23187 induced neutrophil cytotoxicity for human umbilical vein endothelial cells. Thus, these effects point to the possibility that carvedilol modulates the NADPH oxidase of PMN but leaves the nitric oxide synthase of phagocytes intact. PMID- 10692567 TI - Effect of silymarin on biliary bile salt secretion in the rat. AB - The effect of the hepatoprotector silymarin on bile secretion, with particular regard to bile salt secretion, was studied in Wistar rats. Silymarin (25, 50, 100, and 150 mg/kg/day, i.p., for 5 days) induced a dose-dependent increase in bile flow and bile salt secretion, the maximal effect being reached at a dose of 100 mg/kg/day (+17 and +49%, for bile flow and bile salt output, respectively; P < 0.05). Assessment of bile salt composition in bile revealed that stimulation of the bile salt secretion was accounted for mainly by an increase in the biliary secretion of beta-muricholate and, to a lesser extent, of alpha-muricholate, chenodeoxycholate, ursodeoxycholate, and deoxycholate. The maximum secretory rate (T(m)) of bile salts, as assessed by infusing the non-hepatotoxic bile salt tauroursodeoxycholate i.v. at stepwise-increasing rates, was not influenced by silymarin. The flavonolignan also increased the endogenous bile salt pool size (+53%, P < 0.05) and biliary bile acid excretion after bile acid pool depletion (+54%, P < 0.05), a measure of de novo bile salt synthesis. These results suggest that silymarin increases the biliary excretion and the endogenous pool of bile salts by stimulating the synthesis, among others, of hepatoprotective bile salts, such as beta-muricholate and ursodeoxycholate. PMID- 10692568 TI - Vertebrate pseudogenes. AB - Pseudogenes are commonly encountered during investigation of the genomes of a wide range of life forms. This review concentrates on vertebrate, and in particular mammalian, pseudogenes and describes their origin and subsequent evolution. Consideration is also given to pseudogenes that are transcribed and to the unusual group of genes that exist at the interface between functional genes and non-functional pseudogenes. As the sequences of different genomes are characterised, the recognition and interpretation of pseudogene sequences will become more important and have a greater impact in the field of molecular genetics. PMID- 10692569 TI - Preparation and characterization of 'heparinocytes': erythrocytes with covalently bound low molecular weight heparin. AB - In an attempt to create the possibility of stable, long acting, intravascular anticoagulation, low molecular weight heparin was modified by introducing a sulfhydryl group into the molecule (LMWH-SH). Human erythrocytes were covalently grafted with LMWH-SH by the use of a heterobifunctional coupling reagent which reacts with the SH group of LMWH-SH and surface exposed amino groups of erythrocytes now called 'heparinocytes' (HC). HC were morphologically indistinguishable from untreated erythrocytes and displayed identical osmotic resistance. The functionality of HC was analyzed by classical coagulation tests in which they dose dependently inhibited clot formation. HC were also functional in recalcified whole blood inhibiting thrombin formation as assessed by the cleavage of the chromogenic substrate S-2238. The system appears applicable as a potential autologous, long-term anticoagulant treatment or prophylaxis. PMID- 10692570 TI - Different epitopes are required for gp130 activation by interleukin-6, oncostatin M and leukemia inhibitory factor. AB - Gp130 is the common signal transducing receptor subunit of interleukin (IL)-6, IL 11, leukemia inhibitory factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor and cardiotrophin-1. IL-6 and IL-11 induce gp130 homodimerization whereas the others lead to the formation of heterodimers with LIFR or OSMR. Binding epitopes for IL-6 and IL-11 are located in the immunoglobulin-like domain and the cytokine binding module (CBM). Here we show that a gp130 mutant lacking domain 1, although unresponsive to IL-6 and IL-11, can still activate signal transducer and activator of transcription (STAT) transcription factors in response to LIF or OSM. Moreover, point mutations in the CBM of gp130 (F191E and V252D) that severely impair signal transduction in response to IL-6 and IL-11 differentially interfere with gp130 activation in response to LIF and OSM. Thus, epitopes involved in gp130 homodimerization are distinct from those leading to the formation of gp130/LIFR or gp130/OSMR heterodimers. These findings may serve as the base for rational design of gp130 antagonists that specifically interfere with bioactivity of distinct IL-6-type cytokines. PMID- 10692571 TI - Cytoskeleton alterations of erythrocytes from patients with Fanconi's anemia. AB - Fanconi's anemia (FA) is a very rare genetically heterogeneous disease which has been hypothesized to be defective in the detoxification of reactive oxygen species. In this work we report the results obtained by morphometric and biochemical analyses on the red blood cells (RBCs) from FA patients. With respect to RBCs from healthy donors the following changes have been detected: (i) a variety of ultrastructural alterations, mainly surface blebbing typical of acanthocytes and stomatocytes; (ii) a significant quantitative increase of these altered forms; (iii) modifications of spectrin cytoskeleton network; (iv) an altered redox balance, e.g. a decreased catalase activity and significant variations in the GSSG/GSH ratio. We hypothesize that remodeling of the redox state occurring in FA patients results in cytoskeleton-associated alterations of red blood cell integrity and function. PMID- 10692572 TI - Caspase-induced inactivation of the anti-apoptotic TRAF1 during Fas ligand mediated apoptosis. AB - The activation of the transcription factor NF-kappaB often results in protection against apoptosis. In particular, pro-apoptotic tumor necrosis factor (TNF) signals are blocked by proteins that are induced by NF-kappaB such as TNFR associated factor 1 (TRAF1). Here we show that TRAF1 is cleaved after Asp-163 when cells are induced to undergo apoptosis by Fas ligand (FasL). The C-terminal cleavage product blocks the induction of NF-kappaB by TNF and therefore functions as a dominant negative (DN) form of TRAF1. Our results suggest that the generation of DN-TRAF1 is part of a pro-apoptotic amplification system to assure rapid cell death. PMID- 10692573 TI - Activation of a pro-apoptotic amplification loop through inhibition of NF-kappaB dependent survival signals by caspase-mediated inactivation of RIP. AB - Death domain containing members of the tumor necrosis factor receptor (TNFR) superfamily can induce apoptosis or cell activation. However, the mechanisms by which these opposing programs are selected remain unclear. Frequently, NF-kappaB activation conveys protection against cell death. We show that the serine/threonine kinase RIP that is required for TNF-induced NF-kappaB activation is processed by caspase-8 into a dominant-negative (DN) fragment during death receptor-induced apoptosis, thereby leading to a blockade of NF-kappaB-mediated anti-apoptotic signals. Our results suggest that cleavage of RIP is part of an amplification loop which is triggered by Fas and most likely by other death receptors. PMID- 10692574 TI - Characterization of recombinant mustard trypsin inhibitor 2 (MTI2) expressed in Pichia pastoris. AB - The mustard trypsin inhibitor MTI2 was expressed as secretory protein in the yeast Pichia pastoris. In order to evaluate the influence of the C-terminal amino acids of the precursor form on the inhibitor activity, the C-terminal precursor and the mature protein were both expressed. A third His-tagged construct was also designed to compare alternative purification procedures. Proteins were efficiently expressed at levels of 40-160 mg/l in shake flasks. Equilibrium dissociation constants demonstrated that the mature protein was a stronger inhibitor of bovine beta-trypsin compared to the precursor and His-tagged forms (0.01 nM vs. 0.58 nM and 0.71 nM, respectively). The recombinant proteins were active inhibitors of Spodoptera exigua gut proteases. PMID- 10692575 TI - Two different modes of cyclin clb2 proteolysis during mitosis in Saccharomyces cerevisiae. AB - Sister chromatid separation and mitotic exit are triggered by the anaphase promoting complex (APC/C) which is a multi-subunit ubiquitin ligase required for proteolytic degradation of various target proteins. Cdc20 and Cdh1 are substrate specific activators of the APC/C. It was previously proposed that Cdh1 is essential for proteolysis of the yeast mitotic cyclin Clb2. We show that Clb2 proteolysis is triggered by two different modes during mitosis. A fraction of Clb2 is degraded during anaphase in the absence of Cdh1. However, a second fraction of Clb2 remains stable during anaphase and is degraded in a Cdh1 dependent manner as cells exit from mitosis. Most of cyclin Clb3 is degraded independently of Cdh1. Our data imply that degradation of mitotic cyclins is initiated by a Cdh1-independent mechanism. PMID- 10692576 TI - TFIIA-TAF regulatory interplay: NMR evidence for overlapping binding sites on TBP. AB - TATA box binding protein (TBP)-promoter interaction nucleates assembly of the RNA polymerase II transcription initiation complex. Transcription factor IIA (TFIIA) stabilizes the TBP-promoter complex whereas the N-terminal domain of the largest TAF(II) inhibits TBP-promoter interaction. We have mapped the interaction sites on TBP of Drosophila TAF(II)230 and yeast TFIIA (comprising two subunits, TOA1 and TOA2), using nuclear magnetic resonance (NMR), and also report structural evidence that subdomain II of the TAF(II)230 N-terminal inhibitory domain and TFIIA have overlapping binding sites on the convex surface of TBP. Together with previous mutational and biochemical data, our NMR results indicate that subdomain II augments subdomain I-mediated inhibition of TBP function by blocking TBP-TFIIA interaction. PMID- 10692577 TI - Semi-synthetic Rab proteins as tools for studying intermolecular interactions. AB - Rab GTPases play a key role in the regulation of membrane traffic. Posttranslational geranylgeranylation is critical for their biological activity and is conferred by a Rab geranylgeranyl transferase (RabGGTase). To study the interactions between Rab proteins and RabGGTase, we used in vitro ligation methodology to generate a fluorescent semi-synthetic Rab7 protein. The obtained protein was functionally active and was used to demonstrate a micromolar affinity interaction of Rab7 with the RabGGTase in the absence of Rab escort protein (REP). This finding is consistent with an earlier proposed model according to which RabGGTase possesses two independent weak binding sites for REP and Rab proteins. PMID- 10692578 TI - Olive oil phenolics are dose-dependently absorbed in humans. AB - Olive oil phenolic constituents have been shown, in vitro, to be endowed with potent biological activities including, but not limited to, an antioxidant action. To date, there is no information on the absorption and disposition of such compounds in humans. We report that olive oil phenolics, namely tyrosol and hydroxytyrosol, are dose-dependently absorbed in humans after ingestion and that they are excreted in the urine as glucuronide conjugates. Furthermore, an increase in the dose of phenolics administered increased the proportion of conjugation with glucuronide. PMID- 10692579 TI - Slow formation of [3Fe-4S](1+) clusters in mutant forms of Desulfovibrio africanus ferredoxin III. AB - Desulfovibrio africanus ferredoxin III (Da FdIII) readily interconverts between a 7Fe and an 8Fe form with Asp-14 believed to provide a cluster ligand in the latter form. To investigate the factors important for cluster interconversion in Fe/S cluster-containing proteins we have studied two variants of Da FdIII produced by site-directed mutagenesis, Asp14Glu and Asp14His, with cluster incorporation performed in vitro. Characterisation of these proteins by UV/visible, EPR and (1)H NMR spectroscopies revealed that the formation of the stable 7Fe form of these proteins takes some time to occur. Evidence is presented which indicates the [4Fe-4S](2+) cluster is incorporated prior to the [3Fe 4S](1+) cluster. PMID- 10692580 TI - Dietary flavonoid and isoflavone glycosides are hydrolysed by the lactase site of lactase phlorizin hydrolase. AB - Lactase phlorizin hydrolase (LPH; EC 3.2.1.62) is a membrane-bound, family 1 beta glycosidase found on the brush border of the mammalian small intestine. LPH, purified from sheep small intestine, was capable of hydrolysing a range of flavonol and isoflavone glycosides. The catalytic efficiency (k(cat)/K(m)) for the hydrolysis of quercetin-4'-glucoside, quercetin-3-glucoside, genistein-7 glucoside and daidzein-7-glucoside was 170, 137, 77 and 14 (mM(-1) s(-1)) respectively. The majority of the activity occurred at the lactase and not phlorizin hydrolase site. The ability of LPH to deglycosylate dietary (iso)flavonoid glycosides suggests a possible role for this enzyme in the metabolism of these biologically active compounds. PMID- 10692581 TI - Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein. AB - We previously reported that the nuclear localization signal (NLS) peptides stimulate the in vitro phosphorylation of several proteins, including a 34 kDa protein. In this study, we show that this specific 34 kDa protein is a novel murine leucine-rich acidic nuclear protein (LANP)-like large protein (mLANP-L). mLANP-L was found to have a basic type NLS. The co-injection of Q69LRan-GTP or SV40 T-antigen NLS peptides prevented the nuclear import of mLANP-L. mLANP-L NLS bound preferentially to Rch1 and NPI-1, but not to the Qip1 subfamily of importin alpha. These findings suggest that mLANP-L is transported into the nucleus by Rch1 and/or NPI-1. PMID- 10692582 TI - Human mast cells take up and hydrolyze anandamide under the control of 5 lipoxygenase and do not express cannabinoid receptors. AB - Human mast cells (HMC-1) take up anandamide (arachidonoyl-ethanolamide, AEA) with a saturable process (K(m)=200+/-20 nM, V(max)=25+/-3 pmol min(-1) mg protein( 1)), enhanced two-fold over control by nitric oxide-donors. Internalized AEA was hydrolyzed by a fatty acid amide hydrolase (FAAH), whose activity became measurable only in the presence of 5-lipoxygenase, but not cyclooxygenase, inhibitors. FAAH (K(m)=5.0+/-0.5 microM, V(max)=160+/-15 pmol min(-1) mg protein( 1)) was competitively inhibited by palmitoylethanolamide. HMC-1 cells did not display a functional cannabinoid receptor on their surface and neither AEA nor palmitoylethanolamide affected tryptase release from these cells. PMID- 10692583 TI - A novel rod-like opsin isolated from the extra-retinal photoreceptors of teleost fish. AB - We have isolated a novel opsin from the pineal complex of Atlantic salmon (Salmo salar) and from the brain of the puffer fish (Fugu rubripes). These extra-retinal opsins share approximately 74% identity at the nucleotide and amino acid level with rod-opsins from the retina of these species. By PCR, we have determined that the novel rod-like opsin is not expressed in the salmon retina, and the retinal rod-opsin is not expressed in the salmon pineal. Phylogenetic analysis suggests that the rod-like opsins arose from a gene duplication event approximately 205 million years ago, a time of considerable adaptive radiation of the bony fish. In view of the large differences in the coding sequences of the pineal/brain rod like opsins, their extra-retinal sites of expression, and phylogenetic position we have termed these novel opsins 'extra-retinal rod-like opsins' (ERrod-like opsins). We speculate that the differences between retinal rod-opsins and ERrod like opsins have arisen from their differing photosensory roles and/or genetic drift after the gene duplication event in the Triassic. PMID- 10692584 TI - Identification of lysophospholipid receptors in human platelets: the relation of two agonists, lysophosphatidic acid and sphingosine 1-phosphate. AB - Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (Sph-1-P) are known as structurally related bio-active lipids activating platelets through their respective receptors. Although the receptors for LPA and Sph-1-P have been recently identified in various cells, the identification and characterization of ones in platelets have been reported only preliminarily. In this report, we first investigated the distinct modes of LPA and Sph-1-P actions in platelet activation and found that LPA functioned as a much stronger agonist than Sph-1-P, and high concentrations of Sph-1-P specifically desensitized LPA-induced intracellular Ca(2+) mobilization. In order to identify the responsible receptors underlying these observations, we analyzed the LPA and Sph-1-P receptors which might be expressed in human platelets, by RT-PCR. We found for the first time that Edg2, 4, 6 and 7 mRNA are expressed in human platelets. PMID- 10692585 TI - A novel Drosophila serpin that inhibits serine proteases. AB - Serpins define a large protein family in which most members function as serine protease inhibitors. Here we report the results of a search for serpins in Drosophila melanogaster that are potentially required for oogenesis or embryogenesis. We cloned and sequenced ovarian cDNAs that encode six distinct proteins having extensive sequence similarity to mammalian serpins, including residues important in the serpin inhibition mechanism. One of these new serpins in recombinant form inactivates, and complexes with, trypsin-like proteases in vitro. To our knowledge, these results represent the first evidence for a serpin in Drosophila that functions as a serine protease inhibitor. PMID- 10692586 TI - Origin of the 'inactivation' of ribonuclease A at low salt concentration. AB - The effect of salt concentration on catalysis by ribonuclease A (RNase A) has been reexamined. At low salt concentration, the enzyme is inhibited by low-level contaminants in common buffers. When an uncontaminated buffer system is used or H12A RNase A, an inactive variant, is added to absorb inhibitory contaminants, enzymatic activity is manifested fully at low salt concentration. Catalysis by RNase A does not have an optimal salt concentration. Instead, k(cat)/K(M)10(9) M( 1)s(-1) for RNA cleavage at low salt concentration. These findings highlight the care that must accompany the determination of meaningful salt-rate profiles for enzymatic catalysis. PMID- 10692587 TI - Cloning of a mouse glucocorticoid modulatory element binding protein, a new member of the KDWK family. AB - A mouse cDNA that encodes a nuclear DNA binding protein was identified by yeast two-hybrid screening using the activation domain 2 of the nuclear receptor coactivator TIF2 as a bait. BLAST analysis revealed that the identified cDNA encodes a KDWK domain and contains sequences almost identical to three tryptic peptides of rat GMEB-1 which together with the GMEB-2 heterodimeric partner binds to the GME/CRE sequence (glucocorticoid modulatory element) of the tyrosine aminotransferase (TAT) promoter. Mouse GMEB-1 is ubiquitously expressed in all the tissues examined. In vitro translated mGMEB-1 bound specifically to GME oligonucleotides, either alone or as a heterodimer with rGMEB-2. Transient transfection experiments with TAT promoter reporter genes suggest a potential role for mGMEB-1 as a transcriptional regulator of the TAT promoter. PMID- 10692588 TI - Radiation inactivation analysis of H(+)-pyrophosphatase from submitochondrial particles of etiolated mung bean seedlings. AB - Radiation inactivation analysis was employed to determine the functional masses of enzymatic activity and proton translocation of H(+)-pyrophosphatase from submitochondrial particles of etiolated mung bean seedlings. The activities of H(+)-pyrophosphatase decayed as a simple exponential function with respect to radiation dosage. D(37) values of 6.9+/-0.3 and 7.5+/-0.5 Mrad were obtained for pyrophosphate hydrolysis and its associated proton translocation, yielding molecular masses of 170+/-7 and 156+/-11 kDa, respectively. In the presence of valinomycin and 50 mM KCl, the functional size of H(+)-pyrophosphatase of tonoplast was decreased, while that of submitochondrial particles remained the same, indicating that they are two distinct types of proton pump using PP(i) as their energy source. PMID- 10692589 TI - Isolation of recombinant BMP receptor IA ectodomain and its 2:1 complex with BMP 2. AB - Bone morphogenetic protein-2 (BMP-2) is a member of the transforming growth factor beta superfamily which induces bone formation and regeneration, and important steps during early embryonic development. BMP-2 signals via oligomerization of type I and type II serine/threonine kinase receptors. We report here expression of the extracellular domain of the human type IA receptor for BMP-2 (BMPR-IA) in Escherichia coli. This soluble form of BMPR-IA (sBMPR-IA) was purified employing a BMP-2 affinity column. Gel filtration experiments and analysis of gel filtration fractions by polyacrylamide electrophoresis and densitometry reveal that BMP-2 forms a defined 1:2 complex with sBMPR-IA that can be purified and hopefully used for crystallization studies. PMID- 10692590 TI - Identification of active site serine and histidine residues in Escherichia coli outer membrane protease OmpT. AB - Escherichia coli outer membrane protease OmpT has been characterised as a serine protease based on its inhibitor profile, but serine protease consensus sequences are absent. By site-directed mutagenesis we substituted all conserved serines and histidines. Substitution of His(101) and His(212) by Ala, Asn or Gln resulted in variant enzymes with 0.01 and 9-20% residual enzymatic activity towards a fluorogenic pentapeptide substrate, respectively. The mutations S140A and S201A did not decrease activity, while variants S40A and S99A yielded 0.5 and 0.2% residual activities, respectively. When measured with a dipeptide substrate the variant S40A demonstrated full activity, whereas variant S99A displayed at least 500-fold reduced activity. We conclude that Ser(99) and His(212) are essential active site residues. We propose that OmpT is a novel serine protease with Ser(99) as the active site nucleophile and His(212) as general base. PMID- 10692591 TI - The C-terminal tetrapeptide HWFW of the Chlorella HUP1 hexose/H(+)-symporter is essential for full activity and an alpha-helical structure of the C-terminus. AB - C-terminal tails of plant hexose/H(+)-symporters of the major facilitator superfamily contain a highly conserved motif of four amino acids: HWFW. A deletion of these four amino acids in the Chlorella HUP1 protein leads to a decrease in transport activity by a factor of 3-4. The mutated tail is highly sensitive to trypsin; it does not show alpha-helical conformation in contrast to the wild type C-terminal peptide with an alpha-helical content of at least 15%. The production of monoclonal antibody 416B8 recognizing an epitope within the central loop of HUP1 protein has been a prerequisite for the experiments described. PMID- 10692592 TI - A novel covalent modification of nitrogenase in a cyanobacterium. AB - In extracts of the unicellular cyanobacterium Gloeothece, the Fe-protein of nitrogenase can be separated by SDS-PAGE into two antigenically identifiable components. Unlike the situation in photosynthetic bacteria such as Rhodospirillum rubrum, these two forms do not arise from covalent modification of the protein by ADP-ribosylation. Rather, the Fe-protein of Gloeothece nitrogenase is subjected to modification by palmitoylation. PMID- 10692593 TI - Deficiency of a STE20/PAK family kinase LOK leads to the acceleration of LFA-1 clustering and cell adhesion of activated lymphocytes. AB - Lymphocyte-oriented kinase (LOK) is a member of the STE20/p21-activated kinase (PAK) family and expressed predominantly in lymphoid organs. Generation of LOK deficient mice revealed that the leukocyte-function-associated antigen (LFA 1)/intercellular adhesion molecules (ICAM)-mediated aggregation of mitogen stimulated T cells was greatly enhanced in the absence of LOK. Though levels of total LFA-1 and ICAMs as well as the active form of LFA-1 on T cell blasts were comparable in the presence and absence of LOK, clustering of active LFA-1 detected by binding of soluble ICAM-1 was accelerated in the absence of LOK. These results suggest that LOK is potentially involved in the regulation of LFA-1 mediated lymphocyte adhesion. PMID- 10692594 TI - Chronic ethanol ingestion increases efficiency of oxidative phosphorylation in rat liver mitochondria. AB - The efficiency of oxidative phosphorylation was compared between rats chronically fed with ethanol and controls. (i) Results showed that the liver mitochondria state 4 respiratory rate was strongly inhibited, while the corresponding proton motive force was not affected; (ii) the cytochrome oxidase content and activity were decreased and (iii) the oxidative-phosphorylation yield was increased in the ethanol exposed group. Furthermore, oxidative phosphorylation at coupling site II was not affected by ethanol. Cytochrome oxidase inhibition by sodium-azide mimicked the effects of ethanol intoxication in control mitochondria. This indicates that the decrease in cytochrome oxidase activity induced by ethanol intoxication directly increases the efficiency of oxidative phosphorylation. PMID- 10692595 TI - Purification and characterisation of epithiospecifier protein from Brassica napus: enzymic intramolecular sulphur addition within alkenyl thiohydroximates derived from alkenyl glucosinolate hydrolysis. AB - Epithiospecifier protein (ESP), a ferrous ion dependent protein, has a potential role in regulating the release of elemental sulphur, nitriles, isothiocyanates and cyanoepithioalkanes from glucosinolates. Two classes of ESP polypeptides were purified with molecular masses of 39 and 35 kDa, and we show that the previously reported instability was conditionally dependent. The 39 kDa polypeptide was made up of two distinct isozymes (5.00, 5.14) whilst several were present for the 35 kDa form of ESP (5.40-5.66). An anti-ESP antibody reacted with both the 39 and 35 kDa ESP forms in Brassica napus and strongly with a polypeptide corresponding to the 35 kDa ESP form in Crambe abyssinica, but did not detect any ESP in Sinapis alba or Raphanus sativus. A cytochrome P-450 mediated iron dependent epoxidation type mechanism is suggested for ESP. PMID- 10692596 TI - Clinical notes PMID- 10692597 TI - Trigeminal neuralgia: opportunities for research and treatment. AB - Trigeminal neuralgia was the focus of a recent workshop convened by the National Institute of Neurological Disorders and Stroke (NINDS) and the National Institute of Dental and Craniofacial Research (NIDCR). The workshop brought together basic scientists, clinicians, epidemiologists, and patient advocates. New research directions for epidemiology, diagnosis and assessment, pain mechanisms, and treatment were identified. (The workshop was held in Rockville MD on September 14, 1999, with financial support from NINDS, NIDCR, the NIH Office of Rare Diseases, and the NIH Pain Research Consortium.) PMID- 10692598 TI - Effects of antagonists to high-threshold calcium channels upon spinal mechanisms of pain, hyperalgesia and allodynia. AB - High-threshold voltage-dependent calcium channels enable calcium ions to enter neurons upon depolarization and thereby influence synaptic mediator/receptor systems, membrane excitability levels, second and third messenger concentration, and gene expression. These phenomena underlie several processes including those of normal nociception and of hyperalgesia and allodynia. The present article deals with the role of spinal L-, N- and P/Q-type calcium channels in short lasting nociception as well as in the hyperalgesia and allodynia elicited by chemical irritants of peripheral nociceptors, inflammatory and mechanical lesions of peripheral tissues, and lesions of peripheral nerves. The studies summarized herein are based on the spinal delivery of specific antagonists to high-threshold calcium channels, and reveal that blockade of L-type, P/Q-type and, particularly, N-type channels can prevent, attenuate, or both, subjective pain as well as primary and/or secondary hyperalgesia and allodynia in a variety of experimental and clinical conditions. PMID- 10692599 TI - Pain-related cerebral activation is altered by a distracting cognitive task. AB - It has previously been suggested that the activity in sensory regions of the brain can be modulated by attentional mechanisms during parallel cognitive processing. To investigate whether such attention-related modulations are present in the processing of pain, the regional cerebral blood flow was measured using [(15)O]butanol and positron emission tomography in conditions involving both pain and parallel cognitive demands. The painful stimulus consisted of the standard cold pressor test and the cognitive task was a computerised perceptual maze test. The activations during the maze test reproduced findings in previous studies of the same cognitive task. The cold pressor test evoked significant activity in the contralateral S1, and bilaterally in the somatosensory association areas (including S2), the ACC and the mid-insula. The activity in the somatosensory association areas and periaqueductal gray/midbrain were significantly modified, i.e. relatively decreased, when the subjects also were performing the maze task. The altered activity was accompanied with significantly lower ratings of pain during the cognitive task. In contrast, lateral orbitofrontal regions showed a relative increase of activity during pain combined with the maze task as compared to only pain, which suggests the possibility of the involvement of frontal cortex in modulation of regions processing pain. PMID- 10692600 TI - Pain and allodynia/hyperalgesia induced by intramuscular injection of serotonin in patients with fibromyalgia and healthy individuals. AB - The aim of this study was to investigate the effect of injection of serotonin (5 HT) into the masseter muscle on pain and allodynia/hyperalgesia. Twelve female patients with fibromyalgia (FM) and 12 age-matched female healthy individuals (HI) participated in the study. The current pain intensity (CPI) and the pressure pain threshold (PPT) of the superficial masseter muscles were assessed bilaterally. 5-HT in one of three randomized concentrations (10(-3), 10(-5), 10( 7) M) or isotonic saline was then injected into either of the two masseter muscles in a double-blind manner. After the injections the CPI and PPT were recorded ten times during 30 min. The injections were repeated twice with the other concentrations of 5-HT after 1 and 2 weeks, respectively. In the FM-group there was a non-significant increase of CPI after injection that lasted during the entire 30-min period irrespective of whether 5-HT or saline was injected. Neither did the PPT change significantly. In the HI-group pain developed significantly after injection irrespective of whether 5-HT or saline was injected, but significantly more so after 5-HT at 10(-3) M than saline injection. CPI decreased quickly and then remained on a very low level for most of the experiment. 5-HT at both 10(-5) M and 10(-3) M caused a significantly greater decrease of PPT than saline. In conclusion, our results show that 5-HT injected into the masseter muscle of healthy female subjects elicits pain and allodynia/hyperalgesia, while no such responses occur in patients with fibromyalgia. PMID- 10692601 TI - Immunolocalization of SNS/PN3 and NaN/SNS2 sodium channels in human pain states. AB - The tetrodotoxin-resistant (TTX-R) voltage-gated sodium channel SNS/PN3 and the newly discovered NaN/SNS2 are expressed in sensory neurones, particularly in nociceptors. Using specific antibodies, we have studied, for the first time in humans, the presence of SNS/PN3 and NaN/SNS2 in peripheral nerves, including tissues from patients with chronic neurogenic pain. In brachial plexus injury patients, there was an acute decrease of SNS/PN3- and NaN/SNS2-like immunoreactivity in sensory cell bodies of cervical dorsal root ganglia (DRG) whose central axons had been avulsed from spinal cord, with gradual return of the immunoreactivity to control levels over months. In contrast, there was increased intensity of immunoreactivity to both channels in some peripheral nerve fibers just proximal to the site of injury in brachial plexus trunks, and in neuromas. These findings suggest that the expression of these sodium channels in neuronal cell bodies is reduced after spinal cord root avulsion injury in man, but that pre-synthesized channel proteins may undergo translocation with accumulation at sites of nerve injury, as in animal models of peripheral axotomy. The latter may contribute to positive symptoms, as our patients all showed a positive Tinel's sign. Nerve terminals in distal limb neuromas and skin from patients with chronic local hyperalgesia and allodynia all showed marked increases of SNS/PN3 immunoreactive fibers, but little or no NaN/SNS2-immunoreactivity, suggesting that the former may be related to the persistent hypersensitive state. Axonal immunoreactivity to both channels was similar to control nerves in sural nerve biopsies in a selection of neuropathies, irrespective of nerve inflammation, demyelination or spontaneous pain, including a patient with congenital insensitivity to pain. Our studies suggest that the best target for SNS/PN3 blocking agents is likely to be chronic local hypersensitivity. PMID- 10692602 TI - Age-related differences in the time course of capsaicin-induced hyperalgesia. AB - The effect of age on hyperalgesia, one of the most common signs of injury, has not been previously examined in humans. A psychophysical study was conducted in 10 young (26.9+/-4.6 years) and 10 older (79. 0+/-5.7 years) healthy volunteers to investigate the effect of age on the development of hyperalgesia induced by topical application of capsaicin (0.1 ml, 5 mg/ml). The capsaicin patch (diameter 2 cm) was applied for 1 h. The intensity of capsaicin-induced spontaneous sensation, mechanical pain threshold, area of flare, heat and punctate hyperalgesia were measured hourly for 3 h after the application. Older adults took a longer period to report first pain. There was no age effect on the magnitude of spontaneous sensation, flare size and area of heat hyperalgesia. The area of heat hyperalgesia rapidly decreased over time in both age groups. In marked contrast, the area of punctate hyperalgesia and associated reduction in the mechanical pain threshold were maintained in older adults over the entire 3 h test period, but resolved rapidly in young adults. We conclude that, given the same intensity of noxious stimulation, older adults display a similar magnitude of hyperalgesia as younger persons. However, once initiated, punctate hyperalgesia appears to resolve more slowly in older people. This finding may indicate age differences in the plasticity of spinal cord neurons following an acute injury. PMID- 10692603 TI - Intrathecal lithium reduces neuropathic pain responses in a rat model of peripheral neuropathy. AB - We tested the ability of lithium (Li(+)) to block heat hyperalgesia, cold allodynia, mechanical allodynia and mechanical hyperalgesia in rats experimentally subjected to painful peripheral neuropathy. Chronic constrictive injury (CCI) to the sciatic nerve induced persistent hyperalgesia and allodynia. Intrathecal injection of Li(+) (2.5-40 micromol) into the region of lumbar enlargement dose-dependently reduced heat hyperalgesia, cold allodynia and mechanical allodynia for 2-6 h after injection, but had no effect on mechanical hyperalgesia. Li(+) had no significant effect on responses from control and sham operated animals. Intrathecal injection of myo-inositol (2.5 mg) significantly reversed both the anti-hyperalgesic and anti-allodynic effect of Li(+). These findings suggest that intrathecal Li(+) suppresses neuropathic pain response in CCI rats through the intracellular phosphatidylinositol (PI) second messenger system in spinal cord neurons. Lithium (Li(+)) has already found widespread clinical application; these results suggest that its therapeutic utility may be extended to include treatment of neuropathic pain syndromes resulting from peripheral nerve injury. PMID- 10692604 TI - Morphine and NMDA receptor antagonism reduce c-fos expression in spinal trigeminal nucleus produced by acute injury to the TMJ region. AB - Pain management in temporomandibular disorders (TMDs) often involves pharmacotherapy; however, the site of action for drugs that reduce TMD pain is not known. To determine possible central neural targets of analgesic drugs relevant in TMD pain, morphine or the N-methyl-D-aspartate receptor antagonist, MK-801, was given alone or in combination prior to TMJ injury. The number of neurons expressing the immediate early gene, c-fos, was quantified in the lower brainstem and upper cervical spinal cord as an index of neural activation. It was hypothesized that those neuronal groups most necessary for the sensory discriminative aspects of acute TMJ injury should display the greatest reduction in c-fos expression after drug treatment. Barbiturate-anesthetized male rats were given morphine or MK-801 15 min prior to injection of mustard oil into the TMJ region. Morphine given centrally (i.c.v.) or peripherally (i.v.) caused a marked dose-related reduction in Fos-like immunoreactivity (Fos-LI) in laminae I-II at the middle portions of subnucleus caudalis (mid-Vc) and at the subnucleus caudalis/upper cervical spinal cord (Vc/C2) transition. Higher doses of morphine also reduced Fos-LI in the dorsal paratrigeminal region (dPa5) and at the subnucleus interpolaris/subnucleus caudalis (Vi/Vc-vl) transition. MK-801 given i.v. reduced Fos-LI only in laminae I-II at the Vc/C2 transition. Combined subthreshold doses of morphine and MK-801 reduced c-fos expression in the dPa5, mid-Vc, and the Vc/C2 transition region, below that predicted from the effects of either drug alone. These results suggest that neurons in laminae I-II of the mid Vc and Vc/C2 transition and, to a lesser extent, in the dPa5 region play a critical role in mediating the sensory and/or reflex aspects of pain after acute injury to the TMJ region. PMID- 10692605 TI - Differences between the sexes in post-surgical pain. AB - It has been shown that women have a lower pain threshold and lower tolerance to some forms of experimental pain then men. However, the evidence that clinical pain is perceived differently by the two sexes is not yet as strong. The placement of intraoral implants is a highly controlled surgical procedure that we have used to investigate this possibility. Forty-eight edentulous (without teeth) subjects (27 females), aged from 35 to 63 years, received two titanium implants in the anterior mandible under local anesthesia. After the surgery, subjects completed a pain diary three times each day, rating pain intensity and unpleasantness on 100 mm visual analog scales (VAS). Once a day, they chose verbal descriptors from the McGill Pain Questionnaire (MPQ). Age of subjects, duration of surgery, the amount of local anesthetic used and the amount of pain medication taken were not statistically different for the two groups (P>/=0.32). Results showed that the senior surgeon produced significantly less pain than a 4th year resident (P=0.04). Although there were no significant differences between sexes for mean daily ratings of intensity or unpleasantness over time (P>/=0.10), most women experienced the highest intensity of pain during the day, while most men had higher pain in the evening (P=0.025). Also, the relative unpleasantness (unpleasantness/intensity ratio) increased significantly with time for males, but not for females (P=0.016). Males and females did not differ in the total number of words chosen from the MPQ (P=0.61), or in the averaged Pain Rating Index (PRI) (P=0.53). However, women used significantly more evaluative words than men (P=0.04), suggesting that woman found the overall intensity greater. These results indicate that women find post-surgical pain more intense than males, but that men are more disturbed than women by low levels of pain that last several days. PMID- 10692606 TI - Sociodemographic correlates of tooth pain among adults: United states, 1989. AB - This study presents the sociodemographic distribution of tooth pain and the dental care utilization of affected individuals. Data for adults 20 years of age and over were derived from the 1989 National Health Interview Survey's supplements on dental health, orofacial pain, and health insurance (n=33073). Prevalence of tooth pain by socioeconomic status (SES) and adjusted odds ratios of reporting tooth pain in the past 6 months and of having no dental visits in the past year among persons reporting pain in the previous 6 months were computed taking into account the survey's complex sample design. Tooth pain in the past 6 months was reported by 14.5% (95% CI 14.0, 15.0) of adults aged 20-64 years and by 7.0% (95% CI 6.1, 7.9) of those 65 years and over. In the younger age group, tooth pain was more likely to be reported by those with low SES than it was by those with high SES; in the older age group, tooth pain was more likely reported by non-Hispanic blacks than it was by non-Hispanic whites or Hispanics. Of those reporting pain, younger and older non-Hispanic blacks and persons with lower educational attainment were more likely not to have a dental visit in the previous 12 months. Persons with low SES characteristics were more likely to report tooth pain and to endure their pain without the benefit of dental care while the pain was present. PMID- 10692607 TI - Sex differences and phases of the estrous cycle alter the response of spinal cord dynorphin neurons to peripheral inflammation and hyperalgesia. AB - The neuromodulatory interactions of sex steroids with the opioid system may result in sex differences in pain and analgesia. Dynorphin is an endogenous kappa opioid peptide that is upregulated in an animal model of peripheral inflammation and hyperalgesia and is possibly regulated by circulating levels of sex steroids. The present study compared behavioral responses of male, cycling female, and gonadectomized Sprague-Dawley rats in a model of persistent pain. Cycling female rats were behaviorally tested over a 14-day period, and their estrous cycles were monitored by daily vaginal smears. Thermal hyperalgesia was measured by paw withdrawal latencies taken prior to and 24-72 h after rats received a unilateral hindpaw injection of complete Freund's adjuvant (CFA). Prior to CFA administration, there was no significant difference in paw withdrawal latencies between male rats, cycling female rats, and ovariectomized female rats. Following CFA administration, female rats in proestrus exhibited significantly increased hyperalgesia compared with male rats, ovariectomized female rats, and female rats in other estrous stages (P/=2 months) use of NSAIDs which gave information on gastroduodenal ulcer, bleed or perforation, death due to these complications, or progression from one level of harm to the next. Fifteen RCTs (19364 patients exposed to NSAIDs for 2-60 months), three cohort studies (215076 patients redeeming a NSAID prescription over a 3-12 month period), six case-control studies (2957 cases) and 20 case series (7406), and case reports (4447) were analysed. In RCTs the incidence of bleeding or perforation in 6822 patients exposed to NSAIDs was 0.69%; two deaths occurred. Of 11040 patients with bleeding or perforation with or without NSAID exposure across all reports, 6-16% (average 12%) died; the risk was lowest in RCTs and highest in case reports. Death from bleeding or perforation in all controls not exposed to NSAIDs occurred in 18 out of 849489 (0.002%). From these numbers we calculated the number-needed-to-treat for one patient to die due to gastroduodenal complications with chronic (>/=2 months) NSAIDs as 1/((0.69x?6-16%, average 12%?) 0.002%))=909-2500 (average 1220). On average 1 in 1200 patients taking NSAIDs for at least 2 months will die from gastroduodenal complications who would not have died had they not taken NSAIDs. This extrapolates to about 2000 deaths each year in the UK. PMID- 10692615 TI - Effects of the I(1) imidazoline/alpha(2)-adrenergic receptor agonist moxonidine in comparison with clonidine in the formalin test in rats. AB - Moxonidine is a mixed I(1) imidazoline/alpha(2)moxonidine=morphine. The I(1) imidazoline preferring antagonist efaroxan produced a dose-dependent antagonism of both moxonidine (5.0 mg/kg) and clonidine (0.5 mg/kg). In addition, the alpha(2)-adrenergic receptor antagonist yohimbine produced a dose-related antagonism of moxonidine, but only partially antagonized clonidine. Prazosin failed to block the effects of either moxonidine or clonidine, indicating a lack of involvement of alpha(1) as well as alpha(2B) and alpha(2C) receptors. The present results suggest that alpha(2)-adrenergic receptors play an important role in mediating the effects of moxonidine in producing antinociception in the formalin test. Further, the present results demonstrate that the mechanism of action of moxonidine and clonidine differ in that clonidine, but not moxonidine, produces an antinociceptive effect through a yohimbine-insensitive mechanism in the formalin test. PMID- 10692617 TI - Periaqueductal gray matter metabotropic glutamate receptors modulate formalin induced nociception. AB - The role played by periaqueductal gray (PAG) matter metabotropic glutamate receptors (mGluRs) in the modulation of persistent noxious stimulation was investigated in mice. The formalin test was used as a model of persistent pain. Intra-PAG microinjections of (S)-3, 5-DHPG (25 and 50 nmol/mouse) and L-CCG-I (30 and 60 nmol/mouse), agonists of group I and group II mGluRs, respectively, decreased the nociceptive response (-92+/-6% and -89+/-8%, respectively) during the late phase. No change of the early nociceptive phase was observed after (S) 3,5-DHPG or L-CCG-I treatments. These effects were antagonized by a pretreatment with CPCCOEt (40 nmol/mouse) and (2S)-alpha-EGlu (30 nmol/mouse). CPCCOEt is a selective antagonist of group I mGlu receptors, while (2S)-alpha-EGlu is an antagonist of group II. Intra-PAG microinjections of L-SOP (60 and 120 nmol/mouse), a selective agonist of group III mGluRs, induced an increase of the nociceptive response (+95+/-7%) during the late hyperalgesic phase. (R,S)-alpha-M SOP (70 nmol/mouse), a selective antagonist of group III mGluRs, completely antagonized the L-SOP-induced effect. These results show that PAG mGluRs participate in modulating the late hyperalgesic behaviours induced by formalin. It seems, therefore, possible that group I and group II mGluRs positively modulate PAG antinociceptive descending pathway following a persistent noxious stimulation, while group III mGluRs modulate it negatively. PMID- 10692618 TI - The impact of HIV infection on primary headache. Unexpected findings from retrospective, cross-sectional, and prospective analyses. AB - Headache is one of the most important factors influencing the quality of life in patients infected with the human immunodeficiency virus type 1 (HIV). However, only symptomatic headache but not changes or primary headache types during HIV infection have been studied to date. Therefore, we aimed to determine the impact of an HIV infection on frequency and semiology of different primary headache types. Patients with confirmed HIV type 1 infection underwent a neurological examination, neuroimaging or EEG, and a standardized interview. Time pattern and symptoms of headaches (cross-sectional analysis), changes of headaches preexisting to their infection (longitudinal retrospective analysis), and changes of primary headaches during a 2-year follow-up (longitudinal prospective analysis) were evaluated as were the correlations between these headache patterns and different markers of HIV infection. One hundred thirty-one consecutive HIV infected patients without evidence of a cerebral manifestation except mild encephalopathy were enrolled. The point prevalence of migraine was 16.0% (confidence interval (CI) 10.1-25.4%), of headache with a semiology of tension type headache 45.8% (CI 33.7-62.2%), and of other headache types 6.1% (CI 3.0 12.5%). During the natural course of infection, the migraine frequency significantly decreased in the retrospective and in the prospective analyses, whereas the frequency of the headache with a semiology of tension-type headache significantly increased in all three analyses. In 20% of all patients, the tension-type headache could be considered as symptomatic due to the infection but not due to focal or general cerebral lesions. Changes of primary headache were significantly associated with different stages of the infection and with the presence of mild encephalopathy but not with antiretroviral treatment or CD4 cell count. HIV infection seems to be associated with a progressive decrease in migraine frequency and intensity which probably is related to the immunological state of the patients. Tension-type headache becomes more frequent during HIV infection. However, this can in part be related to secondary headache caused by the HIV in less than 50% of patients with tension-type headache. The progressing immunological deficiency of HIV-infected patients seems to influence pain processing of primary headache types in different ways. PMID- 10692619 TI - Perceived pain before and after three exercise programs--a controlled clinical trial of women with work-related trapezius myalgia. AB - The effect of exercise on neck-shoulder pain was studied in 103 women with work related trapezius myalgia randomized into three exercise groups and a control group. One group trained strength, the second muscular endurance and the third co ordination. The exercise groups met three times weekly for 10 weeks. Pain assessment was made on three visual analogue scales, indicating pain at present, pain in general and pain at worst. Pain thresholds were measured in the trapezius muscle with a pressure algometer. A pain drawing was completed. The rated pain decreased significantly (P<0.05) on the VAS describing pain at worst in the strength and endurance groups. Pressure sensitivity decreased significantly (P<0.05) in four triggerpoints in the exercise groups. No changes were seen in the extent of painful body area in any group. Comparison of exercisers (n=82) and controls (n=21) showed significantly larger pain reductions on VAS pain at present and VAS pain at worst among exercisers. All three exercise programs showed similar decreases of pain which indicates that the type of exercise is of less importance to achieve pain reduction. PMID- 10692620 TI - The cognitive and psychomotor effects of morphine in healthy subjects: a randomized controlled trial of repeated (four) oral doses of dextropropoxyphene, morphine, lorazepam and placebo. AB - Ten healthy subjects (four male) of mean age 31 years (range 25-40) took part in a randomized double-blind four-way crossover study to examine the cognitive and psychomotor effects of repeated oral doses of dextropropoxyphene and morphine. Four treatments were compared: dextropropoxyphene napsylate 100 mg, morphine sulphate 10 mg, lorazepam 0.5 mg and placebo. Four doses of each drug were given at 4-h intervals to each subject on four separate study days at least 1 week apart. Cognitive function was assessed using choice reaction time, number vigilance, memory scanning, immediate and delayed word recall, word recognition, picture recognition, critical flicker fusion threshold (CFFT) and subjective measures of alertness, calmness and contentment. Lorazepam impaired the speed of responding on all tasks in which speed was recorded (except digit vigilance) and increased subjective ratings of calmness. Morphine had one major effect, which was to increase the accuracy of responding on the choice reaction time task, at every assessment. Morphine produced some sporadic effects in other tests and an increase in subjective calmness. Dextropropoxyphene impaired performance on choice reaction time and picture recognition. These data show that oral morphine may enhance performance in some measures of cognitive function, whereas dextropropoxyphene (in usual therapeutic doses) seems more likely to cause impairment. Neither opioid has substantial effects on cognition and psychomotor function compared with lorazepam. PMID- 10692621 TI - Low-dose lidocaine reduces secondary hyperalgesia by a central mode of action. AB - Sodium channel blockers are approved for intravenous administration in the treatment of neuropathic pain states. Preclinical studies have suggested antihyperalgesic effects on the peripheral as well as the central nervous system. The objective of this study was to determine mechanisms of action of low-dose lidocaine in experimental induced, secondary hyperalgesia. In a first experimental trial, participants (n=12) received lidocaine systemically (a bolus injection of 2 mg/kg in 10 min followed by an intravenous infusion of 2 mg kg( 1)h(-1) for another 50 min). In a second trial, a modified intravenous regional anesthesia (IVRA) was administered to exclude possible central analgesic effects. In one arm, patients received an infusion of 40 ml lidocaine, 0.05%; in the other arm 40 ml NaCl, 0.9%, served as a control. In both trials capsaicin, 20 microgram, was injected intradermally and time course of capsaicin-induced pain, allodynia and hyperalgesia as well as axon reflex flare was determined. The capsaicin-induced pain was slightly reduced after systemic and regional application of the anesthetic. The area of pin-prick hyperalgesia was significantly reduced by systemic lidocaine, whereas the inhibition of hyperalgesia was absent during regional administration of lidocaine. In contrast, capsaicin-induced flare was significantly decreased after both treatments. We conclude that systemic lidocaine reduces pin-prick hyperalgesia by a central mode of action, which could involve blockade of terminal branches of nociceptors. A possible role for tetrodotoxin resistant sodium channels in the antihyperalgesic effect of low-dose lidocaine is discussed. PMID- 10692622 TI - Avoidance versus focused attention and the perception of pain: differential effects for men and women. AB - The aim of the current investigation was to compare the effects of two different attentional strategies (focused vs. avoidance) on how males and females respond to experimentally induced pain. One hundred healthy adults were instructed to either attend towards or away from cold pressor pain. Measures of pain tolerance, pain threshold and recovery were taken, as were self-report measures of sensory and affective pain experiences. As expected, gender was found to moderate tolerance to pain: males were found to be more tolerant to cold pressor pain than females. With respect to the self-report measures, males reported less sensory pain when they attended toward the pain than when they avoided it. However, a similar effect was not found in women, suggesting that attentional focusing may only be a useful strategy for males. These results are discussed in light of previous research. PMID- 10692624 TI - The role of 5-HT(1A)-receptors in fentanyl-induced bulbospinal inhibition of a spinal withdrawal reflex in the rabbit. AB - The sural to gastrocnemius withdrawal reflex is inhibited after injection of the OP(3) (micro)-receptor-selective opioid fentanyl into the fourth ventricle of decerebrated rabbits. This effect is abolished by complete section of the spinal cord but not by the selective alpha(2)-adrenoceptor antagonist RX 821002 (Clarke RW, Parry-Baggott C, Houghton AK, Ogilvie J. The involvement of bulbo-spinal pathways in fentanyl-induced inhibition of spinal withdrawal reflexes in the decerebrated rabbit. Pain 1998;78:197-207). We have now investigated the role of 5-HT(1A) receptors in mediating the descending inhibition activated by intraventricular fentanyl. In the control state, intraventricular fentanyl (3-30 microgram/kg) inhibited gastrocnemius reflex responses to a median of 34% of pre drug levels. After intrathecal administration of the selective 5-HT(1A) receptor antagonist WAY-100635 (100 microgram), fentanyl reduced reflex responses to 83% of pre-fentanyl values, significantly less inhibition than in the control state. In a separate group of experiments, intravenous fentanyl (0.3-30 microgram/kg) depressed the sural-gastrocnemius reflex to 17% of pre-drug controls. This inhibition was not affected by intrathecal WAY-100635 (100 microgram), but combined administration of the 5-HT(1A) antagonist with RX 821002 (100 microgram) significantly reduced the effectiveness of i.v. fentanyl. After the highest dose reflexes were 37% of pre-fentanyl levels. These data show that the bulbospinal inhibition activated by fentanyl is mediated, at least in part, by activation of spinal 5-HT(1A) receptors. That blockade of these receptors failed to influence the inhibition induced by i.v. fentanyl might be taken to mean that the brain stem action of fentanyl does not contribute significantly to the systemic actions of this opioid. A more probable explanation is that, in the preparation used in the present study, the bulbospinal and direct spinal actions of fentanyl occlude each other to produce an overall inhibition that is less than the sum of the two effects. PMID- 10692623 TI - Do nerve growth factor-related mechanisms contribute to loss of cutaneous nociception in leprosy? AB - While sensory loss in leprosy skin is the consequence of invasion by M. leprae of Schwann cells related to unmyelinated fibres, early loss of cutaneous pain sensation, even in the presence of nerve fibres and inflammation, is a hallmark of leprosy, and requires explanation. In normal skin, nerve growth factor (NGF) is produced by basal keratinocytes, and acts via its high affinity receptor (trk A) on nociceptor nerve fibres to increase their sensitivity, particularly in inflammation. We have therefore studied NGF- and trk A-like immunoreactivity in affected skin and mirror-site clinically-unaffected skin from patients with leprosy, and compared these with non-leprosy, control skin, following quantitative sensory testing at each site. Sensory tests were within normal limits in clinically-unaffected leprosy skin, but markedly abnormal in affected skin. Sub-epidermal PGP 9.5- and trk A- positive nerve fibres were reduced only in affected leprosy skin, with fewer fibres contacting keratinocytes. However, NGF-immunoreactivity in basal keratinocytes, and intra-epidermal PGP 9.5-positive nerve fibres, were reduced in both sites compared to non-leprosy controls, as were nerve fibres positive for the sensory neurone specific sodium channel SNS/PN3, which is regulated by NGF, and may mediate inflammation-induced hypersensitivity. Keratinocyte trk A expression (which mediates an autocrine role for NGF) was increased in clinically affected and unaffected skin, suggesting a compensatory mechanism secondary to reduced NGF secretion at both sites. We conclude that decreased NGF- and SNS/PN3-immunoreactivity, and loss of intra epidermal innervation, may be found without sensory loss on quantitative testing in clinically-unaffected skin in leprosy; this appears to be a sub-clinical change, and may explain the lack of cutaneous pain with inflammation. Sensory loss occurred with reduced sub-epidermal nerve fibres in affected skin, but these still showed trk A-staining, suggesting NGF treatment may restore pain sensation. PMID- 10692625 TI - Intensity dependence of auditory evoked cortical potentials in migraine families. AB - Intensity dependence of auditory evoked cortical potentials is abnormal in migraine. This study investigated intensity dependence in migraine and healthy families using group comparisons and analysis of individual differences. Migraineurs were characterized by a steeper amplitude/stimulus function slope and more pronounced difference between the amplitudes of N1-P2 on the more and the less intensive tones than healthy age matched subjects. Apart from migraine, the age of the participants was an important predictive variable of intensity dependence. Analysis of individual differences revealed low sensitivity and moderate specificity of intensity dependence for migraine. Familial prevalence of intensity dependence among first-degree relatives in migraine families was equal to that in healthy families. These findings support the assumption that high intensity dependence reflects a functional CNS trait which is more pronounced and prevalent in migraine, but may also be found in healthy individuals and in other neuropsychiatric disorders. Increased intensity dependence is only one of several factors contributing to the risk for this form of headache. PMID- 10692626 TI - Delta opioid receptor mediated actions in the rostral ventromedial medulla on tail flick latency and nociceptive modulatory neurons. AB - The rostral ventromedial medulla (RVM) is critical for the modulation of dorsal horn nociceptive transmission. Three classes of RVM neurons (ON, OFF, and NEUTRAL) have been described that have distinct responses to noxious stimuli and mu opioid receptor (MOR) agonists. The present study in barbiturate anesthetized rats investigated the effects of the delta 2 opioid receptor (DOR2) agonist, [D Ala2]deltorphin II (DELT), microinfused into the RVM on the tail flick reflex and activity of RVM neurons. Tail flick latencies increased dose-dependently after administration of DELT (0.6 nmol and 1.2 nmol). Furthermore, DELT inhibited the tail flick related increase in ON cell activity and shortened the tail flick related pause in OFF cell activity. The activity of NEUTRAL cells was not affected. The antinociceptive effects and corresponding changes in ON and OFF cell activity produced by DELT were antagonized by the DOR2 antagonist, naltriben methanesulfonate, administered at the same site. These DOR2 mediated effects on noxious stimulation-evoked changes in RVM neuronal activity are similar to those reported for MOR agonists and suggest that both DOR2 and MOR produce analgesia through activation of OFF cells. PMID- 10692627 TI - Dorsal column-thalamic pathway is involved in thalamic hyperexcitability following peripheral nerve injury: a lesion study in rats with experimental mononeuropathy. AB - A total of 68 neurons were recorded from the ventro-postero-lateral nucleus of thalamus (VPL) in rats with a unilateral chronic constriction injury (CCI) of the sciatic nerve (n=20), sham operation (n=24) and naive rats (n=24), and effects of the lesion of dorsal column (DC) pathway [DC lesion or DC+gracile nucleus lesions] on VPL nucleus neuronal activities were studied. In the VPL nucleus contralateral to the CCI (receiving input from the injured nerve), response latencies of low threshold mechanoreceptive (LTM) and wide dynamic range (WDR) neurons to electrical stimulation of the sciatic nerve were significantly longer than that in the contralateral VPL nucleus receiving input from the sham-operated side (P<0.05). In contrast, response latencies of LTM and WDR neurons to DC stimulation were not different between the sham operated and CCI sides (0.05). Background activity of WDR neurons was significantly higher in the VPL nucleus contralateral to the CCI side when compared to neurons in the VPL nucleus contralateral to the sham operated side and in naive animals. Responses of LTM and WDR neurons to innocuous mechanical stimulation of the receptive fields were significantly decreased after DC and DC+gracile nucleus lesions in all animals. However, the responses of WDR neurons to noxious stimuli were selectively reduced only in rats with CCI by DC and DC+gracile nucleus lesions (P<0.05). The decrease in noxious stimulus-evoked responses of WDR neurons in the VPL nucleus contralateral to the CCI side after DC and DC+gracile nucleus lesions was greater than that in the VPL nucleus contralateral to the sham operated side and naive animals. These results indicated that DC and DC+gracile nucleus lesions produced selective and stronger effect on noxious responses of VPL nucleus WDR neurons receiving input from the site of nerve injury. The findings suggest that the gracile nucleus-thalamic pathway conveys, or modulates, nociceptive information to the VPL nucleus following peripheral nerve injury, resulting in an increase in VPL nucleus response to noxious stimuli that contributes to the development of mechanical hyperalgesia. PMID- 10692628 TI - Prolonged ovarian sex steroid treatment of male rats produces antinociception: identification of sex-based divergent analgesic mechanisms. AB - Simulation of the pregnancy blood concentration profile of 17beta-estradiol (E(2)) and progesterone (P) in nonpregnant ovariectomized rats has been shown to result in a significant elevation of nociceptive response thresholds. The present report demonstrates that spinal opioid antinociceptive responsiveness to these ovarian steroids is not sex-specific. Treatment of orchidectomized sexually mature males with an analogous regimen of E(2) and P also elicits an antinociception, the robustness and temporal profile of which is comparable with that previously observed in females. Neither E(2) nor P, alone, is sufficient to produce antinociception in male rats, as was previously demonstrated in females. Neurobiological substrates and antinociceptive mechanisms underlying ovarian sex steroid antinociception do, however, exhibit sex specificity. In males, the analgesia resulting from ovarian steroid treatment derives from the independent contributions of spinal kappa and mu, not delta, opioid receptor pathways that are additive, not synergistic. Spinal alpha(2)-noradrenergic receptor activity and its attendant analgesic synergy with spinal opioid systems do not contribute to ovarian sex steroid analgesia in males. This is in contrast to the previous demonstrations that ovarian sex steroid-induced antinociception in females results from antinociceptive synergy between activated spinal kappa/delta opioid as well as alpha(2)-noradrenergic receptor systems. The current data reveal that ovarian steroid-activated multiplicative spinal antinociceptive pathways that had been demonstrated in female rats are not manifest in their male counterparts. PMID- 10692629 TI - Debilitating chronic pain syndromes after presumed intraneural injections. AB - This report presents seven patients with severe disability established at the time of a peripheral nerve block. In most of the cases, the injection was administered as a routine procedure by an experienced anesthesiologist. The patient histories suggest that the condition, which can be resistant to all treatment, in most cases could have been avoided if careful attention had been given to the occurrence of pain during the nerve block. It is likely that the risk of devastating iatrogenic disability can be minimized if a few basic principles are respected during the administration of peripheral nerve blocks. PMID- 10692630 TI - Ziconotide for the treatment of severe spasticity after spinal cord injury. AB - Spasticity is a major clinical manifestation of spinal cord injury and upper motor neuron syndrome. PMID- 10692631 TI - Adverse effects associated with the intrathecal administration of ziconotide. AB - The omega-conopeptide, ziconotide, is an N-type calcium-channel blocker that has been shown to produce antinociception in animals using formalin and hot-plate tests. Initial reports of intrathecal administration of ziconotide in cancer and AIDS patients whose pain was unrelieved with opioids demonstrated analgesic efficacy. Although adverse effects were reported, these appeared to be easily managed through dose reduction or symptomatic treatment. This clinical report describes the experiences of three patients with serious adverse effects associated with intrathecal ziconotide. PMID- 10692632 TI - Electroconvulsive therapy for phantom limb pain. AB - Phantom limb pain is common in amputees. Although several treatments are available, a significant number of patients are refractory. Electroconvulsive therapy (ECT), which is usually given to patients with psychiatric disorders such as major depression, has shown efficacy in patients with a variety of pain syndromes occurring along with depression. Two patients are described herein with severe phantom limb pain refractory to multiple therapies, without concurrent psychiatric disorder, who received ECT. Both patients enjoyed substantial pain relief. In one case, phantom pain was still in remission 3.5 years after ECT. It is concluded that phantom limb patients who are refractory to multiple therapies may respond to ECT. PMID- 10692633 TI - Complete Freund's adjuvant-induced hyperalgesia: a human perception. AB - Much of our current understanding about chronic pain and the mechanisms of nociception has been derived from animal models (Bennett GJ. Animal models of neuropathic pain. In: Gebhart, GF, Hammond DL, Jensen TS, editors. Progress in pain research and management, vol. 2, Proceedings of the 7th World Congress of Pain. Seattle, WA: IASP Press, 1994. pp. 495-510; Dubner R, Methods of assessing pain in animals. In: Wall PD, Melzack R, editors. Textbook of pain, vol. 3. Edinburgh: Churchill Livingstone, 1994. pp. 293-302). It has been argued in some cases that animals do not perceive 'pain' as humans do, and thus extrapolation of the results of studies in animals is invalid. Clearly, the animal models used in the laboratory do not approach the complexity of chronic pain encountered in the clinical setting. Human pain perception is more complex since it encompasses lesion variability, as well as psychosocial, cultural, developmental, and environmental variables. Where parallels exist, it is possible to gain insight into certain aspects of human pain syndromes that are likely to lead to improved therapeutic opportunities for individual patients. One such model that is frequently used in animals to study pain associated with inflammation is the subcutaneous injection of complete Freund's adjuvant (CFA). For ethical reasons, however, little information is available from humans concerning pain associated with this form of inflammation. Due to an inadvertent subcutaneous injection of CFA into the terminal phalanx of this investigator, a study with an N of 1, was conducted to compare the subjective effects of CFA-induced inflammation with objective measurements. PMID- 10692634 TI - Virtual reality as an adjunctive pain control during burn wound care in adolescent patients. AB - For daily burn wound care procedures, opioid analgesics alone are often inadequate. Since most burn patients experience severe to excruciating pain during wound care, analgesics that can be used in addition to opioids are needed. This case report provides the first evidence that entering an immersive virtual environment can serve as a powerful adjunctive, nonpharmacologic analgesic. Two patients received virtual reality (VR) to distract them from high levels of pain during wound care. The first was a 16-year-old male with a deep flash burn on his right leg requiring surgery and staple placement. On two occasions, the patient spent some of his wound care in VR, and some playing a video game. On a 100 mm scale, he provided sensory and affective pain ratings, anxiety and subjective estimates of time spent thinking about his pain during the procedure. For the first session of wound care, these scores decreased 80 mm, 80 mm, 58 mm, and 93 mm, respectively, during VR treatment compared with the video game control condition. For the second session involving staple removal, scores also decreased. The second patient was a 17-year-old male with 33.5% total body surface area deep flash burns on his face, neck, back, arms, hands and legs. He had difficulty tolerating wound care pain with traditional opioids alone and showed dramatic drops in pain ratings during VR compared to the video game (e.g. a 47 mm drop in pain intensity during wound care). We contend that VR is a uniquely attention-capturing medium capable of maximizing the amount of attention drawn away from the 'real world', allowing patients to tolerate painful procedures. These preliminary results suggest that immersive VR merits more attention as a potentially viable form of treatment for acute pain. PMID- 10692635 TI - Potassium transport in fungi and plants. PMID- 10692636 TI - Regulation of plasma membrane H(+)-ATPase in fungi and plants. AB - The plasma membrane H+-ATPase from fungi and plants is a proton pump which plays a key role in the physiology of these organisms controlling essential functions such as nutrient uptake and intracellular pH regulation. In fungal and plant cells the activity of the proton pump is regulated by a large number of environmental factors at both transcriptional and post-translational levels. During the last years the powerful tools of molecular biology have been successfully used in fungi and plants allowing the cloning of a wide diversity of H+-ATPase genes and rapid progress on the molecular basis of reaction mechanism and regulation of the proton pump. This review focuses on recent results on regulation of plasma membrane H+-ATPase obtained by molecular approaches. PMID- 10692637 TI - Topology and transport of membrane lipids in bacteria. AB - The last two decades have witnessed a break-through in identifying and understanding the functions of both the proteins and lipids of bacterial membranes. This development was parallelled by increasing insights into the biogenesis, topology, transport and sorting of membrane proteins. However, progress in research on the membrane distribution and transport of lipids in bacteria has been slow in that period. The development of novel biochemical in vitro approaches and recent genetic studies have increased our understanding of these subjects. The aim of this review is to present an overview of the current knowledge of the distribution and transport of lipids in both Gram-positive and Gram-negative bacteria. Special attention is paid to recently obtained results, which are expected to inspire further research to finally unravel these poorly understood phenomena. PMID- 10692638 TI - Recent progress in sustained/controlled oral delivery of captopril: an overview. AB - The development of oral sustained or controlled release dosage form of captopril has been an interested topic of research for a long period of time. Difficulties encountered with such topic based on the fact that the drug is freely water soluble and for obvious reasons, such drug is difficult to be delivered orally in a sustained or controlled release manner and, on another hand, the drug is unstable in the alkaline pH of the intestine, which enabled the scientists to localize the developed formulations at the targeted areas of the GIT. Due to its effectiveness and intensive use as a drug of choice in the treatment of hypertension and congestive heart failure, numerous sustained and controlled release formulations of captopril have been made and reported. Despite of these numerous attempts and works, very few have been successful and some of these formulations have been patented. The clinical supportive data regarding the efficacies of these developed formulations are not always available and, furthermore, their claims rely only on in vitro data. PMID- 10692639 TI - Studies of polymorphism in three compounds by single crystal X-ray diffraction. AB - The role of single crystal diffraction in the quantitative determination of polymorphism is demonstrated by the examination of three compounds. Two polymorphs were found for each of the compounds bis(2-nitrophenyl) trisulphide (1), 2-amino-5-nitrobenzophenone (2) and bis(2-nitrophenyl) sulphide (3). Only in one polymorph of (1) does molecular symmetry correspond with crystallographic symmetry. In (2) the polymorphs arise in the same crystal class and in the same crystallographic space group whereas in (3) the two polymorphs exist in different crystal classes and hence in different space groups. Crystallographic space group transformation is also discussed. PMID- 10692640 TI - TR146 cells grown on filters as a model of human buccal epithelium: IV. Permeability of water, mannitol, testosterone and beta-adrenoceptor antagonists. Comparison to human, monkey and porcine buccal mucosa. AB - The objective of the present study was to evaluate the TR146 cell culture model as an in vitro model of human buccal epithelium. For this purpose, the permeability of water, mannitol and testosterone across the TR146 cell culture model was compared to the permeability across human, monkey and porcine buccal mucosa. Further, the permeability rates of ten beta-adrenoceptor antagonists (acebutolol, alprenolol, atenolol, labetalol, metoprolol, oxprenolol, pindolol, propranolol, timolol and tertatolol) across the TR146 cell culture model and porcine buccal mucosa were related to their lipophilicity (logD(oct; 7.4)) and capacity factor (k') and to their polar water accessible surface area (PWASA). For water, mannitol, testosterone and some of the beta-adrenoceptor antagonists, the permeability enhancement across the TR146 cell culture model in the presence of sodium glycocholate (GC) was determined. The mannitol and testosterone permeability across the TR146 cell culture model could be related to the permeability across porcine and human buccal mucosa. The permeability of the beta adrenoceptor antagonists across the TR146 cell culture model varied between 2.2 x 10(-6) cm/s (atenolol) and 165 x 10(-6) cm/s (metoprolol). For propranolol the cellular permeability value (P(c)) was lower than expected, probably due to accumulation in the TR146 cell layers. Limited correlation of permeability with k' was observed both for the TR146 cell culture model and the porcine buccal mucosa, although the porcine permeability values were approximately 100 times less than the values determined with the TR146 cell culture model. The permeability values were also found to decrease with increasing PWASA. The PWASA value seemed to be more predictable for permeability than k'. The presence of 12.5 mM GC increased the permeability only for the hydrophilic atenolol, which may help explain the mechanism for GC-induced enhancement. The present results indicate that the TR146 cell culture model can be used as an in vitro model for permeability studies and mechanistic studies of human buccal drug delivery of drugs with different lipophilicity. PMID- 10692641 TI - Evaluation of poly(2-hydroxyethyl methacrylate) gels as drug delivery systems at different pH values. AB - Studies of dynamic and equilibrium swelling, structural characterisation and solute transport in swollen poly(2-hydroxyethyl methacrylate) gels (pHEMA) cross linked with tripropyleneglycol diacrylate (TPGDA) were done for a wide range of TPGDA concentrations. The influence of the pH on these pHEMA properties was evaluated. In swelling studies it was found that in changing the pH from 6.5 to 12.0, a large increase in swelling occurred, from approximately 48 to 55%, for the lowest concentration of TPGDA (1 mol%), and from 40 to 80% for the highest concentration (10 mol%). Fourier transform infrared (FTIR) and differential scanning calorimetry (DSC) measurements were made after the equilibrium swelling of the gels at different pH values, to explain these results. The advantage of using these gels as controlled drug delivery systems is illustrated using salicylic acid (SA) as a model drug. The loading and the release of the SA were made at different pH values and the results obtained showed that it is possible to modulate the hydrogel performance by controlling an external factor, the pH at which the drug loading and release were performed. PMID- 10692642 TI - Parameters affecting the efficacy of a sustained release polymeric implant of leuprolide. AB - The objective of this study was to evaluate the formulation parameters critical to the efficacy of an injectable polymeric implant of leuprolide acetate, formed in situ, in suppressing and maintaining serum testosterone levels of animals in the range 0.5 ng/ml for over 90 days. The formulation evaluated contained 45% (w/w) 75/25 poly (DL-lactide-co-glycolide) polymer having an intrinsic viscosity of 0.20 dl/g, dissolved in 55% (w/w) N-methyl-2-pyrrolidone with 3% (w/w) leuprolide acetate added either as a homogeneous solution or a two-part suspension (A/B) system, in which the drug was dispersed within the polymer solution immediately prior to use. The formulation parameters evaluated in this study included polymer molecular weight, polymer concentration, and drug loading. Both rat and dog models were used to evaluate efficacy. Serum testosterone was assayed by radioimmunoassay to determine efficacy, and retrieved implants from the rats at the termination of the study were analyzed by HPLC for residual drug content to determine the extent of drug release. With the candidate formulation, testosterone levels in dogs diminished to the targeted levels of 0. 5 ng/ml by day 14 and remained suppressed up to day 91, reproducing the results seen in rats. Variations in polymer concentration (40-50%), and drug load (3-6% (w/w)) did not have a significant effect on the apparent level and duration of efficacy. However, employing lower molecular weight polymer decreased the duration of efficacy of the formulation. PMID- 10692643 TI - Intestinal first-pass effect of bumetanide in rats. AB - The intestinal first-pass effect of bumetanide was investigated after intravenous and intraportal infusion, and intragastric and intraduodenal instillation of the drug to rats. The AUC(0-->8 h) values of bumetanide after intragastric and intraduodenal instillation of the drug, 10 and 20 mg/kg, were significantly smaller than AUC values after intraportal administration, suggesting that the gastrointestinal first-pass effect of bumetanide was considerable in rats. However, the AUC(0-->8 h) values of bumetanide between intragastric and intraduodenal instillation were comparable, suggesting that the gastric first pass effect of bumetanide was almost negligible in rats. The AUC(0-->8 h) values of bumetanide after intraduodenal instillation were significantly smaller than AUC values after intraportal infusion at 10 (89.8 vs 569 microg min per ml) and 20 (304 vs 1230 microg min per ml) mg/kg, indicating that the first-pass organ(s) of bumetanide was intestine. The F values were 15.8 and 24.7% after intraduodenal instillation of bumetanide, 10 and 20 mg/kg, respectively. Approximately 76.1 and 76.5% of intraduodenally instilled bumetanide disappeared (as a result of absorption and first-pass effect) after 10 and 20 mg/kg, respectively. Therefore, it could be concluded that approximately 60. 3 and 51.8% of the oral dose of bumetanide disappeared by intestinal first-pass effect at 10 and 20 mg/kg, respectively. PMID- 10692644 TI - Lecithin vesicular carriers for transdermal delivery of cyclosporin A. AB - Two kinds of vesicles with and without the presence of sodium cholate (flexible vesicles and conventional vesicles) were prepared, using cyclosporin A as model drug. When applied onto the excised abdominal skin of mice non-occlusively, the enhancing effects of vesicles on the penetration of cyclosporin A were assessed by an in vitro permeation technique. The effect of sodium cholate micelles was also studied. In vivo study was carried out by topical application of vesicles onto the mice skin and drug serum concentration was detected. Results showed that after 8 h of administration, flexible vesicles transported 1.16 microg of cyclosporin A through per cm(2) mice skin and amounted to 1.88 microg 24 h later. The residual amount in the skin was 1.78+/-0.51 microg/cm(2). However, flexible vesicles failed to transport measurable amount of drug through pre-hydrated skin while deposited 2.39+/-0.26 microg/cm(2) into the skin. Conventional vesicles failed to transfer cyclosporin A into the receiver while accumulated 0. 72+/-0.19 microg/cm(2) of drug in the skin. Furthermore, 1 and 40% sodium cholate micelles precluded the transport of cyclosporin A. In vivo studies indicated that with the application of flexible vesicles, serum drug concentration of 53.43+/-9.24 ng/ml was detected 2 h later. After the stratum corneum of mouse skin has been destroyed by shaving, flexible vesicles transferred large amount of drug into blood, up to 187.32+/-53.21 ng/ml after 1 h of application. Conventional vesicles failed to deliver measurable amount of drug into the blood under normal skin condition. In conclusion, flexible vesicle is better than conventional vesicle as the carrier for transdermal delivery of cyclosporin A. Penetration and fusion have been suggested to be two major functional mechanisms. Hydration is detrimental to the enhancement effect. Stratum corneum constitutes main barrier to the transport of lipophilic cyclosporin A. PMID- 10692645 TI - Cellular transport processes of aminoguanidine, a nitric oxide synthase inhibitor, in the opossum kidney cell culture line. AB - Aminoguanidine has potential pharmacologic utility for diabetes and nitric oxide mediated inflammation. Because aminoguanidine is positively charged at physiologic pH (pK(a) approximately 10), it is unlikely that simple diffusion is a predominant mechanism for cellular penetration. This study sought to determine the transport processes by which aminoguanidine, a cationic compound, traverses across cellular membranes. In cultured opossum kidney (OK) cell monolayers, aminoguanidine transport involved both saturable and non-saturable diffusion processes. At passage numbers below 67, the observed V(max) and K(m) for saturable influx were significantly lower than that observed at passages greater than 79 (V(max): low passage, 21.2+/-7.8 pmol/(min*mg protein), n=3; versus high passage, 129.7+/-24.3 pmol/(min*mg protein), n=3, P<0.05; K(m): low passage, 23.7+/-10.8 microM, n=3; versus high passage, 101.7+/-5.6 microM, n=3, P<0.05; mean+/-S.E.M.). Nonsaturable processes were not statistically different (k(ns): low passage, 1.6+/-0.1 pmol/(min*mg protein*microM), n=3; high passage, 1.1+/-0.2 pmol/(min*mg protein*microM) n=3). Saturable influx was temperature dependent, and independent of ATP energy, sodium gradients or changes in membrane potential. Other organic cations competitively inhibited and trans-stimulated saturable influx. Aminoguanidine influx was increased in the presence of an outwardly directed proton gradient and was inhibited in the presence of an inwardly directed proton gradient. Correspondingly, aminoguanidine efflux was trans79) express a saturable, bi-directional carrier-mediated process to transport aminoguanidine across cellular membranes. PMID- 10692646 TI - Pharmacokinetic changes of cyclosporine after intravenous and oral administration to rats with uranyl nitrate-induced acute renal failure. AB - The effects of renal failure on the pharmacokinetics of cyclosporine were investigated after intravenous, 30 mg/kg, and oral, 100 mg/kg, administration of the drug using a rat model of uranyl nitrate-induced acute renal failure (U-ARF). After intravenous administration to rats with U-ARF, the volume of distribution at steady state (1.97 vs. 2.56 l/kg) was significantly smaller, and the area under the blood concentration-time curve (348 vs. 296 microg h/ml) tended to be greater and total body clearance (0.0851 vs. 0. 102 l/h per kg) tended to be slower than those in control rats. After oral administration, the pharmacokinetic parameters were not significantly different between the control rats and rats with U-ARF, suggesting that U-ARF did not considerably affect the pharmacokinetics of cyclosporine after oral administration. PMID- 10692647 TI - Influence of the acid type on the physical and drug liberation properties of chitosan-gelatin sponges. AB - The influence of acid type used to dissolve chitosan on the resulting sponge physical properties, and their consequent effect on the drug liberation were investigated. Chitosan was dissolved in different acid solutions and chitosan gelatin sponges were produced by frothing up the polymer solution and then freeze drying the foam. Prednisolone was used as a model drug. Using tartaric or citric acid resulted in instable, soft, elastic and disintegrating sponges with fast drug release. Elastic but harder sponges from stable foams were obtained when hydrochloric or lactic acid were used. The use of acetic or formic acid enabled the production of stable foams, soft and elastic sponges and a slow drug release. The rate of drug release was decreased by crosslinking the polymers with glutaraldehyde, but only if acetic, formic or acetic acid were used. Therefore, it is possible to manipulate the mechanical properties and the drug liberation rate by using different acids to dissolve chitosan. PMID- 10692648 TI - Synthesis and characterisation of mucoadhesive thiolated polymers. AB - This study examined various factors influencing the mucoadhesive properties of thiolated polymers (thiomers), which are capable of forming covalent bonds with thiol sub-structures of the mucus glycoprotein. Mediated by a carbodiimide, L cysteine was therefore covalently bound to polycarbophil (PCP) and to carboxymethylcellulose (CMC). The resulting polymer conjugates displayed 12.3 and 22.3 micromol thiol groups per gram, respectively. Whereas the swelling behaviour of tablets based on CMC was not markedly influenced by the immobilisation of cysteine, it was improved significantly (P<0.05) in case of PCP. Tensile studies carried out with the unmodified and thiolated polymers of pH 3, 5 and 7, respectively, revealed that only if the polymer displays a pH-value of 5, the total work of adhesion can be improved significantly due to the covalent attachment of thiol groups. These results were in good agreement with a new mucoadhesion test system described here taking also the cohesiveness of the delivery system into account. The results represent helpful basic information in order to improve the mucoadhesive properties of thiolated polymers. PMID- 10692649 TI - A comparative in vitro study of percutaneous penetration of beta-blockers in human skin. AB - In vitro diffusion experiments with propranolol, oxprenolol, metoprolol and atenolol were carried out using excised human abdominal skin. The main permeation parameters (permeability coefficient, flow and lag time) were calculated and compared as measurement of intrinsic permeability across human skin. A long lag time and a low steady-state flow were found for all drugs assayed. Skin permeability predicted at steady state did not reach therapeutic concentrations, which indicated the need for appropriate chemical penetration enhancers or vehicles to overcome limiting factors. The results, including those of celiprolol and bisoprolol reported previously, correlated with physicochemical properties, especially with lipophilicity, one of the main factors in drug permeability prediction through human skin. PMID- 10692650 TI - In vitro biocompatibility studies with the experimental anticancer agent BIBX1382BS. AB - The novel anticancer agent BIBX1382BS is a representative of the human epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors. BIBX1382BS, for parenteral use, is formulated pharmaceutically as a lyophilized product containing 100 mg BIBX1382BS per dosage unit. This in vitro study was performed to establish the optimal intravenous administration conditions (infusion concentration and infusion rate) for the forthcoming clinical absolute oral bioavailability study of BIBX1382BS. BIBX1382BS infusion solutions have a low pH in order to keep the substance in solution. We therefore decided to investigate the hemolytic and precipitation potential of the drug in vitro. Also, the ratio of formulation (F) solution volume and a blood simulans (B) volume necessary to reach the physiological pH, expressed as the FB-ratio, was determined in vitro. On the basis of the results obtained, it is advised to administer BIBX1382BS infusion at a concentration of 1 mg/ml and a maximum infusion rate of 10 ml/min. This article describes the in vitro biocompatibility screening program. PMID- 10692651 TI - The role of the cardiovascular specialist in the prevention of cardiovascular diseases - executive summary. PMID- 10692652 TI - The global burden of cardiovascular diseases. PMID- 10692653 TI - The burden of cardiovascular diseases in Canada. PMID- 10692654 TI - Prevention and control of risk factors among children and youth. PMID- 10692655 TI - Women and cardiovascular diseases. PMID- 10692656 TI - Cardiovascular disease prevention in the elderly. PMID- 10692657 TI - Cardiovascular diseases and aboriginal peoples. PMID- 10692658 TI - Cardiovascular diseases in ethnic groups. PMID- 10692659 TI - Lipids and cardiovascular diseases. PMID- 10692660 TI - The 1999 Canadian recommendations for the management of hypertension. On behalf of the Task Force for the Development of the 1999 Canadian Recommendations for the Management of Hypertension. PMID- 10692661 TI - Obesity and cardiovascular diseases. PMID- 10692662 TI - Emerging risk factors associated with cardiovascular diseases. PMID- 10692663 TI - Smoking. PMID- 10692664 TI - Diet and the prevention of cardiovascular diseases. PMID- 10692665 TI - Physical inactivity. PMID- 10692666 TI - Psychosocial risks and cardiovascular diseases. PMID- 10692667 TI - Socioeconomic factors and social support. PMID- 10692668 TI - Risk factors for stroke. PMID- 10692669 TI - Peripheral arterial disease. PMID- 10692670 TI - Cardiac rehabilitation programs. PMID- 10692671 TI - The cost effectiveness of preventing cardiovascular diseases. PMID- 10692672 TI - Partnerships in cardiovascular disease prevention. PMID- 10692673 TI - Apoptosis, bcl-2 expression and p53 accumulation in myelodysplastic syndrome, myelodysplastic-syndrome-derived acute myelogenous leukemia and de novo acute myelogenous leukemia. AB - Apoptosis and its dysregulation have been implicated in dysplastic and ineffective hematopoiesis and the neoplastic transformation of bone marrow in myelodysplastic syndrome (MDS). To explore the role of apoptosis in hematological disorders, we examined the frequency of apoptotic cells by the in situ end labeling method in bone marrow specimens from 37 patients with MDS [refractory anemia (RA) 10 cases, RA with excess of blasts (RAEB) 27 cases including 12 cases with leukemic transformation], 12 patients with MDS-derived acute myelogenous leukemia (AML) and 13 patients with de novo AML. In addition, we investigated the relationship of apoptosis to the immunohistochemical expression of bcl-2 and p53 in these cases, and the association of apoptosis, bcl-2, and p53 with the leukemic evolution of MDS by examining sequential bone marrow samples of the same patient from the time of initial diagnosis to the time of overt leukemia. The percentage frequency of apoptotic cells was significantly greater in MDS (RA: 9.46 +/- 2.99%, m +/- SD; RAEB: 5. 60 +/- 3.09) as compared with those in MDS derived AML (0.62 +/- 0. 37), de novo AML (0.28 +/- 0.11) and controls (1.00 +/- 0.59). On the other hand, the cases of RAEB with leukemic transformation exhibited a lower frequency of apoptotic cells and a higher frequency of bcl-2- and p53-positive cells than those without transformation. When the RAEB cases transformed to AML, the frequency of apoptotic cells was significantly reduced (2.96 +/- 1. 54 --> 0.62 +/- 0.37), while the frequencies of bcl-2-positive cells and p53-positive cells were greater (10.88 +/- 3.66 --> 30.54 +/- 7. 14, and 20.21 +/- 6.21 --> 32.34 +/- 14.71, respectively). In contrast to MDS-derived AML, over a half of de novo AML cases showed few p53-positive cells. These findings corroborate the earlier notion that apoptosis may play a substantial role in dysplastic and ineffective hematopoiesis in MDS. It is also suggested that the suppression of apoptosis associated with enhanced bcl-2 expression and p53 accumulation increases the probability of developing leukemia in MDS, and that oncogenetic development might be different between MDS-derived AML and de novo AML. PMID- 10692674 TI - Function and X chromosome inactivation analysis of B lymphocytes in myelodysplastic syndromes with immunological abnormalities. AB - To investigate the pathogenesis of immunological abnormalities (IA) in myelodysplastic syndrome (MDS), we examined B cells for their ability to produce cytokine and their X chromosome inactivation pattern (XCIP). An IA was defined as being positive for at least one autoimmune laboratory test (e.g. antinuclear antibody, rheumatoid factor). Seventy-three MDS patients [65 with refractory anemia (RA), 3 with RA with excess blasts (RAEB), and 5 with RAEB in transformation] were examined; 47 had IA and 26 had no IA. To examine the function of B cells in MDS, the production of interleukin (IL)-6 and IL-10 was measured in cultures of purified B cells with or without stimulators. Both IL-6 and IL-10 production rates in patients with IA were significantly higher than in patients without IA and normal controls. The skewing of XCIP of B cells was analyzed by using the polymerase chain reaction, and the skewing rate of B cell XCIP was quantitatively assayed by compared to control T lymphocytes. The skewing rate of B cells was higher in patients with IA than in those without IA and normal controls. Therefore, a small population of B cells in patients with IA might be derived from MDS clones, and be associated with the induction of IA. PMID- 10692675 TI - Plasma thrombopoietin concentrations in response to platelet transfusion therapy. AB - Studies performed in rabbit and mouse models and in a limited number of human subjects, show that transfused platelets bind thrombopoietin (TPO) and decrease its concentration in the circulation. The aim of the present study was to further examine this relationship. The material comprised 12 patients receiving a total of 21 transfusions, as part of the routine clinical treatment. Blood samples were collected from the patients immediately before and 30 min after completion of the platelet transfusion, and the corrected platelet count increment (CCI) was calculated. A commercially available ELISA kit was used to determine plasma TPO concentrations. Statistically significant reductions in median TPO concentration were observed in response to the platelet transfusions. Patients who were refractory to platelet transfusions showed the slightest decrease in TPO concentration. As for the linear regression between change in TPO level and CCI, only borderline significance was observed. Thus, our findings support the concept that platelets can remove TPO from the circulation. PMID- 10692676 TI - The synergistic effect of thrombopoietin in erythropoiesis and myelopoiesis from human bone marrow cells. AB - We sought to determine whether recombinant human thrombopoietin (TPO) acts synergistically with recombinant human erythropoietin (EPO) and/or recombinant human interleukin-3 (IL-3) on erythroid burst formation and granulocyte macrophage colony formation from human bone marrow (BM). BM cells were from 5 adults and 15 children who underwent bone marrow examination because of a clinical suspicion of malignancy; their bone marrows as well as the complete blood counts were normal and were cultured in a methylcellulose system. TPO has a synergistic effect with EPO or EPO + IL-3 on erythropoiesis of human BM, as the addition of TPO to EPO significantly gave rise to more erythroid bursts (p = 0.0001) and the addition of TPO to EPO + IL-3 might give rise to more erythroid bursts (p = 0.05). TPO also has a synergistic effect with recombinant human granulocyte colony-stimulating factor (G-CSF) on myelopoiesis of human BM, since the addition of TPO to G-CSF gave rise to significantly more granulocyte macrophage colonies (p = 0. 0001). Besides its well-known significant role in megakaryopoiesis, TPO also has synergistic effects on erythropoiesis and myelopoiesis. PMID- 10692677 TI - GM-CSF in sickle cell anemia patients with elevated Hb F. AB - We estimated plasma GM-CSF levels in a group of 28 steady-state sickle cell anemia (SS) patients in Kuwait, using an ELISA technique. There were 24 age matched Hb AA controls, 14 of whom were healthy while 10 were acutely ill at the time of the study. Five SS patients were also studied during 6 episodes of painful crisis. Among the SS patients, 82.1% were homozygous for the Saudi Arabia/India (SAI) haplotype with Hb F ranging from 15 to 35% and total Hb from 8.5 to 11 g/dl. Three patients (siblings) were SAI/Benin compound heterozygotes with Hb F of 9-23% and total Hb >10 g/dl. One patient each was homozygous for the Benin or the Bantu haplotype; they had Hb F <2% and total Hb of 6.6 and 7.2 g/dl, respectively. Four (14. 3%) steady-state SS patients had detectable plasma GM-CSF ranging from 75 to 1,817.6 pg/ml. These included the 2 patients with Hb F <2. 0% and 2 with the SAI/Benin compound heterozygotes with Hb F of 11 and 9%, respectively. Four (66.7%) SS patients in crisis, 6 (42.9%) healthy controls and 6 (60%) acutely ill controls had detectable plasma GM-CSF. A clearcut association of GM-CSF with Hb F level or degree of anemia in steady-state SS patients could not be established. The appearance of GM-CSF in the plasma of patients in crisis and also among control subjects raises the possibility that other factors are involved in the production of this cytokine in the subjects studied. PMID- 10692678 TI - Efficacy of further attempts to mobilize CD34+ peripheral stem cells with alternative procedures after primary failure. AB - 19 patients who failed the target collection of at least 2.5 x 10(6) CD34+ cells/kg underwent further mobilization procedures either with granulocyte-colony stimulating factor (G-CSF) alone after failure to chemotherapy plus G-CSF (group 1), or with chemotherapy plus G-CSF (group 2), or with high-dose G-CSF (24 microg/kg) alone (group 3) after failure to respond to standard dose of G-CSF (10 microg/kg) alone. In all groups, an increase in median CD34+ cell yield could be observed following alternative procedures (1.1- to 1.9 x 10(6) kg; p = 0.02). The highest increase in CD34+ cell harvest was achieved in group 1 (0.85 to 2.2 x 10(6) kg), followed by group 2 (1. 2 to 1.7) and group 3 (1.0 to 1.4), but without statistically significant difference between the mobilization technologies. All patients with more than 1.0 x 10(6) CD34+ cells/kg in the first apheresis procedure reached the overall target of 2.5 x 10(6) CD34+ cells/kg after a second or subsequent mobilization procedure. In contrast, only 3 of 8 patients (37%) with less than 1.0 x 10(6) CD34+ cells in the first harvest could reach the target of 2.5 x 10(6) CD34+ cells after further mobilization attempts. PMID- 10692679 TI - Effect of ozone on red blood cell enzymes and intermediates. AB - Ozone autohemotherapy has been considered a form of alternative medicine and has not yet been subjected to the rigors of well-designed clinical trials. Despite encouraging anecdotal reports regarding the use of ozone in various disorders, there has been a concern that ozone per se may adversely affect red cell membranes and metabolites. The purpose of this study was to ascertain the effect of ozone administration at a concentration commonly used in autohemotherapy on a panel of red cell enzymes and intermediates, as well as its effect on red cell integrity. Since these parameters were unaffected by ozone, we suggest that clinical trials for the use of ozone autohemotherapy should be encouraged. PMID- 10692680 TI - Are haemochromatosis mutations related to the severity of liver disease in hepatitis C virus infection? AB - It has been proposed that iron overload may adversely affect liver disease outcome. The recent identification of 2 mutations in the HFE gene related to hereditary haemochromatosis (Cys282Tyr and His63Asp) provided an opportunity to test whether they are associated with hepatic iron accumulation and the activity and severity of liver disease in hepatitis C virus (HCV) infection. We investigated the prevalence of HFE mutations in 135 male patients with chronic HCV hepatitis, and correlated genotype distribution with different parameters of iron status and the activity and severity of liver disease. Of these 135 patients, 6 (4.4%) carried Cys282Tyr and 32 (23.7%) carried His63Asp, frequencies which were similar to those observed in healthy controls. Serum iron levels and transferrin saturation (but not ferritin levels or liver iron content) were significantly higher in carriers than in non-carriers of HFE mutations. No difference was observed in serum ALT, AST and GGT levels between carriers and non carriers. Finally, scores for necroinflammatory activity and fibrosis in the liver were significantly higher in HFE carriers than in non-carriers. Patients with chronic HCV infection carrying HFE mutations tend to present more evident body iron accumulation and a higher degree of necroinflammatory activity and fibrosis in the liver. HFE gene mutations might be an additional factor to be considered among those implicated in the determination of a worse prognosis of the liver disease in chronic HCV infection. PMID- 10692681 TI - Graft failure of autologous peripheral blood stem cell transplantation due to acute metabolic acidosis associated with total parenteral nutrition in a patient with relapsed breast cancer. AB - A 32-year-old female had been diagnosed as having relapsed breast cancer and liver metastasis. She underwent high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (PBSCT) with 5.8 x 10(6)/kg CD34+ cells. She was supported by total parenteral nutrition (TPN) without vitamins throughout these therapies. Hematopoietic recovery was not observed by day 28 after PBSCT, necessitating a second PBSCT on day 29 using the back-up material of 4.4 x 10(6)/kg CD34+ cells. On the next day, she suddenly developed severe metabolic acidosis, heart failure and deep coma. After immediate infusion of thiamine, heart failure and coma rapidly improved. The neutrophil count reached 0.5 x 10(9)/l on day 9 and the platelet count 50 x 10(9)/l on day 15 after the second PBSCT. This is a rare graft failure due to acute metabolic acidosis or thiamine deficiency associated with TPN. PMID- 10692682 TI - Thrombotic thrombocytopenic purpura during interferon alpha treatment for chronic myelogenous leukemia. AB - Thrombotic thrombocytopenic purpura (TTP) and hemolytic-uremic syndrome have recently been observed in patients undergoing interferon alpha (IFN-alpha) therapy. However, the relationship between disease and therapy has not been established, essentially because of concomitant treatment or previous bone marrow transplantation. We present a case of TTP developing during IFN-alpha therapy for chronic myelogenous leukemia. In this case, IFN-alpha seems to be the only etiological agent. PMID- 10692704 TI - The relationship of patient education and hypertension treatment compliance. PMID- 10692706 TI - Status of nurse practitioner practice. Report 1. PMID- 10692705 TI - Athena found or lost: the precepting experiences of mentored and non-mentored nurse practitioner students. PMID- 10692707 TI - Prescriptive authority: a dilemma facing advanced practice nurses. PMID- 10692708 TI - Nutrition information on the Internet. PMID- 10692709 TI - Respiratory syncytial virus: understanding the threat to communities. PMID- 10692710 TI - Management of tension-type headache. AB - Tension-type headache is now the term used to describe headaches that have previously been grouped under various ill-defined headings such as 'tension headache', 'stress headache' and 'muscle contraction headache'. Tension-type headaches are characterized by pain that is mild or moderate in severity, bilateral in distribution, pressing or tightening in quality, and unaccompanied by major systemic disturbance or neurological signs. While many people have mild, infrequent tension-type headaches which they do not regard as an illness, a minority have chronic and often daily symptoms. Here we review the management of tension-type headaches in adults. PMID- 10692711 TI - Management of ectopic pregnancy. AB - Each year, around 11,000 women in the UK (around 1% of all pregnancies) present with an ectopic pregnancy, in which a fertilised ovum implants elsewhere than in the uterine cavity, usually a Fallopian tube. Left untreated, tubal ectopic pregnancy can lead to rupture of the Fallopian tube and life-threatening haemorrhage, and never results in a live birth. Improvements in management have led to a fall in the mortality rate from 2.9 per 1000 ectopic pregnancies in the early 1970s to 0.4 per 1000 in 1994-1996. Here we review the current management of tubal ectopic pregnancy. PMID- 10692712 TI - Tazarotene--a topical retinoid for psoriasis. AB - Retinoids have been given systemically for severe psoriasis for several years. Tazarotene (Zorac-Allergan), the first retinoid to be marketed as a topical preparation for psoriasis, was recently licensed for mild-to-moderate plaque psoriasis involving up to 10% of the body surface area. Here we assess its place in the management of psoriasis. PMID- 10692713 TI - Withdrawing patients from antidepressants. AB - Around 1 in 5 people experience an episode of major depressive illness at some time in their lives, and many experience relapse or recurrence. To prevent early relapse, most experts now recommend several months' treatment for patients with acute episodes of depression. Patients considered at high risk for recurrence of severe depression may need long-term maintenance treatment. Doctors have to advise their patients whether and when it is prudent to stop treatment, and must be alert to potential problems when antidepressants are withdrawn. The two main problems are relapse or recurrence of depression, and effects arising from withdrawal of the drugs themselves. We discuss the withdrawal of antidepressant drug treatment in adults with unipolar depression. PMID- 10692714 TI - Nicotine replacement to aid smoking cessation. AB - Cigarette smoking remains the commonest cause of preventable mortality in the UK, accounting for about 120,000 deaths each year among people aged 35 years or more. In all, smoking-related disease costs a typical health authority around 15 million Pounds a year. It is notoriously difficult to stop smoking but success rates are increased if cigarettes are replaced by nicotine given as a medicine. When reviewing nicotine replacement in 1993, we recommended a "combined approach, using nicotine patches plus advice and support". Since then, other forms of nicotine replacement have become available. Here we discuss current evidence on the efficacy and safety of different forms of nicotine replacement and consider the place of such therapy. PMID- 10692715 TI - Who decides on treatment: the patient versus the state. AB - In March this year, we held our seventh annual symposium entitled: 'Who decides on treatment: the patient versus the State'. Within the NHS, there are many levels at which treatment decisions are made. At one extreme, decisions involve individuals at the point of 'delivery' and so are taken during the consultation and involving the patient and prescriber. At the other extreme, decisions are a national matter and taken by government. The symposium explored the changing environments of decision-making and the contribution of those involved. Here we summarise the talks and incorporate ideas that arose in discussion. PMID- 10692716 TI - Defining optimal dosing for sumatriptan tablets in the acute treatment of migraine. AB - Oral sumatriptan, which is a well tolerated, effective acute treatment for migraine, and is selectively available in different countries in 100 mg, 50 mg, and 25 mg tablets. The first large dose-ranging study compared the 100 mg dose to higher doses (200 mg and 300 mg) and found it to be just as efficacious and better tolerated. The first studies comparing the 100 mg dose to lower doses (25 mg and 50 mg) found them all to be similar in effectiveness and tolerability. However, a larger definitive study found that the 100 mg and 50 mg doses offered better efficacy than the 25 mg dose, whereas the 25 mg and 50 mg doses were better tolerated than the 100 mg dose. Thus the 50 mg dose appears to offer the best ratio of efficacy to tolerability. Many patients, though, prefer or require the 100 mg dose and tolerate it well. Allowed to select dosing themselves, patients tend to migrate to the 100 mg dose. PMID- 10692718 TI - Patient preference for oral sumatriptan 25 mg, 50 mg, or 100 mg in the acute treatment of migraine: a double-blind, randomized, crossover study. Sumatriptan Tablets S2CM11 Study Group. AB - BACKGROUND: Dosing recommendations for oral sumatriptan have ranged from 25 mg to 100 mg. Patient dose preferences are clinically relevant (perhaps moreso than traditional efficacy endpoints) and deserve study. METHODS: A multinational randomized double-blind crossover study was conducted over 18 weeks to assess patient dose preference, efficacy, and tolerability for oral sumatriptan (25 mg, 50 mg, and 100 mg) in the acute treatment of migraine; 257 patients treated three attacks, using a different dose for each. RESULTS: The 100 mg dose was preferred by 35% of patients, 31% the 50 mg dose, and 25% the 25 mg dose. Efficacy and speed of action were the two main reasons given for preferring the higher doses. Compared with the 25 mg dose, the 100 mg and 50 mg doses were significantly more likely to provide headache relief at 2, 3, and 4 h after dosing and complete headache resolution at 3 and 4 h after dosing (P < 0.027). Recurrence rates were similar for the three doses, ranging from 33% to 38%, though the median time to recurrence increased with dose, from 8.5 to 11.8 h. The 25 mg, 50 mg, and 100 mg doses were all well tolerated, with adverse event incidences of 19%, 21%, and 30%, respectively. CONCLUSIONS: Patients preferred the 50 mg and 100 mg doses of oral sumatriptan to the 25 mg dose, and the higher doses were more effective against migraine; however, the 25 mg and 50 mg doses were better tolerated than the 100 mg dose. Though the 50 mg dose probably has the best effectiveness-to tolerability ratio, some patients clearly prefer a higher dose. PMID- 10692717 TI - A double-blind placebo-controlled study assessing the efficacy and tolerability of 50 mg sumatriptan tablets in the acute treatment of migraine. Sumatriptan Tablets S2CM07 Study Group. AB - BACKGROUND: Oral sumatriptan 50 mg has been found to have good efficacy and tolerability in the acute treatment of migraine but has been less well studied than the 100 mg dose. METHODS: This was a double-blind, parallel-group study (Glaxo Wellcome protocol number S2CM07) comparing the efficacy and safety of sumatriptan 50 mg tablets with placebo in the acute treatment of migraine. Patients treated three migraine attacks with study medication; a second, optional dose was available for treating recurrent headache. Of the 560 patients randomized, 485 treated at least one attack, 411 at least two attacks, and 362 three attacks. The primary efficacy measure was the proportion of patients who had obtained complete or almost complete headache relief at 4 h after dosing. RESULTS: For all attacks, a significantly greater proportion of patients experienced headache relief at 4 h with sumatriptan 50 mg tablets than with placebo (59% to 62% versus 32% to 42%; P = 0.005). The same was true at 3 h across all attacks, and at 2 h for attacks 1 and 2 (49% versus 23% and 45% versus 29%, respectively). Although sumatriptan and placebo were associated with similar incidences of recurrence, sumatriptan was associated with a longer time to recurrence. The incidence of adverse events with sumatriptan was similar to that with placebo, and there was no increase in adverse events associated with use of a second dose to treat recurrence. CONCLUSIONS: Sumatriptan 50 mg tablets are well tolerated and efficacious in relieving migraine headache. PMID- 10692719 TI - Patient-selected dosing in a six-month open-label study evaluating oral sumatriptan in the acute treatment of migraine. Sumatriptan Tablets S2CM10 Study Group. AB - BACKGROUND: Dosing recommendations for oral sumatriptan as acute treatment for migraine have ranged from 25 mg to 100 mg. Patient dose preferences have not been studied in a setting mimicking clinical practice. METHODS: In an open-label study evaluating patient acceptance and the relative efficacy and safety of 25 mg, 50 mg, and 100 mg doses of oral sumatriptan over a period of six months, 338 patients treated three migraine attacks with 50 mg sumatriptan and then were allowed to double or halve the dose. After treating another three attacks, they were again allowed to adjust the dose by one level. RESULTS: After migraine attack 3, 37% of patients chose to continue with the 50 mg dose, 50% increased the dose to 100 mg, and 12% decreased it to 25 mg. After attack 6, 8%, 33%, and 58% of patients chose the 25 mg, 50 mg, and 100 mg doses, respectively; only 3% of those taking the 100 mg dose chose to reduce it. Overall, the mean percentages of attacks per patient in which headache relief had been obtained 4 h after dosing were 71%, 71%, and 80% for the 25 mg, 50 mg, and 100 mg doses, respectively. Patients who decreased the dose to 25 mg after attack 3 experienced decreases both in adverse events and percentage of attacks with headache relief, whereas in those who increased the dose to 100 mg, likelihood of headache relief increased but the incidence of adverse events did not. CONCLUSIONS: More patients chose the 50 mg or 100 mg dose than the 25 mg dose. All three doses had similar efficacy and tolerability. PMID- 10692720 TI - Evidence in myocardial infarction. PMID- 10692721 TI - Chronic heart failure: evidence for ACE inhibitors. PMID- 10692722 TI - ACE inhibitors: the evidence in hypertension. PMID- 10692723 TI - Structural basis of LV dysfunction: collagen matrix remodelling and the renin angiotensin-aldosterone system. PMID- 10692724 TI - The role of transcription factors in the pathogenesis of type 2 diabetes. PMID- 10692725 TI - State of current therapy and patient care in Europe. PMID- 10692726 TI - Optimisation of insulin treatment to improve cardiovascular risk. PMID- 10692727 TI - The importance of postprandial hyperglycaemia. PMID- 10692728 TI - The role of insulin treatment in type 2 diabetes. PMID- 10692729 TI - Research advances in the treatment of type 2 diabetes mellitus. PMID- 10692730 TI - American journal owners 2, principled editors 0. PMID- 10692731 TI - Microalbuminuria: should lowering albumin excretion be a therapeutic goal? PMID- 10692732 TI - Leg cramps in the elderly: prevalence, drug and disease associations. AB - To determine the prevalence of leg cramps in elderly outpatients and their association any underlying diseases and concomitant drug intake, we conducted a cross-sectional study using an in-depth questionnaire. A total of 365 patients aged 65 years and over (mean 78.5 years) attending our outpatient clinic participated in the study. The prevalence of leg cramps was 50%. Cramps were commoner in females (56%) than in males (40%). Although reported to occur anytime throughout the 24 hours, cramps were most prevalent at night (62%). In many patients, leg cramps were a long-standing complaint: 20% had been suffering with them for more than 10 years, whereas only 9% of patients reported them first starting within the last six months. Only 73 (40%) sufferers had informed their practitioner; of these, 39 (53%) received treatment, of whom 26 gained benefit. Leg cramps were strongly associated with peripheral vascular disease (odds ratio 2.9, 95% CI 1.89-4.55, p < 0.00001), arthritis (odds ratio 2.26, 95% CI 1.48 3.45, p = 0.0001) and female gender (odds ratio 1.96, 95% CI 1.28-3.03, p = 0.002). Heart failure, hypertension, diabetes mellitus and stroke were not significantly associated. Except for a causal association with analgesic use, no positive association could be shown with any other class of drugs, including diuretics. PMID- 10692734 TI - Does the eradication of Helicobacter pylori cure duodenal ulcer disease in communities with a high prevalence rate? Comparison with long-term acid suppression. AB - The long-term effect of Helicobacter pylori eradication on the natural history of duodenal ulcer has been investigated and compared with long-term acid suppression treatment in an endemic community for infection. Seventy-three patients with endoscopically verified H. pylori positive active duodenal ulcer disease were included in this prospective study. Patients were divided into two groups. Group A patients (n = 39) were given an omeprazole-based triple eradication regimen, while group B patients (n = 34) were given omeprazole alone followed by long-term famotidine 20 mg daily as maintenance treatment. A control endoscopy was performed at the third month of treatment. The bacterium was eradicated in 32 (82%) of group A patients. All patients were followed up for two years and an endoscopy performed at the end of each year. H. pylori recurred in 13 patients and the reinfection rate was 44.8% over two years. Duodenal ulcer recurred in seven of these patients at two years (24.1%). There was a close association between H. pylori reinfection and ulcer relapse. Group B patients remained H. pylori positive during the study and the ulcer recurred in five of these patients (6.6%) despite continuous famotidine treatment. There was no statistically significant difference in ulcer relapse rate between the groups. These results suggested that H. pylori eradication is not an absolute solution for duodenal ulcer disease in high endemic regions and continuous maintenance treatment with H2-receptor antagonists is still an alternative approach in some chronic recurrent cases. PMID- 10692733 TI - A prospective audit of hospital-acquired deep vein thrombosis and pulmonary embolism. AB - The incidence of symptomatic deep vein thrombosis and pulmonary embolism acquired in hospital was studied, and the effectiveness of current thromboprophylaxis was assessed in an open study of 8648 admissions to the Doncaster Royal Infirmary between April and July 1994. On admission, all patients were assessed for their likely risk of thromboembolic problems according to THRIFT criteria. Treatment, prophylaxis, complications and outcome were recorded on discharge. A high risk sub-group was followed up for up to 42 days after discharge. The overall rate of clinically apparent hospital-acquired thromboembolic complications was 0.4% (n = 35). The rate of clinically apparent thromboembolic disease in the high risk group was 2.1% (n = 17). The incidence of thromboembolic problems appeared not to be reduce by prophylaxis apparently even when stratified by risk group. These findings suggest that thromboembolic complications may be less common than would be expected from published literature. Thromboprophylaxis as currently practised within our institution does not seem to affect the incidence of deep vein thrombosis or pulmonary embolism, and these results would appear to argue against a 'blanket' policy for pharmacological thromboprophylaxis. PMID- 10692735 TI - An assessment of the efficacy of atorvastatin in treating patients with dyslipidaemia to target LDL-cholesterol goals: the atorvastatin matrix study. AB - A total of 531 patients from 57 hospital centres across the UK, who had previously been treated with lipid-lowering agents in combination or alone, in whom the degree of cholesterol reduction was insufficient to achieve European Atherosclerosis Society target levels, were treated with atorvastatin over a 12 week period. The dose of atorvastatin (10, 20 or 80 mg/day) was determined by assignment of risk based on entry level cholesterol levels and the presence of other established CHD risk factors. Atorvastatin was successful in achieving target LDL-cholesterol levels in 86% of mild risk patients, 88% of moderate risk patients and 52% of high risk patients. Compliance with atorvastatin was 96% and treatment was well tolerated. This study demonstrates that atorvastatin is effective in achieving target lipid levels in a large proportion of patients and that the dose required can be predicted by an assessment of the patient's risk profile. PMID- 10692736 TI - Tuberculous perforations of the small intestine. AB - The hospital records of 58 patients operated on for tuberculous perforations of the small intestines at our hospital between 1987 and 1996 were reviewed. Clinical features were non-specific in the majority of the patients. Pneumoperitoneum on abdominal radiographs was present in only 28 (48.3%) patients. Forty-five (77.6%) were operated on within 36 hours of perforation. Surgical management consisted of resection and end-to-end anastomosis (n = 45); oval excision of the perforation and transverse anastomosis reinforced with an omental patch (n = 7); ileo-transverse colostomy (n = 5); and peritoneal drainage only (n = 1). There were 17 deaths (29.3%). Adverse prognostic factors were operation beyond 36 hours (p < 0.01), multiple perforations (p < 0.001), and faecal fistula formation (p < 0.01). Mortality was least with early resection and end-to-end anastomosis of the perforated bowel segment. We conclude that a high index of suspicion is essential for early diagnosis and optimal treatment of patients with tuberculous intestinal perforations. PMID- 10692737 TI - Treatment with carvedilol is associated with a significant reduction in microalbuminuria: a multicentre randomised study. AB - Patients with mild to moderate essential hypertension (n = 1570) were enrolled in a large, multicentre, randomised, open-label study designed to evaluate the safety and efficacy of different regimens of carvedilol. Reported here are the effects of carvedilol on microalbuminuria (MAU) in a subset of 876 patients who underwent MAU assessment (i.e. the Micral-Test) at baseline and at week 12. MAU was present at baseline in 245 (28%) of these patients. Despite different magnitudes of blood pressure reduction, improvements in MAU were similar in all groups (range 54-60%), with complete disappearance occurring in 48-55% of patients. The decrease in MAU did not correlate with the magnitude of blood pressure reduction, suggesting a possible renal protective effect exerted by carvedilol independent of blood pressure reduction mediated by beta-blockade and vasodilatation. PMID- 10692738 TI - Attention deficit hyperactivity disorder: possible causes and treatment. AB - The possible causes of attention deficit hyperactivity disorder are considered, and the drugs that may be of benefit in treating it. Accurate diagnosis is essential for effective management. This includes the syndrome itself and associated conditions. Theories of causation are discussed. These involve the anatomical substrate but, equally importantly, the role of neurotransmitters; and the use of tests such as positron emission tomography will undoubtedly increase understanding. Genetic factors also play a part. Options for treatment are considered. Evidence supports the effectiveness of stimulant drugs, such as methylphenidate. Among the justifications for such treatment are the excessive strains placed on family relationships by the condition, and the disruption of the child's schooling. It must be carefully planned, and a constant watch kept for adverse effects. These can include difficulty in sleeping, poor appetite, loss of weight and tics. Drugs alone, however, are unlikely to help, and attention must first be given to the situation at home and at school, and how this can be improved. Treatment is often successful, and it can be a rewarding condition to treat. PMID- 10692739 TI - Unstable angina: a review and practical guide to management. AB - The syndrome of increasing angina leading to myocardial infarction was first recognised in the 1930s and the term unstable angina coined in 1971 by Fowler. Since then the importance of acute coronary syndromes as a presentation of ischaemic heart disease has been fully recognised. Recently several new pharmacological agents have been developed for the treatment of these syndromes. We aim to review the current treatments available for unstable angina and give a practical guide to its management. PMID- 10692740 TI - New therapeutic approaches for rheumatoid arthritis. AB - Better understanding of the immunopathogenesis of rheumatoid arthritis over the past few decades has promoted the innovation of new therapeutic approaches targeting the disease more specifically. In addition, refinements in the non steroidal anti-inflammatory drugs (NSAID) may offer advantages for patients with RA. This brief review reports and analyses some of the important aspects of these new therapies available for treatment of RA and in which situations you might consider each. Before using any of these agents, physicians should become thoroughly familiar with the package inserts. PMID- 10692741 TI - Nutritional factors in osteoporosis. AB - Nutritional factors are important but correctable factors in the pathogenesis of osteoporosis. The rate of bone loss in the elderly can be reduced and the peak bone mass in the young can possibly be increased by dietary manipulation, thereby reducing the risk of fracture. Dietary manipulation likely to be of benefit are increased calcium intake, increased vitamin D intake, moderate reduction in intake of salt protein, caffeine and phosphate and increased intake of potassium and magnesium. The possible mechanisms by which these dietary factors influence bone metabolism are discussed. PMID- 10692742 TI - Alcohol, drugs and stigma. AB - Alcohol and drug-related problems are common in clinical populations but are often stigmatized by the public and the professions. These attitudes may interfere with patients obtaining effective help. The reasons for these barriers are explored, including the views that these problems are self-inflicted--and therefore less worthy of help--and the stereotypes and resistances that abound in working with those who have become addicted. Training and public education may go some way to improve this situation. PMID- 10692743 TI - Naratriptan. AB - The new 5-HT1B/1D agonist naratriptan, introduced in many countries in 1997 and 1998 for the acute treatment of migraine, was designed to complement the sumatriptan portfolio of offerings (including the injection, the tablets, the nasal spray, and in some countries, the suppository) by offering patients excellent tolerability and a sustained duration of action. Clinical studies on naratriptan, including more than 4000 patients treating more than 15,000 migraine attacks, show that naratriptan tablets 2.5 mg are distinguished from other 5 HT1B/1D agonists for migraine on the basis of their excellent tolerability profile, which does not differ from that of placebo. In addition to its tolerability, naratriptan tablets 2.5 mg possess a long duration of action with a low incidence of headache recurrence (17-28% in phase II and III clinical trials). With its tolerability profile and long duration of action, naratriptan tablets 2.5 mg may be particularly appropriate as a single-dose alternative to NSAIDs and analgesics, which often are not effective in migraine but are used because of tolerability considerations. PMID- 10692744 TI - Acetazolamide-induced Gerstmann syndrome. AB - Acute confusion induced by acetazolamide is a well known adverse drug reaction in patients with renal impairment. We report a case of acetazolamide-induced Gerstmann syndrome in a patient with normal renal function, to highlight predisposing factors that are frequently overlooked. PMID- 10692745 TI - A rare case of cutaneous kaposiform haemangioendothelioma. AB - Kaposiform haemangioendothelioma is a rare vascular neoplasm with a wide anatomical distribution. We describe an unusual case arising in the post auricular skin of a male infant overlying a ventriculoperitoneal shunt. PMID- 10692746 TI - Oral gastrografin in neonates: a note of caution. AB - Hyperosmolar feeds are known to increase gastrointestinal permeability, predisposing to absorption of toxins. They are also associated with necrotising enterocolitis (NEC) in neonates. A case of a neonate with suspected NEC who died following Gram-negative septicaemia possibly related to oral gastrografin is reported. Hyperosmolarity of gastrografin may have caused complete loss of mucosal integrity in the compromised bowel leading to Gram-negative septicaemia. PMID- 10692747 TI - Fever, rash and worsening of asthma in response to intravenous hydrocortisone. AB - We describe a case of a hypersensitivity reaction to intravenous hydrocortisone in a 64-year-old female with asthma, and briefly review other cases. PMID- 10692748 TI - Lipid-lowering drugs: do the differences matter? PMID- 10692749 TI - IIb/IIIa or not IIb/IIIa? That is the question. PMID- 10692750 TI - Cefaclor af versus amoxycillin/clavulanate in acute bacterial exacerbations of chronic bronchitis: a randomised multicentre study. AB - Cefaclor and amoxycillin/clavulanate are active against Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis and Staphylococcus aureus- pathogens commonly associated with acute exacerbations of chronic bronchitis (AECB). This randomised, parallel-group, single-blind, multicentre study investigated the comparative efficacy and safety of 7-day treatment regimens of cefaclor AF (750 mg b.d. [n = 73]) and amoxycillin/clavulanate (875/125 mg b.d. [n = 72]) in AECB. A favourable clinical response was obtained in 95.9% of patients on cefaclor AF and 97.2% of patients on amoxycillin/clavulanate. There were no statistically significant differences between the groups for improvement in clinical response measured by pulmonary peak expiratory flow (PPEF), or for common symptoms associated with AECB. Both agents were well tolerated, with no statistically significant differences in overall safety; however, nausea and vomiting, and abdominal pain, the most frequently occurring adverse events in the amoxycillin/clavulanate group, were not reported in the cefaclor group. In conclusion, cefaclor AF and amoxycillin/clavulanate have similar efficacy and safety profiles in the treatment of AECB. PMID- 10692751 TI - The effect of valsartan and captopril on lipid parameters in patients with type II diabetes mellitus and nephropathy. AB - The study compared valsartan 80 mg or 160 mg o.d. with captopril 25 mg t.i.d. or placebo on plasma lipids in normotensive and treated hypertensive patients with type II diabetes and microalbuminuria. One hundred and twenty-two adult outpatients were randomised to receive either valsartan 80 mg or 160 mg, captopril 25 mg or placebo for 360 days. Changes from baseline to endpoint in plasma lipid parameters were measured. The primary criterion for tolerability was the incidence of adverse events. All treatment groups showed minor changes in lipid parameters. Triglyceride increased by 2.7% (valsartan 160 mg) to 9.1% (placebo). Total cholesterol decreased under valsartan 80 mg, while other groups showed increases of up to 0.031 mmol/l. Decreases in total cholesterol (p = 0.018), apolipoprotein B (p = 0.042) and apolipoprotein A1 (p = 0.025), were significant for the comparison of 80 mg valsartan and captopril. Valsartan 80 mg or 160 mg o.d. does not cause deleterious changes in the diabetic lipid profile and, unlike captopril, is not associated with dry cough. PMID- 10692752 TI - A dose-defining study of sumatriptan suppositories in the acute treatment of migraine. AB - In this dose-ranging, randomised, multinational, multicentre, double-blind, placebo-controlled, parallel group study, 431 patients treated a single migraine attack with study medication: sumatriptan suppository 6 mg, 12.5 mg, 25 mg, 50 mg, 100 mg, or placebo. Patients were treated in the clinic with a single dose in suppository form. All doses of sumatriptan, except 6 mg, were significantly better than placebo (p < 0.004) and achieved similar rates of headache relief within two hours of dosing. The highest response rate was in the 25 mg group (72%) compared with placebo (37%) (p < 0.001). Fewer patients required rescue medication in the active groups (1% 100 mg to 13% 6 mg) compared with placebo (17%), and more patients were able to work and function normally two hours after dosing (41%, 100 mg; 20%, placebo). The overall incidence of adverse events was similar in the placebo, 6 mg and 12.5 mg groups (14-17%) but higher in the 25 mg, 50 mg and 100 mg groups (25%, 32% and 29% respectively). Analysis of plasma sumatriptan levels indicated rapid rectal absorption for all doses (median tmax = 1.0 hr). It is concluded that sumatriptan, in doses above 6 mg, is an effective and well tolerated treatment for acute migraine. From this study doses of 12.5 mg and 25 mg sumatriptan were identified as having the best efficacy/safety profile and were evaluated further. PMID- 10692753 TI - Rational prescribing: practice audit and drug switch in dyspepsia management. AB - Growing demands on limited healthcare resources and budgets have led to an increased focus on the costs associated with the purchase of drugs. Consequently, approaches to treatment for various disease states are now dictated not only by issues such as best medical benefits, but also by the cost of the drug that is used. As such, drug-switching strategies have become increasingly common practice where, although the clinical benefits offered by the various medications for a specific condition may be similar, the cost differentials are notable. Application of this switch procedure has recently been assessed in the context of the management of acid-related gastrointestinal disorders and has demonstrated that switching patients to therapy with the proton pump inhibitor lansoprazole from H2-receptor antagonists or other proton pump inhibitors not only offers therapeutic advantages but also has important financial implications in general practice with regard to cost savings. PMID- 10692754 TI - Dissolution of thrombus formed during direct coronary angioplasty with a single 10 mg intracoronary bolus dose of abciximab. AB - Direct coronary angioplasty with stent implantation is an effective treatment for acute myocardial infarction. The use of adjunctive abciximab, a platelet glycoprotein IIb/IIIa receptor antagonist is expensive. We report on three cases of direct coronary angioplasty complicated by extensive thrombus formation that were successfully treated with attenuated dosing of abciximab via the intracoronary route. All patients presented with acute myocardial infarction complicated by cardiogenic shock or eminent cardiogenic shock. Abciximab was administered after balloon dilatation when extensive thrombus formation was noted and persisted despite repeated inflations. In all three patients a single 10 mg vial of intracoronary abciximab was administered, resulting in complete dissolution of thrombus, allowing successful deployment of stents. Thus, a single 10 mg intracoronary bolus dose of abciximab may be sufficient to achieve high local concentrations of antiplatelet activity. This facilitates thrombus dissolution and allows the safe deployment of a stent to normalise intracoronary rheology. PMID- 10692755 TI - Comparison of therapy with simvastatin 80 mg and atorvastatin 80 mg in patients with familial hypercholesterolaemia. AB - This study compared the efficacy of simvastatin 80 mg and atorvastatin 80 mg in the treatment of 26 patients with familial hypercholesterolaemia over 12 weeks using an open crossover trial format. Both, similarly, reduced LDL by 47 +/- 13% and 43 +/- 16% and median triglycerides by 22% and 27% respectively. However, atorvastatin reduced HDL by 2 +/- 24% compared with 8 +/- 30% increase with simvastatin (p = 0.05) affecting the LDL:HDL ratio achieved (4.478 +/- 1.56 vs 3.74 +/- 0.93, p = 0.001). Atorvastatin raised median fibrinogen by 15% compared with a non-significant 5% increase with simvastatin (p = 0.05). Simvastatin reduced lipoprotein (a) by a median 20% compared with baseline (p = 0.05) compared with 5% for atorvastatin. Side-effects, mostly gastrointestinal, were seen in four patients (16%) with atorvastatin compared with one case of myalgia with simvastatin (4%). We conclude both drugs are equally effective in LDL reduction but that simvastatin is superior in raising HDL and causes fewer side effects. These results require confirmation in larger studies. PMID- 10692756 TI - Pivmecillinam for the treatment of acute uncomplicated urinary infection. AB - Pivmecillinam is a beta-lactam antimicrobial marketed almost two decades ago. It has been used widely for the treatment of acute cystitis in selected areas of the world, particularly in Scandinavia. With increasing resistance of community Escherichia coli isolates to trimethoprim and trimethoprim/sulphamethoxazole, as previously observed for ampicillin and sulphonamides, reassessment of empiric antimicrobial regimens for acute uncomplicated urinary infection is necessary. Thus, it is timely to revisit the role of pivmecillinam for the treatment of acute cystitis. Clinical studies document the efficacy of this antimicrobial with short course therapy for acute cystitis, and clinical practice in countries where it has been used for many years confirms its efficacy and tolerability. If this agent were more widely used for empiric treatment for acute cystitis, use of antimicrobials such as the quinolones might be avoided. Further trials to define the comparative efficacy of pivmecillinam with other antimicrobials, and further studies of community resistance in E. coli isolates to this agent are needed. PMID- 10692757 TI - Glycoprotein IIb/IIIa antagonists: do they have a role in the management of unstable angina? AB - Plaque rupture, platelet aggregation and thrombosis have central roles in the pathogenesis of acute coronary syndromes. Despite several trials showing the benefit of aspirin and heparin in patients presenting with unstable angina and acute myocardial infarction, these patients are still at risk. This has prompted the development and evaluation of several new therapeutic agents including low molecular weight heparin, new antiplatelet drugs (e.g. ticlopidine and clopidogrel), direct thrombin inhibitors, and intravenous and oral glycoprotein IIb/IIIa antagonists. The IIb/IIIa receptor is the final common pathway involved in platelet aggregation. Thus, whatever the stimulus for platelet activation, subsequent aggregation is mediated by the IIb/IIIa receptor binding fibrinogen. A variety of antibody, peptide and non-peptide compounds that block the IIb/IIIa receptor have been developed, and several studies have investigated the role of these agents in patients with acute coronary syndromes both within and outside the setting of percutaneous intervention. This article summarises the studies to date using IIb/IIIa antagonists, and discusses their role in patients with non-ST segment elevation acute coronary syndromes. PMID- 10692758 TI - Parkinson's disease surgery: advances and future strategies. AB - The recent resurgence of surgical treatment of Parkinson's disease has generated enormous interest in the scientific community and with the general public. Both ablation (pallidotomy, thalamotomy), deep brain stimulation (thalamus, pallidum, subthalamus), and neural transplantation (human, porcine) have been demonstrated to be effective for specific subsets of Parkinson's disease patients. Future studies need to define better the ideal target sites and lesion volume of ablation and stimulation procedures, and optimising graft survival in neural transplantation. Numerous potential surgical interventions are currently being evaluated. PMID- 10692759 TI - The original sin of madness--or how psychiatrists can stigmatize their patients. AB - A stigma is a mark of infamy or disgrace. People who are stigmatized are subject to abuse and social exclusion. Negative attitudes towards people with mental illness are attributed to stigma. In the literature, stigma is regarded as something that is attached to a person--like a badge or label. This is undermined by two facts. Firstly, it is the person who is regarded as disgraceful, not the illness or diagnosis, and secondly, the diagnosis of mental illness is itself evaluative. Mental illness is by definition bad, so to be diagnosed as mentally ill is to be defined as bad (or somehow wrong). The stigmatization of the mentally ill will not stop until this negative evaluation is removed from diagnosis. PMID- 10692760 TI - Calcium dobesilate: pharmacological profile related to its use in diabetic retinopathy. AB - Calcium dobesilate (Doxium) is used in the treatment of diabetic retinopathy. Clinical studies show a slowdown of the progression of the disease after long term oral treatment. The main action of the drug is related to a reduction of microvascular permeability as measured by different parameters and methods (vitreous fluorophotometry, retinal haemorrhages, skin capillary resistance, blood albumin leakage, blood viscosity) leading to improved visual acuity. The pharmacological activity may be explained in part by the antioxidant properties of calcium dobesilate and its action on endothelium through the synthesis of nitric oxide, increasing the endothelium-dependent relaxation. The antioxidant effect was demonstrated in different in vitro and in vivo models by decreasing the peritoneal permeability in rats induced by pro-oxidant substances. Moreover, vascular leakage was also decreased by calcium dobesilate in a reperfusion model in streptozotocin induced diabetic rats after ischaemia of the central artery of the retina. Doxium may also preserve vascular endothelial function by acting directly as antioxidant to protect lipids from peroxidation. PMID- 10692761 TI - Local anaesthetic circumcision in adults. AB - Circumcision is a simple operation often performed under general anaesthesia. In this study, we assessed the suitability of local anaesthesia in 38 adult patients, 79.4% of whom suffered little or no discomfort. Infiltration of local anaesthesia caused moderate pain in 10 patients (29.4%) and severe pain in only one. The circumcision itself caused moderate pain in three patients and severe pain in another three (8.8%); 85% of patients expressed complete satisfaction. PMID- 10692762 TI - Cystic lymphocytic hypophysitis, visual field defects and hypopituitarism. AB - A 45-year-old man presented with anterior pituitary failure, requiring thyroxine, hydrocortisone and androgen replacement. An MRI scan revealed a large cystic pituitary mass and thickening of the pituitary stalk. Over three years, diabetes insipidus and bitemporal hemianopia developed and the cystic mass had enlarged on MR scanning. Transphenoidal resection was performed with normalisation of the visual fields. Histology revealed lymphocytic hypophysitis, which is rare in men. The presentation with cystic enlargement is unique. PMID- 10692763 TI - Iatrogenically revealed myasthenia gravis. AB - Myasthenia gravis is rare. When it presents unusually, making a diagnosis is doubly difficult. We present an unusual case where an 'everyday' medication unfortunately precipitated a myasthenic crisis in an unsuspecting individual. PMID- 10692764 TI - Independence, journals and editors. PMID- 10692765 TI - On the influence of diet and exercise on lipid fractions. PMID- 10692766 TI - One-month mortality rate after liver transplantation for parenchymal cirrhosis: analysis of risk factors in a ten year period. AB - Accurate prediction of short-term survival rate after liver transplantation is one way of selecting recipients and should improve organ allocation. We observed, during the first ten years of our program a striking decline in postoperative mortality with time, a well known observation in Europe as well as in the United States. In 65 adults with parenchymal cirrhosis having received a liver transplant between 1984 and 1994, we examined the possible influence of various preoperative risk factors on one-month mortality rate which was 13.8% in this series. Univariate analysis led to the identification of five significant risk factors: date of transplantation, low serum sodium, previous history of jaundice, ascites and encephalopathy. In the final multivariate analysis however, the date of transplantation emerged as the sole predictive factor of early mortality rate. Therefore, factors such as pretransplantation state of the patient and poor hepatic reserve are counterbalanced by the improvement of surgical skill and other technical aspects, as well as by better perioperative management which have all contributed to the improved results of liver transplantation with time. PMID- 10692767 TI - Histopathological assessment of the prophylactic effect of gingko-biloba extract on intestinal ischemia-reperfusion injury. AB - In this experimental study, the prophylactic effects of Gingko-Biloba Extract (GBE) were examined after experimental ischemia on intestinal wall damage. 50 Wistar-Albino rats (2.5 month old) were gathered and separated into 5 groups (n:10). Group 1 was subjected to a laparotomy (sham-operated group) whereas all other experimental groups were subjected to an occlusion of their superior mesenteric arteries for 30 minutes and a period of 20 minutes reperfusion following occlusion. Group 2 was not given any prophylactic agent during the experiment (untreated control group). GBE was administered in a dosage of 50 mg/kg (i.v.) as a prophylactic agent to Group 3 one hour prior to laparotomy whereas Group 4 was given GBE at 50 mg/kg (i.v.) just before ischemia. Group 5 was given GBE in the same dosage just before reperfusion. Immediately after reperfusion, a biopsy was taken from the ileum (10 cm proximity to ileocaecal valve) for histopathological assessment. A significant prophylactic effect of GBE was observed in Group 5 in which GBE was administered just before reperfusion. PMID- 10692768 TI - Endoscopic therapy of Barrett's oesophagus: critical review. AB - Barrett's oesophagus is known as one of the most important risk factor of oesophageal adenocarcinoma. Because of the increasing incidence of these latter, many endoscopic methods such as argon plasma coagulation, photodynamic therapy or endoscopic mucosal resection are now in evaluation in order to eradicate Barrett's oesophagus or to treat dysplasia and early cancers arising from this metaplasia. The aim of this paper is to comment these techniques and discuss their usefulness. PMID- 10692769 TI - The role of the E-cadherin/catenin complex in gastrointestinal cancer. AB - Cancer is a genetic disease. The unstable genome of cancer cells causes tumour progression through multiple alterations in suppressor and promoter genes, leading to loss of homeostatic and gain of oncogenic functions. Invasion is the critical step in the acquisition of malignancy. It implicates a continuous molecular conversation of the cancer cells with other cells and with the extracellular matrix in which adhesion molecules are crucial. One of these, E cadherin, is discussed in the present review. E-cadherin is a transmembrane glycoprotein that forms a complex with cytoplasmic proteins, termed catenins because they link E-cadherin to the actin cytoskeleton. E-cadherin/catenin mediated intercellular adhesion and communication is mainly homophylic homotypic. There is compelling evidence from experiments in vitro as well as in vivo to accept that the E-cadherin/catenin complex acts as an invasion suppressor. The mechanism of this action is not only through cell-cell adhesion but also through transduction of signals to the cell's motility system. In the replication error positive human colon cancer cell line HCT-8, the alpha E-catenin gene CTNNA1 is an invasion suppressor gene. Here, the transition from the non-invasive to the invasive state was prevented by introduction into the unstable non-invasive cells of either an extra CTNNA1 or a wild type hMSH6 mismatch repair gene. beta-catenin also participates at a complex which comprises the adenomatous polyposis cancer protein APC. In colorectal cancer, mutation of either APC or beta-catenin is oncogenic. Downregulation of the E-cadherin/catenin complex may occur in several ways amongst which are gene mutations, methylation of 5'CpG dinucleotides within the promotor region of E-cadherin, tyrosine phosphorylation of beta-catenin, cell surface expression of proteoglycans sterically hindering E-cadherin and proteolytic release of fragments from the extracellular part of E-cadherin. Upregulation of the E-cadherin/catenin complex has been realized with a series of agents, some of which can be used therapeutically. In most human gastrointestinal cancers the E-cadherin/catenin or related complexes are disturbed and this underscores their pivotal role in the progression of these tumours. Mutations of the E-cadherin gene, including germline mutations, occur in diffuse gastric carcinoma, CpG methylation around the promotor region of E-cadherin in hepatocellular carcinomas and mutations of the APC tumour suppressor gene or in the beta-catenin oncogene in most colorectal cancers. The literature agrees about the disturbance of immunohistochemical patterns of E-cadherin and catenin expression in gastrointestinal cancers. Conflicting opinions do, however, exist about the prognostic value of such immunohistochemical aberrations. We doubt that immunohistochemistry of E-cadherin or catenins add prognostic value to the already used histological grading systems. In our opinion the major benefit from understanding of the E-cadherin/catenin-mediated pathways of invasion will be the development of new anti-invasive treatment strategies. PMID- 10692770 TI - Haemochromatosis and HFE gene. AB - The discovery of the HFE gene has improved classification and diagnosis of iron overload. Most patients with a phenotypic diagnosis of haemochromatosis are homozygote for the C282Y mutation. Among those with other genotypes, only compound heterozygotes, who present the C282Y mutation on one chromosome and the H63D on the other, may present with haemochromatosis, but with a low penetrance and a mild expression. Other patients usually present with another cause of iron overload, such as insulin resistance, alcoholic liver disease or liver cirrhosis. The practical management of haemochromatosis has been greatly modified, since liver biopsy is no more necessary for diagnosis in C282Y homozygotes, and is only needed for exclusion of cirrhosis. Family screening has also greatly benefited from genotyping. PMID- 10692771 TI - Diagnosis and treatment of hepatocellular carcinoma. AB - Several advances have been produced in the diagnosis and treatment of patients with hepatocellular carcinoma. It is possible to diagnose the neoplasm at an early stage when radical treatment options may be applied. The criteria to apply them successfully have been refined and the expected outcome has been improved, but we still lack a useful treatment for the vast majority of patients who are still diagnosed at an advanced stage. Efforts have to be done during the next years to develop such an option (perhaps based on gene manipulation) and until then the management of this tumour will still constitute a challenge for physicians taking care of patients with this neoplasm. PMID- 10692772 TI - NSAID use and abuse in gastroenterology: refractory peptic ulcers. AB - With current antiulcer therapies to eliminate H. pylori infection, non-steroidal antiinflammatory drug use is the main factor involved in resistant peptic ulcers which must be defined as those ulcers that do not heal after 6 (duodenal ulcers) or 8 (gastric ulcers) weeks of treatment with proton pump inhibitors, despite H. pylori eradication. NSAID use (especially aspirin abuse) in patients with resistant ulcers is often surreptitious. Ulcers tend to complicate with stenosis and bleeding, commonly change site, are multicentric and have poorly defined margins. These patients should never undergo surgery unless they develop uncontrolled complications, since ulcer recurrence is the rule. Analgesic abuse and personality disorders might present in some of these patients. Refractory ulcers with no evidence of NSAID use and no evidence of H. pylori infection are rare but not exceptional. Smoking and genetic background seem important factors in patients with this type of ulcers. Idiopathic basal gastric acid hypersecretion might be important in a few patients, but the Zollinguer-Ellison syndrome must be ruled out. PMID- 10692773 TI - Novel nonsteroidal anti-inflammatory drugs. AB - The authors first briefly review how the concept of COX-2 selectivity was brought to light, then tested against the known gastrotoxicity ranking of currently used NSAIDs, from the old classics to the most recent. One truly selective COX-2 agent -celecoxib--is now being marketed in an ever increasing number of countries. So far it seems to keep its main promises, i.e. high--albeit not total--safety regarding gastrointestinal adverse effects, and undisturbed platelet function. Association with warfarin drugs seems to raise no problems, but one should still be wary of possible renal side-effects. Efficacy, at least as assessed in osteoarthritis and rheumatoid patients, appears satisfactory. However, treatment of intense inflammatory crises, such as gout or ankylosing spondylitis, has not been assessed, as yet. Another COX-2 agent--rofecoxib--is on the brink of being released. Its even more potent COX-2 selectivity raises new issues. What about some COX-1 activity that several authors detected in rheumatic synovitis? On the other hand, in particular circumstances, organs such as the stomach, the kidney and small blood vessels, seem to have their homeostasis partly controlled by COX 2 mechanisms also. These questions should be answered soon, whilst clinical experience with the COX-2 agent builds up. PMID- 10692774 TI - Non-steroidal antiinflammatory drugs and the intestine. PMID- 10692775 TI - Hepatitis C recurrence after liver transplantation. AB - Hepatitis C-related cirrhosis is the major indication for liver transplantation (LT). This disease recurs histologically in nearly all the HCV-infected patients during the first postoperative year. Chronic hepatitis C evolves to cirrhosis in 20% of the cases within 5 years after LT. However, the 5-year survival for a HCV infected recipient is still comparable to that of a patient grafted for another indication; it will become worse later. High viremia after LT is associated with a more severe liver recurrent disease. The influence of viral genotype remains controversial. The impact of the type of immunosuppression on HCV recurrence is unclear. Steroids, that increase viremia, might have a deleterious effect on the outcome of chronic HCV-disease after LT. Antiviral combined therapy (Interferon + Ribavirin) soon after transplantation, before disease recurrence, is probably the best treatment at the present time; this remains still unproven. Retransplantation for HCV recurrent cirrhosis allows a 60% survival at 1 year. PMID- 10692776 TI - Multimodality treatment of cancer of the digestive tube: the standard in 1998. Colorectal cancer. PMID- 10692777 TI - Treatment of inflammatory bowel disease with azathioprine: how to use it in 1999. AB - Azathioprine and 6-mercaptopurine are effective drugs in the management of steroid dependent and chronic active inflammatory bowel diseases. They are also well tolerated on the long term. However, their use is still hampered by some drawbacks including delay before efficacy, 20-35% of non responders, relapse at withdrawal of the drugs, possible bone marrow toxicity and other side effects. During the last few years, these drawbacks have been challenged by important studies showing that a better knowledge of the metabolism of these drugs may help to improve their use. PMID- 10692778 TI - New drugs in chronic hepatitis B. PMID- 10692779 TI - A rare cause of biliary pain in Belgium. AB - Ascaris lumbricoides is the most frequent human helminthic parasite. Usually human ascariasis is poorly symptomatic but complications can arise due to worm migration. Erratic worm migration into the biliary tree is a rare but threatening condition regarding the associated complications: cholecystitis, pancreatitis, obstruction of bile ducts, liver abcesses and recurrent pyogenic cholangitis. We describe a case of a young belgian women suffering from recurrent biliary colics over a period of eight months with repeated normal ultrasound findings. ERCP proved being the only effective diagnostic procedure for a living biliary worm, which was successfully removed with a balloon catheter. PMID- 10692780 TI - Diffuse liver angiosarcoma and cerebral cavernous angiomas in a young patient. AB - A case of a thirty-nine year old woman with cerebral cavernous angiomas who developed anaemia and thrombocytopenia secondary to diffuse liver angiosarcoma is reported. This unique association of liver angiosarcoma and cerebral cavernous angiomas may suggest that this tumour may potentially develop from benign vascular lesions. Hematologic abnormalities in angiosarcomas are moreover reviewed based on recent literature search. PMID- 10692781 TI - The mesencephalic trigeminal nucleus in the cat. AB - This review is a concise summary of our current knowledge about the MTN neuroanatomy which in turn is necessary to understand the neurochemistry of this nucleus in the cat. In order to solve the puzzle of neurotransmitter related changes in the synaptic and functional organization of the MTN, we provide a comprehensive description of the neurotransmitter content of MTN neurons. Particular emphasis is given to identifying the possible physiological involvement of MTN inputs in the transmission of proprioceptive information at the first synaptic relay. It is shown that under normal circumstances the large MTN neuron subpopulation contains only Glu that is a strong candidate for a major neurotransmitter in this brain region. However, certain small MTN neurons, most likely interneurons, are found to be GABAergic. Furthermore, NOS immunoreactivity can be detected in the caudal as well as the mesencephalic-pontine junction parts of the MTN and this suggests a mediatory role for NO in some aspects of synaptic transmission in the MTN. The divergent neurochemical content of the cells in the nucleus, should it exist, is likely to be linked with different neuronal functions. Remarkably, no immunoreactivity to any of the neuropeptides examined is observed in the cell bodies of MTN neurons and only fibers and their terminals show peptide-immunolabeling. Most of the labeled peptidergic fibers have immunopositive varicosities that form pericellular basket-like arborizations around unlabeled MTN perikarya. It is predicted that under normal conditions the pericellular arborizations can function as an intranuclear key communication medium between immunopositive projections and immunonegative MTN neurons in the proprioceptive information processing. The levels of transmitter substances in MTN neurons may vary in case of marked changes in the environmental conditions. Axotomy-induced alterations include a long-lasting decrease in the content of CaBPs produced in MTN neurons and/or de novo synthesis of GAL, NPY and CGRP, thus implying the interactive nature and a previously unsuspected neurochemical plasticity of MTN neurons. The newly synthesized neuropeptides can enhance neuronal survival and neurite regeneration. Our results support the assumption that a peptide involvement in the proprioceptive function develops mainly in abnormal conditions. Taken together with the existing neuroanatomical and electrophysiological data, the present results give strong evidence for the occurrence of both excitatory (Gluergic) and inhibitory (GABAergic) transmission in the cat MTN. In addition, evidence is also provided that the MTN receives synaptic inputs from peptidergic and catecholaminergic fibers and these possibly play a significant role in the integration and transmission of trigeminal proprioceptive information. These findings have confirmed the existence of a large number of synaptic contacts in the cat MTN with specific morphological features of their boutons and with presumably different neurotransmitter release from the synaptic vesicles. In this way, knowledge of the origin and neurotransmitter nature of the fibers providing the synapses would facilitate the understanding of the important role of MTN neurons responsible for proprioception in this region. PMID- 10692782 TI - Development and regenerative capacity of descending supraspinal pathways in tetrapods: a comparative approach. AB - Throughout tetrapods a basic pattern in the organization of descending supraspinal pathways is present. The most notable difference between nonmammalian tetrapods and mammals is the apparent absence of somatomotor cortical areas giving rise to long descending projections to the spinal cord. The phylogenetic constancy of descending supraspinal pathways, at least of those arising in the brain stem, probably implies a comparable pattern of development, presumably a developmental sequence in the formation of these central motor pathways. For studies on the development of motor systems, anurans such as the clawed toad, Xenopus laevis, chicken embryos, and opossums are very attractive animals. Moreover, in these species as well as in rodents in vitro approaches can be used. In the present survey, current knowledge on the neurogenesis, axonal outgrowth, synaptogenesis, and developmental plasticity of the central motor pathways in tetrapods including the sparse data available for man, is discussed. These data are placed in the perspective of the development of the spinal cord and, where possible, correlated with functional data. Emphasis is on the clawed toad, X. laevis, chicken embryos, and opossum and rodent data. The outgrowth of axons of descending supraspinal pathways can be regarded as the result of a series of distinct processes, which may be expressed in a coordinated program: (1) the outgrowth of axons and selection of pathways to their appropriate destination; (2) dendritic outgrowth and formation of specific dendritic morphology; (3) selection of specific targets and collateralization by axons; (4) elimination of incorrect and redundant synapses, axonal and dendritic branches, and of mismatched neurons; and (5) functional refinement of synaptic connections. Tracer and transmitter immunohistochemistry in Xenopus laevis showed that from the moment cell division stops, an axon is formed followed by dendrites which emerge from the cell body. At the beginning of the cell differentiation phase the production of the cell-specific neuroactive substances takes place. Initial outgrowth is in a specific direction for each class of neuron. It is likely that all descending supraspinal pathways arise in a similar way. In the spinal projections of each of the descending supraspinal pathways three stages can be distinguished: (1) an initial stage of outgrowth to the spinal cord, (2) a short "waiting" period after which collaterals enter the spinal gray matter, and (3) myelination of axons. An "overshoot" of spinal projections is particularly evident for the mammalian corticospinal tract. The pattern of early descending axonal tracts appears to be similar in all vertebrate groups. Early axons lay down an axonal scaffold containing guidance cues that are available to later generated growth cones. Throughout vertebrates including man, the fasciculus longitudinalis medialis (flm) is the first descending pathway to be formed. Interstitiospinal fibers "pioneer" this tract, and are joined by reticulospinal fibers. Vestibulospinal fibers (the medial vestibulospinal tract) follow much later. The lateral vestibulospinal tract takes a separate course through the brain stem. Late-arriving fiber tracts such as the rubrospinal and corticospinal tracts probably have their own mechanism of selecting the appropriate pathway. The formation of the descending supraspinal pathways occurs according to a developmental sequence. In all tetrapods studied, reticulospinal and interstitiospinal fibers reach the spinal cord first, followed by vestibulospinal fibers and, much later, by rubrospinal and, if present, corticospinal projections. A special case is presented by anurans which in fact have two motor systems, a primary, transient motor system and a secondary, definitive motor system. Reticulospinal, interstitiospinal and vestibulospinal fibers innervate the spinal cord very early in development, well before the development of the hindlimbs. Rubrospinal fibers in PMID- 10692783 TI - [Extracorporeal liver support systems]. AB - Acute liver failure is very life-threatening since the conventional medical treatments have little effects on the clinical outcome. Artificial liver support systems based on blood detoxification alone have proven ineffective because they cannot correct the severe biochemical disorders. An effective liver support system should be capable of carrying out essential functions such as 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 with fulminant liver failure 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 support devices. Some of these systems are currently under clinical investigation in the USA and Europe, and the results of the prospective controlled trials will be soon available. PMID- 10692784 TI - [General features and strategy in the diagnosis of prostatic carcinoma]. AB - Prostate cancer (PCa) is the most commonly diagnosed cancer in men as well as the second leading cause of cancer death. Age, family history and race are proved risk factors for developing a PCa. Prostate specific antigen (PSA) in combination with the digital rectal examination (DRE) has proven to be an essential element in early prostate cancer detection. Enthusiasm for using transrectal ultrasound (TRUS) alone to identify early prostate cancer has not been demonstrated with longer follow-up. The major role of TRUS today is to ensure accurate wide-area sampling of prostate tissue in men with PCa suspicion. This is best accomplished by targeted biopsy of TRUS-suspicious lesions and systematic biopsy of areas without hypoechoic lesions. Urologists recommend digital rectal examination and a PSA blood test annually starting at age 50. PMID- 10692785 TI - [Staging and therapeutic options in local prostatic carcinoma]. AB - Since the introduction of prostate specific antigen (PSA) into widespread use in clinical practice for early detection of prostate cancer (PCa), in combination with digital rectal examination (DRE), there has been a marked increase in the incidence of localized, potentially curable, disease coupled with a simultaneous decline in regional and metastatic prostate cancer in the least years. After the diagnosis of adenocarcinoma of the prostate has been histologically confirmed, an accurate assessment of the stage--or extent--of the disease should be made. We will provide a critical assessment of the currently employed PCa staging modalities. Therapeutic options in locally defined PCa are analyzed. Intracapsular PCa (T1a-T2b N0 M0) is preferably managed by radical prostatectomy, "insignificant" cancers may be treated by watchful waiting. The outcome of irradiation is not as predictable as radical surgery. Neoadjuvant treatment with radical prostatectomy in locally advanced PCa is probably not as efficient as believed; in some instances the value of adjuvant treatment is undecided. As it seems irradiation in conjunction with androgen deprivation can equal efficacy of surgery. PMID- 10692786 TI - [Perineal radical prostatectomy]. AB - At the beginning of the 1980's. P. Walsh and other published their works which dealt with the anatomy and technique of radical retropubic prostatectomy (Reiner 1979; Walsh 1982 + 1983). Since then this operative access carried through simultaneously with an iliacal lymphadenectomy became more and popular. The perineal approach become less important, many urologists had no longer the opportunity to learn this operation. With the introduction of the laparoscopic lymphadenectomy there has been a renewed interest in the radical perineal prostatectomy. Because of exact operation-technique with permanent visual control, generally less blood loss, small complication rates and a diminished morbidity the perineal approach represents another example of "minimal invasive surgery". There are an increasing number of patients with localized prostate cancer in whom this is an appropriate treatment option. PMID- 10692787 TI - [Current options in the treatment of urolithiasis]. AB - Until the 1980s, treatment of upper urinary tract often involved extensive surgical procedures. With the introduction of the extracorporeal shock-wave lithotripsy and refinements to endoscopic instruments and surgical techniques, it has now become possible to destroy stones in most cases without open surgery. Modern stone management can be divided into three steps. The first step covers the treatment of acute pain and the drainage of an obstructed and possibly infected kidney. The second step aims to either remove for destroy the stone. The third step is directed towards the prophylaxis of stone recurrence. PMID- 10692788 TI - [Sexual male dysfunction]. AB - Disorders of libido, erection orgasm and ejaculation are the most important reasons for male sexual dysfunction (MSD), because of organic reasons, medical therapies and social stressing factors, MSD is increasingly evaluated more precisely and it is not limited on erectile dysfunction any longer. With the appearance of the first oral acting substance on the market and the presence of media a great pressure arose to the medical doctors to do prescriptions. Therefore, it is very important to have consultation, diagnostics and treatment of the MSD done with elaborated medical criteria. PMID- 10692789 TI - [Arterial supply in the left colonic flexure]. AB - The authors study the behaviour of the middle colic, left colic superior, middle and inferior and the first sigmoidal arteries in the territory of the terminal portion of the transverse colon, the left colonic flexure and the descending colon. The study was carried out on 1200 angiographies of the superior and inferior mesenteric aa. and on 150 anatomical specimens, surgically extirpated in the course of left emicolectomy operations. Contrary to what is believed by most authors, the left flexure is a colonic tract very well supplied by blood while the descending colon results to be poorly supplied, being served only by one artery (the left sup. colic a.) often of limited caliber and with branches (the middle and the inf. left colic aa.) sometimes totally or partially lacking. In this last colonic tract the vascular continuity, represented by the arterial arcades, is often interrupted. The Riolan's arcade, variously shaped, is to be considered a constant vascular structure (only once it was lacking in this study). Sometimes it is doubled by a second more internal arcade which must not be confused with the intermesenteric arcade. In four of the observed cases, the Riolan's arcade resulted strengthened by a second retroperitoneal arcade, derived from a branching of the middle colic a., whose branches of division went to the two colonic flexures and descended along the postero-lateral walls of the ascending and descending colon, often parallel to the regular abdominal branches. Exceptionally the colonic flexure is supplied by the only left colic a., which behaves as a specific artery, by us called "dominant artery". The central branches of the artery go to the flexure while the lateral ones join the branches of the middle colic and the first sigmoidal aa., effecting tenuous connections, surgically unreliable. In this case the arterial continuity of the Riolan's arcade can be considered interrupted, at least for the surgical practice. The intermesenteric arcade, in its three forms (direct, mixed and indirect), was observed in 20% of the cases. The colic marginal a. is considered by the authors a tier of arches formed by the colic aa. The left colonic flexure is also supplied by particular vessels originated from the middle colic and the left colic aa. (angular branches and arcades and bridge-branches) or from the superior mesenteric a. (angular artery of Donati) and from other sources, particularly from the splenic a. These vessels then join the colic "vasa recta" through the phrenocolic ligament and the marginal omental vessels. This research shows that the vascular continuity of the left colon is not a constant element, able to reassure the surgeon, for possible interruptions that may occur in its composition. PMID- 10692790 TI - [Gynecomastia: diagnostic and surgical approach in the treatment of 61 patients]. AB - Gynaecomastia is a disease with a high incidence (approximately 65% of adult males between 15 and 40 Ys.). In this paper the authors present their experience about the medical and surgical treatment comparing different surgical techniques: adenomammectomy, liposuction and liposuction associated with adenomammectomy. 61 patients ageing 17-42 Ys. (54 with bilateral gynaecomastia and 7 with monolateral disease) were, treated in the Dept. of Plastic Surgery of the Niguarda Ca' Granda Hospital in Milan from 1985 up to 1995. 26 patients were treated with adenomammectomy; Suction assisted lipectomy was used alone in 4 cases and associated with adenomammectomy in 34 cases. The authors suggest that the associated method is the most effective, the aesthetic results being excellent with an important reduction of post-operative complications (mostly referred as haematoma, seroma). PMID- 10692791 TI - [Acute biliary pancreatitis. Role of laparoscopy after 30 years of traditional surgery experience]. AB - The authors herein show their own experience in the treatment of acute biliary pancreatitis. Aim of this study is to evaluate the effectiveness and the safety of the "early" laparoscopic approach to the mild to moderate acute biliary pancreatitis. The authors studied sixty cases of laparoscopic cholecystectomy with intraoperative colangiography for acute biliary pancreatitis (M/F 1:1.2; mean age 59.6 yrs, range 29.79). The patients were divided in two groups on the basis of the severity of the pancreatitis, defined through Ranson's score and Balthazar classification. The mortality rate was nil. Intraoperative morbidity rate was 6.6% in the group I (3/45), and 13.3% in the group II (2/15). Postoperative morbidity rate was 6.7% (3/45) in the group I and 40% in the group II (6/15). The authors show an original diagnostic and therapeutic algorithm for the treatment of acute biliary pancreatitis. Early laparoscopic cholecystectomy with I.O.C. is proposed as the gold standard treatment for mild to moderate acute biliary pancreatitis. This approach appears to be effective and safe in their experience. In case of severe acute biliary pancreatitis, further investigations are mandatory to evaluate the role of laparoscopic approach. PMID- 10692792 TI - [Role of curative local excision in rectal cancer]. AB - The aim of this retrospective study is to evaluate the results of local excision (LE) for rectal cancer for curative purposes. From 1969 to December 1997, a total of 456 operations were performed for surgical treatment of rectal carcinoma (262 males and 194 females, mean age 66 years). 20 patients (4.1%) underwent LE, 7 males and 13 females, median age 65 years. Patients were selected for LE if they met the following criteria during preoperative staging: tumors staged as T1-T2, N0, M0, grading G1 or G2, achievable location. As far as the type of LE is concerned, 13 transanal excisions (Francillon technique), 2 Mason, 2 endoscopic excisions and 3 TEM were performed. Among patients who underwent LE there was no operative mortality. 13 tumors were T1 and 7 were T2; pathologic findings included 20 adenocarcinoma, 14 G1 and 6 G2. There was no postoperative specific morbidity, while aspecific morbidity was minimal (5%). There were no local recurrences but 2 patients (10%) had secondary lesions. Five year overall survival following LE was 87.4%. Comparing T1 and T2 tumors after APR and SSR (17 T1 and 42 T2, all adenocarcinoma), operative mortality and specific morbidity were respectively 1.7% (p = 0.55) and 28% (p = 0.007). There were 5 (8.5%) local recurrences (p = 0.17) and 6 (10.2%) metastatic lesions. Five year overall survival was similar to LE (88.3%; p = 0.76). In conclusion the authors stress the importance that IE for rectal carcinoma must be performed only in selected patients provided there is correct preoperative staging. In these cases five year overall survival, local recurrence and operative mortality were similar to APR and SSR, while there was a statistically significative difference following LE in terms of specific morbidity. PMID- 10692793 TI - [Hernia repair with local anesthesia]. AB - The authors report the results of their own personal experience with inguinal and femoral hernioplasties (424 cases) performed under loco-regional anaesthesia. This anaesthetic approach together with the use of prosthetic techniques (tension free and suture-less) represent the "Gold Standard" in the surgical treatment of hernia. The absence of mortality, the remarkable reduction in terms of postoperative complications, days of hospitalization (one day surgery) represent outstanding advantages. The cooperation of the patient and the stress test at the end of the operation are further advantages of the proposed technique. PMID- 10692794 TI - [Retroperitoneal tumors]. AB - The retroperitoneal tumors are seldom met in their several histological expressions. Authors totally considered 27 patients with retroperitoneal tumors and they were observed from 1975 to 1996: 21 of them were primitively cured in this Institute whereas 6 were affected with relapse or metastasis after a surgical approach which had been performed by other structures. Their operability resulted of 100% with an 87% resectability rate. The surgical mortality resulted nil while the morbidity rate was of 44%. The global survival resulted of 60% to 5 years while the disease-free interval was of 35%. The outliving to 5 years resulted of 65% with a 60% disease-free interval only in the patients who were treated in first instance. The surgical therapy represents the only treatment which can modify the clinical history of such neoplasms. Exeresis should be aggressive enough in order to obtain a total extirpation of the mass and grant a right margin of safety of sound tissue. Such radical proceeding should also be pursued in the treatment of relapses and metastases. The therapeutical efficacy is always linked to the precocity the diagnosis. A strict follow-up of the patients who underwent a surgical operation for retroperitoneal neoplasm is therefore necessary. A minority of the instances was only subjected to a complementary radio and/or chemotherapic treatment. Such protections did not significantly modify the outliving. PMID- 10692795 TI - [Transclavicular approach for delivery of intrathoracic giant goiter. An alternative surgical option]. AB - To remove the immerse portion of a cervical goitre it is necessary to treat preventively the cervical thyroid arteries. In most cases it is afterwards it is easy the blunt finger dissection of the mediastinal bulk following the correct cleavage plane and its dislodging in the cervical area. But in very rare instances, according also to the personal experience, remains some difficulty for the passage of a too bulky and hard mediastinal mass through the rigid limits of the upper thoracic outlet, or the immerse struma is too fragile for pulling it by transfixion threads. Therefore, traditionally arises the opportunity of an additional surgical access, through the breastbone or through the thoracic wall, according to the circumstances. Our experience, completely occasional but extremely positive of two of such cases, induces us to advance a proved alternative surgical proposal. When the difficulty of the removal of the immerse portion of the goitre comes only from the incongruence of the immerse volume and the rigid limits of the upper thoracic outlet, our proposal is that to obtain an amplification of the narrow passage breaking the continuity of the clavicle, by its section beneath the periostium near the breastbone and removing this sternal stump from the joint. The result is that of an widening of the upper thoracic outlet, no more rigid, and making easy the transit of the immerse portion from anterior mediastinum so dislodged in the neck. The rationale of this choice is that all is requested in such cases is only to overcome the obstacle of the incongruence among volume and bulk of the immerse portion and the bone limits fixed from the narrow upper thoracic outlet. Both the traditional sternotomy and the thoracotomy seems disproportional for this purpose, moreover with additional problems during the operation. The true advantage of these classical solutions is in treating under direct vision the anomalous arteries of the mediastinal goitre in cases of ectopic localization. But this is not the case of an immerse cervical goitre. It is therefore essential to note that this proposal applies only to the migrated goitre and not to the ectopic ones. The recovery is extremely simple, and both the aesthetics and the static of the scapular joint are not substantially compromised. PMID- 10692796 TI - [Autologous transfusion technique application table]. AB - The authors make a survey on the reasons leading to the application of different methods of autologous transfusion. They underline, incidentally, the important role played by the issues encountered in dealing with Jehovah's Witnesses as well as the discovery and spread of new transfusion transmitted diseases like AIDS and hepatitis C. They explain their experience, from which they have produced a Best Autologous Transfusion Technique Application Table (Scheda di Applicazione Ottimale delle Metodiche di Autotrasfusione, SAOMA), specific for every type, of operation, through the analysis of many parameters (surgeon, anaesthetist, transfusionist, general conditions of the patients, type of surgical operation). Moreover the authors evaluate advantages and disadvantages of the different autologous transfusion methods, including their cost efficiency aspects, and how they can be combined depending on the type of surgical operation. As a conclusion they attribute great importance to SAOMA to minimize homologous blood transfusion risks, even though at times the clinical aspect is made to prevail over the economic one. PMID- 10692797 TI - [Splenic infarction caused by vascular pedicle torsion]. AB - We report a case of a spleen infarction caused by the vascular pedicle torsion. A 25 year-old-man, heterozygous for HbS, presented with severe abdominal pain especially in the left upper quadrant in front and in the back, fever other symptoms related to acute abdomen. First we excluded most common disease (occlusive one and hematologic one) through conform investigation, then we suspected a spleen problem. So we did further investigation with ultrasonography which showed splenomegaly and the spleen looked twisted with its hilum in contact with previous abdominal wall, moreover (here were are as of decreased signal intensity characteristic of splenic infarction under the capsule and some blood in the Douglas pouch. The patient underwent splenectomy urgently. During the intervention we saw a splenomegaly like the ultrasonography showed, moreover there were a long twisted vascular pedicle and many areas of infarctions, some of which had ruptured causing emoperitoneum. The surgical intervention was successful and the clinical spectrum was solved. The splenic infarction might be clinically silent or to represent a surgical emergency. In front a case of acute abdomen, after exclusion of most common etiology, we underline the importance to suspect a spleen suffering, especially vascular one, when (here was no history of trauma. Considering this fact, a simple not invasive examination like ultrasonography is able to confirm this kind of hypothesis and to give soon information to make the surgical choose. PMID- 10692798 TI - [Obstructive jaundice caused by hydatid cyst rupture in main bile duct]. AB - Hydatid hepatic cyst rupture into bile duct is a complication of hydatid disease. The rupture is more frequent in right or left epatic duct and occasionally in common bile duct (7-9%). A 50-year old man came to emergency room owing to jaundice, fever and abdominal pain. TC show an hydatid cyst with daughter's cyst of left liver and dilatation of biliary tree. Laboratory data of significance included an increased of liver function tests (Bilirubin, Alkaline ph., SGOT, SGPT), VES and leukocytosis. The patients was surgically treated, by total pericystectomy, colecystectomy and coledocotomy with lavage o common bile duct; finally we placed one Kehr drainage and two abdominal drainage. After 15 days of postoperative hospitalization patient was discharged. The best treatment of hydatid cyst is total pericystectomy (when possible). An alternative surgical treatment is possible for the presence of communication with biliary tree. ERCP is very important for a correct diagnosis and for a complete surgical treatment. PMID- 10692799 TI - [Gallbladder agenesis in the laparoscopic era: report of a case]. AB - The authors report an observed case and re-examine the incidence of this defect; they underline the possibility of the diagnosis during the fetal age and the association with one o more defects, sometimes incompatible with the life; in the adult, on the contrary, may be evidence of an associated litiasic pathology of the ++epato-choledocus or of the stenosis of the oddian sphincter. On the base of the diagnostic previous experiences, they elaborate the proposal of an algorythm and underline the utility of a laparoscopic diagnosis either in the pre-operative doubious cases or in the intra-operative diagnosed cases. PMID- 10692800 TI - Adenocarcinoma arising from a recurrent fistula-in-ano. AB - Anal fistulas are frequent events which often recur after an inadequate surgical treatment. Nevertheless their evolution into malignant diseases is infrequently observed. The authors report one case of mucinous adenocarcinoma arising out of a recurrent, long-lasting fistula-in-ano. As reported, abdomino-perineal resection combined with radiotherapy can be the choice treatment. The difficulty is to obtain a reliable differential diagnosis. No imaging technique nor histologic examination can establish a definitive reliable diagnosis; nevertheless, as the risk of adenocarcinoma developing from a long-lasting recurrent fistula-in-ano, although small, is real, authors believe that operative exploration and biopsy of recurrent abscesses and fistulas should be recommended. PMID- 10692801 TI - Recruitment and retention of African American elders into community-based research: lessons learned. AB - Factors influencing the recruitment and retention of African Americans into research studies are not well understood. Studies show that their numbers continue to be low in clinical trials and other nursing studies. However, African Americans have disproportionately high incidences of disease, illness, and death, an important reason for their inclusion in ongoing research. Two urban, community based intervention studies with elderly African American participants are used to show different issues and strategies in recruitment and retention. The sample selection and attrition experiences in the studies are examined using 3 theoretical approaches. Six concepts emerge as fundamental to successful recruitment and retention of diverse population groups: (1) historical cognizance; (2) sanctioning; (3) trust-building; (4) mutuality; (5) recognition of heterogeneity; and (6) researcher self-reflection and introspection. PMID- 10692802 TI - Being a case manager for persons with borderline personality disorder: perspectives of community mental health center clinicians. AB - The scope of case management has expanded to include persons with chronic, nonpsychotic disorders, in particular, persons diagnosed with borderline personality disorder. Despite more widespread use, literature about case management for persons with this disorder is limited. To address this gap in knowledge, a study of the day-to-day experiences of case managers who care for persons with borderline personality disorder was conducted. Seventeen community mental health center case managers gave their informed consent to participate in individual, in-depth interviews. The interviews were analyzed using an interpretive phenomenological research approach. The analysis showed a pattern of monitoring self-involvement. The case managers monitored themselves in terms of expressing concern and setting boundaries. These shared practices highlight a central and unique component of being a case manager for persons with borderline personality disorder, that is, the case manager's focus of attention is on self. By focusing on the self, case managers seek to retain control of the nature of the relationship. The author asserts that the matter to be resolved is not to determine whether retaining or relinquishing control is better, but rather, how best to help practitioners maintain a helpful relationship over time with persons who have borderline personality disorder. In an effort to accomplish this goal, questions about current helping practices and suggestions for working collaboratively with persons who have this diagnosis are provided. PMID- 10692803 TI - Psychological distress in non-Hispanic white and Hispanic abused women. AB - Despite the increasing number of studies that have substantiated that women who have been abused are psychologically distressed, existing research has little focus on women from diverse ethnic backgrounds, and variables that may influence the development of psychological distress have yet to be examined. This study was conducted to examine the correlation of psychological distress with abuse and psychosocial factors in a sample of 62 White and 62 Hispanic abused women. A set of measures of posttraumatic stress disorder, depression, and anxiety showed that White women experienced a higher prevalence of psychological distress than Hispanics. Life changes significantly related to the severity of psychological distress, whereas exposure to abuse was not consistently associated with it. Implications for practice and research are discussed. PMID- 10692804 TI - Arriving at readiness to recover emotionally after sexual assault. AB - The purpose of this grounded theory study was to discover behaviors and processes that lead survivors of sexual assault to seek help with emotional recovery. The substantive theory Arriving at Readiness was developed from interviews with 11 women survivors of sexual assault. Eleven categories, each containing several strategies, form the theory. The core variable is arriving at readiness. The findings clarify why survivors often delay obtaining help with emotional recovery. The theory can be used as a tool to make it more likely survivors will be identified and receive help that is responsive to their needs. PMID- 10692805 TI - Evaluating the effectiveness of progressive muscle relaxation in reducing the aggressive behaviors of mentally handicapped patients. AB - This article reports the results of a study on the evaluation of the effectiveness of muscle relaxation training in reducing aggressive behavior in mentally handicapped patients (MHPs). A pretest and posttest study design was used. Findings showed that there was a reduction of 14.7% of aggressive behavior in the subjects after the muscle relaxation training. Muscle relaxation training appeared to be effective in reducing the frequency of some aggressive behaviors. The strengths and limitations of using muscle relaxation training in reducing aggressive behaviors are discussed. PMID- 10692806 TI - [Cervix cancer and pregnancy. Experience of 5 years at the Gynecology-Obstetrics Hospital No. 3, C. M. La Raza from the Mexican Institute of Social Security]. AB - Of the 300 clinical files of pregnant women, 22 were associated with cancer plus pregnancy in 5 years. Of them 7 were excluded in the work. 15 patients with CaCu and pregnancy were studied in the Gineco-Obstetric Hospital No. 3 C.M. La Raza of the IMSS Mexico City in a period since 1st of January 1988 to 31st December of 1992. Early menarchia and sexual life, take an important roll in the mexican women as predisponent factors, multideliver and tobacco too. The colposcopy with manage cervix biopsy is the election method of diagnosis, with 99.5% of confiability. In the 15 patients, the estirpe was epidermoid cancer in all of them. The diagnosis of pregnancy age made in the first and third trimester of gestation. Only in 3 patients (20%) were founded visible lesion, this rename the necessity of make detection of CaCu during the pregnancy, because we will can found the cases in preinvader stages or early invader, that cure highly, like the cases seen in this work. Stages preinvader and early invaders during the pregnancy are highly curatives. We concluded that they have worst prognosis of survival, aggressivity and early relapse in patients with pregnancy stage by stage. The last because in the Historical Literature review about this topic, there was statistical relevance about pregnancy in the women with CaCu, using our work for reference. CaCu does change his prognosis associated with the pregnancy. PMID- 10692807 TI - [Epidemiology of preeclampsia-eclampsia at the General Hospital O'Horan. (Epidemiology of preeclampsia-eclampsia of a sample, in Yucatan)]. AB - The objective was to describe the epidemiology of preeclampsia-eclampsia (P-E) at the Hospital General O'Horan (HGOH) in Merida, Yucatan, Mexico, from 1995 to 1998. Patients with a discharge diagnosis of P-E were included. Their demographic and clinical data were ascertained. To analyze information, descriptive statistics were used. There were 143 patients. Preeclampsia was documented in 41% and eclampsia in 59%. The mean age of the group was 24.4 +/- 7.3 years. It was found that 76% came from rural area. In 79% schooling was no more than elementary education. Seventy five per cent were married. Sexual life began at a mean age of 18.8 +/- 4.3 years. There was no prenatal care in 27% of the cases. Fifty five per cent were primigravida and 43% multigravida. Nuliparity was documented in 52%. Two o more parities were documented in 48%. Complications were seen in 30%. Overall mortality rate was 5%, more frequent eclamptic patients. At the HGOH, P-E was frequently documented in women with both low socioeconomic status and fewer years of schooling. Prenatal care was also irregular. Clinical evolution was satisfactory in most of them, and the mortality rate was low, although it usually occurred in young eclamptic women. PMID- 10692808 TI - [Trends of perinatal mortality at the National Institute of Perinatology]. AB - The purpose of this study is to address the yearly fetal, neonatal, crude death rates observed at the Instituto Nacional de Perinatologia from 1987 through 1997, and the specific death rates for birth weight, gestation age, cause of death, avoidability, and structure and process failures as proxy to quality of medical care. Data come from death certificates following the WHO criteria which includes the maternal medical history, pregnancy follow up, birth attendance, newborn characteristics, autopsy findings, microbiological results, basic cause of death (of both maternal and fetal/neonatal origin), death avoidability, and structure and process issues. The death certificates were analyzed by the Perinatal Mortality Committee and registered into a computerized database. The fetal mortality rate during 1987 was 17.67 per 1000 births, whereas in 1997 it was 21.5 per 1000 births. There was an increasing tendency from 1987 to 1992, with the highest rate being 34.13 during 1992. After 1992 this rate shows a decreasing tendency. The neonatal mortality rate decreased from 42.82 in 1987 to 17.34 per 1000 live births in 1997. The highest rates were observed among the newborns with the lowest birth weights and at the youngest gestational ages. The most frequent cause of death of maternal origin, in both fetal and neonatal deaths, was premature rupture of membranes. As for the most frequent fetal cause of death was antepartum hypoxia, and among neonatal deaths prematurity and immaturity. The percentage of avoidable perinatal deaths has declined dramatically from 27% in 1987 to less than 10% in 1996. Perinatal mortality at the Instituto Nacional de Perinatologia has decreased during the period under assessment, due to the reduction of the neonatal mortality rate. Improving the quality of medical care focusing mainly on process issues will help lowering avoidable mortality rates. PMID- 10692809 TI - [Bilateral renal agenesis (Potter's syndrome) in a girl born to a hyperthyroid mother who received methimazole in early pregnancy]. AB - Bilateral kidney agenesias (Potter syndrome) in a newborn of a hyperthyroid woman receiving methimazole during early pregnancy. This is a clinical case of a hyperthyroid woman that received methimazole during early pregnancy who gave birth to a girl with bilateral kidney agenesis. The initial clinical data was the presence of oligohydramnios detected by an ultrasound (US) at 19 gestational weeks. Another US at the term of the gestation showed anhydramnios, absence of renal silhouettes and bladder, which was corroborated when the girl was born. She died two days after she was born. Sufficient evidence exists that the methimazole administered during the early pregnancy can cause diverse congenital malformations including Potter's syndrome. PMID- 10692810 TI - [Excretion of uric aid, sodium, and potassium in preeclampsia patients and its behavior in acute hyperglycemia-hyperinsulinemia]. AB - The aim of the present research was to compare the uric acid, sodium and potassium excretions among patients with mild preeclampsia and normotensive pregnancy and to determine their behavior towards an acute physiologic state of hyperglycemia-hyperinsulinemia. It was carried out a cuasi-experimental study with parallel group in 25 patients with mild preeclampsia and in 25 patients with normotensive pregnancy all of them in the third trimester of gestation. The intervention consisted in administering an oral load of 50 grams of glucose in order to achieve a physiologic state of hyperglycemia-hyperinsulinemia. The seric levels of glucose, insulin, creatine, uric acid, sodium and potassium were measured, as well as the last four in urine before the oral load (with at least 6 hours fasting) and 60 minutes after the load, besides that, the urinary excretions of solutes were calculated with standard formulas. The urinary excretions of uric acid, sodium and potassium in fasting, and so after the oral glucose load were lower in the group of preeclampsia patients than in the normotensive gestation group. Upon analyzing the influence of a physiologic state of hyperglycemia-hyperinsulinemia, after the oral glucose load on determined solutes and their urinary excretion, we found that there was a significant decrease in the seric potassium level, without modifying its urinary excretion, as much as in the preeclampsia group as in the normotensive group. The seric uric acid and sodium levels diminished in the preeclampsia group and in normotensive group respectively, without modifying their urinary excretion. In conclusion, in the current study the urinary excretion of sodium, potassium and uric acid were lower in the preeclampsia patients than the women with normotensive pregnancy and a physiologic state of hyperglycemia-hyperinsulinemia didn't modify these excretions. PMID- 10692811 TI - Ozone exposure: a case report and discussion. AB - A 45-year-old man working with ozone presents with evidence of sinusitis, mucus membrane irritation, sleep disturbance and shortness of breath. Naturally occurring or manmade, ozone may damage pulmonary alveolar type I cells at significant exposure levels. EPA and OSHA regulate exposure concentrations. Studies show dose responses with exposures. Supporting epidemiological studies are reviewed briefly. Limiting potential for excess exposure is key to prevention. Recognition of ozone as a potential exposure in the Oklahoma workplace is key to symptom management. PMID- 10692813 TI - A curriculum for stimulating the moral imagination. PMID- 10692812 TI - Improving the health of Oklahomans through clinical prevention. Part 1: Counseling to decrease major risk factors. AB - Compared to other states, Oklahomans suffer higher levels of morbidity and mortality from several common conditions--coronary heart disease, chronic lung disease, stroke and injury. Unhealthy personal behaviors contribute significantly to each of these conditions, thus rendering them at least partially preventable by changing those behaviors. Research has shown that many patients will modify unhealthy behaviors as a result of services provided by physicians or staff in their offices, often with briefly delivered messages. In this report we will discuss the most common preventable illnesses suffered by Oklahomans and the risk factors associated with those illnesses. Physicians should make maximum use of their ability to promote healthy behaviors by their patients, with emphasis on the risk factors associated with significant morbidity in the state. They should also focus on those risk factors patients are likely to change following physician counseling, as determined by prevention research and described in the U.S. Preventive Services Task Force document Guide to Clinical Preventive Services. In general, physicians should consistently deliver messages that address tobacco products, alcohol and other drugs, the use of seat belts, and diet and exercise. Also, they should recommend that all women of childbearing age who are capable of becoming pregnant take a multivitamin containing folic acid daily. PMID- 10692814 TI - AMA litigation center pushes for physician rights. PMID- 10692815 TI - The economy of medical knowledge. PMID- 10692816 TI - Palmoplantar keratoderma (Voerner) with composite keratohyalin granules: studies on keratinization parameters and ultrastructures. AB - A case of the Voerner type palmoplantar keratoderma was studied for abnormalities of keratinization parameters. An enzyme and materials used to build the marginal band or cellular envelope of the cornified cell were all abnormally expressed; i.e. transglutaminase I (TGK), loricrin, and involucrin were abnormally immunostained. In the normal controls, their expression was limited to the upper epidermis, mainly in the granular layer. In the lesional skin, they were detected from the suprabasal layer to the lower horny layer. Filaggrin, the protein of the keratohyalin granule, was also expressed more widely than in controls. Ultrastructural abnormalities included a significantly higher frequency of composite keratohyalin granules than controls, early formation of a marginal band in the midepidermis, and, most remarkably, the clumping of tonofilaments causing vacuolization of the cytoplasm of affected keratinocytes. PMID- 10692817 TI - Repeated 5-aminolevulinic acid-based photodynamic therapy following electro curettage for pigmented basal cell carcinoma. AB - 5-Aminolevulinic acid-based photodynamic therapy (ALA-PDT) in the standard manner is ineffective for pigmented basal cell carcinoma (pBCC), because melanin absorbs the photoactivating light interred for protoporphyrin IX. The objective of this study was to assess the therapeutic outcome of pBCCs with repeated ALA-PDT following removal of pigmentation with electro-curettage. After electro curettage, 16 pBCCs were treated with a combination of topical application of 20% ALA in O/W emulsion and topical instillation of 10% ALA solution, followed by photoactivating light. ALA-PDT was performed more than three times. Fourteen of 16 pBCCs showed CR. Two pBCCs showing PR or NR were excised. Repeated ALA-PDT following electro-curettage was effective for pBCC. PMID- 10692818 TI - Giant nevus lipomatosus cutaneus superficials: case report and review of the literature. AB - Nevus lipomatosus superficialis is a rare disorder characterized by a nevoid fatty growth within the papillary and reticular dermis. Lesions more often occur in the sacral, abdominal, or pelvic regions. A 36-year-old Brazilian female with giant nevus lipomatosus is reported. Our case seems to be the biggest reported in the literature. PMID- 10692819 TI - A case of actinic prurigo in Thailand. AB - Actinic prurigo is a separate entity from the polymorphous light eruption that affects American Indians. It has been reported mainly from North and South America, with only few reported cases from Britain or Asia. We report a case of actinic prurigo in a Thai girl who showed cheilitis and pruritic papules on exposed areas for three years. We were able to induce populovesicular lesions by three consecutive irradiations with 100 J/cm2 UVA and 2 minimal erythematous dose of UVB. However, three weeks after irradiation, a prurigo papule developed at the UVB irradiated site. PMID- 10692820 TI - Trichotillomania and trichophagia leading to trichobezoar. AB - A 14-year-old female presented with the complaints of loss of hair, scalp pruritus, and pain in the abdomen. On careful work-up, she was found to have trichotillomania as well as trichophagia. Investigations also revealed a trichobezoar which completely filled the stomach. Hemogram showed moderate hypochromic anemia. Her detailed psychiatric profile showed a few additional features like obsessive hand washing, knuckle cracking, nose picking and body rocking. Her trichobezoar was removed surgically, and she had an uneventful post operative recovery. She is being maintained on fluoxetine and is doing well. The role of a multi-disciplinary approach to trichotillomania patients is highlighted. PMID- 10692821 TI - Angiolymphoid hyperplasia with eosinophilia showing characteristics of Kimura's disease. AB - Angiolymphoid hyperplasia with eosinophilia (ALHE) is a rare benign condition affecting principally the head and neck region of young females. We describe a 42 year-old female patient of ALHE showing the typical changes of endothelial cells and features similar to Kimura's disease in histologic and immunohistochemical findings. PMID- 10692822 TI - A case of linear scleroderma and myasthenia gravis. AB - We report a case of a 28-year-old woman with myasthenia gravis who developed linear scleroderma seven years later. Myasthenia gravis and scleroderma are rarely found in direct association with each other; there are only five such reported cases, all of which were systemic scleroderma patients. Although localized and systemic scleroderma are distinct entities, autoimmunity is believed to be involved in the pathogenesis of both. Myasthenia gravis and scleroderma may occur coincidentally, but an autoimmune predisposition seems to be the more likely underlying cause, as evidenced by an increased incidence of autoantibodies and autoimmune diseases. PMID- 10692823 TI - Subcutaneous cysticercosis involving the eyelid: sonographic diagnosis. AB - A 25-year-old man and a 14-year-old boy presented with neurocutaneous cysticercosis involving the eyelid. Both patients had hundreds of scattered subcutaneous cysticerci. They were arranged in clusters over the sternocleidomastoid muscle in the neck. Such clustering of cysticerci is highly suggestive of central nervous system (CNS) involvement, as both the sternocleidomastoid muscle and the CNS are supplied by the carotid artery and cysticerci travel via the hematogenous route. We used ultrasonography to diagnose subcutaneous cysticercosis, which showed characteristic low reflective cysts and high reflective scolices inside. Although subcutaneous cysticerci are inconsequential, their verification is important in the diagnosis of more severe CNS involvement. They may be confused with other painless swellings such as lymphadenopathies, neurofibromas, and epidermoid cysts. PMID- 10692824 TI - Large annular purpura and paraneoplastic purpura in a patient with Sjogren's syndrome and cervical cancer. AB - We report a 79-year-old female with anaphylactoid purpura on her legs and unusual large annular purpura on the trunk. Histopathological characteristics of leukocytoclastic vasculitis were observed in the upper and middle dermis of both types of skin lesions. She was complicated by Sjogren's syndrome and advanced cervical cancer. The annular purpura spontaneously resolved in a week and did not recur. However, the anaphylactoid purpura relapsed more frequently and spread more widely following the elevation of her serum SCC antigen levels from the onset of purpura until her death. We consider that the characteristic annular configuration was caused by the complication of Sjogren's syndrome and that the recurrent anaphylactoid purpura indicated paraneoplastic vasculitis primarily caused by the tumor specific protein immune complexes. Complication by Sjogren's syndrome many also play a role in the development of allergic vasculitis. PMID- 10692825 TI - Difficulties in treating steroid withdrawal: intermittent ACTH and low dose systemic cyclosporin used to treat a senile erythroderma patient. AB - The tapering or termination of prolonged strong topical and/or systemic corticosteroid application for extensive generalized eczema has adverse effects on the body and thus presents a very difficult situation. The present case is that of a 68-year-old man with erythroderma following eczema, whose steroid withdrawal was successfully treated with intermittent ACTH and low dose systemic cyclosporin administration over a period of one year. PMID- 10692826 TI - Pityrosporum folliculitis: renal transplantation case report. AB - Pityrosporum folliculitis is caused by the fungus Pityrosporum ovale. It is characterized by the presence of pruriginous follicular papulae and papulae pustules in face, upper part of the trunk, and upper limb root. It is more prevailing in places with hot and humid climates. Its incidence can be associated with either immunosuppressive or chemotherapy states secondary to pathologies. We report herewith a case of pityrosporum folliculitis in a patient who had previously underwent kidney transplantation and these result of the itraconazol therapeutics given. PMID- 10692828 TI - Acquired vulvar smooth muscle hamartoma: a case report and review of the literature. AB - A 36-year-old Korean woman had had a flesh-colored, indurated plaque with pruritus on the labium majora for five years. The lesion was not found in association with hyperpigmented or hypertrichotic patches. Results of biopsy specimens showed an excess of haphazardly oriented smooth-muscle bundles in the mid to lower dermis with an unremarkable overlying epidermis. Our diagnosis was an acquired smooth-muscle hamartoma in the vulva. Although there have been previously reported cases of acquired smooth-muscle hamartoma, this was the first reported case in the vulva. We also describe the characteristics differing between our case and the six previously reported ones. PMID- 10692827 TI - Malignant hidroacanthoma simplex: a case report. AB - Hidroacanthoma simplex is a benign tumor of the skin originating from or showing differentiation to the sweat glands. It grossly resembles seborrheic keratosis of Bowen's disease and histologically shows intraepidermal focal growth of epithelial cells. Malignant transformation of this tumor is rare. We report a case of pigmented hidroacanthoma with malignant transformation in a 67-year-old woman. There was a 20-year history of a skin lesion on the right thigh, which first appeared as a small verrucous papule, progressed to a dark-brown colored patch, and then to a pigmented plaque. Histologically, the primary tumor was composed of small squamoid cells with marked cellular atypia. Most of the tumor cells were located in the epidermis. Immunohistochemically, the cytoplasm of some tumor cells showed a positive reaction for epithelial membrane antigen, but not for either carcino-embryonic antigen or the S-100 protein. PMID- 10692829 TI - Cultured Paget cells derived from the involved skin of a patient with extramammary Paget's disease had an extended life span. PMID- 10692830 TI - Melasma in men: a hormonal profile. AB - Melasma in men is much less common than in women. In the present communication, we evaluated circulating levels of LH, FSH, and testosterone in 15 men with idiopathic melasma. When compared with eleven age matched control men, the circulating LH was significantly higher and testosterone was markedly low in the melasmic men. We conclude that male melasma involves subtle testicular resistance. PMID- 10692831 TI - Antipruritic effect of macrolide antibiotics. PMID- 10692832 TI - Molluscum contagiosum appearing as a solitary lesion on the eyelid. PMID- 10692833 TI - Clinical findings in Japanese children with obstructive sleep apnea syndrome: focus on dental findings. AB - We evaluated clinical findings including those on dentistry and in the oral cavity of children with obstructive sleep apnea syndrome (OSA). This study examined twenty-seven OSA children, who were advised by otolaryngologists to be admitted for closer examination and showed an apnea index (AI) of 5 or more on polysomnographs. Their clinical history was obtained from their mothers, and oral findings were also evaluated. The patient consisted of 15 males (56%) and 12 females (44%). The mean body mass index (BMI) was 16.0 +/- 3.0. Of the clinical findings related to sleeping and the duration of sleeping, snoring was the most frequently observed finding (100%). The mean duration of sleep, calculated from the time they went to bed (9.2 +/- 0.8 p.m.) and the time they got up (7.1 +/- 0.8 a.m.), was 9.9 +/- 1.0 hours. Of the clinical findings obtained during the daytime, hyponasal speech was the most frequently observed finding (74%). In terms of dentistry, oral breathing was the most frequently observed finding (89%). The mean duration of meals was 31.7 +/- 13.8 minutes. Results of oral examination revealed that Hellman's dental age was most frequently IIA. According to the standardized grading classification, grade I was observed in 7%, II in 63%, and grade III in 30% of subjects. PMID- 10692834 TI - Role of T cell subset on the immunosuppression induced by Actinobacillus actinomycetemcomitans. AB - Purified splenic T cells from C3H/HeN mice primed with sonic extract (SE) from Actinobacillus actinomycetemcomitans were adoptively transferred to syngeneic mice with sheep red blood cell (SRBC). The transfer of splenic T cells from mice, primed with SE for 8 days, resulted in the dose-dependent inhibition of IgM anti SRBC plaque forming cells (PFC) compared with normal and BSA-primed splenic T cells. Furthermore, the transfer of cells from mice primed with 200 micrograms of SE for 8 days to syngeneic mice caused the highest inhibition. Immunosuppression did not depend on the B cell population in spleen from donor mice primed with SE. Splenic T cells from SE-treated mice could suppress the T cell-dependent proliferative responses of co-cultured normal spleen cells in vitro. Analysis of T cell subsets of spleen cells from mice treated with immunosuppressive factor (ISF) showed that the suppressor cell is susceptible to treatment with anti-CD8 and complement (C). SE-sensitized suppressor T cells also suppressed the secondary IgG anti-SRBC-PFC response after immunization with SRBC in vivo depending on sensitized periods induced by ISF. Treatment of T cells from mice which primed with ISF for 8 days, with goat anti-mouse CD8 antibody and C abrogated their suppressive effects, and secondary IgG response occurred. These results indicate that the adoptive transfer of SE-induced T cells, which increased suppressor function, caused the perfect blocking of the immunoresponse, allowing promotion of secondary infection. PMID- 10692835 TI - Characteristics of proportional analysis for soft tissue facial profile: epidemiological possibilities of measurement item reduction. AB - Lundstrom et al. proposed a proportional analysis system for the soft tissue facial profile in the natural head position. To use this method for further epidemiological investigation and to interpret the characteristics of this analysis, each measurement (index) was identified in comparison to the other indices using cluster and factor analyses. Facial profiles of 111 (mean age: 22.9 years) Japanese males were measured and 11 indices (8 horizontal, 2 vertical and 1 horizontal/vertical) were calculated. Almost all internal co-relationships between each index were statistically significant (p < 0.05, 0.01). Variable cluster analysis classified indices into four major clusters and clarified the attributes of the 11 indices. The first cluster was index No. 1, 2, 3 and 7. The second cluster was index No. 6. The 3rd cluster was index No. 4, 5, 11, 8 and 10. The 4th cluster was index No. 9. These clusters are thought as vertical facial balance, upper and lower jaw relation or horizontal/vertical balance, chin morphology, and horizontal facial balance. From factor analysis, three factor axes that explained the characteristics of 11 indices were found (accumulated contribution rate: 76.5%). The heaviest loading factor was index No. 1,2 (0.95) on axis I, 5 (0.83) on axis II and 6 (0.78) on axis III. Therefore, axis I, axis II and axis III are thought to be based on the position of the soft tissue Nasion, SLI and Pogonion, respectively. Common indices which are included in both analyses are thought to be valid as a clue to reduce the number of measurement parameters. PMID- 10692836 TI - Co-aggregation as a virulent factor of Streptococcus sanguis isolated from infective endocarditis. AB - The pathogenicity of strains of the Streptococcus sanguis group, isolated from infective endcarditis, was studied by measuring the development of subcutaneous abscesses in mice after infection with S. sanguis and Actinomyces viscosus either singly or as co-aggregated pairs. The pathogenicity of the co-aggregates was also examined in various viable combinations of the two bacterial species. More abscesses were formed by A. viscosus than the S. sanguis group including clinical isolates. Abscess formation by co-aggregates of combinations of each isolate and A. viscosus produced a higher percentage of abscess formation than those caused by infection with a pure suspension of A. viscosus or tested streptococci. Co aggregated cells were more resistant to phagocytosis and killing by neutrophils in vivo. These results indicated that S. sanguis group streptococci isolated from infective endocarditis are able to co-aggregate and resist phagocytosis. The ability of co-aggregation of S. sanguis may serve as a survival mechanism in a host defense system and may be linked with virulence of this bacteria. PMID- 10692837 TI - Mandibular-position sensation during sedation by administration of nitrous-oxide (N2O) gas. AB - Our aim was to confirm the influence of N2O gas on mandibular-position sensation. The subjects in this study were eight healthy adults. Each subject was asked to hold the reference stick for five seconds between the central tooth in his or her upper and lower jaws. Then, the reference stick was replaced by the test sticks with different thickness, each of which the subjects were again asked to hold at the same position for five seconds. The subjects were instructed to determine, based on judgement of the interincisal distance, whether the thickness of each test stick was larger or smaller than the reference stick. A series of trials was administered to each subject using all eight sizes of test sticks. We compared the ability of the subjects to discriminate mandibular position both before and after the application of a vibrating stimulus to the masticatory muscles, and before and during the administration of N2O gas. Discrimination ability was significantly decreased after the application of the vibrating stimulus. However, during the administration of N2O gas, no significant difference in discrimination ability was observed between before and after the vibrating stimulus. The results of this study indicated that N2O gas had an inhibitory effect on gamma-motor neuron activity, which is presumed to be mediated to some extent through the higher central nervous system. This is the case because the gamma-motor neurons are generally activated by vibrating stimuli applied to the muscle causing decreased discrimination ability of mandibular position. Thus, we conclude that mandibular-position sensation was influenced by N2O gas during the administration of N2O gas. PMID- 10692838 TI - A case of an electrical burn in the oral cavity of an adult. AB - Electrical burns in the oral cavity account for 2.2% of all electrical burns and only 0.12% of all burns; thus, the incidence of electrical burns in the oral cavity is relatively low. As this type of injury occurs in the oral cavity when an individual sucks or chews on a live electrical wire, extension cord, plug, or outlet, most cases occur in toddlers or preschool children, and adult cases are extremely rare. Here we describe a case of an electrical burn in a 56-year-old man who accidentally bit the electric wire of a cleaner while carrying out repairs. Conservative treatment, without surgery, was performed. Two years after the injury, a slight scar and a small tongue deformity remain, but no functional disturbance has been observed. PMID- 10692839 TI - Oral collision carcinoma: salivary duct carcinoma of minor salivary gland origin and squamous cell carcinoma of the oral mucosa. AB - This paper reports a case of oral collision carcinoma consisting of salivary duct carcinoma of minor salivary gland origin and microinvasive squamous cell carcinoma of the oral mucosa in a 65-year-old Japanese man. This is an exceedingly rare example of collision carcinoma in the oral region. PMID- 10692840 TI - Worsening of pre-existing TMJ dysfunction following sagittal split osteotomy: a study of three cases. AB - When patients seeking treatment for malocclusion also suffer from temporomandibular joint (TMJ) disorders, it is hard to predict the result of simultaneous treatment of both conditions, or to plan for its different goals, because of unpredictable changes in the relationship between the disk, the fossa and the condylar head. Prediction is harder in cases of presurgical TMJ hypomobility, especially those with adhesion in the upper TMJ compartment. Authors differ widely on the likely effect of orthognathic surgery on TMJ disorders. This paper reports three cases in which TMJ disorders worsened after treatment of malocclusion by sagittal split osteotomy. It examines how presurgical diagnosis of TMJ disorders could assist treatment planning in such cases. The results suggest that microbleeding in the upper TMJ compartment during orthognathic surgery, as well as long-term postoperative intermaxillary fixation, carries a risk of creating worse adhesion that adversely affects the outcome for patients. Therefore, preoperative diagnosis of disk position and pathological conditions in the upper TMJ compartment, as well as careful choice of method and term of postoperative fixation, are essential in planning the treatment of malocclusion with sagittal split osteotomy. PMID- 10692841 TI - Control of early ovarian follicular development. AB - Early follicular growth refers to the development of an ovarian follicle from the primordial to early antral phase. In sheep and cows these phases of growth can be classified by the configuration of granulosal cells in the largest cross-section of the follicle as types 1 (primordial), 1a (transitory) 2 (primary), 3 and 4 (preantral) and 5 (early antral). Follicles classified as type 1 may be highly variable within each species with respect to number of granulosal cells and diameter of oocyte. Much of the variation in granulosal cell composition of type 1 follicles may occur at formation and this may account for the variability in granulosal cell composition throughout subsequent stages of growth. There appear to be important differences among species (for example sheep and cattle) in the number and function of granulosal cells relative to the diameter of the oocyte during the initiation of follicular growth. There is evidence that most, if not all, of the growth phases from types 1 to 5 are gonadotrophin-independent and that follicles develop in a hierarchical manner. In sheep, cows and pigs, numerous growth factor, growth factor receptor and gonadotrophin receptor mRNAs and peptides (for example c-kit, stem cell factor, GDF-9, beta B and beta A activin/inhibin subunit, alpha inhibin subunit, follistatin, FGF-2, EGF, EGF-R, TGF beta 1,2 and 3 FSH-R and LH-R) are expressed in a phase of growth (for example types 1-5)-specific and cell-specific manner. However, the roles of many of these factors remain to be determined. PMID- 10692842 TI - Comparative patterns of follicle development and selection in ruminants. AB - Expanding technological capabilities, particularly in ultrasonography and molecular endocrinology, have bridged the gap between form and function of the ovary, and have been a catalyst for intense research activity in this area during the last decade. However, the study of follicular dynamics is still in its infancy in ruminant species other than cattle, and controversy persists regarding the pattern of follicular growth and the existence of follicular dominance. The bovine model of ovarian function is presented as a foundation for concepts surrounding the control of follicular development in ruminants, and to place in context the results of recent studies in sheep, goats, muskoxen, cervids and camelids. This comparative approach is used to determine important generalities that appear to be applicable, as fundamental physiological phenomena, to all ruminant species. Although clear differences in follicular dynamics are evident, differences appear to be specific rather than general, and the following conclusions are consistent with results reported in ruminant species to date: (1) follicles grow in a wave-like fashion; (2) periodic surges in circulating FSH are associated with follicular wave emergence; (3) selection of a dominant follicle involves a decline in FSH and acquisition of LH responsiveness; (4) periodic anovulatory follicular waves continue to emerge until occurrence of an LH surge (that is, at the time of luteolysis during the ovulatory season or during transition from the anovulatory season); (5) within species, there is a positive relationship between the duration of the oestrous cycle and the number of follicular waves; (6) progesterone suppresses LH secretion and growth of the dominant follicle; (7) the duration of the interwave interval is a function of follicular dominance, and is negatively correlated with circulating FSH; (8) follicular dominance in all species is more pronounced during the first and last follicular waves of the oestrous cycle; and (9) pregnancy, the prepubertal period and seasonal anoestrus are characterized by regular, periodic surges in FSH and emergence of anovulatory follicular waves. PMID- 10692843 TI - Molecular mechanisms regulating follicular recruitment and selection. AB - Ovarian follicular growth and development is an integrated process encompassing both extraovarian signals, such as gonadotrophins and metabolic hormones, and intraovarian factors. Follicular development has been classified into gonadotrophin-independent and -dependent phases. In the latter, FSH provides the primary drive for follicular recruitment and LH is required for continued development of follicles to the preovulatory stage. A transient increase in circulating FSH precedes the recruitment of a group of follicles, and these recruited follicles are characterized by expression of mRNAs encoding P450scc and P450arom in granulosal cells. As follicles mature, there is a transfer of dependency from FSH to LH, which may be part of the mechanism(s) involved in selection of follicles for continued growth. Indeed, changes in the pattern of expression of mRNA for gonadotrophin receptors and steroid enzymes within follicular cells appear to be closely linked to changes in peripheral concentrations of gonadotrophins. The mechanism of selection of dominant follicles still requires clarification, but seems to be linked to the timing of mRNA expression encoding LHr and 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) in granulosal cells. Additional intraovarian systems, including the ovarian IGF and activin/inhibin systems, also exert a role. For example, it appears that the development of follicular dominance in cows is associated with the FSH-dependent inhibition of the expression of mRNA encoding insulin-like growth factor binding protein 2 (IGFBP-2) in granulosal cells. In conclusion, the integration of these endocrine signals and intraovarian factors within follicles determines whether follicles continue to develop and become dominant or are diverted into apoptotic pathways leading to atresia. PMID- 10692844 TI - Role of growth hormone in development and maintenance of follicles and corpora lutea. AB - Growth hormone (GH) is a pituitary hormone that affects animal growth, metabolism, lactation, and reproduction. Many of the effects of GH are mediated by insulin-like growth factor I (IGF-I) which is synthesized in liver and ovary in response to GH. Insulin-like growth factor I synergizes with gonadotrophins (LH and FSH) to stimulate growth and differentiation of ovarian cells. There are species differences in the effects of GH in reproductive biology. In most species, ovarian follicles and corpora lutea are potential sites for GH action because the GH receptor is found within granulosal cells as well as corpora lutea. However, growth hormone does not control ovarian IGF-I in all species and, in ruminants, endocrine IGF-I from liver may be the principal mediator of GH action. In cattle, administration of GH increases the number of small antral ovarian follicles but does not increase the number of large antral (dominant) follicles. Growth hormone may antagonize some aspects of dominant follicular function because dominant follicles are shorter-lived in GH-treated cattle. The corpora lutea has increased growth and steroidogenesis in response to GH. Growth hormone-induced steroidogenesis in cultured granulosal and luteal cells depends on IGF-I release after GH treatment. Bovine and ovine granulosal cells do not release IGF-I in response to GH in vitro and, therefore, are less responsive to GH. These results demonstrate that GH is required for normal reproductive function in ruminant as well as nonruminant species. PMID- 10692845 TI - Regulation of follicle waves to maximize fertility in cattle. AB - Cattle have recurrent follicular waves every 7-10 days in most physiological situations; an FSH increase is associated with emergence of the wave and LH pulse frequency determines the fate of the dominant follicle. To control oestrus with hormones it is necessary to ensure that either induced corpus luteum regression or the termination of a progestogen treatment coincides with the selection of the dominant follicle during the wave, to give a precise onset of oestrus and high fertility. The exogenous administration of progesterone or progestagen blocks the normal turnover of the dominant follicle once the corpus luteum regresses. Thus, the effects of duration of dominance of the preovulatory follicle on onset of oestrus and fertility were examined. The variation in onset of oestrus was reduced but occurred 5-9 h later after 4 versus 8 days of dominance; pregnancy rate was also affected with dominance periods of 2-4, 4-8 and > 10 days resulting in 0, 10-15% or 20-50% reduction in pregnancy rates, respectively. The necessity for short duration of dominance of the preovulatory follicle means that to ensure high fertility the follicular wave needs to be regulated when using hormones to control oestrus. Two approaches were examined, namely the use of GnRH or oestradiol at time of progesterone intravaginal releasing device insertion. The effect of 250 micrograms of synthetic GnRH on the fate of an existing follicle wave was to ovulate the dominant follicle (20/20 cows) and a new wave emerged 1.6 +/- 0.3 days later; however, there was no effect of GnRH on the wave if administered before dominant follicle selection. The effect of oestradiol concentrations on suppression of FSH in ovariectomized heifers showed that increasing oestradiol to 10-15 pg ml-1 caused a 37 +/- 6.9% decrease in FSH for 24 h, with a subsequent increase to pretreatment values by 57 +/- 13 h. In cyclic heifers, increasing oestradiol to > 10 pg ml-1 in conjunction with progesterone treatment at emergence of the first wave of the cycle affected the current follicle wave by either preventing dominant follicle selection or decreasing diameter of the dominant follicle, without consistently affecting the interval to new wave emergence. Increase of oestradiol after dominance, however, delayed new wave emergence by 2-5 days. A better understanding of the hormonal control of follicle waves will lead to development of improved hormonal regimens to control oestrus sufficiently to give high pregnancy rates to a single AI without recourse to detection of oestrus. PMID- 10692846 TI - Regulation of GnRH receptor gene expression in sheep and cattle. AB - The GnRH receptor plays a pivotal role in reproduction. This review summarizes current knowledge of the regulation of GnRH receptor gene expression by endocrine factors in sheep and cattle. Expression of the GnRH receptor gene, measured by steady-state amounts of GnRH receptor messenger RNA (mRNA), is maximal during the preovulatory period. The molecular events leading to maximal GnRH receptor gene expression are probably triggered by decreased circulating concentrations of progesterone at luteolysis. Because GnRH is a positive homologous regulator of its own receptor, increased pulsatile GnRH after removal of negative feedback effects of progesterone stimulates expression of the GnRH receptor gene early in the preovulatory period. Oestradiol is also a positive regulator of GnRH receptor gene expression, and increased serum concentrations of oestradiol from developing follicles probably maintain high abundance of GnRH receptor mRNA later in the preovulatory period. Since increased amount of GnRH receptor mRNA precedes maximal numbers of GnRH receptors before the LH surge, increased expression of the GnRH receptor gene is an important mechanism by which maximal sensitivity of gonadotrophs to GnRH is achieved. Future efforts should be directed towards elucidating the molecular mechanisms underlying transcriptional regulation of the GnRH receptor gene in ruminants by endocrine factors. PMID- 10692847 TI - Follicle-stimulating isohormones: regulation and biological significance. AB - Follicle-stimulating hormone (FSH) is a key hormone in the regulation of follicular development. Although the existence of FSH heterogeneity is well established, the physiological significance of this pleomorphism remains unknown. Observed changes in circulating FSH heterogeneity during critical reproductive events such as puberty and reproductive cyclicity suggest that different combinations of FSH isoforms reach the target sites during different physiological states to influence a variety of biological end points such as cellular growth, development, steroidogenesis and protein synthesis. Considering that these FSH isoforms have different physicochemical properties and potential to bind not only their cognate receptors but also structurally related, non-FSH receptors with various affinities, the regulatory implications of FSH heterogeneity in modulating the various FSH-induced functions are enormous. However, assigning functional significance to FSH heterogeneity has been hampered because of (1) difficulties associated with procurement of highly purified, naturally occurring, circulating FSH isoforms; (2) absence of reference standards that contain the entire repertoire of FSH isoforms present in biological fluids; and (3) specificity issues inherent to the detection systems used. If particular FSH isoforms do possess selective biological functions, specific combinations of FSH isoforms could be generated to regulate fertility in farm animals and humans. PMID- 10692848 TI - Endocrine and reproductive responses of male and female cattle to agonists of gonadotrophin-releasing hormone. AB - The pituitary response in cattle to treatment with GnRH agonist has two phases. In the acute phase secretion of LH is increased, while the chronic phase is characterized by a downregulation of GnRH receptors and insensitivity of gonadotrophs to natural sequence GnRH. After long-term treatment with GnRH agonist, cattle do not have pulsatile secretion of LH but maintain basal LH. This is associated with reduced pituitary contents of LH, LH mRNA, FSH and FSH mRNA. Long-term treatment of bulls with GnRH agonist results in an increase in testicular LH receptors and high plasma testosterone. Heifers treated with a GnRH agonist from early in the oestrous cycle develop a larger corpus luteum and secrete more progesterone. Increased steroidogenesis is reflected in increased steroid acute regulatory (StAR) protein and steroidogenic enzymes in the testes and corpus luteum. GnRH agonists have potential as novel strategies for reproductive management in cattle. A GnRH agonist bioimplant was recently used to block the LH surge after FSH stimulation of follicle growth in heifers. Ovulation was induced by injection of LH, and heifers were inseminated relative to the LH injection. This GnRH agonist-LH protocol provides a model for studying the gonadotrophin requirements for follicular growth and oocyte maturation in cattle, and will enable controlled in vivo maturation of oocytes before recovery for in vitro procedures. PMID- 10692849 TI - Control of parturition in ruminants. AB - Parturition is a process which, when set into motion, occurs to completion. This review concerns the control of parturition in ruminants. Parturition is an endocrine event, dependent upon the activation of the fetal hypothalamus pituitary-adrenal (HPA) axis. In sheep and other ruminants, increases in plasma concentrations of cortisol induce the activity of 17-hydroxylase and 17,20 lyase in the placenta, increasing the biosynthesis of oestrogen relative to progesterone. The increase in the so-called E:P ratio increases myometrial activity and culminates in labour and delivery. Much work has been done to identify the mechanism of the endogenous activation of the fetal HPA axis. Recent work suggests that production of prostanoids within the fetal brain influences fetal ACTH secretion, and that induction of prostanoid biosynthesis at the end of gestation might be important in the process of parturition. Oestrogen and androgens, secreted by the placenta at the end of gestation, augment activity of the fetal HPA axis by increasing fetal ACTH secretion and by decreasing negative feedback sensitivity to cortisol. Although significant progress has been made concerning the neuroendocrinology of parturition, many significant questions remain. Is parturition regulated or simply programmed? Is parturition the ultimate result of neuronal maturation within the fetal hypothalamus, or is there a complex interplay between the placenta and fetal hypothalamus? Answers to these and other important questions await further research, but may provide key information which will prove useful in understanding general principles of parturition in many mammalian species. PMID- 10692850 TI - Implications of recent advances in reproductive physiology for reproductive management of goats. AB - The control of reproduction in goats is interesting for technical reasons (synchronization of kiddings, adjustment to forage availability or to economy), and for genetic reasons (identification and dissemination of improved genotypes). The use of short-light rhythms leads to markedly increased production of semen per buck and prevents occurrence of a 'resting' season. Recent identification of a bulbourethral lipase in goat spermatozoa opens new perspectives in sperm preservation. Light plus 'short day' treatments also allow induction of out-of season oestrous cycles and ovulations leading to enhanced fertility. Repeated use of eCG provokes the production of antibodies, delays the timing of ovulation and causes a reduction in fertility after fixed-time artificial insemination. All steps of embryo production, freezing and transfer are now controlled and allow the attainment of satisfactory numbers of kids born per donor female, which are compatible with the development of the technique for exchanging genotypes between countries. In vitro production of embryos allows high development rates to be achieved after in vitro maturation and fertilization of oocytes, and will ensure the production of synchronous populations of one-cell zygotes at the stage required by new biotechnologies. PMID- 10692851 TI - Comparative reproductive function in cervids: implications for management of farm and zoo populations. AB - The cervids represent a complex assemblage of taxa characterized by extreme diversity in morphology, physiology, ecology and geographical distribution. Farmed species (for example red deer and fallow deer) are usually the common larger-bodied, gregarious and monotocous species that express marked reproductive seasonality in their temperate environment. Their commercial importance has facilitated considerable research into reproductive physiology and the development of assisted reproductive technologies (ART). In contrast, the remaining species, including many of tropical origin, show wide diversity in reproductive patterns, have generally received little scientific scrutiny, and include a number of endangered taxa that are reliant on ex situ conservation efforts (such as captive breeding) to ensure their survival. Domestication and ex situ management programmes have been associated with widespread translocation of various cervid species around the world, often placing the animals in environments that are not compatible with their evolved reproductive patterns. For example, the summer calving/lactation pattern of red deer, attuned to northern continental climatic patterns, is frequently misaligned with seasonal changes in feed availability in the Australasian pastoral environment. Similarly, seasonal or aseasonal calving patterns of tropical species translocated to temperate regions are usually associated with increased perinatal mortality of calves born in cool seasons. Conversely, temperate species in tropical zones may exhibit aberrant reproductive patterns in the absence of biologically significant photoperiod fluctuations. ARTs, which presently include artificial insemination, embryo transfer and in vitro embryo production, have potential application to the genetic management and population growth of various cervid species. Although application to some farmed cervid species is widespread, these technologies are rarely directly transferable from farmed to endangered species. Even within species, ART protocols developed successfully for one genotype (i.e. subspecies) may be ineffective in another (for example superovulation of red deer and wapiti). Therefore, application to genetic management of endangered species necessitates prior research into their reproductive patterns. This is often difficult because of the rarity of the animals, a lack of suitable handling facilities for the particular species, and the timid nature of the deer. More recently, however, non-invasive reproductive profiling, based on remote collection and monitoring of excreted steroid metabolites, has facilitated such research. PMID- 10692852 TI - Reproduction in water buffalo: comparative aspects and implications for management. AB - The domestic buffalo occupies an important niche in many ecologically disadvantaged agricultural systems, providing milk, meat and draught power. Although buffalo can adapt to harsh environments and live on low quality forage, their reproductive efficiency is often compromised by such conditions. Climatic stress depresses ovarian cyclicity, oestrous expression and conception rates. Poor nutrition, usually related to seasonal fluctuations in availability and quality of feed, delays puberty and increases the duration of postpartum anoestrus. Management factors such as the system of grazing (free, tethered or none) and sucking by calves (restricted or ad libitum) also modulate reproductive functions. Finally, the skills and capabilities of farmers as well as the quality of support services such as artificial insemination and disease control also influence fertility. The relative importance of these factors vary greatly depending on ecological conditions and production systems. Improvement of reproductive efficiency therefore requires the identification of specific limiting factors under a given situation and the development and field testing of strategies for improvements and interventions that are sustainable with available local resources. The application of modern reproductive technologies in buffaloes requires an appreciation of their biology and reproductive physiology as well as the potentials and limitations under each specific production system. PMID- 10692853 TI - Reproduction in female South American domestic camelids. AB - Alpacas and llamas are induced ovulators. They show marked reproductive seasonality in the Andean region, but under Northern Hemisphere conditions of feeding and management, they are non-seasonal breeders. Puberty is attained when they reach 50% of adult body weight. When they are not exposed to a male, females show successive waves of follicular maturation and atresia. Growth, maintenance and regression of a follicle each require an average of 4 and 6 days in alpacas and llamas, respectively. After sterile mating, progesterone concentrations in blood were increased from day 5, reached maximum concentrations on day 7-8, and declined rapidly at 9-10 days after mating. A fertile mating results in formation of a corpus luteum that remains functional throughout gestation. The duration of gestation is 340-346 days. Almost all fetuses were found to occupy the left uterine horn, even though ovulation occurs from both ovaries with equal frequency. Several methods of pregnancy diagnosis have been described. Mating is recommended within 15-20 days after parturition to obtain good fertility rates and one offspring per year. The factors that contribute to high rates of embryonic mortality are unknown. Reproductive technologies, such as AI, superovulation, embryo transfer and IVF, have not been used very extensively in these species but can be successfully applied. PMID- 10692854 TI - Growth and development of the corpus luteum. AB - The mammalian corpus luteum, which plays a central role in the reproductive process because of its production of hormones such as progesterone, is an exceptionally dynamic organ. Growth and development of the corpus luteum are extremely rapid, and even when the corpus luteum is functionally mature cellular turnover remains high. Associated with this high rate of cell turnover, the mature corpus luteum receives the greatest blood supply per unit tissue of any organ, and also exhibits a relatively high metabolic rate. Central to the growth and development of the corpus luteum, therefore, is luteal vascular growth, which appears to be regulated primarily by the angiogenic growth factors, basic fibroblast growth factor and vascular endothelial growth factor. In addition, the corpus luteum is a complex tissue composed of parenchymal (small and large steroidogenic) and nonparenchymal (for example fibroblasts, vascular smooth muscle, pericytes and endothelial) cells. Recent studies evaluating the expression, location and regulation of gap junctions in the corpus luteum indicate an important role of gap junctional intercellular communication in the coordination of function among these diverse cell types during luteal growth and development. These studies will lead to an improved understanding not only of luteal function but also of tissue growth and development in general. PMID- 10692855 TI - Molecular regulation of luteal progesterone synthesis in domestic ruminants. AB - Regulation of progesterone secretion from the corpus luteum during the oestrous cycle requires the integration of multiple signals to achieve the appropriate amount of progesterone to maximize reproductive efficiency. Development of a mature corpus luteum capable of secreting sufficient amounts of progesterone is dependent upon the pituitary hormones LH and growth hormone (GH). Continued secretion of progesterone from the mature corpus luteum is also dependent upon pituitary hormones. If pregnancy does not occur, prostaglandin F2 alpha (PGF2 alpha) of uterine origin causes a precipitous decrease in progesterone secretion and demise of the corpus luteum. A major point of regulation of progesterone secretion by both luteotrophic and luteolytic hormones appears to be regulation of transport of cholesterol through the mitochondrial membranes to cytochrome P450scc. It is likely that both luteotrophic and luteolytic hormones regulate steroidogenic acute regulatory protein (StAR), which facilitates transport. Regulation may be occurring through increases or decreases in gene transcription, translation efficiency or post-translational modifications such as phosphorylation. Thus, although synthesis of progesterone is a complex process, both positive and negative regulation of the process appears to occur primarily at a single step (transport of cholesterol to the inner mitochondrial membrane) in the pathway. PMID- 10692856 TI - Luteal peptides and their genes as important markers of ovarian differentiation. AB - Secreted peptide hormones and components of the steroidogenic machinery are molecules that are expressed usually in high amounts and in a time- and cell specific fashion within the cells that give rise to the bovine corpus luteum. They thus serve as useful markers for the events occurring within the nuclei of these cells that result in differentiation and the expression of the specific luteal phenotype. We have studied the bovine genes of three such luteal products: oxytocin, the new relaxin-like factor (RLF), and the steroidogenic acute regulatory protein (StAR). The oxytocin gene is expressed in the granulosal cells of the preovulatory follicle and in the large luteal cells of the immediately resulting early corpus luteum. The RLF gene is a major thecal cell product in antral and atretic follicles. It is also transcribed in luteal cells, but only in the mid- to late ovarian cycle and in pregnancy, following a temporal pattern of expression very similar to that of relaxin in pigs. The StAR gene appears to be upregulated only in the mid- to late ovarian cycle, several days after the increase in steroidogenic enzymes associated with luteinization and progesterone production. All three genes make use of the transcription factor SF-1 (Ad4BP) and, although they all respond to LH activation of adenylate cyclase, none utilize CRE-linked systems. Specific transcriptional activation must involve other factors to encode the information for the widely diverse temporal and cellular patterns of gene expression for these three genes. PMID- 10692857 TI - Intraovarian regulation of luteolysis. AB - The corpus luteum is a transient gland, which is only functional for 17-18 days in the cyclic cow or for up to 200 days in the pregnant cow. Regression of the corpus luteum is essential for normal cyclicity as it allows the development of a new ovulatory follicle, whereas prevention of luteolysis is necessary for the maintenance of pregnancy. Evidence acquired over the past three decades indicated that PGF2 alpha is the luteolytic hormone in ruminants. Nevertheless, the detailed mechanisms of PGF2 alpha action are just beginning to be clarified. A pivotal role for an endothelial cell product endothelin 1 (ET-1) has been documented in PGF2 alpha-induced luteal regression. ET-1 inhibited progesterone production by luteal cells in a dose-dependent manner via selective ET-1 binding sites (ETA). The inhibitory action of PGF2 alpha on progesterone secretion (in vivo and in vitro) was blocked by a selective ETA receptor antagonist. This implied that ET-1 (through ETA receptors present on steroidogenic cells) may have mediated the inhibitory effect of PGF2 alpha. The involvement of ET-1 in luteal regression was also suggested by the observation that the highest concentrations of ET-1 coincide with uterine PGF2 alpha surges. Furthermore, PGF2 alpha administration upregulated ET-1 expression within the corpus luteum. Later stages of luteal regression, which involve programmed cell death (PCD), are presumably mediated by immune cells. ET-1 may also be involved in this process by promoting leukocyte migration and stimulating macrophages to release tumour necrosis factor alpha (TNF alpha). The TNF alpha receptor type 1 (p55) is present on luteal cells (endothelial and steroidogenic cells) and could initiate PCD and the structural demise of the corpus luteum. PMID- 10692858 TI - Regulation of gonadotrophin-releasing hormone secretion by testosterone in male sheep. AB - In males, including the ram, testosterone, acting via its primary metabolites oestradiol and dihydrotestosterone (DHT), suppresses circulating LH concentrations. This effect is due primarily, although not totally, to decreased frequency of gonadotrophin-releasing hormone (GnRH) pulses. The arcuate ventromedial region (ARC-VMR) of the mediobasal hypothalamus and possibly the medial preoptic area (mPOA) are sites at which oestradiol acts to suppress GnRH, but the site of DHT action is not known. Given that native GnRH neurones appear to contain few or no oestrogen or androgen receptors, the effects of testosterone metabolites probably are exerted by modulating activity of inhibitory interneurone systems such as beta-endorphin, dopamine, and gamma-aminobutyric acid (GABA). Although beta-endorphin clearly inhibits GnRH secretion, the observation that testosterone treatment during a long-day photoperiod reduced proopiomelanocortin (POMC) mRNA in the arcuate nucleus while coincidentally suppressing GnRH release indicates that beta-endorphin does not mediate the inhibitory effect of testosterone on GnRH. Activation of GABAA receptors in either the mPOA or ARC-VMR suppressed LH, whereas activation of GABAB receptors in the ARC-VMR increased LH pulse amplitude. Therefore, it is suggested that GABA acts in both regions to regulate LH. Whereas testosterone affects GABA metabolism in the rat hypothalamus, its effect in the ram hypothalamus is yet to be determined. Testosterone treatment activated dopaminergic cells in the retrochiasmatic A15 area in the same animals in which it suppressed POMC mRNA in the arcuate nucleus. This dopaminergic system may partially mediate the negative feedback effect of testosterone in the ram analogous to its role in partially mediating the negative effect of oestrogen in the ewe. Future studies must concentrate on determining how these and other putative inhibitory neuronal systems interact and how they in turn are regulated by environmental factors such as photoperiod. PMID- 10692859 TI - Role of male-female interaction in regulating reproduction in sheep and goats. AB - The induction of synchronous ovulatory activity in anovulatory sheep and goats after the introduction of males, the 'male effect', has probably been used to advantage since these species were domesticated and the underlying physiological and behavioural mechanisms have been progressively elucidated over the past 50 years. Less well understood is the analogous effect of oestrous females on males. This review examines the nature and importance of these male-female interactions in sheep and goats, and describes the most important internal and external factors influencing the reproductive outcomes of such interactions. It is proposed that the male and female effects are both components of a self reinforcing cycle of stimulation that, under ideal conditions, culminates in the synchronous very rapid onset (within days) of fertile reproductive activity. However, precisely because of the speed of this response, it is suggested that mechanisms have evolved to limit its efficacy, and thus prevent conception at inappropriate times. The complexity of these factors and the interactions between them are highlighted, and a broad conceptual framework for understanding them is proposed based upon an appreciation of variation in both the responsiveness of the target animal and the quality of the signal from the signalling animal. PMID- 10692860 TI - Sexual behaviour of rams: male orientation and its endocrine correlates. AB - The components of heterosexual behaviour in rams are reviewed as a basis for understanding partner preference behaviour. A small percentage of rams will not mate with oestrous females and if given a choice will display courtship behaviour towards another ram in preference to a female. Some of the endocrine profiles of these male-oriented rams differ from those of heterosexual controls. These differences include reduced serum concentrations of testosterone, oestradiol and oestrone, reduced capacity to produce testosterone in vitro, and reduced capacity to aromatize androgens in the preoptic-anterior hypothalamus of the brain. Our observation that aromatase activity is significantly lower in the preoptic anterior hypothalamic area of male-oriented rams than in female-oriented rams may indicate an important neurochemical link to sexual behaviour that should be investigated. The defect in steroid hormone production by the adult testes of the male-oriented ram may represent a defect that can be traced to the fetal testes. If this contention is correct, partner preference behaviour of rams may also be traceable to fetal development and represent a phenomenon of sexual differentiation. PMID- 10692861 TI - The functional integrity and fate of cryopreserved ram spermatozoa in the female tract. AB - Cryopreservation advances capacitation-like changes in ram spermatozoa. These changes are reflected in an increased fertilizing ability compared with fresh spermatozoa, followed by an accelerated decline in fertilizing ability after incubation in vitro or in vivo. Furthermore, frozen-thawed spermatozoa are released earlier than fresh spermatozoa after binding to oviduct cells in vitro, confirming their physiological readiness to participate in fertilization despite their short lifespan. After insemination large numbers of spermatozoa are lost from the female reproductive tract of the ewe via the vagina. Frozen-thawed spermatozoa are expelled faster than fresh spermatozoa. The advanced membrane status of frozen-thawed spermatozoa may provoke their rapid loss and possibly makes them more vulnerable to attack by uterine leucocytes, or by some other mechanism, as a high proportion of spermatozoa lost from the tract are decapitated. The observed destabilization of the membranes of cryopreserved spermatozoa is accompanied by impaired sperm transport, associated with mitochondrial injury, necessitating intrauterine deposition of frozen-thawed semen to obtain satisfactory fertility after artificial insemination. However, the frozen-thawed spermatozoa that can participate in fertilization may contribute to increased embryonic loss by the advancement of cleavage or through a direct effect of cryopreservation on the male genome. PMID- 10692862 TI - Uterine differentiation as a foundation for subsequent fertility. AB - Uterine differentiation in cattle and sheep begins prenatally, but is completed postnatally. Mechanisms regulating this process are not well defined. However, studies of urogenital tract development in murine systems, particularly those involving tissue recombination and targeted gene mutation, indicate that the ideal uterine organizational programme evolves epigenetically through dynamic cell-cell and cell-matrix interactions that define the microenvironmental context within which gene expression occurs and may ensure adult tissue stability. In the cow and ewe, transient postnatal exposure of the developing uterus to steroids can produce immutable changes in adult uterine tissues that may alter the embryotrophic potential of the uterine environment. Thus, success of steroid sensitive postnatal events supporting uterine growth and development can dictate the functional potential of the adult uterus. Studies to determine effects of specific steroidal agents on patterns of uterine development during defined neonatal periods, as well as the functional consequences of targeted neonatal steroid exposure in the adult uterus, should enable identification of critical developmental mechanisms and determinants of uterine integrity and function. Extreme adult uterine phenotypes (lesion models) created in cattle and sheep by strategic postnatal steroid exposure hold promise as powerful tools for the study of factors affecting uterine function and the rapid identification of novel uterine genes. PMID- 10692863 TI - IGF paracrine and autocrine interactions between conceptus and oviduct. AB - Development in vitro is influenced by embryo density, serum, somatic cell co culture and the production of 'embryotrophic' paracrine and autocrine factors. Research in our laboratory has focussed principally on the insulin-like growth factor (IGF) family. We have demonstrated that pre-attachment bovine and ovine embryos express mRNAs encoding a number of growth factor ligand and receptor genes including all members of the IGF ligand and receptor family throughout this developmental interval. In addition, early embryos express mRNAs encoding IGF binding proteins (IGFBPs) 2-5 from the one-cell to the blastocyst stage and IGFBP5 mRNA at the blastocyst stage. Cultured bovine blastocysts release up to 35 pg per embryo in 24 h, whereas release of IGF-I was below detectable values. Analysis extended to bovine oviductal cultures has also demonstrated that mRNAs encoding these IGF family members are present throughout an 8 day culture period. Transcripts encoding IGFBPs 2-6 were also present. Release of both IGFs was recorded over an 8 day culture period. IGF-II release was significantly greater than that observed for IGF-I. Therefore, the IGFs are present throughout the maternal environment during early embryo development. The oocyte, within the follicle, is held in an environment high in IGFs and IGFBPs. The zygote, after fertilization, is maintained in an IGF-rich environment while free-living in the oviduct and the uterus. This review is focused on the IGF family and IGFBPs and their roles in enhancing development up to the blastocyst stage. PMID- 10692864 TI - The regulation of interferon-tau production and uterine hormone receptors during early pregnancy. AB - During early pregnancy the bovine embryo must produce a protein called interferon tau which inhibits the development of the luteolytic mechanism. Failure to inhibit luteolysis is the major cause of pregnancy loss in cows. The embryo must produce sufficient quantities of interferon tau by about day 16 to prevent luteolysis. Its ability to achieve this is largely dependent on the pattern of maternal progesterone production. A late rise in progesterone after ovulation or poor progesterone secretion during the luteal phase results in the development of poor embryos capable of producing little or no interferon tau at the critical time. The embryo inhibits luteolysis by preventing development of oxytocin receptors on the luminal epithelium of the uterine endometrium and thus oxytocin induced secretion of PGF2 alpha and by the induction of a prostaglandin synthesis inhibitor within the endometrium. In sheep it has been hypothesised that interferon tau acts to inhibit endometrial oestrogen receptors and thus oestrogen induced up-regulation of oxytocin receptors. In cows, the embryo inhibits the development of oxytocin receptors and the initiation of luteolysis without causing any change in uterine oestrogen receptors. Thus in the cow, the mechanism by which interferon tau inhibits oxytocin receptor development remains to be determined. PMID- 10692866 TI - Roles of extracellular matrix in follicular development. AB - The cellular biology and changes in the extracellular matrix of ovarian follicles during their development are reviewed. During growth of the bovine ovarian follicle the follicular basal lamina doubles 19 times in surface area. It changes in composition, having collagen IV alpha 1-26 and laminin alpha 1, beta 2 and gamma 1 at the primordial stage, and collagen IV alpha 1 and alpha 2, reduced amounts of alpha 3-alpha 5, and a higher content of laminin alpha 1, beta 2 and gamma 1 at the antral stage. In atretic antral follicles laminin alpha 2 was also detected. The follicular epithelium also changes from one layer to many layers during follicular growth. It is clear that not all granulosal cells have equal potential to divide, and we have evidence that the granulosal cells arise from a population of stem cells. This finding has important ramifications and supports the concept that different follicular growth factors can act on different subsets of granulosal cells. In antral follicles, the replication of cells occurs in the middle layers of the membrana granulosa, with older granulosal cells towards the antrum and towards the basal lamina. The basal cells in the membrana granulosa have also been observed to vary in shape between follicies. In smaller antral follicles, they were either columnar or rounded, and in follicles > 5 mm the cells were all rounded. The reasons for these changes in matrix and cell shapes are discussed in relation to follicular development. PMID- 10692865 TI - Mechanism of action of interferon-tau in the uterus during early pregnancy. AB - Early pregnancy is maintained in ruminants through the actions of conceptus derived interferon (IFN)-tau on the endometrium. IFN-tau alters uterine release of PGF2 alpha' which results in rescue of the corpus luteum and continued release of progesterone. The mechanism of action of IFN-tau includes inhibition of oestradiol receptors, consequent reduction in oxytocin receptors, activation of a cyclooxygenase inhibitor, and a shift in the PGs to favour PGE2 over PGF2 alpha' IFN-tau also induces several endometrial proteins that may be critical for survival of the developing embryo. One endometrial protein induced by pregnancy and IFN-tau has been identified as bovine granulocyte chemotactic protein-2 (bGCP 2). This chemotactic cytokine (chemokine) has been used as a marker to delineate IFN-tau from IFN-alpha responses in the endometrium. A second protein, called ubiquitin cross-reactive protein (UCRP), resembles a tandem ubiquitin repeat. UCRP becomes conjugated to cytosolic endometrial proteins in response to IFN-tau and pregnancy. Proteins conjugated to UCRP are either modulated or targeted for processing through the proteasome. The action of IFN-tau is mediated by induction of signal transducer and activator of transcription 1 (STAT-1), STAT-2 and interferon regulatory factor 1 (IRF-1) transcription factors. Induction of these transcription factors, the alpha chemokines and UCRP is the prelude to maternal recognition of pregnancy in ruminants. PMID- 10692867 TI - Plasmin-tumour necrosis factor interaction in the ovulatory process. AB - Collagen breakdown and apoptotic cell death within the apex of the preovulatory ovine follicle are hallmarks of impending ovarian rupture. An integrative mechanism is proposed whereby gonadotrophic stimulation of urokinase-type plasminogen activator secretion by the follicular-contiguous ovarian surface epithelium elicits a localized increase in tissue plasmin, which activates collagenolysis and tumour necrosis factor alpha-induced cell death within the formative ovulatory stigma. PMID- 10692868 TI - Growth factors and extracellular matrix proteins in interactions of cumulus oocyte complex, spermatozoa and oviduct. AB - The expression and localization of selected growth factor systems and extracellular matrix (ECM) components that may influence oocyte maturation and fertilization within the mammalian oviduct are reported. Fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) systems could be detected by use of RT-PCR, RNase protection assay (RPA) and immunohistochemistry in bovine follicles, bovine cumulus-oocyte complexes (COC) and bovine and marmoset oviducts. Two different subtypes of the FGF receptor (FGFR-1 and -2) were identified in distinct cell types, indicating a functional difference. A complete epidermal growth factor (EGF) system was found in the porcine, but not in the bovine, oviduct. There were additional differences between bovine and primate oviducts: FGF-1/2 and FGFR were increased in the marmoset around ovulation, in contrast to an increase in FGF-1 in the cow. Immunohistochemistry revealed accumulation and storage of FGF and VEGF on the surface of the epithelium, possibly due to their binding property on heparanglycoproteins. Other ECM components, matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase 1 (TIMP-1), were found to be modulated in the ovarian follicle, COC and oviduct during the cycle. An oviduct-mediated depletion of sperm surface proteins (BSP1-3) was discovered as well as a sperm-induced novel oviductal mRNA related to an anti-oxidant protein family. Associated systems of growth factors and ECM components can be suggested as paracrine or autocrine mediators during fertilization in a species-, cycle- and tissue-dependent manner. PMID- 10692869 TI - Regulation of ovarian extracellular matrix remodelling by metalloproteinases and their tissue inhibitors: effects on follicular development, ovulation and luteal function. AB - In most organs, remodelling of tissues after morphogenesis is minimal; however, normal ovarian function depends upon cyclical remodelling of the extracellular matrix (ECM). The ECM has a profound effect on cellular functions and probably plays an important role in the processes of follicular development and atresia, ovulation, and development, maintenance and regression of corpora lutea. Matrix metalloproteinases (MMPs; collagenases, gelatinases, stromelysins and membrane type MMPs) cleave specific components of the ECM and are inhibited by tissue inhibitors of metalloproteinases (TIMPs). MMPs have been detected at all stages of follicular development and probably modulate follicular expansion or atresia within the ovarian stroma. In addition, increased MMP activity appears to be required for ovulation since follicular rupture occurred in the absence of plasminogen activator activity and inhibitors of MMPs blocked follicular rupture. Development and luteolysis of the corpus luteum are accompanied by extensive remodelling of the ECM. Differentiation and regression of luteal cells are associated with construction and degradation of ECM, respectively. There is increasing evidence that ECM components enhance luteinization; whereas loss of ECM results in luteal cell death. Ovine large luteal cells may be the primary type of cell responsible for controlling the extent of remodelling of luteal ECM since they produce TIMP-1, TIMP-2 and plasminogen activator inhibitor 1. The ratio of active MMPs to TIMPs may be important in maintaining an ECM microenvironment conducive to the differentiation of follicular-derived cells into luteal cells, and maintenance of the phenotype of luteal cells. PMID- 10692870 TI - Nutrition and fetal growth: paradoxical effects in the overnourished adolescent sheep. AB - Inappropriate maternal nutrient intake at key developmental timepoints during ovine pregnancy has a profound influence on the outcome of pregnancy and aspects of postnatal productivity. However, the responses to alterations in maternal nutrition in adult sheep are often highly variable and inconsistent between studies. The growing adolescent sheep provides a new, robust and nutritionally sensitive paradigm with which to study the causes, consequences and reversibility of prenatal growth restriction. Overnourishing the adolescent dam to promote rapid maternal growth throughout pregnancy results in a major restriction in placental mass, and leads to a significant decrease in birthweight relative to moderately fed, normally growing adolescents of equivalent gynaecological age. Maternal insulin and IGF-I concentrations are increased from an early stage of gestation in overnourished adolescent dams and these hormones ensure that the anabolic drive required to promote maternal tissue synthesis is initiated at a time when the nutrient requirements of the gravid uterus are low. The major restriction in fetal growth in rapidly growing dams occurs irrespective of high concentrations of essential nutrients in the maternal circulation and suggests that the small size or altered metabolic and transport capacity of the placenta is the primary constraint to fetal growth. The decrease in placental weight in the overnourished animals reflects a significant reduction in both fetal cotyledon number and mean cotyledon weight. The role of nutritionally mediated alterations in progesterone and the components of the IGF system in this early pregnancy placental phenomenon are being investigated. Nutritional switch-over studies have demonstrated that reducing maternal nutrient intake at the end of the first third of pregnancy can stimulate placental growth and enhance pregnancy outcome, but increasing nutrient intake at this time has a deleterious effect on placental development and fetal growth. PMID- 10692871 TI - Placental transport of nutrients and its implications for fetal growth. AB - Placental growth during early and mid-pregnancy has a powerful, constraining influence on fetal growth during late pregnancy. Studies involving surgical and environmental reduction of placental size in sheep have shown an associated reduction in capacity to transport oxygen, glucose and amino acids. Oxygen transport is limited by placental blood flow but transport of glucose and amino acids is determined by the abundance and activity of specific transport proteins. Glucose transporters include the GLUT1 and GLUT3 isoforms previously identified in brain and other tissues; systems for active transport of amino acids have been inferred but not characterized. Placental metabolism of glucose and amino acids has major effects both on the quantity of carbon and nitrogen delivered to the fetus, and on the composition of substrates involved. For example, the uteroplacental tissues consume more than 60% of uterine glucose uptake during late pregnancy, and the placenta substantially modifies the pattern of amino acids delivered to fetal blood. The placenta also participates in the array of metabolic adaptations of maternal and conceptus tissues to altered maternal nutrient supply. Placental capacity for glucose transport in moderately undernourished ewes is upregulated, partly by increased expression of the GLUT3 transport protein. During more severe glucose deprivation, placental transfer and fetal uptake of glucose are constrained in proportion with maternal supply, leading to fetal growth retardation. PMID- 10692872 TI - Effects of energy balance on follicular development and first ovulation in postpartum dairy cows. AB - As milk production has increased during the past four decades, conception rates in lactating cows have declined. Although reduced reproductive performance has been associated with high milk yields, measures of postpartum ovarian activity have been more closely related to energy balance. The relationship between daily energy balance and postpartum reproductive activity is confirmed by longer intervals to first ovulation in cows with greater body condition loss. Patterns in daily energy balance, such as improvement from nadir, have been correlated with enhanced follicular function and a shorter interval to first ovulation. Such observations are consistent with increased LH pulse frequency following the energy balance nadir in lactating dairy cows. Evidence indicates a primarily hypothalamic locus for the modulation of LH secretion during negative energy balance. Formation of follicular waves after parturition begins synchronously in response to increased FSH in the first week postpartum, and is typically not a limiting factor in reproductive recrudescence. Altered follicular responsiveness to gonadotrophic support through changes in metabolic hormones such as insulin like growth factor I (IGF-I) and insulin may contribute to impaired function of dominant follicles early postpartum. Positive relationships between changes in energy balance, peripheral IGF-I and function of dominant follicles support the identification of IGF-I and the day of the energy balance nadir as metabolic modulators of postpartum ovarian activity in dairy cows. PMID- 10692873 TI - The role of leptin in nutritional status and reproductive function. AB - Infertility associated with suboptimal nutrition is a major concern among livestock producers. Undernourished prepubertal animals will not enter puberty until they are well fed; similarly, adult, normally cyclic females will stop cycling when faced with extreme undernutrition. Work in our laboratory has focused on how body fat (or adiposity) of an animal can communicate to the brain and regulate reproductive competence. In 1994, the discovery in rodents of the obese (ob) gene product leptin, secreted as a hormone from adipocytes, provided a unique opportunity to understand and hence regulate whole body compositional changes. There is now evidence that similar mechanisms are functioning in livestock species in which food intake, body composition, and reproductive performance are of considerable economic importance. Leptin has been reported to be a potent regulator of food intake and reproduction in rodents. There is evidence indicating that at least some of the effects of leptin occur through receptor-mediated regulation of the hypothalamic protein neuropeptide Y (NPY). NPY is a potent stimulator of food intake, is present at high concentrations in feed-restricted cattle and ewes, and is an inhibitor of LH secretion in these livestock species. In our investigations in sheep, we have cloned a partial cDNA corresponding to the ovine long-form leptin receptor, presumably the only fully active form, and have localized the long-form leptin receptor in the ventromedial and arcuate nuclei of the hypothalamus. Leptin receptor mRNA expression was colocalized with NPY mRNA-containing cell bodies in those regions. We have also determined that hypothalamic leptin receptor expression is greater in feed restricted ewes than in well-fed ewes. These observations provide a foundation for future investigations into the nutritional modulators of reproduction in livestock. PMID- 10692874 TI - Activation of primordial follicles in vitro. AB - The resting pool of primordial follicles in mammalian ovaries is a potential resource for the genetic manipulation of domestic animals, the preservation of endangered species, and the amelioration of some forms of infertility in humans. Exploitation of this large reservoir of follicles depends on the development of methods for activating primordial follicles to begin growth in vitro and of methods for sustaining follicular growth to the stage at which oocytes are capable of meiotic maturation, fertilization and development to live young. It has been shown that primordial follicles of rodents, cattle and primates can initiate growth in vitro, even in serum-free medium. The signals that cause primordial follicles to leave the resting pool or remain quiescent are unknown. However, of interest is the observation that in cultures of whole rodent ovaries an apparently normal number of follicles leaves the resting pool and begins to grow, whereas in cultures of isolated bovine or primate ovarian cortex almost all primordial follicles activate and develop into primary follicles. This finding suggests that non-cortical portions of the ovary may regulate the flow of follicles from the resting reservoir. In cattle, it has been difficult to sustain follicular growth beyond the primary stage and the development of methods for doing so are critical for achievement of the practical goal of use of the primordial pool for embryo production. However, the development of murine follicles in vitro from the primordial stage through oocyte maturation and fertilization, and the birth of one pup, provides encouragement for efforts to achieve similar results in large mammals. PMID- 10692875 TI - Aspects of follicular and oocyte maturation that affect the developmental potential of embryos. AB - The ability to mature, be fertilized and finally to develop into a viable embryo is acquired gradually by the oocyte during progressive differentiation throughout folliculogenesis. This process starts with oocyte growth during the first steps of follicular development. As the oocyte is close to its final size, other modifications occur, less spectacular but at least as important in determining the resulting ability of the oocyte to accomplish its reproductive purpose (developmental competence). These modifications, referred to as 'oocyte capacitation', are probably influenced by the follicle. The proportion of developmentally competent oocytes increases with follicular size. However, the relationship between follicular growth and oocyte competence is not very strict, since a given oocyte may acquire its competence at any stage of follicular growth and since some examples of functional disjunction between follicular size and oocyte competence are described. Follicular atresia may impair the acquisition of oocyte competence, as evidenced by the parallel study of follicular characteristics and of the developmental potential of their oocytes treated individually through in vitro maturation, fertilization and development. However, when atresia is experimentally induced in large preovulatory follicles, oocytes remain competent, indicating that once competence is acquired, it is no longer sensitive to atresia. Oocyte maturation represents only the end of this long and progressive process and validates the preparation of the oocyte by conferring its final developmental ability. As evidenced by recent cloning experiments, the cytoplasmic aspects of oocyte maturation are crucial for the acquisition of developmental competence. This cytoplasmic maturation may be activated in vitro by the use of complex media supplement (serum, follicular fluid) but the use of defined media for maturation allowed the identification of some active factors (such as epidermal growth factor, growth hormone, inhibin and activin). The study of some differential models of oocyte competence (follicular size and atresia, Booroola gene, prepubertal oocytes) will provide a better understanding of oocyte capacitation and maturation, and allow the improvement of in vitro methods for oocyte maturation, which represent the most limiting step of in vitro production of embryos in large mammals. PMID- 10692876 TI - Development of serum-free culture systems for the ruminant embryo and subsequent assessment of embryo viability. AB - The mammalian embryo undergoes considerable changes in its physiology and energy metabolism as it proceeds from the zygote to the blastocyst stage. Complete development of the mammalian zygote in vitro was restricted to a few strains of mice and their F1 hybrids for many years, as the ruminant embryo arrested development at the 8- to 16-cell stage. The introduction of co-culture of ruminant embryos with somatic cells in the mid-1980s helped to alleviate this in vitro induced arrest. However, such culture systems required the use of complex tissue culture media and serum. Serum has subsequently been shown to induce several abnormalities during embryo development in culture and has been associated with the production of offspring with significantly greater birth weights than normal, leading to both difficulties in pregnancy management and an unacceptable frequency of neonatal death. Resurgence of interest in mammalian embryo physiology has culminated in the formulation of defined embryo culture media, capable of supporting a high percentage of viable blastocyst development in vitro. Optimum embryo development in culture has been shown to take place not in one, but two or more media, each designed to cater for the changing requirements and metabolism of the embryo as it develops. The development of viability assays to identify those embryos with the highest developmental potential will further increase the efficiency of embryo transfer procedures. Assays based upon nutrient uptake and subsequent utilization make promising candidates. PMID- 10692877 TI - Sexing mammalian spermatozoa and embryos--state of the art. AB - Methods for sexing preimplantation embryos range from karyotyping to recording speed of development in vitro. The only method used routinely on a commercial scale is to biopsy embryos and amplify Y-chromosome-specific DNA using the polymerase chain reaction. This method is effective for more than 90% of embryos and is > 95% accurate. Within males, spermatozoa are essentially identical phenotypically due to: (1) connection of spermatogenic cells by intercellular bridges, (2) transcriptional inactivation of sex chromosomes during meiosis and spermiogenesis, (3) severe limitation of all gene expression during the later stages of spermiogenesis, and (4) coating all spermatozoa with common macromolecules during and after spermiogenesis. One consequence is that no convincing phenotypic difference has been detected between X- and Y-chromosome bearing spermatozoa. The only consistently successful, nondestructive approach to sexing spermatozoa is to quantify DNA in spermatozoa using a fluorescing DNA binding dye followed by flow cytometry and cell sorting. X-chromosome-bearing ruminant spermatozoa have about 4% more DNA compared with Y-chromosome-bearing spermatozoa; accuracy of sorting can exceed 90% routinely, and sorting rates currently exceed 10(3) live spermatozoa of each sex chromosome composition s-1. Hundreds of apparently normal offspring from a number of species have been produced from sexed semen, some via intrauterine artificial insemination. PMID- 10692878 TI - Nuclear transfer from somatic cells: applications in farm animal species. AB - The reconstruction of mammalian embryos by transfer of a blastomere nucleus to an enucleated oocyte or zygote allows for the production of genetically identical individuals. This has advantages for research (that is, as biological controls) and commercial applications (that is, multiplication of genetically valuable livestock). However, the number of offspring that can be produced from a single embryo is limited both by the number of blastomeres (embryos at the 32-64-cell stage are the most widely used in farm animal species) and the limited efficiency of the nuclear transfer procedure. The ability to produce live offspring by nuclear transfer from cells that can be propagated and maintained in culture offers many advantages, including the production of many identical offspring over an extended period (since cultured cells can be frozen and stored indefinitely) and the ability to modify genetically or to select populations of cells of specific genotypes or phenotypes before embryo reconstruction. This objective has been achieved with the production of lambs using nuclei from cultured cells established from embryonic, fetal and adult material. In addition, lambs transgenic for human factor IX have been produced from fetal fibroblasts transfected and selected in culture. PMID- 10692879 TI - Death in custody. PMID- 10692880 TI - Nutritional causes of impaired fetal growth and their treatment. PMID- 10692881 TI - Multiple chemical sensitivity--is the environment really to blame? PMID- 10692882 TI - Imprecision in medical communication: study of a doctor talking to patients with serious illness. AB - Uncertainty is believed to be a central feature in illness experiences. Conversations between a consultant hematologist and 61 seriously ill patients were transcribed, entered on a database and scrutinized for patterns of language uncertainty by linguistic concordancing analysis. Transcripts were then discussed in detail with the hematologist, and techniques of protocol analysis were used to gain insight into his thought processes during consultations. The main findings were that the doctor used many more expressions of uncertainty than did patients: that evaluative terms were widely used to reassure rather than to worry patients; and that patients and doctor together used certain key terms ambiguously, in a manner which allowed the doctor to feel that facts were not misrepresented while perhaps permitting the patient to feel reassured. PMID- 10692883 TI - Attitudes to organ donation among South Asians in an English high street. AB - In the UK, people of South Asian origin are at more than twice the risk of end stage renal failure encountered in the Caucasian population but are under represented among organ donors. Difficulties with matching mean that few donated kidneys are suitable for transplantation to South Asian recipients. A survey of attitudes in 100 South Asian adults was conducted in the main street of Southall, Middlesex. 90 of those questioned were aware of organ transplantation and 69 had heard about donor cards. However, the 16% who carried a donor card was lower than the 28% reported in the general population. The main reason for the low organ donation rate by South Asians seemed to be lack of knowledge, and this could be remedied by more targeting of information in the Asian media. PMID- 10692884 TI - Hysterosalpingo contrast sonography as a screening test for tubal patency in infertile women. AB - The most informative method for assessing tubal patency in subfertile women is laparoscopy-and-dye. This investigation, however, puts a large burden on services and a screening test is needed that identifies a high likelihood of occlusion. In our infertility programme we introduced hysterosalpingo contrast sonography for this purpose, operated entirely by ultrasonographers. A series of audits indicated that this innovation speeded the process of investigation by several weeks and reduced the number of laparoscopy-and-dye procedures by 75%. The negative predictive value was 89% and the positive predictive value was 44%. The main limitation of the method was the long period required for training, in those without extensive experience of vaginal ultrasonography. PMID- 10692885 TI - The dos and don'ts of examining the respiratory system: a survey of British Thoracic Society members. PMID- 10692886 TI - Recurrent renal cholesterol embolism. PMID- 10692887 TI - Systemic vasculitis with multiple aneurysms complicating systemic lupus erythematosus. PMID- 10692888 TI - Aortic dissection with chronic haemopericardium. PMID- 10692889 TI - Appendico-cutaneous fistula. PMID- 10692890 TI - Caduceus, porcelain and palette: John Wall of Worcester. PMID- 10692891 TI - I drink, therefore I am: alcohol and creativity. PMID- 10692892 TI - Neurology in the National Gallery. PMID- 10692893 TI - Multicentre research ethics committees. PMID- 10692894 TI - Severe parkinsonism secondary to carbon monoxide poisoning. PMID- 10692895 TI - Antioxidants in wine and tea. PMID- 10692896 TI - Transport and temperature effects on blood potassium. PMID- 10692897 TI - Transport and temperature effects on blood potassium PMID- 10692898 TI - Infections prevent the development of asthma--true, false or both? PMID- 10692899 TI - Multicentre research ethics committees: have they helped? PMID- 10692900 TI - Replacement of damaged neural cells: a mirage? PMID- 10692901 TI - Renal artery stenosis as a cause of renal impairment: implications for treatment of hypertension and congestive heart failure. PMID- 10692902 TI - Placebos and placebo effects in medicine: historical overview. PMID- 10692903 TI - Management of acute bursitis: outcome study of a structured approach. AB - In patients with septic bursitis the indications for admission and surgical intervention remain unclear, and practice has varied widely. The effectiveness of a conservative outpatient based approach was assessed by an outcome study in a prospective case series. Consecutive patients attending an emergency department with acute swelling of the olecranon or prepatellar bursa were managed according to a structured approach, subjective and objective outcomes being assessed after two to three days, and subsequently as required until clinical discharge. Long term outcomes were assessed by telephone follow-up for up to eighteen months. 47 patients were included in the study: 22 had septic bursitis, 15 of the olecranon bursa and 7 of the prepatellar bursa. The mean visual analogue pain scores of those with septic bursitis improved from 4.8 at presentation to 1.7 at first follow-up for olecranon bursitis, and from 3.8 to 2.7 for prepatellar bursitis. Symptoms improved more slowly for patients with non-septic bursitis. No patients were admitted initially, but 2 were admitted (two days each) after the first follow-up appointment. One patient had incision and drainage on the third attendance, and 3 patients developed discharging sinuses, which all healed spontaneously. All patients made a good long-term symptomatic recovery and all could lean on the elbow or kneel by the end of the follow-up period. The management protocol, with specific criteria for admission and surgical intervention, thus produced good results with little need for operation or admission. PMID- 10692904 TI - Switching to statins: a challenge for primary care. AB - In 1997, doctors in England received official guidelines on the use of statins (3 hydroxy-3-methylglutaryl coenzyme A inhibitors) for primary and secondary prevention of coronary heart disease (CHD). Six months later we determined the status of patients who had been discharged from a specialist lipid clinic in 1989. 195 patients received questionnaires, with the consent of their general practitioners, regarding morbidity in, the subsequent decade and present medication, and were asked to have their cholesterol checked. Analysis was confined to the 86 with a current cholesterol measurement. Of 61 patients who had been discharged on a regimen of dietary advice and/or medication for primary prevention of CHD, 8 had been changed to a statin and 6 had been started on one. According to the new guidelines, none of these qualified for treatment. Of 25 patients who had been discharged on drugs for secondary prevention, all qualified for a statin but only 14 were receiving one--in 6 cases without achieving the recommended reductions in cholesterol. In many of the patients reviewed, treatment had not been altered to conform with the new guidelines. If hyperlipidaemic patients are to benefit promptly from advances in treatment, one solution might be a central registry that arranged regular tests and reported back to general practitioners. However, since many patients at risk do not have very high cholesterol levels, a coordinated approach to CHD risk factors would be preferable. PMID- 10692905 TI - Family support in general practice. AB - At a time when social services are overburdened in Britain, family support in general practice offers one way to fill the gap. In the Well Family Project, a 'family support coordinator' worked within a general practice in Hackney, London. In the first eighteen months she saw 113 clients. Evaluation was by semistructured interviews with a sample of these clients and with professional workers. Comments from those interviewed indicate that the family support was valued. The general practice base was convenient and non-stigmatizing. By adopting a proactive approach, the project was able to work with clients who had previously 'slipped through the net'. Some of the professionals interviewed would have liked to provide the same help, but were unable to do so because of time and other constraints. Family support provided through general practice was well received by vulnerable families. Although there was overlap with the remit of health visitors and social workers, the protected time and the independence of the coordinator enabled clients to obtain the help they wanted. The replicability of this strategy now needs to be assessed. PMID- 10692906 TI - Severe parkinsonism secondary to carbon monoxide poisoning. PMID- 10692907 TI - Excision of a false left ventricular aneurysm. PMID- 10692908 TI - Heart valve disease in seronegative rheumatoid arthritis. PMID- 10692909 TI - Splenic perivascular fibrosis as an ultrasound finding. PMID- 10692910 TI - The English inoculator: Jan Ingen-Housz. PMID- 10692911 TI - Some unanswered questions about NICE. PMID- 10692912 TI - Tiger bites. PMID- 10692913 TI - Antenatal screening for HIV. PMID- 10692914 TI - Coeliac disease in adults. PMID- 10692915 TI - Euthanasia in the Netherlands. PMID- 10692916 TI - Adverse reactions to herbal treatment. PMID- 10692917 TI - Autoimmune enteropathy with goblet cell antibodies. PMID- 10692918 TI - Contralateral extradural haematoma after ventriculoperitoneal shunt insertion. PMID- 10692919 TI - A Harley Street address PMID- 10692920 TI - Management of penile fracture. PMID- 10692921 TI - Glaucoma surgery in the United Kingdom: why, who and when. PMID- 10692922 TI - Can altering the pattern of laser photocoagulation for proliferative diabetic retinopathy help retain visual fields for driving? PMID- 10692923 TI - Bowman Lecture 1998. Diabetic retinopathy: some cellular, molecular and therapeutic considerations. PMID- 10692924 TI - The National Survey of Trabeculectomy. I. Sample and methods. AB - PURPOSE: The National Survey of Trabeculectomy was designed to evaluate current practices of glaucoma surgery in the United Kingdom and to determine the success and complication rates of trabeculectomy on a national basis. This paper reports the survey methods, levels of consultant activity, waiting times, indications for surgery and the demographic and clinical characteristics of the patient sample. METHODS: Consultant ophthalmologists performing trabeculectomy in the United Kingdom were studied. Four consecutive patients undergoing trabeculectomy under each consultant prior to 18 June 1996 were retrospectively sampled. Patients were followed prospectively and evaluated 6 and 12 months after surgery. Data were collected by self-administered postal questionnaires. To determine the effects of selection and reporting bias a validation study of 14 randomly selected units was also conducted. RESULTS: Three hundred and eighty-two consultants recruited 1454 eligible patients for analysis. The mean age of patients was 69.2 years (standard deviation 10.9) and 51.7% were male. The underlying diagnosis was primary open angle glaucoma in 89.2%, pseudoexfoliation glaucoma in 5.4%, normal tension glaucoma in 3.8% and pigmentary glaucoma in 1.6%. There was advanced visual field damage in 50.5% of the cohort by the time of listing. The main indications for surgery were failure of medication to control intraocular pressure in 57.1%, progressive visual field loss in 26.5% and progressive optic disc damage in 4.8%. Primary surgery was undertaken in 4.8% of patients. In 80% trabeculectomy was performed within 3 months of listing. However, almost a third of consultants considered individual patient's waiting time too long. Validation studies confirmed that systematic bias did not operate in the selection of patients for the survey or in the reporting of outcomes. CONCLUSION: The findings of this survey are representative of current practices of trabeculectomy by consultants throughout the United Kingdom and show considerable variation in practice. Failure to control intraocular pressure with topical medications was the main indication for surgery. Advanced glaucomatous visual field damage was present at the time of surgery in half the sample. Though most patients were operated on within 3 months of listing, almost a third of consultants considered the wait unacceptably long. PMID- 10692925 TI - Altering the pattern of panretinal photocoagulation: could the visual field for driving be preserved? AB - PURPOSE: To identify the area of retina required to provide the visual field for driving and to investigate whether the pattern of panretinal photocoagulation (PRP) for proliferative diabetic retinopathy could be altered to avoid treatment in this area whilst leaving the total number of burns constant. METHODS: A mathematical model of the emmetropic eye is used to calculate retinal dimensions corresponding to different angles of visual field. These are used to define retinal regions that correspond to the UK DVLC visual field criteria and regions that lie outside this area. Further calculation estimates the number of laser burns applied within these regions for both 500 microns and 200 microns diameter spot sizes and various burn spacings. RESULTS: Modelling of the number of burns applied in the normal pattern of PRP agrees with the number required to control proliferative retinopathy. Reducing burn spacing or extending treatment up to the ora serrata allows application of sufficient burns to control the disease without encroaching on areas of the retina that provide the driving field. CONCLUSION: It is theoretically possible to alter the pattern of PRP to avoid treatment in retinal areas concerned with the driving visual field whilst leaving the total number of burns constant. This suggests that a clinical trial of such a pattern PRP could be performed to assess adequate control of proliferative retinopathy along with preservation of the visual field required for driving. PMID- 10692926 TI - Low-dose midazolam infusion for oculoplastic surgery under local anesthesia. AB - PURPOSE: Oculoplastic surgery with infiltration of local anaesthesia at the operative site performed as a day-case procedure is both efficient and cost effective. Patients considered unsuitable for this because of fear or apprehension may, however, benefit from per-operative conscious sedation. We sought to study the efficacy and safety of this using midazolam, a water-soluble benzodiazepine. METHOD: We have performed a controlled clinical trial comparing the effect of a low-dose intravenous infusion of midazolam (0.2 mg/ml of normal saline at a rate of 1 mg/h) with saline placebo on 48 subjects undergoing oculoplastic surgery with local anaesthesia. Patients were given pre- and post operative questionnaires assessing, amongst other factors, anxiety levels, pain, degree of reported amnesia and psychomotor recovery. RESULTS: Using the low-dose midazolam infusion no adverse cardiorespiratory reactions occurred. Patients receiving midazolam reported remembering significantly less about their operation than controls (p = 0.04) and showed significantly lower state-anxiety after their operation than before (p < 0.02). This change was not noted in the placebo group. There was no significant difference in the psychomotor performance of patients given midazolam compared with controls 2 h after surgery. CONCLUSIONS: A low-dose continuous infusion of midazolam can be used to safely provide effective anxiolysis and conscious sedation with good psychomotor recovery during oculoplastic procedures in a day-case setting. PMID- 10692927 TI - Missed orbital wall blow-out fracture as a cause of post-enucleation socket syndrome. AB - BACKGROUND: Post-enucleation socket syndrome (PESS: deep upper lid sulcus, ptosis or upper lid retraction, enophthalmos and lower lid laxity) is a well-recognised complication of a volume-deficient anophthalmic socket. A patient requiring enucleation following severe ocular trauma may have an underlying orbital wall blow-out fracture which if overlooked can cause severe volume deficit with poor cosmesis and limited prosthesis motility. PURPOSE: To establish the prevalence of an undiagnosed blow-out fracture in patients with PESS and a history of relevant trauma. METHODS: Medical records and orbital computed tomography (CT) scans were reviewed for all patients presenting with PESS and a history of relevant trauma. RESULTS: Undiagnosed blow-out fractures were found in 15 (33%) of 45 patients presenting between August 1993 and December 1996. These were significant enough to warrant surgical repair in 13 (29%) patients. CONCLUSIONS: We suggest that any patient presenting with PESS and a history of relevant trauma should be considered to have an orbital wall blow-out fracture until proven otherwise by CT scanning of the orbit. Similarly any patient requiring enucleation following severe ocular trauma should undergo CT scanning to rule out a coexisting blow-out fracture which could be repaired at the time of enucleation. PMID- 10692928 TI - Corneal biopsy with tissue micro-homogenisation for isolation of organisms in bacterial keratitis. AB - PURPOSE: To evaluate a novel two-stage technique to increase yield of bacteria isolated from infected corneal ulcers. METHODS: A new blade was designed to remove friable material from infected corneal ulcers. The new blade was used in combination with standard tissue micro-homogenisation equipment in a two-stage technique intended to distribute biopsy samples evenly between relevant agar plates. Patients with presumed-bacterial corneal ulcers underwent sequential corneal sampling using the new two-stage technique and a scalpel blade, used without micro-homogenisation (the order of sampling was varied between two groups). Bacterial isolation rates were compared using the chi-squared test. RESULTS: Twenty-four patients with presumed-bacterial corneal ulcers were studied. The overall positive bacterial isolation rate was 88%, with identical bacterial isolation rates for the new two-stage technique and the scalpel blade (71%). The new technique isolated bacteria from three ulcers that had initially been 'sterile' when sampled with a scalpel blade. Polymicrobial infections were identified in two ulcers with the new blade where only a single organism had been identified using the scalpel blade (not significantly different). CONCLUSIONS: The new two-stage technique shows promise for improving bacterial isolation rates from presumed-bacterial corneal ulcers. PMID- 10692929 TI - Corneal topography in patients with congenital ptosis. AB - PURPOSE: To determine the effect of congenital ptosis on corneal shape, and to assess the role of these topographic changes in the development of amblyopia. METHODS: Twenty-two patients with congenital ptosis were examined and a corneal topographic examination performed in both the ptotic and normal eyes. The qualitative corneal classification was done according to the colour-coded topographic maps. The surface regularity index (SRI) and the surface asymmetry index (SAI) were used as quantitative descriptors of the study. RESULTS: Ptotic eyes had an increased incidence of astigmatism, bow tie pattern on corneal topography, corneal asymmetry (SAI, p < 0.05) and corneal irregularity (SRI, p < 0.05). Lack of mirror-image symmetry with the fellow eye was higher in amblyopic eyes. CONCLUSION: Eyes with congenital ptosis have an increased incidence of astigmatism and a bow tie pattern on corneal topography. These features are associated with the presence of amblyopia. PMID- 10692931 TI - Per-operative retinoscopy as a predictor of final post-operative refraction. AB - PURPOSE: To assess the accuracy of streak retinoscopy performed at the end of cataract surgery as a predictor of final post-operative error. METHOD: Retinoscopy was performed on 68 patients as they lay on the operating table after routine cataract extraction and intraocular lens implantation. In each case the predicted post-operative refraction by biometry and the retinoscopy at the end of the operation were compared with the 6 week post-operative subjective refraction. RESULTS: The retinoscopy had a mean difference of 0.6 D (standard deviation of 0.5 D). The post-operative refraction predicted by biometric measurements had a mean difference of 1.6 D (standard deviation 0.6 D). When corrected for systematic error, 8% of patients were found to have an error of greater than 2 D as predicted by pre-operative biometry. Prediction by retinoscopy made no error greater than 2 D. The accuracy in the retinoscopic prediction of post-operative refraction was significantly better than the biometry using the F-test (p = 0.001). CONCLUSION: Retinoscopy at the end of cataract surgery may be a valuable tool to alert the surgeon to an unexpected refractive error. This would enable immediate intraocular lens exchange, if required. PMID- 10692930 TI - Human aqueous and vitreous humour levels of ciprofloxacin following oral and topical administration. AB - PURPOSE: To assess aqueous and vitreous humour ciprofloxacin concentrations following oral and topical administration of ciprofloxacin in patients with non inflamed cornea and an intact crystalline lens, and to compare the concentrations of the drug given by either route. METHODS: In this prospective study, 34 patients undergoing pars plana vitrectomy for various ocular pathologies were divided into two groups. Eighteen patients received 2 drops of 0.3% ophthalmic solution of ciprofloxacin every 30 min for 3 h and then every 60 min for the next 3 h, and 16 patients received a single oral dose of 1000 mg ciprofloxacin 6 h before surgery. The aqueous and vitreous humour samples were simultaneously harvested after oral or topical administration during pars plana vitrectomy to assess penetration of the drug. These samples were assayed for ciprofloxacin concentrations by a method described previously by us using high-performance liquid chromatography. RESULTS: The aqueous and vitreous humour levels of ciprofloxacin were 0.59 +/- 0.06 microgram/ml (mean +/- SEM) and 0.64 +/- 0.06 microgram/ml after oral and 0.44 +/- 0.07 microgram/ml and 0.22 +/- 0.04 microgram/ml after topical ciprofloxacin administration, respectively. Aqueous humour levels were not statistically significantly different following oral and topical administration (p = 0.069). However, the vitreous level of the drug after oral administration was significantly higher than that after topical administration (p < 0.001). CONCLUSION: Ocular bioavailability of ciprofloxacin in aqueous humour following oral and topical administration is found to be similar when the drug was applied as described above. Penetration of ciprofloxacin into vitreous humour is less than that into aqueous humour after topical administration. PMID- 10692932 TI - Predictive value of family data for the management of infantile bilateral partial cataract. AB - PURPOSE: To examine the data on outcome of surgery performed over a wide age range in members of a family affected by familial infantile bilateral partial cataract, with the purpose of assessing their predictive value concerning the timing of surgery. METHODS: A retrospective clinical study was carried out of a family with dominant inheritance of familial infantile bilateral partial cataract. The family spanned four generations and consisted of 53 members, 31 of whom were examined in our department. Of these, 18 were affected. Cataract surgery was performed in 26 eyes of 15 patients, whose ages ranged from 6 to 58 years at the time of operation. As the surgical procedures spanned the years from 1978 to 1996, different techniques were used. RESULTS: In 24 eyes (92%) the post operative visual acuity was 6/9 or better. One eye achieved 6/12 and another 6/15. CONCLUSIONS: In this particular family there was no relationship between the post-operative visual acuity and the age at which surgery was performed. In deciding when to operate on family members with infantile bilateral partial cataract with similar morphology, in addition to the commonly used criteria the family data should also be taken into account. In infants and young children, delaying surgery may allow better development of visual acuity aided by accommodation, stabilisation of binocularity and more precise determination of the power of an intraocular lens. Success of very early surgery in such cases may not be attributable to the timing of the operation. PMID- 10692933 TI - The results of adjustable suture technique in paediatric strabismus surgery. AB - PURPOSE: Adjustable suture technique (AST) has been shown to be an effective treatment method in adult strabismus. The application of AST is not well studied in children due to potential poor cooperation during adjustment and the concern that the adjustment process can not be completed. The present series evaluates the efficacy and safety of this technique in a preselected group of children between 7 and 15 years of age. METHODS: A retrospective review of 89 consecutive children undergoing AST was completed to assess: (1) the ability to perform and complete adjustment in children; (2) the frequency of need to perform adjustment; and (3) accuracy of surgical alignment. Only patients with horizontal rectus muscle surgery were included. RESULTS: All children successfully completed the AST on the first post-operative day. Of the 89 patients, 24 (27%) required further adjustment by the AST. The mean follow-up period was 13.1 months. Fifty three patients (60%) had previous strabismus surgery (range 1 to 5 operations). Sixty-six (74%) patients achieved successful alignment. Complications included slipped muscles in 1 case and difficulty in recession of the lateral rectus muscle in 1 patient. CONCLUSIONS: Twenty-seven per cent of the patients required post-operative adjustment. The AST achieved an overall 74% successful alignment. Application of the AST should be considered in children with horizontal deviations over age 7 years, especially in the reoperations of esotropia. PMID- 10692934 TI - Transscleral diode laser in the treatment of retinopathy of prematurity. AB - PURPOSE: To assess the outcome of contact transscleral diode laser (TSDL) in the treatment of threshold retinopathy of prematurity (ROP). METHOD: TSDL was performed in 14 eyes of 8 babies who presented to the paediatric ophthalmic service at King's College Hospital between 1996 and 1997 with threshold ROP (median post-conceptual age 26 + 1 weeks, median birthweight 835 g) by a single surgeon. Follow-up ranged from 9 to 41 weeks (median 21 weeks). RESULTS: In 11 eyes (79%) regression of ROP occurred after a single laser treatment with a good anatomical outcome. In 3 eyes (21%) there was an unfavourable response with the development of traction retinal detachment. These include both eyes of one baby who rapidly progressed to stage IV ROP. One other eye developed a fibrotic band a few months after treatment. No significant complications of laser treatment were observed. CONCLUSION: These initial results indicate that TSDL photocoagulation is an effective and technically straightforward alternative to cryotherapy in the treatment of ROP. PMID- 10692935 TI - Blue light induced apoptosis in rat retina. AB - PURPOSE: To explore cell death in blue light induced retinal damage. METHODS: Sprague-Dawley rats reared under cyclic light were exposed continuously to diffuse blue light (400-480 nm) at 0.64 W/m2 for 3 or 6 h after 22 h of dark adaptation. The rats were kept in darkness and killed immediately, 8, 16 and 24 h following light exposure. The retinal damage by the blue light was examined with a transmission electron microscope. The cell death was characterised by in situ terminal dUTP nick end labelling (TUNEL) and gel electrophoresis. RESULTS: During the 24 h following light exposure, photoreceptor cell death was characterised by progressive condensation and margination of the chromatin, shrinkage or convolution and fragmentation of the nucleus, condensation of the cytoplasm, and formation of apoptotic bodies along with rapid removal of dying cells from damaged areas in the absence of inflammatory response. The TUNEL-positive nuclei were scattered individually in the outer nuclear layer just after light exposure. A wave of massive TUNEL labelling of photoreceptor nuclei peaked at 8-16 h and dropped at 24 h following light exposure. The distribution of TUNEL-positive nuclei was located predominantly at the upper temporal region of the retina, which was the most sensitive area to the damage caused by blue light. Furthermore, the multiples of internucleosomal cleavage of 180-200 base pairs were demonstrated at corresponding time points. CONCLUSION: Photoreceptor cell apoptosis is seen early after the retina is damaged by blue light. PMID- 10692936 TI - The use of combined intravenous pulse methylprednisolone and low-dose oral cyclosporin A in the treatment of corneal graft rejection: addendum to previous report. PMID- 10692937 TI - Hypopyon keratitis in corneal epithelial basement membrane dystrophy. PMID- 10692938 TI - Bilateral keratomalacia in a cachectic scleroderma patient. PMID- 10692939 TI - Acanthamoeba keratitis following optical keratoplasty. PMID- 10692940 TI - Mucocele arising from a lateral extension of a sphenoid sinus. PMID- 10692941 TI - Aberrant lacrimal gland within the tarsal plate presenting as hyperlacrimation. PMID- 10692942 TI - Desmoplastic trichilemmoma of the upper eyelid. PMID- 10692943 TI - An unusual chorioretinal dystrophy? PMID- 10692944 TI - Mitochondrial DNA disease masquerading as age-related mascular degeneration. PMID- 10692945 TI - Candida endophthalmitis: a diagnostic dilemma. PMID- 10692946 TI - Ocular Munchausen syndrome characterised by self-introduction of chalk concretions into the conjunctival fornix. PMID- 10692947 TI - An unusual presentation of Graves' disease. PMID- 10692948 TI - Assessment of choroidal involvement in sarcoidosis with indocyanine green angiography. PMID- 10692949 TI - Intravitreal penetration of teicoplanin. PMID- 10692950 TI - Primary empty sella: cause of visual failure or chance association? PMID- 10692951 TI - Glaucoma screening by optometrists: positive predictive value of visual field testing. PMID- 10692952 TI - Post-operative myopic shift due to trapped intracapsular Healon. PMID- 10692953 TI - Challenges to public health in the new millennium. PMID- 10692954 TI - Drinking water and gastrointestinal disease: need of better understanding and an improvement in public health surveillance. PMID- 10692955 TI - The unseen face of humanitarian crisis in eastern Democratic Republic of Congo: was nutritional relief properly targeted? AB - STUDY OBJECTIVE: Comparison of children's nutritional status in refugee populations with that of local host populations, one year after outbreak refugee crisis in the North Kivu region of Democratic Republic of Congo. DESIGN: Cross sectional surveys. SETTING: Temporary and other settlements, in the town of Goma and surrounding rural areas. SUBJECTS: Anthropometric indicators of nutritional status and presence or absence of oedema were measured among 5121 children aged 6 to 59 months recruited by cluster sampling with probability proportional to size, between June and August 1995. RESULTS: Children in all locations demonstrated a typical pattern of growth deficit relative to international reference. Prevalence of acute malnutrition (wt/ht < -2 Z score) was higher among children in the rural non-refugee populations (3.8 and 5.8%) than among those in the urban non-refugee populations (1.4%) or in the refugee population living in temporary settlements (1.7%). Presence of oedema was scarcely noticed in camps (0.4%) while it was a common observation at least in the most remote rural areas (10.1%). As compared with baseline data collected in 1989, there is evidence that nutritional status was worsening in rural non-refugee populations. CONCLUSIONS: Children living in the main town or in the refugee camps benefited the most from nutritional relief while those in the rural non-refugee areas were ignored. This is a worrying case of inequity in nutritional relief. PMID- 10692956 TI - Women in hospital medicine in the United Kingdom: glass ceiling, preference, prejudice or cohort effect? AB - OBJECTIVE: To assess from official statistics whether there is evidence that the careers of women doctors in hospitals do not progress in the same way as those of men. DESIGN: The proportions of female hospital doctors overall (1963-96), and in the specialties of medicine, surgery, obstetrics and gynaecology, pathology, radiology/radiotherapy, anaesthetics and psychiatry (1974-1996) were examined. Additionally data were examined on career preferences and intentions from pre registration house officers, final year medical students, and medical school applicants (1966-1991). ANALYSIS: Data were analysed according to cohort of entry to medical school to assess the extent of disproportionate promotion. RESULTS: The proportion of women in hospital career posts was largely explained by the rapidly increasing proportion of women entering medical school during the past three decades. In general there was little evidence for disproportionate promotion of women in hospital careers, although in surgery, hospital medicine and obstetrics and gynaecology, fewer women seemed to progress beyond the SHO grade, and in anaesthetics there were deficits of women at each career stage. Analyses of career preferences and intentions suggest that disproportionate promotion cannot readily be explained as differential choice by women. CONCLUSIONS: Although there is no evidence as such of a "glass ceiling" for women doctors in hospital careers, and the current paucity of women consultants primarily reflects historical trends in the numbers of women entering medical school, there is evidence in some cases of disproportionate promotion that is best interpreted as direct or indirect discrimination. PMID- 10692957 TI - Nutrient intakes during pregnancy: the influence of smoking status and age. AB - STUDY OBJECTIVE: To examine the relation of antioxidant and other nutrient intakes in pregnancy to smoking and sociodemographic variables. DESIGN: Cohort study. SETTING: St Mary's Maternity Hospital, Portsmouth. PARTICIPANTS: Pregnant nulliparous women, with no existing complications of pregnancy, were recruited from antenatal booking clinics. A total of 774 women completed seven day food diaries, and supplied detailed data on their use of nutrient supplements. MAIN RESULTS: Smokers had lower intakes of most micronutrients. After adjustment for the confounding effects of maternal age, height, and education, only vitamin C and carotenoid intakes remained significantly depressed. Age was strongly and significantly associated with the intake of most nutrients, including antioxidants, and this association was independent of other maternal factors. Antioxidant intake was therefore lowest in young women who smoked: for example smokers under 24 years had a mean vitamin C intake of 57 mg (SD 35) compared with 106 mg (SD 52) for non-smokers aged 28 and over (difference 49 mg, 95% CI 39, 59). The corresponding intakes of carotenoid equivalents were 1335 micrograms (SD 982) and 2093 micrograms (SD 1283) (difference 758 micrograms, 95% CI 496, 1020). CONCLUSIONS: The study has identified, for the first time, young pregnant women as a group at particular risk of low micronutrient intake. The health implications of poor nutrition now need to be evaluated, particularly for those women who smoke. PMID- 10692958 TI - Social inequalities in health related behaviours in Barcelona. AB - OBJECTIVE: This study describes social class inequalities in health related behaviours (tobacco and alcohol consumption, physical activity) among a sample of general population over 14 years old in Barcelona. DESIGN: Cross sectional study (Barcelona Health Interview Survey). SETTING: Barcelona city (Spain). PARTICIPANTS: A representative stratified sample of the non-institutionalised population resident in Barcelona was obtained. This study refers to the 4171 respondents aged over 14. DATA: Social class was obtained from a Spanish adaptation of the British Registrar General classification. In addition, sociodemographic variables such as family structure and employment status were used. As health related behaviours tobacco consumption, alcohol consumption, usual physical activity and leisure time physical activity were analysed. Age adjusted percentages were compared by social class. Multivariate analysis was performed using logistic regression models. MAIN RESULTS: Women in the upper social classes were more likely to smoke, the adjusted odds ratio (OR) for social class V in reference to social class I was 0.36 (95% confidence intervals (95%CI): 0.19, 0.67), while the opposite occurred among men although it was not statistically significant in multivariate analysis. Smoking cessation was more likely among men in the higher classes (OR for class V 0.41, 95%CI: 0.18, 0.90). Excessive alcohol consumption among men showed no differences between classes, while among women it was greater in the upper classes. Engaging in usual physical activity classified as "light or none" in men decreased with lowering social class (OR class IVa: 0.55 and OR class IVb: 0.47). Women of social classes IV and V were less likely to have two or more health risk behaviours (OR for class V 0.33, 95% CI: 0.18, 0.62). CONCLUSION: Health damaging behaviours are differentially distributed among social classes in Barcelona. Health policies should take into account these inequalities. PMID- 10692959 TI - Homocysteine, folate, vitamin B12, and cardiovascular risk in Indians, Malays, and Chinese in Singapore. AB - OBJECTIVE: To examine the hypothesis that the higher rates of coronary heart disease (CHD) in Indians (South Asians) compared with Malays and Chinese is partly attributable to differences in blood concentrations of homocysteine, and related blood concentrations of folate and vitamin B12. DESIGN: Cross sectional study of the general population. SETTING: Singapore. PARTICIPANTS: Random sample of 726 fasting subjects aged 30 to 69 years. MAIN RESULTS: Mean plasma total homocysteine concentrations did not show significant ethnic differences; values were Indians (men 16.2 and women 11.5 mumol/l), Malays (men 15.0 and women 12.5 mumol/l), and Chinese (men 15.3 and women 12.2 mumol/l). Similarly, the proportions with high plasma homocysteine (> 14.0 mumol/l) showed no important ethnic differences being, Indians (men 60.0 and women 21.9%), Malays (men 53.9 and women 37.8%), and Chinese (men 56.6 and women 30.6%). Mean plasma folate concentrations were lower in Indians (men 8.7 and women 10.9 nmol/l) and Malays (men 8.5 and women 10.8 nmol/l), than Chinese (men 9.7 and women 13.8 nmol/l). Similarly, the proportions with low plasma folate (< 6.8 nmol/l) were higher in Indians (men 44.9 and women 36.6%) and Malays (men 45.3 and women 24.5%) than Chinese (men 31.4 and women 12.6%). Mean plasma vitamin B12 concentrations were lowest in Indians (men 352.5 and women 350.7 pmol/l), then Chinese (men 371.1 and women 373.7 pmol/l), and then Malays (men 430.5 and women 486.0 pmol/l). CONCLUSION: While there were ethnic differences for plasma folate and vitamin B12 (in particular lower levels in Indians), there was no evidence that homocysteine plays any part in the differential ethnic risk from CHD in Singapore and in particular the increased susceptibility of Indians to the disease. PMID- 10692960 TI - Impairments, disabilities and needs assessment among non-fatal war injuries in south Lebanon, Grapes of Wrath, 1996. AB - STUDY OBJECTIVE: To examine the impact of non-fatal war related injuries on physical disability in a group of war wounded civilians and to assess their needs. DESIGN: Cross sectional study. Home interviews were conducted using a structured interview schedule around one month after the injury, to assess impairments, disabilities, and needs. STUDY POPULATION AND SETTING: War wounded persons in towns and villages in South Lebanon during the attack "Grapes of Wrath" in 1996. RESULTS: The majority of the study population were young and in their productive age, mostly injured in the street or while hiding in open shelters. Around half of the injuries resulted in impairments, but, there were no age, gender or geographical differentials by severity of impairment. Almost one third (29%) of the students enrolled in schools at the time of the injury reported failure to continue their education and 42% of the working members lost their jobs with no potential for 34% of them to resume their former jobs. The impact of the injury on impairments, motor disabilities and physical independence was highest for injuries to the lower limbs (age and sex adjusted risk ratio (RR) 1.62, 95% confidence intervals (CI) 1.25, 2.10; 2.98, 95% CI 2.09, 4.23; and 2.13, 95% CI 1.39, 3.27, respectively). Despite the acute and early relief services provided by all those concerned at the time of the injury, when asked about unmet needs, the majority of the impaired (66%) reported the need for additional services, mostly medical in nature. The degree of disability was a salient factor for the need for rehabilitative services but not for medical services. CONCLUSIONS: The chronic and diverse needs of people with war injuries are often neglected and underestimated by the governmental institutions and relief agencies. Research funds as well as services should be allocated to tackle the long term and continuous health and social needs of those injured and their families. PMID- 10692961 TI - Empowering the deaf. Let the deaf be deaf. AB - Deafness is often regarded as just a one and only phenomenon. Accordingly, deaf people are pictured as a unified body of people who share a single problem. From a medical point of view, we find it usual to work with a classification of deafness in which pathologies attributable to an inner ear disorder are segregated from pathologies attributable to an outer/middle ear disorder. Medical intervention is thus concerned more with the origin, degree, type of loss, onset, and structural pathology of deafness than with communicative disability and the implications there may be for the patient (mainly dependency, denial of abnormal hearing behaviour, low self esteem, rejection of the prosthetic help, and the breakdown of social relationships). In this paper, we argue that hearing loss is a very complex phenomenon, which has many and serious consequences for people and involves many factors and issues that should be carefully examined. The immediate consequence of deafness is a breakdown in communication whereby the communicative function needs to be either initiated or restored. In that sense, empowering strategies--aimed at promoting not only a more traditional psychological empowerment but also a community one--should primarily focus on the removal of communication barriers. PMID- 10692962 TI - Drinking water turbidity and gastrointestinal illness in the elderly of Philadelphia. AB - STUDY OBJECTIVE: To investigate the association between drinking water quality and gastrointestinal illness in the elderly of Philadelphia. DESIGN: Within the general population, children and the elderly are at highest risk for gastrointestinal disease. This study investigates the potential association between daily fluctuations in drinking water turbidity and subsequent hospital admissions for gastrointestinal illness of elderly persons, controlling for time trends, seasonal patterns, and temperature using Poisson regression analysis. SETTING AND PARTICIPANTS: All residents of Philadelphia aged 65 and older in 1992 1993 were studied through their MEDICARE records. MAIN RESULTS: For Philadelphia's population aged 65 and older, we found water quality 9 to 11 days before the visit was associated with hospital admissions for gastrointestinal illness, with an interquartile range increase in turbidity being associated with a 9% increase (95% CI 5.3%, 12.7%). In the Belmont service area, there was also an association evident at a lag of 4 to 6 days (9.1% increase, 95% CI 5.2, 13.3). Both associations were stronger in those over 75 than in the population aged 65 74. This association occurred in a filtered water supply in compliance with US standards. CONCLUSIONS: Elderly residents of Philadelphia remain at risk of waterborne gastrointestinal illness under current water treatment practices. Hospitalisations represent a very small percentage of total morbidity. PMID- 10692963 TI - The Communicable Disease Surveillance System in the Kosovar refugee camps in the former Yugoslav Republic of Macedonia April-August 1999. PMID- 10692964 TI - Morbidity and health care utilisation among elderly people in Mmankgodi village, Botswana. AB - OBJECTIVE: To evaluate the health status among the elderly in a village in Botswana and their pattern of health care utilisation. DESIGN: A descriptive study where all persons 60 years and older were invited to participate, including a medical examination, laboratory testing and a questionnaire aiming at gathering sociodemographic data. SETTING: Mmankgodi village of Botswana. SUBJECTS: 419 persons were identified as elderly in the village, out of which 337 were included. MAIN OUTCOME MEASURES: The general medical examination also included eye status, vision and hearing tests, nutritional status, blood pressure and registering of physical disabilities. Laboratory tests included haemoglobin, blood glucose, HIV antibodies and serum lipids. The questionnaire contained questions regarding family and civil status, self assessed general health, health problems experienced during the previous month, and health care utilisation. Questions also pertained to smoking, taking snuff, and alcohol consumption. RESULTS: A majority (75%) of the elderly experienced good or only somewhat reduced health, while one quarter suffered more serious health problems. The most frequent health problems were related to the musculoskeletal system. Eye diseases, including cataract and blindness, were also common. The concentration of serum lipids is lower than the one found in the elderly population of Norway. Nutritional status indicated a relatively high prevalence (7%) of malnutrition. The majority of men were still married (87%), while most women were widowed (71%). Women reported more health problems than men, and they also reported more worries regarding their own life situation. There is a tendency for the elderly to seek assistance from the established clinics and other health facilities for their health problems. Worries are either kept to themselves or advice is sought from relatives. Traditional healers were not often consulted for health problems or worries. CONCLUSIONS: Major health problems were identified among the elderly in this geographical area of Botswana. There is presently no health programme in Botswana aimed at the elderly. Some of the diseases and conditions found in this study could easily be identified and treated in the present health system through a health care programme. PMID- 10692965 TI - How does tuberculosis relate to HIV positive and HIV negative drug users? AB - OBJECTIVES: (1) To compare the incidence of active tuberculosis in HIV positive and HIV negative drug users. (2) To describe the main characteristics of the tuberculosis cases. DESIGN: A prospective study was performed from 1986 to 1996 as part of an ongoing cohort study of HIV infection in Amsterdam drug users. METHODS: Data from the cohort study, including HIV serostatus and CD4-cell numbers, were completed with data from the tuberculosis registration of the tuberculosis department of the Amsterdam Municipal Health Service. Analyses were carried out with person time and survival methods. RESULTS: Of 872 participants, 24 persons developed culture confirmed tuberculosis during a total follow up period of 4000 person years (0.60 per 100 py, 95% CI: 0.40, 0.90). Nineteen cases were HIV positive (1.54 per 100 py, 95% CI: 0.86, 2.11) and five HIV negative (0.18 per 100 py, 95% CI: 0.08, 0.43). Multivariately HIV infection (relative risk: 12.9; 95% CI: 3.4, 48.8) and age above 33 years (RR: 6.8; 95% CI: 1.3, 35.0, as compared with age below 27) increased the risk for tuberculosis substantially. Additional findings were: (1) 13 of 22 pulmonary tuberculosis cases (59%) were detected by half yearly radiographic screening of the chest; (2) tuberculosis occurred relatively early in the course of HIV infection at a mean CD4 cell number of 390/microliter; (3) an estimated two thirds of the incidence of tuberculosis observed among HIV positive cases was caused by reactivation; (4) all but one patient completed the tuberculosis treatment. CONCLUSION: HIV infection increases the risk for active tuberculosis in Amsterdam drug users 13 fold. The incidence of tuberculosis in HIV negative drug users is still six times higher than in the overall Amsterdam population. In the absence of contact tracing and screening with tuberculin skin tests, periodic chest radiographic screening contributes substantially to early casefinding of active tuberculosis in Amsterdam drug users. PMID- 10692966 TI - Southern Oscillation Index and transmission of the Barmah Forest virus infection in Queensland, Australia. PMID- 10692967 TI - The effect of a direct payment or a lottery on questionnaire response rates: a randomised controlled trial. PMID- 10692968 TI - Air pollution and mortality in a cohort of patients with chronic obstructive pulmonary disease: a time series analysis. PMID- 10692969 TI - Hospital admissions for asthma and chronic obstructive airways disease in east London hospitals and proximity of residence to main roads. PMID- 10692970 TI - Non-fatal head injury among Scottish young people: the importance of assault. PMID- 10692971 TI - Treatment with ivermectin: what works in one community may not work in another. PMID- 10692972 TI - Interim report and recommendations of the World Health Organization Task-Force for Osteoporosis. PMID- 10692973 TI - Bone mineral density and vertebral fractures in men. AB - In women, many studies indicate that the risk of vertebral fragility fractures increases as bone mineral density (BMD) declines. In contrast, few studies are available for BMD and vertebral fractures in men. It is uncertain that the strength of the relationship between BMD and fractures is similar in magnitude in middle-aged men and in postmenopausal women. In the present study, 200 men (mean age 54.7 years) with lumbar osteopenia (T-score < -1.5) were recruited to examine the relationships between spine BMD and hip BMD and the associations of BMD with vertebral fractures. Lumbar BMD was assessed from L2 to L4, in the anteroposterior view, using dual-energy X-ray densitometry. At the upper left femur, hip BMD was measured at five regions of interest: femoral neck, trochanter, intertrochanter, Ward's triangle and total hip. Spinal radiographs were analyzed independently by two trained investigators and vertebral fracture was defined as a reduction of at least 20% in the anterior, middle or posterior vertebral height. Spinal radiographs evidenced at least one vertebral crush fracture in 119 patients (59.5%). The results of logistic regression showed that age, femoral and spine BMDs were significant predictors of the presence of a vertebral fracture. Odds ratios for a decrease of 1 standard deviation ranged from 1.8 (1.3-2.8) for spine BMD to 2.3 (1.5-3.6) for total hip BMD. For multiple fractures odds ratios ranged from 1.7 (1.1-2.5) for spine BMD to 2.6 (1.7-4.3) for total hip BMD. In all models, odds ratios were higher for hip BMD than for spine BMD, particularly in younger men, under 50 years. A T-score < -2.5 in the femur (total femoral site) was associated with a 2.7-fold increase in the risk of vertebral fracture while a T-score < -2.5 in the spine was associated with only a 2-fold increase in risk. This study confirms the strong association of age and BMD with vertebral fractures in middle-aged men, shows that the femoral area is the best site of BMD measurement and suggests that a low femoral BMD could be considered as an index of severity in young men with lumbar osteopenia. PMID- 10692974 TI - Influence of heredity and environment on bone density in adolescent boys: a parent-offspring study. AB - The purpose of the present parent-offspring study was to investigate the influence of heredity and environment on bone density in young men. Another aim was to discover whether the same genetic factors influence bone mass, lean mass and muscle strength. Fifty families including a father, mother and one son were investigated. The mothers (aged 44.5 +/- 4.4 years) and fathers (aged 47.1 +/- 4.4 years) generally had a sedentary lifestyle with little physical activity. As a contrast, all but three of the sons (aged 17.0 +/- 0.4 years) were active in ice hockey training. Bone mineral density (BMD, g/cm2) of the total body, head, lumbar spine and femoral neck was measured using dual-energy X-ray absorptiometry. Muscle strength of the hamstrings and quadriceps muscles was also measured in the boys. BMD values of different sites in the fathers, mothers and sons were adjusted for weight, height, age, and any significant influence of environment. Heritability estimates were obtained as regression coefficients with the boys' adjusted BMD as dependent variable and the adjusted midparent bone density (father BMD + mother BMD/2) as independent variable. Accordingly, heritability explained 34-54% of the variation in the sons' BMD. Midparent BMD of several sites also predicted the boys' lean mass and quadriceps strength, and midparent-offspring differences in lean mass predicted midparent-offspring differences in BMD of the total body, head and spine (beta = 0.30-0.51, p < 0.05). The sons were found to have almost 30% higher femoral neck BMD than their fathers, and physical activity (hours/week) predicted BMD at several sites among the sons beta = 0.26-0.34, p < 0.05). In conclusion, heritability is a main determinant of the variance in BMD in young men. Based on the results we suggest that the same genetic factors may influence bone mass, lean mass and muscle strength by affecting body size. The present study also emphasizes the importance of physical activity for the development and maintenance of BMD in men. PMID- 10692975 TI - Efficiency of quantitative ultrasound measurements as compared with dual-energy X ray absorptiometry in the assessment of corticosteroid-induced bone impairment. AB - Bone loss due to corticosteroid treatment differs from that of postmenopausal osteoporosis with regard to bone structure. Corticosteroids affect both horizontal and vertical trabeculae while horizontal trabeculae are damaged in postmenopausal osteoporosis. Dual-energy X-ray absorptiometry (DXA) is the gold standard to evaluate bone loss. The place of quantitative ultrasound (QUS), a technique that could theoretically provide information on bone structure, is not well established in corticosteroid-induced bone impairment. The aim of the study was to determine the usefulness of QUS in the assessment of corticosteroid induced bone impairment. We hypothesized that the relationship between QUS and DXA could be influenced by changes in bone structure and thus differ with regard to corticosteroid treatment. Seventy-seven women with inflammatory diseases chronically treated with corticosteroids (dose: 7.5-15 mg/day), 29 without corticosteroids and 100 controls were investigated. Bone mineral density at the lumbar spine (BMDL) was measured by DXA and QUS parameters were measured at the calcaneus. Both the QUS parameters (SOS, BUA, Stiffness) and BMDL were significantly lower (by 1.3% for SOS, 5.8% for BUA, 12.7% for Stiffness and 11% for BMDL) in patients treated with corticosteroids compared with patients not taking corticosteroids and with controls (p < 0.001, ANCOVA, with age and height as covariates). Multiple linear regressions of Stiffness, SOS and BUA as dependent variables on age, BMDL, corticosteroid treatment and a computed new variable designed to test the interaction between BMDL and the treatment group showed that Stiffness, SOS and BUA were dependent on age and BMDL (p < 0.001); BUA and Stiffness were dependent on treatment group. Taking into account the age of the patients, a significant difference was observed in the relation between BUA and BMDL according to treatment with corticosteroids. A similar difference was found in the subgroup of patients without fractures. SOS and BUA were strongly correlated but their relation did not differ according to treatment. Thus, QUS is useful in the assessment of corticosteroid-associated bone loss. Furthermore, the observation of a significant difference in the relationship between BUA and BMDL with regard to corticosteroid treatment might support the hypothesis that QUS, especially BUA, could give additional information about bone structure. PMID- 10692976 TI - Treatment of reduced bone mineral density in athletic amenorrhea: a pilot study. AB - There is considerable concern about the adverse effects on the skeleton of loss of menstrual function as a result of athletic activity, as well as uncertainty as to how it should be managed clinically. In a pilot intervention study 34 elite middle and long-distance runners, aged 18-35 years, with menstrual irregularity due to their athletic activity were randomized to three groups: (A) to receive hormone replacement therapy (HRT) and 1000 mg calcium per day (n = 10), (B) to receive 1000 mg calcium per day (n = 14), (C) a control group who received no treatment (n = 10). Bone mineral density (BMD) was measured in the left hip and lumbar spine (L2-4) using dual-energy X-ray absorptiometry. Results were first analyzed according to whether menstruation returned, either naturally or secondary to HRT (EU), and compared with those from subjects who remained amenorrheic (AM). During the first year BMD increased in the EU group in Ward's triangle (3.8%) and the lumbar spine (4.1%; both P < 0.05). BMD fell in the AM group in all regions and the between-group differences were 5.6% (p < 0.02) in Ward's triangle, 5.8% (p < 0.02) in L2-4 and 3.9% in the trochanter (p < 0.05). An 'intention to treat' analysis was then performed. It was found that the mean relative improvement at 1 year in spinal BMD was only 1.5%, due to return of menses in some of the controls and withdrawals from treatment in the treatment group. In consequence, a trial designed to show, with 80% power and 5% significance, a measurable benefit in lumbar spine BMD resulting from allocation to HRT treatment would require about 1150 athletes with amenorrhea or oligomenorrhea. These numbers could be reduced substantially to 380 subjects by confining the trial to completely amenorrheic athletes, who in this study were less likely to regain menses. For these and other logistical reasons, an HRT trial in amenorrheic athletes could only be successfully organized through international collaboration. This study illustrates the major effects of treatment withdrawals and instability of menstrual status on the design of longitudinal studies on the bony effects of menstrual dysfunction prior to menopause. PMID- 10692977 TI - Premature greying of the hair is not associated with low bone mineral density. AB - In two recent case-control studies premature greying of the hair was associated with a lowering of bone mineral density (BMD) and osteopenia, suggesting that this might be a clinically useful risk marker for osteoporosis. We report a further re-examination of this proposal in 52 prematurely grey-haired women from East Yorkshire who responded to an advertisement inviting them for bone densitometry. Thirty-five had no clinical or drug history that could influence bone density. All were Caucasian with a mean age of 52.8 years. In the group as a whole the mean BMD values at the lumbar spine and femoral neck were no different from those of a young adult, but there was a trend toward a greater than average BMD than that of the local age-matched population (p = 0.097 and 0.218, respectively). Twenty women were premenopausal, with an average age of 45.3 years. Mean BMD values at the lumbar spine and femoral neck in this group were no different from those of young adults. There was, however, a trend toward a BMD greater than that of the local age-matched population at the femoral neck (p = 0.117). Fifteen women were postmenopausal with an average age of 62.9 years and an average age at menopause of 51.1 years. Mean BMD values at both the lumbar spine and femoral neck in this group were lower than those of young adults, but no different from those of the local age-matched population. In conclusion, our group of prematurely grey-haired women had average BMD for their age, and we are therefore unable to support the proposed clinical usefulness of premature greying as a risk marker for osteoporosis. PMID- 10692978 TI - Disproportionate, age-related bone loss in long bone ends: a structural analysis based on dual-energy X-ray absorptiometry. AB - The width of long bone diaphyses apparently increase with age, a phenomenon that is suggested to have some positive impact on bone strength. On the other hand, these changes in size that are site-specific may cause a deterioration in the local mechanical integrity of the whole bone. Physical activity and calcium intake are known to be able to modify bone mass and size. It is, however, not known whether these lifestyle habits can modify the postulated disproportionate changes in bone size. To address this question, bone mineral content (BMC) derived estimates of cross-sectional areas (CSA) of femur and radius in 158 premenopausal (mean age 43, standard deviation 2 years) and 134 postmenopausal (63 (2) years), clinically healthy women with contrasting long-term histories in physical activity and calcium intake were determined from dual-energy X-ray absorptiometry (DXA) data. The DXA-obtained BMC correlated strongly with the actual CSA (r = 0.94) determined with peripheral quantitative computed tomography. The ratios between functionally interrelated CSA data (i.e., (radial shaft CSA/distal radius CSA), (trochanter CSA/femoral neck CSA), (femoral shaft CSA/trochanter CSA) and (femoral shaft CSA/femoral neck CSA)) were considered primary outcome variables. Neither physical activity nor calcium intake separately or interactively were associated with any CSA ratio. Age showed no interaction with physical activity or calcium intake but was independently associated with all CSA ratios, except the ratio of femoral shaft CSA to trochanteric CSA. This study indicated clearly that a preferential reduction in the cross-sectional area occupied by bone mineral occurs disproportionately at the long bone ends as compared with diaphyseal sites, and this apparently inherent, age-associated relative loss seems not to be prevented by physical activity or calcium intake. In particular, given the utmost clinical relevance of the proximal femur region, an observed loss in femoral neck CSA of about 10% in contrast to about a 5% loss in trochanteric CSA warrants further investigation regarding its potential role as a predictor for hip fracture. Not only the local differences in bone composition but also the biomechanical aspects are important factors underlying these apparent changes in CSA at the studied skeletal sites. PMID- 10692979 TI - VDR genotype and response to etidronate therapy in late postmenopausal women. AB - Twenty-four late postmenopausal women with osteoporosis were studied. The patients were separated in three subgroups according to the BsmI polymorphism of the vitamin D receptor (VDR) gene: BB (n = 8), Bb (n = 10) and bb (n = 6). They did not differ in age (mean ages were 66.0 years, 65.9 years and 63.9 years, respectively), years after menopause (18.7 years, 18.1 years and 18.4 years) or body weight (64.9 kg, 65.3 kg and 63.8 kg), the variables known to be associated with bone mineral density (BMD). The results show that the response to antiresorptive bisphosphonate therapy in combination with calcium supplementation is modified by VDR genotype. The lumbar spine BMD increased significantly faster in the BB and Bb groups (7.3% and 7.0%, respectively) compared with the bb group (2.5%) during 1 year of cyclic etidronate therapy (400 mg/day) and calcium supplementation (1000 mg/day). The biochemical marker of bone resorption (urinary hydroxyproline excretion) as well as the bone formation marker (serum levels of osteocalcin) decreased during the treatment. With respect to VDR genotype, a significantly higher decrease in osteocalcin level was observed in bb as compared with BB subjects. We conclude that the VDR genotype is involved in an individual's response to cyclic etidronate therapy with calcium supplementation. PMID- 10692980 TI - Bone mineral in pre- and postmenopausal women with insulin-dependent and non insulin-dependent diabetes mellitus. AB - The aim of the study was to compare bone mineral density (BMD) and bone turnover in pre- and postmenopausal women with insulin-dependent diabetes mellitus (IDDM), non-insulin-dependent diabetes mellitus (NIDDM) and normal reference women. In a cross-sectional study 31 and 11 premenopausal and 22 and 21 postmenopausal IDDM and NIDDM patients, respectively, were recruited from an outpatient clinic. BMD in the forearm, spine, femur and total body and biochemical markers of bone turnover were measured and compared with reference values obtained from measurements of normal healthy pre- and postmenopausal women. Postmenopausally, but not premenopausally, IDDM patients had lower BMD values than NIDDM patients. Postmenopausal NIDDM patients had higher BMD value than normal women. The differences in BMD between IDDM and NIDDM patients could be explained statistically by differences in body weight between the NIDMM (obese) and IDDM (lean) women. Markers of bone turnover were significantly higher postmenopausally than premenopausally in both IDDM and NIDDM patients. Osteocalcin was significantly lower in postmenopausal NIDDM compared with postmenopausal IDDM patients and reference values. Otherwise there were no differences in the markers of bone turnover between NIDDM and IDDM patients. In conclusion, postmenopausal IDDM patients have a relatively decreased BMD, whereas NIDDM patients seem to be relatively protected from postmenopausal bone loss. PMID- 10692981 TI - Perception of osteoporosis by Belgian women who work in a university hospital. AB - Campaigns to increase 'awareness' of osteoporosis have been organized. The aim of this study was to assess how Belgian women who benefit from superior conditions favoring 'awareness' perceive osteoporosis as being an important disease. A survey sent to the private home of all the women working in a university hospital in Brussels (n = 1154). From a list of 13 diseases the women were asked to rank, by order of importance, the five which they found to be the most important for a woman of their age. They were also asked about visits to physicians, and screening procedures. The response rate was 55.4%. A high uptake of medical visits and screening procedures was reported: 89% of the women had seen a general practitioner or a gynecologist and 81.6% had undergone at least one gynecologic examination during the previous year. Three times more women had ever undergone mammography than a bone mineral density (BMD) measurement. Overall, 18.1% reported having had a BMD measurement in the past. In women over 50 years, 61% reported having had a BMD measurement and 92.7% having had a mammogram. Osteoporosis was ranked among the five most important diseases by 19.4% of women before the age of 50 years and by 39.3% after that age, far behind breast cancer (respectively 86.3% and 77.7%) and uterine cancer (respectively 74.2% and 58.0%). Thus even among a population of women who benefit from superior conditions for information and screening, the perception of osteoporosis remains low, as does the uptake of osteoporosis screening. PMID- 10692982 TI - Calcaneal ultrasound attenuation in an elderly population: measurement position and relationships with body size and past fractures. AB - This study demonstrates, the relationship between past fracture, body size and broadband ultrasound attenuation (BUA) and investigates two sites of BUA measurement in a representative elderly population of men and women (n = 2106). We measured BUA at a fixed position and at a consistent anatomic position within the calcaneus. We found fixed BUA was less closely correlated with stature and age than anatomic BUA. Both correlations were substantially weaker in men than in women. Mean BUA was significantly lower in women with a past fracture compared with nonfracturers (fixed BUA 63.3 vs 69.4 dB/MHz, p = 0.0004; anatomic BUA 77.6 vs 81.7 dB/MHz, p = 0.013). However, in women, the fixed BUA was better than the anatomic BUA at discriminating between fracturers and nonfracturers (OR 1.38/SD (95% CI 1.12-1.68) and OR 1.22/SD (0.99-1.52), respectively) when adjusted for body size and age. There was no significant difference in either BUA in men with or without a past fracture. In conclusion, currently the fixed position for BUA measurement is preferable and, whilst we have demonstrated that it is possible to locate an anatomically consistent point in the calcaneus, the position chosen by this study did not provide a measurement with more discriminatory capability than the fixed position. In women, BUA behaves similarly to bone mineral density in relation to stature and in its strength of association with past fracture, while the lack of association in men may reflect differing contributions by bone strength to fracture risk in the sexes. PMID- 10692983 TI - Incomplete renal tubular acidosis in 'primary' osteoporosis. AB - Chronic metabolic acidosis may increase alkali mobilization from bone and thus promote the development of osteoporosis. While it is undisputed that overt metabolic acidosis is associated with metabolic bone disease, renal acidification in patients with idiopathic osteoporosis has not been studied systematically. The purpose of this study was to investigate the prevalence of renal acidification defects in patients with 'primary' osteoporosis. Thirty-two women (including 10 premenopausal women) and 16 men who were referred to our department for investigation of osteoporosis were enrolled in this study. Patients with obvious or possible secondary osteoporosis were excluded. None of the patients had overt metabolic acidosis. In random urine samples 12 of the 48 patients had pH levels below 5.5 and were therefore considered to have normal renal acidification. The remaining 36 patients underwent further testing by a short-course oral ammonium chloride load. In this test nine of these 36 patients (7 men and 2 premenopausal women) failed to lower urinary pH below 5.5 despite the induction of systemic metabolic acidosis. In these patients, therefore, the diagnosis of incomplete distal renal tubular acidosis was made (RTA I). Patients with incomplete RTA I had significantly lower spontaneous plasma pH (7.38 +/- 0.0081 vs 7.41 +/- 0.004, mean +/- SEM, p = 0.002), a lower serum bicarbonate concentration (21.9 +/- 0.49 mmol/l vs 23.1 +/- 0.24 mmol/l, p = 0.034), a lower base excess (-2.33 +/- 0.42 mmol/l vs -0.55 +/- 0.21 mmol/l, p = 0.001) and lower Z-scores in bone densitometry (-2.18 +/- 0.27 vs -1.40 +/- 0.15, p = 0.028) than patients with normal renal acidification. In conclusion, a high prevalence of incomplete RTA I (in 44% of the male patients, 20% of the premenopausal female patients and 6% of all female patients) was found in patients with osteoporosis who, without testing, would have been diagnosed as having 'primary' osteoporosis. The mild metabolic acidosis observed in these patients may have contributed to loss of bone mass by a compensatory mobilization of alkali and calcium from bone. Because of possible therapeutic consequences (e.g., administration of alkali salts and high doses of vitamin D) we propose that measurements of urinary pH and, if necessary, ammonium chloride testing should be included in the diagnostic investigation especially of male and of premenopausal female patients with osteoporosis. Since referral bias, although unlikely, cannot be excluded in our study, the prevalence of RTA I in unselected patients with osteoporosis needs to be determined at primary screening institutions. PMID- 10692984 TI - Treatment of postmenopausal women with osteoporosis or low bone density with raloxifene. Raloxifene Study Group. AB - Raloxifene, a selective estrogen receptor modulator (SERM), has been shown to improved bone mineral density (BMD) and serum lipid profiles in healthy postmenopausal women. The objective of this study was to examine the effects of raloxifene on BMD, biochemical markers of bone metabolism and serum lipids in postmenopausal women with low bone density or osteoporosis. This Phase II, multicenter, 24-month, double-masked study assessed the efficacy and safety of raloxifene in 129 postmenopausal women (mean age +/- SD: 60.2 +/- 6.7 years) with osteoporosis or low bone density (baseline mean lumbar spine BMD T-score: -2.8). Women were randomly assigned to one of three treatment groups: placebo, 60 mg/day raloxifene-HCl (RLX 60) or 150 mg/day raloxifene-HCL (RLX 150) and concomitantly received 1000 mg/day calcium and 300 U/day vitamin D3. At 24 months, BMD was significantly increased in the lumbar spine (+3.2%), femoral neck (+2.1%), trochanter (+2.7%) and total hip (+1.6%) in the RLX 60 group compared with the placebo group (p < 0.05). The RLX 150 group had increases in BMD similar to those observed with RLX 60. A greater percentage of raloxifene-treated patients, compared with those receiving placebo, had increased BMD (p < 0.05). Serum bone specific alkaline phosphatase activity, serum osteocalcin, and urinary type I collagen:creatinine ratio were significantly decreased in the RLX-treated groups, compared with the placebo group (p < 0.01). RLX 60 treatment significantly decreased serum levels of triglycerides, and total- and LDL-cholesterol levels (p < 0.01). The rates of patient discontinuation and adverse events were not significantly different among groups. In this study, raloxifene increased bone density, decreased bone turnover, and improved the serum lipid profile with minimal adverse events, and may be a safe and effective treatment for postmenopausal women with osteoporosis or low bone density. PMID- 10692985 TI - An early-life femoral shaft fracture and bone mineral density at adulthood. AB - High peak bone mass and density in early adulthood is an important protective factor against osteoporotic fractures in later life, but it is not known whether injuries to growing bones adversely affect the attainment of peak bone mass and density. The purpose of this study was therefore to examine with dual-energy X ray absorptiometry the areal bone mineral density (BMD) of the injured and uninjured extremity (the femoral neck, trochanter area of the femur, distal femur, patella, proximal tibia and distal tibia), lumbar spine and distal radius of young adults with a history of an early-life femoral shaft fracture and to find out whether the fracture had affected the attainment of peak bone density of these patients. Thus, the BMD and clinical status of 41 patients (28 men, 13 women) who had sustained a femoral shaft fracture in childhood or adolescence (between 7 and 15 years of age, average 13 years before the study) were examined. The fracture had led to a statistically significant difference in BMD between the injured and uninjured side distal to the fracture site (men/women: distal femur, 3.7%/-3.9%; patella, -3.1%/-5.9%; proximal tibia, -2.0%/-4.6%; distal tibia, 3.4%/-5.2%), whereas the proximal femur did not show such differences. The male patients' spinal BMD was significantly lower (-7.9%) than that in their age-, height- and weight-matched healthy controls. The female patients' spinal BMD tended to be fairly comparable (-1.6%) to that of the controls (NS). In summary, this study indicates that early-life femoral shaft fracture results in a moderate (-2% to -6%) long-term side-to-side BMD difference distal to the fracture site. Patients' spinal BMD values also tend to be lower than that of controls. Thus, a femoral shaft fracture sustained in childhood or adolescence seems to disturb somewhat the attainment of peak bone density, the important predictor of osteoporotic fractures in later life. PMID- 10692986 TI - Toxicity of nitrogen oxides and related oxidants on mycobacteria: M. tuberculosis is resistant to peroxynitrite anion. AB - OBJECTIVE: To test the toxicity of reactive nitrogen intermediates (RNI), including authentic nitric oxide (NO), nitrogen dioxide (NO2), and peroxynitrite anion (ONOO-), a potent oxidant derived from NO and superoxide anion, on various mycobacterial strains including M. tuberculosis. DESIGN: Relatively avirulent mycobacteria including M. smegmatis and BCG, as well as the pathogenic M. Bovis Ravenel and M. tuberculosis Erdman and the clinical isolate M160 (also known as the C strain) were tested for their susceptibility to the toxic effects of NO, NO2, and ONOO-, Deaerated, NO-saturated solutions as well as an anaerobic in vitro system in which mycobacteria can be exposed to desired concentrations of authentic NO or NO2, were employed in these studies. An in vitro ONOO- killing assay was used to examine the adverse effects of this NO-derived oxidant on the various strains of mycobacteria. RESULTS: Both NO and NO2 exhibit antimycobacterial activity, with the former being more potent. Results obtained using ONOO- killing assay revealed that while avirulent mycobacteria including BCG and M. smegmatis are susceptible to this NO-derived oxidant, the virulent Erdman strain of M. tuberculosis and M. bovis, as well as the clinical tuberculous isolate M160, are remarkably resistant. CONCLUSION: These results suggest that the interactions between RNI and various species of mycobacteria could be highly specific. And since activated macrophages produce peroxynitrite, the significance of the ONOO- resistance of M. tuberculosis strains in relation to intracellular survival deserves further investigation. PMID- 10692987 TI - Influence of HLA-DR antigens on lymphocyte response to Mycobacterium tuberculosis culture filtrate antigens and mitogens in pulmonary tuberculosis. AB - SETTING: Influence of HLA-DR antigens and lymphocyte responses in pulmonary TB patients. OBJECTIVE: To elucidate the role of HLA-DR genes/gene products on lymphocyte responses to Mycobacterium tuberculosis antigens and mitogens, the present study was carried out in pulmonary tuberculosis during active and cured stage of the disease. DESIGN: Serological determination of HLA-DR antigens was carried out in 50 active TB patients, 44 cured TB patients and 58 normal healthy control subjects. The influence of HLA-DR antigens on peripheral blood lymphocyte responses to M. tuberculosis culture filtrate antigens and mitogens such as phytohaemagglutinin (PHA) and concanavalin-A (Con-A) was studied in the patients as well as normal healthy control subjects. RESULTS: Of all the DR antigens studied, patients (active TB and cured TB) with DR2 antigen showed an increased lymphocyte response (stimulation index) to a higher dose of antigenic (10 micrograms/ml) stimulation. A significantly lower lymphocyte response to antigen and mitogens was seen in HLA-DR3 positive normal healthy subjects than non-DR3 (DR3 negative) subjects. CONCLUSION: The present study suggests that HLA-DR genes/gene products may be playing an immunoregulatory role in eliciting an immune response against M. tuberculosis antigens and mitogens induced lymphocyte response in pulmonary TB patients and normal healthy subjects. PMID- 10692988 TI - Effects of modulating TGF-beta 1 on immune responses to mycobacterial infection in guinea pigs. AB - SETTING: TGF-beta 1 has been implicated as an important mediator of immuno suppression in clinical tuberculosis. OBJECTIVE: The objective was to determine the role of TGF-beta 1 in experimental pulmonary tuberculosis in the guinea pig. DESIGN: Groups of guinea pigs, maintained on either a low protein (LP) diet or an isocaloric high protein (HP) diet, were challenged via the respiratory route with virulent Mycobacterium tuberculosis H37Rv. Ten days post-infection, guinea pigs were given daily intraperitoneal injections of recombinant human TGF-beta 1 (rhTGF-beta 1 tau for 10 consecutive days). Following the treatment, guinea pigs were euthanized, and PPD-induced proliferation of peripheral blood mononuclear cells (PBMCs) was assessed and disease resistance measured by recovery of mycobacteria from the lungs and spleens. In a second set of experiments, groups of HP and LP guinea pigs were vaccinated with attenuated M. tuberculosis H37Ra. Six weeks later, the effects of rhTGF-beta 1 on lymphoproliferation and cytokine production were determined. RESULTS: Protein deficiency significantly impaired host anti-tuberculosis resistance, as expected. Treatment with rhTGF-beta 1 significantly increased mycobacterial loads in the tissues of guinea pigs and decreased the PPD-induced proliferation of PBMCs from both LP and HP guinea pigs. PPD-driven lymphoproliferation, TNF-alpha and IFN production were significantly suppressed in vaccinated, protein-deficient guinea pigs, and rhTGF-beta 1 further inhibited lymphoproliferation and cytokine production. CONCLUSION: Both in vivo and in vitro results indicate that TGF-beta 1 exerts immunosuppressive activity and exacerbates the progression of experimental pulmonary tuberculosis in both normally nourished and protein-deficient guinea pigs. PMID- 10692989 TI - Increased absolute number but not proportion of gamma/delta T-lymphocytes in the bronchoalveolar lavage fluid of patients with active pulmonary tuberculosis. AB - SETTING: The proportions and absolute cell count of gamma/delta T-lymphocytes in the peripheral blood of patients with pulmonary tuberculosis (PTB) remains controversial. Since PTB is an infections airway disease, bronchoalveolar T lymphocytes should be a better indicator of local immune T-cell reaction after TB infection than peripheral blood T-lymphocytes. OBJECTIVE: To quantitate the absolute cell count and proportions of gamma/delta T-lymphocytes in the bronchoalveolar lavage fluid (BALF) of patients with active PTB. DESIGN: Bronchoalveolar lavage (BAL) and analysis of lymphocytes in the BALF was performed in 25 patients with active PTB and 16 normal controls. All of the patients were negative for HIV infection and none was immunocompromised. BALF and blood were prepared for cell differential count and flow cytometry analysis using monoclonal antibodies CD3, CD4, CD8, CD25, HLA-DR and gamma/delta as well as alpha/beta T-lymphocyte receptors. RESULTS: The number of cells per volume of recovered BALF was significantly higher in the patients with active PTB than in normal controls. BALF from active PTB patients also showed increased percentage of lymphocytes and neutrophils. The absolute number of total lymphocytes, CD3+ lymphocytes and CD3+ gamma/delta T-lymphocytes were significantly higher in the BALF, but not in the blood, of patients with TB, however, the proportions of CD3+ gamma/delta T-lymphocytes in BALF of patients with TB was comparable to that of normal controls. gamma/delta T-lymphocytes in the BALF rarely expressed CD4, CD25, and HLA-DR in both groups. CONCLUSION: These results suggest that gamma/delta T-lymphocytes are not the major subpopulation of CD3+ lymphocytes in the BALF that react to mycobacterial infection in the patients with clinically established active TB. PMID- 10692990 TI - Association of functional mutant homozygotes of the mannose binding protein gene with susceptibility to pulmonary tuberculosis in India. AB - SETTING: Mannose binding protein gene polymorphism in pulmonary tuberculosis in India. OBJECTIVE: To find out whether non-HLA genes such as mannose binding protein (MBP) genes are associated in the susceptibility to pulmonary tuberculosis. DESIGN: Genotyping of MBP 52, 54 and 57 wild and mutant alleles was carried out in HLA-DR typed pulmonary tuberculosis patients (n = 202) and control subjects (n = 109). Since HLA-DR2 is associated with pulmonary-TB, the interaction of MBP genes on -DR2 and non-DR2 genes on the susceptibility was also studied. RESULTS: A significantly increased genotype frequency of MBP functional mutant homozygotes (including 52, 54 and 57) was seen in pulmonary tuberculosis (PTB) patients (10.9%) than in control subjects (1.8%; P = 0.008; odds ratio: 6.5). Analysis of interaction of MBP genes and HLA-DR2 on the susceptibility to PTB revealed that these genes are associated with PTB independent of each other. CONCLUSION: The present study shows that functional mutants of MBP are associated with PTB. Apart from HLA-DR2 association, association of non-HLA genes in the susceptibility to PTB is evident. This suggests that multigenetic factors (candidate genes) may be involved in the susceptibility/resistance to PTB. PMID- 10692991 TI - Diagnostic efficacy of polymerase chain reaction in granulomatous uveitis. AB - SETTING: The granulomatous uveitis, multifocal choroiditis and periphlebitis have been suspected to be of tubercular origin but no definitive reports about detection of etiological agents have been documented in the literature. Conventional bacteriological methods are not generally helpful in diagnosing ocular tuberculosis due to difficulty with potential morbidity associated with obtaining the biopsy material from the eye. Thus, the diagnosis of ocular tuberculosis is most often presumptive. OBJECTIVE: We evaluated the role of polymerase chain reaction (PCR) for detection of Mycobacterium tuberculosis in the aqueous humor samples obtained from eyes with active uveitis. METHODS: Aqueous samples from 53 patients having cellular reaction in the anterior chamber along with any one or more of the following: 1) active vasculitis; 2) anterior vitreous cells; 3) snowball opacities; 4) snow banking in the pars plana; 5) retinochoroiditis were withdrawn by anterior chamber paracentesis and subjected to PCR. Seventeen samples from patients with definite clinical diagnoses other than tuberculosis formed a disease control group. Fifteen aqueous samples obtained from healthy subjects undergoing routine cataract surgery served as healthy controls. PCR was performed using primers capable of amplifying a 150 b.p. segment from a conserved repetitive sequence in the genome of M. tuberculosis. RESULTS: Twenty out of the 53 samples (37.7%) in the study group were positive where as only one sample out of 17 in the disease control group (5.7%) showed a weakly positive band. No sample from the healthy control group showed a positive PCR. CONCLUSION: Our study shows that PCR can be effectively used for the diagnosis of intraocular tuberculosis in the presence of uveitis. PMID- 10692992 TI - Hypodense alveolar macrophages in patients with diabetes mellitus and active pulmonary tuberculosis. AB - SETTING: Alveolar macrophages (AM), a heterogeneous cell population, play a critical role in eliminating mycobacterial infections in collaboration with lymphocytes. Patients with diabetes mellitus (DM) show increased susceptibility to pulmonary tuberculosis (TB) infection. It is still uncertain whether there is a defect in T cell or AM activation in patients with DM against TB infection. OBJECTIVE: To study the difference in activation status of AM and T cells between patients with TB + DM and TB alone. METHOD: The heterogeneity of AM from 14 patients with TB + DM, 9 with TB alone, 10 normal subjects and 8 DM alone patients, was studied using Percoll density fractionation. The intracellular H2O2 production of AM before and after stimulation with phorbol myristate acetate (PMA) or F-Met-Leu-Phen (FMLP) was assayed by loading cells with 2',7' dichlorofluorescin (DCFH) and analyzed by flow cytometry. Lymphocytes subsets (CD3, CD4, CD8) and their activation status (CD25) in bronchoalveolar lavage were also measured. RESULTS: The proportion of the least dense AM (< 1,030 g/ml) and the magnitude of DCFH oxidation of AM was higher in TB patients than in normal subjects, regardless of DM. Patients with TB + DM had a significantly lower proportion of the least density AM fraction than TB alone patients, regardless of disease extent. Among TB patients, the proportion of the least dense AM was inversely correlated with the bacterial load on sputum and the disease extent on chest radiograph. Stimulation of AM with PMA or FMLP induced an increase in the hypodense AM subpopulations and enhanced intracellular H2O2 generation in patients with TB + DM and to a similar extent in normal subjects, but not in patients with TB alone. There was no significant difference in CD3 numbers, CD4/CD8 ratio, and CD25+ cells between patients with TB alone and TB + DM. The activation status of AM or T lymphocytes from DM alone patients was not significantly different from those from normal subjects. CONCLUSION: Hypodense subpopulations of AM increase in active TB patients and are related to the disease severity as well as activation status of AM. AM in TB patients complicated with DM was less activated, and may be contributory to the susceptibility to mycobacterial infection. PMID- 10692993 TI - Development of the Mycobacterium bovis BCG vaccine: review of the historical and biochemical evidence for a genealogical tree. AB - The original Mycobacterium bovis Bacillus Calmette Guerin vaccine strain has developed into several different substrains which have been used for production of BCG vaccines throughout the world since 1921. Based on the latest genetic and antigenic knowledge, as well as the early literature reports on BCG vaccination, we are able to fit the different pieces of the BCG puzzle together and outline the origin of the different substrains of M. bovis BCG. The BCG vaccine substrains analysed demonstrate two distinct patterns, with an abrupt change consisting of a loss of several genes and altered biochemical characteristics in strains originating from Institut Pasteur after 1927. Further evidence from the literature is provided that a change occurred in virulence of the BCG parent strain at Institut Pasteur in the late 1920s. Based on this information a genealogical tree is proposed and discussed. PMID- 10692994 TI - Immunogenicity and protective efficacy of DNA vaccines encoding secreted and non secreted forms of Mycobacterium tuberculosis Ag85A. AB - OBJECTIVE: To determine the efficacy of Ag85A-DNA against challenge with a highly virulent human clinical isolate of Mycobacterium tuberculosis (CSU37) and to compare the potencies of two types of Ag85A-DNA vaccines; those expressing secreted and non-secreted forms of the protein. DESIGN: Ag85A-DNA vaccinated mice were challenged with a highly virulent clinical isolate of M. tuberculosis (CSU37) in order to compare the efficacy of these vaccines. In vitro studies were also performed. RESULTS: Enhanced humoral and cellular responses were induced in mice vaccinated with the secreted Ag85A-DNA compared to the non-secreted Ag85A DNA. In addition, secreted Ag85A-DNA conferred protective immunity against infection with M. tuberculosis (CSU37). CONCLUSIONS: DNA vaccines encoding M. tuberculosis Ag85A have been shown to induce potent humoral and cellular immune responses leading to protection from M. tuberculosis (Erdman) challenge in mouse models. In this study we demonstrate that Ag85A can confer protection in a rigorous challenge model using a highly virulent human clinical isolate of M. tuberculosis (CSU37). This challenge model appears able to discriminate between DNA vaccines of differing potencies, as the more immunogenic DNA construct encoding a secreted form of Ag85A was protective, whereas the less immunogenic DNA construct encoding a non-secreted form of Ag85A was not. PMID- 10692995 TI - Mechanisms of vaccine-induced resistance in a guinea pig model of pulmonary tuberculosis. PMID- 10692996 TI - The molecular basis of isoniazid resistance in Mycobacterium tuberculosis. PMID- 10692997 TI - Rapid kinetic spectrophotometric determination of phosalone (Zolone) in a commercial formulation. AB - A kinetic study of the degradation of phosalone in an alkaline medium was undertaken by using a pneumatic stopped-flow system. A rapid semiautomatic method is proposed for determining phosalone. Linear calibration graphs up to 8.0 x 10( 5) M (detection limit = 1.40 x 10(-6) M) were obtained, with a measurement period of only 3.5 s per sample and a relative standard deviation of 1.4%. Several pesticides were assayed as interference species, and several did not interfere even at a 6:1, M:M foreign species/phosalone ratio. A strong interference (ratio < 1) was generated by azinphos-methyl and carbaryl. The proposed method was applied to the analysis of a commercial formulation, and the results were validated by comparison with those for a chromatographic method. PMID- 10692998 TI - Determination of histidine and related compounds in rumen fluid by liquid chromatography. AB - A liquid chromatographic procedure was developed for quantitative determination of histidine (His), histidinol (HDL), histamine (HTM), urocanic acid (URA), imidazolepyruvic acid (ImPA), imidazoleacetic acid (ImAA), and imidazolelactic acid (ImLA) in rumen fluid. The method is based on direct injection analysis by UV absorbance detection at 220 nm. The separation was performed under 2 different chromatographic conditions on a LiChrospher 100 NH2 column. In the first chromatographic system, the mobile phase used for isocratic elution was 67 mM potassium phosphate buffer (monobasic and dibasic) pH 6.45-90% acetonitrile in water (21 + 79); in the second system, an acetonitrile gradient in 63 mM potassium phosphate buffer (monobasic) pH 3.0, obtained by addition of 60 mM phosphoric acid, was used. Analyses of both systems were completed within 32 and 25 min, respectively. The limits of detection of these compounds were (microM): His, 2.8; HDL, 3.7; HTM, 4.0; URA, 0.75; ImPA, 4.7; ImAA, 1.2; and ImLA, 1.3. Recovery of these compounds added to rumen fluid was 97.4-103.0% within a 1-day study and 95.4-99.0% on different day studies. Detectable levels of His were found in the deproteinized rumen fluid of goats, with average concentrations of 16.10, 10.43, 11.14, and 13.62 microM in the rumen fluid collected before the morning feeding and 2, 4, and 6 h after feeding, respectively. HDL, HTM, URA, ImPA, ImAA, and ImLA were not detected in the rumen fluid before and after feeding. Trp, Phe, and Tyr were also identified in the rumen fluid, with average concentrations of 8.25, 29.04, and 12.6 microM, respectively, before the morning feeding. PMID- 10692999 TI - Rapid determination of nosiheptide in meat and egg by liquid chromatography with fluorescence detection. AB - A procedure was developed to determine nosiheptide residues in marketed meat and egg. Acetonitrile was used for the extraction, and the extract was partitioned with hexane to remove fat. The lower layer was reconstructed and quantitated by liquid chromatography using fluorescence detection at 357 nm excitation and 500 nm emission. The mobile phase consisted of 0.025% phosphoric acid-acetonitrile (50 + 50, v/v). Recoveries of nosiheptide from fortified samples ranged from 91.3 to 95.2% for swine muscle, 88.6 to 92.7% for chicken muscle, and 86.3 to 86.8% for egg. The method was used to monitor swine and chicken muscle and egg (20 samples each) in the market. Nosiheptide was not determined in all 60 samples. PMID- 10693000 TI - Determination of amoxicillin residues in animal tissues by solid-phase extraction and liquid chromatography with fluorescence detection. AB - Trace levels of amoxicillin residues were determined in animal tissues by liquid chromatography (LC) with fluorescence detection. An improved solid-phase extraction (SPE) procedure requiring less flammable solvent (diethyl ether) was developed for sample preparation. Muscle samples of beef, pork, chicken, and tilapia were extracted with a phosphate buffer followed by the modified SPE procedure for cleanup and concentration prior to the LC-fluorescence analysis. Average recoveries of fortified amoxicillin at 5, 10, and 20 micrograms/kg ranged from 83.9 to 85.8% in beef, 86.1 to 88.1% in pork, 81.7 to 82.9% in chicken, and 92.5 to 95.4% in tilapia. Relative standard deviations were < 4%. PMID- 10693001 TI - Simultaneous determination of residues of chloramphenicol, florfenicol, florfenicol amine, and thiamphenicol in shrimp tissue by gas chromatography with electron capture detection. AB - A gas chromatographic (GC) method is presented for determining residues of chloramphenicol (CAP), florfenicol (FF), florfenicol amine (FFa), and thiamphenicol (TAP) in shrimp tissues, with meta-nitrochloramphenicol (mCAP) as the internal standard. The composited shrimp is extracted with basic ethyl acetate, followed by an acetonitrile-basic ethyl acetate mixture. This extract is centrifuged, filtered, evaporated, and reconstituted in water; the reconstituted extract is acidified, defatted with hexane, and passed through a propylsulfonic acid (PRS) and C18 solid-phase extraction (SPE) system. The C18 SPE column is eluted with methanol, and the PRS SPE column is eluted with basic MeOH plus counter ion. The combined eluates are evaporated, reconstituted in acetonitrile, and derivatized with Sylon BFT. After derivatization, the addition of toluene directly to the sample, followed by the addition of basic water, quenches the derivatization process. After centrifugation, the organic layer is carefully removed, and the analytes are determined by GC with electron capture detection. Shrimp tissues were fortified with fenicols (i.e., CAP, FF, FFa, and TAP) at 5, 10, 20, 40, and 80 ng/mL. Overall recoveries were 88, 101, 91, and 84% with overall interassay (between-day) variabilities (i.e., relative standard deviations) of 5.3, 9.4, 12.8, and 7.4% for CAP, FF, FFa, and TAP, respectively. The method detection limits were calculated as 0.7, 1.4, 2.4, and 1.3 ng/g (ppb) for CAP, FF, FFa, and TAP, respectively, based on a 10 g sample. The quantitation limit as determined empirically by this method is the lower limit of the standard curve, which is about 5 ng/g (ppb) for each analyte. PMID- 10693002 TI - Simultaneous determination of eprinomectin, moxidectin, abamectin, doramectin, and ivermectin in beef liver by LC with fluorescence detection. AB - Eprinomectin, moxidectin, abamectin, doramectin, and ivermectin are drugs used to control parasitic infections in both meat-producing and nonmeat-producing animals. A number of analytical methods are available to analyze these anthelmintic drugs individually. A multiresidue screening method was developed for these drugs; however, the initial attempt to derivatize eprinomectin following the method published by Merck scientists was unsuccessful because the eprinomectin derivatization reaction was temperature- and time-dependent. The optimum time and temperature for the completion of eprinomectin derivatization were 90 min and 65 degrees C, respectively, without appreciable effect on the remaining 4 drugs. Beef liver samples were fortified with 0, 25, 50, and 100 ppb mixed standards of eprinomectin, moxidectin, abamectin, doramectin, and ivermectin. Each set of 4 levels of recoveries was repeated 10 times with all 5 compounds. The average of 10 recoveries of 5 compounds at all 4 levels of fortification was > 70%; the coefficient of variation was < 20%. PMID- 10693003 TI - Confirmation of eprinomectin, moxidectin, abamectin, doramectin, and ivermectin in beef liver by liquid chromatography/positive ion atmospheric pressure chemical ionization mass spectrometry. AB - A liquid chromatographic (LC) multiresidue screening procedure was developed for determination of eprinomectin, moxidectin, abamectin, doramectin, and ivermectin in beef liver at 0, 25, 50, and 100 ppb levels. A procedure using low resolution LC/atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) was developed with further purification steps added to the quantitative LC method to confirm residues. Acetonitrile extracts of liver, prior to derivatization for LC analysis, were further purified by using a C8 solid-phase extraction cartridge and an alumina-B cartridge. The purified extract was analyzed by injection into an LC/positive ion APCI MS. Identity of the compound was confirmed by comparison of its retention time and relative intensity data with those of a standard or recovery from a fortified control liver sample. Anthelmintic drugs in acetonitrile extracts of liver containing eprinomectin, moxidectin, abamectin, doramectin, and ivermectin at 25 ppb, the lowest level of fortification used in the LC determinative method, were successfully confirmed. PMID- 10693004 TI - Comparative photodegradation study of atrazine and desethylatrazine in water samples containing titanium dioxide/hydrogen peroxide and ferric chloride/hydrogen peroxide. AB - Results are reported for a comparative photodegradation study of atrazine and desethylatrazine in water using TiO2/H2O2, FeCl3/H2O2, and photolysis. Deionized water and ground water spiked with atrazine or desethylatrazine at 36 micrograms/L were irradiated by using a xenon arc lamp and/or sunlight. After irradiation, the water samples containing the spiked pesticides were preconcentrated by using C18 solid-phase extraction disks and analyzed by gas chromatography with nitrogen-phosphorus and mass spectrometric detection. A relative percentage of 7% desethylatrazine was detected in samples removed after 20 and 4 min of sensitized photodegradation with TiO2 and Fe3+, respectively. Atrazine and desethylatrazine did not degrade when solar irradiation (in winter) and deionized water were used. Atrazine degraded faster than desethylatrazine when a xenon arc lamp or sunlight plus FeCl3 was used, with half-lives varying from 5 to 11 min and from 19 to 26 min, respectively. In other photodegradation experiments, the degradation of atrazine was slightly higher than that of desethylatrazine. This study shows that desethylatrazine has slightly higher stability than atrazine in environmental water samples; this stability accounts for the frequent detection of desethylatrazine together with atrazine in natural waters. PMID- 10693005 TI - New monitoring system for ninety pesticides and related compounds in river water by solid-phase extraction with determination by gas chromatography/mass spectrometry. AB - A new monitoring system was established for the determination of 90 pesticides and 10 pesticide degradation products in river water. The pesticides consisted of 18 fungicides, 30 insecticides, and 42 herbicides. The pesticides were extracted with a solid-phase, styrene-divinylbenzene copolymer, eluted with acetone, hexane, and ethyl acetate, and determined by gas chromatography/mass spectrometry. Overall recoveries ranged from 72 to 118%. The limits of detection were 0.01-0.1 microgram/L. This system determines most of the pesticides used in Japan and was successfully applied to practical monitoring of water polluted with pesticides and related compounds. PMID- 10693006 TI - Detection of pork in heat-processed meat products by monoclonal antibody-based ELISA. AB - An enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody to a porcine thermal-stable muscle protein was developed for detection of pork in cooked meat products. The assay specifically detects porcine skeletal muscle, but not cardiac muscle, smooth muscle, blood, and nonmuscle organs. No cross reactivity was observed with common food proteins. Validity of the assay was evaluated with laboratory formulated and commercial meat samples. The detection limit was determined as 0.5% (w/w) pork in heterologous meat mixtures. Overall, intra- and inter-assay coefficients of variation were 5.8 and 7.9%, respectively. The accuracy in analyzing market samples was 100% as verified by product labeling and confirmed by a commercial polycolonal antibody test kit. PMID- 10693007 TI - Rapid determination of Listeria monocytogenes by automated enzyme-linked immunoassay and nonradioactive DNA probe. AB - A rapid and reliable analytical method was developed to detect and confirm the presence of Listeria monocytogenes in raw and partially processed foods. Forty nine food samples (25 mixed cut vegetable salad, 12 smoked salmon, and 12 sterile smoked salmon) were individually inoculated with high levels [10-100 colony forming units (cfu)/25 g sample] and low levels (1-10 cfu/25 g sample) of L. monocytogenes, and were screened using the Vitek Immuno Diagnostic Assay (VIDAS) Listeria monocytogenes (VIDAS LMO)]. Positive test results were confirmed as L. monocytogenes by nonradioactive DNA probe. All samples inoculated with high levels of L. monocytogenes were detected by VIDAS and 96% were confirmed as L. monocytogenes by DNA probe. VIDAS LMO detected 89% of samples inoculated with low levels of L. monocytogenes, and 87% of these were confirmed as positive by DNA probe. In addition, 12 other samples (4 from each of mixed cut vegetable salad, smoked salmon, and sterile smoked salmon) were inoculated with high levels of L. ivanovii, L. seeligeri, L. welshimeri, L. innocua, L. grayi, and L. murrayi. Samples were assayed by the same protocol and all gave negative results. Compared with the cultural method, the VIDAS LMO nonradioactive DNA probe combination is highly specific, discriminates between L. monocytogenes and all other Listeria species, and reduces analytical time. PMID- 10693008 TI - Determination of histamine in tomatoes by capillary electrophoresis. AB - An improved capillary electrophoresis assay for histamine in crude extracts was developed and used to determine histamine levels in a selection of tomato fruits and pastes. Performance in terms of reproducibility and sensitivity was optimized by use of a high sensitivity detector flow cell, sodium hydroxide rinses, and a voltage gradient. The method was linear down to 0.2 microgram/mL (signal-to-noise ratio = 4:1), which was below the endogenous level in all samples. PMID- 10693009 TI - Determination of aflatoxins in grains and raw peanuts by a rapid procedure with fluorometric analysis. AB - A rapid, quantitative, inexpensive, and efficient method was developed to determine aflatoxins in corn, corn meal, popcorn, rice, wheat, cottonseed, and peanuts. Samples are ground and extracted with methanol-water (80 + 20). A portion of the extract is cleaned up by passage through a solid-phase separatory column, 500 microL purified extract is derivatized with a bromine reagent, and fluorescence of the solution is immediately quantified with a calibrated fluorometer containing a broad wavelength pulsed xenon light source. This method can quantify aflatoxin from 5 to 5000 ppb without dilution and was linear when applied to samples of noncontaminated corn spiked at 0 to 5000 micrograms aflatoxin B1/g. Correlation coefficients of the method with LC for multiple analyses for corn (n = 34), cottonseed (n = 32), and peanuts (n = 11) were 0.999, 0.995, and 0.980, respectively. Individual analyses may be conducted in less than 5 min, and grouping of samples is unnecessary. The sensitivity of the method for corn is 5 ppb and the fluorometer, under the operating conditions, has a limit of detection of 0.6 ng aflatoxin B1. PMID- 10693010 TI - Determination of fumonisins B1 and B2 in various maize products by a combined SAX + C18 column and immunoaffinity column. AB - Fumonisins B1 and B2 were determined in 42 samples of different maize products from the Swedish market by 2 different methods based on cleanup steps using an immunoaffinity column and a combination of SAX + C18 columns, respectively. A simple "precipitation step" was included before the samples were added to the main column(s), giving less column clogging, fewer interfering peaks, and better recoveries for the different sample matrixes. Recovery, repeatability, and results from the survey showed comparable results with the methods. The limit of detection for both methods was 5 micrograms/kg for fumonisin B1 and 10 micrograms/kg for fumonisin B2. All 7 maize chips analyzed and 6 of 8 popcorn samples contained fumonisins (B1 + B2) with averages of 180 and 115 micrograms/kg, respectively. All other samples except a maize flour sample contained little or no fumonisins. PMID- 10693011 TI - Capillary gas chromatography/mass spectrometry with chemical ionization and negative ion detection for confirmation of identity of patulin in apple juice. AB - Gas chromatography/mass spectrometry (GC/MS) with negative ion chemical ionization permits detection of underivatized patulin in apple juice extracts while minimizing co-extractive responses. The technique has been used with a variety of capillary columns in quadrupole, ion trap, and magnetic sector GC/MS instruments to confirm presumptive findings of patulin in apple juice at concentrations ranging from 68 to 3700 micrograms/L. The demonstrated ability to use any of these 3 mass spectrometers and several capillary columns to confirm the identity of patulin are significant strengths of the technique. PMID- 10693012 TI - Determination of low levels of mite and insect contaminants in food and feedstuffs by a modified flotation method. AB - Extraneous material was separated from feed and food products by a modified technique in which kerosene is used in a specially designed flotation flask. This technique, although effective for analyzing feed and foods, presented limitations in the analysis of finely powdered materials. Some procedural modifications and an increased in the capacity of the flotation flask from 500 to 750 mL allowed a larger sample weight (20 g) to be analyzed for mites, insect fragments, and rodent hairs, with considerably reduced residue interference. In trials with a variety of products seeded with known numbers of mites, average recovery was 83%. Recoveries of 89% were obtained from flour samples seeded with insect fragments and rodent hairs. A new process of suspending extracted mites in a mixture of industrial methylated spirit (46%) and glycerol (54%) by volume was used to allow rapid and more precise estimates of mite populations in heavily infested samples. PMID- 10693013 TI - Determination of vitamin K in milk and infant formulas by liquid chromatography: collaborative study. AB - A simple procedure for determination of vitamin K1 was developed for routine compliance monitoring of supplemented infant formula and measurement of endogenous levels in milk and milk powders. Samples are digested with lipase and extracted into hexane; and aliquot is evaporated, reconstituted in methanol, and analyzed by reversed-phase LC. Post-column zinc reduction of phylloquinone facilitates detection by fluorescence. The procedure was subjected to an AOAC collaborative study involving 8 materials, each in blind duplicate, across the range of 5-120 micrograms/100 g solids and including NIST 1846 reference material. Thirty-three laboratories returned valid data which were then statistically analyzed for outliers and precision parameters. Mean RSDR (%) was 6.53 (4.33-10.94), with a mean HORRAT value of 0.33 (0.23-0.43) and RSDr:RSDR ratio of 0.74. K1 isomers (cis and trans) were aggregated with conventional C18 columns, but may be selectively estimated with use of the C30 column. PMID- 10693014 TI - Determination of choline in milk and infant formulas by enzymatic analysis: collaborative study. AB - A collaborative study was conducted on a coupled enzymatic-spectrophotometric method for the determination of choline in infant formula and milk powders. Twenty-nine laboratories participated in the analysis of 8 blind duplicates over the range of 45-175 micrograms/100 g sample. After the combined acid hydrolysis phospholipase release of choline, incubation with choline oxidase in the presence of peroxidase and phenol generates a quinoneimine chromophore with 4 aminoantipyrine. Following raw data screening, overall mean RSDR was estimated at 5.14% (range, 4.26-6.34%) with a HORRAT value of 0.91 (range, 0.76-1.00) and an overall mean RSDr:RSDR value of 0.53 (range, 0.42-0.74). The method was also compared with alternative independent analytical techniques for several of the collaborative study samples. PMID- 10693015 TI - Determination of egg proteins in food products by enzyme immunoassay. AB - An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of egg proteins in foods. The polyclonal antibodies developed were specific to whole egg proteins and did not cross-react with any of the 38 nuts, legumes, or other common food ingredients tested. The concentrations of egg proteins that will inhibit 50% of antibody-antigen binding, IC50, were 3-7 ng/mL, and the linear range was 0.5-62.5 ng/mL. The detection limit was 0.2 ppm for various foods. Recoveries ranged from 67 to 96%. The intra- and inter-assay coefficients of variation in this procedure were 10-13% for ice cream spiked at 0.8 and 1.6 ppm. The ELISA has been applied to ice creams, noodles, pasta, and breads. Egg proteins were identified in all declared egg products, and no false positives were found. PMID- 10693016 TI - Recovery and sporicidal resistance of various B. subtilis spore preparations on porcelain penicylinders compared with results from AOAC test methods. AB - Sporicidal test results obtained from carriers inoculated with 4 types of defined Bacillus subtilis spore preparations were compared with the standard AOAC sporicidal test using soil extract nutrient broth (SENB) B. subtilis 19659 spores. Recoveries of spores inoculated on penicylinders from B. subtilis clean spores (washed and suspended in water) and B. subtilis 19659 spores inoculated from culture filtrates according to the AOAC method were compared. Spores were exposed to 6 concentrations (0.5-3.0% w/v) of glutaraldehyde in phosphate buffer (pH 7.5) for 10 h. Concentrations were established by titrimetry and liquid chromatography. Recoveries of surviving spores were determined for 3 types of clean B. subtilis var. niger preparations, one clean B. subtilis 19659 preparation, and the SENB B. subtilis 19659 filtrates. Spore carriers, inoculated by the standard AOAC protocol, resulted in as much as a 2-log number difference in runs 1-12, but not more than 0.5 log number for each clean spore preparation. The SENB spores varied most in resistance to glutaraldehyde, with no growth in recovery media from 3 different batches of 1, 1.5, and 2% glutaraldehyde. Separate batches of SENB preparations of B. subtilis 19659 were resistant and destroyed by 1.0% glutaraldehyde, with 3.98 and 6.0 log numbers of spores on penicylinders, respectively. Clean spore preparations of B. subtilis 19659 on porcelain penicylinders were more resistant to glutaraldehyde than were SENB spores. Nutrient agar/Mg/Ca and nutrient agar/Mg spore preparations of B. subtilis var. niger showed the most uniform resistance to glutaraldehyde. Spores with calcium added showed increased resistance to glutaraldehyde. B. subtilis 19659 spores from the Columbia broth spore preparation were the most resistant and were recovered after exposure to 3.0% glutaraldehyde. PMID- 10693017 TI - Element and radionuclide concentrations in food: FDA Total Diet Study 1991-1996. AB - Food purchased throughout the United States during 1991-1997 under the U.S. Food and Drug Administration's Total Diet Study (TDS) program were analyzed for elements and radionuclides. The program is described with emphasis on food analysis and quality control, including independent interlaboratory exercises. Analytical results are summarized for Cd, Pb, Ni, As, Hg, Se, Cu, Zn, Mn, Fe, Mg, Ca, P, K, and Na and for 137Cs, 131I, 106Ru, and 90Sr. Concentration data are provided to expand the information base used to support assessments of the safety and nutritive value of the U.S. food supply and for their potential use in food composition databases. For selected foods, comparisons were made with past TDS results and with those reported in the literature. An extensive listing of the analytical data is available on the FDA CFSAN Website. PMID- 10693018 TI - Determination of malathion, coumaphos, and fluvalinate residues in honey by gas chromatography with nitrogen-phosphorus or electron capture detectors. AB - A rapid, reliable, and inexpensive extraction method was developed to determine acaricide residues in honey by gas chromatography (GC) with nitrogen-phosphorus (NP) or electron capture (EC) detectors. Because of the high selectivity of the NP detector, no interfering peaks were present and no cleanup was necessary. A simple cleanup step is proposed for the GC-ECD analysis. Recoveries from spiked honey samples ranged from 79 to 94.4%, with coefficients of variation of 0.3 18.5%. The quantitation limit obtained was 0.015 mg/kg for malathion, 0.020 mg/kg for coumaphos, and 0.005 mg/kg for fluvalinate. The method was used to determine the disappearance of malathion and coumaphos residues from honey samples collected from beehives treated with these acaricides. The disappearance of both acaricides was rapid and followed a first-order model for the duration of the experiment. PMID- 10693019 TI - Determination of six metals in galician red wines (in northwestern Spain) by capillary electrophoresis. AB - A simple technique is described for the routine simultaneous capillary electrophoretic determination of 6 cations in wine. Separation was achieved on a fused silica capillary column with a UV-Cat-1, alpha-hydroxysobutyric acid and 18 crown-6-ether buffer at pH 4.5 and indirect UV detection at 214 nm. The content of magnesium, sodium, potassium, calcium, manganese, and lithium was determined. The method is quantitative, with recoveries in the 92-102% range, and linear over more than one order of magnitude. The precision is better than 2.5-3.4%. The method is sensible, with detection limits between 0.01 and 0.06 mg/L. Twenty-five red wines with a Certified Brand of Origin from Galicia (north-western Spain) were analyzed by the proposed method. Various wines showed very similar electrophoretic profiles, but significant quantitative differences were observed. PMID- 10693020 TI - Optimized determination of iron in grape juice, wines, and other alcoholic beverages by atomic absorption spectrometry. AB - This paper describes a study of the different methods of sample preparation for the determination of iron in grape juice, wines, and other alcoholic beverages by atomic absorption spectrometry with electrothermal atomization; results are also reported for the practical application of these methods to the analysis of commercial samples produced in Spain. The methods examined include dealcoholization and dry and wet mineralization treatment using different acids and/or mixtures of them, both with and without heating. The sensitivity, detection limit, accuracy, precision, and selectivity of each method were established. The best results were obtained for wet mineralization with heated acid (HNO3-H2SO4); the results for table wines had an accuracy of 97.5-101.6%, a relative standard deviation of 3.51%, a detection limit of 19.2 micrograms/L, and a determination limit of 32.0 micrograms/L. The method was also sufficiently sensitive and selective. It was applied to the determination of iron in grape juice, different types of wines, and beverages with high alcoholic content, all of which are produced and widely consumed in Spain. The values obtained ranged from 3.394 +/- 2.15 mg/L for the juice, 2.938 +/- 1.47 mg/L for the white wines, 19.470 +/- 5.43 mg/L for the sweet wines, 0.311 +/- 0.07 mg/L for the brandies, and 0.564 +/- 0.12 mg/L for the anisettes. Thus, the method is useful for routine analysis in the quality control of these beverages. PMID- 10693021 TI - Pesticide residues on fruits and vegetables from Ontario, Canada, 1991-1995. AB - For the 5-year period 1991 to 1995, 1536 vegetable and 802 fruit samples were analyzed. The purpose of this study was to determine if pesticides were present on Ontario-produced fruits and vegetables, and if so, to determine if residues violated maximum residue limits (MRLs). Overall, 31.5% of the samples had no detectable pesticide residues, whereas 68.5% contained one or more residues. Most of the residues were present at very low concentrations; 48% of the detections were < 0.1 parts per million (ppm), and 86% were < 1 ppm. However, violations of MRL were observed in only 3.2% of the vegetables samples and 3.1% of the fruit samples. In addition, 4.8% of the samples contained a "technical" violation, that is, there was no specified MRL for the pesticide-commodity combination and the residues exceeded 0.1 ppm. Of the detectable residues, 63% were < 10% of the MRL, whereas 89% were < 50% of the MRL. More fruit samples (91.4%) had a detectable residue, compared with vegetable samples (56.6%). Fruit is often treated close to harvest or post harvest to ensure that wholesome produce reaches the consumer. Forty-six percent of the samples contained 2 or more residues, and 2% of all samples had more than 5 different pesticides detected; fruit samples tended to have more multiple residues. The most frequently found pesticides were captan, the dithiocarbamate fungicides, endosulfan, azinphos-methyl, phosmet, parathion, and iprodione. These pesticides were also used in the greatest quantity for crop production. Overall, the data agree fairly closely with those reported for the U.S. Department of Agriculture Pesticide Data Program because the 2 programs have similar analytical goals and objectives. PMID- 10693022 TI - Quantitative relationship of application rate and pesticide residues in greenhouse tomatoes. AB - The association between application rate of a pesticide and its residue in ripe tomatoes was studied. The average residue level (R) of any pesticide in ripe tomatoes remained in quantitative relation to its dose (D), expressed by the following regression equation: R = 0.24 D (mg/kg), where the numerical factor, 0.24, represents the average residue in mg/kg after application of 1 kg active ingredient per hectare with relative standard deviation of 23%. Quantitative association between these 2 factors enables evaluation of greenhouse tomato growers with respect to their observation of Good Agricultural Practice rules and the Plant Protection Act, obligatory in Poland since 1996, and thus may be a reliable basis for the registration of new agrochemicals. PMID- 10693023 TI - Selective determination of chromium (VI) in powdered milk infant formulas by electrothermal atomization atomic absorption spectrometry after ion exchange. AB - A previously developed method was adapted to the selective determination of hexavalent chromium in powdered milk infant formulas. The species in reconstituted milk was separated on an ion-exchange column, Chromabond NH2, and measured by electrothermal atomization atomic absorption spectrometry. The detection limit was 1.8 micrograms/L, and the linearity range under optimized conditions was 1.8-50.0 micrograms/L. The precision values were 4.1 and 6.5% for the analytical and overall procedures, respectively. The procedure was validated by the method of standard additions (5.0, 10.0, and 25.0 micrograms/L), and the recoveries were all > 93%. The developed method is sensitive, accurate, and precise for determination of Cr(VI) in powdered milks. It was applied to the determination of Cr(VI) in 20 commercial brands, i.e., 7 infant formulas, 5 follow-up milks, and 8 dietetic milks. The values found ranged from < 10 to 75 ng/g. PMID- 10693024 TI - Speciation of iodide, iodine, and iodate in environmental matrixes by inductively coupled plasma atomic emission spectrometry using in situ chemical manipulation. AB - Dissolved iodine, iodide, and iodate are determined in environmental matrixes by in situ chemical manipulation and inductively coupled plasma atomic emission spectrometry (ICPAES). The method uses equipment commonly available to most laboratories involved in environmental inorganic analysis. Total dissolved iodine, iodide, and iodate are determined by ICPAES using iodine vapor generation. Total iodine is determined directly by ICPAES after filtration. Total dissolved iodide (I-) is oxidized in situ to iodine by the addition of sodium nitrite in sulfuric acid in a simplified continuous flow manifold. Iodate is determined by prereduction at the instrument before analysis by the in situ oxidation ICPAES procedure. A standard nebulizer produces the gas-liquid separation of the total iodine, which is then quantified by ICPAES at 206.16 nm. The instrument detection limit for the iodine analysis was 0.04 microgram/mL. Recoveries from seawater, saltwater, and freshwater standard reference materials ranged from 85 to 118% and averaged 98%. For samples containing both iodine and iodide, the total is determined with in situ oxidation, iodine is determined without the oxidizing reagents, and iodine is calculated from the difference. For samples containing all 3 species, pre-reduction is used and the iodine and iodide concentrations are subtracted for quantitation of iodate. The analysis is selective for these 3 species (I-, I2, and IO3). A group of 20-30 samples may be analyzed and quantitated for all 3 individual, commonly occurring iodide species in less than 1 h. The procedure is considerably faster than any other reported techniques. This method is especially well-suited to the analysis of small environmental samples. PMID- 10693025 TI - Improved determination of mercury complex with Thiomicher's ketone by beta correction spectrophotometry. AB - Determination of the mercury complex formed with Thiomicher's ketone (TMK) was improved by beta-correction spectrophotometry in the presence of a nonionic surfactant at pH 5. The complex formed was Hg(TMK)2, and its true molar absorptivity is reported for the first time: epsilon Hg(TMK)2(560) = 1.04 x 10(5) L/mol.cm. In addition, the stability constant of Hg(TMK)2 was equal to 3.64 x 10(10) at an ion strength of 0.01 at 20 degrees C. Results from analyses of wastewater samples showed that the relative standard deviations were < or = 8.3%, and the recoveries of mercury ranged from 90 to 110%. PMID- 10693026 TI - Liquid chromatographic determination of vanillin and related aromatic compounds. AB - This paper describes a reversed-phase liquid chromatographic method for the determination of vanillin, associated natural aromatic compounds and/or synthetic precursors, ethyl vanillin, and coumarin, a commonly encountered adulterant in nonbeverage and beverage alcohol products using a ternary gradient mobile phase. The compounds were separated on a Nova-Pak C18 column by using water, methanol, and tetrahydrofuran as the mobile phase. Measurements were made by using a photodiode array detector at 275 nm. The choice of the mobile phase and the column provides baseline resolution of vanillin and the associated aromatic compounds commonly found in vanilla-flavoring material. Because this method provides low-level detection/quantitation, it is suitable for the characterization of vanilla flavoring materials that are currently added to vanilla flavored beverage alcohol products. PMID- 10693027 TI - Determination of 4-hexylresorcinol in shrimp by liquid chromatography with fluorescence detection. AB - A method was developed to determine 4-hexylresorcinol in shrimp meat. The procedure is based on extraction of test portions with methanol followed by liquid chromatographic analysis of the extracts, using a reversed-phase column and fluorimetric detection (excitation: 280 nm, and emission: 310 nm). The confidence interval of the recovery in working range of 1.5-2.5 mg/kg was 81.6 +/ 0.8%. The relative standard deviation in the working range was 2.1%. Limits of quantitation and detection were 6.59 and 1.98 ng/mL extract, respectively, corresponding to 0.26 and 0.08 mg/kg in shrimp. PMID- 10693028 TI - Assessing ADHD and comorbid disorders in children: the Child Behavior Checklist and the Devereux Scales of Mental Disorders. AB - Evaluated discriminant validity and clinical utility of selected subscales of the Devereux Scales of Mental Disorders (DSMD; Naglieri, LeBuffe, & Pfeiffer, 1994) and the Child Behavior Checklist (CBCL; Achenbach, 1991a) in 228 children referred to a clinic for the evaluation and treatment of attention deficit hyperactivity disorder (ADHD). The DSMD is a multiaxial behavior rating scale that measures symptomatology for a broad range of child psychopathology as described in the Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev. [DSM-R-III] and 4th ed. [DSM-IV]; American Psychiatric Association, 1987, 1994). Discriminant function analyses as well as sensitivity, specificity, and predictive power analyses were computed to evaluate the discriminant validity and clinical utility of selected DSMD and CBCL subscales for assessing ADHD, oppositional defiant disorder (ODD), and anxiety disorders. Results indicated that the DSMD compared very favorably with the CBCL in the ability to discriminate between children with ADHD and those without ADHD and between children with comorbid ODD and anxiety disorders and children who did not meet criteria for these disorders. The DSMD Attention subscale may be somewhat better at ruling in ADHD combined subtype (ADHD-C) and ADHD inattentive subtype (ADHD-I) than the CBCL Attention Problems subscale, but the CBCL Attention Problems subscale may have slightly better utility than the DSMD Attention subscale in ruling out these subtypes. Both the CBCL and DSMD were more useful for ruling out than for ruling in ODD and anxiety disorders. PMID- 10693030 TI - Social functioning and emotional regulation in the attention deficit hyperactivity disorder subtypes. AB - Compared 16 children with attention deficit hyperactivity disorder (ADHD) combined type (ADHD-C), 14 children with ADHD predominantly inattentive type (ADHD-I), and 17 controls on parent and teacher ratings of social status and performance, self-report of social knowledge and performance, and observations of behavior on an emotional regulation task. Analyses revealed distinct patterns of social dysfunction between ADHD subgroups. Children with ADHD-C were rated as showing more aggressive behavior; furthermore, they displayed emotional dysregulation characterized by high intensity and high levels of both positive and negative behavior. In contrast, children with ADHD-I were perceived as displaying social passivity and showed deficits in social knowledge on the self report measure but did not evidence problems in emotional regulation. Regression analyses revealed that social performance, emotional regulation, and, to a lesser degree, social knowledge, were predictive of social status. The application of these findings to understanding the nature of the social deficits in the ADHD subtypes and directions for future research are discussed. PMID- 10693029 TI - Parenting practices and child disruptive behavior problems in early elementary school. Conduct Problems Prevention Research Group. AB - Examined the hypothesis that distinct parenting practices may be associated with type and profile of a child's disruptive behavior problems (e.g., oppositional, aggressive, hyperactive). Parents of 631 behaviorally disruptive children described the extent to which they experienced warm and involved interactions with their children and the extent to which their discipline strategies were inconsistent and punitive and involved spanking and physical aggression. As expected from a developmental perspective, parenting practices that included punitive interactions were associated with elevated rates of all child disruptive behavior problems. Low levels of warm involvement were particularly characteristic of parents of children who showed elevated levels of oppositional behaviors. Physically aggressive parenting was linked more specifically with child aggression. In general, parenting practices contributed more to the prediction of oppositional and aggressive behavior problems than to hyperactive behavior problems, and parenting influences were fairly consistent across ethnic groups and sex. PMID- 10693031 TI - Fears, worries, and scary dreams in 4- to 12-year-old children: their content, developmental pattern, and origins. AB - Investigated anxiety symptoms in normal school children 4 to 12 years of age (N = 190). The percentages of children reporting fears, worries, and scary dreams were 75.8, 67.4, and 80.5%, respectively, indicating that these anxiety symptoms are quite common among children. Inspection of the developmental pattern of these phenomena revealed that fears and scary dreams were common among 4- to 6-year olds, became even more prominent in 7- to 9-year-olds, and then decreased in frequency in 10- to 12-year-olds. The developmental course of worry deviated from this pattern. This phenomenon was clearly more prevalent in older children (i.e., 7- to 12-year-olds) than in younger children. Furthermore, although the frequency of certain types of fears, worries, and dreams were found to change across age groups (e.g., the prevalence of fears and scary dreams pertaining to imaginary creatures decreased with age, whereas worry about test performance increased with age), the top intense fears, worries, and scary dreams remained relatively unchanged across age levels. An examination of the origins of these common anxiety phenomena showed that for fears and scary dreams, information was the most commonly reported pathway, whereas for worry, conditioning experiences were more prominent. PMID- 10693032 TI - Family processes as resources for African American children exposed to a constellation of sociodemographic risk factors. Family Health Project Group. PMID- 10693034 TI - Concurrent and longitudinal correlates of depressive symptoms among low-income, urban, African American children. Family Health Project Research Group. PMID- 10693033 TI - Disentangling the impact of low cognitive ability and inattention on social behavior and peer relationships. Conduct Problems Prevention Re search Group. AB - Examined the shared and unique contributions of low cognitive ability and inattention to the development of social behavior problems and peer relationships of children at the time of school entry. Kindergarten and first-grade assessments of cognitive ability, inattention and prosocial and aggressive behavior were collected for a multisite, normative sample. Sociometric assessments of peer relationships were collected at the end of first grade. Cognitive ability and inattention both contributed to the prediction of social behavior and peer relationships. Low cognitive ability was particularly predictive of prosocial skill deficits, and social behavior mediated the relation between cognitive ability and social preference. Inattention predicted both prosocial skill deficits and elevated aggressive-disruptive behavior problems. Behavior problems partially mediated the relation between inattention and social preference. Identified subgroups of children with elevated levels of inattention or low cognitive ability showed different patterns of peer problems, with low acceptance characteristic of the low cognitive ability (only) group and high dislike ratings characteristic of the inattentive and inattentive/low-ability group. Implications are discussed for the design of early intervention and prevention programs. PMID- 10693035 TI - Proactive and reactive aggression in delinquent adolescents: relations to aggression outcome expectancies. AB - Investigated whether the relation between aggression and the tendency to expect positive outcomes for aggressive behavior is specific to the proactive subtype of aggression (as opposed to the reactive subtype). In a sample of 86 incarcerated adolescent boys ages 13 to 18, we measured outcome expectancies for aggression using audiotaped hypothetical vignettes. For each participant, staff members completed proactive and reactive aggression rating scales. Regression analyses revealed that the relation between aggression and outcome expectancies was indeed specific to proactive aggression. Furthermore, this finding was supported regardless of whether outcome expectancies were assessed using vignettes describing proactive-aggressive behavior or those describing reactive-aggressive behavior. We discuss these findings and argue that interventions to reduce proactive or reactive aggression should differ from each other by addressing the specific social cognitive processes involved in each type of aggression. PMID- 10693036 TI - Role of the working alliance in the treatment of delinquent boys in community based programs. AB - Examined the role of the working alliance in the treatment of delinquent boys in community-based residential programs, clarifying the relation between therapeutic process and behavioral change. Horvath and Greenberg's (1989) Working Alliance Inventory was used to assess the therapeutic alliance between youth and staff after 3 weeks in treatment and again after 3 months. Achenbach's (1991) Child Behavior Checklist (CBCL; Youth Self-Report [YSR] and Teacher Report Form [TRF]) and recidivism scores were used to assess treatment progress and outcome. Results indicated that a positive working alliance assessed after 3 months in treatment related to positive psychological changes and predicted lower rates of recidivism. Unexpectedly, a positive working alliance assessed early in treatment was associated with negative outcomes (increased internalizing and externalizing symptoms and higher rates of recidivism). This finding suggests that for some delinquent youth initially optimistic assessments may be prognostic of slow progress or treatment failure. PMID- 10693037 TI - Personal adjustment and perceptions of grandchild behavior in custodial grandmothers. AB - Examined parenting stress, depression, parenting satisfaction, and perceptions of child behavior in 35 custodial grandmothers seeking outpatient psychological services for their grandchildren ages 3 to 12. These clinic grandmothers reported significantly greater stress and depression, less parenting satisfaction, and more negative perceptions of their grandchildren's behavior than did 35 nonclinic custodial grandmothers. Also, similar to a comparison group of 35 clinic mothers and to the empirical literature on clinic parents, the clinic grandmothers' personal adjustment (i.e., stress and depression) was related significantly to their perceptions of their grandchildren's behavior. Thus, assessment of custodial grandmothers' adjustment, as well as grandchild behavior, in clinical settings appears warranted. PMID- 10693038 TI - Assessing peer network and dyadic loneliness. AB - Describes the Peer Network and Dyadic Loneliness Scale (PNDLS), a new scale designed to assess simultaneously children's loneliness at multiple levels of peer relationships. Specifically, this scale measures loneliness associated with (a) lack of involvement in a social network and (b) the absence of a close dyadic friendship. Employing a sample of 209 5th-, 6th-, and 7th-grade boys and girls, the psychometric properties, interscale correlations, and preliminary validity data for the new scale are examined. Analyses revealed good internal consistency and a pattern of relationships with other loneliness, friendship quality, mutual best friendship, and sociometric social preference variables supporting the validity of the new scale. PMID- 10693039 TI - Resilient and stress-affected adolescents in an urban setting. AB - Studied 185 seventh- and eighth-grade inner-city adolescents. Participants were categorized as low and high in exposure to stressors (stressful events or neighborhood disadvantage) and externally exhibited competence (self-, teacher, and school reports). We predicted that resilient (high-stress/high-competence) and stress-affected (high-stress/low-competence) youth would differ across three domains of hypothesized protective resources: internal resources (i.e., coping skills, perceived competence), familial support, and extrafamilial support. We also predicted that there would be an emotional cost to resilient youth in terms of experiencing internalizing problems (depression, anxiety). There were direct effects for stressor level on several protective resources; however, the hypothesized protective resources did not discriminate resilient from stress affected youth. Both Resilient and stress-affected youth experienced equivalent levels of internalizing symptoms, and these groups' scores were higher than those of low-stress participants. These results are possibly reflective of the effects of chronic stressors. PMID- 10693040 TI - On our way to integrated bioethics: clinical/organizational/communal. PMID- 10693041 TI - The postmodern prescription: an antidote to hard boundaries and closed systems in healthcare organizations. PMID- 10693042 TI - Organizational ethics: creating structural and cultural change in healthcare organizations. PMID- 10693043 TI - Making the most of disequilibrium: bridging the gap between clinical and organizational ethics in a newly merged healthcare organization. PMID- 10693044 TI - Integrated ethics: synecdoche in healthcare. PMID- 10693045 TI - All in the family--siblings but not twins: the relationship of clinical and organizational ethics analysis. PMID- 10693046 TI - Linking professional and economic values in healthcare organizations. PMID- 10693047 TI - Actively engaging organizational ethics in healthcare: four essential elements. PMID- 10693048 TI - Confidentiality as an organizational ethics issue. PMID- 10693049 TI - Organizational ethics and sentinel events: doing the right thing when the worst thing happens. PMID- 10693050 TI - The "impossible patient": organizational response to a clinical problem. PMID- 10693051 TI - Determination of organochlorine and organophosphorus pesticide residues in low moisture, nonfatty products using a solid phase extraction cleanup and gas chromatography. AB - A multiresidue solid-phase extraction (SPE) method for the isolation and subsequent gas chromatographic determination of organochlorine and organophosphorus pesticide residues in low-moisture, nonfatty products is described. Residues are extracted from samples with an acetonitrile/water mixture. Cleanup of the extract is performed using graphitized carbon black and anion exchange SPE columns, and analysis is performed by gas chromatography with Hall electrolytic conductivity and flame photometric detection. Recovery data was obtained by fortifying corn, oats and wheat with pesticides. The average recoveries were 79-123% for eight organochlorine and 51-122% for 28 organophosphorus pesticide residues. The limit of quantitation for chlorpyriphos was 0.05 ppm using the Hall electrolytic conductivity detector and < 0.005 ppm using the flame photometric detector. PMID- 10693052 TI - Fate of metolachlor under subirrigation in a sandy soil: a lysimeter study. AB - A three-year field lysimeter study was conducted to investigate the role of subirrigation systems in reducing the risk of water pollution from metolachlor (2 chloro-N-(2-ethyl-6-methlphenyl)-N-(2-methoxy-1-methylethyl)ace tamide). Nine large PVC lysimeters, 1 m long x 0.45 m diameter, were packed with a sandy soil. Three water table management treatments, i.e. two subirrigation treatments with constant water table depths of 0.4 and 0.8 m, respectively, and a free drainage treatment in a completely randomized design with three replicates were used. Corn (Zea mays L.) was grown in each lysimeter, and at the beginning of summer of each year metolachlor was applied, at the locally recommended rate of 2.75 kg a.i./ha. Soil and water samples were collected at different time intervals after each natural or simulated rainfall event. Metolachlor was extracted from these samples and analyzed using Gas Chromatography. Results obtained in this three year study, (1993-1995), lead to the conclusion that metolachlor is quite mobile since it leached to a depth of 0.85 m below the soil surface quite early in the growing season. Metolachlor concentrations decreased with depth as well as with time. The shallower water table in the 0.4 m subirrigation treatment showed less residues in the soil solution than that of other treatments. However, a mass balance study, supported by an independent laboratory investigation, shows that water table management, statistically, has no significant effect on the reduction of metolachlor residues in sandy soils. PMID- 10693053 TI - Sorption of imidacloprid and its metabolites on tropical soils. AB - The sorption of imidacloprid (1-[(6-chloro-3-pyridinyl)-methyl]-N-nitro-2 imidazolid-inimine ) (IMI) and its metabolites imidacloprid-urea (1-[(6-chloro-3 pyridinyl)-methyl]-2-imidazol-idinone) (IU), imidacloprid-guanidine (1-[(6-chloro 3-pyridinyl)-methyl]-4,5-dihydro-1H-imidazol-2-amine) (IG), and imidacloprid guanidine-olefin (1-[(6-chloro-3-pyridinyl)methyl]-1H-imidazol-2-amine) (IGO) was determined on six typical Brazilian soils. Sorption of the chemicals on the soil was characterized using the batch equilibration method. The range and order of sorption (Kd) on the six soils was IG (4.75-134) > or = IGO (2.87-72.3) > IMI (0.55-16.9) > IU (0.31-9.50). For IMI and IU, Kd was correlated with soil organic carbon (OC) content and CEC, the latter due to the high correlation between OC and cation exchange capacity (CEC) (R2 = 0.98). For IG and IGO, there was no correlation of sorption to clay, pH, OC or CEC due to the high sorption on all soils. Average Koc values were IU = 170, IMI = 362, IGO = 2433, and IG = 3500. Although Kd and Koc values found were consistently lower than those found in soils developed in non-tropical climates, imidacloprid and its metabolites were still considered to be slightly mobile to immobile in Brazilian soils. PMID- 10693054 TI - Monitoring of pesticide residues in a cotton crop soil. AB - This paper reports on the residues of methyl parathion (O,O-dimethyl O-4 nitrophenyl phosphorothioate), trifluralin (alpha, alpha, alpha-trifluoro-2,6 dinitro-N,N-dipropyl-p-toluidine), endosulfan [(1, 4, 5, 6, 7, 7-hexachloro-8, 9, 10-trinorborn-5-en-2, 3-ylenebismethylene) sulfite] and dimethoate (O, O-dimethyl S-methylcarbamoylmethyl phosphorodithioate) in a cotton crop soil. Soil samples (0-15 cm) were collected at different periods from the cotton crop farm and subjected to Soxhlet extraction. The extracted material was analysed after clean up by a HP5890 II gas chromatograph equipped with a 63Ni electron-capture detector (ECD-63Ni) and fitted with a 25 m x 0.2 mm i.d. fused silica capillary column [Ultra-2 (5% phenylmethyl polysiloxane)]. The recoveries of the pesticide residues from the spiked control soil were determined after Soxhlet extraction and C18 cartridges clean-up by using radiotracer techniques with the corresponding 14C-pesticides. The results show that in the cotton crop soil the pesticide residues under study were present in the range of 0.1 to 0.4 mg.kg-1. Endosulfan was found to be rapidly degraded in the soil and formed a sulfate metabolite. PMID- 10693055 TI - Photocatalytic oxidation of xenobiotics in water with immobilized TiO2 on agitator. AB - A novel photocatalytic oxidation reactor, using Degussa P-25 TiO2 as a stationary phase with a thickness of 1.5-2.0 um on the blades of agitator, was developed to study the photocatalytic oxidation of xenobiotics. Particularly in this device, separation of photocatalyst from the purified water after oxidation reaction was not necessary, and no other aeration equipment was required to supply oxygen. To examine the efficiency of this device, photocatalytic degradation of xenobiotic organics such as carbofuran was studied as an example. Results indicated that carbofuran could be degraded completely with mineralization efficiency of 20% after 6 hours of oxidation under the imposed conditions. The mineralization rate of carbofuran was found to follow the pseudo-first order reaction kinetics. Moreover, the rate constant of mineralization was found to be proportional to TiO2 film area and the square root of UV light intensity. These results implied the mineralization efficiency of carbofuran could be improved through increasing TiO2 film area and UV light intensity. Accordingly, this novel device showed potential application for degrading xenobiotics in water. PMID- 10693056 TI - Effect of dimethoate on the immune system of female mice. AB - The functional status of the immune system of female mice exposed to a single oral dose of dimethoate (16 mg/kg) was evaluated by assessing cell mediated and humoral immune responses, in addition to the effect of dimethoate on spleen and body weights after different time intervals. The data showed that dimethoate caused a time-depended decrease in spleen weights in the absence of a change in body weights. The immunologic effect of dimethoate to female mice produced a dose dependent decrease in the number of the rosette forming cells (total and active erythrocyte rosette). The ability of splenocytes to proliferation in response to mitogens; phytohemagglutinin (PHA) for T cell and lipopolysaccharide (LPS) for B cell were significantly decreased at the different times. As compared to control, a significant decrease in serum total immunoglobulins (Ig) and IgM was found, while IgG was non-significant deceased. Results of this study also revealed that dimethoate caused a significant decrease in the number of plaque forming cell (PFC/10(6) splenocytes) in a time dependent manner. PMID- 10693057 TI - Investigation of metal ions binding of humic substances using fluorescence emission and synchronous-scan spectroscopy. AB - The binding site interactions of IHSS humic substances, Suwannee River Humic Acid, Suwannee River Fulvic Acid, Nordic Fulvic Acid, and Aldrich Humic Acid with various metals ions and a herbicide, methyl viologen were investigated using fluorescence emission and synchronous-scan spectroscopy. The metal ions used were, Fe(III), Cr(III), Cr(VI), Pb(II), Cu(II) and Ni(II). Stern-Volmer constants, Ksv for these quenchers were determined at pH 4 and 8 using an ionic strength of 0.1 M. For all four humic substances, and at both pH studied, Fe(III) was found to be the most efficient quencher. Quenching efficiency was found to be 3-10 times higher at pH 8. The bimolecular quenching rate constants were found to exceed the maximum considered for diffusion controlled interactions, and indicate that the fluorophore and quencher are in close physical association. Synchronous scan spectra were found to change with pH and provided useful information on binding site interactions between humic substances and these quenchers. PMID- 10693058 TI - Glutamine synthetase and protease enzyme activities and growth response of ruminal bacterium Prevotella ruminicola strain B(1)4 to nitrogen source and concentration. AB - The objective of this study was to determine the effects of varying nitrogen sources and concentrations upon glutamine synthetase and protease activities in Prevotella ruminicola strain B(1)4. Based on growth response it appears that ammonium chloride or pepticase limited P. ruminicola becomes nitrogen limited when nitrogen concentration is at 0.5 mM. However, when casein was provided as the sole source of nitrogen P. ruminicola becomes nitrogen limited at 2.5 mM. Glutamine synthetase activity was measured from mid-log phase cells grown in either nitrogen-limited or non-limited conditions. No activity was detectable in the non-limited treatments. However, in the N-limited treatments, pepticase had the highest activity (20.76 units), followed by ammonium chloride (18.72 units) and casein (14.42 units). Protease activity assays indicated that nitrogen limited cultures had higher proteolytic activity than non-limited cultures. Moreover, these activities appeared to follow the same response pattern as the previously observed glutamine synthetase activities. The results of this study indicate that P. ruminicola strain B(1)4 protease activity may be influenced by nitrogen concentration such that activity increases when nitrogen availability decreases. PMID- 10693059 TI - Preventing neural tube defects: a major success story, with a chapter yet to be written. PMID- 10693060 TI - Interventions to increase breast screening uptake: do they make any difference? AB - BACKGROUND: Breast screening has an important role in improving survival from breast cancer through early detection and treatment. Increasing uptake of screening in areas of low uptake is important in improving the effectiveness of the national screening programme. This review looks at which initiatives to boost uptake have been successful. OBJECTIVE: To evaluate the effectiveness of the different interventions to increase breast screening uptake. METHOD: A systematic review of interventions to promote breast screening uptake was undertaken. Studies were included if uptake was used as an outcome measure of the intervention and if relevant to the UK screening programme. RESULTS: Twenty eight studies were found among 25 citations. Interventions were grouped into "person directed", "system directed", "social network directed", and "multistrategy" categories. Most were person directed. These interventions were more likely to be effective in boosting uptake, be simple in design, and to have been evaluated by a randomized trial design. Evidence of effectiveness in the other groups is limited both by the number of studies and the study designs. A summary of the interventions reviewed is presented. CONCLUSIONS: Simple, brief, and effective interventions exist to boost breast screening uptake. More complicated approaches are not necessarily any more effective. These findings also have implications for other population based screening programmes of the future. In inner city areas the best approach to raising uptake rates is likely to be multistrategy. PMID- 10693061 TI - Quantifying the decline in the birth prevalence of neural tube defects in England and Wales. AB - In England and Wales there has been a large decline in the birth prevalence of neural tube defects (NTDs) from the early 1970s (reported rates of about 3.2 per 1000 births) to the present (0.1 per 1000 births in 1997). The reported number of terminations of NTD pregnancies increased from 0.02 per 1000 in 1970 to 0.66 per 1000 in 1997, much too small an increase to explain the decline in NTD births. Some underreporting of NTD terminations is recognised. We estimated its extent using 1976-80 data on reported central nervous system (CNS) defect terminations and NTD births in an analysis in which the true total number of NTD pregnancies during this short period was assumed to have either remained constant or else to have been changing by a constant amount per year. The estimate was that 56% of NTD terminations were not reported as such and this fitted the data well. In 1997 the estimated birth prevalence of NTDs was 0.14 per 1000 births, a fall of 96% since 1970. This 96% was apportioned as 40% due to antenatal screening and termination of pregnancy and 56% due to a decline in incidence. Over the period 1970-97 there was an increase in dietary folate, and this will have at least in part caused the decline in incidence of NTDs. PMID- 10693062 TI - Current status of neonatal screening in China. PMID- 10693063 TI - Comparison of the CAPAS and Ewing tests for screening of hearing in infants. AB - OBJECTIVE: To study the similarities and differences between the non-automated labour intensive Ewing hearing test and the less labour intensive automated CAPAS (Compact Amsterdam Paedo-Audiometrical Screening) hearing test. SETTING: A multicentre study in which all the children born in the eastern part of the Netherlands between 1 January 1996 and 1 April 1997 were routinely screened for hearing impairment at 9 months of age. METHODS: Differences and similarities between the two methods were described for the proportion of children who failed every test, the percentage of referred children, the yield of bilateral and unilateral otitis media with effusion (OME), the positive predictive value of the third test result, and the yield of persistent OME after 4-6 months' follow up at an ENT department. RESULTS: 12,603 infants were screened with the CAPAS test and 17,496 with the Ewing test. There were differences between the CAPAS and Ewing tests respectively in the proportions of children lost to follow up (10.1% v 15.2%), the proportions of children referred diagnosed with OME (59% v 81%), the yield of bilateral otitis media with effusion (2.4% v 3.0%), and the yield of persistent OME after 4-6 months' follow up (1.1% v 1.6%). CONCLUSIONS: The CAPAS test is more practical than the Ewing test, but the non-automated Ewing test seems to be more reliable and valid for detecting conductive hearing loss. PMID- 10693064 TI - Should genetic analysis in newborn screening and a heterozygote test for hyperphenylalaninaemia be recommended? An Italian study. AB - OBJECTIVE: To determine whether the introduction of genetic analysis for phenylalanine hydroxylase (PAH) deficiency into regional screening programmes can be supported by the benefit-cost ratio. METHOD: Tests for the genetic PAH locus were carried out in 151 patients with hyperphenylalaninaemia originally from all of the Italian regions. PAH mutations were identified by extraction of genomic DNA from leucocytes (whole blood in EDTA), PAH exon amplification was determined by polymerase chain reaction, restriction enzyme analysis was carried out for some recognised mutations, and DNA sequence analysis for the other mutations. RESULTS: It was found that the eight most common mutations in the population accounted for 49% of the mutant alleles, which is well below the required standard for effective population screening (90%). CONCLUSIONS: Genetic screening for PAH deficiency in Italy does not increase the sensitivity of the methodology and the benefit-cost ratio, and thus provides no advantage, particularly as the correlation between genotype and the metabolic phenotype needed to optimise dietary intervention is still being studied. PMID- 10693065 TI - The target plot: a new way of displaying the performance of a screening test. AB - OBJECTIVES: To produce a graphical method to represent the performance of a screening test that illustrates the prevalence of the disorder being screened for, as well as its discriminatory potential. CONCEPT: A target plot was constructed in which the risk of the disorder is represented by a series of concentric circles of constant risk (isorisks) equivalent to specified false positive rates, with the highest risk in the centre and lower risks spreading outwards towards the circumference. Dots were drawn to represent cases of the disorder; these were of a size such that their total area as a proportion of the area of the whole target plot equalled the prevalence of the disorder in the screened population. The discriminatory power of the test was seen as the clustering of dots around the centre or bull's eye of the target. The detection rate could be estimated as the proportion of dots which fell within the isorisk corresponding to a specified false positive rate. APPLICATION: The target plot was applied to second trimester antenatal screening for Down's syndrome using different combinations of screening markers, and also to screening for ischaemic heart disease using protein components of cholesterol (apolipoproteins A I and B and Lp(a) lipoprotein), systolic blood pressure, and smoking status. DISCUSSION: The efficacy of the different methods of screening for Down's syndrome is readily apparent using the target plot, as is the poorer performance of screening for ischaemic heart disease. CONCLUSIONS: The target plot is a simple and quantitative way of displaying the performance of a screening test that may be useful in teaching and educational material. PMID- 10693066 TI - High mammographic breast density and its implications for the early detection of breast cancer. AB - OBJECTIVES: Women with high mammographic breast density are at increased risk of breast cancer. This study explores whether these women should receive intensified screening (more frequent screening or screening with alternative techniques that increase the length of the preclinical detectable phase) to reduce further breast cancer mortality. METHODS: Mathematical models were used to estimate the effects of intensified screening in women with high breast density. The effects were expressed as a reduction in the number of interval cancers. RESULTS: If women with > 25% breast density (comprising about one fifth of all women) are screened annually instead of biennially, an 18% reduction in the total number of interval cancers can be expected. Screening these women with alternative screening techniques biennially may produce the same reduction, provided that these techniques double the mean lead time. CONCLUSIONS: By screening women with dense breasts more intensively, many more breast cancers can theoretically be detected at an early stage. The results provide an early indication of what may be expected from screening strategies. Next, cost-benefit analyses are needed. PMID- 10693067 TI - Identifying and screening patients at high risk of colorectal cancer in general practice. AB - OBJECTIVES: To determine the feasibility and acceptability of selecting patients at risk of colorectal cancer by taking family histories by means of a postal questionnaire. To determine if this information could be translated into simple risk categories to guide subsequent management. SETTING: Patients aged between 30 and 69 years inclusive, registered with a mixed suburban and rural training general practice in south west England. METHOD: A postal questionnaire survey seeking demographic information and family history of colorectal cancer was sent to all eligible patients. Personal risk of colorectal cancer was stratified according to predetermined criteria. Risk assessment was modified if necessary after the general practitioner conferred with a geneticist. Patients were subsequently offered colonoscopy (high risk) or faecal occult blood testing (intermediate risk). RESULTS: Response to the questionnaire was 84.7%. 250 patients had a family history of colorectal cancer, of whom 52 were assigned to the high risk group, 104 to the intermediate group, and 94 to the low risk group. The geneticist reassigned five intermediate risk patients to the high risk group. Of 27 patients who had a colonoscopy, two were found to have an adenocarcinoma and a further two adenomatous polyps. In the group given faecal occult blood testing, two patients presented with colorectal cancer before being screened. CONCLUSIONS: A postal questionnaire is feasible and acceptable for the collection of information about a family history of colorectal cancer from patients in general practice. The personal risk of developing the disease according to standard criteria can be estimated and then managed by a simple protocol. PMID- 10693068 TI - Implementation of screening as a public health policy: issues in design and evaluation. AB - OBJECTIVE: To propose principles of design and measures of effect for cancer screening as a public health policy. MATERIAL: Finnish routine screening programme with mammography. DESIGN: Evaluation of mortality from breast cancer by before-after time trends among Finnish women, by geographical (by municipalities) comparisons, and by intention to screen at an individual level with individual controls who were cluster randomised and matched for age. OUTCOME MEASURES: Standardised mortality ratios (SMRs) from breast cancer (total), SMRs from breast cancers diagnosed during the screening programme (refined), deaths prevented, prolongation of life for each breast cancer detected, for each death prevented, for each compliant woman, for each screen, and for each invitation. Relative prolongation of life--that is, time gained versus time spent. RESULTS: SMRs only at an individual level with deaths from cancers diagnosed during the screening programme and individually selected controls showed 24% protection, whereas the other SMR measures were too crude or biased because of dilution and selection. Prolongation of life varied from 15 years for each death prevented to two days for each woman screened, with approximation for the prolongation relative to time spent of 3 to 1. CONCLUSIONS: A public health policy should be introduced gradually. Those not covered immediately by the policy serve as controls and they should be randomly allocated. The most relevant outcome measure is prolongation of life, and for public health purposes it should be given per unit of intervention, such as screen or invitation, and also related to the time spent for the intervention. Such gains are often small in a Western society, which implies that medicine, including research, should focus more on other aspects of life than length. PMID- 10693069 TI - Assessment of complete blood count variations among workers exposed to low levels of benzene. PMID- 10693070 TI - Education for the practice of occupational medicine: knowledge, competence, and professionalism. AB - The multistep process of education is delineated by the sequential phases: (1) Knowledge Transfer, (2) Competence Development, and (3) Professional Inculcation. The realities of practice modes and curricular time constraints are important determinants of the breadth and depth of the information provided in the Knowledge Transfer process. Accordingly, it is proposed that Phase 1, the Knowledge Acquisition Process, be organized into two components: (1) Core Knowledge, requiring both significant breadth and depth; and (2) Augmentive Knowledge, providing wide breadth and appropriate but variable depth. This curricular organizing proposition recognizes that: (1) the wide breadth of multiple stores of knowledge inherent in the practice of PM and EOM considerably exceeds many other medical specialities; (2) the duration of training is inherently shorter; and (3) its practitioners generally operate as members of teams consisting of other professionals (e.g., attorneys, engineers, business administrators, industrial hygienists, sociologists, psychologists). Obviously, it is unreasonable to expect the members of such teams to each have comparable depth and breadth of knowledge. A broad knowledge base, implicit in Augmentive Knowledge, provides the capacity for recognition, understanding, and application of capabilities brought by other professionals. Facilitating communications between team members, each possessing a broad knowledge base, enhances the effectiveness of the knowledge, competence, and professionalism of collaborative efforts. Phase 2, Competency, consists of the coherent integration of multiple stores of information applicable to the management of a clearly defined task with a clearly measurable outcome. The accomplishment of true competency is not based on the simple possession of multiple stores of knowledge; rather it depends on the facility and effectiveness with which information bases are marshaled, integrated, and communicated. Clearly, the effectiveness of this process increases with its interaction; it is unreasonable to expect a significant degree of competence immediately upon graduation from a training program. Phase 3, Professionalism, and its basic ethos provides the governing context for the sound application of competencies. Although it is difficult to teach, only with its accomplishment can the educational process be considered whole, albeit never complete. PMID- 10693071 TI - Personal exposure to atmospheric polycyclic aromatic hydrocarbons in a general adult population and lung cancer risk assessment. AB - Personal exposure to nine particulate-phase atmospheric polycyclic aromatic hydrocarbons (PAHs) was assessed among adult non-smoking volunteers in the Grenoble, France, metropolitan area. Using Toxic Equivalency Factors, the associated total atmospheric PAHs lifelong cancer risk was estimated. For 48 hours continuously, 38 subjects without specific occupational exposure to combustion sources carried a PM2.5 particles personal exposure monitor while at home, at work, commuting, or involved in other activities. One phase of the study took place in summer; a second in winter. The monitor set was composed of a pump with an airflow of 4 L.mn-1, a 2.5-micron cyclone, and Teflon filters. The PAH concentrations were determined on seven PM2.5 filters by using high performance liquid chromatography with fluorimetric detection. The predominant PAHs are fluoranthene and indeno pyrene. According to the compound, the personal exposure estimates ranged from 0.13 to 1.67 ng/m3 (yearly means). The average benzo(a) pyrene value is 0.67 ng/m3 (95% confidence interval = 0 to 2.1 ng/m3). Winter exposures were 3 to 25 times greater than summer exposures. The total PAHs lung cancer lifelong risk is 7.8 10(-5) and is driven by exposure to benzo(a) pyrene. Although these risk estimates are 2 to 3 orders of magnitude lower than those associated with specific occupational exposures in the coal or smelter industries, they are of public health concern because they are spread over large urban populations. Further personal exposure studies in adult or children populations are needed. PMID- 10693072 TI - Association of physical activity at work with mortality in Israeli industrial employees: the CORDIS study. AB - The objective of this study was to evaluate the association of physical activity at work with the risk of all-cause cardiovascular disease and cancer mortality. The cohort consisted of 3488 male, Israeli, industrial employees who participated in an 8-year follow-up study. During this period 129 deaths were recorded: 54 from cardiovascular disease, 47 from cancer, and 28 from other causes. Physical activity at work was assessed at entry on a 4-point scale (none, light, medium, and high). Potential confounding demographic, anthropometric, and socioeconomic variables, and health habits including leisure time physical activity were accounted for. We found that the hazard ratio of all-cause mortality in workers with a high physical workload was 1.82 (95% confidence interval, 1.18 to 2.81) compared with workers having a low workload. A similar trend was noted for cardiovascular disease and cancer mortality. We concluded that a high physical workload is associated with increased mortality rates. Future studies should differentiate between leisure time and work time physical activity. PMID- 10693073 TI - Melatonin metabolite levels in workers exposed to 60-Hz magnetic fields: work in substations and with 3-phase conductors. AB - Melatonin suppression by 50/60-Hz magnetic fields represents a plausible biological mechanism for explaining increased health risks in workers. Personal exposure to magnetic fields and ambient light, and excretion of the melatonin metabolite 6-hydroxymelatonin sulfate (6-OHMS), were measured over 3 consecutive workdays in electric utility workers. There was a magnetic field-dependent reduction in adjusted mean nocturnal and post-work 6-OHMS levels among men working more than 2 hours per day in substation and 3-phase environments and no effect among those working 2 hours or less. No changes were observed among men working in 1-phase environments. The results suggest that circular or elliptical magnetic field polarization, or another factor linked to substations and 3-phase electricity, is associated with magnetic field induced melatonin suppression in humans. PMID- 10693074 TI - Decrease of suppressor-inducer (CD4+ CD45RA) T lymphocytes and increase of serum immunoglobulin G due to perceived job stress in Japanese nuclear electric power plant workers. AB - To clarify the effects of perceived job stress on the immune system, a cross sectional study was conducted in 116 male Japanese workers of a nuclear electric power plant (age, 20 to 39; mean, 31 years). Perceived job stress, i.e., psychological job demand, job control, worksite social support, and job strain, was assessed by means of the Japanese version of the Job Content Questionnaire. The job strain score was calculated as the ratio of the job demand score to the job control score. Blood samples were taken from all workers, and numbers of T and natural killer cell subpopulations, B lymphocytes, total lymphocytes and white blood cells, and serum concentrations of immunoglobulins (IgG, IgM, IgA, IgE and IgD) in their blood were measured. The workers were divided into higher and lower strain groups according to their job strain scores. The number of CD4+ CD45RA+ T lymphocytes in the higher strain group having the job strain score of 0.5 or more (41 workers) was significantly smaller than that in the lower strain group having the score of less than 0.5 (75 workers). In contrast, the serum IgG concentration in the former group was significantly higher than that in the latter group (analysis of covariance with age and smoking as covariates). Also, the numbers of total CD4+ T and total T (CD3+) lymphocytes and of white blood cells in the former group were significantly smaller than those in the latter group. After controlling for age and smoking by the partial correlation coefficient in all 116 workers, the number of CD57+ CD16+ natural killer cells was inversely correlated with job demand and with job strain; the number of CD8+ T lymphocytes was positively correlated with worksite social support; and serum IgG and IgM concentrations were positively correlated with job strain. It is suggested that higher job strain decreases the number of CD4+ CD45RA+ T lymphocytes in male Japanese workers but increases serum IgG concentrations. PMID- 10693075 TI - Association between aminolevulinate dehydrogenase genotype and blood lead levels in Taiwan. AB - This study was designed to evaluate the association between the aminolevulinate dehydrogenase (ALAD) genotype and blood lead levels in a general population environmentally exposed to lead. This study population of 660 subjects was secondarily sampled from the 3000 random samples of Taiwanese general population to study the distribution of blood lead levels in the Taiwanese population. A simple assay based on the polymerase chain reaction-restriction fragment length polymorphism technique was used to determine the genotype of the ALAD gene. This study found that most of the Taiwanese population was ALAD 1-1 (95.4%). Only 4.6% (30 subjects) of population were found to be 1-2 or 2-2. It has been hypothesized that the ALAD2 allele is associated with increased absorption of lead. This study found that individuals with ALAD2 alleles had 20% higher blood lead levels than persons with ALAD1 alleles (7.83 +/- 5.95 vs 6.51 +/- 5.03 micrograms/dL). However, the difference was not statistically significant, even after adjustment for other risk factors of environmental exposure. The result supports the previous finding that individuals with ALAD2 allele had higher blood lead levels. The small sample size and large amount of variation in our study may account for the insignificant association. PMID- 10693076 TI - Work-related deaths in West Virginia from July 1996 through June 1999: surveillance, investigation, and prevention. AB - The National Institute for Occupational Safety and Health's Fatality Assessment and Control Evaluation model is used to identify and describe work-related deaths in West Virginia. Through a statewide surveillance network, this model identifies work situations at high risk for fatal injury, investigates selected causes (falls, machinery-related, and logging), and formulates and disseminates prevention strategies to reduce the frequency and impact of those injuries. A total of 163 persons died from work-related injuries from July 1996 through June 1999. Ninety-three percent were male, the mean age was 42, and 80% were West Virginia residents. Fatalities occurred most frequently in the transportation/public utilities (32), manufacturing (24), construction (23), and mining (23) industries. Extension of Fatality Assessment and Control Evaluation methodology to nonfatal injuries may contribute to a clearer understanding of the causes of these traumatic incidents and help to develop better prevention measures. PMID- 10693077 TI - A framework for addressing health issues in or near a manufacturing facility. AB - Clustering of health events in or around industrial facilities sometimes leads to worker and community concerns that plant management or local health professionals must address. We provide an eight-step process to deal with these concerns systematically. We emphasize the use of good scientific practices with managerial oversight for effective worker and community communication. This process is directed to plant management and the local health professional and emphasizes the practical aspects of the investigation. PMID- 10693078 TI - The pervasiveness of the illness suffered by workers seeking compensation for disabling arm pain. AB - Disability from work-related arm pain has become prevalent in several countries in recent years. Many of these individuals present with chronic musculoskeletal symptoms that, for lack of a more specific diagnosis, are often labeled as a repetitive strain injury or cumulative trauma disorder. Indemnity for such conditions can be contentious; many of these sufferers are involved in litigation in their quest for financial compensation for temporary or permanent disability. This article describes our experience with 103 patients referred to a Health Reference Center for Workers for the management of repetitive strain injury. Their illness is far more global than the work-related arm pain that such labeling implies. From the total group, 73 fulfilled the American College of Rheumatology Criteria for the Classification of Fibromyalgia Syndrome. This means that they were suffering pain above and below the diaphragm, far from the arm pain for which they were referred. These 73 patients were clinically and psychologically indistinguishable from 165 patients followed in our clinic at the Federal University of Sao Paulo, Rheumatology Division, who also fulfilled these criteria but did not consider their illness work-related. This observation calls for longitudinal investigations that might offer insights as to whether the more global aspects of the illness are antecedent, coincident, or confounding aspects of the illness experience labeled repetitive strain injury or cumulative trauma disorder. PMID- 10693079 TI - Injury and employment patterns among Hispanic construction workers. AB - This article describes non-fatal injuries among Hispanic construction workers treated at an emergency department from 1990 to 1998. Medical and interview data were analyzed to evaluate and explain the workers' apparently inflated risk of injury. The majority of the injured Hispanic workers were employed in the less skilled trades. Compared with other injured workers, Hispanics had a higher proportion of serious injuries and were disadvantaged in terms of training and union status. With the exception of union status, these differences largely disappeared after controlling for trade. The physical, financial, and emotional consequences were more apparent 1 year later for injured Hispanics, even after controlling for trade. These observations suggest that minority status is a predictor of trade and that trade is a predictor of injury risk. In addition to reducing injury hazards, interventions should address the limited employment and union membership options that are available to minority workers in the construction industry. PMID- 10693080 TI - Association of pesticide safety knowledge with beliefs and intentions among farm pesticide applicators. AB - Although a number of health hazards associated with pesticide exposure have been well documented, relatively little is known about the knowledge and health beliefs that may influence pesticide handling. This study measured knowledge levels concerning pesticide safety and precautionary handling among applicators and examined relationships between knowledge scores and intentions to use handling precautions, perceptions of pesticide safety peer norms, and perceived self-efficacy to prevent personal exposure. Telephone interviews were conducted with a randomly selected sample of 164 dairy farmers who were pesticide applicators residing in Wisconsin (response rate = 77.4%). The percentage of correct responses to 18 knowledge items ranged from 100% to 45.7%. Knowledge levels were positively related to intentions, beliefs, and self-efficacy regarding use of personal protective gear but were not significantly related to risk perceptions and peer norms concerning pesticide safety. PMID- 10693081 TI - A cohort mortality study among gas generator utility workers. AB - An earlier cohort study tracked the mortality experience through 1988 of male employees at five utility companies in the United States. Workers employed by the Pacific Gas and Electric Company (PG&E) were part of that study, but results for PG&E employees overall or for those involved in gas generator plant operations where hexavalent chromium compounds were used in open and closed systems from the 1950s to early 1980s were not reported. To evaluate risk of lung cancer and other diseases, a cohort of 51,899 PG&E male workers was followed for mortality from 1971 through 1997. Observed numbers of deaths were compared with those expected based on rates in the general California population, with standardized mortality ratios (SMR) and corresponding 95% confidence intervals (CI) calculated for the total cohort and for subsets defined by potential for gas generator plant exposure. A total of 10,591 deaths were observed, a number significantly less than expected (SMR, 0.89; 95% CI, 0.87 to 0.91). No significant excesses of total or specific cancers were observed, with SMR typically near or below 1.0. Lung cancer mortality in the entire cohort was close to expected (SMR, 0.98; 95% CI, 0.92 to 1.05), with no excess detected among persons who worked (SMR, 0.81; 95% CI, 0.35 to 1.60) or trained (SMR, 0.57; 95% CI, 0.12 to 1.67) at gas generator facilities. Furthermore, risk of lung cancer did not increase with increasing duration of employment or time since hire. The study thus provides no evidence that occupational exposures at PG&E facilities resulted in increased risk of lung cancer or any other cause of death. The results indicate that any chromium exposures were of insufficient magnitude to result in increased risk of lung cancer. PMID- 10693083 TI - Total ankle replacement revisited. AB - The surgical treatment of painful, end-stage ankle arthritis includes ankle arthrodesis and total ankle replacement. In the past decade, total ankle replacement has become a viable alternative to ankle arthrodesis. Modern implant designs either involve a syndesmosis fusion and resurfacing of the medial and lateral recesses of the ankle joint or the use of a 3-component, mobile bearing implant. In limited clinical series, the early results of both these prosthetic design approaches are encouraging. In selected patients, ankle arthroplasty is an effective approach to relieving pain and improving function. The purposes of this paper are to review the clinical results from total ankle replacement and ankle arthrodesis; discuss indications, contraindications, design features, postoperative rehabilitation, and initial results for the major current total ankle designs; and present concepts for future total ankle development. In particular, this article explores the advantages and concerns with 2 prevalent but different design approaches. It also discusses future directions for total ankle replacement. PMID- 10693082 TI - Neonatal thyroxine level and perchlorate in drinking water. AB - Environmental contamination of drinking water has been observed for perchlorate, a chemical able to affect thyroid function. This study examines whether that exposure affected the thyroid function of newborns. Neonatal blood thyroxine (T4) levels for days 1 to 4 of life were compared for newborns from the city of Las Vegas, Nevada, which has perchlorate in its drinking water, and those from the city of Reno, Nevada, which does not (detection limit, 4 micrograms/L [ppb]). This study is based on blood T4 analyses from more than 23,000 newborns in these two cities during the period April 1998 through June 1999. No difference was found in the mean blood T4 levels of the newborns from these two cities. Drinking water perchlorate levels measured monthly for Las Vegas ranged during this study period from non-detectable for 8 months to levels of 9 to 15 ppb for 7 months. Temporal differences in mean T4 level were noted in both cities but were unrelated to the perchlorate exposure. This study was sufficiently sensitive to detect the effects of gender, birth weight, and the day of life on which the blood sample was taken on the neonatal T4 level, but it detected no effect from environmental exposures to perchlorate that ranged up to 15 micrograms/L (ppb). PMID- 10693084 TI - Posterior tibial tendon dysfunction. AB - Posterior tibial tendon dysfunction is the most common cause of acquired flatfoot deformity in adults. Although this term suggests pathology involving only the posterior tibial tendon, the disorder includes a spectrum of pathologic changes involving associated tendon, ligament, and joint structures of the ankle, hindfoot, and midfoot. Early recognition and treatment is the key to prevention of the debilitating, long-term consequences of this disorder. Conservative care is possible in the earliest stages, whereas surgical reconstruction and eventually arthrodeses become necessary in the latter stages. The purpose of this article is to review the symptoms, physical examination, radiological examination, classification, and treatment of posterior tibial tendon dysfunction. PMID- 10693085 TI - Differential diagnosis of calf pain and weakness: flexor hallucis longus strain. PMID- 10693086 TI - Effects of a tendo-Achilles lengthening procedure on muscle function and gait characteristics in a patient with diabetes mellitus. AB - STUDY DESIGN: Case report with repeated measures. OBJECTIVES: To describe the effects of a tendo-Achilles lengthening (TAL) and total contact casting (TCC) on wound healing, motion, plantar pressure, and function in a patient with diabetes mellitus, peripheral neuropathy, neuropathic ulcer, and limited dorsiflexion range of motion (DFROM). BACKGROUND: Limited DFROM has been associated with increased forefoot pressures and skin breakdown. A TAL was expected to increase DFROM and reduce forefoot pressures during walking, but the influence on muscle performance and function was unknown. METHODS AND MEASURES: The patient was a 42 year-old man with a 20-year history of type 1 diabetes (NIDDM) and a recurrent neuropathic plantar ulcer. Outcome measures were DFROM, isokinetic plantar flexor muscle peak torque, in-shoe and barefoot peak plantar pressure, physical performance test (PPT) score, and peak ankle and hip moments during walking obtained from an automated gait analysis. All tests were completed pre-TAL, 8 weeks post-TAL (after immobilization in a TCC), and 7 months post-TAL. RESULTS: The wound healed in 40 days. The TAL resulted in a sustained increase in DFROM (0 to 18 degrees). Plantar flexor peak torque was reduced by 21% 8 weeks after the TAL compared with the torque before surgery but recovered fully at 7 months. Seven months following TAL, in-shoe forefoot peak plantar pressure was reduced by 55%, barefoot pressure decreased by 14%, PPT score increased by 24%, peak ankle plantar flexor moment remained decreased by 30%, and the peak hip flexor moment increased by 41% during walking. CONCLUSION: For this patient, a TAL resulted in short-term deficits in peak plantar flexor torque, but a 7-month follow-up showed improvements in ankle DFROM, walking ability, and a decrease in forefoot in-shoe peak plantar pressure. PMID- 10693087 TI - Factors associated with ulceration and amputation in the neuropathic foot. AB - The purpose of this paper is to review selected literature on the relationship of neuropathy and other related factors in foot ulceration and lower extremity amputation. There is strong evidence that sensory loss and mechanical stress are the primary cause of foot ulceration and common factors in the pathway to lower extremity amputation. Foot stress results from extrinsic factors such as footwear and intrinsic factors such as deformity and limited joint mobility. Understanding the interplay of these factors is valuable in identifying persons whose feet are at risk, effectively preventing and treating foot ulcerations and ultimately preventing lower extremity amputation. PMID- 10693088 TI - Against the myth of evidence-based practice. PMID- 10693089 TI - Measurement of posterior shoulder tightness. PMID- 10693090 TI - The perinatal history of the Section on Perinatal Pediatrics. PMID- 10693091 TI - Section on Perinatal Pediatrics: the 1980s to the 1990s. PMID- 10693092 TI - On the 25th anniversary of the Section on Perinatal Pediatrics: historical foresight. PMID- 10693093 TI - Moral and ethical dilemmas in critically ill newborns: a 20-year follow-up survey of Massachusetts pediatricians. AB - OBJECTIVE: To replicate the 1987 survey, referring to the original 1977 study, regarding opinions about treatment for critically ill neonates. STUDY DESIGN: A long-term follow-up survey of American Academy of Pediatrics Massachusetts membership, maintaining the 1987 instrument, was initiated. RESULTS: A notable demographic shift in respondents from a majority of male practitioners in 1977 (89.6%), to 73% in 1987, to more equal numbers of men and women in 1997 (55% and 45%, respectively; p < 0.001; 1987 vs 1997) was apparent. Pediatricians' attitude changes over the 20-year period were relatively modest and were statistically associated with active medical intervention. In 1997, 75% of respondents rejected review committees as mediators, a marked change from 1987. Regardless of healthcare maintenance organization affiliations, 95% indicated that restrictive fiscal policies would not affect decision-making. CONCLUSION: This study indicates stability and consensus in pediatricians' attitudes toward active intervention for critically ill neonates compared with 1977 and 1987 surveys and reveals several claims to professional autonomy. PMID- 10693094 TI - Ibuprofen use to reduce the incidence and severity of bronchopulmonary dysplasia: a pilot study. AB - OBJECTIVE: To assess the safety and efficacy of ibuprofen in reducing the incidence and severity of bronchopulmonary dysplasia (BPD) in preterm infants. METHODS: A total of 18 premature infants between 23 and 28 weeks' gestation were studied. Ibuprofen (10 mg/kg of loading dose followed by 5 mg/kg every 12 hours) was administered intravenously or orally to nine infants on respiratory support at > or = 7 days of age and was continued until 28 days of life or until the infants were spontaneously breathing room air, whichever occurred earlier. Ibuprofen levels in plasma were measured in five of these infants. The outcome variables (BPD, ventilatory parameters, and complications) in the study group were compared with those in nine matched controls treated conventionally. RESULTS: The incidence of BPD at 36 weeks postconceptional age was similar in both groups (eight of nine in each group). The percentage of oxygen requirement, the ventilatory efficiency index, and steroid use were also similar in both groups. One infant in the study group, who was also receiving steroids and aminophylline, developed gastrointestinal hemorrhage. Reversible renal failure in one infant and necrotizing enterocolitis in another infant were seen at 4 and 21 days, respectively, after the last dose of ibuprofen. There was no difference in the incidence of intraventricular hemorrhage between the two groups. Plasma levels of ibuprofen at 3 hours after the first dose ranged from 10.3 to 36 mg/liter. Study infants tolerated the feeds better and achieved the full enteral goal earlier than controls (p = 0.04). CONCLUSION: Although a trend toward less ventilator and hospital days in the ibuprofen group was observed in this pilot study, the differences were not statistically significant. The incidence of BPD was similar in both groups. In the study group, two infants developed gastrointestinal complications and a third infant experienced reversible renal failure. The plasma ibuprofen levels were low. Further studies are needed to assess the use of ibuprofen for the prevention and/or treatment of BPD in preterm infants. PMID- 10693095 TI - Does supine positioning increase apnea, bradycardia, and desaturation in preterm infants? AB - OBJECTIVE: The purpose of this study was to determine the effects of prone and supine positioning on the cardiorespiratory stability of preterm infants with apnea and bradycardia. METHODS: A total of 22 preterm infants with symptomatic apnea and bradycardia (gestational age of 26.9 +/- 1.8 weeks and birth weight of 865 +/- 235 gm) were monitored for 24 hours (in four sequential 6-hour blocks) for apnea, bradycardia, and oxygen desaturation in alternating positions (prone or supine) following randomization. Postconceptional age at the time of study was 31.9 +/- 3.0 weeks. Respiratory rate, heart rate, and transcutaneous oxygen saturation were continuously monitored. All episodes of apnea (> or = 10 seconds), bradycardia (< 100 beats per minute), and oxygen desaturation (< 90%) were recorded on an event monitor. Episodes of apnea, bradycardia, and oxygen desaturation were defined as clinically significant if the following criteria were met: apnea, > or = 15 seconds; bradycardia, < 90 beats per minute; and oxygen desaturation, < 80%. All other recorded episodes were considered mild. The episodes were analyzed for statistical significance using the paired t-test. RESULTS: No significant differences (p > 0.05) in the incidence of clinically significant apnea, bradycardia, or desaturation between supine and prone positions were seen in these preterm infants. CONCLUSION: Our results suggest that the cardiorespiratory stability of preterm infants is not significantly compromised by supine positioning. PMID- 10693096 TI - Impact of infants born at the threshold of viability on the neonatal mortality rate in Colorado. AB - OBJECTIVE: To determine the contribution of infants born at the threshold of viability (< 750 gm) on neonatal mortality in Colorado. STUDY DESIGN: For the period of January 1991 to December 1996, all Colorado live births who expired were evaluated for gestational age, birth weight, gender, hospital level of care, age at time of death, delivery room resuscitation, mechanical ventilation, medical and surgical complications, and serious malformations. RESULTS: Although infants weighing < 750 gm represent only 0.31% of all live births, they account for 46.3% of deaths. While those infants weighing < 500 gm and with a gestation of < 24 weeks almost always died (94.7%), the majority born in the 500- to 745-gm category (55.8%) survived. The vast majority (88.5%) of deaths occurred on the first day of life. A total of 38.4% of births in which the infant weighed < 750 gm occurred outside bona fide regional perinatal centers. CONCLUSION: Future attempts to reduce the Colorado neonatal mortality rate would best focus on the 500- to 750-gm weight group through the re-regionalization of high-risk perinatal care. PMID- 10693097 TI - Nitroglycerin for rapid tocolysis: development of a protocol and a literature review. AB - OBJECTIVE: A protocol for nitroglycerin (NTG) use based on experiences with regard to new and previously described obstetric cases is presented. The efficacy of NTG tocolysis for obstetric emergencies is clinically evaluated. STUDY DESIGN: Hemodynamically stable parturients requiring acute tocolysis were treated with intravenous NTG and closely monitored. Clinical information was subsequently abstracted from medical records and compared with data from previous reviews. RESULTS: Tocolytic treatment was successful in all cases (22 of 22, 100%). Complications were clinically insignificant. The most common problem was transient hypotension, which occurred in 9 of 22 (41%) cases. CONCLUSION: NTG is an effective tocolytic with minimal complications, rapid onset, and a brief half life. These characteristics favor its use during select obstetric procedures. However, strict adherence to protocols for administration is advised. PMID- 10693098 TI - Postnatal care in low-income urban African American women: relationship to level of prenatal care sought. AB - OBJECTIVE: This study examined the relationship between level of prenatal care utilization and postnatal patterns of health care behavior among high-risk minority women. The primary hypothesis was that prenatal care utilization predicts subsequent levels of both the maternal and child health services used in the postnatal period. METHODS: The study population consisted of 297 low-income African American women who were recruited at delivery at an urban tertiary medical center in the Mid-Atlantic region. They were followed monthly for 1 year using telephone interviews to determine their use of maternal and child health services. Four levels of prenatal care were identified retrospectively based on reviews of health records and screening interviews using the Kessner Index. Data regarding pregnancy outcomes, maternal postnatal visits, and well-child and acute care child visits were collected. RESULTS: Women who sought inadequate or no prenatal care had greater infant morbidity and mortality in the postnatal period and significantly lower levels of attendance at maternal postnatal visits, well child visits, immunization completions, and acute care visits. CONCLUSION: This study confirms that the level of prenatal care is indicative of the level of postnatal care women seek for themselves and their children in the first year after delivery. PMID- 10693099 TI - Pulmonary function and electrolyte balance following spironolactone treatment in preterm infants with chronic lung disease: a double-blind, placebo-controlled, randomized trial. AB - OBJECTIVE: To study the effect of spironolactone on dietary electrolyte supplementation, pulmonary function, and electrolyte balance in premature infants with chronic lung disease. STUDY DESIGN: A double-blind, randomized, and placebo controlled trial was designed to study two groups of low birth weight infants with chronic lung disease at Pennsylvania Hospital. The placebo group received chlorothiazide and a placebo, and the spironolactone group received chlorothiazide and spironolactone during the 2-week study period. A two-tailed t test was used to determine equivalence between the two groups. RESULTS: Pulmonary compliance, resistance and tidal volume, serum sodium and potassium, and FIO2, were not statistically different between the two groups. The need for sodium and/or potassium chloride did not differ between the two groups, nor did the quantity of each salt. CONCLUSION: The addition of spironolactone did not reduce the requirement for supplemental electrolytes, nor did it improve pulmonary mechanics or electrolyte balance. PMID- 10693100 TI - Gastrointestinal function and growth in premature infants: is non-nutritive sucking vital? AB - OBJECTIVE: To determine the importance of non-nutritive sucking (NNS) in the development of gastrointestinal function and growth in premature infants. DATA SOURCES: A systematic computerized search of MEDLINE, the Cumulative Index of Nursing in Allied Health Literature, Health, Best Evidence, and the Cochrane Library was performed. STUDY SELECTION: The search yielded eight randomized controlled studies relative to the outcomes of interest: sucking response, gastric emptying, weight gain, and time to discharge from hospital. DATA EXTRACTION: Relevant articles were selected using published criteria for detecting clinically sound studies and evidence-based information. DATA SYNTHESIS: NNS reduces length of hospitalization; however, its effect on the other variables was inconclusive. CONCLUSION: There is a lack of agreement concerning the outcomes of interest, apart from the positive contribution of early hospital discharge. The studies were methodologically flawed, which compromised validity and estimation of the treatment effect. NNS cannot be currently recommended as a beneficial intervention. PMID- 10693101 TI - Fatal neonatal Salmonella enteritidis sepsis. AB - A case history of a fatal neonatal infection caused by Salmonella enteritidis group D is reported. The baby deteriorated rapidly at 24 hours after birth with clinical signs and symptoms of an acute abdomen. Bloody diarrhea led to a tentative diagnosis of midgut volvulus or necrotizing enterocolitis. Autopsy and bacteriologic investigation revealed sepsis by S. enteritidis group D. The same organism was found in cultures taken from stool and vaginal swabs from the mother. This clearly confirmed transmission of the infection during delivery. PMID- 10693102 TI - Acute suppurative parotitis: uncommon presentation in a premature infant. PMID- 10693103 TI - Subdiaphragmatic pulmonary sequestration: a case report with review of the literature. PMID- 10693104 TI - Placental pathology casebook. Serpentine aneurysms of the placenta with fetal consequences. AB - Two cases of placental surface vessel aneurysms are presented. One case was associated with severe fetal intrauterine growth restriction and fetal thrombocytopenia. The other case was associated with "molar transformation" of placental villous tissue. PMID- 10693105 TI - Special imaging casebook. High imperforate anus with enterolithiasis and dysraphism with large meningomyelocele. PMID- 10693106 TI - Fetal heart rate monitoring casebook. Amnioinfusion as fetal therapy. AB - Amniotic fluid (AF) plays multiple roles in fetal development and wellbeing. A global consideration of the possibilities of AF manipulation allows for the maximum benefits to be derived from assessing and selectively augmenting AF in clinical practice. PMID- 10693107 TI - Mental health: a report of the Surgeon General. PMID- 10693108 TI - Abortion: a psychiatric patient's right to choose. AB - 1. Competent mentally ill patients should be allowed to terminate pregnancy. 2. It is the responsibility of the staff to provide the patient with a safe, supportive, and respectful environment. 3. Early and frequent staff processing opportunities should be offered to clarify any underlying issues. PMID- 10693109 TI - Use of physical restraints on hospitalized psychogeriatric patients. AB - 1. When making decisions about the use of physical restraints, minimizing the physical risk of patients was determined to be the most important obligation. 2. Nurses' understanding and consideration of important factors that influence restraint use are essential to its reduction. 3. Nurses should be aware of their ethical responsibilities to older patients and their families. They must ensure the patient's right to be informed and respect their dignity. PMID- 10693110 TI - Nurses' burnout: an existential psychodynamic perspective. AB - 1. According to Psychodynamic Theory the reason why most people choose a career has to do with childhood experiences. 2. Both qualitative and quantitative data suggest that nurses' burnout is caused by a failure to derive existential significance from work. 3. Nurses' sense of significance seems to be related to control and is often caused by childhood experiences of lack of control. PMID- 10693111 TI - Resolving alcohol problems using an online self-help approach: moderation management. AB - As illustrated by the emergent MM approach discussed in this article, a paradigm shift is occurring in relation to how alcohol problems are defined and managed. Moreover, one of the most interesting aspects of this transformation is the way in which similar belief systems are simultaneously emerging from divergent lay and professional groups. In spite of this flurry of activity and optimism, however, nurses should not embrace new frameworks and treatment strategies. Rather, the charge to our discipline and others is to think critically, research the effectiveness of newly proposed treatment approaches, and recommend and implement promising strategies with discernment. PMID- 10693112 TI - A nurse's guide to mental health assessment reference sources. AB - 1. As the research base on psychological testing has proliferated across various disciplines, a comprehensive review of the testing literature is recommended. 2. In researching literature on psychological tests, nurses need to rely on traditional print as well as electronic databases and Internet sites. 3. In reviewing test items or in using a test, nurses need to be aware of test-user qualifications and publisher copyright restrictions. PMID- 10693113 TI - Interaction effects of emotional status and sexual abuse on adult sexuality. AB - This study examined the additive, interaction effects of emotional status with sexual abuse on adult sexual functioning and sexual responsibility. The Golombok Rust Inventory of Sexual Satisfaction (GRISS; Rust & Golombok, 1986), the Personality Assessment Inventory (PAI; Morey, 1991), and a questionnaire regarding sexual experiences, number of unwanted pregnancies, number of unsafe sexual partners, and sexual abuse history, were administered to 200 psychology students. One hundred and forty-three participants were retained in the study. Two-way multivariate analyses of variance (MANOVA) were conducted for the sexual functioning variables (as measured by the GRISS), while two-way analyses of variance (ANOVA) were conducted for the sexual irresponsibility variables (as measured by the sexual experiences questionnaire). It was found that women who have high anxiety scores on the PAI and have a history of sexual abuse reported higher numbers of unwanted pregnancies, while sexual abuse history was not associated with numbers of unwanted pregnancies for women with lower levels of anxiety. Results were not significant, however, for the sexual functioning variables. In addition, depression and alcoholism did not have interacting effect on the association between sexual abuse history and any of the sexuality variables. These results may suggest that the effects of sexual abuse on adult sexuality may not be as pervasive as was once thought. PMID- 10693114 TI - The Arizona Sexual Experience Scale (ASEX): reliability and validity. AB - Although sexual dysfunction is common in psychiatric patients, quantification of sexual dysfunction is limited by the paucity of validated, user-friendly scales. In order to address this problem, the authors have developed the Arizona Sexual Experiences Scale (ASEX), a five-item rating scale that quantifies sex drive, arousal, vaginal lubrication/penile erection, ability to reach orgasm, and satisfaction from orgasm. Possible total scores range from 5 to 30, with the higher scores indicating more sexual dysfunction. This study assesses the internal consistency, test-retest reliability, and convergent and discriminant validity of the ASEX. PMID- 10693115 TI - A study using Viagra in a mental health practice. AB - The clinical responses of 58 men who were given Viagra between April and December 1998 in a psychiatric outpatient setting were tracked through 2 follow-up visits. Four categories of success and 3 categories of failure are defined to describe the outcomes. Although 43% had ideal outcomes--cure or uncomplicated drug dependent successes--50% again were able to have intercourse and 63% experienced improved erections. Seventeen percent of the original sample did not benefit from the drug; 21% were lost to follow-up. This intent-to-treat study adds more evidence of Viagra's effectiveness and further clarifies the need to focus on psychodynamic considerations when trying to assist the majority of patients who did not have ideal outcomes. PMID- 10693116 TI - The female sexual response: a different model. AB - Clarification of women's sexual response during long-term relationships is needed. I have presented a model that more accurately depicts the responsive component of women's desire and the underlying motivational forces that trigger it. The variety of arousal/orgasm responses is also acknowledged. The purpose is both to prevent diagnosing dysfunction when the response is simply different from the traditional human sex-response cycle and to more clearly define subgroups of dysfunction. The latter would appear to be necessary before progress in newer treatment modalities, including pharmacological, can be made. PMID- 10693117 TI - Cognitive distraction and women's sexual functioning. AB - Past research on the role of cognitive distraction in sexual dysfunction typically has focused on males and has been conducted in the laboratory using artificial stimuli. In the current study, young adult women (N = 74) with coital experience completed questionnaires regarding cognitive distraction and their sexuality. Those women who reported greater cognitive distraction during sexual activity with a partner also reported relatively lower sexual esteem, less sexual satisfaction, less consistent orgasms, and higher incidence of pretending orgasm even after the women's general affect, sexual desire, general self-focus, general sexual attitudes, and body dissatisfaction were statistically controlled. Results are discussed with regard to directions for future research and implications for sex therapy. PMID- 10693118 TI - Female analogies to perversion. AB - Unlike the intrapsychic mechanism for self-esteem regulation in males as a basic component of perversion--extrapsychically (compensationally) culminating in an output of sexual impulses--a functional stabilization of the female self-concept seems more likely if conflict drives were to be focused on reproductional aspects and not on sexuality. It therefore seems more suitable to use a new expression in linguistic analogy to perversion: "reproversion." The case history gives an example of a clinical manifestation of "reproverse" symptom formation. The general survey describes the main points of view in regard to clinically oriented differentiation, i.e., intensity, ego-proximity in the personality structure, and one's own self-acceptance within "reproverse" symptom formation. Underlying personality disturbances are also discussed. The significance of reproversion is relevant to many different specialized medical fields. This is explained in conclusion, using the examples of denied pregnancy and infanticide at birth based on initial empirical results. PMID- 10693119 TI - College students' reasons for nonuse of condoms within dating relationships. AB - Two hundred ten heterosexual undergraduates in dating relationships were surveyed about reasons for not using condoms every time for vaginal and anal sex and for increasing or decreasing condom use during their relationships. Half of the respondents reported consistent condom use in the first month of their relationships, while only 34% reported consistent condom use in the past month. Subjective assessments of partner safety and the belief that sufficient measures were being taken to avoid pregnancy were important reasons for condom nonuse. Study results suggest that interventions should emphasize the importance of objectively assessing HIV/STD risk before reducing condom use within relationships. Interventions also need to provide additional information on the riskiness of heterosexual anal sex. PMID- 10693120 TI - [Models of radiation dispersion in medical laser tomographic system]. AB - The prospects for developing a new noninvasive medical diagnostic technique, such as optical (laser) tomography of biological tissues largely depend on the feasibility of designing an adequate mathematical model for the reconstruction of images of the spatial structure of objects that are in the highly scattering medium (HSM). The basic difficulty is that it is necessary to effectively describe the physical properties and the propagation pattern of photons of different types in such a medium. This review considers methods for describing the distribution of laser ultrashort pulses in HSM with particular emphasis on a new nonstationary two-flux model of radiation transfer. PMID- 10693121 TI - [Possibilities of localization of the areas of myocardial depolarization impairment using ECG high-frequency filtration]. AB - The paper deals with the physiological processes of myocardial depolarization and repolarization and the way of solving the problems in the three-dimensional plotting the disorders occur. It reviews basic papers on this problem. Attempts are made to explain the occurrence of recorded late myocardial potentials and the technical ways of identifying the areas of impaired myocardial depolarization are proposed. This approach may be considered to be offers for designing new devices and computer programmes having promising capacities to visualize ECG. PMID- 10693123 TI - [Organizing diagnosis of parameters of limb loading during transosseous osteosynthesis by external fixation devices]. AB - The presented system for compression-traction osteosynthesis (CTS) involves a modified Ilizarov apparatus, a set of force and length detectors, PC controlled measuring system, a set of cables and original software. The system apparatus allows for angular displacement of bone fragments in the range of +/- 30 degrees or 0-60 degrees and lateral displacement of bone fragment edges in the joint along any azimuth in the ring plane. It was mathematically proved that the axial and angular displacement of the two apparatus ring blocks relative to each other can be described with three parameters: axial displacement of the ring center, the ring deflection (turn) angle and the ring axis turn azimuth (angle). Formulas for calculation of the displacement of three pairs of displacement control nuts so the plane of the fracture displacement angle (correction) cross the axis of the required ring plane turn azimuth (angle) were deduced for the reciprocal angular displacement of the bone fractures. No additional removable joints or the apparatus reconstruction for angular fracture displacement are needed. The CTS features the display of the current reciprocal position of bone fragment axes during the course of apparatus loading on the PC monitor. PMID- 10693122 TI - [Comparative effectiveness of the use of hematological methods to analyze the procoagulant activity of biological materials]. AB - Whether clinical hematological methods by employing two parameters: (plasma recalcification time (PRT) and activated partial thromboplastin time (ACTT) can be used to examine the procoagulant activity of the surface of biological materials was comparatively studied. The method PRT was shown to be sensitive and reproducible while ACTT failed to significantly record differences in the procoagulant activities of various materials. The authors chose the optimal protocol for evaluating the blood compatibility of biological materials: the surface area/plasma volume ratio is more than 150 cm2/ml, the incubation time of samples is no less than 180 sec at 37 degrees C and continuous mixing. PMID- 10693124 TI - [EEG-Expert automated diagnostic system]. AB - The system is designed to describe and collect data of virtual analysis of electroencephalograms (EEG), as well as to make an expert conclusion on the functional status of the brain and its particular regions. The system includes three parallelly operating systems in the form of a EEC visual analysis schedule as a questionnaire. A programme provides recommendations for describing the level, pattern, and magnitude of the changes detected. Another programme automatically makes an EEC description and a conclusion on cerebral function in conformity with the signs given in the questionnaire and displays them on the screen and sends them to the print. The system may be used for practice in the in and outpatient settings, for training and postgraduate training of young specialists, and for researches. PMID- 10693125 TI - [New technological means in medical diagnosis]. AB - Diagnostic methods using a Pulsar radar measuring computer unit have been developed. The methods are based on the contactless recording of respiratory parameters and pulse and they evaluate human body's functions in real time from the measurements of the duration of cardiac rhythms and calculate the differential function of their distribution and the spectrum of a cardiac rhythmogram. PMID- 10693126 TI - [Mathematical processing of human platelet distribution according to size for determination of cell heterogeneity]. AB - The paper proposes a method for mathematical treatment of the distribution of human platelets by sizes to detect the heterogeneity of cell populations. Its use allowed the authors to identify three platelet populations that have different parameters of size distribution. The proposed method opens additional vistas for analyzing the heterogeneity of platelet populations without sophisticating experimental techniques. PMID- 10693127 TI - [Microwave technology in dental and eye prosthesis]. AB - The paper outlines the specific features of using microwave technologies in dental and eye prostheses designed in Russia in 1992-1998. Thermal, microwave, and combined polymerizations of plastic materials used in prosthesis are compared. A Denta microwave unit is presented, which realizes a combined microwave technology. PMID- 10693128 TI - [Use of impedance measurements of the permeability of skin capillaries to estimate the activity of tuberculous process]. AB - The paper considers whether a new impedance measurement technique can be used to estimate the activity of the process. Following 6 hours of intradermal tuberculin administration, the permeability of capillaries proved to be of the maximum informative value, which provides not only a judgement of the tuberculous origin of the disease, but evaluates the activity of the process, by allowing for the inclusion of this technique into a package of diagnostic measures in patients having the suspected origin of the disease. PMID- 10693129 TI - Manganese in natural waters and earth's crust: its availability to organisms. PMID- 10693130 TI - Manganese transport in microorganisms. PMID- 10693131 TI - Manganese uptake and transport in plants. PMID- 10693132 TI - Manganese metabolism in animals and humans including the toxicity of manganese. PMID- 10693133 TI - Interrelations between manganese and other metal ions in health and disease. PMID- 10693134 TI - The use of manganese as a probe for elucidating the role of magnesium ions in ribozymes. PMID- 10693135 TI - Mn2+ as a probe of divalent metal ion binding and function in enzymes and other proteins. PMID- 10693136 TI - Enzymes and proteins containing manganese: an overview. PMID- 10693137 TI - Manganese(II) in concanavalin A and other lectin proteins. PMID- 10693138 TI - Manganese-activated phosphatases. PMID- 10693139 TI - Manganese(II) as a probe for the mechanism and specificity of restriction endonucleases. PMID- 10693140 TI - Role of the binuclear manganese(II) site in xylose isomerase. PMID- 10693141 TI - Arginase: a binuclear manganese metalloenzyme. PMID- 10693142 TI - The use of model complexes to elucidate the structure and function of manganese redox enzymes. PMID- 10693143 TI - Manganese(II)-dependent extradiol-cleaving catechol dioxygenases. PMID- 10693144 TI - Manganese catalases. PMID- 10693145 TI - Manganese peroxidase. PMID- 10693146 TI - Manganese superoxide dismutase. PMID- 10693147 TI - Mechanistic aspects of the tyrosyl radical-manganese complex in photosynthetic water oxidation. PMID- 10693148 TI - The polypeptides of photosystem II and their influence on manganotyrosyl-based oxygen evolution. PMID- 10693149 TI - The effects of early rearing environment on the development of GABAA and central benzodiazepine receptor levels and novelty-induced fearfulness in the rat. AB - We compared the effects of handling or maternal separation from the day following birth until postnatal day 14 on behavioral responses to novelty and on GABAA and central benzodiazepine (CBZ) receptor levels in the rat. As adults, handled animals showed reduced startle responsivity, increased exploration in a novel open field, and decreased novelty-induced suppression of feeding relative to the handled (H) and/or maternal separation (MS) groups. As compared with handled animals, both nonhandled (NH) and MS animals displayed: (1) reduced GABAA receptor levels in the locus coeruleus (LC) and the n. tractus solitarius (NTS); (2) reduced CBZ receptor sites in the central and lateral n. of the amygdala, the frontal cortex, and in the LC and NTS; and (3) reduced levels of the mRNA for the gamma 2 subunit of the GABAA receptor complex, which confers high affinity BZ binding, in the amygdaloid nuclei as well as in the LC and NTS. Both the amygdala and the ascending noradrenergic systems have been considered as critical sites for the anxiolytic effects of benzodiazepines. These data suggest that early life events influence the development of the GABAA receptor system, thus altering the expression of fearfulness in adulthood. PMID- 10693150 TI - Behavioral effects of central administration of the novel CRF antagonist astressin in rats. AB - Astressin, a novel corticotropin releasing factor (CRF) antagonist, has been found to be particularly potent at inhibiting the hypothalamo-pituitary-adrenal axis. The aim of the present study was to determine the effects in rats of astressin in attenuating the anxiogenic-like response produced by social stress and intracerebroventricular (ICV) CRF administration on the elevated plus-maze, and ICV CRF-induced locomotor activation in the rat. Astressin significantly reversed the anxiogenic-like response induced by both social stress and ICV rat/humanCRF (r/hCRF) on the elevated plus-maze, but failed to block the effects of r/hCRF-induced locomotor activity in a familiar environment. When these results were compared to previous studies performed with the same paradigms using other CRF antagonists, astressin showed effects similar to those of D-PheCRF(12 41) on plus-maze performance. However, contrary to alpha-helicalCRF(9-41) and D PheCRF(12-41), astressin had no effect on CRF-induced locomotor activity. These results suggest that astressin may have a unique anti-CRF profile compared to previously tested antagonists. PMID- 10693151 TI - Seasonal variation in CSF 5-HIAA concentrations in male rhesus macaques. AB - Seasonal changes in cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid (5-HIAA) concentrations were assessed on multiple occasions in 103 free-ranging male rhesus macaques (Macaca mulatta). At the time of sampling subjects ranged between the ages of 2 and 6 years. CSF samples were collected between the hours of 0900 and 1600 throughout the Fall, Winter, and Spring from 1990 through 1994. Data were analyzed in a general linear mixed model with random intercepts. Results indicated that CSF 5-HIAA concentrations decreased with age. CSF 5-HIAA concentrations were significantly increased in the Fall (October and November), which is the height of the breeding season, with no evidence of differences between Winter and Spring. There was also some evidence that the seasonal variation in CSF 5-HIAA concentrations was blunted in younger, more immature subjects. PMID- 10693152 TI - Increased ACTH concentrations associated with cholecystokinin tetrapeptide induced panic attacks in patients with panic disorder. AB - Preclinical findings on the role of corticotropin releasing hormone (CRH) in stress and anxiety, on the interaction of CRH and cholecystokinin (CCK) in modulating anxiety, as well as the blunted corticotropin (ACTH) response to CRH in panic disorder suggest that CRH may play a role in panic disorder. To further characterize the role of the hypothalamic-pituitary-adrenocortical (HPA) system in panic disorder, we compared patients with and without CCK tetrapeptide (CCK-4) induced panic attacks. Twenty-four patients with panic disorder were given injections of CCK-4 (25 micrograms). Panic attacks, psychopathological changes, as well as ACTH and cortisol secretion were recorded. Fifteen of the 24 patients experienced a panic attack after CCK-4. ACTH secretion was significantly higher in the patients with CCK-4-induced panic attacks than in those without such attacks. The patients without CCK-4-induced attacks had a brief but less pronounced increase in ACTH concentrations. Cortisol concentrations were not significantly increased after CCK-4 administration. The increased ACTH concentrations suggest that the activation of the HPA system in CCK-4-induced panic attacks plays a physiological role. CRH may be involved in experimentally occurring and perhaps in naturally-occurring panic attacks as well. PMID- 10693154 TI - Atrophy and high intensity lesions: complementary neurobiological mechanisms in late-life major depression. AB - The primary objective of our study was to examine the role of atrophy, high intensity lesions and medical comorbidity in the pathophysiology of major depressive disorder in the elderly (late-life MDD). Our sample was comprised of 51 patients with late-life MDD and 30 non-depressed controls. All subjects were scanned on 1.5 tesla magnetic resonance imaging scanner (MRI) and absolute and normalized measures of brain and lesion volumes were obtained and used for comparison between groups. Patients with MDD had significantly smaller frontal lobe volumes, together with larger whole brain lesion volumes when compared with controls (p < .05). Whole brain lesion volumes correlated significantly (r = 0.41, p = .006) with overall medical comorbidity. The odds ratio (OR) for existing MDD increases significantly with a decrease in frontal lobe volume and an increase in whole brain lesion volumes (p < .05). Our findings suggest that atrophy and high intensity lesions represent relatively independent pathways to late-life MDD. While medical disorders lead to neuropathological changes that are captured on MR imaging as high intensity signals, atrophy may represent a relatively autonomous phenomenon. These findings have broad implications for the pathophysiology of mood disorders and suggest that complementary neurobiological processes may lead to cumulative neuronal injury thereby predisposing to clinical depression. PMID- 10693153 TI - Serotonin function following remission from bulimia nervosa. AB - Abnormal serotonergic regulation in bulimia nervosa is thought to contribute to recurrent binge eating, depressed mood, and impulsivity. To follow-up on previous studies showing decreased neuroendocrine responses in symptomatic patients, this study assessed serotonin-mediated prolactin responses in individuals who had remitted from bulimia nervosa. Subjects included 21 women with a history of bulimia nervosa and 21 healthy female controls, as well as an additional comparison group of 19 women with current bulimia nervosa. Placebo-controlled neuroendocrine response studies utilized a single oral dose (60 mg) of the indirect serotonin agonist d,l-fenfluramine. For the bulimia nervosa remitted group, the fenfluramine-stimulated elevation in serum prolactin concentration was not significantly different from the response in healthy controls, but was significantly larger than the response in patients with current bulimia nervosa (p < .01). These findings suggest that diminished serotonergic neuroendocrine responsiveness in bulimia nervosa reflects a state-related abnormality. The results are discussed in relationship to recent reports indicating that some alterations in central nervous system serotonin regulation may persist in symptomatically recovered individuals. PMID- 10693155 TI - Peripheral markers of serotonergic and noradrenergic function in post-pubertal, caucasian males with autistic disorder. AB - Some studies have suggested that disorders in the peripheral and central metabolism of serotonin (5-HT) and noradrenaline may play a role in the pathophysiology of autistic disorder. This study examines serotonergic and noradrenergic markers in a study group of 13 male, post-pubertal, caucasian autistic patients (age 12-18 y; I.Q. > 55) and 13 matched volunteers. [3H] paroxetine binding Kd values were significantly higher in patients with autism than in healthy volunteers. Plasma concentrations of tryptophan, the precursor of 5-HT, were significantly lower in autistic patients than in healthy volunteers. There were no significant differences between autistic and normal children in the serum concentrations of 5-HT, or the 24-hr urinary excretion of 5-hydroxy indoleacetic acid (5-HIAA), adrenaline, noradrenaline, and dopamine. There were no significant differences in [3H]-rauwolscine binding Bmax or Kd values, or in the serum concentrations of tyrosine, the precursor of noradrenaline, between both study groups. There were highly significant positive correlations between age and 24-hr urinary excretion of 5-HIAA and serum tryptophan. The results suggest that: 1) serotonergic disturbances, such as defects in the 5-HT transporter system and lowered plasma tryptophan, may play a role in the pathophysiology of autism; 2) autism is not associated with alterations in the noradrenergic system; and 3) the metabolism of serotonin in humans undergoes significant changes between the ages of 12 and 18 years. PMID- 10693156 TI - Putamen mitochondrial energy metabolism is highly correlated to emotional and intellectual impairment in schizophrenics. AB - In a recent study, we demonstrated that cytochrome-c oxidase (COX), an indicator of neuronal activity, is increased in several brain regions from chronic, medicated schizophrenics. In the present study, to address the functional significance of those findings, we have measured COX activity in a group of schizophrenics in whom antemortem geriatric measures of motor, intellectual, and emotional impairment had been assessed. COX activity in the putamen was strongly negatively correlated with emotional (r = -.76; p < .005) and intellectual impairment (r = -0.76; p < .005), but not with motor impairment (r = 0.01). No significant correlations could be found in the frontal cortex, thalamus, caudate nucleus, globus pallidus, mesencephalon, or nucleus accumbens. Dopamine D2 receptor density in the putamen, measured with [3H]raclopride, was elevated in schizophrenics as compared to controls, as were Kd values. In contrast to COX activity, D2 receptor binding was moderately, but significantly positively correlated with intellectual impairment (r = 0.64; p < .05) but not with motor impairment. Results expose a unique anomaly in the effects of neuroleptics in terms of increasing neuronal signaling in the putamen, which may underlie a reversal of cognitive deficits in schizophrenics, while at the same time, elevating D2 receptor density that seems to be detrimental. PMID- 10693157 TI - The effects of a sub-anaesthetic dose of ketamine on human selective attention. AB - A growing number of studies demonstrate that antagonists of the N-methyl-D aspartate (NMDA) receptors can induce a broad range of psychophysiological anomalies in healthy subjects similar to those observed in schizophrenia. In this study, the effect of a sub-anaesthetic dose of the non-competitive NMDA antagonist, ketamine, on human selective attention was explored. It was hypothesized that ketamine would induce in healthy subjects psychophysiological anomalies that are commonly observed in schizophrenic patients, such as reduced P300 amplitude and a reduction of both mismatch negativity (MMN) and processing negativity (PN). In a double-blind randomized placebo-controlled design, healthy male volunteers (n = 18) were challenged with a sub-anaesthetic dose of ketamine (0.3 mg/kg i.v.) after which they were tested in a selective attention task. In this task, two types of stimuli were evenly presented to the left or right ear: standard tones (80%) and deviant tones (20%) of either 1000 or 1100 Hz. The duration of a stimulus (95 dB) was 50 ms, the interstimulus intervals were randomized between 1750 and 2150 ms. The volunteer was instructed to push a button as quickly as possible after hearing the deviant tone in a specified ear. Ketamine did not alter performance of the subjects: in both the placebo and drug condition their reaction times for and percentages of hits and false alarms did not differ. Ketamine did, however, reduce PN and the P300 amplitude (both in general and to deviant stimuli in particular). However, no drug effect on MMN was found. In addition, ketamine enhanced the N100 amplitude to deviant stimuli. In conclusion, ketamine induces some of the attentional deficits in healthy controls that are observed in schizophrenic patients. Consequently, reduced glutamatergic activity in the brain may be involved in some of the symptoms of schizophrenia. PMID- 10693158 TI - Assessing the efficacy of treatments for the deficit syndrome of schizophrenia. AB - The primary, enduring negative symptoms found in some patients with schizophrenia have become the focus of clinical treatment trials, but there has been no consensus on the best methods for approaching this area. In future trials, a number of issues need to be considered, including analytic strategies, the limitations in instruments used to measure negative symptoms, and study design. An appropriate design for establishing the efficacy of treatments for the deficit syndrome is proposed. PMID- 10693159 TI - Effects of atypical neuroleptics on sustained attention deficits in schizophrenia: a trial of risperidone versus haloperidol. AB - To help determine whether sustained attention deficits as measured with the Continuous Performance Test (CPT) are stable vulnerability indicators of schizophrenia, we compared the CPT performance of schizophrenic patients before and after treatment with risperidone or haloperidol. In this double blind trial, 56 schizophrenic patients were randomly assigned to a 12-week regimen of either risperidone or haloperidol, after a 1-week washout period. The patients undertook two sessions of the CPT (undegraded and 25% degraded) twice, one at the end of the washout period and the other at the end of the study. Thirty-eight patients completed the study, 19 in each group. Both groups experienced significant improvements in clinical symptoms, and the risperidone group showed no change in the severity of extrapyramidal symptoms. Despite those improvements, the CPT performance indexes did not change significantly from the beginning to the end of the study. These findings indicate that sustained attention deficits might be stable vulnerability indicators of schizophrenia. PMID- 10693160 TI - Adrenergic and noradrenergic plasma levels in Lesch-Nyhan disease. AB - Noradrenergic dysfunction and abnormality in monoamine oxidase (MAO) enzyme activity have been reported previously in Lesch-Nyhan (LN) disease. This study examines peripheral indices of adrenergic, noradrenergic, and MAO function in children and young adults with LN disease (n = 11), and healthy subjects (n = 9). Blood samples, collected in identical conditions prior to a positron emission tomography (PET) study, were assayed for concentrations of epinephrine (EPI), norepinephrine (NE), and 3-methoxy-4-hydroxyphenylglycol (DHPG) (which results from the degradation of NE by monoamine oxidase type A [MAO-A]). The LN subjects had significantly higher EPI levels by 245% (p < .00) and lower DHPG levels by 42% (p < .00) compared to the control group. No group differences were noted in NE plasma levels. Cognitive function (IQ tested by Stanford Binet Intelligence Scale) was associated with EPI in the LN group (r = 0.77, p = .009), but not in the control group. The abnormally high EPI plasma concentrations may indicate another biochemical dysfunction secondary to the absence of the HPRT enzyme in LN patients. Such a biochemical deficit is likely to originate from the adrenal medulla, which is the primary site of EPI synthesis. The adrenal medulla may be directly affected by the absence of hypoxanthine guanine phosphoribosyl transferase (HPRT) enzyme, or may receive inappropriately high descending activation input from the brain. The abnormally low DHPG levels, in the context of normal NE levels, indicates low MAO activity, either as a primary deficit, or as secondary adaptive changes to spare NE levels that would otherwise be too low for adequate noradrenergic function. PMID- 10693161 TI - Increased temporal cortex ER stress proteins in depressed subjects who died by suicide. AB - Regulation of ER stress proteins, such as the 78-kilodalton glucose regulated protein (GRP78) by chronic treatment with mood stabilizing drugs suggests that this family of proteins may be involved in the pathophysiology of mood disorders. Indeed, increased levels of GRP78, GRP94, and calreticulin, a third member of the ER stress protein family, were found in temporal cortex of subjects with major depressive disorder who died by suicide compared with controls and subjects who died by other means. No such differences were found in subjects with other psychiatric disorders such as bipolar disorder or schizophrenia. These data suggest a potential role for ER stress proteins in severe depression that merits further study. PMID- 10693162 TI - Changes in nutritional, functional, and inflammatory markers in advanced pancreatic cancer. AB - Weight loss and the acute-phase response have been associated with poor quality of life and survival in advanced pancreatic cancer; however, little information is available on changes in these factors over time and their interrelationships. This study examined changes in weight, Karnofsky performance status, C-reactive protein (CRP), and serum albumin in 25 patients with advanced pancreatic cancer given supportive symptomatic treatment only. Patients were assessed at approximately monthly intervals on a total of 70 occasions, allowing assessment of changes over 37 intervals. Overall, patients had a median weight loss of 2.3 kg/28 days. Median CRP levels rose by 15 mg/l, and serum albumin fell by 1 g/l on average over 28 days. Karnofsky performance status fell by 4 points every 28 days. The nine patients assessed close to death were compared with the 13 assessed close to diagnosis. The increase in CRP level and fall in weight and performance status were significantly greater within eight weeks of death than within eight weeks of diagnosis. Among patients close to diagnosis, 13% had an elevated CRP compared with 100% of those close to death. In multivariate analysis, Karnofsky performance status was significantly associated with percent weight loss and CRP levels. These data further implicate the acute-phase protein response as being associated with the progressive weight loss seen in patients with advanced pancreatic cancer. Changes appear to accelerate close to death. Moreover, the development of cachexia is associated with a reduction in patients' functional capacity. PMID- 10693163 TI - Vegetables, fruits, and related nutrients and risk of breast cancer: a case control study in Uruguay. AB - To evaluate whether the protective effect associated with vegetables and fruits in breast cancer could be explained by nutrients and bioactive substances present in these plant foods, we carried out a case-control study in Uruguay including 400 cases and 405 controls. The intake of vegetables, fruits, and related nutrients was estimated with a food frequency questionnaire on 64 food items. This questionnaire allowed the calculation of total energy intake, and nutrients were calorie adjusted by the residuals method. Odds ratios for study variables were estimated by unconditional multiple logistic regression. Total vegetable, total fruit, dietary fiber, vitamin C, vitamin E, lycopene, folate, and total phytosterol intakes were inversely associated with breast cancer risk [4th quartile odds ratio for total vegetable intake = 0.41, 95% confidence interval = 0.26-0.65, p (for trend) = 0.004]. The association with total vegetable intake was not independent of lycopene intake. The results related to vegetable and nutrient intakes are consistent with antioxidant and antiestrogenic effects. This could be mediated, among other nutrients, by dietary fiber and lycopene intake. The role of other unmeasured phytochemicals, correlated with dietary fiber and lycopene intakes, cannot be ruled out. PMID- 10693164 TI - Effects of long-term treatment of colon adenocarcinoma with crocin, a carotenoid from saffron (Crocus sativus L.): an experimental study in the rat. AB - We used an experimental model in the rat to examine the effects of long-term treatment with crocin, a glycosylated carotenoid from the stigmas of the saffron crocus, on colon cancer. BD-IX rats were divided into four groups: Groups G1 and G2, designated "cancer groups," were used to study the effects of crocin on the progression of colon cancer, and Groups G3 and G4, designated "toxicity groups," were used to study the effects of the treatment on metabolic processes and the parenchyma. DHD/K12-PROb cells were injected subcutaneously into the chest of Group G1 and G2 animals. From 1 to 13 weeks after inoculation, animals in Groups G2 and G4 received a weekly injection of crocin (400 mg/kg body wt s.c.). Animals in Groups G1 and G3 received no treatment. In addition, lines of animal and human colon adenocarcinoma cells (DHD/K12-PROb and HT-29) were used to perform assays in vitro to examine the cytotoxicity of crocin. Life span was extended and tumor growth was slower in crocin-treated female rats, but no significant antitumor effect was found in male rats. Acute tubular necrosis was found in all kidney samples from crocin-treated animals, but slight signs of nephrotoxicity were found by biochemical analysis of the serum. In assays in vitro, crocin had a potent cytotoxic effect on human and animal adenocarcinoma cells (HT-29 and DHD/K12-PROb cells, 50% lethal dose = 0.4 and 1.0 mM, respectively). Treated cells exhibited a remarkable loss of cytoplasm and wide cytoplasmic vacuole-like areas. In conclusion, long-term treatment with crocin enhances survival selectively in female rats with colon cancer without major toxic effects. The effects of crocin might be related to its strong cytotoxic effect on cultured tumor cells. PMID- 10693165 TI - Longitudinal study of weight, appetite, performance status, and inflammation in advanced gastrointestinal cancer. AB - There is increasing evidence that, in most patients with advanced cancer, weight loss is associated with an inflammatory response. To examine the temporal relationship between weight loss, appetite, performance status, and the inflammatory response, 50 patients with advanced gastrointestinal cancer with weight loss were observed for six weeks. Patients were grouped according to whether they had lost weight (> 3%, n = 16), were weight stable (< 3% change, n = 25), or gained weight (> 3%, n = 9). At baseline, the group that subsequently lost weight had lower albumin and higher C-reactive protein concentrations (p < 0.05). On follow-up, there was an increase in C-reactive protein concentration and reductions in triceps skinfold thickness and Karnofsky performance status in the weight-losing group (p < 0.05). In contrast, Karnofsky performance status was improved in the group that gained weight (p < 0.05). Over the six to eight weeks, there was a difference in the changes of triceps skinfold thickness (p < 0.05) and Karnofsky performance status (p < 0.01) between the two groups. These results suggest that loss or gain of > 2.5 kg over a six- to eight-week period is required to produce a significant alteration in performance status in weight losing patients with gastrointestinal cancer. Moreover, the results suggest that the presence of an inflammatory response is associated with further weight loss and the deterioration of performance status. PMID- 10693166 TI - Decreased growth of human prostate LNCaP tumors in SCID mice fed a low-fat, soy protein diet with isoflavones. AB - Epidemiological studies suggest that high intake of dietary fat is a risk factor for the development of clinical prostate cancer. Soy protein has also been proposed to play a role in the prevention of prostate cancer, and one of the isoflavones in soy protein, genistein, inhibits the growth of human prostate cancer cell lines in vitro. This study was designed to evaluate whether altering dietary fat, soy protein, and isoflavone content affects the growth rate of a human androgen-sensitive prostate cancer cell line (LNCaP) grown in severe combined immunodeficient (SCID) mice. SCID mice were randomized into four dietary groups: high-fat (42.0 kcal%) + casein, high-fat (42.0 kcal%) + soy protein + isoflavone extract, low-fat (12.0 kcal%) + casein, and low-fat (12.0 kcal%) + soy protein + isoflavone extract. After two weeks on these diets, the mice were injected subcutaneously with 1 x 10(5) LNCaP tumor cells and placed in separate cages (1 mouse/cage) to strictly control caloric intake. Isocaloric diets were given 3 days/wk, and tumor sizes were measured once per week. The tumor growth rates were slightly reduced in the group that received the low-fat + soy protein + isoflavone extract diet compared with the other groups combined (p < 0.05). In addition, the final tumor weights were reduced by 15% in the group that received the low-fat + soy protein + isoflavone extract diet compared with the other groups combined (p < 0.05). In this xenograft model for prostate cancer, there were statistically significant effects on tumor growth rate and final tumor weight attributable to a low-fat + soy protein + isoflavone extract diet. PMID- 10693167 TI - Colon cancer is induced by a single low dose of azoxymethane in fasted-refed rats. AB - We reported previously that fasting-refeeding enhanced the growth of preneoplastic lesions in the colon of rats induced by 20 mg/kg of azoxymethane (AOM). Here we studied whether fasting-refeeding could also affect 1) the induction of colon cancer by the same dose of AOM and 2) the induction of foci by lower doses of AOM that do not induce foci in fully fed rats. Fully fed and fasted-refed rats were given AOM by single subcutaneous injection, and the development of foci or tumors was evaluated three months or one year later. The results of the long-term carcinogenesis experiments showed that the total incidence of tumors was increased in the fasted-refed rats. Moreover, although fully fed rats developed foci only when injected with 7.5, 10, or 20 mg/kg of the carcinogen, a significant number of foci were also induced by 5 mg/kg in fasted refed rats. The crypt multiplicity of foci was also higher when rats were exposed to fasting-refeeding, even when the number of foci was unchanged. These data suggest that growth perturbations induced by fasting-refeeding lead to the development of preneoplastic lesions with doses of AOM too low to trigger foci in fully fed rats and produce enhanced sensitivity to the development of intestinal tumors. PMID- 10693168 TI - High dietary iron enhances oxidative stress in liver but does not increase aberrant crypt foci development in rats with low vitamin E status. AB - The purpose of this study was to examine the effects of high-iron and low-vitamin E diets on lipid peroxidation and aberrant crypt foci (ACF) development in rats. In a 2 x 2 x 2 factorial design, male Sprague-Dawley rats were fed 45 or 450 mg Fe/kg diet (adequate and high iron, respectively) and 15 or 100 IU vitamin E/kg diet (low and adequate vitamin E, respectively) for three weeks, when they received saline or azoxymethane (15 mg/kg for 2 wk). Diets were continued for an additional six weeks. Serum alpha-tocopherol concentrations in rats fed low vitamin E diets were decreased to 30% of concentrations observed in rats fed adequate-vitamin E diets (p < 0.0001). Also, serum alpha-tocopherol concentrations tended to be lower in rats supplemented with iron (p < 0.08). Lipid peroxidation in liver was significantly elevated by high-iron diets after 3 and 10 weeks of treatment, but lipid peroxidation in colonic mucosa was not altered by dietary iron or vitamin E. The total number of ACF and number of large ACF (> or = 4 aberrant crypts/focus) were not significantly altered by iron or vitamin E intakes. However, the size distribution of ACF was slightly altered, such that iron-supplemented rats had 12% more ACF with two crypts per focus (p < 0.02) than rats fed adequate-iron diets. Our data suggest that high-iron diets enhanced oxidative stress in liver, but not colon, of rats fed low-vitamin E diets. Furthermore, a high-iron diet does not increase the total number of ACF, even when vitamin E status is low. PMID- 10693169 TI - A probiotic strain of L. acidophilus reduces DMH-induced large intestinal tumors in male Sprague-Dawley rats. AB - Probiotic bacteria strains were examined for their influence on 1,2 dimethylhydrazine (DMH)-induced intestinal tumors in 100 male Sprague-Dawley rats. Lactobacillus acidophilus (Delvo Pro LA-1), Lactobacillus rhamnosus (GG), Bifidobacterium animalis (CSCC1941), and Streptococcus thermophilus (DD145) strains were examined for their influence when added as freeze-dried bacteria to an experimental diet based on a high-fat semipurified (AIN-93) rodent diet. Four bacterial treatments were compared: L. acidophilus, L. acidophilus + B. animalis, L. rhamnosus, and S. thermophilus, the bacteria being added daily at 1% freeze dried weight (10(10) colony-forming units/g) to the diet. Trends were observed in the incidence of rats with large intestinal tumors for three treatments: 25% lower than control for L. acidophilus, 20% lower for L. acidophilus + B. animalis and L. rhamnosus treatments, and 10% lower for S. thermophilus. Large intestinal tumor burden was significantly lower for treated rats with L. acidophilus than for the control group (10 and 3 tumors/treatment group, respectively, p = 0.05). Large intestinal tumor mass index was also lower for the L. acidophilus treatment than for control (1.70 and 0.10, respectively, p < 0.05). Other treatments showed no statistically significant change from control for these indexes of tumorigenesis. For rats fed L. acidophilus, no adenocarcinomas were present in the colons. Pulsed-field gel electrophoresis of bacterial chromosomal DNA fragments was used to differentiate introduced (exogenous) bacterial strains from indigenous bacteria of the same genera present in the feces. Survival during gut passage and displacement of indigenous lactobacilli occurred with introduced L. acidophilus and L. rhamnosus GG during the probiotic treatment period. However, introduced strains of B. animalis and S. thermophilus were not able to be isolated from feces. It is concluded that this strain of L. acidophilus supplied as freeze-dried bacteria in the diet was protective, as seen by a small but significant inhibition of tumors within the rat colon. PMID- 10693170 TI - Effects of garlic oil and its organosulfur compounds on the activities of hepatic drug-metabolizing and antioxidant enzymes in rats fed high- and low-fat diets. AB - We examined the effects of garlic oil (GO) and two of its organosulfur compounds, diallyl sulfide (DAS) and diallyl disulfide (DADS), on the drug-metabolizing and antioxidant systems in rats and sought to determine whether these effects are associated with dietary fat. Rats were fed a high-fat diet and received GO or DADS (200 mg/kg body wt) or DAS (100 mg/kg) orally three times a week for seven weeks. Control animals received corn oil alone. Another group of rats was fed a low-fat diet, with or without GO. GO and DADS significantly reduced the body weight gain of rats (p < 0.05). GO, however, dramatically increased the spleen weight and spleen weight-to-body weight ratio (p < 0.05). DAS increased glutathione S-transferase (GST) and 7-pentoxyresorufin O-dealkylase activities, whereas DADS increased only GST activity (p < 0.05). Immunoblot assay showed GO-, DAS-, and DADS-enhanced expression of the placental form of GST and cytochrome P 450 IIBI but suppressed cytochrome P-450 IIEI expression. Hepatic antioxidant enzyme activities were also modulated by these garlic components. GO and DADS inhibited glutathione peroxidase activity (p < 0.05), and DADS and DAS enhanced glutathione reductase activity (p < 0.05). Only GO enhanced the superoxide dismutase activity (p < 0.05). All these garlic components increased glutathione levels in red blood cells (p < 0.05) but did not influence hepatic glutathione levels. Although the amount of fat in the diet modulated drug-metabolizing and antioxidant functions, no interactions between GO and dietary fat were observed. These results indicate that GO and its allyl sulfide components, as well as dietary lipid, modulate drug-metabolizing and antioxidant enzyme activities. The action of GO appears to be independent of dietary lipid content. PMID- 10693171 TI - Genistein inhibits NF-kappa B activation in prostate cancer cells. AB - Prostate cancer is the second leading cause of cancer-related deaths in men in the United States. Epidemiological studies indicate that susceptibility to prostate cancer may be partly due to environmental influences, especially diet. An association has been shown between decreased prostate cancer risk and mortality with increased consumption of soy products, resulting in increased levels of isoflavones. We previously demonstrated that the soy isoflavone genistein inhibits cell growth and induces apoptosis in prostate cancer cells. To further elucidate the molecular mechanism by which genistein elicits its apoptotic effect, we investigated the role of a transcription factor, nuclear factor-kappa B (NF-kappa B), in the androgen-sensitive cell line LNCaP and the androgen-insensitive cell line PC3. Here we show that genistein decreases NF kappa B DNA binding and abrogates NF-kappa B activation by DNA-damaging agents, H2O2 and tumor necrosis factor-alpha, in prostate cancer cells regardless of androgen sensitivity. Additionally, we have demonstrated that genistein reduces phosphorylation of the inhibitory protein I kappa B alpha and blocks the nuclear translocation of NF-kappa B, prohibiting DNA binding and preventing NF-kappa B activation. These results provide a mechanism by which genistein induces apoptosis in prostate cancer cells: the inactivation of NF-kappa B. Furthermore, genistein's ability to abrogate NF-kappa B activation by DNA-damaging agents strongly supports genistein's role as a chemopreventive agent. PMID- 10693172 TI - Quercetin inhibits benzo[a]pyrene-induced DNA adducts in human Hep G2 cells by altering cytochrome P-450 1A1 gene expression. AB - Quercetin is one of the most abundant of the naturally occurring flavonoids. It has been estimated that about 25-50 mg of quercetin are consumed from the daily diet. The chemopreventive effect of quercetin on dietary carcinogen has been intensely studied in animal models; however, knowledge regarding the molecular mechanism is still limited. In this study, the human hepatoma Hep G2 cell line was used to investigate how quercetin prevents benzo[a]pyrene (B[a]P)-induced DNA adducts. The Hep G2 cells were treated with 10 microM B[a]P for 18 hours in the presence or absence of quercetin. The DNA adduct levels, evaluated by 32P postlabeling, decreased in a dose-dependent manner after treatment with quercetin. Cytochrome P-450 1A1 (CYP1A1) and glutathione S-transferase involvement have been well demonstrated in the modulation of B[a]P-induced DNA damage. From the assays of both enzyme activities, quercetin inhibits CYP1A1 linked ethoxyresorufin O-dealkylase activity more effectively than glutathione S transferase activity. To elucidate the molecular mechanisms, reverse transcriptase-polymerase chain reaction and Western blot were used to evaluate whether the decrease in CYP1A1 enzyme activity by quercetin is mediated because of alterations of CYP1A1 transcription or mRNA stability. The results indicated that quercetin significantly inhibits B[a]P-induced CYP1A1 mRNA and protein expression. From these findings, we conclude that quercetin suppresses B[a]P induced DNA damage in human Hep G2 cells by altering CYP1A1 gene expression. Thus we suggest that dietary quercetin may have a long-term preventive effect on chemical carcinogenesis, especially in people who eat a diet rich in fruits and vegetables. PMID- 10693173 TI - Role of nitric oxide and peroxynitrite in bile salt-induced apoptosis: relevance to colon carcinogenesis. AB - Previous work from our laboratory indicated that the bile salt sodium deoxycholate (NaDOC) induced apoptosis in cultured cells and in normal goblet cells of the colonic mucosa. We also reported that the normal-appearing flat mucosa of patients with colon cancer exhibited apoptosis resistance. Using immunofluorescence in conjunction with confocal microscopy, we now report that high physiological concentrations (0.5 mM) of NaDOC result in the formation of nitrotyrosine residues, a footprint for the formation of reactive nitrogen species, including peroxynitrite, in plasma membrane-associated proteins of HT-29 cells. Because peroxynitrite is formed from the reaction between nitric oxide and superoxide anion, we specifically looked at the role of nitric oxide and superoxide anion in NaDOC-induced apoptosis. Pretreatment of cells with the inhibitor/antioxidants, N-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase, copper (II) 3,5-diisopropyl salicylate hydrate, a superoxide dismutase mimetic compound, and Trolox, a water-soluble analog of alpha tocopherol, alone or in combination, sensitized cells to apoptosis induced by 0.5 mM NaDOC. These results suggest that nitric oxide may be part of a signaling pathway that is responsible for apoptosis resistance. The results also indicate that nitric oxide does not appear to protect cells against NaDOC-induced apoptosis by scavenging superoxide anion. PMID- 10693174 TI - Vitamin E (d-alpha-tocopheryl succinate) decreases mitotic accumulation in gamma irradiated human tumor, but not in normal, cells. AB - Previous studies have shown that treatment of tumor cells in vitro with d-alpha tocopheryl succinate (alpha-TS), a most effective form of vitamin E, alone or in combination with X-irradiation, reduced the growth of these cells more than that produced by individual agents. However, it is unknown whether alpha-TS, alone or in combination with gamma-irradiation, would produce similar effects on normal cells. To study this, we have compared the effects of alpha-TS on three human tumor cell lines, HeLa (cervical carcinoma), OVGI (ovarian carcinoma), and A549 (lung carcinoma), with the effects on three human normal fibroblast lines, GM2149, AG1522, and HF19. Results showed that alpha-TS treatment of HeLa cells for 20 hours caused inhibition of growth in a dose-dependent manner, but normal human fibroblasts treated similarly with alpha-TS did not show such an effect. alpha-TS treatment for 20 hours also decreased mitotic accumulation in all three tumor cell lines but did not produce such an effect in any of the normal fibroblasts. As expected, gamma-irradiation with 1 Gy decreased mitotic accumulation in human tumor cells and normal fibroblasts; however, alpha-TS treatment for 24 hours before, during, and after irradiation for the entire experimental period further decreased mitotic accumulation in human tumor cells but not in normal cells. These data suggest that effects of alpha-TS, alone or in combination with gamma-irradiation, are selective for tumor cells. Therefore, existing fear that antioxidants such as vitamin E may protect cancer cells from free radical damage during radiation therapy is not justified. PMID- 10693175 TI - Methylcobalamin decreases mRNA levels of androgen-induced growth factor in androgen-dependent Shionogi carcinoma 115 cells. AB - Methylcobalamin (MeCbl) is an important enzyme cofactor required for methionine synthase activity. It also inhibits, in a dose-dependent manner, the proliferation of an androgen-dependent cell line, SC-3, derived from an androgen dependent mouse mammary tumor (Shionogi carcinoma 115). In SC-3 cells, androgen induces the production of androgen-induced growth factor (AIGF), an autocrine growth factor increasing the proliferation of SC-3 cells. MeCbl treatment suppressed the androgen-induced, AIGF-mediated growth of SC-3 cells, as well as the androgen-induced increase of AIGF mRNA. In SC-3 cells, androgen receptors linked with androgen form complexes that tightly bind DNA and act as transcription factors in the nucleus to regulate the expression of specific genes such as AIGF. The number and dissociation constants of androgen receptors in control and MeCbl-treated SC-3 cells were the same. Similarly, the extent of binding of normal androgen receptors in nuclei from control and MeCbl-treated cells was virtually identical. The androgen receptors from control and MeCbl treated cells showed similar capacities for conversion to a form that tightly binds to DNA on heat activation. These results suggest that the reduction of AIGF mRNA, subsequent to the nuclear binding of androgen receptors, may be a partial cause of the growth-inhibitory activity of MeCbl in SC-3 cells. PMID- 10693176 TI - Glucocorticoid blockade does not abrogate tumor-induced cachexia. AB - Cancer-induced cachexia is a common manifestation observed in patients with malignancies. Elevated levels of circulating glucocorticoids and interleukin-6 (IL-6) have been observed in cancer patients with cachexia and are implicated as major mediators in this process. The purpose of this study was to investigate the role of circulating glucocorticoid levels as primary mediators in cancer-induced cachexia. We evaluated whether inhibition of glucocorticoids with the receptor antagonist RU-486 could abrogate the detrimental wasting of muscle and adipose tissues seen in a well-characterized murine tumor-induced cachexia model. Mice (12/group) were randomized to control, tumor-bearing, control + vehicle, or tumor bearing + glucocorticoid receptor antagonist groups. Circulating serum glucocorticoid and IL-6 levels were measured in addition to multiple body composition parameters, such as total body weight, lean body mass, and adipose content. The results of this study indicate a significant physiological alteration in the tumor-bearing host that causes severe and detrimental changes in body composition parameters. Regression analysis demonstrated a significant correlation between increased circulating glucocorticoid levels and alterations in body composition parameters. These observed defects were not abrogated with the administration of a glucocorticoid receptor antagonist. We therefore conclude that the untoward effects of tumor-induced cachexia are not mediated primarily by the peripheral effects of high circulating glucocorticoid levels but may involve a complex interaction with IL-6. PMID- 10693177 TI - Plant polyphenols inhibit benzoyl peroxide-induced superoxide anion radical production and diacylglyceride formation in murine peritoneal macrophages. AB - Naturally occurring plant polyphenols, which include ellagic acid (EA), tannic acid (TA), caffeic acid (CA), and ferulic acid (FA), were tested for their superoxide anion radical (SOR)-scavenging activities. SOR were produced by interaction of the tumor promoter benzoyl peroxide (BPO) with murine peritoneal macrophages in vitro. The levels of SOR were assessed microscopically by counting the number of formazan-positive cells per 250 cells produced by the reduction of nitro blue tetrazolium. BPO at a concentration of 15 micrograms/1.85 x 10(6) cells/0.5 ml induced maximum formation of SOR in resident and thioglycollate elicited cells. All the tested polyphenols were able to inhibit the formation of SOR induced by the tumor promoter to a variable degree. Inhibition of BPO-induced SOR formation by polyphenols was in the following order: FA > TA > CA > EA. BPO stimulated the accumulation of diacylglycerol (DAG) in resident and elicited macrophages with concurrent release of choline equivalents from macrophages. Polyphenols inhibited DAG accumulation, which paralleled the inhibition of choline equivalent release. FA was observed to be the most effective and EA the least effective inhibitor of SOR formation, DAG accumulation, and release of choline equivalents. It is likely that inhibition of SOR formation might be due to some interference in the cellular lipid metabolism and phospholipid equivalent deacylation and choline release. PMID- 10693178 TI - High intake of specific carotenoids and flavonoids does not reduce the risk of bladder cancer. AB - An analysis of a previously completed Spanish multicentric case-control study of bladder cancer was carried out using new available data on the contents in foods of specific carotenoids (alpha-carotene, beta-carotene, lutein, and lycopene) and flavonoids (quercetin, kaempferol, myricetin, and luteolin) to investigate the relationship of these phytochemicals with bladder cancer. The study included 497 cases first diagnosed with bladder cancer, 547 neighborhood controls, and 566 hospitals controls, matched by gender, age, area of residence, and hospital. Usual food intake was estimated using a dietary history questionnaire administered by trained interviewers. None of the specific carotenoids and none of the specific flavonoids have been found to be significantly associated with bladder cancer risk in this analysis. The adjusted odds ratios for subjects in the highest quartile of intake with respect to subjects in the lowest quartile were 1.36 (95% confidence interval = 0.94-1.95) for total carotenoid intake and 1.23 (95% confidence interval = 0.85-1.79) for total flavonoid intake. The results of this study does not support the hypothesis that intake of specific carotenoids and flavonoids is protective against bladder cancer risk. PMID- 10693179 TI - Examination of the adolescent patient. AB - Adolescent patients need knowledge and motivation to practice a healthy lifestyle. The provider of adolescent health care is uniquely qualified to provide factual health information and practical advice. Enlisting parental support for confidential adolescent health services usually is not problematic when parents and health care providers share common goals and responsibility. The health care provider must develop rapport to foster high-risk health behavior disclosure and must promote health messages that are stronger than those received from peers, television, movies, and magazines. Adolescents who elect to abstain from sexual activity need as much support as sexually active patients. Depression, substance abuse, and eating disorders must be recognized and treated. Preventative health care services for adolescents can be optimized when office staff understand the special needs of these young women. The physician's concerns regarding the health of adolescent patients will be welcomed by patients, their parents, and the community. PMID- 10693180 TI - Breast disorders in the pediatric and adolescent patient. AB - Despite the wide range of breast abnormalities that affect patients in the pediatric and adolescent populations, some conclusions can be drawn. Breast self examination in the adolescent population is controversial but is recommended for girls who carry the BRCA1 or BRCA2 gene beginning at age 18 to 21 years. All girls with a disorder of breast size or symmetry should be given the opportunity of consultation with a plastic surgeon to discuss reconstructive options. Ultrasound is the most appropriate initial investigation in any adolescent patient with a breast mass owing to the dense nature of breast tissue in this age group. Although it is extremely rare in this population, breast cancer must always be included in the differential diagnosis of a breast mass, particularly in the patient with a prior history of childhood malignancy or chest irradiation. PMID- 10693181 TI - Vulvar disorders in adolescents. AB - Vulvar disorders are rare in the adolescent. When they occur, they may be challenging and difficult to diagnose and treat. Biopsies and consultation with a dermatologist in unclear cases should occur early in the evaluation to allow for rapid diagnosis and appropriate treatment. PMID- 10693182 TI - Congenital malformations of the genital tract. AB - Congenital abnormalities of the genital tract are a source of major morbidity for teenage girls. Careful counseling is as important as the surgical approach to management. It is imperative that specialist centers are established worldwide to deal with these problems. Centralization of services will ensure these girls receive the highest quality care. PMID- 10693183 TI - Abnormal uterine bleeding in adolescents. AB - Irregular bleeding is a common complaint of adolescents and is responsible for approximately 50% of gynecologic visits in that age group. Most abnormal bleeding in adolescents is caused by immaturity of the hypothalamic-pituitary-ovarian axis resulting in anovulation. Approximately 20% of adolescents have an underlying disorder requiring targeted diagnostic testing. A thorough history and physical examination and laboratory findings will often reveal the source of abnormal bleeding. Appropriate treatment is then directed to the underlying cause. PMID- 10693184 TI - Disorders of excessive hair growth in the adolescent. AB - Virilization is most often the reflection of a serious underlying condition. Diagnosis and management should be prompt, thorough, and comprehensive. Hirsutism is the manifestation of a variety of disorders. It may be associated with serious acute medical conditions, chronic disorders, or idiopathic. The diagnosis should be methodical and adjusted to the nature of the clinical presentation. Several therapeutic modalities are effective and produce satisfactory results for most patients. PMID- 10693185 TI - Eating disorders in adolescents and young adults. AB - Eating disorders are relatively common and frequently result in gynecologic abnormalities. The gynecologist must appreciate the various manifestations of these complex health problems as well as the biopsychosocial approach needed to help the adolescent or young adult woman recover from these chronic conditions. By recognizing both the medical and gynecologic aspects of eating disorders, the oversimplified viewpoint of considering these conditions as purely psychiatric disorders can be avoided. Open and consistent communication with patients, with a focus on health rather than dysfunction and mental illness, facilitates the acceptance of a comprehensive approach involving the gynecologist, dietitian, and mental health provider. PMID- 10693186 TI - Female adolescent sexuality. Promoting healthy sexual development. AB - Health care providers must recognize the specific challenges and rewards of providing services for adolescents. Quality care begins with the establishment of trust, respect, and confidentiality between the health care provider and the adolescent. Data suggest that the normal age for beginning puberty is decreasing, which has important clinical, educational, and social implications. The health care provider should be aware of the broad range of potential sexual behaviors involving adolescents, as well as the teen's acceptance of such behaviors, often dictated by age, gender, culture, and education. When providing gynecologic care to adolescent girls, the physician should not only provide contraception and screen for sexually transmitted diseases but should contribute to the development of the patient's sexual health. Especially when providing care for the younger teen, the health care provider must focus on involving a member of the family or another significant adult to provide needed support and guidance. Anticipatory guidance for parents should focus on assessing their parenting styles and promoting supervision. Although parents should strive to maintain open communication with their adolescents, they may not accurately estimate the sexual activity of and the sexual risk for their teenage children. Parents need to be encouraged to consider the implications of their own sexual behaviors. The provider should attempt to foster a comfortable environment in which youth may seek help and support for appropriate medical care while reserving the right to disclose their sexual identity when ready. Health care professionals cannot exclude heterosexual behavior on the basis that a young woman self-identifies as homosexual. Her reported sexual behaviors may not indicate her sexual orientation. Self-definition of sexual orientation is a dynamic process including factors such as fantasies, desires, and behaviors. Self-definition of sexual identity is affected by individual variations in sex, gender, sexual roles, and sexual orientation. Most adolescents want to discuss sexual-related issues with their health care providers and will welcome direct questions about sexual behaviors and possible risks when posed in a confidential and nonmoralistic manner. Discussion of the physical, emotional, familial, and social changes related to adolescence will encourage healthy sexual development. PMID- 10693187 TI - Update on adolescent contraception. AB - Recent advances in OCPs include less androgenic progestins and lower doses of ethinyl estradiol. All low-dose OCPs are safe in terms of venous thrombosis risk in appropriately chosen patients. DMPA is a safe and effective long-acting contraceptive agent; clinical attention should be directed to its most common side effect, irregular bleeding. DMPA does not seem to affect mood, and it is uncertain what impact it has on weight changes. More research needs to be conducted on its impact on adolescent bone metabolism. Norplant continues to be the only subdermal contraceptive implant marketed in the United States. It provides safe and effective contraception and has the best continuation rate of all types of hormonal contraception. Its most common side effect is irregular bleeding. Norplant may be especially well suited for adolescents who have recently been pregnant or who are not tolerating other types of contraception. Emergency postcoital contraception continues to be underused in the United States, with a lack of awareness among patients and clinicians. Mechanisms of action include a delay in ovulation and interference with implantation. Research and public health groups are striving to increase patient and provider awareness and use of emergency contraception. PMID- 10693188 TI - Adolescents and sexually transmissible diseases. AB - Sexually transmitted infections are alarmingly common among adolescents in the United States. Behavioral, biologic, and health care access factors place adolescent girls at high risk for many common infections. This population also experiences a disproportionate burden related to the sequelae of STDs. The costs are high for the individual adolescent and for society. Clinicians treating adolescent girls should address the general lack of knowledge about the risks and consequences of STDs. They should be prepared to offer confidential and comprehensive counseling, screening, and treatment according to established guidelines. Office policies that protect adolescent confidentiality are an important component in providing effective care. Adolescence is a period during which lifelong health behaviors are established. It provides a critical opportunity for promoting responsible behaviors and reducing risks through health promotion and prevention strategies. PMID- 10693189 TI - Pelvic pain. A SAFE approach. AB - The efficacy of a multidisciplinary approach for the treatment of chronic pain has been well documented. A recent randomized prospective trial by Peters and co workers confirmed that multidisciplinary management of chronic pelvic pain resulted in greater improvement in pain severity, increased employment, and improved functional health scores when compared with a traditional medical approach. Unfortunately, many gynecologists do not have the luxury of working in these types of settings. The SAFE approach allows the busy practitioner to evaluate the nonorganic and organic nature of pelvic pain simultaneously to manage these difficult patients better. Initially, it is important to set up regular weekly visits with a prespecified time limit. During these sessions, one should focus on compliance to treatment recommendations. Components of management that have led to pain reduction should be defined. Over time, these short visits can be spaced to bimonthly and monthly intervals; however, this strategy should not be rushed. If possible, the gynecologist should coordinate the care of difficult patients with a physical therapist and mental health provider who can work in tandem on pain reduction strategies and focused psychotherapy. The gynecologist should focus the patient's attention on improvements in pain reports and in functioning at school or work and reinforce the consistent use of medications, even if the recommended dose is low. Small improvements in health beget larger improvements in daily functioning. PMID- 10693190 TI - Reproductive health services for adolescents. Critical legal issues. AB - The contemporary legal and policy environment has increased the challenges associated with providing health care services to the adolescent population. The issue of reproductive health care services is particularly intense because of the controversial nature of services for contraception and abortion. As the debates continue, one must remember the background against which they are occurring. The current legal framework, developed over nearly 40 years, enables adolescents who are minors to give their own consent for care in numerous circumstances and provides them with a significant level of confidentiality protection, particularly for reproductive health services. Laws have been enacted to expand adolescents' financial access to health care, through targeted publicly funded service programs and expanded health insurance coverage. This background provides the foundation for addressing the current challenges and for protecting and expanding adolescents' access to care. PMID- 10693191 TI - 'A real call to action'. PMID- 10693192 TI - Questioning APR cartridge use. PMID- 10693193 TI - Questioning APR cartridge use. PMID- 10693194 TI - Ten ways to deal with turf wars. PMID- 10693196 TI - Ergonomics E-sources. PMID- 10693195 TI - ASHRAE 62-1999 ventilation for acceptable indoor air quality, addendum n. PMID- 10693197 TI - Fresh ideas for keeping crews safe. PMID- 10693198 TI - Where workers need more than a brake. PMID- 10693200 TI - Before the fire. PMID- 10693199 TI - Small spaces, big hearts. PMID- 10693201 TI - Unseen dangers. PMID- 10693202 TI - Looming liability. PMID- 10693203 TI - The S.P.I.C.E. model for return to work. PMID- 10693204 TI - The aggression continuum: a paradigm shift. PMID- 10693205 TI - Don't forget the "D" in the "ABCs" of emergency care. PMID- 10693206 TI - Emergency cases with a twist. PMID- 10693207 TI - A teenager acting strangely. PMID- 10693208 TI - Bronchiolitis--or is it? PMID- 10693209 TI - Hocus-pocus: a case of abdominal pain after blunt abdominal trauma. PMID- 10693210 TI - An offbeat wheezer. PMID- 10693211 TI - A lethargic infant: ingestion or deception? PMID- 10693212 TI - A 12-year-old boy with protean pain. PMID- 10693213 TI - The case of the red eye. PMID- 10693214 TI - The case of the missing "olive". PMID- 10693215 TI - Getting to the heart of the matter. PMID- 10693217 TI - Ageing and safety promotion--what do we know and where are we going? PMID- 10693216 TI - Health promotion for older people: what are the opportunities? PMID- 10693218 TI - Dementia care: challenges for an ageing Europe. PMID- 10693219 TI - Planning a breast cancer health promotion. Qualitative and quantitative data on Puerto Rican elderly women. PMID- 10693220 TI - The WHO perspective on active ageing. PMID- 10693221 TI - Brief to the World Trade Organization: world trade and population health. PMID- 10693222 TI - [Gerontology in the health promotion world]. PMID- 10693223 TI - Commentary: beyond preclassified reality. PMID- 10693224 TI - Fabricated trauma exposure: an analysis of cognitive, behavioral, and emotional factors. AB - The 1995 Oklahoma City bombing was a disaster of unparalleled dimension in the United States. The professional response included the development of systematic clinical and research programs. This article describes the case of a child who, as a participant in a research study, appeared to fabricate a story of bomb related loss. The research and clinical records of this child were examined and analyzed according to the factors and conditions that might underlie this fabrication. These include issues related to memory and suggestibility, symptom contagion, and mass hysteria. The report describes the role of psychological vulnerability in trauma and this child's coping and adaptation. PMID- 10693225 TI - Avoidance in trauma: conscious and unconscious defense, pathology, and health. AB - Drawing from our work with children seen following the 1995 bombing of the Alfred P. Murrah Federal Building in Oklahoma City, Oklahoma, this article describes clinical aspects of avoidance in traumatized children and their families. Avoidance in traumatized children and their families seems a final common pathway arising from a number of diverse factors. The importance of particular factors for assessment and treatment is emphasized. PMID- 10693226 TI - Posttraumatic obsessive-compulsive disorder: a three-factor model. AB - This paper presents a three-factor causal model of obsessive-compulsive disorder (OCD), which posits that exposure to long-term traumatic stress generates an inordinate degree of anxiety during the psychological development of the premorbid OCD child. In response to these conditions the child evolves a distinct cognitive style characterized by exaggerated threat appraisal and magical beliefs, and experiences alterations in brain metabolism. An entire functional brain system (a basal ganglia-orbitofrontal circuit) enters into a state of enhanced responsiveness following exposure to protracted threat. Over time the threshold for stimulation is dramatically lowered, resulting in a hypersensitivity to cues that signify potential harm. Individuals adapt to this hypersensitivity through a variety of strategies, which constitute OCD. PMID- 10693227 TI - Posttraumatic stress disorder: cerebellar regulation of psychological, interpersonal, and biological responses to trauma? AB - Alteration in the sense of time is the most commonly reported peritraumatic dissociative symptom. A case report of a trauma victim illustrates the posttraumatic alteration in the sense of time as well as loss of spatial memory. Recent studies of cerebellar function indicate the cerebellum may be critical to both spatial memory and the sense of time. Identifying regulators of psychological, interpersonal, and biological responses to traumatic events is important in advancing our understanding of the effects of trauma. The cerebellum may be part of the initial posttraumatic response. PMID- 10693228 TI - Commentary: deconstructing self-destruction. PMID- 10693229 TI - "You are not your body": commentary on "The motivations for self-injury in psychiatric inpatients". PMID- 10693230 TI - The motivations for self-injury in psychiatric inpatients. AB - Nonsuicidal self-injurious behavior (SIB) occurs in both culturally appropriate and culturally inappropriate forms. It is one of the diagnostic criteria for borderline personality disorder, but it occurs in several psychiatric and neurological populations. The personal intent of SIB in psychiatric populations is incompletely understood. A self-report scale (Self-Injury Motivation Scale; SIMS) to assess motivation for self-injury was developed. Relationships among motivation for SIB, characteristics of SIB, and psychopathology were explored. A semistructured interview and the SIMS, Dissociative Experiences Scale, Beck Depression Inventory, Davidson Trauma Scale, and Millon Clinical Multiaxial Inventory-II were given to 99 consecutively admitted inpatients. The SIMS had good reliability and validity. A high SIMS score suggested distinct psychopathology. Several factors on the SIMS differentiated motivations for SIB. Patients with different SIMS factor profiles had different psychopathology. PMID- 10693231 TI - Family boundary ambiguity predicts Alzheimer's outcomes. AB - This study examines caregiver and patient relationship characteristics in the etiology of behavior problems in Alzheimer's disease. Seventy-two caregivers and patients were assessed twice, 12 months apart. Cross lag panel analysis was used to test for one-way or reciprocal causal links among caregiver variables, patient impairment measures, and patient behavior problems. Caregiver distancing from patients (closeout) predicted increases in the frequency of behavior problems, including activity disturbances, paranoia, and anxiety. These behaviors in turn led to increased closeout of the patient by the caregiver. The reciprocal causal associations found in this study suggest that dysfunctional family interactions may underlie patient behavior problems and caregiver distress. PMID- 10693232 TI - Adjunctive cognitive-behavioral therapy for rapid-cycling bipolar disorder: an empirical case study. AB - A basic biopsychosocial model of episode onset in rapid-cycling bipolar disorder is presented with a special emphasis on cognitive and other psychosocial contributors. A three-pronged, face-valid, cognitive-behavioral treatment protocol meant to supplement medications is deduced from the available research literature. The concrete treatment components focus on prevention of mood cycles, early detection of cycle onset, and mood restabilization during cycles. The treatment protocol was pilot tested on a rapid-cycling bipolar patient who first received pharmacotherapy only followed by pharmacotherapy plus adjunctive cognitive-behavioral therapy (CBT). Detailed treatment measures were collected before, during, and after treatment. A comparison of Beck Depression Inventory and Young Mania Scale scores in pharmacotherapy versus pharmacotherapy plus CBT conditions suggest the addition of CBT produces significant clinical gains. Scores on the Beck Anxiety Inventory and Hopelessness Scale provide further support for the CBT treatment model. These preliminary results suggest cognitive behavioral or similarly structured psychosocial treatment models could greatly enhance the medical treatment of rapid-cycling bipolar patients and warrants further controlled investigation. PMID- 10693233 TI - Exploring the relationship between the person and the disorder among individuals hospitalized for psychosis. AB - Recent research has suggested that people suffering from severe mental illness develop strategies to cope with their disorder. This study describes one specific strategy of coping with psychosis--through the use of narrative or a story. The report is based on bimonthly, semistructured research interviews conducted with 43 persons who were hospitalized for severe mental illness with psychotic features over a 1-year period. Qualitative analysis revealed five distinct categories, which seem to reflect different interactions between the participants' sense of self in relation to their illness. Narrative theory helped to identify subtleties in these interactions described by participants and conceptualize the categories. Conceptual and clinical implications of this process and future research opportunities are discussed. PMID- 10693234 TI - Epidemiology of esophageal cancer, especially adenocarcinoma of the esophagus and esophagogastric junction. AB - The incidence of adenocarcinoma of the esophagus and esophagogastric junction (EGJ) has been increasing over the past 15 years in western countries. Surgical series and population-based studies show that, by 1994, adenocarcinomas of the esophagus accounted for half of all esophageal cancer among white men. The causes of this increase in incidence remain to be elucidated. Esophageal adenocarcinomas and a portion of EGJ adenocarcinomas arise from long and short segments of specialized intestinal metaplasia (Barrett's esophagus). The prevalence of long segments of Barrett's esophagus (> 3 cm) in patients having endoscopy for reflux symptoms is 3%, and 1% in those undergoing endoscopy for any clinical indication. However, a silent majority of patients with Barrett's esophagus remain unrecognized in the general population and may not be diagnosed unless adenocarcinoma develops. Recent studies document a rise in the diagnosis of specialized intestinal metaplasia of the cardia. Nearly all these patients have associated carditis, and Helicobacter pylori infection has been linked to this condition. The possible origin of EGJ adenocarcinomas in the sequence carditis- specialized intestinal metaplasia needs to be clarified. Smoking and obesity are additional risk factors for adenocarcinoma of the esophagus and EGJ. Current data does not confirm H. pylori as a risk factor for cancer of the EGJ. PMID- 10693235 TI - Modern pathology: prognostic parameters in squamous cell carcinoma of the esophagus. AB - In the present study the prognostic impact of new histological and molecular parameters were tested retrospectively in a series of 149 patients with esophageal squamous cell carcinoma (SCC) who underwent potentially curative resection therapy. In addition, the prognostic value of various molecular markers was investigated in a group of 38 patients with locally advanced esophageal SCC treated using combined therapy modalities. In the surgically treated carcinomas, the following morphological parameters proved to be prognostically significant in univariate survival analysis and multivariate survival analysis: pattern of invasion, inflammatory response, and lymph vessel invasion. In contrast, tumor grading according to the criteria of the WHO and tumor cell proliferation did not show significant prognostic impact. Concerning the prognostic influence of molecular parameters, strong expression of the proliferation regulating molecule p21WAF1 and weak expression of the apoptosis regulating molecule Bcl-XL were predictors of poor survival in univariate and multivariate survival analysis. No prognostic impact was shown in relation to the expression of p53 and the apoptosis regulating molecules Bcl-2 and Bax. In the multimodally treated esophageal cancer patients, strong expression of p21WAF1 and accumulation of p53 were predictors of poor survival, whereas expression of Bcl-2, Bax, and Bcl-XL had no prognostic significance. In conclusion, morphological and molecular parameters may provide important prognostic information for esophageal cancer patients. PMID- 10693236 TI - Malignant progression in Barrett's esophagus: pathology and molecular biology. AB - Due to its increasing incidence, esophageal adenocarcinoma and its precursor lesions have received increasing attention in recent years. The histopathologic steps in the process of malignant progression in Barrett's esophagus are well described and include the following: (a) metaplasia of the normal esophageal squamous epithelium to a specialized intestinal glandular epithelium, (b) development of dysplasia (classified histologically as low and high grade), and (c) development of adenocarcinoma characterized by invasive and metastatic potential. Intestinal metaplasia can be identified by the presence of goblet cells, the detection of which can be aided by finding mucin stained by Alcian blue at low pH. Despite this well-characterized sequence, the timing of the development of dysplasia and the subsequent transition to carcinoma and the risk of development of carcinoma in low- and high-grade dysplasia are not precisely known. In addition, there are problems in the identification of dysplasia, including sampling error and interobserver discrepancies among pathologists. A better understanding of the mechanisms of these events would allow early identification and elimination of high-risk lesions before adenocarcinoma with its attendant poor prognosis were able to develop. In order to better understand this process and to potentially identify early markers of malignant transformation, a variety of molecular studies have been carried out in recent years on adenocarcinoma and its precursor lesions in Barrett's esophagus. On the phenotypic level, increased expression and changes in pattern of expression of proliferation marker (Mib-1) Ki-67 antigen, overexpression of p53 protein, overexpression of growth factors such as epidermal growth factor (EGF), c-erbB2, and transforming growth factor (TGF)-a, decreased and abnormal expression of the cell adhesion molecule E-cadherin, and, in carcinomas, increased expression of serine proteases have all been described. A new area of interest is the family of rab proteins, which play an important role in maintaining cell polarity in the gastrointestinal tract. Increased expression of one of these proteins, rab11, has been described in low-grade, but not high-grade dysplasia. In cytogenetic studies, an increased S-phase fraction, followed by an increased tetraploid (4N) fraction and then aneuploidy, has been described. So far, the specific genes which have been most thoroughly investigated have been p53, APC, p16, and the sites of probable tumor suppressor genes, including 3p (FHIT), 13q, and 18q. With only a few exceptions (i.e., rab11 expression, and possibly mutations of FHIT), the numerous molecular abnormalities which have been described occur late in malignant progression, which means that the best marker which presently exists to identify high-risk lesions in Barrett's esophagus is the histologic identification of dysplasia in endoscopic biopsies, especially high-grade dysplasia. We are presently beginning studies using laser microdissection and competitive genomic hybridization (CGH), which could help to identify new chromosomal areas that might contain genes that are crucial in the early phases of malignant progression in Barrett's esophagus. In the future, identification of such early molecular events which predispose to carcinoma development will allow more precise and earlier risk assessment for individual patients, therefore, enabling more effective therapy. PMID- 10693237 TI - Malignant degeneration of Barrett's esophagus: clinical point of view. AB - The incidence of adenocarcinoma of the distal esophagus is increasing at an alarming rate. Intestinal metaplasia in the distal esophagus, i.e. Barrett's esophagus, has been identified as the single most important risk factor for these tumors. Barrett's esophagus develops as a consequence of chronic mucosal injury in up to 10% of patients with long-lasting gastroesophageal reflux disease. Experimental and clinical data indicate that adenocarcinoma of the distal esophagus is a direct consequence of mixed (i.e., acid and bile) reflux into the esophagus. Interestingly, Helicobacter pylori infection of the stomach appears to exert a protective effect against the development of esophageal adenocarcinoma. Neither aggressive medical acid suppression nor antireflux surgery can induce a predictable regression of Barrett's esophagus or exert a protective effect against its malignant degeneration. Endoscopic ablation of Barrett's esophagus, although appealing, currently constitutes a potentially dangerous procedure without proven benefit for the patient. Since the development of Barrett's adenocarcinoma follows a multistep process from metaplasia through increasingly severe grades of dysplasia, close endoscopic surveillance with extensive biopsies currently remains the only means to identify patients at risk for malignant degeneration and detect esophageal adenocarcinoma at an early and curable stage. PMID- 10693238 TI - New diagnostic methods for esophageal carcinoma. AB - The increasingly severe problem of esophageal carcinoma on world public health merits the application of new endoscopic methods to assist in early detection and screening. Older methods, such as tissue staining, combined with magnification endoscopy, have shown promising results, while newer techniques capitalize on measurements that discriminate benign from malignant cells based on a wide array of different attributes, ranging from the molecular to the macroscopic level. Instrumentation based on laser-induced fluorescence spectroscopy, ratio fluorescence imaging, elastic scattering spectroscopy, Raman spectroscopy, and optical coherence tomography is presently being tested and compared with standard endoscopic techniques. Using pathologic interpretation of pinch biopsies as the "gold standard," these techniques have shown the ability to identify dysplastic or malignant regions of tissue that would not be visible to the unassisted endoscopist and offer increased sensitivity for detection compared to rigorous random biopsy protocols. The rapid speed of the instruments allows the provision of information to the endoscopist almost instantaneously, potentially allowing therapeutic decisions to be conducted within the confines of the same endoscopic procedure, thereby achieving gains in efficiency and reductions in overall cost. Large, multicenter trials will be necessary to determine the sensitivity and specificity of individual and combined techniques, as well as their ability to favorably influence the early detection, management, and overall outcome of this disease. PMID- 10693239 TI - Esophageal carcinoma: current staging strategies. AB - A patient with suspected esophageal carcinoma represents a challenge to the treating physicians. Most patients present with an advanced stage of disease, and in the majority of cases only palliative treatment can be offered. Various treatment modalities are available, which are applied according to the TNM stage of the disease and the performance status of the patient. A precise histological diagnosis and highly accurate tumor staging of a patient with esophageal carcinoma is a prerequisite for the selection of the most suitable treatment option. Endoscopic ultrasound (EUS) has emerged as the most accurate diagnostic modality for locoregional staging. Problems in identifying early tumor stages or tumor strictures can be generally overcome by using miniprobe sonography (MPS). EUS/fine-needle aspiration biopsy (FNA) technology provides a valuable means of identifying suspicious locoregional lymph nodes. Patients with a proximal tumor (trachea bifurcation) should undergo bronchoscopy to rule out infiltration of the tracheobronchial system. Ultrasound (US), computed tomography (CT), and possibly magnetic resonance imaging (MRI) are the diagnostic tools of choice for extended tumor staging. After excluding extended tumor stage and severe concomitant diseases, diagnostic laparoscopy with intra-abdominal ultrasound should be performed in patients with adenocarcinoma of the esophagus prior to esophagectomy. Intra-abdominal metastases which can be missed preoperatively in some cases have to be ruled out in order to avoid unnecessary surgery. PMID- 10693240 TI - What's new in imaging? New magnetic resonance imaging of esophageal cancer using an endoluminal surface coil and antibody-coated magnetite particles. AB - An endoluminal surface coil: Esophageal cancer was studied by high resolution magnetic resonance imaging (MRI) to determine the histopathologic basis for signal intensity in these lesions and to determine the potential of this modality for evaluating the depth of cancer invasion. In a basic study, 14 tumors were examined with a 1.5-T superconductive MR system using a surface coil. The esophageal wall could be differentiated into four layers on the T1-weighted images and seven layers on the T2-weighted images. The signal intensity of the tumor varied from low to intermediate on the T1- and T2-weighted images. The submucosal layer is important in evaluating cancer invasion on the T2-weighted images. In a clinical study, 30 patients with esophageal cancer were examined with a 1.5-T superconductive MR system using an endoluminal surface coil. In terms of depth of cancer invasion, the accuracy rate of MRI using the endoluminal surface coil was 83%. In conclusion, MRI with an endoluminal surface coil will be a useful examination for esophageal cancer in the future. Antibody-coated magnetite particles: A highly specific and effective MRI contrast agent was prepared by coating superparamagnetite particles with monoclonal antibodies (MAbs) directed against epidermal growth factor receptors (EGFRs), which are overexpressed in esophageal squamous cell carcinoma. The agent was shown to have EGFR-specific MRI contrast capacity in vivo in athymic rats bearing TE8 or H69 tumors. Immunospecific MRI using magnetite particles coated with MAbs against EGFR seems to be useful in the diagnosis of squamous cell carcinoma of the esophagus. PMID- 10693241 TI - Risk analysis in esophageal surgery. AB - The postoperative mortality after esophagectomy still remains a major factor influencing the prognosis of esophageal cancer and largely depends on the patient's preoperative physiological status. A composite scoring system was developed to predict the risk of esophagectomy, based on quantitative assessment of preoperatively available physiological parameters. The scoring system was reviewed retrospectively on operated patients and evaluated prospectively in two subsequent patient groups. An initial retrospective multivariate analysis of 432 esophagectomy patients identified a compromised general status (p = 0.001) and poor cardiac (p < 0.001), hepatic (p < 0.05), and respiratory (p < 0.05) functions as independent predictors of a fatal postoperative course. Based on the relative risks associated with individual impaired organ functions--general status 3.6, cardiac function 2.8, hepatic function 2.1, pulmonary function 1.7--a composite risk score was established. A prospective study in 121 patients confirmed that this composite scoring system provides better identification of high-risk patients than does any of the individual parameters alone. Including this composite score into the process of patient selection and choice of procedure resulted in a decrease of postoperative mortality from 9.4% (52/553) to 1.2% (4/323) (p = 0.001). The risk of death after esophagectomy for esophageal cancer can be objectively assessed prior to surgery and quantified by a composite risk score. This score provides a useful tool in refining the criteria of patient selection for resection and choice of procedure, and markedly reduces postoperative mortality when applied prospectively. PMID- 10693242 TI - Neoadjuvant chemoradiation followed by surgery for resectable esophageal cancer. AB - Neoadjuvant chemoradiation (NAC) therapy protocols were developed to improve survival in patients with resectable esophageal cancer. Our experience with two consecutive NAC therapy trials is reviewed. Both studies included patients with localized squamous cell cancer and adenocarcinoma. Patients were treated with cisplatinum 26 mg/m2/day (days 1-5 and 26-30), 5-Fluorouracil (5-FU) 300 mg/m2/day (days 1-30), concurrent radiotherapy (4400 cGy) followed by esophagectomy. In the second trial, adjuvant taxol was added. The first protocol had 50 patients. Two patients died, both before surgery, one from sepsis. There was no residual viable tumor (CR) in 19 (40%) patients. The median survival time was 31 months. The 5-year survival rate of 36% compared favorably with concurrent 5-year survival of 18% for surgery alone. Forty-one patients were enrolled in the second trial. All underwent surgery. There were no treatment or operative deaths. Survival data for this group is maturing. Combined results from both protocols are: treatment mortality of 2.2%, complete response rate of 37%, and a median and 3-year disease-specific survival of 42 months and 54%, respectively. We conclude that NAC followed by surgery improves survival over surgery alone and that CR is predictive of improved survival. PMID- 10693243 TI - Neoadjuvant therapy of squamous cell carcinoma of the esophagus. AB - Because of the high risk of recurrence following surgery for squamous cell carcinomas of the esophagus, treatment involving both systemic therapy and a localized approach [either surgery alone or chemo/radiation and surgery] have been extensively studied. Pilot trials have demonstrated safety for the use of chemotherapy followed by operation, or for chemotherapy plus standard radiation plus surgery. Larger scale phase III trials, in which either of these two strategies is employed, have now been reported. For chemotherapy followed by surgery, conflicting results have recently been noted. A US intergroup trial showed no significant outcome for all patients treated. A smaller European study, also using a cisplatin-based regimen, had a positive result. Similarly, for chemoradiation followed by operation, two recent random assignment studies failed to demonstrate a significant difference in outcome. However, recently newer chemotherapy agents have been identified which hold promise for more effective therapy. These include paclitaxel and irinotecan. Phase II trials, using these agents prior to surgery, are underway or have been completed, and phase III studies are in the advanced planning stage. Until these trials have demonstrated superiority, surgery alone or chemoradiation without surgery remain the standard of care, for patients with esophageal cancer. PMID- 10693244 TI - Therapy of cervical esophageal carcinoma. AB - Surgical treatment of cervical esophageal cancer is influenced by special problems arising from the anatomical characteristics of this organ. The cervical and thoracic extension of these tumors makes an extensive lymphadenectomy necessary, and radical resections often may only be achieved by laryngectomy. The extent of the resections performed determines the type of intestinal restoration by gastric or colonic interposition and small bowel transplantation. The patient's voice may be preserved by means of tracheopharyngeal shunts with intestinal interposition. The advances of radiation therapy and chemotherapy will enable less extended resections with greater rates of laryngeal preservation. PMID- 10693245 TI - Esophageal cancer: a European perspective. AB - A survey was conducted among a group of European surgeons in order to investigate current attitudes and strategies in the management of squamous cell carcinoma of the esophagus. The survey consisted of a questionnaire mailed to surgeons in eight different countries with extensive clinical experience and scientific interest in the field. Eight questionnaires including the data of 6146 operated patients were available for analysis. A consensus emerged among the panelists that protocols of induction therapy should be routinely used in patients with locally advanced disease, especially in supracarinal tumors. Four of the surgeons advocated bilateral neck dissection in these patients. A progressive improvement in survival over the past three decades was noted. After 1990, the postoperative mortality rate was 6.2% after surgery alone and 9.7% after chemoradiation therapy followed by surgery. The 5-year survival rate after a complete resection was 38.5%. In the opinion of the panelists, esophagectomy remains the "gold standard" of therapy and should be regarded as an integral component of the treatment plan for patients with squamous cell carcinoma. PMID- 10693246 TI - Significance of extended systemic lymph node dissection for thoracic esophageal carcinoma in Japan. AB - In 1986, several institutions in Japan began to employ extensive lymphadenectomy for thoracic esophageal cancer. The aim of this article is to point out several confusing factors concerning the use of the terms "tow-field" and "three-field" lymph node dissection for thoracic esophageal cancer. In two-field nodal dissection, two components are included with (modern two-field) or without (traditional two-field) nodal dissection around both recurrent laryngeal nerve chains in the upper mediastinum. We studied a series of 353 patients resected for thoracic esophageal cancer in our institution. The patients were divided into three groups. Group A was the traditional two-field group of patients who underwent thoracoabdominal lymphadenectomy without upper mediastinal lymph node dissection after preoperative irradiation; group B was the modern two-field group, with additional upper mediastinal lymph node dissection; and group C was the three-field group with additional neck lymph node dissection. Groups B and C were operated on during the same period and did not received preoperative irradiation. The 5-year survival rate in group B was 54.9%, which was better than the 47.6% rate after three-field dissection (group C). The key to extensive lymphadenectomy for thoracic esophageal cancer does not lie in "cervical dissection" but in the meticulous dissection of the lymph nodes around the right and left recurrent laryngeal nerves. PMID- 10693247 TI - Gastric conditioning. AB - The etiology of esophagogastric anastomotic leaks is often multifactorial. However, occult ischemia of the gastric fundus is an important cause. In gastric conditioning, preliminary partial gastric devascularization is carried out 2-3 weeks before construction of the esophagogastric anastomoses. Gastric vascularity improves over this time. In animal studies, gastric conditioning has reduced the incidence of anastomotic leaks. Clinically, the concept of gastric conditioning can be used in several ways. Esophagectomy can be done at one stage, then a cervical esophagogastric anastomosis can be completed as a second-stage procedure. Pre-esophagectomy angiographic gastric artery embolization is another method of gastric conditioning. Finally, laparoscopic partial gastric devascularization can be done at the time of laparoscopic cancer staging. For gastric conditioning to be clinically useful, the benefit from reduction in leaks must be greater than the costs and morbidity of the conditioning procedure itself. PMID- 10693248 TI - Fundus rotation gastroplasty: a modified gastric tube. AB - Anastomotic failure remains to be one of the main problems in esophageal surgery with leakage rates up to 30% being reported currently. We addressed that problem by modifying the gastric-tube formation in utilizing all of the gastric fundus and omitting the resection of the lesser gastric curvature and accompanying vessels. Experimentally, those fundus-rotation gastroplasties were significantly longer and better perfused than conventional gastroplasties. In a clinical series of 53 patients (45 male, eight female, range 42-83 years) undergoing esophageal resection (49 malignancies, four non-malignant esophageal disease), four anastomotic leakages were found (7.5%). Three patients died due to a bronchial leak and adult respiratory distress syndrome, one with a complete gastric-tube necrosis and one with a colon perforation, sepsis and multiorgan failure. Eight patients developed a significant anastomotic stricture requiring repeated endoscopic dilatations. From our experimental and clinical experience, we conclude that the favorable length and perfusion of fundus-rotation gastroplasties allow for safe anastomosis at either the cervical or thoracic level. PMID- 10693250 TI - Complications following esophageal surgery. AB - Chronologically, complications can be classified as intraoperative, early, and late. The authors analyze complications according to this classification on the basis of more than 400 esophageal operations and related literary data. As regards intraoperative complications, they deal only with those occurring at transhiatal esophagectomy (e.g., tracheal tear, bleeding, pneumothorax, laryngeal nerve injury). Among the early complications, they survey the incidence of transplant necrosis and related mortality, further sequelae ensuing from subacute ischemia of the replaced organ and analyze in detail the questions which arise regarding anastomotic leakage. Firstly, they deal with those causative factors that influence the frequency of anastomotic insufficiency, such as the technical "know-how" of anastomosis making (e.g., one layer vs two layers; stapling or manual suture; interrupted or running suture), the way of replacement using whole stomach or tube-stomach and the consequences originating from the route of replacement (e.g., anterior or posterior mediastinal route). Incidence and management of chylothorax are also dealt with. While dealing with late complications, the authors give a detailed comment on anastomotic strictures and also other factors facilitating the development of late dysphagia, such as peptic stricture and tumor of the organ remnant. Finally, some cases successfully treated by surgery are presented (skin-tube formation in cases following transplant necrosis; abolition of a pharyngogastric anastomosis stricture using a free jejunal transplant and surgical solution of an anastomotic stricture from median sternotomy approach). PMID- 10693249 TI - Colon interposition. AB - In the anatomy of the colon vasculature, the ascending branch of the left colic artery is the primary supplying vessel (96.91%). Isoperistaltic transposition of the transverse colon is preferred (83.58%). Riolan's vascular arcade is not a major vessel of the colon. It can only be found in less than 10% of patients, and whether or not this arcade is complete cannot be used as criterion to judge colon blood supply. Animal experiments and clinical studies have confirmed the superiority of one-layer over two-layer anastomosis. The former is simpler, safer, and more reliable, with a lower incidence of anastomotic leak or stricture. Based on a comprehensive evaluation of the disease type, patient age, heart and lung functions, nutritional status, and accompanying diseases, three colon transposition routes are available (anterosternal subcutaneous tunnel, retrosternal tunnel, and esophageal bed passage). The advantages and disadvantages of each route are analyzed, and the left-middle-left retrosternal route is described. The indications for esophageal reconstruction with colon operation (ERC) were collected and verified, increasing the number of indications to seven categories of diseases. The main complications of ERC, i.e., colon segment necrosis, anastomotic leak, recurrent laryngeal nerve injury, and intestinal obstruction were systematically studied, and their causes and prevention are detailed. The number of patients is the highest in a single unit among all the published reports. The incidence of complications and deaths are the lowest. PMID- 10693251 TI - Photodynamic therapy. AB - Photodynamic therapy (PDT) is a treatment modality that utilizes a photosensitizing drug activated by laser-generated light and is proving effective for oncologic and nononcologic applications in the gastrointestinal tract. This article provides an overview of the most frequently used photosensitizers and clinical PDT studies in the upper gastrointestinal tract. In the future, the best indications for PDT in the esophagus will not comprise the palliative treatment of obstructing tumors but precancerous lesions such as Barrett's esophagus with or without dysplasia and early cancer. PDT might establish itself as a minimally invasive treatment alternative compared with surgery for high-grade dysplasia or early mucosal cancer of the esophagus. PMID- 10693252 TI - Endoscopic mucosectomy: an alternative treatment for superficial esophageal cancer. AB - Recent trends in the management of superficial esophageal cancer consist of improved detection, pretherapeutic staging and reliable criteria for curative endoscopic therapy. The endoscopic treatment is legitimate when the cancer is at an early stage, intra-epithelial or microinvasive (m1 or m2) and N0. Submucosal cancer should not be treated with a curative intent by endotherapy. Concerning squamous cell cancer, the oriental and occidental pathologists include high-grade dysplasia in the same group as intramucosal cancer. The distinction is however maintained for adenocarcinoma in the Barrett's esophagus. Indications of endoscopic rather than surgical treatment rely on: (1) the small size of the tumor (not more than 2 cm in diameter); (2) the endoscopic morphology in the type 0 of the Japanese classification with the flat subtypes IIa and IIb rather than type IIc--there is high risk of submucosal invasion for the polypoid (type I) or ulcerated superficial cancer (type III); and (3) the endoscopic ultrasound staging, with confirmed integrity of the hyperechoic submucosal layer. The high frequency (20 MHz) miniprobe is preferred to the standard (7.5 MHz) instrument. The elective procedure for tumor eradication is endoscopic mucosectomy. The technique is associated with a 6.8% risk of severe complications (hemorrhage or perforation) and a recurrence rate of 3%-7%. The 5-year survival rate is similar to that of surgery (over 80%). In the small group of patients with superficial esophageal cancer (less than 10% of the disease) endoscopic treatment may now be proposed in about 30% of cases, surgery is preferred for submucosal cancer and for neoplasia with a large surface. Areas of high-grade dysplasia in the Barrett's esophagus offer a new and increasing sector of indications. The concurrent endoscopic procedure of destruction--photodynamic therapy--is preferred for the destruction of lesions with poorly delineated limits. PMID- 10693253 TI - Quality of life in patients with oesophageal cancer. AB - There is a growing interest in assessing quality of life in patients with oesophageal cancer because it provides detailed information of the patients' perception of the benefits or harms of treatment. Yet few studies have prospectively measured quality of life using validated appropriate instruments. There are now several questionnaires for patients with cancer, although these are not sufficiently sensitive to small but clinically important changes in quality of life. It is therefore recommended that a disease-specific module is used in conjunction with generic measures. The European Organisation into Research and Treatment of Cancer (EORTC) QLQ-OES24 is currently completing an international validation study. It is used with the EORTC QLQ-C30 core instrument and is designed for patients undergoing potentially curative treatment or palliation of malignant dysphagia. Studies that have assessed quality of life after oesophagectomy have generally found that survivors do regain their former health. Little is known about the effect of neoadjuvant chemoradiation on patients' quality of life. Following endoscopic palliation of dysphagia, quality of life can be maintained and improvement of swallowing is seen. A validated appropriate assessment of quality of life should be included in future palliative trials and in studies of new treatments which may marginally influence survival but cause significant side effects. PMID- 10693254 TI - [How to examine a patient with Parkinson disease]. PMID- 10693255 TI - [Anti-neuronal antibodies and central nervous system diseases: contribution to diagnosis and pathophysiology]. AB - Antibodies against antigens found in the central nervous system have been evidenced in several neurological diseases. The most well-known are associated with paraneoplastic neurological diseases (Anti-Hu, Yo, Ri amphiphysin, Tr, CV2 and Ta antibodies). Some of these antibodies are specific for certain types of cancer or neurological syndromes and are highly useful diagnostic tools for the clinician. They have contributed to the hypothesis that these paraneoplastic neurological syndromes involve autoimmune cross reactions between tumoral and nervous system antigens. They are however most unlikely pathogenic on their own. Anti voltage-dependent calcium channel antibodies associated with Lambert-Eaton syndrome which is paraneoplastic in only 60 p. 100 of the cases are also observed in cases of paraneoplastic cerebellar atrophy. Anti-GAD antibodies are seen in non-paraneoplastic stiff man syndrome and in certain progressive cerebellar atrophies. Antibodies reacting with different glutamate receptors are detected in different neurological diseases including Rasmussen's encephalitis. Finally, antibodies are described in diverse conditions such as amyotrophic lateral sclerosis, Sydeham chorea or Gilles de la Tourette syndrome. The significance of the antibodies observed outside the context of paraneoplastic syndromes is not well understood, but the anti-GAD antibodies associated with progressive cerebellar disorders and autoimmune polyendocrinopathies could be an expression of the autoimmune nature of certain neurological degenerative processes affecting the central nervous system. PMID- 10693256 TI - [True neurological thoracic outlet syndrome]. AB - The thoracic outlet syndrome (TOS) encompasses various clinical entities affecting the neurovascular bundle crossing the thoracic outlet. Unfortunately, this term often proves to be confusing because many of these entities have little in common beyond their known or presumed lesion site. Neurogenic TOS (true TOS) is caused by compression of the lower trunk in the brachial plexus, the cervical ribs or fibrous band. This syndrome is extremely rare. We consider that this neurological form of TOS is a clearly defined neurological syndrome. We report 10 patients with true TOS. All were females. Stating the onset was difficult because symptoms were progressive and insidious. Pain was the most frequently reported symptom. Sensory deficit was slight or absent. All patients showed unilateral severe atrophy of the thenar muscles. Wasting and weakness developed later. A reduced amplitude of ulnar and median compound muscle action potential associated with a normal amplitude of median sensory nerve action and a reduced amplitude of ulnar sensory nerve action potential were indicative of a chronic axon loss in the lower trunk of the brachial plexus. In all cases, we performed medial antebrachial cutaneous sensory nerve action potential, a C8-T1 innervated nerve. The absence of the medial antebrachial cutaneous sensory nerve action potential in 9 patients and a reduction in amplitude of 50 p. 100 compared to the unaffected side in the other patient, indicated the diagnostic value of this easy and reproductible test. It confirmed a C8-T1 post-ganglionic radicular lesion or a lower brachial plexus neuropathy. Radiography showed a rudimentary bilateral cervical rib or an elongated C7 transverse process in all cases. Surgery was performed in the affected side in 7 patients and in each case the lower part of the brachial plexus was found to be stretched and angulated over a fibrous band, which was removed. Pain was relieved after 1 to 4 weeks. A minimal motor improvement was observed after one year. Electrophysiological results were unchanged. PMID- 10693257 TI - [Case study of 199 patients with multiple sclerosis: the use of EDMUS program]. AB - European Database Multiple Sclerosis (EDMUS) is a standardized tool which allows the collection of clinical, biological, radiological and therapeutic data on multiple sclerosis (MS). A descriptive analysis of 199 patients was done using EDMUS (version 2.2). Most data agreed with previous published results, except a low 0.4 sex ratio. At the beginning of MS, 42 p. 100 of MS patients developed pyramidal and sensory involvements and 14 p. 100 retrobulbar optic neuritis. Optic neuritis was the most frequent initial symptom of relapsing-remitting MS (25.9 p. 100) but not in the chronic progressive form (5.5 p, 100; p < 0.01). Surprisingly, relapsing-remitting form occurred more frequently whereas the secondary progressive form was diagnosed less. The concordances between the three disability scales, EGS (EDMUS Grading Scale), EDSS (Expanded Disability Status Score) and AI (Hauser Ambulation Index) were correct. In clinical practice, EDMUS is user-friendly and rapid. The lack of radiological data and the necessary quantification of only one disability scale, EGS may be too restrictive. The analysis of most of clinical data is possible by using integrated selection within the software, but sometimes needs more complex procedures. PMID- 10693258 TI - [Evaluation of motor speech function in the diagnosis of various forms of dysarthria]. AB - Perceptual analysis is not sufficient enough to identify specific dysarthria types. In order to improve the discrimination between dysarthria types, we developed a standardized evaluation of different functions controlling speech motor performances. This was applied to 90 patients suffering from hypokinetic, spastic or ataxic dysarthria and 15 control subjects. A discriminate analysis showed that 71.4 p. 100 of the cases were correctly classified. This model was validated within a new group of 21 patients and showed that the less severe dysarthric parkinsonian patients were difficult to distinguish from control subjects. The discriminate analysis was then used for 20 patients with atypical parkinsonism. Seven patients with progressive supranuclear palsy were considered to have hypokinetic dysarthria. The 6 patients with multisystem atrophy and 7 with corticobasal degeneration were classified among the 3 dysarthric types. We suggest that motor speech evaluation may contribute to differential diagnosis within groups of patients suffering from atypical parkinsonism. PMID- 10693259 TI - [Chemotherapy of malignant inoperable gliomas. The association of fotemustine cisplatine-etoposide as neoadjuvants]. AB - Efficiency of chemotherapy (CT) on non removable HGG has not been proven and neoadjuvant brain irradiation (RT) following biopsy is the standard treatment. We aimed to define whether combination of polychemotherapy and radiotherapy is synergistic in non removable HGG. It has been proven that F, CDDP and VP16 can reach therapeutic levels in brain after intravenous standard dose injections. The aim of this study was to assess that (i) neoadjuvant CT is safe; (ii) feasibility and efficacions of F (100 mg/m2.d1)/CDDP (100 mg/m2.d1-3 TD)/VP16 (75 mg/m2.d1-3) q21-28d regimen; (iii) Delayed RT is not unsafe: RT was performed when tumor progression or toxicity appeared. This study included 16 patients with symptomatic non removable HGG. Two of them had anaplastic gliomas and 14 glioblastomas multiforme. None of them had a prior chemotherapy regimen. Objective response was evaluated with CT scan or MRI during chemotherapy. Toxicity was moderate and mainly hematological (grade III-IV thrombopenia = 10/67 cycles; leukopenia = 13/67). Objective response rates were 5/16 (31 p. 100) (CR = 1; PR = 4; Median duration of response: 20 weeks). Median survival was 55 weeks in the 14 grade IV patients. Three/16 patients are still alived with respectively 22, 30, 40 months survival: These results confirm the neoadjuvant chemotherapy efficacy. It may be a useful tool before RT for non removable HGG. PMID- 10693260 TI - [Periodic hypokalemic paralysis as the manifestation of Graves' disease: clinical and electrophysiological study]. AB - A 30-year-old Vietnamese man experienced severe weakness of four limbs revealing hypokalaemia. Tachycardia and goiter allowed to diagnose a Graves' disease. Thyrotoxic hypokaliemic periodic paralysis is a rare complication of hyperthyroidism in western populations. Electromyogram performed after resolution of the paralysis episodes revealed marked abnormalities of the muscular excitability. Duration of compound muscular action potential was markedly increased after a brief exercise. This increase might indicate a delayed repolarization of muscle fibres. PMID- 10693261 TI - [Polyradiculoneuropathy in an adult with primitive hyperoxaluria]. AB - From the age of 31 a patient began to suffer from recurrent calcium oxalate urolithiasis. Liver biopsy showed a decrease in catalytic activity of the hepatic peroxisomal enzyme alanine: glyoxilate aminotransferase (AGT), which was mistargeted from peroxisomes to mitochondria. The genetic analysis revealed a mutation of the AGT gene. At age 47 he developed end-stage renal failure and underwent hemodialysis. After 12 months of hemodialysis he presented a rapidly declining clinical condition, a decrease of the residual renal function, a livedo reticularis with painful of extremities, and shortly thereafter a general weakness, which predominated on lower limbs. Apart from renal failure, routine biological examination and CSF were normal. Nerve conduction studies and electromyography supported the diagnosis of polyradiculoneuropathy. Pathological studies revealed mixed demyelinating-axonal lesions and deposits of calcium oxalate crystals within the media and the intima of epineural arterioles. A combined liver-kidney transplant was rapidly performed. The patient's condition improved in a few months and motor signs completely disappeared. PMID- 10693262 TI - [Neurological features after consumption of a variety of neo-caledonian shellfish]. AB - A few days after a seafood meal a patient suffered ataxia and stupor. His examination revealed a confused patient with cerebellar signs and ocular disturbances (hypotropia). Blood results, cerebrospinal fluid and brain CT scan were unremarkable. The patient developed a septic shock and died 4 weeks after admission. No necropsy was performed. Questioning his family confirmed that he had eaten a shellfish meal a few hours before onset of the digestive signs. Trocas (Tectus pyramis) were definitely identified. The clinical picture strongly suggested a seafood poisoning, namely ciguatera. However, no toxicologic assay was performed. To our knowledge, this poisoning has never been reported with trocas. Nevertheless, the feeding habits of trocas would suggest similarity with ciguatera poisoning. PMID- 10693263 TI - [Cerebral ischemia in the course of infectious endocarditis: a signal and a marker of progression]. PMID- 10693264 TI - [The opsoclonus-myoclonus syndrome]. PMID- 10693265 TI - [Electrodiagnosis of neuromuscular junction diseases]. PMID- 10693266 TI - [The first consensus conference in alcohology: objectives, indications and modalities for weaning the alcohol dependent patient]. PMID- 10693267 TI - [The dominant motivation in the mechanisms of the conditioned reflex]. AB - It is substantiated that the mechanisms of dominant motivations play an essential role in conditioning. It is shown that motivations change convergent and chemical characteristics of single neurons of different brain structures and, especially, their sensitivity to corresponding reinforcing stimuli. As a result, motivation plays a role of an initial "canvas", against the background of which molecular "engrams of reinforcement" are built. The processes of interaction between the dominant motivation and reinforcement are mainly addressed to the apparatus of the action result acceptor. It is shown that dominant motivations participate not only in construction of molecular reinforcement engrams but also in their forestalling retrieval. PMID- 10693268 TI - I. P. Pavlov and the development of neuroscience. AB - Ivan Petrovitch Pavlov significantly changed and developed our knowledge of the brain functions and of the behaviour by his fundamental experimental and theoretical work on the physiology and pathophysiology of the higher nervous activity. He was one of the scientists who prepared the development of neuroscience in our century. During the Pavlovian Conference, 1950 in Moscow Stalin and the Communist Party tried to dogmatize his and his pupil's fundamental theories. But his pupils continued to develop Pavlovian ideas in open discussions with representatives of other schools in a very creative way, opening the doors for a system approach to understand the integrative functional systems of brain and behavior. Pavlov emphasized the high plasticity of the central nervous system. He investigated the complex functional systems within the brain and between the organism and its environment, and the designed models for pathology of the higher nervous activity. During his last years, Pavlov freed himself from the strong deterministic view and characterized the organism and its environment as a self-organizing system. PMID- 10693269 TI - [I. P. Pavlov's theory on higher nervous activity: the landmarks and trends in its development]. AB - The theory of higher nervous activity created in Pavlovian time is compared with the present-day state of the theory developing due to appearance of new methods, techniques, facts, and concepts. Three principles of Pavlovian theory: determinism; analysis and synthesis; structural approach, as well s types of conditioned reflexes and techniques, types of higher nervous activity, and inhibition problems are discussed. The theory of higher nervous activity is schematically depicted as a tree, some branches of which are presented by facts and concepts obtained and introduced by I.P. Pavlov and his followers during his life, the others are formed by new facts and concepts advancing the theory. What is obsolete in the theory, what are the most prominent tendencies of its development and its new branches are discussed. PMID- 10693270 TI - [A physiologist's dialog with a psychologist]. AB - The famous polemics between I. P. Pavlov and K. S. Lashley in the pages of the "Psychological Review" in the thirties is discussed in this paper. The particular significance of this polemics for development of contemporary neuroscience is underlined, since the essentially novel concept of the brain activity mechanisms has been advanced and intelligence nature has been advanced on the basis of the synthesis of the two opposing theories, the localizationalism and equipotentialism. PMID- 10693271 TI - [Generators of the rhythmic alpha activity in the human EEG]. AB - An attempt to localize brain mechanisms of the rhythmic activity in the alpha rhythm range was made using the equivalent dipole model. It is known that light flickering stimuli with the frequency close to that of the individual alpha rhythm induce an increase in its spectral power ("photic-driving" phenomenon). It was shown that the activity of the neuronal structures generating the alpha rhythm can be identified by specific frequency of the light stimulation and localized by means of construction of dipole models. Two sources of the alpha rhythm in the narrow-frequency bands with the maximal resonance responses in the frequencies of 10.1 and 10.5 Hz were localized in the thalamic structures. PMID- 10693272 TI - [The characteristics of the electrodermal activity during changes in the level of human wakefulness]. AB - The wake-drowsiness transition during the performance of a monotonous psychomotor test was studied using polygraphic recording, with particular emphasis on changes in the electrodermal activity (EDA) and occurrence of electrodermal reactions (EDR). It was found that decrease in wakefulness was accompanied by a drop of the EDA. The EDR gradually disappeared for several minutes and did not reappear without activation; their dependence on sex and individual features of a subject significantly increased. It is suggested that discrepancies in the experimental results of psychophysiological studies with EDA may be explained by an insufficient control of the subject's alertness throughout the experiment. Examples of such discrepancies are given: disagreement of sex differences in the EDA, differentiation between EDA-labile and EDA-stable persons, on a possibility of recording the "emotional" EDA from different parts of human body etc. The ways of reproducibility of the results are discussed including the recording of several wake-drowsiness transitions and better recognition of the EDR. PMID- 10693273 TI - [The emotional assessment by test subjects of short musical fragments]. AB - Emotional estimation of 96 short (2-5 s) musical fragments was studied in 6 healthy subjects. Fragments were scored on a 5-point scale. Statistically significant difference in estimations of specific musical fragments (positive, negative, or indifferent) was found. PMID- 10693274 TI - [A clinical and neuropsychological analysis of mental functions in 6- to 7-year old children]. AB - The neuropsychological analysis of the state of higher mental functions (HMF) in children aged 6-7-years entering the first class of the school providing general education showed that the HMF of 52-73% of the future schoolchildren are underdeveloped with respect to their age. The underdevelopment of the emotional volitional sphere, of the dynamic organization of object activity, audio-speech memory, sphere of images-representations, visually-image and verbally-logical forms of thinking has been revealed. The experimental data suggest the absence of the primary (structural) derangements of the studied HMF in the examined children and dependence of the extent of their formation on the extent of readiness of the voluntary organization of activity. Insufficient formation of the HMF with respect to age is manifested in the lag of the formation of the internal structure of the psychic process, i.e., the absence of interiorization of the psychic processes and formation of the internal mental act. The neuropsychological assessment of underdevelopment of the HMF in reference to age suggests an insufficient age-related maturity of subcortical structures and subcortical corticopetal pathways as well as a dysfunction of the frontal neocortex and a secondary dysfunction of the posterofrontal and TPO areas, non auditory cortical temporal regions of the left hemisphere due to underdevelopment of connections of these areas with the frontal structures. PMID- 10693275 TI - [The physiological indices of emotional reactions in the clinical picture of local brain lesions]. AB - The psychophysiological correlates of recognition and reproduction of emotional states were studied in patients with differently localized lesions of the brain cortex. It was shown that the effect of different lesions are anisotropic. The cross and reciprocal interaction between the posteriofrontal and temporal regions of the left and right hemispheres was described. A relation was revealed between the emotion sign and different autonomic indices. A quantitative estimation of physiological reactions of healthy persons and patients with focal brain pathology was carried out. PMID- 10693276 TI - [A physiological analysis of dissociative learning against a background of physostigmine and pentobarbital]. AB - Physostigmine (0.7-0.8 mg/kg, i.p.) decreased and pentobarbital (13.4-14.6 mg/kg) increased the locomotor and emotional activity of rats in the "open field". Both drugs induced the reversible amnesia to a conditioned reaction in a double T-maze with positive (nutritional) reinforcement. These changes in behavioral activity were correlated with dissociated learning of rats after the injection of the drugs: physostigmine largely decreased the number of errors during learning as compared with pentobarbital. However, in both cases rats reached the learning criterion sooner than the control animals due to the shorter reaction latency (physostigmine) and increase in general motor activity (pentobarbital). PMID- 10693277 TI - [The transmission of coded information over neuronal systems exemplified by the motor rhythmic dominant]. AB - Functional organization of neurons in rabbit's sensorimotor cortex was studied before and within several days after formation of the rhythmical dominant focus. Functional reorganization of neurons in cortical microareas took place during actualization of the dominant. The number of functional interneuronal relations within neuronal pairs of a certain type could be increased in comparison with the control values and decreased within pairs of another type. As a result, the total percent of the interneuronal correlations in cortical microareas in the control animals and rabbits with the acting dominant was approximately equal. The total percent of correlations between neurons of the adjacent cortical areas during the actualization of the dominant was significantly higher than in the control due to increased number of correlations with participation of small and medium-sized neurons. A possibility of information circulation about the "stimulus image" in the closed chain of neurons was exemplified by the real micronetwork. The data suggest the reverberation of encoded information between adjacent microareas of the sensorimotor cortex within several days after application of the stimulus, which has formed the excitation focus. PMID- 10693278 TI - [The neurochemical mechanisms of the formation and consolidation of haloperidol induced catalepsy]. AB - In rat brain cortex, haloperidol initiates the long-term potentiation of K(+) induced Ca(2+)-dependent noradrenaline (NA) and dopamine (DA) secretion in vitro and in vivo. In both cases, the long-term potentiation is caused by the long-term increase in catecholamine content in the NA and DA terminals, as it has been shown in cortical tangential slices. Acute intraperitoneal haloperidol injection (2.5 mg/kg) evokes catalepsy and increases the content of NA and DA in the brain structures with localization of catecholamine receptors on terminals. This increase appears to be caused, predominantly, by modification of the terminal DA receptors, since only a trend to catecholamine increase is observed in the brain structures with a mixed type of NA and DA receptor localization (on somata and terminals). It is suggested that the long-term and diffuse action of haloperidol after its acute administration consists in the anxiogenic reaction and consolidation of catalepsy without an additional procedure of training and in the absence of unconditioned stimulus. PMID- 10693279 TI - [Generalized posttetanic changes in the excitatory postsynaptic and acetylcholine evoked currents in snail neurons]. AB - Heteroreceptor posttetanic changes in excitatory postsynaptic currents (EPSC) and inward currents evoked by the local iontophoretic application of acetylcholine (ACh) on the dorsal surface of PLa3 and PRa3 Helix lucorum neurons were studied. The following changes in the currents were revealed over the course of 1-1.5 h after tetanization. The rhythmical ACh application (0.5-1.0 cps, 10-40 s) evokes potentiation of the orthodromic EPSC. The tetanic orthodromic stimulation of one of the nerves (n. intestinalis, n. pallialis dexter, or n. pallialis sinister; 1 5 cps, 1-2 min) causes the potentiation of the ACh current and also heterosynaptic depression of the EPSC. It is concluded that activation of subsynaptic and nonsynaptic neurotransmitter chemoreceptors evokes the development of generalized posttetanic changes in neuronal responses. PMID- 10693280 TI - [Latent contextual inhibition of the cardiac component in the startle reaction to a sound stimulus in rats]. AB - Heart rate (HR), freezing score, and motor component were estimated during acoustic startle (ASR) habituation (two sessions with 24-hour interval, 10 trials in a session). It was shown that rats previously exposed to the experimental context (once for 5 min 24 h before training) demonstrated HR decrease in response to the first stimulus and tachycardia in response to the 4th-10th stimuli during the first session. The interstimulus HR declined from the 4th to the 6th trials. The same profile of cardiovascular response was observed in this group at the beginning of the second session with the following (to the 7th trial) habituation of tachycardia. These rats didn't demonstrate the intertrial HR decrease during the second session. Nonadapted rats responded by bradycardia to the 1st and 2nd trials of the first session. The response didn't change for tachycardia with continuation of stimuli presentation. Tachycardia appeared only in response to the 7th-10th trials of the second session and didn't habituate. The intertrial HR level decreased in this group only during the second session. The results are discussed in terms of contextual latent inhibition of the cardiac acoustic startle response. PMID- 10693281 TI - [The characteristics of the manifestation of hereditarily induced anxiety in male C57Bl/6J and CBA/Lac mice]. AB - Behavior of male mice of C57Bl/6J and CBA/Lac strains was tested in the elevated plus-maze and open field in order to estimate state anxiety in novel conditions. The cube and partition tests were used to reveal trait anxiety in the familiar conditions of the home cage. It is concluded that genetically defined state anxiety is more pronounced in CBA/Lac mice and trait anxiety in C57Bl/6J strain. PMID- 10693282 TI - [Anxiety and behavior in WAG/Rij strain rats with genetically induced absence attacks]. AB - Behavior of nonlinear rats and animals from Wistar and WAG/Rij (with inborn generalized absence epilepsy) strains was examined in the elevated plus-maze and the hole board. WAG/Rij rats demonstrated low exploratory behavior in both tests. In the elevated plus-maze, WAG/Rij rats were more balanced and more anxious than Wistar and nonlinear rats. Administration of ethosuximide completely eliminated spike-wave discharges but did not change behavioral interstrain differences. Since the spike-wave patterns develop in WAG/Rij at the age of 3 months, the behavior of young (2-moth-old) pups from different strains was compared and significant differences were revealed. Correlation between the genetically defined features (spike-wave discharges) and behavioral peculiarities in WAG/Rij rats is supposed. PMID- 10693283 TI - [The effect of alcoholization on the behavioral reactions of rats in an 8-arm radial maze]. AB - The influence of 20% alcohol consumption on training of low-active rats in 8-arm radial maze was studied. One group of animals was trained before and the other group after the alcoholization. All the animals acquired the conditioned reaction in the radial maze. However, the behavioral difference between the groups consisted in spatially-motor asymmetry. The rats trained before the alcohol consumption had less stereotyped behavior and more distinctly preferred to enter the maze arms at the angle of 45 degrees than the animals trained after the alcohol consumption. After the alcohol consumption, rats more frequently refused from behavioral task performance in comparison with the animals trained after the alcoholization. The influence of alcohol consumption of learning and memory in low-active rats is discussed. PMID- 10693284 TI - [The effect of different stages of the sex cycle on rat behavior in a plus maze]. AB - Anxiety and motor activity of female white rats in the elevated plus-maze were studied at different stages of the reproduction cycle (estrus, diestrus, pregnancy and lactation). The level of anxiety was lower, and that of locomotor and exploratory activity was higher during estrus and lactation than during diestrus and pregnancy. Exposure to chronic pain of threshold intensity did not induce behavioral changes in pregnant rats. There was no difference between the control and experimental animals in the level of plasma corticosterone. PMID- 10693285 TI - [The brightness components of the visual evoked potential to color stimuli in the rabbit]. AB - Two earliest components of visual evoked potentials (N85 and P130) which were related with substitution of stimuli for those identical in spectra but different in brightness were detected in rabbits. This finding suggests an analogy between the N85 and P139 in rabbits and N87 and P120 in humans. PMID- 10693286 TI - [The effect of scopolamine on the spatial organization of rat cortical potentials]. AB - The influence of scopolamine (1 mg/kg, i.p.) on the spatial organization of the neocortical electrical activity was studied in rats. A decrease in the spectral power and coherence of brain potentials in the range of the dominant theta-rhythm peak (6.00-7.25 Hz) and their increase in the adjacent low-frequency band were observed. Both indices were decreased in the wide beta band (19.00-30.00 Hz). The described changes took place over the whole areas of the right hemisphere and parieto-temporal region of the left hemisphere. The obtained results are discussed with respect to the role of the cholinergic brain system in the higher nervous activity. PMID- 10693287 TI - [Changes in the direct and interhemispheric responses of the pyramidal tract after tetanization of the cortex and lateral hypothalamus]. AB - Changes in pyramidal tract response after unilateral neocortical and lateral hypothalamic tetanization were analyzed in unanesthetized and nonimmobilized rabbits. Membrane and synaptic modifications were revealed in intra- and interhemispheric connections. Changes in excitability of callosal collaterals of pyramidal tract neurons in contralateral hemisphere and changes in somatodendritic excitability in ipsilateral hemisphere could be oppositely directed. Plasticity of callosal connections may contribute to the interhemispheric asymmetry during learning. PMID- 10693288 TI - [The duration of the storage of the electrical characteristics of command neurons in the acquisition of long-term sensitization in the snail]. AB - The retention of the long-term sensitization (LTS) of defensive reflex and dynamics of change in electric characteristics (membrane potential (Vm) and action potential generation threshold (Vt)) of command neurons of defensive reflex was studied in a snail during behavioral tests. The membrane mechanisms were analyzed by measuring electrical characteristics of the LPa3, RPa3, LPa2, and RPa2 command neurons on the 1st, 4th, 7th, 10th, and 14th days after the LTS formation and 1 month later. The membrane potential and threshold potential in sensitized snails (-54.1 +/- 2.0 and 24.5 +/- 1.4 microV, respectively) were significantly (p < 0.001) decreased in comparison with the control animals (-60.9 +/- 0.8 and 19.9 +/- 0.6 microV respectively). These changes retained within 14 days after the LTS formation. The results suggest the long-term retention of the increased excitability of command neurons. A month after the LTS formation, the duration of the defensive reflex returned to the initial level and the electric characteristics of command neurons did not significantly differ from the control (-61.1 +/- 2.0 and 19.3 +/- 1.4 microV, respectively). PMID- 10693289 TI - [The interrelations of I. P. Pavlov and M. V. Nesterov]. PMID- 10693290 TI - Application of CT imaging for dental implant simulation. AB - Accurate diagnosis and exact treatment planning are very important for successful implant treatment. Pretreatment examination and simulations using CT can be especially effective information sources, and diagnosis by CT imaging before treatment substantially enhances the security and safety of the treatment plan. New interactive CT software (SIM/Plant) which enables diagnosis of bone morphology and quality, and also implant simulation on a personal computer through multiplanar reformation of CT images, has been developed. Stereolithography models (SLMs) are reproduced anatomical morphological models of an individual patient's bone structure from the information obtained by the CT scan, and is fabricated out of light cure resin. In this article, SLMs and SIM/Plant for simulation of various examinations and diagnosis incorporating CT information are described, and their features are introduced. PMID- 10693291 TI - IL-6 levels in gingival crevicular fluid (GCF) from patients with non-insulin dependent diabetes mellitus (NIDDM), adult periodontitis and healthy subjects. AB - Cytokines play an important role in the pathology associated with chronic inflammatory diseases. One of these cytokines, interleukin 6 (IL-6) is a major mediator of the host response to tissue injury, infection and bone resorption. In the present study, gingival crevicular fluid (GCF) level of IL-6 was determined in patients with non-insulin dependent diabetes mellitus (NIDDM) with periodontitis, adult periodontitis, and healthy controls by use of an enzyme linked immunosorbent assay (ELISA). Twenty-four NIDDM patients with periodontitis, twenty-four adult periodontitis and twenty-four healthy controls were selected for the study. GCF sampling was performed on the vestibular aspects of maxillary incisors and canine teeth. Plaque index (PI), gingival index (GI), gingival bleeding time index (GBTI), probing depth (PD) and probing attachment levels (PAL) were recorded from each sampling area and also the entire dentition. NIDDM and adult periodontitis patients had numerous sites with radiographic evidence of alveolar bone resorption, loss of attachment and pocket depth greater than 3 mm. The mean GCF IL-6 level was 2.43 +/- 0.97 ng/ml in NIDDM patients, 1.31 +/- 0.92 ng/ml in adult periodontitis and 0.62 +/- 0.58 ng/ml in healthy subjects, respectively (p < 0.05). GCF IL-6 levels were markedly higher in NIDDM and adult periodontitis groups compared to the healthy controls. No correlation was found between GCF IL-6 levels and all clinical parameters. These findings suggested that GCF IL-6 levels were significantly higher in the area of inflammation and periodontal destruction locally. The high IL-6 levels in NIDDM patients might be due to different microbial flora in periodontal pockets and altered immune system. Future studies are needed to evaluate the complex interaction among IL-6 GCF levels, host response and local microbial environment in the NIDDM patients. PMID- 10693292 TI - The phenomenon of salivary protein adsorption onto Streptococcus mitis ATCC 903 cells. AB - It is thought that salivary proteins on bacterial cells have some influence on the adhesion of oral bacteria onto the surface of oral tissues and on bacterial aggregation. Initially, this study sought to examine the phenomenon of salivary protein adsorption to the surface of Streptococcus mitis ATCC 903 using 3H labeled salivary proteins. We investigated the effects of hydrophobic level and lectin-ligand binding on adsorption of salivary proteins, and also the influence of cell surface components on adsorption. The results showed that the adsorption of salivary proteins was clearly reduced by the presence of Tween 20, LiCl, NaSCN, Hexadecane and some sugars. The adsorption was also affected by the pH level, and protease treatment or heat treatment of cells also decreased the volume of adsorbed proteins. Although the adsorption of proteins onto heat treated cells was dramatically reduced by the presence of LiCl and NaSCN, the presence of sugars had little influence on this adsorption. These findings suggest that the main adsorption of salivary proteins is due to hydrophobic factors, and the heat-sensitive surface components of the cells had some relation to lectin-ligand binding. Therefore, it appears important to study the adsorption of salivary proteins onto cells since the salivary proteins on bacterial cells play an important role in their adherence to the saliva-coated oral tissues and bacterial aggregation. PMID- 10693293 TI - Clinicopathological studies of odontoma in 47 patients. AB - A 14-year retrospective study was performed on 47 odontomas from the files of the 1st Department of Oral and Maxillofacial Surgery at Nihon University School of Dentistry. Fifty-seven percent of the patients were male and 42.6% were female. The age distribution was 8 to 48 years with a mean age of 22 +/- 9.0 years. There were no particular symptoms associated with the odontomas, and 63.8% of our patients had no symptoms. However, 12 patients complained of swelling and 9 of pain. The tumor was found in the maxilla in 42.6% and in the mandible in 57.4%. According to the WHO histological type classification, 53.2% of the tumors were classified as compound odontoma and 46.8% as complex odontoma. The size of the tumor ranged from 5 mm to 42 mm in diameter. The average complex odontoma was much bigger than the average compound odontoma. Ghost cells were found 11 cases in our series. In addition, odontogenic epithelium was found in 16 cases. Twenty seven patients had impacted teeth in association with odontoma and 24 of the 27 teeth were removed at the time of surgical enucleation of the tumor, while 3 cases were treated by orthodontically assisted eruption. There was no recurrence in any of the studied cases. PMID- 10693294 TI - Histological assessment of root cementum at periodontally healthy and diseased human teeth. AB - This study aimed to investigate and compare the lipid and polysaccharide content of the cemental surfaces of healthy and periodontally-involved teeth. Thirty periodontally-involved single-rooted teeth from fifteen patients with localized juvenile, adult and rapidly progressive periodontitis were included in the experimental group and 5 healthy teeth were assessed in the control group. Frozen serial sections were obtained and stained with hematoxylin-eosin for morphological assessment. Oil-Red-O and Alcian Blue-Periodic Acid Schiff stains were used to evaluate the presence of lipids, neutral and acidic polysaccharides using light microscopy. It was found that with hematoxylin-eosin staining in the experimental group, both the involved and uninvolved cementum surfaces of teeth, which belong to all periodontitis groups, showed generally irregular surfaces that contain some resorption areas. Alcian Blue-Periodic Acid Schiff positive staining was observed only superficially and at the areas associated with microbial dental plaque. However, Oil-Red-O staining was positive only superficially at 5 teeth that belonged to localized juvenile and rapidly progressive periodontitis groups. Apparent lipopolysaccharide staining into cementum was not seen in any of the diseased teeth. The results presented here suggest that endotoxin was only localized in superficial layers and associated with only microbial colonization. PMID- 10693295 TI - Treatment of ligature-induced peri-implantitis defects by regenerative procedures: a clinical study in dogs. AB - We carried out a clinical evaluation of the hard tissue fill following treatment of ligature-induced peri-implantitis in dogs. Four dogs were used and their mandibular premolars (P2, P3 and P4) were removed. After 3 months of healing, two titanium implants were placed on each side of the mandible. After 3 months, the abutment connection was performed, and experimental peri-implantitis was induced by placement of cotton ligatures in a submarginal position. The ligatures and abutments were removed after one month, and the peri-implant bone defects were assigned randomly to one of the treatments: debridement (control), debridement plus guided bone regeneration (GBR), debridement plus mineralized bone graft (BG), and debridement plus guided bone regeneration associated with a mineralized bone graft (GBR + BG). Clinical measurements of the peri-implant bone defects before and 5 months after treatment revealed no statistically significant differences between the defects treated by GBR, BG and GBR + BG. These 3 treatment methods provided more hard tissue fill than debridement alone (p < 0.05). Thus, it can be concluded that GBR, BG or a combination of the two techniques can enhance the hard tissue fill in defects caused by peri-implantitis in dogs. PMID- 10693296 TI - Beta-lactamases producing anaerobic bacteria in dentoalveolar abscesses. AB - Pus samples of 30 patients with closed dentoalveolar abscesses who had not received antimicrobial therapy for at last two months were screened for the presence of beta-lactamase-producing anaerobic bacteria. From these 30 pus samples, a total of 112 bacterial strains were isolated; 83 of them were strict anaerobes and 29 were aerobes. beta-lactamases activity of the selected anaerobic bacteria was tested after identification of the isolates and was detected in 5 of the total 83 (6%) strict anaerobic isolates, whereas these 5 strains were isolated in 4 of the 30 (13.3%) pus samples. The species with beta-lactamase activity were in the Prevotella intermedia (4 from 14 isolates) and the Fusobacterium nucleatum (1 from 9 isolates) groups. None of the gram-positive and the other gram-negative anaerobic strains were beta-lactamase positive. PMID- 10693297 TI - Evaluation of human tooth structure with the ultrasonic imaging technique. AB - In recent years, ultrasonic waves have been an interesting subject for studies due to their wide range of applications in medical diagnoses. In this study, the acoustic properties of the structure of human teeth was determined with the ultrasonic imaging technique. This study may offer some fundamental findings related to the clinical application of the ultrasonic imaging technique in the further development of a virtual system for dental education and research. Twenty freshly-extracted permanent human teeth (10 molars and 10 premolars) were used to investigate their acoustic velocity and impedance by the ultrasonic image analyzing system with a high-resolution focusing probe. Additionally, the relationship between the acoustic properties and the hardness of the teeth was evaluated. It was found that the acoustic properties of the human teeth were influenced by factors related to their structure, such as degree of calcification, distribution of dentinal tubules, and volume of the dentin matrix. The acoustic velocity and impedance showed an apparent correspondence to the hardness of tooth. This analyzing system provides visual information related to tooth structure that can easily quantitatively evaluate their acoustic properties. It is expected that this system will have a wide range of applications and be further developed for clinical uses. PMID- 10693299 TI - At the end of life: giving thanks and forgiving. PMID- 10693298 TI - Supernumerary teeth with eumorphism in the lower incisor region: a report of five cases and a review of the literature. AB - Five cases of supernumerary teeth with eumorphism in the lower incisor region of the permanent dentition are reported. The patients were two males and three females. One (a 31-year-old woman) of them had bilateral supernumerary teeth. The review of the English language literature yielded only one such bilateral supernumerary teeth in the lower incisor region of the permanent dentition was reported. PMID- 10693300 TI - What constitutes high-quality HIV/AIDS palliative care? AB - The study assessed the components of high-quality HIV palliative care using the multidimensional model of quality assessment developed by Maxwell (4). Data collection consisted of interviews with individual subjects and focus groups. The interviews took place in three London health authorities. Seven service users and 74 providers of health care and voluntary services were interviewed. Maxwell's model identifies the components of high-quality HIV palliative care as: competent, skilled practitioners (effectiveness); confidential, non discriminatory, and culturally sensitive care (acceptability); collaborative and coordinated care (efficiency); flexible and responsive care (access and relevance to need); and fair access for all clients (equity). Our results show that the six dimensions of quality appear to be inextricably linked from the client's point of view. Thus, service commissioners and providers need to examine all six of the dimensions when assessing the quality of HIV services. These findings may serve as a model for other palliative care services. PMID- 10693301 TI - Age is not the crucial factor in determining how the palliative care needs of people who die from cancer differ from those of people who die from other causes. AB - A belief that the hospice philosophy is particularly applicable to younger people may account in part for the continued focus of palliative care on cancer patients, as it has been argued that age is the crucial factor in determining how cancer and non-cancer patients differ. We conducted a secondary analysis of the data from a UK population-based retrospective survey, the Regional Study of Care for the Dying, to critically examine this proposition. The sample comprised 2062 cancer and 1471 non-cancer deaths. On average cancer patients were younger. However, at all ages non-cancer and cancer patients differed significantly with, for example, different patterns of dependency and symptomatology. The cause of death--rather than age--is therefore the principal difference between cancer and non-cancer patients. The debate within palliative care on whether and how to provide services for non-cancer patients must move beyond a focus on group differences such as age between these and cancer patients and focus instead on understanding the varying problems non-cancer patients experience, and addressing how best to organize palliative care services to meet the individual needs of these patients. PMID- 10693302 TI - Acupuncture for patients in hospital-based home care suffering from xerostomia. AB - A total of 20 patients (17 cancer patients) in late-stage palliative care reporting dryness of mouth and associated problems were treated for 5 weeks with 10 acupuncture treatments. Ten patients were also evaluated for speech problems (articulation) and dysphagia caused by xerostomia. Subjective assessments used Visual Analog Scales (VAS). Acupuncture had a dramatic effect on xerostomia and, subsequently, on dysphagia and articulation, with subjects showing definite improvement after 5 treatments. Release of neuropeptides that stimulate the salivary glands and increased blood flow are possible explanations for the effects. PMID- 10693303 TI - Symptoms as meaningful "family culture" symbols in palliative care. PMID- 10693304 TI - Terminal cancer syndrome: myth or reality? PMID- 10693305 TI - Are some palliative care delivery systems more effective and efficient than others? A systematic review of comparative studies. PMID- 10693306 TI - Predictors of family satisfaction with an Australian palliative home care service: a test of discrepancy theory. AB - Five interesting findings emerged from this study: Although study results demonstrate support for Porter's Discrepancy Theory, the most compelling outcome is the finding that family care perceptions may be the best predictor of family care satisfaction. Family members' age may be a predictor of family care satisfaction. Family functioning may be a useful clinical indicator to identify families who are less satisfied with care and in greater need of support. The length of time that clients receive the care service may alter family care satisfaction. Differences in findings reported in this study compared with Canadian results point to the need for cross-cultural research in this area. This research is the first Australian study to test discrepancy theory as a framework for understanding family care satisfaction in a home hospice context. Results from this study may assist health care providers to more sensitively address the care perceptions of families in this care setting and extend theory development research that is needed to guide palliative care practice with families. PMID- 10693307 TI - Practitioners' attitudes towards ethical issues at the end of life: is the UK actually more autonomy-minded than the US? PMID- 10693308 TI - Traditions associated with dying in the west of Scotland. PMID- 10693309 TI - Care for Chinese palliative patients. PMID- 10693310 TI - Increased awareness of osteopathic medicine is essential to the profession's survival. PMID- 10693311 TI - 'Unity' benefits extend beyond AOA public relations programs. PMID- 10693312 TI - In defense of Dr Kevorkian. PMID- 10693313 TI - Osteopathy an independent system co-extensive with the science and art of healing. 1901. PMID- 10693314 TI - Troponin I sensitivity and specificity for the diagnosis of acute myocardial infarction. AB - This article describes the sensitivity and specificity of troponin I when compared to creatine kinase-MB (CK-MB) and electrocardiography (ECG) for diagnosing acute myocardial infarction (AMI). Two different lower levels for defining positive results with troponin I were evaluated. A retrospective study of 153 patients who presented to the emergency department of a community hospital supplied the pool of patients for this study. Patients included in this study were those for whom a CK-MB was ordered. The majority of these patients were evaluated for chest pain or symptoms suggesting an acute cardiac event. Of the 153 patients studied, CK-MB results were positive in 91 (59%) patients; ECG revealed AMI in 72 (47%) patients. There were 103 (67%) patients who had either positive CK-MB or ECG results. Ninety (59%) patients had a troponin I level greater than 2.0 ng/mL, and 18 (12%) patients had a troponin I level between 0.6 and 2.0 ng/mL. Seven patients whose troponin I level was between 0.6 and 2.0 ng/mL had negative CK-MB and ECG results. Therefore, 11 patients with troponin I between 0.6 and 2.0 ng/mL had AMI. Five patients with positive troponin I results (> 2.0 ng/mL) had negative CK-MB and ECG results. When a troponin I level greater than 0.6 ng/mL was used as a positive value, compared to CK-MB and ECG using either time zero or time 6 hours, the sensitivity was 94% and specificity was 81%. When troponin I greater than 2.0 ng/mL was used to define a positive test, the sensitivity was 85% and specificity was 91% when compared to CK-MB and ECG. PMID- 10693315 TI - Role of antileukotriene agents in asthma therapy. AB - Leukotrienes are proinflammatory mediators with special significance in asthma. Released by numerous cell types, particularly after exposure to allergens, leukotrienes cause a potent contraction of bronchial smooth muscle, resulting in reduced airway caliber. Further, they cause plasma to leak from the vessels, resulting in edema, and enhance the secretion of mucus--both events that increase airway obstruction. Thus, leukotrienes have been a target of basic research in asthma. To date, a number of antileukotriene agents have been developed, and three are currently being used in clinical practice in the United States: zafirlukast and montelukast act by antagonizing the leukotriene receptor, and zileuton inhibits leukotriene synthesis. Studies have shown that these agents improve asthma symptoms, pulmonary function, and patient quality of life. Antileukotriene agents have generally been associated with a low incidence of side effects and good tolerability. Currently recommended for mild-to-moderate, persistent asthma, these agents have enabled patients to reduce their use of corticosteroids. PMID- 10693316 TI - Advances in forensic assessment and treatment: an overview and introduction to the special issue. PMID- 10693317 TI - The quality of forensic psychological assessments, reports, and testimony: acknowledging the gap between promise and practice. AB - During the past decade, the field of forensic psychological assessment entered a period of standard setting, reflected in the publication of specialty guidelines for practice and in the proliferation of educational opportunities, training programs, and credentialing and certification procedures for forensic examiners. Representing significant efforts to advance the quality of psychological assessments in legal contexts, these developments foreshadow the promise of forensic assessment. During this same time period, new evidence emerged regarding the quality of forensic practice. This article reviews this evidence and evaluates current practice against the promise of forensic assessment. Forensic reports appear to be of higher quality than those described by commentators in the 1970s and early 1980s; nevertheless, the level of practice falls far short of professional aspirations for the field. The review identifies significant areas of weakness that demand the attention of professional organizations, accrediting agencies, educators, lawmakers, practitioners, and consumers. PMID- 10693318 TI - Psychopathy (PCL-R) as a predictor of violent recidivism among criminal offenders with schizophrenia. AB - Hare's Psychopathy Checklist--Revised (PCL-R) was used to test the hypothesis that psychopathy predicts violent recidivism in a cohort subjected to forensic psychiatric investigation and consisting of male violent offenders with schizophrenia (N = 202). Psychopathy was assessed with retrospective file-based ratings. Mean follow-up time after detainment was 51 months. Twenty-two percent of the offenders had a PCL-R score > or = 26 (cutoff), and the base rate for violent recidivism (reconvictions) during follow-up was 21%. Survival analysis revealed that psychopathy was strongly associated to violent recidivism (log-rank = 17.71, df = 1, p < 0.0001). The area under the curve (AUC) of the receiver operating characteristics (ROC) of PCL-R total score to predict violent recidivism varied between different time frames from .64 to .75. Cox regression analyses revealed that other potential risk factors could not equally well or better explain violent recidivism in the cohort than psychopathy as measured by PCL-R. PMID- 10693319 TI - Development and validation of the Validity Indicator Profile. AB - The Validity Indicator Profile (VIP; Frederick, 1997) is a two-alternative forced choice (2AFC) procedure intended to identify when the results of cognitive and neuropsychological testing may be invalid because of malingering or other problematic response styles. The test consists of 100 problems that assess nonverbal abstraction capacity and 78 word-definition problems. The VIP attempts to establish whether an individual's performance in an assessment battery should be considered representative of his or her true overall capacities (valid or invalid). Performances classified as valid are classified as "compliant" and reflect a high effort to respond correctly. Performances classified as invalid are subclassified as "careless" (low effort to respond correctly), "irrelevant" (low effort to respond incorrectly), or "malingering" (high effort to respond incorrectly). The VIP development sample included 944 nonclinical participants and 104 adults undergoing neuropsychological evaluation. The cross-validation sample consisted of 152 nonclinical participants, 61 brain-injured adults, 49 individuals considered to be at risk for malingering, and 100 randomly generated VIP protocols. The nonverbal subtest of the VIP demonstrated an overall classification rate of 79.8%, with 73.5% sensitivity and 85.7% specificity. The verbal subtest of the VIP demonstrated an overall classification rate of 75.5%, with 67.3% sensitivity and 83.1% specificity. PMID- 10693320 TI - A classification tree approach to the development of actuarial violence risk assessment tools. AB - Since the 1970s, a wide body of research has suggested that the accuracy of clinical risk assessments of violence might be increased if clinicians used actuarial tools. Despite considerable progress in recent years in the development of such tools for violence risk assessment, they remain primarily research instruments, largely ignored in daily clinical practice. We argue that because most existing actuarial tools are based on a main effects regression approach, they do not adequately reflect the contingent nature of the clinical assessment processes. To enhance the use of actuarial violence risk assessment tools, we propose a classification tree rather than a main effects regression approach. In addition, we suggest that by employing two decision thresholds for identifying high- and low-risk cases--instead of the standard single threshold--the use of actuarial tools to make dichotomous risk classification decisions may be further enhanced. These claims are supported with empirical data from the MacArthur Violence Risk Assessment Study. PMID- 10693321 TI - The Spousal Assault Risk Assessment (SARA) Guide: reliability and validity in adult male offenders. AB - We evaluated the reliability and validity of judgments concerning risk for violence made using the Spousal Assault Risk Assessment Guide (SARA; Kropp, Hart, Webster, & Eaves, 1994, 1995, 1998). We analyzed SARA ratings in six samples of adult male offenders (total N = 2681). The distribution of ratings indicated that offenders were quite heterogeneous with respect to the presence of individual risk factors and to overall perceived risk. Structural analyses of the risk factors indicated moderate levels of internal consistency and item homogeneity. Interrater reliability was high for judgments concerning the presence of individual risk factors and for overall perceived risk. SARA ratings significantly discriminated between offenders with and without a history of spousal violence in one sample, and between recidivistic and nonrecidivistic spousal assaulters in another. Finally, SARA ratings showed good convergent and discriminant validity with respect to other measures related to risk for general and violent criminality. PMID- 10693322 TI - Improving risk assessments for sex offenders: a comparison of three actuarial scales. AB - The study compared the predictive accuracy of three sex offender risk-assessment measures: the RRASOR (Hanson, 1997), Thornton's SACJ-Min (Grubin, 1998), and a new scale, Static-99, created by combining the items from the RRASOR and SACJ Min. Predictive accuracy was tested using four diverse datasets drawn from Canada and the United Kingdom (total n = 1301). The RRASOR and the SACJ-Min showed roughly equivalent predictive accuracy, and the combination of the two scales was more accurate than either original scale. Static-99 showed moderate predictive accuracy for both sexual recidivism (r = 0.33, ROC area = 0.71) and violent (including sexual) recidivism (r = 0.32, ROC area = 0.69). The variation in the predictive accuracy of Static-99 across the four samples was no more than would be expected by chance. PMID- 10693323 TI - Expert approaches to communicating violence risk. AB - There has been virtually no empirical study of the way in which evaluating clinicians communicate their conclusions about the risk of violence toward others. Risk communication has become particularly important in recent years, serving as the link between empirical data from recent studies and the understanding and use of such data by evaluators and decision makers. The present study considered how psychologists and psychiatrists, identified as experts in violence risk assessment, responded to eight vignettes that systematically measured preferences for risk communication. The vignettes involved the presentation of the following factors in a 2 x 2 x 2 within-subjects design, counterbalanced for order: (1) risk model (prediction vs. management), (2) risk level (high vs. low risk of the individual being assessed), and (3) risk factors (the predominance of static vs. dynamic risk factors). A total of 71 individuals (41 psychologists, 2 sociologists, and 28 psychiatrists) responded to a survey mailed to 100 individuals, for a response rate of 71%. Participants were asked to rate the value of six forms of risk communication for each of the eight vignettes. There were few significant differences between the ratings assigned by psychologists and those assigned by psychiatrists. The most highly valued form of risk communication involved identifying risk factors applicable to the individual and specifying interventions to reduce risk. A repeated-measures multivariate analysis of variance yielded a main effect for risk level and an interaction between risk level and risk factors. The implications of these findings for research and practice are discussed. PMID- 10693324 TI - Distribution, density, and structure of muscle spindles in the vastus intermedius and the peroneus longus muscles of sheep. AB - Muscle spindles are not always distributed more in postural muscles with many slow-twitch-oxidative (SO) myofibers than in locomotory muscles with few SO myofibers. The purpose of present study was to examine the distribution, density, and structure of muscle spindles in the vastus intermedius muscle: an antigravity muscle and the peroneus longus muscle: a locomotory muscle in the sheep. Muscle spindles were reconstructed from serial sections at 300 microns intervals throughout the muscles. Myofiber types were classified into SO, fast-twitch oxidative-glycolytic, and fast-twitch-glycolytic myofibers by differences in histochemical reactivity. No significant difference in the density of muscle spindles (DMS) existed between the vastus intermedius (DMS: 5.3) and peroneus longus (DMS: 5.7) muscles. The muscle spindles were distributed more in the distal portion than in the proximal portion of the vastus intermedius muscle. The muscle spindles were distributed in the proximal and middle portion but hardly in the distal portion of the peroneus longus muscle. Muscle spindles were classified into simple, tandem, and compound muscle spindles. Most of the muscle spindles were the simple type. The differences in size of the muscle spindle and numbers of the intrafusal myofibers were not significant between the two muscles. The results show that the density and structure of the muscle spindles do not differ between the postural and locomotory muscles in the sheep. PMID- 10693325 TI - Macrophages in the hamster parathyroid gland: immunohistochemical and ultrastructural investigations. AB - The distribution and morphology of the parathyroid macrophages in golden hamsters from neonatal to senile periods were investigated using the monoclonal antibody to ED2 and electron microscopy. The results showed that definite ED2-positive cells were hardly detectable in the parathyroid gland of 1-day-old hamsters. A few ED2-positive cells could be identified in the parathyroid gland of 10-day-old hamsters. The ED2-positive cells were more densely and became conspicuous in 1-, 3-, and 12-month-old hamsters. The number of the cells seems to be increased with aging. Ultrastructurally, we did not find any macrophages in parathyroid glands of 1-day-old hamsters. In the 10-day-old hamster parathyroid gland, a few macrophages existed only in the interstitial tissues. In the parathyroid gland of 1-, 3-, and 12-month-old hamsters, many macrophages were found in the perivascular regions, some cells located among the parenchymal chief cells with no obvious vascular association. These cells showed some physical contacts with chief cells. These results suggest that the parathyroid macrophages exhibit dramatical changes in their distribution and morphology from neonatal to senile periods. PMID- 10693326 TI - An odontometrical difference in the mandibular molars of two laboratory strains of the musk shrew, Suncus murinus, derived from Bangladesh and Tokunoshima Island of Japan. AB - We investigated an odontometrical difference in the mandibular molars (M1, M2, and M3) of two laboratory strains of the musk shrew (Suncus murinus) originating in Bangladesh (BAN strain) and Tokunoshima Island of Japan (TKU strain). We used skulls from two strains of shrews that were maintained under identical laboratory conditions. Mesiodistal and buccolingual crown diameters in the trigonid and talonid of the mandibular molars were measured with a measuring microscope, calibrated to 0.001 mm. The crown proportion was expressed by the crown indices calculated from the measurements. Size reduction was analyzed quantitatively according to the reduction index. All crown dimensions were significantly larger in BAN shrews than in TKU shrews (P < 0.01). Sexual differences were noted in the talonid dimensions, while interstrain differences were clearly evident in the trigonid dimensions. The crown indices in M1 showed the least interstrain difference of the three molars. The crown indices showed that TKU shrews had relatively larger buccolingual diameters and talonid diameters than BAN shrews, and the reduction indices showed that TKU shrews had relatively larger M2 and M3 than BAN shrews. To extract the variance components of tooth shape, a principal component analysis was performed after the variables were standardized. After Varimax rotation, each factor was interpreted. The first three factors accounted for 79.9% of all variances. The first component represented the mesiodistal crown proportion of the trigonid-to-talonid crown component. The second and third components represented the relative size of buccolingual diameters in the distal molars for M1. The principal component scores showed that TKU shrews had relatively larger talonids and distal molars than BAN shrews. PMID- 10693327 TI - Sexual dimorphism of the human spinal cord in the aging process. AB - There have been few morphometric studies on age-related changes in the human spinal cord. The purpose of the present study was to determine the existence of sexual dimorphism of the spinal cord between males and females during the aging process. Spinal cords were removed from cadaver specimens, 26 males and 22 females for anatomic practice, the age at death ranged from 41 to 97 years for males (average, 71.5 years) and from 59 to 92 years for females (average, 76.6 years). Spinal cord segments were embedded in celloidin after secondary fixation and dehydration. Sections were stained with the Luxol fast blue-periodic acid Schiff-hematoxylin and Kluver-Barrera methods. Morphometric analysis was performed with an electronic optical planimeter and a computer. Each section was enlarged 13.5 times to take a picture. The areas of the transverse section white matter and gray matter of the spinal cord at segments C5 and L3 were measured. Although there was no correlation between the total transverse area of the spinal cord and age either in males or females, we noticed that the area of the gray matter decreased faster in males than in females; while the area of the white matter decreased faster in females than in males. The area ratio of the white matter to the whole segment area of the spinal cord (W/T) at level C5 is larger in males than that in females. Our results suggest that there could be a difference between males and females in changes in the white and gray matters of the spinal cord due to aging. PMID- 10693328 TI - Morphology of the dorsal lingual papillae in the blackbuck, Antilope cervicapra. AB - The dorsal lingual surface of a blackbuck (Antilope cervicapra) was examined by scanning electron microscopy (SEM). The tongue was about 125 mm in length. There were about 30 vallate papillae on both sides. Filiform, conical, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue, excepted for the lingual torus where conical papillae were present. The fugiform papillae were present rounded bodies, and more densely distributed on the tip and ventral surface of ligual apex. No foliate papillae were seen on the dorsal surface. The vallate papillae were located on both sides of the midline in the caudal part. Each papilla was surrounded by a groove. These findings indicate that the tongue of the blackbuck is similar to that of the formosan and japanese serow. PMID- 10693329 TI - The variations of the subclavian artery and its branches. AB - This study reports important variations in branches of the subclavian artery in a singular cadaver. The origin of the left vertebral artery was from the aortic arch. On the right side, no thyrocervical trunk was found. The two branches which normally originate from the thyrocervical trunk had a different origin. The transverse cervical artery arose directly from the subclavian artery and suprascapular artery originated from the internal thoracic artery. This variation provides a short route for posterior scapular anastomoses. An awareness of this rare variation is important because this area is used for diagnostic and surgical procedures. PMID- 10693330 TI - A radiological method on the classification of human mandibular condyles. AB - A radiological quantitative method applied in the classification of 210 mandibular condyles from 105 male and female subjects, aged 18-62 with no temporomandibular disorders has been taken up in this study. Through coronal sections, types of condyles were first determined by computer tomography (CT) per anterio-posterior aspects and then divided into four main groups described as flat, convex, angled and round. Then, using the parameters, every condyle type was divided into three subgroups. Finally, the percentage of each group has been estimated as follows: flat, 14.3%; convex, 35.2%; angled, 35.7% and round, 14.8%. When the condylar type was compared with sex, it was observed that the angled type (39.6%) in males and the convex type (40.3%) in females were higher than the other types. In the analysis of the relationship between age and condylar types, there appeared to be a trend toward an increase in the incidence of flat and a decrease in the incidence of convex with an increase in the age of males. In addition, following the determination of bilateral symmetry and asymmetry the bilateral asymmetry was determined to be higher in both sexes (56.2%). PMID- 10693331 TI - Variations of nerves located in deep gluteal region. AB - The relation of the nerves with the piriformis muscle in the deep gluteal region examined in 50 buttock from 18 male and 7 male newborn cadavers. The nerves were found in usual position in 74% of sides, while one or more nerves perforated the piriformis in 16% and, unusual location of the nerves with intact piriformis was in 10% of sides. Abnormal located nerves are utilised in view of whether originating from dorsal or ventral part of the sacral plexus. PMID- 10693332 TI - [Effectiveness and adverse effects of sildenafil in erectile dysfunction]. AB - Sildenafil is a selective and by oral administration potent type-5 phosphodiesterase (PDE 5) inhibitor, which increases the erection by corpus cavernosum smooth muscle relaxation. In a non-placebo controlled study, 134 patients with erectile dysfunction were treated with oral sildenafil. The aim of the study was to estimate the efficacy and adverse effects of this treatment. 51 patients (38%) had psychogenic, and 83 (62%) organic origin of the erectile dysfunction. 73 of them have already had some treatment for this problem before. The effective dose was 50 mg for 84 patients (63%), 100 mg for 32 (24%) and 25 mg for 4 patients. The treatment was effective for 120 patients (90%). The most common adverse effect was flushing in 18 (13%) and headache in 9 (7%) cases, two patients had headache and flushing together. Nasal congestion and visual disturbances were complained by two patients. Two patients reported prolonged (max. 2h) erections. Cardiological investigation was performed for cardiovascular patients and for patients with risk factors. Exact criteria of the cardiological opinion of sildenafil treatment are reviewed. Cardial or other serious adverse effects were not observed. It was not necessary to stop the treatment because of the adverse effects. The authors found, that sildenafil is an effective and safe treatment for the erectile dysfunction. PMID- 10693333 TI - [Dynamic MR examination of the breast--histological correlations]. AB - The authors outline dynamic MR mammography (dMRM) as a highly sensitive diagnostic method for the examination of the breast. In a retrospective study relating to 84 processed cases, in the knowledge of the cytological-histological findings the diagnostic accuracy of the examinations was determined. The role of the method in detecting benign and malignant changes of the breast has been estimated. Misdiagnosed cases have been analysed and recommendations for the application of the method are included. The MR proved to be positive in 32 cases and negative in 3 cases of the analysed 35 malignant tumors. Benign lesions were found at microscopy in 49 cases, of which MR correctly diagnosed 40. The sensitivity and the specificity of dynamic MR mammography were 91% and 82%. PMID- 10693334 TI - [Neuroendoscopic removal of a colloid cyst of the third ventricle]. AB - The removal of the benign and only slightly symptomatic but by virtue of their localization potentially fatal lesions is a challenge for the neurosurgeons. Such cases demand a therapy without any postoperative morbidity. The neuroendoscopy meets these requirements and proves to be effective in more and more fields of the art. This article reports a case when colloid cyst was resected entirely by use of a neuroendoscope, firstly in Hungary. PMID- 10693335 TI - [Immunomodulation with fumaric acid. Systemic therapy in psoriasis]. AB - The treatment of psoriasis vulgaris with fumaric acid esters has been controversial for more than 30 years. Recently the fumaric acid derivatives are marketed antipsoriatics in many European countries. In this paper the clinical efficacy, the side effects as well as the mode of action of these highly potent substances are summarized. PMID- 10693336 TI - [A case of datura stramonium poisoning--general problems of differential diagnosis]. AB - In past year drug abuse becomes more and more general in Hungary. In addition to consume traditional drugs, other substances are used frequently too. One of them is the Datura stramonium, which contains alkaloids (mostly atropine), and can result in hallucinations. Therefore Datura stramonium is seemingly becoming popular as a hallucinogenic drug. The consumption of any part of the plant causes atropine intoxication, thus anticholinergic delirium. Differential diagnosis of Datura intoxication can be difficult in the everyday medical practise. In our paper the symptomatology, diagnosis, differential diagnosis, and therapy of Datura intoxication are discussed and we report one of our cases. PMID- 10693337 TI - [Changes in the parotid salivary gland of rats with experimental prostatitis]. AB - Experiments on 46 rats showed that infectious and allergic prostatitis led to development of dystrophic changes in the acinar tissue and ducts of the parotid glands. The cause of these changes is not clear; a humoral or neuroreflex relationship of a reactive type is suggested. PMID- 10693338 TI - [Hereditary pathology of the enamel and dentin. A review of molecular genetic research]. AB - Mapped phenotype of imperfect amelogenesis, type II imperfect dentinogenesis, hereditary opalescent dentin, Capdepont's dysplasia, and type II dentin dysplasia is described for the first time in Russia. Classification of hereditary disorders in dentin development is presented. PMID- 10693339 TI - [An enamel adhesion study of Esterfill adhesives with chemical and light hardening]. AB - The purpose of this work is to define the dependence of adhesive bond strength on a way of cure of adhesive system, application of the primer and bond in two step or in one step in vitro. Chemically curing "Esterfill bond" (AB) and both light curing primer and bond, and composite hybrid light-curing restorative material "Esterfill PHOTO" (EP) (DIAS LTD, Russia) were used. The shear bond strength was measured according to recommendation of ISO 11405:1994. The apparatus for these tests corresponds to ISO 10477, addition I. Shear bond strength was measured on enamel of 83 extracted human molars. Teeth were divided into 7 groups. Before using of adhesive system enamel was etched by 37% phosphoric acid during 60 sec. For measuring of shear bond strength the post composite restorative material "EP" (dentin) has been used. The received data show that the shear bond strength does nat depend on the type of cure, the composition of the bonds being similar. Using primer before application of the bond increases the shear bond strength approximately by 4-5 MPa. The inclusion of the primer in the content of the bond system increases the adhesion to enamel by 9 MPa. PMID- 10693340 TI - [The evaluation of periodontal status by the chemical composition of the oral media]. AB - Twelve patients aged 21-38 years with gingivitis and periodontitis and 9 subjects with intact periodontium were examined. Air from the oral cavity was collected with a special device, liquid samples were collected by gargling with sterile water. Chemical composition of the air and washings was analyzed by chromato-mass spectrometry, gas adsorption and gas liquid chromatography. Inflammation of periodontal tissues was associated with a sharp increase in the oral air concentration of methylethylketone. The concentrations of isovaleric, n-valeric, and n-enanthic aldehydes increased appreciably. Ethanol predominated over n-butyl alcohol. The levels of dimethylsulfide and isoprene increased several times. In the washings from the oral cavity, the content of microorganism's vital activity products (fatty acids) was increased. Possible metabolic mechanisms of the detected shifts are discussed. PMID- 10693341 TI - [The use of a cyanoacrylate-based composite material in operations on the alveolar processes of the jaws]. AB - Improvement of methods for preventing inflammatory complications after surgery on the jaw bones and induction of predictable regeneration of bone tissue at the site of intervention is an important problem. Numerous experimental and clinical studies in Russia and abroad demonstrated the efficacy of biodestroyed materials on the basis of rapidly polymerized monomers--cyanacrylates. The products of polycyanacrylate biodestruction are nontoxic and are not accumulated in the organism. Addition of antiseptics and antibiotics in sufficient amounts to polycyanacrylates prevents the development of inflammatory processes at the site of the composite application. A new Russian composite material MK-9M was used in 44 patients operated on the alveolar process bone at Department of Oral and Maxillofacial Surgery of Moscow Medical Stomatological Institute in 1997-1998. Pronounced anti-inflammatory and osteoregenerating effect was attained in all cases. PMID- 10693342 TI - [The presentation of relative values in scientific research results]. PMID- 10693343 TI - [The pathogenesis of posttraumatic deformities of the mandible in the growing organism]. AB - Experiments on developing animals showed that mandibular fractures of any location can be complicated by pronounced growth disorders and abnormal development of mandibular bone, leading to deformations of the site of fracture. Abnormal mandibular development was particularly obvious after injuries to the condyle and condylar process, after fractures of mandibular body and angle with dissection of the neurovascular bundle, and in cases with osteomyelitis. Disorders in endochondral bone formation (which is normally most active in the condyle) caused by traumatic processes and disorders in neurotrophic regulation and blood supply to the bone, developing in neurovascular obstruction, underlie the mechanisms of posttraumatic mandibular deformations. These complications were observed in patients with fractures of different sites because of improper joining and fixation of fragments. Children should be regularly checked up after mandibular fractures. Measures aimed at prevention of growth disorders and development of mandibular bone lesions are obligatory at all stages of examination. PMID- 10693344 TI - [The clinical x-ray aspects of the diagnosis and treatment of the temporomandibular joint pain dysfunction syndrome]. AB - Clinical and x-ray picture of the syndrome of painful dysfunction of the temporomandibular joint is described. Differential diagnosis of this syndrome and other articular diseases with similar clinical symptoms is presented. Treatment strategy is described in brief. PMID- 10693345 TI - [The clinico-microbiological evaluation of the efficacy of using new drug forms of chlorhexidine--Corsodyl and Eludril--for the prevention of infectious complications in operations for endosseous implantation]. AB - Prevention of infectious inflammatory complications of intraosseous implantation is a pressing problem. Chlorohexidine dosage forms Corsodyl and Eludril were used for this purpose. In the control group, furacillin was used. Microbiological studies showed that instillations of Corsodyl and Eludril solutions during the postoperative period did not modify normal oral microflora and led to disappearance of the most aggressive anaerobic bacteria in 1-3 days. These drugs are more effective that Listerine, and they are recommended for prevention of infectious inflammatory complications after intraosseous implantation. PMID- 10693346 TI - [The clinical picture, diagnosis and radiation treatment principles in cancer of the buccal mucosa]. AB - Diagnosis and treatment of 228 patients with cancer of the buccal mucosa is analyzed. There were 130 women and 98 men aged 65 years on average. 218 were primary patients and 10 with relapses. Squamous-cell carcinomas with hornification predominated: 217 cases (95%). Adenocarcinoma was diagnosed in 8 (3.5%) and poorly differentiated cancer in 3 patients. Metastases to the regional lymph nodes were detected in 25% cases. For cosmetic reasons, radiotherapy (oral x-ray therapy, interstitial method, long-distance gamma beam therapy) was the method of choice. PMID- 10693347 TI - [The use of computer technologies in rehabilitative and reconstructive surgery on the facial supporting tissues]. PMID- 10693348 TI - [The x-ray stereometric assessment of mandibular asymmetries]. AB - The method of spatial appreciation of mandibular structure on the basis of straight and lateral cephalogram is proposed. Asymmetry is appraised by comparison of its right and left side. The analysis is based on linear and corner sizes. Possibilities and perspectives of present line of research are outlined. PMID- 10693349 TI - [Endoscopic technologies in maxillofacial surgery]. AB - Endoscopic operations in 107 patients with numerous maxillofacial diseases are analyzed. Endoscopic technologies were used in diseases of the maxillary sinus, temporomandibular joint, injuries to the facial nerve, and cysts of the jaws. New methods for surgical treatment of mandibular fractures, zygomatico-orbital complex, cysts of the jaws are proposed. The results of utilization of endoscopic technologies in maxillofacial surgery are analyzed and their high efficacy and good prospects are demonstrated. PMID- 10693350 TI - [Impaired occlusion is the main etiological factor in the occurrence of temporomandibular joint dysfunction]. AB - A homogeneous group of 104 subjects aged 21-34 years was examined. Forty-nine (47%) subjects (study group) presented with symptoms of abnormalities of the masticatory muscles and temporomandibular joint, 58 subjects (53%) without such symptoms were controls. Clinical examination was carried out with evaluation of occlusion, Angle's [correction of Engle's] class, and analysis of dental contacts in the oral cavity in different types of occlusion. Disorders of occlusion were detected in 31% of controls and in 73% patients in the study group. Disorders of occlusion correlated with the number of symptoms of temporomandibular joint dysfunction. The detected super-contacts of teeth are the main etiological factors leading first to discoordination of the masticatory muscles and then to functional disorders of the temporomandibular joint. PMID- 10693351 TI - [The dynamics of the colonization by the microbial flora of the mouth of the different materials used for dental prosthetics]. AB - Colonization of materials used for making dentures with oral microflora was studied in 12 patients with chronic generalized medium severe periodontitis during remission. Time course of dentures colonization with representatives of oral microflora (S. sanguis, Peptostreptococcus sp., Prevotella oralis) and with bacteria pathogenic for the periodontium (Prevotella melaninogenica, A. naeslundii, Fusobacterium sp.) depends on the material from which the dentures are made. Minimal colonization of bacteria pathogenic for the periodontium was observed on cermet dentures. PMID- 10693352 TI - [The classification of congenital clefts and defects of the palate after uranoplasty. The clinico-embryo-pathogenetic principles]. AB - The authors suggest classifying palatal clefts as an individual nosological entity resultant from impaired differentiation, growth, and fusion of primary embryonal structures forming the palate as an anatomical structure. Definition of the palatal interface on the basis of the embryo-pathogenetic principle underlies the working classification of palatal defects developing after uranoplasty. PMID- 10693353 TI - [Reconstruction of the hard palate in congenital clefts]. AB - Cleft palate advancement is linked with the new bone regeneration after palate and alveolar process osteotomy and underdeveloped palatal process of the jaw moving to a normal anatomy position. PMID- 10693354 TI - [The expert evaluation of dental institutions during their licensing and accreditation]. PMID- 10693355 TI - [The evolution of the concepts on the causes of the occurrence of dental caries]. PMID- 10693356 TI - [The 50th anniversary of the awarding of the State Prize in the Field of Medicine to A. A. Limberg]. PMID- 10693357 TI - [Prof. B. M. Pashkov--the founder of the dermatostomatological direction in the solution of the scientific problems of dentistry (on the centenary of his birth)]. PMID- 10693358 TI - Hepatitis A vaccines. PMID- 10693359 TI - Who pays the bill and who makes the profit in treating chronic disease? PMID- 10693360 TI - Who should care for people with chronic diseases? PMID- 10693361 TI - The epidemic of obesity. PMID- 10693362 TI - Compensation awarded for death after illegible prescription. PMID- 10693363 TI - HMO sues charity over domain name. PMID- 10693364 TI - Ask patients which herbal drugs they smoke as well as eat. PMID- 10693365 TI - Clinicians in private practice give generously in teaching time. PMID- 10693366 TI - A cutaneous manifestation of a systemic disease. PMID- 10693367 TI - Case-control study of 10 years of comprehensive diabetes care. AB - OBJECTIVE: To describe the long-term clinical impact of a comprehensive management program instituted throughout a health system for members with diabetes mellitus. DESIGN: 10 year case-control evaluation. SETTING: Kaiser Permanente Northwest, Portland, OR. PARTICIPANTS: Members of the health maintenance organization between 1987 and 1996; members with diabetes were compared with equal numbers of members without diabetes. The number of participants with diabetes ranged from 5331 in 1987 to 13,099 in 1996. MAIN OUTCOME MEASURES: Number in diabetes register, mortality, change in comorbidity, rates of uptake of preventive health measures, use of pharmaceuticals, levels of risk factors, hospital days per thousand per year, emergency room visits per thousand per year. RESULTS: The prevalence of diabetes identified in this population rose from 2.54% (7,895/310,819) in 1987 to 3.66% (14,741/402,754) in 1996, and the mean (SEM) age of members at the time of diagnosis fell slightly from 62.9 (+/- 0.21) years to 62.0 (+/- 0.13) years (P < 0.05). By 1996, 10,885 of the 13,099 (83% +/- 0.3%) of members with diabetes had an annual laboratory test to assess glycemic control, the annual screening rate for retinopathy was 67.6% (+/- 0.4%), the rate of uptake of influenza immunizations was 60.2% (7,886/13,099) and the screening rate for nephropathy was 43% (5,698/13,099) (+/- 0.49%). The use of home glucose testing increased from 32.4% (1721/5331) of members with diabetes to 53.0% (6,942/12,099); the use of lipid lowering drugs increased from 3.5% (187/55,331) to 19.8% (2,594/13,099). The use of angiotensin converting enzyme inhibitors increased from 8.5% to 34.8% of members with diabetes. Mean blood pressure decreased from 144/82 mm Hg (+/- 0.8/0.4) to 138/79 mm Hg (+/- 0.3/0.15), and mean total cholesterol concentrations dropped from 243 mg/dL (+/- 4.2) to 215 mg/dL (+/- 0.6). By 1996, 56.4% (7,388/1,3099) (+/- 0.5%) of members on the diabetes register had good to excellent glycemic control (HbA1c < 8%). Mortality decreased from 4.8% (256/5331) (+/- 0.3%) to 3.6% (472/13,099) (+/- 0.2%) among members with diabetes, this was a more rapid decrease than was observed among those without diabetes (P < 0.01). The annual ratio of visits to the emergency room by members with diabetes to members without fell from 2.5 to 1.8, and the ratio for the number of days spent in acute care in the hospital dropped from 3.6 to 2.5. CONCLUSIONS: This centrally organized program based in a primary care setting and utilizing a register of patients with diabetes was associated with substantial improvements in the process and outcomes of care in a large population of health maintenance organization members with diabetes. PMID- 10693368 TI - Randomized, controlled trial of glucosamine for treating osteoarthritis of the knee. AB - OBJECTIVE: To determine the effectiveness of glucosamine in reducing pain from osteoarthritis of the knee. DESIGN: Randomized, double-blind parallel trial of glucosamine 500 mg three times daily or a placebo for 2 months. SETTING: Veterans Affairs Medical Center, Prescott, AZ. PARTICIPANTS: Ninety-eight patients aged 34 to 81 being treated for osteoarthritis of the knee. MAIN OUTCOME MEASURES: Pain intensity both at rest and while walking as assessed by a visual analog scale at baseline and after 30 and 60 days of treatment. RESULTS: Forty-nine patients were randomly allocated to each group. There was no statistical difference between the two groups in scores on the visual analog scale at 30 days for resting (mean [SD] score placebo group 3.5 [2.7] vs 3.3 [2.4] glucosamine group, P = 0.66) or walking (5.1 [2.6] vs 5.3 [2.4], P = 0.69); there was also no difference at 60 days for resting (3.4 [2.5] vs 3.2 [2.5], P = 0.81) or walking (4.9 [2.2] vs 4.9 [2.8], P = 0.90). There was also no statistical difference between groups in the mean change from baseline in scores on the visual analog scale (mean [SD] change for walking at 60 days placebo group -1.5 [2.5] vs glucosamine group -1.4 [3.0], P = 0.77). Two participants taking glucosamine and 4 taking placebo withdrew from the study due to adverse side effects (P = 0.67). CONCLUSION: Glucosamine was no better than placebo in reducing pain from osteoarthritis of the knee in this group of patients. PMID- 10693369 TI - Use of alternative products: where's the beef? PMID- 10693370 TI - Magnesium sulfate is effective for severe acute asthma treated in the emergency department. PMID- 10693371 TI - Ovarian cancer screening was feasible but did not decrease incidence of index cancer or mortality. PMID- 10693372 TI - How medications affect thyroid function. PMID- 10693373 TI - Use of opioids to treat chronic, noncancer pain. PMID- 10693374 TI - Update on Alzheimer's disease: recent findings and treatments. PMID- 10693375 TI - Chronic back pain: does bed rest help? PMID- 10693376 TI - Emotional dimensions of chronic disease. PMID- 10693377 TI - How to deal with medically unknown symptoms. PMID- 10693378 TI - When AIDS became a chronic disease. PMID- 10693379 TI - Individualized stepped care of chronic illness. PMID- 10693380 TI - Speaker's corner. Living with thalassemia. PMID- 10693381 TI - In favour of tolerance. PMID- 10693382 TI - Hyperventilation and the body. AB - Hyperventilation has rapid and far-ranging physiological effects via its alteration of pH and depletion of CO2 in the body, resulting in respiratory alkalosis, acute or chronic. The general effects on skeletal and smooth muscles, as well as neural tissue, are summarized. A wide variety of symptoms such as pain, tension, disturbances of consciousness, circulatory problems and cardiovascular effects can confound treatment efforts but may respond to alterations of breathing patterns. Suggestions are given for detecting this condition. PMID- 10693383 TI - It's a disaster: emergency departments' preparation for a chemical incident or disaster. AB - Nurses in the Accident & Emergency (A&E) Department have a significant role to play in the treatment and resuscitation of victims of a chemical disaster. Chemical disasters are unique because casualties are contaminated. Nursing staff triage casualties and they have direct contact with contaminated patients, before and during decontamination. Consequently they require adequate personal protective equipment and information regarding isolation and decontamination. The use of chemicals has increased since the turn of the century. Hazardous chemical emergencies arise from accidents in production, storage, transportation and the disposal of chemical substances. Their illegal manufacture and use by terrorists makes the likelihood of a chemical disaster with mass casualties in Australia very real. Emergency departments are ill-prepared to deal with this scenario, and very few disaster plans include a comprehensive decontamination component. To achieve an effective response with the best utilisation of resources, it is vital for emergency services personnel and A&E departments to be prepared. PMID- 10693384 TI - Fundamental issues in questionnaire design. AB - The questionnaire is probably the most common form of data collection tool used in nursing research. There is a misconception that anyone with a clear grasp of English and a modicum of common sense can design an effective questionnaire. Contrary to such common belief, this article will demonstrate that questionnaire design is a complex and time consuming process, but a necessary labour to ensure valid and reliable data is collected. In addition, meticulous construction is more likely to yield data that can be utilized in the pursuit of objective, quantitative and generalizable truths, upon which practice and policy decisions can be formulated. This article examines a myriad of fundamental issues surrounding questionnaire design, which encompass question wording, question order, presentation, administration and data collection, amongst other issues. PMID- 10693385 TI - Responding to autonomy and disempowerment at the time of a sudden death. AB - This paper examines the difficulty of giving people choices and recognising their autonomy at the time of a sudden death. It discusses how they achieve this whilst disempowered and frightened. PMID- 10693386 TI - Increasing patient numbers: the implications for New Zealand emergency departments. AB - Within the New Zealand (NZ) health care system, a number of changes have affected the way health care is accessed and delivered. Emergency Departments (EDs) are noticing increased attendance of patients with minor or non-urgent conditions. This increase in patient volume, together with on-going fiscal constraints and restructuring, has placed an added strain on the functioning of EDs. New Zealand nurses need to question the role currently given to EDs and identify the issues surrounding the increased use of these departments for primary health care. Is this move feasible in the NZ environment, and what are the implications for emergency nurses? PMID- 10693387 TI - Accident prevention in the A&E department. AB - Accident and Emergency units are often regarded as the 'shop window' of a health service. While the priority is to attend to those in need of emergency services, there may also be opportunities for health education amongst a 'captive audience', particularly in relation to accident prevention. This report illustrates how the influence of one A&E department was used amongst primary school children to increase their awareness of cycle safety. Educational materials were developed, a series of visits to schools was planned, which over 600 children attended, and a competition organised. Initial responses from the children were very positive, and an analysis of subsequent cycle injury levels is now under way. The project formed part of a wide ranging practice development programme within a large district general hospital in Whitehaven in collaboration with St Martin's College Department of Nursing. It combined the need to develop quality services and staff in tandem. The outreach educational approach is now being examined for adoption in other areas of the Trust. PMID- 10693388 TI - Reflective narrative: Autonomous Practitioner course. PMID- 10693389 TI - Farewell to A&E. AB - Carolyn Stead's moving account of the sudden death of her baby son Dominic was published in the January 1998 issue. It is reprinted on pages 170-174, before this update, in which Carolyn outlines her subsequent experiences and emotions. PMID- 10693390 TI - A classical dilemma. PMID- 10693391 TI - Valuing nurse practitioners. PMID- 10693392 TI - The faculty of emergency nursing. A personal view. PMID- 10693393 TI - The 'millennium bug'. Its effect on the workplace and what is being done about it. PMID- 10693394 TI - Antifreeze poisoning. PMID- 10693395 TI - Inhalation injuries. PMID- 10693396 TI - Autonomous practitioner or handmaiden? PMID- 10693397 TI - Training users of telemedicine. PMID- 10693398 TI - Managing managed care: the next level for transcultural nurses. PMID- 10693399 TI - Transcultural nursing and community-based care. PMID- 10693400 TI - Voices of transcultural nursing. PMID- 10693401 TI - From a practice perspective. PMID- 10693402 TI - Response to the commentaries on defining transcultural nursing. PMID- 10693403 TI - Nursing and anthropology: two worlds to blend. PMID- 10693404 TI - Global perspectives in nursing and health. PMID- 10693405 TI - Health promotion in cervical cancer prevention among the Yakama Indian women of the Wa'Shat Longhouse. AB - The purpose of this 3-year study was to gain a greater understanding of the importance of the Wa'Shat Longhouse religion to the design of a culturally appropriate health promotion (cervical cancer prevention) program with the Yakama Indian people of eastern Washington. This descriptive study involved interviews with 10 Wa'Shat members, observations, and participant observations of 30 community ceremonial activities. The framework of health promotion planning guided the investigation. We found that (a) program goals needed to be holistic and wellness oriented, (b) teaching methods needed to include circular symbols, and (c) intervention strategies needed to be linked to the natural patterns of communication of the Wa'Shat Longhouse and to involve elders. Storytelling, talking circles, and use of role models were all found to be important teaching methods. We confirm previous perspectives on the importance of religion, provide greater depth in this understanding and outline implications for transcultural nursing practice. PMID- 10693406 TI - Transcultural nursing with Native Americans: critical knowledge, skills, and attitudes. AB - Native American nurses are in an ideal position to articulate how to serve Native people in ways that respect indigenous cultures. Forty Native American nurses and nursing students completed a survey on knowledge, skills, and attitudes/values necessary for culturally competent service provision to Native patients. Four knowledge themes, four skill themes, and three attitude/value themes were identified. Nurses must know about and show respect for the culture and history of Native groups, including culturally specific health beliefs and healing practices. Skills must be tailored for work in a non-Western context, and containment skills become particularly important. Additionally, respecting diversity and traditions and being open-minded and nonethnocentric are fundamental. This study provides valuable information nurses can use to enhance their work with indigenous people. PMID- 10693407 TI - Mexican American women's expressions of the meaning of culturally congruent prenatal care. AB - The purpose of this study was to describe, explicate, and systematically analyze the meanings, expressions, and experiences of generic and professional care during pregnancy of Mexican American women in their home and prenatal clinic contexts. The ethnonursing method was used to describe cultural care among 16 key and 34 general informants. Generic culture care was valued and considered essential by informants for healthy childbearing. Six major themes were synthesized from 30 cultural patterns: (a) generic culture care included the protection of the mother and fetus by older Mexican American women and was greatly influenced by religion and family beliefs and practices; (b) generic culture care was family obligation for provision of filial (family) succorance, sharing of self, and being with the childbearing mother; (c) culture care was respect for familial caring roles in relation to age and gender; (d) childbearing Mexican American women viewed culture care by professional nurses as concern for, professional knowledge, protection, being attentive to, and explaining; (e) culture care was use of the Spanish language in caring interactions in diverse environmental contexts; and (f) professional prenatal care was valued by Mexican American women and was influenced by legal, economic, and technological factors in the social structure. PMID- 10693408 TI - Intergenerational analysis of dietary practices and health perceptions of Hispanic women and their adult daughters. AB - This descriptive, correlational, two-group study investigated differences between dietary practices, acculturation, and health perceptions in a convenience sample of Hispanic mothers and their adult daughters (N = 76, 47 mother-daughter dyads). Analysis (paired t tests) of the Block Screening Questionnaire, General Acculturation Index, and Self-Rated Health Subindex of the Multilevel Assessment Instrument showed significant differences: Daughters ate more fat (p = .04) and were more acculturated than their mothers (p = .0001). The Pearson correlation yielded a significant relationship for the 76 subjects between fat intake (dietary practice) and health perception: The more fat (meat/snacks) intake, the more negatively women perceived their health status (p = .0001). PMID- 10693409 TI - A grounded theory study of the experience of type 2 diabetes mellitus in First Nations adults in Canada. AB - Diabetes among First Nations peoples has reached epidemic proportions, and diabetes prevalence, complications, and mortality rates are higher than in Whites. The purpose of this grounded theory study was to investigate the experience of Type 2 diabetes in First Nations adults. Ten individuals living in one reserve community, in southwestern Ontario, participated in this research. Participants were interviewed on two occasions by a First Nations nurse. All interviews were audiotaped and transcribed verbatim. A three-phase process of integration emerged, similar to the process of integration discovered in prior research with Type 1 diabetic Whites. However, there were some important differences in the characteristics of these three phases. In addition, there were surprising findings related to beliefs about the type of diabetes educator desired by these First Nations clients. PMID- 10693410 TI - Beliefs about childhood immunisation among Lebanese Muslim immigrants in Australia. AB - The aim of the study was to describe and analyse care values, beliefs, and practices relating to immunisation by Lebanese Muslim immigrants in New South Wales (NSW), Australia. This ethnonursing study explored the importance of care related to immunisation, knowledge of informants relating to vaccines, diseases, side effects, and contraindications. Family responsibilities relating to immunisation care services as well as expectations and evaluations of care services provided were also examined. Data were collected via observation participation-reflection, including in-depth interviews. The findings revealed significant care themes for Lebanese Muslim informants based on their cultural values, beliefs, and practices related to health and immunisation. Culturally congruent nursing care practices related to immunisation for Lebanese Muslims in NSW, Australia, were identified. PMID- 10693411 TI - From nurse to nurse anesthetist: the relationship of culture, race, and ethnicity to professional socialization and career commitment of advanced practice nurses. AB - The purpose of this study was to identify the relationship of culture, race, and ethnicity on professional socialization and career commitment of student registered nurse anesthetists. A 78-item, self-administered questionnaire was mailed to the United States population of student nurse anesthetists (N = 2,008) yielding a 56% response. Demographic data and the dimensions of socialization were analyzed with chi-square and weighted least-squares linear analysis of variance. Four scales assessed the relationship of age, gender, culture/race/ethnicity, and views of culturally congruent care on the socialization process. In all dimensions of socialization, culture and race/ethnicity both correlated significantly (p < .05) for all nondominant groups compared to the dominant European American White group. Responses from Asian Pacific Islanders were most positively correlated to all dimensions of socialization. Hispanics responded least positively to a life-time commitment to the career in nurse anesthesia by culture (p = .003) and ethnicity (p = .009). PMID- 10693412 TI - Interpreters: a double-edged sword in nursing practice. AB - The provision of health services for all Australians is based on equality of access to health care services, regardless of cultural origin or linguistic skill, and on the responsibility of the health system to respond appropriately. Lack of fluency in the English language and lack of bilingual or multilingual nurses are major sources of miscommunication, even with the use of interpreters and translated health information. Many immigrant women find the use of interpreters unacceptable. Nurses are concerned with legal issues. These two viewpoints make the use of interpreters a double-edged sword in practice. The findings of recent research and the literature in relation to language skills and the use of interpreters and translations in the Australian context are explored. Potential resolutions for transcultural nursing practice are provided. PMID- 10693413 TI - Founder's focus--faculty limit students' study of transcultural nursing: a critical issue. PMID- 10693414 TI - Pain as the fifth vital sign: will cultural variations be considered? PMID- 10693415 TI - Transcultural nursing's role in a managed care environment. PMID- 10693416 TI - Culturally competent managed health care: a family physician's perspective. PMID- 10693417 TI - A framework for providing culturally competent health care services in managed care organizations. PMID- 10693418 TI - Managed care: a turning point for nursing. PMID- 10693419 TI - Let us try to keep culturally competent care in managed care. PMID- 10693420 TI - Managed care: pitfalls for cultural bias. PMID- 10693421 TI - The personal experience of pregnancy for African-American women. AB - This study describes the personal experiences of pregnancy for African-American women. Data were obtained from two group interviews with four African-American nurse-midwives who had experienced pregnancy and had extensive professional experience in the provision of health care services to pregnant African Americans. Three major themes were constructed from the interview narratives. The first concerned the experience of pregnancy as a transition experience from childhood to adulthood and from womanhood to motherhood, involving heightened senses of maturity, self-esteem, and intimacy. The second identified stresses experienced by African-American women, including the lack of material resources and emotional support. The last theme concerned the provision of effective support in pregnancy. The significance of interpersonal relationships with the pregnant women's mothers, other significant women, and their partners was described. Implications for practice included suggestions for the provision of effective emotional support from health care professionals such as attentive listening and the elimination of environmental factors that communicate lowered personal value. PMID- 10693422 TI - Developing an instrument to study the tuberculosis screening behaviors of Mexican migrant farmworkers. AB - This article details the use of qualitative data in the construction of a Spanish language, quantitative research instrument designed for a study of Mexican migrant farmworkers' participation in tuberculosis screening. In the qualitative study, 19 Mexican migrant farmworkers were interviewed in Spanish to elicit their explanatory models (EMs) about tuberculosis. The Tuberculosis Interview Instrument (TII) was developed from the results of the qualitative study and concepts from a theoretical framework consisting of a combination of the Health Belief Model (HBM) and the Theory of Reasoned Action (TRA). After its development, the TII was subjected to translation and back-translation procedures to insure the equivalency of the English and Spanish versions, and it was reviewed for content validity. Internal consistency reliability, based on the responses of 206 subjects, was satisfactory for all subscales. Future testing of the TII is recommended to evaluate its stability among Mexican migrant farmworkers in other parts of the United States. PMID- 10693423 TI - The health care meanings, values, and practices of Anglo-American males in the rural midwest. AB - Limited nursing knowledge exists to explain how culture influences men's health care decisions and practices. The purpose of this collaborative research was to explore the health care meanings, values, expressions, lived experiences, and practices of Anglo-American adult males residing in the rural Midwest. Using the ethnonursing research method, data were collected through interviews with 7 key and 12 general informants residing in Iowa. Four culture values and five themes were supported by the findings. Leininger's culture care modes were used to explicate culturally meaningful nursing care actions and decisions that are in harmony with the cultural values and practices of rural men. PMID- 10693424 TI - Panamanians' practices for health promotion and the meaning of respect afforded them by health care providers. AB - This study describes and compares Panamanians' and Panamanian Americans' practices for health promotion and wellness, disease and illness prevention, and the meaning of respect afforded them by health care providers. Understanding a person's beliefs and values when planning nursing and health care interventions helps the caregiver provide culturally acceptable care that improves clients' satisfaction and health status. Culturally respectful, acceptable, and appropriate care conserves the utilization of human, material, and financial resources. This sample comprised 70 subjects: 50 in the Republic of Panama and 20 from the DelMarVa Peninsula in the United States. This study uses selected primary and secondary characteristics of culture and selected domains from the Purnell Model for Cultural Competence as guides for questionnaire development, data analysis, and discussion of the findings. PMID- 10693425 TI - Disseminating the results of participant-focused research. AB - Participant-focused research (PFR) includes the "subjects" as full partners in the research process. As such, participants share in the products or outcomes of research. PFR goes beyond the traditional research approach of disseminating findings to other scientists and clinicians and includes participants and community residents in sharing the skills, knowledge, and resources of the study with the objective of empowering the participants. This article demonstrates the use of PFR in disseminating the results or products of study to the participants through two examples of long-term research projects conducted in Los Angeles. The first example is a community-based study of HIV prevention with low-income Latina women. The second example is an ethnographic study of health concerns and risks among adolescents in juvenile detention. These examples provide two approaches to dissemination of research findings and benefits to the participants and the community. PMID- 10693426 TI - The culture of the deaf. AB - The population of the United States includes over 1 million deaf people, the majority of whom have chosen to differentiate themselves in terms of a Deaf culture. These Deaf people share unique values and norms and use sign language for communication. The differences that exist between hearing and Deaf individuals necessitate that the nurse understand this population. This article will describe the Deaf culture using Leininger's Sunrise Model and present specific nursing actions in the modes of culture care preservation and/or maintenance, accommodation and/or negotiation, and repatterning or restructuring that enable the nurse to provide culturally congruent care for Deaf clients. PMID- 10693427 TI - People with disabilities--the same, but different: implications for health care practice. AB - Despite the author's experiences as a nurse and parent of a young adult with physical disabilities, the author had much to learn about the culture surrounding disability. The contemporary "minority-group" model for disability replaces a medical model that views people with disabilities in need of remediation. Disability rights advocates often criticize health professionals, citing erroneous assumptions and failure to understand the perspectives of disabled persons. The author illustrates applications for clinical practice by health professionals with excerpts from a qualitative study that explored the spiritual experiences of adults with physical disabilities and family members as well as their responses to lived experience with disability. PMID- 10693428 TI - [Living worlds of children with special need for support between institutions and daily integrated assistance]. PMID- 10693429 TI - [Screening of newborns--meaningful management]. PMID- 10693430 TI - [Engagement of an outsider at the therapeutic education center Hochbend in Tonisvorst--report of a student's practical training in the pediatric nursing school at the Krefeld Hospital]. PMID- 10693431 TI - [Gentle care of prematures--their care in the premature unit]. PMID- 10693432 TI - [Evaluation of pediatric nursing education based on a modified "Oelke curriculum"]. PMID- 10693433 TI - [Engagement instead of heroism. Delegates to the German Federation of Pediatric Nurses--persons, scope of activities, range. Project--Part 2: Structure of delegation]. PMID- 10693435 TI - [Letters on the topic: Babies, lopsided and crookes in the carrying cloth, in "Kinderkrankenschwester", from 8/99, page 336] PMID- 10693434 TI - [Professor Hellbrugge on his 80. birthday (10/23)]. PMID- 10693436 TI - [How long to use antibiotics therapy in acute otitis media?]. PMID- 10693438 TI - [The ideal mother]. PMID- 10693439 TI - [Understanding, phenomenologic-biographical diagnosis--an alternative to traditional classification and diagnostic systems in nursing?]. PMID- 10693437 TI - [Viruses in otitis media]. PMID- 10693440 TI - [Care after a drowning accident]. PMID- 10693441 TI - [The ideal children's hospital. Children's Hospital-Image and Benchmarking study Germany ]. PMID- 10693442 TI - [20 years of ambulatory pediatric care in Germany--development and current situation]. PMID- 10693443 TI - [Self-induced illnesses: the Munchhausen syndrome]. PMID- 10693444 TI - [The counter-reformation]. PMID- 10693445 TI - [The pediatric hospital of the Erlangen-Nurnberg University presents its nursing targets]. PMID- 10693446 TI - [Why healthy sleep is so important. Report from the 7. German Congress for Sleep Research and Sleep Medicine in Dresden]. PMID- 10693447 TI - [Well taken care of]. PMID- 10693448 TI - [Perioperative therapy problems. Alcoholics in operative medicine--with special regard to the preoperative alcohol withdrawal syndrome]. PMID- 10693449 TI - [Patients as customers]. PMID- 10693450 TI - [Highest quality, yet lower costs]. PMID- 10693451 TI - ["A dive into cold water". Civil servants in nursing and caring]. PMID- 10693452 TI - [Alzheimer's disease]. PMID- 10693454 TI - [Kidney failure and therapeutic possibilities]. PMID- 10693453 TI - [Baxter Healthcare is starting HomeChoice PRO]. PMID- 10693455 TI - [The first. enteral nutrition, especially designed for the elderly is presented at the Reha in Dusseldorf]. PMID- 10693456 TI - [Glass, the stuff from which bottles are made]. PMID- 10693457 TI - [OptiPen Pro--more safety for Insulin injections]. PMID- 10693458 TI - [Development of a profession. The hypertension nurse]. PMID- 10693459 TI - [House-dust allergy: many recommendations are helping industry, not the patients]. PMID- 10693460 TI - [Shaken by fate--but I won't give in]. PMID- 10693461 TI - [Growing impotence] PMID- 10693462 TI - [Founding of the research group "Monastery Medicine". Mediator between historical healing and modern medicine]. PMID- 10693464 TI - [Legal aspects of palliative care]. PMID- 10693463 TI - [Palliative medicine. The Dutch euthanasia model has failed]. PMID- 10693465 TI - [Special features in diagnosis and therapy of pulmonary diseases in the aged]. PMID- 10693466 TI - [Strict indications for transurethral indwelling bladder catheters in general surgery]. PMID- 10693467 TI - [Hepatitis C more dangerous than AIDS. At last a breakthrough in therapy using the new combination treatment]. PMID- 10693468 TI - [Viruses]. PMID- 10693469 TI - [New: Biatain foam dressings--for wounds with copious exudate]. PMID- 10693470 TI - [Hodgkin's disease: light in the darkness]. PMID- 10693471 TI - [Preservation of life--from inclination or obligation?] [In Process Citation] PMID- 10693472 TI - [When do I need psychotherapy?]. PMID- 10693473 TI - Notes from England. PMID- 10693474 TI - Beyond Pierre Robin sequence. AB - The label Pierre Robin sequence is given to infants presenting with a triad of specific congenital anomalies: micrognathia, glossoptosis, and cleft palate. However, this label should be considered the first, not the final, step in the diagnostic process. In approximately 80 percent of newborns with Pierre Robin sequence, the triad of anomalies is part of an underlying genetic condition. This article reviews the variable etiologies of and general clinical considerations for Pierre Robin sequence. To illustrate how clinical management might vary based on the identification of an underlying condition, three case examples of neonates with Pierre Robin sequence and different underlying genetic conditions are presented. PMID- 10693475 TI - Adrenal insufficiency in the term and preterm neonate. AB - The adrenal gland is part of the endocrine system and produces hormones that are essential for survival. The adrenal cortex, the largest part of the adrenal gland, produces three major hormones. Glucocorticoids regulate metabolism of glucose, protein, and fat and allow the body to respond to stress by increasing blood glucose levels and cardiac output. Mineralocorticoids regulate fluid and electrolyte balance and help maintain blood pressure. Androgens are responsible for sexual differentiation in the fetus. Insufficient production of these hormones can occur for many reasons and can have a profound effect on the newborn. It is important for neonatal nurses to be familiar with signs and symptoms and appropriate treatment of adrenal insufficiency. This article explores causes, signs and symptoms, and treatment of adrenal insufficiency in the newborn. PMID- 10693476 TI - Back to sleep: is there room in that crib for both AAP recommendations and developmentally supportive care? AB - This article offers one institution's approach to implementation of the recommendations for infant sleep positioning as set forth by the American Academy of Pediatrics. The guidelines are directed toward healthy infants in the first year of life, a population not always encountered by the neonatal nurse. The guidelines focus on supine sleep position and the minimization of additional bedding, both of which can be challenging when contrasted with accomplishing supportive positioning and the goals of developmentally supportive care for ill or preterm infants. A multidisciplinary task force was formed to consider this challenge. The outcome is an evidence-based policy that is presented as an example for other clinicians. The policy addresses the following major components: sleep position with specific clinical exceptions, the use of bedding materials, play position during awake states, and parent education with preparation for discharge. The article also outlines the process by which the task force plans to implement and evaluate necessary practice changes. PMID- 10693477 TI - Suspended mothering: women's experiences mothering an infant with a genetic anomaly identified at birth. AB - PURPOSE: To develop an understanding of the experiences of women who give birth to an infant with a genetic anomaly not identified during the prenatal period. DESIGN: Exploratory design using selected qualitative methods. SAMPLE: Seven mothers of liveborn infants with a genetic anomaly characterized by physical attributes evident at birth. MAIN OUTCOME VARIABLE: Mother's expressions of suspended mothering. RESULTS: Two themes emerged that characterize the experience of suspended mothering: diminished maternal role and false protection. PRACTICE RECOMMENDATIONS: Facilitating maternal involvement in decision making requires giving women unbiased information in a supportive manner. PMID- 10693478 TI - Feeding with an umbilical arterial line. PMID- 10693479 TI - Peripheral pulmonary stenosis murmurs. PMID- 10693480 TI - A case study of antenatal distress and consequent neonatal respiratory distress. PMID- 10693481 TI - Shared learning: a phenomena for celebration and a challenge to health educators. PMID- 10693482 TI - The use of brief written educational material to promote reflection amongst trained nurses: a pilot study. AB - The UKCC (1995) suggest that in order for nurses to maintain their registration, they must maintain a personal professional profile via reflection, and there are numerous approaches to the teaching of reflection. Similarly, several models of reflection exist. However, much of this activity has occurred without attempts to establish the effectiveness of either reflection or attempts to teach it. The current study attempted to examine whether written educational material could significantly improve nurses' reflective ability. Nineteen practising nurses were recruited from a variety of sources and offered an educational package, largely based on Boud et al. (1985) model of reflection in learning. In a repeated measures design study, these participants completed a reflective exercise before and after offering of the educational material. Their scripts were rated, using Wong et al.'s (1995) tool, by three raters, two of whom were unaware of whether the scripts were completed before or after offering of the educational material. An initial analysis showed that there were no statistically significant changes in subjects' reflective ability following education. However, removal from the analysis of 6 participants who had a pre-test ability at the highest possible level (and for whom no improvement was, therefore, possible) resulted in a significant improvement in participants' ability from pre- to post-test (z = 2.4450, P = 0.0145). Interrater reliability calculated using Cronbach's Alpha was 0.7210 for pretest reflective accounts and 0.7369 for post-test reflective accounts. PMID- 10693483 TI - Designing and implementing a 'dual diagnosis' module: a review of the literature and some preliminary findings. AB - People with a learning disability experience the full range of mental health problems, with prevalence greater than in the general population. The literature suggests that the move to community care has highlighted their special needs, as they face a complexity of life experiences and risks. Nurses play a key role, and must be equipped with appropriate knowledge and expertise to respond to the mental health problems of people with a learning disability ('dual diagnosis'). Major national initiatives support research and propose access to appropriate education for care professionals. In order to respond to locally identified educational needs a 'dual diagnosis' module was developed for nurses working with people with a learning disability who experienced mental health difficulties. The aims were to increase knowledge, to consider implications for learning disability nursing, to increase awareness of research and policy issues, and ultimately to enhance practice. Preliminary findings suggest that participants have moved from an entry position of professional bias and lack of knowledge and skill, to a changed perspective and enhanced practice relating to the mental health needs of clients with a learning disability. These changes are, however, speculative and may be attributable to variables other than module completion. Implications for further research and development are suggested. PMID- 10693484 TI - Problem-based learning: an educational strategy to support nurses working in a multicultural community. AB - Physical dislocation of people from their homelands either as refugees, immigrants or exiles has resulted in the creation of multicultural communities which have diverse health needs. Like elsewhere, nurses in Australia have been faced with the challenge of responding to an ever-changing migrant population. A modified problem-based learning project was conducted in Melbourne to assist nurses to enhance their practice of caring for children and families of non English speaking backgrounds (NESB). Clinical nurses worked with the researchers to develop and trial problem-based educational packages. The packages were designed for use in the clinical areas and graduate nursing programs to assist nurses overcome the cultural and communication difficulties they experienced when caring for people of NESB. PMID- 10693485 TI - Facilitating learning in the community with lecturer-practitioner posts. AB - The purpose of this paper is to present a case study of the perspectives of their role of a group of community lecturer-practitioners and a community teacher, who referred to themselves as 'community facilitators'. A qualitative design was used and data were collected by semi-structured interviews. These were transcribed and content analysis was undertaken. All participants provided a liaison role between the college and the community practitioners, prepared students for their clinical experience and assisted in relating theory to practice. The participants described how they managed the role and how they supported each other. Developing small teams of facilitators may provide a bridge between teachers and practitioners and can serve as a basis for further study of the lecturer practitioner role. PMID- 10693486 TI - Learning spiritual dimensions of care from a historical perspective. AB - The purpose of this article is to bring to focus an historical perspective to the subject of spirituality in nurse education. In doing so, the historical roots of spirituality in nursing are traced and commented. Whilst acknowledging the emerging perspectives on spirituality (Simsen 1986, Burnard 1986, 1987, Narayanasamy 1991, 1993, Harrison 1993, Bradshaw 1994, Ross 1995, Oldnall 1996, McSherry & Draper 1998) this paper attempts to address its historical dimension, which is presently lacking in the nursing literature. In order to address this historical gap in spirituality, this paper begins by looking at the spiritual influences of nursing in ancient civilizations like Egypt, Mesopotamia, China, Palestine, India, Greece and Rome and then examines the influence of Christianity. After this, the spiritual dimension of nursing is portrayed as it was in the Middle Ages, Renaissance, Eighteenth and Nineteenth centuries. Finally, the emerging nursing theories and their positions on spirituality (including those of humanists) are reviewed and commented. It is hoped that this paper, through a brief review of events, has begun to highlight the significance of the precursor to spirituality in nursing from an historical perspective. It is concluded that contemporary literature suggests there is scope for development of educational programmes to better equip nurses to meet patients' spiritual needs. PMID- 10693487 TI - The development of NEdSERV: quantitative instrumentation to measure service quality in nurse education. AB - The political climate of health care provision and education for health care in the latter years of the 20th century is evolving from the uncertainty of newly created markets to a more clearly focused culture of collaboration, dissemination of good practice, with an increased emphasis on quality provision and its measurement. The need for provider units to prove and improve efficiency and effectiveness through evidence-based quality strategies in order to stay firmly in the market place has never been more necessary. The measurement of customer expectations and perceptions of delivered service quality is widely utilized as a basis for customer retention and business growth in both commercial and non profit organizations. This paper describes the methodological development of NEdSERV--quantitative instrumentation designed to measure and respond to ongoing stakeholder expectations and perceptions of delivered service quality within nurse education. PMID- 10693488 TI - An exploratory study of nursing and medical students health promotion counselling self-efficacy. AB - Self-efficacy of nursing and medical students for client health promotion counselling was examined in an exploratory study using Bandura's (1977) self efficacy theoretical perspective. Third-year nursing students (n = 41) and fourth year medical students (n = 60) were compared on their self-efficacy for engaging in clients health promotion activities within 5 areas: smoking, exercise, nutrition, sexually-transmitted diseases and injuries. Their self-efficacy about their knowledge levels in the same 5 areas of health was also compared along with their perceptions of the relative impact of various curricular learning experiences on building health promotion counselling self efficacy. Self-efficacy scores were high for both groups. Nursing students scored significantly higher on both knowledge and ability to counsel patients in the areas of exercise, nutrition and injury prevention. In both groups, confidence in knowledge for health promotion activities was higher than ability to counsel patients. Learning specific health promotion strategies in class and actual practice were strongly associated with nursing students' self-efficacy while practice, feedback on performance, and role modelling were strongest for medical students. PMID- 10693489 TI - Achievement motivation, anxiety and academic success in first year Master of Nursing students. AB - Forty-one first semester master level nursing students from three Canadian universities participated in this descriptive correlational study to identify the influence of achievement motivation and anxiety on their academic success. Academic success was determined by their first semester grade point average (GPA). Participants had high achieving tendencies (M = 73.5) and academic ability (M = 81.9), supporting Atkinson's (1957, 1964) achievement motivation theory which was used as the framework. While state anxiety was negatively correlated, trait anxiety was the only valid predictor of academic success. Academic ability and inherent anxiety had a greater potential for predicting students who would succeed, which has implications for nurse educators, administrators and researchers. However, the need to assess both cognitive and non-cognitive variables to determine master level nursing students' ability to succeed is recommended. PMID- 10693490 TI - Learning and teaching beyond 2000. PMID- 10693491 TI - Insufficient evidence: the problems of evidence-based nursing. AB - Evidence-based medicine was first proposed in the early 1990s as a means of integrating clinical expertise with the best evidence from research. It has recently gained a foothold in nursing, where despite calls for a broad and nursing-oriented definition of what should count as evidence, it appears to be propounding the randomized controlled trial (RCT) as the gold standard. This paper challenges the wisdom of basing nursing practice on the findings of large scale statistical research studies, and offers a number of logical objections to the underpinning philosophy of evidence-based nursing and the randomized controlled trial. It concludes by arguing for a rethinking of what should count as evidence, and suggests a quasi-legal model based on reflection rather than research, in which evidence is employed to understand and justify practice after the event rather than being used deterministically to plan practice in advance. PMID- 10693492 TI - Students' part-time work: towards an understanding of the implications for nurse education. AB - This paper reports on the subject of nursing students undertaking paid part-time work, an area which has attracted minimal research. A questionnaire survey of 120 diploma students in one university investigated several aspects of part-time work: incidence, nature, motivating factors, and effects on their professional and personal lives. The results provide an insight into the work profile and the students' perceptions of the positive and negative effects on their personal well being and progression on the course. The findings raise issues which are of interest to both academic and clinical staff, nursing students and employers. PMID- 10693493 TI - Nurses' perceptions of the value of written reflection. AB - Reflective writing is increasingly becoming a feature of professional practice as nurses seek to provide evidence of their continuing development and competence. This study reports the process and findings of a study using grounded theory to explore how nurses are using and developing writing techniques as a tool for facilitating and supporting their development in practice. Two focus group interviews were conducted with 12 experienced nurses completing a professional course which involved reflective writing as the assessment component over a calendar year. These interviews generated the broad base of concepts and categories which direct the later stages of a grounded theory study. These preliminary categories suggest that firstly, the skills of reflective writing need to be learnt rather than being assumed as a natural capacity; secondly, that this leads to the acceptance of writing as a learning strategy in its own right; thirdly, reflective writing is considered to be a tool which helps the practitioner to develop analytical and critical abilities; finally, the nurses identified their own personal, as well as professional growth as being facilitated by reflective writing. Lincoln and Guba's (1985) criteria for establishing rigour in qualitative studies, and Strauss and Corbin's (1990) criteria for judging a grounded theory study are used as benchmarks throughout the paper. PMID- 10693494 TI - Attitudes of Hong Kong high school students towards the nursing profession. AB - The increased development of hospital services in Hong Kong over the last decade has given rise in the demand for more recruits to join the nursing profession. Despite the advancement in education and the improvement in the working conditions, the problems of attracting sufficient new recruits remain critical. This study aimed to examine high school students' attitudes towards the nursing profession and to identify the contributing factors affecting shortage of nurses within the context of Hong Kong. A convenience sample of 375 high school students was recruited. A questionnaire was used to measure their knowledge, attitudes and intention to study nursing. Descriptive and inferential statistics were used to describe their career preferences and to compare knowledge, attitudes and intention scales between gender and nursing exposure groups. Results indicated that the students were generally knowledgeable about nursing but were reluctant to pursue nursing as a career. However, students who were socially acquainted with a nurse demonstrated a slightly more positive attitude towards nursing and slightly higher intention to pursue nursing as a career compared with those having no social acquaintance with a nurse. Implications for promotion of nursing profession and limitations of the study were discussed. PMID- 10693495 TI - Change of outlook on elderly persons with dementia: a study of trainees during a year of special education. AB - In 1996 the Silvia Home Foundation started a non-governmental education programme with an integrated day-care unit devoted to patients with dementia: Working chairwoman of the foundation is HM Queen Silvia of Sweden. This study aims to describe the trainees' changed outlook on elderly people with dementia during a year of special education. Data were collected by interviews, informal discussions and participant observations during the lessons. The investigation focused on two questions: the trainees' outlook on the patients and their outlook on the work with the patients. At the beginning of their education, the trainees looked at the patients from a staff's perspective. During their education, this was gradually toned down and they changed to a disease perspective, and eventually to a human dignity perspective. After initially seeing dementia patients as a homogeneous group, the trainees went on to see them as unique human beings. Their outlook on their work also changed, from being task-oriented to a more humanitarian approach. PMID- 10693496 TI - Transfer of knowledge and skills: some implications for nursing and nurse education. AB - The construct of transfer has enormous importance to nursing as it begins to highlight potential problems in the transfer of knowledge and skills from the campus to the clinical area, from one part of the clinical area to another (e.g. surgical to medical), and from community to the clinical area. Thus, any adequate conceptualization of transfer must account for problems of practice-practice transfer as well as theory-practice transfer. These potential problems are the concern of educators, students and managers who have a responsibility for agency nurses and bank nurses who may find themselves in different specialities on a regular basis. Transfer has relevance to a whole raft of other issues ranging from the application of theories to nursing practice, through to the validity of claims that courses which develop intellectual skills prepare nurses for lifelong learning. PMID- 10693497 TI - Developing a distance learning honours degree in health promotion for delivery using the Internet. AB - Designing this programme for distance learning delivery, especially for the Internet has been a major learning experience for all the staff involved. However, addressing and resolving these issues has now provided us with a template for developing other programmes in distance learning mode. This was an endeavour that all our market research indicated we had to undertake since institutions of higher education, especially schools of healthcare are no different to commercial companies. Like commercial organizations, higher education institutions, too, are consumer facing and are in direct competition with other educational 'suppliers', i.e. other universities and colleges. If a university is to compete and to 'sell' its products well, it too must consider the Internet as a way of advertising its wares, streamlining its business model and eventually conducting its business. In our case, this means delivery of educational programmes. However, as Internet technologies are primarily designed for research and business, there is currently a lag in the development of tools that enable effective social learning. Our next programme via the Internet will be even better for the experiences we've undergone and the lessons we've learnt during this initiative. PMID- 10693498 TI - Delphi technique: one cycle of an action research project to improve the pre registration midwifery curriculum. AB - Facilitating a unified interpretation of new or re-designed curricula can be problematic in multi-sited universities. This paper discusses the use of the Delphi technique when implementing an innovative pre-registration midwifery curriculum. It formed one of the action research cycles of the author's doctoral study and was used in an attempt to seek the views of all midwife teachers and achieve a critical mass consensus. The most important issues that emerged from the Delphi process were grouped into four categories. These four categories (the teacher's base site, communications, staff development and administration issues) formed the agenda for a staff away day. The outcome of the process was the generation of a list of priorities and actions for curriculum implementation and the identification of ways in which staff development needs might be met more effectively. An unexpected, but equally important, outcome was the way it helped with team building and agreement on positive actions for its further development. PMID- 10693499 TI - Parenting by people with learning disabilities: the educational needs of the community nurse. AB - This article discusses some of the findings of a research project which was designed to examine the role of community learning disability nurses in supporting people with learning disabilities who become parents. The need for information and help with the experience of pregnancy, childbirth, becoming parents and bringing up children has been highlighted in a recently issued guide for commissioning and providing services for people with learning disabilities (NHS Executive 1998). This research, however, indicates that nurses do not feel themselves to be adequately prepared for their role in supporting parenting. Responses from a questionnaire survey of 266 practitioners revealed little coverage of key issues relating to parenting on either pre-registration or post basic courses and suggests, therefore, that some courses may not be meeting the perceived needs of these nurses in this crucial aspect of their role. This is likely to have significant consequences for this client group and suggests that educationalists need to consider ways of including more extensive discussion of parenting in educational provision for the multiplicity of professional groups who support and care for adults with learning disabilities. PMID- 10693500 TI - Specialist practice in community mental health nursing. AB - Community mental health nurses (CMHNs) work in an increasingly complex health and social care environment. Over recent years, the evolving direction of general health service and specific mental health policy has directed CMHNs towards: the provision of clinically-effective interventions; a closer attention to meeting the needs of people experiencing severe and long-term mental health problems; the simultaneous provision of services to meet the needs of people experiencing a wide range of mental health problems presenting in primary care settings; greater collaboration with workers representing other disciplines and agencies; and the development of active partnerships with mental health service users. This paper explores the context within which CMHNs practise, and within which education programmes preparing specialist practitioners in community mental health nursing have been developed. One recently-validated specialist practice course for CMHNs is described in detail, with the intention of stimulating discussion and debate surrounding the practice of, and the educational preparation for, community mental health nursing. PMID- 10693501 TI - UKCC Education Commission. A little fine tuning. PMID- 10693502 TI - Party season. PMID- 10693503 TI - Unfinished business. Interview by Charlotte Alderman. PMID- 10693504 TI - Nature's prescription. PMID- 10693505 TI - Smart moves. Interview by Dina Leifer. PMID- 10693506 TI - Fighting the flu. PMID- 10693507 TI - The painful truth. PMID- 10693508 TI - Violence: a worldwide epidemic. AB - Nurses are suffering from society's acceptance of violence worldwide, says the International Council of Nurses. What is needed is a campaign of zero tolerance. PMID- 10693509 TI - Laxatives and faecal incontinence in long-term care. AB - High levels of faecal incontinence and laxative use in long-term care settings for older people are revealed in this study. Further research, together with more considered prescribing policies are needed, the authors suggest. PMID- 10693510 TI - Investing in continence. AB - Services for people with incontinence are dependent on location and available resources. Sue Thomas urges nurses to be proactive in lobbying for more equity in continence care. PMID- 10693511 TI - Keeping the lines open. AB - Central venous catheters (CVCs) often become occluded. Many of the guidelines for the use of CVCs are ambiguous and do not offer clear guidelines for practitioners. This article discusses the development of a local policy for maintaining or increasing the patency of CVCs in acutely ill adults. PMID- 10693512 TI - The benefits of hypodermoclysis. AB - Hypodermoclysis (subcutaneous fluid administration) is a valuable means of replacing fluids and maintaining hydration for older people and palliative care patients. This literature review looks at three aspects of the therapy--client groups, wound care and types of infusion fluids. PMID- 10693513 TI - Assertiveness, power and influence. AB - This article describes how nurses can use the concepts of assertiveness, power and influence to maximise their effectiveness in delivering care and bringing about change. PMID- 10693515 TI - Learning disability nursing--holistic care at its best. PMID- 10693514 TI - Giving injections. PMID- 10693516 TI - A balance of power. PMID- 10693517 TI - [We will have to work together]. PMID- 10693518 TI - [Accreditation, a landmark]. PMID- 10693519 TI - [The process of accreditation from the viewpoint of the nursing service]. PMID- 10693520 TI - [Accreditation. The Canadian model, the medical viewpoint]. PMID- 10693521 TI - [Accreditation. A tool to serve the improvement of quality]. PMID- 10693522 TI - [Accreditation. An admission strategy, a means of patient-centered care]. PMID- 10693523 TI - [Accreditation. Participation of the consumers in the evaluation of satisfaction]. PMID- 10693524 TI - [Therapeutic impasse. The capacity to make everything fail]. PMID- 10693526 TI - [Surfing in nursing care]. PMID- 10693525 TI - [Short term therapy, another look at the change]. PMID- 10693528 TI - [The National Network of Hospital Documentors, a network to be discovered] [In Process Citation] PMID- 10693527 TI - [The birth of Nursing Convergence upsets the French nursing syndical scene] [In Process Citation] PMID- 10693529 TI - [A new decree for a unique competency] [In Process Citation] PMID- 10693530 TI - [Finding a manageable distance]. PMID- 10693531 TI - [Sexual delinquency, reference points]. PMID- 10693532 TI - [The law of June 17, 1998]. PMID- 10693533 TI - [Respecting every type]. PMID- 10693534 TI - [The judge pronouncing the fine, focus of application. Interview by Francois Hamon]. PMID- 10693535 TI - [Incarceration of sex delinquents. Interview by Francois Hamon]. PMID- 10693536 TI - [The pervert, the law and the therapist]. PMID- 10693537 TI - [Approach to the care of a pedophile]. PMID- 10693538 TI - [Psychotherapies of sex delinquents]. PMID- 10693539 TI - [1999, psychiatry of the stone age]. PMID- 10693540 TI - [A disarmed nursing team]. PMID- 10693542 TI - [We need a law to develop palliative care] PMID- 10693541 TI - [Surfing in nursing care]. PMID- 10693543 TI - [Rare diseases and the health care system, 1. citizens' forum] [In Process Citation] PMID- 10693544 TI - Ultrasonography as adjunct to mammography in the evaluation of breast tumors. AB - PURPOSE: The aims of the study was to analyze the features of cancers missed as tumor on ultrasonography (US), to determine the predictive power of US tumor descriptors in the differentiation of benign and malignant breast tumors, to evaluate US as adjunct to mammography, and to assess the validity and reliability of mammographic, US, and combined interpretation of breast tumors. MATERIAL AND METHODS: Prospectively recorded mammographic and US findings of 355 malignant breast tumors among 2,985 consecutive patients who underwent breast US were compared with clinical findings and pathologic subtypes of the tumors. In addition, a 10-year material of 95 invasive lobular carcinomas (ILCs) were investigated. Three retrospective studies assessed the validity and reliability of mammographic, US, and combined interpretation of 200 palpable mammographically noncalcified breast masses by four radiologists. RESULTS: A total of 97.5% of the palpable and 67.9% of the nonpalpable malignant neoplasms were detected as tumor on US. Most carcinomas missed as tumor on US were ductal carcinomas in situ (DCISs) and microinvasive ductal cancers dominated by DCIS manifesting with suspicious calcifications on mammography. Irregular shape, irregular contour, extensively hypoechogenicity, hyperechoic rim (halo), and distortion of the surrounding tissue were the US features with the highest odds of predicting carcinomas. A pseudocapsule is the strongest predictor of a benign tumor, the odds of cancer being 0.03 in nonpalpable and 0.08 in palpable breast tumors. A negative predictive value of 100% in palpable and 96% in nonpalpable tumors was achieved using strict US criteria. In patients with ILC, US measurements predicted tumor size more accurately than mammography. US as adjunct to mammography correctly diagnosed ("upgraded") 9.5% of tumors with benign or indeterminate mammographic diagnoses. Excluding mammographically conclusive malignant tumors and carcinomas presenting with microcalcifications. US correctly upgraded 42% of the palpable and 44% of the nonpalpable cancers. Combined mammographic-US interpretation offers the highest diagnostic performance in noncalcified breast tumors. The lowest interobserver agreement was found in US interpretation. CONCLUSION: The impact of US in mixed cancer populations is limited. US is, however, a valuable adjunct to mammography in patients with nonconclusive mammographic findings. Negative predictive values on US approaching 100% may be achieved using strict criteria for a benign diagnosis. A considerable interobserver variation in the US interpretation is a limiting factor for the potential of breast US in the differentiation of benign and malignant breast tumors. PMID- 10693545 TI - Transplantation of tissue from lower animals to man, and a report of the case of bone-transplantation at Charity Hospital, Blackwell's Island, N.Y. 1891. PMID- 10693546 TI - Bone graft materials. An overview of the basic science. AB - Autograft, allograft, and synthetic bone graft substitute materials play an important role in reconstructive orthopaedic surgery, and understanding the biologic effects of these materials is necessary for optimum use. Although vascularized and cancellous autograft show optimum skeletal incorporation, host morbidity limits autograft availability. Experimental studies have confirmed an immune response to allograft bone, but the clinical significance of this response in humans still is unclear. Small amounts of cancellous allograft in humans usually are remodeled completely; large allografts become incorporated by limited, surface intramembranous bone formation suggesting that these graft are primarily osteoconductive. Several synthetic skeletal substitute materials also are osteoconductive, and may show remodeling characteristics similar to allograft. Demineralized bone matrix and some isolated or synthetic proteins can induce endochondral bone formation, and therefore are osteoinductive. The extent and distribution of remodeling of bone graft materials are influenced by many factors, including the quality of the host site and the local mechanical environment (strain). Graft materials are likely to become more specialized for use in specific clinical applications, and composite preparations may soon provide bone graft materials with efficacy that equals or exceeds that of autogenous grafts. PMID- 10693547 TI - Posterolateral and anterior interbody spinal fusion models in the sheep. AB - Posterolateral and anterior interbody spinal arthrodesis is a frequent procedure, but high nonunion rates are reported and harvesting autologous bone graft from the iliac crest significantly increases morbidity. Bone graft substitutes are an alternative, but to date clinical results are not conclusive. Bone substitutes can be organic or inorganic, biologic or synthetic. They can have osteoconductive properties, inductive properties or both. Animal experiments are essential to investigating bone substitutes using biomechanical and histologic methods not available in clinical studies. Few authors reported on instrumented anterior fusion models, but none used the sheep model. In the current study posterolateral and anterior interbody fusion models in sheep are described. Both models used instrumented fusions, applying porous mineral scaffolds, alone or mixed with bone. The surgical techniques are described step-by-step and potential difficulties are highlighted. Preliminary results are reported for the posterolateral fusion model using coralline graft substitutes. The coral granules mixed with locally harvested bone had fusion outcomes similar to pure autologous bone. The graft substitute showed marked resorption between 12 and 20 weeks. All fusions had bone cortex and good trabecular connectivity. Histologic evaluation suggests after 20 weeks nearly the entire surface of the substitute is covered with new bone. Porous mineral bone substitutes mixed with locally harvested autologous bone are thought to be a valid alternative for posterolateral fusions. PMID- 10693548 TI - The use of allograft bone in lumbar spine surgery. AB - Bone grafting is an integral part of many lumbar spinal surgeries. The two choices of bone are autograft and allograft. Each source has its own advantages and disadvantages. The current study is a literature review of allograft bone use in lumbar spine surgery. Allograft bone can be procured in greater quantities than autograft. With standard protocols of harvesting, the risk of disease transfer is negligible. Only fresh-frozen and freeze-dried products are used. Allografts are incorporated slower and to a lesser degree than autografts. Fresh frozen grafts are stronger, more immunogenic and more completely incorporated than freeze-dried grafts. Allografts used alone or combined with autografts for posterior lumbar spinal procedures have decreased fusion rates compared with autografts. If used anteriorly, allografts are well suited for reconstructive procedures and have good fusion rates, especially if combined with posterior fusions. If used in the proper situations, allograft bone can be used successfully in lumbar spine surgeries. PMID- 10693549 TI - Pseudarthrosis repair. Autogenous iliac crest versus femoral ring allograft. AB - Pseudarthrosis repair in the lumbar spine is one of the most challenging problems faced by spine surgeons. Historically high failure rates with posterior repair have led to the use of anterior lumbar interbody fusion with tricortical iliac crest autograft in these difficult cases. More recently, femoral ring allografts packed with autograft bone have been advocated as another method that would decrease donor site morbidity. Two series of patients underwent anterior lumbar interbody fusion with anterior instrumentation to repair pseudarthrosis (Group I, 33 patients with tricortical autogenous iliac crest and Group II, 20 patients with femoral ring allografts). At minimum 2-year followup, there was no difference in fusion rates (Group I, 32 of 33 versus Group II, 20 of 20). Patients in Group I had radiographic fusion develop more rapidly than patients in Group II (12 months versus 18 months), but a significant proportion of patients in Group I (35%) had an average of 2 mm of graft subsidence. Despite excellent fusion rates in both groups, functional outcomes were not as good with only 28% of patients in Group I and 36% of patients in Group II returning to work. Using anterior instrumentation, anterior interbody fusion offers an excellent method to repair pseudarthrosis using femoral ring allografts or autogenous iliac crest. However, femoral ring allografts offer the potential to decrease donor site morbidity, allowing the surgeon to treat multiple spine levels. PMID- 10693550 TI - Anterior lumbar interbody fusion with osteoinductive growth factors. AB - Anterior intervertebral fusion increasingly is used as a treatment for discogenic or intersegmental pathologic diseases of the lumbar spine. This is in part attributable to the evolution and refinement of laparoscopic and minimally invasive surgical techniques that now can be used to access the anterior spinal column. It also is attributable to the availability of newer generation intervertebral fixation devices such as the threaded titanium cages or threaded allograft bone dowels, both of which are technically simpler to implant. Recently, limited clinical studies of intervertebral lumbar fusion have examined the use of these devices combined with osteoinductive growth factors as substitutes for autogenous bone graft. Early clinical results of lumbar fusion using threaded intervertebral implants filled with recombinant human bone morphogenetic protein-2 have been favorable. Higher fusion rates, shorter operative times, and shorter hospital stays have been reported in the initial series. Clinical trials involving larger cohorts with various spinal applications for osteoinductive molecules currently are in progress. PMID- 10693551 TI - Human bone morphogenetic protein allografting for reconstruction of femoral nonunion. AB - A composite inductive allograft consisting of an allogeneic, autolysed, antigen free cortical bone carrier lyophilized with partially purified human bone morphogenetic protein was implanted in 30 consecutive femoral reconstructions that resulted from failure of fracture healing. There were 24 atrophic shortened femoral nonunions, four equal length femoral nonunions, and two femoral malunions. There were 10 men and 20 women with an average age of 47 years (range, 28-75 years). Allogeneic, autolysed antigen-free cortical bone was used as a structural alloimplant and as a delivery system for partially purified human bone morphogenetic protein. The composite implant of human bone morphogenetic protein/allogeneic, autolysed antigen-free cortical bone was used in conjunction with one-stage lengthening of the extremity, restoration of mechanical axis and rotational alignment. In 26 of 30 femurs, the human bone morphogenetic protein/allogeneic autolysed antigen-free cortical bone consisted of an allogeneic cortical bone implant incorporated into a one-stage lengthening of atrophic femoral nonunion. In four patients with equal length femoral nonunions, the human bone morphogenetic protein/allogeneic, autolysed antigen-free implant was placed as an medical femoral shaft onlay graft. Internal remodeling of the implant occurred within 8 to 12 weeks after implantation. Lengthening defects greater than 2 cm were supplemented with intercalary autogeneic bone graft. Twenty-four femurs healed at an average of 6 months at an average followup of 55 months. Four of six plate fatigue failures were salvaged with repeat plating. Two patients were lost to followup. The human bone morphogenetic protein/allogeneic, autolysed antigen-free bone allograft is an excellent structural and delivery system that induces host bone formation and implant remodeling allowing salvage of difficult femoral nonunions. PMID- 10693552 TI - Minor column structural acetabular allografts in revision hip arthroplasty. AB - A minor column (shelf) allograft is used for uncontained defects that involve less than 50% of the acetabulum. The prospectively collected records and radiographs of 47 patients (51 hips) who had undergone minor column structural acetabular allograft reconstruction during revision hip arthroplasty were reviewed. The purpose was to identify the long-term results (minimum 5 years) and factors that may influence longevity of the allograft and predispose the patient to subsequent acetabular component failure. The mean duration of followup was 119 months (range, 68-195 months). There was one perioperative death and six patients were lost to followup. Eleven patients (22%) required additional surgery. Three acetabular cups could not be revised successfully, despite multiple attempts, and the patients were treated with Girdlestone excisions. Eight patients underwent successful revision surgery with only three requiring a repeat structural allograft. Survival time for the acetabular cup as determined by Kaplan-Meier analysis was 153 months (95% confidence interval; range, 136-169 months). Cup failure was associated with more operative procedures performed before revision surgery (mean, 3.2 procedures), and failure to restore the vertical center of hip rotation to within 12 to 14 mm of the predicted value. The acetabular abduction angle was not a predictor for failure. The current study shows that good results can be achieved with structural acetabular allograft reconstruction with mid-term to long-term implant survival (cup aseptic survival, 80.4% and allograft re construction survival, 94.1%), especially if there is restoration of near normal hip biomechanics. PMID- 10693553 TI - Cancellous allograft in revision total hip arthroplasty. A clinical review. AB - There are numerous indications for the use of cancellous allograft bone in the context of revision hip arthroplasty. These indications range from the well documented use of morselized bone chips to fill cavitary defects during cementless acetabular reconstructions--in which complete or near-complete graft remodeling is expected--to the use of particulate allografting with bipolar hemiarthroplasty for acetabular revision, which largely has been abandoned because of frequent component migration, graft resorption, and clinical failure. Most other indications, including femoral and acetabular impaction allografting techniques, curettage of osteolytic defects with component retention, and complex reconstructions using acetabular reconstruction rings or cages with cancellous donor bone, are controversial but are supported by published clinical series. The current study reviews the literature on cancellous allografting in revision total hip arthroplasty. PMID- 10693554 TI - Circumferential allograft replacement of the proximal femur. A critical analysis. AB - The use of proximal femoral structural allografts in revision hip arthroplasty remains controversial. The current study constitutes the mean 8.8 years followup (range, 3-12.5 years) of a consecutive series of 55 proximal femoral allografts in 51 patients. In 46 patients the implant was cemented into the allograft and the distal femur, and the host proximal femur was resected at the time of reconstruction in all but seven patients. Five patients underwent revision surgery for acetabular failure, and six additional patients underwent revision surgery for failure of the proximal femoral allograft. Three patients underwent successful revision surgery and had additional proximal femoral allografts. Failure was caused by graft fracture in one patient, by deep infection in two patients, and by junctional nonunion in three patients. Junctional nonunion was seen in five patients (9%), two of whom were treated successfully with bone grafting and bone grafting and plating, respectively. Instability was observed in six patients (11%). Trochanteric nonunion was seen in 22 patients (43%) and trochanteric escape was seen in 14 patients (27%). The mean Harris hip score improved from 39 to 79 points. Resorption involving the full thickness of the allograft in at least one zone was seen in seven patients. This progressed rapidly and silently within the first 3 years but has yet to lead to the failure of any of the reconstructions. Infection was ruled out in every case. Allograft resorption was seen in seven patients and may be related to a combination of factors. It is most likely that this is an immunologic problem of slow rejection, but it is possible that the distal cement fixation led to stress shielding and resorption attributable to mechanical disuse. The possible protective role of retaining the bivalved host bone as a vascularized onlay autograft remains to be clarified. Although these results justify the continued use of structural allografts for selected patients, continued followup is warranted. PMID- 10693555 TI - Results of onlay allografts. AB - Cortical onlay strut allografts provide a method for bone restoration when performing revision surgery in a patient with a structurally deficient femur. Between 1986 and 1990, 251 patients underwent femoral revisions using structural onlay bone grafts. The followup ranged between 8 and 12 years, with the average followup being 9.5 years. All of the grafts united to the host bone. The revision rate in the current series is 3%, with no complications related to the bone graft. The average Harris hip score improved 45 points. Cortical onlay grafts are used for patients with structurally deficient femurs. After union occurs, the graft undergoes adaptive remodeling, secondary to physiologic load bearing. This technique has proved to be a reliable method for bone restoration in the patient with a structurally compromised femur. PMID- 10693556 TI - Revision total knee arthroplasty by impaction bone grafting. AB - The presence of bone loss in a failed total knee arthroplasty can present a significant reconstructive challenge. Experience with the technique of using impacted morselized allograft with revision components having fixed stems is presented. Nineteen knees (21 patients) were reconstructed using impacted bone graft alone in 14 knees, bone graft plus methylmethacrylate in five knees (including one knee in which the replacement failed), and in three knees morselized bone graft was used in conjunction with structural bone allograft. Minimum followup ranged from 6 months to 62 months for the patients in the current series. These patients represent a relatively small, but growing portion of this surgeon's population of patients undergoing revision knee arthroplasty. Patients with large defects were selected for the study. Histologic specimens from the one failed knee arthroplasty revealed viable, incorporated bone graft. Excluding the replacement that failed, the average improvement in Knee Society combined knee and function scores was 87 points. The principles of revision and primary total joint arthroplasty are applied for achieving a stable implant. Specific to this technique, solid support of the implant-graft interface, graft host bone interface, and the use of a tight, supportive stem is imperative. The author's experience provides additional support for the use of bone grafting techniques in patients with large bone defects who are undergoing revision total knee replacement. PMID- 10693557 TI - Surgical management of Sprengel's deformity in adults. A report of two cases. AB - Sprengel's deformity is a congenital structural abnormality of the shoulder girdle, and most patients with Sprengel's deformity receive surgical treatment as a child or adolescent. Thus, it is uncommon to identify an adult with untreated Sprengel's deformity. Two adult patients who had no previous surgical treatment were treated with resection of an omovertebral bone and resection of the superomedial border of the scapula. This procedure was not overly invasive or technically demanding. In both patients resection of the omovertebral bone provided an improved cosmetic appearance and reduction in neck pain, although shoulder motion was improved in only one patient. Because the patient with a long omovertebral bone adjacent to the levator scapulae muscle showed great improvement in her shoulder motion, the size, shape, and position of the omovertebral bone may predict improvement of shoulder motion after resection. Thus, the surgical treatment of adults patients with Sprengel's deformity can produce good surgical results. PMID- 10693558 TI - Successful restoration of the trapezius muscle using pedicle latissimus dorsi. A case report. AB - Reconstruction of the trapezius muscle using a pedicle latissimus dorsi flap was performed in a 27-year-old man with a large synovial sarcoma in his shoulder girdle. Size and location of the tumor required combined resection of surrounding muscles, including the trapezius, levator scapulae, and rhomboid major and minor. Thus, an extensive defect of the suspending muscles of the scapula was created after accomplishing an adequate resection of the tumor. The flap was performed to restore the trapezius functionally because there were no adjacent muscles available. The transferred muscle compensated for loss of the trapezius, thereby recovering excellent shoulder function. Although an opportunity of its application is thought to occur infrequently, the pedicle latissimus dorsi can activate scapular motion successfully in the absence of the levator scapulae. The technique may be extended to salvage failed conventional reconstruction after spinal accessory nerve palsy. PMID- 10693559 TI - Palmar approach in flexible implant arthroplasty of the proximal interphalangeal joint. AB - Joint replacement is an established method in the treatment of destroyed, painful, proximal interphalangeal joints. A palmar approach was used in which the main collateral ligaments were preserved, allowing immediate active rehabilitation with enhanced primary lateral stability. Fifty-nine proximal interphalangeal joint silicone arthroplasties in 38 patients with a minimum followup of 12 months were reviewed. Thirty-eight of the 59 joints had implantation from a palmar approach and 21 joints from a dorsal approach. The two groups were well-matched in terms of indication, preoperative range of motion, and patient age. No significant increase in the range of motion was found in either of the patient groups, with an overall average range of motion of 51 degrees postoperatively. There was also no significant difference in the postoperative stability in the two patient groups. The choice of surgical approach at the proximal interphalangeal joint level for the silastic type of implants does not appear to be important. With more sophisticated types of implants in which the integrity of the collateral ligaments is crucial, a palmar approach might be beneficial. PMID- 10693560 TI - Hip and knee replacement after longstanding hip arthrodesis. AB - This study determined whether patients with severe knee disease below a hip arthrodesis can be treated successfully with total knee replacement alone or whether such patients require total hip arthroplasty followed by knee replacement. Eighteen patients who had hip arthrodesis for a mean of 33 years underwent total hip replacement alone, total knee replacement alone, or a combination of both. The Harris hip score improved from a mean of 55.3 to a mean of 86.9 points at 45 months after total hip arthroplasty. The Hospital for Special Surgery knee score improved from a mean of 33 to a mean of 78 points in patients who had total knee replacement after total hip arthroplasty. The Hospital for Special Surgery knee score improved from a mean of 35 to a mean of 44 points in patients having total knee replacement alone below a hip arthrodesis. The followup after total knee replacement averaged 53 months. These data suggest that a knee replacement alone in a patient with a fused hip is unlikely to provide a satisfactory result. Patients with severe knee disease below hip arthrodesis require total hip arthroplasty followed by knee replacement. This applies even when severe osteoarthritis of the knee is the primary complaint. PMID- 10693561 TI - Influence of porous coating level on proximal femoral remodeling. A postmortem analysis. AB - This study used femurs retrieved at autopsy to compare the extent and location of bone remodeling between four patients implanted with proximally porous coated femoral prostheses and a matched group of four patients implanted with extensively porous coated femoral prostheses. The femoral components studied were large, cementless, straight, cobalt chrome stems and were identical except for the amount of porous coating. The contralateral normal femur of each patient also was retrieved, implanted with an identical prosthesis, and used as a control for bone mineral content. Dual energy x-ray absorptiometric analysis showed marked loss of bone mineral content in both groups of patients. The extensively coated group had less bone loss on average (18.4%) than did the proximally coated group (38.6%). There was no relationship between the extent of coating and the location of bone mineral loss; specifically, proximal coating did not protect against loss of bone mineral content proximally or distally in the femur. Videodensitometric analysis of cross sections of periprosthetic bone also showed that the extensively coated group tended to have less decrease in bone density than did the proximally coated group (14.3% versus 28.4%). Although one cannot presume that all proximally fixed stem designs would produce results similar to those presented here, these findings show that decreasing the extent of porous coating alone does not necessarily reduce proximal femoral bone loss. PMID- 10693562 TI - Bilobed oblong porous coated acetabular components in revision total hip arthroplasty. AB - Thirty-eight oblong bilobed noncustom uncemented, porous-coated titanium acetabular components were used to reconstruct failed hip arthroplasties with large superior segmental acetabular bone deficiencies. No structural bone grafts were used. All patients were followed up for 2 to 5 years (mean, 3 years) after the operation. One patient (whose socket rested primarily on a structural bone graft from a previous procedure) had revision surgery for acetabular loosening. No other patients have had revision surgery or had another ipsilateral hip operation. At latest followup, 35 patients had no or mild pain and two patients had moderate pain. Two implants migrated more than 2 mm in the first year, then stabilized. On the latest radiographs, two implants had bead shedding, but there was no measurable migration or change in position. For selected patients with large superolateral acetabular bone deficiencies, this implant facilitated a complex reconstruction, provided good clinical results, and showed satisfactory stability at early to midterm followup in most patients. PMID- 10693563 TI - Subvastus and medial parapatellar approaches in total knee arthroplasty. AB - This retrospective study compared the outcome of two consecutive groups of patients having primary total knee arthroplasty. The arthroplasties were performed in the first group (169 arthroplasties in 143 patients) from 1988 to 1992 using a medial parapatellar approach, and in the second group (167 arthroplasties in 148 patients) from 1992 to 1996 using a subvastus approach. The patient outcomes were evaluated at 6 months, and were based on clinical and radiographic measures, occurrence of intraoperative lateral retinacular release, and incidence of postoperative patellar subluxation. There were no significant differences between the two groups for range of motion, Knee Society knee and function scores, and stair climbing ability. The patella tracked centrally in significantly more knees with the subvastus approach (139 of 167 knees, 83%) than with the parapatellar approach (107 of 169 knees, 63%). There were significantly fewer knees in the subvastus group requiring a lateral retinacular release (62 of 167 knees, 37%), compared with the parapatellar group (113 of 169 knees, 67%). The authors concluded that the subvastus approach led to improved patellar tracking and stability. Although the surgical and rehabilitative protocols were identical for both groups, the results may have been affected by changing circumstances during the 9-year period of the study. PMID- 10693564 TI - Effect of tourniquet use on activation of coagulation in total knee replacement. AB - Total knee replacement often is performed with tourniquet application. The advantages of a dry field, including fixation, are well known, but it still is debatable if tourniquet application increases deep vein thrombosis. Measurement of coagulation markers is a well accepted method of studying thrombogenesis activation intraoperatively and postoperatively. Twenty patients undergoing total knee replacement with subarachnoid anesthesia were assigned randomly to two groups: either with tourniquet application (Group I) or without tourniquet application (Group II). There were no differences between patients in the two groups in terms of age, gender, diagnosis (all had osteoarthritis), operative time, and total (intraoperative and postoperative) blood loss. Markers for thrombin generation and fibrinolysis were measured. Blood samples were drawn at four times: baseline before the operation; after bone cuts; after cement fixation (Group II) or 2 minutes after tourniquet deflation (Group I); and 1 hour after surgery. Markers of thrombin generation and fibrinolysis showed a significant increase from baseline in all the patients. In Group II these markers started to increase during surgery, whereas in Group I the increase occurred at the end of the procedure when the tourniquet was deflated. The total amount of thrombin generation was significantly higher in Group II (without tourniquet), whereas fibrinolysis was significantly greater in Group I. Total knee replacement is accompanied by a hypercoagulative state with or without the use of a tourniquet, but it seems to be higher when the tourniquet is not used. In addition, tourniquet application may increase fibrinolysis. PMID- 10693565 TI - Axis location of tibial rotation and its change with flexion angle. AB - The magnitude and location of the axis of tibial rotation were measured at 15 degrees increments between 0 degree and 90 degrees flexion using 24 normal anatomic knee specimens, and their changes with flexion angle were investigated. The magnitude of tibial rotation was small (8.3 degrees) at 0 degree flexion, but increased rapidly as the flexion angle increased and reached a maximum rotation (31.7 degrees) at 30 degrees flexion. It then decreased again with additional flexion (24.8 degrees at 90 degrees flexion). The location of the axis was close to the tibial insertion of the anterior cruciate ligament at 0 degree flexion, gradually moving toward insertion of the posterior cruciate ligament (observed at 45 degrees and 60 degrees flexion), and then moved anteriorly again with additional flexion: the axis was approximately equidistant from the two cruciate insertions at 90 degrees flexion. The results showed that a relatively large degree of tibial rotation was possible in a normal knee and that the location of the axis remained approximately in the area between the two cruciate ligament insertions throughout the range of flexion. However, the location of the axis changed with the flexion angle within this area according to the changes in direction and tension of the cruciate ligaments and the surrounding soft tissues. PMID- 10693566 TI - Augmented amputations of the lower extremity. AB - Ten patients who had amputations of a lower extremity for high-grade sarcomas underwent bone augmentation with either allograft or autograft between 1988 and 1996. There were eight transfemoral amputations and two transtibial amputations. The transferred segments consisted of one proximal tibia and six distal tibia autografts, two allografts, one autograft talar dome and first metatarsal, and one with a patellar cap of a supracondylar amputation. The average length of followup was 54 months. There were no nonunions of any of the grafts. There were three wound problems requiring additional operations. One autograft resorbed, and one autograft had a late infection. There was one local recurrence. Augmentation to provide length resulted in a 42% increase in bone length in those performed purely for length. All patients were able to use standard prostheses. Functional outcome was appropriate to the amputation level. Half of the patients avoided more proximal levels of amputation because of the ability to augment the osteotomy. The use of nonvascularized structural autografts or allografts is a simple procedure that can produce a superior residual limb in patients undergoing amputation. Its use should be considered in patients for whom traditional amputation techniques will result in poor function, difficulty in fitting a prosthesis, or greater than necessary anatomic loss. PMID- 10693567 TI - Fixation of large segment femoral allografts using plates augmented with cerclage wires. AB - Results using plates augmented with cerclage wires to stabilize eight large segment femoral allografts are described. Two patients had plates augmented with cerclage wires placed at the time of tumor resection, and six patients had plates augmented with cerclage fixation placed to manage allograft nonunions or fractures. These internal fixation techniques spanned 10 osteotomy sites: two metaphyseal and eight diaphyseal. The goal of operations that involved internal fixation with plates augmented by cerclage fixation was to obtain osseous union. Osseous union was achieved at both metaphyseal osteotomy sites and at one of the diaphyseal osteotomy sites. Internal fixation of large segment allografts with plates that are augmented with cerclage wires yield poor results when osseous union is the goal of treatment. PMID- 10693568 TI - Cemented hemiarthroplasties for elderly patients with intertrochanteric fractures. AB - The results of 55 consecutive hemiarthroplasties in 54 elderly patients with intertrochanteric fractures were reviewed. The mean age of the patients was 84.2 years (range, 73-99 years). Standard cemented hemiarthroplasty was used and the fractured posteromedial fragment was retained. Two patients were lost to followup and 12 patients died within 6 months of their fractures. The mean length of followup was 13.6 months (range, 6-24.6 months) for the remaining 40 patients (41 hips). Nineteen patients maintained the same walking category as before fracture and eight of these patients had no increase in the dependency on walking aids. The greatest deterioration of walking function occurred in the subgroup of six patients who had a history of confusion and frequent falls. The medical complications are comparable with those described in other series. There were few surgical complications. The authors of this study think that the use of standard cemented hemiarthroplasty is a reasonable alternative to a sliding screw device for the treatment of intertrochanteric fractures. Longer-term studies with larger numbers of patients are required to address the issues of late complications and whether the reconstructions are durable enough for the long-surviving patients. PMID- 10693569 TI - Adverse tissue reactions to bioabsorbable fixation devices. AB - Among 2528 patients operated on using pins, rods, bolts, and screws made of polyglycolic acid or polylactic acid, 108 (4.3%) were affected by a clinically significant local inflammatory, sterile tissue reaction. The three most common indications for the use of these fixation devices were a displaced malleolar fracture, a chevron osteotomy for hallux valgus, and a displaced fracture of the radial head. In 107 patients, the reaction was elicited by a polyglycolic acid implant, and in one patient by a polylactic acid implant. The incidences were 5.3% (107 of 2037) and 0.2% (one of 491), respectively. The adverse tissue responses to polyglycolic acid were seen 11 weeks after the operation, on average, whereas the reaction to polylactic acid occurred 4.3 years after fixation of an ankle fracture. The mild reactions consisted of a painful erythematous papule of a few weeks' duration. Those of medium severity had a sinus that discharged remnants of the implant for up to 6 months. In the patients affected by severe reactions, extensive osteolytic lesions developed at the implant tracks. The histopathologic picture was that of a nonspecific foreign body reaction. In four patients with vigorous reactions, an arthrodesis of the wrist or ankle later was necessary because of severe osteoarthritis. Several markers of increased risk of the occurrence of a foreign body reaction were found. These included a poorly vascularized bone section such as scaphoid, use of a quinone dye as an additive in the polymer, and an implant geometry with large surface area (screw versus pin or rod). For polyglycolic acid implants, the risk of an adverse tissue response in a given clinical situation can be estimated from the findings of this study. For slow degrading polymers like polylactic acid, however, the ultimate biocompatibility still is unsettled, and additional clinical research with long followup is required. PMID- 10693570 TI - Effect of collagen denaturation on the toughness of bone. AB - The purpose of this study was to explore the relationship between the integrity of collagen and biomechanical properties of bone. In this study, age (range, 5-26 years old) and gender related changes in cortical bone samples from 33 baboon femurs (15 males and 18 females) were examined. The percentage of denatured collagen was determined using a selective digestion technique. The fracture toughness, elastic modulus, yield and ultimate strength, and energy to fracture of bone were determined in three-point bending configurations. The porosity and weight fractions of the mineral and organic phase also were measured. A two-way analysis of variance showed that age dependent changes were reflected primarily in the amount of denatured collagen, fracture toughness, energy to fracture, and elastic modulus, whereas gender had effects on the fracture toughness, elastic modulus, and porosity of bone. In addition, regression analyses indicated that the percentage of denatured collagen had an inverse correlation with the toughness of bone and a positive correlation with its elastic modulus, whereas mineral content had positive correlation with the strength and elastic modulus of bone. The results of this study suggest collagen influences the toughness of bone, whereas mineral content predominantly contributes to bone stiffness and strength. PMID- 10693571 TI - Bone graft incorporation. Effects of osteogenic protein-1 and impaction. AB - Impaction of cancellous bone grafts in a bone chamber in rats in a previous study led to decreased ingrowth of new bone after 6 weeks compared with unimpacted grafts. The current study analyzes whether this decrease represented a final loss of ingrowth or just a delay, if the decrease was influenced by immunologic factors, and if it was possible to influence the inhibitory effect by adding a bone morphogenetic protein. Bone chambers with impacted or unimpacted bone grafts were implanted bilaterally in rat tibias. The mean bone ingrowth distance into the graft was measured on histologic sections. Three experiments were done: (1) the bone ingrowth into impacted and unimpacted grafts was studied at 6 and 12 weeks; (2) the immunologic influence was studied by comparing isogeneic grafts with allogeneic grafts; and (3) the authors tried to influence the decrease in bone ingrowth in impacted grafts by adding osteogenic protein-1. Bone ingrowth into the impacted graft was decreased at 6 weeks but not at 12 weeks. No difference was found between isografts and allografts at 6 weeks. With the addition of osteogenic protein-1, the impacted grafts showed dramatically increased bone ingrowth. Impacted bone grafts are incorporated at a slower rate than were structural grafts. The delay can be reversed by adding osteogenic protein-1, making ingrowth faster than in structural bone. PMID- 10693572 TI - Does axial limb rotation affect the alignment measurements in deformed limbs? AB - The long-term outcome of total knee arthroplasty and femoral or tibial osteotomy is related to the ability of the surgeon to achieve the desired alignment based on preoperative planning. This study evaluates the effect of axial rotation on measured tibiofemoral angles and the angle formed between the anatomic and mechanical axes of the femur in lower extremities with valgus and varus deformities. A comparison study of the measured tibiofemoral angles indicated a statistically significant effect in models with severe vagus or varus deformity when rotated 10 degrees internally or externally. In the second part of the study, the measured angle between the anatomic and mechanical axes of the femur never varied by more than 1 degree, despite a 40 degrees are of rotation. The results of the study indicate the tibiofemoral angle measurements are more sensitive to axial limb rotation in lower extremities with valgus or varus deformity than are normally aligned limbs. In preoperative planning of total knee arthroplasty, the measured angle between the anatomic and mechanical axes of the femur is less effected by limb rotation, regardless of the degree of valgus or varus deformity. PMID- 10693573 TI - Buttock pain in a 48-year-old man. PMID- 10693574 TI - Orthopaedics, ethics, and industry. Appropriateness of gifts, grants, and awards. AB - At the Academic Orthopaedic Society meeting in San Francisco on November 8 and 9, 1996, the membership addressed the issue of ethics and industry in an academic setting. Using a Delphi panel technique, they arrived at a definition of conflict of interest, and 41 separate points of acceptable and unacceptable behavior related to gifts, research awards, and funding of various activities. The Academic Orthopaedic Society Delphi Committee also mailed 191 questionnaires (157 department chairpersons and 34 program directors) to 157 training programs. The respective department chairpersons and program directors were asked to copy and distribute the questionnaires to staff (faculty) and house officers (residents and fellows) to complete anonymously and return them for collation. Ninety-one programs (58%) responded. Three hundred and fifty-two questionnaires were returned (237 from staff, 115 from house officers), each of which expressed agreement or lack of agreement with the Delphi panel report using a Likert scale technique. With only modest (and usually predictable) disagreement on certain items, the final statements by the Delphi panel were supported strongly by the survive results. The Academic Orthopaedic Society believes that the major points arrived at by the panelists should serve as the basis for ethical guidelines in the relation between academic orthopaedic institutions and industry. PMID- 10693575 TI - Klein's surgical strike at Medicare. PMID- 10693576 TI - Caveat lector: be wary of media reports about excessive Ritalin use in BC. PMID- 10693577 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693578 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693579 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693580 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693581 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693582 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693583 TI - Putting together the pieces of the physician supply puzzle. PMID- 10693584 TI - Improving management of depression. PMID- 10693585 TI - The move away from fee-for-service care. PMID- 10693586 TI - Assessing quality of care. PMID- 10693587 TI - Prevalence of anemia among James Bay Cree infants of northern Quebec. AB - BACKGROUND: Anemia is common among First Nation infants in Canada, often as a result of iron deficiency, which places them at risk for psychomotor impairment. Prevalence data are unavailable, and the risk factors are unknown. This study assessed the prevalence of anemia and associated risk factors among 9-month-old Cree infants in northern Quebec. METHODS: Between January 1995 and October 1998, 6 of 9 Cree villages in the James Bay region adopted a screening protocol for anemia in 9-month-old infants. Cross-sectional data were obtained from medical charts. The data for babies of very low birth weight and those with fever or infection were excluded. Among the 386 babies whose hemoglobin concentration was known, the type of milk consumed at the time of screening was known for 354. Associations between hemoglobin concentration and mean cell volume at 9 months, and milk type and weight gain since birth were analysed. RESULTS: The mean hemoglobin concentration of the 386 infants was 114.1 (standard deviation [SD] 10.6) g/L. The prevalence of anemia was 31.9% (95% confidence interval [CI] 27.2% 36.7%) with a hemoglobin cutoff value of 110 g/L, 17.6% 95% CI 13.9%-21.7%) with a cutoff value of 105 g/L, and 7.8% (95% CI 5.3%-10.9%) with a cutoff value of 100 g/L. Babies exclusively fed formula at 9 months had a higher mean hemoglobin concentration (118.5 [SD 9.9] g/L) than those exclusively fed breast milk (109.9 [SD 10.0] g/L), cow's milk (112.5 [SD 10.1] g/L) or more than one type of milk (112.0 [SD 10.8] g/L) (p < 0.05). Compared with formula, the odds ratio (OR) for anemia was 7.9 (95% CI 3.4-18.2) for breast milk, 5.0 (95% CI 2.0-12.7) for cow's milk and 5.2 (95% CI 1.9-14.6) for mixed milks. Infants fed formula and those fed cow's milk had significantly greater weight gains since birth, by 724 g and 624 g respectively, than breast-fed infants (p < 0.05). When milk type was controlled for, weight gain since birth was significantly associated with the presence of microcytic erythrocytes (OR comparing highest tertile of weight gain to lowest tertile 2.9, 95% CI 1.2-6.6). INTERPRETATION: Iron-deficiency anemia is highly prevalent among James Bay Cree infants. Measures to increase iron intake are required. PMID- 10693588 TI - Gay and lesbian physicians in training: a qualitative study. AB - BACKGROUND: Gay and lesbian physicians in training face considerable challenges as they become professionalized. Qualitative research is necessary to understand the social and cultural factors that influence their medical training. In this study we explored the significance of gay or lesbian identity on the experiences of medical training using naturalistic methods of inquiry. METHODS: Semi structured interviews, focus groups and an e-mail listserv were used to explore professional and personal issues of importance to 29 gay and lesbian medical students and residents in 4 Canadian cities. Data, time, method and investigator triangulation were used to identify and corroborate emerging themes. The domains explored included career choice, "coming out," becoming a doctor, the environment and career implications. RESULTS: Gay or lesbian medical students and residents experienced significant challenges. For all participants, sexual orientation had an effect on their decisions to enter and remain in medicine. Once in training, the safety of a variety of learning environments was of paramount importance, and it affected subsequent decisions about identity disclosure, residency and career path. Respondents' assessment of professional and personal risk was influenced by the presence of identifiable supports, curricula inclusive of gay and lesbian sexuality and health issues and effective policies censuring discrimination based on sexual orientation. The need for training programs to be proactive in acknowledging and supporting diversity was identified. INTERPRETATION: Considerable energy and emotion are spent by gay and lesbian medical students and residents navigating training programs, which may be, at best, indifferent and, at worst, hostile. PMID- 10693589 TI - Use of physical and chemical restraints in medical teaching units. PMID- 10693590 TI - Breast-feeding and anemia: let's be careful. PMID- 10693591 TI - Education of medical students and house staff to prevent hazardous occupational exposure. PMID- 10693592 TI - Equity in health. PMID- 10693593 TI - Tuberculosis: 13. Control of the disease among aboriginal people in Canada. PMID- 10693594 TI - Safrole in betel quid may be a risk factor for hepatocellular carcinoma: case report. PMID- 10693595 TI - The Internet and evidence-based decision-making: a needed synergy for efficient knowledge management in health care. PMID- 10693596 TI - Septic shock: treating more than just blood pressure. PMID- 10693597 TI - The urea breath test for Helicobacter pylori infection: taking the wind out of the sails of endoscopy. PMID- 10693598 TI - Is 79 too old for a heart transplant? PMID- 10693599 TI - Candy killed after complaints from mentally disabled Americans. PMID- 10693600 TI - Drug costs surpass spending on physicians. PMID- 10693601 TI - Management practices in dental schools: an overview. PMID- 10693602 TI - Patient support services. PMID- 10693603 TI - Health and financial records. PMID- 10693604 TI - Support services and staff responsibilities. PMID- 10693605 TI - Implications and future challenges. PMID- 10693606 TI - Studies of surgical outcome after patellar tendinopathy: clinical significance of methodological deficiencies and guidelines for future studies. Victorian Institute of Sport Tendon Study Group. AB - Patellar tendinopathy is often treated surgically after failure of conservative treatment but clinical experience suggests that results are not uniformly excellent. The aim of this review was to (i) identify the different surgical techniques that have been reported and compare their success rates, and (ii) critically assess the methodology of studies that have reported surgical outcomes. Twenty-three papers and two abstracts were included in the review. Surgical procedures were categorized and outcomes summarized. Using ten criteria, an overall methodology score was derived for each paper. Criteria for which scores were generally low (indicating methodological deficiency) concerned the type of study, subject selection process and outcome measures. We found a negative correlation between papers' reported success rates and overall methodology scores (r= -0.57, P<0.01). There was a positive correlation between year of publication and overall methodology score (r=0.68, P<0.001). We conclude that study methodology may influence reported surgical outcome. We suggest practical guidelines for improving study design in this area of clinical research, as improved study design would provide clinicians with a more rigorous evidence-base for treating patients who have recalcitrant patellar tendinopathy. PMID- 10693607 TI - Effects on the foetus of exercise in pregnancy. AB - Maternal training during pregnancy has been the subject for numerous investigations lately, which are presented in this survey. No studies in human beings have shown any negative effect of training on the embryogenesis. During physical training a small rise in foetal heart rate of 5-25 bpm is a common finding. This could be due to a reduction in oxygen delivery or more likely stimulation from maternal vasoactive hormones or training-induced uterine contractions. Foetal growth seems to be influenced by maternal activity, as some investigations have found significantly bigger babies born by moderately trained females compared to non-trained or heavily trained women. In the latter group the reduction could be explained by a reduced neonatal fat mass. Increased maternal temperature during training has not been found to lead to any foetal abnormalities. The results indicate that moderate training during pregnancy can be recommended with observance of simple directives. PMID- 10693608 TI - Estrogen-dependent tensile properties of the rabbit knee medial collateral ligament. AB - The influence of oophorectomy or continuous administration of estradiol on the tensile properties of the rabbit knee medial collateral ligament was investigated. Young postpubertal female New Zealand white rabbits were either oophorectomized or underwent a sham operation. The sham-operated animals received in addition a daily dosage of 4 mg 17beta-estradiol. After 5 months the animals were killed, and the material properties of the bone-ligament-bone complex in one knee were determined using a material testing machine and video system, and compared to non-treated control animals. There was no difference in elastic modulus between the groups. However, the ligaments from low-dose estrogen-treated animals had a smaller cross-sectional area and a higher ultimate tensile strength than those from controls or oophorectomized rabbits (P<0.04-0.0003). PMID- 10693609 TI - Left ventricular mass, geometry and filling in elite female and male endurance athletes. AB - We compared echocardiographic findings in female (n=30) and male (n=30) endurance athletes to age-matched female (n=15) and male (n=15) sedentary controls. The differences between athletes and controls were similar in both sexes; only left ventricular (LV) mass and septum thickness differed slightly more in men than in women (67% vs 55%, P=0.004, and 36% vs 30%, P=0.03, respectively). LV wall thicknesses were in the normal range in all women, while four (13%) male athletes exceeded 13 mm. In conclusion, the effects of endurance training on echocardiographic findings appear to be quite similar in women and men. However, in female athletes with an abnormally thick left ventricular wall a thorough cardiac evaluation is indicated. This contrasts with male athletes, in whom LV wall thicknesses of over 13 mm are a not uncommon finding. PMID- 10693610 TI - Relationship between perceived readiness to run and physiological variables during repeated 2000 m bouts in middle-distance runners. AB - The aim of this study was to investigate how heart rate (HR) and blood lactate (LA) concentrations are associated with perceived readiness ratings (PRR) to begin a new run during recovery after four different intensity steady-state 2000 m runs in college-level male middle-distance runners (n=15). A typical 4x2000 m run test with stepwise increasing speed was used on the indoor track (150 m lap). A new PRR scale was administered at each minute of recovery. The scale ranges from 1 to 5 points (from "not at all ready to begin" to "completely ready to begin"). Blood LA concentrations were measured immediately after runs and in the 3rd min of recovery after the first and second runs. In case of the third and fourth runs, blood LA was measured immediately after the runs and in the 3rd and 6th min of recovery. HR was recorded at the end of every minute of recovery. Highly significant inverse relationships were revealed between PRR, blood LA concentrations and HR during recovery (r>0.9 as a rule). After the third and the fourth 2000 m runs, where intensity was higher than LA threshold, PRR increased during 6 min up to 4.8+/-0.4 and 4.5+/-0.6, respectively, while HR fell below 120 beats x min(-1). However, blood LA concentration remained high. The reliability of the new PRR scale (tested on four runners), during recovery was very high (r=0.98). These results suggest that the PRR scale can be used by runners to determine the optimal duration of resting intervals between runs. PMID- 10693611 TI - The continuity of physical activity--a retrospective and prospective study among older people. AB - This study investigated the continuity of life-span physical activity by examining the predictors of the maintenance of a high level of physical activity over 8 years among subjects aged 65-84 years at the baseline, in 1988, in Jyvaskyla, Finland. Age, education, marital status and chronic conditions and past physical activity were studied at the baseline. In men and women, self reported competitive sport participation from as early as 10-19 years of age was a significant predictor for maintaining activity in old age. Also women's participation in recreational sports at the age of 40-64 years predicted activity. We concluded that past physical activity is strongly connected to maintaining a high level of physical activity in old age regardless of chronic conditions that may develop. PMID- 10693612 TI - Marathon with cystic fibrosis and bilateral lung transplant. AB - The article presents studies performed before, during and after a marathon run (42,195 m) in a 32-year-old man who underwent a bilateral lung transplantation because of end-stage cystic fibrosis (CF) 15 months prior to the race. Before the run his FEV1 was 81% predicted, compared with 19% predicted before the operation, and his maximal oxygen uptake was 31.9 ml/kg(-1)/min(-1). He completed the New York City Marathon 1998 without major problems in 7 h 8 min 50s. Pulmonary tests, biochemical changes and endocrine responses indicated transient changes, mostly as expected in healthy marathon runners. The case demonstrates that physiological trainability and psychological will power following a successful bilateral lung transplantation can transform a chronically ill CF patient into a robust marathon runner. PMID- 10693613 TI - Comparison between in-line and rollerskating injury. A prospective study. AB - Rollerskating is an activity that has become increasingly popular over the past several years among children and adults in Denmark. During a 7-month period in 1997, 300 in-line skaters and 107 roller skaters were treated in the Emergency Department, Esbjerg County Hospital. Of these, 60.4% had minor injuries (sprains, bruises, lacerations) and 39.6% fractures. There was no statistical significant difference in the types of injury between skater groups. The most common serious injury was fracture of the wrist, which occurred in both skater groups (25%, n=102). Almost all of the fractures of the wrist and elbow occurred among skaters who did not wear wrist or elbow guards Only 20% of the skaters used protective equipment. In-line skaters used protective equipment more often than did roller skaters. Of all accidents, 69% occurred on public roads (street and sidewalk). PMID- 10693614 TI - A comparison of the STAI and CSAI-2 in five-day recalls of precompetition anxiety in collegiate track and field athletes. PMID- 10693615 TI - Topography-controlled excimer laser photorefractive keratectomy. AB - PURPOSE: To assess whether photorefractive keratectomy (PRK) controlled by videokeratography can successfully treat refractive errors in eyes with corneal irregularities and improve spectacle-corrected visual acuity. METHODS: In a prospective clinical study, PRK was performed in 10 eyes of 10 patients. Reason for surgery was irregular astigmatism after penetrating keratoplasty, corneal irregularity after corneal scarring, corneal astigmatism in keratoconus, and decentration after myopic and hyperopic PRK. Excimer ablation was controlled by preoperative videokeratography (Orbscan II, Orbtek) using the MEL-70 system from Aesculap Meditec. Follow-up was 6 months. RESULTS: Concerning manifest refraction, the sphere was reduced on average from +1.92 to +0.57 D, 6 months postoperatively. Cylinder changed from -1.95 D on average to -0.30 D at 6 months postoperatively. There was improvement of uncorrected visual acuity of 2 or more lines in 5 eyes and no change in 5 eyes 6 months postoperatively. Spectacle corrected visual acuity improved in 2 eyes by 2 to 3 lines, in 9 eyes by 1 to 3 lines, and showed no change in 1 eye. CONCLUSION: Videokeratography-controlled PRK improved refractive errors in irregular corneas with improvement of spectacle corrected visual acuity. PMID- 10693616 TI - Corneal lathing using the excimer laser and a computer-controlled positioning system. AB - PURPOSE: To present the excimer laser corneal shaping system (ELCS-S), an add-on device to the Keratom, a commercially available 193-nm excimer laser built by Schwind. METHODS: The system is designed for the preparation of donor corneas under sterile conditions using the ultraviolet laser to offer greatest possible flexibility. Lenticules for planolamellar grafting and refractive epikeratoplasty, as well as donor buttons for penetrating keratoplasty can be computer-designed by the surgeon or technician and lathed with the system. RESULTS: Using the excimer laser corneal shaping system (ELCS-S) on human donor corneas, the central surface of the epikeratoplasty lenticule exhibited only narrow, flat concentric notches corresponding to the single lathing steps. Transmission electron microscopy revealed a damage zone of less than 0.3 microm in close approximation to the treated surface. The final thickness revealed a difference of less than +/-53 microm from the intended, initially programmed value. Ultrastructural studies showed the perpendicular stromal surface of the penetrating keratoplasty buttons to be smooth with minimal protrusion of Descemet's membrane. Endothelial injury was observed in a zone averaging between 40 and 100 microm adjacent to the cutting edge only. CONCLUSION: The excimer laser corneal shaping system (ELCS-S) allows a computer-controlled, surgeon designed, sterile preparation of lamellar and penetrating corneal grafts with the use of the excimer laser. This could offer significant advantages in comparison to presently available systems for lamellar dissection and trephination. PMID- 10693617 TI - Healing after photorefractive keratectomy in cat eyes with a scanning mid infrared Nd:YAG pumped optical parametric oscillator laser. AB - PURPOSE: To evaluate the healing characteristics of cat corneas treated with a new scanning mid-infrared laser system. METHODS: Six adult cats were treated with 6-mm diameter photorefractive keratectomy (PRK) corrections. One eye in each animal was untreated as a control and the other was treated with either a -3.00 or -6.00 diopter ablation. The laser was a new Nd:YAG pumped optical parametric oscillator laser at 2.94 microm with a new scanning delivery system. The pulse width was 7 nanoseconds, the repetition rate was 10 Hz, the size of the laser spot on the eye was 1.0 mm, and the fluence was 150 mJ/cm2. Healing of the cat corneas was followed for 4 months. Slit-lamp and corneal topography evaluations were done at each follow-up examination. Histology was performed at the end of the study. RESULTS: The corneal epithelium healed within 1 week. There was no stromal haze in any eye after the epithelium healed. After the first 2 weeks, slit-lamp examination could not identify which eye was treated. Corneal topography showed corneal flattening. Light microscopy at 4 months revealed normal epithelium and increased keratocyte density in the anterior third of the cornea. Electron microscopy showed discontinuities in the basement membrane and hemidesmosomes. The deep stroma and endothelium were normal. CONCLUSIONS: Cat corneas treated with the new optical parametric oscillator laser healed normally with no adverse effects. Increased keratocyte activity in the anterior stroma was the only noticeable response besides the flattening shown by topography. PMID- 10693618 TI - Histological comparison of corneal ablation with Er:YAG laser, Nd:YAG optical parametric oscillator, and excimer laser. AB - PURPOSE: To use histological techniques to assess and compare the ablation depth, local damage, and surface quality of corneal ablations by a Q-switched Er:YAG laser, an optical parametric oscillator laser at 2.94 microm, a long pulse Er:YAG laser, and a 193-nm excimer laser. METHODS: Human cadaver eyes and in vivo cat eyes were treated with a 6.0-mm diameter, 30-microm-deep phototherapeutic keratectomy ablation and a 6.0-mm diameter, -5.00-D photorefractive keratectomy ablation. Human cadaver eyes were also treated with a 5.0-mm diameter, -5.00-D laser in situ keratomileusis (LASIK) ablation. Fluences and pulse widths used were 200 mJ/cm2 and 70 ns for the Q-switched Er:YAG, 150 mJ/cm2 and 7 ns for the optical parametric oscillator laser (OPO), 500 mJ/cm2 and 50 microseconds for the long pulse Er:YAG, and 160 mj/cm2 and 20 ns for the excimer laser. In the ablation rate study, 12 porcine eyes were ablated by the OPO laser with a range of layers and at different fluences ranging from 60 to 150 mJ/cm2, all using a 1.5-mm spot on the eye. The ablation depth of these acute ablations was evaluated by light microscopy examination. RESULTS: In the acute damage study, light microscopy showed a thin surface layer in all samples with minimal thermal damage except on the long pulse Er:YAG corneas. Transmission electron microscopy revealed less than 0.3-microm surface damage for all specimens of both the optical parametric oscillator and the excimer laser samples with no evidence of collagen shrinkage. Transmission electron microscopy showed damage layers of 0.5 to 3 microm for Q-switched Er:YAG and 3 to 10 microm for long pulse Er:YAG. Scanning electron microscopy showed smooth surfaces in all eyes, although the excimer was the roughest. In the porcine eye study, ablations were produced in both PTK and PRK modes with the ablation rate per layer increasing with the fluence. At 120 mJ/cm2, the average ablation rate was 1.9 microm per layer. CONCLUSIONS: The histology from the short pulse mid-infrared optical parametric oscillator laser at 2.94 microm was comparable to the 193-nm excimer with a smooth, damage-free, ablation zone when performing PRK and LASIK. PMID- 10693619 TI - Limitations of erbium:YAG laser photorefractive keratectomy. AB - PURPOSE: The erbium:YAG laser (lambda = 2.94 microm) has been considered promising as an alternative to the ArF excimer laser in photorefractive keratectomy (PRK). However, corneal application of this mid-infrared solid state laser is still plagued with various disadvantages compared to that of the ArF excimer laser (lambda = 193 nm). We discuss the limitations of PRK with the erbium:YAG laser. METHODS: Measurements of ablation threshold, ablation efficiency, and thermal damage were done to compare the process of erbium:YAG laser photoevaporization to the ArF excimer laser. PRK procedures were performed on fresh enucleated pig corneas to investigate the morphology and surface roughness of the cornea after scanning-spot and fundamental mode photoablation. Surface roughness was measured by using a tactile surface reprofiling system. RESULTS: The ablation threshold and the ablation efficiencies for the erbium:YAG laser are significantly higher compared to the ArF excimer laser. Collateral thermal damage decreases with a reduction of laser pulse duration to a minimum of approximately 5 microm. Scanning electron microscopy and surface roughness measurements of the corneal surface after erbium:YAG laser treatment demonstrated higher surface roughness compared to ArF excimer laser treatments. CONCLUSIONS: The erbium:YAG laser is not at present an alternative to the ArF excimer laser for photorefractive keratectomy. PMID- 10693620 TI - Arcuate transverse keratotomy remains a useful adjunct to correct astigmatism in conjunction with photorefractive keratectomy. AB - PURPOSE: To retrospectively evaluate the effectiveness of paired, arcuate transverse keratotomy (Arc-T) performed prior to or after photorefractive keratectomy (PRK) to correct low to moderate amounts of natural or laser-induced astigmatism. METHODS: Spherical PRK was performed in 730 eyes for myopia of -1.00 to -7.00 D. PRK with arcuate transverse keratotomy was performed in 150 of these eyes; we studied 123 eyes that did not have PRK enhancement. Arc-T was performed prior to PRK in all 37 study eyes with astigmatism of 1.50 D or more at the preoperative examination. Arc-T keratotomy was performed after PRK in 86 study eyes for residual astigmatism of +0.75 D or more and uncorrected visual acuity of 20/30 or worse. RESULTS: Arc-T before PRK group: PRK was performed at a mean 1.0 +/- 1.5 months after Arc-T. Mean astigmatism decreased from +2.40 +/- 0.6 D (range, 1.00 to 4.00 D) before Arc-T to 0.60 +/- 0.60 D (range, 0 to 2.25 D) after Arc-T (P < .0001). Net change in astigmatism was 1.80 +/- 0.60 D (range, 0.80 to 2.80 D) and mean reduction was 75%. Spherical equivalent refraction changed from -4.10 +/- 1.90 D (range, -0.25 to -8.10 D) to -4.40 +/- 1.80 D after Arc-T (P = .002). Mean change in spherical equivalent refraction after Arc-T was 0.30 +/- 0.50 D (range, -1.10 to +0.40 D). Arc-T after PRK group: Arc-T was performed at a mean 3.5 +/- 1.9 months after PRK. Six months after Arc-T, astigmatism was decreased from +1.50 +/- 0.60 D to 0.40 +/- 0.40 D (P = .04). Net change in astigmatism at 6 months was 1.10 +/- 0.60 D and mean reduction was 74%. Vector change in astigmatism magnitude was 1.30 +/- 0.60 D (range, 0 to 4.00 D) at 6 months and vector change in astigmatism axis was 65 degrees +/- 68 degrees. Spherical equivalent refraction did not change when Arc-T was performed after PRK for eyes with low astigmatism (P = .4). Arc-T retreatment was performed in 6 of 37 (16%) eyes that had Arc-T before PRK and 18 of 86 (21%) eyes that had Arc-T after PRK (P = .12). CONCLUSION: Arcuate transverse keratotomy performed prior to PRK for high astigmatism or after PRK for lower levels of residual astigmatism effectively improved visual outcome. Coupling was less predictable for high levels of astigmatism correction with Arc-T. PMID- 10693621 TI - Relative mydriasis after photorefractive keratectomy. AB - PURPOSE: To report the incidence of anisocoria after unilateral excimer laser photorefractive keratectomy (PRK) for myopia and subsequent corticosteroid therapy in a retrospective and prospective study and to explore possible etiologies. METHODS: The horizontal pupil diameter was determined in 6 patients (6 eyes) at 21.8 +/- 12.6 months after unilateral wide-field excimer laser PRK (retrospective group) as well as in 8 consecutive patients (8 eyes) before and 3.4 +/- 2.9 months after unilateral PRK (prospective group). The Schwind-Keratom wide-field excimer laser was used. Measurements were done in an examination room using Rosenbaum card comparison pupillometry and with a Goldmann perimeter at 31.5 asb. In the prospective group, the effect of fitting a hard contact lens of zero diopter power and the application of 0.1% pilocarpine were evaluated. RESULTS: Relative mydriasis was present in all treated eyes and the difference in pupil diameter between the two eyes measured 0.25 to 1.75 mm (retrospective group: +0.56 +/- 0.82 mm; prospective group: +0.72 +/- 0.29 mm). At the time of pupil measurement, the retrospective group had a significantly longer mean postoperative follow-up (21 mo) than the prospective group (3.4 mo) and significantly more eyes still received topical corticosteroid treatment (retrospective group, 1 of 6 eyes; prospective group, 7 of 8 eyes). The amount of anisocoria did not correlate with the applied laser energy, ablation depth, or refractive change, but showed a negative correlation with increasing time after PRK. Neither hard contact lens fitting nor pilocarpine 0.1% reduced the amount of anisocoria significantly. CONCLUSION: Unilateral PRK with wide-field excimer laser ablation and subsequent application of topical corticosteroids regularly resulted in a relative pupillary mydriasis. Neither an altered corneal profile nor parasympathetic denervation is responsible for this. Weakening of the pupillary sphincter of the treated eye may cause this phenomenon. PMID- 10693622 TI - Photorefractive keratectomy for visual rehabilitation of anisometropia induced by retinal detachment surgery. AB - PURPOSE: To evaluate the efficacy of unilateral photorefractive keratectomy to correct anisometropia induced by retinal detachment surgery. METHODS: Photorefractive keratectomy was performed in 10 eyes of 10 patients with anisometropia induced by previous retinal detachment surgery. The Aesculap Meditec MEL 60 excimer laser was used. RESULTS: Preoperative mean spherical equivalent refraction was -5.20 D. Mean postoperative spherical equivalent refraction was -0.25 D after a mean follow-up of 12.9 months. Mean preoperative spherical equivalent refraction difference between two eyes of 4.87 D was decreased to a mean 0.60 D postoperatively (t-test, P < .0001). All patients were free of anisometropic symptoms after laser surgery. CONCLUSION: Unilateral photorefractive keratectomy seems to be an effective method to correct anisometropia induced by conventional retinal detachment surgery, especially for patients with spectacle and contact lens intolerance. PMID- 10693623 TI - Lower intraoperative flap complication rate with the Hansatome microkeratome compared to the Automated Corneal Shaper. AB - PURPOSE: The purpose of this study was to retrospectively compare the incidence of intraoperative flap complications, such as partial flaps, donut-shaped flaps, central corneal cuts, and complete caps with the Hansatome and Automated Corneal Shaper (ACS) microkeratomes. METHODS: All laser in situ keratomileusis (LASIK) procedures performed by a single surgeon with the Hansatome or Automated Corneal Shaper in which intraocular pressure was verified with a pneumotonometer were reviewed. RESULTS: A total of 90 eyes had LASIK with the ACS microkeratome. Six of the ACS eyes (6.7%) had intraoperative flap complications (4 partial flaps, 1 donut-shaped flap, 0 central corneal cuts, 1 complete cap). Partial flaps and donut-shaped flaps were replaced without laser application and the procedure repeated 2 to 3 months later. Two of these eyes lost 2 lines and one lost 1 line of spectacle-corrected visual acuity at 6 months after repeat LASIK. The eye with the donut-shaped flap was treated with transepithelial photorefractive keratectomy (PRK) and had no change in spectacle-corrected visual acuity at 6 months after PRK. The eye with the complete cap had no change in spectacle corrected visual acuity after laser ablation. Five hundred ninety-eight (598) eyes had LASIK with the Hansatome microkeratome. Two of the Hansatome eyes (0.3%) had a flap complication (1 partial flap and 1 donut-shaped flap). The first eye retained spectacle-corrected visual acuity at 6 months after repeat LASIK. The second eye had transepithelial PRK to eliminate the donut shaped flap with no loss of spectacle-corrected visual acuity at 6 months after surgery. The difference in flap complications between the two procedures was statistically significant (P < .01). There were no flap displacements following surgery in either group. CONCLUSION: Intraoperative flap complications are less likely to occur with the Hansatome microkeratome than with the ACS microkeratome. PMID- 10693624 TI - Evaluation of corneal flap dimensions and cut quality using the Automated Corneal Shaper microkeratome. AB - PURPOSE: To evaluate flap dimensions and cut deterioration with repeated blade use in an automated microkeratome. METHODS: The Automated Corneal Shaper (Chiron Adatomed, Munich, Germany), 160-microm plate attached, was used to make a corneal flap in 90 pig cadaver eyes, reusing blades up to five times. Flap diameter was measured by planimetry and thickness was calculated by ultrasound pachymetry. Scanning electron microscopy of stromal beds and blade cutting edges was performed to assess cut deterioration after repeated blade use. RESULTS: Mean flap central thickness was 125 +/- 32 microm. Mean vertical flap diameter was 7.6 +/- 0.4 mm. No correlation was found between thickness and diameter (r = 0.15, P = .45). Progressive thinning of the flap was observed in the direction of the flap hinge. Smooth cuts (using new blades) with periodic chatter lines at the keratectomy edge and in the stromal bed were observed with scanning electron microscopy. Increasing tissue remnants on the stromal bed and decreasing cut quality occurred with repeated blade use. Blades showed larger tissue remnants, nicks, and even folds on the cutting edge proportional to the number of times blades were used. CONCLUSION: Satisfactory cut quality and reproducibility were obtained after a single use of stainless steel blades in the Automated Corneal Shaper microkeratome. Cut quality was degraded dramatically by repeated use of blades. PMID- 10693625 TI - Photospallation: a new theory and mechanism for mid-infrared corneal ablations. AB - PURPOSE: A new mechanism for ablating corneal tissue is proposed, based on photospallation with short pulse mid-infrared (IR) laser radiation. METHODS: By using a judicious combination of high absorption, short pulses, and low fluences, ablation with this process can potentially remove tissue in a highly localized manner with submicron collateral thermal damage characteristics similar to those achieved by excimer lasers. We provide a brief qualitative overview of aspects of the spallation process that distinguish it from the more familiar photoablation and photothermal mechanisms. RESULTS: Results of preliminary parametric analysis based on one-dimensional models of thermoelastic expansion are summarized. CONCLUSION: These preliminary calculations lend support to the conjecture that corneal tissue can be removed effectively with strongly absorbed nanosecond pulses from a mid-IR laser, using operational fluence levels of less than 200 mJ/cm2. PMID- 10693626 TI - Eggink wins 1999 Troutman Award. PMID- 10693627 TI - Central nervous system complications in liver transplant recipients--incidence, timing, and long-term follow-up. AB - BACKGROUND: Neurological impairment is a major source of morbidity and mortality following orthotopic liver transplantation (OLT). We reviewed our experience with neurologic complications among our first 463 consecutive adult OLT recipients. METHODS: Between September 1988 and October 1993, 463 adult patients underwent OLT. Data on incidence, time of onset, and outcome of central nervous system (CNS) complications was obtained from patient charts, including autopsy results when available. CNS complications were classified by clinical presentation and by etiology. RESULTS: 93 patients (20.1%) had CNS complications following OLT. Encephalopathy (11.8%) and seizure (8.2%) were the leading complications. The incidence of immunosuppressive drug-related complications was 5.6%; coma, 1.7%; cerebral hemorrhage, 1.5%; central pontine myelinolysis (CPM), 1.2%; stroke, 0.6%; and primary CNS lymphoma, 0.2%. Most CNS events (80%) were encountered in the first month after OLT. In the majority of cases, encephalopathy (70%) and seizure (50%) presented in the first 2 wk. Although most CNS infections occurred early, 2 patients developed tuberculous meningitis more than 1 yr post-OLT. In 12 patients, death was directly related to CNS complications (2.6%). CONCLUSIONS: Most CNS complications occur early following OLT but may be seen even after 1 yr. Patients may survive serious neurologic events, such as cerebral hemorrhage, CPM, and meningitis. PMID- 10693628 TI - Pulmonary Rhizopus infection in a diabetic renal transplant recipient. AB - Infectious complications after renal transplantation remain a major cause of morbidity and mortality. Mucormycosis is a rare infection in renal transplant recipients; however, mortality is exceedingly high. Risk factors predisposing to this disease include prolonged neutropenia, diabetes, and patients who are immunosuppressed (Singh N, Gayowski T, Singh J, Yu LV. Invasive gastrointestinal zygomycosis in a liver transplant recipient: case report and review of zygomycosis in solid-organ transplant recipients, Clin Infect Dis 1995: 20: 617). Life-threatening infections can occur, as this fungus has the propensity to invade blood vessel endothelium, resulting in hematological dissemination. We report a case of cavitary Rhizopus lung infection, 2 months after renal transplantation, where the patient was treated successfully with Amphotericin B and surgical resection of the lesions with preservation of his allograft function. In this era of intensified immunosuppression, we may see an increased incidence of mucormycosis in transplant population. Invasive diagnostic work-up is mandatory in case of suspicion; Amphotericin B and, in selected cases, surgical resection are the mainstays of therapy. PMID- 10693629 TI - Iliac artery stenosis masquerading as diuretic resistant congestive heart failure. AB - Iliac artery stenosis is a rare cause of renal dysfunction in renal allograft recipients. Its presence can mimic renovascular hypertension and yet alter the very radiologic tests used to diagnose renal artery stenosis. We investigate a case of iliac artery stenosis that presented with diuretic resistant fluid overload, hypertension and limb claudication that exposed the pitfalls in the diagnosis and management of this condition. Successful stent placement, 8 months after transplant, resulted in return of the serum creatinine below the post transplant nadir. PMID- 10693630 TI - Influence of anti-rejection therapy on the timing of cytomegalovirus disease and other infections in renal transplant recipients. AB - Infections are an important cause of mortality and morbidity in renal transplant recipients. To study the impact of anti-rejection therapy on the timing of infections, the records of 599 consecutive renal transplants, performed prior to 31 December 1996 at the Royal Melbourne Hospital, were reviewed. Patients were grouped according to acute rejection (AR) episode and treatment during the first 6 months after transplantation. Group 1 [n = 168 (35%)] patients did not experience any episode of AR. Group 2 [n = 169 (35%)] patients had one or more episodes of AR and received high doses of steroids. Group 3 [n = 141 (30%)] patients had more than one episode of AR and received anti-lymphocyte antibodies in addition to high doses of steroids. Infections were more common in Groups 2 and 3 but only cytomegalovirus (CMV) disease occurred earlier in patients treated with lympholytics. Given the high incidence and early onset of CMV disease in patients receiving lympholytics and considering that an effective prophylactic protocol remains undetermined, pre-emptive treatment with ganciclovir in this high risk group appears justified. PMID- 10693632 TI - Commerce in transplantation: how does it affect European legislation? AB - Commerce in transplantation is well known, if not well defined. Although the word commerce suggests an exchange of money, in reality it often simply signifies a non-profit-making transaction. Nevertheless, money, and therefore profit, may be involved in some human organ transactions, and the buying and selling of organs for transplantation remains common in too many countries. Clearly, if such transactions were allowed to continue only those who could afford to pay would benefit. They would probably also lead to an increase in the number of media horror stories. A number of such stories have appeared in the past. Although they are rarely based on hard evidence, they do influence politicians and, as a consequence, affect legislation and the availability of organs for transplant. They may also diminish the willingness of the general public to become organ donors and contribute to the persistent poor supply of organ donors. Organ exchange organizations, such as Eurotransplant, have made many efforts to prevent unethical transactions. Nevertheless, stories of such transactions continue to appear and are unlikely to abate while there is a high demand and poor supply of organs for transplantation. An international donor surveillance committee--a clearing house for information on malpractice--could be one solution to the problem as it would prevent doctors from taking part in unethical transplant procedures. PMID- 10693631 TI - Cytomegalovirus immune globulin after liver transplantation: a cost-effectiveness analysis. AB - OBJECTIVE: Cytomegalovirus (CMV) immune globulin (CMVIG) has been shown to significantly reduce severe CMV-associated disease complicating orthotopic liver transplant (OLT). We evaluated the economic impact of severe CMV-associated disease and calculated the marginal cost-effectiveness (C/E) of routine prophylaxis with CMVIG after OLT. DESIGN: C/E analysis. SETTING: Four teaching hospitals in Boston. PATIENTS: Patients who underwent OLT from January 1988 through June 1990. MEASUREMENTS: We gathered actual cost data of hospital care for patients enrolled in a clinical trial of CMVIG prophylaxis in OLT. We calculated average outpatient expenses from a separate group of patients undergoing OLT and developed a regression model to estimate costs during the first year post-transplant (R2 = 0.77). Based on this model, we calculated variable costs (in 1999 US dollars) for all patients in the randomized trial. From the published literature we obtained the probability of CMV outcomes and of long-term survival after OLT. We then developed a decision analytical model to determine an incremental C/E ratio, using a Markov simulation to estimate long term survival and long-term costs. We discounted costs and life-years at 3% and 5% per yr. RESULTS: Based on the efficacy rate of 54% in the controlled trial, we estimate that CMVIG will increase life expectancy by 0.65 discounted years at an additional cost of $11,600, providing a marginal C/E ratio of $17,900/yr life saved. Examining the confidence limits of efficacy, we estimate that CMVIG will have a marginal C/E ratio of $66,200 gained/yr at an efficacy of 11% and $14,000 gained/yr at an efficacy of 83%. CONCLUSION: After OLT, prophylactic CMVIG has an incremental C/E ratio comparable to that of other well-accepted medical therapies and should be used routinely in these patients. PMID- 10693633 TI - Delphi-panel analysis of appropriateness of high-dose chemotherapy and blood cell or bone marrow autotransplants in women with breast cancer. AB - BACKGROUND: There is controversy whether high-dose chemotherapy and a blood cell or bone marrow autotransplant is a better treatment than conventional-dose chemotherapy for women with local/regional or metastatic breast cancer. Subject selection and time-to-treatment biases make definitive comparison impossible. Recent results of randomized trials are contradictory. OBJECTIVE: Determine appropriateness of high-dose chemotherapy and a blood cell or bone marrow autotransplant in women with breast cancer. PANELISTS: Nine breast cancer experts from diverse geographic sites and practice settings. EVIDENCE: Boolean MEDLINE searches of 'breast cancer' and 'chemotherapy' and/or 'blood cell' or 'bone marrow transplants'. PROCESS: We used a modified Delphi-panel group judgement process. Clinical variables were permuted to define 2058 clinical settings. Each panelist rated appropriateness of high-dose therapy and an autotransplant versus conventional therapy on a 9-point ordinal scale (1: most inappropriate, 9: most appropriate). An appropriateness index was developed based on median rating and amount of disagreement. The relationship of appropriateness indices to the permuted clinical variables was considered by analysis of variance and recursive partitioning. CONCLUSIONS: In women with local/regional breast cancer autotransplants were rated: 1) appropriate in those with > or = 10 cancer involved lymph nodes; 2) uncertain in those with 4-9 cancer-involved nodes; and 3) inappropriate in women with < or = 3 cancer-involved lymph nodes. In women with metastatic breast cancer autotransplants were rated: 1) appropriate in those with metastases to 'favorable' sites (skin, lymph node, pleura) and a complete or partial response to chemotherapy; 2) uncertain in women with metastases to 'unfavorable' sites (lung, liver, or central nervous system) and a complete response to chemotherapy or those with bone metastases and a complete or partial response or stable disease after chemotherapy; and 3) inappropriate in other settings. PMID- 10693634 TI - Preliminary experience with midodrine in kidney/pancreas transplant patients with orthostatic hypotension. AB - In an effort to ameliorate the problem of orthostatic hypotension in pancreas transplant patients, current medical management consists of maximizing the patient's hydration, altering antihypertensives, increasing sodium intake, initiation of fludrocortisone, compression stockings, and behavioral modifications. Despite these medical interventions, a subset of patients remains symptomatic. Midodrine (ProAmatine), an alpha-adrenergic agonist, was approved for the treatment of symptomatic orthostatic hypotension in the US. This preliminary report attempts to assess the safety and efficacy of midodrine use in kidney/pancreas (KP) or pancreas alone (PA) transplant recipients. A retrospective review was performed of 7 KP and 1 PA recipient experiencing symptomatic postural hypotension after maximizing other medical treatments. Blood pressure, serum creatinine (SrCr), and objective responses to postural hypotension were assessed at routine intervals. Pre-midodrine monitoring revealed a mean orthostatic change in systolic blood pressure from sitting to standing of 43 mmHg (range 20-100 mmHg). Patients received a mean starting midodrine dose of 18 mg/d, which was titrated to a maximum dose of 30 mg/d. Systolic blood pressure monitoring revealed a mean orthostatic change of 27 mmHg (range 0-81 mmHg) after initiation of treatment with midodrine and a mean follow-up of 3.2 months. All study patients reported improvement in symptoms of orthostatic hypotension. SrCr was not affected based upon comparison of pre-treatment and current SrCr values of 1.4 and 1.3 mg/dL, respectively. The most common side effect experienced was supine hypertension. These preliminary results suggest that midodrine is safe and effective in transplant recipients; however, the dosage should be titrated to symptomatic relief or a maximum dose of 30 mg. Careful monitoring for supine hypertension is necessary. PMID- 10693635 TI - Quality of life in long-term survivors after liver transplantation: impact of recurrent viral hepatitis C virus hepatitis. AB - Post liver transplant recurrence of infection with hepatitis C virus (HCV) occurs in approximately 50% of patients transplanted because of HCV-related liver disease. The aim of this study was to assess long-term quality of life, psychologic distress, and coping in patients with recurrent HCV after liver transplantation in comparison to patients transplanted for other etiologies of underlying liver disease. All liver transplant recipients transplanted at a University affiliated Veterans Affairs Medical Center who had greater than 6 months follow-up were sent a questionnaire investigating quality of life (assessed by Medical Outcomes study health survey SF-36), depression (assessed by Beck Depression Inventory), total mood disturbance (assessed by Profile of Mood States scale), coping (assessed by Billing and Moos Inventory of coping with illnesses), and employment status. Lower Beck Depression Inventory score (p = 0.001), lower mood disturbance score (p = 0.0001), overall satisfaction with present work (p = 0.0001), and lesser use of avoidant coping (p = 0.06) were predictors of better quality of life in long-term survivors of liver transplantation. At a mean follow-up of 4 yr after liver transplantation, patients with histopathologically diagnosed recurrent viral HCV hepatitis had significantly lower global quality of life score (mean score of 76.4 versus 86.2, p = 0.011) and physical functioning score (mean score 20 versus 25, p = 0.015), as compared to all other patients. In summary, quality of life and physical functioning were significantly impaired in liver transplant recipients with histopathologically diagnosed recurrent HCV hepatitis, as compared to those whose HCV hepatitis had not recurred or those transplanted for other reasons. PMID- 10693636 TI - Infusion of donor spleen cells and rejection in liver transplant recipients. AB - Intact or inactivated donor lymphoid cells have been found to downregulate the alloimmune response in a number of experimental models. We conducted a randomized, prospective, double blind, and placebo-controlled trial to determine whether heat-treated donor spleen cells would affect early rejection after liver transplantation. Donor spleen was obtained during organ procurement for 40 patients undergoing liver transplantation. All patients were treated with cyclosporine, azathioprine and steroids. The patients were randomized after surgery to receive either heat-treated (45 degrees C for 1 h) spleen cells or placebo. Patients underwent protocol biopsies at 1 wk, 4 and 12 months, or as needed. Biopsies were reviewed in a blind fashion and scored according to the Banff consensus criteria. Randomization resulted in 19 patients in the spleen cell group and 21 in the placebo group. One-yr graft survival was 94 and 100%, respectively. Early rejection was more frequent in the spleen cell group (61 vs. 35%, p, not significant). The histopathological rejection activity index at 7 d was also higher for the patients in the spleen cell group: 39% of spleen cell treated patients had a score of 4 or higher as opposed to 5% in the placebo group (p < 0.01). The mean score was 2.9 +/- 2.8 for the spleen cell group versus 1.3 + 1.7 for the placebo group (p = 0.034). It is concluded that heat-treated donor spleen cells given within 24 h after liver transplantation were not clinically beneficial and increased the intensity of rejection in 7-d protocol liver biopsies. PMID- 10693637 TI - Immunization of renal transplant recipients with pneumococcal polysaccharide vaccine. AB - BACKGROUND: Streptococcus pneumoniae, a common pathogen leading to pneumonia, is a cause of morbidity and mortality in immunosuppressed patients. Vaccination against this agent can be recommended for immunosuppressed patients, including those with chronic renal failure, nephrotic syndrome and renal transplant recipients; however, a diminished immune response and loss of protective antibodies have been observed. PATIENTS AND METHODS: In our prospective study, the efficacy and side effects of polyvalent pneumococcal vaccination were investigated in renal transplant recipients. A total of 21 patients (6 female, 15 male) with well-functioning renal allografts, who had transplant surgery at least 2 months before, were included in the study. The patients were stratified according to the immunosuppressive protocol and 8 received double, while 13 received triple, immunosuppressive agents. After obtaining basal serum samples, all cases were vaccinated with the 0.5 mL intramuscular administration of polyvalent polysaccharide pneumococcal vaccine (Pneumo 23 Pasteur Merieux, lot No: K 1131). RESULTS: Following a mean of 6 wk in all patients and also a mean of 12 wk in 12 patients, serum samples were again obtained to measure pneumococcal antibodies. Antibody titers following 6 and 12 wk of vaccination were significantly higher, as compared with basal values in all patients, except one. These titers did not show any statistically significant difference between double and triple therapies. There was no significant difference between the 12th and 6th wk postvaccination antibody titers. No systemic or local adverse effects were observed. CONCLUSION: Pneumococcal vaccination is safe and effective in patients with well-functioning renal allografts, at least in the short term. This vaccination policy may be useful for preventing invasive pneumococcal disease in immunosuppressed patients. PMID- 10693638 TI - A longitudinal study of TGF-beta1 protein levels in renal allograft recipients converted from CsA to MMF or AZA. AB - Cyclosporine (CsA) is thought to enhance transforming growth factor (TGF)-beta1 production in vitro and in vivo and this may have a negative effect on long-term graft survival. Therefore, we studied TGF-beta1, plasma levels in 30 patients before kidney transplantation, after transplantation during CsA treatment and after conversion from CsA to azathioprine (AZA) or mycophenolate mofetil (MMF). We questioned whether TGF-beta1 plasma levels would decrease after the discontinuation of CsA and whether the TGF-beta1 plasma levels did correlate with CsA trough levels and kidney function, measured by serum creatinine levels. TGF beta1 plasma levels measured 1 yr after transplantation were lower compared to levels measured before transplantation, however not significantly (p = 0.08). After conversion from CsA to MMF or AZA, a slight increase was observed in some patients, but in the total group TGF-beta1 levels remained unaffected. No correlation was found between the TGF-beta1 levels and CsA trough levels nor with creatinine levels. In conclusion, we did not observe higher TGF-beta1 plasma levels in plasma levels of patients receiving CsA treatment compared to blood from the same patients while on AZA or MMF. PMID- 10693639 TI - Administration of prostacyclin after liver transplantation: a placebo controlled randomized trial. AB - The shortage of suitable organs for liver grafts is responsible for the use of marginal donors for liver transplantation (OLT). If these liver grafts function poorly initially after OLT, a supportive therapy is necessary. The purpose of this study was to evaluate the effects of prostacyclin (PGI2) on postoperative liver graft function after OLT. A total of 30 adult recipients of primary OLT were randomized to either receive PGI2 (4 ng/kg per min body weight, n = 15) or a placebo for 6 d. To evaluate regional splanchnic oxygenation a fiberoptic pulmonary-artery catheter was inserted into a hepatic vein and the difference between mixed venous oxygen content and hepatic venous oxygen content was determined (deltaO2). Measurements were performed directly after transplantation and at 6, 12, 24 and 48 h postoperatively. A significant correlation between deltaO2 and the level of transaminases (ALT/AST) was observed 24 and 48 h after transplantation (p < 0.05). PGI2 treatment induced a significant decrease in deltaO2 after 24 and 48 h after reperfusion (p < 0.05). Peak AST levels tended to be lower in the PGI2 treatment group (418 +/- 99 vs. 638 +/- 156 U/L, p < 0.1). These results suggest that administration of PGI2 after OLT improves hepatic splanchnic oxygenation and may thereby reduce reperfusion injury after OLT. PMID- 10693640 TI - Steroid withdrawal in pancreas transplant recipients. AB - BACKGROUND: Numerous studies of steroid withdrawal have been carried out in kidney and liver transplant recipients, but only a few in pancreas transplant recipients. Yet, pancreas transplant recipients could have significant long-term benefits from steroid withdrawal. METHODS: We performed a retrospective analysis to determine the feasibility of steroid withdrawal in pancreas transplant recipients. RESULTS: Of 360 recipients who underwent a pancreas transplant between January 1, 1994 and June 30, 1998, 14 attempted steroid withdrawal (12 simultaneous pancreas-kidney [SPK]; 2 pancreas transplant alone [PTA]). Reasons for steroid withdrawal were bone fractures (n = 3), psychiatric disorders (n = 2), severe acne (n = 1), recurrent infections (n = 4), and problems with hypercholesterolemia or hypertension (n = 4). All 14 were maintained on tacrolimus and mycophenolate mofetil (MMF) immunosuppression, except for 1 who was on tacrolimus and azathioprine (AZA). Of the 14 recipients, 11 had no episodes of acute rejection before steroid withdrawal. The remaining 3 had one or more acute rejection episodes. Of the 14 recipients, 10 (72%) currently remain off steroids (mean follow-up 18 months, range 5-51 months). However, 4 recipients have resumed steroids: 2 after an acute rejection episode (at 2 and 21 months post-withdrawal) and 2 because of leukopenia (WBC < 3000) and an inability to tolerate full-dose MMF. Steroid withdrawal was unsuccessful in both PTA recipients and in 2 of the 12 SPK recipients. All 14 recipients currently have a functioning pancreas graft. However, 1 of the SPK recipients, in whom steroid withdrawal failed, has developed chronic kidney rejection and is now back on hemodialysis awaiting a retransplant. CONCLUSION: Steroid withdrawal is possible in up to 70% of pancreas transplant recipients. Further studies are necessary to define ideal candidates for steroid withdrawal. Based on the results of this analysis, we have launched a prospective, randomized trial of steroid withdrawal in pancreas transplant recipients. PMID- 10693641 TI - Successful cadaveric renal transplantation of patients highly sensitized to HLA Class I antigens. AB - The purpose of our investigation was to evaluate long-term graft survival and the role of histocompatibility in patients who were highly sensitized to human leukocyte antigen (HLA) Class I antigens and received a cadaveric renal transplant. Our multi-institutional study evaluated 7-yr graft outcomes and the histocompatibility requirements of 61 (6.1%) highly sensitized (anti-human globulin panel reactive antibody [AHG PRA], > or = 80%) cadaveric renal transplantation patients, transplanted between 1988 and 1997, among 999 consecutive cadaveric renal transplants. One- and 7-yr graft survival in the high PRA group (n = 61) was 76 and 59%, and was not significantly different from that in the low PRA group (n = 938), 86 and 59% (Wilcoxon = 0.11; log-rank = 0.45) (died with a functioning graft [DWFG] not censored). When those data were divided into primary and regrafts, 1- and 7-yr graft outcomes for high and low PRA groups were not significantly different [(primary, 1- and 7-yr survival: high PRA = 83 and 74%, n = 30, and low PRA = 87 and 61%, n = 825; log-rank = 0.37 for DWFG not censored) (regrafts, 1- and 7-yr survival: high PRA = 70 and 42%, n = 31, and low PRA = 80 and 43%, n = 113; log-rank = 0.36 for DWFG not censored)]. We did observe a subgroup of the high PRA patient group that had inferior graft outcomes. Graft outcome at 1 and 6 yr in the high PRA group for patients who had one to two DR mismatches (65 and 50%, n = 41) was significantly worse than for high PRA patients who had zero DR mismatches with their donors (100 and 78%, n = 20) (log-rank = 0.01 for DWFG not censored). Furthermore, the mean number of HLA A and -B mismatches was significantly greater in the high PRA/DR-mismatched group (1.7 +/- 1.2, n = 41) compared with the high PRA/zero DR-mismatched group (0.5 +/ 1.1, n = 19) (p < 0.001). Overall, these data suggest that the patient who is highly sensitized to HLA Class I antigens has a long-term graft outcome that is equivalent to less sensitized patients, but that HLA-DR mismatching and a higher degree of Class I mismatching may be poor prognostic indicators in such patients. PMID- 10693642 TI - Monitoring of anti-HLA class I and II antibodies by flow cytometry in patients after first cadaveric kidney transplantation. AB - While the relevance of pre-formed anti-human leukocyte antigen (HLA) antibodies has been studied extensively, the role of anti-HLA class I and II antibodies produced after cadaveric kidney transplantation is still a matter of discussion. As it has been proposed that they are involved in a considerable number of cases, it should be investigated whether a post-transplant monitoring is a sensitive parameter for the early diagnosis of acute rejection episodes. Additionally, it has been suggested that antibodies are a major cause for chronic rejection; thus, it would be of interest to correlate antibody detection and graft survival. We retrospectively investigated 59 patients after a first cadaveric kidney transplantation without known anti-HLA antibodies (complement-dependent cytotoxicity [CDC] testing). The panel reactivity was determined with a new highly sensitive and specific flow-cytometric technique (Flow-PRA Screening Test, One Lambda, Canoga Park, USA) in sequentially collected serum samples pre- and post-transplant. In patients with acute rejection episodes during the clinical course, the last sample prior to rejection, and in patients without rejection, the last sample prior to discharge, was analyzed. Furthermore, we analyzed 3-yr graft survival and several clinical parameters such as cold ischemia time (CIT). Twenty-four of 59 patients (41%) experienced acute rejections during the clinical course. Five of 59 died with a functioning graft within the first 3 yr. Seven of 54 patients, still alive after 3 yr, lost their graft. Anti-HLA antibodies were detectable in only 7/59 patients and a correlation between antibody positivity and acute rejections (p = 0.32 and 0.54 for anti-HLA class I and II, respectively) could not be identified (sensitivity 12.5 and 8.3%). However, we found a significant correlation between the detection of anti-HLA class II and graft loss within 3 yr (p = 0.005, specificity 97.9%). Additionally, anti-HLA class II positive patients had significantly longer CIT (p = 0.003). Whether the detection of anti-HLA class II antibodies in the early post-transplant phase is of great value for the identification of patients at high risk for early graft loss needs additional investigation. However, we found that anti-HLA antibodies are detectable only in a minority of unsensitized patients and we conclude that flow-cytometric monitoring with Flow PRA is not a sensitive parameter for the early diagnosis of acute rejection episodes in patients after first cadaveric kidney transplantation. PMID- 10693643 TI - Should I accept this kidney? AB - BACKGROUND: Transplant candidates frequently ask whether they should, based on information available at the time, accept a cadaver kidney or wait for a potentially better one. METHODS: We analyzed 937 first and second cadaver transplants done between January 1, 1984 and December 31, 1997 to determine if information available at the time an offer is made could be used to predict long term graft survival. RESULTS: By Cox regression, risk factors for worse long-term graft survival were older donor age, cardiovascular or cerebrovascular cause of donor death, and delayed graft function (DGF). HLA-ABDR mismatch was marginally significant. Whether DGF will occur is not known at the time of an offer, but risk factors can be determined; we found these to be older donor age and > 10% panel-reactive antibodies (PRA) at transplantation (by Cox regression). Using these variables (PRA, ABDR mismatch, donor age, and donor cause of death) known at the time of an offer, we calculated the relative risk of worse long-term graft survival for each subgroup (Table 3 in manuscript). In general, older age and donor death from cardiovascular or cerebrovascular disease were associated with worse outcome. Kidneys from donors of < 50 yr had the best outcome, irrespective of match. CONCLUSION: The data provided can be used to help guide patients as to whether they are better off accepting an offered kidney or waiting for a potentially better one. If an offer is declined, the next kidney may have a potentially worse outcome. PMID- 10693644 TI - The role of family and friend social support in reducing risk behaviors among HIV positive Gay men. AB - Researchers have intimated a link between social support and risk-taking behaviors for HIV-positive persons, yet few have empirically examined this assumption. This study examined HIV-positive Gay men regarding their perceptions of family and friend social support, behavioral intentions, and risk-taking behaviors. Results indicated that the more family members were perceived as supportive the less likely participants intended to behave in risky ways. In addition, family availability for support was more predictive of reduced risky behaviors than the availability of friends. Implications for researchers and helping professionals are discussed. PMID- 10693645 TI - Sexual risk behaviors of Gay, Lesbian, and bisexual youths in New York City: prevalence and correlates. AB - The lifetime and recent sexual risk behaviors of 156 Gay, Lesbian, and bisexual youths, recruited from Gay-focused organizations in New York City, were examined. The data indicated seven reasons why the youths are at risk for HIV and other STDs: They initiated sex during early adolescence; their first sexual partners were older than they were; HIV barrier methods (e.g., condoms) were initiated subsequent to sexual debut; many lifetime sexual partners and encounters were reported; some youths exchanged sex for goods; many youths reported having had at least one partner at risk for HIV; and the youths engaged in unprotected sexual behaviors during the past 3 months. Significant gender differences emerged (e.g., the male youths reported more lifetime same-sex partners than the female youths; the female youths reported more lifetime other-sex partners than the male youths). Recent sexual risk behaviors (i.e., numbers of same-sex partners, encounters, and unprotected sex during the past 3 months) were related significantly to the youths' average degree of emotional involvement in or average duration of intimate relationships. PMID- 10693646 TI - Developing, implementing, and evaluating a condom promotion program targeting sexually active adolescents. AB - This article describes the development, implementation, and evaluation of the Condom Campaign, a 1995 HIV prevention program promoting condom use among sexually active adolescents in three King County, Washington, urban communities. This program employed three main strategies: (a) mobilizing all levels of the target communities to support and guide program development and implementation; (b) creating and implementing a mass media campaign targeting sexually active teenagers that promoted correct condom use and favorable attitudes toward condoms; and (c) recruiting public agencies, community organizations, and businesses to distribute condoms from bins and vending machines. We evaluated the program through a series of cross-sectional interviews conducted in the three communities chosen for their elevated levels of adolescent sexual risk behavior. Overall, 73% of target youth reported exposure to the Condom Campaign; exposure did not differ by age, gender, race, or level of sexual experience. Levels of sexual activity remained stable throughout the media campaign. PMID- 10693647 TI - What high-risk women are telling us about access to primary and reproductive health care and HIV prevention services. AB - Focus group discussions on barriers to health care and attitudes toward family planning, reproductive health services, and condom use were conducted with 63 women at high risk for HIV due to their own injection drug use, sex with injection drug users, sex industry work, or a history of multiple sexually transmitted diseases. Barriers identified include the high cost of health care, perceived poor quality of care and experiences of discrimination and stigmatization, geographic accessibility, fear of legal/social services punitive actions, misperceptions about the efficacy of birth control methods and condom usage, lack of sterilization services, and lack of male involvement. Where possible, findings from the focus groups are supported with quantitative survey data from a sample of high-risk women (n = 723). Recommendations are made for improving care for high-risk women. PMID- 10693648 TI - The AIDS Memorial Quilt as preventative education: a developmental analysis of the Quilt. AB - This study consisted of a survey given to college students (N = 560) at a rural university in the Pacific Northwest. The sample was randomly assigned into four groups, following the Solomon four-group study design. The two levels of treatment included interventions consisting of a visit to the AIDS Memorial Quilt for the experimental groups and attendance at an unrelated event for the control groups. Pretests were completed 4 weeks prior to interventions; posttests were completed by the entire sample 4 weeks after the interventions. Results confirmed expected differences among the four groups in terms of social distance, perceptions of people with AIDS, self-efficacy, and discussion of risky behavior. The results suggest that the AIDS Memorial Quilt addresses issues centrally related to behavior change and indicates support for the message interpretation process and stages of change models. PMID- 10693649 TI - Oral health perceptions and adherence with dental treatment referrals among caregivers of children with HIV. AB - Results from a 3-year longitudinal study on the oral manifestations of AIDS (OMA) among seropositive children and their siblings indicated poor adherence with recommendations for dental treatment (Broder, Catalanotto, Reisine, & Variagiannis, 1996). The purposes of this study were to (a) to examine oral health behaviors, attitudes, and perceived barriers to care among caregivers of children with HIV and their siblings who were referred for dental care, and (b) develop and evaluate a 5-week summer pilot program to increase adherence with referral for dental treatment. Telephone interviews with caregivers were conducted to identify barriers to care and to implement services to increase attendance in the dental clinic for their children. Interviews were completed with 28 of the 38 (74%) caregivers recruited from the OMA study (previously cited) who had children referred for dental treatment at the final (sixth) oral health research exam. Twelve of their 58 children (21%) had obtained dental care privately, 25 (62.5%) initiated treatment and 2 (6.3%) completed treatment at the referred dental school during the 5-week pilot program. Although caregivers of children with HIV and their siblings were responsive to the initial efforts of the program's service coordinators, follow-up data from the coordinators' records and chart abstraction revealed that the majority of the participants did not appear for their second or third appointments. The interview reports suggested that caregivers expect dental treatment, such as restorations, at each appointment and do not regard exams/treatment planning as treatment. Personal/family and health care delivery system factors were expressed barriers to dental care. Implications for future programs and investigations are discussed. PMID- 10693650 TI - The herpesvirus proteases as targets for antiviral chemotherapy. AB - Viruses of the family Herpesviridae are responsible for a diverse set of human diseases. The available treatments are largely ineffective, with the exception of a few drugs for treatment of herpes simplex virus (HSV) infections. For several members of this DNA virus family, advances have been made recently in the biochemistry and structural biology of the essential viral protease, revealing common features that may be possible to exploit in the development of a new class of anti-herpesvirus agents. The herpesvirus proteases have been identified as belonging to a unique class of serine protease, with a Ser-His-His catalytic triad. A new, single domain protein fold has been determined by X-ray crystallography for the proteases of at least three different herpesviruses. Also unique for serine proteases, dimerization has been shown to be required for activity of the cytomegalovirus and HSV proteases. The dimerization requirement seriously impacts methods needed for productive, functional analysis and inhibitor discovery. The conserved functional and catalytic properties of the herpesvirus proteases lead to common considerations for this group of proteases in the early phases of inhibitor discovery. In general, classical serine protease inhibitors that react with active site residues do not readily inactivate the herpesvirus proteases. There has been progress however, with activated carbonyls that exploit the selective nucleophilicity of the active site serine. In addition, screening of chemical libraries has yielded novel structures as starting points for drug development. Recent crystal structures of the herpesvirus proteases now allow more direct interpretation of ligand structure activity relationships. This review first describes basic functional aspects of herpesvirus protease biology and enzymology. Then we discuss inhibitors identified to date and the prospects for their future development. PMID- 10693651 TI - Plant-derived and semi-synthetic calanolide compounds with in vitro activity against both human immunodeficiency virus type 1 and human cytomegalovirus. AB - Plant-derived and semi-synthetic calanolide compounds with anti-human immunodeficiency virus type 1 (HIV-1) activity were tested for anti-human cytomegalovirus (HCMV) activity in both cytopathic effect inhibition and plaque reduction assays. The results indicated that the anti-HCMV activity of calanolide compounds does not correlate with their activity against HIV-1. The semi synthetic 12-keto derivatives tended to be more active against HCMV than the corresponding 12-OH congeners, which were more active against HIV-1. It appeared that the 7,8-unsaturated double bond in the chromene ring played a certain role in maintaining activities against both HCMV and HIV-1. Saturation of the double bond increased the EC50 values against both viruses, with concomitant increase in toxicity. The calanolide compounds reported here are the first non-nucleoside analogues capable of inhibiting both HIV-1 and HCMV and, therefore, may be useful chemoprophylactic agents for HCMV in HIV-infected people or vice versa. PMID- 10693652 TI - Importance of the alanine methyl ester side chain for the biological activity profile of dual-function phenyl phosphate derivatives of bromo-methoxy zidovudine. AB - In a systematic search for developing a virucidal spermicide with potent anti human immunodeficiency virus (HIV) and spermicidal activities, we synthesized and evaluated 14 phosphoramidate derivatives of 5-bromo-6-methoxy-zidovudine (PP-BMZ) with differing amino acid ester side chains and para substitutions on the phenyl moiety. Anti-HIV activity was tested by measuring viral p24 antigen production as a marker of viral replication in HIV-1-infected human peripheral blood mononuclear cells. The effect of various PP-BMZ compounds on human sperm motion kinematics was analysed by computer-assisted sperm analysis. Varying the Ala side chain of the phosphoramidate group to other non-polar amino acids, including the cyclic amino acids proline and tryptophan, led to significant alterations in both anti-HIV and spermicidal activities. Our findings highlight the necessity of the Ala side chain and the presence of an electron-withdrawing para-bromo substituent on the phenyl moiety in addition to the bromo-methoxy functional groups on the thymine ring for the PP-BMZ compounds to be effective virucidal spermicides. These membrane permeable dual-function nucleoside analogues may provide the basis for a new strategy aimed at prevention of the sexual transmission of HIV while providing fertility control for women. PMID- 10693653 TI - Prevention of murine influenza A virus pneumonitis by surfactant nano-emulsions. AB - Non-ionic surfactant nano-emulsions have extensive anti-microbial activity and are biocompatible with skin and mucous membranes at effective concentrations. Two nano-emulsion formulations (8N8 and 20N10) made from soybean oil, tributyl phosphate and Triton X-100, were tested for their ability to prevent murine influenza virus pneumonia in vivo. In the initial study, CD-1 mice were administered various dilutions of the nano-emulsions intranasally, and safe dosages and concentrations were determined. Non-toxic concentrations of the nano emulsions were then mixed with influenza virus and applied to the nares of mice. Animals receiving mixtures of two different emulsions (8N8 or 20N10) and a LD50 of virus survived the challenge without evidence of viral infection. To determine if the nano-emulsions could prevent influenza virus infection in vivo when used as a prophylactic treatment, the nano-emulsions (8N8 at 1.0% and 20N10 at 1.0% or 0.2%) were applied to mouse nares 90 min before exposure to 5x10(5) p.f.u./ml virus by nebulized aerosol. Animals pretreated with the nano-emulsions had significantly decreased clinical signs of infection. Only 26.0% (8N8 at 1.0%), 31.25% (20N10 at 1.0%) and 37.0% (20N10 at 0.2%) of animals pretreated with nano emulsion died from pneumonitis, whereas >80.0% of mock pretreated animals succumbed to infection (P<0.005). These findings suggest that non-ionic surfactant nano-emulsions have therapeutic potential for the prevention of influenza virus infection in vivo. PMID- 10693654 TI - A novel peptide aldehyde with activity against human cytomegalovirus in two different in vivo models. AB - Novel peptide aldehydes (PAs) were identified as potent inhibitors of human cytomegalovirus (HCMV) in vitro. Although these compounds were highly effective against HCMV, they did not exhibit any activity against murine cytomegalovirus (MCMV). The purpose of this study was to test the antiviral activity of PA 8 as a representative of this novel class of inhibitors against HCMV in vivo. Because of the strict species specificity of HCMV we had to use two artificial animal models. In the first model, HCMV-infected human cells were entrapped into agarose plugs and transplanted into mice. In the second model, SCID mice were transplanted with human tissues that were subsequently infected with a clinical isolate of HCMV. In these two models the antiviral activity of PA 8 was clearly demonstrated, ganciclovir only being slightly superior in its in vivo antiviral activity. PMID- 10693655 TI - TSAO-T analogues bearing amino acids at position N-3 of thymine: synthesis and anti-human immunodeficiency virus activity. AB - Novel analogues of the anti-HIV-1 lead compound [1-[2',5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl]thymine]- 3'-spiro-5'-(4"-amino-1",2" oxathiole-2',2'-dioxide) (TSAO-T) bearing different amino acids at position N-3 of thymine were prepared and evaluated as inhibitors of HIV replication. The synthesis of the target compounds was accomplished by coupling of the appropriate TSAO intermediate with a conveniently protected (L) amino acid in the presence of BOP and triethylamine, followed by deprotection of the amino acid moiety. Several TSAO derivatives, bearing at N-3 position of the thymine base an L-amino acid retaining the free carboxylic acid, acquired activity against HIV-2, in addition to their inhibitory effect on HIV-1. PMID- 10693656 TI - Antiviral effect of brassinosteroids against herpes virus and arenaviruses. AB - A natural brassinosteroid and a series of synthetic derivatives were found to be good inhibitors of herpes simplex virus type 1 (HSV-1) and arenavirus replication in cell culture. The synthetic compounds tested were analogues of the 24(S) ethylbrassinone. Compounds (22 R,23 R,24S)-2alpha, 3alpha,5alpha,22,23 pentahydroxy-stigmastan-6-one and (22R,23R,24S)-3beta-bromo-5alpha,22,23 trihydroxy stigmastan-6-one were cytotoxic at concentrations of 20-40 microM. (22S,23S,24S)-2alpha,3alpha,22,23-tetrahydroxy-5alpha, stigmastan-6-one, (22R,23R,24S)-3beta-acetoxy-22,23-dihydroxy-5alpha-choles tan-6-one, (22S,23S,24S)-3beta-bromo-22,23-dihydroxy-5alpha-cholestan-6 -one and (22S,23S,24S)-3beta-bromo-5alpha,22,23-trihydroxy-stigmastan -6-one were the most active of the series against HSV-1, with selectivity index (SI) values (CC50/EC50) ranging from 10.6 to 16.5. The majority of the compounds were potent inhibitors of arenaviruses, (22S,23S,24S)-3beta-bromo-5alpha,22,23-trihydroxy stigmastan -6-one being the most active, with SI values of 307.8 and 692.5 for Tacaribe and Junin viruses, respectively. The antiviral activity of brassinosteroid derivatives was not because of direct inactivation; time-of addition experiments suggested that a late step in HSV-1 multiplication was affected, whereas arenaviruses remained susceptible to the compounds throughout the replicative cycle. PMID- 10693657 TI - Predicting inpatient mortality for pediatric trauma patients with blunt injuries: a better alternative. AB - PURPOSE: The aim of this study was to identify significant independent predictors of inpatient mortality rates for pediatric victims of blunt trauma and to develop a formula for predicting the probability of inpatient mortality for these patients. METHODS: Emergency department and inpatient data from 2,923 pediatric victims of blunt injury in the New York State Trauma Registry in 1994 and 1995 were used to explore the relationship between patient risk factors and mortality rate. A stepwise logistic regression model with P<.05 was developed using survival status asthe dependent variable. Independent variables included are elements of the Pediatric Trauma Score (PTS), additional elements from the Revised Trauma Score (RTS), the motor response and eye opening components of the Glasgow Coma Scale (GCS), age-specific systolic blood pressure, the AVPU score, and 2 measures of anatomic injury severity (the Injury Severity Score [ISS] and the International Classification of Disease, Ninth Revision-based Injury Severity Score [ICISS]). RESULTS: The only significant independent predictors of severity that emerged were the ICISS, no motor response (best motor response = 1) from the GCS, and the unresponsive component from the AVPU score. The statistical model exhibited an excellent fit (C statistic = .964). The specificity associated with the prediction of inpatient mortality rate based on the presence of 1 or more of these risk factors was .926, and the sensitivity was .944. CONCLUSION: The best independent predictors of inpatient mortality rate for pediatric trauma patients with blunt injuries include variables not specifically contained in the PTS or the RTS: ICISS, no motor response (best motor response = 1) from the GCS, and the unresponsive component of the AVPU score. PMID- 10693658 TI - Pediatric airbag injuries: the Ohio experience. AB - BACKGROUND/PURPOSE: We sought to determine if properly restrained children, less than 13 years of age, placed in the front passenger seat are at greater risk for trauma from airbag deployment than unrestrained children. METHODS: The charts of children treated at any of 3 regional pediatric trauma centers in Ohio were reviewed for airbag injuries sustained in motor vehicle crashes between January 1995 and September 1998. Injury Severity Scores (ISS) were compared with Mann Whitney Rank Sum Test and outcome data with Fisher's Exact Tests. Statistical significance was set at P< or =.05. RESULTS: Twenty-seven children aged 1 month to 12 years sustained airbag-related injuries. Sixty-one percent were girls. ISS ranged from 1 to 75 with a mean score (+/- SD) of 10 (14.5). All crashes were at reported speeds of less than 45 mph, and 64% were head-on collisions. No significant differences in the mean ISS (P = .074) occurred between groups. Both groups had similar closed head, ocular and facial injuries, extremity fractures, and number of deaths (P = 1.0). Abdominal organ injury was exclusive to the restrained group. Decapitation only occurred among unrestrained children. CONCLUSION: Our data showed that airbags, with or without proper safety restraints, can lead to mortality or serious morbidity in children. PMID- 10693659 TI - Evidence-based guidelines for resource utilization in children with isolated spleen or liver injury. The APSA Trauma Committee. AB - PURPOSE: This study is intended to resolve the disparity and reach consensus on issues regarding the treatment of children with isolated spleen or liver injuries. To maximize patient safety and assure efficient, cost-effective utilization of resources, it was essential to determine current practice. METHODS: Data from the case records of 856 children with isolated spleen or liver injury treated at 32 pediatric surgical centers from July 1995 to June 1997 were collected. The severity of injury was classified by computed tomography (CT) grade and the data analyzed for intensive care unit (ICU) stay, length of hospital stay, transfusion requirement, need for operation, pre- and postdischarge imaging, and restriction of physical activity. Patients with grade V injuries (2.8%) were excluded leaving 832 patients for detailed review. These data and available literature were analyzed for consensus by the 1998 APSA Trauma Committee. RESULTS: Resource utilization increased with injury severity (see Table 2). Based on the data analysis, literature search, and consensus conference, the authors propose guidelines (see Table 3) for the safe and optimal utilization of resources in routine cases. It is important to emphasize that no recommendation falls outside the 25th percentile of current practice at participating centers. CONCLUSIONS: Diversity of treatment, with attendant variation in resource utilization in children with isolated spleen and liver injury of comparable severity is confirmed. This analysis has stimulated a prospective outcomes study with the objective of validating the evidence-based guidelines proposed. This evidence-based study design can bring order and conformity to patient management resulting in optimal utilization of resources while maximizing patient safety. PMID- 10693660 TI - Multiple spontaneous small bowel anastomosis in premature infants with multisegmental necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Fulminant necrotizing enterocolitis (NEC) may result in extensive bowel necrosis. Resection of involved segments may result in short bowel syndrome. Multiple stomas result in complications and further loss of intestinal length with closure. METHODS: Two patients with extensive multisegmental NEC were treated with an intraluminal stent without anastomosis. All necrotic intestine was resected and the remaining viable intestine was lined up over a feeding tube without anastomosis of the intestinal segments. One patient had a diverting jejunostomy and mucous fistula with the tube used to orient the defunctionalized intervening intestinal segments. The second patient had the bowel left in continuity with the tube brought into the jejunem proximal to the first area of resection and distally brought out through the tip of the appendix. Both tubes were brought through the abdominal wall and secured in a loop. RESULTS: Contrast study findings showed that the intestinal segments had auto-anastomosed. In the first case the tube was left in place and intestinal continuity was restored. The patient is now 4 years old and takes full enteral feeds. The latter patient had the enterostomy tube removed at the time of the contrast study, but only tolerated partial feedings and died at 1 year of total parenteral nutrition-related liver failure. CONCLUSION: The technique eliminates nonviable bowel, maximizes length, avoids multiple stomas, and may help avoid reoperation. PMID- 10693661 TI - Diminished epidermal growth factor levels in infants with necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Because epidermal growth factor (EGF) is trophic to the intestinal mucosa, and neonatal necrotizing enterocolitis (NEC) is associated with a disrupted intestinal mucosal barrier, the authors sought to determine whether diminished levels of EGF were present in infants with NEC. METHODS: Saliva, serum, and urine specimens were obtained from infants with NEC during a 3 year period (February 1995 to May 1998). Control patients without NEC were chosen based on similar postnatal age and birthweight. EGF levels were determined by enzyme-linked immunosorbent assay (ELISA). Differences between groups were compared using Mann-Whitney Rank sum test with P less than .05 considered significant. Results are presented as mean values +/-SEM. RESULTS: Twenty-five infants with NEC were compared with 19 control patients. Birth weight (1,616+/ 238 g control v. 1,271+/-124 g NEC) and postnatal age (23+/-6 days control v. 22+/-3 days NEC) were similar. Infants with NEC had significantly lower levels of EGF in both saliva (590+/-80 pg/mL control v. 239+/-41 pg/mL NEC; P<.001) and serum (35+/-8 pg/mL control v. 5.6+/-1.9 pg/mL NEC; P<.001). Urinary EGF was also lower in the NEC group, but was not statistically significant. CONCLUSIONS: Premature infants with NEC have significantly diminished levels of salivary and serum EGF. Reduced levels of this growth factor may distinguish infants at risk for NEC and play a pivotal role in the pathogenesis of the perturbed intestinal mucosal barrier that is central to this condition. PMID- 10693662 TI - Erythropoietin and the incidence of necrotizing enterocolitis in infants with very low birth weight. AB - BACKGROUND/PURPOSE: The presence of erythropoietin (Epo) in human milk and the expression of Epo receptors on intestinal villous enterocytes of neonates suggest that Epo has a role in growth and development of the gastrointestinal tract. On this basis, the authors hypothesized that recombinant Epo (rEpo) given for prevention or treatment of the anemia of prematurity would protect against necrotizing enterocolitis (NEC). METHODS: A retrospective cohort study was conducted from a university neonatal intensive care unit of 483 very low birth weight (500 to 1,250 g) neonates born from July 1, 1993 to January 1, 1998. RESULTS: A total of 260 neonates received rEpo, and 223 did not (control group). The rEpo and control groups were similar in gender distribution (52% v. 48% boys), gestational age (26.8+/-2.1 v. 27.6+/-2.9 weeks; mean +/- SD), birth weight (895+/-198 v. 911+/-208 g), 1 and 5 minute Apgar scores (4.2 and 6.1 v4.7 and 6.7), and incidence of severe intraventricular hemorrhage (8.9% v. 10.3%). The rEpo group had a lower incidence of NEC (12 of 260, 4.6% v. 24 of 223, 10.8%; P = .028, 95% confidence interval for difference: -0.108 to -0.015). CONCLUSION: In very low birth weight infants, the incidence of NEC is lower in those who received rEpo. PMID- 10693663 TI - Myofibroblast induction with transforming growth factor-beta1 and -beta3 in cutaneous fetal excisional wounds. AB - BACKGROUND/PURPOSE: In a noncontractile fetal rabbit model, the authors recently have shown the induction of excisional wound contraction with sustained-release cellulose implants formulated with transforming growth factor (TGF)-beta. The purpose of this study was to test the hypothesis that the excisional wound contraction in this model is associated with the induction of myofibroblasts in the surrounding dermis, demonstrated by the presence of alpha-smooth muscle actin. METHODS: Cellulose discs were formulated with either 1.0 microg of TGF beta1 (n = 6); 1.0 microg of TGF-beta3 (n = 9); 10 microg of TGF-beta3 (n = 6); or their carrier protein, bovine serum albumin (BSA; n = 9), for sustained release over 5 days. Each disc was implanted into a subcutaneous pocket on the back of a fetal New Zealand White rabbit in utero on day 24 of gestation (term, 31 days). A full-thickness, 3-mm excisional wound (7.4 mm2) was then made next to the implanted cellulose disc. All fetuses were harvested at 3 days. The amount of alpha-smooth muscle (SM) actin in the dermis around the implants and wounds was determined using immunohistochemical techniques. RESULTS: Excisional wounds exposed to 1.0 microg of TGF-beta1 (5.6+/-2.0 mm2), 1.0 microg of TGF-beta3 (6.9+/-1.0 mm2), and 10 microg of TGF-beta3 (2.7+/-1.0 mm2) were significantly smaller when compared with the BSA control group (12.8+/-1.1 mm2; P<.05). Furthermore, there was a significant increase in staining for alpha-SM actin in the TGF-beta1 (1.8+/-0.5) and 10 microg TGF-beta3 (2.8+/-0.2) groups in comparison with the scant staining in the BSA control group (0.5+/-0.2; P<.05). CONCLUSIONS: TGF-beta1 and -beta3 induce alpha-SM actin and contraction of cutaneous excisional wounds in a fetal noncontractile model. This model of inducible cutaneous excisional wound contraction may be useful in further determining the role of the myofibroblast in wound contraction and the physiology underlying this poorly understood aspect of wound healing. PMID- 10693664 TI - TNP-470 inhibits intraabdominal adhesion formation. AB - BACKGROUND/PURPOSE: Angiogenesis plays an integral role in wound healing and tissue remodeling. The authors hypothesized that inhibition of angiogenesis would reduce intraabdominal adhesion formation. METHODS: In 98 C57BL6/J mice, a 2-cm midline laparotomy was performed and a 5 mm2 SILASTIC (Dow Corning, Midland, MI) patch fixed to the right side of the peritoneum. Mice were injected with normal saline (n = 54) or TNP-470, an inhibitor of angiogenesis (n = 44; 30 mg/kg every other day over 6 days before surgery until 10 days after surgery). Animals were killed on postoperative days 10, 15, 35, and 55. Adhesions to the SILASTIC (Dow Corning) patch were scored based on their extent, type, and tenacity. Angiogenesis was quantified digitally as the area of vascularized peritoneum over the patch. RESULTS: At day 10, when TNP-470 was stopped, the percentage of vascularized peritoneum over the patch was less in treatment animals than in controls (P = .004). At day 35, the patch in treatment animals was completely covered by vascularized peritoneum, similar to controls. Adhesions in TNP-470 animals were reduced at day 10 compared with controls (P<.05) and remained reduced off treatment at day 55. CONCLUSIONS: Angiogenesis appears to play an important role in the development of intraabdominal adhesions, because the extent of early neovascularization correlates with adhesion formation. Perioperative treatment with TNP-470, a potent endothelial cell inhibitor, reduced vessel ingrowth over the patch and was associated with a sustained reduction in adhesion formation. PMID- 10693665 TI - Extracorporeal life support outcome for 128 pediatric patients with respiratory failure. AB - PURPOSE: The aim of this study was to describe a single-center experience with pediatric extracorporeal life support (ECLS) and to determine variables predictive of outcome in pediatric patients, both before the institution of ECLS and while on support. METHODS: From October 1985 to September 1998 the authors supported 128 children with severe acute hypoxemic respiratory failure(n = 121, Pao2/FIo2 ratio = 58+/-29) or acute hypercarbic respiratory failure (n = 7, Paco2 = 128+/-37), despite maximal conventional ventilation. Mode of access included venoarterial bypass (VA, n = 64), venovenous bypass (VV, n = 53), and VV to VA bypass (n = 11). The techniques used included lung rest, pulmonary physiotherapy, diuresis to dry weight using hemofiltration if needed, minimal anticoagulation, and optimal systemic oxygen delivery. RESULTS: The median age was 1.4 years (range, 2 weeks to 17 years). The mean duration of ECLS was 288+/-240 hours (range, 4 to 1148 hours or 0.2 to 47.8 days). Lung compliance increased from 0.32+/-0.02 mL/cm H2O/kg to 0.59+/-0.03 mL/cm H2O/kg in survivors, but only increased from 0.34+/-0.02 mL/cm H2O/kg to 0.35+/-0.02 mL/cm H2O/kg in nonsurvivors (P<.002 comparing change between survivors and nonsurvivors). Mean body weight decreased from 9%+/-2% over dry weight to 4%+/-2% in survivors, whereas in nonsurvivors the mean body weight increased from 25%+/-5% over dry weight to 35%+/-7% (P<.001). Outcome results by diagnosis were pneumonia, 73%; acute respiratory distress syndrome, 67%; and airway support, 60%, with overall lung recovery occurring in 77%, and hospital survival in 71%. Multivariate logistic regression modelling of patients with hypoxemic respiratory failure found the only pre-ECLS variable significantly associated with outcome to be pH (P<.05). Variables during the course of ECLS significantly associated with decreased survival were the presence of creatinine greater than 3.0 (P<.01), the need for inotropes (P<.04), failure to return the patient to dry weight (P<.04), and lung compliance that did not improve significantly. (P<.01). CONCLUSIONS: ECLS provides life support in severe respiratory failure in children, allowing time for injured lungs to recover. Pre-ECLS predictors, such as pH and variables during ECLS, such as presence of renal failure, improvement in compliance, return to dry weight, and the need for inotropes on ECLS, may be useful for predicting outcome. PMID- 10693666 TI - Antenatal dexamethasone enhances endothelin receptorB expression in hypoplastic lung in nitrofen-induced diaphragmatic hernia in rats. AB - BACKGROUND/PURPOSE: The hypoplastic lung and persistent pulmonary hypertension (PPH) are the principle causes of high mortality and morbidity in infants with congenital diaphragmatic hernia (CDH). Endothelin-1 (ET-1), which is produced by vascular endothelial cells and some leukocytes, plays a key role in modulating pulmonary vascular tone in PPH. Two different receptors (ET(A) and ET(B)) for ET 1 have been characterized. Binding of ET-1 to ET(A), which is present on smooth muscle cells in fetal lung, results in vasoconstriction. However, binding of ET-1 to ET(B), which is present on endothelial cells results in vasodilation mediated by endogenous nitric oxide. Antenatal glucocorticoid therapy has been shown to prevent abnormal pulmonary arterial structural changes in animal model with CDH. The aim of this study was to investigate the effect of antenatal glucocorticoid administration on ET-1 system in nitrofen-induced CDH hypoplastic lung in rats. METHODS: A CDH model was induced in pregnant rats after administration of nitrofen on day 9.5 of gestation. Dexamethasone (Dex) was given intraperitoneally on days 18.5 and 19.5 of gestation. Cesarean section was performed on day 21 of gestation. Rat ET-1 protein expression was measured in solubilized lung tissue extracts, by sandwich type enzyme-linked immunosorbent assay (ELISA) analysis. Reverse transcription polymerase chain reaction was performed to evaluate the relative amount of ET-1, ET(A), and ET(B) mRNA expression. RESULTS: The ET-1 protein and mRNA expression of ET-1 and both receptors were increased significantly in CDH lung compared with controls. Although there was no significant difference in ET(A) mRNA expression between CDH lung with Dex treatment and without Dex treatment, ET(B) mRNA expression was elevated significantly in CDH lung with Dex treatment compared with CDH lung without Dex treatment. CONCLUSION: These findings suggest that antenatal glucocorticoid therapy may modulate pulmonary vascular tone in CDH hypoplastic lung by selectively upregulating local expression of ET(B). PMID- 10693667 TI - Remodeling of pulmonary arteries in human congenital diaphragmatic hernia with or without extracorporeal membrane oxygenation. AB - PURPOSE: The aim of this study was to describe in detail the perinatal developmental profile of the pulmonary vasculature in congenital diaphragmatic hernia (CDH) and to examine the potential beneficial effects of extracorporeal membrane oxygenation (ECMO) on the vascular morphology. Additionally the authors aimed to identify the differences in pulmonary vascular morphology among CDH cases according to the primary cause of death: either extreme lung hypoplasia (LH) or persistent pulmonary hypertension (PPH). METHODS: The authors studied autopsy sections from 30 high-risk CDH cases with respect to the pulmonary arteries in relation to gestational age (GA) and ECMO treatment. They were grouped into CDH-I: 20 cases with GA greater than 34 weeks who were not subjected to ECMO and CDH-II: 10 cases with GA greater than 34 weeks, who were subjected to ECMO for an average time of 237 hours. Five age-matched neonates who died from placental insufficiency or birth asphyxia without evidence of lung hypoplasia served as controls (CON). Medial and adventitial thicknesses of pulmonary arteries were measured in lung sections stained with Elastic van Gieson by 2 investigators blinded for the clinical data. Immunohistological staining with anti-alpha-smooth muscle actin (alpha-SMA) was performed to confirm the precise location of the arterial media before morphometry. CDH cases were subgrouped and compared according to the primary cause of death. Unpaired Student t test was used for statistics, with significant P value < or =.05. RESULTS: In CDH newborns, a significant increase in medial, adventitial, and total wall thickness was found in pulmonary arteries with an external diameter of less than 200 microm as compared with age-matched controls (P<.004, .0001, and .0009, respectively). ECMO-treated CDH newborns showed a significantly thinner arterial adventitia than CDH patients who did not receive this treatment (P<.0001), approaching normal values. However, the medial thickness remained increased. Morphometrically, no significant differences in CDH cases between patients dying of PPH or severe LH could be determined. CONCLUSIONS: (1) In CDH, there is failure of the normal arterial remodeling processes occurring in the perinatal period. (2) Pulmonary vascular morphology in CDH does not differ between the groups with lung hypoplasia or persistent pulmonary hypertension as primary cause of death. (3) Adventitial thinning of these arteries might be one of the mechanisms by which ECMO alters PPH in CDH cases. PMID- 10693668 TI - Lung growth induced by tracheal occlusion in the sheep is augmented by airway pressurization. AB - BACKGROUND/PURPOSE: Prenatal tracheal occlusion (TO) has been shown to accelerate lung growth, yet the mechanism for this effect is poorly understood. Increased intratracheal pressure (ITP) with accumulation of lung fluid and secondary airway distension (stretch) may provide a mechanical stimulus for growth. In this study, ITP after TO is measured continuously, and the effect of altering ITP on lung growth is examined. METHODS: Fetal lambs of 115 to 120 days of gestation (term, 145 days) underwent placement of an intratracheal catheter and an amniotic fluid reference catheter. First, ITP was monitored continuously in normal controls (n = 4) and in fetuses undergoing TO (n = 6). In a subsequent study, 2 groups of fetuses were compared. In the TO group (n = 5) ITP was monitored after TO. In the pressurized group (n = 5) ITP was maintained at 7 to 8 mm Hg by a continuous servo regulated pump that maintains a preset pressure by lactated Ringers infusion. The animals were killed after 4 days, and lung growth was compared. RESULTS: In the control animals, ITP remained constant at 0.4 to 1.5 mm Hg. In the TO animals, ITP increased gradually during the initial 24 hours and plateaued at 4 to 5 mm Hg. In the second set of animals, ITP in the pressurized group was maintained at approximately 8 mm Hg using the infusion system. Lung volume (135.7+/-17.4 v. 95.2+/-14.8 mL/kg; P<.01), lung weight to body weight (6.70+/ 0.73 v. 5.33+/-0.77%; P<.05), whole right lung dry weight (3.10+/-0.22 v. 2.63+/ 0.20 mg/kg; P<.05), and right lung DNA and protein contents (87.3+/-6.0 v. 74.6+/ 8.1 mg/kg, 2,310+/-248 v. 1,860+/-196 mg/kg, respectively; P<.05) were increased significantly in the pressurized group compared with the TO group. Morphometry confirmed greater volume of respiratory region and increased alveolar surface area in the pressurized lung. CONCLUSIONS: TO results in a gradual increase in ITP over 15 to 24 hours, which plateaus at 4 to 5 mm Hg. Further increasing ITP by infusion of crystalloid significantly augments lung growth beyond that observed with TO alone. These data support the hypothesis that airway pressure and secondary mechanical stretch are the primary stimuli of TO induced lung growth. PMID- 10693669 TI - Adrenocortical insufficiency in infants with congenital diaphragmatic hernia: a pilot study. AB - BACKGROUND/PURPOSE: Prior reports have documented that premature infants do not have normal serum levels of cortisol. In contrast, full-term infants usually have adequate cortisol levels. The stress response in critically ill infants may be vital to their recovery. The purpose of this pilot study was to determine whether critically ill full-term infants with congenital diaphragmatic hernia (CDH) show a subnormal adrenal stress response. METHODS: Random serum cortisol levels in infants with CDH (n = 10) were measured using fluorescent polarization immunoassay. In addition, serum cortisol levels were measured after exogenous adrenocorticotropic hormone stimulation (Cosyntropin stimulation test). RESULTS: Six of the 10 infants studied died. Most (79%) of the cortisol levels were subnormal (<7 microgm/dL). Although no significant differences in mean cortisol levels from terminally ill infants compared with surviving infants were detected, survivors tended to have higher cortisol levels. Cosyntropin stimulation resulted in inappropriately low cortisol levels in 2 of the 4 fatally ill patients tested (<30 microgm/dL) and normal responses in the 2 survivors tested. CONCLUSIONS: Infants born with CDH may have an inadequate adrenal stress response despite a life-threatening anomaly. A large-scale prospective study may be warranted to confirm this apparent association. Corticosteroid therapy may be beneficial in this population of patients. PMID- 10693670 TI - New mouse models of congenital anorectal malformations. AB - BACKGROUND/PURPOSE: The genetic, embryological, and pathogenetic aspects of hindgut development remain poorly understood. Recently, the morphogenetic pathway involving the Sonic hedgehog (Shh) gene has been shown essential to the normal development of many midaxial organs, including the foregut. This study reports genetically based murine models of congenital anorectal malformations (CAM) involving the Shh-responsive transcription factors, Gli2 and Gli3. Its purpose is to show the necessity of these 2 factors to normal hindgut development. METHODS: Gli2-/- mutants were generated by a targeted deletion. Gli3-/- mutants are spontaneous mutants involving the Gli3 gene. Gli2-/- Gli3+/- mutants were generated by intercrossing double heterozygotes. Whole-mount midsagittal sections of the embryos were analyzed on embryonic days (E) 11.5 and E13.5. RESULTS: Gli3 /- mutants had anal stenosis and ectopic anus, and Gli2-/- mutants showed imperforate anus and rectourethral fistula. Gli2-/- Gli3+/- mutants had a cloacal abnormality. CONCLUSIONS: The phenotypic abnormalities observed in these mutant mice are identical to the spectrum of human CAM. The severity of the phenotype appears to reflect the gene dose. Gli2 and Gli3 play an important role in the normal development of murine hindgut. The results of this study provide, for the first time, a molecular basis for CAM. PMID- 10693671 TI - Home bowel preparation for elective colonic procedures in children: cost savings with quality assurance and improvement. AB - BACKGROUND/PURPOSE: The current health care environment pressures providers to lower cost and demands quality care that is measured by outcomes and patient satisfaction. Most insurers will not approve bed days for in-hospital preoperative bowel preparations for elective colorectal procedures. This policy does not take into account that infants and children are unable to tolerate large volumes of enteral preparation, which adversely affects outcome because of an inadequate preparation. This report describes a prospective evaluation of a standard home bowel preparation regimen utilizing local and regional home health care agency support. METHODS: For an elective colorectal procedure, pediatric patients underwent a home bowel preparation using GoLYTELY (100 mL/kg) via a nasogastric tube infused over 4 hours by a pediatric home health nurse trained in this technique. During the bowel preparation, the nurse educated the family members about the service and performed physiological monitoring to insure safety. At the completion of the preparation, any unusual events were transmitted to the staff surgeon for further instructions. Our initial 30 patients were treated by our hospital home health agency personnel to insure safety. Since then, 41 additional bowel preparations have been performed by statewide agencies. RESULTS: Seventy-one patients underwent complete home bowel preparation (45 boys; 26 girls). The age range was 3 months to 9 years (average, 5 months). There was one complication caused by incorrect mixing of GoLYTELY causing gastrointestinal cramping. All 71 home bowel preparations were recorded as good at the time of the colorectal procedure by the staff pediatric surgeon. The average cost for home bowel preparation was $300 in network, and $350 out of network. This compares with an inpatient hospital day cost of greater than $800 ($36,000 savings). CONCLUSIONS: This technique offers the pediatric surgeon an opportunity to maintain a high standard of quality care while using home health agency personnel to minimize cost. This program is safe, effective, and associated with a good outcome and a high degree of family satisfaction. PMID- 10693672 TI - Transanal mucosectomy in the treatment of Hirschsprung's disease. AB - BACKGROUND/PURPOSE: Transanal mucosectomy of the aganglionic segment of colon is a critical step in minimally invasive surgery for Hirschsprung's disease. The purpose of this study was to examine the outcome of patients undergoing transanal mucosectomy. METHODS: From January 1979 to November 1998, 26 patients (ages 25 days to 17 years) underwent transanal mucosectomy for Hirschsprung's disease. Seventeen (65%) had partial transanal mucosectomy (PTM; 1979 to 1998) and 9 (35%) complete transanal mucosectomy (CTM; 1995 to 1998). In PTM, a 2- to 3-cm mucosal dissection was begun 1 cm above the dentate line in conjunction with transabdominal endorectal dissection (modified Soave). In CTM, the entire mucosal dissection was performed transanally as part of a laparoscopically assisted Soave procedure. Results were obtained by chart review and personal communication. Patients were assessed clinically for continence where age appropriate (>3 years) and for development of constipation, postoperative enterocolitis, and anal stricture. RESULTS: One of 16 (6.2%) of the PTM group was incontinent versus none (4 patients) in the CTM group. Five of 17 (29.4%) of the PTM group were constipated versus 4 of 9 (44.4%) in the CTM group (t test, P = not significant). Postoperative enterocolitis developed in 4 of 17 (23.5%) of the PTM group versus 6 of 9 (66.6%) in the CTM group (t test, P<.05). Three of 6 (50%) of the CTM group versus none in the PTM group required hospitalization for bowel rest, rectal washouts, and antibiotics. All patients were well at the time of the report. Anal stricture was not seen in either group. CONCLUSIONS: Constipation and postoperative enterocolitis are a significant feature of transanal mucosectomy for Hirschsprung's disease deserving close surveillance, especially in patients in whom the entire mucosal dissection was performed transanally. Continence appears to be satisfactorily preserved from these preliminary results. PMID- 10693673 TI - Historical changes in the postoperative treatment of appendicitis in children: impact on medical outcome. AB - BACKGROUND/PURPOSE: The introduction of managed care in the 1980s caused increased pressure to reduce costs for hospitalized patients. The authors hypothesized that these market forces have resulted in a decreased hospital stay and utilization of sophisticated diagnostic testing in children treated for appendicitis. If true, the impact of this paradigm shift on patient outcome is unknown. METHODS: Hospital records for 913 pediatric patients treated for appendicitis from 1974 to 1998 were reviewed retrospectively. Patients were stratified into those with perforated appendicitis (PA) and nonperforated appendicitis (NPA). Demographics, perioperative hospital course, diagnostic testing, complications, and long-term outcomes were analyzed after stratification into time intervals. RESULTS: Over time, children with NPA were treated with shorter antibiotic courses (P<.05) and were placed on a regular diet earlier (P<.05). These changes in treatment resulted in an earlier discharge (P<.05). The amount of time to become afebrile with a normal white blood cell count (WBC) did not change over time. Children with PA exhibited similar results with shorter antibiotic courses (P<.05), earlier dietary intake (P<.05) and earlier hospital discharge (P<.05) over time. In all children with appendicitis there was no significant difference in the rate of wound infections, abscesses requiring drains, readmission, or reoperations overtime. The utilization of abdominal radiographs (83%) and ultrasonography (USN; 40%) was high and remained unchanged over time. Utilization of computed tomography (CT scan) was low (4.3%) in the early decades and was not used as a preoperative test from 1991 to 1994. Given the high diagnostic accuracy of a pediatric surgeon for this disease, Bayesian analysis indicates that USN utilization rates should be 15%. CONCLUSIONS: The market pressures of managed care have resulted in a new treatment paradigm with an earlier discharge of all children with appendicitis. There has been no concomitant increase in the complication rate in either group as a result of this paradigm shift. Bayesian analysis indicates that USN and abdominal radiographs are overutilized in our institution. PMID- 10693674 TI - Is the grass greener? Early results of the Nuss procedure. AB - BACKGROUND/PURPOSE: Minimal access surgery (MIS, Nuss Procedure) is gaining acceptance rapidly as the preferred method for pectus excavatum repair. This shift in operative management has followed a single institution's evaluation of the procedure. This report describes an additional experience with the Nuss procedure. METHODS: Twenty-one patients with pectus excavatum underwent repair by the Nuss Procedure. The patients ranged in age from 5 to 15 years (average, 8.2 years). There were 19 boys and 2 girls. RESULTS: In 1 patient (age 5 years) the MIS procedure was aborted because of persistence of chest wall asymmetry. The other 20 patients had completion of their procedure without intraoperative complication. The operating times ranged from 45 to 90 minutes; however, there was an additional anesthetic set-up time (average, 45 minutes). All cases utilized a single support bar (11 to 17 inches). Patients underwent extubation in the operating room and were admitted to a ward bed with an epidural catheter in place for pain control and received intravenous analgesia. The hospital stay ranged from 4 to 11 days and averaged 4.9 days. Early postoperative complications included ileus (n = 1), bilateral pleural effusion (n = 2), atelectasis (n = 1), fungal dermatitis (n = 1), pneumothorax (n = 1), and flipped pectus bar (n = 2). Delayed complications included flipped pectus bar (n = 2), marked pectus carinatum requiring bar removal (n = 1), mild carinatum (n = 1), mild bar deviation (n = 1), progressive chest wall asymmetry (n = 3) with 1 requiring bar removal and open pectus repair, pleural effusion (n = 1), and chronic persistent pain requiring bar removal (n = 1). The length of follow-up is 3 to 20 months with an average of 12.3 months. CONCLUSIONS: The Nuss Procedure is quick, minimally invasive, and a technically easy method to learn; however, our data indicate there is a significant learning curve. Although previous reports suggest that few complications occur, we believe further assessment of patient selection regarding age, presence of connective tissue disorder, and severe chest wall asymmetry are still needed. Long-term follow-up also will be required to assure both health professionals and the public that this is the procedure of choice for patients with pectus excavatum. PMID- 10693675 TI - Outcome analysis of minimally invasive repair of pectus excavatum: review of 251 cases. AB - BACKGROUND/PURPOSE: Since the first report in 1997 by Dr Nuss of the technique for minimally invasive repair of pectus excavatum (MIRPE), the popularity and demand for this operation has increased dramatically. Many pediatric surgeons became familiarized with MIRPE and have applied it to a large number of patients. Outcomes and complications have not yet been defined. METHODS: A comprehensive survey of APSA members was conducted to review technical problems, complications, and outcomes of this new technique. RESULTS: Of the 74 survey responders, 31 (42%) currently use the MIRPE as their procedure of choice, and 251 cases were reviewed. A total of 74.2% of surgeons relied on direct observation and written documentation to obtain training in MIRPE. Less than 60% used the chest index in the preoperative assessment. A total of 98% used the Walter Lorenz bar for the MIRPE. The most common complication was bar displacement or rotation requiring reoperation (9.2%). Pneumothorax requiring tube thoracostomy was reported in 4.8%. Less common problems included infectious complications (2%), pleural effusion (2%), thoracic outlet obstruction (0.8%), cardiac injury (0.4%), sternal erosion (0.4%), pericarditis (0.4%), and anterior thoracic artery pseudoaneurysm (0.4%). Three patients (1.2%) required early strut removal. Reoperation using the open modified Ravitch approach was performed in 2 patients (0.8%). Most surgeons indicated that teenaged patients (>15 years old) were at higher risk for complications. Thoracoscopy in combination with MIRPE was used by 61% of the surgeons. Overall patient satisfaction was rated as excellent or good (96.5%). CONCLUSIONS: The relatively high incidence of problems with MIRPE is probably related to the learning curve associated with the introduction of this new technique. Awareness of technical details, careful patient selection, use of a stabilizing bar, and thoracoscopy likely will result in decreased complications. Long-term results are yet to be determined. The development of a national registry is of great importance for further outcome analysis of MIRPE. PMID- 10693676 TI - Slide tracheoplasty for congenital tracheal stenosis: a case report. AB - BACKGROUND/PURPOSE: A variety of techniques have been used to manage pediatric congenital tracheal stenosis. The authors report the technique of slide tracheoplasty for a child with long congenital tracheal stenosis. METHODS: A 2 year-old male presented with a history of stridor with feeding. Bronchoscopy findings showed 50% stenosis from complete cartilaginous rings, extending from 2.5 cm below the vocal cords to 2 cm above the carina. Through a neck incision, the trachea was exposed from the cricoid to both bronchi and transected at the midpoint of the stenosis. The upper trachea was split anteriorly to the area of stenosis just below the cricoid. The lower trachea was split posteriorly in the midline. Posterior dissection allowed sliding and anastomosis of both tracheal segments while the lateral vascular supply was left intact. A brace was placed to maintain cervical flexion, and the patient underwent extubation in the operating room. RESULTS: He recovered without complication and was dis charged on postoperative day 4. CONCLUSION: Slide tracheoplasty offers several advantages for tracheal reconstruction because it is performed with the native tracheal tissues, can be accomplished through a transverse collar incision, and can repair long stenoses without significant tracheal shortening. PMID- 10693677 TI - A new method of treatment for complete tracheal rings in an infant: endoscopic laser division and balloon dilation. AB - PURPOSE: The authors describe a new technique for management of complete tracheal rings in infants. METHODS: The procedure consists of rigid bronchoscopy with KTP laser division, in the posterior midline, of the complete rings and gradual advancement of the bronchoscope aided by endoscopic balloon dilation. CONCLUSIONS: The laser division, coupled with balloon dilation, allows for controlled separation of the cartilages posteriorly. The anterior esophageal wall buttresses the posterior tracheal separation. PMID- 10693678 TI - Management of parapneumonic collections in infants and children. AB - BACKGROUND/PURPOSE: Video-assisted thoracoscopic surgery (VATS) has a recognized role in treatment of empyema thoracis. The purpose of this report is to show the value of initial VATS as the primary treatment of parapneumonic collections. METHODS: A retrospective review was done of 139 children who required surgical consultation for parapneumonic collections between January 1992 and July 1998. Management options were (M1) thoracentesis, chest tube drainage, or fibrinolytic therapy and delayed thoracotomy for unresolved collections; (M2) thoracentesis, chest tube drainage, fibrinolytic therapy with delayed VATS if the child remained ill; or (M3) primary VATS. Comparative data included age, duration of prehospital illness, oxygen requirements, white blood cell count, bacterial culture results, number of procedures performed per patient, duration of chest tube drainage, complications, and length of stay. Kruskal-Wallis 1-way analysis was used, with significance at P less than .05. RESULTS: A total of 60 children were treated by M1, 38 by M2, and 41 by M3. Age, duration of prehospital illness, oxygen requirements, white blood cell count, bacterial culture results, and complication rates were comparable. The median length of stay was 12 days for M1, 11 days for M2, and 7 days for M3, with M3 significantly shorter at P<.001. The number of procedures was a median of 2 in M1, 2 in M2, and 1 in M3, with M3 significantly fewer at P<.001. Duration of chest tube drainage was a median 5 days for M1 and 3 days for M2 and M3, with M1 significantly longer at P<.001. There were 9 thoracotomies in the M1 group, 3 in the M2 group, and none in the M3 group. One child in M3 required a second VATS. CONCLUSIONS: Primary VATS has significantly decreased the number of procedures, duration of chest tube drainage and length of stay for children with parapneumonic effusions. Primary VATS appears to be of value in management of bacterial pneumonia with effusion. PMID- 10693679 TI - Thoracoscopic lung resection in children. AB - PURPOSE: The aim of this study was to evaluate the technique of video-assisted thoracic surgery (VATS) in lung resections in infants and children. METHODS: From December 1992 to December 1998 113 consecutive patients, ages 3 weeks to 19 years, underwent VATS for biopsy or resection of various lung pathology. This included 88 wedge biopsies, 12 resections of bullous or cystic disease, 9 lobectomies or segmental resections, and 4 bronchogenic cysts. RESULTS: All procedures were completed successfully. Two patients with metastatic disease had surgery converted to a standard thoracotomy for extensive resections. The average operating time for a wedge biopsy of 2 sites was 26 minutes and 210 minutes for a lobectomy. The average hospital stay after wedge resection was 1.1 days. There were no complications related to the VATS approach. CONCLUSION: VATS is a safe and effective technique in the diagnosis and treatment of pediatric pulmonary disease. PMID- 10693680 TI - The 'penny pincher': a new technique for fast and safe removal of esophageal coins. AB - BACKGROUND/PURPOSE: Considerable debate surrounds the choice of technique for the removal of esophageal coins: endoscopic extraction versus dislodgement with a Foley balloon versus dislodgement using bougienage. The "penny-pincher" (PP) technique was developed as an alternative, incorporating the main advantages of these various approaches. METHOD: The PP technique is based on the insertion of a fluoroscopically guided device that consists of a grasping endoscopic forceps covered by a soft rubber catheter. The forceps provides a firm hold on the coin. The catheter protects the oropharynx and aligns the device with the coin. Once the tip of the catheter is close to the upper edge of the coin, the previously retracted radiopaque prongs of the grasping forceps are deployed and the edge of the coin firmly grasped and extracted. The procedure is done without anesthesia or sedation. RESULTS: Twenty coins were removed from 19 consecutive children with a mean age of 34 months. Average lip-to-lip removal (including fluoroscopy) time was 41 seconds. There were no complications, and all patients were discharged shortly after coin removal. CONCLUSION: The penny-pincher method for the removal of upper esophageal coins combines the simplicity, speed, and cost effectiveness of balloon catheter or bougie coin dislodgement with the safety and secure grasping of endoscopic or forceps removal. PMID- 10693681 TI - Preoperative home care for esophageal atresia--a survey. AB - BACKGROUND/PURPOSE: Long gap esophageal atresia may require months of preoperative management before definitive repair. When 2 recent patients prompted the authors to consider preoperative home care, no published protocol could be identified. This survey is undertaken to determine pediatric surgeons' experience with preoperative home care for long gap atresia. METHODS: A total of 543 surgeons were asked if any patients with long gap atresia had been treated preoperatively at home. For patients sent home, information on nursing care, insurance issues, complications, and timing or type of repair was requested. RESULTS: A total of 380 surveys (70%) were returned. A total of 165 surveys representing 348 patients were included. Forty-one of 165 surgeons (25%) treated 63 of 87 patients (72%) with long gap atresia and an intact upper pouch at home. Home nursing care was provided for 44 patients (70%): 16 (36.4%) night shift, 2 (4.5%) day shift, 3 (6.8%) 24 hour, and 23 (52.3%) intermittent care. No complications referable to preoperative home care were reported. CONCLUSIONS: Significant hesitancy and practice variance exists regarding preoperative home care of patients with long gap esophageal atresia. Many surgeons are satisfied with the safety and cost effectiveness of this technique, although a prospective, multicenter trial is needed to study it in a randomized, controlled fashion. PMID- 10693682 TI - Split-liver transplantation: a comparison of ex vivo and in situ techniques. AB - BACKGROUND/PURPOSE: The expanding applicability of liver transplantation as treatment for end-stage liver disease has fostered a disproportionate increase in liver transplant candidates in the face of an unchanging pool of donor organs. This has resulted in disparities in pretransplant waiting times and deaths. The splitting of a liver allograft allows for the transplantation of 2 recipients, usually an adult and a child, thus providing a means to expand the cadaveric donor pool. METHODS: The authors present their results on the performance of an ex vivo (back table) split and in situ (in a hemodynamically stable cadaveric donor) split to evaluate safety, applicability, and effectiveness. Between November 1989 through April 1998, 54 split-liver transplant recipient operations were performed (24 pediatric and 30 adult). Thirty donors were procured: the ex vivo splitting yielded 25 grafts from 13 donors (donor age, 24.6+/-11 years), and the in-situ technique yielded 29 grafts from 17 donors (mean donor age of 25.5+/ 10.4 years). Five donors involved interinstitutional sharing for which the left side of the graft was kept at the host hospital and the right side grafts were utilized at our center. RESULTS: Overall 1-year patient survival was 85%, with a graft survival of 72%. Patient survival was similar with ex vivo (74%) as compared with the in situ splitting group (96%; P = .06), as was graft survival in ex vivo (61 %) versus in situ (81%) splitting (P = .15). The pediatric population benefited most from the in situ technique, with a 1-year patient survival rate of 100% with the in situ technique versus the ex vivo technique survival rate of 64% at 1 year (P = .02). The 1-year graft survival comparing these 2 techniques was 83% for the in situ group versus 45% for the ex vivo group. Analysis of the program evolution of split-liver transplantation suggested a time-dependent learning curve, which was applicable to surgical splitting technique, implantation, and recipient selection. CONCLUSIONS: The principle of splitting livers from cadaveric donors is fundamentally sound and technically feasible. The authors' outcomes analysis using 2 different procurement techniques suggests that the in situ technique is clinically efficacious, can be used alternatively with the ex vivo technique, and is comparable to whole-liver allograft transplantation. PMID- 10693683 TI - Composite liver--small bowel allografts with preservation of donor duodenum and hepatic biliary system in children. AB - BACKGROUND/PURPOSE: Liver and intestinal transplantation is commonly required for children with intestinal failure who suffer concomitant total parenteral nutrition (TPN)-induced liver failure. Retrieval of such composite allografts using previously described "standard techniques" mandates reconstruction of the biliary system with a defunctionalization loop of the proximal allograft jejunum. The occasional posttransplant biliary complications have been associated with significant morbidity and mortality. Also, size matching has limited the pool of donor organs for this patient population. To improve outcome and increase the donor pool the authors have utilized a duodenal-sparing composite liver small bowel allograft technique (DLSBTx) by preserving the head of the pancreas and the pancreatic-duodenal arteries. This precludes a biliary drainage procedure. METHODS: Nine children (5 girls, 4 boys), with a mean age of 1.4 years (range, 1 to 17.4 years) received a DLSBTx. In 2 patients the liver was reduced; 1 patient received the whole pancreas. The mean recipient weight at the time of transplantation was 17.4 kg (range, 6.6 to 49.8 kg). The mean age and mean weight for donors was 7.9 years (range, 3 days to 22 years) and 25 kg (range, 4 to 70 kg), respectively. All transplants were performed under tacrolimus and steroid immunosuppression. RESULTS: With a mean follow-up of 419 days (range, 5 to 795 days), patient and graft survival rates are 78% and 67%, respectively. One patient underwent a combined retransplantation with the standard technique 31 days after the primary allograft was destroyed by a native pancreatic fistula. Currently, all surviving recipients are at home and off TPN. DLSBTx allowed the expansion of the donor pool by transplanting 6 patients with donor to recipient weight ratio > or =1 and utilizing 2 less than 5-kg donors, including a neonatal donor. In 55% of the patients, chemical pancreatitis was observed during the early postoperative period. None of the duodenal allografts experienced signs of ischemia or leak. CONCLUSIONS: The technical advantages of this procedure include avoidance of a biliary reconstruction and simplification of the operative procedure. This, together with the feasibility of split or reduced liver grafting promises to increase the donor pool from neonates to adults. PMID- 10693684 TI - Pediatric recipients of three or more hepatic allografts: results and technical challenges. AB - BACKGROUND/PURPOSE: Children who require a liver transplant at an early age risk chronic allograft rejection (CAR) and other causes of allograft loss. Multiple retransplants may be required for long-term patient survival. The authors evaluate this approach based on our results and technical difficulties. METHODS: Charts of 7 children who received 3 or more liver transplants from 1989 to the present were reviewed retrospectively. RESULTS: A total of 151 children required liver transplantation at our institution since 1989. Of these, 4 boys and 3 girls (mean age, 6.2 years; range, 3 to 14 years) have received 3 or more allografts. The etiology of liver failure for the penultimate allograft was CAR (n = 6) and hepatic artery thrombosis (HAT; n = 1). Five cases required modification of portal vein or hepatic artery anastomoses. Two patients with vena caval strictures required supradiaphragmatic vena caval reconstruction. The original Roux-en-Y limb was adequate for biliary reconstruction in all cases. Five children currently are alive (survival rate, 71%) with good graft function having had a mean follow-up of 23 months (range, 2 to 48 mos.). CONCLUSIONS: The operative procedure for the multiple hepatic transplant child is challenging. The transplant team must be prepared for intraoperative issues such as extended organ ischemia time during hepatectomy, extensive blood loss, and potential need for creative organ revascularization techniques. Overall, multiple retransplant results are good and justify the use of multiple allografts. PMID- 10693685 TI - Extended left hepatectomy (left hepatic trisegmentectomy) in childhood. AB - BACKGROUND/PURPOSE: Extended left hepatectomy, also referred to as left hepatic trisegmentectomy, in which segments II, III, IV, V, and VIII are excised, is rarely performed in children. Experience with 7 such resections is reported to describe the anatomy, technique, indications, and outcomes of the operation. METHODS: The medical records of all pediatric patients treated at our institution over the last 15 years who underwent extended left hepatectomy were reviewed. Demographic information as well as operative, pathological, and follow-up data were analyzed. RESULTS: Seven patients underwent extended left hepatectomy over this period. There were 5 boys and 2 girls ranging in age between 4 months and 9 years with a median age of 3.1 years. Follow-up ranged from 8 months to 5 years with a median of 3.5 years. Diagnoses included hepatoblastoma (HB, n = 3), focal nodular hyperplasia (FNH, n = 1), leiomyosarcoma (LMS, n = 1), hepatocellularcarcinoma (HCC, n = 1), and metastatic neuroblastoma (NB, n = 1). All surgical margins were grossly negative. Median operative blood loss was 13 mL/kg (range, 5 to 32 mL/kg), and mean hospital stay was 9 days (range, 7 to 12 days). No major intra- or postoperative complications were encountered, and there was no perioperative mortality. The 3 HB patients, 1 FNH patient, 1 LMS patient, and 1 NB patient are without evidence of disease, whereas the 1 child with HCC died of recurrent and distant disease. The 6 surviving children have normal hepatic function. CONCLUSION: Although technically challenging and rarely performed, extended resection of the left hepatic lobe is feasible in children and can yield curative results with minimal morbidity. PMID- 10693686 TI - Aggressive surgery is unwarranted for biliary tract rhabdomyosarcoma. AB - BACKGROUND/PURPOSE: Rhabdomyosarcoma (RMS) of the biliary tract is rare, and, in addition to multiagent chemotherapy with or without radiotherapy (RT), some investigators recommend aggressive surgery. To assess the role of surgery, records of all 25 eligible patients with biliary RMS enrolled in IRSG studies I through IV from 1972 to 1998 were reviewed. METHODS: Treatment included surgery with or without vincristine, dactinomycin, cyclophosphamide, doxorubicin, cisplatin, etoposide, ifosfamide, and with or without RT. Data evaluated included clinical presentation, treatment, complications, and outcome. RESULTS: Diagnostic imaging identified the primary tumor but failed to identify regional metastases. Despite aggressive surgery, gross total resection at diagnosis was possible in only 6 cases, 2 of which had negative surgical margins. Although only 6 (29%) patients without distant metastases underwent gross total resection, estimated 5 year survival rate was 78% (95% CI 58%, 97%). Infectious complications were common and frequently associated with external biliary drains. Five (20%) died within the first 2 months, 3 of sepsis. CONCLUSIONS: Surgery is critical for establishing an accurate diagnosis and determining the extent of regional disease. Gross total resection is rarely possible despite aggressive surgery, and outcome is good despite residual disease after surgery. External biliary drains increase the risk of postoperative infectious complications. PMID- 10693687 TI - Preoperative staging, prognostic factors, and outcome for extremity rhabdomyosarcoma: a preliminary report from the Intergroup Rhabdomyosarcoma Study IV (1991-1997). AB - BACKGROUND: During the fourth Intergroup Rhabdomyosarcoma (RMS) Study (IRS IV, 1991-97), a preoperative staging system was evaluated prospectively for the first time. The authors evaluated this staging system and the role of surgery in extremity RMS in contemporary multimodal therapy. METHODS: A total of 139 patients (71 girls; median age, 6 years) were entered in IRS IV with extremity site RMS. Stage was assigned by the IRSG Preoperative Staging System. Postsurgical group was determined by tumor status after initial surgical intervention. Multivariate analysis was performed using all pretreatment factors that were significant by univariate analysis, including clinical Group (i.e., I through IV), tumor invasiveness (T1,T2), nodal status (N0,N1), and tumor size (< or > or =5 cm). Failure-free survival rates (FFS) and survival rates were estimated using the Kaplan and Meier method. RESULTS: Preoperative staging and clinical group distribution were as follows: Stage 2, n = 34; Stage 3, n = 73; Stage 4, n = 32; Group I, n = 31; Group II, n = 21; Group III, n = 54; Group IV, n = 33. Three-year FFS was 55%, and the overall survival rate was 70%. Eighty seven patients had either unresectable, gross residual disease (Group III) or metastases (Group IV). FFS was significantly worse for these patients with advanced disease, compared with that for patients with complete resection or with only microscopic residual tumor (i.e., Group I or II; Group I, 3-year FFS, 91%; Group II, 72%; Group III, 50%; Group IV, 23%; P<.001). Lymph nodes were evaluated surgically in 76 patients with positive results in 38. Clinically, 13 additional patients had nodal disease. Both stage and group were highly predictive of outcome and were highly correlated. By multivariate analysis, none of the other variables were predictors of FFS. CONCLUSIONS: This review confirms the utility of pretreatment staging for stratification of patients with extremity RMS with widely different risks of relapse, thereby paving the way for development of risk based therapy. Group (operative staging) remains the most important predictor of FFS, emphasizing the importance of complete gross resection at initial surgical intervention, when feasible without loss of limb function. The high incidence of nodal disease in the patients who had lymph node biopsy confirms the need for surgical evaluation of lymph nodes to ensure accurate staging in children with extremity rhabdomyosarcoma. PMID- 10693688 TI - Sonographic prognostic factors in fetuses with sacrococcygeal teratoma. AB - BACKGROUND: A subset of fetuses with sacrococcygeal teratoma (SCT) develops hydrops caused by high-output heart failure. Identification of fetuses at risk for hydrops is important because fetal intervention may reverse the pathophysiology of the disease. To date, no reliable sonographic prognostic factors have been identified. METHODS: An experienced sonologist reviewed the sonographic records of 17 fetuses with SCT referred to the authors' institution. Size of the tumor was measured and corrected for fetal size. The appearance of the tumor (solid versus cystic) and its vascularity were graded on a subjective scale of 1 to 5. RESULTS: Only 4 of 12 fetuses that had hydrops survived; of the survivors, 3 had undergone fetal intervention. All nonhydropic fetuses survived. Fetuses with hydrops had tumors that were mainly solid and highly vascular, whereas nonhydropic fetuses had predominantly cystic tumors with comparatively less vascularity. There was no significant difference in tumor size between these 2 groups. CONCLUSIONS: Fetuses with SCT that are mainly solid in appearance and are highly vascularized have a higher risk of getting hydrops in utero. Tumor size is not an independent prognostic factor. PMID- 10693689 TI - Testicular damage after surgical groin exploration for elective herniorrhaphy. AB - BACKGROUND/PURPOSE: Controversy exists whether to explore the contralateral groin in boys during unilateral herniorrhaphy. Proponents claim there is minimal risk of injury to the cord structures and developing testicle with contralateral exploration. However, findings have shown testicular atrophy occurred in 1% to 2% of patients after herniorrhaphy, and vasal damage is possible after routine manipulation of the spermatic cord. This study investigated the effect of routine surgical exploration of the prepubertal groin on testicular development and future fertility. METHODS: Twenty-four prepubertal Wistar rats were divided equally into 2 groups. Group 1 (sham) rats underwent unilateral inguinoscrotal incision only. Group 2 (experimental) rats underwent unilateral inguinoscrotal exploration with manipulation of the cord structures as in human inguinal exploration. At maturation, the fertility and fecundity of the males were assessed. After mating, testes were examined for mass, volume, mean seminiferous tubule diameter (MSTD), and mean testicular biopsy score (MTBS). The vasa were examined for histological injury and vasal diameter. Statistical comparisons were made by paired t test and Mann-Whitney rank sum test. RESULTS: There was a difference between the volumes of the testes when comparing the operative and nonoperative side of the 2 groups (experimental, deltavol = -0.063+/-0.123; sham, deltavol = +0.067+/-0.137; P = .029). There also was a trend toward a smaller testicular mass when comparing the two sides (experimental, deltamass = -0.045+/ 0.101; sham, Amass = +0.048+/-0.123; P = .057) but did not reach significance. The MSTD and MTBS were similar between the ipsilateral and contralateral testes in both groups. Likewise, the MSTD and MTBS were similar when comparing the 2 groups. All male rats in both groups were fertile. The number of offspring produced and the number of female rats impregnated were similar between the 2 groups. There was no histological evidence of vasal injury in any of the experimental spermatic cords. The vasal diameters were similar between the 2 groups. CONCLUSION: Surgical manipulation of the prepubertal spermatic cord imparts a small, but statistically significant morphological change in testicular size without a deleterious effect on testicular development, fertility, or fecundity. PMID- 10693690 TI - Alternative method for repair of the difficult infant hernia. AB - BACKGROUND: Inguinal hernia repair in the small infant is often technically difficult. An alternative operative approach has been developed that can simplify troublesome repairs, while decreasing the potential risks of damage to cord structures and of recurrence. METHODS: Thirteen small infants (weight 1400-3000 grams) underwent inguinal hernia repair using a technique of direct closure of the internal inguinal ring via a trans-hernial sac approach. Dissection of cord structures from the sac was avoided. RESULTS: All hernia repairs remained intact on follow-up of 4-28 months. One patient early in the series developed a noncommunicating hydrocele, prompting the addition of sac eversion to the original technique. CONCLUSIONS: An alternative method for simplified repair of difficult infant hernias has been used with success. While it does not supplant traditional repair technique for most patients, it should be considered for use in selected situations. PMID- 10693691 TI - Laparoscopic fundoplication in children with previous abdominal surgery. AB - BACKGROUND/PURPOSE: In our institution, many children requiring antireflux surgery for gastroesophageal reflux have had previous abdominal surgery, usually gastrostomy tube or ventriculoperitoneal (VP) shunt placement. The authors review their laparoscopic Nissen fundoplication (LNF) experience in children with previous abdominal surgery assessing surgical outcome. METHODS: A total of 82 consecutive LNFs performed at our institution between January 1996 and September 1998 were reviewed. Follow-up ranged from 1 month to 32 months (average, 8.9 months). LNF was performed without dividing short gastric vessels (Rosetti modification) through a standard 5-port technique. RESULTS: A total of 26 of 82 patients (31.7%) had previous abdominal surgery and were divided into 2 groups: gastrostomy (n = 17) and VP shunt (n = 11) with 2 crossovers. A total of 14 of 17 (82.3%) in the gastrostomy group had percutaneous endoscopic gastrostomy (PEG) placement versus 3 of 17 (17.6%) by open technique (open). Four patients in the VP group had multiple surgeries (range, 1 to 10, average, 2.3). LNF was completed in 25 of 26 (96.2%). One operation was converted to an open procedure because of severe adhesions. In 13 of 17 (76.5%) the previous gastrostomy was not taken down. In 4 of 17 (23.5%), the gastrostomy was taken down to complete the procedure: 2 of 3 (66.7%) of the open group versus 2 of 14 (14.3%) of the PEG group. All 11 (100%) of the VP group had successful LNF. Two of 11 (18.2%) had shunt dysfunction at 2 months (shunt infection) and 4 months (clogged distal shunt), respectively. There have been no cases of recurrent reflux, and all gastrostomies and VP shunts were functional at the time of this report. CONCLUSIONS: Previous abdominal surgery is common in children with gastroesophageal reflux disease requiring an antireflux procedure. The authors conclude from these preliminary results that laparoscopic Nissen fundoplication can be performed safely with minimal morbidity and excellent functional results in children with gastrostomies or ventriculoperitoneal shunts. PMID- 10693692 TI - Pyloromyotomy versus atropine sulfate for infantile hypertrophic pyloric stenosis. AB - PURPOSE: Atropine sulfate (atropine) and pyloromyotomy were compared for managing infantile hypertrophic pyloric stenosis (IHPS). METHODS: From 1996 to 1998, cases of IHPS treated surgically (pyloromyotomy; n = 20) or medically (atropine; n = 14) at separate institutions were compared retrospectively with regard to status on presentation, physical symptoms and signs, progress, and costs. Atropine was given orally, then intravenously if ineffective. Refractory cases were referred for pyloromyotomy. RESULTS: All subjects were matched for clinical and physiological status on admission. Oral atropine alone was effective in 11 cases, was converted to intravenous atropine in 2 cases, and was terminated in 1 case because of hematemesis. Two cases were referred for pyloromyotomy. All pyloromyotomies were successful. Atropine took on average, 2.6 days to take effect. The difference in time taken for normalization of pyloric muscle thickness between the 2 groups was not significant. Average time to return to full feeding was longer in the atropine group (P<.01). Costs were lower in the atropine group (P<.01). There were 2 wound infections in the pyloromyotomy group, but no adverse effects of atropine. There were no recurrences in either group. CONCLUSION: This study provides reasonable evidence to support a trial of atropine in IHPS. PMID- 10693693 TI - The modified Puestow procedure for complicated hereditary pancreatitis in children. AB - PURPOSE: The aim of this study was to evaluate the role of longitudinal pancreaticojejunostomy (modified Puestow procedure) in the treatment of complicated hereditary pancreatitis (HP) in children. METHODS: The authors reviewed their experience with the modified Puestow procedure for complicated HP in patients less than 18 years of age at a single tertiary care facility between 1973 and 1998. Main study outcomes included surgical morbidity and mortality, pre and postoperative pancreatic function, number of hospitalizations, and percentile ideal body weight (IBW). RESULTS: Twelve patients (6 boys and 6 girls) with a mean age of 9.3 years were identified. Presenting diagnoses were abdominal pain (n = 10), failure to thrive (n = 4), pancreatic pleural effusion (n = 2), and pancreatic ascites (n = 1). Blood loss was greater in patients who underwent distal pancreatectomy to localize the duct (n = 6) than in those who underwent direct transpancreatic duct localization (n = 6; 29.1+/-6.8 v. 8.3+/-3.7 mL/kg; P = .03). Other complications in patients who underwent distal pancreatectomy included splenic devascularization requiring splenectomy (n = 1) and postoperative intraabdominal bleeding with subsequent left subphrenic abscess (n = 1). There was no surgical mortality. Five patients had steatorrhea preoperatively that resolved in 4 patients postoperatively and was well controlled in the fifth. Mean number of hospitalizations for pancreatitis in the 5 years after surgery were markedly less than in the 5 years preceding surgery (0.4+/-0.2 v. 3.5+/-0.5; P = .01, n = 9). Percentile ideal body weight tended to increase within the first postoperative year (24.6+/-6.8 v. 45.0+/-8.3; P = .07, n = 9), and by the third year this trend was clearly significant (27.0+/-7.2 v. 60.9+/-9.5; P = .01, n = 8). CONCLUSIONS: In children with complicated HP, the modified Puestow procedure improves the quality of life by improving pancreatic function, decreasing hospitalizations, and increasing the percentile ideal body weight. Direct pancreatic duct localization during the procedure had a lower morbidity rate than localization via distal pancreatectomy. It is our impression that surgery performed in the early stage of complicated disease may preserve pancreatic function. PMID- 10693694 TI - T-tube ileostomy for meconium ileus: four decades of experience. AB - BACKGROUND/PURPOSE: The T-tube ileostomy was first used at Texas Children's Hospital in 1959. The purpose of this study is to update the experience since the initial report of this technique in 1981. METHODS: A database of 448 patients with cystic fibrosis (CF) seen in the authors' institution was used to identify 83 patients (18.5%) who presented with meconium ileus. The clinic and hospital charts of these patients were reviewed retrospectively to identify patients who had undergone placement of a T-tube ileostomy. RESULTS: Surgery was performed in 60 of 83 patients for complications of meconium ileus or failure to evacuate the meconium after a contrast enema. Of these patients, 21 of 60 (35%) underwent placement of a T-tube ileostomy. An additional 8 patients were identified who underwent placement of a T-tube ileostomy but were not included in the CF database, for a total of 29 patients who have been treated with T-tube ileostomy since 1959 at Texas Children's Hospital. Five patients were excluded from analysis because of insufficient data or misdiagnosis. One of the 24 patients in the series died of complications of prematurity. A total of 20 of 23 patients had resolution of their meconium ileus after T-tube irrigation with n-acetylcysteine or pancreatic enzymes. Three patients required additional surgery to relieve persistent bowel obstruction. All patients had the T-tube removed within the first 8 weeks after surgery. Two patients required subsequent repair of an incisional hernia. There were otherwise no complications of this procedure, with an average follow-up of 11.5 years. CONCLUSION: In patients with uncomplicated meconium ileus unrelieved by contrast enema, the T-tube ileostomy is an effective and safe treatment. PMID- 10693695 TI - A novel technique for correction of intestinal atresia at the ligament of Treitz. AB - PURPOSE: After reconstruction of jejunal atresias at the ligament of Treitz, many patients do not respond to simple tapering and anastomosis requiring repetitive operations because of dysfunction of dilated proximal bowel. A new operative approach using lateral duodenectomy and duodenojejunostomy (LDAD) is reported. METHODS: Three infants with atresias within 10 cm of the ligament of Treitz were treated with LDAD, and their records are reviewed retrospectively. The entire duodenum is visualized after creating a malrotation; this is followed by opening the dilated duodenum and resecting dilated proximal jejunum. The resection is extended proximally, incorporating the lateral duodenal incision, excising the lateral duodenal wall, and preserving the ampulla. The residual duodenum is fashioned into a tube and anastomosed to the spatulated distal jejunum. RESULTS: Three infants underwent this procedure over a 4-year period. Two had undergone tapering enteroplasties previously but were unable to tolerate oral feedings; 1 infant had LDAD primarily. All were ultimately successfully managed by LDAD and were feeding within 14 days. Follow-up is from 14 to 49 months. CONCLUSION: Although experience is limited to 3 patients, the prompt return of intestinal function with LDAD may justify primary use of this more radical procedure in difficult-to-treat proximal atresias. PMID- 10693696 TI - Glucagonlike peptide-2 analogue enhances intestinal mucosal mass after ischemia and reperfusion. AB - BACKGROUND/PURPOSE: Glucagonlike peptide-2 (GLP-2), a product of the posttranslational processing of proglucagon, has been shown to enhance mucosal mass and function in both normal intestine and in the residual intestine after massive small bowel resection. This study was designed to determine if a synthetic, protease-resistant analogue of GLP-2 (GLP-2alpha) can enhance mucosal mass in small intestine after ischemia and reperfusion (I/R) injury. METHODS: Ten young adult male Sprague-Dawley rats underwent laparotomy and superior mesenteric artery occlusion for a period of 40 minutes. During this period of ischemia, each rat underwent placement of a jugular venous catheter that was connected to a subcutaneously placed osmotic pump designed to deliver its contents over 3 days. The rats were divided into 2 groups based on the contents of the pumps: group 1, saline at 1 microL/h (n = 6) and group 2, GLP-2alpha at 100 microg/kg/d (n = 4). Three days after insertion of the pumps the small intestine was harvested from the surviving rats for determination of mucosal DNA and protein content. Statistical analysis was performed using unpaired Student's t test. RESULTS: After I/R injury to the small intestine, a 3-day systemic infusion of GLP-2alpha significantly increased mucosal DNA content 41% (P<.05) and mucosal protein content 60% (P<.05) when compared with saline-treated controls. In addition, infusion of GLP-2alpha reduced mortality from 50% to 25%. CONCLUSIONS: These data show for the first time that GLP-2alpha enhances mucosal mass following I/R injury to the small intestine. GLP-2alpha may be of benefit to patients with intestinal ischemia syndromes such as necrotizing enterocolitis and midgut volvulus. PMID- 10693697 TI - Hormonal therapy for short bowel syndrome. AB - PURPOSE: Treatment of short bowel syndrome (SBS) can be difficult; this study examines the effect of parental administration of different peptide hormones in a rat model of SBS. METHODS: Juvenile male Lewis rats (220 to 240 g) underwent resection of the proximal 90% of small bowel and were assigned randomly to treatment groups: growth hormone (GH), insulinlike growth factor-1 (IGF-1), glucagonlike peptide-2 (GLP-2; given as ALX-0600, a potent protease resistant analogue of the human GLP-2), control-resected (Con-R), or control-transected (Con-T). Drugs were delivered by continuous subcutaneous infusion via Alzet mini pumps: controls received equivalent volumes of drug vehicle. Animals were pair fed (23 g chow per day) and followed up for 14 days monitoring weight gain. Animals were killed and active transport, hormone profiles, and intestinal morphology were assessed. RESULTS: Hormonal treatments significantly increased weight gain in all groups (GH, 9.9+/-4.9; IGF-1, 6.0+/-9.6; and GLP-2, 0.8+/-2.7 v. -6.2+/-4.7 in untreated resected animals [weight as percentile initial weight]). This was associated with a significant alteration in intestinal morphology in the IGF-1-treated animals, and an increase in glucose transport rates in all hormonally treated animals when compared with untreated control resected animals. CONCLUSIONS: These results show that IGF-1, GH, and GLP-2 all improve short-term weight gain after massive bowel resection in a rat model. The effects seen on weight gain may be caused by improved dietary nutrient absorption from an increase in the intestinal surface area or increase in transporting activity or alterations in the metabolic efficiency of the animal. These findings suggest further studies of these therapies as treatment for short-bowel syndrome are indicated. PMID- 10693698 TI - Intestinal adaptation occurs independent of transforming growth factor-alpha. AB - BACKGROUND/PURPOSE: Signal transduction via the epidermal growth factor receptor (EGFR) is critical for intestinal adaptation after massive small bowel resection (SBR). Although it has been assumed that the major ligand for the EGFR during adaptation is EGF, the role for transforming growth factor-alpha (TGF-alpha), another major ligand for the EGFR is unknown. The purpose of this study was to test the hypothesis that TGF-alpha is an important ligand for the EGFR during intestinal adaptation. METHODS: Wild-type mice (C57BI/6) underwent a 50% proximal SBR or sham operation (bowel transection or reanastomosis) and were then assigned randomly to receive either intraperitoneal TGF-alpha or placebo. In a separate experiment, SBR or sham operations were performed in mice lacking TGF-alpha (Waved-1). After 3 days, adaptation was measured in the ileum. RESULTS: Exogenous TGF-alpha enhanced intestinal adaptation in the wild-type mice after SBR as shown by increased ileal wet weight and DNA content. Normal adaptation occurred in the mice lacking TGF-alpha as shown by increased ileal wet weight, protein and DNA content, proliferation, villus height, and crypt depth. CONCLUSIONS: Although exogenous TGF-alpha enhanced adaptation after massive SBR, adaptation was preserved in TGF-alpha-absent mice. These results refute TGF-alpha as an essential ligand for EGFR signaling during intestinal adaptation. PMID- 10693699 TI - Interleukin-11 enhances small intestine absorptive function and mucosal mass after intestinal adaptation. AB - BACKGROUND/PURPOSE: Interleukin-11 (IL-11) recently has been shown to enhance mucosal mass after massive small bowel resection (MSBR). However, enhanced mucosal mass may not correlate with increased substrate absorption. This study was designed to examine the effect of systemic administration of increasing doses of IL-11 on small intestine absorptive function and mucosal mass after MSBR. METHODS: Twenty-five Sprague-Dawley rats underwent an 80% small bowel resection and end-to-end jejunoileal anastomosis. Seven days after resection, all rats had placement of a jugular venous catheter connected to a subcutaneously placed osmotic pump. The rats were divided into 5 groups based on the content of the pump: group 1 (control, n = 5) received 0.1% bovine serum albumin (BSA) and groups 2 through 5 (n = 5 each) received IL-11 at 250, 500, 750, and 1,000 microg/kg/d, respectively. After a 14-day infusion period, [14C] galactose and [14C] glycine absorption was measured using an in vivo closed-recirculation technique. Mucosal DNA content also was determined for each group. Statistical analysis was performed by analysis of variance and expressed as mean +/-SEM. RESULTS: IL-11 administered at 250 microg/kg/d, a dose used in previous studies, did not significantly affect substrate absorption. However, compared with the control group, administration of higher doses of IL-11 produced a significant increase in substrate absorption and mucosal mass. The dose of IL-11 producing the overall optimal response based on the parameters measured (galactose absorption, 72% increase over control; glycine absorption, 112% increase over control; and DNA content, 98% increase over control) was 750 microg/kg/d. CONCLUSIONS: In addition to an increase in mucosal mass, these data show for the first time that IL-11 enhances absorptive function beyond the normal adaptive response after MSBR. Furthermore, the maximum effect of IL-11 on absorptive function was shown at 750 microg/kg/d, which is 3 times the dose used in previously reported studies. This study suggests that IL-11 may be useful clinically in patients with inadequate intestinal function. PMID- 10693700 TI - Overexpression of Fas-ligand by neuroblastoma: a novel mechanism of tumor-cell killing. AB - BACKGROUND/PURPOSE: Binding of Fas ligand (Fas-L) to the membrane-bound Fas receptor incites a series of intracellular events that results in programmed cell death or apoptosis. Although this apoptotic phenomenon plays a key role in down regulating cytotoxic T cells, the authors have shown previously that pancreatic beta cells (bTC) overexpressing Fas-L paradoxically undergo accelerated rejection that is dependent on a Fas/Fas-L interaction. This study evaluates whether a neuroblastoma (NB) cell line manipulated to overexpress Fas-L undergoes similar destruction and whether tumor-specific protective immunity can be produced. METHODS: The authors transfected NB cells (SK-N-MC) with either mFas-L cloned into a pcDNA3.1/Zeo plasmid vector (NB/Fas-L) or with the vector alone (NB/control). Successful transfection of Fas-L was characterized by reverse transcription polymerase chain reaction (RT-PCR) and the ability of transfectants to induce apoptosis of Fas-sensitive T cells (Jurkat). Expression of Fas and Fas L in untransfected NB clones was characterized by immunohistochemistry and RNase protection assay (RPA). Apoptosis was measured by FACScan analysis using an Annexin V assay. A total of 3x10(6) NB/control and NB/Fas-L cells were implanted subcutaneously into the hind leg of Balb/C SCID mice. Tumor-specific protective immunity was also tested in this model by inoculating mice with NB/Fas-L before implanting NB/control cells. RESULTS: Zeocin resistance and RT-PCR confirmed successful transfection of Fas-L into NB cells. Fas Ligand transfectants induced apoptosis in 17.6%+/-2.9% of Fas-sensitive T cells, whereas controls induced apoptosis in only 2.8%+/-1.2% (P = .01, n = 3). Although Fas appears to be constitutively expressed by NB in low amounts, introduction of Fas-L into NB cells did not induce suicide or affect tumor cell growth in vitro. In vivo, NB cells expressing Fas-L failed to grow in SCID mice (n = 3), whereas controls grew rapidly in all animals until death (n = 3). NB/control cells implanted into the opposite leg of mice that rejected initial NB/Fas-L transfectants also grew rapidly (n = 3) implying no protective immunity. CONCLUSIONS: Overexpression of Fas-L in NB clones targets such cells for rapid destruction even in immune compromised hosts, suggesting potential utility of Fas-L in combating NB. In this SCID mouse model, the observed effect is probably neutrophil mediated and does not provide tumor-specific protective immunity. PMID- 10693701 TI - Coordinated interdisciplinary management of pediatric intestinal failure: a 2 year review. AB - BACKGROUND/PURPOSE: Intestinal failure is a complex metabolic process that results from malabsorption and malnutrition and provides challenges for a variety of pediatric subspecialists. The purpose of this study was to evaluate the effect of coordinated interdisciplinary team management of children with intestinal failure on nutritional outcome measures. METHODS: The Intestinal Care Center (ICC) is staffed with an interdisciplinary team of pediatric specialists including a gastroenterologist, pediatric surgeon, transplant surgeon, clinical dietitians, and a nutrition support nurse. Using an established registry, the authors conducted a comprehensive evaluation of patient data including anthropometric measures, organ system function, and mode of nutrition support. Disease-associated complications including micronutrient deficiencies, growth delay, and death also were monitored. Nutritional outcome was assessed by transition from enteral to oral feeding, cessation of total parenteral nutrition (TPN), and maintenance of linear growth. RESULTS: Since the inception of the ICC in 1996, 103 patients (69 boys, 34 girls) with intestinal failure have been evaluated with a median age of 2.6 years (range, 0.2 to 21.3 years). Mode of nutritional therapy on initial consultation included TPN (n = 76, 74%), enteral feedings (n = 6, 6%) and oral intake (n = 21, 20%). After intensive management of the 76 patients who were TPN dependent, 22 (29%) subsequently have been weaned from TPN (duration, 0.2 to 17.5 years) to oral (n = 14), oral-enteral (n = 4) or enteral feedings (n = 4). Of the 6 patients who were receiving enteral feedings, 4 (67%) were transitioned to oral feedings. Sixty-eight patients (66%) had evidence of hepatic disease. Of these, 10 underwent transplant, and 23 died (2 posttransplant). Linear growth velocity of neither pre- nor postpubescent patients significantly improved during the 2-year study period. CONCLUSION: Data registry establishment and concurrent interdisciplinary team management of children with intestinal failure provides for innovative treatment approaches and a foundation for retrospective or prospective assessment of children with disease. PMID- 10693702 TI - Radiofrequency thermal ablation: a potential treatment for hydropic fetuses with a large chest mass. AB - BACKGROUND/PURPOSE: Physiological changes secondary to a rapidly enlarging fetal chest mass can lead to nonimmune hydrops, which is a predictor of impending fetal demise. Currently, open fetal surgery is offered for specific patients with hydrops before 32 weeks' gestation. The authors asked if a less-invasive technique, radiofrequency thermal ablation (RTA), could be applied safely to the destruction of fetal lung tissue. METHODS: Time-dated pregnant ewes at 120 to 125 days' gestation (term, 140 to 145 days) underwent RTA through a hysterotomy (n = 3) or in a transuterine fashion under ultrasound guidance (n = 4). The probe is a 15-gauge needle with a maximal 2-cm deployment sphere at its tip. By varying the intensity and duration of treatment, the power settings were optimized to create a defined area of ablation with minimal surrounding tissue injury. Five of the 6 fetal lambs were killed acutely, and 1 was followed up for 1 week with frequent ultrasound examinations. RESULTS: Gross examination of the animals killed acutely showed a consistent area of ablation using 10 watts of applied power for 3 minutes. Direct coagulation necrosis lesions ranged from 1x1 cm to 2x2 cm in size depending on the extent of the probe deployment. One animal was followed up for 1 week and showed no major adverse physiological effects. CONCLUSION: Radiofrequency thermal ablation can be performed safely in this fetal sheep model to create a controlled area of lung tissue ablation. PMID- 10693703 TI - Differential apoptosis gene expression in pediatric tumors of the kidney. AB - BACKGROUND/PURPOSE: Apoptosis, or programmed cell death, is essential in maintaining normal homeostasis of tissues. The process of apoptosis is controlled by numerous pro- and antiapoptotic factors. Variations in expression of such factors may account for some variations in tumor behavior. This study evaluates the expression of apoptotic mRNA species in pediatric renal tumors to determine whether a pattern of differential apoptosis gene expression correlates with tumor grade and type. METHODS: Twenty-five frozen tissue specimens were obtained from patients undergoing biopsy or resection of pediatric renal tumors before chemotherapy: Wilms' tumor stage II (WT-II, n = 4); Wilms' tumor stage III/IV (WT III/IV, n = 4); clear cell sarcoma of the kidney stage III (CCSK, n = 2); rhabdoid tumor of the kidney stage III/IV (RTK, n = 4); and normal kidney (NK, n = 11). An RNase Protection Assay (RPA) was performed for 19 pro- and antiapoptotic mRNA species to detect and quantify expression (percentage of GAPDH expressed). Expression of specific mRNAs of interest were confirmed by Western Blot (WB). RESULTS: The expression of apoptotic mRNA species varied markedly between tumors. WT-II expressed greater amounts of proapoptotic receptor mRNA than CCSK or RTK. (Fas, 17.0+/-2.7% v. 2.5+/-0.5% v. 3.3+/-0.9%; P<.02; DR5, 77.0+/-8.8% v. 13.5+/-0.5% v. 27.0+/-4.8; P<.001; TNF-R, 71.3+/-17.0% v. 21.0+/ 4.0% v. 29.0+/-5.0%; P<.07, respectively). Surprisingly, antiapoptotic factors (e.g., bcl-2 and bcl-xl) were not overexpressed in poor prognostic tumors (CCSK, RTK) compared with those with good prognosis (WT). Expression of TRAIL (a ligand for DR4 and DR5) was significantly lower in CCSK and RTK than in normal kidney (9.5+/-1.5% v. 56.1+/-10.1%; P = .01). CONCLUSIONS: Proapoptotic receptors are expressed at greater levels in good prognostic tumors, and this finding is compatible with their clinical behavior. Knowledge of differential apoptotic gene expression is of potential value in predicting prognosis and treating such tumors with targeted ligands. PMID- 10693704 TI - Pancreatitis caused by duodenal duplication. PMID- 10693705 TI - Outcome analysis for gastroschisis. PMID- 10693706 TI - Prolonged intestinal exposure to amniotic fluid does not result in peel formation in gastroschisis. PMID- 10693707 TI - Quality improvement for diagnostic neuroangiography and other procedures. PMID- 10693708 TI - Animal models for atherosclerosis, restenosis, and endovascular graft research. AB - Animal models have significantly advanced our understanding of the mechanisms of atherosclerosis and restenosis formation and the evaluation of therapeutic options. The current focus of research is on preventive strategies against restenosis and includes pharmacologic and biologic interventions directed primarily against smooth muscle cell proliferation, endovascular devices for recanalization and/or drug delivery, and an integrated approach using both devices and pharmacobiologic agents. Devices aimed at the percutaneous endoluminal exclusion of aortic aneurysms have also generated interest recently. The experience over many decades with animal models in vascular research has established that a single, ideal, naturally available model for atherosclerosis, restenosis, or for that matter aneurysm formation, does not exist. Presently, rabbits and pigs are favored for the former two areas of study, and dogs and sheep appear to provide suitable models for testing devices for endoluminal repair of aneurysms. The development of transgenic variants of currently available models may widen our options in the future. Nevertheless, an appreciation of the individual features of natural or stimulated disease in each species is of the utmost importance for the proper design and execution of relevant experiments. PMID- 10693709 TI - Regulatory and legal implications of modifying FDA-approved medical devices. PMID- 10693710 TI - Management of patient skin dose in fluoroscopically guided interventional procedures. AB - PURPOSE: To simulate dose to the skin of a large patient for various operational fluoroscopic conditions and to delineate how to adjust operational conditions to maintain skin dose at acceptable levels. MATERIALS AND METHODS: Patient entrance skin dose was estimated from measurement of entrance air kerma (dose to air) to a 280-mm water phantom for two angiographic fluoroscopes. Effects on dose for changes in machine floor kVp, source-to-skin distance, air gap, electronic magnification, fluoroscopic dose rate control settings, and fluorographic dose control settings were examined. RESULTS: Incremental changes in operational parameters are multiplicative and markedly affect total dose delivered to a patient's skin. For long procedures, differences in doses of 8 Gy or more are possible for some combinations of operational techniques. CONCLUSIONS: Effects on skin dose from changes in operational parameters are multiplicative, not additive. Doses in excess of known thresholds for injury can be exceeded under some operating conditions. Adjusting operational parameters appropriately will markedly reduce dose to a patient's skin. Above all other operational factors, variable pulsed fluoroscopy has the greatest potential for maintaining radiation exposure at low levels. PMID- 10693711 TI - Lower extremity arteriography with use of iodinated contrast material or gadodiamide to supplement CO2 angiography in patients with renal insufficiency. AB - PURPOSE: To determine if the use of nonionic contrast material, as compared to the use of gadodiamide to supplement carbon dioxide angiography in patients with peripheral vascular disease (PVD) and chronic renal insufficiency (CRI), results in significant worsening of renal function. MATERIALS AND METHODS: Lower extremity angiographic procedures (diagnostic and diagnostic/intervention) were performed in 40 patients with CRI (baseline serum creatinine [Cr] > 1.5 mg/dL) using CO2 alone or CO2 supplemented with the use of either nonionic contrast material or gadodiamide (up to 0.4 mmol/kg). Serum creatinine levels were obtained before the procedure and at 48 hours after the procedure. The peak Cr level was also determined for patients with a significant (> 0.5 mg/dL) Cr elevation. RESULTS: Forty-two lower extremity angiographic procedures (19 diagnostic and 23 diagnostic/interventions) were performed in 40 consecutive patients from August 1997 to October 1998, with a mean preprocedure Cr of 2.2 mg/dL and a mean postprocedure Cr of 2.4 mg/dL. Twenty-five of the 40 patients (63%) had diabetes mellitus. Fifteen procedures, including six interventions, were performed utilizing CO2 and nonionic contrast material in 15 patients. Six of these 15 patients (40%) demonstrated a Cr increase > 0.5 mg/dL at 48 hours. Seven procedures, including two interventions, were performed with CO2 alone in seven patients. No patients in this group demonstrated an increase in serum creatinine of greater than 0.5 mg/dL at 48 hours. Twenty procedures, including 15 interventions, were performed with CO2 and gadodiamide in 18 patients. In one of these 20 procedures (5%) there was an increase in Cr > 0.5 mg/dL at 48 hours The difference in worsening renal function for the nonionic contrast group (six of 15) compared with the CO2/gadodiamide group (one of 20) was statistically significant (P = .03). When comparing the use of CO2 and nonionic contrast material versus CO2 alone and with gadodiamide (six of 15 versus one of 27), the difference is also statistically significant (P < .01). The average volume of supplemental contrast material was similar in the nonionic contrast material and gadodiamide groups, as was the average volume of supplemental nonionic contrast material in the six patients with an increased Cr. CONCLUSION: The use of small volumes of nonionic contrast material to supplement CO2 angiography in patients with PVD and CRI can be associated with a significant increased risk of worsening renal function when compared to angiography performed with CO2 alone or CO2 and gadodiamide. PMID- 10693712 TI - Safety of short stay observation after peripheral vascular intervention. AB - PURPOSE: To determine whether short observation periods (less than or equal to 4 hours) are safe in outpatients undergoing arterial peripheral vascular interventions. MATERIALS AND METHODS: A retrospective review of 203 patient medical records from the Interventional Vascular Department for 239 lower extremity or abdominal procedures (161 men and 78 women) during a 5-year period was completed. The average patient age was 62.2 years (range, 32-83 years). Thirty-six patients had more than one procedure. Indication, intervention, coagulation status, complication rate, and hospitalizations within 7 days after discharge from the short stay unit (SSU) were reviewed and the outcome was measured. Patients were grouped according to the length of their observation period (< or =4 hours or >4 hours) for statistical analysis. RESULTS: In 85% of the procedures (204 procedures), claudication was the primary indication for intervention. Angioplasty (203 procedures) was also commonly performed. Ninety procedures (38%) required stent placement, and other interventional procedures performed were pulse-spray thrombolysis (eight procedures), atherectomy (two procedures), and stent-graft placement (one procedure). None of the patients required hospitalization as a result of their radiologic intervention within 7 days after discharge from the SSU. Specifically, there were no major "at home" complications in patients discharged after an observation period of < or =4 hours. Two patients were admitted for outpatient procedures and were subsequently hospitalized as a result of a complication from the procedure. The complication rate (including minor complications) was 8% (seven of 87) in the < or =4 hour observation period group compared with 24.3% (37 of 152) in the >4 hour group (P < .01). This difference was due to a greater number of minor hematomas in the >4 hour group. CONCLUSION: Based on the authors' findings, many interventional vascular procedures can be performed safely on an outpatient basis with relatively short observation times. Early discharge from the SSU did not result in an increased readmission rate to the hospital because of delayed complications. PMID- 10693713 TI - Exercise-induced external iliac artery intimal fibrosis: confirmation with intravascular ultrasound. PMID- 10693714 TI - Retrograde catheterization of the inferior mesenteric artery to treat endoleaks: anatomic and technical considerations. PMID- 10693715 TI - Use of transjugular intrahepatic portosystemic shunts during lytic therapy of extensive portal splenic and mesenteric venous thrombosis: long-term follow-up. PMID- 10693716 TI - Transcatheter arterial embolization of ruptured pseudoaneurysms with coils and n butyl cyanoacrylate. AB - PURPOSE: To evaluate the clinical efficacy of transcatheter arterial embolization with n-butyl cyanoacrylate (NBCA) for ruptured pseudoaneurysms, which are difficult to control by coil embolization alone. MATERIALS AND METHODS: Ruptured pseudoaneurysms developed at the celiac trunk (n = 1), gastroduodenal artery (n = 2), pancreatic arcade (n = 1), hepatic artery (n = 3), renal artery (n = 1), and intercostal artery (n = 1) in nine patients. NBCA was mixed with iodized-oil (1:2) and injected via the 3-F microcatheter under fluoroscopic guidance, after the catheter was advanced close to the pseudoaneurysm. Coil embolization was performed to control blood flow before administration of NBCA in seven patients. NBCA was injected immediately after coil embolization in four patients. Embolization with NBCA was performed for recurrent bleeding that occurred within 1-21 days (mean, 10.7 days) after initial coil embolization in three patients. Two patients with peripheral pseudoaneurysms underwent embolization with NBCA alone. RESULTS: The NBCA mixture was visible under fluoroscopy, and was useful in monitoring the embolization process and deciding the endpoint. Embolization was technically successful without major complications in all patients. Pseudoaneurysms and afferent and efferent arteries were eliminated immediately after embolization. Bleeding was stopped after embolization in all cases. Rebleeding did not occur in any patient during their follow-up periods of 0.7 69.5 months (mean, 17.9 months). CONCLUSION: Embolization with NBCA is a feasible and useful treatment for ruptured pseudoaneurysms, which are difficult to control by coil embolization alone. PMID- 10693717 TI - Gunther Tulip filter: preliminary clinical experience with retrieval. AB - PURPOSE: The Gunther Tulip filter is a permanent filter that has a hook to permit retrieval. The authors report their preliminary clinical evaluation of the filter with regard to feasibility and safety of retrieval. MATERIALS AND METHODS: Nine men and six women who ranged in age from 17 to 79 years (mean, 51 years) underwent treatment with use of the Gunther Tulip filter. Patients judged to require caval interruption for < 14 days were selected to receive the filter and retrieval was planned for all patients. Indications for filter placement were: recent pulmonary embolism (PE) or proximal deep vein thrombosis (DVT) with a contraindication to anticoagulation (11 patients), massive PE treated with thrombolytic therapy (one patient), PE with heparin-induced thrombocytopenia (one patient), and prophylaxis after major trauma (two patients). Patients were followed for inferior vena cava (IVC) thrombosis, bleeding, and recurrent DVT or PE. RESULTS: In all nine patients in whom it was attempted, the filter was successfully snared and retrieved via a jugular approach. The mean implantation period was 8.6 days (range, 5-13 days). Retrieval required 2.2-13 minutes (mean 5.3 minutes) of fluoroscopy. No caval injuries occurred as a result of retrieval. All retrieved filters had strands of organized thrombus on the filter struts. The patients were followed for 52-285 days (mean, 115 days) after retrieval. One patient developed a recurrent DVT 230 days after retrieval. No other patients developed a recurrent DVT and no patients developed IVC thrombosis, bleeding, or PE. Six filters were not retrieved: five because of an ongoing contraindication to anticoagulation and one because the patient died of causes unrelated to the filter. CONCLUSION: This preliminary study confirms the feasibility and safety of retrieval of the Gunther Tulip filter up to 13 days after implantation. PMID- 10693718 TI - Current practice of temporary vena cava filter insertion: a multicenter registry. AB - PURPOSE: To evaluate the current practice of temporary vena cava filter placement and its complications. MATERIALS AND METHODS: A multicenter registry was conducted from May 1995 until May 1997 using a standardized questionnaire. One hundred eighty-eight patients were evaluated. Patient characteristics, filter indications, filter characteristics, and complications were registered. RESULTS: Deep vein thrombosis was proven in 95.2% of the patients. Main filter indication was thrombolysis therapy (53.1%). Average filter time was 5.4 days. An Antheor filter was inserted in 56.4%, a Guenther filter in 26.6%, and a Prolyser filter in 17.%. Transfemoral filter implantation was slightly preferred (54.8%). Four patients died of pulmonary embolism (PE) during filter protection. Major filter problems were filter thrombosis (16%) and filter dislocation (4.8%). When thrombus was found in or at the filter before explantation, additional thrombolysis was performed in 16.7%, additional filter implantation in 10%, and thrombus aspiration in 6.7%; 4.8% of filters were replaced with permanent filters. DISCUSSION: Temporary vena cava filters are placed to prevent PE in a defined patient population. Despite their presence, PEs still occur in a small percentage. Problems of filter thrombosis and dislocation have to be solved. CONCLUSION: The results of this multicenter registry support the need for innovative filter design, as well as a randomized, prospective study. PMID- 10693719 TI - Balloon dilation and endobronchial stent placement for bronchial strictures after lung transplantation. AB - PURPOSE: To evaluate the effect of balloon dilation and endobronchial stent placement for bronchial fibrous stenoses and bronchomalacia after lung transplantation. MATERIALS AND METHODS: Bronchial dilation and/or stent placement was performed on 25 lung transplant recipients. Indications included severe dyspnea with postobstructive pneumonia (n = 24) and respiratory failure (n = 1). All patients underwent pulmonary function testing (PFT) before and after bronchial dilation, the results of which were evaluated for changes. A total of 63 procedures were performed between February 1996 and December 1998. Thirty-five lesions were treated (18 were due to bronchomalacia, 17 were due to fibrosis). Areas treated included the left mainstem bronchus (n = 11), bronchus intermedius (n = 10), right mainstem bronchus (n = 7), left upper lobe bronchus (n = 4), right lower lobe bronchus (n = 2), and right middle lobe bronchus (n = 1). Bronchoscopic and/or bronchographic follow-up ranged from 1 to 34 months (mean, 15 months). RESULTS: Six-month primary patency of stents placed for bronchomalacia was 71% (10 of 14), with three of the four occlusions caused by mechanical failure of Palmaz stents in the mainstem bronchi. Six-month primary patency for treatment of fibrous strictures was 29%. Secondary patency at 1 year was 100% for both bronchomalacia and fibrous strictures. After treatment, there was a significant improvement in mean PFT results (P = .01-.0001). There was one acute complication, obstruction of the left lower lobe bronchus by a Wallstent treated by dilating a hole in the side of the stent. CONCLUSIONS: Balloon dilation and stent placement are safe and effective for bronchial strictures and bronchomalacia after lung transplantation, resulting in significant improvement in PFT results. However, there is almost universal restenosis in patients treated for fibrous strictures necessitating reintervention for prolonged patency. PMID- 10693720 TI - US and fluoroscopic-guided percutaneous jejunostomy: experience in 49 patients. AB - PURPOSE: To assess the feasibility and safety of a variety of techniques for percutaneous jejunostomy. MATERIALS AND METHODS: Percutaneous jejunostomy was attempted on 53 occasions in 49 patients under US and fluoroscopic guidance. During the study period, thicker needles, Cope anchors, and intravenous glucagon were introduced to improve access, dilate, and immobilize the jejunum. Technical success rates, complications, catheterization period, and reasons for catheter removal were prospectively studied. Prognostic factors for successful procedures and complications were determined. RESULTS: Forty-six (87%) procedures were technically successful. Previous gastrointestinal surgery (P = .01) and a combination of thicker needles, Cope anchors, and intravenous glucagon (P = .0001) were associated with a higher technical success rate. Complications (n = 6; pericatheter leakage in four of six) were observed more frequently in older patients (P = .01). The 30-day mortality rate was 17%, one death may have been procedure related. Forty-three catheters were removed (elective, n = 36; other, n = 7) after 1-597 days (median, 49). Three catheters remained in situ for 139-482 days (median, 410). CONCLUSIONS: Percutaneous jejunostomy is a feasible and relatively safe technique for long-term feeding. Leakage is the main problem, which warrants additional study. PMID- 10693721 TI - Percutaneous diagnosis and treatment of biliary candidiasis. PMID- 10693722 TI - Autologous vein stent-graft: feasibility study. AB - PURPOSE: To evaluate expandable stents healed into vein wall as autologous vein stent-grafts for endoluminal grafting. MATERIALS AND METHODS: Balloon expandable stents were placed into external jugular veins of eight dogs. Stent and vein patency was followed by ultrasonography. Five weeks after stent placement, jugular veins with endothelialized stent were harvested. The autologous vein stent-grafts were divided into two groups. In group A, autologous vein stent grafts (n = 3) were placed immediately into Baker solution for microscopic examination. In group B, autologous vein stent-grafts (n = 3) underwent mechanical manipulation; they were compressed, mounted on angioplasty balloon, pushed through a 9-F sheath and dilated. The autologous vein stent-graft endothelialization and changes after mechanical manipulation were evaluated by light and electron microscopy. RESULTS: Stent placement was successful in seven dogs. One stent migrated into the pulmonary artery. One well placed stent was damaged by compression dressing and thrombosed. At 5 weeks, gross and microscopic examinations revealed the autologous vein stent-grafts were fully covered by a 0.115- +/- 0.036-mm-thick neointimal layer. Small wall thrombus was observed in one autologous vein stent-graft. Repeated manipulations did not result in any intimal damage or stent loosening in the autologous vein stent-grafts. CONCLUSION: Expandable stents healed into a vein have potential to be used as autologous vein stent-grafts for endoluminal grafting without risk of disruption during percutaneous transcatheter introduction. PMID- 10693723 TI - Local delivery of nadroparin via hydrogel-coated angioplasty balloon: effect on platelet deposition and smooth muscle cell proliferation--an experimental study. AB - PURPOSE: To assess the feasibility of intravascular delivery of nadroparin, a low molecular-weight heparin, via hydrogel-coated angioplasty balloons, and the effects of nadroparin delivered in this manner on platelet deposition and smooth muscle cell (SMC) proliferation. MATERIALS AND METHODS: Tritiated nadroparin was used to determine the nadroparin-carrying capacity of the hydrogel-coating, kinetics of release from the balloons, and, in four pigs, delivery of the nadroparin to the iliac arterial wall. Platelet deposition in nadroparin-treated iliac arteries versus contralateral iliac arteries dilated with saline-loaded, hydrogel-coated balloons was quantified in seven pigs using 111Indium-labeled platelets. Smooth muscle cell proliferation in nadroparin and saline-treated iliac arteries in 10 pigs was evaluated 7 days after angioplasty with use of proliferating cell nuclear antigen. RESULTS: Approximately 98 international units of nadroparin were delivered by the hydrogel-coated balloon, the majority to the angioplasty site and distal vessel. There was a trend toward decreased platelet deposition in nadroparin-treated arteries, but statistical significance was not achieved (P = .1563). Medial SMC proliferation was decreased in the nadroparin treated arteries in nine of 10 pigs (P = .0137). CONCLUSIONS: Hydrogel-coated balloons may be used to deliver nadroparin to the arterial wall, with measurable levels of the drug delivered to the site of angioplasty, and with resultant decrease in SMC proliferation. PMID- 10693724 TI - TIPS reduces hepatic asialoglycoprotein receptor concentration in healthy pigs. AB - PURPOSE: Technetium-99m-labeled diethylenetriamine pentaacetic acid galactosyl human serum albumin (TcGSA), a new agent for liver scintigraphy, is selectively attached to asialoglycoprotein receptors on liver cell membranes. A direct correlation exists between asialoglycoprotein receptor concentration, [R]o and hepatic functional reserve. The purpose of this study was to determine the effects of transjugular intrahepatic portosystemic shunting (TIPS) on hepatic asialoglycoprotein receptor concentration in pigs without liver parenchymal disease. MATERIALS AND METHODS: TIPS was performed in eight pigs with use of a single 10-mm-wide x 68-mm-long Wallstent dilated to 10 mm. TcGSA dynamic imaging studies were performed before and twice after (7 and 14 days) TIPS. To be included in the study, pigs had to have a patent TIPS at 1 week of follow-up. Liver function tests were drawn parallel to the TcGSA studies. Liver biopsies were performed at 2 weeks when the animals were killed. RESULTS: Five of the eight pigs had open shunts at 1 week and were included in the study. There was a significant (P < .0001, Student t test) difference between the mean [R]o of the pre-TIPS studies ([R]o = 1.12+/-0.04 microM) and the mean of the post-TIPS studies at 7 days ([R]o = 0.40+/-0.04 microM) and 14 days ([R]o = 0.51+/-0.06 microM, P < .01). The only blood test that was abnormal after TIPS was ammonia (mean, 129.0+/-42.7). Liver biopsies were normal. CONCLUSIONS: TIPS reduces asialoglycoprotein receptor concentration in normal pigs. PMID- 10693725 TI - Hemodialysis access declotting: a native fistula is not a prosthetic graft. PMID- 10693726 TI - Dextromethorphan: another "ecstasy"? PMID- 10693727 TI - Comorbidity and diagnosing depressive disorders in family practice. PMID- 10693728 TI - Let the buyer (and reader) beware: targeted advertising in medical journals. PMID- 10693729 TI - Postfertilization effects of oral contraceptives and their relationship to informed consent. AB - The primary mechanism of oral contraceptives is to inhibit ovulation, but this mechanism is not always operative. When breakthrough ovulation occurs, then secondary mechanisms operate to prevent clinically recognized pregnancy. These secondary mechanisms may occur either before or after fertilization. Postfertilization effects would be problematic for some patients, who may desire information about this possibility. This article evaluates the available evidence for the postfertilization effects of oral contraceptives and concludes that good evidence exists to support the hypothesis that the effectiveness of oral contraceptives depends to some degree on postfertilization effects. However, there are insufficient data to quantitate the relative contribution of postfertilization effects. Despite the lack of quantitative data, the principles of informed consent suggest that patients who may object to any postfertilization loss should be made aware of this information so that they can give fully informed consent for the use of oral contraceptives. PMID- 10693730 TI - Family responsibilities and domestic activities of US women physicians. AB - BACKGROUND: Women physicians may have a multiplicity of domestic roles (eg, cook, housekeeper, child care provider) that are of inherent interest and that may affect their professional lives, but are largely unstudied. DESIGN, SETTING, PARTICIPANTS, AND MAIN OUTCOME MEASURES: We report data from respondents (N = 4501) to the Women Physicians' Health Study, a cross-sectional, questionnaire based study of a stratified random sample of US women MDs. RESULTS: Women physicians with children aged 0 to 17 years spent a median of 24.4 hours per week on child care. Women physicians typically spent half an hour per day cooking, and another half-hour per day on other housework. Little time was spent on gardening: a median of 0.05 hours (3 minutes) per week. Those performing more domestic tasks are likely to work fewer hours outside the home and to be on call less often. Women physicians who are married or widowed, have more children, have lower personal incomes, and have more highly educated and higher-earning spouses perform more domestic activities. We found no significant adverse relationship between time spent on any domestic activity and career satisfaction or mental or physical health. CONCLUSIONS: Women physicians spend little time on domestic activities that can be done for them by others, including cooking, housework, and especially gardening. Women physicians spend somewhat less time on child care and substantially less time on housework than do other US women. Despite abundant editorializing about role conflicts of women physicians, our measures of career satisfaction and mental health were not adversely affected by time spent on domestic obligations. PMID- 10693731 TI - Self-reported arthritis-related disruptions in sleep and daily life and the use of medical, complementary, and self-care strategies for arthritis: the National Survey of Self-care and Aging. AB - OBJECTIVE: To assess relations between self-reported arthritis-related disruptions in sleep, physical activity, and social functioning and use of medical care, complementary therapies, and self-care for arthritis in older adults. DESIGN: A survey of self-reported arthritis-related disruptions in sleep and daily life as risk factors for use of 15 medical, complementary, and self care modalities for relief of arthritis symptoms. SETTING: General community from 38 urban and 12 rural areas in the contiguous United States. PARTICIPANTS: Nine hundred thirty-seven older persons reporting arthritis; of the 1925 in the 1993 to 1994 follow-up of the National Survey of Self-care and Aging, a population based, stratified, random sample of noninstitutionalized Medicare beneficiaries aged 65 years and older. MAIN OUTCOME MEASURES: Use of 15 medical, self-care, and complementary modalities for relief of arthritis symptoms. RESULTS: Most respondents reported use of at least 1 medical, complementary, or self-care strategy for arthritis. Arthritis was reported to disrupt sleep and leisure in 32.8% and 33.4% of respondents, respectively. Individuals with sleep disruption were more likely than those without sleep disturbance to use medical, complementary, and self-care strategies (adjusted odds ratio [95% confidence interval], 2.31 [1.59-3.37] for seeing a physician; and 2.23 [1.60-3.10] for using physical modalities). Reported disruption in sleep from arthritis was associated with use of more medical, complementary, and self-care strategies than was any other disruption. CONCLUSIONS: Self-reported arthritis-related disruption in sleep is associated with use of a wide range of medical, complementary, and self-care strategies. Physicians, other health care providers, and researchers should not overlook the importance of this common and often-neglected symptom. PMID- 10693732 TI - The role of competing demands in the treatment provided primary care patients with major depression. AB - OBJECTIVE: To examine whether competing demands explain the appearance of inadequate primary care depression treatment observed at a single visit. DESIGN: A cross-sectional patient survey. PARTICIPANTS AND SETTING: Two hundred forty patients with 5 or more symptoms of depression seeing 12 physicians in 6 primary care practices, representing 77.4% of the depressed patients identified through 2 stage screening of more than 11,000 primary care attenders. MAIN OUTCOME MEASURES: In patients with elevated depressive symptoms, discussing depression as a possible diagnosis in untreated patients, and changing depression management in treated patients. RESULTS: Physicians and patients discussed depression in 46 (47.9%) of 96 untreated patients; physicians changed depression treatment recommendations in 87 (60.4%) of 144 treated patients with current symptoms. Chronic physical comorbidity decreased the odds that physicians and untreated patients discussed depression as a possible diagnosis (odds ratio = 0.66, P = .01). New problems decreased the odds that treatment recommendations would be changed in treated patients who remained depressed (odds ratio = 0.39, P = .05). Physicians and untreated patients were more likely to discuss depression as a possible diagnosis if patients reported antidepressant medication was acceptable (odds ratio = 4.57, P = .01) and less likely to discuss depression if patients reported specialty care counseling was acceptable (odds ratio = 0.33, P = .05). CONCLUSIONS: The attention depression gets during a given medical visit is less associated with the severity of the patient's depressive symptoms than with the number or recency of other problems the patient has. If competing demands provide ongoing barriers to depression treatment, interventions will be needed to assure that patients with chronic physical problems receive high-quality mental health care in the primary care setting. PMID- 10693733 TI - Sun protection counseling for children: primary care practice patterns and effect of an intervention on clinicians. AB - OBJECTIVES: To describe current primary care sun protection advice for children and assess the effect on clinicians of an intervention to enhance their sun protection advocacy. SETTING: Primary care practices caring for children in New Hampshire with special attention to clinicians serving 10 towns that were involved in a randomized controlled trial of the multicomponent SunSafe intervention involving schools, recreation areas, and primary care practices. DESIGN/INTERVENTION: A statewide survey of all primary care clinicians serving children addressed their self-reported sun protection advocacy practices. Clinicians in 10 systematically selected rural towns were involved in the subsequent intervention study. The primary care intervention provided assistance to practices in establishing an office system that promoted sun protection advice to children and their parents during office visits. MAIN OUTCOME MEASURES: Sun protection promotion activities of primary care clinicians as determined by their self report, research assistant observation, and parent interviews. RESULTS: Of 261 eligible clinicians responding to the statewide survey, about half provide sun protection counseling "most of the time" or "almost always" during summer well care visits. Pediatricians do so more often than family physicians. Clinicians involved in the intervention increased their use of handouts, waiting room educational materials, and sunscreen samples. Compared with control town parents, parents in intervention towns reported an increase in clinician sun protection advice. CONCLUSIONS: The SunSafe primary care intervention increased sun protection counseling activities of participating clinicians. A single-focus preventive service office system is feasible to include in community interventions to promote sun protection. PMID- 10693734 TI - Orlistat in the long-term treatment of obesity in primary care settings. AB - OBJECTIVE: To evaluate the long-term efficacy and tolerability within primary care settings of orlistat, a gastrointestinal lipase inhibitor, for the treatment of obesity. DESIGN: Randomized, double-blind, placebo-controlled, multicenter study. PARTICIPANTS: A group of 796 obese patients (body mass index, 30-44 kg/m2), treated with placebo 3 times a day (TID), 60 mg of orlistat TID, or 120 mg of orlistat TID, in conjunction with a reduced-energy diet for the first year and a weight-maintenance diet during the second year. SETTING: Seventeen primary care centers in the United States. MAIN OUTCOME MEASURES: Changes in body weight and obesity-related disease risk factors. RESULTS: Patients treated with orlistat lost significantly more weight (7.08 +/- 0.54 and 7.94 +/- 0.57 kg for the 60-mg and 120-mg orlistat groups, respectively) than those treated with placebo (4.14 +/- 0.56 kg) in year 1 (P<.001) and sustained more of this weight loss during year 2 (P<.001). More patients treated with orlistat lost 5% or more of their initial weight in year 1 (48.8% and 50.5% of patients in the 60-mg and 120-mg groups, respectively) compared with placebo (30.7%; P<.001), and approximately 34% of patients in the orlistat groups sustained weight loss of 5% or greater over 2 years compared with 24% in the placebo group (P<.001). Orlistat produced greater improvements than placebo in serum lipid levels and blood pressure and was well tolerated, although treatment resulted in a higher incidence of gastrointestinal events. CONCLUSIONS: This long-term study indicates that orlistat is an effective adjunct to dietary intervention in the treatment of obesity in primary care settings. PMID- 10693735 TI - Unattended home diagnosis and treatment of obstructive sleep apnea without polysomnography. AB - OBJECTIVE: To test the effectiveness of unattended home monitoring along with automatic titrating continuous positive airway pressure (auto-CPAP) as an acceptable method for diagnosing and prescribing proper CPAP pressure for treatment of patients presenting with classic symptoms of obstructive sleep apnea (OSA). DESIGN: Nonrandomized, prospective case study of 63 patients with a presumptive diagnosis of OSA. SETTING: University hospital and veterans affairs medical center ambulatory sleep disorders clinics. PARTICIPANTS: Fifty-eight men and 5 women were recruited for symptoms of excessive daytime sleepiness, heavy snoring, and witnessed apnea. INTERVENTION: Subjects with 10 or more respiratory events per hour were titrated by automatic, unattended home monitoring to an optimal CPAP pressure. MAIN OUTCOME MEASURES: Number of subjects able to be diagnosed by unattended home monitoring, titrated to optimal CPAP pressure, accepted an auto-CPAP machine for home use, and symptoms improved. RESULTS: Fifty four (86%) of 63 patients completed sufficient diagnostic studies, and in 45 (83%) of these, a diagnosis of OSA was established. Nine subjects were unable to adjust to the nasal mask for an adequate diagnostic recording, and 9 had fewer than 10 respiratory events per hour. Ten subjects with OSA could not complete a titration study. Thirty-five of the subjects diagnosed with OSA accepted the auto CPAP machine into their home, while 30 used it for therapy longer than 3 weeks. The estimated cost of performing in-home studies was less than one fourth of the estimated cost for in-laboratory polysomnographic examinations had they been performed. CONCLUSION: Unattended monitoring plus auto-CPAP allows cost-effective diagnosis and CPAP titration of most patients with OSA with straightforward symptoms. PMID- 10693736 TI - New treatments and therapeutic strategies for acne. AB - Successful management of acne requires careful patient evaluation followed by consideration of several patient and medication factors when selecting a particular therapeutic regimen. Within the last few years, several new agents for the treatment of acne have become available that afford greater flexibility in the treatment of this prevalent dermatologic disorder. These include adapalene, tazarotene, 2 new topical tretinoin formulations, azelaic acid, a new sodium sulfacetamide formulation, and an oral contraceptive recently approved by the Food and Drug Administration for the treatment of acne. After a brief overview of the pathophysiology of acne and existing therapies, this review evaluates the new antiacne agents and how they can be integrated into a successful treatment strategy that takes into account acne severity and predominant lesion type as well as age, skin type, lifestyle, motivation, and the presence of coexisting conditions. PMID- 10693737 TI - Hypertriglyceridemia and coronary heart disease. PMID- 10693738 TI - Improving social adjustment in children with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: To determine if sending motivational letters would improve peer relations in children with social maladjustment and attention deficit/hyperactivity disorder (ADHD). DESIGN: From a consecutive sample, a case series was followed up for 2 years. SETTING: Primary care, private physician, office-based practice. PATIENTS: Ninety-five children diagnosed as having ADHD by Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria made up the consecutive sample. Twenty-one children qualified with a comorbid social maladjustment disorder with baseline t scores of 1.5 SDs or greater above the mean on the asocial domain of the Conners' Teacher Rating Scale. Seventeen children completed the study. There were no significant differences between these patients and those who did not complete the study (P = .55 for baseline score differences and P = .85 for age). INTERVENTIONS: In addition to conventional therapy for ADHD when children achieved a goal, such as an improved report card or better conduct, a personal letter about their success was mailed to them. Letters averaged 5 per student per year. MAIN OUTCOME MEASURES: The asocial domain of the Conners' Teacher Rating Scale was repeated during the next 2 school years for comparison. Statistical analysis was by a repeated-measures analysis of variance and Helmert contrasts. RESULTS: Of 17 students who completed the study, 16 improved on the Conners' Teacher Rating Scale asocial domain, and the results were statistically significant (P<.001). CONCLUSIONS: Difficulties with peer relations are commonly seen in children with ADHD. Sending motivational letters correlated with improved social adjustment in these children. The data suggest that busy practitioners might consider incorporating this successful, and time efficient, intervention into their ADHD treatment regimens. PMID- 10693739 TI - Spontaneous splenic infarction secondary to diabetes-induced microvascular disease. AB - Splenic infarction is a clinical entity seldom encountered. The most frequent causes of splenic infarction include thromboembolic phenomena, hematologic malignant neoplasms, and vasculitides. We describe a patient who sustained splenic infarction secondary to diabetes-induced, small-vessel atherosclerotic disease. PMID- 10693740 TI - A product of the culture. PMID- 10693742 TI - Characterization of Trypanozoon isolates using a repeated coding sequence and microsatellite markers. AB - Genetic variation of microsatellite loci is a widely used method for linkage analysis, individual identification or inter-population studies. Here we analyse a repeated DNA coding sequence and eleven new microsatellites identified within the Trypanosoma (Trypanozoon) brucei genome. Ninety-seven isolates belonging to the five species and subspecies Trypanosoma evansi, T. equiperdum, T. brucei brucei, T. b. rhodesiense and T. b. gambiense were compared regarding the genetic patterns of these markers. The results reveal a great heterogeneity of the genotypes related to the repeated coding sequence and five microsatellites, some of which show a high degree of polymorphism. This allows us to define group specific genotypes or alleles; in particular, we show that one specific pattern clearly segregates the human pathogen T. b. gambiense group I. PMID- 10693741 TI - Molecular characterization of P-29, a metacestode-specific component of Echinococcus granulosus which is immunologically related to, but distinct from, antigen 5. AB - In this work the characterization of P-29, a novel 29 kDa antigen from Echinococcus granulosus is reported. E. granulosus was identified while looking for parasite antigens distinct from those present in hydatid cyst fluid. A monoclonal antibody (mAb 47H.PS) prepared against protoscolex components revealed that P-29 is localized to the tegument and rostellum of protoscoleces, and to the germinal layer of the cyst, but it is absent in hydatid cyst fluid or adult worm extracts. Several internal fragments of P-29 showed sequence identity to the amino acid sequence encoded by Eg6, a partial gene sequence reported to code for an epitope of antigen 5 (Ag5), one of the major diagnostic antigens of the parasite. We confirmed that Eg6 encodes a sub-fragment of P-29 by mapping the epitope of mAb 47H.PS, and isolating the full length P-29 cDNA. Since Eg6 had been, postulated to encode a fragment of Ag5, we specifically studied the relationship of P-29 and Ag5 by: (i) examining the cross-reactivity displayed by different mAbs; (ii) comparison of their peptide finger prints; and (iii) a comparative study of their diagnostic value. Our results prove unequivocally that P-29 and Ag5 are immunologically related, but different proteins, raising several questions on the current knowledge of Ag5. PMID- 10693743 TI - Tetrameric and dimeric malate dehydrogenase isoenzymes in Trypanosoma cruzi epimastigotes. AB - Two malate dehydrogenase isoforms, named MDH1 and MDH2, have been purified to homogeneity from Trypanosoma cruzi epimastigotes. Both enzymes consist of subunits with a molecular mass close to 33 kDa; native molecular mass determination by gel filtration, however, indicated that MDH1 is a dimer, whereas MDH2 is a tetramer. Both isoforms did not cross-react immunologically. The N termini of both MDH isoforms and several tryptic peptides of MDH1 (amounting to about one third of the complete molecule) have been sequenced by automated Edman degradation. The tryptic digests of both enzymes have also been analysed by mass spectrometry (MALDI-TOF MS). The apparent Km values in both directions of the reaction have been determined, as well as the possible inhibition by excess of the substrate oxaloacetate. The sequence data, together with the pI values and the presence or absence of oxaloacetate inhibition indicate that the dimeric MDH1 is the mitochondrial isoenzyme, whereas the tetrameric MDH2 is the glycosomal isoenzyme. No evidence was found for the presence of a cytosolic isoform. PMID- 10693744 TI - Purification and characterization of Plasmodium falciparum succinate dehydrogenase. AB - Succinate dehydrogenase (SDH), a Krebs cycle enzyme and complex II of the mitochondrial electron transport system was purified to near homogeneity from the human malarial parasite Plasmodium falciparum cultivated in vitro by FPLC on Mono Q, Mono S and Superose 6 gel filtration columns. The malarial SDH activity was found to be extremely labile. Based on Superose 6 FPLC, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing-PAGE analyses, it was demonstrated that the malarial enzyme had an apparent native molecular mass of 90 +/- 8 kDa and contained two major subunits with molecular masses of 55 +/- 6 and 35 +/- 4 kDa (n = 8). The enzymatic reaction required both succinate and coenzyme Q (CoQ) for its maximal catalysis with Km values of 3 and 0.2 microM, and k(cat) values of 0.11 and 0.06 min(-1), respectively. Catalytic efficiency of the malarial SDH for both substrates were found to be relatively low (approximately 600-5000 M(-1) s(-1)). Fumarate, malonate and oxaloacetate were found to inhibit the malarial enzyme with Ki values of 81, 13 and 12 microM, respectively. The malarial enzyme activity was also inhibited by substrate analog of CoQ, 5-hydroxy-2-methyl-1,4-naphthoquinone, with a 50% inhibitory concentration of 5 microM. The quinone had antimalarial activity against the in vitro growth of P. falciparum with a 50% inhibitory concentration of 0.27 microM and was found to completely inhibit oxygen uptake of the parasite at a concentration of 0.88 microM. A known inhibitor of mammalian mitochondrial SDH, 2 thenoyltrifluoroacetone. had no inhibitory effect on both the malarial SDH activity and the oxygen uptake of the parasite at a concentration of 50 microM. Many properties observed in the malarial SDH were found to be different from the host mammalian enzyme. PMID- 10693745 TI - Secretion of a novel class of iFABPs in nematodes: coordinate use of the Ascaris/Caenorhabditis model systems. AB - A novel fatty acid binding protein, As-p18, is secreted into both the perivitelline and perienteric fluids of the parasitic nematode, Ascaris suum, and at least eight potential homologues of As-p18 have been identified in the Caenorhabditis elegans genome. The products of the three most closely related homologues are fatty acid binding proteins (LBP-1, LBP-2 and LBP-3) which contain putative secretory signals. Phylogenetic analysis revealed that these secreted fatty acid binding proteins comprise a distinct gene class within the fatty acid binding protein family and are possibly unique to nematodes. To examine the potential sites of As-p18 secretion, the expression of the putative promoters of the C. elegans homologues was examined with GFP reporter constructs. The developmental expression of lbp-1 was identical to that of As-p18 and consistent with the secretion of LBP-1 from the hypodermis to the perivitelline fluid. The expression patterns of lbp-2 and lbp-3 were consistent with the secretion of LBP 2 and LBP-3 from muscle into the perienteric fluid later in development. These studies demonstrate that at least some perivitelline fluid proteins appear to be secreted from the hypodermis prior to the formation of the cuticle and, perhaps more importantly, that this coordinate C. elegans/A. suum approach may be potentially useful for examining a number of key physiological processes in parasitic nematodes. PMID- 10693746 TI - Intracellular pH in mammalian stages of Trypanosoma cruzi is K+-dependent and regulated by H+-ATPases. AB - Regulation of intracellular pH (pHi) was investigated in Trypanosoma cruzi amastigotes and trypomastigotes using 2',7'-bis-(carboxyethyl)-5(and-6) carboxyfluorescein (BCECF). pHi was determined to be 7.33 +/- 0.08 and 7.35 +/- 0.07 in amastigotes and trypomastigotes, respectively, and there were no significant differences in the regulation of pH, between the two stages. Steady state pHi, recovery of pHi from acidification, and H+-efflux were all decreased markedly by the H+-ATPase inhibitors N,N'-dicyclohexylcarbodi-imide (DCCD), diethylstilbestrol (DES) and N-ethylmaleimide (NEM) supporting a significant role for a plasma membrane H+-ATPase in the regulation of pHi. pHi was maintained at neutrality over a range of external pH (pHe) from 5-8 in parasites suspended in a buffer containing Na+ and K+ (standard buffer) but was acidified at low pHe in the absence of these cations (choline buffer). The pHi of trypomastigotes decreased significantly when they transformed into amastigotes. The rate of recovery of pHi by acidified parasites was similar in Na+-free buffer and standard buffer but was slower in the absence of K+ (K+-free or choline buffer) and parasites suspended in choline buffer were acidic by 0.25 pH units as compared with controls. Ba2+ and Cs+ decreased the pHi of parasites suspended in standard but not choline buffer suggesting the presence of an inward directed K+ channel. The pHi of amastigotes and trypomastigotes suspended in Cl(-)-free buffer was decreased by 0.13 and 0.2 pH units, respectively, supporting the presence of a chloride conductive channel. No evidence of pH regulation via a Na+/H+ or Cl-/HCO3- exchanger was found. These results are consistent with the presence of a plasma membrane H+-ATPase that regulates pHi and is supported by K+ and Cl- channels. PMID- 10693747 TI - Molecular analysis of a Type I fatty acid synthase in Cryptosporidium parvum. AB - We report here the molecular analysis of a Type I fatty acid synthase in the apicomplexan Cryptosporidium parvum (CpFAS1). The CpFAS1 gene encodes a multifunctional polypeptide of 8243 amino acids that contains 21 enzymatic domains. This CpFAS1 structure is distinct from that of mammalian Type I FAS, which contains only seven enzymatic domains. The CpFAS1 domains are organized into: (i) a starter unit consisting of a fatty acid ligase and an acyl carrier protein; (ii) three modules, each containing a complete set of six enzymes (acyl transferase, ketoacyl synthase, ketoacyl reductase, dehydrase, enoyl reductase, and acyl carrier protein) for the elongation of fatty acid C2-units; and (iii) a terminating domain whose function is as yet unknown. The CpFAS1 gene is expressed throughout the life cycle of C. parvum, since its transcripts and protein were detected by RT-PCR and immunofluorescent localization, respectively. This cytosolic Type I CpFAS1 differs from the organellar Type II FAS enzymes identified from Toxoplasma gondii and Plasmodium falciparum which are targetted to the apicoplast, and are sensitive to inhibition by thiolactomycin. That the discovery of CpFAS1 may provide a new biosynthetic pathway for drug development against cryptosporidiosis, is indicated by the efficacy of the FAS inhibitor cerulenin on the growth of C. parvum in vitro. PMID- 10693748 TI - Calcium mobilization by arachidonic acid in trypanosomatids. AB - A recent report (Eintracht J, Maathai R, Mellors A, Ruben L. Calcium entry in Trypanosoma brucei is regulated by phospholipase A, and arachidonic acid, Biochem J 1998:336:659-66) provided evidence that calcium entry in Trypanosoma brucei bloodstream trypomastigotes is regulated via a signaling pathway involving phospholipase A2-mediated generation of arachidonic acid and stimulation of a plasma membrane-located calcium channel. Here we show that Ca2+ influx in T. brucei procyclic trypomastigotes, Leishmania donovani promastigotes and T. cruzi amastigotes was also stimulated in a dose-dependent manner (50-400 nM) by the amphiphilic peptide melittin. This effect was blocked by the phospholipase A, inhibitor 3-(4-octadecyl)-benzoylacrylic acid. The unsaturated fatty acid arachidonic acid, in the range of 10-75 microM, induced Ca2+ entry by a mechanism sensitive to LaCl3. However, both melittin and arachidonic acid induced an increase in [Ca2+]i in T. brucei procyclic trypomastigotes incubated in Ca2+-free medium implying Ca2+ mobilization from intracellular stores. This hypothesis was supported by experiments showing that arachidonic acid promoted Ca2+ release from the acidocalcisomes of these cells. The results showing changes in mitochondrial membrane potential, release of acridine orange and Ca2+ from the acidocalcisomes and Ca2+ transport across the plasma membrane suggest that in addition to the possible stimulation of a Ca2+ channel-mediated process, arachidonic acid, in the range of concentrations used here, have other nonspecific effects on the trypanosomatids membranes. PMID- 10693749 TI - The secretory Onchocerca volvulus protein OvS1/Ov20 exhibits the capacity to compete with serum albumin for the host's long-chain fatty acids. AB - The mechanism by which filarial parasites derive fatty acids bound to the host's carrier protein is poorly understood. The capacity of a secretory protein of Onchocerca volvulus (OvS1/Ov20) to compete with serum albumin for arachidonic and other fatty acids was investigated in this study. Binding affinities of the two proteins for the long-chain fatty acids were determined using displacement assays. The fluorescent probes used included 11-((5-dimethylaminonaphthalene-1 sulfonyl)amino) undecanoic acid (DAUDA) and cis-parinaric acid. OvS1 protein bound arachidonic acid with an affinity five-fold greater than the affinity exhibited by serum albumin. Oleic acid was bound by the parasite protein with an affinity two-fold greater than the affinity shown by serum albumin. Furthermore, the affinities exhibited by OvS1 protein in binding arachidonic and linoleic acid were about two times higher than the affinity for oleic acid. The results suggest that the OvS1 protein has the capacity to compete with the main host's fatty acid carrier protein for the long-chain fatty acids, in particular arachidonic acid, the precursor for eicosanoids. PMID- 10693750 TI - Characterisation of the loci encoding the glutamic acid and alanine rich protein of Trypanosoma congolense. AB - We have characterised the organisation of genes encoding the glutamate and alanine rich protein (GARP) surface coat of the procyclic and epimastigote stages of Trypanosoma congolense in the tsetse fly. The GARP genes are arranged at two, possibly physically linked, loci, one of which exhibits allelic variation. One locus contains a single GARP gene, whilst both alleles of the other have a large tandem array of polycistronically transcribed GARP genes. Sequence analysis has revealed that there are very few coding differences between different GARP genes. A sequence related to the Trypanosoma brucei expression site associated gene 4 (encoding a transmembrane protein with a cytoplasmic adenylate cyclase domain) has been identified within a region at the downstream flank of one locus. There is no evidence that, within the single trypanosome, GARP genes are as diverse as the procyclin genes that encode a corresponding coat in T. brucei. PMID- 10693751 TI - Fluorescence microscopy and fluorescence in situ hybridization of Entamoeba histolytica nuclei to analyse mitosis and the localization of repetitive DNA. PMID- 10693752 TI - A novel member of the NK-lysin protein family is developmentally regulated and secreted by Fasciola hepatica. PMID- 10693753 TI - Two major 'higher molecular mass proteinases' of Entamoeba histolytica are identified as cysteine proteinases 1 and 2. PMID- 10693754 TI - A simple RNA analysis method shows var and rif multigene family expression patterns in Plasmodium falciparum. PMID- 10693755 TI - The multiple roles of PTEN in tumor suppression. PMID- 10693756 TI - Regulated intramembrane proteolysis: a control mechanism conserved from bacteria to humans. PMID- 10693757 TI - Analysis of partner of inscuteable, a novel player of Drosophila asymmetric divisions, reveals two distinct steps in inscuteable apical localization. AB - Asymmetric localization is a prerequisite for inscuteable (insc) to function in coordinating and mediating asymmetric cell divisions in Drosophila. We show here that Partner of Inscuteable (Pins), a new component of asymmetric divisions, is required for Inscuteable to asymmetrically localize. In the absence of pins, Inscuteable becomes cytoplasmic and asymmetric divisions of neuroblasts and mitotic domain 9 cells show defects reminiscent of insc mutants. Pins colocalizes with Insc and interacts with the region necessary and sufficient for directing its asymmetric localization. Analyses of pins function in neuroblasts reveal two distinct steps for Insc apical cortical localization: A pins-independent, bazooka dependent initiation step during delamination (interphase) and a later maintenance step during which Baz, Pins, and Insc localization are interdependent. PMID- 10693758 TI - Opposing transcriptional outputs of Hedgehog signaling and engrailed control compartmental cell sorting at the Drosophila A/P boundary. AB - The wing imaginal disc is subdivided into two nonintermingling sets of cells, the anterior (A) and posterior (P) compartments. Anterior cells require reception of the Hedgehog (Hh) signal to segregate from P cells. We provide evidence that Hh signaling controls A/P cell segregation not by directly modifying structural components but by a Cubitus interruptus (Ci)-mediated transcriptional response. A shift in the balance between repressor and activator forms of Ci toward the activator form is necessary and sufficient to define "A-type" cell sorting behavior. Moreover, we show that Engrailed (En), in the absence of Ci, is sufficient to specify "P-type" sorting. We propose that the opposing transcriptional activities of Ci and En control cell segregation at the A/P boundary by regulating a single cell adhesion molecule. PMID- 10693759 TI - Hedgehog-regulated processing of Gli3 produces an anterior/posterior repressor gradient in the developing vertebrate limb. AB - Ci/Gli zinc finger proteins mediate the transcriptional effects of Hedgehog protein signals. In Drosophila, Ci action as transcriptional repressor or activator is contingent upon Hedgehog-regulated, PKA-dependent proteolytic processing. We demonstrate that PKA-dependent processing of vertebrate Gli3 in developing limb similarly generates a potent repressor in a manner antagonized by apparent long-range signaling from posteriorly localized Sonic hedgehog protein. The resulting anterior/posterior Gli3 repressor gradient can be perturbed by mutations of Gli3 in human genetic syndromes or by misregulation of Gli3 processing in the chicken mutant talpid2, producing a range of limb patterning malformations. The high relative abundance and potency of Gli3 repressor suggest specialization of Gli3 and its products for negative Hedgehog pathway regulation. PMID- 10693760 TI - Ras1 promotes cellular growth in the Drosophila wing. AB - The Ras GTPase links extracellular mitogens to intracellular mechanisms that control cell proliferation. To understand how Ras regulates proliferation in vivo, we activated or inactivated Ras in cell clones in the developing Drosophila wing. Cells lacking Ras were smaller, had reduced growth rates, accumulated in G1, and underwent apoptosis due to cell competition. Conversely, activation of Ras increased cell size and growth rates and promoted G1/S transitions. Ras upregulated the growth driver dMyc, and both Ras and dMyc increased levels of cyclin E posttranscriptionally. We propose that Ras primarily promotes growth and that growth is coupled to G1/S progression via cyclin E. Interestingly, upregulation of growth by Ras did not deregulate G2/M progression or a developmentally regulated cell cycle exit. PMID- 10693761 TI - Crystal structure of the VHS and FYVE tandem domains of Hrs, a protein involved in membrane trafficking and signal transduction. AB - We have determined the 2 A X-ray structure of the 219-residue N-terminal VHS and FYVE tandem domain unit of Drosophila Hrs. The unit assumes a pyramidal structure in which the much larger VHS domain (residues 1-153) forms a rectangular base and the FYVE domain occupies the apical end. The VHS domain is comprised of an unusual "superhelix" of eight alpha helices, and the FYVE domain is mainly built of loops, two double-stranded antiparallel sheets, and a helix stabilized by two tetrahedrally coordinated zinc atoms. The two-domain structure forms an exact 2 fold-related homodimer through antiparallel association of mainly FYVE domains. Dimerization creates two identical pockets designed for binding ligands with multiple negative charges such as citrate or phosphatidylinositol 3-phosphate. PMID- 10693762 TI - A role for CaMKII in T cell memory. AB - In order to study the role of calcium/calmodulin kinase II (CaMKII) in T cells, we generated transgenic mice expressing CaMKIIgammaB* (T287D), a partially calcium-independent mutant of CaMKIIgammaB. In these mice, the size of the thymus was increased 1.5- to 2-fold, at least in part due to an increase in the lifespan of double-positive (DP) thymocytes. More importantly, there was an increase in the number of T cells in the secondary lymphoid organs that had acquired an antigen-dependent memory phenotype. These T cells were bonafide memory cells as assessed by a variety of criteria. In addition, T cells from wild-type mice acquired calcium-independent CaMKII activity after several rounds of antigen stimulated division. We propose that CaMKII controls a distinct process of activation-induced cellular differentiation. PMID- 10693763 TI - Regulation of auxin response by the protein kinase PINOID. AB - Arabidopsis plants carrying mutations in the PINOID (PID) gene have a pleiotropic shoot phenotype that mimics that of plants grown on auxin transport inhibitors or of plants mutant for the auxin efflux carrier PINFORMED (PIN), with defects in the formation of cotyledons, flowers, and floral organs. We have cloned PID and find that it is transiently expressed in the embryo and in initiating floral anlagen, demonstrating a specific role for PID in promoting primordium development. Constitutive expression of PID causes a phenotype in both shoots and roots that is similar to that of auxin-insensitive plants, implying that PID, which encodes a serine-threonine protein kinase, negatively regulates auxin signaling. PMID- 10693764 TI - The Saccharomyces Pif1p DNA helicase and the highly related Rrm3p have opposite effects on replication fork progression in ribosomal DNA. AB - Replication of Saccharomyces ribosomal DNA (rDNA) proceeds bidirectionally from origins in a subset of the approximately 150 tandem repeats, but the leftward moving fork stops when it encounters the replication fork barrier (RFB). The Pif1p helicase and the highly related Rrm3p were rDNA associated in vivo. Both proteins affected rDNA replication but had opposing effects on fork progression. Pif1p helped maintain the RFB. Rrm3p appears to be the replicative helicase for rDNA as it acted catalytically to promote fork progression throughout the rDNA. Loss of Rrm3p increased rDNA breakage and accumulation of rDNA circles, whereas breakage and circles were less common in pif1 cells. These data support a model in which replication fork pausing causes breakage and recombination in the rDNA. PMID- 10693765 TI - Science and more for TB. PMID- 10693766 TI - Fossil smuggling unopposed. PMID- 10693767 TI - Feathers fly over Chinese fossil bird's legality and authenticity. PMID- 10693768 TI - The biggest, wildest fossil market in the west. PMID- 10693769 TI - Protein structure groups seek to draft common ground rules. PMID- 10693770 TI - South African government rejects AZT advice. PMID- 10693771 TI - Consortium aims to kick-start TB research. PMID- 10693772 TI - Europe's X-ray observatory defies the jinx... PMID- 10693773 TI - Oxford professor faces business link inquiry. PMID- 10693774 TI - Reliance on the citation index undermines the study of biodiversity. PMID- 10693775 TI - Proteomics could be key in battle against malaria. PMID- 10693776 TI - X chromosomes forget where they came from. PMID- 10693777 TI - Don't let politics put Diamond at risk PMID- 10693778 TI - One-stop shop for microarray data. PMID- 10693779 TI - Where do babies come from? PMID- 10693780 TI - Total internal reflection. PMID- 10693781 TI - Taiwan's gift to the world. PMID- 10693782 TI - Display technology. Sidestepping the selection rules PMID- 10693784 TI - Climate change. The hole record PMID- 10693783 TI - Mining the genome for iron. PMID- 10693785 TI - A provirus put to work. PMID- 10693786 TI - Superconductivity. From obscurity to impurity PMID- 10693787 TI - Ecology. The complexity of co-dependency. PMID- 10693788 TI - Mathematics. A gathering of groups PMID- 10693790 TI - Dennis Sciama (1926-99) PMID- 10693789 TI - Developmental biology. Raising the roof. PMID- 10693791 TI - Conversion of diploidy to haploidy. PMID- 10693792 TI - Reactivation of Borrelia infection in birds. PMID- 10693793 TI - Do mussels take wooden steps to deep-sea vents? PMID- 10693794 TI - Supernova explosions in the Universe. AB - During the lifetime of our Milky Way galaxy, there have been something like 100 million supernova explosions, which have enriched the Galaxy with the oxygen we breathe, the iron in our cars, the calcium in our bones and the silicon in the rocks beneath our feet. These exploding stars also influence the birth of new stars and are the source of the energetic cosmic rays that irradiate us on the Earth. The prodigious amount of energy (approximately 10(51), or approximately 2.5 x 10(28) megatonnes of TNT equivalent) and momentum associated with each supernova may even have helped to shape galaxies as they formed in the early Universe. Supernovae are now being used to measure the geometry of the Universe, and have recently been implicated in the decades-old mystery of the origin of the gamma-ray bursts. Together with major conceptual advances in our theoretical understanding of supernovae, these developments have made supernovae the centre of attention in astrophysics. PMID- 10693795 TI - Genetic ablation reveals that the roof plate is essential for dorsal interneuron specification. AB - During neural development in vertebrates, a spatially ordered array of neurons is generated in response to inductive signals derived from localized organizing centres. One organizing centre that has been proposed to have a role in the control of neural patterning is the roof plate. To define the contribution of signals derived from the roof plate to the specification of neuronal cell types in the dorsal neural tube, we devised a genetic strategy to ablate the roof plate selectively in mouse embryos. Embryos without a roof plate lack all the interneuron subtypes that are normally generated in the dorsal third of the neural tube. Using a genetically based lineage analysis and in vitro assays, we show that the loss of these neurons results from the elimination of non autonomous signals provided by the roof plate. These results reveal that the roof plate is essential for specifying multiple classes of neurons in the mammalian central nervous system. PMID- 10693796 TI - Origin of the Moon's orbital inclination from resonant disk interactions AB - The Moon is generally believed to have formed from the debris disk created by a large body colliding with the early Earth. Recent models of this process predict that the orbit of the newly formed Moon should be in, or very near, the Earth's equatorial plane. This prediction, however, is at odds with the known history of the lunar orbit: the orbit is currently expanding, but can be traced back in time to reveal that, when the Moon formed, its orbital inclination relative to the Earth's equator was I approximately = 10 degrees. The cause of this initial inclination has been a mystery for over 30 years, as most dynamical processes (such as those that act to flatten Saturn's rings) will tend to decrease orbital inclinations. Here we show that the Moon's substantial orbital inclination is probably a natural result of its formation from an impact-generated disk. The mechanism involves a gravitational resonance between the Moon and accretion-disk material, which can increase orbital inclinations up to approximately 15 degrees. PMID- 10693797 TI - Preparing pure photon number states of the radiation field AB - The quantum mechanical description of a radiation field is based on states that are characterized by the number of photons in a particular mode; the most basic quantum states are those with fixed photon number, usually referred to as number (or Fock) states. Although Fock states of vibrational motion can be observed readily in ion traps, number states of the radiation field are very fragile and difficult to produce and maintain. Single photons in multi-mode fields have been generated using the technique of photon pairs. But in order to generate these states in a cavity, the mode in question must have minimal losses; moreover, additional sources of photon number fluctuations, such as the thermal field, must be eliminated. Here we observe the build-up of number states in a high-Q cavity, by investigating the interaction dynamics of a probe atom with the field. We employ a dynamical method of number state preparation that involves state reduction of highly excited atoms in a cavity, with a photon lifetime as high as 0.2 seconds. (This set-up is usually known as the one-atom maser or 'micromaser'.) Pure states containing up to two photons are measured unambiguously. PMID- 10693798 TI - Imaging the effects of individual zinc impurity atoms on superconductivity in Bi2Sr2CaCu2O8+delta AB - Although the crystal structures of the copper oxide high-temperature superconductors are complex and diverse, they all contain some crystal planes consisting of only copper and oxygen atoms in a square lattice: superconductivity is believed to originate from strongly interacting electrons in these CuO2 planes. Substituting a single impurity atom for a copper atom strongly perturbs the surrounding electronic environment and can therefore be used to probe high temperature superconductivity at the atomic scale. This has provided the motivation for several experimental and theoretical studies. Scanning tunnelling microscopy (STM) is an ideal technique for the study of such effects at the atomic scale, as it has been used very successfully to probe individual impurity atoms in several other systems. Here we use STM to investigate the effects of individual zinc impurity atoms in the high-temperature superconductor Bi2Sr2CaCu2O8+delta. We find intense quasiparticle scattering resonances at the Zn sites, coincident with strong suppression of superconductivity within approximately 15 A of the scattering sites. Imaging of the spatial dependence of the quasiparticle density of states in the vicinity of the impurity atoms reveals the long-sought four-fold symmetric quasiparticle 'cloud' aligned with the nodes of the d-wave superconducting gap which is believed to characterize superconductivity in these materials. PMID- 10693799 TI - High-efficiency fluorescent organic light-emitting devices using a phosphorescent sensitizer AB - To obtain the maximum luminous efficiency from an organic material, it is necessary to harness both the spin-symmetric and anti-symmetric molecular excitations (bound electron-hole pairs, or excitons) that result from electrical pumping. This is possible if the material is phosphorescent, and high efficiencies have been observed in phosphorescent organic light-emitting devices. However, phosphorescence in organic molecules is rare at room temperature. The alternative radiative process of fluorescence is more common, but it is approximately 75% less efficient, due to the requirement of spin-symmetry conservation. Here, we demonstrate that this deficiency can be overcome by using a phosphorescent sensitizer to excite a fluorescent dye. The mechanism for energetic coupling between phosphorescent and fluorescent molecular species is a long-range, non-radiative energy transfer: the internal efficiency of fluorescence can be as high as 100%. As an example, we use this approach to nearly quadruple the efficiency of a fluorescent red organic light-emitting device. PMID- 10693800 TI - Explanation for fracture spacing in layered materials AB - The spacing of opening-mode fractures in layered materials--such as certain sedimentary rocks and laminated engineering materials--is often proportional to the thickness of the fractured layer. Experimental studies of this phenomenon show that the spacing initially decreases as extensional strain increases in the direction perpendicular to the fractures. But at a certain ratio of spacing to layer thickness, no new fractures form and the additional strain is accommodated by further opening of existing fractures: the spacing then simply scales with layer thickness, which is called fracture saturation. This is in marked contrast to existing theories of fracture, such as the stress-transfer theory, which predict that spacing should decrease with increasing strain ad infinitum. Recently, two of us (T.B. and D.D.P.) have used a combination of numerical simulations and laboratory experiments to show that, with increasing applied stress, the normal stress acting between such fractures undergoes a transition from tensile to compressive, suggesting a cause for fracture saturation. Here we investigate the full stress distribution between such fractures, from which we derive an intuitive physical model of the process of fracture saturation. Such a model should find wide applicability, from geosciences to engineering. PMID- 10693801 TI - Temperature trends over the past five centuries reconstructed from borehole temperatures AB - For an accurate assessment of the relative roles of natural variability and anthropogenic influence in the Earth's climate, reconstructions of past temperatures from the pre-industrial as well as the industrial period are essential. But instrumental records are typically available for no more than the past 150 years. Therefore reconstructions of pre-industrial climate rely principally on traditional climate proxy records, each with particular strengths and limitations in representing climatic variability. Subsurface temperatures comprise an independent archive of past surface temperature changes that is complementary to both the instrumental record and the climate proxies. Here we use present-day temperatures in 616 boreholes from all continents except Antarctica to reconstruct century-long trends in temperatures over the past 500 years at global, hemispheric and continental scales. The results confirm the unusual warming of the twentieth century revealed by the instrumental record, but suggest that the cumulative change over the past five centuries amounts to about 1 K, exceeding recent estimates from conventional climate proxies. The strength of temperature reconstructions from boreholes lies in the detection of long-term trends, complementary to conventional climate proxies, but to obtain a complete picture of past warming, the differences between the approaches need to be investigated in detail. PMID- 10693802 TI - Effect of stream channel size on the delivery of nitrogen to the Gulf of Mexico AB - An increase in the flux of nitrogen from the Mississippi river during the latter half of the twentieth century has caused eutrophication and chronic seasonal hypoxia in the shallow waters of the Louisiana shelf in the northern Gulf of Mexico. This has led to reductions in species diversity, mortality of benthic communities and stress in fishery resources. There is evidence for a predominantly anthropogenic origin of the increased nitrogen flux, but the location of the most significant sources in the Mississippi basin responsible for the delivery of nitrogen to the Gulf of Mexico have not been clearly identified, because the parameters influencing nitrogen-loss rates in rivers are not well known. Here we present an analysis of data from 374 US monitor ing stations, including 123 along the six largest tributaries to the Mississippi, that shows a rapid decline in the average first-order rate of nitrogen loss with channel size- from 0.45 day (-1) in small streams to 0.005 day (-1) in the Mississippi river. Using stream depth as an explanatory variable, our estimates of nitrogen-loss rates agreed with values from earlier studies. We conclude that the proximity of sources to large streams and rivers is an important determinant of nitrogen delivery to the estuary in the Mississippi basin, and possibly also in other large river basins. PMID- 10693803 TI - Producer-decomposer co-dependency influences biodiversity effects. AB - Producers, such as plants and algae, acquire nutrients from inorganic sources that are supplied primarily by decomposers whereas decomposers, mostly fungi and bacteria, acquire carbon from organic sources that are supplied primarily by producers. This producer-decomposer co-dependency is important in governing ecosystem processes, which implies that the impacts of declining biodiversity on ecosystem functioning should be strongly influenced by this process. Here we show, by simultaneously manipulating producer (green algal) and decomposer (heterotrophic bacterial) diversity in freshwater microcosms, that algal biomass production varies considerably among microcosms (0.0-0.67 mg ml(-1)), but that neither algal nor bacterial diversity by itself can explain this variation. Instead, production is a joint function of both algal and bacterial diversity. Furthermore, the range in algal production in microscosms in which bacterial diversity was manipulated was nearly double (1.82 times) that of microcosms in which bacterial diversity was not manipulated. Measures of organic carbon use by bacteria in these microcosms indicate that carbon usage is the mechanism responsible for these results. Because both producer and microbial diversity respond to disturbance and habitat modification, the main causes of biodiversity loss, these results suggest that ecosystem response to changing biodiversity is likely to be more complex than other studies have shown. PMID- 10693804 TI - The mouse Dreher gene Lmx1a controls formation of the roof plate in the vertebrate CNS. AB - In the vertebrate central nervous system (CNS), a cascade of signals that originates in the ectoderm adjacent to the neural tube is propagated by the roof plate to dorsalize the neural tube. Here we report that the phenotype of the spontaneous neurological mutant mouse dreher (dr) results from a failure of the roof plate to develop. Dorsalization of the neural tube is consequently affected: dorsal interneurons in the spinal cord and granule neurons in the cerebellar cortex are lost, and the dorsal vertebral neural arches fail to form. Positional cloning of dreher indicates that the LIM homeodomain protein, Lmx1a, is affected in three different alleles of dreher. Lmx1a is expressed in the roof plate along the neuraxis during development of the CNS. Thus, Lmx1a is required for development of the roof plate and, in turn, for specification of dorsal cell fates in the CNS and developing vertebrae. PMID- 10693805 TI - Forebrain peptides modulate sexually polymorphic vocal circuitry. AB - The peptide arginine-vasopressin (mammals) and its evolutionary precursor arginine-vasotocin (non-mammals) modulate reproductive physiology and numerous related social behaviours, as do oxytocin (mammals) and its homologues mesotocin and isotocin (fish). The distributions in the brain and/or the behavioural functions of these peptides often differ between the sexes, and between species with divergent social structures. Here we present neurophysiological evidence that males with vocal characteristics typical of females share a pattern of neuropeptide function with females rather than conspecific males. The plainfin midshipman fish (Porichthys notatus) has two male morphs with different reproductive tactics and vocalizations (a key species-typical behaviour which varies in its physical attributes and contextual usage, depending on the morph's social strategy). Forebrain-evoked, rhythmic vocal-motor activity that precisely mimics natural vocalizations was modulated by arginine-vasotocin, isotocin and their antagonists delivered to the preoptic area-anterior hypothalamus, a primary site for behavioural integration in all vertebrates. Peptides had different effects in males that acoustically court females (arginine-vasotocin-sensitive) than in females and sneak-spawning males (isotocin-sensitive), showing that the neuromodulatory mechanisms that establish reproduction-related behaviour can be dissociated from gonadal sex. PMID- 10693806 TI - Mapping the conformational wave of acetylcholine receptor channel gating. AB - Allosteric transitions allow fast regulation of protein function in living systems. Even though the end points of such conformational changes are known for many proteins, the characteristics of the paths connecting these states remain largely unexplored. Rate-equilibrium linear free-energy relationships (LFERs) provide information about such pathways by relating changes in the free energy of the transition state to those of the ground states upon systematic perturbation of the system. Here we present an LFER analysis of the gating reaction pathway of the muscle acetylcholine receptor. We studied the closed <==> open conformational change at the single-molecule level following perturbation by series of single site mutations, agonists and membrane voltages. This method provided a snapshot of several regions of the receptor at the transition state in terms of their approximate positions along the reaction coordinate, on a scale from 0 (closed like) to 1 (open-like). The resulting map reveals a spatial gradient of positional values, which suggests that the conformational change proceeds in a wave-like manner, with the low-to-high affinity change at the transmitter-binding sites preceding the complete opening of the pore. PMID- 10693807 TI - Positional cloning of zebrafish ferroportin1 identifies a conserved vertebrate iron exporter. AB - Defects in iron absorption and utilization lead to iron deficiency and overload disorders. Adult mammals absorb iron through the duodenum, whereas embryos obtain iron through placental transport. Iron uptake from the intestinal lumen through the apical surface of polarized duodenal enterocytes is mediated by the divalent metal transporter, DMTi. A second transporter has been postulated to export iron across the basolateral surface to the circulation. Here we have used positional cloning to identify the gene responsible for the hypochromic anaemia of the zebrafish mutant weissherbst. The gene, ferroportin1, encodes a multiple transmembrane domain protein, expressed in the yolk sac, that is a candidate for the elusive iron exporter. Zebrafish ferroportin1 is required for the transport of iron from maternally derived yolk stores to the circulation and functions as an iron exporter when expressed in Xenopus oocytes. Human Ferroportin1 is found at the basal surface of placental syncytiotrophoblasts, suggesting that it also transports iron from mother to embryo. Mammalian Ferroportin1 is expressed at the basolateral surface of duodenal enterocytes and could export cellular iron into the circulation. We propose that Ferroportin1 function may be perturbed in mammalian disorders of iron deficiency or overload. PMID- 10693808 TI - Interaction between Wnt and TGF-beta signalling pathways during formation of Spemann's organizer. AB - Members of the Wnt and TGF-beta superfamilies regulate both cell fate and proliferation during development and tissue maintenance. In the early amphibian embryo, the Wnt and TGF-beta superfamily signalling cascades are required for the establishment of a dorsal signalling centre, Spemann's organizer. Intracellular proteins of both pathways, upon activation, translocate to the nucleus to participate in transcription. Here we show that beta-catenin and Lef1/Tcf, which are downstream components of the Wnt signalling cascade, form a complex with Smad4, an essential mediator of signals initiated by members of the TGF-beta growth factor superfamily. In Xenopus, this interaction directly and synergistically affects expression of the twin (Xtwn) gene during formation of the organizer. This is, to our knowledge, the first demonstration of a physical interaction between TGF-beta and Wnt signalling components in vivo. PMID- 10693809 TI - Syncytin is a captive retroviral envelope protein involved in human placental morphogenesis. AB - Many mammalian viruses have acquired genes from their hosts during their evolution. The rationale for these acquisitions is usually quite clear: the captured genes are subverted to provide a selective advantage to the virus. Here we describe the opposite situation, where a viral gene has been sequestered to serve an important function in the physiology of a mammalian host. This gene, encoding a protein that we have called syncytin, is the envelope gene of a recently identified human endogenous defective retrovirus, HERV-W. We find that the major sites of syncytin expression are placental syncytiotrophoblasts, multinucleated cells that originate from fetal trophoblasts. We show that expression of recombinant syncytin in a wide variety of cell types induces the formation of giant syncytia, and that fusion of a human trophoblastic cell line expressing endogenous syncytin can be inhibited by an anti-syncytin antiserum. Our data indicate that syncytin may mediate placental cytotrophoblast fusion in vivo, and thus may be important in human placental morphogenesis. PMID- 10693810 TI - naked cuticle encodes an inducible antagonist of Wnt signalling. AB - During animal development, cells have to respond appropriately to localized secreted signals. Proper responses to Hedgehog, transforming growth factor-beta, epidermal growth factor and fibroblast growth factor/Ras signals require cognate inducible antagonists such as Patched, Dad, Argos and Sprouty. Wnt signals are crucial in development and neoplasia. Here we show that naked cuticle (nkd), a Drosophila segment-polarity gene, encodes an inducible antagonist for the Wnt signal Wingless (Wg). In fly embryos and imaginal discs nkd transcription is induced by Wg. In embryos, decreased nkd function has an effect similar to excess Wg; at later stages such a decrease appears to have no effect. Conversely, overproduction of Nkd in Drosophila and misexpression of Nkd in the vertebrate Xenopus laevis result in phenotypes resembling those of loss of Wg/Wnt function. nkd encodes a protein with a single EF hand (a calcium-binding motif) that is most similar to the recoverin family of myristoyl switch proteins. Nkd may therefore link ion fluxes to the regulation of the potency, duration or distribution of Wnt signals. Signal-inducible feedback antagonists such as nkd may limit the effects of Wnt proteins in development and disease. PMID- 10693811 TI - Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. AB - Yeast Sir2 is a heterochromatin component that silences transcription at silent mating loci, telomeres and the ribosomal DNA, and that also suppresses recombination in the rDNA and extends replicative life span. Mutational studies indicate that lysine 16 in the amino-terminal tail of histone H4 and lysines 9, 14 and 18 in H3 are critically important in silencing, whereas lysines 5, 8 and 12 of H4 have more redundant functions. Lysines 9 and 14 of histone H3 and lysines 5, 8 and 16 of H4 are acetylated in active chromatin and hypoacetylated in silenced chromatin, and overexpression of Sir2 promotes global deacetylation of histones, indicating that Sir2 may be a histone deacetylase. Deacetylation of lysine 16 of H4 is necessary for binding the silencing protein, Sir3. Here we show that yeast and mouse Sir2 proteins are nicotinamide adenine dinucleotide (NAD)-dependent histone deacetylases, which deacetylate lysines 9 and 14 of H3 and specifically lysine 16 of H4. Our analysis of two SIR2 mutations supports the idea that this deacetylase activity accounts for silencing, recombination suppression and extension of life span in vivo. These findings provide a molecular framework of NAD-dependent histone deacetylation that connects metabolism, genomic silencing and ageing in yeast and, perhaps, in higher eukaryotes. PMID- 10693812 TI - The structures of HsIU and the ATP-dependent protease HsIU-HsIV. AB - The degradation of cytoplasmic proteins is an ATP-dependent process. Substrates are targeted to a single soluble protease, the 26S proteasome, in eukaryotes and to a number of unrelated proteases in prokaryotes. A surprising link emerged with the discovery of the ATP-dependent protease HslVU (heat shock locus VU) in Escherichia coli. Its protease component HslV shares approximately 20% sequence similarity and a conserved fold with 20S proteasome beta-subunits. HslU is a member of the Hsp100 (Clp) family of ATPases. Here we report the crystal structures of free HslU and an 820,000 relative molecular mass complex of HslU and HslV-the first structure of a complete set of components of an ATP-dependent protease. HslV and HslU display sixfold symmetry, ruling out mechanisms of protease activation that require a symmetry mismatch between the two components. Instead, there is conformational flexibility and domain motion in HslU and a localized order-disorder transition in HslV. Individual subunits of HslU contain two globular domains in relative orientations that correlate with nucleotide bound and unbound states. They are surprisingly similar to their counterparts in N-ethylmaleimide-sensitive fusion protein, the prototype of an AAA-ATPase. A third, mostly alpha-helical domain in HslU mediates the contact with HslV and may be the structural equivalent of the amino-terminal domains in proteasomal AAA ATPases. PMID- 10693813 TI - Structure of the intact transactivation domain of the human papillomavirus E2 protein. AB - Papillomaviruses cause warts and proliferative lesions in skin and other epithelia. In a minority of papillomavirus types ('high risk, including human papillomaviruses 16, 18, 31, 33, 45 and 56), further transformation of the wart lesions can produce tumours. The papillomavirus E2 protein controls primary transcription and replication of the viral genome. Both activities are governed by a approximately 200 amino-acid amino-terminal module (E2NT) which is connected to a DNA-binding carboxy-terminal module by a flexible linker. Here we describe the crystal structure of the complete E2NT module from human papillomavirus 16. The E2NT module forms a dimer both in the crystal and in solution. Amino acids that are necessary for transactivation are located at the dimer interface, indicating that the dimer structure may be important in the interactions of E2NT with viral and cellular transcription factors. We propose that dimer formation may contribute to the stabilization of DNA loops which may serve to relocate distal DNA-binding transcription factors to the site of human papillomavirus transcription initiation. PMID- 10693814 TI - Knowing no fear. AB - People with brain injuries involving the amygdala are often poor at recognizing facial expressions of fear, but the extent to which this impairment compromises other signals of the emotion of fear has not been clearly established. We investigated N.M., a person with bilateral amygdala damage and a left thalamic lesion, who was impaired at recognizing fear from facial expressions. N.M. showed an equivalent deficit affecting fear recognition from body postures and emotional sounds. His deficit of fear recognition was not linked to evidence of any problem in recognizing anger (a common feature in other reports), but for his everyday experience of emotion N.M. reported reduced anger and fear compared with neurologically normal controls. These findings show a specific deficit compromising the recognition of the emotion of fear from a wide range of social signals, and suggest a possible relationship of this type of impairment with alterations of emotional experience. PMID- 10693815 TI - Anti-predator behaviour changes following an aggressive encounter in the lizard Tropidurus hispidus AB - Avoiding predators may conflict with territorial defence because a hiding territorial resident is unable to monitor its territory or defend it from conspecific intrusions. With persistent intruders, the presence of an intruder in the near past can indicate an increased probability of future intrusions. Therefore, following a conspecific-intrusion, territorial residents should minimize costs from future intrusions at the cost of higher predation risks. I conducted experiments with males of the territorial lizard Tropidurus hispidus recording approach distance (distance between predator and prey when the prey escapes) and time to re-emergence from a refuge after hiding. Past aggressive interactions affected anti-predator behaviour: lizards re-emerged sooner (compared to a control) when the predator attacked 5 min after an aggressive encounter. If the predator attacked while an aggressive encounter was ongoing, there was also a reduction in approach distance. The results are consistent with an economic hypothesis which predicts that T. hispidus incur greater predation risks to minimize future territorial intrusion; additionally they show that the effects of past and ongoing aggressive interactions are different, consistent with the minimization of present intrusion costs. These results are relevant for studies of the changes in aggressive behaviour due to changes in the social environment and for studies of the costs and (co) evolution of aggressive and anti-predator strategies. PMID- 10693816 TI - Regulation of queen-worker conflict in bumble-bee (Bombus terrestris) colonies AB - In annual colonies of bumble-bees overt queen-worker conflict is limited to a distinct 'competition phase' (CPh). In unmanipulated Bombus terrestris colonies, the queen's switch to male production (the 'switch point', SP) accounted for only 22% of the variation in the onset of the CPh. In some colonies, the CPh even began before the SP. The CPh was more strongly correlated with the transition in queen production (r=0.79). Replacing the queen eggs with male eggs or doubling the number of workers in young colonies resulted in a significantly earlier onset of the CPh and a significantly earlier transition to queen production. Replacing queen eggs with female eggs did not have this effect. These manipulations did not affect the timing of the queen's switch from female to male production. These findings show that the mechanism underlying the queen-worker conflict in insect societies is more complex than previously appreciated. The onset of queen-worker conflict cannot be attributed simply to a single factor such as the queen's switch to male production or a decrease in queen inhibition. Rather, multiple cues are important. PMID- 10693817 TI - An uncoupling of male and sexual egg production leads to reduced inbreeding in the cyclical parthenogen Daphnia AB - Cyclical parthenogenesis involves an alternation of parthenogenetic and sexual reproduction. In cyclical parthenogens with a short generation time, seasonal succession of clones switching to sexual reproduction at different periods of the growing season entails the risk of severe inbreeding with associated fitness costs. We show, however, that differences in genotype frequencies between males and sexual females result in a substantial reduction in the probability of selfing in natural Daphnia populations. This suggests that responses to male- and sexual egg-inducing stimuli may be uncoupled at the level of individual clones as a mechanism to avoid severe inbreeding. PMID- 10693818 TI - Sex differences in energy expenditure in non-human primates. AB - Female mammals bear the energetic costs of gestation and lactation. Therefore, it is often assumed that the overall energetic costs are greater for females than they are for males. However, the energetic costs to males of intrasex competition may also be considerable, particularly if males maintain a much larger body size than females. Using data from 19 non-human primates, this paper examines the relationship between male and female energetic costs both in the short term (daily energy expenditure) and the long term (the energetic cost of producing a single offspring). It is shown that the major determinant of sex differences in energetic costs is body size dimorphism. In the long term, the energetic costs are often greater for females, but, when male body size exceeds female body size by 60% or more, male energetic costs are greater than those for females. That is, in highly sexually dimorphic species the energetic costs of gestation and lactation for the females are matched by the energetic costs to the males of maintaining a large body size. PMID- 10693819 TI - Climate change has affected the breeding date of tree swallows throughout North America AB - Increasing evidence suggests that climate change has affected the breeding and distribution of wildlife. If such changes are due to global warming, then we should expect to see large-scale effects. To explore for such effects on avian reproduction, we examined 3450 nest records of tree swallows from across North America. The egg-laying date in tree swallows advanced by up to nine days during 1959-1991. This advance in phenology was associated with increasing surface air temperatures at the time of breeding. Our analysis controlled for several potentially confounding variables such as latitude, longitude, breeding density and elevation. We conclude that tree swallows across North America are breeding earlier and that the most likely cause is a long-term increase in spring temperature. PMID- 10693820 TI - Joint effects of feeding and breeding behaviour on trophic dimorphism in hummingbirds AB - A survey of 166 hummingbird species reveals novel associations of bill-length sexual dimorphism (BLSD) with plumage and breeding behaviours. Across all species, female bills become proportionately longer than male bills (higher female-to-male BLSD ratio) as sexual dichromatism increases. However, male bills are proportionately longer (lower female-to-male BLSD ratio) in both lekkers (traditional group display) and clustered breeders (female harems or colonial nests) compared with dispersed breeders. The overall positive association of plumage with BLSD suggests that social status determines priority of access to nectar-providing flowers. Furthermore, the distinctive BLSD associated with breeding aggregations may arise from behaviours that impose constraints on the usual male priority at flowers: female dominance over males around nest colonies and male residence on lek-mating territories. These various factors appear to alter plumage and bill characters of both sexes to produce the range of dimorphisms within the various dispersed and aggregated breeding system categories. Feedback loops caused by ecological consequences of breeding behaviour may alter the evolutionary dynamics of breeding systems, bird-plant interactions, and competing pollinators, as well as help explain the lek paradox. PMID- 10693821 TI - Bird orientation at high latitudes: flight routes between Siberia and North America across the Arctic Ocean AB - Bird migration and orientation at high latitudes are of special interest because of the difficulties associated with different compass systems in polar areas and because of the considerable differences between flight routes conforming to loxodromes (rhumblines) or orthodromes (great circle routes). Regular and widespread east-north-east migration of birds from the northern tundra of Siberia towards North America across the Arctic Ocean (without landmark influences) were recorded by ship-based tracking radar studies in July and August. Field observations indicated that waders, including species such as Phalaropusfulicarius and Calidris melanotos, dominated, but also terns and skuas may have been involved. Analysis of flight directions in relation to the wind showed that these movements are not caused by wind drift. Assuming possible orientation principles based on celestial or geomagnetic cues, different flight trajectories across the Arctic Ocean were calculated: geographical loxodromes, sun compass routes, magnetic loxodromes and magnetoclinic routes. The probabilities of these four alternatives are evaluated on the basis of both the availability of required orientation cues and the predicted flight paths. This evaluation supports orientation along sun compass routes. Because of the longitudinal time displacement sun compass routes show gradually changing compass courses in close agreement with orthodromes. It is suggested that an important migration link between Siberia and North American stopover sites 1000-2500km apart across the Arctic Ocean has evolved based on sun compass orientation along orthodrome-like routes. PMID- 10693822 TI - Evolution of stepping-stone dispersal rates. AB - We present a general model of the evolution of dispersal in a population with any distribution of dispersal distance. We use this model to analyse evolutionarily stable (ES) dispersal rates for the classical island model of dispersal and for three different stepping-stone models. Using general techniques to compute relatedness coefficients in the different dispersal models which we consider, we find that the distribution of dispersal distance may affect the ES dispersal rate when the cost of dispersal is low. In this case the ES dispersal rate increases with the number of demes that can be reached by one dispersal event. However, for increasing cost the ES dispersal rate converges to a value independent of the distribution of dispersal distance. These results are in contrast to previous analyses of similar models. The effects of the size (number of demes) and shape (ratio between the width and the length) of the population on the evolution of dispersal are also studied. We find that larger and more elongated populations lead generally to higher ES dispersal rates. However, both of these effects can only be observed for extreme parameter values (i.e. for very small and very elongated populations). The direct fitness method and the analytical techniques used here to compute relatedness coefficients provide an efficient way to analyse ES strategies in subdivided populations. PMID- 10693823 TI - The immunocompetence handicap hypothesis: testing the genetic predictions AB - The immunocompetence handicap hypothesis suggests that the immune system competes for resources with sexually selected ornaments; variation in ornaments might reflect genetic variation for immunocompetence. We tested this genetic prediction by mating scorpionfly females to males differing in the expression of a condition dependent ornament trait, saliva secretion, and then comparing offspring immunocompetence. We found several indications of an immunocompetence handicap in our study: females had superior immunocompetence compared with males, the different immune traits were positively correlated, and there were indications of genetic variation in immune traits. However, we found no significant difference in the immunocompetence of offspring derived from males differing in ornament expression, only a tendency for sons of ornamented males to possess slightly better immunocompetence. The estimated effect of fathers on offspring immunocompetence was rather small, but it might be a sufficient benefit of female choice, provided that the costs of choice are small. We conclude that the genetic benefit of female choice is small concerning offspring immunocompetence, but the immunocompetence handicap principle might nevertheless work in scorpionflies. PMID- 10693824 TI - Virus phenotype switching and disease progression in HIV-1 infection. AB - One of the phenotypic distinctions between different strains of human immunodeficiency virus type 1 (HIV-1) has to do with the ability to cause target cells to form large multinucleate bodies known as syncytia. There are two phenotypes according to this characterization: syncytium-inducing (SI) and non syncytium-inducing (NSI). NSI strains are usually present throughout infection, while SI strains are typically seen at the beginning of the infection and near the onset of AIDS. The late emergence of SI strains is referred to as phenotype switching. In this paper we analyse the factors that lead to phenotype switching and contribute to the dynamics of disease progression. We show that a strong immune system selects for NSI strains while a weak immune system favours SI strains. The model explicitly accounts for the fact that CD4+ cells are both targets of HIV infection and crucial for activating immune responses against HIV In such a model, SI strains can emerge after a long and variable period of NSI dominated infection. Furthermore, versions of the model which do not explicitly account for HIV-specific, activated CD4+ cells do not exhibit phenotype switching, emphasizing the critical importance of this pool of cells. PMID- 10693825 TI - Scrapie transmission in Britain: a recipe for a mathematical model. AB - Responses to an anonymous postal survey concerning scrapie are analysed. Risk factors associated with farms that have had scrapie are identified as size, geographical region, lambing practices and holding of certain breeds. Further analysis of farms that have scrapie only in bought-in animals reveals that such farms tend to breed a smaller proportion of their replacement animals than farms without scrapie. Farms that have had scrapie in home-bred animals have attributes associated with breeding many animals: large numbers of rams bought, few ewes bought, and many animals that are home-bred. The demography of British sheep farms as described by size, breeds, purchasing behaviour, age structure and proportion of animals that are home-bred is summarized. British farms with scrapie reveal certain special features: they have more sheep that are found dead, more elderly ewes and more cases of scab. PMID- 10693826 TI - Invasion thresholds for fungicide resistance: deterministic and stochastic analyses AB - Fungicide resistance is an important practical problem, but one that is poorly understood at the population level. Here we introduce a simple nonlinear model for fungicide resistance in botanical epidemics which includes the dynamics of the chemical control agent and the host population, while also allowing for demographic stochasticity in the host-parasite dynamics. This provides a mathematical framework for analysing the risk of fungicide resistance developing by including the parameters for the amount applied, longevity and application frequency of the fungicide. The model demonstrates the existence of thresholds for the invasion of the resistant strain in the parasite population which depend on two quantities: the relative fitness of the resistant strain and the effectiveness of control. This threshold marks a change from definite elimination of the resistant strain below the threshold to a finite probability of invasion which increases above the threshold. The fungicide decay rate, the amount of fungicide applied and the period between applications affect the effectiveness of control and, consequently, they influence whether or not resistance develops and the time taken to achieve a critical frequency of resistance. All three parameters are amenable to control by the grower or by coordinating the activity of a population of growers. Providing crude estimates of the effectiveness of control and relative fitness are available, the results can be used to predict the consequences of changing these parameters for the risk of invasion and the proportion of sites at which this might be expected to occur. Although motivated for fungicide resistance, the model has broader application to herbicide, antibiotic and antiviral resistance. The modelling approach and results are discussed in the context of resistance to chemical control in general. PMID- 10693827 TI - Validity of self-reported duration of work postures obtained by interview. MUSIC Norrtalje Study Group. AB - The aim of the study was to validate interview data concerning the duration of four work postures (1) sitting, (2) standing/walking with hands above shoulder level, (3) standing/walking with hands between shoulder and knuckle level, and (4) standing/walking with hands below knuckle level. The self-reported time spent in each posture was tested in relation to observations and technical measurements in 20 subjects during two full working days. The linear relationships between self-reports and observations were strong for the three postures; sitting (r2 = 0.55), hands above shoulder level (r2 = 0.58) and hands below knuckle level (r2 = 0.69). Thus, using this interview technique, self-reports concerning time spent in (1) sitting, (2) standing/walking with hands above shoulder level and, (3) standing/walking with hands below knuckle level may be accurate enough for studying these work postures in epidemiological studies. PMID- 10693828 TI - An ergonomic design and performance evaluation of pipettes. AB - This paper describes the results of an investigation of the differences in performance, postures, strains on hand-arm-shoulder musculature, and subjective ratings of three pipettes (models A, B, and C). Both models A and B were pipettes available on the market. Model C was developed for this study of an ergonomically designed pipette. The gripping posture of the three models was distinct both in the anatomical and in the functional sense. Working with models A and B required a four-finger grasp with a thumb operated plunger. Model C required a finger palmar power grip and the plunger was operated by the fingers. Performance evaluation of the different pipettes in different tasks indicated that using the proposed model C resulted in a 2-3% lower fault rate, a 10% shorter completion time, and the highest subjective ratings among the three. Postural analysis results indicated that when using model C, the shoulder was the least abducted, the wrist was the least extended, and the wrist was the least radially extended. Model C appeared to provide the greatest opportunity for delicate adjustments of posture in response to the activity of the skin receptors and reduced the strains on the upper body musculature, justifying the ergonomic input into the design. PMID- 10693829 TI - Assessment of workload and arm position during different work sequences: a study with portable devices on construction workers. AB - It is recognised that work related shoulder pain is overrepresented among construction workers compared to other occupations. Studies have shown that working with hands above shoulder level increases the shoulder load. Most studies have been confined to the laboratory. The present project was carried out to map the muscular engagement and postures of construction workers undertaking ceiling fitting, and to compare the results to those of the laboratory studies. Two ambulatory devices were used, one allowing recording of electromyographic (EMG) signals bilaterally from the trapezius muscle, and the other to record the position of both arms and back by means of measuring the angles between the vertical line and the back and both upper arms. These recordings were performed during 1.5-2 h work sequences. The results show that the work was mostly performed in an upright position, that both arms were used to a similar amount and that the workers for a large proportion of their working time had their upper arms at levels that are considered harmful in view of shoulder load. The EMG data showed that nearly 50% of the work was spent with trapezius activity that exceeded that of the reference contraction used (about 15% of maximal voluntary contraction) and that the time spent in muscular relaxation was 10%. It was concluded that the exposure of construction workers undertaking ceiling fitting meets the criteria formulated on the basis of laboratory experiments with respect to a high risk of acquiring chronic shoulder pain, due to rotator cuff tendinitis. PMID- 10693830 TI - The significance of lateral whole-body vibrations related to separately and simultaneously applied vertical motions. A validation study of ISO 2631. AB - Sensitivity of lateral motions relative to vertical motions were determined and compared to predictions provided by ISO 2631. Two experiments were executed where lateral and vertical motions were applied consecutively or simultaneously and where the magnitude of a single- or dual-axis test signal was adjusted until it was judged as equivalent to a preceding single-axis reference motion of the same frequency. Experiment 1: References consisted of vertical sinusoidal motions presented with 1.6-12.5 Hz and weighted accelerations of a(zw) = 0.3, 0.6 and 1.2 m s(-2) r.m.s., single-axis test signals were lateral motions of the same frequency. 26 subjects (15 men, 11 women, 20-56 yr) participated in the experiments. Accelerations adjusted for lateral vibrations above 1.6 Hz were considerably lower than predicted suggesting that the weighting factors provided in ISO 2631 are incorrect. Experiment 2: References consisted of single-axis vertical or lateral sinusoidal motions presented with 1.6-12.5 Hz and a weighted acceleration of a(zw) = 1.25 m s(-2) r.m.s. The dual-axis test signals consisted of a constant fraction of the reference acceleration (10, 25, 50, 75, 90%) and a perpendicularly oriented adjustable component. 31 subjects (15 men, 16 women, 19 51 yr) participated in the experiments. Both experiments revealed that ISO 2631 is qualitatively valid, the weighting of lateral motions above 1.6 Hz, however, should be increased in order to meet the actual sensitivity particularly in case of multi-axis vibrations. PMID- 10693831 TI - Maximum isometric trunk muscle strength and activity at trunk axial rotation during sitting. AB - This study was performed to provide information relating to the twisted posture being characteristic of the driver of an agricultural tractor working in the field. The relationship of trunk axial strength and muscle activity to trunk twisting angle of prerotation was determined and quantified. Differences between tractor drivers and office workers, and between the two directions of twisting action were also studied. Nine male tractor drivers and nine male office workers performed isometric maximum efforts at about -40, -20, 0, 20 and 40 degrees of pre-set trunk twisting angles in both the clockwise and counterclockwise directions. Exerted torque, true angle of prerotation and muscle activity from left and right side of each of obliquus externus, rectus abdominis and erector spinae were measured simultaneously. The results showed that the subjects could exert the greatest torques when being prerotated in the opposite direction and the lowest torques when being prerotated in the same direction to the direction of exertion. The exerted torques were within the range of 65-145 Nm. There were large differences in obliquus externus and erector spinae activity due to the twisting direction. There were also changes in muscle activity from obliquus externus and rectus abdominis due to prerotation angle. The results raised questions concerning the involvement of the passive tissues and the use of deeper muscles during trunk axial rotation, which should be further investigated. PMID- 10693832 TI - Wrist positions and movements as possible risk factors during machine milking. AB - High prevalence of hand and wrist symptoms has been found in females working with machine milking. Therefore the aim of this study was to quantify the positions and movements of the wrist during machine milking, and to compare tethering and loose-housing systems with respect to this. Biaxial electrogoniometers and data loggers were used for recording flexion and deviation angles of both the right and left wrists in 11 healthy milkers. For each individual 25 min of representative work was recorded in each system. High values of dorsiflexion and radial deviation were found, which might induce an increased risk of carpal tunnel syndrome. Moreover, the velocity and repetitiveness were close to those values described in repetitive work with a high risk of elbow and hand disorders in the fish-processing industry and giro-form data entry work. According to our findings, the load on the upper extremities has increased with respect to dorsiflexed hand position and repetitiveness when milking in the modern loose housing milking system. This is probably due to the change of the working position and/or the higher productivity (number of cows that milked per time unit) in the loose-housing system as compared to the old-fashioned tethering system. These negative effects on wrist positions and movements should be considered when building new milking systems. PMID- 10693833 TI - Applying ergonomics to Applied Ergonomics: using structured abstracts. AB - BACKGROUND: Previous research with structured abstracts (i.e. those that contain sub-headings such as this one) has indicated that structured abstracts are of a higher quality, contain more information, and are easier to search than are abstracts produced in the traditional manner. AIM: The aim of this article is to indicate how such structured abstracts might be appropriate for Applied Ergonomics. METHOD: Three abstracts taken from a recent issue of Applied Ergonomics were re-written in a structured form. This involved re-sequencing the information presented in the originals, and including additional information- particularly that of a quantitative kind--to meet the requirements of the sub headings. Measures of word length, information content, readability, and reader preferences were then made. RESULTS: The results showed that there were differences between the three pairs of abstracts on these various measures but that, overall, in line with previous research, the structured abstracts were longer, more informative and judged to be clearer by their readers. COMMENT: The findings support the author's view that structured abstracts are more effective than traditional ones. PMID- 10693834 TI - Development and evaluation of a microprocessor-based ergonomic dosimeter for evaluating carpentry tasks. AB - This portable and self-contained lightweight microprocessor based Ergonomic Dosimeter is designed to collect continuously postural angles of the torso and the upper arm in the sagittal plane and the number of kneeling activities. Up to 4 h of task performance data can be stored in a non-volatile memory of the dosimeter, which can then be downloaded to a lap-top computer. The portable dosimeter was tested for test-retest reliability, compared with posture data obtained with a computer-based video analysis system and evaluated at a carpenter's apprentices school and at a construction site. The dosimeter was shown to be suitable for collecting posture and kneeling data for a prolonged period at construction sites. PMID- 10693835 TI - The development and evaluation of an ergonomic glove. AB - The primary intent of this study was to determine if a hand glove could be designed on a criterion of selective protection. Force distribution patterns on the palmar side of hand were obtained from various studies to develop zones of hand that needed protection. A new design for gloves was developed based on the principle of selective protection, where protective material is introduced in varying levels over different parts of the glove, in order to provide protection where it is most needed, and at the same time preserve the desirable dexterity and strength capabilities of the barehand, optimizing the trade-off between protection and performance. Two pairs of prototype gloves incorporating different levels of protection were fabricated and tested using a battery of performance tests and an algometer test for pressure sensitivity. The test battery comprising four dexterity tasks and a maximal voluntary grip strength task was used to assess a number of glove conditions, including the two prototype gloves developed. The results indicate that the performance of the prototype gloves are comparable, and that the performance times for the double glove and the two prototype gloves tested were not significantly different. For the grip strength, the two prototype gloves were better than the double glove. The assembly task performance for the prototype II (laminar glove) was significantly lower than that of the other glove types tested. It appears that gloves of variable thickness can be developed to afford adequate protection at zones of most need. Glove manufacturers are recommended to use an ergonomic approach in the design of gloves. Such an approach, besides protecting the safety objective of gloves, could enhance productivity considerably. PMID- 10693836 TI - Ergonomics evaluation of a manually operated cassava chipping machine. AB - A manually operated machine for chipping cassava was evaluated. Six farmers took part in the study, with physiological, postural, and subjective measurements being taken. Using the machine resulted in drudgery and postural discomfort. Following an iterative design process and using appropriate anthropometric measurements, an improved, adjustable prototype was developed. This was tested with the six farmers and six novice users. It was found to reduce discomfort and physiological strain, allowed a faster work-rate (with novice users) and was preferred by all users. The study demonstrated how ergonomics can play an important role in reducing drudgery and improving user satisfaction in technology development and transfer in developing countries. PMID- 10693837 TI - Overlooked issues in Time to Heal essays. PMID- 10693839 TI - Behold the patient-safety genie. PMID- 10693838 TI - The relevance of community-oriented primary care for training preventive medicine residents. PMID- 10693840 TI - Collaborative care: a new model for a new century. AB - From the imagined vantage point of the year 2020, the author recounts the problems and deficiencies of health care in the 1990s and describes how academic medicine's leaders successfully confronted them. A key part of their strategy was to work together to form a coordinated network of medical schools, teaching hospitals, and academically oriented health systems, along with their staffs and a variety of community-based partners. In this way, they set a national agenda, pursued common goals, freely shared information and best practices, and cooperated to optimize their effectiveness in education, research, and clinical care. A major outcome of this new network was the Collaborative Care model of health care, based on the premise that a basic purpose of the health care system is to achieve measurable improvements in the health of individuals and communities in ways that are cost-conscious, quality-driven, evidence-based, and patient-, family-, and community-oriented. Academic institutions formed strong partnerships with many stakeholders (e.g., purchasers of health care services) to make the Collaborative Care approach work. In addition, there were several other important keys to Collaborative Care's success, such as the full integration of clinical research with clinical care and the restoration of trust in the health care enterprise. The author returns to reality and the 1990s. He challenges academic medicine to pursue the Collaborative Care vision, saying that "we should not accept without challenge what we know to be abominable just because it appears to be inevitable.... Our choice is to continue to struggle for survival as the environment around us gets harsher and harsher ... or to fix the environment" using the power of collaboration to unleash academic medicine's unlimited creativity and wisdom. PMID- 10693841 TI - Educating future physicians for Ontario: phase II. AB - In 1990, a collaborative project was launched to determine what the people of Ontario expect of their physicians and how the programs that prepare future physicians should be changed in response. The project, called Educating Future Physicians for Ontario (EFPO), brought together the five Ontario medical schools, the Council of Ontario Faculties of Medicine (COFM); a nonprofit, charitable organization, Associated Medical Services (AMS); and the Ontario Ministry of Health. The first phase ran for five years and was described in the November 1998 issue of Academic Medicine. After an external review, the project was continued for a second phase (EFPO II) for four more years until December 1998; that second phase is the topic of this article. EFPO II (1) focused more on residents' education; (2) emphasized four of the EFPO I-created physician roles in project activities; (3) maintained the province-wide, inter-institutional medical education framework of phase I, but fostered greater involvement of the seven sites (five medical schools and two regional health centers) in project activities; (4) stressed five project components (e.g., needs assessment and community partnerships) and worked for collaboration among components at all sites; (5) enhanced the original EFPO I Fellowship Program by adding residents and community fellows to the existing fellowships and by initiating leadership development activities, all of which bode well for the future leadership of medical education in Ontario. Students and residents played a vital role in EPFO II. Most of EFPO II's objectives were met, but the overall view of external reviewers was that the project was less successful than EFPO I. For example, the impact on clinical education, especially residency education, was less than anticipated. On the other hand, the project helped encourage the wide adoption of the eight physician roles that originated in EFPO I and advanced faculty development and assessment activities based on these roles. A third phase of EFPO concerning continuing medical education was planned, but support was not available. However, one of the funders will continue to support the successful fellowship and leadership program and the provincial education network for the next three years. Overall, the two phases of EFPO substantially modified medical education in Ontario to make it more responsive to evolving social needs. PMID- 10693842 TI - The ethics of caring and medical education. AB - The ethics of caring, though the subject of much recent discussion by philosophers, has hardly been applied to medical ethics and medical education. Based on receptivity (that is, empathy and compassion) toward and taking responsibility for other persons, the ethics of caring has particular relevance to medicine. Caring guides the physician always to remain the patient's advocate and to maintain the therapeutic relationship when dealing with and resolving ethical dilemmas. This article discusses the philosophy behind the ethics of caring and then explores three issues that arise within its context: receptivity, taking responsibility, and creating an educational environment that fosters caring. PMID- 10693843 TI - Programs for the development of physician leaders: a curricular process in its infancy. AB - Physician leaders are crucial as never before to ensure the proper integration of good care and cost containment; such integration is both a reasonable expectation of patients and essential for the survival of health care delivery systems. In today's health care environment, a critical mass of physician leaders must be developed in a systematic fashion so that physicians may truly lead the health care enterprise. The authors (1) describe, with examples, the various types and levels of physician leadership training programs currently being offered; (2) explain the costs and benefits of each program type; and (3) offer a program rationale and model (using a program at their medical school), which they analyze using traditional management concepts such as strategic planning, net present value, and make-versus-buy. The authors emphasize that physician leadership training should be local, offer long-term instruction, and be led by physicians. They conclude by stating that the concept of physician leadership will not and should not be taken seriously by non-physician health care executives until the physician community becomes as serious about leadership and management training as it is about clinical training. PMID- 10693845 TI - Defining educational objectives at the University of Virginia. PMID- 10693844 TI - The path to professionalism: cultivating humanistic values and attitudes in residency training. AB - Though few question the importance of incorporating professionalism and humanism in the training of physicians, traditional residency programs have given little direct attention to the processes by which professional and humanistic values, attitudes, and behaviors are cultivated. The authors discuss the underlying philosophy of their primary care internal medicine residency program, in which the development of professionalism and humanism is an explicit educational goal. They also describe the specific components of the program designed to create a learner-centered environment that supports the acquisition of professional values; these components include a communication-skills training program, challenging-case conferences, home visits with patients, a resident support group, and a mentoring program. The successful ten-year history of the program shows how a residency program can enable its trainees to develop not only the requisite excellent diagnostic and technical tools and skills but also the humane and professional attributes of the fully competent physician. PMID- 10693846 TI - The use of an automatic projector for a collateral film course in community health. PMID- 10693847 TI - Medicine and the arts. Girl, Interrupted. PMID- 10693848 TI - Specialty choices, compensation, and career satisfaction of underrepresented minority faculty in academic medicine. AB - PURPOSE: Despite efforts to increase the numbers of underrepresented minorities (URMs), only 3.9% of medical school faculty are URMs. The authors compared the specialty choices, compensation, and career satisfaction of minority faculty with those of their majority counterparts to determine whether there were differences that might affect the recruitment and retention of minority faculty. METHOD: In 1995, the authors mailed a self-administered survey to a stratified random sample of 3,013 eligible full-time salaried faculty in 24 randomly selected medical schools. Those schools, which had at least 200 faculty, did not include the Puerto Rican or historically black medical schools. RESULTS: Of the eligible faculty surveyed, 1,807 (60%) responded; 1,463 were majority faculty, 195 were URM faculty, and 149 were other-minority faculty. Similar proportions of the three groups were in the primary care specialties. Only 11% of the URM respondents were in basic science departments. There was no significant difference in adjusted mean compensation between majority, URM, and other minority faculty. However, URM faculty were significantly less satisfied with their careers (adjusted scores: 60 versus > 65; p = .001) and more often considered leaving academic medicine within five years (58% versus < 45%). CONCLUSION: Given the demographic changes of the U.S. population, these issues should be addressed by deans and department heads in order to enhance recruitment and facilitate retention of URM faculty in academic medicine. PMID- 10693849 TI - Developing and testing an instrument to measure the effectiveness of clinical teaching in an academic medical center. AB - PURPOSE: Instruments that rate teaching effectiveness provide both positive and negative feedback to clinician-educators, helping them improve their teaching. The authors developed the Clinical Teaching Effectiveness Instrument, which was theory-based and generic across their entire academic medical center, The Cleveland Clinic Foundation. They tested it for reliability, validity, and usability. METHOD: In 1997, using an iterative qualitative development process involving key stakeholders, the authors developed an institution-wide instrument to routinely evaluate clinical faculty. The resulting instrument has 15 questions that use a five-point evaluation scale. The instrument, which was administered to medical students, residents, and fellows over a 20-month period, produced data that were rigorously tested for instrument characteristics, reliability, criterion-related and content validity, and usability. RESULTS: This instrument, implemented in all departments across the institution, produced data on a total of 711 clinician-educators. Correlation coefficients among the items were high (.57 to .77). The scores were reliable (g coefficient of 0.935), and the instrument had both content and criterion-related validity. CONCLUSIONS: The Cleveland Clinic's Clinical Teaching Effectiveness Instrument is reliable and valid, as well as usable. It can be used as an evaluation tool for a wide variety of clinical teaching settings. PMID- 10693850 TI - Assessing how well three evaluation methods detect deficiencies in medical students' professionalism in two settings of an internal medicine clerkship. AB - PURPOSE: To compare the performances of three evaluation methods in detecting deficiencies of professionalism among third-year medical students during their ambulatory care and inpatient ward rotations of a core internal medicine clerkship. METHOD: From 1994 to 1997, 18 students at The Uniformed Services University of the Health Sciences failed to satisfactorily complete their core 12 week third-year internal medicine clerkship due to deficiencies in professionalism. Three evaluation methods had been used to assess all students' professionalism during the two rotations of their clerkship: standard checklists, written comments, and comments from formal evaluation sessions. Using qualitative methods and the information obtained by the three evaluation methods, the authors abstracted the record of each student concerning his or her clerkship behavior in terms of the six domains of professionalism used on the standard checklist. A detection index, which is the percentage of all instructors' less-than-acceptable ratings of a student across the six professionalism domains, was calculated for each evaluation method for each of the two clerkship settings. RESULTS: For each evaluation method, deficiencies in professionalism were twice as likely to be identified during the ward rotation as during the ambulatory care rotation (p < .002 for all). Formal evaluation session comments had the highest detection index in both clinical settings. Although the numbers of written and formal evaluation session comments per evaluator and per cited professionalism domain were similar, nearly a fourth of the instructors made identifying comments at the evaluation sessions only. CONCLUSION: In the clerkship studied, deficiencies in professionalism of such magnitude as to require remediation were more likely to be identified in the inpatient than in the ambulatory care setting. Of the three evaluation methods studied, the face-to-face, formal evaluation sessions significantly improved the detection of unprofessional behavior in both clerkship settings. Further efforts at such an interactive evaluation process with ambulatory care clerkship instructors may be essential for improving the identification of unprofessional behavior in that setting. PMID- 10693851 TI - Results of remedial continuing medical education in dyscompetent physicians. AB - PURPOSE: Noticing that moderately to severely incompetent physicians (as measured by a standardized assessment of physician competence) did not improve after traditional remedial continuing medical education (CME), the authors investigated the effects of a polyvalent, intensive, prolonged educational intervention on five physicians' competence. METHOD: The five physicians participated in a CME program that lasted three years and consisted of individualized review, ongoing small-group and evidence-based discussions, simulated patients and role playing, formal chart review, and peer review. At the end of the program, the physicians were reassessed. RESULTS: Only one physician improved; another remained the same, and three deteriorated. CONCLUSION: Successful remediation of severely incompetent physicians is uncertain at best, even with prolonged, intensive CME that incorporates modalities thought to be effective in changing physicians' behaviors. Alternative educational techniques may need to be developed for this select population. Conversely, there may be reasons that preclude improvement even with optimal techniques. PMID- 10693852 TI - Cognitive difficulty in physicians. AB - PURPOSE: Remediation of some incompetent physicians has proven difficult or impossible. The authors sought to determine whether physicians with impaired competency had neuropsychological impairment sufficient to explain their incompetence and their failure to improve with remedial continuing medical education (CME). METHOD: During a one-year period, 1996-97, all 27 participants in the Physician Review Program (PREP) conducted at McMaster University, a physician competency assessment program, undertook a detailed neuropsychological screening battery. RESULTS: Nearly all physicians assessed as competent also performed well on the neuropsychological testing. However, a significant number (about one third) of the physicians who performed poorly on the competency assessment had neuropsychological impairments sufficient to explain their poor performances. The difficulties were more marked in elderly physicians. CONCLUSION: A significant minority of incompetent physicians have cognitive impairments sufficient to explain both their incompetence and, probably, their failure to improve with remedial CME. Testing physicians for these impairments is important: to detect and treat reversible conditions, to manage irreversible conditions that preclude successful educational intervention, and to facilitate compensation in this instance. Serious consideration should be given to the incorporation of neuropsychological screening in all intensive physician review programs. PMID- 10693853 TI - Medicine and industry, medicine as industry. PMID- 10693854 TI - Scripts and medical diagnostic knowledge: theory and applications for clinical reasoning instruction and research. AB - Medical diagnosis is a categorization task that allows physicians to make predictions about features of clinical situations and to determine appropriate course of action. The script concept, which first arose in cognitive psychology, provides a theoretical framework to explain how medical diagnostic knowledge can be structured for diagnostic problem solving. The main characteristics of the script concept are pre-stored knowledge, values acceptable or not acceptable for each illness attribute, and default values. Scripts are networks of knowledge adapted to goals of clinical tasks. The authors describe how scripts are used in diagnostic tasks, how the script concept fits within the clinical reasoning literature, how it contrasts with competing theories of clinical reasoning, how educators can help students build and refine scripts, and how scripts can be used to assess clinical competence. PMID- 10693855 TI - Teaching internal medicine residents about medical problems in pregnancy. AB - When they became aware that many of their internal medicine residents were not being routinely exposed to a representative range of medical illnesses in pregnancy, the authors set out to develop and implement a brief practical curriculum on the medical problems of pregnancy. They began with a retrospective chart review of 562 consultations with pregnant women and used their findings to develop nine 15-minute lectures that covered a majority of the concepts essential to the care of the medically compromised pregnant woman. Topics included hypertension in pregnancy, the febrile pregnant woman, and renal disease in pregnancy. The authors also created a learner handout, a teaching script, teaching cases, and a bibliography for each lecture. Residents have responded well to the curriculum, and their mean pre- and posttext scores have shown that the lectures improved their knowledge of obstetric medicine. This brief-lecture format may be adapted to other special topics in residency training and readily integrated into already-crowded training schedules. PMID- 10693856 TI - The academic medical center of the future: a center for integrative study. AB - The integrative sciences, which are of great practical and conceptual interest, investigate complex systems (cells, multicellular organisms, families, institutions, communities, nations, etc.) at the higher levels of organization. Academic medical centers are intrinsically integrative because of their mission and thus are likely places for integrative study to flourish. They also possess vast resources of the kind needed to implement integrative studies. They can thus begin to address, from a scholarly point of view, a variety of integrative issues: how myriad parts (molecules, cells, organisms) form stable, complex, living wholes; the dynamics of health, illness, healing, dying, and death; problems of integration relating to patient care, health care delivery systems, and medical education. If academic medical centers undertake this mission they can be springboards for scholarly advances that will potentially affect all areas of thought. The authors describe how an integrative studies program can be started, and share the experiences of the University of Kentucky's Office of Integrative Studies. PMID- 10693857 TI - Outpatient morning report: a new educational venue. AB - Increasingly, medical educators are looking for ways to train residents and medical students in outpatient medicine. One novel idea, outpatient morning report, draws upon the concept of inpatient morning report and applies a similar conference format to the outpatient setting. The authors describe outpatient morning report and comment on its successful use in their institution. PMID- 10693858 TI - How to improve teaching in medical colleges. 1950. PMID- 10693859 TI - Measuring faculty effort and contributions in medical education. AB - A national panel on medical education was appointed as a component of the AAMC's Mission-based Management Program and charged with developing a metrics system for measuring medical school faculty effort and contributions to a school's education mission. The panel first defined important variables to be considered in creating such a system: the education programs in which medical school faculty participate; the categories of education work that may be performed in each program (teaching, development of education products, administration and service, and scholarship in education); and the array of specific education activities that faculty could perform in each of these work areas. The panel based the system on a relative value scale, since this approach does not equate faculty performance solely to the time expended by a faculty member in pursuit of a specific activity. Also, a four-step process to create relative value units (RVUs) for education activities was developed. This process incorporates quantitative and qualitative measures of faculty activity and also can measure and value the distribution of faculty effort relative to a school's education mission. When adapted to the education mission and culture of an individual school, the proposed metrics system can provide critical information that will assist the school's leadership in evaluating and rewarding faculty performance in education and will support a mission-based management strategy in the school. PMID- 10693861 TI - The future status of pediatric rheumatology in the United States: strategic planning for the year 2000. American College of Rheumatology Blue Ribbon Committee for Academic Pediatric Rheumatology. PMID- 10693862 TI - Approaches for identifying and defining environmentally associated rheumatic disorders. PMID- 10693863 TI - Synovial tissue in rheumatoid arthritis is a source of osteoclast differentiation factor. AB - OBJECTIVE: Osteoclast differentiation factor (ODF; also known as osteoprotegerin ligand, receptor activator of nuclear factor kappaB ligand, and tumor necrosis factor-related activation-induced cytokine) is a recently described cytokine known to be critical in inducing the differentiation of cells of the monocyte/macrophage lineage into osteoclasts. The role of osteoclasts in bone erosion in rheumatoid arthritis (RA) has been demonstrated, but the exact mechanisms involved in the formation and activation of osteoclasts in RA are not known. These studies address the potential role of ODF and the bone and marrow microenvironment in the pathogenesis of osteoclast-mediated bone erosion in RA. METHODS: Tissue sections from the bone-pannus interface at sites of bone erosion were examined for the presence of osteoclast precursors by the colocalization of messenger RNA (mRNA) for tartrate-resistant acid phosphatase (TRAP) and cathepsin K in mononuclear cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to identify mRNA for ODF in synovial tissues, adherent synovial fibroblasts, and activated T lymphocytes derived from patients with RA. RESULTS: Multinucleated cells expressing both TRAP and cathepsin K mRNA were identified in bone resorption lacunae in areas of pannus invasion into bone in RA patients. In addition, mononuclear cells expressing both TRAP and cathepsin K mRNA (preosteoclasts) were identified in bone marrow in and adjacent to areas of pannus invasion in RA erosions. ODF mRNA was detected by RT-PCR in whole synovial tissues from patients with RA but not in normal synovial tissues. In addition, ODF mRNA was detected in cultured adherent synovial fibroblasts and in activated T lymphocytes derived from RA synovial tissue, which were expanded by exposure to anti-CD3. CONCLUSION: TRAP-positive, cathepsin K-positive osteoclast precursor cells are identified in areas of pannus invasion into bone in RA. ODF is expressed by both synovial fibroblasts and by activated T lymphocytes derived from synovial tissues from patients with RA. These synovial cells may contribute directly to the expansion of osteoclast precursors and to the formation and activation of osteoclasts at sites of bone erosion in RA. PMID- 10693864 TI - Involvement of receptor activator of nuclear factor kappaB ligand/osteoclast differentiation factor in osteoclastogenesis from synoviocytes in rheumatoid arthritis. AB - OBJECTIVE: To clarify the mechanism by which osteoclasts are formed in culture of rheumatoid synoviocytes by exploring the involvement of receptor activator of nuclear factor kappaB ligand (RANKL)/osteoclast differentiation factor (ODF). METHODS: Osteoclast formation was evaluated in cocultures of rheumatoid synovial fibroblasts and peripheral blood mononuclear cells (PBMC) in the presence of macrophage colony stimulating factor and 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) utilizing separating membrane filters. RANKL/ODF expression was examined by Northern blotting in synovial tissues from 5 rheumatoid arthritis (RA) patients and tissues from patients with giant cell tumor (GCT), osteosarcoma (OS), and osteoarthritis (OA). RANKL/ODF expression and the ability of synovial fibroblasts to support osteoclastogenesis were investigated in coculture with PBMC in the presence or absence of 1,25(OH)2D3, and soluble RANKL/ODF and osteoprotegerin (OPG)/osteoclastogenesis inhibitory factor (OCIF) were measured by enzyme-linked immunosorbent assay. The effects of OPG/OCIF on the osteoclastogenesis in the primary culture of rheumatoid synoviocytes and the coculture system were determined. RESULTS: Synovial fibroblasts did not induce osteoclastogenesis when separately cocultured with PBMC. Northern blotting revealed that RANKL/ODF was highly expressed in all tissues from RA and GCT patients, but not from OA or OS patients. Cultured rheumatoid synovial fibroblasts efficiently induced osteoclastogenesis in the presence of 1,25(OH)2D3, which was accompanied by up regulated expression of RANKL/ODF and decreased production of OPG/OCIF. Osteoclastogenesis from synoviocytes was dose-dependently inhibited by OPG/OCIF. CONCLUSION: RANKL/ODF expressed on synovial fibroblasts is involved in rheumatoid bone destruction by inducing osteoclastogenesis and would therefore be a good therapeutic target. PMID- 10693865 TI - The effects of interferon-beta treatment of synovial inflammation and expression of metalloproteinases in patients with rheumatoid arthritis. AB - OBJECTIVE: Interferon-beta (IFNbeta) treatment is emerging as a potentially effective form of therapy in various immune-mediated conditions. This study evaluated the effects of IFNbeta therapy on the cell infiltrate, cytokine profile, and expression of metalloproteinase 1 (MMP-1) in synovial tissue from patients with rheumatoid arthritis (RA). To further assess the mechanism of action, in vitro experiments were conducted to determine the effects of IFNbeta on the production of MMP-1, MMP-3, tissue inhibitor of metalloproteinases 1 (TIMP 1), and prostaglandin E2 (PGE2) by human fibroblast-like synoviocytes (FLS). METHODS: Eleven patients were treated for 12 weeks with purified natural fibroblast IFNbeta (Frone; Ares-Serono, Geneva, Switzerland) subcutaneously 3 times weekly with the following dosages: 6 million IU (n = 4), 12 million IU (n = 3), and 18 million IU (n = 4). Synovial biopsy specimens were obtained by needle arthroscopy at 3 time points: directly before and at 1 month and 3 months after initiation of treatment. Immunohistologic analysis was performed using monoclonal antibodies specific for the following phenotypic markers and mediators of joint inflammation and destruction: CD3, CD38, CD68, CD55, tumor necrosis factor alpha (TNFalpha), interleukin-1beta (IL-1beta), IL-6, MMP-1, and TIMP-1. In addition, we measured the production of MMP-1, MMP-3, TIMP-1, and PGE2 by RA FLS and dermal fibroblasts in the presence and absence of IFNbeta. RESULTS: A statistically significant reduction in the mean immunohistologic scores for CD3+ T cells and the expression of MMP-1 and TIMP-1 at 1 month, CD38+ plasma cells and the expression of IL-6 at 3 months, and the expression of IL-1beta at both 1 and 3 months was observed in synovial tissue after IFNbeta treatment. The scores for CD68+ macrophages and TNFalpha expression also tended to decrease, but the differences did not reach statistical significance. The in vitro experiments revealed inhibition of MMP-1, MMP-3, and PGE2 production by RA FLS, whereas TIMP 1 production was only slightly decreased. These effects were more consistent in RA FLS than in dermal fibroblasts. CONCLUSION: The changes in synovial tissue after IFNbeta treatment and the in vitro data support the view that IFNbeta therapy has immunomodulating effects on rheumatoid synovium and might help to diminish both joint inflammation and destruction. Larger well-controlled studies are warranted to show the efficacy of IFNbeta treatment for RA. PMID- 10693866 TI - Increased expression of extracellular matrix metalloproteinase inducer in rheumatoid synovium. AB - OBJECTIVE: To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in the synovial membrane of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Mouse monoclonal antibody against human EMMPRIN was applied according to an avidin-biotin peroxidase complex method to reveal EMMPRIN expression. Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to check for the presence of EMMPRIN protein and messenger RNA (mRNA). RESULTS: EMMPRIN immunoreactivity was more intense in RA than in OA synovial membrane (P < 0.01). EMMPRIN staining was more widespread in RA than in OA, especially in association with macrophage infiltrates. RT-PCR of synovial membrane samples disclosed the presence of EMMPRIN mRNA. Nucleotide sequencing of the PCR amplification products confirmed the identity of the amplified bands. Immunoblot analysis revealed 55-kd glycosylated EMMPRIN bands, which were particularly prominent in RA samples. CONCLUSION: The expression of EMMPRIN is upregulated in the rheumatoid synovial membrane. EMMPRIN can induce local production of at least MMPs 1, 2, and 3, and can thereby play a role in joint destruction in RA. PMID- 10693867 TI - Synovial fluid levels of tumor necrosis factor alpha and oncostatin M correlate with levels of markers of the degradation of crosslinked collagen and cartilage aggrecan in rheumatoid arthritis but not in osteoarthritis. AB - OBJECTIVE: To compare synovial fluid (SF) levels of oncostatin M (OSM), tumor necrosis factor alpha (TNFalpha), and interleukin-6 (IL-6) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to determine which correlate best with SF levels of antigenic keratan sulfate (Ag KS), a marker of aggrecan catabolism, and pyridinium crosslinks, markers of the degradation of mature collagen molecules. METHODS: SF was drawn from the knee joints of patients with RA (n = 31) or OA (n = 31). Levels of Ag KS, D-pyridinoline (D-Pyr), pyridinoline (Pyr), OSM, TNFalpha, and IL-6 were measured by enzyme-linked immunosorbent assay. RESULTS: RA patients had higher median SF levels of OSM, TNFalpha, IL-6, and Pyr, but a lower median level of D-Pyr, than OA patients. In both groups, IL-6 levels correlated positively with those of OSM and TNFalpha. However, the correlation between levels of OSM and TNFalpha was only significant in the RA group. Ag KS and Pyr levels correlated positively in RA but not in OA. The correlation between TNFalpha and Ag KS was positive in RA and negative in OA. Further, in RA, OSM and IL-6 levels correlated strongly with Pyr and Ag KS levels but not with D-Pyr levels, while there were no strong correlations in OA for OSM or IL-6 levels with Pyr, Ag Ks, or D-Pyr levels. CONCLUSION: This in vivo study suggests that TNFalpha and other proinflammatory cytokines are involved in the up regulation of the coordinated degradation of cartilage aggrecan and collagen in RA. Further, OSM may act synergistically with other proinflammatory cytokines in up-regulating the production of metalloproteinases by chondrocytes in rheumatoid joints. PMID- 10693868 TI - Therapy and prevention of arthritis by recombinant adeno-associated virus vector with delivery of interleukin-1 receptor antagonist. AB - OBJECTIVE: To evaluate the recombinant adeno-associated virus vector encoding interleukin-1 receptor antagonist (rAAV-IL-1Ra) complementary DNA for its potential in the treatment and prevention of lipopolysaccharide (LPS)-induced arthritis. METHODS: The therapeutic effect of rAAV-IL-1Ra on arthritis was studied by injecting knees of Sprague-Dawley rats with LPS and rAAV-IL-1Ra and then evaluating the severity of arthritis by leukocyte counts in synovial fluid, histologic changes of synovium, and uptake of 67Ga citrate in joint tissue. To study the therapeutic effect on recurrent arthritis, we induced recurrent arthritis by a second injection of LPS 80 days after primary LPS and rAAV-IL-1Ra injections and then evaluated the severity of recurrent arthritis. To study the prevention of arthritis, rAAV-IL-1Ra was injected into normal joints. After 100 days, LPS was used to induce arthritis, and the severity of arthritis was evaluated. RESULTS: The production of the rAAV-IL-1Ra transgene was up-regulated by LPS-induced joint inflammation and proved to be efficacious in the therapeutic and preventative protocols. Not only primary but also recurrent arthritis could be suppressed by a single injection of rAAV-IL-1Ra. We found that the transgene expression of IL-1Ra could be reactivated by a second challenge with LPS delayed for 80 days after rAAV administration. The therapeutic level of IL-1Ra protein reached a mean +/- SD of 5.8+/-0.5 ng/ml in synovial fluid. In addition, the rAAV transgene persisted within normal joints for at least 100 days and could still be induced to express, after LPS insult, a high level of IL-1Ra (mean +/- SD 5.2+/ 0.8 ng/ml) that prevented the occurrence of arthritis. CONCLUSION: This gene therapy, by combining highly efficient and stable rAAV gene delivery, disease regulated gene expression, and the antiinflammatory effect of IL-1Ra, provides a valuable approach for long-term protection against, and prevention of, arthritis. PMID- 10693869 TI - 123I-antileukoproteinase scintigraphy reveals microscopic cartilage alterations in the contralateral knee joint of rats with "monarticular" antigen-induced arthritis. AB - OBJECTIVE: To assess the involvement of the contralateral knee joint in monarticular antigen-induced arthritis (AIA) by scintigraphy with the cationic (pI >10), 123I-labeled, serine proteinase inhibitor antileukoproteinase (123I ALP) and to compare the scintigraphic findings with those of radiography and high resolution ex vivo magnetic resonance imaging (MRI). METHODS: Lewis rats with chronic AIA were examined 2.5 months following arthritis induction (injection of 500 microg of methylated bovine serum albumin/saline into the ipsilateral [arthritic] knee joint and injection of phosphate buffered saline into the contralateral knee joint following systemic immunization). 123I-ALP was injected intravenously into normal rats (n = 4) or rats with AIA (n = 6). The ipsilateral and contralateral knee joints and both ankles were examined by scintigraphy and radiography. Joint cartilage was examined by high-resolution ex vivo MRI, histopathology, and measurement of tissue radioactivity. RESULTS: ALP accumulation (typically observed in normal articular cartilage) was lost in both the ipsilateral and the contralateral knee joints, but not in the clinically unaffected ankles of rats with AIA. In both knee joints, 123I-ALP target:background ratios and cartilage radioactivity correlated negatively with the loss of toluidine blue staining in cartilage, which documents the depletion of charged matrix molecules. Findings of histopathology confirmed mild alterations in the ipsilateral knee joint and even milder alterations in the contralateral knee joint, while the ankles were normal. Radiography and high resolution ex vivo MRI failed to detect abnormalities in the contralateral knee joint. CONCLUSION: Loss of ALP accumulation appears to document proteoglycan depletion, even in the microscopically altered cartilage of the contralateral knee joint in AIA. These findings underscore the high sensitivity of 123I-ALP for in vivo detection of biochemical cartilage alterations in arthritis, and furthermore, question the use of the contralateral knee joint as a normal control in AIA. PMID- 10693870 TI - Cyclosporin A prevents the histologic damage of antigen arthritis without inducing fibrosis. AB - OBJECTIVE: To study the effects of cyclosporin A (CSA) in a model of rheumatoid arthritis (RA) and the activation of growth factors related to pannus development at the site of injury. METHODS: Antigen arthritis was induced in the knees of 14 New Zealand rabbits, and the animals were randomized into 2 therapeutic groups: CSA at 10 mg/kg/day and CSA solvent. After 3 weeks of treatment, the rabbits were killed, and synovial tissues were obtained and compared with healthy specimens with regard to histopathologic lesions, deposition of transforming growth factor beta (TGFbeta) and collagens, and messenger RNA expression of platelet-derived growth factor B (PDGF-B) and TGFbeta. The effect of CSA on the expression of TGFbeta and PDGF-B was also examined in cultured synovial cells. RESULTS: CSA administration alleviated the histologic damage and avoided the overdeposition of matrix elements in the injured tissue. It was also able to normalize the enhanced expression of TGFbeta and PDGF-B observed in the untreated rabbits. Despite this modulation found in vivo, CSA up-regulated in a dose-dependent manner the gene expression of both trophic factors by healthy cultured synovial cells. CONCLUSION: The present study shows that continuous administration of CSA prevents the development of chronic synovitis in an experimental model of RA. As reported in other cell types, CSA promoted TGFbeta transcription by synovial cells in vitro, but failed to display a profibrogenic effect in the inflamed environment. PMID- 10693871 TI - Beneficial effects of tempol, a membrane-permeable radical scavenger, in a rodent model of collagen-induced arthritis. AB - OBJECTIVE: To investigate the effects of tempol, a membrane-permeable radical scavenger, in rats with collagen-induced arthritis (CIA). METHODS: CIA was induced in Lewis rats by intradermal injection of 100 microl of an emulsion of 100 microg of bovine type II collagen (CII) in complete Freund's adjuvant (FCA) at the base of the tail. On day 21, a second injection of CII in FCA was administered. RESULTS: Lewis rats developed an erosive arthritis of the hind paws when immunized with CII in FCA. Macroscopic evidence of CIA first appeared as periarticular erythema and edema in the hind paws. The incidence of CIA was 100% by day 27 in the CII-challenged rats, and the severity of CIA progressed over a 35-day period. Radiographs revealed focal resorption of bone, with osteophyte formation in the tibiotarsal joint, and soft tissue swelling. The histopathologic features included erosion of the cartilage at the joint margins. Treatment of rats with tempol (10 mg/kg/day intraperitoneally) starting at the onset of arthritis (day 23) delayed the development of the clinical signs on days 24-35 and improved the histologic status of the knee and paw. Immunohistochemical analysis for nitrotyrosine and poly(ADP-ribose) synthetase (PARS) revealed positive staining in the inflamed joints of CII-treated rats. The degree of nitrotyrosine and PARS staining was markedly reduced in tissue sections obtained from CII-treated rats that had received tempol. Furthermore, radiographs revealed protection against bone resorption and osteophyte formation in the joints of tempol-treated rats. CONCLUSION: This study is the first to provide evidence that tempol, a small molecule that permeates biologic membranes and scavenges reactive oxygen species, attenuates the degree of chronic inflammation and tissue damage associated with CIA in the rat. PMID- 10693872 TI - Evidence for the expression of a second CD6 ligand by synovial fibroblasts. AB - OBJECTIVE: CD6, a cell surface glycoprotein expressed primarily on T cells, may function as a costimulatory molecule and may play a role in autoreactive immune responses. Recently, a CD6 ligand termed CD166 (previously known as activated leukocyte cell adhesion molecule [ALCAM]) has been identified and shown to be expressed on activated T cells, B cells, thymic epithelium, keratinocytes, and in rheumatoid arthritis synovial tissue. However, the results of functional studies have suggested the existence of a second CD6 ligand. The present study was undertaken to seek evidence for a second CD6 ligand on cultured synovial fibroblasts. METHODS: Flow cytometric and biochemical techniques were applied, using anti-CD166 monoclonal antibody (mAb) and a recombinant CD6 fusion protein, to determine whether cultured synovial fibroblasts and other cell types expressed a non-ALCAM CD6 ligand. RESULTS: CD14- fibroblastic synoviocytes showed greater binding of a recombinant CD6 fusion protein than of anti-ALCAM mAb. With interferon-gamma treatment of synovial fibroblasts, binding of both reagents increased, but this was more marked for binding of CD6 fusion protein. Exposure of synovial fibroblasts to other cytokines or to the superantigen staphylococcal enterotoxin A also regulated binding of CD6 fusion protein and anti-ALCAM mAb in a discordant manner. Immunoprecipitation of proteins from membrane extracts of synovial fibroblasts with a CD6-Ig fusion protein revealed a novel 130-kd band distinct from CD166; an identical molecule was also precipitated from membranes of HBL-100 tumor cells. CONCLUSION: Taken together with previous data regarding CD6 and CD166 function, the present findings strongly suggest the existence of a second CD6 ligand distinct from CD166, which can be expressed by synovial fibroblasts as well as other cells. PMID- 10693873 TI - Decreased expression of interleukin-1alpha, interleukin-1beta, and cell adhesion molecules in muscle tissue following corticosteroid treatment in patients with polymyositis and dermatomyositis. AB - OBJECTIVE: To study the effects of immunosuppressive therapy, in particular, corticosteroids, on morphologic signs of inflammation and expression of cytokines, adhesion molecules, and class I major histocompatibility complex (MHC) antigen in muscle tissue from patients with polymyositis (PM) and dermatomyositis (DM) and to correlate the molecular changes with changes in muscle function. METHODS: Seven patients with PM and 4 patients with DM underwent muscle biopsy before and after 3-6 months of therapy. Ten of the 11 patients were initially treated with prednisolone 30-60 mg/day. The phenotypes of infiltrating inflammatory cells and the expression of interleukin-1alpha (IL-1alpha) and IL 1beta, adhesion molecules, and class I MHC antigen were studied by immunochemistry. Computerized image analysis was used for quantitation of staining. Muscle function was assessed with a muscle function index score. RESULTS: Pronounced improvement of muscle function during the treatment period was noted in 8 of the 11 patients. The changes in muscle function coincided with an almost complete disappearance of inflammatory cells, including CD3+ T cells, in the patients with clinical improvement. These patients also exhibited decreased expression of IL-1alpha, IL-1beta, intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), leukocyte function associated antigen 1alpha, and very late activation antigen 4alpha. Of note, there was persistent expression of IL-1alpha, ICAM-1, and VCAM-1 in capillaries and of class I MHC antigens on muscle fibers in several of the patients who, after corticosteroid treatment, still had muscle weakness despite the disappearance of inflammatory infiltrates. CONCLUSION: Changes in the muscle expression of key molecules in the inflammatory process, such as IL-1alpha and IL 1beta, ICAM-1 and class I MHC antigens, showed a consistent but not complete concordance with changes in and status of muscle function in patients with myositis who received the current standard treatment for the disease. These data indicate that it is possible to further evaluate various therapies for myositis using molecular analysis of muscle biopsy specimens obtained on repeated occasions. In addition, the data demonstrate a dissociation between muscle function and degree of inflammatory infiltration in the affected muscles and suggest that the functional defects are more related to the expression of molecules such as IL-1alpha in muscle capillaries than to the mere presence of inflammatory cells in the affected muscles. PMID- 10693874 TI - Identification of intervals on chromosomes 1, 3, and 13 linked to the development of lupus in BXSB mice. AB - OBJECTIVE: To identify intervals containing systemic lupus erythematosus (SLE) susceptibility alleles in the BXSB strain of mice. METHODS: We analyzed 286 (B10 x [B10 x BXSB]F1) backcross mice for a range of phenotypic traits associated with the development of SLE in BXSB mice. The mice were genotyped using 93 microsatellite markers, and the linkage of these markers to disease was studied by extreme-phenotype and quantitative trait locus analysis. RESULTS: The disease phenotype in these backcross mice was less severe than that in BXSB mice. However, antinuclear antibody production was increased compared with the parental strain. We identified 4 areas of genetic linkage to disease on chromosome 1 (Bxs1 4), 1 on chromosome 3 (Bxs5), and another interval on chromosome 13 which were associated with various aspects of the phenotype. Bxs4 and Bxs5 are located in regions not previously linked to disease in other models of SLE. CONCLUSION: SLE in the BXSB mouse model has a complex genetic basis and involves at least 5 distinct intervals located on chromosomes 1 and 3. There is evidence that different intervals affect particular aspects of the SLE phenotype. PMID- 10693875 TI - The features of arthritis induced by CpG motifs in bacterial DNA. AB - OBJECTIVE: To investigate the features of arthritis induced by bacterial DNA that contain CpG motifs. METHODS: Bacterial DNA originating from Escherichia coli and Staphylococcus aureus or synthetic oligonucleotides containing CpG motifs were injected directly into knee joints of mice. Histopathologic joint damage, antibody levels, cytokine levels, and synovial messenger RNA (mRNA) expression of cytokines and chemokines were assessed. RESULTS: Histopathologic signs of arthritis were evident within 2 hours and lasted for at least 3 weeks. Nonmethylated CpG motifs were responsible for the induction of arthritis since oligonucleotides containing these motifs triggered arthritis, whereas methylation of these nucleotides abrogated the inflammatory response. Arthritis was characterized by an influx of monocytic, Mac-1+ cells and by a scarcity of T lymphocytes. The disease was characterized locally by mRNA expression of macrophage-derived cytokines (tumor necrosis factor alpha, interleukin-12 [IL 12], IL-1beta) and chemokines (monocyte chemoattractant protein 1, RANTES) in arthritic joints. Systemically, the arthritis was characterized by increased levels of circulating IL-6 and immunoglobulins. CONCLUSION: These findings demonstrate that bacterial DNA that contain nonmethylated CpG motifs induces arthritis, suggesting an important pathogenic role for bacterial DNA in septic arthritis. PMID- 10693876 TI - Detection of viral ribonucleic acid and histologic analysis of inflamed synovium in Ross River virus infection. AB - OBJECTIVE: To document the histology of Ross River virus (RRV) arthritis and to examine inflamed synovium for viral RNA. METHODS: Biopsy tissue from the inflamed knees of 12 patients with RRV infection was studied using conventional and immunostaining techniques. Reverse transcriptase-polymerase chain reaction technology was used to probe for the presence of viral RNA in the synovial biopsy samples and in serum. RESULTS: Hyperplasia of the synovial lining layer, vascular proliferation, and mononuclear cell infiltration were the main histologic changes. RRV RNA was found in knee biopsy tissue that was obtained from 2 patients at 5 weeks after the onset of symptoms. CONCLUSION: RRV RNA was identified in inflamed synovium more than a month after symptoms began. Inflammation was apparent in the absence of detectable virus in the majority of patients. PMID- 10693877 TI - Comparison of the effect of rofecoxib (a cyclooxygenase 2 inhibitor), ibuprofen, and placebo on the gastroduodenal mucosa of patients with osteoarthritis: a randomized, double-blind, placebo-controlled trial. The Rofecoxib Osteoarthritis Endoscopy Multinational Study Group. AB - OBJECTIVE: This randomized, double-blind study tested the hypothesis that rofecoxib, a drug that specifically inhibits cyclooxygenase 2, would cause fewer gastroduodenal ulcers than ibuprofen (in a multicenter trial), and its side effects would be equivalent to those of placebo (in a prespecified analysis combining the results with another trial of identical design). METHODS: Seven hundred seventy-five patients with osteoarthritis were randomized to receive rofecoxib at a dosage of 25 mg or 50 mg once daily, ibuprofen 800 mg 3 times daily, or placebo. Gastroduodenal ulceration was assessed by endoscopy at 6, 12, and (for active treatment) 24 weeks. The primary and secondary end points were the incidence of gastroduodenal ulcers at 12 and 24 weeks, respectively. RESULTS: Ulcers were significantly less common (P < 0.001) following treatment with rofecoxib (25 mg or 50 mg) than with ibuprofen after 12 weeks (5.3% and 8.8% versus 29.2%, respectively) or 24 weeks (9.9% and 12.4% versus 46.8%, respectively). In the combined analysis, the 12-week ulcer incidence with 25 mg rofecoxib (4.7%) and with placebo (7.3%) satisfied prespecified criteria for equivalence. CONCLUSION: At 2-4 times the therapeutically effective dose, rofecoxib caused fewer endoscopically detected ulcers than did ibuprofen. Rofecoxib at a dose of 25 mg (the highest dose recommended for osteoarthritis) satisfied prespecified criteria for equivalence to placebo. PMID- 10693878 TI - Preference for nonsteroidal antiinflammatory drugs over acetaminophen by rheumatic disease patients: a survey of 1,799 patients with osteoarthritis, rheumatoid arthritis, and fibromyalgia. AB - OBJECTIVE: Because there is controversy regarding the efficacy of acetaminophen in rheumatic diseases and because apparently safer nonsteroidal antiinflammatory drugs (NSAIDs) are being produced, we surveyed rheumatic disease patients about their preferences for these agents to determine the degree to which one type of therapeutic agent is preferred over the other. METHODS: In 1998, we surveyed by mailed questionnaire 1,799 patients with osteoarthritis (OA), rheumatoid arthritis, or fibromyalgia who were participating in a long-term outcome study. Patients who had taken acetaminophen rated the effectiveness of acetaminophen, compared its effectiveness with that of NSAIDs, and then rated their overall satisfaction with acetaminophen compared with NSAIDs when both effectiveness and side effects were considered. RESULTS: Two-thirds of study participants had taken acetaminophen. About 37% of patients who had taken acetaminophen found it to be moderately or very effective and about 63% indicated that it was not effective or was only slightly effective. One-fourth of the patients found acetaminophen and NSAIDs to be equally effective, but >60% found acetaminophen to be much less effective or somewhat less effective. About 12% preferred acetaminophen to NSAIDs. When both effectiveness and side effects were considered together, 25% of the patients had no preference, 60% preferred NSAIDs, and 14% preferred acetaminophen. CONCLUSION: There was a considerable and statistically significant preference for NSAIDs compared with acetaminophen among the 3 groups of rheumatic disease patients. Although this preference decreased slightly with age and was less pronounced in OA patients, the preference was noted among all categories of patients and was not altered by disease severity. If safety and cost are not issues, there would hardly ever be a reason to recommend acetaminophen over NSAIDs, since patients generally preferred NSAIDs and fewer than 14% preferred acetaminophen. If safety and costs are issues, then the recommendation of the American College Rheumatology that acetaminophen be tried first seems correct, since 38.2% found acetaminophen to be as effective or more effective than NSAIDs. PMID- 10693879 TI - Scores for functional disability in patients with rheumatoid arthritis are correlated at higher levels with pain scores than with radiographic scores. AB - OBJECTIVE: To analyze correlations of functional disability scores with other measures of clinical status, in particular, Larsen radiographic scores and pain scores, in patients with rheumatoid arthritis (RA). METHODS: The functional capacity of 141 patients with RA (102 women, 39 men; median age 57 years; median disease duration 11.8 years; 83% rheumatoid factor positive) was assessed according to the Stanford Health Assessment Questionnaire (HAQ). Other variables studied included Larsen scores for radiographic damage of the small joints of the hands, wrists, and feet, pain scores by visual analog scale (VAS), Disease Activity Scores, general health scores by VAS, and Beck Depression Inventory (BDI) scores. RESULTS: The Spearman correlation coefficient comparing HAQ and Larsen scores was 0.277 (P = 0.001) and between HAQ and pain scores 0.652 (P < 0.001). In regression analysis, pain scores explained 41.4% of the variation in HAQ scores, normalized Larsen scores explained 7.3%, and BDI scores explained 5.5%; other variables were not significant in the model. CONCLUSION: Functional capacity scores of patients with RA are correlated at higher levels with pain scores than with radiographic scores of small joints. PMID- 10693880 TI - Recipients of hip replacement for arthritis are less likely to be Hispanic, independent of access to health care and socioeconomic status. AB - OBJECTIVE: To compare the proportion of Hispanics among recipients of hip replacements for primary articular disorders, recipients of knee replacements for the same reason, and persons hospitalized for other reasons. METHODS: Twelve of the 17 accredited hospitals in Bexar County, Texas, in which hip or knee replacement surgery is performed permitted us to review their medical records. From 1993 through 1995, 3,100 elective, non-fracture-related, hip or knee replacements were performed. These individuals were matched by age, sex, hospital, and month of admission with 4,604 persons hospitalized for other reasons. Age, sex, ethnic background, type of medical insurance, median household income by zip code of residence, joint replaced, and surgical diagnosis were abstracted from the medical records. The validity of variables abstracted from the medical records was tested by comparison with self-report data in 115 patients interviewed prior to elective hip or knee replacement surgery. RESULTS: During the study period, 2,275 subjects had a total knee replacement and 825 had a total hip replacement. Recipients of hip replacements were significantly less likely to be Hispanic than were recipients of knee replacements (19.5% versus 29.9%; odds ratio [OR] 0.57, 95% confidence interval [95% CI] 0.46-0.71; P < or = 0.001) or persons hospitalized for other reasons (29.4% Hispanic; OR 0.67, 95% CI 0.55-0.81). The under-representation of Hispanics was more pronounced among persons undergoing hip replacement for osteoarthritis compared with recipients of knee replacements for the same disease (OR 0.48, 95% CI 0.37-0.62). This pattern persisted after adjusting for age, sex, type of medical insurance, and median household income by the zip code of residence. Concordance between medical records and self-report data on ethnic background was high (kappa = 0.93). CONCLUSION: Recipients of hip replacement are less likely to be Hispanic than are other hospitalized persons with a similar level of access to care. The reasons for this under-representation probably involve factors in addition to lack of access to health care and low socioeconomic status. Further research is needed to understand the nature of such factors. PMID- 10693881 TI - Association of mild acetabular dysplasia with an increased risk of incident hip osteoarthritis in elderly white women: the study of osteoporotic fractures. AB - OBJECTIVE: To determine if acetabular dysplasia increases the risk of incident hip osteoarthritis (OA) among elderly white women. METHODS: Baseline and followup anteroposterior pelvic radiographs were obtained a mean of 8 years apart, and read for individual radiographic features (IRFs) of hip OA; summary grades (0-4) were then assigned based on the IRFs present. Acetabular dysplasia was defined by the results of measurements of the acetabular depth (<9 mm) or the center-edge angle (<30 degrees). Logistic regression analyses were performed to determine the association between acetabular dysplasia and incident hip OA, and all analyses were adjusted for age, current weight, body mass index, affected side, and investigational site. RESULTS: The odds ratios for the association of abnormal center-edge angle and acetabular dysplasia with incident hip OA were 3.3 (95% confidence interval 1.1-10.1) and 2.8 (95% confidence interval 1.0-7.9), respectively. CONCLUSION: Acetabular dysplasia, defined by a decrease in the center-edge angle, is associated with a modestly increased risk of incident hip OA in elderly white women. PMID- 10693882 TI - Drug-associated antineutrophil cytoplasmic antibody-positive vasculitis: prevalence among patients with high titers of antimyeloperoxidase antibodies. AB - OBJECTIVE: The triggers that induce antineutrophil cytoplasmic antibody (ANCA) positive vasculitis (APV) are largely unknown. However, there have been reports suggesting that hydralazine, propylthiouracil, and several other drugs may cause some cases of APV, and the majority of these cases have been associated with antimyeloperoxidase (anti-MPO) ANCA. Our experience led us to hypothesize that cases of high titers of anti-MPO antibodies are often drug-associated. METHODS: In this study, we determined the prevalence of exposure to hydralazine, propylthiouracil, and other drugs previously implicated in APV among 30 patients with vasculitis and the highest titers of anti-MPO antibodies newly detected in our laboratory between 1994 and 1998. The clinical, histologic, and other serologic features of these 30 patients were also examined. RESULTS: The 30 study patients accounted for 12% of the 250 new patients with APV and anti-MPO who were tested during the study period. All 30 study subjects had anti-MPO titers that were more than 12 times the median titer of the 250 patients. Ten (33%) of the 30 patients had been exposed to hydralazine and 3 (10%) had been exposed to propylthiouracil. An additional 5 patients (17%) had been exposed to 1 of the other previously reported candidate drugs: 2 to penicillamine, 2 to allopurinol, and 1 to sulfasalazine. One of the patients exposed to hydralazine had also been exposed to allopurinol. In all cases, the clinical and histologic findings were typical of APV. There was a strong association between the presence of antielastase and/ or antilactoferrin antibodies and exposure to candidate drugs. CONCLUSION: These data suggest that a sizable proportion of cases of APV with high titers of anti-MPO antibodies are drug-associated, especially following exposure to hydralazine or propylthiouracil. We recommend that the use of these drugs should be sought in cases of anti-MPO-positive vasculitis, particularly among patients with high titers of these antibodies. PMID- 10693883 TI - Epidemiology of systemic vasculitis: a ten-year study in the United Kingdom. AB - OBJECTIVE: To describe the epidemiology of the primary systemic vasculitides (PSV; Wegener's granulomatosis, Churg-Strauss syndrome, microscopic polyangiitis, polyarteritis nodosa) in a well-defined population over a 10-year period. METHODS: An inception cohort of patients from the Norwich Health Authority (NHA) who were >15 years of age and had PSV first diagnosed between January 1, 1988 and December 31, 1997 was collected. Incidence rates were adjusted for age and sex to the 1992 population. The prevalence of PSV in this cohort was estimated on December 31, 1997. Patients were classified according to the American College of Rheumatology 1990 vasculitis criteria and the Chapel Hill Consensus definitions. RESULTS: Eighty-two NHA residents fulfilled the inclusion criteria. There were 47 men and 35 women, with a mean age of 62.9 years (median 65.0 years). The overall annual incidence of PSV among NHA residents was 19.8/million (95% confidence interval [95% CI] 15.8-24.6). The point prevalence on December 31, 1997 was 144.5/million (95% CI 110.4-185.3). PSV was more common in males (23.5/million; 95% CI 17.3-31.3) than females (16.4/million; 95% CI 11.4-22.8). The age- and sex specific incidence showed a clear increase with age, with an overall peak in the 65-74 year age group (60.1/million). CONCLUSION: In our study population, the annual incidence of PSV is slowly increasing with time and the incidence is greatest in the elderly. PMID- 10693884 TI - The lupus erythematosus cell phenomenon: comparative analysis of antichromatin antibody specificity in lupus erythematosus cell-positive and -negative sera. AB - OBJECTIVE: To compare and investigate antihistone and antichromatin antibody responses as well as clinical variables in patients with systemic lupus erythematosus (SLE) who were either positive (LEC+) or negative (LEC-) for the lupus erythematosus (LE) cell phenomenon. METHODS: The binding properties of LEC+ and LEC- SLE sera to chromatin-associated nuclear antigens (histones H1, H2A, H2B, H3, H4; complexes of H2A-H2B, [H2A-H2B]-DNA, H1-DNA; total and H1-stripped chromatin; native and denatured DNA) were investigated. In addition, sera from patients with drug-induced lupus (by procainamide, hydralazine, or quinidine), as well as from patients with rheumatoid arthritis and osteoarthritis, were assessed. Enzyme-linked immunosorbent assay was used to detect specific antibody binding. RESULTS: Mirroring the important role of histone H1 in the formation of LE cells, anti-histone H1 reactivity was 8-fold higher in LEC+ sera than in LEC- sera. In addition, reactivities to most of the other antigens tested, i.e., other histones and histone-DNA complexes as well as chromatin and DNA, were significantly higher in LEC+ sera than in LEC- sera. All but 1 serum sample from the patients with drug-induced lupus were negative for LE cell formation as well as for anti-histone H1 reactivity, but displayed high antibody reactivities to histone-DNA complexes, including chromatin. Sera from patients with rheumatoid arthritis and osteoarthritis did not show significant binding to these antigens. When comparing the clinical features of LEC+ and LEC- SLE patients, severe organ involvement, including nephritis and central nervous system involvement, was common in the LEC+ group, but rare in the LEC- group. CONCLUSION: A positive LE cell phenomenon not only correlated with the presence of high anti-histone H1 antibody levels in SLE, but also indicated serologically and clinically active disease with major organ involvement. PMID- 10693885 TI - Cross-reactivity of antiidiotypic antibodies with DNA in systemic lupus erythematosus. AB - OBJECTIVE: To assess the functional relationship between antibodies reactive with DNA and antibodies reactive with the idiotypes (idiopeptides) of anti-DNA antibodies that are associated with systemic lupus erythematosus (SLE) in mice. METHODS: Antiidiotypic antibodies that appeared spontaneously in lupus mice, and others that were induced by immunization of normal, non-lupus mice, were analyzed for their reactivity by a range of direct binding, competition enzyme-linked immunosorbent assay (ELISA), and surface plasmon resonance (SPR) methods. Their reactions were assessed against synthetic peptides representing sequences of the V(H) region of anti-DNA monoclonal antibody (mAb) V-88, against the native mAb itself, and against mammalian DNA. RESULTS: In lupus mice, only sera with the highest reactivity against double-stranded DNA (dsDNA) also reacted with idiopeptides in ELISA, and this showed a strong statistical correlation. However, there was no significant relationship between antiidiotypic antibodies and anti single-stranded DNA antibodies. Immunization of (BALB/c x NZW)F1 mice with idiopeptides p64 (V(H) residues 64-80) or p92 (V(H) residues 92-105) induced antibodies that reacted not only against the respective peptides, but also against the native parent anti-DNA mAb V-88. Furthermore, the immune antiidiopeptide antibodies cross-reacted with dsDNA. Competition SPR experiments with the BIAcore system supported this observation. The binding reaction of V(H) peptide p64 (representing the CDR-H2/FR-H3 region of V-88) with antiidiopeptide antibodies was inhibited by dsDNA. CONCLUSION: This study identified a unique set of autoantibodies in SLE. They react with both autoantibody idiotopes and with dsDNA, thus having a dual specificity for 2 autoantigens. Because these antiidiotope antibodies arise naturally during the development of lupus disease, and because they bind also to dsDNA, this provides a mechanism whereby the production of anti-dsDNA antibodies is stimulated. These idiotopes on autoantibodies in lupus act as natural mimotopes for inducing anti-dsDNA antibodies, which, due to their dual specificity, may significantly contribute to the pathology of nephritis in SLE. PMID- 10693886 TI - Validation of the Sapporo criteria for antiphospholipid syndrome. AB - OBJECTIVE: To test the Sapporo criteria for the classification of the antiphospholipid syndrome (APS). METHODS: We classified 243 consecutive patients who had clinical diagnoses of primary APS (n = 49), secondary APS (n = 26), systemic lupus erythematosus (SLE) without clinical APS (n = 131), and lupus-like disease without clinical APS (n = 37). RESULTS: Sensitivity, specificity, positive predictive value, and negative predictive value were 0.71, 0.98, 0.95, and 0.88, respectively. False-negative findings were the result of patients being classified on the basis of minor criteria that were not included in the Sapporo criteria, such as livedo reticularis, thrombocytopenia, low-titer IgG or IgM anticardiolipin antibody, IgA anticardiolipin antibody, and anti-beta2 glycoprotein I antibody. Some patients with false-negative results were true seronegative cases. CONCLUSION: The Sapporo criteria for APS compare favorably with the American College of Rheumatology criteria for SLE and are usable for clinical studies. PMID- 10693887 TI - Systemic sclerosis sine scleroderma: demographic, clinical, and serologic features and survival in forty-eight patients. AB - OBJECTIVE: To describe the demographic, clinical, and laboratory features and natural history of patients with systemic sclerosis sine scleroderma (ssSSc), and to compare them with those of patients with SSc and limited cutaneous involvement (IcSSc). METHODS: The University of Pittsburgh Scleroderma Databank served as the data source. Patients were divided into those who had no skin thickening (ssSSc) and those who had skin thickening only distal to elbows or knees and/or of the face (IcSSc) during their disease course. These two groups were compared with regard to demographic characteristics, clinical, laboratory, and serologic features, and survival rates. Chi-square and Student's t-test analyses were performed, and Fisher's exact test was used as appropriate. RESULTS: Of 555 consecutive patients without diffuse cutaneous SSc, 48 (9%) had ssSSc and 507 (91%) had IcSSc. The ssSSc patients had a mean total disease duration of 18.6 years (15.1 years before study entry and 3.5 years of followup after study entry), and had not developed IcSSc or another connective tissue disease. Other than the absence of skin thickening, the ssSSc group had no significant differences in individual internal organ involvements, laboratory features, serum autoantibody type, or survival rate compared with patients with IcSSc. Within the category of lung involvement, patients with ssSSc had a significantly greater frequency of dyspnea with mild exertion or at rest, and a tendency toward reduced carbon monoxide diffusing capacity (<70% of predicted normal) and primary pulmonary arterial hypertension. Patients with IcSSc had significantly more frequent individual manifestations of digital pitting scars, digital-tip ulcers, telangiectasia, and calcinosis than those with ssSSc, in part related to increased time of observation. Puffy fingers and finger joint contractures were detected significantly more often in IcSSc patients. CONCLUSION: Systemic sclerosis sine scleroderma should be included in the spectrum of SSc with limited cutaneous involvement and should not be considered a distinct or separate disorder. PMID- 10693888 TI - Association of transforming growth factor beta1 genotype with spinal osteophytosis in Japanese women. AB - OBJECTIVE: To examine the possible relationship between a T-->C polymorphism at nucleotide position 29 of the transforming growth factor beta1 (TGFbeta1) gene and genetic susceptibility to radiographic spinal osteophytosis. METHODS: A total of 540 postmenopausal Japanese women were subjected to radiography of the spine and determination of bone mineral density (BMD) for the lumbar spine and total body. Changes in lumbar intervertebral discs were examined in 67 individuals with either osteoporosis or spinal osteophytosis by magnetic resonance imaging (MRI). TGFbeta1 genotype was determined with an allele-specific polymerase chain reaction assay. The serum concentration of TGFbeta1 was measured in 29 control subjects and in 36 patients with spinal osteophytosis. RESULTS: Among all study subjects, the prevalence of radiographic spinal osteophytosis in individuals with the CC genotype was greater than that in those with the TC or TT genotype. Logistic regression analysis, adjusted for age, height, body weight, time since menopause, smoking status, body fat, lean mass, and either lumbar spine or total body BMD, demonstrated that the frequency of the C allele in subjects with spinal osteophytosis was significantly greater than that in those without this condition. Comparison among control, osteoporosis, and spinal osteophytosis groups also revealed that the C allele was more prevalent in subjects with osteophytosis than in controls, even after adjustment for BMD. In contrast, as previously shown, the frequency of the C allele was lower in osteoporosis patients than in controls. The intervertebral disc area and the ratio of disc area to vertebral body area, as determined by MRI, were also lowest in subjects with the CC genotype. The serum concentration of TGFbeta1 increased with the number of C alleles in both controls and patients with spinal osteophytosis. CONCLUSION: The T29-->C polymorphism of the TGFbeta1 gene exhibited inverse patterns of association with genetic susceptibility to spinal osteophytosis and with osteoporosis. Although radiographic evaluation of osteophytes might not reflect the actual disease severity, the C allele, which protects against osteoporosis, may be a risk factor for genetic susceptibility to spinal osteophytosis. PMID- 10693889 TI - Rheumatoid arthritis exacerbation caused by exogenous interleukin-12. AB - Interleukin-12 (IL-12) is a pleiotropic cytokine with proinflammatory, immunoregulatory, antitumor, and antimetastatic properties. It plays a crucial role in the development of the Th1 response and subsequent interferon-gamma production and enhancement of cell-mediated cytotoxicity. Recently, IL-12 has been used as an experimental therapy for cancer. Given the multiple immunomodulatory properties of IL-12, there are potential concerns associated with its clinical use. Of special interest are the possible side effects of IL-12 therapy in patients with autoimmune diseases, especially those that are T cell mediated, such as rheumatoid arthritis (RA). We present a case of severe RA exacerbation caused by treatment with IL-12 for metastatic cervical cancer. This is the first reported case of RA flare caused by exogenous IL-12. PMID- 10693890 TI - The changing face of rheumatoid arthritis therapy: results of serial surveys. PMID- 10693891 TI - Diurnal rhythm of salivary cortisol levels in patients with recent-onset rheumatoid arthritis. PMID- 10693892 TI - Clinical images: Wegener's granulomatosis presenting as Pancoast tumor. PMID- 10693893 TI - Exacerbation of systemic lupus erythematosus after hepatitis B vaccination: comment on the article by Battafarano et al and the letter by Senecal et al. PMID- 10693894 TI - Absence of p53 mutation in Japanese patients with rheumatoid arthritis: comment on the article by Han et al. PMID- 10693895 TI - Severity of rheumatoid arthritis in Portuguese patients: comment on the article by Drosos et al and on the letter by Ronda et al. PMID- 10693896 TI - Clinical images: Leg pain and clubbing. PMID- 10693897 TI - Validation of the United States' version of the World Health Organization Quality of Life (WHOQOL) instrument. AB - In 1991, the World Health Organization initiated a project to simultaneously develop a quality of life (QOL) instrument in 15 countries: The World Health Organization Quality of Life (WHOQOL) instrument. This was intended as a generic QOL tool for use with patients across varying disease types, severities of illness, and cultural subgroups. The objective of the current study was to evaluate the WHOQOL-100 in the U.S., one of the original 15 participating countries. The WHOQOL is a 100-item self-report instrument consisting of 24 subscales within six domains: Physical, Psychological, Independence, Social, Environment, and Spiritual. Four additional items pertain to overall QOL/health. We tested the WHOQOL-100 (U.S. version) in a sample of 443 adults (n = 251 chronically ill, n = 128 healthy, and n = 64 childbearing) in the U.S. to test its reliability (internal consistency, test-retest), construct validity (convergent, discriminant), responsiveness, and factor structure. The WHOQOL-100 (U.S. version) has acceptable internal consistency (alpha range: 0.82-0.95 across domains) and reproducibility (ICC range: 0.83-0.96 at 2-week retest interval). It is responsive to change in clinical conditions, as evidenced by predicted score change (effect size) in women after childbirth. Construct validity was demonstrated by (1) its correlation with the Short Form-36 and Subjective Quality of Life Profile, and (2) its ability to discriminate between the diverse samples in this study. The conceptual structure was confirmed exactly with the exception of four facets that did not correlate most highly with the domains to which they were originally assigned, but these differences were minor. The WHOQOL measurement system is suitable for evaluating the QOL of adults in the U.S. The psychometric properties will be continually evaluated as more data become available in the U.S. PMID- 10693898 TI - Ready, set, stop: reflections on assessing quality of life and the WHOQOL-100 (U.S. version). World Health Organization Quality of Life. PMID- 10693899 TI - Quality of life measurement: will we ever be satisfied? PMID- 10693900 TI - Ready, set, stop, and standing. PMID- 10693901 TI - Shoulder disability questionnaire design and responsiveness of a functional status measure. AB - The Shoulder Disability Questionnaire (SDQ) is a measure covering 16 items designed to evaluate functional status limitation in patients with shoulder disorders. The responsiveness of the SDQ was evaluated for 180 patients with soft tissue shoulder disorders, without underlying systemic disorders. These patients participated in a randomized placebo-controlled trial, in which ultrasound and electrotherapy appeared to be ineffective as adjuvants to standardized exercise therapy. At baseline and at 6-week follow-up, patients completed the SDQ and rated severity of shoulder pain and their chief complaint, while a research physiotherapist rated severity of symptoms and restriction of mobility. At the 6 week follow-up, patients also rated overall change since baseline. According to the calibrated responsiveness ratio (CRR) and the area under the receiver operator characteristic curve (AUC) the SDQ discriminates accurately between self rated clinically stable and improved subjects. The presented results suggest that the SDQ is as responsive as the compared outcome measures, and therefore is ready for use in clinical trials. PMID- 10693902 TI - Comparing treatment valuations between and within subjects in clinical trials: does it make a difference? AB - Valuations may be sensitive to biases, especially if elicited alongside randomized clinical trials. We investigated the construction of valuations assigned by women who entered a randomized clinical trial and were allocated to in-hospital or domiciliary monitoring. Women assigned valuations (0-10 visual analogue scale) to the strategy they had been allocated to and to the alternative strategy. Valuations were expressed as a between-subject difference (assigned by the women allocated to the respective strategies) and as within-subject differences (assigned by all women). Domiciliary monitoring was valued higher by the women allocated to that strategy (P = 0.10). In-hospital monitoring was valued higher by the women allocated to that strategy (P = 0.02). The average within-subject differences differed by allocated strategy (P<0.01). The within subject valuation differences showed large variability between and within groups. An overrepresentation of women favoring domiciliary monitoring and asymmetric treatment experience inflated the average within-subject difference in the domiciliary group but deflated that difference in the in-hospital group. Neither the average between-subject difference nor the average within-subject differences are free of bias. Other study designs probably cannot prevent bias. Comparing within-subject and between-subject differences is instructive. PMID- 10693903 TI - Lessons learned from enrollment in the BEST study--a multicenter, randomized trial of group psychosocial support in metastatic breast cancer. AB - The BEST study, a multicenter randomized trial of group psychosocial support in metastatic breast cancer, had several unusual features that may have influenced recruitment, notably the group nature of the intervention and the need for close collaboration between medical and psychosocial investigators. The recruitment process was examined in light of these features. Establishment of study centers was facilitated by involvement of experienced medical investigators who had successfully collaborated in previous research projects. Systematic evaluation of potential subjects or direct recruitment by psychosocial investigators optimized recruitment; however, the group nature of the intervention prolonged recruitment. Overall, 652 women were approached and 237 (43.3% of those medically eligible) randomized. Using population-based estimates, 24.3% of women with metastatic breast cancer were assessed for the study and 8.7% randomized. A randomization ratio of 2:1 was required to form and maintain groups. Competing clinical trials were the greatest barrier to recruitment. Five lessons were learned during recruitment for this trial: (1) multicenter randomized trials of psychosocial interventions are feasible, even in very ill patients, (2) the use of a group intervention effectively increased the required sample size by 50%, (3) similarity of randomization rates suggests that generalizability of study results will probably be comparable to that of other randomized cancer trials, (4) multidisciplinary collaborations and involvement of experienced researchers facilitated enrollment, and (5) most challenges encountered in recruitment were similar to those seen in all clinical trials. PMID- 10693904 TI - Analytic strategies for recurrent events in epidemiologic studies: background and application to hospitalization risk in the elderly. AB - Due to the intraindividual dependence, specific analytic strategies are needed to assess risk factors for recurrent events. Although well established in the biostatistics literature, applications of these techniques are almost nonexistent in the field of epidemiology. The authors applied four different regression approaches for recurrent events (logistic, Poisson, and two different Cox proportional hazards regressions) to derive rate ratios of hospitalizations for various prognostic factors in a cohort of 2424 frail elderly. Over a median follow-up of 670 days, 3299 hospitalizations were observed in 1564 persons. Estimated rate ratios were similar in all four approaches and virtually identical in three. With all methods, confidence intervals of the rate ratios were considerably wider than with naive Poisson regression neglecting intraindividual dependence of events. Appropriate analysis of recurrent events is feasible with minor modifications of multivariable models familiar to epidemiologists and should no longer be neglected in epidemiologic research. In our setting, Poisson regression was the most convenient approach. PMID- 10693905 TI - Publications on diagnostic test evaluation in family medicine journals: an optimal search strategy. AB - Search strategies for articles reporting on diagnostic test evaluations have been subjected to less research than those in the domain of clinical trials. We set out to develop an optimal search strategy for publications on diagnostic test evaluations in general, that could be added to keywords describing the specific diagnostic test at issue. Nine Family Medicine journals were searched from 1992 through 1995 for primary publications on diagnostic test evaluation by hand searching and a Medline search strategy published earlier. Additionally, new search strategies have been developed with stepwise logistic regression, using Mesh terms and free text words related to diagnosis and test evaluation as independent variables. Hand searching identified 75 primary publications on diagnostic test evaluation from a total of 2467 primary publications. The previously published search strategy had a sensitivity of 73%, a specificity of 94%, and a positive predictive value of 29%. The most accurate new search strategy had a sensitivity of 80.0% (60/75; 95% CI: 71.0-89.1), a specificity of 97.3% (2327/2392; 95% CI; 96.6-97.9%), a positive predictive value of 48% (95% CI: 40-56) and diagnostic odds ratio of 149. All four new strategies used the Mesh term "sensitivity and specificity" (exploded with the Mesh terms "predictive value" and "ROC")and cumulatively added the text words "specificity," "false negative," "accuracy," and "screening." The search strategy using the Mesh term "sensitivity and specificity" (exploded) and the text words "specificity," "false negative," and "accuracy" has both higher sensitivity and specificity than the previously published strategy. The increase in specificity in three strategies reduces the absolute number of false-positive articles that have to be screened by 50-75%, compared to the number of false positives in the earlier strategy. PMID- 10693906 TI - A new surrogate variable for erectile dysfunction status in the Massachusetts male aging study. AB - Erectile dysfunction (ED) is the subject of a vast clinical literature, but little information has been gathered from random samples of the general public. The Massachusetts Male Aging Study (MMAS) addressed this important aspect of men's health. The MMAS was conducted in two waves, with baseline data collection in 1987-1989 and follow-up in 1995-1997. Subsequent to the baseline MMAS survey, a consensus developed that subjective measures are optimal for defining ED. Unfortunately, the baseline questionnaire did not ask subjects directly about their erectile functioning. Thus, we previously assigned the MMAS subjects a degree of impotence at baseline using a series of related questions, employing a discriminant formula constructed from a separate sample of urology clinic patients. At follow-up the men classified themselves directly in addition to answering the original series of related questions. In the present article, we report the results of a new discriminant function, based on the MMAS men at follow-up. We also compare the two methods and discuss our reasons for preferring the internally calibrated method. PMID- 10693907 TI - Within-center correlation in dialysis adequacy. AB - The purpose of this study was to determine whether patients with end stage renal disease treated with hemodialysis were correlated in dialysis adequacy within facilities. This was a retrospective analysis of dialysis adequacy based on urea reduction ratio (URR) values from 6969 patients dialyzed at 154 facilities. The within-center correlation was quantified using the between-center variation and the parameter p that was derived using ANOVA tables and mixed effects models. The variation in center means for URR was wider than expected for independent observations (52.9-76.1 versus 60.7-73.8, respectively). Furthermore, there was a significant within-center correlation in URR values across all facilities (p = 0.136, P<0.0001), which persisted after adjusting for patient specific covariates, facility characteristics, and state. In conclusion, there was a substantial within-center correlation in dialysis adequacy that reflected important center effects on the outcome of ESRD patients. PMID- 10693908 TI - Alcohol exposure and health services utilization in older veterans. AB - The objective of this study was to determine if increased alcohol exposure is associated with greater use of health services among older veterans. A total of 129 older veterans (> or =65 years old), receiving longitudinal care in a Veterans' Administration primary care clinic, were followed retrospectively for up to 42 months. Subjects were screened at baseline for problem drinking with the CAGE or the quantity-frequency questions from the Alcohol Use Disorders Identification Test (QF-AUDIT), and stratified by exposure into three categories: abstainers, social drinkers, and problem drinkers. Outcomes included total outpatient clinic visits, laboratory tests, radiologic and other technologic procedures, as well as acute care hospitalizations. For all subjects (N = 129), no association was found between alcohol exposure and use of any outpatient services. Among CAGE-screened (n = 62) abstainers, social drinkers, and problem drinkers, significant differences were found in the median number of laboratory tests (7.3 vs. 3.4 vs. 7.1, P = 0.004) and hospitalizations (0.3 vs. 0.0 vs. 0.3, P = 0.001) per patient year of follow-up. No exposure-outcome associations were present, however, among QF-AUDIT-screened subjects (n = 67). We were unable to demonstrate a consistent relationship between alcohol exposure and health services utilization. The effects of alcohol on older veterans' use of health services varied with the method used to measure alcohol exposure. Additional studies are needed to determine whether multiple, or possibly new, measures can more precisely define the health effects of alcohol in older populations. PMID- 10693909 TI - Serum C-reactive protein and fibrinogen concentrations and self-reported angina pectoris and myocardial infarction: findings from National Health and Nutrition Examination Survey III. AB - C-reactive protein may predict the risk of cardiovascular disease, but its association with angina pectoris in the general population has not been clearly established, however. We used data from National Health and Nutrition Examination Survey III conducted from 1988-1994 to examine the associations between serum C reactive protein and plasma fibrinogen concentrations and self-reported angina pectoris and myocardial infarction among 7,948 U.S. men and women aged 40 years and older. C-reactive protein and fibrinogen concentrations were moderately correlated (r = 0.43). After adjustment for age, sex, race or ethnicity, education, smoking status, systolic blood pressure, serum cholesterol, high density lipoprotein cholesterol, history of diabetes mellitus, body mass index, and physical activity, fibrinogen (but not C-reactive protein) concentration was significantly associated with self-reported angina pectoris. Neither fibrinogen or C-reactive protein concentrations were significantly associated with angina pectoris when entered in the model simultaneously. C-reactive protein and fibrinogen concentrations were positively associated with myocardial infarction when entered separately into models, but only C-reactive protein concentration was significantly associated with myocardial infarction when both variables were entered simultaneously. These cross-sectional data showed a significant positive association between C-reactive protein concentration and myocardial infarction but not self-reported angina pectoris in the U.S. population. PMID- 10693910 TI - Mortality, hospital discharges, and case fatality for pulmonary embolism in the Twin Cities: 1980-1995. AB - Pulmonary embolism (PE) causes substantial morbidity and mortality, but little information is available regarding recent secular trends for PE. This study determined trends for PE in adults ages 30 to 84 years in the Minneapolis-St. Paul metropolitan area from 1980 to 1995. The age-adjusted mortality rate for PE decreased approximately 50% during the study period; the rate ratio (RR) for 1992 95 compared to 1980-83 was 0.41 in men [95% confidence interval (CI) 0.31-0.55] and was 0.60 in women (95% CI 0.46-0.79). The hospital discharge rate for PE also decreased from 1980-83 to 1988-91 (RR 0.69, 95% CI 0.63-0.76 in men; RR 0.72, 95% CI 0.66-0.78 in women), but increased slightly between 1988-91 and 1992-95. The case fatality rate for PE decreased approximately 60% during the period (RR 0.38, 95% CI 0.28-0.51 in men; RR 0.37, 95% CI 0.28-0.50 in women). The PE trends were paralleled by declining hospital discharge rates for phlebitis and thrombophlebitis. These data support a changing natural history or possible improvements in the prevention, diagnosis, and treatment of PE between 1980 and 1995. PMID- 10693911 TI - Oxidative stress: introduction. PMID- 10693912 TI - What is oxidative stress? AB - Oxidative stress, defined as a disturbance in the balance between the production of reactive oxygen species (free radicals) and antioxidant defenses, is discussed in relation to its possible role in the production of tissue damage in diabetes mellitus. Important free radicals are described and biological sources of origin discussed, together with the major antioxidant defense mechanisms. Examples of the possible consequences of free radical damage are provided with special emphasis on lipid peroxidation. Finally, the question of whether oxidative stress is increased in diabetes mellitus is discussed. PMID- 10693913 TI - Negative consequences of glycation. AB - The Diabetes Control and Complications Trial (DCCT) established unequivocally that the effects of inadequate insulin action (as monitored by the level of hyperglycemia) are associated with the incidence and progression of diabetic retinopathy, nephropathy, and neuropathy. How does hyperglycemia induce the functional and morphologic changes that characterize diabetic complications? Increasing evidence points to a major role for sugar-derived advanced glycation end products (AGEs), which form inside and outside cells as a function of glucose concentration. Recent work in this area supports a central role for reactive oxygen species (ROS) in both the formation of AGEs, and in AGE-induced pathologic alterations in gene expression. Inhibition of ROS may also be centrally important in the action of drugs that prevent complications in diabetic animal models. PMID- 10693914 TI - Gliclazide scavenges hydroxyl and superoxide radicals: an electron spin resonance study. AB - The role of reactive oxygen species in diabetes and its complications are well known. Two therapeutic agents commonly used in the treatment of diabetes are the sulfonylureas gliclazide and glibenclamide. These drugs effectively reduce blood sugar in non-insulin-dependent diabetes mellitus, by augmenting insulin release. Gliclazide is known to be a general free radical scavenger as shown by its inhibition of o-dianisidine photo-oxidation. In this study, the effects of gliclazide and glibenclamide on free radicals were examined in vitro, using electron spin resonance spectroscopy. Superoxide radical (O2*-) generated from the hypoxanthine-xanthine oxidase system or hydroxyl radical (OH*) generated via the Fenton reaction were analyzed as spin adducts of 5,5-dimethyl-1-pyrroline-N oxide (DMPO). Gliclazide scavenged O2*- and OH* in a dose-dependent manner whereas glibenclamide was without effect. These findings suggest that gliclazide is not only effective in reducing blood sugar, but may also be beneficial as a result of inhibition of lipid and protein denaturation, which is believed to lead to the development of diabetic complications. PMID- 10693915 TI - Gliclazide decreases low-density lipoprotein oxidation and monocyte adhesion to the endothelium. AB - Increasing evidence implicates oxidized low-density lipoprotein (LDL) and advanced glycation end products (AGE) in the atherogenesis associated with diabetes mellitus. In the present study, we examined the in vitro effects of gliclazide on LDL oxidation and monocyte adhesion to endothelial cells induced by oxidized LDL and glycated albumin. To assess the clinical relevance of our in vitro findings, we also measured the effect on monocyte adhesion of gliclazide administration to type 2 diabetic patients. Incubation of human monocytes and endothelial cells with increasing concentrations of gliclazide (0 to 10 microg/mL) and native LDL (100 microg/mL) induced a dose-dependent diminution of cell-mediated LDL oxidation. Pretreatment of endothelial cells with gliclazide (0 to 10 microg/mL) before addition of native LDL (100 microg/mL) or glycated albumin (100 microg/mL) resulted in a dose-dependent diminution of oxidized LDL- and glycated albumin-induced monocyte adhesion to endothelial cells. In type 2 diabetic patients, administration of gliclazide inhibits the increased adhesiveness of monocytes to levels similar to those observed in control subjects. These results indicate that gliclazide is an antioxidant and suggest a beneficial effect of this drug in the prevention of atherosclerosis associated with type 2 diabetes. PMID- 10693916 TI - Free radical scavenging activity of sulfonylureas: a clinical assessment of the effect of gliclazide. AB - In long-term clinical studies the beneficial effects of gliclazide on platelets have been related to a reduction in oxidative stress. This property is because of gliclazide's free radical scavenging ability that relates to the unique amino azabicyclo-octare ring, which is grafted on to the sulfonylurea. During a blinded clinical trial, the possible effects of gliclazide were assessed in 30 non insulin-dependent diabetic patients. All patients had been treated for diabetes for more than 2 years (mean 8 years) and had been established on glibenclamide for over 2 years with or without adjunctive metformin therapy. Patients were studied for 6 months and randomized to continue either their present dose of glibenclamide or to be converted to an equipotent dose of gliclazide. Measurements were taken of hemostatic variables, the oxidative status of the plasma, and the redox status, both extracellularly as plasma albumin-thiols (PSH) and lipid peroxides, and intracellularly as red blood cell superoxide dismutase activity (SOD). At 3 months, diabetic control was unaltered, but there were significant improvements in the oxidative status of the gliclazide-treated patients. Lipid peroxides decreased (8.3 +/- 1.1 to 7.0 +/- .06 micromol/L, P < .01) and red blood cell SOD increased (135 +/- 21 to 152 +/- 36 microg/mL, P < .05). PSH levels were unaltered at 458 +/- 38 micromol/L, whereas they had decreased significantly in the glibenclamide patients (414 +/- 34 micromol/L, P < .05), resulting in a significant difference between the 2 treatment groups (P < .004). Platelet reactivity to collagen also improved in the gliclazide-treated patients, decreasing from 65.1% +/- 14% to 50.8% +/- 24% (P < .01). The reactivity of the platelets remained unaltered in the glibenclamide patients. At 6 months, the significant differences between the 2 treatment groups remained. Hence, gliclazide was shown in a clinical study to have free radical scavenging activity independent of glycemic control. PMID- 10693917 TI - Oxidative stress and glycemic regulation. AB - Oxidative stress is an acknowledged pathogenetic mechanism in diabetic complications. Hyperglycemia is a widely known cause of enhanced free radical concentration, whereas oxidative stress involvement in glycemic regulation is still debated. Glucose transport is a cascade of events starting from the interaction of insulin with its own receptor at the plasma membrane and ending with intracellular glucose metabolism. In this complex series of events, each step plays an important role and can be inhibited by a negative effect of oxidative stress. Several studies show that an acute increase in the blood glucose level may impair the physiological homeostasis of many systems in living organisms. The mechanisms through which acute hyperglycemia exerts these effects may be identified in the production of free radicals. It has been suggested that insulin resistance may be accompanied by intracellular production of free radicals. In adipocytes cultured in vitro, insulin increases the production of hydrogen peroxide, which has been shown to mimic the action of insulin. These data allow us to hypothesize that a vicious circle between hyperinsulinemia and free radicals could be operating: insulin resistance might cause elevated plasma free radical concentrations, which, in turn, might be responsible for a deterioration of insulin action, with hyperglycemia being a contributory factor. Data supporting this hypothesis are available. Vitamin E improves insulin action in healthy, elderly, and non-insulin-dependent diabetic subjects. Similar results can be obtained by vitamin C administration. PMID- 10693918 TI - Clinical relevance of the oxidative stress concept. AB - Subjects with type 2 diabetes have markedly increased rates of coronary heart disease (CHD) that are only partly explained by the increased levels of conventional cardiovascular risk factors such as total cholesterol, hypertension, and smoking. Although an increasing number of studies have suggested a role for glycemia in cardiovascular disease, considerable controversy remains. This issue may be resolved when the results of the UK Prospective Diabetes Study (UKPDS) are presented. One possible promising relatively new risk factor that may explain high levels of CHD in diabetic subjects is increased oxidative stress. Type 2 diabetic subjects have an increased preponderance of small dense low-density lipoprotein (LDL), which predisposes to the oxidation of LDL. Almost all studies show that diabetic subjects have increased oxidative stress. In addition, they may have lower levels of alpha-tocopherol. In most studies, increased oxidative stress has been associated with cardiovascular disease, although prospective data are lacking. If levels of oxidative stress are increased, what are the best levels to reduce it to? Improved glycemic control has been associated with decreased oxidative stress. Antioxidant replacement such as alpha-tocopherol may also be beneficial. Interestingly, some special properties of hypoglycemic agents have been described. Gliclazide has been reported to favorably affect both free radicals and platelet reactivity. Gliclazide may have a more favorable effect on tissue plasminogen activator (tPA) than tolbutamide. In conclusion, increased levels of oxidative stress may underlie some of the increased risk of cardiovascular disease in diabetic subjects. Interventions to decrease levels of oxidative stress by methods such as improved glycemic control, antioxidant therapy (ie, alpha-tocopherol), and gliclazide are indicated. PMID- 10693919 TI - Cytokines that signal through the leukemia inhibitory factor receptor-beta complex in the nervous system. AB - Cytokines that signal through the leukemia inhibitory factor (LIF) receptor, such as LIF and ciliary neuronotrophic factor, have a wide range of roles within both the developing and mature nervous system. They play a vital role in the differentiation of neural precursor cells into astrocytes and can prevent or promote neuronal differentiation. One of the conundrums regarding signalling through the LIF receptor is how it can have multiple, often conflicting roles in different cell types, such as enhancing the differentiation of astrocytes while inhibiting the differentiation of some neuronal cells. Factors that can modulate signal transduction downstream of cytokine signalling, such as "suppressor of cytokine signalling" proteins, which inhibit the JAK/STAT but not the mitogen activated protein kinase pathway, may therefore play an important role in determining how a given cell will respond to cytokine signalling. This review discusses the general effects of cytokine signalling within the nervous system. Special emphasis is placed on differentiation of neural precursor cells and the role that regulation of cytokine signalling may play in how a given precursor cell responds to cytokine stimulation. PMID- 10693920 TI - Amtyr1: characterization of a gene from honeybee (Apis mellifera) brain encoding a functional tyramine receptor. AB - Biogenic amine receptors are involved in the regulation and modulation of various physiological and behavioral processes in both vertebrates and invertebrates. We have cloned a member of this gene family from the CNS of the honeybee, Apis mellifera. The deduced amino acid sequence is homologous to tyramine receptors cloned from Locusta migratoria and Drosophila melanogaster as well as to an octopamine receptor cloned from Heliothis virescens. Functional properties of the honeybee receptor were studied in stably transfected human embryonic kidney 293 cells. Tyramine reduced forskolin-induced cyclic AMP production in a dose dependent manner with an EC50 of approximately 130 nM. A similar effect of tyramine was observed in membrane homogenates of honeybee brains. Octopamine also reduced cyclic AMP production in the transfected cell line but was both less potent (EC50 of approximately 3 microM) and less efficacious than tyramine. Receptor-encoding mRNA has a wide-spread distribution in the brain and subesophageal ganglion of the honeybee, suggesting that this tyramine receptor is involved in sensory signal processing as well as in higher-order brain functions. PMID- 10693921 TI - Differential effects of corticostriatal and thalamostriatal deafferentation on expression of the glutamate transporter GLT1 in the rat striatum. AB - This study compared the effects of the disruption of the two main presumably glutamatergic striatal inputs, the corticostriatal and thalamostriatal pathways, on GLT1 expression in the rat striatum, using in situ hybridization and immunohistochemistry. Unilateral ibotenate-induced thalamic lesion produced no significant changes in striatal GLT1 mRNA labeling and immunostaining as assessed at 5 and 12 days postlesion. In contrast, significant increases in both parameters were measured after bilateral cortical lesion by superficial thermocoagulation. GLT1 mRNA levels increased predominantly in the dorsolateral part of the striatum; there, the increases were significant at 5 (+84%), 12 (+101%), and 21 (+45%) but not at 35 days postlesion. GLT1 immunostaining increased significantly and homogeneously by 17-26% at 12 and 21 days postlesion. The increase in GLT1 expression at 12 days postlesion was further confirmed by western blot analysis; in contrast, a 36% decrease in glutamate uptake activity was measured at the same time point. These data indicate that striatal GLT1 expression depends on corticostriatal but not thalamostriatal innervation. Comparison of our results with previous data showing that cortical lesion by aspiration downregulates striatal GLT1 expression further suggests that differential changes in GLT1 expression, and thus presumably in glial cell function, may occur in the target striatum depending on the way the cortical neurons degenerate. PMID- 10693922 TI - Cyclic AMP-mediated regulation of GABA(A) receptor subunit expression in mature rat cerebellar granule cells: evidence for transcriptional and translational control. AB - Exposure of rat cerebellar granule cells cultured to maturity in vitro to forskolin, N6,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate (Bt2cAMP), and 3 isobutyl-1-methylxanthine (IBMX) down-regulated GABA(A) receptor alpha6 and beta3 subunits but up-regulated alpha1 and beta2 subunits with respect to vehicle treated controls. Dideoxyforskolin had no effect on subunit expression. Protein kinase A inhibitors, H-89 and Rp-adenosine 3',5'cyclic monophosphothioate, prevented these effects on alpha1 but not alpha6 subunit expression. Flunitrazepam-sensitive [3H]Ro 15-4513 binding sites were increased by 144 +/- 20% following forskolin treatment. [3H]Ro 15-4513 photoaffinity labelling showed that the GABA(A) receptor alpha1 subunit was the principal locus of the increased flunitrazepam-sensitive [3H]Ro 15-4513 binding. Forskolin decreased flunitrazepam insensitive [3H]Ro 15-4513 binding sites by 25 +/- 8% and resulted in a 20% decrease in the irreversible incorporation of radioactivity in the alpha6 subunit. Steady-state levels of GABA(A) receptor subunit mRNAs were determined by semiquantitative RT-PCR in forskolin-treated cultures. Forskolin, Bt2cAMP, and IBMX down-regulated GABA(A) receptor alpha6 subunit mRNA expression; alpha1 and beta3 mRNA levels were unaffected, whereas beta2 subunit mRNA was up-regulated. Dideoxyforskolin had no significant effect on alpha1, alpha6, beta2, and beta3 mRNA levels. Thus, in mature cerebellar granule cells, GABA(A) receptor expression can be regulated by intracellular cyclic AMP levels. This occurs at the level of gene transcription and/or translation by mechanisms that are only partially governed by protein kinase A. PMID- 10693923 TI - Phorbol ester activation of the neuronal nicotinic acetylcholine receptor alpha7 subunit gene: involvement of transcription factor Egr-1. AB - alpha-Bungarotoxin-sensitive neuronal nicotinic acetylcholine receptors from bovine adrenomedullary chromaffin cells are up-regulated by long-term exposure to phorbol esters. The rise in receptor density is paralleled by an increase in transcripts corresponding to the alpha7 subunit, which is a component of this receptor subtype. Transcriptional activation of the alpha7 subunit gene is evidenced in reporter gene transfection experiments, in which phorbol esters increase alpha7 promoter activity by up to 14-fold. About 80% of this activation is abolished when at least two of the three sites for the immediate-early transcription factor Egr-1, present in the proximal promoter region of the alpha7 subunit gene, are mutated simultaneously. In addition, phorbol esters elevate both Egr-1 mRNA and Egr-1 protein levels in chromaffin cells, whereas electrophoretic mobility shift assays show that the Egr-1 component of the complexes that originate at the alpha7 promoter increases in cells treated with phorbol esters. These results suggest that the transcription factor Egr-1 is involved in triggering expression of alpha-bungarotoxin-sensitive nicotinic receptors in response to external stimuli, such as the ones resulting from phorbol ester treatment, and support our previous hypothesis that the alpha7 subunit gene is one of the specific targets for Egr-1. PMID- 10693924 TI - A proteolipid protein-specific pre-mRNA (Ppm-1) contains intron 3 and is up regulated during myelination in the CNS. AB - Alternative splicing of the precursor for messenger RNA (pre-mRNA) is a common process utilised by higher eukaryotes to modulate gene expression. A single primary transcript may generate several proteins with distinct functions, expressed in tissue-specific, developmental patterns. This article describes an oligodendrocyte-specific pre-mRNA product of proteolipid protein gene (P/p) transcription, which is the precursor for P/p but not Dm20 mRNA in the CNS. This P/p-specific pre-mRNA (Ppm-1) includes the intact intron 3 of the P/p gene. It is first expressed during active myelination, and it localises to the nucleus of oligodendrocytes, in both normal and jimpy (jp) murine CNS. In addition to mouse, Ppm-1 is found also in rat and dog, but not toad or trout. Our work suggests that alternative splicing of the P/p gene primary transcript follows a branching pattern, resulting in the presence of at least one P/p isoform-specific pre-mRNA molecule, Ppm-1. Therefore, Dm20 mRNA may be the product of a divergent set of pre-mRNA splicing events. PMID- 10693925 TI - Major phosphorylation site (Ser55) of neurofilament L by cyclic AMP-dependent protein kinase in rat primary neuronal culture. AB - Ser55 of neurofilament L (NF-L) is reported to be partly phosphorylated in neurons and to be phosphorylated by cyclic AMP-dependent protein kinase (PKA). Bovine NF-L was phosphorylated by PKA in a low concentration of MgCl2 (0.3 mM) and digested by trypsin. Trypsin-digested fragments were assigned by MALDI/ TOF (matrix-assisted laser desorption and ionization/ time-of-flight) mass spectrometry. Phosphorylation sites were found at Ser41, Ser55, and Ser62 in the head region, with Ser55 considered the preferred site. A site-specific phosphorylation-dependent antibody against Ser55 rendered NF-L phosphorylated at Ser55 detectable in primary cultured rat neurons. One-hour treatment with 20 nM okadaic acid increased the phosphorylation level of Ser55, and co-treatment with 10 microM forskolin enhanced it. However, forskolin alone did not elevate the phosphorylation level. As a consequence, NF-L may be phosphorylated at Ser55 by PKA or by a PKA-like kinase in vivo; however, the phosphorylation level of Ser55 may be modulated by certain phosphatases sensitive to okadaic acid. PMID- 10693926 TI - Opioids transiently prevent activation of apoptotic mechanisms following short periods of serum withdrawal. AB - Opioids exert a proapoptotic effect on several normal and tumoral cells. The aim of the present article was to examine the effect of opioids on the PC12 rat pheochromocytoma cell line, a model for the study of chromaffin cell apoptosis. These cells produce delta- and kappa-opioid agonists and their receptors. Our results were as follows: The kappa- and delta2-opioid receptor agonists had a rapid but transient effect on apoptosis at 3 h, whereas mu opioids did not. The effect of opioids was reversible by the opioid antagonists naloxone and nor binaltorphimine. The effect of opioids was protective, suppressing serum deprivation-induced apoptosis to approximately 50% of controls. The protective effect of opioids on PC12 apoptosis was measurable only under serum deprivation. The effect of opioids was remarkably reproducible and highly constant in timing, which did not appear to depend on the duration of the preceding serum deprivation. Finally, opioids prevented the elevation of the Bcl-2 and Bak proteins following serum deprivation to the levels attained by serum supplementation. Our combined data suggest that opioids protect PC12 cells from entering a state of induced apoptosis following serum deprivation. PMID- 10693927 TI - Enhancement of dopaminergic neurotoxicity by the mercapturate of dopamine: relevance to Parkinson's disease. AB - The mechanisms that underlie dopaminergic neurodegeneration in Parkinson's disease (PD) are not known but have been proposed to involve oxidation of dopamine and related catechols. In other organ systems, cytotoxicity from catechol oxidation is profoundly influenced by mercapturate metabolism. Here we have tested the hypothesis that catechol thioethers produced in the mercapturic acid pathway may act as dopaminergic neurotoxins. A rat mesencephalic/neuroblastoma hybrid (MES) cell line was exposed to dopamine, 3,4 dihydroxyphenylacetic acid (DOPAC), or eight different catechol thioethers for up to 24 h, and the extent of apoptosis was quantified by a microculture kinetic assay. Apoptosis also was confirmed morphologically with Giemsa-stained cultures and by demonstration of internucleosomal DNA fragmentation. The results showed that dopamine at 5-50 microM produced concentration-dependent increases in the percentage of apoptotic MES cells. At 25 and 50 microM dopamine, the maximal proportions of apoptotic cells were detected at approximately 19 (20.7 +/- 2.0%) and 14 h (30.3 +/- 3.5%), respectively. None of the catechol thioethers (up to 5 microM) alone induced significant apoptosis in MES cells. However, when MES cells were incubated with dopamine (25 microM) and catechol thioethers (5 microM) to mimic pathological conditions, 5-S-N-acetylcysteinyldopamine, 5-S homocysteinyldopamine, and 5-S-homocysteinyl-DOPAC significantly increased the percentage of apoptotic cells compared with dopamine alone. These results suggest that mercapturate metabolism of endogenous catechols may yield products that facilitate dopaminergic neurodegeneration. PMID- 10693928 TI - ErbB-4 activation promotes neurite outgrowth in PC12 cells. AB - Neu differentiation factor (NDF; also known as neuregulin) induces a pleiotropic cellular response that is cell type-dependent. NDF and its receptor ErbB-4 are highly expressed in neurons, implying important roles in neuronal cell functions. In the present study we demonstrate that ErbB-4 receptors expressed in PC12 cells mediate NDF-induced signals and neurite outgrowth that are indistinguishable from those mediated by the nerve growth factor-activated Trk receptors. In PC12-ErbB-4 cells but not in PC12 cells, NDF induced an initial weak mitogenic signal and subsequently neurite outgrowth. The NDF-induced differentiation in PC12-ErbB-4 cells was mimicked by the pan-ErbB ligand betacellulin but not by other epidermal growth factor-like ligands. Thus, NDF and betacellulin mediate similar activities through the ErbB-4 receptor. Indeed, only these ligands induced strong phosphorylation of the ErbB-4 receptors. Neurite outgrowth induced by NDF in PC12 ErbB-4 cells was accompanied by sustained activation of mitogen-activated protein kinase (MAPK) and induction of the neural differentiation marker GAP-43. Inhibition of the MAPK kinase MEK or of protein kinase C (PKC) blocked NDF induced differentiation, whereas elevation of cyclic AMP levels enhanced the response. Taken together, these results indicate that neurite outgrowth induced by ErbB-4 in PC12 cells requires MAPK and PKC signaling networks. PMID- 10693929 TI - Brief exposure to neurotrophins produces a calcium-dependent increase in choline acetyltransferase activity in cultured rat septal neurons. AB - We demonstrate that brief (30-min) exposure of cultured embryonic rat septal neurons to neurotrophins (NTs) increases choline acetyltransferase (ChAT) activity by 20-50% for all tested NTs (nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, and neurotrophin-4, each at 100 ng/ml). The increase in ChAT activity was first detected 12 h after NT exposure, persisted at least 48 h, and was not mediated by increased neuronal survival or action potential activity. Under some conditions, the response to brief NT exposure was as great as that produced by continuous exposure. NT stimulation of ChAT activity was inhibited by inhibitors of p75- and Trk kinase-mediated signaling, by removal of extracellular Ca2+ during the period of NT exposure, and by buffering intracellular Ca2+ with BAPTA. Application of nerve growth factor and brain derived neurotrophic factor transiently increased [Ca2+] within a subpopulation of neurons. NT stimulation of ChAT activity was not affected significantly by cyclic AMP agonists or antagonists. These findings suggest that brief exposure to NTs can have a long-lasting effect on cholinergic transmission, and that this effect requires Ca2+, but not cyclic AMP. PMID- 10693930 TI - Glucocorticoids exacerbate the deleterious effects of gp120 in hippocampal and cortical explants. AB - Glucocorticoids (GCs), the adrenal steroids secreted during stress, can compromise the ability of hippocampal neurons to survive numerous necrotic insults. We have previously observed that GCs worsen the deleterious effects of gp120, the glycoprotein of the acquired immune deficiency syndrome virus, which can indirectly damage neurons and which is thought to play a role in the neuropathological features of human immunodeficiency virus infection. Specifically, GCs augment gp120-induced calcium mobilization, ATP depletion, decline in mitochondrial potential, and neurotoxicity in fetal monolayer cultures from a number of brain regions. In the present report, we demonstrate a similar gp120/GC synergy in adult hippocampal and cortical explants. We generated explants from rats that were either adrenalectomized, adrenally intact, or intact and treated with corticosterone to produce levels seen in response to major stressors. Metabolic rates in explants were then indirectly assessed with silicon microphysiometry, and cytosolic calcium concentrations were assessed with fura-2 fluorescent microscopy. We observed that basal levels of GCs tonically augment the disruptive effects of gp120 on metabolism in the CA1 cell field of the hippocampus and in the cortex. Moreover, raising GC concentrations into the stress range exacerbated the ability of gp120 to mobilize cytosolic calcium in a number of hippocampal cell fields. Finally, we observed that the synthetic GC prednisone had similarly exacerbating effects on gp120. Thus, GCs can worsen the deleterious effects of gp120 in a system that is more physiologically relevant than the fetal monolayer culture and in a region-specific manner. PMID- 10693931 TI - Apolipoprotein E exhibits isoform-specific promotion of lipid efflux from astrocytes and neurons in culture. AB - Many studies have shown that apolipoprotein E (apoE) plays important roles in maintaining intracellular lipid homeostasis in nonneuronal cells. However, little is known about the extracellular transport of lipids in the CNS. In this study, we determined whether and to what degree lipid efflux from astrocytes and neurons depended on apoE. Our results showed that exogenously added apoE promoted the efflux of cholesterol and phosphatidylcholine from both astrocytes and neurons in culture, resulting in the generation of high-density lipoprotein-like particles. The order of potency of the apoE isoforms as lipid acceptors was apoE2 > apoE3 = apoE4 in astrocytes and apoE2 > apoE3 > apoE4 in neurons. Treatment with brefeldin A, monensin, and a protein kinase C inhibitor, H7, abolished the ability of apoE to promote cholesterol efflux from cultured astrocytes, without altering apoE-mediated phosphatidylcholine efflux. In contrast, the efflux of both cholesterol and phosphatidylcholine promoted by apoE was abolished following treatment with heparinase or lactoferrin, which block the interaction of apoE with heparan sulfate proteoglycans (HSPGs) or low-density lipoprotein receptor related protein (LRP), respectively. This study suggests that apoE promotes lipid efflux from astrocytes and neurons in an isoform-specific manner and that cell surface HSPGs and/or HSPG-LRP pathway may mediate this apoE-promoted lipid efflux. PMID- 10693932 TI - Amyloid beta and amylin fibrils induce increases in proinflammatory cytokine and chemokine production by THP-1 cells and murine microglia. AB - Activated microglia surrounding amyloid beta-containing senile plaques synthesize interleukin-1, an inflammatory cytokine that has been postulated to contribute to Alzheimer's disease pathology. Studies have demonstrated that amyloid beta treatment causes increased cytokine release in microglia and related cell cultures. The present work evaluates the specificity of this cellular response by comparing the effects of amyloid beta to that of amylin, another amyloidotic peptide. Both lipopolysaccharide-treated THP-1 monocytes and mouse microglia showed significant increases in mature interleukin-1beta release 48 h following amyloid beta or human amylin treatment, whereas nonfibrillar rat amylin had no effect on interleukin-1beta production by THP-1 cells. Lipopolysaccharide stimulated THP-1 cells treated with amyloid beta or amylin also showed increased release of the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-6, as well as the chemokines interleukin-8 and macrophage inflammatory protein-1alpha and -1beta. THP-1 cells incubated with fibrillar amyloid beta or amylin in the absence of lipopolysaccharide also showed significant increases of both interleukin-1beta and tumor necrosis factor-alpha mRNA. Furthermore, treatment of THP-1 cells with amyloid fibrils resulted in an elevated expression of the immediate-early genes c-fos and junB. These studies provide further evidence that fibrillar amyloid peptides can induce signal transduction pathways that initiate an inflammatory response that is likely to contribute to Alzheimer's disease pathology. PMID- 10693933 TI - Fatty acid-activated K+ channels in autonomic neurons: activation by an endogenous source of free fatty acids. AB - Application of arachidonic acid evoked robust activation of large-conductance K+ channels in cell-attached and excised inside-out patches from acutely isolated chick ciliary ganglion neurons. A similar effect was produced by 5,8,11,14 eicosatetraynoic acid, a nonmetabolizable analogue of arachidonic acid. The unitary conductance of fatty acid-activated channels was 35-40 pS at +20 mV with physiological gradients of K+ and 165 pS at +20 mV with an extracellular K+ concentration of 37.5 mM and an intracellular K+ concentration of 150 mM. Gating of these channels in cell-attached patches was potentiated by membrane stretch. Channel gating evoked by both lipids was concentration-dependent, with detectable activation apparent at 4 microM in the majority of patches and maximal activation occurring between 32 and 64 microM. Gating was relatively voltage-independent. Large-conductance K+ channels were also activated in inside-out patches by the monounsaturated fatty acid 11-cis-eicosenoic acid but not by the fully saturated fatty acid arachidic acid. Application of 100 microM H2O2, an agent that activates cytosolic phospholipase A2, also caused activation of large-conductance K+ channels in intact neurons. The stimulatory effects of H2O2 were blocked by pretreatment with 20 microM 4-bromophenacyl bromide, an irreversible inhibitor of phospholipase A2. Therefore, mobilization of endogenous fatty acids can cause activation of large-conductance K+ channels in autonomic neurons. PMID- 10693934 TI - Differential regulation of cyclin D1 and D3 expression in the control of astrocyte proliferation induced by endothelin-1. AB - We have previously shown that the mitogenic effect of endothelin-1 (ET-1) in primary astrocytes is dependent on activation of both extracellular signal regulated kinase (ERK)- and cytoskeleton (CSK)-dependent pathways. In this study, we evaluated the contribution of each of these pathways to the expression and activation of proteins mediating cell cycle progression. Our results suggest that ET-1-induced expression of cyclins D1 and D3 is dependent on the ERK- and CSK dependent pathways, respectively; moreover, a decrease in the levels of the cyclin-dependent kinase inhibitor (CKI) p27 was observed as a consequence of ERK activation. Expression of both cyclins D1 and D3 together with a decrease in the p27 levels are essential for retinoblastoma protein (pRB) phosphorylation and cyclin A expression. Furthermore, the molecular events responsible for cell-cell contact inhibition of astrocyte proliferation were found to be independent of the mitogenic pathways leading to D-type cyclin expression. Cell growth arrest in confluent astrocytes was found to be correlated with increased expression of CKI p21, resulting in inhibition of D-type cyclin-associated pRB phosphorylation and cyclin A expression. Taken together, these results indicate that cyclins D1 and D3, which constitute the key mediators of the proliferative response of primary astrocytes to ET-1, are regulated by distinct signaling pathways. PMID- 10693935 TI - Turnover and down-regulation of GABA(A) receptor alpha1, beta2S, and gamma1 subunit mRNAs by neurons in culture. AB - Benzodiazepines (BZDs), barbiturates, ethanol, and general anesthetics potentiate the action of gamma-aminobutyric acid (GABA) at the type A GABA receptor (GABA(A)R) and have profound effects on mood, arousal, and susceptibility to seizures. GABA(A)R number and subunit mRNA levels change in animal models of epilepsy and anxiety and following exposure to GABA(A)R agonists and positive modulators, but the mechanism of receptor down-regulation remains unknown. Persistent exposure (48 h) of brain neurons in primary culture to GABA results in a 30% decrease in the levels of mRNA encoding the alpha1, beta2S, and gamma1 GABA(A)R subunit isoforms, which form a receptor enhanced by nonselective BZDs. Down-regulation of alpha1 mRNA (t1/2 = 8 h) precedes down-regulation of receptor number (t1/2 = 25 h), suggesting that GABA-induced GABA(A)R down-regulation is a consequence of decreased mRNA levels. The apparent half-life of the alpha1 mRNA in the presence of alpha-amanitin (9 h) is consistent with the time course of alpha1 mRNA down-regulation. Moreover, the stability of the alpha1, beta2S, and gamma1 subunit mRNAs is not altered by chronic GABA exposure. The results demonstrate that GABA(A)R subunit mRNA down-regulation is not a consequence of accelerated mRNA degradation and argue that GABA-induced GABA(A)R down-regulation is due to inhibition of transcription. PMID- 10693936 TI - Multiple calcium pathways induce the expression of SNAP-25 protein in chromaffin cells. AB - Incubation of bovine adrenal chromaffin cells in high K+ (38 mM) during 24-48 h enhanced 2.5 to five times the expression of SNAP-25 protein and mRNA, respectively. This increase was reduced 86% by furnidipine (an L-type Ca2+ channel blocker) but was unaffected by either omega-conotoxin GVIA (an N-type Ca2+ channel blocker) or -agatoxin IVA (a P/Q-type Ca2+ channel blocker). Combined blockade of N and P/Q channels with omega-conotoxin MVIIC did, however, block by 76% the protein expression. The inhibitory effects of fumidipine were partially reversed when the external Ca2+ concentration was raised from 1.6 to 5 mM. These findings, together with the fact that nicotinic receptor activation or Ca2+ release from internal stores also enhanced SNAP-25 protein expression, suggest that an increment of cytosolic Ca2+ concentration ([Ca2+]), rather than its source or Ca2+ entry pathway, is the critical signal to induce the protein expression. The greater coupling between L-type Ca2+ channels and protein expression might be due to two facts: (a) L channels contributed 50% to the global [Ca2+]i rise induced by 38 mM K+ in indo-1-loaded chromaffin cells and (b) L channels undergo less inactivation than N or P/Q channels on sustained stimulation of these cells. PMID- 10693937 TI - Immunocytochemical localization of beta-methylcrotonyl-CoA carboxylase in astroglial cells and neurons in culture. AB - Astroglia-rich primary cultures and brain slices rapidly metabolize branched chain amino acids (BCAAs), in particular leucine, as energy substrates. To allocate the capacity to degrade leucine oxidatively in neural cells, we have purified beta-methylcrotonyl-CoA carboxylase (beta-MCC) from rat liver as one of the enzymes unique for the irreversible catabolic pathway of leucine. Polyclonal antibodies raised against beta-MCC specifically cross-reacted with both enzyme subunits in liver and brain homogenates. Immunocytochemical examination of astroglia-rich rat primary cultures demonstrated the presence of beta-MCC in astroglial cells, where the enzyme was found to be located in the mitochondria, the same organelle that the mitochondrial isoform of the BCA(A) aminotransferase (BCAT) is located in. This colocalization of the two enzymes supports the hypothesis that mitochondrial BCAT is the isoenzyme that in brain energy metabolism prepares the carbon skeleton of leucine for irreversible degradation in astrocytes. Analysis of neuron-rich primary cultures revealed also that the majority of neurons contained beta-MCC. The presence of beta-MCC in most neurons demonstrates their ability to degrade the alpha-ketoisocaproate that could be provided by neighboring astrocytes or could be generated locally from leucine by the action of the cytosolic isoform of BCAT that is known to occur in neurons. PMID- 10693938 TI - Immunoprecipitation of high-affinity, guanine nucleotide-sensitive, solubilized mu-opioid receptors from rat brain: coimmunoprecipitation of the G proteins G(alpha o), G(alpha i1), and G(alpha i3). AB - Antibodies directed against the C-terminal and the N-terminal regions of the mu opioid receptor were generated to identify the G proteins that coimmunoprecipitate with the mu receptor. Two fusion proteins were constructed: One contained the 50 C-terminal amino acids of the mu receptor, and the other contained 61 amino acids near the N terminus of the receptor. Antisera directed against both fusion proteins were capable of immunoprecipitating approximately 70% of solubilized rat brain mu receptors as determined by [3H][D-Ala2,N-Me Phe4,Gly-ol5]-enkephalin ([3H]DAMGO) saturation binding. The material immunoprecipitated with both of the antisera was recognized as a broad band with a molecular mass between 60 and 75 kDa when screened in a western blot. Guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) had an EC50 of 0.4 nM in diminishing [3H]DAMGO binding to the immunoprecipitated pellet. The ratio of G proteins to mu receptors in the immunoprecipitated material was 1:1. When the material immunoprecipitated with affinity-purified antibody was screened for the presence of G protein a subunits, it was determined that G(alpha)o, G(alpha)i1, G(alpha)i3, and to a lesser extent G(alpha)i2, but not G(alpha)s or G(alpha)q11, were coimmunoprecipitated with the mu receptor. Inclusion of GTPgammaS during the immunoprecipitation process abolished the coimmunoprecipitation of G proteins. PMID- 10693939 TI - Attenuation and recovery of evoked overflow of striatal serotonin in rats treated with neurotoxic doses of methamphetamine. AB - Repeated administration of methamphetamine to animals can lead to long-lasting decreases in striatal monoamine content. In the present study, the effects of neurotoxic doses of methamphetamine on basal and evoked overflow of striatal serotonin and of its primary metabolite 5-hydroxyindoleacetic acid were examined in awake rats using in vivo microdialysis. Male Fischer-344 rats were administered methamphetamine (5 mg/kg, s.c.) or saline four times in 1 day at 2-h intervals. Microdialysis studies were carried out 1 week, 1 month, and 6 months later. At 1 week posttreatment there were significant decreases in potassium- and amphetamine-evoked overflow of serotonin in the striatum of the methamphetamine treated animals. Basal extracellular levels of 5-hydroxyindoleacetic acid but not of serotonin were also decreased. Evoked overflow of serotonin recovered by 1 month, and extracellular levels of 5-hydroxyindoleacetic acid had recovered by 6 months. Tissue levels of serotonin and 5-hydroxyindoleacetic acid were decreased at 1 week posttreatment but back to control levels by 1 month after treatment. These results indicate that presynaptic serotonergic functioning is attenuated in the striatum of rats treated 1 week earlier with neurotoxic doses of methamphetamine. However, in the model used, the changes are transient, and recovery can occur within 1-6 months posttreatment. PMID- 10693940 TI - Quantitative and statistical analysis of the shape of amperometric spikes recorded from two populations of cells. AB - Previously used methods of comparing amperometric spike characteristics from two separate groups of cells have entailed pooling all the values for a spike characteristic from each group of cells and then statistically comparing the two samples. Although this approach has indicated that there are significant differences between the spike characteristics from coloboma and control mouse chromaffin cells, the results are not consistent between experiments. We have reexamined the assumptions of the statistical tests used as well as the variability inherent in amperometric data measured from two groups of cells. Our findings indicate that when comparing amperometric spike characteristics between groups of cells, it is more appropriate to compare samples of mean spike values. This method consistently indicates that there is no difference between coloboma and control amperometric spikes. These results have been validated by using samples of mean spike characteristics to detect changes in the shape of amperometric spikes from both mouse chromaffin cells at 37 degrees C and PC12 cells previously exposed to 50 microM L-3,4-dihydroxyphenylalanine and by the use of an additional analysis method, the nested ANOVA. Together, these results indicate that pooled samples of amperometric spike characteristics can give results that may confound the interpretation of amperometric data. PMID- 10693942 TI - Elevated N1-acetylspermidine levels in gerbil and rat brains after CNS injury. AB - The polyamine system is very sensitive to different pathological states of the brain and is perturbed after CNS injury. The main modifications are significant increases in ornithine decarboxylase activity and an increase in tissue putrescine levels. Previously we have shown that the specific polyamine oxidase (PAO) inhibitor N1,N4-bis(2,3-butadienyl)-1,4-butanediamine (MDL 72527) reduced the tissue putrescine levels, edema, and infarct volume after transient focal cerebral ischemia in spontaneously hypertensive rats and traumatic brain injury of Sprague-Dawley rats. In the present study, N1-acetyl-spermidine accumulation was greater in injured brain regions compared with sham or contralateral regions following inhibition of PAO by MDL 72527. This indicates spermidine/spermine-N1 acetyltransferase (SSAT) activation after CNS injury. The observed increase in N1 acetylspermidine levels at 1 day after CNS trauma paralleled the decrease in putrescine levels after treatment with MDL 72527. This suggests that the increased putrescine formation at 1 day after CNS injury is mediated by the SSAT/PAO pathway, consistent with increased SSAT mRNA after transient ischemia. PMID- 10693943 TI - Peptide fragments of beta-amyloid precursor protein: effects on parallel fiber Purkinje cell synaptic transmission in rat cerebellum. AB - The effects of peptide fragments of the beta-amyloid precursor protein (betaAPP) on parallel fiber (PF)-Purkinje cell synaptic transmission in the rat cerebellum were examined. Transient inward currents associated with calcium influx were induced by localized applications of the 105-amino acid carboxy-terminal fragment (CT105) of betaAPP to discrete dendritic regions of intact Purkinje cells. betaAPP and the amyloid beta (Abeta) peptide fragments Abeta1-16, Abeta25-35, and Abeta1-42 had little or no effect. Inward currents were also observed following applications of CT105 to isolated patches of somatic Purkinje cell membrane. All five proteins/peptides induced some depression of alpha-amino-3-hydroxy-5-methyl 4-isoxazole propionate (AMPA) receptor-mediated synaptic transmission between PFs and Purkinje cells, through a combination of pre- and postsynaptic effects. CT105 induced the greatest depression, which spread to distant synapses following local application and which was prevented by inhibition of nitric oxide synthase. These data indicate that CT fragments of the betaAPP can modulate AMPA-mediated glutamatergic synaptic transmission in the cerebellar cortex. These fragments may therefore be considered alternative candidates for some of the neurotoxic effects of Alzheimer's disease. PMID- 10693941 TI - Up-regulation of base excision repair activity for 8-hydroxy-2'-deoxyguanosine in the mouse brain after forebrain ischemia-reperfusion. AB - The repair enzyme 8-oxoguanine glycosylase/ apyrimidinic/apurinic lyase (OGG) removes 8-hydroxy-2'deoxyguanosine (oh8dG) in human cells. Our goal was to examine oh8dG-removing activity in the cell nuclei of male C57BL/6 mouse brains treated with either forebrain ischemia-reperfusion (FblR) or sham operations. We found that the OGG activity in nuclear extracts, under the condition in which other nucleases did not destroy the oligodeoxynucleotide duplex, excised oh8dG with the greatest efficiency on the oligodeoxynucleotide duplex containing oh8dG/dC and with less efficiency on the heteroduplex containing oh8dG/dT, oh8dG/dG, or oh8dG/dA. This specificity was the same as for the recombinant type 1 OGG (OGG1) of humans. We observed that the OGG1 peptide and its activity in the mouse brain were significantly increased after 90 min of ischemia and 20-30 min of reperfusion. The increase in the protein level and in the activity of brain OGG1 correlated positively with the elevation of FblR-induced DNA lesions in an indicator gene (the c-fos gene) of the brain. The data suggest a possibility that the OGG1 protein may excise oh8dG in the mouse brain and that the activity of OGG1 may have a functional role in reducing oxidative gene damage in the brain after FblR. PMID- 10693944 TI - Substrate-bound beta-amyloid peptides inhibit cell adhesion and neurite outgrowth in primary neuronal cultures. AB - Accumulation of the beta-amyloid protein (Abeta) in the brain is an important step in the pathogenesis of Alzheimer's disease. However, the mechanism of Abeta toxicity remains unclear. Abeta can bind to the extracellular matrix, a structure that regulates adhesive events such as neurite outgrowth and synaptogenesis. The binding of Abeta to the extracellular matrix suggests that Abeta may disrupt cell substrate interactions. Therefore, the effect of substrate-bound Abeta on the growth of isolated chick sympathetic and mouse cortical neurons was examined. Abeta1-40 and Abeta1-42 had dose-dependent effects on cell morphology. When tissue culture plates were coated with 0.1-10 ng/well Abeta, neurite outgrowth increased. Higher amounts of Abeta peptides (> or =3 microg/well) inhibited outgrowth. The inhibitory effect was related to aggregation of the peptide, as preincubation of Abeta1-40 for 24 h at 37 degrees C (a process known to increase amyloid fibril formation) was necessary for inhibition of neurite outgrowth. Abeta29-42, but not Abeta1-28, also inhibited neurite outgrowth at high concentrations, demonstrating that the inhibitory domain is located within the hydrophobic C-terminal region. Abeta1-40, Abeta1-42, and Abeta29-42 also inhibited cell-substrate adhesion, indicating that the effect on neurite outgrowth may have been due to inhibition of cell adhesion. The results suggest that accumulation of Abeta may disrupt cell-adhesion mechanisms in vivo. PMID- 10693945 TI - Increased generation of alternatively cleaved beta-amyloid peptides in cells expressing mutants of the amyloid precursor protein defective in endocytosis. AB - The subcellular location of the secretases processing the beta-amyloid precursor protein (APP) is not established yet. We analyzed the generation of the beta amyloid peptide (Abeta) in human embryonic kidney 293 cell lines stably expressing wild-type and noninternalizing mutants of human APP. APP lacking the entire cytoplasmic domain or with both tyrosine residues of the motif GYENPTY mutated to alanine showed at least fivefold reduced endocytosis. In these cell lines, the production of Abeta1-40 was substantially reduced, but accompanied by the appearance of two prominent alternative Abeta peptides differing at the amino termini. Based on antibody reactivity and mobility in high-resolution gels in comparison with defined Abeta fragments, these peptides were identified as Abeta3 40 and Abeta5-40. Notably, these alternative Abeta peptides were not generated when the APP mutants were retained in the early secretory pathway by treatment with brefeldin A. These results indicate that the alternative processing is the result of APP accumulation at the plasma membrane and provide evidence of distinct beta-secretase activities. Cleavage amino-terminal to position 1 of Abeta occurs predominantly in endosomes, whereas the processing at positions 3 or 5 takes place at the plasma membrane. PMID- 10693946 TI - Immediate early gene expression in PC12 cells exposed to lead: requirement for protein kinase C. AB - We previously demonstrated induction of c-fos mRNA in PC12 cells exposed to lead that was dependent on new transcription. In the current work, we examined two signal transduction mechanisms that are activated by lead and have been shown to mediate induction of c-fos mRNA. One mechanism involves protein kinase C, and the other requires calmodulin-dependent protein kinase II. Significant increases in the levels of c-fos, c-jun, and egr-1 but not NGFIB mRNA were observed in PC12 cells exposed to lead or phorbol 12-myristate 13-acetate. In contrast, PC12 cells depolarized with 56 mM K+ displayed an increase in c-fos, egr-1, and NGFIB but not c-jun mRNA. Similar to other activators of protein kinase C, lead increased AP-1 and Egr-1 DNA binding activity. Additionally, lead increased luciferase activity in cerebellar granule cells transfected with an AP-1 luciferase reporter construct. Lead did not increase c-fos mRNA in PC12 cells that were depleted of protein kinase C by a 24-h treatment with phorbol 12,13-dibutyrate or incubated with the protein kinase C inhibitor H-7. In contrast, an inhibitor of calmodulin dependent protein kinase, KN-62, and an inhibitor of calmodulin, W-7, did not block the induction of c-fos mRNA by lead. An increase in serum-response element DNA-binding activity was observed in nuclear extracts from PC12 cells exposed to lead. It is interesting that lead activated protein kinase C isoforms delta and epsilon, but not isoforms alpha and beta. In conclusion, lead appears to induce the expression of immediate early genes by a mechanism that requires protein kinase C. PMID- 10693947 TI - Nigrostriatal reduction of aromatic L-amino acid decarboxylase activity in MPTP treated squirrel monkeys: in vivo and in vitro investigations. AB - Aromatic L-amino acid decarboxylase (AAAD) activity was examined in vivo with positron emission tomography (PET) using 6-[18F]fluoro-L-DOPA (FDOPA) in squirrel monkeys lesioned with graded doses of the neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). In vitro biochemical determinations of AAAD activity in caudate, putamen, substantia nigra, and nucleus accumbens were performed in the same animals to establish a direct comparison of in vivo and in vitro measurements. In vivo and in vitro AAAD activities in caudate/ putamen were substantially reduced in animals treated with the highest dose of MPTP (2.0 mg/kg). The percent change in the striatal FDOPA uptake (K(i)) and decarboxylation rate constant (k3) values resulting from MPTP treatment showed highly significant correlations with in vitro-determined AAAD activities. However, decarboxylase rates within individual animals presented as approximately 10-fold difference between in vivo and in vitro values. Lower in vivo k3 measurements may be attributed to several possibilities, including transport restrictions limiting substrate availability to AAAD within the neuron. In addition, reductions in AAAD activity in the substantia nigra did not parallel reductions in AAAD activity within the striatum, supporting the notion of a nonlinear relationship between nigrostriatal cell degeneration and terminal losses. This work further explores the role of AAAD in Parkinson's disease, a more important factor than previously thought. PMID- 10693948 TI - Chronic mitochondrial inhibition induces selective motoneuron death in vitro: a new model for amyotrophic lateral sclerosis. AB - Evidence is increasing that mitochondrial dysfunction is involved in amyotrophic lateral sclerosis, a neurodegenerative disease characterized by selective motoneuron death. To study the role of mitochondrial dysfunction in the pathways leading to motoneuron death, we developed an in vitro model of chronic motoneuron toxicity, based on malonate-induced inhibition of complex II in the mitochondrial electron transport chain. Treatment with malonate resulted in a dose-dependent decrease in cellular ATP levels. We observed that motoneurons were significantly more vulnerable to mitochondrial inhibition than control neurons in the dorsal horn. We could reproduce this dose-dependent phenomenon with the complex IV inhibitor sodium azide. The free radical scavenger alpha-phenyl-N-tert butylnitrone, the AMPA/kainate receptor blocker 6-cyano-7-nitroquinoxaline-2,3 dione, and riluzole, a drug that is currently used for the treatment of amyotrophic lateral sclerosis, were protective against malonate-induced motoneuron death. Furthermore, the caspase inhibitors N-benzyloxycarbonyl-Val-Ala Asp-fluoromethyl ketone and z-Asp-Glu-Val-Asp-fluoromethyl ketone were both protective against malonate toxicity. Our model shows that chronic mitochondrial inhibition leads to selective motoneuron death, which is most likely apoptotic. PMID- 10693949 TI - GluR1 glutamate receptor subunit is regulated differentially in the primate basal ganglia following nigrostriatal dopamine denervation. AB - Nigrostriatal dopaminergic denervation is associated with complex changes in the functional and neurochemical anatomy of the basal ganglia. The excitatory neurotransmitter glutamate mediates neural signaling at crucial points of this circuitry, and glutamate receptors are differentially distributed in the basal ganglia. Available evidence suggests that the glutamatergic corticostriatal and subthalamofugal pathways become overactive after nigrostriatal dopamine depletion. In this study, we have analyzed the regulation of the GluR1 subunit of the a-amino-3-hydroxy-5-methyl-4-isoxazole propionate glutamate receptor in the basal ganglia of primates following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced dopamine denervation. The dopamine denervation resulted in distinct alterations in GluR1 distribution: (1) GluR1 protein expression was markedly increased in caudate and putamen, and this was most pronounced in the striosomes; (2) GluR1 protein was altered minimally in subthalamic nucleus; (3) expression of GluR1 was down-regulated in the globus pallidus by 63% and in the substantia nigra by 57%. The down-regulation of GluR1 expression in the output nuclei of the basal ganglia, the internal segment of the globus pallidus and the substantia nigra pars reticulata, may be a compensation for the overactive glutamatergic input from subthalamic nucleus, which arises after striatal dopamine denervation. Our results indicate that the glutamatergic system undergoes regulatory changes in response to altered basal ganglia activity in a primate model of Parkinson's disease. Targeted manipulation of the glutamatergic system may be a viable approach to the symptomatic treatment of Parkinson's disease. PMID- 10693950 TI - Neuroprotective mechanism of glial cell line-derived neurotrophic factor in mesencephalic neurons. AB - Glial cell line-derived neurotrophic factor (GDNF) provides neuroprotection, but its neuroprotective mechanism has not been resolved. We investigated the neuroprotective mechanism of GDNF using primary culture of the rat mesencephalon. Bleomycin sulfate (BLM) and L-buthionine-[S,R]-sulfoximine (BSO) caused apoptosis in both dopaminergic and nondopaminergic neurons, as revealed by the presence of chromatin condensation, and positive staining by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL). GDNF preincubation blocked the neurotoxicity and reduced the number of the TUNEL positive cells caused by BLM and BSO exposure. In contrast, GDNF did not provide neuroprotection against glutamate toxicity, which was not accompanied by these apoptotic features. The neuroprotection was mediated by phosphatidylinositol 3 kinase, an effector downstream from c-Ret, because it was blocked by LY294002. GDNF pretreatment caused up-regulation of Bcl-2 and Bcl-x. Furthermore, GDNF suppressed oxygen radical accumulation caused by BLM. Apoptosis induced by BLM and BSO was blocked by a caspase-3 inhibitor. Caspase-3 activity was elevated by BLM and suppressed by GDNF pretreatment. These findings indicate that GDNF has no effect on necrosis but exerts protection against apoptosis by activation of phosphatidylinositol 3-kinase and the subsequent up-regulation of Bcl-2 and Bcl x, which suppresses accumulation of oxygen radicals followed by caspase-3 activation. PMID- 10693951 TI - Reduction in TrkA-immunoreactive neurons is not associated with an overexpression of galaninergic fibers within the nucleus basalis in Down's syndrome. AB - Down's syndrome (DS) individuals develop neuropathological features similar to Alzheimer's disease (AD), including degeneration of cholinergic basal forebrain (CBF) neurons. In AD a reduction in CBF/trkA-containing neurons has been suggested to trigger a hyperexpression of galaninergic fibers within the nucleus basalis subfield of the basal forebrain. The present study examined the interrelationship between reductions in CBF/trkA-containing neurons and the overexpression of galaninergic fibers within the nucleus basalis in DS. Within the nucleus basalis stereologic evaluation revealed a 46% reduction in the number of trkA-immunopositive neurons, whereas optical density measurements displayed a nonsignificant 18% reduction in neuronal trkA immunoreactivity in DS as compared with age-matched controls. Western blot analysis also showed a significant reduction in cortical trkA protein levels in DS. A semiquantitative examination of galaninergic fibers in the nucleus basalis revealed only a modest hypertrophy of galaninergic fibers within the nucleus basalis in DS. The present findings indicate a significant reduction in trkA within the nucleus basalis and cortex with only a moderate hypertrophy of galaninergic fibers in DS. These observations suggest that DS may not be an exact genetic model for investigation of changes in the AD basal forebrain. PMID- 10693952 TI - C3-fullero-tris-methanodicarboxylic acid protects cerebellar granule cells from apoptosis. AB - Buckminsterfullerenols were recently investigated for their protective properties in different models of acute and chronic neurodegeneration. We tested C3-fullero tris-methanodicarboxylic acid in our in vitro model of apoptotic neuronal death, which consists of shifting the culture K+ concentration from 25 to 5 mM for rat cerebellar granule cells. The impairment of mitochondrial respiratory function as well as chromatin derangement and fragmentation of DNA in apoptotic oligonucleosomes that occur in these conditions were protected by this compound in a concentration-dependent way. To assess whether antioxidant activity could account for the rescue of cerebellar granule cells from apoptosis, we tested the fullerene derivative under FeSO4-induced oxidative stress and found significant protection. Thus, we visualized membrane and cytoplasmic peroxides and reactive oxygen species and found a significant reduction of the species after 24 h in 5 mM K+ with the fullerene derivative. Such evidence suggests that this compound exerts a protective role in cerebellar granule cell apoptosis, likely reducing the oxidative stress. PMID- 10693953 TI - Diminished glutathione levels cause spontaneous and mitochondria-mediated cell death in neurons from trisomy 16 mice: a model of Down's syndrome. AB - It has been suggested that the increased neuronal death in cultures from trisomy 16 (Ts16) mice, a model of Down's syndrome, might result from a diminished concentration of reduced glutathione (GSH). In this study we used microfluorometric techniques to investigate the effect of GSH levels on neuronal survival in diploid and Ts16 cultures. Addition of the GSH precursors cysteine and cystine and the antioxidant tocopherol to the culture medium increased the GSH concentration up to 126.0% in diploid and up to 111.9% in Ts16 neurons. Moreover, we observed a reduced spontaneous neuronal death rate in diploid and Ts16 cultures. Following the application of 50-100 microM glutamate to culture medium, we found a GSH increase in the presence of cysteine, cystine, tocopherol, and cyclosporin A, an inhibitor of mitochondrial permeability transition (diploid, 105.8-110.8%; Ts16, 83.1-96.3%). However, only tocopherol and cyclosporin A had a protective effect on glutamate-induced neuronal death. The results suggest that reduced GSH levels affect the increase of a spontaneous and a mitochondria-mediated, cyclosporin A-sensitive type of neuronal cell death. Therefore, elevating intracellular GSH concentration may have neuroprotective effects in Down's syndrome and Alzheimer's disease. PMID- 10693954 TI - Involvement of caspase-3-like protease in the mechanism of cell death following focally evoked limbic seizures. AB - The cysteine protease caspase-3 may be involved in the mechanism of cell death following seizures. Using a rat model of focally evoked limbic epilepsy with continuous electroencephalography monitoring, we investigated seizure-induced changes in caspase-3 protein expression and processing, enzyme activity, and the in vivo effect of caspase-3 inhibition. Seizures were induced by intraamygdaloid injection of kainic acid (0.1 microg) and were terminated after 45 min by diazepam (30 mg/kg) administration. Animals were killed 0-72 h following diazepam administration. Levels of the 32-kDa proenzyme form of caspase-3 were unaffected by seizures. Levels of the 17-kDa cleaved (active) fragment of caspase-3 were almost undetectable in control brain, but were increased significantly at 4 and 24 h within ipsilateral hippocampus and cortex in seizure animals. Caspase-3-like protease activity was increased within the ipsilateral hippocampus at 8 and 24 h following seizures. Caspase-3 immunoreactivity was increased within the vulnerable ipsilateral CA3/CA4 subfield at 24 and 72 h following seizures and was associated predominantly, but not exclusively, with neurons exhibiting DNA fragmentation. The putatively selective caspase-3 inhibitor N-benzyloxycarbonyl Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone significantly improved neuronal survival bilaterally within the hippocampal CA3/CA4 subfields following seizures. Collectively, these data suggest that caspase-3 may play a significant role in the mechanism by which neurons die following seizures. PMID- 10693955 TI - Early leptin response to a palatable diet predicts dietary obesity in rats: key role of melanocortin-4 receptors in the ventromedial hypothalamic nucleus. AB - We have investigated whether interactions between leptin and hypothalamic melanocortin-4 receptors (MC4-Rs) determine individual susceptibility to dietary obesity in rats. Animals with relatively high plasma leptin levels 1 week after presentation of palatable food, before weight increased significantly, subsequently showed lower food intake and weight gain after 8 weeks of palatable feeding than those with low early leptin levels. The rats with lesser weight gain also showed significantly greater down-regulation of MC4-Rs, which mediate hypophagia, in specific hypothalamic areas, namely, the arcuate, dorsomedial, and ventromedial (VMH) nuclei and the median eminence. We suggest that this reflects enhanced receptor exposure to endogenous alpha-melanocyte-stimulating hormone, an appetite-suppressing peptide produced by hypothalamic proopiomelanocortin neurones. It is striking that plasma leptin levels at 1 week were inversely correlated with MC4-R density in the VMH, suggesting that this is a key site of leptin action. The early leptin response to palatable feeding may therefore "program" subsequent feeding behaviour and weight gain by regulating neurones that project selectively to the VMH. PMID- 10693956 TI - L-DOPA does not enhance hydroxyl radical formation in the nigrostriatal dopamine system of rats with a unilateral 6-hydroxydopamine lesion. AB - The debate about the toxicity of L-DOPA to dopaminergic neurons has not been resolved. Even though enzymatic and nonenzymatic metabolism of L-DOPA can produce hydrogen peroxide and oxygen free radicals, there has been controversy as to whether L-DOPA generates an oxidant stress in vivo. This study determined whether acute or repeated administration of L-DOPA caused in vivo production of hydroxyl radicals in striatum and other brain regions in rats with a unilateral 6 hydroxydopamine lesion of the dopaminergic nigrostriatal projections. Salicylate trapping combined with in vivo microdialysis provided measurements of extracellular 2,3-dihydroxybenzoic acid (2,3-DHBA) in striatum following L-DOPA administration systemically (100 mg/kg, i.p.) or by intrastriatal perfusion (1 mM, via the microdialysis probe). Tissue concentrations of 2,3-DHBA and salicylate were also measured in striatum, ventral midbrain, and cerebellum following repeated administration of L-DOPA (50 mg/kg, i.p., once daily for 16 days). In each instance, treatment with L-DOPA did not increase 2,3-DHBA concentrations, regardless of the nigrostriatal dopamine system's integrity. When added to the microdialysis perfusion medium, L-DOPA resulted in a significant decrease in the striatal extracellular concentration of 2,3-DHBA. These results suggest that administration of L-DOPA, even at high doses, does not induce hydroxyl radical formation in vivo and under some conditions may actually diminish hydroxyl radical activity. Furthermore, prior damage to the nigrostriatal dopamine system does not appear to predispose surviving dopaminergic neurons to increased hydroxyl radical formation following L-DOPA administration. Unlike L-DOPA, systemic administration of methamphetamine (10 mg/kg, s.c.) produced a significant increase in the concentration of 2,3-DHBA in striatal dialysate, suggesting that increased formation of hydroxyl radicals may contribute to methamphetamine neurotoxicity. PMID- 10693957 TI - Functional expression of the serotonin transporter in immortalized rat brain microvessel endothelial cells. AB - There is evidence from recent studies that the brain endothelium (of capillaries and/or larger vessels) may serve as a specific target for serotonin [5 hydroxytryptamine (5-HT)]. This neurotransmitter is expected to be involved in the regulation of the blood-brain barrier (BBB) permeability and/or of the cerebral blood flow via receptor-mediated mechanisms. Effective control of these processes depends on a speedy uptake and metabolism of released 5-HT molecules. To realize this, a similar mechanism of 5-HT uptake as in brain may exist at the BBB. In this study, we have demonstrated using RT-PCR that 5-HT transporter mRNA is present in the brain endothelium and that a saturable transport system for 5 HT is functionally expressed in immortalized rat brain endothelial cells (RBE4 cells). These cells take up [3H]5-HT by an active saturable process with a Km value of 397 +/- 64 nmol/L and a transport capacity of 51.7 +/- 3.5 pmol x g(-1) x min(-1). The 5-HT uptake depends on Na+, as indicated by the replacement of NaCl by LiCl. The 5-HT uptake was sensitive to specific 5-HT transport inhibitors such as paroxetine, clomipramine, fluoxetine, and citalopram but not to inhibitors of the vesicular amine transporter such as reserpine or tetrabenazine. Our results demonstrate that cerebral endothelial cells are able to participate actively in the removal and metabolism of the released 5-HT, which supports the concept of direct serotoninergic regulation of the BBB function. PMID- 10693958 TI - ATP-sensitive potassium channel regulates astrocytic gap junction permeability by a Ca2+-independent mechanism. AB - Using the scrape-loading technique in cultured astrocytes, we show that sulfonylureas such as tolbutamide and glybenzcyclamide, which inhibit the ATP sensitive K+ channel, prevent the inhibition of gap junction permeability caused by several structurally unrelated uncouplers such as oleic acid, arachidonic acid, endothelin-1, octanol, and alpha-glycyrrhetinic acid. When the intracellular level of Ca2+ was diminished, all the uncouplers tested were still able to inhibit gap junction communication, indicating that their inhibitory effect was not mediated by Ca2+. In addition, tolbutamide and glybenzcyclamide prevented the inhibitory effect of these uncouplers in Ca(2+)-depleted astrocytes, suggesting that the inhibition of the ATP-sensitive K+ channel increases gap junction permeability through a Ca(2+)-independent mechanism. The activation of the ATP-sensitive K+ channel caused by potassium channel openers such as diazoxide and pinacidil led to the inhibition of gap junction communication and overcame the effect of sulfonylureas. These results suggest that the ATP-sensitive K+ channel regulates gap junctional permeability. PMID- 10693959 TI - Effects of repeated antidepressant treatment of type 4A phosphodiesterase (PDE4A) in rat brain. AB - In a previous study, an up-regulation of rolipram-sensitive, low-Km, cyclic AMP phosphodiesterase (PDE4) subtype PDE4A in rat cerebral cortex following repeated treatment of desipramine was observed. To determine whether this effect is shared by antidepressants from different pharmacological classes, PDE4A expression was examined using immunoblot analyses following repeated treatment with the norepinephrine re-uptake inhibitor desipramine, the monoamine oxidase inhibitor phenelzine, the atypical antidepressant trazodone, and the serotonin reuptake inhibitor fluoxetine. Desipramine, phenelzine, and fluoxetine all increased the intensities of the PDE4A bands in hippocampal preparations; trazodone did not. In preparations of cerebral cortex, the intensities of the PDE4A bands were increased following desipramine treatment, not changed following phenelzine or fluoxetine treatment, and decreased following trazodone treatment. It appears that repeated treatment with antidepressant drugs from different pharmacological classes produces similar effects on the expressions of PDE4A variants in hippocampus. This effect is not correlated with the changes in beta-adrenergic receptor densities, suggesting these antidepressants may at some point alter intracellular signal transduction pathways in a similar manner. PMID- 10693960 TI - Ascorbate inhibits edema in brain slices. AB - Ascorbate is an essential antioxidant in the CNS, localized predominantly in neuronal cytosol. Slices of mammalian brain rapidly lose ascorbate, however, when incubated in ascorbate-free media; brain slices also take up water and swell. Here we investigated water gain in coronal slices of rat forebrain incubated with and without ascorbate for 1-3 h at 34 degrees C. Slices progressively gained water in ascorbate-free media, with a significant 12% water increase after 3 h at 34 degrees C, compared with the water content of slices after a 1-h recovery period at 24 degrees C, immediately following slice preparation. Inclusion of 400 micro M ascorbate in the medium led to an increase in tissue ascorbate content and prevented water gain at 34 degrees C. By contrast, water gain was not inhibited by isoascorbate or thiourea, which are antioxidants that are not accumulated in brain cells. The oxidant H2O2 enhanced water gain, whereas a cocktail of NMDA and non-NMDA receptor blockers inhibited edema formation to the same extent as ascorbate. These data demonstrate that brain edema, linked to glutamate-receptor activation, can result from intracellular oxidative stress and that this can be prevented by ascorbate. PMID- 10693961 TI - Differential control of tyrosine hydroxylase activation and catecholamine secretion by voltage-operated Ca2+ channels in bovine chromaffin cells. AB - Contributions of L-, N-, and P/Q-type voltage-operated Ca2+ channels to two responses of bovine adrenal chromaffin cells have been studied using the nonreceptor stimulus K+ depolarization. Tyrosine hydroxylase activity and catecholamine secretion were both increased by K+ over a similar concentration range and in a Ca(2+)-dependent manner. At a submaximal concentration of 20 mM K+, tyrosine hydroxylase activation was reduced by nitrendipine but unaffected individually by (+/-)-Bay K 8644, omega-conotoxin GVIA, omega-agatoxin IVA, and omega-conotoxin MVIIC. It was fully blocked by combined inhibition of L-, N-, and P/Q-type channels. With a maximal concentration of 50 mM K+, tyrosine hydroxylase activation was unaffected by nitrendipine as well as by each of the other drugs on its own; however, it was reduced by 71 % by combined inhibition of L-, N-, and P/Q-type channels. In contrast, catecholamine secretion with both 20 and 50 mM K+ was enhanced by (+/-)-Bay K 8644, partially inhibited by nitrendipine and omega conotoxin MVIIC, and completely blocked by a combination of antagonists for L-, N , and P/Q-type channels. The results show that Ca2+ entry through voltage operated Ca2+ channels can differentially regulate distinct chromaffin cell responses and that this is an intrinsic property of the mechanisms by which Ca2+ entry activates these responses. It is not dependent on the parallel activation of other signaling events by receptors. PMID- 10693962 TI - Recombinant expression of alpha-bungarotoxin in Pichia pastoris facilitates identification of mutant toxins engineered to recognize neuronal nicotinic acetylcholine receptors. AB - A snake venom-derived alpha-neurotoxin, alpha-bungarotoxin (alphaBgtx), is the classic competitive antagonist of nicotinic acetylcholine receptors (nAChRs). The very high specificity and essentially irreversible binding of alphaBgtx to various nAChRs make alphaBgtx the prime candidate for studying the molecular determinants of specificity for nAChR-ligand interactions. To facilitate site directed mutagenesis of alphaBgtx for functional analysis, we have developed a recombinant expression system for alphaBgtx using the methylotropic yeast Pichia pastoris. A synthetic gene coding for alphaBgtx was subcloned into an expression vector that directs secretion of the recombinant alphaBgtx (rBgtx) when stably integrated into the yeast genome. Expression of rBgtx was induced by growth of yeast cultures with methanol as the sole carbon source. The activity of the rBgtx in the cell-free medium was measured by competition with 1251-Bgtx for binding to Torpedo nAChR-enriched membranes. The rBgtx, purified to homogeneity by standard HPLC, has the correct predicted amino terminal sequence and molecular mass. Its circular dichroism spectrum is very similar to that of authentic venom-derived alphaBgtx, and the biological activity of the rBgtx is identical to that of authentic alphaBgtx. We have used the Pichia expression system to study a double point mutation of alphaBgtx, rBgtx-K38P/L42Q, that has a high affinity for alpha3beta2 neuronal nAChRs. This is the first demonstration of engineering an alpha-neurotoxin to recognize non-alpha7 neuronal nicotinic receptors. PMID- 10693963 TI - Altered G protein-coupling functions of RNA editing isoform and splicing variant serotonin2C receptors. AB - Different isoforms of serotonin subtype 2C receptor (5-HT(2C)R) with altered G protein-coupling efficacy are generated by RNA editing, which converts genomically encoded adenosine residues into inosines. In combination, editing of five sites all located within the second intracellular loop region of 5-HT(2C)R mRNA changes the gene-encoded Ile, Asn, and Ile at positions 156, 158, and 160, respectively. We analyzed the G protein-coupling functions of previously unreported editing isoform receptors. An approximately 13-fold reduction in the agonist potency for G protein-coupling stimulation as well as a significantly reduced basal level activity was observed with the thalamus-specific isoform carrying Ile156, Gly158, and Val160 (5-HT(2C)R-IGV). In contrast, the agonist was four- to five-fold less potent with 5-HT(2C)R-MSV and -IDV, detected in the amygdala and choroid plexus, respectively, indicating a dominant role for the amino acid residue at position 158 in receptor functions. We also identified a splicing variant receptor with a truncated C terminus that displayed no ligand binding capacity or G protein-coupling activity. Examination of the alternatively spliced RNA encoding this truncated receptor suggests that editing of this variant RNA occurs after completion of splicing, resulting in complete editing at all five sites. PMID- 10693964 TI - Immunolocalization of the mGluR1b splice variant of the metabotropic glutamate receptor 1 at parallel fiber-Purkinje cell synapses in the rat cerebellar cortex. AB - Several metabotropic glutamate receptor (mGluR) subtypes have been identified in the cerebellar cortex that are targeted to different compartments in cerebellar cells. In this study, preembedding immunocytochemical methods for electron microscopy were used to investigate the subcellular distribution of the mGluR1b splice variant in the rat cerebellar cortex. Dendritic spines of Purkinje cells receiving parallel fiber synaptic terminals were immunoreactive for mGluR1b. With a preembedding immunogold method, approximately 25% of the mGluR1b immunolabeling was observed perisynaptically within 60 nm from the edge of the postsynaptic densities. Values of extrasynaptic gold particles beyond the first 60 nm were maintained at between 10 and 18% along the whole intracellular surface of the dendritic spine membranes of Purkinje cells. For comparison, the distribution of mGluR1a was studied. A predominant (approximately 37%) perisynaptic localization of mGluR1a was seen in dendritic spines of Purkinje cells, dropping the extrasynaptic labeling to 15% in the 60-120-nm bin from the edge of the postsynaptic specialization. Our results reveal that mGluR1b and mGluR1a are localized to the same subcellular compartments in Purkinje cells but that the densities of the perisynaptic and extrasynaptic pools were different for both isoforms. The compartmentalization of mGluR1b and mGluR1a might serve distinct requirements in cerebellar neurotransmission. PMID- 10693965 TI - Identification of amino acid residues of GABA(A) receptor subunits contributing to the formation and affinity of the tert-butylbicyclophosphorothionate binding site. AB - A chimeric GABA(A) receptor subunit was constructed that contained the beta3 sequence from the N-terminus to the first two amino acids of the second transmembrane (TM2) domain. The remaining part of this chimera had the sequence of the alpha1 subunit. On co-expression with alpha1 subunits, this chimera was able to form heterooligomeric channels that were open in the absence of GABA. Picrotoxin and tert-butylbicyclophosphorothionate (TBPS) were able to block these channels with low potency. These channels exhibited high-affinity [3H]muscimol but no high-affinity [35S]TBPS binding sites. Introduction of V251, A252, and L253 of the beta3 subunit into the chimera resulted in the formation of closed channels that could be opened by GABA. The introduction of A252 and L253 of the beta3 subunit into this chimera was sufficient to reconstitute the specific high affinity [35S]TBPS binding site in receptors composed of the chimera and alpha1 subunits. Replacement of other amino acids of the TM2 region of the chimera with corresponding amino acids of the beta3 subunit modulated the affinity of this [35S]TBPS binding site. Results obtained provide important information on the structure-function relationship of GABA(A) receptors. PMID- 10693966 TI - Characterization of carrier-mediated efflux in human embryonic kidney 293 cells stably expressing the rat serotonin transporter: a superfusion study. AB - Human embryonic kidney 293 cells stably transfected with the rat plasmalemmal serotonin transporter (rSERT) were incubated with 5-[3H]hydroxytryptamine ([3H]5 HT) and superfused. Substrates of the rSERT, such as p-chloroamphetamine (PCA) or methylenedioxymethamphetamine, concentration-dependently increased basal efflux of [3H]5-HT. 5-HT reuptake blockers (e.g., imipramine, citalopram) also caused an enhancement of [3H]5-HT efflux, reaching about half the maximal effect of the rSERT substrates. In uptake experiments, both groups of substances concentration dependently inhibited 5-HT uptake. EC50 values obtained in superfusion experiments significantly correlated with IC50 values from uptake studies (r2 = 0.92). Addition of the Na+,K(+)-ATPase inhibitor ouabain (100 microM) to or the omission of K+ from the superfusion buffer accelerated basal efflux. The effect of PCA (10 microM) was markedly enhanced by both measures, whereas the effect of uptake inhibitors remained unchanged. When [3H]MPP+, a substrate with low affinity for the rSERT, was used instead of [3H]5-HT for labeling the cells, uptake inhibitors failed to augment efflux. By contrast, PCA accelerated [3H]MPP+ efflux, and its effect was strongly enhanced in the presence of ouabain. The results suggest that the [3H]5-HT efflux caused by substrates of rSERT is carrier mediated, whereas efflux induced by uptake inhibitors is a consequence of interrupted high-affinity reuptake that is ongoing even under superfusion conditions. PMID- 10693967 TI - Angiotensin II modulates the activity of Na+,K+-ATPase in cultured rat astrocytes via the AT1 receptor and protein kinase C-delta activation. AB - In astrocytes the activity of the Na+,K(+)-ATPase pump maintains an inwardly directed electrochemical sodium gradient used by the Na+-dependent transporters and regulates the extracellular K+ concentration essential for neuronal excitability. We show here that incubation of cultured rat astrocytes with angiotensin II (Ang II) modulates Na+,K(+)-ATPase activity, in a dose- and time dependent manner. Na+,K(+)-ATPase activation was mediated by binding of Ang II to AT1 receptors as it was completely blocked by DuP 753, a specific AT1 receptor subtype antagonist. Stimulation of Na+,K(+)-ATPase activity by Ang II was dependent on protein kinase C (PKC) activation because PKC antagonists abolished the inducing effect of Ang II and the PKC activator phorbol 12-myristate 13 acetate enhanced transporter activity. Ang II stimulated translocation of PKC delta but not that of other PKC isoforms from the cytosol to the plasma membrane. These results indicate that the activity of Na+,K(+)-ATPase in astrocytes is increased by physiological concentrations of Ang II and that the AT1 receptor subtype mediates the Na+,K(+)-ATPase response to Ang II via PKC-delta activation. PMID- 10693968 TI - The organophosphate sarin, at low concentrations, inhibits the evoked release of GABA in rat hippocampal slices. AB - In the present study, the whole-cell mode of the patch-clamp technique was applied to neurons of the CA1 pyramidal layer of rat hippocampal slices to investigate the effects of the organophosphate (OP) sarin on field stimulation evoked and on tetrodotoxin (TTX)-insensitive postsynaptic currents (PSCs) mediated by activation of type A gamma-aminobutyric acid (GABA) receptors or AMPA type glutamate receptors. At 0.3-1 nM, sarin reduced the amplitude of GABA mediated PSCs and had no effect on the amplitude of glutamatergic PSCs evoked by field stimulation of neurons synaptically connected to the neuron under study. The effect of sarin on evoked GABAergic PSCs was unrelated to cholinesterase inhibition, was partially reversed upon washing of the neurons with sarin-free external solution, and was mediated by a direct interaction of the OP with muscarinic acetylcholine receptors present on presynaptic GABAergic neurons. Sarin had no effect on the amplitude or kinetics of GABA- or glutamate-mediated miniature postsynaptic currents (MPSCs) recorded in the presence of the Na+ channel blocker TTX (300 nM), indicating that the OP does not interact with GABA(A) or glutamate receptors. Further, sarin did not alter the frequency of GABAergic or glutamatergic MPSCs, a finding that led to the conclusion that this OP does not affect the TTX-insensitive release of neurotransmitters. A selective reduction by sarin of the action potential-dependent release of GABA in the hippocampus can account for the occurrence of seizures in intoxicated subjects. PMID- 10693969 TI - Cisplatin-induced DNA-platination in experimental dorsal root ganglia neuronopathy. AB - The mechanism(s) and site(s) of the neurotoxic effect of cisplatin (CDDP) are still not entirely elucidated. A more detailed knowledge of these aspects of CDDP treatment might be useful to obtain a better understanding of the pathogenesis of its peripheral neurotoxicity, which is the dose-limiting side effect of CDDP. In the present study, the occurrence of CDDP-induced DNA-platination in dorsal root ganglia (DRG) of rats was evaluated in relation to DRG neuron pathological changes and CDDP-induced neuronopathy. Eight adult Wistar rats were treated with 2 mg/kg i.p. CDDP twice weekly for 9 times to induce sensory peripheral neuropathy. DNA-platination in specimens of DRG and kidney was measured immunohistochemically, with a polyclonal antibody (GPt) detecting CDDP-induced Pt DNA adducts. Results were compared with those of untreated rats. Chronic CDDP induced neurotoxicity, in a well described experimental model of chronic CDDP neurotoxicity in the Wistar rat, was confirmed by sensory DRG neuronopathy with secondary neuropathy, and demonstrated by reduced pain detection, decreased nerve conduction velocity in the tail nerve as well as morphological and morphometric changes in DRG neurons. Nuclear immunostaining for Pt-DNA adducts was observed in tubular cells of the kidney in 75% of the evaluated CDDP-treated rats, while in DRG cells CDDP-induced Pt-DNA adducts formation was found in 43% of the evaluated CDDP-treated rats. CDDP-induced DNA-platination was demonstrated in rat DRG neurons using a schedule of chronic CDDP administration which induced the onset of a sensory neuronopathy with secondary peripheral neuropathy. This finding further supports the hypothesis that CDDP is neurotoxic because it directly damages the DRG neurons. PMID- 10693970 TI - Neurotoxicity evaluation of rats after subchronic inhalation exposure to isobutanol. AB - The subchronic neurotoxic effects of isobutanol were studied by exposing Sprague Dawley rats to isobutanol vapor concentrations of 0, 250, 1000, and 2500 ppm for 6 hrs/day, 5 days/wk, for 3 months. A comprehensive set of neurotoxicity tests (functional observational battery, motor activity, perfusion fixation neuropathology, and schedule-controlled operant behavior) including an assessment of complex behavior dependent on learning and memory was conducted. In addition, full histopathology and blood chemistry evaluations were conducted in order to assess any potential functional/behavioral effects in the context of other possible systemic toxicities. There were no morphological or behavioral effects indicative of a specific, persistent or progressive effect of isobutanol on the nervous system at exposure concentrations up to 2500 ppm. A slight decrease in response to external stimuli was observed during exposures at all concentrations. These effects are likely transient effects of acute exposure to isobutanol. PMID- 10693971 TI - Increase in signal intensities on T1-weighted magnetic resonance images in asymptomatic manganese-exposed workers. AB - OBJECTIVES: To clarify the clinical significance of increased signal intensities on T1 weighted magnetic resonance imaging (MRI) we performed a large-scale epidemiological study on asymptomatic manganese (Mn)-exposed workers with its focus on MRI. METHODS: We randomly selected 121 male workers out of a total of 750 workers including Mn-exposed, non-exposed manual, and non-exposed clerical workers in the factories. We studied environmental and biological monitoring, neurological examination, and MRI. RESULTS: The proportion of workers with increased signal intensities among the exposed, the non-exposed manual workers, and the non-exposed clerical workers was 46.1%, 18.8%, and 0%, respectively. Especially, 73.5% of the welders showed increased signal intensities. In no subject, were clinical signs of manganism observed. The pallidal index correlated with blood Mn concentration. CONCLUSION: Increase in signal intensities on the T1 weighted image reflect recent exposure to Mn, but not necessarily manganism. At which increase of signal intensity, the progression of manganism from Mn exposure occurs, remains to be solved. PMID- 10693972 TI - Brevetoxin modulates neuronal sodium channels in two cell lines derived from rat brain. AB - Single Na+ channel currents were recorded from cell-attached membrane patches from two neuronal cell lines derived from rat brain, B50 and B104, and compared before and after exposure of the cells to purified brevetoxin, PbTx-3. B50 and B104 Na+ channels usually exhibited fast activation and inactivation as is typical of TTX-sensitive Na+ channels. PbTx-3 modified channel gating in both cell lines. PbTx-3 caused (1) significant increases in the frequency of channel reopening, indicating a slowing of channel inactivation, (2) a change in the voltage dependence of the channels, promoting channel opening during steady-state voltage clamp of the membrane at voltages throughout the activation range of Na+ currents, but notably near the resting potential of these cells (-60 - -50 mV), and (3) a significant, 6.7 mV hyperpolarized shift in the threshold potential for channel opening. Na+ channel slope conductance did not change in PbTx-3-exposed B50 and B104 neurons. These effects of Pbx-3 may cause hyperexcitability as well as inhibitory effects in intact brain. PMID- 10693973 TI - The effects of postweaning low-level Pb exposure on sustained attention: a study of target densities, stimulus presentation rate, and stimulus predictability. AB - It has been suggested that behavioral effects associated with low-level lead (Pb) exposure are related to alterations in attentional processes. These "attention deficits" remain undefined, and few studies have directly addressed the effects of Pb on specific measures of attention. The current study assessed the impact of Pb on sustained attention, as one of the 3 primary clinical symptom domains of attention deficit disorder. Groups of rats were exposed from weaning (21 days of age) to either 0, 50, or 150 ppm Pb acetate in drinking water producing blood Pb levels of <5, 16.0, and 28.0 microg/dl, respectively. Rats were trained to discriminate a pulsing (target) from a non-pulsing (distracter) light. Parametric manipulations of target probability (10%-80%) and inter-stimulus interval (1 s-6 s, and variable) were imposed to evaluate whether effects of Pb were dependent upon attention demand. Sensitivity of the paradigm to changes in CNS function was validated by examining the effects of d-amphetamine (0.0-2.0 mg/kg) in a separate group of rats. Exposure to 50 ppm increased errors of omission only during sessions with long and variable inter-stimulus intervals. The 150 ppm group showed a small increase in errors of commission during sessions with a long inter stimulus interval. No other Pb-related effects were observed. The absence of any pronounced Pb effects in this study on this sustained attention task could indicate that: 1) any Pb-induced attention deficits are modified by reinforcement and time-out contingencies; 2) Pb disrupts other aspects of attention such as inability to tolerate delays or impulsivity; and 3) Pb-related deficits are due to other behavioral dysfunctions. PMID- 10693974 TI - Neurotoxicity of isoniazid and its metabolites in cultures of mouse dorsal root ganglion neurons and hybrid neuronal cell line. AB - Isoniazid (INH) is one of the anti-tuberculosis drugs widely prescribed for patients since the early 1950s. It is relatively nontoxic but some patients develop peripheral neuropathy attributed to a disturbance of vitamin B6 metabolism. Some isoniazid metabolites are hepatotoxic but little is known about their neurotoxic property. Isoniazid and its metabolites including acetylisoniazid, acetylhydrazine, diacetylhydrazine, isonicotinic acid and hydrazine were examined for their potential neurotoxic effects in cultured mouse dorsal root ganglion (DRG) neurons and mouse neuroblastoma x DRG neuron hybrid cell line N18D3. Isoniazid did not cause neurotoxicity at exposures up to 7 days. Hydrazine was found to be the most toxic metabolite with LC50 values of 2.7 mM and 0.3 mM after 7 days of exposure in DRG neurons and N18D3 hybrid neurons, respectively. Other metabolites including acetylisoniazid, acetylhydrazine, diacetylhydrazine and isonicotinic acid had moderate to minor neurotoxic effects on N18D3 hybrid neurons. Pyridoxine, which is used in clinical practice to prevent or ameliorate the isoniazid-induced neuropathy, did not consistently reverse the neurotoxicity of any of the metabolites in the cell cultures, but some interaction with hydrazine cannot be ruled out. Pyridoxine itself was found to be neurotoxic both in DRG neurons and N18D3 hybrid neurons, in agreement with human peripheral sensory neuropathy caused by prolonged overdosage. The enzymes catalase and superoxide dismutase and the antioxidant agent selenium showed some protection against hydrazine neurotoxicity, suggesting an involvement of the generation of reactive oxygen species in the pathogenesis of isoniazid neuropathy. Both mouse DRG neurons and N18D3 mouse hybrid neurons were shown to be useful culture systems for elucidating the neurotoxicity mechanisms of agents causing sensory neuropathies and general neurotoxic effects in the nervous system. PMID- 10693975 TI - Tremor frequency patterns in mercury vapor exposure, compared with early Parkinson's disease and essential tremor. AB - A new portable tremometer allows determination of tremor intensities at different tremor frequencies. Based on past studies, two tremor frequency windows of similar size were chosen at 3.0-6.5 Hz and 6.6-10.0 Hz to reflect major tremor intensities in Parkinson's disease and mercury vapor poisoning, respectively. In 81 healthy controls, total tremor intensity was higher for the preferred hand and depended on age. Ten patients treated for Parkinson's disease showed substantially increased tremor intensity, especially within the low-frequency window. This pattern was also apparent in 14 patients with de novo Parkinson's disease whose overall tremor intensity was only mildly elevated. In contrast, ten patients with essential tremor had peak frequencies in both windows, and some patients had increased tremor on one side only. Sixty-three Brazilian gold traders exposed to mercury vapor showed increased tremor predominantly in the high-frequency window. Three of the gold traders had a narrower tremor peak at frequencies of 7-8 Hz. While the urine-mercury concentration was significantly associated with the current number of burning sessions per week, it did not correlate with tremor intensities. However, eight traders had a urinary mercury excretion level above 50 microg and at the same time a greatly increased average tremor intensity within the high-frequency window. These patterns were statistically significant for relative tremor intensities, but were less clear when total intensities were used. These observations suggest that the relative distribution of tremor intensities in specific frequency bands may be a valuable supplement to current diagnostic methods for subjects with mercury vapor exposure. PMID- 10693976 TI - Morphometric studies of myelination in the spinal cord of mice exposed developmentally to aluminum. AB - Swiss-Webster mice were exposed to diets containing 7 or 1000 microg aluminum (Al)/g as Al lactate from conception through maturity (45 days of age). This exposure has previously been shown to cause changes in CNS myelin composition and peroxidizability; in this study myelin sheath widths were measured. Initially, samples of epon embedded, toluidine blue stained cervical spinal cord sectioned at 0.5 mm were examined light microscopically. Qualitatively, Al-treated mice appeared to have a diffuse paleness in nerve tracts. No indication of myelin structural damage (splitting, degeneration) was noted. Quantitative microscopy was performed using images captured with Scion Image Dage 1.59 at 1000x with oil. Axon perimeters and sheaths were measured with NIH image using a standardized sampling pattern in the right medial dorsal and ventral regions of the cervical spinal cord in 6 mice (3 male, 3 female) per group. Mean myelin sheath widths were 16% smaller in the Al-treated group compared to controls (p=.03). There was no effect of sex or region (dorsal/ventral). Axon perimeters were also smaller on the average in the Al treated group but this difference was not significant (p=.16). The relationship between sheath width and axon diameter was similar in the two groups. The density of myelinated axons was greater in some areas for the Al-treated group. The data indicate that dietary aluminum exposure can interfere with myelination in the spinal cord. PMID- 10693977 TI - Aluminum uptake and effects on transferrin mediated iron uptake in primary cultures of rat neurons, astrocytes and oligodendrocytes. AB - Transferrin (Tf) is known primarily for its role in the transport and cellular uptake of iron (Fe). Tf is also the major serum binding protein for Al. In this study, primary rat oligodendrocyte, neuron and astrocyte cultures were found to differ in Tf mediated Fe and Al uptake and in the effect of Al-Tf on Fe-Tf uptake during 4 h incubation periods. When incubated with Al-Tf (1.25 microM), oligodendrocytes displayed a 3- to 4-fold increase (p=.0002) in Al, neurons demonstrated a much smaller (p=.06) increase, and no increase was seen for astrocytes. When incubated with equimolar Al citrate or Al chloride, no increase in cellular Al was seen in any of the three cell types. Oligodendrocytes, astrocytes and neurons all demonstrated greater 59Fe uptake from Fe-Tf than Fe chloride. This uptake could be inhibited by excess Fe-Tf in oligodendrocytes and neurons, but not astrocytes. A small but significant inhibition of 59Fe uptake from Fe-Tf was seen after addition of Al-Tf to the incubation medium of oligodendrocytes, but not neurons or astrocytes. Oligodendrocytes may be particularly vulnerable to the accumulation of excess intracellular Al, and to interference of Al with Fe uptake. Such effects could contribute to Al-induced neurotoxicity if they result in altered myelin formation or maintenance. PMID- 10693978 TI - Immunohistochemical study of phosphorylated neurofilaments during the evolution of organophosphorus ester-induced delayed neuropathy (OPIDN). AB - Organophosphorus ester-induced delayed neuropathy (OPIDN) is manifest by delayed degeneration of distal levels of long myelinated fibers following an appropriate neurotoxic exposure. We investigated the dynamics of cytoskeletal changes during nerve fiber degeneration in this condition, focusing on the immunohistochemistry of axonal phosphorylated neurofilaments. OPIDN was produced in 5-month-old White Leghorn hens using a single 2.5 mg/kg intramuscular dose of phenyl saligenin phosphate. Hens were sacrificed on days 4, 7, 9, 15, and 20, and the tibial nerve branch to the gastrocnemius muscle was studied by light microscopy and immunohistochemistry (using the SMI 31 monoclonal primary antibody to phosphorylated neurofilaments). At post-dosing days 9, 15, and 20 various stages of OPIDN lesions were noted, including axonal swelling and myelinated nerve fiber degeneration. These were associated with intra-axonal cytoskeletal lysis, manifest by loss of immunolabeled phosphorylated neurofilaments, a process consistent with proteolysis. Aggregations of excess axonal phosphorylated neurofilaments were not observed. PMID- 10693979 TI - Dose response of ethanol ingestion on antioxidant defense system in rat brain subcellular fractions. AB - This study investigated the response of the antioxidant defense system in brain subcellular fractions after oral graded doses of ethanol to rat. Four groups of male Fischer-344 rats were orally administered saline, ethanol 2 g, 4 g, and 6 g/kg, respectively, and sacrificed 1 hour post treatment. Brain cytosol, synaptosomes, microsomes and mitochondria were separated by density gradient differential centrifugation and assayed for antioxidant system. A significant and dose-dependent-decrease in superoxide dismutase (SOD) activity was observed in all brain subcellular fractions. Catalase (CAT) activity was significantly decreased in brain mitochondria (67% and 80% of control) at higher doses of ethanol; whereas, CAT activity was significantly increased in cytosol, synaptosomes and microsomes. Glutathione peroxidase (GSH-Px) activity was significantly increased in all brain subcellular fractions except in cytosol at higher dose of ethanol. Malondialdehyde (MDA) content was significantly increased in all brain subcellular fractions showing dose response of ethanol-induced oxidative stress. The increase in MDA levels in the brain synaptosomes and microsomes were higher at 6 g dose of ethanol (155% and 163% of control) when compared to mitochondria and cytosol. Glutathione (GSH) levels were significantly increased in brain cytosol and microsomes at higher dose of ethanol (164% and 159% of control); whereas, the GSH concentration was significantly decreased in brain synaptosomes and mitochondria. The antioxidant enzyme (AOE) activity ratios (GSH-Px/SOD and GSH-Px + CAT/SOD) were dose dependently increased in all brain subcellular fractions, particularly in synaptosomes. The GSH/GSSG ratio was dose dependently increased in brain microsomes. The perturbations in the antioxidant defense system and enhanced lipid peroxidation following graded doses of ethanol ingestion indicate a dose-dependent-oxidative 2133stress response in brain subcellular compartments of rats. PMID- 10693980 TI - Minor immunophilin binding of tacrolimus and sirolimus metabolites. AB - OBJECTIVES: We have previously identified three minor immunophilins of molecular weights 37 kDa, 14 kDa, and 5-8 kDa capable of binding tacrolimus and sirolimus. DESIGN AND METHODS: When tested against pure preparations of five sirolimus metabolites, the 14 kDa protein had almost no cross-reactivity, the 37 kDa protein cross-reacted from a high of 23.2% to <10% and the 5-8 kDa protein cross reacted from <10% to 46.4%. When the 5-8 kDa immunophilin was tested in whole blood samples to assess interference in clinical samples, the demethylated sirolimus metabolites showed about 25% less cross-reactivity while the hydroxylated metabolites reacted about the same. RESULTS: Since MLC data on sirolimus metabolites to date indicates that their pharmacologic potencies are < or =10% of the parent, the 14 kDa immunophilin appears to be the best candidate for a sirolimus radioreceptor assay. The 5-8 kDa immunophilin is newly identified and its cross-reactivity with tacrolimus metabolites had not been assessed. Binding of the 5-8 kDa immunophilin to pure preparations of three tacrolimus metabolites showed virtually no binding of the protein to 13-O-demethyl and 31-O demethyl tacrolimus and binding to 15-O-demethyl tacrolimus at 121% relative to parent tacrolimus. Cross-reactivity of 15-O-demethyl tacrolimus with the 5-8 kDa protein was then assessed in whole blood samples, and it bound at a level of 163%. MLC data indicates that 31-O-demethyl tacrolimus is equipotent to parent tacrolimus in immunosuppressive activity, while the 13-O-demethyl and 15-O demethyl have negligible immunosuppressive activity. CONCLUSIONS: Therefore, the 5-8 kDa immunophilin would have limitations in a radioreceptor assay for tacrolimus. In addition, we have evidence that the 5-8 kDa immunophilin is a subunit of a 52 kDa immunophilin previously identified by our group, and the cross-reactivity of the 5-8 kDa immunophilin with these metabolites is similar to that found previously with the 52 kDa, indicating that the two proteins could be related. PMID- 10693981 TI - Enzyme immunoassay for the measurement of human tenascin-C on the Bayer Immuno 1 analyzer. AB - OBJECTIVES: To evaluate a new tenascin-C assay performed on the Bayer Immuno 1 system. DESIGN AND METHODS: The precision was measured using three levels of serum pools. Linearity was tested by diluting patient serum samples containing high tenascin-C concentrations, and the minimal detectable concentration determined by repetitive analysis of the zero calibrator. Preliminary reference intervals were determined by testing serum samples from 220 healthy individuals. Biovariability was estimated in a cohort of 20 apparently healthy subjects over 18 days. The levels of tenascin-C in patients with different liver diseases was tested. RESULTS: The detection limit was 2 ng/mL. At concentrations ranging from 325 to 1957 ng/mL the assay demonstrated within-run and between-run CVs ranging from 4% to 3.6% and 8.4% to 6.7%, respectively. Dilutions of sera were linear and parallel to the standard curve with recoveries ranging from 97% to 100%. The reference interval (central 95% interval) for tenascin-C in serum of healthy adults was 199-906 ng/mL. The variability study yielded an analytical variability, CV(A), of 1.8%; a within-subject variability, CV(I), of 11.7%; and a between-subject variability, CV(G), of 39.3%. Tenascin-C concentrations in sera of liver disease patients were significantly increased. CONCLUSIONS: The novel assay provides a rapid and reliable procedure for the determination of tenascin-C levels in human sera. PMID- 10693982 TI - Laboratory standardization of a large international clinical trial: the DAIS experience. DAIS Project Group. Diabetes Atherosclerosis Intervention Study. AB - OBJECTIVE: To implement a quality control program for the standardization and harmonization of lipid and lipoprotein analyses as performed at two core laboratories (St. Paul's Hospital, UBC [Vancouver], and NPHI [Helsinki]) for the Diabetes Atherosclerosis Intervention Study (DAIS). DESIGN AND METHODS: A DAISSOFT computer program was designed to minimize the occurrence of data and sample management errors during the course of the study. Fresh human serum was used for the provision of an accuracy based external quality control program that monitored the analytical performance of lipid testing at these two laboratories. A separate program was designed for monitoring hemoglobin A1c (HbA1c). At the outset of the study, allowable total error goals were established for each analyte. Ongoing performance was monitored using bimonthly blinded challenges of fresh human serum. The two EQA programs routinely monitored the analysis of total cholesterol, calculated LDL-cholesterol, HDL-cholesterol, net triglycerides, apoprotein A-1, apoprotein B, and HbA1c. RESULTS: The EQA precision and accuracy data for the measurement of total cholesterol at the two core laboratories over the last 5 years indicated both laboratories operated with good precision, approximately 1% CV over the time period. The accuracy at both laboratories was similar initially. Part way through the study, the accuracy of the cholesterol method at NHPI tended to drift upward with an operating positive bias (+3%) relative to the Abell Kendall reference method. Triglyceride measurements were the most problematic for the study. By EQA cycle 8, the accuracy of the method at UBC had stabilized and was meeting the accuracy goals of the study. NPHI's method was negatively biased relative to the accuracy base of the DAIS study. In spite of recalibrating their method, NPHI found it difficult to maintain consistent accuracy for the measurement of triglycerides during the study. Both laboratories operated their HDL methods with excellent precision. Accuracy at NHPI was well maintained over the course of the study whereas the accuracy of HDL measurements at UBC was more problematic. There was an inconsistent variation in the accuracy of apoprotein A-1 measurements at both laboratories. In most cases, the bias would be corrected by the time of the next EQA challenge. In the case of apo B, one laboratory was standardized to the CDC while the other laboratory was standardized to IFCC/WHO. The discrepancy between these two accuracy bases was >20%. Recalibration to a common accuracy base rectified the problem. Only minor problems were encountered with the precision and accuracy of the DIAMAT assay for hemoglobin A-1c. The two DAIS core laboratories consistently operated within the 9% total error goals of the study for HbA1c. CONCLUSIONS: Through the use of this program, the two DAIS core laboratories were able to maintain their lipid analyses within the limits of allowable total error that had been established for the study. PMID- 10693983 TI - Nonenzymatic elimination of ascorbic acid in clinical samples. AB - OBJECTIVES: Ascorbic acid interferes significantly in the oxidative reaction of chromogenic reagents by peroxidase and hydrogen peroxide. Currently, ascorbate oxidase is commonly utilized for eliminating the interference of ascorbic acid in the oxidative colorimetric reaction. This enzyme, however, displays several disadvantages, such as high cost, variation from lot to lot, and low stability. We applied a series of commercially available and stable radicals (ascorbic acid quenchers [AAQs]) for nonenzymatic quenching of ascorbic acid in the uricase based uric acid determination in serum and urine. DESIGN AND METHODS: In order to evaluate the quenching activity of AAQs, a commercially available uric acid detection kit was used. TBA-80FR.NEO biochemical analyzer was utilized for the assay. RESULTS: 4-Hydroxy-2,2,6,6-tetramethyl-1-piperidinyloxy free radical (AAQ 2) was the most effective ascorbic acid quencher among the four stable radicals, and the uric acid assay suffered no interference by AAQ-2. The ascorbic acid quenching ability of 2 mmol/L of AAQ-2 in reagent solution (reagent-I) was > or = 2 U/ml ascorbate oxidase in reagent solution. CONCLUSIONS: AAQ-2 was proven to be a suitable quencher of ascorbic acid in clinical samples. PMID- 10693984 TI - Comparison of steady-state trough sirolimus samples by HPLC and a radioreceptor assay. AB - OBJECTIVES: We have previously identified a minor immunophilin of 52 kDa molecular weight capable of binding tacrolimus and sirolimus. Because immunophilins are capable of binding both parent drug and metabolites and HPLC assays are typically used to assess parent drug in clinical situations, we used this immunophilin in a radioreceptor assay (RRA) to determine if any metabolites not included in the HPLC measurement would bind to the immunophilin and be associated with thrombocytopenia in patients receiving sirolimus. DESIGN AND METHODS: We tested 51 steady-state trough whole blood samples from non thrombocytopenic patients and 51 steady-state trough samples from thrombocytopenic patients and compared them to HPLC measurements of parent drug in the same samples. We also tested whole blood samples spiked with authentic sirolimus metabolites using RRA to ascertain the effect these metabolites have on the technique. RESULTS: We found minimal cross-reactivity in this assay for sirolimus metabolites (binding ranged from <10% to 26%), and good correlation of the radioreceptor assay with HPLC (linear regression slope 0.92, y-intercept 0.79). There was no statistically significant difference between the RRA and HPLC results in two patient groups-thrombocytopenic and non-thrombocytopenic-using the paired t-test (p<0.005) and Bland-Altman analysis. CONCLUSIONS: These findings indicate that although the RRA could be substituted for HPLC in therapeutic drug monitoring, the 52 kDa immunophilin does not offer an advantage in terms of detecting metabolites associated with thrombocytopenia. However, the RRA offers the advantages of shorter turnaround time, smaller sample volume and potential for automation. PMID- 10693985 TI - Plasma, urinary and fecal potassium changes in athletes during ambulatory, periodic, and continuous hypokinetic conditions. AB - OBJECTIVES: Prolonged hypokinesia (HK) induces significant electrolyte changes, but little is known about the effect of prolonged periodic hypokinesia on plasma, urinary, and fecal K. The aim of this study was to measure potassium (K) changes during prolonged periodic (PHK) and continuous (CHK). DESIGN AND METHODS: Studies were done during the pre HK and HK periods. Thirty male athletes were chosen as subjects. They were divided equally into three groups: unrestricted ambulatory control subjects (UACS), continuously hypokinetic subjects (CHKS), and periodically hypokinetic subjects (PHKS). The CHKS group was kept on a running distance of 0.7 km/day, while the PHKS group kept on a running distance of 0.7 and 11.7 km/day for 5 days and 2 days per week, respectively. The UACS group was on a running distance of 11.7 km/day. RESULTS: The following were measured: fecal K excretion; urinary K; sodium (Na) and chloride (CI) excretion; plasma K; Na and CI concentration; plasma renin activity (PRA) and plasma aldosterone (PA) concentration; physical characteristics; and peak oxygen uptake. Fecal K, urinary K, Na and CI excretion, plasma K, Na and CI concentration, and PRA and PA concentration, increased significantly (p< or =0.01) in the CHKS and PHKS groups when compared with the UACS group. Body weight and VO2 peak decreased significantly (p< or =0.01) in the CHKS group, while body weight increased and VO2 peak decreased significantly (p< or =0.01) in the PHKS group when compared with the UACS group. The measured parameters changed much more in the PHKS group than in the CHKS group. By contrast, the measured parameters did not change significantly in the UACS group when compared with the baseline control values. CONCLUSION: It was shown that prolonged PHK and CHK induce significant plasma and excretory K changes; however, plasma and excretory K changes were much greater in the PHKS group than in the CHKS group. It was concluded that the greater the stability of muscular activity, the smaller the plasma, urinary, and fecal K changes during prolonged HK. PMID- 10693986 TI - Serum 1,25 dihydroxy vitamin D (1,25(OH)2D3), 25 hydroxy vitamin D (25(OH)D) and parathormone levels in diabetic retinopathy. AB - OBJECTIVES: To investigate whether there is a relationship between serum 1,25 dihydroxy vitamin D3 [1,25(OH)2D3], which is an inhibitor of angiogenesis, concentrations and severity of diabetic retinopathy (DR). DESIGN AND METHODS: Serum 1,25(OH)2D3, 25 hydroxy vitamin D [25(OH)D] and parathormone (PTH) concentrations were measured in diabetic patients (n = 66) and nondiabetic healthy subjects (n = 20). RESULTS: The mean serum 1,25(OH)2D3 concentration in diabetic patients was lower than that in nondiabetics (57.3+/-21.44 vs. 89.4+/ 18.01 pmol/L, p<0.001); mean 1,25(OH)2D3 concentrations fell with increasing severity of DR [being 63.4+/-17.26 pmol/L for background DR (BDR), 47.7+/-13.27 pmol/L for preproliferative DR (pre-PDR), and 43.1+/-19.45 pmol/L for proliferative DR (PDR)]. Compared with the control group, serum 25(OH)D concentrations were found to be decreased in diabetic patients (p<0.001). There were negative correlations between 1,25(OH)2D3 and age (r = -0.331, p<0.01) and duration of diabetes (r = -0.255, p<0.05). CONCLUSION: From these findings, it was found that there was an inverse relationship between the severity of the retinopathy, i.e., neovascularization, and serum 1,25(OH)2D3 concentrations, being the lowest in PDR and the highest in diabetic patients without retinopathy (NDR) patients. The measurement of serum 1,25(OH)2D3 concentrations might be helpful to predict severity of DR in patients with diabetes mellitus. PMID- 10693987 TI - p53 gene mutation, tumor p53 protein overexpression, and serum p53 autoantibody generation in patients with breast cancer. AB - OBJECTIVES: Autoantibodies against the p53 tumor suppressor protein have been detected in the serum of a proportion of patients with various cancers. The generation of such antibodies has been proposed to be due to either tumor p53 protein accumulation or to the type of p53 gene mutation. These hypotheses are examined in the present study. DESIGN AND METHODS: Using immunofluorometric assays, we studied 195 patients with primary breast cancer for the presence of p53 antibodies in serum and p53 protein accumulation in the corresponding tumor. Seventeen patients (9%) were p53 antibody-positive and 77 (40%) overexpressed p53. Ten of the 17 p53 antibody-positive patients had tumor p53 accumulation and 7 were negative for p53. Statistical analysis revealed a weak association between the presence of p53 antibodies and p53 protein accumulation (p = 0.05). Direct DNA sequencing of exons 1-11 of the p53 gene was performed for 16 p53 antibody positive and 16 p53 antibody-negative patients. RESULTS: Five of the seropositive and eight of the seronegative patients had a p53 gene mutation. Four of the five mutations in the p53 antibody-positive patients affected a Tyr residue, whereas none of the gene abnormalities in the seronegative patients had such an effect. CONCLUSIONS: We conclude that p53 antibodies tend to develop in patients with tumor p53 accumulation, but p53 accumulation is neither sufficient nor necessary for the generation of the immune response. Further, p53 antibody-positive patients do not have higher frequency of p53 gene mutations than p53 antibody negative patients, but the former patient group is associated with a Tyr substitution in the protein product. PMID- 10693988 TI - Assay of creatinine using the peroxidase activity of copper-creatinine complexes. AB - OBJECTIVES: It was our goal to develop a urine dipstick that could measure creatinine with a peroxidase reaction. The simultaneous measurement of albumin and creatinine permits the estimation of the 24-h albumin excretion, an important value in judging existing or likely development of renal failure. A highly sensitive dye-binding dipstick method for albumin exists, and a suitable dipstick for the assay for urine creatinine is described here. METHODS: Copper-creatinine and iron-creatinine complexes have peroxidase activity. With 3,3',5,5' tetramethylbenzidine (TMB), and diisopropyl benzene dihydroperoxide (DBDH); the peroxidase activity of copper-creatinine and iron-creatinine complexes can be demonstrated. This reaction was used in the assay of urine creatinine either in solution or by a suitably impregnated urine dipstick. RESULTS: Our method based on the peroxidase activity of the copper-creatinine complex has an analytical range for creatinine of 100 mg/L (0.884 mmol/L) to 3000 mg/L (26.52 mmol/L). The creatinine assay is free from most interfering compounds that may be present in urine. Hemoglobin is an interferent, and its effects can be reduced but not eliminated by the addition of 4-hydroxy-2-methyl quinoline. We do not recommend using the dipsticks when visible blood is present or if the dipstick blood test is positive. The copper-creatinine complex oxidizes ascorbic acid; however, we were able to modify the reaction conditions so that ascorbic acid at < 4.4 g/L does not interfere. We found good agreement on fresh urines between the creatinine dipstick results and those by a standard rate-Jaffe cuvet method for creatinine. DISCUSSION: With the simultaneous measurement of creatinine and albumin in urine, the albumin/creatinine ratio can be determined effectively reducing or eliminating the occasional false-negative and false-positive result in those with dilute or concentrated urines, respectively. The dipstick test for these analytes permits the simple identification of individuals with possible albuminuria and could serve well in a point-of-care setting. PMID- 10693989 TI - Correlation between susceptibility of LDL subfractions to in vitro oxidation and in vivo oxidized LDL. PMID- 10693990 TI - Reference interval of ferritin in premenopausal women calculated in four laboratories using three different analyzers. PMID- 10693991 TI - Alpha1-antitrypsin deficiencies masked by a clinical capillary electrophoresis system (CZE 2000). PMID- 10693992 TI - Superantigens, conventional antigens and the etiology of Kawasaki syndrome. PMID- 10693993 TI - Bacteriologic and clinical efficacy of amoxicillin/clavulanate vs. azithromycin in acute otitis media. AB - OBJECTIVES: To compare the bacteriologic and clinical efficacy of amoxicillin/clavulanate and azithromycin in patients with acute otitis media (AOM), particularly the ability to eradicate the predominant AOM pathogens from middle ear fluid as assessed by mandatory second tympanocentesis. METHODS: In this single blind study 238 infants and children with AOM were randomized to receive amoxicillin/clavulanate (45/6.4 mg/kg/day in two divided doses for 10 days) or azithromycin (10 mg/kg on Day 1, then 5 mg/kg daily on Days 2 through 5). Tympanocentesis was performed before the first dose and repeated on Day 4, 5 or 6. Clinical response was assessed at end of therapy between Days 12 and 14 and at follow-up between Days 22 and 28. RESULTS: Amoxicillin/clavulanate was significantly more likely to eradicate all bacterial pathogens [83% (54 of 65) vs. 49% (35 of 71), P = 0.001] and Haemophilus influenzae [87% (26 of 30) vs. 39% (13 of 33), P = 0.0001] from middle ear fluid than was azithromycin. Amoxicillin/clavulanate was also more likely to eradicate Streptococcus pneumoniae, but the difference was not statistically significant [90% (18 of 20) vs. 68% (13 of 19) [corrected], P = 0.095]. On Days 12 to 14, signs and symptoms were more likely to resolve completely or improve in all culture-positive patients [86% (60 of 70) vs. 70% (51 of 73), P = 0.023] and in those with H. influenzae infections [91% (30 of 33) vs. 65% (22 of 34), P = 0.010] who received amoxicillin/clavulanate compared with those who received azithromycin. Otherwise there were no significant differences between groups in clinical outcomes on Days 12 to 14 or at follow-up. CONCLUSIONS: Our findings indicate that amoxicillin/clavulanate has superior bacteriologic and clinical efficacy compared with azithromycin in children with AOM. PMID- 10693994 TI - Testing for rubella-specific IgG antibody in urine. AB - BACKGROUND: In Japan rubella vaccination is generally done once during a lifetime, and the vaccination rate decreased after a revised vaccination law in 1995. History of rubella or vaccination may still be unreliable. Testing for rubella antibody is significant to prevent the occurrence of congenital rubella syndrome. However, the collection of blood samples to detect antibodies from young children is invasive and difficult. METHODS: For this study we obtained 853 matched serum and urine samples from 904 healthy students 10 or 14 years of age in the Ibara and Yoshii districts of Okayama, Japan, for a comparison of antibodies for rubella in the matched samples. The serum and urine antibodies were measured with hemagglutination-inhibition and enzyme-linked immunosorbent assays, respectively, and with our urine-based antibody test. RESULTS: The sensitivity, specificity and concordance rates of this urine-based antibody test were 96, 99 and 97% based on the serum antibody results of both assays. The coefficiency was 0.627 between the titers of the urinary and serum antibodies by the enzyme-linked immunosorbent assay. The urinary antibodies were stable for at least 5 months at 4 degrees C and 25 degrees C. CONCLUSIONS: Urine-based assay methods are helpful not only because they avoid the invasive approach of venipuncture but also because unprocessed urine specimens can be used and urinary antibody is stable for a long period. Therefore this test is suitable for screening. In addition protective amounts of rubella antibody in blood can be reliably assessed by means of urine samples. PMID- 10693995 TI - CD8+ T cell-mediated suppression of human immunodeficiency virus replication in older children with acquired immunodeficiency syndrome. AB - BACKGROUND: Suppression of HIV replication by CD8+ T cells and/or their products correlated with the survival of infants. We sought to elucidate the role of CD8+ T cell-mediated suppression in seven older children with AIDS. METHODS: After separation of each child's CD4+ and CD8+ T cells, three different HIV culture assays were performed: (1) patient CD4+ T cells and phytohemagglutinin (PHA) stimulated donor peripheral blood mononuclear cells (PBMC); (2) patient CD8+ T cells added to the CD4+ T cells and the PHA-stimulated donor PBMC (to test for CD8-mediated T cell suppression of HIV); (3) patient CD8+ cells added across a semipermeable membrane to the CD4+ T cells and the PHA-stimulated donor PBMC [to determine whether the CD8 cells secreted a soluble factor(s) that suppressed HIV]. RESULTS: Cultures from four of seven children showed greater HIV replication with CD4 cells alone than with CD4 and CD8 cells together, demonstrating CD8 suppression; evidence of soluble suppression was also seen. Cultures from two of the seven children showed HIV replication and no evidence of CD8 cell suppression. Cultures from one of the seven children had no appreciable replication of HIV even after removal of CD8 cells. CONCLUSIONS: CD8-mediated suppression is present in at least some children with AIDS. Additional mechanisms may be operating to slow the progression of the disease. PMID- 10693996 TI - Nasal quantity of respiratory syncytical virus correlates with disease severity in hospitalized infants. AB - OBJECTIVE: To evaluate the relationship between nasal quantity of respiratory syncytial virus (RSV) and disease severity in hospitalized infants without underlying cardiopulmonary disease or immunodeficiency. METHODS: Nasal aspirates were obtained from hospitalized infants <24 months of age with recently identified RSV infection and evaluated for RSV quantity by a standard plaque assay on HEp-2 cell monolayers. Subjects were classified as having "severe" disease if they required mechanical ventilation at the time of sample collection and as having "nonsevere" disease if they did not. Linear modeling was used to determine the relationship between nasal RSV quantity and various independent variables, including disease severity. RESULTS: Nasal aspirates from 39 patients were evaluated. Age, gender and mean duration of time from symptom onset to sample acquisition (5 days) were similar between the severe (n = 15) and nonsevere (n = 24) groups. Significantly more infants were born at <35 weeks gestation in the severe disease group (7 of 15 vs. 3 of 24, P = 0.017), and infants born at <35 weeks gestation were significantly more likely to be of non Caucasian ethnicity than were infants born at > or =35 weeks gestation (8 of 10 vs. 12 of 29, P = 0.035). The linear model found that higher nasal RSV quantities were associated with severe disease [mean +/- SEM, 5.06 +/- 0.34 log plaque forming units (pfu)/ml vs. 3.91 +/- 0.35 log pfu/ml, P = 0.022], gestational age > or =35 weeks (5.44 +/- 0.27 log pfu/ml vs. 3.52 +/- 0.45 log pfu/ml, P = 0.002) and non-Caucasian ethnicity (5.16 +/- 0.30 log pfu/ml vs. 3.80 +/- 0.37 log pfu/ml, P = 0.006). CONCLUSIONS: Nasal RSV quantity correlates with disease severity in hospitalized infants with recently identified RSV infection. PMID- 10693997 TI - Immunoblotting and serology for diagnosis of Helicobacter pylori infection in children. AB - BACKGROUND: The easiest way to identify the presence of current or past Helicobacter pylori infection is to test for antibodies. The aim of this study was to compare an enzyme-linked immunosorbent assay (ELISA) technique based on the detection of IgG antibodies directed against a global antigenic preparation with immunoblotting based on the analysis of IgG antibody reactivity to separate proteins. METHODS: Sera were collected from 80 children (mean age, 9.9 +/- 4.3 years). The reference tests were microbiologic and histologic examination of gastric biopsies obtained at upper endoscopy. RESULTS: The immunoblotting was more sensitive (100%) and specific (88%) than ELISA (96 and 79%, respectively) in the evaluation of H. pylori infection in children. Its positive predictive value was 92%, and its negative predictive value was 100%. The best performance index of immunoreactive bands to detect antibodies was obtained with the 26-kDa (88.7%), 30-kDa (77.5%) and 19.5-kDa (70%) antigens. Antibodies by immunoblot technique against the CagA antigen were present in 43.1% of children. CONCLUSION: Immunoblotting is highly sensitive and more specific than ELISA in children and provides additional information about the full serologic profile. Immunoblotting may therefore be a useful complement to serology, particularly in cases with doubtful ELISA results. PMID- 10693998 TI - Gram-negative bacillary bacteremia in human immunodeficiency virus type 1 infected children. AB - BACKGROUND: HIV-infected children are particularly susceptible to serious bacterial infections including Gram-negative bacillary bacteremia (GNB). However, the information available on GNB in these children is limited. METHODS: Retrospective review of hospital charts of HIV-infected children with GNB diagnosed between 1980 and 1997. The association between bacteremic episodes, degree of immunosuppression, HIV severity, medical treatment and clinical outcome was assessed. RESULTS: Of 680 HIV-infected children, 72 (10.6%) had 95 episodes of GNB. Statistical analyses were restricted to data from the first episode. The mean age (+/-SD) at diagnosis of GNB was 2.5 +/- 2.7 years (median, 1.6). The predominant organisms were Pseudomonas aeruginosa (26.4%), nontyphoidal Salmonella (15.3%), Escherichia coli (15.3%) and Haemophilus influenzae (12.5%). The relative frequency, per 5-year interval, of P. aeruginosa bacteremia steadily increased from 13% during 1980 through 1984 to 56% during 1995 through 1997. There were no cases of H. influenzae bacteremia after January 1, 1990. Eighty percent of GNB developed in children with AIDS and 72.2% developed in those with severe immunosuppression. Hypogamma-globulinemia and neutropenia were present in only 4.9 and 10.4% of first episodes, respectively. The overall case-fatality rate of GNB was 43.0%, and in children younger than 12 months it was 54.2%. CONCLUSIONS: A diagnosis of AIDS and/or severe immunosuppression was associated with increased risk of GNB, especially among younger children. Because of the high mortality of GNB, a broad spectrum antimicrobial therapy that effectively covers these organisms should be promptly instituted when bacteremia is suspected in HIV-infected children. PMID- 10693999 TI - Parents' preferences for outcomes associated with childhood vaccinations. AB - BACKGROUND: The number of shots in the childhood immunization schedule has been increasing and is likely to continue to increase in the coming years. Consideration of the psychologic costs of multiple injections, adverse events and vaccine-preventable disease is therefore growing in importance. METHODS: We assessed parent preferences, using both the time tradeoff (i.e. amount of parent time willing to trade) and willingness-to-pay (i.e. dollars willing to pay) metrics, for possible outcomes of vaccination among 206 parents of infants receiving care at Kaiser, Northern California Region. We also explored the relationship between preferences and subject characteristics. RESULTS: In general the amount of time subjects were willing to give up and the quantity of money they were willing to spend to avoid an outcome increased with the severity of the outcome. Preferences for our six main outcomes of interest all differed from one another (P < 0.0001, Tukey's multiple comparisons procedure). Rank correlation coefficients between time tradeoff and willingness-to-pay values for the six main outcomes ranged from 0.42 to 0.52 (all P < 0.004). Subject characteristics, including education, income, race/ethnicity and the child's birth order, did not explain the variation in parent preferences. CONCLUSIONS: In general subjects were willing to give up more money or time to avoid less desired outcomes. They were willing to give up only very small amounts of their own life expectancy or money to avoid minor, temporary outcomes (e.g. moderate fussiness, fever and pain) whereas they were willing to forego substantial lengths of their life or amounts of money to avoid a major, permanent outcome (i.e. permanent disability). Nonetheless much variation surfaced in the amount of time (or money) subjects were willing to trade to avoid outcomes. If this variation represents true differences in preferences, guideline developers must consider the role of individual parent preferences in decisions concerning vaccination. PMID- 10694000 TI - Human antibodies to pneumococcal surface protein A in health and disease. AB - BACKGROUND: Diseases caused by Streptococcus pneumoniae have a high impact in young children whose ability to mount antibodies to capsular polysaccharides is impaired. Pneumococcal surface protein A (PspA) is a potential vaccine candidate for this age group. METHODS: We used Western blot analysis and enzyme immunoassay to study human sera of healthy adults from Alabama (n = 20) and from Finland (n = 21), healthy children from Finland (n = 20) and ill children from Finland, those with pneumococcal invasive infection (n = 26) and those with nonpneumococcal invasive infection (n = 26). RESULTS: Human antibodies to PspA exhibited strong cross-reactivity among different pneumococcal strains. The geometric mean titer of IgG antibody to PspA in sera from 21 healthy adults was 4,040, from ten 3-year old healthy children 1,080 and from ten 2-month-old healthy children 1,650. The geometric mean titer of PspA antibody of acute phase sera of children with invasive pneumococcal disease was 140, significantly (P < 0.001) lower than the respective value, 1,020, for children with infection caused by other bacteria. CONCLUSIONS: We demonstrate for the first time the existence of antibodies to PspA in human sera in health and disease. The findings in ill children suggest that antibodies to PspA might play a role in protection against pneumococcal disease. PMID- 10694001 TI - Prophylaxis for respiratory syncytial virus with respiratory syncytial virus immunoglobulin intravenous among preterm infants of thirty-two weeks gestation and less: reduction in incidence, severity of illness and cost. AB - OBJECTIVE: To determine the impact of respiratory syncytial virus (RSV) prophylaxis among preterm infants of < or =32 weeks gestation by comparing the severity of illness and cost of RSV-related care during the two winter seasons before (1994 to 1995, 1995 to 1996) with the two seasons after initiation of prophylaxis (1996 to 1997, 1997 to 1998). METHODS: Preterm infants of < or =32 weeks gestation at risk for hospitalization with RSV infection were identified retrospectively from the infants hospitalized in our neonatal units. Infants were included if they (1) were born 6 months before or during four winter seasons (1994 to 1998), (2) were discharged from the neonatal unit and (3) had remained in the university outpatient clinic system during at least the first winter of life. Preterm infants of < or =32 weeks gestation hospitalized with RSV were identified from our RSV database (which includes cost of hospitalization, duration of hospital stay, pediatric intensive care unit stay and intubation). Infants receiving prophylaxis were identified prospectively. RESULTS: The incidence of hospitalization with RSV was significantly lower among the cohort of infants born after initiation of prophylaxis: 8.7% (17 of 195) vs. 22% (35 of 159), P = 0.00049 by two tailed Fisher's exact test. Among the cohort of infants born after initiation of prophylaxis (n = 195), 100 infants received prophylaxis. The gestational and chronologic ages of the prophylaxis-treated infants were significantly lower than those of the non-prophylaxis-treated infants (n = 95). The prophylaxis-treated infants also were more likely to have bronchopulmonary dysplasia. Only 1 (1%) of the prophylaxis-treated infants required hospitalization for RSV. Comparison of the cohort of infants born before initiation of prophylaxis to the cohort born after initiation of prophylaxis (includes prophylaxis-treated and non-prophylaxis-treated infants) revealed a significant reduction in severity of illness and cost. The length of stay in the cohort born before initiation of prophylaxis was reduced 83.8%: 373.6 days per 100 infants at risk vs. 60.5 (P = 0.00055). The length of stay in the pediatric intensive care unit was reduced 92.7%: 218.2 days per 100 infants at risk vs. 15.9 (P = 0.00029). The duration of intubation was reduced 95.6%: 187.4 days per 100 infants at risk vs. 8.2 (P = 0.00024). The dollars spent for RSV-related care (hospitalizations and prophylaxis) per 100 infants at risk for RSV was reduced 65% in the cohort of infants born after prophylaxis: $670,590 per 100 infants at risk vs. $234,596 (P = 0.00056). This reduction remained significant (64.9%) if the cost of ribavirin (drug and administration fees) was excluded from the cost of hospitalization. CONCLUSIONS: These data reveal that RSV prophylaxis significantly reduced the incidence of RSV hospitalizations and severity of illness as well as the cost of RSV-related care among these infants. PMID- 10694003 TI - Hyperbaric oxygen. PMID- 10694002 TI - Safety and immunogenicity of three doses of a Neisseria meningitidis A + C diphtheria conjugate vaccine in infants from Niger. AB - BACKGROUND: High rates of endemic disease and recurrent epidemics of serogroup A and C meningococcal meningitis continue to occur in sub-Saharan Africa. A meningococcal A + C polysaccharide diphtheria-toxoid-conjugated vaccine may address this issue. METHODS: In Niger three doses of a bivalent meningococcal A + C diphtheria-toxoid-conjugated vaccine (MenD), containing 1, 4 or 16 microg of each polysaccharide per dose, administered at 6, 10 and 14 weeks of age, were compared with Haemophilus influenzae type b-tetanus toxoid-conjugated (PRP-T) vaccine given with the same schedule or with a meningococcal A + C polysaccharide vaccine (MenPS) given at 10 and 14 weeks of age. One blood sample was taken at the time of enrollment (6 weeks of age) and another was taken 4 weeks after the primary series. RESULTS: All doses of MenD were well-tolerated. After the primary series a higher proportion of infants had detectable serum bactericidal activity against serogroup A for each dose of MenD (from 94% to 100%) than for MenPS (31%) or H. influenzae type b-tetanus toxoid-conjugated vaccine (18.9%); P < or = 0.05. Significant differences were also observed for serogroup C MenD 4 microg or MenD 16 microg (100%) vs. MenPS (69.7%) or Haemophilus influenzae type b-tetanus toxoid-conjugated vaccine (24.3%); P < or = 0.05. When MenPS vaccine was given to 11-month-old children, the immune response measured by both enzyme-linked immunosorbent assay and serum bactericidal assay was greater in those previously immunized with MenD than in those immunized with MenPS vaccine. CONCLUSION: MenD was safe among infants in Niger, and immunization led to significantly greater functional antibody activity than with MenPS. The 4-microg dose of MenD for both the A and C serogroups has been selected for further studies. PMID- 10694004 TI - The relationship between infection with group A beta hemolytic streptococci and the development of psoriasis. PMID- 10694005 TI - Cokeromyces recurvatus as a human pathogenic fungus: case report and critical review of the published literature. PMID- 10694006 TI - Endocarditis caused by Corynebacterium diphtheriae: case report and review of the literature. PMID- 10694007 TI - Respiratory coronavirus infections in children younger than two years of age. PMID- 10694008 TI - Neonatal septicemia caused by Vibrio cholerae O:139. PMID- 10694009 TI - Mefloquine-induced psychosis. PMID- 10694010 TI - Donovanosis causing cervical lymphadenopathy in a five-month-old boy. PMID- 10694011 TI - Herpes zoster in healthy children immunized with varicella vaccine. PMID- 10694012 TI - Severe ehrlichiosis in an adolescent taking trimethoprim-sulfamethoxazole. PMID- 10694013 TI - Severe rhabdomyolysis, hyperthermia and shock after amphotericin B colloidal dispersion in an allogeneic bone marrow transplant recipient. PMID- 10694014 TI - Human monocytic ehrlichiosis in a child with leukemia. PMID- 10694015 TI - Eleven-month-old with recurrent bacterial and aseptic meningitis. PMID- 10694016 TI - Serum alpha-tocopherol and beta-carotene levels are not associated with rheumatic fever in Bangladeshi children. PMID- 10694017 TI - Sibling history of recurrent acute otitis media correlates with low IgG2 anti pneumococcal polysaccharide antibody levels. PMID- 10694018 TI - Whither penicillin? A response to the question. PMID- 10694019 TI - Rubella immunity in young women. PMID- 10694020 TI - Operative repair of the fixed hammertoe deformity. AB - Sixty-three patients (118 toes) were evaluated at an average 61 month follow-up following PIP resection arthroplasty for a fixed hammertoe deformity. The deformity involved the second toe in 35%, the third toe in 21%, the fourth toe in 24%, and the fifth toe in 20%. The involved toe averaged 2 mm. greater length than the adjacent toes and was longer in 49/94 (52%). Seventy-eight percent of patients complained of pain preoperatively due to the hammertoe deformity and 49% complained of callus formation. Following a resection arthroplasty technique with intramedullary Kirschner wire fixation, fusion of the PIP joint occurred in 81% of toes. A fibrous union resulted in the remaining 19% of cases. Patients rated subjective alignment as acceptable in 86% of cases and radiographic alignment was rated as good in 79%. Malalignment and numbness were the major factors associated with an unsuccessful result. Pain was relieved in 92%of patients and subjective satisfaction was noted by 84% of patients. Minor complications occurred in 5%. The average postoperative AOFAS score was 83 points. Resection arthroplasty of the proximal interphalangeal joint with intramedullary Kirschner wire fixation as a technique for correction of a fixed hammertoe deformity is a reliable technique that consistently gives a high level of satisfactory results. PMID- 10694021 TI - Sinus tarsi approach with trans-articular fixation for displaced intra-articular fractures of the calcaneus. AB - The charts and radiographs of 99 patients with 106 intraarticular fractures of the calcaneus were retrospectively reviewed. There were 75 men and 24 women. The average age was forty-two (range, 17 to 81). Fifty-seven of the fractures were left and 49 were right. The mechanism of injury was a fall from a height in 69 patients and motor vehicle accident in 30 patients. According to Sanders classification, seventy-one cases (67%) had type II fractures, 25 cases (23.6%) had type III, and ten cases (9.4%) had type IV. All the patients had operative management through a limited sinus tarsi approach with minimal fixation of the fracture with one or several pins. One of the pins was usually applied from the talus to the calcaneus through the fracture after reduction of the posterior facet. Nine cases (8.5%) developed postoperative infection, four cases (3.8%) had superficial wound infection, four cases (3.8%) had pin tract infection and one case (0.9%) had osteomylitis. Our follow-up at an average of 29 months (range, 12 to 84 months) showed that the American Orthopedic Foot and Ankle Society, Ankle Hindfoot Score for the all group was 77.6 (range, 31-91). Forty-one fractures (38.8%) were graded excellent, 39 fractures (36.7%) good, 14 fractures (13.2%) fair, and 12 fractures (11.3%) were failures. Although radiological degenerative changes in the subtalar joint were seen in 41 cases (38.7%), only six cases (5.6%) required subsequent subtalar fusion. The authors conclude that the operative method used in the current study which followed the principle of minimal soft tissue damage and minimal internal fixation may be a good option for management of calcaneus fractures. PMID- 10694022 TI - Internal architecture of the calcaneus: implications for calcaneus fractures. AB - STUDY DESIGN: Fourteen cadaveri specimens were sectioned to analyze the internal architecture of the human calcaneus. We described the arrangement and orientation of trabecular patterns within the calcaneus and made multiple measurements of its cortical thickness. OBJECTIVE: To characterize the internal architecture of the calcaneus and correlate these findings with well-described patterns of calcaneus fracture in order to better understand the fracture mechanics of this common fracture. METHODS: Fourteen dry, frozen, human calcanei were sectioned using a saw. In each the coronal, sagittal and axial planes, we sectioned separate specimens into slices of 0.5 mm thickness. High-resolution radiographic images were taken of the sectioned specimens. The internal trabecular arrays were described and measurements of cortical thickness were recorded. The correlation between these findings and the known pattern of calcaneal fractures was analyzed. RESULTS: A dominant trabecular pattern running antero-posteriorly along the long axis of the calcaneus was observed. In the posterior tuberosity the trabeculae were arranged parallel to the posterior border. There was an area of sparse or absent mineralization in the anterior part of the calcaneus corresponding to the "neutral triangle" described by Wood and Harty 10, 23. The thickest sites of the calcaneal cortex were the lower pole of the posterior tuberosity, the upper surface at the angle of Gissane, and the lateral surface below the anterior portion of the posterior facet. CONCLUSION: The trabecular architecture of the calcaneus is created by applied stress in concordance with Wolff's law. The weakest plane of resistance to stress is parallel to these organized trabeculae or through areas lacking trabeculae. This study demonstrates that the primary and secondary fracture lines commonly encountered in calcaneus fractures correlates with the internal architectural map of the calcaneal trabecular patterns. PMID- 10694023 TI - Treatment strategies in osteochondral defects of the talar dome: a systematic review. AB - The aim of this study was to investigate the results of different treatment strategies for osteochondral defects (OCD) of the talus. Electronic databases from 1966 to July 1998 were systematically screened. Based on our inclusion criteria 32 studies describing the results of treatment strategies for OCD of the talus were included. No randomized clinical trials (RCT's) were identified. Fourteen studies described the results of excision alone, 11 the results of (EC), 14 the results of (ECD), 1 the results of cancellous bone grafting after EC, 1 the results of osteochondral transplantation and 3 the results of fixation. The average success rate of non-operative treatment (NT) was 45%. Comparison of different surgical procedures shows that the average highest success rate was reached by excision, curettage and drilling (ECD) (85%) followed by excision and curettage (EC) (78%) and excision alone (38%). Based on this systematic review we conclude that NT and excision alone are not to be recommended in treating talar OCD. Both EC and ECD have been shown to lead to a high percentage good/excellent results. However, due to great diversity in the articles and variability in treatment results, no definitive conclusions can be drawn. Further prospective randomized controlled trials are required to compare the outcome of these two surgical strategies for OCD of the talus. PMID- 10694024 TI - Follow-up study of MRI for osteochondral lesion of the talus. AB - Characteristic MRI findings of osteochondral lesions of the talus have been reported. We examined how they change before and after treatment and discussed their significance. Twenty two ankles in 21 patients had MRI examination before and after treatment of the talar lesion. We evaluated the changes in the low intensity areas in T1-weighted image and the signal rims behind osteochondral fragment in T2-weighted image which have been reported as characteristic findings. Clinical symptoms were improved postoperatively in all subjects. The low intensity areas in T1-weighted image seen before the surgical treatment tended to decrease in size postoperatively. The low intensity area in T1-weighted image was reduced in 15 of the 22 ankles (68.2%). Low signal rim in T2-weighted image was seen in three cases before the treatment. All disappeared completely after arthroscopic drilling. Similarly, high signal rim in T2-weighted image seen in 13 cases before the treatment disappeared in 10 postoperatively. These findings were considered indicative that surgical treatments reduced abnormal stress of the underlying bone element due to unstable osteochondral fragment, leading to reduction of the low intensity area. The disappearance of signal rims in T2-weighted images was considered to indicate obliteration of the interface between the osteochondral fragment and the talar bed with bone union. We believe that MRI of the osteochondral lesion of the talus will be useful for postoperative evaluation allowing assessment of the need for further treatment. The decreasing size of low intensity areas in T1-weighted images and disappearance of signal rims behind the osteochondral fragment in T2-weighted images suggested healing of the osteochondral lesions. PMID- 10694025 TI - Transient osteoporosis of the talus. AB - Transient osteoporosis is a clinical syndrome of unknown etiology characterized by the acute onset of pain gradually worsening over several weeks to months. Radiographic changes occur, but laboratory studies are generally unremarkable. Transient osteoporosis of the talus appears to have a similar clinical appearance, radiographic findings, and successful response to conservative management as transient osteoporosis found elsewhere in the body and can be treated similarly. Awareness of this syndrome is important to avoid confusing it with a variety of other disorders of the talus that may have similar clinical presentations. PMID- 10694026 TI - Ankle biomechanics during impact landings on uneven surfaces. AB - Inversion sprains of the lateral ligaments of the ankle are one of the most common of all sporting injuries. While the strains in the anterior talofibular (ATFL) and calcaneofibular (CFL) ligaments have been measured in quasi-static conditions, the dynamic strains during an actual traumatic event have not been determined. The present investigation determined the strains and strain rates in the ATFL and CFL during an in vitro inversion sprain. The ATFL tended to have higher strain and strain rate values than the CFL, which may explain why it is more often injured than the CFL. PMID- 10694027 TI - Periosteal chondroma of the cuboid presenting in a 7-year-old-boy. AB - This case report discusses the finding of a periosteal (juxtacortical) chondroma of the cuboid in a 7-year-old male. While this lesion is well recognized in the tubular bones of adults, this case is unusual due to the child's age, the site of the lesion (cuboid), and the difficulty in establishing the diagnosis due to the cellular atypia. The child was treated with marginal resection and curettage with no evidence of local recurrence on follow-up. A review of the literature is also included in the discussion. PMID- 10694028 TI - Isolated medial cuneiform fracture: review of the literature and report of two cases. AB - The authors present two unusual cases of isolated medial cuneiform fracture. Both fractures were difficult to see on plain films and therefore diagnosed with ancillary tests (computed tomography and magnetic resonance imaging). Treatment was nonweightbearing cast immobilization, in which both patients healed within twelve weeks of treatment without complication and returned to full work related activities. PMID- 10694029 TI - Modification of the Chrisman-Snook technique. PMID- 10694030 TI - Intermediate to long term follow-up of medial approach dorsal cheilectomy for hallux rigidus. PMID- 10694031 TI - Dorsiflexion metatarsal osteotomy for treatment of recalcitrant diabetic neuropathic ulcers. PMID- 10694032 TI - Dorsiflexion metatarsal osteotomy for treatment of recalcitrant diabetic neuropathic ulcers. PMID- 10694033 TI - External rotation-lateral view of the ankle in the assessment of the posterior malleolus. PMID- 10694034 TI - Superoxide and hydroxyl radical generation, and superoxide dismutase in PSII membrane fragments from wheat. AB - Illumination of photosystem II (PSII) membrane fragments of wheat (Triticum durum Desf. cv. Adamello) gave rise to both O2*- and *OH radicals adducts of the novel spin trap 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO). With time, *OH became predominant displaying the conversion of O2*- into *OH. An intrinsic activity of superoxide dismutase (SOD) was found in PSII. Photoreduction of nitroblue tetrazolium (NBT) by PSII membrane fragments was induced by the addition of sodium azide and hydrogen peroxide. Western blotting of PSII proteins showed that a 29 kDa protein was recognised by an antibody against chloroplastic Fe-SOD from water lily. An increased formation rate of O2*- was observed in damaged PSII where the SOD activity decreased following a treatment with a free radical-generating system. Damage in PSII consisted also in a decrease in chlorophyll and in carotenoids as well as in a change in the lipid : chlorophyll : protein ratio. Upon treatment a decrease in the unsaturation of PSII membrane fragments was also monitored together with a degradation towards more saturated molecular species of monogalactosyldiacylglycerol. PMID- 10694035 TI - Differential response of antioxidative enzymes of chloroplasts and mitochondria to long-term NaCl stress of pea plants. AB - In this work the activity of superoxide dismutase (SOD) and the enzymes of the ascorbate-glutathione (ASC-GSH) cycle were investigated in chloroplasts and mitochondria from leaves of Pisum sativum L. cv. Puget after 15 days treatment with 0-130 mM NaCl. The main chloroplastic SOD activity was due to CuZn-SOD II, which was increased significantly (about 1.7-fold) by NaCl, although during severe NaCl stress (110-130 mM) chloroplastic Fe-SOD exhibited a stronger enhancement in its activity (about 3.5-fold). A sudden induction in chloroplastic APX, DHAR and GR was also caused by NaCl (70-110 mM), but not by the highest salt concentration (130 mM), at which GR and DHAR activities were similar to the control values and APX decreased. In addition, the H2O2 concentration and lipid peroxidation of membranes increased significantly, 3.5- and 7-fold, respectively, in chloroplasts under severe NaCl stress. In purified mitochondria DHAR and GR were significantly induced only at 90 and 130 mM NaCl, respectively, although DHAR activity was below control values in the highest NaCl concentrations. APX and MDHAR activities started their response to salt in mild NaCl conditions (70 mM) and increased significantly with the severity of the stress. Mn-SOD was induced only under severe NaCl concentrations. The mitochondrial H2O2 and lipid peroxidation were increased at the highest NaCl concentration although to a lesser extent (about 2-2.5-fold) than in chloroplasts, whereas the increase in carbonyl protein contents was higher in mitochondria. The results suggest that the degree of enhanced tolerance to NaCl seems to require the induction of specific isoforms, depending on the different organelles. PMID- 10694036 TI - Ozone detoxification in the mesophyll cell wall during a simulated oxidative burst. AB - With the aim to assess the effect of possible O2*- generation during an oxidative burst on O3 reduction in mesophyll cell walls due to the reaction O2*- + O3 --> O3*- + O2 and subsequent formation of *OH, 03 flow through this sequence was compared with O2*- flow through the competitive sequences where H2O2 is formed. The two-electron reduction of O3 via the direct reaction with ascorbate was also considered. The calculations were exemplified in an experiment where Phaseolus vulgaris L. leaves were exposed to 530 nl O3 l(-1) in air for 3.5 h. During the exposure, H2O2 was assumed to be generated at peak rates observed in pathogen elicited cell suspensions. O3 reduction through reaction with O2*- was 25-44% of O3 detoxified in the direct reaction with ascorbate. More than 99% of O2*- was reduced to H2O2 via spontaneous disproportionation and reduction with ascorbate, the disproportionation prevailing at pH 5 and reduction at the expense of ascorbate at pH 7. H2O2 was estimated to be channelled mostly to the peroxidase catalysed scavenging reaction. Calculated steady state H2O2 concentrations were 40-80 microM. It is concluded that generation of H2O2 at the postulated rate was too high and that the generation of O2*- during an oxidative burst is ineffective in reducing O3 through the network of reactive oxygen species. Superoxide dismutase induction in the cell wall under O3 is discussed. PMID- 10694037 TI - Cadmium toxicity and oxidative metabolism of pea leaf peroxisomes. AB - The effect of growing pea plants with 50 microM CdCl2 on the activated oxygen metabolism was studied at subcellular level in peroxisomes isolated from pea leaves. Cadmium treatment produced proliferation of peroxisomes as well as an increase in the content of H2O2 in peroxisomes from pea leaves, but in peroxisomal membranes no significant effect on the NADH-dependent O2*- production was observed. The rate of lipid peroxidation of membranes was slightly decreased in peroxisomes from Cd-treated plants. This could be due to the Cd-induced increase in the activity of some antioxidative enzymes involved in H2O2 removal, mainly ascorbate peroxidase and glutathione reductase, as well as the NADP dependent dehydrogenases present in these organelles. The activity of xanthine oxidase did not experiment changes by Cd treatment and this suggests that O2*- production in the peroxisomal matrix is not involved in Cd toxicity. This was supported by the absence of changes in plants treated with Cd in the Mn-SOD activity, responsible for O2*- removal in the peroxisomal matrix. Results obtained indicate that toxic Cd levels induce imbalances in the activated oxygen metabolism of pea leaf peroxisomes, but its main effect is an enhancement of the H2O2 concentration of these organelles. Peroxisomes respond to Cd toxicity by increasing the activity of antioxidative enzymes involved in the ascorbate glutathione cycle and the NADP-dependent dehydrogenases located in these organelles. PMID- 10694038 TI - Cd-induced oxidative burst in tobacco BY2 cells: time course, subcellular location and antioxidant response. AB - The relation between Cd and oxidative stress in BY2 cell cultures of tobacco was studied. In response to 5 mM Cd, a rapid generation of H2O2 has been detected in tobacco cell cultures by the oxidative quenching of the fluorescent reporter dye pyranine. This oxidative burst reached the maximum production of H2O2 after 10 min of treatment with Cd. This response could be considered as short term hypersensitive response previous to the oxidative stress caused by the metal at the cell plasma membrane. The observed antioxidant enzymatic response to the oxidative burst was preceded by an increased peroxidation of lipids with a significant increase in the activities of superoxide dismutase and ascorbate peroxidase. The results presented in this study point out to the plasma membrane as the primary target for the short term production of activated oxygen species in response to Cd in BY2 tobacco cells followed by a coordinated activation of the antioxidant enzymatic system. PMID- 10694039 TI - The chemical behaviour of heavy metals plays a prominent role in the induction of oxidative stress. AB - It is often described that different environmental stress factors stimulate the production of reactive oxygen species and increase the activity of several enzymes quenching these radicals. The ascorbate-glutathione pathway is also involved in plant defence against oxidative stress. Therefore the effects of 2 metals (Cu, Zn) with different chemical behaviour were investigated on the enzymes of this pathway in the primary leaves of bean seedlings grown on hydroponics and supplied with a 50 microM concentration of both metals. The results obtained demonstrate that the capacities of the enzymes involved in the ascorbate-glutathione pathway increase after metal application, indicating that they induce oxidative stress indeed. However striking differences in the relative induction time of these enzymes suggest that the chemical behaviour of the metals applied, plays an important role in the induction of oxidative stress as well as in the defence mechanism against it. PMID- 10694040 TI - Oxidative stress induced by copper in sunflower plants. AB - The toxic effect of copper was evaluated in sunflower plants by means of root elongation, lipid peroxidation and antioxidative enzyme activities. Plants were grown under controlled environmental conditions in hydroponic culture for two weeks and exposed to different copper concentrations for an additional week. Determinations were performed in both roots and leaves. From the results of the root elongation test, a copper concentration of 10 microM was found to be toxic for sunflower plants causing lipid peroxidation and, thus, oxidative stress. Antioxidative enzymes (peroxidase and superoxide dismutase) responded at lower, nontoxic copper concentrations. PMID- 10694041 TI - Inhibition of catalase by sulfite and oxidation of sulfite by H2O2 cooperating with ascorbic acid. AB - An oxidative detoxification of sulfite, which originates from sulfur dioxide taken up into a leaf, has not yet been fully understood. In this study, we discuss that redox reactions between sulfite and H2O2 have an important role for the detoxification of sulfite. Sulfite was oxidized by H2O2 and during the redox reaction, oxygen consumption was observed. The oxygen consumption was partially inhibited by superoxide dismutase, indicating that O2- is generated during the redox reaction. Oxidation of sulfite by H2O2 was also observed in the presence of ascorbic acid, and during the oxidation, no significant oxidation of ascorbic acid and no consumption of oxygen were observed. Sulfite inhibited catalase of cell-free extracts of spinach, pea and broad bean leaves. These results suggest that when leaves are fumigated with SO2 in the light, catalase is inactivated resulting in the accumulation of H2O2 in leaves, which can oxidize sulfite without generating active oxygen species like O2- as long as ascorbate is present in leaves. PMID- 10694042 TI - Antioxidative defense and photoprotection in Pinus halepensis induced by Mediterranean conditions and ozone exposure. AB - Pigment levels and antioxidative stress-related metabolites were determined over a period of two years in Pinus halepensis to characterize the response of some plant protective processes to both environmental stresses and 03 exposure. Two year-old-Aleppo pine seedlings were grown in open-top chambers under optimal nutrient and water conditions and exposed to different 03 levels: charcoal filtered air, non-filtered air and nonfiltered air plus 40 ppb O3 10 h/day. In summer an activation of photoprotective systems was observed since a decrease in chlorophyll levels, an increase in the carotenoid/chlorophyll ratio and SOD activation were recorded. Interestingly, the SOD activity in Pinus halepensis was highly related to low-molecular weight compounds and this relationship increased with needle age. Ozone exposure induced alterations in the activity of some antioxidant enzymes along with reductions in pigment concentrations and an activation of the xanthophyll cycle. PMID- 10694043 TI - Responses of the xanthophyll cycle pool and ascorbate-glutathione cycle to ozone stress in two tobacco cultivars. AB - Plants of Nicotiana tabacum (O3-tolerant cv Bel-B and O3-sensitive cv Bel-W3) were exposed to 150 ppb of ozone for 5 h; the fumigation produced visual injury in mature leaves, particularly in Bel-W3. After O3-treatment the pigments of the xanthophyll cycle pool decreased in both cvs, with a strong reduction in violaxanthin content, while antheraxanthin and zeaxanthin increased slightly. Under these conditions the content of leaf abscisic acid (ABA) markedly increased, particularly in O3-sensitive cv, indicating that the violaxanthin may have been partially converted into ABA. The control plants of Bel-B showed an ascorbic acid content four times greater than Bel-W3 and the ozone treatment did not produce significant differences in the ascorbic acid content and in the redox state. The two tobacco cvs were found to have similar total glutathione content, however the redox state was lower in O3-sensitive cv and decreased after ozone exposure. Ozone fumigation caused an increase in oxidized glutathione, particularly in Bel-W3, associated with a reduced glutathione reductase (GR) activity and a reduced GR protein content. PMID- 10694044 TI - Health effects and bioavailability of dietary flavonols. AB - Flavonoids are polyphenolic compounds that are ubiquitously present in foods of plant origin. Flavonoids are categorised into flavonols, flavones, catechins, flavanones, anthocyanidins, and isoflavonoids. They may have beneficial health effects because of their antioxidant properties and their inhibitory role in various stages of tumour development in animal studies. It is estimated that the human intake of all flavonoids is a few hundreds of milligram per day. Flavonoids present in foods used to be considered non-absorbable because they are bound to sugars as beta-glycosides. However, we found that human absorption of the quercetin glycosides from onions (52%) is far better than that of the pure aglycone (24%). The sugar moiety is an important determinant of their absorption and bioavailability. Flavonol glycosides might contribute to the antioxidant defences of blood. The average intake of the flavonols quercetin, myricetin and kaempferol and the flavones luteolin and apigenin in the Netherlands was 23 mg/day. The intake of these flavonols and flavones was inversely associated with subsequent coronary heart disease in some but not all prospective epidemiological studies. A protective effect of flavonols on cancer was found in one prospective study; two others showed no association. Thus the epidemiological evidence does not yet allow a decision on the involvement of flavonols in the etiology of either cardiovascular diseases or cancer. PMID- 10694045 TI - Inhibition of biochemical model reactions for inflammatory processes by plant extracts: a review on recent developments. AB - All processes of oxygen activation include very reactive intermediates. Therefore, aerobic cells must cope with- and to some extent also adapt to- oxidative stress provoked for example by infections or intoxications, where these reactive intermediates accumulate. Dependent on the strength of these impact, several symptoms indicate the deviation from normal, steady-state-metabolism. Intrinsic radical scavenging processes or compounds administered with food thus have to warrant metabolic control within certain limits. Antioxidants which in many cases are free radical scavengers or quenchers of activated states comprise a wealth of classes of organic molecules including phenolics, probably as the most prominent ones. In this communication mechanisms of protection from oxidative damage are discussed. Furthermore, examples of antioxidative functions of a few important natural products in certain diseases are reported. PMID- 10694046 TI - Methods to measure the antioxidant activity in plant material. A comparative discussion. AB - Methods to determine total antioxidant activity (TAA) are generally based on the inhibition of certain reactions by the presence of antioxidants. The most widely used methods are those that involve the generation of radical compounds, and it is the presence of antioxidants that determines the disappearance of these radicals. Strategies for measuring TAA are discussed, particularly the different methodological and instrumental approaches used. In addition, our own methods are presented in order to facilitate and speed up the manipulation of biological material. The values of TAA by different compounds are presented and compared. PMID- 10694047 TI - Photooxidative bleaching of chlorophyllin. AB - Chlorophyllin, a water-soluble, copper-containing porphyrin, can be bleached rapidly in the light or slowly in the dark in a reaction which is oxygen dependent. Both the photo and the dark bleaching reactions are temperature dependent. Cyclic voltammetry measurements show that the copper in the bleached and nonbleached state remains in the +2 redox state and could be readily reduced. This would imply that there is no net oxidative change to the copper during the bleaching process. FT-IR absorption spectroscopy showed vibrations characteristic of a vinyl functionality disappeared upon bleaching. Aqueous solutions of chlorophyllin were not dialyzable through dialysis tubing of molecular weight cut off, 6000-8000 molecular weight, indicative of an aggregate chlorophyllin micelle. Analysis of products by high performance liquid chromatography showed that the chlorophyllin mixture was more complex than originally anticipated and that two components were lost from the mixture upon photobleaching. One compound that is preferentially lost upon photobleaching has been identified by mass spectral analysis as Cu(II) chlorin e6. PMID- 10694048 TI - Response of abietane diterpenes to stress in Rosmarinus officinalis L.: new insights into the function of diterpenes in plants. AB - Abietane diterpenes were measured in field-grown rosemary (Rosmarinus officinalis L.) plants throughout the year. Carnosic acid and carnosol, which were present in high amounts (5 and 1 mg/gDW respectively) in rosemary leaves, decreased by ca. 50% during the Mediterranean summer in response to low precipitation, high radiation and high temperature. In contrast, the highly oxidised diterpenes rosmanol, isorosmanol and dimethyl isorosmanol, which are formed from carnosic acid by enzymatic dehydrogenation and the action of activated oxygen, increased in response to such environmental constraints. Collectively, these data support the contention that abietane diterpenes from R. officinalis function as antioxidants that protect biological membranes from oxidative stress. This is especially important during the Mediterranean summer when there is low precipitation, high light and high temperature. PMID- 10694049 TI - Antioxidants and protective pigments of Pinus ponderosa needles at gradients of natural stresses and ozone in the San Bernardino Mountains in California. AB - At the San Bernardino Mountains, California, a well documented gradient of ozone pollution overlays a natural stress gradient from mesic to dry and from lower elevation to higher elevation sites. In contrast to gradient studies in European regions, the highest ambient ozone levels are observed at low elevation and more mesic locations. In the present study, antioxidative and photoprotective systems in Pinus ponderosa needles were investigated at three plots--DW (1725 m, high ozone impact, mesic site), SW (1200 m, clean air, xeric site) and CO (above 2000 m, clean air and xeric site). Needles from the CO site contained significantly more total GSH (500 vs 300 nmol g(-1) dw in c needles), less alpha-carotene (6-10 vs 14-19 microg mg(-1) total chlorophyll) and chlorophyll (1.7-2 vs 2.5-2.6 mg g( 1) dw in c + 1 needles) than those at the DW site. Furthermore, their xanthophyll cycle pool was in a more de-epoxidized state at midday (up to 60% in c needles), and the carotenoid/chlorophylls ratios were generally higher. These patterns correspond to those observed at higher elevation plots in the Alps. On the other hand, needles from the high ozone site (DW) had a higher proportion of GSSG, indicating the onset of biochemical injury to needles. Needles from the SW site had intermediate proportions of GSSG. The results show the potential of environmental stressors to induce antioxidative and photoprotective responses in the absence of elevated ozone concentrations, but support the oxidative effects of ozone injury to ponderosa pine. PMID- 10694050 TI - Effects of an oxidizing agent (hydrogen peroxide) on the glutathione system in epidermal cells of Allium cepa L. investigated by histochemical staining. AB - The glutathione system in epidermal cells of Allium cepa L. was measured by a quantitative image analysis method, using histochemical staining by monochlorobimane. Blockage of thiol groups with N-ethylmaleimide decreased fluorescence to a small rest fluorescence of maximum 5%. Pre-treatment with dithiothreitol increased fluorescence yield in all cells compared to the monochlorobimane treatment alone. These results correspond to the blockage or the reduction of the total pool of glutathione in the cells. After treatment with N ethylmaleimide and subsequent incubation with dithiothreitol followed by labelling with bimane a low fluorescence yield was observed which correspond to the proportion of GSSG. Specimens that were incubated in H2O2 prior to the treatments described above exhibited a decrease in total glutathione and in reduced glutathione and an increase in the proportion of GSSG compared to the control. PMID- 10694051 TI - Vitamin E supplementation increases the stability and the in vivo antioxidant capacity of refined olive oil. AB - Two experiments were carried out to investigate if the supplementation with vitamin E affects refined olive oil response to oxidation regarding the stability of the oil and the protection in vivo against lipid peroxidation in rats after its intake in comparison with other edible oils. In experiment 1, samples of virgin olive oil, refined olive oil, refined olive oil supplemented by us with 200 mg/kg vitamin E, and sunflower oil were collected before and after a 60 min frying process. After frying, refined olive oil supplemented with vitamin E compared with the non-supplemented refined olive oil had a higher concentration of alpha-tocopherol (240.34+/-6.07 mg/kg vs. 131.94+/-8.14 mg/kg), more resistance against oxidation (19.01+/-1.88% vs. 10.6+/-2.08%) and less polar components (4.2+/-0.06% vs. 5.45+/-0.22%). In experiment 2, 24 male Wistar rats, divided into 4 groups, were fed on diets based on the same unfried oils (8% w/w) as in experiment 1, for 4 weeks. Two days prior to the end of the experiment, the rats were intraperitoneally administered with adriamycin (10 mg/kg/ day) to provoke an oxidative stress. The rats fed on refined olive oil plus vitamin E compared to the rats fed on non-supplemented refined olive oil had lower hydroperoxides concentrations (26.8+/-2.6 nmol/mg vs. 35.6+/-2.49 nmol/mg) higher coenzyme Q levels (128.1+/-11.97 pmol/mg vs. 81.25+/-9.25 pmol/mg) and higher alpha-tocopherol values (1.23+/-0.04 mmol/mg vs. 0.93+/-0.06 mmol/mg) in microsomes of liver. In conclusion, the supplementation of refined olive oil with 200 mg/kg of vitamin E increases the stability of this oil under pro-oxidant conditions, and its intake decreases the oxidative damage generated by adriamycin in rats. PMID- 10694052 TI - Recent advances in understanding the origin of the apoplastic oxidative burst in plant cells. AB - The origin of the oxidative burst during plant-pathogen interactions remains controversial. A number of possibilities have been identified, which involve the protoplast, plasmalemma or apoplast. The apoplastic production of H2O2 requires three components, an extracellular peroxidase, ion fluxes leading to extracellular alkalinisation and release of a substrate. Fatty acids are the major compounds that appear in the apoplast following elicitation, which can activate H2O2 production by peroxidases in vitro. However, the reaction with peroxidases appears to be novel and is uncharacterised at present. The apoplastic mechanism also cannot be readily distinguished from the operation of a plasma membrane NADPH oxidase system by the use of the inhibitors diphenylene iodonium and N,N diethyl-dithiocarbamate since it is also inhibited by these. These inhibitors have often in the past been used to define the involvement of the latter in the oxidative burst. In common with the NADPH oxidase system, the peroxidase responsible has been cloned but unlike the NADPH oxidase it has been shown to function in vitro to generate H2O2. In vivo studies of the oxidative burst have shown that the alkalinisation is essential and the underlying ion fluxes may be regulated by cAMP. Calcium fluxes are also essential. Although the oxidative activity of peroxidase requires calcium the fluxes have obvious other function. These may include activation of release of substrate and through the activation of a CDPK, regulation of enzymes involved in phytoalexin and cell wall phenolic production such as PAL. PMID- 10694053 TI - Some properties of the H2O2/O2- generating system from the lignifying xylem of Zinnia elegans. AB - The properties of the enzymatic system responsible for generating H2O2/O2- in the lignifying xylem of Zinnia elegans were studied using the starch/KI method for monitoring H2O2 production and the nitroblue tetrazolium method for monitoring superoxide anion production. The results showed that H2O2/O2- production by lignifying xylem tissues was insensitive to inhibitors of peroxidase and poly(di)amine oxidases. However, H2O2/O2 production in the xylem of Z. elegans was sensitive to the inhibitors of phagocytic plasma membrane NADPH oxidase, pyridine, imidazole, quinacrine and diphenylene iodonium. The sensitivity of H2O2/O2- production to the respective inhibitors of calmodulin (R-24571), phospholipase C (neomycin sulfate), and protein kinase (staurosporine), and its reversion by an inhibitor of protein phosphatases (cantharidin); pointed to the analogies existing between the mechanism of H2O2/O2- production in the lignifying xylem of Z. elegans and the oxidative burst observed during the hypersensitive plant cell response. These results suggest the existence of a metabolic cascade involving calmodulin, IP3 and protein phosphorylation in the activation of the enzymatic system responsible for H2O2/O2- production in the lignifying xylem of Z. elegans. PMID- 10694054 TI - Elevation of glutathione level and activation of glutathione-related enzymes affect virus infection in tobacco. AB - The effects of two chemicals, L-2-oxothiazolidine-4-carboxylic acid (OTC) and (S) carvone, were investigated on the development of necrotic symptoms and on the virus concentration in tobacco mosaic virus (TMV)-infected tobacco plants. OTC treatments markedly increased the cellular glutathione (GSH) levels in tobacco leaf discs. In addition, OTC pretreatment considerably decreased both the number of necrotic lesions and the virus content in TMV-infected leaf discs. The monoterpene (S)-carvone increased only slightly the GSH content of leaf tissues and caused lipid peroxidation. (S)-carvone dramatically induced the activity of glutathione S-transferase and to a lesser extent elevated also the activities of ascorbate peroxidase and glutathione reductase. Treatments with (S)-carvone strongly reduced the number and size of necrotic lesions, but did not influence the virus concentration. The results show that increased levels of GSH and activities of GSH-related enzymes by OTC and (S)-carvone reduce necrotization of virus-infected tissues. However, virus multiplication and lesion formation do not necessarily correlate: virus multiplication is suppressed only by substantially elevated GSH contents. PMID- 10694055 TI - Dark stained tissues of the epicarp of encore mandarin: interactions with the production of hydroxyl radicals. AB - The interactions between hydroxyl radical production and the composition of the epicarp cells associated with the dark stained tissues of mature fruits of Encore mandarin were investigated. Phosphatidylinositol content of the cells associated with dark stains was lower in unpitted tissues, whereas the concentration of phosphatidylglycerol, phosphatidylcholine and phosphatidylethanolamine did not vary significantly. In these pitted cells, protein content also showed a 1.59 fold increase. Additionally, the relative proportions of gln, thr, asp, glu and gly increased sharply, while ala and tyr decreased. The polypeptide patterns showed quantitative changes when samples of stained and unstained tissues were compared by SDS-PAGE electrophoresis. Qualitatively, the cells associated with pitted tissues revealed a new polypeptide band with an apparent mol. wt. of 50.4 kDa as well as the disappearance of another one of 10 kDa. Ethylene production in the dark stained tissues was lower than in unpitted tissues and these cells showed significant increases in membrane permeability, hydroxyl radical production, and lipid peroxidation. Furthermore, in these tissues the levels of the carotenoids increased significantly but levels of chlorophyll decreased. It was concluded that in the pitted tissues the significant changes of the membrane composition are closely associated with an increasing acyl lipid peroxidation mediated by hydroxyl radical production. The modulation of this metabolism further indicates an incomplete degradation of fatty acids. The implications of the increasing accumulation of carotenoids in the synthesis of oxy radicals are also discussed. PMID- 10694056 TI - Ascorbate metabolism in potato leaves supplied with exogenous ascorbate. AB - Photosynthesis and leaf ascorbate were measured in potato (Solanum tuberosum L.) plants grown in low light and then transferred to high light. Total foliar ascorbate content in low light-grown plants was 4.72+/-0.42 micromol mg(-1)chl. Over 80% of the ascorbate pool was found in the reduced form irrespective of position on the stem. No statistically-significant light-dependent effects were observed. Leaf discs supplied with [14C]-ascorbate in the dark showed significant ascorbate uptake such that after a 16h incubation over half of the total ascorbate pool in the discs was labelled [14C]-ascorbate. No ascorbate efflux from the leaves occurred during the period of [14C]-ascorbate uptake. The total amount of ascorbate did not increase, however, implying modified ascorbate turnover. The turnover of the [14C]-ascorbate in the leaves occurred at similar rates in both light and darkness. Little degradation of labelled ascorbate was observed, suggesting that uptake of exogenous ascorbate leads to inhibition of de novo ascorbate biosynthesis in potato leaf discs. PMID- 10694057 TI - Preliminary studies of ascorbate metabolism in green and albino regions of variegated leaves of Coleus blumei, Benth. AB - Green and white variegated leaves of Coleus blumei, Benth. were separated into albino and green sections and used to determine the distribution of vitamin C and L-galactose dehydrogenase activity, an enzyme supposed to be involved in ascorbate metabolism, in heterotrophic and autotrophic foliar fractions. Both green and white sections contained vitamin C and activity of L-galactose dehydrogenase. However, in the white parts mainly dehydroascorbate was found, whereas in the green parts the redox state of the ascorbate system varied with light or dark conditions. Characterisation of L-galactose dehydrogenase from illuminated green leaf sections showed increasing activity with increasingly alkaline pH-values and a temperature optimum of 25 degrees C. Since these properties were slightly different than those of L-galactose dehydrogenase activities obtained from albino or darkened green leaf sections, we suggest that the enzyme may be light-modulated. PMID- 10694058 TI - Chlorophyll high-temperature thermoluminescence emission as an indicator of oxidative stress: perturbating effects of oxygen and leaf water content. AB - A high-temperature chlorophyll thermoluminescence emission can be observed in plant leaves, without preillumination, following various oxidative stresses. This emission was recorded from 0 degrees C to 160 degrees C on a leaf sample pressed by a teflon ring on a heating plate. A band at 140 degrees C was observed in senescing tree leaves, a band at 130 degrees C with a shoulder at 75 degrees C in tobacco leaves treated with the fungal elicitor cryptogein. Measurements in pure O2 or N2 atmospheres did not affect the 130/140 degrees C band, although oxygen increased the thermoluminescence emission at higher temperatures and should be consequently eliminated during measurements. Preventing desiccation by covering the sample with a glass window suppressed the 130/140 degrees C band, except for its rising edge at about 80/90 degrees C. This broad 130/140 degrees C band results from the chlorophyll-exciting thermolysis of lipid peroxides, but it disappears when water is kept in the sample up to 100 degrees C, due to a non radiative hydrolysis of the peroxides. Therefore, high-temperature thermoluminescence measurements should be performed on leaf samples allowed to dry under an oxygen-free atmosphere. PMID- 10694059 TI - Does ozone exposure protect from photoinhibition? AB - Tropospheric ozone and high light intensities are two stress factors that often occur simultaneously under natural conditions. Ozone is well known to form oxygen radicals in the apoplastic water and long lasting photoinhibition can cause photooxidative damage also by formation of several species of oxygen radicals. We were interested whether moderate levels of ozone would be able to modulate the response of leaves to photoinhibitory conditions naturally occurring around noon on a bright day. Cuttings of Populus sp. were cultivated in two separate greenhouse-compartments adapted as fumigation chambers. In the two compartments plants were grown in ambient air containing about 20 nmol mol(-1) ozone and in elevated ozone concentrations supplied for 8 h per day. During the midday of bright days Fv/Fm decreased by the same amount in all leaves, indicating photoinhibition. At the same time Fo increased in control leaves more than in ozone-exposed leaves indicating a higher amount of heat-deactivating PSII centres in the latter. This was confirmed by a higher epoxidation state in ozone-exposed leaves during midday of a bright day. The contents of chlorophyll a and chlorophyll b were significantly decreased in ozone-exposed leaves. In older leaves the ratios chlorophyll a : b, and xanthophylls: chlorophyll b were increased indicating an adaptation to higher light stress. From this we conclude that by increasing the amount of heat-deactivating centres ozone seems to protect PSII from photoinhibition. PMID- 10694060 TI - Activated oxygen generation from thylakoids: a novel spin trap. AB - Using a novel phosphorylated spin trap, 5-(diethoxyphosphoryl)-5-methyl-1 pyrroline-N-oxide (DEPMPO), an analogue of the commonly used spin trap 5,5' dimethyl-1-pyrroline N-oxide (DMPO), we have investigated the production of oxygen radical species under illumination of thylakoids from wheat (Triticum durum Desf cv. Ofanto). DEPMPO reacted with superoxide (O2*-) and hydroxyl radical (HO*) forming distinctive spin trap adducts. Spectra of (O2*-) and HO* adducts of DEPMPO were recorded in the presence of xanthine/xanthine oxidase and FeSO4/H2O2, respectively, and computer simulation of spectra was performed. During illumination of thylakoids both O2*- and HO* were detected as well. Transition metals catalysed transformation of O2*- into HO*. The conversion was enhanced by H2O2 and prevented by exogenous superoxide dismutase and catalase. The presence of a thylakoid-bound superoxide dismutase, whose activity was inhibited by H2O2 and diethyldithiocarbamic acid, was responsible for H2O2 production from O2*- and thus for HO* generation. PMID- 10694061 TI - Nitric oxide generation by soybean embryonic axes. Possible effect on mitochondrial function. AB - Nitric oxide (NO) generation and its effect on mitochondrial enzymes were investigated in soybean embryonic axes at the onset of germination. NO was detected in homogenates from soybean embryonic axes by EPR. Enzymatic sources of NO, such as nitrate reductase activity and nitric oxide synthase, assessed as NADPH-diaphorase activity, were measured in homogenates incubated up to 48 h. Both NO content and the activity of the enzymes showed a similar profile as function of the imbibition time, with maximal levels at 15-24h. Total O2 consumption in enriched-mitochondrial fraction was inhibited by NO in a concentration-dependent manner. O2 consumption dependent on cytochrome oxidase activity was more sensitive than alternative oxidase pathway to NO exposure. Half maximal effects of NO at 0.3 and 3.6 microM were measured for cytochrome oxidase and alternative oxidase, respectively. Enriched-mitochondrial fractions from soybean embryonic axes treated with NO (up to 1 microM) showed increased H2O2 production. The data presented suggest that NO could modulate O2 consumption in soybean embryonic axes. This process could affect the pro-oxidant/antioxidant balance and the cellular energy yield in the germinating embryonic axes, and could have a role in soybean germination. PMID- 10694062 TI - Characterisation of a cDNA encoding gamma-glutamylcysteine synthetase in Medicago truncatula. AB - A gamma-ECS cDNA from Medicago truncatula was isolated using an Arabidopsis thaliana cDNA as probe. The analysis of the amino acid sequence deduced from this cDNA revealed 80% identity with the gamma-ECS from A. thaliana and Brassica juncea and suggested a plastidial localisation for the enzyme. Gamma-ECS activity and high level of GSH were detected in the gamma-ECS-deficient E. coli strain expressing a fusion protein containing the M. truncatula gamma-ECS protein. Southern blot analysis suggests that gamma-ECS is encoded by a small multigenic family in M. truncatula and shows that homologous genes are present in two other leguminous plants, Medicago sativa and Pisum sativum. Gamma-ECS gene expression was analysed by Northern blot in seedlings, plantlets and mature plants. PMID- 10694063 TI - Differential gene expressions of rice superoxide dismutase isoforms to oxidative and environmental stresses. AB - Active oxygen species (AOSs) are produced under stress conditions of plant cells. Superoxide dismutase (SOD) catalyzes the first step in the AOS scavenging system. The responses of SOD genes to environmental stresses were analyzed in rice seedlings by the treatments of drought, salinity and chilling. The expressions of abscisic acid (ABA)-inducible genes, Mn-SOD gene (sodA1) and one of the cytosolic Cu/Zn-SOD genes (sodCc2), were strongly induced by the treatment of drought and salinity. While Fe-SOD gene (sodB) and the other cytosolic Cu/Zn-SOD gene (sodCc1) were also induced by ABA. However the mRNA level of sodB was decreased by drought treatment, and sodCc1 gene was not induced by drought and salinity treatments. Plastidic Cu/Zn-SOD gene (sodCp) quickly responded to salinity treatment in the light but not in the dark. In the treatment with hydrogen peroxide, sodCp gene was strongly induced shortly after the treatment. These results suggested that phytohormone and AOSs are associated with the regulation of SOD genes under environmental stresses. PMID- 10694064 TI - Sunflower (Helianthus annuus) variability in antioxidant enzyme defenses. AB - In order to investigate the existence of genetic variability in antioxidant enzyme defenses in sunflower, twelve inbred lines, six cytoplasmic male-sterile and six restorer lines, commonly used in breeding programs have been compared with respect to (a) their levels of constitutive superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), glutathione reductase (GR, EC 1.6.4.2) and guaiacol-dependent peroxidase (GPX, EC 1.11.1.7), and (b) their isoenzyme polymorphism in SOD, CAT, and GPX activities. Constitutive levels of antioxidant enzymes in the 2nd leaf pair of 15-20-day-old sunflower plants showed significant differences between lines. The ranges of variation in enzyme activities of the different lines were equivalent to 34.3% (CAT), 38.2% (SOD), 59.5% (APX), 60.0% (GR), and 62.9% (GPX) of the respective maximal values. Isoenzyme profiles of CAT, GPX and SOD revealed the existence in sunflower of at least three, six and four isoforms of these enzymes, respectively. Further characterization of SOD isoenzymes revealed that no isoenzyme corresponded to a Mn-SOD, the faster moving isoform being a Cu/Zn-SOD and the remainder three Fe-SODs. Among the twelve inbred sunflower lines studied there were ample qualitative, and sometimes quantitative too, differences in isoenzyme dotation of CAT, GPX and Fe-SOD. PMID- 10694065 TI - Purification of catalase from pea leaf peroxisomes: identification of five different isoforms. AB - Catalase activity was analyzed in seven organs of pea (Pisum sativum L.) plants: leaves, seeds, flowers, shoots, whole fruits, pods and roots. Leaves showed the highest activity followed by whole fruits and flowers. Catalase was purified from pea leaf peroxisomes. These organelles were isolated from leaves by differential and sucrose density-gradient centrifugation, and catalase was purified by two steps involving anion exchange and hydrophobic chromatography using a Fast Protein Liquid Chromatography system. Pure catalase had a specific activity of 953 mmol H2O2 min(-1) mg(-1) protein and was purified 1000-fold, with a yield of about 19 microg enzyme per kg of pea leaves. Analysis by SDS-PAGE and immunoblot showed that the pea catalase was composed of subunits of 57 kDa. Ultraviolet and visible absorption spectra of the enzyme showed two absorption maxima at 252 and 400 nm with molar extinction coefficients of 2.14 x 10(6) and 7.56 x 10(6) M(-1) cm(-1), respectively. By isoelectric focusing (pH 5-7), five different isoforms were identified and designated as CAT1-5, with isoelectric points of 6.41, 6.36, 6.16, 6.13 and 6.09, respectively. All the catalase isoforms contained a subunit of 57 kDa. Post-embedment, EM immunogold labelling of catalase showed a uniform distribution of the enzyme inside the matrix and core of pea leaf peroxisomes. PMID- 10694066 TI - Catalase-peroxidases in cyanobacteria--similarities and differences to ascorbate peroxidases. AB - Cyanobacteria (blue-green algae) are oxygenic phototrophic bacteria carrying out plant-type photosynthesis. The only hydrogen peroxide scavenging enzymes in at least two unicellular species have been demonstrated to be bifunctional cytosolic catalase-peroxidases (CatPXs) having considerable homology at the active site with plant ascorbate peroxidases (APXs). In this paper we examined optical and kinetic properties of CatPXs from the cyanobacteria Anacystis nidulans and Synechocystis PCC 6803 and discuss similarities and differences to plant APXs. Both CatPXs and APX showed similar spectra of the ferric enzyme, the redox intermediate Compound I and the cyanide complex, whereas the spectrum of CatPX Compound II had characteristics reminiscent of the spectrum of the native enzyme. Both steady-state and multi-mixing transient-state kinetic studies were performed in order to characterize the kinetic behaviour of CatPXs. Bimolecular rate constants of both formation and reduction of a CatPX Compound I are presented. Because of its intrinsic high catalase activity (which cannot be found in APXs), the rate constants for Compound I formation were measured with peroxoacetic acid and are shown to be 5.9 x 10(4) M(-1) s(-1) for CatPX from A. nidulans and 8.7 x 10(3) M(-1) s(-1) for the Synechocystis enzyme. Compared with o-dianisidine (2.7 6.7 x 10(6) M(-1) s(-1)) and pyrogallol (8.6 x 10(4)-1.6 x 10(5) M(-1) s(-1)), the rate constant for Compound I reduction by ascorbate was extremely low (5.4 x 10(3) M(-1) s(-1) at pH 7.0 and 15 degrees C), in marked contrast to the behaviour of APXs. PMID- 10694067 TI - Catalase activity during C3-CAM transition in Mesembryanthemum crystallinum L. leaves. AB - Treatment with 0.4 mol dm(-3) NaCl caused a C3-CAM shift in Mesembryanthemum crystallinum L. leaves. In parallel to the CAM induction the activity of CAT was significantly decreased. In C3 and in CAM plants CAT activity showed daily fluctuations, with the maximum at the end of the light period. The oscillations of CAT were more pronounced in CAM than in C3 plants. In M. crystallinum CAT activity seems to respond more to CAM induction than to salinity. PMID- 10694068 TI - Laparoscopy at the millennium. PMID- 10694069 TI - A systematic history for the patient with chronic pelvic pain. AB - Chronic pelvic pain is a source of frustration to both the physician and the patient. Physicians have been ill equipped by their training to confront the multifaceted nature of the complaints of patients with chronic pelvic pain. Patients have experienced a repetitive dismissal of their complaints by physicians too busy in their practices to address their problems comprehensively. The approach to the patient with chronic pelvic pain must take into account six major sources of the origin of this pain: 1) gynecological, 2) psychological, 3) myofascial, 4) musculoskeletal, 5) urological, and 6) gastrointestinal. Only by addressing and evaluating each of these components by a very careful history and physical examination and by approaching the patient in a comprehensive manner can the source of the pain be determined and appropriate therapy be administered. This article was developed to provide the clinician with a set of tools and a methodology by which the patient with this complaint can be approached. PMID- 10694070 TI - Combining myoma coagulation with endometrial ablation/resection reduces subsequent surgery rates. AB - BACKGROUND: This study compares results of endometrial ablation alone and in combination with myoma coagulation. Subsequent surgery rates were 38% for ablation alone and 12% for combined therapy. OBJECTIVE: The purpose of this study was to compare hysterectomy rates following various surgical procedures to treat profuse uterine bleeding as well as myomatous uteri. STUDY DESIGN: This is a descriptive study of women who underwent endometrial ablation alone, endometrial ablation with myoma coagulation, or endometrial resection with myoma coagulation to treat profuse uterine bleeding as well as myomatous uterus. From 1986 to 1995, the author performed 52 endometrial ablation procedures; 88 myoma coagulation and endometrial ablation procedures; and 28 myoma coagulations with resection of submucous myomas in patients who were subsequently available for follow-up. Patients were followed up for up to ten years. RESULTS: Of the patients undergoing ablation alone, 20 (38%) of 52 required a second surgery for continued symptoms during a mean follow-up of 47 months. Five of these patients (9.6%) underwent hysterectomy. Of the patients who underwent endometrial ablation plus myoma coagulation (myolysis), 11 (12.5%) of 88 required a repeat surgical procedure during a mean follow-up of 25 months. Five of these patients (5.7%) underwent hysterectomy. Volumetric measurements revealed an average reduction in fibroid volume of 54.5% in this patient group following treatment with a gonadotropin-releasing hormone (GnRH) agonist and combined myoma coagulation and endometrial ablation surgery. Of the 28 patients who underwent myoma coagulation plus resection, five (18%) required a repeat procedure. Of these five, one (4%) required hysterectomy. Fibroid volume in this group was reduced by a mean of 72.6% following administration of a GnRH agonist and combined laparoscopic and hysteroscopic surgery as described. The rate of reoperation was significantly lower among patients receiving endometrial ablation with myoma lysis with or without resection compared with those undergoing endometrial ablation alone (P<0.01). CONCLUSIONS: Myoma coagulation (myolysis), when combined with endometrial ablation among women with symptomatic fibroids and bleeding, reduces all subsequent surgery rates compared with endometrial ablation alone. Myolysis with endometrial resection also results in a reduced need for hysterectomy. PMID- 10694072 TI - The accuracy of gastric insufflation in testing for gastroesophageal perforations during laparoscopic Nissen fundoplication. AB - BACKGROUND: Laparoscopic Nissen fundoplication is an effective technique for the symptomatic relief of the manifestations of gastroesophageal reflux disorder but is associated with a 0.8-1% rate of gastroesophageal perforation. Early detection and repair of these injuries is critical to patient outcome, but occult injuries occur and may be missed. Gastric insufflation technique evaluates the integrity of the gastroesophageal wall after laparoscopic Nissen fundoplication. Gastric insufflation technique involves occlusion of the proximal stomach with a noncrushing bowel clamp while insufflating the submerged gastroesophageal junction. We conducted an animal study to assess the utility of gastric insufflation technique. METHODS: Five pigs (mean weight, 40.4 kg) underwent testing of laparoscopic gastric insufflation technique. In four animals, laparoscopic Nissen fundoplication was performed and then gastroesophageal junction injuries were created (3-5 mm distraction-type wall injuries). Non crushing bowel clamps provided occlusion of the pylorus and then the proximal stomach during gastroesophageal insufflation. The gastroesophageal junction was then submerged. In the fifth animal, gastric insufflation technique was repeated while calibrated injuries were created to determine the smallest detectable injury. An injury was considered detectable if rising air bubbles were noted from the submerged gastroesophageal structures. Maximal luminal pressures needed to detect injuries were recorded with an in-line manometer. RESULTS: In all animals, 5-7 mm injuries of the gastroesophageal junction were easily detected using gastric insufflation technique when the proximal stomach was occluded. When the pylorus alone was occluded, detection of gastroesophageal injuries was inconsistent. Small injuries (<3 mm) of the esophagus were difficult to visualize with pyloric occlusion alone but were consistently detectable with proximal stomach occlusion at pressures less than 20 mm Hg. When the pylorus alone was occluded, the smallest detectable stomach perforation was a 16-gauge needle puncture while applying maximal gastric pressure (40-60 mm Hg) and a 2.5 mm linear injury when generating lower pressures (20 mm Hg). CONCLUSION: Proximal stomach occlusion and insufflation appears to effectively detect esophageal injuries of likely clinical importance (>2.5 mm). Pyloric occlusion and insufflation reliably evaluates the anterior stomach for injury. Gastric insufflation technique is a useful method for detecting gastroesophageal injury after laparoscopic Nissen fundoplication. PMID- 10694071 TI - Laparoscopic Nissen fundoplication in children: a single surgeon's experience. AB - BACKGROUND AND OBJECTIVES: Adult laparoscopic Nissen fundoplication has been steadily growing since its introduction to the United States in the 1990s. Its advantage over the traditional open approach is manifold. Application of laparoscopic fundoplication to children is slowly but surely following this trend. This study evaluates our initial experience with pediatric laparoscopic Nissen fundoplications. PATIENTS AND METHODS: We reviewed the records of 25 consecutive laparoscopic Nissen fundoplications performed by a single surgeon (GS) at our institution in the past three years. The patient ages ranged from 7 months to 18 years (mean, 7 years). All patients had documented gastroesophageal reflux disease. Complications from the reflux included vomiting in 15 patients, failure to thrive in nine, esophagitis in nine, and pulmonary symptoms in six. RESULTS: All Nissen fundoplications were performed laparoscopically without need for conversion to open technique. Blood loss was less than 50 cc in all cases. A tube gastrostomy was concurrently performed in 17. Mean operative time in all cases was 221 minutes. Average postoperative day on which feedings were begun was day 2, with an average resumption of regular feedings on postoperative day 3.5. Average date of discharge was postoperative day 6.8. Complications included difficulty controlling glucose in an insulin-dependent diabetic, and a lost needle, which added an additional hour to the operative time. There were eight admissions to the pediatric intensive care unit, all for observation secondary to their underlying medical problems. There was one postoperative death due to an underlying medical condition. CONCLUSIONS: Laparoscopic Nissen fundoplication is a safe and effective treatment option for children suffering from significant reflux. Time to regular feeding, analgesia requirements and hospital stay are decreased when compared to traditional procedures. Laparoscopic Nissen fundoplication may well become the procedure of choice for pediatric gastroesophageal reflux disease. PMID- 10694073 TI - Laparoscopic surgery for female urinary incontinence: prudence shall prevail. AB - Advances in laparoscopic techniques continue to seek new domains and new indications with the sole objective of providing maximum benefit in a minimally invasive manner. During the last decade, several innovative laparoscopic procedures have evolved for the management of female urinary incontinence. At this juncture, prudence dictates a careful analysis of the principles behind and performance of these procedures so that our treatment recommendations for this common ailment can be based on unbiased scientific pragmatism. In this review, we attempt to analyze the available data and provide constructive criticism and recommendations toward the continued pursuit in this area of development in laparoscopy. PMID- 10694074 TI - Laparoscopic versus open appendicectomy: a comparative study. AB - There are many questions regarding the advantages and disadvantages a videolaparoscopic approach in the treatment of acute appendicitis. The authors present the results of a non-randomized, prospective study with 496 patients admitted between January 1992 and March 1998 by the General Surgery Service of Sao Rafael Hospital Salvador-BA-Brazil. The patients were submitted for appendicectomy by video laparoscopy or by the traditional open method, and variables such as surgery duration, morbidity, mortality, costs, and length of stay (LOS) were compared. The results demonstrate that laparoscopic appendicectomy is a safe alternative for treatment of acute appendicitis; however, there are several disadvantages that gradually must be overcome. PMID- 10694075 TI - Laparoscopic appendectomy and minilaparoscopic approach: a retrospective review after 8-years' experience. AB - BACKGROUND: This is a presentation of our 8-year experience in laparoscopic appendectomy, showing complications and results to determine the advantages and efficacy of laparoscopy. METHODS: We used this technique from December 1990 to December 1998 on 282 consecutive and non-selected patients (169 females and 113 males) with an average age of 24 years (range 5-86 years). All patients were suffering from sub-acute appendicitis or chronic appendicopathies, except for 84 (29.7%) cases of acute appendicitis and 25 (8.9%) cases of gangrenous appendicitis with peritonitis. All patients with suspected appendicitis were evaluated with a laparoscopic exploration. RESULTS: In 39 patients (13.9%), appendectomy was performed along with 19 enucleated or endocoagulated ovarian cysts, 8 adhesiolyses, 6 transperitoneal hernioplasties (4 right and 2 left), 2 cholecystectomies, 2 excisions of a Meckel diverticulum, 1 aspiration and suture of a right tubal pregnancy and 1 electrodesiccation of pelvic endometriosis. Thirty-five patients (12.5%) revealed the presence of a gynecological-type pathology. We performed 2 (0.7%) conversions to open exploration and experienced 6 (2.1%) complications, of which only 1 (0.35%) was a major complication: a delayed hemoperitoneum (1 liter), re-operated elsewhere, the cause of which was not identified. We performed 4 (1.4%) relaparoscopies for retrocecal abscess (three patients with primary gangrenous appendicitis and peritonitis presenting with an abscess in the right iliac fossa and in one patient with widespread intestinal adhesions with primary acute appendicitis). No patient with a diagnosis of a normal appendix developed an intraperitoneal abscess. Mortality was non-existent. The postoperative course, which was subjectively better than in cases operated in the traditional way, was, on an average, 2 days (range 1-18 days) for appendectomies carried out with the traditional laparoscopic technique and 1 day for appendectomies carried out with the minilaparoscopic technique (6 patients). CONCLUSION: We believe that the laparoscopic technique can handle any type of clinical situation, as it can cure several pathologies during the same session with minimal trauma and maximum benefit for the patient. The advantages of a minilaparoscopy approach are based on its low invasiveness and small surgical wounds. PMID- 10694076 TI - Recurrences in laparoscopic incisional hernia repairs: a personal series and review of the literature. AB - OBJECTIVES: Laparoscopic repair of incisional ventral hernias with ePTFE mesh continues to evolve, with variable reporting of surgical techniques and outcomes. This report of 34 cases discusses, with a literature review of laparoscopic incisional hernia repair, specific factors associated with three recurrences. METHOD: Retrospective analysis and review of the literature. RESULTS: Thirty-two patients (16 female, 16 male), underwent 34 laparoscopic repairs: average age-54 years (27-80), average weight-207 lbs (100-300). Nineteen patients (62%) were undergoing first time repairs, 38% were redo cases and 5 cases (14%) involved previous mesh. Operating times averaged 101 minutes (45-220), and average length of stay was 1.9 days (0.6 days excluding 5 patients who required readmission), with 13 patients (38%) being discharged same-day. Two patients developed cellulitis (6%) treated without patch removal. Two enterotomies occurred (6%) both requiring patch removal. Five patients required readmission (14%), and one patient died postoperative day 29 secondary to end-stage liver disease. Three recurrences developed (9%): one secondary to missed enterotomy with reoperation, patch removal and hernia recurrence; one due to omission of suspension suture fixation; and one recurrence developed in a section of the intact old previous incision that extended beyond the original patch. Follow up has averaged 20 months (4-36). CONCLUSIONS: The laparoscopic repair of ventral and incisional hernias utilizing transabdominal placement of ePTFE patch can achieve excellent results with low morbidity in comparison with open surgical approaches. In reviewing the experience of other investigators, adequate fixation of the mesh, extension to cover the entire previous incision and standardizing the placement interval of the sutures are critical to the success of the repair. PMID- 10694077 TI - A cost and profit analysis of hernia surgery. AB - The vast majority of surgeons who are in the active practice of their particular field have little time to evaluate their individual practices from a "business perspective." This fact is critical to the future of any entity that is engaged in the delivery of goods and services. Without such an analysis, few businesses will continue to function in such a manner that ensures the financial viability of that enterprise. We have attempted to accumulate the available data to analyze the practice of surgery as it relates to the cost and profit of hernia repairs. Given this information, it is easily extrapolated into other procedures, open or laparoscopic, that are performed by the general surgeon. The herniorraphy analysis indicates that one cannot hope to generate enough income to rely upon a financially successful business. The information presented should be considered a national average and not specific to an individual practice situation. It is meant to serve as a template for which each surgeon can (and must) evaluate his or her own practice profitability. PMID- 10694078 TI - Criteria and benchmarks for laparoscopic cholecystectomy in a free-standing ambulatory center. AB - BACKGROUND AND OBJECTIVES: Keys to economic survival in an era of decreasing reimbursement include controlling costs and avoiding complications. In an effort to reduce costs, laparoscopic cholecystectomy has been performed with same-day discharge from a hospital setting. The free-standing ambulatory surgery center offers even greater cost savings if safety can be assured. Facility charges, surgical technique and instrument selection influence the costs of the procedure. METHODS: A database was accumulated prospectively on the first 100 laparoscopic cholecystectomies performed in a free-standing ambulatory surgery center to assess costs, logistical constraints, and safety. RESULTS: Laparoscopic cholecystectomies were accomplished in 99 of 100 patients. One patient was suspected of having cancer during laparoscopy and was transferred to a nearby hospital for open cholecystectomy. There were no other postoperative hospitalizations for complications. The fixed facility charge for the procedure was $2,990, and the total costs for all routinely disposable items (gowns, gloves, instruments, and adhesive bandages was $98. The mean OR time was 29 minutes (standard deviation 13.7). CONCLUSIONS: The free-standing ambulatory surgery center is an appropriate facility for an experienced operating team to perform laparoscopic cholecystectomy in selected patients. The surgeon's selection of appropriate energy sources and instruments is essential to complete the operation in a most cost-effective manner. PMID- 10694079 TI - Congenital malformations of the gallbladder and cystic duct diagnosed by laparoscopy: high surgical risk. AB - Congenital anomalies of the gallbladder are rare and can be accompanied by other malformations of the biliary or vascular tree. Being difficult to diagnose during routine preoperative studies, these anomalies can provide surgeons with an unusual surprise during laparoscopic surgery. The presence of any congenital anomaly or the mere suspicion of its existence demands that we exercise surgical prudence, limit the use of electrocoagulation, and ensure that no structure be divided until a clear picture of the bile ducts and blood vessels is obtained. If necessary, perform intraoperative cholangiography to further define the biliary system. However, if the case remains unclear, or if laparoscopy does not provide enough information, open surgery should be considered before undesirable complications occur. PMID- 10694080 TI - Posterior hepatic duct injury during laparoscopic cholecystectomy finally necessitating hepatic resection: case report. AB - A case of bile duct injury during laparoscopic cholecystectomy finally necessitating right hepatic lobectomy is reported to re-emphasize the importance of preoperative and intraoperative assessment of the biliary tree. A 47-year-old Japanese woman underwent laparoscopic cholecystectomy for cholecystolithiasis. On postoperative day 5, fever and right hypochondralgia developed, and CT revealed fluid collection at the right hypochondrium. Percutaneous drainage was performed, and subsequent fistulography revealed a communication of the cystic cavity with the right posterior bile duct, which suggested injury of the aberrant hepatic duct. Conservative therapy, including the adaptation of fibrin glue, was performed, but closure of the fistula and cavity was not obtainable. Finally, a right hepatic lobectomy was performed four months after cholecystectomy. In this case, endoscopic retrograde cholangiopancreatography was unsuccessful preoperatively, and intraoperative cholangiography was not done. This case report re-emphasizes that the preoperative and intraoperative examination of the biliary tree is mandatory to avoid bile duct injury. PMID- 10694081 TI - Laparoscopic placement of peritoneal dialysis catheter (same day dialysis). AB - BACKGROUND AND OBJECTIVE: Peritoneal dialysis (PD) remains the generally accepted method for management of renal failure in chronic and acute renal failure. Despite the rapidly increasing use of continuous ambulatory peritoneal dialysis (CAPD) since its introduction, controversy persists as to the efficacy and exact role of the modality in the treatment of end stage renal failure. The aim of this paper is to present the experience with laparoscopic placement of a peritoneal dialysis catheter and starting the peritoneal dialysis on the same day. METHODS: The laparoscopic placement of a peritoneal dialysis catheter was performed on 11 patients (10 males and 1 female) with an average age of 35 years, over a 12-month period. The procedure was done using two 5 mm abdominal trocars. The precise position of the catheter on the pelvis was ensured laparoscopically. One to two liters exchange dialysis was used for every patient, and no leakage was recorded. RESULTS: The patients tolerated the procedure well. The peritoneal dialysis was started immediately. Patients were discharged after an overnight stay, and PD was carried out routinely. CONCLUSION: The results of laparoscopic placement of a peritoneal dialysis catheter show the following advantages: minimal incision; less surgical trauma; the procedure hastens the early start of peritoneal dialysis and has no complications. PMID- 10694082 TI - Is the standard of care what we think it is? AB - For the most part, gynecologists are actually unaware of the issues involving surrogate versus quality of life outcomes, the "deceptive practice of medicine" and the true incidence of complications as they relate to the standard of care. An anonymous survey of 1958 practicing gynecologists attending seven national symposia revealed a significant number of unreported complications. Clearly, the standard of care (at least with regard to complication risk) is markedly different than has been suggested by the medical literature. Concomitantly, we suggest that physicians need to take a more active role in the policing of our own specialties. PMID- 10694083 TI - 27 months follow-up study of 41 women who underwent laparoscopic supracervical hysterectomy. PMID- 10694084 TI - Tuberculosis control strategies and utilitarianism. PMID- 10694085 TI - Using other people's words. PMID- 10694086 TI - A review of the diagnosis and treatment of smear-negative pulmonary tuberculosis. AB - Recommendations on the management of smear-negative pulmonary tuberculosis (TB) are still based on the behaviour of this disease in populations unaffected by the human immunodeficiency virus (HIV). Studies prior to the HIV epidemic estimated that there were 1.22 cases of smear-negative and extra-pulmonary TB for each smear-positive case. Patients with smear-negative pulmonary TB were found to be less infectious and to have a lower mortality, but a significant proportion (50% 71%) progressed to active disease justifying treatment. Moreover, a wide variety of regimens also proved effective in the treatment of smear-negative disease in HIV-negative patients. The advent of HIV has changed many of these parameters. Countries affected by both HIV and TB have experienced a disproportionate increase in smear-negative disease. While apparently remaining less infectious than smear-positive cases, HIV-positive patients with smear-negative pulmonary TB are generally more immunocompromised, have more adverse drug reactions, and suffer higher mortality rates on treatment. Clinical decision-making has also been complicated because HIV co-infection broadens the differential diagnoses of smear-negative pulmonary TB to include diseases such as Pneumocystis carinii pneumonia (PCP), pulmonary Kaposi's sarcoma, and Gram-negative bacteraemia. Our approach to smear-negative pulmonary TB must therefore adapt to these changed parameters. Management algorithms based on several features (clinical symptoms, response to antibiotic trials, smear investigations, and chest radiography) have been developed to improve case detection. These algorithms must be validated in each locale because their performance will vary depending on numerous local factors such as the regional prevalence of PCP. Alternative methods of specimen collection, such as sputum induction, and processing must be evaluated. National tuberculosis programmes should also consider extending the use of rifampicin based short-course chemotherapy (SCC) to new patients with smear-negative disease. This latter intervention, and the much-needed establishment of additional microscopy and culture facilities, will depend on increased financial and technical support from the international community. PMID- 10694087 TI - Using treatment failure under effective directly observed short-course chemotherapy programs to identify patients with multidrug-resistant tuberculosis. AB - SETTING: Public ambulatory care centers in three districts of northern metropolitan Lima, Peru. OBJECTIVE: To document drug resistance patterns of isolates of Mycobacterium tuberculosis from patients identified as treatment failures under a model tuberculosis (TB) control program based on directly observed, short-course chemotherapy (DOT-SCC). DESIGN: Case series. RESULTS: In a referred, consecutive sample of 173 patients identified as treatment failures on DOT-SCC, 160 (92.5%) had culture-positive TB. Of those 160, 150 (93.8%) had active, pulmonary multidrug-resistant TB (MDR-TB, resistance to at least isoniazid [INH] and rifampicin [RIF]). Sixty of the 150 (40.0%) had isolates resistant to at least INH, RIF, ethambutol (EMB) and pyrazinamide (PZA), the initial first-line empiric treatment regimen used locally. Forty-four (29.3%) had isolates resistant to at least INH, RIF, EMB, PZA and streptomycin (SM), the first retreatment regimen. This series of patients had isolates resistant to a mean of 4.5 of the ten drugs tested. The local profile of multidrug resistance is very different from that obtained from national data from Peru. CONCLUSION: In this setting, treatment failure on DOT-SCC is strongly predictive of active MDR TB. Because of existing local drug resistance patterns in northern Lima, 89.3% of MDR-TB patients identified as treatment failures will receive ineffective therapy with two or fewer secondary TB drugs if they are given the five-drug empiric retreatment regimen endorsed by the World Health Organization. Further short course chemotherapy for these patients would only serve to amplify ominous existing drug resistance patterns. PMID- 10694088 TI - Anti-tuberculosis drug resistance in two clinics in Ecuador. AB - SETTING: Two private hospitals, one in the capital city and one in the eastern rainforest of Ecuador. OBJECTIVE: To document the prevalence of anti-tuberculosis drug resistance in Ecuador in patients who had not received prior treatment and in those who had. DESIGN: Drug resistance was determined using the proportion method with solid medium on the first isolate of Mycobacterium tuberculosis from all patients who attended the two hospitals between 1989 and 1996. Documentation of prior treatment was obtained by patient interview. RESULTS: Resistance was identified in 39 of 161 patients (24%) who had had no prior treatment. Resistance was 14.2% to isoniazid, 11.8% to rifampin and 8.7% to both (multidrug-resistant tuberculosis). Among 60 patients who had received prior treatment, 18 (30%) were resistant to isoniazid, and 14 (23.3%) to rifampin, while multidrug resistance was seen in 10 (16.7%). CONCLUSION: In these populations the prevalence of resistance both in patients with no prior treatment and in patients with prior treatment was ominously high. The initial treatment regimens and patient management in Ecuador should be re-evaluated in an effort to prevent further increases in drug resistance. PMID- 10694089 TI - Rates and risk factors for discontinuation of rifampicin. AB - SETTING: All patients with culture-confirmed, rifampin-susceptible Mycobacterium tuberculosis diagnosed during a 20-month period in New York City, who were started on a rifampin-containing regimen and received > or =60 days of treatment. OBJECTIVE: To identify rates of and reasons for rifampin discontinuation. DESIGN: Retrospective case-control study using surveillance data and medical record reviews. Discontinuation due to thrombocytopenia, creatinine >2.0 mg/dl, bilirubin >2.0 mg/dl or severe reactions (generalized rash, persistent drug fever, or severe interference with methadone metabolism) were defined as appropriate for discontinuation of rifampin. All other reactions were classified as inappropriate. RESULTS: Of 3,520 patients, rifampin was discontinued in 68 (1.9%); of these, 57% had rifampin discontinued unnecessarily. Treatment by an inexperienced provider (adjusted odds ratio [ORadj] 4.0; 95% confidence interval [CI] 1.9-8.5), race (ORadj 3.1; 95%CI 1.4-6.9), history of previous treatment (ORadj 4.8; 95%CI 1.9-12.5), and history of methadone drug treatment (ORadj 12.6; 95%CI 5.3-29.9) were all associated with inappropriate rifampin discontinuation. CONCLUSION: True intolerance was rare, even among those patients infected with the human immunodeficiency virus. Most patients with minor reactions can successfully complete treatment with rifampin, particularly if managed by a physician experienced in the treatment of tuberculosis. PMID- 10694091 TI - Delay in tuberculosis case-finding and treatment in Mwanza, Tanzania. AB - SETTING: Health facilities in Mwanza region, Tanzania. OBJECTIVE: To determine factors responsible for delay from onset of symptoms of pulmonary tuberculosis to initiation of treatment. DESIGN: A cross-sectional descriptive study of 296 smear positive tuberculosis patients. Emphasis was given to periods between 1) onset of symptoms and first consultation to a health facility, and 2) reporting to a health facility and initiation of treatment. RESULTS: Mean total delay was 185 days (median 136), with nearly 90% of this being patient's delay. The mean health system delay was 23 days (median 15), with longer delays in rural health facilities. The mean patient's delay was 162 days (median 120). This delay was significantly longer in rural areas, for patients with lower level of education, for those who first visited a traditional healer, and for patients who had no information on tuberculosis prior to diagnosis. Only 15% of the patients reported to a health facility within 30 days of onset of symptoms. CONCLUSION: There are significant delays in case-finding in Mwanza, Tanzania, with prolonged patient's delay. Facilitation of utilisation of health services, raising awareness of the disease and incorporation of private practice into tuberculosis control could help to reduce these delays. PMID- 10694090 TI - Gender and tuberculosis: a comparison of prevalence surveys with notification data to explore sex differences in case detection. AB - OBJECTIVE: To explore whether lower tuberculosis notification rates among women are due to a reduced access to health care, particularly diagnostic services, for women. METHODS: Age- and sex-specific tuberculosis prevalence rates of smear positive tuberculosis were obtained from tuberculosis prevalence surveys reported to the WHO or published in the literature. Age- and sex-specific notification rates from the same countries in 1996 were used. RESULTS: Prevalence data and notifications from 29 surveys in 14 countries were used. Notification rates varied strongly among countries, but the female/male ratio was below 1 and decreased with increasing age in almost all. The female/male (F/M) prevalence ratios were less than 0.5 in surveys in the South-East Asia and Western Pacific Region, and approximately 1 in the African Region. CONCLUSION: In most countries the F/M sex ratio in prevalent cases was similar or lower than that in notified cases, suggesting that F/M differences in notification rates may be largely due to epidemiological differences and not to differential access to health care. However, available data are limited as the prevalence surveys in Africa were carried out many years ago, and in Asia notification rates may be distorted by a large private sector with deficiencies in notification. PMID- 10694092 TI - Lymph node biopsies in HIV-infected and non-infected children with persistent lung disease. AB - OBJECTIVE: The diagnosis of opportunistic infections in children with persistent lung disease (PLD) who are infected with the human immunodeficiency virus (HIV) is difficult to establish, especially in resource-poor countries. Lymphadenopathy is a frequent associated clinical finding among these children. We evaluated the usefulness of excision lymph node biopsies in determining an aetiological diagnosis in HIV-infected and non-infected children with PLD. METHOD: Forty-five children with PLD and significant lymphadenopathy were subjected to lymph node biopsy. Of these, 27 were HIV-infected. All subjects had excision biopsies; 39 (86.7%) of these cases also underwent fine needle aspiration cytodiagnosis (FNAC) and trucut needle biopsies. RESULTS: Tuberculosis was identified as the final diagnosis in 11 (40.7%) and 12 (66.7%) HIV-infected and noninfected children, respectively. Ancillary investigations (Mantoux, gastric washings) suggested a diagnosis of tuberculosis in eight (72.7%) and eight (66.7%) of the final diagnoses of tuberculosis among HIV-infected and non-infected children, respectively. Lymph node biopsies identified a further three (27.3%) and four (33.3%) more cases of tuberculosis as compared to ancillary investigations among HIV-infected and non-infected groups, respectively. Results of FNAC and trucut biopsy showed good correlation with excision biopsy: 96.4% and 97.4%, respectively. However, adequate samples were obtained in only 23 of 39 FNAC and 33 of 39 trucut biopsies. CONCLUSION: Excision lymph node biopsies form a useful adjunct investigation in children with PLD and generalised lymphadenopathy. The most common disease identified among HIV-infected and non-infected children in Durban, South Africa, is tuberculosis. FNAC and trucut biopsies may also be useful in the evaluation of lymphadenopathy when appropriate specimens are obtained. PMID- 10694093 TI - Yield of gastric lavage and bronchial wash in pulmonary tuberculosis. AB - OBJECTIVE: To determine the yield of acid-fast bacilli (AFB) in gastric lavage and bronchial washing in adult patients clinically and radiologically suspected of pulmonary tuberculosis but who cannot produce sputum. METHODS: Selected adult patients were admitted to the ward; gastric lavage was done for 3 consecutive days after an overnight fast followed by bronchial wash. Specimens were immediately sent to laboratory for AFB direct smear and culture. RESULTS: The yield of AFB in gastric lavage on direct smear was 16/20 (80%) and 12/20 (60%) in the first and second samples, respectively. When combined, 18/20 (90%) were direct smear positive, while the third sample did not increase the yield. The yield of AFB culture in gastric lavage was 6/20 (30%) in both the first and second samples, while the combined results of the first and second samples were 8/20 (40%). The third sample did not increase the yield. In bronchial wash, AFB direct smear was positive in 18/20 (90%), while culture was positive in 14/20 (70%). CONCLUSION: Gastric lavage and bronchial washing are useful methods for the diagnosis of pulmonary tuberculosis in patients who cannot produce sputum. Two gastric lavage specimens are adequate. On comparison, bronchial wash is superior to gastric lavage in culture, but their yield on direct smear is equal. PMID- 10694094 TI - Analysis of circulating immune complexes (CICs) in childhood tuberculosis: levels of specific antibodies to glycolipid antigens and relationship with serum antibodies. AB - BACKGROUND: The presence of specific antiglycolipid antibodies in serum and circulating immune complexes (CIC) in children with tuberculosis was detected in order to evaluate their contribution to the value of serodiagnosis of tuberculosis, as has already been shown in adults. METHODS: ELISAs using the three glycolipids LOS, DAT and PGLTb1 were performed in whole serum and immune complexes from 20 children with tuberculous disease or infection, in seven child contacts, and in 26 children with non-tuberculous disease. The contribution of complexed IgG antibody to the diagnostic values was established for each group. RESULTS: The antibody levels in free serum were higher (P < 0.01) in children with tuberculous disease or infection and in contacts than in controls. By contrast, except for PGLTb1, the IgG antibody levels were higher (P < 0.02) in children with tuberculous disease than in the other groups. The highest contribution of IgG antibody against LOS to the predictive values was shown in children with pulmonary tuberculosis (positive predictive value 1,000, negative predictive value 1,000). In paucibacillary tuberculosis (extra-pulmonary and tuberculous infection) and in contacts, the IgG antibody did not contribute to the sensitivity of the serodiagnosis, where the combination of antigens tested in serum increased the diagnostic yield. The very low levels of IgG antibody in these settings may indicate a different B cell response. CONCLUSION: The detection of immune complexes and IgG antibodies against the three glycolipid antigens is useful as a complementary technique for the serodiagnosis of children with a high probability of pulmonary tuberculosis. PMID- 10694095 TI - Mycobacterium tuberculosis lipid antigens: use of multi-antigen based enzyme immunoassay for free and complex dissociated antibodies. AB - OBJECTIVE: To test the sensitivity and specificity of four lipid antigens of Mycobacterium tuberculosis: BDA-TDA, DAT, SL-I, and PIMs, adsorbed in the same microplate well, to detect reactive IgG by enzyme-immunoassay (EIA) from plain serum (MA-EIA) and dissociated immune complexes (ICMA-EIA). DESIGN: IgG antibodies against four antigens, placed in the same microplate well, were evaluated in serum from 155 tuberculous (TB) cases non-infected with the human immunodeficiency virus (HIV): 78 patients with positive bacilloscopy and culture, 33 patients with positive culture and 44 patients diagnosed by clinical and radiological criteria; and from 211 HIV negative control subjects: 32 patients with other pulmonary diseases, 100 healthy people and 79 close contacts. RESULTS: MA-EIA had an overall sensitivity and specificity of 61% (94/155) and 95% (200/211), respectively. We further examined whether the dissociation of immune complexes increases the number of positive reactions in those initially found to be seronegative (SN). The subset of 112 (76 controls and 36 TB) MA-EIA SN samples tested using ICMA-EIA yielded an overall sensitivity and specificity of 83% and 100%. The ICMA-EIA results improved the overall sensitivity from 61 to 80% without changing specificity. CONCLUSION: These preliminary results suggest that MA-EIA followed by ICMA-EIA, for SN samples, might serve as a fast, cheap, and easy method for the diagnosis of TB in less than 48 hours. PMID- 10694096 TI - Direct drug susceptibility test for tubercle bacilli by the sputum swab culture method. AB - OBJECTIVE: To develop a simple, inexpensive method for testing direct drug susceptibility of tubercle bacilli to isoniazid (INH) and streptomycin (SM) which can be adopted for use even in remote parts of the world. DESIGN: Using 239 smear positive sputum specimens obtained from an equal number of patients, a comparison was made between the direct swab susceptibility test and the standard indirect method for INH and SM using Lowenstein-Jensen (L-J) medium. RESULTS: There was 95% agreement of results for INH by 6 weeks and 90% for SM by 8 weeks; 96% of INH resistant cultures could be detected in 5 weeks and 91% of SM-resistant cultures by 8 weeks. The discrepancies in the two tests were virtually symmetrically distributed at 6 and 8 weeks. The speed and efficiency of detection of resistance by the swab method was also assessed in relation to the standard indirect method. For INH, 96% of the cultures were detected by the fifth week, while 66% were detected as early as 2 weeks and 93% by 4 weeks. With SM, 84% were detected by 4 weeks, 89% by 5 weeks and 91% by 8 weeks. CONCLUSION: This study has indicated the usefulness of the swab method for testing the drug susceptibility of tubercle bacilli. As this method is simple and easy, and does not even require a centrifuge, it has the potential of application even in the remote parts of the world. The material used, Cetavlon (Cetrimide), is inexpensive and easily water soluble, and more importantly, aqueous solutions are self-sterilizing. It should be stressed, however, that the results obtained with this test take the same time as conventional methods, and it can therefore not be considered as a rapid test. PMID- 10694097 TI - Contribution of 'TB clubs' to tuberculosis control in a rural district in Ethiopia. AB - SETTING: A rural district (Estie) in South Gonder, Ethiopia. OBJECTIVE: To describe the contribution of 'TB clubs' (small support groups of patients based on where they live) to the performance of the tuberculosis control programme in Estie District. DESIGN: A descriptive study of the formation of 'TB clubs', their contribution to case-finding and the treatment outcomes before and after formation of the 'TB clubs'. RESULTS: The proportion of actual among expected attendances of tuberculosis patients for follow-up during treatment at health facilities significantly increased (P < 0.001) after the introduction of the TB clubs. Community elders, community health agents and local health workers helped TB clubs to refer tuberculosis suspects, promote treatment adherence and trace defaulters as an integral part of a district tuberculosis programme. The TB clubs referred 181 tuberculosis suspects in the community for investigation, of whom 65% subsequently had a diagnosis of tuberculosis. TB clubs identified 69% of all patients and 76% of new sputum smear-positive pulmonary patients diagnosed in the district. Treatment success rates in new sputum smear-positive, smear-negative and extra-pulmonary tuberculosis patients were 83%, 79% and 81%, respectively. CONCLUSION: The formation of TB clubs contributed to the effective implementation of a district tuberculosis programme. Further evaluation is needed to assess the sustainability and applicability of the approach in other settings. PMID- 10694098 TI - Evaluation of the polymerase chain reaction for the detection of Mycobacterium tuberculosis. AB - The development of nucleic acid-based technologies has improved the sensitivity, specificity and speed of detection of Mycobacterium tuberculosis in clinical samples. Both commercially available and 'in-house' polymerase chain reaction (PCR) systems are in use, and a significant number of reports compare such systems with more traditional diagnostic tools for tuberculosis. Few studies, however, have focused on the reproducibility of the results when submitting a sample batch to PCR in different laboratories, especially in developing countries. Consequently, PCR results obtained from six laboratories in six different Latin American countries for samples reconstituted with defined amounts of M. tuberculosis cells were evaluated. Each laboratory used specific conditions of sample processing, nucleic acid amplification and amplicon detection. Analysis of results allowed large differences in sensitivity and specificity to be observed. We conclude that in its present setting, in-house PCR cannot be used as a single diagnostic tool for tuberculosis, and that special care needs to be taken upon interpretation of results by inclusion of a proper number of positive and negative controls. PMID- 10694099 TI - Predictive values of the ICT Tuberculosis test for the routine diagnosis of tuberculosis in Madagascar. AB - The rapid commercialised ICT Tuberculosis test has been tested in Madagascar for the diagnosis of smear-positive pulmonary (SM+) and extra-pulmonary tuberculosis (EPT), using microscopy, culture and histopathology as reference tests. Specificity was 83.3% and sensitivity 68.2% for SM+ patients and 65.2% for EPT patients. With a prevalence of 22.2% for SM+ patients and 52.4% for confirmed EPT among consulting patients in the National Reference Laboratory, the ICT Tuberculosis assay was not sufficiently predictive for application in the tuberculosis control programme. PMID- 10694100 TI - Ordnung in das Chaos. PMID- 10694101 TI - Study of primary drug resistance of Mycobacterium tuberculosis from two hospitals in Tenerife, Canary Islands, Spain (1990-1997) PMID- 10694102 TI - PAS infusion in treatment of multidrug-resistant tuberculosis. PMID- 10694103 TI - Cardiac responses to progressive exercise in normal children: a synthesis. AB - The cardiac responses to exercise are influenced by a complex interplay of changes in diastolic filling, intrinsic myocardial contractility, heart rate, and ventricular afterload. PURPOSE: To characterize these responses in children, findings are reported from two studies utilizing Doppler echocardiographic assessment of stroke volume and cardiac output during maximal upright and semisupine cycle exercise. METHODS: In study 1, stroke volume, heart rate, and peak aortic velocity were assessed during upright cycle exercise to exhaustion in 39 sixth-grade boys. In study 2, similar methods were used to examine cardiac responses to semisupine exercise with measurement of left ventricular dimensions by two-dimensional echocardiography. RESULTS: The findings support patterns similar to that previously described in adults, with an initial rise in stroke volume reaching a plateau at mild-moderate exercise intensities. CONCLUSIONS: The observations in these two studies also suggest 1) peripheral vasodilatation plays an important role in the early rise in stroke volume, 2) increasing heart rate acts to maintain a stable stroke volume and left ventricular diastolic dimension at high workloads, and 3) improvements in contractility serve to maintain stroke volume as the systolic ejection period shortens. PMID- 10694104 TI - The effects of ankle compliance and flexibility on ankle sprains. AB - PURPOSE: The goal of this study was to examine the influence of changes in subtalar joint flexibility and compliance on ankle sprain occurrence. METHODS: Muscle model driven simulations of 10 subjects performing the landing phase of a side-shuffle movement were performed. The passive flexibility or compliance of the subtalar joint was varied, and each subject-specific simulation was exposed to a set of perturbed floor conditions. RESULTS: Increases in flexibility and compliance both led to an increase in the occurrence of excessive supination, while changes in flexibility had a greater influence. Changes in flexibility or compliance caused only small changes in the occurrence of excessive supination torques. CONCLUSION: These results suggest that increased mechanical laxity does not directly cause an increase in sprain occurrence during side-shuffle movements. PMID- 10694105 TI - Asthma and wheezing among Norwegian elite athletes. AB - PURPOSE: The objectives were to estimate the prevalence of self-reports of asthma and wheezing among Norwegian elite athletes compared with the general population and to estimate the associations between asthma and types of sports, exercise and team level. METHODS: The study population included all Norwegian elite athletes on the national junior and senior teams in 1997 (N = 1620) and a random sample from the general population (N = 1680). The surveys included items for asthma, respiratory symptoms, the history of participation in sports, sports events, and exercise and team level. The associations between the exposure variables and the outcomes adjusting for potential confounding factors were estimated using logistic regression. Crude (c) and adjusted odds ratio (aOR) with 95% confidence interval (CI) are presented. RESULTS: The prevalence of asthma was greater among athletes (10.0%) compared with that in the general population (6.9%) and remained so after controlling for confounders, aOR = 1.5 (95%CI 1.1-2.1). The risk of asthma was highest in sports requiring strength and endurance. This was the case for comparisons between athletes and the general population, aOR = 3.5 (1.6-7.6) for strength and aOR = 2.2 (1.4-3.5) for endurance sports. Comparisons within the sample of athletes using technical sports as the reference category revealed similar results, aOR = 3.0 (1.1-8.0) and aOR = 2.0 (1.0-4.3), respectively. Furthermore, asthma was more common among female than male athletes (aOR 1.7 (1.1 2.7)). Training more than 20 h x wk(-1) was associated with asthma when compared with levels of training less than 10 h x wk(-1) (aOR 1.9 (1.0-4.1)). CONCLUSION: These results indicate that asthma is more common among athletes compared with the general population. Asthma among athletes may define a subgroup of asthma cases for whom etiology is related to extensive exercise. PMID- 10694106 TI - A randomized trial of preexercise stretching for prevention of lower-limb injury. AB - PURPOSE: This study investigated the effect of muscle stretching during warm-up on the risk of exercise-related injury. METHODS: 1538 male army recruits were randomly allocated to stretch or control groups. During the ensuing 12 wk of training, both groups performed active warm-up exercises before physical training sessions. In addition, the stretch group performed one 20-s static stretch under supervision for each of six major leg muscle groups during every warm-up. The control group did not stretch. RESULTS: 333 lower-limb injuries were recorded during the training period, including 214 soft-tissue injuries. There were 158 injuries in the stretch group and 175 in the control group. There was no significant effect of preexercise stretching on all-injuries risk (hazard ratio [HR] = 0.95, 95% CI 0.77-1.18), soft-tissue injury risk (HR = 0.83, 95% CI 0.63 1.09), or bone injury risk (HR = 1.22, 95% CI 0.86-1.76). Fitness (20-m progressive shuttle run test score), age, and enlistment date all significantly predicted injury risk (P < 0.01 for each), but height, weight, and body mass index did not. CONCLUSION: A typical muscle stretching protocol performed during preexercise warm-ups does not produce clinically meaningful reductions in risk of exercise-related injury in army recruits. Fitness may be an important, modifiable risk factor. PMID- 10694107 TI - Anaerobic performance in 5- to 7-yr-old children of low birthweight. AB - PURPOSE: This study was intended to determine whether anaerobic muscle performance is deficient in 5- to 7-yr-old children of extremely low birthweight (ELBW, 500-999 g) and very low birthweight (VLBW, 1000-1499 g). METHODS: Fourteen ELBW and 20 VLBW children were compared with 24 normal birthweight (NBW, >2500 g) term controls. Peak (PP) and mean (MP) muscle power were determined by the Wingate anaerobic test. Bioimpedance analysis and anthropometry were done to assess fat-free mass (FFM) and lean cross-sectional area of the thigh and calf. RESULTS: The ELBW group had significantly lower MP and PP, compared with the VLBW and, in particular, with the NBW group. This lower performance was apparent also when values were corrected for total body mass (MP) and FFM (MP and PP), but not when corrected for cross-sectional area of thigh and calf. CONCLUSION: The lower anaerobic muscle performance in ELBW children may be partly due to their smaller muscle mass, but may also reflect a low percentage of fast-twitch muscle fibers, low muscle phosphagen content, or deficiency in motor control. PMID- 10694108 TI - Influence of intra-oral maxillary sports mouthguards on the airflow dynamics of oral breathing. AB - PURPOSE: Mouthguards worn during sporting competition may influence oral airway flow dynamics and potentially increase airflow resistance during mouth breathing. METHODS: We measured oral airflow resistance (RO) in 10 normal subjects (four men, six women, age 29 +/- 3 yr, mean +/- SEM) wearing two different custom-made maxillary mouthguards. RESULTS: During tidal mouthpiece breathing (jaw position controlled), inspiratory R(O) (at (1.4 L x s(-1)) increased from 0.22 (0.15-0.46) cm H2O x L(-1) x s(-1) (median and interquartile range) to 0.47 (0.24-0.52) cm H2O x L(-1) x s(-1) with mouthguard 1 (general sports mouthguard) and from 0.34 (0.27-0.51) to 0.46 (0.39-0.86) cm H2O x L(-1) x s(-1) (N = 8) with mouthguard 2 (laminated, field hockey mouthguard, both P < 0.05). With oral only mask breathing (jaw position not controlled), inspiratory R(O) (at 0.4 L x s(-1)) increased to 1.02 (0.42-1.57) cm H2O x L(-1) x s(-1) (P < 0.03, compared with mouthpiece) but was variably affected by both mouthguards. At 1.0 L x s(-1), there was a tendency for both mouthguards to increase inspiratory R(O); however, this effect only reached significance for mouthguard 1 during mouthpiece breathing. CONCLUSION: Thus, although maxillary mouthguards do increase R(O) when jaw position is controlled, individual subjects respond differently when in control of mouth opening. This may be related to variable recruitment of compensatory mechanisms (e.g. mouth opening and/or oral airway dilator muscle activity). PMID- 10694109 TI - Acute creatine loading increases fat-free mass, but does not affect blood pressure, plasma creatinine, or CK activity in men and women. AB - Creatine monohydrate (CrM) administration may enhance high intensity exercise performance and increase body mass, yet few studies have examined for potential adverse effects, and no studies have directly considered potential gender differences. PURPOSE: The purpose of this study was to examine the effect of acute creatine supplementation upon total and lean mass and to determine potential side effects in both men and women. METHODS: The effect of acute CrM (20 g x d(-1) x 5 d) administration upon systolic, diastolic, and mean BP, plasma creatinine, plasma CK activity, and body composition was examined in 15 men and 15 women in a randomized, double-blind experiment. Additionally, ischemic isometric handgrip strength was measured before and after CrM or placebo (PL). RESULTS: CrM did not affect blood pressure, plasma creatinine, estimated creatinine clearance, plasma CK activity, or handgrip strength (P > 0.05). In contrast, CrM significantly increased fat-free mass (FFM) and total body mass (P < 0.05) as compared with PL, with no changes in body fat. The observed mass changes were greater for men versus women. CONCLUSIONS: These findings suggest that acute CrM administration does not affect blood pressure, renal function, or plasma CK activity, but increases FFM. The effect of CrM upon FFM may be greater in men as compared with that in women. PMID- 10694110 TI - Estrogen variation and resting left ventricular structure and function in young healthy females. AB - PURPOSE: A potential confounding factor in the interpretation of left ventricular (LV) structural and functional data in female subjects could be menstrual phase or contraceptive use upon assessment. To date no study has addressed this issue. METHODS: Seventeen eumenorrheic (EU; mean +/- SD age = 21 +/- 3 yr) and 14 combined-oral contraceptive pill-using (COC: mean +/- SD age = 21 +/- 3 yr) females volunteered to participate. The EU had stable menstrual cycles and the COC had all been using the same pill preparation for a minimum of 6 months. Echocardiographic examinations occurred during the mid-follicular phase and mid luteal phases in the EU and during mid-consumption and mid-end of withdrawal in the COC. LV structure and function were assessed using M-mode and pulsed-wave Doppler echocardiography. Data were compared via Student t-tests and limits of agreement (LoA) were calculated. RESULTS: Structure and function did not significantly differ between phases of the menstrual cycle or between consumption and withdrawal of oral contraception (P > 0.05). However, there was considerable variance in the LoA between variables. Smaller LoA were reported for those variables directly measured from echocardiograms compared with those from derived data. For example, in a measured variable such as LV internal dimension in diastole, the LoA data represented a variation of +/- 0.4 mm (+/- 1%) between phases. Conversely, data for LV mass, a derived variable, reported LoA values of +/- 15 g (10%) between phases. The LoA were consistent between EU and COC. Variation in both measured and derived variables were within, or close to, accepted limits of measurement or biological error. CONCLUSION: It would seem that in studies assessing LV structure and function in women the influence of menstrual phase or oral contraceptive use is of minor significance. PMID- 10694111 TI - Effect of a neoprene sleeve on knee joint kinesthesis: influence of different testing procedures. AB - PURPOSE: Objectives of this study were to examine the perceived sense of knee joint position during selected test situations, and to evaluate the proposed kinesthetic effect of a neoprene knee sleeve during these test situations. METHODS: Fifty-nine young healthy subjects (39 females and 20 males) attempted to replicate target knee joint angles using active and passive knee extension movements completed in sitting (nonaxially loaded) situations, and during active knee extension movements completed in supine while applying a load of 15% body weight through the long axis of the tibia (axially loaded). The criterion measure used was the absolute difference between target and reproduced angles, averaged over five attempts (Average absolute difference: AAD). RESULTS: A three-way ANOVA (two genders by three test situations by two sleeve conditions), with repeated measures on the last two factors, indicated a significant main effect for test situation and sleeve condition (P < 0.05), but not for gender. There was also a significant test situation by sleeve condition interaction (P < 0.05). Post-hoc analysis indicated that the AAD score during the active nonaxially loaded test situation without the sleeve was significantly greater than AAD scores for all other tests (P < 0.01). CONCLUSIONS: Pre-existing differences in knee joint kinesthesis observed during different contexts of limb movement must be recognized before various interventions, including the effect of knee supports, can be adequately interpreted. Because knee joint position sense was attenuated during voluntary active movement, and because this attenuation was ameliorated by the use of a sleeve, future studies evaluating the kinesthetic effects of knee bracing may benefit from using active movements. However, since the sleeve did not affect performance during the axially loaded test situation, future studies should also evaluate the relationship between tests of knee joint kinesthesis and other more functional tests of neuromuscular performance. PMID- 10694112 TI - Exercise-induced asthma screening of elite athletes: field versus laboratory exercise challenge. AB - PURPOSE: The purpose of this study was to compare a laboratory based exercise challenge (LBC) to a field based exercise challenge (FBC) for pulmonary function test (PFT) exercise-induced asthma (EIA) screening of elite athletes. METHODS: Twenty-three elite cold weather athletes (14 men, 9 women) PFT positive for EIA (FBC screened) served as subjects. Twenty-three gender and sport matched controls (nonasthmatics) were randomly selected to establish PFT reference values for normal elite athletes. Before FBC, athletes completed a medical history questionnaire for EIA symptoms. FBC evaluations consisted of baseline spirometry, actual or simulated competition, and 5, 10, and 15 min postexercise spirometry. PFT positive athletes were evaluated in the laboratory using an exercise challenge simulating race intensity (ambient conditions: 21 degrees C, 60% relative humidity). PFT procedures were identical to FBC. RESULTS: 91% of PFT positive and 48% of PFT normal athletes reported at least one symptom of EIA, with postrace cough most frequent. Baseline spirometry was the same for PFT positives and normal controls. Lower limit reference range (MN - 2 SD) of FEV1 for controls suggests that postexercise decrements of greater than approximately 7% indicate abnormal airway response in this population. Exercise time duration did not effect bronchial reactivity; 78% of FBC PFT positives were PFT normal post-LBC. CONCLUSION: Self-reported symptoms by elite athletes are not reliable in identifying EIA. Reference range criterion for FEV1 decrement in the elite athlete postexercise contrasts current recommended guidelines. Moreover, a large number of false negatives may occur in this population if EIA screening is performed with inadequate exercise and environmental stress. PMID- 10694113 TI - Cytokine hypothesis of overtraining: a physiological adaptation to excessive stress? AB - Overtraining syndrome (OTS) is a condition wherein an athlete is training excessively, yet performance deteriorates. This is usually accompanied by mood/behavior changes and a variety of biochemical and physiological alterations. Presently, there is no global hypothesis to account for OTS. The present paper will attempt to provide a unifying paradigm that will integrate previous research under the rubric of the cytokine hypothesis of overtraining. It is argued that high volume/intensity training, with insufficient rest, will produce muscle and/or skeletal and/or joint trauma. Circulating monocytes are then activated by injury-related cytokines, and in turn produce large quantities of proinflammatory IL-1beta, and/or IL-6, and/or TNF-alpha, producing systemic inflammation. Elevated circulating cytokines then co-ordinate the whole-body response by: a) communicating with the CNS and inducing a set of behaviors referred to as "sickness" behavior, which involves mood and behavior changes that support resolution of systemic inflammation: b) adjusting liver function, to support the up-regulation of gluconeogenesis, as well as de novo synthesis of acute phase proteins, and a concomitant hypercatabolic state; and c) impacting on immune function. Theoretically, OTS is viewed as the third stage of Selye's general adaptation syndrome, with the focus being on recovery/survival, and not adaptation, and is deemed to be "protective," occurring in response to excessive physical/physiological stress. Recommendations are made for potential markers of OTS, based on a systemic inflammatory condition. PMID- 10694114 TI - Blood volume: importance and adaptations to exercise training, environmental stresses, and trauma/sickness. AB - This paper reviews the influence of several perturbations (physical exercise, heat stress, terrestrial altitude, microgravity, and trauma/sickness) on adaptations of blood volume (BV), erythrocyte volume (EV), and plasma volume (PV). Exercise training can induce BV expansion: PV expansion usually occurs immediately, but EV expansion takes weeks. EV and PV expansion contribute to aerobic power improvements associated with exercise training. Repeated heat exposure induces PV expansion but does not alter EV. PV expansion does not improve thermoregulation, but EV expansion improves thermoregulation during exercise in the heat. Dehydration decreases PV (and increases plasma tonicity) which elevates heat strain and reduces exercise performance. High altitude exposure causes rapid (hours) plasma loss. During initial weeks at altitude, EV is unaffected, but a gradual expansion occurs with extended acclimatization. BV adjustments contribute, but are not key, to altitude acclimatization. Microgravity decreases PV and EV which contribute to orthostatic intolerance and decreased exercise capacity in astronauts. PV decreases may result from lower set points for total body water and central venous pressure, while EV decreases may result from increased erythrocyte destruction. Trauma, renal disease, and chronic diseases cause anemia from hemorrhage and immune activation which suppresses erythropoiesis. The re-establishment of EV is associated with healing, improved life quality, and exercise capabilities for these injured/sick persons. PMID- 10694115 TI - Left ventricular morphology in junior and master resistance trained athletes. AB - PURPOSE: The objective of this cross-sectional investigation was to assess the effects of short (< 5 yr) versus long-term (> or = 18 yr) resistance training (RT) on left ventricular (LV) dimensions and mass. METHODS: The subjects for this study were 20 elite male powerlifters (8 junior athletes [JA], mean +/- SD, age: 21.1 +/- 1.2 yr and 12 master athletes [MA], age: 46.0 +/- 5.5 yr) and 19 age matched male controls (8 young controls [YC], age: 21.8 +/- 2.8 yr and 11 middle aged controls [MAC], age: 46.8 +/- 4.4 yr). Two-dimensionally guided transthoracic M-mode echocardiograms were performed at rest to quantify LV systolic and diastolic cavity dimension (LVIDs and LVIDd, respectively), ventricular septal wall thickness (VST), posterior wall thickness (PWT), LV mass (LVM), and LV systolic function as measured as fractional shortening (FS). RESULTS: Short- or long-term RT was not associated with a significant alteration in LVIDd (JA: 53.2 +/- 4.5 mm, YC: 52.1 +/- 3.7 mm, MA: 53.0 +/- 5.1 mm, MAC: 51.8 +/- 4.4 mm), LVIDs (JA: 33.5 +/- 4.8 mm, YC: 32.9 +/- 3.4 mm, MA: 33.0 +/- 4.4 mm, MAC: 31.4 +/- 3.7 mm), VST (JA: 9.4 +/- 0.9 mm, YC: 9.4 +/- 0.9 mm, MA: 9.4 +/- 1.6 mm, MAC: 9.7 +/- 0.9 mm), PWT (JA: 9.2 +/- 0.9 mm, YC: 9.4 +/- 0.9 mm, MA: 9.0 +/- 1.1 mm, MAC: 9.5 +/- 1.0 mm), LVM (JA: 184.6 +/- 36.1 g, YC: 179.0 +/- 26.5 g, MA, 183.3 +/- 58.1 g, MAC: 184.1 +/- 38.1 g), or FS (JA: 0.37 +/- 0.1%, YC: 0.37 +/- 0.05%, MA: 0.38 +/- 0.1%, MAC: 0.40 +/- 0.04%). CONCLUSIONS: These findings suggest that short or long-term RT as performed by elite junior and master powerlifters does not alter LV morphology or systolic function. PMID- 10694116 TI - Physical activity, cardiorespiratory fitness, and the primary components of blood viscosity. AB - PURPOSE: The relationship of both self-reported leisure time physical activity (LTPA) and predicted maximum oxygen consumption (VO2max) with plasma viscosity and hematocrit (Hct) concentration was examined within a cross-sectional sample of employed middle-aged men. METHODS: Analyses were performed on a subsample of nonsmoking men who completed a preventive medical assessment between 1992-1996. RESULTS: Among nonsmokers the mean age-adjusted levels of plasma viscosity (N = 590) and Hct concentration (N = 632) were significantly lower with higher Physical Activity Index (PAI) categories (P = 0.001 and P = 0.01, respectively). Following adjustment for conventional IHD risk factors and blood leukocyte count, a significant inverse relationship remained for Hct (P = 0.044) but not plasma viscosity. Mean age-adjusted plasma viscosity and Hct concentration also showed a significant decrease with higher quartiles of predicted VO2max (mL x kg(-1) x min(-1))(P = < 0.00005 and P = 0.0004, respectively). Following adjustment for all confounding variables mean plasma viscosity and Hct concentration remained significantly lower with higher quartiles of predicted VO2max (mL x kg(-1) x min( 1))(P = < 0.00005 and P = 0.047). CONCLUSIONS: These data confirm the inverse relationship between LTPA and/or predicted VO2max with plasma viscosity and Hct concentration within nonsmoking middle-aged men of high socioeconomic status. PMID- 10694117 TI - Effects of prior exercise on eccentric exercise-induced neutrophilia and enzyme release. AB - PURPOSE: The purpose of this study was to determine the effects of prior exercise on changes in circulating neutrophils, neutrophil activation, and myocellular enzymes following a standardized bout of eccentric exercise. METHODS: Twenty-four male volunteers were randomized into three groups (N = 8). Group C performed 10 sets of 10 eccentric contractions of the quadriceps muscles with both legs (100% of the concentric IRM). Group D and Group F exercised for 2 h at 56%VO2max on a cycle ergometer followed by a similar bout of eccentric contractions. Group F also received 7.5 mL x kg(-1) of a carbohydrate-electrolyte beverage every 30 min during the submaximal exercise, whereas group D received no fluid. RESULTS: Body weight remained unchanged in groups C and F and decreased in group D by 1.56 +/- 0.34 kg. Ultrastructural Z-Band damage increased three-fold following exercise and remained elevated 3 d after exercise but was not different among groups. Circulating neutrophils were elevated more in group D compared with those in group C immediately after the exercise or rest period, and this difference persisted 3 h after the eccentric exercise. Serum lactoferrin concentrations increased 3.3-fold after exercise in all groups (P < 0.01). Creatine kinase levels (CK) rose in all subjects, with subjects in Group F and D having a significantly greater rise in CK after exercise compared with those in group C. CONCLUSIONS: These data indicate that submaximal exercise followed by a bout of eccentric exercise results in similar amounts of myofibrillar injury with a larger neutrophil response and CK release. PMID- 10694118 TI - Induction of mitochondrial stress proteins following treadmill running. AB - PURPOSE: The purpose of this investigation was to examine the relationship between the expression of HSP60 and GRP75 and the oxidative potential of skeletal muscle as assessed by the citrate synthase activity following endurance training to sedentary controls. METHODS: Female Wistar rats were assigned to one of two groups: sedentary controls (N = 8) or endurance trained (N = 9). Endurance trained rats were run 60 min x d(-1) at 27 m x min(-1) up a 10% incline 6 d x wk( 1) for 8 wk on a motor-driven treadmill. RESULTS: Training produced a 47% increase in citrate synthase activity along with a 103% increase in the expression of HSP60 and a 105% increase in the expression of GRP75 in plantaris muscle. In addition, there was a significant correlation between the citrate synthase activity and expression of HSP60 found in plantaris muscle. CONCLUSIONS: These findings are consistent with the hypothesis that the adaptive response to treadmill running may require elevations in the expression of HSP60 and GRP75 to support protein import and folding. PMID- 10694119 TI - Plasma creatine kinase responses of 18- to 30-yr-old African-American men to eccentric exercise. AB - PURPOSE: The primary aim was to describe the response of plasma creatine kinase (CK) activity in a sample of African-American men after a bout of eccentric exercise. The study also described signal intensity changes detected by MR in the musculature of the right lower leg. METHODS: Subjects were 20 male volunteers of African descent (age = 24 +/- 4 [mean +/- SD] yr). Each walked backward for 60 min at 3 km x h(-1) down a 23% grade. Venous blood was sampled before exercise, immediately (0 d) after and 1, 2, 4, and 7 d after exercise for plasma CK assay. Soreness in the plantar flexor muscles was evaluated in 18 subjects at selected times during the 7 d postexercise. Injury to the plantar flexor muscles was evaluated by magnetic resonance imaging (MRI). RESULTS: The subjects had high baseline plasma CK activity (187 +/- 127 IU x L(-1); 163 +/- 70 IU x L(-1) with one outlier excluded) compared with typical clinical norms. As a group, CK activity was increased (P < 0.05) 4 d (980 +/- 1331 IU L(-1)) and 7 d (1022 +/- 1031 IU L(-1)) postexercise, compared with preexercise. Eleven (55%) of the subjects had large, delayed increases in plasma CK activity ("hyperresponses"). As a group, the plasma CK response was similar to responses of comparison Caucasian subjects. All subjects reported delayed muscle soreness; there was no association between soreness and plasma CK. Every subject showed MRI evidence of injury to plantar flexor muscles postexercise but varying in degree and time course. Plasma CK activity correlated to MRI signal intensity (rho = 0.445). CONCLUSION: Results suggest that changes in plasma CK activity and skeletal muscle injury in African-American men after eccentric exercise do not differ from the responses of Caucasians. PMID- 10694120 TI - Creatine enhances oxygen uptake and performance during alternating intensity exercise. AB - PURPOSE: The main purpose of the present study was to measure the total oxygen consumed, accumulation of blood metabolites, and performance during alternating intensity exercise before and after a period of creatine (Cr) loading in well trained humans. METHODS: Fourteen males were randomly assigned to two groups of seven males and were tested before and after 5 d of placebo (PL) or Cr monohydrate (CR) loading (20 g x d(-1)). Oxygen uptake was measured using a breath-by-breath system during bicycle exercise alternating every 3 min between bouts at 30%(-30%) and 90% (-90%) of the maximal power output to exhaustion. Blood samples were also obtained at rest, before the end of each cycling load, at exhaustion, and 5-min postexercise. RESULTS: The oxygen consumed during 1-90% (5.08 +/- 0.39 L) and 2-90% (5.32 +/- 0.30 L) was larger after CR (5.67 +/- 0.34 and 5.78 +/- 0.35 L, P < 0.01 and P < 0.05, respectively). Blood ammonia accumulation at the end of 1-90% (23.1 +/- 6.5 micromol x L(-1)) and 3-30% (64.7 +/- 15.2 micromol x L(-1)) was lower after CR (P < 0.05), whereas plasma uric acid accumulation was lower at exhaustion (P < 0.05) and 5-min postexercise (P < 0.01). Time to exhaustion increased (P < 0.05) from 29.9 +/- 3.8 to 36.5 +/- 5.7 min after CR, whereas it remained the same after PL. CONCLUSIONS: The results indicate that Cr feeding increases the capacity of human muscle to perform work during alternating intensity contraction, possibly as a consequence of increased aerobic phosphorylation and flux through the creatine kinase system. PMID- 10694121 TI - Effects of abstinence from cigarette smoking on the cardiorespiratory capacity. AB - PURPOSE: The purpose of this study was to determine the effect of 6 or 7 d of abstinence from cigarette smoking on the cardiorespiratory capacity of young men. METHODS: The subjects were 11 male volunteers, mean age 20.5 yr. Their mean smoking duration and cigarettes smoked per day were about 3.5 yr and 20.5 cigarettes, respectively. On the first day of the study, the subjects' physical characteristics, pulmonary functions, and maximal oxygen debt predicted by bicycle ergometer exercise were measured. The subjects' heart rate at rest (HRrest), blood pressure, venous blood analysis and maximal oxygen intake (VO2max) in a treadmill exercise test were measured on the second day. From the time of the second-day measurements, the subjects abstained from cigarette smoking. On the eighth and ninth days of the study, the same measurements were taken as the measurements of the first and second days, respectively. RESULTS: The changes between before and after abstinence from smoking were as follows. The plasma cotinine had almost disappeared after the 7 d of abstinence from smoking. Pulmonary functions did not show a significant improvement. Although there were no significant changes in the HRrest or systolic or diastolic blood pressure, the pulse pressure was significantly increased (P < 0.05). Although the blood components were not changed significantly, the PO2, O2 saturation, HCO3, and CO2 concentration were significantly increased. The predicted maximal oxygen debt was not significantly decreased. Although the VO2max and maximal ventilation volume were not changed, the exercise time was significantly prolonged (P < 0.001). The heart rate during the treadmill exercise at almost all stages decreased significantly; however, the maximal heart rate did not change. CONCLUSION: In conclusion, exercise performance was improved by the 7 d of abstinence from smoking. PMID- 10694122 TI - The association between vigorous physical activities and fat deposition in male adolescents. AB - PURPOSE: The aim of this study was to investigate the association between participation in vigorous physical activities (VPA) and indicators of adiposity and fat distribution in male adolescents. METHODS: Subjects were classified on the basis of the number of reported 15-min periods over 3 d during which VPA were performed. RESULTS: Body weight, fat mass, body mass index, sum of six subcutaneous skinfolds, trunk and extremity skinfolds, and trunk to extremity skinfolds ratio (T/E ratio) were all significantly and inversely related to a higher VPA participation. Moreover, T/E ratio was significantly lower in the subgroup reporting a mean of 7.8 15-min periods of activity for 3 d (equivalent to a mean of 39 min x d(-1)) compared with subjects reporting no participation. This decrease in T/E ratio was more pronounced in subjects reporting a mean of 13.6 15-min periods of VPA for 3 d. CONCLUSION: These observations reinforce the idea that excess abdominal fat accumulation can be prevented by regular participation in vigorous physical activities. PMID- 10694123 TI - Effect of mild dehydration on the lactate threshold in women. AB - PURPOSE: The purpose of this investigation was to examine the effects of dehydration on the lactate threshold and performance time to exhaustion in women. METHODS: Seven moderately trained women (age = 23.6 +/- 1.6 yr) performed two graded exercise tests on separate occasions, once in a normally hydrated state (HY) and once in a dehydrated state (DE). Dehydration was achieved by a 45-min submaximal exercise the evening before testing, followed by a 12-h period of fluid restriction. VO2, VCO2, V(E), R-values, blood lactate, and catecholamine concentrations were measured at baseline and during each workload. Plasma volume and plasma osmolality were also determined. Body weight dropped significantly for the dehydrated trial (2.6 +/- 0.7%). RESULTS: There was a corresponding decrease in plasma volume measured (3.5 +/- 2.6%). The VO2max (3.1 +/- 0.3 L x min(-1) HY; 3.0 +/- 0.1 L x min(-1) DE) obtained was not significantly different between the hydration and dehydration trial. Plasma norepinephrine, epinephrine, and lactate concentrations were not significantly different at baseline or maximum intensity although epinephrine concentrations were higher for the dehydrated trial during submaximal workloads. Lactate concentrations were highly correlated with epinephrine (r = 0.95 HY; r = 0.97 DE). The lactate threshold occurred at a significantly lower relative percent of VO2max for the dehydrated trial (72.2 +/- 1.1% HY; 65.5 +/- 1.8% DE) as well as a lower absolute power output when compared with that in the hydrated trial. There was a significant decrease in time to exhaustion for the dehydrated trial (17.3 +/- 0.7 min HY; 16.3 + 0.7 min DE). Time to exhaustion for the dehydrated trial was correlated with the % VOmax at which the lactate threshold occurred (r = 0.74). CONCLUSIONS: These data indicate that low levels of dehydration induced a shift in the lactate threshold, in part because of elevated epinephrine concentrations. This shift may have been one cause for the decrease in time to exhaustion for the dehydrated trial. PMID- 10694124 TI - Postactivation potentiation in endurance-trained male athletes. AB - PURPOSE: The purpose of the study was to determine whether postactivation potentiation (PAP) was enhanced in the trained muscles of male endurance athletes. METHODS: Triathletes (TRI), distance runners (RUN), active controls (AC), and sedentary control subjects (SED) (N = 10 per group) performed 10-s maximal isometric contractions (MVC) of the elbow extensor and ankle plantarflexor muscles. Maximal twitch contractions were evoked (percutaneous stimulation) before and during a 5-min period after the MVC. PAP was measured as the percentage change in peak twitch torque post-MVC. RESULTS: TRI, who train both upper and lower limb muscles, had enhanced (relative to SED) PAP in both elbow extensor and plantarflexor muscles. In RUN, who train only the lower limbs, enhanced PAP was restricted to the plantarflexors. AC, whose main activity was upper and lower limb weight training, also had enhanced PAP in both muscle groups, although the enhancement in the plantarflexors was not as great as in TRI and RUN. CONCLUSION: PAP is enhanced in endurance athletes. Enhanced PAP may counteract fatigue during endurance exercise. The mechanism(s) responsible for the enhanced PAP remain to be determined. PMID- 10694125 TI - Muscular strength and physical function. AB - PURPOSE: The purpose of this study was to evaluate the potential association of muscular strength and endurance at baseline with the prevalence of functional limitations at follow-up. METHODS: Study participants were 3,069 men and 589 women (30-82 yr) who received a clinical examination including a strength evaluation at the Cooper Clinic between 1980 and 1989 and responded to a 1990 mail-back survey. Participants also had to achieve at least 85% of their age predicted maximal heart rate on a maximal exercise treadmill test and have no history of heart attack, stroke, diabetes, high blood pressure, cancer, or arthritis at their first visit. A strength index composite score (0-6) was calculated using age- and sex-specific tertiles from bench press, leg press, and sit-up tests. Those scoring 5 or 6 were categorized in the high strength group. Functional health status was assessed by responses to questions about the participant's ability to perform light, moderate, and strenuous recreational, household, daily living, and personal care tasks. RESULTS: After an average follow-up of 5 yr, 7% of men and 12% of women reported at least one functional limitation. A logistic regression model including age, aerobic fitness, body mass index, and new health problems at follow-up found that, relative to those with lower levels of strength, the odds of reporting functional limitations at follow up in men and women categorized as having higher levels of strength were 0.56 (95%CI = 0.34, 0.93) and 0.54 (95%CI = 0.21, 1.39), respectively. CONCLUSIONS: These findings, if replicated in other populations, suggest that maintenance of strength throughout the lifespan may reduce the prevalence of functional limitations. PMID- 10694126 TI - Leisure-time physical activities and their relationship to cardiorespiratory fitness in healthy men and women 18-95 years old. AB - PURPOSE: We examined leisure-time physical activities (LTPA) and their contribution to peak oxygen consumption (VO2) in healthy men (N = 619) and women (N = 497) aged 18-95 yr (mean 51 +/- 17) who were participants of the Baltimore Longitudinal study of Aging. METHODS: Calculations of LTPA were based on the average self-reported time spent performing 97 activities and converted into MET min x 24 h(-1). The activities were divided into three levels of LTPA based on absolute intensity. Peak VO2 was determined from a maximal treadmill exercise test. RESULTS: Total LTPA was inversely related to age in both sexes (r = -0.26, P < 0.0001 in men and r = -0.23, P < 0.0001 in women), mediated primarily by less high-intensity activities in older subjects, with only minor differences in moderate- and low-intensity activities across age. Peak VO2 correlated positively with LTPA; the correlations were strongest for high-intensity LTPA (r = 0.33 in men and 0.27 in women, each P < 0.0001), intermediate for moderate-intensity activity (r = 0.12, P < 0.004 in men and r = 0.17, P < 0.0001 in women) and minimal for low-intensity activity (r = 0.08, P = 0.05 in men and r = 0.06, P = 0.20 in women). On univariate analysis, total LTPA accounted for 12.9% of peak VO2 variance for men and 10.6% for women. By multivariate analysis, LTPA independently accounted for 1.6% of the peak VO2 variance in men and 1.8% in women after controlling for age and body mass index. CONCLUSIONS: In healthy adults across a broad age range, LTPA is a relatively minor independent contributor to aerobic capacity. PMID- 10694127 TI - Using objective physical activity measures with youth: how many days of monitoring are needed? AB - PURPOSE: The purpose of this study was to establish the minimal number of days of monitoring required for accelerometers to assess usual physical activity in children. METHODS: A total of 381 students (189 M, 192 F) wore a CSA 7164 uniaxial accelerometer for seven consecutive days. To examine age-related trends students were grouped as follows: Group I: grades 1-3 (N = 92); Group II: grades 4-6 (N = 98); Group III: grades 7-9 (N = 97); Group IV: grades 10-12 (N = 94). Average daily time spent in moderate-to-vigorous physical activity (MVPA) was calculated from minute-by-minute activity counts using the regression equation developed by Freedson et al. (1997). RESULTS: Compared with adolescents in grades 7 to 12, children in grades 1 to 6 exhibited less day-to-day variability in MVPA behavior. Spearman-Brown analyses indicated that between 4 and 5 d of monitoring would be necessary to a achieve a reliability of 0.80 in children, and between 8 and 9 d of monitoring would be necessary to achieve a reliability of 0.80 in adolescents. Within all grade levels, the 7-d monitoring protocol produced acceptable estimates of daily participation in MVPA (R = 0.76 (0.71-0.81) to 0.87 (0.84-0.90)). Compared with weekdays, children exhibited significantly higher levels of MVPA on weekends, whereas adolescents exhibited significantly lower levels of MVPA on weekends. Principal components analysis revealed two distinct time components for MVPA during the day for children (early morning, rest of the day), and three distinct time components for MVPA during the day for adolescents (morning, afternoon, early evening). CONCLUSIONS: These results indicate that a 7 d monitoring protocol provides reliable estimates of usual physical activity behavior in children and adolescents and accounts for potentially important differences in weekend versus weekday activity behavior as well as differences in activity patterns within a given day. PMID- 10694128 TI - Characteristics and etiology of exercise-related transient abdominal pain. AB - PURPOSE AND METHODS: In order to investigate the characteristics of the exercise related transient abdominal pain (ETAP) commonly referred to as "stitch," a questionnaire was administered to a total of 965 regular sporting participants involved in six sports. RESULTS: The percentage of respondents claiming to have experienced ETAP within the past year for the respective sports were: running (69%, N = 439), swimming (75%, N = 103), cycling (32%, N = 76), aerobics (52%, N = 126), basketball (47%, N = 121), and horse riding (62%, N = 100). ETAP appears to be most prevalent in activities that involve repetitive torso movement, either vertical translation or longitudinal rotation. ETAP appears to be a single condition, common in its manifestation to most sufferers, and was described by respondents as a well-localized pain (79%), mostly experienced in the right or left lumbar regions of the abdomen (78%). The sensation of ETAP may be related to the severity of pain with less intense ETAP being described as cramping, aching, or pulling, and greater severity ETAP as sharp or stabbing in nature. Fourteen percent of respondents indicated that they experience shoulder tip pain (STP), which being the diaphragmatic-referred site could suggest irritation of the diaphragm. Respondents claiming to have experienced ETAP were more likely to report STP (r = 0.14, P < 0.01). CONCLUSIONS: The findings of the present study provide perspective on previously suggested etiologies of ETAP, which include diaphragmatic ischemia and stress on the visceral "ligaments," and form the basis for examining alternative etiologies such as cramp of the musculature and irritation of the parietal peritoneum. PMID- 10694129 TI - Effect of bright light on cycling performance. AB - PURPOSE: The purpose of this study was to examine the effect of exposure to three levels of light intensity during cycling on average power output during an all out 20-min bout of cycle ergometry. METHODS: Twelve male competitive cyclists, with a mean age of 24.3 +/- 3.2 yr, were tested on four separate occasions. During each test session, participants were instructed to produce the highest possible average power output during a 20-min bout of cycle ergometry. Session 1 served as a adaptation trial and was conducted in normal room lighting (approximately 250 lux). In subsequent sessions each participant exercised while sitting in front of a light box. Light intensity was experimentally manipulated through the use of sunglasses that provided high, medium, and low light filtration. Light condition order was counterbalanced. Measurements of heart rate, muscle pain, local perceived exertion, metabolism, and alertness were made during exercise. Mood was assessed before and following exercise with the POMS BI. RESULTS: There were no statistically significant differences in the average total power output in the 1,411 lux (274.9 +/- 21.8 W), 2,788 lux (274.4 +/- 20.5 W), and 6,434 lux (270.3 +/- 19.8 W) light conditions. The difference between the average power output in the brightest and least bright condition (4.6 W) was approximately one-fourth of a pooled SD (d = 0.22). There were no significant differences among the conditions in alertness, leg muscle pain, perceived exertion, heart rate, VO2, or mood responses to the exercise. CONCLUSION: It is concluded that exposure to bright light resulting in estimated retinal exposures ranging from 1,411 to 6,434 lux did not have a large or systematic influence on the performance of 20 min of maximal cycle ergometry. PMID- 10694130 TI - Perceived submaximal force production in young adult males and females. AB - Clinical treatment in physical rehabilitation routinely uses perceived relative forces. PURPOSE: This study used psychophysical methods to quantify subjects' errors during submaximal muscular force production. METHODS: A sample of young adult (aged 23 +/- 3 yr) females (N = 60, 62 +/- 14 kg) and males (N = 50, 72 +/- 13 kg) performed a chest press on a hydraulic dynamometer with which they were unfamiliar. In four consecutive presses with a 2-min rest interval between each press, the subjects were asked to produce a force in the order of 25%, 50%, 75% of their maximal force, and a final maximal press. Pilot data (N = 10) indicated good reliability (r(xx) > 0.80) for the protocol. RESULTS: The rs between perceived force production and the desired production were 0.76 (P < 0.001) for males and 0.75 (P < 0.001) for females. The exponent for the power function between the perceived and desired forces was 1.12 for males and 1.03 for females. The total error ranged from 2.82 kg x m (males) to 1.22 kg x m (females). The rs and the logarithmic matching of perceived and desired force indicated a linear relationship that is consistent with Borg's range model, which has proposed that perceptual intensities of force for different people are approximately set equal at a subjective maximum. CONCLUSION: Many young healthy subjects can produce relative muscular forces with good accuracy. However, some subjects will provide very inaccurate forces that might affect outcomes in rehabilitation or physical training. PMID- 10694131 TI - Children's OMNI scale of perceived exertion: mixed gender and race validation. AB - PURPOSE: The newly developed Children's OMNI Scale of Perceived Exertion (category range: 0 to 10) was validated using separate cohorts of female and male, African American and white subjects. Each of the four cohorts contained 20 clinically normal, nonobese children, 8-12 yr of age. METHODS: A cross-sectional, perceptual estimation paradigm using a single multi-stage cycle ergometer test protocol was used. Oxygen uptake (VO2; mL x min(-1)), heart rate (HR; beats x min(-1)) and ratings of perceived exertion for the overall body (RPE-Overall), legs (RPE-Legs), and chest (RPE-Chest) were determined at the end of each continuously administered 3-min power output (PO) (i.e., 25, 50, 75, and 100 W) test stage. RESULTS: The range of responses over the four POs for all cohorts was VO2: 290.8 to 1204.0 mL x min(-1); HR: 89.2 to 164.4 beats x min(-1); and RPE Overall, RPE-Legs, and RPE-Chest: 0.85 to 9.1. First-order correlation and linear regression analyses were performed for each cohort separately and the total sample using a repeated measures paradigm over the four POs. For all correlation/regression paradigms RPE-Overall, RPE-Legs, and RPE-Chest distributed as a positive linear function of both VO2 and HR; r = 0.85 to 0.94; P < 0.01. Differences between RPE-Overall, RPE-Legs, and RPE-Chest were examined with ANOVA for the repeated measures paradigm. RPE-Legs was higher (P < 0.01) than RPE-Chest and RPE-Overall at 25, 50, 75, and 100 W. RPE-Chest did not differ from RPE Overall at 25 and 50 W but was lower (P < 0.01) than RPE-Overall at 75 and 100 W. CONCLUSION: The psycho-physiological responses provide validity evidence for use of the Children's OMNI Scale over a wide range of dynamic exercise intensities. PMID- 10694132 TI - Quadriceps EMG/force relationship in knee extension and leg press. AB - PURPOSE: This study compared the relationship between surface electromyographic (EMG) activity and isometric force of m. quadriceps femoris (QF) in the single joint knee extension (KE) and the multi-joint leg press (LP) exercises. METHODS: Nine healthy men performed unilateral actions at a knee angle of 90 degrees at 20, 40, 60, 80, and 100% of maximal voluntary contraction (MVC). EMG was measured from m. vastus lateralis (VL), m. vastus medialis (VM), m. rectus femoris (RF), and m. biceps femoris (BF). RESULTS: There were no differences in maximum EMG activity of individual muscles between KE and LP. The QF EMG/force relationship was nonlinear in each exercise modality. VL showed no deviation from linearity in neither exercise, whereas VM and RF did. BF activity increased linearly with increased loads. CONCLUSIONS: The EMG/force relationship of all quadricep muscles studied appears to be similar in isometric multi-joint LP and single-joint KE actions at a knee angle of 90 degrees. This would indicate the strategy of reciprocal force increment among muscles involved is comparable in the two models. Furthermore, these data suggest a nonuniform recruitment pattern among the three superficial QF muscles and surface EMG recordings from VL to be most reliable in predicting force output. PMID- 10694133 TI - Determination of the velocity associated with VO2max. AB - PURPOSE: The theoretical velocity associated with VO2max (vVO2max) defined by Daniels (1985) is extrapolated from the submaximal VO2-velocity relationship. VO2 is generally determined by assuming that the aerobic response reacts like a linear first-order system at the beginning of square-wave exercise with a steady state reached by the 4th minute. However, at supra-ventilatory threshold work rates, the steady state in VO2 is delayed or not attained. METHODS: The present study was carried out to compare three values for vVO2max determined with Daniels' method, but with VO2 either measured at the 4th minute (vVO2max4), the 6th minute (vVO2max6), or after the attainment of the true steady-state (vVO2maxSS). The metabolic response during square-wave exercise at each of the three vVO2max were also assessed. RESULTS: These velocities were significantly different (P < 0.05), but vVOmaxSS and vVO2max6 were highly correlated (r = 0.98; P < 0.05). Blood lactate concentrations measured after exercise at velocities very close to the three vVO2max were similar and the end-exercise VO2 were not different from VO2max, but the time required to elicit 95% VO2max during these three square-wave tests were significantly different. CONCLUSION: Therefore, when vVO2max is determined by extrapolation from the submaximal VO2-velocity relationships, submaximal VO2 should be measured beyond the 6th minute of square wave exercise (at least if it takes 30 s to reach the desired velocity) to ensure that all vVO2max reported in future studies describe a similar quantitative index. PMID- 10694134 TI - Influence of midsole bending stiffness on joint energy and jump height performance. AB - PURPOSE: A substantial amount of rotational energy is lost at the metatarsophalangeal joint during running and jumping. We hypothesized that the lost energy could be decreased by increasing the bending stiffness of shoe midsoles. The purposes of this investigation were to determine the influence of stiff shoe midsoles on changes in lower extremity joint power during running and jumping and to determine the influence of stiff shoe midsoles on vertical jump performance. METHODS: Carbon fiber plates were inserted into shoe midsoles and data were collected on five subjects during running and vertical jumping. RESULTS: The data showed that energy generation and absorption at each of the ankle, knee, and hip joints was not influenced by the stiffness of the shoe midsole. The stiff shoes with the carbon fiber plates did not increase the amount of energy stored and reused at the metatarsophalangeal joint; however, they reduced the amount of energy lost at this joint during both running and jumping. Vertical jump height was significantly higher (average, 1.7 cm for a group of 25 subjects) while wearing the stiff shoes. CONCLUSIONS: Increasing the bending stiffness of the metatarsophalangeal joint reduced the amount of energy lost at that joint and resulted in a corresponding improvement of performance. PMID- 10694135 TI - Control of maximal and submaximal vertical jumps. AB - PURPOSE: It was investigated to what extent control signals used by human subjects to perform submaximal vertical jumps are related to control signals used to perform maximal vertical jumps. METHODS: Eight subjects performed both maximal and submaximal height jumps from a static squatting position. Kinematic and kinetic data were recorded as well as electromyographic (EMG) signals from eight leg muscles. Principal component analysis was used analyze the shape of smoothed rectified EMG (SREMG) histories. Jumps were also simulated with a forward dynamic model of the musculoskeletal system, comprising four segments and six muscles. First, a maximal height jump was simulated by finding the optimal stimulation pattern, i.e., the pattern resulting in a maximum height of the mass center of the body. Subsequently, submaximal jumps were simulated by adapting the optimal stimulation pattern using strategies derived from the experimental SREMG histories. RESULTS: SREMG histories of maximal and submaximal jumps revealed only minor differences in relative timing of the muscles between maximal and submaximal jumps, but SREMG amplitude was reduced in the biarticular muscles. The shape of the SREMG recordings was not much different between the two conditions, even for the biarticular muscles. The simulated submaximal jump resembled to some extent the submaximal jumps found in the experiment, suggesting that differences in control signals as inferred from the experimental data could indeed be sufficient to get the observed behavior. CONCLUSIONS: The results fit in with theories on the existence of generalized motor programs within the central nervous system, the output of which is determined by the setting of parameters such as amplitude and relative timing of control signals. PMID- 10694136 TI - Influence of the menstrual cycle phase and menstrual symptoms on maximal anaerobic performance. AB - PURPOSE: This study was designed to analyze the effect of the menstrual cycle phase on maximal anaerobic performance during short-term anaerobic tests. METHODS: Seven eumenorrheic women (NOC) and 10 women using monophasic oral contraceptives (OC) performed three anaerobic tests (force-velocity, multi-jump, and squatting jump tests) during menstruation (M: between days 1 and 4), the midfollicular phase (F: between days 7 and 9), and the midluteal phase (L: between days 19 and 21) of the ovarian cycle. Follicular and luteal phases were confirmed by serum progesterone levels. The order of testing sessions was randomly assigned and a 15-min standardized warm-up preceded each testing session. Rectal temperatures were taken before (Trec(b)) and after (Trec(a)) warm up. RESULTS: No significant differences were observed among M, F, and L in Trec(b), Trec(a) maximal cycling power (Pmax(c)), maximal jumping power (Pmax(j)), or maximal height of jump (h(j)) in either NOC or OC. Ten of the women suffered premenstrual or menstrual symptoms (MS); the other seven did not report any premenstrual or menstrual discomfort (NMS). Presence or absence of symptoms was not correlated with oral contraceptive use. No significant differences were observed among the three stages of the menstrual cycle in Pmax(c), Pmax(j), or h(j) in NMS. In MS, only Pmax(j) decreased by 8% in M compared with that in F (P < 0.05). CONCLUSIONS: Although there were no significant differences in maximal anaerobic performance during different menstrual cycle phases, results of this study suggest that the presence or absence of premenstrual or menstrual syndrome symptoms may have an effect, possibly through an action on the stretch-shortening cycle of tendons and ligaments. PMID- 10694137 TI - Testing peak cycling performance: effects of braking force during growth. AB - The purpose of this study was to investigate the relationship between cycling peak power (CPP; flywheel inertia included) and the applied braking force (F(B)) on a friction-loaded cycle ergometer in male children, adolescents, and adults. A total of 520 male subjects aged 8-20 yr performed three brief maximal sprints against three F(B): 0.245, 0.491, and 0.736 N x kg(-1) body mass (BM) (corresponding applied loads: 25 [F(B)25], 50 [F(B)50], and 75 [F(B)75] g x kg( 1) BM). For each F(B), peak power (PP) was measured (PP25, PP50 and PP75). For each subject, the highest PP was defined as CPP. Results showed that PP was dependent on F(B). In young adults PP25 underestimated CPP by more than 10%, and consequently, F(B)25 seemed to be too low for this population. However, in children, PP75 underestimated CPP by about 20%. A F(B) of 0.736 N x kg(-1) BM was definitively too high for the pediatric population. Therefore, the optimal F(B), even corrected for BM, was lower in children than in adults. The influence of growth and maturation on the force-generating capacity of the leg muscles may explain this difference. In this study, however, it was shown that the difference between PP50 and CPP was independent of age for the whole population investigated. Consequently, when flywheel inertia is included, one cycling sprint with a F(B) of 0.495 N x kg(-1) BM (corresponding applied load: 50 g x kg(-1) BM) is a feasible method for testing both children, adolescents, or young adults. PMID- 10694138 TI - The effect of caffeine on endurance performance after nonselective beta adrenergic blockade. AB - PURPOSE: This study was designed to test the hypothesis that combined administration of propranolol and caffeine (Pr+C) would increase endurance performance compared with the administration of propranolol alone (Pr) if caffeine would be able to increase plasma free fatty acid (FFA) availability and/or lower plasma potassium concentration compared with propranolol administration alone. METHODS: Fifteen volunteers participated in the double blind placebo-controlled randomized cross-over study. An endurance exercise test until exhaustion was performed after ingestion of placebo (Pl), 80-mg propranolol (Pr), and 80-mg propranolol plus 5 mg x kg(-1) caffeine (Pr+C). RESULTS: Endurance time (+/-SD) was 79.3 +/- 20.4 min in the Pl trial, 22.6 +/- 10.8 min in the Pr trial and 31.2 +/- 17.2 min in the Pr+C trial (P < 0.001). The difference between the Pr and Pr+C trials just failed to reach statistical significance (P = 0.056). Plasma FFA concentration and plasma potassium concentrations were similar in the Pr and Pr+C trials, but differed significantly from the Pl trial (P < 0.05). CONCLUSION: Although there was a clear tendency for an improved performance in the Pr+C trial compared to the Pr trial, this improvement was not associated with increased plasma FFA concentration and/or reduced plasma potassium concentration in the Pr+C compared to the Pr trial. These results do not support the hypothesis that caffeine improves endurance performance by stimulating lipolysis or lowering plasma potassium concentration. PMID- 10694139 TI - Effect of commuter cycling on physical performance of male and female employees. AB - PURPOSE: The purpose of this study was to determine the effect of commuter cycling on physical performance. Eighty-seven male and 35 female employees volunteered to cycle regularly to their work. METHODS: Sixty-one participants went commuter cycling for 1 yr (cycling group); the others cycled only in the second half year (control group). A maximal exercise test on a cycle ergometer was carried out at the start of the study, after 6 months, and after 1 yr to measure maximal external power (Wmax) and maximal oxygen uptake (VO2max). RESULTS: After the first 6 months of commuter cycling, with a mean single trip distance of 8.5 km and a mean frequency of more than three times a week, a significant increase of 13% was found in the Wmax per kilogram body weight (Wmax x kg(-1)) in both sexes of the cycling group. The improvement in VO2max x kg(-1) was significant for the male participants (6%) but not for the female participants (-2%). At the end of the second half year, the control group also showed a mean gain in Wmax x kg(-1) of 13%. Their VO2max x kg(-1) declined in the first half year, but this was counteracted in the second half year. A dose response relationship was found between two independent variables and the physical performance; the lower the physical performance at the start of the study and the higher the total amount of kilometers cycled, the higher the gain in Wmax. For subjects with a low initial fitness level, a single trip distance of only 3 km turned out to be enough to improve physical performance. CONCLUSION: Commuter cycling can yield much the same improvement in physical performance as specific training programs. PMID- 10694140 TI - Physiological responses to a 6-d taper in middle-distance runners: influence of training intensity and volume. AB - PURPOSE: This study examined some physiological and performance responses to a 6 d taper, and the influence of training intensity and volume on these responses. METHODS: After 15 wk of training, 8 well-trained male middle-distance runners were randomly assigned to either a moderate volume taper (MVT, N = 4) or a low volume taper (LVT, N = 4), consisting of either a 50% or a 75% progressive reduction in pretaper low intensity continuous training (LICT) and high intensity interval training (HIIT). Blood samples were obtained and 800-m running performance was measured before and after taper. RESULTS: Performance was not significantly enhanced by either taper protocol (post- vs pre-taper times 124.9 +/- 4.5 vs 126.1 +/- 4.2 s with LVT, 126.2 +/- 8.0 vs 125.7 +/- 6.6 s with MVT). For the entire group of 8 subjects, red cell count, hemoglobin (Hb), mean corpuscular volume and mean corpuscular Hb concentration significantly decreased with taper, while reticulocyte count increased. Performance changes for all subjects correlated with changes in postrace peak blood lactate concentration (r = 0.87, P < 0.01). Taper LICT correlated with changes in Hb (r = 0.77), hematocrit (r = 0.81), reticulocyte count (r = 0.73), creatine kinase (r = 0.72), and total testosterone (r = -0.78), and with posttaper red cell distribution width (r = -0.75) and lymphocyte count (r = -0.82). Taper HIIT correlated nonsignificantly with changes in red cell count (r = -0.66) and total testosterone (r = 0.68). CONCLUSION: It is concluded that taper-induced physiological changes in trained middle-distance runners are mainly hematological, and that distinct physiological changes are elicited from LICT and HIIT during taper. Middle-distance runners can progressively reduce their usual training volume by at least 75% during a 6-d taper. PMID- 10694141 TI - Creatine supplementation and sprint performance in soccer players. AB - PURPOSE: This investigation examined the effects of creatine (Cr) supplementation on intermittent high-intensity exercise activities specific to competitive soccer. METHODS: On two occasions 7 d apart, 17 highly trained male soccer players performed a counter-movement jump test (CMJT), a repeated sprint test (RST) consisting of six maximal 15-m runs with a 30-s recovery, an intermittent endurance test (IET) consisting of forty 15-s bouts of high-intensity running interspersed by 10-s bouts of low-intensity running, and a recovery CMJT consisting of three jumps. After the initial testing session, players were evenly and randomly included in a CREATINE (5 g of Cr, four times per day for 6 d) or a PLACEBO group (same dosage of maltodextrins) using a double-blind research design. RESULTS: The CREATINE group's average 5-m and 15-m times during the RST were consistently faster after the intervention (0.95 +/- 0.03 vs 0.97 +/- 0.02 s, P < 0.05 and 2.29 +/- 0.08 vs 2.32 +/- 0.07 s, P = 0.07, respectively). Neither group showed significant changes in the CMJT or the IET. The CREATINE group's recovery CMJT performance relative to the resting CMJT remained unchanged postsupplementation, whereas it tended to decrease in the PLACEBO group. CONCLUSION: In conclusion, acute Cr supplementation favorably affected repeated sprint performance and limited the decay in jumping ability after the IET in highly trained soccer players. Intermittent endurance performance was not affected by Cr. PMID- 10694142 TI - Effect of mathematical modeling on the estimation of critical power. AB - PURPOSE: The purposes of this study were to re-examine the findings of previous studies by comparing the critical power (CP) estimates from five mathematical models and to determine the time to exhaustion during cycle ergometry at the lowest CP estimate from the five models. METHODS: Nine adult males performed a maximal incremental test to determine peak power and five or six randomly ordered trials on a cycle ergometer for the estimation of CP. Two linear, two nonlinear, and one exponential mathematical model were used to estimate CP. The subjects then completed two trials to exhaustion, or 60 min, at their lowest estimate of CP from the five models. RESULTS: The nonlinear three-parameter model (Nonlinear 3) produced a mean CP that was significantly (P < 0.05) less than the mean CP values derived from the other four models and was the lowest CP estimate for each subject. Two and three subjects, however, did not complete 60 min of cycling during the first and second trials at CP, respectively. At the end of the trials the subjects who completed 60 min of cycling had a mean heart rate of 92% of their maximum and a mean rating of perceived exertion of 17. CONCLUSION: These findings support previous studies that have indicated that in many cases CP overestimates the power output that can be maintained for at least 60 min. PMID- 10694143 TI - Validation of near-infrared interactance and skinfold methods for estimating body composition of American Indian women. AB - PURPOSE: This study tested the predictive accuracy of the Jackson et al. skinfold (SKF) equations (sigma7SKF and sigma3SKF), a multi-site near-infrared interactance (NIR) prediction equation, and the Futrex-5000 NMR equation in estimating body composition of American Indian women (N = 151, aged 18-60 yr). METHODS: Criterion body density (Db) was obtained from hydrodensitometry at residual lung volume. RESULTS: Sigma7SKF significantly underestimated Db (P < 0.05). Sigma3SKF and Heyward's NIR equations significantly overestimated Db (P < 0.05). The Futrex-5000 NIR equation significantly underestimated percent of body fat (%BF) (P < 0.05). Prediction errors for SKF and multi-site NIR exceeded 0.0080 g x cc(-1). The SEE for Futrex-5000 was 5.5%BF. Thus, ethnic-specific SKF and NIR equations were developed. For the SKF model, the sigma3SKF (triceps, axilla, and suprailium) and age explained 67.3% of the variance in Db:Db = 1.06198316 -0.00038496(sigma3SKF) -0.00020362(age). Cross-validation analysis yielded r = 0.88, SEE = 0.0068 g x cc(-1), E = 0.0070 g x cc(-1), and no significant difference between predicted and criterion Db. For the NIR model, the hip circumference, sigma2AdeltaOD2 (biceps and chest), FIT index, age, and height explained 73.9% of the variance in Db:Db = 1.0707606 -0.0009865(hip circumference) -0.0369861(sigma2deltaOD2) + 0.0004167(height) + 0.0000866(FIT index) -0.0001894(age). Cross-validation yielded r = 0.85, SEE = 0.0076 g x cc( 1), E = 0.0079 g x cc(-1), and a small, but significant, difference between predicted and criterion Db. CONCLUSIONS: We recommend using the ethnic-specific SKF and NIR equations developed in this study to estimate Db of American Indian women. PMID- 10694144 TI - Skinfold thickness varies directly with spring coefficient and inversely with jaw pressure. AB - PURPOSE: The main aims of this study were to: 1) determine whether heavy use of Harpenden calipers caused deterioration of the spring coefficient (force per unit length), 2) to quantify the change in skinfold thickness per unit change in jaw closing (downscale) pressure, and 3) to develop a calibration range for these calipers. METHODS: Part a) The change in spring force per unit length after at least 100,000 cycles of opening and closing five different springs was measured on a load cell. Part b) The dynamic downscale jaw pressure exerted by six pairs of Harpenden springs was measured on one caliper. Two were new pairs of springs (N1 and N2), two were 25-yr-old springs (O1 and O2), and two pairs (S1 and S2) had been used for less 1 yr. The six spring pairs were used to measure skinfold thicknesses at nine sites, in triplicate, on 20 subjects with the order of springs randomized and counterbalanced. Part c) The downscale jaw pressure of 78 Harpenden calipers was measured at eight jaw gaps. RESULTS: Part a) The springs did not change their characteristics after >100,000 cycles. Part b) At each skinfold site, the lowest thickness was recorded for S2 which exerted the highest jaw pressure (9.04 g x mm(-2)) and conversely the highest thickness was for N1 which exerted the lowest jaw pressure (8.02 g x mm(-2)). Increasing the downscale jaw closing pressure from 8.0 to 9.0 g x mm(-2) reduced a skinfold thickness by approximately 10%. Part c) The mean downscale jaw pressure was 7.82 +/- 0.25 g x mm(-2). CONCLUSIONS: In summary, it is suggested that if accurate skinfold measures between different Harpenden calipers are required, the downscale jaw pressure should be in the range of 7.40-7.82 and 7.85-8.21 g x mm(-2), at jaw gaps of 5 and 40 mm, respectively. These jaw pressures can be achieved by servicing the caliper pivot and indicator gauge to minimize frictional losses, adjusting the caliper jaw alignment, and by selecting springs that have a spring coefficient in the range 1.10-1.15 N x mm(-1). PMID- 10694145 TI - Carriage of a new epidemic strain of Neisseria meningitidis and its relationship with the incidence of meningococcal disease in Galicia, Spain. AB - In Galicia, Spain, a dramatic increase in the incidence of meningococcal disease was seen in the 1995-6. The annual incidence rose to 11 per 10(5) inhabitants, and 80% of identified strains were C:2b:P1.2,5. This led to the implementation of an intensive A+C vaccination campaign for the population aged 18 months to 19 years. During this campaign the prevalence of carriage in areas with high and low incidence was studied. Nasopharyngeal swabs were taken from 9796 subjects immediately before the administration of meningococcal vaccine, plated onto Thayer-Martin plates, incubated and sent for analysis to the Reference Laboratory for Neisseria in Spain. The prevalence of the C:2b: P1.2,5 strains was 0.6% (95% CI 0.29-0.88) in the high incidence area, and 0.41% (95 % CI 0.00-1.04) in the low incidence area, and that of serogroup C (all strains) 1.36% (95% CI 0.80 1.80) and 0.89% (95% CI 0.09-1.69) respectively. The prevalence of N. meningitidis (all strains) was almost the same in both areas (8%). Carriers of the epidemic strain were not found in the 2-4 year age group, that most affected by the disease. Our data showed a wide distribution but a low carriage rate of the epidemic strain C:2b:P1.2,5 in the high and low disease incidence areas studied; the difference in the carriage rates between the two areas was not statistically significant. PMID- 10694146 TI - Predicting the course of meningococcal disease outbreaks in closed subpopulations. AB - A stochastic epidemic model was applied to meningococcal disease outbreaks in defined small populations such as military garrisons and schools. Meningococci are spread primarily by asymptomatic carriers and only a small proportion of those infected develop invasive disease. Bayesian predictions of numbers of invasive cases were developed, based on observed data using a stochastic epidemic model. We used additional data sets to model both disease probability and duration of carriage. Markov chain Monte Carlo sampling techniques were used to compute the full posterior distribution which summarized all information drawn together from multiple sources. PMID- 10694147 TI - Outbreak of meningococcal disease in western Norway due to a new serogroup C variant of the ET-5 clone: effect of vaccination and selective carriage eradication. AB - A new sulphonamide resistant (SR) C: 15:P1.7,16 meningococcal strain, a variant of the ET-5 clone, dominated in an outbreak of 22 cases in western Norway commencing in 1995. The first eight patients were 15-21 years old from the Nordhordland area, initiating a carrier study in the local high schools. Carriage of SR serogroup C meningococci was detected by routine methods and treated with a single dose of ofloxacin 400 mg. Of 20 treated carriers, 14 harboured the outbreak strain C: 15:P1.7,16. Vaccination of 4000 children, adolescents and close contacts of patients was also performed. After the intervention, 14 additional cases of meningococcal disease occurred, 8 due to the outbreak strain. However, incidence rates dropped from 180 to 30 per 100000 per year in the student population, but increased from 0 to 13 in the rest of the population in Nordhordland. Carriage eradication is not generally recommended in Norway. However, tracing and treating meningococcal carriage may have reduced transmission and disease in this outbreak situation. PMID- 10694148 TI - Nasopharyngeal colonization of infants in southern India with Streptococcus pneumoniae. AB - To investigate the dynamics of nasopharyngeal colonization with Streptococcus pneumoniae, and to determine the prevalent serogroups/types (SGT) and their antimicrobial susceptibility, we studied 100 infants attending our well-baby clinic. Nasopharyngeal swab specimens were obtained at 6, 10, 14, 18 and 22 weeks and at 9 and 18 months of age and submitted for culture, serotyping and antimicrobial susceptibility testing of S. pneumoniae. Colonization with pneumococcus was seen on at least one occasion in 81 infants. The median age of acquisition was 11 weeks and the median duration of carriage was 1 3 months. The common SGTs identified were 6, 19, 14 and 15. SGT 1, which was a common invasive isolate in children in our hospital during this period, was not isolated from these children. Sequential colonization by 2, 3 or 4 SGTs was observed in 18, 5 and 2 children, respectively. Resistance to penicillin, chloramphenicol, cotrimoxazole and erythromycin was observed in 0, 13 (6%) 11 (5 %) and 5 (3 %) isolates, respectively. There was a significant difference in susceptibility to cotrimoxazole between colonizing and invasive isolates (5 % vs. 40 %, P<0.0001). PMID- 10694149 TI - The Hib immunization programme in the Oxford region: an analysis of the impact of vaccine administration on the incidence of disease. AB - In May 1991 an immunization programme against Haemophilus influenzae type b (Hib) infection began within the Oxford region. We validate a deterministic mathematical model of Hib by comparison with the incidence of disease in the Oxford region, 1985-97. The comparison of model results with observed outcome allows an exploration of some of the poorly understood properties of the immunization programme. Model results and observed incidence are consistent with a vaccine that blocks the acquisition of carriage. Similarly, the data suggest that factors other than experience of Hib carriage are likely to have generated acquired immunity to Hib disease prior to the introduction of vaccination. Hence it is unlikely that waning of vaccine-derived protection will result in a resurgence of disease. The inclusion in the immunization schedule of a booster dose, as used in other countries, would have provided very little extra benefit. PMID- 10694150 TI - Bordetella pertussis surveillance in England and Wales: 1995-7. AB - Available data sources on disease due to Bordetella pertussis, including notifications, hospital admissions, deaths, and an enhanced laboratory-based surveillance system commenced in January 1994, were reviewed for the period 1995 7. Pertussis notifications continued their approximately 3-year cycle although at historically reduced levels. A slight seasonal increase in late summer/early autumn existed over and above a relatively constant background rate. Over time, the proportion of pertussis cases in younger, unvaccinated children, and to a lesser extent, adolescents and young adults, is increasing. There is a continuing significant and underreported mortality associated with pertussis in the very young age group. Disease due to serotype 1,2 is on the increase despite persistent high vaccination levels and this serotype causes more severe disease. The provision of preventative antibiotics prior to disease onset reduced the severity of the disease but its use remains uncommon in England and Wales. While overall levels of pertussis notifications have declined in recent times, vaccination efficacy wanes with increasing age, and pertussis remains a significant cause of mortality and severe morbidity in the very young. This could be reduced by timely booster vaccination and increased recognition of mild disease in older cases followed by early antibiotic therapy for the very young household contacts. PMID- 10694151 TI - Characterization of enteroaggregative Escherichia coli isolated from outbreaks of diarrhoeal disease in England. AB - Twenty-two strains of enteroaggregative Escherichia coli (EAggEC), isolated from four outbreaks of diarrhoeal disease in England, were examined for a range of phenotypic attributes including the ability to produce fimbriae, haemolysins and siderophores, and cell-surface properties such as surface charge and hydrophobicity. Strains of EAggEC isolated from two of these outbreaks belonged to a diverse range of serotypes and were heterogeneous in phenotype. Strains of EAggEC isolated from the other two outbreaks belonged predominantly to serotypes 086:H34 and 098:H-, respectively. Only two strains expressed fimbriae and two strains produced an 18 kDa membrane associated protein (MAP), suggesting that EAggEC express a range of adhesion mechanisms to produce the cell arrangement recognized as the 'stacked brick' formation. The possible explanation for the diversity of EAggEC serotypes is discussed. PMID- 10694152 TI - Levels of virulence are not determined by genomic lineage of Salmonella enterica serotype Enteritidis strains. AB - Mouse virulence and the ability to adhere to, and invade cultured MDCK cells were investigated in 38 phage type reference strains of Salmonella enterica serotype Enteritidis and correlated with genomic lineage. The genomic lineage of 11 of the strains was determined in the present study; one IS200 and one ribotype pattern that had not been reported previously were observed. Log c.f.u. in the spleen 10 days post intraperitoneal (i.p.) infection with 3x10(3) bacteria (logVC10) varied between 2.9 and 8.7. The reference strains of PT7 and PT23 were found to be semi rough and were of low virulence. All other strains possessed smooth LPS. Within each of the two major clonal lines, as well as among phage types outside these, both highly virulent and moderate to low virulent strains were present. While all strains of PT1, PT2 and PT8 were highly virulent, low virulent strains were detected in PT4 and PT13. The ability to adhere to, and invade MDCK cells varied between phage types (adherence between 13 and 61% of the inocula and invasion between 4 and 151% of the adherent cells). The results of the cell culture experiments did not correlate with the results of mouse virulence tests. No correlation between clonal lineage and virulence was found within S. Enteritidis. It seems most likely that some strains have lost some of the essential factors enabling this serotype to cause successful systemic infection. PMID- 10694153 TI - Electronic network for monitoring travellers' diarrhoea and detection of an outbreak caused by Salmonella enteritidis among overseas travellers. AB - The Traveller's Diarrhoea Network, by which the Infectious Disease Surveillance Center is electronically connected with two major airport quarantine stations and three infectious disease hospitals, was launched in February 1988 in Japan. The data on travellers' diarrhoea detected is reported weekly by e-mail. Two clusters of infection among travellers returning from Italy were reported by two airport quarantine stations at the end of September 1998. A total of 12 salmonella isolates from 2 clusters were examined. All were identified as Salmonella enteritidis, phage type 4 and showed identical banding patterns on pulsed-field gel electrophoresis. A case-control study showed that the scrambled eggs served at the hotel restaurant in Rome were the likely source of this outbreak. This outbreak could not have been detected promptly and investigated easily without the e-mail network. International exchange of data on travellers' diarrhoea is important for preventing and controlling food-borne illnesses infected abroad. PMID- 10694154 TI - Acute melioidosis outbreak in Western Australia. AB - A cluster of acute melioidosis cases occurred in a remote, coastal community in tropical Western Australia. Molecular typing of Burkholderia pseudomallei isolates from culture-confirmed cases and suspected environmental sources by pulsed-field gel electrophoresis (PFGE) of XbaI chromosomal DNA digests showed that a single PFGE type was responsible for five cases of acute infection in a community of around 300 during a 5 week period. This temporal and geographical clustering of acute melioidosis cases provided a unique opportunity to investigate the environmental factors contributing to this disease. B. pseudomallei isolated from a domestic tap at the home of an asymptomatic seroconverter was indistinguishable by PFGE. Possible contributing environmental factors included an unusually acid communal water supply, unrecordable chlorine levels during the probable exposure period, a nearby earth tremor, and gusting winds during the installation of new water and electricity supplies. The possible role of the potable water supply as a source of B. pseudomallei was investigated further. PMID- 10694155 TI - Risk of transmission of tuberculosis among inmates of an Australian prison. AB - In a prison in Victoria, Australia, our objectives were contact tracing of inmates and staff at risk of exposure to an identified index case; and to determine risk factors for prevalent and incident infection. Inmates and staff who were potentially exposed to the index case were screened with a Mantoux skin test and a questionnaire. Inmate movements within the prison were compared to movements of the index case. Logistic regression was used to determine risk factors for infection. The index case had smear positive, cavitating pulmonary tuberculosis (TB), which was undiagnosed for 3 months. This was the period of potential exposure. The prevalence of positive skin test reactions in 190 inmates and staff at the prison was 10%. Significant predictors of a positive skin test were being an inmate (odds ratio (OR) 15.5), older age (OR 8.3) and being born overseas (OR 10.7). Bacille Calmette Guerin (BCG) vaccination, proximity to the index case in various prison sites, duration of incarceration, number of incarcerations and number of inmates per cell were not significant. There were three recent skin test conversions from negative to positive, representing a conversion rate of 3.5%. We did not find evidence of significant transmission of TB from a single index case. The prevalence of infection in this Australian prison was lower than published rates in other countries. Better prison conditions and different demographics of prison inmates in Australia may explain these differences. PMID- 10694156 TI - The effect of timing of sample collection on the detection of measles-specific IgM in serum and oral fluid samples after primary measles vaccination. AB - This study compares the timing of the rise and decline of measles-specific IgM in serum samples and in oral fluid samples. Two hundred and eighty 9-month-old infants presenting for routine measles vaccination in Addis Ababa, Ethiopia, were enrolled. Paired serum and oral fluid samples were collected before and 1, 2, 3 or 4 weeks after measles vaccination. Samples were tested by using a modified antibody-capture enzyme immunoassay. For the 321 IgM-negative pre- and post vaccination serum samples, 317 (99 %) of their corresponding oral fluid samples were IgM-negative. Among the 130 IgM-positive serum samples, 75% of their paired oral fluid samples were IgM-positive, with the percentage rising to 87% after oral fluid samples collected > or =3.5 weeks after vaccination were excluded. Among the post-vaccination serum samples, the percent IgM-positive peaked in week 3 and declined to 79% in week 4. For post-vaccination oral fluid samples, the percent IgM-positive peaked in weeks 2 and 3, and then declined to 43% in week 4. This modified antibody-capture enzyme immunoassay appears to detect vaccine induced measles-specific IgM in the first 3 weeks after vaccination. PMID- 10694157 TI - Intestinal protozoa in HIV-infected patients in Apulia, South Italy. AB - Protozoa are important enteric pathogens in patients with human immunodeficiency virus (HIV) infection. In this study the prevalence of intestinal protozoa in 154 HIV-infected patients, with or without diarrhoea, in our region (Apulia, South Italy) was evaluated between December 1993 and February 1998. In the majority of patients CD4+ T cell count was below 200/microl. The overall prevalence of intestinal protozoa was 43/154 (27.92%). Twenty-eight (43.08%) out of 65 patients with diarrhoea and 15 (16-85%) out of 89 non-diarrhoeic patients were parasitized. In particular, in the group of 65 patients with diarrhoea the following protozoa were identified: Cryptosporidium parvum in 14 (21.54%), Blastocystis hominis in 7 (10.77%), microsporidia in 6 (9.23%), Giardia lamblia in 4 (6.15%) and Isospora belli in 1 (1.54%). Three patients were Cryptosporidium parvum-microsporidia co-infected. In patients without intestinal symptoms, prevalence was 3/89 (3.37%) for Cryptosporidium parvum, 9/89 (10.11%) for Blastocystis hominis, 1/89 (1.12%) for microsporidia and 2/89 (2.25%) for Giardia lamblia. A significant (P<0.001) correlation was observed between protozoan infection and the presence of diarrhoea. In particular, Cryptosporidium parvum and microsporidia infections were significantly (P<0.001) and P = 0.046, respectively) associated with diarrhoeal illness. Moreover, the majority of cases of cryptosporidiosis were first diagnosed in the periods of heaviest rainfall. Therefore, drinking water contamination may be a possible source of human infection in our area. PMID- 10694158 TI - Prevalence of hepatitis B, D and C virus infections among children and pregnant women in Moldova: additional evidence supporting the need for routine hepatitis B vaccination of infants. AB - Rates of acute hepatitis B are high in Moldova, but the prevalence of chronic infection is unknown. In 1994, we surveyed children and pregnant women, collected demographic information, and drew blood for laboratory testing. Among the 439 children (mean age, 5 years), the prevalence of antibody to hepatitis B core antigen (anti-HBc) and hepatitis B surface antigen (HBsAg) were 17.1 and 6.8%, respectively. Among the 1098 pregnant women (mean age, 26 years), 52.4% were anti HBc-positive and 9.7% were HBsAg-positive. Of the HBsAg-positive pregnant women, 35.6% were hepatitis B e antigen (HBeAg) positive and 18.3% had antibodies to hepatitis D virus. The prevalence of antibody to hepatitis C virus was 1.4% in children and 2.3% in pregnant women. The high HBeAg prevalence among HBsAg positive pregnant women and the high anti-HBc prevalence among children indicate that both perinatal and early childhood transmission contribute to the high hepatitis B virus endemicity in Moldova. PMID- 10694159 TI - An outbreak of calicivirus associated with consumption of frozen raspberries. AB - In April 1988, an outbreak of gastroenteritis occurred among employees in a large company in Helsinki, Finland. A retrospective cohort study, using a self administered questionnaire, was carried out to ascertain the cause and extent of the outbreak. To meet the case definition, employees had to have had diarrhoea and/or vomiting since 2 April, 1998. A subanalysis was made in the biggest office, consisting of 360 employees, of whom 204 (57%) completed the questionnaire. Of these 108 (53%) met the case definition. Employees who had eaten raspberry dressing were more likely to meet the case definition than those who had not (Attack Rate (AR) 65% versus AR 18% Relative Risk, (RR) 3.7, 95%, Confidence Intervals (CI) 2.0-6.7). Four stool specimens obtained from affected kitchen staff who had all eaten the raspberry dressing and who had all become ill simultaneously with the employees were positive by polymerase chain reaction (PCR) for calicivirus. The data suggest that the primary source of the outbreak was imported frozen raspberries contaminated by calicivirus. PMID- 10694160 TI - Transmission of calicivirus by a foodhandler in the pre-symptomatic phase of illness. AB - After a Christmas party in a restaurant, 48 (68%) of the 82 guests contracted calicivirus gastroenteritis. The epidemiological investigation showed that salad was strongly associated with the disease episode (RR = 2.43, P = 0.0005). Similar symptoms occurred among other customers who had had a meal at the same restaurant on the same evening. A foodhandler who had only prepared salad and appetizers became sick about 30 min after the end of his shift. He had been free of symptoms while preparing food. Few outbreak investigations have shown calicivirus transmission by foodhandlers some hours before becoming symptomatic. PMID- 10694161 TI - The duration of human ocular Chlamydia trachomatis infection is age dependent. AB - We studied the relationship between age and prevalence, duration and incidence of clinical and laboratory evidence of ocular Chlamydia trachomatis infection in a cohort of Gambian subjects examined bi-weekly for 6 months. The duration of disease and infection, estimated by stratified survival analysis, proportional hazards regression and Weibull modelling, was markedly age-dependent. The estimated median duration of disease was 13.2 weeks in 0-4-year-old subjects and 1.7 weeks in those age 15 and over. Adjustment for multiple infections, and for missing observations did not alter this trend. The cumulative incidence rate of disease was reduced threefold with age. More rapid disease resolution is the main source of reduction in prevalence of active trachoma and ocular C. trachomatis infection with age; disease incidence was reduced to a lesser extent. This age dependent resolution may be effected by adaptive cellular immune mechanisms. Mechanisms responsible for natural immunity should receive appropriate emphasis in vaccine design. PMID- 10694162 TI - Planning chemotherapy based schistosomiasis control: validation of a mathematical model using data on Schistosoma haematobium from Pemba, Tanzania. AB - A mathematical model, based on a deterministic differential equation framework, has been developed to predict the impact of community chemotherapy programmes for human schistosomiasis. Here, this model is validated using data collected from a long-term control programme for urinary schistosomiasis on the island of Pemba, Zanzibar, United Republic of Tanzania, initiated in 1986 and still ongoing, in which schoolchildren were offered praziquantel chemotherapy every 6 months. Prevalence of infection and blood in urine were monitored in all the schools (total 26000 children from 60 schools) and more detailed data were collected in selected evaluation schools. Model predictions were run by using the initial prevalence as input. The predictions were very close to the observed decreases in prevalence and in prevalence of blood in urine. The correspondence improved further when the data were combined, going from single school level to district, and when the entire data set was combined. The accuracy of the predictions suggests that this model could be used as a tool to predict the consequences of chemotherapy control programmes. It is currently in press as a Windows software package under the name of 'EpiSchisto'. PMID- 10694163 TI - The potential of latent class analysis in diagnostic test validation for canine Leishmania infantum infection. AB - Accuracy assessment of diagnostic tests may be seriously biased if an imperfect reference test is used such as parasitology in the diagnosis of visceral leishmaniasis. We compared classical validity analysis of serological tests for Leishmania infantum with Latent Class Analysis (LCA), to assess whether it circumvented the gold standard problem. Clinical status, three serological tests (IFAT, ELISA and DAT) and parasitological data were recorded for 151 dogs captured in an endemic area. Sensitivity and specificity estimates from the 2x2 contingency tables were broadly corroborated by LCA, but the latter method provided more precise estimates that were robust for the different fitted models. It furthermore yielded a higher prevalence of infection and indicated that parasitology was only 55% sensitive. LCA seems a promising technique for test validation, but caution is required when applying it to sparse data sets. The feasibility and applicability of LCA in infectious disease epidemiology is discussed. PMID- 10694164 TI - Seven-week interval between acquisition of a meningococcus and the onset of invasive disease. A case report. AB - Invasive meningococcal disease (IMD) is thought to occur within a few days of pharyngeal acquisition of Neisseria meningitidis. During a longitudinal study of carriage and acquisition among 2453 first-year undergraduates we identified a male student from whom N. lactamica was isolated in October 1997 followed by N. meningitidis in December 1997. In mid-January 1998 this student suffered a mild episode of IMD (meningitis) during which N. meningitidis was isolated from his CSF. The meningococcus carried in December 1997 was phenotypically and genotypically indistinguishable from the invading organism, suggesting the possibility that the organism may have been carried for 7 weeks prior to the onset of invasive disease. Further studies are needed to assess more accurately the range of asymptomatic carriage prior to disease onset. PMID- 10694165 TI - Determination of natural versus laboratory human infection with Mayaro virus by molecular analysis. AB - A laboratory worker developed clinical signs of infection with Mayaro virus (Togaviridae), an arbovirus of South and Central America, 6 days after preparation of Mayaro viral antigen and 10 days after a trip to a rain forest. There was no evidence of skin lesions during the antigen preparation, and level 3 containment safety measures were followed. Therefore, molecular characterization of the virus was undertaken to identify the source of infection. RT-PCR and DNA sequence comparisons proved the infection was with the laboratory strain. Airborne Mayaro virus contamination is thus a hazard to laboratory personnel. PMID- 10694166 TI - Household members of hepatitis C virus-infected people in Hafizabad, Pakistan: infection by injections from health care providers. AB - Household members of people with hepatitis C are at increased risk of HCV infection. The prevalence and routes of transmission of HCV to household members in Hafizabad, Pakistan were investigated. Household members of 24 index cases were given a risk factor questionnaire, tested for HCV infection, and the risk factors between the infected and uninfected were compared. Twelve of 74 household members (16.2%) were seropositive for HCV antibody. This was 2(1/2) times the rate of infection in the general population (OR = 2.8; P = 0.01). None of the routes of transmission studied within the household was associated with an increased risk. Household members who received more than 4 injections per year were 11.9 times more likely to be infected than those who had not (P = 0.016). In Hafizabad, the greatest risk for HCV infection to household members of infected people is injections given by health-care workers rather than household contact with infected persons. PMID- 10694167 TI - Electricity-associated injuries II: outdoor management of lightning-induced casualties. PMID- 10694168 TI - Paramedic activities, drug administration and survival from out of hospital cardiac arrest. AB - OBJECTIVE: To examine the impact of administration of cardioactive drugs on the outcome from out of hospital cardiac arrest. DESIGN: Longitudinal observational cohort study with historical controls before and after the introduction of drug use in cardiac arrest by paramedics. SUBJECTS: Adult patients who had sustained an out of hospital cardiac arrest of cardiac aetiology and were treated by paramedics. SETTING: Edinburgh, Scotland. OUTCOME MEASURES: Return of spontaneous circulation, admission to and discharge from hospital. RESULTS: There was no significant difference in the demographics between Period 1 (prior to drug administration) and Period 2 (after). There was no difference in outcome between Period 1 and Period 2 for all three parameters, return of spontaneous output 30.1 versus 35%, admission to hospital 18.9 versus 24.5% and discharge 5.8 versus 6.5%. If the presenting rhythm of VF/pulseless VT alone was considered survival to hospital discharge was 12.1% in Period 1 and 10.3% in Period 2. CONCLUSION: The addition of cardioactive drug administration to the treatment of out of hospital cardiac arrest does not improve survival. PMID- 10694169 TI - Cardiopulmonary resuscitation performance of subjects over forty is better following half-hour video self-instruction compared to traditional four-hour classroom training. AB - Cardiopulmonary resuscitation (CPR) training is not well targeted to family members of individuals at highest risk of cardiac arrest. Participants in traditional CPR classes (TRAD) average 31 years of age, while family members of cardiac patients average 55 years. Video self-instruction (VSI) can reach older individuals and others who do not participate in TRAD classes. VSI is a combination of a 34-min videotape and an inexpensive manikin intended for use in the home, where three-quarters of all out-of-hospital cardiac arrests occur. We exposed 202 subjects 40 years of age and older (mean age 59.4 years, S.D. = 10.9) to either TRAD or VSI, and tested them individually immediately following training rising validated methods including measurement by means of a Laerdal Skillmeter manikin. According to American Heart Association (AHA) criteria, VSI subjects performed an average of 20.8% of all compressions and 25.1% of all ventilations correctly, compared with 3.4% of compressions and 1.7% of ventilations by TRAD subjects (P < 0.0001). VSI subjects performed an average of 10.1 of the total 14 CPR assessment and sequence skills correctly, compared with an average of 4.7 for TRAD (P < 0.0001). On a measure of overall performance, 62.7% of the VSI subjects were rated 'competent' or better (i.e. capable of performing CPR that 'would probably be effective'), compared to 6.1% of TRAD subjects (P < 0.0001). Only 17.8% of VSI subjects were rated as 'not competent' (i.e. unable to obtain a combination of any chest rise and any compression of the sternum) compared with 69.1% of TRAD subjects. VSI provides an effective, convenient, and inexpensive means of training persons over 40 years of age that achieves skill performance superior to TRAD. PMID- 10694170 TI - A scoring system for the assessment of basic life support ability. AB - INTRODUCTION: There are no current validated scoring systems for the assessment of adult single rescuer basic life support (BLS) ability. A system was proposed and prospectively validated. METHODS: The system was tested firstly on 12 skilled BLS providers (all instructors). It was then further evaluated on 75 undergraduate dental students, who were assessed before and after a standard training session on adult BLS. RESULTS: All 12 skilled persons passed the test according to the system. The system successfully showed a positive training effect in the dental students. It correctly identified those who 'passed' after training, i.e. those were capable of providing effective BLS (71/75, 94.7%). It also correctly identified those who were not considered competent (4/71, 5.3%). CONCLUSION: This is a simple, effective, objective system for assessment of basic life support. It is easily adaptable for the 1998 Guidelines on BLS. PMID- 10694171 TI - An analysis of the efficacy of bag-valve-mask ventilation and chest compression during different compression-ventilation ratios in manikin-simulated paediatric resuscitation. AB - The ideal chest compression and ventilation ratio for children during performance of cardiopulmonary resuscitation (CPR) has not been determined. The efficacy of chest compression and ventilation during compression ventilation ratios of 5:1, 10:2 and 15:2 was examined. Eighteen nurses, working in pairs, were instructed to provide chest compression and bag-valve-mask ventilation for 1 min with each ratio in random on a child-sized manikin. The subjects had been previously taught paediatric CPR within the last 3 or 5 months. The efficacy of ventilation was assessed by measurement of the expired tidal volume and the number of breaths provided. The rate of chest compression was guided by a metronome set at 100/min. The efficacy of chest compressions was assessed by measurement of the rate and depth of compression. There was no significant difference in the mean tidal volume or the percentage of effective chest compressions delivered for each compression-ventilation ratio. The number of breaths delivered was greatest with the ratio of 5:1. The percentage of effective chest compressions was equal with all three methods but the number of effective chest compressions was greatest with a ratio of 5:1. This study supports the use of a compression-ventilation ratio of 5:1 during two-rescuer paediatric cardiopulmonary resuscitation. PMID- 10694172 TI - A novel wavelet transform based analysis reveals hidden structure in ventricular fibrillation. AB - We report a new method of interrogating the surface ECG signal using techniques developed in the field of wavelet transform analysis. Previously unreported structure within the ECG during ventricular fibrillation (VF) is found using a high-resolution decomposition of the signal employing the continuous wavelet transform. We believe that wavelet transform methods could lead to the development of powerful tools for use in the resuscitation of patients with cardiac arrest. PMID- 10694173 TI - The effects of epinephrine/norepinephrine on end-tidal carbon dioxide concentration, coronary perfusion pressure and pulmonary arterial blood flow during cardiopulmonary resuscitation. AB - End-tidal CO2 concentration correlates with pulmonary blood flow during cardiopulmonary resuscitation and has been claimed to be a useful tool to judge the effectiveness of chest compression. A high concentration of end-tidal CO2 has been related to a better outcome. However, most authors have noticed a decrease in end-tidal CO2 concentration after administration of epinephrine, concomitant with an increase in coronary perfusion pressure and an increased incidence of return of spontaneous circulation. This study was performed to evaluate changes in end-tidal CO2 concentration after injection of vasopressors during cardiopulmonary resuscitation and to investigate the time-course of the response and possible explanations for it. After 1 min of electrically induced cardiac arrest and 5 min of chest compressions, 18 pigs were randomly assigned to receive 0.045 mg kg(-1) epinephrine, 0.045 mg kg(-1) norepinephrine or no drug. After another 4 min of chest compressions the pigs were defibrillated. End-tidal CO2, pulmonary blood flow and coronary perfusion pressure decreased immediately after the induction of cardiac arrest, increased slightly during chest compressions and increased initially to supernormal levels after the return of spontaneous circulation. Injection of epinephrine or norepinephrine during chest compressions decreased end-tidal CO2 51 +/- 2%, (mean +/- S.E.M.), and 43 +/- 1%, respectively, and pulmonary blood flow by 134 +/- 13 and 125 +/- 16%, respectively, within 1 min, simultaneously increasing coronary perfusion pressure from 10 +/- 2 to 45 +/- 5 mm Hg and from 11 +/- 1 to 38 +/- 5 mm Hg, respectively. The coronary perfusion pressure slowly fell, but the effects on end tidal CO2 and pulmonary blood flow were prolonged. In conclusion, vasopressors increased coronary perfusion pressure and the likelihood of a return of spontaneous circulation, but decreased end-tidal CO2 concentration and induced a critical deterioration in cardiac output and thus oxygen delivery in this model of cardiopulmonary resuscitation. PMID- 10694174 TI - Amniotic fluid embolism: a case report and review. AB - A 41-year old primigravida underwent caesarean section because of foetal distress following prostin induction of labour. Intraoperative coagulopathy, haemorrhage and hypotension necessitated a hysterectomy. Subsequently, she developed respiratory and renal failure, requiring mechanical ventilation and haemodialysis. She made a full recovery. The likely diagnosis was amniotic fluid embolism (AFE), a rare complication of pregnancy with a variable presentation, ranging from cardiac arrest and death through to mild degrees of organ system dysfunction with or without coagulopathy. The differential diagnosis includes pre eclamptic toxaemia/pregnancy-induced hypertension, anaphylaxis and pulmonary embolism. There is no diagnostic test for AFE; the finding of foetal elements in the maternal circulation is non-specific. Historically, AFE was thought to induce cardiovascular collapse by mechanical obstruction of the pulmonary circulation. It is now thought that a combination of left ventricular dysfunction and acute lung injury occur, with activation of several of the clotting factors. An immunological basis for these effects is postulated. There is no specific therapy and treatment is supportive. The mortality of the condition remains high. PMID- 10694175 TI - Tracheal injury as a sequence of multiple attempts of endotracheal intubation in the course of a preclinical cardiopulmonary resuscitation. AB - Management of the difficult airway requires an appropriate approach based on personal clinical experiences. For every physician involved in rescue and emergency medicine, it is important to know the difficult airway algorithm and be familiar with alternative techniques of managing the difficult airway. We report a case of tracheal injury caused by multiple attempts at intubating the trachea. Based on current knowledge, apart from surgical equipment for cricothyroidotomy the laryngeal mask airway (LMA) and the Combitube (ETC) should be available on any ambulance vehicle staffed by an emergency physician. In future, blind intubation through the intubating laryngeal mask airway (ILMA) could offer a new opportunity. PMID- 10694176 TI - Cardiac rupture by penetration of fractured sternum: a rare complication of cardiopulmonary resuscitation. AB - We report an 82-year-old man in whom cardiopulmonary resuscitation (CPR) was unsuccessful. The postmortem examination revealed right atrial ruptures and pericardial sac perforation by a fractured sternal edge. Even though CPR-related cardiac rupture is rare, emergency medical staff should be aware of this complication. PMID- 10694177 TI - Automated defibrillation performed by emergency medical technicians: the Madrid experience. PMID- 10694178 TI - Gel pads should not be used for monitoring ECG after defibrillation. PMID- 10694179 TI - An intact renin-angiotensin system is a prerequisite for normal renal development. AB - All components of the renin-angiotensin system (RAS) are highly expressed in the developing kidney in a pattern that suggests a role for angiotensin II in renal development In support of this notion, pharmacological interruption of angiotensin II type-1 (AT1) receptor-mediated effects in animals with an ongoing nephrogenesis produces specific renal abnormalities characterized by papillary atrophy, abnormal wall thickening of intrarenal arterioles, tubular atrophy associated with expansion of the interstitium, and a marked impairment in urinary concentrating ability. Similar changes in renal morphology and function also develop in mice with targeted inactivation of the genes that encode angiotensinogen, angiotensin converting enzyme, or both AT1 receptor isoforms simultaneously. Taken together, these results clearly indicate that an intact signalling through AT1 receptors is a prerequisite for normal renal development In a recent study, an increased incidence of congenital anomalies of the kidney and urinary tract was detected in mice deficient in the angiotensin II type-2 receptor, suggesting that this receptor subtype is also involved in the development of the genitourinary tract The present report mainly reviews the renal abnormalities that have been induced by blocking the RAS pharmacologically or by gene targeting in experimental animal models. In addition, pathogenetic mechanisms and clinical implications are discussed. PMID- 10694180 TI - Relationship between blood pressure level and mortality rate: an 18-year study conducted in two rural communities in Japan. AB - BACKGROUND: There have been very few studies on life prognosis of cardiovascular disease according to blood pressure stratification in the Japanese. Therefore, in Japan, although treatment of hypertension is possible, albeit at times difficult, due to the availability of various antihypertensive medications, the appropriate time at which treatment should be started remains a problem. OBJECTIVE: To investigate the long-term prognosis of cardiovascular disease in the Japanese general population according to blood pressure stratification in the Japanese. DESIGN: A community-based prospective cohort study of 1996 men and women between the ages of 40 and 64 years at the baseline examination was conducted over an 18 year period. Information on death was obtained from local public health nurses and death certificates. The causes of death were clarified by questionnaires sent to doctors in the hospital in which the deceased was hospitalized, and the causes of death were analysed in each blood pressure category at baseline examination. RESULTS: Mortality from cardiovascular disease increased with increases in the level of blood pressure and was significantly higher in > or = 140 mmHg group in systolic blood pressure and > or = 90 mmHg group in diastolic blood pressure, adjusted for age, sex and other cardiovascular risk factors in the Japanese general population. However, mortality risk from cardiovascular disease did not have a J-shaped relationship with systolic and diastolic blood pressure. CONCLUSION: We conclude that the optimum time for starting treatment in Japanese people is when blood pressure is 140/90 mmHg or less. PMID- 10694181 TI - Hypertension prevalence and care in an urban and rural area of Tanzania. AB - OBJECTIVE: To describe the prevalence, detection, treatment and control of hypertension in an urban and rural area of Tanzania. DESIGN: Two linked cross sectional population-based surveys. SETTING: A middle-income urban district of Dar es Salaam (Ilala) and a village in the relatively prosperous rural area of Kilimanjaro (Shari). PARTICIPANTS: Seven hundred and seventy adults (> 15 years) in Ilala and 928 adults in Shari were studied. RESULTS: Hypertension prevalence (blood pressure > or = 140 and/or 90 mmHg, or known hypertensives receiving anti hypertensive treatment) was 30% (95% confidence interval, 25.1-34.9%) in men and 28.6% (24.3-32.9%) in women in Ilala, and 32.2% (27.7-36.7%) in men and 31.5% (27.8-35.2%) in women in Shari. Age-standardized hypertension (to the New World Population) prevalence was 37.3% (32.2-42.5%) among men and 39.1% (34.2-44.0%) in women in Ilala, and 26.3% (22.4-30.4%) in men and 27.4% (24A-30.4%) in women in Shari. In both areas, just under 20% of hypertensive subjects were aware of their diagnosis, approximately 10% reported receiving treatment and less than 1% were controlled (blood pressure < 140/90 mmHg). Hypertensive subjects were older, had greater body mass indices and waist: hip ratios, and had more risk factors for hypertension and its complications (smoking, heavy alcohol consumption, physical inactivity, obesity and diabetes) than non-hypertensives. CONCLUSIONS: There is a high prevalence of hypertension in rural and urban areas of Tanzania, with low levels of detection, treatment and control. This demonstrates the need for cost effective strategies for primary prevention, detection and treatment of hypertension and the growing public health challenge of non-communicable diseases in Sub-Saharan Africa. PMID- 10694182 TI - Defensiveness status predicts 3-year incidence of hypertension. AB - OBJECTIVE: A growing body of research indicates that defensive personality styles (in particular, self-deception) may be related to higher resting blood pressure and stress reactivity levels. This study is the first, however, to examine the value of defensiveness as a prognostic indicator for the development of clinical hypertension. METHODS: Participants were 127 initially normotensive male and female adults who completed a comprehensive protocol including psychological testing, assessment of smoking, physical activity and body fat levels, and 8-12 h ambulatory blood pressure monitoring. Participants returned 3-years later for an identical follow-up protocol. Defensiveness was assessed using the Balanced Inventory of Desirable Responding. RESULTS: At 3-year testing, 15 of 127 participants (12%) met criteria for hypertension (i.e. ambulatory mean blood pressure > 140/90). Comparisons between defensiveness groups showed that 12 of 60 (20%) high defensiveness participants met hypertension criteria, whereas only three of 67 (4.5%) low defensiveness participants were hypertensive. Logistic regression equations adjusted for age, alcohol usage, bodyfat, self-reported exercise levels, smoking, and year-1 ambulatory blood pressure, revealed that membership in the high defensiveness group was associated with more than a sevenfold risk of 3-year hypertension (adjusted risk ratio, 7.5; 95% confidence interval, 1.5-39.2). CONCLUSIONS: These findings link defensive characteristics to an increased prospective risk of hypertension using state of the art ambulatory monitoring techniques, and were robust after controlling for established risk factors. We conclude that the current results add to the hypertension literature by demonstrating associations between personality and clinically relevant blood pressure criteria. PMID- 10694183 TI - Analysis of the genetic basis of the endothelium-dependent impaired vasorelaxation in the stroke-prone spontaneously hypertensive rat: a candidate gene approach. AB - OBJECTIVE: To investigate the role of potential candidate genes in the pathogenesis of the endothelium-dependent impaired vasorelaxation that associates and co-segregates with stroke in the stroke-prone spontaneously hypertensive rat (SHRsp) compared with the stroke-resistant SHR (SHRsr). DESIGN AND METHODS: An SHRsp/SHRsr F2-intercross (n = 137; 64 males, 73 females) was obtained and, at the age of 6 weeks, it was placed under a stroke permissive Japanese-style diet for 4 weeks. At the end of the treatment the vascular function of each rat was characterized. The maximal vasorelaxation to acetylcholine after maximal vasoconstriction (delta ratio) was considered as the quantitative phenotype. The following candidate genes were related to the delta ratio: renin, angiotensinogen, angiotensin-converting enzyme, angiotensin II AT1b receptor, atrial natriuretic peptide, brain natriuretic peptide, atrial natriuretic peptide GC-A receptor, kallikrein, endothelial nitric oxide synthase. In addition, polymorphic markers located inside areas of the rat genome where other candidates (i.e. adrenomedullin, endothelin, Ang II AT1a receptor) are known to map were included. RESULTS: The endothelial vascular dysfunction of the SHRsp showed a variable distribution among SHRsp/SHRsr F2 descendants, independently from the blood pressure levels. A genotype/phenotype co-segregation analysis for each of the genes tested did not show any statistically significant co-segregation with the vascular phenotype. CONCLUSION: A candidate gene approach used to investigate the genetic basis of the endothelial-dependent vascular dysfunction of the SHRsp strain did not reveal any evidence to support the hypothesis that the genes tested play any role in the pathogenesis of the stroke-related vascular abnormality. PMID- 10694184 TI - Association analysis of beta2 adrenoceptor polymorphisms with hypertension in a Black African population. AB - OBJECTIVE: To determine whether or not beta2 adrenoceptor polymorphism is a risk factor for the development of hypertension in a Black South African population. BACKGROUND: Attenuated vasodilator responses to endogenous catecholamines may contribute to the aetiology of hypertension. Downregulation of beta2 adrenoreceptors (beta2AR) following stimulation with agonists is determined in part by variation at the beta2AR gene locus. The Glu27 beta2AR genotype results in attenuated downregulation compared with the wild-type Gln27 receptor, whereas Gly16 exhibits enhanced down-regulation compared to Arg16. Possible racial differences in the prevalence of the beta2AR polymorphisms may be an explanation for the blunted responses to isoprenaline and the increased prevalence of hypertension in Black African populations. METHODS: One hundred and ninety-two unrelated hypertensives and 123 normotensives of Black South African origin were studied. Hypertensives were recruited from hospital hypertension clinics in the province of Gauteng and if on treatment, had a 2-4 week washout period before 24 h ambulatory blood pressure assessment Normotensive controls were recruited from the same community. RESULTS: There was no significant association between either the Arg-Gly16 polymorphism or the Gln-Glu27 polymorphism and hypertension status. Furthermore, in the hypertensives, no significant association was seen between beta2AR genotype at either site and clinical blood pressure, 24-h blood pressure or left ventricular mass. A significant association was seen between Arg16 homozygotes and lower body mass index in hypertensives (P = 0.007) although this was not a primary end point. Interestingly, the Glu27 polymorphism was much rarer in this population (allelic frequency 17%) compared to a Caucasian population. CONCLUSION: These data suggest that beta2AR polymorphism is not a risk factor for hypertension per se in this defined population. The possibility that the decreased prevalence of Glu27 in black South African populations explains blunted vasodilator responses to isoprenaline requires further study. PMID- 10694185 TI - The L10F mutation of angiotensinogen is rare in pre-eclampsia. AB - BACKGROUND: A mutation in the gene for angiotensinogen, changing the leucine residue at position 10 to a phenylalanine (L10F), has been reported in a patient with proteinuric pre-eclampsia. In vitro enzymatic studies suggest this mutation would increase production of the vasoactive peptide, angiotensin II in vivo, and therefore explain the etiology of the maternal hypertension. OBJECTIVE: To determine whether mutation of codon 10 of angiotensinogen is common in pre eclampsia, and therefore likely to be involved in disease susceptibility. DESIGN: We collected a cohort of 32 women with 'true' pre-eclampsia. All were normotensive prior to the 20th week of pregnancy, developed blood pressures consistently above 140/90 mmHg and had proteinuria of greater than 300 mg/day during the third trimester. All had blood pressures that returned to normal within 1 month of delivery; 31 women were primigravida. Genomic DNA was isolated from their peripheral blood lymphocytes for genetic analyses. METHODS: A polymerase chain reaction-restriction enzyme-based assay was devised to screen for mutation of codon 10 of the angiotensinogen gene. In addition, we determined the frequency of a threonine residue at position 235 in the angiotensinogen gene, given previous controversial findings of association of this polymorphism with disease. CONCLUSIONS: We detected no mutation of codon 10 in angiotensinogen in any of the 32 women studied, indicating that this mutation is not commonly associated with proteinuric pre-eclampsia. Furthermore, there was no increased frequency of threonine 235 in the affected individuals studied compared with respective normotensive Caucasian-American and African-American populations. PMID- 10694186 TI - Acute production of vascular superoxide by angiotensin II but not by catecholamines. AB - OBJECTIVE: To determine whether vascular superoxide is rapidly released by angiotensin II and is involved in vascular contraction. DESIGN: The effect of superoxide dismutase (SOD) on angiotensin II induced elevation of mean arterial blood pressure was measured. Subsequently, acute production of vascular superoxide by angiotensin II and its effect on isometric tension were measured in rat aortic rings. The effects of catecholamines were concomitantly measured. METHODS AND RESULTS: The acute pressor effects of angiotensin II were significantly reduced when rats were pretreated intravenously with SOD. When angiotensin II was added on aortic segments in the presence of Cypridina luciferin analog, immediate elevations of chemiluminescence were observed which were inhibited by SOD. Furthermore, angiotensin II-induced elevations of isometric tension in aortic rings were significantly reduced by SOD. The effects of epinephrine and norepinephrine were concomitantly measured and were not significant CONCLUSIONS: The acute superoxide producing effect is likely to be specific to angiotensin II, because such a significant modification of the effects was not observed for catecholamines. Our results suggest that angiotensin II causes acute vascular superoxide production, which may be involved in the acute pressor effects. PMID- 10694187 TI - Effect of antioxidant treatments on nitrate tolerance development in normotensive and hypertensive rats. AB - OBJECTIVES: To investigate the effect of chronic antioxidant treatments on the development of nitrate tolerance in spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats by evaluating (i) coronary vascular reactivity, (ii) lipid peroxidation (malondialdehyde), and (iii) peroxynitrite formation (3-nitrotyrosine). METHODS: Tolerance was induced in 16-week-old male SHR and WKY, by 4 days of continuous treatment with nitroglycerin patches. Two groups were orally pre-treated (2-weeks) with antioxidants: N-acetyl-L-cysteine (NAC) or melatonin. Effects of serotonin (5-HT) and sodium nitroprusside (SNP) perfusion were tested in isolated Langendorff-perfused hearts. 3-Nitrotyrosine levels were measured in coronary sinus effluent and malondialdehyde in plasma. RESULTS: Nitrate tolerance reduced SNP-induced dilation in both strains. This alteration was differently improved by antioxidants: melatonin was effective in SHR, whereas NAC was effective in WKY. Tolerance also reduced 5-HT-mediated vasodilation in WKY, which was reversed by both antioxidants. By contrast, nitrate tolerance enhanced the vasoconstriction to 5-HT in SHR and both antioxidants prevented this response. Furthermore, tolerance was associated with higher malondialdehyde levels in both strains and with higher 3-nitrotyrosine levels in SHR. These changes were reversed by both antioxidants. CONCLUSIONS: A participation of oxidative stress was suggested during nitrate tolerance development, since antioxidants prevented the increase in lipid peroxidation and improved vascular responses to SNP and 5HT. Differential effects of antioxidants on SNP-induced vasodilation in SHR and WKY may suggest distinct mechanisms of tolerance development in hearts from hypertensive and normotensive rats. An increased peroxynitrite generation, expressed by higher 3-nitrotyrosine levels, could contribute to nitrate tolerance in the coronary circulation of SHR. PMID- 10694188 TI - Baseline reproducibility of B-mode ultrasonic measurement of carotid artery intima-media thickness: the European Lacidipine Study on Atherosclerosis (ELSA). AB - BACKGROUND AND OBJECTIVE: The European Lacidipine Study of Atherosclerosis (ELSA) is a prospective, randomized, double-blind, multi-national interventional trial to determine the effect of four-year treatment using the calcium antagonist lacidipine versus the beta-blocker atenolol on the progression of carotid atherosclerosis in 2259 asymptomatic hypertensive patients. B-mode ultrasound is used to measure the primary and secondary endpoints including the mean maximum intima-media thickness (IMT) of the carotid bifurcations and the common carotid arteries (CBM(max)), the mean maximum IMT of 12 standard carotid sites (M(max)) and the overall maximum IMT (T(max)). This paper reports the cross-sectional reproducibility of ultrasound measurements at baseline. METHOD: To evaluate measurement reliability, each patient is scanned twice at baseline and again at four annual visits, with 80% of the replicate scans performed by the same sonographer and 20% by a different sonographer; 50% of the replicate scans are read by the same reader and the other 50% by different readers. RESULTS: The overall coefficient of reliability (R) was 0.859 for CBM(max), 0.872 for M(max) and 0.794 for T(max). The reliability for CBM(max) was stable during the 1 3/4 year baseline period (R = 0.848 to 0.953) and was uniform among the 23 field centres (R = 0.798 to 0.926). Intra- and inter-reader reliability were 0.915 and 0.872 respectively, and intra-sonographer reliability was 0.866. CONCLUSION: The results demonstrate that by implementing standardized protocols and strict quality control procedures, highly reliable ultrasonic measurements of carotid artery IMT can be achieved in large multi-national trials. PMID- 10694189 TI - Evidence for a sympatholytic effect of mibefradil in the pithed rat preparation. AB - OBJECTIVE: The T-type prevalent calcium channel blocker mibefradil (MIB) was shown to possess N-type calcium channel blocking properties. As this particular type of calcium channel is known to be crucially involved in the neuronal release of noradrenaline, we have investigated whether MIB could be a sympatholytic drug. METHODS: To evaluate the sympathoinhibitory action, the effects of 3 and 10 micromol/kg MIB on the tachycardic effect of electrical stimulation of the preganglionic cardioaccelerator nerves in the pithed rat were investigated. The effect of MIB on the dose-response curve of externally applied noradrenaline was also studied. To compare the results with a classic L-type calcium channel blocker, the experiments were repeated with 3 and 10 micromol/kg verapamil (VER). RESULTS: The maximal increase in heart rate in response to electrical nerve stimulation was 96 +/- 7 bpm (control, n = 6), 70 +/- 6 bpm (3 micromol/kg MIB, n = 8), 57 +/- 6 bpm (10 micromol/kg MIB, n = 5), 93 +/- 5 bpm (3 micromol/kg VER, n = 6) and 46 +/- 7 bpm (10 micromol/kg VER, n = 5). The tachycardic response to electrical stimulation at 1, 5 and 10 Hz was completely blocked by 5 mg/kg intravenous guanethidine. The maximal increase in heart rate in response to noradrenaline was 96 +/- 4 bpm (control, n = 6), 103 +/- 6 (3 micromol/kg MIB, n = 6), 42 +/- 9 bpm (10 micromol/kg MIB, n = 5), 73 +/- 5 bpm (3 micromol/kg VER, n = 5) and 40 +/- 7 bpm (10 micromol/kg VER, n = 6). Under control conditions and in the presence of 3 micromol/kg MIB and VER the maximal effect of noradrenaline was reached at 0.1 micromol/kg whereas in the presence of 10 micromol/kg MIB and VER it was reached at a dose of 1 micromol/kg. MIB at a dose of 3 micromol/kg was significantly more effective in reducing the chronotropic response to electrical stimulation compared with externally applied noradrenaline. For VER the opposite holds true. These differences were not observed with doses of 10 micromol/kg MIB and VER. CONCLUSION: Mibefradil, besides its direct effect on cardiac T- and L type calcium channels, reduces the release of noradrenaline from sympathetic nerve endings, most probably by inhibition of presynaptic N-type calcium channels. In the model used this effect is only observable at relatively low concentrations, most probably because of the direct cardiodepressant action of MIB provoked by L-type channel blockade. PMID- 10694190 TI - Cicletanine reverses vasoconstriction induced by the endogenous sodium pump ligand, marinobufagenin, via a protein kinase C dependent mechanism. AB - RATIONALE: Cicletanine (CIC), an anti-hypertensive compound with direct vascular and natriuretic actions, is especially effective in salt-sensitive hypertension, in which dysregulation of the sodium pump plays an important pathogenic role, and digitalis-like cardiotonic steroids contribute to increased vascular tone. The purpose of the present study was to investigate whether, and by what mechanisms, cicletanine antagonizes the vasoconstrictor effects of cardiotonic steroids in isolated human arteries. METHODS: The effects of cicletanine on vascular tone were studied in isolated, endothelium-denuded rings of 2nd-3rd-order branches of human mesenteric arteries pre-contracted with bufodienolide marinobufagenin (MBG), an Na/K-ATPase inhibitor, or endothelin-1 (ET-1). Na/K-ATPase activity was measured in sarcolemmal membranes from the mesenteric artery. Activity of rat brain protein kinase C (PKC) was measured using the PepTag phosphorylation assay. RESULTS: MBG and ET-1 both induced sustained vasoconstriction in human mesenteric artery rings, and cicletanine relaxed rings pre-contracted with either MBG (EC50 = 11 +/- 2 micromol/l) or ET-1 (EC50 = 6.4 +/- 1.1 micromol/l). Although 8-Br cGMP (100 micromol/l) caused complete vasorelaxation of arterial rings pre contracted with ET-1, it did not affect the MBG-induced vasoconstriction. An activator of PKC, phorbol diacetate (PDA) (50 nmol/l), attenuated CIC-induced vasorelaxation of mesenteric artery rings pre-contracted with MBG (EC50 > 100 micromol/l), but not rings pre-contracted with ET-1 (EC50 = 6.5 +/- 1.2 micromol/l). In mesenteric artery sarcolemma, 100 nmol/l MBG inhibited the Na/K ATPase by 68 +/- 5% and cicletanine (100 micromol/l) attenuated this Na/K-ATPase inhibition by 85 +/- 6%. In the PepTag PKC assay, cicletanine produced a concentration-dependent inhibition of rat brain PKC activity (IC50 45 +/- 11 micromol/l). In the presence of 50 nmol/l PDA, 100 micromol/l cicletanine did not antagonize the Na/K-ATPase inhibition by MBG, and did not inhibit the PKC from rat brain. CONCLUSIONS: Cicletanine antagonizes vasoconstriction induced by Na/K ATPase inhibition via a PKC-dependent mechanism that does not involve inhibition of cyclic GMP phosphodiesterase (cGMP-PDE). This mechanism of action may be relevant to the greater potency of cicletanine in salt-sensitive hypertension in which plasma levels of endogenous digitalis-like cardiotonic steroids are elevated. Our findings also suggest that PKC is an important factor for cardiotonic steroid-Na/K-ATPase interactions on the vascular tone, and is therefore a potential target for therapeutic intervention in hypertension. PMID- 10694191 TI - Platelet alpha2-adrenoceptor alterations in patients with essential hypertension are normalized after treatment with doxazosin but not propranolol. AB - OBJECTIVE: Marked alterations have been demonstrated to occur in the platelet alpha2-adrenoceptors of patients with essential hypertension. The purpose of this study was to determine whether antihypertensive treatment with alpha-adrenergic blocker doxazosin or beta-adrenergic blocker propranolol can affect the affinity and the density of platelet alpha2-adrenoceptors in such patients. SUBJECTS AND METHODS: In two groups of 22 previously untreated, essential hypertensive patients, the mean affinity (Kd) and density (B(max)) of platelet alpha2 adrenoceptors were studied by [3H]-UK 14304 binding assays; the first assays were performed before any medication was begun, the second were performed after treatment for up to 13 weeks with doxazosin or propranolol. A third group of 22 healthy normotensive volunteers matched by age, sex and body mass index was used as control. RESULTS: Blood pressure did not differ significantly in the two hypertensive groups, and treatment with the two drugs resulted in closely similar, normal blood pressure levels. Kd and B(max) values were significantly higher in the two hypertensive groups than in controls. After treatment with propranolol the binding parameters did not change significantly, whereas after treatment with doxazosin Kd and B(max) returned to normotensive values. CONCLUSIONS: In previously untreated, essential hypertensive patients platelet alpha2-adrenoceptors have a lower affinity but a higher density than in normotensive subjects. Despite similar effects on blood pressure, the treatment with the alpha-adrenergic blocker doxazosin is followed by restoration of normal findings in the binding assays of platelet alpha2-adrenoceptors whereas the treatment with the beta-adrenergic blocker propranolol does not alter the Kd and B(max) values. PMID- 10694192 TI - Myocardial bradykinin B2-receptor expression at different time points after induction of myocardial infarction. AB - OBJECTIVE: To characterize the regulation of the myocardial bradykinin B2 receptor after induction of myocardial infarction (MI), we studied its expression at different time points in the left ventricle (LV), right ventricle (RV) and interventricular septum (S) of the heart. DESIGN: Male Sprague-Dawley rats were submitted to permanent occlusion of the left descending coronary artery. Six hours, 24 h or 6 days after MI induction or a sham operation, a Millar-tip catheter was placed in the LV. Left ventricular pressure (LVP) and contractility [(dP/dt)max] were measured. The LV, RV and S of all animals were isolated, and total RNA was extracted. B2-receptor expression was analysed by an RNase protection assay. In addition, Western blot analysis was used to determine protein levels of the B2 receptor in the infarcted area of the LV. RESULTS: We observed a decrease in LVP and contractility at all time points after MI in comparison with sham-operated animals. Basal B2-receptor expression was detected in the LV and RV, but not in the S of sham-operated rats. In the LV of infarcted hearts, we found a time-dependent up-regulation of the B2-receptor expression, which was increased twofold and fivefold, respectively, 6 h and 24 h after induction of MI compared with controls. This increase was maintained for at least 6 days. Similarly, we also found an up-regulation of the B2-receptor expression in the RV and S. Both reached a peak 24 h after induction of MI. The protein level of the receptor gradually increased up to day 6. CONCLUSION: We conclude that myocardial ischaemia triggers B2-receptor up-regulation in both the infarcted and non-infarcted areas of the heart. PMID- 10694193 TI - Effects of oral administration of L-arginine on renal function in patients with heart failure. AB - OBJECTIVES: Although the beneficial effects of L-arginine on systemic haemodynamics have been reported in patients with heart failure, its effect on renal function has not been examined. We evaluated the effects of oral administration of L-arginine on renal haemodynamics, sodium and water handling, and various hormonal factors in patients with chronic heart failure. SUBJECTS AND METHODS: A double-blind crossover trial was performed in 17 patients with chronic congestive heart failure (NYHA II-III, 56 +/- 12 years of age) who were randomly assigned to receive oral L-arginine (15 g/day) and placebo or placebo and arginine sequentially for 5 days each. Twenty-four hour creatinine clearance (Ccr), and 24-h urinary cyclic guanosine 5-monophosphate (GMP) excretion were determined. Saline loading was performed on day 5 of each treatment Renal blood flow, glomerular filtration rate (GFR), and urinary sodium excretion rate (UNa) were assessed before and after saline loading. RESULTS: Twenty-four hour GMP excretion (1.4 +/- 1.1 versus 0.8 +/- 0.5 micromol/day, P < 0.01) and Ccr (150 +/ 43 versus 125 +/- 42 ml/min, P < 0.05) were higher and plasma endothelin level (2.5 +/- 0.6 versus 3.1 +/- 0.8 pg/ml, P < 0.05) was lower with L-arginine treatment compared to placebo treatment In addition, the relative increase of UNa and GFR after saline loading were significantly higher in L-arginine treatment (UNa, 47 +/- 12%; GFR, 44 +/- 31%) than in placebo treatment (UNa, 34 +/- 9%; GFR, 22 +/- 29%) (P < 0.05). CONCLUSIONS: Oral administration of L-arginine has beneficial effects on glomerular filtration rate, natriuresis, and plasma endothelin level in patients with chronic congestive heart failure. PMID- 10694194 TI - Structure and the resistance amplifier in hypertension: reply to the dissenters. PMID- 10694196 TI - A(2A) adenosine receptors in human peripheral blood cells. PMID- 10694197 TI - Role of nitric oxide- and vasoactive intestinal polypeptide-containing neurones in human gastric fundus strip relaxations. AB - The morphological pattern and motor correlates of nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) innervation in the human isolated gastric fundus was explored. By using the nicotinamide adenine dinucleotide phosphate hydrogen (NADPH)-diaphorase and specific rabbit polyclonal NO-synthase (NOS) and VIP antisera, NOS- and VIP-containing varicose nerve fibres were identified throughout the muscle layer or wrapping ganglion cell bodies of the myenteric plexus. NOS-immunoreactive (IR) neural cell bodies were more abundant than those positive for VIP-IR. The majority of myenteric neurones containing VIP coexpressed NADPH-diaphorase. Electrical stimulation of fundus strips caused frequency-dependent NANC relaxations. N(G)-nitro-L-arginine (L-NOARG: 300 microM) enhanced the basal tone, abolished relaxations to 0.3 - 3 Hz (5 s) and those to 1 Hz (5 min), markedly reduced ( approximately 50%) those elicited by 10 - 50 Hz, and unmasked or potentiated excitatory cholinergic responses at frequencies > or =1 Hz. L-NOARG-resistant relaxations were virtually abolished by VIP (100 nM) desensitization at all frequencies. Relaxations to graded low mechanical distension (< or =1 g) were insensitive to tetrodotoxin (TTX: 1 microM) and L NOARG (300 microM), while those to higher distensions (2 g) were slightly inhibited by both agents to the same extent ( approximately 25%). In the human gastric fundus, NOS- and VIP immunoreactivities are colocalized in the majority of myenteric neurones. NO and VIP mediate electrically evoked relaxations: low frequency stimulation, irrespective of the duration, caused NO release only, whereas shortlasting stimulation at high frequencies induced NO and VIP release. Relaxations to graded mechanical distension were mostly due to passive viscoelastic properties, with a slight NO-mediated neurogenic component at 2 g distension. The difference between NO and VIP release suggests that in human fundus accommodation is initiated by NO. British Journal of Pharmacology (2000) 129, 12 - 20 PMID- 10694198 TI - Adverse effects of an active fragment of parathyroid hormone on rat hippocampal organotypic cultures. AB - Adverse effects of an active fragment of parathyroid hormone (PTH(1 - 34)), a blood Ca(2+) level-regulating hormone, were examined using rat hippocampal slices in organotypic culture. Exposure of cultured slice preparations to 0.1 microM PTH(1 - 34) for 60 min resulted in a gradual increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)); this effect was most obvious in the apical dendritic region of CA1 subfield. When PTH(1 - 34) at a lower concentration (1 nM) was added to the culture medium and its toxic effects examined using a propidium iodide intercalation method, significant toxicity was seen 3 days after exposure and increased with time. Cells in the CA1 region seemed more vulnerable to the hormone than cells in other regions. At 1 week of exposure, the toxic effects were dose-dependent over the range of 0.1 pM to 0.1 microM, the minimum effective dose being 10 pM. The adverse effects were not induced either by the inactive fragment, PTH(39 - 84), or by an active fragment of PTH-related peptide (PTHrP(1 34)), an intrinsic ligand of the brain PTH receptor. The PTH(1 - 34)-induced adverse effects were significantly inhibited by co-administration of 10 microM nifedipine, an L-type Ca(2+) channel blocker, but not by co-administration of blockers of the other types of Ca(2+) channel. The present study demonstrates that sustained high levels of PTH in the brain might cause degeneration of specific brain regions due to Ca(2+) overloading via activation of dihydropyridine-sensitive Ca(2+) channels, and suggests that PTH may be a risk factor for senile dementia. British Journal of Pharmacology (2000) 129, 21 - 28 PMID- 10694199 TI - Stimulation of the ventral tegmental area enhances the effect of vasopressin on blood pressure in conscious rats. AB - The mesolimbic dopamine system projects to a large number of forebrain areas and plays an important role in the regulation of locomotor activity, cognition and reward. We previously found evidence for a functional interaction between the mesolimbic dopamine system and circulating vasopressin and the present study was performed to test the hypothesis that mesolimbic dopamine stimulation modulates the cardiovascular effects of vasopressin. Sprague-Dawley rats were stereotaxically implanted with a guide cannula into the region of origin of the mesolimbic system, the ventral tegmental area, and instrumented with catheters into the abdominal aorta and jugular vein. One week later, separate groups of conscious rats were injected intravenously with 1, 3 or 10 ng kg(-1) of arginine vasopressin or other vasopressor drugs before and after intra-ventral tegmental area injection of 10 nmol of neurotensin. Intra-ventral tegmental area injections of neurotensin had no significant effect on mean arterial pressure and heart rate but significantly potentiated the pressor response to intravenous administration of vasopressin when compared to saline-injections. However, the vasopressin induced bradycardia was unaffected. Intravenous pretreatment with raclopride blocked the ability of neurotensin, injected into the ventral tegmental area, to potentiate the vasopressin-induced pressor response. Intra ventral tegmental area injections of neurotensin had no effect on the pressor response and bradycardia induced by intravenous angiotensin II or methoxamine. In conclusion, these results suggest that the mesolimbic dopamine system, in addition to its well known role in the regulation of behaviour, modulates cardiovascular control by potentiating the effects of vasopressin on mean arterial pressure. British Journal of Pharmacology (2000) 129, 29 - 36 PMID- 10694201 TI - Effects of omega-conotoxin GVIA and diltiazem on double peaked vasoconstrictor responses to periarterial electric nerve stimulation in isolated canine splenic artery. AB - The actions of omega-conotoxin (omega-CTX) and diltiazem on adrenergic and purinergic components of double peaked vasoconstrictor responses to periarterial nerve stimulation have been investigated in the isolated, perfused canine splenic arterial preparation. Double peaked vasoconstrictions (biphases of vasoconstrictors) were consistently observed in the conditions of 30 s trains of pulses at 1 - 10 Hz frequencies. omega-CTX (1 - 30 nM) produced similar inhibitory effects on the first phase and second phase responses in a dose related manner. Thirty nM omega-CTX almost completely inhibited the biphasic vasoconstrictions at any used frequencies but did not affect the vasoconstrictor responses to exogenous applied ATP (0.01 - 1 micromol) and noradrenaline (0.03 - 3 nmol). Intraluminal application of a large dose of diltiazem (3 - 10 microM) also produced a dose-dependent inhibitory effect on biphasic vasoconstrictions at any used frequencies. Three microM diltiazem exerted rather a larger inhibitory effect on the second phase than the first phase response at low frequencies (1 - 3 Hz), but a similar inhibition on first and second phasic responses at high frequencies (6 - 10 Hz). An extremely high dose of diltiazem (10 microM) almost completely inhibited the biphasic vasoconstrictor responses to nerve stimulation, and slightly inhibited the contractile responses to exogenous applied ATP (0.01 - 1 micromol) and noradrenaline (0.03 - 3 nmol). The present results indicate that omega-CTX selectively acts prejunctionally to inhibit the release of transmitters from sympathetic nerve terminals, and omega-CTX-sensitive calcium channels may produce a parallel controlling of purinergic and adrenergic components of sympathetic cotransmission. A large dose of diltiazem has inhibitory effects on both prejunctional and postjunctional sympathetic co-transmission. British Journal of Pharmacology (2000) 129, 47 - 52 PMID- 10694200 TI - [(125)I]-GR231118: a high affinity radioligand to investigate neuropeptide Y Y(1) and Y(4) receptors. AB - GR231118 (also known as 1229U91 and GW1229), a purported Y(1) antagonist and Y(4) agonist was radiolabelled using the chloramine T method. [(125)I]-GR231118 binding reached equilibrium within 10 min at room temperature and remained stable for at least 4 h. Saturation binding experiments showed that [(125)I]-GR231118 binds with very high affinity (K(d) of 0.09 - 0.24 nM) in transfected HEK293 cells with the rat Y(1) and Y(4) receptor cDNA and in rat brain membrane homogenates. No specific binding sites could be detected in HEK293 cells transfected with the rat Y(2) or Y(5) receptor cDNA demonstrating the absence of significant affinity of GR231118 for these two receptor classes. Competition binding experiments revealed that specific [(125)I]-GR231118 binding in rat brain homogenates is most similar to that observed in HEK293 cells transfected with the rat Y(1), but not rat Y(4), receptor cDNA. Autoradiographic studies demonstrated that [(125)I]-GR231118 binding sites were fully inhibited by the Y(1) antagonist BIBO3304 in most areas of the rat brain. Interestingly, high percentage of [(125)I]-GR231118/BIBO3304-insensitive binding sites were detected in few areas. These [(125)I]-GR231118/BIBO3304-insensitive binding sites likely represent labelling to the Y(4) receptor subtype. In summary, [(125)I]-GR231118 is a new radiolabelled probe to investigate the Y(1) and Y(4) receptors; its major advantage being its high affinity. Using highly selective Y(1) antagonists such as BIBO3304 or BIBP3226 it is possible to block the binding of [(125)I]-GR231118 to the Y(1) receptor allowing for the characterization and visualization of the purported Y(4) subtype. British Journal of Pharmacology (2000) 129, 37 - 46 PMID- 10694202 TI - Effects of superoxide anion generators and thiol modulators on nitrergic transmission and relaxation to exogenous nitric oxide in the sheep urethra. AB - The effects of superoxide anion generators, the nitric oxide (NO) scavenger 2-(4 carboxyphenyl)-4,4,5,5-tetramethylimidazoine-1-oxyl 3-oxide (carboxy-PTIO), the specific guanylate cyclase inhibitor 1H-[1,2,4]-oxadiazole-[4,3-a]-quinoxalin-1 one (ODQ), and thiol modulating agents were investigated on relaxations induced by nitrergic stimulation and exogenous NO addition in the sheep urethra. Methylene blue (MB, 10 microM), pyrogallol (0.1 mM) and xanthine (X, 0.1 mM)/xanthine oxidase (XO, 0.1 u ml(-1)) inhibited NO-mediated relaxations, without affecting those induced by nitrergic stimulation. This resistance was not diminished following inhibition of endogenous Cu/Zn superoxide dismutase (Cu/Zn SOD) with diethyldithiocarbamic acid (DETCA, 3 mM), which almost abolished tissue SOD activity. Carboxy-PTIO (0.1 - 0.5 mM) inhibited NO-mediated relaxations but had no effect on responses to nitrergic stimulation, which were not changed by treatment with ascorbate oxidase (2 u ml(-1)). Relaxations to NO were reduced, but not abolished, by ODQ (10 microM), while nitrergic responses were completely blocked. The thiol modulators, ethacrynic acid (0.1 mM), diamide (1.5 mM), or 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB, 0. 5 mM), and subsequent treatment with dithiothreitol (DTT, 2 mM) had no effect on responses to nitrergic stimulation or NO. In contrast, N-ethylmaleimide (NEM, 0.2 mM) markedly inhibited both relaxations. L-cysteine (L-cys, 0.1 mM) had no effect on responses to NO, while it inhibited those to nitrergic stimulation, in a Cu/Zn SOD-independent manner. Our results do not support the view that the urethral nitrergic transmitter is free NO, and the possibility that another compound is acting as mediator still remains open. British Journal of Pharmacology (2000) 129, 53 - 62 PMID- 10694204 TI - Vascular endothelial growth factor attenuates trauma-induced injury in rats. AB - Endothelial dysfunction and loss of nitric oxide (NO) is an integral part of the initiation and maintenance of the inflammatory process such as that occurring in traumatic shock, and is considered responsible for much of the trauma induced microvascular injury. We investigated the effects of a vascular endothelial growth factor (VEGF) in a rat model of traumatic shock. Pentobarbital anaesthetized rats subjected to Noble-Collip drum trauma developed a shock state characterized by marked hypotension and a 93% mortality rate with a mean survival time of 108+/-10 min in 14 rats. Accompanying these effects was a significant degree of endothelial dysfunction and a markedly elevated intestinal myeloperoxidase (MPO) activity. Treatment with 125 microg kg(-1) VEGF administered intravenously 18 h pre-trauma, increased survival rate to 67% (P<0.01), and prolonged survival time to 252+/-24 min in 12 rats (P<0.01). VEGF also significantly preserved the endothelium-dependent relaxation to ACh indicating a preservation of endothelium-derived NO. Our results indicate that endothelial dysfunction with its accompanying loss of NO plays an important role in tissue injury associated with trauma, and that preservation of NO is beneficial in traumatic shock. The mechanisms of the protective effect of VEGF in trauma involves preservation of eNOS function and diminished neutrophil accumulation resulting in reduced neutrophil-mediated tissue injury. British Journal of Pharmacology (2000) 129, 71 - 76 PMID- 10694203 TI - Modulation of airway smooth muscle tone by protease activated receptor-1,-2,-3 and -4 in trachea isolated from influenza A virus-infected mice. AB - Relaxant and contractile effects of the tethered ligand domain sequences of murine PAR-1, PAR-2, PAR-3 and PAR-4, and of the proteases thrombin and trypsin were examined in mouse isolated tracheal preparations. The epithelium- and cyclo oxygenase-dependence of these effects and the potential modulatory effects of respiratory tract viral infection were also investigated. In carbachol-contracted preparations, trypsin, thrombin, and the tethered ligand domain sequences of murine PAR-1 (SFFLRN-NH(2)), PAR-2 (SLIGRL-NH(2)) and PAR-4 (GYPGKF-NH(2)), but not PAR-3 (SFNGGP-NH(2)), induced transient, relaxant responses that were abolished by the cyclo-oxygenase inhibitor indomethacin. Repeated administration of SFFLRN-NH(2), SLIGRL-NH(2) or GYPGKF-NH(2) (30 microM) was associated with markedly diminished relaxation responses (homologous desensitization), although there was no evidence of cross-desensitization between these peptides. The tethered ligand domain sequences for PAR-1 and PAR-4 induced a rapid, transient contractile response that preceded the relaxant response. Contractions were not inhibited by indomethacin and were not induced by either thrombin or trypsin. Influenza A virus infection did not significantly affect the responses induced by either the proteases or peptides. Furthermore, epithelial disruption caused by mechanical rubbing had no significant effect on responses to these PAR activators in preparations from either virus- or sham-infected mice. In summary, the proteases trypsin and thrombin, and peptide activators of PAR-1, PAR-2 and PAR-4 induced relaxant responses of mouse isolated tracheal smooth muscle preparations, which were mediated by a prostanoid, probably PGE(2). Interestingly, PAR-mediated relaxations were not significantly diminished following acute damage to the epithelium caused by mechanical rubbing and/or the respiratory tract viral pathogen, influenza A. British Journal of Pharmacology (2000) 129, 63 - 70. PMID- 10694205 TI - Bradyzide, a potent non-peptide B(2) bradykinin receptor antagonist with long lasting oral activity in animal models of inflammatory hyperalgesia. AB - Bradyzide is from a novel class of rodent-selective non-peptide B(2) bradykinin antagonists (1-(2-Nitrophenyl)thiosemicarbazides). Bradyzide has high affinity for the rodent B(2) receptor, displacing [(3)H]-bradykinin binding in NG108-15 cells and in Cos-7 cells expressing the rat receptor with K(I) values of 0.51+/ 0.18 nM (n=3) and 0.89+/-0.27 nM (n=3), respectively. Bradyzide is a competitive antagonist, inhibiting B(2) receptor-induced (45)Ca efflux from NG108-15 cells with a pK(B) of 8.0+/-0.16 (n=5) and a Schild slope of 1.05. In the rat spinal cord and tail preparation, bradyzide inhibits bradykinin-induced ventral root depolarizations (IC(50) value; 1.6+/-0.05 nM (n=3)). Bradyzide is much less potent at the human than at the rodent B(2) receptor, displacing [(3)H] bradykinin binding in human fibroblasts and in Cos-7 cells expressing the human B(2) receptor with K(I) values of 393+/-90 nM (n=3) and 772+/-144 nM (n=3), respectively. Bradyzide inhibits bradykinin-induced [(3)H]-inositol trisphosphate (IP(3)) formation with IC(50) values of 11.6+/-1.4 nM (n=3) at the rat and 2.4+/ 0.3 microM (n=3) at the human receptor. Bradyzide does not interact with a range of other receptors, including human and rat B(1) bradykinin receptors. Bradyzide is orally available and blocks bradykinin-induced hypotension and plasma extravasation. Bradyzide shows long-lasting oral activity in rodent models of inflammatory hyperalgesia, reversing Freund's complete adjuvant (FCA)-induced mechanical hyperalgesia in the rat knee joint (ED(50), 0.84 micromol kg(-1); duration of action >4 h). It is equipotent with morphine and diclofenac, and 1000 times more potent than paracetamol, its maximal effect exceeding that of the non steroidal anti-inflammatory drugs (NSAIDs). Bradyzide does not exhibit tolerance when administered over 6 days. In summary, bradyzide is a potent, orally active, antagonist of the B(2) bradykinin receptor, with selectivity for the rodent over the human receptor. British Journal of Pharmacology (2000) 129, 77 - 86 PMID- 10694206 TI - Sustained ethanol inhibition of native AMPA receptors on medial septum/diagonal band (MS/DB) neurons. AB - The direct impact of ethanol on native, non-NMDA glutamate receptors was examined in acutely isolated MS/DB neurons from rat. The impact of ethanol functional tolerance and physical dependence on non-NMDA receptor function was also determined. Non-NMDA receptors were defined pharmacologically as predominantly the AMPA subtype, because both AMPA- or kainate-activated currents were blocked by GYKI 52466, a selective AMPA receptor antagonist. The relative magnitude of potentiation of AMPA-activated currents by 10 or 100 microM cyclothiazide was consistent with recombinant AMPA flop-subtype receptors. Finally, the selective kainate receptor agonist, SYM 8021, induced little current in MS/DB neurons. AMPA receptor currents when activated by kainate were sensitive to ethanol, showing inhibition of approximately 5 - 50% when 10 - 300 mM ethanol and kainate were briefly co-applied (3 s). Ethanol (100 mM) also inhibited both the initial transient peak and sustained currents activated by AMPA. Inhibition was sustained during continuous ethanol superfusions of 5 min, suggesting a lack of acute tolerance to ethanol-induced AMPA receptor blockade. Rapid application of 3 - 3000 microM kainate activated concentration-dependent currents in MS/DB neurons from Control and Ethanol Dependent animals that were not significantly different. Also, direct ethanol inhibition (300 mM) of kainate-activated currents was not reduced by ethanol dependence, suggesting a lack of functional tolerance. These results suggest that native AMPA receptors on MS/DB neurons are inhibited by pharmacologically-relevant concentrations of ethanol. However, these receptors, unlike NMDA receptors, do not undergo adaptation with sustained ethanol exposure sufficient to induce physical dependence. British Journal of Pharmacology (2000) 129, 87 - 94 PMID- 10694207 TI - The metabolism of clomethiazole in gerbils and the neuroprotective and sedative activity of the metabolites. AB - A single dose of clomethiazole (600 micromol kg(-1) i.p.) has previously been shown to be neuroprotective in the gerbil model of global ischaemia. In gerbils, clomethiazole (600 micromol kg(-1)) injection produced a rapid appearance (peak within 5 min) of drug in plasma and brain and similar clearance (plasma t(1/2): 40 min) from both tissues. The peak brain concentration (226+/-56 nmol g(-1)) was 40% higher than plasma. One major metabolite, 5-(1-hydroxyethyl-2-chloro)-4 methylthiazole (NLA-715) and two minor metabolites 5-(1-hydroxyethyl)-4 methylthiazole (NLA-272) and 5-acetyl-4-methylthiazole (NLA-511) were detected in plasma and brain. Evidence suggested that clomethiazole is metabolized directly to both NLA-715 and NLA-272. Injection of NLA-715, NLA-272 or NLA-511 (each at 600 micromol kg(-1)) produced brain concentrations respectively 2.2, 38 and 92 times greater than seen after clomethiazole (600 micromol kg(-1)). Clomethiazole (600 micromol kg(-1)) injected 60 min after a 5 min bilateral carotid artery occlusion in gerbils attenuated the ischaemia-induced degeneration of the hippocampus by approximately 70%. The metabolites were not neuroprotective at this dose. In mice, clomethiazole (600 micromol kg(-1)) produced peak plasma and brain concentrations approximately 100% higher than in gerbils, drug concentrations in several brain regions were similar but 35% higher than plasma. Clomethiazole (ED(50): 180 micromol kg(-1)) and NLA-715 (ED(50): 240 micromol kg( 1)) inhibited spontaneous locomotor activity. The other metabolites were not sedative (ED(50) >600 micromol kg(-1)). These data suggest that the neuroprotective action of clomethiazole results from an action of the parent compound and that NLA-715 contributes to the sedative activity of the drug. British Journal of Pharmacology (2000) 129, 95 - 100 PMID- 10694208 TI - Lipophilization of somatostatin analog RC-160 with long chain fatty acid improves its antiproliferative and antiangiogenic activity in vitro. AB - The therapeutic potential of the somatostatin analogue RC-160 having antiproliferative activity, is limited by its short serum half life. To overcome this limitation, fatty acids namely butanoic acid and myristic acid were conjugated to the N-terminal residue of RC-160. The lipophilized derivatives of RC-160 were synthesized, purified by reverse phase HPLC and characterized by ES mass spectroscopy. The antiproliferative activity of lipophilized derivatives of RC-160 on the growth of MIA-PaCa2 (human pancreatic carcinoma), DU145 (human prostate carcinoma), ECV304 (human umbilical chord endothelioma), as well as their antiangiogenic activity was evaluated in vitro. The relative stability of myristoyl-RC-160 towards degradation by proteases and serum was also determined. Myristoyl-RC-160 exhibited significantly higher antiproliferative efficacy than RC-160, on the above cell lines (P<0.01). Receptor binding assays, demonstrated that the affinity of RC-160 towards somatostatin receptors remains unaltered by myristoylation. Unlike RC-160, the myristoylated derivative was found to have significantly greater resistance to protease and serum degradation (P<0.01). Myristoyl-RC-160 exhibited significantly greater antiproliferative activity on ECV304, than RC-160 (P<0.01). Myristoyl RC-160 could also inhibit capillary tube formation more efficiently than RC-160 in a dose dependent manner, suggesting that it possessed enhanced antiangiogenic activity in vitro (P<0.001). Lipophilization of RC-160 with long chain fatty acids like myristic acid endows it with improved antiproliferative and antiangiogenic activity, stability and therapeutic index. British Journal of Pharmacology (2000) 109, 101 - 109 PMID- 10694209 TI - The peripheral antinociceptive effect induced by morphine is associated with ATP sensitive K(+) channels. AB - The effect of several K(+) channel blockers such as glibenclamide, tolbutamide, charybdotoxin (ChTX), apamin, tetraethylammonium (TEA), 4-aminopyridine (4-AP) and cesium on the peripheral antinociceptive effect of morphine was evaluated by the paw pressure test in Wistar rats. The intraplantar administration of a carrageenan suspension (250 microg) resulted in an acute inflammatory response and a decreased threshold to noxious pressure. Morphine administered locally into the paw (25, 50, 100 and 200 microg) elicited a dose-dependent antinociceptive effect which was demonstrated to be mediated by a peripheral site up to the 100 microg dose. The selective blockers of ATP-sensitive K(+) channels glibenclamide (20, 40 and 80 microg paw(-1)) and tolbutamide (40, 80 and 160 microg paw(-1)) antagonized the peripheral antinociception induced by morphine (100 microg paw( 1)). This effect was unaffected by ChTX (0. 5, 1.0 and 2.0 microg paw(-1)), a large conductance Ca(2+)-activated K(+) channel blocker, or by apamin (2.5, 5.0 and 10.0 microg paw(-1)), a selective blocker of a small conductance Ca(2+) activated K(+) channel. Intraplantar administration of the non-specific K(+) channel blockers TEA (160, 320 and 640 microg), 4-AP (10, 50 and 100 microg) and cesium (125, 250 and 500 microg) also did not modify the peripheral antinociceptive effect of morphine. These results suggest that the peripheral antinociceptive effect of morphine may result from activation of ATP-sensitive K(+) channels, which may cause a hyperpolarization of peripheral terminals of primary afferents, leading to a decrease in action potential generation. In contrast, large conductance Ca(2+)-activated K(+) channels, small conductance Ca(2+)-activated K(+) channels as well as voltage-dependent K(+) channels appear not to be involved in this transduction pathway. British Journal of Pharmacology (2000) 129, 110 - 114 PMID- 10694211 TI - Effects of diadenosine polyphosphates (Ap(n)As) and adenosine polyphospho guanosines (Ap(n)Gs) on rat mesenteric artery P2X receptor ion channels. AB - Diadenosine polyphosphates (Ap(n)As, n=3 - 7) and adenosine polyphospho guanosines (Ap(n)Gs, n=3 - 6) are naturally occurring vasoconstrictor substances found in platelets. These vasoconstrictor actions are thought to be mediated through the activation of P2X receptors for ATP. The effects of Ap(n)As and Ap(n)Gs at P2X receptors on rat mesenteric arteries were determined in contraction studies and using the patch clamp technique on acutely dissociated artery smooth muscle cells. P2X(1) receptor immunoreactivity was detected in the smooth muscle layer of artery rings. The sensitivity to alpha,beta-methylene ATP and desensitizing nature of rat mesenteric artery P2X receptors correspond closely to those of recombinant P2X(1) receptors. Ap(4)A, Ap(5)A and Ap(6)A evoked concentration dependent P2X receptor inward currents which desensitized during the application of higher concentrations of agonist. The agonist order of potency was Ap(5)A> or = Ap(6)A> or = Ap(4)A >> Ap(3)A. Ap(2)A and Ap(7)A were ineffective. Similar results were obtained in contraction studies except for Ap(7)A which evoked a substantial contraction. Ap(n)Gs (n=2 - 6)(30 microM) evoked P2X receptor inward currents in mesenteric artery smooth muscle cells. Ap(n)Gs (n=4 - 6) were less effective than the corresponding Ap(n)A. This study shows that at physiologically relevant concentrations Ap(n)As and Ap(n)Gs can mediate contraction of rat mesenteric arteries through the activation of P2X(1) like receptors. However the activity of the longer chain polyphosphates (n=6 - 7) may be overestimated in whole tissue studies due to metabolic breakdown to yield the P2X receptor agonists ATP and adenosine tetraphosphate. British Journal of Pharmacology (2000) 129, 124 - 130 PMID- 10694210 TI - Functional neuroimaging of cognition impaired by a classical antihistamine, d chlorpheniramine. AB - Antihistamine induced cognitive decline was evaluated using positron emission tomography (PET) measurement of histamine H1 receptor (H1R) occupancy and regional cerebral blood flow (rCBF). Cognitive performance in attention-demanding task deteriorated dose-dependently and the effects were statistically significant after the treatment of 2 mg of d-chlorpheniramine. There was no significant change in subjective sleepiness in the same dose. The regional blockade of H1R was observed mainly in the frontal, temporal and anterior cingulate cortices, and the intravenous administration of d-chlorpheniramine as a therapeutic dose (2 mg) blocked over 60% of H1R in the frontal cortices. The results from activation study using visual discrimination tasks demonstrated that enhanced activity in the right prefrontal and anterior cingulate cortices as well as a decreased activity in the left temporal and frontal cortices and midbrain after the treatment of d-chlorpheniramine. There were no changes in global CBF for the subjects treated with 2 mg d-chlorpheniramine (pre; 44.8+/-3.3 ml dl(-1) min(-1) vs post; 44.4+/-4.7 ml dl(-1) min(-1)). The results indicated that the attention system of human brain could be altered by therapeutic doses of H1R antagonists. These findings provide the information as to the potential risk of antihistamines in our daily activities. British Journal of Pharmacology (2000) 129, 115 - 123 PMID- 10694212 TI - Pharmacologic characterization of the oxytocin receptor in human uterine smooth muscle cells. AB - [(3)H]-oxytocin was used to characterize the oxytocin receptor found in human uterine smooth muscle cells (USMC). Specific binding of [(3)H]-oxytocin to USMC plasma membranes was dependent upon time, temperature and membrane protein concentration. Scatchard plot analysis of equilibrium binding data revealed the existence of a single class of high-affinity binding sites with an apparent equilibrium dissociation constant (K(d)) of 0.76 nM and a maximum receptor density (B(max)) of 153 fmol mg(-1) protein. The Hill coefficient (n(H)) did not differ significantly from unity, suggesting binding to homogenous, non interacting receptor populations. Competitive inhibition of [(3)H]-oxytocin binding showed that oxytocin and vasopressin (AVP) receptor agonists and antagonists displaced [(3)H]-oxytocin in a concentration-dependent manner. The order of potencies for peptide agonists and antagonists was: oxytocin>[Asu(1,6)] oxytocin>AVP= atosiban>d(CH(2))(5)Tyr(Me)AVP>[Thr(4),Gly(7)]-oxytocin>dDAVP, and for nonpeptide antagonists was: L-371257>YM087>SR 49059>OPC-21268>SR 121463A>OPC 31260. Oxytocin significantly induced concentration-dependent increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) and hyperplasia in USMC. The oxytocin receptor antagonists, atosiban and L-371257, potently and concentration dependently inhibited oxytocin-induced [Ca(2+)](i) increase and hyperplasia. In contrast, the V(1A) receptor selective antagonist, SR 49059, and the V(2) receptor selective antagonist, SR 121463A, did not potently inhibit oxytocin induced [Ca(2+)](i) increase and hyperplasia. The potency order of antagonists in inhibiting oxytocin-induced [Ca(2+)](i) increase and hyperplasia was similar to that observed in radioligand binding assays. In conclusion, these data provide evidence that the high-affinity [(3)H]-oxytocin binding site found in human USMC is a functional oxytocin receptor coupled to [Ca(2+)](i) increase and cell growth. Thus human USMC may prove to be a valuable tool in further investigation of the physiologic and pathophysiologic roles of oxytocin in the uterus. British Journal of Pharmacology (2000) 129, 131 - 139 PMID- 10694213 TI - Involvement of cyclic AMP - PKA pathway in VIP-induced, charybdotoxin-sensitive relaxation of longitudinal muscle of the distal colon of Wistar-ST rats. AB - The intracellular mechanism of vasoactive intestinal peptide (VIP)-induced, charybdotoxin (ChTx)-sensitive relaxation of longitudinal muscle of the distal colon of Wistar-ST rats was studied. A single pulse or 100 pulses at 10 Hz of electrical field stimulation (EFS) induced rapid transient relaxation or that with a subsequent contraction of the longitudinal muscle in the presence of atropine and guanethidine, respectively. Rp-8 bromo cAMPS, an inhibitor of cyclic AMP dependent protein kinase (PKA), at 30 microM inhibited the relaxations induced by EFS with a single or 100 pulses maximally by about 80 or 60%, respectively. It also inhibited VIP (300 nM)-induced relaxation by 82%. VIP (100 nM - 1 microM) increased the cyclic AMP content of longitudinal muscle myenteric plexus preparations obtained from the distal colon. ChTx at 100 nM almost completely inhibited 8 bromo cyclic AMP-induced relaxation of the distal segments. EFS with two or three pulses at 10 Hz induced inhibitory junction potentials consisting of two phases, rapid and subsequent slow hyperpolarization in the membrane potential of longitudinal smooth muscle cells. Rp-cAMPS, another inhibitor of PKA, inhibited the delayed slow hyperpolarization. It also inhibited the exogenously added VIP-induced hyperpolarization of the cell membrane. Thus, the present study suggests that activation of PKA via activation of VIP receptors is associated with activation of ChTx-sensitive K(+) channels in relaxation of longitudinal muscle of the distal colon of Wistar-ST rats. British Journal of Pharmacology (2000) 129, 140 - 146 PMID- 10694214 TI - Effects of nicotine and chlorisondamine on cerebral glucose utilization in immobilized and freely-moving rats. AB - Chlorisondamine blocks central nicotinic receptors for many weeks via an unknown mechanism. Intracerebroventricular administration of [(3)H]-chlorisondamine in rats results in an anatomically restricted and persistent intracellular accumulation of radioactivity. The initial aim of the present study was to test whether nicotinic receptor antagonism by chlorisondamine is also anatomically restricted. Male adult rats were pretreated several times with nicotine to avoid the disruptive effects of the drug seen in drug-naive animals. They then received chlorisondamine (10 microg i. c.v.) or saline, and local cerebral glucose utilization (LCGU) was measured 4 weeks later after acute nicotine (0.4 mg kg(-1) s.c.) or saline administration. During testing, rats were partially immobilized. Nicotine significantly increased LCGU in the anteroventral thalamus and in superior colliculus. Chlorisondamine completely blocked the first of these effects. Chlorisondamine significantly reduced LCGU in the lateral habenula, substantia nigra pars compacta, ventral tegmental area, and cerebellar granular layer. The second experiment was of similar design, but the rats were not pre exposed to nicotine, and were tested whilst freely-moving. Acute nicotine significantly increased LCGU in anteroventral thalamus, superior colliculus, medial habenula and dorsal lateral geniculate. Overall, however, nicotine significantly decreased LCGU. Most or all of the central effects of nicotine on LCGU were reversed by chlorisondamine given 4 weeks beforehand. These findings suggest that chlorisondamine blocks nicotinic effects widely within the brain. They also indicate that in freely-moving rats, nicotine can reduce or stimulate cerebral glucose utilization, depending on the brain area. British Journal of Pharmacology (2000) 129, 147 - 155 PMID- 10694215 TI - Biphasic effects of NMDA on the motility of the rat portal vein. AB - The effect of NMDA on the motility of the rat portal vein was studied in an isolated preparation. NMDA induced a concentration-dependent (10(-7) - 10(-4) M) increase of the contraction frequency (maximum increase, 148+/-6% of control at NMDA 10(-4) M). The NMDA-induced excitatory response was prevented by the competitive NMDA receptor antagonists (+/-)-2-Amino-5-phosphonopentanoic acid (AP 5, 5x10(-4) M) or (RS)-3-(2-carboxypiperazine-4-yl) propyl-1-phosphonic acid (CPP, 10(-4) M). Tetrodotoxin (TTX, 10(-6) M) or atropine (10(-4) M) abolished the NMDA-induced increase of the portal vein motility and reversed the excitatory effect to a concentration-dependent inhibition (maximum inhibition, 52+/-8 and 29+/-7% of controls, respectively, at NMDA 10(-3) M). Removal of the endothelium abolished the NMDA-induced inhibitory response. Sodium nitroprusside concentration-dependently (10(-7) - 10(-5) M) inhibited the portal vein motility, while L-N(G)-nitro-arginine methyl ester (L-NAME, 10(-4) M) reversed the inhibitory effect of NMDA (in the presence of TTX), restoring the portal vein spontaneous activity to control values. These results show that NMDA modulates the portal vein motility in a biphasic manner: via indirect activation, through prejunctional NMDA receptors presumably located on intrinsic excitatory neuronal afferences, or via direct inhibition, through endothelial NMDA receptors activating the nitric oxide pathway. Overall these findings support the hypothesis of the existence of a peripheral glutamatergic innervation modulating the contractile activity of the rat portal vein. British Journal of Pharmacology (2000) 129, 156 - 162 PMID- 10694216 TI - Effect of 18beta-glycyrrhetinic acid on electromechanical coupling in the guinea pig renal pelvis and ureter. AB - We have tested the effect of the gap junction inhibitor, 18beta-glycyrrhetinic acid (18betaGA) on electromechanical coupling in the guinea-pig renal pelvis and ureter by the sucrose gap technique. In the ureter 18betaGA (3 - 30 microM) produced a concentration-dependent inhibition of the spike component of the action potential (AP) and reduced contraction evoked by electrical stimulation. Neurokinin A (NKA) produced a slow depolarization with superimposed APs and phasic contractions of the ureter. 18betaGA (30 microM) markedly inhibited the depolarization and APs evoked by NKA. However the contractile response was more sustained in the presence than in the absence of 18betaGA. At 100 microM, 18betaGA inhibited the mechanical responses to NKA. KCl (80 mM) produced APs and phasic contractions followed by sustained depolarization and tonic contraction. At 30 microM 18betaGA markedly inhibited the KCl-evoked APs and phasic contractions without affecting the sustained responses. At 100 microM 18betaGA inhibited the tonic contraction to KCl. In the renal pelvis 18betaGA (30 microM) inhibited the amplitude of pacemaker potentials and accompanying contractions and induced the appearance of low-amplitude APs not associated with contraction. We conclude that, up to 30 microM, the action of 18betaGA is consistent with an inhibition of cell-to-cell electrical coupling via gap junctions. The single-unit character of smooth muscles in the guinea-pig upper urinary tract is partly converted to a multi-unit pattern. At high concentrations 18betaGA possesses non specific effects which limit its usefulness as a tool for studying the role of gap junctions in smooth muscles. British Journal of Pharmacology (2000) 129, 163 169 PMID- 10694217 TI - Contraction to big endothelin-1, big endothelin-2 and big endothelin-3, and endothelin-converting enzyme inhibition in human isolated bronchi. AB - All three endothelin precursor peptides, i.e. big endothelin-1 (big ET-1), big endothelin-2 (big ET-2) and big endothelin-3 (big ET-3), produced contractile responses in human isolated bronchi, demonstrating the presence of functional endothelin-converting enzyme (ECE) in this tissue. The maximal contractile responses were equal to 108.4+/-8.0% (0.1 microM big ET-1; n=4), 85.2+/-11.8% (0.1 microM big ET-2; n=7) and 43.0+/-7.2% (0.1 microM big ET-3; n=5) of the reference response to acetylcholine (1 mM). The response to big ET-1 (0.1 microM), but not endothelin-1 (ET-1, 0.1 microM), was diminished after overnight storage of the tissue at 4 degrees C, demonstrating instability of the enzyme. The responses to all three big-endothelins were significantly inhibited, by the ECE inhibitors CGS 26393 and CGS 26303, in a concentration-related manner. The responses to the mature peptides ET-1, endothelin-2 (ET-2), and endothelin-3 (ET 3) were unaffected by CGS 26393 and CGS 26303. Phosphoramidon (10 microM) also produced an inhibition of the response to big ET-1 that was equivalent to that produced by CGS 26393 (10 microM). Combination of CGS 26393 (10 microM) and phosphoramidon (10 microM) did not produce an additive inhibition. These results demonstrate the presence of functional ECE for all three big endothelins in human bronchus and inhibition of the enzyme by newly developed orally active ECE inhibitors, as well as phosphoramidon. British Journal of Pharmacology (2000) 129, 170 - 176 PMID- 10694218 TI - Evidence that activation of central 5-HT(2B) receptors causes renal sympathoexcitation in anaesthetized rats. AB - The effects of injections i.c.v. of alpha-methyl-5-(2-thienylmethoxy)-1H-indole-3 ethanamine (BW723C86; 0.02 - 2 micromol kg(-1)), a 5-HT(2B) receptor agonist, on renal sympathetic and phrenic nerve activity, mean arterial blood pressure and heart rate were investigated in alpha-chloralose anaesthetized rats pretreated with a peripherally acting 5-HT(2) receptor antagonist. BW723C86 i.c.v. caused a dose-related increase in renal nerve activity reaching a maximum of 67+/-6%, which at the highest dose was associated with a small and maintained fall in mean arterial blood pressure of 7+/-3 mmHg. These changes were not associated with any significant changes in heart rate or phrenic nerve activity. BW723C86-evoked increases in renal nerve activity and hypotension were attenuated by pretreatment (i.c.v.) with SB204741 (300 nmol kg(-1); a 5-HT(2B) receptor antagonist) but not by the same dose (i.c.v.) of ketanserin (a 5-HT(2A) receptor antagonist) or RS102221 (a 5-HT(2C) receptor antagonist). None of these antagonists alone had any effect on the variables being measured. It is concluded that central 5-HT(2B) receptors may play a selective role in the control of sympathetic supply to the kidney, which could be important in the central mechanisms involved in blood volume regulation. British Journal of Pharmacology (2000) 129, 177 - 183 PMID- 10694219 TI - Nitric oxide, prostanoid and non-NO, non-prostanoid involvement in acetylcholine relaxation of isolated human small arteries. AB - The main purpose of the study was to clarify to which extent nitric oxide (NO) contributes to acetylcholine (ACh) induced relaxation of human subcutaneous small arteries. Arterial segments were mounted in myographs for recording of isometric tension, NO concentration and smooth muscle membrane potential. In noradrenaline contracted arteries, ACh induced endothelium-dependent relaxations. The NO synthase inhibitor, N(G)-nitro-L-arginine (L-NOARG) had a small significant effect on the concentration-response curves for ACh, and in the presence of L NOARG, indomethacin only caused a small additional rightward shift in the ACh relaxation. The NO scavenger, oxyhaemoglobin attenuated relaxations for ACh and for the NO donor S-nitroso-N-acetylpenicillamine (SNAP). Inhibition of guanylyl cyclase with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), and inhibition of protein kinase G with beta-phenyl-1, N2-etheno-8-bromoguanosine- 3', 5'- cyclic monophosphorothioate, Rp-isomer, slightly attenuated ACh relaxation, but abolished SNAP induced relaxation. ACh induced relaxation without increases in the free NO concentration. In contrast, for equivalent relaxation, SNAP increased the NO concentration 32+/-8 nM. ACh hyperpolarized the arterial smooth muscle cells with 11.4+/-1.3 mV and 10.5+/-1.3 mV in the absence and presence of L NOARG, respectively. SNAP only elicited a hyperpolarization of 1.6+/-0.9 mV. In the presence of indomethacin and L-NOARG, ACh relaxation was almost unaffected by lipoxygenase inhibition with nordihydroguaiaretic acid, or cytochrome P450 inhibition with 17-octadecynoic acid or econazole. ACh relaxation was strongly reduced by the combination of charybdotoxin and apamin, but small increments in the extracellular potassium concentration induced no relaxations. The study demonstrates that the NO/L-arginine pathway is present in human subcutaneous small arteries and to a limited extent is involved in ACh induced relaxation. The study also suggests a small contribution of arachidonic acid metabolites. However, ACh relaxation is mainly dependent on a non-NO, non-prostanoid endothelium dependent hyperpolarization. British Journal of Pharmacology (2000) 129, 184 - 192 PMID- 10694220 TI - Role of tachykinin NK2 receptors in normal and altered rectal sensitivity in rats. AB - Irritable bowel syndrome is characterized by visceral hyperalgesia commonly associated with stress and inflammatory processes. We investigated the role of tachykinin NK2 receptors in the ability of trinitrobenzenesulphonic acid (TNBS) and stress to enhance the sensitivity of the rat rectum to distension using a selective tachykinin NK2 receptor antagonist (MEN 11420). Rats were fitted with electrodes implanted in the striated muscles of the abdomen. Rectal distension (RD) was performed with a balloon inflated by steps of 0.4 ml from 0 to 1.6 ml. Five groups were submitted to RD performed 3 days before and after intrarectal instillation of TNBS. Fifteen minutes before RD, rats were treated with saline or MEN 11420 (5 - 100 microg kg(-1) i.v.). Two other groups, submitted to 2 h restraint or sham stress sessions were randomly treated i.v. with saline or MEN 11420 (10 - 200 microg kg(-1)) prior to RD applied 20 min later. The basal response to RD was characterized by a significant increase in the number of abdominal contractions. This response occurred with a threshold volume of 0.8 ml and was dose-dependently reduced by MEN 11420 (5 - 100 microg kg(-1) i.v.). Rectal inflammation lowered the volume of distension producing abdominal contractions to 0.4 ml (allodynia). This effect was either reduced or suppressed by MEN 11420. A similar allodynia was observed after a stress session and this effect was reduced (49%) or suppressed by MEN 11420 at 200 and 100 microg kg(-1), respectively. Tachykinin NK2 receptors are involved in rectal hypersensitivity associated with inflammation and stress. British Journal of Pharmacology (2000) 129, 193 - 199 PMID- 10694221 TI - Differential effects of the tricyclic antidepressant amoxapine on glycine uptake mediated by the recombinant GLYT1 and GLYT2 glycine transporters. AB - We examined the effects of nine different tricyclic antidepressant drugs on the glycine uptake mediated by the glycine transporter 1b (GLYT1b) and glycine transporter 2a (GLYT2a) stably expressed in human embryonic kidney 293 cells. Desipramine, imipramine, clomipramine, nomifensine and mianserin had no effect on the activity of the glycine transporters. Doxepin, amitriptyline and nortriptyline inhibited the two transporter subtypes to a similar extent. Amoxapine displayed a selective inhibition of GLYT2a behaving as a 10 fold more efficient inhibitor of this isoform than of GLYT1b. Kinetic analysis of the initial rates of glycine uptake by GLYT2a as a function of either glycine, chloride or sodium concentration, in the absence and presence of amoxapine indicated that amoxapine behaved as a competitive inhibitor of both glycine and chloride and a mixed-type inhibitor with respect to sodium. A kinetic model was developed which explains adequately these data, and gives information about the order of binding of sodium and chloride ions to GLYT2a. Our results may contribute to the development of the glycine transporter pharmacology. Additionally, the inhibition of the glycine uptake by GLYT2 is suggested to have some role in the sedative and psychomotor side effects of amoxapine. British Journal of Pharmacology (2000) 129, 200 - 206 PMID- 10694222 TI - Nitric oxide synthase expression, enzyme activity and NO production during angiogenesis in the chick chorioallantoic membrane. AB - In order to elucidate further the role of nitric oxide (NO) as an endogenous antiangiogenic mediator, mRNA expression of inducible nitric oxide synthase (iNOS), enzyme activity and production of NO were determined in the chick chorioallantoic membrane (CAM), an in vivo model of angiogenesis. In this model, maximum angiogenesis is reached between days 9 - 12 of chick embryo development. After that period, vascular density remains constant. Inducible NO synthase (iNOS) mRNA expression, determined by reverse transcriptase polymerase chain reaction (RT - PCR), increased from the 8th day reaching a maximum (70% increase) at days 10 - 11. NO synthase activity, determined as citrulline formation in the presence of calcium, also increased from day 8 reaching a maximum around day 10 (100% increase). Similar results were obtained in the absence of calcium suggesting that the NOS determined was the inducible form. Nitric oxide production, determined as nitrites, increased from day 8 reaching a maximum around day 10 (64% increase) and remaining stable at day 13. Finally, the bacterial lipopolysaccharide LPS (which activates transcriptionally iNOS), inhibited dose dependently angiogenesis in the CAM. These results in connection with previous findings from this laboratory, showing that NO inhibits angiogenesis in the CAM, suggest that increases in iNOS expression, enzyme activity and NO production closely parallel the progression of angiogenesis in the CAM, thus providing an endogenous brake to control this process. British Journal of Pharmacology (2000) 129, 207 - 213 PMID- 10694225 TI - The endogenous lipid anandamide is a full agonist at the human vanilloid receptor (hVR1). AB - The endogenous cannabinoid anandamide was identified as an agonist for the recombinant human VR1 (hVR1) by screening a large array of bioactive substances using a FLIPR-based calcium assay. Further electrophysiological studies showed that anandamide (10 or 100 microM) and capsaicin (1 microM) produced similar inward currents in hVR1 transfected, but not in parental, HEK293 cells. These currents were abolished by capsazepine (1 microM). In the FLIPR anandamide and capsaicin were full agonists at hVR1, with pEC(50) values of 5. 94+/-0.06 (n=5) and 7.13+/-0.11 (n=8) respectively. The response to anandamide was inhibited by capsazepine (pK(B) of 7.40+/-0.02, n=6), but not by the cannabinoid receptor antagonists AM630 or AM281. Furthermore, pretreatment with capsaicin desensitized the anandamide-induced calcium response and vice versa. In conclusion, this study has demonstrated for the first time that anandamide acts as a full agonist at the human VR1. PMID- 10694226 TI - Curcumin prevents adriamycin nephrotoxicity in rats. AB - The present study investigated the effect of curcumin on adriamycin (ADR) nephrosis in rats. The results indicate that ADR-induced kidney injury was remarkably prevented by treatment with curcumin. Treatment with curcumin markedly protected against ADR-induced proteinuria, albuminuria, hypoalbuminaemia and hyperlipidaemia. Similarly, curcumin inhibited ADR-induced increase in urinary excretion of N-acetyl-beta-D-glucosaminidase (a marker of renal tubular injury), fibronectin and glycosaminoglycan and plasma cholesterol. Curcumin restored renal function in ADR rats, as judged by the increase in GFR. The data also demonstrated that curcumin protected against ADR-induced renal injury by suppressing oxidative stress and increasing kidney glutathione content and glutathione peroxidase activity. In like manner, curcumin abolished ADR stimulated kidney microsomal and mitochondrial lipid peroxidation. These data suggest that administration of curcumin is a promising approach in the treatment of nephrosis caused by ADR. PMID- 10694227 TI - A novel anionic conductance affects action potential duration in isolated rat ventricular myocytes. AB - Effects of extracellular anions were studied in electrophysiological experiments on freshly isolated rat ventricular myocytes. Under current-clamp, action potential duration (APD) was prolonged by reducing the extracellular Cl(-) concentration and shortened by replacement of extracellular Cl(-) with I(-). Under voltage-clamp, membrane potential steps or ramps evoked an anionic background current (I(AB)) carried by either Cl(-), Br(-), I(-) or NO(3)(-). Activation of I(AB) was Ca(2+)- and cyclic AMP-independent, and was unaffected by cell shrinkage. I(AB) was insensitive to stilbene and fenamate anion transport blockers at concentrations that inhibit Ca(2+)-, cyclic AMP- and swelling activated Cl(-) currents in ventricular cells of other mammals. These results suggest that I(AB) may be carried by a novel class of Cl(-) channel. Correlation of anion substitution experiments on membrane current and action potentials revealed that I(AB) could play a major role in controlling rat ventricular APD. These findings have important implications for those studying cardiac Cl(-) channels as potential targets for novel antiarrythmic agents. PMID- 10694228 TI - Enhanced nociception by exogenous and endogenous substance P given into the spinal cord in mice lacking NR(2)A/epsilon(1), an NMDA receptor subunit. AB - In capsaicin-pretreated mice, the nociceptive responses induced by intrathecally (i.t.) administered substance P (SP) were enhanced by N-methyl-D-aspartate (NMDA) type receptor antagonists, dizocilpine (MK801) and D-2-amino-5 phosphonopentanoate (D-AP5) in a dose-dependent manner. Similar enhancement of SP induced nociception was also observed in mice lacking the NMDA-type glutamate receptor NR2A/epsilon(1) subunit gene (GluRepsilon(1)(-/-) mice). On the other hand, GluRepsilon(1)(-/-) mice showed a marked enhancement of the peripheral nociceptive responses induced by intraplantar (i.pl.) injection of SP and bradykinin (BK). As the nociceptive responses to SP and BK (i.pl.) were both antagonized by CP-99994, an neurokinin(1) (NK(1)) antagonist (i.t.), these results suggest that GluRepsilon(1) receptor may play an inhibitory role in the downstream mechanisms of primary nociceptive SP neurones, possibly through activation of unidentified inhibitory neurones. PMID- 10694229 TI - Adenosine induces cyclic-AMP formation and inhibits endothelin-1 production/secretion in guinea-pig tracheal epithelial cells through A(2B) adenosine receptors. AB - 1. The adenosine receptor subtype mediating adenosine 3' : 5'-cyclic monophosphate (cyclic AMP) formation and the effect of its activation on endothelin-1 (ET-1) secretion were studied in primary cultures of tracheal epithelial cells. 2. Adenosine analogues showed the following rank order of potency (pD(2) value) and intrinsic activity on the generation of cyclic AMP by tracheal epithelial cells: 5'-N-ethylcarboxyamidoadenosine (NECA, A(1)/A(2A)/A(2B), pD(2): 5.44+/-0.16)>adenosine (ADO, non selective, pD(2): 4.99+/-0. 09; 71+/-9% of NECA response) >/=2-Cl-adenosine (2CADO, non selective, pD(2): 4.72+/-0.14; 65+/-9% of NECA response)>>>CGS21680 (A(2A); inactive at up to 100 microM). 3. Cyclic AMP formation stimulated by NECA in guinea-pig tracheal epithelial cells was inhibited by adenosine receptor antagonist with the following order of apparent affinity (pA(2) value): Xanthine amine congeners (XAC, A(2A)/A(2B), 7.89+/-0.22)>CGS15943 (A(2A)/A(2B), 7.24+/-0. 26)>ZM241385 (A(2A), 6.69+/-0.14)>DPCPX (A(1), 6.51+/-0. 14)>3n-propylxanthine (weak A(2B), 4.30+/-0.10). This rank order of potency is typical for A(2B)-adenosine receptor. 4. Adenosine decreased basal and LPS-stimulated irET production in a concentration-dependent manner. Moreover, NECA but not CGS21680 inhibited LPS induced irET production. 5. The inhibitory effect of NECA on LPS-induced irET production was reversed by XAC (pA(2)=8.84+/-0. 12) and DPCPX (pA(2)=8.10+/ 0.22). 6. These results suggested that adenosine increased cyclic AMP formation and inhibited irET production/secretion by guinea-pig tracheal epithelial cells through the activation of a functional adenosine receptor that is most likely the A(2B) subtype. This adenosine receptor may be involved in the regulation of the level of ET-1 production/secretion by guinea-pig tracheal epithelial cells in physiological as well as in pathophysiological conditions. PMID- 10694230 TI - P-glycoprotein- and mrp2-mediated octreotide transport in renal proximal tubule. AB - 1. Transepithelial transport of a fluorescent derivative of octreotide (NBD octreotide) was studied in freshly isolated, functionally intact renal proximal tubules from killifish (Fundulus heteroclitus). 2. Drug accumulation in the tubular lumen was visualized by means of confocal microscopy and was measured by image analysis. Secretion of NBD-octreotide into the tubular lumen was demonstrated and exhibited the all characteristics of specific and energy dependent transport. Steady state luminal fluorescence averaged about five times cellular fluorescence and was reduced to cellular levels when metabolism was inhibited by NaCN. 3. NBD-octreotide secretion was inhibited in a concentration dependent manner by unlabelled octreotide, verapamil and leukotriene C(4) (LTC(4)). Conversely, unlabelled octreotide reduced in a concentration dependent manner the p-glycoprotein (Pgp)-mediated secretion of a fluorescent cyclosporin A derivative (NBDL-CS) and the mrp2-mediated secretion of fluorescein methotrexate (FL-MTX). 4. This inhibition was not due to impaired metabolism or toxicity since octreotide had no influence on the active transport of fluorescein (FL), a substrate for the classical renal organic anion transport system. 5. The data are consistent with octreotide being transported across the brush border membrane of proximal kidney tubules by both Pgp and mrp2. PMID- 10694231 TI - Mechanisms of AVP-induced glucagon release in clonal alpha-cells in-R1-G9: involvement of Ca(2+)-dependent and -independent pathways. AB - 1. The mechanisms underlying AVP-induced increase in [Ca(2+)](i) and glucagon release in clonal alpha-cells In-R1-G9 were investigated. 2. AVP increased [Ca(2+)](i) and glucagon release in a concentration-dependent manner. After the administration of AVP, glucagon was released within 30 s, quickly reached the maximum within 2 min, and maintained a steady-state concentration for at least 15 min. 3. In Ca(2+)-containing medium, AVP increased [Ca(2+)](i) in a biphasic pattern; a peak followed by a sustained plateau. In Ca(2+)-free medium, the Ca(2+) response to AVP became monophasic with lower amplitude and no plateau. Both the basal and AVP-induced glucagon releases were lower in the absence than in the presence of extracellular Ca(2+). When [Ca(2+)](i) was stringently deprived by BAPTA, a Ca(2+) chelator, AVP still significantly increased glucagon release. 4. Pretreatment with thapsigargin, a microsomal Ca(2+) ATPase inhibitor, abolished both the Ca(2+) peak and sustained plateau. 5.AVP increased intracellular concentration of IP(3). 6. U-73122 (8 microM), a phospholipase C inhibitor, abolished AVP-induced increases in [Ca(2+)](i), but only reduced AVP induced glucagon release by 39%. 7. Pretreatment with nimodipine, an L-type Ca(2+) channel blocker failed to alter AVP-induced glucagon release or increase in [Ca(2+)](i). 8. The results suggest that AVP causes glucagon release through both Ca(2+)-dependent and -independent pathways. For the Ca(2+)-dependent pathway, the G(q) protein activates phospholipase C, which catalyzes the formation of IP(3). IP(3) induces Ca(2+) release from the endoplasmic reticulum, which, in turn, triggers Ca(2+) influx. Both Ca(2+) release and Ca(2+) influx may contribute to AVP-induced glucagon release. PMID- 10694232 TI - Voltage-dependent block of native AMPA receptor channels by dicationic compounds. AB - 1. The kinetics of open channel block of GluR2-containing and GluR2-lacking AMPA receptors (AMPAR) by dicationic compounds (IEM-1460, IEM-1754, and IEM-1925) have been studied in rat hippocampal neurones using whole-cell patch clamp recording and concentration-jump techniques. Neurones were isolated from hippocampal slices by vibrodissociation. 2. The dicationic compounds were approximately 100 - 200 times more potent as blockers of GluR2-lacking AMPAR than as blockers of GluR2 containing AMPAR. The subunit specificity of channel block is determined by the blocking rate constant of a dicationic compound, whereas differences in unblocking rate constants account for differences in potency. 3. Hyperpolarization may decrease the block produced by IEM-1460 and IEM-1754 block due to the voltage-dependence of the unblocking rate constants for these compounds. This suggests that dicationic compounds permeate the AMPAR channel at negative membrane potentials. The effect was particularly apparent for GluR2 lacking AMPAR. These findings indicate that the presence of GluR2-subunit(s) in AMPAR hinders the binding of the cationic compounds and their permeation through the channel. 4. The most potent compound tested was IEM-1925. The presence of a phenylcyclohexyl moiety instead of an adamantane moiety, as in IEM-1460 and IEM1754, is probably responsible for the higher potency of IEM-1925. Dicationic compounds are important not only as pharmacological tools, but also as templates for the synthesis of new selective AMPAR blockers which may be potential therapeutic agents. PMID- 10694233 TI - ADP can induce aggregation of human platelets via both P2Y(1) and P(2T) receptors. AB - 1. In the present study we have investigated the roles of P2Y(1) and P(2T) receptor subtypes in adenosine 5'-diphosphate (ADP)-induced aggregation of human platelets in heparinized platelet rich plasma. 2. The response to ADP can be characterized as the initial rate or the maximum or final extent of aggregation. The response profile is determined by the concentration of ADP used, being transient at lower and sustained at higher concentrations. 3. The P2Y(1) receptor antagonist, adenosine-3'-phosphate-5'-phosphate (A3P5P) competitively antagonized the initial rate of aggregation (pK(B) 5. 47) and transformed the response profile to a slowly developing but sustained response. Both maximum and final extents were also inhibited by A3P5P although not in a competitive manner (Schild slope <1). 4. The P(2T) receptor antagonist, AR-C67085, competitively antagonized the final extent of aggregation (pK(B) 8.54), transforming the response profile to one of rapid, transient aggregation. Its effect on maximum extent (the most widely used index of aggregation) was complex, and further supported the involvement of both receptor subtypes in the aggregation response. 5. ADP-induced aggregation is a complex phenomenon, the nature of which is determined by the relative occupancy of the two receptor subtypes. While P2Y(1) receptor activation causes a rapid and transient aggregation, the extent of sustained aggregation is determined by the level of P(2T) receptor occupancy. Hence, detailed analysis of the aggregation response is essential to correctly define the purinergic pharmacology of the platelet and interpretation of results is critically dependent on the response index chosen. PMID- 10694234 TI - Stereoselective modulatory actions of oleamide on GABA(A) receptors and voltage gated Na(+) channels in vitro: a putative endogenous ligand for depressant drug sites in CNS. AB - 1. cis-9,10-octadecenoamide ('oleamide') accumulates in CSF on sleep deprivation. It induces sleep in animals (the trans form is inactive) but its cellular actions are poorly characterized. We have used electrophysiology in cultures from embryonic rat cortex and biochemical studies in mouse nerve preparations to address these issues. 2. Twenty microM cis-oleamide (but not trans) reversibly enhanced GABA(A) currents and depressed the frequency of spontaneous excitatory and inhibitory synaptic activity in cultured networks. 3. cis-oleamide stereoselectively blocked veratridine-induced (but not K(+)-induced) depolarisation of mouse synaptoneurosomes (IC(50), 13. 9 microM). 4. The cis isomer stereoselectively blocked veratridine-induced (but not K(+)-induced) [(3)H]-GABA release from mouse synaptosomes (IC(50), 4.6 microM). 5. At 20 microM cis-oleamide, but not trans, produced a marked inhibition of Na(+) channel dependent rises in intrasynaptosomal Ca(2+). 6. The physiological significance of these observations was examined by isolating Na(+) spikes in cultured pyramidal neurones. Sixty-four microM cis-oleamide did not significantly alter the amplitude, rate of rise or duration of unitary action potentials (1 Hz). 7. cis Oleamide stereoselectively suppressed sustained repetitive firing (SRF) in these cells with an EC(50) of 4.1 microM suggesting a frequency- or state-dependent block of voltage-gated Na(+) channels. 8. Oleamide is a stereoselective modulator of both postsynaptic GABA(A) receptors and presynaptic or somatic voltage-gated Na(+) channels which are crucial for synaptic inhibition and conduction. The modulatory actions are strikingly similar to those displayed by sedative or anticonvulsant barbiturates and a variety of general anaesthetics. 9. Oleamide may represent an endogenous modulator for drug receptors and an important regulator of arousal. PMID- 10694235 TI - Effects of P(1) and P2 receptor antagonists on beta, gamma-methyleneATP- and CGS21680-induced cyclic AMP formation in NG108-15 cells. AB - 1. We have previously shown that ATP increased cyclic AMP in NG108-15 cells, which was inhibited by P(1) receptor antagonist methylxanthines. In the present study, we examined the effects of P(1) and P2 receptor antagonists on cyclic AMP formation induced by beta,gamma-methyleneATP (beta,gamma-MeATP) and CGS21680, an A(2A) adenosine receptor agonist, in NG108-15 cells. 2. beta,gamma-MeATP and CGS21680 increased intracellular cyclic AMP with EC(50) values of 8. 0+/-0.98 microM (n=4) and 42+/-7.5 nM (n=4), respectively. 3. Several P(1) receptor antagonists inhibited both beta,gamma-MeATP- and CGS21680-induced cyclic AMP increase with a similar rank order of potency; ZM241385>CGS15943>XAC>DPCPX. However, the pK(i) values of these antagonists for beta,gamma-MeATP were larger than those for CGS21680. 4. Alloxazine, a P(1) receptor antagonist, and several P2 receptor antagonists (PPADS, iPPADS, reactive blue-2) inhibited beta, gamma MeATP-induced response, while these antagonists little affected CGS21680-induced one. Suramin was effective only for beta, gamma-MeATP-induced response at 1 mM. 5. 2-chloroadenosine (2CADO) and 2-chloroATP (2ClATP) increased cyclic AMP with similar potencies. The effects of these agonists were both inhibited by ZM241385, but only 2ClATP-induced response was inhibited by PPADS. 6. ATP- and beta, gamma MeATP-induced responses were little affected by alpha, beta-methyleneADP, a 5' nucleotidase inhibitor. 7. These results clearly demonstrate that ATP-stimulated cyclic AMP formation can be distinguished from the A(2A) receptor agonist-induced one by using the several P(1) and P2 receptor antagonists. PMID- 10694236 TI - Different role of IL-4 in the onset of hapten-induced contact hypersensitivity in BALB/c and C57BL/6 mice. AB - 1. To study the role of interleukin (IL)-4 in the onset of contact hypersensitivity (CH) in mice, the effect of IL-4 gene-depletion and anti-IL-4 monoclonal antibody treatment on dinitrofluorobenzene (DNFB)-induced CH was examined. Simultaneously, to clarify the effect of background gene, DNFB-induced CH in BALB/c and C57BL/6 mice was compared. 2. Five repeated topical applications of DNFB to the ears of mice resulted in CH of the ears in terms of increases in ear thickness and histopathological changes. The magnitude of ear thickness increase in BALB/c mice was almost three times greater than that in C57BL/6 mice. 3. The CH in BALB/c mice was significantly suppressed by IL-4 gene-depletion and anti-IL-4 monoclonal antibody treatment. In contrast, the symptoms of dermatitis in C57BL/6 mice were slightly affected by the same treatment. These changes corresponded well to the production of specific IgE antibody. 4. Total IgE antibody production and the expression of productive Cepsilon mRNA were dramatically suppressed by IL-4 gene-depletion and anti-IL-4 treatment in BALB/c and C57BL/6 mice. Neither total IgG nor IgM levels in either strain of mice was altered by depletion of IL-4. 5. The expression of IFN-gamma in the skin lesion was dramatically suppressed by IL-4 gene-depletion in BALB/c mice, but not in C57BL/6 mice. 6. These findings indicate that IL-4 plays an important role in the onset of DNFB-induced CH in BALB/c mice, but not in C57BL/6 mice. PMID- 10694237 TI - Lipopolysaccharide activates nuclear factor kappaB in rat intestine: role of endogenous platelet-activating factor and tumour necrosis factor. AB - 1. We examined the effect of lipopolysaccharide (LPS), a cell wall constituent of Gram negative bacteria, on nuclear factor kappaB (NF-kappaB) activation in the intestine and the roles of endogenous platelet-activating factor (PAF), tumour necrosis factor-alpha (TNF) and neutrophils. We also compared the time course of NF-kappaB activation in response to PAF and LPS. 2. Ileal nuclear extracts from LPS (8 mg kg(-1), IV)-injected rats were assayed for NF-kappaB-DNA-binding activity and identification of the subunits. Some rats were pretreated with WEB2170 (a PAF receptor antagonist), anti-TNF antibody, or anti-neutrophil antiserum. NF-kappaB p65 was localized by immunohistochemistry. An additional group was challenged with PAF (2 microg kg(-1), IV) for comparison. 3. LPS activates intestinal NF-kappaB, both as p50-p50 and p50-p65 dimers within 15 min, and the effect peaks at 2 h. The effect is slower and more sustained than that of PAF, which peaks at 30 min. Activated NF-kappaB was immunolocalized within epithelial and lamina propria cells. LPS effect was reduced by 41, 37 and 44%, respectively, in animals pretreated with WEB2170, anti-TNF antibody, or anti neutrophil antiserum (P<0.05). 4. LPS activates intestinal NF-kappaB in vivo and neutrophil activation is involved in its action. The LPS effect is mediated by both endogenous PAF and TNF. PMID- 10694238 TI - Differential actions of L-cysteine on responses to nitric oxide, nitroxyl anions and EDRF in the rat aorta. AB - 1. The effects of L-cysteine were tested in rat aortic rings on responses to nitric oxide free radical (NO(*)), nitroxyl (NO(-)) derived from Angeli's salt and endothelium-derived relaxing factor (EDRF) activated by acetylcholine, ATP and the calcium ionophore A23187. Concentrations of 300 microM or less of L cysteine had no effect on responses. 2. Relaxations produced by exogenous NO(*) (0.25 - 2.5 microM) were markedly prolonged and relaxations produced by sodium nitroprusside (0.001 - 0.3 microM) were enhanced by 1 and 3 mM L-cysteine. The enhancements by L-cysteine of responses to NO(*) and sodium nitroprusside may be attributed to the formation of S-nitrosocysteine. 3. Relaxations mediated by the nitroxyl anion (0.3 microM) donated from Angeli's salt were more prolonged than those produced by NO(*), and nitroxyl-induced relaxations were reduced by L cysteine (1 and 3 mM). 4. EDRF-mediated relaxations produced by acetylcholine (0.01 - 10 microM), ATP (3 - 100 microM) and the calcium ionophore A23187 (0.1 microM) were significantly reduced by 3 mM L-cysteine. 5. The similarity between the inhibitory effects of L-cystei on responses to EDRF and on those to nitroxyl suggests that a component of the response to EDRF may be mediated by nitroxyl anion. PMID- 10694239 TI - Effects of mefloquine on cardiac contractility and electrical activity in vivo, in isolated cardiac preparations, and in single ventricular myocytes. AB - 1. To examine the possible cardiotoxicity of the antimalarial drug mefloquine, increasing doses (0.3 - 30 mg kg(-1)) were given i.v. to anaesthetized guinea pigs. Mefloquine did not alter ECG intervals significantly but gradually increased systolic blood pressure (at 3 mg kg(-1)) then had a depressor effect (at 10 mg kg(-1)). Death due to profound hypotension, probably resulting from cardiac contractile failure or AV block, occurred after either 10 mg kg(-1) (2/6) or 30 mg kg(-1) (4/6) mefloquine. 2. In isolated cardiac preparations mefloquine (3 - 100 microM) did not alter the effective refractory period but at the higher concentrations resting tension increased. Developed tension was reduced by 100 microM mefloquine in left atria (from 5.8+/-1.7 to 2.2+/-0.4 mN) whereas in papillary muscles although 30 microM mefloquine reduced developed tension (from 2. 6+/-0.5 to 1.1+/-0.1 mN) subsequent addition of 100 microM caused a marked, but not sustained, positive inotropic effect (from 1.2+/-0.1 to 3.8+/-0.8 mN). 3. In single ventricular myocytes, mefloquine (10 microM) shortened action potential duration (e.g. APD(90) from 285+/-29 to 141+/-12 ms) and reduced the amplitude of the systolic Ca(2+) transient. 4. These effects were accompanied by a decrease in the L-type Ca(2+) current. These results indicate that the main adverse effect of mefloquine on the heart is a negative inotropic action. This action can be explained by blockade of L-type Ca(2+) channels. PMID- 10694240 TI - Primary porcine enterocyte and hepatocyte cultures to study drug oxidation reactions. AB - 1. Primary porcine hepatocytes and enterocytes were isolated and cultured in Williams' E medium for up to 10 days to investigate potential organ differences in the metabolism of the immunosuppressive compound tacrolimus (FK 506) and of two investigational drugs (KC11346 and KC12291). Using LC-MS (FK506) and HPLC-FL (KC 11346/12291) a number of metabolites with identical mass and/or identical retention time could be detected. 2. In the case of tacrolimus hepatocytes and enterocytes produced the same spectrum of metabolites, e.g. bisdemethyl tacrolimus, demethyl-tacrolimus, demethyl-hydroxy-tacrolimus and hydroxy tacrolimus, albeit at varying intensities. 3. Treatment of enterocyte cultures with dexamethasone increased the overall metabolite formation very significantly (up to 36 fold). 4. The metabolism of tacrolimus was also studied with preparations of insect cells, that express specifically high levels of individual human cytochrome P450 (CYP) isoenzymes. All metabolites could be generated with microsomal preparations specifically expressing CYP3A4, but hydroxy-tacrolimus was exclusively produced by CYP3A5. 5. In the case of the investigational drugs KC 11346 and KC 12291 only three metabolites were formed by cultured enterocytes whereas hepatocytes produced 10 and 20 metabolites, respectively. 6. When assessed at the protein level CYP1A and CYP3A were expressed in cultures of porcine enterocytes for up to 10 days but porcine hepatocytes expressed additionally CYP2C9/10. 7. In conclusion, primary enterocytes and hepatocytes can be successfully cultured for several days while maintaining mono-oxygenase activity and may therefore be used as a tool for studying intestinal and hepatic metabolism. PMID- 10694241 TI - A comparison of the anti-inflammatory and anti-nociceptive activity of nitroaspirin and aspirin. AB - 1. Nitroaspirin (2.5 - 50 mg kg(-1), i.p. or 2.5 - 100 mg kg(-1), p.o.) and aspirin (2.5 - 100 mg kg(-1), i.p. or p.o.) exhibit anti-inflammatory activity in the carrageenan-induced hindpaw oedema model in the rat. When administered i.p., nitroaspirin was a more effective anti-oedema agent than aspirin particularly in the 'early' phase (i.e. up to 60 min) of the response. The ED(50) values for nitroaspirin and aspirin as inhibitors of the 'late' phase response (measured at 180 min) were 64.3 micromol kg(-1) and >555 micromol kg(-1), respectively. When administered p.o., neither nitroaspirin nor aspirin exhibited significant anti inflammatory activity in the 'early' phase and were of similar potency in the 'late' phase. Thus, at the highest dose used (100 mg kg(-1), 360 min) orally administered nitroaspirin (aspirin in parenthesis) inhibited oedema formation by 46.9+/-1.6% (47.2+/-3.8%, both n=6, P<0.05). 2. Nitroaspirin and aspirin (25 - 200 mg kg(-1), p.o.) caused dose-related inhibition of the hyperalgesia to mechanical stimulation following intraplantar injection of carrageenan in the rat. ED(50) values were 365 micromol kg(-1) and 784 micromol kg(-1), respectively. Neither drug influenced the threshold for mechanical stimulation in the contralateral (i.e. untreated) hindpaw. 3. Nitroaspirin and aspirin (2.5 - 100 mg kg(-1), p.o.) caused dose-related inhibition of acetic acid induced abdominal constrictions in the mouse (ED(50) values of 154.7 micromol kg(-1) and 242.8 micromol kg(-1), respectively). 4. Nitroaspirin and aspirin (>200 mg kg( 1), p.o.) reduced the 'late' phase (but not the 'early' phase) of the formalin induced hindpaw licking assay in the mouse. Similarly, nitroaspirin and aspirin (>50 mg kg(-1), p.o.) prolonged tail withdrawal latency following application of a noxious heat stimulus in the mouse. PMID- 10694242 TI - Effects of spinally administered P2X receptor agonists and antagonists on the responses of dorsal horn neurones recorded in normal, carrageenan-inflamed and neuropathic rats. AB - 1. The function and role of P2X receptors in the spinal transmission of nociception was investigated using the selective P2X receptor agonists, alpha,beta-methylene ATP (alpha,beta-me ATP) and beta, gamma-methylene-L-ATP (beta,gamma-me-L-ATP) and the P2X receptor antagonists pyridoxal-phosphate-6 azophenyl-2',4'-disulphonate (PPADS) and suramin. 2. Intrathecal administration of 5 and 50 microg of beta,gamma-me-L-ATP produced a significant facilitation of the C-fibre evoked response and a tendency towards increased excitability of the post-discharge, but not Abeta-fibre evoked response of dorsal horn neurones recorded in normal animals. Administration of similar doses of alpha,beta-me ATP did not produce an overall change in the response of the neuronal population. 3. Peripheral administration of 20 microg of these agonists into the paw of the rat evoked firing in the dorsal horn neurones. 4. Intrathecal administration of the antagonists, suramin (50 and 500 microg) and PPADS (5, 50 and 500 microg), to normal animals and to animals with a model of neuropathy induced by spinal nerve ligation did not alter the evoked neuronal responses. In contrast, intrathecal administration of 500 microg of suramin to animals 3 h after the induction of carrageenan inflammation produced a significant inhibition of the C-fibre evoked response of the neurones. Similar inhibitions were also seen following high doses of intrathecal PPADS, although this did not reach significance. 5. These results suggest that spinal P2X receptors may play a role in the modulation of spinal nociceptive transmission following the development of inflammation, but that these receptors play at most a minor role in spinal nociceptive processing in normal and neuropathic animals. PMID- 10694243 TI - Antagonism of calcium currents and neurotransmitter release by barium ions at frog motor nerve endings. AB - 1. The effects of Ba(2+) (0.1 - 2 mM) on the component of the perineural voltage change associated with nerve terminal calcium currents (prejunctional Ca(2+) currents) were compared with the effects of this ion to antagonize calcium dependent acetylcholine (ACh) release. These experiments were made on isolated neuromuscular junctions of the frog. 2. In the presence of sufficient concentrations of K(+) channel blockers to eliminate measurable prejunctional K(+) currents, low concentrations of Ba(2+) selectively antagonized prejunctional Ca(2+) currents in normal Ca(2+) solutions. Higher concentrations of Ba(2+) also substantially reduced the Na(+) component of the perineural waveform. 3. Ba(2+) inhibited the prolonged prejunctional Ca(2+) currents that developed in the presence of higher concentrations of K(+) channel blockers. 4. Simultaneous measurements of the prejunctional Ca(2+) currents and the electrophysiological correlates of ACh release (i.e. end-plate potentials, EPPs) were made under conditions of modest K(+) channel blockade. Under these conditions, Ba(2+) generally produced simultaneous decreases in both Ca(2+) currents and EPP amplitudes. In some instances, a prolongation of prejunctional Ca(2+) currents and a transient increase in EPP amplitudes preceded the decreases in both electrophysiological events. 5. These results suggest that Ba(2+) ions can antagonize the entry of calcium into motor nerve endings and this effect is likely to be responsible for the inhibitory effects of Ba(2+) on evoked ACh release. PMID- 10694244 TI - Inhibition by troglitazone of the antigen-induced production of leukotrienes in immunoglobulin E-sensitized RBL-2H3 cells. AB - 1. The effect of troglitazone, an anti-diabetic drug with insulin-sensitizing action, on antigen-induced production of leukotriene (LT) B(4), C(4) and E(4) and prostaglandin D(2) (PGD(2)) was examined in dinitrophenol (DNP)-specific immunoglobulin E (IgE)-sensitized RBL-2H3 mast cells following stimulation by the antigen, DNP-conjugated human serum albumin. Levels of LTB(4), C(4) and E(4) and PGD(2) in the conditioned medium were enzyme-immunoassayed. 2. Troglitazone inhibited the antigen-induced production of LTB(4), C(4) and E(4) and the potency of the inhibition was comparable to that of zileuton, a specific inhibitor of 5 lipoxygenase (5-LOX) and a clinically used anti-asthmatic drug. Neither troglitazone nor zileuton affected antigen-induced production of PGD(2), arachidonic acid release from membrane phospholipids and degranulation. 3. Troglitazone inhibited LTB(4) production by the supernatant fraction of RBL-2H3 cell lysate with similar potency to zileuton, suggesting that troglitazone inhibits LT production by direct inhibition of 5-LOX activity. 4. Furthermore, it was shown that troglitazone as well as zileuton inhibited LTB(4) production in A23187-stimulated rat peritoneal neutrophils. 5. These findings suggest that troglitazone inhibits antigen-induced LT production in the IgE-sensitized RBL-2H3 cells and A23187-stimulated rat peritoneal neutrophils by direct inhibition of 5 LOX activity. PMID- 10694245 TI - Tonic activation of A(2A) adenosine receptors unmasks, and of A(1) receptors prevents, a facilitatory action of calcitonin gene-related peptide in the rat hippocampus. AB - 1. We investigated how manipulations of the degree of activation of adenosine A(1) and A(2A) receptors influences the action of the neuropeptide, calcitonin gene-related peptide (CGRP) on synaptic transmission in hippocampal slices. Field excitatory post-synaptic potentials (EPSPs) from the CA1 area were recorded. 2. When applied alone, CGRP (1 - 30 nM) was without effect on field EPSPs. However, CGRP (10 - 30 nM) significantly increased the field EPSP slope when applied to hippocampal slices in the presence of the A(1) receptor antagonist, 1,3-dipropyl 8-cyclopenthyl xanthine (DPCPX, 10 nM), or in the presence of the A(2A) adenosine receptor agonist CGS 21680 (10 nM). 3. The A(2A) receptor antagonist, ZM 241385 (10 nM) as well as adenosine deaminase (ADA, 2 U ml(-1)), prevented the enhancement of field EPSP slope caused by CGRP (30 nM) in the presence of DPCPX (10 nM), suggesting that this effect of CGRP requires the concomitant activation of A(2A) adenosine receptors by endogenous adenosine. 4. The protein kinase-A inhibitors, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA-1004, 10 microM) and adenosine 3',5'-cyclic monophosphorothioate, Rp-isomer (Rp-cAMPS, 50 microM), as well as the inhibitor of ATP-sensitive potassium (K(ATP)) channels, glibenclamide (30 microM), prevented the facilitation of synaptic transmission caused by CGRP (30 nM) in the presence of DPCPX (10 nM), suggesting that this effect of CGRP involves both K(ATP) channels and protein kinase-A. 5. It is concluded that the ability of CGRP to facilitate synaptic transmission in the CA1 area of the hippocampus is under tight control by adenosine, with tonic A(1) receptor activation by endogenous adenosine 'braking' the action of CGRP, and the A(2A) receptors triggering this action. PMID- 10694246 TI - An indirect influence of phenylephrine on the release of endothelium-derived vasodilators in rat small mesenteric artery. AB - 1. The possibility that stimulation of smooth muscle alpha(1)-adrenoceptors modulates contraction via the endothelium was examined in rat small mesenteric arteries. 2. N(omega)-nitro-L-arginine methyl ester, (L-NAME, 100 microM to inhibit NO synthase) increased contraction to single concentrations of phenylephrine (1 - 3 microM) by approximately 2 fold (from a control level of 14.2+/-3.0 to 34. 1+/-4.2% of the maximum contraction of the artery, n=20). The action of L-NAME was abolished by disrupting the endothelium. 3. The subsequent addition of apamin (to inhibit small conductance Ca(2+)-activated K(+) channels, 50 nM) further augmented phenylephrine contractions, in an endothelium-dependent manner, to more than 3 fold above control (50.4+/-5.3% of the maximum contraction, n=11). 4.Charybdotoxin (non-selective inhibitor of large conductance Ca(2+)-activated K(+) channels, BK(Ca), 50 nM) plus L-NAME augmented the level of phenylephrine contraction to 4 - 5-fold above control (64.1+/-3.1%, n=5), but this effect was independent of the endothelium. The potentiation of contraction by charybdotoxin could be mimicked with the selective BK(Ca) inhibitor, iberiotoxin,. 5. Apamin together with L-NAME and charybdotoxin further significantly increased the phenylephrine contraction by 5 - 6-fold, to 79.9+/ 3.5% of the maximum contraction of the artery (n=13). 6. Phenylephrine failed directly to increase the intracellular Ca(2+) concentration in endothelial cells freshly isolated from the small mesenteric artery. 7. Stimulation of smooth muscle alpha(1)-adrenoceptors in the mesenteric artery induces contraction that is markedly suppressed by the endothelium. The attenuation of contraction appears to reflect both the release of NO from the endothelium and the efflux of K(+) from both endothelial and smooth muscle cells. This suggests that the release of NO and endothelium-derived hyperpolarizing factor can be evoked indirectly by agents which act only on the smooth muscle cells. PMID- 10694247 TI - Functional characterization of the P2X(4) receptor orthologues. AB - 1. The aim of this study was to functionally characterize the recombinant mouse P2X(4) receptor and to compare its pharmacological properties with those of the human and rat orthologues. 2. Whole cell recordings were made from rafts of HEK 293 cells stably expressing recombinant mouse, rat or human P2X(4) receptors, using Cs-aspartate containing electrodes (3 - 8 MOmega) in a HEPES-buffered extracellular medium. 3. The agonist potency of ATP at the three species orthologues was similar, with mean EC(50) values of 2.3 microM, 1.4 microM and 5.5 microM, respectively. 4. Adenosine-5'-tetraphosphate (AP4) acted as a partial agonist with respect to ATP at the mouse and human P2X(4) receptors (EC(50)=2.6 and 3.0 microM), but was significantly less potent at the rat orthologue (EC(50)=20.0 microM). alpha,beta-methylene adenosine-5'-triphosphate (alpha,beta meATP) also acted as a partial agonist, producing 29% of the maximum response at the mouse P2X(4) and 24% at the human P2X(4) receptor. 5. In contrast to the other species orthologues, alpha,beta-meATP failed to elicit a significant agonist response at rat P2X(4) receptors, and was found to act as an antagonist, with an IC(50) of 4.6 microM, against 10 microM ATP. 6. Mouse P2X(4) receptors were found to be sensitive to the antagonist, pyridoxalphosphate-6-azophenyl 2',4'-disulphonic acid (PPADS) (IC(50)=10.5 microM), as were human P2X(4) receptors (IC(50)=9.6 microM). The rat receptor however, showed a low sensitivity to PPADS (IC(50)>100 microM). 7. All three orthologues were relatively suramin insensitive (IC(50)>100 microM) and insensitive to 1-[N, O-Bis(5-isoquinoline sulphonyl)benzyl]-2-(4-phenylpiperazine)ethyl]-5-isoquinoline sulphonamide (KN 62; IC(50)>3 microM). 8. Our results suggest that the pharmacological properties of the mouse receptor are most similar to the human P2X(4) receptor, and differ markedly from the rat receptor. PMID- 10694248 TI - Impaired relaxation of stomach smooth muscle in mice lacking cyclic GMP-dependent protein kinase I. AB - 1. Guanosine 3', 5'-cyclic monophosphate (cyclic GMP)-dependent kinase I (cGKI) is a major receptor for cyclic GMP in a variety of cells. Mice lacking cGKI exhibit multiple phenotypes, including severe defects in smooth muscle function. We have investigated the NO/cGMP- and vasoactive intestinal polypeptide (VIP)/adenosine 3', 5'-cyclic monophosphate (cyclic AMP)-signalling pathways in the gastric fundus of wild type and cGKI-deficient mice. 2. Using immunohistochemistry, similar staining patterns for NO-synthase, cyclic GMP- and VIP-immunoreactivities were found in wild type and cGKI-deficient mice. 3. In isolated, endothelin-1 (3 nM - 3 microM)-contracted, muscle strips from wild type mice, electrical field stimulation (1 - 16 Hz) caused a biphasic relaxation, one initial rapid, followed by a more slowly developing phase. In preparations from cGKI-deficient mice only the slowly developing relaxation was observed. 4. The responses to the NO donor, SIN-1 (10 nM - 100 microM), and to 8-Br-cyclic GMP (10 nM - 100 microM) were markedly impaired in strips from cGKI-deficient mice, whereas the responses to VIP (0.1 nM - 1 microM) and forskolin (0.1 nM - 1 microM) were similar to those in wild type mice. 5. These results suggest that cGKI plays a central role in the NO/cGMP signalling cascade producing relaxation of mouse gastric fundus smooth muscle. Relaxant agents acting via the cyclic AMP pathway can exert their effects independently of cGKI. PMID- 10694249 TI - Modulation by bicuculline and penicillin of the block by t-butyl-bicyclo phosphorothionate (TBPS) of GABA(A)-receptor mediated Cl(-)-current responses in rat striatal neurones. AB - 1. T-butyl-bicyclo-phosphorothionate (TBPS) is a prototypical representative of the cage-convulsants which act through a use-dependent block of the GABA(A) receptor-ionophore complex. Using current recordings from cultured neurones of rat striatum the manner was investigated in which two antagonists, bicuculline and penicillin, presumably acting at the agonist binding site and in the ionic channel, respectively, modify the rate of block by TBPS. 2. Penicillin (5 or 10 mM) did not slow the rate of block by TBPS, but produced a significant enhancement of block rate, which, however, was inversely related to the degree of antagonism by penicillin of the GABA-induced current. 3. Bicuculline (10 microM) reduced the rate of block by TBPS. However, this effect was 3 fold weaker than its GABA-antagonistic action. The slowing of block rate and the current antagonism exhibited a biphasic, positive-negative relationship. Co-application of bicuculline (100 microM) in a concentration that produced nearly complete antagonism and TBPS (10 microM) resulted in a marked ( approximately 40%) reduction of subsequent GABA response amplitudes compatible with a direct, bicuculline-induced conformational change in the receptor required for the binding of and block by TBPS. 4. The lack of protection afforded by the channel blocker penicillin as well as the lack of correlation between bicuculline antagonism of the Cl(-)-current and its efficiency in protecting against TBPS block is evidence against an open channel blocking mechanism for TBPS. TBPS does, therefore, not appear to gain access to its binding site via the open pore but through alternative routes regulated from the agonist binding site. PMID- 10694250 TI - Quantification of state-dependent drug interactions with the sodium channel. PMID- 10694251 TI - Reply PMID- 10694252 TI - Trapping of a methanesulfonanilide by closure of the HERG potassium channel activation gate. AB - Deactivation of voltage-gated potassium (K(+)) channels can slow or prevent the recovery from block by charged organic compounds, a phenomenon attributed to trapping of the compound within the inner vestibule by closure of the activation gate. Unbinding and exit from the channel vestibule of a positively charged organic compound should be favored by membrane hyperpolarization if not impeded by the closed gate. MK-499, a methanesulfonanilide compound, is a potent blocker (IC(50) = 32 nM) of HERG K(+) channels. This bulky compound (7 x 20 A) is positively charged at physiological pH. Recovery from block of HERG channels by MK-499 and other methanesulfonanilides is extremely slow (Carmeliet 1992; Ficker et al. 1998), suggesting a trapping mechanism. We used a mutant HERG (D540K) channel expressed in Xenopus oocytes to test the trapping hypothesis. D540K HERG has the unusual property of opening in response to hyperpolarization, in addition to relatively normal gating and channel opening in response to depolarization (Sanguinetti and Xu 1999). The hyperpolarization-activated state of HERG was characterized by long bursts of single channel reopening. Channel reopening allowed recovery from block by 2 microM MK-499 to occur with time constants of 10.5 and 52.7 s at -160 mV. In contrast, wild-type HERG channels opened only briefly after membrane hyperpolarization, and thus did not permit recovery from block by MK-499. These findings provide direct evidence that the mechanism of slow recovery from HERG channel block by methanesulfonanilides is due to trapping of the compound in the inner vestibule by closure of the activation gate. The ability of HERG channels to trap MK-499, despite its large size, suggests that the vestibule of this channel is larger than the well studied Shaker K(+) channel. PMID- 10694253 TI - Single-channel properties in endoplasmic reticulum membrane of recombinant type 3 inositol trisphosphate receptor. AB - The inositol 1,4,5-trisphosphate receptor (InsP(3)R) is an intracellular Ca(2+) release channel localized in endoplasmic reticulum (ER) with a central role in complex Ca(2+) signaling in most cell types. A family of InsP(3)Rs encoded by several genes has been identified with different primary sequences, subcellular locations, variable ratios of expression, and heteromultimer formation. This diversity suggests that cells require distinct InsP(3)Rs, but the functional correlates of this diversity are largely unknown. Lacking are single-channel recordings of the recombinant type 3 receptor (InsP(3)R-3), a widely expressed isoform also implicated in plasma membrane Ca(2+) influx and apoptosis. Here, we describe functional expression and single-channel recording of recombinant rat InsP(3)R-3 in its native membrane environment. The approach we describe suggests a novel strategy for expression and recording of recombinant ER-localized ion channels in the ER membrane. Ion permeation and channel gating properties of the rat InsP(3)R-3 are strikingly similar to those of Xenopus type 1 InsP(3)R in the same membrane. Using two different two-electrode voltage clamp protocols to examine calcium store-operated calcium influx, no difference in the magnitude of calcium influx was observed in oocytes injected with rat InsP(3)R-3 cRNA compared with control oocytes. Our results suggest that if cellular expression of multiple InsP(3)R isoforms is a mechanism to modify the temporal and spatial features of [Ca(2+)](i) signals, then it must be achieved by isoform-specific regulation or localization of various types of InsP(3)Rs that have relatively similar Ca(2+) permeation properties. PMID- 10694254 TI - Independence and cooperativity in rearrangements of a potassium channel voltage sensor revealed by single subunit fluorescence. AB - Voltage-gated potassium channels are composed of four subunits. Voltage-dependent activation of these channels consists of a depolarization-triggered series of charge-carrying steps that occur in each subunit. These major charge-carrying steps are followed by cooperative step(s) that lead to channel opening. Unlike the late cooperative steps, the major charge-carrying steps have been proposed to occur independently in each of the channel subunits. In this paper, we examine this further. We showed earlier that the two major charge-carrying steps are associated with two sequential outward transmembrane movements of the charged S4 segment. We now use voltage clamp fluorometry to monitor these S4 movements in individual subunits of heterotetrameric channels. In this way, we estimate the influence of one subunit's S4 movement on another's when the energetics of their transmembrane movements differ. Our results show that the first S4 movement occurs independently in each subunit, while the second occurs cooperatively. At least part of the cooperativity appears to be intrinsic to the second S4 charge carrying rearrangement. Such cooperativity in gating of voltage-dependent channels has great physiological relevance since it can affect both action potential threshold and rate of propagation. PMID- 10694255 TI - The barium site in a potassium channel by x-ray crystallography. AB - X-ray diffraction data were collected from frozen crystals (100 degrees K) of the KcsA K(+) channel equilibrated with solutions containing barium chloride. Difference electron density maps (F(barium) - F(native), 5.0 A resolution) show that Ba(2+) resides at a single location within the selectivity filter. The Ba(2+) blocking site corresponds to the internal aspect (adjacent to the central cavity) of the "inner ion" position where an alkali metal cation is found in the absence of the blocking Ba(2+) ion. The location of Ba(2+) with respect to Rb(+) ions in the pore is in good agreement with the findings on the functional interaction of Ba(2+) with K(+) (and Rb(+)) in Ca(2+)-activated K(+) channels (Neyton, J., and C. Miller. 1988. J. Gen. Physiol. 92:549-567). Taken together, these structural and functional data imply that at physiological ion concentrations a third ion may interact with two ions in the selectivity filter, perhaps by entering from one side and displacing an ion on the opposite side. PMID- 10694256 TI - Modulating modulation. PMID- 10694257 TI - Modulation of N-type calcium channel activity by G-proteins and protein kinase C. AB - N-type voltage-gated calcium channel activity in rat superior cervical ganglion neurons is modulated by a variety of pathways. Activation of heterotrimeric G proteins reduces whole-cell current amplitude, whereas phosphorylation by protein kinase C leads to an increase in current amplitude. It has been proposed that these two distinct pathways converge on the channel's pore-forming alpha(1B) subunit, such that the actions of one pathway can preclude those of the other. In this study, we have characterized further the actions of PKC on whole-cell barium currents in neonatal rat superior cervical ganglion neurons. We first examined whether the effects of G-protein-mediated inhibition and phosphorylation by PKC are mutually exclusive. G-proteins were activated by including 0.4 mM GTP or 0.1 mM GTP-gamma-S in the pipette, and PKC was activated by bath application of 500 nM phorbol 12-myristate 13-acetate (PMA). We found that activated PKC was unable to reverse GTP-gamma-S-induced inhibition unless prepulses were applied, indicating that reversal of inhibition by phosphorylation appears to occur only after dissociation of the G-protein from the channel. Once inhibition was relieved, activation of PKC was sufficient to prevent reinhibition of current by G-proteins, indicating that under phosphorylating conditions, channels are resistant to G-protein-mediated modulation. We then examined what effect, if any, phosphorylation by PKC has on N-type barium currents beyond antagonizing G protein-mediated inhibition. We found that, although G-protein activation significantly affected peak current amplitude, fast inactivation, holding potential-dependent inactivation, and voltage-dependent activation, when G protein activation was minimized by dialysis of the cytoplasm with 0.1 mM GDP beta-S, these parameters were not affected by bath application of PMA. These results indicate that, under our recording conditions, phosphorylation by PKC has no effect on whole-cell N-type currents, other than preventing inhibition by G proteins. PMID- 10694258 TI - Cytoplasmic unsaturated free fatty acids inhibit ATP-dependent gating of the G protein-gated K(+) channel. AB - This study reports the identification of an endogenous inhibitor of the G protein gated (K(ACh)) channel and its effect on the K(ACh) channel kinetics. In the presence of acetylcholine in the pipette, K(ACh) channels in inside-out atrial patches were activated by applying GTP to the cytoplasmic side of the membrane. In these patches, addition of physiological concentration of intracellular ATP (4 mM) upregulated K(ACh) channel activity approximately fivefold and induced long lived openings. However, such ATP-dependent gating is normally not observed in cell-attached patches, indicating that an endogenous substance that inhibits the ATP effect is present in the cell. We searched for such an inhibitor in the cell. ATP-dependent gating of the K(ACh) channel was inhibited by the addition of the cytosolic fraction of rat atrial or brain tissues. The lipid component of the cytosolic fraction was found to contain the inhibitory activity. To identify the lipid inhibitor, we tested the effect of approximately 40 different lipid molecules. Among the lipids tested, only unsaturated free fatty acids such as oleic, linoleic, and arachidonic acids (0.2-2 microM) reversibly inhibited the ATP-dependent gating of native K(ACh) channels in atrial cells and hippocampal neurons, and of recombinant K(ACh) channels (GIRK1/4 and GIRK1/2) expressed in oocytes. Unsaturated free fatty acids also inhibited phosphatidylinositol-4, 5 bisphosphate (PIP(2))-induced changes in K(ACh) channel kinetics but were ineffective against ATP-activated background K(1) channels and PIP(2)-activated K(ATP) channels. These results show that during agonist-induced activation, unsaturated free fatty acids in the cytoplasm help to keep the cardiac and neuronal K(ACh) channels downregulated by antagonizing their ATP-dependent gating. The opposing effects of ATP and free fatty acids represent a novel regulatory mechanism for the G protein-gated K(+) channel. PMID- 10694259 TI - Activation of Drosophila sodium channels promotes modification by deltamethrin. Reductions in affinity caused by knock-down resistance mutations. AB - kdr and super-kdr are mutations in houseflies and other insects that confer 30- and 500-fold resistance to the pyrethroid deltamethrin. They correspond to single (L1014F) and double (L1014F+M918T) mutations in segment IIS6 and linker II(S4-S5) of Na channels. We expressed Drosophila para Na channels with and without these mutations and characterized their modification by deltamethrin. All wild-type channels can be modified by <10 nM deltamethrin, but high affinity binding requires channel opening: (a) modification is promoted more by trains of brief depolarizations than by a single long depolarization, (b) the voltage dependence of modification parallels that of channel opening, and (c) modification is promoted by toxin II from Anemonia sulcata, which slows inactivation. The mutations reduce channel opening by enhancing closed-state inactivation. In addition, these mutations reduce the affinity for open channels by 20- and 100 fold, respectively. Deltamethrin inhibits channel closing and the mutations reduce the time that channels remain open once drug has bound. The super-kdr mutations effectively reduce the number of deltamethrin binding sites per channel from two to one. Thus, the mutations reduce both the potency and efficacy of insecticide action. PMID- 10694260 TI - Extracellular Mg(2+) modulates slow gating transitions and the opening of Drosophila ether-a-Go-Go potassium channels. AB - We have characterized the effects of prepulse hyperpolarization and extracellular Mg(2+) on the ionic and gating currents of the Drosophila ether-a-go-go K(+) channel (eag). Hyperpolarizing prepulses significantly slowed channel opening elicited by a subsequent depolarization, revealing rate-limiting transitions for activation of the ionic currents. Extracellular Mg(2+) dramatically slowed activation of eag ionic currents evoked with or without prepulse hyperpolarization and regulated the kinetics of channel opening from a nearby closed state(s). These results suggest that Mg(2+) modulates voltage-dependent gating and pore opening in eag channels. To investigate the mechanism of this modulation, eag gating currents were recorded using the cut-open oocyte voltage clamp. Prepulse hyperpolarization and extracellular Mg(2+) slowed the time course of ON gating currents. These kinetic changes resembled the results at the ionic current level, but were much smaller in magnitude, suggesting that prepulse hyperpolarization and Mg(2+) modulate gating transitions that occur slowly and/or move relatively little gating charge. To determine whether quantitatively different effects on ionic and gating currents could be obtained from a sequential activation pathway, computer simulations were performed. Simulations using a sequential model for activation reproduced the key features of eag ionic and gating currents and their modulation by prepulse hyperpolarization and extracellular Mg(2+). We have also identified mutations in the S3-S4 loop that modify or eliminate the regulation of eag gating by prepulse hyperpolarization and Mg(2+), indicating an important role for this region in the voltage-dependent activation of eag. PMID- 10694261 TI - Impaired calcium release in cerebellar Purkinje neurons maintained in culture. AB - Cerebellar Purkinje neurons demonstrate a form of synaptic plasticity that, in acutely prepared brain slices, has been shown to require calcium release from the intracellular calcium stores through inositol trisphosphate (InsP(3)) receptors. Similar studies performed in cultured Purkinje cells, however, find little evidence for the involvement of InsP(3) receptors. To address this discrepancy, the properties of InsP(3)- and caffeine-evoked calcium release in cultured Purkinje cells were directly examined. Photorelease of InsP(3) (up to 100 microM) from its photolabile caged analogue produced no change in calcium levels in 70% of cultured Purkinje cells. In the few cells where a calcium increase was detected, the response was very small and slow to peak. In contrast, the same concentration of InsP(3) resulted in large and rapidly rising calcium responses in all acutely dissociated Purkinje cells tested. Similar to InsP(3), caffeine also had little effect on calcium levels in cultured Purkinje cells, yet evoked large calcium transients in all acutely dissociated Purkinje cells tested. The results demonstrate that calcium release from intracellular calcium stores is severely impaired in Purkinje cells when they are maintained in culture. Our findings suggest that cultured Purkinje cells are an unfaithful experimental model for the study of the role of calcium release in the induction of cerebellar long term depression. PMID- 10694262 TI - Mitochondrial memory banks. Calcium stores keep a record of neuronal stimulation. PMID- 10694263 TI - Dissection of mitochondrial Ca2+ uptake and release fluxes in situ after depolarization-evoked [Ca2+](i) elevations in sympathetic neurons. AB - We studied how mitochondrial Ca2+ transport influences [Ca2+](i) dynamics in sympathetic neurons. Cells were treated with thapsigargin to inhibit Ca2+ accumulation by SERCA pumps and depolarized to elevate [Ca2+(i); the recovery that followed repolarization was then examined. The total Ca2+ flux responsible for the [Ca2+](i) recovery was separated into mitochondrial and nonmitochondrial components based on sensitivity to the proton ionophore FCCP, a selective inhibitor of mitochondrial Ca2+ transport in these cells. The nonmitochondrial flux, representing net Ca2+ extrusion across the plasma membrane, has a simple dependence on [Ca2+](i), while the net mitochondrial flux (J(mito)) is biphasic, indicative of Ca+) accumulation during the initial phase of recovery when [Ca2+](i) is high, and net Ca2+ release during later phases of recovery. During each phase, mitochondrial Ca2+ transport has distinct effects on recovery kinetics. J(mito) was separated into components representing mitochondrial Ca2+ uptake and release based on sensitivity to the specific mitochondrial Na(+)/Ca2+ exchange inhibitor, CGP 37157 (CGP). The CGP-resistant (uptake) component of J(mito) increases steeply with [Ca2+](i), as expected for transport by the mitochondrial uniporter. The CGP-sensitive (release) component is inhibited by lowering the intracellular Na(+) concentration and depends on both intra- and extramitochondrial Ca2+ concentration, as expected for the Na(+)/Ca2+ exchanger. Above approximately 400 nM [Ca2+](i), net mitochondrial Ca2+ transport is dominated by uptake and is largely insensitive to CGP. When [Ca2+](i) is approximately 200-300 nM, the net mitochondrial flux is small but represents the sum of much larger uptake and release fluxes that largely cancel. Thus, mitochondrial Ca2+ transport occurs in situ at much lower concentrations than previously thought, and may provide a mechanism for quantitative control of ATP production after brief or low frequency stimuli that raise [Ca(2+)](i) to levels below approximately 500 nM. PMID- 10694265 TI - Preimplantation genetic diagnosis of a reciprocal translocation t(3;11)(q27.3;q24.3) in siblings. AB - Preimplantation genetic diagnosis (PGD) was performed in two couples to avoid chromosomally unbalanced progeny in a family in which a brother and a sister carry an identical maternally inherited balanced translocation t(3;11)(q27.3;q24.3). Embryos were biopsied 3 days after fertilization and blastomeres were analysed by fluorescent in-situ hybridization (FISH). Embryos were classified as unbalanced or normal/balanced. In the first case, the male carrier and his wife underwent one IVF/PGD treatment cycle. In all, 18 embryos were analysed. Of those, 15 revealed an unbalanced karyotype. For one embryo, results were not conclusive, from one embryo results were contradictory and one embryo was classified as normal/balanced and subsequently transferred. A singleton pregnancy was achieved. The PGD analysis was confirmed at 16 weeks gestation by amniocentesis. At term, a healthy girl with a balanced karyotype was born. Pregnancy and delivery were without complications. In the second case, the female carrier and her husband underwent two IVF/PGD treatment cycles. During the first cycle, three embryos were analysed. One embryo revealed an unbalanced karyotype and two embryos were designated a normal/balanced karyotype and transferred but no pregnancy was achieved. During the second PGD cycle three embryos were analysed. Of those, none appeared suitable for transfer. The couple decided not to undergo further treatment. Our results indicate that for individuals carrying a reciprocal translocation PGD is a feasible approach to obtain embryos with a normal chromosome balance and to avoid both spontaneous and induced abortion. PMID- 10694264 TI - Quantitative analysis of mitochondrial Ca2+ uptake and release pathways in sympathetic neurons. Reconstruction of the recovery after depolarization-evoked [Ca2+]i elevations. AB - Rate equations for mitochondrial Ca2+ uptake and release and plasma membrane Ca2+ transport were determined from the measured fluxes in the preceding study and incorporated into a model of Ca2+ dynamics. It was asked if the measured fluxes are sufficient to account for the [Ca2+]i recovery kinetics after depolarization evoked [Ca2+]i elevations. Ca2+ transport across the plasma membrane was described by a parallel extrusion/leak system, while the rates of mitochondrial Ca2+ uptake and release were represented using equations like those describing Ca2+ transport by isolated mitochondria. Taken together, these rate descriptions account very well for the time course of recovery after [Ca2+]i elevations evoked by weak and strong depolarization and their differential sensitivity to FCCP, CGP 37157, and [Na+]i. The model also leads to three general conclusions about mitochondrial Ca2+ transport in intact cells: (1) mitochondria are expected to accumulate Ca2+ even in response to stimuli that raise [Ca2+]i only slightly above resting levels; (2) there are two qualitatively different stimulus regimes that parallel the buffering and non-buffering modes of Ca2+ transport by isolated mitochondria that have been described previously; (3) the impact of mitochondrial Ca2+ transport on intracellular calcium dynamics is strongly influenced by nonmitochondrial Ca2+ transport; in particular, the magnitude of the prolonged [Ca2+]i elevation that occurs during the plateau phase of recovery is related to the Ca2+ set-point described in studies of isolated mitochondria, but is a property of mitochondrial Ca2+ transport in a cellular context. Finally, the model resolves the paradoxical finding that stimulus-induced [Ca2+]i elevations as small as approximately 300 nM increase intramitochondrial total Ca2+ concentration, but the steady [Ca2+]i elevations evoked by such stimuli are not influenced by FCCP. PMID- 10694266 TI - The size of the CAG repeat in exon 1 of the androgen receptor gene shows no significant relationship to impaired spermatogenesis in an infertile Caucasoid sample of German origin. AB - The androgen receptor (AR) gene, located on the X-chromosome at Xq11-12, contains in exon 1 a polymorphic CAG repeat which codes for a polyglutamine tract. Contractions of the CAG repeat are said to be related to prostate cancer. In contrast, sizeable expansion of the CAG repeat can cause spinal and bulbar muscular atrophy (SBMA). In infertile patients of Chinese origin and in a Melbourne multinational population impaired sperm production has been postulated to be related to moderate expansions of the polyglutamine tract. In a study of a Swedish population of infertile patients these findings could not be corroborated. The aim of our investigation was to examine the correlation between the length of the CAG repeat and impaired sperm production in an infertile Caucasoid patient sample of German ethnic origin. We found no statistically significant relationship between the size of the CAG repeat or polyglutamine tract and idiopathic impaired sperm production in the population studied. The variability of the results by various investigators may be attributed to different ethnic origins and hence different genetic modifiers of the populations studied and/or to the high probability that these infertile males may represent a heterogeneous group with respect to the causes of defective spermatogenesis. PMID- 10694267 TI - Prostatic origin of a zinc binding high molecular weight protein complex in human seminal plasma. AB - The profile of the zinc ligand high molecular weight proteins was investigated in the seminal plasma of 55 normozoospermic subjects by size exclusion high performance liquid chromatography (HPLC). The proteins were recovered from Sephadex G-75 gel filtration of seminal plasma in three zinc-containing fractions which were then submitted to HPLC analysis. The results were, that in all the samples, the protein profiles showed two peaks with apparent molecular weight of approximately 660 and approximately 250 kDa. Dialysis experiments revealed that both approximately 660 and approximately 250 kDa proteins were able to uptake zinc against gradient indicating their zinc binding capacity. The HPLC analysis of the whole seminal plasma evidenced only the approximately 660 kDa protein complex as a single well quantifying peak, furthermore a positive correlation between its peak area and the seminal zinc values (P < 0.001) was observed. This suggested a prostatic origin of the approximately 660 kDa protein complex which was then confirmed by the seminal plasma HPLC analysis of a subject with agenesis of the Wolffian ducts. Finally the study demonstrated the presence of two zinc binding proteins, approximately 660 and approximately 250 kDa respectively, in human seminal plasma and the prostatic origin of the approximately 660 kDa. PMID- 10694268 TI - Expression of protamine-1 and -2 mRNA during human spermiogenesis. AB - During spermiogenesis, the histone-to-protamine replacement causes the compaction of the spermatid chromatin. The genes for protamines, PRM-1 and PRM-2, are transcribed in round and elongating spermatids. The transcripts are stored in a translationally-repressed state by the binding of protein repressors before being translated in elongating and elongated spermatids. RNA extracts from homogenized whole testis samples supply only average data, and cell-specific and stage specific expression cannot be addressed. Therefore, we used UV-laser-assisted cell-picking (UV-LACP) to select spermatids of defined differentiation steps. Subsequent reverse transcription-polymerase chain reaction (RT-PCR) with intron spanning primer pairs allowed the detection of DNA-free and pseudogene-free PRM-1 and PRM-2 cDNA. Additional in-situ hybridization with digoxygenin-labelled cRNA probes exhibited PRM-1 and PRM-2 mRNA from step 1/2 spermatids to step 4 spermatids, but not in elongated spermatids. RT-PCR revealed amplicons for PRM-1 and PRM-2 in all spermatids except step 3 round spermatids. Applying proteinase K digestion, PRM-1 and PRM-2 transcripts were also detected in step 3 spermatids indicating that protein repressors may bind to both PRM-1 and PRM-2 mRNA in step 3 round spermatids. These data demonstrate that the combination of UV-LACP and non-radioactive in-situ hybridization appear to be a suitable approach for the study of cell-specific and stage-specific gene expression during spermiogenesis. PMID- 10694269 TI - Fatty acid composition of spermatozoa and immature germ cells. AB - A great deal of attention has recently been given to the essential role of polyunsaturated fatty acids (PUFA) of sperm membranes. We studied the fatty acid composition of the immature germ cells (IGC) and of the sperm populations separated by Percoll gradient in the ejaculate of normozoospermic patients. Fatty acid pattern was analysed by combined gas chromatography-mass spectrometry on a capillary column. In IGC, differences were found compared with mature spermatozoa, with a higher percentage of saturated fatty acids and of essential fatty acids. On the contrary, the long-chain PUFA were significantly lower in IGC. The highest concentration of n3 PUFA docohexaenoic acid (DHA) was detected in the spermatozoa deriving from 70-100% Percoll layers and a direct linear correlation was found between the increase of DHA and increased percentage of Percoll gradient. An inverse relationship between the percentage of atypical sperm forms in each layer and the percentage of DHA was also observed. This study demonstrates that the human germ cell line can elongate and desaturate essential fatty acids and that the percentage of long-chain PUFA is correlated with the normal morphology of sperm cells. PMID- 10694270 TI - Sensitivity of mouse oocytes to nicotine-induced perturbations during oocyte meiotic maturation and aneuploidy in vivo and in vitro. AB - Oocyte meiosis is sensitive to endogenous and exogenous perturbations that upset the temporal sequence of biochemical reactions during oocyte maturation (OM) and predispose oocytes to aneuploidy. Nicotine is an alkaloid that has been reported to disrupt the rate of OM, reduce ovulation and fertilization rates, and increase diploidy. The objective of this study was to test the hypothesis that nicotine perturbs the rate of OM and induces aneuploidy in mouse oocytes in vivo and in vitro. Female mice were given 7.5 IU pregnant mare's serum and either 0, 5.0, 7.5, or 10 mg/kg nicotine in vivo at -3, 0, and +3 h relative to a 5 IU injection of HCG. Oocytes were also cultured in vitro in the presence of 0, 1.0, 5.0, or 10.0 mmol/l nicotine. In vivo, significant (P < 0.05) differences in the proportions of oocytes with premature centromere separation and premature anaphase were found at 10.0 mg/kg nicotine suggesting that the rate of OM was advanced. Also, at this dose the proportion of ovulated oocytes was reduced by approximately 50% relative to controls. In vitro, only non-significant differences were found among the parameters measured. Although nicotine reduced the ovulation rate and perturbed the rate of OM in vivo, these data show that the rate of aneuploidy was not significantly elevated. PMID- 10694271 TI - Involvement of progesterone in gonadotrophin-induced pituitary adenylate cyclase activating polypeptide gene expression in pre-ovulatory follicles of rat ovary. AB - The present study was designed to determine whether progesterone might have a role in gonadotrophin-induced pituitary adenylate cyclase-activating polypeptide (Pacap) gene expression in rat ovary. Northern blot analysis revealed that treatment of pregnant mare's serum gonadotrophin (PMSG)-primed immature rats with the progestin antagonist RU486 or an inhibitor of 3beta-hydroxysteroid dehydrogenase epostane, 1 h before HCG, resulted in a dose-dependent inhibition of the HCG-induced Pacap gene expression. In-situ hybridization demonstrated that the number of pre-ovulatory follicles expressing Pacap mRNA in their granulosa cells was greatly reduced in ovaries treated with RU486. Moreover, the suppressive effect of RU486 or epostane on the LH-induced Pacap gene expression in cultured pre-ovulatory follicles was reversed by co-treatment with the synthetic progestin R5020. We further cloned the 5'-flanking region of the rat Pacap gene and identified the presence of a consensus progesterone receptor element. When luciferase fusion genes containing Pacap gene promoter were transiently transfected into granulosa cells of pre-ovulatory follicles, luciferase activity was markedly stimulated by LH. Treatment with RU486 or epostane resulted in partial suppression of LH-stimulated PACAP promoter activity. Taken together, these results indicate that progesterone, acting through progesterone receptors, plays a role in gonadotrophin induction of Pacap gene expression in granulosa cells of pre-ovulatory follicles, and thereby may be involved in the process of ovulation. PMID- 10694272 TI - Effects of lipopolysaccharide and cytokines on production of RANTES by cultured human endometrial stromal cells. AB - RANTES (regulated upon activation, normal T cell expressed and secreted), which is a potent chemoattractant for eosinophils, lymphocytes, and monocytes, was recently detected in the human endometrium. The effects of modulators of endometrial function, including lipopolysaccharide (LPS), tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-4 and interferon-gamma (IFN-gamma), on the production of RANTES by endometrial stromal cells (ESC) were examined by an enzyme-linked immunosorbent assay and Northern blot analysis. The concentration of RANTES in the culture media of non-stimulated ESC was below the level of detection. The concentration of RANTES was increased by the addition of TNF alpha, IL-1beta and LPS. IFN-gamma synergistically enhanced the TNF-alpha- and LPS-induced RANTES expression, but had no effect on the IL-1beta-induced RANTES expression. The TNF-alpha-induced production of RANTES by ESC was inhibited by IL 4. The transcription of RANTES in ESC was also stimulated by TNF-alpha, IL-1beta and LPS in a dose-dependent manner. It is suggested that the LPS and cytokines secreted by the maternal decidual tissue and the developing embryo may regulate the production of RANTES by ESC. The modulation of RANTES concentration in the local environment may contribute to the pathophysiological processes of human reproduction by regulating the immunological reaction at the fetal-maternal interface. PMID- 10694273 TI - CD9 is expressed on human endometrial epithelial cells in association with integrins alpha(6), alpha(3) and beta(1). AB - Recently we reported that CD9 is involved in the invasion of a trophoblast-like choriocarcinoma cell line, BeWo, probably through the regulation of integrin functions. Integrins have also been reported to be expressed in the human endometrium and it has been suggested that they play important roles in blastocyst implantation. This study used immunohistochemistry to investigate the expression of CD9 in the endometrium during the menstrual cycle. CD9 was found to be intensely expressed on the cell surface of the glandular epithelium throughout the menstrual cycle without any apparent differences in staining intensity. In addition, Western blotting analysis of the affinity-purified proteins confirmed that CD9 was associated with integrins beta(1), alpha(3) and alpha(6) in the human endometrium. Therefore it can be concluded that CD9, in association with integrins alpha(6), alpha(3) and beta(1), is a constitutive molecule of the endometrial glandular epithelium. These results also suggest that CD9 may be an important regulator of these integrins in the human endometrium. PMID- 10694274 TI - Endometrial lysosomal enzyme activity in ovulatory dysfunctional uterine bleeding, IUCD users and post-partum women. AB - The aim of this study was to evaluate the role of lysosomal enzymes in excessively heavy menstruation by comparing women with menorrhagia due to dysfunctional bleeding or intrauterine contraceptive device (IUCD) use with those with normal menstrual periods or with amenorrhoea associated with breastfeeding. This was a prospective cohort investigation of the activity of four endometrial lysosomal enzymes in three contrasting groups: (i) women with ovulatory dysfunctional uterine bleeding and users of intrauterine contraceptive devices; (ii) breastfeeding post-partum women in whom there are long periods of amenorrhoea, particularly in the early months post-partum; and (iii) normal cycling women. It was found that the total activity of lysosomal enzymes, particularly acid phosphatase and N-acetyl-beta-D-glucosaminidase, was markedly elevated (P < 0.001) in IUCD-exposed endometrium, and endometrium from women with dysfunctional uterine bleeding when compared with endometrium from women with a history of entirely normal menstrual periods or that in post-partum breastfeeding women. The activity of alpha-L-fucosidase was moderately elevated in IUCD users (P < 0.05) and ovulatory dysfunctional uterine bleeding (P < 0.05), whereas alphaD-mannosidase activity was elevated in ovulatory dysfunctional uterine bleeding (P < 0.05), but decreased in IUCD users (P < 0.01). No significant differences were observed in the lysosomal enzyme activities of breastfeeding post-partum women and normal cycling women. These results show that total endometrial tissue activity of four lysosomal enzymes was substantially increased throughout the cycle in most circumstances in women with two different causes for increased menstrual bleeding. This suggests a contributory role to the increased bleeding. PMID- 10694275 TI - The progesterone receptor and ubiquitin are differentially regulated within the endometrial glands of the natural and stimulated cycle. AB - The initiation of human pregnancy requires precisely timed development of the endometrium to receive the implanting blastocyst. The ovarian steroid hormones are essential for development and maintenance of a hospitable uterine environment. The hormonal regimes employed in assisted reproduction procedures are known to alter the abundance of specific endometrial receptors for these steroids. Since, in the presence of ligand, the progesterone receptor (PR) is known to be modified by the small intracellular protein ubiquitin, we have investigated the localization of ubiquitin and PR within the endometrial glands of 28 fertile women during a monitored menstrual cycle and also during a stimulated cycle prior to oocyte donation. We have also observed the number of gland cells undergoing cell division as demonstrated by the presence of Ki67 immunostaining. We demonstrate that the percentage of ubiquitin-positive nuclei increases from day four post-ovulation to day 10 post-ovulation in the natural cycle, but that this increase is not seen during a stimulated cycle. The presence of PR within glandular epithelium and the proliferation of gland cells were only observed during the early secretory phase and did not appear to vary significantly between the two cycles. We conclude that ubiquitin may play an important role in endometrial development and that perturbation of ubiquitin may be related to the lower implantation rate seen in the stimulated cycle. PMID- 10694276 TI - Induction of an angiogenic phenotype in endometriotic stromal cell cultures by interleukin-1beta. AB - Activated peritoneal macrophages are associated with endometriosis and may play a central role in its aetiology by releasing interleukin-1beta (IL-1beta) in response to refluxed endometrium. Pari passu with the establishment of endometriotic implants is the development of a vascular supply. In this study we investigated the angiogenic properties of two endometrial proteins, vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6), and assessed their production in response to IL-1beta stimulation in human stromal cells isolated from normal endometrium (NE) and endometriotic lesions (EI). Proliferation of bovine brain capillary endothelial cells (BBCE) with a [(3)H]-thymidine incorporation assay was observed when VEGF (2.1 +/- 0.2-fold; P < 0.05) or VEGF and IL-6 (1.8 +/- 0.1-fold; P < 0.05) were added in vitro, relative to saline treated control cultures. Northern blot analysis showed induction of VEGF mRNA (2.6-fold; P < 0.05) and IL-6 mRNA (6.3-fold; P < 0.05) transcripts in EI cells, but not NE cells, exposed to IL-1beta. A similar induction was seen with VEGF and IL-6 protein secretion in the responsive EI cells. Reverse transcription polymerase chain reaction (RT-PCR) for the IL-1 receptor type I (IL-1 RI) indicated that the differential effects of IL-1beta on NE and EI cells was associated with 2.4 +/- 0.1-fold more receptor mRNA in EI versus NE cells. We propose that the ability of IL-1beta to activate an angiogenic phenotype in EI stromal cells but not in NE cells, is mediated by the IL-1 RI. PMID- 10694277 TI - VEGF, its receptors and the tie receptors in recurrent miscarriage. AB - The aetiology of recurrent miscarriage (at least three consecutive miscarriages) usually remains unsolved. The vascular endothelial growth factor (VEGF) family of proteins, together with their receptors and the Tie (tyrosine kinase with immunoglobulin and epidermal growth factor homology domains) receptors, are crucial for embryonic development. Therefore, we used immunohistochemistry to analyse the expression of VEGF, the VEGF receptors (VEGFR)-1, -2, and -3, and the Tie-1 and Tie-2 receptors in placental and decidual tissue of women with a history of recurrent miscarriage and missed abortion (MA; n = 12) or blighted ovum (BO; n = 6), and from normal early terminated pregnancies (n = 12). Compared with controls, the MA and BO groups showed: (i) diminished placental trophoblastic VEGF immunoreactivity; (ii) weaker VEGFR-1 and -2 immunoreactivity in decidual vascular endothelium; (iii) reduced placental trophoblastic Tie-1 receptor immunoreactivity; and (iv) reduced decidual vascular endothelial Tie-1 and -2 receptor immunoreactivity. The absence of VEGFR-3 immunoreactivity in decidual vascular endothelium was also noted in all study groups. Interestingly, placental villi from the BO group presented blood vessel-like structures negative for von Willebrand factor, but positive for VEGF, VEGFR-1, -2, -3, Tie-1 and Tie 2 receptor. We conclude that the expression of these antigens may be altered in recurrent miscarriages. PMID- 10694278 TI - Expression and localization of inducible nitric oxide synthase in human non pregnant and early pregnant endometrium. AB - The aim of the present study was to investigate the expression and distribution patterns of inducible nitric oxide synthase (iNOS) in human non-pregnant and early pregnant endometrium using Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. Northern blot analysis revealed the expression of iNOS mRNA in human decidua and chorionic villi in the first trimester but not in the endometrium at any stage of the menstrual cycle. Nested RT-PCR, however, detected iNOS mRNA in human endometrium at all stages of the menstrual cycle. Immunohistochemical staining of the secretory endometrium using an anti-human iNOS polyclonal antibody revealed labelling specifically concentrated in glandular epithelial cells. Staining was absent in stromal cells. However, iNOS staining was positive in decidualized stromal cells in tissues obtained in the first trimester of pregnancy. Furthermore, extensive staining was observed in both syncytiotrophoblastic and cytotrophoblastic cells. The finding of a large amount of iNOS mRNA at the feto maternal interface throughout the first trimester of pregnancy suggests that iNOS may play an important role in the maintenance of pregnancy. PMID- 10694279 TI - Assessment of medical journal quality. PMID- 10694280 TI - Osteonecrosis, corticosteroid use and Crohn's disease: evidence-based medicine versus civil law. PMID- 10694281 TI - Use of community resources before inflammatory bowel disease surgery is associated with postsurgical quality of life. AB - BACKGROUND: Research in chronic illness shows that community resources can have a lasting influence on the course of the illness; however, little research has been done to evaluate the community agencies that specifically address the needs of inflammatory bowel disease (IBD) patients. OBJECTIVES: To survey awareness of community agency resources among patients who have surgery for IBD, and to analyze the association between using these resources and qualitative postsurgical outcomes. SUBJECTS AND METHODS: Ninety-two subjects who had surgery over a 12-month period completed, in full, the Inflammatory Bowel Disease Questionnaire (IBDQ), and a self-report instrument used to probe awareness and use of local community resources. Community resources were divided into two groups: those involving primarily social and educational participation ('social/ educational') and those involving some individualized attention, usually from a professional or trained lay facilitator ('professional/individual'). The contribution of presurgical participation in each type of resource to postsurgical quality of life was tested using ANOVA, with IBDQ score as the dependent variable. The ANOVA was repeated with postsurgical disease activity as a covariable. IBDQ subscale scores were compared between groups that were found to differ in the ANOVA. RESULTS: Almost all subjects were aware of at least one available resource. Participation in resources before surgery was variable, but 50% of the sample participated in at least one social/educational resource, and 46.9% participated in at least one professional/individual support. For the 92 subjects who completed both the IBDQ and the survey of resources, ANOVA revealed a main effect of professional/individual resource use on postsurgical quality of life but no main effect of social/educational resources and no interaction. DISCUSSION: The association between presurgical participation in professional or individualized community resources and better subjective outcome of IBD surgery may be explained by a positive contribution of participation to coping with surgery for IBD. The data do not support the alternative explanation that subjects with less severe disease (and thus better outcome) have greater ability to participate, although further research is required. PMID- 10694282 TI - Applications of recombinant DNA technology in gastrointestinal medicine and hepatology: basic paradigms of molecular cell biology. Part A: eukaryotic gene structure and DNA replication. AB - Progress in the basic sciences of cell and molecular biology has provided an exciting dimension that has translated into clinically relevant information in every medical subspecialty. Importantly, the application of recombinant DNA technology has played a major role in unravelling the intricacies related to the molecular pathophysiology of disease. This series of review articles constitutes a framework for the integration of the database of new information into the core knowledge base of concepts related to the pathogenesis of gastrointestinal disorders and liver disease. The goal of this series of three articles is to review the basic principles of eukaryotic gene expression. The first article examines the role of DNA in directing the flow of genetic information in eukaryotic cells. PMID- 10694283 TI - Obscure gastrointestinal bleeding: an approach to management. AB - Obscure gastrointestinal bleeding provides an uncommon but frustrating and resource-intensive challenge for clinicians. Such patients hemorrhage recurrently from sites within the gastrointestinal tract that are not detected by routine endoscopy or radiography, and require a special diagnostic approach to localize or exclude less common bleeding sources such as small bowel angioectasia or neoplasia. The differential diagnosis of obscure gastrointestinal hemorrhage is discussed, and the performance of available endoscopic, radiological and surgical diagnostic tools including enteroscopy are examined critically. A stepwise management algorithm that progresses from the history and physical examination to surgical exploration is offered to facilitate early and efficient diagnosis. PMID- 10694284 TI - HFE-associated hereditary hemochromatosis. AB - Hereditary hemochromatosis is a common inherited disorder of the iron metabolism. Screening studies indicate that it has a prevalence of one in 200 to 400, depending on the population studied, and a carrier rate of about one in seven to one in 10. Feder et al identified the hereditary hemochromatosis gene (HFE) in 1996 and two candidate mutations; the C282Y mutation has been shown to be responsible for the majority of the hereditary hemochromatosis cases worldwide. The gene discovery has led to rapid advances in the field of iron metabolism. Although the basic defect is still not fully understood, much is known about the sequence of events leading to iron overload. Hereditary hemochromatosis is a major candidate for population screening and meets the screening criteria of the World Health Organization, and Wilson and Jungner. It is one of the most prevalent genetic diseases in white populations, and, importantly, early diagnosis and simple effective treatment allow normal life expectancy. The discovery of the HFE gene and the frequency of the single C282Y mutation as a cause of most cases of hereditary hemochromatosis allow the possibility of widespread genetic testing. However, the logistics, and the psychological and social consequence of this, coupled with incomplete expression of the genotype, necessitate further studies before population screening can be justified. PMID- 10694285 TI - Benign pancreatic duct strictures: medical and endoscopic therapy. AB - Pancreatic duct strictures usually reflect underlying pancreatic disease and are likely caused by one or more of the following: acute or chronic pancreatitis, benign or malignant pancreatic neoplasm, pseudocyst and trauma. The characteristics of pancreatic strictures are identified, and medical and endoscopic therapy options are reviewed. PMID- 10694286 TI - Update on gastroesophageal reflux and respiratory disease in children. AB - Pediatric respiratory diseases have been linked to gastroesophageal reflux disease (GERD), but evidence regarding the association and its potential mechanisms continues to accumulate, and important aspects remain to be determined. Evidence for the association in two common pediatric respiratory disorders - infantile apnea and asthma in older children - and difficult clinical issues associated with the diagnosis and treatment of these two disorders are reviewed. The provocative embryological and physiological connections between the upper gastrointestinal tract and the respiratory tract, and recent understanding of the compensatory anatomy and physiology that protect the normal individual from respiratory manifestations of GERD are also explored. Dysfunctions of these protections likely underlie the pathophysiology of these disorders. PMID- 10694287 TI - Prevalence rates and an evaluation of reported risk factors for osteonecrosis (avascular necrosis) in Crohn's disease. AB - Avascular necrosis (osteonecrosis) occurs in Crohn's disease, but the rate of this particular complication is not known. Over 20 years, 877 patients with Crohn's disease, 492 women (56.1%) and 385 men (43.9%), were evaluated with patient follow-up data available for a mean of 7.8 years. In this group, four men were seen with osteonecrosis. No woman was affected. All patients had typical radiological, magnetic resonance imaging or pathological changes of osteonecrosis involving the femoral heads, while two also had superimposed avascular necrosis involving the humeral heads. Patient ages ranged from 19 to 36 years at the time of diagnosis of their Crohn's disease, and all were white. In one patient, disease was confined to the colon, while three patients had disease involving the terminal ileum and colon. Disease behaviour in two patients was classified as penetrating because of concomitant ischiorectal abscesses, while one patient developed a metastatic colon carcinoma. Ankylosing spondylitis was present in two patients, but no other extraintestinal manifestations developed. Two patients received corticosteroids as well as parenteral nutrition during the course of their disease. Two patients did not receive corticosteroids or parenteral nutrition. Of 877 patients with Crohn's disease, 484 (55. 1%) received corticosteroids during the course of the disease, 196 (22.4%) received at least one course of parenteral nutrition, and 125 (14.3%) received both corticosteroids and parenteral nutrition. A total of 311 patients (35.5%) had at least one small intestinal resection. The overall rate of avascular necrosis in Crohn's disease was less than 0.5% but for men with Crohn's disease was about 1%. In this series, risk of osteonecrosis could not be attributed to corticosteroid use, parenteral nutrition or both forms of therapy administered together. Small intestinal resection with loss of small intestinal absorptive area was not a risk factor for the development of osteonecrosis. Avascular necrosis (or osteonecrosis) is a very rare extraintestinal osseous complication that may occur in Crohn's disease, independent of previously reported risk factors, including corticosteroids or parenteral nutrition with lipid emulsions. PMID- 10694288 TI - Taking action: implementing your written investment plan. PMID- 10694289 TI - Founding of a new european dermatology organisation PMID- 10694290 TI - The future of dermatology in Europe: position paper. PMID- 10694291 TI - Coagulation factor XIII, endothelial damage and systemic sclerosis. AB - In the blood coagulation process, Factor XIII (F XIII) is responsible for the stabilization of the fibrin clot. The hypothesized ability of this blood coagulation factor to affect collagen synthesis and degradation led to its use in the treatment of scleroderma. However, the complex mechanism of action of F XIII remains unclear. The aim of our study was to assess possible effects of F XIII on endothelial damage, regarded as an early pathogenic event in systemic sclerosis (SSc). Thus, we measured plasma levels of von Willebrand factor antigen (vWF:Ag), a marker of endothelial cell injury, in 22 patients with SSc, 9 of whom were treated with F XIII and 13 who do not receive the drug. The plasma concentration of F XIII has also been analyzed. Interestingly, the vWF:Ag plasma levels within the group of SSc patients treated with F XIII were significantly lower than those of untreated SSc individuals (p < 0. 02). On the other hand, the highest mean value of vWF:Ag was found in a subset of untreated subjects having SSc with severe lung involvement, supporting a strict relationship between elevated levels of vWF:Ag and severity of the disease. In contrast, plasma concentration of F XIII resulted normal in all but 3 SSc patients, ruling out a deficiency of this blood coagulation factor as promoting the occurrence of SSc. These preliminary findings seem to support the hypothesis that F XIII may play an improving role on endothelial damage, other than the initially suggested action on collagen metabolism, in SSc. PMID- 10694292 TI - An experience for ELISA for desmoglein 1, suggesting a possible diagnostic help to determine the initial therapy for pemphigus foliaceus. AB - Pemphigus foliaceus (PF) is an autoimmune skin disease in which loss of adhesion between keratinocytes (i.e., acantholysis) in the granular cell layers appears to be mediated by the binding of autoantibodies against desmoglein (Dsg) 1. Although it has been suggested that the activity of the disease is rather correlated with the titer of the antibody, there are still no precise studies on the relationships between the disease activity and the titers of autoantibody against Dsg1 by using ELISA. In this study, we performed ELISA using recombinant Dsg1 (rDsg1) as antigen on 8 patients with pemphigus foliaceus in order to examine the reliability of this test for monitoring disease severity. The resultant index values of ELISA, which were defined by the formula: (OD of tested serum - OD of negative control)/(OD of positive control - OD of negative control) X 100, ranged from 16 to 264 compared with 6.41 of cutoff index value for normal sera. A patient with an index value of 213 had severe widespread erosions with erythema and required pulse therapy of methylpredonisolone (1,000 mg/day for 3 days). A patient with an index value of 111 had a moderate severity and was successfully treated with an initial dose of 30 mg/day (0.64 mg/kg) of predonisolone. The mildest patient with an index value of 16 was controlled with only topical application of 0.05% betamethasone ointment twice a day. These results suggest that ELISA index values for rDsg1 will provide a promising tool for monitoring the disease severity and for determining the initial therapy for pemphigus foliaceus. PMID- 10694293 TI - Clinicopathological correlation of pigmented skin lesions using dermoscopy. AB - Dermoscopy (dermatoscopy, epiluminescence microscopy) is an additional measure for making the diagnosis of pigmented skin lesions more accurate. It enables the clinician to visualize features not discernible by the naked eye. By applying enhanced digital dermoscopy and a standardized gross pathology protocol to pigmented skin lesions, a precise clinicopathological correlation of relevant dermoscopic features can be made. Histological specimens of four pigmented skin lesions (melanoma in situ, Clark's nevus, Reed's nevus, seborrheic keratosis) were processed using a standardized gross pathology protocol and viewed along with the clinical photographs and digital dermoscopic images that were magnified and enhanced to better visualize the corresponding dermoscopic structures. Furthermore, measurements of dermoscopic structures using digital equipment were correlated with histometric findings. Our understanding of dermoscopic features, especially the broadened pigment network - a specific dermoscopic criterion for melanoma - was refined by this detailed case-by-case correlation. In addition, some not yet fully characterized dermoscopic features, such as black lamella, radial streaks, and exophytic papillary structures, were described in detail dermoscopically and histopathologically. Moreover, measurements of these dermoscopic features and the underlying histological structures were found to be similar. Linking dermoscopy more closely with cutaneous pathology may help refine the definitions and diagnostic criteria of pigmented skin lesions for dermatologists as well as dermatopathologists. PMID- 10694294 TI - High dose intravenous immunoglobulin (IVIG) in dermatomyositis: clinical responses and effect on sIL-2R levels. AB - An open study was conducted to identify and investigate dermatomyositis patients who benefit from IVIG treatment, based on dermatological criteria, myositis related symptoms and immune/inflammatory parameters. 19 patients (16 females and three males, ages 31-84) suffered from dermatomyositis, and 4/19 patients had paraneoplastic dermatomyositis. We monitored the disease activity by documenting the clinical symptoms, recording muscle-related parameters (electromyography, serum creatine kinase, histopathology), and by determining circulating autoantibodies and serum levels of IL-6, sIL-2R, sTNF-a-R, sICAM-1, and sCD8. 7/19 patients responded to IVIG. They had severe skin but only moderate muscle involvement, no autoantibodies, and no malignancy. IVIG-nonresponders had severe skin and muscle disease, concomitant with autoantibodies and/or malignancy. sIL 2R levels were initially elevated in all patients but reverted to normal in IVIG responders only. Creatine kinase-levels and other parameters did not correlate with disease activity and/or treatment response. IVIG is effective in selected dermatomyositis patients. sIL-2R serum levels appear to be useful predictors of IVIG-induced treatment response and disease activity. PMID- 10694295 TI - Recent physiopathological insights in cutaneous lymphocytic infiltrates AB - As there were typing errors in the summary of the article by Herve Bachelez in the EJD vol. 9, n 7, a corrected version is published below. The spectrum of benign cutaneous lymphocytic infiltrates (CLI) includes a variety of entities which are classified on the basis of clinicopathological findings, and of the phenotype of the predominant lymphocytic subset among the skin infiltrate. Major concern has been recently given to the clonality status of CLI by using highly sensitive assays based on the PCR amplification of TCR/Ig loci. These studies allowed the characterisation of clonal benign cutaneous lymphocytic expansions. Other studies have shown evidence of oligoclonal patterns in HIV-associated CD8 cutaneous pseudolymphomas, and functional studies of the skin infiltrate further showed that an antigen-driven mechanism was involved in the pathogenesis of this latter entity. Finally, knowledge in the field of CLI has been improved by the identification of antigens associated with skin-homing properties such as the so called cutaneous lymphocyte-associated antigen (CLA) which is expressed at the surface of most memory T cells infiltrating the dermis in inflammatory conditions. PMID- 10694296 TI - Acrogeria of the Gottron type in a mother and son. AB - We report a familial case of acrogeria in a mother and son, with characteristic cutaneous involvement and no clinical signs of vascular Ehlers-Danlos syndrome (former EDS type IV) in spite of some tendency to bruising. The biochemical and molecular studies did not disclose any abnormality of collagen type III, which favours the diagnosis of acrogeria. It appears that recognition of acrogeria as an entity is of clinical significance since these cases are not associated with systemic involvement, and specifically with rupture of vessels and internal organs, occasionnally occurring in EDS. PMID- 10694297 TI - Correlation between bacterial population and axillary and plantar bromidrosis: study of 30 patients. AB - Although studies on the chemistry of odors are expanding to identify the chemical structures of odorous substances, there are no universal standards as yet to measure odor and intensity of bromidrosis. Clinical evaluation can be made on a subjective scoring from 0 to 3 prior to prescription of an antiseptic soap. In order to appreciate the correlation between the intensity of bromidrosis (BI) and bacterial activity, a study was carried out with both clinical and bacterial assessment in thirty patients with axillary or plantar BI. Odor intensity was evaluated by two physicians using a score from 0 to 3 (i.e. absent, minor, moderate, major), meanwhile bacterial composition and density were assessed before and after 10 days of hygiene using an antiseptic detergent (trichlocarbanilide) provided on the first visit. Baseline count of diphtheroids/cm2 was 35.104 and baseline micrococci average was 32.104/cm2. At the end of the study, the reduction of odor intensity was observed in 20 patients (67%) without any change in sweat production. The clinical improvement correlated with a reduction of both micrococci (70%) or diphtheroids (73%) as compared with initial data. In patients presenting persistant bromidrosis, the bacterial count/cm2 did not significantly decrease and remained above 104 diphtheroids/cm2. Thus, this study suggests that body odor may be at least indirectly correlated to microbia counts with a bacteria threshold of BI ranging around and above 104. PMID- 10694298 TI - Occurrence of Hodgkin's disease and cutaneous B-cell lymphoma in the same patient: a report of two cases. AB - The occurrence of Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) in the same patient is well known. The most frequent observation has been the development of large B cell lymphoma in patients affected with the nodular form of lymphocytic predominant HD. A less common situation is the development of NHL among patients successfully treated for HD. In such patients the second lymphoma has been thought to be related to the previous therapy or the immunodeficiency state that can accompany HD. Histologically, these NHL lymphomas often are intermediate to high grade and frequently extranodal. We report two patients successfully treated for HD who also developed NHL of the skin. Both patients presented with strikingly similar findings regarding to sex, age and subtype of HD. Clinical, histopathological and immunophenotypical findings were consistent with cutaneous low-grade B cell lymphoma of the marginal zone type. Both cases remain in complete remission of HD after standard therapy. In both patients the cutaneous lymphoma followed an indolent clinical course after a long follow-up period. This observation expands the spectrum of alterations possibly related to HD. PMID- 10694299 TI - Pulmonary nocardiosis with cutaneous involvement mimicking a metastasizing lung carcinoma in a patient with chronic myelogenous leukaemia. AB - We report a unique case of a man suffering from chronic myelogenous leukaemia who presented with clinical symptoms, X-ray, and bronchoscopical findings consistant with a bronchopulmonary space-occupying process which was suspected to be a central lung carcinoma as a secondary de novo malignancy. In addition, the patient developed several subcutaneous nodular livid red lesions on the left forearm which were considered to be cutaneous metastases of the presumptive lung malignancy. Treatment was started with percutaneous radiation of the mediastinum over a period of ten days with a total dose of 25 Gray. The patient died from circulatory and respiratory failure. Only post mortem pathological examination was indicative of a nocardiosis of the lungs with haematological spread to eosophagus, pleura, and subcutaneous skin of the left forearm. Unfortunately, diagnosis of nocardiosis could not finally proven by culture or molecular biological methods. A lung carcinoma or an infiltrate of residual or relapsing chronic myelogenous leukemia in the lung could be definitely ruled out. PMID- 10694301 TI - Raynaud's phenomenon possibly induced by a compund drug of tegafur and uracil. AB - We describe a 50-year-old woman who noted acral hyperpigmentation, sclerodactyly and Raynaud's phenomenon with 1:160 of antinuclear antibody titer after treatment with a compound drug with tegafur and uracil. Histological findings of the finger and palm included hyperkeratosis, vacuolar degeneration of basal cells, thickened collagen fibers in the dermis, and dilatation of capillary vessels, perivascular mononuclear cell infiltration and melanophages in the upper dermis. IgG, IgA, IgM, C3 and C1q were not deposited in the skin by direct immunofluorescence study. After cessation of the drug, Raynaud's phenomenon and hyperpigmentation disappeared within 1 month and 4 months, respectively, and antinuclear antibody turned negative within 4 months. These observations suggest that tegafur may have caused not only hyperpigmentation in the palms and soles, but also sclerodactyly and Raynaud's phenomenon in the present case. PMID- 10694300 TI - Tumoral calcinosis: a case report with an electron microscopic study. AB - A 68-year-old woman developed large subcutaneous masses on her abdomen and thighs after a bruise sustained in a traffic accident. She had severe pain when sitting up straight. Histological examination revealed calcified tissues in the entire dermis of the injured areas. On electron microscopy, crystalline materials were observed in the dermis, which seemed to be formed by the deposition of hydroxyapatite on unusual proteoglycan. In a vessel wall, a thick, layered basement membrane was observed. This suggests that vascular injury and subsequent hypoxia play a role in the process of calcinosis. We performed a partial resection with good results in alleviating the patient's pain. PMID- 10694302 TI - Treatment of pruritus of reactive perforating collagenosis using transcutaneous electrical nerve stimulation. AB - Reactive perforating collagenosis is a form of perforating dermatosis due to transepithelial elimination of collagen and characterized by itchy papulonodular eruptions frequently seen in patients with diabetes mellitus and end stage renal failure. Pruritus is often severe and treatment is difficult. Two adult Chinese diabetic patients with acquired reactive perforating collagenosis unresponsive to topical therapies and oral antihistamines, were treated with transcutaneous electrical nerve stimulation. There was a significant reduction of itch followed by gradual resolution of the skin lesions. PMID- 10694303 TI - Understanding the trends in melanoma incidence and mortality: where do we stand? AB - In previous decades melanoma incidence rates have risen spectacularly in white populations worldwide and a parallel - although more moderate - increase has been observed for melanoma mortality. More recently several reports have been made of a stabilization or decrease of mortality rates in the younger birth cohorts, resulting in a stabilization in the overall mortality in the 1980s for some populations. This article reviews past and current trends in melanoma epidemiology. It further handles the possible explanations for these trends and the currently available indications for or against these trends being brought about by real or artefactual influences. Time will undoubtedly reveal more about the truth. PMID- 10694304 TI - Nerve growth factor and keratinocytes: a role in psoriasis. AB - Nerve growth factor (NGF) is synthesized and released by human keratinocytes. NGF acts as a neurotrophic molecule at the skin level, as it stimulates the sprouting of nerve fibers and regulates the synthesis and expression of neuropeptides. NGF can thus take part in neurogenic inflammation which in turn is involved in the pathogenesis of several inflammatory dermatoses. Human keratinocytes also synthesize and express the low (p75)-and the high-affinity (trk) NGF-receptor (NGF-R). NGF stimulates keratinocyte proliferation which is blocked by the natural alcaloid K252, a specific inhibitor of trk phosphorylation. K252 inhibits keratinocyte proliferation and induces keratinocyte apoptosis, in the absence of exogenous NGF, indicating the existence of an autocrine loop where NGF and trk act as key players. Finally, NGF protein levels are increased in psoriatic as compared to non-lesional and normal skin, and psoriatic keratinocytes express higher amounts of NGF than normal keratinocytes. This review will discuss the above findings in view of a possible involvement of NGF in the pathomechanisms associated with the development of the psoriatic lesion. PMID- 10694305 TI - Pili torti and sensorineural hearing loss. A follow-up of Bjornstad's original patients and a review of the literature. AB - In 1965, Bjornstad described 8 patients with pili torti, of whom five also suffered from hearing loss. The combination of these two findings was later coined Bjornstad's syndrome. Typically, these patients develop hair loss in the first two years of life, while the hearing deficit may become evident in the first three to four years of life. However, considerable differences regarding age of onset and clinical severity have been reported, a pronounced hair shaft abnormality is often associated with severe hearing deficits. In a recent study of a Mexican family with pili torti and deafness, the inheritance was determined to be autosomal recessive, and mapped to the gene locus 2q34-q36. Hypogonadism and mental retardation are associated findings that have been described in patients with Bjornstad syndrome. In this re-investigation of Bjornstad's original patients, two additional patients with pili torti and hearing loss are described, and a review of the published cases of Bjornstad's syndrome is given, as well as a short overview of syndromes and conditions with twisted hairs. PMID- 10694306 TI - Atrichia, ichthyosis, follicular hyperkeratosis, chronic candidiasis, keratitis, seizures, mental retardation and inguinal hernia: a severe manifestation of IFAP syndrome? AB - A boy with congenital atrichia, ichthyosis follicular, keratitis, cutaneous infections and a huge inguinal hernia, but without deafness is reported. We believe it represents a new case of a rare X-linked recessive syndrome known as ichthyosis follicularis, alopecia, photophobia syndrome (IFAP). The differential diagnosis from keratitis ichthyosis deafness is discussed. The cutaneous infections seen in our case suggest the possibility of considering a genetic link between these syndromes. PMID- 10694307 TI - Temporary hair loss using the long-pulsed alexandrite laser at 20 milliseconds. AB - Facial hypertrichosis presents an enormous psychological burden for women. Temporary hair removal (waxing, plucking, etc.) and electrolysis are prolonged and unsatisfactory methods of treatment. For a few years several laser systems with varying wavelengths, pulse durations and energy fluences have been used successfully in laser epilation. In the retrospective study on hand, we report on results of 30 female patients with hypertrichosis in the facial area treated with the long pulse alexandrite laser at 20 msec (Cynosure PhotoGenica LPIR/Apogee; 755 nm; 20 msec; up to 30 J/cm2; 10 or 12.5 mm beam diameter) over an 18 month treatment period. After an average of 8 treatments, an average clearance rate of 75% could be achieved. Fair hair (white/blond/red) only showed a clearance rate of 10%. Hypo- and hyperpigmentation did not appear. The most frequent adverse effects were the occasional appearance of scattered crusting (17%), which healed without consequences, and folliculitis (13%). The average post-treatment observation time lasted 3.25 months. The long-pulsed alexandrite laser at a pulse duration of 20 msec is an effective and safe method of treatment of hypertrichosis in the facial region of women. Black hair responds considerably better to the laser treatment than fair hair. A longer post-treatment observation time is necessary, though, in order to provide evidence for the permanence of the success of the method. PMID- 10694308 TI - Comparative study between terbinafine 1% emulsion-gel versus ketoconazole 2% cream in tinea cruris and tinea corporis. AB - An open, prospective, comparative, randomised and parallel-group study of 65 patients was conducted to evaluate the efficacy and safety of topical 1 % emulsion-gel of terbinafine versus 2% ketoconazole cream in the treatment of tinea corporis and tinea cruris. Treatment for terbinafine emulsion-gel was applied once daily for 1 week, whereas ketoconazole cream was applied once daily for 2 weeks; patients were followed for 2 weeks. Thirty-three patients in the terbinafine group and 32 in the ketoconazole group were evaluated for efficacy and safety. At the end of the study, rates of mycological cure were 94% for terbinafine emulsion-gel and 69% for ketoconazole cream (p = 0.027). A clinical and mycological overall evaluation was obtained for 72% of patients receiving terbinafine emulsion gel and 31% of patients receiving ketoconazole cream (p = 0.002). A total of four patients (1 in the terbinafine group and 3 in the ketoconazole group) had contact dermatitis-like side effects. We conclude that a 1-week course of terbinafine 1% emulsion-gel is significantly more effective than ketoconazole 2% cream in the treatment of tinea corporis and tinea cruris as regards clinical and mycological cure and treatment safety. PMID- 10694309 TI - Reproducibility of a dermoscopic method (7FFM) for the diagnosis of malignant melanoma. AB - Dermoscopy improves sensitivity in the diagnosis of melanoma. We have developed a new diagnostic dermoscopic method which evaluates only seven dermoscopic features or criteria. We called the method Seven features for melanoma (7FFM). To present and to evaluate the reproducibility of our dermoscopic diagnostic method (7FFM) we held eight dermoscopic courses from 10/4/96 to 04/03/98 in various Italian cities. In fact the reliability of a diagnostic test or method mainly depends on the level of agreement in the interpretation of results among different observers. Only methods with good agreement can be used in clinical practice for the diagnosis of melanoma. Many dermatologists (207) attended one of the eight dermoscopic courses: each course was one-day in length and at the end of the course the participants evaluated a set of 25 dermoscopic slides using our dermoscopic method. Percentages of concordance and K values for a kappa statistical analysis to evaluate inter-rater reliability have been calculated. The method showed a mean percentage of concordance of 85.7%, median 88%, a mean K value of 0.699, median 0.684. These data point to a good agreement level. Our method shows good reproducibility after a short training program. PMID- 10694310 TI - Different interaction of mast cells with human endothelial cells and fibroblasts. AB - In a number of chronic inflammatory conditions resulting in fibrosis, perivascular mononuclear cell infiltration including mast cells (MC) has been shown before the onset of vascular injury and interstitial fibrosis. These observations suggest a role for MC in inducing endothelial cell (EC) injury and fibroblast (FB) proliferation and collagen synthesis. In view of these observations, the interactions of MC with EC and FB were studied. MC adhesion to EC and FB showed time-dependent increase reaching a plateau at 1 and 3 hrs, respectively. With added MC, the proliferation of EC showed a dose-dependent decrease, but that of FB, a dose-dependent increase. MC, MC surpernatant and sonicated MC induced dose-dependent cytotoxic activity to EC, whose cytotoxicy was inhibited by trypsin inhibitor. FB cocultured with MC showed 9.95 times collagen synthesis and 11.0 times protein synthesis compared with FB without MC. These results showed that 1) MC attached to EC inhibited the proliferation by cytotoxic activity to EC, which was due to a kind of proteolytic enzyme involving trypsin, 2) MC had proliferative and collagen synthetic activity to FB. These results suggest the possibility that MC have a role in a number of chronic inflammatory diseases resulting in vascular injury and interstitial fibrosis. PMID- 10694311 TI - The effects of hair loss in European men: a survey in four countries. AB - Despite the high prevalence and the accepted psychological aspects of male pattern hair loss, few have characterized the effects of hair loss in representative samples of men in different countries. A representative sample of households in 4 European countries (France, Germany, Italy and the United Kingdom) was contacted by an interviewer and resident males 18-40 years of age (n = 1,717) completed a questionnaire designed specifically to evaluate attitudes to hair loss. The questionnaire was comprised of 78 questions translated and pilot tested using standard methodology into each local language. Questionnaires queried about self-rated hair loss, satisfaction with hair appearance, noticeability of hair loss to others, and bother, concerns and perceptions about hair loss, as well as general physical health (the SF-12 questionnaire) plus three additional questions about mental health. The self-reported degree of hair loss in men was statistically significantly associated (p < 0.001) with all hair loss specific effects measured, except "limiting job opportunities". The impact of hair loss was generally consistent in the four countries surveyed, although less pronounced in the United Kingdom. Age was significantly correlated with hair loss (rs = 0.34, p < 0.001). Men with greater hair loss were more bothered, more concerned about looking older due to their hair loss, and less satisfied with their hair appearance. Male pattern hair loss has significant negative effects on hair-loss specific measures in men 18 to 40 years of age in France, Italy, Germany and the UK. The degree that hair loss is perceived as noticeable to others appears to be a significant contributor to these negative effects. PMID- 10694312 TI - Epidemiology of occupational contact dermatitis in a North Italian population. AB - Occupational contact dermatitis (OCD) is a very important skin disease both for its high frequency and for its social and economic implications. The aim of our work is to evaluate the epidemiology of occupational contact dermatitis in a north-Italian population and the possibility of a correct etiological diagnosis using the patch test standard series of GIRDCA (Italian Group of Resarch on Contact Dermatitis). We patch tested 1,565 out-patients affected by dermatitis with standard series GIRDCA and with other specific professional haptens. The manifestations were suspected of being of occupational origin by a dermatologist on the basis of clinical and anamnestic data. Of all the recorded professions we have considered only the more numerically significant: food industry, building industry, textile industry, employees, cleaners, hospital personnel, hairdressers, housewives, mechanics and metallurgists. Sixty-nine percent of contact dermatitis was found in women, the hairdressers had the greatest number of patients in the younger group (68.7% in the 11-20 years age group) and the textile industry workers in older group (100% in the 41-50 years age group). A positive allergological anamnesis emerged in 32.3% of allergic contact dermatitis. Irritant contact dermatitis (10.6%) was more frequent than allergic contact dermatitis (8.4%). The hands are the most common localization (94. 4%). The allergen with the highest frequency of positive reactions is p phenylenediamine (25.3%). We discuss the frequency of positives to various groups of allergens in each profession and the principal means of contact. Because of the frequency of this type of occupational skin disease, we stress the importance of prevention. The standard series GIRDCA was found to be adequate for recognizing occupational contact dermatitis in most of our patients (74%). PMID- 10694313 TI - Clinical evolution of alopecia areata with a male androgenetic alopecia pattern to sisaipho. AB - We report the case of a 16-year-old boy who developed an advanced alopecia areata (AA) type male androgenetic alopecia (MAGA). In 6 months he lost his frontal hair line and showed a sisaipho pattern of alopecia. It probably represents a rare evolution of AA in a wave-like form. PMID- 10694314 TI - Grzybowski's generalized eruptive keratocanthomas: a case report. AB - A 47-year old woman with Grzybowski's generalised eruptive keratoacanthomas is described. There was no history of skin disease in her family, except for an uncle's basal cell carcinoma. From 1995 she developed multiple lesions of various size, ranging from hundreds of small follicular lesions to large typical keratoacanthomas up to 5 cm in diameter, scleroderma-like facial skin and marked ectropion. Histological examination of small and large skin lesions was typical of keratoacanthoma, and no human papillomavirus was detected by polymerase chain reaction. Oral treatment with acitretin had no effect. Both cyclophosphamide and methotrexate therapy were refused by the patient despite the progressive course of the disease. Blepharoplastic surgery had some effect on eye symptoms. The etiology of this rare disease is unknown, but is probably related to some genetic defect. PMID- 10694315 TI - Extensive haemorrhagic-bullous skin manifestation of systemic AA-amyloidosis associated with IgGlambda-myeloma. AB - In an 86-year-old woman with a multiple myeloma of the IgG lambda subtype a coinciding systemic amyloidosis manifested as a macroglossia, diffuse alopecia and generalized cutaneous involvement. The skin was affected by milium-like papules, petechial haemorrhages and an increased tissue fragility with subsequent blister formation. The typical histology and immunohistology pattern revealed large intradermal amyloid masses, reacting positively with anti-amyloid A antibodies, which surrounded cuff-like dilatated blood capillaries. The abundance of these amyloid deposits led to significant deflexibilization and fragility of the capillaries and the dermal matrix eventually resulting in the haemorrhagic bullous eruptions. The peculiar feature of the present case is the intensity of bullous-haemorrhagic skin damage due to amyloid A deposition without any detection of cutaneous IgGl as the myeloma-derived paraprotein assumed to be causative for the development of systemic AA amyloidosis. PMID- 10694316 TI - Early detection of lymph node metastases by 7.5 MHz-ultrasound examination in a patient with primary malignant melanoma of the lung. AB - Primary malignant melanoma of the lung (PMML) is a rare neoplasm that may be misdiagnosed as one of the more common types of lung cancer. Most cases are characterized by a very poor prognosis, ultimately leading to the patient's death. Since an optimal systemic treatment schedule is not established so far, early detection of lymph node metastases may be important for surgical interventions. We report on a 55-year-old male patient with a primary bronchial malignant melanoma of his left lower lobe that was treated by pneumonectomy. 11 and 17 months after removal of the PMML, suspicious lymph nodes in the patient's left axilla were identified by 7.5 MHz-ultrasound examinations. Again surgical treatment was performed and histopathology showed lymph node metastases of malignant melanoma. During adjuvant therapy with interferon alpha (3 x 6 Mio IE per week) no further relapse has been observed with a follow-up of 8 months after the last operation. An overview of primary melanoma of the lung is given and diagnostic options are discussed. The 7.5 MHz-ultrasound examination appears to be especially helpful in the early detection of lymph node metastases leading to early initiation of surgical treatment possibly associated with a prolonged survival in our patient. PMID- 10694317 TI - Nail alterations secondary to pactitaxel [corrected] therapy. AB - Docetaxel and pactitaxel [corrected] are the main drugs pertaining to the taxanes family. Nail alterations associated with docetaxel therapy are not a rare event, but they have rarely been reported secondary to paclitaxel therapy, probably because of its more recent use. We present two cases of onycholysis and nail discoloration secondary to paclitaxel therapy. PMID- 10694318 TI - A case of persistent light reaction phenomenon to ketoprofen? PMID- 10694320 TI - European Dermatology Forum Reporter June 1999 including highlights of the second general meeting... PMID- 10694319 TI - The use of systemic antimycotics in dermatotherapy. AB - Fungal infections of the skin as well as of the nails and hair due to dermatophytes or due to yeasts or moulds still form a major portion of skin diseases overall. Effective therapy of mycoses is not always simple to achieve. In less severe cases topical therapy can be sufficient, but in extensive cutaneous infections, previous resistance to treatment and especially hyperkeratotic tinea and onychomycosis, systemic therapy can be mandatory. For systemic therapy, in particular azoles, i.e. itraconazole and fluconazole as well as the allylamine terbinafine are worth considering. The older antimycotics, i.e. griseofulvin and also ketoconazole are more and more replaced by other, newer drugs. For optimal treatment of a given mycosis, therapy can and should correspond to the individual situation. This applies both to the type of drug and its mode of application. The treatment of choice is the one with the best benefit to risk ratio and the best benefit to cost ratio. Unfortunately, as yet, a cure cannot be expected in every single case. PMID- 10694321 TI - Papillomavirus detection: demographic and behavioral characteristics influencing the identification of cervical disease. AB - OBJECTIVE: This study was undertaken to assess the association between detection of high-risk types of human papillomavirus and various demographic and behavioral characteristics and to further relate this association to cervical histopathologic findings. STUDY DESIGN: A total of 1007 patients with a Papanicolaou test result reported as high-grade squamous intraepithelial lesion or with 2 results reported as atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesion were referred from city and county clinics to a colposcopic clinic. All women had a cervical smear obtained, underwent colposcopically directed biopsy and endocervical curettage, and had a specimen taken for human papillomavirus deoxyribonucleic acid detection by polymerase chain reaction. Demographic information was obtained from each patient. RESULTS: Human papillomavirus deoxyribonucleic acid was identified in 655 (66%) of the specimens. High-risk human papillomavirus types (16, 18, 31, 33, and 35) were detected in 463 (70.7%) of these specimens. The prevalence of evidence of human papillomavirus (koilocytosis) and grade 1 cervical intraepithelial neoplasia in the biopsy specimen decreased significantly with age, whereas the prevalence of grade 2 or 3 cervical intraepithelial neoplasia in the biopsy specimen increased with age. There was a significant age-dependent decreasing trend in detection of high-risk human papillomavirus deoxyribonucleic acid among women who had human papillomavirus-associated changes, grade 1 cervical intraepithelial neoplasia, and grade 2 or 3 cervical intraepithelial neoplasia in the biopsy specimen. The prevalences of high-risk human papillomavirus among patients with grade 1 cervical intraepithelial neoplasia and grade 2 or 3 cervical intraepithelial neoplasia were similar, and both were significantly higher than among women with no evidence of cervical intraepithelial neoplasia or koilocytosis in the biopsy specimen. Risk factors associated with grade 2 or 3 cervical intraepithelial neoplasia were different from those associated with human papillomavirus-associated changes and with grade 1 cervical intraepithelial neoplasia. CONCLUSION: The detection of high-risk human papillomavirus was age-dependent for all histologic categories. Patients with grade 2 or 3 cervical intraepithelial neoplasia had a prevalence of high risk human papillomavirus that was similar to that among women with grade 1 cervical intraepithelial neoplasia but significantly higher than that among women whose biopsy specimens appeared normal or demonstrated only the presence of human papillomavirus-induced changes (koilocytosis). This suggests that separation of human papillomavirus-associated changes only from grade 1 cervical intraepithelial neoplasia may be of significance in tissue diagnosis. PMID- 10694323 TI - Adherence to postmenopausal hormone therapy during the year after the initial prescription: A population-based study. AB - OBJECTIVE: In randomized trials a higher proportion of women prescribed continuous combined hormone replacement therapy complete the full course of treatment compared with those prescribed sequential therapy. We sought to determine adherence to hormone therapy in a less-selected population. STUDY DESIGN: Women enrolled in a prepaid health plan participated in a telephone interview 12 to 15 months after newly initiating use of hormone replacement therapy. The interview elicited information on whether the women were still taking the hormones as first prescribed and reasons for switching or discontinuation. A computerized pharmacy database was used to determine initial doses, prescription renewal, and dates of switching or discontinuation. RESULTS: The proportion continuing the originally prescribed hormone regimen at 1 year was higher among continuous combined therapy users (68.9%, 62/90) than among sequential therapy users (54.4%, 62/114). Women who initiated continuous combined therapy were less likely to have switched regimens (10.0%) than were sequential users (20.2%; relative risk, 0.5; 95% confidence interval, 0.2-1.0) but only somewhat less likely to have discontinued use (21.2% vs 25.4%; relative risk, 0.7; 95% confidence interval, 0.4-1.3). Examined as a whole, women prescribed continuous combined therapy were less likely than those prescribed sequential therapy to quit or switch during the first year (relative risk, 0.6; 95% confidence interval, 0.4-1.0). CONCLUSION: Although adherence was higher among women prescribed continuous combined hormone replacement therapy than sequential therapy, the high level of nonadherence in both groups suggests room for improvement of menopausal therapies so that women find them acceptable for sustained use. PMID- 10694322 TI - Oral contraceptives and bone mineral density: A population-based study. AB - OBJECTIVE: We sought to test the hypothesis that exposure to oral contraceptives protects the skeleton. STUDY DESIGN: Multiple regression techniques were used to analyze data for a random sample of 710 Australian women (age range, 20-69 years). Bone mineral density was measured at the lumbar spine, proximal femur, whole body, and distal forearm. Oral contraceptive exposure was assessed by a questionnaire. RESULTS: Women exposed to oral contraceptives had a 3.3% greater mean bone mineral density adjusted for body mass index and age at the lumbar spine (partial r (2) = 0.009; P =.014). Adjusted mean vertebral bone mineral density was 3.3% greater for premenopausal women (partial r (2) = 0.008; P <.05), but the effect did not reach significance among postmenopausal women. Higher bone mineral density was associated with increased duration of exposure, with a mean increase of 3.2% associated with the first 5 years and a further 0.2% with >/=5 years of exposure. No association was detected at other sites. CONCLUSION: Exposure to oral contraceptives may be associated with higher lumbar spine bone mineral density. PMID- 10694324 TI - Modulation of the sleep electroencephalogram by estrogen replacement in postmenopausal women. AB - OBJECTIVE: We performed an examination of the effects of estrogen replacement on the sleep electroencephalogram in postmenopausal women. STUDY DESIGN: A sleep electroencephalogram was recorded in 11 postmenopausal women with and without estrogen administered by skin patch (50 microg of estradiol per day). RESULTS: Estrogen enhanced rapid-eye-movement sleep (50 +/- 4 vs 39 +/- 5 minutes, P <.05) and reduced time awake (12 +/- 5 vs 20 +/- 6 minutes, P <.05) during the first 2 sleep cycles. The normal decrease in slow-wave sleep and delta activity from the first to the second cycle (in percentage from the first cycle) was restored by estrogen (-56% +/- 9% vs -5% +/- 14% and -20% +/- 6% vs -2% +/- 5%; P <.05, respectively). Sigma electroencephalographic activity was increased by estrogen from the first to the second half of the night but decreased during baseline. CONCLUSION: Estrogen treatment after menopause can help to restore the normal sleep electroencephalogram pattern, which in turn might contribute to improved cognitive functioning. PMID- 10694325 TI - Interleukin 1 receptor antagonist gene polymorphism in women with vulvar vestibulitis. AB - OBJECTIVE: Vulvar vestibulitis is a chronic inflammatory syndrome of unknown cause and pathogenesis. We examined the relation between vulvar vestibulitis and polymorphisms in the gene coding for the interleukin 1 receptor antagonist, a naturally occurring down-regulator of proinflammatory immune responses. STUDY DESIGN: Cells from the lower genital tract of 68 women with vulvar vestibulitis, 343 women with no history of vulvodynia, and 40 women with human papillomavirus cervical infection were tested by polymerase chain reaction for the different alleles of the gene encoding for interleukin 1 receptor antagonist. The presence of human papillomavirus in the specimens was determined by polymerase chain reaction with the use of degenerate consensus primers to the conserved L1 gene. RESULTS: Allele 2 of the gene encoding the interleukin 1 receptor antagonist was present in homozygous form in 52.9% of women with vulvar vestibulitis. In marked contrast only 8. 5% of the control women and 2.5% of women with human papillomavirus were homozygous for this allele (P /=4 times a week throughout pregnancy, and a group of women who initially performed endurance exercises but then stopped exercising during midpregnancy. Exercise was monitored, and longitudinal estimates of maternal total mass and fat mass were obtained. RESULTS: Tumor necrosis factor alpha levels were lower during pregnancy in the women who exercised, and the same was true for leptin levels. When women stopped exercising, however, both tumor necrosis factor alpha and leptin concentrations rose at rates comparable to those seen in the physically active control group. Changes in leptin concentration but not those in tumor necrosis factor alpha concentration correlated with the pregnancy-associated increases in total body and fat mass. CONCLUSIONS: Regular weight-bearing exercise during pregnancy suppresses the pregnancy-associated changes normally seen in both tumor necrosis factor alpha and leptin. The decrease in leptin reflects decreased fat accretion, and we speculate that the changes in tumor necrosis factor alpha may reflect a change in insulin resistance. PMID- 10694329 TI - Eclampsia. VIII. Risk factors for maternal morbidity. AB - OBJECTIVE: This study was undertaken to identify risk factors associated with adverse maternal outcome in pregnancies complicated by eclampsia. STUDY DESIGN: This was a descriptive study of 399 consecutive women with eclampsia whose cases were managed at one perinatal center between August 1977 and July 1998. Data were collected. Risk factors studied included maternal age, race, parity, preexisting medical complications, and clinical and laboratory findings. Outcome variables were maternal morbidities. Data were analyzed by either chi(2) analysis or the unpaired Student t test as appropriate. RESULTS: In the entire cohort of women with eclampsia major maternal complications included abruptio placentae (10%), HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome (11%), disseminated intravascular coagulopathy (6%), neurologic deficits and aspiration pneumonia (7%), pulmonary edema (5%), cardiopulmonary arrest (4%), acute renal failure (4%), and death (1%, n = 2 patients with antepartum onset). Women with antepartum eclampsia had significantly higher incidences of abruptio placentae (12% vs 6%; P <.05) and HELLP syndrome (14% vs 4%; P =. 005) than did those in whom eclampsia developed post partum. In contrast, women with postpartum eclampsia were more likely to have neurologic deficits develop (9% vs 2%; P =.0006) than were those with antepartum eclampsia. In addition, women in whom eclampsia developed at /=1 arteriovenous and no arterioarterial anastomoses. Birth weight discordancy correlated with placental territory discordancy (P <.0001) and the degree of balance in arteriovenous anastomoses (P =.004). The larger placental share twin had a greater growth velocity than its smaller placental share co-twin (P =.008) for all but one anastomotic pattern. Where arteriovenous anastomoses were aligned with the net venous outflow to the fetus with the smaller territory, co-twins had similar birth weights and growth velocities irrespective of placental share. Fetal survival was higher in pregnancies with an arterioarterial anastomosis (P =.01) but lower with a venovenous anastomosis (P =. 01). Survival of both fetuses was inversely associated with birth weight discordancy (P <.0001). CONCLUSION: Although interrelationships among the various types of anastomoses are complex, our data suggest that the placental territory share and the pattern of arteriovenous anastomoses influence fetal growth, that arterioarterial anastomoses protect against fetofetal transfusion syndrome, and that venovenous anastomoses reduce perinatal survival. PMID- 10694347 TI - Esophageal atresia in the Northern Region Congenital Anomaly Survey, 1985-1997: prenatal diagnosis and outcome. AB - OBJECTIVE: This study was undertaken to determine the rate of prenatal diagnosis and surgical outcome of all cases of esophageal atresia reported to the Northern Region Congenital Anomaly Survey. STUDY DESIGN: A retrospective review was conducted on maternal and infant case notes of all cases of esophageal atresia in the Northern Region from 1985-1997, inclusive. RESULTS: A total of 176 cases of esophageal atresia was reported, and 158 diagnoses were confirmed after birth. Six cases were excluded because of incomplete data. Among the 32 patients in whom esophageal atresia was suspected antenatally because of an absent stomach bubble and hydramnios, 14 (44%) had esophageal atresia confirmed postnatally. In 10 of the 18 patients with false-positive diagnoses the stomach was subsequently seen. Esophageal atresia should have been suspected prenatally in a further 38 patients with polyhydramnios, 3 of whom also had an absent stomach bubble. There were 12 pregnancy terminations, 1 spontaneous abortion, and 19 perinatal deaths (including 9 stillbirths). Among the patients with esophageal atresia, 63.2% had associated anomalies (including 5.3% with aneuploidy), and 78.4% of these anomalies were missed prenatally. Among the live births 21.5% of the infants had a birth weight below the 5th percentile. One hundred eight (90%) had esophageal atresia with a distal tracheoesophageal fistula, and overall 102 (85%) underwent a primary repair. Among the 120 infants who underwent surgical treatment 11 subsequently died, and 6 of these deaths were related to postoperative complications. Thirty-nine infants (32.5%) had postoperative gastroesophageal reflux, necessitating fundoplication in 21 cases. At 2-year follow-up 23 of 89 infants had dysphagia, for which 7 still required a gastrostomy or jejunostomy. Infants in whom the condition was diagnosed prenatally were more likely to need prolonged mechanical ventilation, to have a longer hospital stay, and to have long-term gastrointestinal problems. CONCLUSIONS: Most cases of esophageal atresia are not suspected prenatally. Among fetuses with ultrasonographic features suggestive of esophageal atresia, 50% have the disorder confirmed postnatally. Overall perinatal and infant mortality rate among those with esophageal atresia is high (21.6%), and a further 21% of affected infants have significant morbidity after the age of 2 years. PMID- 10694348 TI - The role of estrogen in the maintenance of primate pregnancy. AB - OBJECTIVE: The aim of this study was to determine the role of estrogen in pregnancy maintenance in baboons by suppressing estrogen synthesis through administration of a highly specific nonsteroidal aromatase inhibitor, CGS 20267. STUDY DESIGN: CGS 20267 was administered subcutaneously at maximal dosages of 2.0 mg/d to pregnant baboons (n = 24) daily beginning on either day 30 (n = 8), day 60 (n = 8), or day 100 (n = 8) of gestation (normal length of gestation is 184 days) until animals miscarried or were delivered abdominally on days 160 through 168 of gestation. CGS 20267 and estradiol (n = 9), each at maximal dosages of 2 mg/d, were administered at the same intervals of gestation. Twenty baboons served as untreated control animals. Serum estradiol and progesterone levels were determined by radioimmunoassay from serum samples obtained at 1- to 3-day intervals from a maternal peripheral vein. RESULTS: Within 1 to 3 days of the initiation of CGS 20267 administration, maternal serum estradiol concentration decreased to and remained at a level that was substantially lower (mean +/- SE, 0. 096 +/- 0.003 ng/mL) than in the untreated control animals throughout gestation (0.35-4.0 ng/mL; P <.001). Although pregnancy was maintained in 19 of the 20 untreated control baboons (95%), only 12 of the 24 animals that received CGS 20267 (50%) maintained pregnancy. In contrast, all the baboons treated concomitantly with estradiol and CGS 20267 (9/9) maintained pregnancy. Thus estradiol alone prevented the high rate of miscarriage induced by the antiestrogenic agent CGS 20267. Serum progesterone concentrations were not significantly different throughout the experimental period between the CGS 20267 treated baboons that maintained pregnancy (12. 9 +/- 1.4 ng/mL) and those that miscarried (13.6 +/- 1.6 ng/mL) and were not lower in antiestrogen-treated baboons than in untreated control baboons (10.6 +/- 0.8 ng/mL). CONCLUSION: Estrogen, acting directly, indirectly, or both through a factor or factors other than the level of progesterone, plays a critically important physiologic role in the maintenance of primate pregnancy. PMID- 10694349 TI - The effect of fetal fibronectin testing on admissions to a tertiary maternal fetal medicine unit and cost savings. AB - OBJECTIVE: Fetal fibronectin bedside testing has been proposed as a diagnostic tool for the accurate diagnosis of preterm labor. The study objective was to determine whether the introduction of routine fetal fibronectin bedside testing affected costs and transfer rates from referral district hospitals to a tertiary obstetric hospital, as well as direct admissions to a tertiary referral hospital. STUDY DESIGN: We performed an 18-month prospective audit of fetal fibronectin use in 9 referral hospitals and one university maternal-fetal medicine unit. Data collected were delivery details and cervical dilatation at admission. Cost savings in terms of transport costs for patients with a negative fetal fibronectin result who were not transferred or admitted to the tertiary center were calculated for interhospital transfer (road ambulance or fixed-wing retrieval). RESULTS: One hundred fifty-one patients had a presumptive diagnosis of threatened preterm labor. Forty-five patients had a positive fetal fibronectin result and 106 had a negative fetal fibronectin result (3 with cervical dilatation >/=3 cm). Eleven (24%) patients with a positive fetal fibronectin result were delivered within 7 days, and 5 (5%) with a negative fetal fibronectin result were delivered within 7 days. One patient was delivered at 34 weeks, and the remaining patients were delivered at or after 36 weeks' gestation. All 3 patients with negative fetal fibronectin results with cervical dilatation of >/=3 cm were delivered within 5 days, leaving 2 (1.9%) patients (with closed cervices and negative fetal fibronectin results) being delivered 5 days after the fetal fibronectin testing. Ninety percent of the patients admitted to a referral hospital with threatened preterm labor who had a negative fetal fibronectin result were not transferred; thus an unnecessary transfer was avoided, with cost savings ranging from $30,297 for road and fixed-wing transport. CONCLUSION: A negative fetal fibronectin result is not helpful if cervical dilatation is present, and these patients should be treated as having a high risk of preterm delivery. The use of a fetal fibronectin test was associated with a 90% reduction in maternal transfer and can substantially reduce the costs and inconvenience associated with unnecessary transfer. PMID- 10694350 TI - Low midpregnancy placental volume in rural Indian women: A cause for low birth weight? AB - OBJECTIVE: We sought to study midpregnancy placental volume in rural Indian women, its maternal determinants, and its relationship to neonatal size. STUDY DESIGN: We performed a prospective community-based study of maternal nutrition and fetal growth in 6 villages near the city of Pune. Measurements included midpregnancy placental volume determined by means of ultrasonography at 15 to 18 weeks' gestation, maternal anthropometric measurements before and during pregnancy, and maternal blood pressure and biochemical parameters during pregnancy. Neonatal size and placental weight were measured at birth. RESULTS: The mothers were short and underweight (mean height, 1.52 m; weight, 42 kg; body mass index, 18 kg/m(2)) and produced small babies (mean birth weight, 2648 g). Midpregnancy placental volume (median, 144 mL) was related to the mother's prepregnancy weight (r = 0.15; P <.001) but not to weight gain during pregnancy, blood pressure, or circulating hemoglobin, ferritin, red blood cell folate, or glucose concentrations. Midpregnancy placental volume was related to placental weight at birth (r = 0.29; P <.001) and birth weight (r = 0.25; P <.001) independent of maternal size. CONCLUSION: In Indian mothers midpregnancy placental volume is significantly associated with prepregnant maternal weight and is an independent predictor of birth weight. Our findings may provide clues to the high prevalence of low-birth-weight infants in India. PMID- 10694351 TI - Histologic and biochemical study of the brain, heart, kidney, and liver in asphyxia caused by occlusion of the umbilical cord in near-term fetal lambs. AB - OBJECTIVE: We sought to determine the relationship between the degree of histologic changes in the brain, heart, kidney, and liver in fetal lambs after severe asphyxia and to analyze the role of oxidative stress in the pathogenesis of fetal multiple organ failure. STUDY DESIGN: Eight chronically instrumented near-term fetal lambs were asphyxiated by partial umbilical cord occlusion for approximately 60 minutes until the fetal arterial pH reached <6.9 and the base excess reached <-20 mEq/L. An additional 6 fetuses were used as sham-asphyxiated controls. Fetal heart rates, blood pressure, fetal breathing movements, and arterial blood gases and acid-base states were serially monitored. The brain, heart, kidney, and liver were collected 72 hours after asphyxia, processed, and histologically examined after hematoxylin and eosin staining. Fetal brain histologic features were classified into 5 grades, with 5 being the most severe damage. The other organs were examined histologically by pathologists who were blinded to the treatment. Each organ was assayed for tissue concentrations of thiobarbituric acid-reactive substances, superoxide dismutase, glutathione, lactate, and glucose. RESULTS: Myocardial changes of necrosis, phagocytosis, and contraction bands occurred in only 2 of the most severely (grade 5) brain-damaged fetuses. The same 2 cases showed fatty changes and congestion in the liver. In the kidney all asphyxiated cases showed tubular necrosis, but glomeruli were generally spared. Of the measures of oxidative stress, only liver tissue levels of thiobarbituric acid-reactive substances and superoxide dismutase were significantly higher in the asphyxiated group than in the control group, but there was no correlation with the degree of damage. Lactate level was higher only in the heart in the asphyxiated fetuses. CONCLUSION: Renal tubular damage was seen with all degrees of asphyxia, despite variable brain damage. Histologic changes in the myocardium and liver were seen only with the most severe brain damage. Oxidative stress appears to play a role in the pathogenesis of liver damage. PMID- 10694352 TI - Reduction-oxidation (redox) state regulation of matrix metalloproteinase activity in human fetal membranes. AB - OBJECTIVE: The mechanisms underlying membrane rupture at term and preterm are obscure. Collagenolytic activity of matrix metalloproteinases in amniochorionic membranes increases during spontaneous term and preterm labor associated with intra-amniotic infection. We sought to test the hypothesis that reduction oxidation homeostasis, which is altered in inflammatory states, directly regulates amniochorionic matrix metalloproteinases. STUDY DESIGN: Membranes were collected from 7 patients undergoing elective cesarean delivery at term, rinsed thoroughly, and immediately incubated in phosphate-buffered sodium chloride solution at 37 degrees C for 24 hours. Matrix metalloproteinase activity in the culture medium was assayed by substrate-gel electrophoresis and normalized against the dry weight of the tissue incubated. Superoxide anions were generated in the presence of membranes by a xanthine (2 mmol/L) and xanthine oxidase (20 mU/mL) mixture and monitored by reduction of ferri-cytochrome c to ferro cytochrome c. Incubations were performed in the presence of xanthine alone, a xanthine-xanthine oxidase mixture, superoxide dismutase (500 U/mL), a xanthine xanthine oxidase-superoxide dismutase mixture, nitro-L-arginine (a nitric oxide synthase inhibitor, 1 mmol/L), xanthine-xanthine oxidase-nitro-L-arginine, S nitroso-N -acetylpenicillamine (a nitric oxide donor, 10 mmol/L), xanthine xanthine oxidase-S-nitroso-N -acetylpenicillamine, N -acetylcysteine (a thiol containing antioxidant, 0.1, 1, or 10 mmol/L), lipopolysaccharide (100 ng/mL), or lipopolysaccharide-N -acetylcysteine. Intracellular generation of superoxide anions was monitored by the reduction of nitroblue tetrazolium to formazan. RESULTS: Basal matrix metalloproteinase 9 and matrix metalloproteinase 2 levels were detected in all samples. Superoxide anions significantly increased matrix metalloproteinase 9 activity but did not increase matrix metalloproteinase 2 activity, which effect was reversed by the addition of superoxide dismutase. N acetylcysteine reduced basal activity of both matrix metalloproteinase 9 and matrix metalloproteinase 2 to 20%. Importantly, N-acetylcysteine completely inhibited intracellular formazan formation in cultured membranes both in the absence and in the presence of lipopolysaccharide. Neither nitric oxide synthase inhibition nor the nitric oxide donor S-nitroso-N -acetylpenicillamine had any effect on fetal membrane matrix metalloproteinase activity. CONCLUSION: Matrix metalloproteinase activity in human fetal membranes is reduction-oxidation (redox)-regulated. Matrix metalloproteinase 9 activity in human fetal membranes is directly increased by superoxide anion, a byproduct of macrophages and neutrophils. Neither nitric oxide donors nor nitric oxide synthase inhibitors significantly affect matrix metalloproteinase activity in human fetal membranes. The glutathione precursor N-acetylcysteine dramatically inhibits amniochorionic matrix metalloproteinase activity in addition to inhibiting intrinsic superoxide generation within the tissue. Thus thiol-reducing agents, such as N acetylcysteine, may be beneficial in preventing preterm premature rupture of the membranes. PMID- 10694353 TI - A systematic review and meta-analysis of prospective studies on the association between maternal cigarette smoking and preterm delivery. AB - We have attempted to quantify the most up-to-date estimate of the association between cigarette smoking by the mother and preterm delivery. Studies were selected for inclusion in this review if they were prospective, reported data stratified across at least two levels of maternal smoking, and defined preterm delivery on the basis of gestational age. In a meta-analysis we combined results from multiple studies that reported on preterm delivery and maternal smoking during pregnancy. Pooled odds ratios were computed for various strata of smoking intensity with the Mantel-Haenszel fixed-effects model. Twenty studies met all inclusion criteria and were included in meta-analysis. The pooled point estimate from 20 prospective studies on any maternal smoking versus no maternal smoking was 1.27 (95% confidence interval, 1.21-1.33). Subgroup analyses stratifying maternal smoking on number of cigarettes per day suggest a dose-response relationship at low to moderate levels of smoking, which was not further increased at high levels of smoking. A nonsignificant level of publication bias appears to exist in the smoking-preterm delivery literature. Cigarette smoking is a preventable risk factor that is associated with preterm delivery. Consistent results across many study populations and research designs and evidence of a dose response relationship support its causal role in preterm delivery. PMID- 10694354 TI - Failure of initiation of parturition after electrocoagulation of the pituitary of the fetal lamb. American Journal of Obstetrics and Gynecology, volume 98, app. 1080-1086, 1967. PMID- 10694355 TI - The role of the hypothalamic-pituitary-adrenal axis in preparing the fetus for birth. PMID- 10694356 TI - Safety of multiple courses of corticosteroid treatment. PMID- 10694358 TI - Value of human papillomavirus testing. PMID- 10694360 TI - Ethics committee approval of published study. PMID- 10694362 TI - Intrauterine insemination--how far should we go? PMID- 10694364 TI - The role of the StAR protein in steroidogenesis: challenges for the future. AB - The steroidogenic acute regulatory or StAR protein has been shown to be instrumental in the acute regulation of steroid hormone biosynthesis through its action in mediating cholesterol transfer to the inner mitochondrial membrane and the cholesterol side chain cleavage enzyme system. Since the time of its cloning in 1994, a number of studies have been performed which underscore the important role that this protein plays in steroidogenesis. While it is now quite apparent that StAR fulfills the criteria for the acute regulator as proposed by early studies, several crucial areas remain poorly understood. This list is topped by the so far intractable nature of the mechanism of action of StAR in transferring cholesterol to the P450scc enzyme. A second area which should prove to be of great interest is that of further understanding the regulation of the StAR gene which, like many genes, is quite complex. Lastly, with the recent demonstration of StAR being present in the brain, determining if StAR has a role in the synthesis of neurosteroids should prove to be of great importance. PMID- 10694365 TI - ACTH treatment disrupts ovarian IGF-I and steroid hormone production. AB - Hyper-adrenal activity and increased glucocorticoid hormone release are associated with disruptions in reproductive function and adverse effects on the ovary. The aim of this investigation was to determine whether elevated glucocorticoid hormone levels can influence ovarian IGF-I synthesis and action in vivo. To elevate endogenous glucocorticoid levels, gilts were treated with ACTH during the luteal phase of the oestrous cycle (days 9-13) while the control group received saline. The gilts were subsequently ovariectomized on either day 14 or day 18 of the oestrous cycle. Follicular fluid (FF) was collected from individual follicles; IGF-I and steroid hormone concentrations were determined by radioimmunoassay, and IGF-binding protein (IGFBP) expression was assessed by Western ligand blotting. Granulosa cells were also recovered and placed in culture to determine IGF-I, progesterone (P(4)) and oestradiol-17beta (E(2)) production levels. The cells were cultured in serum-free medium for 5 days and supplemented with: (a) media alone, (b) IGF-I, (c) FSH and androstenedione (A(4)), or (d) IGF-I with FSH and A(4). The FF from ACTH-treated gilts was characterized by elevated (P<0.05) cortisol levels on day 14 and lower (P<0.05) E(2) values on both day 14 and day 18. Lower (P<0.05) IGF-I concentrations were also measured in the FF of ACTH-treated gilts collected on day 18. This altered hormone profile in FF was associated with impaired IGF-I and steroid hormone synthesis by granulosa cells. IGF-stimulated P(4) production (P<0.01) by cells recovered from ACTH-treated gilts on day 14 was lower (P<0.05). By day 18, IGF-I, P(4) and E(2) production by cells from the ACTH group were all significantly (P<0. 05) lower. These results demonstrate that increased glucocorticoid concentrations can disrupt subsequent ovarian IGF-I synthesis and IGF action in vivo and can, potentially, impair follicle maturation. PMID- 10694366 TI - Effect of neonatal and adult testosterone treatment on the cellular composition of the adult female rat anterior pituitary. AB - The adult female pituitary has significantly more lactotrophs than that of the male, while the later has a higher percent of somatotrophs. It is clear that GH and prolactin (PRL) gene expression and somatotroph and lactotroph proliferation are modulated by the postpubertal hormone environment; however, the role of the neonatal steroid environment in this process is not known. We have used in situ hybridization to determine the number of GH and PRL mRNA-containing cells, as well as the level of expression of these two hormones, in response to neonatal and adult testosterone treatment. Female rats exposed to testosterone during the neonatal period, adulthood or both periods, as well as normal females and males were used. Exposure to testosterone during the neonatal period significantly increased the percentage of somatotrophs (ANOVA: P<0. 005) and decreased that of lactotrophs in the adult female rat (ANOVA: P<0.001). Adult testosterone treatment had no significant effect on the percentage of somatotrophs. The percentage of lactotrophs was significantly increased by adult testosterone only in those rats also exposed to neonatal testosterone. PRL mRNA concentrations, as reflected by silver grains/cell, were reduced by neonatal testosterone and increased by adult testosterone treatment (ANOVA: P<0.0001). Overall PRL mRNA levels, measured by densitometry, were also reduced by neonatal testosterone exposure, but adult testosterone had no effect (ANOVA: P<0.001). GH mRNA levels per cell, as reflected by silver grains/cell, were increased by adult testosterone, while neonatal testosterone treatment had no effect. Overall GH mRNA levels per unit area, determined by densitometry measurements, were increased by both neonatal and adult testosterone treatment, with the combination of these two treatments resulting in adult females having levels indistinguishable from intact males (ANOVA: P<0.003). These results suggest that, in combination with postpubertal sex steroids, the neonatal gonadal steroid environment plays an important role in determining anterior pituitary hormone synthesis and cellular composition. PMID- 10694367 TI - Prolactin induction of insulin gene expression: the roles of glucose and glucose transporter-2. AB - Previous studies have shown that lactogenic hormones stimulate beta-cell proliferation and insulin production in pancreatic islets. However, all such studies have been conducted in cells incubated in medium containing glucose. Since glucose independently stimulates beta-cell replication and insulin production, it is unclear whether the effects of prolactin (PRL) on insulin gene expression are exerted directly or through the uptake and/or metabolism of glucose. We examined the interactions between glucose and PRL in the regulation of insulin gene transcription and the expression of glucose transporter-2 (glut 2) and glucokinase mRNAs in rat insulinoma (INS-1) cells. In the presence of 5.5 mM glucose, the levels of preproinsulin and glut-2 mRNAs in PRL-treated cells exceeded the levels in control cells (1.7-fold, P<0.05 and 2-fold, P<0.05 respectively). The maximal effects of PRL were noted at 24-48 h of incubation. PRL had no effect on the levels of glucokinase mRNA. The higher levels of glut-2 mRNA were accompanied by an increase in the number of cellular glucose transporters, as demonstrated by a 1. 4- to 2.4-fold increase in the uptake of 2 deoxy-d-[(3)H]glucose in PRL-treated INS-1 cells (P<0.001). These findings suggested that the insulinotropic effect of PRL is mediated, in part, by induction of glucose transport and/or glucose metabolism. Nevertheless, even in the absence of glucose, PRL stimulated increases in the levels of preproinsulin mRNA (3.4-fold higher than controls, P<0.0001) and glut-2 mRNA (2-fold higher than controls, P<0.01). These observations suggested that PRL exerts glucose independent as well as glucose-dependent effects on insulin gene expression. Support for this hypothesis was provided by studies of insulin gene transcription using INS-1 cells transfected with a plasmid containing the rat insulin 1 promoter linked to a luciferase reporter gene. Glucose and PRL, alone and in combination, stimulated increases in cellular luciferase activity. The relative potencies of glucose (5.5 mM) alone, PRL alone, and glucose plus PRL in combination were 2.2 (P<0.001), 3.4 (P<0.01), and 7.9 (P<0.0001) respectively. Our findings suggest that glucose and PRL act synergistically to induce insulin gene transcription. PMID- 10694368 TI - Two free isoforms of ovine glycoprotein hormone alpha-subunit strongly differ in their ability to stimulate prolactin release from foetal pituitaries. AB - alpha-Subunit dissociated from glycoprotein hormones has been previously shown to stimulate rat pituitary lactotroph differentiation and proliferation. However, whether the free form of the alpha-subunit (free alpha) can also play such a role is not known. To test whether free alpha may act on prolactin (PRL) release from ovine foetal pituitaries, this molecule was purified and two major isoforms, alphaA and alphaB were isolated. Free alphaA was found to be more acidic and more hydrophobic than both free alphaB and ovine LH alpha-subunit (oLHalpha). Free alphaA and oLHalpha exhibited a molecular mass of 14 kDa as determined by mass spectrometry, whereas free alphaB displayed a molecular mass of only 13.5 kDa because of its truncated N-terminus. All three alpha molecules bear mature-type N linked saccharide chains including Nacetyl galactosamine residues but none of them contains O-linked oligosaccharide. The free alphaA isoform, more than the oLHalpha, was able to stimulate PRL release from ovine foetal pituitary explants in culture, whereas the free alphaB isoform displayed no activity. Moreover, the free alphaA and alphaB isoforms were able to recombine with the ovine LH beta subunit (oLHbeta). The free alphaB/oLHbeta, and the oLHalpha/oLHbeta dimer were 4 fold more active than the free alphaA/oLHbeta dimer in a specific LH radioreceptor assay and in the stimulation of testosterone release from rat Leydig cells. The present study demonstrates that the two free alpha isoforms of ovine glycoprotein hormones exhibit distinct efficiencies in stimulating PRL release from ovine foetal pituitaries. Moreover, despite their identical ability to recombine with the oLHbeta, the free alpha isoform, which is the most efficient on PRL release, is the least efficient in conferring LH activity on the alpha/beta dimer. PMID- 10694369 TI - hCGbeta core fragment is a metabolite of hCG: evidence from infusion of recombinant hCG. AB - The availability of recombinant human chorionic gonadotrophin (r-hCG) has allowed us to measure its metabolic and renal clearance rates and to study the origin of the beta core fragment of hCG (hCGbetacf). Serum and urine samples were collected from six subjects, after an intravenous injection of 2 mg (equivalent to 44 000 IU Urinary hCG) r-hCG, and assayed for hCG and the beta subunit (hCGbeta). Urine from four of the subjects was also subjected to gel chromatography and assayed for hCGbetacf and hCG. r-hCG, administered as an intravenous dose, was distributed, initially in a volume of 3.4+/-0.7 l (mean+/-s.d.) and then in 6.5+/ 1.15 l at steady-state. The disappearance of r-hCG from serum was bi-exponential, with an initial half-life of 4.5+/-0.7 h and a terminal half-life of 29.0+/-4.6 h. The mean residence time was 28. 6+/- 3.6 h and the total systemic clearance rate of r-hCG was 226+/-18 ml/h. The renal clearance rate was 28.75+/-6.2 ml/h (mean+/-s.d). hCGbetacf was detected in all urine samples collected at 6 h intervals. Over the 138 h period of urine collection, 12.9% (range 10.1-17.3% ) of r-hCG injected was recovered as the intact molecule and 1.7% (range 0.8-2.9%) was recovered as the hCGbetacf, in 4 subjects. The molar ratio of hCGbetacf to hCG in urine increased from 3.1+/-1.7%, on day 1, to 76+/-34.3% (mean+/-s.e.m.) on day 5, after r-hCG infusion, suggesting that hCGbetacf is a metabolic product of the infused r-hCG. PMID- 10694370 TI - Stable overexpression of the glucose-6-phosphatase catalytic subunit attenuates glucose sensitivity of insulin secretion from a mouse pancreatic beta-cell line. AB - Glucose-6-phosphatase (G-6-Pase) hydrolyzes glucose-6-phosphate to glucose, reciprocal with the so-called glucose sensor, glucokinase, in pancreatic beta cells. To study the role of G-6-Pase in glucose-stimulated insulin secretion from beta cells, we have introduced rat G-6-Pase catalytic subunit cDNA and have established permanent clones with 3-, 7- and 24-fold G-6-Pase activity of the mouse beta-cell line, MIN6. In these clones, glucose usage and ATP production in the presence of 5.5 or 25 mM glucose were reduced, and glucose-stimulated insulin secretion was decreased in proportion to the increased G-6-Pase activity. In addition, insulin secretory capacity in response to d-fructose and pyruvate was unchanged; however, 25 mM glucose-stimulated insulin secretion and intracellular calcium response were completely inhibited. In the clone with 24-fold G-6-Pase activity, changes in intracellular NAD(P)H autofluorescence in response to 25 mM glucose were reduced, but the changes with 20 mM fructose and 20 mM pyruvate were not altered. Stable overexpression of G-6-Pase in beta cells resulted in attenuation of the overall glucose-stimulated metabolic responses corresponding to the degree of overexpression. This particular experimental manipulation shows that the possibility exists of modulating glucose-stimulated insulin release by thoroughly altering glucose cycling at the glucokinase/G-6-Pase step. PMID- 10694371 TI - Transmembrane protein tyrosine phosphatase IA-2 (ICA512) is expressed in human midgut carcinoids but is not detectable in normal enterochromaffin cells. AB - A potential upregulation of receptor type protein tyrosine phosphatase IA-2 (ICA512) expression was detected by differential display and investigated in midgut carcinoid tumours. Normal intestine tissue and tumour tissue from 13 midgut carcinoid patients were studied by in situ hybridisation using an IA-2 ribonucleotide probe and confocal microscopy using specific IA-2 antibodies. Previously, it had been shown that IA-2 is located in the secretory granules of virtually all neuroendocrine cells. However, we found that IA-2 was not detectable in resting normal enterochromaffin (EC) cells of the small intestine, while high expression of IA-2 mRNA and protein was confirmed in both primary and metastatic carcinoid tissue. This difference in expression was not observed with chromogranin A or serotonin, two secretory granule hormones known to be expressed in EC cells, indicating that IA-2 was seemingly not necessary for the basal production and packaging of these hormones. When comparing patients receiving biotherapy before operation with untreated patients, we found expression of IA-2 to be lower in tumours from patients that had been treated with a combination of alpha-interferon and the somatostatin analogue, octreotide. There was no correlation between IA-2 expression and proliferation rates as measured by immunohistochemistry with antibodies against the Ki 67 antigen. Furthermore, we show that IA-2 is co-localised with serotonin in carcinoid tumours as well as in the pancreatic tumour cell line, BON1, which is interesting as serotonin secretion rate is presumably higher in tumour cells than in resting EC cells. Taken together, these findings may indicate a role for IA-2 in the later stages of the regulated secretory process. PMID- 10694372 TI - Apoptosis during goitre involution - the role of Bcl-2. AB - Goitrogenesis is accompanied by hyperplasia and hypertrophy and involves tissue remodelling and angiogenesis. During the involution of the goitre there must be removal of this increased thyroid volume, in addition to further remodelling, which may involve apoptosis. We investigated apoptosis in the involuting rat thyroid using male Fisher rats that were on a goitrogenic regimen for 14 days and then returned to a normal diet. Thyroid weights increased fourfold with the goitrogenic regimen. During involution, the largest decrease in weight was between day 2 and day 4 after withdrawal of treatment. After 34 days of involution, the thyroid weight plateaued, but had not returned to control values. High levels of Bcl-2 immunoreactivity were observed in normal and goitrous rat thyroids. These high levels were significantly reduced at 2 days of involution, after which high levels of Bcl-2 immunoreactivity returned. In situ end-labelling of apoptotic cells showed that there was an increase in the number of cells undergoing DNA fragmentation during goitrogenesis (1.0+/-0.8 cells/100 cells, n=9) compared with controls, in which no positive staining was observed. After 2 days of goitrogen withdrawal, there was a further fourfold increase in the number of in situ end-labelled cells (day 16: 4.1+/-1.7, n=9). Numbers of positive cells returned to low levels after 4 days of involution (day 18: 0.3+/-0.8, n=9). Using antiserum to apoptosis-specific protein, we found increased immunoreactivity during goitrogenesis and after 2 days of involution that was localised predominantly with the stromal and vascular tissue at both time points. The data show that rapid downregulation of Bcl-2 accompanies thyroid involution, which involves increased levels of apoptosis within the stromal compartment. PMID- 10694373 TI - Modulation of uncoupling protein 2 and uncoupling protein 3: regulation by denervation, leptin and retinoic acid treatment. AB - We recently reported that the leptin-induced increase in uncoupling protein 1 (UCP1) mRNA in brown adipose tissue (BAT) is prevented by the denervation of BAT. We also reported that retinoic acid (RA) increases UCP1 mRNA in BAT. To extend these finding to UCP2 and UCP3 in BAT, we examined UCP2 and UCP3 mRNA after unilateral denervation of BAT, as well as after leptin, beta(3)-adrenergic agonist, RA, and glucocorticoid administration to rats. UCP3 mRNA was 20% less in the denervated compared with the intact BAT, whereas UCP2 mRNA was unchanged with denervation. The beta(3)-adrenergic agonist, CGP-12177 (0.75 mg/kg), increased UPC3 mRNA by 40% in the innervated and by 85% in the denervated BAT. Leptin (0.9 mg/day for 3 days) increased both UCP2 and UCP3 mRNA by 30% in the innervated and, surprisingly, in the denervated BAT. RA (7.5 mg/kg) increased UCP1 mRNA but decreased UCP2 and UCP3 mRNA by 50%, whereas methylprednisolone (65 mg/kg, two doses 24 h apart) suppressed all three uncoupling proteins by greater than 60%. The present findings indicate that: sympathetic innervation is necessary to maintain basal levels of UCP3 mRNA; beta(3)-adrenergic agonist stimulation induces UCP3 mRNA; leptin induces UCP2 and UCP3 mRNA and this induction is not dependent on sympathetic innervation; RA increases UCP1 but decreases UCP2 and UCP3 mRNA; and methylprednisolone suppresses UCP1, UCP2, and UCP3 mRNA equally. These data suggest that there are distinct patterns of regulation between UCP1, UCP2, and UCP3, and there may be at least two modes by which leptin could modulate thermogenesis in BAT; first, by increasing sympathetic stimulation of BAT and induction of UCP1 mRNA and, secondly, by increasing UCP2 and UCP3 mRNA by a mechanism independent of sympathetic stimulation. PMID- 10694374 TI - Regulation of vitamin D-1alpha-hydroxylase and -24-hydroxylase expression by dexamethasone in mouse kidney. AB - We investigated the effects of dexamethasone on vitamin D-1alpha-hydroxylase and 24-hydroxylase expression and on vitamin D receptor (VDR) content in the kidneys of mice fed either a normal (NCD) diet or a calcium- and vitamin D-deficient (LCD) diet for 2 weeks. For the last 5 days mice received either vehicle or dexamethasone (2 mg/kg per day s.c.). Dexamethasone significantly increased plasma calcium concentrations without changing plasma concentrations of 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) in both NCD and LCD groups. Northern blot and enzyme activity analyses in NCD mice revealed that dexamethasone increased renal VDR mRNA expression modestly and greatly increased 24-hydroxylase mRNA abundance and enzyme activity, but did not affect 1alpha-hydroxylase mRNA abundance and enzyme activity. In mice fed an LCD diet, dexamethasone increased renal VDR mRNA expression 1.5-fold, decreased 1alpha-hydroxylase mRNA abundance (52%) and activity (34%), and markedly increased 24-hydroxylase mRNA abundance (16-fold) and enzyme activity (9-fold). Dexamethasone treatment did not alter functional VDR number (B(max) 125-141 fmol/mg protein) or ligand affinity (K(d) 0.13-0.10 nM) in LCD mice. Subcutaneous injections of 1,25(OH)(2)D(3) (0.24 nmol/kg per day for 5 days) into NCD mice strongly increased renal 24-hydroxylase mRNA abundance and enzyme activity, while there was no effect of dexamethasone on renal 24-hydroxylase expression in these mice. This may be due to overwhelming induction of 24-hydroxylase by 1,25(OH)(2)D(3). These findings suggest that glucocorticoid-induced osteoporosis is caused by direct action of the steroids on bone, and the regulatory effect of glucocorticoids on renal 25-hydroxyvitamin D(3) metabolism may be less implicated in the initiation and progression of the disease. PMID- 10694375 TI - Neonatal handling permanently alters hypothalamic-pituitary- adrenal axis function, behaviour, and body weight in boars. AB - Neonatal handling permanently alters hypothalamic- pituitary-adrenal axis (HPA) function in rats. In the rat, this treatment increases hippocampal glucocorticoid receptors (GR) and dampens plasma ACTH and corticosterone responses to stressors. The objectives of this study were to determine whether neonatal handling of pigs would effect permanent changes in plasma corticosteroid binding capacity (CBG), basal or stressor-induced plasma cortisol and ACTH concentrations, brain or pituitary GR levels, dexamethasone suppression of plasma cortisol and ACTH concentrations, behaviour in an open field-test pen, and body weights. Twelve litters of pigs were randomly assigned to either neonatal handling or no disturbance. Handled litters were removed from the farrowing crate for 10 min per day for the first 14 days of life. Male pigs were kept for the study and the boars were weighed monthly. At 7 months of age, boars were tested for locomotory behaviour in an open field-test pen. The boars were implanted with indwelling ear vein catheters and blood samples were obtained basally, during and after application of a nose snare, and after 0.04 mg/kg dexamethasone. Boars were killed and blood samples were obtained and the brain and pituitary glands collected. Handled boars had greater (P<0.05) plasma CBG binding and lower basal total (P<0.05) and calculated free (P<0.03) plasma cortisol concentrations. No significant differences between treatments were found in plasma ACTH or cortisol responses to a nose-snare stressor; however, when killed, handled boars had greater (P<0.02) plasma ACTH concentrations. Handled and non-handled boars did not differ in plasma ACTH or cortisol responses to dexamethasone. There was no treatment effect on GR expression in the pituitary gland, frontal cortex, hippocampus, or hypothalamus. Behaviourally, the handled boars had higher (P<0.03) locomotor scores over inner squares and a lower (P<0.05) ratio of outer:inner squares entered in open field-tests. During the first 7 months of life, body weights were lower (P<0.004) for handled boars. In conclusion, neonatal handling permanently altered HPA function in pigs, but in a manner dissimilar to that found in the rat. These changes induced in the pig were not beneficial for commercial production with respect to body weight. PMID- 10694376 TI - IGF-I/IGFBPs system response to endotoxin challenge in sheep. AB - Endotoxin (LPS), a membrane component of gram-negative bacteria produces multiple endocrine and metabolic effects that mimic those seen in acute sepsis. It induces species-dependent alterations of the growth hormone (GH) axis that may participate in the shift of the metabolism towards catabolic events. Humans and sheep show increased GH secretion in response to LPS, as opposed to rats, which have been the most studied. The purpose of our work was to evaluate the effects in intact rams of an acute intravenous administration of a high dose of LPS on the insulin-like growth factor (IGF)-I/IGF-binding proteins (IGFBPs) system and to analyse the temporal relationship of GH axis changes with those of several hormonal and metabolic parameters such as somatostatin, cortisol, insulin, and glucose. LPS induced a late moderate decrease of total IGF-I plasma levels following a 5-h steady-state period (-26.6+/-4. 2%, P<0.05, 9 h after LPS), despite a biphasic and sustained increase of GH secretion in the same animals (2.48+/-0.39 ng/ml 2 h after LPS and 2.7+/-0.37 ng/ml 5 h after LPS vs 0.77+/ 0.10 before LPS; Briard et al. 1998a). Western ligand blot analysis in IGFBPs showed an early short-lasting increase in IGFBP-1 (188.8+/-39% P<0. 05, 3 h after LPS). No significant change was seen for either IGFBP-2, -3 or -4. We observed a marked and sustained increase in cortisol (128.18+/-7.21 ng/ml 3 h after LPS, vs 21.17+/-4.22 before LPS). Insulin also increased (27.69+/-3.90 microU/ml 3 h after LPS, vs 13.48+/-1.69 before LPS) and its burst coincided with that of IGFBP 1. Moderately decreased IGF-I and increased IGFBP-1 plasma levels contrasted with the sustained increase in GH secretion that we recently described, thereby suggesting that endotoxin causes a state of resistance to GH. This may be exacerbated by reduced IGF-I bioavailability and/or action, and which may participate in the pathophysiology of the catabolic state seen in sepsis. The temporal analysis of hormone responses suggests that endotoxin-induced alterations of the IGF-I/IGFBPs system may involve the prolonged and substantial somatostatin rise that we recently demonstrated, together with an increase in glucocorticoid and cytokine as more generally assumed. PMID- 10694377 TI - Amino acid regulation of gene transcription of rat insulin-like growth factor binding protein-1. AB - To investigate the molecular mechanisms of increased transcription of the insulin like growth factor-binding protein-1 (IGFBP-1) gene in dietary protein-deprived animals, the cis-acting sequence that is involved in this regulation was analyzed. We first showed that IGFBP-1 gene transcription was up-regulated by amino acid deprivation in cultured liver cell lines: H4IIE and HuH-7. Since HuH-7 cells showed a greater increase in IGFBP-1 mRNA in response to amino acid deprivation, this cell line was used in further experiments. Using a promoter function assay, we found that up-regulation of promoter activity responding to amino acid deprivation was abolished by deleting the region between -112 and -81 bp from the cap site from the gene construct. This cis-acting region includes the insulin-responsive element (IRE) and glucocorticoid responsive element (GRE) of IGFBP-1. In summary, the present observation suggests that the 32-bp (-112 to 81) in the IGFBP-1 gene 5' promoter region is involved in the induction of the IGFBP-1 gene in response to amino acid deprivation. PMID- 10694378 TI - Simultaneous binding of two DNA duplexes to the NtrC-enhancer complex studied by two-color fluorescence cross-correlation spectroscopy. AB - The transcription activator protein NtrC (nitrogen regulatory protein C, also termed NR(I)) can catalyze the transition of Escherichia coli RNA polymerase complexed with the sigma(54) factor (RNAP x sigma(54)) from the closed complex (RNAP x sigma(54) bound at the promoter) to the open complex (melting of the promoter DNA). This process involves phosphorylation of NtrC (NtrC-P), assembly of an octameric NtrC-P complex at the enhancer DNA sequence, interaction of this complex with promoter-bound RNAP x sigma(54) via DNA looping, and hydrolysis of ATP. Here it is demonstrated by two-color fluorescence cross-correlation spectroscopy measurements of 6-carboxyfluorescein and 6-carboxy-X-rhodamine labeled DNA oligonucleotide duplexes that the NtrC-P complex can bind two DNA duplexes simultaneously. This suggests a model for the conformation of the looped intermediate that is formed between NtrC-P and RNAP. sigma(54) at the glnAp2 promoter during the activation process. PMID- 10694379 TI - Fucoidan-dependent conformational changes in annexin II tetramer. AB - Fucoidan, a sulfated fucopolysaccharide, mimics the fucosylated glycans of glycoproteins and has therefore been used as a probe for investigating the role of membrane polysaccharides in cell-cell adhesion. In the present report we have characterized the interaction of fucoidan with the Ca(2+)- and phospholipid binding protein annexin II tetramer (AIIt). AIIt bound to fucoidan with an apparent K(d) of 1.24 +/- 0.69 nM (mean +/- SD, n = 3) with a stoichiometry of 0.010 +/- 0.001 mol of fucoidan/mol of AIIt (mean +/- SD, n = 3). The binding of fucoidan to AIIt was Ca(2+)-independent. Furthermore, in the presence but not the absence of Ca(2+), the binding of fucoidan to AIIt caused a decrease in the alpha helical content from 32% to 7%. A peptide corresponding to a region of the p36 subunit of AIIt, F(306)-S(313), which contains a Cardin-Weintraub consensus sequence for heparin binding, was shown to undergo a conformational change upon fucoidan binding. This suggests that heparin and fucoidan bound to this region of AIIt. The binding of fucoidan but not heparin by AIIt also inhibited the ability of AIIt to bind to and aggregate phospholipid liposomes. These results suggest that the binding of AIIt to the carbohydrate conjugates of certain membrane glycoproteins may have profound effects on the structure and biological activity of AIIt. PMID- 10694380 TI - Two homologues encoding human UDP-glucose:glycoprotein glucosyltransferase differ in mRNA expression and enzymatic activity. AB - UDP-glucose:glycoprotein glucosyltransferase (UGT) is a soluble protein of the endoplasmic reticulum (ER) that operates as a gatekeeper for quality control by preventing transport of improperly folded glycoproteins out of the ER. We report the isolation of two cDNAs encoding human UDP-glucose:glycoprotein glucosyltransferase homologues. HUGT1 encodes a 1555 amino acid polypeptide that, upon cleavage of an N-terminal signal peptide, is predicted to produce a soluble 173 kDa protein with the ER retrieval signal REEL. HUGT2 encodes a 1516 amino acid polypeptide that also contains a signal peptide and the ER retrieval signal HDEL. HUGT1 shares 55% identity with HUGT2 and 31-45% identity with Drosophila, Caenorhabditis elegans, and Schizosaccharomyces pombe homologues, with most extensive conservation of residues in the carboxy-terminal fifth of the protein, the proposed catalytic domain. HUGT1 is expressed as multiple mRNA species that are induced to similar extents upon disruption of protein folding in the ER. In contrast, HUGT2 is transcribed as a single mRNA species that is not induced under similar conditions. HUGT1 and HUGT2 mRNAs are broadly expressed in multiple tissues and differ slightly in their tissue distribution. The HUGT1 and HUGT2 cDNAs were expressed by transient transfection in COS-1 monkey cells to obtain similar levels of protein localized to the ER. Extracts from HUGT1-transfected cells displayed a 27-fold increase in the transfer of [(14)C]glucose from UDP [(14)C]glucose to denatured substrates. Despite its high degree of sequence identity with HUGT1, the expressed recombinant HUGT2 protein was not functional under the conditions optimized for HUGT1. Site-directed alanine mutagenesis within a highly conserved region of HUGT1 identified four residues that are essential for catalytic function. PMID- 10694381 TI - Role of the loop containing residue 115 in the induced-fit mechanism of the bacterial cell wall biosynthetic enzyme MurA. AB - The induced-fit mechanism in Enterobacter cloacae MurA has been investigated by kinetic studies and X-ray crystallography. The antibiotic fosfomycin, an irreversible inhibitor of MurA, induced a structural change in UDP-N acetylglucosamine (UDPGlcNAc)-liganded enzyme with a time dependence similar to that observed for the inactivation progress. The mechanism of action of fosfomycin on MurA appeared to be of the bimolecular type, the overall rate constants of inactivation and structural change being = 104 M(-1) s(-1) and = 85 M(-1) s(-1), respectively. Fosfomycin as well as the second MurA substrate, phosphoenolpyruvate (PEP), are known to interact with the side chain of Cys115. Like wild-type MurA, the catalytically inactive single-site mutant protein Cys115Ser structurally interacted with UDPGlcNAc in a rapidly reversible reaction. However, in contrast to wild-type enzyme, binding of PEP to mutant protein induced a rate-limited, biphasic structural change. Fosfomycin did not affect the structure of the mutant protein. The crystal structure of unliganded Cys115Ser MurA at 1.9 A resolution revealed that the overall conformation of the loop comprising residues 112-121 is not influenced by the mutation. However, other than Cys115 in wild-type MurA, Ser115 exhibits two distinct side-chain conformations. A detailed view on the loop revealed the existence of an elaborate hydrogen-bonding network mainly supplied by water molecules, presumably stabilizing its conformation in the unliganded state. The comparison between the known crystal structures of MurA, together with the kinetic data obtained, suggest intermediate conformational states in the MurA reaction, in which the loop undergoes multiple structural changes upon ligand binding. PMID- 10694382 TI - Determination of metal ion binding sites within the hairpin ribozyme domains by NMR. AB - Cations play an important role in RNA folding and stabilization. The hairpin ribozyme is a small catalytic RNA consisting of two domains, A and B, which interact in the transition state in an ion-dependent fashion. Here we describe the interaction of mono-, di-, and trivalent cations with the domains of the ribozyme, as studied by homo- and heteronuclear NMR spectroscopy. Paramagnetic line broadening, chemical shift mapping, and intermolecular NOEs indicate that the B domain contains four to five metal binding sites, which bind Mn(2+), Mg(2+), and Co(NH(3))(6)(3+). There is no significant structural change in the B domain upon the addition of Co(NH(3))(6)(3+) or Mg(2+). No specific monovalent ion binding sites exist on the B domain, as determined by (15)NH(4)(+) binding studies. In contrast to the B domain, there are no observable metal ion interactions within the internal loop of the A domain. Model structure calculations of Mn(2+) interactions at two sites within the B domain indicate that the binding sites comprise major groove pockets lined with functional groups oriented so that multiple hydrogen bonds can be formed between the RNA and Mn(H(2)O)(6)(2+) or Co(NH(3))(6)(3+). Site 1 is very similar in geometry to a site within the P4-P6 domain of the Tetrahymena group I intron, while site 2 is unique among known ion binding sites. The site 2 ion interacts with a catalytically essential nucleotide and bridges two phosphates. Due to its location and geometry, this ion may play an important role in the docking of the A and B domains. PMID- 10694383 TI - Critical role of tyrosine 277 in the ligand-binding and transactivating properties of retinoic acid receptor alpha. AB - Retinoic acid receptors specifically bind all-trans-retinoic acid (RA) and function as RA-inducible transcriptional regulatory factors. Binding of RA to RARalpha, beta, and gamma is sensitive to nitration with tetranitromethane, a tyrosine-specific modifying reagent. To identify tyrosine residue(s) that are important for RA binding, we carried out chemical modification experiments with purified RARalpha ligand-binding domain (RARalpha-LBD) subjected to partial acid hydrolysis and selective proteolysis. The chemically modified peptides containing each of the three Tyr residues present in the RARalpha-LBD sequence were then analyzed and identified by high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC/ESI-MS). We found that RA binding to RARalpha-LBD protected Tyr(277)-containing peptides from nitration. Protection of Tyr(277) could result either from direct masking by the bound ligand or from ligand-induced changes in receptor conformation and tyrosine accessibility. The role of Tyr residues was further documented by site directed mutagenesis using three site-specific RARalpha mutants: Y208A, Y277A, and Y362A. The affinity for RA of these mutant receptors was in the range of that of the wild-type protein, except for the Y277A receptor mutant, which displays a 15-20-fold reduction in affinity and transactivation activity for RA. Whereas mutation of Tyr(277) into alanine had a variable effect on different agonists and antagonists binding, it caused a dramatic decrease of retinoid-dependent transactivation activity. This later effect was also observed with mutation of Tyr(277) into phenylalanine. It is unlikely that major conformational changes are responsible for the lower affinity of RA binding and RA-dependent transactivation since these mutants displayed wild-type dimerization and DNA-binding activities. Limited proteolysis revealed that upon ligand binding, the Y277A mutant induced a conformational change slightly different from that obtained with the wild-type protein. These data could suggest that Tyr(277) play a critical role in the ligand-induced conformational changes required for the activation of RARalpha. PMID- 10694384 TI - Evaluation by site-directed mutagenesis of aspartic acid residues in the metal site of pig heart NADP-dependent isocitrate dehydrogenase. AB - Pig heart NADP-dependent isocitrate dehydrogenase requires a divalent metal cation for catalysis. On the basis of affinity cleavage studies [Soundar and Colman (1993) J. Biol. Chem. 268, 5267] and analysis of the crystal structure of E. coli NADP-isocitrate dehydrogenase [Hurley et al. (1991) Biochemistry 30, 8671], the residues Asp(253), Asp(273), Asp(275), and Asp(279) were selected as potential ligands of the divalent metal cation in the pig heart enzyme. Using a megaprimer PCR method, the Asp at each of these positions was mutated to Asn. The wild-type and mutant enzymes were expressed in Escherichia coli and purified. D253N has a specific activity, K(m) values for Mn(2+), isocitrate, and NADP, and also a pH-V(max) profile similar to those of the wild-type enzyme. Thus, Asp(253) is not involved in enzyme function. D273N has an increased K(m) for Mn(2+) and isocitrate with a specific activity 5% that of wild type. The D273N mutation also prevents the oxidative metal cleavage seen with Fe(2+) alone in the wild-type enzyme. As compared to wild type, D275N has greatly increased K(m) values for Mn(2+) and isocitrate, with a specific activity <0.1% that of wild type, and a large increase in pK(a) for the enzyme-substrate complex. D279N has only small increases in K(m) for Mn(2+) and isocitrate, but a specific activity <0.1% that of wild type and a major change in the shape of its pH-V(max) profile. These results suggest that Asp(273) and Asp(275) contribute to metal binding, whereas Asp(279), as well as Asp(275), is critical for catalysis. Asp(279) may function as the catalytic base. Using the Modeler program of Insight II, a structure for porcine NADP-isocitrate dehydrogenase was built based on the X-ray coordinates of the E. coli enzyme, allowing visualization of the metal-isocitrate site. PMID- 10694385 TI - HIV-1 protease inhibitors: enthalpic versus entropic optimization of the binding affinity. AB - Existing experimental as well as computational screening methods select potential ligands or drug candidates on the basis of binding affinity. Since the binding affinity is a function of the enthalpy (DeltaH) and entropy (DeltaS) changes, it is apparent that improved binding can be achieved in different ways: by optimizing DeltaH, DeltaS, or a combination of both. However, the behavior of enthalpically or entropically optimized inhibitors is fundamentally different, including their response to mutations that may elicit drug resistance. In the design of HIV-1 protease inhibitors, high binding affinity has usually been achieved by preshaping lead compounds to the geometry of the binding site and by incorporating a high degree of hydrophobicity. The thermodynamic consequence of that approach is that the binding affinity of the resulting inhibitors becomes entropically favorable but enthalpically unfavorable. Specifically, the resulting high binding affinity is due to an increased solvation entropy (hydrophobic effect) combined with a reduced loss of conformational entropy of the inhibitor upon binding (structural rigidity). Here we report that tripeptide inhibitors derived from the transframe region of Gag-Pol (Glu-Asp-Leu and Glu-Asp-Phe) bind to the HIV-1 protease with a favorable enthalpy change. This behavior is qualitatively different from that of known inhibitors and points to new strategies for inhibitor design. Since the binding affinities of enthalpically favorable and enthalpically unfavorable inhibitors have opposite temperature dependence, it is possible to design fast screening protocols that simultaneously select inhibitors on the basis of affinity and enthalpy. PMID- 10694386 TI - Crystal structure and site-specific mutagenesis of pterin-bound human phenylalanine hydroxylase. AB - The crystal structure of the dimeric catalytic domain (residues 118-424) of human PheOH (hPheOH), cocrystallized with the oxidized form of the cofactor (7,8 dihydro-L-biopterin, BH(2)), has been determined at 2.0 A resolution. The pterin binds in the second coordination sphere of the catalytic iron (the C4a atom is 6.1 A away), and interacts through several hydrogen bonds to two water molecules coordinated to the iron, as well as to the main chain carbonyl oxygens of Ala322, Gly247, and Leu249 and the main chain amide of Leu249. Some important conformational changes are seen in the active site upon pterin binding. The loop between residues 245 and 250 moves in the direction of the iron, and thus allows for several important hydrogen bonds to the pterin ring to be formed. The pterin cofactor is in an ideal orientation for dioxygen to bind in a bridging position between the iron and the pterin. The pterin ring forms an aromatic pi-stacking interaction with Phe254, and Tyr325 contributes to the positioning of the pterin ring and its dihydroxypropyl side chain by hydrophobic interactions. Of particular interest in the hPheOH x BH(2) binary complex structure is the finding that Glu286 hydrogen bonds to one of the water molecules coordinated to the iron as well as to a water molecule which hydrogen bonds to N3 of the pterin ring. Site-specific mutations of Glu286 (E286A and E286Q), Phe254 (F254A and F254L), and Tyr325 (Y325F) have confirmed the important contribution of Glu286 and Phe254 to the normal positioning of the pterin cofactor and catalytic activity of hPheOH. Tyr325 also contributes to the correct positioning of the pterin, but has no direct function in the catalytic reaction, in agreement with the results obtained with rat TyrOH [Daubner, S. C., and Fitzpatrick, P. F. (1998) Biochemistry 37, 16440-16444]. Superposition of the binary hPheOH.BH(2) complex onto the crystal structure of the ligand-free rat PheOH (which contains the regulatory and catalytic domains) [Kobe, B., Jennings, I. G., House, C. M., Michell, B. J., Goodwill, K. E., Santarsiero, B. D., Stevens, R. C., Cotton, R. G. H., and Kemp, B. E. (1999) Nat. Struct. Biol. 6, 442-448] reveals that the C2' hydroxyl group of BH(2) is sufficiently close to form hydrogen bonds to Ser23 in the regulatory domain. Similar interactions are seen with the hPheOH.adrenaline complex and Ser23. These interactions suggest a structural explanation for the specific regulatory properties of the dihydroxypropyl side chain of BH(4) (negative effector) in the full-length enzyme in terms of phosphorylation of Ser16 and activation by L-Phe. PMID- 10694387 TI - Escherichia coli methionyl-tRNA formyltransferase: role of amino acids conserved in the linker region and in the C-terminal domain on the specific recognition of the initiator tRNA. AB - The formylation of initiator methionyl-tRNA by methionyl-tRNA formyltransferase (MTF) is important for the initiation of protein synthesis in eubacteria. We are studying the molecular mechanisms of recognition of the initiator tRNA by Escherichia coli MTF. MTF from eubacteria contains an approximately 100-amino acid C-terminal extension that is not found in the E. coli glycinamide ribonucleotide formyltransferase, which, like MTF, use N(10) formyltetrahydrofolate as a formyl group donor. This C-terminal extension, which forms a distinct structural domain, is attached to the N-terminal domain through a linker region. Here, we describe the effect of (i) substitution mutations on some nineteen basic, aromatic and other conserved amino acids in the linker region and in the C-terminal domain of MTF and (ii) deletion mutations from the C terminus on enzyme activity. We show that the positive charge on two of the lysine residues in the linker region leading to the C-terminal domain are important for enzyme activity. Mutation of some of the basic amino acids in the C terminal domain to alanine has mostly small effects on the kinetic parameters, whereas mutation to glutamic acid has large effects. However, the deletion of 18, 20, or 80 amino acids from the C-terminus has very large effects on enzyme activity. Overall, our results support the notion that the basic amino acid residues in the C-terminal domain provide a positively charged channel that is used for the nonspecific binding of tRNA, whereas some of the amino acids in the linker region play an important role in activity of MTF. PMID- 10694388 TI - NBD-isocolcemid-tubulin interaction: a novel one-step reaction involving no conformational adjustment of reactants. AB - Isocolcemid, a colcemid analogue in which the positions of the C-ring methoxy and carbonyl are exchanged, is virtually inactive in binding to tubulin and inhibiting the formation of microtubule assembly. We have found that the substitution of a NBD group in the side chain of the B-ring of isocolcemid can reverse the effect of these structural alterations (at the C-ring) and the newly synthesized NBD-isocolcemid restores the lost biological activity. It inhibits microtubule assembly with an IC(50) of 12 microM and competes efficiently with [(3)H]colchicine, for binding to tubulin. NBD-isocolcemid has two binding sites on tubulin; one is characterized by fast binding, whereas the binding to the other site is slow. These two sites are independent and unrelated to each other. Colchicine and its analogues compete with NBD-isocolcemid for the slow site. Association and dissociation rate constants for the fast site, obtained from the stopped-flow measurements, are (7.37 +/- 0. 70) x 10(5) M(-1) s(-1) and 7.82 +/- 2.74 s(-1), respectively. While the interaction of colchicine and its analogues with tubulin involves two steps, NBD-isocolcemid binding to tubulin at the slow site has been found to be a one-step reaction. This is evident from the linear dependence of the observed rate constant (k(obs)) with both NBD-isocolcemid and tubulin concentrations. The interaction of NBD-isocolcemid with tubulin does not involve the conformational change of NBD-isocolcemid, as is evident from the unchanged CD spectra of the drug. The absence of enhanced GTPase activity of tubulin and the native-like protease cleavage pattern of the NBD-isocolcemid tubulin complex suggest an unaltered conformation of tubulin upon NBD-isocolcemid binding to it as well. Implications of this on the mechanism of polymerization inhibition have been discussed. PMID- 10694389 TI - Neighboring aliphatic/aromatic side chain interactions between residues 9 and 10 in gramicidin channels. AB - The interactions between an aliphatic or phenyl side chain and an indole ring in a phospholipid environment were investigated by synthesizing and characterizing gramicidins in which Trp(9) was ring-labeled and D-Leu(10) was replaced by D-Val, D-Ala, or D-Phe. All three analogues form conducting channels, with conductances that are lower than that of gramicidin A (gA) channels. The channel lifetimes vary by less than 50% from that of gA channels. Circular dichroism spectra and size-exclusion chromatography show that the conformation of each analogue in dimyristoylphosphatidylcholine (DMPC) vesicles is similar to the right-handed beta(6.3)-helical conformation that is observed for gA. (2)H NMR spectra of oriented samples in DMPC show large changes for the Trp(9) ring when residue 10 is modified, suggesting a steric interaction between D-Leu(10) and Trp(9), in agreement with previous acylation studies (R. E. Koeppe II et al. (1995) Biochemistry 34, 9299-9307). The outer quadrupolar splitting for Trp(9) is unchanged with D-Phe(10), at approximately 153 kHz, but increases by approximately 25 kHz with D-Val(10) and decreases by approximately 10 kHz with D Ala(10). With D-Ala(10) or D-Val(10), the outer resonance splits into two in a temperature-dependent manner. The NMR spectra indicate that the side chain torsion angles chi1 and chi2 for Trp(9) change when residue 10 is substituted. The changes in chi1 are small, in all cases less than 10 degrees, as is Deltachi2 when D-Ala(10) is introduced, but with D-Val(10) and D-Phe(10) Deltachi2 is at least 25 degrees. We conclude that D-Leu(10) helps to stabilize an optimal orientation of Trp(9) in gA channels in lipid bilayers and that changes in Trp orientation alter channel conductance and lifetime without affecting the basic channel fold. PMID- 10694390 TI - Recognition of Dictyostelium discoideum lysosomal enzymes is conferred by the amino-terminal carbohydrate binding site of the insulin-like growth factor II/mannose 6-phosphate receptor. AB - The insulin-like growth factor-II/mannose 6-phosphate receptor (IGF-II/MPR) is a type I glycoprotein that mediates both the intracellular sorting of lysosomal enzymes bearing mannose 6-phosphate (Man-6-P) residues to the lysosome and the bioavailability of IGF-II. The extracytoplasmic region of the IGF-II/MPR contains 15 repeating domains; the two carbohydrate recognition domains (CRDs) have been localized to domains 1-3 and 7-9, and the high-affinity IGF-II binding site maps to domain 11. To characterize the carbohydrate binding properties of the IGF II/MPR, regions of the receptor encompassing the individual CRDs were produced in a baculovirus expression system. Characterization of the recombinant proteins revealed that the pH optimum for carbohydrate binding is significantly more acidic for the carboxyl-terminal CRD than for the amino-terminal CRD (i.e., pH 6.4-6.5 vs 6.9). Equilibrium binding studies demonstrated that the two CRDs exhibit a similar affinity for Man-6-P. Furthermore, substitution of the conserved arginine residue in domain 3 (R435) or in domain 9 (R1334) with alanine resulted in a similar >1000-fold decrease in the affinity for the lysosomal enzyme, beta-glucuronidase. In contrast, the two CRDs differ dramatically in their ability to recognize the distinctive modifications (i.e., mannose 6-sulfate and Man-6-P methyl ester) found on Dictyostelium discoideum lysosomal enzymes: the amino-terminal CRD binds mannose 6-sulfate and Man-6-P methyl ester with a 14 55-fold higher affinity than the carboxyl-terminal CRD. Taken together, these results demonstrate that the IGF-II/MPR contains two functionally distinct CRDs. PMID- 10694391 TI - The interaction between the regulatory light chain domains on two heads is critical for regulation of smooth muscle myosin. AB - Recent findings have suggested that the interaction between the two heads is critical for phosphorylation-dependent regulation of smooth muscle myosin. We hypothesized that the interaction between the two regulatory light chains on two heads of myosin dictates the regulation of myosin motor function. To evaluate this notion, we engineered and characterized smooth muscle heavy meromyosin (HMM), which is composed of one entire HMM heavy chain and one motor domain truncated heavy chain containing the S2 rod and regulatory light chain (RLC) binding site, as well as the bound RLC (SMDHMM). SMDHMM was inactive for both actin-translocating activity and actin-activated ATPase activity in the dephosphorylated state, demonstrating that the interaction between the two RLC domains on the two heads and/or a motor domain and a RLC domain in a distinct head is sufficient for the inhibition of smooth muscle myosin motor activity. When phosphorylated, SMDHMM was activated for both actin-translocating activity and actin-activated ATPase activity; however, these activities were lower than those of double-headed HMM, implying partial release of inhibition by phosphorylation in SMDHMM and/or cooperativity between the two heads of smooth muscle myosin. The present results indicate that the RLC domain is critical for phosphorylation-dependent regulation of smooth muscle myosin motor activity. On the other hand, similar to double-headed HMM, SMDHMM showed both "folded" and "extended" conformations, and the ratio of those conformations is dependent on ionic strength, suggesting that the RLC domain is sufficient to regulate the conformational transition in myosin. PMID- 10694392 TI - Use of an altered sugar-nucleotide to unmask the transition state for alpha(2- >6) sialyltransferase. AB - Rat liver alpha(2-->6) sialyltransferase catalyzes the formation of a glycosidic bond between N-acetylneuraminic acid and the 6-hydroxyl group of a galactose residue at the nonreducing terminus of an oligosaccharide. This reaction has been investigated through the use of the novel sugar-nucleotide donor substrate UMP NeuAc. A series of UMP-NeuAc radioisotopomers were prepared by chemical deamination of the corresponding CMP-NeuAc precursors. Kinetic isotope effects (KIEs) on V/K were measured using mixtures of radiolabeled UMP-NeuAc's as the donor substrate and N-acetyllactosamine as the acceptor. The secondary beta-(2)H KIE was 1.218 +/- 0.010, and the primary (14)C KIE was 1.030 +/- 0.010. A large inverse (3)H binding isotope effect of 0.944 +/- 0.010 was measured at the terminal carbon of the NeuAc glycerol side chain. These KIEs observed using UMP NeuAc are much larger than those previously measured with CMP-NeuAc [Bruner, M., and Horenstein, B. A. (1998) Biochemistry 37, 289-297]. Solvent deuterium isotope effects of 1.3 and 2.6 on V/K and V(max) were observed with CMP-NeuAc as the donor, and it is revealing that these isotope effects vanished with use of the slow donor substrate UMP-NeuAc. Bell-shaped pH versus rate profiles were observed for V(max) (pK(a) values = 5.5, 9.0) and V/K(UMP)(-)(NeuAc) (pK(a)values = 6.2, 9.0). The results are considered in terms of a mechanism involving an isotopically sensitive conformational change which is independent of the glycosyl transfer step. The isotope effects reveal that the enzyme-bound transition state bears considerable charge on the N-acetylneuraminic acid residue, and this and other features of this mechanism provide new directions for sialyltransferase inhibitor design. PMID- 10694393 TI - Engineering of Sulfolobus solfataricus HMG-CoA reductase to a form whose activity is regulated by phosphorylation and dephosphorylation. AB - There are two classes of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase: the class I enzymes of eukaryotes and some archaea, and the class II enzymes of certain eubacteria. The activity of the class I Syrian hamster HMG-CoA reductase is regulated by phosphorylation-dephosphorylation of Ser871. Phosphorylation apparently prevents the active site histidine, His865, from protonating the inhibitory coenzyme A thioanion prior to its release from the enzyme. Structural evidence for this hypothesis is, however, lacking. The HMG-CoA reductase of the thermophilic archaeon Sulfolobus solfataricus, whose stability recommends it for physical studies, lacks both a phosphoacceptor serine and a protein kinase recognition motif. Consequently, its activity is not regulated by phosphorylation. We therefore employed site-directed mutagenesis to engineer an appropriately located phosphoacceptor serine and cAMP-dependent protein kinase recognition motif. Substitution of serine for Ala406, the apparent cognate of hamster Ser871, and replacement of Leu403 and Gly404 by arginine created S. solfataricus mutant enzyme L403R/G404R/A406S. The general properties of enzyme L403R/G404R/A406S (K(m) values, V(max), optimal pH and temperature) were essentially those of the wild-type enzyme. Exposure of enzyme L403R/G404R/A406S to [gamma-(32)P]ATP and cAMP-dependent protein kinase was accompanied by incorporation of (32)P(i) and by a parallel decrease in catalytic activity. Subsequent treatment with a protein phosphatase released enzyme-bound (32)P(i) and restored activity to pretreatment levels. The regulatory properties of enzyme L403R/G404R/A406S thus match those of the hamster enzyme. Solution of the three dimensional structures of the phospho and dephospho forms of this mutant enzyme thus should reveal structural features critical for regulation of the activity of a class I HMG-CoA reductase. PMID- 10694394 TI - Stereochemistry of the enolization of scytalone by scytalone dehydratase. AB - In D(2)O, scytalone exchanges its two C2 hydrogen atoms for deuterium atoms at different rates. At pD 7.0 and 25 degrees C, half-lives for the exchanges are 0.8 and 10 days for the pro-S and pro-R hydrogens, respectively. The differential exchange rates allow for the preparation of multiple scytalone samples (through incubation of scytalone in D(2)O and then back exchanging with H(2)O) having differential levels of deuterium enrichment at the C2 pro-S and pro-R positions. From these samples, the stereochemical preference for hydrogen abstraction during the dehydration reaction mediated by the enzyme scytalone dehydratase was determined. At pH 7. 0, deuterium at the pro-S position has little effect on enzyme catalysis, whereas deuterium at the pro-R position produces kinetic isotope effects of 2.3 (25 degrees C), 5.1 (25 degrees C), and 6.7 (6.8 degrees C) on k(cat), k(cat)/K(m), and the single-turnover rate, respectively. The results are fully consistent with the enzyme catalyzing a syn elimination through an E1cb-like mechanism. The syn elimination is compatible with the interactions realized between a scytalone boat conformation and key active site residues as modeled from multiple X-ray crystal structures of the enzyme in complexes with inhibitors. PMID- 10694395 TI - Kinetics and mechanism of the citrate synthase from the thermophilic archaeon Thermoplasma acidophilum. AB - The kinetics and mechanism of the citrate synthase from a moderate thermophile, Thermoplasma acidophilum (TpCS), are compared with those of the citrate synthase from a mesophile, pig heart (PCS). All discrete steps in the mechanistic sequence of PCS can be identified in TpCS. The catalytic strategies identified in PCS, destabilization of the oxaloacetate substrate carbonyl and stabilization of the reactive species, acetyl-CoA enolate, are present in TpCS. Conformational changes, which allow the enzyme to efficiently catalyze both condensation of acetyl-CoA thioester and subsequently hydrolysis of citryl-CoA thioester within the same active site, occur in both enzymes. However, significant differences exist between the two enzymes. PCS is a characteristically efficient enzyme: no internal step is clearly rate-limiting and the condensation step is readily reversible. TpCS is a less efficient catalyst. Over a broad temperature range, inadequate stabilization of the transition state for citryl-CoA hydrolysis renders this step nearly rate-limiting for the forward reaction of TpCS. Further, excessive stabilization of the citryl-CoA intermediate renders the condensation step nearly irreversible. Values of substrate and solvent deuterium isotope effects are consistent with the kinetic model. Near its temperature optimum (70 degrees C), there is a modest increase in the reversibility of the condensation step for TpCS, but reversibility still falls short of that shown by PCS at 37 degrees C. The root cause of the catalytic inefficiency of TpCS may lie in the lack of protein flexibility imposed by the requirement for thermal stability of the protein itself or its temperature-labile substrate, oxaloacetate. PMID- 10694396 TI - Selectivity of the yersiniabactin synthetase adenylation domain in the two-step process of amino acid activation and transfer to a holo-carrier protein domain. AB - The adenylation (A) domain of the Yersinia pestis nonribosomal peptide synthetase that biosynthesizes the siderophore yersiniabactin (Ybt) activates three molecules of L-cysteine and covalently aminoacylates the phosphopantetheinyl (P pant) thiols on three peptidyl carrier protein (PCP) domains embedded in the two synthetase subunits, two in cis (PCP1, PCP2) in subunit HMWP2 and one in trans (PCP3) in subunit HMWP1. This two-step process of activation and loading by the A domain is analogous to the operation of the aminoacyl-tRNA synthetases in ribosomal peptide synthesis. Adenylation domain specificity for the first step of reversible aminoacyl adenylate formation was assessed with the amino acid dependent [(32)P]-PP(i)-ATP exchange assay to show that S-2-aminobutyrate and beta-chloro-L-alanine were alternate substrates. The second step of A domain catalysis, capture of the bound aminoacyl adenylate by the P-pant-SH of the PCP domains, was assayed both by catalytic release of PP(i) and by covalent aminoacylation of radiolabeled substrates on either the PCP1 fragment of HMWP2 or the PCP3-thioesterase double domain fragment of HMWP1. There was little selectivity for capture of each of the three adenylates by PCP3 in the second step, arguing against any hydrolytic proofreading of incorrect substrates by the A domain. The holo-PCP3 domain accelerated PP(i) release and catalytic turnover by 100-200-fold over the leak rate (<1 min(-1)) of aminoacyl adenylates into solution while PCP1 in trans had only about a 5-fold effect. Free pantetheine could capture cysteinyl adenylate with a 25-50-fold increase in k(cat) while CoA was 10-fold less effective. The K(m) of free pantetheine (30-50 mM) was 3 orders of magnitude larger than that of PCP3-TE (10-25 microM), indicating a net 10(4) greater catalytic efficiency for transfer to the P-pant arm of PCP3 by the Ybt synthetase A domain, relative to P-pant alone. PMID- 10694397 TI - A molecular coupling mechanism for the oxaloacetate decarboxylase Na+ pump as inferred from mutational analysis. AB - The oxaloacetate decarboxylase Na+ pump consists of subunits alpha, beta, and gamma, and contains biotin as the prosthetic group. Membrane-bound subunit beta catalyzes the decarboxylation of carboxybiotin coupled to Na+ translocation, and consumes a periplasmically derived proton. Site-directed mutagenesis of conserved amino acids of transmembrane helix VIII indicated that residues N373, G377, S382, and R389 are functionally important. The polar side groups of these amino acids may constitute together with D203 a network of ionizable groups which promotes the translocation of Na+ and the oppositely oriented H+ across the membrane. Evidence is presented that two Na+ ions are bound simultaneously to subunit beta during transport with D203 and S382 acting as binding sites. Sodium ion binding from the cytoplasm to both sites elicits decarboxylation of carboxybiotin, and a conformational switch exposes the bound Na+ ions toward the periplasm. After dissociation of Na+ and binding of H+, the cytoplasmically exposed conformation is regained. PMID- 10694398 TI - Spectroscopic analysis of the trinuclear cluster in the Fet3 protein from yeast, a multinuclear copper oxidase. AB - The Fet3 protein (Fet3p) is a multinuclear copper oxidase essential for high affinity iron uptake in yeast. Fet3p contains one type 1, one type 2, and a strongly antiferromagnetically coupled binuclear Cu(II)-Cu(II) type 3 copper. The type 2 and type 3 sites constitute a structurally distinct trinuclear cluster at which dioxygen is reduced to water. In Fet3p, as in ceruloplasmin, Fe(II) is oxidized to Fe(III) at the type 1 copper; this is the ferroxidase reaction that is fundamental to the physiologic function of these two enzymes. Using site directed mutagenesis, we have generated type 1-depleted (T1D), type 2-depleted (T2D), and T1D/T2D mutants. None were active in the essential ferroxidase reaction catalyzed by Fet3p. However, the spectroscopic signatures of the remaining Cu(II) sites in any one of the three mutants were indistinguishable from those exhibited by the wild type. Although the native protein and the T1D mutant were isolated in the completely oxidized Cu(II) form, the T2D and T1D/T2D mutants were found to be completely reduced. This result is consistent with the essential role of the type 2 copper in dioxygen turnover, and with the suggestions that cuprous ion is the valence state of intracellular copper. Although stable to dioxygen, the Cu(I) sites in both proteins were readily oxidized by hydrogen peroxide. The double mutant was extensively analyzed by X ray absorption spectroscopy. Edge and near-edge features clearly distinguished the oxidized from the reduced form of the binuclear cluster. EXAFS was strongly consistent with the expected coordination of each type 3 copper by three histidine imidazoles. Also, copper scattering was observed in the oxidized cluster along with scattering from a ligand corresponding to a bridging oxygen. The data derived from the reduced cluster indicated that the bridge was absent in this redox state. In the reduced form of the double mutant, an N/O ligand was apparent that was not seen in the reduced form of the T1D protein. This ligand in T1D/T2D could be either the remaining type 2 copper imidazole ligand (from His416) or a water molecule that could be stabilized at the type 3 cluster by H bonding to this side chain. If present in the native protein, this H(2)O could provide acid catalysis of dioxygen reduction at the reduced trinuclear center. PMID- 10694399 TI - Evidence for the rate of the final step in the bacteriorhodopsin photocycle being controlled by the proton release group: R134H mutant. AB - Light absorbed by bacteriorhodopsin (bR) leads to a proton being released at the extracellular surface of the purple membrane. Structural studies as well as studies of mutants of bR indicate that several groups form a pathway for proton transfer from the Schiff base to the extracellular surface. These groups include D85, R82, E204, E194, and water molecules. Other residues may be important in tuning the initial state pK(a) values of these groups and in mediating light induced changes of the pK(a) values. A potentially important residue is R134: it is located close to E194 and might interact electrostatically to affect the pK(a) of E194 and light-induced proton release. In this study we investigated effects of the substitution of R134 with a histidine on light-induced proton release and on the photocycle transitions associated with proton transfer. By measuring the light-induced absorption changes versus pH, we found that the R134H mutation results in an increase in the pK(a) of the proton release group in both the M (0.6 pK unit) and O (0.7 pK unit) intermediate states. This indicates the importance of R134 in tuning the pK(a) of the group that, at neutral and high pH, releases the proton upon M formation (fast proton release) and that, at low pH, releases the proton simultaneously with O decay (slow proton release). The higher pK(a) of the proton release group found in R134H correlates with the slowing of the rate of the O --> bR transition at low pH and probably is the cause of this slowing. The pH dependence of the fraction of the O intermediate is altered in R134H compared to the WT but is similar to that in the E194D mutant: a very small amount of O is present at neutral pH, but the fraction of O increases greatly upon decreasing the pH. These results provide further support for the hypothesis that the O --> bR transition is controlled by the rate of deprotonation of the proton release group. These data also provide further evidence for the importance of the R134-E194 interaction in modulating proton release from D85 after light has led to its being protonated. PMID- 10694400 TI - Formation and reactions of the heme-dioxygen intermediate in the first and second steps of nitric oxide synthesis as studied by stopped-flow spectroscopy under single-turnover conditions. AB - To better understand the mechanism of nitric oxide (NO) synthesis, we studied conversion of N-hydroxy-L-arginine (NOHA) or L-arginine (Arg) to citrulline and NO under single-turnover conditions using the oxygenase domain of neuronal nitric oxide synthase (nNOSoxy) and rapid scanning stopped-flow spectroscopy. When anaerobic nNOSoxy saturated with H(4)B and NOHA was provided with 0.5 or 1 electron per heme and then exposed to air at 25 degrees C, it formed 0.5 or 1 mol of citrulline/mol of heme, respectively, indicating that NOHA conversion had 1:1 stoichiometry with respect to electrons added. Identical experiments with Arg produced substoichiometric amounts of NOHA or citrulline even when up to 3 electrons were provided per heme. Transient spectral intermediates were investigated at 10 degrees C. For NOHA, four species were observed in the following sequence: starting ferrous nNOSoxy, a transient ferrous-dioxygen complex, a transient ferric-NO complex, and ferric nNOSoxy. For Arg, transient intermediates other than the ferrous-dioxygen species were not apparent during the reaction. Our results provide a kinetic framework for formation and reactions of the ferrous-dioxygen complex in each step of NO synthesis and establish that (1) the ferrous-dioxy enzyme reacts quantitatively with NOHA but not with Arg and (2) its reaction with NOHA forms 1 NO/heme, which immediately binds to form a ferric heme-NO complex. PMID- 10694401 TI - Demonstration of a conserved histidine and two water ligands at the Mn2+ site in Diocleinae lectins by pulsed EPR spectroscopy. AB - Lectins from the Diocleinae subtribe, including Canavalia brasiliensis, Canavalia bonariensis, Canavalia grandiflora, Cratylia floribunda, Dioclea grandiflora, Dioclea guianensis, Dioclea rostrata, Dioclea violacea, and Dioclea virgata, have been recently isolated and characterized in terms of their carbohydrate binding specificities. Although all of the lectins are Man/Glc specific, they possess different biological activities. In the present study, electron paramagnetic resonance (EPR) spectroscopy demonstrates that all nine Diocleinae lectins contain Mn2+. The spectra of C. floribunda and D. rostrata suggest Mn2+ site symmetry different from that of the other seven lectins. However, electron spin echo envelope modulation (ESEEM) spectroscopy indicates that all nine lectins are coordinated to a histidyl imidazole, with similar electron-nuclear coupling to the Mn2+-bound imidazole nitrogen. ESEEM also demonstrates ligation of two water molecules to Mn2+ in all nine Diocleinae lectins. Thus, the EPR and ESEEM data indicate the presence of a Mn2+ binding site in the above Diocleinae lectins with a conserved histidine residue and two water ligands. PMID- 10694403 TI - Nonlinear electron paramagnetic resonance studies of the interaction of cytochrome c oxidase with spin-labeled lipids in gel-phase membranes. AB - The interaction of lipids, spin-labeled at different positions in the sn-2 chain, with cytochrome c oxidase reconstituted in gel-phase membranes of dimyristoylphosphatidylglycerol has been studied by electron paramagnetic resonance (EPR) spectroscopy. Nonlinear EPR methods, both saturation transfer EPR and progressive saturation EPR, were used. Interaction with the protein largely removes the flexibility gradient of the lipid chains in gel-phase membranes. The rotational mobility of the chain segments is reduced, relative to that for gel phase lipids, by the intramembranous interaction with cytochrome c oxidase. This holds for all positions of chain labeling, but the relative effect is greater for chain segments closer to the terminal methyl ends. Modification of the paramagnetic metal-ion centers in the protein by binding azide has a pronounced effect on the spin-lattice relaxation of the lipid spin labels. This demonstrates that the centers modified are sufficiently close to the first-shell lipids to give appreciable dipolar interactions and that their vertical location in the membrane is closer to the 5-position than to the 14-position of the lipid chains. PMID- 10694402 TI - Metal requirements of a diadenosine pyrophosphatase from Bartonella bacilliformis: magnetic resonance and kinetic studies of the role of Mn2+. AB - Recombinant IalA protein from Bartonella bacilliformis is a monomeric adenosine 5'-tetraphospho-5'-adenosine (Ap4A) pyrophosphatase of 170 amino acids that catalyzes the hydrolysis of Ap4A, Ap5A, and Ap6A by attack at the delta phosphorus, with the departure of ATP as the leaving group [Cartwright et al. (1999) Biochem. Biophys. Res. Commun. 256, 474-479]. When various divalent cations were tested over a 300-fold concentration range, Mg2+, Mn2+, and Zn2+ ions were found to activate the enzyme, while Ca2+ did not. Sigmoidal activation curves were observed with Mn2+ and Mg2+ with Hill coefficients of 3.0 and 1.6 and K0.5 values of 0.9 and 5.3 mM, respectively. The substrate M2+ x Ap4A showed hyperbolic kinetics with Km values of 0.34 mM for both Mn2+ x Ap4A and Mg2+ x Ap4A. Direct Mn2+ binding studies by electron paramagnetic resonance (EPR) and by the enhancement of the longitudinal relaxation rate of water protons revealed two Mn2+ binding sites per molecule of Ap4A pyrophosphatase with dissociation constants of 1.1 mM, comparable to the kinetically determined K0.5 value of Mn2+. The enhancement factor of the longitudinal relaxation rate of water protons due to bound Mn2+ (epsilon b) decreased with increasing site occupancy from a value of 12.9 with one site occupied to 3.3 when both are occupied, indicating site site interaction between the two enzyme-bound Mn2+ ions. Assuming the decrease in epsilon(b) to result from cross-relaxation between the two bound Mn2+ ions yields an estimated distance of 5.9 +/- 0.4 A between them. The substrate Ap4A binds one Mn2+ (Kd = 0.43 mM) with an epsilon b value of 2.6, consistent with the molecular weight of the Mn2+ x Ap4A complex. Mg2+ binding studies, in competition with Mn2+, reveal two Mg2+ binding sites on the enzyme with Kd values of 8.6 mM and one Mg2+ binding site on Ap4A with a Kd of 3.9 mM, values that are comparable to the K0.5 for Mg2+. Hence, with both Mn2+ and Mg2+, a total of three metal binding sites were found-two on the enzyme and one on the substrate-with dissociation constants comparable to the kinetically determined K0.5 values, suggesting a role in catalysis for three bound divalent cations. Ca2+ does not activate Ap4A pyrophosphatase but inhibits the Mn2+-activated enzyme competitively with a Ki = 1.9 +/- 1.3 mM. Ca2+ binding studies, in competition with Mn2+, revealed two sites on the enzyme with dissociation constants (4.3 +/- 1.3 mM) and one on Ap4A with a dissociation constant of 2.1 mM. These values are similar to its Ki suggesting that inhibition by Ca2+ results from the complete displacement of Mn2+ from the active site. Unlike the homologous MutT pyrophosphohydrolase, which requires only one enzyme-bound divalent cation in an E x M2+ x NTP x M2+ complex for catalytic activity, Ap4A pyrophosphatase requires two enzyme-bound divalent cations that function in an active E x (M2+)2 x Ap4A x M2+ complex. PMID- 10694404 TI - Unfolding thermodynamics of the tetrameric chaperone, SecB. AB - SecB is a cytosolic tetrameric chaperone in Escherichia coli, which maintains polypeptides, destined for export in a translocation competent state. The thermodynamics of unfolding of SecB was studied as a function of protein concentration, by using high sensitivity-differential scanning calorimetry and spectroscopic methods. The thermal unfolding of tetrameric SecB is reversible and can be well described as a two-state transition in which the folded tetramer is converted directly to unfolded monomers. Increasing the pH decreases the stability of the tetramer significantly, the T(m) changing from 341.3 K at pH 6.5 to 332.6 K at pH 9.5. The value of DeltaC(p) obtained from measurements of DeltaH(m) as a function of T(m) was 10.7 +/- 0.7 kcal mol(-1) K(-1). The value of DeltaC(p) is among the highest measured for a multimeric protein. At 298 K, pH 7.4, the DeltaG degrees (u) for the SecB tetramer is 27.9 +/- 2 kcal mol(-1). Denaturant-mediated unfolding of SecB was found to be irreversible. The reactivity of the four solvent-exposed free thiols in tetrameric SecB is salt dependent. The kinetics of reactivity suggests that these four cysteines are in close proximity to each other and that these residues on each monomer are in chemically identical environments. The thermodynamic data suggest that SecB is a stable, well-folded, and tightly packed tetramer and that substrate binding occurs at a surface site rather than at an interior cavity. PMID- 10694405 TI - Predimerization of recombinant platelet-derived growth factor receptor extracellular domains increases antagonistic potency. AB - Platelet-derived growth factor (PDGF) is a dimeric growth factor acting through tyrosine kinase alpha- and beta-receptors. In both receptors, the extracellular parts are composed of five Ig-like domains. Functional mapping of the extracellular part of the receptors have shown that ligand-binding occurs to Ig like domains 2 and 3 and that Ig-like domain 4 is involved in receptor-receptor interactions. Recombinant GST-fusion proteins of PDGF alpha-receptor Ig-like domains 1-4 and beta-receptor Ig-like domains 1-3 (alphaRD1-4-GST and betaRD1-3 GST) were generated and compared with their cleaved counterparts (alphaRD1-4 and betaRD1-3) with regard to their ability to block PDGF binding to cell surface receptors. In the case of both the alpha- and the beta-receptors, 100-1000-fold lower concentrations of the GST-fusion proteins were required, as compared to the cleaved forms, for inhibition of PDGF binding to cell surface receptors. alphaRD1 4-GST and betaRD1-3-GST, in contrast to alphaRD1-4 and betaRD1-3, were shown to occur as ligand independent dimers. Covalently cross-linked alphaRD1-4 dimers displayed a 50-fold increased potency as compared to alphaRD1-4. We thus conclude that the dimeric nature of alphaRD1-4-GST and betaRD1-3-GST is responsible for the high antagonistic potency of the fusion proteins. PMID- 10694406 TI - Formation of novel D-ring and E-ring isoprostane-like compounds (D4/E4 neuroprostanes) in vivo from docosahexaenoic acid. AB - Free radical-mediated oxidant injury and lipid peroxidation have been implicated in a number of neural disorders. We have reported that bioactive prostaglandin D2/E2-like compounds, termed D2/E2-isoprostanes, are produced in vivo by the free radical-catalyzed peroxidation of arachidonic acid. Docosahexaenoic acid, in contrast to arachidonic acid, is the most abundant unsaturated fatty acid in brain. We therefore questioned whether D/E-isoprostane-like compounds (D4/E4 neuroprostanes) are formed from the oxidation of docosahexaenoic acid. Levels of putative D4/E4-neuroprostanes increased 380-fold after oxidation of docosahexaenoic acid in vitro from 15.2 +/- 6.3 to 5773 +/- 1024 ng/mg of docosahexaenoic acid. Subsequently, chemical approaches and liquid chromatography electrospray ionization tandem mass spectrometry definitively identified these compounds as D4/E4-neuroprostanes. We then explored the formation of D4/E4 neuroprostanes from a biological source, rat brain synaptosomes. Basal levels of D4/E4-neuroprostanes were 3.8 +/- 0.6 ng/mg of protein and increased 54-fold after oxidation (n = 4). We also detected these compounds in fresh brain tissue from rats at levels of 12.1 +/- 2.4 ng/g of brain tissue (n = 3) and in human brain tissue at levels of 9.2 +/- 4.1 ng/g of brain tissue (n = 4). Thus, these studies have identified novel D/E-ring isoprostane-like compounds that are derived from docosahexaenoic acid and that are formed in brain in vivo. The fact that they are readily detectable suggests that ongoing oxidative stress is present in the central nervous system of humans and animals. Further, identification of these compounds provides a rationale for examining their role in neurological disorders associated with oxidant stress. PMID- 10694407 TI - Design of P1' and P3' residues of trivalent thrombin inhibitors and their crystal structures. AB - Synthetic bivalent thrombin inhibitors comprise an active site blocking segment, a fibrinogen recognition exosite blocking segment, and a linker connecting these segments. Possible nonpolar interactions of the P1' and P3' residues of the linker with thrombin S1' and S3' subsites, respectively, were identified using the "Methyl Scan" method [Slon-Usakiewicz et al. (1997) Biochemistry 36, 13494 13502]. A series of inhibitors (4-tert-butylbenzenesulfonyl)-Arg-(D-pipecolic acid)-Xaa-Gly-Yaa-Gly-betaAla-Asp-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala- (be ta cyclohexylalanine)-(D-Glu)-OH, in which nonpolar P1' residue Xaa or P3' residue Yaa was incorporated, were designed and improved the affinity to thrombin. Substitution of the P3' residue with D-phenylglycine or D-Phe improved the K(i) value to (9.5 +/- 0.6) x 10(-14) or 1.3 +/- 0.5 x 10(-13) M, respectively, compared to that of a reference inhibitor with Gly residues at Xaa and Yaa residues (K(i) = (2.4 +/- 0.5) x 10(-11) M). Similarly, substitution of the P1' residue with L-norleucine or L-beta-(2-thienyl)alanine lowered the K(i) values to (8.2 +/- 0.6) x 10(-14) or (5.1 +/- 0.4) x 10(-14) M, respectively. The linker Gly-Gly-Gly-betaAla of the inhibitors in the previous sentence was simplified with 12-aminododecanoic acid, resulting in further improvement of the K(i) values to (3.8 +/- 0.6) x 10(-14) or (1.7 +/- 0.4) x 10(-14) M, respectively. These K(i) values are equivalent to that of natural hirudin (2.2 x 10(-14) M), yet the size of the synthetic inhibitors (2 kD) is only one-third that of hirudin (7 kD). Two inhibitors, with L-norleucine or L-beta-(2-thienyl)alanine at the P1' residue and the improved linker of 12-aminododecanoic acid, were crystallized in complex with human alpha-thrombin. The crystal structures of these complexes were solved and refined to 2.1 A resolution. The Lys(60F) side chain of thrombin moved significantly and formed a large nonpolar S1' subsite to accommodate the bulky P1' residue. PMID- 10694408 TI - Hormone-sensitive lipase functions as an oligomer. AB - Hormone-sensitive lipase (HSL) is a cytosolic neutral lipase whose activity is regulated by reversible phosphorylation and which is thought to be the rate limiting enzyme for the mobilization of FFA from adipose tissue. In the current studies the subunit structure of HSL has been explored using sucrose gradient centrifugation and in vivo and in vitro protein-protein interactions. Evidence is provided to demonstrate that HSL exists as a functional dimer composed of homologous subunits. Dimeric HSL displayed approximately 40-fold greater activity against cholesteryl ester substrate when compared with monomeric HSL without any differences in affinity for the substrate. Truncations of HSL identified the importance of the N-terminal 300 amino acids, as well as other regions, in participating in the oligomerization of HSL. These studies support the notion that the N-terminal region of HSL represents a docking domain for protein-protein interactions and provide an additional mechanism for the posttranslational control of HSL activity in the cell via oligomerization. PMID- 10694409 TI - Probing the mechanistic role of glutamate residue in the zinc-binding motif of type A botulinum neurotoxin light chain. AB - Type A botulinum neurotoxin (BoNT/A) is a zinc endopeptidase that contains the consensus sequence HEXXH (residues 223-227) in the toxic light chain (LC). The X ray structure of the toxin has predicted that the two histidines of this motif are two of the three zinc-coordinating ligands and that the glutamate is a crucial amino acid involved in catalysis. The functional implication of E224 in the motif of LC was investigated by replacing the residue with glutamine and aspartate using site-directed mutagenesis. Substitution of Glu-224 with Gln (E224Q) resulted in a total loss of the endopeptidase activity, whereas substitution with Asp (E224D) retained about 1.4% of the enzymatic activity (k(cat) 140 vs 1.9 min(-1), respectively). However, K(m) values for wild-type and E224D BoNT/A LC were similar, 42 and 50 microM, respectively. Global structure, in terms of secondary structure content and topography of aromatic amino residues, Zn(2+) content, and substrate binding ability are retained in the enzymatically inactive mutants. Titration of Zn(2+) to EDTA-treated wild-type and mutant proteins indicated identical enthalpy for Zn(2+) binding. These results suggest an essential and direct role of the carboxyl group of Glu-224 in the hydrolysis of the substrate. The location of the carboxyl group at a precise position is critical for the enzymatic activity, as replacement of Glu-224 with Asp resulted in almost total loss of the activity. PMID- 10694410 TI - Effect of pressure on deuterium isotope effects of yeast alcohol dehydrogenase: evidence for mechanical models of catalysis. AB - Moderate pressure accelerates hydride transfer catalyzed by yeast alcohol dehydrogenase, indicative of a large negative volume of activation [Cho and Northrop (1999) Biochemistry 38, 7470-7475]. A comparison of the effects of pressure on the oxidation of normal versus dideuteriobenzyl alcohol generates a monophasic decrease in the intrinsic isotope effect; therefore, the volume of activation for the transition-state of deuteride transfer must be even more negative, by 10.4 mL/mol. This finding appears consistent with hydrogen tunneling previously proposed for this dehydrogenase [Cha, Y., Murray, C. J., and Klinman, J. P. (1989) Science 243, 1325-1330]. However, a global fit of the primary data shows that the entire isotope effect arises from a transition-state phenomenon, unlike normal isotope effects, which arise from different vibrational frequencies in reactant states, and tunneling isotope effects, which arise from a mixture of both states. Assuming the phenomenon is tunneling, the isotopic data are consistent with a Bell tunneling correction factor of Q(H) = 12 and an imaginary frequency of nu(H) = 1220 cm(-1), the first so calculated from experimental enzymatic data. This excessively large correction factor and the large difference in the isotopic activation volumes, plus the low isotope effects at extrapolated pressures, challenge traditional applications of physical organic chemistry and transition-state theory to enzymatic catalysis. They suggest instead that something other than transition-state stabilization or tunneling is responsible for the rate acceleration, something unique to the enzymatic transition state that does not occur in nonenzymatic reactions. Arguments for the vibrational model of coupled atomic motions and the fluctuating enzyme model of protein domain motion are put forward as possible interpretations. PMID- 10694411 TI - Naturally occurring alkylresorcinols that mediate DNA damage and inhibit its repair. AB - A study of di- and trihydroxyalkylbenzenes and bis(dihydroxyalkylbenzenes) revealed that several compounds were capable of both mediating Cu(2+)-dependent DNA cleavage and strongly inhibiting DNA polymerase beta. The most potent DNA polymerase beta inhibitors were bis(dihydroxyalkylbenzenes) 5 and 6; compounds 3 and 4 were also reasonably potent. The length of the alkyl substituent was found to be a critical element for DNA polymerase beta inhibition, since compounds 1 and 2 had shorter substituents than 3 and were completely inactive. Lineweaver Burk plots revealed that 3, 4, and 6 exhibited mixed inhibition of DNA polymerase beta with respect to both activated DNA and dTTP. Unsaturated bis(dihydroxyalkylbenzene) 5 was a pure noncompetitive inhibitor with respect to both substrates and associated avidly with the enzyme whether or not it was in complex with its substrate(s). Copper(II)-mediated DNA cleavage was the most pronounced for the trihydroxyalkylbenzene 3, consistent with an earlier report [Singh, U. S., Scannell, R. T., An, H., Carter, B. J., and Hecht, S. M. (1995) J. Am. Chem. Soc. 117, 12691-12699]. Unsaturated bis(dihydroxyalkylbenzene) 5 was the next most active DNA cleaving agent, followed by the dihydroxyalkylbenzene 4. The saturated bis(dihydroxyalkylbenzene) (6) did not cleave DNA well in a cell free system under the conditions studied but nonetheless potentiated the effects of bleomycin to the greatest extent in cell culture studies. Interestingly, compound 5 produced a reduction in the numbers of viable cells when incubated in the presence of bleomycin and a further reduction in the numbers of viable cells in the presence of both bleomycin and Cu(2+). The same effect was noted to a lesser extent for compound 3 but not for 4 or 6. PMID- 10694412 TI - Thermodynamics of substrate binding to the chaperone SecB. AB - The thermodynamics of binding of unfolded polypeptides to the chaperone SecB was investigated in vitro by isothermal titration calorimetry and fluorescence spectroscopy. The substrates were reduced and carboxamidomethylated forms of RNase A, BPTI, and alpha-lactalbumin. SecB binds both fully unfolded RNase A and BPTI as well as compact, partially folded disulfide intermediates of alpha lactalbumin, which have 40-60% of native secondary structure. The heat capacity changes observed on binding the reduced and carboxamidomethylated forms of alpha lactalbumin, BPTI, and RNase A were found to be -0.10, -0.29, and -0.41 kcal mol( 1) K(-1), respectively, and suggest that between 7 and 29 residues are buried upon substrate binding to SecB. In all cases, binding occurs with a stoichiometry of one polypeptide chain per monomer of SecB. There is no evidence for two separate types of binding sites for positively charged and hydrophobic ligands. Spectroscopic and proteolysis protection studies of the binding of SecB to poly-L Lys show that binding of highly positively charged peptide ligands to negatively charged SecB leads to charge neutralization and subsequent aggregation of SecB. The data are consistent with a model where SecB binds substrate molecules at an exposed hydrophobic cleft. SecB aggregation in the absence of substrate is prevented by electrostatic repulsion between negatively charged SecB tetramers. PMID- 10694413 TI - Refined X-ray structures of the oxidized, at 1.3 A, and reduced, at 1.17 A, PMID- 10694414 TI - Inhibition of the establishment of zygotic polarity by protein tyrosine kinase inhibitors leads to an alteration of embryo pattern in Fucus. AB - Fucoid algae, including the genus Fucus and Pelvetia, are recognized as model systems to study early embryogenesis in plants. In particular the zygotes of these fucoid algae are highly suitable experimental systems for investigating the establishment of polarity and its requirement for later embryogenesis. However, the transduction pathways involved in the initiation of polarization are still poorly understood, and the link between the early polarization processes and embryo long-term patterning has never been experimentally demonstrated. We, therefore, have investigated the putative role of protein phosphorylation in the regulation of early embryogenesis, using a combined pharmacological and biochemical approach. Among the various protein kinase inhibitors tested, a subset of well-known PTK inhibitors, including genistein, prevented germination but had no effect on growth of germinated zygotes and embryos. Inhibition of germination appeared to be a direct consequence of prevention of polarization since genistein and other PTK inhibitors specifically inhibited axis formation in a light-independent manner. Genistein inhibited cellular events associated with polarization such as polarized secretion of cell wall sulfated compounds. Anchorage of F-actin at the rhizoid pole was also inhibited and F-actin redistributed in response to a new light vector. Zygotes inhibited in the polarization process over the period of axis formation recovered from the treatment and displayed differentiated cellular structures after a few days. However, they exhibited a deeply disorganized pattern, suggesting that the early polarization process is essential for normal patterning of the embryo. Western blot analysis of protein phosphorylation showed that the patterns of protein phosphorylation changed during development and were disturbed by treatments with genistein. This drug also inhibited in vitro autophosphorylation. The nature of the genistein-sensitive kinases required for polarization and long-term patterning is discussed in light of these data. PMID- 10694415 TI - The hindsight gene is required for epithelial maintenance and differentiation of the tracheal system in Drosophila. AB - During animal development, morphogenesis of tissues and organs requires dynamic cell shape changes and movements that are accomplished without loss of epithelial integrity. Data from vertebrate and invertebrate systems have implicated several cell surface and cytoskeleton-associated molecules in the establishment and maintenance of epithelial architecture, but there has been little analysis of the genetic regulatory hierarchies that control epithelial morphogenesis in specific tissues. Here we show that the Drosophila Hindsight nuclear zinc-finger protein is required during tracheal morphogenesis for the maintenance of epithelial integrity and assembly of apical extracellular structures known as taenidia. In hindsight (hnt) mutants tracheal placodes form, invaginate, and undergo primary branching as well as early fusion events. Starting at midembryogenesis, however, the tracheal epithelium collapses or expands to give rise to sacs of tissue. While a subset of hnt mutant tracheal cells enters the apoptotic pathway, genetic suppression of apoptosis indicates that this is not the cause of the epithelial defects. Surviving hnt mutant tracheal cells retain cell-cell junctions and a normal subcellular distribution of apical markers such as Crumbs and DE-Cadherin. However, taenidia do not form on the lumenal surface of tracheal cells. While loss of epithelial integrity is a common feature of crumbs, stardust, and hnt mutants, defective assembly of taenidia is unique to hnt mutants. These data suggest that HNT is a tissue-specific factor that regulates maintenance of the tracheal epithelium as well as differentiation of taenidia. PMID- 10694416 TI - Autocrine expression and ontogenetic functions of the PACAP ligand/receptor system during sympathetic development. AB - The superior cervical ganglion (SCG) is a well-characterized model of neural development, in which several regulatory signals have been identified. Vasoactive intestinal peptide (VIP) has been found to regulate diverse ontogenetic processes in sympathetics, though functional requirements for high peptide concentrations suggest that other ligands are involved. We now describe expression and functions of pituitary adenylate cyclase-activating polypeptide (PACAP) during SCG ontogeny, suggesting that the peptide plays critical roles in neurogenesis. PACAP and PACAP receptor (PAC(1)) mRNA's were detected at embryonic days 14.5 (E14.5) through E17.5 in vivo and virtually all precursors exhibited ligand and receptor, indicating that the system is expressed as neuroblasts proliferate. Exposure of cultured precursors to PACAP peptides, containing 27 or 38 residues, increased mitogenic activity 4-fold. Significantly, PACAP was 1000-fold more potent than VIP and a highly potent and selective antagonist entirely blocked effects of micromolar VIP, consistent with both peptides acting via PAC(1) receptors. Moreover, PACAP potently enhanced precursor survival more than 2-fold, suggesting that previously defined VIP effects were mediated via PAC(1) receptors and that PACAP is the more significant developmental signal. In addition to neurogenesis, PACAP promoted neuronal differentiation, increasing neurite outgrowth 4-fold and enhancing expression of neurotrophin receptors trkC and trkA. Since PACAP potently activated cAMP and PI pathways and increased intracellular Ca(2+), the peptide may interact with other developmental signals. PACAP stimulation of precursor mitosis, survival, and trk receptor expression suggests that the signaling system plays a critical autocrine role during sympathetic neurogenesis. PMID- 10694417 TI - Analyses of segment-specific expression of alkaline phosphatase activity in the mesoderm of the oligochaete annelid Tubifex: implications for specification of segmental identity. AB - In the embryos of the oligochaete annelid Tubifex, segments VII and VIII specifically express mesodermal alkaline phophatase (ALP) activity in the ventrolateral region. In this study, we examined whether this segment-specific expression of ALP activity depends on external cues. Cell lineage analyses show that the ALP-expressing cells originate from M teloblasts. Furthermore, a set of teloblast-ablation experiments demonstrated that the seventh and eighth primary m blast cells (m7 and m8) produced from M teloblasts give rise to ALP-expressing cells in segments VII and VIII, respectively, and that primary m blast cells other than m7 and m8 lack the ability to generate ALP-expressing progeny cells. The results of another set of blastomere-ablation experiments suggest that ALP expressing cells emerge independently of interactions with surrounding tissues. Teloblast-transplantation experiments demonstrated that m8 can generate ALP expressing cells in an ectopical position, suggesting that it is unlikely that ALP activity emerges in response to the positional cues residing in the embryo. These results suggest that m7 and m8 are exclusively specified as precursors of ALP-expressing cells at the time of their birth from M teloblasts. We propose that segmental identities in primary m blast cells of the Tubifex embryo are determined according to the genealogical position in the M lineage and that the M teloblast possesses a developmental program through which the sequence of blast cell identities is determined. PMID- 10694418 TI - Influence of FGF4 on digit morphogenesis during limb development in the mouse. AB - Much of what we currently know about digit morphogenesis during limb development is deduced from embryonic studies in the chick. In this study, we used ex utero surgical procedures to study digit morphogenesis during mouse embryogenesis. Our studies reveal some similarities; however, we have found considerable differences in how the chick and the mouse autopods respond to experimentation. First, we are not able to induce ectopic digit formation from interdigital cells as a result of wounding or TGFbeta-1 application in the mouse, in contrast to what is observed in the chick. Second, FGF4, which inhibits the formation of ectopic digits in the chick, induces a digit bifurcation response in the mouse. We demonstrate with cell marking studies that this bifurcation response results from a reorganization of the prechondrogenic tip of the digit rudiment. The FGF4 effect on digit morphogenesis correlates with changes in the expression of a number of genes, including Msx1, Igf2, and the posterior members of the HoxD cluster. In addition, the bifurcation response is digit-specific, being restricted to digit IV. We propose that FGF4 is an endogenous signal essential for skeletal branching morphogenesis in the mouse. This work stresses the existence of major differences between the chick and the mouse in how digit morphogenesis is regulated and is thus consistent with the view that vertebrate digit evolution is a relatively recent event. Finally, we discuss the relationship between the digit IV bifurcation restriction and the placement of the metapterygial axis in the evolution of the tetrapod limb. PMID- 10694419 TI - BMPs are required at two steps of limb chondrogenesis: formation of prechondrogenic condensations and their differentiation into chondrocytes. AB - Formation of the long bones requires a cartilage template. Cartilage formation (chondrogenesis) proceeds through determination of cells and their aggregation into prechondrogenic condensations, differentiation into chondrocytes, and later maturation. Several studies indicate that members of the bone morphogenetic protein (BMP) family promote cartilage formation, but the exact step(s) in which BMPs are involved during this process remains undefined. To resolve this issue, we have used a retroviral vector to misexpress the BMP antagonist Noggin in the embryonic chick limb. Unlike previous reports, we have characterized the resulting phenotype in depth, analyzing histological and early chondrogenic markers, as well as the patterns of cell death and proliferation. Misexpression of Noggin prior to the onset of chondrogenesis leads to the total absence of skeletal elements, as previously reported (J. Capdevila and R. L. Johnson, 1998, Dev. Biol. 197, 205-217). Noggin inhibits cartilage formation at two distinct steps. First, we demonstrate that mesenchymal cells do not aggregate into prechondrogenic condensations, and additional results suggest that these cells persist in an undifferentiated state. Second, we show that differentiation of chondroprogenitors into chondrocytes can also be blocked, concurrent with expanded expression of a presumptive joint region marker. In addition, we observed alterations in muscle and tendon morphogenesis, and the potential role of BMPs in these processes will be discussed. Our studies therefore provide in vivo evidence that BMPs are necessary for different steps of chondrogenesis: chondroprogenitor determination and/or condensation and subsequent differentiation into chondrocytes. PMID- 10694420 TI - Reduced Pax2 gene dosage increases apoptosis and slows the progression of renal cystic disease. AB - The murine cpk mouse develops a rapid-onset polycystic kidney disease (PKD) with many similarities to human PKD. During kidney development, the transcription factor Pax2 is required for the specification and differentiation of the renal epithelium. In humans, Pax2 is also expressed in juvenile cystic kidneys where it correlates with cell proliferation. In this report, Pax2 expression is demonstrated in the cystic epithelium of the mouse cpk kidneys. To assess the role of Pax2 during the development of polycystic kidney disease, the progression of renal cysts was examined in cpk mutants carrying one or two alleles of Pax2. Reduced Pax2 gene dosage resulted in a significant inhibition of renal cyst growth while maintaining more normal renal structures. The inhibition of cyst growth was not due to reduced proliferation of the cystic epithelium, rather to increased cell death in the Pax2 heterozygotes. Increased apoptosis with reduced Pax2 gene dosage was also observed in normal developing kidneys. Thus, increased cell death is an integral part of the Pax2 heterozygous phenotype and may be the underlying cause of Pax gene haploinsufficiency. That the cystic epithelium requires Pax2 for continued expansion underscores the embryonic nature of the renal cystic cells and may provide new insights toward growth suppression strategies. PMID- 10694421 TI - Protein kinase expression during murine mammary development. AB - The susceptibility of the mammary gland to carcinogenesis is influenced by its normal development, particularly during developmental stages such as puberty and pregnancy that are characterized by marked changes in proliferation and differentiation. Protein kinases are important regulators of proliferation and differentiation, as well as of neoplastic transformation, in a wide array of tissues, including the breast. Using a RT-PCR-based cloning strategy, we have identified 41 protein kinases that are expressed in breast cancer cell lines and in the murine mammary gland during development. The expression of each of these kinases was analyzed throughout postnatal mammary gland development as well as in a panel of mammary epithelial cell lines derived from distinct transgenic models of breast cancer. Although the majority of protein kinases isolated in this screen have no currently recognized role in mammary development, most kinases examined were found to exhibit developmental regulation. After kinases were clustered on the basis of similarities in their temporal expression profiles during mammary development, multiple distinct patterns of expression were observed. Analysis of these patterns revealed an ordered set of expression profiles in which successive waves of kinase expression occur during development. Interestingly, several protein kinases whose expression has previously been reported to be restricted to tissues other than the mammary gland were isolated in this screen and found to be expressed in the mammary gland. In aggregate, these findings suggest that the array of kinases participating in the regulation of normal mammary development is considerably broader than currently appreciated. PMID- 10694422 TI - Both nuclear and cytoplasmic components are defective in oocytes of the B6.Y(TIR) sex-reversed female mouse. AB - In the mammalian gonadal primordium, activation of the Sry gene on the Y chromosome initiates a cascade of genetic events leading to testicular organization whereas its absence results in ovarian differentiation. An exception occurs when the Y chromosome of Mus musculus domesticus from Tirano, Italy (Y(TIR)), is placed on the C57BL/6J (B6) genetic background. The B6.Y(TIR) progeny develop only ovaries or ovotestes despite Sry transcription in fetal life. Consequently, the XY offspring with bilateral ovaries develop into apparently normal females, but their eggs fail to develop after fertilization. Our previous studies have shown that the primary cause of infertility can be attributed to oocytes rather than their surrounding somatic cells in the XY ovary. This study attempted to identify the defects in oocytes from the B6.Y(TIR) female mouse. We examined the developmental potential of embryos from XY and XX females after exchanging their nuclear components by microsurgery following in vitro maturation and fertilization. The results suggest that both nuclear and cytoplasmic components are defective in oocytes from XY females. In the XY fetal ovary, most germ cells entered meiosis and their autosomes appeared to synapse normally while the X and Y chromosomes remained unpaired during meiotic prophase. This lack of X-Y pairing probably caused aneuploidy in some secondary oocytes following in vitro maturation. However, normal numbers of chromosomes in the rest of the secondary oocytes indicate that aneuploidy alone can not explain the nuclear defect in oocytes. PMID- 10694423 TI - Failure of Myf5 to support myogenic differentiation without myogenin, MyoD, and MRF4. AB - The basic helix-loop-helix (bHLH) transcription factors-MyoD, Myf5, myogenin, and MRF4-can each activate the skeletal muscle-differentiation program in transfection assays. However, their functions during embryogenesis, as revealed by gene-knockout studies in mice, are distinct. MyoD and Myf5 have redundant functions in myoblast specification, whereas myogenin and either MyoD or MRF4 are required for differentiation. Paradoxically, myoblasts from myogenin mutant or MyoD/MRF4 double-mutant neonates differentiate normally in vitro, despite their inability to differentiate in vivo, suggesting that the functions of the myogenic bHLH factors are influenced by the cellular environment and that the specific myogenic defects observed in mutant mice do not necessarily reflect essential functions of these factors. Understanding the individual roles of these factors is further complicated by their ability to cross-regulate one another's expression. To investigate the functions of Myf5 in the absence of contributions from other myogenic bHLH factors, we generated triple-mutant mice lacking myogenin, MyoD, and MRF4. These mice appear to contain a normal number of myoblasts, but in contrast to myogenin or MyoD/MRF4 mutants, differentiated muscle fibers fail to form in vivo and myoblasts from neonates of this triple mutant genotype are unable to differentiate in vitro. These results suggest that physiological levels of Myf5 are insufficient to activate the myogenic program in the absence of other myogenic factors and suggest that specialized functions have evolved for the myogenic bHLH factors to switch on the complete program of muscle gene expression. PMID- 10694424 TI - p27(Kip1) regulates cell cycle withdrawal of late multipotent progenitor cells in the mammalian retina. AB - The cyclin-dependent kinase inhibitor protein, p27(Kip1), is necessary for the timing of cell cycle withdrawal that precedes terminal differentiation in oligodendrocytes of the optic nerve. Although p27(Kip1) is widely expressed in the developing central nervous system, it is not known whether this protein has a similar role in neuronal differentiation. To address this issue, we have examined the expression and function of p27(Kip1) in the developing retina, a well characterized part of the central nervous system. p27(Kip1) is expressed in a pattern coincident with the onset of differentiation of most retinal cell types. In vitro analyses show that p27(Kip1) accumulation in retinal cells correlates with cell cycle withdrawal and differentiation, and when overexpressed, p27(Kip1) inhibits proliferation of the progenitor cells. Furthermore, the histogenesis of photoreceptors and Muller glia is extended in the retina of p27(Kip1)-deficient mice. Finally, we examined the adult retinal dysplasia in p27(Kip1)-deficient mice with cell-type-specific markers. Contrary to previous suggestions that the dysplasia is caused by excess production of photoreceptors, we suggest that the dysplasia is due to the displacement of reactive Muller glia into the layer of photoreceptor outer segments. These results demonstrate that p27(Kip1) is part of the molecular mechanism that controls the decision of multipotent central nervous system progenitors to withdraw from the cell cycle. Second, postmitotic Muller glia have a novel and intrinsic requirement for p27(Kip1) in maintaining their differentiated state. PMID- 10694425 TI - P granules in the germ cells of Caenorhabditis elegans adults are associated with clusters of nuclear pores and contain RNA. AB - The germ cells, and germ cell precursors, in the nematode Caenorhabditis elegans contain distinctive granules called P granules. During early embryogenesis, P granules are segregated asymmetrically into those blastomeres that eventually produce the germ line. Because of the correlation between P granule distribution and the development of the germ line, P granules are widely thought to function in some aspect of germ line specification or differentiation. Most of the analysis of P granule structure and localization has focused on the early embryo, when P granules are located in the cytoplasm. However, during most of development P granules are associated with germ cell nuclei. We report here an ultrastructural analysis of the nuclear-associated P granules in the germ cells of the adult hermaphrodite gonad. We show that P granules are tightly associated with nuclear pores and that the positions of certain structures within the P granules correspond to the positions of pores on the nuclear envelope. We present immunocytochemical and ultrastructural data suggesting that P granules can associate, or remain associated, with pore-like structures even after they detach from the nuclear envelope during oogenesis. Finally, we show that nuclear associated P granules in the gonad contain RNA, complementing previous studies showing that cytoplasmic P granules in embryos contain RNA. PMID- 10694426 TI - The Golgi apparatus segregates from the lysosomal/acrosomal vesicle during rhesus spermiogenesis: structural alterations. AB - The acrosome is an acidic secretory vesicle containing hydrolytic enzymes that are involved in the sperm's passage across the zona pellucida. Imaging of the acrosomal vesicle and the Golgi apparatus in live rhesus monkey spermatids was accomplished by using the vital fluorescent probe LysoTracker DND-26. Concurrently, the dynamics of living spermatid mitochondria was visualized using the specific probe MitoTracker CMTRos and LysoTracker DND-26 detected the acrosomal vesicle from its formation through spermatid differentiation. LysoTracker DND-26 also labeled the Golgi apparatus in spermatogenic cells. In spermatocytes the Golgi is spherical and, in round spermatids, it is localized over the acrosomal vesicle, as confirmed by using polyclonal antibodies against Golgin-95/GM130, Golgin-97, and Golgin-160. Using both live LysoTracker DND-26 imaging and Golgi antibodies, we found that the Golgi apparatus is cast off from the acrosomal vesicle and migrates toward the sperm tail in elongated spermatids. The Golgi is discarded in the cytoplasmic droplet and is undetectable in mature ejaculated spermatozoa. The combined utilization of three vital fluorescent probes (Hoechst 33342, LysoTracker DND-26, and MitoTracker CMTRos) permits the dynamic imaging of four organelles during primate spermiogenesis: the nucleus, the mitochondria, the acrosomal vesicle, and the Golgi apparatus. PMID- 10694427 TI - Two distinct cell populations in the floor plate of the zebrafish are induced by different pathways. AB - The floor plate is a morphologically distinct structure of epithelial cells situated along the midline of the ventral spinal cord in vertebrates. It is a source of guidance molecules directing the growth of axons along and across the midline of the neural tube. In the zebrafish, the floor plate is about three cells wide and composed of cuboidal cells. Two cell populations can be distinguished by the expression patterns of several marker genes, including sonic hedgehog (shh) and the fork head-domain gene fkd4: a single row of medial floor plate (MFP) cells, expressing both shh and fkd4, is flanked by rows of lateral floor plate (LFP) cells that express fkd4 but not shh. Systematic mutant searches in zebrafish embryos have identified a number of genes, mutations in which visibly reduce the floor plate. In these mutants either the MFP or the LFP cells are absent, as revealed by the analysis of the shh and fkd4 expression patterns. MFP cells are absent, but LFP cells are present, in mutants of cyclops, one-eyed pinhead, and schmalspur, whose development of midline structures is affected. LFP cells are absent, but MFP cells are present, in mutants of four genes, sonic you, you, you-too, and chameleon, collectively called the you-type genes. This group of mutants also shows defects in patterning of the paraxial mesoderm, causing U- instead of V-shaped somites. One of the you-type genes, sonic you, was recently shown to encode the zebrafish Shh protein, suggesting that the you-type genes encode components of the Shh signaling pathway. It has been shown previously that in the zebrafish shh is required for the induction of LFP cells, but not for the development of MFP cells. This conclusion is supported by the finding that injection of shh RNA causes an increase in the number of LFP, but not MFP cells. Embryos mutant for iguana, detour, and umleitung share the lack of LFP cells with you-type mutants while somite patterning is not severely affected. In mutants that fail to develop a notochord, MFP cells may be present, but are always surrounded by LFP cells. These data indicate that shh, expressed in the notochord and/or the MFP cells, induces the formation of LFP cells. In embryos doubly mutant for cyclops (cyc) and sonic you (syu) both LFP and MFP cells are deleted. The number of primary motor neurons is strongly reduced in cyc;syu double mutants, while almost normal in single mutants, suggesting that the two different pathways have overlapping functions in the induction of primary motor neurons. PMID- 10694428 TI - Growth cones utilize both widespread and local directional cues in the zebrafish brain. AB - The distribution of cues that provide directional information for specific growth cones in the zebrafish brain was functionally assayed by transplanting epiphysial neurons to ectopic locations in the embryonic brain followed by determining the pathways taken by the donor axons. Epiphysial axons normally first extend ventrally from their position in the dorsal diencephalon and then turn and extend anteriorly in the ventral diencephalon. When transplanted to ectopic sites at other axial levels of the brain, where in principle the axons could extend in any direction, epiphysial axons consistently extended ventrally. Furthermore, following initial ventral extension ectopic epiphysial axons turned randomly in the anterior and posterior directions. These results suggest that the cues for ventral extension are widely distributed along the rostrocaudal axis of the zebrafish brain, but the cues for subsequent anterior extension are restricted to the site where the epiphysial axons normally turn longitudinally. PMID- 10694429 TI - Fate mapping of the mouse prosencephalic neural plate. AB - Little is known about the behavior of cells within the anterior neural plate or tube in developing mammalian embryos in utero due to technical limitations. Here we labeled neuroepithelial cells with vital dye and traced their siblings for 1 or 2 days using the whole-embryo culture system. The results demonstrated that rostral cell movement from the midbrain to the forebrain in the mouse neural plate was restricted at the boundary by the five-somite stage. Coincident with restriction of cell intermingling, expression of a transcription factor, Pax6, and a cell adhesion molecule, cadherin-6, commmenced to demarcate the forebrain compartment. Within this compartment, we also mapped several prospective regions of the telencephalon and diencephalon to the eyes. The fate map of the mouse prosencephalic neural plate was very similar to those of other vertebrates, providing evidence that mammalian-specific brain structures, represented in the cerebral neocortex, could evenly develop along the conserved framework of neuromeres. PMID- 10694430 TI - Regulation of microphthalmia-associated transcription factor MITF protein levels by association with the ubiquitin-conjugating enzyme hUBC9. AB - The basic helix-loop-helix/leucine zipper (bHLH/ZIP) microphthalmia-associated transcription factor (MITF) regulates transcription of genes encoding enzymes essential for melanin biosynthesis in melanocytes and retinal pigmented epithelial cells. To determine how MITF activity is regulated, we used the yeast two-hybrid system to identify proteins expressed by human melanoma cells that interact with MITF. The majority of clones that showed positive interaction with a 158-amino-acid region of MITF containing the bHLH/ZIP domain (aa 168-325) encoded the ubiquitin conjugating enzyme hUBC9. The association of MITF with hUBC9 was further confirmed by an in vitro GST pull-down assay. Although hUBC9 is known to interact preferentially with SENTRIN/SUMO1, in vitro transcription/translation analysis demonstrated greater association of MITF with ubiquitin than with SENTRIN. Importantly, cotransfection of MITF and hUBC9 expression vectors resulted in MITF protein degradation. MITF protein was stabilized by the proteasome inhibitor MG132, indicating the role of the ubiquitin-proteasome system in MITF degradation. Serine 73, which is located in a region rich in proline, glutamic acid, serine, and threonine (PEST), regulates MITF protein stability, since a serine to alanine mutation prevented hUBC9 mediated MITF (S73A) degradation. Furthermore, we identified lysine 201 as a potential ubiquitination site. A lysine to arginine mutation abolished MITF (K201R) degradation by hUBC9 in vivo. Our experiments indicate that by targeting MITF for proteasome degradation, hUBC9 is a critical regulator of melanocyte differentiation. PMID- 10694431 TI - Activation of a cysteine protease in MCF-7 and T47D breast cancer cells during beta-lapachone-mediated apoptosis. AB - beta-Lapachone (beta-lap) effectively killed MCF-7 and T47D cell lines via apoptosis in a cell-cycle-independent manner. However, the mechanism by which this compound activated downstream proteolytic execution processes were studied. At low concentrations, beta-lap activated the caspase-mediated pathway, similar to the topoisomerase I poison, topotecan; apoptotic reactions caused by both agents at these doses were inhibited by zVAD-fmk. However at higher doses of beta lap, a novel non-caspase-mediated "atypical" cleavage of PARP (i.e., an approximately 60-kDa cleavage fragment) was observed. Atypical PARP cleavage directly correlated with apoptosis in MCF-7 cells and was inhibited by the global cysteine protease inhibitors iodoacetamide and N-ethylmaleimide. This cleavage was insensitive to inhibitors of caspases, granzyme B, cathepsins B and L, trypsin, and chymotrypsin-like proteases. The protease responsible appears to be calcium-dependent and the concomitant cleavage of PARP and p53 was consistent with a beta-lap-mediated activation of calpain. beta-Lap exposure also stimulated the cleavage of lamin B, a putative caspase 6 substrate. Reexpression of procaspase-3 into caspase-3-null MCF-7 cells did not affect this atypical PARP proteolytic pathway. These findings demonstrate that beta-lap kills cells through the cell-cycle-independent activation of a noncaspase proteolytic pathway. PMID- 10694432 TI - Effects of keratin filament disruption on exocrine pancreas-stimulated secretion and susceptibility to injury. AB - Disruption or absence of hepatocyte keratins 8 and 18 is associated with chronic hepatitis, marked hepatocyte fragility, and a significant predisposition to stress-induced liver injury. In contrast, pancreatic keratin disruption in transgenic mice that express keratin 18 Arg89 --> Cys (K18C) is not associated with an obvious pancreatic pathology. We compared the effects of keratin filament disruption on pancreatic acini or acinar cell viability, and on cholecystokinin (CCK)-stimulated secretion, in transgenic mice that overexpress wild-type keratin 18 and harbor normal extended keratin filaments (TG2) and K18C mice. We also compared the response of these mice to pancreatitis induced by a choline deficient ethionine-supplemented diet or by caerulein. Despite extensive cytoplasmic keratin filament disruption, the apicolateral keratin filament bundles appear intact in the acinar pancreas of K18C mice, as determined ultrastructurally and by light microscopy. No significant pancreatitis-associated histologic, serologic, or F-actin/keratin apicolateral redistribution differences were noted between TG2 and K18C mice. Acinar cell viability and yield after collagenase digestion were lower in K18C than in TG2 mice, but the yields of intact acini and their (125)I-CCK uptake and responses to CCK-stimulated secretion were similar. Our results indicate that keratin filament reorganization is a normal physiologic response to pancreatic cell injury, but an intact keratin cytoplasmic filament network is not as essential in protection from cell injury as in the liver. These findings raise the possibility that the abundant apicolateral acinar keratin filaments, which are not as evident in hepatocytes, may play the cytoprotective role that is seen in liver and other tissues. Alternatively, identical keratins may function differently in different tissues. PMID- 10694433 TI - Increased activity of the protein kinase C-delta holoenzyme in the cytoplasmic particulate fraction precedes the activation of caspases in polyomavirus transformed pyF111 rat fibroblasts exposed to calphostin C or topoisomerase-II inhibitors. AB - A caspase-mediated release of the 40-kDa catalytic fragment of the delta isoform (CF-delta) of protein kinase C (PKC-delta) is involved in apoptosis, but its actual role in apoptosis development is still unknown. In an effort to understand this role, we have used polyomavirus-transformed pyF111 rat fibroblasts, which are hypersusceptible to apoptosis as they constitutively hyperexpress PKC-delta, but cannot make the antiapoptotic Bcl-2 and Bcl-X(L) proteins, while making the proapoptotic Bax protein. Calphostin C is reportedly both a specific inhibitor of PKC-delta activity (C. Keenan, N. Goode, and C. Pears, 1997, FEBS Lett. 415, 101 108) and an effective apoptogen (M. Murata et al., 1997, Cell. Mol. Life Sci. 53, 737-743). Exposure of pyF111 cells to calphostin C (75 nM) stimulated the translocation of the PKC-delta holoenzyme (holo-PKC-delta) onto the cytoplasmic particulate (CP) fraction between 15 and 45 min, which was after the release of mitochondrial cytochrome c but before the activation of cytoplasmic DEVD-specific caspases. The CF-delta fragment started accumulating only between 2 and 4 h, while apoptosis occurred mostly within 6 h. Incubating pyF111 cells with the much slower acting, apoptogenic topoisomerase-II inhibitors etoposide (VP-16) and teniposide (VM-26) also caused within 6 h a doubling of the CP-bound holo-PKC delta-related activity but with no significant translocation of the holoenzyme to the CP fraction. Again this occurred after the release of cytochrome c but before the activation of DEVDases and the accumulation of the CF-delta. However, while calphostin C did not affect the delta-related activity in the nuclear membrane (NM) and nucleoplasmic (NP) fractions, VP-16 and VM-26 caused a prompt, large, and irreversible drop in the delta activity at the NM and a transient surge followed by a fall in the NP-associated activity. Hence, a surge of CP-anchored holo-PKC-delta activity is a common part of the signals given by various apoptogenic drugs to pyF111 cells. On the other hand, inhibition of delta-related activity, first at the NM and then in the NP fraction, is a specific feature only of the signals given by apoptogenic DNA-damaging agents. PMID- 10694434 TI - Cdk1 is essential for mammalian cyclosome/APC regulation. AB - The cyclosome/APC (anaphase-promoting complex), the major component of cell-cycle specific ubiquitin-mediated proteolysis of mitotic cyclins and of other cell cycle proteins, is essential for sister chromatid separation and for exit from mitosis. Cyclosome activity and substrate specificity are modulated by phosphorylation and by transient interactions with Fizzy/cdc20 (Fzy) and Fizzy related/Hct1/Cdh1 (Fzr). This regulation has been studied so far in Drosophila embryos, in yeast, and in cell-free extracts in vitro. Studying cyclosome regulation in mammalian cells in vivo we found that both Fzr overexpression and Cdk1 inhibition can override the prometaphase checkpoint. We further show that Fzr activation of the cyclosome is negatively regulated by Cdk1. Finally, we show that the mammalian cdc14 phosphatase, like its budding yeast homologue, plays a role in cyclosome pathway regulation. These results suggest that Cdk1 is essential for coupling various activities of the cyclosome and in particular for preventing Fzr from short-circuiting the spindle pole checkpoint. Cdk1-cyclin B is thus an inhibitor, activator, and substrate of the cyclosome. PMID- 10694435 TI - Presenilin 2 expression in neuronal cells: induction during differentiation of embryonic carcinoma cells. AB - Mutations in the presenilin 1 and 2 (PS1 and PS2) genes cause most cases of early onset Alzheimer's disease. The genes encode two homologous multipass membrane proteins. Since the endogenous expression of PS2 has been poorly analyzed to date, we studied PS2 expression and localization in cultured human neuroblastoma cells and mouse neuronal cells. PS2 was mainly detected as a full-length protein of about 52 kDa in these cells and in brain, in contrast to PS1 that is mainly detected as endoproteolytic N-terminal and C-terminal fragments. Using immunofluorescence we found that like PS1, PS2 colocalized with markers of the endoplasmic reticulum-Golgi intermediate compartment, ERGIC-53 and beta-COP. Double labeling for PS1 and PS2 indicated that both proteins are colocalized in neuroblastoma SH-SY5Y cells. To study PS2 expression during differentiation, mouse embryonic carcinoma P19 cells were treated with retinoic acid. We found minimal PS2 expression in undifferentiated cells, an increase from day 2, and a maximum at day 8 after treatment. PS1 expression remained constant during this period. The differential expression of PS1 and PS2 within the P19 cells following retinoic acid treatment indicates different utilization or temporal requirements for these proteins during neuronal differentiation. PMID- 10694436 TI - Analysis of beta-catenin aggregation and localization using GFP fusion proteins: nuclear import of alpha-catenin by the beta-catenin/Tcf complex. AB - beta-Catenin plays essential roles in cell adhesion, by associating with cadherins, and as a signaling molecule, by interacting with the Tcf/LEF-1 family of transcription factors. In order to study the protein-protein interactions of beta-catenin in living cells, we fused it to green fluorescent protein (GFP). GFP beta-catenin was incorporated into cell junctions but also accumulated in the nucleus, where it formed rod-like structures. The carboxyl-terminal armadillo repeats of GFP-beta-catenin were sufficient for nuclear localization, but formation of rods required the armadillo repeats and sequences in both the amino- and the carboxyl-terminal domains. Rod formation was prevented by coexpression of N-cadherin, APC, and Tcf-4, which bind to the armadillo repeats of beta-catenin, but not by coexpression of alpha-catenin, although alpha-catenin expression did prevent accumulation of beta-catenin in the nucleus. Interestingly, when alpha catenin, beta-catenin, and Tcf-4 were coexpressed they colocalized in the nucleus, and this correlated with a decrease in beta-catenin/Tcf-dependent transcriptional activity. These results indicate that binding of beta-catenin to Tcf-4 overrides the function of alpha-catenin to sequester beta-catenin in the cytoplasm and suggest that alpha-catenin can regulate beta-catenin signaling in the nucleus. PMID- 10694437 TI - Caveolin internalization by heat shock or hyperosmotic shock. AB - We investigated the cellular localization of caveolin, a landmark protein of caveolae, by indirect immunofluorescence after heat shock or hyperosmotic shock. Caveolin was internalized to the perinucleus by heat shock (43 degrees C) and relocalized in the plasma membrane after recovery of NIH3T3 cells at 37 degrees C for 4 h. The caveolin internalization was also observed after cells were exposed to hyperosmotic shock. Caveolin disappeared from detergent-insoluble complexes in the heat-shocked cells, but alkaline phosphatase was still there, suggesting that their responses to heat shock are quite different even though both of them were enriched in detergent-insoluble complexes of normal cells. Caveolin was internalized by the actin depolymerizer cytochalasin D, but not by the tubulin depolymerizer nocodazole. In addition, cellular exposure to hydrogen peroxide caused caveolin internalization along with disintegrated microfilaments and intact microtubules. Since cellular exposure to heat shock showed disintegrated microfilaments but intact microtubules, caveolin internalization might be due to depolymerized microfilaments. When cells were exposed to heat shock and allowed to recover for 4 h, actin depolymerization and caveolin internalization were not induced by a second heat shock, suggesting that some heat shock protein(s) might prevent actin depolymerization and caveolin internalization. PMID- 10694438 TI - Spatial distribution of GC- and AT-rich DNA sequences within human chromosome territories. AB - Previous topological analyses of DNA sequence organization in the interphase chromosome mainly focused on the spatial distribution of individual gene copies within chromosome territories. In order to achieve a more comprehensive view into the subchromosomal arrangement of DNA, we isolated the GC-richest/gene-richest fraction (H3 isochores) as well as AT-richest/gene-poorest fraction of human genomic DNA (L1+L2 isochores) and visualized the respective DNA within individual chromosome territories by means of dual-color FISH. Application of confocal laser scanning microscopy and dedicated 3D image analysis software, which differentiated territory subvolumes by peeling shells one voxel in width, revealed a significant difference in the intraterritorial distribution of these two DNA sequence classes. While the H3 isochores were found localized in all subvolumes of the territories at similar frequency, simultaneously detected L1+L2 isochores were observed more to the interior of the same chromosome territories. Thus the GC-rich sequences display a much higher variability in their intraterritorial localization than AT-rich DNA fragments. PMID- 10694439 TI - Single amino acid (arginine) deprivation induces G1 arrest associated with inhibition of cdk4 expression in cultured human diploid fibroblasts. AB - Withdrawal of a single amino acid (arginine) from freely cycling early passage primary human fibroblasts caused a halt to proliferation, characterized by an accumulation of cells in the G1 phase of the cell cycle. This arrest was accompanied by the suppression of cyclin D1- and cyclin E-associated kinase activities and the appearance of hypophosphorylated retinoblastoma protein. Arginine-deprived cells remained viable for in excess of 4 days and could be made to synchronously reenter the cell cycle by restoration of the amino acid, with kinetics characteristic of exit from a quiescent state. Stimulation of cells arrested by serum withdrawal did not result in S-phase entry when arginine was omitted from the culture medium. Although cyclin D1 accumulated on normal schedule, cdk4, which increased following restimulation in amino acid-replete medium, was not induced when arginine was absent. These results suggest that arginine deprivation-in common with other "suboptimal" conditions-inhibits the passage of normal human cells through the restriction point and implicate cdk4 as the key regulatory element in amino acid-sensitive cell cycle control. PMID- 10694440 TI - Both increased stability and transcription contribute to the induction of the urokinase plasminogen activator receptor (uPAR) message by hypoxia. AB - Both hypoxia and overexpression of the urokinase plasminogen activator receptor (uPAR) are associated with a poor clinical outcome in human cancers. Hypoxia has been shown to induce uPAR expression in breast cancer cells and to increase their invasion through Matrigel, a phenomenon which can be blocked using an anti-uPAR antibody. We examined expression of uPAR mRNA in MCF7 human breast carcinoma cells under hypoxia and found that an increase in the level of the message could be detected at 1% oxygen but was most marked at 0.2 or 0.05% oxygen with an induction of 9- to 20-fold over baseline. To determine whether changes in RNA stability contributed to this dramatic increase, we used actinomycin D to inhibit transcription and found that the half-life of the message was much longer under hypoxic conditions (approximately 10 h) than during reoxygenation (approximately 2 h). Transient transfections using a luciferase reporter construct containing 2 kbp of the mouse uPAR promoter showed that promoter activity increased up to 5 fold after exposure to 0.2% oxygen. Thus, hypoxic induction of the uPAR message in MCF7 cells is due to both mRNA stabilization and increased transcriptional activation. PMID- 10694441 TI - A region containing a proline-rich motif targets sG(i2) to the golgi apparatus. AB - The central function of heterotrimeric GTP-binding proteins (G proteins) is the transduction of extracellular signals, via membrane receptors, leading to the activation of intracellular effectors. In addition to being associated with the plasma membrane, the alpha subunits of some of these proteins have also been localized in intracellular compartments. The mRNA of the G-protein inhibitory alpha subunit 2 (G(alphai2)) encodes two proteins, G(alphai2) and sG(i2), by an alternative splicing mechanism. sG(i2) differs from G(alphai2) in the C-terminal region and localizes in the Golgi in contrast to the plasma membrane localization of G(alphai2). In this paper we show that the sequence specific to sG(i2) can direct the Golgi localization of other G(alphai) subunits, but not of the stimulatory subunit G(alphas) or of a secreted protein. This indicates that, in addition to the sG(i2) C-terminus, sequences located elsewhere in the protein are required to determine the Golgi localization. Inside the sG(i2) C-terminal region we have identified a 14-amino-acid proline-rich motif which specifies the Golgi localization. Finally, we show that the sG(i2) subunit, once activated, leaves the Golgi to be localized in the endoplasmic reticulum. PMID- 10694442 TI - A C-terminal carbohydrate-binding domain in the endothelial cell regulatory protein, pigpen: new function for an EWS family member. AB - The potential for encoding information in carbohydrate (CHO) structures has long been recognized. Selective CHO-binding proteins known as lectins and the biological events they mediate are well known. However, many lectins were originally discovered for biological activities other than saccharide binding, and only subsequently was it realized that one or more of their key functions were mediated by specific CHO recognition. Our previous observations suggested that the nuclear protein pigpen had an affinity for CHO structures. This would represent a new attribute for proteins of the EWS (Ewing's sarcoma) family, of which pigpen is a member. In this study we demonstrate that a CHO-binding domain resides in the C-terminus of the molecule and can be preferentially inhibited by saccharides, most notably N-acetyl-d-galactosamine (GalNAc) and the GalNAc containing polysaccharide, chondroitin sulfate. Ligand blotting experiments were subsequently performed with fractionated, [(3)H]galactose-labeled cells to demonstrate the presence of chondroitin sulfate-inhibitable endogenous CHO ligands for pigpen in endothelial nuclei. Finally, microinjection of polysaccharide competitor into the nucleus of cultured endothelial cells resulted in a loss of pigpen focal accumulations, suggesting that the CHO-binding activity may be instrumental in subcellular localization of the protein. In summary, our results show ligand preference and domain specificity for pigpen's CHO affinity and provide initial evidence for physiological ligands and function. They may also shed new light on the mechanisms of oncogenic transformation involving EWS proteins. PMID- 10694443 TI - Galectin-3 expression and subcellular localization in senescent human fibroblasts. AB - Galectin-3 is a galactose/lactose-binding protein (M(r) approximately 30,000), identified as a required factor in the splicing of pre-mRNA. In the LG1 strain of human diploid fibroblasts, galectin-3 could be found in both the nucleus and the cytoplasm of young, proliferating cells. In contrast, the protein was predominantly cytoplasmic in senescent LG1 cells that have lost replicative competence through in vitro culture. Incubation of young cells with leptomycin B, a drug that disrupts the interaction between the leucine-rich nuclear export signal and its receptor, resulted in the accumulation of galectin-3 inside the nucleus. In senescent cells, galectin-3 staining remained cytoplasmic even in the presence of the drug, thus suggesting that the observed localization in the cytoplasm was due to a lack of nuclear import. In heterodikaryons derived from fusion of young and senescent LG1 cells, the predominant phenotype was galectin-3 in both nuclei. These results suggest that senescent LG1 cells might lack a factor(s) specifically required for galectin-3 nuclear import. PMID- 10694444 TI - Expression of the sodium/calcium exchanger in mammalian skeletal muscle cells in primary culture. AB - Previous investigations have demonstrated molecular and functional expression, at early phases of development of skeletal muscle cells in primary culture, of cardiac isoforms of proteins involved in calcium transport and regulation, like the L-type calcium channel. Here the expression of the cardiac isoform of the Na(+)/Ca(2+) exchanger (NCX1) was studied in skeletal muscle cells developing in vitro, by using biochemical, immunological, and electrophysiological techniques. Northern and Western blot experiments revealed the presence of this cardiac exchanger and its increasing expression during the early phases of development. Confocal imaging of myotubes showed an NCX1 distribution that was predominantly sarcolemmal. The whole-cell patch-clamp technique allowed us to record ionic currents, the direction and the amplitude of which depended on extracellular sodium and calcium concentrations. The developmental changes of this functional expression could be correlated with the molecular NCX1 expression changes. Taken together these data demonstrate the presence of the NCX1 isoform of the Na(+)/Ca(2+) exchanger during in vitro myogenesis and reinforce the theory that significant levels of cardiac-type proteins are transiently expressed during the early phases of the skeletal muscle cell development. PMID- 10694445 TI - The role of extracellular and cytoplasmic splice domains of alpha7-integrin in cell adhesion and migration on laminins. AB - The major laminin-binding integrin of skeletal, smooth, and heart muscle is alpha7beta1-integrin, which is structurally related to alpha6beta1. It occurs in three cytoplasmic splice variants (alpha7A, -B, and -C) and two extracellular forms (X1 and X2) which are developmentally regulated and differentially expressed in skeletal muscle. Previously, we have shown that ectopic expression of the alpha7beta-integrin splice variant in nonmotile HEK293 cells specifically induced cell locomotion on laminin-1 but not on fibronectin. To investigate the specificity and the mechanism of the alpha7-mediated cell motility, we expressed the three alpha7-chain cytoplasmic splice variants, as well as alpha6A- and alpha6B-integrin subunits in HEK293 cells. Here we show that all three alpha7 splice variants (containing the X2 domain), as well as alpha6A and alpha6B, promote cell attachment and stimulate cell motility on laminin-1 and its E8 fragment. Deletion of the cytoplasmic domain (excluding the GFFKR consensus sequence) from alpha7B resulted in a loss of the motility-enhancing effect. On laminin-2/4 (merosin), the predominant isoform in mature skeletal muscle, only alpha7-expressing cells showed enhanced motility, whereas cells transfected with alpha6A and alpha6B neither attached nor migrated on laminin-2. Adhesion of alpha7-expressing cells to both laminin-1 and laminin-2 was specifically inhibited by a new monoclonal antibody (6A11) specific for alpha7. Expression of the two extracellular splice variants alpha7X1 and alpha7X2 in HEK293 cells conferred different motilities on laminin isoforms: Whereas alpha7X2B promoted cell migration on both laminin-1 and laminin-2, alpha7X1B supported motility only on laminin-2 and not on laminin-1, although both X1 and X2 splice variants revealed similar adhesion rates to laminin-1 and -2. Fluorescence-activated cell sorter analysis revealed a dramatic reduction of surface expression of alpha6 integrin subunits after alpha7A or -B transfection; also, surface expression of alpha1-, alpha3-, and alpha5-integrins was significantly reduced. These results demonstrate selective responses of alpha6- and alpha7-integrins and of the alpha7 splice variants to laminin-1 and -2 and indicate differential roles in laminin controlled cell adhesion and migration. PMID- 10694446 TI - Differential localization of ICAD-L and ICAD-S in cells due to removal of a C terminal NLS from ICAD-L by alternative splicing. AB - CAD/CPAN/DFF40 is an apoptotic nuclease that is associated with the regulatory subunit ICAD/DFF in healthy cells. ICAD has two forms, ICAD-L/DFF45 and ICAD S/DFF35, which are transcribed from a single gene by alternative splicing. They differ at the C-terminus: 70 amino acids of ICAD-L are replaced by 4 different amino acids in ICAD-S. We previously showed that both transfected and endogenous ICAD-L are nuclear; however, the localization of ICAD and CAD remains controversial and an important issue to clarify. Here we present the evidence that ICAD-L is nuclear due to the presence of an autonomous nuclear localization signal located in the C-terminal 20 amino acids. This NLS is missing from ICAD-S, which is distributed throughout the cell. We also showed that a GFP:CAD fusion protein is located in the nucleus of transfected cells. PMID- 10694447 TI - Radiation-induced centrosome overduplication and multiple mitotic spindles in human tumor cells. AB - The centrosome is a highly regulated organelle and its proper duplication is indispensable for the formation of bipolar mitotic spindles and balanced chromosome segregation. To elucidate a possible linkage between centrosome duplication and radiation-induced nuclear damage, we examined centrosome dynamics in U2-OS osteosarcoma cells following gamma-irradiation. Nearly all control cells contained one or two centrosomes, and at mitosis more than 97% of the cells displayed typical bipolar spindles. In contrast, over 20% of cells at 48 h after 10 Gy gamma-irradiation contained more than two centrosomes, and 60% of the mitotic cells showed aberrant spindles organized by multiple poles. Remarkably, the cells with multiple centrosomes frequently exhibited changes in size and/or morphology of the nucleus, including micronuclei formation. We conclude that abnormal centrosome duplication could be one of the key events involved in nuclear fragmentation and perhaps even cell death following irradiation. PMID- 10694448 TI - Sox9 expression during chondrogenesis in micromass cultures of embryonic limb mesenchyme. AB - Sox9 plays a crucial role in chondrogenesis. It encodes an HMG-domain transcription factor that activates an enhancer in the gene for type II collagen (Col2a1), a principal cartilage matrix protein. We have characterized the temporal pattern of Sox9 RNA expression in micromass culture, a widely used in vitro model for the analysis of embryonic cartilage differentiation. Cultures were prepared from distal subridge mesenchyme of the stage 24/25 chick embryo wing bud, which undergoes uniform chondrogenic differentiation in vitro. The early "prechondrogenic" phase of culture was characterized by the activation of Sox9 RNA expression, which preceded detectable upregulation of Col2a1 transcription. Sox9 RNA levels peaked between 20 and 65 h of culture, a phase of progressive Col2a1 transcript accumulation, then declined in the mature cartilage of 120-h cultures. Staurosporine treatment enhanced chondrogenesis in micromass culture by inducing a rapid quantitative increase in Sox9 transcript levels. However, PMA, a phorbol ester that inhibits Col2a1 expression and chondrocyte differentiation, had an unexpectedly modest effect on Sox9 RNA accumulation. PMID- 10694449 TI - Editorial PMID- 10694450 TI - Antibody modeling: implications for engineering and design. AB - Our understanding of the rules relating sequence to structure in antibodies has led to the development of accurate knowledge-based procedures for antibody modeling. Information gained from the analysis of antibody structures has been successfully exploited to engineer antibody-like molecules endowed with prescribed properties, such as increased stability or different specificity, many of which have a broad spectrum of applications both in therapy and in research. Here we describe a knowledge-based procedure for the prediction of the antibody variable domains, based on the canonical structures method for the antigen binding site, and discuss its expected accuracy and limitations. The rational design of antibody-based molecules is illustrated using as an example one of the most widely employed modifications of antibody structures: the humanization of animal-derived antibodies to reduce their immunogenicity for serotherapy in humans. PMID- 10694451 TI - Automated docking of ligands to antibodies: methods and applications. AB - Many approaches to studying protein-ligand interactions by computational docking are currently available. Given the structures of a protein and a ligand, the ultimate goal of all docking methods is to predict the structure of the resulting complex. This requires a suitable representation of molecular structures and properties, search algorithms to efficiently scan the configuration space for favorable interaction geometries, and accurate scoring functions to evaluate and rank the generated orientations. For many of the available methods, tests on experimentally known antibody-antigen or antibody-hapten complexes have appeared in the literature. In addition, some of them have been used in predictive studies on antibody-ligand interactions to provide structural insights where adequate experimental information is missing. The AutoDock program is presented as example of a method for flexibly docking ligands to antibodies. Applying parameters of the second-generation AMBER force field, three antibody-hapten complexes (AN02, DB3, NC6.8) are used as new test cases to analyze the ability of the method to reproduce experimental findings. The X-ray structures could be reconstituted and the corresponding solutions were ranked with best energy score in all cases. Docking to the free instead of the complexed NC6.8 structure indicated the limits of the rigid protein treatment, although fairly good guesses about the location of the binding site and the contact residues could still be obtained if conformational flexibility was allowed at least in the ligand. PMID- 10694452 TI - Continuum electrostatic methods applied to pH-dependent properties of antibody antigen association. AB - Protein association events are a critical component of the functioning of biological systems. Antibody/antigen association, which involves extraordinarily specific interactions, has been a paradigm for the study of structural factors and intermolecular forces controlling protein-protein association. As new experimental approaches to the study of antibody/antigen affinity have become routine, and as more structures of complexes of antibodies and their antigens have become available, it has become possible to use computational approaches to study these interactions. Electrostatic interactions are known to play an important role in protein complex formation. In this review, we focus on the use of continuum electrostatic methods to compute pH-dependent properties of proteins and discuss the use of these methods in the study of antibody/antigen complexes. PMID- 10694453 TI - Experimental design for analysis of complex kinetics using surface plasmon resonance. AB - Using BIAcore surface plasmon resonance technology, we found that the real-time association kinetics of Fabs specific for hen egg-white lysozyme did not conform to a 1:1 Langmuir association model. Heterogeneity of the components is not the source of the complex kinetics. Informed by independent structural data suggesting conformational flexibility differences among these antibodies, we chose global mathematical analysis based on a two-phase model, consistent with the encounter-docking view of protein-protein associations. Experimental association times (T(a)) from 2 to 250 min revealed that initial dissociation rates decreased with increasing T(a), confirming a multiphasic association. The relationship between observed dissociation rate and T(a) is characteristic of each antibody-antigen complex. We define a new parameter, T(50), the time at which the encounter and final complexes are of equimolar concentration. The observed T(50) is a function of analyte concentration and the encounter and docking rate constants. Simulations showed that when the ligand is saturated at high analyte concentrations, T(50) reaches a minimum value, T(50)(MIN), which can be used to compare antigen-antibody complexes. For high-affinity complexes with rapid rearrangement to a stable complex, T(50)(MIN) approaches T(1/2) of the rearrangement forward rate constant. We conclude that experiments with a range of T(a) are essential to assess the nature of the kinetics, regardless of whether a two-state or 1:1 model is applicable. We suggest this strategy because each T(a) potentially reveals a different distribution of molecular states; for two-step analysis, a range of T(a) that brackets T(50) is optimal. PMID- 10694454 TI - Application of surface plasmon resonance toward studies of low-molecular-weight antigen-antibody binding interactions. AB - Methods for studying low-molecular-weight antigen-antibody binding interactions using surface plasmon resonance detection are presented. The experimental parameters most relevant to studies of low-molecular-weight antigen-antibody binding interactions are discussed. Direct kinetic analysis of the binding interactions is most informative, providing both apparent association and dissociation rate constants from which equilibrium constants can be calculated. Equilibrium analysis, including steady-state and solution affinity studies, offers an alternative approach to direct kinetic analysis when knowledge of the individual kinetic rate constants is not required or difficult to determine. The various methods are illustrated by studies of an anti-T(4) Fab fragment binding interaction with several thyroxine analogs. The methods utilized were dependent on the affinity of the interaction. The high-affinity anti-T(4) Fab fragment/l T(4) binding interaction was evaluated using direct kinetic analysis. An intermediate affinity anti-T(4) Fab fragment/l-T(3) binding interaction was evaluated using a combination of direct kinetic analysis, steady-state analysis, and solution affinity analysis. The relatively weak anti-T(4) Fab fragment/l-T(2) binding interaction was evaluated using steady-state and solution affinity analysis protocols. Several thyroxine tracers that could not be immobilized to a biosensor surface were also evaluated via the solution affinity format. In cases where a given binding interaction was examined using multiple methods the results were comparable. PMID- 10694455 TI - Force probe measurements of antibody-antigen interactions. AB - The surface force apparatus has been used to quantify directly the forces that govern the interactions between proteins and ligands. In this work, we describe the measured interactions between the antigen fluorescein and the Fab' fragment of the monoclonal 4-4-20 anti-fluorescyl IgG antibody. Here we first describe the use of the surface force apparatus to demonstrate directly the impact of the charge composition in the region of the antibody binding site on the antibody interactions. Several approaches are described for immobilizing antigens, antibodies, and proteins in general for direct force measurements. The measured force profiles presented are accompanied by an extensive discussion of protocols used to analyze the force-distance curves and to interpret them in terms of the antibody structure. In addition to long-range electrostatic forces, we also consider short-range forces that can affect the strength of adhesion between the Fab' and immobilized fluorescein. The latter investigations demonstrate the influence of interfacial properties on the recognition of surface-bound antigens. PMID- 10694456 TI - Optical spectroscopy in studies of antibody-hapten interactions. AB - This article describes the use of optical spectroscopy in studying antibody hapten interactions and in determining the equilibrium binding constants. Along with equilibrium binding data, spectroscopic tools often deliver structural information on binding-induced conformational changes of antibodies (or haptens). Structural implications of results from example antibody-hapten systems are included. Fluorescence spectroscopy has been particularly useful in the area of ligand binding, and thus steady-state fluorescence quenching and fluorescence polarization are the primary techniques under discussion. A brief description of fluorescence correlation spectroscopy is also provided. Absorption techniques, including circular dichroism, are mentioned to a lesser extent. A basic description of the mathematical models involved in the analysis of binding equilibria is provided along with references to more complete works. Simulated and experimental data are used to illustrate the various experimental protocols and the appropriate analytical methods. Typical sources of errors and experimental precautions are indicated throughout the general discussion. PMID- 10694457 TI - Analysis of correlated motion in antibody combining sites from molecular dynamics simulations. AB - The crystal structures of the NC6.8-antisweet taste ligand complex and the uncomplexed antibody structures display significant differences in the conformations of residues in the combining site. A molecular dynamics method was employed to understand the flexibility and correlated motion of key combining site residues in the uncomplexed antibody. The simulations reveal that residues that show conformational differences between the complex and uncomplexed structures display strong dynamical correlations. Extensive analysis of the dynamics trajectory using time correlation methods is presented. PMID- 10694458 TI - Synthetic haptens as probes of antibody response and immunorecognition. AB - The molecular forces that bind antibody to antigen have long fascinated chemists. The use of synthetic haptens to study immunochemical phenomena can be traced back to the classic work of Karl Lansteiner. His utilization of small-molecule-protein conjugates first demonstrated the shape-selective nature of antibody binding. Later work by Linus Pauling and David Pressman employed multivalent, synthetic ligands to establish the bivalent nature of antibodies and explain the nature of immunoprecipitation. Fluorescent probes such as dansyl, fluorescein, and Ru(bpy)(2+)(3) have been used to study affinity maturation, quantify antibody affinities, and investigate polyclonal antibody heterogeneity. Finally, X-ray crystallography has yielded a molecular picture of how antibodies exercise intermolecular forces (e.g., charge-charge interactions, H-bonding, and Van der Waals) to bind haptens. Studies inspired by Landsteiner's original work continue to play an important role in fields ranging from immunodiagnostics to catalytic antibodies. PMID- 10694459 TI - Neuromagnetic localization of CMV generators using incomplete and full-head biomagnetometer. AB - Contingent magnetic variation (CMV) data were recorded in three healthy male subjects using a 2 x 37 biomagnetometer system. The experiment was repeated for one of the subjects using a 151 whole-head biomagnetometer; the same auditory GO/NOGO choice reaction time paradigm as in the first experiment was used, extended to include repetitions of identical runs and additional control conditions. Magnetic field tomography was applied to the averaged data of each subject, for each run and condition (e.g., GO/NOGO). An independent estimate of the current density in the brain was obtained every few milliseconds. The slow components were emphasized by integrating the square of the current density vector, pixel by pixel, revealing in each subject activity in the auditory cortex, sensorimotor cortex, inferior prefrontal area, and posterior inferior parietal area. The intersubject variability was large, but looking across subjects the auditory and sensorimotor cortex (which were best covered by the two probes) were consistently identified in each subject as contributing to the generation of the early and late slow CMV components. These findings were confirmed by the whole-head single-subject experiment, in which slow activity was also identified in the supplementary motor area (SMA) and posterior cingulate cortex (PCC), areas very likely missed in the first experiment because of the limited view of the twin system. The PCC and particularly the SMA activations were substantially reduced when identical runs were repeated. PMID- 10694460 TI - The relationship between fMRI activation and cerebral atrophy: comparison of normal aging and alzheimer disease. AB - Functional MRI has recently been used to examine activation associated with aging and dementia, yet little is known regarding the effect of cerebral atrophy on fMRI signal. The purpose of this study was to examine the relationship between measures of global and regionally specific atrophy and fMRI activation in normal aging and in Alzheimer disease (AD). Two groups of subjects were studied with echoplanar imaging and quantitative structural volumetry: healthy controls spanning a broad age and atrophy range (n = 16) and patients with mild AD (n = 8). Results from a semantic task previously found to activate left inferior frontal (LIFG) and left superior temporal (LSTG) gyri were analyzed. The correlations between clusters of activation in the LIFG and LSTG and measures of local atrophy in the LIFG and LSTG regions were evaluated. For control subjects, there was no significant correlation between activation and regional or total brain atrophy (for LIFG r = -0.03, NS; for LSTG r = 0.20, NS). In contrast, for AD patients, there was a significant positive correlation between atrophy and activation in LIFG (r = 0.70, P = 0.05) but not LSTG (r = 0.00, NS). These results suggest that activation of language regions and atrophy within those regions may be independent among healthy adults spanning a broad age and atrophy range. However, in AD, a relationship exists in the LIFG that may reflect compensatory recruitment of cortical units or disease-specific changes in the hemodynamic response. PMID- 10694461 TI - fMRI of the responses to vibratory stimulation of digit tips. AB - Three studies were carried out to assess the applicability of fMRI at 3.0 T to analysis of vibrotaction in humans. A novel piezoelectric device provided clean sinusoidal stimulation at 80 Hz, which was initially applied in separate runs within a scanning session to digits 2 and 5 of the left hand in eight subjects, using a birdcage RF (volume) coil. Significant clusters of activation were found in the primary somatosensory cortex (SI), the secondary somatosensory cortex (SII), subcentral gyrus, the precentral gyrus, posterior insula, posterior parietal regions (area 5), and the posterior cingulate. Digit separation in SI was possible in all subjects and the activation sites reflected the known lateral position of the representation of digit 2 relative to that of digit 5. A second study carried out in six additional subjects using a surface coil, replicated the main contralateral activation patterns detected in study one and further improved the discrimination of the digits in SI. Significant digit separation was also found in SII and in the posterior insula. A third study to investigate the frequency dependence of the response focused on the effect of an increase in vibrotactile frequency from 30 to 80 Hz, with both frequencies applied to digit 2 during the same scanning session in four new subjects. A significant increase in the number of pixels activated within both SII and the posterior insula was found, while the number of pixels activated in SI declined. No significant change in signal intensity with frequencies was found in any of the activated areas. PMID- 10694462 TI - Dissociable temporal lobe activations during emotional episodic memory retrieval. AB - The richness of human recollective experience is, in part, related to evocation of previously experienced emotions. An extensive functional neuroimaging literature has provided a description of brain regions involved in retrieving emotionally neutral episodic memories. Whether similar or distinct systems are involved in retrieving emotional memories is unresolved. This question motivated the present functional neuroimaging study, using 0-15 positron emission tomography (PET), where we compared patterns of brain activation associated with retrieving previously studied emotional and neutral pictorial material. By varying task requirements and item density we characterized two distinct neural response patterns during emotional memory retrieval. First, we identified an anterior temporal pole activation that reflected the psychological set associated with emotional memory retrieval. Second, we identified a left amygdala response sensitive to actual retrieval of emotional items. These data suggest distinct functional roles for temporal lobe regions during emotional memory retrieval involving context-related tonic anterior temporal pole activation and a phasic item-related amygdala response. We conclude that brain regions involved in episodic memory retrieval reflect not only physical attributes of stimulus material, for example, their verbal or visual qualities, but also their affective significance. PMID- 10694463 TI - Covert visual spatial orienting and saccades: overlapping neural systems. AB - We used functional magnetic resonance imaging (fMRI) to investigate the functional anatomical relationship between covert orienting of visual spatial attention and execution of saccadic eye movements. Brain areas engaged by shifting spatial attention covertly and by moving the eyes repetitively toward visual targets were compared and contrasted directly within the same subjects. The two tasks activated highly overlapping neural systems and showed that common parietal and frontal regions are more activated during the covert task than the overt oculomotor condition. The possible nature of the relationship between these two operations is discussed. PMID- 10694464 TI - The role of the right anterior prefrontal cortex in episodic retrieval. AB - Regional brain activity was measured with H(2) (15)O PET while participants attempted to complete word-stem and word-fragment retrieval cues with previously studied words. The retrieval cue manipulation was employed to gain control over the monitoring operations associated with evaluating the episodic status of alternative cue completions. These operations were more constrained for fragments, which had fewer possible completions than each corresponding stem. In one condition (zero target), during the scanning interval none of the cues could be completed with studied items, whereas in another condition (high target), 80% of cues belonged to studied items. Relative to baseline tasks, right anterior prefrontal activity was greater for stems than for fragments in the zero target condition. The target density manipulation did not modulate right anterior prefrontal activity, but was associated with increased activity in right dorsolateral prefrontal cortex. These findings are consistent with the proposal that the right anterior prefrontal cortex supports monitoring operations during episodic retrieval tasks. In addition, the findings add to evidence suggesting that the dorsolateral and anterior right prefrontal cortex make functionally distinct contributions to episodic retrieval. PMID- 10694465 TI - Functional localization of a "Time Keeper" function separate from attentional resources and task strategy. AB - The functional neuroanatomy of time estimation has not been well-documented. This research investigated the fMRI measured brain response to an explicit, prospective time interval production (TIP) task. The study tested for the presence of brain activity reflecting a primary time keeper function, distinct from the brain systems involved either in conscious strategies to monitor time or attentional resource and other cognitive processes to accomplish the task. In the TIP task participants were given a time interval and asked to indicate when it elapsed. Two control tasks (counting forwards, backwards) were administered, in addition to a dual task format of the TIP task. Whole brain images were collected at 1.5 Tesla. Analyses (n = 6) yielded a statistical parametric map (SPM ?z?) reflecting time keeping and not strategy (counting, number manipulation) or attention resource utilization. Additional SPM ?z?s involving activation associated with the accuracy and magnitude the of time estimation response are presented. Results revealed lateral cerebellar and inferior temporal lobe activation were associated with primary time keeping. Behavioral data provided evidence that the procedures for the explicit time judgements did not occur automatically and utilized controlled processes. Activation sites associated with accuracy, magnitude, and the dual task provided indications of the other structures involved in time estimation that implemented task components related to controlled processing. The data are consistent with prior proposals that the cerebellum is a repository of codes for time processing, but also implicate temporal lobe structures for this type of time estimation task. PMID- 10694466 TI - Time-dependent changes in learning audiovisual associations: a single-trial fMRI study. AB - Functional imaging studies of learning and memory have primarily focused on stimulus material presented within a single modality (see review by Gabrieli, 1998, Annu. Rev. Psychol. 49: 87-115). In the present study we investigated mechanisms for learning material presented in visual and auditory modalities, using single-trial functional magnetic resonance imaging. We evaluated time dependent learning effects under two conditions involving presentation of consistent (repeatedly paired in the same combination) or inconsistent (items presented randomly paired) pairs. We also evaluated time-dependent changes for bimodal (auditory and visual) presentations relative to a condition in which auditory stimuli were repeatedly presented alone. Using a time by condition analysis to compare neural responses to consistent versus inconsistent audiovisual pairs, we found significant time-dependent learning effects in medial parietal and right dorsolateral prefrontal cortices. In contrast, time-dependent effects were seen in left angular gyrus, bilateral anterior cingulate gyrus, and occipital areas bilaterally. A comparison of paired (bimodal) versus unpaired (unimodal) conditions was associated with time-dependent changes in posterior hippocampal and superior frontal regions for both consistent and inconsistent pairs. The results provide evidence that associative learning for stimuli presented in different sensory modalities is supported by neural mechanisms similar to those described for other kinds of memory processes. The involvement of posterior hippocampus and superior frontal gyrus in bimodal learning for both consistent and inconsistent pairs supports a putative function for these regions in associative learning independent of sensory modality. PMID- 10694467 TI - SAP family proteins. AB - Thus far, five members including Dlg, SAP97/hDlg, SAP90/PSD-95, SAP102, and PSD 93/chapsyn110 which belong to SAP family have been identified. Recent studies have revealed that these proteins play important roles in the localization and function of glutamate receptors and K(+) channels. Although most of them have been reported to be localized to the synapse, only one member, SAP97, is expressed also in the epithelial cells. In this review, we have summarized structural characters of SAP family proteins and discuss their functions in neurons and epithelial cells. PMID- 10694468 TI - Molecular and immunochemical evidences demonstrate that endooligopeptidase A is the predominant cytosolic oligopeptidase of rabbit brain. AB - Oligopeptidases are tissue endopeptidases that do not attack proteins and are likely to be involved in the maturation and degradation of peptide hormones and neuropeptides. The rabbit brain endooligopeptidase A and the rat testes soluble metallopeptidase (EC 3.4.24.15) are thiol-activated oligopeptidases which are able to generate enkephalin from a number of opioid peptides and to inactivate bradykinin and neurotensin by hydrolyzing the same peptide bonds. A monospecific antibody raised against the purified rabbit brain endooligopeptidase A allowed the identification of a 2. 3 kb cDNA coding for a truncated enzyme of 512 amino acids, displaying the same enzymatic features as endooligopeptidase A. In spite of all efforts, employing several strategies, the full-length cDNA could not be cloned until now. The analysis of the deduced amino acid sequence showed no similarity to the rat testes metalloendopeptidase sequence, except for the presence of the typical metalloprotease consensus sequence [HEXXH]. The antibody raised against recombinant endooligopeptidase A specifically inhibited its own activity and reduced the thiol-activated oligopeptidase activity of rabbit brain cytosol to less than 30%. Analysis of the endooligopeptidase A tissue distribution indicated that this enzyme is mainly expressed in the CNS, whereas the soluble metallo EC 3.4.24.15 is mainly expressed in peripheral tissues. PMID- 10694469 TI - Characterization of Pen n 13, a major allergen from the mold Penicillium notatum. AB - Penicillium notatum is a well-known indoor aeroallergen and is frequently included in skin test panels for allergic diagnosis. On two-dimensional immunoblotting using patients' sera containing IgE and monoclonal antibody D7B8 specific for Pen c 1 of P. citrinum, two allergens with a molecular mass of 33 kDa but different isoelectric points were identified. A novel cDNA coding for Pen n 13 was cloned and sequenced. The nucleotide sequence codes for a protein 397 amino acids including a putative signal peptide of 25 amino acids and a propeptide of 90 amino acids. The allergen is an alkaline serine protease that shares more than 39% identical residues with other kinds of mold allergens. The coding cDNA of Pen n 13 was cloned into vector pQE-30 and expressed in E. coli M15 as a His-tag fusion protein and purified to homogeneity. The fusion protein reacted with monoclonal antibodies of Pen c 1 and with IgE from Penicillium allergic patients. Furthermore, it also cross-reacted strongly with IgE specific for the natural Pen c 1, indicating that similar IgE binding epitopes may exist in the allergens of P. notatum and P. citrinum. Antigenicity index plots indicated that there are several similar epitope regions of high antigenic indices in Pen c 1 and Pen n 13, corroborating that mold allergens belonging to the alkaline serine protease family possess similar protein structure and strong antigenic cross-reactivity. PMID- 10694470 TI - Markedly decreased binding of vincristine to tubulin in vinca alkaloid-resistant Chinese hamster cells is associated with selective overexpression of alpha and beta tubulin isoforms. AB - Vinca alkaloids are among a number of cytotoxic agents which target tumor cell microtubules. Studies described herein document the basis for one form of acquired resistance to these plant alkaloids involving an alteration of tubulin in a variant (DC-3F/VCRd-5L) of DC-3F Chinese hamster cells. Our results revealed a markedly decreased binding of [(3)H]vincristine (VCR) to tubulin extracted from this variant compared to tubulin extracted from wild-type DC-3F cells. This was quantitated as a 10- to 15-fold decrease in on-rate in the presence of GTP for the [(3)H]VCR associating with tubulin in cell-free cytosol and a 10-fold increase in off-rate for GTP-dependent dissociation of the [(3)H]VCR-tubulin complex. Quantitative RT-PCR and nucleotide sequencing of poly(A)(+) RNA also carried out with variant and wild-type DC-3F cells documented a different pattern of relative expression, but no base pair differences in the open reading frame of the three alpha and beta tubulin isoforms detected in each cell type. This was accounted for by selective overexpression of one alpha tubulin (alphaII) and two beta tubulin (betaI and betaIV) isoforms in the variant cells. These results would appear to provide an underlying basis for the large decrease in [(3)H]VCR binding by tubulin in these variant Chinese hamster cells and a major component of their acquired resistance to this vinca alkaloid. PMID- 10694471 TI - Digital fluorescence imaging of trafficking of endosomes containing low-density lipoprotein in brain astroglial cells. AB - We have used digital fluorescence microscopy to examine transport of LDL containing endosomes in rat brain astroglial cells to show that individual middle endosomes undergo rapid transitions between forward/backward movements and immobile states over short distances. The population of rapidly moving endosomes (>0.04 microm/sec) was 35. 9%, and the remaining endosomes were slowly moving or temporarily immobile (<0.04 microm/sec). The averaged motion was, however, a very slow perinuclear motion with a velocity of 3.25 microm/h. This small velocity is mainly due to frequent changing of directions in movements, requiring 6 h for a significant concentration around the circumference of the cell nuclei. The application of both anti-dynein antibodies and vanadate in permeabilized cells resulted in peripherally concentrated distribution of endosomes, probably due to inhibition of perinuclear motion by dynein-like motor proteins. These results imply that both dynein-like and kinesin-like proteins bind to the same endosome resulting in both perinuclear and peripherally directed movements. PMID- 10694472 TI - Activation of the human p27(Kip1) promoter by IFNalpha 2b. AB - p27(Kip1) is one of the key regulatory proteins in cell cycle through inhibition of pRB phosphorylation by suppression of the activity of several cyclin/Cdk complexes. The expression of p27(Kip1) has been shown to be controlled by a posttranslational mechanism, although vitamin D(3) and neuronal differentiation can also induce its mRNA. Recently, the p27(Kip1) promoter was isolated and sequenced from a human leukocyte genomic library. In this report, we demonstrate that IFNalpha 2b, activates the human p27(Kip1) promoter-driven luciferase reporter gene in transient expression assays in H82 cells. This induction might involve two IRF 1-like binding sites present in the p27(Kip1) promoter. To our knowledge this is the first report on the direct activation of the human p27(Kip1) promoter by IFNalpha 2b. PMID- 10694473 TI - BanI restriction endonuclease binds in the major groove of DNA: an inhibition kinetic study using substrates with mismatch basepairs. AB - Structural information on BanI-DNA interaction was obtained from simple inhibition kinetic assays using altered substrates. Self-complementary 13-mer oligodeoxynucleotides with or without mismatch basepairs in the BanI recognition sequence (GGPyPuCC) were synthesized. UV melting curves and CD spectra indicated double-stranded B-DNA structure for all the oligomers. Among the seven oligomers with recognition sequences, GGTACC, GGTGCC, GGTATC, GGCACC, GGAGCC, GGTAAC, and GGATCC, only the first two were cleaved with BanI. Kinetics of BanI cleavage of the native substrate was inhibited competitively by all of the other oligomers except the one with sequence GGCACC. From inhibition kinetic analysis in presence of a fixed concentration of the inhibitor, apparent K(m) and K(I) were determined. The data were analyzed in the context of alterations made in the hydrogen bonding potential in the major and minor groove of DNA within the recognition sequence due to basepair mismatches. Such analyses led to the conclusion that BanI, like BamHI, binds in the major groove and the central thymines make important contact with the protein. PMID- 10694474 TI - Leptin regulation of prepro-orexin and orexin receptor mRNA levels in the hypothalamus. AB - The aim of this study was to determine the effects of leptin treatment on prepro orexin and orexin receptor expression in the rat hypothalamus. Adult male rats, food-deprived for 48 and 72 h, were treated one time with vehicle or leptin (10 microg, icv). Prepro-orexin mRNA content was measured by semiquantitative RT-PCR, Northern blot, and in situ hybridization; orexin receptor 1 and 2 mRNA content was quantified by Northern blot and/or semiquantitative RT-PCR. Our results indicate that leptin inhibits a fasting-induced increase in prepro-orexin mRNA and orexin receptor 1 mRNA levels in the rat hypothalamus, while orexin receptor 2 mRNA levels were unchanged in all situations evaluated. These data provide direct evidence for an additional mechanism of adaptation of the hypothalamus to food deprivation and for a new effect of leptin in the regulation of food intake. PMID- 10694475 TI - Serum-free, long-term cultures of human hepatocytes: maintenance of cell morphology, transcription factors, and liver-specific functions. AB - Since human hepatocytes are available only in limited number, the development of a serum-free culture system for long-term cultivation of differentiated and functional hepatocytes is of great importance. Here we describe the culture of human hepatocytes in a chemically defined serum-free medium for up to 5 weeks. Cell morphology was assayed by light and electron microscopy and revealed a well preserved cellular morphology. Marker proteins for epithelial and bile duct cells, cytokeratin (CK) 18 and 19, and liver-specific proteins, like phosphoenolpyruvate carboxykinase-2 (PCK2) and serum proteins, were expressed. Liver-enriched transcription factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and hepatocyte nuclear factor-4 (HNF-4), cytokine and mitogen activated factors (nuclear factor kappa B) NFkappaB, and activator protein-1 (AP 1) were maintained and active for several weeks in our cultures. In summary, our serum-free culture system allows the culture of differentiated human hepatocytes for several weeks. It may serve as a model system for metabolic, pharmacologic toxicologic studies, and studies on human pathogens under defined chemical conditions. PMID- 10694476 TI - Cloning and characterization of the human retinoid X receptor alpha gene: conservation of structure with the mouse homolog. AB - Retinoid X receptors (RXRs) are members of the steroid/thyroid hormone receptor superfamily which, along with retinoic acid receptors (RARs), mediate the biological effects of retinoids. These effects include the regulation of many aspects of embryonic development, reproductive and visual function, and the maintenance of epithelial homeostasis throughout life. The genes for three distinct retinoid X receptors, RXRalpha, beta, and gamma, have been localized to separate chromosomes. In order to determine the organization of the human RXRalpha gene, we have isolated a clone containing the majority of the gene from a human genomic bacterial artificial chromosome (BAC) library and generated a physical map. The gene spans over 40 kilobases in size and contains at least 10 exons. Comparison with mapped portions of the mouse RXRalpha gene indicates highly conserved intron-exon positioning. These results provide information necessary to generate constructs for targeting the RXRalpha gene in human cell lines, which may eventually lead to an understanding of the function of RXRalpha in human cancer. PMID- 10694477 TI - Engineering His(E7) affects the control of heme reactivity in Aplysia limacina myoglobin. AB - Aplysia limacina myoglobin lacks the distal histidine (His (E7)) and displays a ligand stabilization mechanism based on Arg(E10). The double mutant Val(E7)His Arg(E10)Thr has been prepared to engineer the role of His(E7), typical of mammalian myoglobins, in a different globin framework. The 2.0 A crystal structure of Val(E7)His-Arg(E10)Thr met-Mb mutant reveals that the His(E7) side chain points out of the distal pocket, providing an explanation for the observed failure to stabilize the Fe(II) bound oxygen in the ferrous myoglobin. Moreover, spectroscopic analysis together with kinetic data on azide binding to met myoglobin are reported and discussed in terms of the presence of a water molecule at coordination distance from the heme iron. PMID- 10694478 TI - Characterization of mutations induced by ethyl methanesulfonate, UV, and trimethylpsoralen in the nematode Caenorhabditis elegans. AB - The genome project of the nematode Caenorhabditis elegans is completed. It is important and useful to disrupt nematode genes to know their function. We treated wild-type animals with potential candidates for mutagens for reverse genetics, EMS (ethyl methanesulfonate), short-wavelength UV, and long-wavelength UV in the presence of TMP (trimethylpsoralen). We estimated forward mutation rates by counting the occurrence of a marker unc-22 mutation. We found that the forward mutation rate by TMP/UV could be comparable with EMS by improving the frequency one order higher than before. We next isolated mutants of another marker gene ben 1 and examined the probability for the deletion mutations by PCR and sequencing. Deletion mutations were found only by TMP/UV method, which suggested TMP/UV is the choice for deletion mutagenesis among these methods. As a pilot experiment, we could isolate actual deletion mutations at a much higher frequency than previously. PMID- 10694479 TI - Regulation of rheumatoid synovial cell growth by ceramide. AB - Overgrowth of rheumatoid synoviocytes, which results in joint destruction, is due to impaired balance between cell proliferation and cell death (apoptosis). Ceramide is an important lipid messenger involved in mediating a variety of cell functions including apoptosis. We investigated the effects of ceramide on growth promoting anti-apoptotic signals in rheumatoid synovial cells. Human synovial cells isolated from patients with rheumatoid arthritis (RA) were stimulated with platelet-derived growth factor (PDGF) in the presence or absence of C2-ceramide. The kinase activity of Akt, MEK, and ERK1/2 was analyzed in PDGF-stimulated synovial cells by Western blot analysis. Pretreatment with C2-ceramide completely inhibited PDGF-induced cell cycle progression of rheumatoid synovial cells. PDGF stimulation induced phosphorylation and activation of Akt, MEK, and ERK1/2 in rheumatoid synovial cells. C2-ceramide inhibited the activation of Akt, MEK and ERK1/2 in PDGF-stimulated synovial cells. Our data demonstrated that inhibition of anti-apoptotic kinases, such as Akt and ERK1/2, may play an important role in ceramide-mediated apoptosis of rheumatoid synovial cells. PMID- 10694480 TI - Assignment of overlapping (1)H NMR signals in carp seminal plasma by proton detected 2D C,H correlation spectroscopy. AB - The (1)H NMR spectrum of the perchloric acid extract of carp seminal plasma was heavily congested. It is demonstrated that proton-detected C,H chemical shift correlation spectroscopy (HSQC, HSQC-TOCSY) allows an unequivocal identification of proline, glutamate, taurine, and methionine sulfoxide, although several key proton signals were strongly overlapped. PMID- 10694481 TI - Design, chemical synthesis and kinetic studies of trypsin chromogenic substrates based on the proteinase binding loop of Cucurbita maxima trypsin inhibitor (CMTI III). AB - A series of trypsin chromogenic substrates with formula: Y-Ala-X-Abu-Pro-Lys-pNA, where X = Gly, Ala, Abu, Val, Leu, Phe, Ser, Glu and Y = Ac, H; pNA = p nitroanilide was synthesized. The Cucurbita maxima trypsin inhibitor CMTI-III molecule was used as a vehicle to design the trypsin substrates. To evaluate the influence of position P(4) on the substrate-enzyme interaction, kinetic parameters of newly synthesized substrates with bovine beta-trypsin were determined. The increasing hydrophobicity of the amino acid residue (Gly, Ala, Abu, Val) introduced in position P(4) significantly enhanced the substrate specificity (k(cat)/K(m)) which was over 8 times higher for the last residue than that for the first one. The introduction of residues with more hydrophilic side chain (Glu, Ser) in this position reduced the value of this parameter. These results correspond well with those obtained using molecular dynamics of bovine beta-trypsin with monosubstituted CMTI-I analogues, indicating that in both trypsin substrate and inhibitor position 4 plays an important role in the interaction with the enzyme. PMID- 10694482 TI - Flow cytometric parameters for characterizing platelet activation by measuring P selectin (CD62) expression: theoretical consideration and evaluation in thrombin treated platelet populations. AB - Two flow cytometric parameters are generally used to quantify platelet activation as measured by P-selectin (CD62) expression: percentage and mean channel fluorescence of CD62-positive platelets (%(+) and MCF(+), respectively). We describe a method for calculation of indices of platelet activation for positive (IPA(+)) and total (IPA(Sigma)) platelets, which reflect integrated amounts of CD62 expressed in these populations; IPA(+) is calculated as the product of %(+) and MCF(+), whereas IPA(Sigma) is exclusively determined by mean fluorescence of the total platelet population (MCF(Sigma)) and does not depend on %(+). We use these parameters to characterize human platelet activation in whole blood samples treated with varying human alpha-thrombin concentrations, mimicking the variations in platelet activation in a number of clinical settings. Multiparameter analysis of CD62 expression may be useful for selective diagnosis of disorders with systemic or localized platelet activation and for monitoring the clinical course of the disease and effect of therapeutic interventions. PMID- 10694483 TI - Molecular cloning and genomic organization of a novel receptor from Drosophila melanogaster structurally related to mammalian galanin receptors. AB - We screened the Berkeley "Drosophila Genome Project" database with "electronic probes" corresponding to conserved amino acid sequences from the five known rat somatostatin receptors. This yielded alignment with a Drosophila genomic clone that contained a DNA sequence coding for a protein, having amino acid sequence identities with the rat galanin receptors. Using PCR with Drosophila cDNA as a template, and oligonucleotide probes coding for the exons of the presumed Drosophila gene, we were able to clone the cDNA for this receptor. The Drosophila receptor has most amino acid sequence identity with the three mammalian galanin receptors (37% identity with the rat galanin receptor type-1, 32% identity with type-2, and 29% identity with type-3). Less sequence identity exists with the mammalian opioid/nociceptin-orphanin FQ receptors (26% identity with the rat micro opioid receptor), and mammalian somatostatin receptors (25% identity with the rat somatostatin receptor type-2). The novel Drosophila receptor gene contains ten introns and eleven exons and is located at the distal end of the X chromosome. PMID- 10694484 TI - Cerivastatin suppresses lipopolysaccharide-induced ICAM-1 expression through inhibition of Rho GTPase in BAEC. AB - We investigated the effect of cerivastatin on lipopolysaccharide (LPS)-induced intercellular adhesion molecule-1 (ICAM-1) expression in bovine aortic endothelial cells. Cerivastatin suppressed LPS-induced ICAM-1 mRNA expression. Cotreatment with geranylgeranylpyrophosphate reversed the effect of cerivastatin. Because Rho undergoes geranylgeranyl modification, we elucidated whether Rho is involved in LPS-induced ICAM-1 expression. Inhibition of Rho activity by Clostridium botulinum C3 transferase or by overexpression of RhoA T19N, a dominant-negative mutant of RhoA, decreased LPS-induced ICAM-1 expression. Although cerivastatin up-regulated endothelial nitric oxide synthase (eNOS), inhibition of nitric oxide (NO) synthesis by cotreatment with N(omega)-nitro-l arginine methyl ester (L-NAME) exhibited no influence on the effect of cerivastatin. The present results indicate that cerivastatin prevents LPS-induced ICAM-1 expression in endothelial cells via inhibition of Rho activity. This inhibitory effect is likely unrelated to up-regulation of eNOS. PMID- 10694485 TI - Molecular cloning and genomic structure of the betaTRCP2 gene on chromosome 5q35.1. AB - Beta-catenin, IkappaBalpha, and HIV Vpu are recruited to the ubiquitin-proteasome degradation pathway by betaTRCP, one of the components of the ubiquitin ligase complex. betaTRCP2, a related gene of betaTRCP, was cloned and characterized. Three isoforms, betaTRCP2A, betaTRCP2B, and betaTRCP2C, were identified. All of these betaTRCP2 isoforms consist of an F-box and seven WD repeats. Human betaTRCP2A shows 86% total amino acid identity with human betaTRCP. betaTRCP2 mRNA of 4.5 kb in size was detected almost ubiquitously. Sequence analyses on betaTRCP2 genomic clones revealed that the betaTRCP2 gene consists of at least 14 exons. Exons 1 and 4-14 are shared among all betaTRCP2 isoforms. betaTRCP2A of 508 amino acids lacks exons 2 and 3, betaTRCP2B of 529 amino acids contains exon 3, and betaTRCP2C of 542 amino acids contains exon 2. These results indicate that three betaTRCP2 isoforms are transcribed due to alternative splicing. The betaTRCP2 gene has been mapped to human chromosome 5q35.1 by fluorescence in situ hybridization. PMID- 10694486 TI - Comprehensive gene expression profile of a normal human liver. AB - To investigate the gene expression profile of a normal human liver, we performed serial analysis of gene expression (SAGE), which allows the quantitative and simultaneous analysis of thousands of genes expressed in tissue. Polyadenylated RNA was obtained from a bulk normal human liver sample and SAGE was performed. Reverse transcriptase-polymerase chain reaction (RT-PCR) was also performed in each of 3 different normal liver samples to evaluate the validity of the profile in each individual. A total of 30,982 tags were sequenced, 8,596 of which were unique. The genes highly expressed in the normal liver were those encoding plasma proteins (>21.8% of total transcripts), cytoplasmic proteins (>8.6%), enzymes (>4.8%), protease inhibitors (>1.7%), complements (>1.1%), and coagulation factors (>0.75%). About 13.9% of all transcripts encoded genes not reported in GenBank thus far. This study identifies candidate genes to be examined in relation to various human liver diseases, including viral hepatitis, liver cirrhosis, and hepatocellular carcinoma. PMID- 10694487 TI - Chemotaxis of a Ralstonia sp. SJ98 toward different nitroaromatic compounds and their degradation. AB - A Ralstonia sp. SJ98, isolated by a chemotactic enrichment technique, was capable of utilizing different nitroaromatic compounds (NACs). It utilized p-nitrophenol, 4-nitrocatechol, o-nitrobenzoic acid, and p-nitrobenzoic acid as the sole source of carbon and energy. It was observed that Ralstonia sp. SJ98 was chemotactic to the above-mentioned NACs as tested by the drop assay, swarm plate assay, and capillary assay. However, it failed to show chemotactic behavior toward those compounds which were not degraded by the microorganism. This is the first report which shows the chemotaxis of a microorganism toward different NACs and their subsequent degradation. Some of the intermediates of the NACs' degradative pathways have been identified using TLC, GC, and GC-MS studies. The results presented here indicate a correlation between chemotaxis and biodegradation of NACs. PMID- 10694488 TI - Structural basis for SH2D1A mutations in X-linked lymphoproliferative disease. AB - X-linked lymphoproliferative disease (XLP) is a rare and severe immune deficiency, characterized by abnormal immune responses to the Epstein-Barr virus. Recently, the gene responsible for XLP, SH2D1A, has been identified and shown to code for a small cytoplasmic protein with an SH2 domain that interacts with SLAM and 2B4, two receptorial molecules involved in signal transduction in T and NK cells, respectively. A variety of SH2D1A gene mutations have been reported thus far in XLP males. Here we describe a single-strand conformation polymorphism assay for mutation analysis in XLP. Four novel patients with SH2D1A mutations are described. These mutants, and the others previously reported in the literature, have been included in a Registry (SH2D1Abase) that is fully accessible on the World Wide Web. A three-dimensional model of the SH2 domain of the SH2D1A protein has been developed, based on homology with other SH2 domains. The structural consequences of disease-causing SH2D1A mutations are discussed. PMID- 10694489 TI - Chemiluminescent detection and imaging of reactive oxygen species in live mouse skin exposed to UVA. AB - The recent increase of ultraviolet (UV) rays on Earth due to the increasing size of the ozone hole is suggested to be harmful to life and to accelerate premature photoaging of the skin. The detrimental effects of UV radiation on the skin are associated with the generation of reactive oxygen species (ROS) such as superoxide anion radical (*O(-)(2)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (*OH), and singlet oxygen ((1)O(2)). However, direct proof of such ROS produced in the skin under UV irradiation has been elusive. In this study, we report first in vivo detection and imaging of the generated ROS in the skin of live mice following UVA irradiation, in which both a sensitive and specific chemiluminescence probe (CLA) and an ultralow-light-imaging apparatus with a CCD camera were used. In addition, we found that *O(-)(2) is formed spontaneously and (1)O(2) is generated in the UVA-irradiated skin. This method should be useful not only for noninvasive investigation of the spatial distribution and quantitative determination of ROS in the skin of live animals, but also for in vivo evaluation of the protective ability of free radical scavengers and antioxidants. PMID- 10694490 TI - Translocation of HSP27 to sarcomere induced by ischemic preconditioning in isolated rat hearts. AB - We investigated the role of the 27-kDa heat shock protein (HSP27) in cardiac protection using Langendorff-perfused rat hearts. After preconditioning (a single episode of 5 min global ischemia followed by 5 min of reperfusion), HSP27 redistributed from the cytosol to the sarcomere and recovery of the contractile function, after 40 min of global ischemia and 50 min of reperfusion, was significantly enhanced. Both SB203580, a p38 MAP kinase inhibitor, and bisindolylmaleimide I, a protein kinase C inhibitor, prevented the effects of preconditioning. Both 2-chloro-N(6)-cyclopentyladenosine (adenosine A1 agonist) and anisomycin (activator of p38 MAP kinase and c-jun N-terminal kinase) mimicked preconditioning. These results suggest that activation of protein kinase C followed by activation of p38 MAP kinase elicits translocation of HSP27 to the sarcomere, a process which may be involved in the cardioprotective mechanism afforded by ischemic preconditioning in rat heart. PMID- 10694491 TI - Aminopeptidase N/CD13 is associated with raft membrane microdomains in monocytes. AB - Ectopeptidases play important roles in cell activation, proliferation, and communication. Human monocytic cells express considerable amounts of aminopeptidase N/CD13, a transmembrane protein previously proposed to play a role in the regulation of neuropeptides and chemotactic mediators as well as in adhesion and cell-cell interactions. Here, we report for the first time that aminopeptidase N/CD13 in monocytes is partially localized in detergent-insoluble membrane microdomains enriched in cholesterol, glycolipids, and glycosylphosphoinositol-anchored proteins, referred to as "rafts." Raft fractions of monocytes were characterized by the presence of GM1 ganglioside as raft marker molecule and by the high level of tyrosine-phosphorylated proteins. Furthermore, similar to polarized cells, rafts in monocytic cells lack Na(+), K(+)-ATPase. Cholesterol depletion of monocytes by methyl-beta-cyclodextrin greatly reduces raft localization of aminopeptidase N/CD13 without affecting ala-p-nitroanilide cleaving activity of cells. PMID- 10694492 TI - Cre-mediated cerebellum- and hippocampus-restricted gene mutation in mouse brain. AB - Using the phage P1-derived Cre/loxP recombination system, we have created a line of cre-transgenic mice in which the Cre-mediated gene deletion is restricted to granule cells of cerebellum and dentate gyrus of hippocampus. Low levels of deletion were also present in pyramidal cells of hippocampal CA1 and CA3 fields. The Cre/loxP recombination occurred prenatally. The recombination efficiencies in the granular layer of the cerebellum, the granular layer of the dentate gyrus, and the CA1 and CA3 pyramidal cells of the hippocampus were 34.0%, 23.1%, 3.0%, and 9.8%, respectively. This line of cre-transgenic mice should be conducive to studies of the effect of a gene mutation upon brain development and plasticity. PMID- 10694493 TI - Dolichin, a new chitinase-like antifungal protein isolated from field beans (Dolichos lablab). AB - An antifungal protein, possessing a molecular weight of 28 kDa and an N-terminal sequence resembling chitinases, has been purified from the seeds of the field bean Dolichos lablab. The procedure involved extraction with aqueous buffer, affinity chromatography on Affi-gel blue gel, and ion exchange chromatography on CM-Sepharose. The protein, designated dolichin, exhibited antifungal activity against the fungi Fusarium oxysporum, Rhizoctonia solani, and Coprinus comatus. Dolichin was capable of inhibiting human immunodeficiency virus (HIV) reverse transcriptase and alpha- and beta-glucosidases which are glycohydrolases implicated in HIV infection. It had very low ribonuclease and cell-free translation-inhibitory activities. PMID- 10694494 TI - Attenuated nitric oxide synthase activity and protein expression accompany intestinal ischemia/reperfusion injury in rats. AB - Intestinal ischemia/reperfusion (I/R) leads to bowel impairment via the release of reactive oxygen species (ROS) and neutrophil infiltration. In addition to modulating intestinal integrity, nitric oxide (NO(*)) inhibits neutrophil activation and scavenges ROS. Attenuated endogenous NO(*) formation may result in the accrual of these deleterious stimuli. Therefore, we determined nitric oxide synthase (NOS) activity in anesthetized rats subjected to 1 h of superior mesenteric ischemia or ischemia followed by reflow. NOS activity was measured in intestinal tissue homogenates as the conversion rate of (3)H-L-arginine to (3)H-L citrulline. Our results demonstrate that intestinal ischemia leads to a decrease in NOS activity indicating lower NO(*) formation in the animal model. The attenuation in NOS activity was not reversed following 4 h of reperfusion. Western blot analysis revealed that the decline in enzyme activity was accompanied by reduced intestinal NOS III (endothelial constitutive NOS) expression. These findings provide biochemical evidence for impaired NO(*) formation machinery in intestinal I/R injury. PMID- 10694495 TI - Nitric-oxide-dependent pial arteriolar dilation in the female rat: effects of chronic estrogen depletion and repletion. AB - In this study, we compared endothelial nitric oxide synthase (eNOS)-mediated cerebral vasodilating responses in intact female rats, chronically ovariectomized (OVX) rats, and OVX rats treated for 2 weeks with 17beta-estradiol (E(2)). Under anesthesia, using intravital microscopy and a closed cranial window system, pial arteriolar diameter changes were monitored during sequential cortical suffusions of an eNOS-dependent dilator [acetylcholine (ACh)] and a direct NO donor [S nitrosoacetylpenicillamine (SNAP)]. In separate rats from the same groups, we compared eNOS and caveolin-1 (CAV-1) protein abundance in pial arterioles (via immunofluorescence analyses). In untreated and low-dose E(2)-treated (1.0 microg x kg(-1) x day(-1)) OVX rats, ACh-induced vasodilations were virtually absent. High-dose E(2) treatment (100 microg x kg(-1) x day(-1)) restored ACh-induced pial arteriolar dilations to levels seen in intact females. The vasodilations elicited by SNAP and ADO were unaffected by chronic estrogen changes, indicating no direct estrogen influence on vascular smooth muscle (VSM) reactivity. Pial arteriolar eNOS protein abundance was diminished by ovariectomy and restored by high-dose E(2) treatment. Pial arteriolar CAV-1 expression was higher in OVX versus intact and E(2)-treated OVX females. These results suggest that long-term changes in estrogen directly influence brain eNOS functional activity. The estrogen-related changes in eNOS-dependent vasodilating function appear to be related, in part, to a capacity for E(2) to increase eNOS protein expression and, in part, to an E(2)-associated diminution in endothelial CAV-1 expression. PMID- 10694496 TI - Microvascular pericytes express aggrecan message which is regulated by BMP-2. AB - Multipotential mesenchymal stem cells capable of chondro-osseous induction contribute to the endochondral callus of healing fractured bone. Microvascular pericytes serving the role of multipotential mesenchymal stem cells are considered osteoprogenitors because they express type I collagen, alkaline phosphatase enzyme activity, osteocalcin immunoreactivity, and bone sialoprotein mRNA. Previous electron microscopic studies indicate that this cell type has a contribution to the fracture callus. Limited data suggest that pericytes may also assume a chondrogenic phenotype. We undertook in vitro studies to understand how the chondro-osseous phenotype of the pericyte might be regulated. Using Northern analysis and semiquantitative reverse transcriptase-polymerase chain reaction (RT PCR), we found that cultured pericytes produce aggrecan and type II collagen mRNA indicating their chondrogenic potential. Aggrecan message is elevated by BMP-2 as analyzed by both Northern hybridization and RT-PCR. This finding suggests a regulatory role for this morphogen on this phenotype in pericytes. RT-PCR amplified versican product was also associated with pericyte cultures but was not affected by BMP-2. Our data strongly support a chondrogenic role for the pericyte and that the phenotype is regulated at least in part by BMP. PMID- 10694497 TI - p21(WAF1/CIP1) inhibits initiator caspase cleavage by TRAIL death receptor DR4. AB - Death receptors of the Tumor Necrosis Factor (TNF) family form membrane-bound self-activating signaling complexes that initiate apoptosis through cleavage of proximal caspases including CASP8 and 10. Here we show that overexpression of the cytoplasmic domain (CD) of the DR4 TRAIL receptor (TNFRSF10A, TRAIL R1) in human breast, lung, and colon cancer cell lines, using an adenovirus vector (Ad-DR4 CD), leads to p53-independent apoptotic cell death involving cleavage of CASP8 and 10 proximally and CASP3, 6, and 7 distally. DR4-CD overexpression also leads to cleavage of poly(ADP-ribose) polymerase (PARP) and the DNA fragmentation factor (DFF45; ICAD). Importantly, normal lung fibroblasts are resistant to DR4 CD overexpression and show no evidence of PARP-, CASP8- or CASP3-cleavage despite similar levels of adenovirus-delivered DR4-CD protein as the cancer cells. These results suggest that DR4 may signal death through known caspases and that further studies are required to evaluate Ad-DR4-CD as a novel anti-cancer agent. Finally, we show that overexpression of the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) (CDKN1A), or its N-terminal 91 amino acids containing cell cycle inhibitory activity, inhibits DR4-CD-dependent proximal caspase cleavage. The blockage of initiator caspase activation provides a novel insight into how p21 may suppress apoptosis and enhance cell survival. PMID- 10694498 TI - Localization of HSP90 binding sites in the human hepatitis B virus polymerase. AB - The fact that HSP90 proteins and their chaperonin partners play an important role in epsilon RNA binding of duck HBV Pol protein during duck HBV replication has been reported. To elucidate the molecular basis of HBV Pol/HSP90 interaction, we have characterized the HSP90 interaction to HBV Pol. We found that human HBV Pol protein upon synthesis in rabbit reticulocyte lysate formed a complex with HSP90 in vitro as duck HBV Pol did. In addition, HSP90 protein was copurified with MBP/POL protein expressed in HepG2 cells, suggesting that human HBV Pol protein is associated with HSP90 in vivo. To localize the HSP90 interaction site region, several deletion mutants of HBV Pol translated in vitro were immunoprecipitated with anti-HSP90 antibody. The result indicates that C-terminal regions of the TP and RT domains interact with HSP90 independently. PMID- 10694499 TI - Identification of a novel retinoic acid-responsive element within the lamin A/C promoter. AB - A-type lamins are not present in either early embryos or the embryonal carcinoma (EC) cell line. P19 cells, which are EC cell line, are able to express A-type lamins upon retinoic acid (RA) treatment. Here we report that a novel RA responsive element, termed lamin A/C-RA-responsive element (L-RARE), is located within the lamin A/C promoter. RA activated the luciferase activity of the reporter which had four tandem repeats of the wild-type L-RARE, while a loss of function mutant, which altered CACCCCC to CACtatC within L-RARE, did not respond. Four specific binding complexes of L-RARE, Complexes-A, -B, -C, and -D, were detected in protein extracts obtained from P19 cells treated with and without RA. Specific antibodies revealed that Sp1 and Sp3 were included in Complex-A and Complexes-B and -C, respectively. Thus, L-RARE was important in the RA-mediated activation of the lamin A/C promoter and was recognized by DNA binding proteins. PMID- 10694500 TI - Quinqueginsin, a novel protein with anti-human immunodeficiency virus, antifungal, ribonuclease and cell-free translation-inhibitory activities from American ginseng roots. AB - A homodimeric protein designated quinqueginsin, with a molecular weight of 53 kDa, has been isolated from the roots of American ginseng Panax quinquefolium. It was unadsorbed on DEAE cellulose in low ionic strength and neutral pH, and adsorbed on Affigel blue gel and SP-Sepharose under similar conditions. Its N terminal sequence bore similarity to those of plant ribosome inactivating proteins and fungal ribonucleases. The protein displayed a variety of biological activities. It possessed ribonucleolytic activity toward yeast tRNA and specific activity toward poly C. It inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC(50) of 0.26 nM, and exerted antifungal action against Fusarium oxysporum, Rhizoctonia solani, and Coprinus comatus. An inhibitory action was expressed toward human immunodeficiency virus-1 reverse transcriptase. This action was potentiated after chemical modification with succinic anhydride. PMID- 10694501 TI - Progestins and androgens increase expression of Spot 14 in T47-D breast tumor cells. AB - Enhanced expression of fatty acid synthase and other lipogenic enzymes has been observed in a subset of breast cancers with poor prognosis. This phenomenon has been related to amplification of a gene on chromosome region 11q13 encoding Spot 14, a putative regulator of lipogenic enzyme expression. In this paper we demonstrate that the induction of lipogenesis by progestins and androgens in the breast cancer cell line T47-D is accompanied by a marked increase in the expression of Spot 14. These data corroborate the correlation between Spot 14 expression and increased lipogenesis. Moreover they show that apart from gene amplification there is another steroid-regulated pathway that may enhance Spot 14 expression and lipogenesis in tumor cells. PMID- 10694502 TI - Molecular mechanism of the nacreous layer formation in Pinctada maxima. AB - We have cloned the cDNAs that encode two kinds of molluscan shell matrix proteins, namely N66 and N14, in the nacreous layer of Pinctada maxima. N66 is composed of carbonic anhydrase-like and repeat domains, as described for nacrein (1) in the pearls of P. fucata. N14 is homologous to N16, recently found in the nacreous layer of P. fucata (2) and is characterized by high proportions of Gly, Tyr, and Asn together with NG repeat sequences. The molecular weights of these proteins were estimated as 59,814 and 13,734 Da, respectively. Structural differences were clearly indicated in the alignment and length of the repeat sequences of the sets of the homogeneous proteins (N66/nacrein and N14/N16). The longer repeat sequences of N66 and N14 may be responsible for P. maxima's excellent property of calcification. The in vitro crystallization experiments revealed that the mixture of N66 and N14 could induce platy aragonite layers highly similar to the nacreous layer, once adsorbed onto the membrane of the water-insoluble matrix. PMID- 10694503 TI - Suppressive effects of swainsonine and N-butyldeoxynojirimycin on human bone marrow neutrophil maturation. AB - The effects of the N-linked oligosaccharide inhibitors swainsonine and N butyldeoxynojirimycin (NB-DNJ) on granulopoiesis was investigated using human bone marrow cells in in vitro liquid and agar cultures. The addition of the inhibitors into cultures containing granulocyte colony-stimulating factor (G-CSF) suppressed maturation from myelocytes into mature neutrophils. Swainsonine did not induce apoptosis, but NB-DNJ induced considerable apoptosis, especially in the presence of G-CSF. This result indicated that the decrease of mature neutrophils by swainsonine was not because of cell degeneration. In the case of NB-DNJ, it was thought to be because of both maturation suppression and apoptosis. In a colony-forming unit-granuloid (CFU-G) colony assay, the number of colonies was increased in the presence of the inhibitors, but the morphology of colonies was predominantly compact, or immature. The inhibitors also suppressed the expressions of mRNAs of CCAAT/enhancer binding protein epsilon (C/EBPepsilon) and G-CSF receptor as markers of terminal neutrophil maturation. These findings suggested that the incompleteness of N-linked oligosaccharide leads to the suppression of terminal neutrophil maturation. PMID- 10694504 TI - Enhanced prenyltransferase activity and Rab content in rat liver regeneration. AB - Rabs are small GTP-binding proteins with a regulatory role in intracellular vesicular traffic. The modulation of their levels and activity in different physiological situations is poorly understood. During the first cell cycle of rat liver regeneration we observed a differential regulation of some Rabs, with a progressive increase of those involved in exocytosis and a progressive decrease of one involved in endocytosis. This could be related with the need of exposing growth factor receptors and prolonging the transduction of their signal in preparation for mitosis. Moreover, we observed an increased activity of protein prenyltransferases, the enzymes responsible for the prenylation of several proteins involved in crucial processes of proliferation, without a corresponding increase in the amount of prenyltransferase protein. PMID- 10694506 TI - Structure-function relationships of the two surface loops of myosin heavy chain isoforms from thermally acclimated carp. AB - The structure-function relationships of fast skeletal myosin isoforms remain poorly understood. To shed some light, we constructed chimeric myosins comprised of Dictyostelium myosin heavy chain backbone with carp loop sequences and analyzed their functional properties. A loop 2-10 chimeric myosin having the loop 2 sequence of the fast skeletal isoform predominantly expressed in carp acclimated to 10 degrees C showed V(max) in actin-activated Mg(2+)-ATPase activity 1.4-fold higher than a loop 2-30 chimera constructed from the loop 2 sequence of the dominant isoform in carp acclimated to 30 degrees C. These two chimera exhibited no significant differences in sliding velocity of actin filaments in in vitro motility assay. Contrastingly, both loop 1-associated chimeras, loop 1-10 and loop 1-30, did not differ in both ATPase activity and in sliding velocity of actin filaments. PMID- 10694505 TI - Paraoxonase activity is reduced by a pro-atherosclerotic diet in rabbits. AB - Serum paraoxonase (PON1) is believed to protect against the development of atherosclerosis because of its ability to retard the oxidation of low-density lipoprotein (LDL) by hydrolysing LDL-associated phospholipid and cholesteryl ester hydroperoxides. We have examined the relationship between PON1 and atherosclerosis development in transgenic rabbits overexpressing human apolipoprotein (apo) A-I and nontransgenic littermates fed a pro-atherogenic diet. PON1 activity was higher in transgenic (4006.1 +/- 716.7 nmol/min/ml) compared to control (3078.5 +/- 623.3 nmol/min/ml) rabbits (P < 0.01) while high density lipoprotein (HDL) cholesterol was 1.84 +/- 0.54 mmol/L in transgenic rabbits and 0.57 +/- 0.21 mmol/L in control rabbits (P = 0.0001). After feeding rabbits a high-cholesterol diet for 14 weeks HDL-cholesterol fell by 70% in both transgenic and control rabbits (P < 0.001 compared to week 0) PON1 activity fell by 50% in both groups of rabbits (P < 0. 01 compared to week 0). The amount of thoracic aortic surface area covered by lesions was 29 +/- 16% in the control group and 26 +/- 15% in the transgenic group (P = NS). A pro-atherosclerotic diet reduces PON1 which may exaggerate the effects of the diet on the development of atherosclerosis. PMID- 10694507 TI - The cyclic AMP-protein kinase A pathway restrains islet phospholipase A(2) activation. AB - Although phospholipase A(2) (PLA(2)) is of importance for insulin secretion, it is not established how it relates to other signalling mechanisms. This study examined the crosstalk between PLA(2) and the cyclic AMP (cAMP)-protein kinase A (PKA) pathway in isolated rat islets. Forskolin, IBMX, and dbcAMP reduced [(3)H]arachidonic acid ([(3)H]AA) efflux from prelabelled islets during PLA(2) activation by mellitin or cholecystokinin (CCK-8), while efflux induced by carbachol was unaffected. The PKA inhibitor myrPKI(14-22) prevented this reduction of CCK-8-induced efflux. Glucagon-like peptide-1 (GLP-1), gastric inhibitory polypeptide (GIP), and vasoactive intestinal polypeptide (VIP) diminished CCK-8-induced efflux. Also in the absence of Ca(2+), forskolin/IBMX and dbcAMP reduced CCK-8-induced efflux. In parallel with effects on [(3)H]AA, the expected additive insulin secretion induced by mellitin or CCK-8 in combination with forskolin or GLP-1, respectively, was reduced. In conclusion, the cAMP-PKA pathway restrains both Ca(2+)-dependent and Ca(2+)-independent PLA(2) activation, indicating a regulating crosstalk between these two pathways. PMID- 10694508 TI - Helicobacter pylori induces formation of stress fibers and membrane ruffles in AGS cells by rac activation. AB - Helicobacter pylori induces signaling cascades leading to changes in cytoskeleton and an inflammatory response. Information on the morphological changes and cytoskeletal rearrangements induced by attachment of the bacterium is contradictory and signal transduction pathways are not well known. Since rho family of small GTPases is known to mediate cytoskeletal response to various extracellular stimuli, and is also involved in several other important signal transduction pathways, we have investigated the role of rac and cdc42 in H. pylori-induced cytoskeletal changes in cultured carcinoma AGS cells. AGS cells grown with serum expressed actin filaments in the form of short stress fibers and thin network at the edges, which were depolymerized by removal of serum. In serum starved cells both type I and type II strains of H. pylori induced formation of actin filaments and lamellipodia-like structures. Microinjection of active rac induced similar changes, but injection of inactive rac prevented the effects of H. pylori, while active or inactive cdc42 did not have any significant effect. Cytoskeletal effects of H. pylori were inhibited by actinomycin D, but not completely by cycloheximide. These results indicate that rac activation is involved in signal transduction cascade leading to cytoskeletal reorganization induced by H. pylori and that gene activation and synthesis of new proteins is necessary in this process. PMID- 10694509 TI - Binding of six chimeric analogs of omega-conotoxin MVIIA and MVIIC to N- and P/Q type calcium channels. AB - Replacement of the N-terminal half of omega-conotoxin MVIIC, a peptide blocker of P/Q-type calcium channels, with that of omega-conotoxin MVIIA significantly increased the affinity for N-type calcium channels. To identify the residues essential for subtype selectivity, we examined single reverse mutations from MVIIA-type to MVIIC-type in this chimeric analog. A reverse mutation from Lys(7) to Pro(7) decreased the affinity for both P/Q- and N-type channels, whereas that from Leu(11) to Thr(11) increased the affinity for P/Q-type channels and decreased the affinity for N-type channels. The roles of these two residues were confirmed by synthesizing two MVIIC analogs in which Pro(7) and Thr(11) were replaced with Lys(7) and Leu(11), respectively. PMID- 10694510 TI - Effects of shear stress on eicosanoid gene expression and metabolite production in vascular endothelium as studied in a novel biomechanical perfusion model. AB - This study investigated the effects of shear stress on gene expression of prostacyclin synthesis-related enzymes cyclooxygenases (COX-1 and COX-2), prostacyclin synthase (PGS), and thromboxane synthase (TXS) and their metabolites prostaglandin (PGI(2)) and thromboxane A(2) (TXA(2)) in endothelium of intact conduit vessels. Paired human umbilical veins were perfused at high/low shear stress (25/<4 dyn/cm(2)) at identical intraluminal pressure (20 mmHg) for 1.5, 3, or 6 hours in a computerized vascular model. High shear perfusion induced a significant, monophasic upregulation of PGS and TXS gene expressions after 6 hours. COX-1 and COX-2 mRNA showed a biphasic response with peaks at 1.5 and 6 hours, with a nadir level at 3 hours. Shear-induced gene expression was associated with a significantly greater accumulation of 6-keto prostaglandin F(1alpha) and TXA(2) in the perfusion medium. Thus, shear stress independently of perfusion pressure alters the expression of prostacyclin synthesis-related enzymes and the biosynthesis of their vasoactive metabolites. PMID- 10694511 TI - yam8(+), a Schizosaccharomyces pombe gene, is a potential homologue of the Saccharomyces cerevisiae MID1 gene encoding a stretch-activated Ca(2+)-permeable channel. AB - The Saccharomyces cerevisiae MID1 gene encodes a stretch-activated Ca(2+) permeable channel. In a protein database, we found a Schizosaccharomyces pombe gene whose predicted protein shows 26% identical and 62% similar to the Mid1 channel in amino acid sequence. cDNA derived from this gene, designated yam8(+), was isolated by reverse transcription-polymerase chain reaction (RT-PCR). Further analysis showed that the Yam8 protein consists of 486 amino acids and has 6 hydrophobic segments. The yam8(+) cDNA, placed under the S. cerevisiae TDH3 promoter, partially complemented the mating pheromone-induced death (mid) phenotype of the S. cerevisiae mid1 mutant. The expression of the yam8(+) cDNA in the mid1 mutant cells partially remediated the mid phenotype and resulted in a slight increase in Ca(2+) uptake activity. These findings suggest that Yam8 is a potential homologue of Mid1. PMID- 10694512 TI - Involvement of hepatic nuclear factor I binding motif in transcriptional regulation of Ca(2+)-binding protein regucalcin gene. AB - The characterization of the binding of nuclear protein on the 5'-flanking region of the rat regucalcin gene was investigated. Nuclear extracts from rat liver and H4-II-E hepatoma cells were used for oligonucleotide competition gel mobility shift assay. An oligonucleotide between position -523 and -506 in the 5'-flanking region of the rat regucalcin gene, which contains a nuclear factor I (NF1) consensus motif TTGGC(N)(6)CC, competed with the probe for the binding of the nuclear proteins from rat liver and H4-II-E cells. The mutation of TTGGC in the consensus sequence caused an inhibition of the binding of nuclear factors. The presence of Bay K 8644, insulin, and phorbol esters could stimulate the binding of the nuclear factors to the TTGGC region of the rat regucalcin gene in H4-II-E cells. The specific mutation introduced in this region, which was ligated to a luciferase reporter gene, reduced significantly the effects of Bay K 8644, insulin, and phorbol esters in stimulating the regucalcin gene transcriptional activity in H4-II-E cells. These results suggest that the specific nuclear factor binds to the NF1-like sequence, which can stimulate the transcriptional activity, in the promoter region of regucalcin gene in liver cells. PMID- 10694513 TI - Tyrosine phosphatase SHP-2 binding to CTLA-4: absence of direct YVKM/YFIP motif recognition. AB - CTLA-4 is well documented in its negative regulation of T-cell proliferation. However, little is known regarding the signaling mechanisms induced by CTLA-4. CTLA-4 associates with the phosphatidylinositol 3-kinase, the phosphatase SHP-2 and the clathrin adaptor complexes AP-1 and AP-2. SHP-2 SH2 domain binding to CTLA-4 is unusual given the absence of a I/VxYxxI/V/L motif. Here, we demonstrate that the phosphorylation of CTLA-4 tyrosines (YVKM and YFIP) fails to allow for single or tandem SHP-2 SH2 domain binding. This was observed using wild-type and inactive SHP-2 as well as a construct with the isolated two SH2 domains. The phosphorylated YVKM and YFIP motifs therefore do not appear to represent novel binding motifs for SHP-2 SH2 domains. At the same time, we could confirm that SHP 2 can associate with CTLA-4 in murine T-cells indicating that the interaction between the phosphatase and CTLA-4 is an indirect event, possibly mediated by PI 3-kinase/SHP-2 binding. PMID- 10694514 TI - Induction of leukemia cell differentiation and apoptosis by recombinant P48, a modulin derived from Mycoplasma fermentans. AB - P48 is a 48-kDa monocytic differentiation/activation factor which was originally identified in the conditioned medium of the Reh and other leukemia cell lines and has recently been shown to be a Mycoplasma fermentans gene product. Previously, conditioned medium P48 has been shown to induce differentiation of HL-60 (human promyelocytic leukemia) cells. Recently our laboratory isolated cDNA clones for P48 from Reh cells and genomic clones from Mycoplasma fermentans and expressed the recombinant protein as a maltose binding protein (MBP) fusion protein in E. coli. In this report we present the initial characterization of this recombinant P48 fusion protein (rP48-MBP). We show that rP48-MBP induces differentiation of HL-60, U937 (human histiocytic lymphoma), and M1 (mouse myeloid leukemia) cell lines. Interestingly, rP48-MBP also induces apoptosis of U937 and HL-60 cells as assessed by terminal transferase (TUNEL) assays. This is the first report of induction of apoptosis by a Mycoplasma gene product. P48 is a Mycoplasma-derived immunomodulatory molecule which has differentiation and apoptosis-inducing activities and may be important in the pathophysiology of Mycoplasma infections. The recombinant protein may be useful in studying the mechanisms of differentiation, cytokine production, and apoptosis in malignant and nonmalignant hematopoietic cells. PMID- 10694516 TI - Circulation Online Only : February 29, 2000. PMID- 10694515 TI - Identification of benzodiazepines in Artemisia dracunculus and Solanum tuberosum rationalizing their endogenous formation in plant tissue. AB - Sterile cultivated plant cell tissues and cell regenerates of several species were tested for their binding affinity to the central human benzodiazepine receptor. Binding activity was found in extracts of Artemisia dracunculus cell tissue (IC(50) = 7 microg/ml) and, to a lesser extent, in plant regenerates of potato herb (Solanum tuberosum). Preparative HPLC led to the isolation of fractions with a significant displacing potency in the benzodiazepine receptor binding assay. Using on-line HPLC-electrospray-tandem mass spectrometry (HPLC-ESI MS/MS) in the "selected reaction monitoring" (SRM) mode, delorazepam and temazepam were found in amounts of about 100 to 200 ng/g cell tissue of Artemisia dracunculus, whereas sterile potato herb contained temazepam and diazepam ranging approximately from 70 to 450 ng/g cell tissue. It is the first report on the endogenous formation of benzodiazepines by plant cells, as any interaction of microorganisms and environmental factors was excluded. PMID- 10694517 TI - Long-term His-bundle pacing and cardiac function. PMID- 10694518 TI - Patent foramen ovale is indicted, but the case hasn't gone to trial. PMID- 10694519 TI - New approaches to pulmonary hypertension: will therapies in mice work in humans? PMID- 10694520 TI - Elevated levels of shed membrane microparticles with procoagulant potential in the peripheral circulating blood of patients with acute coronary syndromes. AB - BACKGROUND: Apoptotic microparticles are responsible for almost all tissue factor activity of the plaque lipid core. We hypothesized that elevated levels of procoagulant microparticles could also circulate in the peripheral blood of patients with recent clinical signs of plaque disruption and thrombosis. METHODS AND RESULTS: We studied 39 patients with coronary heart disease, including 12 patients with stable angina and 27 patients with acute coronary syndromes (ACS), and 12 patients with noncoronary heart disease. We isolated the circulating microparticles by capture with annexin V and determined their procoagulant potential with a prothrombinase assay. The cell origins of microparticles were determined in an additional 22 patients by antigenic capture with specific antibodies. The level of procoagulant microparticles did not differ between stable angina patients and noncoronary patients (10.1+/-1.6 nmol/L phosphatidylserine [PS] equivalent versus 9.9+/-1.6 nmol/L PS equivalent, respectively). However, procoagulant microparticles were significantly elevated in patients with ACS (22.2+/-2.7 nmol/L PS equivalent) compared with other coronary (P<0.01) or noncoronary (P<0.01) patients. Microparticles of endothelial origin were significantly elevated in patients with ACS (P<0.01), which suggests an important role for endothelial injury in inducing the procoagulant potential. CONCLUSIONS: High levels of procoagulant endothelial microparticles are present in the circulating blood of patients with ACS and may contribute to the generation and perpetuation of intracoronary thrombi. PMID- 10694521 TI - Maximally recommended doses of angiotensin-converting enzyme (ACE) inhibitors do not completely prevent ACE-mediated formation of angiotensin II in chronic heart failure. AB - BACKGROUND: The added benefits of angiotensin II type I receptor (AT(1)) blockers (ARBs) to ACE inhibition suggests that recommended doses of ACE inhibitors provide only partial inhibition of ACE in chronic heart failure (CHF). Accordingly, the level of ACE inhibition was assessed by the pressor response to angiotensin (Ang) I in patients who had been treated with recommended doses of ACE inhibitors. METHODS AND RESULTS: Forty-two patients with CHF receiving 40 mg/d of a long-acting ACE inhibitor or 150 mg of captopril were studied. Radial artery systolic pressure (RASP, mm Hg) was monitored noninvasively. The pressor response to ascending doses of Ang I was evaluated in all patients before and after administration of the ARB valsartan. The pressor response to Ang I before and after valsartan was also reevaluated in 11 patients after the dose of ACE inhibitor was doubled for 1 week. RASP increased linearly with significantly ascending doses of Ang I despite treatment with ACE inhibitors. The pressor response to Ang I was blunted significantly by valsartan. Ang I-induced increase in RASP did not correlate with duration of ACE inhibitor therapy. After the dose of ACE inhibitors was doubled, the pressor response to Ang I was no longer different from that noted after valsartan. CONCLUSIONS: Recommended doses of ACE inhibitors do not fully inhibit ACE in CHF. The level of ACE inhibition achieved is not related to duration of ACE inhibitor therapy. Greater ACE inhibition is also achieved at twice the recommended doses of ACE inhibitors. PMID- 10694522 TI - Transcatheter closure of atrial septal defects without fluoroscopy: feasibility of a new method. AB - BACKGROUND: In an effort to reduce x-ray exposure, we developed a technique for transcatheter closure of atrial septal defects under echocardiographic guidance without fluoroscopy. To assess the efficiency of this procedure for routine use, we compared our initial results with those for the conventional procedure. METHODS AND RESULTS: Twenty-two randomly selected patients (median age 18 years; range 2 to 66 years) with atrial septal defects (n=13) or patent foramen ovale (n=9) underwent cardiac catheterization for possible interventional defect closure with echocardiography as the only imaging tool. Median stretched diameter was 9 mm (range 6 to 26 mm); median left-to-right shunt over the atrial septal defects was Qp/Qs=1.8 (range 1.5 to 2.6). An Amplatzer septal occluder was successfully implanted in 19 defects without fluoroscopy and in 3 with the help of radiography. After 1 month, complete defect closure was documented in all patients. Compared with the conventional procedure of a control group of 131 patients, procedure times were not significantly different (88 versus 100 minutes; P=0.09). However, the study group received significantly higher doses of propofol for sedation (9.9 versus 5.6 mg/kg body weight; P=0.002) owing to extended transesophageal echocardiography. CONCLUSIONS: In the majority of patients in whom transcatheter closure of interatrial communications with the Amplatzer septal occluder is possible, the procedure can be safely performed under echocardiographic guidance without fluoroscopy. PMID- 10694523 TI - Identification of patients at increased risk of first unheralded acute myocardial infarction by electron-beam computed tomography. AB - BACKGROUND: There is a clear relationship between absolute calcium scores (CS) and severity of coronary artery disease. However, hard coronary events have been shown to occur across all ranges of CS. METHODS AND RESULTS: We conducted 2 analyses: in group A, 172 patients underwent electron-beam CT (EBCT) imaging within 60 days of suffering an unheralded myocardial infarction. In group B, 632 patients screened by EBCT were followed up for a mean of 32+/-7 months for the development of acute myocardial infarction or cardiac death. The mean patient age and prevalence of coronary calcification were similar in the 2 groups (53+/-8 versus 52+/-9 years and 96% each). In group B, the annualized event rate was 0.11% for subjects with CS of 0, 2.1% for CS 1 to 99, 4.1% for CS 100 to 400, and 4.8% for CS >400, and only 7% of the patients had CS >400. However, mild, moderate, and extensive absolute CSs were distributed similarly between patients with events in both groups (34%, 35%, and 27%, respectively, in group A and 44%, 30%, and 22% in group B). In contrast, the majority of events in both groups occurred in patients with CS >75th percentile (70% in each group). CONCLUSIONS: Coronary calcium is present in most patients who suffer acute coronary events. Although the event rate is greater for patients with high absolute CSs, few patients have this degree of calcification on a screening EBCT. Conversely, the majority of events occur in individuals with high CS percentiles. Hence, CS percentiles constitute a more effective screening method to stratify individuals at risk. PMID- 10694524 TI - Study on the relationship between plasma nitrite and nitrate level and salt sensitivity in human hypertension : modulation of nitric oxide synthesis by salt intake. AB - BACKGROUND: High salt intake suppresses the effect of nitric oxide (NO) in the peripheral resistance vessels in animal models. We tested the hypothesis that the modulation of endogenous NO is related to salt sensitivity in human hypertension. METHODS AND RESULTS: Inpatients with essential hypertension (n=24) were maintained on a normal-salt diet (12 g/d NaCl) for 3 days, a low-salt diet (2 g), a high-salt diet (20 to 23 g), and a low-salt diet for 7 days. Normotensive subjects (n=16) were maintained on the first 2 salt diets. The hypertensive patients whose average 24-hour blood pressure was increased by >5% by salt loading were assigned to group 1 (n=8) and the others to group 2 (n=16). Nitrate plus nitrite (NO(x)) was measured by the Griess method, and asymmetrical dimethylarginine (ADMA) by high-performance liquid chromatography. The plasma NO(x) level during the normal-salt diet was lower in group 1 than in group 2 and the normotensive group. After salt loading, the plasma NO(x) level was decreased and reversed after the second salt restriction. Plasma ADMA level was increased after salt loading and decreased after salt restriction. The change in plasma NO(x) level was correlated inversely with those in blood pressure (r=-0.59, P=0.0007) and plasma ADMA level (r=-0.64, P=0.003) after salt loading and restriction. CONCLUSIONS: Modulation of NO synthesis by salt intake may be involved in a mechanism for salt sensitivity in human hypertension, presumably via the change in ADMA. PMID- 10694525 TI - Influence of the menstrual cycle on sympathetic activity, baroreflex sensitivity, and vascular transduction in young women. AB - BACKGROUND: Our goal was to test sympathetic and cardiovagal baroreflex sensitivity and the transduction of sympathetic traffic into vascular resistance during the early follicular (EF) and midluteal (ML) phases of the menstrual cycle. METHODS AND RESULTS: Sympathetic baroreflex sensitivity was assessed by lowering and raising blood pressure with intravenous bolus doses of sodium nitroprusside and phenylephrine. It was defined as the slope relating muscle sympathetic nerve activity (MSNA; determined by microneurography) and diastolic blood pressure. Cardiovagal baroreflex sensitivity was defined as the slope relating R-R interval and systolic blood pressure. Vascular transduction was evaluated during ischemic handgrip exercise and postexercise ischemia, and it was defined as the slope relating MSNA and calf vascular resistance (determined by plethysmography). Resting MSNA (EF, 1170+/-151 U/min; ML, 2252+/-251 U/min; P<0.001) and plasma norepinephrine levels (EF, 240+/-21 pg/mL; ML, 294+/-25 pg/mL; P=0. 025) were significantly higher in the ML than in the EF phase. Furthermore, sympathetic baroreflex sensitivity was greater during the ML than the EF phase in every subject (MSNA/diastolic blood pressure slopes: EF, -4.15; FL, -5.42; P=0.005). No significant differences in cardiovagal baroreflex sensitivity or vascular transduction were observed. CONCLUSIONS: The present study suggests that the hormonal fluctuations that occur during the normal menstrual cycle may alter sympathetic outflow but not the transduction of sympathetic activity into vascular resistance. PMID- 10694526 TI - Permanent, direct His-bundle pacing: a novel approach to cardiac pacing in patients with normal His-Purkinje activation. AB - BACKGROUND: Direct His-bundle pacing (DHBP) produces synchronous ventricular depolarization and improved cardiac function relative to apical pacing. Although it has been performed transiently in the electrophysiology laboratory and persistently in open-chested canines, permanent DHBP in humans has not been achieved. METHODS AND RESULTS: A total of 18 patients aged 69+/-10 years who had a history of chronic atrial fibrillation, dilated cardiomyopathy, and normal activation (ie, QRS< or =120 ms) were screened for permanent DHBP using an electrophysiology catheter. In 14 patients, the His bundle could be reliably stimulated. Of these 14, permanent DHBP using a fixed screw-in lead was successful in 12 patients. Radiofrequency atrioventricular node ablation was performed in patients exhibiting a fast ventricular response. All patients received single-chamber rate-responsive pacemakers. Acute pacing thresholds were 2.4+/-1.0 V at a pulse duration of 0.5 ms. Lead complications included exit block requiring reoperative adjustment and gross lead dislodgment. Echocardiographic improvement in heart function was shown by reductions in the left ventricular end diastolic dimension from 59+/-8 to 52+/-6 mm (P1 hour, for a total of 2697 hours of continuous detection of AT/AF. During Holter monitoring, the sensitivity of the detection of AT/AF (116 hours) was 100%; the specificity of the detection of non AT/AF rhythms (1290 hours) was 99.99%. Of 166 appropriate episodes detected as AT, 45% were terminated by antitachycardia pacing. CONCLUSIONS: A new ICD detects AT/AF accurately and continuously. Therapy may be programmed for long-duration AT/AF, with a low risk of underdetection. Discrimination of AT from AF permits successful pacing therapy for a significant fraction of AT. PMID- 10694528 TI - Oscillatory patterns in sympathetic neural discharge and cardiovascular variables during orthostatic stimulus. AB - BACKGROUND: We tested the hypothesis that a common oscillatory pattern might characterize the rhythmic discharge of muscle sympathetic nerve activity (MSNA) and the spontaneous variability of heart rate and systolic arterial pressure (SAP) during a physiological increase of sympathetic activity induced by the head up tilt maneuver. METHODS AND RESULTS: Ten healthy subjects underwent continuous recordings of ECG, intra-arterial pressure, respiratory activity, central venous pressure, and MSNA, both in the recumbent position and during 75 degrees head-up tilt. Venous samplings for catecholamine assessment were obtained at rest and during the fifth minute of tilt. Spectrum and cross-spectrum analyses of R-R interval, SAP, and MSNA variabilities and of respiratory activity provided the low (LF, 0.1 Hz) and high frequency (HF, 0.27 Hz) rhythmic components of each signal and assessed their linear relationships. Compared with the recumbent position, tilt reduced central venous pressure, but blood pressure was unchanged. Heart rate, MSNA, and plasma epinephrine and norepinephrine levels increased, suggesting a marked enhancement of overall sympathetic activity. During tilt, LF(MSNA) increased compared with the level in the supine position; this mirrored similar changes observed in the LF components of R-R interval and SAP variabilities. The increase of LF(MSNA) was proportional to the amount of the sympathetic discharge. The coupling between LF components of MSNA and R-R interval and SAP variabilities was enhanced during tilt compared with rest. CONCLUSIONS: During the sympathetic activation induced by tilt, a similar oscillatory pattern based on an increased LF rhythmicity characterized the spontaneous variability of neural sympathetic discharge, R-R interval, and arterial pressure. PMID- 10694529 TI - Percutaneous closure of patent foramen ovale in patients with paradoxical embolism: long-term risk of recurrent thromboembolic events. AB - BACKGROUND: Patients with a patent foramen ovale (PFO) and paradoxical embolism are at risk for recurrent thromboembolic events. This study investigated the long term risk of recurrent thromboembolic events in patients with PFO and paradoxical embolism after percutaneous PFO closure. METHODS AND RESULTS: Since 1994, a total of 80 patients with PFO and at least 1 paradoxical embolic event (transient ischemic attack [TIA], cerebrovascular accident [CVA], peripheral embolism) have undergone percutaneous PFO closure with 5 different devices. There were 30 women and 50 men, with a mean age of 52+/-12 years. Sixty patients had only a PFO, whereas 20 patients had both a PFO and an atrial septal aneurysm. The implantation procedure was successful in 78 patients (98%). During 5 years of follow-up (mean, 1.6+/-1.4 years; range, 0.1 to 5.0 years), the actuarial annual risk to suffer a recurrent thromboembolic event was 2.5% for TIA, 0% for CVA, 0.9% for peripheral emboli, and 3.4% for the combined end point of TIA, CVA, or peripheral embolism. A postprocedural shunt was a predictor of recurrent paradoxical embolism (RR, 4.2; 95% CI, 1.1 to 17.8; P=0.03). The risk for recurrent thromboembolic events in patients with both atrial septal aneurysm and PFO was not significantly increased compared with patients with only PFO (RR, 1.0; 95% CI, 0.2 to 4.7; P=0.95). CONCLUSIONS: Percutaneous PFO closure appears to be a promising technique in the prevention of recurrent systemic thromboembolism in patients with a PFO after a first event. Prospective studies comparing percutaneous PFO closure with antithrombotic medications or surgery must define its therapeutic value. PMID- 10694530 TI - Alteration in left ventricular diastolic filling and accumulation of myocardial collagen at insulin-resistant prediabetic stage of a type II diabetic rat model. AB - BACKGROUND: Considerable controversy exists regarding impairment of cardiac function in diabetes mellitus (DM). We investigated the serial changes in left ventricular (LV) histopathology and LV filling dynamics in Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which have been established as an animal model of type II DM. METHODS AND RESULTS: In 54 OLETF and 54 non-DM rats, body weight, blood pressure, heart rate, and transmitral pulsed Doppler examinations were performed from 5 to 47 weeks of age. An oral glucose tolerance test was performed at 10, 20, and 30 weeks of age. The hearts were excised for histopathology, including immunohistochemistry and histomorphometry of collagen, and measurement of hydroxyproline at baseline and each stage of developing DM. In the prediabetic stage (15 weeks of age), in which fast blood glucose remained normal, OLETF rats manifested mild obesity, postprandial hyperglycemia, and hyperinsulinemia, and early diastolic transmitral inflow exhibited prolonged deceleration time (OLETF, 59+/-10 ms versus non-DM, 49+/-8 ms, P<0.01) and low peak velocity (OLETF, 73+/ 11 cm/s versus non-DM, 88+/-11 cm/s, P<0.01). Histopathology revealed extracellular fibrosis and abundant transforming growth factor-beta(1) receptor II in LV myocytes of OLETF rats. At 15 weeks of age, the ratio of collagen area/visual field of LV wall in OLETF rats (8.3+/-1.3%) was larger than that in non-DM rats (4.9+/-1.8%, P<0.0001), and the collagen content/dry tissue weight ratio of heart was significantly higher in OLETF (2. 0+/-0.5 mg/g) than non-DM (1.3+/-0.2 mg/g, P<0.01) rats. CONCLUSIONS: A metabolic abnormality present in the prestage of type II DM may produce LV fibrosis and alteration in cardiac function. PMID- 10694531 TI - Interleukin-10 inhibits intimal hyperplasia after angioplasty or stent implantation in hypercholesterolemic rabbits. AB - BACKGROUND: Intimal hyperplasia after stent implantation is the main cause of in stent restenosis. Activated monocytes play a key role in intimal growth. The anti inflammatory cytokine interleukin-10 (IL-10) is a potent monocyte deactivator, endogenously produced in the atherosclerotic plaque. We tested the hypothesis that exogenous IL-10 may limit postangioplasty intimal hyperplasia after balloon angioplasty or stenting. METHODS AND RESULTS: Hypercholesterolemic rabbits were treated with recombinant human IL-10 (rhuIL-10) for 3 days after balloon angioplasty or 28 days after stent implantation. High IL-10 serum levels and intense deactivation of circulating monocytic cells, assessed by inhibition of IL 1beta release by lipopolysaccharide-stimulated whole blood, were detected for at least 8 hours after rhuIL-10 intravenous injection (ELISA). Morphometric analyses, performed 28 days after injury, indicated that rhuIL-10 reduced intimal growth by approximately 50% after balloon angioplasty or stenting, resulting in more preserved lumen in stented arteries. Moreover, rhuIL-10 reduced macrophage infiltration by 67% and proliferative activity by 81% in the intima and the media. No toxic effect was detected except minor changes in blood cell count. CONCLUSIONS: The anti-inflammatory cytokine rhuIL-10 reduces postinjury intimal hyperplasia. The potent attenuation of in-stent intimal growth by rhuIL-10 and its favorable toxicity profile suggest that rhuIL-10 may be useful in the prevention of in-stent restenosis. PMID- 10694532 TI - Persistent changes in myocardial glucose metabolism in vivo during reperfusion of a limited-duration coronary occlusion. AB - BACKGROUND: Rapid reperfusion of an occluded coronary artery salvages regional mechanical function, but this benefit may not be realized for hours or days because of postischemic stunning. Recovery from stunning is incompletely understood but may involve adaptive changes in heart glucose metabolism. METHODS AND RESULTS: To examine whether reversible coronary occlusion produces sustained changes in regional glucose metabolism in vivo, we performed a 20-minute left coronary artery occlusion followed by 24 hours of open-artery reperfusion in intact rats. Coronary occlusion produced stunning of the anterolateral left ventricle that resolved over 24 hours. When examined at 24 hours, reperfused regions were fully contractile and viable by vital staining and microscopy but demonstrated 25% reduction in blood flow and 50% increased uptake of circulating glucose, as estimated by in vivo [(13)N]NH(3) and [(18)F]fluorodeoxyglucose (FDG) tracer uptake. Reperfused regions had largely inactive glycogen synthase, low rates of glycogen synthesis, and persistent 50% glycogen depletion but increased flux of plasma [1-(13)C]glucose into myocardial [3-(13)C]alanine, indicating preferential shunting of imported glucose away from storage and into glycolysis. CONCLUSIONS: Sustained increases in regional glycolytic consumption of circulating glucose occur during reperfusion of a limited-duration coronary occlusion. This suggests a role for glycolytic ATP in the recovery from postischemic stunning in vivo. Furthermore, [(13)N]NH(3) /FDG regional mismatch may constitute a clinically accessible late metabolic signature of regional myocardial ischemia. PMID- 10694534 TI - Resistance to endotoxin shock in transgenic mice overexpressing endothelial nitric oxide synthase. AB - BACKGROUND: Nitric oxide (NO) plays a central role in the pathogenesis of septic shock. However, the role of the NO produced by endothelial NO synthase (eNOS) in septic shock is still unclear. We examined the effect of chronic eNOS overexpression and the role of eNOS-derived NO in lipopolysaccharide (LPS) induced septic shock using eNOS transgenic (Tg) mice. METHODS AND RESULTS: LPS was intraperitoneally injected into Tg and control mice. No differences existed in the peak plasma nitrate and nitrate levels induced by LPS between the 2 genotypes. In LPS-treated control mice, blood pressure progressively declined and reached 60% of basal levels (from 97+/-3 to 59+/-3 mm Hg) 24 hours after LPS injection. In contrast, the blood pressure of LPS-treated Tg mice fell only 15% from basal levels (from 84+/-4 to 71+/-4 mm Hg) after the first 6 hours and, thereafter, it remained at this level. LPS-induced increases in the expression of the mRNA of both vascular cell adhesion molecule-1 and intracellular adhesion molecule-1 in the lungs were significantly lower in Tg mice than in control mice. LPS-induced pulmonary leukocyte infiltration and increases in lung water content were also significantly attenuated in Tg mice. Histological examination revealed that lung injury after LPS injection was milder in Tg mice. Furthermore, Tg mice exhibited enhanced survival from LPS-induced septic shock compared with control mice. CONCLUSIONS: Chronic eNOS overexpression in the endothelium of mice resulted in resistance to LPS-induced hypotension, lung injury, and death. These effects are associated with the reduced vascular reactivity to NO and the reduced anti-inflammatory effects of NO. PMID- 10694533 TI - In vivo gene transfer of prepro-calcitonin gene-related peptide to the lung attenuates chronic hypoxia-induced pulmonary hypertension in the mouse. AB - BACKGROUND: Calcitonin gene-related peptide (CGRP) is believed to play an important role in maintaining low pulmonary vascular resistance (PVR) and in modulating pulmonary vascular responses to chronic hypoxia; however, the effects of adenovirally mediated gene transfer of CGRP on the response to hypoxia are unknown. METHODS AND RESULTS: In the present study, an adenoviral vector encoding prepro-CGRP (AdRSVCGRP) was used to examine the effects of in vivo gene transfer of CGRP on increases in PVR, right ventricular mass (RVM), and pulmonary vascular remodeling that occur in chronic hypoxia in the mouse. Intratracheal administration of AdRSVCGRP, followed by 16 days of chronic hypoxia (FIO(2) 0.10), increased lung CGRP and cAMP levels. The increase in pulmonary arterial pressure (PAP), PVR, RVM, and pulmonary vascular remodeling in response to chronic hypoxia was attenuated in animals overexpressing prepro-CGRP, whereas systemic pressure was not altered while in chronically hypoxic mice, angiotensin II and endothelin-1-induced increases in PAP were reduced, whereas decreases in PAP in response to CGRP and adrenomedullin were not changed and decreases in PAP in response to a cAMP phosphodiesterase inhibitor were enhanced by AdRSVCGRP. CONCLUSIONS: In vivo CGRP lung gene transfer attenuates the increase in PVR and RVM, pulmonary vascular remodeling, and pressor responses in chronically hypoxic mice, suggesting that CGRP gene transfer alone and with a cAMP phosphodiesterase inhibitor may be useful for the treatment of pulmonary hypertensive disorders. PMID- 10694535 TI - Images in cardiovascular medicine. Abciximab-associated pseudothrombocytopenia. PMID- 10694536 TI - Diagnosis of an intracoronary thrombus with intravascular ultrasound. PMID- 10694537 TI - Cardiac interleukin-6 in ischemic myocardium. PMID- 10694538 TI - Alpha-adrenergic blockade in myocardial infarction. PMID- 10694539 TI - The nature of the statins. PMID- 10694540 TI - Sildenafil/nitrate interaction. PMID- 10694541 TI - Long-term results of percutaneous balloon mitral valvuloplasty using the Inoue balloon catheter technique. PMID- 10694542 TI - Myelodysplasia and acute leukemia after autologous bone marrow transplantation. PMID- 10694543 TI - Therapy-related myelodysplasia and secondary acute myelogenous leukemia after high-dose therapy with autologous hematopoietic progenitor-cell support for lymphoid malignancies. AB - PURPOSE: To evaluate the incidence of and risk factors for therapy-related myelodysplasia (tMDS) and secondary acute myelogenous leukemia (sAML), after high dose therapy (HDT) with autologous bone marrow or peripheral-blood progenitor cell support, in patients with non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Between January 1985 and November 1996, 230 patients underwent HDT comprising cyclophosphamide therapy and total-body irradiation, with autologous hematopoietic progenitor-cell support, as consolidation of remission. With a median follow-up of 6 years, 27 (12%) developed tMDS or sAML. RESULTS: Median time to development of tMDS or sAML was 4.4 years (range, 11 months to 8.8 years) after HDT. Karyotyping (performed in 24 cases) at diagnosis of tMDS or sAML revealed complex karyotypes in 18 patients. Seventeen patients had monosomy 5/5q , 15 had -7/7q-, seven had -18/18q-, seven had -13/13q-, and four had -20/20q-. Twenty-one patients died from complications of tMDS or sAML or treatment for tMDS or sAML, at a median of 10 months (range, 0 to 26 months). Sixteen died without evidence of recurrent lymphoma. Six patients were alive at a median follow-up of 6 months (range, 2 to 22 months) after diagnosis of tMDS or sAML. On multivariate analysis, prior fludarabine therapy (P =.009) and older age (P =.02) were associated with the development of tMDS or sAML. Increased interval from diagnosis to HDT and bone marrow involvement at diagnosis were of borderline significance (P =.05 and.07, respectively). CONCLUSION: tMDS and sAML are serious complications of HDT for NHL and are associated with very poor prognosis. Alternative strategies for reducing their incidence and for treatment are needed. PMID- 10694544 TI - Low-dose 5-aza-2'-deoxycytidine, a DNA hypomethylating agent, for the treatment of high-risk myelodysplastic syndrome: a multicenter phase II study in elderly patients. AB - PURPOSE: 5-Aza-2'-deoxycytidine (decitabine; DAC) is a DNA hypomethylating agent that has shown a 50% response rate in a small phase II study in elderly patients with high-risk myelodysplastic syndrome. We performed a second, multicenter phase II study in a larger group of patients to confirm our findings and to study the toxicity of DAC. PATIENTS AND METHODS: Between June 1996 and September 1997, 66 patients (median age, 68 years) from seven centers received DAC 45 mg/m(2)/d for 3 days every 6 weeks. For patients in whom a complete response (CR) was reached after two courses, two further cycles were administered as consolidation therapy. In case of a stable disease situation, improvement, or a partial response (PR), a maximum of six cycles was administered. The primary end points were response rate and toxicity. The secondary end points were response duration, survival from the start of therapy, and overall survival. RESULTS: The observed overall response rate was 49%, with a 64% response rate in the patients with an International Prognostic Scoring System (IPSS) high-risk score. The actuarial median response duration was 31 weeks, with a response duration of 39 weeks and 36 weeks for patients who reached a PR or CR, respectively. The actuarial median survival time from the time of diagnosis was 22 months and from the start of therapy was 15 months. For the IPSS high-risk group, the median survival time was 14 months. The median progression-free survival time was 25 weeks. Myelosuppression was rather common, and the treatment-related mortality rate was 7% and was primarily associated with pancytopenia and infection. Significant responses were observed with regard to megakaryopoiesis, with increases in platelet counts having already occurred after one cycle of DAC therapy in the majority of the responding patients. CONCLUSION: We were able to confirm our previous observation that DAC therapy was effective in half of the studied patients with high-risk myelodysplastic syndrome and is especially active in the patients with the worst prognoses. Myelosuppression was the only major adverse effect observed. PMID- 10694545 TI - Allogeneic bone marrow transplantation for therapy-related myelodysplastic syndrome and acute myeloid leukemia: a long-term study of 70 patients-report of the French society of bone marrow transplantation. AB - PURPOSE: To identify predictive factors of survival, relapse, and transplantation related mortality (TRM) among patients with therapy-related myelodysplastic syndrome (t-MDS) or acute leukemia (t-AML) who underwent allogeneic bone marrow transplantation (BMT). PATIENTS AND METHODS: From 1980 to 1998, 70 patients underwent allogeneic BMT for t-MDS (n = 31) or t-AML (n = 39) after prior cytotoxic exposure. Thirty-three patients had received induction-type chemotherapy before BMT. At the time of transplantation, there were 24 patients in complete remission (CR) and 46 with active disease. RESULTS: With a median follow-up of 7.9 years (range, 1.1 to 18.8 years) after BMT, 16 patients are alive, whereas 19 died of relapse, 34 of TRM, and one of relapse of the primary disease. The estimated 2-year overall survival, event-free survival, relapse, and TRM rates were 30% (95% confidence interval [CI], 19% to 40%), 28% (95% CI, 18% to 39%), 42% (95% CI, 26% to 57%), and 49% (95% CI, 36% to 62%), respectively. In multivariable analysis, age greater than 37 years, male sex, positive recipient cytomegalovirus (CMV) serology, absence of CR at BMT, and intensive schedules used for conditioning were associated with poor outcome. CONCLUSION: BMT is an effective treatment for patients with t-MDS or t-AML who have responsive disease and, in particular, who have no poor-risk cytogenetic features. The poor results of the other patients, especially those with active disease at BMT, emphasize the need to delineate indications and perform prospective protocols. PMID- 10694546 TI - Assessment of the stanford V regimen and consolidative radiotherapy for bulky and advanced Hodgkin's disease: Eastern Cooperative Oncology Group pilot study E1492. AB - PURPOSE: This study was performed, in a multi-institutional setting, to evaluate the efficacy and feasibility of the Stanford V chemotherapy regimen plus radiotherapy to bulky Hodgkin's disease sites. PATIENTS AND METHODS: A two-stage design was implemented in a phase II study involving 47 patients with bulky mediastinal stage I/II or stage III/IV Hodgkin's disease. Twelve weeks of the Stanford V chemotherapy regimen were given with consolidative radiotherapy (36 Gy) to lymph nodes >/= 5 cm and/or macroscopic splenic disease. Treatment was administered in one of five institutions participating in the Eastern Cooperative Oncology Group. RESULTS: With a median follow-up of 4.8 years, 45 patients are alive and 40 have been continuously disease-free. The estimated freedom from progression was 87% at 2 years and 85% at 5 years. Overall survival was 96% at 2 and 5 years. There was one death from Hodgkin's disease and one death from an M5 acute leukemia. Six of seven relapsed patients received high-dose therapy and autologous stem-cell transplantation. The freedom from second progression for the seven relapsed patients was estimated at 98% at 3 years. CONCLUSION: Stanford V chemotherapy and consolidative radiotherapy to bulky disease is effective in bulky and advanced Hodgkin's disease in a multi-institutional setting. On this basis, an Intergroup study comparing doxorubicin, bleomycin, vinblastine, and dacarbazine with the Stanford V regimen has been initiated. PMID- 10694547 TI - Prospective randomized comparison of single-dose versus hyperfractionated total body irradiation in patients with hematologic malignancies. AB - PURPOSE: Fractionated total-body irradiation (HTBI) is considered to induce less toxicity to normal tissues and probably has the same efficacy as single-dose total-body irradiation (STBI) in patients with acute myeloid leukemia. We decided to determine whether this concept can be applied to a large number of patients with various hematologic malignancies using two dissimilar fractionation schedules. PATIENTS AND METHODS: Between December 1986 and October 1994, 160 patients with various hematologic malignancies were randomized to receive either a 10-Gy dose of STBI or 14.85-Gy dose of HTBI. RESULTS: One hundred forty-seven patients were assessable. The 8-year overall survival rate and cause-specific survival rate in the STBI group was 38% and 63.5%, respectively. Overall survival rate and cause-specific survival rate in the HTBI group was 45% and 77%, respectively. The incidence of interstitial pneumonitis was similar in both groups. However, the incidence of veno-occlusive disease (VOD) of the liver was significantly higher in the STBI group. In the multivariate analysis with overall survival as the end point, the female sex was an independent favorable prognostic factor. On the other hand, when cause-specific survival was considered as the end point, the multivariate analysis demonstrated that sex and TBI were independent prognostic factors. CONCLUSION: The efficacy of HTBI is probably higher than that of STBI. Both regimens induce similar toxicity with the exception of VOD of the liver, the incidence of which is significantly more pronounced in the STBI group. PMID- 10694548 TI - Phase I study of fludarabine plus cyclophosphamide in patients with previously untreated low-grade lymphoma: results and and long-term follow-up--a report from the Eastern Cooperative Oncology Group. AB - PURPOSE: To determine the toxicity and recommended phase II doses of the combination of fludarabine plus cyclophosphamide in chemotherapy-naive patients with low-grade lymphoma. PATIENTS AND METHODS: Previously untreated patients with low-grade lymphoma were entered onto dosing cohorts of four patients each. The cyclophosphamide dose, given on day 1, was increased from 600 to 1, 000 mg/m(2). Fludarabine 20 mg/m(2) was administered on days 1 through 5. The first eight patients were treated every 21 days; later patients were treated every 28 days. Prophylactic antibiotics were required. RESULTS: Prolonged cytopenia and pulmonary toxicity each occurred in three of eight patients treated every 3 weeks. The 19 patients treated every 28 days, who were given granulocyte colony stimulating factor as indicated, did not have undue nonhematologic toxicity. Dose limiting toxicity was hematologic. At the recommended phase II/III dose (cyclophosphamide 1,000 mg/m(2)), grade 4 neutropenia was observed in 17% of all cycles and 31% of first cycles. Grade 3 or 4 thrombocytopenia was seen in only 1% of all cycles. The median number of cycles per patient was six (range, two to 11) for all patients enrolled. The response rate was 100% of 27 patients entered; 89% achieved a complete and 11% a partial response. Nineteen of 22 patients with bone marrow involvement had clearing of the marrow. Median duration of follow-up was more than 5 years; median overall and disease-free survival times have not been reached. Kaplan-Meier estimated 5-year overall survival and disease-free survival rates were 66% and 53%, respectively. CONCLUSION: The recommended dosing for this combination in patients with previously untreated low-grade lymphoma is cyclophosphamide 1, 000 mg/m(2) day 1 and fludarabine 20 mg/m(2) days 1 through 5. The regimen has a high level of activity, with prolonged complete remissions providing 5-year overall and disease-free survival rates as high as those reported for other therapeutic approaches in untreated patients. PMID- 10694549 TI - Pharmacokinetics of nelarabine and 9-beta-D-arabinofuranosyl guanine in pediatric and adult patients during a phase I study of nelarabine for the treatment of refractory hematologic malignancies. AB - PURPOSE: To characterize the pharmacokinetics of nelarabine (506U78), the water soluble prodrug of 9-beta-D-arabinofuranosyl guanine (ara-G), and ara-G in pediatric and adult patients with refractory hematologic malignancies. Ara-G is phosphorylated within leukemic cells to form ara-G triphosphate (ara-GTP), which acts to terminate DNA chain elongation, resulting in cell death. PATIENTS AND METHODS: The pharmacokinetics of nelarabine and/or ara-G were evaluated in 71 patients (25 pediatric and 46 adult patients) on the first day of therapy. Blood was collected at specified times for the determination of plasma nelarabine and ara-G concentrations. RESULTS: There were no statistically significant differences in the pharmacokinetics of nelarabine between any of the groups of patients. The harmonic mean half-life (t1/2) of nelarabine in pediatric and adult patients was 14.1 minutes and 16.5 minutes, respectively. The maximum concentrations (C(max)) of ara-G occurred at or near the end of the nelarabine infusion. The C(max) of ara-G ranged from 11.6 micromol/L to 308.7 micromol/L at nelarabine doses of 5 to 75 mg/kg and was linearly related to the nelarabine dose. No statistically significant differences were noted for the pharmacokinetic parameter estimates of ara-G between adult male and female patients. In children versus adults, the dose-normalized C(max), time of the C(max), and the steady state volume of distribution of ara-G were similar. However, the clearance of ara G was higher in pediatric patients (0.312 L.h(-1).kg(-1)) as compared with adult patients (0. 213 L.h(-1).kg(-1)) (P <.001). The t1/2 of ara-G was shorter in pediatric patients as compared with adult patients (2.1 hours v 3.0 hours; P <.01). CONCLUSION: Nelarabine is an effective prodrug of ara-G, allowing systemic concentrations of ara-G that result in clinical activity. PMID- 10694550 TI - Cognitive and academic functioning in survivors of pediatric bone marrow transplantation. AB - PURPOSE: To evaluate cognitive and academic functioning in survivors of pediatric bone marrow transplants (BMTs) at 1 and 3 years after a BMT. PATIENTS AND METHODS: In a prospective, longitudinal design, patients underwent a comprehensive battery of neurocognitive measures before admission for transplantation and at 1, 3, and 5 years after a BMT. This article describes a cohort of 102 survivors with follow-up data available for 1 year after a BMT, including 54 survivors with follow-up available for 3 years. This represents the largest cohort of pediatric BMT survivors yet reported in a prospective study. RESULTS: In the cohort as a whole, there were no significant changes on global measures of intelligence (intelligence quotient [IQ]) and academic achievement at either 1 or 3 years after a BMT, despite adequate power to detect an IQ change of three points or greater. Likewise, performance on specific tests of neuropsychologic function remained stable. No significant differences were observed between patients whose conditioning regimen included total-body irradiation (TBI) and those whose did not. The primary predictor of neurocognitive outcome was patient age, with younger patients more likely to show declines over time. The subset of patients who were less than 3 years of age at the time of transplantation seemed to be particularly vulnerable to cognitive sequelae. CONCLUSION: The use of BMTs with or without TBI entails minimal risk of late neurocognitive sequelae in patients who are 6 years of age or older at the time of transplantation. However, patients who are less than 6 years of age at the time of transplantation, and particularly those less than 3 years of age, seem to be at some risk of cognitive declines. PMID- 10694551 TI - "Low-risk" prediction rule for pediatric oncology patients presenting with fever and neutropenia. AB - PURPOSE: To prospectively derive and validate a clinical prediction rule to allow a more tailored approach to the management of pediatric oncology outpatients presenting with fever and neutropenia. PATIENTS AND METHODS: The clinical prediction rule was derived over a 1-year period and then validated over the following 8 months in a new set of fever and neutropenia episodes. Patients were excluded if they presented with comorbidity or an abnormal chest x-ray (CXR). RESULTS: Significant bacterial infection (SBI; defined as any blood or urine culture positive for bacteria, interstitial or lobar consolidation on CXR, or unexpected death from infection) was documented in 43 of the 227 episodes. Multivariate analysis found four significant factors: bone marrow disease, general appearance unwell on initial examination, monocyte count less than 0.1 x 10(9)/L, and peak oral or oral equivalent temperature greater than 39 degrees C. Only the monocyte count contributed to determining a low-risk group, excluding SBI with 84% sensitivity (95% confidence interval [CI], 61% to 100%), 42% specificity (95% CI, 38% to 46%), and a negative predictive value of 92% (95% CI, 76% to 100%). If the monocyte count was >/= 0.1 x 10(9)/L at the time of presentation (low risk), the incidences of SBI and bacteremia were 8% and 5%, respectively, versus 25% and 17% in the high-risk group. When validated in a new population of 136 episodes of fever and neutropenia, the incidences of SBI and bacteremia in the low-risk group were 12% and 5%, respectively, and 25% and 19% in the high-risk group. CONCLUSION: Pediatric oncology outpatients with fever and neutropenia who present with an initial monocyte count of >/= 0.1 x 10(9)/L and do not have comorbidity or an abnormal CXR at the time of presentation are at lower risk for SBI and can be considered for less aggressive initial therapy. PMID- 10694552 TI - Proton magnetic resonance spectroscopic imaging in children with recurrent primary brain tumors. AB - PURPOSE: Proton magnetic resonance spectroscopic imaging ((1)H-MRSI) is a noninvasive technique for spatial characterization of biochemical markers in tissues. We measured the relative tumor concentrations of these biochemical markers in children with recurrent brain tumors and evaluated their potential prognostic significance. PATIENTS AND METHODS: (1)H-MRSI was performed on 27 children with recurrent primary brain tumors referred to our institution for investigational drug trials. Diagnoses included high-grade glioma (n = 10), brainstem glioma (n = 7), medulloblastoma/peripheral neuroectodermal tumor (n = 6), ependymoma (n = 3), and pineal germinoma (n = 1). (1)H-MRSI was performed on 1. 5-T magnetic resonance imagers before treatment. The concentrations of choline (Cho) and N-acetyl-aspartate (NAA) in the tumor and normal brain were quantified using a multislice multivoxel method, and the maximum Cho:NAA ratio was determined for each patient's tumor. RESULTS: The maximum Cho:NAA ratio ranged from 1.1 to 13.2 (median, 4.5); the Cho:NAA ratio in areas of normal-appearing brain tissue was less than 1.0. The maximum Cho:NAA ratio for each histologic subtype varied considerably; approximately equal numbers of patients within each tumor type had maximum Cho:NAA ratios above and below the median. Patients with a maximum Cho:NAA ratio greater than 4.5 had a median survival of 22 weeks, and all 13 patients died by 63 weeks. Patients with a Cho:NAA ratio less than or equal to 4.5 had a projected survival of more than 50% at 63 weeks. The difference was statistically significant (P =.0067, log-rank test). CONCLUSION: The maximum tumor Cho:NAA ratio seems to be predictive of outcome in children with recurrent primary brain tumors and should be evaluated as a prognostic indicator in newly diagnosed childhood brain tumors. PMID- 10694553 TI - TrkC expression predicts good clinical outcome in primitive neuroectodermal brain tumors. AB - PURPOSE: To identify biologic prognostic factors in childhood primitive neuroectodermal tumors (PNET), including medulloblastoma, that accurately define patient groups with sufficiently good prognosis to permit a reduction in treatment intensity. PATIENTS AND METHODS: We determined expression levels of the neurotrophin receptor TrkC mRNA in formalin-fixed tumor samples from 87 well characterized PNET patients using in situ hybridization. Comparison of TrkC mRNA expression levels with clinical and other laboratory variables was performed using univariate and multivariate Cox regression analysis. RESULTS: High TrkC mRNA expression was found to be associated more with higher 5-year cumulative survival rate than was low TrkC mRNA expression (89% v 46%, respectively). When compared with established clinical prognostic factors and laboratory variables of potential prognostic significance, TrkC mRNA expression, by univariate analysis, was found to be the single most powerful predictor of outcome (hazards ratio, 4.81; P <.00005), exceeding all clinical prognostic factors. In multivariate analysis, the hazards ratio remained significant (P <.00005). CONCLUSION: High TrkC mRNA expression in PNET is a powerful independent predictor of favorable clinical outcome. Assessment of TrkC mRNA levels may aid in treatment planning for patients with PNETs and should be incorporated prospectively into PNET clinical trials. PMID- 10694554 TI - Serum human glandular kallikrein-2 protease levels predict the presence of prostate cancer among men with elevated prostate-specific antigen. AB - PURPOSE: We hypothesize that serum human glandular kallikrein-2 (hK2) levels predict the presence of prostate cancer among men prescreened by prostate specific antigen (PSA). PATIENTS AND METHODS: We conducted a cross-sectional study of 324 men who had no history of prostate cancer and who were referred for prostate biopsy. PSA and hK2 levels were measured using specific nonisotopic immunometric techniques. Cases were patients who were diagnosed with adenocarcinoma of the prostate from biopsy, and controls were patients who had no evidence of cancer from biopsy. The odds ratio for detection of prostate cancer was determined for hK2 measurements, controlling for age, total-PSA level, digital rectal examination, and symptoms of urinary obstruction. RESULTS: Of 324 men, 159 (49.1%) had cancer. Mean hK2 levels and hK2:free-PSA ratios were significantly higher in cases than in controls (1.18 v 0.53 ng/mL, respectively, for hK2, P =.0001; 1.17 v 0.62 for hK2:free-PSA ratio, P =.0001). The crude odds ratio for prostate cancer detection for patients in the highest quartile of hK2 level was 5.83 (95% confidence interval [CI], 2.8 to 12.1; P =.0001) compared with patients in the lowest quartile. The adjusted odds ratio was 6.72 (95% CI, 2.9 to 15.6; P =.0001). Similarly, the crude and adjusted odds ratios for prostate cancer detection using the hK2:free-PSA ratio were 7.36 (95% CI, 3.6 to 15.1; P =.0001) and 8.06 (95% CI, 3. 7 to 17.4; P =.0001), respectively. These odds ratios were higher than that observed for prostate cancer detection by total PSA level (2.73; P =.03). CONCLUSION: Among men prescreened with PSA for prostate cancer, patients with high hK2 measurements have a five- to eight-fold increase in risk for prostate cancer, adjusting for PSA level and other established risk factors. hK2 measurements may be a useful adjunct to PSA in improving patient selection for prostate biopsy. PMID- 10694555 TI - Androgen deprivation and four courses of fixed-schedule suramin treatment in patients with newly diagnosed metastatic prostate cancer: A Southwest Oncology Group Study. AB - PURPOSE: To assess the feasibility of administering a combination of suramin and hydrocortisone in addition to androgen deprivation in a cooperative group setting; to assess the feasibility of treatment with multiple courses of suramin; and to assess progression-free and overall survival in patients with newly diagnosed metastatic prostate cancer who underwent such treatment. PATIENTS AND METHODS: Patients with newly diagnosed metastatic prostate cancer who had adequate hematologic, hepatic, renal, neurologic, and coagulation parameters were treated by combined androgen deprivation and suramin plus hydrocortisone. Suramin was administered on a 78-day fixed dosing schedule (one cycle), and suramin treatment cycles were repeated every 6 months for a total of four cycles. The statistical design was developed on the basis of the feasibility of administering suramin, as judged by the number of patients who developed neurotoxicity of grade 3 or higher or by treatment interruption of 4 weeks or longer due to any persistent suramin-related toxicity. RESULTS: Of the 62 patients enrolled onto the study between August 1994 and January 1997, 59 were eligible and assessable for toxicity on the first cycle. Thirty-two (54%) of 59 patients received a second cycle, 13 (22%) of 59 patients received a third cycle, and only five patients (8%) received a fourth cycle. During the first cycle, 27 patients were removed from the study: 17 because of toxicity, five because of disease progression, two who had died, and three because of other reasons. There was one therapy-related death. Grade 4 toxicities were noted in 11 and three patients during first and second courses, respectively. Neurotoxicity of grade 3 or higher was observed in nine and seven patients during the first and second cycles, respectively. Fifteen patients had treatment interruptions of 4 weeks or longer. Overall, only 54% (95% confidence interval, 41% to 67%) of the patients demonstrated acceptable limits of toxicity. CONCLUSION: Suramin plus hydrocortisone and androgen deprivation has limited applicability in the treatment of patients with newly diagnosed metastatic prostate cancer. PMID- 10694556 TI - Neoadjuvant chemotherapy and hormonal therapy followed by radical prostatectomy: feasibility and preliminary results. AB - PURPOSE: We assessed the feasibility and efficacy of integrating chemotherapy and androgen ablation with radical prostatectomy in patients with locally advanced prostate cancer. The neoadjuvant approach was adopted because it allows an in situ assessment of antitumoral activity. PATIENTS AND METHODS: Thirty-three patients were enrolled who met the clinical criteria of stage T1-2, Gleason score of >/= 8 or T2b-T2c, Gleason score of 7 and prostate-specific antigen (PSA) level greater than 10 ng/mL (n = 15), or clinical stage T3 (n = 18). Therapy consisted of 12 weeks of ketoconazole and doxorubicin alternating with vinblastine, estramustine, and androgen ablation followed by prostatectomy. The ability of neoadjuvant chemotherapy and hormonal therapy to induce a 20% rate of pT0 in the prostatectomy specimen as well as surgical feasibility were assessed. RESULTS: Chemotherapy complications were comparable to those reported with this regimen previously. No major intraoperative complications occurred. Postoperative complications occurred in 10 (33%) of 30 patients. One patient died at home after discharge (postoperative day 17; no autopsy was performed). Ten (33%) of the 30 patients had organ-confined disease, and 20 (70%) of 30 had extraprostatic extension; 11 (37%) of the 30 had positive lymph nodes. Only five (17%) of 30 exhibited positive surgical margins. All patients achieved an undetectable PSA level postoperatively, and 20 of the surviving 29 patients remain without disease recurrence with a median follow-up of 13 months (range, 9 to 18 months). CONCLUSION: Chemotherapy and androgen ablation followed by radical prostatectomy was feasible in patients with locally advanced prostate cancer. Although the goal of achieving a 20% rate for pT0 status was not achieved, we believe this type of integrated therapeutic strategy should be investigated further for its ability to alter the course of regionally advanced prostate cancer. PMID- 10694557 TI - Phase II study of cisplatin and paclitaxel in advanced carcinoma of the urothelium: an Eastern Cooperative Oncology Group Study. AB - PURPOSE: Cisplatin and paclitaxel are active agents in advanced urothelial cancer. A phase II trial of this combination was performed to determine the activity and toxicity of these agents in a multi-institutional setting. PATIENTS AND METHODS: Fifty-two patients with advanced urothelial carcinoma were treated on one day with paclitaxel 175 mg/m(2) over 3 hours followed by cisplatin 75 mg/m(2), both intravenously, every 21 days. Cycles were repeated every 21 days until progression or a maximum of six cycles. RESULTS: Twenty-six patients obtained an objective response, for an overall response rate of 50% (95% confidence interval, 36% to 64%). Four patients achieved complete clinical responses. The median overall survival time for the group was 10.6 months. Toxicity was moderate, with granulocytopenia and neurotoxicity being the most common side effects noted. CONCLUSION: The combination of cisplatin and paclitaxel is active in advanced urothelial cancer. Responses in visceral, nodal, and soft tissues sites were observed. Granulocytopenia without fever and grade 2/3 neurotoxicity were common. The confidence interval of the overall response rate in this study overlaps most of the other reported regimens. The optimal therapy for advanced urothelial cancer remains undefined. PMID- 10694558 TI - Topotecan has substantial antitumor activity as first-line salvage therapy in platinum-sensitive epithelial ovarian carcinoma: A Gynecologic Oncology Group Study. AB - PURPOSE: Topotecan is known to be active in recurrent ovarian cancer, but most prior studies have focused on platinum-resistant or refractory populations. This study was undertaken to define the response rate and progression-free interval in platinum-sensitive patients. PATIENTS AND METHODS: Patients with recurrent ovarian cancer after one or two prior chemotherapy regimens and in whom the interval between prior platinum therapy and the initiation of protocol therapy was greater than 6 months were treated with topotecan 1.5 mg/m(2) intravenously over 30 minutes daily for 5 days, with this cycle repeated every 21 days. RESULTS: Forty-eight patients were entered onto the study; 47 were assessable for toxicity and 46 for response. The response rate was 33% (two complete responses and 13 partial responses), with a median response duration of 11.2 months. Hematologic toxicity predominated but was manageable in most patients with frequent incorporation of cytokines and RBC and platelet transfusions. Fatigue was reported in 15 patients and resulted in the discontinuation of therapy in five responding patients. CONCLUSION: Topotecan is an active drug in platinum sensitive ovarian cancer, with significant but manageable hematologic toxicity. Fatigue is also a common problem that may be dose-limiting in some patients. PMID- 10694559 TI - Evaluation of soy phytoestrogens for the treatment of hot flashes in breast cancer survivors: A North Central Cancer Treatment Group Trial. AB - PURPOSE: Hot flashes represent a significant clinical problem for some breast cancer survivors. Safe, effective treatment is needed for this prominent clinical problem. Although it has been shown that estrogen or progesterone replacement therapy can alleviate this problem, there are continued safety concerns regarding the use of hormonal therapies in these women. Based on anecdotal information, we hypothesized that soy-derived phytoestrogens, weak estrogen-like substances in the soybean that demonstrate estrogen agonist and/or antagonist effects when they bind to estrogen receptors, could alleviate hot flashes. This current trial was designed to investigate this hypothesis. PATIENTS AND METHODS: This double-blind clinical trial involved breast cancer survivors with substantial hot flashes. After randomization, patients underwent a 1-week baseline period with no therapy. This was followed by 4 weeks of either soy tablets or placebo. The patients then crossed over to the opposite arm in a double-blind manner for the last 4 weeks. Patients completed a daily questionnaire documenting hot flash frequency, intensity, and perceived side effects. RESULTS: Of the 177 women who were randomized and started the study substance, 155 (88%) provided useable data over the first 5 weeks; 149 provided usable data over the entire 9 weeks. There was no suggestion that the soy product was more effective in reducing hot flashes than the placebo. At study completion, patients preferred the soy product 33% of the time, the placebo 37% of the time, and neither substance 31% of the time. No toxicity was observed. CONCLUSION: The soy product did not alleviate hot flashes in breast cancer survivors. PMID- 10694560 TI - High local recurrence risk after breast-conserving therapy in node-negative premenopausal breast cancer patients is greatly reduced by one course of perioperative chemotherapy: A European Organization for Research and Treatment of Cancer Breast Cancer Cooperative Group Study. AB - PURPOSE: Patients with invasive breast cancer may develop a local recurrence (LR) after breast-conserving therapy (BCT). Younger age has been found to be an independent risk factor for LR. Within a group of premenopausal node-negative breast cancer patients, we studied risk factors for LR and the effect of perioperative chemotherapy (PeCT) on LR. PATIENTS AND METHODS: The European Organization for Research and Treatment of Cancer (EORTC) conducted a randomized trial (EORTC 10854) to compare surgery followed by one course of PeCT (fluorouracil, doxorubicin, and cyclophosphamide) with surgery alone. From patients treated on this trial, we selected premenopausal patients with node negative breast cancer who were treated with BCT to examine whether histologic characteristics and the expression of various proteins (estrogen receptor, progesterone receptor, p53, Ki-67, bcl-2, CD31, c-erbB-2/neu) are risk factors for subsequent LR. Also, the effect of one course of PeCT on the LR risk (LRR) was studied. RESULTS: Using multivariate analysis, age younger than 43 years (relative risk [RR], 2.75; 95% confidence interval [CI], 1.46 to 5.18; P =.002), multifocal growth (RR, 3.34; 95% CI, 1.27 to 8.77; P =.014), and elevated levels of p53 (RR, 2. 14; 95% CI, 1.13 to 4.05; P =.02) were associated with higher LRR. Also, PeCT was found to reduce LRR by more than 50% (RR, 0.47; 95% CI, 0.25 to 0.86; P =.02). Patients younger than 43 years who received PeCT achieved similar LR rates as those of patients younger than 43 years who were treated with BCT alone. CONCLUSION: In premenopausal node-negative patients, age younger than 43 years is the most important risk factor for LR after BCT; this risk is greatly reduced by one course of PeCT. The main reason for administering systemic adjuvant treatment is to improve overall survival. The important reduction of LR after BCT is an additional reason for considering systemic treatment in young node-negative patients with breast cancer. PMID- 10694561 TI - Trauma history as a predictor of psychologic symptoms in women with breast cancer. AB - PURPOSE: To identify predictors of psychiatric problems in women with early-stage breast cancer. PATIENTS AND METHODS: One hundred sixty women with early-stage breast cancer were recruited from three treatment centers. They filled out self report questionnaires, including a medical history and demographic survey, the Trauma History Questionnaire, Life Event Questionnaire, Brief Symptom Inventory, Beck Depression Inventory, and Duke-UNC Functional Social Support Questionnaire, and were evaluated using the Structured Clinical Interview for DSM-III-R. RESULTS: Hierarchical regression analyses indicated that four of five variable sets made a significant incremental contribution to outcome prediction, with 35% to 37% of the variance explained. Outcomes were predicted by demographic variables, trauma history variables, precancer psychiatric diagnosis, recent life events, and perceived social support. Cancer treatment variables did not predict outcome. CONCLUSION: The findings highlight the important roles of trauma history and recent life events in adjustment to cancer and have implications for screening and treatment. PMID- 10694562 TI - Long-term prognosis after resection of hepatocellular carcinoma associated with hepatitis B-related cirrhosis. AB - PURPOSE: The optimum management of hepatocellular carcinoma (HCC) associated with cirrhosis has not yet been clarified. Very few data are available in the literature regarding the prognosis after resection of HCC associated with hepatitis B virus (HBV)-related cirrhosis. This study evaluated the long-term results and prognostic factors after resection of HCC complicating HBV-related cirrhosis. PATIENTS AND METHODS: One hundred forty-six patients with HBV-related Child's A or B cirrhosis who had undergone resection of HCC over a 10-year period were prospectively studied for long-term results. They were compared with 155 noncirrhotic patients with HBV-related HCC resected in the same period. RESULTS: The overall survival results of cirrhotic patients after resection of HCC were comparable to those of noncirrhotic patients (5-year survival, 44.3% v 45.6%, respectively; P =.216), but the former group had significantly smaller tumors. Stratified according to tumor size, the survival results were similar between cirrhotic and noncirrhotic patients with tumors epsilon3) for Alzheimer's disease (AD). ApoE may affect AD pathogenesis by promoting deposition of the amyloid-beta (Abeta) peptide and its conversion to a fibrillar form. To determine the effect of apoE on Abeta deposition and AD pathology, we compared APP(V717F) transgenic (TG) mice expressing mouse, human, or no apoE (apoE(-/-)). A severe, plaque-associated neuritic dystrophy developed in APP(V717F) TG mice expressing mouse or human apoE. Though significant levels of Abeta deposition also occurred in APP(V717F) TG, apoE(-/-) mice, neuritic degeneration was virtually absent. Expression of apoE3 and apoE4 in APP(V717F) TG, apoE(-/-) mice resulted in fibrillar Abeta deposits and neuritic plaques by 15 months of age and substantially (>10-fold) more fibrillar deposits were observed in apoE4-expressing APP(V717F) TG mice. Our data demonstrate a critical and isoform-specific role for apoE in neuritic plaque formation, a pathological hallmark of AD. PMID- 10694578 TI - HLA B*5701 is highly associated with restriction of virus replication in a subgroup of HIV-infected long term nonprogressors. AB - A unique cohort of HIV-1-infected long term nonprogressors (LTNP) with normal CD4(+) T cell counts and <50 copies/ml of plasma were prospectively recruited for study. HLA typing revealed a dramatic association between the HLA B*5701 class I allele and nonprogressive infection [85% (11 of 13) vs. 9.5% (19 of 200) in progressors; P < 0. 001]. Antigen-specific CD8(+) T cells were enumerated by flow cytometric detection of intracellular IFN-gamma in response to HIV antigens and HLA B*57-gag tetramer staining. No quantitative differences in the total HIV specific CD8(+) T cell responses were observed between B*57(+) LTNP and five B*57(+) progressors (P = 0.4). Although similar frequencies of peptide specific CD8(+) T cells were also found, the gag-specific CD8(+) T cell response in the LTNP group was highly focused on peptides previously shown to be B*57-restricted. These findings indicate that, within this phenotypically and genotypically distinct cohort, a host immune factor is highly associated with restriction of virus replication and nonprogressive disease. They also strongly suggest a mechanism of virus specific immunity that directly operates through the B*5701 molecule. Further characterization of qualitative differences in the virus specific responses that distinguish HLA B*57(+) LTNP from progressors may ultimately define mechanisms of effective immune mediated restriction of virus replication. PMID- 10694579 TI - The interaction of nitric oxide (NO) with the yeast transcription factor Ace1: A model system for NO-protein thiol interactions with implications to metal metabolism. AB - Nitric oxide (NO) was found to inhibit the copper-dependent induction of the yeast CUP1 gene. This effect is attributable to an inhibition of the copper responsive CUP1 transcriptional activator Ace1. A mechanism is proposed whereby the metal binding thiols of Ace1 are chemically modified via NO- and O(2) dependent chemistry, thereby diminishing the ability of Ace1 to bind and respond to copper. Moreover, it is proposed that demetallated Ace1 is proteolytically degraded in the cell, resulting in a prolonged inhibition of copper-dependent CUP1 induction. These findings indicate that NO may serve as a disrupter of yeast copper metabolism. More importantly, considering the similarity of Ace1 to other mammalian metal-binding proteins, this work lends support to the hypothesis that NO may regulate/disrupt metal homeostasis under both normal physiological and pathophysiological circumstances. PMID- 10694581 TI - Springtime of hope. PMID- 10694580 TI - In vivo natural killer cell activities revealed by natural killer cell-deficient mice. AB - Studies of natural killer (NK) cell function in vivo have been challenging primarily due to the lack of animal models in which NK cells are genetically and selectively deficient. Here, we describe a transgenic mouse with defective natural killing and selective deficiency in NK1.1(+) CD3(-) cells. Despite functionally normal B, T, and NK/T cells, transgenic mice displayed impaired acute in vivo rejection of tumor cells. Adoptive transfer experiments confirmed that NK1.1(+) CD3(-) cells were responsible for acute tumor rejection, establishing the relationship of NK1.1(+) CD3(-) cells to NK cells. Additional studies provided evidence that (i) NK cells play an important role in suppressing tumor metastasis and outgrowth; (ii) NK cells are major producers of IFNgamma in response to bacterial endotoxin but not to interleukin-12, and; (iii) NK cells are not essential for humoral responses to T cell-independent type 2 antigen or the generalized Shwartzman reaction, both of which were previously proposed to involve NK cells. PMID- 10694583 TI - Ocean tides. PMID- 10694582 TI - The threat of stress to the heart health of Canadians: results of the Foundation health survey. PMID- 10694584 TI - Hypertrophic cardiomyopathy in pediatric patients: effect of verapamil on regional and global left ventricular diastolic function. AB - OBJECTIVE: To assess the effects of treatment with verapamil on regional and global left ventricular (LV) diastolic function in paediatric patients with hypertrophic cardiomyopathy (HCM). DESIGN: Twelve patients (age range 5.1 to 12.3 years, median 8.6) with HCM were evaluated during ongoing chronic oral treatment with verapamil (4 mg/kg/day) and four days after withdrawal of therapy. Twelve age- and body surface area-matched normal children served as controls. In an echocardiographic study, global LV diastolic function was evaluated by assessing isovolumic relaxation time (IVRT) and mitral flow indexes, including peak filling rate normalized to mitral stroke volume (PFR/SV). In addition, regional LV diastolic function was assessed by pulsed-wave Doppler tissue imaging at the subendocardial portion of the middle region of the anterior and posterior interventricular septum, and anterolateral and inferior walls to measure the peak velocities and the velocity-time integrals of myocardial excursion in both early diastole and atrial systole. In addition, as an index of diastolic asynchrony (AsyI), the variation in time to peak filling rate, measured as the time from the peak of the R wave on the electrocardiogram to the peak of the regional E wave, among the four myocardial regions was defined by subtracting the smallest value from the greatest and expressing the difference as a percentage of the smallest value. RESULTS: Compared with the controls, patients with HCM without therapy showed a longer IVRT (P<0.01) and a decrease in PFR/SV (P<0.01) without a compensatory increase in filling during atrial systole. Oral administration of verapamil induced a significant shortening of the IVRT (P=0.003) and an increase in PFR/SV (P=0.02). Furthermore, patients with HCM without therapy showed a significantly longer time to peak filling rate (P<0.01) associated with a decreased peak velocity in early diastole without a concomitant increase in peak velocity during atrial systole in each of the myocardial regions. Furthermore, the AsyI was higher in the HCM group than in controls (19% versus 6%, respectively), and this index was inversely correlated with the PFR/SV (r=-0.86, P<0.001). The regional diastolic velocity of the myocardium at each of the four analyzed regions was not significantly different with verapamil, but the AsyI was significantly lower (P<0.05). CONCLUSIONS: Children with HCM show abnormalities of both global and regional LV diastolic function. In these patients, chronic administration of verapamil plays a crucial role in the improvement in global LV filling and, as a consequence, in clinical manifestations. The beneficial effects of verapamil seem to be related to a reduction in diastolic asynchrony more than to significant changes in diastolic velocities of the myocardial fibres. PMID- 10694585 TI - Oral anticoagulant therapy for heart disease: results in actual cardiology practice. AB - OBJECTIVE: To determine whether the success and complication rates of oral anticoagulation obtained in large, well controlled trials, upon which recommendations are based, are comparable with routine cardiology practice. DESIGN: An observational, prospective cohort study collected data on all patients followed at an anticoagulant clinic over one calendar year. PATIENTS: One thousand and seventy-eight patients anticoagulated for cardiovascular indications, mainly atrial fibrillation, prosthetic valves and ventricular dysfunction, were followed for 804 patient-years of treatment. No patient was lost to follow-up. INTERVENTIONS: Telephone conversations and regular verification of medical files were used to record and classify all bleeding and thromboembolic events according to severity. International normalized ratios (INR) were compared with target ranges. RESULTS: One hundred and twelve bleeding events, ie, 13.9/100 patient-years (% p-y), were recorded, of which 61 required medical attention. Major hemorrhages, defined as those requiring treatment or hospital observation for more than 24 h, occurred in 15 instances (1.9% p-y). Among these, 9 (1.1% p-y) were classified as life threatening, with four being fatal (0,5% p-y). Twenty-two thromboembolic events (2.7% p-y) occurred, of which 10 were major (1.2% p-y), leaving three patients (0.4% p-y) with long term sequelae and causing two deaths (0.25% p-y). INRs were within target range 62.3% of the time, with 2.2% of values recorded above 5 and 0.3% above 10. CONCLUSION: The low failure and complication rates obtained in large, controlled trials are similar to those observed in actual cardiology practice. PMID- 10694586 TI - [Myocardial infarction with normal or near normal coronary arteries: late outcome of seven patients]. AB - Myocardial infarction with normal or near normal coronary arteries: Late outcome of seven patients Myocardial infarction with normal or near normal coronary arteries probably results from thrombotic occlusion that is followed by lysis and recanalization; in about a third of affected patients, coronary spasm appears to be the trigger. The clinical profile of this syndrome is well known: patients are young and without coronary risk factors except cigarette smoking, and they have no angina before or after their infarction. However, the late prognosis remains controversial. Between December 1976 and April 1984, myocardial infarction with normal or near normal coronary arteries was diagnosed in seven patients who were subsequently followed for a mean of 19 years. Observations during this follow-up show that the majority of patients experienced one or more recurrences that, in three cases, initiated the development of a severe ischemic cardiomyopathy. PMID- 10694587 TI - Double-chambered right ventricle in 73 patients: spectrum of the disease and surgical results of transatrial repair. AB - OBJECTIVE: To review the spectrum of double-chambered right ventricle (DCRV) and the outcome of surgical repair in patients diagnosed between February 1988 and March 1999. DESIGN: The charts of patients with DCRV were studied. SETTING: Tertiary care hospital. PATIENTS AND METHODS: A total of 73 patients were identified. Sixty-nine underwent surgical repair, while four are awaiting surgery. The repair was through a transatrial approach in 61 patients, while in eight an additional ventriculotomy was performed. MAIN RESULTS: An associated ventricular septal defect (VSD) was present in 56 of 73 patients (77%). These patients were significantly younger (P<0.05) than the 17 patients without a VSD. Among patients with a VSD, the 31 requiring patch closure were significantly younger than the 25 patients having direct closure. Five older patients among those with intact septum had impaired right ventricular (RV) function as well as higher intraventricular gradients. At surgery the intraventricular obstruction was relieved by myomectomy. There was no hospital or late mortality. Following surgery, at a mean follow-up of 13.6 months, no increase in the intraventricular gradient was detected by Doppler echocardiography. CONCLUSIONS: The development of DCRV is associated with VSD in early life. The probability of the presence of a VSD decreases with age. The disease is progressive, resulting in increased intracavitary gradient within the RV and in RV impairment if it is not treated in a timely fashion. Transatrial repair is safe with excellent midterm results. In the presence of high gradients within the RV, a ventriculotomy may be necessary to obtain acceptable results. PMID- 10694588 TI - Body surface mapping of retrograde P waves in the intact dog by simulation of accessory pathway re-entry. AB - OBJECTIVES: To investigate a noninvasive technique to localize the atrial insertion site of concealed accessory pathways based on the analysis of body surface potential maps (BSPMs) of retrograde P waves in dogs with simulated retrograde pathways. ANIMALS AND METHODS: Orthodromic tachycardias were simulated by atrial stimulations at eight different sites around the atrioventricular ring with long (250 ms and 300 ms) and short (100 ms and 130 ms) coupling times in 14 anesthetized dogs to have P waves well separated from the T wave or occurring during the T wave, respectively. The distance between pacing sites was 15 to 40 mm in group 1 (eight dogs) and 2 mm (in the right atrial free wall region) in group 2 (six dogs). Beats were signal-averaged during 30 s and BSPMs were constructed from 63 unipolar leads. RESULTS: The P wave BSPM pattern for any specific stimulation site was stable and reproducible (correlation coefficient greater than 0.98), and similar in different dogs at long coupling interval stimulations. The thoracic distribution of negative potentials and position of the potential minimum clearly identified the stimulation site when long coupling time stimulations were used. The spatial resolution of the technique as determined by comparison of correlation coefficients in group 2 was 6 mm (P<0.05). When short coupling time stimulations were used (fast tachycardia simulation), the T wave masked the P wave potential distribution in four of eight dogs, but the retrograde P wave map could still be accurately extracted by subtracting a straight line joining the onset and offset of the P wave in 24 of 28 (86%) of the tachycardia simulation sites in these four dogs. CONCLUSIONS: The BSPM patterns of simulated retrograde P waves are specifically related to the site of atrial stimulation. Although the T wave altered these BSPM patterns, a subtraction technique recovered the pattern of the retrograde P wave in 93% of all simulated orthodromic tachycardias. The spatial resolution of the retrograde P wave BSPM method was 6 mm. PMID- 10694589 TI - Infusion of an antialpha4 integrin antibody is associated with less neoadventitial formation after balloon injury of porcine coronary arteries. AB - BACKGROUND: The alpha4beta1 (or very late antigen-4 [VLA-4]) integrin is thought to play a role in inflammatory processes, mediating mononuclear leukocyte infiltration. The adventitial response to balloon injury is an important determinant of neointimal formation and arterial remodelling. OBJECTIVES: To determine whether the monoclonal antibody hHP1/2 directed against the human alpha4-integrin subunit decreases neoadventitial formation and subsequent luminal narrowing following balloon injury. DESIGN: Randomized, double-blind, placebo controlled study. SETTING: Tertiary care, Canadian university hospital vascular biology laboratory. ANIMALS AND METHODS: In 16 pigs, two coronary arteries were injured with an oversized balloon, while a third coronary artery was designated as an uninjured control vessel. One hour before balloon injury, 5 mg/kg of hHP1/2 was administered to eight animals, while another eight animals received an infusion of a saline placebo. Animals were killed three and 14 days following balloon injury. MAIN RESULTS: Administration of hHP1/2 resulted in an immediate decrease in circulating monocyte and lymphocyte counts. These parameters returned to normal within three days. There was a decrease in neoadventitial formation 14 days after arterial injury in pigs treated with hHP1/2 compared with controls (2.26+/-0.77 versus 3.42+/-1.01 mm, respectively, P=0.04). There was a loss of lumen area between days 3 (4.33+/-1.09 mm2) and 14 (3.09+/-0.38 mm2, P=0.02) after balloon injury in pigs treated with saline, but not in the pigs treated with hHP1/2. CONCLUSIONS: Administration of an antibody to the alpha4-integrin subunit is associated with less neoadventitial formation and less lumenal narrowing after balloon injury. This novel therapy may play an important role in modulating arterial remodelling and thereby may reduce restenosis following percutaneous coronary interventions in humans. PMID- 10694590 TI - Cardiac angiosarcoma: two cases and a review of the literature. AB - BACKGROUND: Primary cardiac tumours are rare, and primary malignant cardiac tumours even rarer. Of these, cardiac angiosarcomas are uncommon and, until recently, almost invariably diagnosed at the time of autopsy, primarily because the symptoms are initially nonspecific and do not become manifest until the tumour is advanced. METHODS: Two patients, who presented in quite different manners and were diagnosed at autopsy and at open surgical biopsy, are presented. The literature on cardiac angiosarcomas is reviewed critically, with emphasis on presentation and morphology. RESULTS: This review of the literature shows that, with increasing availability of newer diagnostic tools, especially noninvasive ones, diagnosis of this rare lesion can be made at an early stage and confirmed at cardiac biopsy or cardiac surgery. Unfortunately, so far, the results remain virtually uniformly poor, though occasionally survival at up to 53 months has been reported. PMID- 10694591 TI - Fatal myocardial embolus after myectomy. AB - Coronary embolism is an infrequent phenomenon. A 56-year-old man with hypertrophic obstructive cardiomyopathy and severe mitral regurgitation who underwent left ventricular septal myectomy and mitral valve annular repair is presented. The patient had a cardiac arrest 36 h after surgery. Cardiac standstill, tamponade and a left ventricular rupture were noted when the chest was opened during attempted resuscitation. Autopsy revealed an occlusive embolus of myocardium in the proximal left anterior descending coronary artery. It showed pathological features of hypertrophic cardiomyopathy. There was an extensive acute transmural anteroseptal left ventricular myocardial infarction with rupture of the anterior free wall. Embolism of myocardium - to the coronary arteries, the systemic circulation or the pulmonary circulation - is a rare event, with only nine other cases reported in the literature in the past 30 years. This is the first reported case of myocardial embolus to a coronary artery in a patient with hypertrophic obstructive cardiomyopathy following septal myectomy. PMID- 10694592 TI - An unusual case of ST elevation in a 39-year-old man. AB - A 39-year-old man presented to a university hospital emergency department with anginal chest pain, ventricular tachycardia and ST elevation in the anterolateral leads (V3 to V6, I and aVL). Due to discrepancies in the history and physical examination, thrombolysis was withheld until a past electrocardiogram could be obtained, which was unchanged. Subsequent investigations revealed no evidence of myocardial necrosis, and the patient was diagnosed with hypertrophic cardiomyopathy. This is the first reported case of hypertrophic cardiomyopathy with ST elevation as the predominant electrocardiographic abnormality. In patients with discrepancies in the clinical presentation, it is essential to obtain past elecrocardiograms to ensure appropriate utility of thrombolysis. PMID- 10694593 TI - Sirolimus, a new potent immunosuppressant agent for refractory cardiac transplantation rejection: two case reports. AB - Rejection remains a major cause of both early and late morbidity and mortality following heart transplantation despite major advances in immunosuppressive therapy. A major hurdle in the successful management of patients who have undergone heart transplantation is preventing and treating graft rejection. Cyclosporine, mycophenolate mofetil, azathioprine, tacrolimus (FK506) and OKT3 are well documented, effective immunosuppressive medications that prevent and treat acute or chronic rejection following heart transplantation. One of the macrolide antibiotics, sirolimus, is known to have immunosuppressant activity. In two patients with chronic rejection of cardiac grafts refractory to usual antirejection medications, sirolimus was successfully used to suppress graft rejection. Both patients continued to be rejection free after 10 months of sirolimus treatment despite significant decreases in doses of other immunosuppressants. PMID- 10694594 TI - The World Heart Federation's white book: impending global pandemic of cardiovascular diseases: challenges and opportunities for the prevention and control of cardiovascular diseases in developing countries and economies in transition. PMID- 10694596 TI - Donald l. Paulson (1912-1999) PMID- 10694597 TI - Editorial foreword PMID- 10694595 TI - Cardiovascular disease and the global village. PMID- 10694598 TI - The core of leadership. PMID- 10694599 TI - Chest wall resection for locally recurrent breast cancer: is it worthwhile? AB - OBJECTIVE: The effectiveness of chest wall resection for locally recurrent breast cancer as cancer treatment remains poorly defined, possibly because of the general impression that locally recurrent disease is a harbinger of rapidly progressive metastatic disease and that extensive surgical treatment in these patients is inappropriate. Reports to date have focused on technical feasibility, not long-term outcome. METHODS: We reviewed our experience with 38 women who underwent chest wall resection for locally recurrent breast cancer between October 1987 and May 1997. Overall survival was calculated by the Kaplan-Meier method and the significance of prognostic variables evaluated by log-rank and Cox regression analyses. RESULTS: The operative mortality rate was 0%. Overall survival at 1, 3, and 5 years after chest wall resection was 74%, 41%, and 18%, respectively, and the proportion of patients free of local recurrence at 1, 3, and 5 years was 59%, 42%, and 13%, respectively. Regional nodal disease and size of largest tumor nodule (>4 cm) were significant predictors of local re recurrence (P <.01, P =.04); lymph node metastasis was the only predictor of long term survival (P <.01). Patients with and without synchronous sites of metastatic disease had near-identical 3-year survivals. CONCLUSIONS: Chest wall resection for locally recurrent breast cancer has a low mortality. However, a significant number of patients have the development of local re-recurrence or metastases, and 5-year survival is limited. It is unlikely that complete resection of all locally recurrent disease improves survival. Future studies should focus on the quality of palliation achieved. PMID- 10694600 TI - Induction chemotherapy before surgery for early-stage lung cancer: A novel approach. Bimodality Lung Oncology Team. AB - OBJECTIVE: This phase II trial assessed the feasibility, as measured by response rate, toxicity, resectability rate, and surgical morbidity and mortality rates, of perioperative paclitaxel and carboplatin chemotherapy in patients with early stage non-small cell lung carcinoma. METHODS: All patients required negative mediastinoscopy results and adequate medical parameters to undergo induction chemotherapy and an operation. Superior sulcus patients were excluded. Chemotherapy consisted of paclitaxel 225 mg/m(2) over 3 hours and carboplatin (area under the curve = 6) every 21 days for 2 cycles preoperatively. Three postoperative cycles of chemotherapy were planned for patients undergoing complete resection. RESULTS: Between June 1996 and July 1998, 94 patients were entered into the study. Sixty-five (69%) were men, and the median age was 64 years (range, 34-79 years). After induction chemotherapy, 53 of 94 (56%; 95% confidence interval, 46%-67%) had a major objective response, 88 (94%) underwent surgical exploration, and 81 (86%; 95% confidence interval, 78%-92%) underwent complete resection. Reasons for not undergoing an operation included disease progression (n = 3), clinically unresectable status (n = 1), death (n = 1), and patient lost to follow-up (n = 1). Two postoperative deaths occurred. Six (6%; 95% confidence interval, 0%-13%) pathologic complete responses were observed. Ninety (96%) patients received the planned preoperative chemotherapy versus 45% receiving postoperative chemotherapy. No unexpected chemotherapy or surgical morbidity occurred. The 1-year survival is currently estimated at 85%, and the median survival has not yet been reached. CONCLUSIONS: Induction chemotherapy with paclitaxel and carboplatin is feasible and produces a high response rate with acceptable morbidity and mortality rates in early-stage non-small cell lung carcinoma. A prospective randomized trial comparing 3 cycles of induction chemotherapy and surgery with surgery alone in early-stage non-small cell lung carcinoma is planned. PMID- 10694601 TI - Does pneumonectomy for lung cancer adversely influence long-term survival? AB - OBJECTIVE: The increased operative mortality associated with pneumonectomy has stimulated the use of lung-sparing operations such as sleeve lobectomy. Whether pneumonectomy adversely affects long-term outcome after lung resection is unknown. METHODS: We reviewed the cases of patients who underwent lobectomy/bilobectomy or pneumonectomy because of non-small cell lung cancer between January 1980 and June 1998. Survival curves were compared by the log-rank test. Covariates were determined for operative mortality and survival using logistic regression analysis and Cox proportional hazards estimation, respectively. RESULTS: There were 259 men and 183 women who underwent lobectomy/bilobectomy (340) or pneumonectomy (102). Operative mortality was 36 (8.1%) patients overall, 24 (7.0%) for lobectomy/bilobectomy and 12 (12%) for pneumonectomy. Mean follow-up was 41 months (range 0-222 months). Median survival was worse for pneumonectomy (stage II: 17.9 vs 36.3 months, log-rank P =. 05; stage III: 11.7 vs 21.3 months, log-rank P =.07). However, important covariates for survival were age, primary tumor status, regional nodal status, and forced expiratory volume in 1 second. After adjusting for these covariates, survival did not differ significantly between the types of operations (hazard ratio for pneumonectomy 1.21; 95% CI 0.88-1.68). CONCLUSIONS: We did not detect a significant long-term adverse influence of pneumonectomy on survival after adjusting for other prognostic factors, but randomized clinical trials would be needed to definitively address this issue. PMID- 10694602 TI - Preemptive gastrointestinal tract management reduces aspiration and respiratory failure after thoracic operations. AB - OBJECTIVES: Respiratory failure is the major mode of death after general thoracic operations. However, respiratory failure may develop from two very different mechanisms: aspiration, often caused by ileus, and pneumonia, which often results from poor pain control. Epidural catheters help control pain and prevent pneumonia but contribute to ileus and may increase aspiration. We report a decrease in the incidence of aspiration after changing postoperative care to include gastrointestinal tract management. METHODS: All patients undergoing elective thoracotomy by a single surgeon were evaluated for hospital mortality and morbidity. For the first 21 months, patients did not receive an intraoperative nasogastric tube and were prescribed an "advance as tolerated" diet after the operation (n = 125). For the second period, nasogastric tubes were placed intraoperatively and patients received nothing by mouth the day of operation, clear liquids the first day, and a regular diet the second day (n = 153). Pneumonia was considered to have developed if infiltrates developed in a single lobe or two adjoining lobes and culture of the sputa grew a dominant organism. Patients were considered to have aspirated if diffuse infiltrates developed or cultures grew multiple organisms. Significance of results was determined by chi(2) testing. RESULTS: A total of 278 patients underwent elective lung resection over a 3(1/2)-year period, 125 with ad libitum dietary management and 153 with intensive management of the gastrointestinal tract. Six patients (4.84%) aspirated before the institution of gastrointestinal tract management, whereas none (0.0%) aspirated after the change. This difference was significant (P =.01). Respiratory mortality was eliminated in the group with gastrointestinal tract management (P =.04). CONCLUSIONS: Aspiration and its subsequent respiratory failure and mortality can be decreased with preemptive gastrointestinal tract management. PMID- 10694603 TI - Chylothorax complicating esophagectomy for cancer: a plea for early thoracic duct ligation. AB - OBJECTIVE: Postoperative chylothorax remains an uncommon but potentially life threatening complication of esophagectomy for cancer, and the ideal management is still controversial. The aim of the study was to compare the outcomes of patients treated nonoperatively with those of patients promptly undergoing reoperation. METHODS: From 1980 to 1998, 1787 esophagectomies for esophageal or cardia cancer were performed, and 19 (1.1%) patients had postoperative chylothorax. We analyzed type of operation, surgical approach, delay of diagnosis of chylothorax, daily chest tube output, type of management, major complications, death, hospital stay, and final outcome. RESULTS: Of the 19 patients with chylothorax, 11 were initially managed nonoperatively (group A): 4 (36%) patients had spontaneous resolution of chylothorax, and the other 7 required reoperation for the persistence of a high-volume output. There were three infectious complications and one postoperative death in this group. No reliable predictive criteria of successful versus unsuccessful nonoperative management could be found. The 8 most recent patients underwent early reoperation (group B). All patients recovered, and no major complications possibly related to chylothorax or hospital deaths were observed. They were discharged after a median of 22 days (range, 12-85 days) compared with a median of 36 days (range, 21-64 days) for patients of group A. CONCLUSIONS: Early thoracic duct ligation is the treatment of choice for chylothorax occurring after esophagectomy. Reoperation should be performed immediately after the diagnosis is made to avoid the complications related to nutritional and immunologic depletion caused by prolonged nonoperative treatment. PMID- 10694604 TI - Secondary pulmonary hypertension does not adversely affect outcome after single lung transplantation. AB - OBJECTIVE: Primary and secondary pulmonary hypertension have been associated with poor outcomes after single lung transplantation. Some groups advocate double lung transplantation and the routine use of cardiopulmonary bypass during transplantation in this population. However, the optimal procedure for these patients remains controversial. The goal of our study was to determine the safety of single lung transplantation without cardiopulmonary bypass in patients with secondary pulmonary hypertension. METHODS: We retrospectively reviewed 76 consecutive patients with pulmonary parenchymal disease who underwent single lung transplantation from 1992 to 1998. Recipients were stratified according to preoperative mean pulmonary artery pressure. Secondary pulmonary hypertension was defined as parenchymal lung disease with a preoperative mean pulmonary artery pressure of 30 mm Hg or more. Patients with primary pulmonary hypertension or Eisenmenger's syndrome were excluded from analysis. RESULTS: Eighteen of 76 patients had secondary pulmonary hypertension. No patient with secondary pulmonary hypertension required cardiopulmonary bypass, whereas 1 patient without pulmonary hypertension required bypass. After the operation, no significant differences were seen in lung injury as measured by chest radiograph score and PaO(2)/FIO(2) ratio, the requirement for inhaled nitric oxide, the length of mechanical ventilation, the intensive care unit or hospital length of stay, and 30-day survival. There were no differences in the forced expiratory volume in 1 second or 6-minute walk at 1 year, or the incidence of rejection, infection, or bronchiolitis obliterans syndrome greater than grade 2. Survival at 1, 2, and 4 years after transplantation was 86%, 79%, and 65%, respectively, in the low pulmonary artery pressure group and 81%, 81%, and 61%, respectively, in the group with secondary pulmonary hypertension (P >.2). CONCLUSION: We found that patients with pulmonary parenchymal disease and concomitant secondary pulmonary hypertension had successful outcomes as measured by early and late allograft function and appear to have acceptable long-term survival after single lung transplantation. Our results do not support the routine use of cardiopulmonary bypass or double lung transplantation for patients with this disorder. PMID- 10694605 TI - Heart-lung versus double-lung transplantation for suppurative lung disease. AB - OBJECTIVE: The purpose of this study was to compare outcomes after heart-lung or double-lung transplantation in patients undergoing transplantation because of end stage suppurative lung disease. METHODS: We reviewed our experience in patients with cystic fibrosis or bronchiectasis who had heart-lung or double-lung transplantation between January 1988 and September 1997. Twenty-three patients (14 male, 21 cystic fibrosis) had heart-lung transplantation and 24 patients (8 male, 19 cystic fibrosis) had double-lung transplantation. There were no statistically significant differences between the groups in age, weight, preoperative creatinine level, cytomegalovirus status, maintenance immunosuppression, or donor demographics. Patients received induction therapy with monoclonal (OKT3) or polyclonal (rabbit anti-thymocyte globulin) antibody. RESULTS: Sixteen of 24 patients had double-lung transplantation after 1994 whereas 13 of 22 patients had heart-lung transplantation before 1991, allowing longer follow-up for the heart-lung group. Mean waiting times for transplantation were 270 +/- 245 days (heart-lung) and 361 +/- 229 days (double-lung; P =.20). The 1-, 3-, and 5-year actuarial survival figures were respectively 86%, 82%, and 65% (heart-lung) and 96%, 75%, and unavailable (double-lung; P = no significant difference). The 1-, 3-, and 5-year rates of freedom from obliterative bronchiolitis were respectively 77%, 61%, and 45% (heart-lung) and 86%, 78%, and unavailable (double-lung; P = no significant difference). Linearized overall infection rates (events/100 patient-days) were 2.05 +/- 0.33 (heart-lung) and 2.34 +/- 0.34 (double-lung; P = NS) at 3 months. Thirty-day survival was 100% (heart-lung) and 96% (double-lung). There were 7 late deaths among heart-lung recipients (3 obliterative bronchiolitis, 2 infection, 0 graft coronary artery disease, 2 other) whereas 2 late deaths related to obliterative bronchiolitis occurred in double-lung recipients. Graft coronary artery disease (all stenoses < 50%) affected 15% of heart-lung survivors, whereas 3 double-lung recipients (12.5%) required either bronchial dilatation or stenting. CONCLUSION: Heart-lung and double-lung transplantation provide similar palliation for patients with end stage suppurative lung disease. Therefore double-lung transplantation should be the preferred operation for most patients with end-stage suppurative lung disease. PMID- 10694606 TI - Evidence against a pivotal role of preformed antibodies in delayed rejection of a guinea pig-to-rat heart xenograft. AB - INTRODUCTION: Whereas the involvement of elicited xenoantibodies in delayed xenograft rejection is currently being substantiated, this study focuses on the role of the preformed fraction of xenoantibodies. METHODS: To check the influence of the latter, we combined pretransplant complement inactivation (cobra venom factor) and antibody reduction (plasmapheresis) in a guinea pig-to-rat heart transplant model. RESULTS: Antibody reduction on plasmapheresis before xenografting did not prolong delayed xenorejection in decomplemented rats, although the immunohistologic pattern lacked the immunoglobulin deposits along endothelial walls found in xenografts of merely decomplemented recipients. Astonishingly, plasmapheresis, if carried out 2 days before transplantation, almost tripled xenograft survival, although preformed antibody levels were completely restored and even rebounding at the time of grafting. The pattern and number of infiltrating cells did not differ in dependence of the timing of plasmapheresis nor did the proliferative response of lymphocytes in the mixed lymphocyte reaction differ. However, plasmapheresis led to a retarded decrease of the mononuclear cell tumor necrosis factor alpha secretory potential, which correlated well with a diminished immunohistologic staining of tumor necrosis factor alpha secreted by graft-infiltrating mononuclear cells. CONCLUSION: These findings argue against a pivotal role of preformed xenoantibodies in the pathomechanistic process of delayed xenograft rejection and challenge the therapeutic strategy to reduce preformed xenoantibody levels before xenotransplantation in complement-inactivated recipients. PMID- 10694607 TI - Raffinose improves the function of rat pulmonary grafts stored for twenty-four hours in low-potassium dextran solution. AB - OBJECTIVES: The perfect strategy for pulmonary graft preservation remains elusive. Experimental work supports the use of perfusates, such as Euro-Collins, University of Wisconsin, and low-potassium dextran solutions. We use low potassium dextran solution in our clinical program, but we aim for continued improvement. The trisaccharide raffinose has been shown to be responsible for the efficacy of University of Wisconsin perfusate in lung preservation. Raffinose is superior to a variety of other saccharides for this purpose. We tested the hypothesis that the addition of raffinose to low-potassium dextran solution might further improve graft function. METHODS: In a randomized blinded study with a rat left lung transplant model, donor lungs were flushed with either standard low potassium dextran solution or low-potassium dextran solution modified by the addition of 30 mmol/L raffinose (n = 5 for each group). Alprostadil (prostaglandin E(1), 500 microg/L) was added to the perfusates in accordance with our clinical practice. Grafts were stored inflated at 4 degrees C for 24 hours. After transplantation, recipients were ventilated with a fraction of inspired oxygen of 1 and a positive end-expiratory pressure of 2 cm H(2)O. Graft function was evaluated by measuring oxygenation at 2 hours after graft reperfusion, peak airway pressure throughout the reperfusion period, and the wet/dry lung weight ratio. RESULTS: The group receiving low-potassium dextran solution with raffinose demonstrated significantly higher oxygenation (oxygen tension, 370 +/- 45 mm Hg vs 150 +/- 64 mm Hg; P =.0025), lower peak airway pressures at 2 hours after lung reperfusion (11 +/- 2.7 mm Hg vs 16 +/- 2.4 mm Hg; P <.001), and a lower wet/dry weight ratio (4.7 +/- 1.26 vs 11 +/- 5. 0; P =.017). CONCLUSION: Modification of low-potassium dextran solution with the trisaccharide raffinose resulted in a significant improvement in graft function in this model and merits further evaluation with respect to the mechanisms involved. PMID- 10694608 TI - Highly efficient ex vivo gene transfer to the transplanted heart by means of hypothermic perfusion with a low dose of adenoviral vector. AB - BACKGROUND: Hypothermic conditions required for donor heart preservation may reduce gene-transfer efficiency. Experiments were designed to determine whether a perfusion technique could improve the efficiency of gene transfer to donor hearts. METHODS: An adenoviral vector encoding beta-galactosidase (3.5 x 10(8) plaque-forming units) was infused into explanted rat hearts under 4 conditions (each n = 6): (1) the virus was diluted in 350 microL of University of Wisconsin solution and infused as a high-pressure bolus into the coronary arteries of donor hearts through the aortic root; (2) the virus was diluted in 5 mL of University of Wisconsin solution and circulated by means of a peristaltic pump (flow, 0.75 mL/min) through the vasculature of the donor heart for 30 minutes; (3) 5 mL of viral solution was circulated as for group 2 for 15 minutes; and (4) 5 mL of viral solution was circulated for 5 minutes at a flow rate of 2.4 mL/min. Transduced hearts were transplanted into the abdomen of syngeneic rats, and transgene expression was assessed by means of immunoassay 4 days later. RESULTS: The median beta-galactosidase content was (1) 45.0 ng/mg protein (25th-75th percentile, 33-73 ng/mg), (2) 640 ng/mg protein (25th-75th percentile, 614-878 ng/mg), (3) 493.8 ng/mg protein (25th-75th percentile, 456-527 ng/mg), and (4) 503.3 ng/mg protein (25th-75th percentile, 475-562 ng/mg; P <.01 for group 2 vs group 1, and P <.05 for groups 3 and 4 vs group 1). Transgene expression was predominantly in myocytes and favored the subepicardial region of the right ventricle. CONCLUSION: Hypothermic perfusion of the donor heart with an adenoviral vector resulted in efficient transgene expression compared with that induced by a single bolus injection. PMID- 10694610 TI - Commentary PMID- 10694609 TI - Pulmonary function after modified venovenous ultrafiltration in infants: a prospective, randomized trial. AB - OBJECTIVE: We sought to examine the effects of modified venovenous ultrafiltration after cardiopulmonary bypass on pulmonary compliance in infants. METHODS: We prospectively enrolled 38 infants undergoing their first operation for congenital heart disease. Infants were randomized to receive 20 minutes of modified ultrafiltration after bypass or control. Static and dynamic compliance was measured after induction of anesthesia, before and immediately after filtration in the operating theater, 1 hour after return to the pediatric intensive care unit, and 24 hours after the operation. Length of time on the ventilator, inotropic requirements, and length of stay in the intensive care unit were recorded. RESULTS: Modified ultrafiltration produced a significant immediate improvement in dynamic (pre-ultrafiltration 2.5 +/- 1.9 mL/cm H(2)O to post ultrafiltration 2.9 +/- 2.7 mL/cm H(2)O, P =.03) and static (pre-ultrafiltration 2.1 +/- 0.9 mL/cm H(2)O to post-ultrafiltration 2.9 +/- 2.1 mL/cm H(2)O, P =.04) compliance. However, there was no significant difference in the change in dynamic (P =.3) or static (P =.7) compliance in the ultrafiltration and control groups when compared before the operation, after the operation, and at 24 hours. There was no significant difference in the time to extubation between patients and control subjects (140 +/- 91 hours vs 90 +/- 58 hours) or the length of intensive care unit stay (10.0 +/- 9.1 days vs 7.4 +/- 5.7 days). CONCLUSIONS: Modified ultrafiltration produces an improvement in pulmonary compliance after bypass in infants. However, these improvements are not sustained past the immediate post ultrafiltration period and do not lead to a decreased length of intubation or intensive care unit stay. PMID- 10694611 TI - Repair of the truncal valve and associated interrupted arch in neonates with truncus arteriosus. AB - OBJECTIVE: Truncal valve regurgitation and interrupted aortic arch have frequently been identified as risk factors in the repair of truncus arteriosus. We wished to examine these factors in the current era including the impact of truncal valve repair. METHODS: Between January 1992 and August 1998, 50 patients underwent surgical repair of truncus arteriosus. Their ages ranged from 2 days to 6 months (median, 2 weeks). Nine patients had associated interrupted aortic arch. Of the 14 patients (28%) in whom truncal valve regurgitation was diagnosed preoperatively, 5 had mild regurgitation, 5 had moderate regurgitation, and 4 had severe regurgitation. Five underwent truncal valve repair and 1 underwent homograft replacement of the truncal valve with coronary reimplantation. RESULTS: The actuarial survival was 96% at 30 days, 1 year, and 3 years. There were no deaths in patients with associated interrupted aortic arch. The 2 deaths in the series occurred in patients with truncal valve regurgitation, neither of whom underwent repair. Postoperative transthoracic echocardiography in patients who underwent valve repair showed minimal residual valvular regurgitation. None of the patients has required reoperation because of truncal valve problems or aortic arch stenosis at a median follow-up of 23 months (range, 1-60 months). Conduit replacement has been done in 17 patients (34%) after a mean duration of 2 years. The freedom from reoperation for those who had an aortic homograft was 4 years and for those who had a pulmonary homograft was 3 years. CONCLUSION: Despite the magnitude of the operation, excellent results can be achieved in complex forms of truncus arteriosus. In the current era interrupted aortic arch is no longer a risk factor for repair of truncus. Aggressive application of truncal valvuloplasty methods should neutralize the traditional risk factor of truncal valve regurgitation. PMID- 10694612 TI - Does the degree of cyanosis affect myocardial adenosine triphosphate levels and function in children undergoing surgical procedures for congenital heart disease? AB - OBJECTIVE: The outcome of children with cyanosis after cardiac surgical procedures is inferior to that of children who are acyanotic. Animal studies indicated detrimental effects of chronic hypoxia on myocardial metabolism and function. We studied whether the presence or the degree of cyanosis adversely affected myocardial adenosine triphosphate, ventricular function, and clinical outcome in children. METHODS: Forty-eight children who underwent repair of tetralogy of Fallot were divided according to their preoperative saturation: group I, 90% to 100% (n = 14 patients); group II, 80% to 89% (n = 16 patients); and group III, 65% to 79% (n = 18 patients). Adenosine triphosphate was measured from right ventricular biopsy specimens taken before ischemia, at 15 minutes of ischemia, at end-ischemia, and at 15 minutes of reperfusion. Ejection fraction was measured by echocardiography. RESULTS: Even before surgical ischemia, compared with groups I and II, group III had lower preoperative ejection fraction (59% +/- 2.9% vs 67% +/- 1.7% and 68% +/- 1.0%; P <.01) and lower preischemic adenosine triphosphate levels (15.1 +/- 2.1 vs 19.1 +/- 1.9 and 21.4 +/- 1.5 micromol/g dry weight; P <.01). After 15 minutes of ischemia, group III had lower adenosine triphosphate levels (11.2 +/- 1.8 vs 14.77 +/- 2.3 and 17. 6 +/- 3.1 micromol/g dry weight; P <.01). With reperfusion, both cyanotic groups lost further adenosine triphosphate compared with partial recovery in the acyanotic group (-22% +/- 3.8%, -20% +/- 3. 1% vs +18% +/- 1.8%; P <.01). Children in group III had a more complicated postoperative course as evidenced by longer ventilatory support (85 +/- 25 hours vs 31 +/- 15 and 40 +/- 21 hours; P =.07), inotropic support (86 +/- 23 hours vs 38 +/- 12 and 36 +/- 4 hours; P <.01), and intensive care unit stay (160 +/- 35 hours vs 60 +/- 10 and 82 +/- 18 hours; P =.02). CONCLUSIONS: The degree of cyanosis adversely affects myocardial adenosine triphosphate, function, and clinical outcome of children who undergo cardiac operation. Children with cyanosis should be identified as a higher risk group that could be targeted for supportive interventions. PMID- 10694613 TI - Oxygen consumption after cardiopulmonary bypass surgery in children: determinants and implications. AB - OBJECTIVE: We sought to assess oxygen consumption and its determinants in children shortly after undergoing cardiopulmonary bypass operations. METHODS: Twenty children, aged 2 months to 15 years (median, 3.75 years), undergoing hypothermic cardiopulmonary bypass operations were studied during the first 4 hours after arrival in the intensive care unit. Central and peripheral temperatures were monitored. Oxygen consumption was continuously measured by using respiratory mass spectrometry. Oxygen delivery was calculated from oxygen consumption and arterial and mixed venous oxygen contents, which were sampled every 30 minutes. Oxygen extraction was derived by the ratio of oxygen consumption and oxygen delivery. Arterial blood lactate levels were measured every 30 minutes. RESULTS: There was a correlation between oxygen consumption and age in patients older than 3 months (r = -0.76). Mean oxygen consumption increased by 14.7% during the study. The increase in oxygen consumption was correlated with the increase in central temperature (r = 0.73). Nine patients had an arterial lactate level above 2 mmol/L on arrival. There were no significant differences in oxygen consumption, oxygen delivery, and oxygen extraction between the group with lactate levels between 2 and 3 mmol/L and the groups with normal lactate levels both on arrival and at 2 hours. One patient with a peak lactate level of 6.8 mmol/L had initially low oxygen delivery (241.3 mL. min(-1). m(-2)). CONCLUSIONS: During the early hours after a pediatric cardiac operation, the increase in oxygen consumption is mainly attributed to the increase in central temperature. Oxygen consumption is negatively related to age. Mild lactatemia is common and does not appear to reflect oxygen delivery or oxygen consumption or a more complicated recovery. PMID- 10694614 TI - Vascular endothelial growth factor and basic fibroblast growth factor in children with cyanotic congenital heart disease. AB - OBJECTIVE: Vascular endothelial growth factor and basic fibroblast growth factor are potent stimulators of angiogenesis. Children with cyanotic congenital heart disease often experience the development of widespread formation of collateral blood vessels, which may represent a form of abnormal angiogenesis. We undertook the present study to determine whether children with cyanotic congenital heart disease have elevated serum levels of vascular endothelial growth factor and basic fibroblast growth factor. METHODS: Serum was obtained from 22 children with cyanotic congenital heart disease and 19 children with acyanotic heart disease during cardiac catheterization. Samples were taken from the superior vena cava, inferior vena cava, and a systemic artery. Vascular endothelial growth factor and basic fibroblast growth factor levels were measured in the serum from each of these sites by enzyme-linked immunosorbent assay. RESULTS: Vascular endothelial growth factor was significantly elevated in the superior vena cava (P =.04) and systemic artery (P =.02) but not in the inferior vena cava (P =.2) of children with cyanotic congenital heart disease compared to children with acyanotic heart disease. The mean vascular endothelial growth factor level, determined by averaging the means of all 3 sites, was also significantly elevated (P =.03). Basic fibroblast growth factor was only significantly elevated in the systemic artery (P =.02). CONCLUSION: Children with cyanotic congenital heart disease have elevated systemic levels of vascular endothelial growth factor. These findings suggest that the widespread formation of collateral vessels in these children may be mediated by vascular endothelial growth factor. PMID- 10694615 TI - Transmyocardial laser revascularization combined with coronary artery bypass grafting: a multicenter, blinded, prospective, randomized, controlled trial. AB - OBJECTIVE: We sought to assess the safety and efficacy of transmyocardial revascularization combined with coronary artery bypass grafting in patients not amenable to complete revascularization by coronary bypass alone. METHODS: A total of 263 patients whose standard of care was coronary artery bypass grafting and who had one or more ischemic areas not amenable to bypass grafting were prospectively randomized to receive coronary bypass of suitable vessels plus transmyocardial revascularization to areas not graftable (n = 132) or coronary bypass alone with nongraftable areas left unrevascularized (n = 131). Group preoperative demographics and operative characteristics were similar. RESULTS: The operative mortality rate after coronary bypass/transmyocardial revascularization was 1.5% (2/132) versus 7.6% (10/131) after coronary bypass alone (P =.02). Patients undergoing both coronary bypass and transmyocardial revascularization required less postoperative inotropic support (30% vs 55%, P =.0001) and had a trend toward fewer insertions of intra-aortic balloon pumps (4% vs 8%, P =.13) than did patients having coronary bypass alone. Multivariable predictors of operative mortality were coronary artery bypass alone (odds ratio, 5.3; 95% confidence interval, 1.1-25.7; P =.04) and increased age (odds ratio, 1.1; 95% confidence interval, 1. 0-1.2; P =.03). One-year Kaplan-Meier survival (95% vs 89%, P =.05) and freedom from major adverse cardiac events defined as death or myocardial infarction (92% vs 86%, P =.09) favored the combination of coronary bypass and transmyocardial revascularization. Baseline to 12-month improvement in angina and exercise treadmill scores was similar between groups. CONCLUSIONS: In a prospective, randomized, multicenter trial, transmyocardial revascularization combined with coronary artery bypass grafting in patients not amenable to complete revascularization by coronary bypass alone was safe; however, angina relief and exercise treadmill improvement were indistinguishable between groups at 12 months of follow-up. Operative and 1-year survival benefits observed after adjunctive transmyocardial revascularization require confirmation by a larger validation study, which is ongoing. PMID- 10694616 TI - Results of revascularization in patients with severe left ventricular dysfunction. AB - OBJECTIVE: In patients with coronary disease and poor left ventricular function, bypass grafting remains a surgical challenge. This study evaluates experience in 125 consecutive patients with ejection fraction less than 20% (study group). METHODS: Preoperative viability studies were not used for patient selection. Clinical data were prospectively collected. The average age of the study subjects was 59 +/- 9 years, and 112 (90%) were male. Most patients (108 [86%]) were in symptom class III or IV. Main indications for surgery included angina in 62 (50%), heart failure and angina in 36 (29%), heart failure in 9 (7%), ventricular arrhythmia in 2 (2%), and critical anatomy in 16 (13%). Significant mitral regurgitation was present in 48 (38%), and distal vessels were poorly visualized in 67 (54%). At surgery, temperature mapping guided an integrated approach to cold cardioplegia. Results in this group were compared with those obtained in case-matched control subjects receiving cardioplegia without temperature mapping (matched for age, sex, functional class, and urgency of operation). RESULTS: Hospital morbidity (intra-aortic balloon pump support) and mortality rates were significantly lower in the study group versus those of control subjects (15% vs 30%, P =. 004; and 4% vs 11%, P =.03, respectively). In study patients the 5-year actuarial survival was 72%. Among survivors, both anginal class and heart failure class improved significantly. By means of multivariate analysis, survival was adversely affected by older age, class IV symptoms, and poorly visualized distal vessels. CONCLUSIONS: These results support the use of coronary artery bypass grafting in patients with severe left ventricular dysfunction without case selection on the basis of viability studies or visibility of distal vessels. Low hospital morbidity and mortality rates have been achieved when temperature mapping guides cardioplegia. Symptoms are improved in most patients, and long term survival is encouraging. PMID- 10694617 TI - Extended total arch replacement for acute type a aortic dissection: experience with seventy patients. AB - OBJECTIVE: We sought to report the clinical experience with extended total arch replacement for acute type A aortic dissection and to determine the factors that influence early mortality, late survival, and late reoperation. METHODS: Between December 1988 and August 1998, 70 patients underwent emergency graft replacement of both the ascending aorta and the total aortic arch for acute type A aortic dissection. All operations were performed with hypothermic extracorporeal circulation, selective cerebral perfusion for cerebral protection during aortic arch repair, and open distal anastomosis. Concomitant procedures included aortic valve resuspension in 18 patients, composite graft replacement in 10 patients, and coronary artery bypass grafting in 5 patients. RESULTS: The early mortality rate was 16% (11 of 70 patients). Multivariable analysis showed that renal mesenteric ischemia and coronary artery bypass grafting were independent determinants for early death. Survival rates at 3 and 5 years postoperatively, including the early deaths, were 75% +/- 5% and 73% +/- 6%, respectively. Multivariable analysis showed that renal-mesenteric ischemia and en bloc repair were independent determinants for late death. Freedom from reoperation was 91% +/ 4% and 77% +/- 8% at 3 and 5 years, respectively. Multivariable analysis showed that anastomotic leakage was the only significant determinant for late reoperation. CONCLUSIONS: Extended total arch replacement for acute type A aortic dissection could be justified in properly selected patients. PMID- 10694619 TI - Topical use of tranexamic acid in coronary artery bypass operations: a double blind, prospective, randomized, placebo-controlled study. AB - OBJECTIVES: We sought to investigate the effect of topical application of tranexamic acid into the pericardial cavity in reducing postoperative blood loss in coronary artery surgery. METHODS: A prospective, randomized, double-blind investigation with parallel groups was performed. Forty consecutive patients undergoing primary coronary surgery were randomly assigned to group 1 (tranexamic acid group) or group 2 (placebo group). Tranexamic acid (1 g in 100 mL of saline solution) or placebo was poured into the pericardial cavity and over the mediastinal tissues before sternal closure. The drainage of mediastinal blood was measured hourly. RESULTS: Chest tube drainage in the first 24 hours was 485 +/- 166 mL in the tranexamic acid group and 641 +/- 184 mL in the placebo group (P =.01). Total postoperative blood loss was 573 +/- 164 mL and 739 +/- 228 mL, respectively (P =.01). The use of banked donor blood products was not significantly different between the two groups. Tranexamic acid could not be detected in any of the blood samples blindly collected from 24 patients to verify whether any systemic absorption of the drug occurred. There were no deaths in either group. None of the patients required reoperation for bleeding. CONCLUSIONS: Topical application of tranexamic acid into the pericardial cavity after cardiopulmonary bypass in patients undergoing primary coronary bypass operations significantly reduces postoperative bleeding. Further studies must be carried out to clarify whether a more pronounced effect on both bleeding and blood products requirement might be seen in procedures with a higher risk of bleeding. PMID- 10694618 TI - Sodium nitroprusside in patients with compromised left ventricular function undergoing coronary bypass: reduction of cardiac proinflammatory substances. AB - OBJECTIVE: The aim of the present study was to investigate whether the nitric oxide donor sodium nitroprusside can reduce the cardiac inflammatory response during coronary artery bypass grafting in patients with severely compromised left ventricular function. METHODS: Patients (n = 30) were assigned to receive placebo or sodium nitroprusside (0.5 microg. kg(-1). min(-1)) for the first 60 minutes of reperfusion. Interleukin 6, interleukin 8, and tumor necrosis factor alpha levels; platelet adhesion molecule CD41 and CD62 levels; and CD11b on leukocytes were determined in the radial artery and coronary sinus before cardiopulmonary bypass and during reperfusion (1, 5, 10, 35, and 75 minutes). RESULTS: At 1 minute of reperfusion, coronary venous levels of CD41-positive polymorphonuclear leukocytes were 8% lower than arterial levels in the placebo group and 18% higher in the sodium nitroprusside group (P =.021). At 5 minutes of reperfusion, the respective levels were 29% and 1% for interleukin 6 (P =.015), -5% and 20% for CD41-positive monocytes (P =.032), and -2% and 16% for CD11b-positive monocytes (P =.038). At 10 minutes of reperfusion, these levels were -14% and 21% for CD41 positive monocytes (P =.006). At 35 minutes of reperfusion, these levels were 13% and 7% for CD41-positive monocytes (P =.017), -41% and 23% for CD11b-positive monocytes (P =.001), and 7% and 25% for CD62-positive platelets (P =. 041). At 75 minutes of reperfusion, the levels were 15% and -7% for tumor necrosis factor alpha (P =.025) and -10% and 10% for CD62-positive platelets (P =.041). CONCLUSIONS: Transcardiac production of proinflammatory cytokines is reduced in patients undergoing coronary artery bypass grafting treated with the nitric oxide donor sodium nitroprusside. At the same time, less activated leukocytes and platelets are retained in the coronary circulation. PMID- 10694620 TI - Novacor left ventricular assist system versus Heartmate vented electric left ventricular assist system as a long-term mechanical circulatory support device in bridging patients: a prospective study. AB - OBJECTIVE: Long-term mechanical circulatory support as a bridge-to transplantation procedure and bridge to recovery is of increasing importance. The implantable left ventricular assist devices, Novacor N100 left ventricular assist system (Baxter Healthcare Corporation, Berkeley, Calif) and TCI HeartMate vented electric left ventricular assist system (Thermo Cardiosystems Inc, Woburn, Mass), have proved to be efficient devices in bridge-to-transplantation settings and for prolonged support. The two systems were compared with regard to reliability and morbidity. METHODS: Between October 1996 and March 1998, a prospective, single center study was done that included 40 patients, 20 of whom were treated with the Novacor system and 20 of whom were treated with the HeartMate device. The diseases were mainly dilated cardiomyopathy (13/9) and ischemic cardiomyopathy (6/10). There were no statistically significant differences between the two groups regarding age, sex, preoperative clinical blood chemistry values, hemodynamic data, or risk factors. RESULTS: There were no statistically significant differences between the two groups with regard to postoperative hemodynamics, organ recovery, out-of-hospital support, and survival to heart transplantation. Mean duration of support was 235.3 +/- 210 days for the Novacor group and 174.6 +/- 175 days for the HeartMate group and mean out-of-hospital support was 241 +/- 179 days and 166 +/- 152 days for the two groups, respectively. Neurologic complications occurred significantly more often among the Novacor group, whereas the HeartMate group had a higher prevalence of infections and technical problems, which was statistically significant. Survival to transplantation was 65% for the Novacor group and 60% for the HeartMate group. CONCLUSIONS: Most patients had organ recovery with left ventricular assist system support, and a considerable number of patients in both groups underwent transplantation. However, both devices need revision to address the current problems, that is, thromboembolism for the Novacor device and infection and reliability for the HeartMate device. PMID- 10694621 TI - Hypothermic retrograde venous perfusion with adenosine cools the spinal cord and reduces the risk of paraplegia after thoracic aortic clamping. AB - OBJECTIVE: We evaluated the utility of retrograde venous perfusion to cool the spinal cord and protect neurologic function during aortic clamping. We hypothesized that hypothermic adenosine would preserve the spinal cord during ischemia. METHODS: Six swine (group I) underwent thoracic aortic occlusion for 30 minutes at normothermia. Group II animals underwent spinal cooling by retrograde perfusion of the paravertebral veins with hypothermic (4 degrees C) saline solution during aortic occlusion. The spinal cords of group III animals were cooled with a hypothermic adenosine solution in a similar fashion. Intrathecal temperature was monitored and somatosensory evoked potentials assessed the functional status of spinal pathways. RESULTS: Spinal cooling without systemic hypothermia significantly improved neurologic Tarlov scores in group III (4.8 +/- 0.2) and group II (3.8 +/- 0.4) when compared with group I scores (1.3 +/- 0.6) (P <.001). Furthermore, 5 of the 6 animals in group III displayed completely normal neurologic function, whereas only one animal in group II and no animals in group I did (P =.005). Somatosensory evoked potentials were lost 10.6 +/- 1.4 minutes after ischemia in group I. In contrast, spinal cooling caused rapid cessation of neural transmission with loss of somatosensory evoked potentials at 6.9 +/- 1.2 minutes in group II and 7.0 +/- 0.8 minutes in group III (P =.06). Somatosensory evoked potential amplitudes returned to 85% of baseline in group III and 90% of baseline in group II compared with only 10% of baseline in group I (P =.01). CONCLUSIONS: We conclude that retrograde cooling of the spinal cord is possible and protects against ischemic injury and that adenosine enhances this effect. The efficacy of this method may be at least partly attributed to a more rapid reduction in metabolic and electrical activity of the spinal cord during ischemia. PMID- 10694622 TI - Single-center experience with the thoratec ventricular assist device. AB - OBJECTIVE: The Thoratec ventricular assist device (Thoratec Laboratories, Pleasanton, Calif) is widely accepted for univentricular and biventricular support in patients with various indications. The aim of this study is to describe our experience with implantation of the Thoratec ventricular assist device in more than 100 patients. METHODS: From March 1992 to June 1998, 114 patients (98 men and 16 women; mean age, 47.9 years) received the Thoratec ventricular assist device for a mean duration of 44.9 days. The patients were divided into 3 groups. Group 1 included 84 patients in whom the system was applied as a bridge-to-transplant procedure. Group 2 included 17 patients with postcardiotomy cardiogenic shock, and group 3 included 13 patients with cardiogenic shock of other causes. RESULTS: Sixty-eight percent of patients in group 1 survived to transplantation with a posttransplant survival of 88%. The only independent risk factor affecting survival was age more than 60 years. Survivals in groups 2 and 3 were 47% and 31%, respectively. Main complications in all groups were bleeding, multiple organ failure, liver failure, sepsis, and neurologic disorders. CONCLUSIONS: The Thoratec ventricular assist device has proved to be a reliable device for bridge to transplantation and postcardiotomy support. Further studies are required on patient selection and on patient and device management to reduce the incidence of complications in these patient populations. PMID- 10694623 TI - Mechanisms of myocardial protection by adenosine-supplemented cardioplegic solution: myofilament and metabolic responses. AB - OBJECTIVE: Adenosine supplementation of cardioplegic solutions in cardiac operations improves postarrest myocardial recovery after cardioplegic arrest and reperfusion; however, the mechanism of the action of adenosine remains unknown. We tested the hypotheses that adenosine-supplemented cardioplegic solution improves myofibrillar protein cooperative interaction and increases myocardial anaerobic glycolysis. METHODS: The hearts of male Sprague-Dawley rats were randomized to undergo 120 minutes of cardioplegic arrest with 1 of 3 cardioplegic solutions: (1) St Thomas' Hospital No. 2 cardioplegic solution (St Thomas group), (2) St Thomas' Hospital No. 2 cardioplegic solution plus adenosine (100 micromol/L) (adenosine group), and (3) St Thomas' Hospital No. 2 cardioplegic solution plus adenosine (100 micromol/L) plus the nonspecific adenosine receptor antagonist 8-p -sulfophenyltheophylline (50 micromol/L) (sulfophenyltheophylline group). A fourth group of hearts underwent no cardioplegic arrest. RESULTS: Systolic and diastolic functional recovery was improved in the adenosine group compared with that in the other two groups, independent of coronary flow. Adenosine supplementation of cardioplegic solution prevented the decrease in myofibrillar protein cooperative interaction seen after cardioplegic arrest and reperfusion (St Thomas and sulfophenyltheophylline groups). Adenosine supplemented cardioplegic solution also caused significantly increased anaerobic glycolysis during cardioplegic arrest. These responses were blocked in the sulfophenyltheophylline group. CONCLUSIONS: The changes in myocardial glycolytic activity and myofilament cooperativity coincided with functional recovery in the three cardioplegia groups and may represent mechanisms underlying protection with adenosine-supplemented cardioplegic solution. PMID- 10694624 TI - Intermediate lukewarm (20 degrees c) antegrade intermittent blood cardioplegia compared with cold and warm blood cardioplegia. AB - BACKGROUND: In the field of intermittent antegrade blood cardioplegia, 3 levels of temperature are commonly used: (1) cold (8 degrees C); (2) tepid (29 degrees C); and (3) warm (37 degrees C). Given the 21 degrees C spread and the metabolic changes that can occur between cold (8 degrees C) and tepid (29 degrees C) cardioplegia, we thought it worthwhile to test a temperature halfway between the cold and tepid levels. The aim of this study was to test the quality of myocardial protection provided by intermediate lukewarm (20 degrees C) cardioplegia by comparing it with cold and warm cardioplegia. Protection was assessed by measuring cardiac troponin I release. METHODS: One hundred thirty five patients undergoing coronary artery bypass grafting were enrolled in a prospective randomized trial comparing cold (8 degrees C), intermediate lukewarm (20 degrees C), and warm (37 degrees C) antegrade intermittent blood cardioplegia. Cardiac troponin I concentrations were measured in serial venous blood samples. RESULTS: The total amount of cardiac troponin I released was significantly higher in the cold group (4.7 +/- 2.3 microg) than in the intermediate lukewarm (3.4 +/- 2.0 microg) or the warm (3.1 +/- 2.7 microg) groups. The cardiac troponin I concentration was significantly higher at hour 6 in the intermediate lukewarm group (1. 23 +/- 0.55 microg/L) than in the warm group (0.89 +/- 0.50 microg/L). CONCLUSIONS: Intermittent antegrade intermediate lukewarm blood cardioplegia is appropriate and clinically safe. Cardiac troponin I release suggests that intermediate lukewarm cardioplegia is better than cold cardioplegia but less effective than warm cardioplegia in low-risk patients. We therefore recommend the use of warm cardioplegia in low-risk patients. PMID- 10694625 TI - Brain function monitoring during bidirectional Glenn procedures. PMID- 10694626 TI - Endovascular stent-graft repair with a flexible stent-graft and a simple applicator prepared from a syringe. PMID- 10694627 TI - Lymphangioleiomyomatosis: the surgeon's role in diagnosis. PMID- 10694628 TI - Clinical-pathologic conference in thoracic surgery: Masaoka stage 2-a thymoma. PMID- 10694629 TI - Venovenous modified ultrafiltration after cardiopulmonary bypass in children: A prospective randomized study PMID- 10694630 TI - Alternate explanation of the additive protective effects of regional infusion of hypothermic saline and adenosine solutions. PMID- 10694631 TI - Should methylene blue be the drug of choice to treat vasoplegias caused by cardiopulmonary bypass and anaphylactic shock? PMID- 10694632 TI - Should methylene blue be the drug of choice to treat vasoplegias caused by cardiopulmonary bypass and anaphylactic shock? PMID- 10694633 TI - Combined mitral valve repair and the cox maze procedure for mitral valve prolapse and regurgitation and associated atrial fibrillation. PMID- 10694634 TI - Should the bidirectional glenn procedure be better performed through the support of cardiopulmonary bypass? PMID- 10694635 TI - Should the bidirectional glenn procedure be better performed through the support of cardiopulmonary bypass? PMID- 10694636 TI - Preimplantation retrograde pneumoplegia in clinical lung transplantation. PMID- 10694637 TI - Preimplantation retrograde pneumoplegia in clinical lung transplantation PMID- 10694638 TI - Retirement of journal editor PMID- 10694639 TI - Children will benefit as pediatric groups merge: a surgical oncologist's perspective. PMID- 10694640 TI - Do the newer imaging modalities affect management of solid tumors? PMID- 10694641 TI - Prognostic significance of mucinous carcinoma of colon and rectum: a prospective case-control study. AB - BACKGROUND AND OBJECTIVES: The clinical meaning of mucinous type of colonic and rectal carcinoma is still controversial. We used clinicopathological and follow up data prospectively recorded for our series of colon and rectum cancer to compare 2 matched groups of mucinous and nonmucinous cancer patients. METHODS: Two-hundred-forty-eight patients operated for colon and rectum cancer between January 1986 and January 1997 were considered. Thirty-six patients showed mucinous pattern on histologic examination but only 29 (11.7%) had more than 50% of mucin-secreting acini and could be classified as mucinous type. The 29 mucinous cancer patients were compared with 212 nonmucinous cancer patients to evaluate differences in epidemiological and clinical features. A control group from the nonmucinous patients was sorted by matching for age, sex, location, and Dukes stage. RESULTS: In the case-control groups, survival was better for nonmucinous than for mucinous tumours. Many of the epidemiological findings already observed for mucinous carcinoma were also confirmed. CONCLUSIONS: The existence of prognostic, clinical, and epidemiological differences between mucinous and nonmucinous colorectal carcinoma, together with the preliminary reports about their difference as to genetic features, could support the hypothesis that mucinous type is a distinct biological entity. PMID- 10694642 TI - Internal mammary chain sentinel lymph node identification in breast cancer. AB - BACKGROUND AND OBJECTIVES: Sentinel lymph node (SLN) biopsy is not usually performed with respect to the internal mammary lymph node chain. However, the SLN may be located in the internal mammary chain, particularly with medial lesions. We carried out this study to investigate whether lymphatic mapping and SLN biopsy can detect internal mammary involvement in patients with breast cancer. METHODS: A dye- and gamma probe-guided SLN biopsy was performed in a consecutive series of 41 patients with tumor in situ or clinical stage I or II breast cancer. After the biopsy, these patients underwent either a modified radical mastectomy or breast conserving surgery including axillary lymph node dissection. Biopsy of internal mammary lymph nodes was performed in 19 of these patients. RESULTS: No involvement of internal mammary lymph nodes was found histologically in 5 patients in whom lymphatic flow or a "hot nodule" in the internal mammary chain was found using lymphoscintigraphy. Nodal involvement was demonstrated histologically in only 1 of 5 cases where lymphatic vessels showed dye staining or faintly stained nodes. Internal mammary lymph node biopsy also was performed in 14 of 36 patients with neither stained lymphatic vessels or nodes, nor with lymphatic flow or a hot nodule by lymphoscintigraphy. Nodal involvement was found histologically in 1 of these patients. CONCLUSION: SLN biopsy guided by lymphatic mapping is unreliable for identifying metastases to internal mammary lymph nodes. PMID- 10694643 TI - Clinical importance of late recurrence in soft-tissue sarcomas. AB - BACKGROUND AND OBJECTIVES: Soft-tissue sarcomas (STS) represent a diverse histologic group of malignancies at risk for local and distant failure. We studied the impact of late (5 or more years) vs. early recurrence (less than 5 years) on subsequent outcome. METHODS: Four hundred sixty-eight patients with STS treated between 1962 and 1992 were evaluated for late (n = 39; 8%) or early (n = 253; 54%) recurrence. Clinical and pathologic factors were reviewed. Survival data were analyzed by the Kaplan-Meier method and the log-rank test. RESULTS: Of the 39 patients with a late recurrence (median follow-up 156 months), 18 patients had local recurrence, 7 patients developed distant recurrence, and 14 patients had local and distant recurrence. Thirty patients with late local and/or distant recurrence underwent complete or wide excision (n = 16), amputation (n = 4), or local resection (n = 10). The overall 5-year survival rate following late recurrence was 61%. The 5-year overall survival rate was statistically better for patients with a late local recurrence alone than for patients with distant failure, 94% vs. 36%, respectively (P = 0.003). Neither the site of the primary STS, age, primary margin status, nor histology had any effect on subsequent local or distant failure and subsequent survival. CONCLUSIONS: These data suggest that an aggressive approach is appropriate in patients who present with late recurrence (more than 5 years) following treatment of the primary STS. Impressive survival rates can be achieved in the treatment of local recurrences. PMID- 10694644 TI - Limb-saving surgery, survival, and prognostic factors for osteosarcoma: the Hungarian experience. AB - BACKGROUND AND OBJECTIVES: There are many factors thought to have an influence on the prognosis of osteosarcoma that have been reported in the literature. Their significance, however, still remains controversial in most cases. Experience with osteogenic sarcoma (OS) was reviewed in order to evaluate surgical results and survival and to determine the prognostic factors. METHODS: Ninety-six patients with high-grade osteosarcoma of the extremities were treated between 1986 and 1997 in the authors' institution. They were divided into 3 groups: In group I, all 75 patients with nonmetastatic OS received intensive chemotherapy (high-dose methotrexate, doxorubicin, ifosfamide, and cisplatin) and underwent surgery. In group II, 9 patients already had metastases at the time of referral. In group III, 12 patients received chemotherapy in delayed or suboptimal form. Results and Conclusions In group I, there were local recurrences in 3 patients (7%) and metastases in 8 patients (20%) with limb-saving, whereas these numbers were 1 (3%) and 14 (38%) in those who had amputation. The 5-year disease-free survival (DFS) was 72% and 69% in the limb-saving and amputation groups, respectively. In groups II and III, 5-year DFS was extremely poor, 10% and 20% only, underlining the importance of stage and intensity of chemotherapy, respectively. With univariate analysis, sex, duration of symptoms, and radiographic appearance of OS had no prognostic value, whereas tumor volume <60 cm(3), wide or radical surgical margin, distal location of OS, cartilagineous ground substance <20%, and response to chemotherapy were positive prognostic factors. The last 4 variables maintained their significance in the multivariate Cox model as well. Age >30 years showed indirect negative influence on the final outcome through enhanced intolerability to the drugs and less cooperability of the patients. The results on survival with limb-saving surgery were well comparable with those of amputation. PMID- 10694645 TI - Expression of matrix metalloproteinases and the tissue inhibitors of metalloproteinases and their local invasiveness and metastasis in Chinese human pancreatic cancer. AB - BACKGROUND AND OBJECTIVES: The objective was to evaluate the potent role of matrix metalloproteinases(MMPs) and the tissue inhibitors of metalloproteinases(TIMPs) in processes leading to metastasis and local invasiveness of Chinese human ductal adenocarcinomas of the pancreas. We also evaluated a possible biological association between the gene expression and clinical manifestations. METHODS: Northern blot and in situ hybridization have shown MMP and TIMP gene expression in the pancreas and alterations associated with neoplastic transformation. Fifteen cases of surgical pancreatic specimens were examined, using cDNA probes to MMP2, MMP9, and TIMP1. Findings were correlated with the size of tumor section, CA19-9, pathological classification, thrombosis, and infiltration of capsule and lymphonoids. RESULTS: Increased levels of the mRNA of MMP2, MMP9, and TIMP1 genes, MMP2 approximately MMP9/= 0.70. RESULTS: Six patients (50%) demonstrated MIN for at least 1 locus, and 2 patients demonstrated loss of heterozygosity (LOH) for at least 1 locus. Only 1 individual's chordoma manifested microsatellite instability (MIN) and loss of heterozygosity (LOH). Another patient manifested no MIN but LOH at 9p and 18q. Interestingly, this individual had the most aggressive clinical cancer course, presenting with lymph node metastasis and succumbing to widespread metastatic disease. CONCLUSIONS: Chordomas can be added to the list of malignancies demonstrating MIN. LOH may prove to portend a worse prognosis than MIN when more tumors are examined. PMID- 10694647 TI - Clinical significance of retinoblastoma protein (pRB) expression in esophageal squamous cell carcinoma. AB - BACKGROUND AND OBJECTIVES: The goal was to evaluate the clinicopathological significance of retinoblastoma gene product (pRB) expression in esophageal squamous cell carcinoma. METHODS: We investigated abnormal pRB expression in tumors in 191 patients using an immunohistochemical method in conjunction with anti-RB protein antibody. Surgically resected esophageal squamous cell carcinomas were examined by immunohistochemical analysis for altered pRB expression. RESULTS: Decreased pRB nuclear staining indicating loss of RB function occurred in 82 (43%) of the cases studied. The incidence of decreased pRB expression was higher in tumors with invasion to the adventitia (50%) than in tumors without invasion to the adventitia (33%, P = 0.0188). In addition, the incidence of decreased pRB expression was higher in tumors with lymph node metastasis (50%) than in those without (34%, P = 0.0346). The 3-year survival rates of 82 patients who had tumors with decreased pRB expression (30%) was significantly lower than that of 109 patients who had tumors with normal pRB expression (52%, P = 0.0032). However, in the multivariate survival analysis, pRB expression was not an independent prognostic factor for patients with esophageal squamous cell carcinoma. CONCLUSIONS: Abnormal pRB expression appears to be closely associated with tumor development. However, the existence of tumors with hyperphosphorylated RB protein (inactivated form) in pRB-positive tumors, such as those in the present study, should be considered. Thus, discrimination of this hyperphosphorylated form of RB protein from the unphosphorylated RB protein is needed. PMID- 10694648 TI - Transarterial chemoembolization for inoperable hepatocellular carcinoma and postresection intrahepatic recurrence. AB - BACKGROUND AND OBJECTIVES: The role of transarterial chemoembolization (TACE) for inoperable hepatocellular carcinoma (HCC) has remained controversial, and its efficacy for postresection intrahepatic recurrence has not been fully assessed. A study was performed to evaluate the treatment results and prognostic factors of TACE treatment in these patients. METHODS: Clinicopathologic data and treatment results of 384 patients with inoperable HCC and 100 patients with postresection recurrent HCC treated with TACE were collected prospectively and analyzed. RESULTS: TACE was associated with an overall treatment morbidity rate of 23% (112/484) and mortality rate of 4.3% (21/484). A particularly high mortality rate of 20% (9/45) was observed among patients with tumors > 10 cm and pretreatment serum albumin level 35 g/L were independent favorable prognostic factors. TACE in patients with postresection recurrent HCC was associated with less morbidity, mortality, and a better survival outcome compared with patients with primary inoperable HCC, but this was largely related to smaller tumor size and better liver function in the former group at the time of TACE treatment. CONCLUSIONS: TACE in patients with inoperable HCC was associated with significant morbidity and mortality, and the survival benefit was limited. Better patient selection in terms of tumor size and liver function may improve treatment results. Patients who have a tumor > 10 cm and poor liver function (serum albumin 1 in 250) at 15-18 weeks of gestation (study group) were compared with data from 272 age and gestation matched controls with a negative Down syndrome screening test from the same population. The frequency of normal and adverse perinatal outcomes, including pre-eclampsia, isolated intrauterine growth restriction, spontaneous preterm labour and stillbirth was recorded. The incidence of adverse pregnancy outcomes was 11.9% in the study group and 8.6% in the control group. The estimated odds ratio of an abnormal outcome in the study group was 1.41 (95% CI-0.790, 2.55). The observed difference between proportion was 0.0324 (95% CI-0.022, 0.083; p=0.40). These data identify no evidence for a strong association between a false positive Down syndrome screening test result and subsequent adverse perinatal outcomes in the general population. PMID- 10694682 TI - Maternal serum second trimester AFP and hCG in pregnancies with placenta previa. AB - This study was undertaken to investigate the association between placenta previa and Down syndrome screening analytes-alpha-fetoprotein (AFP) and hCG-during the second trimester. Measurements of maternal serum AFP and hCG concentrations were retrospectively analysed in relation to placenta previa in a cohort of 46 consecutive singleton pregnancies affected by placenta previa from January 1993 through December 1997 at the University Hospital of Kuopio, Finland, and then compared with those in healthy singleton control pregnancies (N=9560) from the same clinic over the same period of time. Geometric means of maternal serum AFP and hCG concentrations in pregnancies with placenta previa were 1.13 and 0. 85 multiples of the median (MoM), respectively. The mean maternal age was higher in the subjects than in the controls (30.9 years compared with 28.8 years) (p<0.01). In relation to Down syndrome risk assessment, the pattern of the two markers together with maternal age indicated high risk as often in the study subjects as in the controls. Even though the maternal serum AFP and hCG differences were not statistically significant, they may be of some clinical importance, and further studies are needed to evaluate whether placental site should be taken into account in maternal serum screening. PMID- 10694683 TI - Common trisomy mosaicism diagnosed in amniocytes involving chromosomes 13, 18, 20 and 21: karyotype-phenotype correlations. AB - Karyotype-phenotype correlations of common trisomy mosaicism prenatally diagnosed via amniocentesis was reviewed in 305 new cases from a collaboration of North American cytogenetic laboratories. Abnormal outcome was noted in 10/25 (40%) cases of 47,+13/46, 17/31 (54%) cases of 47,+18/46, 10/152 (6.5%) cases of 47,+20/46, and in 49/97 (50%) cases of 47,+21/46 mosaicism. Risk of abnormal outcome in pregnancies with less than 50% trisomic cells and greater than 50% trisomic cells were: 26% (4/15) versus 60% (6/10) for 47,+13/46, 52% (11/21) versus 75% (6/8) for 47,+18/46, 4.5% (6/132) versus 20% (4/20) 47,+20/46, and 45% (27/60) versus 59% (22/37) for 47,+21/46. Phenotypically normal liveborns were observed with mean trisomic cell lines of 9.3% for 47,+13/46, 8.6% for 47,+18/46, 27% for 47, +20/46, and 17% for 47,+21/46. Cytogenetic confirmation rates were 46% (6/13 cases) for 47,+13/46 mosaicism, 66% (8/12 cases) for 47, +18/46, 10% (10/97 cases) for 47,+20/46, and 44% (24/54 cases) for 47,+21/46. There were higher confirmation rates in pregnancies with abnormal versus normal outcome: 50% versus 44% for 47,+13/46 mosaicism, 100% versus 33% for 47,+18/46, 66% versus 7% for 47, +20/46, and 55% versus 40% for 47,+21/46. Repeat amniocentesis is not helpful in predicting clinical outcome. It may be considered when there is insufficient number of cells or cultures to establish a diagnosis. Fetal blood sampling may have a role in mosaic trisomy 13, 18, and 21 as the risk for abnormal outcome increases with positive confirmation: 1/5 (20%) normal cases versus 5/8 (62%) abnormal cases. High resolution ultrasound examination(s) is recommended for clinical correlation and to facilitate genetic counselling. PMID- 10694684 TI - Analysis of uncultured amniocytes by comparative genomic hybridization: a prospective prenatal study. AB - Comparative genomic hybridization (CGH) is a new molecular cytogenetic technique which can detect and map whole and partial aneuploidies throughout a genomic specimen DNA without culturing specimen cells. Thus, CGH may be used as a comprehensive and rapid screening test in prenatal unbalanced chromosomal abnormalities detection. We report the results of the first prospective study to evaluate the use of the CGH technique on uncultured amniocytes. Seventy-one amniotic fluid samples, obtained by transabdominal amniocentesis between the 14th and 35th weeks of gestation, were simultaneously investigated using CGH and conventional cytogenetics. Amniocentesis were done for advanced maternal age (21.1%), fetal ultrasound anomalies (73.3%) and high level of biochemical markers in maternal serum (5.6%). Sixty-six (93%) informative results were generated on a total of 71 analysed specimens. Fifty-nine samples were reported as disomic for all autosomes with a normal sex chromosome constitution using CGH and conventional cytogenetics. Among them, three pericentromeric chromosomal inversions were undetected by CGH analysis. Seven numerical aberrations were characterized, including one case of trisomy 13, one case of trisomy 18 and five cases of trisomy 21. Advantages and limitations of CGH for a rapid prenatal screening of unbalanced chromosomal aberrations are discussed. PMID- 10694685 TI - Haplotype association and mutation analysis of the transglutaminase 1 gene for prenatal exclusion of lamellar ichthyosis. AB - Lamellar ichthyosis (LI) is an autosomal recessive keratinization disorder of the skin. Genetic heterogeneity has been shown for the disease and there is evidence for the involvement of the transglutaminase 1 (TGM1) gene on chromosome 14q11. We have previously identified chromosome 14q11 haplotypes associated with ichthyosis in the Norwegian population. In this paper we describe antenatal exclusion of ichthyosis in two Norwegian families by chromosome 14q11 haplotype association and direct mutation analysis. In one pregnancy, the 11-week old fetus at risk for LI was found to share only one disease-associated haplotype. A subsequent mutation analysis of the TGM1 gene in fetal DNA revealed that the fetus carried a novel 3795A-->T transversion. The affected proband was compound heterozygous for the mutations 3795A-->T and 3239G-->C resulting in an Asp430Val and a Val379Leu, respectively. In another LI family, the 11-week old fetus was found to be heterozygous for the 14q11 haplotype associated with the disease. Subsequent mutation analysis revealed that the fetus was heterozygous for the 2526A-->G transition in the splice site of intron 5 whereas the proband was homozygous for the same mutation. Our results show that haplotyping can be a useful tool for prenatal diagnosis in diseases with genetic heterogeneity. PMID- 10694686 TI - Characterization and clinical implications of marker chromosomes identified at prenatal diagnosis. AB - Eighteen fetuses with marker chromosomes were detected at diagnostic amniocentesis in our laboratory among 15 781 amniocentesis samples. Using combined approaches, conventional cytogenetics including special stain techniques and fluorescence in situ hybridization (FISH), we successfully characterized 15 of them, which assisted subsequent genetic counselling. Six marker chromosomes were of sex chromosome origin, each of which substituted a missing sex chromosome, and 12 were supernumerary marker chromosomes (SMCs). Nine of the SMCs were proven to be of autosomal origin. Of those autosomal SMCs, five originated from chromosome 15, two from chromosome 18, one from chromosome 12 and one from chromosome 1. Among 16 marker chromosomes with adequate follow-up information, 50% were benign including four sex chromosome markers and four autosomal markers. Two thirds of de novo marker chromosomes were associated with abnormal outcomes, while all inherited ones were benign regardless of their parental origin. Our study demonstrated that molecular characterization of prenatal marker chromosomes is of great significance in facilitating phenotype-genotype correlation. PMID- 10694687 TI - Inherited interstitial deletion of chromosomes 5p and 16q without apparent phenotypic effect: further confirmation. AB - We describe two families in which an inherited interstitial deletion is present without apparent associated phenotypic abnormalities. The first deletion was discovered in a 19-year-old male with a previously diagnosed peroxisomal disorder. High-resolution chromosome analysis was interpreted as 46,XY,del(5)(p14.1p14.3). The patient's phenotypically normal mother had the same interstitial deletion. Chromosome 5p14 deletion has been reported in a three generation family without phenotypic anomalies. We hypothesize that the affected son's phenotype may be coincidental or represent unmasking of an autosomal recessive peroxisomal disorder in the deleted region. The second interstitial deletion was detected by amniocentesis for advanced maternal age. High-resolution chromosome analysis was interpreted as 46,XX,del(16)(q13q22). The same deletion was found in the healthy mother and a normal brother. The pregnancy was carried to term and resulted in the birth of a normal girl. We report these cases as further evidence that rare, unbalanced deletion of specific chromosomal regions may result in no phenotypic effect. Consequences may result from expression of an autosomal recessive disorder on the homologous chromosome. Identification of such deletions is especially important for prenatal diagnosis and genetic counselling. PMID- 10694688 TI - Prenatal diagnosis and management of orofacial clefts PMID- 10694689 TI - Prenatal diagnosis of a fetus with a cryptic translocation 4p;18p and Wolf Hirschhorn syndrome (WHS). AB - Wolf-Hirschhorn Syndrome (WHS) is caused by distal deletion of the short arm of chromosome 4 and is characterized by growth deficiency, mental retardation, a distinctive, 'greek-helmet' facial appearance, microcephaly, ear lobe anomalies, and sacral dimples. We report a family with a balanced chromosomal translocation 4;18(p15.32;p11.21) in the father and an unbalanced translocation resulting in partial monosomy 4 and partial trisomy 18 in one living boy and a prenatally diagnosed male fetus. Both showed abnormalities consistent with WHS and had in addition aplasia of one umbilical artery. Karyotyping of another stillborn fetus revealed a supernumerary derivative chromosome der(18)t(4;18)(p15.32;p11.21) of paternal origin and two normal chromosomes 4. The umbilical cord had three normal vessels. A third stillborn fetus with the same balanced translocation as the father had a single umbilical artery and hygroma colli. PMID- 10694690 TI - Dicentric marker derived from chromosome 22 associated with mild clinical signs: a case report. AB - Amniocentesis performed after 24 weeks' gestation following ultrasonographic diagnosis of isolated unilateral hydronephrosis showed a de novo extra structurally abnormal chromosome in all cells examined. A combination of conventional and molecular cytogenetic techniques characterized the supernumerary marker as a dicentric and bisatellited marker derived from chromosome 22. At birth the infant presented hypoplasia of the right kidney, hearing loss on the left side and bilateral preauricular pits and skin tags. At three years, growth and neurological development were normal. PMID- 10694691 TI - Relative efficiency of FISH on metaphase and interphase nuclei from non-mosaic trisomic or triploid fibroblast cultures. AB - Chromosome specific probes that are used in interphase fluorescence in situ hybridization (FISH) analysis are usually tested on disomic control samples. When used for preimplantation or prenatal diagnosis the aim is to detect aneuploidy, most frequently trisomy. In this study, skin fibroblast cultures from non-mosaic trisomic and triploid fetuses were analysed by FISH to assess probe efficiency regarding interphase detection of trisomy. Skin fibroblast cultures were used because they are considered to be stable in culture. FISH experiments were performed using centromeric probes for chromosomes X, Y, 18 and locus specific probes for chromosomes 13 and 21. In metaphase nuclei, the expected signals were found in 100% of at least 30 metaphases counted on each sample and this also confirmed non-mosaicism in agreement with conventional karyotyping of the fetuses. On interphase nuclei, however, only 80-89% of nuclei per population displayed the expected signals for autosomal probes and 90% for probes for the sex chromosomes. For each probe, a range of percentages was obtained that can be regarded as indicative of non-mosaic trisomy in uncultured specimens. In the case of prenatal samples, the expected presence of maternal cells may lead to a lowering of the threshold for a trisomic diagnosis. In the case of preimplantation diagnosis, the accuracy can be improved by the use of two probes per chromosome or by the analysis of two cells from each embryo. PMID- 10694692 TI - Prenatal diagnosis of abdominal enteric duplications. AB - Alimentary tract duplications are rare congenital malformations with few reports of the antenatal sonographic appearance. Early diagnosis is of paramount importance to prevent complications. We present a case of a cystic gastric duplication diagnosed antenatally at 31 weeks' gestation, which was treated successfully. Simultaneously, we review all published cases of prenatally diagnosed enteric duplications. PMID- 10694693 TI - Plasma measurement of 7-dehydrocholesterol to detect carriers of Smith-Lemli Opitz syndrome. PMID- 10694694 TI - Maternal age-specific rates of Down syndrome used in serum screening are biased low. PMID- 10694695 TI - A false positive diagnosis of conjoined twins in a triplet pregnancy: pitfalls of first trimester ultrasonographic prenatal diagnosis. PMID- 10694696 TI - Current awareness in prenatal diagnosis. PMID- 10694697 TI - Measurement of N-methyl-2-pyridone-5-carboxamide in urine by high performance liquid chromatography. AB - We have previously described a simple and reproducible method for the measurement of nicotinamide and its major metabolite N-methyl-2-pyridone-5-carboxamide (2 pyr) in human plasma. We now describe a low-cost high-throughput method for measurement of urinary 2-pyr, and demonstrate that Isolute C18 bulk can replace use of the column to clean up the samples prior to injection into the HPLC apparatus. Using a standard curve together with an internal standard for each sample, with mean recovery of 2-pyr greater than 95%, the assay has proved reproducible, with considerable savings in cost and time. The principal advantages of this method are the rapid column clean up of samples prior to injection and the simple but effective methodology. PMID- 10694698 TI - Determination of saccharides in sake by high-performance liquid chromatography with polarized photometric detection. AB - A high-performance liquid chromatographic method has been developed for the determination of saccharides in sake, an alcoholic beverage brewed from rice. Saccharides in sake were separated on a normal phase (carbamoyl bonded silica) column using a linear gradient elution of water in acetonitrile. Seven saccharides, glucose, maltose, isomaltose, maltotriose, panose, isomaltotriose and ethyl alpha-D-glucoside, were determined by a polarized photometric detector. Unidentified peaks suggesting saccharides with polymerization degrees over 4 were also observed. The proposed method did not require any sample clean-up treatment. As an application, saccharide compositions in various kinds of sake were compared. PMID- 10694699 TI - Development of a sensitive liquid chromatography-electrospray ionization mass spectrometry method for the measurement of KW-5139 in rat plasma. AB - A sensitive method for the determination of a prokinetic peptide, KW-5139 (Leu(13)-motilin), in rat plasma has been developed utilizing liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). KW-5139 was separated by reversed-phase HPLC, with a mixture of 75 mM ammonium formate (pH 3.0) and acetonitrile (4:1, v/v), and monitored by single ion recording (SIR)-ESI MS at m/z 894 ([M+3H](3+)). Simple protein precipitation and the LC-ESI-MS analysis allowed the determination of KW-5139 in rat plasma with the mean precision and accuracy at the lower limit of quantitation (LLOQ, 0.5 ng/mL) of 5.7 and 11.2%, respectively. The method was applied to the monitoring of the plasma time-concentration profile of KW-5139, intravenously administered to rats at a dose of 1 microg/kg. PMID- 10694700 TI - HPLC-fluorescence determination of equilin and equilenin in postmenopausal women's urine. AB - A high-performance liquid chromatographic (HPLC) method with fluorescence detection (lambda(ex) = 280 nm; lambda(em) = 410 and 312 nm) in combination with a post-column on-line photochemical derivatization is described for the determination of equilin and equilenin in urine from normal postmenopausal women after therapy with conjugated oestrogens. The column effluents were subjected on line to UV irradiation (254 nm) and the photo-induced modifications were useful for the identification of the analytes. The conjugated (sulphate and glucuronide) forms were analysed after enzymatic or chemical hydrolysis and extracted with chloroform. Solid-phase extraction using strong anion-exchange sorbent was applied to the analysis of unconjugated oestrogen fraction to obtain a practical and reliable sample clean-up. The HPLC separations were achieved using ODS columns with a mobile phase consisting of 0.05 M triethylamine phosphate buffer (pH 4.0)-acetonitrile (64:36, v/v) at a flow rate of 1.0 mL/min. The method was accurate and reproducible; for the equilin and equilenin separation isocratic conditions were satisfactory, allowing a sensitive detection in urine samples with a detection limit of about 50 fmol for equilin (lambda(ex) = 280 nm; lambda(em) = 312 nm, after photoderivatization) and 10 fmol for equilenin (lambda(ex) = 280 nm; lambda(em) = 410 nm). PMID- 10694701 TI - Development of a method for the determination of danofloxacin in plasma by HPLC with fluorescence detection. AB - A simple and sensitive HPLC method has been developed for the determination of danofloxacin (DAN) in plasma. Sample preparations were carried out by adding phosphate buffer (pH 7.4, 0.1 M), followed by extraction with trichloromethane. DAN and the internal standard, sarafloxacin (SAR), were separated on a reversed phase column, and eluted with aqueous solution-acetonitrile (80:20 v/v). The fluorescence of the column effluent was monitored at lambda(ex) = 338 and lambda(em) = 425 nm. The retention times were 2.80 and 4. 40 min for DAN and SAR, respectively. The method was shown to be linear from 1 to 1500 ng/mL (r(2) = 0.999). The detection and quantitation limit were 1 and 5 ng/mL, respectively. Mean recovery was determined as 80% by the analysis of plasma standards containing 150, 750 and 1500 ng/mL. Inter- and intra-assay precisions were 4.0% and 2.4%, respectively. PMID- 10694702 TI - A rapid and sensitive method for the quantification of ganciclovir in plasma using liquid chromatography/selected reaction monitoring/mass spectrometry. AB - A method using reversed-phase liquid chromatography coupled with electrospray ionization and selected reaction monitoring mass spectrometry has been developed for the quantitative analysis of ganciclovir in rat plasma. Acyclovir, a structurally related analog of ganciclovir, was used as the internal standard. A small volume of plasma (50 microL) was spiked with the internal standard and plasma proteins were precipitated by methanol. The supernatant was dried under nitrogen, and then reconstituted in water. The use of liquid chromatography/selected reaction monitoring/mass spectrometry effectively eliminated potential interference from endogenous constituents in the plasma. This highly selective and sensitive method made it possible to analyze plasma ganciclovir with a lower limit of quantitation of 10 ng/mL. The assay was reproducible and linear in the range 10-10,000 ng/mL. The precision and accuracy values were in the range 2.0-6.9% and 89.0-109.6%, respectively. The analyte recovery was greater than 88%. This method was successfully used to monitor the pharmacokinetic profile of ganciclovir in normal rats following intraperitoneal administration of the drug. PMID- 10694703 TI - Comparative analytical quantitation of clenbuterol in biological matrices using GC-MS and EIA. AB - A simple and sensitive procedure utilizing GC-MS for the identification and quantitation of clenbuterol in biofluids and tissues is described. This improved method utilizes trimethylboroxine for the derivatization of clenbuterol, requires only 1 mL/g of biological sample, and most importantly does not require an extra cleaning step for urine specimens prior to extraction. Linear quantitative response curves have been generated for derivatized clenbuterol over a concentration range of 5-200 ng/mL. The extraction efficiency at four representative points of the standard curve exceeded 90% in both specimen types (plasma and urine). Linear regression analyses of the standard curve in both specimen types exhibited correlation coefficients ranging from 0.997 to 1.000. The Limit of detection (LOD) and Limit of quantitation (LOQ) values for plasma specimens were determined to be 0.5 and 1.5 ng/mL respectively. For urine specimens, LOD and LOQ values were 0.2 and 0.7 ng/microL respectively. Percentage recoveries ranged from 91 to 95% for urine and 89 to 101% for plasma. Precision and accuracy (within-run and between-run) studies reflected a high level of reliability and reproducibility of the method. In addition to its reliability, sensitivity and simplicity, this modified procedure is more efficient and cost effective, requiring less time, only 1 mL of sample, and minimal amounts of extraction solvents. The applicability of the method for the detection and quantitation of clenbuterol in biological tissues of rats treated with the drug was demonstrated successfully. For comparative analysis of clenbuterol in plasma and liver samples, both GC-MS and enzyme immunoassay (EIA) methods are found to be suitable. Due to potential antibody-cross reactivity with EIA, the GC-MS method is the method of choice for most samples because of its specificity. However, the EIA method is considered the method of choice for analysis of clenbuterol found in concentrations below the limits of quantitation by GC-MS due to its sensitivity. PMID- 10694704 TI - Simultaneous quantitation of d7-nefazodone, nefazodone, d7-hydroxynefazodone, hydroxynefazodone, m-chlorophenylpiperazine and triazole-dione in human plasma by liquid chromatographic-mass spectrometry. AB - A rapid and sensitive LC-MS assay was developed and validated for the simultaneous determination of d7-nefazodone (d7-NEF), nefazodone (NEF), d7 hydroxynefazodone (d7-OH-NEF), hydroxynefazodone (OH-NEF), m chlorophenylpiperazine (mCPP), and triazole-dione (Dione) in human plasma using trazodone (TRZ) as the internal standard (IS). A 0.1 mL aliquot of the plasma sample was precipitated with 0.1 mL of acetonitrile and vortexed for 2 min. After centrifugation, 50 microL of supernatant was mixed with 100 microL of 10 mM ammonium formate (pH = 4.0), and a 50 microL aliquot was injected onto a BDS Hypersil C18 column at a flow rate of 0.3 mL/min. The mobile phase consisting of 10 mM ammonium formate (pH = 4) and acetonitrile, 55:45 v/v, was used in an isocratic system. The mass spectrometer was programmed to admit the protonated molecules at m/z 477.2 (d7-NEF), 493.3 (d7-OH-NEF), 197.0 (mCPP), 372.0 (IS), 470.4 (NEF), 458.0 (Dione) and 486.2 (OH-NEF). Standard curves were linear (r(2) >/= 0.994) over the concentration range of 4-1000 ng/mL for Dione and 2-500 ng/mL for all other analytes. The lowest standard concentrations were the lower limits of quantitation for each analyte. The mean predicted quality control concentrations for all analytes deviated by less than 14.3% from the corresponding nominal values; the intra-assay and inter-assay precisions of the assay for all analytes were within 10.5% relative standard deviation. All analytes including the internal standard were stable in the injection solvent at room temperature for at least 24 h. The extraction recovery of the various analytes ranged from 79.2 to 109.1%. The validated assay was applied to the analysis of clinical samples obtained from a human subject who simultaneously received d7-NEF and NEF orally. PMID- 10694705 TI - Determination of phenobarbital in plasma by micellar liquid chromatography. AB - A new liquid chromatographic procedure for the determination of phenobarbital in plasma samples is described. The proposed system uses a Spherisorb octadecyl silane ODS-2 C(18) analytical column, a guard column of similar characteristics, and a 0.03 M CTAB-3% 1-propanol at pH 7 mobile phase. The UV detector was set at 250 nm. Butabarbital was used as internal standard. Sample preparation only required the addition to the plasma samples of a 0.1 M SDS solution at pH 3 and centrifugation before injection into the chromatographic system. The limit of detection was 0.83 microg/mL of phenobarbital in plasma samples. The coefficients of variation were lower than 7. 5%. PMID- 10694706 TI - Sensitive determination of anandamide in rat brain utilizing a coupled-column HPLC with fluorimetric detection. AB - A fluorimetric determination method for N-arachidonoylethanolamine (anandamide) was developed using a precolumn fluorescence derivatization followed by coupled column high-performance liquid chromatography (HPLC). Anandamide extracted from the rat brain tissue was derivatized with 4-N-chloroformylmethyl-N-methylamino-7 N, N-dimethylaminosulfonyl-2,1,3-benzoxadiazole (DBD-COCl), purified by a solid phase extraction (Emporetrade mark), and assayed by the coupled-column HPLC. The HPLC consisted of phenyl (100 x 4.6 mm i.d. ) and octadecylsilica columns (250 x 4.6 mm i.d.), both connected by a six-port valve. The concentration of anandamide in rat brain was 3. 37 +/- 0.73 pmol/g with 6.47 and 3.57% of intra- and inter day precisions, respectively. Using this method, we investigated the alteration of anandamide concentration in rat brain 30 min after administration of anandamide (2 mg/kg, i.p.) to rats pretreated with or without phenylmethylsulfonyl fluoride (PMSF; 30 mg/kg, i.p.), an inhibitor of amidohydrolase. In rats pretreated with PMSF, the brain concentration of anandamide was approx. 16-fold higher than that of rats without PMSF (p < 0.01). PMID- 10694707 TI - Enantiomer separation of pharmaceuticals by capillary gas chromatography with novel chiral stationary phases. AB - New chiral stationary phases of polydimethylsiloxane anchored with (S)-(-)-t leucine derivatives were provided for use in enantiomer separation of pharmaceuticals by capillary gas chromatography. Fifteen pharmaceuticals were separated into their enantiomeric pairs by converting them into pentafluoropropionyl and heptafluorobutyryl derivatives. The separation factor and resolution obtained from the new phases were superior to those from the conventional Chirasil-Val capillary column. PMID- 10694708 TI - Determination of resibufogenin and cinobufagin in heart-protecting musk pill by HPLC. AB - A high-performance liquid chromatographic (HPLC) method for the simultaneous determination of resibufogenin and cinobufagin in traditional Chinese medicine (heart-protecting musk pill, shexiang baoxin wan in Chinese) was developed. A reversed-phase system with a Hypersil (ODS2) C(18) column and tetrahydrofuran: methanol: water (8:31:61) mobile phase was employed for the separation of resibufogenin and cinobufagin. The detection was set at 299 nm and ethinyl estradiol was chosen as the internal standard. The limit of detection was 1.5 ng for resibufogenin and 2.0 ng for cinobufagin at a signal-to-noise ratio of 4:1. It is a rapid, simple and accurate method for quantitative analysis of resibufogenin and cinobufagin in heart-protecting musk pill. PMID- 10694709 TI - Fluorescent oxidation products derived from DBD-thiocarbamoyl amino acid in the modified Edman sequencing analysis. AB - The fluorescent product obtained by the oxidation of 7-N, N-dimethylaminosulfonyl 4-(2,1,3-benzoxadiazolyl) (DBD)-thiocarbamoyl (TC)-proline with NaNO(2)/H(+) in the modified Edman sequencing procedure was identified as the corresponding thiazolyl compound, N-[(8-dimethylaminosulfonyl)thiazolo[5,4, e]benzo[2,1,3]oxadiazol-5-yl]-L-proline, formed by the attack of the sulfur atom of the thiocarbamoyl group on the benzofurazan skeleton. The reaction mechanism for the formation of the fluorescent compound from DBD-TC primary and secondary amines is also discussed. PMID- 10694710 TI - Use of leucotomy for intractable anorexia nervosa: a long-term follow-up study. AB - OBJECTIVE: We studied the long-term outcomes of intractable anorexia nervosa treated with leukotomy and specialized psychotherapy over 20 years ago. METHOD: All traceable subjects were interviewed using the Eating Disorders Examination (EDE) and the Structured Clinical Interview for DSM-III-R (SCID). They also completed questionnaires. Detailed histories were taken. RESULTS: Four of five female subjects were traced. Their cases had been severe, with failure of previous intensive psychotherapy and now with high risk of death from terminal inanition. One patient had committed suicide, whereas the others enjoyed a reasonable quality of life. Persistent core psychopathology was evident, but patients had not succumbed to weight loss. All suffered depression and anxiety related disorders, but endorsed their treatment, which had allowed sustained weight gain by release of appetitive behavior, provision of a license to change, and alleviation of phobic anxiety, allowing psychotherapeutic engagement. DISCUSSION: We argue that these outcomes are relatively favorable and would not have been possible without this latter engagement in specialist psychotherapy to address burgeoning panic at unavoidable weight gain. PMID- 10694711 TI - Using the eating disorder examination to identify the specific psychopathology of binge eating disorder. AB - OBJECTIVE: The clinical features of binge eating disorder (BED) are not well established. Therefore, a comprehensive assessment of the specific psychopathology of BED as compared to anorexia nervosa (AN) and bulimia nervosa (BN) is warranted. This comparison was the aim of the present study. METHOD: Detailed ratings from an investigator-based interview, the Eating Disorders Examination (EDE), were compared across three groups of female patients: those with BED, AN, and BN, as well as normal-weight and overweight control subjects. RESULTS: When comparing BED to AN and BN, patients with BED had lower levels of restraint, eating concerns comparable to AN patients but lower than BN patients, and weight and shape concerns comparable to BN patients but higher than AN patients. Significantly more eating disorder psychopathology was found for BED patients as compared to the overweight controls on all bar the EDE restraint subscale. On the majority of individual EDE items, BED patients' scores were similar to those of AN and BN patients, including importance of shape and weight in self-evaluation and preoccupation with shape and weight. No significant relationship was found between BED patients' degree of overweight and eating psychopathology. DISCUSSION: Our findings support the status of BED as an eating disorder and suggest that the elevated EDE scores reflect the combined impact of being objectively overweight and having disordered cognitions and behaviors about eating, shape, and weight. PMID- 10694712 TI - Subthreshold binge eating disorder. AB - OBJECTIVE: To examine the clinical features of subthreshold binge eating disorder (BED). METHOD: Participants were recruited directly from the community as part of an ongoing study of risk factors for BED. Forty-four women with subthreshold BED were compared with 44 women with BED and 44 healthy controls on demographic characteristics, body mass index (BMI), eating disorder symptomatology, and psychiatric distress. Diagnoses were established using the Eating Disorder Examination (EDE). Participants completed the EDE-Questionnaire, the Brief Symptom Inventory, and were measured and weighed. RESULTS: Adjusting for significant group differences in BMI, the two eating disorder groups did not differ significantly on measures of weight and shape concern, restraint, psychiatric distress, and history of seeking treatment for an eating or weight problem. DISCUSSION: Given the importance of diagnostic status for access to treatment, further evaluation of the severity criterion specified for BED is needed. PMID- 10694713 TI - Eating disorders: psyche or soma? AB - Speculation about the etiology of eating disorders has gone through different phases, variously favoring familial, organic, and psychosocial factors. Recent evidence has particularly contributed to our understanding of the organic view. We review the evidence for an organic contribution to the illness and present a series of cases in which organic factors were present. The cases illustrate the complex interaction between biological and psychological factors. In particular, a growth hormone-producing pituitary adenoma was discovered in a patient following successful treatment of her bulimia by psychological means alone. Etiological theories of eating disorders need to encompass both organic and psychosocial factors, allowed to interact in complex ways. Focusing exclusively on either aspect is a disservice to our patients. PMID- 10694714 TI - Perceived expressed emotion in the siblings and parents of hospitalized patients with anorexia nervosa. AB - OBJECTIVE: The present study investigated the relationship between the level of perceived Expressed Emotion (EE) of the siblings and parents of patients hospitalized with anorexia nervosa and its effect on weight gain and psychological functioning. METHOD: The Level of Expressed Emotion (LEE) Scale was administered on admission to 19 patients with anorexia nervosa who completed the LEE three times so as to identify their perceptions of their relationship with their closest age sibling, mother, and father. They were also required to complete the Eating Disorder Inventory 2 (EDI-2). Patients' closest age sibling completed the Family Attitude Scale (FAS). The patients' body mass index (BMI) was calculated 6 weeks later, and the EDI-2 readministered. RESULTS: Perceived EE was not predictive of BMI change after 6 weeks of hospitalization. A composite perceived family EE score was a significant predictor of change on the Interpersonal Distrust, Maturity Fears, and Perfectionism subscales of the EDI-2. DISCUSSION: These findings suggest that patients' perceptions of their relationships with their closest aged sibling, mother, and father are poor predictors of weight gain and improvement in psychological functioning following 6 weeks of inpatient treatment. PMID- 10694715 TI - Interrelationships between the size of the pancreas and the weight of patients with eating disorders. AB - OBJECTIVE: Starvation severely affects normal pancreatic function in children suffering from Kwashiorkor and in animals undergoing food deprivation. This study examines whether pancreatic size, as determined by ultrasound, is dependent on starvation or on eating patterns in patients with eating disorders. METHOD: In 109 inpatients with eating disorders, 86 with anorexia nervosa and 23 with bulimia nervosa, we determined the pancreatic size by means of abdominal ultrasonography before increase in weight. Twenty-four inpatients with other psychiatric disorders served as controls. Pancreatic size was defined by the maximal diameter and the length of the head, the diameter of the head at the confluence of the splenic and mesenteric veins, and the diameters of the body and tail. In 41 eating disorder patients, pancreatic size was also measured during the course of therapy and increase in weight. RESULTS: Pancreatic size correlates highly with body mass index (BMI). Counteracting actions such as purging do not seem to influence this pathophysiologic finding. Dystrophy of the pancreas is reversible in a short period of time. The increase in pancreatic size after maintenance of a normal eating pattern, however, exceeded the size expected by regression equation with an increase in the BMI. Pancreatic size seems to correlate with the actual amount of digested food. The increase in BMI is only an indicator of food intake. DISCUSSION: Pancreatic size might therefore be useful for the assessment of normalization of the eating pattern. Future research is necessary to investigate the impairment of pancreatic function resulting from dystrophy, the impact of possible pancreatic malfunction on the course of eating disorders, and the regulatory mechanisms responsible for the change of pancreatic size. PMID- 10694716 TI - Nonweight-related body image concerns among female eating-disordered patients and nonclinical controls: some preliminary observations. AB - OBJECTIVE: Eating disorders(ED) have been classically associated with a concern about body shape and size that manifests mainly as an intense fear of weight gain (DSM-IV criteria). To further examine the nature of the body image disturbance in ED, we surveyed the prevalence of nonweight-related body image concerns among ED patients and nonclinical controls. METHOD: We examined 53 women (M +/- SD age: 28.1 +/- 6.8 years) with anorexia nervosa and/or bulimia nervosa (DSM-III-R criteria) and 73 randomly selected nonclinical women (M +/- SD age: 30.2 +/- 6.6 years) from the community. The participants rated (by checking a "Yes" or "No") whether they were satisfied with the appearance of the following body regions: their skin, teeth, jaw, nose, eyes, ears, hair, and height and completed the Drive for Thinness (DT) and Body Dissatisfaction (BD) subscales of the Eating Disorders Inventory (EDI). RESULTS: The frequencies of dissatisfaction with the appearance of various physical attributes among the ED patients versus the nonclinical controls were as follows: skin: 79.2% vs. 52.1%, p =.002; teeth: 62.3% vs. 39.7%, p =. 012; jaw: 24.5% vs. 9.7%, p =.026; nose: 45.3% vs. 24.7%, p =.015; eyes: 22.6% vs. 12.3%, p =.12; ears: 20.8% vs. 2.7%, p =.001; hair: 52.8% vs. 39.7%, p =.14; and height: 28.3% vs. 13.7%, p =.04. As expected, the M +/- SD DT (EDI): 14.0 +/- 6.1 vs.3.5 +/- 4.6, p <. 0001 and the M +/- SD BD (EDI): 19.7 +/- 5.8 vs. 10.1 +/- 7.3, p <. 0001, were both higher in the ED group. Furthermore, greater dissatisfaction with nonweight-related body image was associated with higher DT and BD scores. CONCLUSION: The higher prevalence of dissatisfaction with appearance of most of the nonweight-related physical attributes is probably an indication of the core ego deficits that are often present in ED and an index of the severity of the overall body image disturbance in these patients, and not indicative of another condition (e.g., body dysmorphic disorder) as the current nosology (DSM-IV) suggests. PMID- 10694717 TI - Comparisons of body image dimensions by race/ethnicity and gender in a university population. AB - OBJECTIVE: We examined affective and cognitive components of body image related to physical appearance, weight, and health among 120 university men and women of three racial/ethnic groups: African American, European American, and Latino/a American. METHOD: Participants completed a Background Information Sheet, the Multidimensional Body-Self Relations Questionnaire (MBSRQ), the Body-Esteem Scale (BES) with additional items, and the Balanced Inventory of Desirable Responding (BIDR). We tested for effects of race/ethnicity and gender on the body image measures while controlling for age, body size, social desirability, and socioeconomic status (SES). RESULTS: African Americans reported greatest body satisfaction and least overestimation of weight. Latino/a Americans were equal to or higher than European Americans on all indices. Gender differences occurred on global body image, weight concerns, fitness, and health. There were no Gender x Race/Ethnicity interactions. DISCUSSION: This pattern of racial/ethnic and gender differences shows a need for exploring a wider range of culturally relevant body image dimensions. PMID- 10694718 TI - Disordered eating in three communities of China: a comparative study of female high school students in hong kong, Shenzhen, and rural hunan. AB - OBJECTIVE: To examine disordered eating and its psychological correlates among female high school students in three Chinese communities that lay on a gradient of socioeconomic development in China. METHOD: 796 Chinese students from Hong Kong, Shenzhen, and rural Hunan completed a demographic and weight data sheet, the Eating Attitudes Test (EAT-26), a Body Dissatisfaction Scale (BDS), the Beck Depression Inventory (BDI), and the Rosenberg Self-Esteem Scale (RSES). RESULTS: Compared to students in Hunan and to a lesser extent students in Shenzhen, students from Hong Kong were slimmer, but desired a lower body mass index (BMI), reported more body dissatisfaction, exhibited a more typical EAT-26 factor structure, scored higher on the "fat concern and dieting" factor, and constituted more EAT-26 high scorers. Multiple regression analyses indicated that BDS was the most significant predictor of fat concern at each site, but this effect was strongest in Hong Kong. Hunan students had significantly higher BDI scores but lower fat concern than Shenzhen and Hong Kong students. DISCUSSION: The consistent gradient of fat concern across the three communities gives credence to the view that societal modernization fosters disordered eating in women, possibly via the gendered social constraints that accompany it. It is also expressive of the marked socioeconomic heterogeneity within China nowadays. The predictable rising rate of eating disorders that follows global change will pose a growing public health challenge to Asian countries. PMID- 10694719 TI - Subliminal activation of abandonment- and eating-related schemata: relationship with eating disordered attitudes in a nonclinical population. AB - OBJECTIVE: Previous research has demonstrated that subliminal abandonment cues can facilitate eating behavior. It is believed that such eating is a response to the activation of specific core schemata. However, the precise nature of those schemata has not been established. This study examined whether the presentation of subliminal abandonment and food/shape cues results in the activation of abandonment-related or food-related schemata. METHOD: Eighty-two women were exposed to one of three subliminal cues- an abandonment cue ("lonely"), an appetitive cue ("hungry"), and a neutral cue ("gallery"). They subsequently completed Stroop tasks to measure activation of relevant schemata. RESULTS: Subliminal presentation of abandonment cues led to the activation of food- and shape-related schemata. In contrast, subliminal appetitive cues resulted in an activation of abandonment-related schemata. CONCLUSIONS: The results show preliminary support for a multilevel cognitive model, involving indirect links between subliminal cue type and the activation of eating-related cognitions. PMID- 10694720 TI - Seasonality of eating pathology on the eating attitudes test in a nonclinical population. AB - OBJECTIVE: To determine whether there are seasonal fluctuations in eating pathology in a nonclinical population. METHOD: The Eating Attitudes Test (EAT) was completed by 322 subjects during winter and again during summer. Summer and winter responses were compared to investigate differences in EAT total and subscale scores and for individual EAT items. Numbers of subjects fluctuating across the seasons by more than 2 SDs of the cohort's scores were identified. RESULTS: The cohort showed no significant seasonal change on EAT-40 totals, EAT 26 totals, or within the EAT subscales. There were significant (p <.025) seasonal fluctuations on four of the EAT-40 questions. For individual respondents, there was no greater likelihood of scoring significantly higher in the winter than in the summer. DISCUSSION: Clinically significant seasonal fluctuations in eating pathology on the EAT did not occur in this nonclinical population. It is debatable whether items within the EAT which show significant seasonal fluctuations should be retained or discarded. PMID- 10694721 TI - The effect of body dissatisfaction on women's perceptions of female celebrities. AB - OBJECTIVE: Research suggests that media exposure causes some women to feel heightened dissatisfaction with their body shape. This study attempts to determine which women are effected as such, by investigating how women feel about their own bodies and how this effects their perceptions of female celebrities in the media. METHOD: Undergraduate females (n = 116) were shown one accurate and six distorted photographs of thin and heavy female celebrities. Each distorted photograph made the celebrity appear thinner or heavier than actuality. Participants chose which photograph portrayed each celebrity's true body shape. Body shape concerns were measured by the Body Shape Questionnaire. RESULTS: Women concerned about their body shape judged thin celebrities as thinner than actuality, whereas unconcerned women judged them accurately. Both groups judged heavy celebrities as heavier than actuality. DISCUSSION: Results suggest certain women are effected by media exposure due to their own perception of females in the media. Prevention strategies, and the media's role in body dissatisfaction and dieting disorders, are discussed. PMID- 10694722 TI - Multi-impulsivity among bulimic patients in Japan. AB - OBJECTIVE: Studies in Western world patients suggest the possible existence of a subgroup of patients with bulimia nervosa (BN) who display multiple problems with impulsivity, such as suicidal attempts. We assessed impulsive behaviors among BN patients in Japan to discuss them crossculturally. METHOD: Impulsive behaviors in 64 BN patients were assessed and multi-impulsivity (MI) was defined according to the definition proposed by Fichter, Quadflieg, and Rief (Psychological Medicine, 24, 591-604,1994). RESULTS: Nineteen patients (30%) met the definition of MI. BN patients with MI had more severe clinical features, such as concurrent depressive and anxious symptoms, global functioning, and higher prevalence of borderline personality disorder than BN patients without MI. DISCUSSION: These results showed the similarities between BN patients with MI in Japan and those patients in the Western world in clinical and psychopathological characteristics and a life-time incidence of each impulsive behavior. These findings may suggest culturally free bases for linkage between BN and MI. PMID- 10694723 TI - Personality disorders among subjects recovered from eating disorders. AB - OBJECTIVE: Personality disorders are common in symptomatic eating disorders subjects. Because personality symptoms could be exaggerated by malnutrition or Axis I disorders, we studied women who had recovered from eating disorders for at least 1 year to see if personality disorder symptoms persisted in the well state. METHOD: Personality disorders were evaluated in 10 women recovered from anorexia nervosa (AN), 28 women recovered from bulimia nervosa (BN), and 16 women recovered from AN and BN, using the Structured Clinical Interview for DSM-III-R personality disorders. RESULTS: Fourteen of 54 subjects (26%) met the criteria for at least one personality disorder, such as self-defeating, obsessive compulsive, or borderline personality disorder. Cluster B personality disorders were closely associated with bulimic subtypes. CONCLUSIONS: While a recovery from eating disorders may have an attenuating influence on the symptoms of personality disorders, such personality disorder diagnoses persist after recovery in some recovered subjects. PMID- 10694724 TI - Electrolyte and other blood serum abnormalities in normal weight bulimia nervosa: evidence for sampling bias. AB - OBJECTIVE: Sampling bias due to research settings might be responsible for reported high prevalence rates of electrolyte and serum abnormalities in bulimia nervosa. The aim of this study was to investigate the prevalence of electrolyte and other serum abnormalities in bulimia nervosa patients with normal weight seeking treatment in a community mental health center. METHOD: Diagnostic evaluations and laboratory tests were done for a consecutive series of 31 patients meeting DSM-III-R criteria for bulimia nervosa. RESULTS: The duration and clinical severity of the eating disorder were considerable and psychiatric comorbidity was high. None of the subjects showed electrolyte abnormalities. Hypomagnesemia was found in 9.7% and hypoalbuminemia in 6.4% of the population. The severity of the abnormalities was modest. DISCUSSION: The results suggest that previous reports on electrolyte abnormalities in bulimia nervosa were affected by sampling bias. Based on the results, there is no indication to perform routinely laboratory studies in ambulatory patients with normal weight. PMID- 10694725 TI - Case reports of olanzapine treatment of anorexia nervosa. AB - Two females with severe anorexia nervosa were treated with olanzapine in open trials. Olanzapine was tried because it has caused weight gain in other patient groups. Both anorexic patients had a chronic illness and had failed multiple other treatments. Olanzapine administration was associated with weight gain and maintenance as well as reduced agitation and resistance to treatment. These case histories support further exploration of this class of drugs in anorexia nervosa. PMID- 10694726 TI - Selected papers from the Oberwolfach Conference on Medical Statistics: Mathematical Models for Diagnosis and Prognosis. 1997. PMID- 10694727 TI - Prognosis - what does the clinician associate with this notion? PMID- 10694728 TI - Prevalence-free utility-respecting summary indices of diagnostic power do not exist. AB - One would like to have a ranking method for diagnostic tests that justifiably makes no assumptions about the local 'disease mix', that is, about the pre-test probabilities of the diagnostic alternatives. Assuming the simplest possible clinical context we therefore examine the following question: can a summary index of diagnostic test performance be devised which is loyal to the expected utility paradigm and, at the same time, is incidence-free (prevalence-free)? In other words, regardless of pre-test probabilities, the index should rank rival tests in a way which agrees with how they might be ranked by expected utility calculations. From a practical point of view, the answer is negative; it is proved that such indices do exist, but they necessarily impose a very restrictive mathematical form on the utility framework and, what is worse, invariably go against what is medically appropriate by violating a boundedness requirement. PMID- 10694729 TI - Sample size considerations for the evaluation of prognostic factors in survival analysis. AB - When the role of a new prognostic factor is investigated, careful planning of an appropriate study is required. This includes an assessment of the power of the study in terms of sample sizes. An adequate analysis of the independent prognostic effect of a new factor has to be adjusted for the existing standard factors. With survival time as endpoint this will usually be done with the Cox proportional hazards model. Sample size and power formulae in survival analysis have been developed by Schoenfeld for randomized treatment comparisons. In the analysis of prognostic factors the covariates included are expected to be correlated with the factor of primary interest. In this situation, the existing sample size and power formulae are not valid and may not be applied. In this paper, Schoenfeld's formula is first extended to the situation where a correlated factor is included in the analysis. The validity of the resulting approximate asymptotic formula is investigated for its asymptotic behaviour by numerical integration and for its finite behaviour by simulation. Second, an approximate formula for sample size and power is provided to detect an interaction between the interesting and a second correlated factor. This extends the formula for independent effects. Finally, the approach is illustrated by an example on the prognostic impact of DNA ploidy and other factors in advanced ovarian cancer. PMID- 10694730 TI - What do we mean by validating a prognostic model? AB - Prognostic models are used in medicine for investigating patient outcome in relation to patient and disease characteristics. Such models do not always work well in practice, so it is widely recommended that they need to be validated. The idea of validating a prognostic model is generally taken to mean establishing that it works satisfactorily for patients other than those from whose data it was derived. In this paper we examine what is meant by validation and review why it is necessary. We consider how to validate a model and suggest that it is desirable to consider two rather different aspects - statistical and clinical validity - and examine some general approaches to validation. We illustrate the issues using several case studies. PMID- 10694731 TI - Tree stability diagnostics and some remedies for instability. AB - Stability aspects of recursive partitioning procedures are investigated. Using resampling techniques, diagnostic tools to assess single split stability and overall tree stability are introduced. To correct for the procedure's preference for covariates with many unique realizations, corrected p-values are used in the factor selection component of the algorithm. Finally, methods to stabilize tree based predictors are discussed. PMID- 10694732 TI - Regional confidence bands for ROC curves. AB - The performance of a diagnostic test is characterised by its specificity and sensitivity. For a quantitative diagnostic test these criteria depend on the selected cut-off point. The receiver operating characteristic (ROC) curve of a quantitative diagnostic test is generated by plotting sensitivity against specificity as the cut-off point runs through the whole range of possible test values. In practice, the ROC curve is estimated from clinical data. One important goal is to select an optimal cut-off point. For this purpose the sample variability has to be taken into account. Recently, Campbell has introduced nonparametric asymptotic simultaneous confidence bands that are valid for the whole ROC curve. In this paper a nonparametric asymptotic approach for the construction of regional confidence bands for ROC curves is proposed. It can be applied for any specificity interval of interest. Our approach is based on the asymptotic theory of empirical and quantile processes. To investigate the small sample properties of the different approaches, a Monte Carlo study was conducted using normal and log-normal data. A method for sample size calculation is presented. Finally, the approaches are applied to a tumour marker in the diagnosis of bone marrow metastases. PMID- 10694733 TI - Comparison of diagnostic markers with repeated measurements: a non-parametric ROC curve approach. AB - In this paper we study a class of non-parametric statistics for comparing diagnostic markers with repeated measurements. Using adapted definitions of specificity and sensitivity, we suggest methods to compare the average of sensitivities across all specificities or a range of specificities. The theory allows for correlations introduced by the fact that markers may be obtained from the same patient at multiple visits and that both markers being compared may be obtained from the same patient. Results of the Monte Carlo simulations and an example from a breast cancer setting are provided. PMID- 10694734 TI - Statistical methods for the evaluation of diagnostic measurements concerning paired organs. AB - Diagnostic measurements involving paired organs may be modelled using marginal or conditional approaches. Relevant information could be ignored by using marginal models with a mean structure not including information from the fellow organ. The conditional Rosner model can be generalized to a symmetric polychotomous logistic regression model. The rather strong assumptions of the Rosner model are relaxed in this symmetric model. The use of the symmetric model is justified by its relation to discriminant analysis. Additionally, the information of the diagnostic measurements from the fellow organ is taken into account explicitly. Furthermore, in case-control studies this approach corresponds to the sampling scheme. When the status of the fellow organ and the diagnostic measurement of this organ both are available, a surprising effect may be observed; pathological values of the diagnostic measurement from the fellow organ decrease the disease probability of the selected organ. As an example, four different diagnostic procedures from the Erlangen Glaucoma Registry are investigated. PMID- 10694735 TI - On the misuses of artificial neural networks for prognostic and diagnostic classification in oncology. AB - The application of artificial neural networks (ANNs) for prognostic and diagnostic classification in clinical medicine has become very popular. In particular, feed-forward neural networks have been used extensively, often accompanied by exaggerated statements of their potential. In this paper, the essentials of feed-forward neural networks and their statistical counterparts (that is, logistic regression models) are reviewed. We point out that the uncritical use of ANNs may lead to serious problems, such as the fitting of implausible functions to describe the probability of class membership and the underestimation of misclassification probabilities. In applications of ANNs to survival data, further difficulties arise. Finally, the results of a search in the medical literature from 1991 to 1995 on applications of ANNs in oncology and some important common mistakes are reported. It is concluded that there is no evidence so far that application of ANNs represents real progress in the field of diagnosis and prognosis in oncology. PMID- 10694737 TI - Methods of ordinal classification applied to medical scoring systems. AB - Scoring systems are used in nearly all fields of medicine for evaluation of the state of a disease. The prediction performance of scoring systems with respect to an ordinal outcome scale is investigated, based on grouped continuous logistic models as well as on an extension of the stereotype logistic regression model. The latter is a canonical approach, which allows assessment of properties of outcome categories such as partial and total ordering, distinguishability and allocatability. The approach is applied to a data set of patients with injuries of the head. PMID- 10694736 TI - Choice of conditional models in bivariate survival. AB - We consider bivariate survival problems in which interest is in the conditional distribution of one survival variable given an uncensored observation of the other. The work is motivated by an analysis of time to cancer diagnosis then subsequent survival amongst a group of organ transplant recipients. The effect of conditioning is illustrated for five standard bivariate models. The consequences of adopting a misspecified marginal approach in which the conditioning variable is considered to be a fixed covariate are investigated. PMID- 10694738 TI - Multi-state models for bleeding episodes and mortality in liver cirrhosis. AB - Data from a controlled clinical trial in liver cirrhosis are used to illustrate that multi-state models may be a useful tool in the analysis of data where survival is the ultimate outcome of interest but where intermediate, transient states are identified. We compare models for the marginal survival time distribution with models including transient states, both with respect to their clinical interpretation and with respect to the precision of survival probability estimates obtained from the various models. PMID- 10694739 TI - Identifying and modelling prognostic factors with censored data. AB - A major issue in the analysis of diseases is the identification and assessment of prognostic factors relevant to the development of the illness. Statistical analyses within the proportional hazards framework suffer from a lack of flexibility due to stringent model assumptions such as additivity and time constancy of effects. In this paper we use tree-based models and varying coefficient models to allow for detectability of prognostic factors with possibly non-additive, non-linear and time-varying impact on disease development. Questions concerning model and smoothing parameter selection are addressed. An analysis of a data set of breast cancer patients demonstrates the ability of these methods to reveal additional insight into the disease influencing mechanisms. PMID- 10694740 TI - Statistical models for longitudinal biomarkers of disease onset. AB - We consider the analysis of serial biomarkers to screen and monitor individuals in a given population for onset of a specific disease of interest. The biomarker readings are subject to error. We survey some of the existing literature and concentrate on two recently proposed models. The first is a fully Bayesian hierarchical structure for a mixed effects segmented regression model. Posterior estimates of the changepoint (onset time) distribution are obtained by Gibbs sampling. The second is a hidden changepoint model in which the onset time distribution is estimated by maximum likelihood using the EM algorithm. Both methods lead to a dynamic index that represents a strength of evidence that onset has occurred by the current time in an individual subject. The methods are applied to some large data sets concerning prostate specific antigen (PSA) as a serial marker for prostate cancer. Rules based on the indices are compared to standard diagnostic criteria through the use of ROC curves adapted for longitudinal data. PMID- 10694741 TI - Identification of monoclonal nonspecific suppressor factor beta (mNSFbeta) as one of the genes differentially expressed at implantation sites compared to interimplantation sites in the mouse uterus. AB - Successful implantation requires synchronous development of and active dialogue between the maternal endometrium and the implanting blastocyst. While it is well established that appropriate maternal steroid hormones are essential for endometrial preparation for implantation, the molecular events at the actual site of implantation are still little understood. The aims of our studies were to identify genes explicitly expressed or repressed at the sites of implantation by utilising RNA differential display (DDPCR), and to establish the roles of these genes in the implantation process in a mouse model. Ten bands unique in implantation sites compared to interimplantation sites were identified by DDPCR and subsequently confirmed by Northern blotting. One of these bands contained a cDNA fragment that was highly homologous to mouse monoclonal nonspecific suppressor factor beta (MNSFbeta) or Fau. The full cDNA sequence of this gene, obtained by screening a lambdagt11 cDNA library, was essentially the same as MNSFbeta, except that it had much longer 5' untranslated region. Interestingly, both Northern and immunohistochemical analysis showed that the expression of this gene was much lower in implantation sites compared to interimplantation sites on day 4.5 of pregnancy, when embryos first attach to the uterus and initiate implantation, and on day 5.5, when implantation has advanced. These results suggest a role for MNSF during implantation and early pregnancy, possibly through regulating the proliferation and/or differentiation of uterine stromal cells. It may also be involved in the selective production of TH2-type cytokines in implantation sites to regulate the immune system at the maternal-fetal interface. PMID- 10694742 TI - Cloning and characterization of a vasa-like gene in rainbow trout and its expression in the germ cell lineage. AB - The origin of germ cells and the molecular mechanisms of primordial germ cell (PGC) determination in teleosts are unclear. Vasa is a member of the DEAD protein family and plays an indispensable role in germ cell determination in Drosophila and Xenopus species. In this study, we isolated and characterized a rainbow trout vasa cDNA as a first step towards understanding the molecular mechanisms of PGC determination and development and to develop a molecular marker to identify the PGCs in rainbow trout. Cloning of vasa cDNA was performed by degenerate- and RACE PCR. The predicted amino acid sequence of rainbow trout Vasa contained eight consensus sequences for the DEAD protein family and five arginine-glycine-glycine repeats, a common character of known Vasa homologues. Overall amino acid similarity to the Vasa of Drosophila was 79.2%. Whole-mount in situ hybridization of eyed stage embryos (eighty somite stage) revealed that signals were localized to the putative PGCs. In adult rainbow trout tissues, both ovaries and testes contained large amounts of vasa gene transcripts. A reverse transcription polymerase chain reaction analysis of unfertilized eggs proved that trout vasa is a maternal factor. Although we have not determined whether rainbow trout vasa functions as a germ cell determinant, its limited expression in the germ cell lineage proved that rainbow trout vasa can be used as a marker molecule for PGCs. This marker will make it possible to identify the PGCs or presumptive PGCs in early trout embryos whose germ cells can not be distinguished by morphological characteristics. PMID- 10694743 TI - Murine Myak, a member of a family of yeast YAK1-related genes, is highly expressed in hormonally modulated epithelia in the reproductive system and in the embryonic central nervous system. AB - We have cloned a mouse homologue (designated Myak) of the yeast protein kinase YAK1. The 1210 aa open reading frame contains a putative protein kinase domain, nuclear localization sequences and PEST sequences. Myak appears to be a member of a growing family of YAK1-related genes that include Drosophila and human Minibrain as well as a recently identified rat gene ANPK that encode a steroid hormone receptor interacting protein. RNA blot analysis revealed that Myak is expressed at low levels ubiquitously but at high levels in reproductive tissues, including testis, epididymis, ovary, uterus, and mammary gland, as well as in brain and kidney. In situ hybridization analysis on selected tissues revealed that Myak is particularly abundant in the hormonally modulated epithelia of the epididymis, mammary gland, and uterus, in round spermatids in the testis, and in the corpora lutea in the ovary. Myak is also highly expressed in the aqueduct of the adult brain and in the brain and spinal cord of day 12.5 embryos. PMID- 10694745 TI - LiCl disrupts axial development in mouse but does not act through the beta catenin/Lef-1 pathway. AB - Chimera and cell marking studies suggest that axial determination in mouse embryos occurs at postimplantation stages. In contrast, Xenopus laevis axes are determined early due to the asymmetric distribution of maternally derived factors in the one-cell zygote. In our earlier study we used lithium chloride (LiCl) to perturb development of mouse axes. Here we investigate whether the lithium induced axial defects in mouse are being mediated by the beta-catenin/Lef-1 pathway as in Xenopus laevis. In lithium treated embryos we did not observe any changes in the amount or localization of beta-catenin protein. Furthermore, the lack of Lef-1 mRNA in treated and untreated embryos indicates the LiCl induced axial defects in the mouse are not mediated by the beta-catenin/Lef-1 pathway. PMID- 10694744 TI - Expression of pituitary adenylate cyclase activating polypeptide (PACAP) and PACAP type I A receptor mRNAs in granulosa cells of preovulatory follicles of the rat ovary. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) was isolated from ovine hypothalamus and known to stimulate the production of cAMP in anterior pituitary cells. In the recent report, the expression of PACAP was detected in preovulatory follicles, and treatment with PACAP stimulated the production of progesterone and prostaglandin E(2) through the action of AC and PLC pathways in the ovary. PACAP binds to three type receptors. Type I A receptor is coupled to adenylate cyclase (AC) and phospholipase C (PLC) pathways, while type I B and type II receptors are only coupled to AC. Thus, the present study aimed to evaluate the temporal expression of PACAP and its type I A receptor mRNAs in the rat ovary after treatment with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG). Northern blot analysis showed that PACAP transcripts were transiently expressed from 3-9 hr after hCG treatment, reaching a maximum at 6 hr. During these time points, PACAP mRNAs were specifically and strongly expressed in granulosa cells and cumulus cells of large preovulatory follicles and interstitial glandular cells. Type I A receptor mRNAs were also transiently expressed in granulosa cells of large preovulatory follicles from 3-9 hr after hCG treatment. PACAP and its type I A receptor mRNAs were expressed in the same preovulatory follicles. These results demonstrate that PACAP acts as an autoregulator or pararegulator through type I A receptor in granulosa cells and cumulus cells of large preovulatory follicles. Thus, we suggest that PACAP may have a critical role in granulosa cells of preovulatory follicles for the preparation of ovulation. PMID- 10694746 TI - Relationship of gap junction formation to phosphorylation of connexin43 in mouse preimplantation embryos. AB - To clarify the relationship of gap junction formation to phosphorylation of connexin43 (Cx43) in mouse preimplantation embryos, immunofluorescence and Western blot analysis were conducted. Immunofluorescence showed Cx43 positive spots first at the mid-eight-cell stage (6 hr postdivision to the eight-cell stage). The number of spots increased from 6 to 15 hr postdivision to the eight cell stage. Western blot analysis suggested Cx43 to possibly be present in the nonphosphorylated form at the mid-four-cell stage (6 hr postdivision to the four cell stage), and phosphorylated Cx43 to increase from the mid-eight-cell stage (6 hr post-division to the eight-cell stage) onward. Dibutyryl cAMP (dbcAMP), a protein kinase A (PKA) activator, added to the culture medium increased the phosphorylation of Cx43 and Cx43 positive spots. The tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC) activator, increased the phosphorylation of Cx43, but decreased Cx43 positive spots. These results suggest that the phosphorylation of Cx43, induced by different protein kinase, leads to a different effect on gap junction formation in mouse preimplantation embryos. PMID- 10694747 TI - Apoptosis of fetal testicular cells is regulated by both p53-dependent and independent mechanisms. AB - A portion of fetal germ cells undergoes apoptosis in the physiological context, but the molecular mechanisms of their apoptosis are largely unknown. Because p53 tumor suppressor gene product promotes apoptosis in various types of cells, we have investigated the expression of p53 in fetal gonads and examined the influence of loss of p53 function in fetal gonad cells using mice deficient in the p53 gene. We found that the expression of p53 protein in fetal testis was induced after 15.5 dpc (days post coitum), while the expression was not detected in fetal ovary. The number of apoptotic cells found in the seminiferous tubules of fetal testes was not significantly different between p53-deficient and wild type mice until 16.5 dpc. At 17.5 dpc, however, more apoptotic cells were observed in wild-type testes than in the p53-deficient mice. In contrast, a similar number of apoptotic cells was found in fetal ovaries throughout these developmental stages. These observations indicate that p53 promotes apoptosis of fetal testicular cells after 16.5 dpc. PMID- 10694748 TI - Tumor necrosis factor-alpha and its receptor in the corpus luteum of pregnant cows. AB - The objective of this study was to investigate the presence of tumor necrosis factor (TNF)-alpha mRNA and TNF-alpha receptors in the bovine corpus luteum (CL) during the gestation period. TNF-alpha mRNA and TNF-alpha receptors were determined on bovine CL from pregnant cows at three stages: trimester I (fetal crown-rump length; 6-20 cm), trimester II (25-45 cm) and trimester III (50-80 cm). TNF-alpha mRNA was detected by an RT-PCR analysis in the CL of all stages of gestation. A Scatchard analysis revealed the presence of a high-affinity binding site (Kd; 5.1-6.9 nM) in the CL membranes collected at each stage of gestation. Furthermore, the concentrations of TNF-alpha receptors in the CL of trimesters I (24. 0 +/- 1.95 pmol/mg protein) and III (21.6 +/- 2.39 pmol/mg protein) of gestation were significantly higher than the concentration in trimester II (14.9 +/- 2.07 pmol/mg protein) (P < 0.05). These results indicate that TNF-alpha is locally produced and that TNF-alpha receptors are present in bovine CL during the gestation period, and suggest that TNF-alpha plays one or more roles as a paracrine factor in regulating bovine CL function during the entire gestation period. PMID- 10694749 TI - Nitric oxide is essential for optimal meiotic maturation of murine cumulus-oocyte complexes in vitro. AB - This study was conducted to determine whether endothelial-derived nitric oxide synthase (eNOS) affects meiotic maturation of mouse oocytes in vitro. Cumulus oocyte complexes (COC) were isolated from ovarian follicles of 27-day-old PMSG primed wildtype (WT), and eNOS-knockout (eNOS-KO) females, and cultured in drops of medium under oil at 37 degrees C for 16-18 hr. Experiment 1 was carried out to determine effects of eNOS deficiency on the ability of COC to mature in vitro. To determine whether acute synthesis of nitric oxide (NO) was required for oocyte maturation, COC collected from WT mice were cultured in medium without (control) or with different doses of N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NOS (exp. 2). To assess effects of NO deficiency on the kinetics of germinal vesicle breakdown (GVBD), COC from WT and eNOS-KO females were observed for 3.5 hr. COC from WT females were also incubated in medium without or with L NAME (exp. 3 and 4). After the culture period, cumulus cells were removed, and oocytes were counted and classified as metaphase II (M II), metaphase I (M I) or showing atypical (degenerative) morphology. To determine viability and nuclear morphology of oocytes, they were stained with fluorescein diacetate or 4,6 diamidine-2'-phenylindole dihydrochloride, respectively. There were no differences in body weights but ovarian weights were lower in eNOS-KO mice compared with WT mice (P < 0.05). Ovaries from eNOS-KO mice contained fewer COC collected relative to WT mice (P < 0.01). Maturation of COC from eNOS-KO mice or WT oocytes treated with L-NAME resulted in a lower percentage of oocytes at M II stage (P < 0.01 and P < 0.05, respectively) and a higher percentage of oocytes at M I or atypical stages compared with those from WT (P < 0.01 and P < 0.05, respectively). Many oocytes that showed either an arrest in M I stage or abnormal morphology were not viable. Several oocytes in M II stage demonstrated abnormalities in distribution of maternal chromosomes. Our data demonstrate that eNOS-derived NO is a key modulator of oocyte meiotic maturation in vitro. These results support our previous observations in vivo and indicate that eNOS/NO has independent functions in both oocyte maturation and follicular/oocyte development. PMID- 10694750 TI - Activation of bovine oocytes by specific inhibition of cyclin-dependent kinases. AB - Activation of bovine oocytes by experimental procedures that closely mimic normal fertilization and allow to obtain haploid oocytes is essential both for intracytoplasmic sperm injection (ICSI) and for nuclear transfer. Therefore, with the goal of producing haploid activated oocytes, this study evaluated whether bohemine, either alone or in combination with ionomycin, is able to activate young matured bovine oocytes. Furthermore, the effect of bohemine on the patterns of DNA synthesis after pronuclear formation as well as changes in histone H1 kinase and MAP kinase activities during the process of activation were studied. Our results with bohemine show that the specific inhibition of CDKs in metaphase II bovine oocytes induces parthenogenetic activation in a dose-dependent manner (25, 50, and 100 microM, respectively), either alone (3%, 30%, and 50%) or in combination with ionomycin (30%, 70%, and 87.5%). A single pronucleus and extrusion of the second polar body was observed (97%) when Ca(2+) influx was stimulated in the presence of bohemine, although pronuclear formation without polar body extrusion was observed when bohemine was used alone. Bohemine activated oocytes started to synthesize DNA in the first hour (37%) after their removal from bohemine-supplemented medium (6-7 hr post-activation; hpa). A high synchrony in the S-phase was registered with more than 85% of parthenotes actively synthesizing DNA 8 hpa. By contrast, DNA synthesis was absent in oocytes cultured for 4, 6, and 8 hpa in the presence of bohemine and a low rate was observed by those cultured for 18 hr (30%) in bohemine-supplemented medium. This confirms the ability of the inhibitor to arrest the cell cycle in the G1/S boundary for at least 8 hr. A drop in histone H1 kinase activity was observed in bohemine-activated oocytes. The activity of MBP kinase decreased later than histone H1 kinase and even 4 hr after inomycin-bohemine treatment at least half of this activity was still detectable. Then, the MBP kinase activity decreased and the lowest level could be seen 6-8 hpa. In summary, our study shows that in vitro matured bovine oocytes can be successfully activated by a synthetic inhibitor of CDKs. This effect can be improved by combination with ionomycin. The targeting of CDKs in the way to activate bovine oocytes can be an approach to improve the efficiency of mammalian oocyte activation. PMID- 10694751 TI - Bovine cumulus/oocyte complex: quantification of LH/hCG receptors. AB - LH/hCG receptors of the bovine cumulus/oocyte complex were quantified, and their maximum binding capacities and affinity constants were determined by Scatchard analysis. Specific binding of these gonadotropins to receptors in follicles of different sizes was also determined by radiolabeling techniques. A greater number of receptors was observed to be bound to LH than to hCG (P < 0.05); however, affinity constants did not significantly differ (P > 0.05). The results of specific binding of the gonadotropins presented differences in relation to follicle size. Differences in the specific binding values of LH and hCG were verified (P < 0.05), but when submitted to linear regression analysis, presented similar behaviors in relation to follicle size. It is concluded that receptors of bovine cumulus/oocyte complex cells bind specifically to LH/hCG, that binding capacity is inversely proportional to follicle size, and that the behavior of hCG is similar to that of LH, suggesting that hCG can also promote the maturation of bovine oocytes when used in concentrations greater than LH. PMID- 10694752 TI - Effects of ethanol and dimethylsulphoxide on nuclear and cytoplasmic maturation of bovine cumulus-oocyte complexes. AB - The influence of small doses of ethanol or dimethylsulphoxide (DMSO) on in vitro maturation (IVM) of bovine cumulus-oocyte complexes (COC) was examined, either after spontanous maturation or after inhibition of meiosis with 6 dimethylaminopurine (6-DMAP) or 3-isobutyl-1-methylxanthine (IBMX). Subsequent to IVM for 23 hr in semidefined serum-free Earle's TCM199 medium, nuclear maturation was assessed cytogenetically, while the combined cytoplasmic and nuclear maturation was measured indirectly by the oocytes' ability to undergo fertilization and further embryonic development. Embryo development was followed until the blastocyst stages at day 9 after insemination. Neither spontanous nuclear maturation nor cleavage was compromised by IVM in 0.25). Second, individual porcine and bovine sperm plasma membrane proteins were identified by binding of homologous or heterologous oocyte plasma membrane to whole sperm plasma membrane on Western ligand blots. The relative amount of labeled oocyte plasma membrane bound to individual sperm plasma membrane proteins was analyzed by laser densitometry. Eight porcine sperm plasma membrane proteins and seven bovine sperm plasma membrane proteins were bound by both porcine and bovine oocyte plasma membrane. A significantly greater relative amount of porcine oocyte plasma membrane than bovine oocyte plasma membrane was bound to the 14- and 10-kD porcine sperm plasma membrane proteins (P < 0.001 and P < 0.01, respectively). A 27-kD bovine sperm plasma membrane protein bound proportionally more bovine oocyte plasma membrane probe than porcine oocyte plasma membrane probe (P < 0.04). These results are consistent with conservation of similar receptor ligand interactions at the gamete plasma membrane among porcine and bovine gametes. PMID- 10694754 TI - Oct-4: control of totipotency and germline determination. AB - The transcription factor Oct-4 is expressed in totipotent embryonic cells and germ cells. As totipotent cells differentiate to form somatic and/or extraembryonic tissues, the Oct-4 gene is downregulated. Primordial germ cells are the only cells in which Oct-4 expression is maintained after postgastrulation. Recent in vivo ablation of the Oct-4 function has shown that the absence of this transcription factor causes early embryonic lethality due to trophectodermal differentiation of cells which normally would give rise to the inner cell mass of the blastocyst. This result strongly suggests that Oct-4 is necessary for the maintenance of the totipotent phenotype of embryonic cells and that this factor likely plays a role as a determinant of the totipotency of germ cells by preventing their differentiation to a somatic cell phenotype during gastrulation. The involvement of Oct-4 in the biology of totipotent and germ cells is here discussed in view of new understanding about Oct-4 function. PMID- 10694755 TI - Managed care in the United States: a dilemma for evidence-based policy? PMID- 10694756 TI - The relationship between marijuana initiation and dropping out of high school. AB - The prevalence of marijuana use among young people has risen rapidly in recent years, causing concern over the potential impact on academic performance of such use. While recent studies have examined the effect of alcohol use on educational attainment, they have largely ignored the potential negative effects of other substances, such as marijuana. This paper examines whether the relationship between the initiation of marijuana use and the decision to drop out of high school varies with the age of dropout or with multiple substance use. Data are from a longitudinal survey of 1392 adolescents aged 16-18 years. The results suggest that marijuana initiation is positively related to dropping out of high school. Although the magnitude and significance of this relationship varies with age of dropout and with other substances used, it is concluded that the effect of marijuana initiation on the probability of subsequent high school dropout is relatively stable, with marijuana users' odds of dropping out being about 2.3 times that of non-users. Implications of these conclusions are considered for both policy makers and researchers. PMID- 10694757 TI - Policy analysis of the use of hepatitis B, Haemophilus influenzae type b-, Streptococcus pneumoniae-conjugate and rotavirus vaccines in national immunization schedules. AB - After the development of national vaccine programmes to deliver six vaccines to infants, new vaccine adoption has been limited. Analysis of the health and economic implications of new vaccination options can help national policy-makers. Country specific quantitative policy analyses were conducted to estimate the impact of vaccination against hepatitis B (HB), Haemophilus influenzae type b (Hib), Streptococcus pneumoniae (SP) and rotavirus. Disease burden, programme costs and the potential reduction of disease from vaccination was assessed for each vaccine. Without vaccination, these four vaccine preventable diseases contribute up to 4.1 million deaths in each successive birth cohort. Routine scheduled use of HB and Hib vaccines could prevent up to 1.7 million deaths; SP and rotavirus vaccines, an additional 1.4 million deaths, annually. The global cost per life-year saved ranged from $29 to $150 with great variation by income and economic groups. With a few exceptions for a few countries, these vaccines would cost a fraction of average per-capita gross domestic product to save a life year. The addition of HB and Hib vaccines, should be considered for integration in all national immunization programmes. SP and rotavirus vaccines, with the given assumptions, would also be cost-effective. Proactive analysis of the economic and epidemiologic impact of these vaccines can hasten their introduction into national vaccination schedules. PMID- 10694758 TI - Measuring productivity loss days in asthma patients. The Pharmacy Medication Monitoring Program and Advisory Board. AB - In assessments of the cost of illness, productivity losses potentially constitute a large proportion. The present study objective was to develop a method to measure restricted days and to quantify total productivity loss days (PLDs) in adult asthma patients. Patient and disease characteristics, occupation, annual wage, work absences, restricted days, level of functioning on restricted days, and travel and waiting time were collected over 6 months in 892 adult asthma outpatients residing in southern Ontario. Annual PLDs varied from 12 in employed persons to 49 in disability pensioners. Homemakers reported 22 PLDs per year. Restricted days accounted for most PLDs and functional level during restricted days varied from 55% to 81%. Annual PLDs increased with increasing disease severity. Employed persons experienced the fewest PLDs and functioned at the highest level during restricted days, but also demonstrated a milder disease compared with other groups. Most productivity loss in asthma patients resulted from numerous restricted days, a category of PLD that is often ignored in economic assessments. The presentation of PLD results disaggregated by category of time loss and wage rate may provide valuable information to employers and health policy makers and may facilitate the application of multiple approaches to the calculation of indirect costs. PMID- 10694759 TI - Are preferences over health states complete? AB - Most applied work in health economics accepts, if only implicitly, the axiom of completeness. Preferences over health states or health services are assumed to be well formed. They are effectively 'data' waiting to be collected. An alternative perspective suggests that values are initially incomplete and are constructed rather than just revealed in the process of answering choice-related questions such as willingness to pay or standard gambles. What might appear as measurement error may, therefore, be a more deliberate process of reflection and deliberation. This paper reports on a study that assessed the completeness of health preferences. The results show a mixed pattern. For most of the sample, values were stable over repeat administration, suggesting completeness. However, one-third of participants deliberately changed their answers and suggested that the interview process had forced them to think about their values more deeply. While it is premature to draw conclusions from this small sample, the suggestion is that completeness cannot be taken for granted. PMID- 10694760 TI - QALYs and ageism: philosophical theories and age weighting. AB - QALY maximization is sometimes criticized for being 'ageist', because, other things being equal, the elderly, with a shorter life expectancy, will be given lower priority. On the other hand, there are philosophical arguments that, for different reasons, advocate rationing health care to the elderly, even when the size of the expected benefits in QALY terms is the same across older and younger patients. This paper examines six proposals, both from the philosophical and the health economics literature, that will lead to such conclusions. These are: two variants of the so-called fair innings argument, the fair innings weights, the Disability Adjusted Life Year (DALY) age weighting, the biographical life span, and the prudential lifetime account. Two questions are addressed with regard to each of these. First, what is the reason for choosing the younger patient when the QALY gains are equal; second, will the younger patient continue to be chosen even when the QALY gains to the older patient are larger. The paper studies the relationship between the six proposals and explores their possible implications for QALY maximization. PMID- 10694761 TI - Progress and directions in refining the global burden of disease approach: a response to Williams. PMID- 10694762 TI - Comments on the response by Murray and Lopez. PMID- 10694764 TI - Anaemia in pregnancy: possible causes and risk factors in Nepali women. AB - OBJECTIVE: The aim of this study was to investigate the importance of nutritional deficiencies and infections in the development of anaemia in pregnant Nepali women. DESIGN: Case-control study. SETTING: Patan Hospital, Kathmandu, Nepal. SUBJECTS: A sub-sample (n=479) of all pregnant women (n=2856) coming for their first antenatal visit in a 12 month period, 1994-1995. Women who had already received any micronutrient supplementation (n=82), and those whose serum samples showed macroscopic haemolysis (n=7) were excluded. The remaining women (n=390) were included in the statistical analysis. They were divided into three groups; a non-anaemic control group, haematocrit (Hct)>33% (n=82), and two case-groups: moderately anaemic, Hct 25-33% (n=254), and severely anaemic, Hct<25% (n=54). RESULTS: We found high prevalences of nutritional deficiencies and intestinal infections, both among cases and controls. The prevalence of low s-ferritin was high, especially among the severely anaemic women (55.6%). In a multiple logistic regression model, the presence of low s-vitamin A, elevated s-C-reactive protein or hookworm infection was associated with a significantly increased risk of severe anaemia. The adjusted odds ratios (95% CI) were 8.38 (1.99, 35.30), 4.91 (1.22, 19.67) and 5.43 (1.20, 24.61), respectively. CONCLUSIONS: In addition to the present routine iron and folate supplementation to pregnant Nepali women, vitamin A supplementation needs to be considered. Prevention and treatment of infections should, together with dietary advice, be emphasized more strongly in the antenatal care. SPONSORSHIP: The Norwegian Research Council and the Norwegian Universities Committee for Development, Research and Education. European Journal of Clinical Nutrition (2000) 54, 3-8 PMID- 10694765 TI - Relation of lifestyle factors to metacarpal bone mineral density was different depending on menstrual condition and years since menopause in Japanese women. AB - OBJECTIVE: To examine whether the relation between lifestyle and metacarpal bone mineral density (BMD) varied with life-stages. DESIGN: Cross-sectional study. SETTING: Taking questionnaires related to lifestyle and examining metacarpal BMD by computed X-ray densitometry at a local health care center. SUBJECTS: Out of 750 Japanese women aged 40-69 y old who underwent screening for osteoporosis, we selected 535 healthy subjects without medical conditions or lifestyle factors known to affect bone metabolism. RESULTS: A cross-sectional comparison of BMD and years since menopause (YSM) resulted in a logarithmic regression model (BMD=2. 539-0.149xloge YSM, r2=0.418), indicating that a prominent reduction in metacarpal BMD appears up to 4-6 y after menopause. In premenopausal women with regular menstruation, those who engaged in regular physical activity during adolescence showed significantly greater BMD than those who did not. In 1-5-y postmenopausal women, those with current calcium intake of more than 800 mg/d, current milk intake greater than 900 ml/week, daily consumption of milk and/or dairy products, or frequent consumption of small fish showed significantly greater BMD. In 6-15-y postmenopausal women, those who took more than 6000 or 8000 steps/day showed significantly greater BMD. CONCLUSIONS: The relation between lifestyle and BMD differed with life-stages in Japanese women. It was suggested that to perform lifestyle modifications for the prevention of osteoporosis, life-stages should be taken into consideration. SPONSORSHIP: Grant for Research on Health Service from Ministry of Health and Welfare, Japan and grant from The National Dairy Promotion and Research Association of Japan. European Journal of Clinical Nutrition (2000) 54, 9-13 PMID- 10694766 TI - Effects of soy as tofu vs meat on lipoprotein concentrations. AB - OBJECTIVES: To investigate the effect of replacing lean meat with a soy product, tofu, on serum lipoprotein concentrations. STUDY AND DESIGN: Randomized cross over dietary intervention study. SUBJECTS: Forty-two free-living healthy males aged 35-62 y completed the dietary intervention. Three additional subjects were non-compliant and excluded prior to analysis. INTERVENTIONS: A diet containing lean meat (150 g/d) was compared with one with 290 g/d tofu in an isocaloric and isoprotein substitution. Both diet periods were 1 month, and fat intake was carefully controlled. RESULTS: Seven-day diet records showed the two diets were similar in energy, macronutrients and fibre. Total cholesterol (mean difference 0.23 mmol/l, 95% CI 0.02, 0.43; P=0.03) and triglycerides (mean difference 0.15 mmol/l, 95% CI 0.02, 0.31; P=0.017) were significantly lower on the tofu diet than the lean meat diet. However, HDL-C was also significantly lower on the tofu diet (mean difference 0.08 mmol/l, 95% CI 0.02, 0.14; P=0.01) although the LDL C:HDL-C ratio was similar. CONCLUSION: The effect on HDL-C and the small LDL-C reduction differ from some other studies, where fat was often less controlled, and the comparison was of soy as textured protein or soymilk against casein. This suggests a differential effect of the various proteins compared to the soy may influence the findings. In practice, the replacement of meat with tofu would usually be associated with a decrease in saturated fat and an increase in polyunsaturated fat and this should enhance any small benefits due to the soy protein. SPONSOR: Deakin University with some contribution from a Commonwealth Department of Veterans Affairs research grant. European Journal of Clinical Nutrition (2000) 54, 14-19 PMID- 10694767 TI - Near infrared spectroscopy-a potentially useful method for rapid determination of fat and protein content in homogenized diets. AB - OBJECTIVE: To investigate the potential of near-infrared reflectance (NIR) spectroscopy as a rapid and non-destructive method to determine total fat and protein in mixed, homogenized and freeze-dried human diets. DESIGN: 29 students collected duplicate portions of their diet for four consecutive days. In addition, a detailed food diary was kept. The duplicate portions were analysed for total protein and fat content both by traditional chemical analysis (Kjeldahl and Folch methods) and through the recently developed NIR spectroscopy method. In addition, traditional computerized estimation of nutrient composition was performed. RESULTS: Plotting of the NIR-predicted fat content against the chemically analysed fat content gave a correlation coefficient of 0. 99. Plotting of the NIR-predicted protein content against the Kjeldahl-analysed protein gave a correlation coefficient of 0.81. CONCLUSION: NIR-spectroscopy seems to be able to determine fat content in mixed, homogenized diets to a high degree of accuracy. In surveys involving duplicate portion sampling this will save time and money. The prediction accuracy for protein was less convincing, but acceptable depending on the need for accurate individual data. SPONSORSHIP: Norwegian Food Research Institute, Institute for Nutrition Research at the University of Oslo and the Research Society of the Norwegian Edible Fat Producers and the food company Mills DA. European Journal of Clinical Nutrition (2000) 54, 20-23 PMID- 10694768 TI - Effects of a low-glycaemic index spaghetti meal on glucose tolerance and lipaemia at a subsequent meal in healthy subjects. AB - OBJECTIVE: The aim of the study was to evaluate the impact of a low glycaemic index (GI) breakfast on glucose tolerance and lipaemia at a subsequent lunch meal. DESIGN: A low GI spaghetti meal and a high-GI white wheat bread (WWB) reference meal were served in the morning after an overnight fast in random order. Four hours after the breakfast, the subjects were given a second meal-a standardized high-GI lunch-and the blood glucose, insulin and lipid responses were measured after the lunch meal. SETTING: The study was performed at the Department of Applied Nutrition and Food Chemistry, Lund University, Sweden. SUBJECTS: Ten healthy volunteers, eight women and two men, aged 25-51 y, with normal body mass indices, were recruited. RESULTS: Lowered glucose and insulin responses and reduced serum triglyceride (TG) level were found at the subsequent lunch meal when the low-GI spaghetti meal was given as a breakfast. No differences in total serum cholesterol or HDL cholesterol were seen after lunch, when preceded by the WWB reference breakfast or the spaghetti breakfast, respectively. CONCLUSIONS: Improved glucose tolerance and lowered serum TG levels can appear in the course of a single day. As insulin resistance and raised postprandial TG concentration are known risk factors for cardiovascular disease, the present study adds evidence for a beneficial role of a low-GI diet. SPONSORSHIP: Cerealia Foundation for Research and Development (project no. 232) and the Swedish Council for Forestry and Agricultural Research. European Journal of Clinical Nutrition (2000) 54, 24-28 PMID- 10694769 TI - Effect of daily iron supplementation on iron status, cell-mediated immunity, and incidence of infections in 6-36 month old Togolese children. AB - OBJECTIVE: To assess the impact of a daily oral iron supplementation on hematological status, cell-mediated immunity and susceptibility to infections in children living in an environment where iron deficiency, malaria and other infections are frequent. DESIGN: Randomized, double-blind iron supplementation including a placebo group. SETTING: A village in Togo, West Africa. SUBJECTS: Of the 229 6-36-month-old children of both sexes recruited, 197 with hemoglobin concentration >/=80 g/l were included and 163 completed the study. INTERVENTION: Children received daily a placebo (n=79) or a dose of 2-3 mg of elemental iron per kg of body weight (n=84) for 3 months. Hematological, nutritional and immune status were assessed at the beginning and at the end of the supplementation period, and 6 months later. Morbidity was recorded throughout the study. RESULTS: Iron supplementation had a significant and positive effect on iron status of children and no impact on the incidence of infections, especially malaria. Its probable effect on immune status was masked by interference of infections and their treatment, which contributed to improve hematological and immune status in both groups. CONCLUSION: According to the negative consequences of anemia and iron deficiency on global child development, control of iron deficiency by oral iron supplementation in young children has to be conducted, associated with prophylaxis and treatment of malaria and repeated deworming. SPONSORSHIP: Program supported by IRD. European Journal of Clinical Nutrition (2000) 54, 29-35 PMID- 10694770 TI - Age patterns in stunting and anaemia in African schoolchildren: a cross-sectional study in Tanzania. AB - OBJECTIVE: To describe the nutritional status of schoolchildren from a rural area of Tanzania, with a particular emphasis on older adolescents to determine the timing of the growth spurt and differences by sex. DESIGN: A cross-sectional survey using a randomly selected sample. SUBJECTS: Six thousand eight hundred and one children aged 7-18 y randomly selected from those enrolled in standards 2-5 in 59 primary schools in Magu District, Tanzania. RESULTS: Overall, 52.5% of children were stunted and 43.0% were underweight, with significantly more boys stunted and underweight than girls. Z-scores of height-for-age for both boys and girls decreased progressively between 7 and 12 y. After 12 y the height-for-age z scores of girls show a marked upturn, whilst z-scores for boys continue to decrease throughout the school-aged years until 16 y when a slight upturn is observed. Anaemia (Hb<120 g/L) was present in 62.6% of children, with the prevalence decreasing with age. Anaemia improved throughout the school years for boys, but did not for girls. Age, sex and hookworm infection were significant predictors of anaemia. CONCLUSION: Stunting and anaemia are exceptionally common conditions in African schoolchildren. The findings highlight important differences between boys and girls, which are suggestive of compensatory growth at 12 y for girls and at 16 y for boys, although it remains unclear whether boys will catch up in height at older ages. SPONSORSHIP: Funding was provided by the Wellcome Trust. European Journal of Clinical Nutrition (2000) 54, 36-40 PMID- 10694771 TI - Predictors of growth from 1 to 18 months among breast-fed Ghanaian infants. AB - OBJECTIVE: To examine factors associated with the physical growth of breast-fed Ghanaian infants during the first 18 months of life. DESIGN: A community-based longitudinal study. SETTING: The study was carried out in Techiman, a district capital and major food trading center in the Brong Ahafo region of Ghana. SUBJECTS: One-month old infants (n=216) with birth weight >/= 2.5 kg were recruited from Maternal and Child Health Centers. METHOD: From 6 to 12 months, infants were provided with one of four types of nutritionally enhanced complementary foods. Anthropometric assessments were completed monthly from 1 to 12 months and every other month from 12 to 18 months. Information was collected on household characteristics, morbidity from common infections and dietary intakes. Blood samples were collected at 6 and 12 months to assess iron, zinc, riboflavin and vitamin A status. Multiple regression analysis was used to examine factors associated with growth during the age intervals of 1-6, 4-6, 6-12 and 12 18 months as well as size attained at 12 and 18 months. RESULTS: Prevalence of diarrhea and fever were negatively associated with growth during the first year of life. No significant relationship was found between respiratory illness (defined as cough or purulent nasal discharge) and growth. With the exception of dietary zinc intake, dietary variables were generally not significantly associated with growth. Maternal education was positively associated with growth during most of the age intervals. CONCLUSION: These findings suggest that interventions to reduce morbidity and improve the education of girls may benefit children's growth in this population. SPONSORSHIP: Nestle Foundation; Rockefeller Foundation African Dissertation Internship Award; Fulbright Scholarship. European Journal of Clinical Nutrition (2000) 54, 41-49 PMID- 10694772 TI - Arachidonic and docosahexaenoic acids are strongly associated in maternal and neonatal blood. AB - BACKGROUND: The red cell membrane fatty acid composition has frequently been used as an index of essential fatty acid (EFA) nutrition. After birth there is a decline in plasma arachidonic acid (AA) and docosahexaenoic (DHA) acids in babies fed on conventional formula which contains only the parent linoleic and alpha linolenic acids. In human studies, the red cell phosphoglyceride composition appears to be more constant than that of plasma. In infants fed fish oil without AA, the AA proportions fall in the plasma but much less so in the red cells. This result might be considered to mean that there is no need for preformed AA. On the other hand, in a study where the levels of AA fell there was reduction of infant growth. Indeed, where cell membrane composition does change there is often an associated alteration in physiological functions of membranes. We therefore felt it worth investigating the balance between AA and DHA in a physiological situation where plasma levels are known to change, namely in pregnancy. PURPOSE: The aim of the study was to investigate a relationship between blood phosphoglyceride AA and DHA in pregnant women and neonates. SUBJECTS: Health pregnant women from London, England (n=193) and their term babies (n=45); healthy pregnant women from Seoul, South Korea (n=40) and their term babies (n=40); and preterm neonates (n=72) from London. METHOD: Blood samples were taken from British and Korean pregnant women during the third trimester, and from term and preterm babies at birth. These samples were taken for routine monitoring purposes in Korea and were a part of a study on pregnancy outcome for which ethical permission was granted from the East London and The City Health Authority and Lambeth, Southwark and Lewisham Health Authority. Approval was also obtained from the Ethical Committee of the Asan Medical Centre, Seoul, South Korea. RESULTS: AA and DHA correlated in plasma choline phosphoglycerides (CPG) of the British mothers (r=0.52 P<0.0001). The correlation coefficients and significance were much stronger in the red cell CPG and even more so in the term and preterm infant red cell CPGs ( r=0.75, 0.80 and 0.88, respectively). Similarly, AA and DHA correlated in red cell CPGs of the Korean women and their term babies. There was also a significant relationship between the two fatty acids in red cell ethanolamine phosphoglycerides in the mothers and their babies. Both linoleic (LA) and alpha-linolenic acids (ALA) were inversely associated with AA and DHA in some of the phosphoglyceride fractions of the mothers and babies. CONCLUSIONS: Although AA and DHA have different primary dietary origins, there were significant relationships between AA and DHA in the phosphoglycerides of the red cell membrane. This finding seems surprising if the red cell composition is determined by diet. These results suggest a physiological mechanism which attempts to maintain an appropriate balance between AA and DHA. It is plausible that there is an optimum balance between AA and DHA for membrane stability, deformability, enzyme and receptor function. SPONSORSHIP: The British Diabetic Association, March of Dimes Birth Defects Foundation and The Christopher H.R. Reeves Charitable Trust. European Journal of Clinical Nutrition (2000) 54, 50-56 PMID- 10694773 TI - Iodine content in drinking water and other beverages in Denmark. AB - OBJECTIVE: To investigate the variation in iodine content in drinking water in Denmark and to determine the difference in iodine content between organic and non organic milk. Further, to analyse the iodine content in other beverages. DESIGN AND SETTING: Tap water samples were collected from 41 evenly distributed localities in Denmark. Organic and non-organic milk was collected at the same time (twice summer and twice winter). Soft drinks, beers and juice were collected from different Danish producers and wine from different countries. All samples were analysed for iodine using inductively coupled mass spectrometry. RESULTS: Iodine in tap water varied from 2.1 to 30.2 microg/l; the iodine content was in general highest in the eastern part of Denmark and lowest in the western part of Denmark. Organic milk was found to have a lower iodine content than non-organic milk. CONCLUSIONS: Large geographical (and seasonal) variations in iodine concentrations were found in different beverages supplying an appreciable part of the iodine in the Danish diet. This knowledge is important when calculating the iodine intake from dietary intake studies. SPONSORSHIP: The 1991 Farmacy Foundation and Danish Veterinary and Food Administration. European Journal of Clinical Nutrition (2000) 54, 57-60 PMID- 10694774 TI - Comparison of an oleic acid enriched-diet vs NCEP-I diet on LDL susceptibility to oxidative modifications. AB - OBJECTIVE: The objective of this trial was to compare the effect on the susceptibility of plasma Low Density Lipoprotein (LDL) to oxidative modifications of consumption of two oleic rich diets, prepared with two different plant oils, virgin olive oil (OL)1 and refined high monounsaturated fatty acids (MUFA sunflower oil (SU)), with the susceptibility of plasma LDL to oxidation after an National Cholesterol Education Program step 1 (NCEP-I) phase diet. DESIGN: A randomized crossover design. SUBJECTS AND INTERVENTIONS: Twenty-two healthy normolipidemic young males consumed an NCEP-I diet for a 4-week period. Subjects were then assigned to two diets each of 4-weeks duration. Group one was placed on an olive oil enriched diet (40% fat, 22% MUFA) followed by a 4-week period of a MUFA diet enriched in sunflower oil (40% fat, 22% MUFA). In group two, the order of the diets was reversed. RESULTS: Both MUFA diets induced a decrease in saturated (14:0, 16:0, and 18:0) and an increase in monounsaturated and polyunsaturated n-6 (18:2, 20:3, and 20:5) plasma LDL-phospholipid fatty acids, compared to the NCEP-I diet (P<0.01). No significant differences in lag times were observed between the olive oil and the NCEP-I diet periods. However there was a greater inhibition time (P<0.001) when subjects consumed the MUFA rich sunflower oil diet compared to the NCEP-I diet. These differences were probably related to the relative enrichment of plasma LDL particles in alpha-tocopherol due to the high vitamin E content of the MUFA-rich sunflower oil. Indeed, the alpha-tocopherol content was positively correlated with lag time (r=0.338; P<0.008). CONCLUSION: Our findings suggest that changes in plasma LDL alpha tocopherol content with practical solid-food diets can decrease its susceptibility to oxidation. SPONSORSHIP: This work has been supported by grants from the Investigaciones de la Seguridad Social (FIS 92/0182, to Francisco Perez Jimenez); and from Koype Co, Andujar, Jaen, Spain. European Journal of Clinical Nutrition (2000) 54, 61-67 PMID- 10694775 TI - Vitamin D status and parathyroid hormone concentrations in Chinese women and men from north-east of the People's Republic of China. AB - STUDY OBJECTIVE: To evaluate vitamin D status of young and old women and men living in Shenyang, north-east People's Republic of China in early spring and to explore the relationship between vitamin D metabolites and parathyroid hormone (PTH) in this population. DESIGN: Cross-sectional study. SETTING: Shenyang, north east China. SUBJECTS: 194 healthy volunteers: 48 young women and 48 young men aged 25-35 y, and 48 old women and 50 old men aged 65-75 y. RESULTS: Fasting blood samples were used to measure plasma 25-hydroxyvitamin D (25(OH)D), 1,25 dihydroxyvitamin D (1,25(OH)2D) and PTH using radioimmunoassays. The proportion of subjects who could be regarded as vitamin D deficient (<25 nmol/l) was 48%, 29%, 15% and 13% for old men, young men, old women and young women, respectively. There was no association between plasma 1,25(OH)2D and 25(OH)D concentrations. PTH concentrations were elevated in both old women and men compared with young subjects (P<0.01). A negative association between PTH and 25(OH)D was only found in old women (P<0. 001), but not in old men, nor in young subjects. CONCLUSIONS: Vitamin D status was poor in this population in early spring, especially in men. There was no clear evidence to show that the secretion of PTH and the conversion of 1,25(OH)2D were affected by the low 25(OH)D concentration. SPONSORSHIP: Partly supported by the Sandoz Foundation for Gerontological Research and the Nestle Foundation, Switzerland, and Medical Research Council, UK. European Journal of Clinical Nutrition (2000) 54, 68-72 PMID- 10694776 TI - Ten-year trends in vitamin and mineral intake from fortified food in German children and adolescents. AB - OBJECTIVE: We investigated time trends in consumption patterns, and energy and nutrient intakes (protein, fat, carbohydrates, added sugars, vitamins A, E, C, B1, B2 and B6, niacin, folate, calcium and iron) from fortified food in children and adolescents between 1987 and 1996 in Germany. DESIGN: Mixed longitudinal survey (DONALD study) with 3 d weighed dietary records (n=2062 from 594 subjects), one subject per family per year chosen by random. SETTING: Dortmund (Western Germany) district cohort. SUBJECTS: 285 males, 309 females; mean age 6 y (2-13 y). RESULTS: Almost all children and adolescents consumed fortified food irrespective of the year studied. With the exception of vitamin E, significant time trends in the proportions of nutrient intakes from fortification were observed. The fortification of food with vitamins A, C, B1, B2 and B6 and niacin raised the already adequate intakes from non-fortified food (100% to 150% of reference intake values) by 20-50%. The fortification of food with vitamin E and folate raised the low intakes from non-fortified food (about 50% of reference intake values) to about 80% (folate) and 100% (vitamin E) of the references. Fortification of food with calcium and iron was not significant (<10%), but while total intake of calcium was adequate, total intake of iron remained critical. CONCLUSIONS: Since the nutrient intake of the population of children and adolescents studied is adequate with respect to vitamins A, C, B1, B2 and B6, niacin and calcium, fortification seems inefficient, while fortification of food with vitamin E and folate, but not iron, improves an inadequate intake. SPONSORSHIP: The DONALD study is supported by the German Federal Ministry of Health and the North-Rhine-Westphalian Ministry of Science and Research. European Journal of Clinical Nutrition (2000) 54, 81-86 PMID- 10694777 TI - A single dose of tea with or without milk increases plasma antioxidant activity in humans. AB - OBJECTIVE: To investigate the effect of black and green tea consumption, with and without milk, on the plasma antioxidant activity in humans. DESIGN: In a complete cross-over design, 21 healthy volunteers (10 male, 11 female) received a single dose of black tea, green tea (2 g tea solids in 300 ml water) or water with or without milk. Blood samples were obtained at baseline and at several time points up to 2 h post-tea drinking. Plasma was analysed for total catechins and antioxidant activity, using the ferric reducing ability of plasma (FRAP) assay. RESULTS: Consumption of black tea resulted in a significant increase in plasma antioxidant activity reaching maximal levels at about 60 min. A larger increase was observed after consumption of green tea. As anticipated from the higher catechin concentration in green tea, the rise in plasma total catechins was significantly higher after consumption of green tea when compared to black tea. Addition of milk to black or green tea did not affect the observed increases in plasma antioxidant activity. CONCLUSIONS: Consumption of a single dose of black or green tea induces a significant rise in plasma antioxidant activity in vivo. Addition of milk to tea does not abolish this increase. Whether the observed increases in plasma antioxidant activity after a single dose of tea prevent in vivo oxidative damage remains to be established. European Journal of Clinical Nutrition (2000) 54, 87-92 PMID- 10694778 TI - Dietary habits among patients with atopic dermatitis. AB - OBJECTIVE: To evaluate the dietary habits among adult patients with moderate to severe atopic dermatitis and relate intake to clinical symptoms. DESIGN: Data were obtained from a clinical trial. SETTING: Five departments of dermatology at Norwegian University hospitals. SUBJECTS: Outpatients, 46 men (median age 27 y) and 92 women (median age 28 y). METHOD: A quantitative food frequency questionnaire was filled in before attending the clinical trial. The results were compared to the diet of age- and sex-matched reference groups. RESULTS: Male patients had higher content of refined sugar in their diet than reference men (P=0.014). Among female patients, the intake of saturated fatty acids was higher (P=0.049), whereas the intake of very long-chain n-3 fatty acids was lower (eicosapentaenoic acid, P=0.032, docosahexaenoic acid, P=0.017) than in the reference group. In both genders, more patients than reference subjects had vitamin D intake below recommended level. Furthermore, the female patients had significantly lower intake of fruit compared to the reference group (P=0.002). No correlation was found between nutrient intake of the patients and their clinical scores. CONCLUSIONS: The patients's diet were fairly similar to the diet of reference groups. The intake of vitamin D and very long-chain n-3 fatty acids was low, especially among female patients. Furthermore, we could not detect any association between dietary habits and clinical status. European Journal of Clinical Nutrition (2000) 54, 93-97 PMID- 10694779 TI - Toenail selenium and breast cancer-a case-control study in Finland. AB - OBJECTIVE: Low levels of selenium have been associated with a higher risk of cardiovascular diseases and cancer in humans. Since 1984, selenium supplementation through fertilizers has been employed in Finland to increase the very low concentration of selenium in the nation's food supply. As a result, the selenium concentration of Finnish foods became one of the highest in Europe. A decade after selenium supplementation began, the association between toenail selenium and the risk of breast cancer was examined. DESIGN: Case-control study. SETTING: Eastern Finland. SUBJECTS: 289 pre- and postmenopausal breast cancer cases and 433 community controls. The diagnosis was unknown at the time the toenail samples were collected. RESULTS: The mean toenail selenium concentration was 0.80 mg/kg in premenopausal cases and 0.84 mg/kg in premenopausal controls: and 0. 77 mg/kg in postmenopausal cases and 0.80 mg/kg in postmenopausal controls. The odds ratio (OR) comparing the highest with the lowest quintiles of toenail selenium concentration was 1.1 (95% CI 0.4-3.2) in premenopausal women and 0.7 (95% CI 0.3-1.5) in postmenopausal women. The intake of retinol, beta carotene, vitamin E and vitamin C did not change the association between toenail selenium and breast cancer. CONCLUSIONS: A decade after selenium supplementation, selenium seems not to be an important factor in the etiology of breast cancer, neither in premenopausal nor postmenopausal women. SPONSORSHIP: This work was supported by the EVO funds from the Kuopio University Hospital and by research grants from the Academy of Finland, Yrjo Jahnsson Foundation and Juho Vainio Foundation. European Journal of Clinical Nutrition (2000) 54, 98-103 PMID- 10694780 TI - The effect of dietary trans alpha-linolenic acid on plasma lipids and platelet fatty acid composition: the TransLinE study. AB - OBJECTIVE: To collect (i) baseline data and (ii) execute a large multicentre study examining the effect of trans alpha-linolenic acid on its incorporation into plasma lipids and on risk factors for coronary heart disease. DESIGN: Male volunteers were recruited and the habitual diet assessed by a 4-d weighed record. Fatty acid composition of plasma and platelet lipids were determined by gas chromatography at baseline. After a 6 week run-in period on a trans 'free' diet, male volunteers were randomised to consume 0.6 % of energy trans alpha-linolenic acid or to continue with a diet 'low' in trans alpha-linolenic acid for 6 weeks. SETTING: Three European university research departments supported by the research and development departments of the food industry. SUBJECTS: Male volunteers (88) recruited by local advertisement. METHODS: Replacement of 30 % of the fat of the habitual diet by margarine, oil and foods. Rapeseed oil was deodorised especially to produce the trans 'free' and 'high' trans foods for this study. The incorporation and conversion of trans alpha-linolenic acid into plasma lipids and platelets was assessed by gas chromatography and dietary compliance was verified by 4-d weighed record. RESULTS: Less trans alpha-linolenic acid isomers are incorporated into human plasma lipids in French volunteers than in Dutch or Scottish volunteers consuming their habitual diets. Trans 'free' alpha-linolenic acid-rich oil can be produced by careful deodorization during refining. The 'high' trans diet provided 1410+/-42 mg/d trans isomers of alpha-linolenic acid, whilst the 'low' trans group consumed 60+/-75 mg/d. The change in plasma lipid and platelet fatty acid composition documented that trans linolenic isomers are incorporated and converted to a trans isomer of eicosapentaenoic acid. Only the 15-trans alpha-linolenic acid is incorporated into plasma cholesteryl esters. The group consuming low trans diet had a slightly higher intake of fat, especially saturated and monounsaturated fat. CONCLUSIONS: Trans 'free' rapeseed oil, rich in alpha-linolenic acid, can be produced by careful deodorization. Dietary records show good compliance. Dietary trans isomers of alpha-linolenic acid are incorporated in plasma lipids and converted to long-chain polyunsaturated fatty acids. Their effects on risk factors for coronary heart disease and their metabolism will be reported elsewhere. SPONSORSHIP: European Commission (FAIR 95 0594 grant). European Journal of Clinical Nutrition (2000) 54, 104-113 PMID- 10694781 TI - Infant-feeding patterns are related to blood cholesterol concentration in prepubertal children aged 5-11 y: the Fleurbaix-Laventie Ville Sante study. AB - OBJECTIVE: Several studies, mainly in animals, but also in humans, have shown that diet in infancy is associated with differences in blood cholesterol concentrations later in life. The objective was to examine this relationship in children aged 5-11 y after taking into account their current diet and parental hypercholesterolemia. SETTING AND SUBJECTS: 251 prepubertal boys and 223 prepubertal girls enrolled in the schools in two little towns in northern France. DESIGN AND METHODS: Cross-sectional evaluation including measurements of cholesterol concentrations on capillary blood and a single weekday food intake record. Infant feeding patterns were obtained by questionnaire given to the mothers. RESULTS: 50% of the children had been breast-fed for a median duration of less than 2 months. Cow's milk was introduced in the diet as whole milk for 33% of the children. After adjustment for age, height, and sibship, capillary cholesterol concentration was lower in boys who had been breast fed (geometric mean: 4.4, 95% confidence interval of the mean: 4.2-4.6 mmol/L) than in those fed with formula (4.7, 4.5-4.8 mmol/L, P<0.03). In girls, breastfeeding had no significant effect on blood cholesterol concentration, which was associated with the type of cow's milk given in infancy: whole milk: 4.9 mmol/L (4.7-5. 2); totally or partially skimmed milk: 4.5 mmol/L (4.2-4.6), P<0.008. The current saturated fat and cholesterol intakes and parental hypercyholesterolemia were associated with current blood cholesterol concentration in children, but did not modify its relationship with infant feeding patterns. CONCLUSION: Results of the present study suggest that diet in infancy may have longstanding effect on lipid metabolism. SPONSORSHIP: The study was supported by funds from Eridania Beghin Say, Groupe Fournier, Lesieur and Nestle France, Roche Diagnostic and of the MGEN (Mutuelle Generale de l'Education Nationale, contract INSERM-MGEN #9158) and a grant from the Association de Langue Francaise pour l'Etude du Diabete et du Metabolisme (ALFEDIAM). European Journal of Clinical Nutrition (2000) 54, 114-119 PMID- 10694782 TI - Digestibility of cocoa butter from chocolate in humans: a comparison with corn oil. AB - OBJECTIVE: To compare, in humans, the digestibility of moderate amounts of cocoa butter (30.7 g/d) consumed in the form of chocolate as part of a normal western diet with that of a well-absorbed fat (corn oil); and hence determine whether, by virtue of its apparent low absorption, cocoa butter can be considered to be a low calorie fat. DESIGN: Randomised, two-period crossover metabolic study, conducted under free-living conditions, but with strict control over food intake. SETTING: Metabolic Unit, Nestle Research Center Lausanne. SUBJECTS: Twelve healthy men were selected from volunteers at the Nestle Research Center and all subjects completed the study. INTERVENTION: Two treatment periods of two weeks each: cocoa butter and control periods, with strict dietary control separated by a two week wash out period. RESULTS: No differences (P>0.05) were observed in faecal weight (wet or dry), faecal fat nor in defecation frequency between treatments (cocoa butter and corn oil). Cocoa butter at a dose of 30.7 g/d in the form of black chocolate, consumed between two meals, was found to have a similar digestibility to that of corn oil (99 % of corn oil digestibility). CONCLUSION: Cocoa butter, consumed as back chocolate within a normal mixed diet, has a high digestibility, similar to that of corn oil, and a digestible energy value of 37 kJ/g in man. Thus, cocoa butter cannot be considered to be a low-calorie fat. SPONSORSHIP: Nestec Ltd, Switzerland. European Journal of Clinical Nutrition (2000) 54, 120 125 PMID- 10694783 TI - Association between trans fatty acid intake and cardiovascular risk factors in Europe: the TRANSFAIR study. AB - BACKGROUND: High intakes of trans fatty acids (TFA) have been found to exert an undesirable effect on serum lipid profiles, and thus may increase the risk for cardiovascular disease. OBJECTIVES: Investigation of the association between TFA intake and serum lipids. DESIGN: Cross-sectional study in eight European countries (Finland, France, Greece, Iceland, The Netherlands, Portugal, Spain, Sweden) among 327 men and 299 women (50-65 y). Using a dietary history method, food consumption was assessed and TFA intake was calculated with recent figures on TFA levels of foods, collected in the TRANSFAIR study. RESULTS: Mean (+/-s.d.) TFA intake was 2.40+/-1.53 g/day for men and 1.98+/-1.49 g/day for women (0.87+/ 0.48% and 0. 95+/-0.55% of energy, respectively), with the highest consumption in Iceland and the lowest in the Mediterranean countries. No associations were found between total TFA intake and LDL, HDL or LDL/HDL ratio after adjustment for cardiovascular risk factors. Additional adjustment for other fatty acid clusters resulted in a significant inverse trend between total TFA intake and total cholesterol (Ptrend<0.03). The most abundantly occurring TFA isomer, C18:1 t, contributed substantially to this inverse association. The TFA isomers C14:1 t9, C16:1 t9 and C22:1 t were not associated or were positively associated with LDL or total cholesterol. CONCLUSIONS: From this study we conclude that at the current European intake levels of trans fatty acids they are not associated with an unfavourable serum lipid profile. SPONSORSHIP: Unilever Research Laboratorium, the Dutch Dairy Foundation on Nutrition and Health, Cargill BV, the Institute of Food Research Norwich Laboratory, the Nutrition Branch of the Ministry of Agriculture, Fisheries and Food, the International Fishmeal and Oil Manufacturers' Association, Kraft Foods, NV Vandemoortele Coordination Center, Danone Group, McDonalds Deutschland Inc, Danish Veterinary and Food Administration, Valio Ltd, Raisio Group. European Journal of Clinical Nutrition (2000) 54, 126-135 PMID- 10694784 TI - Daily consumption of (red) meat or meat products in Switzerland: results of the 1992/93 Swiss Health Survey. AB - OBJECTIVE: The present study aimed to examine the frequency of daily meat and meat product consumption and the preference for red meat in Switzerland. DESIGN: Cross-sectional Study. SETTINGS AND SUBJECTS: Data were taken from the 1992/1993 Swiss Health Survey, which collected data on a random sample of persons aged 15 and over, living in Switzerland. The survey, which had a response rate of 71%, included 7930 male and 7358 female respondents. Bivariate analyses and multivariate logistic regressions controlling for sociodemographic and lifestyle factors were performed. RESULTS: Daily consumption of meat or meat products (25%) and more frequent consumption of red than white meat (26% of meat eaters) were prevalent in Switzerland. Men, middle-aged persons, participants with a low level of education, persons living in the German or Francophone regions of Switzerland, those with Swiss nationality, smokers, overweight and obese people, those with daily alcohol consumption and physically inactive persons were found to consume daily meat or meat products more frequently. A preference for red meat rather than white meat was more often observed in men, young people, persons living in the German or Francophone regions of Switzerland, smokers and participants who consumed alcohol at least once daily. CONCLUSIONS: The analysis of data from the 1992/1993 Swiss Health Survey shows that in specific subgroups of the Swiss population meat and meat product consumption is still more frequent than recommended, but crude comparisons with older and more recent studies indicate a decrease in meat consumption. The observed clustering of daily meat consumption with other risk factors underscores the necessity to include dietary recommendations in health programs addressing other unhealthy lifestyles. European Journal of Clinical Nutrition (2000) 54, 136-142 PMID- 10694785 TI - Prediction of dietary flavonol consumption from fasting plasma concentration or urinary excretion. AB - OBJECTIVES: to predict flavonols content of the habitual diets of free-living subjects from urine and plasma concentrations of flavonols. DESIGN: Ten type 2 diabetic patients (five male, five female), mean age 60 (s.e.m. 7) y and BMI 30.2 (s.e.m. 3.5) kg/m2 were treated in a random crossover design for a 2 week period on either a low flavonoid diet or on the same diet supplemented at one of two high flavonols levels (total 77.3 or 110.4 mg/day) provided by supplements of 1500 ml tea daily and 400 g fried white onion in olive oil with and without tomato ketchup and herbs. SETTING: Glasgow Royal Infirmary, University of Glasgow, Scotland. MAIN OUTCOME MEASURES: Fasting plasma concentration, urine concentration and 24 h excretion of quercetin, isorhamnetin, kaempferol and myricetin. RESULTS: Plasma flavonol concentration (r=0.750, P=0.001), 24 h urine concentration (r=0.847, P=0.001) and 24 h urine excretion (r=0.728, P=<0.001) were all highly significantly related to dietary intake and gave similar estimates of intakes. Fasting plasma flavonols concentrations on habitual diets ranged from 0 to 43.7 ng/ml mean. Regression equations were constricted: total flavonols intake r=0.74, P<0.001 and quercetin intake r=0.744, P<0. 001. From these equations, flavonol intakes from habitual diets were estimated at 17-50, mean 35 mg/day. Of this, 91% was from quercetin. CONCLUSIONS: Dietary flavonols are absorbed and appear in plasma and urine as potential biomarkers in concentrations related quantitatively to intake. Estimation of dietary intake from plasma or urine concentrations appears possible. SPONSORSHIP: Rank Prize Funds and Rank Foundation of the Department of Human Nutrition; Ministry of Health and Medical Education, IR Iran. European Journal of Clinical Nutrition (2000) 54, 143-149 PMID- 10694786 TI - The habitual diet in rural and urban Cameroon. AB - OBJECTIVE: To evaluate the habitual diet of a rural and urban population in Cameroon, Central Africa. SETTING: An urban area-Cite Verte Housing District, Yaounde (1058 subjects); and a rural area-three villages in Evodoula, Cameroon (746 subjects). SUBJECTS: Cameroonian men and women of African origin (1058 urban, and 746 rural), aged 24-74 y. METHODS: The habitual diet was estimated with an interviewer-administered food frequency questionnaire. MAIN OUTCOME MEASURES: Macro- and micronutrient intake. RESULTS: The intake of energy, fat and alcohol was higher in rural men and women than in urban subjects. In rural women, the intake of carbohydrates and protein was also higher. The intakes of fibre, iron, carotene, zinc, potassium, and of the vitamins C, D and E were all higher in rural men and women than in their urban counterparts. The intake of retinol was lower in rural subjects than in urban subjects. Eight of the 10 foods eaten in the highest amount and contributing most to energy intake differed between the rural and urban population. CONCLUSION: The habitual diet in rural Cameroon contains more fat and alcohol than the diet in urban Cameroon. The high physical activity in the rural area may explain the lower levels of the cardiovascular risk factors in this area compared to those of the urban dwellers. SPONSORSHIP: This work was supported by a grant from the European Union (contract no. TS3*CT92 0142) and by the Conseil Regional d'Ile de France and INSERM. European Journal of Clinical Nutrition (2000) 54, 150-154 PMID- 10694787 TI - Histamine plasma levels and elimination diet in chronic idiopathic urticaria. AB - OBJECTIVE: The aim of this study was to evaluate the effects of an oligoantigenic and histamine-free diet on patients affected with chronic idiopathic urticaria (CIU). DESIGN: Ten patients with chronic idiopathic urticaria were prescribed an oligoantigenic and histamine-free diet for 21 days, followed by serial and controlled reintroduction of foods during a further 70 days. Modification in clinical illness as well as histamine plasma levels, post-heparin plasma diamine oxidase (DAO) and intestinal permeability were evaluated. RESULTS: The oligoantigenic and histamine-free diet induced a significant improvement of symptoms (P<0.05). Moreover, CIU patients on free diet showed higher histamine plasma levels (P<0. 05 vs post-diet and vs controls) that fell to control levels during the oligoantigenic and histamine-free diet. Post-heparin plasma diamine oxidase values were slightly reduced and were unchanged during the diet as well as intestinal permeability, which was always normal in all patients. CONCLUSIONS: These data suggest that histamine plays a major role in chronic idiopathic urticaria. The finding of normal intestinal permeability suggests that a morphological damage of intestinal mucosa should be excluded in these patients. However, the presence of low levels of post-heparin plasma diamine oxidase may indicate a subclinical impairment of small bowel enterocyte function that could induce a higher sensitivity to histamine-rich or histamine-producing food. European Journal of Clinical Nutrition (2000) 54, 155-158 PMID- 10694788 TI - Education and nutrient intake in Dutch elderly people. The Rotterdam Study. AB - OBJECTIVE: Unfavourable dietary habits might explain a part of the increased cardiovascular morbidity and mortality among the lower socioeconomic groups. The aim of the study was to describe differences in dietary intake in older subjects by socioeconomic status, as indicated by educational level. DESIGN: A cross sectional analysis of socioeconomic status in relation to dietary intake. SETTING: The Rotterdam Study. SUBJECTS: 2213 men and 3193 women, aged 55 y and over living between 1990 and 1993 in a district of Rotterdam, The Netherlands. METHODS: Dietary data were assessed with a semiquantitative food frequency questionnaire, containing 170 food items in 13 food groups. RESULTS: In general, the dietary differences between socioeconomic groups were small. Lower educated subjects had a higher intake of almost all macronutrients compared with higher educated subjects. The total energy intake of men/women with the lowest educational level differed from those with the highest education in the following respect: 9.60/7.54 vs 8.94/7.17 MJ/day. Furthermore, fat composition was more adverse in the lower educated strata; in lower educated subjects, relatively more energy was derived from saturated fat (14.5/14.6 vs 13.8/13.8 energy%), the ratio of polyunsaturated saturated fat was lower (for men: 0.50 vs 0. 55) and the intake of cholesterol higher (271/220 vs 240/204 mg/day). These differences could be explained by a higher intake of visible fat (46/37 vs 44/34 g/day) and more meat consumption (130/100 vs 116/86 g/day). In addition, the composition of these products differed: the higher educated used relatively more lean meat and low-fat milk products. Furthermore, the intake of fibre was lower among the lower educated (1.88/2.17 vs 2.03/2.29 g/MJ). Among lower educated groups there were more abstainers (15.5/31.5 vs 12.3/26.9%) and the type of alcoholic beverages also differed between the groups. Intake of antioxidant vitamins from food alone did not differ between educational groups. CONCLUSIONS: In Dutch elderly people, there are socioeconomic differences in dietary intake. Although these differences are small, these findings support the role of diet in the explanation of socioeconomic inequalities in cardiovascular health. SPONSORSHIP: Erasmus Centre for Research on Aging, Erasmus University Rotterdam. European Journal of Clinical Nutrition (2000) 54, 159-165 PMID- 10694789 TI - The influence of survey duration on estimates of food intakes and its relevance for public health nutrition and food safety issues. AB - OBJECTIVE: To examine the influence of food consumption survey duration on estimates of percentage consumers, mean total population intakes and intakes among consumers only and to consider its relevance for public health nutrition and food safety issues. DESIGN: Prospective food consumption survey. SETTING: A multicentre study in five centres in the European Union-Dublin, Ghent, Helsinki, Potsdam and Rome. SUBJECTS: Teenage subjects were recruited through schools; 948 (80%) out of 1180 subjects completed the survey. INTERVENTIONS: 14-day food diaries were used to collect the food consumption data. RESULTS: For mean total population intakes, 53% of the foods had slopes significantly different to 0 (P<0.05). In practical terms (g/day), these differences were small, with 41% of foods having differences of or =160 mm Hg and diastolic blood pressure (DBP) < or =90 mm Hg) or normotension (n = 124, SBP <140 mm Hg and DBP <90 mm Hg with no family history of hypertension). To collect demographic data, a structured questionnaire was used. DNA was extracted using standard techniques from 211 subjects (age range 61-89, mean age 73 +/- 6.6 years, male: female ratio 1.1:1). Genotypes (gly/gly, trp/gly and trp/trp) were assigned in triplicate by polymerase chain reaction (PCR) followed by electrophoresis, using a laser scanning electrophoresis unit. The validity of the method was confirmed by sequencing. Frequencies of allele distribution in ISH or control groups were determined by Chi-square tests and a stepwise logistic regression model, which controlled for potential confounders, was used to examine any independent association between alpha-adducin genotypes or alleles with ISH and normotensive controls. Mean BP (+/- s.d.) was: 170/79.8 +/- 14.9/8.3 mm Hg and 122.1/ 73.4 +/- 8. 8/7.6 mm Hg in the ISH and normotension groups respectively. The unadjusted allele and genotypes frequencies were not significantly different in the ISH patients groups compared with normotensive controls (chi2 = 1.59, P = 0.45 and chi2 = 1.23, P = 0.28 respectively). In this elderly cohort, after adjustment for potential confounders, no statistically significant association was found between alpha adducin genotype and SBP (P = 0.65 for homozygotes, P = 0.59, for heterozygotes), DBP (P = 0.49 homozygotes, for heterozygotes P = 0.45) pulse pressure (P = 0.87 homozygotes, for heterozygotes P = 0.95) diagnosis of ISH (P = 0.72 for homozygotes, P = 0.68 for heterozygotes). However age and renal disease predicted the diagnosis of ISH (P = 0.001, P = 0.459, respectively), a large pulse pressure (P < 0.0001, P = 0.033, respectively) and a higher SBP (P < 0.0001, P = 0.025, respectively) in this large cohort of elderly Australian Caucasian volunteers. Journal of Human Hypertension (2000) 14, 199-203. PMID- 10694836 TI - Bisoprolol and nifedipine retard in elderly hypertensive patients: effect on quality of life. AB - Subjects over the age 60 with sustained sitting diastolic pressures of 95-115 mm Hg were randomised to a regime based on bisoprolol (n = 368) or nifedipine retard (n = 379) for 24 weeks. The goal diastolic pressure was < or =90 mm Hg and to achieve this, double-blind medication could be doubled (5/10 mg bisoprolol, 40/80 mg nifedipine retard) or hydrochlorothiazide 25 mg (unblinded) could be added to the higher dose. In an intention-to-treat analysis, 309 subjects in both the bisoprolol and nifedipine retard treated group provided at least a baseline and a second quality of life assessment (82%). An excess of symptoms was observed in the nifedipine group for oedema of the legs, nocturia, constipation, racing heart and heart thumping. Fewer patients reported wheeze in the nifedipine group. For quality of life, there were no statistically significant differences between the two groups after 8 weeks. However, when analysing the results of the last available assessment (usually at 24 weeks) there were significant (P < 0.05) improvements in tension/anxiety, anger/ hostility, vigour/activity, and confusion/bewilderment, assessed by the Profile of Mood States (POMS) in patients receiving bisoprolol in comparison to those receiving nifedipine retard. The Sickness Impact Profile and objective tests of cognitive function did not differ statistically between the two groups. Quality of life was maintained at a good level on both treatments with advantages for bisoprolol in certain areas. Journal of Human Hypertension (2000) 14, 205-212. PMID- 10694837 TI - The association of RBC sodium-lithium countertransport (Vmax) with left ventricular mass in African American women. AB - In Caucasian hypertensives and diabetics, increased RBC sodium-lithium countertransporter activity (SLC) is a marker for end-organ complications of vascular disease. A subgroup of African Americans with high Vmax for SLC show strong correlations with dyslipidaemia, insulin resistance, microalbuminuria and higher blood pressure. The purpose of our study was to determine if Vmax in premenopausal African American women correlates with left ventricular mass (LVM) before the onset of clinically diagnosed hypertension. Non-diabetic African American women (n = 35, mean age 31 years) were evaluated for cardiovascular disease risk factors, including anthropometric and blood pressure measurements, oral glucose tolerance test (OGTT), and euglycaemic hyperinsulinaemic clamp for insulin sensitivity. Fasting blood specimens were assayed for SLC activity (Vmax) and lipids. Cardiac structure was determined by 2-D echocardiography. LVM was calculated by the cube root formula and adjusted for height (LVM index). Vmax correlated significantly with average systolic blood pressure (r = 0.45, P = 0.007), diastolic blood pressure (r = 0.48, P = 0.004), mean blood pressure (r = 0.48, P = 0.003) and LVM index (r = 0.40, P = 0.02). Vmax was also associated with fasting insulin (r = 0.39, P = 0.01), the sum of insulin (r = 0.52, P = 0.002), and insulin sensitivity adjusted for fat-free mass (r = -0.55, P = 0.001). There was no statistically significant relationship between Vmax and body mass or lipids. Vmax for SLC correlates with cardiac structure in premenopausal African American women. Vmax is also associated with insulin sensitivity and insulin resistance in this non-diabetic sample. SLC activity may be useful in identifying a subgroup of young African American women with left ventricular hypertrophy and insulin resistance before the onset of clinically diagnosed hypertension and diabetes. Journal of Human Hypertension (2000) 14, 213-219. PMID- 10694838 TI - High life. A history of high-altitude physiology and medicine PMID- 10694839 TI - Chemokines and chemokine receptors in the pathogenesis of multiple sclerosis. AB - In recent years we have seen growing evidence for the role of chemokines in the pathogenesis of several infectious and non-infectious inflammatory CNS disease states, including Multiple Sclerosis (MS) and its animal model, experimental allergic encephalomyelitis (EAE). An increase in proinflammatory chemokines has been associated with demyelinating lesions and clinical neurological dysfunction in patients with MS; these chemokines could be potential targets for MS therapy. Besides a clearly defined role in mediating leukocyte migration, these and other chemokines may act as immunoregulatory molecules in the driving to Th1/Th2 responses, switch of cytokine profiles, and the induction of tolerance. Since chemokine receptors have now been identified on macrophages, microglia, astrocytes, and endothelial cells as well as neurons in the CNS, chemokine/receptor interactions may mediate functional responses in a variety of CNS cell types during the course of inflammatory disease states. Therefore, clarification of the roles of chemokines and their receptors in the pathogenesis of EAE and MS will be useful in establishing immunotherapeutic strategies for these neurological autoimmune disorders. PMID- 10694840 TI - Evaluation of the apo-1/Fas promoter mva I polymorphism in multiple sclerosis. AB - The pathogenesis of multiple sclerosis is under strong genetic control involving several or more genes each of modest effect. Whilst the mechanisms underlying the pathogenesis of MS remain unknown, it has been hypothesised that either decreased apoptosis of autoreactive T cells in the CNS, or increased apoptosis of oligodendrocytes may play an important role. The Apo-1/Fas antigen (CD95), the gene for which is located in a chromosomal region showing linkage in MS genome screens, is a critical inducer of apoptosis and studies have shown aberrant expression of this molecule in MS, correlating with a decrease in T cell apoptosis or increase in CNS tissue damage. This study investigated an Mva I polymorphism in the Apo-1/Fas promoter region in a group of 124 Australian patients with relapsing-remitting MS and in 183 normal controls. Whilst there were increases in the Mva I*2 allele in MS individuals overall (59% vs 52%, P not corrected=0.08), and in HLA-DRB1*1501 negative MS patients (62% vs 55%), these were not significantly different from controls. Interactions were investigated between the Mva I alleles and T cell receptor beta chain variable region (TCRBV) germline polymorphisms, with a trend in MS individuals towards a decrease of the Mva I*1 allele when combined with the TCRBV3S1*2 allele (Relative Risk=0.25, P=0.067), and with the TCRBV8S1*1 allele (Relative Risk=0.44, P=0.12). Overall, the findings of this study indicate a possible effect of the Apo-1/Fas promoter Mva I polymorphism in MS susceptibility, which needs to be confirmed in further studies. Multiple Sclerosis (2000) 6 14 - 18 PMID- 10694841 TI - Interferon gamma production in blood lymphocytes correlates with disability score in multiple sclerosis patients. AB - The proinflammatory cytokine interferon gamma (IFG) is elevated in body fluids of multiple sclerosis patients but its variation range is broad. The reason for this wide scatter of IFG production is not yet known. We looked for the relation between clinical parameters such as disability, exacerbation frequency, disease duration, course of the disease and IFG producing blood lymphocytes. Forty-one consecutive, clinically stable multiple sclerosis patients with primary relapsing course of the disease and without immunomodulatory or immunosuppressive treatment in the last 3 months were investigated for IFG in blood lymphocytes by flow cytometry. A significant positive correlation between IFG production and disability (r = 0.45, P<0.01, Spearman's rho coefficient) was found. Pathophysiological implications and therapeutical relevance of this unexpected finding are discussed. PMID- 10694842 TI - Positive association between blood brain barrier disruption and osmotically induced demyelination. AB - Rapid correction of chronic hyponatremia can cause osmotic brain demyelination in animals and humans. Why demyelination develops is unknown, but blood brain barrier disruption might expose oligodendrocytes to substances normally excluded from the brain. To test this hypothesis, chronic hyponatremia was induced and corrected using a new, reproducible rat model for producing osmotic brain demyelination. Blood brain barrier integrity was assessed by NMR imaging at either 3, 16 or 24 h during the first day of correction. Demyelination was determined histopathologically 5 - 6 days later. Of 96 rats studied, demyelination developed 5 - 6 days later in 37 rats, 89% of whom showed barrier disruption. In the 59 rats who did not develop demyelination, 45 (76%) had no barrier disruption. Thus, blood-brain barrier disruption during the first 24 h of correction was associated with a 70% risk of developing demyelination. By contrast, the risk of developing subsequent demyelination was only 8% when the barrier was intact. This strong association between barrier disruption and subsequent demyelination provides new insights into the role of blood brain barrier function in demyelinative disorders such as the osmotic demyelination syndrome and by extension to other demyelinative disorders such as multiple sclerosis. PMID- 10694843 TI - Polymorphisms of apolipoprotein E; outcome and susceptibility in multiple sclerosis. AB - Allelic variants of the apolipoprotein E (APOE) gene influence the course of several neurological diseases. In multiple sclerosis the concentration of APOE in cerebrospinal fluid and its intrathecal synthesis is reduced. Specific isoforms of APOE may also be important and it has been suggested that possession of the epsilon4 allele may be associated with a more aggressive disease process. These data prompted us to re-examine, in a large group of patients with multiple sclerosis, the proposal that allelism in the apolipoprotein gene influences disease course. Genotypes were determined in a well-defined group of 370 unrelated Caucasians with clinically definite multiple sclerosis and in 159 healthy controls. Age at onset, sex, disease duration, disease subtype were recorded. Disability was measured using the Kurtzke expanded disability status score in patients with a disease duration of 10 years or greater. There was no significant difference in APOE allele or genotype frequencies between patients and controls, between disease subtypes or between genders. APOE genotype did not significantly influence age of onset, and no significant relationship between genotype, allele frequency and disease severity was found. This study suggests that individual APOE alleles or genotypes do not determine disease susceptibility or the clinical course of multiple sclerosis. PMID- 10694844 TI - Macrophages: their myelinotrophic or neurotoxic actions depend upon tissue oxidative stress. AB - There are still questions regarding whether macrophages found in MS lesions are agents of recovery or of destruction. To address this, we examined in aggregate cultures prepared from dissociated embryonic spinal cord tissue, with or without addition of exogenous macrophages, the effect of menadione-induced oxidative stress. Similar to findings of other laboratories, we observed that in the absence of oxidative stress macrophage enrichment promoted myelinogenesis. In macrophage-poor cultures, menadione at 5 microM had very little effect upon the status of the aggregate cultures; however, increasing this to 10 and 20 microM did result in some damage to axons and myelin. By contrast, in macrophage enriched cultures, menadione at a concentration as little as 5 microM caused the complete destruction of the aggregates. We suggest that in neural tissues that have sufficiently high macrophage numbers, oxidative stress results in a positive inflammatory feedback loop that results in massive tissue destruction. We further suggest that what we see in macrophage-enriched aggregates subjected to oxidative stress may represent what happens in the Marburg-type of MS lesion. PMID- 10694845 TI - Contrast-enhanced magnetic resonance activity in relapsing remitting multiple sclerosis patients: a short term natural history study. AB - Magnetic resonance imaging (MRI) has been used to study the history of multiple sclerosis (MS). We analyze the relationship between MRI activity in the first scan compared to the subsequent five scans, and we evaluate whether a shorter observation period of 3 months may predict the subsequent 3 months. Monthly enhanced MRI was performed in 103 relapsing remitting (RR) MS patients for 6 months. Thirty-five per cent of patients had an inactive scan on the initial examination. More than 80% of them developed MRI activity during the following 5 months. Eighteen per cent of patients had three consecutive inactive scans; 65% of them had at least one active scan on the subsequent 3 monthly MRI's. The relationship between the first scan and all subsequent scans demonstrates a clear weakening over time. Eighty-two per cent of patients had at least one active scan during the initial 3 consecutive months, the chance of becoming inactive decreased from 23% to 0% over the subsequent 3 months, according with the mean number of enhancing lesions during the first 3 months. These results suggest that neither a single scan nor a short baseline of 3 months may adequately describe the natural history of disease in an individual RRMS patient. PMID- 10694846 TI - Cortical deficits in multiple sclerosis on the basis of subcortical lesions. AB - Patients suffering from multiple sclerosis have a high frequency of cognitive deficits usually attributable to demyelination and axonal loss in the subcortical white matter. Neurologic abnormalities referable to cortical function are uncommon but have been described. The present study describes three patients with clinically definite MS with deficits in cognitive function referable to cortical location. Two of the patients underwent positron emission tomography and showed profound cortical hypometabolism adjacent to subcortical white matter lesions seen on MRI. This paper points out that neurologic deficits referable to cortical sites may be caused by subcortical white matter lesions and that cognitive dysfunction in patients with MS may progress rapidly in the absence of motoria deficits or other evidence of clinical deterioration. Multiple Sclerosis (2000) 6 50 - 55 PMID- 10694847 TI - Drop in relapse rate of MS by combination therapy of three different phosphodiesterase inhibitors. AB - Phosphodiesterase inhibitors (PDEIs), when used in combination, synergistically suppress TNFalpha production by various cells and also suppress experimental demyelination at very low concentrations. We conducted a pilot study to determine whether the combination of three PDEIs suppresses the relapse of MS at the usual therapeutic doses. Of the 12 relapsing remitting MS, the mean relapse rate/year dropped remarkably (from 3.08+/-3.32 to 0.92+/-1.86) after PDEI treatment. Seven out of 12 (58.3%) were relapse-free in the follow up period (499+/-142 days). The combination of three PDEIs can be safe and useful strategy for the future treatment of MS. - 58 PMID- 10694874 TI - Clusterin is a secreted mammalian chaperone. PMID- 10694875 TI - Mapping chromosomal proteins in vivo by formaldehyde-crosslinked-chromatin immunoprecipitation. AB - Gene regulation is a complex process. Numerous factors appear to be required for the accurate temporal and spatial regulation of each gene. Often these factors are assembled into multiprotein complexes, contributing to specific gene regulation events. Understanding how all these factors are organized in the chromosome and how their function is regulated in vivo is a challenging task. One of the most useful techniques for studying this level of gene regulation is the in vivo fixation by formaldehyde crosslinking of proteins to proteins and proteins to DNA, followed by immunoprecipitation of the fixed material. PMID- 10694876 TI - Dennis chapman (1927-1999) PMID- 10694877 TI - Exonic splicing enhancers: mechanism of action, diversity and role in human genetic diseases. AB - Exonic splicing enhancers (ESEs) are discrete sequences within exons that promote both constitutive and regulated splicing. The precise mechanism by which ESEs facilitate the assembly of splicing complexes has been controversial. However, recent studies have provided insights into this question and have led to a new model for ESE function. Other recent work has suggested that ESEs are comprised of diverse sequences and occur frequently within exons. Ominously, these latter studies predict that many human genetic diseases linked to mutations within exons might be caused by the inactivation of ESEs. PMID- 10694878 TI - A superfamily of membrane-bound O-acyltransferases with implications for wnt signaling. PMID- 10694879 TI - SAP - a putative DNA-binding motif involved in chromosomal organization. PMID- 10694880 TI - Enzymes on the Web PMID- 10694881 TI - The dynamin family of mechanoenzymes: pinching in new places. AB - The large GTPase dynamin is a mechanoenzyme that mediates the liberation of nascent clathrin-coated pits from the plasma membrane during endocytosis. Recently, this enzyme has been demonstrated to comprise an extensive family of related proteins that have been implicated in a large variety of vesicle trafficking events during endocytosis, secretion and even maintenance of mitochondrial form. The potential contributions by the dynamin family to these diverse but related functions are discussed. PMID- 10694882 TI - Histone deacetylases: silencers for hire. AB - Over the past few years, the long-standing idea that covalent modification of chromatin can play a role in determining states of gene activity has been confirmed. Eukaryotic genes can be silenced by deacetylation of acetyl-lysine moieties in the N-terminal tails of histones. Recent work links histone deacetylases with an increasing number of repressors, suggesting that deacetylation might be a rather pervasive feature of transcriptional repression systems. PMID- 10694883 TI - Flavoenzymes: diverse catalysts with recurrent features. AB - Many biochemical processes exploit the extraordinary versatility of flavoenzymes and their flavin cofactors. Flavoproteins are now known to have a variety of folding topologies but a careful examination of their structures suggests that there are recurrent features in their catalytic apparatus. The flavoenzymes that catalyse dehydrogenation reactions share a few invariant features in the hydrogen bond interactions between their protein and flavin constituents. Similarly, the positioning of the reactive part of the substrate with respect to the cofactor is generally conserved. Modulation of substrate and cofactor reactivity and exact positioning of the substrate are key elements in the mode of action of these enzymes. PMID- 10694884 TI - Function of the ubiquitin-proteasome pathway in auxin response. AB - The plant hormone auxin regulates many aspects of growth and development. Despite the importance of this hormone, the molecular basis for auxin action has remained elusive. Recent advances using molecular genetics in Arabidopsis have begun to elucidate the mechanisms involved in auxin signaling. These results suggest that protein degradation by the ubiquitin pathway has a central role in auxin response. PMID- 10694885 TI - A novel FeS cluster in Fe-only hydrogenases. AB - Many microorganisms can use molecular hydrogen as a source of electrons or generate it by reducing protons. These reactions are catalysed by metalloenzymes of two types: NiFe and Fe-only hydrogenases. Here, we review recent structural results concerning the latter, putting special emphasis on the characteristics of the active site. PMID- 10694887 TI - NPS@: network protein sequence analysis. PMID- 10694886 TI - Eukaryotic DNA polymerases, a growing family. AB - In eukaryotic cells, DNA polymerases are required to maintain the integrity of the genome during processes, such as DNA replication, various DNA repair events, translesion DNA synthesis, DNA recombination, and also in regulatory events, such as cell cycle control and DNA damage checkpoint function. In the last two years, the number of known DNA polymerases has increased to at least nine (called alpha, beta, gamma, delta, epsilon, zeta, eta, t and iota), and yeast Saccharomyces cerevisiae contains REV1 deoxycytidyl transferase. PMID- 10694888 TI - Macromolecular interactions: tracing the roots. PMID- 10694889 TI - A Pseudomonas aeruginosa strain isolated from a contact lens-induced acute red eye (CLARE) is protease-deficient. AB - PURPOSE: Pseudomonas aeruginosa proteases are thought to be important virulence factors in the pathogenesis of corneal disease. This study examined protease production from two strains of P. aeruginosa responsible for two very distinct clinical diseases: strain Paer1, isolated from a Contact Lens-induced Acute Red Eye (CLARE), and strain KEI 1025, isolated from a corneal ulcer. Strains were compared to a laboratory strain (ATCC 19660) known to produce severe keratitis in experimentally infected mice for protease production and for ocular virulence. METHODS: Protease production was examined with colorimetric assays, gelatin zymography and western blots. Elastase A activity was quantitated with a staphylolytic assay. Ocular virulence was examined using a mouse scratch model of keratitis. RESULTS: In contrast to strains KEI 1025 or ATCC 19660, Paer1 was unable to produce enzymatically active elastase A, elastase, and protease IV. All three strains produced active alkaline protease. Strains KEI 1025 and ATCC 19660 produced a fulminant keratitis in mice whereas Paer1 produced a mild transient infection. Restoration of elastase activity in Paer1 via genetic complementation did not result in a virulent phenotype. Co-infection of mouse eyes with strains Paer1 and ATCC 19660 resulted in the eventual loss of Paer1 from corneal tissue. CONCLUSIONS: These studies suggest that P. aeruginosa elastase A and/or protease IV, but not alkaline protease or elastase, contribute to the ocular virulence of this organism. PMID- 10694890 TI - Growth regulation of bovine retinal pericytes by transforming growth factor-beta2 and plasmin. AB - PURPOSE: Transforming growth factor -beta2 (TGF-beta2) is a predominant isoform of TGF-betas in the eye and plasmin is a peptidase with many functions. To better understand the pathogenesis of retinal microcirculation disorders, the effects of TGF-beta2 and plasmin on cultured bovine retinal pericytes were investigated. METHODS: Exogenous TGF-beta2 or plasmin was added to some cultures, DNA synthesis during cell cycle progression was investigated using [(3)H]thymidine incorporation. Anti-TGF-beta2 antibody was added to neutralize the effects of TGF beta2. TGF-beta2 in the culture medium was measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Exogenous TGF-beta2 (10 pg to 100 ng/mL) suppressed DNA synthesis. Pericytes produced TGF-beta2. Anti-TGF-beta2 antibody neutralized TGF-beta2 and accelerated DNA synthesis, which shows that pericytes regulate their own cell cycle by action of the autocrine and/or paracrine system of TGF-beta2. Plasmin (0.2 to 0.5 U/mL) accelerated DNA synthesis in a dose dependent manner, while addition of aprotinin, a protease inhibitor, counteracted this effect of plasmin. The concentration of TGF-beta2 in the culture medium decreased with the addition of plasmin. Simultaneous addition of both plasmin and anti-TGF-beta2 antibody accelerated DNA synthesis. High and low glucose concentrations of the culture medium did not affect DNA synthesis. CONCLUSIONS: Our results suggest that TGF-beta2 and plasmin respectively decrease and increase DNA synthesis. In a retinal microcirculation disorder, they may play competitive roles in the cell cycle of pericytes. PMID- 10694891 TI - Growth factor mRNA and protein in preserved human amniotic membrane. AB - PURPOSE: To investigate the expression of growth factor mRNA and the level of growth factor protein in preserved human amniotic membrane (AM). METHODS: RT-PCR was used to examine the expression of mRNA for eight growth factors (EGF, TGF alpha, KGF, HGF, bFGF, TGF-beta1, -beta2, -beta3) and two growth factor receptors (KGFR and HGFR) in human AM preserved at -80 degrees C for one month. In addition, ELISAs were used to measure the protein concentrations of seven growth factors (EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2) in preserved human corneas and in AM both with and without amniotic epithelium. RESULTS: RT-PCR revealed that human AM expresses mRNA for EGF, TGF-alpha, KGF, HGF, bFGF, TGF beta1, -beta2, -beta3, KGFR and HGFR, while ELISAs showed that it contains EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2. AM without amniotic epithelium also contains all seven growth factors examined, however, in this tissue the protein levels of EGF, KGF, HGF and bFGF were found to be significantly lower than in native AM. CONCLUSIONS: Preserved human AM expresses mRNAs for a number of growth factors and contains several growth factor proteins that might benefit epithelialization after AM transplantation. High levels of EGF, KGF, HGF and bFGF in AM with amniotic epithelium as compared to AM without amniotic epithelium suggest an epithelial origin for these growth factors. We feel that EGF, KGF and HGF in particular might play important roles in ocular surface wound healing after AM transplantation. PMID- 10694892 TI - Lacrimal fluid peroxidase activity during the menstrual cycle. AB - PURPOSE: The aim of this work was to investigate peroxidase activity in human tears during the various phases of the menstrual cycle. For comparative purposes saliva was also examined. METHODS: Tear fluids and saliva from 10 healthy volunteers 23-41 years of age (mean: 28.2 years), with regular menstrual cycles were sampled for the duration of at least two complete cycles. Menstrual cycles and ovulation periods were evaluated by measuring morning body temperature and hormone levels in plasma and urine. Unstimulated tears and unstimulated saliva were collected in the morning every two days. Peroxidase activity was monitored according to the 5,5'-dithiobis, 2-nitrobenzoic acid thiocyanate (Nbs-SCN) method and the protein content was determined by the Bradford method. RESULTS: Peroxidase activity in tears, expressed as U/mL, was significantly (p <.05) higher during the preovulatory and luteal phases with respect to the menses, whilst total protein content remained almost unchanged throughout all phases. A positive correlation was found between lacrimal fluid peroxidase activity and 17beta-estradiol plasma levels (p <.001). Salivary peroxidase activity did not show such estrogen-related changes. CONCLUSIONS: Our findings report cyclic variations in peroxidase activity in human tears during the menstrual cycle. Such cycling seems to reflect variations of 17 beta-estradiol plasma levels. These results suggest that a regulation of lacrimal fluid peroxidase by 17 beta estradiol could be one possible cause for the female gender predilection in some ocular diseases, such as keratoconjunctivitis sicca. PMID- 10694893 TI - Lp85 calpain is an enzymatically active rodent-specific isozyme of lens Lp82. AB - PURPOSES: To clone and sequence the cDNA for Lp85 calpain from young rat lens, and to test for Lp85 protein expression and proteolytic activity. METHODS: RT-PCR and molecular cloning were performed on total RNA from 12 day-old rats. Lp85 protein expression was visualized by immunoblotting using a specific antibody developed to the unique peptide sequence in Lp85. Proteolytic activity was assessed by casein zymography. Transient expression of Lp85 and previously characterized lens-specific calpain Lp82 were separately performed in mammalian COS-7 cells. RESULTS: The 2410-bp cDNA for rat lens Lp85 encoded a protein of 737 amino acid residues with a calculated molecular weight of 85.0 kDa and a predicted pI of 5.67. The amino acid sequence of Lp85 was identical to Lp82 except for an insert region of 28 amino acids in domain IV of the calcium-binding region. mRNA and protein for Lp85 were present only in rat and mouse lenses and not in other tissues or species. Lp85 protein concentrations were highest in the nuclear region, most concentrated in the insoluble fraction, disappeared with lens maturation, and Lp85 exhibited migration similar to Lp82 on native PAGE gels. Lp85 was enzymatically active when expressed in COS-7 cells. CONCLUSIONS: Lp85 is a newly classified, lens- and rodent-specific, enzymatically active, member of the AX1 (alternative exon 1) subclass of calpains. In conjunction with Lp82 and m-calpain in lens, Lp85 may be responsible for proteolysis during normal lens development and maturation or during cataract formation in young rodents. PMID- 10694894 TI - Open-loop accommodation in emmetropia and myopia. AB - PURPOSE: To investigate the influence of method of measurement and refractive error on the open-loop accommodation response. METHODS: Open-loop accommodation was measured in darkness (dark accommodation, DA) and using a pinhole pupil (pinhole accommodation, PA) in emmetropic subjects (EMMs, n = 63), subjects with late-onset myopia (LOMs, n = 50) and subjects with early onset myopia (EOMs, n = 51). Further a control experiment examined the differences between DA and bright field accommodation (BA) conditions in a subset of subjects. All measurements of open-loop accommodation were carried out monocularly using a Canon R1 infra-red optometer in static recording mode. All myopic subjects were fully corrected using soft contact lenses. RESULTS: A significant variation (p < 0.001) in open loop accommodation was found between DA and PA, but no variation in open-loop level was observed between the three refractive groups. There was no interaction between these two factors. No significant difference was found between the BA level and DA level in any of the refractive groups. CONCLUSIONS: Open-loop accommodation response positions vary according to the experimental conditions employed during measurement. No refractive group differences in the open-loop response were apparent. PMID- 10694896 TI - Human retinal hemodynamics following administration of 5-isosorbide mononitrate. AB - PURPOSE: The purpose of this randomized double-masked cross-over study was to determine the effects of 20 mg of 5-isosorbide mononitrate (ISMO) on the retinal hemodynamics of young healthy subjects. METHODS: Monochromatic fundus photography and bidirectional laser Doppler velocimetry (BLDV) were used to determine vascular diameters (D), and blood velocity (V(max)) and flow (Q) in retinal veins, respectively. The diameter of the vein [D((vein))] at the same location where BLDV measurements were made, and the diameter of a neighboring artery [D((art) )], were determined from the fundus photographs. Measurements were carried out one and three hours after ISMO dosing, on twelve and six subjects, respectively. Mean blood pressure (BP(m)) and intraocular pressure (IOP) were also monitored, and ocular perfusion pressure (PP) was calculated. Results are expressed in percentage changes (+/- the standard error of the mean). RESULTS: On average, we observed a moderate increase of Q one hour after ISMO dosing (+8.2 +/ 5.4%), but not after placebo (+2.7 +/- 1.6%). This effect of ISMO, which displayed remarkable interindividual variability (95% confidence interval: -3.9%, +20.4%), did not attain statistical significance. D((vein)) and D((art)) were not appreciably affected. No effect was observed three hours after either ISMO or placebo dosing. PP was reduced one hour following ISMO administration, mainly as a function of reduced BP(m), although this variation was not statistically significant. IOP did not change appreciably throughout the duration of the study. CONCLUSIONS: Our findings suggest that, in contrast to the optic nerve head, in which we previously documented consistent and significant increases in blood flow following ISMO administration at both one and three hours, retinal hemodynamics are not equally responsive to a single dose of ISMO at these time points. Marked interindividual variability to the effects of this long-acting nitric oxide donor was documented one hour after administration, but not at three hours. This study further suggests that distinct vascular tissues of the ocular microcirculation respond differently to identical pharmacological challenges. PMID- 10694897 TI - Correlation of metalloproteinase-2 and -9 with proinflammatory cytokines interleukin-1b, interleukin-12 and the interleukin-1 receptor antagonist in patients with chronic uveitis. AB - PURPOSE: Matrix metalloproteinases have been shown to play a role in active uveitis. Transcription of MMPs is induced by a number of growth factors and cytokines. This study investigates the role of MMPs in chronic uveitis and correlates the amounts of MMP-2 and -9 in intraocular samples to the intraocular levels of proinflammatory cytokines (Interleukin-1 [IL-1], Interleukin-12 [IL-12] and Interleukin-1 receptor antagonist [IL-1ra]). METHODS: Aqueous humor of 16 patients was collected during surgical or diagnostic procedures (control group: cataract patients). MMP-2 and -9 were measured using zymography. IL-1beta, IL-12 and IL-1ra were evaluated by ELISA. RESULTS: We found MMP-2 and -9 in all of our uveitis patients. In the control group only MMP-2 was seen. Higher levels of MMP 2 and -9 were found in patients with higher activity of uveitis (p < 0.014 for MMP-2, p < 0.0054 for MMP-9). The amounts of IL-1beta, IL-12 and IL-1ra detected in our uveitis patients correlated with levels of MMP-2 (p < 0.07, p < 0.0004, p < 0.03) and MMP-9 (p < 0. 003, p < 0.0001, p < 0.002), and IL-12 (p < 0.004, p < 0.0001). Patients with moderately active uveitis presented with twice the level of MMP-2 as the control group; MMP-9 levels reached up to 92% of the amounts found in patients with active uveitis. Two patients in remission for almost one year still had detectable intraocular MMP-9 levels. CONCLUSION: Our data suggests that the high levels of MMPs found in patients with chronic uveitis might contribute to the damage often seen in these eyes. Since MMPs are capable of releasing proinflammatory cytokines bound to components of the extracellular matrix, and facilitate the secretion of active TNF-alpha by cleavage of the membrane bound form, it is conceivable that MMPs contribute to the chronicity of some uveitis cases. PMID- 10694895 TI - Expression of the extraneuronal monoamine transporter in RPE and neural retina. AB - PURPOSE: Dopamine has several important functions in the retina including a possible role in controlling photoreceptor disk shedding to the RPE. While some cells express a transporter for dopamine, the RPE cell does not, leading us to ask whether the newly described catecholamine transport system, the extraneuronal monoamine transporter (uptake(2)) (also known as organic cation transporter 3 (OCT3), is present in RPE and might function as a transporter for dopamine. OCT3 also accepts histamine as a transportable substrate and so we investigated the interaction of this retinal neurotransmitter with OCT3. METHODS: OCT3 expression in the mouse eye was analyzed by in situ hybridization, Northern blot analysis and RT-PCR. OCT3 function was analyzed in cultured human ARPE-19 cells by monitoring the uptake of 1-methyl-4-phenyl pyridinium (MPP(+)), a neurotoxin, which is a known substrate for OCT3. RESULTS: In situ hybridization analysis showed that OCT3 is expressed in mouse RPE and in several cell types of the neural retina, including photoreceptor, ganglion, amacrine, and horizontal cells. The expression of OCT3 in RPE was confirmed by Northern blot analysis and RT-PCR. The characteristics of MPP( +) uptake in cultured ARPE-19 cells included the stimulation of transport by alkaline pH, high affinity (K(t) = 28 +/- 4 microM), competition with several cationic drugs and monoamine neurotransmitters and sensitivity to steroids. In addition, the uptake of MPP(+) in RPE cells was inhibited by dopamine and histamine with IC(50) values (concentration needed for 50% inhibition) of 637 +/- 84 microM and 150 +/- 20 microM, respectively. CONCLUSIONS. This study provides the first report on the expression and function of an organic cation transporter, OCT3, in the eye and in particular the RPE. The data have physiological and pharmacological relevance as it is likely that OCT3 participates in the clearance of dopamine and histamine from the subretinal space and may also play a key role in the disposition of the retinal neurotoxin MPP(+). PMID- 10694898 TI - Genistein produces reduction in growth and induces apoptosis of rat RPE-J cells. AB - PURPOSE: To investigate the effect of the tyrosine kinase inhibitor, genistein, on the growth of the retinal pigment epithelial (RPE) cell. METHODS: The tyrosine kinase inhibitor, genistein, was administered in culture to the rat retinal pigment epithelial cell line, RPE-J. The effect on cell viability and growth was assessed by trypan blue dye exclusion. Induction of apoptosis was confirmed morphologically by light and electron microscopy and oligonucleosomal fragmentation was assessed by TUNEL and DNA ladder. Quantitation was undertaken by propidium iodide staining and photometric enzyme immunoassay. Western blot was performed to study poly-(ADP-ribose)-polymerase cleavage (PARP). To confirm the involvement of caspase, the caspase inhibitor z-VAD-fmk was employed. In addition, cell cycle phase was determined by flow cytometry. RESULTS: We here demonstrate that genistein treatment of RPE-J cells produces a dose- and time dependent growth inhibition. Genistein in higher concentration induces apoptosis of rat RPE-J cell. z-VAD-fmk inhibited this type of apoptosis and cleavage of PARP enzyme was demonstrated. Ten micromolar genistein inhibited cell proliferation by G(0)/G(1) arrest without inducing apoptosis of the major population. Whereas 50 microM genistein caused growth inhibition of RPE-J cells by G(2)/M arrest and subsequent apoptotic death. CONCLUSIONS: Genistein inhibits RPE cell growth and induces apoptosis. The ability of genistein to inhibit the proliferation and to induce apoptosis of RPE cells could be potentially therapeutic for proliferative vitreoretinopathy. PMID- 10694899 TI - Effect of melatonin against oxidative stress in ultraviolet-B exposed rat lens. AB - PURPOSE: To investigate the defensive effect of melatonin against oxidative stresses in ultraviolet-B (UVB) radiation induced cataract development. METHODS: Young rats received 8 kJ/m(2) UVB for 15 min. For the intervention of cataract development intraperitoneal injection of melatonin (4 mg/kg daily for 1 week) following UVB exposure was performed. Lenticular glutathione peroxidase (GSHPx), catalase (CAT), superoxide dismutase (SOD) activities and glutathione (GSH), malondialdehyde (MDA) levels were determined in UVB-melatonin, UVB, and control groups. RESULTS: One week after exposure in the UVB group, lens opacities were observed and CAT, SOD, and GSHPx activities, and GSH level were lower than control and MDA level was higher than control (p < 0.05). In the UVB-melatonin group CAT and SOD activities were lower than control (p < 0.05), and the MDA level was lower than the UVB group (p < 0.05). CONCLUSIONS: These results suggest that melatonin may protect against the UVB-induced cataract development by directly quenching lipid peroxides and indirectly by enhancing the production of the endogenous antioxidant GSH. PMID- 10694900 TI - Morphologic preservation and variability of human donor retina. AB - PURPOSE: To facilitate studies of human retina and utilization of human retinal tissue for treatment of retinal diseases, we studied morphologic preservation in postmortem human retina. METHODS: Morphology of retinas from thirty-one human eyes was examined using light and electron microscopy. The inner and outer retina, rod and cone photoreceptor cells, and central and peripheral retina were compared with regard to morphologic preservation. Possible factors affecting survival were analyzed. RESULTS: The earliest postmortem change was vacuolation of the nerve fiber layer within a few hours postmortem, followed by vacuolation and cytoplasmic swelling of the inner retina. As compared with the inner retina, outer retinal structure was better preserved, i.e., the photoreceptor cells maintained better morphology. Rod cell morphology was better preserved than cone cell morphology, with good preservation of the rod outer segment disc membranes and the inner segment mitochondrial membranes. Thus, well-preserved rod photoreceptor cells were evident in specimens at least 48-hours postmortem. Peripheral retina was better preserved than the central retina including the fovea and perifovea. Factors affecting anatomical integrity included the total time postmortem and, more importantly, the time between death and enucleation. Other factors, including age and sex, did not appear to affect morphological preservation in the present study. CONCLUSIONS: Human retina postmortem remained morphologically intact for a relatively long period of time, with differential preservation among different geographic areas and cell types. This morphologic evidence is consistent with previous findings of functional preservation (e.g. , photoresponses) in such tissue. This study may shed some light on understanding of human retina and its utilization for retinal transplantation. PMID- 10694901 TI - Molecular identification of functional water channel protein in cultured human nonpigmented ciliary epithelial cells. AB - PURPOSE: Water channel proteins are important pathways for water movements across cell membranes, including those in the ciliary epithelium, which is the major site of aqueous humor secretion. In this study, we aimed to demonstrate the expression of functionally active aquaporin-1 (AQP1) water channels in cultured human ciliary epithelial cells. METHODS: Poly A(+) RNA was isolated from cell cultures of Simian Virus 40 (SV-40) transformed human nonpigmented ciliary epithelium (NPE) subjected to RT-PCR reaction using primers specific to AQP1. Northern analysis was used to define the expression of AQP1 in NPE cells. Western immunoblotting with polyclonal antibody raised against AQP1 was used to evaluate the AQP1 protein expression in the plasma membranes of human NPE cells. Light scattering method was used to determine the osmotic water permeability in the suspension of NPE cells. RESULTS: RT-PCR using specific primers for AQP1, Northern analysis and Western immunoblot using AQP1 specific antibody demonstrated the expression of AQP1 in the plasma membranes of NPE cells. Osmotic water permeability (P( f)) measurements confirmed that functional AQP1 water channels are expressed in human NPE cells and the P(f) for these cells was 9.8 x 10( -3) cm/s at 10 degrees C. CONCLUSIONS: The presence of AQP1 in human NPE cells suggests that it may have a role in the fluid flow across epithelial membranes. In addition, the existence of AQP1 in the human NPE cells provide an excellent in vivo model to study the regulation of aquaporins and their possible role in the aqueous humor secretion. PMID- 10694902 TI - Inter-ocular characteristics of the pre-contact lens tear film. AB - PURPOSE: To establish whether the status of the pre-contact lens tear film as indicated by standard, clinical observational techniques is affected by moderate stimulation of the contralateral eye. METHODS: Four indicators of tear film behavior, lipid layer appearance, amount of debris, inferior meniscus height and non-invasive tear break up time (NIBUT) were monitored in ten subjects before and during 30 minutes of monocular pHEMA contact lens wear. Concurrently, the contralateral eye was either subjected to moderate irritation by means of a silicone elastomer contact lens, or remained unstimulated. Data were compared, between the stimulated and unstimulated states to identify evidence of contralateral treatment effects. RESULTS: After 30 minutes, maximum contralateral differences between the unstimulated and stimulated conditions were 1 grade for both lipid layer appearance and debris, 0.1 mm for meniscus height and 4 secs for NIBUT. CONCLUSIONS: The magnitudes of contralateral effects induced by moderate, monocular irritation were comparable with the within-subject variabilities associated with these indicators of tear film behavior. PMID- 10694903 TI - Ultrastructural histochemical studies of secretory processes in rat submandibular granular tubules during intermittent sympathetic nerve stimulation. AB - Secretory changes in the cells of granular tubules in rat submandibular glands have been studied sequentially during electrical stimulation of their sympathetic nerves. Results were assessed in a series of biopsied lobes from the same gland, taken at different times during the sympathetic stimulation. Changes were not synchronous between adjacent cells and it appeared that the time for the onset of secretory events differed between cells but, once set in action, a chain of similar events occurred. Nevertheless, some cells appeared to remain refractory throughout. Initially, some alignment of granules to the adjacent plasma membrane occurred and occasional evidence for classical exocytosis was seen. However, from early on microvesicles appeared in more luminally located granule membranes and were associated with granule fusions, that became common and led to the formation of large irregular aggregates. Most of the secretion of granule contents appeared to be through openings of aggregates into lumina. With granule fusions the intra membrane microvesicles became internalised and tended to increase in size with time; they were commonly expelled with the contents of the aggregates. Fragments of cytoplasm also became incorporated in aggregate formation. Cytoplasm, often containing glycogen, also formed luminal blebs over some granular tubule cells and appeared to pass into the secretion by an apocrine process. At the end of stimulation multivesicular bodies were seen in association with redundant aggregates. PMID- 10694904 TI - Changes in numbers and dimensions of glomeruli during metamorphosis of Pelobates syriacus (Anura; Pelobatidae). AB - Changes in glomerular numbers and volume were followed throughout five ontogenetic stages of Pelobates syriacus (Anura, Pelobatidae). The number of glomeruli increased markedly between the 2-legged and the 4-legged tadpole stages. In the post-metamorphic stages the number of glomeruli was about twice their number during the tadpole stages. Glomerular volume peaked during the 2 legged tadpole stage, when the tadpole reached its maximal size, and dropped thereafter. There is some evidence to indicate that the number of glomeruli in post-metamorphic P. syriacus increased during growth thus indicating perhaps an increase in their functional importance. The number was still lower than that reported for some other anurans, more similar to the numbers given for representative urodeles, however, the latter have considerably larger glomeruli. Two main factors appear to be involved in regulating numbers and volume of glomeruli in frogs. The ontogenetic changes during metamorphosis affect both glomerular numbers and volume, whereas post-metamorphic growth affected only the glomerular numbers. PMID- 10694905 TI - Effect of genistein on the temporal coordination of cleavage and compaction in mouse preimplantation embryos. AB - Initially, we investigated the effect of genistein, an inhibitor of protein tyrosine kinases, on compaction of the mouse embryo since tyrosine phosphorylation of the cadherin-catenins complex was suggested to down-regulate its adhesive function. Genistein prevented cleavage from the 2- to the 4-cell stage in a concentration-dependent manner. The next cleavage is inhibited at all concentrations used. Time course of intercellular flattening is however identical for both control 8-cell embryos and 4-cell arrested embryos. This confirms that compaction takes place according to a biological clock that does not depend on completion of the third cell cycle. Our results also suggest that, since, in contrast to genistein, protein kinases C modulators are known to cause a premature compaction, diacylglycerol-dependent kinases but not protein tyrosine kinases might be upregulators of compaction. PMID- 10694906 TI - The Harderian gland of the Cheesman's gerbil (Gerbillus cheesmani ) of the Kuwaiti desert. AB - The Harderian gland is a large orbital structure. Several functions have been ascribed to the gland such as lubrication of the eye, a source of pheromones, thermoregulartory lipids and photoprotective secretions and a part of the retinal pineal axis. In the present study, the Harderian gland of the Cheesman's gerbil, Gerbillus cheesmani, is described for the first time. The gland is located around the posterior portion of the eyeball. The gland is compound tubular, surrounded by a thin connective tissue capsule. Only one secretory epithelial cell type was recognized, characterized by the presence of lipid vacuoles and cytoplasmic slashes in high numbers; the former being more concentrated towards the apical part while the latter being more concentrated towards the central and basal parts. Some of the cytoplasmic slashes contained electron dense filamentous structures. Similar structures were observed in the lipid vacuoles. Thus, a functional relationship between the cytoplasmic slashes and the lipid vacuoles is suggested. A unique structure was observed, termed dome-like cells, located between the epithelial cells and the basement membrane. These cells were characterized by the extensive presence of pleomorphic mitochondria and compact lamellae of granular endoplasmic reticulum (GER) in the form of finger prints. The gland was found to be actively secreting porphyrins as well as lipids. Cellular debris was also seen in the tubular lumina. Myoepithelial cells with their spindle shape and elongated nuclei were evident between the basement membrane and the secretory epithelium. Sparse interstitial tissue was observed in between the gland tubules of both male and female gerbils. Macrophages, dendritic melanocytes and lymphocytes are the most represented cellular components of the interstitium. Further studies are required to investigate the function of the dome-like cells as well as the role of lymphocytes in the rodents Harderian gland. PMID- 10694907 TI - Microwave processing for scanning electron microscopy. AB - The normal processing of biological samples for Scanning Electron Microscopy, includes treatment with aldehyde (1 to 2 hours), postfixation with Osmium (1 hour), followed by dehydration in a ascending grade of ethanol (30 a 100%), 10 to 15 minutes in each step, and finally drying. This procedure takes at least 8 hours. In this work, samples of mosquitoes (Aedes), protozoa (Tritrichomonas muris), bacteria (Clostridium oceanicum), murine liver, and small intestine were processed in the same manner in a domestic microwave oven for two minutes at 20% of its maximum power. The complete procedure from the initial fixation to dehydration in 100% ethanol was reduced to one hour with good preservation of the ultrastructural details of the specimens. PMID- 10694908 TI - Mechanical properties and chemical composition of avian long bones. AB - We have studied the mechanical behaviour of avian long bones as whole structures, by calculating mechanical parameters such as maximum load, stiffness, bending strength and flexural Young's modulus; bones were always tested in three-point bending. Furthermore composition in several chemical elements and amino acids related to collagen content was also analysed. Correlations were established between body mass, mechanical parameters and chemical contents. Both bending strength and Young's modulus were negatively correlated to body mass. Significant correlations were found between nitrogen content and both strength and Young's modulus, with negative slopes in both cases. Magnesium and phosphorus appear to be the most important inorganic elements to the understanding of the mechanical behaviour of avian long bones. PMID- 10694909 TI - Closure of the uterine lumen and the plasma membrane transformation do not require blastocyst implantation. AB - Ultrastructural changes in the plasma membrane of uterine epithelial cells in the pseudopregnant rat were examined to determine if these changes resemble those found during normal pregnancy and also to examine if the well-known membrane alterations of early pregnancy are intrinsic to uterine epithelial cells. Changes in the surface contours of uterine epithelial cells from the afternoon of day 6 to the morning of day 9 of pseudopregnancy were similar to those present after attachment in normal pregnancy although somewhat delayed. The presence of short, irregular microvilli was seen from as early as day 7 of pseudopregnancy, with regular microvilli returning to the epithelial surface by days 8-9 of pseudopregnancy but to a slightly lesser extent as compared to normal pregnancy. Furthermore, observations made on the afternoon of day 6 to the morning of day 7 of pseudopregnancy showed that the uterine lumen was closed down and that complete membrane flattening between opposing uterine epithelial cells was seen all along the uterus in the absence of a blastocyst. These observations establish that the "plasma membrane transformation" does not depend on blastocyst implantation. PMID- 10694910 TI - The relationship of callosal anatomy to paw preference in dogs. AB - Previous studies have described the paw preference and asymmetry in dog brains, based on experimental studies. The purpose of the present study is to investigate a possible association between callosal anatomy and paw preference in dogs. The midsagittal area of the dog corpus callosum was measured in its entirety and in six subdivisions in a sample of 21 brains obtained from 9 male and 12 female mongrel dogs which had paw preference testing. The present study showed significant paw differences in dog corpus callosum. A posterior segment of the callosum, the isthmus, was significantly larger in the right pawedness than the left. PMID- 10694911 TI - Electron microscopic contrast of the cytoskeleton and junctional complexes of intestinal epithelial cells by ethanolic phosphotungstic acid. AB - After glutaraldehyde fixation and treatment with ethanolic phosphotungstic acid (E-PTA) before plastic embedding, sections of rat large intestine showed a characteristic electron contrasting pattern in epithelial cells. The axis of microvilli, terminal web, a thin band below the luminal plasma membrane, centrioles and junctional complexes (tight junctions, adherens junctions, and desmosomes) appeared highly contrasted. In addition to protein components of microfilaments and intermediate filaments, proteins from the junctional complexes could also be implicated in the contrasting reaction with E-PTA. Mitochondrial membranes, chromatin masses, and nucleoli of enterocytes showed considerable electron density, whereas no reaction was found in the glycocalyx and mucin content of goblet cells. The clear visualization of cytoskeleton elements and junctional complexes by E-PTA contrasting represents a simple and valuable method for studies on the normal and pathological organization of these structures in epithelial cells. PMID- 10694912 TI - Effect of infantile strabismus on visuomotor development in the squirrel monkey (Saimiri sciureus): optokinetic nystagmus, motion VEP and spatial sweep VEP. AB - PURPOSE: To determine whether squirrel monkeys made artificially strabismic in infancy had ocular fixation abnormalities, directional asymmetries of horizontal optokinetic nystagmus (OKN) and asymmetries of motion visually evoked potentials (MVEPs) similar to those of humans with infantile strabismus. METHODS: Esotropia was produced in a newborn squirrel monkey by surgical tenotomy of both lateral rectus muscles. The alignment and eye rotations of the monkey were examined longitudinally and VEP testing was performed at the age of one year. Visual acuity was measured using spatial frequency sweep VEPs (SSVEP) in response to grating stimulation. OKN was tested under conditions of monocular viewing using full-visual-field, vertically oriented, moving stripes. MVEPs in response to horizontal motion were recorded with the animal sedated to reduce the possibility of eye movement artifact. RESULTS: The artificially strabismic squirrel monkey displayed a constant, committant esotropic strabismus accompanied by latent nystagmus. Monocular SSVEP acuity was subnormal in one eye, consistent with mild monocular strabismic amblyopia. The monkey demonstrated asymmetric OKN favoring nasally-directed stimulus motion when viewing with either eye. Monocular MVEPs were also characterized by a horizontal asymmetry with a directional bias inverted 180 degrees between the right and the left eyes. The eye movements and MVEP asymmetries were similar to those observed in strabismic macaque monkeys and humans with early-onset strabismus. Neither the OKN asymmetry nor the MVEP asymmetry was evident in a normal squirrel or normal macaque monkey. CONCLUSION: The artificially strabismic squirrel monkey is an appropriate eye movement and VEP model for the study of neural mechanisms in human infantile strabismus. PMID- 10694913 TI - Effect of botulinum toxin injections into rabbit eye. AB - The aim of this study was to investigate the adverse effects of the intraocular injection of botulinum toxin in rabbits. Intravitreal injections of botulinum toxin A in five doses, 1.25, 2.5, 5, 10, 25 units, were given into five rabbit eyes. The same volume of saline was injected into the second eye of the rabbit as a control. External examination, ophthalmoscopy, visual evoked potentials and electroretinography were done before injection and repeated at the first and second weeks after the injection. There were no significant differences in retinal function between toxin- and saline-injected eyes, neither ophthalmoscopically nor electrophysiologically. Ipsilateral mydriasis developed in the eyes injected with botulinum toxin. This study suggests that botulinum toxin has no harmful effect on retinal function. PMID- 10694914 TI - Results of classical and augmented bimedial rectus recession in infantile esotropia. AB - PURPOSE: To compare the success rates of augmented bimedial rectus recession and the standard recession. MATERIALS AND METHODS: Ninety patients were included in the study. The patients were evaluated in two groups according to the amount of recession. Group 1, the standard surgery group, received 5 mm or less of recession; Group 2, the augmented surgery group, received 6 mm of recession or more. The mean postoperative follow-up was 29 months (6-60 months) in Group 1, and 20 months (6-58 months) in Group 2. RESULTS: The mean age at the time of surgery was 4.61 years in Group 1 and 4.58 years in Group 2. The 56 patients in Group 1 underwent bilateral rectus recession varying from a minimum of 3 mm to a maximum of 5 mm; the 34 patients in Group 2 had recessions varying from a minimum of 6 mm to a maximum of 8 mm. The mean preoperative angle size was 39.64 +/- 8.93 SD (range 20-50 PD) in the standard surgery group, and 59.70 +/- 10. 04 SD (range 51-85 PD) in the augmented surgery group. The average postoperative deviation was 13.37 +/- 11.87 SD (range 0-45) in Group 1 and 9.02 +/- 10.02 (range 0-45) in Group 2. A good surgical result was achieved with one operation in 29 of 56 patients (51.8%) in Group 1 and 24 of 34 patients (70.58%) in Group 2. DISCUSSION: The optimal surgical technique for the correction of large-angle esotropia is still controversial; it appears that the augmented bilateral medial rectus recession is an effective and reasonable alternative to three- or four muscle procedures as the initial surgical treatment. PMID- 10694915 TI - The role of vergence adaptation in recovery of binocular single vision (BSV) following sensory strabismus. AB - Vergence adaptation is an important element of comfortable binocular single vision and probably contributes to the high incidence of orthophoria or small angles of heterophoria in the normal population. Where binocular single vision has been absent for a period of time, restoration of good visual acuity appears to enable the vergence adaptation mechanism to become active again. A case is presented in which a moderate to large angle of deviation rapidly 'disappeared' once good visual acuity was restored and the disparate images could be fused. Known factors concerning vergence adaptation are discussed in relation to such clinical cases. PMID- 10694916 TI - Novel compound heterozygous laminina2-chain gene (LAMA2) mutations in congenital muscular dystrophy. Mutations in brief no. 159. Online. AB - The laminina2-chain gene (LAMA2) encodes a basal lamina protein, laminina2, known to be deficient in one form of congenital muscular dystrophy (CMD). In a laminina2 deficient-CMD patient, we screened the entire LAMA2 cDNA (953bp) by reverse transcriptase polymerase chain reaction combined with single strand conformational polymorphism analysis. Direct sequencing of aberrant conformers in this patient revealed two loss-of-function mutations, consistent with autosomal recessive inheritance. The patient had two novel heterozygous mutations: 1) an exon 4 nonsense mutation caused by a G-->A substitution at cDNA position 547, changing the TGG codon for tryptophan into a TGA stop codon (W166X) in the N terminus domain VI;ii) an exon 54 frameshift mutation due to a deletion of nucleotide 'C' at cDNA position 7707 (S2553Y), resulting in a premature stop codon (V2587X) in exon 55 in the globular G domain of laminina2 at the C terminus. These mutations cause a disruption of the open reading frame of LAMA2. The absence of laminina2 observed in the patient's muscle biopsy could result from diminished levels of the LAMA2 transcript. Alternatively, the mutations might lead to translation of a truncated laminina2. By either mechanism the phenotype of congenital muscular dystrophy is believed to be the result of disruption of linkage between the extracellular matrix and the dystrophin glycoprotein complex. PMID- 10694917 TI - Transthyretin Ile73Val is associated with familial amyloidotic polyneuropathy in a Bangladeshi family. Mutations in brief no. 158. Online. AB - Amyloidosis is characterised by the extraceullular deposition of certain different proteins in a distinctively abnormal fibrillar conformation. All types of amyloid fibril share remarkably similar structural and biophysical properties despite substantial chemical heterogeneity among their respective precursor proteins. Hereditary amyloidosis associated with genetically determined protein variants is rare, but is extremely important as a model for studying the pathogenesis of amyloidosis generally. We report a novel mutation of the transthyretin (TTR) coding for TTR Ile73Val which is associated with familial amylodotic polyneuropathy (FAP) in a Bangladeshi family. The mutation was detected by direct sequencing of the PCR-amplified TTR exons. It creates an additional Accl restriction exzyme site in exon 3, allowing confirmation of its presence by RFLP. Amyloid detected in sural nerve and colonic biopsies was shown to be composed of TTR by immunohistochemistry. The predominant clinical features were progressive autonomic and sensori-motor peripheral neuropathy, beginning at age 50 years. The proband's father and two siblings had similar illnesses. These findings indicate Val73 is an amyloidogenic variant of TTR. PMID- 10694918 TI - A nonsense mutation (R242X) in the branched-chain alpha-keto acid dehydrogenase E1alpha subunit gene (BCKDHA) as a cause of maple syrup urine disease. Mutations in brief no. 160. Online. AB - Mutation analysis of DNA from cultured amniocytes with absent branched-chain alpha-ketoacid dehydrogenase activity revealed a C to T transition producing a nonsense mutation (R242X) in exon 7 of the gene encoding the E1a subunit of this multienzme complex (BCKDHA). This pregnancy occured in a large consanguinous pedigree with mutiple individuals with maple syrup urine disease (MSUD). PCR amplification of the region surrounding exon 7 allowed the identification of this mutation as well as two other previously identified mutations which cause MSUD. PMID- 10694919 TI - Niemann Pick Disease type A in Israeli Arabs: 677delT, a common novel single mutation. Mutations in brief no. 161. Online. AB - A novel single base pair deletion in the acid sphingomyelinase (ASM) gene (677delT in the cDNA) was identified in 12 Israeli Arab families with Niemann Pick disease (NPD) type A. This deletion creates a premature stop codon which explains the complete deficiency of ASM activity in these patients and the severe clinical manifestation. A single mutation in 12 families living in a relatively small geographical region suggests a founder effect and explains the high frequency of this disease in this population. This is in contrast to multiple mutations found in two other lysosomal storage disorders prevalent in this population, namely, Hurler disease (MPSI) and metachromatic leukodystrophy. Mutations analysis is therefore an important tool in characterizing the grounds for the high frequency of inherited diseases as well as a basis for prevention programs for prevalent diseases through carrier identification and the ascertainment of high risk families. PMID- 10694920 TI - Three novel missense mutations in the glucokinase gene (G80S; E221K; G227C) in Italian subjects with maturity-onset diabetes of the young (MODY). Mutations in brief no. 162. Online. AB - The maturity-onset diabetes of the young (MODY), an autosomal dominant form of non-insulin dependent diabetes mellitus (NIDDM), is caused by mutations in the glucokinase (GK, MODY 2) and in the hepatocyte nuclear factor 1a (MODY 3) and 4a (MODY 1) genes. We have screened the glucokinase gene by the polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) in fifteen subjects with clinical characteristics of MODY and one parent with NIDDM, impaired glucose tolerance or gestational diabetes. PCR products with abnormal mobility in DGGE were directly sequenced. We have identified four mutant alleles, three of them (G80S, E221K, G227C) are new missense mutations located in or near the region of the active site cleft of the enzyme. The mutations co-segregate with hyperglycemia in the families of the three probands, whose biochemical and clinical phenotype is similar to other individuals with MODY 2 mutations. PMID- 10694921 TI - A novel mutation in the neonatal region of the fibrillin (FBN)1 gene associated with a classical phenotype of Marfan syndrome (MfS). Mutations in brief no. 163. Online. AB - Marfan Syndrome (MfS) is an autosomal dominant inherited connective tissue disorder with variable phenotypic expression of cardiovascular, skeletal and ocular manifestations. Cardiovascular complications, such as aortic aneurysm and dissection drastically reduce life expectancy of individuals with MfS, whereas preventive surgery substantially improves the prognosis of these patients. A number of mutations in the fibrillin 1 (FBN1) gene associated with MfS have been identified to date, demonstrating considerable molecular heterogeneity. One region, however, located around exon 24, exhibits a striking clustering of mutations, which are associated with a severe, socalled neonatal form of MfS. Here we report the first mutation (G2950A) in exon 24 of the neonatal region of the FBN1 gene, associated with a classic MfS phenotype. The mutation leads to the subsitution of valin by isoleucin (V984I), both uncharged amino acids, which only differ in a single methyl group. This defect was identified in a proband with cardiovascular manifestations of MfS by SSCP analysis of PCR-amplified genomic DNA, direct PCR sequencing and RFLP analysis. The substitution was neither detected in the unaffected 4-year old daughter of the proband, nor in 3 of his healthy family members nor in 108 allels from control individuals, suggesting that this mutation is causative for MfS in the patient. Since no other family member of the proband is affected by MfS, the defect described is sporadic. In summary, we identified a novel defect in exon 24 of the neonatal region of the FBN1 gene in a patient with a classic phenotype of MfS, suggesting that conservative substitutions in this region may lead to a less severe phenotype of the disease. This finding further demonstrates the remarkable phenotypic heterogeneity associated with FBN1 mutations and stresses the significance of modifying genes and individual alterations in protein function for the pheontypic expression of the disease. PMID- 10694922 TI - Novel missense and frameshift mutations in the activin receptor-like kinase-1 gene in hereditary hemorrhagic telangiectasia. Mutations in brief no. 164. Online. AB - Hereditary hemmorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterized by multisystemic vascular dyplasia and recurrent hemorrhage. One of the causative genes is the activin receptor-like kinase-1 (ALK-1) gene located on chromosome 12q13. ALK-1 is an endothelial cell type I receptor for the TGF-beta superfamily of ligands. As a number of mutations have been identified in the kinase domain of ALK-1, we initiated a mutation analysis specifically targeting the first four coding exons of ALK-1 in order to determine if mutations in the extracellular and transmembrane domains are also present in HHT. Six new mutations have been identified. Three frameshift mutations were identified in exons encoding the extracellular and transmembrane domains. These mutations would grossly truncate the ALK-1 protein and are thus classic null alleles. Three new missense mutations within the exons encoding the extracellular domain, in addition to two previously described missense mutations, are located at or near highly conserved cysteines. These mutations may disrupt intra- or inter-molecular disulfide bridges required for ligand binding. The combined data suggest that both severe and subtle changes in the ALK-1 amino acid sequence can lead to receptor dysfunction and result in the HHT disease phenotype. PMID- 10694923 TI - C112R, W323S, N317K mutations in the vasopressin V2 receptor gene in patients with nephrogenic diabetes insipidus. Mutations in brief no. 165. Online. AB - Nephrogenic diabetes insipidus (NDI) is a rare, mostly X-linked recessive disorder characterized by renal tubular resistance to the antidiuretic effect of arginine vasopressin. The gene responsible for the X-linked NDI, the G-protein coupled vasopressin V2 receptor, has been localized on the Xq28 region. In this study we present three NDI families from Hungary with three different missense mutations in the vasopressin V2 receptor gene. After the mutations in the affected probands in each family had been characterized, other family members were screened by restriction enzyme analysis. The N317K and W323S mutations have not been detected previously. The C112R is an already known mutation. The N317K was a de novo mutation in the patient. The C112R and the W323S were found in the mothers of the patients as carriers and in all other patients, but not in the unaffected members of the families. Segregation of the mutations was consistent with the clinically observed symptoms as well as their severity. As conclusion, these findings further evidence that X-linked NDI results from defects in the V2 receptor gene. PMID- 10694924 TI - Identification of a new heterozygous point mutation in the COL1A2 gene leading to skipping of exon 9 in a patient with joint laxity, hyperextensibility of skin and blue sclerae. Mutations in brief no. 166. Online. AB - A heterozygous deletion of exon 9 in the COL1A2-mRNA of a patient with symptoms of both the Ehlers-Danlos-Syndrome and the Osteogensis Imperfecta is described. In the genomic DNA of the patient, exon 9 is homozygously present. We identified a novel heterozygous point mutation in the splice donor site of intron 9, leading to a G-->A substitution in position +5. This mutation leads to heterozygous skipping of exon 9 in the COL1A2-mRNA of this patient. The deletion results in a shortened (by 18 amino acids) but in frame 12(1) chain, which probably leads to the formation of abberantly processed triple helices. PMID- 10694925 TI - Identification of novel PAX6 mutations in two families with bilateral aniridia. Mutations in brief no. 167. Online. AB - We report two novel PAX6 mutations in aniridia patients of two Swiss pedigrees (We, Sc) which give rise to different phenotypes. An SSCP analysis of the PAX6 14 exons reveals electrophoretic mobility shifts exclusively in exons 5 and 12 of aniridia patients. As determined by bidirectional sequencing and restriction digest analysis, these shifts are caused by mono-allelic base transitions in exon 5 (c.547C-->T; R44X; We) and intron 12 (IVS12+5G-->A; Sc). Each mutation co segregates with the trait in the affected family with complete penetrance. The Sc mutation in the splicing donor site of intron 12 may result in either intron inclusion or exon skipping, both giving rise to a truncated PAX6 protein which may retain a residual transactivating activity. In contrast, the We genetic alteration is a loss-of-function mutation leading to a more severe phenotype than that observed in the Sc pedigree. PMID- 10694926 TI - A novel homozygous nonsense mutations E135* in the type II 3beta-hydroxysteroid dehydrogenase gene in a girl with salt-losing congenital adrenal hyperplasia. Mutations in brief no. 168. Online. AB - Mutations in the 3beta-hydroxysteroid dehydrogenase (3beta-HSD) type II gene have been reported in a small number of affected females. We report a 46,XX girl born to consanguineous parents from Chile. At birth, she had normal but hyperpigmented female external genitalia. At 60 days she presented salt loss. At 20 months, the diagnosis of classic salt-losing 3beta-HSD deficiency was made based on an elevated serum 17-hydroxpregnenolone concentration and a high 17 hydroxypregnenolone/17-hydroxyprogesterone ratio. Genomic DNA was amplified by PCR and screened for mutations by denaturing gradient gel electrophoresis and directly sequenced. A novel homozygous E135* mutation was found in the 3beta-HSD type II gene of the patient while her parents were heterozygotes. This novel nonsense homozygous E135* mutation led to encode a predicted truncated 134 amino acid protein instead of the native 371 amino acid 3beta-HSD type II protein. This predicted product is consistent with the severe 3beta-HSD deficiency in this girl. PMID- 10694927 TI - The distal boundary of myogenic primordia in chimeric avian limb buds and its relation to an accessible population of cartilage progenitor cells. AB - Using chimeras consisting of chick embryos that had received substitution grafts of quail somites, we have determined the distalmost extension of the myogenic primordia in the outgrowing wing bud at 5 days of incubation. At Hamburger Hamilton stage 25 the most distal premuscle cell is consistently 300 mum or more from the apex of the wing mesoblast. The stage 25 wing tip resembles very early whole limb buds in not having proceeded beyond the mesenchymal state or having expressed markers of terminal differentiation. However, unlike early whole limb buds it is free of a myogenic subpopulation. We therefore propose that the stage 25 wing tip is the appropriate system for in vitro and molecular studies of cartilage differentiation. PMID- 10694928 TI - Anti-neural antibodies in serum and cerebrospinal fluid of amyotrophic lateral sclerosis (ALS) patients. AB - OBJECTIVES: An autoimmune basis has been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). This hypothesis is supported by the presence of antibodies that interact with motoneuron antigens in serum of these patients. Against autoimmunity are the discrepances in the frequency of the antibodies appearance and also failure of immunosuppression. The aim of our study was to evaluate the titer of antibodies against GM1-gangliosides, AGM1-gangliosides and anti-sulfatides in paired serum and cerebrospinal fluid samples in the ALS patients. MATERIAL AND METHODS: Serum of 103 and CSF of 79 patients with ALS was examined. The "disease controls" consisted of 22 cases of other motor neuron diseases and 50 healthy, age-matched normals. CSF was drawn at the same time from 79 ALS patients, 6 cases of the "disease controls" and 50 normals. To study the titer of antibodies against GM1-gangliosides, AGM1-gangliosides and sulfatides the ELISA technique has been applied. RESULTS: An increased titer against GM1 gangliosides, AGM1-gangliosides and sulfatides in ALS appeared in serum in 18%, 32%, and 11%, resp., in the "disease controls" the increased antibodies titer appeared in single cases. In CSF the appropriate values in ALS were 20%, 15%, 8%, resp. In the "disease controls" a high antibodies titer was a rare finding. CONCLUSIONS: It is concluded that in some ALS cases and also in some patients with other motor neuron diseases an autoimmune mechanism may contribute to motor neuron injury. PMID- 10694929 TI - Evidence of 14C-furazolidone metabolite binding to the hepatic DNA of trout. AB - Furazolidone (FZ) is a nitrofuran drug commonly used in aquaculture. In the present study, [methylidene-14C]-FZ or [oxazolone-4,5-14C]-FZ was offered to rainbow trout (Oncorhyncus mykiss) in medicated feed at a daily dose of 135 mg/kg b. wt. for 10 days. The trout were sacrificed at specific time points post-dosing and the liver removed for DNA-bound 14C characterization. Both forms of the 14C labelled FZ were converted by trout to reactive metabolite(s) which bound irreversibly to the hepatic DNA. The amount of 14C bound to the hepatic DNA increased with post-dosing time and was higher in trout pretreated with [methylidene-14C]-FZ than in trout pretreated with [oxazolone-4,5-14C]-FZ. The identity of the FZ reactive metabolite(s) remained to be elucidated. However, a part of the FZ reactive metabolite(s) could be released as 3-amino-2-oxazolidone by acid hydrolysis. An appreciable amount of 14C was also found to bind irreversibly with the hepatic DNA of trout following an i.v. injection of [oxazolone-4,5-14C]-FZ. Results of these studies indicate that FZ is metabolized by trout to a reactive metabolite(s) which binds irreversibly to the DNA of trout liver. PMID- 10694930 TI - Induction and inhibition of cytochrome P450-catalysed reduction of biologically active benfluron N-oxide. AB - Benfluron N-oxide [5-(2-N-oxo-2-N,N'-dimethylaminoethoxy)-7-oxo-7-H benzo[c]fluorene] is a biologically active substance which displays a cytostatic effect on several experimental tumour cells. The main metabolic pathway of benfluron N-oxide in vitro and in vitro--its reduction to the parent tertiary amine benfluron--and the role of cytochrome P450 in this reduction were studied. The value of the benfluron N-oxide/benfluron redox potential as a criterion of suitability of the substrate for cytochrome P450 reductase activity was determined. Results of induction and inhibition studies on rats suggest that cytochromes P4502B and P4502E1 participate in microsomal reduction of benfluron N oxide. Unlike most cytochrome P450 catalysed reactions, the reduction of benfluron N-oxide also occurs under aerobic conditions. Microsomes induced by phenobarbital, ethanol or beta-naphthoflavone showed no significantly greater inhibitory effect of oxygen on benfluron N-oxide reduction. PMID- 10694931 TI - Inter-species comparison of microsomal reductive transformation of biologically active benfluron N-oxide. AB - Benfluron N-oxide is an anti-neoplastic active metabolite of benfluron (B) /1/. It is generated by flavine-monooxygenase-catalysed reactions /2/ and immediately undergoes subsequent metabolic transformations, the most important of which are reductive reactions /3/. The products of reductive pathways catalysed by two different microsomal enzymatic systems are the tertiary amine benfluron (i.e. the original parent compound) and/or 7-dihydrobenfluron N-oxide. Our studies on the reductive transformation of B N-oxide in rat, mouse, guinea-pig, rabbit, mini-pig and human microsomes have revealed significant species differences both in the yields of respective reduced metabolites and in the conditions essential for the activity of the reductases involved. While B, the original tertiary amine, is the main product of aerobic incubation of B N-oxide with NADPH in rat, mouse and mini pig, significantly higher activities of the enzymes catalysing the formation of 7 dihydro-B N-oxide have been detected in rabbit and human microsomes. In rat, mouse and mini-pig, NADPH rather than NADH is the preferred coenzyme for B formation, and NADPH is also the preferred coenzyme for the formation of 7 dihydro-B N-oxide in most of the species used. The yield of tertiary amine B is higher in anaerobic rather than aerobic conditions in most experimental species studied. Aerobic or anaerobic incubating conditions have an insignificant effect on the formation of 7-dihydro-B N-oxide. Based on the inhibitory effect of CO on the reductive transformation of B N-oxide, cytochromes P450 can be assumed to participate in the formation of B both in rat and mini-pig, and, in mini-pig only, also in the formation of 7-dihydro-B N-oxide. Inter-species comparison of the properties of the reductases participating in the transformation of B N-oxide shows that the rabbit is a suitable model to study reductive transformation of B N-oxide in man. PMID- 10694932 TI - Effects of carbamazepine on hepatic glutathione level in rats and determination of carbamazepine and its epoxide metabolite in plasma by HPLC. AB - We investigated whether carbamazepine, which is known to be metabolized to an electrophilic epoxide derivative in the body, causes any decrease, analogous to the action of epoxides, of hepatic glutathione (GSH) level in rats. Carbamazepine was administered to rats and liver GSH levels were determined spectrophotometrically. Neither a single low nor repeated low doses (30 mg/kg) of carbamazepine (CBZ) produced a statistically significant difference in GSH levels relative to controls. A single high dose of CBZ (100 mg/kg) produced a large and significant decrease relative to control (GSH level 3.82 +/- 0.64 vs 6.54 +/- 0.45 mumol GSH/g liver). CBZ and its metabolite carbamazepine-10,11-epoxide were determined in plasma by HPLC after the high dose of carbamazepine administration. The concentrations of carbamazepine and carbamazepine-10,11-epoxide were 18.9 +/- 2.9 micrograms/ml and 10.7 +/- 2.8 micrograms/ml, respectively. PMID- 10694933 TI - Tiagabine: absence of kinetic or dynamic interactions with ethanol. AB - Tiagabine is a new antiepileptic drug that inhibits the uptake of gamma aminobutyric acid into neurons and glia. This double-blind, placebo-controlled study investigated the effect of multiple doses of tiagabine on the adverse cognitive effects produced by a single dose of ethanol in 20 healthy volunteers. The effects of each drug on the pharmaco-kinetics of the other were also determined. Compared with placebo, tiagabine produced no statistically significant effects on digit vigilance speed (primary assessment variable) or accuracy, choice reaction time, immediate or delayed word recall, delayed word recognition speed or sensitivity, visual tracking, body sway, or subjective measures of alertness, calmness, and contentment. There was no evidence of a pharmacodynamic interaction between tiagabine and ethanol with respect to these variables. The pharmacokinetic parameters of tiagabine and ethanol (maximum plasma concentration [Cmax], time to Cmax and area under the plasma concentration time curve) were unchanged during concomitant administration. Adverse events, which mainly affected the central nervous system, occurred with a similar incidence during tiagabine and placebo administration and were more common after the administration of ethanol. There appears to be no need for additional caution regarding driving or operating machinery when ethanol is administered to patients taking tiagabine. PMID- 10694934 TI - Cadherin adhesion in the intestinal crypt regulates morphogenesis, mitogenesis, motogenesis, and metaplasia formation. AB - The topographical organisation of the epithelium lining mucous membranes has been an intense point of research. One of the fundamental biological issues underpinning this and associated issues relates to the role and regulation of epithelial adhesion molecules. Adhesion between individual cells allows an intact layer to be formed, which is selectively permeable. In addition, the orchestrated regulation of multiple adhesion molecules allows the gradual transition from basal secretory cells to apical absorptive cells in the crypt-villus gradient. Moreover, it is becoming clear that no one class of adhesion molecule can sufficiently govern crypt architecture; however, the main cell-cell adhesion molecules are the cadherins and the related desmosomal cadherins. These latter molecules interact with the catenins, which bind directly or indirectly with cytoskeletal molecules such as Rho and Rac. In addition, other complex glycoproteins, such as the carcinoembryonic antigens, might contribute to adhesion, although their mechanisms of function are distinctly different. Integrins on the basal aspect of the cells also signal important morphoregulatory signals as a result of their binding to the extracellular maxtrix. The disruption of these physiological processes also provides a necessary and, in some cases, sufficient molecular mechanism for cancer invasion and metastasis, such as occurs in E-cadherin mutation positive familial gastric cancer. PMID- 10694935 TI - The adenomatous polyposis coli protein. AB - Mutations in the adenomatous polyposis coli (APC) gene are associated with most colorectal cancers. The APC protein has been implicated in many aspects of tumour development. This article will discuss recent data suggesting that APC may have multiple functions in the cell. First, APC is a component of the Wnt signalling pathway; second, APC may have a role in cell migration; finally, APC may regulate proliferation and apoptosis. PMID- 10694936 TI - Lewis blood group and CEA related antigens; coexpressed cell-cell adhesion molecules with roles in the biological progression and dissemination of tumours. AB - The Lewis blood group and carcinoembryonic antigen (CEA) related antigens have adhesive functions in human tissues, with roles in embryonic sorting and migration of cells (organogenesis), differentiation and protection of normal mucosal tissues, migration of neutrophils, bacterial binding, and tumour differentiation and dissemination. In the key areas of mucosal protection, neutrophil binding, and tumour metastasis, they are often coexpressed on the outer cell membrane, with Lewis blood group antigens forming the terminal carbohydrate chains on a CEA related glycoprotein backbone. The central role of these antigens in the mechanism of neutrophil binding to endothelium in inflammation highlights a fascinating paradigm for tumour cell dissemination and metastasis, and expression is linked to disease prognosis. This review outlines the structure, function, and comparative roles of these antigens in human tissues. PMID- 10694937 TI - Regulation of E-cadherin: does hypoxia initiate the metastatic cascade? AB - The ability of tumours to metastasis is regarded as one of the hallmarks of malignancy. The process through which tumours evolve to achieve this has been termed the metastatic cascade. This cascade has been the subject of much investigation over many years. One of the vital events identified by these investigations is the reduction of adhesion between tumour cells facilitating invasion of the surrounding tissues and vascular channels, ultimately leading to the development of a distant metastasis. E-cadherin and its associated catenin complex have been identified as key molecules in cell adhesion. This review looks at the structure and interaction of the E-cadherin-catenin complex and the factors that appear to regulate E-cadherin expression and thus cell adhesion. From the data gathered, it has become possible to propose the hypothesis that the development of tumour hypoxia is the initiating factor that sets the tumour on the road to metastasis. PMID- 10694938 TI - CD44 cell adhesion molecules. AB - The CD44 proteins form a ubiquitously expressed family of cell surface adhesion molecules involved in cell-cell and cell-matrix interactions. The multiple protein isoforms are encoded by a single gene by alternative splicing and are further modified by a range of post-translational modifications. CD44 proteins are single chain molecules comprising an N-terminal extracellular domain, a membrane proximal region, a transmembrane domain, and a cytoplasmic tail. The CD44 gene has only been detected in higher organisms and the amino acid sequence of most of the molecule is highly conserved between mammalian species. The principal ligand of CD44 is hyaluronic acid, an integral component of the extracellular matrix. Other CD44 ligands include osteopontin, serglycin, collagens, fibronectin, and laminin. The major physiological role of CD44 is to maintain organ and tissue structure via cell-cell and cell-matrix adhesion, but certain variant isoforms can also mediate lymphocyte activation and homing, and the presentation of chemical factors and hormones. Increased interest has been directed at the characterisation of this molecule since it was observed that expression of multiple CD44 isoforms is greatly upregulated in neoplasia. CD44, particularly its variants, may be useful as a diagnostic or prognostic marker of malignancy and, in at least some human cancers, it may be a potential target for cancer therapy. This review describes the structure of the CD44 gene and discusses some of its roles in physiological and pathological processes. PMID- 10694939 TI - Small GTPases and regulation of cadherin dependent cell-cell adhesion. AB - Cadherins belong to a superfamily of cell-cell adhesion receptors that bind to the same type of molecules (homotypic interaction) in a calcium dependent manner. Different members of the family are found in a wide variety of cell types and cadherin adhesive function plays a role in cell fate, segregation, and differentiation, which ensures the higher order of organisation found in many tissues. This review will focus on the role that cadherin adhesiveness plays in the differentiation of epithelial cells, and how cadherin function can be regulated by proteins of the small GTPase family. In the text, readers are referred to recent reviews and other chapters covering important topics that are not discussed here because of space limitation. PMID- 10694940 TI - Alpha E beta 7. AB - alpha E beta 7 is a member of the integrin family and is expressed almost exclusively by cells of the T lymphocyte lineage in mucosal tissues. Expression is induced by transforming growth factor beta in the mucosal microenvironment. Genetic elements that control transcription are under investigation and may prove valuable for directing the expression of transgenes in mucosal T cells. The only known ligand for alpha E beta 7 is E-cadherin, which is expressed on epithelial cells. In this article, molecular aspects of ligand recognition by alpha E beta 7 in relation to recent structural data on cadherin domains are reviewed. Expression of alpha E beta 7 is often increased in inflammatory diseases, particularly where T cells infiltrate epithelial tissues. The function of alpha E beta 7 is not yet fully understood, but it is likely to be important in retention of T cells in mucosal tissues and may also have a role in cell signalling and communication between lymphocytes and epithelial surfaces. PMID- 10694941 TI - Integrins: a role as cell signalling molecules. AB - Integrins form the major family of proteins that mediates cell-matrix interactions. As well as an adhesive function, it is increasingly apparent that integrins can transduce messages via classic signalling pathways and impact upon such fundamental cellular processes as proliferation, apoptosis, differentiation, and motility. Dysregulation of these processes are a feature of many malignancies. Altered integrin expression has been observed in many human tumours, and perturbation of integrin function or expression in experimental systems has demonstrated that altered integrin signalling may directly contribute to the development of the malignant phenotype. PMID- 10694942 TI - Adhesion of lymphocytes to hepatic endothelium. AB - Chronic inflammation occurs when factors that regulate the process of leucocyte recruitment are disrupted, and it is dependent on recruitment, activation, and retention of lymphocytes within tissue microenvironments. The molecular mechanisms that mediate lymphocyte adhesion to vascular endothelial cells have been described by several groups, but the signals involved in the recruitment of lymphocytes via the hepatic circulation have yet to be elucidated fully. This article considers the liver as a model of organ specific lymphocyte recruitment. In this context, the roles of leucocyte and endothelial adhesion molecules and chemokines in lymphocyte recruitment are discussed. The article also reviews the mechanisms that regulate lymphocyte recirculation to the liver under both physiological and pathological conditions and draws parallels with other organs such as the gut and skin. PMID- 10694943 TI - Cell adhesion molecules in the pathogenesis of and host defence against microbial infection. AB - Eukaryotic cell adhesion molecules (CAMs) are used by various cells and extracellular molecules in host defence against infection. They are involved in many processes including recognition by circulating phagocytes of a site of inflammation, transmigration through the endothelial barrier, diapedesis through basement membrane and extracellular matrix, and release of effector mechanisms at the infected site. CAMs involved in leucocyte-endothelial cell interaction include the selectins, integrins, and members of the immunoglobulin superfamily. However, CAMs are also used by various microorganisms (protozoa, fungi, bacteria, and viruses) during their pathogenesis. For example, bacteria that utilise CAMs include Mycobacterium tuberculosis, Listeria monocytogenes, Yersinia spp, enteropathogenic Escherichia coli, Shigella spp, Neisseria spp, Bordetella spp, and Borrelia burgdorferi. In addition, CAMs are involved in the pathogenetic effects of the RTX toxins of Pasteurella haemolytica, Actinobacillus actinomycetemcomitans, and the superantigen exotoxins of Staphylococcus aureus and Streptococcus pyogenes. A recurrent and topical theme of potential importance within the bacterial group is the intimate relation between CAMs, bacterial protein receptors, and type III secretion systems. For example, the IpaBCD protein complex is secreted by the type III system of Shigella flexneri and interacts with alpha 5 beta 1 integrin on the eukaryotic cell surface, followed by Rho mediated internalisation; this illustrates the relevance of cellular microbiology. CAMs might prove to be novel therapeutic targets. Comparative genomics has provided the knowledge of shared virulence determinants among diverse bacterial genera, and will continue to deepen our understanding of microbial pathogenesis, particularly in the context of the interaction of prokaryotic and eukaryotic molecules. PMID- 10694944 TI - A colorectal cell line with alterations in E-cadherin and epithelial biology may be an in vitro model of colitis. AB - BACKGROUND: It has been shown previously in ulcerative colitis tissue that E cadherin can occasionally be mutated in the extracellular domain early in neoplastic progression. E-cadherin is known to maintain differentiation and inhibits invasion in vivo. AIMS: To assess the mechanisms by which such dysfunction occurs. METHODS: Four human colorectal cancer cell lines, HCA-7 colonies 1, 3, 6, and 30, derived from a single heterogeneous colorectal cancer were studied. The HCA-7 cell line has p53 mutations and a random errors of replication "positive" phenotype, as is seen in early colitis associated cancers or hereditary nonpolyposis coli cancer (HNPCC). RESULTS: Cell lines 6 and 30 expressed E-cadherin abundantly and this correlated positively with their degree of differentiation and organisation; however, both cell lines had loss of heterozygosity of E-cadherin. Interestingly, E-cadherin production was downregulated in the poorly differentiated cell line 1, and this was associated with major chromosomal rearrangements of 16q. This cell line also had a mutation in the homophilic binding domain of exon 4, which was associated with disaggregation by low titres of a function blocking antibody, and an invasive phenotype. CONCLUSIONS: These multiple biological alterations further characterise the complex association that E-cadherin has with tumour heterogeneity and suggest that this series of cell lines may be a useful model of colitis associated or HNPCC associated tumorigenesis. PMID- 10694945 TI - Prevention and treatment of chemotherapy- and radiotherapy-induced oral mucositis: a review. AB - Oral mucositis is a distressing toxic effect of systemic chemotherapy with many commonly utilized drugs and of head and neck irradiation in patients with cancer. The agents and methods that have been used and studied in chemotherapy- and radiotherapy-induced oral mucositis, their mechanisms of action, and the current knowledge of their efficiency to reduce the incidence, severity or shorten the duration of oral mucositis are reviewed in this article. Oral cooling is a cheap and available method to lower the severity of bolus 5-fluorouracil-induced oral mucositis. However, more effective methods are needed. Results of studies with granulocyte-macrophage colony-stimulating factor or granulocyte colony stimulating factor are promising. Lasers are partly beneficial, but equipment demanding. Modification of the chemotherapy regimen resulting in shortening of the exposition time to chemotherapy agents or chronomodulation of chemotherapy has been shown to lower mucosal toxicity of some regimens. Results of animal studies with locally applied transforming growth factor beta 3 and interleukin-11 are also promising. Based on the findings of the role of the inflammatory cascade in the response of normal tissues to chemotherapy and radiotherapy, anti inflammatory drugs might be beneficial. At the present time, no agent has been shown to be uniformly efficacious and can be accepted as standard therapy of chemotherapy- and radiotherapy-induced oral mucositis. Further intensive research is needed. PMID- 10694946 TI - Cancer of mouth, pharynx and nasopharynx in Asian and Chinese immigrants resident in Thames regions. AB - Studies on migrants can generate important clues on the etiology of cancer. The purpose of the present study was to determine the relationship between ethnic origin and the incidence of oral and pharyngeal cancers among residents of the Thames regions in southern England. Records from the Thames Cancer Registry during the period 1986-91 were examined and south Asians and Chinese ethnic immigrants flagged using their place of birth and names. Computation of relative incidence among head and neck cancers (n = 7222) showed that oral cancer was significantly higher among Asians (95/232 = 40.9%) and nasopharyngeal cancer among Chinese (45/67 = 67.2%). Some differences in the intra-oral site of cancer and ethnic origin were noted. The ethnic migrants were significantly younger (Asians 51.6 +/- 34.8 years, Chinese 47.6 +/- 14.8 years) compared to the rest of the population (64.8 +/- 15.6 years) at the time of cancer diagnosis (p = 0.0) but no significant differences were found for the stage of presentation. The mean survival period for a cancer of the head and neck was 2.2 years and significant differences in cumulative rates of survival were noted among the three groups studied (p = 0.003). A strong correlation was noted between the incidence of oral cancer and local authorities with a high percentage of Asian residents. The south Asian and Chinese ethnic minorities constitute important high risk groups for oral and nasopharyngeal cancer, for whom targeted prevention is indicated. PMID- 10694947 TI - Inactivation patterns of the p16 (INK4a) gene in oral squamous cell carcinoma cell lines. AB - To determine whether inactivation of the p16 gene mapped to the chromosome 9p21 region is associated with the development of oral squamous cell carcinoma (SCC), we investigated the mutational states of two forms of alternative transcripts (alpha and beta) from the p16 gene in 14 oral SCC cell lines by means of RT-PCR, PCR, direct sequencing and methylation analyses. Alterations of the alpha transcript were detected in all of the cell lines examined: homozygous deletions in three lines; subtle mutations in exons 1 alpha or 2 in four lines; skipping of exon 2 in two lines; hypermethylation of the 5' CpG island of the p16 gene in four lines; and an unknown mechanism in one line. On the other hand, abnormalities of the beta transcript were observed in seven of the 14 cell lines. Nonetheless, the mutations that essentially affect the function of the encoded protein were found only in five cell lines, including three lines with homozygous deletion. There was no cell line having only beta transcript alterations. Thus, alteration of the alpha transcript of the p16 gene was a highly frequent event in oral SCC. Since this type of alteration resulted in gene inactivation through multiple pathways, it may play a major role in the process of oral SCC development. PMID- 10694948 TI - Prognostic importance of the expression of CD44 splice variants in oral squamous cell carcinomas. AB - Considering squamous cell carcinomas (SCCs) of the oral cavity and oropharynx the molecular mechanisms underlying the infiltration and destruction of adjacent tissue as well as the metastatic spread are largely unknown. In this context, the detection of defective expression of cellular adhesion molecules in the tumour cells, e.g. CD44, might be important and correlated with prognosis. Paraffin embedded tumour-tissue from 99 patients with primary oral and oropharyngeal SCC, additionally including corresponding lymph-node metastases in nine cases, was analysed for expression of the CD44 splice variants v4, v5, v6, v7, and v9 by means of immunohistochemistry. A diminution of at least one of the examined CD44 isoforms compared to the normal oral epithelium was observed in 39.4% of the squamous cell carcinomas. No correlations could be found between CD44 expression and pT- or pN-stage. However, decreased expression of v9 was correlated with higher histological grade (p < 0.001). Moreover, reduced CD44 expression was a statistically significant independent predictor for shorter survival time (p = 0.002) as well as shorter recurrence-free interval (p = 0.004) in addition to pT- and pN-stage. The separate analysis showed that particularly the decreased v7 (p = 0.04) and v9 (p < 0.02) expression in the tumour cells was associated negatively with survival. PMID- 10694949 TI - Carbon dioxide laser evaporation of leukoplakia of the lower lip: a retrospective evaluation. AB - The purpose of this study was the retrospective evaluation of the treatment results of CO2 laser evaporation for 27 cases of leukoplakia of the lip. The data were derived from 23 patients who presented with leukoplakia of the lower lip during the period 1978-96. Four patients developed a second primary leukoplakia of the lip resulting in 27 cases of leukoplakia. All lesions were treated with a CO2 laser equipped with an operation microscope and micromanipulator. Short-term evaluation showed complete epithelialisation 4 weeks after CO2 laser evaporation; there was minimal scar formation and no subsequent interference with normal lip function. During long-term evaluation, four recurrences (14.8%) were diagnosed which developed between 5 and 31 months after treatment, these were retreated with CO2 laser evaporation. There was no development of squamous cell carcinoma in the CO2 laser-treated area. Selective removal of affected epithelium with minimal damage to surrounding structures is possible using CO2 laser evaporation, followed by excellent wound healing and good functional result. Treatment can be performed under local anaesthesia on an outpatient basis. The recurrence rate is low compared with the recurrence rate after surgical excision. Therefore, CO2 laser evaporation is considered a reliable and effective treatment modality for leukoplakia of the lip. PMID- 10694950 TI - Eosinophil ablation and tumor development. AB - Tissue eosinophilia in squamous cell carcinoma has long been recognized; however, the role of eosinophils in tumor development remains unclear. Studies have reported both favorable and unfavorable prognoses for patients with tumors exhibiting tumor-associated tissue eosinophilia (TATE). This study seeks to elucidate the potential role of the eosinophil in squamous cell carcinoma development and provide an experimental model for future studies. The carcinogen induced hamster oral cancer model was found to fulfill these objectives. Eosinophils progressively infiltrate into this carcinogen-induced oral cancer model. We now demonstrate that TATE is completely abolished by the use of an anti interleukin-5 monoclonal antibody (mAb) preparation, TRFK-5. Clinical observations revealed that TRFK-5-treated hamsters exhibited smaller tumor burden and delayed onset of tumor development. The results suggest that anti-interleukin 5 antibody treatment may delay and/or inhibit tumor development, and that eosinophils may have a tumor-promoting role. PMID- 10694951 TI - Prognostic significance of proliferative and apoptotic markers in oral tongue squamous cell carcinomas. AB - The prognostic impact of proliferative and apoptotic markers was studied in 85 T1 4 oral tongue squamous cell carcinomas (SCCs). Ki67 immunoreactivity and AgNOR counts, including mean AgNOR counts (mAgNOR) and the percentage of nuclei with more than one AgNOR (pAgNOR > 1), were used as proliferative parameters. The apoptotic index (AI) was assessed using the TUNEL method. Bax expression was detected immunohistochemically and scored. Bax expression correlated positively with AI (p = 0.0122). Ki67 correlated with both pAgNOR > 1 (p = 0.0042) and mAgNOR (p = 0.0189). Low Bax expression and low AI correlated significantly with the disease-free period (p = 0.0001 and p = 0.0024, respectively). High values for Ki67, pAgNOR > 1 and mAgNOR correlated with poor prognosis (p = 0.0021, p = 0.0001 and p = 0.0244, respectively). Combinations of proliferative and apoptotic parameters were stronger predictors than individual parameters (p < 0.0001). pAgNOR > 1-Bax expression appeared to be the best combination (p < 0.0001). We conclude that proliferative and apoptotic markers, especially their combinations, have prognostic value in tongue SCC. PMID- 10694952 TI - Evaluation of telomerase activation in head and neck cancer. AB - During replication of the linear chromosomes, telomeres, i.e. the ends of the chromosomes, are not replicated completely by the conventional DNA polymerases. Therefore, normal somatic cells senesce after certain number of cell divisions. Telomerase is a special reverse transcriptase used by most eukaryotes to achieve immortalization. Telomerase activity has been determined in a variety of cancers. However, there are few reports on telomerase activity in head and neck cancer. The etiology of the disease in India is completely different from Western countries. Tobacco consumption is more prevalent in India and the mode of tobacco consumption (e.g. chewing, snuffing, bidi smoking, reverse smoking) is also different. The present study determined telomerase activity in 32 malignant tumour samples of head and neck cancer patients, 11 samples from patients with precancerous/benign lesions and 30 samples of adjacent normal tissues. Telomerase was found to be activated in 80% of the patients with head and neck cancer, 100% of the patients with precancerous/benign lesions and 74% of the adjacent normal tissues. According to the theory of field cancerization, carcinogenic insults (e.g. tobacco) may result into multiple malignant foci. This fact may explain the reason for high telomerase positivity in adjacent normal as well as precancerous/benign tissues. Telomerase activation and the clinical or histopathological characteristics of the head and neck cancer patients were observed to be independent features. This is a preliminary report which has generated a greater interest for in-depth elucidation of the role of telomerase and telomeres in head and neck carcinogenesis in India. PMID- 10694953 TI - Differential DNA methylation of the p16 INK4A/CDKN2A promoter in human oral cancer cells and normal human oral keratinocytes. AB - The p16 INK4A tumor suppressor gene participates in establishing and maintaining the malignant phenotype of a variety of cancer cell lines and primary tumors. Recently it has been observed that p16 expression is lost in oral cavity cancer cell lines in the presence of a normal intact gene. To examine the role of DNA methylation as an explanation for these findings, we analyzed the DNA methylation patterns of the p16 INK4A promoter in DNA isolated from primary cultures of normal human oral keratinocytes and squamous cell carcinoma (SCC-15) oral cancer cells using bisulfite genomic sequencing. Our results demonstrated striking differences in the methylation status of the 5' CpG island of the p16 gene between normal and cancer cells. Normal human oral keratinocytes showed practically no methylation of the p16 INK4A promoter, while SCC-15 oral cancer cells showed almost complete methylation in this region. These data implicate DNA methylation as a mechanism for transcriptional silencing of the p16 INK4A gene in oral cancer cells. PMID- 10694954 TI - Effects of bisphosphonate on experimental jaw metastasis model in nude mice. AB - The mechanism of osteolysis associated with metastatic cancer of the jaws is essentially osteoclast-mediated. Therefore, it is likely that potent osteoclastic bone resorption inhibitors such as bisphosphonates would be efficacious for the treatment of jaw metastasis. We examined the effects of a third generation bisphosphonate, YM175, in a nude mice jaw metastasis model with intracardiac injection of a human breast cancer cell line, MDA-MB-231. The metastatic lesions in untreated mice were radiographically observed at the body and angle of the mandible. Histology of the mandible of untreated mice revealed that most of the bone marrow cavities had been occupied by the metastatic tumor with active osteoclasts along the trabecular bone. The experimental group showed that YM175 markedly reduced the size of tumor and the number of osteoclasts. These results suggest that YM175 may suppress metastasis formation and tumor growth in jaw through inhibition of osteoclastic bone resorption. PMID- 10694955 TI - Well-differentiated intraosseous osteosarcoma of the jaws: experience of two cases from the Instituto Nacional de Cancerologia, Mexico. AB - Osteosarcomas of the jaws represent less than 10% of all osteosarcomas, and most of them are high-grade neoplasms. Prognostic factors in overall survival include tumor size, location and histologic grade. Examples of well-differentiated (low grade) intraosseous osteosarcomas of the jaws (WDIOJ) have been rarely reported. This article presents two cases of this unusual lesion, one of which was located in the maxilla of a 17-year-old man and the other developed in the mandible of a 37-year-old woman. CT scan was necessary to detect the small foci of penetration into the thinned cortical bone and the reactive periosteal bone formation, which are important findings to establish the correct diagnosis of WDIOJ and help to exclude other benign intraosseous lesions that may be very similar histologically, such as fibrous dysplasia, ossifying and desmoplastic fibromas. In spite of tumor size (mean 5.2 cm), their well-demarcated borders allowed complete removal of both tumors. There is no evidence of tumoral activity in any of our patients after follow-up periods of 15 months and 5 years. Wide excision seems to be the treatment of choice for this subgroup of osteosarcomas. PMID- 10694956 TI - The rise and fall of a theory of vision--Hecht's photochemical doctrine. PMID- 10694957 TI - Gorillas in our midst: sustained inattentional blindness for dynamic events. AB - With each eye fixation, we experience a richly detailed visual world. Yet recent work on visual integration and change direction reveals that we are surprisingly unaware of the details of our environment from one view to the next: we often do not detect large changes to objects and scenes ('change blindness'). Furthermore, without attention, we may not even perceive objects ('inattentional blindness'). Taken together, these findings suggest that we perceive and remember only those objects and details that receive focused attention. In this paper, we briefly review and discuss evidence for these cognitive forms of 'blindness'. We then present a new study that builds on classic studies of divided visual attention to examine inattentional blindness for complex objects and events in dynamic scenes. Our results suggest that the likelihood of noticing an unexpected object depends on the similarity of that object to other objects in the display and on how difficult the priming monitoring task is. Interestingly, spatial proximity of the critical unattended object to attended locations does not appear to affect detection, suggesting that observers attend to objects and events, not spatial positions. We discuss the implications of these results for visual representations and awareness of our visual environment. PMID- 10694958 TI - Driving experience and the functional field of view. AB - Research has suggested that novice drivers have different search strategies compared with their more experienced counterparts, and that this may contribute to their increased accident liability. One issue of concern is whether experienced drivers have a wider field of peripheral vision than less experienced drivers. This study attempted to distinguish between people of varying driving experience on the basis of their functional fields of view. Participants searched video clips taken from a moving driver's perspective for potential hazards while responding to peripheral target lights. Hit rates for peripheral targets decreased for all participant groups as processing demands increased (i.e. when hazards occurred) and as the eccentricity of the target increased, though there was no interaction. An effect of experience was also found which suggests that this paradigm measures a perceptual skill or strategy that develops with driving experience. PMID- 10694959 TI - Temporal constraints on visual learning: a computational model. AB - Given a constant stream of perceptual stimuli, how can the underlying invariances associated with a given input be learned? One approach consists of using generic truths about the spatiotemporal structure of the physical world as constraints on the types of quantities learned. The learning methodology employed here embodies one such truth: that perceptually salient properties (such as stereo disparity) tend to vary smoothly over time. Unfortunately, the units of an artificial neural network tend to encode superficial image properties, such as individual grey level pixel values, which vary rapidly over time. However, if the states of units are constrained to vary slowly, then the network is forced to learn a smoothly varying function of the training data. We implemented this temporal-smoothness constraint in a backpropagation network which learned stereo disparity from random-dot stereograms. Temporal smoothness was formalized with the use of regularization theory by modifying the standard cost function minimised during training of a network. Temporal smoothness was found to be similar to other techniques for improving generalisation, such as early stopping and weight decay. However, in contrast to these, the theoretical underpinnings of temporal smoothing are intimately related to fundamental characteristics of the physical world. Results are discussed in terms of regularization theory and the physically realistic assumptions upon which temporal smoothing is based. PMID- 10694960 TI - Do 9-month-olds perceive causation-at-a-distance? AB - Humans understand mechanical events to involve physical bodies interacting by contact, but intentional events involve agents that can also interact at a distance. We investigated infant sensitivity to causality in a simple event in which one agent appears to react to another without contact. Infants 9 months old were habituated to one of two events involving a computer-animated red square moving nonrigidly--like a caterpillar--towards a green square. In the 'reaction event', the green object moved in turn before the red one stopped, while in the 'pause event' the green object moved after the red one stopped. After habituation, each infant saw the habituation movie played in reverse. This test involved identical spatiotemporal changes for reaction and pause event, but the reversed reaction additionally involved a change in the causal roles. Infants dishabituated to reversal of the reaction but not the pause event, a result which suggests sensitivity to causation-at-a-distance. This ability could support development of social cognition and theory of mind. PMID- 10694961 TI - Fooling the eyes: trompe l'oeil and reverse perspective. AB - Trompe l'oeil pictures have been produced for hundreds of years. They attempt to create the impression of a surface that has different three-dimensional structure to the work; successful examples of trompe l'oeil typically constrain the observer's viewpoint and require use of a single eye. The works of Patrick Hughes are in relief but are painted to appear like conventional flat pictures; those parts that protrude from the picture plane are pictorially distant, or in reverse perspective. Movements of the observer result in fluid distortions of the pictorial image. These distortions occur with binocular observation and over a wide range of viewing distances. PMID- 10694962 TI - An analysis of binocular slant contrast. AB - When a small frontoparallel surface (a test strip) is surrounded by a larger slanted surface (an inducer), the test strip is perceived as slanted in the direction opposite to the inducer. This has been called the depth-contrast effect, but we call it the slant-contrast effect. In nearly all demonstrations of this effect, the inducer's slant is specified by stereoscopic signals; and other signals, such as the texture gradient, specify that it is frontoparallel. We present a theory of slant estimation that determines surface slant via linear combination of various slant estimators; the weight of each estimator is proportional to its reliability. The theory explains slant contrast because the absolute slant of the inducer and the relative slant between test strip and inducer are both estimated with greater reliability than the absolute slant of the test strip. The theory predicts that slant contrast will be eliminated if the signals specifying the inducer's slant are consistent with one another. It also predicts reversed slant contrast if the inducer's slant is specified by nonstereoscopic signals rather than by stereo signals. These predictions were tested and confirmed in three experiments. The first showed that slant contrast is greatly reduced when the stereo-specified and nonstereo-specified slants of the inducer are made consistent with one another. The second showed that slant contrast is eliminated altogether when the stimulus consists of real planes rather than images on a display screen. The third showed that slant contrast is reversed when the nonstereo-specified slant of the inducer varies and the stereo specified slant is zero. We conclude that slant contrast is a byproduct of the visual system's reconciliation of conflicting information while it attempts to determine surface slant. PMID- 10694963 TI - Test of Petter's rule for perceived surface stratification. AB - Petter's rule applies to two-dimensional patterns formed by two overlapping surfaces that alternatively appear in front of one another. It states that the surface with the shorter contours in the region where the surfaces look superimposed has a greater probability of appearing in front of the other surface. An experiment is reported the results of which show that Petter's rule is valid for chromatically homogeneous and for uniformly dense dotted patterns, and invalid for different kinds of chromatically inhomogeneous patterns. Petter's rule has been found to be valid when the overlapping surfaces have contours with gaps. It is proposed that Petter's rule derives from the dynamics of filling-in of contour gaps. PMID- 10694964 TI - Aftereffects and the representation of stereoscopic surfaces. AB - The structure of human disparity representation is examined through (i) adaptation experiments and (ii) model simulations of the data. Section 3 presents results of adaptation experiments designed to illuminate the structure of human disparity representation. Section 4 presents model simulations of three different disparity representation schemes. In the experiments, participants adapted to a 0.133 cycle deg-1 sinusoidally corrugated surface with 10 min of arc peak-to trough disparity. A flat test surface was briefly presented, in which the aftereffect surface was perceived. Adapt and test surfaces were placed on disparity pedestals and thus presented in front of or behind the plane of fixation. The adapt surface could be offset from the fixation plane by +/- 8 to 24 min of arc. The test surface could be offset from the fixation plane by +/- 8 to 48 min of arc. The depth aftereffect was measured in different disparity planes by a nulling method and 'topping-up' procedure. Aftereffect tuning functions were obtained whose bandwidths, magnitudes, and tuning depended on the disparity planes of both the adapt and test surfaces. These parameters were used to constrain the models tested in section 4. On the basis of the two studies, it is argued that the human stereoscopic system encodes spatial changes of disparity using channels localised within disparity planes. A localised disparity-gradient model of the human representation of disparity is proposed. PMID- 10694965 TI - Matching person identity from facial line drawings. AB - The processing of facial line drawings was investigated in either simultaneous or sequential matching trials with either the same or different viewpoint, showing pictures of faces either in the same modes (both photographs or line drawings) or different modes (one in each mode). Line drawings were particularly difficult to match in memory rather than under perceptual conditions, and line drawings did not allow the creation of efficient structural codes. These deficits of line representations underline the assumption that the face-processing system is inflexible when it is confronted with edge-based material. PMID- 10694966 TI - Brain scanning and the single mind. PMID- 10694967 TI - Differences in top-down influences on the reversal rate of different categories of reversible figures. AB - Understanding the mechanisms underlying the multistability of reversible figures may provide valuable insights into the normal functioning of our visual system. The proposed factors that control the perceptual alternations of reversible figures can be classified into bottom-up and top-down processes. In the present study, we report differences in top-down effects on the reversal rate depending on whether a structural perspective (Necker cube, Schroder staircase) or a meaningful content (duck/rabbit figure, chef/dog figure) is subject to the reversal phenomenon. In order to activate top-down mechanisms explicitly the subjects had the instruction to bring the reversal rate under voluntary control. The results indicated that both slowing down and speeding up the rate of alternations was more effective for the content-reversal figures (duck/rabbit, chef/dog) than for the rather abstract perspective-reversal figures (Necker cube, Schroder staircase). In order to investigate the effect of meaningfulness in figure/ground reversals, the effect of the same instructional variable was also determined for Rubin's vase/faces and the Maltese cross. The results showed a similar tendency as in the case of the comparison between perspective reversals and content reversals. Possible cognitive processes that may play a role in top down influences on figure reversal and theoretical implications of these findings for the interaction of bottom-up and top-down processes are discussed. PMID- 10694968 TI - Recognizing silhouettes and shaded images across depth rotation. AB - Outline-shape information may be particularly important in the recognition of depth-rotated objects because it provides a coarse shape description which gives first-pass information about the structure of an object. In four experiments, we compared recognition of silhouettes (showing only outline shape) with recognition of fully shaded images of objects, by means of a sequential-matching task. In experiments 1 and 2, the first stimulus was always a shaded image, and the second stimulus was either a shaded image or a silhouette. Recognition costs associated with a change in viewpoint were no greater for silhouettes than they were for shaded images. Experiments 3 and 4 replicated the design of the earlier experiments, but showed a silhouette as the initial stimulus, rather than a shaded image. In these cases, recognition costs associated with a change in viewpoint were greater for silhouettes than for shaded images. Combined, these results indicate that, while visual representations clearly include additional information, outline shape plays an important role in object recognition across depth rotation. PMID- 10694969 TI - Effects of face configuration change on shape perception: a new illusion. AB - We report two experiments indicating that varying the configuration of face features changes perception of an oval aperture windowing the face: as the eyes and mouth of a frontal-view face photograph are moved vertically toward face boundaries, the oval appears increasingly clongated, taller, and narrower; when eyes and mouth are moved toward the nose, the oval appears increasingly rounder, shorter, and wider. This shape illusion is maximised when faces appear upright within the oval and major face features (eyes, nose, and mouth) appear in their correct relative locations. These results establish that processing of a face configuration can affect perception of a geometric shape that shares visual space with a face. Whether the illusion is face-specific or a special case of a more general geometric illusion is discussed. PMID- 10694970 TI - Thompson's Margaret Thatcher illusion: when inversion fails. AB - It is argued that the whole face is more dominant than the individual features. In the case of a jumbled face the external pattern is dominant when a face is upright, whereas the internal pattern is dominant when a face is inverted. PMID- 10694971 TI - Wakes and spokes: new motion-induced brightness illusions. AB - Under certain conditions, high-contrast moving figures induce adjacent illusory regions, 'wakes' and 'spokes', which have contrast polarity opposite the inducing figures. In this paper we document properties of these novel phenomena. When the illusions are induced by a moving bar, spokes appear on the side of the bar closer to fixation and connect the bar to the fixation point, regardless of the momentary position of the bar or whether it is moving to the left or to the right. Although spokes often extend up to the fixation point, they never extend beyond it. This is not due to blocking of the spoke's spread by the fixation point, because in another experiment spokes extend directly through an intervening figure. Whereas spokes emanate from the end of a horizontally moving bar closest to fixation, wakes emanate from the end farthest from fixation. In contrast to spokes, wakes do not show a towards-fixation bias. Instead, the wake's end trails the position of the bar, like a ship's wake. The higher the bar velocity, the more the end of the wake appears to trail it, suggesting that wakes are caused by a process which spreads from the edge of moving figures. Wakes and spokes, as distinct illusions, should provide significant constraints on theories of human motion and brightness perception processes. PMID- 10694972 TI - Motion parallel to line orientation: disambiguation of motion percepts. AB - Four experiments demonstrate that lines indicating path of movement can generate rotational percepts in a multistable motion display that usually produces only horizontal or vertical motion percepts. The properties of the path-of-movement lines are predicted by a neural-network theory of visual perception. Experimental results validate the theory's predictions by demonstrating that movement of the display elements seems to follow an increasing luminance gradient in lines but not bars, and that illusory contours have similar effects. Experimental results also demonstrate that, in a choice between movement along lines drawn parallel or orthogonal to possible motion paths, observers more often see movement along the lines parallel to the motion path. These results suggest modifications to current computational and neurophysiological theories of motion perception. PMID- 10694973 TI - Do monocular time-to-collision estimates necessarily involve perceived distance? AB - Motivated by the debate between indirect and direct theories of perception, a large number of researchers have attempted to determine whether judgments of time to collision are based on the ratio of perceived distance to perceived speed or on the ratio theta/(d theta/dt), i.e. tau. Despite the considerable research effort devoted to this question there seems to be no clear resolution. We used a staircase tracking procedure to estimate errors in estimating time to collision for a simulated approaching object. To investigate the role of perceived distance in the judgment of time to collision, we asked observers to alternate between two viewing distances (100 and 500 cm). For the 500 cm viewing distance, we magnified the visual display by a factor of five so that the retinal images [and the values of theta/(d theta/dt) through time] were identical for the two viewing distances. All visual cues to distance were available. There were no significant differences between estimates of time to collision made at the two viewing distances. We conclude that our observers ignored perceived distance when estimating time to collision and based their responses on theta/(d theta/dt). We concur with recent proposals that, in the future, time-to-collision research should move away from the either/or analysis of different information sources that has dominated previous studies towards investigations of how different information sources are integrated. PMID- 10694974 TI - Exocentric pointing in three-dimensional space. AB - A study is reported of an exocentric pointing task in all three dimensions, in near space, with only two visible luminous objects--a pointer and a target. The task of the subject was to aim a pointer at a target. The results clearly show that visual space is not isotropic, since every set direction appeared to consist of two independent components--one in the projection onto a frontoparallel plane (tilt), the other in depth (slant). The tilt component shows a general trend across subjects, an oblique effect, and can be judged monocularly. The slant component is symmetrical in the mid-sagittal plane, requires the use of binocular information, and shows considerable differences between subjects. These differences seem to depend on the amount of binocular information used by each subject. There was a remarkably high level of consistency in the exocentric pointing, despite the absence of environmental cues. The within-subject consistency in the settings of the pointer corresponds to a consistency of about 1 min of arc in disparity of its tip, even though the pointer and target are separated by more than 5 deg. PMID- 10694975 TI - Effects of similarity in bandwidth on the auditory sequential streaming of two tone complexes. AB - We investigated the perceptual grouping of sequentially presented sounds- auditory stream segregation. It is well established that sounds heard as more similar in quality, or timbre, are more likely to be grouped into the same auditory stream. However, it is often unclear exactly what acoustic factors determine timbre. In this study, we presented various sequences of simple sounds, each comprising two frequency components (two-tone complexes), and measured their perceptual grouping. We varied only one parameter between trials, the intercomponent separation for some of the complexes, and examined the effects on stream segregation. Four hypotheses are presented that might predict the extent of streaming. Specifically, least streaming might be expected when the sounds were most similar in either (1) the frequency regions in which they have energy (maximum spectral overlap), (2) their auditory bandwidths, (3) their relative bandwidths, or (4) the rate at which the two components beat together (intermodulation rate). It was found that least streaming occurred when sounds were most similar in either their auditory or their relative bandwidths. Although these two hypotheses could not be distinguished, the results were clearly different from those predicted by hypotheses (1) and (4). The implications for models of stream segregation are discussed. PMID- 10694976 TI - A comment on "assimilation of achromatic color cannot explain the brightness effects in the achromatic neon effect" by Marc K Albert. PMID- 10694977 TI - Analysis of the proteome of Mycobacterium tuberculosis in silico. AB - Novel bioinformatics routines have been used to provide a more detailed definition of the proteome of Mycobacterium tuberculosis H37Rv. Over half of the current proteins result from gene duplication or domain shuffling events while one-sixth show no similarity to polypeptides described in other organisms. Prominent among the genes that appear to have been duplicated on numerous occasions are those involved in fatty acid metabolism, regulation of gene expression, and the unusually glycine-rich PE and PPE proteins. Protein similarity analysis, coupled with inspection of the genetic neighbourhood, was used to explore possible functional relatedness. This uncovered four large mce operons whose proteins may mediate initial interactions between the tubercle bacillus and host cells, together with a cluster of genes that might encode components of a structure required for secretion of ESAT-6 like proteins. Close linkage of the mmpL genes, encoding large membrane proteins, with those required for fatty acid metabolism suggests involvement in lipid transport. Compared to free-living bacteria, M. tuberculosis has a significantly smaller transport protein repertoire and this may reflect its intracellular lifestyle. PMID- 10694978 TI - Detection of mutations in drug resistance genes of Mycobacterium tuberculosis by a dot-blot hybridization strategy. AB - SETTING: Mycobacterium tuberculosis isolates from patients in communities endemic for tuberculosis in South Africa. OBJECTIVE: To develop a reliable PCR-based dot blot hybridization strategy to detect mutations conferring drug resistance. DESIGN: Different loci in six genes associated with drug resistance to isoniazid, rifampacin, streptomycin and ethambutol were selected to develop the PCR-based dot-blot hybridization strategy. RESULTS: Primers and probes to detect mutations at codons 315, 463 (katG) 269 (kasA), 531, 526 (rpoB) 43 (rpsL), 513 (rrs) and 306 (embB) were designed and used to develop a PCR-based dot-blot hybridization strategy. The dot-blot hybridization strategy with wild-type probes can efficiently be used to detect drug resistant mutations since these do not hybridize to mutant loci. Stripped blots and mutant probes can be used to identify the precise mutation. The embB gene (ethambutol resistance) was used to show how the dot-blot strategy can assist with the prediction of drug resistance more accurately. The method is rapid, reproducible, not technically demanding and samples can be done in batches. Additional loci can easily be incorporated. CONCLUSIONS: A PCR-based dot-blot hybridization strategy is described which can accurately identify drug resistant strains and the method is useful for patients at risk and in areas endemic for tuberculosis. PMID- 10694979 TI - Disruption of coding regions by IS6110 insertion in Mycobacterium tuberculosis. AB - SETTING: The insertion sequence IS6110 is widely used as a DNA fingerprinting probe for the classification of Mycobacterium tuberculosis strains. This study has focused on the characterization of regions disrupted by insertion of the IS6110 element. OBJECTIVE: To characterize IS6110 insertion loci in clinical isolates of M. tuberculosis, in terms of their genomic location and genetic identity, to ascertain whether IS6110 transposition could be a mechanism driving phenotypic change. DESIGN: Thirty-three IS6110 insertion loci were cloned from 8 clinical isolates of M. tuberculosis. Clones representing DR locus insertions were identified by hybridization (n = 4), and all other clones were characterized by DNA sequencing (n = 29). The sequence data was analyzed in conjunction with that of 43 other insertion loci identified in published literature and DNA sequence databases. RESULTS: The 76 sequences analyzed represented 66 unique insertion loci (including 9 unique insertions into the ipl locus). When mapped to the H37Rv genome, the majority of unique insertion loci demonstrated disruption of coding regions by IS6110 (n = 42; including the ipl insertions), while the remainder either occurred within intergenic regions (n = 17), or could not be mapped to the H37Rv genome sequence (n = 7). Mapping of the insertion loci reveals distribution throughout the chromosome, with isolated preferential insertion loci. CONCLUSIONS: This study has demonstrated the occurrence of 66 unique IS6110 insertion loci dispersed throughout the M. tuberculosis genome, with an unexpectedly high incidence of IS6110 insertions occurring within coding regions. However, the IS6110-mediated coding region disruptions identified here may only have limited impact on phenotype, as most of the coding regions disrupted are members of multiple gene families. Disruption of individual members of a family of genes may have no effect on phenotype or could have a minor or major impact, depending on the specificity and activity of the encoded protein. PMID- 10694980 TI - Molecular epidemiology of tuberculosis in the Nord Department of France during 1995. AB - In order to determine the current situation and to evaluate the human to human transmission of Mycobacterium tuberculosis in Northern France, the genetic polymorphism of strains was studied by using IS6110 fingerprint. One hundred and fifty-eight cases of bacteriologically confirmed tuberculosis were analyzed. One hundred and twenty-six patients (82%) were infected with genetically different isolates and 28 isolates (18%) were grouped into 14 clusters. No risk factors for recent Mycobacterium tuberculosis infections such as age, HIV status, immigrants, living in big cities were identified. This study shows that there was no major epidemic situation of tuberculosis in Northern France in 1995. Tuberculosis was characterized by a low proportion of HIV positive patients and a high proportion of elderly patients. PMID- 10694981 TI - Growth of a highly virulent strain of Mycobacterium tuberculosis in mice of differing susceptibility to tuberculous challenge. AB - Mycobacterium tuberculosis strain CDC 1551 exhibited unusually high levels of infectivity and virulence during a recent outbreak of tuberculosis in a rural community. Mice infected intravenously or aerogenically with M. tuberculosis CDC 1551 developed infections in the lungs and spleen which were almost identical with those for M. tuberculosis strains Erdman, H37Rv and Indian. There was also no significant difference in the survival rates of mice infected intravenously with CDC 1551, Erdman or H37Rv over a 3-month period. Studies designed to detect differences in the growth rates of CDC 1551 and Erdman in 3 inbred strains of mice failed to explain the high level of infectiousness and virulence expressed by CDC 1551 in human populations exposed to this organism. PMID- 10694982 TI - Human in vitro immune responses to Mycobacterium tuberculosis. AB - SETTING: T helper cells can be divided into 2 subsets on the basis of their cytokine generation. T helper 1 cells secreting gamma interferon and interleukin 2 appear to be more prominent in patients with limited tuberculous disease. OBJECTIVE: The purpose of this study was to evaluate human T helper cell immune responses to mycobacterial antigens in vitro and correlate these with the clinical features of patients with tuberculous infection or disease. DESIGN: We studied 51 subjects and 11 controls who were grouped according to disease involvement as follows: 1) Mantoux negative, BCG negative, no disease; 2) Mantoux positive, no disease; 3) localized extrapulmonary; 4) healed pulmonary; 5) active pulmonary; and 6) miliary/disseminated. Peripheral blood mononuclear cells were cultured with PHA, PPD or Tetanus Toxoid, proliferation assessed and the supernatant analysed using an ELISA for IFN gamma. ELISA was also used to measure M. tuberculosis specific antibodies in the serum. RESULTS: Mantoux size correlated with PPD proliferation r = 0.5, P = 0.005 and gamma IFN production r = 0.36, P < 0.01. All groups produced abundant gamma IFN although there was a trend toward higher production in groups 3 and 4. M. tuberculosis specific IgA (P = 0.003) and IgG1 (P = 0.002) was higher in groups 5 and 6. Those patients with limited disease (groups 2-4) had significantly lower levels of IgG4 than patients with severe disease (groups 5 & 6) (P < 0.02). CONCLUSION: In conclusion patients with healed or extrapulmonary disease have immune responses in vitro suggestive of a TH1 (cell mediated immune) response, whereas patients with miliary/disseminated disease have antibody production suggestive of a TH2 response, together with high gamma IFN production. Both TH1 and TH2 responses may be necessary for host protection if there is a high bacillary load. PMID- 10694983 TI - Virulence of recent notorious Mycobacterium tuberculosis isolates. PMID- 10694985 TI - Medical complications of diabetes mellitus in pregnancy. AB - Many women with diabetes develop complications of their chronic disease that may have a tremendous impact on their quality of life and their ultimate prognosis. Because Type 1 diabetes often begins at a very early age, it is quite common for women in their child-bearing years to be affected by these complications. As described in this article, diabetic complications and pregnancy may significantly affect each other, but it is not always easy to predict the course of either and to counsel these patients accordingly. Nevertheless, it appears that only in rare occasions should women with diabetes be advised against pregnancy, and that in most situations, with careful and knowledgeable management, a favorable outcome of pregnancy can be expected both for the mother and her infant. PMID- 10694984 TI - Diabetes and pregnancy: four motifs of modern medical history. PMID- 10694986 TI - Why do diabetic women deliver malformed infants? PMID- 10694987 TI - Role of diet and insulin treatment of diabetes in pregnancy. PMID- 10694989 TI - Obstetric management of pregnancies complicated by diabetes mellitus. PMID- 10694988 TI - The role of exercise in pregnant women with diabetes mellitus. PMID- 10694990 TI - Postpartum care of the woman with diabetes. AB - The postpartum period in women with diabetes or GDM allows both the physician and mother to relax from the intensive medical and obstetric management that has permitted, in most cases, a successful and joyous outcome. The role of the physician, however, must switch to a proactive and preventive mode to formulate a reproductive health plan for women with diabetes and GDM. The plan should be individualized to address glycemic management and surveillance, nutritional management, contraception prescription, future pregnancy planning, and lifestyle changes. Essential to the development of a reproductive health plan is the active participation of the patient, who through education gains an understanding of the far-reaching effects her active participation will have on her subsequent health and possibly on that of her future children. PMID- 10694991 TI - Physiologic and molecular alterations in carbohydrate metabolism during pregnancy and gestational diabetes mellitus. AB - Recent progress suggests that postreceptor mechanisms that contribute to insulin resistance of pregnancy appear to be multifactorial, but are exerted at the beta subunit of the insulin receptor and at the level of IRS-1. Gestational diabetes mellitus represents the combination of acquired and intrinsic abnormalities of insulin action. The resistance to insulin-mediated glucose transport appears to be greater in skeletal muscle from GDM subjects than from pregnancy alone. There is also a modest but significant decrease in maximal insulin receptor tyrosine phosphorylation in muscle from obese GDM subjects. Results also suggest that increased insulin receptor serine/threonine phosphorylation and PC-1 could underlie the insulin resistance of pregnancy and pathogenesis of GDM. Whether additional defects are exerted further downstream from IRS-1 remains to be investigated. PMID- 10694992 TI - Making the diagnosis of gestational diabetes mellitus. PMID- 10694993 TI - Management of gestational diabetes. PMID- 10694994 TI - Placental glucose transport in diabetic pregnancy. PMID- 10694995 TI - The infant of the woman with gestational diabetes mellitus. PMID- 10694996 TI - Cost analysis of diagnosis and treatment of gestational diabetes mellitus. PMID- 10694997 TI - Complementary and alternative medicine: a primer. PMID- 10694998 TI - Alternative therapies for menopause. AB - If a woman does not want to use, or cannot use, hormone replacement therapy, then she must consider other ways to address two issues related to menopause: reducing her risk of developing cardiovascular disease, osteoporosis, and other health problems that increase as women age, and symptomatology. Risk reduction of an array of health problems can be achieved through diet, exercise, and stress management. The nutraceuticals of specific vitamins, minerals, phytoestrogens, and essential fatty acid supplementations are a vital component of the risk reduction health program. Risk reduction of osteoporosis can be enhanced specifically through the use of ipriflavone and a comprehensive "bone building" vitamin and mineral program. Control of homocysteine levels for prevention of CAD, osteoporosis, and other health problems can be accomplished through B vitamin supplementation. The same interventions for risk reduction also may prove to be effective in prevention and treatment of menopausal-related symptoms, particularly when the B vitamins, magnesium, isoflavones, and essential fatty acids are used. If lifestyle interventions and nutraceuticals do not adequately address symptomatology, however, a woman has several alternative therapies from which to choose. There are numerous excellent multiherbal and homeopathic therapies that can be purchased over the counter. A woman also can choose to be evaluated by an alternative therapy practitioner and have a program designed specifically for her health needs. Although there has been limited clinical research of herbal and homeopathic alternative therapies for the menopause, when taken according to directions and if no contraindications exist, they have the potential for being extremely effective and safe options. PMID- 10694999 TI - Evaluation and management of insomnia in menopause. AB - Insomnia is a problem with complex and multifactorial etiologies that requires both standardized and individualized treatment interventions. Specific targets of treatment may include hyperarousal, poor sleep habits, underlying mood disorders, sedative overuse, pain and general medical problems, circadian dysrhythmias, sleep apnea, and restless legs syndrome. Optimal treatment also will incorporate stress management, coping strategies, enhancement of relationships, and promoting lifestyle changes that facilitate sleep. PMID- 10695000 TI - Menopause and cognitive function: estrogens and alternative therapies. PMID- 10695001 TI - Helping women help themselves: developing a menopause discussion group. AB - Menopause discussion groups are needed to provide women with accurate and up-to date information, but--more importantly--to facilitate sharing experiences, to help women understand their choices, and to empower them to be informed and take care of themselves. Members of a group who are comfortable expressing their feelings and thoughts find a therapeutic benefit from group participation. Health care providers are urged to develop these groups in their communities to assist women as they approach menopause and beyond. PMID- 10695002 TI - Hormone replacement therapy: making a 45-year plan. PMID- 10695003 TI - Diagnostic evaluation in gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) describes the clinical manifestations of reflux of gastric contents and the associated symptoms and patterns of tissue injury. Although its exact prevalence is difficult to determine, there is no doubt the GERD is the most common esophageal disease and probably among the most prevalent conditions seen in the primary care setting. GERD has a wide clinical spectrum, making the diagnostic evaluation challenging and complicated at times. Confirmatory test are rarely needed in patients with typical symptoms of heartburn or regurgitation who have a good clinical response to GERD therapy. This article describes the diagnostic tests necessary for some cases of GERD. PMID- 10695004 TI - Overview of medical therapy for gastroesophageal reflux disease. AB - There appears to be a hierarchy in the efficacy of therapies that are directed against GERD. A summary of this hierarchy, including therapies [table: see text] not approved by the U.S. Food and Drug Administration, is presented in Table 4. The individual practitioner must evaluate the appropriate point at which to place each patient on this hierarchy. Whether it is best to begin with the drug with the highest efficacy and step-down as possible for maintenance, never to step down, or to start with a less efficacious therapy and step up must also be individualized because there are no clear data to support a universal approach to all or even most GERD patients. PMID- 10695005 TI - Medical therapy. Management of the refractory patient. AB - Although relatively rare, GERD patients refractory to medical therapy remain a challenge for the clinician. Refractoriness can be diagnosed only if the patient is properly studied on medication and if what should be adequate medical therapy has been given a sufficient therapeutic trial. Use of 24-hour intragastric and intraesophageal pH-metry has improved the ability to manage patients who appear to be nonresponders. Simple advice and minor adjustments to medical therapy are usually all that is needed for most patients, but in some, management requires knowledge of the principles outlined in this article. If followed, only a small group of patients should be medically refractory. PMID- 10695006 TI - Extraesophageal gastroesophageal reflux disease. Presentations and approach to treatment. AB - Prevalence of gastroesophageal reflux disease (GERD) is common in the adult US population, but likely is underestimated as many patients present with symptoms other than heartburn or regurgitation. Ears, nose, throat, pulmonary, and cardiac symptoms also frequently are related to GERD. The diagnosis of GERD as a cause of these symptoms can be difficult and treatment strategies are much less clear than in patients presenting with heartburn or regurgitation. This article discusses the epidemiology, pathogenesis, diagnosis, and treatment of some of the manifestations of extraesophageal reflux disease. PMID- 10695007 TI - Peptic strictures of the esophagus. AB - Peptic esophageal strictures occur in the context of inadequately treated gastroesophageal reflux, especially in elderly patients. Studies show more pronounced abnormalities of esophageal function resulting in an increased number of prolonged reflux episodes. The diagnosis is best made by a combination of barium esophagram and endoscopy. Patients usually require esophageal dilation to relieve dysphagia followed by adequate medical therapy. Proton pump inhibitors are effective for preventing the recurrence of strictures after dilation. In young patients and patients with strictures that are difficult to dilate or need frequent dilations, surgery may be required; however, results can be disappointing. PMID- 10695008 TI - Endoscopy-negative gastroesophageal reflux disease. The hypersensitive esophagus. AB - Gastroesophageal reflux disease (GERD) is one of the mos common disorders of the gastrointestinal tract. Patients with GERD symptoms may exhibit a spectrum of endoscopic findings ranging from normal endoscopy (EGD negative) to severe ulcerative esophagitis. Recent evidence indicates that a large proportion of patients with GERD have normal endoscopy. The use of 24-hour ambulatory pH testing in the evaluation of symptomatic patients with EGD negative GERD allows further classification of these subjects into groups. Patients who have abnormal acid exposure and a positive symptom index constitute one group and patients who have normal acid contact time but record a convincing relationship between their symptoms and acid reflux on the pH analysis--positive symptom index--form another group. The latter group has been suggested to have a hypersensitive esophagus or "functional heartburn." This article reviews the prevalence, clinical features, origin of patient's symptoms, natural history, and treatment of patients with EGD negative GERD. PMID- 10695009 TI - Motility abnormalities in gastroesophageal reflux disease. AB - Esophageal dysmotility occurs in association with GERD; however, the cause of these motility abnormalities is not known. It is also not clear whether injury results from the presence of acid itself, inflammatory change or fibrosisin the esophageal wall. It is also unclear if reversal of these abnormalities takes place, and if so, to what degree. There are, however, a subset of patients who seem to have improvement with effective medical or surgical therapy, parodoxically, the same patients in whom a fundoplication, particularly a complete wrap, would lead to severe postoperative dysphagia secondary to preoperative dysmotility. What does all this mean for the individual patient? It is likely that most will not have any important change in esophageal motility abnormalities with standard medical or surgical therapy. Fundoplication might be safely performed in patients with minimal motility abnormalities, but those with severe abnormalities should be approached with caution. The conservative approach is to perform a partial fundoplication (Toupet) in those with ineffective motility (> 30% low-amplitude or nontransmitted contractions). It is hoped that future investigations will aid in understanding the pathogenesis of these abnormalities and how they can be used more precisely to guide antireflux therapy. PMID- 10695010 TI - Barrett's esophagus. Reducing the risk of progression to adenocarcinoma. AB - Barrett's metaplasia develops in 6% to 14% of individuals with gastroesophageal reflux. Barrett's adenocarcinomas are increasing in epidemic proportions for, as yet unknown, reasons; approximately 0.5% to 1% of patients with Barrett's metaplasia develop adenocarcinoma. Heartburn duration and frequency (but not severity), male gender, and white race are major risk factors for developing cancer. Obesity and smoking are weak risk factors. Survival is determined by depth of tumor invasion (stage). Once invasion of the muscularis propria occurs, most patients have developed widespread metastasis, even when clinical staging studies are negative. No currently available therapy results in prolonged survival once metastases develop. Thus, the more widespread use of effective surveillance strategies is the only currently available means for reducing the morbidity and mortality associated with Barrett's adenocarcinoma. PMID- 10695011 TI - Gastroesophageal reflux disease in children. AB - In the pediatric population, gastroesophageal reflux most often presents in infancy as effortless regurgitation, but pathologic GERD is accompanied by signs of malnutrition, respiratory diseases, and esophagitis or its complications. Because of the distinctive pathophysiology predisposing infants to GERD, the diagnostic approach must begin with a thorough history that determines the extent of further diagnostic tests and the course of management. Empiric therapy assumes importance in infants with GERD because of the limited differential diagnoses in consideration. Conservative therapy is of utmost importance because of the unique provocative factors in the pathophysiology of infantile GERD. Prokinetic pharmacotherapy takes precedence over acid suppression because of the more important role of motility factors compared with acid secretion in infantile GERD. PMID- 10695012 TI - Helicobacter pylori and gastroesophageal reflux disease. AB - The nature of the relationship between Helicobacter pylori (Hp) infection and gastroesophageal reflux disease (GERD) remains unclear. This article reviews the current body of knowledge regarding the association between these two common entities. The authors examine the potential interactions of Hp and GERD from epidemiologic and pathophysiologic viewpoints and summarize and critique the prevalence and eradication studies that have been performed to date. Special consideration is given to the possible effects that long-term use of proton pump inhibitors may have on Hp gastritis. PMID- 10695013 TI - Antireflux surgery. Indications, preoperative evaluation, and outcome. AB - Gastroesophageal reflux disease (GERD) is an extremely common disorder. Many patients require lifelong medical therapy for symptom control. In patients being considered for antireflux surgery, thorough evaluation is required. Laparoscopic antireflux surgery is a safe and effective method of treating patients who have severe, refractory, or complicated GERD. Excellent long-term results are obtained with minimal morbidity, freeing the patient from the burden of chronic medical therapy. PMID- 10695014 TI - Postfundoplication complications. Prevention and management. AB - Management of post-fundoplication problems begins by preventing complications from occurring. The prevention of complications after antireflux surgery can be divided into three important areas: (1) patient selection, (2) selection of the surgery, and (3) selection of the surgeon. After addressing prevention techniques, the author discusses the management of new postoperative symptoms such as Dysphagia, gas-bloat syndrome, and nausea and vomiting. PMID- 10695015 TI - [Mechanisms for resistance to anticancer agents and the reversal of the resistance]. AB - MDR results from overexpression of P-glycoprotein (Pgp) and multidrug resistance protein (MRP or MRP1) that function as ATP-dependent efflux pumps. Lung resistance related protein (LRP) is also supposed to be involved in MDR. The human canalicular multispecific organic anion transporter (cMOAT) gene that is responsible for the defects in Dubin-Johnson syndrome was isolated. cMOAT is homologous to MRP1 and supposed to be involved in drug resistance. Human cMOAT cDNA transfected LLC-PK1 cells, LLC/cMOAT-1, have increased resistance to vincristine (VCR), 7-ethyl-10-hydroxycamptothecin (SN-38), and cisplatin. The multidrug resistance (MDR)-reversing agents, cyclosporin A (CsA) and PAK-104P, almost completely reversed the resistance to VCR, SN-38 and cisplatin of LLC/cMOAT-1 cells by interacting with the substrate binding site of cMOAT. Treatment of human colorectal carcinoma SW-620 cells with sodium butyrate(NaB) induced LRP in the cells and conferred resistance to Adrianycin(ADM), VCR, VP-16, gramicidin D and taxol. Two LRP-specific ribozymes inhibited the NaB-induced expression of LRP in SW-620 cells and almost completely abolished their acquisition of the MDR phenotype. The accumulation of ADM, VCR and taxol was not decreased in NaB-treated cells, suggesting that ATP-binding cassette transporters are not involved in the MDR of NaB-treated cells. ADM was mainly located in the nuclei of untreated and the cytoplasm of NaB-treated cells. The accumulation level of ADM in the nuclei isolated from untreated cells or those from treated cells in the presence of anti-LRP polyclonal antibody was higher than that from treated cells in the absence of the antibody. Efflux of ADM from nuclei isolated from NaB-treated cells was enhanced compared with those from untreated cells and NaB-treated cells transfected with a LRP-specific ribozyme. The polyclonal antibody against LRP inhibited the enhanced efflux of ADM from nuclei isolated from NaB-treated cells. These findings indicate that LRP is involved in resistance to ADM, VCR, VP-16, taxol and gramicidin D, and has an important role in the transport of ADM from the nucleus to the cytoplasm. PMID- 10695016 TI - [Gene therapy for inherited diseases using heamatopoietic stem cells--gene therapy for patients with chronic granulomatous disease]. AB - The possibility of gene therapy for inherited diseases with a single gene mutation in Figure 1 had been verified by the successful treatment with bone marrow transplantation. As the gene therapy method and theory has been progressing rapidly, it is expected that gene therapy will overcome the complications of bone marrow transplantation. Of these inherited diseases, chronic granulomatous disease (CGD) is the one of the most expected disease for gene therapy. CGD is an inherited immune deficiency caused by mutations in any of the following four phox genes encoding subunits of the superoxide generating phagocyte NADPH oxidase. It consists of membranous cytochrom b558 composed of gp91 phox and p22 phox, and four cytosolic components, p47 phox, p67 phox, rac p21 and p40 phox, which translocate to the membrane upon activation. In our group study, more than 220 CGD patients has been enrolled. The incidence of CGD patients was estimated as 1 out of 250,000 births. The expected life span of the CGD patients is 25 to 30 years old by the Kaplan Meier analysis. Comparing with the ratio of CGD subtype in US and Europe, that with p47phox deficiency is lower (less than 10%/o vs. 23%) and that of gp91 phox deficiency is higher (more than 75% vs. 60%). Prophylactic administration of ST antibiotics and IFN-gamma and bone marrow transplantation have been successfully employed in our therapeutic strategy. However, it is necessary to develop the gene therapy technology for CGD patients as more promising treatment. In the current study we constructed two retrovirus vectors; MFGS-gp91/293 SPA which contains only the therapeutic gp91 phox gene, a bicistronic retrovirus pHa-MDR-IRES-gp91/PA317 which carries a multi drug resistant gene (MDR1) and the gp91phox gene connected with an internal ribosome entry site (IRES). We demonstrate high efficiency transduction of gp 91 phox to CGD EB virus established cell line with high levels of functional correction of the oxidase by MFGS-gp91 and by pHa-MDR-IRES-gp91, respectively. We also demonstrate sufficient transduction of gp91 phox to CD34+ hematopoietic stem cell from the patients with gp91 phox deficiency by MFGS-gp91/293 SPA. Our current studies suggest that the combination of the 293-SPA packaging system and the bicistronic retrovirus system inserted MDR1 gene make our CGD gene therapy more feasible for clinical application. PMID- 10695017 TI - [Gene therapy of cardiovascular disorders]. AB - More than 300 protocols have been developed for human gene therapy, but, it has not yet been proved to be a successful therapeutic strategy. One of the most important barriers to success is the development of efficient gene delivery systems. We have developed HVJ-liposomes by combining fusion proteins of HVJ (Hemagglutinating virus of Japan; Sendai virus) with liposomes containing DNA. This vector system has been very effective for in vivo gene delivery, especially in cardiovascular systems. Using HVJ-liposomes, we have reported successful gene therapy experiments such as prevention of restenosis after balloon injury, suppression of dysfunction of vein graft, and experimental ischemic disorders. Indeed, the success in the treatment of arteriosclerosis obliterance by VEGF (vascular endothelial growth factor) gene transfer was reported recently. These cardiovascular gene therapy strategies appear to be very promising therapeutics in future. PMID- 10695018 TI - [Gene therapy using anticancer drug-resistance genes]. AB - Myelosuppression is a major dose-limiting factor in cancer chemotherapy. Introduction of drug-resistance genes into bone marrow cells of cancer patients has been proposed to overcome this limitation. In theory, any gene whose expression protects cells against the toxic effects of chemotherapy should be useful in vivo for this purpose. Among such genes, human multidrug-resistance gene (MDR1) has been studied most extensively for this purpose, and clinical trials of drug-resistance gene therapy have been started in the US for cancer patients who undergo high-dose chemotherapy with autologous hematopoietic stem cell transplantation. In Japan, our clinical protocol of MDR1 gene therapy "A clinical study of drug-resistance gene therapy to improve the efficacy and safety of chemotherapy against breast cancer" has been submitted to the government. To improve the efficacy and safety of this drug-resistance gene therapy, we have constructed a series of MDR1-bicistronic retrovirus vectors using a retrovirus backbone of Harvey murine sarcoma virus and internal ribosome entry site (IRES) from picornavirus to co-express a second gene with the MDR1 gene. MDR1-MGMT bicistronic vectors can be used to protect bone marrow cells of cancer patients from combination chemotherapy with MDR1-related anticancer agents and nitrosoureas. In addition, MDR1-bicistronic retrovirus vectors can be designed to use the MDR1 gene as an in vivo selectable marker to enrich the transduced cells which express therapeutic genes, if disease is curable by the expression of a single-peptide gene in any types of bone marrow cells or peripheral blood cells. PMID- 10695019 TI - [Gene therapy for acute hepatitis using CrmA gene transduction]. AB - Fas-Fas ligand interactions between adenovirus-infected hepatocytes and cytotoxic T lymphocytes (CTLs) play a major role in killing of vector-transduced hepatocytes. Cytokine response modifier A (CrmA) is known to protect lymphoid cells from Fas-mediated apoptosis by inhibiting caspase 8. In this study, we generated an E1-deleted adenovirus expressing CrmA, and investigated the effect of exogenous CrmA expression on the inhibition of Fas-mediated apoptosis in murine hepatocytes in vitro and on the prolongation of the transgene expression in adenovirus-transduced hepatocytes in vivo. Agonistic anti-Fas antibody induced massive apoptosis into hepatocytes, however, most of the cells expressing CrmA were escaped from apoptosis and survived. This result showed that anti-apoptic function was obtained in murine hepatocytes by expressing CrmA. The prolongation of the transgene expression was studied using mice with congenital deficiency of lysosomal beta-glucuronidase (GUSB). The serum GUSB activity from the mice injected only an adenovirus expressing human beta-glucuronidase disappeared within 70 days, however, significant GUSB activity was observed for more than 130 days in the mice co-transduced with adenoviruses expressing both GUSB and CrmA. Moreover, histochemical analysis showed GUSB expressions in the liver even at 130 days after the viral administration. These observations demonstrate that the prolongation of the transgene expression can be achieved in rodent liver by CrmA co-expression using adenoviral gene transfer. PMID- 10695020 TI - Establishment and characterization of cell lines derived from serous adenocarcinoma (JHOS-2) and clear cell adenocarcinoma (JHOC-5, JHOC-6) of human ovary. AB - The cell lines designated JHOS-2, JHOC-5 and JHOC-6 were established from epithelial ovarian carcinomas. JHOS-2 was established from a serous adenocarcinoma of a 45-year-old Japanese woman, JHOC-5 from a recurrent tumor of a clear cell adenocarcinoma of a 47-year-old Japanese woman and JHOC-6 from a tumor of a clear cell adenocarcinoma of a 43-year-old Japanese woman. These cell lines have grown well and serial passages were successively carried out more than 20 times. The monolayer cultured cells revealed neoplastic and pleomorphic features, and grew in multilayers. Electron micrographs revealed epithelial origins that had desmosomes and tonofilaments. PMID- 10695021 TI - Establishment and characterization of human ovarian clear cell adenocarcinoma cell line (SMOV-2), and its cytotoxity by anticancer agents. AB - A novel cell line derived from a surgically resected ovarian clear cell adenocarcinoma of 46 year-old Japanese woman was established and designated SMOV 2. Cells of this lineage were continuously propagated in vitro over 44 months and were grown in a mono-layered sheet with a doubling time of 48.2 hours. The histopathology of the transplanted tumor in nude mice showed two distinctive cell types, hobnail cells and clear cells, which demonstrated recognizable characteristics of clear cell adenocarcinoma, as compared to resected original tumors. At the molecular level, SMOV-2 cells had the wild type p53 genes that were free from missence mutations. Anticancer agents (cisplatin and paclitaxel) were examined for cytotoxity against these SMOV-2 cells in vitro. These examinations revealed that the chemotherapy-treated cells had decreased proliferation, cell cycle arrests, and induction of apoptosis by the anticancer agents. As can be gleaned from this research, SMOV-2 is a valuable model to study the mechanism of apoptotic responses of solid tumors to future anticancer agents. PMID- 10695022 TI - Efficient DNA fingerprinting method for the identification of cross-culture contamination of cell lines. AB - In order to identify cross-culture contamination of cell lines, we applied DNA fingerprinting using variable number of tandem repeat (VNTR) loci and short tandem repeat (STR) loci amplified by polymerase chain reaction (PCR) instead of a radioisotope labeled multilocus probe. Eleven cell lines were used for the Apo B and D1S80 loci detection, and twelve cell lines were examined in the Y chromosome analysis. The data obtained from the sister cell lines NALM-6 and B85, two MOLM-1 cultures from two cryopreserved tubes, and four subclones of BALM-9 and its sister cell line BALM-10, displayed clear and distinct bands of each PCR product for both Apo B and D1S80. Detection of a Y-chromosome DNA sequence is another very informative marker for the identification of cell lines, if the Y chromosome is present. We examined eight cell lines for the expression of four STR loci; the data thus generated were compared with the results previously reported from other laboratories. The resulting electrophoretic banding patterns showed that our "home-made" STR detection system is a useful and efficient tool for the authentication of cell lines. PCR detection of VNTR and STR loci represents a simple, rapid and powerful DNA fingerprinting technique to authenticate human cell lines and to detect cross-culture contamination. This PCR technique may be used in lieu of the more time-consuming, labor-intensive and radioactive Southern blot multilocus method. PMID- 10695023 TI - [Reliability of disc-diffusion susceptibility testing for arbekacin, vancomycin and teicoplanin against methicillin-resistant Staphylococcus aureus]. AB - We investigated the differences in judgments among four disc-diffusion methods on susceptibility testing of arbekacin (ABK), vancomycin (VCM) and teicoplanin (TEIC) against 37 strains of methicillin-resistant Staphylococcus aureus (MRSA). These results were compared with minimum inhibitory concentrations (MICs) obtained from micro broth-dilution method. A marked difference was noted in the judgment of susceptibility to TEIC in Tri-disc method, that is 2 strains (5.4%) fell into sensitive (+3) 34 strains (91.9%) into moderately sensitive (+2) and 1 strain (2.7%) into moderately resistant (+), while in Sensi-disc method all strains fell into sensitive (S). According to the MICs, no strain of the MRSA tested revealed resistance to ABK, VCM and TEIC. Consequently, these three antimicrobial agents were thought to be effective on MRSA infections. From these results, we concluded that Tri-disc method for glycopeptide against MRSA, especially for TEIC, is not recommendable as a disc-diffusion method in susceptibility testing. PMID- 10695024 TI - [In vitro activity of biapenem (BIPM) against clinically isolated respiratory pathogens in 1996-1998]. AB - The in vitro antibacterial activity of biapenem (BIPM), a new carbapenem antibiotic, was compared with those of imipenem (IPM), panipenem (PAPM), meropenem (MEPM), ceftazidime (CAZ) and piperacillin (PIPC) against 280 isolates of 9 respiratory pathogens. The MIC90s of biapenem (BIPM) for methicillin susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pneumoniae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Haemophilus influenzae were 0.12, 32, 0.25, 0.06, 4 and 8 micrograms/ml, respectively. In comparison with other antibiotics, the activity of biapenem (BIPM) for P. aeruginosa was as potent as meropenem (MEPM), but for H. influenzae it was slightly less than those of other antibiotics, and for other respiratory pathogens it was as potent as those of other antibiotics. The MIC90s of biapenem (BIPM) for Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae and Serratia marcescens were 0.06, 1, 1, 0.5 microgram/ml, respectively, and which were equal to or somewhat lower than those of other antibiotics. Biapenem (BIPM) showed strong activity against Gram-positive and Gram-negative pathogens, especially P. aeruginosa in general. Based on these results, biapenem (BIPM) is seemed to be highly useful antibiotic for the treatment of respiratory infections with several organism. PMID- 10695025 TI - [Antimicrobial activities of meropenem against clinically isolated strains in 1997]. AB - In order to evaluate antimicrobial activity of meropenem (MEPM), minimum inhibitory concentrations (MICs) of MEPM and control drugs were determined against clinical isolates in 1997. The results were as follows; 1. Antimicrobial activities of MEPM against Gram-positive bacteria were stronger than those of cephems (CEPs) and were approximately equal to those of imipenem (IPM) and panipenem (PAPM). 2. Carbapenems showed strong antimicrobial activities against Enterobacteriaceae, Glucose non-fermentative Gram-negative rods and Bacteroides fragilis group that were multiple drug resistant including the third generation CEPs. Antimicrobial activities of MEPM against these organisms were stronger than those of IPM and PAPM. By comparing antimicrobial activities of MEPM against Gram negative bacteria in 1997 with those obtained in 1993, increase of resistance was not observed. 3. MIC-ranges of MEPM were low against the resistant strains of Pseudomonas aeruginosa to IPM and PAPM. It was considered that these resistant strains were not expressing oprD products (D2 porin protein), forming main outer membrane porin channels of carbapenems and basic amino acids. PMID- 10695026 TI - [Pharmacotherapy of depression in the elderly]. AB - Antidepressants are not only effective in younger adults but also in older persons with depression. However there are peculiarities, which have to be taken into account when treating older patients. The enhanced risk of side effects has to be balanced with the more serious consequences of an untreated depression in old age. The efficacy in older patients has been proven not only for tricyclic antidepressants but also for SSRI and other newer antidepressants. The easy handling, the side effect profile and the safety in overdose are advantages of SSRI and newer antidepressants, which become particularly important in old age. The advantages and disadvantages of different classes of antidepressants with regard to comorbidity and comedication are discussed. In older subjects, changes of dosage should be slower and for some, but not for all antidepressants, dosage should be lower. PMID- 10695027 TI - [Detection of neurotransmitter interactions with PET and SPECT by pharmacological challenge paradigms]. AB - Functional brain imaging with positron emission tomography (PET) and single photon emission computerized tomography (SPECT) enables the in vivo study of specific neurochemical processes in the context of normal regulatory mechanisms and pathophysiological alterations of the brain. By combining these methods with pharmacological challenge-paradigms, the study of functional interactions of different neurotransmitter systems is possible. This review will present data from animal and healthy volunteer studies as well as first data from investigations in different patient populations with regard to this research direction. Especially, interactions of different neurotransmitter systems with the dopaminergic and the cholinergic system will be discussed. The database acquired so far confirms existing models of neuronal feedback-circuits, and the first clinical results are consistent with the hypothesis of an increased dopaminergic responsivity in schizophrenic patients. These results open up new perspectives for a further evaluation of treatment response predictors from drug challenge studies and for the development of new drug treatments for neuropsychiatric disorders. PMID- 10695028 TI - [Cognitive remediation. A new chance in rehabilitation of schizophrenic disorders?]. AB - A critical review of current approaches and principles of cognitive remediation strategies in rehabilitation of schizophrenia is given. Selection of cognitive functions targeted in compensatory training programs could be based on results of neuropsychological predictor research on social and vocational functioning in community and neuropsychological rate limiting factors in rehabilitation. Methodological flaws in data base, missing of task analysis of more complex skills like social perception, social skills and interpersonal problem solving and the lack of evaluation of training generalization on work performance are discussed. Finally the cognitive remediation program developed in the Department of General Psychiatry and Psychotherapy/University of Freiburg, Germany is proposed. The components focused on training in attention, memory, and executive function (decision making, planning). Compensatory strategy building, and computer-mediated automatization are integrated in a group setting. PMID- 10695029 TI - [Effect of activation tasks on acute neuroleptic-induced akathisia]. AB - In this study, we investigated experimentally the effects of different activation procedures on both motor and psychic symptoms in of 11 in-patients with acute neuroleptic-induced akathisia using the Hillside and Barnes akathisia rating scales and videotape rating technique. Motor activation was achieved by finger tapping. Cognitive activation tasks consisted of sequences of mental calculations which were designed either to be easy to perform or to produce stress due to a given time limit or to more difficult calculation operations, respectively. Motor as well as psychic symptoms of akathisia decreased during both motor and simple cognitive activation without stress. By contrast, stress-producing calculation tasks led to an increase in motor and psychic symptoms immediately following the task performance. These possibly specific effects of activation procedures on symptoms might be useful in differentiating acute neuroleptic-induced akathisia from other neuroleptic-induced and extrapyramidal movement disorders. PMID- 10695030 TI - [Clinical aspects of "argyrophilic grain disease"]. AB - Argyrophilic grain disease (AGD) is a frequently occurring degenerative illness of the aging human brain. It is accompanied by progressive pathological alterations of the cytsokeleton which are traceable to an abnormal phosphorylation of the microtubule associated tau protein. Histologically, it is possible with the help of suitable staining techniques to identify pathognomonic spindle-shaped cellular inclusions (argyrophilic grains). These cellular inclusions display a typical cortical as well as subcortical distribution pattern. The goal of the present study is the retrospective evaluation of the clinical findings from 53 individuals with neuropathologically demonstrable AGD related changes of the brain. Nearly one-half of the cases (49%) was classifiable as demented in accordance with DSM IV-criteria. Moreover, the frequency of the dementia increased significantly in relation to the growing severity of the AGD associated pathological cytoskeletal degeneration. These results confirm the assumption that AGD can cause degenerative changes ranging from cognitive impairment all the way to dementia. They also underscore the necessity of further prospective studies pertaining to the clinical aspects of this still enigmatic disease. PMID- 10695031 TI - [Early manifestation of fronto-temporal dementia]. AB - The term frontotemporal dementia is used to describe a primary degenerative form of dementia, which is characterized by typical clinical, neuropsychological, radiological and neuropathological features. Its onset is usually before the age of 65 years; manifestations before the age of 30 years have rarely been described. We report the case of a 22-year old man, who showed symptoms of behavioural disorder such as social retreat, lack of initiative, mental rigidity, progressive reduction of speech, and stereotyped behaviour. The neuropsychological examination revealed disorders of the executive functions. The cerebral MRI investigations showed bifrontal atrophies corresponding with hypoperfusion areas on the SPECT. Other investigations including EEG, evoked potentials, duplex ultrasonography, cerebral angiography, laboratory tests and cerebrospinal fluid were normal. In the present case report we discuss the clinical presentation of frontotemporal dementia with early onset. PMID- 10695032 TI - [Propane abuse. Extreme dose increase due to development of tolerance]. AB - In spite of its serious sequelae, volatile substance abuse (VSA) attracts very little public attention in Germany. Our case report describes an adult male who inhaled propane for recreational purposes. Initially, he achieved short-lived euphoria and hallucinations. He compensated for the developing tolerance by increasing the dosage, finally consuming 5 litres of fluid propane daily. Getting such quantities was facilitated by his occupational access to propane. Since he abused the propane in an apartment house, he also exposed third parties to the danger of explosion. Clinical examination revealed disturbances in orientation, restricted perceptivity and concentration, reduced mnemonic performance, and psychomotor agitation. All these symptoms diminished during a 6-month follow-up. The relationship of his organic mental disorder to the abuse of propane was not clear, since he had also abused alcohol. PMID- 10695033 TI - [The dopamine hypothesis of schizophrenia. New findings for an old theory]. AB - The dopamine hypothesis of schizophrenia was based on the neuroleptical blockade of central dopamine D2 receptors. Brain imaging studies, however, generally failed to demonstrate a significant increase in central D2 receptors among schizophrenic patients. Using a novel approach, the group of Laruelle and Abi Dargham was now able to demonstrate that schizophrenic patients have increased synaptic dopamine concentrations in the striatum. Endogenous dopamine competes with a radioligand for binding at dopamine D2 receptors; compared to healthy control subjects, blockade of dopamine production in neuroleptic-naive schizophrenic patients induced a significantly higher increase in D2 binding of the infused radioligand, indicating higher endogenous synaptic dopamine. A similar increase in D2 binding was also observed in drug-free schizophrenics who had previously been treated with neuroleptics; these patients also showed an increased density of striatal D2 receptors, most likely due to counteradaptive upregulation of D2 receptors during neuroleptic medication. The Columbia study provides an important indication of hyperdopaminergic function in schizophrenia. PMID- 10695034 TI - [Omega-3 fatty acids in psychiatry]. AB - Omega-3 fatty acids (ALA, EPA, DHA) are essential polyunsaturated fatty acids. Due to their pivotal involvement in signal transduction processes in the CNS, a role for these fatty acids in psychiatric disorders has been postulated. This review summarizes the latest findings on the physiological function of these compounds in the CNS and gives a comprehensive overview on the emerging therapeutic role of these psychoactive drugs in psychiatric disorders, with special emphasis being put on affective disorders and schizophrenia. PMID- 10695035 TI - [Cytokines in viral infections]. AB - The role of cytokines and chemokines in viral replication and antiviral immunity is discussed. Some viruses may have genes that mimic the host genes (cytokines or cytokine receptors) and their functioning may inhibit the synthesis or processing of cell proteins important for defense. Some of host cytokines may be involved in the pathogenesis of viral infection (HIV, herpes virus, cytomegalovirus, and some others). PMID- 10695037 TI - [Serological study of hepatitis C virus NS5 protein epitopes and their clinical and diagnostic significance]. AB - Three peptides corresponding to the 2295-2317 aa NS5 HCV region and individual parts of this region were synthesized. Antigenic properties of these peptides were investigated. The 2295-2317 aa region contains at least two epitopes of different nature. The full-sized peptide is more promising for the diagnostic studies. Optimal conditions for ELISA with this peptide were defined, allowing the maximum complete utilization of the potentialities of both epitopes. PMID- 10695036 TI - [Distribution and features of infection with hepatitis viruses B and C during hemodialysis treatment]. AB - The prevalence of hepatitides B and C was evaluated in 140 patients treated by hemodialysis. Almost half of patients (48%) had acute hepatitis B which completely resolved. Acute hepatitis B was detected in 6% in the course of observation. In 6% chronic hepatitis B was diagnosed, and in 24% chronic hepatitis C. A combination of hepatitides B and C was diagnosed in 2% patients. Only 12% patients were not infected with hepatitis. Genotype 1b predominated in patients with HCV infection (73%); genotypes 1a, 21, and 3a were equally incident (9%). Replication of HBV and HCV in patients with uremia under conditions of hemodialysis was detected in 83 and 86% patients, respectively. Relationship between HBV and HCV infection and the duration of hemodialysis treatment was analyzed. The percentage of non-infected patients persistently decreased, and the time course of HBV and HCV infection was different. Infection with HBV after the beginning of hemodialysis occurred sooner (16.0 +/- 4.0 months) than with HCV (30.2 +/- 4.6 months, p < 0.04). The levels of SGPT and SGOT in patients with various manifestations of HBV and HCV infection treated by hemodialysis were followed up. PMID- 10695038 TI - [Clinico-laboratory characteristics of viral hepatitis C with various HCV genotypes]. AB - Clinical and biochemical features of the acute phase of icteric hepatitis C in various HCV genotypes have been studied. The HCV genotypes determine the duration of incubation period and some clinical signs in the preicteric and icteric periods of acute hepatitis C. The biochemical picture and formation of chronic hepatitis virtually did not depend on the virus genotype. PMID- 10695039 TI - [Gene analysis and phenotypic characteristic of highly-reproductive reassortants, containing the gene for bird influenza virus subtype H2 hemagglutinin]. AB - A series of reassortant clones with antigenic formulae H2N1 and H2N3 were produced by genetic reassortment performed with the use of an avian influenza virus, A/Pintail Duck/Primorie/695/76 (H2N3) and a high-yield reassortant strain X-67. Preliminary identification of the parent origin of NP and NS genes for 5 reassortants was performed by comparison of the mobilities of virus-specific proteins in polyacrylamide gel electrophoresis. The parent origin of genes of internal and nonstructural proteins for 3 reassortants was identified by partial sequencing. Although the genes of internal and nonstructural proteins of the reassortants originated from high-yield X-67 virus, only H2N3 reassortants were similar to the high-yield parent virus as concerns the level of the virus accumulation evaluated by hemagglutination titration and measurement of the virus protein content. PMID- 10695040 TI - [Production of interleukin-2 in vitro and in vivo in elderly people, vaccinated with live and inactivated flu vaccines separately and combined]. AB - Forty-three elderly individuals were immunized with Russian trivalent live cold adapted influenza vaccine (LIV) and US trivalent influenza vaccine (IIV) administered separately or in combination. IL-2 production in vitro (in supernatants of cultures of lymphocytes stimulated with homologous viral antigens and PHA) and in vivo (in blood serum) and other factors of specific antiinfluenza immunity were compared. Vaccination of elderly subjects with commercial vaccines induced T-helper immunological memory, which manifests by increased secretion of IL-2 in vitro and in vivo. Simultaneous vaccination with LIV + IIV and revaccination (in 1 month) with LIV was the most effective method stimulating IL 2 production. The levels of IL-2 production in vitro were in good correlation with the secretion of this cytokin in vivo, lymph proliferation, and serum antibody production. No correlation between IL-2 production in vitro and the formation of local immune response (IgA in nasal swabs) was detected. PMID- 10695041 TI - [Mechanisms for the therapeutic effect of herpes polyvaccines in chronic ophthalmic herpes and genital herpes]. AB - Commercial inactivated culture polyvaccine against herpes simplex viruses (types 1 and 2) developed at D. I. Ivanovsky Institute of Virology promoted cessation of viremia. During the first vaccination viremia coincided with appearance of a focal allergic test on the retina, which is proposed for the diagnosis of herpetic involvement of the posterior compartment of the eye. T-cellular immunity normalized after a course of vaccination. Experimental immunization of rats and vaccination of patients with chronic ophthalmic and genital herpes demonstrated the therapeutic activity of inactivated herpetic polyvaccine in suppositoria. PMID- 10695042 TI - [Protective effect of a new antiviral preparation of phosprenyl in experimental tick-borne encephalitis]. AB - Antiviral activity of phosprenyl was studied in BALB/c mice infected with tick borne encephalitis (TBE) virus. Up to 60% animals infected with TBE virus survived after 1-3 intramuscular injections of phosprenyl. The mortality in the untreated group infected with the virus was 100%. Direct antiviral effect of phosprenyl was studied in sensitive SPEV cells infected with TBE virus. The titer of the virus decreased 10-fold in the cells treated with the drug vs. untreated control cells. Phosprenyl stimulates some interleukins: gamma-interferon, tumor necrosis factor-alpha, and interleukin-6. The stimulating effect of the drug manifests in intact animals and in those infected with TBE virus and treated with phosprenyl. The prospects of further trials of the drug as a therapeutic and prophylactic agent in TBE are discussed. PMID- 10695043 TI - [Epidemic outbreak of meningitis and meningoencephalitis, caused by West Nile virus, in the Krasnodar territory and Volgograd region (preliminary report)]. AB - Sera from 102 inpatients from the Volgograd region (64) and Krasnodar region (38) were tested for antibodies to West Nile (WN) virus in hemagglutination inhibition (HI) test and for IgM and IgG antibodies in enzyme immunoassay (EIA). Diseases etiologically associated with WN virus were diagnosed in 81 patients: in 50 out of 64 in the Volgograd region and in 31 out of 38 in the Krasnodar region, which makes 79.4%. Specificity of antibodies to WN virus was confirmed in HI and EIA with WN antigens, related flaviviruses (Japanese encephalitis and yellow fever), and Sindbis alfavirus. A considerable number and the incidence of WN infection suggest that an epidemic caused by WN virus occurred in the Krasnodar and Volgograd regions in summer 1999. PMID- 10695044 TI - [Characteristics of "Avaxim" hepatitis A vaccine produced by the firm "Pasteur Merrier" (results of field clinical trials)]. AB - Controlled field clinical trials of Avaxim vaccine from hepatitis A (Pasteur Merrier-Connot) were carried out in adults and children aged 3-14 years to evaluate its reactogenicity and antigenic properties. The vaccine was weakly reactogenic both in adults and children. A single injection of the vaccine resulted after 1 month in the production of anti-HAV antibodies in 77.4% initially seronegative adults and 94.5% children. In adults the mean geometrical titer of antibodies was 95 mIU/ml and in children 165 mIU/ml, which was 5-8 times higher than the protective titer. These data recommend Avaxim vaccine for practical public health. PMID- 10695045 TI - [Induction of baculovirus infection at various stages of ontogenesis of the silkworm (Ocneria dispar L.)]. AB - The inducibility of baculovirus infection in Ocneria dispar L. of West Siberian population is studied. The insects are rather stable to induced polyhedrosis. PMID- 10695046 TI - [On vaccine therapy of chronic urogenital chlamydia infection]. AB - The efficiency of vaccine therapy of a small group of patients with chronic urogenital chlamydial infection can be regarded as preliminary results. The group included only patients treated with antibiotics many times without success; vaccine therapy led to stable cure and elimination of Chlamydia from the organism. Therefore, vaccine therapy of the above mentioned patient population is justified. PMID- 10695047 TI - The true history of the discovery of penicillin, with refutation of the misinformation in the literature. AB - In 1928, while investigating variant strains of Staphylococcus aureus, Alexander Fleming found that a fungus growing near the edge of a culture of S. aureus produced a substance which had diffused into the medium, lysed the nearest organisms and thus produced a clear area immediately surrounding the fungus. Further away was a zone of transparent, degenerate colonies, while those in the outer zone appeared healthy. This culture plate was one of several left on his laboratory bench at room temperature while he was away on holiday. On his return, he noticed the difference on this plate from the usual contaminated plate. He cultivated the fungus, Penicillium notatum, which he was initially informed was P. rubrum, and called the soluble extract penicillin. It cured local infections but, at the time, could not be purified to treat systemic infections. Many authors have criticized apparent failings in Fleming. This paper catalogs the criticisms and provides evidence to refute them. PMID- 10695048 TI - Chlamydia trachomatis infection in pregnancy: risk factor for an adverse outcome. AB - A cohort of 122 pregnant women attending the hospital antenatal clinic in northern India were studied to determine the prevalence of genital chlamydial infection, and any adverse effect on the pregnancy. Endocervical swabs were taken at > 12 weeks of pregnancy and cultured for Chlamydia trachomatis. Twenty-six (21.3%) pregnant women were found to be infected with C. trachomatis. The mean age, gravidity and parity were significantly higher (25.03 vs 23.6 years, 1.88 vs 1.72 and 0.92 vs 0.68 respectively [P < 0.005]) in women from whom C. trachomatis was isolated. Follow-up was possible in 87 women who delivered in the hospital. There was increased incidence of still-birth, prematurity and low birth-weight in the C. trachomatis-positive women (16.6% vs 5.7%, 26.6% vs 18.4% and 26.6% vs 23.0%), and these differences were statistically significant (P < 0.5, P < 0.5 and P < 0.05 respectively). The results suggest a definite need for C. trachomatis screening on a wider scale, both in different risk groups of asymptomatic antenatal women and in neonates, to confirm these findings. PMID- 10695049 TI - Characterisation of yeasts implicated in vulvovaginal candidosis in Irish women. AB - High-vaginal swabs were taken from 98 women attending a genitourinary clinic in Dublin city who presented with symptoms of vaginitis. Twenty-eight (28.6%) proved culture-positive for yeasts. Candida albicans was isolated from 26 of the yeast positive patients and Candida glabrata and Candida tropicalis were isolated from a further two patients. Two patients were colonized by two yeasts simultaneously (C. albicans and Candida lucitaniae, and C. albicans and Candida krusei). Yeasts were characterised on the basis of their adherence ability, extracellular enzyme production and resistance to antifungal agents. All C. albicans isolates demonstrated high adherence abilities to buccal epithelial cells, but produced relatively low levels of phospholipase and acid proteinase. The non-C. albicans isolates demonstrated low levels of adherence, but no extracellular enzyme production was detected. Isolates were most sensitive to the polyenes amphotericin B and nystatin but a large proportion (50%) of C. albicans isolates were resistant to clotrimazole, which is an important agent in the treatment of vulvovaginal candidosis. PMID- 10695050 TI - Enteric pathogens and their isolation at a cancer hospital in Pakistan. AB - Diarrhoeal diseases remain a major cause of mortality and morbidity in developing countries. However, due to lack of funds, supply problems and some inexperience, some laboratories have difficulty identifying a causative agent in stool samples. In the year following the opening of the Shaukat Khanum Memorial Cancer Hospital and Research Centre in Lahore, Pakistan, the microbiology department had not isolated a single enteric pathogen. From January 1996, new culture techniques were introduced, with a resulting increase (10%) in identification of these pathogens. In addition, the introduction of formol-ether concentration made a significant contribution to the number of intestinal parasites seen. This report demonstrates how simple microbiology methods made a difference to the running of the department and, ultimately, to the patients. PMID- 10695051 TI - Dietary tomato and grape pomace in rats: effect on lipids in serum and liver, and on antioxidant status. AB - Addition of tomato and grape pomace to the cholesterol (0.3%) diet of male Wistar rats produced a dose-dependent effect. During the eight-week experiment, 5% pomace showed no effect; however, 15% pomace reduced serum cholesterol levels from 4.4 mmol/L to 2.5 mmol/L (tomato) and 2.0 mmol/L (grape). At a concentration of 15%, both tomato and grape pomace induced a redistribution of cholesterol in lipoproteins, resulting in a pronounced anti-atherogenic profile: reduced cholesterol concentration in very-low-density lipoprotein (VDL) (24% [tomato], 50% [grape]) and low-density lipoprotein (LDL) (3-fold, 3.6-fold). In addition, grape pomace increased cholesterol concentration in high-density lipoprotein (HDL) by 26%. Both types of pomace reduced the VLDL and LDL contribution to cholesterol transport in favour of HDL. Grape pomace (15%) produced a significant reduction in cholesterol and triacylglycerols in the liver and serum respectively. Diets containing tomato and grape pomace reduced plasma levels of conjugated dienes by 30-50%, and showed a tendency towards higher superoxide dismutase and glutathione peroxidase activity in the liver. PMID- 10695052 TI - Expression of Bcl-2 family proteins during chemotherapeutic agents-induced apoptosis in the hepatoblastoma HepG2 cell line. AB - This study demonstrates that two anticancer drugs, taxol and doxorubicin (Dox), can kill human hepatoblastoma HepG2 cells in a dose-dependent manner via the induction of apoptosis. Characteristic events, including externalization of phosphatidylserine, cytoplasmic shrinkage, chromatin condensation and DNA degradation, were observed in a large majority of the drug-treated cells. DNA fragmentation showed that a ladder of DNA fragments of approximately 200 bp multiples was observed in taxol-treated, but not in Dox-treated, cells. In addition, the expression patterns of Bcl-2 family members during taxol or Dox treatment were investigated. Results from Western blot analysis indicated that HepG2 cells did not express either the death repressor Bcl-2, or the death promoters Bcl-XS and Bax. However, during the apoptotic process one death repressor, Bcl-XL, and two death promoters, Bak and Bad, were expressed. The expression levels of Bcl-XL and Bak remained unchanged, whereas the level of Bad was down-regulated. As the ratio between death repressors and death promoters in the Bcl-2 family will determine the sensitivity of cells to apoptotic stimuli, the findings suggest that the changed expression patterns of Bcl-2 family proteins caused by anticancer drugs in liver cancer cells may be involved in chemoresistance. PMID- 10695053 TI - Recombinant and tissue extract thromboplastins for determination of international normalised ratio in over-anticoagulated patients. AB - The international normalised ratio (INR)/international sensitivity index (ISI) system is established for calibration of thromboplastins for laboratory monitoring of oral anticoagulant therapy. The calibration procedure employs patients stabilised on oral anticoagulants, and is therefore validated for patients within the therapeutic range. For practical reasons, the system is used for patients at all levels of therapy, including over-anticoagulated patients with particularly low levels of factors II, VII and X. We studied patients within and above the therapeutic range, using a thromboplastin containing recombinant human tissue factor (Innovin) and two tissue extract thromboplastins. In samples with INRs from 2.0 to 4.0, there was good agreement between results obtained with the three systems (mean INRs within 4% of each other). In patients with INRs > 4.0, results with a human placental extract reagent (Thromborel S) were similar to those obtained with a rabbit brain thromboplastin (IL PT Fib Hs Plus); mean INRs were 6.30 and 6.32 respectively (not significant). Results with Innovin (mean INR: 7.67) were significantly (P < 0.001) greater (on average by 22%) than those obtained with the other two materials. The discrepancy between results with different reagents negatively correlated with factor VII levels. Thus, the lower the factor VII level, the greater was the discrepancy between INRs. Unexpectedly, there was a positive correlation between factor V level and the difference between INRs with different reagents. Thus, the higher the factor V level, the greater was the discrepancy between INRs. The effect of these differences at higher INRs on patient management is unknown, but the recently revised UK guidelines recommend that management of these patients should be influenced by clinical factors, reducing the relative importance of discrepancies between results obtained with different systems. PMID- 10695054 TI - Detection of anti-Yta antibodies using a sensitive and specific enzyme-linked immunosorbent assay. AB - A specific, sensitive and semi-quantitative enzyme-linked immunosorbent assay (ELISA) is described to detect anti-Yta antibodies in human serum. Recombinant acetylcholinesterase (AChE E.C.3.1.1.7) was employed as the coating antigen in the microtitre plate and horseradish peroxidase (HRP)-conjugated specific antibody (IgG) was used as the secondary antibody. The method developed showed excellent sensitivity, detecting a titre > 1 in 600,000 (3.5 ng/mL mouse IgG protein) for mouse monoclonal (mMAb) anti-AChE antibody. No cross-reaction was seen with other common blood group antibodies, confirming the specificity of the method. The recombinant antigen's AChE phenotype was confirmed as Yta, as no reaction was detected with anti-Ytb-positive sera. The ELISA method correlated closely with the established serological grading system used routinely in blood transfusion laboratories. PMID- 10695055 TI - Malignant melanoma: death by image and ignorance, diagnosis by surgical excision and laboratory investigation. AB - Malignant melanoma is now one of the most common cancers in the western world. Across Europe, the rise in annual incidence is 4-7%. The criteria for clinical and histological diagnosis are well established; however, an elaborate range of laboratory investigations can provide valuable supplementary information in difficult cases. In this essay, we highlight the incidence and impact of malignant melanoma worldwide, and outline the range of laboratory investigations that can be performed and are so vital in the diagnosis of problematic cases. PMID- 10695056 TI - Applications of molecular genotyping to immunohaematology. AB - The genes encoding 20 of the 23 blood group systems have been cloned and sequenced. This has allowed the determination of the molecular basis of many blood group antigens and phenotypes. In this review we outline the basis of the genetic code, summarise molecular events that give rise to polymorphic antigens and to blood group phenotypes, and describe how polymerase chain reaction-based techniques can be applied in the clinical setting. PMID- 10695057 TI - Health technology assessment in primary and community care. PMID- 10695058 TI - Supporting doctors, or the beginning of the end for self-regulation? PMID- 10695059 TI - Validity of symptom and clinical measures of asthma severity for primary outpatient assessment of adult asthma. AB - BACKGROUND: Symptom and pulmonary function measures of asthma severity are used for severity classification in practice guidelines. However, there is limited methodological evidence in support of their validity and utility. AIM: To validate initial symptom and forced expiratory volume (FEV1) measures of asthma severity with the subsequent risks of exacerbations resulting in emergency room (ER) visits, hospitalisation, and sickness absence from work. In addition, symptom-based measures of change in asthma severity were also evaluated against the concurrent risks of asthma exacerbations. METHOD: A cohort of 361 adult asthmatic patients in general outpatient clinics was studied. At initial interview, frequencies of asthmatic symptoms and nocturnal exacerbations, FEV1, and a severity score combining these measures, were recorded. At re-interview in the third year, the frequencies of asthma exacerbations resulting in ER visits, hospitalisation, and sickness absence, and a self-assessed global measure of change in severity and serially-assessed change in symptom frequencies, were measured. RESULTS: All individual symptom and FEV1 measures were strongly related to the subsequent risks of ER visits, hospitalisation, and sick absence. A severity score of more than 3 (moderate to severe asthma) and self-assessed change in asthma severity were most strongly and significantly associated with greatly increased risks of all outcomes. Individual symptoms and FEV1 measures alone did not show high sensitivities, but the severity score combining these measures gave much more satisfactory validity. Perhaps not surprisingly, self assessed change in asthma appeared to give the most satisfactory validity. CONCLUSION: These results support the validity and clinical utility of a simple clinical score based on symptom and FEV1 measures, and self-assessed measure of change in severity, for risk classification in contemporary clinical practice guidelines. PMID- 10695060 TI - In-practice evaluation of whole-blood Helicobacter pylori test: its usefulness in detecting peptic ulcer disease. AB - BACKGROUND: Approximately 10% of patients presenting with dyspepsia to the general practitioner have peptic ulcers; the large majority of which are related to infection with Helicobactor pylori. Office-based tests for H. pylori detection are generally validated and evaluated in selected patient groups. AIM: To evaluate the clinical effectiveness of a whole-blood serology test for infection with Helicobacter pylori in detecting peptic ulcer disease (PUD) in daily general practice. METHOD: A descriptive study of 171 primary care dyspepsia patients selected for open-access endoscopy in primary care and aged between 18 and 75 years, in 92 general practices in central, southern, and eastern parts of the Netherlands. H. pylori status was assessed using the BM-test Helicobacter pylori, which is identical to the Helisal test. Dyspepsia severity score was measured using a validated symptom score. Symptom characteristics and probability of relevant disease were assessed by the general practitioner. Endoscopy was carried out in local hospitals. Diagnostic outcome of both endoscopy and H. pylori reference test was supplied by local specialists. The BM-test was evaluated against endoscopic results. RESULTS: A high number (61.8%) of false-negative BM tests resulted in a low sensitivity (95% confidence interval [CI] = 48-75%) for detection of H. pylori infection. Only 12 out of 32 patients with PUD had a positive BM-test, resulting in a positive likelihood ratio (LR) for PUD of 1.41 and a negative LR of 0.85. CONCLUSIONS: This study confirms the relatively poor performance of the BM-test in daily general practice, and shows the limited diagnostic value of H. pylori office-tests for detecting PUD in primary care. The discriminative value of the test result is too small to support either a 'test and-endoscope' of a 'test-and-treat' strategy in general practice. PMID- 10695061 TI - Needs assessment in primary care: general practitioners' perceptions and implications for the future. AB - BACKGROUND: Health needs assessment can guide the appropriate shift to primary care by identifying the most effective and efficient resource allocation to meet the needs of populations. Assessing health care needs will be a continuing challenge for primary care trusts in Scotland (or equivalent groups in other parts of the United Kingdom); however, lessons must be learned from the experience of needs assessment that followed the 'internal market' reforms of the 1990s. AIM: To examine general practitioners' (GPs') awareness and experience of needs assessment, to identify barriers to needs assessment in primary care, and to ascertain how better progress might be made in the future. METHOD: A postal questionnaire survey of 1777 Scottish GPs (a one-in-two sample) was combined with a semistructured interview survey of 'lead' GPs from a random sample of 64 mainland Scottish practices between May and August 1996. RESULTS: Sixty-five per cent (1154) of GPs responded to the questionnaire, of which 54% (965) were completed. Over 73% (47) of interviews were completed. Most GPs were unfamiliar with the concept of needs assessment and there was no evidence that needs assessment had influenced commissioning decisions. Most GPs argued that it was not a 'core' activity and that they lacked training in the relevant skills. While the attitude of the majority was indifferent, cynical, and sometimes hostile, a minority, comprising mostly younger fundholders, was more enthusiastic about needs assessment. CONCLUSION: The motivation and attitude of the majority of GPs present a barrier to needs assessment in primary care. GPs will require more resources and training if they are to undertake this responsibility. Most GPs believe than incentives (financial or organisational) will be necessary. Primary care trusts and equivalent structures should be aware of these attitudes as they seek to establish plans based on estimates of population needs in defined locations. PMID- 10695062 TI - Eating disordered patients: personality, alexithymia, and implications for primary care. AB - BACKGROUND: Eating disorders are becoming more apparent in primary care. Descriptions of character traits related to people with eating disorders are rarely reported in the primary care literature and there is little awareness of the implications of alexithymia--a concept that defines the inability to identify or express emotion. We hypothesised that many individuals with active eating disorders have alexithymic traits and a tendency to somatize their distress. AIM: To analyse the character traits and degree of alexithymia of a selected group of women with active eating disorders and in recovery, and to recommend responses by members of the primary care team that might meet the needs of such individuals. METHOD: Letters were sent to 200 female members of the Eating Disorders Association who had agreed to participate in research. Seventy-nine women volunteered to complete four postal questionnaires. This gave a response rate of 38.5%. Responders were categorised into three groups--anorexic, bulimic, and recovered--using the criteria of the Eating Disorders Inventory (EDI-2). The results of the 16PF5 Personality Inventory (16PF5) and the Toronto Alexithymia Scale (TAS-20) were analysed using one-way analysis of variance (ANOVA) and correlated using Pearson's correlation. A biographical questionnaire was also completed. RESULTS: In all three subgroups, high scores were achieved on the 16PF5 on 'apprehension and social sensitivity', while there were significant differences in the scores for 'privateness': a scale that measures the ability to talk about feelings and confide in others. On the TAS-20, 65% of the anorexic and 83% of the bulimic group scored in the alexithymic range compared with 33% of the recovered group. There was a significant negative correlation between alexithymia and social skills such as 'social and emotional expressivity' on the 16PF5. CONCLUSION: The results of this study emphasise the difference between those with active eating disorders who achieved high scores for privacy, introversion, and alexithymia, and those who have recovered. These character traits give potential helpers an important indication of the areas that can both block and facilitate recovery, and they act as a reminder that the presenting symptoms in eating disorders and other psychosomatic conditions are the outward presentation of internal conflict. It is suggested that effective screening and needs assessment will facilitate a more appropriate and prompt therapeutic response. This may be provided in the primary care setting where appropriate training has occurred. PMID- 10695063 TI - Influenza vaccination in asthma: a primary care experience. AB - BACKGROUND: Despite the recommendation of the Department of Health that patients with asthma receive annual vaccination against influenza, uptake remains unsatisfactory with many patients suspicious that vaccination is harmful. AIM: To examine the effect of influenza vaccination on asthmatic patients typical of a general practice setting. METHOD: A multicentre study with 56 patients participating from 14 practices in England and Scotland. Patients completed peak expiratory flow rate (PEFR) and symptom diaries for two weeks before and two weeks after influenza vaccination. RESULTS: A non-significant fall in baseline PEFR of 10.5 l/min, from an average of 431.5 l/min, was observed after influenza vaccination, representing a 2% change from baseline. A significant increase in night time reliever use of 0.17 puffs per night (P < 0.01) was found. Non significant increases in number of nights per week with sleep disturbed due to asthma, severity of night-time and day-time symptoms, and day-time reliever use were also noted. CONCLUSION: Influenza is an important cause of morbidity and mortality in asthmatics. This study confirms the safety of influenza vaccination in patients with asthma typical of those seen in primary care. General practitioners need not hesitate in recommending this valuable intervention to their asthmatic patients and should consider ways in which uptake can be improved. PMID- 10695064 TI - Do practice-based preventive child health services affect the use of hospitals? A cross-sectional study of hospital use by children in east London. AB - BACKGROUND: Acute paediatric admissions have risen steadily over the past 20 years. During the same period, practice-based child health clinics have increased, although provision is less common in areas of deprivation where hospital use is greatest. AIM: To investigate the contribution of practice-based, preventive child health services to rates of hospital utilisation in children under five years of age. METHOD: A cross-sectional retrospective study examining practice variations in paediatric acute admissions, outpatient referrals, and accident and emergency (A&E) department attendances in the East London and the City Health authority, including all 164 practices in the inner-city boroughs of Hackney, Newham, Tower Hamlets, and the City of London. The main outcome measures were practice-based paediatric hospital attendance rates, for discrete age and sex bands, for the year to 31 March 1996. RESULTS: Hospital use varied with age and sex, with the rates being highest for the youngest children and for boys. The median A&E attendance rate (including reattendances) for boys up to one year of age was 897 per thousand children per practice. In east London, 62% of practices are registered for child health surveillance and 71% provide a child health clinic. Practice approval for child health surveillance, and the provision of child health clinics, did not account for differences between practices in hospital use, but proportionally greater health visiting hours were significantly related to lower rates of emergency hospital admission by young children. Multivariate analyses revealed that up to 23% of the variation between practice admission rates could be explained by health visiting hours. CONCLUSIONS: We found significant associations between the amount of health visiting time available to the practice population and rates of acute admission and outpatient referral among children up to five years of age. These findings suggest that increasing health visitor provision could contribute to lower paediatric emergency admission and outpatient referral rates. A small change would have a significant effect, particularly among the youngest children, given that during the study year 10,000 children under two years of age in east London were either admitted or referred to hospital. PMID- 10695065 TI - Management confidence and decisions to refer to hospital of GP registrars and their trainers working out-of-hours. AB - BACKGROUND: There is concern about the educational impact and possible stress on registrars of new out-of-hours co-operatives. AIM: To compare the confidence in managing out-of-hours problems of registrars in traditional on-call rotas and co operatives with that of their trainers. To determine how frequently registrars discussed problems out-of-hours with their trainers, and to compare the referral pattern of registrars with their trainers out-of-hours. METHOD: Analysis of log diaries of out-of-hours experiences kept by registrars and trainers over two, two month periods in winter and summer. RESULTS: Thirty registrars (out of a possible 51) and 34 (out of a possible 52) trainers took part in the winter, and 18 registrars and 29 trainers in the summer. Registrars were confident in their management, and their confidence increased over the year (59% versus 72% difference = 12%, 95% CI = 6% to 20% for very confident). Registrars varied in their discussion of problems with trainers. When 'a little worried' they discussed their management 30 out of 53 times (57%); if 'very confident', 36 out of 576 times (6%). Registrars during the summer segment of the study referred more frequently to hospital than trainers (20% versus 10% difference = 10%, 95% CI = 3% to 17%. Registrars in traditional rotas recorded a slightly higher but statistically insignificant level of confidence in their management of problems than those registrars in cooperatives. CONCLUSIONS: While many registrars are confident in their work and are using their trainer for information appropriately, some are not. Registrars may be referring to hospital at a much higher rate than their trainers. More research is required to confirm and further explore these findings. PMID- 10695066 TI - Doctor-staffed ambulance helicopters: to what extent can the general practitioner replace the anaesthesiologist? AB - During two years, a rural ambulance helicopter programme saved 41 patients' lives. In 29 of these patients, the decisive medical interventions were carried out by the flight anaesthesiologist before reaching the hospital. We asked an expert panel to assess whether these interventions could have been carried out by a general practitioner (GP). This was the case for 17 (59%) of the 29 patients, while more advances skills, equipment or drugs were needed for 11 (38%). Among these 11, three patients would probably have died without the interventions. We conclude that GPs can manage a majority of life saving missions for a rural ambulance helicopter programme, but the lack of a flight anaesthesiologist may imply substantial health losses for a few patients. PMID- 10695067 TI - Can dietary assessment in general practice target patients with unhealthy diets? AB - Diet is important in the aetiology and management of many conditions in primary care. Although valid dietary assessment is required for both clinical work and research, no dietary assessment instruments have been validated among patients seen in primary care. A range of simple self-completion dietary assessment questionnaires and established research instruments were compared with an accepted reference standard, a seven-day weighed record, in 111 subjects assessed in a practice nurse-run treatment room. Simple self-completion tools based on food groups and portion sizes perform as well (likelihood ratios for a positive test = 2 to 3) as much more time-consuming instruments. The error in using such instruments is comparable with the error of the standard itself. There is little justification for using time-consuming dietary assessment questionnaires, since simple tools are accurate enough to be clinically useful--to allow practice nurses to target patients for counselling and waste less time on inappropriate counselling--and also useful for research. PMID- 10695068 TI - To what extent are practices 'paperless' and what are the constraints to them becoming more so? AB - A questionnaire was sent to all practice managers in Wessex in June 1997 to assess to what extent practices had stopped relying on paper records in the consultation. Practices that solely used computer records in the consultation ('noteless') did not necessarily consider themselves 'paperless'. Following a recent declaration that all practices should be using an Electronic Health Record by March 2005, the obstacles to this move were investigated and some differences in the way the 'noteless' and 'paperless' groups used computers were identified. PMID- 10695069 TI - Can methadone maintenance for heroin-dependent patients retained in general practice reduce criminal conviction rates and time spent in prison? AB - A retrospective analysis was made of the criminal records of 57 patients successfully retained in methadone maintenance at two general practices in Sheffield. Their criminal conviction rates and time spent in prison per year were compared for the periods before and after the start of their methadone programme. Overall, patients retained on methadone programmes in the general practices studied had significantly fewer convictions and cautions, and spent significantly less time in prison than they had before the start of treatment. PMID- 10695070 TI - Diagnosis of patients with chronic heart failure in primary care: usefulness of history, examination, and investigations. AB - Chronic heart failure is a common clinical syndrome that may have different causes. Its incidence and prevalence are predicted to rise substantially over the next 10 years. There are therefore major consequences for resource provision, especially in primary care, where most patients are managed. Chronic heart failure is a serious condition with high morbidity and mortality. There is good evidence to show that treatment with angiotensin-converting enzyme (ACE) inhibitors in patients with left ventricular systolic dysfunction improves symptoms and signs, slows progression of heart failure, reduces hospitalisation rates, and improves survival. Despite this evidence, primary care studies show that patients with heart failure are incorrectly diagnosed and inadequately treated. Most patients present in general practice, and because effective treatment relies on a correct diagnosis, this is a key step in the appropriate management of heart failure. The aim of this paper is to review the evidence about the usefulness of signs, symptoms, and investigations in diagnosing heart failure in primary care. To identify relevant studies for this review, four strategies were used: a MEDLINE search from 1993 to January 1998 using the diagnosis search filter; a MEDLINE search from 1993 to January 1998 using the guideline search filter to locate published heart failure guidelines; a search for review articles in the Cochrane Library; and a check of references in the studies identified. The search terms included MeSH terms and the keywords 'heart failure' and 'diagnosis'. All searches were limited to humans and English language articles. Studies were included in this review on the basis of quality and relevance to primary care. The review shows that symptoms and signs are important because they alert clinicians to the possibility of heart failure as a diagnosis. However, they are not sufficiently specific for confirming left ventricular systolic dysfunction. From the evidence available, a patient with suspected heart failure must have objective tests to confirm the diagnosis. These should include an electrocardiogram and, ideally, an echocardiogram. Further research is also needed on the usefulness of signs and symptoms in primary care, as most studies of heart failure have been conducted in secondary care. PMID- 10695072 TI - Supporting practice-based audit. PMID- 10695071 TI - What do GPs need to know? The use of knowledge in general practice consultations. AB - In the course of the consultation in primary care, the general practitioner integrates knowledge of different types that are drawn from different sources. As a consequence of the way practitioners develop expertise, this use of knowledge is often hidden from the conscious mind of the practitioner and often hidden from direct observation. On the other hand, understanding of this use of knowledge is crucial to several necessary developments of the profession of general practice. A method involving collaboration between researcher and practitioner sheds new light on this knowledge-in-use. PMID- 10695073 TI - The reform of GP training. PMID- 10695074 TI - Providing health care for the homeless. PMID- 10695075 TI - Recruitment and retention of GPs in the UK. PMID- 10695076 TI - Epidemiology of intensive care medicine: supply versus demand. AB - Developments in hospital medicine combined with social and demographic changes are likely to increase the need for intensive care services at a time when cost containment and cost-efficacy are the main items on the political agenda. This will exaggerate the supply-demand outcome mismatch unless the problem is approached in a constructive manner by clinicians, managers and politicians. More resources will be required for intensive care, but these must be better targeted and more efficiently employed. Opportunities for prevention should be explored, with intensive care being given a pro-active rather than a re-active role. Intensive care clinicians should understand that this expanded role cannot be achieved if they are willing only to accept responsibility for patient care after the patient has been admitted to the ICU. Clinicians and managers should develop methods for linking the various disciplines which contribute to emergency care, to form an acute care framework within the hospital. Research into the factors which determine risk of critical illness should be combined with enhanced medical and nursing training in intensive care, accompanied by an expansion in resources for intermediate and high dependency care in countries like the UK where there is clear evidence of rationing. PMID- 10695077 TI - Inflammatory cell activation in sepsis. AB - The body relies for protection on an effective inflammatory response. To sustain an armoury of inflammatory cells in a state of permanent activation would be impossible and a system whereby such cells can be rapidly activated is, therefore, employed. Upon transition from the resting to activated state inflammatory cells perform multiple defensive functions and are then removed, limiting the duration of inflammation. Neutrophils are the major circulating inflammatory cells but macrophages exert a more powerful regulatory effect. If the inflammatory response is inadequate there is a risk of overwhelming sepsis. By contrast, an unregulated response can lead to systemic inflammation and consequent multiple organ damage. This review focuses on the mechanisms whereby inflammatory cells are activated, how the regulatory system may misfunction and how it may in the future be manipulated to therapeutic advantage. PMID- 10695078 TI - The role of the endothelium in modulating vascular control in sepsis and related conditions. AB - The majority of deaths amongst critically ill patients requiring intensive care are attributable to sepsis and its sequelae: septic shock, the systemic inflammatory response syndrome (SIRS) and the acute respiratory distress syndrome (ARDS). Patients within the ICU who develop these conditions and fail to survive succumb to multiple organ dysfunction syndrome (MODS). ARDS is considered to be the pulmonary component of MODS and is characterized by pulmonary hypertension, often in the setting of systemic hypotension. Endothelial cells, normally responsible for modulating vascular tone, becomes dysfunctional in sepsis. Pro thrombotic, pro-inflammatory and vasoactive mediators are released including nitric oxide (NO), endothelins (ETs) and products of cyclo-oxygenase metabolism. It is probably the disordered production of these mediators in vascular beds that results in MODS. This review highlights recent research in this area with particular emphasis on possible therapeutic options. PMID- 10695079 TI - Redox imbalance in the critically ill. AB - The majority of deaths amongst critically ill patients requiring intensive care are attributable to sepsis and its sequelae: septic shock, the systemic inflammatory response syndrome (SIRS) and the acute respiratory distress syndrome (ARDS). Clinically, sepsis/SIRS and ARDS are characterised by disordered vascular control, manifest as systemic hypotension and peripheral vasodilation refractory to intravascular volume resuscitation and vasopressor therapy; and pulmonary hypertension. Experimental and clinical evidence demonstrates that these patients suffer from severe oxidative stress. Thus, our own and other groups have shown that the vascular pathology of sepsis/SIRS and ARDS is initiated through the uncontrolled production of reactive oxygen (ROS) and reactive nitrogen species (RNS) which modulate inflammatory cell adhesion and cause direct injury to endothelium (Fig. 1). PMID- 10695080 TI - Sedation and analgesia. AB - Sedation is a process of soothing. The concept of the ideal level of sedation is controversial and has changed over the last decade. A shift from deep sedation, often enhanced by muscle relaxants that completely detaches the patient from their environment, to light sedation rendering the patient sleepy but easily arousable has been widely accepted. This change in attitude has been brought about by sophisticated modes of ventilation allowing the ventilator to synchronize with the patient's own breathing pattern. In addition, the increasingly recognised adverse effects of over-sedation have contributed to the reduction in the depth of sedation. PMID- 10695081 TI - The oxygen trail: tissue oxygenation. AB - Aerobic cellular respiration depends on the efficient supply of oxygen and substrate to the mitochondria. There is an oxygen cascade from the environment to the subcellular environment. Efficient oxygen delivery depends on the coordinated interaction between the respiratory and circulatory systems. The circulation at both macro- and microvascular levels is under the control of humoral and neural factors. There is local autoregulation of flow at the tissue level by metabolic factors which reflect the energy state of the tissues. The response to hypoxia involves the activation of cytokines and genetically controlled factors which maximise capillary perfusion and haemoglobin concentration, and regulate cell metabolism. The formation of reactive oxygen species under such conditions has a detrimental effect on the mitochondria with respiratory chain dysfunction, increased permeability transition, and cell death. This review aims to explore the mechanisms by which the body attempts to maintain tissue oxygen levels at conditions optimal for cell survival. PMID- 10695082 TI - The oxygen trail: measurement. AB - Tissue hypoxia may be defined as abnormal oxygen utilization such that cells are experiencing anaerobic metabolism. Tissue hypoxia can be defined biochemically by low levels of ATP, high levels of NADH, or decreased oxidized cytochrome aa3. It is possible to measure these biochemical markers in the laboratory setting with, for example, nuclear magnetic resonance spectroscopy. However, this is not as yet a clinical option. There is no 'gold standard' for the diagnosis of clinical hypoxia. We can detect the gross consequences of tissue hypoxia, such as organ dysfunction and metabolic markers of anaerobic metabolism (e.g. lactic acidosis). We have also become familiar with the measurement of both global and regional oxygen dispatch and consumption. However, organ dysfunction and metabolic acidosis consistent with established tissue hypoxia commonly exists in the presence of normal and even supra normal global measures of oxygen dispatch and consumption. Therefore, we should ideally make measurements at the end of the oxygen trail, i.e. cellular oxygen delivery and effective utilization. PMID- 10695083 TI - The oxygen trail: the goal. AB - Over the last 10 years, there have been great advances in knowledge concerning changes in tissue perfusion and its prognostic implications. Has this translated into improved patient management? We review the clinical trials that have deliberately increased tissue oxygen delivery by increasing cardiac output. We have divided the studies into those that intervene early or those that intervene late in the course of a patient's illness. Although there are methodological problems limiting interpretation of the results, we show a combined odds ratio of a reduction in mortality for the early studies but not for the late studies. We conclude that a treatment policy whereby oxygen delivery is deliberately increased improves patient outcome if it is initiated early, prior to the onset of organ failure. PMID- 10695084 TI - Ventilatory support in the acute respiratory distress syndrome. AB - Ventilatory support in the acute respiratory distress syndrome (ARDS) has undergone considerable transformation in the 1990s. Current approaches include lung protective techniques which, while attempting to recruit and maintain lung volume, limit the shear stresses associated with ventilation by avoiding both alveolar overdistension and cyclical end-expiratory collapse. In addition, gas exchange targets have been liberalized and ventilatory conduct is much more tailored to individual pulmonary mechanics. Assessment of the inspiratory volume pressure (V-P) curve provides information which can direct ventilator settings. Recent information from clinical trials has provided new insights into appropriate ventilatory modification and set the foundation for future clinical investigations. PMID- 10695085 TI - Non-ventilatory treatment of acute hypoxic respiratory failure. AB - Severe acute hypoxic respiratory failure is uncommon but often fatal. Standard treatment involves high inspired oxygen concentrations, mechanical ventilation and positive end-expiratory pressure. Many other interventions have been used in parallel with conventional treatment or as rescue therapy when it fails, including extracorporeal gas exchange, prone positioning, inhaled vasodilators, exogenous surfactants and drugs which modify the inflammatory process. Nearly all these treatments improve arterial oxygenation or markers of lung injury. However, the relationship between oxygenation and survival in acute hypoxaemic respiratory failure is not established, so improved oxygenation cannot be used as a surrogate for survival. Randomised controlled trials are, therefore, needed to assess the effects of these treatments on mortality. In such trials, extracorporeal oxygenation and extracorporeal carbon dioxide elimination, surfactant, early methylprednisolone, and prostaglandin E1 offer no survival advantage over conventional therapy. Prophylactic ketoconazole and pentoxifylline appear to improve mortality in small studies in surgical and oncology patients respectively, and methylprednisolone improves mortality and morbidity in unresolving disease. PMID- 10695086 TI - Manipulating the metabolic response to injury. AB - In this short review we will concentrate on just one of the features of the metabolic response to injury (classified as accidental trauma, injury or sepsis) which are collectively known as the 'flow' phase. These include an increase in energy expenditure (hypermetabolism), changes in substrate utilisation (insulin resistance) and the focus of this chapter muscle wasting or catabolism. It is recognised that the three features are interrelated, for example insulin is believed to be an important factor in controlling amino acid flux in skeletal muscle and increasing environmental temperature which may reduce flow phase hypermetabolism has been shown to reduce postoperative nitrogen excretion (a marker of protein catabolism). However, we will concentrate on muscle wasting and refer the reader to other reviews on insulin resistance and metabolic rate. PMID- 10695087 TI - The gastrointestinal tract as a barrier in sepsis. AB - The gastrointestinal tract is an organ of digestion and absorption which is metabolically active and has specific nutrient requirements. In health, it has an additional function as a major barrier, protecting the body from harmful intraluminal pathogens and large antigenic molecules. In disease states, such as sepsis when the mucosal barrier is compromised, micro-organisms and their toxic products gain access to the portal and systemic circulations producing deleterious effects. Under these circumstances, systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) develop leading to deterioration and death of the patient in the intensive care unit. Therapeutic strategies for such patients in the intensive care unit aim to support general immune function and maintain the structure and function of the gastrointestinal tract. For these therapies to be successful, the underlying septic or necrotic focus must be ablated using appropriate surgical or other invasive techniques. PMID- 10695088 TI - Adjunctive therapy in sepsis: a critical analysis of the clinical trial programme. AB - Despite intensive efforts, the development of novel drugs for the treatment of sepsis has proved to be extremely difficult. A large number of clinical trials have ended in failure. A critical analysis of this record suggests that there is no single reason for these problems. Rather, the explanation lies in part with unexpected failures in the drugs themselves, and in part with the difficulties of trial design in this particular group of patients. In future, trials in this area are likely to be more highly focused, with even stricter protocol definitions to try and ensure a homogeneous patient population. PMID- 10695089 TI - Cerebral protection in severe brain injury: physiological determinants of outcome and their optimisation. AB - The primary role of intensive care in acute head injury lies in the prevention, detection and reversal of secondary neuronal injury. The maintenance of optimal systemic and cerebrovascular physiology can substantially contribute to these aims. There is, however, a role for novel neuroprotective interventions, many of which are currently under investigation. PMID- 10695090 TI - Antibacterial resistance in the intensive care unit: mechanisms and management. AB - The incidence of multiple antimicrobial resistance of bacteria which cause infections in the intensive care unit is increasing. These include methicillin resistant Staphylococcus aureus and vancomycin-resistant enterococci, penicillin resistant Streptococcus pneumoniae and cephalosporin and quinolone resistant coliforms. More recently, pan antibiotic resistant coliforms, including carbapenems, have emerged. The rapidity of emergence of these multiple antibiotic resistant organisms is not being reflected by the same rate of development of new antimicrobial agents. It is, therefore, conceivable that patients with serious infections will soon no longer be treatable with currently available antimicrobials. Strict management of antibiotic policies and surveillance programmes for multiple resistant organisms, together with infection control procedures, need to be implemented and continuously audited. As intensive care units provide a nidus of infection for other areas within hospitals, this is critically important for prevention of further spread and selection of these resistant bacteria. PMID- 10695091 TI - Red blood cell substitutes: fluorocarbon emulsions and haemoglobin solutions. AB - The problems posed by transfusion of homologous blood have led to the development of substances able to replace the gas transporting properties of blood. Perfluorocarbons (PFCs) emulsions and modified haemoglobin (Hb) solutions have been developed for this goal and are now tested in clinical assays. PFCs are synthetic fluorinated hydrocarbons, capable of dissolving large quantities of oxygen (O2; without binding) at high inspired concentrations of O2, and of delivering this O2 to the tissues. They are administered as emulsions containing particles with a diameter of approximately 0.2 micron, capable of entering the microcirculation. They are eliminated unchanged by the lungs within several days. Fluosol-DA 20% was the first PFC emulsion used in clinical practice. Currently, Oxygent, a second generation PFC emulsion, is being evaluated in clinical studies. The PFCs are not blood substitutes, but rather a means to ensure tissue oxygenation during extreme haemodilution. Solutions of free Hb do not have the antigenic characteristics of the blood groups, and do not require compatibility testing. They are fully saturated with O2 at ambient FiO2. The Hbs used are derived from either human or bovine sources, or via recombinant DNA technology. In order to maintain satisfactory intravascular half-life and O2 affinity, the Hb molecules are modified by adding internal crosslinks, by polymerization, and/or by encapsulation. After promising animal studies, several of these modified Hb solutions are now being studied in Phase III clinical trials. Among them, diaspirin cross-linked haemoglobin (DCLHb) has been used in cardiac and orthopaedic surgery, and for resuscitation of traffic accident victims. The initial results of multicentre trials are now being analysed. PMID- 10695092 TI - Medicines cheaper over the counter. May 1999. PMID- 10695093 TI - Erectile dysfunction--the right to be treated. PMID- 10695094 TI - Efficacy of twice-daily amoxycillin/clavulanate in lower respiratory tract infections. AB - In this double-blind, double-dummy study, 324 patients with clinical evidence of community-acquired pneumonia (CAP) or an acute exacerbation of chronic bronchitis were randomly assigned to receive 10 days' treatment with either amoxycillin/clavulanate 875/125 mg twice daily or amoxycillin/clavulanate 500/125 mg three times daily. At the end of therapy, clinical success rates were 92.4% for the twice daily regimen and 94.2% for the three times daily regimen. There was no statistically significant difference between treatments (p = 0.647) and the 95% confidence interval around the treatment difference indicated that the two treatments were equivalent. Treatment equivalence was also confirmed at follow-up, four weeks after the end of treatment. Both regimens were well tolerated. In conclusion, amoxycillin/clavulanate 875/125 mg twice daily is as effective as amoxycillin/clavulanate 500/125 mg three times daily for the treatment of community-acquired lower respiratory tract infections and could improve patient compliance. PMID- 10695095 TI - Switch to non-CFC inhaled corticosteroids: a comparative efficacy study of HFA BDP and CFC-BDP metered-dose inhalers. AB - Inhaled corticosteroids, such as beclomethasone dipropionate (BDP), recommended for the treatment of persistent, mild, moderate, or severe asthma, have traditionally been administered via chlorofluorocarbon (CFC) propellant. The imminent phasing out of CFCs from pharmaceutical preparations due to the Montreal Protocol means patients will have to switch to a CFC-free alternative. One such preparation is hydrofluoroalkane-BDP (HFA-BDP), a press-and-breathe metered-dose inhaler. This 8-week, open-label, multicentre study assessed asthma control in patients switching from CFC-BDP to HFA-BDP (QVAR). Patients with asthma, stabilised on 400-1600 micrograms/day CFC-BDP, were randomised to HFA-BDP (354 patients; 75%) at half their current daily dose of CFC-BDP, or to CFC-BDP (119 patients; 25%). HFA-BDP efficacy was found to be equivalent to that of CFC-BDP in that no statistically significant difference was observed between the two groups in the mean change from baseline in a.m. PEF (95% CI within +/- 11 l/min). No statistically significant differences were observed between the two groups for increased asthma symptoms or acute asthma episodes. We conclude that asthma control was maintained over 8 weeks, with few asthma exacerbations, in patients switching from previous CFC-BDP therapy to HFA-BDP at half the dose. PMID- 10695096 TI - A comparison of the effects of two modified release preparations of nifedipine nifedipine retard 10 mg twice daily and nifedipine GITS 20 mg once daily--in the treatment of mild to moderate hypertension. AB - A multicentre, randomised, double-blind, cross-over comparison of nifedipine GITS 20 mg once daily and nifedipine retard 10 mg twice daily in 49 patients with mild to moderate hypertension was conducted. Both treatments resulted in clinically significant trough blood pressure reductions (nifedipine GITS -10.1/-8.9 mmHg, nifedipine retard -7.5/-8.2 mmHg). The study demonstrated that nifedipine GITS was 'at least equivalent' to nifedipine retard in the reduction of trough diastolic blood pressure (one-sided lower 95% confidence limit, -1.2 mmHg). The overall incidence of adverse events (nifedipine GITS 25.5%, nifedipine retard 34.0%), as well as the incidences of headache (nifedipine GITS 8.5%, nifedipine retard 12.8%) and peripheral oedema (nifedipine GITS 2.1%, nifedipine retard 8.5%), was higher with nifedipine retard compared to nifedipine GITS. Nifedipine GITS 20 mg once daily is 'at least equivalent' to nifedipine retard 10 mg twice daily in patients with mild to moderate hypertension, as well as being better tolerated. PMID- 10695097 TI - A comparison of nimesulide beta cyclodextrin and nimesulide in postoperative dental pain. AB - The aim of this study was to assess the efficacy and tolerability of single doses of nimesulide beta cyclodextrin compared with nimesulide in patients with dental pain following surgical procedures. This was a randomised, double-blind, between patient, multicentre study involving 148 outpatients suffering from moderate to severe pain, who received single doses of either 400 mg nimesulide beta cyclodextrin or 100 mg nimesulide. The principal criterion for efficacy was pain intensity assessed on a visual analogue scale (VAS) 15 minutes after drug intake. Pain intensity was further evaluated 30, 45, 90, 120, 180, 240 and 360 minutes after dosing. Pain relief was evaluated at the same time points by means of a categorical scale. The time point of first pain relief, the use of rescue medication and the global evaluation of efficacy were also recorded. The reduction in pain intensity was significantly more pronounced in the nimesulide beta cyclodextrin group at 15, 30, 45 and 60 minutes (p < 0.01). Pain relief was significantly greater (p < 0.05) and more rapid with nimesulide beta cyclodextrin. In the patient overall assessment of efficacy, nimesulide beta cyclodextrin and nimesulide were rated excellent or good by 95% and 92% respectively; only one patient in the nimesulide beta cyclodextrin group needed rescue medication. Both study drugs were effective and well tolerated in the treatment of acute dental pain, with nimesulide beta cyclodextrin showing a faster onset of analgesic action. PMID- 10695098 TI - A randomised, double-blind study of itraconazole versus placebo in the treatment and prevention of oral or oesophageal candidosis in patients with HIV infection. AB - HIV-infected patients presenting with oral or oesophageal candidosis were randomised to four weeks treatment with itraconazole 200 mg, followed by itraconazole or matching placebo for a prophylaxis phase of 24 weeks. Clinical and mycological evidence of candidosis infection was assessed on a four-weekly basis. Seventy patients were enrolled, of whom 50 completed 28 days of itraconazole therapy; 74% (37 patients) were clinically cured and 40% were also mycologically cured. Fifteen patients were withdrawn for a variety of reasons including non-compliance, adverse events and the use of excluded medications. Forty-four patients were enrolled in the prophylactic phase. There were significantly more relapses of candidosis, and time to candidosis was significantly shorter in the placebo group than in the itraconazole treated group (p = 0.0001). Itraconazole 200 mg daily is effective and well tolerated for the treatment and prevention of oral and oesophageal candidosis in HIV-infected patients. PMID- 10695099 TI - Anaesthetic assessment and management of cardiac patients for non-cardiac surgery. Part 2: Management. AB - In an earlier article in this journal (June 1999) we discussed the risk that the presence of cardiac disease poses to patients undergoing non-cardiac surgery. We outlined factors in the patient's medical history, examination findings and the value of various tests in arriving at an overall assessment of risk for any given patient. In this article we concentrate on the management of these patients as they undergo surgery itself. We shall consider what measures may usefully be employed in order to minimise the risk of an adverse cardiac event occurring in the perioperative period. PMID- 10695100 TI - Polycystic ovarian syndrome: is community care appropriate? AB - In recent years the complexities and heterogeneity of polycystic ovarian syndrome (PCOS) have been recognised. Most PCOS sufferers with amenorrhoea, menstrual dysfunction, hirsutism, acne and infertility can be evaluated and safely managed in primary care. It is prudent to remember that all women with PCOS are at risk of insulin resistance and the associated abnormalities of the insulin resistance syndrome--dyslipidaemia, hypertension, coronary artery disease and diabetes mellitus. PMID- 10695101 TI - Erectile dysfunction and cardiovascular disease. AB - Patients with cardiovascular disease are at increased risk of developing erectile dysfunction (ED). This may be a consequence of atherosclerosis of the penile arteries, a reduced cardiac output, or a side-effect of drugs used to reduce cardiovascular risk factors (particularly beta-blockers, thiazide diuretics and, occasionally, lipid-lowering drugs). ED is a distressing condition, which often diminishes the patient's self-esteem, with the potential for damage to his psychological health and his relationship with his partner and family. When treating ED, the underlying aetiology should be established by careful examination and consideration of medical history and concurrent medication. Until recently, pharmacological treatment options involved intracavernous injections (alprostadil or moxisylyte) or intraurethral alprostadil. These treatments are often inconvenient and not well accepted by the patient. The recent introduction of oral sildenafil promises to revolutionise the treatment of ED. In double blind, placebo-controlled trials in patients with ED, sildenafil improved erectile function and quality of life and was well tolerated. ED is a clinically important complication of cardiovascular disease and should be asked about and treated accordingly. It is important that effective treatments, including sildenafil, should be available for treating patients with cardiovascular disease and ED. PMID- 10695102 TI - Treatment guidelines for hypercholesterolaemia: time to consider soluble fibre. AB - Lowering raised serum cholesterol levels is firmly established as an effective intervention for reducing the mortality and morbidity due to coronary heart disease (CHD). Recent European and British guidelines for the management of hypercholesterolaemia recommend dietary modification as first-line therapy. However, dietary measures alone do not significantly reduce cholesterol levels. There is ample evidence that soluble fibre, such as ispaghula, lowers cholesterol and could therefore potentially reduce the risk of CHD. As CHD management costs spiral upwards, prescribing lipid-lowering drugs for all indicated patients is not sustainable within current NHS resources. It is suggested that treatment guidelines be revised to include a soluble fibre product, such as ispaghula, as an adjunct to diet for patients where diet alone has failed and where lifelong therapy with lipid-lowering drugs is inappropriate. PMID- 10695103 TI - The management of young dyspeptic patients in the era of Helicobacter pylori. AB - The management of young dyspeptic patients remains controversial in the modern Helicobacter pylori era. The use of non-invasive screening for H. pylori in one proposed strategy has demonstrated a substantial reduction in the endoscopy workload by excluding H. pylori negative patients under the age of 45 years with uncomplicated dyspepsia. An alternative screening strategy proposes a 'test and treat' approach, with H. pylori positive patients proceeding directly to an empirical course of eradication therapy. Ednoscopy would be reserved for patients who failed to respond symptomatically or who were H. pylori negative on intial screening. At present there are few data available from clinical studies of putting the 'test and treat' policy into practice. Although there is likely to be a role for screening young dyspeptic patients for H. pylori in primary care, subsequent management requires well-planned studies in order to assess the benefits of any particular strategy. PMID- 10695104 TI - Risk factor modification extends the benefit of coronary artery revascularisation procedures. AB - The management of coronary artery disease has undergone major changes in the past three decades. Early medical preventive strategies rapidly gave way to surgical intervention by the end of the 1960s; 10 years later this was supplemented by the introduction of angioplasty. Both of these approaches offered rapid symptomatic relief but were dogged by a predisposition to restenosis. While technical improvements have helped reduce this problem, there is a growing perception that the imbalance between the enthusiastic adoption of surgical intervention and the jaundiced neglect of secondary medical prevention needs to be redressed. This can be achieved without stepping back in time, for, since the 1960s, agents far more effective in the struggle against ischaemic vascular disease have proven their worth in the clinical arena. Graft protection following bypass surgery is vital, but equally important is the need to target the metabolic derangement which caused the occlusion in the first place. This article reviews medical strategies designed to add life to coronary artery revascularisation procedures. PMID- 10695105 TI - Therapeutic approaches to the management of common baldness. AB - Common baldness or androgenetic alopecia is a universal problem, having affected both sexes of all races to different extents for as long as mankind has existed. The progressive androgen dependent hair loss follows specific patterns and is a physiological process, which may take on disease quality due to psychosocial compontents. These should be taken into consideration when discussing the several treatment options available, particularly as a cure cannot be offered. Cosmetic measures range from back-combing over perms to hairpieces and wigs. Medical therapies include systemic antiandrogens and topical minoxidil; surgical options comprise follicular transplantation, scalp reduction and rotation. Before starting treatment, however, careful consideration should be given to the differential diagnosis which includes drug-induced hair thinning, anagen effluvium, diffuse alopecia areata, metabolic disturbances, nutritional deficiencies and acute as well as chronic telogen effluvium. PMID- 10695106 TI - The stigma of eating disorders. AB - Those who suffer from eating disorders often carry the added burden of stigmatizing attitudes from the lay public and the medical profession. These attitudes not only restrict the opportunities for effective treatment but also confer additional handicaps. To some extent, stigmatizing beliefs are based on partial truths about these disorders, namely their dangerousness, their sometimes poor response to treatment, the sufferers' part in their maintenance, and difficulties in communication. This review explores the truth of these beliefs and suggests ways in which a more positive approach to the management of eating disorders might help to reduce the stigma. This includes empathy with the patient's predicament and an approach to treatment in which the patient's wishes are paramount. PMID- 10695107 TI - Unusual presentation of dissecting aortic aneurysm. AB - Dissecting aneurysms generally cause radiating back pain, chest pain, or symptoms caused by aortic insufficiency. Presentation solely with abdominal pain is rare. We report on a patient with dissecting thoracic aortic aneurysm who presented solely with abdominal pain. The possibility of intrathoracic disease must be considered in every patient with abdominal pain, especially if the pain is in the upper part of the abdomen. PMID- 10695108 TI - Mucocoeles of the maxillary sinus. AB - Mucocoeles of the paranasal sinuses most commonly occur in the frontal or anterior ethmoidal sinuses. We report two rare cases of mucocoele of the maxillary sinus and describe the presentation, investigations and treatment. A review of the literature on this rare clinical entity is included, with specific reference to diagnostic imaging to distinguish mucocoeles from neoplastic sinus disease. PMID- 10695109 TI - Asymptomatic pituitary apoplexy after aortocoronary bypass surgery. AB - Pituitary apoplexy usually presents with acute neuro-ophthalmological complications that require urgent neurosurgical intervention. We present a case of pituitary apoplexy following aortocoronary bypass surgery that was asymptomatic until the patient presented with features of hormonal deficiency three months later. Only one case of pituitary apoplexy has been described in the literature following cardiac surgery that did not require operative intervention. We discuss the aetiology of pituitary apoplexy and the possible mechanisms for such an event after cardiac surgery. Although this is rare, any unusual feature after operation such as lethargy or erectile dysfunction should remind us of hypopituitarism. PMID- 10695110 TI - The use of recombinant human G-CSF in the treatment of propylthiouracil-induced agranulocytosis. AB - A 43-year-old female patient with Basedow-Graves' disease developed agranulocytosis in the eighth month of propylthiouracil therapy. After discontinuing the drug, a broad spectrum antibiotic regimen plus recombinant human granulocyte colony-stimulating factor (G-CSF), a human haematopoietic growth factor, were started. Her granulocyte count returned to normal with the second dose of G-CSF, and ulcerating pharyngitis improved rapidly. We think that in patients with propylthiouracil-induced agranulocytosis, G-CSF will reduce the risk and severity of infection, and should be accepted as a part of the standard therapy. PMID- 10695111 TI - An unusual cause of falls in an elderly patient. AB - Falls are common in the elderly, often causing considerable morbidity and mortality. Prevention is therefore important and is based on determining the cause. We present an elderly patient who had multiple falls during the day due to recurrent daytime sleep episodes, an entity we believe has not previously been reported. PMID- 10695112 TI - Delayed diagnosis of an enteric duplication cyst. AB - Enteric duplication cysts are rare and often prove extremely difficult to diagnose. We report a case in which the definitive diagnosis was made 40 years after first presentation and following five exploratory laparotomies. A review of the literature is presented, with discussion regarding the presenting features, potential complications, radiological investigations and surgical treatment of this uncommon clinical entity. PMID- 10695113 TI - On the chronotherapy of asthma. PMID- 10695114 TI - Quality of life in Parkinson's disease. PMID- 10695115 TI - The Institute of Naval Medicine. PMID- 10695116 TI - Nursing: an all graduate profession? PMID- 10695117 TI - Nagorno Karabakh. PMID- 10695118 TI - Peutz-Jeghers syndrome--a case report. AB - A case of Peutz-Jeghers Syndrome presenting acutely with ileo-ileal intussusception is described. Management issues including follow up protocols are reviewed with reference to the current literature. PMID- 10695119 TI - Operational telemedicine. AB - A simple, effective digital camera and E-mail-based telemedicine system has been developed using commercially available equipment. Initial trials were successful and this system is now deployed in several operational units. A retrospective audit has shown it to be of value in 50% of referrals. PMID- 10695120 TI - EMIS: the story so far. PMID- 10695121 TI - QARNNS at sea--HMS Roebuck. AB - Despite an initial problem of seasickness I thoroughly enjoyed my five weeks at sea on board HMS Roebuck. Having to make medical decisions can be somewhat daunting. However, you always have back up radio support if required. Although only at sea for a short time, my experience has given me a good insight into life at sea especially with both men and women serving on the same ship. With no experience of serving on board larger ships I can recommend that, if possible further QARNNS ratings are given the opportunity to sample life on board HMS Roebuck and that their presence on board would be appreciated by the ship's company, especially with their forthcoming East Coast Surveys. All that remains to say is to thank the Captain and the rest of the ship's company for making me feel so welcome and that the terms 'For Exercise, for exercise, for exercise and Machinery Breakdown' will be sorely missed.....not! ('fair one!' as the Buffer would say). PMID- 10695122 TI - Body composition and fitness of Royal Naval officer cadets. PMID- 10695123 TI - Monckton House at the Institute of Naval Medicine, Alverstoke. PMID- 10695124 TI - Subcellular imaging by dynamic SIMS ion microscopy. PMID- 10695125 TI - Electrically facilitated molecular transport. Analysis of the relative contributions of diffusion, migration, and electroosmosis to solute transport in an ion-exchange membrane. AB - Electrically facilitated molecular transport in an ion-exchange membrane (Nafion, 1100 equiv wt) has been studied using a scanning electrochemical microscope. The transport rates of ferrocenylmethyltrimethylammonium (a cation), acetaminophen (a neutral molecule), and ascorbate (an anion) through approximately 120-micron thick membranes were measured as a function of the iontophoretic current passed across the membrane (-1.0 to +1.0 A/cm2). Transport rates were analyzed by employing the Nernst-Planck equation, modified to account for electric field driven convective transport. Excellent agreement between experimental and theoretical values of the molecular flux was obtained using a single fitting parameter for each molecule (electroosmotic drag coefficient). The electroosmotic velocity of the neutral molecule, acetaminophen, was shown to be a factor of approximately 500 larger than that of the cation ferrocenylmethyltrimethylammonium, a consequence of the electrostatic interaction of the cation with the negatively charged pore walls of the ion-exchange membrane. Electroosmotic transport of ascorbate occurred at a negligible rate due to repulsion of the anion by the cation-selective membrane. These results suggest that electroosmotic velocities of solute molecules are determined by specific chemical interactions of the permeant and membrane and may be very different from the average solution velocity. The efficiency of electroosmotic transport was also shown to be a function of the membrane thickness, in addition to membrane/solute interactions. PMID- 10695126 TI - Single-molecule identification by spectrally and time-resolved fluorescence detection AB - A method to identify single molecules rapidly and with high efficiency based on simple probability considerations is proposed. In principle, any property of a detected photon in a single-molecule fluorescence experiment, e.g., emission wavelength, arrival time after pulsed excitation, and polarization, can be analyzed within the framework of the outlined methodology. Monte Carlo simulations show that less than 500 photons are needed to assign an observed single molecule to one out of four species with a confidence level higher than 99.9%. We show that single dye molecules of four different dyes embedded in a polymer film can be identified with time-correlated single-photon counting spectrally resolved in two channels. PMID- 10695127 TI - Ion isolation in a continuous zero angle reflecting time-of-flight mass spectrometer AB - A continuous zero angle reflecting time-of-flight mass spectrometer capable of tandem mass spectrometry measurements with high resolution and high sensitivity has been developed. The instrument design features two pulsed-ion mirrors in a coaxial geometry. Ions can be reflected back and forth with the mirrors, which increases the net flight length and permits kinetic energy focusing for enhanced resolution. The instrument also contains an electrostatic particle guide which increases ion transmission efficiency and can be used in a bipolar pulsed mode to isolate ions of interest for structural study. PMID- 10695128 TI - Evaluation of uncompensated solution resistance for electrodes with spherical-cap geometry AB - Typical custom and commercial hanging mercury drop electrodes have the geometry of a spherical cap formed by the plane of the lower surface of the electrode holder cutting off the top of the drop. To conduct accurate quantitative measurements by voltammetry, it is necessary to be able to account for the effects of solution resistance, Ru. A method of determining the solution resistance is proposed and tested. The method involves making measurements of a test reaction (in this case, oxidation of ferrocene) by cyclic voltammetry at scan rates where resistance effects are important and at more than one concentration. When the data are analyzed by digital simulation, it is found that only one value of Ru will provide adequate matches between simulation and experiment at all concentrations. An approximate equation has been derived that allows calculation of Ru from the dimensions of the spherical-cap electrode and the solution resistivity. The calculated values of Ru for electrodes of three different sizes agreed well with the measured values. Error analysis was performed for a particular measurement, the determination of the standard heterogeneous electron-transfer rate constant, ks, by cyclic voltammetry, and it was shown that uncertainty in Ru puts an upper limit of about 1 cm/s for the determination of ks with the hanging mercury drop electrodes used in this study. PMID- 10695129 TI - Absorption of hydrophobic compounds into the poly(dimethylsiloxane) coating of solid-phase microextraction fibers: high partition coefficients and fluorescence microscopy images. AB - The use of solid-phase microextraction with poly(dimethylsiloxane) (PDMS)-coated glass fibers for the extraction and analysis of hydrophobic organic analytes is increasing. The literature on this topic is characterized by large discrepancies in partition coefficients and an uncertainty of whether highly hydrophobic analytes are retained by absorption into the fiber coating or by adsorption to the fiber surface. We applied a new method, which minimizes the impact of experimental artifacts, to determine PDMS water partition coefficients of 17 hydrophobic analytes including chlorinated benzenes, PCBs, PAHs, and p,p'-DDE. These partition coefficients are several orders of magnitude higher than some reported values. Two observations strongly suggest that the retention of hydrophobic organic substances is governed by partitioning into the PDMS coating. (1) The partition coefficients are proportional with octanol/water partition coefficients. (2) The fluorescence of fluoranthene was observed to be homogeneously distributed within the polymer coating when studied by means of fluorescence microscopy. Implications of these findings for the application of solid-phase microextraction with respect to potential detection limits, with respect to biomimetic extraction, and with respect to measurements in multicompartment systems are discussed. PMID- 10695130 TI - Visual and colorimetric lithium ion sensing based on digital color analysis. AB - A new optical analytical method, "Digital Color Analysis (DCA)", is proposed based on a digital color analyzer instead of the conventional optical methodology, "Spectrophotometry". The digital color analyzer is a hand-held-size instrument for measuring "colors", and it can transform the color information into numerical values, color library data, etc., that can be treated as analytical information. DCA gives us a more informative analytical method than spectrophotometry by treating colors as digital information. In addition, DCA can also simulate the optimum color variations for optimization of the visual sensor with computer assistance. By utilizing colors as digital information, colorimetric analysis that has been used for only semiquantitative analysis can serve as an accurate determination method. On the basis of DCA, we developed a plasticized PVC film optode and a paper optode for Li+ determination in saliva. After the optimization of color variation and the detection range for the Li+ measurements, the optode membrane gives colorless gray in the Li+ therapeutic range (at 10(-3) M) in saliva. Consequently, whether or not the optimum therapeutic Li+ concentration is maintained can be easily evaluated with these optodes. Especially, the sensing paper optode can be easily handled within a short measurement time (approximately 80 s) which is suitable for home use. Using the digital color analyzer with QxQy coordinates, a linear relation calibration curve can be obtained over the range from 10(-5) to 10(-1) M Li+, in which the analyzer can detect a concentration difference of approximately 0.1 mM Li+. For the near future, an accurate and simple analysis is needed for a health check at home that does not require going to a hospital. The optode based on DCA has great potential for this analytical purpose. PMID- 10695131 TI - Characterization of pH in liquid mixtures of methanol/H2O/CO2. AB - The presence of H2O and CO2 in enhanced-fluidity liquids changes the pH in these mixtures due to the formation of carbonic acid. The acid-base equilibria in enhanced-fluidity liquids will also be affected by the reduction in the dielectric constant with the addition of CO2. The pH of enhanced-fluidity liquid mixtures at room temperature was determined from the UV/visible absorption spectra of several pH indicators. pH values of methanol/H2O/CO2 mixtures with CO2 proportions as high as 19.2 mol% are reported. The effect of adding buffer to methanol/H2O/CO2 mixtures on pH was also studied. It was also shown that pressure variation did not significantly influence the pH of enhanced-fluidity liquids. PMID- 10695132 TI - Static subcritical water extraction combined with anion exchange disk sorption for determining chlorinated acid herbicides in soil. AB - Static subcritical water extraction (SbWE) was coupled with collection on a strong anion exchange (SAX) disk for the determination of chlorinated acid herbicides and their esters in soil. With 100-150 degrees C water, esters were hydrolyzed into their acid form, and the herbicide acids extracted by subcritical water were trapped onto/into a SAX disk as the extraction cell was cooled. The trapped solutes were then derivatized for gas chromatographic (GC) analysis by placing the disk into a GC autosampler vial containing 1 mL of N,O bis(trimethylsilyl)trifluoroacetamide derivatizing reagent. With the static SbWE/SAX disk extraction, nearly quantitative recoveries (typically over 80%) were obtained at 100 degrees C for 30 min in the extraction of herbicide acids and esters spiked on several different soils covering a range of organic content from 0.3 to 12%. Good agreements were reached between this method and EPA method 8151 for aged spiked soils. Detection limits of the static SbWE/SAX disk extraction were from 0.05 to 0.5 ppm and from 0.01 to 0.5 ppm using GC/electron capture detector and GC/mass spectrometry, respectively. The method is fast and simple and uses a small amount of organic solvent. PMID- 10695133 TI - Real-time amperometric measurements of zeptomole quantities of dopamine released from neurons. AB - Amperometry with carbon-fiber microelectrodes provides a unique way to measure very small chemical concentration changes at the surface of biological cells. In this work, an investigation of dopamine release from individual neurons isolated from the mouse retina is described. The mice were genetically modified so that, in cells that expressed the protein responsible for catecholamine synthesis, tyrosine hydroxylase, the marker protein, placental alkaline phosphatase, was also expressed. This modification allowed for identification of the dopamine containing cells among the many present in the freshly dissociated retina. Release of dopamine was evoked by chemical secretagogues delivered from micropipets that were calibrated with respect to response time and concentration delivered. Amperometric measurements were recorded with low-noise patch clamp amplifiers, and the primary noise source was found to be the electrode capacitance. Dopamine release occurred in the form of transient concentration spikes, consistent with release from small intracellular vesicles. With optimized filtering of the data, the quantity secreted during each release event could be determined. The average quantity determined at one cell was 52 zmol. However, the spikes were quite variable in size and the amount released per event ranged from 8 to 170 zmol. These measurements allow an estimation of the concentration of released transmitter in a synapse. PMID- 10695134 TI - Electrochemistry in nanovials fabricated by combining screen printing and laser micromachining AB - The coupling of screen-printing and laser micromachining technology has been used to create a nanovial with "built-in" working and reference electrodes. The volume of the nanovial was calculated to be 7.2 nL using dimensions determined by SEM. The electrochemical nanovial was characterized using the ferri/ferrocyanide redox couple. Cyclic voltammetry and chronoamperometry experiments were performed with electrochemical nanovials utilizing 5% (v/v) glycerin in the solutions and a humidified headspace to control evaporation of the small-volume samples. Chronoamperometry experiments gave results consistent with a diffusion-limited process and revealed a working electrode surface area of 2.6 x 10(4) micron 2. The ultrasmall-volume cells represent a simple, reliable, low-cost approach for the fabrication of complete electrochemical nanovials. PMID- 10695135 TI - Covalent attachment of osmium complexes to glucose oxidase and the application of the resulting modified enzyme in an enzyme switch responsive to glucose. AB - Pyridine-based osmium complexes bearing either a carboxylate or aldehyde group were covalently attached to glucose oxidase and were shown to work as mediators for the reoxidation of the enzyme. For the complex containing the carboxylate group, the binding was made through carbodiimide coupling to the amine residues in the protein. For the complex containing the aldehyde group, the reductive coupling was carried out by condensation with the amino groups on the protein in the presence of sodium cyanoborohydride. Electrochemical studies show evidence for both intramolecular and intermolecular redox mediation for the electrochemical reoxidation of the modified glucose oxidases in the presence of glucose. The modified enzymes adsorbed on glassy carbon and platinum show different electrochemical responses for the two electrode materials, suggesting that orientation of the adsorbed enzyme is induced due to the interaction of the osmium complex with the different surfaces. Construction of enzyme switches based on these modified enzymes was carried out, and their responses were compared with those obtained using native glucose oxidase and a soluble redox mediator. PMID- 10695136 TI - Dual-pipet techniques for probing ionic reactions AB - Novel dual-pipet electrodes prepared by pulling borosilicate theta-tubing are described. Three types of electrochemical experiments employing such devices include the following: (1) generation/collection experiments in which ions are ejected from one of two micropipets ("generator") into the external solution and collected at the second pipet ("collector"), (2) measurements of ohmic current voltage curves, and (3) ion-transfer voltammetry "in the air". The first setup is used for probing ion transfers at the interface between two immiscible liquids and homogeneous reactions in solution involving ionic species. Such experiments are reported for two model processes, i.e., simple and facilitated transfers of potassium between aqueous and organic phases and complexation of potassium with dibenzo-18-crown-6 in organic solution. The second arrangement is used for characterization of theta-pipets. The last arrangement can be useful for preparation of gas sensors. The possibility of measuring the concentration of volatile substances (e.g., ammonia and nitric acid) in the gaseous phase has been demonstrated. PMID- 10695137 TI - Electrocatalytic detection of NADH and glycerol by NAD(+)-modified carbon electrodes. AB - The electrochemical oxidation of the adenine moiety in NAD+ and other adenine nucleotides at carbon paste electrodes gives rise to redox-active products which strongly adsorb on the electrode surface. Carbon paste electrodes modified with the oxidation products of NAD+ show excellent electrocatalytic activity toward NADH oxidation, reducing its overpotential by about 400 mV. The rate constant for the catalytic oxidation of NADH, determined by rotating disk electrode measurements and extrapolation to zero concentration of NADH, was found to be 2.5 x 10(5) M-1 s-1. The catalytic oxidation current allows the amperometric detection of NADH at an applied potential of +50 mV (Ag/AgCl) with a detection limit of 4.0 x 10(-7) M and linear response up to 1.0 x 10(-5) M NADH. These modified electrodes can be used as amperometric transducers in the design of biosensors based on coupled dehydrogenase enzymes and, in fact, we have designed an amperometric biosensor for glycerol based on the glycerol dehydrogenase (GlDH) system. The enzyme GlDH and its cofactor NAD+ were co-immobilized in a carbon paste electrode using an electropolymerized layer of nonconducting poly(o phenylenediamine) (PPD). After partial oxidation of the immobilized NAD+, the modified electrode allows the amperometric detection of the NADH enzymatically obtained at applied potential above 0 V (Ag/AgCl). The resulting biosensor shows a fast and linear response to glycerol within the concentration range of 1.0 x 10(-6)-1.0 x 10(-4) M with a detection limit of 4.3 x 10(-7) M. The amperometric response remains stable for at least 3 days. The biosensor was applied to the determination of glycerol in a plant-extract syrup, with results in good agreement with those for the standard spectrophotometric method. PMID- 10695138 TI - Atmospheric NO2: in situ laser-induced fluorescence detection at parts per trillion mixing ratios AB - We describe a time-gated laser-induced fluorescence instrument designed for accurate (+/- 5%, 1 sigma), continuous, autonomous, in situ observations of NO2 with the sensitivity (15 ppt/10 s at S/N = 2) and portability necessary to study NO2 anywhere in the troposphere. The technique is advantageous because it is spectroscopically specific and direct in that it does not require conversion of NO2 into another species (e.g., NO) prior to detection, eliminating a class of potential interferences. Performance of the instrument is illustrated with 15 weeks (July-Oct 1998) of observations at the University of California, Blodgett Forest field station located in the foothills of the Sierra Nevada and 4 weeks (June 15-July 15, 1999) in Nashville, TN during the Southern Oxidants Study. Ambient concentrations of NO2 at Blodgett Forest varied from below 50 ppt to 4000 ppt and NO2 ranged from 5 to 50% of the total reactive nitrogen; while in Nashville, TN, concentrations ranged from 1 to 75 ppb. PMID- 10695139 TI - High-resolution mass spectrometry with a quadrupole operated in the fourth stability region AB - The properties of a quadrupole mass filter operated in the fourth stability region with Mathieu parameters (a,q) = (2 x 10(-3), 21.3) have been investigated experimentally and theoretically. A resolution at half-height (R1/2) of 13,900 has been obtained for 40 eV 39K+ ions formed by thermal ionization. With ion energies of 750 eV or more, a resolution at half-height of up to 5000 can be obtained. Extrapolation of resolution data obtained with ions of 40 eV-4800 eV energy shows that unit resolution should be possible for ions with energies of approximately 10 keV. For low-energy ions (e.g., 75 eV), the transmission of the quadrupole operated in the fourth region is many orders of magnitude less than that of a quadrupole operated in the first or second stability regions. Acceptance calculations show this is a result of the combination of a much lower acceptance and strongly defocusing fringing fields. However, for higher ion energies, the transmission with operation in the fourth region is found to be comparable to that of a quadrupole operated at the same resolution in the second region. The implications for inductively coupled plasma mass spectrometry are briefly discussed. PMID- 10695140 TI - Determination of chlorophenols in soils using accelerated solvent extraction combined with solid-phase microextraction. AB - A method for the determination of chlorophenols in soil samples using accelerated solvent extraction (ASE) with water as the solvent combined with solid-phase microextraction (SPME) and GC/MS has been developed. Important ASE parameters, such as extraction temperature and time, were optimized using a spiked wetland soil. The effect of small amounts of organic modifiers on the extraction yields was studied. An extraction temperature of 125 degrees C and 10 min extractions performed three times proved optimal. Two ASE-SPME procedures without and with an organic modifier (5% acetonitrile) were evaluated with respect to precision and detection limits (LOD). The reproducibility of replicate water extractions/SPME determinations (n = 6) was in the range 7-20% relative standard deviation for the nine chlorophenols investigated. LOD values in the low-ppb range were achieved for all chlorophenols. The ASE-SPME procedure presented here was applied to the determination of chlorophenols in soil samples taken from contaminated areas near Bitterfeld, Germany. PMID- 10695141 TI - The characteristics of peptide collision-induced dissociation using a high performance MALDI-TOF/TOF tandem mass spectrometer. AB - A new matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time-of flight (TOF/TOF) high-resolution tandem mass spectrometer is described for sequencing peptides. This instrument combines the advantages of high sensitivity for peptide analysis associated with MALDI and comprehensive fragmentation information provided by high-energy collision-induced dissociation (CID). Unlike the postsource decay technique that is widely used with MALDI-TOF instruments and typically combines as many as 10 separate spectra of different mass regions, this instrument allows complete fragment ion spectra to be obtained in a single acquisition at a fixed reflectron voltage. To achieve optimum resolution and focusing over the whole mass range, it may be desirable to acquire and combine three separate sections. Different combinations of MALDI matrix and collision gas determine the amount of internal energy deposited by the MALDI process and the CID process, which provide control over the extent and nature of the fragment ions observed. Examples of peptide sequencing are presented that identify sequence-dependent features and demonstrate the value of modifying the ionization and collision conditions to optimize the spectral information. PMID- 10695142 TI - Field-emission cold-cathode el source for a microscale ion trap mass spectrometer AB - A cold-cathode electron impact ionization source based on field emission from an array of diamond-coated silicon whiskers is described. The source is coupled to a microscale ion trap mass spectrometer (r0 = 0.50 mm, z0 = 0.50 mm). An electron beam of 250 nA could be obtained through the 0.45-mm diameter opening in the end cap electrode. PMID- 10695143 TI - Electron capture dissociation for structural characterization of multiply charged protein cations. AB - For proteins of < 20 kDa, this new radical site dissociation method cleaves different and many more backbone bonds than the conventional MS/MS methods (e.g., collisionally activated dissociation, CAD) that add energy directly to the even electron ions. A minimum kinetic energy difference between the electron and ion maximizes capture; a 1 eV difference reduces capture by 10(3). Thus, in an FTMS ion cell with added electron trapping electrodes, capture appears to be achieved best at the boundary between the potential wells that trap the electrons and ions, now providing 80 +/- 15% precursor ion conversion efficiency. Capture cross section is dependent on the ionic charge squared (z2), minimizing the secondary dissociation of lower charge fragment ions. Electron capture is postulated to occur initially at a protonated site to release an energetic (approximately 6 eV) H. atom that is captured at a high-affinity site such as -S-S- or backbone amide to cause nonergodic (before energy randomization) dissociation. Cleavages between every pair of amino acids in mellitin (2.8 kDa) and ubiquitin (8.6 kDa) are represented in their ECD and CAD spectra, providing complete data for their de novo sequencing. Because posttranslational modifications such as carboxylation, glycosylation, and sulfation are less easily lost in ECD than in CAD, ECD assignments of their sequence positions are far more specific. PMID- 10695144 TI - Mass spectrometric analysis of mercury incorporation into proteins for X-ray diffraction phase determination. AB - Heavy-atom incorporation is an essential and often rate-limiting step in the determination of phases for X-ray diffraction studies of protein structures. Until the present, there has been no practical method (short of the X-ray diffraction experiment itself) to judge the success and extent of incorporation. Here we show that mass spectrometry is an effective tool for determining the extent of heavy-atom incorporation in proteins. In particular, we demonstrate the utility of matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and electrospray ionization mass spectrometry (ESI-MS) for assaying mercury derivatization of cysteinyl thiol groups in proteins. Each of these mass spectrometric methods has advantages and drawbacks. ESI-MS provides a more accurate quantitative measurement of the extent of mercury incorporation, while MALDI-MS provides a useful lower limit to the level of mercury incorporation. Conversely, MALDI-MS does not require removal of excess derivatization reagents, salts and buffers, thus permitting facile analysis of single protein crystals as well as rapid, semiquantitative evaluation of the extent of protein mercuration. The approaches described in the present paper have contributed to the successful X-ray analyses of several noteworthy protein structures. PMID- 10695145 TI - Using different drift gases to change separation factors (alpha) in ion mobility spectrometry AB - The use of different drift gases to alter separation factors (alpha) in ion mobility spectrometry has been demonstrated. The mobility of a series of low molecular weight compounds and three small peptides was determined in four different drift gases. The drift gases chosen were helium, argon, nitrogen, and carbon dioxide. These drift gases provide a range of polarizabilities and molecular weights. In all instances, the compounds showed the greatest mobility in helium and the lowest mobility in carbon dioxide; however the percentage change of mobility for each compound was different, effectively changing the alpha value. The alpha value changes were primarily due to differences in drift gas polarizability but were also influenced by the mass of the drift gas. In addition, gas-phase ion radii were calculated in each of the different drift gases. These radii were then plotted against drift gas polarizability producing linear plots with r2 values greater than 0.99. The intercept of these plots provides the gas-phase radius of an ion in a nonpolarizing environment, whereas the slope is indicative of the magnitude of the ion's mobility change related to polarizability. It therefore, should be possible to separate any two compounds that have different slopes with the appropriate drift gas. PMID- 10695146 TI - Trapping of bead-based reagents within microfluidic systems: on-chip solid-phase extraction and electrochromatography AB - A 330-pL chromatographic bed was fabricated on a glass substrate as part of an electroosmotically pumped microfluidic system. Two weirs within a sample channel formed a cavity in which octadecylsilane (ODS) coated silica beads (1.5-4 microns diameter) were trapped. ODS beads were mobilized into and out of the cavity using electroosmotic flow through a bead-introduction channel which accessed the cavity. This procedure allowed the beads in the cavity to be repeatedly exchanged. A 1 nM solution of a nonpolar analyte (BODIPY 493/503) in buffer was loaded onto the beads for different lengths of time using an electroosmotic flow of 1.2 nL/s. The material retained on the ODS phase was then eluted by electroosmotic flow of acetonitrile with a concentration enhancement of up to 500 times. Capillary electrochromatography was conducted using a similar device. BODIPY and fluorescein were loaded onto a 200-micron-long chromatographic bed and then separated in an isocratic CEC run with an acetonitrile/buffer mobile phase. Complete separation was achieved in less than 20 s with a 2-micron plate height. PMID- 10695147 TI - Peak capacity, peak-capacity production rate, and boiling point resolution for temperature-programmed GC with very high programming rates AB - Recent advances in column heating technology have made possible very fast linear temperature programming for high-speed gas chromatography. A fused-silica capillary column is contained in a tubular metal jacket, which is resistively heated by a precision power supply. With very rapid column heating, the rate of peak-capacity production is significantly enhanced, but the total peak capacity and the boiling-point resolution (minimum boiling-point difference required for the separation of two nonpolar compounds on a nonpolar column) are reduced relative to more conventional heating rates used with convection-oven instruments. As temperature-programming rates increase, elution temperatures also increase with the result that retention may become insignificant prior to elution. This results in inefficient utilization of the down-stream end of the column and causes a loss in the rate of peak-capacity production. The rate of peak-capacity production is increased by the use of shorter columns and higher carrier gas velocities. With high programming rates (100-600 degrees C/min), column lengths of 6-12 m and average linear carrier gas velocities in the 100-150 cm/s range are satisfactory. In this study, the rate of peak-capacity production, the total peak capacity, and the boiling point resolution are determined for C10 C28 n-alkanes using 6-18 m long columns, 50-200 cm/s average carrier gas velocities, and 60-600 degrees C/min programming rates. It was found that with a 6-meter-long, 0.25-mm i.d. column programmed at a rate of 600 degrees C/min, a maximum peak-capacity production rate of 6.1 peaks/s was obtained. A total peak capacity of about 75 peaks was produced in a 37-s long separation spanning a boiling-point range from n-C10 (174 degrees C) to n-C28 (432 degrees C). PMID- 10695148 TI - Separation and identification of peptides from gel-isolated membrane proteins using a microfabricated device for combined capillary electrophoresis/nanoelectrospray mass spectrometry. AB - The coupling of microfabricated devices to nanoelectrospray mass spectrometers using both a triple quadrupole and a quadrupole time-of-flight mass spectrometer (QqTOF MS) is presented for the analysis of trace-level membrane proteins. Short disposable nanoelectrospray emitters were directly coupled to the chip device via a low dead volume connection. The analytical performance of this integrated device in terms of sensitivity and reproducibility was evaluated for standard peptide mixtures. A concentration detection limit ranging from 3.2 to 43.5 nM for different peptides was achieved in selected ion monitoring, thus representing a 10-fold improvement in sensitivity compared to that of microelectrospray using the same chip/mass spectrometer. Replicate injections indicated that reproducibility of migration time was typically less than 3.1% RSD whereas RSD values of 6-13% were observed on peak areas. Although complete resolution of individual components is not typically achieved for complex digests, the present chip capillary electrophoresis (chip-CE) device enabled proper sample cleanup and partial separation of multicomponent samples prior to mass spectral identification. Analyses of protein digests were typically achieved in less than 1.5 min with peak widths of 1.8-2.5 s (half-height definition) as indicated from individual reconstructed ion electropherograms. The application of this chip CE/QqTOF MS system is further demonstrated for the identification of membrane proteins which form a subset of the Haemophilus influenzae proteome. Bands first separated by 1D-gel electrophoresis were excised and digested, and extracted tryptic peptides were loaded on the chip without any further sample cleanup or on line adsorption preconcentration. Accurate molecular mass determination (< 5 ppm) in peptide-mapping experiments was obtained by introducing an internal standard via a postseparation channel. The analytical potential of this integrated device for the identification of trace-level proteins from different strains of H. influenzae is demonstrated using both peptide mass-fingerprint database searching and on-line tandem mass spectrometry. PMID- 10695149 TI - Silicate polymerization for the preparation of bed-retention frits in capillary electrochromatography AB - Bubble formation, which is associated with bed-retention frits, is a critical experimental problem in capillary electrochromatography systems. In this investigation, porous silica frits were prepared via spot-heating of a silicate solution, and the effects of several experimental parameters on their performance were studied. The optimal sodium silicate concentrations were 10.8% and 5.4% (w/v) for outlet and inlet frits, respectively. The heating times were 5-6 s for outlet frits and < 1 s for inlet frits. Under optimized conditions, outlet frits were 75 microns (+/- 12 microns) and the heat treatment did not make the capillary fragile at the frit location. Bubble formation was affected by frit length, density, and silanization of the frits with trimethylchlorosilane. Packed capillaries with optimized frits were used successfully in a commercial CE instrument over a normal working day without pressurization, at relatively high ionic strengths (10 mM), and over a wide range of acetonitrile compositions (20% 80%). Currents were also stable for > or = 3 h under very high current (27 microA) conditions. As part of this study, the efficiency and reproducibility of packed capillaries were also briefly evaluated. PMID- 10695150 TI - Capillary electrochromatography using a strong cation-exchange column with a dynamically modified cationic surfactant AB - A novel mode of capillary electrochromatography (CEC), called dynamically modified strong cation-exchange CEC (DMSCX-CEC), is described in this paper. A column packed with a strong cation-exchange (SCX) packing material was dynamically modified with a long-chain quaternary ammonium salt, cetyltrimethylammonium bromide (CTAB), which was added to the mobile phase. CTAB ions were adsorbed onto the surface of the SCX packing material, and the resulting hydrophobic layer on this packing was used as the stationary phase. Using the dynamically modified SCX column, neutral solutes were separated with the CEC mode. The highest number of theoretical plates obtained was about 190,000/m, and the relative standard deviations (RSD's) for migration times and capacity factors of alkylbenzenes were less than 1.0% and 2.0% for five consecutive runs, respectively. The effects of CTAB and methanol concentrations and the pH value of the mobile phase on the electroosmotic flow and the separation mechanism were investigated. Excellent simultaneous separation of the basic and neutral solutes in DMSCX-CEC with a high-pH mobile phase was obtained. A mixture containing the acidic, basic, and neutral compounds was well separated in this mode with a low-pH mobile phase; however, peak tailing for basic compounds was observed in this mobile phase. PMID- 10695151 TI - Measurement of aqueous Henry's law constants for oxygenates and aromatics found in gasolines by the static headspace method AB - A simple equilibrium vessel made from a large stopcock was used in conjunction with the static headspace method to measure the aqueous Henry's law constants for methyl tert-butyl ether (MTBE) and other potential oxygenates as well as certain of the aromatics found in gasoline. The present method involves using only one aqueous solution whose concentration is not necessarily known. The only quantity that must be known is the phase ratio. Plotting a series of peak areas of the analyte in the headspace after each equilibrium change against the initial peak area of the analyte found in the headspace leads to a linear relationship whose slope is dependent on the analyte's unitless Henry's law constant and the phase ratio. This new method is found to yield unitless aqueous Henry's law constants in the range from 0.02 to 0.2, which are especially applicable to the oxygenates and aromatics found in gasolines. PMID- 10695152 TI - Sheathless capillary electrophoresis/electrospray mass spectrometry using a carbon-coated fused-silica capillary AB - A simple procedure was developed for preparing a carbon-coated fused-silica capillary for use in sheathless capillary electrophoresis/electrospray mass spectrometry (CE/ESI-MS). The tapered capillary tip was smeared with a marker pen before coating with carbon using a soft pencil. The layer from the ink of the marker pen was critical to the preparation of the carbon-coated capillary. The fabrication of a carbon-coated fused-silica capillary tip requires less than 1 min. The stability of this carbon-coated fused-silica capillary is examined, and its utility in on-line sheathless CE/ESI-MS is demonstrated with the separation of berberine, coptisine, and palmatine chlorides. Although the carbon-coated fused-silica capillary tip is not as rugged as a gold-coated capillary, it is durable enough for sheathless CE/ESI-MS applications. Moreover, it is easy to refurbish the column once the performance of the tip is degraded. PMID- 10695153 TI - In-line catalytic purification of carbon dioxide used in analytical-scale supercritical fluid extraction AB - Supercritical fluid extraction analyses are often compromised by trace impurities present in the solvent carbon dioxide. These impurities, commonly used as lubricants in the specialty gas industry, can produce significant background levels, increasing limits of detection and quantification. This problem is especially severe when electron capture detection (ECD) is used for trace concentrations of analytes (e.g., polychlorinated biphenyls and chlorinated pesticides). In this study, an in-line catalyst-based purification system was successfully employed to remove ECD-responsive contaminants from CO2. Low-purity (98%) "Bone Dry" CO2 was purified to levels cleaner than a very-high-purity grade of CO2 specified at less than 10 ppt ECD-responsive contaminants. Purification was successfully applied to extremely sensitive on-column experiments as well as higher flow rate off-line experiments. In addition to lowering limits of detection and quantification, significant cost savings can be realized by purifying inexpensive, low-purity CO2 instead of relying on much more expensive, prepurified CO2. PMID- 10695154 TI - Determination of a sulfur-containing drug in human plasma by an improved method for sulfur chemiluminescence detection in combination with capillary gas chromatography. AB - An improved method for sulfur chemiluminescence detection in combination with capillary gas chromatography was established. The method was applied to the analysis of a sulfur-containing antiasthma drug, S-1452, and its nine metabolities in human plasma. The high selectivity and sensitivity of the method allowed accurate quantitation of trace levels of these compounds in human plasma with no interferences from the biological components present. To enable stable measurements and maintain reproducibility over a long period, the performance characteristics of a commercially available instrument were investigated. The gas seal in the injection port was found to easily corrode through interaction of the sulfur analyte with the metal gas seal. To prevent this, a disk made from an alloy of platinum and gold (60:40) was mounted on the gas seal. The reproducibility of the measurement was improved remarkably by using the disk. The use of compressed air of high purity significantly lowered the signal-to-noise ratio. The optical filter was kept clean by using a nickel catalyst to trap ozone in place of copper manganese oxide (CuMn2O4). These improvements raised the sensitivity and selectivity with the lower quantitation limits of 0.5-1.0 ng/mL in human plasma. PMID- 10695155 TI - Comments on "adsorption versus absorption of polychlorinated biphenyls onto solid phase microextraction coatings" PMID- 10695156 TI - Women and bipolar disorder: an update. AB - The major clinical challenges facing women with bipolar illness, and the practitioners who care for them, are the management of rapid cycling and of the postpartum period. Among patients with rapid cycling bipolar disorder, the treatment of depression is particularly problematic. The most commonly prescribed mood stabilizers are more potent antimanic than antidepressant agents, and the use of antidepressants may precipitate mania or rapid cycling. The availability of new anticonvulsant medications that may have both mood-stabilizing and antidepressant effects is an important development in this regard. In the postpartum period, women with bipolar illness are at uniquely high risk for relapse. The possible reasons for this high risk, and options for the management of pregnancy and the postpartum period in bipolar women, are discussed. PMID- 10695157 TI - Body dysmorphic disorder: diagnostic controversies and treatment challenges. AB - Body dysmorphic disorder (BDD) is a distressing, impairing, and relatively common yet underrecognized disorder. This clinically focused article discusses the following diagnostic controversies and challenges associated with BDD: the underdiagnosis and misdiagnosis of BDD, the relationship between BDD and obsessive-compulsive disorder, the relationship between BDD and depression, the delusionality controversy, and whether BDD can be diagnosed in children and adolescents. This article also discusses treatment controversies and challenges associated with serotonin reuptake inhibitors, antipsychotics, cognitive behavioral therapy, approaches to treatment-resistant BDD, and getting reluctant patients to accept psychiatric treatment. PMID- 10695158 TI - Workplace violence and psychiatric practice. AB - The authors provide an overview of what is known about workplace violence and discuss how to deal with issues of workplace violence that arise in clinical practice. They review myths and facts about workplace violence, including research on prevention. Legal issues relating to the psychiatrist as employer and the Americans with Disability Act are presented. General principles of violence assessment are reviewed and the authors then discuss the psychiatrist as consultant to the workplace and as clinician treating a victim or perpetrator of workplace violence. Three cases illustrate the general principles provided. PMID- 10695159 TI - Avoiding psychotropic drug interactions in the cardiovascular patient. AB - Recent advances in our understanding of the hepatic cytochrome P450 inhibitory effects of the newer antidepressants have increased concern about drug interactions in the practice of psychiatry. The authors summarize the potential interactions of psychoactive drugs with cardiovascular medications. Practicing psychiatrists encounter many patients with cardiovascular disease, and an awareness of these interactions will improve knowledgeable prescribing for medically ill patients with comorbid mental disorders. PMID- 10695160 TI - New antipsychotic medications: more than old wine and new bottles. AB - Four new antipsychotic medications--clozapine, risperidone, olanzapine, and quetiapine--have been introduced in the United States during the past decade. These new medications now account for the majority of antipsychotic prescriptions. The author reviews specific issues related to the use of traditional antipsychotic medications and then highlights the emerging clinical research data regarding the new medications, which have all been shown to be efficacious in the treatment of schizophrenia. Clinical research data indicate that they are also more useful for a broader array of symptoms associated with schizophrenia than traditional compounds. Furthermore, movement disorder side effects are substantially decreased--a property that leads to higher acceptability. Surprisingly, there has been little relationship between the pivotal trials designed for FDA approval and current dosing strategies in broader clinical settings. These dosing issues are described. New uses, including treatment of mood disorders and conduct disorder, are also discussed. These medicines offer substantial hope for improved treatment of schizophrenia. PMID- 10695161 TI - Priming and projective identification. AB - The sense of coercion that is a central feature of projective identification might be understood in terms of the recently discovered phenomenon of priming. Studies of priming show that a subject may record and be influenced by stimuli of which he or she is not conscious. Becoming aware of such influence is of central therapeutic significance. Certain priming experiments suggest that Freud's discipline of "evenly suspended attention" may provide a means of potentiating this awareness. PMID- 10695162 TI - Autonomy and relatedness in the development of anorexia nervosa: a clinical case series using grounded theory. AB - The authors propose a prototype for the development of anorexia nervosa on the basis of a series of clinical interviews with 11 women with this disorder. Grounded Theory analysis of the narrative data-generated themes suggested the following prototype: Women with the personality characteristics of compliance and perfectionism who are encouraged by families and culture to substitute others' needs for their own are at greater risk for anorexia nervosa during periods of developmental stress. This proposal is considered from the perspective of psychodynamic, cognitive, and family systems viewpoints as well as attachment theory and feminist models of the development of women. The authors suggest that narrative research contributes substantially to clinical understanding by capturing the intricate details of a phenomenon or the richness of a lived life. Nevertheless, there is a need for further quantitative research to test the specificity and generalizability of this prototype. PMID- 10695163 TI - Magnum miraculum est homo. PMID- 10695164 TI - Client perspectives on occupational therapy practice: are we truly client centred? AB - In Canada, the guidelines for the practice of occupational therapy are named and framed as client-centred. Two in-depth interviews were conducted with clients of mental health services about their experiences with a hospital-based occupational therapy service. These occupational therapy clients described their experiences as prescriptive, and as less than client-centred. With the publication of Enabling occupation: A Canadian occupational therapy perspective (Canadian Association of Occupational Therapists [CAOT], 1997) and an increasingly refined focus on being client-centred, these interviews highlight the challenges of a client-centred practice within the current health care environment. These occupational therapy clients raise issues of importance for occupational therapy. The participants stated that the prescription of 'activity', a lack of choice, and a focus upon the illness as opposed to the individual, served to diminish any collaborative partnership with the client and eliminate the client from any decision-making process. This distancing from the client, in their opinion, served to greatly diminish any therapeutic value of occupation. The participants recommended a greater focus upon occupational choice, consideration of the individual within the client, providing accepting, supportive environments, and using professional expertise on occupation to guide the client towards participation in meaningful occupation. These recommendations are strikingly similar to the most recent guidelines for the client-centred practice of occupational therapy in Canada. A discussion of the implications of these findings for the client-centred practice of occupational therapy is offered. PMID- 10695165 TI - Barriers to client-centredness and their resolution. AB - This research sought to determine which therapist barriers prevent client-centred practice the most, and which methods are perceived as being most effective in resolving therapist barriers. A list of barriers that therapists bring to client centred practice and methods to resolve these was identified from the literature and formed the basis of a questionnaire sent to 60 occupational therapists in the United Kingdom. The results showed that the therapist and client having different goals was the barrier which most prevented client-centred practice. The high ratings of other statements suggested that the values, beliefs and attitudes of therapists and of the employment culture make client-centred practice uncomfortable to use and hence prevent its implementation. Case examples showing how to practice in a client-centred fashion were rated as the most effective method of barrier removal. PMID- 10695166 TI - Validity and community utility of the Canadian Occupational Performance Measure. AB - This study addressed the validity and community utility of the Canadian Occupational Performance Measure (COPM) (Law et al., 1991; 1994; 1998): a measure that now represents a national standard in clinical practice and research in occupational therapy in Canada. The study employed a crossectional design. Participants for the study were former consumers of occupational therapy services, recruited from the Queen's University catchment area (Kingston, North Bay, Oshawa, Perth, Peterborough). A sample of 61 disabled individuals living in the community were recruited. Each individual was sent a package of self administered measures including the Satisfaction with Performance Scaled Questionnaire, the Reintegration to Normal Living Index, the Life Satisfaction Questionnaire, and the Perceived Problems List. An interview was also arranged with the project coordinator, which was based on the COPM and the Consumer Utility Questionnaire. Multivariate analyses showed that construct validity was supported; scores on the COPM were significantly related to theoretically related constructs: satisfaction with performance, reintegration to normal living and life satisfaction. In addition, criterion validity was supported. A majority of participants (53%), when asked about problems of daily living, spontaneously reported at least one of the problems raised on the COPM. Community utility was evaluated highly by participants, 75% of whom found the COPM useful in identifying and rating their problems, and 100% of whom reported no problems in understanding the COPM. PMID- 10695167 TI - [Utilization of lifting equipment and slings in occupational therapy]. AB - The occupational therapist is frequently involved in the allocation process of lifting devices for clients with severe physical disabilities who are living in the community. The aim of this paper is to introduce a conceptual framework to help therapists prescribe lifting devices, including the slings. First, factors influencing the decision to prescribe such an aid are analysed based on the concepts of the Canadian Model of Occupational Performance (Canadian Association of Occupational Therapists, 1997). When working with clients toward maximizing their transfer skills, occupational therapists will take into account different aspects such as the characteristics of the client, the environment and the equipment, as well as the time allocated to complete the activity. Secondly, the notion of the work situation outlined by an organization specializing in work safety measures is used as a guide for transfer evaluation. From this viewpoint, the introduction of the lifting device occurs along a continuum of progressive loss of independence and is determined by degrees of personal independence, human assistance as well as technical assistance required to perform transfers. Finally, advantages and disadvantages of using lifting devices in a home setting are presented as a conclusion to the study. PMID- 10695168 TI - A conceptual model for the development of professional behaviours in occupational therapists. AB - The ever-changing, dynamic practice environment coupled with increased consumer needs and awareness create an atmosphere that requires optimal professionalism from occupational therapists. Professionalism requires specific knowledge, attitudes, and values--all manifested by professional behaviours. The authors assume that professional behaviours mature through a natural developmental process; a process that requires careful nurturing on the part of educators and clinical supervisors. Based on this assumption, the authors propose this conceptual model based on Erikson's life cycle stages. The model implies that occupational therapy professional behaviours develop sequentially through stages that begin during the educational process of occupational therapists, and progress throughout their career. The purpose of this model is to provide a framework for educators and supervisors to nurture professional behaviours in students and novice clinicians, and to continue their own professional growth. PMID- 10695169 TI - Development of a tool to measure clinical competence in occupational therapy: a pilot study? AB - Clinical competence is generally defined as a combination of knowledge, skill and professional behaviour. It is typically assessed using written tests, direct observation, chart audit, client satisfaction surveys and supervisor ratings. This paper describes the development and evaluation of a chart-stimulated recall (CSR) measure that combines the methods of chart audit and clinician interview to assess the clinical competence of practicing occupational therapists. The CSR tool was developed using the Canadian Guidelines for Client-Centred Practice and taps global domains of competence: use of theory, assessment, program planning, intervention, discharge planning, follow-up, program evaluation, clinical reasoning and professional behaviours. This pilot study involved two independent raters/interviewers who assessed twelve occupational therapy clinicians on two occasions using a random sample of client cases/records on each occasion Results indicate that the CSR tool is not only reliable and valid, but also sufficiently generic to be used in a variety of practice settings as a global measure of on the-job performance. PMID- 10695170 TI - Reflections on a renaissance of occupation. AB - At the close of the 20th century, there is a renaissance of occupation in occupational therapy and occupational science. Kielhofner (1992) offers an intraprofessional explanation that the growing interest in occupation recaptures occupational therapy's lost identity. An extraprofessional explanation is that postmodern ideas and social practices have helped to create a societal context in which a renaissance of occupation is welcome. Postmodernism raises questions and awareness of power, diversity, temporality, and situatedness in which normative ideas of occupation as paid work can be challenged. Since occupation is of primary concern to occupational therapy and occupational science, the authors reflect on postmodernism and its influence on a renaissance of occupation in these two fields. These reflections consider what such a renaissance means for occupational therapists and occupational scientists in the 21st century. PMID- 10695171 TI - The occupation of gardening in life-threatening illness: a qualitative pilot project. AB - This qualitative pilot study was completed as an exploratory study of the meaning of gardening using attention restoration theory. Three women with breast cancer who garden for leisure were recruited from a cancer support group. Each participant was interviewed at her home on two occasions. To complement the qualitative data, participants also completed the Perceived Restorativeness Scale (PRS) (Hartig, Korpela, Evans, & Garling, 1996). The interviews revealed six major themes some of which were concerned with the interactions between the gardener and the garden, and others which focused on gardening within the context of having cancer. The qualitative and quantitative outcomes supported the perspective of attention restoration theory. Spirituality was interwoven throughout the comments of two participants but was less important for one participant. The implications of this study for practice and future research are discussed. PMID- 10695172 TI - Ethical decision-making made easier. The use of decision trees in case management. AB - Case managers have never before faced the multitude of difficult ethical dilemmas that now confront them daily. Legal, medical, social, and ethical considerations often fly in the face of previously reliable intuitions. The importance and urgency of facing these dilemmas head-on has resulted in clear calls for action. What are the appropriate legal, ethical, and professional parameters for effective decision making? Are normatively sensitive, but also practically sensible protocols possible? In an effort to address these concerns, Alternatives for the Older Adult, Inc., Rock Island, Illinois established an ethics committee to look into possible means of resolving or dissolving commonly occurring dilemmas. As a result of year-long deliberations, the committee formulated a decision-making strategy whose central apparatus is the decision tree--a flowchart of reasonable decisions and their consequent implications. In this article, we explore the development of this approach as well as the theory that underlies it. PMID- 10695173 TI - Cost effectiveness of a high-risk pregnancy program. AB - This article presents an evaluation of an innovative community-based, case management program for high-risk pregnant women and their infants. A 7-year analysis of the Medicaid claims from 182,196 pregnant women and those for 140,088 infants was conducted. The findings showed improved birth outcomes and a steady decrease in the cost of care for both pregnant women and their infants. Recommendations are made concerning implementation of this program in other settings with other clients. PMID- 10695174 TI - Off the beaten track: MSN education for a changing health care environment. AB - Given the dynamic nature of the health care environment, what should be the focus of graduate education in nursing? New advanced practice roles for nurses are emerging, but few university-based educational programs exist to respond to the need. In developing a Master's program, Baylor University School of Nursing accepted the challenge to deviate from tradition by preparing a nurse who has the advanced knowledge and skills necessary to proactively address the present and future needs of health care in multiple settings and roles. This article discusses this unique and creative program in patient care management. Outcome evaluation reveals that students are attaining the terminal objectives, meeting the outcome criteria, and readily securing employment. PMID- 10695175 TI - Expedited service delivery: helping case managers to increase access to services for the poorest older adults. AB - Low-income older adults have historically received institutionally based care when they were unable to care for themselves and there was a lack of sufficient formal and informal support to allow them to remain in their own homes. In 1981, revision of federal Medicaid legislation permitted states to provide home- and community-based services (HCBS) for older adults at risk of nursing facility placement in an attempt to prevent unnecessary placement and to offer choice. However, Medicaid-HCBS applicants may have to wait up to 45 days or longer for approval of their financial application. During those 45 days, the applicant may enter a nursing facility because Medicaid-HCBS was not available soon enough to prevent placement. This article presents research on an instrument to help case managers initiate community-based in-home services for the lowest-income Medicaid applicants within 3 to 5 days of initial assessment. PMID- 10695176 TI - The Medicare home health benefit implications of recent payment changes. AB - The interim payment system (IPS) for Medicare home health services, enacted in the Balanced Budget Act of 1997, was intended to slow the growth of home health expenses until HCFA could design a new prospective system. Instead, the IPS has acted like a per-case payment system without case-mix adjustment. Its impact on agencies, along with other policy pressures, has been first to slow and then to reverse the dramatic expansion of the home health sector. In this paper, we identify the impetus for payment changes in the recent history of the Medicare home health benefit. We then present emerging evidence about the effects of IPS and other recent policies on home health. Finally, we draw several lessons from this experience for the impending prospective payment system. PMID- 10695177 TI - Determining the relevance of a certification exam to home health care nursing practice. AB - Home health care is enjoying increased use and popularity. Unfortunately, in today's cost-cutting environment, home healthcare is also subject to increased scrutiny and inevitable reimbursement limitations. This is borne out by the impact on home healthcare as a result of the Balanced Budget Act of 1997. Berke (1998) reports that those at greatest risk for cutbacks in care are those that can least afford it--the oldest, sickest, poorest, and most frail. Compounding the financial dilemma that home health care clients face are multiple providers of care, an unrealistic media presentation of health care, and less time for anyone to provide psychosocial-focused care (Simmons, 1990). Home health care clients have a desperate need for an advocate to provide expert navigation through the health care system. Home health care providers are aware of and often responsible for bridging gaps in health, medical benefits, and social services. This article describes a process for determining the relevance of a certification to community nursing clinical practice--using the Advanced Certification in Continuity of Care (A-CCC) exam as the example. PMID- 10695178 TI - Designing through their eyes. Group homes for persons with Alzheimer's disease. PMID- 10695179 TI - Preserving a healthy home. PMID- 10695180 TI - Home modifications. Enabling environments, LLC, a fee-based service. PMID- 10695181 TI - Project Metropair. A free home safety and security program in New York City. PMID- 10695182 TI - Iron and alcohol content of traditional beers in rural Zimbabwe. AB - OBJECTIVES: To determine the concentrations of iron and alcohol in traditional beer, as well as how these may be related to the brewing process. DESIGN: Cross sectional study. SETTING/SUBJECTS: Rural communities living in four of Zimbabwe's nine provinces. MAIN OUTCOME MEASURES: Ionic iron concentration and alcohol concentration in 94 different types of alcoholic beverages prepared in rural areas, and 18 commercially produced beers. RESULTS: The commonest types of traditional beer were a seven day beverage called 'doro rematanda', a by-product of this seven day beer called 'muchaiwa,' and a one-day beverage called 'chikokiyana'. Methods of preparation were similar in the four provinces. Median (Q1, Q3) ionic iron concentrations were 52 (31 to 75) mg/L for the seven-day beer (n = 51), 24 (18 to 36) mg/L for muchaiwa (n = 30) and 21 (17 to 63) mg/L for chikokiyana (n = 13). In contrast, ionic iron concentrations in 12 samples of commercially prepared clear beers were 0.1 mg/L and in commercial opaque beer were 3.6 mg/L. Mean (SD) alcohol concentration in traditional beer was 4.1 g/100 ml (+/- 0.873) compared to 2.8 g/100 ml +/- 1.394) in the muchaiwa and 3.6 g/100 ml (+/- 1.445) in the one day brew, chikokiyana. Mean alcohol concentrations in the three commercial beers are reportedly 3.5 g/100 ml in the opaque beer (Scud), and 4.7 to 5.0 g/ml in clear beer (Zambezi and Castle lagers). CONCLUSIONS: Several preparation methods lead to traditional fermented beverages with very high iron concentrations. Measures to prevent dietary iron overload should include all of these beverages in their scope. PMID- 10695183 TI - Bioavailability of rifampicin in a separate formulation and fixed dose combination with isoniazid NIH: a case for a fixed dose combination (FDC) for the treatment of tuberculosis. AB - OBJECTIVES: To study and compare the bioavailability of rifampicin (RIF), in two locally manufactured formulations; an FDC and a separate formulation and an imported FDC formulation. DESIGN: Open within subjects, single blind cross over study. INTERVENTIONS: Each volunteer subject, acting as their own control, received the two fixed dose combinations and the separate formulation with the same amount of 450 mg RIF. MAIN OUTCOME MEASURES: Cmax (peak drug concentration achieved), Tmax (time at which peak drug concentration is achieved), T1/2el (biological half-life of elimination) and area under the curve (AUC) for zero to 10 hours and zero to infinity. These are obtained from plotting plasma concentration against time. RESULTS: There was a significant difference in the Cmax between free and RIF combined with INH (6.1 and 7.6 mg/l respectively) and no significant difference in the other parameters measured, of the local products. Comparison of the local products and imported product showed no significant difference in AUC but significant differences in T1/2el, C max and Tmax (p = 0.003, 0.041 and 0.025 respectively). CONCLUSION: The Zimbabwe manufactured and the German products had "demonstrable bioavailability" as defined by the International Union Against Tuberculosis and Lung Diseases (IUATLD). The local manufacturer appeared to have the technological capability to produce a registrable combined RIF/INH table to be used in the treatment of tuberculosis and to prevent the irrational use of RIF. PMID- 10695184 TI - Aero-allergen sensitisation patterns amongst atopic Zimbabwean children. AB - OBJECTIVE: To characterize children presenting with atopic conditions using the RAST test. DESIGN: Retrospective descriptive study. SETTING: General paediatric clinic in the private sector. SUBJECTS: 84 children aged below 12 years, who had the RAST test, who presented to a general paediatric clinic between 1993 and 1998 with atopic conditions for care. RESULTS: The median age for all children in the study was 52 months. Forty eight were male and 36 female. Eczema (33.9%) was the most frequent clinical diagnosis especially in those less than 24 months of age, followed by asthma (25.5%), allergic conjunctivitis (24.0%) and allergic rhinitis (15.6%). Total IgE was not statistically significantly associated with clinical diagnosis(p = 0.889), age of the child (p = 0.102), gender (p = 0.687) or absolute eosinophil count (p = 0.318). The commonest allergens identified were dust mite (Dermatophygoides pteronissinus and D. farinae) and Bermuda grass. While antibody reaction to weeds, particularly plantain, were also common, these reactions were mostly mild to moderate. Allergy to cats and moulds was rare. CONCLUSION: In the absence of routine testing for specific allergens avoidance of dust mite and Bermuda grass seem important strategies in the management of difficult children with atopy. There is need for a prospective study to shed more light on the allergens that cause these common atopic conditions in our environment. PMID- 10695185 TI - The impact of an inadequate municipal water system on the residents of Chinhoyi town, Zimbabwe. AB - OBJECTIVE: To assess the use and impact of the water reticulation system in Chinhoyi on its residents. DESIGN: Cross sectional and case series studies. SETTING: Chinhoyi town. SUBJECTS: 600 Chinhoyi residents. MAIN OUTCOME MEASURES: Practices and perceptions of Chinhoyi residents on the water system, and distribution of water-related diseases per area of residence. RESULTS: Out of 600 respondents, 565 (99.3%) had access to piped water and 558 (98.0%) to flush toilets. Breakdowns of water supply and functioning of toilet facility were reported by 308 (77.0%) and 110 (28.0%) respondents in the previous six months, respectively. Main complaints of Chinhoyi residents were about low water quality (36.2%), inadequate sewage system (31.3%) and environmental pollution (26.5%). Cases of water-related diseases were not associated with natural water bodies. CONCLUSIONS: Chinhoyi residents have good access to the municipal water and an adequate sanitation system. However, low quality of the water, frequent system breakdowns and the degradation and loss of amenity of the environment impair their quality of life. PMID- 10695186 TI - Tuberculosis liver abscess in an HIV-infected patient. PMID- 10695187 TI - Hardly a harmless analgesic. PMID- 10695188 TI - The need for a regular comprehensive health surveillance on industrial workers. AB - The movement of people during some of their daily occupations and activities releases an amount of dust into the air. Even dust that has settled on floors and flat surfaces is made airborne by air currents. Of greater concern is the dusty environment caused by the operations within the workplace, such as handling of dusty materials, machining, cutting, drilling, milling, rock blastic and pounding. Fortunately, a lot of the dust is harmless except when present in high concentrations when it can cause some discomfort. At such levels it is termed 'nuisance dust'. However, some form of dusts are distinctly harmful, giving rise to impairment of lung function and pneumoconiosis (dust-induced changes in the lung). PMID- 10695189 TI - Oh no, not nitric oxide! AB - Nitric oxide (NO) is a unique, endogenous regulatory molecule that is involved in a wide variety of physiological processes in multiple organ systems. This simple gas functions as a cellular messenger in a broad range of biological activities that include blood pressure regulation, immunomodulation and neurotransmission. It has also been implicated in a number of homeostatic functions in the cardiovascular system: it is a significant determinant of basal vascular tone and, in addition is thought to regulate myocardial contractility and platelet aggregation. Dysregulation of NO mediated effects have been implicated in the pathogenesis of essential hypertension, atherosclerosis, and the hypotension associated with septic shock. This review will focus on these multiple effects of NO in the cardiovascular system. PMID- 10695190 TI - Hypotension and bedside leukocyte reduction filters. PMID- 10695191 TI - Uses and misuses of percentages. PMID- 10695192 TI - Risk factors for neonatal mortality: Harare Central Hospital Neonatal Unit- Zimbabwe. AB - OBJECTIVE: To assess risk factors for neonatal mortality in a tertiary level neonatal unit. DESIGN: Case control analysis of routine neonatal data for 1998. SETTING: Harare Central Hospital Neonatal Unit. SUBJECTS: All neonates delivered at Harare Maternity Hospital and admitted to the neonatal unit for care between January and December 1998. MAIN OUTCOME MEASURE: Neonatal mortality in hospital. RESULTS: A total of 5,305 neonatal admissions were studied of which 19.3% died in hospital. The median age at death was two days (Q1 = 1, Q3 = 3) and the median age at hospital discharge was 3 days (Q1 = 1, Q3 = 6). Risk factors for mortality were un-booked mother odds ratio (OR) 2.36 (95% CI = 1.98 to 2.81), breech delivery OR: 1.76 (95% CI = 1.39 to 2.22), low birth weight OR: 4.67 (95% CI = 3.92 to 5.57), prematurity OR: 2.36 (95% CI = 2.09 to 2.66), congenital malformations OR: 2.80 (95% CI = 1.72 to 4.53) and birth asphyxia OR: 1.79 (95% CI = 1.51-2.12). Being admitted for respiratory distress was associated with better survival OR: 0.22 (95% CI = 0.17 to 0.28). Having a Caesarian section was also protective OR: 0.60 (95% CI = 0.47 to 0.76). Mother's age, parity, time of delivery and sex were not significantly associated with mortality odds ratios (95% CI) of 1.07 (0.86 to 1.34), 0.94 (0.78 to 1.13), 1.10 (0.93 to 1.30) and 0.89 (0.78 to 1.03) respectively. On regression analysis birth weight greater than 2,500 g, being un booked and breech delivery were predictive of mortality with OR (95% CI) of 0.99 (0.99 to 0.99), 1.31 (1.12 to 1.61) and 1.15 (1.04 to 1.28) respectively. CONCLUSION: Low birth weight is the highest risk factor for mortality in this tertiary level hospital. Strategies targeted at low birth weight infants such as antenatal corticosteroid use, improved intrapartum care, appropriate antibiotic use, improved efficiency and access to neonatal intensive care will have the most impact on neonatal mortality. PMID- 10695193 TI - Highlights of extranodal lymphomas in Ibadan, Nigeria. AB - OBJECTIVE: To find out the pattern, trend and site distribution of extranodal lymphomas in Ibadan. DESIGN: Case series. SETTING: Department of Pathology, University College Hospital. SUBJECTS: All cases of malignant lymphomas with extranodal manifestations between 1981 and 1998. MAIN OUTCOME MEASURES: Frequency and distribution of primary extranodal lymphomas. RESULTS: Extranodal lymphomas constituted 9.8% of non-Hodgkin's lymphomas(NHLs) during the period, increasing from 8.5% of NHLs from 1980 to 1990 to 12.4% from 1991 to 1998. During the entire 18 year period the nasal/nasopharynx, gastro-intestinal tract (GIT), tonsils and skin were the most commonly affected sites constituting 20.2%, 19.3%, 14.3% and 13.4%, respectively of all extranodal lymphomas. However, between 1980 and 1991 the GIT was the most commonly affected site (26.1%) followed by the nasal/nasopharyngeal region (11.6%). The involvement of the nasopharyngeal area during the second period of the study accounted for 32.0%. CONCLUSION: The reasons for this changing trend are unknown and the significance of HIV infection in this regard is uncertain. PMID- 10695194 TI - Laparoscopic cholecystectomy in black patients at Ga-Rankuwa Hospital: a feasibility study. AB - OBJECTIVE: To determine feasibility and outcome of laparoscopic cholecystectomy for Black patients in Ga-Rankuwa Hospital and to analyse the type of stones harvested. DESIGN: Cross sectional description study. SETTING: Ga-Rankuwa Hospital/Medunsa, South Africa. SUBJECTS: 79 patients who presented with cholelithiasis over a six year period. MAIN OUTCOME MEASURES: Successful performance of laparoscopic cholecystectomy on the subjects. RESULTS: 39 laparoscopic cholecystectomies were performed with seven conversions (18% conversion rate). Forty open cholecystectomies were also performed. Post operative stay after laparoscopic cholecystectomy averaged 3.5 days and that of the open converted group 6.9 days (p < 0.001). Two patients returned with persistence of symptoms following laparoscopic cholecystectomy. Twenty gall stones were analysed; 17 were of the cholesterol type and three of the black pigment type. CONCLUSIONS: Laparoscopic cholecystectomy is feasible in the Black populace of Ga-Rankuwa Hospital. The majority of gallstones are of the cholesterol type. PMID- 10695195 TI - Beliefs about sexual relationships and behaviour among commercial farm residents in Zimbabwe. AB - OBJECTIVES: To describe beliefs of farm residents about sexual relationships and sexual behaviours within these relationships, as a basis for understanding how these beliefs may influence sexual behaviour; and to document gender differences in these beliefs. DESIGN: Cross sectional descriptive study. SETTING: Three commercial farms in Mashonaland Central Province, Zimbabwe. SUBJECTS: Purposive convenience sample of 218 adult (age 18+, or ever-married) residents on the commercial farms. MAIN OUTCOME MEASURES: Views towards celibacy, wife inheritance, condom use, infertility, extra-marital affairs. RESULTS: 85% of respondents felt marriage was a cushion against HIV/AIDS. The majority (87.6%) viewed wife inheritance as improper; 37.6% felt that it facilitated the spread of HIV. Sixty two percent viewed condom use in marriage as unacceptable, and the majority of respondents felt that infertility was a problem in a person's life, with men being less likely than women to indicate this (OR: 0.26; 95% CI: 0.10 to 0.67). Male respondents expressed greater ability than their female counterparts to take protective steps in the event of suspecting STD/HIV/AIDS in their spouse. More male than female respondents felt they could leave marriage against perceived risk of infection with HIV from their partner (OR: 2.20; 95% CI: 1.22 to 4.00). CONCLUSION: Perceptions of wife inheritance are positive when viewed against known ways of HIV transmission. There is need for further work to allow women to express their integrity as individuals in their relationship with men, as a strategy to promote their reproductive health. Determinants of condom acceptance need further understanding because condom use is one of the effective ways of curbing the spread of HIV/AIDS. PMID- 10695196 TI - Sporadic Peutz-Jeghers syndrome in a Nigerian. AB - Peutz-Jeghers syndrome is a rare condition of muco-cutaneous pigmentation and polyposis of the gut. Reports of its occurrence from the Black world have been infrequent. A case is presented of an 18 year old Nigerian girl with pigmentation of the inner lips and soles of both feet, and recurrent attacks of abdominal pain necessitating two surgical procedures for intestinal obstruction with removal, in both cases, of polyps. Problems of complications and therapeutic modalities are discussed. PMID- 10695197 TI - Images in cardiovascular medicine: ruptured aneurysm of the sinus of Valsava. AB - A 27 year old woman presented with a two year history of cardiac symptoms. Echocardiographic examination revealed a ruptured aneurysm of the sinus of Valsava. Of note the patient was known to be HIV positive. PMID- 10695198 TI - Why beta blockers should be used in heart failure. AB - Beta blockade may be beneficial for most patients with congestive cardiac failure (CCF) whether due to ischaemic or idiopathic dilated cardiomyopathy (DCM), although they are more effective if the CCF is idiopathic. Beta blockers are additive to angiotensin converting enzyme (ACE) inhibitors in their effects on CCF. Beta blockers have been shown in most studies to increase ejection fraction, cardiac output, and exercise capacity and are sometimes capable of resolving almost all the symptoms of heart failure. Treatment should begin with the smallest possible dose and this should be gradually increased to the maximum tolerated level. PMID- 10695199 TI - Informed consent for blood transfusion. PMID- 10695201 TI - Use of a medical decision support system to improve the preoperative diagnosis of prostate cancer with pelvic lymph node metastases. AB - BACKGROUND: We evaluated the effects of a medical decision support system on the preoperative diagnosis of prostate cancer with pelvic lymph node metastases. METHODS: The preoperative accuracy of staging prostate cancer with pelvic lymph node metastasis by the prostate cancer expert system (PCES) for 43 patients was compared to the accuracy of staging performed by 2 urological attending physicians and 5 residents, to test the validity of the PCES. The effect of PCES consultation on physicians' staging for prostate cancer with pelvic lymph node involvement was evaluated. RESULTS: In the diagnosis of prostate cancer with pelvic lymph node metastasis, PCES was significantly more accurate than the two attending physicians alone (p = 0.042; p = 0.008). All the urological residents' diagnoses were significantly less accurate than those of the PCES. After PCES consultation, all the urological residents increased diagnostic specificity significantly. Most residents usually used PCES for consultation only after the attending physician or department asked for the results. CONCLUSION: Owing to an increased ability for preoperative diagnosis of prostate cancer with pelvic lymph node metastasis, as supported by the PCES, some unnecessary pelvic lymphadenectomies may be avoided. PMID- 10695200 TI - Myocardial effects of beta-agonist stimulation in rats with chronic left ventricular dysfunction treated with an angiotensin-converting enzyme inhibitor. AB - BACKGROUND: This study measured morphological and hemodynamic changes and renin angiotensin responsiveness of the left ventricle (LV) to beta-agonist stimulation in a Sprague-Dawley rat model of myocardial dysfunction produced by coronary artery ligation. METHODS: The LV function and papillary muscle mechanics were measured after 12 weeks of captopril treatment (2 g/l in drinking water) following left coronary artery ligation or a sham operation. Fifty-two rats were divided into three groups: those with sham operations, those with small infarcts (infarct size [IS] < 30% LV) and those with large infarcts (IS > or = 30% LV). RESULTS: The results showed that LV end-diastolic pressures were elevated in the large-infarct group regardless of treatment with the angiotensin-converting enzyme inhibitor (ACEI), and the LV weight was reduced in the ACEI-treated rats. In addition, the uninfarcted LV posterior papillary muscle of the large-infarct rats showed an impaired response to isoproterenol stimulation, including the developed tension, positive and negative rate of tension development, time to peak tension, and time to half relaxation. CONCLUSION: Chronic captopril treatment improved isoproterenol-stimulated muscle isometric function in rats following myocardial infarction, possibly through the beta-receptor pathway. PMID- 10695202 TI - A comparative study of various fixation methods for mandibular fracture. AB - BACKGROUND: Internal rigid fixation for mandibular fractures has been recognized as a reliable method for obtaining osteosynthesis. It may allow for early active physiotherapy and resumption of normal function. However, few studies have compared the various fixation methods. METHODS: From January 1993 through December 1997, 101 patients with mandibular fractures, who were selected for study, received treatment using various fixation methods at the Craniofacial Center, Chang Gung Memorial Hospital. The fixation methods included plate fixation in 44 patients, lag screw fixation in 30, combined plate and lag screw fixation in 15, and wire fixation in 12. Clinical data assessment was performed by reviewing hospital records. For assessment of the long-term surgical results, the patients were asked to complete a questionnaire including questions which specifically targeted history regarding occlusion, mastication, facial asymmetry, width of mouth opening, and general appearance. RESULTS: The outcome assessment showed statistically different results among the groups. The wire group required intermaxillary fixation more often and for a longer duration compared to the groups using plates and/or lag screws. The plate, lag screw, and combined plate and lag screw fixation groups had better total outcome scores, in particular for long-term dental occlusion and mastication function. CONCLUSION: This study shows that for mandible fractures, the more rigid types of fixation methods, i.e., plates and screws or lag screws, can offer better short and long-term outcomes. PMID- 10695203 TI - Surgical outcome of combined phacoemulsification and trabeculectomy. AB - BACKGROUND: A retrospective study was undertaken to evaluate the results of phacoemulsification, intraocular lens implantation, and trabeculectomy in patients with cataracts coexisting with glaucoma. METHODS: This study consisted of 20 eyes from 19 patients. Fifteen eyes had chronic angle-closure glaucoma and 5 eyes had primary open-angle glaucoma. All cases were followed for a minimum of 6 months (range, 6 to 16 months). The mean preoperative intraocular pressure (IOP) was 17.3 +/- 4.5 mmHg. The mean preoperative visual acuity was 0.05 +/- 0.19. The mean number of preoperative antiglaucoma medications per patient was 2.3. RESULTS: Postoperatively, all patients except for one no longer required antiglaucoma medication. The mean IOP was 10.7 +/- 3.7 mmHg one month postoperatively and 13.7 +/- 4.3 mmHg at the final follow-up visit. Vision improved in 80% of the patients and remained unchanged in 20%. The failure to achieve improvement was due to advanced optic atrophy. The mean level of surgically induced astigmatism at the final visit was 0.98 +/- 0.91 diopters as calculated by vector analysis. Various extents of fibrin exudate was found in 10 eyes (50%). The most serious postoperative complication, occurring in one eye (5%), was temporary hypotony with moderate choroidal effusion, which later resolved spontaneously. CONCLUSION: Combined phacoemulsification and trabeculectomy is an effective and safe approach for obtaining good visual rehabilitation and glaucoma control. PMID- 10695204 TI - Clinical implication of atypical squamous cells of undetermined significance with or without favoring high-grade squamous intraepithelial lesion on cervical smears. AB - BACKGROUND: The cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS) on a cervical smear usually makes clinicians unsure of how to manage the patient and follow-up on her condition. We attempted to define the clinical implication of qualifying the cytologic diagnosis of ASCUS as either favoring a high-grade squamous intraepithelial lesion (HSIL) or not in an effort to provide management guidelines. METHODS: From January through May 1997, 65 of 5792 women who had cervical/vaginal smears taken at Kaohsiung Chang Gung Memorial Hospital were diagnosed as having ASCUS. Thirteen of the 65 cases of ASCUS favored an HSIL, based on nuclear abnormalities in atypical metaplastic and parakeratotic-type squamous cells. All these 65 patients were evaluated in our outpatient clinic by a second cervical smear, colposcopy, and colposcopically directed biopsies and/or endocervical curettage. The median length of the follow up period was 19 months (range, 16 to 21 months). RESULTS: Of the 52 patients evaluated for ASCUS smears without favoring HSIL, 6 (11.5%) had a low-grade SIL (LSIL), 1 (1.9%) had cervical intraepithelial neoplasia grade II (CIN II), and 1 (1.9%) had invasive squamous carcinoma. Of the 13 patients with a cervical cytologic diagnosis of ASCUS favoring HSIL, 1 (7.6%) had immature metaplasia, 2 (15.4%) had LSIL, 2 (15.4%) had CIN II, 6 (46.2%) had CIN III, and 2 (15.4%) had invasive squamous carcinoma. CONCLUSION: For patients with a cytologic diagnosis of ASCUS favoring HSIL, more aggressive interventions, such as colposcopy directed biopsy, endocervical curettage, or even conization, should be performed promptly. However, those without features favoring HSIL may be evaluated with regularly repeated smears. PMID- 10695205 TI - Laparoscopic colectomy is superior to laparotomy for reduction of disability in patients with colorectal adenoma. AB - BACKGROUND: This study was to evaluate disability after laparoscopic colectomy in patients with colorectal adenomas as compared to disability after laparotomy. METHODS: Patients who underwent laparoscopic colectomy for colorectal adenoma were compared to patients who underwent laparotomy for the same problem by the same surgeons during the same time period in Cleveland Clinic Florida. A standard questionnaire was used to assess disability which included the number of days to return to partial activity, full activity, and work. RESULTS: Twenty-nine patients who underwent laparoscopy were compared with 31 patients who underwent laparotomy. There were no significant differences in age (70.4 vs 72.5 years) (p = 0.405) or gender (M:F 13:16 vs 20:11) (p = 0.126) between the laparoscopy and laparotomy groups. The operative time was longer for the laparoscopy group than the laparotomy group: 170 vs 131 minutes (p = 0.014). However, the duration of postoperative ileus, hospitalization, time until return to partial activity, time until return to full activity, and time off of work were significantly shorter in the laparoscopy group than in the laparotomy group: 3.3 vs 5.2 days, 6.2 vs 8.7 days, 2.3 vs 4.2 weeks, 4.4 vs 9.3 weeks, and 3.7 vs 7.3 weeks, respectively (p < 0.041 for all). Although the incidence of postoperative complications was not significantly different (24% for laparoscopy vs 29% for laparotomy, p = 0.325), the incidence of postoperative prolonged ileus was statistically significantly lower in the laparoscopy group (3% vs 26%, p = 0.027). CONCLUSION: Laparoscopic colectomy for patients with colorectal adenoma can reduce postoperative ileus, postoperative hospitalization, and disability in terms of a quicker return to partial activity, full activity, and employment. Laparoscopic colectomy should be considered for all patients who have colorectal adenomas which require resection. PMID- 10695206 TI - Comparison of nasal trauma associated with nasopharyngeal airway applied by nurses and experienced anesthesiologists. AB - BACKGROUND: Insertion of a nasopharyngeal airway by nurses is considered to be invasive. We compared the incidence and severity of nasal injury associated with nasopharyngeal airway insertion by trained nurses to those by anesthesiologists to determine the safety of inserting a nasopharyngeal airway by nurses in cardiopulmonary resuscitation (CPR). METHODS: One hundred and sixteen male and 96 female patients scheduled for general anesthesia were included in the study. The male and female patients were randomly assigned to two groups respectively. Anesthesia was induced with sodium thiopental and fentanyl intravenously. The patients were then ventilated with a bag-valve-mask by trained nurses or anesthesiologists. In the unsuccessfully ventilated patients, nasopharyngeal airways were inserted to facilitate subsequent ventilation. The nasopharyngeal airway, oropharynx, and nostrils were then examined. The incidence and severity of nasal trauma induced by trained nurses or by anesthesiologists were compared. RESULTS: The study revealed that nasopharyngeal airways applied by trained nurses did not induce more severe nasal trauma than those by anesthesiologists. CONCLUSION: We suggest that nasopharyngeal airways may be applied safely by trained nurses in CPR. PMID- 10695207 TI - Enterovesical fistula: experiences with 41 cases in 12 years. AB - BACKGROUND: A retrospective analysis of enterovesical fistula treated at Chang Gung Memorial Hospital was conducted to determine the optimal diagnosis and management of this disease. METHODS: The records of 41 patients who presented from 1984 to 1996 and had a final diagnosis of enterovesical fistula were retrospectively reviewed. The etiology, symptoms on presentation, diagnostic tools, and modality of treatment were analyzed. RESULTS: The majority of these cases were associated with malignancy (38, 92.7%), and the others with diverticulitis (2, 4.9%) and iatrogenic causes (1, 2.4%). In those with malignancy, 15 patients (39.5%) were found to have tumor recurrence. The most frequent symptom in enterovesical fistula was fecaluria (58.5%), followed by abdominal pain (22%) and dysuria (14.6%). Diagnostic tools included the barium enema, cystography, and cystoscopy; these methods could identify the fistula in 63.2%, 60%, and 53.8% of the patients, respectively. Methods of management included diversion only (39%), one-stage fistula repair (36.6%), and watchful surveillance (24.4%). CONCLUSION: Enterovesical fistula should be considered if fecaluria, pneumaturia, or persistent non-specific urinary tract infection present as the initial complaint. A thorough surgery for a possible underlying malignancy is mandatory when confronted with enterovesical fistula, since the incidence of inflammatory bowel disease is low in this area. An abdominal computer tomography (CT) scan, barium enema, and cystogram can be useful diagnostic tools. Treatment of this entity should be individualized according to each patients clinical status. PMID- 10695208 TI - Anterior knee pain after intramedullary tibial nailing. AB - BACKGROUND: Currently, intramedullary nailing is a well-accepted method for treating tibial shaft fractures, but some patients complain of anterior knee pain after surgery. Multiple factors may influence this troublesome complication. METHODS: This was a retrospective analysis of the medical records of 200 patients who were treated with intramedullary nailing after tibial shaft fractures. Sixty four patients complained of knee pain after surgery. We evaluated the knee pain in relation to the surgical approach, radiographic readings, and the type of nail used. RESULTS: Among the 64 patients, 45 (70%) received central approaches and 19 (30%) received paramedial approaches (p = 0.0002); 46 patients (72%) showed nail protrusions on radiographs and only 18 patients (28%) were without nail impingement (p = 0.0001). Forty-three patients (67%) received Kuntscher nail fixation and just 21 patients (33%) had interlocking nail fixation (p = 0.0015). CONCLUSION: The use of the central patellar tendon splitted approach, nail protrusion observed on radiographs, and the insertion of a non-locking intramedullary nail were all significant risk factors for anterior knee pain after surgery. All these risk factors should be avoided in tibial nailing to decrease the problem of postoperative knee pain. PMID- 10695209 TI - Evaluation of peribulbar anesthesia in encircling scleral buckle surgery and its postoperative pain course. AB - BACKGROUND: Retrobulbar anesthesia is considered effective in ocular surgery but it can give rise to serious complications. We used peribulbar anesthesia with sub Tenon's irrigation to perform encircling scleral buckling for retinal detachment, as it could reduce the complications caused by retrobulbar anesthesia. We also recorded the course of pain for 72 hours after surgery. METHODS: Thirty patients who were diagnosed with rhegmatogenous retinal detachment were treated with an encircling scleral buckle. The surgery was performed with peribulbar anesthesia with occasional sub-Tenon's irrigation. We evaluated the patient's pain with a visual analogue scale after surgery at 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, and 72 hours. RESULTS: In 24 cases (80%), the anesthesia was complete with the peribulbar block. Only 6 patients (20%) needed sub-Tenon's irrigation and four of them felt no pain after augmentation. Although all the surgical procedures proceeded without problem, two of the patients felt pain and were uncomfortable during the surgery. No serious complications occurred. The course of pain peaked 6 hours after surgery when 26 patients (86.7%) felt pain and 12 patients (40%) were uncomfortable (pain score > or = 5). Forty-eight hours after surgery, 9 patients (30%) still felt pain but no one felt uncomfortable. CONCLUSION: Peribulbar anesthesia can be used safely in encircling scleral buckling for retinal detachment. The postoperative pain is maximal 6 hours after surgery and becomes mild (pain score < or = 4) after 48 hours. PMID- 10695210 TI - Neonatal vallecular cyst: report of eleven cases. AB - BACKGROUND: Vallecular cyst is fairly uncommon in neonates and infants. Although benign in nature, it may cause severe airway obstruction and even death. This study retrospectively analyzed the clinical manifestations of vallecular cyst in neonates and discussed its management. METHODS: From June 1993 through January 1997, 11 cases of vallecular cyst were collected and reviewed retrospectively. There were 8 male infants and 3 female infants. Their clinical manifestations, age at the onset of symptoms, age at diagnosis, and surgical management were analyzed. Fibrolaryngoscopy was used for preoperative diagnosis and postoperative follow-up. RESULTS: The infants' initial presentations were inspiratory stridor, respiratory distress, noisy breathing, feeding difficulty, and failure to thrive. There was a high incidence of patients with coexisting signs of laryngomalacia (10/11). Ten patients received laryngomicrosurgery with CO2 laser for deroofing the cyst. Additional supraglottoplasty was performed at the same time in 4 patients with laryngomalacia type A + C and in one patient with severe laryngomalacia type C. Their symptoms all resolved soon after surgery. The phenomenon of laryngomalacia also disappeared. There has been no recurrence up to the present. CONCLUSION: Although fairly uncommon, vallecular cyst should be included in the differential diagnosis of congenital laryngeal stridor in neonates. The use of fibroendoscopy can promptly diagnose vallecular cyst and any synchronous airway lesions. Although most of the synchronous laryngomalacia (type C) in this study was secondary to vallecular cyst, we suggest that supraglottoplasty be taken into consideration during cyst deroofing when the signs and symptoms of laryngomalacia type A are also present. PMID- 10695211 TI - Renal autotransplantation for ureter stricture and renovascular disorders. AB - BACKGROUND: Renal autotransplantation is an established therapy in cases of renal vascular lesions, tumors of the kidney and ureter, complex ureteral lesions, and kidney trauma. It has been a significant technical innovation, aiding the urologist in his great effort to preserve renal function by conserving renal tissue. We report our experience with autotransplantation in 4 patients. The indications, techniques, and results of renal autotransplantation in relation to our own experience are discussed. METHODS: The patients included 3 women and one man. The average age of the patients was 35 years old, with a range from 20 to 54 years. One patient had Takayasu's arteritis, the second had Nutcracker syndrome with flank pain and hematuria, the third a complicated long ureter stricture, and the fourth patient a renal artery saccular aneurysm. RESULTS: The average operation time was 7 hours (4.5 to 8.5 hours), and the cold ischemia time was about 88 minutes (45 to 150 minutes). Three of the autografts resumed normal renal perfusion, and in the fourth patient the renal autograft was lost due to vascular thrombosis. CONCLUSION: Renal autotransplantation is a feasible method for the surgical treatment of renal and ureteral lesions. To avoid postoperative ureteral sloughing and subsequent urinary fistulas, the ureter can be left intact to preserve the ureter blood supply. However, in the case of a complicated vascular reconstruction procedure, it appears to be appropriate to divide the ureter and have the kidney completely free, thus avoiding back-flow perfusion from the intrinsic and intercommunicating blood supply in the ureteral wall, which may result in vascular thrombosis and subsequent autograft failure. PMID- 10695212 TI - Usefulness of pediatric flexible bronchoscopy in the early diagnosis and postoperation evaluation of vascular rings: report of three cases. AB - Vascular rings are a diagnostic and therapeutic challenge for pediatricians. Many diagnostic modalities contribute to the detection of these rare congenital vascular anomalies. The role of flexible bronchoscopy is still being debated. We present 3 cases to emphasize the usefulness of pediatric flexible bronchoscopy (PFB) in the early diagnosis and postoperative evaluation of vascular rings. In patient 1, PFB was performed before conventional techniques were available. A right aortic arch with a retroesophageal aortic diverticulum and mirror-image branching was later confirmed. In patients 2 and 3, pulmonary artery slings could not be detected clearly by imaging studies before PFB was performed. PFB was arranged again postoperatively for these 2 patients, because of difficulty weaning patient 2 from ventilator support and persistent respiratory distress in patient 3. In conclusion, we expect that more vascular rings will be diagnosed using PFB. This instrument is also useful in making a decision for surgery, and for detecting associated tracheobronchial anomalies preoperatively, assuring appropriate correction intraoperatively, and monitoring the condition of vascular rings postoperatively. PMID- 10695213 TI - A new technique for generating a computer-aided design and computer-integrated machining crown: case report. AB - A new technique for producing a Computer-Aided Design and Computer-Integrated Machining Ceramic Reconstruction crown is presented. After completion of the tooth preparation, an "optical" impression of the tooth was made with a charged couple device camera, and the electronic image was transferred to a computer screen. The "proposed" crown was electronically designed on the screen by the operator, and a proper prefabricated ceramic block was selected and used as milling material. A miniature milling machine then fabricated the crown from the ceramic block. The marginal adaptation and the contour of the crown were verified, and an external shading technique was utilized to improve the esthetics. The prepared tooth and crown were acid-etched, and the crown was cemented with a dual-curing composite luting agent. Once bonded in place, the occlusion was adjusted and the crown polished and finished. The advantage of this technique is that it eliminates the traditional laboratory casting procedure and corresponding laboratory fee while utilizing materials with superior physical properties for maximum strength and esthetics. PMID- 10695214 TI - A huge 6.2 kilogram uterine myoma coinciding with omental leiomyosarcoma: case report. AB - Surgery for massive abdominal tumors is both interesting and challenging. We present a case involving a multiple uterine myoma weighing 6.2 Kg which coincided with omental leiomyosarcoma. To our knowledge, this is the first report of this type of condition in the English literature. A 44-year-old nulliparous woman had suffered from abdominal pain for a long time. A huge abdominal mass was palpated on physical examination. Computed tomography scanning revealed a huge pelvic abdominal mass with the possibility of small bowel loops invaded by the mass. A 6 cm omental mass was incidentally found during the subsequent hysterectomy procedure. Perforation of the urinary bladder occurred during the dissection of adhesion. Resection of the omental mass, wide wedge resection of the invaded small bowel, primary repair of the bladder, and hysterectomy were performed. The final pathologic diagnosis was uterine leiomyomata with omental leiomyosarcoma. The patient returned home on postoperative day 14 and was well at the 18-month follow-up examination. The challenge of these tumors lies in their proper diagnosis and surgical management. More case reports and follow-up studies are needed to confirm the efficacy of their management. PMID- 10695215 TI - Octreotide treatment for a malignant islet cell tumor with variable hormone secretion: case report. AB - It is well known that an islet cell tumor can secrete multiple hormones depending on its cell type. We report the case of a 70-year-old woman who initially presented with peptic ulcer symptoms, an elevated serum gastrin level, and multiple liver tumors. Liver biopsy and distal pancreatectomy were performed, and the pathological diagnosis was malignant islet cell tumor. Additionally, the immunohistochemical staining revealed scattered positivity for gastrin, and then also positivity for insulin 14 months later. A subsequent hypoglycemic episode and elevated serum gastrin and insulin levels suggested that the disease had developed into a condition of multiple hormone secretion. The plasma gastrin and insulin levels decreased from 584 pg/ml and 90.8 microIU/ml to 49.1 pg/ml and 20.9 microIU/ml, respectively, 5 days after treatment with subcutaneous octreotide 100 micrograms every 6 to 8 hours. In addition, follow-up computed tomography showed shrinkage of the metastatic liver tumors. In conclusion, we found a case of malignant islet cell tumor with variable hormone secretion which could be effectively controlled with octreotide. PMID- 10695216 TI - Adult Citrobacter freundii meningitis: case report. AB - Citrobacter is a distinct group of Gram-negative bacilli belonging to the Enterobacteriaceae family. Central nervous system (CNS) infections due to Citrobacter are uncommon, though they occur more frequently in neonates and young children. In adults, Citrobacter meningitis is extremely unusual with only 6 cases reported in the literature before 1998. This rare CNS infection has been seen in patients with head trauma, following neurosurgical procedures, and in those who are immunocompromised. Of the patients in the 6 reported cases, only one developed multi-antibiotic resistant Citrobacter CNS infection. Adding to this small number of reported cases, we report an adult case of post neurosurgical meningitis and subdural empyema caused by multi-antibiotic resistant Citrobacter freundii and also review the literature related to this infection. Antimicrobial therapy with imipenem and third-generation cephalosporins failed to result in cerebrospinal fluid sterilization in our patient. Because of the use of broad-spectrum antibiotics, multi-antibiotic resistant Citrobacter species have developed in this nosocomial CNS infection and now present a therapeutic challenge. Therefore, further clinical studies are needed to determine updated therapeutic modalities for treating this life threatening infection. PMID- 10695217 TI - Radical resection and intraoperative radiotherapy for a recurrent endometrial cancer after prolonged remission following aggressive salvage therapy: case report. AB - The prognosis of recurrent endometrial carcinomas is generally poor, except for isolated vaginal relapse. We report a case of recurrent endometrial cancer in a 58-year-old woman who initially received a type I extended hysterectomy with bilateral salpin-go-oophorectomy and bilateral para-aortic and pelvic lymph node dissection. The first recurrence occurred in the left parametrium 7 months after the primary surgery. The salvage therapy consisted of radiotherapy combined with hormonal therapy (tamoxifen and Megace). Complete remission was achieved initially. Subsequently, the patient accepted six courses of chemotherapy (cisplatin and Adriamycin) for progressive elevation of cancer antigen 125 (CA 125). The CA-125 levels remained elevated with titers fluctuating around 100 U/ml until a second recurrence at the left iliac 75 months following salvage therapy. The second salvage treatment consisted of maximal debulking of the pelvic mass and intraoperative radiotherapy (IORT) followed by four courses of chemotherapy with paclitaxel and carboplatin. Complete remission was again accomplished, with clinical investigations and molecular markers returning to normal. The patient has been clinically free of disease for more than 2 years since the second relapse of cancer. In this particular case, we found that repeated recurrence could occur after a long complete remission following salvage therapy; however, the disease could be recontrolled with further aggressive salvage efforts. A multimodality approach with combinations of radical resection, IORT, and paclitaxel-based chemotherapy can be offered to patients with localized recurrent or repeatedly recurrent endometrial carcinoma after previous cisplatin-based chemotherapy and pelvic radiation. PMID- 10695218 TI - Alien hand syndrome: report of two cases. AB - Alien hand syndrome (AHS) refers to the occurrence of apparently purposeful movements in the hand which are independent of volitional control. Two subtypes of AHS have been proposed: frontal AHS, with grasp reflex and compulsive manipulation of tools by the dominant hand, and callosal AHS, which occurs in the nondominant hand and is characterized mainly by intermanual conflict. Here, we report two cases of frontal-type alien hand syndrome with symptoms of reflexive grasping, impulsive groping, and apraxia (in case 1), and compulsive manipulation of tools (in case 2). Brain computed tomography revealed a left anterior cerebral artery (ACA) territory infarct and multiple small infarcts of both hemispheres in patient 1 and a left ACA infarct in patient 2. The involuntary movements were bothersome to these patients in their daily activities. Both patients attended conventional physical and occupational therapies, and patient 2 received additional biofeedback training. Follow-up studies showed the spontaneous grasping behavior was still present in patient 1 and AHS had subsided in patient 2. We also describe a potentially effective technique involving biofeedback for patients with alien hand syndrome. PMID- 10695219 TI - Blunt trauma-induced internal thoracic artery injury: case report. AB - We report the case of a 54-year-old male motorcyclist with an apparent grade IV liver injury and life-threatening hemomediastinum and right hemothorax following blunt deceleration trauma. Massive hemothorax and an unstable hemodynamic status even under copious blood volume replacement made emergent surgical intervention mandatory. A midline laparotomy was performed at first to rule out abdominal bleeding accompanied by a diaphragmatic tear, but the procedure was soon converted to a thoracotomy after finding an intact diaphragm and persistent bleeding from the chest tube. An isolated internal thoracic artery (ITA) transection was identified. It was actively bleeding and causing a huge anterior mediastinal hematoma and had ruptured into the right pleural cavity. The bleeder was controlled with suture ligation and the hemodynamic status was soon stabilized. The patient recovered without significant sequelae. The rarity of this kind of presentation is discussed, including both the ITA injury mechanism and the problems posed in making an early and correct diagnosis. PMID- 10695220 TI - Unusual dilatation of Virchow-Robin spaces: case report. AB - Virchow-Robin spaces normally surround the perforating arteries that enter the brain. These spaces are a well-defined sites where immunological reactions take place and they may have implications in the pathogenesis of a number of neuropathological conditions. We present the case of a 52-year-old woman who had a history of complex partial seizures for 30 years. Her routine neurological examinations and mini-mental tests had normal results. Magnetic resonance images of this patient revealed unusual widening of the Virchow-Robin spaces up to 1.5 cm in diameter along the perforating medullary arteries in the white matter, more so in the left hemisphere. Although it has been concluded that these large spaces are a phenomenon of the normal aging brain and are unrelated to neurological diseases, our patient had had epileptic seizures for 30 years. The large Virchow Robin spaces of our patient might have been an incidental radiologic finding. Their pathogenesis remains unclear, and their possible clinical relationship to epilepsy deserves further pathological studies. PMID- 10695221 TI - Schistosoma japonicum infection presenting with colon perforation: case report. AB - Colon perforation can be caused by a variety of entities, including iatrogenic trauma, tumors, ischemia, inflammatory bowel disease, and steroid use. Parasitic infection rarely leads to colon perforation. Secondary peritonitis results from mixed microorganism infection, including enterococci, enteric bacilli, and anaerobes. A combination of an optimal antibiotic regimen and surgical intervention is of paramount importance. Nevertheless, intra-abdominal infections usually have a high mortality rate. Schistosomiasis occurs worldwide. S. japonicum infection is endemic in Asia. The most common complications of gastrointestinal schistosomiasis are periportal fibrosis, intestinal polyposis, and bowel stricture. Rarely, schistosomiasis results in colon perforation. The diagnosis of schistosome infections is based on ova in stool or tissue specimens, and/or immunologic diagnostic tests. The most effective anti-schistosomiasis agent is praziquantel. Herein, we describe an unusual case of colon perforation associated with Schistosoma japonicum infection, which resulted in severe peritonitis and led to the patient's death. PMID- 10695222 TI - Airway obstruction in general anesthesia--two different episodes in the same patient: case report. AB - The case of a patient with Apert's syndrome (acrocephalosyndactyly) who had a tracheostomy tube and who encountered two different episodes of critical airway obstruction during two different general-anesthetic procedures for craniofacial surgery is reported. The first episode, at the age of four, involved occlusion of the uncuffed tracheostomy tube by a blood clot, which might have come from the surgical field of the maxillary Le-Fort III advancement procedure. The second episode was encountered during his emergence from the general anesthesia of a degloving midface osteoplasty and a maxillary Le-Fort I osteotomy procedure 3 years later. Although a cuffed armored tube had been inserted through the tracheostoma to prevent aspiration of blood from the surgical field, the armored tube was plugged by a piece of granulation tissue that might have been dislodged from the peri-stomal area. Factors that lead to tracheostomy tube obstruction, their clinical features and preventive measures are discussed. We believe that being alert to changes in the airway pressure, the ventilation pattern, and the hemodynamic status is necessary during the administration of general anesthesia. Precautions should be taken at all times, particularly for patients with a tracheostomy. PMID- 10695223 TI - Hemorrhagic gastric glomus tumor mimicks a leiomyosarcoma on both transabdominal and endoscopic ultrasonography: case report. AB - Glomus tumor is a benign tumor that arises from the modified smooth muscle cells of the glomus body and usually occurs in the skin, particularly in the nail-beds and fingertips. Gastric glomus tumor is a rare gastric submucosal tumor. Endoscopic ultrasonography (EUS) is useful in differentiating the gastric submucosal tumors, such as leiomyoma, leiomyosarcoma, lipoma, ectopic pancreas and glomus tumor. On sonography, gastric glomus tumor appears as a homogeneous hypoechoic or a heterogeneous low echoic pattern mixed with internal high echoic spots. Here, we describe an unusual sonographic figure of a hemorrhagic gastric glomus tumor on both EUS and high-resolution transabdominal ultrasonography (TAU). This tumor was located in the gastric muscular layer. Sonographic examination revealed a heterogeneous echogenicity with hyper- and anechoic areas, which mimicked the echoic pattern of gastric leiomyosarcoma. PMID- 10695224 TI - The socioeconomic burden of hospital acquired infections. PMID- 10695225 TI - An analysis of 60 cases of culture bound syndromes. AB - Of 60 cases of culture bound syndromes seen in psychiatry OPD, Dhat syndrome was most common (76.7%), followed by possession syndrome (13.3%). Depression by the most common associated psychiatric disorder. As the data on culture bound syndromes in Indian subcontinent is less, this study calls for careful evaluation of these patients psychologically to detect and treat the associated psychiatric comorbidity appropriately. PMID- 10695226 TI - Experience with quinine in falciparum malaria. AB - During a period of 1 year from July 95 to June 96, 60 patients with falciparum malaria were treated with quinine at Kasturba Medical College Hospital, Mangalore. Of these, 24 patients developed adverse effects to quinine. They were cinchonism (15) cardiotoxicity (10) hypoglycemia (9) hyperventilation (3) hypersensitivity reactions (3) and hypokalemia (1). Cardiotoxicity was noted in 4 of the 7 patients who received intravenous quinine and all four had renal and hepatic failure and prolonged Q-Tc on electrocardiogram. All 4 died of cardiac arrhythmias, 2 had broad QRS tachycardia and 2 had sinus bradycardia. We conclude that: 1. Quinine should be used cautiously in patients with impaired hepatic or renal function and in those with prolonged QTc as it can lead to cardiotoxicity in the form of I0 AV block, prolonged Q-Tc, broad QRS tachycardai or fatal bradyarrhythmia. Dosage reduction to 5 mg/kg body weight in the patients seem to be safer. 2. Hypoglycemia is a very frequent complication of quinine therapy and special care and frequent blood sugar estimations are required especially if the patient has vomiting. 3. Parenteral quinine is more likely to cause toxicity than oral quinine as earlier described. PMID- 10695227 TI - A study of serum bilirubin in neonates in relation to the maternal age. AB - A comparative study of neonatal serum bilirubin levels was done in neonates of different age groups of mothers. A total 122 healthy, new borns were selected for the study, born at Queen Mary's Hospital, Lucknow. Mothers were divided into two groups i.e. < 30 years and > 30 years of age. Samples of blood were collected thrice, first on day 1 from cord blood, 2nd and 3rd on days three and five of life from neonates by heel prick method, using small bore capillaries for blood collection, serum bilirubin estimation were done by the method of Malloy & Evelyn and Mean +/- SD were calculated. P-Value was observed between different age groups. In both the groups of mothers i.e. < 30 years and > 30 years serum bilirubin levels in their neonates raised to highly significant levels on day 3 (P-Value < 0.001) as compared to their cord blood serum bilirubin levels. On comparing serum bilirubin levels in neonates of both the maternal groups, it was observed that there is no significant difference between two groups on day of birth and day day 5 but statistically significant difference was observed on day 3 (P < 0.05), serum bilirubin levels were higher in neonates of younger age group mothers. PMID- 10695228 TI - Sociomedical problems of institutionalised women. AB - The inmates of this institute form a special group in the society which is a reflection of various social pressures and problems. These institutionalised female largely come from poor socioeconomic families and often from families with serious social problems like family disharmony (30%), marital disharmony (20%), poverty (10%), unwedmotherhood (8%), broken homes (51.1%) etc. 90% inmates had one or more morbid conditions. Average number of morbid conditions per inmate was 2.16 at the time of study. Institution which assumes the custody of these women should provide them with medical services towards the rehabilitation and discharge of a healthy, productive, well adjusted citizens. An effective health education programme comprising of nutrition education, sex education, personal hygiene, alcohol and drug abuse should be specifically drafted and should be undertaken for the inmates. PMID- 10695229 TI - Knowledge and practices about HIV transmission among barbers of Nagpur City. AB - The present study was carried out in south zone of Nagpur city to assess knowledge and practices regarding HIV transmission of 375 barbers selected randomly from three different categories of saloons. A significantly large proportion of the roadside barbers were ignorant about modes of transmission of HIV, particularly through the blades. The practices observed by barbers ae found to be favourable for transmission of HIV, more so in roadside barbers. Hence it can be concluded that practices observed by barbers may favour transmission of HIV and there is a scope for educational intervention. PMID- 10695230 TI - Dynamics of contraceptive use in a rural community of Haryana. AB - A cross-sectional survey was carried out to study the dynamics of contraceptive use in three villages of Raipur Rani block in district Panchkula, Haryana. A female social worker interviewed 600 ever-married women aged 15-44 years using a semi-structured interview schedule. More than 75% of the respondents were aware about modern contraceptives. Fifty-nine percent of the couples had used contraceptives. Among the 351 ever-users, the first contraceptive method used was sterilization in 41.3%, condom in 35.6%, IUD in 17.9%, and oral pills in 5.1%. Subsequently, many of them either discontinued or shifted to other methods. At the time of survey, 236 (39.3%) were using a contraceptive. Most of the current users (225) had opted for tubectomy, and only a few (4) had accepted vasectomy. Supplies of contraceptives were obtained mainly from government hospitals and sub health centres. Common reasons for discontinuation were perceived untoward effects (37.1%), desire for more children (32.6%), and failure of the contraceptive method (19.0%). Most of the respondents obtained contraceptives from Government health posts. Counselling and follow-up services should should be strengthened so that contraceptives are used regularly and effectively for longer periods. PMID- 10695231 TI - Delayed immunization against vaccine preventable diseases--factors responsible among children under 5 years of age. AB - In the present case-control study, out of the the eleven risk factors of delayed immunization, only seven, namely family size, sex, number of children < 5 years, material education, paternal education, distance from health centre and low socio economic status were found to be significantly associated. The common causes for delayed immunization were negligence on part of parents, unawareness about the use of vaccine and sickness of child. Thus, health education of the parents is recommended. PMID- 10695232 TI - Bacteriological analysis of burn sepsis. AB - A total of 114 opportunistic bacteria were isolated from 65 swabs from burn sepsis. P. aeruginosa (53.8%) was the most common agent followed by S. aureus (38.4%), Klebsiella Spp. (27.6%), Proteus (18.4%), E. coil (10.7%) and others. The infection was monobacterial in 25 cases (38.4%) and polybacterial in 49 cases (61.5%). P. aeruginosa was predominated in both monobacterial and polybacterial infections. Ciprofloxacin (42.9%) was found to be the most effective antibacterial agent. Results indicate that resistance in burn isolates is higher and increasing day by day. PMID- 10695233 TI - Management of drug resistant tuberculosis. PMID- 10695234 TI - Importance of autopsy in prevention of epidemics. AB - With resurgence of infectious diseases all over the globe, there is need for constant surveillance and specially trained staff to expertly carry out autopsies on suspected cases and determine the exact cause of death. Early and accurate diagnosis is important in order to prevent worldwide spread of the disease. PMID- 10695235 TI - Using antileukotrienes in asthma therapy. PMID- 10695236 TI - Heart drug with 'viagra side-effect'. PMID- 10695237 TI - Alcohol drinking patterns and medical care use in an HMO setting. AB - The objective of this study was to examine the association of medical care use (outpatient visits and hospitalization) with alcohol drinking patterns in a large health maintenance organization (HMO). Data were gathered from a random sample of 10,292 adult respondents through a telephone survey conducted between June 1994 and February 1996. Findings indicate that current nondrinkers with no past history of drinking had higher rates of outpatient visits and hospitalizations than current drinkers. Among current drinkers, medical care use declined slightly as drinking levels increased. Among nondrinkers, those with a drinking history exhibited significantly higher use of outpatient visits and hospital care than nondrinkers with no drinking history and current drinkers. Controlling for demographic and socioeconomic factors, health status, and common medical conditions in multivariate analyses suggests that nondrinkers with a drinking history use more services because they are sicker than other nondrinkers or current drinkers. PMID- 10695238 TI - Behavioral health funding for Native Americans in Arizona: policy implications for states and tribes. AB - This article examines the principal structures and mechanisms used by federal and state government to fund the behavioral health needs of Native American Indians. Using Arizona as a case study, the article provides an overview of both federal and state programs, especially Medicaid, discussing the problems and strengths of each. The article concludes with a discussion of the policy implications of these programs for both states and tribes, focusing on issues concerning administrative complexity, tribal sovereignty, improving behavioral health services, and assignment of financial risk. PMID- 10695240 TI - Predicting service utilization with the Child and Adolescent Functional Assessment Scale in a sample of youths with serious emotional disturbance served by center for mental health services-funded demonstrations. AB - This study investigated level of restrictiveness of living arrangements and number of days in out-of-family care at six months postintake, based on the Child and Adolescent Functional Assessment Scale (CAFAS), the Child Behavior Checklist (CBCL), gender, age, and level of family income at intake. It was composed of youths who met the criteria for serious emotional disturbance (SED) and were for the most part living in families that are described as socioeconomically disadvantaged. A multinomial logit model was used in the analysis of level of restrictiveness of living arrangements, and an ordinary least squares (OLS) regression model was conducted on number of days in out-of-family care. The CAFAS score at intake was found to be a significant predictor of service utilization between intake and six months and was a more consistent predictor than the CBCL. Results suggest that the CAFAS can be used to match service needs with resource allocation and to monitor performance-based outcome indicators. PMID- 10695239 TI - Comparing provider perceptions of access and utilization management in full-risk and no-risk Medicaid programs for adults with serious mental illness. AB - This article compares provider perceptions of access to services and utilization management (UM) procedures in two Medicaid programs in the same state: a full risk capitated managed care (MC) program and a no-risk, fee-for-service (FFS) program. Survey data were obtained from 198 mental health clinicians and administrators. The only difference found between respondents in the FFS and MC sites was that outpatient providers in the MC site reported significantly lower levels of access to high-intensity services than did providers in the FFS site (p < .001). Respondents in the two sites reported similar attitudes toward UM procedures, including a strong preference for internal over external UM procedures. These findings support the conclusion that through diffusion of UM procedures, all care in the Medicaid program for persons with a serious mental illness is managed, regardless of risk arrangement. Implications for mental health services and further research are discussed. PMID- 10695241 TI - Motivational versus confrontational interviewing: a comparison of substance abuse assessment practices at employee assistance programs. AB - The aim of this study was to conduct a quasi-experimental comparison of two employee assistance program (EAP) assessment approaches with substance abusers: confrontational interviewing (CI) and motivational interviewing (MI). A total of 176 EAP clients from 14 study sites met the study criteria, and 89 (51%) agreed to participate in the study. At three and nine months postassessment, both the MI and CI groups showed similar changes in readiness for change, completion of initial treatment plans, and subsequent treatment. Most important, both the MI and CI participants showed significant and comparable improvement on all of the substance abuse baseline measures as well as measures of family-social well-being and effects of drinking/drugging on work performance. The results open the door for EAP counselors to use an empirically supported assessment style that is at least as effective as the traditional confrontational approach. PMID- 10695242 TI - Psychometric evaluation of an inpatient psychiatric care consumer satisfaction survey. AB - This study assessed the psychometric properties of a questionnaire designed to measure consumer satisfaction with inpatient psychiatric care. To this end, 37 inpatient psychiatric units from across the United States agreed to participate. The questionnaire was completed by 1,351 individuals, or a responsible party, for an average response rate of 53%. The factor analysis identified six scales: Nonclinical Services, Psychiatric Care, Staff, Medical Outcome, Patient Education, and Program Components/Activities. The internal reliability of the scales was high to moderate (.88 to .74). Results of a stepwise regression model showed good criterion-related validity, explaining 58% of the variance in overall quality ratings. Little shrinkage in this variance occurred when the model was cross-validated. Also, differences in satisfaction levels were noted for select facility and consumer characteristics. Results are interpreted as providing support for the reliability and validity of a newly developed consumer satisfaction survey for use in evaluating inpatient psychiatric care. PMID- 10695243 TI - Distributive justice in Medicaid capitation: the evidence from Colorado. AB - In 1995, the state of Colorado began a new funding program for the provision of mental health services to Medicaid recipients. Medicaid funding was restructured from a fee-for-service system into a capitated managed care system. The restructuring altered the way in which mental health resources were allocated within Colorado's mental health system. This article explores the ethical principles inherent in the allocation of mental health resources within Colorado. The allocation system before and after the capitation pilot is analyzed according to three models of distributive justice. Under capitation, access to care corresponds to egalitarian ideals, while service delivery and outcomes follow a more utilitarian philosophy. Results from several empirical studies of the Colorado Medicaid system are used to support this ethical analysis. The analysis leads to the suggestion that the fair-opportunity rule may be a useful principle for developing just distribution systems in other states in the future. PMID- 10695244 TI - Cross-system service use among psychiatric patients: data from the Department of Veterans Affairs. AB - This study examines the cross-system use of non-Department of Veterans Affairs (VA) services in a sample of psychiatric patients from the VA in 1990. Data were collected over a two-week period on all mental health outpatients and included demographic information, diagnoses, and self-reported use of non-VA services in the previous two weeks and six months. In the entire sample, 10.6% and 23.3% reported cross-system use in the previous two weeks and six months, respectively. Predictors of cross-system use were lower VA utilization, a nonschizophrenic diagnosis, not having a VA service-connected disability, and being female. These data indicate that a substantial proportion of VA mental health patients are using non-VA services. Utilization patterns indicate that they may be substituting non-VA for VA services. These results are unlikely to be unique to VA, and rates of cross-system use will likely increase in all health care systems as financial restrictions increase. PMID- 10695245 TI - Linking substance abuse and serious mental illness service delivery systems: initiating a statewide collaborative. AB - In the past, persons with serious mental illness and substance abuse often found themselves in parallel systems of care that inadequately addressed their needs. Recent advances have seen the development of an integrated approach to care for these disorders in both the public and private sectors. While some state departments of mental health have developed integrated systems of care for public sector patients, no department appears to have developed such a system for both public and private clients, and there appears to be no published journal report of a model to induce cooperation by all stakeholders. This article outlines a two step approach by the Massachusetts Department of Mental Health to foster stakeholder cooperation in designing an integrated system of care for both public and private clients with co-occurring disorders. PMID- 10695246 TI - The Nexum: a modest proposal for self-guardianship by contract: a system of advance directives and surrogate committees-at-large for the intermittently mentally ill. PMID- 10695247 TI - The liberal neutrality of living and dying: bioethics, constitutional law, and political theory in the American right-to-die debate. PMID- 10695248 TI - HIV-infected surgical personnel under the ADA: do they pose a direct threat or are reasonable accommodations possible? PMID- 10695249 TI - Cedar Rapids Community School District v. Garret F.: the court ruled. Congress, now it's your turn. PMID- 10695250 TI - A national epidemic, a national conversation, a national law: in support of unique identifier reporting for HIV surveillance. PMID- 10695251 TI - Federal gun storage legislation: will this keep guns out of the hands of our children? PMID- 10695252 TI - The Adoption and Safe Families Act of 1997: changing child welfare policy without addressing parental substance abuse. PMID- 10695253 TI - Supervisors beware: the Family and Medical Leave Act may be hazardous to your health. PMID- 10695254 TI - Elevated risk of tuberculosis by occupation with special reference to health care workers. AB - We conducted a study to evaluate tuberculosis (TB) risk in Japan by work performed, either paid or unpaid. We collated information on sex, age, employment category, occupation, and family history from 1120 registration cards of new TB cases at two wards in Nagoya City over seven years (1989-1995). We used census data and data from the Survey of Physicians, Dentists and Pharmacists conducted in 1990 to estimate the population at risk by employment category and occupation. Elevated TB incidence rates were observed for female nurses (SIR: 3.81; 95% CI: 1.97-6.65), clinical laboratory technicians (SIR: 25.00; 6.81-63.99), and males without a paid job (SIR: 1.35; 1.20-1.53). A work environment conducive to transmission may have increased the TB risk in female nurses and clinical laboratory technicians. Male jobless people and institutionalized elderly residents may have enhanced the TB risk for males without a paid job. PMID- 10695255 TI - Yields and daily consumption of cigarettes in Japan in 1969-1996. AB - Cigarette modification trends and the relationship between nicotine yields and consumption in Japan were examined over the 27 years between 1969-1996. Data on cigarette use were obtained from reports published by the government and tobacco manufacturers. Over the study period, there has been a coherent pattern of cigarette modification in Japan. The sales-weighted average yields have declined from 20.7 mg tar and 1.64 mg nicotine/cigarette in 1969 to 8.7 mg tar and 0.72 mg nicotine/cigarette in 1996. On the other hand, the average daily consumption per smoker has continuously increased over the same period. Average nicotine yields and daily cigarette consumption have significant negative correlations among both males and females. This relationship was observed even after controlling for the price changes of cigarettes over time. It is indicated that smokers have compensated for reduced nicotine yields by increasing daily consumption. This may have offset potential benefits of the continuous decline in tar and nicotine yields to smokers' health. PMID- 10695256 TI - Smoking behaviors and attitudes among school teachers in Mie, Japan. AB - We conducted a questionnaire survey of public kindergarten, elementary and high school teachers in Mie Prefecture, concerning smoking habits and attitudes from November 1995 to February 1996. A self-reporting questionnaire was sent to approximately 16,000 teachers and school employees. The questionnaires were collected in a way which took into consideration the privacy of the respondents. A total of 13,998 questionnaires were returned. The percentages of smokers among the teachers were 44.7% for males and 3.1% for females, percentages which are lower than those for the general Japanese population. Almost all of the men and women agreed that anti-smoking education is needed. Most of those who did not feel anti-smoking education was needed were smokers themselves. Seventy percent of both men and women responded that anti-smoking education was a teachers' duty, however, only thirty-six percent of the male and twenty-one percent of the female teachers had actual experience at such education. Finally, almost all teachers wish wish that schools were totally smoke-free or had a partial ban on smoking and believe that school anti-smoking policies in Japan should be introduced. PMID- 10695257 TI - Effects on mortality of getting the basic health examination under the Health Services for the Elderly Act and modification of the effects by health status among elderly persons in a rural community. AB - This longitudinal study examines the effect on mortality of regular use of the basic health examination under the Health Services for the Elderly Act and heterogeneity of the effect according to levels of physical and mental health status among community dwelling elderly persons. Of persons aged 65 and older who lived in Otsuki town, Kochi prefecture, and completed a questionnaire survey about health in February 1991, 1,470 survivors on the anniversary of the baseline survey were followed by the end of March 1996. Regularity of getting the examination was determined by the history of use of the examination in 1990 and 1991. Mortality reduction associated with annual use of the examination was observed in both the 65-74 and the 75 and older age groups and the benefit got smaller with advancing in age. In the 75 and older age group, the benefit from annual use of the examination was restricted to persons having no impairment in physical activities of daily living and those having favorable mental health. Biennial use of the examination was associated with the same amount of mortality reduction as annual use among persons having chronic conditions under treatment in the 65-74 age group. Regular use of the basic health examination at old ages is effective and the effectiveness varies by age range and level of functional health status. PMID- 10695258 TI - Prevalence of idiopathic hypoparathyroidism and pseudohypoparathyroidism in Japan. AB - A nationwide epidemiologic survey of idiopathic hypoparathyroidism and pseudohypoparathyroidism was conducted in 1998 to clarify the prevalence of the two disorders in Japan. From a total of 14,100 departments of pediatrics, internal medicine, neurology, and endocrinology in whole Japan, 2952 (20.9%) study departments were selected at random. Of these departments receiving the first questionnaire, 1855 (62.8%) responded. From these departments 390 patients with idiopathic hypoparathyroidism and 203 with pseudohypoparathyroidism who visited the hospitals in 1997 were reported. The total numbers of patients were estimated to be 900 (690-1100) for idiopathic hypoparathyroidism and 430 (330 520) for pseudohypoparathyroidism (95% confidence intervals in parentheses). Using these data, the period prevalence of the diseases were 7.2 (5.5-8.8) per million population in idiopathic hypoparathyroidism, and 3.4 (2.6-4.2) in pseudohypoparathyroidism (95% confidence intervals in parentheses). PMID- 10695259 TI - Relationship between the blood coagulation-fibrinolysis system and the subclinical indicators of arteriosclerosis in a healthy male population. AB - A cross-sectional observation was performed to assess the relationship between the coagulation-fibrinolysis system and the subclinical indicators of arteriosclerosis in a healthy male population. Subjects were 445 workers (18.9 49.4, Av. 36.2 yrs) in viscose rayon manufacturing factories in Japan. Coagulation-fibrinolysis parameters determined were D-dimer(DD), thrombin antithrombin III complex (TAT), tissue plasminogen activator (TPA), and plasminogen activator inhibitor 1 (PAI1). The following indicators of arteriosclerosis were examined; systolic and diastolic blood pressure (SBP, DBP), stiffness parameter of the carotid artery using ultrasound (beta), pulse wave velocity of the aorta (PWV), and a number of lacunar infarctions from brain MRI. After age-stratification(-29, 30-39, 40+ yrs), the subjects were classified into quartiles by coagulation-fibrinolysis parameters. The mean values of SBP and DBP and beta and PWV, the prevalence of brain infarctions were compared across these quartiles by means of analysis of variance, chi-square test, respectively. Multivariate analysis was also employed to adjust other risk factors. In conclusion, SBP and DBP and beta, PWV were elevated by increase of PAI1, TAT, respectively, in the 40+ years group even after adjustment for other possible risk factors. DD had no relation to any of the indicators of arteriosclerosis. None of the coagulation-fibrinolysis parameters had any relation to brain infarctions. PMID- 10695260 TI - Exposure to Japanese cedar pollen in early life and subsequent sensitization to Japanese cedar pollen. AB - The effect of exposure to Japanese cedar pollen (JCP) in early life on subsequent sensitization to it was evaluated. Specific IgE antibody to JCP was examined in 440-504 school children in a rural town each year during 1995-98. The amount of dispersed pollen measured by a Durham sampler widely ranged from 165 to 5941 grains/cm2/year during this period. The amount had been measured during the period of 1982-91 in which these children were born, and it also widely ranged from 148 to 8566 grains/cm2/year. Children born during November to January, who were exposed to JCP within 6 months of age, increased at the risk of sensitization to JCP, especially severe sensitization, relative to those born in the other months. Age-adjusted prevalence rate ratio (RR) of having a JCP-IgE > or = 15 U/ml (control; < 0.35 U/ml) for children born in December to February relative to children born in the other months was 1.74 (95% confidence interval; 1.06-2.87, examined in 1998), and for those born in November to January was 1.57 (95% CI; 1.00-2.46, examined in 1997). The risk of sensitization to JCP was low for those born in May to July (RR = 0.42, 95% CI; 0.19-0.93, examined in 1998). There was also a strong correlation between the amount of the dispersed pollen during the period of 2-6 months after birth and the prevalence of sensitization to JCP. PMID- 10695261 TI - Long-term prognosis of patients with ulcerative colitis in Japan. AB - Even though it is important in improving the quality of life to evaluate the long term prognosis of patients with ulcerative colitis in comparison with the general population, it is unknown in detail. One hundred and seventeen cases followed-up for 10 years or more were evaluated to define the long-term prognosis of ulcerative colitis by the person-years method. The estimated number of death (E) was 14.5, and the observed number of death (O) was 20. The O/E ratio was 1.38 and confidence interval was 0.84-2.13, showing no significant difference between E and O. Evaluation of change in the O/E ratio at 5-year intervals revealed a decrease in both the males and females, with a significant difference observed between the ratios in 1960 and 1965 (each p < 0.05). Generally, the death rate was significantly higher in the patients with ulcerative clitis than in the general population between 1953 and 1965 but did not significantly differ thereafter. On the other hand, the E from malignant tumors was 3.94, and the O was 4; the O/E ratio was 1.02 and 95% confidence interval was 0.27-2.60, showing no significant difference between E and O. PMID- 10695262 TI - Randomized controlled trial of exercise training for older people (Sendai Silver Center Trial; SSCT): study design and primary outcome. AB - Physical exercise is expected to improve and maintain physical function in older people, thus promoting health and preventing or postponing the onset of disability in later life. The Sendai Silver Center Trial (SSCT) was a randomized controlled trial designed to evaluate the efficacy of exercise training among healthy free-living older people. Sixty-five eligible participants, aged from 60 to 81 years, were randomly allocated to an exercise group or a control group. The subjects in the exercise group were asked to attend training classes at the Sendai Silver Center, a municipal health and welfare facility in the center of Sendai City, at least twice a week for 25 weeks. Each training class, lasting two hours, started with a warm-up session, followed by an endurance session with a bicycle ergometer, and a resistance exercise training session using rubber films, and ended with a cool-down session. The subjects in the control group were asked to attend recreational classes at the Center twice a month. There were no drop outs or accidents during the intervention. Comparison of maximum oxygen consumption (VO2max) before and after the 25-week intervention revealed a significant increase in the exercise group (2.1 ml/kg/min) but no significant change in the control group. Our result is equivalent to the participants becoming younger in aerobic capacity by five years after six months of exercise training. PMID- 10695263 TI - Delays and continuation of hospital visits among HIV-infected persons and AIDS cases in Japan. AB - This study attempts to clarify the distribution patterns of delay between HIV transmission and the first hospital visit among HIV-infected persons and AIDS cases in Japan except those infected through blood products. Such hospital visit patterns were analyzed, and the rates of reporting for HIV/AIDS surveillance among diagnosed HIV-infected persons and AIDS cases in hospitals were shown. From 1991 to 1997, a survey and subsequent follow-up were conducted among HIV-infected persons and AIDS cases diagnosed at 74 hospitals in Tokyo. The numbers of HIV infected persons and AIDS cases were 590 and 208, respectively. The percentage of patients whose estimated date of HIV transmission was obtained ranged 23-41% among Japanese and non-Japanese HIV-infected persons and AIDS cases. Among these patients, 28% to 86% showed a 3-year delay between HIV transmission and their first hospital visit. The rate of HIV-infected persons who continued to visit hospitals within 1 year after their first visit was 77% for Japanese and 45% for non-Japanese; among those after 1 year or more following their first hospital visit the rate was more than 80% among Japanese and over 70% among non-Japanese. The rate of reporting to HIV/AIDS surveillance among diagnosed HIV-infected persons and AIDS cases was 90% or more after 1994 in Japan. The delay between HIV transmission and the first hospital visit was suggested to be very long. Not a few patients stopped visiting hospitals after only a short time. Most diagnosed HIV-infected persons and AIDS cases were reported to the surveillance system of Japan. PMID- 10695264 TI - Bernadine Healy, MD: President of the American Red Cross. PMID- 10695265 TI - The relation between erythrocyte volume and folate levels is influenced by a common mutation in the methylenetetrahydrofolate reductase (MTHFR) gene (C677T). AB - BACKGROUND: The enzyme 5,10 methylenetetrahydrofolate reductase (MTHFR) plays an important role in folate metabolism and folate-dependent reactions. Homozygosity for a common polymorphism in the MTHFR gene (C677T, Ala to Val) is associated with an increased risk of neural tube defects and hyperhomocysteinemia in individuals with low folate levels. Homozygous carriers of the polymorphism with adequate folate levels, on the other hand, seem to be at lower risk for colorectal cancer. Homozygous carriers of the polymorphism (5-15% of the white population) probably represent a subpopulation with increased folate needs. Hematological sequelae of folate deficiency have been recognized for a long time. However, no data exist concerning the relation between the C677T MTHFR polymorphism, folate levels, and hematological parameters. METHODS: We investigated associations between the C677T MTHFR polymorphism, folate levels, total plasma homocysteine, and hematological parameters in 94 patients with cerebrovascular disease (transient ischemic attack/minor stroke) and in 82 healthy subjects. RESULTS: Homozygous carriers (VV) of the polymorphism with low folate levels showed significantly higher homocysteine levels than mutation negative (AA) and heterozygous (AV) subjects (P = 0.038). Furthermore, VV subjects in the lowest folate quartile exhibited significantly higher mean erythrocyte volumes (MCV) and a tendency towards higher erythrocyte hemoglobin content (MCH) than AA and AV subjects (P = 0.008 and 0.069, respectively). Although MCV was not influenced by folate levels in AA and AV subjects, in VV subjects a significant inverse correlation with folate levels could be demonstrated (P = 0.544 and 0.020, respectively). CONCLUSION: We demonstrate an association between the C677T polymorphism, folate levels, and hematological parameters. The elevation of MCV in homozygous carriers of the polymorphism with low folate levels indicates impaired DNA synthesis and/or methylation in these subjects. Considering our data and the results of previous studies, the polymorphism may have contrary effects on homocysteine metabolism and DNA synthesis/methylation dependent on a subject's folate supply. Although the polymorphism is disadvantageous in homozygous carriers with low folate levels, its presence may be beneficial in individuals with adequate folate supply. PMID- 10695266 TI - Soluble P-selectin as a marker of platelet hyperactivity in patients with chronic obstructive pulmonary disease. AB - A comparative analysis between soluble (s) P-selectin and von Willebrand Factor (vWF) was performed in 40 patients with chronic obstructive pulmonary disease (COPD) and 20 healthy subjects, with the aim of investigating whether the occurrence of elevated levels of sP-selectin may reflect activation of platelets, endothelial cells, or both. Plasma sP-selectin levels were significantly higher in patients compared to controls (P < 0.01). Similarly vWF levels were elevated in patients compared to healthy subjects, although the difference did not reach statistical significance. Lipoprotein (a) [Lp(a)] levels were lower in COPD patients than controls (P < 0.0001). The analysis of the correlation among all the variables demonstrated that plasma sP-selectin did not correlate with vWE. Conversely, plasma sP-selectin levels significantly correlated with either oxygen (rho = -0.41, P < 0.05) or carbon dioxide (rho = 0.47, P < 0.05) tension. An inverse correlation between serum Lp(a) and plasma sP-selectin levels (rho = 0.35, P < 0.05) was also observed. Moreover, increasing levels of sP-selectin in COPD patients significantly correlated with the impairment of blood gas tensions. In conclusion, the results obtained indicate the prominent platelet origin of circulating sP-selectin, suggesting that sP-selectin might be considered a marker of in vivo platelet activation in patients with COPD. PMID- 10695267 TI - Autoantibodies from patients with idiopathic ataxia bind to M-phase phosphoprotein-1 (MPP1). AB - In an attempt to identify unique disease-related autoantibodies, the serum from an ataxia and sensory neuropathy patient was used as a probe to isolate a 2.5-kd cDNA from a HeLa expression library. The nucleotide sequence was 99% identical to MPP1, a cell-cycle-related nuclear protein phosphorylated during mitosis. Expression of the cDNA in an in vitro translation system yielded a recombinant protein that migrated in SDS-PAGE at approximately 97 kd. This protein was immunoprecipitated by the prototype human serum, by an immune guinea pig anti MPP1 serum, but not by normal human serum or preimmune guinea pig serum. Western blot analysis of HeLa cell proteins showed that the prototype human serum and immune guinea pig antiserum recognized an approximately 225-kd protein, suggesting that the isolated clone contained a partial cDNA. By indirect immunofluorescence, the affinity-purified antibody and a guinea pig antiserum reacted with nuclei of interphase HEp-2 cells and the cytoplasm of certain neuronal cells. Sera from 10 of 25 unselected patients with ataxia, 1 of 30 patients with peripheral neuropathy, 1 of 50 multiple sclerosis patients, 0 of 20 amyotrophic lateral sclerosis, 0 of 10 children with postviral ataxia, 0 of 10 systemic lupus erythematosus patients, 0 of 3 patients with hereditary cerebellar ataxia, 0 of 8 with ataxia telangiectasia, and 0 of 30 age- and gender-matched controls immunoprecipitated the recombinant MPP1 protein. None of the patients with anti-MPP1 antibodies had evidence of malignancy. This is the first report of MPP1 as a target autoantigen in patients with idiopathic ataxia. PMID- 10695268 TI - Effects of intra-VMN mianserin and IL-1ra on meal number in anorectic tumor bearing rats. AB - BACKGROUND: Tumor growth in animals and humans is associated with the onset of anorexia and reduced food intake. We previously demonstrated that the ventromedial nucleus of hypothalamus (VMN) plays a contributory role in mediating cancer anorexia. Because serotonin and interleukin-1 (IL-1) are putative mediators of cancer anorexia, we hypothesized that their influence on food intake during tumor growth might occur via their action within the VMN. METHODS: To test this hypothesis, 12 Fischer rats injected subcutaneously with 10(6) viable MCA sarcoma cells (TB rats) and their nontumor-bearing controls (NTB, n = 13) were studied. When anorexia developed, TB and NTB rats received bilateral intra-VMN microinjections of the serotonin antagonist mianserin (200 nmol) or the IL-1 receptor antagonist (IL-1ra, 25 ng). Food intake and its determinants of meal number and size were continuously recorded via a computerized device. RESULTS: In NTB rats, intra-VMN mianserin did not affect food intake, whereas after IL-1ra or vehicle a momentary decrease in food intake due to a predominant reduction of meal size occurred. In TB rats, intra-VMN mianserin or IL-1ra selectively increased meal number, leading to improved food intake. CONCLUSIONS: Data suggest that intra-VMN serotonin and IL-1 are involved in influencing cancer related anorexia. PMID- 10695269 TI - Mechanism for the prevention of cholestasis involving cytochrome P4503A overexpression. AB - BACKGROUND: To clarify the preventive effect of taurohyodeoxycholic acid on liver cholestasis induced by toxic bile acids in rats, we evaluated whether modulation of cytochrome P4503A-linked oxidases is involved in the hepatic bile acid retention and secretion mechanism. We investigated whether the safe or the toxic taurochenodeoxycholic acid, administered singly or together, affects cytochrome P450-catalyzed drug metabolism or biliary parameters. We also considered whether the inhibition of the P-glycoprotein export pump by vinblastine might be related to cytochrome P4503A overexpression. METHODS: Hydroxylation of testosterone and N demethylation of aminopyrine were studied in subcellular rat liver preparations after intravenous infusion of hepatoprotective and toxic bile acids administered singly or together. Bile flow, calcium secretion, biliary enzymes activity, and secretion rates of the endogenous and administrated bile acids were determined. CYP3A-dependent monooxygenases were also measured in the same coinfusion model in the presence of vinblastine. RESULTS: Although wide modulation of the activities of different P450 subfamily of isoenzymes was seen, P4503A-associated monooxygenases showed similar patterns in the various situations, i.e., induction by taurohyodeoxycholic acid, reduction by taurochenodeoxycholic acid, and protection (intermediate induction) in the coinfusion experiments. This correlates well with biliary parameters demonstrating the hepatoprotective ability of taurohyodeoxycholic acid. Coadministration of bile acids and vinblastine significantly modifies CYP3A-linked activities. CONCLUSIONS: Bile acid structure seems to be linked with hepatotoxicity/hepatoprotection and P4503A modulation. Taurohyodeoxycholic acid could be therapeutic in cholestatic liver disease by inducing P4503A; we can hypothesize that an associated P-glycoprotein expression might facilitate biliary excretion of toxic taurochenodeoxycholic acid accumulated in the liver during cholestasis. PMID- 10695270 TI - Immediate early gene 2 of human cytomegalovirus increases interleukin 2 receptor alpha gene expression. AB - Previously, we demonstrated that the cytomegalovirus (CMV) immediate early 2 (IE2) gene product upregulates interleukin-2 receptor-alpha (IL-2R alpha) gene expression. To further define this effect, we used a series of IL-2R alpha promoter truncations to identify that the primary site of CMV IE2 activity was in the region between -281 and -273 of the IL-2R alpha promoter, an area without known transcription factor activity. Deletion of known transcription factor enhancer elements resulted in a similar decrease in IE2-driven promoter activation. This unique sequence from the IL-2R alpha promoter was shown to drive a minimal promoter in the presence of IE2. These studies identify a new transcription factor binding site in the IL-2R alpha promoter, which may be specifically responsive to the CMV IE2 gene product. These studies also suggest that one mechanism whereby CMV infection may result in organ transplant rejection is through increased IL-2R alpha expression. PMID- 10695271 TI - Human alveolar epithelial cells type II are capable of regulating T-cell activity. AB - Alveolar epithelial cells type II (AEC-II) express MHC class II on their surface, an important prerequisite for antigen presentation. However, accessory signals are required for an efficient T-cell activation. We therefore isolated AEC-II from tumor-free sections of human lungs obtained by lobectomy/pneumectomy and purified the cells by magnetic-activated cell sorting. Furthermore, we tested the expression of CD54, CD58, CD80, and CD86 on AEC-II and evaluated their accessory function (AF) in cell culture using a coculture of interleukin-2 (IL-2), releasing Jurkat cells and AEC-II. An increased AF is documented by an elevated IL-2 release and expressed as accessory index (AI). In 33 experiments the AF of AEC-II proved to be highly variable. AI ranged between 0.3 and 17.1 with a median of 1.4 (0.3-17.1). Forty-four percent (4-77) of the AEC-II expressed HLA-DR, 44% (12-89%) of the cells expressed CD58, and CD54 was expressed by 55% (16-89%). AEC II also expressed CD80 and CD86 (38% [0-77%] and 40% [4-68%], respectively). Interestingly, AEC-II released high levels of TGF beta (1730 pg/mL [771-5876]) and the accessory index could be increased (approximately 2-fold) by the addition of neutralizing anti-TGF beta antibodies or radiation. Thus, type II alveolar cells express costimulatory molecules and are able to deliver costimulatory signals for T cells, providing evidence that AEC-II are able to act as antigen presenting and immunoregulatory cells of the lung. Additionally, the accessory function of AEC-II is under the control of endogenously released TGF beta. PMID- 10695272 TI - The public's health at the beginning of the century--challenges and opportunities. PMID- 10695273 TI - Public health in the commonwealth 100 years ago. 1940. PMID- 10695274 TI - Kentucky State Board of Health biennial report. 1906-07. PMID- 10695275 TI - Community assessment using the key informant method: a snapshot of some rural communities from the perspective of community leaders. PMID- 10695276 TI - Problems engendered by illegible medical records needs to be addressed. PMID- 10695277 TI - The leadership role of the medical family in this century. PMID- 10695278 TI - HLA diversity and infections. PMID- 10695279 TI - Rural-urban differences in food and nutrient intake of Pakistani children. AB - BACKGROUND: Global increase in urbanisation accompanied by increase in complexity of nutritional problems is a cause of concern for most nations. OBJECTIVE: The aim of this study was to assess the differences in frequency of food consumption and nutrient intake of urban and rural Pakistani children. SETTING: Forty rural, 59 middle income urban and 81 affluent urban children belonging to the province of Punjab. Children were recruited through schools in Lahore and Rayonpura, Kala Shah-Kaku (Sheikhupura district). METHOD: Three-day estimated diet records were kept by a total of 180, 10-12 year old school-children. The nutrient intake was calculated by the nutritional analysis package COMP-EAT and analysed on SPSS. The results of the three groups were compared. RESULTS: Consumption of chapati, eggs, yoghurt and some of the traditional vegetarian snacks was very similar in all the three groups. Consumption of lentils, cooked vegetables, paratha (fried Asian bread) and tea decreased and that of milk, meat curry, chicken curry, chocolates, cakes, ice-cream, fruit and raw vegetables increased with urbanization. As compared with the rural children, the urban children had a higher mean daily intake of calories, sugar, protein, total fats, cholesterol, calcium, sodium, potassium, niacin, vitamin B12, folic acid, antioxidant vitamins A, C and E and lower intake of total carbohydrates, fibre and starch. CONCLUSION: It is concluded that although the macro-nutrient consumption pattern of rural children appears to be heart healthy lower consumption of protective micro-nutrients by them may put them at risk. In view of rapid urbanisation and its multidimensional impacts on the health of the populations living in the urban areas of the developing world, these dietary trends provide baseline information for health professionals. PMID- 10695280 TI - The clinical pattern of HER-2/neu oncogene overexpressing breast cancer in Pakistani patients at initial presentation: an analysis of HER-2/neu positive versus negative disease--a preliminary report. AB - BACKGROUND: HER 2/new oncogene is an important prognostic marker in Breast Cancer and has implications in therapy planning. OBJECTIVE: To describe the clinical features of HER 2/new positive and negative Breast Cancer in the Pakistani patient population and note clinical differences between the two groups if any. DESIGN: A retrospective analysis of Breast Cancer cases at the Aga Khan University, Hospital. METHOD: Immunochemical staining on formation fixed paraffin embedded tissue using oxidase antiperoxidase method. A total of 152 Breast cancer tissue samples were tested for HER-2/neu gene presence. Of these 43 (39%) samples tested positive and 109 (61%) tested negative. A comparison of the two groups revealed that only a few factors tested for either significance or borderline statistical significance between the two groups. These factors included the estrogen receptor status and the number of lymph nodes involved in the axilla. The progesterone receptor status was of borderline significance. CONCLUSION: Given the large number of factors tested it appears that there is no consistent defining feature which helps to separate HER-2/neu positive versus HER-2/neu negative cases with Breast Cancer. PMID- 10695281 TI - Self-reported feeding advice by physicians for common childhood illnesses. AB - BACKGROUND: A nutritious diet is important for recovery during illnesses. Dietary advice by physicians and consumption of food by the patients are often based upon their hot and cold concepts and beliefs about various foods rather than on scientific basis. OBJECTIVES: To look at the food-advising behaviour of physicians during illnesses and to know the maternal concepts about various foods being hot or cold. METHODS: A questionnaire was served to the physicians participating in a continuous medical education session held at the Aga Khan University and Hospital, asking them to write the foods they advise or restrict during different illnesses such as fever, jaundice etc. Mothers of sick children suffering from diarrhea and other illnesses were also interviewed to know their concepts about various foods. RESULTS: Six (10%) out of sixty physicians believed in hot and cold concepts of the food. A variety of common foods were either restricted or strongly recommended by 10% to 50% of these physicians without any scientific basis, 23% physicians restricted fatty foods in jaundice, 17% physicians restricted in cough and cold. Although the interviewed mothers believed in hot-cold concepts of food but 55-63% of them were not sure what is meant by hot or cold food. In spite of that 70-80% of these mothers classified chicken, meat, egg, brinjal, masoor and mangoes are hot foods and rice, yogurt, moong, banana and orange as cold foods. CONCLUSION: Hot-Cold concept of food exists not only in mothers but also in physicians. Proper education regarding food intake is mandatory for both mothers and physicians to ascertain adequate intake of calories during sickness. PMID- 10695282 TI - Immunization coverage in three districts of North West Frontier Province (NWFP). AB - AIM: This community-based study was conducted to assess the progress of Expanded Programme on Immunization (EPI) in North West Frontier Province (NWFP) of Pakistan. METHOD: In this cross sectional survey, 120 randomly selected clusters in 03 districts of NWFP were included, 2673 children in the age group 12-35 months of 2583 randomly selected families were visited. RESULTS: The results showed that 65% of children were fully immunized, but out of them only half could be verified by immunization, need of 2nd and 3rd dose and no faith in immunization were the major causes of failure of immunization programme. Moreover, mother too busy, absence of vaccinator and inconvenient place of immunization were the obstacles pointed out by the parents. CONCLUSION: This study suggests the areas where improvement can be made to achieve the real target of immunization coverage. It is concluded that despite of more than 20 years of efforts by EPI, the ultimate objectives have not been achieved. PMID- 10695283 TI - Gastric carcinoma with lymphoid stroma: association with Epstein virus genome demonstrated by PCR. PMID- 10695284 TI - Endoscopic placement of esophageal prostheses for inoperable carcinoma of esophagus. PMID- 10695285 TI - Gender issue neglected aspect of health promotion in Pakistan. AB - OBJECTIVE: To identify neglected aspects of primary health care activities and propose effective strategies for better health promotion in Pakistan. METHOD: An observation study was carried out from March-July, 1998 in a low-medium income group in Karachi West in a primary health care set-up. RESULTS: Seventy-three percent of female clients were influenced by the advice of male members of the family, 22% followed the directions of elderly female member, 5% availed health services on their own will. CONCLUSION: The Pakistani female community is influenced strongly by the male members of the family in almost all health related activities. PMID- 10695286 TI - Prognostic indicators of childhood acute viral encephalitis. AB - OBJECTIVE: To devise a set of clinical signs and laboratory parameters that would help clinicians assess prognosis in patients and plan appropriate management. METHODS: Medical records of 147 paediatric cases (with a discharge diagnosis of acute viral encephalitis) admitted over a ten year period from 1987 to 1997 were reviewed and relevant information collected on a data extraction form. RESULTS: Of 147 patients, 24 (16.3%) died and 48 (32.7%) were left with severe neurological deficits. A GCS (Glasgow Coma Scale) score between 6-10 had an association with poor outcome (OR = 2.62, Chi-square = 5.57, p-value = 0.018) and that a GCS score of > or = 5 was even more strongly suggestive of poor outcome (OR = 5.49, Chi-square = 12.08, p-value = 0.0005). A history of having seizures, for more than 3 days, also showed a strong association with poor outcome (OR = 3.66, Chi-square = 5.46, p-value = 0.019). CONCLUSION: Patients with an increased risk of death and severe disability can be identified using a few guidelines. Of these, a history of seizures of > 3 days and/or impaired consciousness (GCS < or = 10), at the time of presentation to the hospital, constitute high risk. These cases must be identified promptly and aggressive therapy initiated in order to improve long term outcome. PMID- 10695287 TI - Characterization of Streptococcus pneumoniae strains isolated from systemic infections in children. AB - Twenty-three cases of systemic pneumococcal infection diagnosed from October 1988 to September 1998 were analyzed retrospectively in order to characterize the epidemiology of systemic pneumococcal infections. The clinical diagnosis of those cases were 8 pneumonia, 8 meningitis, 3 septicemia, 3 septic arthritis, and 1 peritonitis. The patients ranged in age from 6 months to 21 years old (mean +/- SD = 3 years, 6 months +/- 5 years, 2 months), and 61% of the patients were younger than 24 months. Resistance to penicillin G (PCG) was detected in 57% of all cases. Resistance to cefotaxime (CTX), imipenem (IPM), erythromycin (EM), and clindamycin (CLDM) was 33%, 9%, 70%, and 65%, respectively. Of the 13 isolates resistant to PCG, 2 were resistant to IPM, 11 to EM and 11 to CLDM. Serotyping was performed on 17 isolates. The identified serotypes were 19 (6 isolates), 6 (5 isolates), 23 (3 isolates), 14 (2 isolates), and 5 (1 isolate). Eleven isolates resistant to PCG were limited to serotypes 6, 19, or 23. One patient had a recurrent episode of bacteremic pneumonia 7 months after the first episode. Streptococcus pneumoniae isolates from both episodes were compared by serotyping and pulsed-field gel electrophoresis with restriction digestion, and were confirmed as the same strain. PMID- 10695288 TI - [A trial of amantadine for the treatment of influenza A infection in children]. AB - We administered amantadine to 52 children for therapy and 3 children for prophylaxis of influenza A infection from January to March 1998. Among the 23 children in the therapy group with proven influenza A/H3N2 infection, 15 patients (60%) allayed fever within 3 days but 9 (40%) had persistent fever for more than 4 days after administration of amantadine. Seven of these nine patients (40%) received amantadine within 2 days after the onset of fever. Two of these nine had secondary bacterial infections. One child had insomnia as side effect. We concluded that administration of amantadine for therapy is safe and effective, if given to patients without secondary bacterial infections in the first 48 hours of the illness. PMID- 10695289 TI - [Evaluation of the rapid detection test for influenza A and B viruses using neuraminidase activity]. AB - The ZstatFlu test (ZymeTx, USA) is a rapid detection kit for influenza A and B viruses. This test is based upon the reaction between viral neuraminidase from influenza viruses and a chromogenic substrate. The clinical performance of the ZstatFlu test was determined by comparison with viral isolation in cell culture. A total of 176 respiratory specimens from 172 pediatric patients with influenza like illnesses during the 1998/99 season were tested. Influenza viruses were recovered from 97 specimens (type A: 6, type B: 91) in cell culture. ZstatFlu demonstrated 67.4% sensitivity (29/43) and 62.7% specificity (37/59) for throat swabs. Of the 22 ZstatFlu-positive, culture-negative throat swabs tested by RT PCR, 18 were positive by RT-PCR. ZstatFlu showed 48.1% sensitivity (26/54) and 90.0% specificity (18/20) for nasopharyngeal aspirates. Of the two ZstatFlu positive, culture-negative nasopharyngeal aspirates tested by HI titer of paired sera, one showed a 4-fold increase of HI titer. Nasopharyngeal aspirates therefore showed lower sensitivity than throat swabs at this test, different from EIA test kits such as Directigen FluA or FLU OIA. Overall, only 5 specimens were false positive by the ZstatFlu test. Therefore, this test demonstrated high specificity and positive predictive value. In conclusion, the ZstatFlu test is useful for the rapid detection of influenza A and B viruses to identify patients who need antiviral treatment. PMID- 10695290 TI - [The efficacy of influenza vaccine among geriatric inpatients]. AB - To investigate the efficacy of influenza vaccine in the elderly, hemagglutination inhibition (HI) antibody titer for the three types of influenza viruses were measured and the influenza infection rate was determined serologically in geriatric inpatients. Influenza vaccination was done for inpatients. For patients who had influenza vaccination in the year prior to the study, influenza vaccine was administered once or twice, and the number of injections were determined randomly. Influenza vaccine was injected twice to those had not received influenza vaccine in the previous year. Serum samples were collected from 166 vaccinated and 104 unvaccinated patients before and after 1996/1997 influenza season. In the vaccinees who had been vaccinated the previous year, 56 patients were injected once and 58 patient were injected twice. Fifty-two patients had not been vaccinated the previous year. Serologically diagnosed influenza infection rate in the 104 unvaccinated patients was 16.3% for influenza A/H3N2 and 8.7% for influenza B. The infection rate was 3.0% for influenza A/H3N2 and 0.6% for influenza B in the 166 vaccinated patients. The infection rates were significantly lower in the vaccinees than in the unvaccinated patients (p < 0.001 with A/H3N2 and p < 0.01 with B). There was no significant difference in the infection rate among the three vaccinated groups. These results suggest that the influenza vaccination had significant protective efficacy for influenza infection in the elderly. Prior vaccination did not diminish the efficacy of the influenza vaccine. The efficacy of a single influenza vaccine injection was equivalent to that of two injection. PMID- 10695291 TI - [Extended-spectrum beta-lactamase (ESBL) produced by Escherichia coli and Klebsiella pneumoniae isolated from Teikyo University Hospital--the second report]. AB - We studied the high-level resistant to cefotaxime (CTX, MIC > or = 512 micrograms/ml) clinical isolates of Escherichia coli and Klebsiella pneumoniae in Teikyo University Hospital. The CTX-resistance could be transferred to E. coli K 12 chi 1037 or ML4903 strains from 30 of the 33 isolates by conjugation at a frequency of 10(-4). When the hydrolysis rate of benzylpenicillin was 100%, the beta-lactamases which were extracted from the transconjugants hydrolyzed CTX, CAZ and AZT at the rate of 38-95%, 0-8.6% and 0-56%, respectively. These results demonstrate that these enzymes should be categorized into ESBL. The nucleotide sequence of CTX-resistant gene was identified to that of the CTX-M2 gene which was first described in Argentina. It was found to have 99.9% homology to Toho-1 gene in Japan and 99.6% homology to CMY-2 gene. Using a PCR methods for the detection of one of ESBL gene such as CTX-M2, Toho-1 or CMY-2, the DNA was amplified from all strains (11 isolates of E. coli and 21 isolates of K. pneumoniae). PMID- 10695292 TI - [Antibody induction and frequency of adverse reactions to influenza vaccines in the elderly]. AB - A total of 1,223 elderly people in nursing homes in Niigata Prefecture, Japan, were immunized with one or two doses of commercial trivalent split vaccine formulation, against strains including A/HN, A/H3N2 and B for three seasons (1996 1999). The frequencies of adverse reactions and antibody induction were assessed. Frequent side effects of vaccination were local reactions such as redness and tenderness at the site of injection, but there were no serious reactions, suggesting that the vaccine was quite safe for the elderly. Furthermore, antibody induction by immunization was relatively high and independent of the degree of activities of daily living (ADL). Annual repeated influenza vaccination did not diminish protection against influenza. However, antibody induction against antigens was insufficient in the 1997/1998 season, and further improvement in the combination of quantities of the four included antigens may by required. A booster dose after the first dose did not enhance immune responses in the nursing staff, and the one dose method appeared to be indicated for the elderly. PMID- 10695293 TI - [Serovar distributions of cervical Chlamydia trachomatis isolated in Nagano Prefecture]. AB - Between 1992 and 1998, serotyping of 82 Chlamydia trachomatis cervical isolates were examined by micro-IF method in Nagano Prefecture. Of these, 17 isolates (20.7%) were serovar E, 15 (18.3%) were serovar F, 14 (17.1%) were serovar D and 10 (12.2%) were serovar G, and the isolates typed these serovars were found to be 68.3% whole. Furthermore, serovars B (7.3%) and K (9.8%) were comparatively frequently found. From 1992 to 1994, the number of isolates of B-complex, intermediate and C-complex were 18, 17 and 6, respectively. On the other hand, from 1995 to 1998, the number of the isolates were 19, 8 and 14, respectively. The distribution of serovars of C. trachomatis tended to fluctuate from serovars D and E to F and G as the patients grew older. Moreover, serovar E isolates were only detected from patients less than 40 years old. Although most of the serovar I isolates were detected from the patients with vaginal discharge, the other isolates did not clearly indicate the relationship of serovars and clinical symptoms. PMID- 10695294 TI - [Clinical analysis of community-acquired pneumonia requiring hospitalization in a community hospital--comparison of elderly and non-elderly patients]. AB - A comparative study of 890 patients with community-acquired pneumonia requiring hospitalization in a community hospital was performed. The patients were divided into an elderly patient group and a non-elderly patient group. The elderly patients with community-acquired pneumonia exhibited frequent atypical symptoms such as dyspnea, consciousness disturbance and complication of shock, and also were frequently in a poor nutritional condition. The causative microorganism was isolated in 40.8% of the elderly patients and in 44.0% of the non-elderly patients. Polymicrobial agents were detected frequently in the elderly patients. Streptococcus pneumoniae (19.4%), MSSA (16.8%), Klebsiella pneumoniae (15.1%) and Haemophilus influenzae (15.0%) were frequently isolated from the sputum of the elderly patients, while Mycoplasma pneumoniae (25.2%), H. influenzae (15.0%), S. pneumoniae (12.2%) and MSSA (10.2%) were frequently isolated from that of the non elderly patients. Regarding treatment with antibiotics, therapy with a single antibiotic therapy, such as cephem or carbapenem was carried out for the elderly patients, while new quinolone or tetracycline was administered to the non-elderly patients. Although the treatment with antibiotics was adequate according to the guidelines of the American Thoracic Society, the prognosis was poor; i.e.) in the elderly patients an efficacy rate of 74.3% and a mortality rate of 9.5%. In the non-elderly patients, the prognosis was good; i.e.) an efficacy rate of 88.0% and a mortality rate of 1.7%. These results suggest that the most important factors affecting the prognosis were the general condition of elderly patients and delay in an adequate diagnosis and treatment because of atypical clinical findings. PMID- 10695295 TI - [A clinical investigation of infective endocarditis at a community hospital in Japan]. AB - There have been few reports on the clinical features of infective endocarditis (IE) in Japan. We clinically investigates 45 episodes (36 cases) of definite IE that were experienced from January 1985 to March 1997 at a community hospital, Okinawa Chubu Hospital, Okinawa, Japan. Regarding age, prior dental procedure, causative organisms and sites of infection, analyses and comparison were performed on a total of 94 episodes, by adding another 49 episodes of IE that were experienced between 1977 and 1984 at our hospital. The mean age was 47 years and majority of patients in the recent 12 years were older than 50 years of age. Mortality of all 94 episodes was 20%, while that of recent 45 cases was 13%. Eight % of all episodes had history of recent dental treatment but significance of the finding remains unclear. Alpha streptococci were most common (33%) and Staphylococcus aureus was the second most common organism (17%). Eleven % of all episodes were culture-negative and there was a statistically significant difference in the histories of prior antibiotic therapy between culture-negative and culture-positive episodes. Regarding sites of infection, 27% of all episodes involved mitral valves, while 24% involved aortic valves. Prosthetic valves were involved in 12%. Ninety-eight % of the recent episodes had fever, 98% had cardiac murmurs and 27% had characteristic mucocutaneous lesions. Heart failure was the most common complication (27%) and half of the cases with prosthetic valve infection had heart failure. Cerebral embolism was most frequently seen among the major arterial embolic complications. Our results were similar to those which were previously reported from other countries. We should have a high index of suspicion for endocarditis whenever we see patients who present various clinical manifestations and fever of which origin remains unclear. Willingness to obtain blood culture before starting antibiotics is most important. PMID- 10695296 TI - [Gastrointestinal diseases associated with HIV infection]. AB - A clinical studies were carried out on gastrointestinal diseases associated with HIV infection. During the 6 years between January 1993 and December 1998, 71 HIV infected cases visited to Yokohama Municipal Citizen's hospital, and 26 of them developed gastrointestinal complications during the course of their illness. They consisted of 24 males and 2 females, with the mean age of 44.7 years and the medial value of 42.5 years. Of the 26 patients, 21 were Japanese, and the remaining 5 were Southeast Asian. The mean CD4 count was 143/microliter and the medial value was 32/microliter at the time of development of complications. Gastrointestinal complications were esophageal candidiasis in 6 patients, cytomegalovirus (CMV) gastritis and gastric Kaposi's sarcoma in 1 patient each, amebiasis in 8 patients, infectious colitis in 11 patients, and asymptomatic pathogen carriers in 3 patients. Esophageal and gastric complications were common in patients with low count of CD4, and endoscopy was useful for diagnosis. Amebiasis developed even in patients with normal CD4 and was common in males with experience in homosexual contact. It seems that homosexual contact acquire not only HIV infection but also Entamoeba histolytica through sexual contact. Protozoan and acid-fast bacteria were detected at high rate in patients with infectious colitis and asymptomatic pathogen carriers. Besides food-born infections, imported infections were seen in foreign and Japanese patients who had traveled abroad. The gastrointestinal diseases associated with HIV infections for the most part were opportunistic infections or tumors but imported, food born, and sexually transmitted infections were also observed. It seems necessary to take into consideration of varying background of patients in the treatment of gastrointestinal diseases associated with HIV infections. PMID- 10695297 TI - [Geographic distribution of Mycobacterium avium-intracellulare complex serovars isolated from patients in five cities of Japan]. AB - Mycobacterium avium-intracellulare complex (MAC), isolated from patients with MAC infection in five cities in Japan, was classified into M. avium and M. intracellulare by DNA-DNA hybridization assay (DDH). They were also classified into serovars by seroagglutination test and thin-layer chromatography (TLC) detecting serovar-specific antigens. The ratios of M. avium: intracellulare isolated in Sendai, Maebashi, Kobe, Hamamatsu, and Kumamoto, were 100:0, 26:16, 37:12, 37:39, and 17:54, respectively. It was high in the northern city, Sendai, whereas the ratio in the southern city, Kumamoto, was low. The main serovars detected in Sendai and Maebashi were 8 and 4. Various serovars including a new type of M. intracellulare were detected with main serovars 16 and 7 in Kumamoto. M. intracellulare including main serovar 16, and M. avium including main serovar 4, were detected in the same ratio in Hamamatsu. M. avium including main serovars 8 and 4, was detected in considerable high ratio in Kobe. Geographic distribution was observed in both species of MAC and serovars. PMID- 10695298 TI - [Flowcytomeric analysis on neutrophil intracellular enzyme activity in patients on hemodialysis and continuous ambulatory peritoneal dialysis]. AB - To elucidate the mechanism of neutrophil dysfunction in patients with maintenance hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD), intracellular enzyme activity such as oxidative burst, elastase, cathepsin, and collagenase, was investigated. Response of enzyme activity to in vitro addition of TNF-alpha, which is known to have a powerful priming effect on neutrophils, was also evaluated. Peripheral blood from 15 HD and 15 CAPD patients was washed and incubated with Cell Probe, an indicator for intracellular enzyme activity. Mean fluorescent intensity of neutrophils, which represents neutrophil enzyme activity, was measured by flowcytometry. In HD group, unstimulated enzyme activity was similar to that of control, but activity after addition of TNF-alpha was significantly lower than the control. In the group of CAPD, enzyme activity without stimulation was not different from that of control, and in TNF-alpha stimulated neutrophils, only elastase activity was lower than control. Many of the enzyme activities after stimulation were lower in HD than in CAPD. Response to in vitro addition of TNF-alpha was diminished in both dialysis groups, but more prominent in HD neutrophils. Duration of dialysis, serum concentration of beta 2-microglobulin (beta 2-MG) and parathyroid hormone (PTH) was significantly related inversely to intracellular enzyme activity in HD patients. To the contrary, in CAPD group, although beta 2-MG and PTH showed similar negative correlation, duration of dialysis was not related to enzyme activity. These results indicate that neutrophils in patients with maintenance dialysis have diminished intracellular oxidative burst, elastase, and cathepsin activity. Especially, impaired response to TNF-alpha closely related to neutrophil dysfunction in dialysis patients. PMID- 10695299 TI - [Studies on centrifuge-concentration method of pooled samples for HIV screening test by RT-PCR]. PMID- 10695300 TI - Specialists and the current medical "system". PMID- 10695301 TI - The evils in the forest primeval. PMID- 10695302 TI - Family medicine, generalist medicine, specialist medicine and the conduct of the consultive relationship. PMID- 10695303 TI - Primary care: the backbone of prevention. PMID- 10695304 TI - Reflections from the field. PMID- 10695305 TI - A day with Dr. Malaprop (or) on burning your bridges at both ends. PMID- 10695306 TI - Some thoughts on the treatment of psychiatric disorders in the primary care setting. PMID- 10695307 TI - Obstetrics in family practice. PMID- 10695308 TI - Rhode Island's Cardiac Services Registry. PMID- 10695309 TI - Tobacco control report card: Rhode Island, 1999. PMID- 10695310 TI - Improved access for patients or health care fraud? Hospital provision of office space to physicians. PMID- 10695311 TI - Education and science: similarities and differences. PMID- 10695312 TI - Indoor environment and asthma. PMID- 10695314 TI - Environmental air pollution and lung disease in children. AB - Acute exposure to air pollution is associated with increased respiratory symptoms and decreases in lung function in children. Respiratory symptoms in healthy children are usually nonspecific and not severe. Lower respiratory symptoms and extra use of bronchodilators will increase by about one-third with exposure to peak levels of ozone in children with asthma. Similarly, sulphates will increase the use of medication and decrease lung function in asthmatic children. Hospital and outpatient admissions for children with pre-existing asthma may be increased in the range of 20% with acute exposure to ambient ozone peaks and possibly with increased sulphur dioxide (SO2). Short-term nitrogen dioxide (NO2) exposure from indoor and outdoor sources has been associated with nonspecific respiratory symptoms and decreased lung function, again particularly in children with pre existing asthma. No effect on hospital admissions has been documented. Chronic exposure to respirable particles, SO2 and NO2 is associated with up to three-fold increases in nonspecific chronic respiratory symptoms. Exposure to high traffic flow and, in particular, truck traffic and diesel exhaust leads to significant increases in respiratory symptoms and decreases in lung function, while no clear effect on the inception of asthma has been documented. It appears unlikely that long-term exposure to pollutants or irritants is responsible for the increase in asthma and allergy observed in many countries. However, although the effect of air pollution is small in most children, it has a significant influence on the health of children with pre-existing lung disease. Owing to the large number of individuals exposed this results in a considerable burden for the health system. PMID- 10695313 TI - Outdoor air pollution in urban areas and allergic respiratory diseases. AB - Respiratory allergic diseases (rhinitis, rhinosinusitis, bronchial asthma and its equivalents) appear to be increasing in most countries, and subjects living in urban and industrialized areas are more likely to experience respiratory allergic symptoms than those living in rural areas. This increase has been linked, among various factors, to air pollution, which is now an important public health hazard. Laboratory studies confirm the epidemiological evidence that inhalation of some pollutants, either individually or in combination, adversely affect lung function in asthmatics. The most abundant air pollutants in urban areas with high levels of vehicle traffic are respirable particulate matter, nitrogen dioxide and ozone. While nitrogen dioxide does not exert consistent effects on lung function, ozone, respirable particulate matter and allergens impair lung function and lead to increased airway responsiveness and bronchial obstruction in predisposed subjects. However, besides acting as irritants, airborne pollutants can modulate the allergenicity of antigens carried by airborne particles. By attaching to the surface of pollen grains and of plant-derived paucimicronic particles, pollutants can modify the morphology of these antigen-carrying agents and after their allergenic potential. In addition, by inducing airway inflammation, which increases airway epithelial permeability, pollutants overcome the mucosal barrier and so facilitate the allergen-induced inflammatory responses. Moreover, air pollutants such as diesel exhaust emissions are thought to modulate the immune response by increasing immunoglobulin E synthesis, thus facilitating allergic sensitization in atopic subjects and the subsequent development of clinical respiratory symptoms. PMID- 10695315 TI - Environmental lung disease: the role of diet. PMID- 10695316 TI - Tobacco withdrawal symptoms and treatment. PMID- 10695317 TI - Smoking cessation: nicotine replacement, gums and patches. AB - This is an overview of smoking cessation with a clinical approach. The different nicotine replacement products approximately double the long-term success rate with a dose-response effect, but with an equal efficacy of the different administration forms, so there is no long-term effect of a combination of two products, and they have only mild side-effects. The success rate will increase with the level of adjunctive behavioural support. Total smoking abstinence during the first week after quit-day is a very strong predictor of long-term success. Bupropion, an antidepressant drug, seems to be a promising drug in smoking cessation and is at least equally efficacious as nicotine products. All health care workers in pulmonary units have an obligation to deliver up-to-date smoking cessation services to their smoking patients. PMID- 10695318 TI - The psychology of tobacco addiction: why it is difficult to stop smoking. PMID- 10695319 TI - The ELITE system. AB - This report describes the technical limitations that affect the computation of thoraco-abdominal volume displacement and the characteristics that an ideal system should have. The elaboratore di immagini televisive (ELITE) system satisfies many of these characteristics. ELITE system is an optoelectronic device able to track the three-dimensional co-ordinates of a number of reflecting markers placed noninvasively on the skin of the subject. The simultaneous acquisition of kinematic signals with pleural and gastric pressures during a relaxation manoeuvre allows the representation of pressure-volume plots describing the mechanical characteristics of each compartment. The results of studies concerning chest-wall mechanics by applying the ELITE system are described. PMID- 10695320 TI - Negative expiratory pressure method for the detection of expiratory flow limitation. PMID- 10695321 TI - Lung transplantation for emphysema. PMID- 10695322 TI - Pulmonary rehabilitation in patients with severe chronic obstructive pulmonary disease. PMID- 10695323 TI - Terminology and testing of respiratory muscle dysfunction. PMID- 10695324 TI - Respiratory muscles in internal medicine. AB - This review provides evidence that respiratory muscle abnormalities are present in many illnesses of internal medicine and emphasizes that clinicians should look for respiratory muscle weakness in many circumstances, particularly immunological disorders. Controversial results in hormonal diseases, metabolic diseases and abdominal disorders indicate areas for further research. PMID- 10695325 TI - Effects of surgery on the function of the respiratory muscles. AB - The function of the respiratory muscles (RM) is affected positively or negatively by a variety of surgical procedures. Cardiac, thoracic and upper abdominal surgery impair the RM function and lead to postoperative complications such as hypoxia, atelectasis, aspiration and infections. Preoperative assessment of RM function is cardinal to avoid or attenuate these complications. Three types of surgical procedures, lung transplantation, lung volume reduction surgery and surgery for obesity have been shown to improve RM function. A mechanism by which these types of operation have shown beneficial effects on RM function is multifactorial, depending on geometrical factors, from the reduction of hyperinflation and those depending on changes on the control of breathing. Physicians dealing with postoperative care of patients should be aware of the pathophysiological mechanisms that impair or improve respiratory muscle function as a result of a surgery as well as of the therapeutic modalities. PMID- 10695326 TI - Respiratory muscles in intensive care medicine. PMID- 10695327 TI - Structural cells and extracellular matrix in chronic airways inflammation. PMID- 10695328 TI - Are asthma and chronic bronchitis different diseases? Pro. PMID- 10695329 TI - Are asthma and chronic obstructive pulmonary disease different diseases?--Con. PMID- 10695330 TI - [Endoscopic endonasal surgery with image-guidance]. AB - We evaluated the advantages and disadvantages of image-guided endoscopic endonasal surgery for various diseases. Thirty-three patients, including 8 with chronic sinusitis, 14 with paranasal cysts, 1 with paranasal tumor (biopsy), 1 with sellaturcial cyst (Rathke's cleft cyst) and 9 with pituitary tumors were endonasally operated on from September 1998 to May 1999, with an electromagnetic navigation system, The Insta Trak (Visualization Technology Inc. USA). The Insta Trak system is composed of a computer, a metal probe with a nonmetallic suction tube attachment, and a soft-type headset with an electromagnetic sensor. This freehand, armless system compensates well for patient's head movement during surgery, and precludes the need for head fixation. Either straight or curved suction tube (probe) can be used to access almost of all pathological sites in the sinus cavity. Location of the metal probe is displayed on the computer moniter as an intersection point on the axial, coronal and sagital CT images. In all cases, Insta Trak showed the surgeon the appropriate location and direction of each lesion. The Insta Trak also indicated the location of the orbit, optic canal, nasolacrimal duct and/or skull base, thus, preventing intraoperative complications. When the anatomy was distorted by previous surgery and/or when there was uncontrollable bleeding from a severe lesion so that the surgeon had difficulty finding the proper orientation, the usefulness of image-guided surgery was sufficiently recognized. However, the following disadvantages were also pointed out. An additional 15 to 20 minutes is needed for equipment set up and operation, unless the surgeon and the operation room staff are familiar with the machine. The patient's CT image used for navigation relies on data obtained preoperatively, that is to say, it can not reflect morphological changes produced during surgery. Moreover, the surgeon must consider possible errors of the navigational point that may result in the headset during surgery, as well as, errors the machine may originally possess. The image-guided system successfully integrated the most up-to-date computer technology with a surgeon's anatomical knowledge for improved treatment of endoscopic endonasal surgery. However, we also concluded that the system should be used as a surgical supporting device for safer and more adequate procedures. PMID- 10695331 TI - [Management of the acoustic tumor in an only/better hearing ear]. AB - Because profound bilateral hearing impairment is a catastrophic event, the management and care of an individual with an 8th nerve lesion in an only/better hearing ear remains a significant challenge for both patient and physician. Current options regarding the treatment of the acoustic tumor in an only/better hearing ear include: observation, attempted hearing preservation surgery and stereotactically guided radiation therapy. In this article, we present 3 cases of acoustic tumor within the internal auditory canal in an only/better hearing ear diagnosed by gadolinium-enhanced MRI and discuss the recommendations, especially observation, available in the care of these cases. In one patient, hearing disturbance caused by the tumor in a better hearing ear made the patient quite depressive and desperate. One of the most important consideration is for the physician to provide the patient with adequate informed consent regarding the possibility of profound bilateral hearing loss caused by either the natural growth or surgical removal of the tumor in the future, and alternative methods of communication with others such as: hearing aid and lip reading, cochlear implant and brainstem implant. PMID- 10695332 TI - [Ultrasonographic screening for thyroid cancer in the screening]. AB - In our hospital, ultrasonography is performed for patients who come for thyroid cancer screening. Seventy-eight patients with thyroid cancer which was found by such screening were operated on from 1989 to 1998, while 287 patients with thyroid cancer found by other methods were operated on during the same period in our department. The age of the patients at surgery in the screening group was younger than that of the contrast group. In the screening group, 41 (52.6%) patients had small thyroid cancer, a higher rate that in the contrast group. Invasion of surrounding organs by the primary cancer was observed in only one patient (1.3%) in the screening group, a rate of invasion that was statistically lower than that in the contrast group. Twenty-six patients with benign thyroid disease detected in our screening were operated on during the same period. Careful selection of patients who need fine-needle aspiration cytology is demanded of head and neck surgeons, and careful evaluation of indicators is necessary to avoid surgery in the case of benign thyroid disease. PMID- 10695333 TI - [Identification of bacterial strain at each episode of recurrent acute otitis media]. AB - This study was undertaken to investigate whether each episode of recurrent acute otitis media (rAOM) is caused by the same strain of bacteria or different strains at each episode. Seventy infants less than 3-years of age, having experienced rAOM for a period shorter than 8 weeks, were selected and included in the present study. The total number of AOM episodes experienced by this group was 282. At each subsequent episode of AOM, otorrhea and nasopharyngeal swabs were taken for bacterial culture and determination of the MIC for antibiotics. When S. pneumoniae was identified, its serotype, and its pbp, ermAM, and mefE genes were also investigated to determine the bacterial species and strains. S. pneumoniae was the most frequently cultured bacteria with 26 penicillin-sensitive S. pneumoniae (PSSP), 65 penicillin-insensitive S. pneumoniae (PISP), and 50 penicillin-resistant S. pneumoniae (PRSP). H. influenzae was the next most frequently cultured bacteria of which 65 were sensitive to penicillin, 27 were found to be beta-bactamase-negative-ampicillin-resistant (BLNAR) and 17 were found to be beta-bactamase positive. Bacteria cultured from each pair of two successive episodes of AOM were compared as to the identity of the bacteria during the two episodes. In 150 out of 202 pairs (74%), the cultured pathogen was different. In 22 cases in which either PISP or PRSP was the pathogen detected in two consecutive AOM episodes, 15 cases (68%) were found in which the involved strain differed between the two episodes. This study indicates that the pathogen involved in rAOM is likely to differ at each episode of AOM, not only in cases caused by PSSP, but also in those caused by PRSP. PMID- 10695334 TI - [Pharyngeal foreign bodies in infants persisting for two months: two case reports]. AB - In general, patients who suffer from pharyngeal foreign bodies can explain the time and nature of the object which had been swallowed, resulting in easy diagnosis. However, in infants, difficulty in communication makes diagnosis troublesome. Two cases of long-standing pharyngeal foreign bodies were treated at our hospital. Case 1 was a 16-month-old girl complaining of stridor and fever. She had fallen with a toothbrush in her mouth two months before, and was hospitalized in the pediatrics department for one week with upper airway inflammation and dehydration. A toothbrush head was embedded in the back wall of the mesopharynx and hypopharynx. Case 2 was a 10-month-old boy complaining of dysphagia and failure in weight gain. He was hospitalized in the pediatrics department with pneumonia two months before. A PTP (press through pack for medicine) was embedded in the back wall of the hypopharynx. We removed both foreign bodies under general anesthesia. PMID- 10695335 TI - [Snacking behavior among elementary and junior high school students and its relationship to stress-coping]. AB - The aim of the present study was to investigate current problems of snacking behavior and their relationship to stress coping among 1,486 fourth through ninth grade students from 10 elementary schools and six junior high schools. An anonymous self-completed questionnaire was utilized which included items about 1) selection of snack foods, which were classified into healthy, popular, complementary and western-style snacks, 2) problems of snacking behavior, which included external and emotional eating scores, and 3) stress coping scale. The stress coping scale contained two sub-scales; problem-focused and emotion-focused coping. The results were as follows: 1) Students who frequently went without breakfast did not select healthy foods, i.e., fruits and dairy products, but popular snacks, i.e., potato chips, pop corn and sweet beverage. 2) Both external and emotional eating scores increased by age in girls but was not apparent in boys. 3) Students who preferred either western-style or popular snacks showed higher score of external and emotional eating. 4) The score of problem-focused coping was positively correlated with preference for health snacks, but emotion focused coping was positively correlated with external and emotional eating scores. The close relationship between snack food selection and problematic aspects of eating behavior suggests that modification of eating behavior is necessary to develop healthy snack habits in early adolescents. Also, it is interesting that snacking behavior is closely related to stress coping, which suggested the behavioral intervention for healthy eating habit should be included in development of stress-coping skills against various kinds of demands in life. PMID- 10695336 TI - [A field study of health effects of aircraft noise in adults around Komatsu Air Base (1998)]. AB - OBJECTIVE: To ascertain the effects of aircraft noise on complaints about life disturbances and on psychological and physical symptoms in adults around Komatsu air base. METHODS: From April to June 1998, a questionnaire was answered by 203 persons (from 89.8% of families) in 2 areas without aircraft noise, and by 412 persons (from 89.0% of families) in 5 areas with a weighted equivalent continuous perceived noise level (WECPNL) of 75 or more. The questionnaire covered age, gender, work location, noise exposure time, past experience with noisy work and ear disease, complaints about noisiness and life disturbances, and psychological and physical symptoms. Multivariate linear models by JMP (SAS institute Inc.) were applied to analyze the relationships between the above factors including noise level and complaints or symptoms. RESULTS: Subjects in the areas with noise had more complaints about noisiness than 5 years before, although replies were less in areas without noise. Subjects in areas with a WECPNL of 75 or more had more complaints and symptoms than those in areas without noise. Moreover, subjects experiencing higher noise levels gave higher responses for most of complaints and symptoms. The JMP linear models showed that aircraft noise was positively related to complaints and symptoms after taking the effects of age, gender and past experience with noisy work and ear disease into consideration. Higher noise levels and longer exposure times were related more strongly to most complaints and symptoms. CONCLUSION: This study shows that aircraft noise with a WECPNL of 75 or more is related to various complaints and psychological and physical symptoms in adults around Komatsu air base, after considering the effects of age, gender and past experience with noisy work and ear disease. It also shows that higher noise levels and longer exposure times are related more strongly to many complaints and symptoms. The results also suggest that responses to aircraft noise have become more severe than in the past. PMID- 10695337 TI - [Psychiatric distress and related risk factors of family caregivers who care for the demented elderly at home]. AB - The objectives of our study were to assess psychiatric distress of caregivers who had been caring for the demented elderly at home and to examine the association of caregivers' psychiatric distress with putative risk factors. Subjects were 294 caregivers living in Amakusa, Kumamoto Prefecture of Japan, whose spouses, parents or other family members were registered at Amakusa Public Health Center as demented elderly. In 1998, Survey on Caregivers' Mental Health was conducted using the General Health Questionnaire (GHQ12) as a measurement for general psychiatric state of caregivers. Two hundred and eighty-two caregivers responded to interviews with complaints of the following psychological symptoms: feelings of unhappiness (55.7%), of stress (41.8%), insomnia (29.4%) and depressed mood (29.1%). Seventy-six caregivers (27.2%) were identified as being above the cut off point 4 for psychiatric distress caseness. Multivariate logistic regression analysis indicated caregivers' psychiatric distress was statistically associated with caregivers' age, the caregivers' perception of the severity of dementia, the number of years devoted to caregiving at home and perceived financial state. Being 50 to 69 years (OR = 0.37, 95% CI: 0.17-0.81) and being 70 years or older (OR = 0.35, 95% CI: 0.14-0.83) were negatively associated with caseness as compared to being 20 to 49 years. Caseness was positively related to the severity of the elderly's demented state (OR = 6.93, 95% CI: 1.99-24.19), 1 year to 2 years devoted to caregiving at home (OR = 3.26, 95% CI: 1.02-10.38), no family or social support (OR = 2.99, 95% CI: 1.12-7.96) and lower perceived financial state (always OR = 6.99, 95% CI: 2.77-17.64, sometimes OR = 2.41, 95% CI: 1.19-4.85). Reduction of caregivers' psychiatric distress is important for not merely the enhancement of quality of care for demented elders and caregivers' life but for the prevention of elder abuse or neglect. Our study suggests that a comprehensive assessment and intervention program is needed for the demented elderly and their caregivers, in order for caregivers to be able to adapt well to the challenges of caregiving and find personal meaning through caregiving. PMID- 10695338 TI - [Evaluation of the operation of supporting programs for social activity of the elderly by city, town, village governments in Japan]. AB - OBJECTIVES: The aim of this study is to clarify the operative situation of the supporting programs for social activity of the elderly by city, town, village governments in 1997. METHODS: We conducted a study of 3,255 of cities, towns and villages in Japan using a mailed questionnaire, which had been developed to assess the activities of supporting programs for social activity of the elderly. The questionnaire asks government officers whether they had each of the 33 programs in 1997. Each one of these programs belongs to one of the four different aspects of social activities: 1. employment, 2. social participation/volunteering, 3. education/training, 4. individual activities. For each program, the frequency of regions having the program were counted in total, as well as for each of four levels on governments scale: 1. ordinance-designated city, 2. city, 3. town, 4. village. The number of programs operated by the municipality were also evaluated. The main results were as follows One thousand six hundred (49.2%) of cities, towns and villages responded to our questionnaire. 1. Four programs, the promotion of clubs for elderly people, class/lecture meetings for the elderly, sporting events/athletic meetings for the elderly, and a respect-for-age congratulatory gift/money, were put into operation by over 80% of cities, towns and village governments. 2. The number of programs in total or for each of the three aspects of social activities except for social participation/volunteering tended to be high for the large scale governments. There were wide gaps in the number of programs among the regions even of the same level of the government scale. 3. The regions having supporting programs for employment consisted mainly of cities. Seventy-five percent of villages had no supporting program for employment. CONCLUSION: The operational situation of the supporting programs for social activity of the elderly by city, town, village governments was clarified. Furthermore, results obtained by this study can be used for self-assessment of operational situations by municipalities and can contribute to the activation of their supporting programs. PMID- 10695339 TI - [Outbreak-like tuberculosis cases in a general clinic with beds]. PMID- 10695340 TI - [Work load analysis for oral care in special nursing homes]. AB - The work load analysis for oral care in special nursing homes. Dental hygienists undertook oral care of 100 person who were confined to special nursing homes in Aichi prefecture, Japan. The time required, the physical load, the psychological burden were analysed. The results were summarized as follows; 1) Oral care by dental hygienist was incorporated as one of the 59 care tasks given in special nursing. 2) The working time for required oral care was 25.7 minutes per resident on an average. 3) In general, speaking to the elderly, recording on an assessment card, and observing conditions were undertaken most frequently. In oral care, tooth cleaning, gargling, cleaning of dentures, wiping of the mouth and advising those caring for the elderly, was performed most frequently. 4) In general, time taken for advising those caring for the elderly, report to the home dentistry, advising the elderly, recording, and evaluation was most common. In oral care by a dental hygienist, tooth cleaning, cleaning of the dentures, cleaning the mouth, gargling, prevention of dryness of the mouth were performed most frequently. 5) In physical activity, raising the person, moving him from the wheelchair to the bed and helping him to lie down were most frequent. In oral care by a dental hygienist, wiping the mouth, inserting dentures, tooth cleaning, cleaning the mouth and removing dentures were most common. 6) The psychological burden when lifting the person, moving him from the bed to the wheelchair and helping him to lie down was the greatest. In burden involving oral care by a dental hygienist, tooth cleaning, removing dentures, inserting dentures and cleaning the mouth were the greatest. PMID- 10695341 TI - [A survey on drug-related service by homehelper in the Japanese home care system]. AB - The purposes of this study are a) to determine whether homehelpers attend to drug related service of elder care-recipients at home and b) to determine what cooperating human resources were utilized. A structured questionnaire survey was conducted in 1997 with 403 homehelpers who provided in home care to dependent elderly person. Of the total, 19% subjects did deal with care recipients' medication. Regarding the type of service, the highest proportion of subjects had assisted by picking up medicine from pharmacy and talking it to the care recipients' home. Results showed that physicians and home visiting nurses were the most depended upon human resources in the home health care system among health, medical and welfare facilities. Results also suggested that many homehelpers are not aware that pharmacists are readily available resources for providing home health care. Hence we conclude that the respondents tend to rely on physicians or home visiting nurses to respond to care recipients care, and pharmacists should be made aware of the necessity of providing the appropriate drug-related information to physicians or home visiting nurses. PMID- 10695342 TI - [Obesity and life style of Japanese school children with Down syndrome]. AB - A questionnaire-based investigation was performed on 325 Japanese school children with Down syndrome ages 6 to 18. Data on height and body weight, eating habits, physical activity for these children were obtained through their parents. Proportion of obese children was higher among these subjects than the average for Japanese children (34.3% and 7.47% respectively, for the ages from 6 to 14). We examined characteristics of eating habits and physical activities between the obese group (obesity index greater than 20% above the average of Japanese school children) and the non-obese group. Obesity started to increase in the obese group around age 7. The obese group tended to have had a greater intake of sweets, juice and total foods in their preschool days, but unexpectedly had been physically more active in their primary school days. PMID- 10695343 TI - [Current view of macrophage recognition/immune response system to microorganisms]. PMID- 10695344 TI - [Mycoplasmal membrane-bound lipoproteins capable of activating macrophages or fibroblasts to induce cytokine production]. PMID- 10695345 TI - [Molecular epidemiology--molecular typing of pathogenic bacteria using pulsed field gel electrophoresis]. PMID- 10695346 TI - [Molecular epidemiology of enterohemorrhagic Escherichia coli]. PMID- 10695347 TI - [Bacterial intercellular communication and environmental adaptation]. PMID- 10695348 TI - [Ecology of Borrelia and molecular epidemiology of Lyme disease]. PMID- 10695349 TI - [Molecular epidemiology of diphtheria re-emerged in Russia]. PMID- 10695350 TI - [Acute pancreatitis: recent advances in understanding its pathophysiology]. AB - This article reviews the recent changes in the understanding of acute pancreatitis pathophysiology emphasizing results deriving from the more detailed comprehension of the local and systemic aspects of the inflammatory process. The authors briefly discuss those theories that have been influencing the basic philosophies of treatment efforts. The role of premature digestive enzyme activation as the principal determinant of the pathoetiology and mortality of this disease has been questioned lately, and the inflammatory explosion has been placed into the center of attention. Simultaneously with the enzyme activation, the pancreatitogenic noxious event rapidly induces the formation of oxygen derived free radicals, activation of the transcription factor NF kappa-B, with consequent citokine production, cellular adhesion molecule upregulation and leukocyte hyperstimulation. Numerous other mediator cascades are activated in parallel, the uncontrolled surge of proinflammatory stimuli, and activity of the effector cells lead to multiple organ failure in severe cases. A genetically determined catastrophe management program is set forth in the acinar cell with pancreatitis associated protein expression and activation of the apoptosis machinery. Therapeutic approaches based on these recent findings are briefly touched upon. PMID- 10695352 TI - [Acute appendicitis in a patient with increased lead absorption]. AB - The authors describe the case history of an acute appendicitis of a patient suffering from increased lead resorption. The operation resulted in complete healing. They draw the attention to the possibility of an acute abdominal disease also in case of abdominal complaints of patients suffering from increased lead resorption. Careful observation of the patient in collaboration with a surgeon is necessary to avoid a dangerous diagnostic error. PMID- 10695351 TI - [Experience with laparoscopic adrenalectomy in 25 cases]. AB - Between July 1997, and April 1999 the authors performed 25 transperitoneal laparoscopic adrenalectomies. Conversion due to bleeding in 3 cases and due to unsuspected malignancy in 1 case was necessary. Complete resection was carried out in 22 cases. In 3 cases where a well circumscribed adenoma was visualized, only enucleation was performed. A new approach for left sided resection was also introduced. In the authors experience laparoscopy is an absolutely suitable intervention for the benign diseases of the adrenal glands. It assures a short, uncomplicated and painless postoperative period for the patients, with the same effectivity and safety compared to the conventional method. PMID- 10695353 TI - [Two familial cases of hereditary multiple exostoses]. AB - Hereditary multiple exostoses is an autosomal dominant disorder. Three different chromosomal loci have been implicated in this genetically heterogeneous disease. The authors describe a family in which 3 generations were affected, there were data about the disease of an already died grandmother, the father and his daughter were investigated by conventional X-ray and the disease was proved. The disease caused only minor complaints. The exostosis of the father's pelvis showed increased isotope uptake during bone scintigraphic examination, the same region exhibited malignant degeneration on MR examination. Regular check-up of the patients is necessary because of the possibility to a malignant transformation in 1-27% of the cases. PMID- 10695354 TI - [The Cracow University]. PMID- 10695355 TI - [In memoriam Dr. Janos Veres, M.D]. PMID- 10695356 TI - [Intravenous injections of tartaric acid sublimate in streptococcus pyogenes and staphylococcus aureus infections. 1899]. PMID- 10695357 TI - [Contribution to the paper "Depiction of the heart on Hungarian medical coins"]. PMID- 10695358 TI - [The clinical course of end stage heart disease in 152 patients qualified for heart transplantation in a four year observation]. AB - The aim of the study was to analyse the clinical course of pts with end stage disease (ESD) in the period of four years. The study population consisted of 152 pts (132 males, 20 females) at the age of 17-66 years (mean = 48.8 year SD = 9.1) primarily qualified to the heart transplantation (HTX). We analysed the ethiology of cardiac failure, the NYHA class of circulation insufficiency, frequency of occurrence of cardiac arrhythmias and conduction system disturbances in 24-hour ecg monitoring, and the pharmacotherapy efficacy. An ischemic ethiology of cardiac failure we found in 102 pts, cardiomyopathy (idiopathic, hypertrophic or postinfectious) in 46 and unoperable valvular disease--in 4. Ten pts were in II NYHA class, 112 in III, and 30 in IV. Left ventricular ejection fraction (echo assessed) ranged from 11% to 40%(mean = 24.9%), LVEDd = 46-111 mm (mean = 80.9 mm), LVESd = 34-83.5 mm(mean = 63 mm). We found IVa class by Lown ventricular arrhythmias (in Holter monitoring) in 38 pts and IVb in 78. Fifty six pts were treated with amiodarone, 10--with beta-blockers and 11 with sotalol. 19 pts were treated by permanent cardiac pacing during the waiting period, 2 ones--by PTCA, 2 -by CABG, three ones--by dynamic cardiomyoplasty, and one--by partial aneurysmectomy. One pt was treated by CABG and automatic cardioverter defibrilator implantation. In 5 cases HTX was delayed because of the positive effect of pharmacotherapy. In assessed period HTX were performed in 64 cases, 31 pts died and 43 are still waiting for the procedure. CONCLUSIONS: During the 4 year period HTX were performed in 42% of waiting pts. Mortality in this group was 38.2%. In 9 pts (5.9%) the alternative methods of surgical treatment were applicable. In 5 pts (3.9) the decision about HTX was delayed because of the positive change of the clinical status. This fact confirms the necessity of the waiting list verification. PMID- 10695359 TI - [Intracranial hemorrhage during anticoagulant and thrombolytic therapy]. AB - Intracranial hemorrhages are the most severe complication of nowadays widely used anticoagulant and thrombolytic therapy. During 34 months we observed 102 cases of intracranial hemorrhages at the Department of Neurology at our hospital. Among them 8 cases occurred during anticoagulant and/or thrombolytic treatment. The most common risk factors were: arterial hypertension, simultaneous use of other drugs with possible interactions and not following the rules of safe therapy. Because of the dangerous complications during this kind of therapy the risk factors have to be considered. PMID- 10695360 TI - [Modifications in treatment of tetanus and prognosis--observations from the Cracow Department of Infectious Diseases]. AB - The assessment of modifications in the tetanus treatment, which included using metronidazol and midazolam instead penicillin and diazepam, was presented. According to our own observations and previous investigations, mentioned above changes in the tetanus therapy improve survival rate, reduce psychiatric disturbances and shorten hospitalisation time. PMID- 10695361 TI - [The clinical value of B waves in the cerebrospinal fluid pressure curve during the lumbar infusion test of patients with hydrocephalus]. AB - There are still many problems in the selection of hydrocephalic patients for the shunt treatment. Many investigations focus to find out a new and reliable prognostic factors including the parameters of lumbar infusion test (LIT). The aim of this study is to evaluate the B waves observed during the LIT, and to assess the prognostic value of the B waves in hydrocephalic patients. 153 patients with communicating hydrocephalus (ventricular index > or = 0.8) were assessed by means of the LIT. B waves were detected in 70 patients (45.8%). Primary assessment was verified by the Fourier analysis. The relationship was analysed between B waves and the resistance to outflow (R), new steady state pressure (Pns) and the standard deviation of the Pns (SDPns). The results of treatment depending on the presence of B waves were assessed in 95 shunted patients (two categories: improved/not improved). Statistical analysis was performed using Statistically 5.1 for Windows. Chi square test, Mann Whitney U test, and the Student T test were applied (p < 0.05). There was statistically significant relationship between B waves and the increasing values of R (p = 0.0002), Pns (p = 0.0002) and SDPns (p = 0.0000). Positive prognostic value of the B waves was found in the group of 95 shunted patients (p = 0.0152). 1. B waves during LIT appear significantly more often in patients with increasing compensation impairment. 2. B waves detected in LIT have a good prognostic value in hydro-cephalic patients. PMID- 10695362 TI - [[35 Years of effort to improve the diagnosis of porphyria]. AB - World history of porphyria is given in brief. The activities of Porphyria Center during period of 25 years have shown how difficult it was to introduce diagnostic measures for this rare and very little known group of diseases. The obstacles encountered during the popularization of the knowledge of porphyria were: minimal information on porphyria, very seldom seen in Poland, therefore rather negative approach of the doctors to these patients,--almost never porphyria was seen in differential diagnostics of abdominal pains or polyneuropathies, herefore diagnosis, if made, was very late, often just before death. From the other side insufficient support for doctors was given by laboratories because of the lack of simple diagnostic tests. Along the 35 years of the work in the Institute of Haematology and Transfusiology the situation was steadily improving but slowly due to technical and economical difficulties. The Center was organized, now well equipped and able to diagnose and differentiate all types of porphyrias. A computerised basis of the collected material consist of 383 families (6000 persons registered). Among them there are nearly 600 acute (who have passed one or more attacks of porphyria) and about 1000 latent cases of porphyria. The mortality which was 52% in the first collected group of patients (1960-1970) has fallen to less than 10% concerning very severe and late diagnosed attacks. In 1996-1998 there were only two deaths during attacks of porphyria in Poland. PMID- 10695363 TI - [Some aspects of non-pharmacologic treatment of hypertension]. AB - Non-pharmacological methods of intervention are often used as individual or complementary forms of antihypertensive therapy. The aim of the study was to assess the level of compliance and acceptance of some life-style modifications in patients with hypertension. 432 hypertensive patients participated in the study. The study consisted of filling in anonymous questionnaires, taking blood pressure, height and weight measurements. Different approaches of nonpharmacological treatment of hypertension are accepted in differing degree. Life-style modifications and blood pressure self-monitoring should be an integral part of antihypertensive-treatment-programme. PMID- 10695364 TI - [Infections with Chlamydia pneumoniae, Helicobacter pylori or cytomegalovirus and atherosclerosis]. AB - It's known that common risk factors of atherosclerosis can explain only 50% of its etiology. In only 40% patients risk factors modification inhibits progression of atherosclerosis. Therefore looking for new risk factors of atherosclerosis is necessary. In recent years the inflammatory-infectious hypothesis of atherosclerosis has been reevaluated. The discovery of heavy infections load in the serum of patients with acute coronary syndromes might suggest a potential immunological mechanism triggered by bacterial proteins. Chlamydia pneumoniae, Helicobacter pylori and cytomegalovirus infections are very common in human population and therefore they are suspected as the main infectious pathogen in the coronary disease. Clinical studies have demonstrated higher anti-Chlamydia and ani-Helicobacter antibody titres in patients with myocardial infarction (60 70% pts), stable and unstable angina (50-60% pts) than in control groups (12-15% pts). Two studies were performed with antibiotic (azotromycin, roxitromycin) influence on the prevalence of acute coronary syndromes after myocardial infarction and unstable angina. These studies have shown statistically significant reduction of the prevalence of acute coronary episodes in follow-up period. PMID- 10695365 TI - [Endogenous mechanisms for protection in myocardium]. AB - New opinions about myocardial adaptation during ischaemia are described in presented article. Myocardial preservation mechanisms in response to ischaemia are divided into metabolic adaptation like preconditioning and hibernation and anatomic mechanisms of adaptation like development of collateral circulation. Adaptation in clinical conditions can be observed in few situations. 1) During repeated in short period of time exercise tests (warm up). Adaptation may express through increasing tolerated ischemic burden, lenghtening time of exercise, increasing of ischaemic threshold and double product during successive exercise tests (predominately second). This phenomenon called warm up is a clinical counterpart of myocardial preconditioning. 2) In study during coronary angioplasty demonstrated that decrease of ST-segment, intensity of thoracic pain and serum lactacidaemia was lower during the second balloon inflation than the first. 4) The example of myocardial adaptation is repeated atrial stimulation. Ischaemic myocardium should be treated like some kind of mosaic of necrosis, hibernation, stunning and normal viability. PMID- 10695366 TI - [Calcium blockers and their their present application]. AB - Use of calcium channel blockers in arterial hypertension and other cardiovascular diseases has increased substantially during the last decade. The biological role of calcium in contractile tissues mechanisms of action of calcium channel blockers and their present application are discussed. PMID- 10695367 TI - [Dysfunction of platelets in uremia]. AB - Both bleeding and thrombosis may complicate the course of chronic uremia. The main cause of bleeding diathesis is a dysfunction of the primary hemostasis. The widespread improvement of conservative and substitutive therapy has considerably ameliorated the frequency of hemorrhagic complications in uremic patients. The aim of the study review modern views on pathogenesis of bleeding diathesis with particular attention paid to the role of platelets, as well as diagnostic and therapeutic possibilities. PMID- 10695368 TI - [Cytokines in the pathogenesis of liver fibrosis]. AB - Cytokines are synthetized and secreted in the liver mostly by Kupffer cells and play a key role in inflammatory processes and immunological response related to liver diseases, which are initiated by hepatocytes damage. This type of signalling between different types of liver cells, can produce contrary reactions. For instance transforming growth factors beta as well as inetrleukins 1 beta, 4 and 6 induce fibrosis. In contrast, transforming growth factor alpha, tumor necrosis factor alpha, interleukins 1 alpha and interferons are inhibitory to fibrosis. Lost of the balance between these stimuli seems to be responsible for activation of nonparenchymal cells, that result in accumulation of extracellular matrix proteins (including collagens) with liver cirrhosis as a clinical effect. Connections between cytokines, cells located in the liver and diseases with hepatic fibrosis are reviewed in this article. PMID- 10695369 TI - [Depression as a risk factor for cancer--is it still a hypothesis or a proven fact?]. AB - The connection between depression and cancer has been described several times. These observations have been made on major depression and on other kinds of depression as well. It is possible that similar genetic defect may play an important role in the development of cancer and major depression. Disturbances in function of endocrine and immunological systems, which have been observed in depression, can be the risk factor for cancer. Large epidemiological research showed that in persons with depression increased risk of cancer diseases, but not every research confirm it. Large disagreement of research results don't permit of sure answer, but the hypothesis is still possible and require incessant research. Development of research methods, particularly a new domain called molecular epidemiology give hope for more precise future answer. PMID- 10695370 TI - [Pneumothorax in cigarette smokers]. AB - From 1.01.1992 to 31.12.1998 ill prisoners were treated because of so called spontaneous pneumothorax. Their age was between 24 to 57, which was 33 in average. All of them were males and heavy smokers. In one case of the coat pneumothorax the treatment was limited only to the breathing exercises, to ten other patients an under-water-seal drainage was applied, in the next case with pneumothorax and clear symptoms of bronchial fistula, the lung expansion was obtained no sooner than after the usage of a suction drainage. The risk of recurrence of the illness was limited by injection of 40% glucose solution through the drain to the pleural cavity to increase adhesions. PMID- 10695371 TI - [Pulmonary artery sling--diagnostic difficulties]. AB - The pulmonary artery sling is a very rare vascular anomaly which consists in improper branching off and course of the left pulmonary artery. This artery presses on the trachea causing significant symptoms on the part of the respiratory system such as dyspnea, stridor or cyanosis. The authors present the case of a 17-month-old boy who had dyspnea incidents for several times while the reason was unknown. It was only after the X-ray examination of the chest with barium in the esophagus was carried out alongside the bronchoscopy, followed by the echocardigraphic and angiocardiographic examinations that the pulmonary artery sling was found to be reason for those incidences. PMID- 10695372 TI - [Bland-White-Garland syndrome co-existing with ventricular septal defect and patent ductus arteriosus--diagnostic difficulties]. AB - A case of a 14 month-old boy suffering from unrecognized Bland-White-Garland syndrome with no typical clinical and electrocardiological features was reported. Diagnostic difficulties were caused by ventricular septal defect and patent ductus arteriosus co-existing with the anomaly of left coronary artery. Death caused by myocardial ischemia occurred after surgical closure of VSD and PDA. PMID- 10695373 TI - Cybernetics and consciousness. AB - This paper is a review of hypotheses of consciousness which arose from application of the theory of information and regulation and the cybernetic theory of mathematical machines in medicine. The author presents these hypotheses on the examples of his own works. PMID- 10695374 TI - [The understanding of disease in the second half of the XIX century and and the beginning of the XX century and postulates of social medicine. Preliminary study]. AB - Representatives of Polish school of the philosophy looked for the answers to the questions: what the medicine should be and how to use the practice of treatment as a tool of the medical knowledge acquirement? The answers to these questions presented in books and in the journal "Krytyka Lekarska" have contributed to the development of Polish medical thought. The tasks imposed on the social medicine, which in the second half of the 19th century formulated new goals for the medicine, got reflected in this thought too. Health and the endeavor to be free from diseases were of primary importance in the theoretical postulates of the social medicine. Achieving fitness, aiming at it, preserving and enhanding good health were the basic objects of the requests addressed to doctors and medicine. At the same time therapeutic methods offered were often ineffective because the medical activity was focussed on diagnosing and understanding the nature of the disease rather than its treatment. PMID- 10695375 TI - [Scientific medical faculty at the Jagiellonian University from the the Silesian region in 1919-1939]. AB - The article refers to the bonds of the Silesian medicine with the Faculty of Medicine at the Jagiellonian University in interwar period. Fiftieth anniversary of the Silesian Academy of Medicine occurring in 1998 is a chance to recall these relationships. There is a special attention paid to graduates from the Jagiellonian University coming from historical Silesian who graduated from the University, who got doctor's certificates and University degrees. Some of them stayed at the University taking over high ranks, others came back to Silesia, where as high qualified physicians created medical personnel of manufacturing industry of Silesia. PMID- 10695376 TI - [Nonmyeloablative allogeneic hematopoietic stem cell transplantation: minitransplantation]. AB - Allogeneic hematopoietic stem cell transplantation (alloHCT) is considered as a treatment of choice for many malignant hematologic disorders and genetic diseases. Unfortunately toxicities of conventional alloHCT remain a major limitation to successful application of the procedure. A radically new approach for alloHCT has been developed. Nonmyeloablative preparative regimen allows to establish mixed hematopoietic chimerism after alloHCT. A state of stable mixed chimerism may represent a starting point for induction of full donor derived hematopoiesis. A published results of several clinical trials have confirmed potential benefits of this new approach such as less procedure--related toxicity, protection from severe acute GVHD (graft versus host disease), lower TRM (transplant related mortality). Intensive investigations are done to replace in the future pretransplant chemotherapy and/or radiation by nontoxic anti-T-cell agents. These include antibody to the T-cell receptor alpha beta and blockers of T-cell costimulation (e.g. CTLA4lg). PMID- 10695377 TI - [Early results of treating pituitary adenomas by means of transcranial and trans phenoidal approach]. AB - Among the methods of the surgical approach to the pituitary tumours, transsphenoidal (TS) route replaced transcranial (TC) route in the majority of patients. The authors present the development of the surgical treatment of pituitary tumours, and present the early results of treatment by means of both methods of surgery based on the experience of the Department of Neurosurgery in Krakow. Consecutive 45 cases of pituitary adenomas treated by means of TC route (1989-1998)--I group, were compared to 57 cases operated on by means of TS approach (1996-1999)--II group. The other tumours and re-operated patients were excluded from the study. The following factors were compared: the size of treated tumours, the radicality of tumour excision (subtotal, partial), the presence of the massive hypothalamic dysfunction, the presence of diabetes insipidus, changes in visual field, early results of treatment (good, bad, surgical deaths). Using transsphenoidal approach significantly more tumours were removed subtotally (except of large tumours), massive hypothalamic dysfunction was eliminated, surgical deaths were reduced, and the chance for the improvement of the visual field increased. In the last years more patients with small tumours were treated, and less with medium-sized. Percent of patients with large tumours remains unchanged. The prospective study should be performed to define the proper planning of surgical treatment of large and giant tumours, including two-stages procedures. PMID- 10695378 TI - [Intraoperative manometry in laparoscopic antireflux procedures --indications, methods and clinical results]. AB - The aim of the study is evaluating the efficiency of intraoperative manometry during laparoscopic Nissen fundoplication and its ability to prevent postoperative complications. METHOD: Sixteen patients with Gastroesophageal Reflux Disease were included in the study. Clinical examinations, x-ray, endoscopy, pH-metry, and manometric studies were performed before, and 3-6 m.o. after surgery. Fourteen patients were undergoing Nissen fundoplication, and two "floppy Nissen" fundoplications due to the specific preoperative manometric indications. RESULTS: Postoperatively the mean proportion of time at pH < 4.0 on pH-metry decreased from 188 min. (range 96-263) to 8.5 min. (range 2-25). Mean number of reflux episodes significantly lowered after fundoplication from 18.9 (range 2-36) to 0.5 (range 0-3). Gastroesophageal junction mean pressure measured postoperatively reached 24.7 mmHg, and was significantly higher than preoperatively (8.9 mmHg). Mean length of LES increased from 1.2 cm (range 0.8 2.5) to 3.6 cm (range 2.4-4.6) postoperatively. CONCLUSIONS: Laparoscopic Nissen fundoplication assisted by the simultaneous continuous intraoperative manometry is feasible and effective procedure. Continuous LES pressure monitoring during laparoscopic fundoplication with simultaneous computer-video assisted display can be advised as an objective method of intraoperative evaluation of antireflux mechanism. PMID- 10695379 TI - [Determination of blood cell count and spleen size in patients with liver cirrhosis submitted to endoscopic sclerotherapy for esophageal varices]. AB - Each of the 24 patients with bleeding oesophageal varices related to the liver cirrhosis was submitted to 3-12 endoscopic sclerotherapies. During one year of follow-up blood cell counts including erythrocytes, white blood cell, neutorophiles and platelets, the size of spleen on ultrasound were assessed. After a year we noticed marked, statistically significant decline in white blood cells, neutrophiles and platelets counts. The size of the spleen remained unchanged. We may conclude that repeated sclerotherapy of oesophageal varices does not contribute to the rise in blood pressure within the portal system while decrease in blood cells count might be attributed to deterioration of the liver function. PMID- 10695380 TI - [Radionuclide diagnosis of tricuspid regurgitation as a method of estimating the significance of tricuspid insufficiency with concurrent mitral valve disease]. AB - A mitral valve replacement without simultaneous correction of a concomitant tricuspid regurgitation aggravates remote postoperative results. Nowadays diagnostics of a degree of tricuspid regurgitation bases on semi-quantitative methods, which are not unequivocal criteria of a significant tricuspid insufficiency. The aim of the study was to investigate diagnostic usefulness of a radioisotopic method of determination significant tricuspid insufficiency. The study group consisted of 35 patients with rheumatic mitral valve disease and tricuspid regurgitation (30 females, 5 males) at a mean age of 55 years qualified for operative treatment. Physical and noninvasive examinations were performed in all patients: chest X-ray (relative heart volume--RHV) and echocardiographicy (tricuspid regurgitation and right ventricle pressure). Final determination of a significant tricuspid insufficiency based on intraoperative diagnosis. The radioisotopic method relies on first pass technique with a determination of a tricuspid regurgitation index (TRI) and a right ventricular ejection fraction. Intraoperatively the patients were divided into two groups: with significant tricuspid regurgitation--21 patients and without--14 patients. Statistically significant differences, considering clinical and echocardiographic assessment between the two groups were noticed. The TRI index did not differentiate two groups. Noninvasive parameters that could affect diagnosis of significant tricuspid regurgitation were proved by a logistic regression analysis. Among them the TRI Index could have a separate value. CONCLUSIONS: Presented radioisotopic method of determination a degree of tricuspid regurgitation with the new TRI Index is of value in diagnosing significant tricuspid insufficiency when assessed with other noninvasive parameters. Estimation of a clinical usefulness of the method needs further investigation and bigger study group. PMID- 10695381 TI - [Evaluation of noradrenaline and atrial natriuretic hormone concentration in blood serum of pharmacologically untreated patients with chronic heart failure]. AB - Heart failure is a condition of increased neurohormonal stimulation the level of which is affected by both the rate of clinical advancement of the disease and by the manner of its treatment. Hence the work of determining the level of the neuroendocrine activation of the plasma within the range of the adrenergic system and of the atrial natriuretic hormone in not pharmacologically treated patients with mild and moderate heart failure. Consistently investigated were the concentrations of noradrenaline and of the atrial natriuretic hormone in 9 patients (mean age 66.3 years) and the results of the determinations were compared with data of the age approximate group of healthy persons. Then shown, too, could be the concentration increase of noradrenaline and of the atrial natriuretic hormone (both comparisons p < 0.001) over the values that are found in healthy people. PMID- 10695382 TI - [The value of evaluating tumor markers: CA 15-3 and ferritin in blood serum of patients grouped as "high risk" for breast cancer]. AB - We studied 127 patients. 35 patients (group I) with mastopathy disease (average 44.3 years old) included 9 patients with a breast cancer family history (subgroup I A), 26 patients without a breast cancer family history (subgroup I B) and 92 patients (group II) with a breast cancer of I-IV grade (average 49.25 years old). The results of the study revealed statistically significant higher level of CA 15 3 in serum of patients with mastopathy compared to patients with a breast cancer of I grade (p < 0.05). CA 15-3 level in patients with a breast cancer of III, IV grade in serum, was considerably higher and there was significant difference comparing to the patients with a breast cancer of I, II grade (p < 0.001) and patients with mastopathy disease. There were also higher average levels of CA 15 3 in patients with mastopathy disease with a breast cancer family history (subgroup I A) compared to patients without an oncological risk (subgroup I B), however there were not significant differences. There were also higher average levels of ferritin revealed in patients with a breast cancer of III, IV grade and which shows significant difference between patients with breast cancer of I, II grade and with mastopathy patients (p < 0.001). It seems that determination of CA 15-3 and ferritin in patients of a high risk group of a breast cancer could be a useful diagnostic tool for early determination of a breast cancer. PMID- 10695383 TI - [Telomeres and telomerase in leukemias]. AB - There is increasing evidence that telomere shortening both in vitro and in vivo is the clock that counts cell divisions and determines the onset of cellular senescence. Cells overcome the normal senescence mechanism by stabilising telomere length; probably due to the activity of telomerase activity that specifically elongates telomeres. Most human primary tumors contain telomerase, while the cells of most normal tissues lack this activity. Normal haematopoietic cells express telomerase activity. This review is a discussion of utility of telomere length and telomerase activity measurements in the diagnostics and prognosis of leukaemia as well as the potential value of antitelomerase therapy. Results of telomere lengths measurements in young recipients of allogenic transplants are also reported. PMID- 10695384 TI - [Restenosis after percutaneous transluminal coronary angioplasty and its prevention]. AB - The high prevalence of restenosis is considered as a main problem concerning transluminal coronary angioplasty--known and popular invasive method of treatment of coronary heart disease. Mechanisms responsible for vascular wall healing and- also--restenosis as well as the role of cells and its mediators are discussed. The prevention of restenosis is a big challenge for contemporary medicine. The investigations concerning methods which theoretically seem to be justified are being conducted. Unfortunately the results of many trials are unconvincing. At present the most important ways of prevention of restenosis are stent implantation and antiplatelet drugs. PMID- 10695385 TI - [Genetic aspects of primary hyperparathyroidism]. AB - The results of the last decade genetic research in the field of oncogenesis of sporadic and familiar parathyroid tumors are summarised. The review of studied focuses on two main topics: multiple endocrine neoplasis syndromes as well as sporadic parathyroid adenomas and carcinomas. The current knowledge of genetic factors associated with the development of these lesions is presented. PMID- 10695386 TI - [Retrosternal parathyroid gland cystic neoplasm as a cause of primary hyperparathyroidism]. AB - Parathyroid carcinoma is a rare cause of primary hyperparathyroidism (1-2%). In this paper a case of parathyroid cancer of the right inferior parathyroid gland cyst (5 cm in diameter) localised mostly retrosternally is presented in a patient suffering from severe hypercalcaemia (Ca--3.7 mmol/l, Ca(++)--1.8 mmol/l), severe bone pains and weakness, huge osteoporosis with following L5 compression fracture and recurrent nephrolithiasis. PTH blood level before surgical treatment was 1243 pg/ml (Norm: 10-60 pg/ml). In the preoperative ultrasonography of the neck, a lesion was found, but it was considered to be a cyst in the inferior pole of the right thyroid lobe. In a fine-needle biopsy from the lesion-colloid mass without any cells was found. In a 99mTc-MIBI scintigraphy of the neck a suspicion of focus lesion in the right superior parathyroid gland was made, which was not confirmed intra-operatively. Parathyroid carcinoma was diagnosed intraoperatively in histological examination. 'En block' resection of the retrosternal parathyroid tumor and right thyroid lobe was performed from the jugular incision. Any enlarged jugular lymph nodes were not found. In a postoperative period a transient hypocalcaemia was present, which disappeared after pharmacological treatment. During 32 months of follow-up neither no features of local recurrence nor distant metastases were found. Calcium level in blood was normal, PTH 216.0 pg/ml, severity of bone pains and weakness lessened, osteoporosis in a course of pharmacological treatment--with no further progress. PMID- 10695387 TI - [The coexistence of an intracranial tumor and a positive epidemiologic history of Lyme borreliosis as the reason for diagnostic problems--case report]. AB - The authors present the case of a patient treated in the Department of Infectious Diseases at CMUJ in Cracow. The patient's full clinical picture suggested the possibility of the development of neuroborreliosis and disguised the symptoms of a developing intracranial tumor. Neuroborreliosis was suspected due to epidemiologic history (a tick bite, erythema migrans), general symptoms (fatigue, hypersomnia, apathy, dysmnesia, concentration disorders) and neurological symptoms, seropositive tests for Borrelia burgdorferi in serum and cerebrospinal fluid (IgG), increased protein concentration in cerebrospinal fluid. Owing to the fact that the serologic criteria of neuroborreliosis were not fulfilled, and other symptoms (loss of consciousness) appeared, CT was done. The CT showed the presence of a tumor in the longitudinal fissure of the brain, which, after intraoperative and histopathological examination, was defined as meningioma. PMID- 10695388 TI - [Two cases of rare abdominal hernias]. AB - Two cases of rare abdominal hernias were presented: sciatic and obturator. Both presented cases confirm diagnostic difficulties, often leading to delay in therapy. The best way to reduce mortality in these patients seems to be the increase of number of preoperative diagnoses. In obturator hernia's diagnosis the attention is payed to Romberg's symptom, although in sciatic--to symptoms of sciatic nerve irritation or anuria. All diagnostic doubts should be solved with the help of herniography or pelvic CT. PMID- 10695389 TI - [A Polish physician's impressions from the Viennese Clinic of Winternitz]. AB - The achievements of Vincenz Priestnitz, Sebastian Kneipp and Wilhelm Winternitz for the development of hydrotherapy are presented at the beginning. The professional and scientific activities of Winternitz are strongly pointed out. Eugeniusz Piasecki (1872-1947), the Polish physician and theorist of physical education impressions from Vieneese Clinic of Winternitz are depicted widely. The role of Winternitz in formation of modern hydrotherapy in Polish territories is presented finally. PMID- 10695390 TI - [How a Warsaw physician helped Sienkiewicz in his literary debut]. AB - This article is on the borderline between history of medicine and history of literature. The life and works are presented of Konrad Dobrski (1849-1915), with description of his works in Warsaw and scientific visits to Vienna. He was a well known Warsaw's physician, especially in laryngology and pulmonology. His friendly relations with Henryk Sienkiewicz, a famous Polish writer, in adolescence are described in more details. The Dobrski's participation in origin of first Sienkiewicz's novel "Na marne" is presented with full particulars. The Jozef Ignacy Kraszewski's assistance in this matter is also depicted. PMID- 10695391 TI - [Analysis of pulmonary function in leukemia patients after bone marrow transplantation: effects of prior chemotherapy]. AB - To evaluate the effects of prior chemotherapy on pulmonary function after bone marrow transplantation(BMT), pulmonary function tests were performed prior to and after BMT on 7 acute leukemia (AL) and 13 chronic myelogenous leukemia (CML) patients given with CY (60 mg/kg x 2 days), total body irradiation (3 Gy x 4 days, 10 cGy/min), and CyA plus short-term MTX. No patient had graft-versus-host disease or lung complications. Pulmonary function after BMT did not deteriorate in the AL patients; however, both %Vital Capacity(%VC) and DL/VA decreased significantly in the CML patients (%VC before BMT: 112.1 +/- 11.5%, after BMT: 93.7 +/- 9.4%; DL/VA before BMT: 79.2 +/- 14.6%, after BMT: 54.1 +/- 10.6%). Although prior regimens of busulfan (BU) or interferon (IFN) were equal risk factors for decreased %VC after BMT, decreases in DV/VA were more significant in CML patients who received IFN. CML patients, especially those who have received BU or IFN, should be carefully monitored for pulmonary function to prevent respiratory failure after BMT. PMID- 10695392 TI - [Low-dose prednisolone therapy for idiopathic thrombocytopenic purpura]. AB - Prednisolone (PSL) is widely used for the treatment of idiopathic thrombocytopenic purpura (ITP). We compared the effects of a relatively low dose (0.5 mg/kg/day, LD group) of PSL and the conventional dose (1.0 mg/kg/day, CD group) on 59 ITP patients. Twenty-six patients were treated with low-dose PSL, and 23 patients with the conventional dose. No statistically significant difference was observed in the complete remission rates for the LD group (35%) and the CD group (39%). However, the mean duration of hospitalization was significantly (p < 0.001) shorter for LD group patients than for patients in the CD group (20 days versus 50 days, respectively). In conclusion, low-dose PSL may be as effective as the conventional dose and capable of reducing the cost of hospitalization, thus, improving the quality of life for patients with ITP. PMID- 10695393 TI - [A retrospective study of the length of hospitalization following allogeneic bone marrow transplantation]. AB - To estimate the length of hospitalization following bone marrow transplantation(BMT), we conducted a retrospective study of 190 patients who had received allogeneic BMTs at our institution. By our criteria, patients were considered ready for discharge if they were afebrile, did not need intravenous chemotherapy or blood transfusions more than 2 times per week, had maintained these conditions for 1 week or more, and also had no medical history of hepatic veno-occlusive disease, grade-II-or-higher graft-versus-host disease, interstitial pneumonitis, or severe hepato-renal dysfunction. The median length of hospitalization was 108.5 days. Of 82 patients who satisfied our discharge criteria by their 70th hospital day, 10 experienced mild complications during the next 30 hospital days. Of 89 patients who were considered ready for discharge by the 40th hospital day, 30 and 38 experienced complications during the next 30 and 60 hospital days, respectively, and 16 required emergency treatment. No significant baseline characteristics distinguished the patients who experienced complications from those who did not, either after 40 or 70 hospital days. This compounded the difficulty of predicting the development of complications in patients who satisfied our discharge criteria. Although management on an outpatient basis should be safe and feasible for BMT patients who meet our discharge criteria by the 70th day of hospitalization, caution is advised for early discharges after only 40 hospital days. PMID- 10695394 TI - [Varicella-zoster virus infection after hematopoietic stem cell transplantation]. AB - Of 264 patients aged 15 years or more who underwent hematopoietic stem cell transplantation between 1989 and September 1998 at the Tokyo Metropolitan Komagome Hospital, 47 were infected by the varicella-zoster virus (VZV). In 2 patients, visceral disease preceded cutaneous dissemination. One of these patients exhibited gastrointestinal symptoms followed by disseminated skin rash 6 days later. In the other patient, epigastralgia developed and was followed by seizures secondary to meningitis; the appearance of a skin rash 5 days after these initial symptoms yielded the diagnosis. Early diagnosis and treatment of VZV infection are important, especially for patients who present with visceral symptoms suspected to be due to VZV. PMID- 10695395 TI - [Bone marrow transplantation-associated thrombotic microangiopathy manifested by visual disturbance]. AB - In October 1996, a 26-year-old woman was given a diagnosis of acute myeloblastic leukemia, FAB subtype M1. Treatment with combined chemotherapy achieved a complete remission (CR). In May 1997, the patient received an allogenic bone marrow transplant (BMT) from an HLA-identical sibling donor. Cyclosporine (CsA) and short-term methotrexate were given for graft-versus-host disease (GVHD) prophylaxis. Successful engraftment was obtained and signs of acute or chronic GVHD never developed. Five months after BMT, the patient experienced low-grade fever and blurred vision. Retinal examination demonstrated intraretinal hemorrhages, cotton-wool spots, and retinal detachments, which were presumably attributable to multiple thrombosis of retinal microvessels. The patient also exhibited hemolytic anemia with red cell fragmentation, thrombocytopenia, elevated lactate dehydrogenase, and renal impairment, and was thus given a diagnosis of BMT-associated thrombotic microangiopathy (BMT-TM). Discontinuation of CsA and administration of ticlopidine and prednisolone induced successful recovery from BMT-TM. Three months after the onset of BMT-TM, however, the patient experienced generalized clonic-tonic seizures with consciousness loss. Single-photon-emission computed tomography revealed blood-flow disturbances in the brain, suggesting the recurrence of microthrombosis. Accordingly, multiple transfusions of fresh frozen plasma were administered together with dipyridamole and aspirin. The patient gradually recovered and remained asymptomatic through the following 13 months. Currently, early diagnosis of BMT-TM is considered to be difficult. We suggest that careful examination of the ocular base may be useful for the early detection of BMT-TM. PMID- 10695396 TI - [CD56-positive adult T-cell leukemia manifested by abnormal lung shadows]. AB - A 54-year-old woman was admitted to Juntendo Izunagaoka Hospital on Aug. 29, 1998, after experiencing cough and fever for 19 days. Chest X-ray films disclosed infiltrates in the left lung field. The abnormal lung shadows progressed despite antibiotic therapy, and enlargement of superficial lymph nodes and hepatosplenomegaly developed. Peripheral blood examination disclosed cleaved lymphoid cells without granular cytoplasm. Anti-HTLV-I antibody titer was x320, and the monoclonal integration of HTLV-I provirus was confirmed by Southern blot analysis. Surface marker analysis of lymph node cells was positive for CD2, CD3, CD4, CD5, CD56, and HLA-DR. The above results yielded a diagnosis of adult T-cell leukemia. LSG-4 therapy alleviated the lung infiltrations and dyspnea. This case was considered unusual because of the expression of the natural killer cell marker CD56 on leukemic cells and the presentation of abnormal lung shadows possibly due to leukemic cell infiltration. PMID- 10695397 TI - [A boy with summer onset paroxysmal cold hemoglobinuria induced by Donath Landsteiner antibody with anti-I specificity]. AB - We encountered 4-year-old boy who developed paroxysmal cold hemoglobinuria (PCH) in the middle of August. He was admitted due to progressive jaundice and pallor following fever and nausea. Laboratory data revealed severe anemia, increased serum indirect bilirubin and LDH, and decreased serum haptoglobin. Direct/indirect Coombs tests were positive. These findings yielded a diagnosis of autoimmune hemolytic anemia. Serological test for syphilis was negative. The patient's symptoms and signs promptly subsided after treatment with prednisolone, which was started on the 2nd hospital day. The patient has been in a disease-free state for 16 months without any medication. After discharge, he was finally given a diagnosis of PCH because a Donath-Landsteiner test was positive. The antibody belonged to an IgM subclass and showed anti-I specificity. Considering that development of PCH is common in winter, this case was unique because it developed in August. We speculated that exposure to a cool air-conditioned atmosphere prior to hospitalization and the cooling mechanism of the body after admission were involved in the summer onset of PCH and its prolonged clinical course. PMID- 10695398 TI - [Sustained long-term complete remission in an elderly aplastic anemia patient after cessation of combined therapy consisting of granulocyte-colony stimulating factor and erythropoietin]. AB - An 83-year-old woman received a diagnosis of moderate aplastic anemia in November 1990. Immunosuppressive therapy consisting of anti-lymphocyte globulin combined with high-dose corticosteroids was effective until pancytopenia developed in August 1993. The patient was hospitalized again and recurrence of aplastic anemia was diagnosed on the basis of hematologic findings, including RBC 129 x 10(4)/microliter, Hb 5.5 g/dl, Ret 23,200/microliter, WBC 2,200/microliter with 27% neutrophils, platelets 2.2 x 10(4)/microliter, and hypoplastic bone marrow. Recombinant human granulocyte-colony stimulating factor (G-CSF) of 125 micrograms/day combined with recombinant human erythropoietin (EPO) of 6,000 U/day were started in November 1993. The doses of G-CSF and EPO were increased to 250 micrograms/day and 12,000 U/day, respectively. We stopped combination therapy in March 1995, after trilineage hematopoietic cell recovery had been achieved. Complete recovery in peripheral blood was sustained for more than 2 years despite the termination of G-CSF and EPO therapy. Combination therapy with G-CSF and EPO may be safe and effective for elderly patients with aplastic anemia when the choice of therapy is limited. PMID- 10695400 TI - [Histiocytic necrotizing lymphadenitis (Kikuchi-Fujimoto's disease) accompanied by hemophagocytosis and salivary gland swelling in a patient with systemic lupus erythematosus]. AB - After 2 years of steroid therapy that had effectively controlled her systemic lupus erythematosus, a 37-year-old woman presented with fever, erythema (face, upper chest), and low CH50. Increased oral steroid (prednisolone from 15 mg to 40 mg) and intravenous methylprednisolone (mPSL) (80 mg for 3 days) alleviated these symptoms except for the fever. Subsequently, the patient's fever worsened and leukocytopenia, abnormal liver function, lymphadenopathy (neck, axilla), and salivary gland swelling developed. Lymph node histology revealed features characteristic of Kikuchi-Fujimoto's disease (KFD). Laboratory examinations showed WBC 600/microliter, Hb 9.5 g/dl, platelets 90,000/microliter, GOT 766 IU/l, GPT 646 IU/l, LDH 4,228 IU/l, TG 1,622 mg/dl, and ferritin 6,330 ng/ml. Serum interferon gamma was also elevated (673 U/ml). Because a bone marrow smear revealed hemophagocytosis, mPSL pulse therapy (1 g for 3 days) was started for treatment of hemophagocytic syndrome. The fever promptly disappeared, and the patient's clinical symptoms resolved within 2 weeks. The abnormal laboratory data related to KFD and hemophagocytosis returned to normal within 4 weeks after the initiation of mPSL pulse therapy. We speculated that the hemophagocytosis and salivary gland involvement in this patient were also symptoms of KFD. This case indicated that corticosteroid pulse therapy is effective for KFD with serious clinical symptoms. PMID- 10695399 TI - [Two cases of orbital non-Hodgkin's lymphoma presenting with hemophagocytic syndrome]. AB - We report 2 cases of orbital non-Hodgkin's lymphoma (NHL) with hemophagocytic syndrome (HPS). Patient 1 was a 64-year-old man with a diagnosis of peripheral T cell lymphoma originating in the right orbita (clinical stage: IV B). Epstein Barr virus DNA was demonstrated in tissue specimens by polymerase chain reaction. Laboratory findings on admission were WBC: 4,700/microliter, Hb: 12.1 g/dl, Plt: 14.6 x 10(4)/microliter, LDH: 951 IU/l, sIL-2R: 2,553 IU/ml, and ferritin: 5998.1 ng/ml. Patient 2 was a 73-year-old man with a diagnosis of diffuse large B-cell lymphoma originating in the right orbita (Clinical stage: IV B). Laboratory findings on admission were WBC: 9,100/microliter, Hb: 7.7 g/dl, Plt: 15.4 x 10(4)/microliter, LDH: 1,043 IU/l, sIL-2R: 10,090 IU/ml, and ferritin: 2079.3 ng/ml. Both patients had high-grade fever and extremely high serum cytokine levels. Bone marrow aspiration disclosed many histiocytes with hemophagocytosis. In both cases, combined chemotherapy was transiently effective, but patient 1 died of relapse of HPS and patient 2 of cerebral bleeding. Orbital non-Hodgkin's lymphoma with HPS is rare. These cases were interesting in terms of the relationship between HPS and the primary site of lymphoma. PMID- 10695401 TI - [beta-thalassemia minor diagnosed in a patient with chronic myelogenous leukemia during hydroxyurea therapy]. AB - A 55-year-old man was admitted to our hospital because of leukocytosis and microcytic anemia with hypochromia, target cells, and increased levels of hemoglobin A2 and hemoglobin F. The results of a gene analysis yielded a diagnosis of chronic myelogenous leukemia and beta-thalassemia minor. A gradual increase in hemoglobin was observed during hydroxyurea therapy, which was performed over a 12-week period. This increment appeared to be due to suppressed production of myeloid cells. It was been reported that hydroxyurea increases total hemoglobin due to increased hemoglobin F synthesis in patients with beta thalassemia. However, hydroxyurea had no clear influence on hemoglobin concentration in this case. PMID- 10695402 TI - [An aplastic anemia patient died of severe anemia who refused transfusion]. AB - A 41-year-old woman who had been given a diagnosis of aplastic anemia 14 years before was admitted because of recurrence of the disease. Despite therapy, the anemia progressed gradually. The patient refused blood product transfusions for religious reasons. Angina pectoris-like chest pain without ischemic changes on electrocardiograms appeared at a hemoglobin concentration (Hb) of 1.6 g/dl. The patient died of heart failure at Hb 1.5 g/dl. Autopsy showed enlargement of the heart, fatty changes in the myocardium and liver due to chronic hypoxia, and no changes in coronary arteries. PMID- 10695403 TI - [Essential thrombocythemia in transformation from myelodysplastic syndrome to acute myeloid leukemia with inv(3) after treatment for gastric cancer]. AB - In March 1990, a 61-year-old man was given a diagnosis of essential thrombocythemia with a normal karyotype and subsequently treated with hydroxyurea. In November 1995, he underwent surgery for gastric cancer with thereafter received tegafur/uracil for 2 years. Refractory anemia with excess of blasts in transformation and chromosomal abnormalities including -5, -7, 20q developed in August 1998. Combined chemotherapy with daunorubicin, cytarabine, mercaptopurine, and prednisolone, had only limited effectiveness. Acute myeloid leukemia was finally diagnosed in October 1998, and chromosomal analysis disclosed inv(3) in addition to -5 and -7. The appearance of inv(3) might be related to leukemic transformation of hematopoietic stem cell disease with an increase in the number of megakaryocytes and platelets. PMID- 10695404 TI - [My recent daily life especially on hemo dialysis]. PMID- 10695405 TI - [Nation-wide survey for the treatment with cyclosporin A of interstitial pneumonia associated with collagen diseases]. AB - OBJECTIVES: This study was performed to investigate the efficacy and safety of cyclosporin A (CsA) for the treatment of interstitial pneumonia (IP) associated with collagen diseases in Japan. METHODS: Questionnaires were sent to 36 hospitals specializing in collagen diseases. RESULTS: Fifty-eight patients (7 polymyositis (PM), 19 dermatomyositis (DM), 7 systemic sclerosis (SSc), 7 rheumatoid arthritis (RA), 2 mixed connective tissue disease (MCTD), 1 systemic lupus erythematosus (SLE) and 1 Sjogren's syndrome (SS), 1 RA + SSc, 2 PM + SSc, 1 DM + SLE, and 10 idiopathic interstitial pneumonia (IIP) with IP were treated with CsA at 14 hospitals. IP was classified into the acute or chronic type. In the PM/DM group (7 PM, 19 DM, 2 PM + SSC, 1 DM + SLE), 65.5% were the acute type. In the other collagen disease group (7 SSc, 7 RA, 2 MCTD, 1 SLE, 1 SS, and 1 RA + SSc) and IIP group, 36.8% and 50% were the acute type, respectively. Mean dosages of CsA were 3.7 +/- 1.3 mg/kg/day for the PM/DM group, 3.0 +/- 1.0 for the other collagen disease group, and 3.8 +/- 4.8 for the IIP group. Oral corticosteroids were administered in combination with CsA in 100, 73.7, and 70% of the patients with PM/DM, other collagen disease, and IIP groups, respectively. CsA was effective for 72.2, 33.3, and 25% of the acute IP cases in the PM/DM, other collagen disease, and IIP groups, respectively. CsA was effective for 50.0, 50.0, and 60.0% of chronic IP cases in the PM/DM, other collagen disease, and IIP groups, respectively. Twenty-three adverse effects were observed, but most of them ameliorated upon withdrawal or reduction of the CsA dose. CONCLUSION: CsA is effective for the treatment of acute type IP associated with collagen diseases, especially PM/DM. To perform a prospective multi-center trial, standards for the recruitment of patients, efficacy assessments, and trial course and treatment should be determined carefully. PMID- 10695406 TI - [Improvement of the maintenance therapy after methylprednisolone pulse therapy- effect of prednisolone combined with immunosuppressants]. AB - OBJECTIVES: We investigated the effect of the combination therapy of prednisolone (PSL) and immunosuppressants after methylprednisolone pulse therapy. METHODS: A protocol of PSL (15-20 mg/day) and mizoribine (150-200 mg/day) after methylprednisolone (mPSL) pulses was used for 2 years to treat 7 patients (PSL + MZB group). Cyclophosphamide (CYC) pulse therapy was added to the combined therapy in 4 patients with severe lupus nephritis. The total dose of predinisolone, and side effects were compared with those in 6 patients who were treated with PSL (30 mg/kg) alone after mPSL pulse therapy (PSL group). RESULTS: No relapses occurred in the PSL + MZB group, although all of 6 patients relapsed in the PSL Group. The total doses of PSL in the PSL + MZB group was about 70% of the PSL Group. There were two patients with Herpes-Zoster infection and one patient with liver dysfunction as side effects, with no differences in the frequency of side effects between the was groups. CONCLUSIONS: Combination maintenance therapy with prednisolone and immunosuppressants after methylprednisolone pulse therapy was effective in preventing relapse. PMID- 10695407 TI - [A case of amyopathic dermatomyositis presenting blister and oral ulcer]. AB - An 84-year old man was admitted to Mitoyo General Hospital because of progressive malaise and edematous erythema on both eyelids (Heliotrope erythema). He also noted blister on his neck as well as erythema on the extensor surface of finger joints (Gottron's sign), elbows and knees. Neither weakness nor pain of his proximal muscle was elicited on physical examination on admission. His blood test disclosed positive inflammatory signs (i.e., mild elevation of ESR and positive CRP) without elevated value of muscle enzymes. Electromyogram showed normal pattern. Infiltration of inflammatory cells was not revealed by histological examination of biopsied muscle. A diagnosis of 'amyopathic dermatomyositis' was made based on these observations. Computed tomography of his chest disclosed interstitial pneumonia spreading over both lower lung fields. Colon fiberscopy revealed a polyp in his descending colon, which was classified into group I on cytological examination. He was treated with two sets of methylprednisolone (mPSL) pulse therapy (500 mg/day, 3 consecutive days, intravenously) followed by 30 mg/day of oral prednisolone (PSL). His skin lesions responded well to the above treatment and the dose of oral PSL was tapered. One month after the initiation of treatment, severe stomatitis as well as a large ulcer beneath his tongue developed accompanying an intractable pain. Mucosal biopsy revealed necrotizing vasculitis in medium-sizedartery at the bottom of ulcer. Another set of mPSL pulse therapy brought a prompt relief of his symptom and prohibited the recurrence of oral lesion. It should be noted that our patient did not fulfill the diagnostic criteria for DM because of the lack of muscular symptoms whereas he had characteristic skin lesions. Regarding the frustration possibly encountered at the time of diagnosing amyopathic DM, both sensitivity and specificity of the skin lesion for the diagnosis of DM were investigated. Moreover, the rarity of blister as a skin manifestation of DM was discussed as well. PMID- 10695408 TI - [Two cases of silicosis exhibiting MPO-ANCA associated disorder]. AB - We reported two cases of silicosis exhibiting MPO-ANCA associated disorder. Case 1 was a 69 year-old man with silicosis and chronic interstitial pneumonia. He was admitted because of fever, dry cough, left chest pain, dyspnea and body weight loss. He was diagnosed as acute exacerbation of interstitial pneumonia, pericarditis and gastrointestinal bleeding. Case 2 was a 67 year-old man with silicosis. He repeated attack of fever, hoarseness, dysphagia and headache. The cell counts of cerebrospinal fluid increased and the thickness of cerebellar tent and left dura mater was observed in the brain MRI. Therefore, he was diagnosed as pachymeningitis and neuropathy of cranial nerves. Both cases were complicated by silicosis and the laboratory findings showed high serum levels of P-ANCA, ANA and rheumatoid factor and inflammatory responses, indicating they were suspected vasculitis. The two cases were treated by steroid and immunosuppressive therapy and had good clinical response. Silicosis may affect multiple organ involvement associated with P-ANCA. PMID- 10695409 TI - [A case of Sjogren's syndrome with retrobulbar optic neuritis and cutaneous vasculitis]. AB - A 52-year-old woman, diagnosed as having Sjogren's syndrome by parotid sialography and lip biopsy after a two years history of recurrent purpuric rashes on her lower extremities, was admitted to our hospital because of visual disturbance in March 1998. On presentation at the department of ophthalmology, her right visual acuity was light perception, and laboratory findings showed elevated levels of antinuclear antibody and anti-Ro/SS-A and anti-La/SS-B antibodies. Cerebrospinal fluid analysis showed mild pleocytosis and elevated levels of total protein and Q-albumin. The IgG-index was within normal level and no oligoclonal band was found. Magnetic resonance imaging showed increased signal intensity at the right optic nerve. After treatment with m-PSL pulse therapy, her visual acuity recovered to 0.08. When prednisolone was gradually tapered to the dose of 30 mg per day, she was transferred to our department because of high grade fever and pancytopenia. She also suffered from palpable purpura in her extremities extending the trunk, whose pathological diagnosis was leukocytoclastic vasculitis. The immunohistochemical examination showed depositions of IgG and C1q. After two additional cycles of mPSL pulse therapies, clinical improvement was achieved. The titers of von Willebrand factor and thrombomodulin correlated with her clinical improvement. Patients with Sjogren's syndrome can develop extra-grandular complications, including neurologic and cutaneus diseases, it is important to understand the role of SS-A-B antibodies in the immunopathogenesis of Sjogren's syndrome. PMID- 10695410 TI - [An autopsy case of aortitis resulting in a tear of the aortic valve due to a rupture of the aneurysm of Valsalva's sinus]. AB - A 43-year-old man was admitted to a hospital because of acute dyspnea and nocturnal orthopnea. Echocardiogram and chest CT showed the dilation of thoracic aorta from the root to ascending portion. On the third hospital day, he died suddenly. At autopsy, the cause of death was indicated to be a tear of an aortic valve due to a rupture of the aneurysm of Valsalva's sinus, followed by acute aortic regurgitation and acute cardiac insufficiency. Histopathological findings of thoracic aorta revealed mesoaortitis, characterized by patchy destruction of the media with a moth-eaten appearance of the medial elastic laminae and a microgranuloma formation, a perivascular mononuclear cell infiltration of the vasa vasorum, and a fibrous thickening of the intima and adventitia. However, there were no abnormalities in main branches of aorta and abdominal aorta, and no systemic vasculitis. This case is a rare one in the clinical course, and may be important to be differentiated from other cases with aortitis, especially Takayasu arteritis and syphilitic aortitis. PMID- 10695411 TI - [The recent progress of the treatment for chronic idiopathic arthritides of childhood]. PMID- 10695412 TI - [Rheumatoid arthritis and human parvovirus B 19]. PMID- 10695413 TI - Nitric oxide--the magic molecule. PMID- 10695414 TI - Cytokines and inflammatory bowel disease. AB - Cytokines are the key mediators of inflammation in the IBD and are focus of renewed interest to plan therapeutic strategies against this disease. However, there are gaps in our knowledge at present and a lot of questions need clear answers. Even with a therapy as specific as anti-TNF antibody, it is not clear if the benefit is attributable to simple binding and clearance of TNF-alpha or to binding on the cell surface and subsequent deletion of the activated macrophage. When a drug appears to be less effective than pre-clinical models suggest, can failures in effectiveness from delivery or dosing the differentiated? The disappointing results of clinical trials with IL-10 is at odds with the prediction of benefit from animal models. It even brings into question the validity of those models as well as the soundness of design of the clinical trials on which efficacy of IL-10 is assessed. Other exciting new methods to treat IBD could be use of monoclonal antibodies to effector T cell molecules (such as CD4 or CD44v7) removal of such cytokine secreting cells (apheresis), antibodies to proinflammatory cytokines (such as TNF-alpha, IFN-alpha, IFN-gamma, and IL-12) or administration of anti-inflammatory cytokines (such as IL-10, IL 11). Challenges in developing new therapeutic strategies include not only identifying novel agents, but also improving the definitions of clinical endpoints and defining efficacy at the biologic level. There is also need to further refine our knowledge about genetic elements and environment initiators to comprehensively manage IBD. PMID- 10695415 TI - Clinical significance of carcinoembryonic antigen in colorectal malignancy. AB - Carcinoembryonic antigen (CEA) assay was performed in 40 patients of histologically proven colorectal carcinoma. The overall incidence of positivity was 72.5%. The incidence increased from 40% in Duke's A stage to 84.6% in Duke's C stage. Similarly the mean CEa levels also increased as the disease advanced i.e. 4.96 ng/ml, 8.07 ng/ml and 12.7 ng/ml in Duke's A, B and C respectively. Cancer with poor prognosis i.e. poorly differentiated and colloid carcinoma, had significantly less rise in CEA values (P < 0.05) as compared to well differentiated carcinoma. There was no relation of CEA values with the gross appearance of the tumour and lymph node involvement. CEA level came down in all the patients after surgery. Based on the postoperative CEA estimation, complete tumour clearance had been achieved in 86.2% of patients. PMID- 10695416 TI - Barrett's oesophagus and oesophageal cancer in Saudi Arabia. AB - BACKGROUND/AIMS: Gastrooesophageal reflux disease (GERD) and Barrett's oesophagus (BE) are presumed to be rare among non-western populations. This retrospective survery determined the prevalence of BE and its associated complications of dysplastic lesions (DL) and oesophageal adenocarcinoma (AD) among an Arabian population. METHODOLOGY: From 2572 patients who had endoscopy in King Fahd Central Hospital (KFCH) Gizan, Saudi Arabia, patients (n = 776) were selected for analysis if they had biopsies of the upper gastrointestinal (UGI) tract. The patients (159 of 776) with biopsy-proven oesophageal lesions were categorized and compared. RESULTS: The relative frequencies of BE, DL and AD in 159 patients were 8(5%), 5(3%) and 16 (10%) respectively. These interpreted to prevalence rates of 0.31%, 1.9%, and 0.62% for the respective lesions. The comparison of the mean age +/- SD (in years) of the patients with BE (59.6 +/- 19.8), DL (66 +/- 16.7) and AD (70.6 +/- 12.2) showed to statistically significant difference. Major symptoms in the patients with BE were dyspepsia (4 cases), hematemesis (2 cases) and dysphagia (2 cases). This profile was not different from that observed in 79 patients with GERD. CONCLUSION: The prevalence of 0.31% in our endoscopy population is at the lower range of the 0.3% to 10% reported in the western world. It is likely that the rate was underestimated by this retrospective survey, in which patients were selectively biopsied. Also, it is probable that the majority of individuals in our population with no or minimal symptoms of GERD do not present themselves or are not referred for evaluation. Despite this limitation, our study confirms the occurrence of BE and its complications among a Saudi population. The incidence of BE may increase with the current changes in the life-style and increase in the life-span of the Saudi Arabian population. PMID- 10695417 TI - Double blind controlled trial of effect of cisapride on gastric emptying in diabetics. AB - AIMS: Diabetic gastroparesis is a common complication seen in 20-50% of patients due to autonomic neuropathy involving vagal supply. Cisapride, a specific gastrointestinal cholinomimetic agent may thus be effective. METHODS: Fifty-one diabetic patients (age 12-65 years) of disease duration > 5 years were assessed for symptomatic gastroparesis, other diabetic complications and glycemic control. Gastric emptying time (GET) was estimated using a solid meal method (99mTc labeled rice based idli) and patients randomized to receive either cisapride or placebo for a period of 2 weeks. Cisapride was administered in a dose of 10 mg TID. GET and symptom scores were reassessed on the therapy after 2 weeks. RESULTS: Twenty nine of 51 (56.8%) patients had gastroparesis. Mean GET in the gastroparesis group was 141 +/- 66 minutes compared to 24.53 +/- 10 minutes in the non gastroparesis group (p < 0.01). GET decreased by 72% amongst the patients who received cisapride compared to 23% in the placebo group (p < 0.001). Symptom scores also improved in the cisapride group; no adverse effects were noted. CONCLUSIONS: Cisapride improves the symptom score and the solid gastric emptying time in patients suffering from diabetic gastroparesis. PMID- 10695418 TI - Assessment of malnutrition in alcoholic and non alcoholic cirrhotics. AB - OBJECTIVE: To study the nutritional status in patients with chronic liver disease using anthropometric techniques. METHODS: A total of 60 cirrhotic patients (30 Alcoholic (AC), 30 Non-alcoholic (NAC) and 30 control (CO) subjects were studied. Nutritional status was assessed using anthropometric measurements such as stature, body weight, body mass index, (BMI), skinfold thickness measurements and mid upper arm muscle circumference. Serum protein, serum albumin and globulin were measured. RESULTS: The skinfold thicknesses were significantly lower in NAC group of patients. In contrast the AC group of patients showed significantly lower mid upper arm muscle circumference values. Both groups of cirrhotic patients showed significantly lower total serum protein and serum albumin levels. CONCLUSION: Body fat is relatively more affected in NAC group of patients and muscle mass is more affected in AC group of patients. PMID- 10695419 TI - Gastric protection against cold restraint stress-induced lesions by amoxycillin in rats. AB - AIMS: Recent investigations have shown that amoxycillin possesses gastric protection properties in addition to its known antimicrobial effects. Therefore, this study was undertaken to investigate the potential gastric protection effects of amoxycillin and to determine its possible mechanism(s) of action in rats. METHODS: The cold restraint stress model was used to produce gastric mucosal lesions. The gastric secretion studies were undertaken by using Shay's pylorus ligation technique. The antioxidant effect was studied by luminol dependent chemiluminescence technique in vitro. RESULTS: Amoxycillin dose-dependently prevented cold restraint stress-induced mucus depletion and afforded protection. It inhibited indomethacin-stimulated gastric acid secretion with a high dose without affecting basal secretion. Furthermore, amoxycillin dose-dependently inhibited the phorbol myristate acetate-stimulated luminol-dependent chemiluminescence responses of isolated human poylmorphonuclear leukocytes in vitro. CONCLUSIONS: These results suggest that mechanisms of gastric protection effects of amoxycillin may include inhibition of stimulated acid secretion, prevention of depletion of mucus and antioxidant properties. PMID- 10695420 TI - Effect of hepatitis C virus coinfection on liver function of patients infected with HIV. AB - AIMS: Both the human immunodeficiency virus (HIV) and Hepatitis C virus (HCV) share similar routes for their transmission, encouraging the possibility of a coinfection with the two viruses in some individuals. We wanted to find out the prevalence of HCV in patients infected with the HIV virus. METHODS: We tested 57 HIV positive patients for anti-HCV antibodies. Liver functions tests, ultrasound abdomen and upper gastrointestinal endoscopy were carried out in the anti-HCV positive patients. RESULTS: Coinfection with HCV was seen in 5.3% of patients. Of these 2 had acquired the infections through blood transfusions and one through heterosexual contact. One patient had cirrhosis with portal hypertension and one had raised alkaline phosphatase as the only abnormality on liver function tests. CONCLUSIONS: Because HCV is more infectious by the parenteral route, the incidence of coinfection is lower in our population possibly due to the fact that majority of our HIV positive patients acquire infection through heterosexual contact. PMID- 10695421 TI - A case of pancreatic hydatid disease. PMID- 10695422 TI - Hereditary pancreatitis. AB - Hereditary pancreatitis is a rare cause of chronic pancreatitis and has been mainly reported in western literature. It is inherited in an autosomal dominant manner. In this report, we present our data on two affected members in a family from Punjab, North India. PMID- 10695423 TI - Gastric adenocarcinoma--does the extent of lymphadenectomy matter? PMID- 10695424 TI - Upper gastrointestinal diseases in Saudi Arabian children. AB - In contrast to the experience in the adults, there are limited data concerning the efficacy and safety of upper gastrointestinal endoscopy (UGIE) in paediatric patients. The information on this procedure is very scanty from non-western countries. We analysed 72 children evaluated in Gizan, Saudi Arabia, an area of high endemic hepatitis B and chronic liver disease. The indications comprised abdominal pain (49%), UGI bleeding (24%) and evaluation of suspected portal hypertension. No abnormality was detected in 33 (46%). Mucosal inflammatory lesions (oesophagitis, gastritis and duodenitis) are the commonest abnormal lesions, occurring in 24 (33%). Duodenal ulcer (4 cases) and gastric ulcer (1 case) were relatively few. No case of malignancy was found. Sclerotherapy for variceal bleeding was effective in 4 patients. Helicobacter pylori was detected in 12 of 23 patients and associated with histologically identified gastritis in the majority of these cases. It is concluded that paediatric UGIE is safe and useful in the diagnosis and therapeutic intervention for UGI diseases in children. Our findings provide additional information on the pattern of diseases among Saudi Arabian children. PMID- 10695425 TI - Extended survival of carcinoma head of pancreas following palliative treatment. AB - A 48 year old woman presented with obstructive jaundice 10 years back. Upper gastrointestinal endoscopy revealed a growth infiltrating the ampulla of Vater, which was confirmed to be adenocarcinoma on cytology. At laparotomy, a large nodular growth was seen in the head of pancreas. Surgical resection could not be done because of encasement of superior mesenteric vessels, hence a cholecystojejunostomy was performed. The patient remained asymptomatic for 9 years, when she developed cholangitis. Duodenoscopy at this stage revealed an ulcerated growth at the ampulla and biopsy from the growth confirmed a well differentiated adenocarcinoma. A straight flap 10 F stent was placed in the common bile duct. Thereafter the patient has remained asymptomatic for more than a year. PMID- 10695426 TI - Gangrenous sigmoid volvulus in a pregnant woman. AB - A rare case of gangrenous sigmoid volvulus in a pregnant woman causing intestinal obstruction is reported. The patient had intrauterine foetal death. Laparatomy for resection of sigmoid colon and hysterotomy for removal of dead foetus was carried out. Terminal iliac colostomy with closure of rectal stump was done in the first surgery. The patient underwent colorectal anastomosis 2 months after the first operation and recovered uneventfully. PMID- 10695427 TI - Giant non-parasitic hepatic cyst with biliary communication. PMID- 10695428 TI - Treatment of substance misuse in the new century. PMID- 10695429 TI - Congress and the Pain Relief Promotion Act. PMID- 10695430 TI - Conflict, what conflict? PMID- 10695431 TI - Functional foods: health boon or quackery? PMID- 10695432 TI - More on herpes zoster lesions. PMID- 10695433 TI - Should doctors disclose mistakes? PMID- 10695434 TI - Toxicology and pathology of deaths related to methadone: retrospective review. AB - OBJECTIVES: To clarify the mechanisms and risk factors of methadone toxicity and to describe the findings of deaths related to methadone use Design Retrospective review of case notes in the records of the San Francisco Medical Examiner comparing the findings in cases where methadone was deemed the cause of death with findings in decedents where methadone was an incidental finding, and with 50 age-matched, disease and drug free, trauma victims. RESULTS: 38 cases out of the 3317 processed by our office during 1997-1998 were identified in which methadone had been detected. Cases were mostly male 28/38 (74%) and white, 28/38 (74%). In 17 of 38 cases death was deemed to have been caused by methadone toxicity. For the group the mean blood methadone concentration for all 38 patients, was 957 ng/ml SD = .681, SE = .14). The mean blood concentration of the main methadone metabolite (EDDP) was 253 ng/ml, SD = 529 ng/ml, SE = .089. The mean ratio of methadone in the blood to EDDP in the blood was 13.6:1 Values were not significantly different between cases in which methadone toxicity was the cause of death and in those in which it was an incidental finding. Cocaine, or the cocaine metabolite benzoylecgonine, was detected in the blood or urine of 16/38 cases (42%); morphine in one-third (13/38) and methamphetamine in only one. Pulmonary edema was evident in all cases, coronary artery disease in 9/38 (24%) and cirrhosis in 7/38 (18%) of the methadone users. Necrotizing fasciitis was the cause of death in 4 of the 38 methadone users (11%). Nationally, a sizeable percent of methadone deaths are from drugs diverted from treatment programs. CONCLUSIONS: The presence of methadone is often an incidental finding during postmortem examination which is unrelated to the cause of death. Postmortem measurements of methadone or its metabolite, or both, cannot be used in isolation to identify which deaths are associated with methadone toxicity. PMID- 10695435 TI - Is methadone a miracle cure or an alternative evil? PMID- 10695436 TI - Risk of hepatitis B infection among young injection drug users in San Francisco: opportunities for intervention. AB - OBJECTIVE: To compare the demographic characteristics and risk behaviors for hepatitis B infection among injection drug users younger than 30 years with those aged 30 or older and to evaluate participants' knowledge, attitudes, and experiences of infection, screening, and vaccination against hepatitis B virus. DESIGN: A systematic sample of injection drug users not currently in a treatment program were recruited and interviewed at needle exchange programs and community sites. PARTICIPANTS: 135 injection drug users younger than 30 years and 96 injection drug users aged 30 or older. RESULTS: Injection drug users younger than 30 were twice as likely as drug users aged 30 or older to report having shared needles in the past 30 days (36/135 [27%] vs 12/96 [13%]). Injection drug users younger than 30 were also twice as likely to report having had more than two sexual partners in the past 6 months (80/135 [59%] vs 29/96 [30%]). Although 88 of 135 (68%) young injection drug users reported having had contact with medical providers within the past 6 months only 13 of 135 (10%) had completed the hepatitis B vaccine series and only 16 of (13%) perceived themselves as being at high risk of becoming infected with the virus. CONCLUSION: Few young injection drug users have been immunized even though they have more frequent contact with medical providers and are at a higher risk for new hepatitis B infection than older drug users. Clinicians caring for young injection drug users and others at high risk of infection should provide education, screening, and vaccination to reduce an important source of hepatitis B infection. PMID- 10695437 TI - Vaccination could improve overall health in a high risk population. PMID- 10695438 TI - Income inequalities and health disparities. PMID- 10695439 TI - Slow-release morphine was not more effective than methadone in reducing neonatal abstinence syndrome. PMID- 10695440 TI - Brief physician advice reduced drinking in older adults. PMID- 10695441 TI - Exercise training for patients with chronic heart failure reduced mortality and cardiac events and improved quality of life. PMID- 10695442 TI - Cost-effective evaluation of acute viral hepatitis. PMID- 10695443 TI - All patients with acute hepatitis must be observed until the acute liver injury resolves. PMID- 10695445 TI - Does "white coat hypertension" increase the risk for any adverse outcome from hypertension? PMID- 10695444 TI - Use of methadone. PMID- 10695446 TI - An elderly woman with a warm, painful finger. PMID- 10695447 TI - Screening patients for alcohol, tobacco, and other drug misuse: the role of brief interventions. PMID- 10695448 TI - Controversy erupts over reuse of "single use" medical devices. PMID- 10695450 TI - Big health insurer gives physicians final say in treatment. PMID- 10695449 TI - Health situation in former communist bloc is dire, says Unicef. PMID- 10695452 TI - A declaration of interdependence. PMID- 10695451 TI - Addiction is a treatable disease, not a moral failing. PMID- 10695453 TI - Catheter ablation of chronic atrial fibrillation targeting the reinitiating triggers. AB - INTRODUCTION: We assessed the mode of reinitiation of atrial fibrillation (AF) after cardioversion and the efficacy of ablating these foci of reinitiation in patients with chronic AF. METHODS AND RESULTS: Fifteen patients, 7 with structural heart disease, underwent mapping and catheter ablation of drug resistant AF documented to be persistent for 5 +/- 4 months. In all patients, cardioversion was followed by documentation of P on T atrial ectopy and early recurrence, which allowed mapping of the reinitiating trigger or the source of ectopy. Radiofrequency (RF) ablation was performed at pulmonary vein (PV) ostia using a target temperature of 50 degrees C and a power limit of 30 to 40 W, with the endpoint being interruption of all local muscle conduction. A total of 32 arrhythmogenic PVs and 2 atrial foci (left septum and left appendage) were identified: 1, 2, and 3 or 4 PVs in 5, 3, and 6 patients. RF applications at the ostial perimeter resulted in progressively increasing delay, followed by abolition of PV potentials in 8, but potentials persisted in 6. A single ablation session was performed in 7 patients and 8 underwent two or three sessions because of recurrence of AF; ablation was directed at the same source due to recovery of local PV potential or at a different PV. No PV stenosis was noted either acutely or at repeated follow-up angiograms. Nine patients (60%) were in stable sinus rhythm without antiarrhythmic drugs at follow-up of 11 +/- 8 months. Anticoagulants were interrupted in 7 patients. CONCLUSION: PVs are the dominant triggers reinitiating chronic AF in this patient population. Elimination of PV potentials by ostial RF applications results in stable sinus rhythm in 60%. A larger group and longer follow-up are needed to investigate further the role of trigger ablation in curative therapy for chronic AF. PMID- 10695454 TI - Mapping and radiofrequency catheter ablation of the three types of sustained monomorphic ventricular tachycardia in nonischemic heart disease. AB - INTRODUCTION: Sustained monomorphic ventricular tachycardia (VT) associated with nonischemic cardiomyopathy (CMP) is uncommon. Optimal approaches to catheter mapping and ablation are not well characterized, but they are likely to depend on the VT mechanism. The purpose of this study was to evaluate the mechanisms of sustained monomorphic VT encountered in nonischemic CMP and to assess the feasibility, safety, and efficacy of catheter radiofrequency ablation for treatment. METHODS AND RESULTS: Twenty-six consecutive patients with nonischemic CMP referred for management of recurrent VT were studied. In 16 (62%) patients, VT was related to a region of abnormal electrograms consistent with scar and the response to pacing suggested a reentrant mechanism. In 5 (19%) patients, VT was due to bundle branch or interfascicular reentry. In 7 (27%) patients, the VT mechanism was focal automaticity, 4 of whom had evidence of tachycardia-induced CMP. After catheter ablation targeting parts of reentrant circuits, VT was not inducible in 8 (53%) of 15 patients with scar-related reentry, was modified in 5 (33%) patients, and still was inducible in 2 (13%) patients. Ablation was successful in 5 of 5 patients with bundle branch reentry and in 6 of 7 patients with a focal automaticity mechanism. Overall, catheter ablation abolished clinical recurrence of VT in 20 (77%) of 26 patients during a follow-up of 15 +/- 12 months. CONCLUSION: Three different mechanisms of VT are encountered in patients with nonischemic CMP. The mapping and ablation approach varies with the type of VT. In this selected population, the overall efficacy was 77%. PMID- 10695455 TI - Monomorphic ventricular tachycardia in nonischemic disease: what have we learned? PMID- 10695456 TI - Sensing lead failure in implantable defibrillators: a comparison of two commonly used leads. AB - INTRODUCTION: Despite major technological advances, structural problems in implantable cardioverter defibrillator (ICD) endocardial sensing leads remain a significant problem. There are two types of ICD sensing leads: (1) dedicated bipolar leads and (2) integrated lead systems that include defibrillation coils. The long-term performance of these two lead systems has not been directly compared. METHODS AND RESULTS: We prospectively examined the incidence of lead failure manifested by inappropriate arrhythmia detection in 247 consecutive patients undergoing abdominal ICD implant at a single center between 1991 and 1995. A total of 107 patients received BT-10 (dedicated bipolar) leads and 140 patients received Endotak (integrated bipolar) leads. Over a mean follow-up of 860 +/- 442 days, there were 19 (17.8%) lead failures with the BT-10 lead (261 to 1,505 days postimplant) compared with only 6 (4.3%; P < 0.01) with the Endotak lead (410 to 1,211 days postimplant). Lead failure was due to an insulation defect in all cases, with the problem occurring in the proximal lead (within the pulse generator pocket) in all but one case. Lead survival was significantly better with the Endotak lead (P = 0.015, risk ratio = 3.0, 95% confidence intervals 1.2 to 7.6). CONCLUSION: Late lead failure due to insulation defects in BT-10 sensing leads (causing inappropriate ICD activation) is a relatively common and progressive phenomenon, with difficulties becoming apparent as long as 4 years after implant. This problem is a likely cause of inappropriate shocks in patients with BT-10 leads. Implantation of a new sensing lead should be considered at the time of elective pulse generator replacement, even in the absence of demonstrable oversensing. PMID- 10695457 TI - Optimization of transvenous coil position for active can defibrillation thresholds. AB - INTRODUCTION: Lead systems that include an active pectoral pulse generator are now standard for initial defibrillator implantations. However, the optimal transvenous lead system and coil location for such active can configurations are unknown. The purpose of this study was to evaluate the benefit and optimal position of a superior vena cava (SVC) coil on defibrillation thresholds with an active left pectoral pulse generator and right ventricular coil. METHODS AND RESULTS: This prospective, randomized study was performed on 27 patients. Each subject was evaluated with three lead configurations, with the order of testing randomized. Biphasic shocks were delivered between the right ventricular coil and an active can alone (unipolar), or an active can in common with the proximal coil positioned either at the right atrial/SVC junction (low SVC) or in the left subclavian vein (high SVC). Stored energies at defibrillation threshold were higher for the single-coil, unipolar configuration (11.2 +/- 6.6 J) than for the high (8.9 +/- 4.2 J) or low (8.5 +/- 4.2 J) SVC configurations (P < 0.01). Moreover, 96% of subjects had low (< or = 15 J) thresholds with the SVC coil in either position compared with 81% for the single-coil configuration. Shock impedance (P < 0.001) was increased with the unipolar configuration, whereas peak current was reduced (P < 0.001). CONCLUSION: The addition of a proximal transvenous coil to an active can unipolar lead configuration reduces defibrillation energy requirements. The position of this coil has no significant effect on defibrillation thresholds. PMID- 10695458 TI - Electrophysiologic characteristics of the atrium in sinus node dysfunction: atrial refractoriness and conduction. AB - INTRODUCTION: Clinical electrophysiology (EP) has focused attention on the EP properties of atrial muscle in patients with atrial fibrillation (AF). Patients with sinus node dysfunction (SND) sometimes are included in these studies, but the characteristics of these patients with SND alone appear less well investigated. METHODS AND RESULTS: We reviewed EP data of 46 patients (mean age 70 +/- 8 years) with SND, who underwent EP study for evaluation of the atrial substrate. In 16 patients, a history of paroxysmal AF was documented, but not in the remaining 30 patients who had SND alone. We considered as control a group of 25 subjects (mean age 63 +/- 14 years), who were referred to our EP laboratory for unexplained syncope or AV conduction disturbances. Following pharmacologic washout and at a drive cycle of 600 msec, effective (ERP) and functional refractory periods (FRP), S1-A1 and S2-A2 latency, A1 and A2 width, latent vulnerability index (ERP/A2), and P wave duration on the surface ECG were measured. Intra-atrial conduction times were measured from the stimulus artifact by pacing the high right atrium (HRA), to the corresponding atriograms at the AV node (HRA-AVN), low lateral atrium (HRA-LLA), and low interatrial septum close to the coronary sinus ostium (HRA-CSO). Compared with the control group, SND patients did not show differences in ERP (238 +/- 26 msec vs 250 +/- 29 msec), FRP (274 +/- 25 msec vs 280 +/- 32 msec), S1-A1 (38 +/- 15 msec vs 33 +/- 11 msec) and S2-A2 latency (67 +/- 24 msec vs 63 +/- 25 msec), or HRA-AVN (81 +/- 24 msec vs 65 +/- 19 msec), HRA-LLA (36 +/- 30 msec vs 40 +/- 27 msec), and HRA-CSO (77 +/- 17 msec vs 80 +/- 15 msec) conduction times. In contrast, we observed strong differences in atriogram durations A1 (59 +/- 19 msec vs 39 +/- 13 msec; P < 0.001) and A2 (92 +/- 28 msec vs 57 +/- 18 msec; P < 0.001), as well as in the latent vulnerability index ERP/A2 (2.8 +/- 1.2 msec vs 4.8 +/- 1.7; P < 0.001). Also, the P wave was slightly longer (104 +/- 18 msec vs 94 +/- 45 msec; P < 0.05). No significant statistical difference in EP parameters was found between SND patients with or without documented AF. CONCLUSION: In patients with SND, atrial refractoriness appears similar to that of control subjects. The most important EP abnormality appears to be local conduction slowing disturbances, with prolonged basal and postextrastimuli atriograms, responsible for a lower vulnerability index. This could explain, at least in part, the tendency of patients with SND to develop AF during their natural history. Normality of atrial refractoriness, in contrast to atrial conduction disorders, might explain why atrial pacing shows a preventative effect on the development of AF and why antiarrhythmic drugs often are ineffective. PMID- 10695459 TI - Location of the lower turnaround point in typical AV nodal reentrant tachycardia: a quantitative model. AB - INTRODUCTION: Recent observations suggest that the circuit of AV nodal reentrant tachycardia (AVNRT) may extend down to the His bundle. The purpose of this study was to develop a quantitative model indicating the location of the lower turnaround point in AVNRT. METHODS AND RESULTS: Slow pathway modification was performed in 70 patients with typical AVNRT. During sinus rhythm, ventricular pacing was performed with the AVNRT cycle length. During AVNRT, the HinitAinit interval was measured from initial His to the initial atrial deflection recorded in the His-bundle lead. During ventricular pacing, the HendAinit interval was measured from end of the His to the beginning of the atrial deflection. It was hypothesized that x reflects conduction time from the lower turnaround point to Ainit, whereas y reflects conduction time from the lower turnaround point to Hinit. Anterograde conduction during AVNRT and retrograde conduction during ventricular pacing were assumed to be identical if there was 1:1 retrograde conduction at the AVNRT cycle length. The following formulas describe the relation of the measured parameters: x - y = HinitAinit; and x + y = HendAinit. Resolving both formulas yields the unknown x and y: y = (HendAinit - HinitAinit)/2, x = (HendAinit + HinitAinit)/2. These criteria were present in 52 of 70 patients. The mean cycle length of AVNRT was 355 +/- 42 msec, mean HinitAinit was 54 +/- 27 msec, and mean HendAinit was 60 +/- 29 msec. Accordingly, in 20 of 52 patients, the lower turnaround point was located within the His bundle (y = -15.4 +/- 16.1 msec), in 3 of 52 it was in the nodal-His junctional area (y = 0), and in 29 of 52 it was above the His bundle (y = +12.7 +/- 10.3 msec). The HinitAinit interval was significantly longer (66 +/- 32 msec vs 47 +/- 20 msec; P = 0.02) and the HendAinit interval was significantly shorter (45 +/- 30 msec vs 69 +/- 24 msec; P = 0.004) when the first group was compared with the others. CONCLUSION: In about 1 of 3 of patients with typical AVNRT, the lower turnaround point of the circuit is within the His bundle; in more than half of the patients it is above the His bundle. These data do not support the concept that all AVNRTs have an intranodal circuit, but are in accordance with the finding of longitudinal dissociation of the His bundle. PMID- 10695460 TI - Catheter ablation for hemodynamically unstable monomorphic ventricular tachycardia. AB - INTRODUCTION: Hemodynamic collapse precludes extensive catheter mapping to identify focal target regions in many patients with ventricular tachycardia (VT) associated with heart disease. This study tested the feasibility of catheter ablation of poorly tolerated VTs by targeting a region identified during sinus rhythm. METHODS AND RESULTS: Ablation was attempted in five patients, ages 44 to 59 years, with left ventricular ejection fractions of 0.15 to 0.20 and poorly tolerated VT causing multiple implantable defibrillator therapies (6 to 30 episodes/month). VT was due to prior infarction in three patients and nonischemic cardiomyopathy in two. Target regions were sought that met the following criteria: (1) evidence of slow conduction from fractionated sinus rhythm electrograms and stimulus-QRS delays during pace mapping, and (2) evidence that the region contains the reentrant circuit exit from pace mapping. In 4 of 5 patients, a target region was identified and radiofrequency lesions applied. Ablation abolished all recurrences of VT in 3 of 4 patients during follow-up of 14 to 22 months. There were no complications. CONCLUSION: Ablation of poorly tolerated VT is feasible in some patients by mapping during sinus rhythm and performing ablation over a region of identifiable scar that contains abnormal conduction and a presumptive VT exit. PMID- 10695461 TI - Ventricular tachycardias mimicking those arising from the right ventricular outflow tract. AB - INTRODUCTION: Ablation of ventricular tachycardia (VT) arising from the right ventricular outflow tract (RVOT) has proven highly successful, yet VTs with similar ECG features may originate outside the RVOT. METHODS AND RESULTS: We reviewed the clinical, echocardiographic, and ECG findings of 29 consecutive patients referred for ablation of monomorphic VT having a left bundle branch block pattern in lead V1 and tall monophasic R waves inferiorly. Nineteen patients (group A) had VTs ablated from the RVOT, and 10 patients (group B) had VTs that could not be ablated from the RVOT. The QRS morphology during VT or frequent ventricular premature complexes was the only variable that distinguished the two groups. During the target arrhythmia, ECGs of group B patients displayed earlier precordial transition zones (median V3 vs V5; P < 0.001), more rightward axes (90 +/- 4 vs 83 +/- 5; P = 0.002), taller R waves inferiorly (aVF: 1.9 +/- 1.0 vs 2.4 +/- 0.5; P = 0.020) and small R waves in lead V1 (10/10 vs 9/19; P = 0.011). Radiofrequency catheter ablation from the RVOT failed to eliminate VT in any group B patient, but ablation from the left ventricular outflow tract (LVOT) eliminated VT in 2 of 6 patients in whom left ventricular ablation was attempted. CONCLUSION: The absence of an R wave in lead V1 and a late precordial transition zone suggest an RVOT origin of VT, whereas an early precordial transition zone characterizes VTs that mimic an RVOT origin. The latter VTs occasionally can be ablated from the LVOT. Recognition of these ECG features may help the physician advise patients and direct one's approach to ablation. PMID- 10695462 TI - Augmentation of QRS wave amplitudes in the precordial leads during narrow QRS tachycardia. AB - INTRODUCTION: QRS morphology during narrow QRS supraventricular tachycardia in patients without ventricular preexcitation generally is considered the same as that seen during sinus rhythm. This study presents a new ECG observation that the QRS amplitude increased significantly in leads V2 through V5 during tachycardia. METHODS AND RESULTS: Using the same ECG machine and the same electrode patches applied to the same electrode positions, 12-lead ECGs during sinus rhythm and narrow QRS tachycardia were analyzed comparatively in 23 patients without ventricular preexcitation. Precordial QRS amplitudes were measured as the vertical distance from the peak of the R to the nadir of the S wave. The amplitudes also were measured during atrial rapid pacing and extrastimulation. Furthermore, ventricular excitation during sinus rhythm and tachycardia was studied using body surface mapping. Body surface distributions of QRS potentials and ventricular activation time (VAT) were displayed as maps. Gross area of QRS (AQRS, equivalent to the QRS amplitude) was compared during sinus rhythm versus tachycardia. During tachycardia, QRS amplitude significantly increased in leads V2 through V5, without any noticeable change in the transitional zone or QRS wave duration. Increase of QRS amplitude also was noted during atrial rapid pacing and extrastimulation. Gross AQRS values during tachycardia significantly increased in the left parasternal area, whereas QRS isopotential and VAT isochronal maps were similar during sinus rhythm and tachycardia, suggesting a minimal role of conduction delay in the increase of QRS amplitude. CONCLUSION: QRS wave amplitude significantly increased in leads V2 through V5 during narrow QRS tachycardia compared with QRS waves in sinus rhythm. Increase of QRS amplitude seemed unlikely due to a conduction delay within the ventricular myocardium. PMID- 10695463 TI - Effect of heart rate on QRS voltage: a simple relation that escaped notice. PMID- 10695464 TI - Transvenous parasympathetic nerve stimulation in the inferior vena cava and atrioventricular conduction. AB - INTRODUCTION: In previous reports, we demonstrated a technique for parasympathetic nerve stimulation (PNS) within the superior vena cava, pulmonary artery, and coronary sinus to control rapid ventricular rates during atrial fibrillation (AF). In this report, we describe another vascular site, the inferior vena cava (IVC), at which negative dromotropic effects during AF could consistently be obtained. Moreover, stimulation at this site also induced dual AV nodal electrophysiology. METHODS AND RESULTS: PNS was performed in ten dogs using rectangular stimuli (0.1 msec/20 Hz) delivered through a catheter with an expandable electrode basket at its tip. Within 3 minutes and without using fluoroscopy, the catheter was positioned at an effective PNS site in the IVC at the junction of the right atrium. AF was induced and maintained by rapid atrial pacing. During stepwise increase of the PNS voltage from 2 to 34 V, a graded response of ventricular rate slowing during AF was observed (266 +/- 79 msec without PNS vs 1,539 +/- 2,460 msec with PNS at 34 V; P = 0.005 by analysis of variance), which was abolished by atropine and blunted by hexamethonium. In three animals, PNS was performed during sinus rhythm. Dual AV nodal electrophysiology was present in 1 of 3 dogs in control, whereas with PNS, dual AV nodal electrophysiology was observed in all three dogs. PNS did not significantly change sinus rate or arterial blood pressure during ventricular pacing. CONCLUSION: Stable and consistent transvenous electrical stimulation of parasympathetic nerves innervating the AV node can be achieved in the IVC, a transvenous site that is rapidly and readily accessible. The proposed catheter approach for PNS can be used to control ventricular rate during AF in this animal model. PMID- 10695465 TI - Ionic basis for action potential prolongation by phenylephrine in canine epicardial myocytes. AB - INTRODUCTION: In canine ventricle, alpha-adrenergic agonists prolong action potential duration (APD) without any effect on the action potential notch, suggesting that, in this species, the effect on repolarization might be independent of inhibition of I(to). The present study investigated the action of the alpha-adrenergic agonist phenylephrine on the action potential and the repolarizing currents I(to) and I(K) in isolated canine epicardial myocytes. METHODS AND RESULTS: Isolated cells from canine epicardial tissue, and Purkinje fibers, were studied with the whole cell, voltage clamp method. Phenylephrine 0.1 microM increased APD by 13% +/- 4% at 90% repolarization without affecting the notch or amplitude. Under voltage clamp, concentrations of phenylephrine as high as 10 microM had no effect on I(to) in canine epicardial myocytes. However, I(to) of isolated canine Purkinje myocytes was reduced to 69% +/- 7% of control by 1 microM phenylephrine. Further studies in canine epicardial myocytes revealed an action of phenylephrine to inhibit I(K), and in particular I(Ks). Using a voltage protocol that included a two-step repolarization to separate I(Ks) and I(Kr) tail components, the largely I(Kr) component was not significantly affected by 1 microM phenylephrine, whereas the largely I(Ks) component was reduced to 81% +/- 5% of control value. CONCLUSION: Alpha-adrenergic prolongation of repolarization in canine epicardium does not result from inhibition of I(to). Rather, it appears that reduction of I(Ks) contributes to the action of phenylephrine. The unresponsiveness of epicardial I(to) is not a general characteristic of the canine heart, because Purkinje myocyte I(to) was inhibited, suggesting regional differences in the molecular basis of I(to) and/or alpha-adrenergic signaling in the canine heart. PMID- 10695466 TI - Effect of atrial radiofrequency ablation designed to cure atrial fibrillation on atrial mechanical function. AB - INTRODUCTION: The effects of linear radiofrequency lesions in the atria for cure of atrial fibrillation on atrial contraction have not previously been quantified. METHODS AND RESULTS: Atrial function was measured before and 30 +/- 24 days after a biatrial ablation procedure designed to cure atrial fibrillation in eight dogs and after a sham procedure in three dogs. Atrial mechanical function was assessed using Doppler diastolic blood flow velocities, atrial systolic pressure wave amplitude, and assessment of atrial contribution to cardiac output estimated by comparison of AV sequential pacing to ventricular pacing at the same heart rate. The mitral Doppler A/E velocity ratio was 1.03 +/- 0.45 before and 0.72 +/- 0.43 after ablation (P = 0.048). The tricuspid A/E ratio was 0.88 +/- 0.17 before and 0.71 +/- 0.12 after ablation (P = 0.04). The estimated atrial contribution to cardiac output was 18% +/- 9% before and 5% +/- 4% after ablation (P < 0.01). The left atrial systolic pressure wave amplitude was 2.8 +/- 1.5 mmHg before and 1.7 +/- 1.0 mmHg after ablation (P = 0.1). These changes were not observed in control dogs. Lesions covered 25% +/- 6% of the atrial endocardial surface. CONCLUSION: Multiple linear radiofrequency lesions in the atria designed to cure atrial fibrillation may impair atrial contractility. Reduced atrial function is partly due to loss of atrial myocardial mass, but regional delays in atrial activation and splinting of the atria by scarring also may contribute. PMID- 10695467 TI - How constant anatomically is the tendon of Todaro as a marker for the triangle of Koch? AB - INTRODUCTION: Although well recognized by anatomists as a border of the triangle of Koch demarcating the location of the AV node, the tendon of Todaro is not visible in the operating room or in the catheterization laboratory. Instead, clinicians use as surrogate a projected line between the eustachian valve and the central fibrous body. The constancy of the tendon of Todaro within this border remains to be determined. MATERIALS AND RESULTS: We reexamined serial histologic sections from 25 adults and 50 infants and gross dissections in four normal hearts. The tendon of Todaro was identified in all cases and traced to the central fibrous body in all but one case. It tended to be thicker in the hearts of infants cases (0.2 to 0.8 mm vs 0.1 to 0.6 mm). The tendon and the hinge-line of the septal leaflet of the tricuspid valve were consistent as landmarks for location of the compact AV node in all the cases studied by histology. Gross dissections traced the tendon to the free edge of the eustachian valve. CONCLUSION: The tendon of Todaro is present in hearts obtained from both adults and infants. It, or its surrogate, is a reliable border for the triangle of Koch and serves as a landmark to location of the atrial components of the AV conduction axis. PMID- 10695468 TI - Right atrial flutter isthmus revisited: normal anatomy favors nonuniform anisotropic conduction. AB - INTRODUCTION: The "flutter isthmus," the part of the lower right atrium between the eustachian valve and the tricuspid annulus inferior to the coronary sinus os, is considered the crucial zone for conduction delay necessary for the genesis of atrial flutter. However, the underlying mechanism remains unclear. METHODS AND RESULTS: We studied the "flutter isthmus" in 50 hearts obtained at autopsy from patients without atrial tachyarrhythmias. The muscular trabecular arrangement was dissected carefully by peeling off the endocardium. Documentation of the trabecular arrangement focused, in particular, on the question of whether there was a uniform pattern of well-aligned muscle trabeculae or a nonuniform architecture. It appeared that a nonuniform trabecular pattern prevailed (37/50 [74%]). In these hearts, the muscular arrangement showed abundant cross-overs and interlacing trabeculae, particularly in the zone immediately inferior to the coronary sinus os. Connections also occurred along the inferior rim of the os. CONCLUSION: The normal anatomy of the lower right atrium favors nonuniform muscular trabeculation, with interlacing bundles and a multitude of cross-overs. The potential for conduction delay is present in the vast majority of normal hearts. This raises the question as to what has changed in the hearts of patients with atrial flutter such that the potential for conduction delay and reentry has become effective. PMID- 10695469 TI - Ventricular fibrillation in a patient with prominent J (Osborn) waves and ST segment elevation in the inferior electrocardiographic leads: a Brugada syndrome variant? AB - Recurrent ventricular fibrillation was observed in a 29-year-old Vietnamese man who did not exhibit structural heart disease. The patient's ECG showed prominent J (Osborn) waves and ST segment elevation in the inferior leads that were not associated with hypothermia, serum electrolyte disturbance, or myocardial ischemia. Rate-dependent change in the amplitude of J waves and ST segment elevation also were observed. An implantable cardioverter defibrillator (ICD) was implanted. Adjunctive treatment with amiodarone reduced J wave amplitude, preventing ventricular fibrillation and ICD shocks. Prominent J waves and ST segment elevation in the inferior leads may serve as an important diagnostic sign to detect high-risk individuals with a history of unexplained syncope. ICD implantation plus amiodarone is the treatment of choice. PMID- 10695470 TI - An innovative application of anatomic electromagnetic voltage mapping in a patient with Ebstein's anomaly undergoing permanent pacemaker implantation. AB - A novel application of the Biosense CARTO System anatomic electromagnetic voltage mapping is presented, utilized as a guide for permanent pacemaker placement. The technique is illustrated in the successful implantation of an atrial lead in a patient with Ebstein's anomaly characterized by a severely dilated right atrium and extremely low-amplitude voltage signals, requiring a DDD pacemaker. Electromagnetic voltage mapping can be used in selected patients with structural heart disease to determine the optimal site for permanent pacemaker lead placement. PMID- 10695471 TI - Ablation of ventricular tachycardia by isolating the critical site in a patient with arrhythmogenic right ventricular cardiomyopathy. AB - We describe a patient with arrhythmogenic right ventricular cardiomyopathy in whom ventricular tachycardia (VT) was ablated by isolating a relatively large area of the critical site using catheter ablation. Endocardial mapping showed abnormal fragmented electrograms with delayed potential (DP) from an entire area of the aneurysm. Pace mappings from the aneurysm produced a QRS morphology identical to that of clinical VT. After catheter ablation was performed at the exit site of the VT critical area, programmed stimulation inside the aneurysm captured the DP but not the QRS complexes. These data suggest that VT can be ablated successfully by isolation of the critical area. PMID- 10695472 TI - Arrhythmias in heart failure: current concepts of mechanisms and therapy. AB - About one half of deaths in patients with heart failure are sudden, mostly due to ventricular tachycardia (VT) degenerating to ventricular fibrillation or immediate ventricular fibrillation. In severe heart failure, sudden cardiac death also may occur due to bradyarrhythmias. Other dysrhythmias complicating heart failure include atrial and ventricular extrasystoles, atrial fibrillation (AF), and sustained and nonsustained ventricular tachyarrhythmias. The exact mechanism of the increased vulnerability to arrhythmias is not known. Depending on the etiology of heart failure, different preconditions, including ischemia or structural alterations such as fibrosis or myocardial scarring, may be prominent. Reentrant mechanisms around scar tissue, afterdepolarizations, and triggered activity due to changes in calcium metabolism significantly contribute to arrhythmogenesis. Furthermore, alterations in potassium currents leading to action potential prolongation and an increase in dispersion of repolarization play a significant role. Treatment of arrhythmias is necessary either because patients are symptomatic or to reduce the risk for sudden cardiac death. The individual history, left ventricular function, electrophysiologic testing, and the signal-averaged ECG give useful information for identifying patients at risk for sudden cardiac death. The implantable cardioverter defibrillator (ICD) has evolved as a promising therapy for life-threatening arrhythmias. A potential role may exist for antiarrhythmic drugs, mainly amiodarone. There is growing evidence that patients with sustained VT or a history of resuscitation have the best outcome with ICD therapy regardless of the degree of heart failure. Many of these patients require additional antiarrhythmic therapy because of AF or nonsustained VTs that may activate the device. Catheter ablation or map-guided endocardial resection are additional options in selected patients but seldom represent the only therapeutic strategy. PMID- 10695473 TI - Gap junctions as active signaling molecules for synchronous cardiac function. PMID- 10695474 TI - Short ventriculoatrial intervals during orthodromic atrioventricular reciprocating tachycardia: what is the mechanism? PMID- 10695475 TI - Termination of ventricular tachycardia by a nonpropagated extrastimulus. PMID- 10695476 TI - Plaque rupture and plaque erosion. AB - There are multiple substrates for coronary thrombosis overlying an atherosclerotic plaque. The most common, plaque rupture, consists of an interruption of a thin fibrous cap overlying a lipid rich core. Plaque rupture is a result of macrophage infiltration and matrix degradation, is often seen in calcified plaques, and is highly associated with hypercholesterolemia. A less common substrate, plaque erosion, is not associated with elevated cholesterol and is the prime cause of coronary thrombosis in premenopausal women. The characteristic histologic features are abundant surface smooth muscle cells and proteoglycans, and a small or absent lipid rich core. The mechanisms of plaque erosion are unclear, and there are no consistent risk factors, although patients are often smokers. PMID- 10695477 TI - A role of plasminogen in atherosclerosis and restenosis models in mice. AB - In addition to its preeminent role in fibrinolysis, the plasminogen system is believed to play a key role in mediating cell migration. Leukocyte migration into the vessel wall is a key and early event in the development of the lesions of atherosclerosis and restenosis, pathologies which may be viewed as specific examples of vascular inflammatory responses. The development of mice in which the plasminogen gene has been inactivated affords an opportunity to test the contribution of plasminogen in leukocyte migration during in vivo. This article summarizes recent studies conducted in murine models of the inflammatory response, restenosis and atherosclerosis in which leukocyte migration, and in particular monocyte/macrophage migration, has been evaluated in plasminogen deficient mice. Recruitment of these cells through the vessel wall in inflammatory response models and into the vessel wall in restenosis and transplant atherosclerosis models is substantially blunted. These data implicate plasminogen in the migration of leukocytes in these murine models. With the numerous correlations between components and/or activation of the plasminogen system in restenosis and atherosclerosis, these results also support a role of plasminogen in the corresponding human pathologies. PMID- 10695478 TI - Coronary artery disease and fibrinolysis: from the blood to the vessel wall. AB - This review addresses a continuum of the role of derangements in the fibrinolytic system from the blood to the vessel wall in the pathogenesis of acute coronary events and the underlying vasculopathy. Pharmacologic modification of fibrinolysis has become a primary therapy for acute myocardial infarction caused by thrombotic occlusion of infarct-related coronary arteries. Patients with type 2 diabetes and other insulin resistant states exhibit impaired fibrinolysis in blood, implicated in the pathogenesis of macroangiopathy. An altered balance of activity of proteins involved in the fibrinolytic system within vessel walls (the proteo[fibrino]lytic system) has been recognized as well and appears likely to contribute to the acceleration of macroangiopathy. Accordingly, normalization of the proteo(fibrino)lytic system in blood and in vessel walls is a particularly attractive target for retardation of the progression of macrovascular disease associated with insulin resistant states. PMID- 10695479 TI - Plasminogen activator inhibitor type-1 (PAI-1) and its role in cardiovascular disease. AB - Cardiovascular disease is responsible for approximately 50% of total mortality in Europe, the USA and Japan. Established risk factors including smoking, hypercholesterolemia, and hypertension explain about half of the incidence of cardiovascular disease only. Reduced endogenous fibrinolytic activity secondary to increased plasma activity of plasminogen activator inhibitor type-1 (PAI-1) is now considered as a new cardiovascular risk factor. In this review, evidence is gathered for the notion that PAI-1 constitutes a predictor of cardiovascular disease and also contributes to the development of cardiovascular disease as a pathogenetic factor. The review will focus on experimental studies modulating PAI 1 activity and clinical studies addressing coronary heart disease, myocardial infarction, restenosis after coronary angioplasty, and graft occlusion after coronary artery bypass grafting. PMID- 10695480 TI - Structure and function of the urokinase receptor. AB - The binding of the urokinase plasminogen activator (uPA) to its receptor (uPAR) regulates cell adhesion, surface proteolysis, chemotaxis and cell extravasation in a number of experimental systems. Recent evidences have suggested that uPAR can by itself mediate chemotaxis of human monocytes and cause profound changes in cytoskeletal organization indicating that this receptor has the properties of a cell-surface regulated chemokine. Indeed, it is likely that upon binding to uPA, uPAR undergoes a conformational change that uncovers a new epitope located in the linker region between domain 1 and 2 of the receptor and is endowed with a potent chemotactic activity. This conformational change can be mimicked in vitro by enzymatic processing of a recombinant receptor. We have shown that chymotrypsin cleaves uPAR between domain 1 and 2 in an area that can be also cleaved by uPA at high efficiency and generate a receptor that can mediate monocytes migration independently of uPA binding. This mechanism is pertussis-toxin sensitive and involves activation of tyrosine kinases and cytoskeletal reorganization events in vitro. These studies indicate that in addition to its receptor function, upon binding to uPA, uPAR becomes a pleiotropic ligand for other still to be identified surface molecules. PMID- 10695481 TI - Control of smooth muscle cell proliferation--the role of the basement membrane. AB - In atherogenesis and in response to vessel injury, arterial smooth muscle cells (SMCs) are activated from a quiescent, differentiated state into an actively proliferating and synthetic phenotype which migrate into the intima where the cells participate in the formation of a fibrous plaque or intimal hyperplasia. The mechanisms involved in the control of SMC function are not clear and no preventive therapy against SMC activation is available. Interactions between SMCs and the extracellular matrix have been shown to influence SMC structure and function through integrin-mediated signaling processes. The SMC basement membrane is a specific form of extracellular matrix which seems to be crucial for the maintenance of SMC quiescence and the disruption of these interactions is part of cellular activation after atherogenic or traumatic stimuli. This concept of "negative growth control" may constitute a future target for the development of new strategies in the prevention of SMC activation in atherogenesis and restenosis formation. PMID- 10695482 TI - Gene regulation and arteriosclerosis: are developmental programs reactivated in vascular disease? AB - The molecular mechanisms regulating the development of vascular diseases such as atherosclerosis remain poorly understood at present. Similarities between genetic programs observed during the course of vascular disease with those observed during vascular development suggest that developmental processes are recapitulated in vascular disease. The earliest event in vascular development is the differentiation of endothelial cells from their mesodermally-derived hemangioblastic precursors. The receptor for vascular endothelial growth factor, KDR/flk-1, plays a critical role in these earliest stages of vascular development. During development and in the adult, expression of this receptor is restricted to vascular endothelial cells and their immediate precursors. We have therefore endeavored to determine the transcriptional events regulating KDR/flk-1 expression, with the hope of gaining insight into processes of vascular development that might also be important in vascular diseases of the adult. PMID- 10695483 TI - Oxidative stress and atherosclerosis: its relationship to growth factors, thrombus formation and therapeutic approaches. AB - The initiating event of atherogenesis is thought to be an injury to the vessel wall resulting in endothelial dysfunction. This is followed by key features of atherosclerotic plaque formation such as inflammatory responses, cell proliferation and remodeling of the vasculature, finally leading to vascular lesion formation, plaque rupture, thrombosis and tissue infarction. A causative relationship exists between these events and oxidative stress in the vessel wall. Besides leukocytes, vascular cells are a potent source of oxygen-derived free radicals. Oxidants exert mitogenic effects that are partially mediated through generation of growth factors. Mitogens, on the other hand, are potent stimulators of oxidant generation, indicating a putative self-perpetuating mechanism of atherogenesis. Oxidants influence the balance of the coagulation system towards platelet aggregation and thrombus formation. Therapeutic approaches by means of antioxidants are promising in both experimental and clinical designs. However, additional clinical trials are necessary to assess the role of antioxidants in cardiovascular disease. PMID- 10695484 TI - Soluble vascular cell adhesion molecule-1 (VCAM-1) as potential marker of atherosclerosis. AB - An increasing number of descriptive reports on soluble adhesion molecules and association with various diseases are published. Throughout these reports soluble adhesion molecules are identified as markers of inflammation. Since atherosclerosis demonstrates features of a chronic inflammatory disease, a potential association of soluble adhesion molecules with atherosclerosis has been postulated. However, conflicting results have been reported. One reason for this might be the differing definitions of atherosclerosis and patient groups. Besides the definition of atherosclerosis based on clinical symptoms, few reports use a direct quantification of atherosclerosis in their search for a marker of atherosclerosis. In those reports that quantify atherosclerosis, sVCAM-1 seems to be more specific for atherosclerosis than other markers. The serum level of sVCAM 1 appears to correlate with the extent of atherosclerosis and might allow for the detection of early stages of atherosclerosis. Large scale prospective studies will have to prove that sVCAM-1 can be used as a diagnostic tool for the detection of early stages of asymptomatic atherosclerosis and whether an early therapeutic intervention based on this approach is able to prevent progression and manifestation of the clinical sequelae of atherosclerosis. PMID- 10695485 TI - Angiogenesis in ischemic disease. AB - Angiogenic growth factors and their endothelial receptors function as major regulators of blood vessel formation. The VEGF/VEGFR and the Angiopoietin/Tie2 receptor systems represent key signal transduction pathways involved in the regulation of embryonic vascular development. Inactivation of any of the genes encoding these molecules results in defective vascular development and lethality between embryonic day 8.5 and 12.5. In addition, VEGF and its receptors are also critically involved in the regulation of pathological blood vessel growth in the adult during various angiogenesis-dependent diseases that are associated with tissue hypoxia, such as solid tumor growth and ischemic diseases. It is now well established that therapeutic angiogenesis can be achieved in animal models of hind limb and myocardial ischemia by exogenously adding VEGF and/or other angiogenic growth factors. Available clinical data from human trials also suggests that patients with severe cardiovascular diseases could potentially benefit from such therapies. However, much more work needs to be done to compare the potency of different angiogenic factors or the combination thereof, as well as the best way of delivery, either as recombinant proteins, as naked DNA or via adenoviral vectors. Nevertheless, the therapeutic efficacy of simply injecting naked plasmid DNA or proteins into ischemic tissue to deliver secreted angiogenic factors is an encouraging finding. Time will show whether the adverse side effects of therapeutic angiogenesis, mainly vascular permeability and edema formation, can be minimized and angiogenic factors can be used as an effective therapy in patients for the treatment of ischemic diseases such as arterial occlusive disease, myocardial infarction, and, eventually, also stroke. PMID- 10695486 TI - Unstable angina in 1998. AB - Unstable angina (UA) and non-Q-wave myocardial infarction (NQWMI) are acute coronary syndromes with repeated, severe ischemic events of short duration. These events are mainly due to a rapid decrease in coronary blood flow, and to a rapid, reversible reduction of the arterial lumen in localized areas. Episodes often are a mixture of thrombus formation due to platelet aggregation and localized spasm, leading to vasoconstriction. Due to the short interval (minutes) of ischemic events, usually no or minimal irreversible myocardial damage takes place. The main goal of treatment is to prevent progression of this unstable situation into a myocardial infarction. In the majority of cases, this is possible with adequate treatment of vasodilatory substances like nitrates, long-acting dihydropyridines like amlodipine and betablockers. In addition heparin and particular antiaggregatory drugs inhibiting platelet activation by blocking the GPIIb/IIIa receptor, the common pathway for platelet aggregation, are applied to prevent thrombus formation. This, in the majority of cases allows a passivation of the acute situation, leaving time to undertake possible further steps as coronary angiography, eventually followed by PTCA of the culprit lesion or, in advanced cases of CAD, by CABG with complete revascularization. PMID- 10695487 TI - Lipid lowering therapy and stabilization of atherosclerotic plaques. AB - Lipid-lowering therapy leads to a great reduction of cardiovascular complications, but has almost no effect on the degree of stenosis of coronary arteries. These and other studies have lead to a new paradigm of coronary artery disease, i. e. clinical prognosis is not only determined by the extent of a single stenosis, but mainly by the number and structure of atherosclerotic plaques. Rupture of an instable or vulnerable plaque, characterized by a large lipid-rich central core, inflammatory cells, and a thin fibrous cap, causes sudden thrombus formation and thereby acute coronary syndromes. There is accumulating evidence that cholesterol lowering can result in plaque stabilization and improvement of endothelial dysfunction. PMID- 10695488 TI - Acute myocardial infarction: selection of reperfusion strategies in the individual patient. AB - The selection of the reperfusion therapy (thrombolysis, PTCA, or no reperfusion) in acute myocardial infarction should depend on the risk-benefit ratio, the availability, and the costs. Thrombolysis is available everywhere and is the standard of reperfusion therapy, but carries the risk of intracranial haemorrhage. PTCA is more effective with less risk, but it is also more expensive and available only in specialised centres. In current clinical practise in Germany, the only important determinant for using PTCA as a reperfusion strategy is the availability of a catheter laboratory. Besides availability, the selection of reperfusion therapy should be made more on clinical relevance: (i) PTCA especially in patients with a large benefit, the presence of contraindications to thrombolysis, or in cardiogenic shock, and (ii) thrombolysis, if PTCA is not immediately available. PMID- 10695489 TI - Vectorcardiography: a tool for non-invasive detection of reperfusion and reocclusion? AB - In acute myocardial infarction, the perfusion status frequently fluctuates with rapid occurrences of coronary occlusion followed by myocardial ischemia. In patients with unstable angina, most episodes of ischemia are not accompanied by chest pain. In these patients it is important to be able to monitor the results of medical treatment non-invasively to establish the need for further intervention. It is not feasible to perform coronary angiography in all patients with acute myocardial infarction to evaluate patency of the infarct-related artery. Furthermore, even in a patent artery, no reflow may be present in the myocardial tissue. Angiography is therefore not the perfect golden standard to compare noninvasive ischemia monitoring with. Prognosis seems to be a better standard for comparison. This review indicates that vectorcardiography monitoring may identify myocardial reperfusion at an early stage and gives valuable prognostic information both in patients with unstable angina and acute myocardial infarction with low interobserver variability. PMID- 10695490 TI - Preinfarction angina and improved reperfusion of the infarct-related artery. AB - Preinfarction angina and early reperfusion of the infarct-related artery are major determinants of reduced infarct-size in patients with acute myocardial infarction. The beneficial effects of preinfarction angina on infarct size have been attributed to the development of collateral vessels and/or to post-ischemic myocardial protection. However, recently, a relation has been found between prodromal angina, faster coronary recanalization, and smaller infarcts in patients treated with rt-PA: those with preinfarction angina showed earlier reperfusion (p = 0.006) and a 50% reduction of CKMB-estimated infarct-size (p = 0.009) compared to patients without preinfarction angina. This intriguing observation is consistent with a subsequent observation of higher coronary recanalization rates following thrombolysis in patients with prodromal preinfarction angina compared to patients without antecedent angina. Recent findings in dogs show an enhanced spontaneous lysis of platelet-rich coronary thrombi with ischemic preconditioning, which is prevented by adenosine blockade, suggesting an antithrombotic effect of ischemic metabolites. Understanding the mechanisms responsible for earlier and enhanced coronary recanalization in patients with preinfarction angina may open the way to new reperfusion strategies. PMID- 10695491 TI - Ventricular remodeling after acute myocardial infarction. AB - The term ventricular remodeling has been coined to describe the geometrical changes in size and shape of the left ventricle occurring after large myocardial infarcts. We do not exactly know what initiates this process. Slipping of myofilaments following destruction of connective tissue--probably due to metalloproteinase activation--could be the initial event. As a consequence, wall stress is increased triggering deleterious adaptation processes, such as: - intracardiac angiotensin II generation; - cardiac endothelin formation and release; - pro-apoptotic signals for cardiomyocytes; - hypertrophic signals for fibroblasts and cardiomyocytes. This cascade of events is not only observed in the process of remodeling following myocardial infarction but is also operating during the progression of heart failure. Therapeutic principles therefore are similar in both conditions: - reduction of wall stress (pharmacological or mechanical unloading of the heart); - blockade of angiotensin II generation or of AT1-receptors (ACE-inhibitors or AT1 antagonists); - blockade of endothelin receptors (ET(A)-blockers); - blockade of adrenergic receptors (preferably beta1 adrenergic receptor blockers). Better understanding of the molecular mechanisms of the remodeling process already has fueled the search for new therapeutic interventions (such as endothelin receptor blockers, aldosterone antagonists and growth hormone application). Continuous research in this field may be especially rewarding if we will succeed in identifying the very first step in the cascade. PMID- 10695492 TI - Activation markers of coagulation and fibrinolysis: alterations and predictive value in acute coronary syndromes. AB - Several alterations of the coagulation, of the fibrinolysis and of inflammation are known in patients with acute coronary syndromes. To extent current knowledge of the pathophysiology and to optimize therapeutical strategies, the new molecular markers can be used in clinical studies. Furthermore, several studies were undertaken to assess the prognostic value of activation markers of these systems for patients with unstable angina pectoris and acute myocardial infarction with or without thrombolytic therapy. The majority of studies focussed on markers of thrombin activation, fibrinogen, fibrin degradation products and t PA and its main inhibitor PAI-1. While there are stimulating results from larger studies, the value for prognosis for the individual patient still is limited by the overlap of patients with good versus a poor outcome. PMID- 10695493 TI - Fibrinolytic response to venous occlusion compared to physical stress test in young patients with coronary artery disease. AB - INTRODUCTION: Venous occlusion (VO) and exercise stress (ES) are stimulators of the fibrinolytic system. Aim of this study was to answer which of both stimulation tests is more useful in patients with symptom-limited coronary artery disease (CAD) to evaluate possible defects in the fibrinolytic system. METHODS AND RESULTS: We investigated 20 patients (M/F = 15/5; mean age = 36.7 years) with angiographically proven CAD for their plasma levels of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-type-1 (PAI-1) at basal conditions as well as after VO and at maximal ES (standardised bicycle stress test) and compared the data to those obtained from 12 sex- and age-matched healthy controls (M/F = 9/3; mean age = 40.4 years). At basal conditions mean t PA activity and t-PA antigen plasma levels were within the normal range and comparable between the two study groups. After both VO and maximal ES, mean t-PA activity and t-PA antigen levels increased significantly more in the control group as compared to the CAD group. Mean PAI-1 activity plasma levels were significantly higher in the CAD group at basal conditions before VO (patients 7.0 +/- 3.1; controls 3.9 +/- 3.9; IU/ml; p = 0.025) as well as before ES (patients 8.1 +/- 3.5; controls 4.3 +/- 3.8; IU/ml; p = 0.009). PAI-1 activity plasma levels showed a significant decrease for patients and controls only after VO, while PAI-1 activity was not significantly decreased in both study groups at maximal ES. DISCUSSION: The significantly higher increase in mean plasma levels of t-PA activity and t-PA antigen after VO compared to ES in both groups might be explained by the fact that CAD induced symptoms in the patients during ES thus permitting only 80% of their age, sex, and body mass index related optimal work load. CONCLUSION: VO and ES are applicable triggers of the endogenous fibrinolytic system in healthy subjects and patients who are not limited in their physical exercise. Standardised VO appears to be superior to ES as stimulation test of the endogenous fibrinolytic system in patients with symptomatic CAD. PMID- 10695494 TI - Treatment of ischaemic stroke. AB - Stroke is a neurological emergency that requires rapid diagnostic workup and immediate treatment. Basic medical management such as maintenance of a high level of systemic blood pressure, provision of an optimal supply of oxygen, and correction of hyperthermia, hyperglycaemia and hypovolemia can contribute to a more favourable clinical outcome of stroke patients. Neuroprotective drugs have shown to be effective in reducing cerebral infarct size in several animal species but their clinical benefit in stroke patients remains to be proven. Neither heparin nor aspirin can improve neurological outcome or significantly reduce death or dependency several months after the ischaemic event, but aspirin is clearly effective in reducing early death or stroke recurrence within the first few weeks. Early anticoagulation should be used for patients at high risk for recurrent cardioembolic stroke and with carotid or vertebral artery dissection, although this recommendation is not based on the results of randomised controlled trials. PMID- 10695495 TI - Hemorrhagic transformation of cerebral infarction--possible mechanisms. AB - To analyse the risk/benefit of cerebral thrombolysis the role of hemorrhagic transformation, either as clinically silent hemorrhagic infarction or disastrous parenchymal hemorrhage, is crucial. Thrombolysis in acute ischemic stroke increases the risk of severe, life-threatening hemorrhagic complications by up to 10 times compared to controls. In this paper, previous proposed concepts for the development of intracerebral hemorrhage and hemorrhagic transformation are presented. The role of the cerebral microvasculature will be emphasized. In experimental focal cerebral ischemia a significant loss of basal lamina components of the cerebral microvessels has been demonstrated. This loss in vessel wall integrity is associated with the development of petechial hemorrhage. The mechanisms for this microvascular damage may include plasmin-generated laminin degradation, matrix metalloproteinases activation, transmigration of leukocytes through the vessel wall, and other processes. We propose that attenuation of the microvascular integrity loss with subsequent reduction in hemorrhage is theoretically possible 1) by an improvement in the definition of an individual time window of therapy (by means of imaging techniques), 2) by a biochemical quantification of the basal lamina damage to avoid dangerous interventions, and 3) by pharmacological strategies to protect the basal lamina during thrombolysis. PMID- 10695496 TI - Secondary prevention of stroke. AB - Stroke is a common disorder and a leading cause of disability and death. Ischaemia is a more common cause than haemorrhage and radiological imaging is required to accurately differentiate these. Some specific risk factors for stroke are non-modifiable--these include age, gender, racial and hereditary factors. Certain risk factors for ischaemic stroke can be identified and modification of these can be used for secondary prevention--examples include hypertension, heart diseases, atrial fibrillation, diabetes mellitus, dyslipidaemia, smoking, excessive alcohol consumption and carotid stenosis. Carotid endarterectomy is valuable in selected patients. In ischaemic stroke and transient ischaemic attack antithrombotic therapy is an option used in secondary prevention. In atrial fibrillation, warfarin should be used where possible in secondary prevention. When warfarin is contraindicated aspirin should be used. In other patients, an antiplatelet regime is appropriate--aspirin is commonly used and is the least expensive regime. Other antiplatelet agents such as dipyridamole, ticlopidine and clopidogrel may have a place. Younger patients with ischaemic stroke may have a thrombophilia state and should be appropriately investigated. PMID- 10695497 TI - Submassive and massive pulmonary embolism: a target for thrombolytic therapy? AB - Thrombolytic agents have been consistently demonstrated to dissolve pulmonary thrombi much more rapidly and effectively than heparin alone. Rapid resolution of pulmonary embolism (PE) is accompanied by a significant decrease in pulmonary artery pressure and an improvement in right ventricular function. However, it is no longer than 7 days until the findings of patients treated with heparin improve to a similar extent. Previous studies were not designed to determine whether this short-lasting difference in favor of thrombolysis can indeed affect the prognosis of patients with PE and thus justify the 1% (or even higher) risk of cerebral or fatal bleeding. Recently, two large registries demonstrated the importance of right ventricular dysfunction assessed by echocardiography as an independent predictor of mortality. Thrombolytic treatment was shown in one of these registries to be associated with a 50% reduction of death risk in clinically stable patients with right ventricular enlargement. It was thus possible to identify a group of patients with massive PE who are most likely to benefit from early thrombolysis. These findings now have to be confirmed by a prospective randomized trial which will compare thrombolysis with heparin alone in this high risk patient population, focusing on clinical end points such as overall and event-free survival in the acute phase of PE. PMID- 10695498 TI - Thrombolysis in arterial occlusion. AB - Intra-arterial thrombolytic therapy has replaced systemic intravenous infusion of thrombolytic agents as a treatment modality for arterial occlusion in the limbs. Several catheter-guided techniques and various infusion methods and schemes have been developed. At present there is no scientific proof of definite superiority of any agent in terms of efficacy or safety but clinical practice favours the use of urokinase or alteplase. Studies which compared thrombolysis to surgical intervention suggest that thrombolytic therapy is an appropriate initial management in patients with acute occlusion of a native leg artery or a bypass graft. Underlying causative lesions are treated in a second step by endovascular or open surgical techniques. Severe bleeding is the most feared complication: the risk of hemorrhagic stroke is 1-2%. PMID- 10695499 TI - Thrombolysis in newborns and infants. AB - This review analyses literature reports from 1970 to 1998 assessing the use of streptokinase (SK), urokinase (UK) or recombinant tissue-type plasminogen activator (rt-PA) for thrombolytic therapy in neonates and infants. From 1970 to 1998 182 infants were reported to have received SK (n = 54; 29.5%), UK (n = 41; 22.5%) or rt-PA (n = 87; 48%). During thrombolytic therapy no concomitant heparin administration or low dose heparin therapy (5 U/kg/h) were recorded. To perform reocclusion prophylactics heparin was reinitiated at the end of thrombolytic therapy usually in the recommended dosage of 20 U/ kg/h. The overall thrombolytic patency rate in neonates varied from 39% to 86%. Besides bleeding from local puncture sites or recent catheterisation sites (10.4%), pulmonary embolism was reported in 1.1% of the 182 infants. Major bleeding complications, i.e. pulmonary bleeding (0.6%), gastrointestinal bleeding (0.6%) or intraventricular haemorrhage (IVH 2.7%) are rarely reported side effects and only 2 thrombolysis related deaths due to haemorrhage were mentioned. Bleedings reported in the central nervous system (n = 4) mainly occurred in preterm infants (n = 3). In conclusion, data of this preliminary analysis suggest that there is no big difference (p = 0.09; chi2-test) in the efficacy rate between the 3 thrombolytic agents used in the first year of life. In each case an assessment must be made with respect to the relative benefit conferred by thrombolytic therapy in preventing organ or limb damage versus the potential side effects, costs and inconvenience for the childhood patient. Controlled prospective multicentre studies on thrombolytic therapy in neonates and infants are recommended to evaluate patency rates and adverse effects for the different thrombolytic agents used. PMID- 10695500 TI - TNK-tPA for acute myocardial infarction: the clinical experience. AB - Thrombolytic therapy has become the mainstay of treatment for acute transmural myocardial infarction. Present fibrinolytic regimens have a number of shortcomings, including the failure to induce early and sustained reperfusion in as many as 40-50% of the patients, and to prevent reocclusion in another 10-20% of the patients. The efforts for improving thrombolysis are focused on the development of new agents (fibrinolytics, anticoagulants, and antiplatelet agents). TNK-tPA is a triple combination mutant of wild-type tissue plasminogen activator that exhibits a longer plasma half-life, an enhanced fibrin specificity, and an increased resistance to the plasminogen activator inhibitor 1. This paper summarizes the results of clinical trials with TNK-tPA in acute myocardial infarction. PMID- 10695501 TI - Pharmacokinetics and pharmacodynamics of lanoteplase (n-PA). AB - In acute myocardial infarction rapid, complete, and sustained reperfusion of the infarct-related coronary artery is the most important therapeutic principle. Lanoteplase or n-PA, a third-generation plasminogen activator consisting of a deletion and point mutant of tissue-type plasminogen activator (t-PA), is a promising agent to approach this therapeutic goal. The molecule exhibits an increased plasma half-life allowing single-bolus administration. In this article, after characterizing the n-PA molecule, the currently available pharmacokinetic and pharmacodynamic data including the results of the InTIME study are reviewed. PMID- 10695502 TI - Oral anticoagulation after a first episode of venous thromboembolism: how long? How strong? AB - A number of studies have been published in the last years which shed light on the optimal intensity and the optimal duration of oral anticoagulation in patients with venous thrombosis. Based on these studies it is now generally recommended to treat patients with venous thromboembolism at an INR ranging from 2.0 to 3.0. The optimal duration of anticoagulation mainly depends on the nature of the thrombotic event. In patients with a temporary prothrombotic risk factor such as surgery, immobilization or trauma a relatively short duration of oral anticoagulation (3-6 months) is generally recommended. Patients with idiopathic venous thromboembolism require a considerably longer duration of anticoagulation (6 months at least). PMID- 10695503 TI - Bed rest in deep vein thrombosis and the incidence of scintigraphic pulmonary embolism. AB - In several countries of central Europe, patients with acute proximal deep vein thrombosis (DVT) are treated not only by anticoagulation and compression therapy but additionally by strict bed rest for 6-8 days. Until now the theoretical assumption that bed rest substantially reduces the incidence of pulmonary embolism has not been subjected to empirical verification. Patients with acute proximal DVT proven by ultrasonography were randomly assigned to strict bed rest for 8 days (treatment group) or to stay mobilised (control group). In both groups, basic treatment consisted in anticoagulation by subcutaneous low molecular weight heparin/vitamin-K-antagonist and compression therapy. The incidence of pulmonary embolism was assessed by serial ventilation/perfusion SPECT on day 1 and days 8-10. Of the 309 patients with proximal DVT considered for inclusion, 180 were excluded according to the study protocol, and 3 did not give informed consent. One hundred and twenty-six patients were randomly assigned to observe bed rest (n = 62) or to keep mobilised (n = 64). Four patients refused follow-up lung scan. A new lung perfusion defect was detected in 10/59 patients in the treatment group compared to 14/63 patients in the control group (one-sided p-value = 0.25; power 0.8). Bed rest as an additional measure in the treatment of DVT is not able to substantially reduce the incidence of scintigraphically detectable pulmonary embolism. The discomfort and costs associated with the prescription of bed rest in DVT are obviously inappropriate. PMID- 10695504 TI - Anticoagulation after artificial valve replacement with or without atrial fibrillation: how much is really needed? AB - Insertion of a mechanical artificial heart valve is an absolute indication for the use of lifelong oral anticoagulation, irrespective the presence or absence of atrial fibrillation. Depending on the type and position of the artificial valve the optimal International Normalized Ratio (INR) for these patients is between 2.5 and 4.5, although more prospective studies on the optimal range of oral anticoagulation are necessary. Addition of low dose antiplatelet therapy may further decrease the rate of thromboembolism, but also here more studies are needed. Patients with a bioprosthesis need only oral anticoagulation for the first three months following surgery in the absence of an indication for anticoagulation for other conditions. Atrial fibrillation is often seen in patients with artificial heart valves. Stroke prevention can be instituted by the mandatory oral anticoagulant therapy. The presence of atrial fibrillation and the use of oral anticoagulation prior to heart valve surgery support the indication for the insertion of a mechanical artificial heart valve. However, relative contraindications to oral anticoagulation and the presence of sinus rhythm may favor the use of a bioprosthesis. PMID- 10695505 TI - Platelet glycoprotein IIb/IIIa receptor blockade: lessening the risk of coronary interventions. AB - For the patient undergoing percutaneous coronary intervention, the administration of a platelet glycoprotein IIb/IIIa receptor blocker reduces the incidence of a periprocedural nonfatal myocardial infarction and the need for unplanned emergency stenting. In the diabetic patient undergoing intracoronary stenting, the use of a platelet glycoprotein IIb/IIIa receptor blocker appears to decrease the need for subsequent target vessel revascularization. There is considerable evidence in support of the use of glycoprotein IIb/IIIa receptor inhibitors in all categories of patients--"high-risk" patients, "low-risk" patients, and those undergoing primary angioplasty for acute myocardial infarction--and for the full armamentarium of percutaneous procedures (angioplasty, directional atherectomy, rotational atherectomy, and intracoronary stenting). PMID- 10695506 TI - Low-molecular weight heparins in venous and arterial thrombotic disease. AB - INTRODUCTION: Since the introduction of low-molecular-weight heparins (LMWHs) in the early 1980's, the use of these compounds has been extensively investigated as a substitute for unfractionated heparin (UFH) in patients with venous and arterial thrombotic diseases. LMWHs have several advantages as compared to UFH, such as the subcutaneous route of administration, the predictable anticoagulant response and the lack of the need for laboratory monitoring. The present systematic review evaluates randomised clinical trials which investigated the efficacy and safety of LMWH in the acute treatment of venous thromboembolism, myocardial infarction, unstable coronary syndromes and ischemic stroke. METHODS: A computerised and manual search was performed to identify all relevant clinical trials. All randomised studies, with an a priori defined study population, clinical outcome measurement and adequate follow-up, were reviewed by two independent assessors. Whenever possible a common effect estimate of the included studies was calculated. RESULTS: Thirteen studies in approximately 4000 patients with acute venous thromboembolism revealed an odds ratio for the 3-month recurrent thromboembolism rate and major bleeding complications during exposure of 0.77 (C.I. 0.57-1.04) and 0.61 (C.I. 0.39-0.95), respectively, in favour of LMWH as compared to UFH. In patients with acute myocardial infarction, one study suggested a reduction in the incidence of reinfarction and cardiac death in LMWH recipients compared to UFH, while a placebo-controlled study revealed no beneficial effect of LMWH on these outcomes. In six studies including over 7000 patients with acute unstable coronary syndromes, there was an odds ratio for recurrent angina, myocardial infarction, urgent revascularisation and major bleedings of 0.88 (C.I. 0.76-1.01), 0.84 (C.I. 0.69-1.01), 0.83 (C.I. 0.70-0.99), 1.09 (C.I. 0.70-1.70), respectively, in favour of LMWH compared to UFH. The three studies comparing LMWH treatment with placebo in approximately 1000 patients with acute ischemic stroke revealed an odds ratio for the 10-day recurrent stroke, death or disability after 3 months and major bleeding complications of 0.68 (C.I. 0.41-1.13), 0.94 (C.I. 0.78-1.15), 2.92 (C.I. 1.88-4.55), respectively. CONCLUSION: Fixed-dose subcutaneous LMWH appears to be a safe and effective alternative for dose-adjusted intravenous heparin in the treatment of patients with acute venous thrombotic disease as well as in patients with acute unstable coronary syndromes. The effectiveness of LMWH in patients with acute myocardial infarction remains unclear. There seems to be no beneficial effect of LMWH treatment as compared to placebo in patients with acute ischemic stroke, while the risk of major bleeding was clearly increased. PMID- 10695507 TI - Heparin-induced thrombocytopenia--pathogenesis and treatment. AB - Heparin-induced thrombocytopenia (HIT) is now recognized as the most frequent immune-mediated adverse drug reaction. During the last decade, fundamental aspects of the pathogenesis of HIT have been resolved. The understanding of some the mechanisms underlying the development of new, paradox thromboembolic complications in HIT led to the concept that thrombin generation plays a key-role in clinically manifest HIT. Consequently new therapeutic concepts imply the use of drugs with either indirect or direct anti-thrombin activity such as donaparoid sodium and the recombinant hirudin lepirudin. During the last years results of first prospective studies assessing various treatment regimens in HIT became available. Although data of randomized trials are still missing some treatment recommendations can already be drawn from these studies. This review summarizes key aspects of the pathogenesis of HIT and provides an overview of current treatment strategies. PMID- 10695508 TI - Ultrasound thrombolysis. AB - The possibility of lysing clots by the direct intravascular application of ultrasound was described first in 1974 in an animal experiment. However research on therapeutic ultrasound for thrombolysis gained momentum in the early 1980s and is now divided into several directions: 1) pharmacological thrombolysis supported by externally applied ultrasound; 2) pharmacological thrombolysis accelerated by intravascular ultrasound; 3) lysis of intravascular clots by intravascular ultrasound applied as singular treatment; 4) facilitating angioplasty by intravascular ultrasound which may smoothen the rigid surface of calcified arteries by lysing calcified structures out of sclerotic plaques. In acute myocardial infarction first evidence of efficacy in lysing clots by ultrasound was described in two small series of 15 patients in 1997. Furthermore, ultrasound thrombolysis seems to be efficient also in occluded saphenous vein grafts and in completely obstructed peripheral arteries, where the latest clinical experience gives very promising results. PMID- 10695509 TI - Primary stenting for acute myocardial infarction. AB - Primary stenting for acute MI has been shown to be an improvement over PTCA alone. As with primary PTCA however there is an obligate delay in restoration of TIMI flow due to the time necessary for mobilization of the cath lab team. It is possible that a hybrid approach using partial thrombolysis plus early IIB/IIIA inhibitor administration followed by urgent angiography and stenting of the culprit lesion will be the ideal approach. PMID- 10695510 TI - Prevention of restenosis after percutaneous coronary interventions: the medical approach. AB - Restenosis following successful percutaneous coronary revascularization continues to represent a major problem limiting the clinical efficacy of this procedure. The underlying mechanisms of restenosis are comprised of a combination of effects from vessel recoil, negative vascular remodeling, thrombus formation and neointimal hyperplasia. Indeed, there are important interactions among all of these mechanisms. For example, neointimal hyperplasia is stimulated by growth factors, which are released by local thrombi and the injured arterial segment itself, and act to enhance the expression of other growth-regulating proteins, in particular "second messengers", proto-oncogenes and other cell cycle controlling proteins. This results in an inflammatory and myofibroproliferative response, which may worsen vessel narrowing caused by recoil and result in the formation of a clinically significant restenotic lesion. A multitude of pharmacologic trials have been conducted in an attempt to prevent restenosis, but most have demonstrated little benefit. Studies in smaller numbers of patients have suggested a potential benefit for several classes of agents, including: 1) the antiproliferatives, angiopeptin, trapidil and tranilast; 2) selective elimination or alteration of proliferating cells; 3) enhancement of natural growth inhibitors; and 4) signal transduction blockade or inhibition of the gene expression for various growth-stimulating proteins. Finally, there have been advances in related areas, including development of antithrombotic catheters, novel polymers, and more efficient methods for transferring genes into the vessel wall. All of these offer the possibility of delivering agents (drugs, genes, or antisense oligonucleotides) locally at the site of intervention in a way that may optimize antiproliferative effects while minimizing systemic effects--ultimately leading to a more specific inhibition of the restenosis process. PMID- 10695511 TI - Stents for the treatment of aortic aneurysms. Review of devices, technique and results. AB - The endovascular treatment of abdominal aortic aneurysms has generated a great deal of interest since the early 1990s, and many different devices are currently available. The procedure of endovascular repair has been evaluated in many institutions and the different devices are compared. The first results were encouraging, but complications like endoleak, dislocation or thrombosis of the graft occurred. By the available devices the stent application is only promising, if the known exclusion criteria are strictly respected. Therefore a careful preinterventional assessment of the patient by different imaging modalities is necessary. As the available results up to now are preliminary and the durability of the devices has to be controlled, multicenter studies are required to improve the devices and observe their long-term success in the exclusion of abdominal aortic aneurysms. PMID- 10695512 TI - Treatment of carotid artery stenosis by elective stent placement instead of carotid endarterectomy in patients with severe coronary artery disease. AB - Patients with concomitant cardiac and cerebrovascular disease undergoing revascularization procedures are at high risk of both, cardiac and cerebrovascular complications. The purpose of our study was to evaluate the feasibility of prior elective carotid artery stenting as an alternative treatment procedure to carotid endarterectomy (CEA) in patients with concomitant coronary artery disease (CAD), who clearly needed coronary revascularization. We offered extracranial internal carotid stenting to 85 patients with 89 significant carotid stenoses. Out of these, 19 patients were symptomatic. The quantitative mean reduction in diameter was 77 +/- 11%. Stent implantation was successful in 88 lesions. Two disabling major and 3 reversible minor strokes occurred periprocedurally. Three patients showed asymptomatic restenosis and stent deformation was detected in 2 patients. Based on this experience, carotid stenting in high risk patients with severe coronary artery disease is feasible and safe and might be indicated as an alternative procedure for combined surgery. PMID- 10695513 TI - Identifying substance abusing delivering women: consequences for child maltreatment reports. AB - OBJECTIVE: The major objective was to determine how and the extent to which SB2669, which requires the identification of substance abusing delivering women, affected the number of children reported for abuse or neglect in several California counties. METHOD: A monthly time-series model from April 1988 to December 1995 was constructed. The idea underlying the model was that month-to month changes in the number of child maltreatment reports was a response to the presence of SB2669 in addition to various demographic, social, and economic factors. By separately estimating each county's number of reports, it was assumed that SB2669 did not necessarily affect each county's reports by the same amount, perhaps partially because of different counties' implementation strategies or general policies. Our sampling size consisted of seven high prevalence counties. RESULTS: The results suggest that the effects of SB2669 on the number of child maltreatment reports are mixed. On an aggregate level, all else constant, and at least for a few years after the passage of SB2669, SB2669 is associated with a decrease in child maltreatment reports in two of the participating counties. This decrease may be due to conscientious implementation of the legislation in these counties. This mixed finding is expected mainly because SB2669, although mandated, was never enforced. Moreover, from the process component of the study we learned that the implementation practices of this legislation vary substantially between and even within counties' hospitals. CONCLUSIONS/FUTURE DIRECTIONS: A decrease in maltreatment reports in the presence of SB2669 is not necessarily the most desirable outcome in the light of what we know about the relationship between substance abuse and child maltreatment. Law makers need to rethink the purpose of the law and provide the necessary language, tools and training to ensure that the goals of identifying substance abusing mothers and their families are met. Provisions also need to be made that somehow enforce this legislation. These provisions could lessen county-level and hospital-level variability in implementing the law. PMID- 10695514 TI - Maltreatment of children with disabilities: training needs for a collaborative response. AB - PROBLEM STATEMENT: There is a dearth of research on how to respond to children with disabilities who have been maltreated. The literature that does exist recommends a collaborative team approach, with each team member possessing a broad understanding of the special considerations of working with children with disabilities. The literature does not define current understanding levels of response team members in comparison to essential knowledge levels. METHOD: The current study used a needs assessment instrument tailored to each of three key groups: parents, educators, and investigators. Respondents were asked about their knowledge level, experience with, and training interests on maltreatment of children with disabilities. RESULTS: While respondents seemed to have a cursory awareness in some of the topic areas, their knowledge levels were not extensive in most of the survey areas. A majority of respondents were willing to attend training, and all three groups ranked the recognition of maltreatment of children with disabilities as a top training priority. CONCLUSIONS: It was concluded that these integral players in the response to maltreatment of children with disabilities are receptive to becoming more effective partners, by attending training to bridge the knowledge gaps they possess. The current study helps document the nature of those knowledge gaps and, thereby informs the development of training programs for building a more coordinated and informed response to maltreatment of children with disabilities. PMID- 10695515 TI - Enhancing law enforcement identification and investigation of child maltreatment. AB - OBJECTIVE: Data from two independent studies is presented, representing the investigators' ongoing work with faculty from a state criminal justice academy to analyze existing knowledge and skill levels among veteran law enforcement officers and recruits, as well as to enhance future training. METHOD: Through an anonymous questionnaire, the first of these studies examined officers' perceptions of maltreatment, including those factors that do and do not influence a determination of whether a particular act constitutes child maltreatment and assessments of whether particular acts constitute abuse or neglect. The second study also utilized an anonymous questionnaire to examine officers' knowledge of the developmental strengths and limitations of children relative to their ability to provide accurate information in suspected cases of child maltreatment. RESULTS: As hypothesized, several gaps both in law enforcement officers' knowledge of certain characteristics that can serve to denote a case of maltreatment and their knowledge of fundamental developmental issues and interview techniques that could assist them in the performance of their professional duties are identified. CONCLUSIONS: Suggestions for enhanced law enforcement training programs are presented and discussed. PMID- 10695516 TI - The effects of daily stressors on physical health in women with and without a childhood history of sexual abuse. AB - OBJECTIVE: The primary purpose of the present study was to examine the relationship between daily stressors and physical symptoms in college-age women with a childhood history of sexual abuse and women without a history of childhood sexual abuse. It was hypothesized that women with a history of childhood sexual abuse would be particularly susceptible to the effects of daily stressors on physical symptoms, and would show more covariation between daily stressors and physical symptoms, compared to women without a history of childhood sexual abuse. METHOD: Female college students (n = 491) were screened for histories of childhood (before age 15) and adulthood (after age 15) contact sexual abuse. Of these participants, 18 women with only a history of childhood sexual abuse were assigned to the SA group, and 27 women with no history of childhood or adulthood sexual abuse were assigned to the NA group. These women filled out self-report measures of daily hassles and physical symptoms for 28 consecutive days. RESULTS: During the 5 days preceding a highly stressful day, women in the SA group reported significantly more physical symptoms than during the 5 days preceding a day of low stress. For the NA group, there were no significant differences in reported physical symptoms between high- and low-stress days. CONCLUSIONS: The pattern of results for physical symptoms suggests that women with a history of childhood sexual abuse may be particularly susceptible to the effects of heightened daily stress, and may display this susceptibility in the report of physical symptoms. Possible explanations for these findings are discussed. PMID- 10695517 TI - Child characteristics which impact accuracy of recall and suggestibility in preschoolers: is age the best predictor? AB - OBJECTIVE: The purpose of the present study was to determine whether individual difference factors of metamemory, intelligence, and temperament can improve the ability to predict accuracy of recall and suggestibility in preschoolers. METHOD: Fifty-six children ranging in age from 43 months to 83 months (M = 61, SD = 9) were recruited from 13 child care centers in a rural southeastern town. Children participated in a "Circus Day" event conducted by two female undergraduate psychology students dressed as clowns. Approximately 10 days (M = 10; SD = 2) after the event, children were interviewed regarding their experiences. RESULTS AND CONCLUSIONS: Bivariate correlations and multiple regression analyses were performed in order to determine which factors were related and unique contributors to accuracy of recall and suggestibility. Of principal importance is the finding that child characteristics such as metamemory ability, intellectual functioning, and temperament may indeed be helpful in determining a child's capacity to accurately recall information in an interview, although for the most part age is the best predictor. Findings also underscore the importance of considering a child's SES and race when planning and conducting interviews with young children. Possible explanations for these findings as well as implications for future research and clinical application are discussed. PMID- 10695518 TI - Differentiating incest survivors who self-mutilate. AB - OBJECTIVE: This study was an exploratory analysis of the variables which differentiated incest survivors who self-mutilate from those who do not. METHOD: A sample of women incest survivors (N = 84) were divided into two groups based on the presence or absence of self-mutilation. Participants included both community and clinical populations. A packet consisting of a demographic questionnaire, Sexual Attitudes Survey, Diagnostic Inventory of Personality and Symptoms, Dissociative Events Scale and the Beck Depression Inventory was completed by each participant. RESULTS: Demographic, incest, and family of origin variables distinguished the self-mutilating women from those who did not. These include ethnicity and educational experiences; duration, frequency, and perpetrator characteristics regarding the incest; and multiple abuses, instability, birth order, and loss of mother in one's family of origin. Psychological and physical health concerns also differentiated between the two groups. CONCLUSIONS: Many variables may differentiate between women incest survivors who self-mutilate from those who do not. A rudimentary checklist to describe the lives of incest survivors who self-mutilate resulted from these findings. The importance of the concept of embodiment is also discussed. PMID- 10695519 TI - Texture response patterns associated with sexual trauma of childhood and adult onset: developmental and recovered memory implications. AB - OBJECTIVE: Reduce texture sensitivity on the Rorschach is proposed as a sequela of early sexual abuse that is unlikely to be contaminated by situational variables. If this conceptualization has merit, texture attributes offer a roadmap for studying vying claims in the recovered memory debate. To explore this possibility, we examined the extent to which intense preoccupation with sexual trauma of childhood and of adult onset was related to reduced texture productivity. METHOD: Texture productivity was measured in 4 groups comprised of 108 patients using the Rorschach. Twenty-seven patients with recovered memory were compared with 27 patients with continuous memory of childhood sexual trauma, 27 post trauma stress patients with sexual trauma of adult onset, and 27 non abused patients. RESULTS: The study replicated previous findings of reduced texture productivity among patients who always remembered sexual trauma of childhood-onset. The same texture deficiency pattern was observed among patients who recovered memory of childhood sexual abuse. This pattern was not observed in Post Traumatic Stress Disorder (PTSD) patients intensely preoccupied with sexual trauma of adult onset despite the fact that they mimicked the recovered memory group in respect to enduring preoccupation with distressing thoughts of sexual abuse. CONCLUSION: The findings indicate that intrusive memories of sexual trauma do not shape patients' response to textural cues on the Rorschach. Variations in texture productivity are primarily moderated by age of trauma onset. Dismissal of claims of recovered memories on the basis of intense sexual preoccupation is not warranted. PMID- 10695520 TI - Risk factors and child sexual abuse among secondary school students in the Northern Province (South Africa). AB - OBJECTIVE: This is an investigation into the risk factors that could discriminate childhood sexual abuse (CSA) from non-abuse in the Northern Province (South Africa). METHOD: 414 students in standard 9 and 10 in three secondary schools in the province filled-in a retrospective self-rating questionnaire in a classroom setting. Questionnaires included modified and adapted questions from the Finkelhor's (1979) Risk Factor Checklist, and asked for physical contact forms of sexual abusive experiences of participants before the age of 17 years with an adult or a person at least 5 years older or a person in a position of power. RESULT: It shows an overall (N = 414) CSA prevalence rate of 54.2%. Only four factors (from eight)-ethnicity not Northern Sotho, mother employed and not as laborer, a stepparent present in the family during childhood, and violence at home not seldom-significantly discriminated CSA from non-abuse. Increase in the number of combination of the four significant factors also increases the probability of the discrimination in a linear manner. CONCLUSION: With some caution, we recommend the four significant risk factors for use while planning preventive strategies against childhood sexual abuse, and a massive campaign against child sexual abuse in the province. More job opportunities should be created in the province. PMID- 10695521 TI - Disclosure of child sexual abuse. PMID- 10695522 TI - Delay in disclosure of childhood rape: results from a national survey. AB - OBJECTIVE: This study sought to gather representative data regarding the length of time women who were raped before age 18 delayed prior to disclosing such rapes, whom they disclosed to, and variables that predicted disclosure within 1 month. METHOD: Data were gathered from 3,220 Wave II respondents from the National Women's Study (Resnick, Kilpatrick, Dansky, Saunders, & Best, 1993), a nationally representative telephone survey of women's experiences with trauma and mental health. Of these, 288 retrospectively reported at least one rape prior to their 18th birthday. Details of rape experiences were analyzed to identify predictors of disclosure within 1 month. RESULTS: Fully 28% of child rape victims reported that they had never told anyone about their child rape prior to the research interview; 47% did not disclose for over 5 years post-rape. Close friends were the most common confidants. Younger age at the time of rape, family relationship with the perpetrator, and experiencing a series of rapes were associated with disclosure latencies longer than 1 month; shorter delays were associated with stranger rapes. Logistic regression revealed that age at rape and knowing the perpetrator were independently predictive of delayed disclosure. CONCLUSIONS: Delayed disclosure of childhood rape was very common, and long delays were typical. Few variables were identified that successfully predicted disclosure behavior, but older age and rape by a stranger were associated with more rapid disclosure. This suggests that the likelihood of disclosure in a given case is difficult to estimate, and predictions based on single variables are unwarranted. PMID- 10695523 TI - Mental health problems among children living in war zones. PMID- 10695524 TI - Post traumatic stress disorder reactions in children of war: a longitudinal study. AB - OBJECTIVE: To establish rates of posttraumatic stress disorder (PTSD) reactions and general mental health problems in children who had experienced war trauma. METHOD: A longitudinal study in the Gaza strip with 234 children aged 7 to 12 years, who had experienced war conflict, at 1 year after the initial assessment, that is, during the peace process. Children completed the Child Post Traumatic Stress Reaction Index (CPTS-RI), while the Rutter A2 and B2 Scales were completed by parents and teachers. RESULTS: The rate of children who reported moderate to severe PTSD reactions at follow-up had decreased from 40.6% (N = 102) to 10.0% (N = 74). 49 children (20.9%) were rated above the cut-off for mental health problems on the Rutter A2 (parent) Scales, and 74 children (31.8%) were above the cut-off on the Rutter B2 (teacher) Scales. The total scores on all three measures had significantly decreased during the 1-year period. The total CPTS-RI score at follow-up was best predicted by the number of traumatic experiences recalled at the first assessment. CONCLUSIONS: PTSD reactions tend to decrease in the absence of further stressors, although a substantial proportion of children still present with a range of emotional and behavioral problems. Cumulative previous experience of war trauma constitutes a risk factor for continuing PTSD symptoms. PMID- 10695525 TI - Model-free functional MRI analysis using Kohonen clustering neural network and fuzzy C-means. AB - Conventional model-based or statistical analysis methods for functional MRI (fMRI) suffer from the limitation of the assumed paradigm and biased results. Temporal clustering methods, such as fuzzy clustering, can eliminate these problems but are difficult to find activation occupying a small area, sensitive to noise and initial values, and computationally demanding. To overcome these adversities, a cascade clustering method combining a Kohonen clustering network and fuzzy, means is developed. Receiver operating characteristic (ROC) analysis is used to compare this method with correlation coefficient analysis and t test on a series of testing phantoms. Results shown that this method can efficiently and stably identify the actual functional response with typical signal change to noise ratio, from a small activation area occupying only 0.2% of head size, with phase delay, and from other noise sources such as head motion. With the ability of finding activities of small sizes stably this method can not only identify the functional responses and the active regions more precisely, but also discriminate responses from different signal sources, such as large venous vessels or different types of activation patterns in human studies involving motor cortex activation. Even when the experimental paradigm is unknown in a blind test such that model-based methods are inapplicable, this method can identify the activation patterns and regions correctly. PMID- 10695526 TI - Quantitative spectral/spatial analysis of phased array coil in magnetic resonance imaging based on method of moment. AB - A new approach for analysis of RF coils in magnetic resonance (MR) experiments is reported. Instead of assuming current distribution in conventional quasi-static algorithm, this approach transforms the coil geometry into an equivalent circuit for complex current calculation. Self and mutual inductance are taken into consideration. Frequency responses of RF coils and transverse magnetic field (B1) maps can be simulated. This approach is especially efficient for phased array coil design for its small matrix size when implemented on computers. Experiments on both single surface coil and phased array coils are consistent with simulation results. Index Terms-Magnetic resonance, method of moment, phased array coil, RF coil. PMID- 10695527 TI - Application of Bayesian inference to fMRI data analysis. AB - The methods of Bayesian statistics are applied to the analysis of fMRI data. Three specific models are examined. The first is the familiar linear model with white Gaussian noise. In this section, the Jeffreys' Rule for noninformative prior distributions is stated and it is shown how the posterior distribution may be used to infer activation in individual pixels. Next, linear time-invariant (LTI) systems are introduced as an example of statistical models with nonlinear parameters. It is shown that the Bayesian approach can lead to quite complex bimodal distributions of the parameters when the specific case of a delta function response with a spatially varying delay is analyzed. Finally, a linear model with auto-regressive noise is discussed as an alternative to that with uncorrelated white Gaussian noise. The analysis isolates those pixels that have significant temporal correlation under the model. It is shown that the number of pixels that have a significantly large auto-regression parameter is dependent on the terms used to account for confounding effects. PMID- 10695528 TI - Performance analysis of maximum intensity projection algorithm for display of MRA images. AB - The maximum intensity projection (MIP) is a popularly used algorithm for display of MRA images, but its performance has not been rigorously analyzed before. In this paper, four measures are proposed for the performance of the MIP algorithm and the quality of images projected from three-dimensional (3-D) data, which are vessel voxel projection probability, vessel detection probability, false vessel probability, and vessel-tissue contrast-to-noise ratio (CNR). As side products, vessel-missing probability, vessel receiver operating characteristics (ROC's), and mean number of false vessels are also studied. Based on the assumptions that the intensities of vessel, tissue, and noise along a projection path are independent Gaussian, these measures are derived and obtained all in closed forms. All the measures are functions of explicit parameters: vessel-to-tissue noise ratio (VTNR) and CNR of 3-D data prior to the MIP, vessel diameter, and projection length. It is shown that the MIP algorithm increases the CNR of large vessels whose CNR prior to the MIP is high and whose diameters are large. The increase in CNR increases with projection path length. On the other hand, all the proposed measures indicate that the small vessels that have low CNR prior to the MIP and small diameters suffer from the MIP. The performance gets worse as projection path length increases. All measures demonstrate a better performance when the vessel diameter is larger. Other properties and possible applications of the derived measures are also discussed. PMID- 10695529 TI - Classifying mammographic mass shapes using the wavelet transform modulus-maxima method. AB - In this article, multiresolution analysis, specifically the discrete wavelet transform modulus-maxima (mod-max) method, is utilized for the extraction of mammographic mass shape features. These shape features are used in a classification system to classify masses as round, nodular, or stellate. The multiresolution shape features are compared with traditional uniresolution shape features for their class discriminating abilities. The study involved 60 digitized mammographic images. The masses were segmented manually by radiologists, prior to introduction to the classification system. The uniresolution and multiresolution shape features were calculated using the radial distance measure of the mass boundaries. The discriminating power of the shape features were analyzed via linear discriminant analysis (LDA). The classification system utilized a simple Euclidean metric to determine class membership. The system was tested using the apparent and leave-one-out test methods. The classification system when using the multiresolution and uniresolution shape features resulted in classification rates of 83% and 80% for the apparent and leave-one-out test methods, respectively. In comparison, when only the uniresolution shape features were used, the classification rates were 72 and 68% for the apparent and leave-one-out test methods, respectively. PMID- 10695530 TI - Classification of malignant and benign masses based on hybrid ART2LDA approach. AB - A new type of classifier combining an unsupervised and a supervised model was designed and applied to classification of malignant and benign masses on mammograms. The unsupervised model was based on an adaptive resonance theory (ART2) network which clustered the masses into a number of separate classes. The classes were divided into two types: one containing only malignant masses and the other containing a mix of malignant and benign masses. The masses from the malignant classes were classified by ART2. The masses from the mixed classes were input to a supervised linear discriminant classifier (LDA). In this way, some malignant masses were separated and classified by ART2 and the less distinguishable benign and malignant masses were classified by LDA. For the evaluation of classifier performance, 348 regions of interest (ROI's) containing biopsy proven masses (169 benign and 179 malignant) were used. Ten different partitions of training and test groups were randomly generated using an average of 73% of ROI's for training and 27% for testing. Classifier design, including feature selection and weight optimization, was performed with the training group. The test group was kept independent of the training group. The performance of the hybrid classifier was compared to that of an LDA classifier alone and a backpropagation neural network (BPN). Receiver operating characteristics (ROC) analysis was used to evaluate the accuracy of the classifiers. The average area under the ROC curve (A(z)) for the hybrid classifier was 0.81 as compared to 0.78 for the LDA and 0.80 for the BPN. The partial areas above a true positive fraction of 0.9 were 0.34, 0.27 and 0.31 for the hybrid, the LDA and the BPN classifier, respectively. These results indicate that the hybrid classifier is a promising approach for improving the accuracy of classification in CAD applications. PMID- 10695531 TI - Confidence maps and confidence intervals for near infrared images in breast cancer. AB - This paper extends basic concepts of statistical hypothesis testing and confidence intervals to images generated by a new procedure for near infrared spectroscopic tomography being developed for use in breast cancer diagnosis. By estimating the covariance matrix of the pixels of an image from data used in the image reconstruction process, confidence maps for statistical tests on individual pixels and confidence intervals for entire images are displayed as an aid to research and clinical personnel interpreting possibly noisy images. The methods are applied to simulated and phantom-based images. PMID- 10695532 TI - Calculation of attenuation factors from combined singles and coincidence emission projections. AB - We have developed a simple method for determining coincidence attenuation correction factors C (the inverse of the total attenuation factors) from collimated singles (SPECT) and coincidence [positron emission tomography (PET)] projections without transmission data. Attenuation-correction factor estimates are determined for individual lines of response (LOR's) independently. The required data can be acquired using a gamma-camera system with coincidence capabilities. A first-order approximation (R) of C for an LOR is given by the product of the singles count rates, taken at each end of the LOR divided by the square of the coincidence count rate. The method was tested using simulated singles and coincidence projections starting with emission and attenuation maps from patient PET scans. Noise and resolution effects were modeled in separate studies. In the noise-free, high-resolution simulations, a scatter plot of the C values versus the corresponding R values for all LOR's produces a well-defined trajectory with little variance. Values of lnR were reconstructed into good quality attenuation maps that compare favorably with the originals. We conclude that the method works well on ideal data. The introduction of noise results in degraded images. In a simulated patient study, lung and outer body boundaries were visible in images produced with 3.2 x 10(4) coincidence counts. PMID- 10695533 TI - Implantation of endovascular stents for hypoplasia of the transverse aortic arch. AB - Hypoplasia of the transverse aortic arch is commonly associated with aortic coarctation. Persistent or recurrent obstruction can occur at this level after successful repair of the native coarcted segment. The purpose of this report is to present a new technique to treat such lesions, namely with implantation of a balloon-expandable stent. This approach was used successfully in 4 children with such hypoplasia occurring after repair of coarctation. Implantation led to both anatomical and physiological relief of obstruction in all. The patients tolerated the procedure, and there were no major adverse events. PMID- 10695534 TI - Cyanotic nephropathy and use of non-ionic contrast agents during cardiac catherization in patients with cyanotic congenital heart disease. AB - BACKGROUND: Chronic cyanosis with its associated rheologic changes is a known risk factor for glomerular nephropathy. Therefore, contrast-induced nephrotoxicity should be an important consideration for angiographers comparable to diabetics. On the other hand, progressions in diagnostic and interventional techniques have led to expanded indications and a more widespread use of x-ray contrast agents. The aim of this study was to investigate the risk of contrast induced nephropathy in the small group of patients with cyanotic heart disease prior to surgical repair. METHODS: We investigated 23 cyanotic patients with an oxygen saturation of 82 (50-92)%, age 25 (5-63) years, and 13 control subjects with atrial septal defect, age 37 (20-66) years. Blood viscosity was measured before and after cardiac catherization. Renal damage was evaluated by selective analysis of urinary proteins and enzymes. RESULTS: Before cardiac catheterization, 48% of the cyanotic patients had a moderate glomerulopathy. Cardiac catherization was performed with 3.0 (1.2-6.8) mls/kg non ionic contrast medium. Only one of the 23 patients (4.3%) with normal urinary analysis before cardiac catheterization showed renal damage, which involved tubular and glomerular function. Elevated blood viscosity in cyanotic patients was slightly reduced by the contrast. None of the acyanotic controls had contrast-induced nephropathy. CONCLUSIONS: The use of non-ionic contrast medium does not worsen cyanotic glomerulopathy. This finding may be due to the reduction of blood viscosity by the application of the contrast medium. The finding of contrast induced nephropathy in one patient underlines the importance of monitoring renal function after cardiac catheterization. PMID- 10695535 TI - Coronary arterial fistulas in childhood. AB - We reviewed 16 patients with coronary arterial fistulas seen between 1976 and 1997, and aged 2 days to 16 years, with a median age of 3.2 years. Only four patients were symptomatic: two had heart failure, one had exertional dyspnoea, and one infective endocarditis. The fistulas originated from the right coronary artery in seven patients, from the left coronary artery in seven, from both coronary arteries in one patient, while the origin was not clearly defined in the final patient. Associated cardiac anomalies were discovered in six patients, with three of the fistulas being diagnosed at the same presentation. Cross-sectional echocardiography had revealed a dilated coronary artery in 7 out of 11 subjects. The ratio of pulmonary to systemic flows ranged between 0.9 to 3.0, with a median of 1.5. Ten patients were referred for corrective surgery without any mortality. Trans-catheter closure was successfully undertaken in one patient, while spontaneous closure of the fistula was noted in two patients. We conclude that coronary arterial fistulas, although rare and potentially serious, are generally treatable. PMID- 10695536 TI - Myocardial perfusion scanning in patients considered for late arterial switch. AB - BACKGROUND: Our aims were to evaluate left ventricular uptake of radionuclide in patients with Mustard's or Senning's procedure, comparing them with patients who had undergone banding of the pulmonary trunk and conversion to the arterial switch. METHODS: Technetium perfusion scans were performed on 27 patients (25 male), aged from 10 to 28 years with a mean of 17.8 years and a standard deviation of 5.8 years, who had undergone Mustard's or Senning's procedure for correction of complete transposition. Of the 27 patients, six had been accepted for staged conversion to an arterial switch. At the time of the study, two of the six patients had undergone completion to the switch and four had undergone banding of the pulmonary trunk with two then proceeding to the arterial switch. Cardiac catheterisation to measure left ventricular pressure was performed in all six patients and scores for left ventricular uptake of isotope were compared with echocardiographic index of the thickness of the left ventricular posterior wall and measurements of left ventricular pressure. RESULTS: Uptake of isotope by the left ventricle was generally poor, but was higher in patients following banding and conversion to the arterial switch, as well as in two patients with native obstruction of the left ventricular outflow tract, and one other who subsequently was found to have pulmonary venous obstruction. There was a positive correlation between the thickness of the left posterior wall in diastole and left ventricular uptake of isotope (r = 0.74, p < 0.05). There also a positive correlation between left ventricular pressure and uptake of the isotope (r = 0.68, p < 0.05). CONCLUSIONS: Uptake of radionuclide by the left ventricle after Mustard's or Senning's procedure for complete transposition appears to reflect ventricular pressure and myocardial mass. A prospective study would be required to determine the predictive ability of such scans regarding the ultimate outcome of conversion to arterial switch, but our initial findings suggest that the technique provides an additional non-invasive method of monitoring left ventricular response to pulmonary arterial banding. PMID- 10695537 TI - Neonatal intrapericardial teratomas: clinical and surgical considerations. AB - Intrapericardial teratomas are rare primary cardiac tumors of infancy and childhood. We describe three neonates with intrapericardial teratomas diagnosed during fetal life and treated after birth. Clinical and anatomic considerations suggest that cardiopulmonary bypass provides for safe tumor dissection and complete excision of the tumor, thereby decreasing the risk of recurrence. PMID- 10695538 TI - The morphologic variability in atrioventricular valvar atresia. PMID- 10695539 TI - A unique case of ventricular isomerism? AB - Isomerism of the atria has often been described with various complex cardiac malformations. As far as is known, a case of 'ventricular isomerism' has never been recorded. Described is a specimen where, on the basis of morphological criterions, there are two right ventricles. PMID- 10695540 TI - Coarctation of the aorta in dizygotic twins. AB - The incidence of congenital heart disease is higher in monozygotic than dizygotic twins, with a higher concordance rate. Although coarctation of the aorta has previously been reported in monozygotic twins, to the best of our knowledge it has not been described in dizygotic twins. We report here such a concurrence in dizygotic twins conceived by in-vitro fertilisation. The finding provides support for both genetic and environmental factors in the aetiology of congenital heart disease. Furthermore, it highlights our lack of data regarding the outcome of in vitro fertilisation. PMID- 10695541 TI - Sudden cardiac death in infancy due to histiocytoid cardiomyopathy. AB - Detailed post-mortem is crucial in infants who die suddenly and without a known cause. We report a rare case of histiocytoid cardiomyopathy with endocardial fibroelastosis, the second case in the world literature. The infant presented with sudden death, but the cardiac histological appearance was initially believed to be caused by Pompes disease. PMID- 10695542 TI - A "friendly" system for pericardial drainage. AB - We describe an alternative to the water-seal system, suitable for drainage of pericardial and eventually pleural effusions, which has been shown to be safe, effective, not painful, and rather "friendly", providing patients with a smoother clinical course. PMID- 10695543 TI - Fetal diagnosis of common arterial trunk with interrupted aortic arch using color power Doppler angiography. AB - We successfully visualized the brachiocephalic arteries and aortic arch in a fetus seen at 19 weeks of gestation with a common arterial trunk and interrupted aortic arch by means of color power Doppler angiography, a new diagnostic development of color Doppler echocardiography. Power Doppler imaging is more sensitive to the state of low flow in fetal vessels, thus providing better visualization of fetal vascular structures from an early gestational stage. PMID- 10695544 TI - Early onset of progressive subaortic stenosis after complete repair of tetralogy of Fallot. AB - Tetralogy of Fallot is often found in association with a wide variety of other cardiac lesions, but is rarely found in association with lesions causing obstruction to the left ventricular inflow or outflow. Subaortic stenosis has only rarely been reported in association with tetralogy of Fallot. We report a patient with Marfan syndrome who underwent repair of tetralogy of Fallot at five years of age. Discrete and progressive subaortic stenosis developed two years after the surgical correction, in a previously normal and unobstructed left ventricular outflow tract. Surgical removal of the acquired fibrous subaortic shelf was successful. Clinical signs of obstruction within the left ventricular outflow tract after surgical repair of tetralogy of Fallot should prompt further investigation to exclude the onset of acquired subaortic stenosis. PMID- 10695545 TI - A most peculiar coronary circulation in a patient with pulmonary atresia and intact ventricular septum. AB - A patient with pulmonary atresia and intact ventricular septum was found to have a right ventricular-dependent coronary circulation. In this infant both coronary arteries lacked their normal proximal connection with the aorta, perhaps the most egregious form of this prejudicial coronary circulation. Even more interesting was a direct collateral vessel originating from the descending thoracic aorta and connecting with the coronary circulation. This patient has undergone bilateral modified Blalock-Taussig shunts, and left ventricular function seems preserved. PMID- 10695546 TI - Complete surgical resection of intrapericardial teratoma in a neonate with compression of the central airways. AB - Intrapericardial teratoma is a rare but recognised cause of respiratory distress in neonates. Patients often present with the compressive effects of the mass within the thorax. Prompt diagnosis should be followed swiftly by surgical resection. We report an unusual case of intrapericardial teratoma in a neonate presenting with collapse of the lung which was successfully treated by surgery. PMID- 10695547 TI - Anterograde double-balloon valvoplasty for treatment of severe valvar aortic stenosis in a preterm baby weighing 1400 grams. AB - We describe our treatment of a premature baby born weighing 1400 g with severe aortic stenosis, with a gradient of 80 mmHg across the valve. Efforts to advance a 6 mm angioplasty catheter into the stenotic aortic valve via the left ventricle failed. Anterograde angioplasty, instead, was performed using two 4 mm coronary angioplasty catheters. Six months subsequent to the intervention, the pressure gradient measured 25 mmHg, and there was no hemodynamically significant aortic insufficiency. PMID- 10695548 TI - Tetralogy of Fallot associated with scimitar syndrome. AB - We present a case of tetralogy of Fallot associated with Scimitar syndrome. The patient was an 11-month old female who underwent successfully total repair of her lesion, including rerouting of the anomalous pulmonary vein to the left atrium. The diagnosis was suspected from the chest x-ray and echocardiography, and confirmed by angiography. To the best of our knowledge only 2 additional cases have previously been reported. PMID- 10695549 TI - Images in congenital heart disease. Transient intimal aneurismal formation during an aortogram using a pigtail catheter. PMID- 10695550 TI - Guidelines for training in paediatric cardiology. PMID- 10695551 TI - Role of Helicobacter pylori infection in extragastroduodenal disorders: introductory remarks. AB - Numerous studies initiated by Warren and Marshall in 1982 confirmed the crucial role of H. pylori infection in the pathogenesis of gastritis, peptic ulcer and possibly also gastric cancer leading to reappraisal of fundamental concept of gastric pathophysiology. These topics were covered, in part, by our previous H. pylori-related symposium I (1995), II (1997) and III (1999) organized in Cracow. H. pylori is one of the most frequent causes of gastroduodenal infection worldwide, resulting in the release of various bacterial and host dependent cytotoxic substances including ammonia, platelet activating factor (PAF), cytotoxins and lipopolysaccharides (LPS) as well as cytokines such as interleukins (IL)-1-12, tumor necrosis factor alpha (TNF(alpha), interferon gamma (INFgamma) and reactive oxygen species (ROS). Recently, several extradigestive pathologies have been linked to H. pylori infection including cardiovascular, cutaneous, autoimmune, esophageal and other diseases such as sideropenic anemia, growth retardation, extragastric MALT-lymphoma etc. The potential role of H. pylori infection in the pathogenesis of these extradigestive disorders has been based on facts that 1) local gastric inflammation may exert systemic effects, 2) chronic infection of gastric mucosa induces immune responses that are able to cause the lesions remote to primary site of infection and 3) H. pylori eradication improves the extradigestive disorders. The aim of present III International Symposium is to provide critical reviews based on personal experience and the available literature about extragastric manifestations of H. pylori infection. The ultimate goal of this symposium is to foster interdisciplinary research and exchange of opinion about the possible involvement of H. pylori in extradigestive pathologies. PMID- 10695552 TI - Helicobacter pylori associated gastric pathology. AB - Helicobacter pylori (HP), undoubtedly, the most common world-wide infection plays an important role in pathogenesis of peptic ulcer. Proof for a causal role for HP in peptic ulcer rests in two major points; 1) the majority of ulcer patients are HP infected and the prevalence of this infection for both gastric ulcer (GU) and duodenal ulcer (DU) is much higher than for gender- and age-adjusted controls and 2) the cure of HP infection dramatically reduces ulcer recurrence. Conclusions regarding the mechanisms by which HP induces peptic ulcer are restricted mainly to studies observing the consequences of its eradication by antibiotics combined with gastric inhibitors or bismuth agents. Several specific virulence factors such as cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) as well as other noxious substances including ammonia, lipopolysaccharide (endotoxin), platelet activating factor (PAF), nitric oxide (NO) and others have been implicated in gastritis and were found to be significantly more frequent in gastric cancer than in gender- and age-matched controls, especially in younger generation. Chronic inflammation, atrophic gastritis, intestinal metaplasia, impaired defense mechanisms combined with hypergastrinemia, deficiency of vitamin C in the stomach , excessive oxygen metabolites and epithelial cell proliferation have been associated with gastric cancer. This multi-step pathway originally proposed by Correa and his colleagues, long before the HP was discovered in the stomach, leads to cancer but may be reversed by eradication of HP. This is, however, a controversial issue because gastric atrophy and intestinal metaplasia may be also caused by other factors such as bile reflux, dietary irritants, and autoimmunity. The implication of HP in MALT-lymphoma is based on the observations that eradication of HP in early stage of low-grade of this tumor leads to complete remission. The significance of HP in non-ulcer dyspepsia remains questionable and requires further studies. PMID- 10695553 TI - Epidemiology of Helicobacter pylori: transmission, translocation and extragastric reservoirs. AB - Although H. pylori infection is endemic and despite more than 10 years of research, the mode and route of transmission remain elusive. This may, in part, be due to the inherent problems of detecting H. pylori noninvasively. The prevalence of infection varies between countries and is closely related to Growth Domestic Product. An age-cohort effect and data from longitudinal studies suggest that the incidence of infection is much higher in children than adults. In developing countries the prevalence of infection is often more than 80% in young adults, in contrast to less than 10% for similar age groups in developed countries. The observations of mosaicism (in the VacA gene) and a panmycytic population structure imply exchange of genetic material either in or outside of the host, which is supported by the increasing recognition of polyclonal infection and suggests that secondary infection occurs after primary acquisition. In addition, in children persistent primary infection may sometimes occur only after previous (repeated) exposure and/or transient colonisation of the gastric mucosa. H. pylori and other gastric Helicobacter spp are always noninvasive, but other human nongastric Helicobacter spp have sometimes been isolated from the systemic circulation in immunocompromised patients. For nonhuman hosts, intestinal Helicobacter spp are thought to translocate more frequently from the colon to the liver. Within the human host, the oral cavity is the principal extragastric reservoir, although case reports suggest that H. pylori may sometimes be found beyond the 2nd part of the duodenum. The hypothesis that H. pylori is a zoonosis or transmitted as coccoid forms by a vector (pets, houseflies) is not supported by recent research showing that H. pylori is entirely unable to support an aerobic or anaerobic metabolism and that coccoid forms are non-viable. H. pylori is primarily acquired in infancy, most probably via the oroorogastric route, from other family members or close contacts encountered after weaning or socialisation. Further studies to support or refute this hypothesis are required. PMID- 10695554 TI - Epidemiological study on Helicobacter pylori infection and extragastroduodenal disorders in Polish population. AB - An association between Helicobacter pylori (H. pylori) infection and extragastroduodenal disorders (EGDD) is still not clear. The aim of the study was to investigate the relationship between H. pylori infection and the symptoms of coronary artery disease (CAD), facial dermatological changes (FDC), gastroesophageal reflux diseases (GERD), and periodontal diseases (PD) in Polish population. The study was performed between 1996-1999 year on 7,060 adult inhabitants of municipal area of Krakow (aged 18-76, mean 46.3 year; 55.8% female, 44.2% male): 2,204 subjects with EGDD and 4,856 without symptoms of EGDD. Each patient responded to a detailed questionnaire under supervision of medical staff. The H. pylori status was assessed non-invasively using urea breath test (UBT) with capsulated low-dose 13C-UBT (38 mg). Exclusion criteria were: recent H. pylori eradication, treatment with PPI, bismuth and/or antibiotics in the last 4 weeks. Four groups of cases with EGDD symptoms were selected. Within each group exclusively only one of studied symptoms was recorded. The study included 328, 138, 688, and 1,050 patients with CAD, FDC, GERD and PD, respectively. For each studied group an age and sex-matched asymptomatic controls were selected (897, 387, 1,083, and 2,489 control patients). RESULTS: Overall H. pylori infection rate was 69,9% (in 71.4% of 2,204 cases and in 69.31% of 4,856 controls). In CAD group: 68% of 328 cases were H. pylori (+ve) vs. 70% H. pylori (+ve) of 897 controls. An association was not significant: OR = 0.93 (95% CI, 0.72-1.20). In 138 of FDC cases, 59% were H. pylori (+ve) vs. 71% H. pylori (+ve) in 387 controls showing the lack of positive association; OR = 0.60 (95% CI, 0.42-0.87). In GERD, 69% of 688 cases were H. pylori (+ve) vs. 73% of 1,083 H. pylori (+ve) controls and negative association was observed; OR=0.80 (95% CI, 0.65-1.00). In 1,050 of PD cases 75% were H. pylori (+ve) vs. 68% H. pylori (+ve) of 2,489 controls; positive association was significant; OR = 1.4 (95% CI, 1.16-1.68). We conclude that in the studied Polish population, no positive association exists between H. pylori positivity and CAD, FDC or GERD possibly due very high overall H. pylori infection rate. The only positive link observed between H. pylori infection and periodontal disease may reflect direct "in situ" H. pylori pathological action of H. pylori in oral cavity. It is not excluded that periodontal diseases may facilitate the H. pylori oro-gastric transmission and colonisation of the bacteria in the digestive tract. PMID- 10695555 TI - The role of Helicobacter pylori in cardiovascular and cerebrovascular diseases. AB - Classical risk factors for cardiovascular and cerebrovascular diseases do not fully coincide with the prevalence of these conditions. Emerging evidences show that new factors may be predisposing for the development of ischemic events. It has been demonstrated that atherosclerosis has a strong inflammatory background; such state of chronic inflammation may be related to the presence of persistent infectious agent. Helicobacter pylori (H. pylori), among other microorganisms, has been extensively investigated for its possible role. Many molecular mechanisms have been hypothesized to explain its eventual action. Epidemiological studies do not exclude a correlation between the infection and cardiovascular and cerebrovascular diseases. Many confounding factors, however, make difficult a definitive evaluation of the huge number of data present in the literature. Moreover, various therapeutic studies have been attempted to show if antibiotic treatment improves prognosis in patients affected by ischemic heart disease. Still, none of these trials focused specifically on the effects of H. pylori eradication on the clinical progression of vascular lesions. PMID- 10695556 TI - Association of Helicobacter pylori infection with coronary heart disease. AB - The role of Helicobacter pylori (HP) as the main etiological factor in gastritis and peptic ulcer disease is undisputable. Gastric mucosal damage caused by HP involves various bacterial and host-dependent toxic substances that have been recently associated with an increased risk of coronary artery disease (CAD), possibly through the activation acute phase response and of procoagulant hemostatic factors. Recent studies showed a close and strong correlation between plasma increments of some cytokines such as IL-6 or TNFalpha and cardiovascular diseases. HP infection induces platelet activation and aggregation that could be the pathogenic explanation of the association between HP infection and CAD. The aim of this study was to determine the seroprevalence of HP infection and antibodies to CagA, an antigen that is expressed by the most virulent HP strains inducing an enhanced gastric inflammatory response, in patients undergoing routine coronary artery examination. We studied 76 patients with CAD and 81 healthy controls patients without significant change in coronary circulation. Angiograms were read by two independent experienced cardiologists blinded to the results of HP status. The presence of serum IgG antibodies to HP and to CagA and plasma interleukin-8 (IL-8) levels was measured by ELISA. In addition plasma C reactive protein fibrinogen, total cholesterol and lipids levels were measured in all studied patients. Seropositivity to HP was found in 81.5 % of cases and in 51% of controls and the difference in prevalence was statistically significant, the odds ratio being 4.3 for Hp patients. Antibody to CagA protein was detected in 47.3% of CAD but only in 28% of healthy controls (OR = 2.3 vs OR = 10). C reactive protein, plasma fibrinogen and total cholesterol were, respectively higher in patients with CAD than in controls. Present data show that there is significant link between CAD and HP infection. The HP infection significantly increases the risk of CAD, especially when both the anti-HP IgG and anti-CagA IgG are considered. Higher prevalence of cytotoxic HP strains might enhance the atherosclerotic process by inducing a persistent, low grade inflammatory response in arterial wall with enhanced synthesis of acute phase reactants. PMID- 10695557 TI - Helicobacter pylori infection and skin diseases. AB - There is increasing evidence for systemic effects of gastric H. pylori infection which may result in extragastrointestinal disorders. This review summarizes the available medical literature up to September 1999, identified through a MEDLINE research including own studies, regarding H. pylori and skin diseases. Due to current knowledge best evidence for a potential link of H. pylori infection exists for chronic urticaria although the data are still conflicting. Thus, the search for H. pylori should be included in the diagnostic management of chronic urticaria. With regard to other skin diseases such as rosacea, hereditary or acquired angioedema due to C1-esterase inhibitor deficiency, systemic sclerosis, Schonlein-Henoch purpura, Sjogren's syndrome, sweet's syndrome, and atopic dermatitis only single of few cases have been reported so far. Thus, we clearly need further randomized, double-blind and placebo-controlled studies including adequate diagnostic schedules, sufficient eradication treatment protocols, confirmation of eradication, and adequate control groups to establish a role of H. pylori in skin diseases. Caution must be taken not to accuse H. pylori as the infectious agent responsible for every disease, particularly since H. pylori infection is very common. Although from an epidemiological and morphological view the skin diseases to which H. pylori has been linked seem to be completely different it is striking that in most of them an autoimmune pathogenesis is suspected or considerable vascular impairment can be found. PMID- 10695558 TI - Helicobacter pylori and its eradication in rosacea. AB - Rosacea is a common condition of unknown etiology usually accompanied by gastrointestinal symptoms and favorably responding to the treatment with antibiotics. This study was designed to examine the prevalence of gastric Helicobacter pylori (Hp) infection verified by 13C-UTB-test, CLO, Hp culture and serology (IgG) in patients with rosacea. Gastroduodenoscopy was combined with pentagastrin secretory test and antral and fundic biopsy samples were taken for histological evaluation (the Sydney system). Blood samples were also taken for the determination of plasma gastrin using RIA and plasma interleukin (IL)-8 and tumor necrosis factor alpha (TNFalpha) using ELISA. This study was performed in 60 patients, 31-72 year old, with visible papules and pustules associated with erythema and flushing on the face and on 60 age- and gender-matched patients without any skin diseases but with similar as in rosacea gastrointestinal symptoms but without endoscopic changes in gastroduodenal mucosa (non-ulcer dyspepsia - NUD). The Hp prevalence in rosacea patients was about 88 % as compared to 65% in control NUD patients. Among rosacea patients, 67% were cytotoxin associated gene A (CagA) positive, while in NUD patients only 32% were CagA positive. Rosacea patients showed gastritis with activity of about 2.1 in antrum and 0.9 in the corpus of the stomach while those with NUD only mild gastritis with activity of approximately 1.0) confined to the antrum only. Following initial examination, typical 1 wk anti-Hp therapy including omeprazole (20 mg bd.), clarithromycin (500 mg bd.) and metronidazol (500 mg bd.) was carried out. After eradication, 51 out of 53 treated rosacea patients became Hp negative. Within 2-4 weeks, the symptoms of rosacea disappeared in 51 patients, markedly declined in 1 and remained unchanged in 1 other subject. A dramatic reduction in activity of gastritis (to 0.3 in antrum and to 0.1 in corpus) was observed. Basal plasma gastrin decreased from 48 +/- 5 pM before to 17+/-3 pM after eradication, while pentagastrin-induced maximal (MAO) declined, respectively, from about 16.6 +/- 4.2 to 8.5 +/- 1.8 mmol/h. Plasma TNFalpha and IL-8 were reduced after the therapy by 72% and 65%, respectively. We conclude that: 1) Rosacea is a disorder with various gastrointestinal symptoms closely related to gastritis, especially involving the antrum mucosa, with Hp expressing cagA in the majority of cases and elevated plasma levels of TNFalpha and IL-8; 2) The eradication of Hp leads to a dramatic improvement of symptoms of rosacea and reduction in related gastrointestinal symptoms, gastritis, hypergastrinemia and gastric acid secretion; and 3) Rosacea could be considered as one of the major extragastric symptoms of Hp infection probably mediated by Hp-related cytotoxins and cytokines. PMID- 10695559 TI - Helicobacter pylori lipopolysaccharide-mediated gastric and extragastric pathology. AB - Lipopolysaccharides (LPS) are a family of toxic phosphorylated glycolipids in the outer membrane of Gram-negative bacteria, including Helicobacter pylori, and are composed of a lipid moiety (termed lipid A), a core oligosaccharide, and a polymeric O-specific polysaccharide chain. Compared with LPS of other bacteria, H. pylori LPS and lipid A induce low immunological activities in a range of test systems. Nevertheless, these reduced levels of LPS-induced cytokines and toxic oxygen radicals can contribute, with those induced by bacterial proteins, to the H. pylori-associated inflammatory response. Whether the ability of H. pylori LPS to induce low production of both procoagulant activity and plasminogen activator inhibitor type 2 by human mononuclear cells contributes to localized inflammatory responses alone and, in addition, play a role in extragastric pathology remains an open question. The core oligosaccharide of H. pylori LPS, in part with a 25 kDa protein adhesin, mediates the binding of the bacterium to the host glycoprotein laminin, and hence interferes with gastric cell receptor-laminin interaction in the basement membrane. Also affecting mucosal integrity, the core sugars of certain H. pylori strains, particularly those associated with gastric ulceration, have been implicated in pepsinogen induction, but this is a strain dependent phenomenon. Of particular interest, the O-chains of a large proportion of H. pylori strains mimic Lewis (Le) antigens. Although investigations have focussed on the role of these antigens in H. pylori-associated autoimmunity, which remains to be unequivocally established, other pathogenic consequences of Lewis mimicry are becoming apparent. Expression of Lewis antigens may be crucial for H. pylori colonization and adherence and, by aiding bacterial interaction with the gastric mucosa, thereby aid delivery of secreted products, and hence influence the inflammatory response. PMID- 10695560 TI - Immunology of Helicobacter pylori infection. AB - We have tried to answer several questions in this article, dealing with: ontogenesis of the immune response, presentation of H. pylori antigens to immune cells, systemic vs local immune response, cytokine Th1/Th2 configuration, the role of cytokines, especially represented during H. pylori infection, mimicry phenomena, extragastroduodenal sites and manifestations and finally, vaccine development. The new achievements in the vaccine field, also of Polish groups, were underlined. PMID- 10695561 TI - The infection by Helicobacter pylori strains expressing CagA is highly prevalent in women with autoimmune thyroid disorders. AB - H. pylori infection is putatively associated with extra-digestive disorders and may also play a role in the development of autoimmune thyroid diseases (ATD). It was recently found that monoclonal antibodies to an H. pylori strain with cagA positivity reacted with follicular cells of the thyroid gland, and that an H. pylori organism possessing the cag pathogenicity island carried a gene encoding for an endogenous peroxidase. The aims of this study was (1); To ascertain whether the infection by strains endowed with an increased inflammatory potential (those expressing CagA) could further enhance the risk of developing ATD (2); To verify the possible existence of an immune cross-reactivity between autoantibodies to peroxidase and thyroglobulin and H. pylori antigens (3). To establish whether thyroid colloid antigens could cross-react with an anti-H. pylori serum. The study was partly designed retrospectively. We examined 41 consecutive women with ATD, and, as a control, 33 consecutive age- and socio economic class-matched women without autoimmune thyroid disorders, living in the same area as patients, occurred at the same institution in the same period (six months). Both patients and controls were examined serologically for H. pylori infection and CagA status by Western blotting. Some serum samples were absorbed with H. pylori to determine whether the antibody levels decreased. Colloid proteins were resolved electrophoretically and matched with a hyperimmune serum raised in rabbits against a CagA-positive H. pylori. Thirty-two patients (78.0%) tested seropositive for H. pylori infection, vs. 16 controls (48.4%) (P = 0.008, OR = 3.78, RR = 1.61). The prevalence of anti-CagA antibodies was 71.8% in infected patients, and 50% in infected controls (P = 0.161, n.s.). The overall prevalence of infection by CagA-positive H. pylori was significantly higher in patients with ATD (23/41, or 56.0%) than that in controls (8/33, or 24.2%) (P = 0.006, OR = 3.99, RR = 2.31). The other tests gave negative or inexplicable results. IN CONCLUSION: CagA-positive H. pylori infection increases the risk of ATD development. PMID- 10695562 TI - CagA-positive Helicobacter pylori infection may increase the risk of food allergy development. AB - The aim of this study was to test whether patients with symptomatic food allergy and significant levels of immunoglobulin E (IgE) to alimentary antigens were more likely infected by H. pylori, especially by strains expressing the CagA protein, with respect to controls. A group of 38 patients with symptomatic food allergy and 53 age-matched controls were examined serologically for H. pylori infectious status, and for CagA seropositivity. IgE to alimentary allergens were measured by a commercial kit. The prevalence of H. pylori infection in patients with food allergy and controls was similar (42.1%, and 48.3%, respectively). However, anti CagA antibodies in H. pylori-infected persons were detected in 62.5% of patients with food allergy, and 28% of controls (P = 0.030, odds ratio = 4.29). The mean level of IgE to the most common alimentary antigens in serum samples from infected patients with anti-CagA antibodies was significantly higher than in CagA negative infected patients: 3.28 kU/L (SD 3.93), vs. 1.99 kU/L (SD 1.53), P = 0.002, 95% confidence interval = 0.61 to 2.53). Infection by CagA-positive H. pylori increases the risk of developing food allergy. PMID- 10695563 TI - The role of CagA status in gastric and extragastric complications of Helicobacter pylori. AB - Two major markers of virulence have been described in H. pylori. The first is a secreted protein (VacA) that is toxic to human cells in tissue culture. This cytotoxin causes vacuolation of epithelial cells in vitro and induces epithelial cell damage in mice. The second is a 40-Kb pathogenicity island for which the gene cagA (cytotoxin-associated gene A) is a marker. Approximately 60% of H. pylori isolates in Western countries are cagA+. The protein encoded by cagA+ has a molecular weight of 120-140 kDa and exhibits sequence heterogeneity among strains isolated from Western and Eastern countries. Although no specific function has been identified for CagA, there is increasing evidence that cagA+ strains are associated with increased intensity of gastric inflammation and increased mucosal concentration of particular cytokines including interleukin 8. Inactivation of picB (Hp 0544) or any of several other genes in the cag island ablates the enhanced IL-8 secretion of human gastric epithelial cells in tissue culture. Furthermore, persons colonized with cagA+ strains have an increased risk of developing more severe gastric diseases such as peptic ulcer and distal (non cardia) gastric cancer than those harboring cagA- strains. We investigated the role of cagA status in both gastroduodenal and extragastroduodenal disease with H. pylori. Among the diseases limited to the antrum and body of the stomach and the duodenum, we demonstrated a correlation between CagA seropositivity and peptic ulcer disease. We also showed correlation between distal gastric cancer rated and CagA prevalence in populations in both developed and developing countries. In addition, we found that for several Asian populations, the relationship between CagA seropositivity and gastroduodenal diseases was complex. For extragastroduodenal diseases, our results confirmed previous reports that demonstrated that CagA status did not play a role in diseases such as rheumatoid arthritis and hyperemesis gravidarum. However, we found a clear negative association between the presence of a positive response to CagA and esophageal diseases. Therefore, CagA seropositivity (and thus gastric carriage) is associated with increased risks of certain diseases (involving the lower stomach and duodenum) and decreased risks of GERD and its sequelae. This apparent paradox can best be explained by differences in the interaction of cagA+ and cagA- strains with their hosts. PMID- 10695564 TI - The carcinogenic effect of H. pylori on the gastric epithelial cell. AB - The recent demonstration in animal models that H. pylori alone may be capable of inducing intestinal-type gastric cancer, and that H. felis can accelerate gastrin induced gastric neoplasia has stimulated research on examining the mechanisms of H. pylori-associated carcinogenesis in humans. Several mechanisms are currently under investigation, including the dysregulation of the gastric epithelial cell cycle, the formation of DNA adducts, the generation of free radicals, alterations in growth factor secretion and cytokines, and the effects of decreased gastric acid secretion. This review will examine the relevant evidence acquired from human tissue studies, animal models and cell culture systems in an attempt to explore these pathways, and to evaluate the mechanisms by which H. pylori may cause gastric cancer. PMID- 10695565 TI - Role of gastrin in gastric cancerogenesis in Helicobacter pylori infected humans. AB - Numerous epidemiological studies demonstrated the association between Helicobacter pylori (H. pylori) infection and gastric cancer but the mechanism of the involvement of H. pylori in gastric cancerogenesis remains virtually unknown. This study was designed to determine the seropositivity of H. pylori and cytotoxin associated gene A (CagA), serum gastrin and gastric lumen gastrin levels under basal conditions and following stimulation with histamine in gastric cancer patients and controls. 100 gastric cancer patients aging from 21 to 60 years and 300 gender- and age-adjusted controls hospitalized with non-ulcer dyspepsia (NUD) entered this study. 13C-Urea Breath Test (UBT), serum immunoglobulin (IgG) antibodies to H. pylori and CagA were used to assess the H. pylori infection and serum levels of IL-1beta, IL-8 and TNFalpha were measured by enzyme-linked immunosorbent assay (ELISA) to evaluate the degree of gastric inflammation by H. pylori . Gastrin-17 mRNA and gastrin receptors (CCK(B)) mRNA expression in gastric mucosal samples taken by biopsy from the macroscopically intact fundic and antral mucosa as well as from the gastric tumor was determined using RT-PCR. The overall H. pylori seropositivity in gastric cancer patients at age 21-60 years was about 92%, compared, respectively, to 68%, in controls. A summary odds ratio (OR) for gastric cancer in H. pylori infected patients was about 5.0 . The H. pylori CagA seropositivity in gastric cancer patients was about 58.5% compared to 32.4% in controls, giving the summary OR for gastric cancer in CagA positive patients about 8.0. The prevalence of H. pylori- and H. pylori CagA-seropositivity was significantly higher in cancers than in controls, irrespective of the histology of gastric tumor (intestinal, diffuse or mixed type). Median IL-1beta and IL-8 reached significantly higher values in gastric cancer patients (9.31 and 30.8 pg/ml) than in controls (0.21 and 3.12, respectively). In contrast, median serum gastrin in cancers (as total group) was several folds higher (62.6 pM) than in controls (19.3 pM). Also median luminal gastrin concentration in gastric cancer patients was many folds higher (310 pM) than in controls (20 pM). This study shows for the first time that cancer patients are capable of releasing large amounts of gastrin into the gastric lumen to increase luminal hormone concentration to the level that was recently reported to stimulate the growth of H. pylori. There was no any correlation between plasma gastrin levels and gastric luminal concentration of gastrin suggesting that: 1) luminal gastrin originates from different source than plasma hormone, most probably from the cancer cells, 2) cancer cells are capable of expressing gastrin and releasing it mainly into the gastric juice and 3) the gastric cancer cells are equipped with gastrin-specific (CCK(B)) receptor so they exhibit the self growth promoting activity in autocrine fashion. This notion is supported by direct detection of gastrin mRNA and gastrin receptor (CCK(B)-receptors) mRNA using RT-PCR in cancer tissue. To our knowledge this is the first study showing an important role of gastrin as self-stimulant of cancer cells in patients infected with H. pylori. Basal and histamine maximally stimulated acid outputs were significantly lower in gastric cancer patients than in controls despite of enhanced gastrin release, particularly in cancer patients and this might reflect the mucosal inflammatory changes (increased serum levels of proinflammtory interleukins - IL-1beta and IL-8), that are known to increase gastrin release. We conclude that: 1) H. pylori infected patients, particularly those showing CagA seropositivity, are at greatly increased risk of development of gastric cancer, 2) H. pylori-infected cancer patients produce significantly more IL-1beta and IL 8 that might reflect an H. (ABSTRACT TRUNCATED) PMID- 10695566 TI - Detection of Helicobacter in the liver of patients with chronic cholestatic liver diseases. AB - Helicobacter species were identified in human liver tissues by PCR. Biopsies were obtained from patients with primary sclerosing cholangitis, primary biliary cirrhosis and noncholestatic liver cirrhosis. One set of Helicobacter genus specific primers and two different primer sets for Helicobacter pylori were used in the PCR-assays. Using Helicobacter genus-specific primers 80% (8/10) of patients with primary sclerosing cholangitis and 90% (9/10) of patients with primary biliary cirrhosis were positive. Seven of these 17 samples were positive using two different primers for H. pylori and Southern blot hybridization. Among the non-cholestatic liver cirrhosis controls, only one sample was positive in the Helicobacter genus-specific PCR-assay. Significantly higher values of alkaline phosphatases and prothrombin complex was found for the patients positive for Helicobacter genus. In conclusion, gene sequences of Helicobacter species and H. pylori were detected in human liver tissue using PCR and DNA hybridization in patients with a cholestatic liver disease, but rarely in noncholestatic liver cirrhosis. PMID- 10695567 TI - Preventing traumatic brain injury: an innovative approach to outcomes assessment. AB - The purpose of this study was to investigate the usefulness of the state traffic violations database as an outcome measure for the Young Traffic Offenders Programme, a 1-day educational injury prevention programme for young people with speeding offences. The database provided a fine-grained, cumulative record of each driver's traffic offences and sanctions applied. A comparison of 92 programme participants and 87 non-treated individuals showed no significant between-group difference in rate of convictions after the target programme date, but each group declined significantly in rate of convictions over time. Younger males and individuals with a higher number of convictions initially were more likely to have higher conviction rates after the target date. These findings underscore the need to improve prevention programmes and highlight the potential usefulness of existing public datasets for outcome evaluation. PMID- 10695568 TI - Cognitive orientation in rehabilitation and neuropsychological outcome after traumatic brain injury. AB - This study evaluated the ability of the Orientation Log (O-Log) to predict cognitive outcome at rehabilitation discharge, as well as future neuropsychological outcome. The hypothesis was that patients who demonstrated better orientation upon admission would achieve superior functional cognitive outcome at discharge and on subsequent neuropsychological assessment. Sixty individuals receiving inpatient rehabilitation following a new-onset TBI participated. Orientation data was collected using the O-Log during morning bedside rounds. Outcome data was collected at 6 and 12 months post-injury. Significant correlations were found between the O-log and measures of memory, executive functioning, basic verbal skills, and estimated intellectual ability. When compared to the other predictor variables, step-wise multiple regression analyses revealed that the minimum O-Log score was the primary significant predictor of performance on six neuropsychological and functional outcome measures. Results of this study suggest that evaluating orientation with the O Log during acute rehabilitation may reflect level of injury severity and aid in predicting cognitive outcome. PMID- 10695569 TI - Cultural variations in the understanding of traumatic brain injury and brain injury rehabilitation. AB - Little is known about how people from different cultures experience and understand traumatic brain injury (TBI) and the process of brain injury rehabilitation. The Brain Injury Rehabilitation Unit (BIRU) undertook a qualitative project to research cultural variations in the understanding of TBI and the rehabilitation process, interviewing 39 people with TBI and family members from Italian, Lebanese and Vietnamese backgrounds. The focus was on the reporting of sequelae of the TBI; valued qualities of service providers; barriers to effective communication; the role of the families in the rehabilitation process; and the experience of social stigma. Findings suggest there is a universal experience of TBI that transcends individual cultures. Study participants valued attentiveness, friendliness and guidance from rehabilitation staff. Family support was not always available to the person with TBI due to family conflict. Generally, people with TBI and family members valued the assistance of health interpreters facilitating their communication with rehabilitation staff. People with TBI from all three cultures experienced problems of stigma and social isolation. The findings have a number of implications for how brain injury rehabilitation staff can approach service provision to people from diverse cultural backgrounds. PMID- 10695570 TI - Sitting balance following brain injury: does it predict outcome? AB - Balance dysfunction is commonly observed following traumatic brain injury. There are many proposed predictors of functional outcome in the traumatic brain injury population. It was hypothesized that the degree of balance dysfunction on admission to rehabilitation would be a significant predictor of the need for assistance at discharge, as measured by the Functional Independence Measure (FIM). This study involved 237 cases of traumatic brain injury patients admitted to a rehabilitation unit between November 1989 and September 1996. Using a multiple regression model, controlling for age, initial Glasgow Coma Score (GCS), rehabilitation admission strength, sitting balance and standing balance, it was found that the degree of impairment in sitting balance at admission to rehabilitation was a significant predictor of Discharge FIM-Total (FIM-T) score (p < 0.0001) and also of selected elements from the Discharge FIM-Motor (FIM-M) score (p < 0.0005). The combination of age, initial admission GCS, rehabilitation admission strength, standing balance and sitting balance accounted for 29% of the variance in the Discharge Total FIM score. Among these, sitting balance was the second most powerful predictor of both selected elements of the Discharge FIM motor score and discharge FIM-T. Sitting balance predictive capacity was exceeded in power only by age. Impairments in sitting balance appear to have a significant impact on functional outcome. Emphasis on unique rehabilitation techniques to treat balance dysfunction in the adult TBI population is warranted. PMID- 10695571 TI - Kinematic analysis of tongue movements in dysarthria following traumatic brain injury using electromagnetic articulography. AB - Electromagnetic articulography (EMA), a technique that uses alternating magnetic fields to track the movement of miniature receiver coils affixed to the articulators, was used to assess the speed and accuracy of tongue movements exhibited by an individual with dysarthria following severe traumatic brain injury (TBI). Three receiver coils were attached to the TBI subject's tongue and the movements of these coils were recorded during five productions of three single syllable real words consisting of the lingual consonants /t, s, k/ in the word-initial position. A non-neurologically impaired adult male served as a control subject. A range of kinematic parameters was analysed from the consonant productions including the movement trajectories, velocity, acceleration, distance, and duration of tongue movements. Examination of the complex interactions between the kinematic parameters recorded for the TBI subject revealed a disturbance in the 'control' of tongue speed rather than a disturbance in speed per se, as it was found that the TBI subject exhibited difficulty in decelerating his tongue movements appropriately on the approach up to the palate during consonant production. The difficulty noted in deceleration resulted in inaccurate tongue movements that overshot the point of intent (in the case of /t/) and may have been instrumental in reducing the length of time that the tongue remained at the palate (in the case of /s/ and /k/) in comparison to the control subject. The disturbances identified in the kinematic parameters recorded provided objective insights into the nature of the articulatory disturbances responsible for the deviant speech feature, consonant imprecision, perceived in the TBI subject's speech. The study stresses the importance of examining a range of kinematic parameters and the interactions between these parameters in attempting to determine the nature of articulatory disturbances exhibited by individuals with dysarthria following TBI. PMID- 10695572 TI - Coping style and post-traumatic stress disorder following severe traumatic brain injury. AB - There is increasing evidence that a proportion of severe traumatically brain injured (TBI) patients do suffer post-traumatic stress disorder (PTSD). The aim of this study was to investigate the predictors of PTSD following severe TBI in a sample of 96 patients who sustained a severe TBI, of whom 27% satisfied diagnostic criteria for PTSD. The Post-traumatic Stress Disorder Interview, the Coping Style Questionnaire, and the Functional Assessment Measure was administered to these patients 6 months after hospital discharge. Avoidant coping style, behavioural coping style, and a history of prior unemployment were the significant predictors of PTSD severity. These findings indicate that reduction of PTSD and management of severe TBI may be facilitated by teaching patients more adaptive coping strategies. PMID- 10695573 TI - Emergency department visits associated with traumatic brain injury: United States, 1995-1996. AB - The purposes of this study were to provide a national estimate of the incidence of traumatic brain injuries (TBIs) seen in emergency departments (EDs), but not requiring hospitalization and to determine the causes of these injuries. Using the Centers for Disease Control and Prevention case definition of TBI, ED data was analysed from the National Hospital Ambulatory Medical Care Survey (1995 1996). The average overall incidence rate of TBI-related ED visits for persons who were not hospitalized was 392/100,000 population per year, or 1,027,000 visits to hospital EDs in the US each year. This estimate is nearly twice (392 vs. 216) the previously estimated incidence rate, which was based on data from the 1991 National Health Interview Survey Injury Supplement. It was found that the highest incidence rate occurred among children aged 0-14 years, the rate for males was higher than for females, and the primary reported causes of these injuries were 'falls', motor vehicle-related causes, and 'struck by an object'. Although often considered 'mild' TBIs, these injuries can lead to significant cognitive and emotional impairment. Thus, continued surveillance of TBI-related ED visits is an important part of a comprehensive TBI prevention strategy. PMID- 10695574 TI - Electronic memory aids for outpatient brain injury: follow-up findings. AB - The introduction of highly portable computers extends the range of tools potentially useful to persons with functional impairments of prospective memory resulting from brain injury. This study reviews the experience of 12 patients with brain injury undergoing outpatient treatment using palmtop computers to assist with memory dependent activities in their everyday lives. During the initial supervised trial period, each was provided a palmtop computer based memory aid capable of generating audible and visible reminder cues. Subsequently, patients were contacted for follow-up between 2 months and 4 years after initial trial usage, and surveyed as to the utility of the computer. Nine patients found palmtop computers were useful during supervised trials. Seven of nine patients actually continued to use such devices after the usage trials had ended. Experience with this technology has shown it to be useful in a high proportion of patients for assisting with memory dependent functions. PMID- 10695575 TI - Neuropsychological assessment of a potential "euthanasia" case: a 5 year follow up. AB - McMillan reported a neuropsychological assessment procedure which was used to determine whether or not there was evidence for sentience in a young woman who had been rendered tetraplegic and anarthric as a result of a road traffic accident. An application to court had been made to withdraw feeding and this was supported by medical evidence which gave the view that the individual was functioning little beyond vegetative state, had a poor quality of life and had little prospect of further recovery. Evidence for an ability to communicate reliably was found including for a wish to continue living, and as a consequence the application to court was withdrawn. This paper describes further recovery 2-4 years after the original assessment (i.e. 4-6 years post-injury). At follow-up, she remained dependent for all care, but was now feeding orally and was talking. She could learn new information, some of which she retained for at least 12 months and had greater insight into her condition. She now reported low mood and some pain. As before, she consistently reported a wish to live. The implications of the follow-up are discussed in the context of assumptions made about quality of life and decision making about euthanasia in brain injured people who are severely disabled, but are not in a vegetative state. PMID- 10695576 TI - Hypercholesterolemia in children. PMID- 10695577 TI - Family physicians should be experts in palliative care. PMID- 10695578 TI - Neurologic complications of Epstein-Barr virus infection. PMID- 10695579 TI - Patient compliance: in search of the real question in diabetes care. PMID- 10695580 TI - Comment on the family physician as hospitalist. PMID- 10695581 TI - Dietary therapy for children with hypercholesterolemia. AB - Accumulating evidence clearly shows that atherosclerosis begins in youth. The National Cholesterol Education Program (NCEP) has recommended that children at high risk of developing coronary artery disease as adults be screened so that those with elevated low-density lipoprotein (LDL) cholesterol levels can be treated, primarily by modification of diet. The initial approach to these youthful patients is to use the NCEP step I diet. This diet provides calories and nutrients that support normal growth and development, but limits saturated fat and total fat intake to no more than 10 and 30 percent of total calories, respectively, and cholesterol intake to no more than 100 mg per 1,000 kcal per day, to a maximum of 300 mg. If the goal of reducing the LDL cholesterol level to below 130 mg per dL (3.35 mmol per L) is not achieved, the more restrictive step II diet should be initiated. However, the step II diet may not provide sufficient calories and nutrients to support normal growth and development; therefore, trained nutritionists may be required to effectively manage a child on this diet. PMID- 10695582 TI - Evaluation of overuse elbow injuries. AB - The evaluation of elbow pain can be challenging because of the complexity of the joint and its central location in the upper extremity. Diagnosing the injury correctly requires an understanding of the anatomy of the elbow, which includes three articulations, two ligament complexes, four muscle groups and three major nerves. The history should be directed at pinpointing the location of symptoms and the activities that cause the patient's pain. It is important to identify the specific musculotendinous structures that are at risk for overuse or have been injured through overuse. Mechanical symptoms are indicative of intra-articular pathology, whereas neurologic symptoms are characteristic of nerve entrapment syndromes. Physical examination of the elbow and related structures should confirm the diagnosis. Only a minority of patients require diagnostic studies. Basic treatment principles are described by the acronym PRICEMM: protection, rest, ice, compression, elevation, medication and modalities (physical therapy). Surgical consultation is warranted in selected patients. PMID- 10695583 TI - Recognizing an index case of tuberous sclerosis. AB - Tuberous sclerosis is the most common neurocutaneous syndrome after neurofibromatosis. Dermatologic manifestations may be the only clues the family physician has to the diagnosis of the disorder, which is also marked by childhood seizures and mental retardation. Characteristic signs of tuberous sclerosis vary widely in severity and can include hypopigmented "ash-leaf spots," fibrous plaques on the forehead, angiofibromas on the face (adenoma sebaceum), a shagreen patch on the lower back and fibromas of the nails. Computed tomographic scanning or magnetic resonance imaging reveal subependymal nodules or cortical "tubers" in the brain. Associated cardiac, retinal, renal and pulmonary pathology can increase morbidity and mortality. Genetic counseling is helpful but has limited use because of the variation in genetic expression and the frequency of new gene mutations that cause this disorder. PMID- 10695584 TI - Urinary tract infections during pregnancy. AB - Urinary tract infections are common during pregnancy, and the most common causative organism is Escherichia coli. Asymptomatic bacteriuria can lead to the development of cystitis or pyelonephritis. All pregnant women should be screened for bacteriuria and subsequently treated with antibiotics such as nitrofurantoin, sulfisoxazole or cephalexin. Ampicillin should no longer be used in the treatment of asymptomatic bacteriuria because of high rates of resistance. Pyelonephritis can be a life-threatening illness, with increased risk of perinatal and neonatal morbidity. Recurrent infections are common during pregnancy and require prophylactic treatment. Pregnant women with urinary group B streptococcal infection should be treated and should receive intrapartum prophylactic therapy. PMID- 10695585 TI - Treatment of psoriasis: an algorithm-based approach for primary care physicians. AB - Psoriasis is characterized by red, thickened plaques with a silvery scale. The lesions vary in size and degree of inflammation. Psoriasis is categorized as localized or generalized, based on the severity of the disease and its overall impact on the patient's quality of life and well-being. Patient education about the disease and the treatment options is important. Medical treatment for localized psoriasis begins with a combination of topical corticosteroids and coal tar or calcipotriene. For lesions that are difficult to control with initial therapy, anthralin or tazarotene may be tried. The primary goal of therapy is to maintain control of the lesions. Cure is seldom achieved. If control becomes difficult or if psoriasis is generalized, the patient may benefit from phototherapy, systemic therapy and referral to a physician who specializes in the treatment of psoriasis. PMID- 10695586 TI - Pathologic gambling. AB - Pathologic gambling and problem gambling affect approximately 5 to 15 million Americans and are common in young people. The community-minded family physician is in a good position to identify and assist patients who have gambling-related problems and thereby prevent or treat the resultant personal, family and social disruptions. Provider and community education about the depth and breadth of this condition is crucial for the identification and treatment of a growing problem. As with many psychologic conditions, identification of the disorder and treatment of the patient by the family physician comprise the primary treatment. Screening tools, treatment programs and self-help groups provide additional resources for the family physician. An illustrative case report demonstrates the importance of heightened awareness of and screening for this common condition. PMID- 10695587 TI - Managing pain in the dying patient. AB - End-of-life care can be a challenge requiring the full range of a family physician's skills. Significant pain is common but is often undertreated despite available medications and technology. Starting with an appropriate assessment and following recommended guidelines on the use of analgesics, family physicians can achieve successful pain relief in nearly 90 percent of dying patients. Physicians must overcome their own fears about using narcotics and allay similar fears in patients, families and communities. Drugs such as corticosteroids, antidepressants and anticonvulsants can also help to alleviate pain. Anticonvulsants can be especially useful in relieving neuropathic pain. Side effects of pain medications should be anticipated and treated promptly, but good pain control should be maintained. The physical, psychologic, social and spiritual needs of dying patients are best managed with a team approach. Home visits can provide comfort and facilitate the doctor-patient relationship at the end of life. PMID- 10695588 TI - 1999 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with HIV: Part III. Prevention of disease recurrence. United States Public Health Service/Infectious Diseases Society of America. PMID- 10695589 TI - Photo quiz. Palpable shin lesions. PMID- 10695590 TI - Setting limits on demanding patients. PMID- 10695591 TI - ACC/AHA guidelines for ambulatory ECG. American College of Cardiology/American Heart Association. PMID- 10695592 TI - Risk factors for a causal intermediate and an endpoint: reconciling differences. AB - When a risk factor influences the intermediary but not progression to the endpoint, it has been shown that the relative risk estimate for the causal intermediate is identical to that for the endpoint under a single pathway framework. When there are multiple pathways, the relative risk estimate for the endpoint is reduced. The authors examine how the reduction of the effect size from a risk factor's association with the causal intermediate to that with the endpoint relates to the proportion of endpoint cases arising through other pathways, and the measure of effect used. For multiple pathways, all measures of effect are reduced and the reduction increases as the proportion of endpoint cases arising through other pathways increases. For single pathways, the relative rate ratio and odds ratios are reduced. In particular, the reduction in the odds ratio may be dramatic because of the commonness of causal intermediates relative to the endpoint. Comparisons of causal intermediate studies with those for the endpoint should consider the influences of multiple pathways, the prevalence of the causal intermediate, the measure of effect used, and the multiple effects a risk factor may have along the pathway when interpreting the differences observed across the causal chain. PMID- 10695593 TI - A multiethnic cohort in Hawaii and Los Angeles: baseline characteristics. AB - The authors describe the design and implementation of a large multiethnic cohort established to study diet and cancer in the United States. They detail the source of the subjects, sample size, questionnaire development, pilot work, and approaches to future analyses. The cohort consists of 215,251 adult men and women (age 45-75 years at baseline) living in Hawaii and in California (primarily Los Angeles County) with the following ethnic distribution: African-American (16.3%), Latino (22.0%), Japanese-American (26.4%), Native Hawaiian (6.5%), White (22.9%), and other ancestry (5.8%). From 1993 to 1996, participants entered the cohort by completing a 26-page, self-administered mail questionnaire that elicited a quantitative food frequency history, along with demographic and other information. Response rates ranged from 20% in Latinos to 49% in Japanese Americans. As expected, both within and among ethnic groups, the questionnaire data show substantial variations in dietary intakes (nutrients as well as foods) and in the distributions of non-dietary risk factors (including smoking, alcohol consumption, obesity, and physical activity). When compared with corresponding ethnic-specific cancer incidence rates, the findings provide tentative support for several current dietary hypotheses. As sufficient numbers of cancer cases are identified through surveillance of the cohort, dietary and other hypotheses will be tested in prospective analyses. PMID- 10695595 TI - Invited commentary: the challenge of multi-center cohort studies in the search for diet and cancer links. PMID- 10695594 TI - Calibration of the dietary questionnaire for a multiethnic cohort in Hawaii and Los Angeles. AB - The performance of the dietary questionnaire used in a multiethnic cohort study in Hawaii and Los Angeles was assessed in a calibration substudy that compared diet reported from the questionnaire with three 24-hour dietary recalls. For the calibration substudy, subjects from each of eight subgroups defined by sex and ethnic group (African-American, Japanese-American, Latino, and White) were chosen randomly from among the cohort members, and each participant's previous day's diet was assessed by telephone recall on three occasions over approximately 2 months. After completing the three 24-hour recalls, each calibration subject was sent a second questionnaire; 1,606 persons completed three recalls and a second questionnaire (127 to 267 per ethnic-sex group). This report describes correlation coefficients and calibration slopes for the relation between the 24 hour recalls and second questionnaire values for a selected set of macro- and micronutrients, as absolute intakes, nutrient densities, and calorie-adjusted nutrients. In all subgroups, estimates of the correlation between the questionnaire and 24-hour recalls were greater after energy adjustment (average correlations ranged from 0.57-0.74 for nutrient densities and from 0.55-0.74 for calorie-adjusted nutrients) than when absolute nutrient values were used (average range 0.26-0.57). For absolute nutrient intakes, the correlations were greatest for Whites, somewhat lower for Japanese-Americans and Latinos, and lowest for African-Americans. After energy adjustment, the difference between subgroups were diminished, and the correlations were generally highly satisfactory. PMID- 10695596 TI - Tea consumption and risk of bladder and kidney cancers in a population-based case control study. AB - Recent epidemiologic studies have suggested that tea may be protective against cancers of the urinary tract. The authors examined the association between usual adult tea consumption and risk of bladder and kidney cancers in a population based case-control study that included 1,452 bladder cancer cases, 406 kidney cancer cases, and 2,434 controls. For bladder cancer, the age- and sex-adjusted odds ratios (OR) (95% confidence intervals (CI)) referent to nonusers of tea were 0.9 (0.7, 1.1) for <1.0 cup/day, 1.0 (0.8, 1.2) for 1.0-2.6 cups/day, and 0.9 (0.7, 1.1) for >2.6 cups/day (cutpoints for users based on the tertile distribution among controls). When more extreme cutpoints were used, persons who consumed >5 cups/day (>90th percentile) had a suggestive decreased risk (OR = 0.7; 95% CI 0.5, 1.0), but there was no evidence of a dose-response relation. In analyses stratified by median total beverage intake (2.6 liters/day), there was an inverse association with tea use among persons who consumed less than the median (OR = 0.5; 95% CI 0.3, 0.8) but no association for persons who consumed at or above the median. In contrast, for kidney cancer, there was no association with tea use. Adjustment for site-specific risk factors did not alter these results. This study offers only minimal support for an inverse association between tea consumption and bladder or kidney cancer risk. PMID- 10695597 TI - Occupational lung cancer risk for men in Germany: results from a pooled case control study. AB - Occupational exposures such as crystalline silica, diesel engine exhaust, polycyclic aromatic hydrocarbons, and man-made mineral fibers are strongly suspected to increase lung cancer risk. Two case-control studies in Germany conducted between 1988 and 1996 were pooled for a joint analysis. A total of 3,498 male cases and 3,541 male population controls, frequency matched for age and region, were included in the study. The lifelong history of all jobs and industries was coded and occupational exposures were evaluated by expert rating. Odds ratios, crude and adjusted for smoking and asbestos exposure, were calculated by conditional logistic regression. Job-related evaluation showed a statistically significant increased odds ratio adjusted for smoking among farmers; forestry workers, fishermen, and livestock workers; miners and quarrymen; chemical processors; cabinet makers and related wood workers; metal producers and processors; bricklayers and carpenters; road construction workers, pipelayers and well diggers; plasterers, insulators, and upholsterers; painters and lacquerers; stationary engine and heavy equipment operators; transport workers and freight handlers; and service workers. With regard to specific occupational exposures, elevated odds ratios (OR) (95% confidence intervals (CI)) for lung cancer risk adjusted for smoking and asbestos exposure were observed for man-made mineral fibers (OR = 1.48, 95% CI 1.17, 1.88); crystalline silica (OR = 1.41, 95% CI 1.22, 1.62); diesel engine exhaust (OR = 1.43, 95% CI 1.23, 1.67); and polycyclic aromatic hydrocarbons (OR = 1.53, 95% CI 1.14, 2.04). The risk of asbestos exposure, adjusted for smoking was also increased (OR = 1.41, 95% CI 1.24, 1.60). PMID- 10695598 TI - Risk of breast cancer in relation to the use of injectable progestogen contraceptives and combined estrogen/progestogen contraceptives. AB - Recent studies have suggested that progestogen-only contraceptives and combined estrogen/progestogen oral contraceptives (COCs) may increase the risk of breast cancer among women less than 35 years of age or among recent users. The authors conducted a case-control study, in which cases of breast cancer (n = 484) [corrected] and controls (n = 1,625) hospitalized for conditions unrelated to contraceptive use were interviewed from 1994 to 1997 in hospitals in greater Cape Town, South Africa. The women were aged 20-54 years, resided in a defined area around Cape Town, and were Black or of mixed racial descent. The relative risk for exposure to injectable progestogen contraceptives (IPCs), mostly depot medroxyprogesterone acetate, was 0.9 (95% confidence interval (CI) 0.7, 1.2). There were no consistent associations within categories of age or recency or duration of use. For exposure to COCs, the overall relative risk was 1.2 (95% CI 1.0, 1.5). Among women below age 35 years, the relative risk was 1.7 (95% CI 1.0, 3.0), and it was unrelated to the duration or recency of use. The findings suggest that IPCs do not increase the risk of breast cancer, and that COCs may increase the risk among women below age 35 years, although bias cannot be excluded. PMID- 10695599 TI - Bias in breast cancer analyses due to error in age at menopause. AB - With the use of data from the Nurses' Health Study, the authors confirm the recent argument by Pike et al. (Am J Epidemiol 1998;147:718-21) that various ways of assigning ages at menopause to women with simple hysterectomy produce relative risks for the effect of hormone replacement therapy (HRT) that are biased downward. Different methods of including women with simple hysterectomy consistently produced relative risks for the association between a 1-year increase in duration of HRT use and breast cancer that were underestimates compared with the relative risk obtained when these women were excluded from analysis. The authors used longitudinal data on recalled age at menopause to demonstrate another source of error related to age at menopause: error in recall of age at menopause that increases with time since menopause. PMID- 10695600 TI - Differences in socioeconomic status and survival among white and black men with prostate cancer. AB - After diagnosis with prostate cancer, Black men in the United States have poorer survival than White men, even after controlling for differences in cancer stage. The extent to which these racial survival differences are due to biologic versus non-biologic factors is unclear, and it has been hypothesized that differences associated with socioeconomic status (SES) might account for much of the observed survival difference. The authors examined this hypothesis in a cohort study, using cancer registry and US Census data for White and Black men with incident prostate cancer (n = 23,334) who resided in 1,005 census tracts in the San Francisco Bay Area during 1973-1993. Separate analyses were conducted using two endpoints: death from prostate cancer and death from other causes. For each endpoint, death rate ratios (Blacks vs. Whites) were computed for men diagnosed at ages <65 years and at ages > or =65 years. These data suggest that differences associated with SES do not explain why Black men die from prostate cancer at a higher rate when compared with White men with this condition. However, among men with prostate cancer, SES-associated differences appear to explain almost all of the racial difference in risk of death from other causes.